PMID- 9389971 TI - Origins of ... tinctorial methods in histology. PMID- 9389972 TI - ACP Broad Sheet no 151: September 1997. Investigation of dyslipidaemias. PMID- 9389973 TI - Hepatic histology of patients with HIV infection and chronic hepatitis C treated with interferon. AB - AIMS: To evaluate the histological changes seen in liver biopsies after interferon (IFN) treatment in patients with chronic hepatitis C and human immunodeficiency virus (HIV) infection. METHODS: Twenty four intravenous drug users with chronic hepatitis C were investigated histologically before beginning a 12 month course of IFN treatment and 18 months later. Twelve were HIV positive, without opportunistic or other viral infections (group A), and 12 were HIV negative (group B). RESULTS: According to alanine amino-transferase concentrations, four sustained responders and eight non-responders were found in group A; six sustained responders, five relapsers, and one non-responder were found in group B. HCV RNA became negative in one sustained responder of group A and in the six sustained responders of group B. When histological findings of biopsies performed before therapy and 18 months later were compared, no significant changes in the mean value of Knodell's index and subindices were found in group A, whereas in group B Knodell's index, piecemeal necrosis, and focal hepatocellular necrosis decreased significantly. CONCLUSIONS: In chronic hepatitis C, coinfection with HIV showed a tendency towards a lower response to IFN, although this did not reach statistical significance; however, none of the HIV positive patients developed cirrhosis during the follow up and this should be considered in clinical management of such patients. PMID- 9389974 TI - Gastric mucous neck cell and intestinal goblet cell phenotypes in gastric adenocarcinoma. AB - AIM: To investigate the phenotype of cells comprising diffuse and intestinal-type gastric cancers using monoclonal antibodies to two antigens. One antigen (designated D10) is characteristic of gastric mucous neck cells, cardiac glands, pyloric glands, and Brunner's glands. The second antigen (designated 17NM) is specific to the mucous vacuole of intestinal goblet cells. METHODS: Thirty two gastrectomy specimens with adenocarcinoma were studied. Serial paraffin sections were stained immunohistochemically for D10 and 17NM and histochemically for acid and neutral mucins. The cancers were classified histologically as of either diffuse or intestinal type according to Lauren. RESULTS: Of 15 diffuse-type gastric carcinomas, 11 showed the majority of cancer cells staining for D10 while four were typical signet ring cell cancers staining predominantly for 17NM; five tumours displayed both phenotypes with the two phenotypes segregated in different areas of the tumours. In contrast, of 16 intestinal-type cancers, six expressed 17NM, three D10, five neither antigen, and two expressed both antigens. One indeterminate-type cancer expressed both antigens. The staining of individual cells for D10 and 17NM was mutually exclusive in both diffuse and intestinal types. In contrast to the diffuse cancers, intestinal-type cancers typically expressed either antigen only in occasional small groups of cells and individual cells. CONCLUSIONS: In disease, the gastric stem cell can assume the capacity of the duodenal stem cell for divergent differentiation into either intestinal goblet cells (for example, as in intestinal metaplasia) or Brunner's gland cells (for example, as in pyloric gland/Brunner's gland metaplasia). With neoplastic transformation, this potential for divergent differentiation is maintained and gives rise to diffuse-type cancers that display either the D10 phenotype, the 17NM phenotype, or the clonal expression of both phenotypes. In the more cell cohesive (intestinal-type) tumours, differentiation for antigen expression is poorly developed and more frequently directed towards the intestinal goblet cell phenotype. PMID- 9389975 TI - Detection of parvovirus B19 in macerated fetal tissue using in situ hybridisation. AB - AIMS: To compare the application of a non-radioactive in situ hybridisation (ISH) technique with an immunocytochemical technique for the detection of human parvovirus B19 in formalin fixed, paraffin wax embedded sections of macerated fetal tissue. METHODS: Archived samples of liver, lung or kidney from 19 human fetuses were investigated for parvovirus B19 using a full length digoxigenin labelled DNA probe of 5.5 kb; bound probe was detected using an anti-digoxigenin (alkaline phosphatase) conjugate and visualised using NBT/BCIP. Immunocytochemical detection of parvovirus B19 was performed using a monoclonal mouse antiparvovirus B19 antiserum, with a streptavidin-biotin complex (horse radish peroxidase) method. Cases were selected to provide a range of diagnostic certainty and a range of degrees of macerative degeneration. RESULTS: Parvovirus B19 was found in 15 of 19 cases using the B19 ISH technique compared with 8 of 19 cases using the immunocytochemical technique. The four negative cases were all controls known to be parvovirus B19 free. All ISH positive cases showed excellent staining with low background regardless of extent of maceration and tissue type. In comparison, sections stained by the immunocytochemical method showed considerable non-specific immunoreactivity in many cases, particularly with severe maceration. Kidney and lung tissues gave the cleanest results. CONCLUSIONS: ISH is more effective than the immunocytochemical technique for the detection of human parvovirus B19 in macerated fetal tissue. The lack of detectable background staining with the ISH technique led to easier interpretation suggesting that this technique should be the method of choice for the investigation of parvovirus B19 in macerated postmortem tissues. PMID- 9389976 TI - Effects of oestrogen on the extracellular matrix in the endometrium of postmenopausal women. AB - AIM: To obtain insight into the effects of oestrogen on extracellular matrix (ECM) in the postmenopausal endometrium. METHODS: The distribution of the components of the ECM, including collagen types I, III, IV, and VI, and laminin, was investigated in the human postmenopausal endometrium by an indirect immunofluorescence method with specific monoclonal antibodies and a polyclonal antibody. Collagens were also extracted from the endometrial tissues of postmenopausal women who had or had not been treated with oestrogen for three weeks. RESULTS: Immunohistochemical studies demonstrated that type I collagen was the predominant interstitial collagen, and that types III and VI collagens were absent or very sparsely distributed in the stroma of the postmenopausal endometrium. However, types I, III, and VI collagens were diffusely localised in the stroma of the postmenopausal endometrium after administration of oestrogen. Even though type IV collagen was not seen in the basement membrane of the endometrial glands in the endometrium of postmenopausal women in the absence of oestrogen treatment, both type IV collagen and laminin were localised exclusively in the basement membrane of the endometrial glands in the postmenopausal endometrium after three weeks of oestrogen treatment. The level of type III collagen relative to that of type I collagen was significantly increased (p < 0.01) in the endometrium of oestrogen treated postmenopausal women compared with non-treated postmenopausal women. CONCLUSIONS: Conjugated equine oestrogen might induce changes in the distribution of components and in the composition of the ECM in the endometrium of postmenopausal women. PMID- 9389977 TI - Differential patterns of osteoblast dysfunction in trabecular bone in patients with established osteoporosis. AB - AIMS: To analyse osteoblast function in 153 cases of established osteoporosis as previous work has indicated that osteoporosis is a heterogeneous condition characterised by different patterns of osteoclast and osteoblast dysfunction. METHODS: Histomorphometric data from 153 cases with established osteoporosis was used to analyse osteoblast function, using the following parameters: osteoblast number was assessed using the ratio of osteoblast surface to bone surface (ObS:BS); the percentage of active osteoblasts was assessed by using mineralising surface as a proportion of osteoid surface (sLS + dLS/OS); and the efficiency of active osteoblasts was assessed using the ratio of double to total labelled surface (dLS:tLS). The values of each parameter were standardised using age and sex matched control data and a three dimensional matrix was used to identify groups of patients with similar patterns of altered function. RESULTS: The largest group (60 cases) showed a reduction in all three parameters, while a small group (9 cases) had normal osteoblast function. However, one group showed reduction in osteoblast number only (23 cases), while another group showed a normal number of osteoblasts but both reduced percentage and efficiency of activity (14 cases). The results also suggest that efficiency of activity falls first and that this eventually leads to exit from the active pool. CONCLUSIONS: These results demonstrate the presence of heterogeneity of osteoblast dysfunction in osteoporosis, indicating that the disease is caused by interference at a variety of target sites along the pathway of osteoblast proliferation, differentiation, and activation. Greater understanding of this pathway and of the variety of alterations in the pathway that can occur in osteoporosis may allow more focused therapy for different patient groups identified on the basis of histomorphometric analysis. PMID- 9389978 TI - Haemophilus influenzae: an underrated cause of vulvovaginitis in young girls. AB - AIMS: To establish the common pathogens associated with infective vulvovaginitis in young girls in the local population and to determine current management of this condition. METHODS: A prospective laboratory based survey was carried out over 19 months. A questionnaire was then sent to local general practitioners and hospital doctors. RESULTS: One hundred and six swabs were received during the study period of which 43 (40.5%) yielded organisms recognised as causes of vulvovaginitis. The most common pathogen was group A beta haemolytic streptococcus (19), with Haemophilus influenzae the second most common (11). Candida was isolated on nine occasions. The users' questionnaire had an overall response rate of 52%. Forty one per cent of respondents nominated candida as the most common cause of this condition. Forty six per cent were aware that beta haemolytic streptococci caused juvenile vulvovaginitis, but only four (3.6%) knew that H influenzae was a possible pathogen. The most popular agent for empirical treatment of vulvovaginitis was topical clotrimazole cream, although 24 respondents (22%) prescribed antibiotics that are active against both group A beta haemolytic streptococci and H influenzae. CONCLUSIONS: Although H influenzae is the second most common infective cause of juvenile vulvovaginitis in the local population, most doctors managing these patients were unaware of its importance and may not be prescribing appropriate empirical treatment. PMID- 9389979 TI - Helicobacter pylori does not release cysteamine into gastric juice. AB - AIM: To determine whether Helicobacter pylori releases cysteamine into gastric juice as cysteamine is known to be ulcerogenic. METHODS: Samples of fasting gastric juice were collected from 22 individuals (four women); 10 subjects were H pylori negative. The presence of infection was confirmed by examination and culture of gastric biopsies. Cysteamine in gastric juice was measured by reversed phase gradient high performance liquid chromatography with a detection limit of 10 mumol/l. RESULTS: Cysteamine was not detected in any of the gastric juice samples or in extracts of cultured H pylori. CONCLUSIONS: If H pylori produces cysteamine then the amounts produced are insignificant and are unlikely to explain the association between H pylori infection and the development of duodenal ulcer disease. PMID- 9389980 TI - Juvenile polyposis of the stomach: clinicopathological features and its malignant potential. AB - AIMS: To clarify a clinical entity of juvenile polyposis of the stomach compared with generalised juvenile gastrointestinal polyposis. METHODS: The clinicopathological features of juvenile polyposis dominantly involving the stomach at initial presentation were reviewed in 12 patients (three new patients and nine from the literature). These were compared with 29 cases of generalised juvenile gastrointestinal polyposis. RESULTS: There were three men and nine women with juvenile polyposis of the stomach, aged 10-63 years. Hypoproteinaemia was present in nine patients, anaemia in seven, and a family history of intestinal polyposis in seven. No patient presented with a congenital abnormality. During the observation period, two patients developed colonic juvenile polyps. Gastric polyps invariably affected the antrum and extended to the fundus, eventually becoming more numerous, larger, and more pedunculated. Ten patients required gastrectomy for associated malignancy or uncontrolled protein losing gastropathy. Histological examinations of the resected specimens demonstrated neoplastic tissue arising from juvenile polyps in four of the 12 patients. Atypism in these mixed polyps varied from adenoma to well or moderately differentiated adenocarcinoma. CONCLUSIONS: Juvenile polyposis of the stomach has malignant potential, and may be a separate entity from generalised juvenile gastrointestinal polyposis. PMID- 9389981 TI - Diagnostic distance of high grade prostatic intraepithelial neoplasia from normal prostate and adenocarcinoma. AB - OBJECTIVE: To develop a distance measure based methodology to support the morphological evaluation of high grade prostatic intraepithelial neoplasia (PIN), a direct precursor of prostate cancer. METHODS: Eight morphological and cellular features were analysed in 20 cases of high grade PIN found in radical prostatectomy specimens from patients with adenocarcinoma. The diagnostic distance was evaluated to measure the extent to which the feature outcomes of the individual high grade PIN cases differed from the expected outcome profile of normal prostate, low and high grade PIN, and cribriform and large acinar adenocarcinoma. The belief value for high grade PIN was evaluated with a Bayesian belief network (BBN). RESULTS: Complete separation existed between the cumulative absolute diagnostic distances of these 20 cases from the prototype feature outcomes of high grade PIN and normal prostate the values for which were < or = 3 (range 0 to 3) and > or = 9 (range 9 to 15), respectively. The distances from low grade PIN (range 3 to 9), cribriform adenocarcinoma (range 2 to 8), and large acinar adenocarcinoma (range 5 to 10) were intermediate and showed overlap in their distribution. When taking into consideration whether the severity of feature changes was increasing or decreasing in comparison with the category prototype outcomes, the cumulative directional diagnostic distances from high grade PIN ranged from -3 to +3. Positive distance values were seen relative to low grade PIN (range +3 to +9) and relative to normal prostate (range +9 to +15). Negative values were found relative to cribriform adenocarcinoma (range -8 to +2). The distance values from large acinar adenocarcinoma ranged from -2 to +4 and partly overlapped with those from the high grade PIN category. A bivariate scattergram derived from both diagnostic distance measures showed excellent separation between the groups' distances. BBN analysis confirmed the morphology based diagnosis. The distance evaluation resulted in 18 cases whose belief value for high grade PIN ranged from 0.60 to 0.87. In the remaining two cases the results of the BBN analysis showed a belief value of 0.50 and 0.57 for low grade PIN and of 0.49 and 0.38 for high grade PIN, respectively. CONCLUSIONS: Distance measure based methodology represents a useful diagnostic decision support tool for the accurate evaluation of high grade PIN. PMID- 9389982 TI - Local INR correction: justification for a simplified approach. AB - AIMS: Errors in reporting International Normalised Ratios (INR) may be corrected by assignment of a System International Sensitivity Index (System ISI). This 57 centre study tests the validity of several procedures for INR correction. METHODS: Prothrombin times of eight lyophilised coumarin calibrants, a lyophilised normal pool calibrant, and eight frozen coumarin plasmas were determined at each centre. The calibrants were calibrated using international reference preparations. The eight frozen coumarin plasmas were calibrated in a four centre international exercise. The relations tested were: (a) the logarithm of local prothrombin time against the logarithm of reference prothrombin time; (b) reference INR against local prothrombin time; and (c) logarithm of reference INR against logarithm of local prothrombin time. These methods were analysed by both linear and orthogonal regression. RESULTS: All system groups required correction, the mean percentage deviation of the uncorrected data from the calibrated values was 19.0%. There was also considerable variation in INR, with the coefficient of variance (CV) ranging from 11.30 to 17.29%. Correction of INR was possible with all methods (CV reduced to < 7%). However, only when a plot of the logarithm of local prothrombin time against the logarithm of reference prothrombin time was fitted by orthogonal regression, or a plot of logarithm of reference INR against logarithm of local prothrombin time was fitted by either type of regression analysis, did the best fit line through the calibrant plasmas also pass close to the local mean normal prothrombin time. CONCLUSIONS: While INR correction may be achieved by all the above methods, that relating log reference INR to log local prothrombin time by linear regression analysis is the simplest to perform. PMID- 9389983 TI - Screening for bacterial vaginosis: a novel application of artificial nose technology. AB - The AromaScan system was used to analyse vaginal swabs from 68 women attending a genitourinary clinic. Using clinical criteria, subjects were assessed for bacterial vaginosis. After training the AromaScan system to recognise patterns generated from four patients with and four patients without bacterial vaginosis, 16 of the 17 (94%) remaining subjects were correctly identified as having the condition. The positive predictive value of the test was 61.5%. These results indicate that the AromaScan technology may be of value as a screening test for bacterial vaginosis. PMID- 9389984 TI - Origin of hepatocellular carcinoma recurring after allotransplantation revealed by microsatellite analysis. AB - A hepatocellular carcinoma was resected from a liver allotransplant after the patient's original organ had been removed because of a liver carcinoma. DNA analysis was performed to explore the origin of the carcinoma cells. DNA extracted from the carcinoma tissue, from the carcinoma free liver tissue, and from other cells of the recipient underwent polymerase chain reaction amplification for seven microsatellite systems and the X-Y amelogenin system. The allelic pattern from the carcinoma tissue was identical with that from the patient and differed from the DNA profile of the liver tissue. The result confirmed the assumption that the carcinoma tissue had originated from the patient and not from the donor. PMID- 9389985 TI - Lymphoepithelial cyst of the pancreas. AB - A rare case of lymphoepithelial cyst of the pancreas is reported. Microscopically the cyst content consisted of keratinous material and the walls were lined by mature squamous epithelium surrounded by dense lymphoid tissue. Immunohistochemistry showed diffuse reactivity for CD20 and CD3 in the lymphoid tissue and uniform positivity for cytokeratins in the squamous epithelium. Although the histogenesis of lymphoepithelial cysts of the pancreas is not understood, awareness of this lesion is helpful in differentiating it from other pancreatic cystic lesions. PMID- 9389986 TI - Fatal Yersinia enterocolitica transfusion reaction. PMID- 9389988 TI - Oil immersion magnification without the oil. PMID- 9389987 TI - Alcohol estimation at necroscopy: epidemiology, economics and the elderly. PMID- 9389989 TI - Structure and function of somatostatin receptors in growth hormone control. AB - This new generation of SRIF analogs offer exciting opportunities to improve hormone hypersecretion in patients with GH- and TSH-secreting pituitary adenomas and possibly even in patients harboring prolactinomas and non-functioning tumors. Future development of novel analogs with improved affinity for both SSTR2 and SSTR5 may have even greater potency to suppress pituitary hormone hypersecretion and block adenoma growth. PMID- 9389990 TI - Acromegaly: the significance of serum total and free IGF-I and IGF-binding protein-3 in diagnosis. AB - We have studied the physiological and clinical relevance of measurements of serum total and free IGF-I and IGF-binding protein-3 (IGFBP-3) in 57 previously untreated patients with active acromegaly (32 males, 25 females; mean age 47 years) as compared with sex- and age-matched normal healthy controls. Serum total and free IGF-I, but not IGFBP-3, are suitable biochemical parameters for screening for acromegaly. In acromegalics, the mean 24 h serum GH, total IGF-I and IGFBP-3 levels tend to decrease with age. However, in our series of patients, mean 24 h serum GH levels, IGFBP-3, total and free IGF-I do not correlate with disease activity in acromegaly. PMID- 9389991 TI - Growth hormone, insulin-like growth factor-I and its binding proteins in the follow-up of acromegaly. AB - Elevated growth hormone is a cardinal feature of acromegaly from the biological view point. Growth hormone stimulates IGF-I secretion and that of its major binding protein IGFBP-3. In these circumstances, where hyperinsulinaemia is present, IGFBP-1 levels, which are inversely related to insulin, are suppressed. Failure of suppression of growth hormone after oral glucose (> 2 mU/l (1 microgram/l) is the cardinal biochemical feature of acromegaly. IGF-I values at diagnosis are almost invariably raised. There is some overlap in the value of basal IGFBP-3 between normal subjects and acromegalics. For monitoring purposes, growth hormone values, either basal or during the day are useful. There is overlap in the values of IGF-I and IGFBP-3 between normal subjects and patients on treatment. Prognosis in acromegaly is determined by persistent elevation of growth hormone levels above 5 mU/l (2.5 micrograms/ l). More data are required for the prognostic use of IGF-I. PMID- 9389992 TI - New developments in the management of acromegaly. Should we achieve absolute biochemical cure? PMID- 9389993 TI - Is growth hormone bad for your heart? Cardiovascular impact of GH deficiency and of acromegaly. AB - At present, there is growing evidence implicating GH and/or IGF-I in the intricate cascade of events connected with the regulation of heart development and hypertrophy. Moreover, GH excess and/or deficiency have been shown to include in their advanced clinical manifestations almost always an impaired cardiac function, which may reduce life expectancy. This finding is related both to a primitive impairment of heart structure and function and to metabolic changes such as hyperlipidemia, increase of body fat and premature atherosclerosis. Patients with childhood or adulthood-onset GH deficiency have a reduced left ventricular mass and ejection fraction and the indexes of left ventricular systolic function remain markedly depressed during exercise. Conversely, in acromegaly the cardiac enlargement, which is disproportionate to the increase in size of other internal body organs, has been a rather uniform finding. The severity of the acromegalic cardiomyopathy was reported to be correlated better with the disease duration than with circulating GH and/or IGF-I levels. Myocardial hypertrophy with interstitial fibrosis, lymphomononuclear infiltration and areas of monocyte necrosis often results in concentric hypertrophy of both ventricles. The treatment of GH deficiency and excess improved cardiac function. Interestingly, based on the evidence that GH increases cardiac mass, recombinant GH was administered to patients with idiopathic dilated cardiomyopathy. It increased the myocardial mass and reduced the size of the left ventricular chamber, resulting in improvement of hemodynamics, myocardial energy metabolism and clinical status. These promising results open new perspectives for the use of GH in heart failure. PMID- 9389994 TI - Acromegalic arthropathy and sleep apnea. PMID- 9389995 TI - Acromegaly and its treatment. PMID- 9389996 TI - Evidence supporting surgery as treatment of choice for acromegaly. AB - Within a period of fourteen years 531 operations for growth hormone (GH) secreting adenomas were carried out. In this consecutive series 73% of the 396 patients who underwent primary transsphenoidal surgery achieved basal GH levels below 5 micrograms/l, and 58% also had an adequate suppression following an oral glucose tolerance test (OGTT). Slightly less favourable results were found in patients requiring surgery following an initial therapy. However, 41% of 121 such patients, who had either been operated upon previously or who had received external or internal irradiation, nevertheless achieved basal GH levels below 5 micrograms/l after the surgical reintervention. Normal suppression of serum GH during an OGTT was observed in 23% of these patients. The overall complication rate was low and tumour recurrences were very rare. To facilitate easier tumour removal, octreotide was preoperatively administered in 53 patients undergoing primary surgery of large adenomas. Recurrences were documented in a few exceptional cases. These data support our previous experience that once a normal suppression of growth hormone has been documented following surgery of pituitary adenomas, the long-term outcome is favourable. PMID- 9389997 TI - Evidence for the effectiveness of radiotherapy in the treatment of acromegaly. PMID- 9389998 TI - Evidence for dopamine agonists in the treatment of acromegaly. PMID- 9389999 TI - Evidence for octreotide subcutaneously in the treatment of acromegaly. PMID- 9390000 TI - Gametic imprinting in development and disease. PMID- 9390001 TI - Extracellular calmodulin-binding proteins in body fluids of animals. AB - The extracellular calmodulin-binding proteins (CaMBPs) were investigated in body fluids of animals by using the biotinylated calmodulin gel overlay method. Four major CaMBPs with molecular masses of 24, 31.5, 44/45 and 94 kDa were detected in serum, two of 24 and 63 kDa in bovine milk and three of 14, 24 and 52 kDa in human saliva. It suggested that extracellular CaMBPs exist commonly in body fluids of animals, and this result may provide a new clue for understanding the role of extracellular calmodulin. PMID- 9390002 TI - Effect of GH administration on GH and IGF-I receptors in porcine skeletal muscle and liver in relation to plasma GH-binding protein. AB - The present study was undertaken to determine the effect of GH administration on GH and IGF-I receptors in skeletal muscle compared with liver in growing pigs. Plasma IGF-I and GH-binding protein (GHBP) levels were also determined. Twelve Large White pigs (castrated males) were treated daily with 100 micrograms pituitary porcine GH (pGH) per kg body weight or vehicle for 41 days intramuscularly. Relative to controls, pGH administration increased plasma IGF-I concentrations by 3.3-fold. Administration of pGH had no effect on plasma GHBP levels. In liver, 125I-labelled bovine GH (bGH)-specific binding (P < 0.05) and GH receptor (GHR) mRNA levels (P < 0.05) were higher in pGH-treated than in control pigs. In longissimus dorsi (LD), 125I-labelled bGH specific binding did not differ significantly between the two groups while GHR mRNA levels (P < 0.05) were lower in pGH-treated than in control pigs. Administration of pGH had no effect on 125I-labelled bGH-specific binding and GHR mRNA levels in trapezius (TR). 125I-Labelled IGF-I-specific binding in liver was unaffected by pGH administration. Similarly, in liver, LD and TR, IGF-I receptor mRNA levels were not different between pGH-treated and control animals. It can be concluded that (1) GH binding and IGF-I receptor mRNA are not affected by GH in skeletal muscle, (2) GH influences GHR in a tissue-specific manner and (3) hepatic GHR and GHBP levels are not co-regulated. PMID- 9390003 TI - Ontogeny of inhibin secretion in the rat testis: secretion of inhibin-related proteins from fetal Leydig cells and of bioactive inhibin from Sertoli cells. AB - The ontogeny of inhibin secretion in the testis of rats was investigated. Testicular localization, content of immunoactive and bioactive inhibin and its molecular size in fetal and neonatal rats (from 16 days of gestation to 5 days of age) were determined. Strong immunostaining with an antiserum against a polypeptide of porcine inhibin alpha-subunit was noted in testicular interstitial cells from 16 days of gestation. Co-localization of inhibin alpha-subunit and 3 beta-hydroxysteroid dehydrogenase (3 beta HSD) was observed in the interstitial cells until 2 days of age. Immunoreactive inhibin alpha-subunit in the interstitial tissue had disappeared by 5 days of age, although 3 beta HSD positive cells were still detected. Weak immunostaining for the inhibin alpha subunit was detected in the seminiferous tubules, probably in the cytoplasm of Sertoli cells, from 20 days of gestation onward. No inhibin alpha-subunit immunostaining was observed in germ cells throughout the experimental period. Testicular inhibin was detected at 16 days of gestation (49.5 +/- 6.7 pg per testis) by RIA. Testicular immunoreactive inhibin showed a tendency to increase during fetal life and levels were maintained at a similar value after birth (697.0 +/- 46.9 pg per testis at 5 days of age). Inhibin bioactivity and its molecular size in testicular homogenate was examined at 17 days of gestation and 0 and 5 days of age. Although no bioactivity was detected at 17 days of gestation, bioactivity was noted at 0 and 5 days of age (177.7 and 1303.9 pg per testis respectively). Immunoblot analysis with antiserum against inhibin alpha subunit revealed only approximately 40 kDa molecular masses in the testis at 17 days of gestation, probably inhibin-related proteins, but not inhibin. At 0 and 5 days of age, a protein of 30 kDa molecular mass, possibly inhibin, was detected as well as material of approximately 40 kDa molecular mass. FSH in the plasma was first detected at 19 days of gestation (1197.0 ng/l), increased towards birth, and thereafter decreased (4588.5 +/- 572.3 ng/l at 21 days of gestation and 2400.0 +/- 179.6 ng/l at 5 days of age). These results indicate that Leydig cells in fetal and neonatal rats produce inhibin-related substances with no inhibin bioactivity, whereas Sertoli cells begin to produce inhibin during the perinatal period as a possible regulator of FSH secretion. PMID- 9390004 TI - Regulation of the mouse cellular retinoic acid-binding protein-I gene by thyroid hormone and retinoids in transgenic mouse embryos and P19 cells. AB - The regulation of mouse cellular retinoic acid-binding protein-I (CRABP-I) gene expression by the retinoids and thyroid hormones was examined, by using a beta galactosidase (lacZ) reporter gene and a CRABP-I specific antibody, in transgenic mouse embryos and a mouse embryonal carcinoma cell line P19. The CRABP-lacZ reporter gene expression recapitulated the expression pattern of endogenous CRABP I in the developing central nervous system. In mid-gestation mouse embryos the expression of both the transgene and the endogenous protein was elevated under the condition of hypovitaminosis A, suggesting that depletion of retinoic acid (RA) induced CRABP-I expression in embryos. Consistently, this reporter was suppressed by RA in P19 cells. In co-transfection experiments it was demonstrated that the expression of RAR beta, RAR gamma or RXR alpha suppressed this reporter expression. In experiments designed to alter the thyroid hormone status in animals it was demonstrated that both the reporter gene and the endogenous CRABP I expression were reduced by triiodothyronine injection and were elevated in a hypothyroidic condition induced by feeding with iodine-deficient diet supplemented with 6-propyl-2-thiouracil. In co-transfection experiments it was also demonstrated that the expression of T3R beta suppressed the reporter expression in P19 cells. It was concluded that RA had a suppressive effect on CRABP-I gene expression in embryos and P19 cells and the effect could be mediated through RAR beta, RAR gamma or RXR alpha. A role of thyroid hormones in CRABP-I gene expression and vitamin A metabolism in animals is discussed. PMID- 9390005 TI - Ontogenic and nutritional changes in circulating insulin-like growth factor (IGF) I, IGF-II and IGF-binding proteins in growing ewe and ram lambs. AB - The ontogeny of the IGF endocrine system was investigated in 15 young lambs before and after weaning at 62 days of age. Before weaning, plasma IGF-I concentrations were higher in rams than ewes, and plasma concentrations of IGF-II and IGF-binding protein-3 (IGFBP-3) also tended to be higher in rams than in ewes. Feed intake of ewes and rams was restricted after weaning to remove sex differences in feed intake. Plasma concentrations of IGF-I and IGFBP-3 did not differ between rams and ewes at 100 days of age, but plasma IGF-II was higher in rams than in ewes at this time. Since circulating concentrations of GH were higher in rams than in ewes at 100 days of age, this implies that the restricted feed intake blocked the IGF-I and IGFBP-3 responses to GH. We conclude that sex differences in circulating IGF-I and IGFBP-3 concentrations in the growing lamb alter with age, and are not present when nutrition is restricted. PMID- 9390006 TI - Improvement of glucose homeostasis and hepatic insulin resistance in ob/ob mice given oral molybdate. AB - Molybdate (Mo) exerts insulinomimetic effects in vitro. In this study, we evaluated whether Mo can improve glucose homeostasis in genetically obese, insulin-resistant ob/ob mice. Oral administration of Mo (174 mg/kg molybdenum element) for 7 weeks did not affect body weight, but decreased the hyperglycaemia (approximately 20 mM) of obese mice to the levels of lean (L) (+/+) mice, and reduced the hyperinsulinaemia to one-sixth of pretreatment levels. Tolerance to oral glucose was improved: total glucose area was 30% lower in Mo-treated mice than in untreated ob/ob mice (O), while the total insulin area was halved. Hepatic glucokinase (GK) mRNA level and activity were unchanged in O mice compared with L mice, but the mRNA level and activity of L-type pyruvate kinase (L-PK) were increased in O mice by 3.5- and 1.7-fold respectively. Mo treatment increased GK mRNA levels and activity (by approximately 2.2-fold and 61% compared with O values), and had no, or only a mild, effect on the already increased L-PK variables. mRNA levels and activity of the gluconeogenic enzyme, phosphoenolpyruvate carboxykinase (PEPCK) were augmented in O liver (sixfold and by 57% respectively), and these were reduced by Mo treatment. Insulin binding to partially purified receptors from liver was reduced in O mice and restored by Mo treatment. Despite this correction, overall receptor tyrosine kinase activity was not improved in Mo mice. Moreover, the overexpression (by two- to fourfold) of the cytokine tumour necrosis factor alpha (TNF alpha) in white adipose tissue, which may have a determinant role in the insulin resistance of the O mice, was unaffected by Mo. Likewise, overexpression of the ob gene in white adipose tissue was unchanged by Mo. In conclusion, Mo markedly improved glucose homeostasis in the ob/ob mice by an insulin-like action which appeared to be exerted distal to the insulin receptor tyrosine kinase step. The blood glucose-lowering effect of Mo was unrelated to over-expression of the TNF alpha and ob genes in O mice, but resulted at least in part from attenuation of liver insulin resistance by the reversal of pre-translational regulatory defects in these mice. PMID- 9390007 TI - Inhibin, activin and follistatin bind preferentially to the transformed species of alpha 2-macroglobulin. AB - alpha 2-Macroglobulin (alpha 2-M), a major serum glycoprotein, has been implicated as a low-affinity binding protein for inhibin and activin. In serum, alpha 2-M exists as two major species, a native form that is abundant and stable, and a transformed ('fast') species that is rapidly cleared from the circulation via alpha 2-M receptors. In this study inhibin, activin and their major binding protein follistatin were investigated for their ability to bind to the native or transformed species of purified human alpha 2-M. Using native PAGE and size exclusion chromatography, radiolabelled inhibin, activin and follistatin bound to the transformed alpha 2-M. Inhibin and follistatin did not bind significantly to native alpha 2-M, whereas activin was able to bind to the native species but with a lower capacity compared with that to transformed alpha 2-M. Under reducing conditions, binding of these hormones to alpha 2-M was abolished. These findings implicate alpha 2-M as a mechanism whereby inhibin, activin and follistatin may be removed from the circulation through alpha 2-M receptors, but also whereby activin can be maintained in the circulation through its ability to bind to native alpha 2-M. PMID- 9390008 TI - The melanocortin 1, 3, 4 or 5 receptors do not have a binding epitope for ACTH beyond the sequence of alpha-MSH. AB - ACTH(1-39), and several shorter N- and/or C-terminally truncated fragments of ACTH, with and without N-terminal acetylation and/or C-terminal amidation, were tested for binding on a single eukaryotic cell line transiently and independently expressing the melanocortin MC1, MC3, MC4 and MC5 receptors. The results show that none of these MC receptors has specific binding epitopes for the ACTH peptides beyond the amino acid sequence of alpha-MSH, when tested for their ability to compete with 125I-labelled [Nle4,D-Phe7]alpha-MSH and ACTH. The MC3 receptor favours the natural desacetylated N-terminal end of the ACTH peptides, and it has generally more than 10-fold higher affinity for the ACTH peptides than the MC4 receptor. Considering earlier anatomical localisation data, together with the present data, we suggest that the MC3 receptor is the most likely candidate of the MC receptors to mediate the short-loop negative feedback release of corticotrophin-releasing factor (CRF) caused by ACTH/MSH peptides. PMID- 9390009 TI - Treatment effects of intranasal growth hormone releasing peptide-2 in children with short stature. AB - Growth hormone-releasing peptide (GHRP)-2 is a synthetic six amino acid peptide that is a potent GH secretagogue. Although it shares no structural homology with GH-releasing hormone, in clinical studies its actions on the pituitary release of GH are similar. It is effective when administered orally and intranasally. For children with GH deficiency, such noninvasive treatments are most desirable and in need of development. Fifteen children with short stature participated in this study. All of the children had a height < 2 S.D. below mean for age, poor height velocity, delayed bone age, and low serum concentrations of IGF-1. These children had been tested with standard GH secretagogues, e.g. arginine, insulin, and L dopa. Fifty percent of the children were GH deficient, the remainder had idiopathic short stature. The children received testing with GHRH and GHRP-2 as an acute i.v. bolus of 1 microgram/kg; all children in this study demonstrated a GH response > 20 micrograms/l. Each child in this study also demonstrated a GH response > 10 micrograms/l in response to intranasal GHRP-2, in the dose range of 5-20 micrograms/kg. The children were administered intranasal GHRP-2, 5-15 micrograms/kg, twice a day for 3 months, then three times a day. Fifteen children participated in the study for 6 months; six of the children have participated for 18-24 months. Height velocity, serum IGF-1, IGF-binding protein 3 (IGFBP-3) and GH-binding protein (GHBP) concentrations, and GH responses to GHRP-2 by i.v. bolus and intranasal spray were examined during treatment. Height velocity increased from 3.7 +/- 0.2 cm/year to 6.1 +/- 0.3 cm/year at 6 months, 6.0 +/- 0.4 cm/year at 18-24 months. There were no significant changes in IGF-1 or IGF PB3 concentrations, or in acute GH responses to i.v. or intranasal GHRP-2. GHBP concentrations rose significantly, from 439 +/- 63 pmol/l to 688 +/- 48 pmol/l. In this study, intranasal GHRP-2 administration was well tolerated, and produced a modest but significant increase in growth velocity. PMID- 9390010 TI - Effects of clodronate-containing liposomes on testicular macrophages and Leydig cells in vitro. AB - We undertook the present studies to determine if clodronate-containing liposomes have direct effects on Leydig cells. Macrophages and Leydig cells were isolated and maintained separately in culture. Following treatment with clodronate containing liposomes, macrophages were killed in a dose-response fashion over a range of 5-200 microliters liposomes. By comparison, a 500 microliters dose was required to kill Leydig cells, but this was not dependent upon clodronate since liposomes containing buffer elicited an identical response. The concentration of testosterone in medium from Leydig cells treated with clodronate-containing liposomes was significantly reduced compared with untreated cells. However, we subsequently found that liposomes can adsorb testosterone. Therefore, testosterone production was determined at various times following removal of liposomes from Leydig cells, thereby circumventing this complication. It was found that testosterone production was not altered by liposomes under these conditions. Finally, free clodronate had no effect on testosterone production, even at doses representing the amount present within the 500 microliters dose of liposomes. In summary, clodronate-containing liposomes killed testicular macrophages at a far smaller dose than required to kill Leydig cells. Most importantly, neither liposomes no free clodronate had a direct effect on testosterone production. Thus, clodronate-containing liposomes represent a valuable tool to study Leydig cell-macrophage interactions. PMID- 9390011 TI - Insulin-like growth factor-I, actin, and myosin heavy chain messenger RNAs in skeletal muscle after an injection of growth hormone in subjects over 60 years old. AB - Growth hormone (GH) increases the amount of insulin-like growth factor-I (IGF-I) mRNA in rat skeletal muscle, but this effect has not been demonstrated in human muscle. An autocrine effect of IGF-I produced in muscle may be an important determinant of the increased muscle mass associated with GH therapy. Thus, we examined IGF-I mRNA abundance in skeletal muscle biopsy samples taken 10 h after a subcutaneous injection of GH (0.03 mg/kg, n = 6) or placebo (normal saline, n = 5) in men and women over 60 years of age. Relative tissue concentrations of IGF-I mRNA were evaluated with a competitive reverse transcriptase-polymerase chain reaction assay. Mean plasma IGF-I concentrations rose steadily after the GH injection, and were 74% higher in the GH group than in the control group at the time of the muscle biopsies. There was no consistent difference between the GH and control groups in muscle IGF-I mRNA abundance when expressed in relation to total RNA or polyadenylated RNA. However, one GH-treated subject had three times more IGF-I mRNA, relative to polyadenylated RNA, than the average control subject. There was no effect of GH on levels of mRNAs encoding the most abundant myofibrillar proteins, actin and myosin heavy chain. These data do not support the hypothesis that increased IGF-I mRNA abundance in skeletal muscle is required for the anabolic effect of GH in people over 60 years of age. PMID- 9390012 TI - Opioidergic regulation of prolactin secretion during pregnancy: role of ovarian hormones. AB - We have recently demonstrated the existence of a neuromodulatory regulation of prolactin secretion by the opioid system showing a paradoxical opioid-induced prolactin suppression at the end of pregnancy. The aim of this study was to determine a possible interaction between the opioid system and ovarian hormones on the release of prolactin during pregnancy. Serum prolactin levels measured at 1800 h were significantly higher on days 3 and 6 of pregnancy when compared with the other days of gestation. These increases in serum prolactin were reduced significantly by naloxone (2 mg/kg) administered at 1730 h and by RU-486 (10 mg/kg) administered at 0800 h. The response induced by RU-486 was potentiated by naloxone only on day 3. On days 7, 13 and 16, prolactin secretion was not modified by RU-486 and/or naloxone treatment. In RU-486 pretreated rats, naloxone administration increased serum prolactin levels only on day 16 of pregnancy. Interestingly, progesterone treatment (0.5 mg/rat) on days 13, 14 and 15 of pregnancy prevented the high increase in serum prolactin induced by RU-486 and naloxone on day 16 of pregnancy. The progressive increase and decrease of serum progesterone concentration during pregnancy was not modified by naloxone treatment. The participation of oestrogen in the regulation of prolactin secretion by the opioid system on days 3, 16 and 19 was examined by treating these groups of rats with oestradiol benzoate or tamoxifen citrate. Oestradiol (2 micrograms/rat) significantly increased serum prolactin levels on day 3 and naloxone administration did not modify this increase. No effect was observed after oestradiol (5 micrograms/rat) and naloxone treatment on days 16 and 19 of pregnancy. Oral administration of tamoxifen (500 micrograms/kg) the previous day did not modify the serum prolactin concentration measured at 1800 h in oil treated rats on days 3, 16 and 19 of pregnancy. The antioestrogen completely abolished the naloxone-induced prolactin secretion on day 16 in rats pretreated with RU-486 but no effect was observed on day 19. When tamoxifen was administered on days 14 and 15 of pregnancy, the high serum prolactin levels on day 19 induced by treatment with RU-486 and naloxone were significantly reduced. In conclusion, these results provide an important new insight into the existence of a dual neuromodulatory regulation of prolactin secretion by the opioid system during pregnancy. After a stimulatory action during the first days, there is a change to an inhibitory control at the end of pregnancy, starting around day 16. Moreover, the activation of the inhibitory modulation of the opioid system on prolactin secretion on days 16 and 19 of pregnancy seems to be mediated by changes in the oestrogen and progesterone action. PMID- 9390013 TI - The neuroendocrine control of clock-timed gonadotropin release in the female Syrian hamster: role of serotonin. AB - We hypothesized that rhythms in hypothalamic serotonergic activity were permissive to daily and estrous cycle-related rhythms of LH, FSH and prolactin (PRL). In the Syrian hamster, proestrus (PRO) is characterized by a surge of LH, FSH and PRL; diestrus (DIE) by low LH and FSH and a small surge of PRL, while in photoperiod-induced anestrous (PIA) animals there is a surge of LH and FSH and low PRL. Turnover rates of serotonin (5HT) in four brain areas were determined for the three reproductive states at 2-h intervals. Turnover in the preoptic area and arcuate nuclei did not change, indicating that 5HT projections to these regions probably do not control LH, FSH or PRL release. Serotonin turnover in the median eminence (ME) was elevated at 0600 h in PIA females, at 0600 h, 0800 h, and 1400 h on DIE and at 0600 h and 2200 h on PRO. Since the pattern of 5HT turnover in the ME is different during each of the three reproductive states, 5HT in this area is likely not crucial to the control of LH, FSH and PRL. Turnover of 5HT also did not change in the suprachiasmatic nuclei (SCN) of PRO or PIA animals. However, 5HT turnover rates in the SCN were elevated at 1200 h, 2000 h, and 2400 h on DIE. The correlation of high 5HT turnover in the SCN of DIE but not PRO and PIA animals suggested that elevated serotonergic activity in the SCN is part of the mechanism by which the gonadotropin surge is prevented on DIE. To test this, PRO and DIE hamsters were injected with 5HT receptor ligands. Administration of a 5HT agonist attenuated the PRO surge of LH and blocked the surge of PRL. In contrast, administration of two 5HT antagonists failed to elicit a surge of LH in DIE and phenobarbital-blocked PRO females, an indication that other mechanisms also contribute to inhibition of gonadotropin and PRL surges. PMID- 9390014 TI - Evidence that gonadotropin-releasing hormone (GnRH) functions as a prolactin releasing factor in a teleost fish (Oreochromis mossambicus) and primary structures for three native GnRH molecules. AB - Three forms of gonadotropin-releasing hormone (GnRH) are isolated and identified here by chemical sequence analysis for one species of tilapia, Oreochromis niloticus, and by HPLC elution position for a second species of tilapia, O. mossambicus. Of the three GnRH forms in O. mossambicus, chicken GnRH-II (cGnRH II) and sea bream GnRH (sbGnRH) are present in greater abundance in the brain and pituitary than salmon GnRH (sGnRH). These three native forms of GnRH are shown to stimulate the release of prolactin (PRL) from the rostral pars distalis (RPD) of the pituitary of O. mossambicus in vitro with the following order of potency: cGnRH-II > sGnRH > sbGnRH. In addition, a mammalian GnRH analog stimulated the release of PRL from the pituitary RPD incubated in either iso-osmotic (320 mosmol/l) or hyperosmotic (355 mosmol/l) medium, the latter normally inhibiting PRL release. The response of the pituitary RPD to GnRH was augmented by co incubation with testosterone or 17 beta-estradiol. The effects of GnRH on PRL release appear to be direct effects on PRL cells because the RPD of tilapia contains a nearly homogeneous mass of PRL cells without intermixing of gonadotrophs. Our data suggest that GnRH plays a broad role in fish, depending on the species, by affecting not only gonadotropins and growth hormone, but also PRL. PMID- 9390015 TI - Adaptation of parathyroid function to intravenous 1,25-dihydroxyvitamin D3 or partial parathyroidectomy in normal dogs. AB - Parathyroid function was studied in 14 normal dogs 1 month before and after daily i.v. administration of 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) (eight dogs), or about 50% parathyroidectomy (six dogs), to test the hypothesis that degradation of newly synthesized intact parathyroid hormone (I-PTH) is involved in parathyroid gland adjustment to a modified demand for I-PTH. Parathyroid function was studied through i.v. infusions of Na2EDTA and CaCl2 and measurement of ionized calcium (Ca2+), I-PTH and carboxyl-terminal PTH (C-PTH) at various time points. The C-PTH/I-PTH ratio was used as an index for change in the relative proportion of circulating C-PTH vs I-PTH, 1 month prior to and following each intervention. This ratio was further validated by looking at the HPLC profile of I- and C-PTH in hypo- and hypercalcemia under experimental conditions. Basal Ca2+ was unaltered 1 month after surgery, and was maintained constant in the 1,25 (OH)2D3-treated group by gradually decreasing 1,25-(OH)2D3 doses over time from 0.25 to 0.13 microgram twice daily during the last week of the experimental protocol. In this same group, basal 1,25-(OH)2D3 was increased by 65% (P < 0.0001) and basal I-PTH was decreased by 40% (P < 0.05), while basal C-PTH and the C-PTH/I-PTH ratio remained unchanged. Stimulated and non-suppressible I- and C-PTH followed the same pattern with, this time, an increase of stimulated and non-suppressible C-PTH/I-PTH ratio of 60% (P < 0.05) and 85% (P < 0.05) respectively. There was no change in basal I-PTH, C-PTH, or C-PTH/I-PTH ratio after surgery. However, stimulated I- and C-PTH were decreased by 45% (P < 0.005) and 65% (P < 0.005) respectively, with a 30% (P < 0.005) decrease of stimulated C PTH/I-PTH ratio. There was no change in non-suppressible I-PTH, while non suppressible C-PTH decreased by 55% (P < 0.005), with a 55% (P < 0.05) decrease in non-suppressible C-PTH/I-PTH ratio. The HPLC profiles of I- and C-PTH obtained in hypo- and hypercalcemia disclosed a similar distribution of the immuno reactivity into peaks before and after i.v. administration of 1,25-(OH)2D3 as well as partial parathyroidectomy. This indicated that C-PTH/I-PTH ratio changes were related to different circulating levels of I- and C-PTH rather than to a different composition of I- and C-PTH. These data indicate a shift in the circulating PTH profile toward more PTH carboxyl-terminal fragments after 1 month of i.v. 1,25-(OH)2D3, but toward more intact PTH 1 month after about 50% parathyroidectomy, possibly reflecting adjustments in PTH degradation induced by a modified demand for I-PTH. Although these changes are most likely modulated at the parathyroid gland level, we cannot formally eliminate participation of the hormone's peripheral metabolism. PMID- 9390016 TI - Developmental regulation of preproenkephalin (PENK) gene expression in the adrenal gland of the ovine fetus and newborn lamb: effects of hypoxemia and exogenous cortisol infusion. AB - Development of the fetal adrenal gland is crucial not only for maturation of several fetal organ systems and the initiation of parturition, but also for the development of the fetal response to stress. The enkephalin-related peptides are present in the chromaffin cells of the fetal adrenal medulla and are secreted in response to stress and with sympathetic stimulation. However, changes in expression of preproenkephalin (PENK) with gestation and in response to stress have not been studied in detail. Therefore we examined the developmental pattern of PENK gene expression in the adrenal gland of fetal and newborn lambs, and of adult sheep. We also determined whether levels of PENK mRNA in the fetal adrenal gland changed in response to exogenous glucocorticoids in late gestation, or in response to hypoxemia. Adrenal glands were removed from fetal sheep, lambs and adult sheep at different stages of development for measurement of PENK mRNA. Cortisol was infused (5 micrograms/min) for 12, 24 or 96 h beginning on day 124 129 of gestation. Moderate hypoxemia was induced for 48 h beginning on day 126 130, or at day 134-136 of gestation, by lowering the maternal fractional inspired oxygen. At the end of the treatment periods, the ewes and fetuses were euthanized. Adrenal PENK mRNA were measured by Northern blot analysis. PENK mRNA levels in fetal adrenals were significantly higher (P < 0.05) on days 140-141 of gestation than earlier in pregnancy, and then decreased significantly with the onset of parturition (days 142-146). After cortisol infusion to the fetus for 96 h there was a significant reduction in adrenal PENK mRNA levels. Hypoxemia resulted in a significant increase in PENK mRNA levels in fetuses at day 126-130 of gestation, but not at the later time in pregnancy when endogenous plasma cortisol concentrations were higher. We conclude that there is a decrease in levels of PENK mRNA in the fetal adrenal gland before parturition at the time of the endogenous prepartum rise in plasma cortisol. Hypoxemia led to an elevation of PENK mRNA levels in fetuses at less than 130 days, but after that time, when the basal and stimulated cortisol responses had risen, there was no significant effect of hypoxemia on PENK mRNA. Cortisol infusion to the fetus at this stage of pregnancy resulted in a decrease in adrenal PENK mRNA levels. We suggest that cortisol may play an important role in the regulation of fetal adrenal PENK mRNA levels and enkephalin synthesis by the adrenal gland of the fetal sheep. PMID- 9390017 TI - Effect of thyroid state on lipid peroxidation, antioxidant defences, and susceptibility to oxidative stress in rat tissues. AB - The effects of altered thyroid states on lipid peroxidation, antioxidant capacity, and susceptibility to oxidative stress of rat tissues were examined. Hypothyroidism was induced by administering methimazole in drinking water for 15 days. Hyperthyroidism was elicited by a 10-day treatment of hypothyroid rats with tri-iodothyronine (10 micrograms/100 g body weight). In tissues of hypothyroid rats the lipid peroxidation was not modified, whereas in hyperthyroid rats lipid peroxidation increased in liver and heart but not in skeletal muscle. The glutathione peroxidase activity increased significantly in heart and muscle of hypothyroid rats and in muscle of hyperthyroid rats. The glutathione reductase activity was not modified in tissues of hypothyroid and hyperthyroid rats. In both rat groups the whole antioxidant capacity of tissues decreased, but significantly only in liver and heart. The results obtained studying the response to oxidative stress in vitro indicated that the susceptibility to oxidative challenge was increased in all tissues of hyperthyroid rats and in heart and muscle of hypothyroid animals. These results are explainable in terms of tissue variations in haemoprotein content and/or of antioxidant capacity. Since it has been reported that hypothyroidism offers in vivo protection against free radical damage, we suggest that such an effect could be due to greater effectiveness of cellular defence systems different from antioxidant ones. PMID- 9390018 TI - Estrogen enhances growth hormone receptor expression and growth hormone action in rat osteosarcoma cells and human osteoblast-like cells. AB - Postmenopausal bone loss is primarily due to estrogen deficiency. Recent clinical observation demonstrate that GH increases bone mass in GH deficient patients. The present study investigates whether estrogen regulates GH action and GH receptor expression in osteoblasts. 17 beta-estradiol or GH added to the culture medium as single substances did not influence rat osteosarcoma cell proliferation nor human osteoblast-like (hOB) cell proliferation. However, together they synergistically induced osteoblast proliferation (rat osteosarcoma cells 160.1 +/- 15.5% of control cells; human osteoblast-like cells 159.6 +/- 5.1% of control cells). 17 beta-estradiol stimulated 125I-GH binding and GH receptor (GHR) mRNA levels in rat osteosarcoma cells. The stimulatory effect of estradiol was time dependent, reaching a peak after 8 h of incubation with 17 beta-estradiol (binding 216.9 +/- 27.8% and mRNA 374.6 +/- 30.8% of control). The finding that estradiol stimulated 125I-GH binding was confirmed in human osteoblast-like cells. In these cells, 17 beta-estradiol (10(-12) M) increased 125I-GH binding to 203.8 +/- 3.6% of control levels. We conclude that estrogen stimulates GH activity as well as GH binding and GHR mRNA levels in osteoblasts. These findings indicate that estrogen potentiates the effect of GH at the receptor level. PMID- 9390019 TI - T and B cell development in pituitary deficient insulin-like growth factor-II transgenic dwarf mice. AB - Treatment of mice with IGF-I stimulates T and B cell development. We showed that overexpression of IGF-II in transgenic FVB/N mice only stimulated T cell development. In the present study, we further addressed the in vivo effects of IGF-II in the absence of IGF-I to get more insight into the potential abilities of IGF-II to influence T and B cell development. To this end, we studied lymphocyte development in IGF-II transgenic Snell dwarf mice that are prolactin, GH and thyroid-stimulating hormone deficient and as a consequence show low serum IGF-I levels. We showed that T cell development was stimulated to the same extent as in IGF-II transgenic FVB/N mice. Furthermore, IGF-II increased the number of nucleated bone marrow cells and the number of immature B cells without having an effect on the number of mature B cells in spleen and bone marrow. Our data show that IGF-II has preferential effects on T cell development compared with B development, and that these preferential effects also occur in the absence of measurable IGF-I levels. PMID- 9390021 TI - Who wants book reviews? PMID- 9390020 TI - Inhibition of 11 beta-hydroxysteroid dehydrogenase activity enhances the antiproliferative effect of glucocorticosteroids on MCF-7 and ZR-75-1 breast cancer cells. AB - This in vitro study on MCF-7 and ZR-75-1 breast cancer cells showed that the antiproliferative action of glucocorticosteroids (GCS) on breast cancer cells is weakened by a high oxidative activity of 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD; EC 1.1.1.146): both endogenic as well as synthetic GCS (dexamethasone, prednisolone) were metabolised to hormonally inactive 11-dehydro metabolites. This enzymatic shield protected the breast cancer cells from the antiproliferative action of GCS. Continuous exposure of breast cancer cells to GCS resulted in enhanced 11 beta-HSD activity. The intracellular GCS concentration was further reduced by this feedback and thus the antiproliferative effect was additionally weakened. These mechanisms of GCS deactivation could be influenced by inhibiting 11 beta-HSD with the liquorice compound glycyrrhetinic acid (GLY). In MCF-7 and ZR-75-1 cultures the antiproliferative effect of GCS was significantly increased by GLY. PMID- 9390022 TI - Quantitative microscopy in studies of intrauterine growth retardation. AB - In the past, the detection of fetal damage has tended to be restricted to the naked eye identification of major malformations, with the period of organ maturation being relatively neglected. Increasingly, however, unbiased design based stereology is being used in developmental toxicological studies. In the field of intrauterine growth retardation, such methods are capable of providing new insights into fetal vulnerability during critical periods in organogenesis, with consequences for both post-natal and adult disease. PMID- 9390023 TI - The Heidelberg classification of renal cell tumours. AB - This paper presents the conclusions of a workshop entitled 'Impact of Molecular Genetics on the Classification of Renal Cell Tumours', which was held in Heidelberg in October 1996. The focus on 'renal cell tumours' excludes any discussion of Wilms' tumour and its variants, or of tumours metastatic to the kidneys. The proposed classification subdivides renal cell tumours into benign and malignant parenchymal neoplasms and, where possible, limits each subcategory to the most commonly documented genetic abnormalities. Benign tumours are subclassified into metanephric adenoma and adenofibroma, papillary renal cell adenoma, and renal oncocytoma. Malignant tumours are subclassified into common or conventional renal cell carcinoma; papillary renal cell carcinoma; chromophobe renal cell carcinoma; collecting duct carcinoma, with medullary carcinoma of the kidney; and renal cell carcinoma, unclassified. This classification is based on current genetic knowledge, correlates with recognizable histological findings, and is applicable to routine diagnostic practice. PMID- 9390024 TI - Multiple pathways control cell growth and transformation: overlapping and independent activities of p53 and p21Cip1/WAF1/Sdi1. AB - Many tumour therapies act by inducing a cellular damage response pathway mediated by the tumour suppressor protein p53. Alternative outcomes of p53 induction include apoptosis or transient cell-cycle arrest, both thought to require the transcriptional activity of wild-type p53. Current research highlights the action of a p53-activated gene, p21Cip1/WAF1/Sdi1, which encodes a cyclin-kinase inhibitor important in mediating p53-dependent cell-cycle arrest, while programmed cell death in response to DNA damage requires transcriptionally active p53 but not activation of p21Cip1/WAF1/Sdi1. This review examines the roles of p53 and p21Cip1/WAF1/Sdi1 in controlling cell proliferation, in the light of a new study on expression of p53 and p21Cip1/WAF1/Sdi1 in squamous cell carcinoma of the larynx. PMID- 9390025 TI - Classification strategies for the grading of renal cell carcinomas, based on nuclear morphometry and densitometry. AB - The various grading systems proposed for renal cell carcinomas all suffer from problems related to inter-observer variability. Some of these grading systems are based, either partially or wholly, on morphonuclear criteria, such as nuclear size and shape, anisonucleosis, and chromatin pattern. These criteria can be quantitatively (and thus objectively) evaluated by means of the computer-assisted microscopic analysis of Feulgen-stained nuclei. In the present work, 39 quantitative variables, including two morphometric, 28 chromatin pattern-related, and nine DNA ploidy level-related, were computed for 65 renal cell carcinomas. The actual diagnostic information contributed by each variable was determined by means of multifactorial statistical analysis (discriminant analysis) and two artificial intelligence-related methods of data classification (the decision tree and production rule methods). The results show that quantitative information, as provided by the computer-assisted microscopy of Feulgen-stained nuclei and analysed by means of artificial intelligence-related methods of data classification, contributes significant diagnostic information for the grading of renal cell carcinoma, thus reducing the problem of inter-observer reproducibility. PMID- 9390026 TI - Loss of heterozygosity at chromosomes 8p, 9p, and 14q is associated with stage and grade of non-papillary renal cell carcinomas. AB - In this study, 105 non-papillary renal cell carcinomas (RCCs) have been examined for allelic loss at the chromosome 8p12-21.1, 9p21, and 14q24.2-qter regions, each by two highly polymorphic microsatellites. Loss of heterozygosity (LOH) was detected at both chromosome 8p and 9p in 33 per cent of the cases and at chromosome 14q in 45 per cent of the tumours. A correlation of variables such as size, grade, and stage of tumours with these specific genetic alterations showed that loss of chromosomes 8p and 9p, and especially loss of chromosome 14q regions, is significantly associated with a higher grade of tumour and the combined LOH at these chromosomal sites with advanced tumour stage. These genetic alterations did not show any correlation with the size of non-papillary RCCs. This study suggests that genetic markers at the above-mentioned chromosomal sites can predict the clinical outcome of non-papillary RCCs. PMID- 9390027 TI - p21WAF1/Cip1 expression is associated with cell differentiation but not with p53 mutations in squamous cell carcinomas of the larynx. AB - p21WAF1/Cip1 is a recently identified gene involved in cell cycle regulation through cyclin-CDK-complex inhibition. The expression of this gene in several cell lines seems to be induced by wild-type, but not mutant, p53. p21WAF1/Cip1 expression has been studied at both mRNA and protein levels in a series of 49 normal mucosae and squamous cell carcinomas of the larynx. A significant association was found between mRNA and protein expression in tumours (P < 0.0001). p21WAF1/Cip1 expression was strongly associated with squamous cell differentiation of carcinomas, because six of seven (86 per cent) undifferentiated carcinomas (grade 4) showed very low levels of p21WAF1/Cip1 expression, whereas 41 out of 42 (98 per cent) carcinomas with squamous cell differentiation (grades 1-3) had normal or high levels of p21WAF1/Cip1 expression (P < 0.0001). In addition, p21WAF1/Cip1 expression was topologically related to the squamous differentiation of tumour cells with a distribution similar to that seen in normal squamous epithelium. No correlation was found between p21WAF1/Cip1 expression and the global S-phase of the carcinomas. p53 mutations (exons 5-9) were found in ten carcinomas with p21WAF1/Cip1 expression, but no p53 mutations were detected in three p21WAF1/Cip1-negative tumours. In conclusion, p21WAF1/Cip1 expression is frequently upregulated in squamous cell carcinomas of the larynx and is associated with tumour cell differentiation. p21WAF1/Cip1 expression in these tumours is independent of p53 gene mutations. PMID- 9390028 TI - The association of squamous cell carcinomas of the nasopharynx with Epstein-Barr virus shows geographical variation reminiscent of Burkitt's lymphoma. AB - Nasopharyngeal carcinoma (NPC) is rare in most parts of the world but occurs with high incidence in certain regions, such as South-East Asia. Two major histological types of NPC are recognized, non-keratinizing carcinoma and squamous cell carcinoma. Non-keratinizing NPCs, which include undifferentiated NPC, are invariably associated with Epstein-Barr virus (EBV) infection, regardless of the geographical or ethnic origin of the patients. By contrast, conflicting results have been published concerning a possible association of squamous cell NPC with the virus. To address this question, squamous cell NPCs have been collated from an area where NPC is endemic, Hong Kong, and from two regions where NPC occurs with a lower incidence, Chengdu, PR China, and Birmingham, United Kingdom. In situ hybridization for the detection of the small EBV-encoded nuclear RNAs (EBERs) demonstrated that all 22 cases from Hong Kong were EBV-positive. By contrast, EBV was detectable in 7 of 19 cases from central China, and in 3 of 7 cases from the U.K. Expression of the virus-encoded latent membrane protein 1 (LMP1) was detected in 3 of 32 EBV-positive squamous cell NPCs. These results indicate that the association of squamous cell NPCs with EBV shows geographical variability in a manner which is reminiscent of the situation encountered in Burkitt's lymphoma. This suggests that squamous cell NPCs are a pathogenetically heterogeneous group of tumours distinct from non-keratinizing NPCs. PMID- 9390029 TI - Human ageing impairs injury-induced in vivo expression of tissue inhibitor of matrix metalloproteinases (TIMP)-1 and -2 proteins and mRNA. AB - Proteolysis is an essential component of wound healing but, if uncontrolled, it may lead to degradation of the neo-matrix and a delay in wound repair. Despite numerous reports of impaired wound healing associated with increasing age, the control of proteolysis is completely unknown. Tissue inhibitor of matrix metalloproteinases (TIMP)-1 and -2 inhibit the activity of matrix metalloproteinases and the pattern of regulation of these molecules determines in part the spatial and temporal regulation of proteolytic activity. This study reports on TIMP-1 and -2 protein localization using immunocytochemistry in healing wounds of healthy subjects of different ages from day 1 to 6 months post wounding, and has quantified the mRNA levels for both inhibitors using reverse transcriptase-polymerase chain reaction (RT-PCR). TIMP-1 and TIMP-2 proteins are up-regulated from 24 h post-wounding, with a decrease in staining intensity by day 7 for TIMP-2 and by day 14 for TIMP-1. Steady-state mRNA levels for both TIMPs were significantly greater in normal young skin than in aged skin. In the young, there was a significant increase in mRNA expression for TIMP-1 and -2 by day 3 post-wounding, which decreased by day 14 and had returned to basal levels at day 21. In the wounds of the aged subjects, basal levels were observed for TIMP-1 and -2 at all time-points. These results suggest that intrinsic cutaneous ageing is associated with reduced levels of TIMP mRNA both in normal skin and during acute wound repair. These levels may be instrumental in dermal tissue breakdown in normal skin, retarded wound healing, and the predisposition of the elderly to chronic wound healing states. PMID- 9390030 TI - UV but not gamma-irradiation induces specific transcriptional activity of p53 in primary hepatocytes. AB - The mechanisms are poorly understood by which p53 can stimulate different downstream events, including growth arrest, DNA repair, and apoptosis, after DNA damage. Changes in protein levels do not predict a particular p53 response, but it is possible that differences in functional activities such as transactivation are important. The present report describes the successful use of a specific p53 reporter plasmid transfected into primary murine hepatocytes to evaluate p53 transactivation activity over time after two different genotoxic injuries (gamma irradiation, 15 Gy and UV-c irradiation, 10 J/m2) known to produce p53-dependent growth arrest in this cell type. The results show that UV injury to hepatocytes was followed by a transient increase in transcriptional activation of the reporter plasmid by p53 and that this response preceded changes in p53 protein levels, as assessed by immunocytochemistry. By contrast, gamma-irradiation injury failed to induce detectable changes in either transactivation activity or hepatocyte p53 protein levels. The data show that p53 responses to DNA damage are dependent on both cell and injury type and suggest that in hepatocytes they can be independent of protein concentration and specific transcriptional activity. The results have implications for how particular dysfunctional p53 mutations in carcinogenesis could alter hepatocyte responses to different DNA injuries. PMID- 9390031 TI - In vivo expression of the CTLA4 inhibitory receptor in malignant and reactive cells from human lymphomas. AB - The CTLA4 receptor is a CD28 homologue which induces inhibitory effect on activated T-cells. Peripheral T-cells proliferate spontaneously in CTLA4 deficient mice. These results led to an analysis of CTLA4 expression in human lymphomas (n = 82) including Hodgkin's disease (HD) and non-Hodgkin's lymphomas (NHLs), using immunohistochemistry. CTLA4 was present in neoplastic cells from most (10/11) T-cell malignancies, except for anaplastic and lymphoblastic subtypes (0/4). Malignant B-cells from rare (3/55) B-NHLs (all of follicular subtype) were also CTLA4-positive. Other B-NHLs (52/55) were negative in malignant B-cells and occasionally positive in T-cells. Reactive small lymphocytes, but not Reed-Sternberg cells, from all (12/12) HD cases were strongly CTLA4-positive. The CTLA4 ligands CD80 and CD86 were simultaneously expressed in most CTLA4-negative lymphoma cases. CTLA4 is thus expressed either in the reactive or in the malignant cell populations, depending on the lymphoma subtype. These results provide new insights leading towards therapeutic strategies based either on enhancement of anti-tumour immunity by CTLA4 blockade in reactive lymphocytes or on triggering of a CTLA4-mediated inhibitory pathway in lymphoma cells. PMID- 9390032 TI - 'Revertant' DCIS in human axillary breast carcinoma metastases. AB - Recent experimental evidence obtained in Scid mice has suggested that the metastatic process is in large part epigenetically regulated and undergoes partial reversion once the metastatic process is completed: the metastatic colonies become more engaged in the process of growing in situ than actively metastasizing. Based on this experimental evidence, examples were sought of metastatic human cancers where similar reversion to an in situ growth state was occurring. Review of 200 cases of metastatic human breast cancer revealed a 21 per cent incidence of reversion to a ductal carcinoma in situ (DCIS) growth pattern within axillary nodal metastases. The revertant DCIS areas were characterized by an intact and circumferential basement membrane, as demonstrated by extracellular laminin and type IV collagen immunoreactivity. These revertant DCIS areas could be distinguished from primary DCIS, however, by the absence of surrounding myoepithelial cells in the former, identified in the latter by their positive maspin, S-100, and smooth muscle actin immunoreactivity. The pattern of revertant DCIS, poorly differentiated (comedo) (13 per cent), intermediate (non comedo) (6 per cent), or well-differentiated (non-comedo) (2%), exhibited complete 100 per cent concordance with the primary DCIS pattern. The concordance of histological patterns held true for even the subtypes of DCIS determined by architectural pattern, such as the micropapillary or cribriform subtypes. Nuclear size by digital image analysis and Her-2/neu, p53, and Ki-67 status in the revertant DCIS also exhibited complete concordance with the primary DCIS counterparts. Cases exhibiting a revertant DCIS pattern tended to be ER negative/EGFR-positive and exhibited significant nodal involvement (mean number, 9; mean area, 90 per cent) compared with cases lacking a revertant pattern (mean number, 4; mean area, 15 per cent) (P < 0.01) These findings suggest that reversion of the metastatic phenotype may also be occurring within autochthonous human metastasis. PMID- 9390033 TI - Grb2 overexpression in nuclei and cytoplasm of human breast cells: a histochemical and biochemical study of normal and neoplastic mammary tissue specimens. AB - In 20-30 per cent of human breast cancers, the receptor tyrosine kinases epidermal growth factor receptor (EGFR) and c-erbB2 are overexpressed. This overexpression leads to increased mitogenic signalling and is correlated with poor prognosis. Overexpression of associated adaptor proteins, like Grb2, can also induce upregulation of signalling pathways. In this study, the expression of the Grb2 adaptor protein was determined in both normal human breast tissue and mammary cancers, using immunoblotting experiments and immunostaining on paraffin embedded tissue sections. Both biochemical and immunohistochemical techniques revealed overexpression of Grb2 in all breast cancer specimens. In addition, although Grb2 protein is described as localized in the cytoplasm, it can also be detected in the nucleus, both in normal and in tumour breast tissue. In tumour breast tissue, 58 per cent of Grb2 protein is found in the nucleus, while 37 per cent is detected in the cytoplasm. In normal breast tissue, 22 per cent of Grb2 is found in the nucleus and 70 per cent in the cytoplasm. These findings indicate that in human breast cancer, Grb2 is overexpressed and appears to be predominantly localized in the nucleus. PMID- 9390034 TI - Variation of bcl-2 expression in breast ducts and lobules in relation to plasma progesterone levels: overexpression and absence of variation in fibroadenomas. AB - Some women with benign breast disease eventually develop breast cancer. The mammary gland undergoes tissue remodelling according to hormonal influences, involving a balance between quiescence, proliferation, and mechanisms of cell death. Proliferation and/or apoptotic events could therefore be investigated to help understand the mechanisms of benign lesion formation and identify mastopathies with a poor prognosis. bcl-2 expression was analysed by immunohistochemistry in 75 benign mastopathies. Protein levels were quantitated with an image analyser in various epithelial structures on frozen sections, including adenoses, fibroadenomas, ductal epithelial hyperplasias, cysts, and apparently normal surrounding lobules and ducts. bcl-2 levels were equivalent in apparently normal lobules and ducts, as well as in cysts and ductal hyperplasias. bcl-2 staining was significantly higher in fibroadenomas, known to be of lobular origin [mean = 10.1, quantitative immunochemistry score (QIC) arbitrary units (AU), n = 19], than in normal lobules (mean = 5.1 AU, n = 43, P = 7 x 10(-5). bcl 2 levels in normal lobules and ducts varied according to the menstrual cycle, being higher during the follicular than the luteal phase (P = 1.8 x 10(-2) and P = 1.7 x 10(-2), respectively). This was further supported by a statistical link (P = 5 x 10(-3) between high levels of circulating progesterone and weak bcl-2 staining in lobules and ducts. This progesterone-dependent variation was absent in fibroadenomas. No statistical correlation was found between bcl-2 expression and circulating levels of oestradiol, and follicle-stimulating or luteotrophic hormones. Although these are only preliminary results, they suggest an influence of progesterone on bcl-2 expression which might be lost in fibroadenomas. A hypothesis is proposed concerning the potential involvement of altered regulation of the apoptotic process in the formation of such benign lesions. PMID- 9390035 TI - Expression of the receptor for parathyroid hormone-related protein in normal and malignant breast tissue. AB - Parathyroid hormone-related protein (PTHrP) is the cause of humoral hypercalcaemia of malignancy and interacts with parathyroid hormone (PTH) receptors. Breast cancer cells produce PTHrP in vitro and in vivo. The breast cancer cell line MCF-7, which products PTHrP and expresses PTHrP receptors, proliferates in response to PTHrP. The aim of these studies was to determine the tissue location of PTHrP/PTH receptors (PTHrPR) in primary breast carcinomas and to establish whether they had the potential to respond to PTHrP. The cellular location of mRNA for the PTHrP/PTH receptor was identified using in situ hybridization in primary breast carcinomas and normal breast tissue. Immunohistochemistry for PTHrP was carried out on the same specimens. Tumours were assessed and scored by two observers using the product of intensity of signal and number of positive tumour cells (possible range 0-9). Tumours were also assessed for Ki-67 expression by counting positive nuclei. Non-malignant ductular epithelium expressed mRNA for the PTHrP receptor (mean score 2.6, range 1-4). Breast carcinomas (mean score 4.4, range 0-9) showed variable expression of PTHrP receptor mRNA: eight tumours were negative, 50 had scores similar to normal breast tissue, and 49 had higher scores for the receptor. Levels of expression of the receptor within the primary breast carcinomas were unrelated to immunohistochemical detection of PTHrP or to any standard prognostic factor. There was a significant (P = 0.05) relationship between Ki-67 and PTHrPR expression in individual tumours. The presence of PTHrP and its receptor in normal breast epithelium and breast carcinomas demonstrates that most breast tumours are able to respond to PTHrP. The Ki-67 data suggest that PTHrP is a potential autocrine growth factor in primary breast carcinoma. PMID- 9390036 TI - Expression of apoptosis-suppressing gene bcl-2 in human carotid body tumours. AB - That carotid body tumours have a genetic aetiology is suggested by the familial occurrence of the neoplasm. Environmental influences are also implied by the fact that the tumour is more common in those living at high altitudes. However, the mechanism of development of sporadic tumours occurring at sea level, which account for the majority of cases, remains unknown. It has become increasingly clear that the deregulation of programmed cell death is a critical component in multistep tumourigenesis. Previous studies have demonstrated a high frequency of bcl-2 expression in the tumours arising from cells derived from the neural crest and tumour cell lines of neural origin. This investigation was undertaken to determine whether similar molecular events occur in human carotid body tumours. Western and Northern analysis revealed that the tumours expressed the 26 kD protein and bcl-2 transcripts. Immunoperoxidase staining, using a monoclonal anti bcl-2 antibody, revealed that 11 out of 13 specimens stained positively for bcl 2. These results suggest that the deregulation of programmed cell death may be a critical component in the multistep tumourigenesis of carotid body tumours and that the expression of oncoprotein bcl-2 may contribute to the generation of such tumours. PMID- 9390037 TI - Expression and prognostic value of the CD44 splicing variants v5 and v6 in gastric cancer. AB - In the present study, the expression and prognostic role of the CD44 splicing variants v5 and v6 were immunohistochemically investigated in 418 curatively resected gastric carcinomas. CD44v5 was expressed in 65.3 per cent (n = 273) and CD44v6 in 77.0 per cent (n = 322) of the tumours. Whereas the expression of CD44v5 was correlated with advanced pT categories, with lymph node involvement, and with the presence of blood and lymphatic vessel invasion, such a correlation could not be found for the variant v6. As shown by univariate analysis, patients with CD44v5-positive tumours had a significantly shorter overall survival than patients with CD44v5-negative tumours (P = 0.049). In contrast, expression of CD44v6 had no impact on prognosis (P = 0.574). In a multivariate analysis including the prognostic parameters pT category and pN category, as well as blood and lymphatic vessel invasion, the prognostic impact of CD44v5 expression could not, however, be maintained. Although in the present study the expression of CD44v5 was correlated with a more aggressive tumour type, these data suggest that neither CD44v5 nor CD44v6 can predict survival in patients with gastric cancer, nor is their expression a suitable tool for identifying subgroups of patients who may be at higher risk. PMID- 9390038 TI - New monoclonal antibodies to oestrogen and progesterone receptors effective for paraffin section immunohistochemistry. AB - Assessment of oestrogen and progesterone receptors (ER and PgR) in breast cancer is widely used for the prediction of response to endocrine therapy and as a prognostic marker. Cytosolic assays have been replaced in many centres by immunochemical techniques, which have many advantages including applicability to small samples, simplicity, and cost-effectiveness. This study describes the generation and characterisation of two novel murine monoclonal antibodies recognizing ER and PgR, designated NCL-ER-6F11 and NCL-PGR respectively, which are effective in heat-treated formalin-fixed, paraffin-embedded tissue. The antibodies have been characterized by Western blotting and by immunohistochemistry on normal and pathological breast and other tissues. NCL-ER 6F11 has been shown to compare favourably with a currently available ER antibody. These antibodies may prove of value in the assessment of hormone receptor status in human breast cancer. PMID- 9390039 TI - Full thickness eosinophilia in oesophageal leiomyomatosis and idiopathic eosinophilic oesophagitis. A common allergic inflammatory profile? AB - Oesophageal leiomyomatosis and idiopathic eosinophilic oesophagitis are both extremely rare. The former is a diffuse proliferation of smooth muscle in the muscularis propria, whilst the latter is an idiopathic inflammatory condition, thought to be associated with background atopy and characterized by an infiltrate of eosinophils throughout the full thickness of the oesophagus. However, two recent cases of oesophageal leiomyomatosis showed similar full thickness infiltration of the oesophageal wall by eosinophils and this inflammatory cell infiltrate was investigated in conjunction with one case of idiopathic eosinophilic oesophagitis. All three had a similar allergic profile characterized by CD45RO-positive primed T-lymphocytes, EG2-positive (i.e., activated) eosinophils, and tryptasepositive mast cells, together with gene expression for interleukin 4. Previous descriptions of leiomyomatosis describe an association with systemic mastocytosis and urticaria and the possibility that there is a common underlying allergic component to both disorders is raised. PMID- 9390040 TI - A highly sensitive detection method for immunohistochemistry using biotinylated tyramine. AB - A highly sensitive method for light microscopic immunohistochemistry is described. The increased sensitivity compared with current methodologies is based on the horseradish peroxidase-catalysed deposition of biotinylated tyramine at the sites of immunoreactivity, followed by the detection of the biotin with streptavidin biotin horseradish peroxidase complex. This method is of general applicability in immunohistochemistry and has several important advantages over currently used immunohistochemical detection procedures. The most significant advantage is that several antibodies which to data have been non-reactive, even in antigen-retrieval formalin-fixed, was-embedded sections, now show strong and reproducible immunoreactivity using biotinylated tyramine amplification. In addition, many other antibodies can be used at significantly higher dilutions. PMID- 9390041 TI - Intracytoplasmic sperm injection: offering hope for a term pregnancy and a healthy child? PMID- 9390042 TI - When lifesaving treatment in children is not the answer. PMID- 9390043 TI - The future of preschool vision screening services in Britain. PMID- 9390044 TI - Too soon to market. PMID- 9390045 TI - Opiate detoxification under anaesthesia. PMID- 9390046 TI - UK exempts motor racing from advertising ban. PMID- 9390047 TI - UK bans powerful laser pointers. PMID- 9390048 TI - Oregon reaffirms assisted suicide. PMID- 9390049 TI - Methadone treatment increases in EU. PMID- 9390050 TI - Milk intake and bone mineral acquisition in adolescent girls: randomised, controlled intervention trial. AB - OBJECTIVES: To investigate the effect of milk supplementation on total body bone mineral acquisition in adolescent girls. DESIGN: 18 month, open randomised intervention trial. SUBJECTS: 82 white girls aged 12.2 (SD 0.3) years, recruited from four secondary schools in Sheffield. INTERVENTION: 568 ml (one pint) of whole or reduced fat milk per day for 18 months. MAIN OUTCOME MEASURES: Total body bone mineral content and bone mineral density measured by dual energy x ray absorptiometry. Outcome measures to evaluate mechanism included biochemical markers of bone turnover (osteocalcin, bone alkaline phosphatase, deoxypyridinoline, N-telopeptide of type I collagen), and hormones important to skeletal growth (parathyroid hormone, oestradiol, insulin-like growth factor I). RESULTS: 80 subjects completed the trial. Daily milk intake at baseline averaged 150 ml in both groups. The intervention group consumed, on average, an additional 300 ml a day throughout the trial. Compared with the control group, the intervention group had greater increases of bone mineral density (9.6% v 8.5%, P = 0.017; repeated measures analysis of variance) and bone mineral content (27.0% v 24.1%, P = 0.009). No significant differences in increments in height, weight, lean body mass, and fat mass were observed between the groups. Bone turnover was not affected by milk supplementation. Serum concentrations of insulin-like growth factor I increased in the milk group compared with the control group (35% v 25%, P = 0.02). CONCLUSION: Increased milk consumption significantly enhances bone mineral acquisition in adolescent girls and could favourably modify attainment of peak bone mass. PMID- 9390051 TI - Birth defects in infants conceived by intracytoplasmic sperm injection: an alternative interpretation. AB - OBJECTIVE: To test the hypothesis that liveborn infants conceived by intracytoplasmic sperm injection are at an increased risk of having a major birth defect. DESIGN: Reclassification of the birth defects reported in infants born after intracytoplasmic sperm injection in Belgium and comparison with prevalence estimated in Western Australian population by means of same classification system. SETTING AND SUBJECTS: 420 liveborn infants who were conceived after intracytoplasmic sperm injection in Belgium and 100,454 liveborn infants in Western Australia delivered during the same period. MAIN OUTCOME MEASURES: Estimates of birth prevalence of birth defects and comparisons of odds ratios between cohort conceived after intracytoplasmic sperm injection and Western Australian infants. RESULTS: Infants born after intracytoplasmic sperm injection were twice as likely as Western Australian infants to have a major birth defect (odds ratio 2.03 (95% confidence interval 1.40 to 2.93); P = 0.0002) and nearly 50% more likely to have a minor defect (1.49 (0.48 to 4.66); P = 0.49). Secondary data-led analyses, to be interpreted with caution, found an excess of major cardiovascular defects (odds ratio 3.99), genitourinary defects (1.33), and gastrointestinal defects (1.84), in particular cleft palate (5.11) and diaphragmatic hernia (7.73). CONCLUSIONS: These results do not confirm the apparently reassuring results published by the Belgian researchers of intracytoplasmic sperm injection. Further research is clearly required. Meanwhile, doctors practising intracytoplasmic sperm injection should bear this alternative interpretation in mind when they counsel couples and obtain informed consent for the procedure. PMID- 9390053 TI - Variation in management of small invasive breast cancers detected on screening in the former south east Thames region: observational study. AB - OBJECTIVE: To examine the variation in surgical and adjuvant treatment of breast cancer of known histology and detected on screening in a large cohort of patients treated by the surgeons of a health region. DESIGN: Part prospective, part retrospective observational study using the databases of a region's breast screening programme and of the cancer registry. SETTING: The former South East Thames region. SUBJECTS: 600 women aged 49-79 who presented during 1991-2 with invasive breast cancer up to 20 mm in diameter that had been detected on screening. These patients were treated by 35 surgeons. MAIN OUTCOME MEASURES: Mastectomy rate by surgeon and the use of adjuvant treatment (radiotherapy, tamoxifen, and chemotherapy) were compared with risk factors, tumour grade, resection margins, and axillary node status. RESULTS: The mastectomy rate varied between nil and 80%, although the numbers at these extremes were small (0/13 v 8/10). Surgeons operating on more than 20 such cases had a lower mastectomy rate (15%) than surgeons treating fewer cases (23%), but this difference was confounded by variation in casemix. There were also wide variations in mastectomy rates and in axillary sampling rates that were independent of casemix or caseload. There was broad agreement on the use of adjuvant tamoxifen (94%), but few patients received chemotherapy (2.5%). 78 patients (19%) did not receive radiotherapy, including 51 out of 317 patients with unfavourable tumours, and 26 patients did not receive tamoxifen. Whether the patient received adjuvant treatment was more dependent on referral by the surgeon than the risk factors for local recurrence and was independent of caseload. CONCLUSION: Mastectomy rates for similar tumours vary widely by surgeon independently of casemix or caseload, but surgeons with a higher caseload tend to have a lower mastectomy rate. Omission of postoperative radiotherapy or tamoxifen after conservative treatment is not related to risk factors for local recurrence or caseload. Confidential feedback of treatment profiles to individual surgeons has been used, but when benefit has been established treatment should be guided by evidence based protocol. PMID- 9390054 TI - Is the SF-36 a valid measure of change in population health? Results from the Whitehall II Study. AB - OBJECTIVE: To measure within-person change in scores on the short form general health survey (SF-36) by age, sex, employment grade, and disease status. DESIGN: Longitudinal study with a mean of 36 months (range 23-59 months) follow up, with screening examination and questionnaire to detect physical and psychiatric morbidity. SETTING: 20 civil service departments originally located in London. PARTICIPANTS: 5070 male and 2197 female office based civil servants aged 39-63 years. MAIN OUTCOME MEASURES: Change in the eight scales of the SF-36 (adjusted for baseline score and length of follow up) and effect sizes (adjusted change standard deviation of differences). RESULTS: Within-person declines (worsening health) with age were greater than estimated by cross sectional data alone. General mental health showed greater declines among younger participants (P for linear trend < 0.001). Employment grade was inversely related to change; lower grades had greater deteriorations than higher grades (P < 0.001 for each scale in men; P < 0.05 for each scale in women except general health perceptions and role limitations due to physical problems). The greatest declines were seen among participants with disease at baseline, with the effects of physical and psychiatric morbidity being additive. Effect sizes ranged from 0.20 to 0.65 in participants with both physical and psychiatric morbidity. CONCLUSIONS: Health functioning, as measured by the SF-36, changed in hypothesised directions with age, employment grade, and disease status. These changes occurred within a short follow up period, in an occupational, high functioning cohort which has not been the subject of intervention, suggesting that the SF-36 is sensitive to changes in health in general populations. PMID- 9390055 TI - Deficient colour vision and interpretation of histopathology slides: cross sectional study. AB - OBJECTIVE: To determine whether histopathologists with deficient colour vision make more errors in slide interpretation than those with normal colour vision. DESIGN: Examination of projected transparencies of histopathological slides under standardised conditions by subjects whose colour discriminating ability was accurately assessed. SETTING: Departments of histopathology in 45 hospitals in the United Kingdom. SUBJECTS: 270 male histopathologists and medical laboratory scientific officers. MAIN OUTCOME MEASURES: Number of slides correctly identified by subjects whose colour vision was measured on the Ishihara, City University, and Farnsworth-Munsell 100 hue tests. RESULTS: Mean (SD) scores (out of 10) for doctors with colour deficient vision were 9.4 (0.7) v 9.9 (0.4) for controls (P < 0.01) and 7.5 (1.6) v 9.4 (0.7) for scientific officers (P < 0.001). When subjects with colour deficient vision were categorised into severe, moderate, or mild, there was a significant trend towards those with severe deficiency making more mistakes (P < 0.001). CONCLUSIONS: Histopathologists and medical laboratory scientific officers should have their colour vision tested; if they are found to have a severe protan or deutan deficiency, they should be advised to adopt a safe system of working. PMID- 9390056 TI - Prevalence of HIV-1 infection among heterosexual men and women attending genitourinary clinics in Scotland: unlinked anonymous testing. PMID- 9390057 TI - Omeprazole, H2 blockers, and polyarthralgia: case-control study. PMID- 9390058 TI - Challenges in managing dyspepsia in general practice. PMID- 9390059 TI - Science, medicine, and the future. Gene therapy. PMID- 9390060 TI - ABC of palliative care. Constipation and diarrhoea. PMID- 9390061 TI - Why are doctors ambivalent about patients who misuse alcohol? PMID- 9390063 TI - More widespread public debate on rationing is essential. PMID- 9390062 TI - Women's health. The childbearing years and after. AB - Health and development planners have tended to see women primarily in context of their reproductive role. As a result, solutions to women's health needs have been restricted to expanding and improving maternal and child health systems. There has recently been a major shift in direction, largely because of the influence of the world conference on population and development held in Cairo in 1994. Dr Guiseppe Benagiano, director of the special programme of research, development and research training in human reproduction based at the WHO, says, "We need to remind ourselves constantly that reproductive health is not simply a biomedical issue but one with serious implications for our general health and by extension, for all our efforts in human social and economic development." The 1993 world development report on health identified the lack of a clear strategy for engaging women in health care and suggested that child health services, prenatal care, treatment of sexually transmitted diseases, and family planning services should be provided jointly at convenient times. In an example of this, the Chilean Institute of Reproductive Medicine now offers integrated family planning services at the same time as child health services, and Thailand is experimenting with mobile health clinics to reach women in their homes. As the proportion of elderly women increases, old age is increasingly being seen as a female issue. With the impact of urbanisation and industrialisation, more of these women are living isolated lives, often suffering from chronic debilitating diseases. In his opening statement to the global commission on women's health in April 1995 which focused on health conditions of women in old age, Dr Hiroshi Nakajima, the WHO's director general, said: "Our goal should not be solely to extend lives in the physical sense, but to ensure that the added years are worth living." PMID- 9390064 TI - Patient's assessments of disability in multiple sclerosis. Most patients have difficulty in rating themselves on visual analogue scales. PMID- 9390065 TI - Screening people with a family history of cancer. Benefit of screening for ovarian cancer is unproved. PMID- 9390066 TI - Screening people with a family history of cancer. Taking a family history in primary care is important. PMID- 9390067 TI - Unsupervised surgical training. Logbooks are essential for assessing progress. PMID- 9390068 TI - Unsupervised surgical training. Surgical training teaches surgical method, surgical anatomy, and operative skills. PMID- 9390069 TI - Unsupervised surgical training. Other specialties can learn from level of supervision of surgical training. PMID- 9390070 TI - Discontinuation of cervical spine immobilisation. Immobilisation should not be discontinued in unconscious patients. PMID- 9390071 TI - Discontinuation of cervical spine immobilisation. In children, cord injury may be present despite normal radiographic appearances. PMID- 9390072 TI - Resuscitation. New advisory statements on life support have been published. PMID- 9390073 TI - Resuscitation. Resuscitation Council (UK) wants everyone who uses new recovery position to report experiences. PMID- 9390074 TI - Study to predict which elderly patients will fall shows difficulties in deriving and validating a model. PMID- 9390075 TI - Determining precise role of ethnicity in disease will be difficult. PMID- 9390077 TI - New connections between medical knowledge and patient care. Intervention of health professionals acts as an inductance, not as a resistor. PMID- 9390076 TI - New connections between medical knowledge and patient care. Human condition is full of decisions that aren't simple yes/no decisions. PMID- 9390078 TI - New connections between medical knowledge and patient care. Patients in most need of medical attention are least able to operate computers. PMID- 9390079 TI - New connections between medical knowledge and patient care. Information technology has much to offer certain aspects of health care. PMID- 9390080 TI - General practitioners want to know whether a treatment works and is safe in practice. PMID- 9390081 TI - Pigeon fancier's lung. Current methodology is not sensitive enough to monitor effectiveness of avoidance measures. PMID- 9390082 TI - Pigeon fancier's lung. Pigeon racers will not want to reduce time spent in lofts. PMID- 9390083 TI - Changing face of ectopic pregnancy. Medical treatment of ectopic pregnancy preserves reproductive potential. PMID- 9390084 TI - Changing face of ectopic pregnancy. Each centre should validate diagnostic algorithms for its own patients. PMID- 9390085 TI - Anaesthetists in Poland are on hunger strike. PMID- 9390086 TI - If a wound is "neatly incised" it is not a laceration. PMID- 9390087 TI - Multidrug resistant tuberculosis and HIV: a personal experience. PMID- 9390088 TI - Understanding deaf and hard-of-hearing patients. PMID- 9390089 TI - Different viewpoints on medical abortion. PMID- 9390090 TI - Different viewpoints on medical abortion. PMID- 9390091 TI - Different viewpoints on medical abortion. PMID- 9390092 TI - Different viewpoints on medical abortion. PMID- 9390093 TI - Venlafaxine. PMID- 9390094 TI - Practical guidelines for antepartum fetal surveillance. AB - Antepartum fetal assessment is used in pregnancies at high risk for perinatal morbidity and mortality. Current testing options include the fetal movement count, the nonstress test, the contraction stress test and the biophysical profile. Vibroacoustic stimulation is a useful adjunctive procedure. All of these modalities have limitations. A strict protocol for antepartum fetal surveillance that is applicable to all patients is not possible. However, a testing approach based on general principles and guidelines can be followed. PMID- 9390095 TI - Common cardiovascular problems in the young: Part II. Hypertension, hypercholesterolemia and preparticipation screening of athletes. AB - Blood pressure should be measured during health maintenance visits in all children three years of age and older. Cholesterol levels should be obtained in children with a family history of hypercholesterolemia or premature coronary artery disease and in children with other risk factors, such as hypertension, smoking or obesity. Preparticipation screening for sports participation should include a detailed questionnaire regarding the athlete's personal or family history of syncope, sudden death or arrhythmia, as well as measurement of blood pressure, auscultation of the heart and evaluation of upper and lower extremity pulses. PMID- 9390096 TI - Behavioral and pharmacologic treatment of delirium. AB - Delirium is an acute confusional state with a fluctuating course. Since the syndrome of delirium is associated with derangements of cognition, attention and level of consciousness, it can cause behaviors that are difficult to manage. Hallucinations, agitation, insomnia and anxiety are common in the delirious patient. Behavioral and pharmacologic interventions can be used while the underlying etiology of delirium is sought. Nonpharmacologic measures include frequent reassurance, environmental cues to reorient the patient and the judicious use of physical restraints. Haloperidol, which has negligible anticholinergic effects, is the drug most often used to treat the symptoms of delirium. Short-acting benzodiazepines may be useful in patients with delirium caused by alcohol withdrawal, but these agents may cause increased agitation in elderly patients and patients with hepatic dysfunction. PMID- 9390097 TI - Guidelines on the care of diabetic nephropathy, retinopathy and foot disease. AB - Diabetes mellitus is a common disease frequently managed by family physicians. Because of its high prevalence and associated comorbidity, diabetes mellitus has received a great deal of attention from several specialty organizations. The American Diabetes Association, the American Board of Family Practice and the Centers for Disease Control and Prevention have published specific practice guidelines and recommendations for the care of diabetic patients. These recommendations include annual comprehensive foot examinations, yearly ophthalmologic screening for retinopathy, and urinalysis for microalbuminuria. The use of angiotensin converting enzyme inhibitors is advocated for the majority of diabetic patients with proteinuria or hypertension. Based on recent evidence, improved glycemic control is also increasingly advocated. Compliance with practice guidelines by primary care physicians has historically been poor. Mechanisms such as the use of patient problem lists and diabetic flow sheets can serve as reminders to physicians and can facilitate closer adherence to practice guidelines. PMID- 9390098 TI - Drug treatment of migraine: Part I. Acute therapy and drug-rebound headache. AB - Most migraine patients need only abortive treatment for their headaches. By the time they present to a physician, they have already tried many over-the-counter medications for headache relief. Prioritizing treatment according to headache severity and associated symptoms will help the physician determine the most appropriate medications to use. Narcotics should be reserved for use only in patients unresponsive to adequate trials of non-narcotic agents. In some patients, the recurrent nausea and vomiting can be as disabling as the pain; antiemetic agents are an important adjunct to analgesic therapy in these patients. Sumatriptan and dihydroergotamine are more expensive than other migraine agents but have distinct therapeutic advantages in patients with moderate to severe headaches. Some patients experience rebound headache from overuse of analgesics and other headache medications. Educating patients about self-help measures and avoidance of triggers is an important element in the effective management of migraine headaches. PMID- 9390099 TI - Issues to consider in deaf and hard-of-hearing patients. The Committee on Disabilities of the Group for the Advancement of Psychiatry. AB - Successful interaction with patients who are deaf or hard of hearing requires an understanding of background issues, including the significance of the age of onset of deafness, the patient's choice of language, the patient's cultural identification and educational history, and the type of hearing loss. All of these factors should influence the physician's interview techniques and use of resources. PMID- 9390100 TI - Effect of maternal HIV-1 disease on infant outcome. PMID- 9390101 TI - ADA recommends a lower fasting glucose value in the diagnosis of diabetes mellitus. PMID- 9390102 TI - UCSF creates Web site for information on HIV/AIDS. PMID- 9390103 TI - Computational results may depend on personal computer processor. PMID- 9390104 TI - Apparent decrease in population clearance of theophylline: implications for dosage. AB - BACKGROUND: Having observed in recent years that the theophylline dose requirements needed to attain peak serum concentrations of 10 to 20 micrograms/ml infrequently reached previously described mean values, we hypothesized that a downward shift in the range of dose requirements had occurred among patients with asthma. STUDY DESIGN: We examined dosage requirements needed to attain peak serum concentrations of 10 to 20 micrograms/ml in all patients with chronic asthma treated with theophylline by the Pediatric Allergy and Pulmonary Clinic at the University of Iowa from 1990 to 1994 (n = 300) and at the Pediatric Pulmonary Clinic at the University of Florida from 1992 to 1995 (n = 93). We then compared these doses to previous dose requirements from 1978 to 1983 determined in the same manner. RESULTS: Despite similar mean peak serum concentrations during both time periods (14 micrograms/ml), mean theophylline dosage requirements during the period of this study were approximately 25% lower among all age groups than those previously observed (p < 0.001). There were no significant differences in mean dosage requirements between the Iowa and Florida patients in any age group examined. CONCLUSIONS: Theophylline dose requirements needed to attain serum concentrations of 10 to 20 micrograms/ml have decreased significantly from those on which current dosing recommendations are based. This suggests a decrease in mean clearance of the population. PMID- 9390105 TI - Influence of stiripentol on cytochrome P450-mediated metabolic pathways in humans: in vitro and in vivo comparison and calculation of in vivo inhibition constants. AB - OBJECTIVE: The spectrum of cytochrome P450 inhibition of stiripentol, a new anticonvulsant, was characterized in vitro and in vivo. METHODS: Stiripentol was incubated in vitro with (R)-warfarin, coumarin, (S)-warfarin, (S)-mephenytoin, bufuralol, p-nitrophenol, and carbamazepine as probes for CYPs 1A2, 2A6, 2C9, 2C19, 2D6, 2E1, and 3A4, respectively. Caffeine demethylation and the 6 beta hydroxycortisol/cortisol ratio were monitored in vivo before and after 14 days of treatment with stiripentol as measures of CYP1A2 and CYP3A4 activity, and dextromethorphan O- and N-demethylation were used to measure CYP2D6 and CYP3A4 activity, respectively. In vivo inhibition constants for CYP3A4 were calculated with use of data that previously documented the interaction between stripentol and carbamazepine. RESULTS: In vitro, stiripentol inhibited CYPs 1A2, 2C9, 2C19, 2D6, and 3A4, with inhibition constant values at or slightly higher than therapeutic (total) concentrations of stiripentol, but it did not inhibit CYPs 2A6 and 2E1 even at tenfold therapeutic concentrations. In vivo inhibition of caffeine demethylation and dextromethorphan N-demethylation were consistent with inhibition of CYP1A2 and CYP3A4, respectively. The 6 beta hydroxycortisol/cortisol ratio did not provide a reliable index of CYP3A4 inhibition. Inhibition of CYP2D6-mediated O-demethylation was not observed in vivo. With use of carbamazepine, in vivo inhibition constants for CYP3A4 ranged between 12 and 35 mumol/L, whereas the corresponding in vitro value was 80 mumol/L. CONCLUSIONS: Stiripentol appears to inhibit several CYP450 enzymes in vitro and in vivo. In vivo inhibition constants show that stiripentol inhibition of CYP3A4 is linearly related to plasma concentration in patients with epilepsy. PMID- 9390106 TI - Impact of long-term ethanol consumption on CYP1A2 activity. AB - Ethanol is a well-known inducer of CYP2E1; whether or not it is an inducer of other cytochromes has not been investigated systematically. The aim of our study was to evaluate the impact of ethanol consumption on the activity of CYP1A2, which has been shown to be influenced by drugs (inhibited or induced). We evaluated CYP1A2 activity by the ratio of the molar urinary concentrations of the three end products of paraxanthine demethylation of caffeine to the molar concentration of a paraxanthine 8-hydroxylation product. This urinary metabolite ratio has previously been shown to correlate with caffeine clearance. The caffeine metabolites were measured in urine collected during the 3 hours after oral administration of 200 mg caffeine. The caffeine test was performed in 12 smokers (> 25 cigarettes/day) and 12 nonsmokers, all of whom were alcoholic inpatients (daily intake > 100 mg absolute ethanol), within the first 3 days of their hospital stay and after 14 days of abstinence from ethanol. In alcoholic patients who were smokers the molar urinary concentration ratio was 3.14 +/- 0.97 before withdrawal and 4.01 +/- 0.92 after 14 days of abstinence from ethanol. In contrast, in alcoholic patients who were nonsmokers it was 2.62 +/- 0.95 and 2.18 +/- 0.96 before and after withdrawal, respectively. In volunteers who were smokers the molar urinary concentration ratio was 5.02 +/- 1.51, whereas in volunteers who were nonsmokers it was 3.22 +/- 1.46. Our results confirm the well known induction of CYP1A2 activity by tobacco smoking and show that this induction is masked by long-term ethanol consumption. PMID- 9390107 TI - Itraconazole increases plasma concentrations of quinidine. AB - BACKGROUND: Quinidine is eliminated mainly by CYP3A4-mediated metabolism. Itraconazole interacts with some but not all of the substrates of CYP3A4; it is therefore important to study the possible interaction of itraconazole with quinidine. METHODS: A double-blind, randomized, two-phase crossover study design was used with nine healthy volunteers. Itraconazole (200 mg) or placebo was ingested once a day for 4 days. A single 100 mg oral dose of quinidine sulfate was ingested on day 4. Plasma concentrations of quinidine, itraconazole, and hydroxyitraconazole, as well as cumulative excretion of quinidine into urine, were determined up to 24 hours. The ECG, heart rate, and blood pressure were also recorded up to 24 hours. RESULTS: On average the peak plasma concentration of quinidine increased to 1.6-fold (p < 0.05), and the area under the concentration time curve of quinidine increased to 2.4-fold (p < 0.01) by itraconazole. The elimination half-life of quinidine was prolonged 1.6-fold (p < 0.001), and the area under the 3-hydroxyquinidine/quinidine ratio-time curve decreased to one fifth (p < 0.001) by itraconazole. The renal clearance of quinidine decreased 50% (p < 0.001) by itraconazole, whereas the creatinine clearance was unaffected. The QTc interval correlated with the concentrations of quinidine during both itraconazole and placebo phases (r2 = 0.71 and r2 = 0.79, respectively; p < 0.01), although only minor changes between the phases were observed in other pharmacodynamic variables. CONCLUSIONS: Itraconazole increases plasma concentrations of oral quinidine, probably by inhibiting the CYP3A4 isozyme during the first-pass and elimination phases of quinidine. The decreased renal clearance of quinidine might be the result of the inhibition of P-glycoprotein mediated tubular secretion of quinidine by itraconazole. The concentrations of quinidine should be closely monitored if itraconazole or some other potent CYP3A inhibitors are used with quinidine. PMID- 9390108 TI - Population pharmacokinetics of riluzole in patients with amyotrophic lateral sclerosis. AB - OBJECTIVES: To characterize the population pharmacokinetic of riluzole in patients with amyotrophic lateral sclerosis (ALS). METHODS: One hundred patients with ALS who were participating in a multicenter phase III dose-ranging trial of riluzole were sampled on 179 visits. The sampling strategy (two samples per visit) was varied across patients to define the population kinetic profile (full screen). Riluzole plasma levels were determined by HPLC, and the data were analyzed by nonlinear mixed-effect modeling (NONMEM program) with use of a one compartment structural model. The model incorporated interoccasion (visit-to visit) variability. RESULTS: In the basic one-compartment pharmacokinetic model, interindividual variability in plasma clearance (51.4%) was higher than intraindividual (visit-to-visit) variability (28.0%), indicating uniform pharmacokinetic behavior during long-term therapy. Riluzole clearance was independent of dosage (25 to 100 mg twice daily), treatment duration (up to 10 months), age, and renal function; gender and smoking were the most important patient covariates, with hepatic function having lesser influence. Typical value of clearance was 51.4 L/hr for a nonsmoking male patient. It was 32% lower in women than in men and 36% lower in nonsmokers than in smokers. Gender- and smoking-related variations in riluzole exposure at the recommended dosage (50 mg twice daily) were within the range of exposures achieved (with no untoward effect) in this dose-ranging study. CONCLUSION: The pharmacokinetics of riluzole has been characterized in patients during long-term therapy. Riluzole clearance is independent of dose and treatment duration. Within-patient variability is low. Gender and smoking status are the main covariates to explain interpatient variability. PMID- 9390109 TI - Mephenytoin disposition and serum bile acids as indices of hepatic function in chronic viral hepatitis. AB - BACKGROUND AND OBJECTIVES: The effect of chronic viral hepatitis on liver function may vary from none to hepatic failure. Changes in function are usually the result of impaired hepatocyte function or altered vascular flow and architecture. Conventional liver function tests usually cannot distinguish contributions from these mechanisms or indicate degree of hepatic metabolic dysfunction. An alternative approach is to measure the hepatic metabolism of a highly extracted compound whose oral clearance and systemic bioavailability are dependent on both hepatocyte function and degree of portosystemic shunt. METHODS: The stereoselective metabolism of racemic mephenytoin (100 mg oral dose) was investigated in 35 patients with chronic viral hepatitis and compared with 153 healthy subjects. The mephenytoin R/S enantiomeric ratio and cumulative excretion of the 4'-hydroxymephenytoin metabolite in a 0- to 8-hour urine sample were used in addition to serum bile acid levels and pathologic examination of biopsy specimens to assess the severity of hepatic dysfunction and portosystemic shunting. RESULTS: The patients as a group excreted less 4'-hydroxymephenytoin and had a smaller R/S enantiomeric ratio of mephenytoin. The two measures were discriminatory between the patient groups classified by either serum cholylglycine level or pathologic examination of biopsy specimens. Combination of the two measures of mephenytoin metabolism allowed the patients to be classified into three groups: normal hepatocyte function without portosystemic shunt, normal hepatocyte function with portosystemic shunt, and low hepatocyte function with or without portosystemic shunt. CONCLUSION: This study has shown the potential usefulness of mephenytoin metabolism as a sensitive indicator of hepatic pathologic condition with an ability to discriminate between contributory alternative mechanisms. PMID- 9390110 TI - Lack of cross-tolerance to short-term linsidomine in forearm resistance vessels and dorsal hand veins in subjects with nitroglycerin tolerance. AB - BACKGROUND: Therapy with nitroglycerin is widely used in the treatment of angina pectoris, but development of tolerance is a major problem. Nitrovasodilators other than nitroglycerin may be less prone to induce vascular tolerance. This investigation was designed to test whether the alternative nitric oxide donor linsidomine maintains its vasodilator effects in the presence of nitroglycerin tolerance. METHODS: We tested the vascular effects of nitroglycerin and linsidomine (SIN-1) in forearm resistance arteries (venous occlusion plethysmography) and hand veins (venous compliance technique) using a randomized, double-blind placebo-controlled regimen in 33 healthy subjects (age range, 22 to 38 years; mean age, 26 years) before and after 7 days of assignment to either 1 week of nitroglycerin administration (0.83 mg/hr) for induction of tolerance or placebo administration. RESULTS: Vascular responses of both vascular beds to nitroglycerin (in veins: mean difference, 42.3%; confidence interval [CI], 3% to 81.7%; p < 0.05; in arteries: mean difference, 65.0%; CI, 38.9% to 91.1%; p < 0.01) but not to linsidomine (in veins: mean difference, -13.8%; CI, -53.5 to 25.8%; not significant; in arteries: -19.7%; CI, -33.7% to -5.6%; not significant) were attenuated in the nitroglycerin patch group, whereas the placebo group showed no differences to either nitroglycerin (in arteries: mean difference, -7.5%; CI, -44.6% to 29.6%; in veins: -10.6%; CI, -58.2% to 36.9%) or linsidomine (in arteries: 4.5%; CI, -12.8% to 21.7%; in veins: -13.1%; CI, -4.5% to 29.8%). CONCLUSION: These results suggest that short-term administration of sydnonimines can overcome the loss of vascular relaxation associated with long term nitroglycerin therapy. PMID- 9390111 TI - Clinical pharmacodynamics of SDZ HTF 919, a new 5-HT4 receptor agonist, in a model of slow colonic transit. AB - OBJECTIVES: To explore the pharmacodynamic effects of the new promotile agent SDZ HTF 919, a selective partial 5-HT4 receptor agonist, in healthy subjects. METHODS: A pharmacodynamic model was applied to prolong colonic transit by dietary means. Subsequently, the effects of twice-daily multiple doses of SDZ HTF 919 (1, 5, 25, and 100 mg) were investigated in a randomized, double-blind, placebo-controlled parallel-group study with 12 subjects per dose level. The sequential design with three study periods of 7 days each included intake of a self-selected diet, a liquid formula diet with soluble fiber supplementation, and a fiber-supplemented diet together with either SDZ HTF 919 or placebo administration. Stool characteristics (frequency and consistency) and total colonic transit times (with use of radiopaque markers) were recorded in each study period. RESULTS: SDZ HTF 919 was well tolerated at all dose levels. The frequency of loose stool and headache increased with higher doses. After a fiber supplemented diet intake, the median stool frequency decreased from 8 1/2-9 to 5 7 defecations per study period. SDZ HTF 919 in doses of 25 and 100 mg twice a day increased the stool frequency (p < 0.05). Stool consistency was softened by all but the lowest SDZ HTF 919 dose. A fiber-supplemented diet prolonged total colonic transit time in all groups by 45 hours on average. Twice-a-day administration of SDZ HTF 919 for 6 days in addition to a fiber-supplemented diet significantly shortened the total colonic transit time only at the 5 mg dose. The lack of effect at lower and higher SDZ HTF 919 doses suggests a biphasic dose response relationship for total colonic transit time. CONCLUSIONS: The suitability of total colonic transit time measurements in healthy subjects as a surrogate marker should be confirmed by patient studies. PMID- 9390112 TI - Dose-related protection of exercise bronchoconstriction by montelukast, a cysteinyl leukotriene-receptor antagonist, at the end of a once-daily dosing interval. AB - The dose-related protective effects of montelukast, a potent and selective cysteinyl leukotriene-receptor antagonist, against exercise-induced bronchoconstriction were investigated in a five-period, randomized, incomplete block, crossover study with montelukast (0.4, 2, 10, 50 mg) and placebo. The study subjects were 27 nonsmoking, healthy stable patients with asthma (mean forced expiratory volume in 1 second [FEV1], 82.0% predicted) who demonstrated a > or = 20% decrease in FEV1 while beta-agonist was withheld for 6 hours before treadmill exercise. The standard exercise challenge was performed 20 to 24 hours, and again 32 to 36 hours, after the second of two once-daily doses. The effect of oral montelukast on exercise was measured by the area above the postexercise percentage decrease in FEV1 versus time curve from 0 to 60 minutes [AUC(0-60)], the maximal percentage decrease in FEV1 after exercise, and time after maximal decrease to recovery of FEV1 to within 5% of the preexercise baseline. Twenty to 24 hours after administration, montelukast caused dose-related protection, while providing similar protection against exercise-induced bronchoconstriction at the two highest doses. The AUC(0-60) values (mean +/- SD) were 637 +/- 898, 715 +/- 870, 988 +/- 1147, and 927 +/- 968 min. % for 50, 10, 2, and 0.4 mg montelukast, respectively, and 1193 +/- 1097 min. % for placebo (p = 0.003). No important clinical effect was present 36 hours after dosing. Montelukast was generally well tolerated at all dose levels. In conclusion, montelukast caused dose-related protection against exercise-induced bronchoconstriction at the end of a once daily dosing interval. Protection against exercise-induced bronchoconstriction can be used to determine appropriate dose selection. PMID- 9390113 TI - Dexamethasone down-regulates the expression of L-selectin on the surface of neutrophils and lymphocytes in humans. AB - OBJECTIVE: On the basis of previous animal studies, we hypothesized that dexamethasone may reduce the expression of L-selectin on neutrophils and lymphocytes in healthy men. METHODS: A double-blind, randomized, placebo controlled, and three-way crossover trial was conducted in nine healthy men. Every subject received four identical infusions of saline solution, 0.04 mg/kg dexamethasone, or 1.0 mg/kg dexamethasone during three observation periods of 48 hours each. RESULTS: Dexamethasone time and dose dependently decreased the L selectin expression on neutrophils and lymphocytes as measured by flowcytometry. This effect occurred with a time lag of 8 hours after start of treatment: the L selectin binding index of neutrophils decreased by a maximum of -50% (confidence interval [CI], -37% to -63%) and that of lymphocytes by -26% (CI, -8% to -45%) at 32 hours after the start of treatment with high-dose dexamethasone (p < 0.016). Low-dose dexamethasone had only a transient effect on L-selectin expression of lymphocytes and a less pronounced effect on L-selectin expression of neutrophils. CONCLUSION: Dexamethasone time and dose dependently decreases L-selectin expression on neutrophils and lymphocytes in health men, an effect that is less pronounced than that previously reported for animals. PMID- 9390114 TI - Buprenorphine withdrawal syndrome in a newborn. AB - A pregnant woman who was addicted to heroin rapidly withdrew from illicit drugs after the onset of a 4 mg/day buprenorphine treatment. In the newborn's blood, urine, and meconium 20 hours after birth, high concentrations of buprenorphine and its metabolite norbuprenorphine were detected, with a higher buprenorphine/norbuprenorphine ratio than in adults, possibly as a consequence of immature hepatic function; no illicit drugs were found. The child had a weak withdrawal syndrome on the second day of life and recovered rapidly. The measured buprenorphine daily dose ingested by the newborn through mother's milk was very low (3.28 micrograms) and probably had little pharmacologic effect because no withdrawal signs could be noted when maternal feeding was later abruptly interrupted. Further investigations are required to determine whether buprenorphine can be considered to be a good alternative to methadone in the treatment of pregnant heroin addicts to prevent marked withdrawal syndromes in newborns. PMID- 9390115 TI - Ticlopidine inhibition of phenytoin metabolism mediated by potent inhibition of CYP2C19. AB - A patient who had taken a stable dose of phenytoin for 2 years had a coronary stent placed for unstable angina and ticlopidine was added to his therapeutic regimen. Twenty-five days later, he was hospitalized with acute symptomatic phenytoin toxicity and a serum concentration of 46.5 micrograms/ml. Determination of metabolic genotype revealed that the patient had a wild-type genotype for CYP2C9, CYP2C19, and CYP2D6. Using human liver microsomes, we showed that ticlopidine is a potent inhibitor of cytochrome P450 2C19, with an estimated inhibition constant (Ki) of 3.7 +/- 0.2 mumol/L. The influence of ticlopidine on CYP2C9, the other cytochrome P450 isoform that metabolizes phenytoin, is relatively weak, with a calculated Ki of 38.8 +/- 27 mumol/L. These data suggest that, in this patient, phenytoin toxicity was caused by inhibition of CYP2C19 by ticlopidine, and the data emphasize the importance of CYP2C19 in the metabolism of phenytoin. PMID- 9390116 TI - Informing the public. PMID- 9390117 TI - Toxicity of lidocaine desethyl metabolites. PMID- 9390118 TI - Endoscopic ultrasound-guided fine needle aspiration. PMID- 9390119 TI - Clinical utility of endoscopic ultrasound-guided fine needle aspiration. AB - OBJECTIVE: To assess our institution's initial experience with the clinical utility of endoscopic ultrasound (EUS)-guided fine needle aspiration. STUDY DESIGN: Prospective analysis of the clinical utility of EUS-guided FNA. RESULTS: Fifty-three patients underwent EUS-guided FNA of 64 sites, 28 for pancreatic masses, 15 for lymph nodes, 10 for solid lesions, 7 for cystic masses, 2 for submucosal masses and 2 for perigastrointestinal fluid. A cytopathologist was present during all procedures. An average of four passes (range, one to nine) was required to make a diagnosis in the 22 patients with pancreatic malignancies. There was one possible complication among the 53 patients. In 36 of the 53 patients, the combination of diagnostic EUS findings and cytologic diagnosis made a major change in the patient's management. CONCLUSION: Because of its ability to affect patient management, EUS-guided FNA will become a more commonly used procedure, especially at oncologic centers. Since the number of fine needle passes needed for diagnosis is quite variable, it is important to have a cytopathologist participate in these procedures. PMID- 9390120 TI - Ultrasound-guided fine needle aspiration cytology of malignant gallbladder masses. AB - OBJECTIVE: To determine the accuracy and reliability of ultrasound (US)-guided fine needle aspiration cytology (FNAC) over blind aspiration in gallbladder masses. STUDY DESIGN: We performed FNAC in 107 cases of carcinoma of the gallbladder; blind aspiration was done in 71 patients (66.36%) and US-guided aspiration in 36 (33.64%). In cases where FNAC after the first aspiration showed the aspirate to be inflammatory, acellular (inconclusive) or suspicious for malignancy, FNAC was repeated under US guidance. Diagnosis was later confirmed by histopathology in all cases. RESULTS: After the first aspiration, gallbladder malignancy was confirmed in 77 (71.96%) cases. Of these 77 cases, 34 underwent US guided aspiration, and the remaining 43 underwent blind aspiration. Cases with inflammatory or acellular (inconclusive) aspirates or that were suspicious for malignancy after the first aspiration underwent a second aspiration under ultrasonic guidance. On the second aspiration of 30 cases, 16 (53.33%) proved to be of adenocarcinoma, 7 (23.33%) were suspicious for malignancy, 5 (16.66%) were inflammatory, and 2 (6.66%) were acellular. Diagnosis was later confirmed by histopathology in all cases. US-guided FNAC had diagnostic accuracy of 95% as compared to 60% on blind aspiration. There was no major complication or needle tract recurrence of the disease. CONCLUSION: US-guided FNAC is safe, rapid, reliable, cost-effective and accurate in diagnosing gallbladder carcinoma. PMID- 9390121 TI - Identifying cytologic characteristics and grading endocervical columnar cell abnormalities. A study aided by high-definition television. AB - OBJECTIVE: To test the ability of cytotechnologists to recognize and accurately interpret selected architectural, cellular and nuclear features presented on a high-definition television (HDTV) and to make a reliable diagnosis with HDTV. STUDY DESIGN: A total of 1,122 features considered diagnostic of different endocervical columnar cell abnormalities were selected from 50 smears from 48 women with the help of a motor-driven-stage microscope by five observers who had knowledge of the final diagnosis. The selected and stored features were presented on an HDTV and evaluated in five successive sessions without knowledge of the final diagnosis. RESULTS: Specific types of features were correctly identified in a high number of cases. Considerable interobserver variability was demonstrated in the scoring of grades of expression of features. Overrated and under-rated monitor diagnoses were related to overvalued and undervalued features. From a group of 437 images that were correctly diagnosed by four or five observers, five features proved to be highly related to the correct diagnosis. CONCLUSION: Observers were capable of making a reliable diagnosis on features, selected by other observers, when presented on an HDTV. An overall correct diagnosis was made in 93% of cases. PMID- 9390122 TI - Atypical squamous cells of undetermined significance. A cytohistologic study of 52 cases. AB - OBJECTIVE: To evaluate the significance of atypical squamous cells of undetermined significance (ASCUS) by correlating the histologic findings following a diagnosis of ASCUS on a cervical cytologic smear. STUDY DESIGN: Eighty-four smears that had been called ASCUS over a five-month period and that had corresponding histologic material were reviewed independently. Only 52 of the 84 cases on which a consensus was reached were retained for the current study. RESULTS: The breakdown of the follow-up histologic diagnoses was as follows: 28 cases (54%) were negative (without squamous intraepithelial lesions [SIL]); 22 cases (42%) showed SILs, of which 14 (27%) were low grade, 5 (10%) were high grade and 3 (5%) had SILs that could not be further classified because of fragmentation of the endocervical curettings. Finally, two cases (4%) proved to be invasive cervical carcinoma on histology despite smears that were satisfactory and not limited by the quantity or quality of material; in these the discrepancy was attributed to sampling error. CONCLUSION: Patients whose cervical cytologic smears fall into the category of ASCUS may, on follow-up, exhibit a wide spectrum of findings, ranging from no pathologic abnormality to frequent SIL and even to invasive carcinoma in rare instances. A diagnosis of ASCUS on smears warrants careful follow-up and investigation. PMID- 9390123 TI - Cervical smear histories of 585 women with biopsy-proven carcinoma in situ. AB - OBJECTIVE: To analyze the cervical cytologic smear history of women with carcinoma in situ (CIS). STUDY DESIGN: We examined cytologic smears obtained within the three-year period prior to a histologic diagnosis of CIS in 585 women for whom at least one prebiopsy smear was available. RESULTS: Among 454 patients with only one smear available for review, 9 (2%) had a negative cytologic diagnosis, 58 (13%) had low grade squamous intraepithelial lesion (LSIL), and 387 (85%) had high grade squamous intraepithelial lesion (HSIL). One hundred thirty one women had two to five smears taken within the previous three years available for review. All the smears taken prior to biopsy showed HSIL. The original diagnosis on the other smears was negative for 78 women (60%), HSIL for 46 (35%) and LSIL for 7 (5%). ALl 132 smears originally classified as negative from 87 of 585 (14.8%) women were reviewed. Twenty-seven (20%) were then classified as showing HSIL, 10 as LSIL, 10 as atypical squamous cells of undetermined significance, 7 as unsatisfactory and 78 (59%) as remaining negative. CONCLUSION: Of smears classified as negative and taken in the three years before biopsy proven CIS, 41% were reclassified, with half reclassified as showing HSIL. PMID- 9390124 TI - Wisconsin Cytology Proficiency Testing Program. Results of voluntary testing in 1994. AB - OBJECTIVE: The Wisconsin Cytology Proficiency Testing Program (WCPTP) was developed cooperatively by the Wisconsin State Laboratory of Hygiene, the Wisconsin Society of Pathologists and the Wisconsin Society of Cytology to enable pathologists and cytotechnologists in Wisconsin to meet Clinical Laboratory Improvement Act of 1988 (CLIA '88) requirements for proficiency testing (PT). STUDY DESIGN: A joint steering committee designed the WCPTP to comply with all CLIA '88 regulations. The WCPTP application to the Health Care Financing Administration received tentative approval in May 1994. In 1994, mock PT was conducted at meetings of both state societies, and voluntary, on-site PT was conducted at 19 laboratories. RESULTS: Each of the 119 participants (49 pathologists, 70 cytotechnologists) was tested with sets of 10 glass slides, each representing one of four specified categories: A, unsatisfactory; B, normal/benign; C, low grade squamous intraepithelial lesion; and D, high grade squamous intraepithelial lesion and cancer. The failure rate for pathologists was 22.5% (11/49) and for cytotechnologists, 1.4% (1/70). The CLIA '88 scoring system for pathologists is more stringent. If cytotechnologists were scored as pathologists, 10% (7/70) would have failed. Using the cytotechnologist grid, 14.5% (7/49) of the pathologists would have failed. CONCLUSION: This voluntary program provided some preliminary insights into the issues related to PT evaluation of personnel competence and diagnostic criteria. PMID- 9390125 TI - Cervical cytologic smear false negative fraction. Reduction in a small community hospital. AB - OBJECTIVE: To determine the false negative fraction (FNF) at a small community hospital and its relation to the discovery of a significant error. STUDY DESIGN: All cervical cytologic smears (6,889) initially interpreted over a one-year period (1992) as "normal" or "near normal" were retrospectively rescreened and interpreted by outside institutions, without knowledge of the initial interpretation, to calculate yearly and quarterly FNFs. RESULTS: The overall FNF for 1992 was 12.3% and was 19.1%, 22.2%, 3.8% and 6.1% per successive quarters in 1992. A significant error was discovered at the start of the third quarter that subsequently received both local and national media attention. CONCLUSION: This study gives further proof that the FNF can be reduced to < 5% by motivated cytotechnologist/ pathologist teams, although it may not be possible to maintain this low an FNF. PMID- 9390126 TI - False positive cervicovaginal cytology. A follow-up study. AB - OBJECTIVE: To determine what percentage of cervical cytologic diagnoses initially classified as false positives (based on a negative cervical biopsy within three months of the cervical cytologic smear) are recategorized as histologic false negatives when subsequent studies reveal abnormalities. STUDY DESIGN: A three year review of 1,242 cervicovaginal biopsies with corresponding cytology in the preceding three months revealed 68 cases (5.5%) where the cytology was positive for a squamous intraepithelial lesion but the biopsy was within normal limits or showed benign cellular changes. Follow-up cytologic and/or histologic diagnoses were obtained for 53 of the 68 cases from the patients' hospital and physician office records. RESULTS: Of the 53 cases with follow-up, 24 (45%) were found to have a subsequent squamous intraepithelial lesion (indicating a sampling error at the time of the initial biopsy), and 9 showed atypical squamous cells of undetermined significance. In addition, 9 of the 20 patients (45%) who had negative follow-up studies had benign abnormalities on the initial, noncorrelating biopsy that may have contributed to the discrepancy. CONCLUSION: This study emphasized the importance of diligent follow-up of patients with noncorrelating studies since they represent a population at high risk for the subsequent detection of premalignant conditions. PMID- 9390127 TI - Glandular cells in vaginal smears from posthysterectomy patients. AB - OBJECTIVE: To assess the significance of nonatypical glandular cells in vaginal smears from patients who had undergone total hysterectomy. STUDY DESIGN: Vaginal smears with nonatypical glandular epithelium obtained from post-total hysterectomy patients were identified in our files over a 4.5-year period. The cytologic findings were correlated with the clinical data. RESULTS: Smears with nonatypical glandular epithelium from 15 post-total hysterectomy patients were identified, making this the largest series in the literature. The patients' mean age was 59 years. Most patients (73%) had a history of gynecologic malignancy, and 60% had received radiotherapy. All patients had a normal gynecologic examination when the vaginal smear was obtained. None of the patients developed recurrent or de novo vaginal adenocarcinoma. CONCLUSION: The presence of nonatypical glandular epithelial cells in smears from total hysterectomy patients is not indicative of adenocarcinoma. PMID- 9390128 TI - Diagnostic role of testicular fine needle aspiration biopsy in male infertility. AB - OBJECTIVE: To study the diagnostic role of fine needle aspiration biopsy (FNAB) of the testis in male infertility. STUDY DESIGN: A retrospective study of 586 cases of infertile males with oligospermia and azoospermia. The material obtained was stained with Diff-Quik. The proportion of Sertoli cells versus spermatogenic cells was studied. RESULTS: Cytologic examination revealed normal spermatogenesis in 10.2%, hypospermatogenesis in 31.4%, Sertoli cells only in 30.2% and an atrophic pattern in 28.6%. CONCLUSION: The patterns recognized by FNAB were comparable to those obtained by open biopsy. However, FNAB is less invasive, with very few complications. The procedure was well tolerated by all patients. There were very few complications. The findings of this study support the contention that FNAB of the testis is a reliable, relatively noninvasive procedure that has an important role in male infertility. PMID- 9390129 TI - Cytomorphology of tyrosine-rich crystalloids in fine needle aspirates of salivary gland adenomas. AB - OBJECTIVE: To evaluate the presence of tyrosine-rich crystalloids (TRC) in fine needle aspiration (FNA) specimens of pleomorphic adenomas of salivary gland. STUDY DESIGN: FNA specimens from 12 patients were reviewed, and the percentage of cases showing TRC was established. The staining properties of the TRC were evaluated as well as spontaneous fluorescence under ultraviolet (UV) light. RESULTS: Of the 12 pleomorphic adenomas, 4 showed TRC (30%) in the smears. Among the eight cytologically negative cases there were two that showed a few TRCs on histology. All positive cases were from African American patients. TRC stained weakly with Papanicolaou stain. TRC were deep blue with Diff-Quik. They fluoresced under UV light. CONCLUSION: TRC could be detected in FNA specimens. They were best seen under UV light. The Papanicolaou technique stained TRC very pale, making them difficult to see. Diff-Quik stained TRC dark blue, mimicking deposits of dye. The amount of TRC in histology paralleled the detection rate in cytology. PMID- 9390130 TI - P53 protein expression and DNA ploidy in common epithelial tumors of the ovary. AB - OBJECTIVE: To investigate p53 protein expression and DNA content in imprints from surgical biopsies of common epithelial tumors of the ovary. STUDY DESIGN: The study was based on 60 cases of epithelial tumors of the ovary (15 benign, 3 border-line and 42 malignant). For the demonstration of p53 protein, immunocytochemical staining with the avidin-extravidin technique was performed using monoclonal antibody p53 DO-7. DNA content was measured by image cytometry after Feulgen staining. RESULTS: There was a strong correlation between p53 expression and aneuploidy, with the difference between diploid and aneuploid tumors statistically significant (P < .001). A correlation was found between DNA ploidy, histologic grade and clinical stage (P < .001 and P < .05), respectively. There was no correlation between DNA ploidy and histologic type (P = .89). No correlation was observed between p53 protein expression and grade or clinical stage of the tumors. Nevertheless, a correlation of p53 expression between early (I, II) and advanced stages (III, IV) (P < .05) was observed. All benign and borderline tumors were diploid and did not express p53 protein. CONCLUSION: The results of the present study and the data in the literature stress the value of p53 expression and DNA ploidy in assessing the malignant potential of common epithelial ovarian cancers. However, the clinical application of these data requires further study. PMID- 9390131 TI - Diagnostic value of p53 protein and flow cytometric DNA analysis in the study of serous effusions. AB - OBJECTIVE: To investigate the diagnostic value of p53 protein and DNA analysis in the study of serous effusions. STUDY DESIGN: A total of 76 samples of serous effusions were studied by immunohistochemistry for p53 protein and flow cytometric (FCM) DNA analysis. The results were correlated with final cytologic diagnoses, which were confirmed by immunohistochemistry using antibodies against cytokeratin, carcinoembryonic antigen, epithelial membrane antigen and fibronectin. RESULTS: Final cytologic diagnoses included 28 malignant effusions and 48 benign effusions. No expression of p53 protein was seen in benign effusions. In contrast, p53 protein expression was seen in 19/28 (sensitivity 68%) malignant effusions. FCM detected aneuploid cells in 12/28 (43% sensitivity) of malignant and 0/46 of benign effusions. Immunohistochemical determination of p53 protein combined with FCM DNA analysis increased sensitivity to 79%. CONCLUSION: Immunohistochemical determination of p53 protein and FCM DNA analysis can aid in making an accurate and specific diagnosis of serous effusions, but the principal limitation of these tests is their relatively low sensitivity. PMID- 9390132 TI - Nuclear grooves in ependymoma. Cytologic study of 21 cases. AB - OBJECTIVE: To study the efficacy of crush preparation smears in the diagnosis of ependymomas. STUDY DESIGN: The study group consisted of 21 patients aged 7-61 years. All were admitted to Shiraz University hospitals (Nemazi and Beheshti) with intramedullary tumors. Fourteen were ventricular (fourth ventricle), 1 was in the parietal lobe, 5 were in the lumbosacral region and 1 was in the cauda equina. Intraoperative crush preparation smears were obtained from tissue, which was sent for frozen section and diagnosed cytologically. The control group consisted of 123 intracranial tumors (meningiomas, schwannomas, astrocytomas, oligodendrogliomas, medulloblastomas, pituitary adenomas, choroid plexus papilloma, craniopharyngioma and metastatic tumors). RESULTS: The smears in 11 cases revealed perivascular pseudorosettes, and the smears in 21 cases revealed ependymal rosettes. Papillary clusters, calcification and intranuclear inclusions were seen in two cases. Acinar structures were seen in seven cases. Myxomatous material was seen in one case. Nuclear grooves were seen in 15 cases. All cases were diagnosed as ependymomas. Biopsy specimens confirmed the cytologic diagnosis. The tumors in the control group showed no evidence of nuclear grooves. CONCLUSION: Fifteen cases of ependymoma showed a substantial number of nuclear grooves. Intraoperative crush preparation smears were very useful in the diagnosis of ependymomas and helped with the rapid interpretation of frozen sections. PMID- 9390133 TI - Urine cytology and the diagnosis of renal allograft rejection. I. Studies using conventional staining. AB - OBJECTIVE: To determine the reproducibility and validity of urine cytology for the diagnosis of acute renal allograft rejection (AR). STUDY DESIGN: We conducted a blind, prospective study of 10 renal allograft recipients. Freshly voided aliquots of urine were obtained on each hospital day and at each outpatient visit for a mean of 52.8 +/- 26.2 (SD) days following transplantation. The samples were prepared by cytocentrifugation and then stained by a modified Papanicolaou method. To determine interobserver reproducibility, the differential cell counts of two blinded cytopathologists were compared. A cytodiagnosis of AR was made when the urine sample contained < 55% neutrophils and > 20% lymphocytes. To determine the validity of the cytology, the result was compared to the histologic and clinical diagnoses. Biopsies were obtained one hour following vascular anastomosis and at the time of graft dysfunction and were scored by two blinded pathologists according to the Banff classification. The clinical diagnosis was determined by a retrospective review conducted by four blinded clinicians. RESULTS: The interoperator reading of urine cytology was more reproducible than histology, with kappa values of 0.40 +/- 0.15 (SE) and 0.21 +/- 0.10 (SE), respectively. Urine cytology was accurate for the diagnosis of AR, with a sensitivity of 80% and a specificity of 96% as compared to the clinical and histologic findings. CONCLUSION: Our observations support the claim that urine cytology is useful for diagnosing AR. PMID- 9390134 TI - Urine cytology and the diagnosis of renal allograft rejection. II. Studies using immunostaining. AB - BACKGROUND: Urine immunocytology may provide a noninvasive method of investigating the antigens expressed by renal tubular cells. In previous investigations of patients with acute renal allograft rejection (AR), we showed that the adhesion molecule ICAM-1 is expressed by voided tubular cells. The up regulation of ICAM-1, in turn, may be due to high circulating levels of interferon-gamma and/or TNF-alpha. We investigated the regulation of receptors for these cytokines and found a correlation between their expression and clinical events. STUDY DESIGN: For 10 patients who received transplants consecutively, freshly voided aliquots of urine were obtained on each hospital day and on each outpatient visit for a mean of 52.8 +/- 26.2 (SD) days. After cytocentrifugation, the samples were prepared by the avidin-biotin-immunoperoxidase technique in order to detect the presence or absence of ICAM-1, interferon-gamma receptor and TNF-alpha receptor (p 80) on the tubular cells. RESULTS: In nonrejecting patients, the tubular cells expressed the interferon-gamma receptor but not ICAM 1 or the TNF-alpha receptor. In patients with AR, the pattern was different. The tubular cells expressed ICAM-1 and the TNF-alpha receptor but not the interferon gamma receptor. CONCLUSION: Urine immunocytochemistry may be useful to demonstrate the expression of cytokine receptors by renal epithelia. PMID- 9390135 TI - Stromal fragments in invasive carcinoma. Source of diagnostic difficulty in aspiration cytology. AB - OBJECTIVE: To analyze three cases of stromal fragments in invasive carcinoma that created diagnostic difficulty in aspiration cytology. STUDY DESIGN: A retrospective review of fine needle aspiration cytology (FNAC) smears of a breast tumor, scalp tumor and neck mass. Cytomorphologic features of all the smears were reviewed after histology became available. RESULTS: FNAC smears revealed a biphasic pattern: a carcinomatous component and a stromal component that was either discrete or in close apposition to the carcinoma. The cytopathologist had suggested the diagnosis of a biphasic tumor in each case--phyllodes, malignant skin adnexal and salivary gland tumor. Histopathology revealed an invasive carcinoma with altered stroma in the first two cases and metastatic lymph node with perinodal soft tissue extension in the third case. CONCLUSION: Stromal changes in response to infiltrating carcinoma are well documented in surgical pathology. However, these may also be encountered in FNAC smears. The above cases stress the importance of recognizing stromal fragments in aspiration cytology in order to avoid diagnostic errors. PMID- 9390136 TI - Diagnosis of occult thyroid carcinoma by thyroid ultrasonography with fine needle aspiration cytology. AB - OBJECTIVE: To assess the role of ultrasonography and fine needle aspiration cytology (FNAC) in preoperative diagnosis of patients with occult thyroid carcinoma (OTC). STUDY DESIGN: Data on 768 thyroid carcinoma patients receiving primary treatment at Chang Gung Medical Center were retrospectively reviewed. Of these patients, 97 had OTC. To detect small thyroid nodules early and define the characteristics of clinically palpable nodules, thyroid ultrasonography with FNAC were performed on 67 histopathologically proven OTC patients. Analysis for diagnostic value was done for ultrasonography and FNAC. RESULTS: In the 67 patients receiving ultrasonography with FNAC, 23 were preoperatively diagnosed as having papillary thyroid carcinoma and 1 as having follicular carcinoma. The tumor size of these 24 preoperative FNAC-proven OTC was 0.81 +/- 0.23 cm (mean +/ SD). In the remaining patients, 10 presented pictures suspicious for malignancy, with a mean tumor size 0.63 +/- 0.24 cm, and 33 (49.3%) were diagnosed as having benign thyroid lesions in preoperative FNAC. The tumor size in these 33 lesions was 0.58 +/- 0.24 cm. Fifty-seven of the 67 OTC patients received frozen sections. Thirty-eight papillary thyroid carcinomas and four follicular carcinomas were correctly diagnosed on frozen sections. CONCLUSION: Although the rate is not high, high-resolution ultrasonography and FNAC is the best approach to preoperative diagnosis for OTC patients today. PMID- 9390137 TI - Calretinin. A selective marker of normal and neoplastic mesothelial cells in serous effusions. AB - OBJECTIVE: To document that a polyclonal antiserum to calretinin, a 29-kd calcium binding protein, consistently decorates normal and tumor mesothelial cells in cytologic preparations. STUDY DESIGN: Thirty-three archival cytologic specimens from eight patients with histologically confirmed malignant mesothelioma and 13 from patients with metastatic serous effusions were destained and then immunostained with anticalretinin antiserum. For investigation of cell suspensions, four pleural fluids were incubated with anticalretinin antiserum. After cytocentrifugation the specimens were stained in accordance with the alkaline phosphatase anti-alkaline phosphatase (APAAP) method. For electron microscopic examination the cell suspensions were then incubated with gold labeled antirabbit antibody. RESULTS: The diagnostic sensitivity of this new immunocytochemical approach reached 100% for the eight malignant mesotheliomas investigated. Only 3 of the 13 adenocarcinomas metastatic to the serous membranes included in this study were weakly reactive, accounting for 81% specificity. Binding of anticalretinin antiserum to living mesothelial cells was consistently documented in all four cases investigated. CONCLUSION: Calretinin is a very useful marker for positive identification of normal and tumor mesothelial cells in serous effusions. PMID- 9390138 TI - Correlation of the ratio of CD4+/CD8+ cells in lymph node fine needle aspiration biopsies with HIV clinical status. A preliminary study. AB - OBJECTIVE: To test the hypothesis that lymph node (LN) fine needle aspiration biopsy (FNAB) may provide reliable measures of human immunodeficiency virus (HIV) disease status. STUDY DESIGN: HIV+ participants in this study had persistent generalized lymphadenopathy without clinical evidence of lymphoma or nodal infections due to organisms other than HIV. Seven males and five females ranging in age from 23 to 55 and at HIV Centers for Disease Control (CDC) stages A2-C3 were enrolled in this study. From each participant, LN and blood samples were submitted for cytologic examination and flow cytometric analysis of lymphocyte subsets. Flow cytometry measures included T, B, CD4+, CD8+ and natural killer (NK) cells. The percentages of T, B and NK cells in LN and blood samples were different and reflected the expected distribution of these cell types in the respective tissues. RESULTS: The percentages of CD4+ and CD8+ cells in blood and LN were different, but this variation was not statistically significant. In contrast, the ratio of CD4+/CD8+ cells in LN and blood was different and statistically significant (P < .001) for patients in CDC categories A2-B2 but not different for categories B3-C3. More important, there was a significant (r = .76) correlation between the ratio of CD4+/CD8+ cells in LN with CDC stage. CONCLUSION: FNAB, in combination with flow cytometry, may prove to be an important tool in HIV clinical staging. However, further assessment, including clinical follow-up and participation of additional patients, is necessary and currently under way. PMID- 9390139 TI - Concurrent fluorescence in situ hybridization and immunocytochemistry for the detection of chromosome aberrations in exfoliated bronchial epithelial cells. AB - OBJECTIVE: A procedure was developed to allow concurrent detection of chromosome aberrations and identification of bronchial epithelial cells. STUDY DESIGN: Fluorescence in situ hybridization for chromosome 7 and immunocytochemistry for cytokeratin were performed on exfoliated bronchial epithelial cells in a sputum sample from a cancer patient. RESULTS: The Spectrum Orange-labeled alpha satellite probe for chromosome 7 produced red fluorescence, nuclei were counterstained with 4,6-diamidino-2-phenylindole (blue), and cytokeratin was visualized using a fluorescein isothiocyanate (FITC)-conjugated secondary antibody (green). CONCLUSION: This procedure allowed the rapid identification of airway epithelial cells with numerical chromosome aberrations in this sample. Ultimately, this procedure could increase the sensitivity and specificity of sputum cytology as a laboratory diagnostic tool for the early detection of lung cancer. PMID- 9390140 TI - Saccomanno smear slides and Megafunnel slides for sputum specimens. A comparison. AB - OBJECTIVE: To compare Megafunnel slides to standard Saccomanno smear slides of sputum specimens and evaluate the use of Megafunnel slides for retrospective studies. STUDY DESIGN: Papanicolaou-stained Saccomanno smear and Megafunnel slides (Shandon Lipshaw, Inc., Shandon Inc., Pittsburgh, Pennsylvania, U.S.A.) of 65 clinical sputum specimens from 51 patients were compared for cellular morphology, staining, background and cytologic diagnosis. Recovery of diagnostic cells was quantitated using 10 of these specimens. Megafunnel slides prepared from the clinical sputum samples were immunocytochemically stained. Diagnostic cells were quantitated both before removal from 64 archived Saccomanno smear slides and after placement of these cells onto 238 Megafunnel slides. RESULTS: Saccomanno smear slides and Megafunnel slides of clinical specimens were similar in morphology, background, staining, diagnosis and cell recovery. Megafunnel slides were superior for multiple immunocytochemical stains. The production of multiple Megafunnel slides from archival smear slides provided a method of performing numerous retrospective studies. CONCLUSION: Megafunnel slides compared favorably to Saccomanno smear slides in the quality of specimens but are more expensive and labor intensive to prepare. However, the reduction in screening time by cytotechnologists may be advantageous. Additionally, their potential use for immunocytochemistry, fluorescence in situ hybridization, or other special clinical and research analyses is very promising. PMID- 9390141 TI - Taking a satisfactory cervical cytologic smear. Is it really an easy procedure? AB - OBJECTIVE: To evaluate the correlation between the experience of the cervical cytologic smear provider and the quality of the smears in terms of the percentages of satisfactory smears and contribution of the various factors affecting the smear's adequacy. STUDY DESIGN: A newly available quality control system was used to evaluate the adequacy of 4,000 smears. RESULTS: Of 4,000 smears, 660 (16.5%) were classified as "satisfactory but limited" or "unsatisfactory." Technical factors contributed 1.0%, while 15.5% were due to sampling factors, considered human errors. Thus, human error accounted for 90% of the total number of unsatisfactory smears. CONCLUSION: Greater experience with smear sampling is associated with fewer unsatisfactory smears. The results correlate directly with the total number of smears taken annually. Sampling skill improves in steps, with improvement limited beyond a certain point. PMID- 9390142 TI - Application of the CytoRich monolayer preparation system for cervical cytology. A prelude to automated primary screening. AB - OBJECTIVE: To compare AutoCyte's CytoRich monolayer preparation method with the conventional cervical cytology smear method as a reliable alternative for cervical cytology screening. STUDY DESIGN: After conventional smears were prepared, sample collection devices were placed in vials containing CytoRich cell preservative fluid. The residual sample material was rinsed into the vials and prepared on glass slides by the CytoRich system. CytoRich monolayer preparations were then compared microscopically with matched conventional smears in a blind assessment in order to confirm whether diagnostic cells were present on the glass slides when the CytoRich method was used. RESULTS: A total of 2,000 samples, including cases with low grade squamous intraepithelial lesion (SIL), high grade SIL and cancer were studied. Of 58 cases diagnosed as SIL from either the conventional or CytoRich preparation, 46 cases were in agreement. The overall agreement, including negatives and abnormal cases greater than low grade SIL, was 98.8%. CONCLUSION: Dysplastic and cancer cells were detected satisfactorily on the 13-mm-diameter CytoRich monolayers. These preparations are not only satisfactory for rapid manual microscopic examination but also are applicable for evaluation using automated screening systems. PMID- 9390143 TI - Primary malignant melanoma of the conjunctiva of the upper eyelid. A case report. AB - BACKGROUND: Malignant melanoma of the conjunctiva is rare. The nomenclature and clinical and pathologic features of cutaneous and conjunctival melanomas are different. CASE: A 62-year-old male presented with a history of slight bleeding of the upper conjunctiva for the previous six months. On clinical examination the ophthalmologist observed a smooth, partly nodular, pigmented lesion on the conjunctiva under the left eyelid, 1.5 cm in diameter. Fine needle aspiration (FNA) biopsy of the mass showed tumor cells dispersed as single cells with eccentric, round nuclei; coarsely granular chromatin; prominent nucleoli; and dense cytoplasm with occasional brownish pigmentation as well as small aggregates of spindle-shaped neoplastic cells with hyperchromatic nuclei and no cytoplasmic pigment. CONCLUSION: FNA cytology is a simple and efficient method of making the diagnosis of malignant melanoma in conjunctival masses. Careful correlation with the clinical history and histologic findings is often necessary for confirmation of the diagnosis. PMID- 9390144 TI - Fine needle aspiration cytology of Langerhans cell histiocytosis confined to lymph nodes. A case report. AB - BACKGROUND: Lymph node involvement in Langerhans cell (LC) histiocytosis (LCH) can be seen as a component of the systemic form, or it may be the initial and sometimes exclusive manifestation of the disease. Descriptions of patients with LCH whose disease is confined to lymph nodes are rare. CASE: We present a case of LCH confined to lymph nodes initially diagnosed by fine needle aspiration (FNA) cytology in a 43-year-old male. The cytologic findings in LCH included high cellularity, isolated LCs with prominent nuclear indentations and grooves, multinucleate giant cells, eosinophils and lymphocytes. Confirmation of LCH was obtained by positive S-100 protein immunohistochemical staining and the demonstration of Birbeck granules on electron microscopy. CONCLUSION: The presence of LCs with prominent nuclear indentations and grooves is characteristic of LCH confined to lymph nodes and serves as a key point in suggesting the diagnosis of LCH. PMID- 9390145 TI - Mucor pyelonephritis. Report of a case diagnosed by urine cytology, with diagnostic considerations in the workup of funguria. AB - BACKGROUND: Isolated renal mucormycosis is an uncommon kidney infection affecting patients with underlying systemic diseases and intravenous (IV) drug abuse. We report a unique case in the cytologic literature in which urine cytology provided insight into the diagnosis, renal mucormycosis. CASE: The patient, a diabetic and IV drug abuser, presented with complaints of left flank pain, fever and dysuria. All urine cultures were negative. A computed tomography (CT) scan showed changes consistent with left acute pyelonephritis, and the patient was treated for a presumed diagnosis of bacterial pyelonephritis. Late in the hospital stay, the cytology laboratory diagnosed Mucor in a single urine specimen, but the patient had already been discharged. The patient was never treated for funguria, only to present again with left flank pain 13 months later. An abdominopelvic CT scan showed progression to left chronic pyelonephritis. The patient, however, left the hospital against medical advice before any further workup could be completed. CONCLUSION: Renal mucormycosis should be considered part of the differential diagnosis in patients with underlying diseases or IV drug abuse who present with symptoms of acute pyelonephritis. The differential diagnosis of Mucor funguria should also include fungal ball in the renal pelvis or urinary bladder and fungal cystitis. PMID- 9390146 TI - Fine needle aspiration cytology of gastric epithelioid leiomyosarcoma metastasized to the liver. A case report. AB - BACKGROUND: There have been only a few reports on fine needle aspiration (FNA) cytology of epithelioid leiomyosarcoma, especially of the stomach, and a summary of the cytologic findings in this tumor is needed. CASE: A case of epithelioid leiomyosarcoma of the stomach metastasized to the liver and was composed cytologically of peculiar binucleated cells. CONCLUSION: Similar findings in most cases are that the cells are round or polygonal, with eccentrically located nuclei. The most variable findings relate to the texture of the cytoplasm, which varies from granular to dense to vacuolar. FNA cytology of epithelioid leiomyosarcoma can show cells that are mononuclear, binucleated or multinucleated, with eccentric nuclei and dense to vacuolar cytoplasm, with the variations probably depending on fixation status. PMID- 9390147 TI - Granular cell tumor of the breast in a male. A case report. AB - BACKGROUND: Granular cell tumor (GCT) is a rare tumor of the soft tissues; the most common site of occurrence is the tongue. The diagnosis of GCT is fairly straightforward on both fine needle aspiration cytology and histopathology. Occasionally the tumor's presence in an unusual site may create confusion with carcinoma, especially when it occurs in the breast, where carcinoma is more common. CASE: A case of GCT occurred in the breast of a male with simultaneous detection of the same type of tumor in the soft tissues of the upper arm and back. The diagnosis was made on a fine needle aspirate of the breast mass, which demonstrated characteristic diastase-resistant, periodic acid-Schiff-positive intracytoplasmic granules. CONCLUSION: The breast, especially in males, is an unusual location for GCT. Clinicians and pathologists must be alert, therefore, to its existence and its inclusion in the differential diagnosis of tumors of the breast. PMID- 9390148 TI - Pigmented villonodular synovitis. Report of a case with diagnostic synovial fluid cytologic features. AB - BACKGROUND: The identification of neoplastic cells in synovial fluid is an uncommon occurrence and most often is related to extension of extraarticular tumor into the joint space than to primary neoplasm arising in the joint. CASE: In this report the cytologic features of synovial fluid obtained from the right knee of an 18-year-old male with biopsy-proven pigmented villonodular synovitis are described and compared with the features of the concurrent surgical specimen. CONCLUSION: The cytologic features of pigmented villonodular synovitis in synovial fluid include abundant mononuclear histiocytic cells occurring singly and in papillary clusters, hemosiderin within histiocytes and few multinucleated giant cells. PMID- 9390149 TI - Fine needle aspiration biopsy of progressive multifocal leukoencephalopathy in a patient with AIDS. A case report. AB - BACKGROUND: Progressive multifocal leukoencephalopathy (PML) is one of the most common opportunistic infections, with a range of 4-7% in acquired immunodeficiency syndrome (AIDS) patients. Clinical diagnosis is often difficult, and the specific pathologic agent requires cytologic and pathologic confirmation. CASE: A 38-year-old, Haitian male was admitted with a new-onset seizure disorder. On computed tomography (CT), there were right frontoparietal cortex, right external capsule and right basal ganglia lucencies. Fine needle aspiration biopsy (FNAB) of the radiolucent area revealed foci of white matter demyelination and a few eosinophilic inclusions in oligodendrocytes plus abnormal giant astrocytes. Ultrastructurally, JC virions were observed in the nuclei and cytoplasm of the oligodendrocytes. CONCLUSION: Diagnostic cranial CT-guided FNAB, with cytologic and histologic studies, is extremely valuable in evaluating the nature of central nervous system demyelinated and space-occupying lesions in AIDS. PMID- 9390150 TI - Subareolar abscess of the breast in a male. A report of two cases with fine needle aspiration cytology diagnosis. AB - BACKGROUND: Fine needle aspiration (FNA) is a well-established method for the diagnosis of breast diseases in males, but very little attention has been focused on inflammatory lesions. Previous reports do not mention subareolar abscess (SA), which is a distinct entity in breasts of females. CASES: Two adult males presented for FNA of recurrent breast masses located near the nipple. Smears from both patients showed the typical cytologic features of SA. CONCLUSION: Recognition of this troublesome and rare condition on FNA cytology from the breasts in males will prevent recurrences since SA is not cured by excisional biopsy. PMID- 9390151 TI - Solid and papillary epithelial neoplasm of the pancreas. Report of a case with diagnosis by fine needle aspiration cytology. AB - BACKGROUND: Solid and papillary epithelial neoplasm of the pancreas is a distinct clinicopathologic entity. It has a benign clinical course, and surgical resection can be curative. CASE: A 17-year-old female presented with a mass measuring about 12 cm in the epigastrium and left hypochondrium. Fine needle aspiration cytology (FNAC) showed papillae with a central fibrovascular core and lined with many layers of bland-appearing cells exhibiting nuclear grooving. These features, along with ultrasound and computed tomographic findings, led to an accurate preoperative diagnosis. CONCLUSION: In the setting of typical clinical and radiologic findings, it is possible to make a correct preoperative diagnosis by FNAC. Doing so has important implications for management. PMID- 9390152 TI - Aspiration cytology of signet-ring cell lymphoma. A case report. AB - BACKGROUND: Signet-ring cell lymphoma is a rare subtype of non-Hodgkin's lymphoma, composed of vacuolated cells with a signet-ring cell appearance. We found only two cases that had been reported as diagnosed by fine needle aspiration biopsy. CASE: A 61-year-old female had signet-ring cell lymphoma diagnosed by computed tomography-guided aspiration biopsy. Smears of aspirates from her retroperitoneal mass contained a population of small to medium-sized, angular lymphoid cells; lymphoglandular bodies; and an abundance of signet-ring cells. The signet-ring cells were negative for cytokeratin and positive for leukocyte common antigen and CD20, a B-cell marker. Monoclonality for lambda light chain determinant was noted, and a diagnosis of signet-ring cell lymphoma of the B-cell type was made. A core biopsy specimen confirmed the diagnosis. CONCLUSION: Signet-ring cell lymphoma should always be considered in the differential diagnosis of tumors composed of signet-ring cells. PMID- 9390153 TI - Pulmonary malakoplakia diagnosed by fine needle aspiration. A case report. AB - BACKGROUND: Pulmonary malakoplakia is an uncommon disorder, with 24 previously reported cases, only 4 of which were diagnosed by bronchial washings, bronchial brushings or aspiration cytology. We report a case that was diagnosed initially by computed tomography (CT)-guided fine needle aspiration (FNA) cytology. CASE: A 56-year-old male with follicular small cleaved cell lymphoma had a 10-cm left lower lobe mass compressing the main bronchus to that lobe. A transthoracic, CT guided FNA specimen consisted predominantly of foamy macrophages, many of which contained typical Michaelis-Gutmann bodies. Microbiologic cultures identified Rhodococcus equi. A subsequent transbronchial biopsy and left pneumonectomy specimen confirmed the cytologic diagnosis. CONCLUSION: Pulmonary malakoplakia associated with R equi pneumonia is a rare lesion that is essentially limited to immunocompromised hosts. Awareness of the FNA cytomorphology of this lesion permits resolution of the typical clinical differential diagnosis of pulmonary masses in the immunocompromised host and can facilitate treatment. PMID- 9390154 TI - Thymic carcinoid. Report of a case with diagnosis by fine needle aspiration biopsy. AB - BACKGROUND: Fine needle aspiration biopsy (FNAB) affords a less expensive, less morbid approach to masses within the complex anatomy of the mediastinum as opposed to surgical biopsy. Given the current state of computed tomography guidance and the available cell block preparations and ancillary studies, definitive diagnosis of mediastinal tumors is possible. CASE: A 19-year-old male presented with weight loss and muscle weakness. Computed tomography revealed an anterior superior mediastinal mass with attachment to the posterior sternum and anterior aorta. FNAB yielded hyperchromatic cells with densely clumped chromatin and prominent nucleoli. These were present as single cells and clusters. Cell block preparations were studied with immunoperoxidase methods and were strongly positive for chromogranin and glucagon, supporting the diagnosis of carcinoid tumor. Surgical excision yielded a 7-cm, unencapsulated, red-brown tumor with medium-sized cells with oval to round nuclei, scant and granular cytoplasm and coarse "salt and pepper" chromatin with prominent nucleoli. The cells were arranged in islands and bands and were associated with prominent capillaries and dense, collagenous septae. Immunoperoxidase and electron microscopy demonstrated numerous intracytoplasmic, nonspecific neurosecretory granules and positivity for somatostatin, synaptophysin, cytokeratin and chromogranin. CONCLUSION: FNAB affords an accurate and timely diagnosis of an anterior mediastinal tumor without the necessity for open biopsy and also offers accurate surgical planning and decreased morbidity. PMID- 9390155 TI - Fine needle aspiration diagnosis of a subcutaneous abscess from Enterobius vermicularis infestation. A case report. AB - BACKGROUND: Extraintestinal infestation by Enterobius vermicularis is uncommon. It has been reported to occur in the peritoneal cavity, ovary, fallopian tube, endometrium, lung, liver and urinary tract. CASE REPORT: Fine needle aspiration diagnosis was made in a case of enterobiasis presenting with a subcutaneous abscess in the natal cleft. Eggs, as well as fragments of cuticle of the adult worm, were found; the morphology of both was best visualized in Papanicolaou stained smears. Polarizing microscopy highlighted the equally spaced parallel grooves of the cuticle. CONCLUSION: Fine needle aspiration cytology of subcutaneous abscesses due to enterobiasis can be diagnostic when eggs, or eggs with cuticle, are identified in a suppurative or granulomatous inflammation. PMID- 9390156 TI - Detecting false negative smears. PMID- 9390157 TI - Fine needle aspiration cytology of injection-site pseudotumors. PMID- 9390158 TI - Myoepithelioma of the soft palate. PMID- 9390159 TI - Cytologic diagnosis of hepatoblastoma by fine needle aspiration biopsy cytology. PMID- 9390160 TI - Recognition of rectal glandular cells in vaginal smears. PMID- 9390161 TI - Use of water-soluble gel in obtaining the cervical cytologic smear. PMID- 9390162 TI - Cytologic diagnosis of Bancroft's filariasis presenting as generalized lymphadenopathy. PMID- 9390163 TI - ASCUS: a misnomer. PMID- 9390164 TI - Postmastectomy angiosarcoma (Stewart-Treves syndrome) PMID- 9390165 TI - Human papillomavirus detection in archival Papanicolaou-stained cervical smears by in situ polymerase chain reaction. PMID- 9390166 TI - Reactive lymphoid hyperplasia presenting as a palpable nodule in the breast. PMID- 9390167 TI - Trichosporon detected on bile cytology. PMID- 9390168 TI - Isolation of carboxylester lipase (CEL) isoenzymes from Candida rugosa and identification of the corresponding genes. AB - The yeast Candida rugosa produces extracellular lipases which are widely used for industrial purposes. A commercial lipase preparation from this yeast can be separated into several isoenzymes which differ in carbohydrate content, isolelectric point, substrate specificity, and primary sequence. We have here purified and characterized three lipases, which also hydrolyze p-nitrophenyl esters, from a commercial preparation of this yeast. These three carboxylester lipases (CELs) elute differently on hydrophobic interaction chromatography, and have different carbohydrate contents and substrate specificities. Sequence analysis of their amino termini and peptides generated by LysC treatment showed that CEL-1 and CEL-3 probably have identical primary structure while CEL-2 was proven to be a different enzyme. Sequence comparison showed that both CEL-1 and CEL-3 are products of the LIP1 gene and that CEL-2 is the gene product of LIP2, cloned by Longhi et al. (Biochim. Biophys. Acta 1131, 227-232, 1992). PMID- 9390169 TI - Mechanisms for metabolism of ethanol to 1-hydroxyethyl radicals in rat liver microsomes. AB - Experiments have been designed to reevaluate mechanisms for metabolism of ethanol to 1-hydroxyethyl radicals (HER) in rat liver microsomes. The variables tested include addition of azide, catalase, superoxide dismutase, and deferoxamine, or use of phosphate or Tris buffers. The results indicate that several mechanisms of HER formation are possible, depending on the experimental conditions used to study this process. In the presence of phosphate buffer, which has been used extensively in spite of its ability to chelate iron, HER formation is quite sensitive to changes in hydrogen peroxide availability. These results suggest that Fenton-type reactions produced the oxidizing intermediate responsible for conversion of ethanol to a free radical in phosphate buffer. However, in Tris buffer, HER formation was inhibited markedly by addition of superoxide dismutase, whereas catalase or azide had little effect. These data indicate that the apparent mechanism of radical formation may be influenced by the choice of buffer used. HER formation was almost abolished by the combination of superoxide dismutase and deferoxamine in both buffers, suggesting little enzymatic HER formation by the cytochrome P450 enzymes. When changes in HER formation were compared with rates of ethanol oxidation, it was inferred that 25 to 50% of the acetaldehyde formed during microsomal ethanol oxidation under different experimental conditions could arise via the HER intermediate. PMID- 9390170 TI - Purification and characterization of a rat liver bile acid coenzyme A ligase from rat liver microsomes. AB - In the present study, using the C24 bile acid chenodeoxycholic acid as substrate, rat liver bile acid CoA ligase activity (rBAL) was purified 200-fold from detergent-solubilized microsomes using a combination of Q-Sepharose anion exchange, hydroxyapatite, and CM-Sepharose chromatography. Purified rBAL had a molecular weight of 65 kDa by SDS-PAGE analysis. Gel filtration of purified rBAL indicated that rBAL activity forms a complex with other proteins with an apparent aggregate molecular weight of 243 kDa. A monoclonal antibody raised against the 65-kDa protein and covalently coupled to 6B-Sepharose completely absorbed rBAL activity from a semipurified preparation of rat liver microsomes. Western blot analysis confirmed the elution of the 65-kDa protein from the affinity phase at low pH. Optimum rBAL activity was found at pH 8.5, and activity was dependent on the divalent cation Mg2+. In the presence of 50 microM CoA and 2.5 mM MgCl2, kinetic analysis revealed that the apparent K(m)s of ATP and chenodeoxycholic acid of the purified enzyme were 548 +/- 247 and 18.0 +/- 6.2 microM, respectively, and the apparent Vmax was 9.53 +/- 2.0 nmol min-1 mg protein-1. The formation of chenodeoxycholyl-CoA by rBAL was strongly inhibited by hydrophobic bile acids (the C24 monohydroxy bile acid lithocholic acid and 3 alpha,7 alpha,12 alpha-trihydroxy-5 beta-cholestanoic acid, the C27 homolog of cholic acid), but only weakly by cholic acid. Chenodeoxycholyl-CoA and 3 alpha,7 alpha,12 alpha trihydroxy-5 beta-cholestan-27-oyl-CoA were confirmed as reaction products of purified rBAL by HPLC-electrospray ionization mass spectrometry. PMID- 9390171 TI - Cancer-preventive selenocompounds induce a specific redox modification of cysteine-rich regions in Ca(2+)-dependent isoenzymes of protein kinase C. AB - Since protein kinase C (PKC) serves as a receptor for phorbol ester type tumor promoters and oxidants and has unique redox-active cysteine-rich regions, we have determined whether various chemopreventive selenocompounds could affect this enzyme. At lower concentrations, selenite decreased the kinase activity (IC50 = 0.5 microM), while at higher concentrations it decreased phorbol ester binding. However, when the catalytic and regulatory domains of PKC were separated by proteolysis, the catalytic domain retained its sensitivity to selenite, while the regulatory domain lost its sensitivity. Cysteine residues were quantitated in PKC modified with selenite by using 5,5'-dithiobis(2-nitrobenzoic acid) and also by using 2-nitro-5-thiosulfobenzoic acid after sulfitolysis. At lower concentrations, selenite induced a modification of four cysteine residues resulting in the formation of two disulfides, while at higher concentrations it induced a modification of seven to eight cysteine residues resulting in the formation of three to four disulfides. Contrary to selenite, selenocystine and selenodiglutathione (GSSeSG) readily inactivated the kinase activity, but not the phorbol ester binding. These two agents induced a two-stage modification of PKC; a limited modification at low concentrations leads to a loss of affinity for ATP, while an excessive modification at high concentrations leads to a loss of Vmax. Selenocystine and GSSeSG were 100,000-fold more potent than GSSG in inactivating PKC. The isoenzymes alpha, beta, and gamma exhibited an identical susceptibility to these selenocompounds. These results suggested that the cysteine residues present within the catalytic domain of these isoenzymes, although apart in the sequence, may be clustered in the tertiary structure to react with selenite, as well as may be in close proximity to some of the cysteines in the regulatory domain. Selenite did not affect protein kinase A, whereas GSSeSG and selenocystine inactivated the catalytic subunit after dissociation from the regulatory subunit at concentrations 100- and 800-fold, respectively, higher than that required for PKC inactivation. All three selenocompounds did not affect the activities of phosphorylase kinase and protein phosphatase 2A. Taken together, these results suggest that the accessible redox-active cysteine residues present in the PKC catalytic domain can react with certain specificity with redox-active selenocompounds such as selenite, selenocystine, and GSSeSG relative to other protein kinases tested. PMID- 9390172 TI - Selenocompounds induce a redox modulation of protein kinase C in the cell, compartmentally independent from cytosolic glutathione: its role in inhibition of tumor promotion. AB - Since selenite and other redox-active selenocompounds can modify protein kinase C (PKC) in the test tube, we have determined whether or not this redox regulation occurs inside the cell despite having high concentrations of GSH and the role of this regulation in the inhibition of tumor promotion. By using phorbol ester promoted JB6 epidermal cell transformation assay, the concentrations of selenite, selenocystine, and selenodiglutathione which are optimal for chemopreventive activity were determined. At such concentrations (0.5 to 2 microM) in the cells treated with these agents, only a slight but transient decrease in PKC activity was observed when measured with a low (5 microM), but not with a high (100 microM) concentration of ATP. However, when the cells were serum starved or pretreated with 2-deoxyglucose, there was a pronounced but transient inactivation of PKC when assayed with both low and high concentrations of ATP. The inactivation was reversed in the cell by an endogenous mechanism or by treatment with thiol agents in the test tube. In spite of a substantial (90%) depletion of GSH in the cells by pretreatment with buthionine sulfoximine, there was no further increase in the redox modification of PKC by selenite as well as no change in the inhibitory effect of selenite on the phorbol ester-stimulated induction of ornithine decarboxylase, which is an intermediate marker related to cell transformation. While GSH is known to influence certain actions of selenium, it may not be required to mediate the effects of selenite tested in this study. The water-soluble cytosolic GSH did not interfere with the redox modification of PKC probably due to the shielding of the cysteine-rich region of the enzyme by a weak hydrophobic association with the membrane. Due to the presence of cofactors in the crude cell extracts, PKC was more sensitive to selenite than in the purified form and was inactivated by low concentrations of selenite (IC50 = 0.05 microM). This modification was reversed by thiol agents as well as by NADPH. A protein disulfide reductase, which can regenerate PKC, was present in the homogenate. Conceivably, selenite and other selenocompounds induce a redox modification of cellular PKC, compartmentally independent from the cytosolic GSH, but intimately connected to a NADPH-dependent reductase system, to mediate, at least in part, some of the cancer-preventive actions. PMID- 9390173 TI - Dependence of salt concentration on glycosaminoglycan-lysozyme interactions in cartilage. AB - The cationic protein, lysozyme, has an extracellular distribution in cartilage but its precise role in this tissue has not yet been established. This study describes the dependence of salt concentration on the binding properties of lysozyme isoforms of different cationic charges, isolated from bovine cartilage, to the two major and structurally similar glycosaminoglycans of cartilage, i.e., chondroitin sulfate and hyaluronan. The binding of most cartilage lysozyme isoforms and hen egg-white lysozyme (control) to chondroitin sulfate and hyaluronan linked to agarose supports displayed optimal levels at approximately 20 and 5-10 mM salt, respectively, but decreased at both lower and higher salt concentrations indicating the electrostatic nature of the interactions. However, optimal binding of the most cationic lysozyme isoform to chondroitin sulfate occurred at 60 mM salt, with significant binding remaining at 150 mM. This isoform also showed binding to hyaluronan up to 60 mM salt, while for the other isoforms binding was observed only up to 150 and 40 mM salt for chondroitin sulfate and hyaluronan, respectively. The low salt concentrations at which these interactions occur are likely to exist in cartilage as shown from equilibrium dialysis studies performed using solutions of chondroitin sulfate (up to 10%, a concentration likely to occur in cartilage). From Scatchard analysis, the affinity of binding of all lysozymes to chondroitin sulfate was similar (Kd = 10( 6) M) and slightly lower than their binding to hyaluronan (Kd = 10(-7) M) of similar molecular mass. PMID- 9390174 TI - Microsomal alcohol oxygenase: purification and characterization of a cytochrome P450 responsible for oxidation of 7-hydroxy-delta 8-tetrahydrocannabinol to 7-oxo delta 8-tetrahydrocannabinol in guinea pig liver. AB - Guinea pig hepatic enzyme, microsomal alcohol oxygenase, was able to oxidize both 7 alpha- and 7 beta-hydroxy-delta 8-tetrahydrocannabinol (7 alpha- and 7 beta hydroxy-delta 8-THC) to 7-oxo-delta 8-THC. A cytochrome P450, named P450GPF-B, which mediates this oxidative metabolism was purified from hepatic microsomes of untreated female guinea pigs. The purified enzyme showed a single protein band of molecular mass 50,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The NH2-terminal amino acid sequence of P450GPF-B is highly homologous with those of several cytochrome P450s belonging to the CYP3A subfamily. 18O derived from atmospheric oxygen was incorporated into 31 and 6%, respectively, of 7-oxo-delta 8-THC formed from 7 alpha- and 7 beta-hydroxy-delta 8-THC when the substrates were incubated with P450GPF-B under 18O2. The antibody against P450GPF-B significantly suppressed the oxidative activities of 7 alpha- and 7 beta-hydroxy-delta 8-THC to 7-oxo-delta 8-THC in hepatic microsomes of guinea pig. These results indicate that P450GPF-B is a major enzyme responsible for the hepatic microsomal alcohol oxygenase activities in the guinea pig. PMID- 9390175 TI - Biochemical and molecular characterization of fumarase from plants: purification and characterization of the enzyme--cloning, sequencing, and expression of the gene. AB - A cDNA EST clone encoding the C-terminal portion of Arabidopsis thaliana fumarase was identified by homology analysis. A fragment of cDNA encoding the N-terminal region of fumarase was amplified from a cDNA library using PCR and cloned. Genomic DNA corresponding to the coding region of fumarase was amplified and cloned. Arabidopsis fumarase was expressed as a chimeric fusion protein and polyclonal antibodies were generated. Fumarase was purified to near-homogeneity (over 600-fold) from etiolated Pisum sativum mitochondria. The identification of fumarase was confirmed by a combination of immunoblot and N-terminal amino acid sequencing. Kinetic analysis of highly purified fumarase yielded a KM(malate) of 0.45 mM and a Vmax(malate) of 650 mumol of fumarate/min/ mg. The pea fumarase was inhibited by the alpha-keto acids pyruvate and alpha-ketoglutarate at low millimolar concentrations. Adenylates were highly inhibitory; the degree of this inhibition was reduced in the presence of Mg2+, suggesting that uncomplexed adenylates are the inhibitory species. PMID- 9390176 TI - Regulatory role of fructose-2,6-bisP on glucose metabolism in frog oocytes: in vivo inhibition of glycogen synthesis. AB - Glycogen synthesis following glucose microinjection in frog oocytes proceeds preferentially by an indirect pathway involving gluconeogenesis from triose compounds. Because of the known regulatory role of fructose-2,6-bisP on glucose utilization in most vertebrate tissues we coinjected [U-14C]glucose and fructose 2,6-bisP into oocytes and observed a marked inhibition of label incorporation into glycogen, with an I50 value of 2 microM, which is similar to the value measured for the in vitro inhibition of oocyte fructose-1,6-bisphosphatase. Other hexoses-bisP were tested: 2,5-anhydromannitol-1,6-bisP was as effective as inhibitor as fructose-2,6-bisP; glucose-1,6-bisP showed some effect although 50% inhibition was obtained at a concentration 10 times higher than with fructose-2,6 bisP; fructose-1,6-bisP had no effect at all. The inhibition pattern for the in vivo glycogen synthesis by these analogs closely matched the one obtained with partially purified oocyte fructose-1,6-bisphosphatase. The intracellular concentration of fructose-2,6-bisP in unperturbed oocytes was found to be between 0.1 and 0.2 microM. Fructose-6-phosphate,2-kinase levels measured in oocyte homogenates were between 0.02 and 0.06 mU per gram of ovary. After 60 min incubation, fructose-2,6-bisP microinjected into the oocytes was almost completely degraded, suggesting that fructose-2,6-bisphosphatase is active in vivo. The results presented in this paper indicate that fructose-2,6-bisP plays an important role in the in vivo regulation of glucose utilization in frog-grown oocytes. PMID- 9390177 TI - Facilitated reduction of beta-amyloid peptide precursor by synthetic oligonucleotides in COS-7 cells expressing a hammerhead ribozyme. AB - Synthetic deoxyoligonucleotides and phosphorothioate-capped oligonucleotides targeted to bases 112-128 of beta-amyloid peptide precursor (beta APP) mRNA were analyzed for their ability to reduce steady-state beta APP in COS-7 cells and in pMEP4-Rz1 cells that express a hammerhead ribozyme targeted to bases beta APP mRNA 133-148. Cells, incubated in the presence of 10 or 25 microM oligonucleotide, remained viable and morphologically identical to untreated control cells for up to 5 days. Antisense deoxyoligonucleotides beta 112C, beta 114C, and beta 116C specifically lowered beta APP in pMEP4-Rz1 cells compared to noncognate and scrambled oligonucleotide controls. The extent of the beta APP reduction did not depend on oligonucleotide length, although it did depend on the presence and proximity of the ribozyme to the oligonucleotides. beta 117N, a phosphorothioate-capped antisense oligonucleotide, also reduced beta APP levels in pMEP4-Rz1 cells; however, in this case the sense control, beta 117S, affected beta APP similarly, indicating that the observed reduction may be nonspecific. These data imply that deoxyoligonucleotides targeted immediately upstream of a ribozyme binding site can work cooperatively in vivo. Localizing the oligonucleotides and ribozyme and substrate targets to the same cellular pools further confirmed this possibility. PMID- 9390178 TI - Identification of the subdomain in the nuclear receptor for the hormonal form of vitamin D3, 1 alpha,25-dihydroxyvitamin D3, vitamin D receptor, that is covalently modified by an affinity labeling reagent. AB - Multiple physiological actions of the hormonal form of vitamin D3, 1 alpha,25 dihydroxyvitamin D3 (1,25(OH)2D3), are mediated by a genomic pathway which is initiated by the highly specific recognition and binding by its cognate receptor (vitamin D receptor, VDR) in the target cells. Thus, knowledge of the three dimensional geometries of the ligand, i.e., 1,25(OH)2D3, and the 1,25(OH)2D3 binding domain of VDR is crucial for a better understanding of diverse physiological roles of this hormone. Recently our laboratory has developed 1 alpha,25-dihydroxyvitamin D3-3 beta-bromoacetate (1,25(OH)2 D3-3-BE) as an affinity labeling reagent for covalently modifying the hormone binding domain of native VDRs from calf thymus and rat osteosarcoma cells and baculovirus-expressed recombinant human VDR (hVDR). In the present report, we report affinity labeling of the hormone binding domain of hVDR, expressed in Escherichia coli as a glutathione S-transferase fusion partner, site-specific cleavage of the affinity labeled VDR with 3-bromo-3-methyl-2-(2-nitrophenylmercapto)- 3H-indole, and identification of the C-terminal subdomain of human VDR containing the putative hormone binding site. PMID- 9390179 TI - Chemosignal transduction in the vomeronasal organ of garter snakes: Ca(2+) dependent regulation of adenylate cyclase. AB - Earthworm shock secretion contains a 20-kDa vomeronasally mediated chemoattractive protein for garter snakes. Both the ligand-receptor binding and the chemoattractivity of ES20 are Ca(2+)-dependent. When ES20 binds to its G protein-coupled receptors in the vomeronasal epithelium, the inositol 1,4,5 trisphosphate (IP3) level is increased, but the level of cAMP is reduced. Furthermore, forskolin-stimulated levels of cAMP are completely blocked by ES20 receptor binding or by Ca2+ alone and the effect of calcium ions can be nullified by EGTA. Previously, we hypothesized that the decrease in cAMP was due to activation of a Ca(2+)-dependent phosphodiesterase. In the present study, we provide evidence that the decrease in cAMP is due mainly to the regulation of adenylate cyclase (AC) activity by Ca2+ or is indirectly mediated by ES20. Results obtained with intact vomeronasal sensory epithelium suggest that the binding of ES20 to its receptors facilitates generation of IP3 which mobilizes intracellularly sequestered Ca2+, resulting in an increase of cystosolic Ca2+. A further increase in cytosolic Ca2+ occurs through Ca2+ influx from extracellular sources. Garter snake vomeronasal AC does not require calmodulin for its activity and shows a biphasic response to increasing concentrations of Ca2+; its activity is modulated both positively and negatively by this bivalent cation. PMID- 9390180 TI - Reconstitution premixes for assays using purified recombinant human cytochrome P450, NADPH-cytochrome P450 reductase, and cytochrome b5. AB - The development of enzyme and buffer premixes for in vitro biotransformation assays is described. The protein premixes contain a mixture of three recombinant human proteins, cytochrome P450 (P450) 3A4, NADPH-P450 reductase, cytochrome b5, and liposomes. The buffer premix contains reagents which, when diluted, provide for optimal metabolic activity with selected P450 3A4 substrates. P450 3A4 premixes were competent in the oxidation of known substrates including testosterone, midazolam, nifedipine, erythromycin, benzphetamine, and amitriptyline. Premixes stored at -80 degrees C for 2 months and those that underwent an additional five freeze/thaw cycles were able to hydroxylate testosterone at turnover rates similar to freshly prepared reconstitution mixes. In addition, premixes stored unfrozen at 4 degrees C for 2 weeks showed no significant loss in the rate of testosterone 6 beta-hydroxylation by P450 3A4. Premixes prepared with and without reduced glutathione, a component which had previously been found to be important for P450 3A4 reactions, were equally efficient at carrying out testosterone hydroxylation under these conditions. Kinetic parameters determined for the metabolism of testosterone, amitriptyline, nifedipine, and benzphetamine using P450 3A4 premixes were compared with human pooled microsomes and insect microsomes prepared from cells infected with a baculovirus containing two cDNA inserts coding for P450 3A4 and NADPH-P450 reductase. Each format gave different Vmax and K(m) values indicating different catalytic efficiencies. Analysis of P450 1A2 premixes which contained different lipid concentrations indicated that Vmax and K(m) could be altered. The availability of human P450 recombinant enzymes and the development of the P450 premixes that remain active after being stored frozen should allow for rapid identification of novel P450 substrates and inhibitors and the development of large-scale screening assays. PMID- 9390181 TI - Apparent equilibrium constants and standard transformed Gibbs energies of biochemical reactions involving carbon dioxide. AB - When carbon dioxide is produced in a biochemical reaction, the expression for the apparent equilibrium constant K' can be written in terms of the partial pressure of carbon dioxide in the gas phase or the total concentration of species containing CO2 in the aqueous phase, referred to here as [TotCO2]. The values of these two apparent equilibrium constants are different because they correspond to different ways of writing the biochemical equations. Their dependencies on pH and ionic strength are also different. The ratio of these two apparent equilibrium constants is equal to the apparent Henry's law constant K'H. This article provides derivations of equations for the calculation of the standard transformed Gibbs energies of formation of TotCO2 and values of the apparent Henry's law constant at various pH levels and ionic strengths. These equations involve the four equilibrium constants interconnecting the five species [CO2(g), CO2(aq), H2CO3, HCO3-, and CO3(2-)] of carbon dioxide. In the literature there are many errors in the treatment of equilibrium data on biochemical reactions involving carbon dioxide, and so several examples are discussed here, including calculation of standard transformed Gibbs energies of formation of reactants. This approach also applies to net reactions, and the net reaction for the oxidation of glucose to carbon dioxide and water is discussed. PMID- 9390182 TI - A model for the explanation of the thermally induced increase of the overall fluorescence in tryptophan-X peptides. AB - In the range of temperature 10-35 degrees C, Trp-X dipeptides show an unusual increase of fluorescence intensity in solution at pH 7. This effect has been recently studied by means of steady-state fluorescence. Although a model involving the deprotonation at the ground state of the zwitterion was proposed, the activation energy for that process could not rule out the involvement of excited state. In order to understand the mechanism of the thermal-induced increase of fluorescence, we present here time-resolved fluorescence experiments on Trp-X and X-Trp dipeptides at different pH and excitation wave-length. The fluorescence lifetimes (tau i) decrease in accord to thermal quenching, with activation energies (Ei) ranging from 4.0 to 6.4 kcal/mol. Under those circumstances where the anomaly was detected the preexponential factors of the longer-lived component increased as well as their fractional fluorescence. This component can be assigned to the anion species. Because of its larger (three- to fourfold) fluorescence quantum yield, compared to that of the corresponding zwitterion, the large increase of the concentration of the anion leads to an increase of the overall emission despite the thermal quenching. Also the decay associated spectra well account for the red shift of the emission fluorescence spectrum, which accompanies the anomaly. Our model well fits the experimental data using a simple equation which combines Van't Hoff and Arrhenius equations; it also explains the presence of the anomalous thermal quenching exclusively in Trp-X dipeptides excited above 290 nm and at pH around neutrality. PMID- 9390183 TI - Molecular cloning and biochemical characterization of bovine spleen myristoyl CoA:protein N-myristoyltransferase. AB - Myristoyl-CoA:protein N-myristoyltransferase (NMT) is an essential eukaryotic enzyme that catalyzes the cotranslational transfer of myristate to the NH2 terminal glycine residue of a number of important proteins of diverse function. We have isolated full-length cDNA encoding bovine spleen NMT (sNMT). The single long open reading frame of 1248 bp of sNMT specifies a protein of 416 amino acids with a predicted mass of 46,686 Da. The protein coding sequence was expressed in Escherichia coli resulting in the production of functionally active 50-kDa NMT. Deletion mutagenesis showed that the C-terminus is essential for activity whereas up to 52 amino acids can be deleted from the N-terminus without affecting the function. One of the N-terminal deletions resulted in threefold higher NMT activity. Genomic Southern analysis indicated the presence of two strong hybridizing bands with three different restriction enzyme digests suggesting the possibility of two copies of the NMT gene in the bovine genome. RNA blot hybridization analysis of total cellular RNA prepared from bovine brain, heart, spleen, lung, liver, kidney, and skeletal muscle probed with bovine sNMT cDNA revealed a single 1.7-kb mRNA. Western blot analysis of various bovine tissues with human NMT peptide antibody indicated a common prominent immunoreactive band with an apparent molecular mass of 48.5-50 kDa in all tissues. Additional immunoreactive bands were observed in brain (84 and 50 kDa), lung (58 kDa), and skeletal muscle (58 kDa). Activity measurements demonstrated that brain contained the highest NMT activity followed by spleen, lung, kidney, heart, skeletal muscle, pancreas, and liver. It appears therefore that mRNA and protein expression do not correlate with NMT activity, suggesting the presence of regulators of the enzyme activity. PMID- 9390184 TI - Partial purification and characterization of a Ca(2+)-dependent proteinase from Arabidopsis roots. AB - Ca2+, an important intracellular messenger in plants, is implicated in controlling diverse cellular functions by regulating the activity of several enzymes. Here we report the presence of a Ca(2+)-dependent proteinase (CDP) activity in roots of Arabidopsis using in-gel assays (zymograms). The CDP activity showed absolute Ca2+ requirement for its activation; other divalent ions such as Mg2+, Sr2+, and Zn2+ did not substitute for Ca2+ in stimulating protease activity. The CDP activity was inhibited by the proteinase inhibitors leupeptin, E-64, and N-ethylmaleimide, whereas pepstatin A and phenylmethylsulfonyl fluoride were without effect. These data indicate that the enzyme is likely to be a cysteine proteinase. The CDP activity was partially purified from root cultures using ammonium sulfate precipitation, DE-52, Mono-Q, and Superdex 200 column chromatography. This purification scheme resulted in about 40-fold purification of the CDP activity. Based on the elution of Arabidopsis CDP (ACDP) activity on gel filtration column the molecular mass of CDP was estimated to be about 75 kDa. Isoelectric focusing showed that the enzyme had a pI between 5.2 and 5.4. SDS polyacrylamide gel analysis showed that activity was associated with a 45-kDa polypeptide, suggesting that the native ACDP is a homodimer. Five different antibodies raised to animal CDPs did not cross-react with the partially purified protein. These data suggest that the plant CDP differs from the known CDPs characterized from animals and is likely to be a new CDP that is unique to plants. PMID- 9390185 TI - Structural changes of the sarcoplasmic reticulum Ca(II)-ATPase nucleotide binding domain by pH and La(III). AB - The Ca(2+)-ATPase from sarcoplasmic reticulum couples the hydrolysis of one molecule of ATP to the transport of two Ca2+ ions in skeletal muscle fibers. Here, we study the accessibility of the fluorescein covalently attached to the Lys515 at the nucleotide binding domain of the ATPase to the small collisional quencher iodide at pH 6 and 8, as well as the effect of ligand binding (La3+, La(3+)-nucleotide, and Ca2+). Our results indicate that bound fluorescein is significantly more accessible at pH 6 than at pH 8, suggesting that pH modulates the structure of the nucleotide binding domain of the ATPase. This notion was further substantiated by the finding that La(3+)-nucleotide only interacted with the catalytic center at acidic pH. Notably, the differential accessibility of the nucleotide binding domain at acidic and basic pH cannot be rationalized in terms of the ATPase E1/E2 conformational equilibrium since a shift of the ATPase toward the E1 (plus Ca2+) or E2 (plus EGTA) did not affect the accessibility of fluorescein-labeled ATPase to the quencher. Taken together, these findings show the presence of structural flexibility in the FITC binding site and suggest a structural modulation of the Ca(2+)-ATPase nucleotide binding domain by pH and La3+ binding through long-range link-age mechanisms. PMID- 9390186 TI - The role of cysteinyl residues in the activity of bacterial elongation factor Ts, a guanosine nucleotide dissociation protein. AB - The modification of E.coli elongation factor Ts (EF-Ts) by NEM and other sulfhydryl reagents inactivates the protein's ability to bind EF-Tu.GDP and to catalyze GDP exchange. The reactive residue was found to be Cys-22. Replacement of Cys-22 by Ser or Gly only partially impairs the binding or catalytic properties of EF-Ts while it completely protects EF-Ts from the inactivation by NEM. Cys-22 of EF-Ts is not located at the EF-Ts.EF-Tu interface, yet it can be modified only when EF-Ts is not bound to EF-Tu. These results support the proposal that the conformation change around Cys-22 in the amino terminus of EF Ts rather than Cys-22 itself is essential for binding EF-Tu. Apparently, modification of Cys-22 by NEM disrupts the conformation change and inactivates EF Ts. The return of EF-Ts to its native conformation may provide the driving force for the rate-determining step in the catalytic cycle, the dissociation of EF-Ts from EF-Tu.GNP. PMID- 9390187 TI - Hypoxia regulates xanthine dehydrogenase activity at pre- and posttranslational levels. AB - Hypoxia increases the activity of xanthine oxidase (XO) and its precursor, xanthine dehydrogenase (XDH), but the mechanism of regulation is unclear. In hypoxic Swiss 3T3 cells, an early (0-24 h) cycloheximide-insensitive increase in XO-XDH activity, coupled with a lack of increase in de novo XO-XDH synthesis (immunoprecipitation) or mRNA levels (quantitative RT-PCR), demonstrated a posttranslational effect of hypoxia. Similarly, hyperoxia decreased XO-XDH activity faster than could be accounted for by cessation of XO-XDH protein synthesis. In further support of a posttranslational effect, cells transfected with a constitutively driven XDH construct displayed an exaggerated increase in activity in hypoxia but no increase in activity in hyperoxia. However, more prolonged exposure to hypoxia (24-48 h) induced an increase in XO-XDH mRNA levels and de novo XO-XDH protein synthesis, suggesting an additional pretranslational effect. Finally, hypoxic induction of XO-XDH activity was found to be cell-type restricted. We conclude that control of XO-XDH levels by oxygen tension is a complex process which involves several points of regulation. PMID- 9390188 TI - Regulation of glucuronidation by glutathione redox state through the alteration of UDP-glucose supply originating from glycogen metabolism. AB - The effect of altered redox state of glutathione was investigated on p nitrophenol glucuronidation in isolated mouse hepatocytes. Decrease of GSH/GSSG ratio provoked by various agents caused increased glucuronidation which was accompanied by stimulated glycogenolysis and elevated UDP-glucose content. The stimulation of glycogenolysis and glucuronidation by glutathione consumption could be prevented by the reduction of oxidized glutathione with dithiothreitol and by the glycogenolysis inhibitor fructose. In permeabilized hepatocytes glycogen metabolism, bypassed by the addition of UDP-glucose, stimulated glucuronidation which was insensitive to glutathione depletion. In liver microsomes either UDP-glucuronosyltransferase activity or UDP-glucuronic acid transport was not influenced by GSH/GSSG ratio. These results suggest that alteration of the GSH/GSSG ratio regulates glucuronidation by affecting enzymes of the glycogen metabolism via the modification of UDP-glucuronate supply. PMID- 9390189 TI - Studies on the relationship between estrogen receptor content, glutathione S transferase pi expression, and induction by 2,3,7,8-tetrachlorodibenzo-p-dioxin and drug resistance in human breast cancer cells. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) induces both phase I and phase II drug metabolizing enzymes in rodent liver and hepatoma cell lines and this induction is mediated by the aryl hydrocarbon (Ah) receptor. Induction of CYP1A1 by TCDD in human breast cancer cells has been reported and results of several studies suggest that the estrogen receptor (ER) may be required for Ah responsiveness. This study investigates the induction of GST pi by TCDD in human breast cancer cells and the role of the ER in mediating this response. TCDD did not induce chloramphenicol acetyl transferase (CAT) activity in ER positive (ER+) MCF-7 and ER- MDA-MB-468 and MDA-MB-231 human breast cancer cell lines transiently transfected with GST pi (human) or GSTP (rat) promoter-reporter constructs containing the -291/+36 and -2.9/+59 region, respectively, of the GST pi and GSTP gene promoters. Furthermore, TCDD did not induce GST pi or GSTP in MDA-MB-468 and MDA-MB-231 human breast cancer cells stably transfected with the ER. RT-PCR confirmed that GST pi mRNA levels were low in ER+ MCF-7 cells and high in ER- MDA MB-468 and MDA-MB-231 cells; however, in MDA-MB-468 and MDA-MB-231 cells stably transfected with the ER GST pi mRNA levels remained elevated and were not inducible. MDA-MB-468 and MDA-MB-231 cells stably transfected with the ER exhibited increased GST activity and decreased GSH content compared to wild-type cells; however, in MDA-MB-468 cells stably transfected with ER, the susceptibility to doxorubicin, ellipticine, chlorambucil, malphalan, or cisplatin was similar to that observed in wild-type cells. Adriamycin accumulation was similar in wild-type and ER stably transfected cells and verapamil did not affect this response, suggesting that ER expression did not influence p-glycoprotein activity. Taken together these data suggest that not all GST isoforms are responsive to TCDD and stable transfection of ER- cells with ER is not sufficient to restore the ER+ phenotype in some breast cancer cell lines. PMID- 9390190 TI - Subunit interactions of Escherichia coli F1-ATPase: mutants of the gamma subunits defective in interaction with the epsilon subunit isolated by the yeast two hybrid system. AB - Previously, we established a method to detect subunit interactions of F1-ATPase by the yeast two-hybrid system (Moritani, C., et al. Biochim. Biophys. Acta 1274, 67-72, 1996). Here, we isolated mutants of the gamma subunits defective in interaction with the epsilon subunit by this new procedure to study the molecular basis of coupling mechanisms of the F1F0-ATPase. Based on the intensities of the reporter gene expression in this system, five mutants of the gamma subunit with different levels of gamma-epsilon interactions were isolated and their single base substitutions were determined. Mutants with a substitution of Pro-55 for Leu, Thr-102 for Met, Val-141 for Asp, or Gln-235 for Leu exhibited decreased reporter gene expression, suggesting decreased levels of interaction, while Asp 85 for Gly mutation caused a higher level of expression, suggesting increased interaction. Among these point mutations, G85D, M102T, or D141V mutations were introduced into the gamma subunit gene in the plasmid carrying whole unc operon. Transformants carrying a deletion mutant of the whole unc operon with these expression plasmids were analyzed. Mutations M102T and D141V with decreased gamma epsilon interaction caused increases of membrane-bound F1-ATPase activity and proton pumping activity, while G85D with increased gamma-epsilon interaction exhibited lower levels of F1-ATPase activity in the membranes. Molecular assembly of the F1 subunits on the mutant membranes detected by Western blotting exhibited no defect for all three mutants. These results suggested that the correlation between the ATPase activity and gamma-epsilon interaction is reciprocal and this interaction may regulate the ATPase activity. The topological and functional importance of Gly-85, Met-102, and Asp-141 together with Leu-55 and Leu-235 in gamma-epsilon interaction is discussed. PMID- 9390191 TI - The Ah receptor is a sensitive target of geldanamycin-induced protein turnover. AB - Geldanamycin (GA) binds directly to hsp90 and apparently disrupts certain hsp90 heterocomplexes. We have investigated the GA-hsp90 interaction and its effect on other associated proteins. Incubation of 2-[125I]-iodo-3-azido-7,8-dibromo-p dioxin-labeled Hepa 1c1c7 cytosol with GA-coupled beads revealed a stable association of Ah receptor (AhR)/hsp90 complex with GA. In addition, sucrose gradient sedimentation analysis demonstrated that GA does not disrupt the 9S Ah receptor complex in vitro. HeLa and Hepa 1c1c7 cells were subjected to a dose response and time-course treatment with GA and the level of the AhR was determined. A 75% depletion in AhR levels was observed within an hour of exposure to 100 nM GA. The relative stability of other proteins that associate with hsp90 was determined with the following rank order of sensitivity to GA exposure: AhR >> c-Raf-1 > glucocorticoid receptor > CDK4 >> p50. A series of hsp90 deletion mutants were used to map the domain that interacts with GA. Deletion of the first 221 amino acids in NH2-terminal domain resulted in loss of binding to solid-phase GA. Epitopes of monoclonal antibodies specific for hsp90 were also determined by direct immunoprecipitation with hsp90 mutants. Results indicated that monoclonal antibodies 8D3 and 3G3 interact with hsp90 via the first 221 amino acids in NH2 terminal region, whereas AC88 requires a COOH-terminal region between amino acids 661-677. PMID- 9390192 TI - Maintenance of transfection rates and physical characterization of lipid/DNA complexes after freeze-drying and rehydration. AB - It is well established that cationic liposomes form complexes with DNA and effectively transfect cells in vivo and ex vivo. Lipid/DNA complexes have proven safe and nonimmunogenic in clinical trials; however, they are known to aggregate readily in liquid formulations. This physical instability requires clinicians to prepare lipid/DNA complexes immediately prior to injection. In order to eliminate problems associated with this temporal requirement, we investigated the feasibility of preserving complexes as a dried preparation that could be tested, stored, and rehydrated as needed. To this end, our study evaluated the ability of different stabilizers to preserve transfection rates of complexes during acute freeze-drying stress. Our data show that complexes lyophilized in 0.5 M sucrose or trehalose possessed transfection rates similar to those of fresh preparations. In addition, dried complexes that exhibited full transfection activity upon rehydration had sizes comparable to nonlyophilized controls. Our work demonstrates that lipid/DNA complexes can be stabilized as dried powders that offer significant advantages over current liquid formulations. Furthermore, the correlation of transfection rates with maintenance of complex diameter suggests that size plays a critical role in lipid-based DNA delivery. PMID- 9390193 TI - Identification of the subunits and target peptides of pig heart NAD-specific isocitrate dehydrogenase modified by the affinity label 8-(4-bromo-2,3 dioxobutylthio)NAD. AB - Pig heart NAD-dependent isocitrate dehydrogenase reacts with 8-(4-bromo-2,3 dioxobutylthio)-NAD (8-BDB-TNAD) with incorporation of 1.21 mol of reagent/mol of average subunit when the enzyme reaches the limit of 25% residual activity (Kumar, A., and Colman, R. F., Arch. Biochem. Biophys. 308, 357-366, 1994). Inclusion of NADPH decreases both the extent of inactivation and the reagent incorporation to 0.55 mol/mol of average subunit. We have now isolated the peptides labeled by radioactive 8-(4-bromo-2,3-dioxobutylthio)-[2-3H]NAD and have located them within the sequence of pig heart NAD-dependent isocitrate dehydrogenase. The enzyme is composed of three types of subunits, present as alpha 2 beta gamma. We have separated the subunits from unmodified and 8-BDBT[2 3H]NAD-modified enzymes by HPLC on a C4 reverse-phase column, after pretreatment of the enzymes with sodium dodecyl sulfate or urea, and compared the subunit sequences of the porcine enzyme with those of the corresponding subunits from other mammalian NAD-dependent isocitrate dehydrogenases. The predominant radioactivity of 8-BDBT[2-3H]NAD is observed in the alpha and gamma peaks, and the NADPH-protected enzyme exhibits marked reduction in incorporation into these peaks. However, evidence based on recombination of subunits from modified and unmodified enzymes indicates that only labeling of the alpha-subunit is responsible for inactivation by 8-BDB-TNAD. Cyanogen bromide was used to cleave the modified enzyme, and we purified one labeled peptide from the alpha-subunit (amino acids 84-177) as well as one from the gamma-subunit (amino acids 67-186). In the alpha-subunit, decreased modification by [7-14C]-phenylglyoxal of Arg88 and Arg98 after prior labeling of the enzyme by 8-BDB-TNAD indicates that these residues are the critical target sites of the reactive nucleotide analogue. We conclude that alpha subunit's Arg88 and Arg98 are both at or near the allosteric NADPH sites of the pig heart isocitrate dehydrogenase. PMID- 9390194 TI - Substrate specificity and characterization of partially purified rat liver 13 hydroxyoctadecadienoic acid (13-HODE) dehydrogenase. AB - Oxidation products of linoleic acid, such as 13-hydroxyoctadecadienoic acid (13 HODE), exhibit biological activity in a number of systems. One major metabolic fate of 13-HODE is oxidation to the 2,4-dienone, 13-oxooctadecadienoic acid by an NAD(+)-dependent dehydrogenase (13-HODE dehydrogenase). The present work describes the partial purification and characterization of 13-HODE dehydrogenase from rat liver cytosol. The enzyme was purified using a combination of ammonium sulfate precipitation, as well as hydroxylapatite, gel permeation, and hydrophobic interaction chromatography. Analysis of the most purified preparation by SDS-polyacrylamide gel electrophoresis indicates two subunits of approximately 55 kDa, suggesting the possibility of a heterodimeric enzyme. However, due to aggregation in the purified preparation, an accurate molecular mass for the native enzyme has not yet been obtained. Using 13-HODE as a substrate, the purified enzyme has a Km of 6.3 microM and a Vmax of 5.7 nmol/min/mg. More importantly, the enzyme has a narrow substrate specificity with 13-HODE being the preferred substrate. From a series of 17 potential substrates, only 9-HODE (53% the activity of 13-HODE) and 15-hydroxyeicosatetraenoic acid (64% the activity of 13-HODE) showed significant activity as substrates. A number of other unsaturated hydroxy fatty acids, including several eicosanoids, are not substrates. The narrow substrate specificity displayed by the enzyme suggests that it could play a key role in modulating the effects of oxidized derivatives of linoleic acid. PMID- 9390195 TI - The homeodomain protein Pbx1 is involved in cAMP-dependent transcription of human CYP17. AB - Pbx1 is a homeodomain transcription factor involved in cAMP-dependent transcriptional regulation of the bovine CYP17 gene. In this study, we have investigated the involvement of Pbx1 in the transcriptional regulation of the human CYP17 gene. Although a sequence identical to previously determined Pbx binding sites is not present in the promoter region of the human CYP17 gene, three putative Pbx-binding sites are identified by sequence similarity analysis. Coexpression of Pbx1 and a catalytic subunit of protein kinase A (PKA) greatly enhances reporter gene transcription via the 5'-flanking region of the human CYP17 gene. Upon gel shift analysis utilizing nuclear extracts from human adrenal H295R cells, one of the three putative Pbx1-binding sites, -250/-241 bp, shows the typical intense doublet observed with other Pbx-binding sites. 5'-Deletion analyses of the reporter construct containing this Pbx-binding site showed approximately sixfold induction by coexpression of Pbx1 and PKA compared to the basal transcription, suggesting that Pbx1 binds the -250/-241 bp sequence and participates in cAMP-dependent regulation of the human CYP17 gene. PMID- 9390196 TI - The tumor-bearing state induces augmented responses of organ-associated lymphocytes to high-dose interleukin-2 therapy in mice. AB - A high-dose bolus regimen for interleukin(IL)-2 administration to cancer patients frequently causes serious side-effects in which various organs are involved. In order to reveal the mechanism of toxicities associated with this regimen, we compared the augmenting effect of high-dose IL-2 on murine organ-associated lymphocytes between neoplastic and non-neoplastic states. Intraperitoneal administration of IL-2 at a dose of 10(5) JRU (Japanese Reference Units) twice daily for 3 days led to the death of all the syngeneic MH134-hepatoma- or X5563 myeloma-bearing mice, whereas it had no lethal effect on non-tumor-bearing mice. Histological and morphometric analyses demonstrated that tumor-bearing mice displayed more extensive infiltration of large granular lymphocytes and agranular lymphocytes in the liver and lungs than did the non-tumor-bearing mice. Large granular lymphocytes had the ultrastructural characteristics of lymphokine activated killer cells. Lymphocytes often underwent extravasation into the interstitial space and exhibited local proliferation without causing any direct injury to apposed parenchymal cells. Flow-cytometric analysis of hepatic mononuclear cells demonstrated that IL-2-receptor-beta (IL-2R beta)-bearing lymphocytes, i.e., natural killer cells and intermediate CD3 cells, were increased in number in the neoplastic state before the IL-2 injection. The present study indicates that the tumor-bearing state increases the number of organ-associated IL-2R beta + lymphocytes, which are then greatly amplified by the challenge of high-dose IL-2, leading to the functional disturbance of organs. We have further demonstrated here that an intermittent low-dose IL-2 regimen has a potential therapeutic effect on tumor regression without causing lethal side effects. PMID- 9390197 TI - Restoration of macrophage tumoricidal activity by bleomycin correlates with the decreased production of transforming growth factor beta in rats bearing KDH-8 hepatoma cells. AB - To explore the mechanisms of immuno-modulatory activities of bleomycin, we investigated interferon gamma (IFN gamma) mRNA expression, tumor necrosis factor alpha (TNF alpha) production, nitric oxide (NO) production and macrophage tumoricidal activities in rats bearing KDH-8 hepatoma cells, which secreted a large amount of transforming growth factor beta (TGF beta), and these processes in KDH-8 tumor-bearing rats treated with bleomycin. We found that IFN gamma mRNA expression, TNF alpha production, NO production and macrophage cytotoxic activities were lower in the KDH-8-bearing rats than in normal rats. On the other hand, low-dose bleomycin restored the macrophage cytotoxic activities, NO production, IFN gamma mRNA expression and TNF alpha production in the KDH-8 bearing rats. In vitro experiments showed that KDH-8-derived TGF beta decreased the IFN gamma mRNA expression and TNF alpha production in splenocytes, and NO production in peritoneal macrophages. These results suggest that low-dose bleomycin restored the cytokine production and macrophage tumoricidal activities in the KDH-8-bearing rats by decreasing KDH-8-derived TGF beta. PMID- 9390198 TI - Treatment of recurrent glioma with intracavitary alloreactive cytotoxic T lymphocytes and interleukin-2. AB - For a single-dose toxicity assessment, five patients with recurrent malignant glioma (ages 29-46 years) were treated with intracavitary alloreactive cytotoxic T lymphocytes (CTL) and interleukin-2 (IL-2). The trial tested the hypothesis that alloreactive CTL, sensitized to the major histocompatibility complex (MHC) proteins of the patient, offer selective, targeted killing of glioma cells that express MHC. Patient lymphocytes, which also express MHC, were irradiated and placed into CellMax artificial capillary systems with lymphocytes from MHC disparate donors and CTL developed over a 2- to 3-week period with a low concentration of IL-2. The CTL largely expressed CD3 and CD11a/CD8 markers and lysed targets displaying patient MHC. CTL were implanted into the tumor bed at surgery and a catheter was used for subsequent infusions. Patients received one to five treatment cycles every other month; one cycle generally consisted of two or three CTL infusates administered within a 1- to 2-week period. Different unrelated donors were used for each cycle. Treatment was well tolerated; transient toxicity at grades 1-3 was recorded by NCI Common Toxicity Scale criteria. Two glioblastoma patients have died; one from tumor recurrence locally and the other from recurrence at a site distant from the treatment. Two of the five patients completed five cycles; one anaplastic oligodendroglioma patient shows no evidence of tumor 30 months from the start of immune therapy and an anaplastic astrocytoma patient shows stable disease 28 months after initiation of therapy. One anaplastic oligodendroglioma patient, who dropped the protocol during her second treatment cycle, has no evidence of tumor 28 months after recurrence. PMID- 9390199 TI - Sequence variation in the monoclonal-antibody-U36-defined CD44v6 epitope. AB - Monoclonal antibody (mAb) U36 was developed for the treatment of minimal residual disease of head and neck squamous cell carcinoma (HNSCC). The mAb-U36-defined antigen was characterized by cDNA cloning, and was shown to be identical to the keratinocyte-specific CD44 splice variant epican. The epitope recognized by mAb U36 was shown to be located in the v6 domain. Two amino acids within the epitope appeared to differ from the sequences that have been described in literature. The sequence of the epitope appeared to contain glutamic acid at position 367 and lysine at position 374, while valine and arginine respectively have been described before. Interestingly, another anti-CD44v6 antibody with possible clinical application, VFF18, recognizes an epitope in the same area. With respect to the applicability of these antibodies for tumor targeting, this variation might have an influence on antibody-antigen interaction and mAb accumulation in the tumor. Furthermore, this observation raised the question whether the different epitopes are related to the malignant behavior of tumor cells. In this paper we determine the relative affinity of mAb U36 for the variant epitope sequences by tumor cell binding assays using synthetic peptides for competition. The presence of glutamic acid instead of valine at position 367 caused strong competition. Further evaluation showed that the published valine variant does not exist in vivo, and is the result of a sequencing artefact. The effect of substitution of lysine for arginine at position 374 had no effect on the binding of mAb U36 to the cells. This amino acid variation was shown to be due to allelic polymorphism. There was no trend towards allelic imbalance in tumor cells as compared to normal cells. PMID- 9390200 TI - Intrapleural instillation of interferon gamma in patients with malignant pleurisy due to lung cancer. AB - The effect of intrapleural instillation of recombinant human interferon gamma (IFN gamma) at increasing doses of (1-12) x 10(6) U was examined in six patients with cytologically positive pleural effusion due to lung cancer. Intrapleural instillation was repeated up to three times. Clinically, no reaccumulation of pleural effusion was observed in one patient and disappearance of lung cancer cells from the pleural effusion was seen in two other patients. No severe side effects were observed. Considerable levels of IFN gamma remained in the pleural effusion as well as in patients' serum up to 7 days after instillation of 2 x 10(6) U and higher doses. The total cell number showed a transient decrease on day 1 of therapy. Levels of pro-inflammatory cytokines, such as tumor necrosis factor alpha, interleukin(IL)-1 beta and IL-6, in the pleural effusion remained almost stable after IFN gamma instillation. On the other hand, intrapleural IL-1 receptor antagonist levels were remarkably elevated by the instillation of IFN gamma. IL-2- and IL-12-inducible killer activity of pleural mononuclear cells tended to increase slightly. Despite the inability of IFN gamma to control pleural effusion in this treatment schedule, IFN gamma instilled by an intrapleural route had a potential local antitumor activity. Moreover, since IFN gamma persists in pleural effusions for a long time after a single instillation, such a therapy in combination with other fibrogenic biological response modifiers can be promising. PMID- 9390201 TI - Antitumor effects of the combination immunotherapy with interleukin-12 and tumor necrosis factor alpha in mice. AB - There is strong evidence that antitumor activity of interleukin-12 (IL-12) in vivo is mediated, in part, through interferon (IFN gamma) produced by IL-12 stimulated natural killer and T cells. Since IFN gamma and tumor necrosis factor alpha (TNF alpha) have been reported to synergize in antitumor effects in a number of models, we decided to examine whether the combined treatment with recombinant mouse IL-12 and recombinant human TNF alpha would produce similar effects. The efficacy of the combined IL-12/TNF alpha immunotherapy was evaluated in three tumor models in mice: B16F10 melanoma, Lewis lung (LL/2) carcinoma and L1 sarcoma. Intratumoral daily injections of 1 microgram IL-12 in combination with 5 micrograms TNF alpha into B16F10-melanoma-bearing mice resulted in a significant retardation of the tumor growth as compared with that in controls and in mice treated with either cytokine alone. Similar effects were obtained using 0.1 microgram IL-12 and 5 micrograms TNF alpha in LL/2 carcinoma and L1 sarcoma models. Antitumor activity against L1 sarcoma was still preserved when TNF alpha at a low dose (1 microgram) was combined with 0.1 microgram IL-12 and applied for a prolonged time. Potentiation of antitumor effects, which was observed in IL 12/TNF alpha-based immunotherapy, could result from at least three different mechanisms, partly related to stimulation of IFN gamma and TNF alpha production in treated mice: (a) direct cytostatic/cytotoxic effects on tumor cells, (b) induction of antitumor activity of macrophages, and (c) inhibition of blood vessel formation in the tumor. Our studies demonstrate that combination tumor immunotherapy with IL-12 and TNF alpha may be more effective than single-cytokine treatment, and suggest possible mechanisms by which IL-12 and TNF alpha may exert potentiated therapeutic effects against locally growing tumors. PMID- 9390202 TI - Active and indolent chronic lymphocytic leukaemia--immune and hormonal peculiarities. AB - A group of 138 B cell chronic lymphocytic leukaemia (B-CLL) patients, 83 with active disease and 53 having the indolent form of the disease, were evaluated. The aim of the study was to clarify whether indolent and active B-CLL differ in their immune and hormonal characteristics. Peripheral blood lymphocyte proliferation in response to phytohaemagglutinin, concanavalin A, recombinant interleukin-2, dextran sulphate, Pisum sativatum agglutinin and wheat germ agglutinin was investigated. Serum immunoglobulin and beta 2 microglobulin levels were determined. Adrenocorticotropic hormone (ACTH), cortisol, follicle stimulating hormone luteinizing hormone, 17 beta-oestradiol, testosterone, triiodothyronine, thyroxine, thyroglobulin and thyrotropic hormone levels were determined by radioimmunoassay. Active and indolent CLL presented differences in immunological characteristics, as demonstrated by the more severe suppression of T lymphocyte function, reduced IgA level and considerably higher serum beta 2 microglobulin values in active disease. Immune disturbances were accompanied by hormonal imbalance, depending on disease status: lower ACTH, cortisol and triiodothyronine levels were established to occur in active CLL compared to indolent disease. Male patients demonstrated striking changes in sex hormones, which were more evident in active disease. The findings point to the complexity of immuno-hormonal disturbances in CLL with differences in the active and indolent state of the disease. PMID- 9390203 TI - The antitumor effects of levamisole in mice are mediated by NC-1.1+ cells. AB - Murine natural cytotoxicity, which is a major component of the innate immune response in cancer, is mediated by leukocytes that express the NC-1.1 receptor. Mice depleted of natural cytotoxicity by treatment with an anti-NC-1.1 mAb show enhanced growth of certain transplantable tumors, so agents that enhance natural cytotoxicity by NC-1.1+ cells have the potential to be effective anticancer therapeutic agents. We have examined the immunomodulatory effect of levamisole on natural cytotoxicity mediated by NC-1.1+ cells against the BALB/c WEHI-164 murine fibrosarcoma. Administration of levamisole to BALB/c mice significantly enhanced in vitro splenic natural cytotoxicity against 51Cr-labeled WEHI-164 tumor cells. The effect was most marked 48 h after levamisole treatment, at a dose of 10 mg/kg body weight. This enhancement of natural cytotoxicity by levamisole could be completely abrogated by pretreatment of mice with an anti-NC-1.1 mAb. Treatment of BALB/c mice with 10 mg/kg levamisole significantly reduced the growth of WEHI 164 and this effect was abrogated by pretreatment of mice with anti-NC-1.1, indicating that the antitumor effect of levamisole was mediated, at least in part, via NC-1.1+ cells. PMID- 9390204 TI - Utility of hyaluronic acid in pleural fluid for differential diagnosis of pleural effusions: likelihood ratios for malignant mesothelioma. AB - The level of hyaluronic acid (HA) was determined in the pleural fluid of 99 patients, including 19 with malignant mesothelioma, 27 with lung cancer, 1 with breast cancer, 1 with mediastinal tumor and 51 with non-malignant diseases. With a cut-off level at 100 micrograms/ml, the pleural fluid concentration of HA was high in 36.8% of patients (7 of 19) with malignant mesothelioma and 1.3% of patients (1 of 80) with lung cancer and other malignant and non-malignant diseases. The mean concentration of pleural fluid HA was significantly higher in patients with mesothelioma than in those with lung cancer and other malignant and non-malignant diseases. The pre-test probability of MM was 5.9% in this series. The LRs for > or = 100, 50-99 and < or = 49 micrograms/ml are 28.3, 3.3 and 0.5, respectively; these put the post-test probabilities at 64, 17 and 3%, respectively. Indeed, in cases of uncommon disease such as MM, the post-test probability is low even if the cut-off level of HA is > or = 100 micrograms/ml. The discrimination between malignant mesothelioma and lung cancer needs special attention. In these two diseases, the LRs of MM for pleural fluid CEA > 30, 10-30 and < 10 ng/ml were 0.2, 1.9 and 2.4, respectively. The pre-test probability of MM for HA > or = or 100 micrograms/ml is 64%. Furthermore, because the LR for CEA is < 10 ng/ml, the post-test probability is 81%. When the combination of two markers is considered, the high level of HA and the low level of CEA may be useful for the differential diagnosis of MM from pleuritis carcinomatosa. PMID- 9390205 TI - Prognostic significance of argyrophilic nucleolar organizer region (AgNOR) in resected non-small cell lung cancer (NSCLC). AB - The expression of the nucleolar organizer regions (NORs) was quantified in paraffin sections of tumors and lymph node metastasis, by means of digital image analysis, in 75 patients with resected non-small cell lung cancer (NSCLC). Patients were divided in two groups: early stage (stages I and II) and advanced stage (stages IIIa, IIIb and IV). The prognostic significance of AgNOR expression was tested by Cox regression analysis in models controlled for age, sex, vital status, stage and histological type. Tumors at early stages had a lower expression of AgNOR than those at more advanced diseases. The mean values obtained for NORs in advanced disease were almost the same as those in the primary tumors when compared with the corresponding lymph node metastasis (r = 0.90; p < 0.01; linear regression). The prognostic role of AgNOR was significant only for tumors at stages I and II and not for advanced neoplasms (stages IIIa, IIIb and IV). These results encourage the inclusion of AgNOR quantitation in routine material, especially in early lung cancer. PMID- 9390206 TI - Revisit of primary malignant neoplasms of the trachea: clinical characteristics and survival analysis. AB - A clinical review with an analysis of prognostic factors, including clinical characteristics, histological classifications, presenting symptoms/signs and treatment modalities, was conducted in 67 patients with primary malignant neoplasms of the trachea who were seen at the Veterans General Hospital-Taipei between 1979 and 1994. The incidence of tracheal cancer was 140 times less than lung cancer during this period. Delayed diagnosis of more than 6 months after onset of symptoms occurred in one-third of the patients owing to lack of specific symptoms/signs indicative of tracheal disease. Epidermoid carcinoma was the most frequent histological type encountered and accounted for half of the cases. Surgical resection was the first choice of treatment for all patients if the disease was locally confined, except for small cell carcinoma and lymphoma. Radiotherapy was given if the patient was not suitable for surgery. Single and multivariate analyses showed that clinical symptoms and signs were not related to prognosis, except for general malaise and acute respiratory failure. Patients suffering from adenoid cystic carcinoma and mucoepidermoid carcinoma had a better prognosis than other histological diagnoses. Whether the patient received radiotherapy or not proved to be a significant prognostic factor in the patients. Patients with tracheal cancers had a poorer prognosis than those with lung cancer. PMID- 9390207 TI - Feasibility of adjuvant chemotherapy for breast cancer patients. AB - To evaluate the feasibility of adjuvant chemotherapy, we analyzed the toxicities of chemotherapy for primary breast cancer in Japanese women. Since the opening of the National Cancer Center Hospital East, 180 female breast cancer patients have received adjuvant chemotherapy or chemo-hormonal therapy following surgical treatment between June 1992 and December 1995. On the basis of informed consent about prognosis and adjuvant therapy, most patients decided to choose the type of cytotoxic chemotherapy themselves. Adjuvant chemotherapy consisted of oral fluoropyrimidine compounds (OFP), cyclophosphamide + adriamycin +/- 5 fluorouracil [CA(F)] or cyclophosphamide + methotrexate + 5-fluorouracil (CMF). Toxicity was determined using the Toxicity Grading Criteria of the Japan Clinical Oncology Group (JCOG). Sixty-six patients received OFP, 59 CA(F) and the rest 55 CMF. The toxicity grading of leukocytes and neutrophils was significantly higher in patients treated with CA(F) or CMF than in those treated with OFP. Similar results were also seen relating to the toxicity of nausea/vomiting and alopecia. There was no statistical difference in the toxicity grading of hemoglobin, glutamic oxaloacetic transaminase/glutamic pyruvic transaminase (GOT/GPT) and stomatitis/ gastritis between the three groups of patients. Interestingly, the number of patients that were forced to discontinue chemotherapy was higher in those receiving OFP than in those receiving CA(F) or CMF. Cytotoxic chemotherapy of CA(F) or CMF results in greater toxicity than OFP, but is tolerated and feasible in the adjuvant setting used in Japanese breast cancer patients from the viewpoint of toxicities by anticancer chemotherapy. PMID- 9390208 TI - Treatment of previously treated metastatic breast cancer by mitoxantrone and 48 hour continuous infusion of high-dose 5-FU and leucovorin (MFL): low palliative benefit and high treatment-related toxicity. AB - For previously treated advanced breast cancer, there is no standard second-line therapy. Combination chemotherapy with mitoxantrone, high-dose 5-fluorouracil (5 FU) and leucovorin (MFL regimen) had been reported as an effective and well tolerated regimen. From October 1993 to November 1995, we treated 13 patients with previously chemotherapy-treated metastatic breast cancer by mitoxantrone, 12 mg/m2, on day 1 and continuous infusion of 5-FU, 3000 mg/m2, together with leucovorin, 300 mg/m2, for 48 h from day 1 to 2. Each course of chemotherapy was given every 4 weeks. Most of these patients had more than two metastatic sites, with lung metastasis predominant. Seven patients had been treated with anthracycline. Seven patients had previously received radiotherapy and seven had received hormone therapy. Median number of courses of MFL regimen given was six and the median cumulative dose of mitoxantrone was 68.35 mg/m2. One patient had complete response, seven had stable disease, none had partial response and five had progressive disease. The overall objective response rate was 7.6%. The median follow-up period was 14 months. Median survival was 16 months. Median progression free survival was 5 months. A complete responder had relapse-free survival up to 17 months. Major toxicities were cardiotoxicity and leukopenia. Eight patients were dead in the last follow-up; two of them died of treatment-related toxicity. The MFL regimen achieves little palliative benefit and induces severe toxicity at a fairly high rate. Administration of this regimen to breast cancer patients who have been treated by chemotherapy and those with impaired heart function requires careful attention. PMID- 9390209 TI - Predictive factors for tumor response to systemic chemotherapy in patients with hepatocellular carcinoma. AB - Chemotherapy is of limited value in the treatment of hepatocellular carcinoma (HCC), since there are no established chemotherapeutic regimens proven to be effective. The aim of the present study was to determine predictive factors for tumor response to systemic chemotherapy in HCC patients. The relationship between patients' characteristics and tumor response was examined in 147 previously untreated HCC patients receiving systemic chemotherapy. Ten patients showed partial response (PR) and none showed complete response (CR). The response rate for all single anticancer agents was less than 10% and the overall response rate was 6.8%. The response rate in patients with unilateral HCC was significantly higher than in those with bilateral HCC. However, there were no responders among patients with a performance status of 2-3, ascites, a tumor occupying more than 50% of the entire liver, tumor thrombus in the main portal trunk or a serum bilirubin level of more than 2.0 mg/dl. There was a close relationship between patients' characteristics and tumor response. It is concluded that patients with fairly advanced HCC and/or poor hepatic reserve should not be given systemic chemotherapy. PMID- 9390210 TI - A phase II study of high-dose epirubicin (EPI) plus cyclophosphamide (CPA) with G CSF for breast cancer patients with visceral metastases or hormone-independent tumors: a trial of the Japan Clinical Oncology Group. AB - To evaluate the efficacy and toxicity of high-dose epirubicin (EPI) plus cyclophosphamide (CPA) therapy, a phase II study of EPI, 130 mg/m2, plus CPA, 1000 mg/m2, with G-CSF every 3 weeks was carried out for 51 advanced or recurrent breast cancer patients by the Japan Clinical Oncology Group (JCOG). Fifty out of the 51 patients who were eligible for our criteria were treated with this regimen as first-line chemotherapy for visceral metastases or hormone-independent tumors. In this trial, 203 cycles were administered with an average of four cycles per patients. In 50 patients who were evaluable for response, there were 7 complete (CR) and 25 partial responses (PR) with an overall response rate of 64% (95% confidence interval, 50.1-75.9%). Symptomatic and hematological acute toxicity more than grade 3 occurred frequently; however, no treatment-related death occurred. The incidence of toxicities (> or = grade 3) was as follows: leukopenia 98%, thrombocytopenia 42%, nausea/vomiting 56% and hair loss 12%. In each cycle, daily administration of 2 micrograms/kg G-CSF (granulocyte-colony stimulating factor) was given on days 2-15 subcutaneously. The incidence of cardiotoxicity was low. Arrhythmia (< or = grade 2) was observed in 8% and a slight decrease of ejection fraction index (< or = grade 2) was observed in 2% in this trial. The median follow-up period for patients was 37.2 (24.6-51.5) months and the median survival period was 17.4 months. These data indicate that high-dose EPI + CPA combination chemotherapy was effective and well tolerated for breast cancer patients with visceral metastases or hormone-independent tumors. A randomized trial of high-dose EPI vs conventional chemotherapy is required to ascertain the usefulness of this regimen. PMID- 9390211 TI - A pilot study of multimodality therapy for initially unresectable liver metastases from colorectal carcinoma: hepatic resection after hepatic arterial infusion chemotherapy and portal embolization. AB - The prognosis of patients with unresectable liver metastases is poor, even if hepatic arterial infusion chemotherapy (HAI) or systemic chemotherapy is administered. A pilot study was performed to evaluate the feasibility and efficacy of multimodality therapy with hepatectomy after HAI and portal embolization for such patients. Eight patients with colorectal carcinoma and synchronous unresectable liver metastases underwent resection of the primary tumor and placement of a pump, followed by HAI with 5-fluorouracil and mitomycin C. Owing to shrinkage of the liver metastases, two patients could undergo extended right hepatic lobectomy after portal embolization, which was deemed to be essential to prevent post-operative hepatic failure. The median survival time of the eight patients was 30 months, with a response rate of 75%. Complications including sclerosing cholangitis and duodenal ulcer were observed in five patients (63%). Additional hepatectomy could be performed successfully after portal embolization without morbidity in two patients. These two patients are still alive more than 6 years after initiation of HAI and have been free of disease for more than 5 years after hepatectomy. Hepatectomy after HAI and portal embolization is feasible and may be an option to cure selected patients with initially unresectable liver metastases. PMID- 9390212 TI - Prolonged survival in a nasopharyngeal carcinoma patient with multiple metastases: a case report and review of the literature. AB - Nasopharyngeal carcinoma is a common cancer in South East Asia. In the early stages, radiotherapy alone may achieve sustained control, but once metastasis occurs, it becomes an incurable disease with limited survival time. We report a case of nasopharyngeal carcinoma, initial stage T4N0M0, diagnosed in 1985 in a patient aged 36 years who received 70 Gy radiotherapy to the head and neck region. In 1988, relapse occurred with multiple lung metastases. The patient received many chemotherapy regimens with a very good response, including near complete remission with the first treatment regimen of cisplatin, 5-fluorouracil and leucovorin for lung metastases, and with the fifth chemotherapy regimen of ifosfamide as a single agent. After ifosfamide treatment, there was residual fibrotic change in the lung and complete disappearance, lasting for almost a year, of the liver and bone lesions. The patient eventually died in July 1995 due to progressive disease. Prolonged survival after mainly thoracic metastasis is possible in patients with nasopharyngeal carcinoma, especially if the tumor is chemo-responsive. PMID- 9390213 TI - Thyroglossal duct carcinoma: a case report. AB - We describe a 47-year-old woman with a 13-year history of asymptomatic midline submental swelling. Cytologic examination of a fine-needle aspiration specimen from the solid mass revealed adenocarcinoma. The preoperative diagnosis was thyroglossal duct carcinoma. A Sistrunk procedure was performed and microscopic examination revealed papillary adenocarcinoma. The postoperative course was uneventful and there were no signs of local recurrence or metastasis at one year after surgery. PMID- 9390214 TI - Hepatocellular carcinoma with gastrointestinal hemorrhage caused by direct tumor invasion to the duodenum. AB - Gastrointestinal hemorrhage from hepatocellular carcinoma invading the duodenum is very rare. A 60-year-old man with multiple hepatocellular carcinoma was admitted to our hospital because of massive melena and hematemesis. We succeeded in hemostasis of an esophageal variceal rupture by endoscopic varicial ligation. The duodenum could not be observed endoscopically due to extramural compression to the stomach from the liver tumor. Massive gastrointestinal hemorrhage occurred again and the patient died of hepatic failure. The postmortem examination revealed that the liver tumor had invaded the second portion of the duodenum and perforated into the lumen. PMID- 9390215 TI - Pheochromocytoma growing exophytically from the right adrenal gland and invaginating into the liver. AB - A 5-cm pheochromocytoma located in segment 7 of the liver was found incidentally in a 45-year-old man with mild diabetes mellitus and hypertension, and resected. The tumor was demonstrated by computed tomography and magnetic resonance imaging to have completely invaginated itself into the liver and to be receiving blood from a dilated right hepatic artery alone. Surgery revealed the hepatic mass to be tightly adherent to the right adrenal gland. The histopathologic diagnosis was pheochromocytoma growing exophytically from the right adrenal gland. There was no association with multiple endocrine neoplasia type 1 and type 2. A postoperative 131I metaiodobenzylguanidine scan revealed no accumulation, and the patient is currently doing well without recurrence or hypertension one year after the operation. A pheochromocytoma deeply invaginating into the liver should be considered in the differential diagnosis of primary hypervascular hepatic tumors. PMID- 9390216 TI - Adenomyomatous polyp of the uterus in a patient receiving tamoxifen. AB - We report a case of adenomyomatous polyp that developed during treatment with tamoxifen for breast cancer. A 63-year-old Japanese woman was admitted complaining of atypical genital bleeding. Nine months earlier, she had undergone a modified radical mastectomy for cancer of her right breast, estrogen receptor positive stage I (T1N0M0). The administration of tamoxifen, 20 mg/day, was started immediately postoperatively. Pelvic examination after tamoxifen administration for 9 months revealed that the uterus was enlarged to the size of a fist. Transvaginal ultrasonography and magnetic resonance imaging revealed a large solid mass with multiple cystic areas in the uterine cavity. The pathological diagnosis of the tumor after total hysterectomy was typical adenomyomatous polyp. It was believed to have developed during tamoxifen administration. PMID- 9390217 TI - Coxalgia as the initial symptom in Hodgkin's disease: a case report. AB - Primary Hodgkin's disease in the bone is extremely rare. We report the case of a 41-year-old woman with Hodgkin's disease, who had complained of left coxalgia 17 months prior to nodal involvement becoming evident clinically. She received combination chemotherapy with doxorubicin, bleomycin, vincristine and dacarbazine as well as radiotherapy to the pelvic lesion. Although the lymphadenopathy responded well to this treatment, the bone lesion was never in remission. The large mass of the bone lesion and its pelvic origin may explain the poor response to the cytotoxic therapies this patient received. The 22 cases reviewed showed that: 1, bony pain was the most frequent initial symptom; 2, nodal disease appeared in their clinical course in most cases; 3, the bones most commonly involved were pelvis, femur or tibia, and spine. PMID- 9390218 TI - Localized muscular mucormycosis in a child with acute leukemia. AB - Mucormycosis is a rare fungal infection of childhood, occurring mainly in patients with chronic illnesses such as diabetes and malignancies. The fungus seldom grows in culture and confirmation of the diagnosis depends on histologic examination of infected tissues. To date, the reported natural history of the disease has been rapid progression and a fatal outcome. Therefore, the importance of early diagnosis by tissue biopsy and early treatment with surgical debridement and systemic antifungal therapy cannot be overemphasized. The pulmonary system is the most common site for mucormycosis in patients with leukemia. We report what we believe to be the first successfully treated case of isolated muscular mucormycosis occurring in a child with biphenotypic acute leukemia. The diagnosis was made promptly by tissue examination at the time of surgical debridement. The patient was also given systemic amphotericin-B therapy. PMID- 9390219 TI - Report of the Tenth International Symposium of the Foundation for Promotion of Cancer Research: Basic and Clinical Research in Head and Neck cancer. PMID- 9390220 TI - Japanese challenge to gastric cancer surgeons: add 'meticulous accuracy' to Halsted's 'gentleness, haemostasis and minimal trauma'. PMID- 9390221 TI - Protein structure comparison using representation by line segment sequences. AB - This paper proposes a new comparison method of tertiary protein structures. The method consists of two parts. First, a sequence of line segments which approximates each tertiary protein structure is computed. Then, an alignment of the sequences of line segments corresponding to input structures is computed. The proposed method is considered as an intermediate one between two comparison methods: a method based on topology diagram and a method based on structure alignment. Moreover it takes less CPU time than structure alignment methods because most structures are represented by means of sequences of at most 100 line segments. The effectiveness of the method is confirmed through a comparison with a previous method and an application to database searching for similar structures. PMID- 9390222 TI - Quaternion contact ribbons: a new tool for visualizing intra- and intermolecular interactions in proteins. AB - Protein side chain interactions between residues separated by at least one loop or turn or break in the amino acid sequence are called 'nonlocal contacts' in this manuscript, and contiguous sets of such interactions located between segments of secondary structure are called 'contact zones.' A new interactive program, the quaternion contact ribbon tool, has been developed to help protein chemists identify, straighten if twisted, and display contact zones between two neighboring segments of helix. PMID- 9390223 TI - Fast protein fold recognition via sequence to structure alignment and contact capacity potentials. AB - We propose new empirical scoring potentials and associated alignment procedures for optimally aligning protein sequences to protein structures. The method has two main applications: first, the recognition of a plausible fold for a protein sequence of unknown structure out of a database of representative protein structures and, second, the improvement of sequence alignments by using structural information in order to find a better starting point for homology based modelling. The empirical scoring function is derived from an analysis of a nonredundant database of known structures by converting relative frequencies into pseudoenergies using a normalization according to the inverse Bolzmann law. These so called contact capacity-potentials turn out to be discriminative enough to detect structural folds in the absence of significant sequence similarity and at the same time simple enough to allow for a very fast optimization in an alignment procedure. PMID- 9390224 TI - A programming course in bioinformatics for computer and information science students. AB - We have created a course entitled "Representations and Algorithms for Computational Molecular Biology" with three specific goals in mind. First, we want to provide a technical introduction for computer science and medical information science students to the challenges of computing with molecular biology data, particularly the advantages of having easy access to real-world data sets. Second, we want to equip the students with the skills required of productive research assistants in molecular biology computing research projects. Finally, we want to provide a showcase for local investigators to describe their work in the context of a course that provide adequate background information. In order to achieve these goals, we have created a programming course, in which three major projects and six smaller assignments are assigned during the quarter. We stress fundamental representations and algorithms during the first part of the course in lectures given by the core faculty, and then have more focused lectures in which faculty research interests are highlighted. The course stressed issues of structural molecular biology, in order to better motivate the critical issues in sequence analysis. The culmination of the course was a challenge to the students to use a version of protein threading to predict which members of a set of unknown sequences were globins. The course was well received, and has been made a core requirement in the Medical Information Sciences program. PMID- 9390225 TI - Massively parallel algorithms for chromosome reconstruction. AB - Ordering clones from a genomic library into physical maps of whole chromosomes presents a central computational problem in genetics. Chromosome reconstruction via clone ordering is shown to be isomorphic to the NP-complete Optimal Linear Ordering problem. Massively parallel algorithms for simulated annealing based on Markov chain distribution are proposed and applied to this problem. Perturbation methods and problem-specific annealing heuristics are proposed and described. Experimental results on a 2048 processor MasPar MP-2 system are presented. Convergence, speedup and scalability characteristics of the various algorithms are analyzed and discussed. PMID- 9390226 TI - Structure formation of biopolymers is complex, their evolution may be simple. AB - Evolutionary strategies depend on the ability of evolving entities to conserve acquired features and to quickly adapt to new requirements as well. We use computer simulations of simplified exact biopolymers models to investigate the influence of mutations on structure formation. Our computations on large ensembles of random RNA secondary structures show that the sequence to structure mapping is ideally suited for evolutionary optimisation under point mutations: from any random structure it is not far to any target and yet most mutations will preserve the structure. The aim of this paper is to discuss the analogies as well as some recently developed methods to apply our approach to proteins: there we use Dill's HP - model of lattice proteins and apply a novel fast and efficient folding rule. There are remarkable similarities as both landscapes are rugged and structure formation largely depends on local interactions such that it is possible to accomplish a characterisation of the mapping similar to the RNA case. PMID- 9390227 TI - RNA pseudoknot modeling using intersections of stochastic context free grammars with applications to database search. AB - A model based on intersections of stochastic context free grammars is presented to allow for the modeling of RNA pseudoknot structures. The model runs relatively fast, having the same order running time as stochastic context free grammar parsers. The model is shown to be able to perform database searches and find RNA sequences which resemble RNA pseudoknots which bind biotin. The problem domain of RNA biotin binders has significance in the support of the RNA world model of early life on earth. PMID- 9390228 TI - How similar must a template protein be for homology modeling by side-chain packing methods? AB - Given the correct backbone coordinates of a globular protein, side-chain packing methods can be generally expected to predict the side-chain coordinates of the buried core residues accurately. In the context of a study in modeling a family of bacteriophage DNA-binding proteins, we observed that when the coordinates of the actual perfect backbone are not available, the side-chain packing methods are still of predictive value using homologous but imperfect backbones. This is the situation in practical homology modeling where a target protein sequence is modeled from template structures of known protein homologs. In order to assess the quality and degree of accuracy of such predictions and their dependence on the extent of homology, we have now extended these studies to a well characterized family of globin structures that span a much wider range of sequence-structure similarity. The collective results show a clear relationship that is independent of protein family between side-chain prediction accuracy and the level of similarity between the template and target proteins. We judge this similarity in terms of sequence identity and the backbone r.m.s. deviation of the template structure used for modeling and the actual target structure in cases where the target structures are available. In summary, as sequence identity drops from 100% to about 50%, or when the backbone r.m.s. deviation between template and target structures increases from 0 A to about 1 A, the overall average r.m.s. error for the buried-core residues rises from 1.2 A to 1.5 A while the chi 1 prediction accuracy drops from 85% to 70-75% and the chi 2 prediction accuracy drops from 80% to 60-65%. When the sequence identity drops below 50% or the backbone r.m.s. deviation rises above 1 A, all 3 measures of prediction accuracy decrease rapidly. When the sequence identity edges to the so-called twilight zone of sequence similarity at around 22%, or when the backbone r.m.s. deviation exceeds 2 A, the prediction accuracy approaches the values to be expected for random predictions, namely, 3.1 A for average r.m.s. error, 22% and 29% for accuracy of chi 1 and chi 2 prediction. These observations provide a practical evaluation of the side-chain packing methods and are of value to the homology modeler. The extent and degree to which the backbone topology of a protein fold can constrain internal side-chain orientation gives insight into the plasticity of the sequence-structure relationship found in the architecture of proteins. PMID- 9390229 TI - Mixed direct-iterative methods for boundary integral formulations of dielectric solvation models. AB - This paper describes a mixed direct-iterative method for boundary integral formulations of dielectric solvation models. We give an example for which a direct solution at thermal accuracy is nontrivial and for which Gauss-Seidel iteration diverges in rare but reproducible cases. This difficulty is analyzed by obtaining the eigenvalues and the spectral radius of the iteration matrix. This establishes that the nonconvergence is due to inaccuracies of the asymptotic approximations for the matrix elements for accidentally close boundary element pairs on different spheres. This difficulty is cured by checking for boundary element pairs closer than the typical spatial extent of the boundary elements and for those pairs performing an 'in-line' Monte Carlo integration to evaluate the required matrix elements. This difficulty are not expected and have not been observed when only a direct solution is sought. Finally, we give an example application of these methods to deprotonation of monosilicic acid in water. PMID- 9390230 TI - (Probably) all possible protein folds at low resolution. AB - For decades, a large number of investigators have been sifting the database of experimentally determined three-dimensional protein structures to discover recurring patterns of all types. Now that there are over a thousand such structures available, the natural question is whether we have seen all substantially different protein folds, and if not, how many have yet to be discovered? Answering the question can be broken down into three steps: (1) choose the range and domain for a similarity function, then (2) choose a particular similarity function, and (3) construct a corresponding protein model space that can be searched for dissimilar structures. In our analysis of the problem, we first chose to examine different conformations of the same protein, taking into account only C alpha atomic coordinates. In particular, we do not compare proteins of different chain lengths on the basis of some kind of gapped alignment. Secondly, we use a measure of conformational similarity based on rigid body superposition that emphasizes overall geometric resemblance, rather than agreement in secondary structure, for example. Third, we employed the discrete cosine transform to construct exhaustive sets of globular self-avoiding C alpha traces that were all different from each other by a given level. These sets of artificial structures were not too large to explicitly enumerate as long as the level of dissimilarity was high, and the chain flexibility was low. For chains flexible enough to match all experimental structures of 170 residue or less that are not beta-barrels, we find 128 artificial structures, of which 28 resemble nothing in the Protein Data Bank. PMID- 9390231 TI - DNA computing based on splicing: universality results. AB - The paper extends some of the most recently obtained results on the computational universality of specific variants of H systems (e.g. with regular sets of rules) and proves that we can construct universal computers based on various types of H systems with a finite set of splicing rules as well as a finite set of axioms, i.e. we show the theoretical possibility to design programmable universal DNA computers based on the splicing operation. For H systems working in the multiset style (where the numbers of copies of all available strings are counted) we elaborate how a Turing machine computing a partial recursive function can be simulated by an equivalent H system computing the same function; in that way, from a universal Turning machine we obtain a universal H system. Considering H systems as language generating devices we have to add various simple control mechanisms (checking the presence/absence of certain symbols in the spliced strings) to systems with a finite set of splicing rules as well as with a finite set of axioms in order to obtain the full computational power, i.e. to get a characterization of the family of recursively enumerable languages. We also introduce test tube systems, where several H systems work in parallel in their tubes and from time to time the contents of each tube are redistributed to all tubes according to certain separation conditions. By the construction of universal test tube systems we show that also such systems could serve as the theoretical basis for the development of biological (DNA) computers. PMID- 9390232 TI - Parallel discrete event simulation of Lyme disease. AB - Our research concerns the dynamic processes underlying the rapid increase in the geographic distribution of Lyme disease, currently the most frequently reported vector-borne disease of humans in the United States [10, 1]. More specifically, we ask how spatially localized ecological interactions drive the Lyme disease epidemic at extended spatial and temporal scales. We have developed a parallel discrete event simulation system in C++ for the IBM SP2. The simulation model discussed here models the mouse-tick interaction, an essential element of the epidemic's ecology. The main entities of the simulation are ticks in various stages of development (larval, nymphal, and adult) and mice. We track the behavior of mice and the spread of disease over the course of 180 days (late spring, summer, and early fall). Our goal is to understand patterns in the Lyme disease epidemic at the regional scale through studying the spread of the pathogen across a single white-footed mouse deme. PMID- 9390233 TI - Distance education through the Internet: the GNA-VSNS biocomputing course. AB - A prototype course on biocomputing was delivered via international computer networks in early summer 1995. The course lasted 11 weeks, and was offered free of charge. It was organized by the BioComputing Division of the Virtual School of Natural Sciences, which is a member school of the Globewide Network Academy. It brought together 34 students and 7 instructors from all over the world, and covered the basics of sequence analysis. Five authors from Germany and USA prepared a hypertext book which was discussed in weekly study sessions that took place in a virtual classroom at the BioMOO electronic conferencing system. The course aimed at students with backgrounds in molecular biology, biomedicine or computer science, complementing and extending their skills with an interdisciplinary curriculum. Special emphasis was placed on the use of Internet resources, and the development of new teaching tools. The hypertext book includes direct links to sequence analysis and databank search services on the Internet. A tool for the interactive visualization of unit-cost pairwise sequence alignment was developed for the course. All course material will stay accessible at the World Wide Web address (Uniform Resource Locator) http://+www.techfak.uni bielefeld.de/bcd/welcome .html. This paper describes the aims and organization of the course, and gives a preliminary account of this novel experience in distance education. PMID- 9390234 TI - Multifractals, encoded walks and the ergodicity of protein sequences. AB - A variety of statistical methods have been developed to explore correlations in protein and nucleic acid sequences. Such correlations have important implications for the evolution and stability of these macromolecules. Recently, a number of fractal analyses of sequence data have been developed. These analyses have considerable appeal as they are extremely sensitive to long range correlations and to hierarchical structures. One such analysis decodes sequence information into a random walk and the statistics of the resulting random walk is investigated. Anomalous scaling of such walks has been interpreted as indicative of a fractal structure. Alternatively, a generalized box counting analysis of decoded sequences can be used to establish multifractal properties. In this work, the connection between these two seemingly disparate approaches is established. This connection is exploited to investigate correlations in protein sequences. An ensemble consisting of a comprehensive data set of representative protein sequences is analyzed to establish the ergodicity of protein sequences. The implications of this ergodicity for information theoretical approaches to protein structure prediction is explored. PMID- 9390235 TI - List update processing (LUP)--solving the sequence database update problem. AB - Sequence databases of today require frequent updating. Mirror procedures to copy incrementally updated databases as cumulative sets are the preferred method and can be implemented by straightforward scripting. However, limited bandwidth of networks and the increase of data require more powerful paradigms to reduce the workload reliably. We suggest the List Update Processing (LUP) principle. The system has been implemented on an experimental basis to update the Swiss EMBnet Node (BioComputing Basel, CH) with data from the European Bioinformatics Institute (EMBL Outstation, Hinxton Hall, UK). The results obtained from the prototype suggest to expand the system to several sites. PMID- 9390236 TI - Circular clustering of protein dihedral angles by Minimum Message Length. AB - Early work on proteins identified the existence of helices and extended sheets in protein secondary structures, a high-level classification which remains popular today. Using the Snob program for information-theoretic Minimum Message Length (MML) classification, we are able to take the protein dihedral angles as determined by X-ray crystallography, and cluster sets of dihedral angles into groups. Previous work by Hunter and States has applied a similar Bayesian classification method, AutoClass, to protein data with site position represented by 3 Cartesian co-ordinates for each of the alpha-Carbon, beta-Carbon and Nitrogen, totalling 9 co-ordinates. By using the von Mises circular distribution in the Snob program, we are instead able to represent local site properties by the two dihedral angles, phi and psi. Since each site can be modelled as having 2 degrees of freedom, this orientation-invariant dihedral angle representation of the data is more compact than that of nine highly-correlated Cartesian co ordinates. Using the information-theoretic message length concepts discussed in the paper, such a more concise model is more likely to represent the underlying generating process from which the data came. We report on the results of our classification, plotting the classes in (phi, psi) space; and introducing a symmetric information-theoretic distance measure to build a minimum spanning tree between the classes. We also give a transition matrix between the classes and note the existence of three classes in the region phi approximately -1.09 rad and psi approximately -0.75 rad which are close on the spanning tree and have high inter-transition probabilities. This gives rise to a tight, abundant and self perpetuating structure. PMID- 9390237 TI - An object-oriented data-driven migration model. AB - This research uses object-oriented simulation to derive the migratory patterns of animal populations. These patterns are then compared with the trajectories of aquatic pollutant releases to yield quantitative biological impact assessments. These estimates are more realistic than those based on assumptions of uniform or random distributions of animals in a region--a common approach to this problem. The general migration model (MIGMOD) employs stochastic movement routines driven by biological field data to derive heterogenous migration patterns similar to those observed in nature. This data-driven approach circumvents many of the difficulties involved in migratory modelling from first principles. However, we also discuss the importance of selected causal mechanisms in these models. PMID- 9390238 TI - On the definition and the construction of pockets in macromolecules. AB - The shape of a protein is important for its functions. This includes the location and size of identifiable regions in its complement space. We formally define pockets as regions in the complement with limited accessibility from the outside. Pockets can be efficiently constructed by an algorithm based on alpha complexes. The algorithm is implemented and applied to proteins with known three-dimensional conformations. PMID- 9390239 TI - DNA splicing systems and post systems. AB - This paper concerns the formal study on the generative powers of extended splicing (H) systems. First, using a classical result by Post which characterizes the recursively enumerable languages in terms of his Post Normal systems, we establish several new characterizations of extended H systems which not only allow us to have very simple alternative proof methods for the previous results mentioned above, but also give a new insight into the relationships between families of extended H systems. We show a kind of normal form for extended H systems exactly characterizing the class of regular languages. We also show a new representation result for the family of context-free languages in terms of extended H systems. PMID- 9390240 TI - Assessing the performance of fold recognition methods by means of a comprehensive benchmark. AB - Recently there has been an explosion of methods for fold recognition. These methods seek to align a protein sequence to a three-dimensional structure and measure the compatibility of the sequence to the structure. In this work, we present a benchmark to assess the performance of such methods. The benchmark consists of a set of protein sequences matched by superposition to known structures. This set covers a wide range of protein families, and includes matching proteins with insignificant sequence similarity. To demonstrate the usefulness of this benchmark, we apply it here to compare different fold recognition methods developed through the years in our group as well as several sequence-sequence substitution matrices. The results show that "global-local" alignments are superior to either local or global alignments. The most effective sequence-sequence matching matrix is the Gonnet table. The best performance overall is obtained by a method which combines the 3D-1D profiles of Bowie et al. with a substitution matrix and takes into account residue pairwise interactions. PMID- 9390241 TI - Biocomputing education by the Australian National Genomic Information Service. PMID- 9390242 TI - Puzzle pieces defined: locating common packing units in tertiary protein contacts. AB - Puzzle pieces are defined as small packing units which make up the unique tertiary interactions in proteins. Anti-parallel and perpendicular helix-helix contacts were broken down into basic puzzle-piece pairs in order to study the traits of such contacts: their limited geometry, preferred residue involvement, residue conformation and other common constraints. These traits can then be used for continued comparison of other protein structures, improving models of and designing proteins de novo and, in time, predicting 3D structure from primary sequence. Results from a small (100 proteins) database of anti-parallel helix helix contacts and from preliminary work on a large database (600 proteins) of perpendicular helix-helix contacts are presented. PMID- 9390243 TI - Using multiple alignments and phylogenetic trees to detect RNA secondary structure. AB - We describe a statistical method to determine if a pair of columns in a multiple alignment of a homologous family of RNA sequences shows evidence of being base paired. The method makes explicit use of a given phylogenetic tree for the sequences in the alignment. It is tested on a multiple alignment of 16S rRNA sequences with good results. PMID- 9390244 TI - A branch and bound algorithm for local multiple alignment. AB - A Branch and Bound Algorithm has been developed to find a set of window positions in a compilation of sequences with globally maximal information content. We have also developed an algorithm for brute force evaluation of solutions which is faster by a factor of the length of the windows than the naive brute force algorithm. The combination of these two algorithms allows us to solve problems to optimality that were previously amenable only to heuristic algorithms. PMID- 9390245 TI - Beyond the hyperactive molecule: search, salvage and visualization of chemical information from the Internet. AB - The established exchange mechanisms for chemical information are under attack from new information distribution channels on the Internet. Increasingly chemical information is distributed by means of WWW pages and similar media. However, most of this information is still primarily intended for human browsing. The search for chemical information and the reuse of encoded structures and their attached data is complicated and often impossible because of the unorganized structure of the information and the lack of tools for search, display and salvage of chemical information to help with the extraction of reusable information from Webspace. The situation is complicated by the lack of standards and formats powerful enough to encode in computer-readable form sophisticated chemical information and informational relationships. The rapid evolution of information exchange mechanisms on the Internet is another problem. The unsettled situation demands a new generation of intelligent chemistry-aware tools for information retrieval from the Net. These tools must be capable of adapting to new trends and information models as well as new information types without constant redesign and should be themselves extendable and updatable by components distributed via the same network connections as the chemical data they are supposed to deal with. We introduce a set of tools which encapsulate the established Internet (especially WWW) information transfer and visualization methods and extend them to a new level of chemical information handling. PMID- 9390246 TI - An introductory course in computation molecular biology: rationale, history, observations, and course description. AB - A course called "Molecular Biology Computer Techniques" was implemented in 1987 and has been evolving ever since. Currently the semester-long three credit course consists of thirty hours of lecture (three hours/week for the first ten weeks of the semester) and a minimum of 45 hours of laboratory instruction (three hours/week). The lectures survey both bioinformatics and structure based methods. The laboratory has two tracks, one that can be described loosely as "sequence analysis" and the other as "molecular modelling." Most students choose one of the two laboratory tracks, although a small number have done both, either simultaneously or in successive years. For each student, the goal of the course is the completion of a student-initiated research project. The culmination of the course is the presentation of the completed projects at a "Poster Session Final." During this final, which is conducted like a poster session at a typical biological science meeting, students are examined, not only by the instructors in the course, but also by a diverse cross-section of the university community at large, including non-scientists (who are specially invited to attend). Questioning by non-scientists provides opportunity for the students to improve their communication skills with the lay public. In this manuscript we discuss our views regarding the rationale for the development of formal courses in computational molecular biology, relate our experiences in the development of our course, and describe the course as it stood the last time it was taught, which was in the Fall of 1994. PMID- 9390247 TI - Visualisation in the SPROUT molecular design program. AB - SPROUT is an interactive computer system for structure based molecular design. The system consists of several modules that address the different subproblems of structure based drug design. This paper describes the visualisation techniques applied in the program: the display of the novel (geometric region) representation of the interaction sites and the molecular surface display based on a 3D grid representation of the cavity. The hydrogen bonding regions are represented by set operations (subtraction and intersection) of simple spherical and conical 3D objects (with given radii and opening angle) Some complex hydrogen bonding regions are represented by intersections of six or more basic objects. A method for calculating a triangular mesh representation (with normal vectors) of the analytical surfaces of the objects, that have sharp edges and corners because of the intersections, is presented in the paper. The geometric parameters of the interaction regions can be changed interactively in which case the surface display is updated real-time. The volume of space that is available for ligand generation (the cavity of the receptor site) is represented on a 3D grid within SPROUT. The surface of the available space is visualised using an algorithm presented in the paper, that generates a polygonial mesh of the grid points. The grid is also used to cut out stericaly forbidden parts of the interaction site regions. The surface of the reduced object is also visualised using further sphere subtractions. The presented algorithms are fast, aplicable in interactive visualisation programs. Result images of the rendering of the surfaces, calculated by the algorithms, are demonstrated on examples taken from applications of SPROUT to practical ligand design problems. PMID- 9390248 TI - Computational evolution of a model polymer that folds to a specified target conformation. AB - A method is described for folding polymers to specific target conformations. The approach uses a fast but approximate dynamics algorithm, coupled with a genetic algorithm that is used to evolve the large number of free parameters needed. The dynamics algorithm uses a state transition matrix approach. At each time step, the distances between pairs of atoms are adjusted by shifting them from Dij to Dij + Sij where Sij is an element of the state transition matrix S. Atom pairs that are attractive have Sij < 0 and pairs that are repulsive have S ij > 0. The atomic movement is carried out by gradient minimizing the molecular mechanics energy of the molecule subject to harmonic distance constraints. The method is applied to a simple test case, a 19 atom 2-D polymer. The paper also show that the S matrices can correctly fold a limited variety of initial conformations that differ from the one used during the evolution phase. PMID- 9390249 TI - A protocol for maintaining multidatabase referential integrity. AB - The bioinformatics community is becoming increasingly reliant on the creation of links among biological databases (DBs) as a foundation for DB interoperability. For example, a link might be created from a protein in one DB (such as PIR), to a gene in another DB (such as GDB), by storing the unique identifier (id) of the gene object within an attribute of the protein object. User interfaces can then support navigation from the protein to the gene, and multiDB queries can join the protein with the gene. The unique id of the gene is serving as a foreign key. However, a variety of factors, such as changes in the underlying biology, can cause object ids to become invalid, thus producing invalid links among DBs. Invalid links are a violation of multidatabase referential integrity. We propose a network protocol whereby a database administrator can provide information about changes to the identifiers of objects in their database via Internet, to allow other databases to maintain referential integrity. We request comments from the bioinformatics community for the purpose of building a consensus on the proposed protocol. PMID- 9390250 TI - An algorithm for prediction of structural elements in small proteins. AB - A method for predicting the location of surface loops/turns and assigning the intervening secondary structure of the transglobular linkers in small, single domain globular proteins has been developed. Application to a set of 10 proteins of known structure indicates a high level of accuracy. The secondary structure assignment in the center of transglobular connections is correct in more than 85% of the cases. A similar error rate is found for loops. Since more global information about the fold is provided, it is complementary to standard secondary structure prediction approaches. Consequently, it may be useful in early stages of tertiary structure prediction when establishment of the structural class and possible folding topologies is of interest. PMID- 9390251 TI - Sequence sizes of eukaryotic enzymes. AB - We have shown in earlier studies that an appreciable fraction of proteins display sequence size periodicity with periods of approximately 123 aa and approximately 152 aa for eukaryotes and prokaryotes, respectively. For any firm conclusions to be made, the issue of possible bias due to an overabundance of some protein families should be addressed in more than one way. Here we present the size distributions for various sequence ensembles of eukaryotic enzymes that differ by level of data bank cleaning. The sequences were purged by applying several successive thresholds of relatedness irrespective of the sequence lengths. The previously observed preference to typical sizes is confirmed. Possible reasons for the observed excess of the typical size sequences are discussed. PMID- 9390252 TI - A high performance system for molecular dynamics simulation of biomolecules using a special-purpose computer. AB - GRAPE (GRavity PipE) processors are special purpose computers for simulation of classical particles. The performance of MD-GRAPE, one of the GRAPEs developed for molecular dynamics, was investigated. The effective speed of MD-GRAPE was equivalent to approximately 6 Gflops. The precision of MD-GRAPE was good judging from the acceptable fluctuation of the total energy. Then a software named PEACH (Program for Energetic Analysis of bioCHemical molecules) was developed for molecular dynamics of biomolecules in combination with MD-GRAPE. Molecular dynamics simulation was performed for several protein-solvent systems with different sizes. Simulation of the largest system investigated (27,000 atoms) took only 5 sec/step. Thus, the PEACH-GRAPE system is expected to be useful in accurate and reliable simulation of large biomolecules. PMID- 9390253 TI - Correlating structure-dependent mutation matrices with physical-chemical properties. AB - We have investigated how structure-dependent mutation matrices derived in previous work correlate with various physical-chemical properties of the 20 naturally occurring amino acids. Among the properties we investigated were delta G of transfer from water to octanol and cyclohexane, alpha helical and beta sheet propensity, size, and charge. We found that the delta G of transfer to octanol had a high correlation with matrices for all categories of residues, especially the matrices for buried and exposed positions. This result suggests that octanol is a good model for understanding both the changes in stability resulting from substitutions of buried residues and changes in foldability resulting from varying exposed residues. We also found the correlations of the matrices with size and charge varied with the local environment, and that neither alpha helical nor beta sheet propensity had high correlations with most matrices. Thus, conservation of size and charge appear to be important in specific environments, and conservation of alpha helix and beta sheet propensity do not seem to be key factors. PMID- 9390254 TI - A case study where biology inspired a solution to a computer science problem. AB - This paper describes how the biological theory of gene duplication described in Susumu Ohno's provocative book, Evolution by Means of Gene Duplication, was brought to bear on a vexatious problem from the domain of automated machine learning, namely the problem of architecture discovery. Six new architecture altering operations for genetic programming were motivated by the way that new biological structures, functions, and behaviors arise in nature using gene duplication. Genetic programming with the new architecture-altering operations was then applied to the transmembrane protein segment identification problem. The out-of-sample error rate for the best genetically-evolved program achieved was slightly better than that of previously-reported human-written algorithms for this problem. PMID- 9390255 TI - Protein phylogenetic inference using maximum likelihood with a genetic algorithm. AB - This paper presents a method to construct phylogenetic trees from amino acid sequences. Our method uses a maximum likelihood approach which gives a confidence score for each possible alternative tree. Based on this approach, we developed a genetic algorithm for exploring the best score tree: randomly generated alternative trees are rearranged so that their scores are improved by utilizing crossover and mutation operators. In a test of our algorithm on a data set of EF 1 alpha sequences, we found that the performance of our algorithm is comparable to that of other tree-construction methods. PMID- 9390256 TI - Ab initio calculations of free energy barriers for chemical reactions in solution: proton transfer in [FHF]-. AB - This paper describes a hybrid ab initio quantum mechanical/molecular mechanics (QM/MM) method for calculating activation free energies of chemical reactions in solution, using molecular mechanics force fields for the solvent and an ab initio technique that incorporates the potential from the solvent in its Hamiltonian for the solute. The empirical valence bond (EVB) method is used as a reference potential for the ab initio free energy calculation, and drives the reaction along the proper coordinate, thus overcoming problems encountered by direct attempts to use molecular orbital methods in calculations of activation free energies. The utility of our method is illustrated by calculating the activation free energy for proton transfer between fluoride ions in the [FHF]-system, in both polar and nonpolar solution. PMID- 9390257 TI - Mixed quantum mechanical/molecular mechanical simulations of chemical reactions in solution and in enzymes by the classical trajectory mapping approach. AB - We present a practical hybrid quantum mechanical/molecular mechanical approach to study chemical reactions in solution and in enzymes. In this method, referred to as the "Classical Trajectory Mapping" method, trajectories are calculated on the classical potential surfaces and, by using the classical surfaces as a reference state for the actual quantum mechanical ground state potential, the free energy profile of the chemical reaction is obtained by the free energy perturbation technique. This method was applied to proton-transfer reactions both in aqueous solution and in papain. The encouraging results indicate the applicability of our method to chemical reactions in the condensed phase and the biological systems. PMID- 9390258 TI - Molecular modeling of protocellular functions. AB - The mechanisms of three protocellular functions have been studied using molecular modeling techniques. These functions are (1) the transport of ions across membranes, (2) the formation of photoactivated proton gradient that could drive chemical synthesis in the protocell, and (3) the organization of small peptides necessary for catalytic activity. In all these processes, membranes play an essential role. The transfer of ions across the barrier formed by protocellular walls is facilitated by the formation of deep, thinning defects in the membrane. Membranes also form a barrier to charged species that allows for retaining proton gradients. These gradients can be generated by a simple transmembrane proton pump consisting of a proton source and two acceptors. The directionality of the pump is ensured by a "gate-keeping" mechanism involving a water molecule, conformational change of the primary acceptor or tautomerization of a histidine. The pump can be formed by two transmembrane helices but not one helix. They provide surfaces at which organic molecules concentrate and small peptides can organize into ordered, amphiphilic structures. In general, valuable information about the origins and evolution of protocells can be obtained from the knowledge of physical and chemical principles that govern functioning of contemporary cells. PMID- 9390259 TI - Analysis, clustering and prediction of the conformation of short and medium size loops connecting regular secondary structures. AB - Loops are regions of non-repetitive conformation connecting regular secondary structures. They are both the most difficult and error prone regions of a protein to solve by X-ray crystallography and the hardest regions to model using knowledge-based procedures. While the core of a protein can be straight forwardly modelled from the structurally conserved regions of homologues of known structure, loops must be modelled from a selected homologue or from a loop chosen from outside the family. Here we present a loop prediction procedure that attempts to identify the conformational class of the loop rather than to select a specific loop from a database of fragments. The structures of some 2083 loops of one to eight residues in length were extracted from a database of 225 protein and protein domain structures. For each loop, the relative disposition of its bounding secondary structures is described by the separation between the tips of their axes, the angle and dihedral angle between their axes. From the clustering of the loops according to the root mean square deviation of their spatial fit, a total of 162 loop conformational classes, including 79% of loops, were identified. One-hundred and eight of these, involving 66% of the loops, were populated by at least four non-homologous loops or four loops sharing a low sequence identity. Another 54 classes, including 13% of the loops, were populated by at least three loops of low sequence similarity from three or fewer non homologous groups. Most of the previously described loop conformations were found among the populated classes. For each class a template was constructed containing both sequence preferences and the relative disposition of bounding secondary structures among member loops. During comparative modelling, the conformation of a loop can be predicted by identifying a loop class with which its sequence and disposition of bounding secondary structures are compatible. PMID- 9390260 TI - Computational biology instruction at the University of Washington Center for Bioengineering. AB - Computational tools are rapidly becoming an essential component of molecular biology research. However, there is as yet relatively little attention paid to computational biology in the standard curricula of most biology programs. We describe here some of the graduate computational biology courses we have developed in the Center for Bioengineering at the University of Washington, with special emphasis on instructional methods and approaches that appear to work well. PMID- 9390261 TI - A greedy strategy for finding motifs from yes-no examples. AB - We define a motif as an expression Z1.Z2...Zn with sets Z1, Z2,..., Zn of strings in a specified family omega called the type. This notion can capture the most of the motifs in PROSITE as well as regular pattern languages. A greedy strategy is developed for finding such motifs with ambiguity just from positive and negative examples by exploiting the probabilistic argument. This paper concentrates on describing the idea of the greedy algorithm with its underlying theory. Its experimental results on splicing sites and E. coli promoters are also presented. PMID- 9390262 TI - Statistical geometry analysis of proteins: implications for inverted structure prediction. AB - The topology of folded proteins from the representative dataset of well-defined three-dimensional protein structures is studied using a statistical geometry approach. Amino acid residues in protein chains are represented by C alpha atoms, thus reducing the protein three-dimensional structure to a set of points in three dimensional space. The Delaunay tessellation of a protein structure generates an aggregate of space-filling irregular tetrahedra, or Delaunay simplices. Each simplex objectively defines four nearest neighbor C alpha atoms, i.e. four nearest neighbor residues. The statistical analysis of residue composition of Delaunay simplices reveals nonrandom preferences for certain quadruplets of amino acids. These nonrandom preferences are used to develop a fitness function that evaluates sequence-structure compatibility. Using this fitness function, several tested native proteins score the highest among 100,000 random sequences with average protein amino acid composition. The statistical geometry approach, based solely on first principles, provides a unique means for protein structure analysis and has direct implications for inverted protein structure prediction. PMID- 9390263 TI - Determination of proton transfer rate constants using Ab initio, molecular dynamics and density matrix evolution calculations. AB - In this work we give an overview of the methodologies required to compute the rate of proton transfer in hydrogen bonded systems in solution. Using ab initio or density functional methods we determine proton potentials of a truncated system as a function of proton-donor proton-acceptor distance as well as nonbonding parameters. By classical molecular dynamics we evaluate a swarm of proton potentials with the proton fixed in the reactant well. The rate of proton transfer is calculated perturbatively using the Density Matrix Evolution (DME) method, going beyond the Born Oppenheimer approximation. The method is illustrated by two examples: hydrogen malonate and the active center of HIV-1 protease. PMID- 9390264 TI - Empirical free energy calculations of human immunodeficiency virus type 1 protease crystallographic complexes. II. Knowledge-based ligand-protein interaction potentials applied to thermodynamic analysis of hydrophobic mutations. AB - Empirical free energy calculations of HIV-1 protease crystallographic complexes based on the developed knowledge-based ligand-protein interaction potentials have enabled a detailed thermodynamic analysis. Binding free energies are estimated within an empirical model that postulates that hydrophobic effect, mean field ligand-protein interaction potentials and conformational entropy changes are the dominant forces that determine complex formation. To provide a quantitative framework of the binding thermodynamics contributions the derived knowledge-based potentials have been linked with the hydrophobicity and conformational entropy scales originally developed to explain protein stability. The comparative analysis of studied inhibitors provides reasonable estimates of distinctions in their binding affinity with HIV-1 protease and gives insight into the nature of the binding determinants. The binding free energy changes upon a simple hydrophobic mutation Ile -> Val in the JG-365, MVT-101 and U75875 inhibitors of HIV-1 protease have been evaluated within a model that includes the effects of solvation, cavity formation, conformational entropy and mean field ligand-protein interactions. In general, free energy changes associated with a particular perturbation of a system can not be rigorously decomposed into separate terms from first principles. We explored the relationships between the changes in hydrophobic contributions and mean field ligand-protein interaction energies in the context of a totally buried and dense area of the binding site. We assume, therefore, that these simple hydrophobic deletions would not induce noticeable conformational changes in the enzyme and can be interpreted with some confidence in the framework of the model. The analysis has revealed the decisive effect of the energetics of ligand-protein interactions on the estimated free energy changes. PMID- 9390265 TI - Prediction of the quaternary structure of coiled coils: GCN4 leucine zipper and its mutants. AB - A methodology for predicting coiled coil quaternary structure and for the dissection of the interactions responsible for the global fold is described. Application is made to the equilibrium between different oligomeric species of the wild type GCN4 leucine zipper and seven of its mutants that were studied by Harbury et al. Over the entire experimental concentration range, agreement with experiment is found in five cases, while in two other cases, agreement is found over a portion of the concentration range. These simulations suggest that the degree of chain association is determined by the balance between specific side chain packing preferences and the entropy reduction associated with side chain burial in higher order multimers. PMID- 9390266 TI - 3D molecular graphics on the World Wide Web. AB - The Virtual Reality Modeling Language (VRML) is a new tool for the information transfer on the World Wide Web (WWW). We present a short description and some applications in the field of molecular graphics of this new, object oriented language. It is shown that the usage of VRML is a very efficient and powerful method for the transport of molecular models over the net. PMID- 9390267 TI - Motif identification neural design for rapid and sensitive protein family search. AB - The accelerated growth of the molecular sequencing data has generated a pressing need for advanced sequence annotation tools. This paper reports a new method, termed MOTIFIND (Motif Identification Neural Design), for rapid and sensitive protein family identification. The method is extended from our previous gene classification artificial neural system and employs two new designs to enhance the detection of distant relationships. These include an n-gram term weighting algorithm for extracting local motif patterns, and integrated neural networks for combining global and local sequence information. The system has been tested with three protein families of electron transferases, namely cytochrome c, cytochrome b and flavodoxin, with a 100% sensitivity and more than 99.6% specificity. The accuracy of MOTIFIND is comparable to the BLAST database search method, but its speed is more than 20 times faster. The system is much more robust than the PROSITE search which is based on simple signature patterns. MOTIFIND also compares favorably with the BLIMPS search of BLOCKS in detecting fragmentary sequences lacking complete motif regions. The method has the potential to become a full-scale database search and sequence analysis tool. PMID- 9390268 TI - Extraction of hidden Markov model representations of signal patterns in DNA sequences. AB - We have developed a method to extract the signal patterns in DNA sequences. In this method, the Genetic Algorithm (GA) and Baum-Welch algorithm are used to obtain the best Hidden Markov Model (HMM) representations of the signal patterns in DNA sequences. The GA is used to search the best network shapes and the initial parameters of the HMMs. Baum-Welch algorithm is used to optimize the HMM parameters for the given network shapes. Akaike Information Criterion (AIC), which gives a criterion for the balance of adaptation and complexity of a model, is applied in the HMM evaluation. We have applied the method to the extraction of the signal patterns in human promoters and 5' ends of yeast introns. As a result, we obtained HMM representations of characteristic features in these sequences. To validate the efficiency of the method, we have performed promoter recognition using obtained HMMs. Two entries including nine promoters are selected from GenBank 76.0, and it is observed that the HMM can predicts eight promoters correctly. These results imply that the method is efficient to design preferable HMM networks, and provides reliable models for the recognition of the signal patterns. PMID- 9390269 TI - [Extending the capabilities of human chromosome analysis: from high-resolution banding to chromatin fiber-FISH]. AB - The rapid development in human chromosome studies during the past 2 decades has opened up a variety of new avenues in both basic and clinical human genetics. This has been due to the extended resolution of chromosome analysis especially by the introduction of high-resolution banding and fluorescence in situ hybridization (FISH) techniques. High-resolution banding methods using elongated chromosomes from cells at early mitotic stage can be divided into two main categories: one involves S-phase synchronization using methotrexate or thymidine with subsequent release from the block, and the other involves the application of DNA-binding agents, such as ethidium bromide, which inhibit mitotic chromosome condensation. High-resolution band analysis facilitates refined determination of the breakpoints on rearranged chromosomes, and thus minute deletions, small translocations and other subtle chromosome mutations can readily be detected and identified. It has also proved to be valuable for accurate gene mapping and comparative cytogenetics. Each of the high-resolution bands consists of DNA molecules as long as approximately 3Mb on the average, and this resolution limitation has been overcome by the introduction of the FISH method, which enables chromosome and genomic analyses to be carried out at the DNA molecule level. For FISH analysis, appropriate DNA probes of various insert sizes can be chosen for different purposes, and FISH with specific probes permits high resolution analysis of chromosome DNA constitution at particular loci. For instance, it is possible to detect and map unique sequences more than 1 kb in size on mitotic chromosomes, and to identify cryptic chromosome rearrangements not recognizable even by high-resolution band analysis. Better resolution of breakpoint determination can also be expected by FISH using an adequate series of DNA clones arrayed on the physical map within the relevant chromosome region. An even better genomic resolution by FISH can be obtained by utilizing free chromatin DNA fibers released from interphase nuclei as a hybrization target. The FISH signals on the chromatin fiber preparations are shown to be "linear", unlike the standard FISH signals that have a "dot" appearance, and the resolution attainable should be comparable to analysis of a straight DNA double helix itself. The degree of resolution in the so-called fiber-FISH method ranges from a few to 300 kb, and thus the method can readily be applied to the high-resolution assessment of ordering and overlapping of isolated DNA clones from a specific chromosome region, as well as sizing of gaps between the clones. Fiber-FISH may prove most valuable for the precise determination of breakpoints at the kilobase level, and also for identifying minute deletions and duplications. By presenting some examples of our own recent works, this review attempts to summarize the current contribution of the above strategies in mediating the two separate ranges of resolution achieved by the classical chromosome and molecular DNA analyses. PMID- 9390270 TI - Costimulation of human natural killer cell proliferation: role of accessory cytokines and cell contact-dependent signals. AB - Despite the importance of natural killer (NK) cells in the immune response, the regulation of human NK cell growth has not been well characterized. We have hypothesized that, similar to the proliferation of T and B lymphocytes, optimal proliferation of NK cells requires costimulatory signals as well as a primary mitogenic stimulus. Evidence for costimulation by both soluble cytokines and cell contact-dependent factors is presented. Soluble IL-1 and TNF were found to augment NK cell proliferation in response to primary mitogenic cytokines, including IL-2, IL-4, IL-7, and IL-12. The costimulatory effect of IL-1 and TNF is strongly enhanced by the calcium ionophore ionomycin. Coculture of NK cells with irradiated K562 cells can largely substitute for the costimulatory signal provided by ionomycin. Costimulation by K562 requires intimate cell contact and is not reconstituted by cell-free supernatants. Activated T lymphocytes can also mediate contact-dependent costimulation of NK cells; resting PBMC, several NK sensitive cell lines, and all NK-resistant cell lines tested were not found to be costimulatory. Engagement of CD16 did not augment NK cell proliferation. Thus, triggering of natural killing or antibody-dependent cell-mediated cytotoxicity (ADCC) does not consistently provide a costimulatory signal for NK cell proliferation. Cell contact-dependent costimulation of NK cells does not appear to involve known receptors that can costimulate T cells, including CD2, CD27, CD28, CD29, or LFA-1. The molecular nature of the putative NK cell costimulatory receptor remains to be elucidated. Nevertheless, human NK cells could be expanded in vitro using leukocyte-conditioned medium (LCM) as a source of IL-2 and accessory cytokines and ionomycin to bypass the putative receptor for cell contact-dependent costimulation. NK cells expanded in LCM and ionomycin express typical NK cell antigens and mediate natural killing and ADCC. Further characterization of the costimulatory signals for NK cell proliferation may elucidate the physiologic regulation of NK cell growth and may ultimately allow more effective manipulation of these lymphocytes in the immunotherapy of human diseases. PMID- 9390271 TI - Positive selection of CD56+ lymphocytes by magnetic cell sorting. AB - A new method for the isolation of CD56+ lymphocytes from peripheral mononuclear blood cells is described. Magnetic microbeads conjugated to goat antimouse antiserum in combination with a murine monoclonal anti-CD56 antibody were coated to the CD56+ target cells. CD56+ cells were then isolated with the use of a magnetic cell sorter. The purity of the CD56+ cells was 98.4 +/- 1% (n = 12) with a recovery of the CD56+ lymphocytes of 57.2 +/- 9% (range 48-77%). The natural killer, activity of the CD56+ lymphocytes as well as the interleukin-2-induced proliferative response were not affected by the isolation procedure or the presence of the magnetic microbeads on the CD56+ cells. The described method might be a useful tool for the further characterization of CD56+ cells and their subsets and can easily be upgraded for clinical use in adoptive immunotherapy with CD56+ lymphocytes. PMID- 9390272 TI - Mannan-binding protein in human umbilical cord blood. AB - Mannan binding protein (MBP) may be important for host defence particularly in infancy. MBP concentration was measured in 237 umbilical cord blood samples from singleton pregnancies and compared to those of 352 blood donors. Both data sets yielded a bimodal frequency distribution, consisting of a log-normal peak and a long tail of lower values. The range (0-23 U/ml) and median (7.2 U/ml) of cord blood values were significantly lower than those of blood donors (range 0-43 U/ml; median 8.3 U/ml). MBP was also measured in the cord blood samples of 8 pairs of twin siblings. Discordant values in two pairs of twins suggest that cord blood MBP is derived from the fetoplacental unit and not from the maternal circulation by transplacental passage. PMID- 9390273 TI - Natural resistance to Mycoplasma pulmonis infection in mice: host resistance gene(s) map to chromosome 4. AB - Strains of mice differ greatly in resistance to infection in their lungs with virulent Mycoplasma pulmonis (MP) organisms even during the first 5 days, prior to detection of humoral or T cell mediated acquired immune responses. C57BL/6 mice are resistant, and BALB/c and C3H mice are susceptible, and one major gene, MP, not linked to the H2 major histocompatibility complex, regulates resistance. C57BL/6 x C3H (B x H) and BALB/c x C57BL/6 (C x B) recombinant inbred strain mice were infected intratracheally with the T2 strain of MP. Five days later, the recovery of organisms from tracheolung lavages and lung tissue was determined. The strain distribution pattern of resistance indicated that the MP gene maps to chromosome 4. B6.C-H18 (B6 mice congenic for the BALB/c H18 gene of chromosome 4) were much more susceptible than B6 mice, but were less susceptible than BALB/c mice, supporting the data obtained with the recombinant inbred strain mice, but suggesting that other genes may also influence resistance to infection with MP. PMID- 9390274 TI - Exacerbation of toxoplasmosis in neutrophil-depleted mice. AB - Studies were performed to determine whether resistance to Toxoplasma gondii infection in mice depends on a mechanism involving neutrophils. Immunocompetent C57BL/6 and C.B-17 mice infected with T. gondii by gavage had an increased percentage of neutrophils in their peripheral blood. C57BL/6 mice selectively depleted of neutrophils by injections of RB6-8C5 monoclonal antibody died during the acute phase of the disease. Depletion of neutrophils had no effect on interferon gamma production, but had a profound effect on the total numbers of peripheral blood CD4+ and CD8+ T cells. Neutrophil-depleted C.B-17 mice survived longer than neutrophil-depleted C57BL/6 mice when infected with T. gondii, however they became much sicker, and were less able to survive long-term than infected, control mAb-treated mice as indicated by severe sustained weight loss. This study shows that neutrophils play an important role in resistance to acute primary T. gondii infection and that depletion of neutrophils reduces the numbers of CD4+ and CD8+ lymphocytes recoverable from peripheral blood of infected but not uninfected mice. This effect on lymphocytes may contribute to the reduced long-term survival of neutrophil-depleted mice. PMID- 9390276 TI - New challenges in computational biochemistry. PMID- 9390275 TI - NKR-P1dim/TCR alpha beta + T cells and natural killer cells share expression of NKR-P1A and NKR-P1D. AB - Natural killer (NK) cells and a T cell subset express NKR-P1s; however, it is not known if NKR-P1s on these cells are homologous. By molecular and biochemical means, we determined that NKR-P1s on NK cells and T cells were similar. Also, sequencing of polymerase chain reaction products derived from these populations indicated expression of a novel form, termed NKR-P1D, with approximately 60% nucleotide homology to NKR-P1A. NKR-P1D shares approximately 50% amino acid homology, overall and in the carbohydrate recognition domain, with rat NKR-P1A and mouse NKR-P1A -B, and -C. Transcripts for NKR-P1A (1.1 kb) and NKR-P1D (2.0 kb) were produced by NK cells and NKR-P1dim/TCR alpha beta + T cells. Some NK cell clones coexpressed NKR-P1A and NKR-P1D. However, other clones lacking NKR P1A produced message for NKR-P1D. PMID- 9390277 TI - Computer simulations of prebiotic evolution. AB - This paper is a review of our previous work on the field of possible ways of prebiotic evolution. We propose an algorithm providing sequences of model proteins with rapid folding into a given native conformation. Thermodynamical analysis shows that the increase in speed is matched by an increase in stability: the evolved sequences are much more stable in their native conformation than the initial random sequence. We discuss a possible origin of the first biopolymers, having stable unique structure. We suggest that at the prebiotic stage of evolution, long organic polymers had to be compact in order to avoid hydrolysis and had to be soluble and thus must not be exceedingly hydrophobic. We present an algorithm that generates such sequences of model proteins. The evolved sequences turn out to have a stable unique structure, into which they quickly fold. This result illustrates the idea that the unique three-dimensional native structure of first biopolymers could have evolved as a side effect of a nonspecific physico chemical factors acting at the prebiotic stage of evolution. PMID- 9390278 TI - Ubiquitous distributed objects with CORBA. AB - Database interoperation is becoming a bottleneck for the research community in biology. In this paper, we first discuss the question of interoperability and give a brief overview of CORBA. Then, an example is explained in some detail: a simple but realistic data bank of STSs is implemented. The Object Request Broker is the media for communication between an object server (the data bank) and a client (possibly a genome center). Since CORBA enables easy development of networked applications, we meant this paper to provide an incentive for the bioinformatics community to develop distributed objects. PMID- 9390279 TI - Towards a density functional treatment of chemical reactions in complex media. AB - We discuss two techniques involving density functional theory (i.e., ab initio molecular dynamics simulations and frozen density functional theory) which show promise for applications directed towards understanding reactions in complex media. Preliminary results for the simulation of the conformational dynamics of an isolated analogue of alanine dipeptide and for the interaction between F- and H2O are included. These results represent the first steps towards a combined theoretical approach of reactions in complex media. PMID- 9390280 TI - Combinatorial tools for the analysis of transcriptional regulation. AB - In this paper, we discuss virtual experiments for the study of major regulatory processes such as translation, signalization or transcription pathways. An essential part of these processes is the formation of protein clusters held together by a small number of binding domains that can be shared by many different proteins. Analysis of these clusters is complicated by the vast number of different arrangements of proteins that can trigger a specific reaction. We propose combinatorial tools that can help predict the effects on the rate of transcription of either changes in transcriptional factors concentration, or due to the introduction of chimeras combining domains not usually present on a protein. PMID- 9390281 TI - The generalization method of relationships among nucleotide sequences reveals an order in assimilation of amino acid codons during the isoacceptor tRNAs evolution. AB - A new method, generalization of relationships among the nucleotide sequences has been used to analyze divergency both within and across the families of isoaccepting tRNAs. Dendrogram, dipicting relations between tRNA sets with different amino acid specificities, presents the results of analysis. The method allowed to interpret dendrogram, as a reflection of final stage of the evolution of ambiguous molecules-adaptors to highly specific tRNAs. We have proposed a model of amino acid codons assimilation at this stage. According to the model, the final formation of specific adaptors was followed by assimilation of codon clusters (four codons, differing in the third position), the first dinucleotides of which were connected by transitions. Alternate steps along double-stranded coding DNA accompanied the establishment of tRNA sets with strict specificities. Complementary codons, to which specific adaptors were assigned at step the last, were involved into the process afterwards. PMID- 9390282 TI - Using Tcl for molecular visualization and analysis. AB - Reading and manipulating molecular structure data is a standard task in every molecular visualization and analysis program, but is rarely available in a form readily accessible to the user. Instead, the development of new methods for analysis, display, and interaction is often achieved by writing a new program, rather than building on pre-existing software. We present the Tcl-based script language used in our molecular modeling program, VMD, and show how it can access information about the molecular structure, perform analysis, and graphically display and animate the results. The commands are available to the user and make VMD a useful environment for studying biomolecules. PMID- 9390283 TI - On some operations suggested by genome evolution. AB - Three operations involved in the genome evolution namely, inversion, transposition and duplication, are considered as operations on strings and languages. We show that, for any pair of these operations, there is a language family which is closed under one of the operations and not closed under the second one, however, under some mild conditions the closure of a language family under one of the operations implies that it also closed with respect to another one. PMID- 9390284 TI - The inverse protein folding problem: self consistent mean field optimisation of a structure specific mutation matrix. AB - The goal of the inverse folding problem is to supply a list of sequences compatible with a known protein structure. If two-body interactions are taken into account in energy calculations, an exhaustive exploration of the energy landscape in sequence space cannot be achieved because of the huge number of possible combinations. To circumvent this problem, we propose a method in which multiple copies corresponding to every possible side-chain type are attached to each C alpha position in the protein. The weights of each copy (stored in the sequence matrix SM) are refined using mean field theory: each side-chain copy interacts with the mean field generated by all possible side-chain copies at neighbouring positions, weighted by their respective probabilities. The potential energy is simply taken to be amino acid pair potentials of mean force. The method converges in a few cycles to a self-consistent solution. The refined matrix does not depend on the starting point; therefore the method succeeds in removing memory effects. Starting solely from the backbone of the known structure, and without information from the initial sequence, the final sequence matrix SM is shown to be able to retrieve significant sequence information, as observed through a series of structure-recognizes-sequence(s) computer experiments. The issue of specificity is discussed in detail. PMID- 9390285 TI - Theoretical and algorithmical optimization of the dead-end elimination theorem. AB - The dead-end elimination theorem has proved to be a powerful method to reduce the theoretically accessible conformational space when modeling protein side chains by using a rotameric representation of possible conformations. In this work, theoretical details about variants to the original criterion are discussed. We also provide information on how the equations can be algorithmically implemented in such a way that both computational performance and structural accuracy are optimized. In addition, we discuss the theoretical and practical aspects of three new methods called the "bottom line theorem", dead-end elimination assisted by local modeling and a combinatorial search combined with conventional dead-end elimination. It is shown that the algorithm in its current from enables the determination of the global minimum energy side chain conformation of large proteins on a time scale of hours while for small proteins of up to 30 residues the calculations are done on a time scale of seconds. The latter opens a way to combine a main chain sampling algorithm with the dead-end elimination method to locally model entire fragments of a protein chain. PMID- 9390286 TI - Using views for retrieving data from extremely heterogeneous databanks. AB - Information in molecular biology or biology in general is contained in a multitude of different databanks each specializing in a certain area. This specialization is useful since it improves the maintainability of the data and the flexibility of the overall structure which until now manages to exist without almost any standards. However, information gathering from these extremely heterogeneous sources is difficult since the desired data may be scattered over many databanks. This paper presents the concept of views implemented in the retrieval system SRS that uses links between databanks and a sophisticated parsing engine for information extraction to provide a flexible way of searching and obtaining data across databank boundaries. The implementation provides homogeneous access to all databanks using two types of views: the list-view which displays selected data-fields in their original format and the table-view available for HTML and plain ASCII text which tries to represent the data in a format independent from that of the source databank. PMID- 9390287 TI - Length scales of lipid dynamics and molecular dynamics. AB - Following a brief overview of length scales and system size in computer simulation, it is demonstrated that a simulation sized lipid bilayer (typically a 50 x 50 A2 patch) is in the regime where stretching dominates undulation, while the reverse holds for a flaccid macroscopic membrane. Then it is estimated that current system sizes of membrane simulations must be increased by at least a factor of 10 before thermodynamic limits are approached for quantities such as surface tension. PMID- 9390288 TI - Specific modelling of regulatory units in DNA sequences. AB - Transcriptional control regions are usually composed of a complex arrangement of individual transcriptional elements like protein binding sites. This modular structure allows generation of enormous functional diversity of regulatory regions with a limited set of individual elements. We implemented simple formal representations of these general features of regulatory regions into an algorithm capable of developing complex models reflecting both the element composition and the functional organization of individual elements. Our method (ModelGenerator) requires a training set of at least 10 sequences containing the regulatory regions to be modelled and a very simple initial model which may consist of just two characteristic transcription factor binding sites. We show the capability of our algorithm to expand the initial model solely by comparative sequence analysis leading to complex, biologically meaningful models. A second program (ModelInspector) is capable to scan new sequence data for matches to models defined by ModelGenerator. We show two models for retroviral transcriptional control regions to be highly specific. A search against GenBank using one of the models is shown to be free of false negatives and to produce less than 2 false positives/million nucleotides. Thus, our algorithms appear to be useful tools for the analysis of extremely long genomic sequences which are now becoming available as results of various genome sequencing projects. PMID- 9390289 TI - Test tube systems with cutting/recombination operations. AB - We introduce test tube systems based on operations that are closely related to the splicing operation, i.e. we consider the operations of cutting a string at a specific site into two pieces with marking them at the cut ends and of recombining two strings with specifically marked endings. Whereas in the splicing of two strings these strings are cut at specific sites and the cut pieces are recombined immediately in a crosswise way, in CR(cutting/recombination)-schemes cutting can happen independently from recombining the cut pieces. Test tube systems based on these operations of cutting and recombination turn out to have maximal generative power even if only very restricted types of input filters for the test tubes are used for the redistribution of the contents of the test tubes after a period of cuttings and recombinations in the test tubes. PMID- 9390290 TI - Organizing and computing metabolic pathway data in terms of binary relations. AB - A new database system named KEGG is being organised to computerize functional aspects of genes and genomes in terms of the binary relations of interacting molecules or genes. We are currently working on the metabolic pathway database that is composed of three interconnected sections: genes, molecules, and pathways, which are also linked to a number of existing databases through our DBGET retrieval system. Here we present the basic concept of binary relations and hierarchical classifications to represent the metabolic pathway data. The database operations are then defined as an extension of the relational operations, and the path computation problem is considered as a deduction from binary relations. An example of using KEGG for the functional prediction of genomic sequences is presented. PMID- 9390291 TI - Development of a cell signaling networks database. AB - In multicellular organisms cell signaling networks play important roles in wide range of biological phenomena, such as development, differentiation, reproduction, morphogenesis, carcinogenesis, apoptosis, and even learning. In order to explain biological phenomena based on cell signaling models, we have developed a database for cell signaling networks. The system contains mechanisms of signal transduction and structure and functional data and references of extracellular chemicals and biomolecules. CSNDB is constructed using ACEDB system, and includes various graphical representations such as pathway diagrams, map diagrams, 3-D images, pictures, and VRML environment. The system will be useful for modeling cells and their information processing, and to explain important biological phenomena based on these models. PMID- 9390292 TI - The native sequence determines sidechain packing in a protein, but does optimal sidechain packing determine the native sequence? AB - Globular proteins have highly compact structures and the corresponding packing interactions are widely considered as the principal determinant of the native structure. It is therefore important that theoretical approaches to protein design explicitly take in account packing, which requires that a full atomic representation of the designed protein is maintained. As a first step towards this goal, we have developed in this report an inverse folding algorithm with the aim of specifically designing amino acid sequences which optimise sidechain packing for a given protein fold. The design is performed by a global Monte Carlo optimisation in sequence space, with constant amino acid composition and a full atom representation of the various protein models. Packing is defined by a Lennard-Jones potential. The program was tested by designing stable sequence variants for the chymotrypsin inhibitor fold. The final protein models showed an increase in intramolecular atomic contacts and a decrease in the overall volume compared to the native structure. Starting from the backbone only of the target structure, the algorithm did gradually retrieve reliable though limited sequence information. Higher compatibility might be achieved by improving the potential, however our results suggest that packing interactions are an essential element of a yet-to-be-defined successful energy function for protein design. PMID- 9390293 TI - Design of hydrophobic core of E. coli malate dehydrogenase based on the side chain packing. AB - We have developed computational programs for the de novo design of hydrophobic cores of proteins. The first program optimizes side-chain conformations using an updated rotamer library for potential hydrophobic residues, based on the backbone structure of the protein of interest. The second program selects candidates to be engineered among the sequences by estimating changes in Gibbs free energy between the folded and unfolded structure of the proteins with new sequence. Using these programs, we constructed several variants of E. coli malate dehydrogenase (eMDH) which could have increased stability at 25 degrees C, compared to the wild type enzyme. To quantitate stability change between variants and the wild type, circular dichroism spectra were measured as a function of guanidine hydrochloride concentration at 25 degrees C, pH 7.0. This analysis showed that three variants constructed in this study were stabilized more than or equal to the wild type. This demonstrated that our programs may be powerful tools to design new proteins with high stability. PMID- 9390294 TI - Latent periodicity of DNA sequences of many genes. AB - A method of latent periodicity search being developed. Mutual information is used to reveal of DNA or mRNA sequence latent periodicity. The latent periodicity of DNA sequence is a periodicity with low level of homology between any two periods inside DNA sequence. The mutual information between artificial numerical sequence and DNA sequence is calculated. The length of the artificial sequence period is varied from 2 to 150. The high level of the mutual information between artificial and DNA sequences allows to find any type of latent periodicity of DNA sequence. The latent periodicity of many DNA coding regions has been found. Potential significance of latent periodicity is discussed. PMID- 9390295 TI - Integrating database homology in a probabilistic gene structure model. AB - We present an improved stochastic model of genes in DNA, and describe a method for integrating database homology into the probabilistic framework. A generalized hidden Markov model (GHMM) describes the grammar of a legal parse of a DNA sequence. Probabilities are estimated for gene features by using dynamic programming to combine information from multiple sensors. We show how matches to homologous sequences from a database can be integrated into the probability estimation by interpreting the likelihood of a sequence in terms of the bit-cost to encode a sequence given a homology match. We also demonstrate how homology matches in protein databases can be exploited to help identify splice sites. Our experiments show significant improvements in the sensitivity and specificity of gene structure identification when these new features are added to our gene finding system, Genie. Experimental results in tests using a standard set of annotated genes showed that Genie identified 95% of coding nucleotides correctly with a specificity of 91%, and 77% of exons were identified exactly. PMID- 9390296 TI - Packing as a structural basis of protein stability: understanding mutant properties from wildtype structure. AB - Modeling of protein core mutations using sidechain packing can forecast their effects on stability. We have assessed the structural basis of this approach, by evaluating the accuracy of our 1991 model of a three-site mutant of lambda repressor (V36L/M40L/V47I), against the recently reported crystal structure. The three mutated residues matched the crystal structure to within 0.89A (1.11A for sidechain atoms), giving fairly accurate sidechain placement and packing (81-99th percentile rank in coordinate accuracy). However, the model used different sidechain torsional angles than seen in the crystal structure at residues 36 and 40, apparently to compensate for the backbone shifts present in the actual mutant structure, but not accounted for in our modeling method. To understand the structural basis of stability across a set of lambda repressor core mutants, we have analyzed the mutant models, revealing several simple packing effects: V36I, predicted to be stabilized by filling a hydrophobic cavity; M40V, destabilized by a steric clash with the unusual structural demands of a helix-turn transition. These effects illustrate how mutant stability can often be understood directly from scrutiny of wildtype structure. Simply adding the calculated energies of neighboring point mutations predicts the stability effect of the combined mutant relatively well, with little apparent cooperativity, yielding simple rules for each site's amino acid preferences. Our treatment of core packing indicates that it can permit a large fraction of sequences to fit the native fold, as observed experimentally, far more than indicated by rotamer hard-sphere models. PMID- 9390297 TI - Facilities for exploring molecular biology databases on the Web: a comparative study. AB - We discuss criteria for evaluating and comparing the main facilities provided by molecular biology databases (MBDs) for exploring (that is, retrieving and interpreting data) on the Web. We use these criteria for examining the facilities supported by typical MBDs such as Genbank, AtDB, GSDB, GDB, and MGD (as of September 5, 1996). PMID- 9390298 TI - Towards a bacteriorhodopsin-silicon neuromorphic photosensor. AB - We describe our efforts towards constructing a hybrid protein-silicon neuromorphic photosensor based on the photo-active protein bacteriohodopsin. This protein displays an differential photosensitivity similar to the response of the receptive field of an X-type retinal ganglion cell. Similar bacteriohodopsin photoelectrode arrays display inherent edge detection and motion enhancement. We discuss challenges associated with constructing and understanding the protein silicon interface and possible chemical solutions for our experimental device. PMID- 9390299 TI - An approach to detection of protein structural motifs using an encoding scheme of backbone conformations. AB - This paper presents an approach to detection of protein structural motifs. In our approach, first all protein backbone conformations are converted into character strings using an encoding scheme. Then we use the Smith-Waterman local alignment algorithm to detect common structural motifs. By comparing results with the PROSITE regular expression patterns, our method can detect several motifs which the PROSITE patterns fail to detect. PMID- 9390300 TI - An algorithm to assemble pathways from processes. AB - To understand or to modify a biological pathway, the first step is to determine the patterns of coupling among its processes that are compatible with its input output relation. Algorithms for this purpose have been devised for metabolic pathways, in which the reactions typically leave the enzymes unmodified. As shown here, one of these algorithms can also assemble molecular networks in which reactions modify proteins, if the proteins are included among the inputs to the reactions. Thus one procedure suffices to assemble pathways for metabolism, cytoplasmic signal transduction, and gene regulation. PMID- 9390301 TI - Toward a virtual-labo-system for metabolic engineering: development of biochemical engineering system analyzing tool-kit (BEST-KIT). AB - BEST-KIT is an efficient and user-friendly "biochemical engineering system analyzing tool-kit" integrated the following key modules: 1) mathematical modeling and editing of reaction-scheme, 2) automatic derivation of differential equations, 3) numerical calculation, 4) nonlinear optimization, 5) visualization, 6) retrieve the information on reaction mechanism and kinetic parameters from data-base of metabolic pathways. The users of this simulator are assumed to be unfamiliar with computer technology and with computer programming. The integrated interface (UNIX version) is based on Xlib, XToolkit and OSF/Motif Widget. PMID- 9390302 TI - Method for low resolution prediction of small protein tertiary structure. AB - A new method for the de novo prediction of protein structures at low resolution has been developed. Starting from a multiple sequence alignment, protein secondary structure is predicted, and only those topological elements with high reliability are selected. Then, the multiple sequence alignment and the secondary structure prediction are combined to predict side chain contacts. Such contact map prediction is carried out in two stages. First, an analysis of correlated mutations is carried out to identify pairs of topological elements of secondary structure which are in contact. Then, inverse folding is used to select compatible fragments in contact, thereby enriching the number and identity of predicted side chain contacts. The final outcome of the procedure is a set of noisy secondary and tertiary restraints. These are used as a restrained potential in a Monte Carlo simulation of simplified protein models driven by statistical potentials. Low energy structures are then searched for by using simulated annealing techniques. Implementation of the restraints is carried out so as to take into account of their low resolution. Using this procedure, it has been possible to predict de novo the structure of three very different protein topologies: an alpha/beta protein, the bovine pancreatic trypsin inhibitor (6pti), an alpha-helical protein, calbindin (3icb), and an all beta- protein, the SH3 domain of spectrin (1shg). In all cases, low resolution folds have been obtained with a root mean square deviation (RMSD) of 4.5-5.5 A with respect to the native structure. Some misfolded topologies appear in the simulations, but it is possible to select the native one on energetic grounds. Thus, it is demonstrated that the methodology is general for all protein motifs. Work is in progress in order to test the methodology on a larger set of protein structures. PMID- 9390303 TI - Multiple model approach--dealing with alignment ambiguities in protein modeling. AB - Sequence alignments for distantly homologous proteins are often ambiguous, which creates a weak link in structure prediction by homology. We address this problem by using several plausible alignments in a modeling procedure, obtaining many models of the target. All are subsequently evaluated by a threading algorithm. It is shown that this approach can identify best alignments and produce reasonable models, whose quality is now limited only by the extent of the structural similarity between the known and predicted protein. Using a similar approach structure prediction for the oxidized dimer of S100A1 protein, for which the structure is not known, is presented. PMID- 9390304 TI - Search for DNA conformational features for functional sites. Investigation of the TATA box. AB - A method for search of DNA conformational features significant for functional sites is developed. The method uses helical angles averaged for known X-ray structures. Nucleotide sequences are assigned mean angles in a given region. Choice of the significant angles is based on their capabilities to discriminate functional sites from random sequences. The yeast, invertebrate and vertebrate TATA boxes are analyzed using this method. Regions neighboring the TATA boxes are found to have smaller helical twist and roll angles. The results agree with the experimental data on Dickerson-Drew dodecamers. There is a significant decrease in the length of a small roll angle region with increasing complexity of taxon organization. PMID- 9390306 TI - Exploring the fitness landscapes of lattice proteins. AB - We present methods to investigate the sequence to structure relation for proteins. We use random structures of HP-type lattice models as a coarse grained model to study generic properties of biopolymers. To circumvent the computational limitations imposed by most lattice protein folding algorithms we apply a simple and fast deterministic approximation algorithm with a tunable accuracy. We investigate ensemble properties such as the conditional probability to find structures with a certain similarity at a given distance of the underlying sequence for various alphabets. Our results suggest that the structure landscapes for lattice proteins are generally very rugged, while larger alphabets fine tune the folding process and smoothen the map. This implies a simplification for evolutionary strategies. The applied methods appear to be helpful in the study of the complex interplay between folding strategies, energy functions and alphabets. Possible implications to the investigation of evolutionary strategies or the optimization of biopolymers are discussed. PMID- 9390305 TI - Algorithmic complexity of growth hormone release in humans. AB - Most hormones are secreted in an pulsatile rather than in a constant manner. This temporal pattern of pulsatile hormone release plays an important role in the regulation of cellular function and structure. In healthy humans growth hormone (GH) secretion is characterized by distinct pulses whereas patients bearing a GH producing tumor accompanied with excessive secretion (acromegaly) exhibit a highly irregular pattern of GH release. It has been hypothesized that this highly disorderly pattern of GH release in acromegaly arises from random events in the GH-producing tumor under decreased normal control of GH secretion. Using a context-free grammar complexity measure (algorithmic complexity) in conjunction with random surrogate data sets we demonstrate that the temporal pattern of GH release in acromegaly is not significantly different from a variety of stochastic processes. In contrast, normal subjects clearly exhibit deterministic structure in their temporal patterns of GH secretion. Our results support the hypothesis that GH release in acromegaly is due to random events in the GH-producing tumorous cells which might become independent from hypothalamic regulation. PMID- 9390307 TI - Accurate mean-force pairwise-residue potentials for discrimination of protein folds. AB - We present two new sets of energy functions for protein structure recognition. The first set of potentials is based on the positions of alpha- and the second on positions of beta-carbon atoms of amino acid residues. The potentials are derived using a theory of Boltzmann-like statistics of protein structure by Finkelstein et al. The energy terms incorporate both long-range interactions between residues remote along a chain and short-range interactions between near neighbors. Distance-dependence is approximated by a piecewise constant function defined on intervals of equal size. The size of this interval is optimized. A database of 222 non-homologous proteins was used both for the derivation of the potentials, and for the "threading" test originally suggested by Hendlich et al. For threading, we used 102 non-homologous protein chains of 60 to 200 residues. The energy of each of the native structures was compared with the energy of 45 to 20 thousand alternative structures generated by threading. Of these 102 native structures 94 have the lowest energy with alpha-carbon-based potentials, and even more, 100 of these 102 structures, have the lowest energy with the beta-carbon based potentials. PMID- 9390308 TI - Real time surface reconstruction for moving molecular fragments. AB - Recently we introduced the Reduced Surface as an efficient tool to built molecular surfaces. We describe here how this geometric construct can be used to efficiently reconstruct the solvent excluded surface of a protein for which the coordinates of a subset of atoms are changing. We show that, the complexity of that operation is not dependent upon the size of the molecule and is in O[tlog(t)] where t is the maximum of the number of probes and atoms involved in the reconstruction of the surface. The algorithms described here have been implemented and tested on several proteins. The triangulation of the solvent excluded surface of proteins in which a side chain was changing conformation could be updated at rates ranging from 7 to 22 frames per second. We also applied this method to compute the surface area fluctuation of the FIV protease undergoing a constrained molecular dynamics simulation (16 mobile residues). Rate of 6 frames per second were obtained in this case. PMID- 9390309 TI - Finding association rules on heterogeneous genome data. AB - A novel approach for discovery of knowledge from genome data, which has been recently watched with interest in the research area of database, is applied to finding unified rules spreading over sequence, structure, and function of protein. As the result of experiments using data extracted from PDB, SWISS-PROT, and PROSITE, some association rules stating sequential/structural/functional aspects of two kinds of endopeptidases were found. PMID- 9390310 TI - Enumerating suboptimal alignments of multiple biological sequences efficiently. AB - The multiple sequence alignment problem is very applicable and important in various fields in molecular biology. Because the optimal alignment that maximizes the score is not always biologically most significant, providing many suboptimal alignments as alternatives for the optimal one is very useful. As for the alignment of two sequences, this suboptimal problem is well-studied, but for the alignment of multiple sequences, it has been considered impossible to investigate such suboptimal alignments because of the enormous size of the problem. The optimal multiple alignment can be obtained with A* algorithm, and an efficient algorithm for the k shortest paths problem on general graphs is discovered recently. We extend these algorithms for computation of set of all aligned groups of residues in optimal and suboptimal alignments, and for enumeration of suboptimal alignments. The suboptimal alignments are numerous. Thus we discuss what kind of suboptimal alignment is unnecessary to enumerate, and propose an efficient technique to enumerate only necessary alignments. The practicality of these algorithms are demonstrated through experiments. Moreover, the property of suboptimal alignments of multiple sequences are also examined through experiments. PMID- 9390311 TI - Redox properties of cytochrome c: novel linear response and hybrid continuum microscopic methodologies. AB - Redox properties of yeast cytochrome c are estimated using molecular dynamics combined with a simple linear response approximation, as well as a hybrid continuum-molecular dynamics (COMD) approach. In both approaches, the free energy associated with an electrostatic perturbation (a redox electron) is separated into its relaxation and static (non-relaxation) components. The static component is calculated from the molecular dynamics simulation. The relaxation component is then calculated with a linear response approximation, either from the molecular dynamics, or from a separate continuum calculation. This latter hybrid approach exploits the relative robustness of continuum models for dealing with large perturbations, while avoiding some of their limitations. It is quite general, and could be applied for example to pKa calculations. PMID- 9390312 TI - Protein superfamily members as targets for computer modeling: the carbohydrate recognition domain of a macrophage lectin. AB - Members of protein superfamilies display similar folds, but share only limited sequence identity, often 25% or less. Thus, it is not straightforward to apply standard homology modeling methods to construct reliable three-dimensional models of such proteins. A three-dimensional model of the carbohydrate recognition domain of the rat macrophage lectin, a member of the calcium-dependent (C-type) lectin superfamily, has been generated to illustrate how information provided by comparison of X-ray structures and sequence-structure alignments can aid in comparative modeling when primary sequence similarities are low. PMID- 9390313 TI - Definite-clause grammars for the analysis of cis-regulatory regions in E. coli. AB - Based on an extensive collection of sigma 70 associated regulatory mechanisms, a grammatical model has been constructed that define the functional positions and combinations of sites within DNA regulatory regions. The syntactic rules and the dictionary implemented in a Prolog program were coupled to consensus matrices used as "sensors" to integrate a syntactic recognizer. A systematic comparison between the syntactic recognizer and the standard weight matrix methodology is presented using 12 regulatory proteins and the whole collection of about 130 sigma 70 DNA regulatory regions. On the average an increased sensitivity of 5 to 10 fold is obtained with this novel approach. PMID- 9390314 TI - Enumeration of flux routes through complex biochemical reactions. AB - In the present work, a general algorithm for enumeration of the flux routes via which a chemical species "flows" through a complex biochemical reaction is outlined and presented by way of a biological example, a kinetic model for potassium ion permeation through a voltage-gated ion channel. The algorithm is readily amenable to computer based implementation and when used in conjunction with an existing algorithm provides a convenient means for simulation of the equilibrium and steady-state isotope exchange kinetics of complex biochemical reactions. PMID- 9390315 TI - Using the radial distributions of physical features to compare amino acid environments and align amino acid sequences. AB - We have performed a comprehensive analysis of the microenvironments surrounding the twenty amino acids. Our analysis includes comparison of amino acid environments with random control environments as well as with each of the other amino acid environments. We describe the amino acid environments with a set of 21 features summarizing atomic, chemical group, residue, and secondary structural features. The environments are divided into radial shells of 1 A thickness to represent the distance of the features from the amino acid C beta atoms. We make the results of our analysis available graphically over the world wide web. To illustrate the validity and utility of our analysis, we used the amino acid comparative profiles to construct a substitution matrix, the WAC matrix, based on a simple summary of the computed environmental differences. We compared our matrix to BLOSUM62 and PAM250 in BLAST searches with query sequences selected from 39 protein families found in the PROSITE database. Although BLOSUM62 was the most sensitive matrix overall, our matrix was more sensitive for some families, and exhibited overall performance similar to PAM250. Our results suggest that the radial distribution of biochemical and biophysical features is useful for comparing amino acid environments, and that similarity matrices based on the geometric distribution of features around amino acids may produce improved search sensitivity. PMID- 9390316 TI - TRANSFAC database as a bridge between sequence data libraries and biological function. AB - The TRANSFAC database contains information about regulatory DNA sequences and the proteins (transcription factors) binding to and acting through them. It may thus serve as a dictionary for the biological meaning of these sequence elements. Moreover, the TRANSFAC data can be used to describe these elements, to define consensi and matrices for elements of certain function, and thus to provide means of identifying regulatory signals in newly unravelled genomic sequences. PMID- 9390317 TI - A new approach to protein fold recognition based on Delaunay tessellation of protein structure. AB - We propose new algorithms for sequence-structure compatibility (fold recognition) searches in multi-dimensional sequence-structure space. Individual amino acid residues in protein structures are represented by their C alpha atoms; thus each protein is described as a collection of points in three-dimensional space. Delaunay tessellation of a protein generates an aggregate of space-filling, irregular tetrahedra, or Delaunay simplices. Statistical analysis of quadruplet residue compositions of all Delaunay simplices in a representative dataset of protein structures leads to a novel four body contact residue potential expressed as log likelihood factor q. The q factors are calculated for native 20 letter amino acid alphabet and several reduced alphabets. Two sequence-structure compatibility functions are computed as (i) the sum of q factors for all Delaunay simplices in a given protein, or (ii) 3D-1D Delaunay tessellation profiles where the individual residue profile value is calculated as the sum of q factors for all simplices that share this vertex residue. Both threading functions have been implemented in structure-recognizes-sequence and sequence-recognizes-structure protocols for protein fold recognition. We find that both profile and total score based threading functions can distinguish both the native fold from incorrect folds for a sequence, and the native sequence from non-native sequences for a fold. PMID- 9390318 TI - Dental care associated with an outbreak of HIV infection among dialysis patients. AB - An outbreak of 14 cases of human immunodeficiency virus (HIV) infection was discovered by chance in May 1993 among hemodialysis patients at a university hospital in Bucaramanga, Colombia. The outbreak occurred in 1992. Stored sera were used to establish the probable period of infection (PPI) for 10 of the 14 cases. A nested case-control study was carried out to evaluate possible transmission mechanisms. The health care experience of each HIV-positive patient during that patient's PPI was compared to the experience of time-matched controls. Only invasive dental procedures were significantly associated with the risk of infection. Patients upon whom invasive dental procedures were performed during their PPIs had an average risk of HIV infection 8.15 times greater than comparable controls (P = 0.006), and seven out of nine cases of HIV infection with known PPIs in 1992 had an invasive dental procedure performed one to six months before seroconversion. None of the dental care personnel were found to be infected. Based on the available evidence, it seems most likely that the infection was transmitted from patient to patient by contaminated dental instruments. PMID- 9390319 TI - Virtual elimination of iodine deficiency disorders in Bolivia. PMID- 9390320 TI - [Nefazodon (Nefadar)--a new dual serotoninergic antidepressant. Introductory press conference: "Nefazodone--new perspectives in antidepressive therapy." 25-26 April 1997]. PMID- 9390321 TI - The inherited palmoplantar keratodermas. AB - The inherited palmoplantar keratodermas (PPK) constitute a complex heterogeneous group of genodermatoses, which are difficult to classify clinically. The application of modern molecular biology techniques are leading to an increased understanding of the genetic bases of these disorders and are paving the way towards a classification based upon molecular pathology. We review the recent research advances in this field and the implications for development of novel approaches to disease management. PMID- 9390322 TI - Topical FK506: a potent immunotherapy for alopecia areata? Studies using the Dundee experimental bald rat model. AB - We elected to examine the efficacy of the topically applied immunosuppressive agent FK506 (Prograf) in the treatment of alopecia areata (AA) using the Dundee experimental bald rat (DEBR) model. Thirty lesional DEBR rats were allocated to five groups of six. Group 1 rats received 0.1 mL of a 0.25% solution of FK506 within a 2 x 2 cm marked area on one bald flank twice a week (125 micrograms FK506/cm2 per week) for 8 weeks, while the contralateral flank was left untreated. In group II, 0.05 mL of a 0.1% solution of FK 506 was applied 5 days per week on one flank (62.5 micrograms FK506/ cm2 per week) and control vehicle to the opposite flank for 8 weeks. Group III rats were treated as in group II except that drug and vehicle were applied twice a week (25 micrograms FK506/cm2 per week) for 4 weeks. A positive control group received orally administered cyclosporin A (CsA) (10 mg/kg daily) for 8 weeks and a further group was left untreated. Rats were regularly examined and photographed with skin biopsies taken from groups II and III. All FK 506-treated rats regrew hair at the site of drug application within 14-21 days. Growth continued for 3 weeks beyond termination of treatment after which gradual hair loss was observed. No hair growth was seen as a result of vehicle application and hair loss continued on untreated areas and in the untreated control group. Immunohistology revealed a drastic reduction in the follicular inflammatory infiltrate at the site of the FK506 application. The oral CsA group responded by simultaneous regrowth of hair over the whole body. Our findings suggest that FK506 may have considerable potential as a topical treatment for AA. PMID- 9390323 TI - Serum-free culture of wool follicles: effects of nutrients, growth factors and hormones. AB - A serum-free culture system allowed the continued growth of fibre from follicles for 8-10 days. Fibre growth was responsive to changes in the level of calcium, glucose and amino acids in the culture medium, and was stimulated by the inclusion of insulin (10 micrograms/mL) in the medium. Culture of follicles in the presence of conditioned media from dermal papilla cells or of mitomycin treated dermal papilla cells had no effect on fibre growth. Neither thyroid hormones nor hydrocortisone altered fibre growth. The progressive decline in fibre growth during follicle culture was accompanied by morphological changes in the follicle bulb. Oxidative damage did not appear to be the cause of these changes as there was no increase in fibre growth rate or longevity when antioxidants were used. This model provides a useful system to study the direct effects of various hormonal, nutritional and growth factors of fibre growth and follicle metabolism. PMID- 9390324 TI - Postsurgical wound progression monitored by temporal changes in the expression of matrix metalloproteinase-9. AB - It is important to monitor the early stages of postoperative wound repair in order to identify those problems associated with impaired healing. Many of the crucial cellular responses of early wound healing, such as inflammatory infiltration, angiogenesis and re-epithelialization, are made possible through the action of matrix metalloproteinases (MMPs). Expression of MMP-2 and MMP-9 is elevated in acute wounds, and still greater levels are found in chronic wounds, indicating that uncontrolled proteolysis is a characteristic of retarded healing. Therefore, comparative measurements of MMPs may be used to monitor the progression of early wound healing. To investigate this, wound fluids and sera were collected from mastectomy and colectomy patients throughout early stages of repair, and the temporal expression profile established. Wounds which were healing were expressed maximal levels of MMP-9 at 24 h, followed by a significant decline by 48 h. Persistent elevation of MMP-9 expression was associated with infected and chronic wounds, and was identified in postoperative wounds by the absence of the significant decline between 24 and 48 h. Measurement of MMP-9 in postoperative wound fluids, therefore, provides an early indicator of impaired healing, which may be evaluated non-invasively within 48 h of closure. PMID- 9390325 TI - Increased expression of gelatinases A and B by skin explants from patients with anetoderma. AB - The extent of alterations to the elastic fibre network in lesional skin areas of three patients with anetoderma was assessed by quantitative image analysis of tissue sections and compared with morphometric parameters from unaffected sites of the same individuals. In the anetodermic skins pre-elastic fibres were undetectable or extremely rare: the volume fraction (Vv%) occupied by these pre elastic fibres was 0-0.3%, while in unaffected skins the Vv% occupied by pre elastic fibres was 0.5-0.8%. A nearly complete absence of dermal elastic fibres in lesional skins from the three patients was evidenced (Vv% = 0.2-0.3%). Organ cultures were performed using explants from skin with or without anetodermic lesions to quantify the expressions of elastase-type proteinases. All tissues from anetodermic lesions expressed proforms of gelatinases A and B and the activated form of gelatinase A; their levels increased with the culture time. In comparison, enzymatic activities on oligopeptide substrates specific for leucocyte elastase and fibroblast plasma membrane-associated metalloelastase were not detected in the conditioned media of any explants at any time of culture from 1 to 5 days. Increased production of progelatinases A and B and activation of progelatinase A could be mainly responsible for the degradation of skin elastic fibres demonstrated in anetodermic skins. PMID- 9390326 TI - Linear IgA disease: a report of two dermal binding sera which recognize a pepsin sensitive epitope (?NC-1 domain) of collagen type VII. AB - Linear IgA disease is a subepidermal blistering disease characterized by IgA autoantibody deposition at the basement membrane zone of skin and mucosa. The antigens targeted in linear IgA disease have been defined by their molecular weight and localization. It has been proposed that a minority of linear IgA disease sera that bind to the dermal aspect of salt-split skin target collagen type VII. We have identified two patients with linear IgA disease using dermal binding IgA autoantibodies on salt-split skin which recognize collagen type VII by immunoblot analysis. The reactive epitope was destroyed by the proteolytic enzymes pepsin, which destroys the NC-1 domain of collagen type VII, and protease type VIII. Localization studies compared the IgA autoantibodies from these patients with the monoclonal antibody LH7.2 to the NC-1 domain of collagen type VII and showed colocalization on a dermal cylindroma tumour tissue, and a similar distribution with immunogold electron microscopy, using purified blister fluid from one patient. We propose from these results that the IgA autoantibodies from these two patients recognize the NC-1 domain of collagen type VII, the classical immunodominant epitope for epidermolysis bullosa acquisita. PMID- 9390327 TI - One single erythemagenic UV irradiation is more effective in increasing the proliferative activity of melanocytes in melanocytic naevi compared with fractionally applied high doses. AB - The effect of a single irradiation with UV light on the expression of Ki67 antigen, topoisomerase II alpha, proliferating cell nuclear antigen (PCNA), the melanocyte activation marker HMB-45 and protein p53 in melanocytic naevi was investigated 1 week after application of a single erythemagenic UV dose and after daily exposures with suberythemagenic doses over 4-6 weeks. To assess the effect of UV irradiation, one half of each naevus was shielded with black tape during the UV exposure, and the irradiated part and the non-irradiated parts were evaluated separately. Except for HMB-45, a double staining procedure was performed to distinguish between labelled melanocytes and keratinocytes. After semiquantitative assessment of the staining signal the irradiated part was compared with the non-irradiated part of the same naevus. Morphological changes and an enhanced proliferative/ reparative activity in melanocytes were much more frequent in the naevi irradiated with a single erythemagenic UV dose than in those given repeated suberythemagenic doses. In addition, the keratinocytes showed an increased labelling for PCNA and p53 after the single irradiation. These data may support the importance of intermittent UV exposure and sunburns in the development of both benign and malignant melanocytic lesions. PMID- 9390328 TI - Contact allergens and sodium lauryl sulphate upregulate vascular endothelial growth factor in normal keratinocytes. AB - In allergic and irritant contact dermatitis, keratinocytes are major target cells that can be activated to take part in local reactions by secreting soluble mediators. Among the growth factors produced by keratinocytes, vascular endothelial growth factor (VEGF) is a powerful inducer of permeability of endothelial cells, and is involved in inflammation. We determined whether different contact allergens, dinitrosulphobenzene (DNSB), para-phenylenediamine (pPD) and the metals nickel and chromium, as distinct from cobalt, which has been shown to mimic the effects of hypoxia, can modify the basal level of VEGF in normal human keratinocytes when tested at various, non-toxic concentrations. The effects of an irritant, sodium lauryl sulphate (SLS), and of hydrocortisone were also tested. Our results showed an intense dose-dependent upregulation of VEGF release by keratinocytes after treatments by metals, pPD and SLS. DNSB induced only a moderate increase of VEGF. Hydrocortisone reduced the basal level as well as the nickel-induced upregulation of VEGF. These findings suggest that contact allergens and irritants probably upregulate VEGF in keratinocytes by different mechanisms and may contribute directly to the microvascular hyperpermeability which characterizes both contact and irritant dermatitis. PMID- 9390329 TI - Lack of effect of growth hormone therapy on the count and density of melanocytic naevi in children. AB - An observation of accelerated growth of acquired melanocytic naevi (AMN) during treatment with human growth hormone (GH) raised concerns about the potential risk of melanoma in treated patients. An increased number of AMN, rather than growth rate, is associated with a higher risk for melanoma. It is unknown whether treatment with GH causes an increase in numbers of AMN. We evaluated the effect of GH treatment on the number of AMN in a cross-sectional study of 90 children with GH deficiency. AMN counts and densities in these children were compared with those found in a control group of 100 children. Factors potentially related to increased numbers of AMN, such as age, sex, skin colour, number of episodes of sunburn and duration of GH therapy were determined. Among the various factors, only the age and colour of unexposed skin area were predictive for the total number and density of AMN. No correlation was found between the AMN counts or density and the duration of GH therapy. There was no difference in AMN counts or density between the GH-deficient patient group and the control groups. We conclude that GH therapy in children is not associated with increased AMN count and density and is unlikely to potentiate the risk for melanoma in these children. PMID- 9390330 TI - Extracorporeal photochemotherapy induces apoptosis of infiltrating lymphoid cells in patients with mycosis fungoides in early stages. A quantitative histological study. AB - Extracorporeal photochemotherapy (ExP) is a well-tolerated new form of chemoimmunotherapy, which is considered to be effective for cutaneous T-cell lymphoma (CTCL) and the treatment of choice for Sezary syndrome. Improvements have also been seen in patients with non-erythrodermic mycosis fungoides (MF) in the early stages, even when tumour cells are not detectable in the peripheral blood. In this study, we used ExP as a monotherapy in seven patients who had early stage (Ib) MF, and who were no longer responsive to or had contraindications for other therapies. We observed a clinical improvement in the disease after 12 months of treatment: one patient showed a complete response, five a partial response, and one remained stable. In each patient we compared skin biopsies of large plaque lesions before and after the treatment. We undertook a histological evaluation of the infiltrate. The lymphoid cell proliferation and death rates were quantified using the following parameters; lymphoid cell density (LCD), Ki67 + lymphoid cell nuclei percentage (Ki67 + Lcn percentage), and apoptotic index (AI). Significant decreases in the lymphoid cell infiltrate and in cell proliferation, and a significant increase in AI were observed after therapy. The mean LCD decreased from 187 +/- 33 to 34 +/- 17.7, Ki67 + Lcn mean percentage decreased from 16.9 +/- 3.9 to 4.9 +/- 2.4, and the AI mean value increased from 0.05 +/- 0.03 to 2.41 +/- 1.54. Our results suggest a role for apoptosis in the improvement of the skin lesions and are in line with some reports on the mode of action of ExP. Although the way in which ExP works needs to be clarified further, it does seem to stimulate a CD8+ cell-mediated anticlonotypic activity against circulating pathogenic clones. Furthermore, a release of tumour necrosis factor alpha (TNF-alpha) by circulating monocytes has been demonstrated after ExP. Both are known to induce cell death by apoptosis. PMID- 9390331 TI - Experiences with the severity scoring of atopic dermatitis in a population of German pre-school children. AB - Severity scoring of atopic dermatitis (SCORAD) was introduced as a standard tool but has not been used in a population-based epidemiological study; the objective of the present study was to determine the practicability of this instrument in this setting. We assessed the distribution of the severity of atopic eczema in the community and investigated differences between east and west Germany. A factor analysis was then carried out to characterize the variables of this scoring system and to analyse possible relationships within them. A multicentre cross-sectional study was carried out in five east German and two west German locations in 1994; pre-school children (5-6 years old) were investigated and cases of atopic eczema identified by a dermatological examination. The SCORAD was used to determine the severity of atopic eczema and the results assessed using analysis of variance and principal component analysis (varimax rotation). In all, 1511 (76.2%) of the children originally contacted participated and 11.3% were diagnosed with atopic eczema at the time of examination. The median severity scores was 21.4 (interquartile range 13.5) and there was a tendency to higher scores in west Germany for the mean overall score, the intensity score and the extent. 'Erythema' (1.30 vs. 1.06; P = 0.006) and 'excoriation' (0.77 vs. 0.36; P = 0.002) were significantly more prominent in children with eczema from west Germany (adjusted for observer). Interobserver variabilities of the SCORAD parameters were calculated, adjusted for location and were in accordance with earlier findings. Principal component analysis identified three independent factors accounting for 54.1% of the total variance. A severity factor, characterized by 'extent', 'lichenification', 'excoriation' and 'pruritus', was separated from a factor with an acute eczema-type profile ('erythema', 'oedema', 'oozing') and a factor whose major characteristics were 'extent', 'dryness', and 'sleep loss'. We conclude that atopic eczema is frequent in pre-school children. The SCORAD proved to be readily applicable and useful in epidemiological studies, but further validation is needed. PMID- 9390332 TI - The effectiveness of acne treatment: an assessment by patients of the outcome of therapy. AB - The impact of acne on quality of life can be profound. Although treatment improves the clinical features of acne, there is little information on its benefit from the patients' point of view. In this study, patients with acne referred to a dermatology clinic were sent questionnaires before being seen, and 4 and 12 months afterwards. Clinical severity was assessed by a dermatologist at baseline and at 4 months. Quality of life was assessed by patients using the Short Form 36 instrument (SF-36), the Dermatology Life Quality Index (DLQI), Rosenberg's measure of self-esteem and the General Health Questionnaire (GHQ-28). Of 90 available patients, 79 (89%) returned at least one follow-up questionnaire. The clinical acne grade improved substantially with treatment. There were also significant improvements either at 4 or 12 months in the DLQI, self-esteem. GHQ 28 and all five dimensions of the SF-36 that were impaired at baseline. Quality of life continued to improve between the 4- and 12-month follow-up questionnaires. Clinical and patient-assessed outcomes were significantly better in patients treated with isotretinoin. The study showed that disability caused by acne can be largely reversed by effective treatment. It also showed that patient assessed measures of outcome can respond to changes over time and discriminate between treatments differing in effectiveness. PMID- 9390333 TI - A novel anti-inflammatory drug, SDZ ASM 981, for the topical and oral treatment of skin diseases: in vivo pharmacology. AB - There is a need for safe and effective therapies for inflammatory skin diseases. Current topical and systemic treatment of psoriasis is effective but suffers from side-effects or is inconvenient. The therapeutic armamentarium for atopic dermatitis is very limited and far from satisfactory. In vivo preclinical data are presented for SDZ ASM 981, a novel ascomycin macrolactam derivative with high anti-inflammatory activity. Anti-inflammatory activity was observed in mouse, rat and pig models of allergic contact dermatitis. In the pig model, topical SDZ ASM 981 was as effective as the ultrapotent corticosteroid clobetasol-17-propionate, and when compared with a series of commercial topical corticosteroid preparations, 0.1% SDZ ASM 981 had equivalent efficacy to clobetasol-17 propionate (0.05%), the most potent product on the market. Unlike the corticosteroid, however, SDZ ASM 981 did not cause skin atrophy in pigs. SDZ ASM 981 potently inhibited allergic contact dermatitis in mice and rats when given systemically, and oral treatment was more effective than cyclosporin A in rats. Furthermore, SDZ ASM 981 has a low potential for affecting systemic immune responses, as demonstrated in rat models of localized graft vs. host reaction and allogeneic kidney transplantation. Preclinical results suggest that SDZ ASM 981 has the potential to be a well-tolerated and effective drug for topical as well as oral treatment of inflammatory skin diseases. PMID- 9390334 TI - Toenail onychomycosis in patients with acquired immune deficiency syndrome: treatment with terbinafine. AB - Skin infections caused by dermatophytes are one of the most frequent dermatological complications in patients with acquired immunodeficiency syndrome (AIDS) resulting from infection with human immunodeficiency virus (HIV). Tinea unguium associated with AIDS is characterized by being clinically more aggressive and therapeutically more difficult to treat than in the general population. Terbinafine is considered to be a first-choice option for the treatment of dermatophyte onychomycosis in immunocompetent individuals. This drug has been used in a series of 21 HIV-positive patients diagnosed with tinea unguium for 1 year in the University Hospital La Paz, Madrid. All patients underwent a subsequent clinical follow-up for 6 months. The results showed a high percentage of clinical and mycological cures, as well as maintenance of the response after follow-up; no drug interactions or significant adverse effects related to the drug under study were recorded. PMID- 9390335 TI - Once daily treatment of psoriasis with tacalcitol compared with twice daily treatment with calcipotriol. A double-blind trial. AB - Once daily topical treatment of psoriasis with tacalcitol ointment (4 micrograms/g) was compared with twice daily treatment with calcipotriol ointment (50 micrograms/g) in a double-blind, randomized study over a treatment period of 8 weeks. The severity of pruritus, erythema, infiltration and scaling was scored on a scale from 0 to 4. These features were scored at the initiation of treatment, after 2, 4, 6 and 8 weeks of treatment, and at 4 weeks after discontinuation of treatment. The sum score was the total score for erythema, infiltration and scaling. Serum levels of calcium, phosphate, ionized calcium and intact parathyroid hormone were used as safety parameters. Two hundred and eighty seven adults with stable plaque psoriasis participated and were treated at least once. Both tacalcitol and calcipotriol ointments effectively reduced the severity of psoriasis. The mean reduction in the sum score in the intention-to-treat population of 287 patients was 4.03 in the group treated with tacalcitol compared with 5.05 in the group treated with calcipotriol. The mean baseline sum scores were 7.64 and 7.15, respectively. The acceptability of both ointments was excellent, and none of the patients had adverse effects in terms of increased serum calcium or other alterations in calcium metabolism. Although less effective than calcipotriol ointment used twice daily, tacalcitol ointment is an effective and useful once daily treatment of chronic plaque psoriasis. PMID- 9390336 TI - The combination of oral acitretin and bath PUVA for the treatment of severe psoriasis. AB - We report four patients with severe erythrodermic, pustular psoriasis, or plaque type psoriasis, who were treated with a combination of acitretin and bath PUVA. After 4 weeks out-patient treatment, the psoriasis in all patients had improved by > or = 90%. No patient had relapsed when reviewed at 3 months. No significant side-effects were seen with the combined retinoid/bath PUVA treatment. Acitretin and bath PUVA may be safely combined for the treatment of severe psoriasis. PMID- 9390337 TI - CO2 laser debridement of sulphur mustard (bis-2-chloroethyl sulphide) induced cutaneous lesions accelerates production of a normal epidermis with elimination of cytological atypia. AB - Sulphur mustard (bis-2-chloroethyl sulphide; HD) exposure acutely produces lesions that vary from mild erythema, to blister formation, to necrosis. When blisters occur, with or without necrosis, healing of the lesions is delayed. Weanling pigs exposed to a mild erythema-producing dose of HD and to a moderate erythema-producing dose that consistently gave microblister formation were treated with CO2 laser (Tru-Pulse) debridement at 6, 24 or 48 h after exposure. The histopathological features observed at 14 days after exposure in control skin and skin exposed to both HD doses were compared with the features observed in CO2 laser-debrided skin in non-exposed and HD-exposed skin sites. The overlying epidermis in the non-laser treated lesions was thin, with cytological atypia and squamoid changes within the basal cell layer, as well as scattered apoptotic/necrotic keratinocytes. An increased inflammatory infiltrate and necrobiotic changes in the dermis were seen at the higher HD dose. All laser treated lesions appeared identical, with a thick, differentiated epidermis and a well-formed basal cell layer. There was minimal inflammatory infiltrate. In the papillary dermis there were increased stromal cells. Laser debridement of mild clinical lesions induced by HD produced a more functional epidermis by 14 days as well as clearing the epidermis of damaged keratinocytes. PMID- 9390338 TI - The prevalence of skin disease in HIV infection and its relationship to the degree of immunosuppression. AB - A cross-sectional study of human immunodeficiency virus (HIV) positive patients who attended the HIV clinic in Brighton over a 4-month period was carried out to describe the prevalence and severity of skin manifestations in HIV-positive patients and to elucidate their association with the peripheral CD4 cell count and with the HIV disease stage. The subjects were consecutively examined by an experienced dermatologist. Skin manifestations were classified into infections, dermatoses, pruritus and neoplasm. A severity index was derived by scoring each condition as either absent, mild, moderate or severe. One hundred and fifty-one patients were enrolled with a mean age of 38.3 years. One hundred and thirty-nine were homo/bisexual men; 58 were asymptomatic and 35 had acquired immune deficiency syndrome (AIDS); 37 had CD4 counts below 200. Skin conditions were present in 138 of the 151 subjects (91.4%). The total number of events was 331. The most frequent problem was infection followed by dermatoses, pruritus and malignancy. The most frequent condition was seborrhoeic eczema followed by tinea and xerosis. We have demonstrated a statistically significant association between CD4 count, disease stage and skin manifestations in HIV-positive individuals. PMID- 9390339 TI - A subepidermal blistering disease with histopathological features of dermatitis herpetiformis and immunofluorescence characteristics of bullous pemphigoid: a novel subepidermal blistering disease or a variant of bullous pemphigoid? AB - A 64-year-old man presented with a bullous eruption which clinically and histopathologically resembled dermatitis herpetiformis. However, direct immunofluorescence analysis showed IgG deposits at the basement membrane zone, indicating a relationship with bullous pemphigoid or epidermolysis bullosa acquisita. Indirect immunofluorescence studies on salt-split skin showed binding of IgG mainly on the dermal side of the blister. Immunoblot analysis revealed a novel 200 kDa dermal antigen that could be associated with a major pathogen in this blistering disease. The histopathological similarity to dermatitis herpetiformis and the immunofluorescence findings indicating bullous pemphigoid or epidermolysis bullosa acquisita seem typical of a distinct subepidermal blistering disease characterized by this 200 kDa antigen. However, the pathogenetic role of autoantibodies against this antigen should be further elucidated before confirming whether this case represents a novel subepidermal blistering disease or a special variant of bullous pemphigoid. PMID- 9390340 TI - Human herpesvirus 6 infection associated with anticonvulsant hypersensitivity syndrome and reactive haemophagocytic syndrome. AB - Viral infections are thought to play a part in some cutaneous drug reactions. Human herpesvirus 6 (HHV6), which is the agent of exanthema subitum (sixth disease), has never been implicated in a drug reaction. We report a patient with severe phenobarbital-induced anticonvulsant hypersensitivity syndrome in whom a fulminant haemophagocytic syndrome was associated with HHV6 infection. We discuss the possible role of HHV6 in this reactive condition. PMID- 9390341 TI - Sweet's syndrome and malignancy in the U.K. AB - Acute febrile neutrophilic dermatosis (Sweet's syndrome) is reported to be a marker for underlying malignancy. Much of the evidence for this is based on case reports, small series of cases and reviews of the literature. In order to clarify the association with malignancy and determine the common clinical features of Sweet's syndrome, we reviewed the case notes of patients presenting to six dermatology units in the U.K. Eighty-seven cases of histologically proven Sweet's syndrome were reviewed. Fourteen patients (16%) developed associated malignancy, predominantly haematological, two patients (2%) had a history of previous malignancy and four patients (5%) had premalignant conditions (monoclonal gammopathy, two: myelodysplasia, two). Malignancy developed up to a year after presentation with Sweet's syndrome. Patients with associated malignancy were more likely to be anaemic (P < 0.01) at presentation, had a lower mean platelet count (207 x 10(9)/L vs. 332 x 10(9)/L; P < 0.003) and were, on average, older (59 years vs. 49 years; P = 0.002). Contrary to previous reports, a greater percentage of females developed malignancy than males. PMID- 9390342 TI - Lupus miliaris disseminatus faciei--the DNA of Mycobacterium tuberculosis is not detectable in active lesions by polymerase chain reaction. AB - There has been a controversy as to the origin of lupus miliaris disseminatus faciei (LMDF). It was originally thought to be associated with tuberculosis, due to its histopathological similarity. Recently, this association has been doubted, although there remain reported cases of LMDF associated with Mycobacterium tuberculosis. Three patients with the clinical and histopathological features of LMDF are described. Skin from these patients was analysed by polymerase chain reaction (PCR) using two different oligoprimers for the detection of 123 bp and 165 bp DNA fragments specific for M. tuberculosis complex. With these two PCR systems, no M. tuberculosis DNA was detected in any of the LMDF patients. It was present in all positive controls and absent in all negative controls. In this study we could not demonstrate an association between LMDF and tuberculosis. PMID- 9390343 TI - Aquagenic urticaria and human immunodeficiency virus infection: treatment with stanozolol. AB - We report the first case of aquagenic urticaria in a patient with human immunodeficiency virus (HIV) infection. This is a rare physical urticaria not previously described in this context. The disorder proved unamenable to conventional treatment with antihistamines, but did respond dramatically to stanozolol, suggesting a novel indication for this anabolic steroid. PMID- 9390344 TI - Cutaneous manifestation of Chagas' disease after heart transplantation: successful treatment with allopurinol. AB - We describe two patients who underwent cardiac transplantation for chronic cardiomyopathy of Chagas' disease, and in whom the disease was reactivated with the development of cutaneous lesions. In both cases, the skin lesions regressed completely after 2 months of therapy with allopurinol. PMID- 9390345 TI - Increased plasma interleukin-6 in cutaneous plasmacytoma: the effect of intralesional steroid therapy. AB - Cutaneous plasmacytosis is a rare disorder without systemic plasma cell proliferation in organs other than the skin, with a possible malignant transformation. However, there are few effective therapies available. It has been reported that interleukin-6 (IL-6), which is a cytokine inducing B-cell differentiation to immunoglobulin-producing cells, plays a part in systemic plasmacytosis. In this study, we performed intralesional steroid therapy in the lesions of cutaneous plasmacytosis in three patients, which resulted in sufficient clinical effects. We demonstrated that before treatment, plasma IL-6 levels were significantly elevated in all the patients, and that levels were reduced in parallel with the clinical improvement after therapy. Immunohistochemistry revealed IL-6 protein expression on tumour cells in the lesional skin. Reverse transcription-polymerase chain reaction (RT-PCR) detected IL-6 mRNA in the lesional skin in all cases, levels of which were decreased after the effective intralesional steroid therapy, but which were unchanged after ineffective topical photochemotherapy (PUVA). Peripheral blood mononuclear cells from the patients produced significantly large quantities of IL-6 which were reduced by addition of steroid in vitro. These results suggest that the generation of IL-6 plays the key role in cutaneous plasmacytosis and that intralesional steroid therapy is effective in reducing the production of IL-6 in this disorder. PMID- 9390346 TI - Merkel cell carcinoma arising after therapeutic immunosuppression. AB - Azathioprine and cyclosporin have been used as immunosuppressants for many years, but long-term use has also been associated with neoplasia. We report three cases of rapidly fatal Merkel cell carcinoma in patients who had been treated with azathioprine for many years either for rheumatoid arthritis or following organ transplantation. Two of these patients had also received cyclosporin. We suggest that Merkel cell carcinoma may be seen more commonly in immunosuppressed patients than in the normal population and that the oncogenic potential of azathioprine and cyclosporin should be borne in mind when prescribing these drugs. PMID- 9390347 TI - Multiple apocrine hidrocystomas of the eyelids. AB - We report a patient with multiple apocrine hidrocystoma (cystadenoma) which was characterized by bilateral distribution of the lesions on the eyelids. The disease is benign, but it may be a marker of two rare inherited disorders, the Schopf-Schulz-Passarge syndrome and a peculiar form of focal dermal hypoplasia. PMID- 9390348 TI - Protease inhibitors protect growth factor activity in chronic wounds. PMID- 9390349 TI - Thermal injury induces both necrosis and apoptosis in rat skin. PMID- 9390351 TI - Subacute cutaneous lupus erythematosus induced by radiation therapy. PMID- 9390352 TI - Milia en plaque arising in discoid lupus erythematosus. PMID- 9390353 TI - Lupus erythematosus profundus with partial C4 deficiency. PMID- 9390354 TI - Erythema elevatum diutinum associated with pulmonary infiltrates. PMID- 9390355 TI - Ehlers-Danlos type IV: non-invasive techniques as diagnostic support. PMID- 9390356 TI - Majocchi's disease in a newborn baby: a familial case. PMID- 9390357 TI - Human orf complicated by bullous pemphigoid. PMID- 9390358 TI - The dapsone hypersensitivity syndrome occurring in a patient with dermatitis herpetiformis. PMID- 9390359 TI - Median canaliform dystrophy following isotretinoin therapy. PMID- 9390360 TI - Weathering nodules of the ear in a young woman. PMID- 9390361 TI - Bowen's disease developing within a Becker's melanosis (Becker's naevus) PMID- 9390362 TI - Tropaeolum majus and contact dermatitis. PMID- 9390363 TI - The Psoriasis Area and Severity Index and alternative approaches for the assessment of severity: persisting areas of confusion. PMID- 9390364 TI - Vision and driving--a literature review and commentary. AB - The visual requirements that currently have to be met those applying for and holding current UK driving licenses for private motor vehicles are discussed, together with the incidence of road accidents and the general scientific and social problems of setting visual standards for driving. Literature relating to the effectiveness of various visual tests in predicting the accident-proneness of individual drivers is reviewed. A striking feature of the data on the age dependence of accidents and visual performance is that although visual performance by most tests steadily declines after early middle age, older drivers have less accidents than their younger counter-parts, whose visual performance is superior. It is concluded that, although correlations between poor vision as assessed by some tests and accident rates can be shown in large samples of drivers, as yet, no single test or combination of tests has been shown to be able to effectively screen out those at risk of accidents without also leading to the disqualification of a substantial number of potentially safe drivers. Thus no change in the present visual requirements is recommended at the present time. PMID- 9390365 TI - Eye movements and reading with simulated visual impairment. AB - The purpose of this study was to investigate the effects of simulated visual impairment on the reading speed and reading eye movements of young, normally sighted observers. Afocal diffusing filters (Ryser occlusion foils) were used to create three levels of impairment and eye movements were recorded using a spectacle-mounted, infra-red limbal reflection system. Reading speed decreased significantly (P < 0.01) as the level of impairment increased. Eye movement analysis revealed the main contributory factors to be increased fixation durations, shorter saccades (resulting in increased numbers of forward saccades per line) and, to a lesser extent, increased time required for page navigation. The results suggest that in order to achieve optimal reading speeds, print size should be at least four times the acuity threshold and that print contrast should be at least twenty times contrast threshold. PMID- 9390366 TI - Efficiency of the Ishihara test for identifying red-green colour deficiency. AB - The Ishihara test is the most widely used screening test for red-green colour deficiency. Results obtained by 401 people with red-green colour deficiency show that the combined sensitivity of the Transformation and Vanishing plates of the 38 plate Edition of the Ishihara plates is 95.5% on eight errors, 97.5% on six errors and 99.0% on three errors. The Hidden digit designs only identified approximately 50% of colour-deficient subjects. The protan/deutan classification plates were found to be more effective for deutans than for protans. No classification was obtained for 18% of protanopes and 3% of deuteranopes who saw neither figure on classification plates; 40% of protanomalous trichromats and 37.5% of deuteranomalous trichromats saw both classification figures and were classified on the relative luminance (clarity) of these figures. The specificity of the Ishihara test was determined in a previous study (Birch and McKeever, 1993) and the results combined with the present data to obtain the overall efficiency of the Ishihara plates for a representative cross section of colour deficient subjects. PMID- 9390367 TI - Comparison of a videokeratoscope and an autokeratometer as predictors of the optimum back surface curves of rigid corneal contact lenses. AB - PURPOSE: To determine the differences in the corneal topography derived using a 16 mire videokerascope and a 2 mire autokeratometer and to examine whether the differences are clinically significant in the contact lens fitting context. METHODS: The right corneas of 20 subjects were measured by an Eyesys videokeratoscope (windows workstation: version 2000 W) and a Topcon Autokeratometer (KR 3500). The corneal vertex radius and p-value were deduced and used to calculate the back surface specifications of a rigid corneal tricurve contact lens design required for an optimal fit on the corneal model. The study was aimed to evaluate the differences in contact lens specifications related to the current British Standards on contact lens tolerances. RESULTS: In general there was good agreement in the lens specification derived from the two instruments. The differences that were present were small and, with the exception of the second back peripheral radius, were within tolerance limits. CONCLUSIONS: The corneal topography was adequately described by the two mire keratometer for the purpose of fitting this particular lens design on the corneas of the 20 subjects examined. PMID- 9390368 TI - The effect of an artificially-elevated intraocular pressure on corneal thickness in Chinese eye. AB - We measured the central corneal thickness and the applanation intraocular pressure (IOP) on 45 Hong Kong Chinese. There was no obvious relationship between these two parameters, as different from other literatures. It could be due to either a limited number of subjects with a high IOP level (only six subjects with IOP > or = 22 mmHg), or Chinese has a thicker central cornea in general. The mean central cornea of our subjects was thicker (566 +/- 36 microns) than some previous findings. Thirty subjects had their intraocular pressure further increased by adopting a 40 degrees head-down posture. Their IOP and topographic corneal thickness were measured again. There was no significant change in the central corneal thickness even though the IOP was elevated by 11.7 mmHg. However the nasal cornea demonstrated a thinning effect (by some 18 microns) during the IOP elevation but it returned to the pre-inverted level after returning to a sitting posture for 5 min. Further investigation with more corneal regions being measured would be valuable to evaluate the in vivo effect of IOP elevation from glaucoma attack on corneal thickness. PMID- 9390369 TI - Some methodological issues in the assessment of the spontaneous eyeblink frequency in man. AB - Previous assessments of spontaneous eyeblink frequency (SEBF) or interblink intervals (IBI) have been made over period of 0.5 to 15 min and average values calculated; the reliability of the methods has not been validated. Video recordings were made of 14 healthy volunteers, aged 20 to 38 years, while silently fixating on a 2 m distant, 35 mm high target under 350 lux illumination and the traces assessed with an event marker. Significant fluctuations in SEBF or IBI were generally observed, but which did not conform to a minute-by-minute periodicity. Time-dependent trends were uncommon, although uncritical pooling or averaging of data can effectively conceal such fluctuations or trends. Correlation's between SEBF and IBI indicate that eyeblink monitoring over at least 3 min is required. Simple averaging calculations are not appropriate because of a high chance of non-Gaussian distribution of data. Modal IBI values correlated well with an adjusted modal calculated SEBF which is thus recommended for further use. PMID- 9390370 TI - Improved computing scheme for measuring eye alignment with Purkinje images I and IV. AB - This study introduces an improved computing scheme for determining eye rotation from Purkinje images I and IV. The original computing scheme systematically underestimated eye rotation. Paraxial raytracing calculations revealed that this error resulted from failure to account for the fact that Purkinje images I and IV fall at different distances behind the cornea. The error could be overcome with a correction factor derived from paraxial raytracing calculations. A series of experiments were carried out to test the validity of this correlation factor, involving exact raytracing calculations as well as measurements on physical model eyes and human eyes. The influence on the correction factor of ocular surface asphericity, accommodation, age and ocular component variations were examined. The new method was also compared to Hirschberg's technique, which makes use of Purkinje image I alone, as a means of screening for strabismus. PMID- 9390371 TI - Graphical representation of visual acuity data. AB - Accurate graphical depiction of data requires that distances between points map precisely to the quantitative differences they represent. In the case of Snellen visual acuity, consecutive line sizes do not generally correspond to equal increments in visual angle, hence scores plotted against axes marked off in equal intervals provide a false representation of the relation between plotted values. A worked example illustrates the nature of this problem and its solution. PMID- 9390372 TI - Active emmetropization--evidence for its existence and ramifications for clinical practice. AB - There is increasing evidence from animal studies in support of the concept of an active emmetropization mechanism which has potentially important clinical ramifications for the management of refractive errors. Recent research into refractive development and emmetropization is reviewed, with emphasis given to work involving the chick, tree shrew and monkey, which represent the three most widely used animal models in this field. The findings of this research are reviewed in a clinical context. Compensatory eye growth responses to focusing errors imposed by lenses represent the most compelling evidence for active emmetropization. These observations are complemented by other evidence showing recovery from induced refractive errors such as form-deprivation myopia. Of the animals listed above, chicks show the most impressive emmetropization, being able to compensate fully (using choroidal and scleral mechanisms) to lens powers ranging from +15 D to -10 D. The range of lens powers eliciting appropriate compensatory responses is narrower in the tree shrew and monkey, and the response patterns generally are also more complex to interpret. These data relate to young animals and together indicate refractive plasticity during development. Extrapolation of these findings to humans predicts that natural emmetropization will be inhibited in neonates by early intervention with prescription lenses, and that refractive correction of myopia will lead to accelerated progression. This convincing evidence for active emmetropization warrants due consideration in developing clinical management strategies for refractive errors. PMID- 9390373 TI - Accommodation and acuity under night-driving illumination levels. AB - Laboratory experiments are described in which the monocular changes in the refractive error and acuity of six young, normal, adult subjects were measured as the field luminance was reduced from approximately 100 to 10(-3) cd/m2. It was found that, at luminance levels equal to those recommended for road lighting (about 1 cd/m2), acuity fell from its photopic value of > or = 6/6 to about 6/9, with little change in the measured refraction. Marked changes in refraction, i.e. night myopia, only started to become manifest when the luminance was further reduced to below about 0.03 cd/m2, much less than that applying under normal night-driving conditions. Direct experiments under street-lighting conditions confirmed the absence of any significant night myopia. It is concluded, therefore, that neural changes, rather than night myopia, normally are responsible for the acuity loss suffered by drivers at night. PMID- 9390374 TI - Patterns of cataract referral in the West Midlands. AB - Optometrists are required to refer patients with any abnormality of the eye, excluding normal changes due to age, to a General Practitioner. It can be difficult to decide when a cataract ceases to be a normal age-related change and becomes an ocular abnormality. This questionnaire-based study examines the criteria adopted by Optometrists and GPs for referring patients with cataract and compares this to the referral criteria suggested by Ophthalmologists. For all professions, the main factors influencing referral are visual acuity, subjective visual impairment and the need to drive. Optometrists and GPs generally refer patients with cataract at a VA level of 6/18-6/24, whilst most Ophthalmologists would be willing to see patients with cataract at a VA of 6/9-6/12. However, Ophthalmologists report that sometimes patients are referred too soon, regardless of their acuity level, because the patient is not yet impaired by their level of vision or does not want surgery. Suggestions for improvements in the referral of cataract are discussed. It is important for Optometrists to make clear whether a letter to a GP is for information only or for referral. If referral is warranted, information in addition to acuity will assist an Ophthalmologist in prioritizing appointments, including age, occupation, brief description of disability and the need to drive without contravening any laws. PMID- 9390375 TI - The AC/A ratio, age and presbyopia. AB - Previous reports concerning the effect of age on the AC/A ratio have been equivocal. Therefore, the present study investigated both the stimulus (AC/As) and response (AC/Ar) ratios using a subjective haploscope-optometer in a relatively large sample of subjects (n = 42) over a wide range of ages (22-65 years). The AC/As showed a small but significant decrease with age (approximately 0.04 delta/D/year). When the older subjects (> 45 years of age) were excluded, however, there was no systematic age effect. The AC/Ar exhibited a small but significant increase with age (approximately 0.08 delta/D/year) for subjects under 45 years of age. However, when the older pre-presbyopes and younger presbyopes (35-44 years of age) were excluded, there was no systematic age effect. In subjects 45 years of age and older, the AC/Ar could not be reliably assessed. This was attributed to physiological and instrumentation noise as a result of the minimal change in accommodative response. In the mid-aged subjects (35-44 years of age), the apparent increase in AC/Ar with age was speculated to be due to neural adaptation of the crosslink gain from the accommodative to the vergence system and/or slight intrusion into the upper non-linear response region of accommodation with the measurements. The finding of AC/Ar constancy with age when mid-aged pre-presbyopes and early presbyopes were excluded supports the non linearity hypothesis, and thus there appears to be no real change in AC/Ar with increased age. The results support the Hess-Gullstrand theory of presbyopia. PMID- 9390376 TI - Pupil size, mean accommodation response and the fluctuations of accommodation. AB - We wished to determine how pupil size and mean accommodation response level interact to influence the fluctuations of accommodation. A dynamic infra-red optometer was used to record accommodation responses while subjects viewed a steady target at two stimulus levels (1.5 and 3 D) through four pupils (1, 2, 4 and 6 mm). It was found for most subjects that the fluctuations of accommodation increase at higher mean accommodation response levels, and small pupils lead to an increase in the low frequency (but not the high frequency) fluctuations of accommodation. The effects of mean accommodation response are independent of pupil size, and the effects of pupil size are independent of mean response level. PMID- 9390377 TI - Fixation disparity and accommodation as a function of viewing distance and prism load. AB - Fixation disparity was measured with dichoptically presented nonius lines at viewing distances of 20, 30, 40, 60, and 100 cm, so that both vergence and accommodation were stimulated adequately as in normal vision. As the viewing distance was shortened, mean fixation disparity changed monotonically from 1 min arc eso (i.e., the eyes converged in front of the target) to 3 min arc exo. The average standard deviation of the psychometric function of fixation disparity, which is a measure of the temporal variability of vergence, was smallest at 100 cm (when fixation disparity was eso) and increased as viewing distance decreased. Fixation disparity itself and the change of fixation disparity with distance differed reliably among subjects with normal binocular vision, but neither was related to the momentary degree of accommodation. Fixation disparity was also measured at a constant distance of 40 cm, but with prisms in front of the eyes that induced the same vergence angles as viewing distances between 20 and 100 cm. The slope of these conventional fixation disparity curves as a function of prism load was generally larger than the slope of fixation disparity as a function of viewing distance (which can be explained by accommodative vergence), but the slopes of the two types of curves were correlated (r = 0.39, P = 0.02, n = 25). PMID- 9390378 TI - Partition of perceived space within the fusional area on the basis of apparent fronto-parallel plane criterion. AB - The relationship between visual space and physical space was investigated by means of the apparent fronto-parallel plane percept. A set of curves in physical space, corresponding to fronto-parallel planes perceived in front or behind a given fixation point, was determined. The curvature at the apex of these curves follows a hyperbolic law that depends on the distance between the perceived plane and the fixation point. Our results are interpreted by distinguishing absolute disparity, which sets the position of the perceived plane, from relative disparity measured by adjusting in this plane eccentric vertical lines. We show that Foley's model, which interprets the shape of the apparent fronto-parallel planes on the mis-evaluation of egocentric distance, works for absolute disparity judgements, but seems to be inadequate for relative disparity judgements in our experimental conditions. PMID- 9390379 TI - Measurement of posterior corneal asphericity on Hong Kong Chinese: a pilot study. AB - The posterior corneal p-value and apical radius of 60 Hong Kong Chinese were assessed. The values were derived based on the information of the anterior corneal topography and the corneal thickness in different regions. The mean posterior corneal apical radius along the horizontal meridian was 6.51 mm (SD +/- 0.40 mm) and the p-value was 0.34 (SD +/- 0.38). The apical radius is greater while the p-value is smaller than a previous study using a similar principle. This may indicate a flatter posterior cornea and greater peripheral flattening in Hong Kong Chinese. No significant difference between the nasal and temporal corneal thickness, nasal and temporal posterior p-value and apical radius was demonstrated. The right and left eyes were also similar in different ocular parameters apart from a smaller anterior corneal p-value on the right eye (R eye: 0.70 +/- 0.13; L eye: 0.67 +/- 0.12), but the difference may not be significant clinically. The method used here is simple and the generation of posterior corneal topography is informative. PMID- 9390380 TI - Double-pass measurements of retinal image quality in monofocal contact lens wearers. AB - The double-pass method is applied to determine the optical image quality in monofocal contact lens (CL) wearers. This is an objective non-invasive technique that permits in vivo testing of the optical performance of CL wearers' eyes. Retinal image quality was measured for three subjects wearing two types of monofocal CLs: a rigid gas permeable (RGP) CL and a soft contact lens (SL), for pupil diameters of 3 mm and 5 mm. Results show the importance of ocular astigmatism regarding retinal image quality. In eyes presenting corneal astigmatism, the best results are obtained when wearing RGP CLs, because the lens compensates the corneal astigmatism. The modulation transfer function (MTF) is considerably smaller when no lens or soft lenses are worn, even for small amounts of astigmatism (0.5 D). When the astigmatism is corrected, the retinal image quality obtained with both types of CLs and with no lens is similar. PMID- 9390381 TI - Proteins of morula-like cells in hemolymph of the giant clam, Tridacna derasa. AB - The morula-like cell, a hemocyte packed with many large (about 3 microns in diameter) electron-dense granules, is found only in the hemolymph of giant clams belonging to the Tridacnidae. To clarify the function of the morula-like cell, we investigated its proteins, especially those found in the large granules. Proteins with molecular weights of 64 kDa, 17 kDa and 7.4 kDa were found to be specific to this type of hemocyte. N-terminal amino acid sequence analysis revealed that the 17-kDa and 7.4-kDa proteins were novel proteins rich in aromatic amino acids. Rabbit polyclonal antibody against a synthetic peptide of the 7.4-kDa protein reacted not only with that protein but also with a larger molecular weight (about 16-kDa) protein in the morula-like cell. Examination of the N-terminal amino acid sequences showed that the 16-kDa protein is distinct from the 17-kDa protein, and Western blot analysis suggested that it is a precursor of the 7.4-kDa protein. The zooxanthellate portion of clam mantle and kidney contained proteins immunoreactive to the antibody, but the azooxanthellate portion of the mantle did not contain any immunoreactive protein. These results suggest that the morula like cells interact with the zooxanthellae. PMID- 9390382 TI - DNA-dependent protein phosphorylation activity in Xenopus is coupled to a Ku-like protein. AB - DNA-dependent protein kinase (DNA-PK) is a nuclear enzyme and functions as a serine/threonine kinase that has been well characterized in both the human and the mouse. The regulatory subunit of DNA-PK is the Ku autoantigen. To demonstrate that a Ku-like protein is present in Xenopus oocytes, we used immunoprecipitation analysis with a monoclonal antibody raised against human Ku antigen and autoimmune serum containing anti-Ku antibodies. Metabolic labeling studies indicate that the Ku-like protein is synthesized mainly in late vitellogenic oocytes. By using a specific peptide substrate for DNA-PK, we demonstrate the activity of a DNA-dependent protein kinase in oocyte extracts. The kinase activity requires the Ku-like protein, since extracts depleted of Ku protein by immunoadsorption with human anti-Ku antibodies fail to demonstrate the DNA dependent phosphorylation activity. The increased enzyme activity in vitellogenic oocytes may be correlated to the increased levels of Ku protein observed in these oocytes compared to the pre- and early vitellogenic oocytes. PMID- 9390384 TI - Circadian rhythms in the lateral eye of the Japanese horseshoe crab. PMID- 9390385 TI - Prediction of maximum allowable retinal slip speed in the fiddler crab, Uca pugilator. PMID- 9390386 TI - Histamine: putative transmitter for lateral inhibition in Limulus eye. PMID- 9390387 TI - Visual performance of horseshoe crabs: role of underwater lighting. PMID- 9390388 TI - UV cutting of MAPs-bound microtubules. PMID- 9390389 TI - Actin bundles in neuronal growth cone observed with the Pol-Scope. PMID- 9390390 TI - Characterization of antibodies to the head and tail domains of squid brain myosin V. PMID- 9390391 TI - Dynamics of GFP-coronin and eupodia in live Dictyostelium observed with real-time confocal optics. PMID- 9390392 TI - Effect of gossypol on Spisula sperm observed with real-time confocal microscopy, polarized light microscopy, and video microscopy. PMID- 9390393 TI - Effect of in vitro culture of mammalian embryos on the architecture of the zona pellucida. PMID- 9390394 TI - Phylogenetic analysis of the 5-aminolevulinate synthase gene. PMID- 9390395 TI - Phylogenetic analysis of olfactory receptor genes from mudpuppy (Necturus maculosus). PMID- 9390396 TI - Genes coding for reverse transcriptase, DNA-directed RNA polymerase, and chitin synthase from the microsporidian Spraguea lophii. PMID- 9390397 TI - Apparatus for measuring steady-state ATP utilization rates of single muscle fibers. PMID- 9390398 TI - Do alpha-crystallins protect catalase against UV damage? PMID- 9390400 TI - Fast voltage-sensitive dye recording of membrane potential changes at multiple sites on an individual nerve cell in the rat cortical slice. PMID- 9390401 TI - Protein solution structure calculations in solution: solvated molecular dynamics refinement of calbindin D9k. AB - The three-dimensional solution structures of proteins determined with NMR-derived constraints are almost always calculated in vacuo. The solution structure of (Ca2+)2-calbindin D9k has been redetermined by new restrained molecular dynamics (MD) calculations that include Ca2+ ions and explicit solvent molecules. Four parallel sets of MD refinements were run to provide accurate comparisons of structures produced in vacuo, in vacuo with Ca2+ ions, and with two different protocols in a solvent bath with Ca2+ ions. The structural ensembles were analyzed in terms of structural definition, molecular energies, packing density, solvent-accessible surface, hydrogen bonds, and the coordination of calcium ions in the two binding loops. Refinement including Ca2+ ions and explicit solvent results in significant improvements in the precision and accuracy of the structure, particularly in the binding loops. These results are consistent with results previously obtained in free MD simulations of proteins in solution and show that the rMD refined NMR-derived solution structures of proteins, especially metalloproteins, can be significantly improved by these strategies. PMID- 9390402 TI - Efficient enzymatic synthesis of 13C,15N-labeled DNA for NMR studies. AB - The power of heteronuclear NMR spectroscopy to study macromolecules and their complexes has been amply demonstrated over the last decade. The obstacle to routinely applying these techniques to the study of DNA has been the synthesis of 13C,15N-labeled DNA. Here we present a simple and efficient method to generate isotope-labeled DNA for NMR studies that is as easy as that for isotope labeling of RNA. The method was used to synthesize a uniformly 13C,15N-labeled 32 nucleotide DNA that binds to human basic fibroblast growth factor with high affinity and specificity. Isotope-edited experiments were applied to the 13C,15N labeled DNA bound to unlabeled protein, and the 13C,15N-labeled DNA was also examined in complex with 15N-labeled protein. The NMR experiments show that the DNA adopts a well-defined stable structure when bound to the protein, and illustrate the potential of 13C,15N-labeled DNA for structural studies of DNA protein complexes. PMID- 9390403 TI - Measurement of diffusion constants for nucleic acids by NMR. AB - Pulsed field-gradient NMR experiments can be used to measure the diffusion constants of nucleic acids. The diffusion constants measured in this way for double-helical DNAs of defined length agree well both with theory and with measurements done using other techniques. When applied to RNAs, this experiment easily distinguishes duplex RNAs from RNA hairpins and thus it can solve one of the perennial problems faced by RNA spectroscopists, i.e. assessing whether their samples are monomeric or not. PMID- 9390404 TI - Three-dimensional structure of the Hck SH2 domain in solution. AB - The hematopoietic cellular kinase (Hck) is a member of the Src family of non receptor protein-tyrosine kinases that is expressed predominantly in granulocytes, monocytes and macrophages. Recent observations suggest that Hck may be activated in HIV-infected macrophages and in chronic myelogenous leukemia cells that express Bcr-Abl. In order to increase our understanding of the structural basis for regulation of Hck activity under normal and pathological conditions, we have solved the solution structure of the uncomplexed Hck SH2 domain using NMR spectroscopy. A novel procedure that uses intraresidue HN-H alpha distances as references for converting NOE intensities into distance restraints has been described. A total of 1757 significant experimental restraints were derived from NMR spectroscopic data including 238 medium-range and 487 long-range distance restraints and 177 torsion angle restraints. These restraints were used in a simulated annealing procedure to generate 20 structures with the program DYANA. Superimposition of residues 5-104 upon the mean coordinate set yielded an average atomic rmsd values of 0.42 +/- 0.08 A for the N,C alpha,C' atoms and 0.81 +/- 0.08 A for all heavy atoms. Rmsd values for those residues in the regions of ordered secondary structure were 0.27 +/- 0.04 A for the N,C alpha,C' atoms and 0.73 +/- 0.06 A for all heavy atoms. PMID- 9390406 TI - The button test: a small scale method using microdialysis cells for assessing protein solubility at concentrations suitable for NMR. AB - A simple method has been developed for screening solution conditions to determine conditions under which a protein is soluble at the high concentrations typically used for NMR spectroscopy. The method employs microdialysis cells or 'buttons'. The low sample volume (5 microliters) required for each microdialysis button permits testing of a wide range of solution conditions and temperatures with high protein concentrations, using a small amount of protein. Following precipitation of several NMR samples of the C-terminal core domain of human TFIIB, the microdialysis button screen facilitated identification of conditions in which precipitation of the TFIIB core domain was eliminated. The microdialysis button method for screening solution conditions is generally applicable and has been used to permit rapid identification of suitable NMR sample solution conditions for proteins involved in transcription and cell adhesion. PMID- 9390405 TI - Conformation of an Shc-derived phosphotyrosine-containing peptide complexed with the Grb2 SH2 domain. AB - We have determined the structure of an Shc-derived phosphotyrosine-containing peptide complexed with Grb2 SH2 based on intra- and intermolecular NOE correlations observed by a series of isotope-filtered NMR experiments using a PFG z-filter. In contrast to an extended conformation of phosphotyrosine-containing peptides bound to Src, Syp and PLC gamma SH2s, the Shc-derived peptide formed a turn at the +1 and +2 positions next to the phosphotyrosine residue. Trp121, located at the EF1 site of Grb2 SH2, blocked the peptide binding in an extended conformation. The present study confirms that each phosphotyrosine-containing peptide binds to the cognate SH2 with a specific conformation, which gives the structural basis for the binding specificity between SH2s and target proteins. PMID- 9390407 TI - High-resolution heteronuclear NMR of human ubiquitin in an aqueous liquid crystalline medium. AB - A mixture of dihexanoyl phosphatidylcholine and dimyristoyl phosphatidylcholine in water forms disc-shaped particles, often referred to as bicelles [Sanders and Schwonek (1992) Biochemistry, 31, 8898-8905]. These adopt an ordered, liquid crystalline phase, which can be maintained at very low concentrations of the bicelles (down to 3% w/v). At this concentration the spacing between individual bicelles, on average, exceeds 300 A. The bicelles are shown to have a negligible effect on the rotational diffusion of ubiquitin as judged by the 15N T1p values of the backbone amides relative to those in isotropic aqueous solution. The protein exhibits a residual degree of alignment which is proportional to the bicelle concentration, and approximately collinear with ubiquitin's rotational diffusion tensor. The degree of alignment obtained offers unique opportunities for studying the protein's structure and dynamics. PMID- 9390409 TI - Assignment of the 1H, 15N and 13C resonances of the calcium-free and calcium bound forms of the first C2-domain of synaptotagmin I. PMID- 9390408 TI - Tritium NMR studies of the human carbonic anhydrase I-benzenesulfonamide complex. AB - Tritium NMR spectroscopy has been used to examine the complex formed by [4 3H]benzenesulfon-amide and human carbonic anhydrase I. The results show that in solution the inhibitor forms a 1:1 complex with the enzyme. A 100-spin computational model of the system, constructed with reference to crystallographic results, was used to interpret tritium relaxation behavior and 3H{1H} NOEs. The analysis shows that the rate of dissociation of the enzyme-sulfonamide complex is 0.35 s-1 and that the aromatic ring of the inhibitor undergoes rapid rotation while complexed. PMID- 9390410 TI - Backbone assignment of double labelled 23.7 kDa phosphoglycerate mutase from Schizosaccharomyces pombe. PMID- 9390411 TI - Sequence-specific assignments of the inner lipoyl domain of human pyruvate dehydrogenase. PMID- 9390412 TI - Human nucleotide excision repair protein XPA: expression and NMR backbone assignments of the 14.7 kDa minimal damaged DNA binding domain (Met98-Phe219). PMID- 9390413 TI - The effects of self-esteem and source credibility on self-denying prophecies. AB - Self-fulfilling prophecies are a well-studied phenomenon. The study of self denying prophecies, however, is rare. Self-denying prophecies shift people's behavior in the direction opposite to the prophecy. The existence of self-denying prophecies was investigated in 222 students. The effects of self-esteem and the source of the prophecy were also investigated. The results suggest that self denying prophecies exist and that self-esteem is an important moderator of self denying prophecies. If managers and industrial/organizational psychologists had an understanding of self-denying prophecies, they might be better able to structure negative performance reviews in a way that could lead to improved employee performance. PMID- 9390414 TI - Psychosocial adjustment in male-to-female transsexuals: an overview of the research evidence. AB - Transsexualism has been defined as an extreme gender dysphoria; it refers to unhappiness with one's biological sex and the desire to have the body of the opposite sex and to be regarded by others as a member of that other sex. Transsexualism is not a common condition, but its prevalence is not yet known. A large number of transsexuals receive hormonal treatment and sex reassignment surgery (SRS). In spite of years of poor quality research, due in part to methodological problems, recent research on surgical outcomes has provided important information. However, psychological research into transsexualism has ignored the cognitive style and psychological functioning of transsexuals, and very little effort has been made to incorporate research findings into the development of psychological treatments to improve the quality of life for transsexuals. PMID- 9390415 TI - Psychophysiological aspects of Tourette's syndrome. AB - Tourette's syndrome (TS), once considered a rare disorder, has been investigated extensively in the last two decades. It is inherited, usually beginning in childhood, and waxes and wanes, usually decreasing in frequency and severity in adolescence and early adulthood. Pharmacotherapy is the usual treatment approach, reducing frequency and severity of symptoms, but it is not a cure and often has side effects. Psychological help for people with TS and their families may be needed for this complex disorder. PMID- 9390416 TI - Assessing the learning processes of black South African students. AB - Data based on responses of 126 male and 201 female 14- and 15-year-old Black South African secondary school students showed the Learning Process Questionnaire (LPQ; Biggs, 1987) to be fairly reliable and factorially valid. Comparison with the LPQ means for like-aged students from Australia and Hong Kong called into question the common assertion that Black South African students are more prone to use superficial learning processes than are Western students. In particular, the South African responses to the LPQ indicated that they were less shallow and more oriented toward achievement in their approach to learning than the Australian students were. PMID- 9390417 TI - Children's perceptions of purposes for studying different subjects in school. AB - Examiners explored 4th and 6th graders' perceptions of studying 6 subjects in school: language, math, science, social studies, music, and visual arts. Participants were asked to indicate whether they agreed or disagreed with each of 12 reasons for studying each of the 6 subjects. Five of the reasons included general educational goals that were applicable to all subjects (cross-curricular goals), 5 included objectives established for a particular subject (subject specific objectives), and 2 represented goals irrelevant to the subject. Both 4th and 6th graders selected mainly subject-specific objectives (e.g., "learn about different rhythms") as reasons for studying music and visual arts. Many children used some of the general cross-curricular goals (e.g., "do well in school in the future"), in addition to the subject-specific goals, to describe why they were studying core subjects: language, math, social studies, and science. PMID- 9390418 TI - Examining the generalizability of the orality-depression link. PMID- 9390419 TI - Cloning and characterization of a cDNA encoding an elicitor of Phytophthora parasitica var. nicotianae that shows cellulose-binding and lectin-like activities. AB - Phytophthora parasitica var. nicotianae produces a 34-kDa glycoprotein elicitor (CBEL) that is localized in the cell wall. A cDNA encoding the protein moiety of this elicitor was cloned and characterized. The deduced amino acid sequence consisted of two direct repeats of a cysteine-rich domain, joined by a Thr/Pro rich region. Although having no general homology with published sequences, the positions of the cysteine residues in the two repeats show a conserved pattern, similar to that of the cellulose-binding domain of fungal glycanases. CBEL did not possess hydrolytic activity on a variety of glycans, but bound to fibrous cellulose and plant cell walls. In addition, it exerted a lectin-like hemagglutinating activity. Infiltration of tobacco leaves (cultivar 46-8) with this molecule elicited necrosis and defense gene expression at 150 nM. Elicitor pretreatment of this tobacco cultivar resulted in protection against subsequent inoculation with an otherwise virulent race of P. parasitica var. nicotianae. All these biological activities were exerted within a low concentration range. This is the first report that a fungal elicitor exhibits cellulose-binding and lectin like activities. The possible implications of such a multifunctional elicitor in plant-microbe interactions are discussed. PMID- 9390420 TI - A regulatory locus, pehSR, controls polygalacturonase production and other virulence functions in Ralstonia solanacearum. AB - We previously identified a locus that regulates production of polygalacturonase (PG), an extracellular plant cell wall-degrading enzyme important in bacterial wilt of plants caused by Ralstonia (Pseudomonas) solanacearum. The DNA sequence of this locus, called pehSR, was determined and two consecutive open reading frames (ORFs) of 1,905 and 1,680 bp were identified. The amino acid sequences predicted to be encoded by these ORFs are similar to those of regulators of pilin synthesis in Pseudomonas aeruginosa and Myxococcus xanthus and to a regulator of flagellin synthesis and adhesion in P. aeruginosa, as well as to other two component regulators of the NtrB/C subfamily. pehSR mutants produced negligible levels of endo-PG activity, while exo-PG activity was reduced by 50%. Northern (RNA) blot analysis showed that PehSR regulates endo-PG expression at the transcriptional level. pehSR mutants grew normally in culture and in planta but were dramatically reduced in virulence; this loss of virulence was substantially greater than that observed for endo-PG structural gene mutants, suggesting that pehSR regulates additional factors important in virulence. Although pehSR mutants were essentially nonmotile, like the wild-type strain, multiple copies of pehSR conferred motility on the bacterium. Reporter gene studies indicated that pehSR expression increased when bacteria grew in plant tissue, and that the pehSR locus was itself negatively regulated by the global virulence gene regulator PhcA. PMID- 9390421 TI - Phenotypic variation in transgenic tobacco expressing mutated geminivirus movement/pathogenicity (BC1) proteins. AB - Tobacco plants were transformed with the movement protein (pathogenicity) gene (BC1) from tomato mottle geminivirus (TMoV), using Agrobacterium-mediated transformation. Different transgenic tobacco lines that expressed high levels of the BC1 protein had phenotypes ranging from plants with severe stunting and leaf mottling (resembling geminivirus symptoms) to plants with no visible symptoms. The sequence data for the BC1 transgene from the transgenic plants with the different phenotypes indicated an association of spontaneously mutated forms of the BC1 gene in the transformed tobacco with phenotype variations. One mutated transgene associated with an asymptomatic phenotype had a major deletion at the C terminus of 119 amino acid residues with a recombination resulting in the addition of 26 amino acid residues of unidentified origin. This asymptomatic, mutated BC1 attenuated the phenotypic expression of the symptomatic BC1 in a tobacco line containing both copies of the BC1 gene. Another mutated form of the BC1 gene amplified from an asymptomatic, multicopy transgenic tobacco plant did not induce symptoms when transiently expressed in tobacco via a virus vector. The symptom attenuation in the transgenic tobacco by the asymptomatic BC1 may involve trans-dominant negative interference. PMID- 9390422 TI - G protein alpha subunit genes control growth, development, and pathogenicity of Magnaporthe grisea. AB - Three G protein alpha subunit genes have been cloned and characterized from Magnaporthe grisea: magA is very similar to CPG-2 of Cryphonectria parasitica; magB is virtually identical to CPG-1 of Cryphonectria parasitica, to gna1 of Neurospora crassa, and to fadA of Emericella nidulans; and magC is most similar to gna2 of Neurospora crassa. Homologous recombination resulting in targeted deletion of magA had no effect on vegetative growth, conidiation, or appressorium formation. Deletion of magC reduced conidiation, but did not affect vegetative growth or appressorium formation. However, disruption of magB significantly reduced vegetative growth, conidiation, and appressorium formation. magB- transformants, unlike magA- and magC- transformants, exhibited a reduced ability to infect and colonize susceptible rice leaves. G protein alpha subunit genes are required for M. grisea mating. magB- transformants failed to form perithecia, whereas magA- and magC- transformants did not produce mature asci. These results suggest that G protein alpha subunit genes are involved in signal transduction pathways in M. grisea that control vegetative growth, conidiation, conidium attachment, appressorium formation, mating, and pathogenicity. PMID- 9390423 TI - Iron-dependent transcription of the regulatory gene ros of Agrobacterium radiobacter. AB - Transcription of the regulatory gene ros of Agrobacterium radiobacter requires growth in the presence of Fe although this regulation was not mediated by ros itself. The ros gene repressed its own transcription, independently of the Fe status of the growth media. It was shown that the two cysteine residues in the Ros protein were essential for the complementation of the exopolysaccharide synthesis defect of ros mutant strains. It was found that the mutation in one exo mutant strain that was complemented both by ros and by the "structural" exoY gene was not, in fact, in ros but is in some other, unknown gene. The two cysteines were also essential for the correction of this mutant. This mutant affected the expression of exoY but not of ros. PMID- 9390424 TI - Transgenic Arabidopsis lines expressing gene VI from cauliflower mosaic virus variants exhibit a range of symptom-like phenotypes and accumulate inclusion bodies. AB - Gene VI of cauliflower mosaic virus (CaMV) is an important determinant of symptom expression during infection. We have constructed a series of transgenic Arabidopsis lines that express gene VI protein (P6) from two CaMV isolates (Bari 1 and Cabb B-JI) that cause mild and severe symptoms, respectively, in Arabidopsis, and from a recombinant virus (Baji-31) with a hybrid gene VI that causes very severe symptoms. From 41 transgenic lines analyzed, 17 showed symptom like phenotypes that ranged from mild vein chlorosis to severe chlorosis and stunting. P6 levels in transgenic lines varied from undetectable in the lowest expressors to levels greater than those in CaMV-infected plants. There was a strong correlation between phenotype severity and the level of P6, and with the gene VI origin in the order, Baji-31 > B-JI > Bari-1. This was similar to symptom severity in Arabidopsis infected with the respective CaMV variant. We also found that transgenic P6 accumulated in inclusion bodies that were similar to those found in infected plants but lacking virions. We conclude that expression of P6, in the absence of virus replication, elicits a subset of the host symptom responses normally observed during infection and that the level, sequence, and possibly the form of P6 are important in potentiating the process. PMID- 9390425 TI - Starvation-induced genes of the tomato pathogen Cladosporium fulvum are also induced during growth in planta. AB - The pathogenicity of fungal pathogens is presumably dependent on genes that are expressed during infection. In order to isolate such genes from the tomato pathogen Cladosporium fulvum, and to test the hypothesis that starvation-induced genes are also plant induced, a cDNA library was prepared from mycelia grown in a defined medium and then transferred to a starvation medium. The library was then screened with cDNA prepared from starved and replete fungal mycelium. Five unique, differentially expressed cDNAs were isolated from 1,000 clones screened. Northern (RNA) hybridization confirmed that all five were starvation induced. Interestingly, all five were also found to be plant induced. The identity of two of the clones was indicated by partial DNA sequencing as alcohol and aldehyde dehydrogenase. The observed correlation between starvation induction and plant induction in discussed. PMID- 9390426 TI - Diverse amino acid residues function within the type 1 peroxisomal targeting signal. Implications for the role of accessory residues upstream of the type 1 peroxisomal targeting signal. AB - The purpose of this study was to determine whether the plant type 1 peroxisomal targeting signal (PTS1) utilizes amino acid residues that do not strictly adhere to the serine-lysine-leucine (SKL) motif (small-basic-hydrophobic residues). Selected residues were appended to the C terminus of chloramphenicol acetyltransferase (CAT) and were tested for their ability to target CAT fusion proteins to glyoxysomes in tobacco (Nicotiana tabacum L.) cv Bright Yellow 2 suspension-cultured cells. CAT was redirected from the cytosol into glyoxysomes by a wide range of residues, i.e. A/C/G/S/T-H/K/ L/N/R-I/L/M/Y. Although L and N at the -2 position (-SLL, -ANL) do not conform to the SKL motif, both functioned, but in a temporally less-efficient manner. Other SKL divergent residues, however, did not target CAT to glyoxysomes, i.e. F or P at the -3 position (-FKL, -PKL), S or T at the -2 position (-SSI, STL), or D at the -1 position (-SKD). The targeting inefficiency of CAT-ANL could be ameliorated when K was included at the -4 position (-KANL). In summary, the plant PTS1 mostly conforms to the SKL motif. For those PTS1s that possess nonconforming residue(s), other residues upstream of the PTS1 appear to function as accessory sequences that enhance the temporal efficiency of peroxisomal targeting. PMID- 9390427 TI - bor1-1, an Arabidopsis thaliana mutant that requires a high level of boron. AB - bor1-1 (high boron requiring), an Arabidopsis thaliana mutant that requires a high level of B, was isolated. When the B concentration in the medium was reduced to 3 microM, the expansion of rosette leaves was severely affected in bor1-1 but not in wild-type plants. In a medium containing 30 microM B the mutant grew normally but showed female sterility, whereas the wild type was able to set seeds. These defects of the bor1-1 mutant were not detected with supplementation of 100 microM B. In vivo concentrations of B in bor1-1 mutants were lower than those of the wild type, especially in the inflorescence stems. Tracer experiments using 10B suggested that the mutant has defects in uptake and/or translocation of B. The mutation was mapped on the lower arm of chromosome 2. PMID- 9390428 TI - Gene fusions of signal sequences with a modified beta-glucuronidase gene results in retention of the beta-glucuronidase protein in the secretory pathway/plasma membrane. AB - Signal sequences and endoplasmic reticulum (ER) retention signals are known to play central roles in targeting and translocation in the secretory pathway, but molecular aspects about their involvement are poorly understood. We tested the effectiveness of deduced signal sequences from various genes (hydroxyproline-rich glycoprotein [HRGP] from Phaseolus vulgaris; Serpin from Manduca sexta) to direct a modified beta-glucuronidase (GUS) protein into the secretory pathway in transgenic tobacco (Nicotiana tabacum L.). The reporter protein was not secreted to the cell wall/extracellular space as monitored using extracellular fluid analysis (low- or high-ionic-strength conditions) but occurred in membranes with a density of 1.16 to 1.20 g/mL. Membrane-bound GUS equilibrated with the plasma membrane (PM) and the ER on linear sucrose gradients with or without ethylenediaminetetraacetic acid, suggesting that GUS associates with the ER and the PM. Confocal microscopy of fixed cultured cells prepared from GUS control and HRGP signal peptide (SP)-GUS-expressing plants suggested only cytosolic localization in GUS-expressing plants but substantial peripheral localization in HRGP SP-GUS plants, which is consistent with GUS being associated with the PM. Aqueous two-phase partitioning of microsomal membranes from HRGP SP-GUS and Serpin SP-GUS transgenic leaves also indicated that GUS activity was enriched in the ER and the PM. These observations, together with hydrophobic moment plot analysis, suggest that properties of the SP-GUS protein result in its retention in the secretory pathway and PM. PMID- 9390429 TI - Developmental and hormonal regulation of the arabidopsis CER2 gene that codes for a nuclear-localized protein required for the normal accumulation of cuticular waxes. AB - The previously cloned CER2 gene is required for the normal accumulation of cuticular waxes and encodes a novel protein. Earlier reports suggested that the CER2 protein is either a membrane-bound component of the fatty acid elongase complex or a regulatory protein. Cell fractionation and immunoblot analyses using polyclonal antibodies raised against a chemically synthesized peptide with a sequence based on the predicted CER2 protein sequence have demonstrated that the 47-kD CER2 protein is soluble and nuclear localized. These results are consistent with CER2 being a regulatory protein. Detailed studies of plants harboring a CER2 promoter/GUS transgene (CER2-GUS), in combination with immunoblot analyses, revealed that CER2 is expressed and the CER2 protein accumulates in a variety of organs and cell types. Expression is highest early in the development of these organs and is epidermis specific in most tissues. In agreement with the activity of the CER2 promoter in hypocotyls, cuticular wax accumulates on this organ in a CER2-dependent fashion. In leaves CER2 expression is confined to the guard cells, trichomes, and petioles. However, application of the cytokinin 6 benzylaminopurine induces ectopic expression of CER2-GUS in all cell types of leaves that emerge following treatment. PMID- 9390430 TI - Heat treatment results in a loss of transgene-encoded activities in several tobacco lines. AB - Heat treatment (37 degrees C) of transgenic tobacco (Nicotiana tabacum) plants led to a reversible reduction or complete loss of transgene-encoded activities in about 40% of 10 independent transformants carrying the luciferase-coding region fused to the 355 cauliflower mosaic virus or the soybean small subunit promoter and the nopaline synthase promoter driving the neomycin phosphotransferase gene, whereas the other lines had temperature-tolerant activities. Temperature sensitivity or tolerance of transgene-encoded activities was heritable. In some of the lines, temperature sensitivity of the transgene-encoded activities depended on the stage of development, occurring in either seedlings (40% luciferase and 50% neomycin phosphotransferase) or adult plants (both 40%). The phenomenon did not correlate with copy numbers or the homo- or hemizygous state of the transgenes. In lines harboring a temperature-sensitive luciferase activity, reduction of bioluminescence was observed after 2 to 3 h at 37 degrees C. Activity was regained after 2 h of subsequent cultivation at 25 degrees C. Irrespective of the reaction to the heat treatment, the level of luciferase RNA was slightly increased at 37 degrees C. Only in lines showing temperature sensitivity of transgene-encoded activities was the amount of luciferase and neomycin phosphotransferase strongly reduced. In sterile culture, heat treatment for 15 d did not cause visible damage or changes in plant morphology. In all plants tested a slight induction of the heat-shock response was observed at 37 degrees C. PMID- 9390431 TI - Starches from A to C. Chlamydomonas reinhardtii as a model microbial system to investigate the biosynthesis of the plant amylopectin crystal. AB - Wide-angle powder x-ray diffraction analysis was carried out on starch extracted from wild-type and mutant Chlamydomonas reinhardtii cells. Strains containing no defective starch synthases as well as mutants carrying a disrupted granule-bound starch synthase structural gene displayed the A type of diffraction pattern with a high degree of crystallinity. Mutants carrying a defect for the major soluble starch synthase (SSS), SSS II, were characterized by a switch to the B type of diffraction pattern with very low crystallinity. Mutant strains carrying SSS I as the only glucan elongation enzyme regained some of their crystallinity but switched to the C type of diffraction pattern. Differential scanning calorimetry analysis correlated tightly with the x-ray diffraction results. Together with the electron microscopy analyses, these results establish C. reinhardtii as a microbial model system displaying all aspects of cereal starch synthesis and structure. We further show that SSS II is the major enzyme involved in the synthesis of crystalline structures in starch and demonstrate that SSS I alone builds a new type of amylopectin structure. PMID- 9390432 TI - Differential patterns of expression of the Arabidopsis PHYB, PHYD, and PHYE phytochrome genes. AB - The Arabidopsis thaliana phyB, phyD, and phyE phytochrome apoproteins show higher amino acid sequence similarity to each other than to phyA or phyC, they are the most recently evolved members of this photoreceptor family, and they may interact in regulating photomorphogenesis. The expression patterns of translational fusions of the 5' upstream regions of the PHYB, PHYD, and PHYE genes to the beta glucuronidase (GUS) coding sequence were compared. PD-GUS and PE-GUS fusions were 5- to 10-fold less active than a PB-GUS fusion, but all three promoter regions drove expression of the reporter gene in all stages of the plant's life cycle. Over the first 10 d of seedling growth, the PHYB and PHYD promoters were more active in the dark than in the light, whereas the opposite was true of the PHYE promoter. Unlike the PB-GUS construct, which was expressed in most parts of seedlings and mature plants, the PD-GUS and PE-GUS transgenes showed differential expression, notably in leaves, flower organs, and root tips. Tissue sections showed that the three promoters are coexpressed in at least some leaf cells. Hence, the PHYB, PHYD, and PHYE genes differ in expression pattern but these patterns overlap and interaction of these receptor forms within individual cells is possible. PMID- 9390433 TI - The COW1 locus of arabidopsis acts after RHD2, and in parallel with RHD3 and TIP1, to determine the shape, rate of elongation, and number of root hairs produced from each site of hair formation. AB - Two recessive mutant alleles at CAN OF WORMS1 (COW1), a new locus involved in root hair morphogenesis, have been identified in Arabidopsis thaliana L. Heynh. Root hairs on Cow1- mutants are short and wide and occasionally formed as pairs at a single site of hair formation. The COW1 locus maps to chromosome 4. Root hairs on Cow1- plants form in the usual positions, suggesting that the phenotype is not the result of abnormal positional signals. Root hairs on Cow1- roots begin hair formation normally, forming a small bulge, or root hair initiation site, of normal size and shape and in the usual position on the hair-forming cell. However, when Cow1- root hairs start to elongate by tip growth, abnormalities in the shape and elongation rate of the hairs become apparent. Genetic evidence from double-mutant analysis of cow1-1 and other loci involved in root hair development supports our conclusion that COW1 is required during root hair elongation. PMID- 9390434 TI - The characterization of plasma membrane-bound tubulin of cauliflower using Triton X-114 fractionation. AB - The cortical microtubules determine how cellulose microfibrils are deposited in the plant cell wall and are thus important for the control of cell expansion. To understand how microtubules can control microfibril deposition, the components that link the microtubules to the plasma membrane (PM) of plant cells must be isolated. To obtain information on the properties of the tubulin-membrane associations, cauliflower (Brassica oleracea) PM was subjected to Triton X-114 fractionation, and the distribution of alpha- and beta-tubulin was analyzed using immunoblotting. Approximately one-half of the PM-associated tubulin was solubilized by Triton X-114 and 10 to 15% of both alpha- and beta-tubulin was recovered in the detergent phase (indicative of hydrophobic properties) and 30 to 40% was recovered in the aqueous phase. The hydrophobic tubulin could be released from the membrane by high pH extraction with preserved hydrophobicity. A large part of the PM-associated tubulin was found in the Triton-insoluble fraction. When this insoluble material was extracted a second time, a substantial amount of hydrophobic tubulin was released if the salt concentration was increased, suggesting that the hydrophobic tubulin was linked to a high-salt-sensitive protein aggregate that probably includes other components of the cytoskeleton. The hydrophobicity of a fraction of PM-associated tubulin could reflect a direct or indirect interaction of this tubulin with the lipid bilayer or with an integral membrane protein and may represent the anchoring of the cortical microtubules to the PM, a key element in the regulation of cell expansion. PMID- 9390435 TI - Characterization of new gibberellin-responsive semidwarf mutants of arabidopsis. AB - Chemical mutagenesis of Arabidopsis thaliana (L.) Heynh. yielded four semidwarf mutants, all of which appeared to be gibberellin (GA)-biosynthesis mutants. All four had atypical response profiles to C20-GAs, suggesting that each had impaired 20-oxidation. One mutant, 11.2, was shown to be allelic to ga5 and has been named ga5-2. It had altered metabolism of [14C]GA15 relative to that in wild-type plants and undetectable levels of C19-GAs in young stems, consistent with the known function of GA5 as a stem-expressed GA 20-oxidase. Two mutants (2.1 and 10.3), which had very short inflorescences and siliques, were allelic to each other but not to the known GA-responding mutants, ga1 to ga5. The locus defined by these two mutations is provisionally named GA6 and is purported to encode an inflorescence- and silique-expressed GA 20-oxidase. A double mutant, ga5-2 ga6-2, had an extreme dwarf phenotype with very short siliques. The fourth mutation, 1.1, gave a phenotype like ga5, but was not allelic to any of the known ga mutations. It has not yet been given a gene symbol pending further studies. PMID- 9390436 TI - Differential expression of chitinases in Vitis vinifera L. responding to systemic acquired resistance activators or fungal challenge. AB - The concept of systemic acquired resistance (SAR) enables a novel approach to crop protection, and particular pathogenesis-related proteins, i.e. an acidic chitinase, have been classified as markers of the SAR response. Basic class I (VCHIT1b) and a class III (VCH3) chitinase cDNAs were cloned from cultured Vitis vinifera L. cv Pinot Noir cells and used to probe the induction response of grapevine cells to salicylic acid or yeast elicitor. Furthermore, the cells were treated with the commercial SAR activators 2,6-dichloroiso-nicotinic acid or benzo(1,2,3)-thiadiazole-7-carbothioic acid S-methyl ester. Elicitor or salicylic acid induced both VCHIT1b and VCH3 transcript abundances, whereas 2,6-dichloroiso nicotinic acid or benzo(1,2,3)-thiadiazole-7-carbothioic acid S-methyl ester enhanced exclusively the expression of VCH3. To assess the systemic sensation of chitinase expression, single leaves of Vitis vinifera L. cv Pinot Noir or Vitis rupestris plants were inoculated with Plasmopara viticola spore suspensions, and the VCH3 and VCHIT1b mRNA amounts in the infected versus the adjacent, healthy leaf were monitored. Two VCH3 mRNA maxima were observed 2 and 6 d postinoculation in the infected, susceptible V. vinifera tissue, whereas in the healthy leaf the transcript increased from low levels d 2 postinoculation to prominent levels d 6 to 8 postinoculation. The level of VCH3 mRNA increased also over 4 d in the inoculated, resistant V. rupestris tissue. However, necrotic spots rapidly limited the infection, and the VCH3 transcript was undetectable in the upper stage, healthy leaf. The expression of VCHIT1b remained negligible under either experimental condition. Overall, the results suggest that the selective expression of VCH3 might be a reliable indicator of the SAR response in V. vinifera L. PMID- 9390437 TI - Characterization and expression of caffeoyl-coenzyme A 3-O-methyltransferase proposed for the induced resistance response of Vitis vinifera L. AB - Cell-suspension cultures of Vitis vinifera L. cv Pinot Noir accumulated resveratrol upon fungal elicitation, and the activity of S-adenosyl-L methionine:trans-caffeoyl-coenzyme A 3-O-methyl-transferase (CCoAOMT), yielding feruloyl-CoA, increased to a transient maximum at 12 to 15 h. CCoAOMT cDNA was cloned from the elicited cells and was shown to encode a polypeptide highly homologous to CCoAOMTs from cells of Petroselinum species or Zinnia species. The expression of the cDNA in Escherichia coli revealed that grapevine CCoAOMT methylates both caffeoyl- and 5-hydroxyferuloyl-coenzyme A and is probably involved in phenolic esterification and lignification. Commercial plant activators induce the disease-resistance response of test plants and are considered to mimic the action of salicylic acid. Among these chemicals, 2,6 dichloroisonicotinic acid and benzo(1,2,3)-thiadiazole-7-carbothioic acid S methyl ester provoke systemic acquired resistance (SAR) and were also shown to induce the expression of class III chitinase in grapevine. The SAR response is classified by an unchanged phenotype of tissues, but the mechanistic basis is unknown. Treatment of the cultured V. vinifera cells with either fungal elicitor or low concentrations of salicylic acid and 2,6-dichloroisonicotinic acid, respectively, raised the CCoAOMT or stilbene synthase transcript abundance, suggesting that grapevine is capable of the SAR response, whereas benzo(1,2,3) thiadiazole-7-carbothioic acid S-methyl ester was ineffective. The data imply for the first time (to our knowledge) that the expression of phenyl-propanoid genes in grapevine is induced by SAR activators without phenotypic consequences and suggest a role for CCoAOMT and stilbene synthase in the disease-resistance response leading beyond the level of pathogenesis-related proteins as markers of the SAR. PMID- 9390438 TI - Induction of 12-oxo-phytodienoic acid in wounded plants and elicited plant cell cultures. AB - Jasmonic acid (JA) is rapidly biosynthesized from alpha-linolenic acid in plants upon contact with pathogens or wounding, and triggers gene activation, leading to the synthesis of defensive secondary metabolites and proteins. Despite the recent finding that its precursor, 12-oxo-phytodienoic acid (PDA), is a more powerful inducer of gene activation, interest has focused so far almost exclusively on JA. A validated negative chemical ionization-gas chromatography-mass spectrometry method has been developed that allows the simultaneous quantification of endogenous 12-oxo-PDA and JA in plant tissues. In six out of eight plant species tested maximal levels of 12-oxo-PDA exceeded peak levels of JA by approximately 3 to 5-fold after elicitation with a yeast cell wall preparation or when plants were wounded. These experiments support the hypothesis that 12-oxo-PDA acts as the predominant jasmonate signal in most plants, whereas JA remains an active metabolite of its precursor. Furthermore, JA but not 12-oxo-PDA was shown to be secreted into the medium from cultured plant cells, suggesting that JA may also act as an intercellular signal. PMID- 9390439 TI - Expression of a vacuolar protein (VP24) in anthocyanin-producing cells of sweet potato in suspension culture. AB - VP24, an abundant protein of 24 kD, was found to accumulate in the anthocyanin containing vacuoles of cells of sweet potato (Ipomoea batatas) in suspension culture. Light-induced expression of VP24 was analyzed by immunoblotting in three different cell lines that produced anthocyanins at different rates. The expression of VP24 was closely correlated with the accumulation of anthocyanin in these cell lines. Immunocytochemical detection of VP24 with specific antibodies on thin sections showed that VP24 was localized in the intravacuolar pigmented globules (cyanoplasts) in the anthocyanin-containing vacuoles and not in the tonoplast. No VP24 immunogold labeling was detected in the vacuoles of the cell line that does not produce anthocyanin. We suggest that VP24 may be involved in the formation of the cyanoplast via an interaction with anthocyanin, and that it may play an important role in the trapping in vacuoles of large amounts of anthocyanins that have been transported into these vacuoles. PMID- 9390440 TI - Characterization of maize elongation factor 1A and its relationship to protein quality in the endosperm. AB - The protein synthesis elongation factor 1A (eEF1A) is a multifunctional protein in eukaryotic cells. In maize (Zea mays L.) endosperm eEF1A co-localizes with actin around protein bodies, and its accumulation is highly correlated with the protein-bound lysine (Lys) content. We purified eEF1A from maize kernels by ammonium sulfate precipitation, ion-exchange, and chromatofocusing. The identify of the purified protein was confirmed by microsequencing of an endoproteinase glutamic acid-C fragment and by its ability to bundle actin. Using purified eEF1A as a standard, we found that this protein contributes 0.4% of the total protein in W64A+ endosperm and approximately 1% of the protein in W64Ao2. Because eEF1A contains 10% Lys, it accounts for 2.2% of the total Lys in W64A+ and 2.3% of the Lys in W64Ao2. However, its concentration predicts 90% of the Lys found in endosperm proteins of both genotypes, indicating that eEF1A is a key component of the group of proteins that determines the nutritional quality of the grain. This notion is further supported by the fact that in floury2, another high-Lys mutant, the content of eEF1A increases with the dosage of the floury2 gene. These data provide the biochemical basis for further investigation of the relationship between eEF1A content and the nutritional quality of cereals. PMID- 9390441 TI - LHT1, a lysine- and histidine-specific amino acid transporter in arabidopsis. AB - We have identified a new amino acid transporter from the Arabidopsis thaliana expressed sequence tag cDNA collection by functional complementation of a yeast amino acid transport mutant. Transport analysis of the expressed protein in yeast shows that it is a high-affinity transporter for both lysine (Lys) and histidine with Michaelis constant values of 175 and 400 microM, respectively. This transporter (LHT1, lysine histidine transporter) has little affinity for arginine when measured directly in uptake experiments or indirectly with substrate competition. The cDNA is 1.7 kb with an open reading frame that codes for a protein with 446 amino acids and a calculated molecular mass of 50.5 kD. Hydropathy analysis shows that LHT1 is an integral membrane protein with 9 to 10 putative membrane-spanning domains. Southern-blot analysis suggests that LHT1 is a single-copy gene in the Arabidopsis genome. RNA gel-blot analysis shows that this transporter is present in all tissues, with the strongest expression in young leaves, flowers, and siliques. Wholemount, in situ hybridization revealed that expression is further localized on the surface of roots in young seedlings and in pollen. Overall, LHT1 belongs to a new class of amino acid transporter that is specific for Lys and histidine, and, given its substrate specificity, it has significant promise as a tool for improving the Lys content of Lys-deficient grains. PMID- 9390442 TI - Analysis of wild-type and mutant plant nitrate reductase expressed in the methylotrophic yeast Pichia pastoris. AB - Recombinant Arabidopsis thaliana NADH:nitrate reductase (NR; EC 1.6.6.1) was produced in the methylotrophic yeast Pichia pastoris and purified to near electrophoretic homogeneity. Purified enzyme had the spectral and kinetic properties typical of highly purified NR from natural plant sources. Site directed mutagenesis altering several key residues and regions was carried out, and the mutant enzyme forms were expressed in P. pastoris. When the invariant cysteine residue, cysteine-191, in the molybdo-pterin region of the A. thaliana NIA2 protein was replaced with serine or alanine, the NR protein was still produced but was inactive, showing that this residue is essential for enzyme activity. Deletions or substitutions of the conserved N terminus of NR retained activity and the ability to be inactivated in vitro when incubated with ATP. Enzyme with a histidine sequence appended to the N terminus was still active and was easily purified using metal-chelate affinity chromatography. These results demonstrate that P. pastoris is a useful and reliable system for producing recombinant holo-NR from plants. PMID- 9390443 TI - Changes in the composition of the photosynthetic apparatus in the galactolipid deficient dgd1 mutant of Arabidopsis thaliana. AB - The glycerolipid digalactosyl diacylglycerol (DGDG) is exclusively associated with photosynthetic membranes and thus may play a role in the proper assembly and maintenance of the photosynthetic apparatus. Here we employ a genetic approach based on the dgd1 mutant of Arabidopsis thaliana to investigate the function of DGDG in thylakoid membranes. The primary defect in the genetically well characterized dgd1 mutant resulted in a 90% reduction of the DGDG content. The mutant showed a decreased photosystem II (PSII) to photosystem I ratio. In vivo room- and low-temperature (77 K) chlorophyll fluorescence measurements with thylakoid preparations are in agreement with a drastically altered excitation energy allocation to the reaction centers. Quantification of pigment-binding apoproteins and pigments supports an altered stoichiometry of individual pigment protein complexes in the mutant. Most strikingly, an increase in the amount of peripheral light-harvesting complexes of PSII relative to the inner antenna complexes and the PSII reaction center/core complexes was observed. Regardless of the severe alterations in thylakoid organization, photosynthetic oxygen evolution was virtually not compromised in dgd1 mutant leaves. PMID- 9390444 TI - A ubiquitous plant housekeeping gene, PAP, encodes a major protein component of bell pepper chromoplasts. AB - We have isolated a cDNA (PAP) corresponding to a single nuclear gene that encodes an approximately 30-kD major protein of bell pepper (Capsicum annuum L.) fruit chromoplasts. RNA and protein analyses revealed that, although at a low level, this gene is also expressed in every organ of the plant, the amount of the corresponding transcript and protein dramatically increasing in the latter stages of fruit development. Western-blot and immunocytochemical analyses of purified chloroplasts from leaves and fruits and of chromoplasts from red fruits showed that the encoded protein is the major component of plastoglobules and fibrils and is localized on the outer surface of these lipid structures. Analyses of PAP in plants belonging to different taxa revealed that it is expressed and highly conserved in both monocotyledonous and dicotyledonous plants. The presence of the protein in plastids not differentiating into chromoplasts indicates that PAP is expressed irrespective of the ontogeny of various plastid lines. In light of our results and since the encoded protein, identical to that previously named ChrB or fibrillin, is present in plastoglobules from several species and accumulates in the fibrils of bell pepper chromoplast, we propose to designate it as a plastid lipid-associated protein. PMID- 9390445 TI - Expression and characterization of pea chloroplastic glyceraldehyde-3-phosphate dehydrogenase composed of only the B-subunit. AB - A cDNA fragment coding for the pea (Pisum sativum L.) chloroplastic glyceraldehyde-3-P dehydrogenase (EC 1.2.1.13) B-subunit and a truncated form corresponding in length to the A-subunit have been cloned into an expression vector, expressed in the absence of the A-subunit in a gap- Escherichia coli strain, purified, and studied. Like the isolated enzyme from higher plant chloroplasts, the recombinant enzymes have dual specificity for NADPH and NADH. The recombinant glyceraldehyde-3-P dehydrogenases have the same optimal pH as the enzyme isolated from pea chloroplasts. Like the native chloroplast enzyme, the recombinant B-subunit has a marked tendency to form large aggregates, whereas the truncated B-subunit exists as the tetramer. The recombinant B-subunit glyceraldehyde 3-P dehydrogenase is more sensitive to dithiothreitol than its truncated form. It seems likely that a different pair of cysteines is responsible for the redox sensitivity of the activity of the enzyme composed of B-subunits than the cysteine residues implicated in the modulation of the activity of the enzyme composed of A-subunits by previous work in this laboratory. PMID- 9390447 TI - Rice hemoglobins. Gene cloning, analysis, and O2-binding kinetics of a recombinant protein synthesized in Escherichia coli. AB - Although nonsymbiotic hemoglobins (Hbs) are found in different tissues of dicots and monocots, very little is known about hb genes in monocots and the function of Hbs in nonsymbiotic tissues. We report the cloning and analysis of two rice (Oryza sativa L.) hb genes, hb1 and hb2, that code for plant Hbs. Rice hb1 and hb2 genes contain four exons and three introns, as with all of the known plant hb genes. At least three copies of the hb gene were detected in rice DNA, and analysis of gene expression shows that hb1 and hb2 are expressed in leaves but only hb1 is expressed in roots. A cDNA for rice Hb1 was expressed in Escherichia coli, and the recombinant Hb (rHb1) shows an unusually high affinity for O2 because of a very low dissociation constant. The absorbance spectra of the ferric and deoxyferrous rHb1 indicate that, in contrast to symbiotic Hbs, a distal ligand is coordinated to the ligand-binding site. Mutation of the distal His demonstrates that this residue coordinates the heme Fe of ferric and deoxyferrous rHb1 and stabilizes O2 in oxy-rHb1. The biochemical properties of rice rHb1 suggest that this protein probably does not function to facilitate the diffusion of O2. PMID- 9390449 TI - The electronic Plant Gene Register. PMID- 9390448 TI - L-ascorbic acid metabolism in the ascorbate-deficient arabidopsis mutant vtc1. AB - The biosynthesis of L-ascorbic acid (vitamin C) is not well understood in plants. The ozone-sensitive Arabidopsis thaliana mutant vitamin c-1 (vtc1; formerly known as soz1) is deficient in ascorbic acid, accumulating approximately 30% of wild type levels. This deficiency could result from elevated catabolism or decreased biosynthesis. No differences that could account for the deficiency were found in the activities of enzymes that catalyze the oxidation or reduction of ascorbic acid. The absolute rate of ascorbic acid turnover is actually less in vtc1 than in wild type; however, the turnover rate relative to the pool of ascorbic acid is not significantly different. The results from [U-14C]Glc labeling experiments suggest that the deficiency is the result of a biosynthetic defect: less L [14C]ascorbic acid as a percentage of total soluble 14C accumulates in vtc1 than in wild type. The feeding of two putative biosynthetic intermediates, D-glucosone and L-sorbosone, had no positive effect on ascorbic acid levels in either genotype. The vtc1 defect does not appear to be the result of a deficiency in L galactono-1,4-lactone dehydrogenase, an enzyme able to convert L-galactono-1,4 lactone to ascorbic acid. PMID- 9390450 TI - Potato pulp: microbiological characterization, physical modification, and application of this agricultural waste product. AB - Potato pulp, one of the agricultural waste products obtained in high quantities during starch production, contains starch, cellulose, hemicelluloses, pectin, proteins, free amino acids and salts. It exhibits physical and physicochemical properties of a typical colloid. It is mainly used, in a dried and pelleted form, as cattle feed. Its autochthonic microbial flora (bacteria, fungi) was identified and studied with a view towards the degradative potential of the microorganisms and ways of conserving the pulp for subsequent technical applications; 33 isolates (28 bacteria, 4 fungi, 1 yeast), belonging to 15 genera were characterized. Biological conservation was possible at very low oxygen pressure, brought about by the autochthonic anaerobic microorganisms causing acidification. Chemical conservation was achieved with sorbic acid. By treatment with hot water vapour under pressure (autoclaving), followed by a pressure release procedure, intact cells in the pulp (both potato cells and microorganisms, not spores) were destroyed, and their contents and wall fragments were set free. This process resulted in low drying costs and was a prerequisite for the production of a powder that can be used as glue or as animal feed. PMID- 9390451 TI - Biodegradation of the pesticide 4,6-dinitro-ortho-cresol by microorganisms in batch cultures and in fixed-bed column reactors. AB - A mixed culture of microorganisms able to utilize 4,6-dinitro-ortho-cresol (DNOC) as the sole source of carbon, nitrogen and energy was isolated from soil contaminated with pesticides and from activated sludge. DNOC was decomposed aerobically in batch cultures as well as in fixed-bed column reactors. Between 65% and 84% of the substrate nitrogen was released as nitrate into the medium, and 61% of the carbon from uniformly 14C-labelled DNOC was recovered as 14CO2. The mixed microbial culture also decomposed 4-nitrophenol and 2,4-dinitrophenol but not 2,3-dinitrophenol, 2,6-dinitrophenol, 2,4-dinitrotoluene, 2,4 dinitrobenzoic acid or 2-sec-butyl-4,6-dinitrophenol (Dinoseb). Maximal degradation rates for DNOC by the bacterial biofilm immobilized on glass beads in fixed-bed column reactors were 30 mmol day-1 (1 reactor volume)-1, leaving an effluent concentration of less than 5 micrograms l-1 DNOC in the outflowing medium. The apparent Ks value of the immobilized mixed culture for DNOC was 17 microM. Degradation was inhibited at DNOC concentrations above 30 microM and it ceased at 340 microM, possibly because of the uncoupling action of the nitroaromatic compound on the cellular energy-transducing mechanism. PMID- 9390452 TI - The effect of nisin concentration and nutrient depletion on nisin production of Lactococcus lactis. AB - The kinetics of nisin production was studied in batch cultures using a construct of Lactococcus lactis subsp. lactis C2SmPrt-, containing a transposon (TnNip) that encodes nisin production. The introduction of TnNip into C2SmPrt- significantly lowered the specific growth rate and the maximum A620 reached was reduced from 15.2 to 11.0. The effect of nisin concentration and nutrient depletion on nisin production of the construct, C2SmPrt-(TnNip), was examined. Nisin production was found to be inhibited by high concentrations of nisin, when grown in excess nutrient, even though growth of the culture continued because nutrient limitation was not operating. However, in low nutrient concentrations nisin production was limited by nutrient depletion. The specific growth rate of C2SmPrt-(TnNip) was altered, by using different nutrient concentrations and different sugars, in order to examine the relationship between nisin production and growth. Nisin production was shown to be growth-associated for most of growth, but near the end of growth, when the specific growth rate was 0.05 h-1 or less, the production ceased. PMID- 9390453 TI - Cytochrome c peroxidase from a methylotrophic yeast: physiological role and isolation. AB - Mutant strains of the methylotrophic yeast Hansenula polymorpha defective in catalase (cat) and in glucose repression of alcohol oxidase synthesis (gcr1) have been isolated following multiple UV mutagenesis steps. One representative gcr1 cat mutant C-105 grows during batch cultivation in a glucose/methanol medium. However, growth is preceded by a prolonged lag period. C-105 and other gcr1 cat mutants do not grow on methanol medium without an alternative carbon source. A large collection of second-site suppressor catalase-defective (scd) revertants were isolated with restored ability for methylotrophic growth (Mth+) in the absence of catalase activity. These Mth+ gcr1 cat scd strains utilize methanol as a sole source of carbon and energy, although biomass yields are reduced relative to the wild-type strain. In contrast to the parental C-105 strain, H2O2 does not accumulate in the methanol medium of the revertants. We show that restoration of methylotrophic growth in the suppressor strains is strongly correlated with increased levels of the alternative H2O2-destroying enzyme, cytochrome c peroxidase. Cytochrome c peroxidase from cell-free extracts of one of the scd revertants has been purified to homogeneity and crystallized. PMID- 9390454 TI - Characterization of Leuconostoc mesenteroides NRRL B-512F dextransucrase (DSRS) and identification of amino-acid residues playing a key role in enzyme activity. AB - Dextransucrase (DSRS) from Leuconostoc mesenteroides NRRL B-512F is a glucosyltransferase that catalyzes the synthesis of soluble dextran from sucrose or oligosaccharides when acceptor molecules, like maltose, are present. The L. mesenteroides NRRL B-512F dextransucrase-encoding gene (dsrS) was amplified by the polymerase chain reaction and cloned in an overexpression plasmid. The characteristics of DSRS were found to be similar to the characteristics of the extracellular dextransucrase produced by L. mesenteroides NRRL B-512F. The enzyme also exhibited a high homology with other glucosyltransferases. In order to identify critical amino acid residues, the DSRS sequence was aligned with glucosyltransferase sequences and four amino acid residues were selected for site directed mutagenesis experiments: aspartic acid 511, aspartic acid 513, aspartic acid 551 and histidine 661. Asp-511, Asp-513 and Asp-551 were independently replaced with asparagine and His-661 with arginine. Mutation at Asp-511 and Asp 551 completely suppressed dextran and oligosaccharide synthesis activities, showing that at least two carboxyl groups (Asp-511 and Asp-551) are essential for the catalysis process. However, glucan-binding properties were retained, showing that DSRS has a two-domain structure like other glucosyltransferases. Mutations at Asp-513 and His-661 resulted in greatly reduced dextransucrase activity. According to amino acid sequence alignments of glucosyltransferases, alpha amylases or cyclodextrin glucanotransferases, His-661 may have a hydrogen-bonding function. PMID- 9390455 TI - Possible roles for a non-modular, thermostable and proteinase-resistant cellulase from the mesophilic aerobic soil bacterium Cellvibrio mixtus. AB - The widespread presence of cellulose-binding domains in cellulases from aerobic bacteria and fungi suggests the existence of a strong selective pressure for the retention of these non-catalytic modules. The complete nucleotide sequence of the cellulase gene, celA, from the aerobic soil bacterium Cellvibrio mixtus, was determined. It revealed an open reading frame of 1089 bp that encoded a polypeptide, defined as cellulase A (CelA), of M(r) 41,548. CelA displayed features characteristic of an endo-beta-1,4-glucanase, rapidly decreasing the viscosity of the substrate while releasing only moderate amounts of reducing sugar. Deletion studies in celA revealed that removal of 78 nucleotides from the 5' end or 75 from the 3' end of the gene led to the complete loss of cellulase activity of the encoded polypeptides. The deduced primary structure of CelA revealed an N-terminal signal peptide followed by a region that exhibited significant identity with the catalytic domains of cellulases belonging to glycosyl hydrolase family 5. These data suggest that CelA is a single-domain endoglucanase with no distinct non-catalytic cellulose-binding domain. Analysis of the biochemical properties of CelA revealed that the enzyme hydrolyses a range of soluble cellulosic substrates, but was inactive against Avicel, xylan or any other hemicellulose. CelA was resistant to proteolytic inactivation by pancreatic proteinases and surprisingly, in view of its mesophylic origin, was shown to be thermostable. The significance of these findings in relation to the role of single-domain cellulases in plant cell wall hydrolysis by aerobic microorganisms is discussed. PMID- 9390456 TI - Biosynthetic production of type II fish antifreeze protein: fermentation by Pichia pastoris. AB - Sea raven type II antifreeze protein (SRAFP) is one of three different fish antifreeze proteins isolated to date. These proteins are known to bind to the surface of ice and inhibit its growth. To solve the three-dimensional structure of SRAFP, study its ice-binding mechanism, and as a basis for engineering these molecules, an efficient system for its biosynthetic production was developed. Several different expression systems have been tested including baculovirus, Escherichia coli and yeast. The latter, using the methylotrophic organism Pichia pastoris as the host, was the most productive. In shake-flask cultures the levels of SRAFP secreted from Pichia were up to 5 mg/l. The recombinant protein has an identical activity to SRAFP from sea raven serum. In order to increase yields further, four different strategies were tested in 10-l fermentation vessels, including: (1) optimization of pH and dissolved oxygen, (2) mixed feeding of methanol and glycerol with Mut(s) clones, (3) supplementation of amino acid building blocks, and (4) methanol feeding with Mut+ clones. The mixed feeding/Mut(s) strategy proved to be the most efficient with SRAFP yields reaching 30 mg/l. PMID- 9390457 TI - Efficient production of a functional mouse/human chimeric Fab' against human urokinase-type plasminogen activator by Bacillus brevis. AB - Expression/secretion vectors for the production of Fab' and single-chain (sc) Fab' by Bacillus brevis have been constructed. For the production of Fab', the cDNAs encoding the L chain and Fd' fragment (Fd with the hinge region) of a mouse human chimeric Fab' against human urokinase-type plasminogen activator were fused directly with the translation-start and signal-peptide-encoding regions of the mwp gene, the gene for one of the major cell-wall proteins of Bacillus brevis. The two fused genes were placed tandemly downstream from the promoter of the cell wall protein gene operon (cwp) of B. brevis. For the production of scFab', the two cDNAs were linked with a synthetic oligonucleotide encoding a flexible peptide linker of 17 or 24 amino acids, and fused with the translation start and signal-peptide-encoding regions of the mwp gene. Fab' was efficiently produced by B. brevis, being accumulated at a level of 100 mg/l in the culture medium in a simple shake-flask culture, which is the highest level obtained so far for a gram positive bacterium. On the other hand, the scFab' remained at a level of a few milligrams per liter in the culture medium. The Fab' produced by B. brevis showed comparable antigen-binding activity to that of the parental antibody. The L chain and Fd' fragment, constituting the Fab', had the correct N-terminal amino acid sequences. These results indicate that B. brevis is a very promising host for the production of native Ig fragments. PMID- 9390458 TI - Competition of plasmid-bearing Pseudomonas putida strains catabolizing naphthalene via various pathways in chemostat culture. AB - Plasmid-carrying Pseudomonas putida strains degrade naphthalene through different biochemical pathways. The influence of various combinations of host bacteria and plasmids on growth characteristics and competitiveness of P. putida strains was studied in chemostat culture at a low dilution rate (D = 0.05 h-1) with naphthalene as the sole source of carbon and energy. Under naphthalene limitation, the plasmid-bearing strains degrading naphthalene that use catechol 1,2-dioxygenase for catechol oxidation (ortho pathway), were the most competitive. The strains bearing plasmids that control naphthalene catabolism via catechol 2,3-dioxygenase (meta pathway), were less competitive. Under these conditions the strain carrying plasmid pBS4, which encodes for naphthalene catabolism via gentisic acid, was the least competitive. PMID- 9390459 TI - Genetic immobilization of cellulase on the cell surface of Saccharomyces cerevisiae. AB - We tried genetically to immobilize cellulase protein on the cell surface of the yeast Saccharomyces cerevisiae in its active form. A cDNA encoding FI carboxymethylcellulase (CMCase) of the fungus Aspergillus aculeatus, with its secretion signal peptide, was fused with the gene encoding the C-terminal half (320 amino acid residues from the C terminus) of yeast alpha-agglutinin a protein involved in mating and covalently anchored to the cell wall. The plasmid constructed containing this fusion gene was introduced into S. cerevisiae and expressed under the control of the glyceraldehyde-3-phosphate dehydrogenase promoter from S. cerevisiae. The CMCase activity was detected in the cell pellet fraction. The CMCase protein was solubilized from the cell wall fraction by glucanase treatment but not by sodium dodecyl sulphate treatment, indicating the covalent binding of the fusion protein to the cell wall. The appearance of the fused protein on the cell surface was further confirmed by immunofluorescence microscopy and immunoelectron microscopy. These results proved that the CMCase was anchored on the cell wall in its active form. PMID- 9390460 TI - Effect of growth rate on alpha-amylase production by Streptomyces sp. IMD 2679. AB - The alpha-amylase of Streptomyces sp. IMD 2679 was subject to catabolite repression. Four different growth rates were achieved when the organism was grown at 40 degrees C and 55 degrees C in the presence and absence of cobalt, with an inverse relationship between alpha-amylase production and growth rate. Highest alpha-amylase yields (520 units/ml) were obtained at the lowest growth rate (0.062 h-1), at 40 degrees C in the absence of cobalt, while at the highest growth rate (0.35 h-1), at 55 degrees C in the presence of cobalt, alpha-amylase production was decreased to 150 units/ml. As growth rate increased, the rate of specific utilisation of the carbon source maltose also increased, from 46 to 123 micrograms maltose (mg biomass)-1 h-1. The pattern and levels of alpha glucosidase (the enzyme degrading maltose) detected intracellularly in each case, indicate that growth rate effectively controls the rate of feeding of glucose to the cell, and thus catabolite repression. PMID- 9390461 TI - Influence of phosphate on rhamnose-containing exopolysaccharide rheology and production by Klebsiella I-714. AB - Physiological conditions enhancing rhamnose-containing polysaccharide synthesis by Klebsiella I-714 were studied in batch culture (0.3-l and 2-l bioreactors). The four carbon sources tested, sucrose, sorbitol, Neosorb and Cerelose, allowed exopolysaccharide production. Larger amounts of polymer were produced when high carbon/nitrogen ratios and complex nitrogen sources were used. Exopolysaccharide synthesis was greatest at 30 degrees C, which was a suboptimal growth temperature. A reduction in the phosphate content of the medium enhanced rhamnose containing polysaccharide production. When the initial carbon source concentration was augmented, byproducts other than exopolysaccharide were formed. Rhamnose-containing polysaccharide rheology can be modulated by changing the phosphate content of the medium. PMID- 9390462 TI - Relationship between cadmium sensitivity and degree of plasma membrane fatty acid unsaturation in Saccharomyces cerevisiae. AB - The sensitivity of Saccharomyces cerevisiae to the redox-active metal copper has recently been found to be influenced by cellular fatty acid composition. This study sought to investigate whether fatty acid composition affected plasma membrane permeabilisation and whole-cell toxicity induced by the redox-inactive metal cadmium. S. cerevisiae NCYC 1383 was enriched with the polyunsaturated fatty acids linoleate (18:2) and linolenate (18:3) by growth in 18:2- or 18:3 supplemented medium. Incorporation of the exogenous fatty acids resulted in them comprising more than 65% of the total fatty acids in plasma membrane lipids. Inhibition of cell division in the presence of Cd(NO3)2 was accentuated by growth in the presence of a polyunsaturated fatty acid. Furthermore, susceptibility to Cd(2+)-induced plasma membrane permeabilisation increased with the degree of fatty acid unsaturation. Thus, during exposure to Cd2+, K+ efflux from 18:2- and 18:3-enriched cells was up to 2.5-fold or 3-fold greater, respectively than that from unsupplemented cells. In addition, reductions in cell viability during exposure to Cd2+ were most marked in polyunsaturated-fatty-acid-supplemented cells. At certain times, unsupplemented Cd(2+)-exposed cells displayed up to 7 fold greater viability than supplemented Cd(2+)-exposed cells. The study demonstrates that the toxicity of the redox-inactive metal Cd2+ towards S. cerevisiae becomes markedly amplified with increased cellular and plasma membrane fatty acid unsaturation. PMID- 9390463 TI - The ability of soil-borne fungi to degrade organophosphonate carbon-to-phosphorus bonds. AB - The ability of a wide variety of soil-borne fungal strains to degrade four structurally different compounds containing P-C bonds, namely the naturally occurring amino acid ciliatine, the popular herbicide glyphosate, phosphonoacetic acid and 2-amino-3-phosphonopropionic acid, was studied in order to show that soil fungi may play an important role in the biodegradation of organophosphonates. Most of the strains appeared to utilize ciliatine as the sole source of phosphorus for growth. Only a limited number of strains were able to grow on the other phosphonates used in this work. The strains of Trichoderma harzianum, Scopulariopsis sp. and Aspergillus niger chosen for more detailed study show the ability to degrade ciliatine, glyphosate and also amino(3 methoxyphenyl)methylphosphonic acid effectively. PMID- 9390464 TI - [The 33rd autumn meeting of the Japan Radiological Society. Nara City, Japan. October 7-9, 1997. Abstracts]. PMID- 9390465 TI - [The 98th Congress of the Japanese Society of Otolaryngology. Ohsaka, Japan. May 22-24, 1997. Abstracts]. PMID- 9390466 TI - The Institute of Medical Illustrators and Glasgow Caledonian University BSc degree in Medical Illustration. AB - This paper relates the history leading to the validation of the Institute of Medical Illustrators (IMI) Diploma as a part-time, work-based, Bachelor of Science Degree by Glasgow Caledonian University (GCU) on the 11th June 1996. It also outlines a description of the structure and content of this degree programme. The BSc is a joint venture between GCU, who are the awarding academic institution, and the IMI, who are the professional examining body. The Institute hopes that it will become a standard qualification for the profession. PMID- 9390467 TI - The Institute of Medical Illustrators and Glasgow Caledonian University BSc in Medical Illustration assessment system. AB - As a part-time work-based degree with students registered from all parts of the United Kingdom, and potentially from abroad, the Bachelor of Science (BSc) in Medical illustration does not have assessment in the conventional form as used by full-time courses undertaken in a university. It incorporates the expertise of medical illustrators actually working in the profession. The unique nature of the Institute of Medical Illustrators (IMI) and Glasgow Caledonian University (GCU) BSc has required the development of an assessment system that satisfies the quality assurance demands of both the awarding and professional body. This paper details the system devised to meet the needs of the provision and to ensure consistent assessment. PMID- 9390468 TI - 'Conversion course' to allow holders of the IMI Diploma in Medical Illustration to gain a BSc in Medical Illustration from Glasgow Caledonian University. AB - The 'conversion course' described in this paper has been set up following discussions between the Institute of Medical Illustrators (IMI) and Glasgow Caledonian University (GCU). The 'conversion course' will take the form of a degree triple module with a credit rating of 60 Scottish Credit and Accumulation Transfer (SCOTCAT) credits at Scottish Degree (SD) level 3. This module will require the student to undertake an extended theoretical based investigative project. The project will permit the student to study in-depth an aspect of his/her specialist interest that has a particular professional relevance. The topic of the project will be negotiated between the student and a scrutiny panel under the aegis of the department of Biological Sciences at Glasgow Caledonian University. The project will be written up in the style of an academic paper for the Institute's journal. Successful students will be awarded the BSc in Medical Illustration. PMID- 9390469 TI - Using accreditation of prior learning and accreditation of prior experiential learning for entry on to the BSc in Medical Illustration. AB - The Accreditation of Prior Learning (APL) and Accreditation of Prior Experiential Learning (APEL) scheme described in this paper has been prepared following discussions between the Institute of Medical Illustrators (IMI) and Glasgow Caledonian University (GCU), departments of Learning and Educational Development and Biological Sciences. The scheme gives specific academic credit under the Scottish Credit Accumulation and Transfer (SCOTCAT) scheme for learning gained from experience, allowing access onto the Bachelor of Science Degree (BSc) in Medical Illustration to potential students who do not have the required Higher National Diploma (HND) or equivalent entry qualification. The need, rationale and structure of the scheme are described. PMID- 9390470 TI - Research for medical illustrators. Part 2: Trial design, protocols, results and reporting. AB - Medical illustrators have long been involved in the research projects of other workers, by providing photographic and other visual records. These illustrations may be used for measurement purposes, in which case it is important that medical illustrators understand the requirements for using tested protocols, which ensure that all variables of the recording medium are controlled. Now that there are degree courses in medical illustration an increasing number of medical illustrators in the UK are instigating their own research projects, making it all the more important that they understand the research process. In this second of a two-part article, consideration is given to: the design of experiments and research trials; the production of research protocols; data analysis and interpretation; the writing of the research report or dissertation; and the role of the supervisor or mentor. PMID- 9390471 TI - Thomas Albert Longmore, Hon FSR, FRPS (1905-1957). PMID- 9390472 TI - From chromatograph to cuspid: dental clinical research and the future of technology transfer. PMID- 9390473 TI - Melvin L. Moss and the functional matrix. PMID- 9390474 TI - Structural and functional association between substance P- and calcitonin gene related peptide-immunoreactive nerves and accessory cells in the rat dental pulp. AB - Defense mechanisms of the dentin/pulp complex involve a variety of biological systems in which immunocompetent cells, the nervous system, and the vascular supply play important roles. In the present study, pulpal accessory cells were examined regarding (i) their structural relationship to nerves and (ii) how the functional capacities of these cells were affected by neuropeptides. Micro anatomic association was investigated in the normal rat molar pulp with the use of double-immunofluorescence staining and dual-channel confocal laser scanning microscopy. Examinations of confocal laser scanning microscopic images from single focal planes revealed the presence of apparent contacts between thin, varicose nerve fibers and immunocompetent cells, indicating proximity between these two structures. The close associations were most frequently observed in the para-odontoblastic region of the coronal pulp, where more than 70% of class II antigen-expressing (OX6+) cells showed proximity to nerve fibers immunoreactive to calcitonin gene-related peptide. The corresponding figure for substance P was about 50%. ED2+ macrophages closely associated with nerves were less frequently observed. Functional studies conducted in vitro demonstrated that 10(-9) to 10( 7) mol/L of substance P significantly increased (p < 0.05), while 10(-7) to 10( 6) mol/L of calcitonin gene-related peptide suppressed (p < 0.01) proliferation of purified T-lymphocytes stimulated with sub-optimal concentrations of concanavalin A in the presence of rat incisor pulpal cells as accessory cells. These data suggest that pulpal sensory nerve fibers and their products may have an influence upon the immune defense of the dental pulp. PMID- 9390475 TI - HLA-D types and serum IgG responses to Capnocytophaga in diabetes and periodontitis. AB - Serum IgG responses to the cell envelope proteins (CEPs) from Capnocytophaga sputigena, Capnocytophaga ochracea, and Capnocytophaga gingivalis were examined in periodontally healthy and periodontitis subjects, both with and without type 1 diabetes (n = 60). Serum IgG responses to CEPs were determined by immunoblotting with biotin-goat anti-human IgG and an alkaline phosphatase-streptavidin system. Reactivity was analyzed by transmission densitometry, digitization, and computer manipulation. The patients with diabetes showed significantly (p < 0.01) fewer responses to 14 CEPs (from 81 to 10 kDa) from C. sputigena, 5 CEPs (from 90 to 17 kDa) from C. gingivalis, and the 27-kDa CEP from C. ochracea than in the non diabetic group. The periodontitis patients showed significantly (p < 0.01) fewer responses to the 25- and 11-kDa CEPs from C. sputigena, the 125- and 17-kDa CEPs from C. gingivalis, and the 42-kDa CEP from C. ochracea than in the periodontally healthy group. HLA-DR4, HLA-DR53, and HLA-DQw3 were associated with periodontitis, while only HLA-DR4 was associated with diabetes (p < 0.02). Significant (p < 0.01) correlations were found between HLA-DR2 and IgG reactivity patterns associated with non-diabetics, and between HLA-DR4 and IgG reactivity patterns associated with diabetic and periodontitis subjects. These results indicate that both type 1 diabetics and periodontitis subjects have a depressed IgG antibody profile to Capnocytophaga, which may account for an increased susceptibility to periodontitis infection. Periodontitis in type 1 diabetes may be related more to the HLA-D type and altered immune function than to the diabetes itself. PMID- 9390476 TI - Profile of cytokine mRNA expression in chronic adult periodontitis. AB - Chronic inflammation induced by bacteria often leads to host-mediated destruction of tissues adjacent to the sites of microbial insult. The chronic inflammatory process of adult periodontitis results in the destruction of supporting osseous and connective tissues of the teeth. We hypothesized that virulence factors of periodontal pathogens such as lipopolysaccharide stimulate inflammatory cytokine expression by mononuclear cells of the host which contribute to disease development. In this study, to elucidate the role of these cytokines in chronic adult periodontitis, we tested whether the prevalence of mRNA for inflammatory cytokines generally associated with mononuclear phagocytes was higher in diseased than in healthy gingival tissue. Gingival mononuclear cells or whole gingival biopsies from 32 adult periodontitis patients and five healthy individuals used as controls were evaluated for inflammatory cytokine mRNA expression by reverse transcription polymerase chain-reaction (RT-PCR) procedures. The cytokines assessed included IL-1 alpha, IL-1 beta, IL-1ra, IL-6, IL-8, IL-12, IL-13, TNF alpha, TGF-beta, and IFN-gamma. The monocyte/macrophage lipopolysaccharide (LPS) receptor CD14 was also assessed. Results showed that TNF-alpha mRNA was present significantly more frequently in diseased than in healthy biopsies, whereas IL-1 alpha, IL-1 beta, and IL-1ra mRNA were found in most (from 80 to 100%) healthy tissues. Message for CD14 was present in both healthy and diseased tissue samples (100%). This study provides evidence for a major role of TNF-alpha in chronic adult periodontitis. Moreover, our results suggest that the mononuclear cells derived from periodontal tissues have the capacity to respond to components of periodontal pathogens and express both pro- and anti-inflammatory cytokines in these tissues. PMID- 9390477 TI - Prevalence and distribution of six capsular serotypes of Porphyromonas gingivalis in periodontitis patients. AB - Previous reports have described six serotypes based on K antigens in Porphyromonas gingivalis strains. The purpose of the present study was to investigate the prevalence and distribution of these serotypes in 185 patients with P. gingivalis-associated periodontitis. Polyclonal rabbit antisera, raised against each of the different type strains, were used in double-immunodiffusion and immunoelectrophoresis assays. In addition, a subset of 76 strains was investigated for the presence of capsular structures by means of the India ink and Bruce White staining techniques. These strains were also tested for auto aggregation in phosphate-buffered saline (PBS). All six K serotypes were present in the study sample. In total, 84 (45.4%) patients were colonized with a K typeable P. gingivalis strain with a predominance of types K5 (12%) and K6 (23.2%). A correlation was found between arbitrary age categories and the prevalence of currently known K serotypes, which were found in 60% of patients aged 12 to 30 years, in 49% of patients aged 31 to 50, and in 25% of patients aged 51 to 70 years. In the subset of 76 P. gingivalis strains, 32 (42.1%) were K typeable. Fifty-three strains (69.7%) showed microscopic evidence of encapsulation, suggesting the existence of K serotypes other than K1 to K6. Twenty-one strains (27.6%) auto-aggregated in PBS and were not K-typeable, nor did they show any evidence of encapsulation. It was concluded that the majority of clinical P. gingivalis isolates is encapsulated and that encapsulation is associated with the presence of a K antigen. Auto-aggregation seems to be associated with the absence of a capsular structure and, consequently, the absence of a K antigen. PMID- 9390478 TI - Arrest of root surface caries in situ. AB - This study tests the hypothesis that daily oral hygiene combined with topical fluoride arrests active root-surface caries lesions without changing the mineral content of the lesions. Therefore, changes in mineral content and distribution were studied in root surfaces during caries lesion development and subsequent arrest of lesion progression in situ. In 18 subjects, lesions were developed during 3 months in sound root-surface specimens inserted into lower partial dentures. After 3 months, ground sections were prepared from each lesion prior to re-insertion of the specimens into the dentures. In addition, one sound root specimen was added per subject. During the following 3 months, half of the subjects cleaned both sound and carious specimens once a day with an 1100-ppm fluoride toothpaste, and the specimens were treated twice with 2% NaF for 2 min in situ. The other half of the subjects continued the experiment without cleaning. During the initial three-month period, all specimens developed subsurface lesions extending 187 to 583 microm into the dentin. Lesion depth increased somewhat in both experimental groups during the following 3 months (P > or = 0.1). There was a non-significant increase in mineral loss in the plaque covered specimens (P = 0.08). However, the total mineral content of specimens subjected to plaque removal and topical fluoride did not change. This treatment resulted in an increased mineral content in the surface layer (P < 0.01) and formation of a zone of higher mineral content within the body of the lesion. The sound root surfaces which had been cleaned for a three-month period showed mineral uptake in the surface layer, occasionally associated with subsurface demineralization extending 20 to 70 microm into the tissue. The mineral loss of these specimens was significantly smaller than that of plaque-covered surfaces (P < 0.001). It is concluded that daily plaque removal and topical fluoride use influence the distribution of mineral in sound and carious root surfaces and may arrest lesion progression without affecting the total mineral content. PMID- 9390479 TI - Prevalence and depth of artificial caries-like lesions adjacent to cavities prepared in roots and restored with a glass ionomer or a dentin-bonded composite material. AB - One potential advantage of glass-ionomer materials for the treatment of root caries is their ability to release fluoride and so resist cariogenic attack. A commercially available composite material has also been reported to release fluoride which reduced caries lesions in the tooth tissue adjacent to it. The aim of this study was to assess the effectiveness of a conventional glass-ionomer restoration compared with a dentin-bonded, fluoride-releasing, composite restoration when exposed to a microbial artificial caries system. Artificial caries-like lesions produced in relation to the restorations were examined and classified either as outer (surface) lesions or as wall lesions. A split-unit experimental design allowed for within-tooth comparisons of the 2 experimental restorations at different sites on the root surface. These were either totally within the root surface or positioned at the amelo-cemental junction. Outer lesion depths were significantly (p < 0.001) shallower at all sites adjacent to the glass ionomer when compared with the composite restorations. Wall lesions were significantly (p < 0.01) more prevalent adjacent to the composite material. In addition, the cavity margin position significantly (p < 0.05) affected the incidence of wall lesions, particularly in the composite group. In conclusion, glass ionomer was successful in reducing the caries-like lesion production in the adjacent root surface. This resulted from improved marginal integrity and fluoride release from this material when compared with the composite bonding system used. PMID- 9390480 TI - The relationship between clinical tooth status and receipt of sealants among child Medicaid recipients. AB - This study investigated the association between caries status and sealant need at a prior survey and subsequent sealant use in a Medicaid program. Clinical data from a 1986-87 statewide epidemiological survey (N = 8026) representative of North Carolina (NC) schoolchildren (grades K-12) were linked with all NC Medicaid dental claims submitted during 1987-92, yielding 570 children in the survey who had at least one dental visit during 1987-1992. From the 570, 390 children were included: 71 who received sealants (S) and 319 who received non-sealant care (NS). Children were excluded based on age, having preexisting sealants, or having no sealant-eligible molars or premolars. S and NS were compared on baseline dfs, DMFS, and sealant need, controlling for the patient's age, number of visits, and the provider's propensity to seal. At all ages, NS was twice as likely to have had prior dfs or DMFS (OR = 2.04, 95% CI = 1.15, 3.70). The association between sealant receipt and prior sealant need varied by age. At 6 to 11 years, S and NS had equal likelihood of sealant need (OR = 1.41, 95% CI = 0.62, 3.18). At 12 to 15 years, NS had a greater likelihood of sealant need (OR = 6.82, 95% CI = 1.60, 29.08). Caries-free status was associated with subsequent sealant receipt. Prior sealant need caused variability in dentists' decisions, depending on the child's age and past caries experience. Sealants were used infrequently by most providers and for a minority of patients. These findings are important for the Medicaid program and for future non-randomized studies of sealant effectiveness. PMID- 9390481 TI - Connections between polarization curves and log(a[i]/a[ref])-pe diagram. AB - When reading papers concerning studies of corrosion of dental amalgam and its phases by means of polarization curves, one often finds it difficult to understand the reasons for the chemical reactions proposed from the form of the polarization curve. Thermodynamic data represented in the form of log(a[i]/a[ref])-pe diagrams, i.e., the logarithm of the activity of a metal or an alloy with reference to the activity of the corresponding metal ion, as a function of pe (a recalculated form of the potential), make it possible for one to determine which chemical reactions can occur on the specimen surface or in the solution within the potential difference used in the polarization experiment and to decide which of these reactions is the most probable. The hypothesis examined in this study is that a log(a[i]/a[ref])-pe diagram can be used in the interpretation of polarization curves. Potentiodynamic polarization curves and log(a[i]/a[ref])-pe diagrams were compared for the corrosion of Ag, Hg, and gamma 1 with and without Sn. It was found that there was a connection between the polarization curves and the log(a[i]/a[ref])-pe diagrams. From the composition of the specimens and the solution and by means of thermodynamic data, pe values for solid corrosion products and relative concentrations of soluble complexes at these pe values were obtained independently of the polarization curves. A much more reliable value for the nobility of metals and alloys was attained by use of the log(a[i]/a[ref])-pe diagrams than by use of the potential of the starting point of the polarization curves. It was found that pe corresponding to the potential of the starting point of the polarization curves in de-aerated synthetic saliva was obtained about two pe units before pe of the most insoluble solid compound formed on the specimen surface or in the solution. PMID- 9390482 TI - Paint the ceiling: reflections on illness. PMID- 9390483 TI - Multicenter, randomized, prospective trial of early tracheostomy. AB - OBJECTIVES: Determine the effect of early (days 3-5) or late (days 10-14) tracheostomy on intensive care unit length of stay (ICU LOS), frequency of pneumonia, and mortality, and evidence of short-term or long-term pharyngeal, laryngeal, or tracheal injury in head trauma, non-head trauma, and critically ill nontrauma patients. STUDY DESIGN: Randomized, prospective. SETTING: Five Level I trauma centers. METHODS: Data were obtained prospectively and included Acute Physiology and Chronic Health Evaluation III score (AIII), Glasgow Coma Scale score, Emergency Room Trauma Score, Injury Severity Score, Acute Injury Score, type of endotracheal tube or tracheostomy, level of positive end-expiratory pressure, and peak inspiratory pressure. Patients were to undergo laryngoscopy for detection of injury according to the Lindholm criteria at the time of endotracheal tube or tracheostomy removal and be reevaluated at 3 to 5 months after discharge. RESULTS: One hundred fifty-seven patients were entered, 127 to early randomization (3-5 days) and 28 to late randomization (10-14 days); however, only 112 patients with early and 14 with late randomization had completed data forms for the primary study goals. An additional 22 patients from the early entry groups were rerandomized late. Early randomization data: the AIII score was higher (p < 0.05) in the head trauma tracheostomy (65 +/- 4) than in the nontracheostomy group (51 +/- 4) and in the nontrauma tracheostomy (92 +/- 6) than in the nontracheostomy group (68 +/- 7), but was equivalent in the non-head trauma group. Glasgow Coma Scale score, Emergency Room Trauma Score, Injury Severity Score, Acute Injury Score, positive end-expiratory pressure, and peak inspiratory pressure were not significantly different in any of the groups. There were no significant differences in ICU LOS, frequency of pneumonia, or death in any of the groups after either early or late tracheostomy compared with continued endotracheal intubation. Only 83 patients underwent postextubation laryngoscopy. There were no significant differences between the groups; however, there were trends to more vocal cord ulceration and subglottic inflammation in the continued intubation group. No patient was seen in this study with late vocal cord or laryngeal stenosis; there were no tracheal-innominate artery fistulae. Seven of the patients with abnormal findings at extubation had normal 3- to 5-month postextubation laryngoscopy. CONCLUSION: Physician bias limited patient entry into the study. Although there were higher AIII scores in the head trauma early tracheostomy patients, there were no differences in the primary end points of ICU LOS, pneumonia, or death in any of the groups studied. Long-term endoscopic follow-up was poor, but no known late tracheal stenosis was seen. PMID- 9390484 TI - Nonoperative management of splenic injury: are follow-up computed tomographic scans of any value? AB - OBJECTIVE: To determine the value of follow-up abdominal computed tomography in patients with splenic trauma managed nonoperatively. DESIGN: Retrospective chart review. MATERIALS AND METHODS: A total of 108 consecutive patients with splenic injuries treated at a single institution from 1990 to 1996 were studied. All admission and follow-up computed tomographic (CT) scans were reviewed by the authors. RESULTS: Initial management was surgical in 35 patients (32%) and intentionally nonoperative in 73 patients (68%). Nonoperative management was successful in 45 of 49 adults (92%) and 21 of 24 children(88%). Sixty-two follow up abdominal CT scans were obtained in 49 patients. Information that affected management was evident on only one follow-up CT scan performed in the absence of clinical indications. Potential savings in hospital and physician charges for routine follow-up CT scans in this study were $54,302.00. CONCLUSIONS: Follow-up abdominal CT scans are not routinely necessary in patients with splenic injuries managed nonoperatively. PMID- 9390485 TI - Cost-effective method for bedside insertion of vena caval filters in trauma patients. AB - BACKGROUND: The need for patient transport for inferior vena cava (IVC) filter placement impacts patient safety, comfort, charges, and nursing care. Bedside, ultrasound-guided IVC filter placement may offer an acceptable, cost-effective alternative. METHODS: Prospective cohort study of 55 consecutive trauma patients requiring IVC filter placement. During a 13-month period (August of 1995 September of 1996), patients meeting criteria for IVC filter were evaluated. Complications were recorded, and the potential financial savings were determined. RESULTS: Of 3,172 trauma admissions, 55 patients met IVC filter criteria and 49 patients had IVC filters placed under ultrasound guidance. In six patients (10.9%), ultrasound guided filter placement failed. There were four complications in four patients (8.2%). Over 13 months, charges were reduced by $69,800 when compared with radiology suite placement and $118,300 when compared with operative placement. CONCLUSIONS: Ultrasound guided, bedside placement of IVC filters is a safe, cost-effective method of pulmonary embolism prophylaxis in select trauma patients. PMID- 9390486 TI - Secretory immunoglobulin A blocks hypoxia-augmented bacterial passage across Madin-Darby canine kidney cell monolayers. AB - OBJECTIVE: To study the relative impact of previous hypoxic exposure and the addition of secretory immunoglobulin A (IgA) on bacterial translocation. DESIGN: In vitro randomized experimental study. MATERIALS AND METHODS: Transfected Madin Darby canine kidney epithelial cells were grown as monolayers in a two-chamber tissue culture system. Stationary growth phase Escherichia coli M14 were inoculated in the apical chamber with medium or medium containing polymeric secretory IgA. Tissue culture dishes were then placed in a 21 or 5% O2 incubator environment for 90 minutes followed by a 21% O2 environment. Medium from the basal compartment was then obtained at timed intervals for bacterial culture. MEASUREMENT AND MAIN RESULTS: Bacterial translocation increased with time in co culture. Previous hypoxic exposure augmented translocation across the monolayers. The addition of IgA blocked translocation under both normoxic and hypoxic conditions. CONCLUSION: Secretory IgA is important in mucosal defense under both normal and shock conditions. PMID- 9390487 TI - Posttraumatic empyema thoracis: a 24-year experience at a major trauma center. AB - The purpose of this paper is to review the outcome of patients with posttraumatic empyema thoracis. Between April 1972 and March 1996, the Division of Cardiothoracic Surgery at the King-Drew Medical Center managed or was consulted on 5,474 trauma patients (4,584 patients with penetrating injuries and 890 with blunt injuries) who were admitted emergently for thoracic and thoracoabdominal injuries and who underwent tube thoracostomy. Patients were not given routine prophylactic antibiotics merely because they had a chest tube placed. Based on our previous reports on thoracic trauma, our criteria for empiric antibiotic administration included (1) emergent or urgent thoracotomy, (2) soft-tissue destruction of the chest wall by shotgun injuries, (3) lung contusion with hemoptysis, (4) associated abdominal trauma requiring exploratory laparotomy, or (5) associated open long-bone fractures. Eighty-seven of these 5,474 patients developed posttraumatic empyema thoracis, for an incidence of 1.6%. These 87 patients were treated with tube thoracostomy, image-guided catheter drainage, or open thoracotomy with decortication. Seventy-nine of 87 patients (91%) were cured without conversion to open thoracostomy. Four patients required conversion to open thoracostomy, and there were three deaths. Even though a majority of our patients required decortication, successful management of posttraumatic empyema thoracis also was achieved with closed-tube thoracostomy or image-guided catheter drainage based on clinical and radiographic findings with appropriate patient selection. When thoracic empyema did occur in our group, Staphylococcus aureus was the most common microbe isolated, followed by anaerobic bacteria. In correlating microbiologic data with outcomes, S. aureus, especially methicillin resistant S. aureus, was the most frequent cause of antibiotic failure. Because of the low incidence of posttraumatic empyema thoracis, we do not recommend routine antibiotic prophylaxis for all trauma patients who undergo closed-tube thoracostomy. A review of the role of tube thoracostomy, intrapleural fibrinolytic therapy, image-guided catheter drainage, video-assisted thoracoscopy, and open thoracotomy for the management of thoracic empyema is provided. PMID- 9390488 TI - The importance of the command-physician in trauma resuscitation. AB - OBJECTIVE: Definitive trauma team leadership, although difficult to measure, has been shown to improve trauma resuscitation performance. The purpose of this study was to evaluate the effect of an identified command-physician on resuscitation performance. In addition, the leadership capability of four physician combinations functioning as command-physician was studied. DESIGN: Retrospective review. METHODS: Videotapes of trauma resuscitations performed at a Level I trauma center over a 25-month period were reviewed. The presence of an identified command-physician was determined by multidisciplinary consensus. Resuscitation performance was measured by compliance with three objective criteria: primary survey, secondary survey, and definitive plan; and two subjective criteria: orderliness, and adherence to Advanced Trauma Life Support protocol. Performance was then analyzed (1) as a function of the presence or absence of a command physician, and (2) between four identified physician combinations: AF (attending surgeon + trauma fellow); F (trauma fellow); ASR (attending surgeon + senior surgical resident); SR (senior surgical resident). Chi square and the Mann Whitney U tests were applied. RESULTS: A total of 425 trauma resuscitations were reviewed. A command-physician was identified (CP[Pos]) in 365 resuscitations (85.7%); no command-physician was identified (CP[NEG]) in 60 (14.3%). Compliance with completion of secondary survey (81.4%) and formulation of a definitive plan (89.6%) was significantly higher in the CP(POS) group. Subjective scores for orderliness and adherence to Advanced Trauma Life Support protocol were significantly higher in the CP(POS) group. In the CP(POS) resuscitations, formulation of a definitive plan was lower in SR when compared with the other three physician combinations. CONCLUSIONS: An identified command-physician enhances trauma resuscitation performance. Completion of the primary and secondary survey is not affected by the physician combination. Prompt formulation of a definitive plan is facilitated by the active involvement of an attending traumatologist or a properly mentored trauma fellow. PMID- 9390489 TI - Complications of plate fixation in fresh displaced midclavicular fractures. AB - BACKGROUND: The role of plate fixation in the management of fresh displaced midclavicular fractures is unsettled. The objective of this study was to evaluate the drawbacks and pitfalls of this treatment method. METHODS: We analyzed the complications encountered in 103 consecutive adult patients with severely displaced fresh fractures of the middle third of the clavicle who were treated by open reduction and internal fixation using AO/ASIF plates. These 103 patients accounted for 9.5% of the 1,081 patients with fresh midclavicular fractures seen between 1989 and 1995. The mean age of the 103 patients was 33.4 years (range, 19 62 years). RESULTS: Seventy-nine patients had an uneventful recovery, whereas 24 (23%) suffered one or several complications. The major complications included deep infection, plate breakage, nonunion, and refracture after plate removal. The most common of the minor complications was plate loosening resulting in malunion. The infection rate was 7.8%. A total of 14 reoperations were performed because of the complications. Permanent nonunion ensued in two patients. A severely comminuted fracture (relative risk of failure, 5.15) as well as a state of alcohol intoxication on admission (relative risk of failure, 3.12) were identified as markers of increased complication risk. CONCLUSIONS: Patient noncompliance with the postoperative regimen could be suspected to have been a major cause of the failures. The high complication rate supports a reserved attitude toward plate fixation of fresh midclavicular fractures. The method should be reserved for patients who have trustworthy personal motives for quick pain relief and functional recovery. PMID- 9390490 TI - Free fibula osteoseptocutaneous graft for reconstruction of segmental femoral shaft defects. AB - Seventeen major reconstructions of the femoral shaft using vascularized fibula osteoseptocutaneous grafts were performed from August 1984 to September 1993. Patients were 14 males and 3 females, with an average age of 34 years. All patients had sustained high-energy trauma in motor vehicle crashes and had bone defects averaging 10 cm. The skeletal defect was primary attributable to bone loss at the time of injury (2 cases) or secondary after infection and sequestrectomy (15 cases). Vascularized fibular transfer was performed at an average of 6 months after trauma. The fibular graft was inserted as a single strut in 10 cases and as a double-barrel composite in 7 cases. Patients were evaluated at an average of 43 months after surgery. All grafts eventually united, and no patient showed evidence of recurrent or persistent infection. The average time to radiologic union was 8 months, and the average time to full weight bearing was 14 months. Secondary bone grafting and internal fixation were required in five cases because of delayed union, stress fracture, or screw loosening. All cases of delayed union and stress fracture were in those reconstructed by single-strut fibular graft. Four cases (24%) required quadriceps plasty and arthrolysis. The final average arc of active knee motion was from 0 to 80 degrees. Limb length discrepancy ranged from 0 to 7 cm (average, 3 cm). Five cases (29%) had varus deformity averaging 30 degrees. The fibular graft hypertrophied to 100% of the femoral circumference in cases followed for 3 years. Donor site morbidity was negligible. At the time of final follow-up, 13 patients had returned to their original jobs, two were permanently disabled because of below-knee amputation, and two were retired. The study suggests that vascularized fibula osteoseptocutaneous transfer is a valuable procedure for reconstruction of large, previously infected femoral shaft defects. PMID- 9390491 TI - Staged management of infected humeral nonunion. AB - Fourteen patients with infected humeral nonunion complicated by sinus discharge were treated with a staged protocol consisting of (1) radical debridement with local antibiotic beads implantation, and (2) external skeletal fixation with autologous bone grafting. In the first stage, a thorough debridement and sequestrectomy were done. Antibiotic beads were used to obliterate the bone defect, and the wound was then directly closed. In the second stage, the bead chains were replaced with autogenous cancellous bone graft. Unilateral Hoffman external skeletal fixators were applied simultaneously. The mean follow-up period was 73.6 months (range, 29 months to 9 years). The length of time to achieve bony union ranged from 3.5 to 8 months (average, 4.3 months). Hoffman pin complication was found in two cases, which were then shifted to plate internal fixation. All the infections were eradicated, and the wounds healed without further skin graft or flap coverage. All the fractures achieved bony union except for one in a patient who died. Most patients acquired satisfactory function of elbow motion after removal of external fixation and physical therapy. The method of two-stage management was effective for infected humeral nonunion. Not only was the infection eradicated and osseous union achieved, but also the limb function and joint motion were preserved. PMID- 9390492 TI - Effect of heating on extracellular bioactive substances in stored human blood: in vitro study. AB - BACKGROUND: We have previously shown extracellular accumulation of various leukocyte and platelet-derived bioactive substances in human blood during storage. Release of bioactive substances may be temperature-dependent, and we studied the effect of heating during in vitro transfusion on bioactive substance accumulation in stored human blood. METHODS: Eight units of whole blood and eight units of prestorage leukofiltered whole blood were stored at 4 degrees C for 7 days. Subsequently, the blood from all 16 units was transfused via a blood heating device, which increased the blood temperature to 37 degrees C at outlet. Samples for enzyme-linked immunosorbent assay or radioimmunoassay analyses of histamine, myeloperoxidase (MPO), eosinophil cationic protein (ECP), and plasminogen activator inhibitor-1 (PAI-1) were drawn from the units at donation, after 7 days of storage just before transfusion, and during the in vitro transfusion. RESULTS: Extracellular concentrations of histamine, MPO, ECP, and PAI-1 were significantly (p < 0.05) increased in nonfiltered whole blood stored for 7 days compared with concentrations in fresh donated blood and in prestorage leukofiltered whole blood stored for 7 days. Heating reduced histamine, MPO, and ECP concentrations significantly (p < 0.05) in nonfiltered whole blood, whereas PAI-1 concentrations increased significantly (p < 0.05). Finally, there was no difference in concentrations of histamine, MPO, ECP, and PAI-1 in samples collected before and after heating of leukofiltered whole blood. CONCLUSIONS: Heating reduces accumulation of extracellular leukocyte-derived bioactive substances in whole blood, whereas it increases platelet-derived substances. Prestorage leukofiltration, however, reduces the obligatory extracellular accumulation of leukocyte and platelet-derived bioactive substances, which in addition is unchanged by heating. PMID- 9390493 TI - Optimized mesh expansion of composite skin grafts in rats treated with direct current. AB - BACKGROUND: The purpose of this study was to determine the optimum autoepidermal and allodermal expansion ratio of each component of a meshed composite skin graft (MCSG) that would lead to successful healing. METHODS: Male Sprague-Dawley rats were used as hosts of the MCSG and donors of autologous tissue. Male ACI rats were used as donors of allodermis. MCSGs with open meshed area (autoepidermal/allodermal) of 9:1/1.5:1, 9:1/3:1, 9:1/6:1, or 6:1/6:1 were applied to full-thickness skin defects and treated with a silver nylon dressing (SN) or SN with direct current (DC). Wound size, hair regrowth, and thickness of dermal layer were evaluated at 3 months. RESULTS: MCSGs of 9:1/1.5:1, 9:1/3:1, and 6:1/6:1 mesh ratios healed completely within 3 months with no difference in wound size between SN dressing groups or DC-treated groups. Application of DC reduced MCSG contraction and stimulated regrowth of hair. CONCLUSION: Fresh autoepidermis can be expanded 6:1 on a 6:1 allodermis or 9:1 on a 3:1 allodermis and achieve successful wound healing. PMID- 9390494 TI - Biodistribution of indocyanine green in a porcine burn model: light and fluorescence microscopy. AB - BACKGROUND: Infrared-excited fluorescence of intravenously administered indocyanine green (ICG) is being used as a method of early determination of burn depth. METHODS: Fluorescence microscopy and tissue fluorescence were recorded in a porcine burn model and correlated to burn severity and age. RESULTS: Recently placed superficial burns show significant fluorescence compared with adjacent normal tissue as a result of a strong inflammatory reaction in the superficial dermis with minimal vascular occlusion. The magnitude of the inflammatory reaction decreases with time. For deeper burns, vascular occlusion prevents transport of ICG into the burn and the intensity of ICG fluorescence in burn eschar is negligible. CONCLUSION: The intensity of ICG fluorescence measured at the surface of the wound for burns of similar age was shown to decrease exponentially with the depth of the burn. The enhanced fluorescence of partial thickness burns is attributable to increased permeability, and the decreased signal associated with deeper injuries is attributable to vascular occlusion. These results suggest that it is possible to differentiate burns that will heal spontaneously with minimal granulation from those that will not by measuring the intensity of ICG fluorescence. PMID- 9390495 TI - Practice patterns of pediatric surgeons caring for stable patients with traumatic solid organ injury. AB - BACKGROUND: Managed care financing has resulted in pressure to decrease hospital days and lower per diem costs. This influence may ultimately affect nonoperative management of blunt solid organ injuries in children (spleen, liver, kidneys). METHODS: Pediatric surgeons caring for trauma patients were surveyed regarding current practice patterns. One survey was sent to a representative staff pediatric surgeon at each major children's hospital or children's unit involved in the care of the injured child in the United States. RESULTS: There were 87 responses to 117 surveys (75%). Relatively few children fail nonoperative management. For major management decisions, including radiographic study of choice; when to transfuse; and when to allow out of bed, home, and back to school, there was often a clear majority opinion of appropriate care. However, there was a wide variance in response for some questions. CONCLUSIONS: Surgical judgment must be individualized, but a low number of failures of nonoperative management is helpful in delineating safe practice guidelines. Surgeons using fewer resources than the norm may help delineate management schemes that are equally effective to more expensive care. Based on these responses a management protocol is recommended. PMID- 9390496 TI - Geographic variation in serious nonfatal firearm injuries in Pennsylvania. AB - OBJECTIVE: The purpose of this study was to characterize the geographic epidemiology of serious nonfatal firearm injuries (NFFI) within Pennsylvania during a 6-year period. METHODS: A historical review of data from the Pennsylvania Trauma System Foundation trauma registry was completed using county level data. Based on a format adapted from the United States Department of Agriculture, NFFI in Pennsylvania were classified by their county of occurrence: central city counties, metropolitan counties, nonmetropolitan counties, or rural counties. Population-based rates of NFFI were then calculated, as were NFFI as a proportion of the number of injuries within each region. These rates were stratified by intent of injury, scene of injury, and type of firearm. RESULTS: A total of 100,703 trauma cases were reported to the Pennsylvania Trauma System Foundation from 1988 through 1993, of which 5,847 were serious NFFI. Nonfatal firearm assaults increased from rural counties to central city counties, whereas unintentional NFFI decreased (p < 0.05). A 225% increase in the number of NFFI, from 445 cases in 1988 to 1,004 cases in 1993, was noted in the central city counties. Comparatively, the increase in the noncity regions was 145%, from 182 cases in 1988 to 263 in 1993. Nonfatal firearm injuries occurred most often at home in noncity counties (rural, nonmetropolitan, and metropolitan counties) (47.9%). This is in contrast to central city counties, where NFFI occurred significantly more often in the street (53.5%) (p < 0.05). Handgun NFFI increased, whereas rifle NFFI decreased, from rural counties to central city counties (p < 0.05). Relative to population size, the risk of shotgun injuries was greatest in central city counties and lowest in rural counties. Shotgun injuries also accounted for a significantly longer hospital stay (15.06 days) compared with handgun injuries (10.38 days) and rifle injuries (11.81 days) (p < 0.05). CONCLUSION: Significant variation in NFFI was observed across population based regions in Pennsylvania. Rural areas demonstrated relatively high risks of NFFI committed unintentionally, in the home, and with rifles. As regional populations increase, relatively high risks of NFFI, committed as assaults, in the street, and by handguns, are highlighted. Although handguns were the most prominent firearm associated with NFFI, nonfatal shotgun injuries produced substantially longer hospital stays and may be an underappreciated cause of nonfatal firearm assaults in the urban setting. PMID- 9390497 TI - Reproducibility of preventable death judgments and problem identification in 60 consecutive road trauma fatalities in Victoria, Australia. Consultative Committee on Road Traffic Fatalities in Victoria. AB - BACKGROUND: Since 1992, the Consultative Committee on Road Traffic Fatalities in Victoria has identified problems in the management of traffic fatalities. Its two evaluative committees have additionally assessed the potential preventability of death. Previous studies have shown only poor to fair reproducibility of death judgments. METHODS: Problems in the management of 60 consecutive road traffic fatalities and the potential preventability of death were independently evaluated by the two committees. Inter-rater and inter-committee concordance were analyzed using the kappa statistic. RESULTS: Reproducibility was high. Inter-committee agreement on nonpreventable, potentially preventable, and preventable death judgments was high (kappa = 0.73, 95% confidence interval = 0.57-0.89). Agreement within the two evaluative committees was also high (average weighted kappa = 0.73 and 0.74). There was good agreement between committees on problems identified, including those contributing to death. CONCLUSION: The high kappa concordance on preventable death judgments and the agreement on problem identification supports the reproducibility of the methodology used. PMID- 9390498 TI - Bilateral craniotomies for blunt head trauma. AB - Development of delayed or recurrent intracranial hematomas requiring reexploration or a secondary craniotomy is well known. Patients with bilateral pathology requiring bilateral craniotomies as the initial emergency operative intervention, however, are uncommon. The lack of available literature and the large volume of head trauma seen at our institution prompted us to analyze the retrospective data on blunt head injury requiring bilateral craniotomies. Twenty patients underwent bilateral craniotomies at the University of Miami/Jackson Memorial Medical Center between January 1986 and June 1994. Ages ranged from 18 to 85 years. Mechanism of injury included motor vehicle crash (n = 4), pedestrian hit by automobile (n = 4), assault (n = 8), fall from height (n = 3), and unknown (n = 1). Epidural hematomas, acute subdural hematomas, contusions, and intracerebral hematomas were seen in varying combinations. The preoperative Glasgow Coma Scale (GCS) score ranged from 4 to 14, with a mean of 8.8 (+/-0.82 SE). Sixteen of the 20 patients survived and were discharged from the hospital. The survivors' Rancho Los Amigos Scale score on discharge ranged from 2 to 8, with a mean of 6.1 (+/-0.45 SE). A Fisher's exact test was performed to compare the outcome between the patients with mild (GCS score 13-15) to moderate (GCS score 9-12) head injury and those with severe (GCS score 4-8) head injury. It showed a statistically higher frequency of death in the severe category (p < 0.05). In conclusion, the outcome of patients with bilateral pathology requiring emergency bilateral craniotomy at initial treatment correlated well with their GCS scores at initial presentation. PMID- 9390499 TI - Outcome of blunt thoracic aortic injury in a level I trauma center: an 8-year review. AB - BACKGROUND: The purpose of this study was to evaluate our experience with blunt thoracic aortic injury and identify factors predictive of outcome. METHODS: Hospital charts, trauma registry data, and autopsies of 64 patients with blunt thoracic aortic injury from 1988 to 1995 were reviewed. RESULTS: Patients were identified and segregated based on admission physiology. Group 1 patients (n = 19) arrived in arrest. Group 2 patients (n = 10) arrived in shock with systolic BP 90. Group 3 patients (n = 35) arrived with systolic BP>90. All patients in groups 1 and 2 expired. Injury Severity Scores for nonsurvivors in group 3 (n = 12) were significantly higher than survivors. There were no significant differences when comparing time of injury to repair or arrival between groups, or in mortality or paralysis comparing repair techniques or clamp/bypass times. Double lumen endotracheal tubes caused significant operative delays compared to single lumen tubes. CONCLUSIONS: Predictors of survivability were hemodynamic stability on arrival and lower Injury Severity Scores. In thoracic aortic injury patients arriving hemodynamically stable, Injury Severity Score correlated with mortality but not paralysis. PMID- 9390500 TI - Splanchnic ischemia and bacterial translocation in the abdominal compartment syndrome. AB - BACKGROUND AND METHODS: Major trauma or abdominal injury may lead to the development of increased intra-abdominal pressure (IAP) and the onset of the abdominal compartment syndrome. Although the effect of raised IAP on systemic and splanchnic hemodynamics have been described, the consequences of the resultant gut hypoperfusion in this setting are unknown. Bacterial translocation (BT) occurs after a period of splanchnic ischemia and may contribute to later organ failure. A rodent model was used to examine the effect of raised IAP on ileal mucosal blood flow (MBF) and BT. IAP was increased to 25 mm Hg for 60 minutes and mean arterial blood pressure was maintained with fluid. Animals were killed 24 hours later and examined for BT. RESULTS: Increased IAP resulted in a decrease of MBF to 63% of baseline despite maintaining normal mean arterial blood pressure. BT occurred principally to the mesenteric lymph nodes after 60 minutes of IAP at 25 mm Hg. CONCLUSIONS: Increased IAP leads to decreased MBF and to BT, which may contribute to later septic complications and organ failure. PMID- 9390501 TI - A case of deep laceration of the lung treated with video-assisted thoracic surgical lobectomy: case report. PMID- 9390502 TI - Three-year follow-up of a posttraumatic right coronary aneurysm. AB - Posttraumatic saccular aneurysm of the right coronary artery is a rare complication of nonpenetrating chest trauma. We observed a posttraumatic coronary aneurysm for 3 years and noted that the aneurysm has changed in shape, with partial obliteration of the aneurysm sac, and that its clinical course was uneventful with conservative treatment. Surgical removal of aneurysms has been advocated in the literature; however, conservative medical treatment and a wait and-see policy can be considered as a treatment modality for posttraumatic coronary aneurysm. PMID- 9390503 TI - Aortoiliac dissection after blunt abdominal trauma: case report. AB - The distal abdominal aorta is rarely injured after blunt trauma but a direct blow to the abdomen from a seatbelt or handlebars may cause intimal dissection or rupture. We present the diagnosis and surgical management of aortoiliac dissection in a 16-year-old boy injured in a motorcycle accident. The technical aspects of vascular repair are emphasized. PMID- 9390504 TI - Intrathoracic migration of a Kirschner wire: case report. PMID- 9390505 TI - Pelvic injuries in equestrians on buck-jumping horses. PMID- 9390506 TI - Laterally based skin flap for below-knee amputation: case report. PMID- 9390507 TI - How hearing happens. PMID- 9390508 TI - Fish n' chicks: model recipes for hair-cell regeneration? PMID- 9390509 TI - Myosin and adaptation by hair cells. PMID- 9390510 TI - Encoding of timing in the brain stem auditory nuclei of vertebrates. PMID- 9390511 TI - Distributed time-domain representations in the birdsong system. PMID- 9390512 TI - A turning point in schizophrenia genetics. PMID- 9390513 TI - Xath5 participates in a network of bHLH genes in the developing Xenopus retina. AB - We examined the function of basic-helix-loop-helix (bHLH) transcription factors during retinal neurogenesis. We identified Xath5, a Xenopus bHLH gene related to Drosophila atonal, which is expressed in the developing Xenopus retina. Targeted expression of Xath5 in retinal progenitor cells biased the differentiation of these cells toward a ganglion cell fate, suggesting that Xath5 can regulate the differentiation of retinal neurons. We examined the relationship between the three bHLH genes Xash3, NeuroD, and Xath5 during retinal neurogenesis and found that Xash3 is expressed in early retinoblasts, followed by coexpression of Xath5 and NeuroD in differentiating cells. We provide evidence that the expression of Xash3, NeuroD, and Xath5 is coupled and propose that these bHLH genes regulate successive stages of neuronal differentiation in the developing retina. PMID- 9390514 TI - Neuropilin-semaphorin III/D-mediated chemorepulsive signals play a crucial role in peripheral nerve projection in mice. AB - Neuropilin is a neuronal cell surface protein and has been shown to function as a receptor for a secreted protein, semaphorin III/D, that can induce neuronal growth cone collapse and repulsion of neurites in vitro. The roles of neuropilin in vivo, however, are unknown. Here, we report that neuropilin-deficient mutant mice produced by targeted disruption of the neuropilin gene show severe abnormalities in the trajectory of efferent fibers of the PNS. We also describe that neuropilin-deprived dorsal root ganglion neurons are perfectly protected from growth cone collapse elicited by semaphorin III/D. Our results indicate that neuropilin-semaphorin III/D-mediated chemorepulsive signals play a major role in guidance of PNS efferents. PMID- 9390515 TI - Synaptic clustering of Fascilin II and Shaker: essential targeting sequences and role of Dlg. AB - Previous studies have shown that both the Fasciclin II (Fas II) cell adhesion molecule and the Shaker potassium channel are localized at the Drosophila neuromuscular junction, where they function in the growth and plasticity of the synapse. Here, we use the GAL4-UAS system to drive expression of the chimeric proteins CD8-Fas II and CD8-Shaker and show that the C-terminal sequences of both Fas II and Shaker are necessary and sufficient to drive the synaptic localization of a heterologous protein. Moreover, we show that the PDZ-containing protein Discs-Large (Dlg) controls the localization of these proteins, most likely through a direct interaction with their C-terminal amino acids. Finally, transient expression studies show that the pathway these proteins take to the synapse involves either an active clustering or a selective stabilization in the synaptic membrane. PMID- 9390516 TI - Crx, a novel Otx-like paired-homeodomain protein, binds to and transactivates photoreceptor cell-specific genes. AB - The otd/Otx gene family encodes paired-like homeodomain proteins that are involved in the regulation of anterior head structure and sensory organ development. Using the yeast one-hybrid screen with a bait containing the Ret 4 site from the bovine rhodopsin promoter, we have cloned a new member of the family, Crx (Cone rod homeobox). Crx encodes a 299 amino acid residue protein with a paired-like homeodomain near its N terminus. In the adult, it is expressed predominantly in photoreceptors and pinealocytes. In the developing mouse retina, it is expressed by embryonic day 12.5 (E12.5). Recombinant Crx binds in vitro not only to the Ret 4 site but also to the Ret 1 and BAT-1 sites. In transient transfection studies, Crx transactivates rhodopsin promoter-reporter constructs. Its activity is synergistic with that of Nrl. Crx also binds to and transactivates the genes for several other photoreceptor cell-specific proteins (interphotoreceptor retinoid-binding protein, beta-phosphodiesterase, and arrestin). Human Crx maps to chromosome 19q13.3, the site of a cone rod dystrophy (CORDII). These studies implicate Crx as a potentially important regulator of photoreceptor cell development and gene expression and also identify it as a candidate gene for CORDII and other retinal diseases. PMID- 9390517 TI - CREB: a major mediator of neuronal neurotrophin responses. AB - Neurotrophins regulate neuronal survival, differentiation, and synaptic function. To understand how neurotrophins elicit such diverse responses, we elucidated signaling pathways by which brain-derived neurotrophic factor (BDNF) activates gene expression in cultured neurons and hippocampal slices. We found, unexpectedly, that the transcription factor cyclic AMP response element-binding protein (CREB) is an important regulator of BDNF-induced gene expression. Exposure of neurons to BDNF stimulates CREB phosphorylation and activation via at least two signaling pathways: by a calcium/calmodulin-dependent kinase IV (CaMKIV)-regulated pathway that is activated by the release of intracellular calcium and by a Ras-dependent pathway. These findings reveal a previously unrecognized, CaMK-dependent mechanism by which neurotrophins activate CREB and suggest that CREB plays a central role in mediating neurotrophin responses in neurons. PMID- 9390518 TI - Localized changes in immediate-early gene regulation during sensory and motor learning in zebra finches. AB - A complex neural system controls birdsong learning, but its organization is not understood, nor is it known why learning only occurs during a critical period in adolescence. Here, we analyzed developmental regulation in zebra finches of zenk, an immediate-early gene (IEG) implicated in memory consolidation. Basal expression was elevated within auditory telencephalon (specifically, within the caudomedial neostriatum [NCM]) during song acquisition. Expression could be further induced by song playbacks 30 days after hatching but not at 20 days nor in juveniles reared in severe isolation. Singing itself induced zenk in song production nuclei, including Area X, even in adults. Within a compartment of the robust nucleus of the archistriatum (RA), however, this response dwindled as singing matured. These results suggest that the onset of sensory memory storage may be regulated in part at NCM, and motor plasticity may be regulated at RA. PMID- 9390519 TI - Distribution of Ca2+-activated K+ channel isoforms along the tonotopic gradient of the chicken's cochlea. AB - In some cochleae, the number and kinetic properties of Ca2+-activated K+ (KCa) channels partly determine the characteristic frequency of each hair cell and thus help establish a tonotopic map. In the chicken's basilar papilla, we found numerous isoforms of KCa channels generated by alternative mRNA splicing at seven sites in a single gene, cSlo. In situ polymerase chain reactions demonstrated cSlo expression in hair cells and revealed differential distributions of KCa channel isoforms along the basilar papilla. Analysis of single hair cells by the reverse transcription polymerase chain reaction confirmed the differential expression of channel variants. Heterologously expressed cSlo variants differed in their sensitivities to Ca2+ and voltage, suggesting that the distinct spatial distributions of cSlo variants help determine the tonotopic map. PMID- 9390520 TI - Differential distribution of Ca2+-activated K+ channel splice variants among hair cells along the tonotopic axis of the chick cochlea. AB - We have cloned from the receptor epithelium of the chick cochlea a family of alternatively spliced cDNAs derived from cslo, which encodes a Ca2+-activated K+ channel like those shown to help determine the resonant frequency of electrically tuned hair cells. Our results from PCRs using template RNAs from both tonotopically subdivided receptor epithelia and single hair cells demonstrate differential exon usage along the frequency axis of the epithelium at multiple splice sites in cslo. We also show that single hair cells express more than one splice variant at a given splice site. Since channel isoforms encoded by differentially spliced slo transcripts in other species are functionally heterogeneous, these data suggest that differential processing of slo transcripts may account, at least in part, for the systematic variation in hair-cell membrane properties along the frequency axis of electrically tuned auditory receptor epithelia. PMID- 9390521 TI - Structural organization of the synaptic exocytosis core complex. AB - Syntaxin, vesicle-associated membrane protein (VAMP), and synaptosome-associated protein of 25 kDa (SNAP-25) form a ternary "core complex" central to the process of synaptic vesicle docking and fusion. Several lines of evidence support the hypothesis that the proteins assemble in a coiled-coil structure, but the alignment of alpha helices in this coil and the overall conformation of the coil are unknown. We employ the technique of fluorescence resonance energy transfer (FRET) to investigate the alignment between syntaxin and VAMP. With the acceptor probe coupled to the amino-terminal end of the VAMP coiled-coil domain, the donor probe fluorescence is quenched to a greater extent when it is on the amino terminal end of the syntaxin H3 domain than when it is on the carboxy-terminal end. The data indicate that syntaxin and VAMP bind primarily in a parallel arrangement and suggest a coiled-coil structure that is bent rather than fully extended. We propose a model in which binding of SNAP receptor (SNARE) protein coiled-coil domains helps drive vesicle fusion. PMID- 9390522 TI - Neuronal peptide release is limited by secretory granule mobility. AB - Neuropeptides are slowly released from a limited pool of secretory granules. To visualize this process, GFP-tagged preproatrial natriuretic factor (ANF) was expressed in nerve growth factor-treated PC12 cells. Biochemical and microfluorimetric experiments demonstrate that proANF-EGFP is packaged in granules that accumulate at neurite endings and is released in a Ca2+-dependent manner by secretagogs. Confocal microscopy shows that secretion is associated with depletion of granules distributed throughout the terminal. Fluorescence recovery after photobleaching and time-lapse particle tracking reveal that only a subpopulation of cytoplasmic secretory granules, similar in size to the releasable pool, can move quickly enough (D = 6 x 10(-11) cm2/s) to support release. Therefore, sustained secretory responses are limited by the number of mobile granules and their slow rate of diffusion. PMID- 9390523 TI - Plasticity in fast synaptic inhibition of adult oxytocin neurons caused by switch in GABA(A) receptor subunit expression. AB - We found that magnocellular oxytocin neurons in adult female rats exhibit an endogenous GABA(A) receptor subunit switch around parturition: a decrease in alpha1:alpha2 subunit mRNA ratio correlated with a decrease in allopregnanolone potentiation and increase in decay time constant of the GABA(A) receptor-mediated IPSCs in these cells. The causal relationship between changes in alpha1:alpha2 mRNA ratio and the ion channel kinetics was confirmed using in vitro antisense deletion. Further, GABA(A) receptors exhibited a tonic inhibitory influence upon oxytocin release in vivo, and allopregnanolone helped to restrain oxytocin neuron in vitro firing only before parturition, when the alpha1:alpha2 subunit mRNA ratio was still high. Such observations provide evidence for the physiological significance of GABA(A) receptor subunit heterogeneity and plasticity in the adult brain. PMID- 9390524 TI - D5 dopamine receptors enhance Zn2+-sensitive GABA(A) currents in striatal cholinergic interneurons through a PKA/PP1 cascade. AB - Cholinergic interneurons have been implicated in striatally mediated associative learning. In classical conditioning paradigms, conditioned stimuli trigger a transient suppression of neuronal activity that is dependent upon an intact dopaminergic innervation. Our hypothesis was that this suppression reflected dopaminergic enhancement of sensory-linked GABAergic input. As a test, the impact of dopamine on interneuronal GABA(A) receptor function was studied by combined patch-clamp recording and single-cell reverse transcription PCR. Activation of D5 dopamine receptors reversibly enhanced a Zn2+-sensitive component of GABA(A) currents. Although dependent upon protein kinase A (PKA) activation, the modulation was blocked by protein phosphatase 1 (PP1) inhibition, suggesting it was dependent upon dephosphorylation. These results establish a novel mechanism by which intrastriatally released dopamine mediates changes in GABAergic signaling that could underlie the initial stages of associative learning. PMID- 9390525 TI - Characterizing voltage-dependent conformational changes in the Shaker K+ channel with fluorescence. AB - We examined voltage-dependent conformational changes in three specific regions of the Shaker potassium channel with site-directed fluorescent labeling: the fourth transmembrane segment (S4), the second transmembrane segment (S2), and the putative pore region. The fluorescence changes displayed distinctive properties that correlate with gating, activation, and slow inactivation of the channel. The fluorescence signals measured near the S2 and S4 segments suggest that the S2 segment may undergo voltage-sensitive conformational changes that precede those in the S4 segment. In contrast, fluorescence changes in the pore correlated with the voltage dependence and time course of ionic activation and slow inactivation. Spectroscopy indicated that the mechanism of fluorescence change involves voltage dependent quenching of the probe in an aqueous environment by other parts of the protein. PMID- 9390526 TI - Rat GluR7 and a carboxy-terminal splice variant, GluR7b, are functional kainate receptor subunits with a low sensitivity to glutamate. AB - Glutamate receptors of the kainate-preferring subtype have recently been shown to mediate synaptic transmission in the hippocampus. The low-affinity kainate receptor subunit GluR7 was found to be nonfunctional in previous studies. We report here that the GluR7 subunit and a novel carboxy-terminal splice variant, GluR7b, are functional glutamate receptors with unique pharmacological properties. In particular, glutamate exhibits a 10-fold lower potency for (non desensitized) GluR7-mediated currents as compared to other non-NMDA receptor channels. These data will facilitate understanding of the distinct role played by GluR7 receptors in synaptic transmission. PMID- 9390528 TI - Heterogeneity of health maintenance organizations and quality of care. PMID- 9390527 TI - Adjuvant therapy for early breast cancer: a time to refine. PMID- 9390529 TI - Association of oral cancers with alcohol consumption: exploring mechanisms. PMID- 9390530 TI - Angiogenesis: the unifying concept in cancer? PMID- 9390531 TI - Apoptosis research is yielding many potential drug targets. PMID- 9390532 TI - CNS consortia reshaping brain tumor research. PMID- 9390533 TI - Exposing the defenses of gliomas. PMID- 9390534 TI - HPV vaccines for cervical cancer move toward clinic, encounter social issues. PMID- 9390535 TI - Meeting report: genetic environmental interactions in cancer susceptibility in animal models. PMID- 9390536 TI - Tamoxifen and chemotherapy for lymph node-negative, estrogen receptor-positive breast cancer. AB - BACKGROUND: The B-20 study of the National Surgical Adjuvant Breast and Bowel Project (NSABP) was conducted to determine whether chemotherapy plus tamoxifen would be of greater benefit than tamoxifen alone in the treatment of patients with axillary lymph node-negative, estrogen receptor-positive breast cancer. METHODS: Eligible patients (n = 2306) were randomly assigned to one of three treatment groups following surgery. A total of 771 patients with follow-up data received tamoxifen alone; 767 received methotrexate, fluorouracil, and tamoxifen (MFT); and 768 received cyclophosphamide, methotrexate, fluorouracil, and tamoxifen (CMFT). The Kaplan-Meier method was used to estimate disease-free survival, distant disease-free survival, and survival. Reported P values are two sided. RESULTS: Through 5 years of follow-up, chemotherapy plus tamoxifen resulted in significantly better disease-free survival than tamoxifen alone (90% for MFT versus 85% for tamoxifen [P = .01]; 89% for CMFT versus 85% for tamoxifen [P = .001]). A similar benefit was observed in both distant disease-free survival (92% for MFT versus 87% for tamoxifen [P = .008]; 91% for CMFT versus 87% for tamoxifen [P = .006]) and survival (97% for MFT versus 94% for tamoxifen [P = .05]; 96% for CMFT versus 94% for tamoxifen [P = .03]). Compared with tamoxifen alone, MFT and CMFT reduced the risk of ipsilateral breast tumor recurrence after lumpectomy and the risk of recurrence at other local, regional, and distant sites. Risk of treatment failure was reduced after both types of chemotherapy, regardless of tumor size, tumor estrogen or progesterone receptor level, or patient age; however, the reduction was greatest in patients aged 49 years or less. No subgroup of patients evaluated in this study failed to benefit from chemotherapy. CONCLUSIONS: Findings from this and other NSABP studies indicate that patients with breast cancer who meet NSABP protocol criteria, regardless of age, lymph node status, tumor size, or estrogen receptor status, are candidates for chemotherapy. PMID- 9390537 TI - Breast cancer survival and treatment in health maintenance organization and fee for-service settings. AB - BACKGROUND: Enrollment in health maintenance organizations (HMOs) has increased rapidly during the past 10 years, reflecting a growing emphasis on health care cost containment. To determine whether there is a difference in the treatment and outcome for female patients with breast cancer enrolled in HMOs versus a fee-for service setting, we compared the 10-year survival and initial treatment of patients with breast cancer enrolled in both types of plans. METHODS: With the use of tumor registries covering the greater San Francisco-Oakland and Seattle Puget Sound areas, respectively, we obtained information on the treatment and outcome for 13,358 female patients with breast cancer, aged 65 years and older, diagnosed between 1985 and 1992. We linked registry information with Medicare data and data from the two large HMOs included in the study. We compared the survival and treatment differences between HMO and fee-for-service care after adjusting for tumor stage, comorbidity, and sociodemographic characteristics. RESULTS: In San Francisco-Oakland, the 10-year adjusted risk ratio for breast cancer deaths among HMO patients compared with fee-for-service patients was 0.71 (95% confidence interval [CI] = 0.59-0.87) and was comparable for all deaths. In Seattle-Puget Sound, the risk ratio for breast cancer deaths was 1.01 (95% CI = 0.77-1.33) but somewhat lower for all deaths. Women enrolled in HMOs were more likely to receive breast-conserving surgery than women in fee-for-service (odds ratio = 1.55 in San Francisco-Oakland; 3.39 in Seattle). HMO enrollees undergoing breast-conserving surgery were also more likely to receive adjuvant radiotherapy (San Francisco-Oakland odds ratio = 2.49; Seattle odds ratio = 4.62). CONCLUSIONS: Long-term survival outcomes in the two prepaid group practice HMOs in this study were at least equal to, and possibly better than, outcomes in the fee-for-service system. In addition, the use of recommended therapy for early stage breast cancer was more frequent in the two HMOs. PMID- 9390538 TI - Effects of acetaldehyde on cell regeneration and differentiation of the upper gastrointestinal tract mucosa. AB - BACKGROUND: The tumor-promoting effect of ethanol on cancer of the upper respiratory-digestive tract is not well understood. Although ethanol itself is not carcinogenic, the first product of ethanol metabolism-acetaldehyde is. Acetaldehyde can be produced from ethanol by oral bacteria, and high concentrations have been observed in human saliva after ethanol consumption. The purpose of this study was to investigate whether acetaldehyde administered orally to rats induces altered differentiation and proliferation in the animals' upper gastrointestinal tracts. METHODS: Twenty Wistar rats were given either water containing acetaldehyde at a concentration of 120 mM or tap water to drink for 8 months. Tissue specimens were then taken from the tongue, epiglottis, and forestomach of each animal and immunohistochemically stained for markers of cellular proliferation (Ki67 nuclear antigen) or differentiation (cytokeratins 1, 4, 10, 11, 14, and 19). The mean epithelial thickness of each sample was measured via light microscopy, using an eyepiece containing grid lines. Differences between the control and acetaldehyde-treated groups were analyzed by use of the unpaired Student's t test. All reported P values are two-sided. RESULTS: Although no tumors were observed, staining for cytokeratins 4 and 14 revealed an enlarged basal layer of squamous epithelia in the rats receiving acetaldehyde. In these animals, cell proliferation was significantly greater than that observed in the control animals for samples from the tongue (P<.0001), epiglottis (P<.001), and forestomach (P<.0001). In addition, the epithelia from acetaldehyde-treated rats were significantly thicker than in epithelia from control animals (P<.05 for all three sites). CONCLUSIONS: Acetaldehyde, administered orally to rats, can cause hyperplastic and hyperproliferative changes in epithelia of the upper gastrointestinal tract. This finding suggests that microbially produced acetaldehyde in saliva may explain the tumor-promoting effect of ethanol on these epithelia. PMID- 9390540 TI - P-glycoprotein expression: critical determinant in the response to osteosarcoma chemotherapy. AB - BACKGROUND: Fewer than 20% of patients with bone cancer who are treated with surgery alone are cured. Even with the best current treatment, surgery combined with chemotherapy, only 60%-80% of patients with nonmetastatic bone cancer and 10% of patients with metastatic bone cancer are cured. Thus far, the reason for treatment failure in the nonresponding subset has not been identified. It has been hypothesized that P-glycoprotein, which confers multidrug resistance, might be the cause. We sought to determine whether the expression of P-glycoprotein is associated with poor treatment outcome in osteosarcoma. METHODS: In a retrospective study, we correlated P-glycoprotein expression with the outcome of conventional chemotherapy in 62 consecutive, clinically staged patients diagnosed as having osteosarcoma between 1980 and 1989. RESULTS: P-glycoprotein was overexpressed in 27 patients but not in another 34 patients, and expression was ambiguous in the sample from one patient. At a median follow-up of 8.9 years, the 34 patients whose tumors did not express P-glycoprotein had significantly better relapse-free rates than the 27 subjects whose tumors expressed the protein (87% versus 0%; P<.00001) and had improved survival rates (94% versus 35%; P<.00001). Among the 46 patients who received chemotherapy before surgery, the 23 whose tumors were negative for P-glycoprotein showed significantly better long-term outcomes (P<.00002), although differences in tumor necrosis in response to therapy were only of borderline significance (P = .057). CONCLUSIONS: P glycoprotein expression does correlate with treatment failure in patients with osteosarcoma. This correlation raises the possibility that inhibiting the action of P-glycoprotein as part of therapy for this disease would improve outcome. PMID- 9390539 TI - Alcohol dehydrogenase 3 genotype and risk of oral cavity and pharyngeal cancers. AB - BACKGROUND: The consumption of alcoholic beverages is a strong risk factor for cancers of the oral cavity and pharynx (oral cancers). Alcohol dehydrogenase type 3 (ADH3) metabolizes ethanol to acetaldehyde, a carcinogen. We evaluated whether individuals homozygous for the fast-metabolizing ADH3(1) allele (ADH3[1-1]) have a greater risk of developing oral cancer in the presence of alcoholic beverage consumption than those with the slow-metabolizing ADH3(2) allele (ADH3[1-2] and ADH3[2-2]). METHODS: As part of a population-based study of oral cancer conducted in Puerto Rico, the ADH3 genotypes of 137 patients with histologically confirmed oral cancer and of 146 control subjects (i.e., individuals with no history of oral cancer) were determined by molecular genetic analysis of oral epithelial cell samples. Risks were estimated by use of multiple logistic regression analyses. RESULTS: Compared with nondrinkers with the ADH3(1-1) genotype, consumers of at least 57 alcoholic drinks per week with the ADH3(1-1), ADH3(1-2), and ADH3(2-2) genotypes had 40.1-fold (95% confidence interval [CI] = 5.4-296.0), 7.0-fold (95% CI = 1.4-35.0), and 4.4-fold (95% CI = 0.6-33.0) increased risks of oral cancer, respectively; the risk associated with the ADH3(1-1) genotype, compared with the ADH3(1-2) and ADH3(2-2) genotypes combined, was 5.3 (95% CI = 1.0-28.8) among such drinkers. Considering all levels of alcohol consumption, the risk of oral cancer per additional alcoholic drink per week increased 3.6% (95% CI = 1.9%-5.4%) for subjects with the ADH3(1-1) genotype and 2.0% (95% CI = 0.9% 3.0%) for subjects with the ADH3(1-2) or ADH3(2-2) genotype (two-sided P = .04). CONCLUSIONS: The ADH3(1-1) genotype appears to substantially increase the risk of ethanol-related oral cancer, thus providing further evidence for the carcinogenicity of acetaldehyde. PMID- 9390542 TI - Breast density and cancer. PMID- 9390541 TI - Serum levels of prostate-specific antigen among Japanese-American and native Japanese men. AB - BACKGROUND: Fourfold to sixfold higher prostate cancer rates in Japanese-American men in the United States compared with Japanese men in Japan have been cited to support a role for environmental risk factors in the etiology of the disease. To examine the hypothesis that part or all of the elevated prostate cancer rates in Japanese-American men may reflect more intensive prostate cancer screening in the United States than in Japan, we compared prostate-specific antigen (PSA) levels in community-based samples of serum from men without prostate cancer. METHODS: Japanese-American men aged 40-85 years and native Japanese men aged 40-89 years with no history of prostate cancer provided sera, respectively, in the United States from March 1990 through March 1992 (n = 237) or in Japan from January 1992 through December 1993 (n = 3522). Age-specific PSA levels were used to estimate the prevalences of undetected prostate cancer in the two populations. RESULTS: Age-specific mean PSA levels were significantly lower in Japanese-Americans than in native Japanese (two-sided P<.001). The prevalence of an elevated PSA level increased with age in both populations and exceeded 5% among men aged 60 years or more. Combined with data on prevalence of detected prostate cancer in the two populations, our data suggest that some 10.0% of Japanese-Americans aged 75 years have prostate cancer, with 31% of that fraction remaining undiagnosed. The corresponding estimates in Japan are a total cancer prevalence of 5.4%, of which 81% has not been detected clinically. CONCLUSIONS: The total cancer prevalence ratio 10.0/5.4 = 1.9 (95% confidence interval = 1.5-2.3) in Japanese-American men compared with Japanese men in Japan suggests an increased risk for Japanese American men, but of less magnitude than the fourfold to sixfold increase indicated by the incidence data. PMID- 9390543 TI - Re: Risk factors for lung cancer and for intervention effects in CARET, the Beta Carotene and Retinol Efficacy Trial. PMID- 9390544 TI - Re: saturated fat intake and lung cancer risk among nonsmoking women in Missouri. PMID- 9390545 TI - Re: Correlating nutrition to recent cancer mortality statistics. PMID- 9390546 TI - Re: Relationship between lifetime ovulatory cycles and overexpression of mutant p53 in epithelial ovarian cancer. PMID- 9390547 TI - Re: Carcinogenicity of the drinking water mutagen 3-chloro-4-(dichloromethyl)-5 hydroxy-2(5H)-furanone in the rat. PMID- 9390548 TI - Re: Benzene and the dose-related incidence of hematologic neoplasms in China. PMID- 9390549 TI - Viral proteases: evolution of diverse structural motifs to optimize function. PMID- 9390550 TI - Protein translocation channels in the proteasome and other proteases. PMID- 9390551 TI - Regulatory subunits of energy-dependent proteases. PMID- 9390552 TI - ECM and cell surface proteolysis: regulating cellular ecology. PMID- 9390553 TI - Caspases: intracellular signaling by proteolysis. PMID- 9390554 TI - The structure of ClpP at 2.3 A resolution suggests a model for ATP-dependent proteolysis. AB - We have determined the crystal structure of the proteolytic component of the caseinolytic Clp protease (ClpP) from E. coli at 2.3 A resolution using an ab initio phasing procedure that exploits the internal 14-fold symmetry of the oligomer. The structure of a ClpP monomer has a distinct fold that defines a fifth structural family of serine proteases but a conserved catalytic apparatus. The active protease resembles a hollow, solid-walled cylinder composed of two 7 fold symmetric rings stacked back-to-back. Its 14 proteolytic active sites are located within a central, roughly spherical chamber approximately 51 A in diameter. Access to the proteolytic chamber is controlled by two axial pores, each having a minimum diameter of approximately 10 A. From the structural features of ClpP, we suggest a model for its action in degrading proteins. PMID- 9390555 TI - The exosome: a conserved eukaryotic RNA processing complex containing multiple 3' ->5' exoribonucleases. AB - We identified a complex in S. cerevisiae, the "exosome," consisting of the five essential proteins Rrp4p, Rrp41p, Rrp42p, Rrp43p, and Rrp44p (Dis3p). Remarkably, four of these proteins are homologous to characterized bacterial 3'-->5' exoribonucleases; Rrp44p is homologous to RNase II, while Rrp41p, Rrp42p, and Rrp43p are related to RNase PH. Recombinant Rrp4p, Rrp44p, and Rrp41p are 3'-->5' exoribonucleases in vitro that have distributive, processive, and phosphorolytic activities, respectively. All components of the exosome are required for 3' processing of the 5.8S rRNA. Human Rrp4p is found in a comparably sized complex, and expression of the hRRP4 gene in yeast complements the rrp4-1 mutation. We conclude that the exosome constitutes a highly conserved eukaryotic RNA processing complex. PMID- 9390556 TI - Mutation in the mismatch repair gene Msh6 causes cancer susceptibility. AB - Mice carrying a null mutation in the mismatch repair gene Msh6 were generated by gene targeting. Cells that were homozygous for the mutation did not produce any detectable MSH6 protein, and extracts prepared from these cells were defective for repair of single nucleotide mismatches. Repair of 1, 2, and 4 nucleotide insertion/deletion mismatches was unaffected. Mice that were homozygous for the mutation had a reduced life span. The mice developed a spectrum of tumors, the most predominant of which were gastrointestinal tumors and B- as well as T-cell lymphomas. The tumors did not show any microsatellite instability. We conclude that MSH6 mutations, like those in some other members of the family of mismatch repair genes, lead to cancer susceptibility, and germline mutations in this gene may be associated with a cancer predisposition syndrome that does not show microsatellite instability. PMID- 9390557 TI - Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade. AB - We report here the purification of the third protein factor, Apaf-3, that participates in caspase-3 activation in vitro. Apaf-3 was identified as a member of the caspase family, caspase-9. Caspase-9 and Apaf-1 bind to each other via their respective NH2-terminal CED-3 homologous domains in the presence of cytochrome c and dATP, an event that leads to caspase-9 activation. Activated caspase-9 in turn cleaves and activates caspase-3. Depletion of caspase-9 from S 100 extracts diminished caspase-3 activation. Mutation of the active site of caspase-9 attenuated the activation of caspase-3 and cellular apoptotic response in vivo, indicating that caspase-9 is the most upstream member of the apoptotic protease cascade that is triggered by cytochrome c and dATP. PMID- 9390558 TI - Regulating the yeast kinetochore by ubiquitin-dependent degradation and Skp1p mediated phosphorylation. AB - In S. cerevisiae, the four-protein Cbf3 complex binds to the essential CDEIII region of centromeric DNA to initiate kinetochore assembly. We report the reconstitution of Cbf3p from recombinant proteins and an analysis of its p58Ctf13 and p23Skp1 subunits. p23Skp1 has both G1- and G2-specific functions in yeast and binds to p58Ctf13 and to the essential Cdc4p component of the ubiquitin conjugating complex Scul(Cdc4). We show that the function of p23Skp1 in Cbf3p is to activate p58Ctf13 by phosphorylation. p58Ctf13 is an unstable protein that is targeted to the proteosome, probably by Scul(Cdc4)-mediated ubiquitination. Thus, p58 appears to be activated by phosphorylation in a p23Skp1-dependent step and degraded by the proteosome in a ubiquitin-dependent step. We propose that coupled activation and destruction link the assembly of Cbf3p to the duplication of centromeres in S phase. PMID- 9390559 TI - Molecular reconstruction of Sleeping Beauty, a Tc1-like transposon from fish, and its transposition in human cells. AB - Members of the Tc1/mariner superfamily of transposons isolated from fish appear to be transpositionally inactive due to the accumulation of mutations. Molecular phylogenetic data were used to construct a synthetic transposon, Sleeping Beauty, which could be identical or equivalent to an ancient element that dispersed in fish genomes in part by horizontal transmission between species. A consensus sequence of a transposase gene of the salmonid subfamily of elements was engineered by eliminating the inactivating mutations. Sleeping Beauty transposase binds to the inverted repeats of salmonid transposons in a substrate-specific manner, and it mediates precise cut-and-paste transposition in fish as well as in mouse and human cells. Sleeping Beauty is an active DNA-transposon system from vertebrates for genetic transformation and insertional mutagenesis. PMID- 9390560 TI - Enteropathogenic E. coli (EPEC) transfers its receptor for intimate adherence into mammalian cells. AB - Enteropathogenic E. coli (EPEC) belongs to a group of bacterial pathogens that induce epithelial cell actin rearrangements resulting in pedestal formation beneath adherent bacteria. This requires the secretion of specific virulence proteins needed for signal transduction and intimate adherence. EPEC interaction induces tyrosine phosphorylation of a protein in the host membrane, Hp90, which is the receptor for the EPEC outer membrane protein, intimin. Hp90-intimin interaction is essential for intimate attachment and pedestal formation. Here, we demonstrate that Hp90 is actually a bacterial protein (Tir). Thus, this bacterial pathogen inserts its own receptor into mammalian cell surfaces, to which it then adheres to trigger additional host signaling events and actin nucleation. It is also tyrosine-phosphorylated upon transfer into the host cell. PMID- 9390561 TI - Identification and molecular characterization of fractalkine receptor CX3CR1, which mediates both leukocyte migration and adhesion. AB - Leukocyte trafficking at the endothelium requires both cellular adhesion molecules and chemotactic factors. Fractalkine, a novel transmembrane molecule with a CX3C-motif chemokine domain atop a mucin stalk, induces both adhesion and migration of leukocytes. Here we identify a seven-transmembrane high-affinity receptor for fractalkine and show that it mediates both the adhesive and migratory functions of fractalkine. The receptor, now termed CX3CR1, requires pertussis toxin-sensitive G protein signaling to induce migration but not to support adhesion, which also occurs without other adhesion molecules but requires the architecture of a chemokine domain atop the mucin stalk. Natural killer cells predominantly express CX3CR1 and respond to fractalkine in both migration and adhesion. Thus, fractalkine and CX3CR1 represent new types of leukocyte trafficking regulators, performing both adhesive and chemotactic functions. PMID- 9390562 TI - Crx, a novel otx-like homeobox gene, shows photoreceptor-specific expression and regulates photoreceptor differentiation. AB - We have isolated a novel otx-like homeobox gene, Crx, from the mouse retina. Crx expression is restricted to developing and mature photoreceptor cells. CRX bound and transactivated the sequence TAATCC/A, which is found upstream of several photoreceptor-specific genes, including the opsin genes from many species. Overexpression of Crx using a retroviral vector increased the frequency of clones containing exclusively rod photoreceptors and reduced the frequency of clones containing amacrine interneurons and Muller glial cells. In addition, presumptive photoreceptor cells expressing a dominant-negative form of CRX failed to form proper photoreceptor outer segments and terminals. Crx is a novel photoreceptor specific transcription factor and plays a crucial role in the differentiation of photoreceptor cells. PMID- 9390564 TI - Enhancement of natural killer and antibody-dependent cytolytic activities of the peripheral blood mononuclear cells of HIV-infected patients by recombinant IL-15. AB - Natural killer (NK) cells are an important subset of lymphocytes capable of killing virus-infected target cells without prior sensitization. HIV-infected individuals show impairment of their NK cell activity. Although the mechanism responsible for this defect remains unclear, NK cytotoxicity of lymphocytes from these individuals can be partially restored by interleukin (IL)-2. IL-15 is a recently discovered cytokine that shares many biologic activities with IL-2--for example, enhancement of NK activity. In this study, we investigated the effect of recombinant IL-15 (rIL-15) on the NK and antibody-dependent cellular cytotoxicity (ADCC) effector activities of peripheral blood mononuclear cells (PBMCs) from HIV infected individuals using K562 cell line and HIV gp120-expressing cells. The effect of anti-IL-15 antibodies on NK activity was also examined using PBMCs of HIV-seronegative individuals. Our results show that NK and ADCC activities of PBMCs in HIV-seropositive patients were significantly lower than those of seronegative donors (p < or = 0.05). However, these two activities were significantly enhanced when rIL-15 was added to the assay wells (p < or = 0.05). Moreover, addition of saturating concentrations of neutralizing monoclonal antibodies (mAb) specific for IL-2, IL-12, or interferon (IFN)-gamma in the assays failed to inhibit IL-15-mediated enhancement of NK cell functions. Only the antibody against IL-15 abrogated the upregulation of NK and ADCC activities mediated by IL-15, suggesting that this cytokine enhances NK cell functions through a mechanism that is independent of the induction of other cytokines. IL 15 did not exert any modulatory effect on the expression of CD16 or CD56 molecules. Our results show that IL-15 can increase the NK and ADCC activities of the PBMCs of HIV-infected individuals in vitro. In view of its higher therapeutic index as determined using murine models, IL-15 may represent a better immunotherapeutic agent than IL-2 to restore these functions in HIV-seropositive patients. PMID- 9390563 TI - Cone-rod dystrophy due to mutations in a novel photoreceptor-specific homeobox gene (CRX) essential for maintenance of the photoreceptor. AB - Genes associated with inherited retinal degeneration have been found to encode proteins required for phototransduction, metabolism, or structural support of photoreceptors. Here we show that mutations in a novel photoreceptor-specific homeodomain transcription factor gene (CRX) cause an autosomal dominant form of cone-rod dystrophy (adCRD) at the CORD2 locus on chromosome 19q13. In affected members of a CORD2-linked family, the highly conserved glutamic acid at the first position of the recognition helix is replaced by alanine (E80A). In another CRD family, a 1 bp deletion (E168 [delta1 bp]) within a novel sequence, the WSP motif, predicts truncation of the C-terminal 132 residues of CRX. Mutations in the CRX gene cause adCRD either by haploinsufficiency or by a dominant negative effect and demonstrate that CRX is essential for the maintenance of mammalian photoreceptors. PMID- 9390565 TI - HIV-1 proviral DNA load across neuroanatomic regions of individuals with evidence for HIV-1-associated dementia. AB - A definitive relation between HIV-1 load and the clinical diagnosis of HIV-1 associated dementia (HAD) has not yet been established. Knowledge of the neuroanatomic distribution of HIV-1 load in the brain of individuals with HAD and HIV-1 encephalitis may facilitate elucidation of this relation. Nine individuals with AIDS were analyzed postmortem by three independent methods with each assessment performed blinded to the others: 1) a neuropsychiatric review of clinical records for evidence of possible HAD, 2) HIV-1 DNA load determination by quantitative polymerase chain reaction (PCR) across several neuroanatomic regions, and 3) a pathologic examination for diagnosis of HIV-1 encephalitis by immunohistochemical techniques. Of eight AIDS cases with clinical records sufficient for neuropsychiatric review, seven were shown to have evidence for HAD. HIV-1 DNA was detected and quantified in specimens from all of the medial temporal lobe regions analyzed but was not detectable in the frontal lobe at the same level of sensitivity in two of these cases (<1 per 1000 cellular genomes). HIV-1 DNA load in the medial temporal lobe region was significantly larger than that in the frontal lobe. Only four of seven cases with evidence for HAD were also diagnosed with HIV-1 encephalitis. PMID- 9390566 TI - Regional differences in use of antiretroviral agents and primary prophylaxis in 3122 European HIV-infected patients. EuroSIDA Study Group. AB - Little is known about how widely HIV-related drugs are used outside controlled clinical trials. We therefore assessed factors associated with use of antiretroviral (ARV) therapy and primary prophylactic regimens to prevent HIV associated opportunistic infections. Baseline data from a prospective study from May to August 1994, on 3122 consecutive HIV infected patients with a CD4 count <500 cells/microl, followed in 37 centers from 16 European countries, were analyzed. Two thousand and twenty patients (65%) were receiving at least 1 ARV drug at the time of the study. ARV therapy was more frequently used among patients from southern and central Europe as compared with patients from northern Europe, especially among patients with CD4 counts >200 cells/microl (73%, 57%, and 42%, respectively, p < 0.0001). Of patients on ARV therapy, 34% received open label combination therapy. This proportion was higher in central Europe compared with other regions (27%, 50%, and 31% for southern, central, and northern Europe, respectively, p < 0.0001). Primary prophylaxis against Pneumocystis carinii pneumonia (PCP) was used by 85% of patients with a CD4 count <200 cells/microl, without marked regional differences. In patients without esophageal candidiasis or other invasive fungal infections, antifungal drugs were far less frequently used in patients from southern and central Europe compared with patients from northern Europe (10%, 10%, and 25%, respectively, p < 0.0001). Only 5% of patients with a CD4 count <100 cells/microl received rifabutine as primary prophylaxis against nontuberculous mycobacterioses. ARV and antifungal therapies are used differently in different parts of Europe, whereas primary PCP prophylaxis is uniformly administered to most at-risk patients. U.S. recommendations on the use of antimycobacterial prophylaxis have not been implemented in Europe. PMID- 9390567 TI - Evaluation of immunologic markers in cervicovaginal fluid of HIV-infected and uninfected women: implications for the immunologic response to HIV in the female genital tract. AB - We analyzed 21 cervicovaginal lavage (CVL) specimens from 19 women participating in the Women's Interagency HIV Study to characterize levels of antibody, cytokine, and complement and to determine associations between these levels and stage of the menstrual cycle, HIV status, and the presence of concurrent genital infection and genital dysplasia. Sixteen samples were collected from HIV-infected women and five from high-risk HIV-seronegative women. CVL fluid was assayed for levels of IgG, secretory IgA (s-IgA), interleukin 2 (IL-2), IL-10, IL-6, tumor necrosis factor alpha (TNF-alpha), IL-1beta, interferon gamma (IFN-gamma), C3, C1q, and C4. Women with HIV were more likely to have cervicovaginal dysplasia (9/16 vs. 0/5; p = 0.027) but were not more likely to have concurrent vaginal infection (10/16 vs. 2/5; p = 0.38). Antibody, cytokine, and complement were detectable in all samples, although not all samples had measurable IL-10, C3, or C4. HIV-infected women demonstrated a trend toward higher levels of IFN-gamma than did uninfected women (p = 0.098); no differences were noted in other parameters. HIV-infected women with vaginal infections had significantly higher CVL levels of IgG (p = 0.023) and IFN-gamma (p = 0.02) than did HIV-infected women without genital infections. HIV-infected women with cervicovaginal dysplasia were found to have higher levels of IL-1beta (p = 0.045) and IFN-gamma (p = 0.039) than those without. Analysis of the HIV-infected cohort by CD4 cell count revealed higher levels of IgG and IFN-gamma in CVL from women with lower CD4 cell counts, although these differences were not statistically significant. Higher levels of proinflammatory cytokines in CVL fluid of women with genital infection or cervicovaginal dysplasia may affect local HIV replication and may influence the risk of acquisition or transmission of HIV for women with these underlying conditions. PMID- 9390568 TI - Are HTLV-II-seropositive injection drug users at increased risk of bacterial pneumonia, abscess, and lymphadenopathy? AB - Disease associations of HTLV-II are poorly defined, despite a high seroprevalence among injection drug users (IDU). One hundred twenty-four HTLV-II-seropositive emergency room and clinic patients were matched by age, sex, and clinic to 120 HTLV-I/II-seronegative patients. Medical records were reviewed blinded to HTLV-II status, and International Classification of Disease 9th Clinical Modification (ICD-9CM)-coded diagnoses were compared between seropositive patients and controls. After adjustment for relevant confounding variables such as human immunodeficiency virus infection, HTLV-II-seropositive IDU had an increased risk of bacterial pneumonia (odds ratio [OR], 3.45; 95% confidence interval [CI], 1.58, 7.56), abscess (OR, 8.30; 95% CI, 4.02, 17.11), and lymphadenopathy (OR, 3.91; 95% CI, 1.24, 12.32) compared with HTLV-II-negative non-IDU patients. In contrast, HTLV-II-negative IDU were at only marginally increased risk of the same conditions, with OR of 1.76 (95% CI, 0.42, 7.40), 3.00 (95% CI, 0.94, 9.59), and 1.31 (95% CI, 0.15, 11.66), respectively. These results indicate that HTLV-II seropositivity may define a subgroup of IDU who are at particularly high risk of bacterial pneumonia, skin and soft tissue abscess, and lymphadenopathy. Whether HTLV-II has an etiologic role in predisposing IDU to bacterial infections and lymphadenopathy will require further investigation. PMID- 9390569 TI - Clostridium difficile-associated diarrhea in HIV-infected patients: epidemiology and risk factors. AB - A retrospective analysis of all the cases of Clostridium difficile-associated diarrhea (CDAD) in hospitalized patients infected with HIV was performed over a 52-month period to assess the incidence, epidemiology, and risk factors of CDAD. A case of CDAD was defined as a patient with diarrhea and a positive stool cytotoxin B assay. Sixty-seven cases of CDAD were recorded in HIV-infected patients between January 1991 and April 1995. The annual incidence of CDAD ranged from 1.7 to 6.4 per 100 HIV-infected patients discharged from hospital. The 67 CDAD cases included 48 (72%) first episodes and 19 (28%) relapses. Serogroup C accounted for 69% of strains from initial episodes of CDAD. To identify risk factors for CDAD, 34 HIV-infected patients with a first episode were compared with 66 HIV-infected controls matched for the length of hospital stay. Three independent factors remained significantly associated with CDAD among HIV infected patients: CD4+ cell counts <50/mm3 (OR = 5.2; 95% CI = 1.4-19.3; p = 0.01), clindamycin use (OR = 5.0; 95% CI = 1.3-18.3; p = 0.02) and penicillin use (OR = 4.6; 95% CI = 1.1-18.8; p = 0.03). C. difficile is a common enteric pathogen responsible for nosocomial diarrhea in HIV-infected patients. Clinicians should keep this pathogen in mind when searching for the cause of diarrhea in these patients, especially those who are severely immunocompromised or have received clindamycin or penicillin. PMID- 9390570 TI - Projected incidence of AIDS in San Francisco: the peak and decline of the epidemic. AB - To predict the incidence of AIDS from 1978 through 1998 in San Francisco, we developed a model that combined annual HIV seroconversion rates for homosexual and bisexual men and for heterosexual injecting drug users with estimates of the incubation period distribution between HIV seroconversion and AIDS diagnosis and with estimates of the size of the at-risk populations. Our model assumed the availability of antiretroviral therapy at the efficacy level of zidovudine monotherapy. The annual number of new AIDS cases is estimated to have peaked at 3332 in 1992, and is projected to decline to 1196 annually by 1998. Although the projected number of cases decreased steadily during this period for homosexual and bisexual men, the projected number of cases for injection drug users, women, and persons with other risks increased between 1993 and 1998. The decline in the incidence of AIDS in San Francisco reflects the dramatic reductions in new HIV infections that occurred a decade previously and that were achieved as a result of significant changes in high-risk behaviors, primarily among homosexual and bisexual men. Changes in HIV seroincidence must be factored in before attributing the decrease in AIDS incidence to more effective combination antiretroviral treatment. PMID- 9390571 TI - A population-based study determining the incidence of tuberculosis attributable to HIV infection. AB - Although the tuberculosis (TB) epidemic has been attributed in part to the AIDS epidemic, few studies in the United States have measured the risk attributable to HIV infection. We linked the TB registry of Alameda County, California, 1985 to 1994, with the AIDS registry, 1982 to 1994. We defined a person with TB and HIV infection as a patient in the TB registry with the same name, race/ethnicity, gender, and date of birth as a patient in the AIDS registry. We used population and HIV seroprevalence estimates to determine the HIV-seropositive and seronegative population at risk of TB in 1994. Of 1990 TB cases reported by Alameda County from 1985 to 1994, 116 (5.8%) had an AIDS diagnosis. Among 25- to 44-year-old TB patients, 25.2% of U.S.-born men and 8.4% of U.S.-born women had an AIDS diagnosis. In 1994, the estimated TB incidence rate in persons with HIV infection was 198.1 per 100,000 versus a rate of 13.9 of 100,000 among persons without HIV infection (rate ratio, 13.8; 95% confidence interval, 8.0, 23.8). In 1994, 93% of TB cases among HIV seropositive persons, 6.4% of all TB cases, and 16.7% of TB cases aged 25 to 44 years were attributable to HIV infection. The high attributable risk underscores the impact of HIV on the TB epidemic. All persons with HIV infection should be screened for TB, and persons with TB infection should be screened for HIV infection. TB/HIV coinfected patients should be provided with TB preventive therapy. PMID- 9390572 TI - Intermediate-size trials for the evaluation of HIV vaccine candidates: a workshop summary. AB - There has been considerable debate over what evidence from preclinical and clinical studies is required to advance an HIV vaccine candidate to phase III efficacy testing. Given this situation, conduct of intermediate-size trials is proposed as a method for assessing the plausibility that a vaccine candidate would prevent chronic HIV infection. Designed to observe 45 incident infections in the control group, these preliminary efficacy trials could rule out candidates with low or no efficacy while advancing those candidates with some evidence of protection to definitive trials. In addition, these trials could provide clues about correlates of immunity. A threefold or greater difference in the postvaccination geometric mean titer of neutralizing antibody can be readily detected between infected and uninfected vaccinees. Differences in CD8+ cytotoxic T lymphocytes, however, are more difficult to detect. Intermediate-size trials could also discern a 0.5 log10 or greater difference in plasma HIV-1 RNA levels between infected vaccinees and infected controls. Such differences in viral load might suggest disease amelioration or reduction in infectiousness. Given the large variability in CD4 count and its relatively modest average decline in the year after infection, a slower decline in CD4 count among infected vaccinees would not be detectable. With limited resources, intermediate-size trials could contribute significantly to HIV vaccine development. PMID- 9390573 TI - Seroprevalence of HIV-1, HIV-2, and HIV-1 group O in Nigeria: evidence for a growing increase of HIV infection. AB - To determine current data on HIV infection and to further confirm the presence of HIV-1 group O infection in Nigeria, 2300 samples from five states were tested for the presence of HIV antibody. A convenience sampling was obtained from pregnant women, tuberculosis (TB) patients, commercial sex workers (CSWs), blood donors, patients with sexually transmitted diseases (STDs), patients with skin diseases, male clients of CSWs, outpatients suspected to have AIDS, truck drivers, and community dwellers. With the exception of pregnant women, the HIV prevalences in all these groups were high: 60.6% in CSWs, 16.2% in TB patients, 7.7% in blood donors in some states, and 16% in the rural area of Kano State. Male clients of CSWs, truck drivers, and STD patients had prevalences of 7.8%, 8.6%, and 21.2%, respectively. Regional differences in relation to HIV prevalences were observed; HIV-2 and most of the HIV-1/2 infections were found in the southern states of Nigeria. Higher HIV prevalences were observed in the north-northeast in pregnant women, TB patients, and CSWs, but for blood donors, higher rates were seen in the southeast-southwest. One asymptomatic 50-year-old woman, a community dweller in Kano, was identified to be HIV-1 group O-positive. Compared with data from national surveillance studies in 1991/1992 and 1993/1994, a substantial increase in HIV infection was observed. Our results show a growing incidence of HIV infection in Nigeria and suggest the presence of a rural HIV epidemic. The identification of HIV-1 group O in Kano shows that this virus strain is geographically widespread in Nigeria. PMID- 9390574 TI - HIV risk profile of drug-using women who have sex with women in 19 United States cities. AB - The objective of this study was to analyze HIV-related risks of women injection drug users (IDU) and crack cocaine users (CCU) who have sex with women (WSW). IDU and CCU women (N = 3856) were recruited from street settings in 19 U.S. cities between 1992 and 1994. For this study, we analyze data on 231 women who reported female sex partners in the 30 days before interview. In the 30 days before interview, 53% of IDUs had shared syringes, and 66% had shared injection supplies. Only 11 women (6%) always used barrier protection while giving oral sex to women and 5 (3%) while receiving oral sex from women in the 30 days before interview. Fifty percent had sex with men as well as women in the previous 30 days. Thirty percent of women who reported sex with men had used condoms for penile-vaginal sex, and 26% for penile-anal sex. In logistic regression analysis modeling sex with men in the previous 30 days, sex work was predictive, "lesbian" self-identification was protective, and the interaction between these two terms was predictive, while controlling for race and age. Differences in risk perception were significant between women who reported varying sexual risks, but not significant between women who reported varying injection-related risks. There is a high prevalence of risky sex and drug behaviors among drug-using WSWs. There is a need for epidemiological studies specifically geared toward studying risk behaviors among WSWs. Risk reduction activities need to focus on injection related risks, as well as sex-related risks, among WSWs. PMID- 9390575 TI - Subtyping HIV-1 by improved resolution of heteroduplexes on agarose gels. PMID- 9390576 TI - Successful use of boric acid to control azole-refractory Candida vaginitis in a woman with AIDS. PMID- 9390577 TI - Subspecialty training in pediatric anesthesiology: what does it mean? PMID- 9390578 TI - Multi-institutional survey of graduates of pediatric anesthesia fellowship: assessment of training and current professional activities. AB - We surveyed all the graduates of four fellowship programs in pediatric anesthesia between 1985 and 1993 to assess their current professional activities, their evaluation of fellowship training, and their opinions on future directions of such training. One-hundred ninety-one (62%) of the graduates responded. Nearly all of the respondents had sought fellowship training for pediatric anesthesia and thought that the training was worthwhile. At the time of the survey, 40% worked in a children's hospital, 72% had university or affiliate positions, and 54% had a practice that was > 50% pediatric. Those with > or = 12 mo fellowship and/or board certification in pediatrics were the most likely to have a pediatric dedicated practice. Seventy percent of the respondents thought that fellowship training should be for 12 mo, and the proportion of respondents who recommended inclusion of training in pain management and clinical research was greater than the number who had actually received such training. Fifty-eight percent of respondents supported restriction of fellowship positions in the future, but 83% did not support a mandatory 2-yr fellowship with research training. We conclude that fellowships in pediatric anesthesia seem to be successful in providing training that is not only satisfying to the trainees, but that is also followed by active involvement in the care of children and in the training of residents and fellows in anesthesia. Additional information should be gathered to assess the impact of this training on pediatric care, to formulate a standardized curriculum, and to justify support for such training in the future. IMPLICATIONS: We surveyed graduates of four fellowship programs in pediatric anesthesia (1985 1993) to assess current professional activities, fellowship training, and future directions of such training. Fellowships in pediatric anesthesia seem to provide training that is satisfying to trainees and that is followed by active involvement in the care of children. PMID- 9390579 TI - Predicting and treating coagulopathies after cardiopulmonary bypass in children. AB - Coagulopathies in children after cardiopulmonary bypass (CPB) are complex. There are very limited data correlating coagulation tests with postoperative bleeding. We evaluated coagulation changes after CPB and after the administration of coagulation products to 75 children. Baseline coagulation tests were obtained and repeated after protamine administration, after transfusion of individual coagulation products, and on arrival in the intensive care unit (ICU). Regression analysis demonstrated no baseline coagulation test to predict postoperative chest tube drainage. Weight and duration of CPB were determined to be the only predictors of bleeding. Further analyses demonstrated that children <8 kg had more bleeding and required more coagulation products than children >8 kg. Postprotamine platelet count and fibrinogen level correlated independently with 24-h chest tube drainage in children <8 kg, whereas postprotamine platelet count and thrombelastographic values did so in patients weighing >8 kg. Platelet administration alone was found to restore effective hemostasis in many patients. With ongoing bleeding, cryoprecipitate improved coagulation parameters and limited blood loss. Fresh-frozen plasma administration after platelets worsened coagulation parameters and was associated with greater chest tube drainage and more coagulation product transfusions in the ICU. Objective data to guide post CPB component therapy transfusion in children are suggested. IMPLICATIONS: Children <8 kg can be expected to have more severe coagulopathies, require more coagulation product transfusions, and bleed more after cardiopulmonary bypass. Correlations between coagulation tests and postoperative chest tube drainage are defined. Platelets and, if necessary, cryoprecipitate optimally restore hemostasis. Fresh-frozen plasma offers no benefits in correcting postcardiopulmonary bypass coagulopathies in children. PMID- 9390580 TI - Sevoflurane or halothane anesthesia: can we tell the difference? AB - This study was performed to evaluate the ability of anesthesiologists to differentiate between sevoflurane, a newer, more expensive anesthetic, and halothane. A total of 113 assessments were made by 36 anesthesiologists on 58 children, aged 6 mo to 6 yr, scheduled for bilateral myringotomy and tube placement. All patients received midazolam (0.5 mg/kg per os) approximately 30 min before the induction of anesthesia. Sevoflurane or halothane was randomly selected for anesthetic induction and maintenance. The anesthesiologists, who were unaware of the anesthetic being used, were asked to identify the anesthetic based on clinical signs and to assess the quality of induction, speed of induction, and speed of emergence using a visual analog scale (VAS; minimum score = 0, maximum score = 100). The anesthesiologists correctly identified the anesthetic only 56.6% of the time. This was not significantly different from the 50% that would result from random guessing (P = 0.08). Further, there were no significant differences in VAS scores between the two groups. This study suggests that in premedicated pediatric patients undergoing brief surgical procedures, anesthesiologists cannot correctly differentiate between sevoflurane and halothane. The lack of significant differences in VAS scores suggests that the speed of induction, the speed of emergence, and the quality of induction are similar under these clinical conditions. Any purported benefits of sevoflurane seem to be of minor consequence under the circumstances studied. IMPLICATIONS: When the anesthetic halothane or sevoflurane is administered in a blind, randomized fashion, anesthesiologists could not reliably identify which drug was being used to anesthetize children for a brief surgical procedure. These results suggest that the differences between the two drugs in clinical practice are small and may not justify the additional cost of sevoflurane. PMID- 9390581 TI - Adverse events and risk factors associated with the sedation of children by nonanesthesiologists. AB - After implementation of hospital-wide monitoring standards, a quality assurance (QA) tool was prospectively completed for 1140 children (aged 2.96 +/- 3.7 yr) sedated for procedures by nonanesthesiologists. The tool captured data regarding demographics, medications used, adequacy of sedation, monitoring, adverse events, and requirement for escalated care. The medical records of children who experienced adverse events were reviewed. Most (99%) children were monitored with pulse oximetry. Chloral hydrate was the most frequently used sedative (74.9% of cases). Of the children, 239 (20.1%) experienced adverse events related to sedation, including inadequate sedation in 150 (13.2%) and decrease in oxygen saturation in 63 (5.5%). Five of these children experienced airway obstruction and two became apneic. No adverse event resulted in long-term sequelae. Of the 854 children who received chloral hydrate, 46 (5.4%) experienced decreased oxygen saturation (> or = 90% of baseline). Children experienced desaturation after the use of chloral hydrate had received the recommended doses of chloral hydrate (38 83 mg/kg). ASA physical status III or IV and age < 1 yr were predictors of increased risk of sedation-related adverse events. These data underscore the importance of appropriate monitoring that includes pulse oximetry to permit early detection of adverse events. IMPLICATIONS: This quality assurance study highlights the risks associated with the sedation of children and emphasizes the importance of appropriate monitoring by trained personnel. Children with underlying medical conditions and those who are very young are at increased risk of adverse events, which indicates that a greater degree of vigilance may be required in these patients. PMID- 9390582 TI - Sinus venosus atrial septal defect? PMID- 9390583 TI - Successful treatment of uncontrollable posttraumatic intracranial hypertension with dihydroergotamin in a child. PMID- 9390584 TI - The effects of epsilon-aminocaproic acid on fibrinolysis and thrombin generation during cardiac surgery. AB - Despite the efficacy of antifibrinolytic drugs in reducing bleeding after cardiac surgery, concerns remain regarding their potential to promote thrombosis. We examined the effect of the antifibrinolytic drug, epsilon-aminocaproic acid (EACA) on fibrinolysis and thrombin generation during cardiac surgery. Forty-one adults undergoing primary coronary artery bypass graft surgery requiring cardiopulmonary bypass (CPB) were prospectively randomized in a double-blind trial to receive either saline or EACA. A loading dose of 150 mg/kg EACA was given before anesthetic induction, followed by a 15 mg x kg(-1) x h(-1) infusion, which continued until 3 h after CPB. Plasma samples for the measurement of D dimer, thrombin-antithrombin III, and soluble fibrin were obtained before surgery, 1 h on CPB, and 3 and 20 h after CPB. In the EACA group, fibrinolytic activity, as measured by D-dimer, was significantly decreased 3 h after CPB, (0.51 +/- 0.15 mg/L vs 1.13 +/- 0.14 mg/L, P < 0.005). Decreased fibrinolytic activity was accompanied by decreased bleeding in the EACA group (660 +/- 127 mL vs 931 +/- 113 mL, P < 0.05). No differences in the generation of thrombin or soluble fibrin were apparent between the two groups. Suppression of fibrinolytic activity in the absence of concomitant reductions in thrombin generation suggests that EACA could potentiate a hypercoagulable prethrombotic state in the perioperative setting. IMPLICATIONS: In a randomized, prospective trial of primary cardiac surgery, we demonstrated that the synthetic antifibrinolytic drug epsilon-aminocaproic acid suppresses fibrinolysis with no effects on thrombin generation. These results suggest the potential for synthetic antifibrinolytic drugs to induce a hypercoagulable prethrombotic state in the perioperative setting. PMID- 9390585 TI - Risk factors for acute postoperative renal failure in thoracic or thoracoabdominal aortic surgery: a prospective study. AB - Acute postoperative renal failure is a common complication of thoracic aorta, thoracoabdominal aorta, or aortic arch surgery. To identify variables associated with acute postoperative renal failure, we prospectively studied 475 consecutive patients undergoing thoracoabdominal aortic surgery over a 12-yr period, including those requiring emergent surgery. One hundred twenty-one (25%) patients developed acute postoperative renal failure, and 39 (8%) required hemodialysis. Using multivariate analysis, acute postoperative renal failure was significantly associated with the following variables: age >50 yr (odds ratio [OR] 2.90 [95% confidence interval 1.52-5.53]), preoperative serum creatinine >120 micromol/L (OR 2.76 [1.70-4.48]), duration of left kidney ischemia >30 min (OR 2.01 [1.27 3.17]), packed red cells administration >5 units (OR 2.04 [1.24-3.37]), and Cell Saver administration >5 units (OR 2.31 [1.34-1.96]). Reimplantation of visceral, renal arteries and the Adamkievicz artery; duration of visceral, spinal, and right kidney ischemia; requirement for fresh frozen plasma; administration of aprotinin; extracorporeal circulation; and procedures with circulatory arrest and profound hypothermia were not predictive of postoperative renal failure. In addition, age >50 yr (OR 5.59 [1.31-23.91]), requirement for packed red blood cells >5 unit (OR 3.91 [1.58-9.67]), and preoperative serum creatinine concentration >120 micromol/L (OR 2.26 [1.13-4.53]) were independent factors for acute renal failure requiring hemodialysis. In conclusion, acute renal failure is often observed after thoracic aortic surgery. Numerous predictive factors must be considered when evaluating the etiology of this complication. IMPLICATIONS: Acute postoperative renal insufficiency is a common complication of thoracic aortic surgery. This study found that age >50 yr, preoperative renal dysfunction, duration of renal ischemia, and amount of blood transfusion are significant predictors of this complication. PMID- 9390586 TI - Epidural analgesia and intravenous patient-controlled analgesia result in similar rates of postoperative myocardial ischemia after aortic surgery. AB - To assess the role of postoperative analgesia on myocardial ischemia after aortic surgery, we compared intravenous patient-controlled analgesia (PCA) with thoracic epidural analgesia (TEA). One hundred twenty-four patients were prospectively randomized to the PCA or TEA group. In the TEA group, a T6-7 or T7-8 epidural catheter was inserted before the induction of general anesthesia. Within 1 h of the end of surgery, analgesia and 24-h two-channel Holter monitoring were begun. Myocardial ischemia was defined as ST segment depression > or = 1 mm, 0.06 s after the J point, and lasting for more than 1 min. In the PCA group, a bolus of morphine, 0.05 mg/kg, was given, followed by 0.02 mg/kg of morphine on demand every 10 min. Bupivacaine 0.125% and fentanyl 10 microg/mL was used in the TEA group. Analgesics were titrated to maintain a visual analog scale score < or = 3. The overall incidence of myocardial ischemia was 18.4%-18.2% for TEA and 18.6% for PCA (P = not significant). There were no differences between the groups in the total duration of ischemia per patient (22.2 +/- 119.8 min for TEA and 20.5 +/- 99 min for PCA) and the number of episodes per patient (0.69 +/- 2.1 for TEA and 1.2 +/- 4.9 for PCA). Twenty-three patients had an adverse cardiac outcome, although there were no differences between the groups. The postoperative pain control was superior with TEA. In these patients undergoing elective aortic surgery, the use of postoperative TEA did not result in a lower incidence of early myocardial ischemia compared with intravenous PCA with morphine, despite better analgesia with TEA. IMPLICATIONS: Postoperative myocardial ischemia is associated with adverse cardiac outcome. Using Holter monitoring after aortic surgery, this study shows that the use of thoracic epidural analgesia with bupivacaine and fentanyl did not result in a lower incidence of myocardial ischemia compared with intravenous patient-controlled analgesia with morphine. PMID- 9390587 TI - Cytokine and lipid mediator blood concentrations after coronary artery surgery. AB - This study investigates whether increased levels of cytokines and lipid mediators may be associated with complications after coronary artery bypass grafting (CABG) with extracorporeal circulation (ECC). Hemodynamic measurements and blood samples were obtained in 32 patients before and after the end of ECC and at the 6th and the 24th postoperative hours. Coagulation and pulmonary and cardiovascular functions were specifically assessed postoperatively at the same time. Patients with cardiovascular dysfunction had higher interleukin 8 (IL-8) levels. Higher platelet-activating factor (PAF) and decreased PAF acetylhydrolase activity (AHA, the enzyme that inactivates PAF) levels were found in patients with moderate cardiovascular dysfunction. Interleukin 6 (IL-6), IL-8, and AHA levels correlated with most hemodynamic parameters and creatine phosphokinase myocardial band levels obtained after surgery. Patients with severe lung injury had lower PAF, 6 keto prostaglandin (Pg)F1alpha, and PgE2 levels and higher thromboxane (Tx) B2 concentrations compared with patients without lung injury. Increased IL6 levels were only associated with moderate lung injury. Impaired hemostasis was associated with higher IL6 levels. AHA, IL-6, and IL-8 seem to be associated with cardiovascular dysfunction. The IL-6 blood levels and the ratio of TxB2/6 keto PgF1alpha blood levels are increased during post-CABG lung injury. These results identify an association between specific post-CABG complications and the systemic inflammatory response. The clinical significance of this association remains to be evaluated. IMPLICATIONS: Patients with pulmonary, cardiovascular, or hemostasis dysfunction after cardiopulmonary bypass demonstrate aberrancies in a variety of cytokines and lipid mediators in arterial blood or plasma. The relationship between these findings and inflammatory response-induced complications remains to be determined. PMID- 9390588 TI - Optimal dose of nicardipine for maintenance of hemodynamic stability after tracheal intubation and skin incision. AB - To determine the optimal dose of nicardipine (N) for maintenance of hemodynamic stability during the postinduction period, we designed a randomized, double blind, placebo-controlled, dose-ranging study using four different doses of N administered after a standardized anesthetic induction sequence. A total of 106 patients were assigned to one of the following treatment groups: saline (control), N 0.5 mg (N0.5), N 1 mg (N1), N 2 mg (N2), and N 4 mg (N4). The study medication was administered intravenously (I.V.) in 2.5 mL of saline over 30 s 2 min before laryngoscopy. Mean arterial pressure (MAP) and heart rate (HR) were recorded at 1-min intervals for 15 min after tracheal intubation and for 5 min after skin incision. After intubation, the peak MAP values differed from the preinduction baseline MAP values by 21% +/- 20%, 9% +/- 12%, 1% +/- 13%, -10% +/- 12%, and -15% +/- 13% (mean +/- SD) in the control, N0.5, N1, N2, and N4 groups, respectively. However, the percent change in the pre- to postintubation MAP values (37% to 47%) was similar in all five groups. The highest postintubation HR values were recorded in the N4 group (P < 0.05 versus the other groups). However, the increases in MAP values after skin incision were the least in the N4 group. In conclusion, N1 I.V., administered 2 min before laryngoscopy provides optimal control of arterial blood pressure during the postinduction period. IMPLICATIONS: Acute increases in blood pressure during anesthesia are undesirable in patients with preexisting cardiovascular diseases. This double-blind study found that the calcium-channel blocker, nicardipine, 1 mg intravenously 2 min before tracheal intubation maintained hemodynamic stability during the intraoperative period. PMID- 9390589 TI - Acute hypovolemia may cause segmental wall motion abnormalities in the absence of myocardial ischemia. AB - New segmental wall motion abnormalities (SWMA) detected by echocardiography are considered sensitive and specific markers of myocardial ischemia. However, we have observed new SWMA during pacing-induced reductions in left ventricular filling, which resolved immediately with cessation of the atrial pacing and simultaneous restoration of filling. Therefore, we designed this study to determine whether acute reduction in filling can induce new SWMA in the absence of ischemia. Institution of cardiopulmonary bypass was used as a clinical model of acute reduction in filling, and a beat-by-beat analysis of left ventricular contraction, filling, blood pressures, and electrocardiogram was performed when the drainage of blood to the cardiopulmonary bypass machine rapidly emptied the heart. Acute reduction in filling induced new SWMA in 4 of 38 study patients. All 4 patients had preexisting abnormalities of left ventricular contraction, but translocation of these preexisting SWMA did not explain the new SWMA, nor did myocardial ischemia. We conclude that acute reduction in left ventricular filling can cause new SWMA in the absence of ischemia. This finding limits the usefulness of new SWMA as a marker of ischemia in the presence of acute reduction in filling, such as that secondary to severe hypovolemia. IMPLICATIONS: This study documented that acute reduction in cardiac filling can be associated with new systolic wall motion abnormalities detected by transesophageal echocardiography in the absence of documented myocardial ischemia. These findings indicate that segmental wall motion may not be a valid marker for ischemia in the setting of acute hypovolemia. PMID- 9390590 TI - Drugs to minimize perioperative blood loss in cardiac surgery: meta-analyses using perioperative blood transfusion as the outcome. The International Study of Peri-operative Transfusion (ISPOT) Investigators. AB - Concern about the side effects of allogeneic red blood cell transfusion has increased interest in methods of minimizing perioperative transfusion. We performed meta-analyses of randomized trials evaluating the efficacy and safety of aprotinin, desmopressin, tranexamic acid, and epsilon-aminocaproic acid in cardiac surgery. All identified randomized trials in cardiac surgery were included in the meta-analyses. The primary outcome was the proportion of patients who received at least one perioperative allogeneic red cell transfusion. Sixty studies were included in the meta-analyses. The largest number of patients (5808) was available for the meta-analysis of aprotinin, which significantly decreased exposure to allogeneic blood (odds ratio [OR] 0.31, 95% confidence interval [CI] 0.25-0.39; P < 0.0001). The efficacy of aprotinin was not significantly different regardless of the type of surgery (primary or reoperation), aspirin use, or reported transfusion threshold. The use of aprotinin was associated with a significant decrease in the need for reoperation because of bleeding (OR 0.44, 95% CI 0.27-0.73; P = 0.001). Desmopressin was not effective, with an OR of 0.98 (95% CI 0.64-1.50; P = 0.92). Tranexamic acid significantly decreased the proportion of patients transfused (OR 0.50, 95% CI 0.34-0.76; P = 0.0009). Epsilon-aminocaproic acid did not have a statistically significant effect on the proportion of patients transfused (OR 0.20, 95% CI 0.04-1.12; P = 0.07). There were not enough patients to exclude a small but clinically important increase in myocardial infarction or other side effects for any of the medications. We conclude that aprotinin and tranexamic acid, but not desmopressin, decrease the number of patients exposed to perioperative allogeneic transfusions in association with cardiac surgery. IMPLICATIONS: Aprotinin, desmopressin, tranexamic acid, and epsilon-aminocaproic acid are used in cardiac surgery in an attempt to decrease the proportion of patients requiring blood transfusion. This meta-analysis of all published randomized trials provides a good estimate of the efficacy of these medications and is useful in guiding clinical practice. We conclude that aprotinin and tranexamic acid, but not desmopressin, decrease the exposure of patients to allogeneic blood transfusion perioperatively in relationship to cardiac surgery. PMID- 9390591 TI - Right heart failure in the setting of hemorrhagic shock after pneumonectomy: successful treatment with percutaneous cardiopulmonary bypass. PMID- 9390592 TI - Phlegmasia cerulea dolens, cerebral venous thrombosis, and fatal pulmonary embolism due to heparin-induced thrombocytopenic thrombosis syndrome. PMID- 9390593 TI - Postoperative ulnar neuropathy associated with prolonged ischemia in the upper extremity during coronary artery bypass surgery. PMID- 9390594 TI - The clinical neuromuscular pharmacology of cisatracurium versus vecuronium during outpatient anesthesia. AB - Neither comparisons of the clinical neuromuscular effects of cisatracurium and vecuronium nor comparative studies of their antagonism by neostigmine have been reported. In addition, the efficacy of administering cisatracurium in divided doses has not been investigated. Accordingly, we applied supramaximal electrical stimuli to the ulnar nerve of 165 ASA physical status I and II patients receiving nitrous oxide/alfentanil/propofol anesthesia. Forty-five patients received cisatracurium 5, 10, or 15 microg/kg, and the evoked response at the adductor pollicis was recorded for 15 min. One hundred-twenty patients received cisatracurium 5, 10, or 15 microg/kg or normal saline placebo followed 5 min later by either cisatracurium 100 microg/kg or vecuronium 100 microg/kg (always after placebo). Time to clinical onset (maximal ablation of single twitch response) was measured. When the evoked response spontaneously recovered to 10% of control height, neostigmine 5, 10, 30, or 50 microg/kg or placebo was administered, and recovery of neuromuscular function was recorded for the next 15 min. The clinical onset of vecuronium without priming (2.8 +/- 0.8 min) (mean +/- SD) was significantly (P < 0.05) faster than the onset of cisatracurium without priming (4.6 +/- 1.4 min). Cisatracurium 5, 10, or 15 microg/kg administered before cisatracurium 100 microg/kg significantly (P < 0.05) accelerated the time to complete ablation of the evoked response (3.9 +/- 0.9, 2.9 +/- 0.8, or 3.0 +/- 0.9 min, respectively) compared with cisatracurium 100 microg/kg without priming. The dose of neostigmine required to achieve 50% assisted recovery of the train-of four ratio at 5 min was significantly (P < 0.05) smaller in patients who received vecuronium (29.1 [17.9-55.3] microg/kg) (mean [95% confidence interval]) compared with those who received cisatracurium (53.7 [31.6-131.5] microg/kg). Given its faster clinical onset and greater sensitivity to antagonism by neostigmine, we conclude that vecuronium may be more suitable than cisatracurium for use in outpatient anesthesia. IMPLICATIONS: We investigated the onset of muscle relaxation using intravenous vecuronium and cisatracurium and assessed the ability of neostigmine to antagonize (reverse) this effect. Our results suggest that vecuronium works faster than cisatracurium and is more sensitive to neostigmine. Vecuronium therefore may be more useful than cisatracurium in outpatient anesthesia. PMID- 9390595 TI - The effects of sevoflurane on cerebral hemodynamics during propofol anesthesia. AB - We investigated the cerebral hemodynamic effects of 0.5 and 1.5 minimum alveolar anesthetic concentration (MAC) sevoflurane during propofol anesthesia in 10 patients undergoing supratentorial tumor resection. All patients received a standardized anesthetic, and their lungs were ventilated with a mixture of air and oxygen to produce mild hypocapnia. Anesthesia was then maintained with a propofol infusion. Muscle relaxation was obtained by infusion of atracurium. A transcranial Doppler probe was used to measure red cell flow velocity in the right middle cerebral artery (Vmca). A right-sided jugular bulb catheter was inserted for sampling of jugular bulb blood. After a 30-min period of stabilization and before the start of surgery, baseline arterial and jugular bulb blood samples were drawn to define the arterial-venous oxygen content difference (AVDO2). Mean arterial pressure and Vmca were recorded. Sevoflurane (0.5 and 1.5 MAC) in oxygen/air was then administered, and all measurements were repeated. Administration of sevoflurane at 0.5 MAC did not change Vmca or AVDO2. Sevoflurane (1.5 MAC) did not change Vmca. There was an approximately 25% reduction in AVDO2 (P < 0.05). This suggests that during propofol anesthesia, although 1.5 MAC sevoflurane does not increase red blood cell velocity, there is a relative increase in flow with respect to metabolism. Administration of large dose sevoflurane may be associated with a degree of luxury perfusion. IMPLICATIONS: We investigated the cerebral hemodynamic effects of sevoflurane in patients undergoing neurosurgery. Small-dose sevoflurane (1%) did not change brain blood flow or oxygen consumption. Large-dose sevoflurane (3%) did not change flow velocity but reduced brain oxygen consumption by 25%. Sevoflurane may provide a degree of luxury perfusion. PMID- 9390596 TI - Intrathecal sufentanil, fentanyl, or placebo added to bupivacaine for cesarean section. AB - We compared the effects of intrathecal sufentanil 2.5 and 5 microg, fentanyl 10 microg, and placebo when administered together with hyperbaric bupivacaine 0.5% 12.5 mg for cesarean section. The study was performed in a randomized, double blind fashion in 80 (20 per group) healthy, full-term parturients presenting for elective cesarean section. Postoperative pain was assessed using the visual analog scale (VAS). Duration of complete analgesia was defined as the time from the intrathecal injection to VAS score > 0. Duration of effective analgesia was defined as the time to VAS score > or = 4. No patient experienced intraoperative pain. Complete analgesia was prolonged in all groups receiving opioids. Effective analgesia was prolonged and the 0- to 6-h intravenous opioid requirements were lower in the groups receiving sufentanil compared with those receiving fentanyl and placebo. The need for intraoperative antiemetic medication was greater in the placebo group. Pruritus was a frequent and dose-related side effect in the groups receiving sufentanil. There were no differences in umbilical cord blood gases or neonatal Apgar scores and neurological and adaptive capacity scores among the groups. In conclusion, the addition of sufentanil or fentanyl improved the quality of subarachnoid block compared with placebo. The duration of action was longer for sufentanil than fentanyl. IMPLICATIONS: Small doses of fentanyl or sufentanil (synthetic opioids) added to bupivacaine (local anesthetic) for spinal anesthesia for cesarean section reduce the need for intraoperative antiemetic medication and increase the duration of analgesia in the early postoperative period compared with placebo. PMID- 9390597 TI - Postoperative analgesic requirement after cesarean section: a comparison of anesthetic induction with ketamine or thiopental. AB - In a randomized, double-blind study, we compared postoperative pain and analgesic requirement in patients who underwent elective cesarean section under general anesthesia induced with thiopental 4 mg/kg (n = 20) or ketamine 1 mg/kg (n = 20). Anesthesia was maintained with nitrous oxide and isoflurane. Postoperative analgesia was provided by patient-controlled analgesia (PCA) using morphine. Median (range) time to first PCA demand was greater in the ketamine group (28 [3 134] min) compared with the thiopental group (20.5 [3-60] min; P = 0.04). Median (range) morphine consumption over 24 h was less in the ketamine group (24.3 [3 41] mg) compared with the thiopental group (35 [4-67] mg; P = 0.017). Visual analog scale pain scores were similar between groups. No patients had recall of intraoperative events or unpleasant dreams. Two patients in the thiopental group and one patient in the ketamine group had pleasant intraoperative dreams. Apgar scores were similar between groups. Median umbilical venous pH was higher (7.33 vs 7.31; P = 0.04) and attributable to lower median umbilical venous Pco2 (5.72 vs 6.14 kPa; P = 0.02) in the ketamine group compared with the thiopental group. Induction of anesthesia for cesarean section using ketamine is associated with a lower postoperative analgesic requirement compared with thiopental. IMPLICATIONS: Patients who had anesthesia for cesarean section induced with ketamine required less analgesic drugs in the first 24 h compared with patients who received thiopental. Ketamine, unlike thiopental, has analgesic properties that may reduce sensitization of pain pathways and extend into the postoperative period. PMID- 9390599 TI - Use of outpatient preoperative evaluation to decrease length of stay for vascular surgery. AB - Interventions that decrease perioperative length of stay can result in considerable cost-savings. This study assesses the impact of same-day admission using outpatient preoperative evaluation on the lengths of stay and hospital costs for patients who underwent carotid end-arterectomy (CEA) or lower extremity revascularization (LER). Patient characteristics and length of stay were compared for two 1-yr periods before and after outpatient preoperative evaluation had been implemented. There were no significant differences before and after the initiation of outpatient preoperative evaluation in the CEA and LER groups in mean age and ASA physical status distributions. The average preoperative length of stay decreased significantly from 7.0 to 1.9 days in the CEA group and from 9.0 to 2.8 days in the LER group. This reduction in the length of stay was associated with a cost-savings of $900 per patient and did not have an adverse effect on patient outcome. We conclude that outpatient preoperative evaluation clinics reduce the cost and length of stay in vascular surgery patients. IMPLICATIONS: We found that outpatient preoperative evaluation and same-day admission were associated with a decrease of 4.5 days in the preoperative length of stay for carotid endarterectomy and lower-extremity revascularization. This was not accompanied by increased mortality and led to hospital cost-savings of approximately $900 per patient. PMID- 9390598 TI - The effects of bupivacaine, L-nitro-L-arginine-methyl ester, and phenylephrine on cardiovascular adaptations to asphyxia in the preterm fetal lamb. AB - The preterm fetal lamb that is exposed to clinically relevant plasma concentrations of lidocaine loses its cardiovascular adaptations to asphyxia, and its condition deteriorates further. Nitric oxide (NO) is an important regulator of vascular tone, and local anesthetics are known to inhibit endothelium dependent vasodilation. The purpose of the present study was to determine whether the adverse effects of lidocaine noted in the preterm fetal lamb also occur with bupivacaine and whether the inhibition of NO results in effects similar to those of bupivacaine. Thirty-two chronically prepared pregnant sheep were studied at 117-119 days' gestation. Maternal and fetal blood pressure, heart rate, and acid base state were evaluated. Fetal organ blood flows were determined using 15 microM diameter dye-labeled microspheres. After a control period, mild to moderate asphyxia (fetal PaO2 15 mm Hg) was induced by partial umbilical cord occlusion and maintained throughout the experiment. Ewes in Group I (n = 13) were given a two-step intravenous infusion of bupivacaine for 180 min. Fetuses in Group II (n = 12) received an intravenous injection of L-nitro-L-arginine-methyl ester (L-NAME) (25 mg/kg), and measurements were taken 10 and 30 min after the injection. A third group (Group III) of fetuses (n = 7) were given an intravenous infusion of phenylephrine to mimic the blood pressure increases noted in L-NAME treated fetuses. At 90 min of stable asphyxia, there was a significant decrease in fetal PaO2 and pHa and an increase in PaCO2 and mean arterial blood pressure. There was also an increase in blood flow to the adrenals, myocardium, and cerebral cortex, whereas blood flow to the placenta decreased. Administration of bupivacaine during asphyxia did not affect the changes in mean arterial blood pressure and acid-base state but did abolish the increases in blood flows to the myocardium and cerebral cortex. Injection of L-NAME to the asphyxiated fetus resulted in an increase in mean arterial blood pressure above the level noted at 90 min of cord occlusion, and an increase in fetal PaO2 toward control levels. This was accompanied by a reduction in organ blood flows to preasphyxia levels. In asphyxiated Group III fetuses, titration of the phenylephrine infusion to achieve blood pressure increases similar to those noted with L-NAME were also associated with an increase in fetal PaO2. These data indicate that bupivacaine abolishes some of the circulatory adaptations to mild to moderate asphyxia induced by partial cord occlusion in the preterm fetal lamb. It is not clear whether these effects of bupivacaine are due to inhibition of NO. IMPLICATIONS: In the preterm fetal lamb, clinically relevant plasma concentrations of bupivacaine achieved by intravenous infusion to the pregnant ewe (80% gestation) abolished some of the fetal cardiovascular adaptations to asphyxia induced by partial umbilical cord occlusion. PMID- 9390600 TI - Transdiscal lumbar sympathetic block: a new technique for a chemical sympathectomy. AB - Genitofemoral neuritis, which occurs when the neurolytic solution spreads into the psoas muscle, is the most common complication after neurolytic lumbar sympathetic block. We developed a transdiscal approach for neurolytic lumbar sympathetic block to reduce the danger of genitofemoral neuritis by making a sympathectomy without penetration of the psoas muscle, through which the genitofemoral nerve passes. We attempted transdiscal lumbar sympathetic block in 14 patients for whom the last previous lumbar sympathetic block performed by using the conventional paravertebral method was unsuccessful. Under fluoroscopic guidance, the needle was inserted transdiscally at L2-3 and/or L3-4 and was advanced until its tip pierced the anterior longitudinal ligament. Radiography and computed tomography revealed that the injected contrast media spread along the anterolateral surface of the vertebral column without any flow into the psoas muscle. Alcohol was injected successfully in all patients. During the 1-mo follow up period, no patients had any symptom of genitofemoral neuritis. Thirteen patients who had been suffering from lower extremity pain achieved partial or complete pain relief. One patient with plantar hyperhidrosis achieved persistent anhidrosis. These results suggest that the transdiscal approach can be a technical option for neurolytic lumbar sympathetic block. IMPLICATIONS: Neurolytic lumbar sympathetic block was performed with the needle advanced through the intervertebral disc. With this technique, the risk of genitofemoral neuritis, the most common complication after neurolytic lumbar sympathetic block, was reduced because the needle does not penetrate the psoas muscle, through which the genitofemoral nerve passes. PMID- 9390601 TI - Late-onset preemptive analgesia associated with preincisional large-dose alfentanil. AB - Few studies using systemic opioids have been adequately designed to demonstrate a preemptive effect. We investigated the preemptive effect of intraoperative large dose intravenous (I.V.) opioids over a 72-h period after lower abdominal surgery. Thirty-eight ASA physical status I or II patients undergoing abdominal hysterectomy were studied in a prospective, randomized, double-blind design. Group PRE received alfentanil 70 microg/kg over 10 min before surgical incision; Group POST received alfentanil 70 microg/kg over 10 min after incision. Patients received no other intraoperative opioid. Pain was treated in the recovery room with 2-mg I.V. boluses of morphine and was subsequently managed via patient controlled analgesia (PCA) using morphine sulfate. Visual analog scale pain scores at rest (VAS-R) and on movement (VAS-M) and PCA morphine consumption were recorded for 72 hours. VAS-M and VAS-R scores did not differ at any point, and morphine consumption was similar in both groups over the initial 48 h. Group PRE used significantly less morphine from 48 to 72 h postoperatively (P < 0.02). We conclude that presurgical incisional (i.e., compared with postincisional) large dose opioid exposure results in a modest, late decrease in postoperative morphine consumption, with no clinical impact on early postoperative pain. Timing of the observed reduction coincides with maximal output of substances implicated in experimental hyperalgesia. IMPLICATIONS: When given before surgical incision, alfentanil, a short-acting narcotic, was associated with a reduction in morphine requirements 48-72 h after surgery. Brief interventions may have a delayed and sustained impact on pain perception, possibly by reducing mechanisms of sensitization. PMID- 9390602 TI - Pharmacokinetics and efficacy of long-term epidural ropivacaine infusion for postoperative analgesia. AB - The aim of this study was to evaluate the pharmacokinetics and efficacy of the new local anesthetic ropivacaine when used for epidural infusion for up to 72 h after major orthopedic surgery. Immediately after surgery, an epidural infusion of ropivacaine 2 mg/mL was begun at a rate of 6 mL/h in 11 patients. The infusion rate was then adjusted according to patient analgesic needs or side effects. Blood samples were taken during and after the infusion to determine total and unbound ropivacaine and alpha1-acid glycoprotein (AAG) concentrations. Patients were assessed regularly for sensory and motor block and pain using a visual analog scale (VAS) score (0-100 mm). Ten patients received 63-72 h of infusion. Total plasma concentrations of ropivacaine and binding protein (AAG) increased during the infusion such that free concentrations plateaued or began to fall over time. VAS values during mobilization were less than 40 mm in 93% of patients. The majority of patients had no measurable motor block once the surgical block had regressed. When epidural ropivacaine was titrated to achieve a stable sensory block, there was a low incidence of motor block, and free plasma ropivacaine levels were well below the toxic range. IMPLICATIONS: The pharmacokinetics of continuous epidural infusions of ropivacaine are described in patients for up to 72 h postoperatively. Clinical efficacy and side effects are also reported. An understanding of the plasma concentrations obtained and modes of elimination during prolonged epidural infusion is important for safe, routine clinical use in postoperative analgesia. PMID- 9390603 TI - Neurotoxicological assessment after intracisternal injection of liposomal bupivacaine in rabbits. AB - Experiments were performed on rabbits randomly assigned to intracisternally receive 0.3 mL of plain bupivacaine 5 mg/mL, liposomal bupivacaine 5 mg/mL, bupivacaine-free liposomes, or isotonic phosphate-buffered saline. Mechanical ventilation was initiated or intravenous dopamine was infused when respiratory depression or hypotension occurred. Seven days after the injection, the whole spinal cord was removed and histopathologic characteristics were studied on transverse sections. All preparations were devoid of phosphatidylcholine hydrolysis or oxidation compounds. Solutions without bupivacaine produced transient irritative signs that required sedation in most rabbits. Despite the similar duration of respiratory depression in groups receiving liposomal or plain bupivacaine, liposomes produced significantly prolonged motor blockade (126 vs 70 min). Correction of hypotension after plain bupivacaine required a longer dopamine infusion and larger doses than after liposomal bupivacaine (28 vs 18 min and 74 vs 47 mg). Necrosis was observed in the cervical area of two rabbits (one in the liposomal bupivacaine group and another in the phosphate buffer group). No demyelinated areas were noted in spinal cord examinations. We conclude that liposomal bupivacaine leads to a less severe sympathetic block and to a prolonged motor block, whereas histologic changes are not significantly different among groups. IMPLICATIONS: Multilamellar liposomes containing bupivacaine administered intracisternally to rabbits produce spinal cord histopathologic changes not significantly different from those observed with plain bupivacaine. Sustained release of bupivacaine from liposomes is suggested by the prolonged motor blockade and the reduced severity of arterial hypotension. Use of these liposomes could prolong the local anesthetic effects of bupivacaine. PMID- 9390604 TI - Modulation of synaptosomal plasma membrane-bound enzyme activity through the perturbation of plasma membrane lipid structure by bupivacaine. AB - We investigated modulations of lipid dynamics and lipid-protein interactions of rat brain synaptosomal plasma membrane (SPM) as one of the possible mechanisms by which the local anesthetic bupivacaine (BPV) has an adverse effect on nerve cell function, with SPM-bound enzyme activity used as a functional probe. The kinetics of BPV impact on the activity of the endoenzymes Ca2+/Mg2+-stimulated ATPase and Na+/K+-stimulated ATPase and the active concentrations of the drug were relevant to those that produce biphasic systemic toxicity. Arrhenius plots of these enzymes showed a transition temperature of 26.6 +/- 1.8 degrees C and 24.5 +/- 1.2 degrees C (mean +/- SD), respectively, in control SPM, which shifted to 17.1 +/- 0.95 degrees C (P < 0.01) and 18.2 +/- 0.85 degrees C (P < 0.05) in SPM treated with 10(-5) M BPV. The Hill coefficients for the allosteric inhibition of Ca2+/Mg2+-stimulated ATPase by Na+ and Na+/K+-stimulated ATPase by fluoride decreased from 1.73 +/- 0.20 and 1.95 +/- 0.25, respectively, in controls to 0.92 +/- 0.09 (P < 0.001) and 1.09 +/- 0.11 (P < 0.001) in the presence of 10(-5) M BPV. The fluidity perturbation in the microenvironment of the ectoenzyme acetylcholinesterase was observed only at 5 x 10(-3) M BPV, as confirmed by the disparity in transition temperature between the controls (22.3 +/- 1.2 degrees C) and the BPV-treated SPM (17.5 +/- 0.8 degrees C, P < 0.01) and that in the Hill coefficient in the two groups: 2.15 +/- 0.24 and 0.97 +/- 0.12 (P < 0.001), respectively. IMPLICATIONS: We propose that under physiological conditions, the neutral and protonated forms of local anesthetics can affect nerve cell function through the asymmetric perturbation of the membrane lipid structure, accompanied by synaptosomal plasma membrane-bound enzyme dysfunction. PMID- 9390605 TI - An osteoid osteoma as an undiagnosed cause of three years of severe pain. PMID- 9390606 TI - Safety steps for epidural injection of local anesthetics: review of the literature and recommendations. PMID- 9390607 TI - Spinal anesthesia, hypothermia, and sedation: a case of resedation with forced air warming. PMID- 9390608 TI - Reversal of neuromuscular blockade with neostigmine has no effect on the incidence or severity of postoperative nausea and vomiting. AB - We performed this randomized, double-blind, placebo-controlled study to determine whether reversal of neuromuscular block with neostigmine increases the incidence and severity of postoperative nausea and vomiting (PONV). We studied 162 women undergoing abdominal hysterectomy and randomly allocated them into two groups. In Group A, neuromuscular block produced with mivacurium was antagonized with neostigmine 2.0 mg and glycopyrrolate 0.4 mg intravenously, whereas Group B received no drugs to facilitate antagonism of blockade. The incidence and severity of PONV was assessed up to 27 h after the operation. There was no difference in PONV between the groups (in Group A 35% had nausea and 33% vomited; in Group B 28% nauseated and 40% vomited) or in the amount of antiemetics given. We had a 75% chance to find a 30% difference in PONV. We conclude that the administration of neostigmine and glycopyrrolate at the end of anesthesia to reverse neuromuscular block does not increase the incidence or severity of PONV. IMPLICATIONS: Neostigmine may increase postoperative nausea and vomiting. In this study, omission of reversal of neuromuscular block with neostigmine failed to decrease the incidence or severity of postoperative nausea and vomiting. PMID- 9390609 TI - Time-dependent changes in heart rate and pupil size during desflurane or sevoflurane anesthesia. AB - To better characterize alterations in autonomic function associated with prolonged anesthesia, we tested the hypothesis that the time-dependent effects of sevoflurane and desflurane differ. We studied seven male volunteers, each anesthetized for 8 h with 1.25 minimum alveolar anesthetic concentration desflurane on one study day and with 8 h sevoflurane on another. These volunteers did not undergo surgery and were minimally stimulated during the study. Measurements included blood pressure, heart rate, pupillary size and light reactivity, concentrations of serum catecholamines, and carbon dioxide production. Over time, heart rate and pupil size increased significantly. During 6 of the 14 anesthetics (45%), heart rate at some point exceeded 95 bpm; similarly, pupil size at some time exceeded 5 mm during 8 anesthetics (57%). In contrast, plasma catecholamine concentrations and carbon dioxide production remained unchanged, and blood pressure remained nearly constant. There are thus substantial time-dependent changes in autonomic functions during prolonged anesthesia, even in unstimulated, nonsurgical volunteers, but we could not detect a difference in these changes during desflurane compared with sevoflurane anesthesia. IMPLICATIONS: Pupil size and heart rate changes are used to guide the delivery of anesthesia. In volunteers, pupil size and heart rate increased with increasing duration of constant desflurane or sevoflurane anesthesia. Thus, anesthetic duration alters heart rate and pupil size independent of surgery and changes in anesthetic delivery. PMID- 9390610 TI - Argon pneumoperitoneum is more dangerous than CO2 pneumoperitoneum during venous gas embolism. AB - We investigated the possibility of using argon, an inert gas, as a replacement for carbon dioxide (CO2). The tolerance of argon pneumoperitoneum was compared with that of CO2 pneumoperitoneum. Twenty pigs were anesthetized with enflurane 1.5%. Argon (n = 11) or CO2 (n = 9) pneumoperitoneum was created at 15 mm Hg over 20 min, and serial intravenous injections of each gas (ranging from 0.1 to 20 mL/kg) were made. Cardiorespiratory variables were measured. Transesophageal Doppler and capnographic monitoring were assessed in the detection of embolism. During argon pneumoperitoneum, there was no significant change from baseline in arterial pressure and pulmonary excretion of CO2, mean systemic arterial pressure (MAP), mean pulmonary artery pressure (PAP), or systemic and pulmonary vascular resistances, whereas CO2 pneumoperitoneum significantly increased these values (P < 0.05). During the embolic trial and from gas volumes of 2 and 0.2 mL/kg, the decrease in MAP and the increase in PAP were significantly higher with argon than with CO2 (P < 0.05). In contrast to CO2, argon pneumoperitoneum was not associated with significant changes in cardiorespiratory functions. However, argon embolism seems to be more deleterious than CO2 embolism. The possibility of using argon pneumoperitoneum during laparoscopy remains uncertain. IMPLICATIONS: Laparoscopic surgery requires insufflation of gas into the peritoneal cavity. We compared the hemodynamic effects of argon, an inert gas, and carbon dioxide in a pig model of laparoscopic surgery. We conclude that argon carries a high risk factor in the case of an accidental gas embolism. PMID- 9390611 TI - Carbon dioxide spirogram (but not capnogram) detects leaking inspiratory valve in a circle circuit. AB - Expiratory valve incompetence in the circle circuit is diagnosed by using capnography (PCO2 versus time) when significant CO2 is present throughout inspiration. However, inspiratory valve incompetence will allow CO2-containing expirate to reverse flow into the inspiratory limb. CO2 rebreathing occurs early during the next inspiration, generating a short extension of the alveolar plateau and decreased inspiratory downslope of the capnogram, which may be indistinguishable from normal. We hypothesized that CO2 spirography (PCO2 versus volume) would correctly measure inspired CO2 volume (VCO2) during inspiratory valve leak. Accordingly, a metabolic chamber (alcohol combustion) was connected to a lung simulator, which was mechanically ventilated through a standard anesthesia circle circuit. By multiplying and integrating airway flow and PCO2, overall, expired, and inspired VCO2 (VCO2,br = VCO2,E - VCO2,I) were measured. When the inspiratory valve was compromised (by placing a wire between the valve seat and diaphragm), VCO2,I increased from 2.7 +/- 1.7 to 5.7 +/- 0.2 mL (P < 0.05), as measured by using CO2 spirography. In contrast, the capnogram demonstrated only an imperceptible lengthening of the alveolar plateau and did not measure VCO2,I. To maintain effective alveolar ventilation and CO2 elimination, increased VCO2,I requires a larger tidal volume, which could result in pulmonary barotrauma, decreased cardiac output, and increased intracranial pressure. IMPLICATIONS: Circle circuit inspiratory valve leak will allow CO2 containing expirate to reverse flow into the inspiratory limb, with subsequent rebreathing during the next inspiration. This CO2 rebreathing causes imperceptible lengthening of the alveolar plateau of the capnogram and is detected only by using the CO2 spirogram (PCO2 versus volume). PMID- 9390612 TI - The UpsherScope in routine and difficult airway management: a randomized, controlled clinical trial. AB - The UpsherScope, a rigid fiberoptic laryngoscope, may facilitate tracheal intubation. We performed a randomized, controlled trial of tracheal intubation using the UpsherScope and compared the success rate with that of direct laryngoscopy. Three hundred patients were randomly assigned to either fiberoptic oral intubation using the UpsherScope (Group US, n = 148) or to direct laryngoscopy (Group DL, n = 152). No significant differences in airway variables were observed between the groups. US intubation was successful in 129 of 148 patients (87%). A second or third attempt was required in 15% and 3%, respectively, of the patients successfully intubated with US. The remaining patients were intubated using DL (n = 17) or the flexible fiberoptic bronchoscope (n = 2). The success rate of DL was significantly higher (97%; P < 0.05), with a second or third attempt required in only seven patients. Time needed to perform successful intubation was 50 +/- 41 s for the US group compared with 23 +/- 13 s for the DL group (P < 0.05). We found no advantage of the UpsherScope over direct laryngoscopy during routine and difficult airway management. Time needed, number of attempts required to perform intubation, and incidence of failure were significantly longer and higher in group US. IMPLICATIONS: We studied tracheal intubation using the fiberoptic UpsherScope and compared the success rate with that of a control group of patients intubated using conventional laryngoscopy. No advantages of the new device were found. On the contrary, time needed, number of attempts required, and incidence of failure were even longer and higher. PMID- 9390613 TI - Dehydration of Baralyme increases compound A resulting from sevoflurane degradation in a standard anesthetic circuit used to anesthetize swine. AB - In a model anesthetic circuit, dehydration of Baralyme brand carbon dioxide absorbent increases degradation of sevoflurane to CF2=C(CF3)OCH2F, a nephrotoxic vinyl ether called Compound A. In the present study, we quantified this increase using "conditioned" Baralyme in a circle absorbent system to deliver sevoflurane anesthesia to swine. Mimicking continuing oxygen delivery for 2 days after completion of an anesthetic, we directed a conditioning fresh gas flow of 5 L/min retrograde through fresh absorbent in situ in a standard absorbent system for 40 h. The conditioned absorbent was subsequently used (without mixing of the granules) in a standard anesthetic circuit to deliver sevoflurane to swine weighing 78 +/- 2 kg. The initial inflow rate of fresh gas flow was set at 10 L/min with the vaporizer at 8% to achieve the target end-tidal concentration of 3.0%-3.2% sevoflurane in approximately 20 min. The flow was later decreased to 2 L/min, and the vaporizer concentration was decreased to sustain the 3.0%-3.2% value for a total of 2 h (three pigs) or 4 h (eight pigs). Inspired Compound A increased over the first 30 +/- 60 min to a peak concentration of 357 +/- 49 ppm (mean +/- SD), slowly decreasing thereafter to 74 +/- 6 ppm at 4 h. The average concentration over 2 h was 208 +/- 25 ppm, and the average concentration over 4 h was 153 +/- 19 ppm. Pigs were killed 1 or 4 days after anesthesia. The kidneys from pigs anesthetized for both 2 h and 4 h showed mild inflammation but little or no tubular necrosis. These results suggest that dehydration of Baralyme may produce concentrations of Compound A that would have nephrotoxic effects in humans in a shorter time than would be the case with normally hydrated Baralyme. IMPLICATIONS: The vapor known as Compound A can injure the kidney. Dehydration of Baralyme, a standard absorbent of carbon dioxide in inhaled anesthetic delivery systems, can cause a 5- to 10-fold increase in Compound A concentrations produced from the inhaled anesthetic, sevoflurane, given at anesthetizing concentrations in a conventional anesthetic system. PMID- 9390614 TI - Contribution of the spinal cord to arousal from inhaled anesthesia: comparison of epidural and intravenous fentanyl on awakening concentration of isoflurane. AB - To investigate the contribution of modulation of afferent nociceptive inputs by an opioid in the spinal cord to arousal from inhaled anesthesia, we determined the awakening concentration of isoflurane in 50 unpremedicated patients scheduled for abdominal hysterectomy. Patients were assigned randomly to three groups. Group I received bolus injections of both epidural and intravenous (I.V.) saline, followed by both epidural and I.V. infusions at the rate of 0.2 mL x kg(-1) h( 1). Group II received an I.V. injection of fentanyl 2 microg/kg, followed by an infusion at the rate of 25 ng x kg(-1) x min(-1), and Group III received an epidural injection and infusion in the same administration regimen as Group II. Anesthesia was induced with and maintained by isoflurane in an air/oxygen mixture (fraction of inspired oxygen = 0.5) with no adjuvant drugs. The study drug was administered at the start of retroperitoneal suturing. The awakening concentrations of isoflurane in Groups I, II, and III (mean +/- SD) were 0.32% +/ 0.07%, 0.31% +/- 0.06%, and 0.24% +/- 0.06%, respectively. At that time, plasma fentanyl concentrations in Groups II and III were 1.12 +/- 0.09 ng/mL and 0.65 +/ 0.04 ng/mL, respectively. Epidural fentanyl infusion reduced the awakening concentration of isoflurane more (P < 0.01) than I.V. fentanyl infusion, despite the lower plasma concentration (P < 0.01) in the epidural group. These findings suggest that epidural fentanyl delays arousal from inhaled anesthesia by modulating the afferent nociceptive inputs in the spinal cord. The spinal cord may contribute to arousal from inhaled anesthesia through the regulation of afferent inputs by opioids along with the supraspinal region of the central nervous system (CNS), even if the effects of subarachnoid fentanyl on the higher CNS via the cephalad migration is taken into consideration. IMPLICATIONS: The present study revealed that the spinal cord, the lower level of central nervous system, contributed to arousal from general anesthesia, along with the higher central nervous system, by comparing the concentrations of an inhaled anesthetic, isoflurane, in the expiration of patients receiving systemic or regional administration of an opioid, fentanyl. PMID- 9390615 TI - The choice of anesthetic maintenance technique influences the antiinflammatory cytokine response to abdominal surgery. AB - Outcome in some diseases is determined by the relationship between pro- and antiinflammatory cytokines. Surgery may also provoke a cytokine response, which has both pro- and antiinflammatory components. The aim of this study was to ascertain whether anesthetic technique can modify the balance of cytokines associated with abdominal surgery. Twenty patients scheduled to undergo elective abdominal hysterectomy were randomly allocated to receive maintenance of anesthesia with isoflurane (IH group) or propofol (IV group). Venous blood samples for measurement of tumor necrosis factor alpha (TNF-alpha), interleukin-6 (IL-6), interleukin-10 (IL-10), and interleukin-1 receptor antagonist (IL-1ra) were taken before the induction of anesthesia and at set intervals until 24 h postoperatively. TNF-alpha levels remained low throughout the study; however, all patients showed a significant postoperative increase in IL-6, IL-10, and IL-1ra (P < 0.05). Levels of the proinflammatory cytokine IL-6 were similar in both groups, whereas the antiinflammatory cytokine IL-10 was higher in the IV group at 4 h postoperatively (P < 0.02). The difference between groups in terms of IL-1ra production just failed to reach significance (P < 0.06). We conclude that the cytokine response to abdominal surgery has both pro- and antiinflammatory components and that the choice of anesthetic may modify the balance of these cytokines. IMPLICATIONS: This study demonstrates that in addition to the widely reported proinflammatory cytokine response, elective abdominal surgery provokes an antiinflammatory response, which may be enhanced by total intravenous anesthesia. The ability of anesthetics to modify the cytokine response to surgery may have therapeutic potential. PMID- 9390616 TI - Pharmacokinetics and pharmacodynamics of propofol in a new solvent. AB - Pain on injection is the most commonly reported adverse event after propofol injection. In a randomized, cross-over study in two groups of 12 healthy male volunteers (24-42 yr), we compared the pharmacokinetics and pharmacodynamics of two new propofol formulations (1% and 2% concentrations) in a fat emulsion consisting of medium- and long-chain triglycerides with the standard propofol formulation. After a single intravenous bolus injection of 2 mg/kg, propofol blood levels were measured for 24 h and evaluated according to an open three compartment model. The derived pharmacokinetic variables were not different among formulations. Additionally, electroencephalographic recordings of the onset and duration of hypnotic action were comparable with all formulations. After propofol 1% in the new formulation, fewer volunteers reported severe or moderate pain on injection (9%) than after the standard formulation (59%) (P < 0.05). We attribute this result to a lower concentration of free propofol in the aqueous phase of the new formulation. IMPLICATIONS: Changing the composition of the carrier fat emulsion for propofol does not have an impact on the pharmacokinetics and efficacy of propofol, but it promises to provide better patient acceptance by lowering the incidence of moderate and severe pain on injection. PMID- 9390617 TI - Anesthetic considerations in patients presenting with mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS) syndrome. PMID- 9390618 TI - A case of nonoliguric renal failure after general anesthesia with sevoflurane and desflurane. PMID- 9390619 TI - Cremasteric reflex for identification of successful spinal anesthesia. PMID- 9390620 TI - Use of gabapentin in a case of facial neuritis. PMID- 9390621 TI - Detection of intraoperative segmental wall motion abnormalities by transesophageal echocardiography. PMID- 9390622 TI - Adequate humidification? PMID- 9390623 TI - Preoperative acute normovolemic hemodilution is an alternative to hypervolemic hemodilution--in case of proper use. PMID- 9390624 TI - Device designed to facilitate endotracheal intubation in the absence of a trained assistant (the anesthesiologist's third hand) PMID- 9390625 TI - Pulse oximeter probe sheath for buccal use. PMID- 9390626 TI - Activation of apnea alarm by a surgical theater light during ophthalmological surgery. PMID- 9390627 TI - Light-guided tracheal intubation through the laryngeal mask airway. PMID- 9390628 TI - Resolution of primary vaginismus and introital hyperesthesia by topical anesthesia. PMID- 9390629 TI - Pneumothorax and "mini" thoracotomy. PMID- 9390630 TI - Respiratory depression with addition of fentanyl to spinal anesthesia. PMID- 9390631 TI - Transforming growth factor-beta, other growth factors, and the extracellular matrix. PMID- 9390632 TI - Renal cell proliferation and the two faces of cyclic adenosine monophosphate. PMID- 9390633 TI - Genetic and antigenic variation in Pneumocystis carinii organisms: tools for examining the epidemiology and pathogenesis of infection. PMID- 9390634 TI - Regulation of the human immune response during tuberculosis. AB - Pulmonary tuberculosis is characterized by depression of purified protein derivative-stimulated (PPD-stimulated) blastogenesis in peripheral blood mononuclear cells (PBMCs) as well as decreased production of interleukin-2 (IL-2) and interferon-gamma (IFN-gamma). Circulating T cells and monocytes (MNs) are nonspecifically activated in situ. PPD directly stimulates the primed MNs from patients with tuberculosis (TB) to overproduce a panoply of cytokines including transforming growth factor-beta (TGF-beta) and IL-10, which serve to depress PPD stimulated blastogenesis and cytokine expression. Cross-modulation by these immunosuppressive MN products is superimposed on a primary T cell abnormality that persists for at least 12 months after the diagnosis of TB and involves apoptotic mechanisms. PMID- 9390635 TI - Modulation of collagen gene expression by cytokines: stimulatory effect of transforming growth factor-beta1, with divergent effects of epidermal growth factor and tumor necrosis factor-alpha on collagen type I and collagen type IV. AB - Transforming growth factor-beta1 (TGF-beta1) is well recognized as a potent mediator of both fibrillar (collagen type I) and basement membrane (collagen type IV) production. However, tissue injury is characterized by the concomitant expression of many cytokines and/or growth factors in addition to TGF-beta1, and the ultimate extent of extracellular-matrix (ECM) deposition may reflect the interacting effects of TGF-beta1 and these other cytokines and/or growth factors. We, therefore, sought to determine whether other cytokines and/or growth factors, known to be produced after tissue injury, are capable either alone or in combination with TGF-beta1 of modulating collagen gene expression. Collagen type I and collagen type IV gene expression was assessed in NIH-3T3 cells, a murine fibroblast-like cell line that responds to TGF-beta1, with increases in both collagen type I and collagen type IV production. TGF-beta1 coordinately induced production of collagen type IV messenger ribonucleic acid (mRNA) to a level 3.8 fold above its baseline value (p < 0.001) and collagen type I mRNA to a level 2.6 fold above its baseline value (p < 0.001). Of the other cytokines and/or growth factors tested, only epidermal growth factor (EGF) had significant effects on collagen mRNA expression. We report the novel observation that EGF significantly induced collagen type IV mRNA (3.0-fold; p < 0.001) but did not alter collagen type I mRNA expression. Platelet-derived growth factor (PDGF), basic fibroblast growth factor (bFGF), tumor necrosis factor-alpha (TNF-alpha), interleukin-1 (IL 1), and insulin-like growth factor-1 (IGF-1) did not alter the expression of mRNA for collagen type IV or collagen type I. Addition of TGF-beta1 to cytokine- and/or growth factor-treated cells increased both collagen type IV and collagen type I mRNA levels. However, collagen type IV mRNA levels were similar in cultures given TGF-beta1 alone and cultures given TGF-beta1 with other cytokines and/or growth factors; there were no additive, synergistic, or antagonistic effects after coadministration of TGF-beta1 and other cytokines and/or growth factors. With regard to collagen type I mRNA expression, all cytokines and/or growth factors tested, with the exception of TNF-alpha, had no effect on collagen type I mRNA levels in TGF-beta1-treated cultures. Importantly, TNF-alpha antagonized the stimulatory effect of TGF-beta1 on collagen type I mRNA levels. These observations support a dominant role for TGF-beta1 in stimulating coordinate expression of collagen type I and collagen type IV mRNAs by NIH-3T3 cells; EGF and TNF-alpha are capable of inducing divergent expression of the genes for these two types of collagen. PMID- 9390637 TI - Aspirin treatment of the low-dose-endotoxin-treated pregnant rat: pathophysiologic and immunohistologic aspects. AB - In the present study, we evaluated the effect of low-dose aspirin (acetylsalicylic acid (ASA); 1.0 mg/kg daily) on blood pressure, albumin excretion, glomerular fibrinogen deposits, and glomerular (basement) membrane bound adenosine diphosphatase (ecto-ADPase) activity, as well as on glomerular inflammation in pregnant rats infused with low-dose endotoxin (1.0 mg/kg). Rats (day 14 of pregnancy) were infused with endotoxin (ET rats) or saline (control rats) and received ASA in their drinking water. These rats were compared with non ASA-treated rats. Blood pressure and albumin excretion were measured from day 15 to day 21, and glomerular fibrinogen and ecto-ADPase activity were measured at day 21. Glomerular inflammation was evaluated at various times after the start of the infusion. The results show that treatment with ASA had a significant beneficial effect on hypertension and inflammation induced by endotoxin in pregnant rats, whereas it reduced albumin excretion and glomerular fibrinogen deposits in some of the rats. PMID- 9390636 TI - Inhibitors of cyclic nucleotide phosphodiesterase isozymes block renal tubular cell proliferation induced by folic acid. AB - In previous studies we observed that inhibition of cyclic 3',5'-nucleotide phosphodiesterase (PDE) isozymes, namely isozyme PDE3, suppresses proliferation of rat renal glomerular mesangial cells in vitro and in vivo. To determine whether activation of the cyclic adenosine monophosphate (cAMP)-protein kinase A (PKA) signaling pathway coupled to specific PDE isozymes modulates accelerated proliferation of renal epithelial cells, we investigated the effect of selective PDE isozyme inhibition on renal epithelial cell proliferation induced in rats by injection of folic acid (FA). In extracts from suspensions of renal cortical tubules, cAMP was metabolized predominantly by isozyme PDE4; activity of PDE3 was about three times lower. The increase in proliferative activity of renal cortical tissue from FA-injected rats, evaluated by immunostaining with Mib-1 antibody, was limited to tubular epithelial cells. Administration of the PDE3 inhibitors cilostazol or cilostamide together with the PDE4 inhibitor rolipram blocked mitogenic synthesis of DNA, as determined by (3H)-thymidine incorporation into renal cortical DNA, in FA-treated rats. FA injection caused an increase of more than 10-fold in proliferating cell nuclear antigen (PCNA) in renal cortical tissue; administration of the potent PDE3 inhibitor lixazinone or, to a lesser degree, cilostazol suppressed these high PCNA levels, whereas rolipram alone had no effect. The results indicate that FA-stimulated in vivo proliferation of renal tubular epithelial cells is down-regulated by activation of a cAMP-PKA signaling pathway linked to PDE3 isozymes. These observations are consistent with the notion that negative crosstalk between cAMP signaling and mitogen-stimulated signaling pathways regulates mitogenesis of renal cells of different terminal differentiation, including tubular epithelial cells. PMID- 9390638 TI - Evaluation of the hepatic iron index as a diagnostic criterion for genetic hemochromatosis. AB - The hepatic iron index was originally described as a useful test to discriminate genetic hemochromatosis from alcoholic siderosis. To evaluate the hepatic iron index as a diagnostic criterion, it is essential to evaluate a cohort of hemochromatosis patients in whom the diagnosis has been established with great certainty. The presence of a sibling with identical human leukocyte antigen (HLA) and with iron overload was considered to be the gold standard for the diagnosis. Hepatic iron index was reviewed retrospectively in 55 homozygotes and in 189 patients who did not have hemochromatosis and were referred for hepatic iron analysis. Four of 55 homozygotes (7%) had a hepatic iron index of < or = 1.9. Hepatic iron concentration was increased in all 4 patients, ranging from 36 to 100 micromol/gm dry weight, (normal value <35.5 micromol/gm). Twelve of 189 (6%) patients without hemochromatosis had hepatic iron indexes > 1.9. The positive likelihood ratio for a hepatic iron index of 1.9 was 12.4. Area under the receiver operating characteristic curve was 0.94 (0.9 to 0.99, 95% confidence interval). The hepatic iron index remains a useful tool in the diagnosis of genetic hemochromatosis. However, it should not be an absolute criterion for the diagnosis and should be interpreted in combination with clinical assessment and genetic studies. PMID- 9390639 TI - Viridans streptococcal isolates from patients with septic shock induce tumor necrosis factor-alpha production by murine macrophages. AB - Viridans streptococci are an important cause of bacteremia and septic shock in neutropenic patients, especially patients receiving chemotherapeutic agents that induce severe mucositis. The mechanisms by which viridans streptococci cause septic shock are unclear. We hypothesized that septic shock due to viridans streptococci is attributable to host cytokine production. Three clinical isolates of viridans streptococci were evaluated for their ability to induce production of tumor necrosis factor-alpha (TNF-alpha) by RAW 264.7 murine macrophages. These three strains of viridans streptococci induced TNF-alpha in a dose-dependent fashion, and the kinetics of TNF-alpha induction were similar to those observed with a clinical isolate of Escherichia coli. PMID- 9390640 TI - Energy-dependent expression of platelet-von Willebrand factor on the surface of unstimulated and stimulated platelets. AB - We have studied the energy requirements (adenosine triphosphate) for the expression of platelet-von Willbrand factor on platelets under conditions in which glycolysis and/or oxidative phosphorylation were inhibited. We found that platelet-vWf expression on the surfaces of both unstimulated and stimulated platelets required energy and was maximally decreased when metabolic ATP was maximally depleted. Platelet-vWf expression correlated directly with estimates of adenylate energy charge in both unstimulated and stimulated platelets. In addition, platelet shape change and agonist-induced intracellular Ca2+ flux were maintained at lower AECs than were either platelet aggregation or alpha-granule secretion. Our results indicate that the surface expression of platelet-vWf on unstimulated platelets is a dynamic process, and that energy is required to maintain basal amounts of platelet-vWf on the platelet surface. Our data also suggest that the metabolic ATP required to effect changes in platelet shape is less than that necessary to maintain basal platelet-vWf surface expression or to produce full alpha-granule secretion. We show that platelet-shape change in the absence of alpha-granule secretion can result in an increase in platelet-vWf surface expression. PMID- 9390641 TI - Inhibition of proliferation and induction of apoptosis by doxycycline in cultured human osteosarcoma cells. AB - Matrix metalloproteinases (MMPs) play a major role in the phenomena of growth, invasion, and metastasis of malignant disease. We studied the effects of doxycycline, a synthetic tetracycline that has been shown to suppress MMP activity in other solid tumors, on osteosarcoma (OSA) cell proliferation and MMP activity in vitro. OSA cells from 6 patients and from one established human tumor cell line (U2OS) (American Type Culture Collection) were cultured in the presence or absence of doxycycline. Doxycycline (10 microg/ml) suppressed OSA cell proliferation threefold to sevenfold in all cultures. MMP activity was assessed by gelatin zymography and was diminished by approximately 50% in all cultures. We examined the hypothesis that induction of apoptosis is one of the mechanisms by which doxycycline inhibits OSA cell proliferation. Ethidium bromide-stained gels of DNA from cells grown in the presence of 5 microg/ml and 10 microg/ml of doxycycline revealed laddering consistent with apoptosis after 24 hours in culture. The demonstration that doxycycline suppresses cell proliferation and MMP activity and induces apoptosis in human OSA cells in vitro suggests that this well-tolerated oral agent may be effective in the in vivo treatment of OSA. PMID- 9390642 TI - Proinflammatory cytokines: indicators of infection in high-risk patients. AB - Proinflammatory cytokines play an eminent role in pathophysiology of infection and inflammation. Their actual clinical importance is, however, uncertain. In this study, we tested the hypothesis that inflammatory cytokines could be useful in detection of infections in high-risk patients. We prospectively studied the diagnostic value of determination of concentrations of interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), and the 55 and 75-kd soluble TNF receptors (sTNFR-p55 and sTNFR-p75) in detection of nosocomial infections in 52 patients with acute ischemic stroke, as an exemplary high-risk group, and compared these findings to those of conventional inflammatory indicators of inflammation (C-reactive protein and leukocyte count). After 1 week of hospitalization, 27% of the patients had minor or moderately severe nosocomial infections. This subpopulation exhibited significantly increased concentrations of IL-6 and sTNFR-p55 but not of IL-1beta, TNF-alpha, or sTNFR-p75. As expected, levels of C-reactive protein and leukocytes were increased in infected patients. The sensitivity and specificity for detection of nosocomial infections at day 7 of hospitalization was highest for IL-6, followed by C-reactive protein and the leukocyte count. The data suggest that the proinflammatory cytokine IL-6, in addition to its considerable pathophysiologic importance in systemic inflammation, may be valuable in detection of infections in high-risk patients. PMID- 9390643 TI - Heterozygosity for the Leiden mutation of the factor V gene, a common pathoetiology for osteonecrosis of the jaw, with thrombophilia augmented by exogenous estrogens. AB - We assessed whether heterozygosity for the thrombophilic Leiden mutation of the factor V gene (MFV) was pathogenetic for alveolar osteonecrosis of the jaw and chronic facial pain (neuralgia-inducing cavitational osteonecrosis (NICO)) in 89 patients with NICO. A second specific aim was to assess for thrombophilic synergism between exogenous estrogens and MFV for development of osteonecrosis of the jaw. MFV was found in 24% of the patients, 16 (21%) of 76 women and 5 (39%) of 13 men. The mutation was much less common in healthy normal controls: 3 (3%) of 101 women (chi2 = 14.8, p = 0.001) and 4 (3.7%) of 108 men (chi2 = 20.4, p = 0.001). Patients with and without MFV did not differ in tissue plasminogen activator activity, plasminogen activator inhibitor activity, proteins C and S, lipoprotein (a), or anticardiolipin antibodies (p > 0.05). Use of standard-dose oral contraceptives and/or postmenopausal estrogens before the development of NICO was more common in female patients with MFV (13 (81%) of 16) than in those without it (23 (38%) of 60; chi2 = 9.33, p = 0.002). When the thrombophilic effects of such exogenous estrogens were superimposed on the familial resistance to activated protein C associated with MFV, thrombophilia was augmented and the risk of osteonecrosis was increased. Since heterozygosity for this mutation occurs in at least 3% of unselected, healthy women, measurement of resistance to activated protein C and MFV would identify women at high risk for venous thrombosis and osteonecrosis, in whom use of oral contraceptives or postmenopausal estrogens might be contraindicated, while identifying a much larger group of women (approximately 97%) without the mutation whose risk from exogenous estrogens would be low. PMID- 9390644 TI - Neuroprotective effects of inhibiting poly(ADP-ribose) synthetase on focal cerebral ischemia in rats. AB - Poly(adenosine 5'-diphosphoribose) synthetase (PARS) has been described as an important candidate for mediation of neurotoxicity by nitric oxide. In the current study, we demonstrate for the first time that in vivo administration of a potent PARS inhibitor, 3,4-dihydro 5-[4-1(1-piperidinyl) butoxy]-1(2H) isoquinolinone, leads to a significant reduction of infarct volume in a focal cerebral ischemia model in the rat. Focal cerebral ischemia was produced by cauterization of the right distal middle cerebral artery (MCA) with bilateral temporary common carotid artery occlusion for 90 minutes. 3,4-Dihydro 5[4-(1 piperidinyl) butoxy]-1(2H)-isoquinolinone was dissolved in dimethyl sulfoxide and injected intraperitoneally. Animals were treated 2 hours before MCA occlusion (control, n = 14; 5 mg/kg, n = 7; 10 mg/kg, n = 7; 20 mg/kg, n = 7; 40 mg/kg, n = 7), and 2 hours after MCA occlusion (same doses as before treatment). Twenty-four hours after MCA occlusion, the total infarct volume was measured using 2,3,5 triphenyltetrazolium chloride. Inhibition of PARS leads to a significant decrease in the damaged volume in the 5 mg/kg-treated group (106.7 +/- 23.2 mm3; mean +/- SD, P < 0.002), the 10 mg/kg-treated group (76.4 +/- 16.8 mm3, P < 0.001), and the 20 mg/kg-treated group (110.2 +/- 42.0 mm3, P < 0.02) compared with the control group (165.2 +/- 34.0 mm3). The substantial reduction in infarct volume indicates that the activation of PARS may play an important role in the pathogenesis of brain damage in cerebral ischemia through intracellular energy depletion. PMID- 9390645 TI - Ischemic brain injury is mediated by the activation of poly(ADP ribose)polymerase. AB - Poly(ADP-ribose)polymerase (PARP, EC 2.4.2.30), an abundant nuclear protein activated by DNA nicks, mediates cell death in vitro by nicotinamide adenine dinucleotide (NAD) depletion after exposure to nitric oxide. The authors examined whether genetic deletion of PARP (PARP null mice) or its pharmacologic inhibition by 3-aminobenzamide (3-AB) attenuates tissue injury after transient cerebral ischemia. Twenty-two hours after reperfusion following 2 hours of filamentous middle cerebral artery occlusion, ischemic injury was decreased in PARP-/- and PARP+/- mice compared with PARP+/+ litter mates, and also was attenuated in 129/SV wild-type mice after 3-AB treatment compared with controls. Infarct sparing was accompanied by functional recovery in PARP-/- and 3-AB-treated mice. Increased poly(ADP-ribose) immunostaining observed in ischemic cell nuclei 5 minutes after reperfusion was reduced by 3-AB treatment. Levels of NAD--the substrate of PARP--were reduced 2 hours after reperfusion and were 35% of contralateral levels at 24 hours. The decreases were attenuated in PARP-/- mice and in 3-AB-treated animals. Poly(ADP-ribose)polymerase cleavage by caspase-3 (CPP-32) has been proposed as an important step in apoptotic cell death. Markers of apoptosis, such as oligonucleosomal DNA damage, total DNA fragmentation, and the density of terminal deoxynucleotidyl transferase dUTP nick-end-labelled (TUNEL +) cells, however, did not differ in ischemic brain tissue of PARP-/- mice or in 3-AB-treated animals versus controls, although there were differences in the number of TUNEL-stained cells reflecting the decrease in infarct size. Thus, ischemic brain injury activates PARP and contributes to cell death most likely by NAD depletion and energy failure, although the authors have not excluded a role for PARP in apoptotic cell death at earlier or later stages in ischemic cell death. Inhibitors of PARP activation could provide a potential therapy in acute stroke. PMID- 9390646 TI - Calcium-activated K+ channels in cerebral arterioles in piglets are resistant to ischemia. AB - Our previous studies indicate that function of ATP-dependent K+ channels (K(ATP)) in cerebral arterioles is suppressed after ischemia. In the current study, we examined pial arteriolar responses to forskolin, dibutyryl-cAMP, NS-1619, and methionine (met)-enkephalin, activators of calcium-dependent K+ channels (K(Ca)) before and 1 hour after 10 minutes of total, global ischemia in anesthetized piglets. Arteriolar diameters were measured using a closed cranial window and intravital microscopy. All pharmacologic agents were given topically. Baseline diameters were approximately 100 microm, and diameters had returned to normal by 1 hour after ischemia. Forskolin dilated arterioles by 9 +/- 3%, 18 +/- 4%, and 31 +/- 12% at 5 x 10(-8), 5 x 10(-7), and 10(-6) mol/L, respectively (P < 0.05, n = 10). In addition, dibutyryl-cAMP dilated arterioles by 8 +/- 2% at 10(-4) mol/L and 14 +/- 2% at 3 x 10(-4) mol/L (P < 0.05, n = 6). Also, NS-1619 increased diameter of arterioles by 9 +/- 2% at 10(-7) mol/L and 17 +/- 9% at 10(-5) mol/L (P < 0.05, n = 5). Finally, met-enkephalin dilated arterioles by 9 +/- 2% at 10( 8) mol/L and 16 +/- 3% at 10(-6) mol/L (P < 0.05, n = 5). At 1 hour after ischemia, arteriolar dilator effects to forskolin, dibutyryl-cAMP and NS-1619, and met-enkephalin were intact. Thus, in contrast to K(ATP), K(Ca) in cerebral arterioles are resistant to ischemic stress. PMID- 9390647 TI - Endothelin-B receptors in cerebral resistance arterioles and their functional significance after focal cerebral ischemia in cats. AB - In the cerebral circulation, endothelin-A receptor activation mediates marked prolonged vasoconstriction whereas endothelin-B (ETB) receptor activation effects dilation. In contrast to some peripheral vascular beds, ET(B) receptor-induced vasoconstriction has not yet been demonstrated in brain vessels. In this study in chloralose-anesthetized cats, with perivascular microapplications of ET(B) selective agonist (BQ-3020) and antagonist (BQ-788), we investigated whether ET(B) receptor-mediated constriction could be uncovered in cortical arterioles in vivo. In addition, we examined whether normal dilator response to ET(B) receptor activation is preserved in postischemic cerebral arterioles. The first microapplication of the selective ET(B) receptor agonist BQ-3020 (1 micromol/L) onto a pial cortical arteriole elicited marked dilation (caliber increased by 26.3 +/- 15.1% from preinjection baseline). A second application of BQ-3020 (10 minute interval) onto the same vessel failed to evoke any significant vasomotor response. Subsequent (third and fourth) adventitial microapplication of the ET(B) receptor agonist on the same arteriolar site effected a significant constriction of cerebral arterioles (-15.3 +/- 12.7% and -9.7 +/- 6.3% from preinjection baseline, respectively, at 20 and 30 minutes after the first application). The pial arterioles did not display tachyphylaxis to repeated applications of potassium (10 mmol/L). The perivascular application of the ET(B) receptor antagonist BQ-788 (0.001 to 1 micromol/L) had no effect on arteriolar caliber per se but blocked both BQ-3020-induced dilation (inhibitory concentration approximately 5 nmol/L) and vasoconstriction elicited by repeated activation of ET(B) receptors. After middle cerebral artery occlusion, most of the arterioles examined displayed a sustained dilation. The microapplication of BQ-3020 into the perivascular space surrounding postischemic dilated arterioles elicited a constriction of a similar magnitude to that induced by application of CSF (-17 +/ 7% and -17 +/- 7% from preinjection baseline, respectively). The adventitial microapplication of the ET(B) receptor antagonist (BQ-788, 0.1 micromol/L) on postocclusion dilated pial arterioles effected no change in the arteriolar caliber when compared with preinjection baseline. This BQ-788-induced response was significantly different from that induced by perivascular microinjection of CSF (P < 0.001, analysis of variance). These investigations indicate that (1) repeated activation of ET(B) receptors displays tachyphylaxis of the vasodilator response but also uncovers significant constriction of cerebral arterioles in vivo; (2) the ability of BQ-3020 to elicit dilation is lost within 30 minutes of induced focal ischemia; and (3) ET(B)-mediated contractile tone contributes in a small but significant manner in limiting postischemia dilation of cortical pial arterioles. PMID- 9390648 TI - Activity of mitochondrial respiratory chain enzymes after transient focal ischemia in the rat. AB - Previous results demonstrated that after 2-hour middle cerebral artery occlusion (MCAO) in the rat, 1- to 2-hour recirculation temporarily restored the bioenergetic state and mitochondrial function, but secondary deterioration took place after 4 hours. The authors measured the activity of mitochondrial respiratory chain complexes, citrate synthase, and glutamate dehydrogenase as possible targets of secondary damage. Focal and penumbral tissues were sampled in the control condition, after 2 hours of MCAO, and after 1, 2, or 4 hours of postischemic recirculation; two groups were treated with alpha-phenyl-N-tert butyl-nitrone (PBN). Complex IV activity transiently decreased after MCAO, but after recirculation all measured activities returned to control values. PMID- 9390649 TI - Profiles of cortical tissue depolarization in cat focal cerebral ischemia in relation to calcium ion homeostasis and nitric oxide production. AB - Cortical depolarization was investigated in a topographic gradient of ischemic density after 1-hour transient middle cerebral artery occlusion in halothane anesthetized cats. A laser Doppler flow probe, an ion-selective microelectrode, and a nitric oxide (NO) electrode measured regional CBF (rCBF), direct current (DC) potential, extracellular Ca2+ concentration ([Ca2+]o), and NO concentration in ectosylvian and suprasylvian gyri of nine animals. Recordings revealed 12 of 18 sites with persistent negative shifts of the DC potential, severe rCBF reduction, and a drop of [Ca2+]o characteristic for core regions of focal ischemia. Among these sites, two types were distinguished by further analysis. In Type 1 (n = 5), rapid, negative DC shifts resembled anoxic depolarization as described for complete global ischemia. In this type, ischemia was most severe (8.9 +/- 2.5% of control rCBF), [Ca2+]o dropped fast and deepest (0.48 +/- 0.20 mmol/L), and NO concentration increased transiently (36.1 +/- 24.0 nmol/L at 2.5 minutes), and decreased thereafter. In Type 2 (n = 7), the DC potential fell gradually over the first half of the ischemic episode, rCBF and [Ca2+]o reductions were smaller than in Type 1 (16.2 +/- 8.2%; 0.77 +/- 0.41 mmol/L), and NO increased continuously during ischemia (53.1 +/- 60.4 nmol/L at 60 minutes) suggesting that in this type NO most likely exerts its diverse actions on ischemia-threatened tissue. In the remaining six recording sites, a third type (Type 3) attributable to the ischemic periphery was characterized by minimal DC shifts, mild ischemia (37.2 +/- 13.3%), nonsignificant alterations of [Ca2+]o, but decreased NO concentrations during middle cerebral artery occlusion. Reperfusion returned the various parameters to baseline levels within 1 hour, the recovery of [Ca2+]o and NO concentration being delayed in Type 1. An NO synthase inhibitor (N(G)-nitro-L-arginine, 50 mg/kg intravenously; four animals) abolished NO elevation during ischemia. In conclusion, even in the core of focal cerebral ischemia and reperfusion, different ischemic densities produce different types of cortical tissue manifesting distinctive chronological profiles of depolarization, Ca2+ influx, and NO synthesis. PMID- 9390650 TI - The effect of nitric oxide synthase inhibition on acute platelet accumulation and hemodynamic depression in a rat model of thromboembolic stroke. AB - The relative importance of hemodynamic factors in the pathogenesis of thrombotic or embolic stroke is unclear. Of particular therapeutic interest are those substances that facilitate vasodilation and the clearance of platelet aggregates in the compromised microvasculature. A likely contributor to these functions is nitric oxide because it is known to inhibit platelet aggregability and promote vascular relaxation. To investigate the involvement of nitric oxide in the hemodynamic changes after experimental ischemia, photochemically induced nonocclusive common carotid artery thrombosis (CCAT) was studied. CCAT is a rat model of unilateral carotid artery stenosis and platelet embolization to the brain. This study characterized the acute hemodynamic consequences of CCAT and the resultant pattern of platelet deposits with and without nitric oxide synthase inhibition by nitro-L-arginine methyl ester (L-NAME). In addition, the subacute local cerebral blood flow changes were studied at 24 hours. Right CCAT was produced in 30 male Wistar rats injected with (111)In-labeled platelets. Between 5 and 15 minutes after thrombosis, rats were treated with either 15 mg/kg of L NAME (intravenously) or saline vehicle. Hemodynamic changes were studied 30 to 45 minutes after thrombosis using [14C]iodoantipyrine autoradiography. Eight coronal levels were analyzed, and cortical and subcortical regions of interest were defined. Significant increases were observed in total platelets in the ipsilateral hemisphere after L-NAME treatment, and in the distribution of platelets in the anterior frontal and occipital cortices with nitric oxide synthase inhibition, encompassing the anterior and posterior border zone areas of the ipsilateral cortex. Otherwise, foci of labeled platelets were detected throughout the ipsilateral and contralateral hemispheres. Mean local cerebral blood flow images (n = 5) revealed a moderate bilateral global reduction in flow acutely, which normalized in the untreated thrombosed group by 24 hours. In contrast, the L-NAME-treated groups (sham and experimental) had lasting, widespread reductions in flow of approximately 25%. Pairwise comparisons between groups showed that CCAT/L-NAME was significantly different from shams in the corpus callosum and different from L-NAME shams in the internal capsule (P < 0.05) These hemodynamic and platelet accumulation changes may partially account for the aggravation of cognitive and sensorimotor deficits previously reported in this model of thromboembolic stroke. PMID- 9390651 TI - Local uncoupling of the cerebrovascular and metabolic responses to somatosensory stimulation after neuronal nitric oxide synthase inhibition. AB - It has recently been shown, using either genetically engineered mutant mice (nitric oxide synthase [NOS] knockout) or specific pharmacological tools, that type I NOS (neuronal isoform of NOS, [nNOS]) participates in coupling cerebral blood flow to functional activation. However, it has not been clearly established whether the associated metabolic response was preserved under nNOS inhibition and whether this action was exerted homogeneously within the brain. To address these issues, we analyzed the combined circulatory and metabolic consequences of inhibiting the nNOS both at rest and during functional activation in the rat anesthetized with alpha-chloralose. Cerebral blood flow and cerebral glucose use (CGU) were measured autoradiographically using [14C]iodoantipyrine and [14C]2 deoxyglucose during trigeminal activation induced by unilateral whiskers stimulation in vehicle- and 7-nitroindazole-treated rats. Our data show that inhibition of nNOS globally decreased CBF without altering CGU, indicating that NO-releasing neurons play a significant role in maintaining a resting cerebrovascular tone in the whole brain. During whisker stimulation, nNOS inhibition totally abolished the cerebrovascular response only in the second order relay stations (thalamus and somatosensory cortex) of the trigeminal relay without altering the metabolic response. These findings provide evidence that the involvement of neurally-derived NO in coupling flow to somatosensory activation is region-dependent, and that under nNOS inhibition, CBF and CGU may vary independently during neuronal activation. PMID- 9390652 TI - Functional activation of cerebral blood flow after cardiac arrest in rat. AB - After a period of global cerebral ischemia, CO2 reactivity and the hemodynamic metabolic activation to functional stimulation are transiently suppressed. This raises the question of whether the impaired functional coupling reflects disturbances of functional integrity of the brain or an impaired cerebrovascular reactivity. We, therefore, compared the recovery of CO2 reactivity with that of somatosensory evoked potentials, functional flow activation and neurologic deficits in a rodent model of cardiac arrest-induced cerebral ischemia, followed by up to 7 days of reperfusion. Cardiac arrest of 10 minutes' duration was produced in 24 animals by electrical fibrillation of the heart. Five animals were sham-operated controls. Resuscitation was performed by external cardiac massage, using standard resuscitation procedures. Functional activation was carried out under chloralose anesthesia by electrical stimulation of forepaws. CO2 reactivity was tested by ventilation of animals with 6% CO2. During functional and hypercapnic stimulation CBF was measured in the somatosensory cortex using laser Doppler flowmetry, and at the end of the experiment by 14C-iodoantipyrine autoradiography. Neurologic deficits were scored by evaluating consciousness and various sensory and motor functions. In control animals 6% CO2 increased CBF measured by laser-Doppler flowmetry by 28.8% +/- 8.7%. Forepaw stimulation generated somatosensory evoked potentials with an amplitude of 750 +/- 217 microV and increased CBF measured by laser-Doppler flowmetry by 86.0% +/- 18.1%. After return of spontaneous circulation, CO2 reactivity was transiently reduced to about 30% of control at 1 hour of reperfusion (P < 0.05) but returned to near control at 5 hours. Somatosensory evoked potential amplitudes were reduced to 15% of control at 45 minutes of reperfusion and returned to only 50% to 60% at 3 and 7 days after return of spontaneous circulation (P < 0.05). Functional activation of blood flow was completely suppressed during the first hour after return of spontaneous circulation but also recovered to 50% to 60% of control at 3 days after return of spontaneous circulation (P < 0.05). Linear regression analysis revealed a significant correlation between recovery of functional activation of blood flow and both recovery of the amplitude of somatosensory evoked potentials (P = 0.03) and the neurologic deficit score (P = 0.02), but not between neurologic deficit score and recovery of CO2 reactivity or somatosensory evoked potential amplitudes. These data demonstrate that the suppression of functional activation of blood flow after 10 minutes cardiac arrest is not related to impairment of coupling mechanisms but reflects ongoing disturbances of the functional integrity of the brain. Assessment of functional flow coupling is a reliable way to study postischemic recovery of the brain. PMID- 9390653 TI - Early white blood cell dynamics after traumatic brain injury: effects on the cerebral microcirculation. AB - Increasing clinical and experimental evidence suggests that traumatic brain injury (TBI) elicits an acute inflammatory response. In the present study we investigated whether white blood cells (WBC) are activated in the cerebral microcirculation early after TBI and whether WBC accumulation affects the posttraumatic cerebrovascular response. Twenty-four anesthetized rabbits had chronic cranial windows implanted 3 weeks before experimentation. Animals were divided into four experimental groups and were studied for 7 hours (groups I, IIa, and III) or 2 hours (group IIb). Intravital fluorescence videomicroscopy was used to visualize WBC (rhodamine 6G, intravenously), pial vessel diameters, and blood-brain barrier (BBB) integrity (Na+-fluorescein) at 6 hours (groups I, IIa, and III) or 1 hour (group IIb) after TBI. Group I (n = 5) consisted of sham operated animals. Groups IIa (n = 7) and IIb (n = 5) received fluid-percussion injury at 1 hour. Group III (n = 7) received fluid-percussion injury and 1 mg/kg anti-adhesion monoclonal antibody (MoAb) "IB4" 5 minutes before injury. Venular WBC sticking, intracranial pressure (ICP), and arterial vessel diameters increased significantly for 6 hours after trauma. IB4 reduced WBC margination and prevented vasodilation. Intracranial pressure was not reduced by treatment with IB4. Blood-brain barrier damage occurred at 1 hour but not at 6 hours after TBI and was independent of WBC activation. This first report using intravital videomicroscopy to study the inflammatory response after TBI reveals upregulated interaction between WBC and cerebral endothelium that can be manipulated pharmacologically. White blood cell activation is associated with pial arteriolar vasodilation. White blood cells do not induce BBB breakdown less than 6 hours after TBI and do not contribute to posttraumatic ICP elevation. The role of WBC more than 6 hours after TBI should be investigated further. PMID- 9390654 TI - Heparin inhibits leukocyte rolling in pial vessels and attenuates inflammatory changes in a rat model of experimental bacterial meningitis. AB - Heparin is a natural proteoglycan that was first described in 1916. In addition to its well characterized effect on blood coagulation, it is becoming clear that heparin also modulates inflammatory processes on several levels, including the interference with leukocyte-endothelium interaction. Anecdotal observations suggest a better clinical outcome of heparin-treated patients with bacterial meningitis. The authors demonstrate that heparin, a glycosaminoglycan, inhibits significantly in the early phase of experimental pneumococcal meningitis the increase of 1) regional cerebral blood flow (125 +/- 18 versus 247 +/- 42%), 2) intracranial pressure (4.5 +/- 2.0 versus 12.1 +/- 2.2 mm Hg), 3) brain edema (brain water content: 78.23 +/- 0.33 versus 79.49 +/- 0.46%), and 4) influx of leukocytes (571 +/- 397 versus 2400 +/- 875 cells/microL) to the cerebrospinal fluid compared with untreated rats. To elucidate the possible mechanism of this observation, the authors investigated for the first time leukocyte rolling in an inflammatory model in brain venules by confocal laser scanning microscopy in vivo. Heparin significantly attenuates leukocyte rolling at 2, 3, and 4 hours (2.8 +/- 1.3 versus 7.9 +/- 3.2/100 microm/min), as well as leukocyte sticking at 4 hours (2.1 +/- 0.4 versus 3.5 +/- 1.0/100 microm/min) after meningitis induction compared with untreated animals. The authors conclude that heparin can modulate acute central nervous system inflammation and, in particular, leukocyte endothelium interaction, a key process in the cascade of injury in bacterial meningitis. They propose to evaluate further the potential of heparin in central nervous system inflammation in basic and clinical studies. PMID- 9390655 TI - Trafficking of amino acids between neurons and glia in vivo. Effects of inhibition of glial metabolism by fluoroacetate. AB - Glial-neuronal interchange of amino acids was studied by 13C nuclear magnetic resonance spectroscopy of brain extracts from fluoroacetate-treated mice that received [1,2-(13)C]acetate and [1-(13)C]glucose simultaneously. [13C]Acetate was found to be a specific marker for glial metabolism even with the large doses necessary for nuclear magnetic resonance spectroscopy. Fluoroacetate, 100 mg/kg, blocked the glial, but not the neuronal tricarboxylic acid cycles as seen from the 13C labeling of glutamine, glutamate, and gamma-aminobutyric acid. Glutamine, but not citrate, was the only glial metabolite that could account for the transfer of 13C from glia to neurons. Massive glial uptake of transmitter glutamate was indicated by the labeling of glutamine from [1-(13)C]glucose in fluoroacetate-treated mice. The C-3/C-4 enrichment ratio, which indicates the degree of cycling of label, was higher in glutamine than in glutamate in the presence of fluoroacetate, suggesting that transmitter glutamate (which was converted to glutamine after release) is associated with a tricarboxylic acid cycle that turns more rapidly than the overall cerebral tricarboxylic acid cycle. PMID- 9390656 TI - Imaging experimental brain tumors with 1-aminocyclopentane carboxylic acid and alpha-aminoisobutyric acid: comparison to fluorodeoxyglucose and diethylenetriaminepentaacetic acid in morphologically defined tumor regions. AB - The goal of this study was to evaluate the differences and define the advantages of imaging experimental brain tumors in rats with two nonmetabolized amino acids, 1-aminocyclopentane carboxylic (ACPC) acid and alpha-aminoisobutyric (AIB) acid compared with imaging with fluorodeoxyglucose (FDG) or the gallium diethylenetriaminepentaacetic acid chelate (Ga-DTPA). 1-aminocyclopentane carboxylic acid, AIB, and FDG autoradiograms were obtained 60 minutes after intravenous injection to simulate positron emission tomography (PET) imaging, whereas the Ga-DTPA autoradiograms were obtained 5 or 10 minutes after injection to simulate gadolinium (Gd)-DTPA-enhanced magnetic resonance (MR) images. Three experimental tumors were studied (C6, RG2, and Walker 256) to provide a range of tumor types. Triple-label quantitative autoradiography was performed, and parametric images of the apparent distribution volume (Va, mL/g) for ACPC or AIB, relative glucose metabolism (R, micromol/100 g/min), vascular permeability to Ga DTPA (K1, microL/min/g), and histology were obtained from the same tissue section. The four images were registered in an image array processor, and regions of interest in tumor and contralateral brain were defined on morphologic criteria (histology) and were transferred to the autoradiographic images. A comparative analysis of all measured values was performed. The location and morphologic characteristics of the tumor had an effect on the images and measurements of Va, R, and K1. Meningeal extensions of all three tumors consistently had the highest amino acid uptake (Va) and vascular permeability (K1) values, and subcortical portions of the tumors usually had the lowest values. Va and R (FDG) values generally were higher in tumor regions with high-cell density and lower in regions with low-cell density. Tumor areas identified as "impending" necrosis on morphologic criteria consistently had high R values, but little or no change in Va or K1. Tumor necrosis was seen consistently only in the larger Walker 256 tumors; low values of R and Va for AIB (less for ACPC) were measured in the necrotic-appearing regions, whereas K1 was not different from the mean tumor value. The highest correlations were observed between vascular permeability (K1 for Ga-DTPA) and Va for AIB in all three tumors; little or no correlation between vascular permeability and R was observed. The advantages of ACPC and AIB imaging were most convincingly demonstrated in C6 gliomas and in Walker 256 tumors. 1 aminocyclopentane was substantially better than FDG or Ga-DTPA for identifying tumor infiltration of adjacent brain tissue beyond the macroscopic border of the tumor; ACPC also may be useful for identifying low-grade tumors with an intact blood-brain barrier. Contrast-enhancing regions of the tumors were visualized more clearly with AIB than with FDG or Ga-DTPA; viable and necrotic-appearing tumor regions could be distinguished more readily with AIB than with FDG. [11C] labeled ACPC and AIB are likely to have similar advantages for imaging human brain tumors with PET. PMID- 9390657 TI - In vivo regulation of DOPA decarboxylase by dopamine receptors in rat brain. AB - To test the hypothesis that dopamine (DA) receptors influence cerebral DOPA decarboxylase (DDC) activity in vivo, we used HPLC to measure the kinetics of the cerebral uptake and metabolism of [3H]DOPA in carbidopa-treated rats, and in rats also treated acutely with a DA receptor antagonist (flupenthixol, 2 mg/kg, intraperitoneally) or a DA receptor agonist (apomorphine, 200 microg/g, subcutaneously). The unidirectional blood-brain clearance of [3H]DOPA (K1DOPA, 0.030 mL g(-1) min(-1)) increased by 50% after flupenthixol. The magnitudes of the relative DDC activity (k3DOPA) in striatum (0.20 min(-1)), olfactory tubercle (0.11 min(-1)), and hypothalamus (0.15 min(-1)) of carbidopa-treated rats were doubled with flupenthixol, but cortical DDC activity was unaffected (0.02 min( 1)). Apomorphine reduced the magnitude of k3DOPA in striatum by 20%. The rate constant for catabolism of [3H]DA formed in brain (k7', monoamine oxidase [MAO] activity), which ranged from 0.025 min(-1) in striatum to 0.08 min(-1) in hypothalamus of carbidopa-treated rats, globally increased 2- to 4-fold after flupenthixol, and decreased to 0.003 min(-1) in striatum after apomorphine. These in vivo results confirm the claim that acute blockade of DA receptors with flupenthixol stimulates the synthesis of [3H]DA from [3H]DOPA, and that this [3H]DA is subject to accelerated catabolism. Conversely, activation of the DA receptors with apomorphine inhibits DDC activity and DA catabolism. PMID- 9390658 TI - Effects of extracranial radioactivity on measurement of cerebral glucose metabolism by rat-PET with [18F]-2-fluoro-2-deoxy-D-glucose. PMID- 9390659 TI - AAEM minimonograph #26: the electrodiagnosis of carpal tunnel syndrome. American Association of Electrodiagnostic Medicine. AB - The electrodiagnosis of carpal tunnel syndrome (CTS) is reviewed, including discussions of old and new techniques of motor and sensory nerve conduction, anomalous innervation, and needle electrode examination. A variety of sensitive nerve conduction studies (NCSs) are available for the evaluation of a patient with suspected CTS. For any particular patient, the NCS method chosen by the clinical neurophysiologist may vary for a number of reasons, including the severity of the deficit and the presence of superimposed conditions. PMID- 9390660 TI - Interaction of thyroid state and denervation on skeletal myosin heavy chain expression. AB - The goal of this study was to examine the effects of altered thyroid state and denervation (Den) on skeletal myosin heavy chain (MHC) expression in the plantaris and soleus muscles. Rats were subjected to unilateral denervation (Den) and randomly assigned to one of three groups: (1) euthyroid; (2) hyperthyroid; (3) and hypothyroid. Denervation caused severe muscle atrophy and muscle-type specific MHC transformation. Denervation transformed the soleus to a faster muscle, and its effects required the presence of circulating thyroid hormone. In contrast, denervation transformed the plantaris to a slower muscle independently of thyroid state. Furthermore, thyroid hormone effects did not depend upon innervation status in the soleus, while they required the presence of the nerve in the plantaris. Collectively, these findings suggest that both thyroid hormone and intact nerve (a) differentially affect MHC transformations in fast and slow muscle; and (b) are important factors in regulating the optimal expression of both type I and IIB MHC genes. This research suggests that for patients with nerve damage and/or paralysis, both muscle mass and biochemical properties can also be affected by the thyroid state. PMID- 9390661 TI - Smooth muscle electromyography from rat urethra. AB - Electrical signals recorded from the penis have been suggested as reflecting electromyographic activity in the underlying smooth muscles. In order to verify this assertion, we manipulated the signal recorded from rat urethra surface. Stimulation of the pelvic nerve brought about a reduction of activity (965 +/- 826 to 166 +/- 143 microV, root mean square of the power at range 0.005-1 Hz, P = 0.008), with a significant frequency-response relationship (P = 0.0002). This effect was not altered by temporary closure of the aorta (P = 0.89), thus ruling out hemodynamic artifact as a possible cause of signal change during stimulation. Our findings support the assertion that the signal indeed reflects activity in smooth muscle. PMID- 9390662 TI - A randomized, controlled trial of creatine monohydrate in patients with mitochondrial cytopathies. AB - Fatigue in patients with mitochondrial cytopathies is associated with decreased basal and postactivity muscle phosphocreatine (PCr). Creatine monohydrate supplementation has been shown to increase muscle PCr and high-intensity power output in healthy subjects. We studied the effects of creatine monohydrate administration (5 g PO b.i.d. x 14 days --> 2 g PO b.i.d. x 7 days) in 7 mitochondrial cytopathy patients using a randomized, crossover design. Measurements included: activities of daily living (visual analog scale); ischemic isometric handgrip strength (1 min); basal and postischemic exercise lactate; evoked and voluntary contraction strength of the dorsiflexors; nonischemic, isometric, dorsiflexion torque (NIDFT, 2 min); and aerobic cycle ergometry with pre- and post-lactate measurements. Creatine treatment resulted in significantly (P < 0.05) increased handgrip strength, NIDFT, and postexercise lactate, with no changes in the other measured variables. We concluded that creatine monohydrate increased the strength of high-intensity anaerobic and aerobic type activities in patients with mitochondrial cytopathies but had no apparent effects upon lower intensity aerobic activities. PMID- 9390663 TI - Computing normative ranges without recruiting normal subjects. AB - Computing normative data by recruiting normal subjects is extremely difficult. However, many who are examined in a typical clinical neurophysiology lab are normal. In this study we show how to use this abundance of referred subjects to compute normative distal latency statistics from the values themselves. If all latencies are displayed on a frequency distribution, the very left side, the shorter latencies, belong to the left side of the Gaussian "bell" of normal subjects. By curve-fitting that side one can compute the coefficients of the latter. We started with an initial range of 2.0-3.6 ms and then recursively added data points until a "goodness-of-fit" criterion maximized. We computed these coefficients from 982 median motor distal latencies, showing highly significant fit (P < 0.00001): mean at 3.76 ms and SD of 0.45 ms. The data analyzed in this study are only an example of the technique. The results are unique in that they were derived mathematically, without using normal subjects. PMID- 9390664 TI - Abscence of laminin alpha1 chain in the skeletal muscle of dystrophic dy/dy mice. AB - In Duchenne muscular dystrophy (DMD) and laminin alpha2 defective congenital muscular dystrophies (CMD) there are reports of an induction of laminin alpha1 chain in regenerating muscle fibers. These studies are based on immunohistochemistry data with one monoclonal antibody alone. Based on these data we sought to establish if the laminin alpha1 chain is induced in the muscle of dy/dy mice. We found no evidence of induction of laminin alpha1 chain protein or mRNA in dystrophic dy/dy skeletal muscle fibers as determined by immunohistochemistry, Western blotting, Northern blotting, or PCR analysis. Our data point to the need for additional immunological reagents specific for human laminin-alpha1 to resolve whether the conflicting data on laminin-alpha1 distribution in human and mouse tissues is due to species differences or, alternatively, due to differences in reagent specificity. Our data might be important when designing therapy strategies for CMD. PMID- 9390665 TI - Concentric and single fiber electrode spatial recording characteristics. AB - A better appreciation of the specific spatial recording characteristics of the single fiber and concentric needle electrode can result in more accurate physiologic and theoretical interpretations of single fiber and quantitative motor unit action potential analysis. We demonstrate by physical modeling that the 90% and 99% amplitude sensitivity envelopes are not simple hemispherical shapes. The 90% sensitivity concentric electrode volume does not extend beyond the insulated portion of the 15 degree beveled surface between the core and cannula and extends only 280 microm perpendicularly from the center of the core's surface. The 99% envelope extends approximately 830 microm perpendicularly from the core's center. This is a much smaller volume of sensitivity than exists for a similarly modeled monopolar electrode. The 90% and 99% envelopes extend to 110 and 320 microm perpendicularly from the exposed single fiber core. Both the single fiber and concentric needle volumes of sensitivity have specific asymmetries described. PMID- 9390666 TI - Human leukocyte antigen class I in polymyositis: leukocyte infiltrates, regeneration, and impulse block. AB - In polymyositis (PM), T-cell mediated myocytotoxicity is directed against strongly human leukocyte antigen class I positive (HLA-I+) muscle fibers. Fiber regeneration probably is partly responsible for this HLA-I up-regulation. We have evaluated regeneration, denervation/impulse blockade, and focal leukocyte infiltrates as possible HLA-I inducing factors in PM. Distinctive patterns of HLA I, nerve cell adhesion molecule (NCAM), and vimentin expression accompany denervation and regeneration. Regenerating fibers also have centralized nuclei. Using semiquantitative methods, we examined strongly HLA-I+ fibers in PM muscle biopsies for these markers. Sarcoplasmic HLA-I levels were related to the presence of leukocyte infiltrates and invasion of fibers. Strongly HLA-I+ fibers were frequently invaded, and regeneration-associated changes were usually observed at sites of fiber damage. Sarcoplasmic HLA-I levels were stable along intact fibers, also adjacent to leukocyte infiltrates. A majority of the strongly HLA-I+ fibers were nonregenerating (NCAM+ only). Though other mechanisms cannot be excluded, this suggests that impulse blockade or denervation may contribute to extra HLA-I up-regulation in these fibers. PMID- 9390667 TI - Acetazolamide reduces peripheral afferent transmission in humans. AB - Carbonic anhydrase has been localized in skeletal muscle and nerve, thus, inhibition with acetazolamide (ACZ) may alter nerve and/or muscle function in healthy humans. ACZ (3 oral doses 14, 8, and 2 h prior to testing) reduced isometric force (37%) and peak to peak electromyographic (EMG) amplitude (1.38 mV to 0.83 mV), while increasing EMG latency associated with a unilateral Achilles tendon-tap. Reflex recovery profiles, following a contralateral conditioning tap, were similar in both placebo and ACZ experiments. ACZ led to significant changes in Hmax/Mmax ratio (52.19/14.42 to 45.73/15.65) and H-reflex latency (34.18 +/- 2.54 ms to 35.24 +/- 2.74 ms). Motor nerve conduction velocity and maximal voluntary isometric torque (knee extensors) were unaltered by ACZ. These data suggest that inhibition of the tendon-tap reflex and associated isometric force, following ACZ, is related to impairment of synaptic integrity between la fibers of the muscle spindle and the alpha motor neuron and not impairment of the muscle spindle or force-generating capacity. PMID- 9390668 TI - Afferent-inherent regulation of myosin heavy chain isoforms in rat muscle spindles. AB - Whether afferents exert their morphogenetic influence on spindles through release of trophic factors at intrafusal fiber junctions or via participation in proprioceptive pathways which modulate the motor activity to muscles was investigated by comparing myosin heavy chain (MHC) expression in intrafusal fibers after ablation of afferents (deafferentation, or DA) to the extensor digitorum longus (EDL) of adult rats or after ablation of the corresponding central processes of afferents to the spinal cord (central-process ablation, or CPA). DA and CPA elicited an exaggerated pedal plantarflexion, and hypertrophy of the EDL concomitant with atrophy of the soleus in the affected hindlimb. Frequencies and patterns of expression of seven MHCs expressed by intrafusal fibers in CPA muscles were indistinguishable from normal rats. However, frequencies and patterns of expression of several MHCs were abnormal following DA. Thus factors transported anterogradely from afferents to intrafusal fibers may regulate MHC expression in intrafusal fibers. PMID- 9390669 TI - Effect of age and gender on sudomotor and cardiovagal function and blood pressure response to tilt in normal subjects. AB - Normative data are limited on autonomic function tests, especially beyond age 60 years. We therefore evaluated these tests in a total of 557 normal subjects evenly distributed by age and gender from 10 to 83 years. Heart rate (HR) response to deep breathing fell with increasing age. Valsalva ratio varied with both age and gender. QSART (quantitative sudomotor axon-reflex test) volume was consistently greater in men (approximately double) and progressively declined with age for all three lower extremity sites but not the forearm site. Orthostatic blood pressure reduction was greater with increasing age. HR at rest was significantly higher in women, and the increment with head-up tilt fell with increasing age. For no tests did we find a regression to zero, and some tests seem to level off with increasing age, indicating that diagnosis of autonomic failure was possible to over 80 years of age. PMID- 9390670 TI - Chronic inflammatory demyelinating polyradiculoneuropathy in children: II. Long term follow-up, with comparison to adults. AB - We previously reviewed the presentation, initial clinical course, and electrodiagnostic features of children with chronic inflammatory demyelinating polyradiculoneuropathy (CIDP). We now report the long-term follow-up of 12 children with idiopathic CIDP, and compare these to 62 adults with idiopathic CIDP. Children often had more rapidly fluctuating courses than adults. A relapsing course was significantly more common in children than in adults. The recovery of children from each episode of deterioration was usually excellent, and better, on average, than in adults. Ventilatory support was never required for children with slowly evolving illness; only 2 children with a precipitous onset clinically resembling Guillain-Barre syndrome required ventilatory support. Prednisone, plasma exchange, and intravenous immunoglobulin (IVIg) usually were effective in children. Multiple courses of IVIg could be given with continued efficacy. Treatment often could be discontinued in children with relapsing courses. The prognosis for children was excellent. Adults demonstrated a good, but more variable, outcome. PMID- 9390671 TI - Anti-Hu-associated paraneoplastic sensory neuropathy responding to early aggressive immunotherapy: report of two cases and review of literature. AB - Anti-Hu-associated paraneoplastic sensory neuropathy (PSN) has been reported to be nonresponsive to immunotherapy or cancer therapy. We report 2 patients with anti-Hu-associated PSN who achieved sustained clinical improvement with early and aggressive immunotherapy 10-15 months before the diagnosis of small-cell lung carcinoma. Both had chronic "sensory neuronopathy plus"; in addition to sensory neuronopathy, case 1 had a motor-autonomic dysfunction with encephalopathy, and case 2 had a motor-autonomic dysfunction with swallowing difficulty. These two cases were unusual in that sustained clinical improvement was achieved with early aggressive immunotherapy before the detection of cancer and without any concomitant anticancer therapy or lowering of anti-Hu antibody titer. We believe that early and aggressive immunotherapy should be tried in any patient with anti Hu-associated PSN, as it may induce sustained clinical improvement. PMID- 9390672 TI - Remission of myasthenia gravis caused by proteinuria in nephrotic syndrome. AB - Myasthenia gravis is caused by antibodies against acetylcholine receptors and is treated with inhibition or elimination of antibody production. We report a 43 year-old myasthenic female who was symptomatic until she developed proteinuria from nephrotic syndrome, which caused a marked drop in acetylcholine receptor antibody titer with remission of myasthenia. Treatment of the nephrotic syndrome produced exacerbation of her myasthenia and a rise in antibody level. This patient's improvement is the result of antibody elimination during proteinuria in nephrotic syndrome. PMID- 9390673 TI - Reversible paralysis with status asthmaticus, steroids, and pancuronium: clinical electrophysiological correlates. AB - Prolonged neuromuscular weakness has been identified after neuromuscular blockade in intensive care unit patients on mechanical ventilation. Previously reported electromyographic studies in these patients documented both neurogenic features and features consistent with a myopathy. We recorded sequential electrophysiological parameters during recovery from neuromuscular blockade in 5 patients with clinical weakness. An evolving pattern was identified. The early features were in keeping with previous reports of neurogenic changes, and this evolved into features consistent with a primary myopathy. Several potential underlying mechanisms are discussed. PMID- 9390674 TI - Anterior interosseous nerve syndrome presenting with pronator teres weakness: a case report. AB - Anterior interosseous nerve syndrome (AINS) has been well described. A key muscle to examine clinically and on electromyography is the pronator teres, as this can differentiate between forearm and more proximal entrapment sites. We present a case of AINS with marked weakness and denervation of pronator teres. At operation the anterior interosseous nerve gave rise to the nerve to pronator teres and was entrapped by a fibrous band from the deep head of pronator teres. PMID- 9390675 TI - Fulminant demyelinating neuropathy mimicking cerebral death. AB - Guillain-Barre syndrome can very rarely present with acute quadripares and cranial nerve involvement resembling a locked-in state. We describe a very unusual case of fulminant neuropathy in a child who was previously exposed to vincristine. The clinical picture resembled brain death; however, electrodiagnostic studies led to the diagnosis of a peripheral neuropathy. Serial electrodiagnostic studies and pathologic findings confirmed demyelination. PMID- 9390676 TI - Focal myopathy associated with chronic intramuscular injection of piritramide. AB - We report a patient in whom chronic intramuscular piritramide led to a focal fibrotic myopathy. Since piritramide myotoxicity has never been reported, we have studied its effect on rat skeletal muscle. Chronic intramuscular piritramide led to fibrous connective tissue replacement of rat skeletal muscle, similar to that found in the patient's muscle. Although the pathogenetic mechanism of piritramide myopathy is unclear, we caution against prolonged intramuscular use of piritramide to prevent this potentially debilitating adverse effect. PMID- 9390677 TI - Acoustic myography. PMID- 9390679 TI - Numb chin syndrome heralding myeloma relapse. PMID- 9390678 TI - Peripheral myoclonus due to spinal root lesion. PMID- 9390680 TI - Selective inhibition of ipsilateral and contralateral R3 of the blink reflex by capsaicin. PMID- 9390681 TI - Variation of calculated ulnar motor conduction velocity across the elbow with body mass index. PMID- 9390682 TI - T cell turnover in HIV-1 disease. PMID- 9390683 TI - Exit of the pre-TCR from the ER/cis-Golgi is necessary for signaling differentiation, proliferation, and allelic exclusion in immature thymocytes. AB - A major issue is whether surface expression of the pre-TCR is necessary for signaling the development of immature thymocytes. To address this question, we generated transgenic mice expressing a TCRbeta chain that had a strong endoplasmic reticulum (ER) retrieval signal (TCRbetaER) and that was expressed intracellularly but failed to reach the cell surface. In TCRbetaER transgenic mice, there was a failure of allelic exclusion. Also, the transgene failed to rescue the developmental defects observed in TCRbeta-null mice. In contrast, TCRbeta transgenes with a mutant ER retrieval sequence or lacking this sequence signaled efficient allelic exclusion and suppressed the TCRbeta-/- defect. These data show that exit of the pre-TCR from the ER/cis-Golgi is required for progression through the double-negative thymocyte checkpoint. PMID- 9390684 TI - Essential role of the pre-T cell receptor in allelic exclusion of the T cell receptor beta locus. AB - Following the recent realization that TCR beta transgenes can severely inhibit the rearrangement of endogenous Vbeta gene segments in the absence of pre-TCR alpha (pT alpha) chains, we tested whether the pre-TCR has an essential role in TCR beta allelic exclusion under more physiological conditions by analyzing TCR rearrangement in immature thymocytes by single-cell PCR. Our results in pT alpha+ mice are consistent with an ordered model of TCR beta rearrangement beginning on one allele and continuing on the other only when the first attempt is unsuccessful. By contrast, a higher proportion of thymocytes from pT alpha-/- mice exhibited two productive TCR beta alleles. Thus, the pre-TCR-independent suppression of rearrangement by TCR beta transgenes represents a transgene artifact, whereas under physiological conditions the pre-TCR is essential for allelic exclusion. PMID- 9390686 TI - Activation of the Lck tyrosine kinase targets cell surface T cell antigen receptors for lysosomal degradation. AB - The mechanism by which TCR expression is regulated was explored by expressing a constitutively active form of the tyrosine kinase Lck (Lck505F) in T cells. Expression of Lck505F down-regulated TCR levels, an effect that was even more pronounced in CD45- T cells, in which the activity of this tyrosine kinase is further enhanced. Cells expressing Lck505F synthesized all TCR subunits, but lysosomal degradation of assembled receptors was enhanced. TCRs were rapidly internalized and degraded after removal of a tyrosine kinase inhibitor that had permitted cell surface expression. Finally, TCR levels on thymocytes were increased by an Lck inhibitor, and activation- but not phorbol ester-induced internalization of TCRs on Jurkat cells was prevented by inhibition or loss of Lck. These studies identify a regulated nonreceptor tyrosine kinase-mediated pathway for targeting cell surface receptors for lysosomal degradation. PMID- 9390685 TI - The influence of the MAPK pathway on T cell lineage commitment. AB - During development, progenitor thymocytes differentiate into either CD4 or CD8 T cells, and this fate decision depends on the specificity of the T cell antigen receptor (TCR) for MHC class II or class I molecules. Based on the mechanisms of fate specification known for simple metazoan organisms, we sought to determine whether the extracellular signal-related kinases (ERKs) play a role in T cell differentiation and lineage commitment. Using a dominant gain-of-function mutant of the erk2 gene, we show that differentiation into the CD4 lineage is favored. We also show that, conversely, the addition of a pharmacological inhibitor of the ERK pathway favors differentiation into the CD8 lineage. We present a quantitative selection model that incorporates these results as well as those of recent reports on the role of Notch in T cell lineage specification. PMID- 9390687 TI - A novel Bcl-x isoform connected to the T cell receptor regulates apoptosis in T cells. AB - We define a novel Bcl-x isoform, Bcl-x gamma, that is generated by alternative splicing and characterized by a unique 47 amino acid C-terminus. Bcl-x gamma is expressed primarily in thymocytes, where it may depend on an interaction between the TCR and host MHC products, and in mature T cells, where its expression is associated with ligation of the T cell receptor. Overexpression of Bcl-x gamma in T cells inhibits activation-induced apoptosis; inhibition of Bcl-x gamma, after stable expression of Bcl-x gamma antisense cDNA, enhances activation-induced apoptosis. In contrast to other Bcl-x isoforms, cells that fail to express Bcl-x gamma after CD3 ligation undergo programmed cell death, while activated T cells that express Bcl-x gamma are spared. Identification of Bcl-x gamma helps provide a molecular explanation of T cell activation and death after antigen engagement. PMID- 9390688 TI - A new MHC locus that influences class I peptide presentation. AB - We have investigated the HLA-B27-restricted CTL response to HY minor histocompatibility antigens in rats and mice transgenic for HLA-B27 and human beta2-microglobulin. A polymorphism was found at a locus within the H2 complex, producing two distinct but overlapping sets of B27-presented HY peptides. The locus, named Cim2, mapped between the K and Pb loci, and its product is therefore distinct from TAP, LMP, and tapasin. Identical findings in rats and mice, including identical HY peptide sequences and the failure of a rat Tap2A transgene to alter CTL recognition, suggest that a homologous locus with similar polymorphism exists in the rat. Cim2, or a closely linked locus, was found to exert a broad effect on peptide loading of both HLA-B27 and mouse class I alleles. The data thus establish a strong, previously unrecognized MHC-encoded influence on the class I antigen pathway. PMID- 9390689 TI - Growth retardation and leaky SCID phenotype of Ku70-deficient mice. AB - Ku70, Ku80, and DNA-PKcs are subunits of the DNA-dependent protein kinase (DNA PK), an enzyme implicated in DNA double-stranded break repair and V(D)J recombination. Our Ku70-deficient mice were about 50% the size of control littermates, and their fibroblasts were ionizing radiation sensitive and displayed premature senescence associated with the accumulation of nondividing cells. Ku70-deficient mice lacked mature B cells or serum immunoglobulin but, unexpectedly, reproducibly developed small populations of thymic and peripheral alpha/beta T lineage cells and had a significant incidence of thymic lymphomas. In association with B and T cell developmental defects, Ku70-deficient cells were severely impaired for joining of V(D)J coding and recombination signal sequences. These unanticipated features of the Ku70-deficient phenotype with respect to lymphocyte development and V(D)J recombination may reflect differential functions of the three DNA-PK components. PMID- 9390690 TI - Reversal of EBV immortalization precedes apoptosis in IL-6-induced human B cell terminal differentiation. AB - Cell death in B cell terminal differentiation rapidly follows cell cycle arrest in IL-6 differentiation of EBV-immortalized, IgG-bearing human lymphoblastoid cells in vitro. G1 arrest is now found to coincide with repression of EBNA2 and LMP1, two EBV genes essential for B cell transformation, without activation of the viral lytic cycle. IL-6-differentiated B cells die by apoptosis, as evidenced by increases in Annexin V binding activity, PARP cleavage, and chromatin disorganization. Expression of Mcl-1, a Bcl-2 family member, was specifically induced during IL-6 differentiation and down-regulated during apoptosis. Thus, IL 6 reverses EBV immortalization and activates the terminal differentiation program in IgG-bearing human B lymphoblastoid cells, including regulation of an anti apoptotic gene to coordinate differentiation, cell cycle arrest, and cell death. PMID- 9390691 TI - Phosphatidylinositol 3-kinase couples the interleukin-2 receptor to the cell cycle regulator E2F. AB - Cell cycle progression initiated by interleukin-2 (IL-2) in T cells is critical for lymphoproliferation and an immune response. Phosphatidyl inositol 3-kinase (PI3K) is activated by IL-2. However, nuclear targets for PI3K are not known. Here we identify the cell cycle regulator E2F as an IL-2 target in T lymphocytes and PI3K as the critical signaling pathway. We eliminate both Stat5 and Raf/MEK pathways from E2F regulation. Protein kinase B (PKB) is activated by IL-2 via PI3K. The expression of an active PKB is sufficient to induce E2F activity. Inhibition of PI3K inhibits phosphorylation of Rb, induction of cyclin D3, and degradation of p27kip1. These results establish a crucial PI3K/PKB-mediated link between the IL-2 teceptor and the cell cycle machinery. PMID- 9390692 TI - An indirect effect of Stat5a in IL-2-induced proliferation: a critical role for Stat5a in IL-2-mediated IL-2 receptor alpha chain induction. AB - Stat5a was identified as a prolactin-induced transcription factor but also is activated by other cytokines, including interleukin-2 (IL-2) and IL-7. We have now analyzed the immune system of Stat5a-deficient mice. Stat5a-/- splenocytes exhibited defective IL-2-induced expression of the IL-2 receptor alpha chain (IL 2R alpha), a protein that together with IL-2R beta and the common cytokine receptor gamma chain (gamma(c)) mediates high-affinity IL-2 binding. Correspondingly, Stat5a-/- splenocytes exhibited markedly decreased proliferation to IL-2, although maximal proliferation was still achieved at IL-2 concentrations high enough to titrate intermediate-affinity IL-2R beta/gamma(c) receptors. Thus, defective Stat5a expression results in diminished proliferation by an indirect mechanism, resulting from defective receptor expression. Correspondingly, we show that Stat5a is essential for maximal responsiveness to antigenic stimuli in vivo, underscoring the physiological importance of IL-2-induced IL-2R alpha expression. PMID- 9390693 TI - TRAF2 is essential for JNK but not NF-kappaB activation and regulates lymphocyte proliferation and survival. AB - TRAF2 is believed to mediate the activation of NF-kappaB and JNK induced by the tumor necrosis factor receptor (TNFR) superfamily, which elicits pleiotropic responses in lymphocytes. We have investigated the physiological roles of TRAF2 in these processes by expressing a lymphocyte-specific dominant negative form of TRAF2, thereby blocking this protein's effector function. We find that the TNFR superfamily signals require TRAF2 for activation of JNK but not NF-kappaB. In addition, we show that TRAF2 induces NF-kappaB-independent antiapoptotic pathways during TNF-induced apoptosis. Inhibition of TRAF2 leads to splenomegaly, lymphadenopathy, and an increased number of B cells. These findings indicate that TRAF2 is involved in the regulation of lymphocyte function and growth in vivo. PMID- 9390694 TI - Early lethality, functional NF-kappaB activation, and increased sensitivity to TNF-induced cell death in TRAF2-deficient mice. AB - TRAF2 is an intracellular signal-transducing protein recruited to the TNFR1 and TNFR2 receptors following TNF stimulation. To investigate the physiological role of TRAF2, we generated TRAF2-deficient mice. traf2-/- mice appeared normal at birth but became progressively runted and died prematurely. Atrophy of the thymus and spleen and depletion of B cell precursors also were observed. Thymocytes and other hematopoietic progenitors were highly sensitive to TNF-induced cell death and serum TNF levels were elevated in these TRAF2-deficient animals. Examination of traf2-/- cells revealed a severe reduction in TNF-mediated JNK/SAPK activation but a mild effect on NF-kappaB activation. These results suggest that TRAF2 independent pathways of NF-kappaB activation exist and that TRAF2 is required for an NF-kappaB-independent signal that protects against TNF-induced apoptosis. PMID- 9390695 TI - Lennox-Gastaut syndrome. AB - Lennox-Gastaut syndrome (LGS) is one of the intractable epilepsies of childhood that is associated with an epileptic encephalopathy. Although LGS has been accepted as a distinct epilepsy syndrome for the last 30 years, understanding of its pathogenesis is still incomplete. Because this heterogenous entity has many diverse etiologies, some with specific therapy, a complete evaluation is necessary. The natural history is well defined; most children with LGS will ultimately be mentally retarded, will continue to have seizures, and as adults will be dependent for their daily care. Therefore, their only hope is new therapies and advances in our understanding of the pathogenesis of LGS. Several new treatment options have emerged. For the first time in the last 20 years, we have several medications with documented efficacy. In addition, there are effective nonpharmacologic treatments. These treatments offer the potential for improved seizure control, which we hope will have impact and lessen the subsequent epileptic encephalopathy. Children with LGS require multidisciplinary assessment and treatment along with vigorous intervention aimed at minimizing their seizures to maximize their potential. Pediatric neurologists should be familiar with the treatments with proven efficacy, including new antiepileptic drugs, and should develop a rational plan of treatment for each child with LGS. PMID- 9390696 TI - Intraventricular urokinase for the treatment of posthemorrhagic hydrocephalus. AB - This case series pilot study assessed the safety of intraventricular urokinase administration, alternating with cerebrospinal fluid (CSF) drainage. A secondary objective was to comment on whether this therapy achieves fibrinolysis, and whether this fibrinolysis is sufficient to prevent progression of hydrocephalus to requirement for ventriculoperitoneal shunt. Six preterm infants with progressive posthemorrhagic hydrocephalus requiring treatment with a ventricular drain received an infusion of intraventricular urokinase alternating with CSF drainage for 3 days. Of the 6 treated patients, the median gestation at birth was 26.5 weeks and the median age at treatment was 30 days. One patient had an elevation in CSF erythrocyte count most likely due to successful clot lysis. One patient had an elevated CSF leukocyte count consistent with transient meningeal irritation. No other side effects were noted. Fibrinolysis was achieved in the CSF, as documented by markedly elevated D-dimer levels. Clot size diminished ultrasonographically. However, all 6 patients eventually required a ventriculoperitoneal shunt. We conclude that intermittent infusion of intraventricular urokinase alternating with periods of CSF drainage is probably a safe way to achieve a fibrinolytic state. However, when administered at the relatively late point in the neonatal course when a ventricular drain is required, this fibrinolytic state is not sufficient to decrease the requirement for ventriculoperitoneal shunt. PMID- 9390697 TI - Regional differences in spectral EEG measures between healthy term and preterm infants. AB - State-specific spectral electroencephalographic (EEG) values were compared among 14 bipolar channel derivations between two healthy neonatal cohorts. Fifty-five healthy preterm neonates of < or = 32 weeks gestational age at birth were studied with 24-channel recordings over 3 hours at term conceptional age. These were compared with studies of 45 healthy term neonates. Five spectral measures for each channel (i.e., total spectral EEG, delta, theta, alpha, and beta frequency ranges) were calculated for each minute, which was identified as active or quiet sleep, based on visual analysis. Using multivariate analysis of variance, differences at each channel were assessed between neonatal cohorts for both states and cohorts; higher total EEG spectral values were noted during active sleep; whereas higher delta and theta spectral values were noted during quiet sleep. The term cohort had higher values for spectral theta, alpha, and beta power spectra in multiple channels, most significantly in the left central (i.e., C3O1) and sagittal regions (FzCz, CzPz) during both states (P < .0001, adj r2 > or = .2). Both interhemispheric and intrahemispheric differences in spectral values were present. For a healthy preterm cohort, lower spectral energies are expressed during sleep in specific head regions. Physiologic asymmetries exist in the newborn brain which are unique for the preterm infant, emphasizing functional alterations in brain development. How these asymmetries are altered by prenatal or postnatal stress or disease states needs to be explored. PMID- 9390698 TI - West syndrome: cerebrospinal fluid nerve growth factor and effect of ACTH. AB - West syndrome is a strictly age-limited encephalopathy of early infancy with unknown pathogenesis. It is often progressive, leading to mental retardation. Neurotrophic factors are important for the regulation of neuronal survival and differentiation, and their expression is influenced by hormones. Levels of beta nerve growth factor in the cerebrospinal fluid were examined by two-site enzyme linked immunosorbent assay method. Human antigen was used as a standard. We present data on largely normal levels of nerve growth factor in the cerebrospinal fluid of infants with cryptogenic etiology, but low or negligible levels in infants with symptomatic etiology, and very high levels in infants with symptomatic postinfectious etiology. Treatment with ACTH led to a greater increase in patients with a good response than in those with a poor response. Low nerve growth factor in patients with symptomatic infantile spasms possibly reflects massive neuronal death. The regression seen in these infants and their poor response to ACTH therapy may be due in part to lack of growth factors supporting neuron survival. This study, previously only demonstrated in animal models, is the first to depict nerve growth factor gene activity in humans as modulated by steroids. PMID- 9390699 TI - Chronic oral VP-16 for recurrent medulloblastoma. AB - Chronic oral VP-16 (Etoposide) is a chemotherapy regimen with wide application in oncology and documented efficacy against germ cell tumors, lymphomas, Kaposi sarcoma, and glial brain tumors. Eight patients ranging in age from 4 to 36 years (median 7.5 years) with locally recurrent medulloblastoma were treated with VP 16. No patient displayed evidence of cerebrospinal fluid dissemination, distant brain or spine parenchymal metastases, or extraneural metastatic disease. All patients had previously been treated with surgery (gross total resection, 5; subtotal resection, 3), craniospinal radiotherapy, and platinum-based chemotherapy (adjuvant, 3; salvage, 8). Each cycle of therapy consisted of 21 days of VP-16 (50 mg/m2/day) followed by a 7 to 14 day rest followed by an additional 21 days of VP-16 (50 mg/m2/day). Complete blood counts were obtained weekly. Neurologic examination and brain magnetic resonance imaging scan with contrast were performed prior to each cycle of therapy. Treatment-related complications included: partial alopecia (5 patients); diarrhea (4); weight loss (3); anemia (2); neutropenia (4); and thrombocytopenia (4). Two patients required transfusion and 1 patient received antibiotics for neutropenic fever. All patients were evaluable for response: 3 demonstrated progressive disease after the first cycle of VP-16, 3 had stable disease (range 4 to 6 months) and 2 had partial neuroradiographic responses (8 and 10 months). Median duration of response and stable disease was 6 months (range: 4 to 10 months) in 5 of 8 (62.5%) patients. Chronic oral VP-16 is a well-tolerated and relatively non-toxic chemotherapeutic agent with demonstrated activity in locally recurrent medulloblastoma. PMID- 9390700 TI - Early expression of proteolipid protein in human fetal and infantile cerebri. AB - Proteolipid protein (PLP) is the major myelin protein of the central nervous system and is widely believed to play an important structural role in maintaining the myelin compaction. We have studied the early developmental changes of PLP with immunohistochemical methods. Our data demonstrate for the first time a comparable scheme for the development of PLP during myelination in human fetal and infant cerebrum. Expression of PLP was first detected in the pallidothalamic fibers and globus pallidus at 20 weeks; it then extended to the striatum at 28 weeks, pericentral gyri and optic radiation at 35 weeks, and acoustic radiation at 40 weeks. Compared to the expression of myelin basic protein (MBP), another major myelin protein in the central nervous system, the developmental changes of PLP is in the same order as MBP, but the PLP immunoreactivity revealed greater and earlier appearance in the cerebrum than that of MBP in the fetal period. These results imply that PLP is a sensitive and useful marker for early myelination and its disorders. PMID- 9390701 TI - Neurodevelopmental outcome in very low birth weight infants at 24 months and 5 to 7 years of age: changing diagnosis. AB - We describe the long-term development of 53 very low birth weight premature infants. The children were divided into 2 groups on the basis of ultrasound scan, and classified as: group I, patients with normal ultrasound scan or with uncomplicated hemorrhage; and group II, patients with complicated hemorrhage or only parenchymal lesions. Minor and major sequelae detected at 2 years of age were compared with those observed at 5 to 7 years. Our study confirms that most severely handicapped children are identified by age 2 years. Minor sequelae are more evident at 5 to 7 years and subjects with good outcome, as expressed by a McCarthy General Cognitive Index score > 80, present a discordant cognitive profile with verbal scores higher than performance scores. Therefore, we emphasize the importance of follow-up of very low birth weight premature infants until school age and stress that neonatal ultrasound scan diagnosis of parenchymal damage represents an important diagnostic tool in terms of both short and long-term neurodevelopmental outcome. PMID- 9390702 TI - Ragged-red fibers and complex I deficiency in a neonate with arthrogryposis congenita. AB - We describe a neonate with hypotonia, weakness, early death owing to respiratory failure, and a severe form of arthrogryposis multiplex congenita. Postmortem studies revealed numerous ragged-red fibers and central nervous system abnormalities consistent with a mitochondrial disease. No NADH:ubiquinone-1 oxidoreductase (complex I) activity could be detected in skeletal muscle. These findings suggest that mitochondrial cytopathies can be associated with arthrogryposis multiplex congenita and should therefore be sought in neonates presenting with severe arthrogryposis. PMID- 9390703 TI - Regional cortical dysplasia associated with suspected hypomelanosis of Ito. AB - A 15-year-old girl with epilepsy, whose skin lesions were reminiscent of hypomelanosis of Ito, is reported. She manifested hypopigmented linear streaks on her upper and lower limbs. Brain magnetic resonance imaging examinations demonstrated poor differentiation of cerebral gray and white matter of her left occipital lobe, with accompanying gliosis. This region also revealed narrowing of sulci, considered to be mass effect. In this region, almost continuous spike discharges were evident on electroencephalograms, and low-perfusion status was observed on single photon emission computed tomography at rest. She also manifested right lower homonymous quadrant anopsia, which may have its origin in the lesion detected, which appeared to be a migration disorder of neuroblasts in our patient, suggesting that the spectrum of hypomelanosis of Ito might be involved. PMID- 9390704 TI - Early neuroradiologic evidence of degeneration in Menkes' disease. AB - We report the case of an infant with Menkes' disease who presented with moderately abnormal neurologic findings at birth and with extraaxial bleeding in the posterior fossa. Early cerebellar atrophy and hypomyelination were evident on magnetic resonance imaging at 5 weeks of age. We suggest that neurodegeneration occurs earlier than has been previously reported and may be identifiable even in neonates. PMID- 9390705 TI - Echocardiographic sign for cerebral ischemia. AB - We describe a newborn who underwent balloon dilatation of critical aortic stenosis and surgical correction of coarctation of the aorta. Postoperative Doppler echocardiogram revealed diastolic retrograde flow in the distal aortic arch and increased systolic flow as well as diastolic forward flow in the left common carotid artery. Cranial computed tomography suggested increased left middle cerebral artery flow and a subacute infarction with luxury perfusion and damage to the blood brain barrier. Therefore, in the absence of another reason for diastolic "run-off" in the distal aortic arch, this flow pattern may represent the echocardiographic sign for cerebral hyperemia associated with subacute ischemia. PMID- 9390706 TI - Enhanced magnetic resonance imaging of leptomeningeal angiomatosis. AB - We present two patients with unilateral occipital gyriform calcification and seizures. Gyriform or serpentine calcification as revealed by computed tomography (CT) scan is rare and is a characteristic finding of Sturge-Weber syndrome (SWS) and celiac disease (CD). These patients had neither the facial nevus flammeus or neurological deficits characteristic of SWS, nor the gastrointestinal symptoms characteristic of CD. CD is often accompanied by cerebral occipital calcification indistinguishable from that of SWS. We demonstrate the presence of cerebral leptomeningeal angiomatosis (LA) by Gadolinium-DTPA-enhanced magnetic resonance imaging (MRI) but could not detect LA by either CT scanning or angiography. It has been reported that contrast-enhanced MRI is useful to detect LA in SWS. However, we found no reports of enhanced MRI in patients with SWS without facial angioma. If future studies can demonstrate the absence of cortical enhancement by contrast-enhanced MRI in CD with cerebral calcifications, enhanced MRI would become an important tool for differentiating CD from SWS. PMID- 9390707 TI - A case of Noonan syndrome with cortical dysplasia. AB - We report the case of a 20-year-old woman with Noonan syndrome. She had severe mental retardation and intractable epilepsy. Magnetic resonance imaging revealed dilated perivascular spaces and a dysplastic lesion in the left temporal lobe, which is thought to have caused her neurologic symptoms. These findings suggest that neuronal in addition to somatic migration can be impaired in Noonan syndrome. PMID- 9390708 TI - Acute hydrocephalus following carbon monoxide poisoning. AB - Carbon monoxide remains a significant cause of poisoning in children. Cerebral edema is often the cause of significant morbidity and mortality in exposed children. While lesions of the basal ganglia have been well documented, the advent of neuroimaging has allowed antemortem demonstration of infarctions of the globus pallidus and putamen with carbon monoxide intoxication. Acute hydrocephalus following carbon monoxide poisoning has been a rare occurrence. We report a 2 year 6 month-old boy who, to our knowledge, represents the first reported case in which repeat computed tomography documented the evolution of hydrocephalus due to carbon monoxide exposure in a child. PMID- 9390709 TI - Purification and gas chromatographic-mass spectrometric characterization of non methylene interrupted fatty acid incorporated in rat liver. AB - A C20 non-methylene interrupted trienoic acid detected in the liver of rat fed with a pine (Pinus koraiensis) seed oil diet was purified by two-step argentation thin-layer chromatography (AgTLC) and characterized by gas chromatography-mass spectrometry (GC-MS). First, a C20 methyl trienoate fraction was obtained from fatty acid methyl esters prepared from rat liver by 5% AgTLC developed with petroleum ether-diethyl ether-acetic acid (70:20:2, v/v) as a solvent system. The fraction was then subjected to AgTLC developed with benzene-acetone-diethyl ether acetic acid (65:15:15:5, v/v) which could separate non-methylene interrupted fatty acids (NMIFA) from usual MIFAs. The purified C20 NMIFA was partially hydrogenated, and the resulting three kinds of the C20 monoenoate were analyzed by GC-MS after conversion to their dimethyl disulfide (DMDS) adducts. The results revealed that the original C20 non-methylene interrupted trienoic acid detected in the liver of rats fed with a pine seed oil diet was delta-5,11,14/20:3, a minor component of pine seed oil. PMID- 9390710 TI - Quantitative analysis of exogenous peptides in plasma using immobilized enzyme cleavage and gas chromatography-mass spectrometry with negative ion chemical ionization. AB - A method is presented for the analysis of peptides in plasma at picomole to femtomole levels. Peptides are isolated from plasma by solid-phase extraction, the peptide of interest is purified by reversed-phase high-performance liquid chromatography (HPLC) and selectively digested using immobilized trypsin or chymotrypsin to yield specific di- or tripeptides. These di- and tripeptides are esterified using heptafluorobutyric anhydride, alkylated with pentafluorobenzyl bromide, then quantified by gas chromatography-mass spectrometry with negative ion chemical ionization. This method has been evaluated for a model synthetic heptapeptide, using a deuterium labeled analog as an internal standard. The half life of the heptapeptide in human plasma was found to be 2 min. Extraction efficiencies of a tritiated peptide of similar size to the heptapeptide, [3H]DSLET, from plasma using either C18 or strong cation-exchange columns were 85+/-3 and 70+/-2%, respectively. Quantitation of fragments from the heptapeptide indicated that the analysis was linear from 1-50 ng of the heptapeptide per ml of plasma. This method was subsequently employed for pharmacokinetic studies of the biologically active peptide Met-enkephalin-Arg-Gly-Leu, where linearity was obtained from 50 to 1000 ng/ml in rat plasma. This method demonstrated negligible side reaction by-products due to autolysis, and has potential for extensive use given the wide availability of gas chromatography-mass spectrometry. PMID- 9390711 TI - Simultaneous determination of polyamines, N-acetylated polyamines and the polyamine analogues BE-3-3-3 and BE-4-4-4-4 by capillary gas chromatography with nitrogen-phosphorus detection. AB - We describe a method for the profiling of polyamines, N-acetylated polyamines and the polyamine analogues N1,N11-bis(ethyl)norspermine (BE-3-3-3) and 1,19 bis(ethylamino)-5,10,15-triazanonadecane (BE-4-4-4-4) in L1210 murine leukaemia cells by capillary gas chromatography with nitrogen-phosphorus detection. The method makes use of four intemal standards. Prepurification comprises deproteinization, isolation with Sep-Pak silica at pH 9.0, conversion to heptafluorobutyryl derivatives and postderivatization organic fluid extraction. Within- and between-series precisions (given as CV.s) for analysis of 1-2x10(6) cells were: putrescine 5.5 and 29.4%; spermidine 1.6 and 7.1%; and spermine 3.2 and 7.6%, respectively. Recoveries relative to the respective internal standards, were in the 70.6-104.7% range. Accuracy and precision of measurements of BE-4-4-4 4 can probably be improved by the introduction of a separate pentamine internal standard. We conclude that the method can be used for studying the effect of BE-3 3-3 and BE-4-4-4-4, and possibly their metabolites, on polyamine homeostasis (biosynthesis, retroconversion, transport, terminal catabolism) and polyamine function. PMID- 9390712 TI - Determination of endogenous levels of 13-cis-retinoic acid (isotretinoin), all trans-retinoic acid (tretinoin) and their 4-oxo metabolites in human and animal plasma by high-performance liquid chromatography with automated column switching and ultraviolet detection. AB - A highly sensitive HPLC method with automated column switching was developed for the simultaneous determination of endogenous levels of 13-cis-retinoic acid (isotretinoin), all-trans-retinoic acid (tretinoin) and their 4-oxo metabolites in plasma samples from man, Cynomolgus monkey, rabbit, rat and mouse. Plasma (0.4 ml) was deproteinated by adding ethanol (1.5 ml) containing the internal standard acitretin. After centrifugation, 1.4 ml of the supernatant were directly injected onto the precolumn packed with LiChrospher 100 RP-18 (5 microm). 1.25% ammonium acetate and acetic acid-ethanol (8:2, v/v) was used as mobile phase during injection and 1% ammonium acetate and 2% acetic acid-ethanol (102:4, v/v) was added, on-line, to decrease the elution strength of the injection solution. After backflush purging of the precolumn, the retained components were transferred to the analytical column in the backflush mode, separated by gradient elution and detected at 360 nm. Two coupled Superspher 100 RP-18 endcapped columns (both 250x4 mm) were used for the separation, together with a mobile phase consisting of acetonitrile-water-10% ammonium acetate-acetic acid: (A) 600:300:60:10 (v/v/v/v), (B) 950:20:5:20 (v/v/v/v), and (C) 990:5:0:5 (v/v/v/v). The method was linear in the range 0.3-100 ng/ml, at least, with a quantification limit of 0.3 ng/ml. The mean recoveries from human plasma were 93.2%-94.4% and the mean inter assay precision was 2.8%-3.2% (range 0.3-100 ng/ml). Similar results were obtained for animal plasma. The analytes were found to be stable in the plasma of all investigated species stored at -20 degrees C for 4.3 months and at -80 degrees C for 9 months, at least. At this temperature, human plasma samples were even stable for 2 years. The method was successfully applied to more than 6000 human and 1000 animal plasma samples from clinical and toxicokinetic studies. Endogenous levels determined in control patients and pregnant women were similar to published data from volunteers. PMID- 9390713 TI - Combination of high-performance liquid chromatography and thin-layer chromatography separation of five adducted nucleotides isolated from liver resulting from intraperitoneal administration with 7H-dibenzo[c,g]carbazole to mice. AB - 7H-Dibenzo[c,g]carbazole, DBC, is a potent environmental liver carcinogen. Liver DNA from mice treated with DBC exhibited seven distinct DBC-DNA adducts as detected by 32P-postlabeling using multidimensional TLC. To improve quantitation and chemically characterize the adducts, DNA samples were hydrolyzed, 32P postlabeled and the adducts were separated from the unadducted normal nucleotides on TLC using a D1 solvent, 0.65 M sodium phosphate (pH 6.8). Adducts were eluted from the TLC plates with 4.0 M pyridinium formate, concentrated, resuspended in 50% aqueous methanol and injected onto the HPLC; five individual adduct peaks were resolved and collected by this method. This approach will prove useful to decrease analysis time and improve chemical characterization of tightly clustered DNA adducts generated in vivo. PMID- 9390714 TI - Bile acid kinetics in man studied by radio thin-layer chromatography and densitometry coupling. AB - A method based on coupling of the techniques of radioscanning a TLC plate and densitometry has been developed for the determination of pool sizes and fractional turnover rate of bile acids in man after intraduodenal administration of 14C-labelled acid. The validity of the method has been checked by comparison of the results obtained with those of an enzymatic spectrophotometric analysis, and a measurement of the radioactivity by liquid scintillation counting, after elution of the separated bile acid from a TLC plate. Advantages of the proposed method over the previous one include a reduced number of manipulations, the possibility of automation, a better reproducibility, and the possibility of elaborating the radiometric data obtained for the primary bile acid for better characterising its metabolism inside the enterohepatic circulation. PMID- 9390715 TI - Capillary electrophoresis of glutamate and aspartate in rat brain dialysate. Improvements in detection and analysis time using cyclodextrins. AB - Addition of cyclodextrins (CDs) to the electrolyte buffer in the capillary zone electrophoresis (CZE) separation of derivatized amino acids was evaluated in terms of fluorescence signal enhancement, resolution, and migration time effects. Maximum fluorescence signal enhancement was observed with separation buffers containing 4 mM beta-cyclodextrin or 10 mM hydroxypropyl beta-cyclodextrin. Resolution values decreased as the CD concentrations increased. Migration times were dependent on CD concentration. Inclusion complex formation constants calculated using changes in migration time showed slight agreement with those calculated by the steady-state fluorescence enhancement technique. Analysis of 20 microl of rat brain microdialysate by CZE using 4 mM beta-cyclodextrin in borate buffer resulted in baseline resolution of glutamate and aspartate in 3.6 min. The results of this work indicate that, when used as separation buffer additives, cyclodextrins are capable of increasing the fluorescence signal and decreasing the migration times of NDA-derivatized acidic amino acids. PMID- 9390716 TI - Simultaneous determination of amphetamine and its analogs in human whole blood by gas chromatography-mass spectrometry. AB - A sensitive and specific gas chromatography-mass spectrometry (GC-MS) method for the determination of amphetamine (AM), methamphetamine (MA), methylenedioxyamphetamine (MDA), methylenedioxymethamphetamine (MDMA) and methylenedioxyethylamphetamine (MDEA) in whole blood was designed, using the respective pentadeuterated analogs of the analytes as internal standards (I.S.). After alkalinisation of blood samples, the amphetamines were extracted using diethyl ether, derivatized with heptafluorobutyric anhydride, then purified by successive washings with deionized water and 4% NH4OH. Extraction recoveries were 85.2% for AM, 90.9% for MA, 76.5% for MDA, 84.1% for MDMA and 63.6% for MDEA. Chromatographic separation was performed on a non-polar 30 m x 0.32 mm HP 5 MS capillary column using a temperature program. Detection was carried out in the electron-impact, selected ion-monitoring mode, using three mass-to-charge ratios for each analyte and one for each I.S. Limits of detection ranged from 0.5 to 8 ng/ml and limits of quantification were 10 ng/ml for AM, MDMA and MDEA; 20 ng/ml for MA; and 50 ng/ml for MDA. The method was linear from this limit up to 1000 ng/ml for all analytes, with good intra-assay precision and good intermediate precision and accuracy over these ranges. There was no interferences from other sympathomimetic drugs such as ephedrine, norephedrine or methoxyphenamine. This method is thus suitable for clinical and forensic toxicology, as well as for doping control. PMID- 9390717 TI - Hair analysis for drugs of abuse. XIX. Determination of ephedrine and its homologs in rat hair and human hair. AB - A sensitive GC-MS method was developed for the quantitative analysis of ephedrine (EP), phenylpropanolamine (PPA) and methylephedrine (ME) in animal and human hair. After washing with 0.1% sodium dodecyl sulfate, hair samples (10 mg) were added with deuterated internal standards, extracted by 1-h sonication and over night soaking in 2 ml of 5 M HCl-methanol (1:20) at room temperature. Following evaporation of the liquid phase, the residue was dissolved in phosphate buffer solution (pH 6.0) and purified using a solid-phase extraction procedure with Bond Elut Certify columns. Two types of derivatization were compared - using trifluoroacetic anhydride (TFAA) and pentafluoropropionic anhydride (PFPA) - for discrimination of EP and methamphetamine (MA). Derivatized extracts were analyzed by GC-MS in the EI mode using a capillary column (OV-1 equivalent). From the results comparing three GC-MS conditions, PFP-derivatives separated with a temperature gradient of 20 degrees C/min from 60 degrees C to 280 degrees C gave the best resolution between EP and MA. ME was analyzed as a trimethylsilyl derivative using N,O-bis-trimethylsilyl acetamide at the above GC condition. The assay was linear from 0.5 to 50 ng/mg (r=0.998) and capable of detecting less than 50 pg of derivatized EP, PPA and ME on-column. Intra-assay precision was characterized by C.V. values from 5 to 16% in the concentration range of 1-10 ng/mg hair. The method was used for the quantitative determination of EP, PPA and ME in the hair obtained from three rats with dark brown hair after ten intraperitoneal injections (5 mg/kg/day) of the three drugs and from three male and one female volunteers with black hair after an oral dose of 50 mg/day of EP HCl for three days. Hair samples were collected by shaving from the back of rats and cutting from the scalp of humans 28 days after the first dose. The incorporation rates of EP, PPA and ME into hair (the ratios of [hair concentration] to [AUC]) obtained from the animal experiment were 0.10, 0.07 and 0.03, respectively, which are a little lower than those (0.14, 0.10 and 0.04) of their desoxy-compounds, MA, amphetamine and dimethylamphetamine. EP was detected at an average of 2.25 ng/mg (n=4) in human scalp hair and at a range of 1-29 ng/mg (n=3) in human beard hair until day 14, but its metabolite (PPA) was at a trace level in the hair of the four subjects. The method was successfully used for detection of ME and EP in the hair of a neonate and its mother who was abusing Bron syrup containing ME during the pregnancy. PMID- 9390718 TI - Determination of low levels of poly(ethylene glycol) 400 in plasma and urine by capillary gas chromatography-selected ion-monitoring mass spectrometry after solid-phase extraction. AB - A convenient and sensitive method for the quantitative determination of poly(ethylene glycol) 400 in plasma and urine with capillary gas chromatography mass spectrometry has been developed. The sample preparation involves solid-phase extraction with subsequent derivatization with heptafluorobutyric anhydride, which proved to give the most stable derivative. The derivatization procedure was optimized using experimental design, and different solid-phase extraction columns were evaluated. The limit of quantitation was 1 micromol/l (0.4 microg/ml) for both plasma and urine. PMID- 9390719 TI - High-performance liquid chromatography of unmodified rosin and its applications in contact dermatology. AB - Rosin is a well recognised skin sensitiser and is also amongst the most common causes of occupational asthma. Due to its complex chemical composition, it is difficult to isolate its many components and this has hindered progress in the identification of the specific respiratory and contact allergens it contains. This paper reports the application of high-performance liquid chromatography and other analytical techniques to the isolation and identification of contact allergens in complex mixtures such as rosin. HPLC methods were developed in order to isolate as many rosin components as possible and these were then patch tested on rosin sensitive individuals. The structure of the most dermatologically active component was then determined using mass spectrometry, nuclear magnetic resonance and infrared techniques. An HPLC method has also been developed which will enable the identification of rosin in commercial products, providing a valuable tool for determining the cause of rosin contact allergy. Furthermore, mass spectral data for the common abieitic-type resin acids are compiled which were used to confirm the identification of the HPLC resin acid peaks and have not been reported previously. PMID- 9390720 TI - Analysis of a novel antiinflammatory agent, 1-(7-tert.-butyl-2,3-dihydro-3,3 dimethylbenzo[b]furan-5-yl)-4- cyclopylbutan-1-one (PGV-20229), in plasma matrices by stable-isotope-dilution gas chromatography-mass spectrometry. AB - A sensitive and selective GC-MS method was developed for the determination of low levels of a novel antiinflammatory agent, 1-(7-tert.-butyl-2,3-dihydro-3,3 dimethylbenzo[b]furan-5-yl)-4- cyclopropylbutan-1-one (I), in small volumes of animal plasma. The method involved the addition of 13C6-labeled-I to plasma samples, followed by a simple liquid-liquid extraction with hexane to isolate the analytes from matrix components. The levels of I in the sample extracts were determined by isotope-dilution GC-MS analysis using selected-ion monitoring. The method was linear over three orders of magnitude, with a limit of quantitation of 1.8 ng/ml I, using plasma sample volumes of 0.1 ml. The method was utilized to determine the pharmacokinetic parameters of I in rats and dogs, following intravenous administration. PMID- 9390721 TI - Screening for forensically relevant benzodiazepines in human hair by gas chromatography-negative ion chemical ionization-mass spectrometry. AB - A procedure is presented for the detection in human hair of forensically relevant benzodiazepines, i.e. nordiazepam, oxazepam, bromazepam, diazepam, lorazepam, flunitrazepam, alprazolam and triazolam. The method involves decontamination of hair with methylene chloride, pulverization in a ball mill, incubation of 50 mg powdered hair in Soerensen buffer (pH 7.6) in the presence of prazepam-d5 used as internal standard, liquid-liquid extraction with diethyl ether-chloroform (80:20, v/v) and gas chromatography-mass spectrometry using negative chemical ionization after derivatization with N,O-bis(trimethylsilyl)trifluoroacetamide plus 1% trimethylchlorosilane. The limits of detection for all benzodiazepines ranged from 1 to 20 pg/mg using a 50-mg hair sample. Coefficients of variation and extraction recoveries, ranging from 7.4 to 25.4% and 47.6 to 90%, respectively, were found suitable for a screening procedure. One hundred and fifteen samples were submitted to this screening procedure, and specimens tested positive for nordiazepam (0.20-18.87 ng/mg, n=42) and its major metabolite oxazepam (0.10-0.50 ng/mg, n=14), flunitrazepam (19-148 pg/mg, n=31), lorazepam (31-49 pg/mg, n=4) and alprazolam (0.3-1.24 ng/mg, n=2). Bromazepam, diazepam and triazolam were not detected. PMID- 9390722 TI - Validated assay for the quantification of anastrozole in human plasma by capillary gas chromatography-63Ni electron capture detection. AB - An assay was developed for the quantification of anastrozole [2,2'-[5-(1H-1,2,4 triazol-1-ymethyl)-1,3-phenylene]bis(2-++ +methylpropiononitrile)] in human plasma using liquid-liquid extraction. Anastrozole and an internal standard were chromatographed and detected by gas chromatography with electron capture detection, using a combination temperature-pressure program. The range of the assay is 3 to 100 ng/ml. Anastrozole was quantified by comparing its peak area to that of an internal standard. A cross-validation of this assay was also successfully performed between several laboratories. PMID- 9390723 TI - Determination of the substance P receptor antagonist CP-122,721 in plasma by narrow-bore high-performance liquid chromatography-ionspray tandem mass spectrometry. AB - A simple, highly sensitive and specific LC-MS-MS assay was developed for the determination of CP-122,721 (I) in rat and human plasma. I and a structural analog, CP-129,943 (II, internal standard), were extracted from plasma with methyl tert.-butyl ether (MTBE). The dried MTBE extracts were reconstituted and analyzed using a narrow-bore (2.1 mm I.D.) YMC basic HPLC column and a mobile phase of acetonitrile-20 mM ammonium acetate, pH 5 (50:50, v/v). Column effluents were monitored by ionspray tandem mass spectrometry. Multiple reaction monitoring (MRM) using the parent to product ion combinations of m/z 381-->205 and 395-->219 was used to quantitate I and II, respectively. The assay exhibited a linear dynamic range of 0.2-100 ng/ml. Absolute recoveries from plasma were above 80% for both I and II. The precision and accuracy values for the method were within +/-3 and +/-9%, respectively. Sample analysis times were less than 5 min from one injection to the next. The assay has proved to be applicable to the pharmacokinetic study of I in rats. PMID- 9390724 TI - Simultaneous quantification of cefotaxime, desacetylcefotaxime, ofloxacine and ciprofloxacine in ocular aqueous humor and in plasma by high-performance liquid chromatography. AB - Cefotaxime, given intravenously, is currently used as a broad-spectrum antibiotic for prophylaxis of intra- and postoperative infections in ocular lens surgery. A proposed therapeutic and economic alternative is the use of orally active fluoroquinolone ofloxacine as prophylactic agent. A HPLC method was developed for determination of both antibiotics in ocular aqueous humor and plasma in order to optimize dosage for safe surpassing minimal inhibitory concentration in the humor compartment. For plasma determinations a solid-phase extraction procedure was used with ciprofloxacine as internal standard. Detection limits for direct HPLC analysis of ocular aqueous humor was 0.08 microg/ml for all compounds, whereas in plasma 0.31 microg/ml could be determined after solid-phase extraction. PMID- 9390725 TI - Enantioselective high-performance liquid chromatography determination of methadone enantiomers and its major metabolite in human biological fluids using a new derivatized cyclodextrin-bonded phase. AB - The simultaneous determination of methadone (Mtd) enantiomers and its major metabolite, 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP), in human urine and serum by enantioselective HPLC using a new Cyclobond 1-2000 RSP column is described. After alkaline extraction from urine or serum with estazolam as an internal standard, Mtd enantiomers and its metabolite (EDDP) are separated on the previous column with reversed-mobile phase and detected at 210 nm. Peak resolutions are about 2.0 for Mtd enantiomers. The relative standard deviations (R.S.D.) of Mtd and EDDP standards are between 0.5 and 4.5%. Most drugs of abuse are shown not to interfere with this technique. The method has been applied to study the levels of each Mtd enantiomer and of its racemic metabolite in urine and serum of patients under maintenance treatment for opiate dependence. In urine, R-(-)-Mtd levels are always higher (about 2+/-0.5-fold) than those of S (+)-Mtd and in most cases, metabolite concentrations are greater than those of global Mtd enantiomers. However, the R-(-) enantiomer levels of residual drug in serum of some patients were lower than those of its antipode. This method is suitable for pharmacokinetic and toxicological studies of Mtd enantiomers and its major metabolite in biological fluids. PMID- 9390726 TI - Stereoselective determination of R-(+)- and S-(-)-remoxipride, a dopamine D2 receptor antagonist, in human plasma by chiral high-performance liquid chromatography. AB - A stereoselective high-performance liquid chromatographic (HPLC) method is described for the selective and sensitive quantitation in human plasma of R-(+)- and S-(-)-enantiomers of remoxipride. Remoxipride was extracted from basified plasma into hexane-methyl-tert.-butyl ether (20:80, v/v), washed with sodium hydroxide (1.0 M), then back-extracted into phosphoric acid (0.1 M). A structural analog of remoxipride was used as an internal standard. The sample extracts were chromatographed using a silica-based derivatized cellulose chiral column, Chiralcel OD-R, and a reversed-phase eluent containing 30-32% acetonitrile in 0.1 M potassium hexafluorophosphate. Ultraviolet (UV) absorbance detection was performed at 214 nm. Using 0.5-ml plasma aliquots, the method was validated in the concentration range 0.02-2.0 microg/ml and was applied in the investigation of systemic inversion of remoxipride enantiomers in man. PMID- 9390727 TI - Quantitation of trimipramine enantiomers in human serum by enantioselective high performance liquid chromatography and mixed-mode disc solid-phase extraction. AB - A sensitive and stereospecific method for the quantitation of trimipramine enantiomers in human serum was developed. The assay involves the use of a novel mixed-mode disc solid-phase extraction for serum sample clean-up prior to HPLC analysis and is also free of interference from the enantiomers of desmethyltrimipramine, 2-hydroxytrimipramine, and 2-hydroxydesmethyltrimipramine, the three major metabolites of trimipramine. Chromatographic resolution of trimipramine enantiomers was performed on a reversed-phase cellulose-based chiral column (Chiralcel OD-R) under isocratic conditions using a mobile phase consisting of 0.3 M aqueous sodium perchlorate-acetonitrile (58:42, v/v) at a flow-rate of 0.5 ml/min. Recoveries for R- and S-trimipramine enantiomers were in the range of 93-96% at 25-185 ng/ml levels. Intra-day and inter-day precisions calculated as R.S.D. were in the ranges of 0.30-8.00% and 1.60-10.20% for both enantiomers, respectively. Intra-day and inter-day accuracies calculated as percent error were in the 0.01-2.10% and 1.00-3.00% ranges for both enantiomers, respectively. Linear calibration curves were in the concentration range 15-250 ng/ml for each enantiomer in serum. The limit of quantification of each enantiomer was 15 ng/ml. The detection limit for each enantiomer in serum using a UV detector set at 210 nm was 10 ng/ml (S/N=2). In addition, separation of the enantiomers of desmethyltrimipramine, 2-hydroxytrimipramine, and 2 hydroxydesmethyltrimipramine were investigated. The desmethyltrimipramine enantiomers could be resolved on the Chiralcel OD-R column under the same chromatographic conditions as the trimipramine enantiomers, but the other two metabolite enantiomers required different mobile phases on the Chiralcel OD-R column to achieve satisfactory resolution with Rs values of 1.00. PMID- 9390728 TI - Sensitive and rapid method for the simultaneous quantification of five antidepressants with their respective metabolites in plasma using high performance liquid chromatography with diode-array detection. AB - A high-performance liquid chromatographic method with diode array detection (HPLC DAD) has been developed for the simultaneous separation and quantification of imipramine, amitriptyline, maprotiline, fluoxetine, clomipramine and their respective metabolites using a 500-microl plasma sample and clovoxamine as the internal standard. The substances were eluted on a Symmetry C18 5-microm column (250x4.6 mm, I.D.). Full UV spectra from 200 to 450 nm were recorded on-line during the entire analysis and were automatically compared to spectra stored in a library. The quantification was performed at 226, 254, and 400 nm. Peak height ratios were linear over a concentration range of 10-3000 ng/ml: the correlation coefficient (r) was better than 0.998 for all substances at each wavelength. Acceptable coefficients of variation are demonstrated for both within-run and day to-day assays. The method is simple, highly specific and currently being used for drug monitoring and toxicological studies in children and adult patients. PMID- 9390729 TI - Determination of SDZ ICM 567 in blood and muscle microdialysis samples by microbore liquid chromatography with ultraviolet and fluorescence detection. AB - Fast, simple and accurate methods for the determination of SDZ ICM 567, the 7 methoxy derivative of tropisetron, in microdialysates have been developed. Sampling by microdialysis from freely moving rats in the portal and jugular vein offers a new technology for pharmacokinetic studies by direct and continuous measurement of unbound drug concentrations with time. SDZ ICM 567 can be identified in small sample volumes of dialysates on a microbore high-performance liquid chromatography column-switching system with ultraviolet detection. In addition, determination of SDZ ICM 567 by fluorimetric detection has been developed for muscle microdialysates from rats. [14C]SDZ ICM 567 was used as reference substance for the estimation of the amount of substance transferred through the dialysis membrane. The radioactive measurement (RA) gave the recovery information, whereas the liquid chromatographic method detected the sum of [14C]SDZ ICM 567 and dialyzed SDZ ICM 567. PMID- 9390730 TI - Determination of propiomazine in rat plasma by direct injection on coupled liquid chromatography columns with electrochemical detection. AB - This method describes the determination of propiomazine by direct injection of rat plasma into a chromatography system based on coupled reversed-phase columns. An extraction column, packed with porous silica particles with covalent-bound alpha1-acid glycoprotein (AGP), was used to separate the plasma proteins from the analyte. After isolation the analyte was transferred to the analytical column for separation and detection. Propiomazine was detected by an electrochemical detector and the limit of quantification was 2.0 ng/ml (100 pg injected). The absolute recovery was 80.9+/-2.4% at 9.0 ng/ml level. The inter-day and intra-day precision was 10.9% (5.6 ng/ml) and 2.8% (9.0 ng/ml), respectively. PMID- 9390731 TI - Determination of 6,4'-bis-(2-imidazolinylhydrazone)-2-phenylimidazo[1,2-a]py ridine in plasma and whole blood by high-performance liquid chromatography. AB - A selective and sensitive HPLC assay for the quantitative determination of a new antifilarial drug, 6,4'-bis-(2-imidazolinylhydrazone)-2-phenylimidazo[1,2-a]pyr idine (CDR 101) is described. After extraction from plasma and blood, CDR 101 was analysed using a C18 Nucleosil ODS column (250x4.6 mm, 5 microm particle size) and mobile phase of acetonitrile-0.05 M ammonium acetate adjusted to pH 3.0, with UV detection at 318 nm. The mean recoveries of CDR 101 in plasma and blood over a concentration range of 25-500 ng/ml were 95.5+/-2.01% and 83.3+/-1.87%, respectively. The within-day and day-to-day coefficient of variations for plasma were 3.23-6.21% and 2.59-9.90%, respectively, those for blood were 2.59-5.92% and 2.89-6.82%, respectively. The minimum detectable concentration for CDR 101 was 1 ng/ml in plasma and 2.5 ng/ml in whole blood. This method was found to be suitable for clinical pharmacokinetic studies. PMID- 9390732 TI - Stereospecific determination of mefloquine in biological fluids by high performance liquid chromatography. AB - A sensitive stereoselective HPLC method was developed for determination of mefloquine (MFQ) enantiomers in plasma, urine and whole blood. The assay involved liquid-liquid extraction of MFQ from biological fluids with a mixture of hexane and isopropanol in the presence of sodium hydroxide and derivatization of the residue by (+)-(S)-naphthylethylisocyanate (NEIC) as chiral derivatizing reagent. Separation of the resulting diastereomers was performed on a silica normal-phase column using chloroform-hexane-methanol (25:74:1) as the mobile phase with a flow rate of 1 ml/min. Using 200 microl of plasma or whole blood, the limit of determination was 0.2 microg/ml with UV detection for both enantiomers. The limit of determination in 500 microl of urine was 0.08 microg/ml with UV detection. PMID- 9390733 TI - Confirmation of malachite green, gentian violet and their leuco analogs in catfish and trout tissue by high-performance liquid chromatography utilizing electrochemistry with ultraviolet-visible diode array detection and fluorescence detection. AB - A sensitive analytical procedure for the confirmation of residues of malachite green (MG), gentian violet (GV) and their leuco analogs (LMG and LGV) in catfish and trout tissue at 10 ng/g is described. Frozen (-20 degrees C) fish fillets were cut into small pieces and homogenized in Waring blendors. The compounds of interest were extracted from 20-g amounts of homogenized fish tissue with acetonitrile-buffer, partitioned against methylene chloride, and isolated with tandem neutral alumina and propylsulfonic acid cation-exchange solid-phase extraction cartridges. Samples of 100 microl (0.8 g equiv.) were chromatographed isocratically in 10 min using an acetonitrile-buffer mobile phase on a short chain deactivated (SCD) reversed-phase column (150x4.6 mm I.D.) in-line with a post-column oxidation coulometric electrochemical cell (EC), a UV-Vis diode array detector and a fluorescence detector. PMID- 9390734 TI - Quantitative comparison on the refinement of horse antivenom by salt fractionation and ion-exchange chromatography. AB - A quantitative comparison was made on the fractionation of pepsin-digested horse antivenoms by ammonium sulfate (AS) fractional precipitation and ion-exchange chromatography on Q-Sepharose. In the precipitation process, pepsin digested horse anti-Naja kaouthia serum was precipitated by 30% saturated AS followed by 50% saturated AS. The recovery of antibody activity [as measured by an enzyme linked immunosorbent assay (ELISA) against the cobra postsynaptic neurotoxin 3] from the 30-50% saturated AS precipitate was 53% with a 1.93-fold purification. For the chromatographic process, the behavior of the horse antitoxin antibody and its F(ab')2 fragments was first studied. The pepsin digested horse serum was then desalted on a Bio-gel P-2 column followed by chromatography on Q-Sepharose using a linear gradient (20 mM Tris-HCl, pH 8.0 containing 0.0 to 0.5 M NaCl) A peak containing primarily the F(ab')2 antibody could be obtained. This peak constituted 73% of the total antivenom activity with 2.08-fold purification. The total recovery of antibody activity by the chromatographic process was 90%. The yield of antibody activity was about 2-fold higher than that reported previously with other fractionation procedures. The implications of these results for the refining of horse therapeutic antivenoms are discussed. PMID- 9390736 TI - Analysis of the antidiabetic drug acarbose by capillary electrophoresis. AB - This study describes the derivatization of the pseudooligosaccharide acarbose and its main metabolite, component 2, with 7-aminonaphthalene-1,3-disulfonic acid (ANDS) in human urine. Their efficient separation was possible by means of capillary zone electrophoresis, using a capillary tube of fused-silica containing 100 mM triethylammonium phosphate buffer, pH 1.5. On column laser-induced fluorescence allowed the detection of the pseudooligosaccharides in human urine in the nanomolar range. With this method, acarbose and component 2 were quantified in human urine after application of 300 mg of acarbose. PMID- 9390735 TI - Characterisation of a chromatographically produced anti-D immunoglobulin product. AB - A chromatographic fractionation method has been developed for the production of a liquid-stable anti-D immunoglobulin product for intravenous and intramuscular use. An immunoglobulin fraction, highly enriched with anti-D immunoglobulins, was isolated by cation-exchange column chromatography and further polished, first by anion-exchange chromatography, followed by an aluminium hydroxide gel treatment. The process includes two specific steps for virus inactivation and removal, namely S/D treatment and nanofiltration. The overall anti-D process yield is about 56%. The final product is stabilised with human albumin and glycine and placed in ready-to-use syringes. The anti-D product was shown to be stable in liquid state for at least 30 months at 4 degrees C. PMID- 9390737 TI - Detection of microbial-derived fatty acids in carious dentin by gas chromatography and gas chromatography-mass spectrometry. AB - The aim of the present study was to analyze the fatty acid content of carious and sound human dentin. Gas chromatography and gas chromatography-mass spectrometry revealed the presence of fatty acids of C10-C18 size in the carious dentin, whereas fatty acids of C16 size were present in minute amounts in three samples of the corresponding sound dentine controls. No fatty acids were detected in the other sound dentin control samples. The source of fatty acids was considered to be microorganisms invading the dentin during the progression of the caries lesion. The presence of bacterial fatty acids in carious dentin may serve as a marker for the pathological process and thus contribute to the understanding of the mechanisms involved. PMID- 9390738 TI - Measurement of oxalate in human plasma ultrafiltrate by ion chromatography. AB - An improved ion chromatographic method for the measurement of oxalate in human plasma ultrafiltrate is described. Ultrafiltration was carried out using an appropriate device and procedure. Centrifugation of 0.5 ml heparin plasma at 4 degrees C for 50 min yielded water-clear ultrafiltrate in amounts allowing replicate measurements of oxalate. The specificity of the method was confirmed. The recovery of oxalate added to plasma was approximately 80%, whereas dilution of plasma, and of an oxalate-containing salt solution, resulted in falsely high values; the mechanism(s) underlying this phenomenon are insufficiently understood at present. The intra-assay precision of the method was assessed and from ten replicates of a pool plasma, the inter-assay precision from ten measurements of the same plasma on different days; the observed ranges of oxalate were 1.32-1.56 (mean 1.42) and 1.42-1.64 (mean 1.53) micromol/l, respectively. In plasma ultrafiltrate of a limited number of healthy volunteers the range of oxalate was 1.18-2.50 micromol/l, thus permitting renal oxalate handling to be studied. PMID- 9390739 TI - Microchromatographic quantitation of hemoglobin A levels in phenotypes of sickle cell-beta(+) thalassemia. AB - The inheritance of the sickle cell gene in combination with a gene for beta(+) thalassemia results in a spectrum of sickle cell-beta(+) thalassemia syndromes with varying levels of hemoglobin A (HbA). Some severe sickle cell-beta(+) thalassemia syndromes have small amounts of HbA, which may be difficult to quantitate in the presence of fetal hemoglobin. A microcolumn chromatographic method, using 0.5 M Tris-acetic acid developers with varying pH values from 9.0 to 6.0, appears to adequately quantitate small amounts of HbA. This method is relatively simple and cheaper than high-performance liquid chromatography, a major consideration in developing countries. PMID- 9390740 TI - Thiopurine methyltransferase activity: new high-performance liquid chromatographic assay conditions. AB - This paper reports changes to our previously published high-performance liquid chromatographic method for the measurement of 6-methylmercaptopurine (6-MMP) in red blood cell lysates. The extraction procedure and chromatographic conditions have been improved and the range of the calibration curves has been modified. The recoveries of 10 and 100 ng ml(-1) 6-MMP were 99.0+/-6.0% and 96.3+/-4.0% respectively and the limit of quantification was lowered to 5 ng ml(-1). This method, which does not require radioactive S-adenosyl-L-methionine, is more sensitive, specific and reproducible and may prove useful for routine determination of thiopurine methyltransferase activity in red blood cells. PMID- 9390741 TI - Simultaneous determination of L-dopa and 3-O-methyldopa in human platelets and plasma using high-performance liquid chromatography with electrochemical detection. AB - Various high-performance liquid chromatographic (HPLC) methods for the determination of plasma levels of L-dopa and of its metabolite, 3-O-methyldopa (3 OMD), have been previously described. In this study, we report a modification of these methods, that enables the assay of these two compounds in platelets and plasma obtained from the same sample of whole blood. Reversed-phase (RP) HPLC with electrochemical (coulometric) detection was used. The within-run and between run coefficients of variations, for the two compounds, were less than 10%, in both platelets and plasma; the detection limits for platelet levels of L-dopa and 3-OMD were 2 and 6 ng/10(9) platelets, respectively. In plasma, the detection limits for L-dopa and 3-OMD were 1 and 3 ng/ml, respectively. The method is rapid and simple. When applied to a population of patients with Parkinson's disease under treatment with L-dopa, this method revealed detectable levels of L-dopa and 3-OMD in the platelets of all patients. The application of this technique may provide new insights into the pharmacokinetics of L-dopa in patients with Parkinson's disease. PMID- 9390742 TI - High-performance thin-layer chromatographic determination of 5-methoxypsoralen in serum from patients. AB - A simple and rapid high-performance thin-layer chromatographic (HPTLC) determination of 5-methoxypsoralen in serum is necessary for the therapeutic survey of patients treated with Puvatherapy (psoralen+UVA). The assay for this biological fluid involves an extraction with heptane-dichloromethane (4:1, v/v). The analytical method is linear from 50 to 250 ng/ml. This assay range is adequate for analysing human serum, as it corresponds to psoralen concentrations measured in serum from patients treated with psoralen and UVA against psoriasis and vitiligo. The limit of detection is 15 ng/ml. The coefficient of variation was less than 7%. PMID- 9390743 TI - Dynamics of syncytium-inducing and non-syncytium-inducing type 1 human immunodeficiency viruses during primary infection. AB - Syncytium-inducing (SI) type 1 human immunodeficiency viruses (HIV-1) replicate faster in vitro and are generally more cytopathic to T cells than non-syncytium inducing (NSI) HIV-1. Early in infection, the virus population typically consists of NSI viruses, with SI viruses appearing later. This is true even when both SI and NSI viruses are transmitted, and when SI viruses dominate the virus population peak seen during primary infection. Here, Phillips's model of HIV dynamics during primary infection (Science 271:497-499) is modified to map the growth trajectories of SI and NSI subpopulations. The model predicts that with certain rate constants, SI viruses may show a more precipitous decline in their numbers during primary infection than NSI viruses, and this may account for the observed dominance of NSI viruses early in infection. PMID- 9390744 TI - Thy/Liv-SCID-Hu mice implanted with human intestine: an in vivo model for investigation of mucosal transmission of HIV. AB - Mucosal transmission is a major route by which individuals become infected with HIV. Investigation into the mechanism by which mucosal transmission of HIV occurs would be greatly facilitated by the development of a small animal model infectible with HIV by the mucosal route. We have previously described a SCID-hu mouse model, in which human thymic and liver tissues are implanted under both kidney capsules (thy/liv-SCID-hu mice), which are populated in the periphery with high numbers of human T cells and that develop disseminated HIV-1 infection after intraperitoneal injection. To expand further the usefulness of the thy/liv-SCID hu mouse as a model for studying mucosal transmission of HIV, thy/liv-SCID-hu mice were subcutaneously implanted with human intestinal tissue in a manner that maintained the lumen. Four months later, the histological appearance of the implanted intestine resembled that of normal human bowel tissue and the lamina propria was populated with human T cells. Six weeks after introduction of HIV into the lumen of the intestinal implant, the mice developed disseminated HIV infection. Scattered HIV-infected cells were detected in the lamina propria of the implant, indicating that HIV infection in these mice was mediated by transmission of the virus across the mucosa of the human intestinal implant. Thus, our modified thy/liv-SCID-hu mice transplanted with human bowel tissue should provide a novel model for investigating mucosal transmission of HIV. PMID- 9390745 TI - Human immunodeficiency virus type 1 glycoprotein 120-specific T lymphocytes provide intermolecular help for anti-CD4 autoantibody production in exposed uninfected subjects. AB - Anti-CD4 antibodies have been documented in about 10-20% of HIV-infected patients. This autoimmune response could be triggered by increased CD4 processing and unveiling of hidden (cryptic) epitopes. Multiple markers of exposure to HIV have been described in exposed uninfected individuals. Here, we investigated the mechanisms underlying the generation of anti-CD4 antibodies in a cohort of 54 seronegative exposed uninfected individuals. We identified anti-CD4 antibodies above normal levels in 16 of 47 (34%) exposed uninfected subjects. The fine specificity of these antibodies was different in this cohort when compared with those found in HIV+ patients. This suggested the possibility of different mechanisms underlying the generation of anti-CD4 antibodies in these two groups. Indeed, in exposed uninfected subjects, we found circulating CD4 T cells specific for gp120, but not for CD4. In contrast, HIV-1-seropositive patients had peripheral blood T cells specific for both molecules. Noncovalent binding of gp120 to soluble CD4 enhanced activation of gp120-specific T lymphocytes in exposed uninfected subjects, but not in HIV+ subjects. Moreover, gp120-specific T cells isolated from exposed uninfected, but not from HIV+, subjects provided help for anti-CD4 antibody production by B cells pulsed with CD4-gp120 complex. We conclude that gp120-specific T cells are present in exposed uninfected individuals, and can provide intermolecular help for anti-CD4 antibody production. This mechanism is distinct from that found in HIV-1-seropositive patients and may play a protective role against HIV-1 infection in vivo. PMID- 9390746 TI - Restricted specificity of anti-V3 antibodies induced in humans by HIV candidate vaccines. AB - We analyzed the fine specificity of anti-V3 antibodies elicited in three different species (human, guinea pig, and macaque) by various HIV candidate vaccines. Following immunization with recombinant canarypox virus expressing gp160MN or with recombinant gp160MN/LAI, this antibody response was shown to be directed against the NH2-terminal region of the V3 loop. Although this response was increased by a prime-boost regimen using immunization with canarypox expressing gp160 followed by an rgp160 boost, its specificity remained restricted mainly to the recognition of this region of the V3 loop. Pepscan analysis of sera confirmed the results obtained by ELISA and allowed the definition of an immunodominant common binding site for these sera located within the sequence NKRKRIHIGPGR. In contrast to these results, a boost with the V3 peptide was shown to broaden the antibody response and pepscan analysis showed that sera from individuals boosted with the V3 synthetic peptide recognize determinants all along the V3 loop. Similar fine specificity of anti-V3 antibodies was obtained in human, guinea pig, and macaque following immunization by a prime-boost regimen using canarypox recombinants expressing gp160 or gp120 and purified rgp160. In contrast, a V3 synthetic peptide boost stimulated the production of antibodies that recognize multiple epitopes within the V3 loop. Because the induction of antibodies that recognize multiple sites in the V3 loop could be of major importance to neutralize different HIV isolates, these results may have implications for the design and selection of HIV candidate vaccines. PMID- 9390747 TI - Outcome of immunization of cynomolgus monkeys with recombinant Semliki Forest virus encoding human immunodeficiency virus type 1 envelope protein and challenge with a high dose of SHIV-4 virus. AB - Infection of macaques with chimeric simian-human immunodeficiency viruses (SHIVs) allows evaluation of HIV-1 envelope vaccines. SHIV-4 is based on SIVmac239 but carries the env, tat, and rev genes of HIV-1IIIB. In this study we used Semliki Forest virus (SFV) RNA vectors to express the envelope protein gp160 of HIV-1IIIB in cynomolgus macaques. Monkeys were immunized four times with recombinant suicide SFV. Whereas two of four monkeys showed T cell-proliferative responses, only one monkey had demonstrable levels of antibodies to HIV-1 gp41 and gp120 as shown by enzyme-linked immunosorbent assay (ELISA) and Western blot. The vaccinated monkeys and four control animals were challenged with 10,000 MID100 (100% minimum infectious doses) of cell-free monkey cell-grown SHIV-4 virus. As demonstrated by virus isolation, all macaques became infected after challenge. All vaccinated monkeys showed an HIV-1-specific anamnestic T cell-proliferative response. Three of four vaccines had developed HIV-1-Env-specific antibodies 2 weeks after challenge whereas none of the four controls showed any detectable immune response at this time point. Furthermore, three of four vaccinated monkeys had no demonstrable viral antigenemia and low viral load as opposed to one of the four naive control animals. PMID- 9390748 TI - Immunogenicity of recombinant vaccinia viruses that display the HIV type 1 envelope glycoprotein on the surface of infectious virions. AB - A chimeric protein, consisting of the extracellular domain of the human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein attached to the transmembrane and cytoplasmic domains of the vaccinia virus B5R glycoprotein, is displayed on the surface of extracellular recombinant vaccinia virus particles, whereas the unmodified full-length HIV-1 glycoprotein is not (Katz E, et al., J Virol 1997;71:3178-3187). Here, we report that rabbits and mice inoculated with recombinant vaccinia viruses that express the chimeric protein developed higher HIV-specific antibody responses than animals inoculated with vaccinia virus that expressed the unmodified HIV glycoprotein. These data suggest that the immunogenicity of recombinant proteins may be enhanced by their presentation on the surface of vaccinia virus particles. PMID- 9390749 TI - Study of spontaneous apoptosis in HIV+ patients: correlation with clinical progression and T cell loss. AB - In vitro spontaneous apoptosis (SA) of lymphocytes was studied in HIV infection to evaluate possible clinical and prognostic correlations, in a transsectional study of 101 individuals in different clinical categories and in a prospective longitudinal study of 18 asymptomatic individuals (mean follow-up, 17.2 months). The rate of SA was higher in HIV+ patients than in healthy controls (p < 0.001) and was higher in patients with AIDS than in the other HIV+ individuals (p < 0.001). It was inversely correlated with the peripheral blood CD4+ (R -0.61; p < 0.001) and CD8+ (R -0.46; p < 0.001) cell numbers. In a group of long-term survivors (LTS), it was significantly lower than in a control group of asymptomatic HIV+ patients with a similar number of circulating CD4+ lymphocytes but a shorter follow-up (p < 0.02). In the five asymptomatic HIV-infected individuals who showed a clinical progression, peaks of SA rates above the normal range before the clinical event were much more frequent than in those who remained asymptomatic (p < 0.0001), even though they were fairly homogeneous as far as CD4+ cell count and viral load were concerned. The median levels of SA rates were moreover correlated with the rate of total T cell loss (R -0.46; p 0.053). This study suggests that evaluation of the SA levels may provide a predictive factor for clinical and immunological progression of HIV-related immunodeficiencies and strengthen the hypothesis for the role of this phenomenon in the pathogenesis of this progression. PMID- 9390750 TI - Downregulation of HLA class I antigen expression in CD4+ T lymphocytes from HIV type 1-infected individuals. AB - The expression of HLA class I antigens is downregulated in CD4+ T cells following in vitro HIV-1 infection. We determined whether the expression of HLA class I antigens is downmodulated in peripheral blood lymphocytes (PBLs) of HIV-1 positive subjects and whether this defect correlates with disease progression. A cohort of 62 HIV-1-seropositive individuals in different stages of disease was studied. Among these, four subjects were evaluated at yearly intervals for 6 years. The expression of HLA class I, HLA class II, and CD38 antigens was analyzed in PBLs and in CD4+ and CD8+ T lymphocyte subpopulations. The percentage of HLA class I-positive cells and the membrane density of HLA class I antigens were significantly lower in PBLs from HIV-1-positive individuals than in PBLs from HIV-negative controls, proportionally decreased with disease progression, and significantly correlated with the decrease in CD4+ T lymphocytes. Furthermore, the percentage of HLA class I-positive cells and the membrane density of HLA class I antigens were significantly lower in CD4+ T lymphocytes from AIDS patients with respect to CD4+ T lymphocytes from HIV-negative controls and to CD8+ T lymphocytes from HIV-negative controls and AIDS patients. By contrast, the expression of HLA class II and CD38 antigens was upregulated in CD4+ and CD8+ T lymphocytes from HIV-1-positive subjects. The defective expression of HLA class I antigens could impair the lysis of HIV-infected CD4+ cells by virus-specific HLA class I-restricted cytotoxic T lymphocytes and contribute to the progression of disease. PMID- 9390751 TI - Inhibition of plating of human T cell leukemia virus type I and syncytium inducing types of human immunodeficiency virus type 1 by polycations. AB - We examined the effects of polycations, namely, diethylaminoethyl-dextran (DEAE dextran) and hexadimethrine bromide (Polybrene), on infection with the retroviruses human T cell leukemia virus types I and II (HTLV-I and HTLV-II) and human immunodeficiency virus type 1 (HIV-1). The plating of vesicular stomatitis virus (VSV) pseudotype bearing envelope antigens of HTLV-I [VSV(HTLV-I)] was inhibited about 2- and 10-fold by treatment with DEAE-dextran and Polybrene, respectively. The formation of HTLV-I viral DNA detected 1 day after infection was also inhibited by these polycations. In contrast, polycations enhanced the plating of the VSV (HTLV-II) pseudotype two- to threefold. The polycations did not affect the plating efficiency of HTLV-I or HTLV-II when added after virus adsorption. Infection of human T cell lines, peripheral blood lymphocytes (PBLs), or brain-derived cells with syncytium-inducing (SI) types of HIV-1 strains (GUN1 and IIIB) was inhibited 3- to 20-fold by polycations. However, infection of PBLs or monocyte-derived macrophages with the macrophage-tropic Ba-L or SF162 strain was enhanced 1.5- to twofold by polycations. On the other hand, syncytium formation in coculture induced by HTLV-I, HTLV-II, or HIV-1 was enhanced two- to threefold unanimously by DEAE-dextran or Polybrene. Although polycations have been used to potentiate human retrovirus adsorption, they inhibited infection of cell-free HTLV-I or SI-type HIV-1 strains. PMID- 9390752 TI - A synthetic peptide corresponding to the carboxy terminus of human immunodeficiency virus type 1 transmembrane glycoprotein induces alterations in the ionic permeability of Xenopus laevis oocytes. AB - The carboxy-terminal 29 amino acids of the human immunodeficiency virus type 1 transmembrane glycoprotein (HIV-1 TM) are referred to as lentivirus lytic peptide 1 (LLP-1). Synthetic peptides corresponding to LLP-1 have been shown to induce cytolysis and to alter the permeability of cultured cells to various small molecules. To address the mechanisms by which LLP-1 induces cytolysis and membrane permeability changes, various concentrations of LLP-1 were incubated with Xenopus laevis oocytes, and two-electrode, voltage-clamp recording measurements were performed. LLP-1 at concentrations of 75 nM and above induced dramatic alterations in the resting membrane potential and ionic permeability of Xenopus oocytes. These concentrations of LLP-1 appeared to induce a major disruption of plasma membrane electrophysiological integrity. In contrast, concentrations of LLP-1 of 20-50 nM induced changes in membrane ionic permeability that mimic changes induced by compounds, such as the bee venom peptide melittin, that are known to form channel-like structures in biological membranes at sublytic concentrations. An analog of LLP-1 with greatly reduced cytolytic activity failed to alter the electrophysiological properties of Xenopus oocytes. Thus, by altering plasma membrane ionic permeability, the carboxy terminus of TM may contribute to cytolysis of HIV-1-infected CD4+ cells. PMID- 9390753 TI - Effects of tapering doses of oral prednisone on viral load among HIV-infected patients with unexplained weight loss. AB - In an exploratory study of virologic and immunomodulatory effects of corticosteroid therapy for wasting syndrome, four HIV-infected adults with recent unexplained weight loss were given tapering doses of prednisone over a 2-month period. Serum neopterin and TNF receptor II levels decreased from baseline after 7 days. An antiretroviral effect was observed initially, peaking on days 14-21 (mean change in HIV-1 branched chain DNA assay on day 21 of -0.52 log10; mean change, from baseline to nadir for each individual, of -0.63 log10); subsequent bDNA levels returned toward baseline as prednisone was tapered. No patient lost weight and there was a mean weight gain of 3.5 kg. Anecdotal reports of corticosteroid benefits in the wasting syndrome may result in part from decreased T cell activation leading to decreased HIV replication, an effect that may be self-limited or that may occur only at higher prednisone doses. Studies involving more targeted immunomodulatory agents for wasting syndrome are warranted. PMID- 9390754 TI - Diversity and distribution of gag and env subtypes among 146 HIV type 1 isolates in Taiwan. PMID- 9390755 TI - Phylogenetic analysis of a simian T lymphotropic virus type I from a hamadryas baboon. PMID- 9390756 TI - Regional and cellular distribution of serotonin 5-hydroxytryptamine2a receptor mRNA in the nucleus accumbens, olfactory tubercle, and caudate putamen of the rat. AB - This paper describes the regional and cellular distribution of serotonin 5 hydroxytryptamine2a (5-HT2a) receptor mRNA in (sub)regions of the rat striatum by using in situ hybridization. Our results indicate that 5-HT2a mRNA is distributed heterogeneously in this brain region. Regional densitometry of autoradiograms from striatal sections hybridized with isotope-labeled cRNA probes showed that mRNA levels were highest in the olfactory tubercle, lower in the nucleus accumbens, and lowest in the caudate-putamen. In the nucleus accumbens, the average mRNA levels in the shell were higher than those in the core. These data suggest a particular relevance for the 5-HT2a receptor for olfactory tubercle- and shell-related functions. Therefore, in the nucleus accumbens and the olfactory tubercle, the cellular localization of 5-HT2a mRNA was investigated by determining the colocalization of 5-HT2a mRNA with enkephalin mRNA or dynorphin mRNA. 5-HT2a mRNA was found in enkephalinergic as well as dynorphinergic neurons. Thus, there does not seem to be a differential distribution of this receptor in the output routes of the ventral striatum. In all of the subregions investigated (core, medial shell, and lateral shell of the nucleus accumbens and the olfactory tubercle), only subpopulations of the total enkephalinergic and dynorphinergic populations were found to contain 5-HT2a mRNA. For enkephalin, the percentage colocalization was higher in the lateral shell (61%) compared with the other subregions (38-45%). For dynorphin, the percentage colocalization was higher in the olfactory tubercle (68%) than in the other subregions (34-43%). The differences in (sub)regional mRNA levels and in colocalization with opioids suggest a considerable regional differentiation in the effects of 5-HT2a-mediated neurotransmission in the striatum. PMID- 9390757 TI - Ultrastructural observations of synaptic connections of vibrissa afferent terminals in cat principal sensory nucleus and morphometry of related synaptic elements. AB - Previous work suggests that slowly adapting (SA) periodontal afferents have different synaptic arrangements in the principal (Vp) and oral trigeminal nuclei and that the synaptic structure associated with transmitter release may be related directly to bouton size. The present study examined the ultrastructures of SA and fast adapting (FA) vibrissa afferents and their associated unlabeled axonal endings in the cat Vp by using intra-axonal labeling with horseradish peroxidase and a morphometric analysis. All SA and FA afferent boutons contained clear, round, synaptic vesicles. All the FA and most SA boutons were presynaptic to dendrites, but a few SA boutons were axosomatic. Both types of bouton were frequently postsynaptic to unlabeled axonal ending(s) containing pleomorphic, synaptic vesicles (P-ending). The size of labeled boutons was larger in FA than SA afferents, but the size of dendrites postsynaptic to labeled boutons was larger for SA than FA afferents. Large-sized FA and SA boutons made synaptic contacts with small-diameter dendrites. The size of FA and SA boutons was larger than that of their associated P-endings. A morphometric analysis made on the pooled data of SA and FA boutons indicated that apposed surface area, active zone number, total active zone area, vesicle number, and mitochondrial volume were highly correlated in a positive linear manner with labeled bouton volume. These relationships were also applicable to unlabeled P-endings, but the range of each parameter was smaller than that of the labeled boutons. These observations provide evidence that the two functionally distinct types of vibrissa afferent manifest unique differences but share certain structural features in the synaptic organization and that the ultrastructural "size principle" proposed by Pierce and Mendell ([1993] J. Neurosci. 13:4748-4763) for Ia-motoneuron synapses is applicable to the somatosensory system. PMID- 9390758 TI - Reevaluation of ipsilateral corticocortical inputs to the orofacial region of the primary motor cortex in the macaque monkey. AB - An anatomical approach to possible areas in the cerebral cortex involved in somatic motor behavior is to analyze the cortical areas containing neurons that connect directly to the primary motor cortex (MI). To define the cortical areas related to orofacial movements, we examined the distribution of cortical neurons that send their axons to the orofacial region of the MI in the macaque monkey. Injections of retrograde tracers into the electrophysiologically identified orofacial region of the MI revealed that labeled neurons were distributed in the following cortical areas: the orbital cortex (area 12), insular cortex, frontoparietal operculum (including the deep part of the cortical masticatory area and the secondary somatosensory cortex), ventral division of the premotor cortex (especially in its lateral part), orofacial region of the supplementary motor area, rostral division of the cingulate motor area (CMA), and CMA on the ventral bank. A number of labeled neurons were also seen in the MI around the injection sites and in the parietal cortex (including the primary somatosensory cortex and area 7b). No labeled neurons were found in the dorsal division of the premotor cortex. Fluorescent retrograde double labeling further revealed virtually no overlap of distribution between cortical neurons projecting to the orofacial and forelimb regions of the MI. Based on the present results, we discuss the functional diversity of the cortical areas related to orofacial motor behavior and the somatotopical organization in the premotor areas of the frontal cortex. PMID- 9390759 TI - Connections of the basal forebrain of the weakly electric fish, Eigenmannia virescens. AB - The organization of the ventral nucleus of the ventral telencephalon (Vv) was examined in the weakly electric fish, Eigenmannia virescens. This nucleus, which is considered the teleost homologue to the basal forebrain nuclei of other vertebrates, was subdivided into dorsal and ventral subdivisions, based upon cytoarchitectonic, immunohistochemical, and connectional criteria. Afferent projections were observed from the medial olfactory bulb as well as the terminal nerve ganglion. Telencephalic afferents to the Vv were very restricted, consisting of the supracommissural and the dorsal intermediate nuclei of the ventral telencephalon, the nucleus taenia, and the medial region of the posterior nucleus of the dorsal telencephalon. However, the major afferents to the Vv were diencephalic, particularly those originating from the rostral preoptic area and other hypothalamic nuclei. Additional afferents included the posterior tubercular nucleus, the locus coeruleus, the medial perilemniscal nucleus, and the periventricular nucleus of the posterior tuberculum. Relatively weak projections were observed from the ventral thalamus and the dorsal posterior thalamic nucleus. As described previously, the diencephalic complex of the central posterior thalamic nucleus/prepacemaker nucleus (CP/PPn), which also has cells that innervate the pacemaker circuitry controlling the production of an electric organ discharge, projects to the Vv. Terminal fields of the Vv were observed to be coextensive with afferent cell groups in the preoptic area, lateral and caudal hypothalamic nuclei, and thalamus. An additional efferent target of the Vv was the pretectal nucleus electrosensorius. That many cell groups that are connected with the Vv are also connected with the CP/PPn, particularly the preoptic and hypothalamic nuclei, suggests that the electrocommunicatory system is intimately linked with basal forebrain limbic pathways. PMID- 9390760 TI - N-methyl-D-aspartate receptor 1 mRNA distribution in the central nervous system of the weakly electric fish Apteronotus leptorhynchus. AB - We have isolated a partial cDNA for the N-methyl-D-aspartate (NMDA) receptor 1 (NMDAR1) subunit from an Apteronotus leptorhynchus brain cDNA library. The A. leptorhynchus cDNA fragment, which corresponds to nucleotides 135-903 within the 5' region of the rat NR1 mRNA, encodes 252 amino acids that are >80% identical to the homologous segments of the rat, human, and duck NR1 proteins. RNAse protection assays revealed that the A. leptorhynchus NR1 mRNA was highly enriched in the forebrain and hypothalamus, with lesser amounts in the brainstem, and very low levels in the cerebellum. In situ hybridization also demonstrated that neurons in the pallial forebrain were highly enriched in NR1 transcripts. High levels of NR1 mRNA were found in pyramidal cells within the optic tectum and octavolateral regions. Pyramidal cells of the electrosensory lateral line lobe had the highest levels of expression, and the NR1 mRNA was found to be selectively enriched in their apical dendrites. PMID- 9390761 TI - Postnatal development and sex difference in neurons containing estrogen receptor alpha immunoreactivity in the preoptic brain, the diencephalon, and the amygdala in the rat. AB - Estrogen has been considered as a key substance that induces sexual differentiation of the brain during fetal and neonatal life in the rat. Thus, to define the brain regions involved in the brain sexual differentiation, we examined the regions where the estrogen receptor (ER) is located in the developing rat brain. We examined immunohistochemical distribution of the cells containing estrogen receptor-alpha (ER-alpha) in the preoptic region, the diencephalon, and the amygdala in male and female rats on postnatal days 1-35 (PD1-PD35). The antibody used recognizes ER-alpha equally well for both occupied and unoccupied forms. ER-alpha immunostaining was restricted to the cell nuclei of specific cell groups. In PD1 rats, ER-alpha-immunoreactive (ER-IR) signals were detected in the lateral septum, the organum vasculosum lamina terminalis, the medial preoptic nucleus (MPN), the median preoptic nucleus, the bed nucleus of the stria terminalis, the hypothalamic periventricular nucleus, the lateral habenula, the posterodorsal part of the medial amygdala nucleus, the posterior part of the cortical amygdala nucleus, the hypothalamic ventromedial nucleus (VMH), the hypothalamic arcuate nucleus, and the posterior hypothalamic periventricular nucleus. The distribution pattern of ER-IR cells in the newborn rat was much the same as that in the adult in the preoptic-hypothalamic and amygdala regions. Moreover, the signals in the MPN and the VMH were stronger in the female than in the male, perhaps reflecting the ability ofestrogen generated by aromatization of testosterone in the male to down-regulate the ER signal. Thus, the brain regions showing sex differences may be sites of sexual differentiation of the brain by aromatizable androgen during the neonatal period. PMID- 9390762 TI - Distribution of N-methyl-D-aspartate and non-N-methyl-D-aspartate glutamate receptor subunits on respiratory motor and premotor neurons in the rat. AB - Glutamate is required for the transmission of inspiratory drive in respiratory premotor and motor neurons. The glutamate receptors (GluRs) responsible for this essential function have yet to be anatomically characterized. We mapped the GluR subtypes expressed by respiratory premotor and motor neurons by using combined immunohistochemistry and retrograde labeling in adult rats. Phrenic motoneurons and bulbospinal ventral respiratory group (VRG) neurons were retrogradely labeled and immunolabeled with subunit-specific antibodies against the N-methyl-D aspartate (NMDA) receptor subtype (NMDAR1) and the non-NMDA receptor subtypes, alpha-amino-3-hydroxy-5-methylisoxazole-4-proprionic acid (AMPA; GluR1, GluR2/3, GluR4) and kainate (GluR5-7). Phrenic motoneurons and bulbospinal VRG neurons showed positive immunolabeling for all five GluR subunits. These results support the hypothesis that NMDA and non-NMDA receptor subtypes underlie the excitation of bulbospinal VRG neurons and phrenic motoneurons. Furthermore, immunolabeling for each receptor subtype demonstrated a unique distribution along the neuronal membrane. Immunoreactivity for AMPA receptor subunits was distributed throughout somata and proximal dendrites, NMDAR1 subunit immunolabeling was localized to somata, and GluR5-7 subunit immunolabeling was confined largely to dendrites. The differential distribution of AMPA, kainate, and NMDA receptors on the somal and dendritic surface of respiratory neurons suggests that the location of glutamatergic synapses along the neuronal surface is an important determinant of glutamate-mediated postsynaptic currents. Consequently, different patterns of glutamatergic excitation of respiratory neurons could be achieved by selective activation of different profiles of GluR subtypes on different portions of the neuronal membrane. PMID- 9390763 TI - Immunocytochemical localization of gamma-aminobutyric acid plasma membrane transporters in the tiger salamander retina. AB - Gamma-aminobutyric acid (GABA) plasma membrane transporters (GATs) play an important role in regulating GABA neurotransmission in the nervous system. The distribution of two GATs, GAT 1 and GAT 3, in salamander retina was investigated by using affinity-purified polyclonal antisera directed to the predicted C terminals of rat GAT 1 and rat GAT 3. GAT 1-immunoreactivity (-IR) was found in type IB and IIB orthotopic bipolar cells (BCs) located in the distal and middle of the inner nuclear layer (INL), respectively; in type IIA and IA amacrine cells (ACs) located in the middle and proximal INL, respectively; and in interplexiform cells and cells in the ganglion cell layer (GCL). No detectable staining was found in horizontal cells (HCs) or in structures resembling Muller cells. GAT 1 immunoreactive fibers were present in the outer plexiform layer (OPL) and inner plexiform layer (IPL) in three bands corresponding to the three bands previously reported to be GABA-IR. GAT 3 antibodies labeled fewer cells and cell types than GAT 1 antibodies. GAT 3-IR was localized to type IIA and IA ACs and cells in the GCL, but not to BCs, HCs, or Muller cell-like structures. There was weak labeling of the OPL and stronger labeling of the IPL, with three distinct bands at the same depth as observed with GAT 1-IR. Double-labeling showed that the majority of GAT 1-IR BCs (88%), ACs (88%), and cells in the GCL (78%) colocalized with GABA IR. The present study provides the first direct evidence of the expression of two GAT subtypes in neurons of nonmammalian retinas. These transporters could regulate GABA neurotransmission by reuptake and termination of GABA's action and, perhaps, by GABA release mechanisms. The presence of GAT 1-IR/GABA-IR bipolar cells further supports our earlier observations that a subgroup of orthotopic bipolar cells are likely to be GABAergic. PMID- 9390764 TI - Identification of gamma-aminobutyric acid-immunoreactive axon endings associated with mesencephalic periodontal afferent terminals and morphometry of the two types of terminals in the cat supratrigeminal nucleus. AB - A previous study has shown that mesencephalic periodontal afferent terminals receive contacts more frequently from axonal endings containing pleomorphic, synaptic vesicles (P-endings) in the supratrigeminal nucleus (Vsup) than in the trigeminal motor nucleus, suggesting that interneurons in Vsup play an important role in modulating the jaw-closing reflex. The present study was attempted to identify neurotransmitters in P-endings associated with mesencephalic periodontal afferents in cat Vsup through the use of intracellular staining of horseradish peroxidase combined with the postembedding immunogold methods. A morphometric analysis was carried out to compare the ultrastructural features of these two types of terminals. Serial sections of 31 labeled boutons and of their associated 38 P-endings were examined. They were processed for postembedding immunogold labeling with antibodies to the neurotransmitter gamma-aminobutyric acid (GABA). The 38 P-endings presynaptic to periodontal afferents showed GABA-like immunoreactivity, but the afferent terminals were free from the labeling. The morphometric analysis indicated that bouton volume, apposed surface area, total active zone size, and mitochondrial volume were smaller in GABA-immunoreactive P endings than in periodontal afferents, but the pooled data of the two types of terminals showed that each synaptic parameter was highly correlated in a positive, linear manner with bouton volume. These observations provide evidence that P-endings presynaptic to mesencephalic periodontal afferents contain the neurotransmitter GABA and that their axoaxonic synapses are organized in accordance with the ultrastructural "size principle" proposed by Pierce and Mendell (Pierce and Mendell [1993] J. Neurosci. 13:4748-4763) on Ia-motoneuron synapses. PMID- 9390765 TI - Distribution of GABAergic premotor nonspiking local interneurones in the terminal abdominal ganglion of the crayfish. AB - The inhibitory neurotransmitter of premotor nonspiking local interneurones in the crayfish terminal abdominal ganglion was investigated physiologically and immunocytochemically. Depolarization of a nonspiking interneurone evoked a hyperpolarization in a uropod motor neurone. The amplitude of hyperpolarization in the motor neurone was gradually decreased under low-calcium/high-magnesium saline. Local pressure injection of gamma-aminobutyric acid (GABA) into the neuropil caused a similar hyperpolarization of the motor neurone. These physiological studies suggested a GABAergic inhibitory interaction between nonspiking interneurones and the motor neurones. Premotor nonspiking interneurones are classified into two subgroups ofposterolateral (PL) and anterolateral (AL) interneurones, and AL interneurones are further divided into three subtypes. A combination of intracellular staining from nonspiking local interneurones with Lucifer yellow and immunocytochemical staining with an antiserum directed against GABA revealed that all the PL interneurones sampled in this study showed GABA-like immunoreactivity. A population of cell bodies (n = 6 11) with a small diameter (15-30 microm) packed together forming a cluster showed GABA-like immunoreactivity, and the cell bodies of most PL interneurones were found in this cluster. To compare the number and the pattern of main branches of PL interneurones, cells were classified into three identifiable sets of interneurones, called PL-1, PL-2, and PL-3. By contrast, about one-half of AL interneurones, especially the third subtype of AL interneurones, which have cell bodies located ventrolaterally in the ganglion, did not show GABA-like immunoreactivity. Furthermore, the position of cell bodies of GABA-immunoreactive AL interneurones was scattered compared to that of PL interneurones. PMID- 9390766 TI - Direct projections from the lumbosacral spinal cord to Barrington's nucleus in the rat: a special reference to micturition reflex. AB - In the present study, direct projections from the lumbosacral cord to Barrington's nucleus in the rat were investigated by using retrograde and anterograde tracing techniques. After injection of cholera toxin B subunit (CTb) into Barrington's nucleus, a number of moderately CTb-labeled neurons were observed in the lumbosacral cord, with a slight ipsilateral dominance; most were located in the spinal parasympathetic and dorsal commissural nuclei of the lumbosacral cord. In addition, some retrogradely labeled neurons were found in the periaqueductal gray (PAG). These findings were confirmed by an anterograde labeling experiment. After biotinylated dextran amine (BDA) was injected into the lumbosacral cord, dense BDA-labeled axon terminals were found in Barrington's nucleus as well as in the PAG. Injection of BDA into the PAG resulted in many BDA labeled terminals in Barrington's nucleus. The present results provided clear evidence for a direct projection from the spinal parasympathetic and dorsal commissural nuclei to Barrington's nucleus that could subserve conveying bladder filling information from the lumbosacral cord to Barrington's nucleus in the micturition reflex of the rat. PMID- 9390767 TI - Starburst cholinergic amacrine cells in the tree shrew retina. AB - In all mammalian retinae studied to date, starburst cholinergic amacrine cells are a consistently occurring cell type. Here, we show that the cone-dominated retina of the tree shrew also has starburst cells with the characteristic radially symmetric branching pattern known from other species. Dendritic field sizes increase from 150 microm in the central retina to 300 microm in the retinal periphery. The characteristic morphology is established early during postnatal development. Labelling the starburst cholinergic cells with an antibody against choline acetyltransferase (ChAT) reveals two dendritic strata in the inner plexiform layer and two corresponding soma populations in the inner nuclear layer (orthotopic) and ganglion cell layer (displaced). These features are present in the adult and in early postnatal stages. In the adult, the density of the orthotopic population as well as the displaced population peaks in the central retina at about 2,200 cells/mm2 and has a peripheral minimum of 400 cells/mm2. These properties are qualitatively similar to those of starburst cells in rod dominated retinae. In contrast to findings in other mammals, we did not see gamma aminobutyric acid (GABA) or glutamic acid decarboxylase 65 kDa (GAD65) immunoreactivity in tree shrew starburst cells. These cells also appear to lack synaptophysin, a ubiquitous synaptic vesicle protein detected in the starburst cells of some other mammals. However, synaptoporin, a homologous synaptic vesicle protein, appears to be present in tree shrew starburst cells. PMID- 9390768 TI - Scanning and transmission electron microscopy of Ruffini endings in the periodontal ligament of rat incisors. AB - The Ruffini organ is an arborized axon ending categorized as a low-threshold stretch receptor. We have previously shown that the lingual periodontal ligament of rat incisors is densely innervated with Ruffini endings. In the present study, fine structures in the surface of the periodontal Ruffini endings and their topographical relationship with the surrounding collagen fibers were observed by a combination of scanning and transmission electron microscopy to analyze the mechanism of the stretch reception. The entire length of the branches of the Ruffini endings, excepting their terminal portions, corresponded well with those depicted by previous investigators in the following points: (1) their cylindrical appearance covered by Schwann cell processes; (2) the presence of numerous axon fingers protruding through gaps in the Schwann sheath and; (3) their isolation from collagen fibers by multilayered basal lamina. On the other hand, tips of the axon branches-together with their Schwann sheaths-became attenuated and projected into tight bundles of collagen, indicating their susceptibility to mechanical deformations of the surrounding tissue. Margins of the axon terminals were conspicuously ruffled with long tongue-like projections of Schwann cells. The Schwann cell tongues twined around collagen bundles in their distal portions, and associated closely with fine axon projections in their proximal portions, suggesting their involvement in the mechanical transmission of stimuli to axon terminals. PMID- 9390769 TI - Sensitive period for lesion-induced reorganization of intracortical projections within the vibrissae representation of rat's primary somatosensory cortex. AB - Previous experiments from this laboratory demonstrated that intracortical connections in lamina IV of the rat primary somatosensory cortex (SI) are most dense outside the patches of cytochrome oxidase (CO) staining that correspond to the mystacial vibrissae. This pattern of intracortical connections becomes apparent on postnatal day 4 (P-4), at least 2 days after the appearance of the vibrissae-related pattern of thalamocortical afferents. Transection of the infraorbital nerve (ION) on the day of birth (P-0) disrupts both the CO and intracortical projection patterns. This series of experiments was undertaken to determine whether the patterning of either thalamocortical afferents or intracortical projections defines the end of the period over which peripheral damage can alter intracortical projections in lamina IV of SI. The infraorbital nerve (ION) was transected in different cohorts of rats on P-1 through P-5, and animals were allowed to survive > or =45 days, at which time biotinylated dextran amine (BDA) injections were made into the SI. After 7 days, animals were killed, and alternate cortical sections were processed for the demonstration of BDA or CO. Transection of the ION on P-1 or P-2 altered the patterning of both CO and intracortical connections in the SI. In contrast, cutting the ION on P-3 left the pattern of CO densities in the SI intact, but significantly altered the patterning of intracortical connections. Transection of the nerve on P-5 resulted in qualitatively and quantitatively normal patterns of both CO densities and BDA labelled intracortical projections. These results indicate that the establishment of a stable barrel pattern in layer IV of the SI is not sufficient for normal adult patterning of intracortical projections in this lamina. However, once the mature pattern of intracortical projections in layer IV is established, ION lesions can no longer alter it. PMID- 9390770 TI - Elevated 4-hydroxynonenal in ventricular fluid in Alzheimer's disease. AB - 4-Hydroxynonenal (4-HNE), an aldehyde by-product of the peroxidation of fatty acids, has been shown to have toxic properties for neurons in culture. In light of increasing evidence that oxidative stress contributes to the neurodegenerative process in Alzheimer's disease (AD), we quantified levels of free and protein bound 4-HNE in the ventricular fluid from 19 AD subjects and 13 control subjects by high-pressure liquid chromatography and dot-blot immunoassay. Free 4-HNE levels were found to be significantly elevated in the ventricular fluid of AD subjects compared with control subjects (p = 0.0096). These results demonstrate increased lipid peroxidation in AD brain and suggest a role for 4-HNE in the neurodegenerative process. PMID- 9390771 TI - Alzheimer's disease: in vivo detection of differential vulnerability of brain regions. AB - The severe cognitive impairment during the later stages of Alzheimer's disease is usually preceded by a selective disturbance in the ability to remember new experiences. With quantitative, high-resolution magnetic resonance imaging techniques, it is now possible to determine, in vivo, differences in the pattern of anatomical changes that might reflect behavioral symptomatology during different stages of the disease. In the present investigation, magnetic resonance imaging examinations were carried out in aged controls and in clinically diagnosed Alzheimer's disease patients who were divided into three groups based upon dementia severity. Atrophy of the hippocampal formation, a region important for memory function, was observed even in Alzheimer's disease patients with the mildest dementia. With more prominent dementia, atrophy extended to the parahippocampal gyrus and the temporal neocortex. PMID- 9390772 TI - GLUT-1 expression in the cerebra of patients with Alzheimer's disease. AB - To investigate the molecular basis of reduced GLUT-1 concentration of the blood brain barrier in patients with Alzheimer's disease (AD), the GLUT-1 mass, mRNA content, and structure were studied in eight patients with AD and seven age matched controls. The results indicate that the 55-kDa GLUT-1 is significantly reduced in AD without a significant change in GLUT-1 mRNA concentrations. Because in some animal models changes in GLUT-1 expression is associated with changes in GLUT-1 mRNA structure, the length of the poly(A) tail of the GLUT-1 mRNA was estimated with a reverse transcription-polymerase chain reaction technique. The length of poly(A) tail of GLUT-1 mRNA in AD subjects was not significantly different from the controls. It is concluded that the AD-related change in GLUT-1 expression is not the result of altered poly(A) length of GLUT-1 mRNA. PMID- 9390773 TI - Low initial tau phosphorylation in human brain biopsy samples. AB - A rapid reversible tau phosphorylation at Ser 396/404 was observed in adult human cortical biopsy tissue and rat primary cortical cell cultures. Tau phosphorylation increased usually during the first 20-30 min in phosphate buffered saline, followed by a decrease. The time course of tau phosphorylation and dephosphorylation in biopsy tissue could be lengthened by culturing in defined, oxygenated medium, instead of in phosphate-buffered saline. Phosphorylation of total protein in biopsy tissue occurred in two phases, with peaks at 30 and 90 min. The first peak of total protein phosphorylation coincided with the peak of tau phosphorylation, although both the first and second peaks of total protein phosphorylation coincided with the first and second peaks of neurofilament-H phosphorylation. PMID- 9390774 TI - Age diminishes performance on an antisaccade eye movement task. AB - Tthe antisaccade eye movement task, which has been linked to frontal lobe function, presents a target in one visual field and asks subjects to move their eyes to the same location in the opposite field. The task requires inhibition of the reflexive prosaccade to the cue, initiation of the antisaccade to the opposite field, and visuo-spatial memory of the cue location. Forty-two subjects from 19-79 years of age performed this task and a control task, visually guided saccades to the cue itself, to determine which functions are affected by aging. The time to initiate antisaccades increased linearly with age at a rate greater than the time to initiate visually guided saccades. This difference suggests that the processing time to inhibit the incorrect movement to the cue is selectively increased with age. Older subjects also made more incorrect prosaccadic movements to the cue, a finding consistent with the loss of inhibitory processing capacity. The accuracy of movements did not change, which suggests that visuo-spatial memory is unaffected by aging. PMID- 9390775 TI - Reduced efficacy of growth hormone-releasing hormone in modulating sleep endocrine activity in the elderly. AB - In aging, a decline in sleep continuity, a decreased slow wave sleep, an earlier nocturnal cortisol rise, and a blunted growth hormone (GH) secretion occur. Pulsatile administration of GH-releasing hormone (GHRH) in young controls enhanced slow wave sleep and suppressed cortisol release. We administered GHRH 4 x 50 microg or placebo i.v. to 13 healthy seniors (5 women, 8 men, mean age 69.3 y +/- 8.3 SD). We observed significantly reduced nocturnal awakenings and an increased first non-rapid-eye-movement sleep period. In a subgroup (n = 9), we found a significant activation of GH secretion but unchanged cortisol secretion. Our data underscore that GHRH is capable of promoting sleep in the elderly, but much less than in young subjects. Contrasting to young subjects, the hypothalamic pituitary-adrenocortical system remains unaffected by GHRH in the elderly. These results provide further evidence that a decrease in the efficacy of GHRH is involved in the biological mechanisms underlying aging. PMID- 9390776 TI - Biphasic and region-specific MAO-B response to aging in normal human brain. AB - Variations of monoamine oxidases (MAO) A and B were studied during aging in 27 human subjects (age range 17-93 years) in 18 brain structures of temporal cortex, frontal gyrus, hippocampal formation, striatum, cerebellum, and brainstem. [3H]Ro41-1049 and [3H]lazabemide were used as selective radioligands to image and quantify MAO-A and MAO-B respectively by enzyme autoradiography. Postmortem delay or time of tissue storage did not affect MAO-A or MAO-B levels. There was, moreover, no evidence of sexual dimorphism. A marked age-related increase in MAO B was observed in most structures. This increase started at the age of 50-60 years. Before this age, MAO-B levels were constant in all structures studied. MAO B-rich senile plaques were observed in some cortical areas but they did not significantly influence the age-related MAO-B increase. Surprisingly, no age related MAO-B changes were observed in the substantia nigra. In contrast to MAO B, no clear age-related changes in MAO-A were observed, indicating an independent regulation of the two isoenzymes, also suggested by the cross-correlation analysis of these data. PMID- 9390777 TI - Aging, dominance history, and social behavior in Java-monkeys (Macaca fascicularis). AB - The aim of this study was to investigate the influence of the dominance history of socially housed Java-monkeys on the aging process. In monkeys, social subordinance is generally associated with elevated levels of cortisol, which, in turn, have been suggested to influence cognitive decline. As cognitive skills are necessary for successful social life, we investigated the effect of old age in relation to the dominance history of the animals on their social behavior by comparing old females with their younger daughters. Old age, especially in combination with a history of low rank, led to a withdrawal from social interactions with unfamiliar animals and to a decrease in amounts of aggression received. Still, however, old animals showed an increase in behaviors associated with arousal. A reduced ability to deal with complex social interactions, caused by a decline in information processing abilities, is suggested as an explanation for these results. PMID- 9390778 TI - T2-weighted MRI studies of mouse lemurs: a primate model of brain aging. AB - Previous histological and behavioral studies of aging mouse lemurs have demonstrated changes similar to those observed in elderly humans and in patients with Alzheimer's disease. We explored 18 animals of ages 6 months to 9 years. Axial T2-weighted images of the brain were performed on a 4.7 Tesla Bruker Biospec 47/30 system. We estimated cerebral atrophy by adding measures of high signal areas characteristic of cerebrospinal fluid (interlobular and sylvian fissures, lateral and third ventricles) of four contiguous cortical slices. We observed a significant increase of cerebral atrophy with aging and one case of an apathetic 8-year-old animal presenting a considerably higher cerebral atrophy. We also observed high correlations between decreased signal intensities and age for the pallidum, the substantia nigra, and the putamen. These results suggest that aging mouse lemurs present similar magnetic resonance images of cerebral alterations to those encountered in aging humans and that high-field T2-weighted magnetic resonance images can help in the early detection, in vivo, of animals suspected of pathological aging. PMID- 9390779 TI - Life-long dietary restriction attenuates age-related increases in hippocampal glial fibrillary acidic protein mRNA. AB - The expression of astrocyte-specific glial fibrillary acidic protein increases after experimental lesions and is elevated throughout the brain in aged rodents and primates. Clusterin (ApoJ) expression increases in astrocytes and microglia after lesions, but changes during aging have not been reported. Dietary restriction (DR) delays the onset and progression of many age-related physiological deficits in rodents. This study showed that the age-related increase in glial fibrillary acidic protein mRNA in the hippocampus was attenuated in 24-month-old male Fischer 344 rats subjected to a 50% DR beginning at 6 weeks of age. ApoJ mRNA expression in astrocytes was unchanged by DR. These results demonstrate that DR can delay neurodegeneration in aged rats as assessed by a marker of reactive astrogliosis. PMID- 9390780 TI - Antidepressants restore hypothalamic-pituitary-adrenal feedback function in aged, cognitively-impaired rats. AB - Aged, cognitively-impaired rats (and humans) show hypothalamic-pituitary-adrenal (HPA) hyperactivity that correlates with hippocampal damage. The resultant increase in plasma glucocorticoid exposure is thought to contribute to impaired hippocampal function and to potentiate hippocampal neuron death. In young, adult rats antidepressant drugs increase corticosteroid receptor expression in brain regions known to regulate the HPA axis, leading to increased negative-feedback control and decreased HPA activity. In the present study we examined basal levels of plasma adrenocorticotropin hormone (ACTH) and corticosterone in aged, cognitively-impaired (AI), aged, cognitively-unimpaired (AU) and young, adult (Yg) rats. Plasma ACTH and corticosterone levels were significantly elevated in the AI rats, but only in samples obtained during the diurnal peak. Five weeks of treatment with desipramine (15 mg/kg) significantly reduced evening levels of both ACTH and corticosterone in all groups, and eliminated the group differences. We then examined delayed, glucocorticoid negative feedback in these animals. Among vehicle-treated animals, a bolus injection of corticosterone (10 mg/kg), administered 3 hours prior to testing, completely inhibited the plasma ACTH response to restraint in AU and Yg, but not AI animals. In contrast, plasma ACTH responses to restraint were completely inhibited in AI rats following chronic treatment with desipramine. These findings indicate that the antidepressant, desipramine, decreases HPA activity and increases glucocorticoid negative feedback sensitivity in AI rats, suggesting that antidepressant drugs may form a useful therapeutic approach to HPA dysfunction in elderly human populations. PMID- 9390782 TI - Titers of murine leukemia virus are higher in brains of SAMP8 than SAMR1 mice. AB - To elucidate possible causes of premature aging seen in a strain of senescence accelerated prone (SAMP8) mice, levels of murine leukemia virus (MuLV) were quantitated in various tissues of SAMP8 by an SC-1/UV plaque assay. MuLV levels in SAMP8 tissues were compared to those seen in the closely related SAMR1 strain, which is resistant to premature aging. MuLV titers were found to be higher in blood and spleen and much higher in brain of SAMP8 than in the same tissues of SAMR1. MuLV levels were seen to increase in SAMP8 brain with increasing age. Virus typing experiments indicated that the MuLV from SAMP8 brain is N-tropic, as is the MuLV seen in the AKR strain, one of the SAM progenitor strains. MuLV from SAMP8 brain was able to grow well in SAMR1 mouse embryo cells, indicating that it may be possible to infect SAMR1 mice with the SAMP8 MuLV to determine the effects of the virus on aging of SAMR1 mice. PMID- 9390781 TI - NMDA receptor activation in the aged rat: electrophysiological investigations in the CA1 area of the hippocampal slice ex vivo. AB - The effects of aging on activation of N-methyl-D-aspartate (NMDA) receptors were studied in the CA1 field of hippocampal slices from young (2-4 months old) and aged (25-32 months old) Sprague-Dawley rats with the use of ex vivo extra- and intracellular electrophysiological recording techniques. No significant age related changes of the unitary NMDA-receptor mediated excitatory postsynaptic potentials (EPSPs), recorded from the pyramidal cells after stimulation of the stratum radiatum in a magnesium-free medium and isolated in the presence of the non-NMDA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione, were found. Simultaneously, the magnitude of synaptic plasticity which involved NMDA receptor activation was not altered. No significant age-related modifications in the mechanisms controlling glutamate release and of postsynaptic NMDA receptor responsiveness were revealed. Considering the 30-40% decrease in NMDA binding sites in the aged hippocampus, our results suggest the occurrence of compensatory mechanisms which are discussed. PMID- 9390783 TI - Preserved number of entorhinal cortex layer II neurons in aged macaque monkeys. AB - The perforant path, which consists of the projection from the layer II neurons of the entorhinal cortex to the outer molecular layer of the dentate gyrus, is a critical circuit involved in learning and memory formation. Accordingly, disturbances in this circuit may contribute to age-related cognitive deficits. In a previous study, we demonstrated a decrease in N-methyl-D-aspartate receptor subunit 1 immunofluorescence intensity in the outer molecular layer of aged macaque monkeys. In this study, we used the optical fractionator, a stereological method, to determine if a loss of layer II neurons occurred in the same animals in which the N-methyl-D-aspartate receptor subunit 1 alteration was observed. Our results revealed no significant differences in the number of layer II neurons between juvenile, young adult, and aged macaque monkeys. These results suggest that the circuit-specific decrease in N-methyl-D-aspartate receptor subunit 1 reported previously occurs in the absence of structural compromise of the perforant path, and thus may be linked to an age-related change in the physiological properties of this circuit. PMID- 9390784 TI - Cerebrospinal fluid C3a increases with age, but does not increase further in Alzheimer's disease. AB - Complement activation is present in the brain in Alzheimer's disease (AD), and C1q concentrations are decreased in AD cerebrospinal fluid (CSF). To determine whether concentrations of other complement proteins are also altered in AD CSF, we measured concentrations of C3a and SC5b-9 in CSF from patients with probable AD (n = 19), normal aged controls (n = 11), and normal younger controls (n = 15). C3a concentrations were similar between AD and aged controls, but threefold higher than in younger controls (p < 0.05 vs. both groups). A similar pattern was found with SC5b-9, though the increase was only twofold and statistically significant only for AD vs. younger controls. These results suggest that an increased generation of complement proteins in localized areas of the AD brain does not result in elevated concentrations of these proteins in CSF, compared with age-matched controls. Increased C3a (and, to a lesser extent, SC5b-9) in aged controls may be due to increased complement activation, increased central nervous system production, and/or blood-brain barrier leakage of these proteins. PMID- 9390786 TI - Nonlinearity of mechanoelectrical transduction of outer hair cells as the source of nonlinear basilar-membrane motion and loudness recruitment. AB - The sound-induced travelling wave in the mammalian cochlea is believed to be enhanced and sharpened by a positive-feedback mechanism, causing the passive, linear growth function of the basilar membrane (BM) to become nonlinear. Based on direct measurements of the receptor potential of isolated outer hair cells, it is shown here how nonlinear BM motion might be due predominantly to the nonlinear growth function of the receptor potential. Since intensity coding in the inner ear is supposed to depend on an interaction of nonlinear BM motion with afferent fibres of different synaptic thresholds, intensity coding is expected to be directly dependent on the mechanoelectrical transduction of outer hair cells (OHC). According to the present experimental data and the feedback concept of outer hair cell action, disruption of the mechanoelectrical transduction of OHC leads to both a reduction of gain and linearizing of the response; that is, to both hearing loss and loudness recruitment. PMID- 9390785 TI - Intracellular inositol (1,4,5)-trisphosphate receptor levels are preserved in Alzheimer's disease platelets. AB - An increasing number of signal transduction disturbances have been reported in Alzheimer's disease. These changes are not restricted to histopathologically changed brain areas but are seen also in peripheral tissues. One of the most severe disturbances is a loss of calcium-mobilizing intracellular inositol(1,4,5) trisphosphate receptors in Alzheimer cerebellar and cortical tissues. In the present study, the binding of [3H]inositol(1,4,5)trisphosphate ([3H]Ins(1,4,5)P3) to the calcium-mobilizing inositol(1,4,5)trisphosphate receptors in platelet membranes from eight Alzheimer's disease patients and eight control subjects were investigated to determine its possible role as a biological marker in Alzheimer's disease. It was found that there were no significant difference in [3H]Ins(1,4,5)P3 binding with respect to the number of sites measured at different protein concentrations or to the sensitivity of the binding to inhibition by nonradioactive Ins(1,4,5)P3 between Alzheimer disease platelets and controls. It is concluded that inositol(1,4,5)trisphosphate receptor levels are preserved in platelets from patients with Alzheimer's disease. PMID- 9390787 TI - The temporal window of two-tone facilitation in onset units of the ventral cochlear nucleus. AB - Effects of a tone, not at the best frequency (BF), on the responses of onset units to BF tones were recorded in the ventral cochlear nucleus of the guinea pig. The off-BF tone was at a fixed non-excitatory sound level. When the two tones were gated simultaneously, a marked threshold facilitation was observed; however, no facilitation was observed if the off-BF tone burst was delayed by 5 10 ms. Facilitation in some units declined and was either absent or only just detectable, when the start of the off-BF tone preceded that of the BF by more than 10 ms. However, the responses of the majority of onset units were facilitated when the off-BF tone preceded the BF tone by as much as 20 ms. Similar results were obtained when the roles of the BF and off-BF tones were reversed. These data suggest that depolarization due to BF inputs is of shorter duration than that due to off-BF inputs. PMID- 9390788 TI - Immunologically defined component of the circumferential ring around the cuticular plate in mammalian hair cells. AB - A monoclonal antibody (CAR) was raised by in vitro immunisation to a component of the circumferential actin ring that is associated with the apical junctions encircling the cuticular plates of mammalian hair cells. On western blots it bound a protein band at about 42 kD, equivalent to the normal location of actin, but it did not label the paracrystalline bundle of actin filaments in the stereocilia, the complex actin filament gel that forms the cuticular plate or the filamentous actin in the cell cortex. When applied to whole mounts of the auditory sensory epithelium in the guinea pig it provided a clear, unambiguous map of the distribution of inner and outer hair cells. In this respect, it can provide an accurate guide to patterns of hair cell differentiation and repair. CAR cross-reacted with the membrane-associated cytoskeleton in selected cells from a wide range of other tissues. PMID- 9390789 TI - Localization of the hair-cell-specific protein fimbrin during regeneration in the chicken cochlea. AB - Fimbrin is an actin-binding protein that organizes actin filaments into parallel bundles. We have used fluorescence immunocytochemistry and confocal laser scanning microscopy to demonstrate that the detection of fimbrin in the chicken basilar papilla (BP) by this method was limited to the hair cells. These experiments indicate that it was concentrated in the bundles of stereocilia. In addition, there was no evidence for the presence of fimbrin in the supporting cells, border cells, or hyaline cells. During recovery of the BP following acoustic trauma, the appearance of fimbrin in regenerating hair cells was first seen approximately 96 h after the onset of the sound exposure. This was the time at which new hair cells first appear in the BP. Reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect transcription of the fimbrin gene in both control and sound-damaged BP. Semiquantitative RT-PCR indicated no detectable changes in fimbrin transcription levels 2 or 5 days after the onset of pure-tone exposure. These data indicate that fimbrin can be used immunocytochemically as a hair-cell-specific marker during regeneration in the chicken cochlea. PMID- 9390790 TI - Localization of dopamine-beta-hydroxylase-like immunoreactivity in the superior olivary complex of the rat. AB - The noradrenergic (NA) innervation of the superior olivary complex (SOC) was investigated using a monoclonal antibody against dopamine-beta-hydroxylase (D beta H), the synthesizing enzyme for noradrenaline. In addition, positive labelling of D beta H in the inferior colliculus, the lateral lemniscus (LL) and the cochlear nucleus were documented. The antibody was detected using the highly specific and sensitive ABC immunocytochemical method. The specific aim of the study was to show the distribution of putative NA cells and fibres in the SOC in greater detail than had been previously reported. All investigated auditory regions contained D beta H-positive fibres in varying densities. All superior olivary and periolivary nuclei showed NA innervation by mainly fine, weakly staining fibres. Both nuclei of the LL were also found to contain D beta H positive fibres. D beta H-positive cell bodies were found in the region of the ventral lateral lemniscus and immediately lateral and dorsal to the lateral superior olivary nucleus. Thus the NA innervation of the superior olivary complex could originate from these regions and/or the locus coeruleus. PMID- 9390791 TI - Tectorial membrane repair in the quail following multiple exposures to intense sound. AB - Adult quail were exposed one, two or three times to an intense octave band noise. Birds were sacrificed 0 or 14 days after the last exposure and their ears processed for scanning electron microscopy (SEM). SEM observations demonstrated a patch lesion in every ear at 0 and 14 days after exposure. In the single-exposed ear immediately after noise, the lesion lacked any tectorial membrane (TM). At 14 days of recovery, the site of injury was covered by a partially regenerated TM (referred to as the honeycomb). However, the patterns of TM damage and repair varied among the twice- and thrice-exposed papillae. At 0 days after exposure, the multiple-exposed ear displayed a lesion with two segments. In one portion the TM was missing, and in the other portion the TM was present as one or two honeycomb layers. Fourteen days later, a new honeycomb was observed underneath the remaining honeycomb. These results suggest that the quail is able to repeatedly repair its TM following multiple noise exposures. PMID- 9390792 TI - Activation of aminoglycoside antibiotics to cytotoxins. AB - We have previously postulated an enzymatic transformation of gentamicin (to a metabolite or an 'activated' molecule) as part of its ototoxic action. Here we test with eight aminoglycosides whether the proposed mechanism applies to these antibiotics as a group. Drugs were activated by incubation with a subcellular fraction from liver, and cytotoxicity was tested in a bioassay using isolated outer hair cells from guinea pig. None of the aminoglycosides compromised the viability of the cells when assayed directly, i.e. without a preceding activation. In contrast, all clinically ototoxic aminoglycosides tested were significantly cytotoxic following the incubation. Neamine, considered to be non ototoxic, did not yield a cytotoxin. A subcellular fraction from cochlear lateral wall tissues also converted gentamicin to a cytotoxin. The results support the hypothesis that activation of aminoglycosides precedes their toxic actions and demonstrate that the capability for activation is not confined to liver. PMID- 9390793 TI - Incomplete recovery of chicken distortion product otoacoustic emissions following acoustic overstimulation. AB - Distortion product otoacoustic emissions (DPOAEs) were measured in chickens before and after exposure to a 525-Hz pure tone (120 dB SPL, 48 h). The exposure caused extensive hair cell loss and destroyed the tectorial membrane along the abneural edge of the basilar papilla in the low-to-mid-frequency region of the cochlea. Although the lesion was restricted, DPOAEs were greatly depressed at all frequencies immediately after the exposure. The high-frequency DPOAEs gradually recovered to preexposure values after the exposure; however, there was little or no improvement in DPOAEs at test frequencies equal to or slightly above the exposure frequency even after 16 weeks of recovery. By 28 days of recovery, the previously damaged region of the basilar papilla had been repopulated by hair cells and the lower honeycomb layer of the tectorial membrane had regenerated, but not the upper fibrous layer. The upper fibrous layer of the tectorial membrane was still missing after 16 weeks of recovery and the region of damage corresponded closely to the frequency regions where the DPOAEs were depressed. PMID- 9390794 TI - Binaural unmasking returns to normal in teenagers who had otitis media in infancy. AB - Several studies have shown that the binaural masking level difference (MLD) is reduced in children who have a history of recurrent otitis media (OM) in infancy. In this prospective study we have retested the MLD in two groups of teenagers who were originally tested 6 years ago. Members of one of these groups (OM group; n = 26) had at least 5 recorded episodes of OM before the age of 5 years. The other group (controls; n = 17) had no known history of OM. The mean MLD (500-Hz tone in a narrow band noise masker) of the OM group was, at 6-12 years of age, significantly lower than that of the controls. In this study (at ages 12-18 years), we found no difference between the mean MLDs of the OM and control groups. These results show that the reduced MLD that occurs following OM in infancy can recover in later childhood. PMID- 9390795 TI - On the clinical relevance of mismatch negativity: results from subjects with normal hearing and cochlear implant users. AB - Mismatch negativity (MMN) provides an objective measure for evaluating subjects with problems related to speech processing. For a valid neurophysiological profile of speech-processing mechanisms, an efficient procedure to elicit MMNs is needed. In Experiment 1 of this study, MMN recordings were conducted in adults with normal hearing on the effects of decreasing the duration of the interstimulus interval (ISI). Shortening ISI duration does not seem to have a high impact on the individual MMN quality, whereas it does influence group MMN quality. In Experiment 2, MMNs were elicited in a group of cochlear implant users by using a speech sound contrast/ba/-/da/. A group of good performers produced a significant MMN, whereas a group of moderate performers did not. There seems to be a relation between speech perception ability and MMN quality. To fundamentally understand the effects of electrical stimulation of the inner ear and to clinically adjust rehabilitation, diverse data are needed on different aspects of auditory processing. Optimizing the procedure to elicit and MMN is therefore of great clinical value. PMID- 9390796 TI - Diagnosis of endolymphatic hydrops by low-frequency masking. AB - Low-frequency masking is a new method for the diagnosis of endolymphatic hydrops. A short acoustic stimulus and a low-frequency masker tone are applied to the same ear in an adjustable phase relationship. We recorded phase-dependent masked thresholds from normal-hearing subjects, and patients with Meniere's disease and sensory hearing loss without vertigo. In normal hearing, there is a mean maximal difference in masking (modulation depth) of 28 dB between the phase delays of 0 degree and 270 degrees. In patients with sensory hearing loss without vertigo, modulation depth is reduced due to recruitment. In Meniere cases, the phase dependence may be totally absent and varies as the disease progresses. Therefore, repeated measurements of masking are required: patients and subjects with normal hearing were tested for a period of 1 year. Also, modulation depth is significantly reduced in the contralateral nonsymptomatic ears of Meniere patients. The results indicate that low-frequency masking is a quick, noninvasive and relevant method for the diagnosis of endolymphatic hydrops. PMID- 9390797 TI - The importance of thyroid hormone for auditory development in the fetus and neonate. AB - It seems that many auditory maturational events are regulated by thyroid hormone since elevation in thyroid hormone level always precedes the onset of hearing in the fetus-neonate; low thyroid activity in the developing human fetus or rat neonate leads to hearing loss; earlier, elevated thyroid levels in rat neonate lead to earlier onset of hearing. The hormone, bound to its receptors in the nucleus, acts as a transcription factor activating genes which lead to the synthesis of several proteins and enzymes involved in the structural and functional development of many tissues (e.g. brain, heart, kidney, skeletal muscle) including the ear. Several types of congenital hearing loss of unexplained etiology may be due to abnormalities in one or more stages of this gene cascade since several types of congenital hearing loss have been shown to involve defects in genes related to these events. PMID- 9390798 TI - Noise-induced cochlear hypoxia is intensity dependent, correlates with hearing loss and precedes reduction of cochlear blood flow. AB - Anesthetized and artificially ventilated guinea pigs were exposed to broad-band noise of 95, 101, 106 or 115 dB SPL for 30 min and studied for 180 min after cessation of noise. The partial pressure of oxygen (pO2) in the perilymph, the cochlear blood flow (CoBF) and auditory-evoked potentials were continuously recorded. Arterial blood pressure, electrocardiogram, inspiratory and expiratory gas levels, arterial blood gas levels and acid-base status were kept stable to exclude influences of these parameters on cochlear parameters. Exposure to 95 dB SPL did not affect perilymphatic pO2 or CoBF. Cochlear microphonics (CMs) were reduced, but compound action potentials of the auditory nerve (CAPs) and auditory brainstem potentials (ABRs) increased after exposure to this low-level noise. Perilymphatic pO2 decreased during exposure to 101 dB SPL and then further decreased during the subsequent 60 min after cessation of the noise. CoBF did not change significantly during and 30 min after noise but then paralleled the decline of perilymphatic pO2. However, both parameters showed a clear indication of recovery in the second and third hours after noise. At 101 dB SPL, CMs were again reduced immediately, CAPs were unaltered and ABRs again increased. Exposure to 106 and to 115 dB SPL resulted in a decrease in both perilymphatic pO2 and CoBF; this decrease began during the exposure but became progressively worse after the noise. Hearing loss was observed immediately with exposure and showed no signs of further deterioration after cessation. The observed time courses of changes are important. They reveal that hearing loss and cochlear hypoxia precede reduction in CoBF due to noise exposure. The potential mechanisms underlying these effects are discussed. PMID- 9390799 TI - Neurophysiological correlate of the auditory after-image ('Zwicker tone'). AB - The 'Zwicker tone' (ZT) is an auditory after-image that can be evoked most effectively when a band-suppressed noise (relative width of gap 1/3 octave) presented for a certain period of time has been switched off. The sensation of this purely monaural phenomenon is that of a pure tone with a frequency corresponding to the center frequency of the gap and an equivalent level of 10-15 dB above auditory threshold. The sensation decays gradually; it may last as long as 10 s depending on how long the evoking noise was presented. The search for a physiological correlate has been futile so far, probably because the search was confined to more peripheral levels of the auditory system (inferior colliculus). A neuromagnetic study was performed in normal-hearing subjects in order to look for a neurophysiological correlate of the ZT in the auditory cortex. With a stimulation paradigm especially designed for this study, we have been able to isolate poststimulus activity which appears to be related to the ZT and which originates in the supratemporal auditory cortex. It is a sustained neuromagnetic activity that shows a clear-cut dipolar field distribution, and it appears that this activity has certain similarities with the tone-evoked auditory sustained response. The hypothesis is put forward that during the sensation of the ZT a process takes place in the auditory cortex which is similar to that underlying the sustained response, and which gives rise to the sensation of the ZT. In contrast to the sustained response, however, which is due to neural activity evoked by an external acoustic stimulus, the sustained activity associated with the ZT is due to a temporary absolute or relative reduction of neural activity originating from those regions in which the ZT exciting stimulus caused an adaptation. These differences in neural activity cannot be distinguished by the auditory system from a corresponding external acoustic signal. Preliminary studies in patients suffering from tonal tinnitus yielded results which exhibit a certain similarity with those obtained in the ZT experiment. PMID- 9390800 TI - Measures of binaural hearing in children with a history of asymmetric otitis media with effusion. AB - This study addresses the effect of early asymmetric hearing loss in children, owing to otitis media with effusion (OME), on binaural hearing. Five children who had suffered from predominantly unilateral OME between the ages of 2 and 4 years and who were not treated for OME at any time participated in this study when they were about 12 years of age. All children had normal hearing at the time of testing. Data were compared to normative values obtained previously from normal hearing adults. We measured the auditory brainstem response (ABR), the binaural interaction component (BIC) in the ABR, the masking level difference (MLD) and the suppression of transient evoked otoacoustic emissions (OAEs) with contralateral noise stimulation. The results indicated that the children's ABRs and BICs were comparable to normative data, that there was evident suppression of transient evoked OAEs in 4 of the 5 children and that the children's MLD values were within the normal (adult) range. The present results therefore do not support the presence of long-term auditory processing deficits induced by early asymmetric OME in man. PMID- 9390801 TI - Cervical vertigo--reality or fiction? AB - Neck afferents not only assist the coordination of eye, head, and body, but they also affect spatial orientation and control of posture. This implies that stimulation of, or lesions in, these structures can produce cervical vertigo. In fact, unilateral local anesthesia of the upper dorsal cervical roots induces ataxia and nystagmus in animals, and ataxia without nystagmus in humans. If cervical vertigo exists outside these experimental conditions, it is obviously characterized by ataxia and unsteadiness of gait, and not by a clear rotational or linear vertigo. Neurological, vestibular, and psychosomatic disorders must first be excluded before the dizziness and unsteadiness in cervical pain syndromes can be attributed to a cervical origin. To date, however, the syndrome remains only a theoretical possibility awaiting a reliable clinical test to demonstrate its independent existence. PMID- 9390802 TI - 2f1-f2 distortion product otoacoustic emissions in White Leghorn chickens (Gallus domesticus): effects of frequency ratio and relative level. AB - The effects of primary tone frequency ratio (f2/f1 ratio) and relative level (L2/L1) on the amplitude of the cubic difference tone (CDT: 2f1-f2) distortion product otoacoustic emissions (DPOAEs) were investigated in adult White Leghorn chickens (Gallus domesticus). In experiment 1, 9 f2/f1 ratios ranging from 1.05 to 1.8 were investigated. Measurements were obtained from both ears of 4 chickens at 7 f1 frequencies ranging from 0.8 to 4.0 kHz. The primary tones were equal in level, and varied from 20 to 80 dB SPL. The mean CDT amplitude increased with increasing primary tone level once the measurement noise floor was exceeded. The input/ output functions assumed one of two shapes: one in which there was a systematic increase in DPOAE amplitude with increasing primary tone level, and the other in which there was a plateau in the input/output function near 65-70 dB SPL. At the highest primary tone level (80 dB SPL), there was a decrease in the CDT amplitude with increasing f2/f1 ratio. At high primary tone levels, the f2/f1 ratio which produced the largest CDT was 1.05 or 1.1, while at lower primary tone levels the largest CDT occurred at f2/f1 ratios of 1.2-1.3. In experiment 2, L2 was held constant at 70 dB SPL, and L1 varied from 50 to 80 dB SPL. For f1 frequencies of 0.8 and 3.2 kHz, there was an increase in the CDT amplitude with increasing L1, followed by an asymptote at higher levels. In contrast, for 1.6 and 2.0 kHz f1 frequencies, the amplitude increased, plateaued and then increased again at higher levels. Informal measurements suggest that spontaneous otoacoustic emissions (SOAEs) are rarely seen in chickens. However, a reliable SOAE was observed in 1 chicken, which could be suppressed by external sounds and anoxia. PMID- 9390803 TI - A method for determining interaural attenuation in animal models of asymmetric hearing loss. AB - Asymmetric or unilateral sensorineural hearing loss is an important hall-mark of various forms of sensorineural hearing loss. Animal research regarding the etiology and mechanism of these disorders often requires hearing estimates in each ear of experimental animals. Monaural auditory testing of animals with experimentally induced unilateral hearing loss therefore requires prior knowledge of interaural attenuation (IAA) to facilitate contralateral masking. The purpose of this study is to describe a method of determining frequency-specific IAA data and to present relevant information obtained in the rats--a frequently used animal in studies of acquired sensorineural hearing loss. A custom-made sound source was designed to accomplish threshold determination at important frequencies in the dynamic range of rats. Six male Long-Evans rats were surgically monauralized by ablation/obliteration of the cochlea. Auditory brainstem response (ABR) thresholds were determined for ipsilateral and contralateral presentations of 2-kHz, 10-kHz, and 40-kHz toneburst. IAA was calculated by subtracting the frequency-specific ABR threshold obtained from the normal ear from that obtained following tone presentation to the 'dead' ear, and was found to average 65.0 +/- 10.5 dB at 2 kHz, 45.0 +/- 8.4 dB at 10 kHz, and 47 +/- 15.1 dB at 40 kHz (+/- standard deviation). Using data obtained from the animal demonstrating the smallest IAA, masking is not needed until a threshold asymmetry of 50 dB at 2 kHz and 30 dB at 10 and 40 kHz is observed. In order to obtain bilateral auditory threshold information in any animal model of asymmetric hearing loss, data regarding IAA are needed in order to know when to apply contralateral masking and therefore avoid crossover stimulation of the non-test ear. The protocol presented herein provides guidelines for use in any animal model of sensorineural hearing loss which may demonstrate unilateral or asymmetric deficits. PMID- 9390804 TI - Effect of trial-by-trial variation in center frequency on detection of interaural temporal and intensitive differences in bands of noise. AB - Sensitivity to interaural temporal disparities (ITDs) and interaural intensitive disparities (IIDs) was measured as a function of duration in conditions where the center frequency (CF) of the stimuli was constant and, separately, in conditions where the CF of the stimuli was varied on a trial-by-trial basis. Stimuli were bands of noise centered at either 300, 1200, 2400, or 4800 Hz with the bandwidth of the noise being 40% of their CF. The largest effects of shortening duration and varying center CF of the stimuli were degradations of sensitivity to ITDs for stimuli centered at 2400 or 4800 Hz. Overall, the data confirm and extend previous findings which indicate that ITDs and IIDs are processed separately within the central nervous system. PMID- 9390805 TI - A new method of measurement of most comfortable loudness. AB - A new computer-assisted method for the measurement of most comfortable loudness (MCL) is presented, where not only MCL but also its reliability can be graphically displayed and measured. Two unique features concerning the reliability of MCL measurement are obtained in this method: sharpness and stability. Sharpness is defined by the rate of repeat selection of the same level as MCL, and stability is used to express the consistency of a subject's response during the experiment. A method of analyzing sharpness and stability was explained by showing two contrasting cases. Based on the graph configurations, the subjects were divided into two major categories, type A and type B, defined by sharpness of judgment. Type A subjects' judgments are sharper than those of type B. Decreasing the frequency increased the occurrence of type A subjects. Retest results revealed that this method has a high degree of reproducibility of MCL judgment, which supports the reliability of this method. PMID- 9390806 TI - Cerebrospinal fluid loss and threshold changes. 1. Hearing loss in the contralateral ear after operation for acoustic neuroma: an analysis of the incidence, time course, frequency range, size and pathophysiological considerations. AB - In a prospective study, 60 patients who underwent surgery for unilateral acoustic neuromas had the hearing on the contralateral ear tested before and several times after surgery. In 40 patients, a threshold increase was found during the following 9 days. The changes were greatest in the low frequencies immediately after surgery, but after 1 week the treble also became involved. After 3 months, the hearing was normalized. The elder patients more than 50 years of age were more often affected, whereas sex, tumor size, surgical approach or duration of surgery had no influence. The pathophysiological mechanisms are discussed and an intracochlear fluid dysfunction caused by the loss of cerebrospinal fluid is suggested. PMID- 9390807 TI - Cerebrospinal fluid loss and threshold changes. 2. Electrocochleographic changes of the compound action potential after CSF aspiration: an experimental study. AB - Hearing loss has been reported following leakage of cerebrospinal fluid (CSF). The etiology has been attributed to an induced imbalance in the intracochlear hydrodynamics. The present study reports a guinea pig model with surgically induced loss of CSF. In 18 anesthetized animals, CSF was drained by a suboccipital incision in the dura: Eighteen animals were used as controls. The compound action potentials were recorded by an ear canal silver electrode. The animals with CSF loss showed a small increase in threshold and latency, while the control group was unchanged. The concept of an induced inner ear fluid dysfunction after CSF leak is supported by these findings. PMID- 9390808 TI - Cochlear implant place psychophysics 1. Pitch estimation with deeply inserted electrodes. AB - Numerical estimation of pitch was performed by 8 adult subjects implanted with cochlear prostheses manufactured by Cochlear Limited. The electrode arrays had been inserted into the scale tympani to between one and one and a half turns of the cochlea. Using bipolar stimulation, the pitch estimates for each subjects showed an overall reduction with insertion depth of the stimulated electrode. However, for several subjects, after decreasing regularly for the more basal electrodes, pitch estimates showed an abrupt decrease, followed in some cases by a region of low pitch. Two of the subjects, implanted with a modified electrode array, the '20 + 2' which allowed monopolar in addition to bipolar stimulation, exhibited an abrupt decrease in pitch estimate with bipolar but not with monopolar stimulation. In these two subjects, for stimulating electrodes inserted more deeply than about three quarters of a turn, bipolar stimuli produced lower pitch sensations, and presumably more apical neural excitation patterns, than monopolar stimuli. PMID- 9390809 TI - Cochlear implant place psychophysics. 2. Comparison of forward masking and pitch estimation data. AB - Results for forward masking and numerical estimation of pitch were compared in a group of 6 adult subjects implanted with cochlear prostheses manufactured by Cochlear Limited. Data were collected for bipolar + 1 stimulation in all subjects, and for stimulation in one other mode, either common ground or monopolar, for all subjects but one. The pitch data show various irregularities and in each case can be seen to be broadly consistent with the corresponding forward masking data. It is shown that a 'centre of gravity' of the forward masking distribution varies with masker electrode in a manner that is qualitatively very similar to the variation of pitch estimate. It is suggested that, while pitch estimation results are consistent with those from forward masking, the latter contain more detailed information that may be useful in understanding intersubject variations in speech comprehension. PMID- 9390810 TI - Factors affecting auditory performance of postlinguistically deaf adults using cochlear implants. AB - A model of auditory performance and a model of ganglion cell survival in postlinguistically deafened adult cochlear implant users are suggested to describe the effects of aetiology, duration of deafness, age at implantation, age at onset of deafness, and duration of implant use. The models were compared with published data and a composite data set including 808 implant users. Qualitative agreement with the model of auditory performance was found. Duration of deafness had a strong negative effect on performance. Age at implantation had a slight negative effect on performance, increasing after age 60 years. Age at onset of deafness had little effect on performance up to age 60. Duration of implant use had a positive effect on performance. Aetiology had a relatively weak effect on performance. PMID- 9390811 TI - Outer hair cell activity is not required for the generation of the forward masking curve. AB - Forward masking of the auditory brainstem response (ABR) was achieved by increasing the time interval from 0 to 12 ms between the masker offset and the probe onset. The forward masking response demonstrated a near linear function with an approximate 3.0-dB increase in masking threshold for every millisecond interval increase in the control guinea pig. The slope of the masking curve at selected frequencies together with the quantification of hair cell loss through the analysis of cochlear surface morphology was studied before and after chemical insult. The intracochlear infusion of sodium salicylate caused an approximately 45-dB threshold shift of the ABR whereas the slope of the forward masking curve was not significantly different from the control values at the tested frequencies (1, 4, and 8 kHz). Systemic kanamycin administration (400 mg/kg body weight for 9 consecutive days) caused a permanent ABR threshold shift of 43-63 dB at 1, 4, and 8 kHz. The slope of the forward masking curve was not significantly different at 1 kHz despite significant outer hair cell loss. The slope of the forward masking curve at 4 and 8 kHz showed significant reductions at the time intervals between 0 and 4 ms. Analysis of the kanamycin-treated cochleae revealed not only significant outer hair cell loss throughout the cochlea but significant inner hair cell and inner pillar cell loss in the basal end of the cochlea. The results suggest that the outer hair cells are not needed for maintaining a normal forward masking curve, whereas the slope of the forward masking curve is sensitive to alterations induced to either the inner hair cells or the inner pillar cells. PMID- 9390813 TI - Differences in forward masking after a temporary and a permanent noise-induced hearing loss. AB - The forward masking curve of the auditory brainstem response (ABR) at selected frequencies together with the quantification of hair cell loss through the analysis of cochlear surface morphology was studied in guinea pigs before and after acoustic trauma resulting in either a temporary or a permanent threshold shift. In the presence of a noise-induced temporary threshold shift, the slope of the forward masking curve was not significantly different from the pre-exposure curve. In contrast, during the acute phase of the permanent threshold shift, the slope of the forward masking curve was significantly reduced compared to the pre exposure value. After a recovery period of 2 weeks, the slope of the forward masking curve from the permanently damaged group returned to nearly normal values despite a persisting ABR threshold shift and significant loss of outer hair cells. The potential for analyzing the slope of the forward masking curve in order to distinguish between the acute phase of a permanent threshold shift and a temporary threshold shift is discussed. PMID- 9390812 TI - Forward masking is dependent on inner hair cell activity. AB - The goal of this study was to test the hypothesis that the inner hair cell complex (inner hair cell and dendritic contacts) is solely responsible for generating the slope of the forward masking curve. To test this hypothesis two experiments were performed. The first was to measure forward masking from the Bronx waltzing mouse, a mutant possessing an inner hair cell defect. The Bronx waltzing mouse demonstrated an approximately 60-dB auditory brainstem response (ABR) threshold shift compared to CBA/CBA mice at 8 and 12 kHz. The slope of the forward masking curve was significantly reduced compared to the control group, particularly at the early delay times between 0 and 4 ms. The second model employed kainic acid to affect the dendrites beneath the inner hair cell. After the intracochlear infusion of kainic acid, there was an approximately 47-dB ABR threshold shift at 4 and 8 kHz compared to pre-infusion thresholds. The slope of the forward masking curve from the kainic-acid group was significantly reduced compared to the artificial-perilymph group. Primarily the early delay times were affected by kainic acid (0-4 ms). Morphological analysis showed that there was extensive swelling of the afferent nerve radial dendrites under the inner hair cells. The results from the present study, as well as the preceding article, suggest that the analysis of the slope of the forward masking curve may be used for the detection of inner hair cell or radial dendrite damage, independent of outer hair cell damage. The present finding could provide a useful means of employing a clinical test for determining the function of the inner hair cell complex using a non-invasive measure of auditory function. PMID- 9390814 TI - Stability of efferent-mediated protection against acoustic overexposure with long maintenance under barbiturate anaesthesia. AB - When anaesthetized animals are maintained over a long period, crossed-cochlear suppressive and enhancement-in-noise effects mediated by the olivocochlear bundle (OCB), as well as some OCB neuronal responses, show time-dependent variations. The present study determined if there were any such changes in OCB-mediated crossed-cochlear protection against compound action potential (CAP) threshold losses caused by a standard loud sound exposure at 11 kHz, presented under conditions either not evoking OCB-mediated protection (i.e. monaural exposure) or evoking protection (binaural exposure). Maintaining animals for periods up to approximately 30 h from initial anaesthetization resulted in non-significant changes in pre-exposure CAP thresholds. There were also only small changes over select frequency ranges in threshold losses caused by the monaural or binaural loud sound, after a single exposure as well as when the testing of OCB function was extended to examine effects after dual successive exposures, the latter result being determined by application of a previously described additivity model. The features of OCB-mediated protection also showed good stability over the long maintenance. These results are discussed as providing further circumstantial evidence that protection is mediated by a different OCB subcomponent to that/those responsible for other OCB-mediated crossed-cochlear effects. In general, the results show that the barbiturate anaesthetic used here does not significantly modulate the crossed-cochlear OCB effect of protection, even though it has been shown elsewhere to significantly depress other crossed cochlear OCB effects. PMID- 9390816 TI - Research in otology. PMID- 9390815 TI - Detection of the acoustic reflex below 80 dB HL. AB - A new method for detecting the acoustic reflex that utilizes standard otoacoustic emissions recording techniques is introduced and discussed. Two successive identical tone bursts of 100 ms duration and 10 ms interstimulus interval are presented in the occluded ear canal at a repetition rate of one per second. If the acoustic reflex is elicited, the contraction of the stapedius muscle is delayed with respect to the onset of the first stimulus. Hence, the acoustic compliance in the ear canal decreases primarily during the second stimulus. The difference of the microphone signals produced by the two stimuli is computed and averaged across a certain number of repetitions of the sequence. The presentation level is increased until this difference is larger than -40 dB (with respect to the stimulus level) and if its signal-to-noise ratio exceeds 20 dB. For normal hearing subjects, the acoustic reflex threshold measured with this method is on average 8 dB lower than in a standard clinical setup. In 5 out of the 10 tested hearing-impaired subjects, the new method could detect an acoustic reflex at one or more frequencies where no reflex was detected in the clinical setup. PMID- 9390818 TI - In vivo gene transfer into the embryonic inner ear using retroviral vectors. AB - Retrovirus-mediated gene transfer holds great promise for elucidating key genes in the development and function of the inner ear. Retroviral vectors offer a number of advantages over other gene transfer methods including stable and efficient integration into the host genome, high levels of transcription and restriction of expression to a target area. Because of the wide variety of recombinant retroviral vectors currently available, this review outlines which vectors are appropriate for particular applications. Successful strategies for infecting the ear are reviewed and current drawbacks and future directions are discussed. PMID- 9390817 TI - Paradigms and paradoxes: mouse (and human) models of genetic deafness. AB - Mouse models have proved valuable tools in the analysis of human genetic disorders. The identification of the genes mutated in classical mouse mutants and the analysis of the phenotype of mutants following targeted gene disruption have provided some clarification of the development and functioning of the inner ear. A number of these genes also play a role in human deafness. Analysis of mutations in both human and mouse deafness genes has identified a number of distinct phenomena that contribute to the observed phenotype. PMID- 9390819 TI - Production of conditionally immortalised cell lines from a transgenic mouse. AB - This review describes the H2kbtsA58 transgenic mouse (Immortomouse) and its application to the production of conditionally immortalised cell lines from sensory epithelia within the mammalian inner ear. Established cell lines should overcome many of the technical difficulties associated with experimental procedures in auditory and vestibular research. These include the limited amount of tissue available and the relatively complex and laborious dissection. Conditional immortalisation should also allow essential studies on the molecular and cellular mechanisms that govern both the differentiation of sensory cells and the development of sensory epithelia. PMID- 9390820 TI - Transcription factors in the ear: molecular switches for development and differentiation. AB - In order to understand the molecular events underlying differentiation and development in the inner ear, we need to identify and characterize the molecular 'switches' involved in the regulation of gene expression in the system. The most important molecular regulators are represented by a family of proteins generically called transcription factors. This article reviews our current knowledge of the expression of several transcription factors in the ear and their implications for both development and homeostasis in the auditory organs. PMID- 9390821 TI - Cellular commitment and differentiation in the cochlea: potential advances using gene transfer. AB - The development of individual cells as hair cells and supporting cells is a key step during the embryonic formation of the auditory system. However, at present the factors that play a role in the commitment and differentiation of cells as hair cells and supporting cells have not been identified. Recent advances in molecular biological techniques have led to the identification of candidate genes that may be involved in hair cell and supporting cell development, however it has been difficult to determine the specific effects of these genes. The development of new methods for gene transfer into post-mitotic cells should provide powerful new techniques for examining the specific effects of candidate genes. Virally mediated vectors, such as adenovirus and herpes simplex virus, and non-virally mediated vectors, such as lipofectins and biolistics, have been shown to efficiently transfer candidate genes into many different cell types, including hair cells, supporting cells, and spiral ganglion neurons. In addition, studies in other developing systems have demonstrated that these techniques can be used to determine the effects of expression of candidate genes during the specification of individual cell phenotypes. These results suggest that these vectors can be used effectively to study the role of specific genes during the development of the auditory system. PMID- 9390822 TI - The molecular biology of hair cell regeneration in the avian cochlea. AB - The sensory cells of the ear, the hair cells, are damaged by loud noise or certain types of drugs. In the bird cochlea, new hair cells are produced to replace those that are lost. Regeneration also occurs in the vestibular epithelia of birds, fish, and mammals but does not occur in the mammalian cochlea. In order to further our understanding of the regeneration process in the bird cochlea, we have begun to identify the genes that are involved. However, the small size of this organ has made it difficult to use traditional molecular biology methods to address these problems. Recently, many molecular techniques have been adapted for use with small amounts of tissue. Northern blot analysis, the ribonuclease protection assay, semiquantitative PCR and differential display of mRNA are all techniques that are being used to greatly improve our understanding of hair cell regeneration and may eventually provide the information necessary to induce regeneration in hearing-impaired humans. PMID- 9390823 TI - Analysis of gene expression in the organ of Corti revealed by single-cell RT-PCR. AB - Many genes encoding proteins which are expressed in the auditory periphery have been identified in the last years. With single-cell reverse transcription polymerase chain reaction (RT-PCR), the molecular analysis of gene expression can be done on the single-cell level. Furthermore a single-cell RT-PCR experiment can be combined with the electrophysiological characterization of an individual cell. The combination of these two methods will lead to a better understanding of how functional properties of neurons are controlled by the expression of complex proteins. PMID- 9390824 TI - Molecular analysis of excitatory amino acid receptor expression in the cochlea. AB - Our present understanding of excitatory neurotransmission has expanded enormously in the last decade through the use of molecular biology. In the mammalian cochlea, the analysis of excitatory amino acid receptor expression by the reverse transcription-polymease chain reaction (RT-PCR), in situ hybridization and immunochemistry has provided considerable evidence for glutamate as the afferent neurotransmitter. Using these molecular techniques, the ionotropic alpha-amino-3 hydroxy-5-methyl-4-isoxazolepropionate (AMPA), kainate, N-methyl-D-aspartic acid (NMDA) and delta receptor subunits and the metabotropic glutamate receptors have all been detected in the cochlea, in either the spiral ganglion neurons, the hair cells or both. Due to the utility of the techniques and the diversity of expressed neurotransmitter receptors, molecular biology will continue to provide important information for researchers of the auditory periphery. PMID- 9390826 TI - Age-dependent effects of the onset of a conductive hearing loss on the volume of the cochlear nucleus subdivisions and the expression of c-fos in the mongolian gerbil (Meriones unguiculatus). AB - A monaural conductive hearing loss was induced by interrupting the chain of the middle ear ossicles on the right side in gerbils of four different age groups (P12-14, P20-21, P42 and P84). The volumes of the cochlear nucleus subdivisions and the number of cells that expressed immunoreactivity for c-fos after noise stimulation were determined on the left and right side in the deprived animals, and in undeprived control animals when they reached the age of 6 months. The anteroventral cochlear nucleus on the deprived side was reduced in volume when the deprivation started before the age of 3 months. The other cochlear nucleus subdivisions showed no systematic age-dependent reductions. The expression of c fos in the dorsal cochlear nucleus appeared more resistant to a hearing loss, with deprivation being more effective in younger animals. c-fos expression was also dramatically reduced in the ventral cochlear nucleus, regardless of age at the onset of hearing loss. PMID- 9390825 TI - Cholinergic and purinergic neurohumoral signalling in the inner ear: a molecular physiological analysis. AB - The ability to identify the expression of the protein subunits which assemble to form ionotropic receptors for acetylcholine and extracellular adenosine 5' triphosphate (ATP) in individual cells of the inner ear provides examples of the high resolution and exquisite sensitivity which molecular biology brings to the study of hearing and balance. The data from these studies provide both fine detail with respect to the classification of the elements involved and an overview of the sites of potential interaction of both extracellular and intracellular signalling pathways. The high sensitivity necessitates a molecular physiological approach when using these techniques so that these data on the site and extent of expression can be balanced against functional significance. With the demonstration of expression of the alpha 9 subunit of the nicotinic acetylcholine receptor in cochlear outer hair cells, molecular biology has provided an explanation for the unusual cholinergic receptor pharmacology of the olivocochlear efferent innervation which has confounded investigators for decades. In addition, a role for extracellular ATP as a signalling molecule regulating electrochemical gradients and neurotransmission within the inner ear is supported by the extent of P2 receptor expression in this tissue, data which beg for intense functional study. PMID- 9390827 TI - Remote masking in normal-hearing and noise-exposed chinchillas. AB - Remote masking (RM), the phenomenon whereby an intense high-frequency masking noise elevates thresholds for low-frequency signals, has been shown to be sensitive to various types of hearing loss in humans. We performed two experiments to evaluate the chinchilla as a model of RM and to examine changes in RM associated with temporary threshold shifts (TTSs) induced by low-frequency noise exposure. Thresholds for 0.5-, 1- and 2-kHz tones were measured in quiet, then in the presence of a narrow-band (300-Hz-wide) masking noise centered at 3 kHz. In Experiment I, effective masking was measured as a function of masker level, from 48 to 98 dB sound pressure level (SPL; referenced to 20 microPa), to determine threshold and rate of growth of RM in the chinchilla. In Experiment II, RM was measured before, during and after exposure to a low-frequency noise known to produce TTSs in chinchillas (i.e., a 0.5-kHz octave band noise at 90 dB SPL for 6 h/day for 10 days). The results show that normal-hearing chinchillas have the same pattern of RM as humans, and that a noise exposure that produces TTSs also produces rapid and significant changes in RM. PMID- 9390828 TI - A five-generation family with late-onset progressive hereditary hearing impairment due to cochleosaccular degeneration. AB - Cochleosaccular dysplasia or degeneration (Scheibe degeneration) is considered the most common cause of profound congenital hearing impairment, and accounts for approximately 70% of cases 2 with hereditary deafness. A five-generation family with hereditary hearing impairment associated with cochleosaccular degeneration has recently been identified. The diagnosis of classical Scheibe degeneration was based on histopathological findings in the temporal bones of the proband, a 61 year-old profoundly deaf male. Auditory structures in the brainstem of the proband were also studied. Twenty-two members of the family were contacted for surveys and blood samples. Of these, 6 males and 2 females have hearing impairment. Complete audiological evaluation was done on 12 family members, and prior audiologic records of the proband and affected family members were available for study. Affected family members suffer a mild bilateral high frequency hearing loss during childhood and adolescence, and progress to moderate to-profound deafness in the second and third decades of life. The family is suitable for linkage analysis and does not map to previously reported loci harboring autosomal dominant, nonsyndromic hereditary hearing impairment genes. The genetic study of this family will be helpful in identifying the genes which, when mutated, result in Scheibe degeneration. PMID- 9390830 TI - Harold Frederick Schuknecht. 1917-1996. PMID- 9390829 TI - Early damage in the chinchilla vestibular sensory epithelium from carboplatin. AB - Carboplatin, a second-generation platinum drug used in the treatment of cancer, can damage the hair cells in the vestibular system; however, little is known about the time course of its vestibulotoxic effects. The present study examined the acute vestibulotoxic effects of carboplatin (50 mg/kg) in the chinchilla. The duration of the nystagmus response evoked by cold caloric stimulation was significantly reduced 6 h following carboplatin treatment and showed a maximum, permanent reduction of approximately 50% by 24 h after injection. Light microscopic observations at 6 h subsequent to injection revealed swollen afferent dendrites beneath type-I hair cells and the appearance of small vacuoles within the type-I hair cells; these changes were most pronounced in the crista ampullaris of the semicircular canals compared to the maculae of the utricle and saccule. Many mitochondria were swollen and partially depleted of their membranous infoldings. The mitochondrial abnormalities tended to be somewhat more severe in the hair cells than in their afferent terminals. The structural abnormalities in the mitochondria were more severe at 24 h following injection resulting in the appearance of larger and more numerous vacuoles in the hair cells. By 3 days after injection, many type-I hair cells were filled with large vacuoles which often caused severe distortion of the nucleus and disruption of the plasma membrane. Small vacuoles were occasionally observed in type-II hair cells, mainly in the crista ampullaris. These results indicate that the vestibulotoxic effects of carboplatin occur quite rapidly and cause significant disruption of the mitochondria in hair cells and their afferent terminals. PMID- 9390831 TI - Progression of cochlear and retinal degeneration in the tubby (rd5) mouse. AB - Mice homozygous for a defect of the tub (rd5) gene exhibit cochlear and retinal degeneration combined with obesity, and resemble certain human autosomal recessive sensory deficit syndromes. To establish the progressive nature of sensory cell loss associated with the tub gene, and to differentiate tub-related losses from those associated with the C57 background on which tub arose, we evaluated cochleas and retinas from tub/tub, tub/+, and +/+ mice, aged 2 weeks to 1 year by light and electron microscopy. Cochleas from mice of all three genotypes show progressive inner (IHC) and outer hair cell (OHC) loss. Relative to tub/+ and +/+ animals, however, tub homozygotes show accelerated OHC loss, affecting the extreme cochlear base (hook region) by 1 month, and the apex by 6 months. IHC loss in tub/tub animals is accelerated in the basal half of the cochlea, affecting the hook region by 6 months. Spiral ganglion cell losses were observed only in tub/tub mice, and only in the cochlear base. Retinas of tub/tub mice are abnormal at maturity, exhibiting shortened photoreceptor outer segments by 2 weeks, and progressive photoreceptor loss thereafter. Because the tub mutation causes degeneration of sensory cells in the ear and eye but has no other neurological effects, tubby mice hold unique promise for the study of human syndromic sensory loss. PMID- 9390832 TI - Thresholds of intracranially recorded auditory field potentials in the pigeon compared with compound action potential thresholds. AB - The compound action potential (CAP) thresholds provide a reliable indicator for cochlear functional integrity during experimentation in birds as well as in mammals. However, if experimental manipulations are necessary in the middle ear/inner ear spaces, the round window electrodes are often inconvenient. In search for an alternative for CAP recordings, intracranial recordings of acoustically evoked field potentials from the nucleus angularis/magnocellularis were made in pigeons using stereotactically placed electrodes. The responses were compared with those recorded from intracranial surface electrodes placed on the dura mater and compared with CAP responses recorded from the round window. The field potentials recorded from the nucleus angularis/magnocellularis contain a significant contribution from the auditory nerve, as large in amplitude as the CAP recorded at the round window. The recordings from the intracranial surface electrodes were noisier and the contribution from the auditory nerve was too small to be used as a fast monitor of the condition of the inner ear. Threshold curves as a function of frequency could be determined with an automated method from the nucleus angularis/magnocellularis with the same sensitivity and accuracy as from the round window CAP within a few minutes. These results demonstrate that stereotactic recordings of field potentials from the nucleus magnocellularis/angularis region are a suitable alternative to reliably monitor the condition of the inner ear when round window electrodes cannot be used. PMID- 9390833 TI - Attenuation of salicylate-induced tinnitus by Ginkgo biloba extract in rats. AB - The effects of an extract from Ginkgo biloba, EGb 761, on tinnitus were tested using an animal model of tinnitus. Daily oral administration of EGb 761 in doses from 10 to 100 mg/ kg/day began 2 weeks before behavioral procedures and continued until the end of the experiment. Tinnitus was induced by daily administration of 321 mg/kg sodium salicylate s.c. (corresponding to 275 mg/kg/day of salicylate acid) in fourteen groups of pigmented rats, 6 animals/group. The results from salicylate- and EGb-761-treated animals were compared to control groups receiving either salicylate, saline, or EGb 761 only in doses of 100 mg/kg. Administration of EGb 761 resulted in a statistically significant decrease of the behavioral manifestation of tinnitus for doses of 25, 50 and 100 mg/kg/ day. PMID- 9390834 TI - Arrangement of vestibular nerve fibers in the semicircular canal crista of the chinchilla. AB - The topographic arrangement of vestibular nerve fibers innervating semicircular canal cristae of the chinchilla was studied using computer-aided video-microscopy and three-dimensional reconstruction. At the level 20 microns proximal to the base of the crista, bundles consisting of 30-50 nerve fibers each were identified. Nerve fibers in bundles were classified into seven categories depending on the diameter. We confirmed that large nerve fibers were more frequently found in the central bundles and small nerve fibers were more frequently found in the peripheral bundles. The central bundle might function as a physiological unit coding various types of head movements, whereas the peripheral bundle might contribute more to the detection of slow and long-lasting movements giving rise to tonus and posture changes. The canalicular nerve may code rotational acceleration of the head via function- and locus-specific nerve fiber bundles. PMID- 9390835 TI - Cochlear potentials in clinical audiology. AB - The recording of cochlear and auditory nerve potentials in humans via Electrocochleography (ECochG) has emerged as a valuable tool for a variety of clinical applications. This review consolidates current research on the use of cochlear potentials and ECochG in the clinical setting and identifies several areas in need of additional study. Methodological topics discussed include a review of ECochG recording approaches (i.e. transtympanic versus extratympanic) and issues related to choice of stimuli (clicks versus tonebursts). The review of current applications for cochlear potentials focuses primarily on the use of ECochG in the identification and treatment of Meniere's disease/endolymphatic hydrops (MD/ELH). Other uses for ECochG also are presented (e.g. intraoperative monitoring, enhancement of ABR wave I, estimation of hearing sensitivity). Several suggestions are offered to maximize the predictive value of ECochG in the diagnosis of MD/ELH. PMID- 9390836 TI - Estimation of the pure-tone audiogram by the auditory brainstem response: a review. AB - This review paper briefly considers how stimulus, noise masking and recording parameters affect the frequency and place specificity of auditory brainstem responses (ABRs) to air- and bone-conducted stimuli. Issues concerning the use of clicks for ABR threshold estimates will first be presented, followed by results for tone-evoked ABR thresholds and how well they predict the pure-tone behavioral audiogram. Noise-masking options (e.g. high-pass noise, notched noise and white noise) to improve the frequency specificity of tone-evoked ABRs, which are now available on clinical ABR units, will also be discussed. The goal of this article is to demonstrate that ABRs to tonal stimuli can be successfully recorded in most clinical environments and can provide reasonably accurate estimates of 500- to 4000-Hz pure-tone behavioral thresholds in infants, children and adults. Specific parameters and protocols for obtaining frequency-specific ABR threshold responses are provided. PMID- 9390837 TI - The N1 response and its applications. AB - Some properties and applications of the N1-P2 complex (100-200 ms latency) are reviewed. N1-P2 is currently the auditory-evoked potential (AEP) of choice for estimating the pure-tone audiogram in certain subjects for whom a frequency specific, non-behavioural measure is required. It is accurate in passively cooperative and alert older children and adults. Although generally underutilized, it is an excellent tool for assessment of functional hearing loss, and in medicolegal and industrial injury compensation claimants. Successful use of N1-P2 requires substantial tester training and skill, as well as carefully designed and efficient measurement protocols. N1-P2 reflects conscious detection of any discrete change in any subjective dimension of the auditory environment. In principle, it could be used to measure almost any threshold of discriminable change, such as in pitch, loudness, quality and source location. It is established as a physiologic correlate of phenomena such as the masking level difference. Thus, N1-P2 may have many applications as an 'objective' proxy for psychoacoustic measures that may be impractical in clinical subjects. Advances in dipole source localization and in auditory-evoked magnetic fields (AEMFs) have clarified the multiple, cortical origins of N1 and P2. These potentials are promising tools for the neurophysiologic characterization of many disorders of central auditory processing and of speech and language development. They also may be useful in direct 'functional imaging' of specific brain regions. A wide variety of potential research and clinical applications of N1 and P2, and considerable value as part of an integrated, goal-directed AEP/AEMF measurement scheme, have yet to be fully realized. PMID- 9390838 TI - Auditory event-related potentials in the study of developmental language-related disorders. AB - This article reviews recent auditory event-related potential (ERP) studies of developmental language disorder (DLD) and dyslexia/reading disorder (RD). The possibility of using ERPs in searching for precursors of these disorders in the early development of infants at risk is also discussed. Differences in exogenous/sensory ERPs at the latency range of P1 and N1-P2 components have been reported between groups with DLD and RD and control groups. Latency differences between the groups may be related to a common timing deficit suggested by some researchers to be one of the possible underlying factors both in DLD and dyslexia. N1 amplitude group differences may be partly related to arousal/attentional factors and partly to the 'tuning' of the auditory sensory system. Mismatch negativity deviations in DLD children seem to indicate differences in sensory memory functions. Differences between the reviewed clinical groups and controls exist also in the endogenous P3 component, though less consistently in DLD children. In both clinical groups the P3 amplitudes are, in general, lower and the latencies longer compared to those in controls. These findings are discussed in terms of possible differences in higher cognitive functions that are not specific to modality. Altered hemispheric asymmetries in DLD and RD children, as compared to controls, are commonly found in many of the reviewed ERP components. Differences in ERPs of DLD and dyslexic children in comparison to controls may not reflect only maturational lag but also more fundamental processing deficiencies. PMID- 9390839 TI - Mismatch negativity--the measure for central sound representation accuracy. AB - The mismatch negativity (MMN), elicited by any discriminable change in a repetitive sound even when this sound is not attended to, provides a pre perceptual physiological measure of the accuracy of the central sound representation in the human brain. This accuracy, which can be measured separately for the different features of the sound, determines the individual's sound discrimination accuracy in normal and various pathological conditions. PMID- 9390840 TI - Towards the possible clinical application of the mismatch negativity component of event-related potentials. AB - The mismatch negativity (MMN) is an event-related potential component that signifies neurophysiological processing of fine acoustic differences. The MMN indicates attention-independent change detection, reflects auditory sensory memory and provides a physiological measure of difference sensitivity. This paper will provide an overview of current research where results gained by MMN testing in different patient groups were central to the interpretation of an assumed abnormality of processing or storing acoustic features. PMID- 9390841 TI - Episodic ataxia type 1 and 2 (familial periodic ataxia/vertigo). AB - Episodic ataxia (EA) is a rare, disabling condition of autosomal dominant inheritance, but it is not a distinct clinical entity. Synonyms are familial periodic ataxia or hereditary paroxysmal cerebellar ataxia. Family members have a similar clinical syndrome; however, the syndrome varies considerably from family to family. At least two groups of disorders have been separated clinically: (1) episodic ataxia type 1 (EA-1), which manifests without vertigo and is associated with 'interictal' myokymia, and (2) episodic ataxia type 2 (EA-2), which often manifests with vertigo and is associated with 'interictal' nystagmus. EA-1 and EA 2 have been identified as channelopathies. EA-1 is due to different heterozygous missense point mutations in a voltage-gated (delayed rectifier) potassium channel gene (KCNA1/Kv1.1) on chromosome 12p13, whereas EA-2 is caused by mutations of the cerebral P/Q-type calcium channel alpha 1 subunit gene CACNL1A4 localized on chromosome 19p, which is highly expressed in the cerebellum. The diagnosis of EA 1 and EA-2 is important, since they can be easily treated and are often mislabeled. As effective as acetazolamide is in preventing attacks, prospective studies still have to prove whether it can prevent progressive ataxia in EA-2 or even improve chronic cerebellar deficits. PMID- 9390842 TI - Limits of normal for pressure sensitivity in the fistula test. AB - In patients with perilymphatic fistula (PLF), nystagmus may sometimes be elicited by application of pressure to the external ear canal. The extent to which the normal population also exhibits such 'pressure sensitivity' is presently unknown. Our goal was to determine the limits of normal pressure sensitivity and to quantify the performance of the fistula test. Our subjects consisted of 13 normal controls and 7 patients with a history of pressure sensitivity who later underwent exploratory tympanotomy. We measured nystagmus prior to and following pressurization of the external ear canal. Pressure was applied manually over 60 s with a pneumatic otoscope bulb. In normal subjects, change in nystagmus between prepressure and postpressure tests ranged from -1.3 to 0.9%s. In patients, change in nystagmus greater than the 95th percentile limits of normal was not a reliable indication of PLF. PMID- 9390843 TI - Transtympanic electrocochleography in the assessment of perilymphatic fistulas. AB - An objective method for the pre-operative diagnosis and the post-operative assessment of a presumed perilymphatic fistula (PLF) using transtympanic electrocochleography is presented. Three cases are reported in which the history of the disease and the symptoms strongly suggested the presence of a PLF. Pre operative transtympanic electrocochleography (TT ECoG) recordings at rest showed changes similar to those of endolymphatic hydrops and signs of instability of the inner ear hydrodynamic system during raised intrathoracic pressure. Surgery revealed a visible leak in two of the three cases. Both windows were repaired in all the patients. All patients were relieved from their vestibular symptoms at the time when the post-operative TT ECoG was conducted 3-6 months later. The post operative recordings were stable during raised intrathoracic pressure and the previously elevated summating potentials decreased which was interpreted as an objective indication of the recovery of the hydrodynamic system. However, later one of the patients again developed recurrent vertigo. Twenty patients with well documented Meniere's disease were used as a control group. TT ECoG was conducted during raised intrathoracic pressure. The Meniere patients showed stable recordings. It is suggested that among patients with suspected PLF and signs of hydrops in TT ECoG, a dependence on the intrathoracic pressure reflected in the recordings may indicate a possible fistula. PMID- 9390844 TI - Tinnitus and translabyrinthine acoustic neuroma surgery. AB - The purpose of this investigation was to study the effects of translabyrinthine acoustic neuroma surgery on tinnitus in a consecutive sample of patients operated on between 1988 and 1994 in Uppsala (Sweden). A postal questionnaire was returned by 141 patients, yielding a 90% response rate without reminder. The results showed that tinnitus was experienced by 70% of the patients before surgery and 60% after surgery. In general, low degrees of tinnitus distress were found, which was confirmed by the questionnaire results. Ratings of tinnitus distress after surgery, using the Klockhoff and Lindblom grading system, showed that 48% had tinnitus of grade I, 46% of grade II, and 6% of grade III. Pre- and postsurgery grading of distress did not change significantly. There was a 35% risk for developing tinnitus when no preoperative tinnitus was present and a 15% chance that tinnitus disappears when present preoperatively. PMID- 9390845 TI - Surgical strategy of cochlear implantation in patients with chronic middle ear disease. AB - We report 10 postlingually deafened adults in whom the electrophysical criteria for cochlear implant were fulfilled, except that they showed the following unfavorable middle ear lesions: otitis media with effusion, chronic perforative otitis media, cholesteatoma and previous radical ear operation. Staged operations for cochlear implant were performed in 8 cases, and 2 patients who had undergone radical ear operation had a single-stage operation. As a first step, one of the following was performed in each patient as surgically indicated: myringoplasty with or without mastoidectomy, mastoidectomy with reconstruction of the posterior wall of the external canal, mastoidectomy with the insertion of a ventilation tube, radical mastoidectomy or surgical cleansing of the radical cavity. From 6 months to 2.5 years after the first operation, the actual cochlear implant was performed in the second or third stage. There was no major complication as a result of electrode insertion into the cochlea and the results of speech perception in these cases were not different from those in patients with normal middle ears. In our experience, it was considered that the staged operations would enable successful cochlear implants in selected patients with pathological middle ear lesions even if they had previously been diagnosed as contraindicated for this procedure. In a case with radical ear cavity a single-stage operation could be performed when there was no cavity problem. PMID- 9390846 TI - The history of pH and blood gas analysis. AB - It is difficult for today's clinician to appreciate the rapid evolution of technology relevant to pH and blood gas measurements. Discovery of the scientific foundations took three centuries, whereas development of practical methods for clinical application took only three decades from the realization of their potential clinical importance. All indications are that advancement will continue at the same pace for the next several decades. PMID- 9390847 TI - Acid-base balance. AB - This article emphasizes the interactions between the respiratory and metabolic sources of acid and their clearance. It includes the chemistry of acid-based balance as expressed by the Henderson-Hasselbach equation, the principles and importance of biologic buffering systems, and a brief review of renal regulation of acid-based balance. Finally, the reader is given a set of guidelines for a systematic approach to the analysis of acid-base data. PMID- 9390848 TI - Oxygen transport. AB - The most important factor in the development of complex life forms is the ability to deliver oxygen to cells within complex tissues and organs. Hemoglobin plays a significant role in this scheme. Oxygen transport is a complex phenomenon that encompasses the oxygen content, oxygen delivery mechanisms, and oxygen use. All three components must operate appropriately if the organism is to function and survive. PMID- 9390849 TI - Blood gas analyzers. AB - Laboratory-based blood gas analyzers have reliably produced pH, PCO2, and PO2 analyses for several decades. Over this time, their design changes reduced maintenance procedures, microprocessor-controlled functions simplified operation, and regulations governing their use have expanded. This article presents the electrochemical principles of the pH and blood gas electrodes and the operational procedures necessary to satisfy regulatory standards. PMID- 9390850 TI - Co-oximetry. AB - The adequacy of tissue oxygenation depends on the interaction of many factors. Assuming the presence of sufficient tissue perfusion, oxygenation failure must be caused by depressed respiratory efficiency (shunt or other ventilation/perfusion mismatch), inadequate FIO2/PaO2 relationships (alveolar-capillary diffusion deficits, for example), or oxygen transport difficulties (hemoglobin loading/unloading dysfunction). When presented with patients whose respiratory distress is not alleviated by application of increasing levels of FIO2 and seemingly adequate SaO2 values, one must look toward less obvious reasons for the disparity between subjective and objective findings. Early response by clinicians to situations such as those just mentioned should include a survey and analysis of hemoglobin status. It is important to note that meaningful co-oximetry results depend on the quality of the patient history and other laboratory tests to rule out factors that might affect the co-oximetry results. Good preparation of the sample is essential to ensure that adequate hemolysis has occurred and that the sample was not contaminated prior to analysis. A well designed and executed program of preventive maintenance and QA is important. It should include preparation and sampling as well as technique and instrument integrity. All of these are essential for safe, effective, and accurate determination and dissemination of this important clinical information. PMID- 9390851 TI - Temperature correction of blood gas values. AB - The popularity of routine temperature correcting of pH, PCO2 and PO2 values is based on the observation that large differences in the blood gas values are present when the patient's temperature is profoundly hypo- or hyperthermic. This observation leads some clinicians to the unsubstantiated conclusion that uncorrected 37 degrees C values are "wrong." The danger in this superficial thought process is that one might reach the unfounded conclusion that temperature corrected values are "right." The simple truth is: With significant changes in patient temperature, we do not fully understand the complexity of the effects on metabolism, vascular function, and respiration. Both corrected and uncorrected blood gas values, therefore, are of uncertain usefulness in patients with significant deviations in body temperature. There is no logical or scientific basis for the assumption that temperature-corrected values are better than the values obtained at 37 degrees C. In fact, the available technical and biological data lead to the conclusion that, in almost all circumstances, there is no clinical advantage to using values other than those at 37 degrees C. In addition, the routine process of temperature correction of blood gases involves several practical disadvantages. First, interpretation of the corrected values demands deviation from the familiar and well-documented guidelines for interpreting 37 degrees C values. Second, temperature correction assumes the laboratory has received the patient's true temperature at the time of sampling. My experience is that the patient's true temperature often is not reported or is reported erroneously. Third, temperature-corrected values can be confused with uncorrected values and vice versa. Available data support the practice that only uncorrected (37 degrees C) blood gas values should be used and reported routinely. Temperature-corrected values should be calculated only when specifically requested and the onus for clinical use of temperature-corrected values lies with the clinician who requests them. PMID- 9390852 TI - Pulse oximetry. AB - Pulse oximetry is a reliable, noninvasive, easy to use means of continuously determining arterial oxygen saturation in virtually any setting. This article details the historical and technical development of this monitor; reviews the literature on application, accuracy, and response; and presents an overview of future advances. PMID- 9390853 TI - Capnography. AB - Capnography measures exhaled carbon dioxide and is most useful when applied directly to patient care. This is in circumstances of detecting misplacement of the tracheal tube, dysfunction of respiratory apparatuses, detection of abnormal lung function, successful cardiopulmonary resuscitation, and trending of deadspace changes. The least reliable application is to reflect alveolar ventilation (PaCO2). This application is most common during general anesthesia and weaning from mechanical ventilation. Provided the patient has a stable cardiac status, stable body temperature, absence of lung disease, and normal capnogram, PETCO2 monitoring may assist in estimating PaCO2. The use of capnography in patients with severe respiratory failure should be applied with careful reflection. The increased V/Q mismatch that is consistent with a widened P(a-ET) gradient, as well as worsening hypercapnea with increased peripheral carbon dioxide production, can lead to erroneous PETCO2 values. Capnography may be least useful in the sickest patients. PMID- 9390854 TI - Transcutaneous measurement of partial pressure of oxygen and carbon dioxide. AB - Transcutaneous monitoring is noninvasive and relatively simple to use. In neonates and small infants, this monitoring technique may provide very useful clinical information. Transcutaneous gas monitoring, using conventional electrochemical techniques, provides a means of trending the values of PaO2 and PaCO2 in most patients with relatively normal cardiovascular function. In patients with compromised cardiopulmonary function and in many adults, because of different skin structure, transcutaneous gas monitoring will not accurately reflect arterial blood gas tensions. Because transcutaneous gases depend on skin perfusion, however, it may be useful in monitoring tissue perfusion, especially in patients with peripheral vascular disease and tissue flaps. The heating of the monitoring probe necessitates frequent site changes to avoid thermal injury, which make it more labor intensive than other noninvasive monitoring methods. PMID- 9390855 TI - Point-of-care testing. AB - Point-of-care testing refers to testing outside of the central laboratory at or near the patient's bedside. The practice greatly decreases turnaround time for testing and has improved outcome and decreased length of stay in some patient groups. Advances in technology have made analyzers increasingly portable with expanded testing capacities while maintaining standards for accuracy required by regulatory agencies. It is possible for clinicians to perform testing that historically was performed only in the central laboratory by trained laboratory technicians. Determination of all appropriate bedside testing for different clinical areas and patient groups will require further investigation and debate. PMID- 9390856 TI - Blood gas monitors. AB - In vitro blood gas analysis requires limitation of the frequency of serial blood gas measurements for two major reasons--blood loss and cost. In vivo or ex vivo blood gas monitors eliminate these factors because the measurements are available continuously, or as frequently as deemed desirable, without permanently removing blood or imparting additional cost. For patients with arterial catheters in place, the propriety of blood gas monitors is obvious as long as there is no requirement to alter the size, location, or placement of the arterial catheter and the routine use of the arterial catheter system is unaffected. Further, personnel exposure to the patient's blood, and the risk of nosocomial infection from contaminated arterial catheters, should be reduced because the integrity of the arterial catheter and tubing system is not interrupted to obtain blood gas values. Blood gas monitors and point-of-care analyzers should significantly reduce therapeutic decision time (the interval from ordering the test to initiating a therapeutic action based on the test results), thereby enabling rapid titration of common therapeutic modalities such as oxygen administration, positive pressure ventilation, positive end-expiratory pressure, and manipulation of acid-base balance. The transfer of blood gas measurements from laboratory analyzers to the combination of blood gas monitors and point-of-care analyzers should have as profound an impact on acute care medicine as did the introduction of laboratory-based blood gas analysis over 30 years ago. In the current medico economic environment, however, we must be certain that these devices are reliable, consistent, and cost beneficial in order to avoid widespread application of yet another technology that provides more data, greater costs, and only questionable patient benefits. PMID- 9390857 TI - The rising tide of asthma. Trends in the epidemiology of morbidity and mortality from asthma. AB - The increase in the asthma mortality rate seen in the past decade has stimulated much discussion, research, and controversy. The epidemiology of asthma morbidity is reviewed, with special attention to the roles of demographics, socioeconomic status, and iatrogenesis in the rising tide of asthma mortality. PMID- 9390858 TI - Pathophysiology and management of life-threatening asthma. AB - With sound medical management and good patient education, only a small minority of patients with asthma ever experience a life-threatening episode. The pathophysiology, clinical presentation, and management of life-threatening asthma are reviewed. Special emphasis is placed on identification of the fatality-prone asthmatic patient and on avoidance of complications of treatment that significantly add to morbidity and mortality rates. PMID- 9390859 TI - Ambulatory care of the adult asthma patient. AB - Asthma is a chronic airway disorder that is a serious public health problem in many countries. Asthma affects people of all ages. It ranges from mild to severe and sometimes is fatal. Over 100 million people worldwide have asthma and the prevalence is increasing, especially in the cohort aged 1 to 14 years. Risk factors that lead to the development of asthma appear to be largely environmental and therefore potentially preventable. Lifestyle changes and exposure to allergens may be responsible for the increase in prevalence. PMID- 9390860 TI - The role of allergy in asthma. AB - Allergic triggers remain vitally important for a large proportion of asthmatics. Increasing evidence indicates that allergic sensitivity may not only cause ongoing asthmatic symptoms with bronchial hyperreactivity and airway inflammation but may also serve as an inducer of the asthmatic condition. Patients with a history and clinical setting suggestive of allergic sensitivity should be evaluated carefully. Whenever possible, allergen avoidance should be encouraged strongly in allergen-sensitive patients with a strong emphasis on patient education. In those patients who continue to be symptomatic with good avoidance measures and appropriate pharmacotherapy, allergy immunotherapy should be considered as an additional treatment measure. PMID- 9390861 TI - Aerosol therapy. AB - Aerosols are commonly used in the treatment of patients with pulmonary disease. The clinician must choose the appropriate aerosol delivery device. For both spontaneously breathing and mechanically ventilated patients, the first choice of device is usually the metered-dose inhaler. There are important differences among devices in the dosage delivered to the lungs. For the typical prescription, nebulizers deliver more drug to the lungs than metered-dose inhalers, which may be particularly important for acutely ill patients. PMID- 9390862 TI - Bronchial challenge testing. AB - Airways hyperresponsiveness can occur as solitary evidence of airways dysfunction. Because hyperresponsiveness is associated with the presence and severity of disease, it is important to measure and quantitate airways responsiveness. The most useful challenge procedure appears to be the methacholine challenge, not so much because it offers the best specificity of the challenge procedures but because it has been characterized well. Although the procedures are simple, interpretation of the results is problematic. It is important to remember that the key features of asthma are the periodicity and lability of signs and symptoms. Assessment of airways responsiveness is a significant addition to the armamentarium of the modern diagnosis and treatment of asthma. PMID- 9390863 TI - Management of the difficult asthmatic. Gastroesophageal reflux, sinusitis, and pregnancy. AB - The frustration of treating the difficult asthmatic can be minimized by recognizing that interactions of disease processes can complicate therapy. At the same time, therapy of asthma can be simplified by the awareness that control of allied conditions can improve the response to treatment directed at the airways. The future likely will bring more challenges as our ability to cope with complicated asthma patients improves. PMID- 9390864 TI - Diagnosis and management of acute asthma. AB - Deaths from asthma are relatively uncommon but have continued to rise worldwide in the past few decades, despite a better understanding of the disease and an increased number of patient medications. This often is attributed to inadequate assessment and treatment of the disease. This article focuses on defining several items that pertain to acute severe asthma. In addition, the pathophysiology and clinical manifestation of acute asthma, as well as current investigational tools, are reviewed. Traditional and nontraditional therapies of acute asthma are discussed. Finally, complications of severe asthma and the long-term outcome are discussed, with appropriate preventive measures stressed strongly. PMID- 9390865 TI - Mechanical ventilation in asthma. AB - The goal of respiratory support in asthma is similar to the goals in other forms of respiratory failure: to support gas exchange while avoiding complications. The specific respiratory support goals for asthmatics are discussed in this article, as well as recommended ventilator adjustments to meet these goals. PMID- 9390866 TI - Asthma rehabilitation program. AB - Comprehensive management of asthma includes proper use of medication adjustments in patient lifestyle, exercise conditioning, and patient education to maximize self-management capabilities. Pulmonary rehabilitation programs have used these strategies successfully to improve the functional status and reduce the health care costs of patients with chronic obstructive pulmonary disease. Adapting these programs to the asthmatic population is an important and cost-effective goal. PMID- 9390867 TI - The rationale for therapist-driven protocols. AB - Misallocation of respiratory care, defined as ordering therapy unlikely to provide benefit (overordering) and failing to provide beneficial therapy (underordering), is common and provides the rationale for implementing therapist driven protocols. This article reviews the frequency of misallocation of respiratory care and explores possible reasons for misallocation. Therapist driven protocols represent a new model for delivering respiratory care that is designed to minimize misallocation. PMID- 9390868 TI - A short history of therapist-driven respiratory care protocols. AB - The literature of the TDP movement continues to grow, but a legitimate concern regarding mindless acceptance of the new paradigm is now being voiced. As this issue goes to press, it appears clear to this writer that the orderly growth of the TDP paradigm has led to its widespread acceptance. The future will determine whether our early predictions of its success are well founded. PMID- 9390869 TI - Constructing a therapist-driven protocol. Spectrum of types and quality control. AB - Therapist-driven protocols for the implementation and delivery of respiratory care must be tailored to fit the needs of the individual institution. The method that is chosen for constructing a therapist-driven protocol depends on the type of protocol desired (disease-, symptom-, or treatment-based) and on whether a narrative or flow-diagram format is preferred. Because the goal of therapist driven protocols is to provide a systematic method for determining appropriate respiratory care, quality control measures are necessary to ensure that desired outcomes are achieved. Quality control monitoring can be performed through the use of case-study exercises, verbal shift reports, and care-plan audits. Results of quality control monitoring techniques can be used to guide modification of protocols. PMID- 9390870 TI - Implementing therapist-driven protocols. AB - In January of 1993, as part of a hospital-wide cost-reduction strategy, the University of California San Diego (UCSD) Medical Center Respiratory Care Department implemented a patient-driven protocol program designed to utilize the assessment skills and judgments of respiratory care staff, within physician approved guidelines. This program produced a 60% reduction in the use of hand held nebulizer therapy and chest physical therapy in the institution, with a substantial decrease in operational expenses. This article describes key elements of the implementation of protocol-driven programs, provides examples from the UCSD experience, and offers insights gained from others who have been successful agents of change. It describes patient-driven protocols, how they can be implemented, the barriers to and promoters of such protocols, and what the results can be for a respiratory care department. PMID- 9390871 TI - Assessment instruments in respiratory care. AB - The basis for using therapist-driven protocols effectively is an accurate assessment of the patient's respiratory status. Patients must be assessed to initiate indicated therapy and reassessed so that therapy can be modified or discontinued if no longer needed. This article addresses the role of respiratory therapists in patient assessment for the selection of appropriate and effective protocols. It also describes the use of a systematic and consistent process for patient assessment. PMID- 9390872 TI - Therapist-driven protocols for adult non-intensive care unit patients. Availability and efficacy. AB - As the growth of TDPs increases throughout the country, the need to evaluate the efficacy of TDPs becomes paramount. For TDPs to become a standard of practice they must respond adequately to their intended purpose. TDPs must reduce the amount of misallocation of respiratory services without compromising quality care. As the reduction of misallocation occurs, the results should show a reduction in department and patient costs. Protocols are not new to the medical community, but their need and use for respiratory care has increased. Preliminary studies thus far suggest that the use of TDPs can lessen misallocation of care without adverse events, and with the use of TDPs the number of respiratory modalities and the cost associated with these therapies have decreased. Further examinations of the efficacy of TDPs must be ongoing, however, in order to ensure that these preliminary studies truly reflect the outcomes of the use of TDPs. PMID- 9390873 TI - Therapist-driven protocols in adult intensive care unit patients. AB - TDPs occasionally are used to standardize or control respiratory management of the critically ill. Weaning protocols are most common. Little objective evaluation of the effects of TDPs in the critically ill has been published. Most protocols have been developed to improve efficiency of respiratory care staff and reduce unnecessary treatments in non-ICU patients. The most important reason for using TDPs in the ICU is to improve consistency of care. Reduction of variation between individual therapist style improves physicians' trust in the respiratory care department. Improved consistency may allow novel ICU therapies to be evaluated objectively. In general, TDPs are not directly transportable from one area or institution to another. TDPs require local development, and all interested parties must be part of the development process for success. The process of creating TDPs provides a forum for physicians, nurses, and therapists to establish mutual respect and understanding. The analytic approach needed to create useful TDPs provides a critical evaluation of unit procedures and promotes changes in care delivery extending outside the TDP. The complexity of disease process and patient care in the ICU makes comprehensive TDPs difficult to establish; however, use of computers for decision support can overcome the limitations of paper flow charts. Even without comprehensive TDPs, the development process is important to improving and understanding care of the critically ill. The effects of TDPs on ICU patient outcome are unknown currently. Benefits are possible and improved collaboration, better respiratory care staff morale, consistency of approach to care, and critical approach to clinical decision making can be gained by attempting to develop TDPs for respiratory care delivery in the ICU. PMID- 9390874 TI - Therapist-driven protocols for pediatric patients. AB - Most therapist-driven respiratory care protocols deal with adult care. The greatest difficulty we have encountered when implementing pediatric protocols involves patient assessment. We have found that with any protocol the key factor is to monitor closely the result of the treatment and to analyze that outcome and compare it with the purpose of the therapy. When careful clinical assessments are accomplished, pediatric protocols can be established. PMID- 9390875 TI - Legal aspects of therapist-driven protocols. Do therapist-driven protocols place therapists in a legally compromising position? AB - The recent introduction of therapist-driven protocols has given the appearance of restricting the professional judgment of respiratory therapists with decision tree robotics while contemporaneously catapulting them into the practice of medicine. This is happening in the midst of a spiraling litigation climate. This article examines the legal aspects--from malpractice to licensure--of this exciting new practice known as therapist-driven respiratory protocols. PMID- 9390876 TI - Therapist-driven protocols and newer models for patient care delivery. AB - There continues to be increased scrutiny regarding the quality and cost effectiveness of health care delivery in this country. As stated previously, new models of patient care delivery are designed to streamline and simplify hospital procedures in order to enhance this quality and cost efficiency. There is evidence that many of the elements of patient-focused care are adding value to the quality and efficiency of care. Evidence also indicates that therapist-driven protocols offer a promising solution to the misallocation of respiratory care, which is now widespread. The greatest cost savings to hospitals, it has been shown, comes from the elimination of unnecessary care, which therapist-driven protocols are designed to reduce. In the bigger picture of patient care delivery, however, more extensive and critical evaluation is needed before hospitals commit large sums of money to redesign their infrastructures. In other words, it is not yet proved conclusively that new models of patient care delivery are enhancing quality while cutting costs. To date, there is little documentation by hospitals of improved service or financial outcomes associated with new models of patient care delivery. It is likely that decentralizing ancillary services, streamlining procedures, and cross-training employees can help hospitals improve patient satisfaction and staff efficiency. In fact, there is evidence that the cost savings potential for hospitals that implement employee redeployment and cross training can be significant. Although patient-focused care is worthwhile primarily as an effort to simplify care delivery, the biggest cost savings comes from eliminating unnecessary care entirely. Although the evidence does not yet support hospitals' taking a major plunge into patient-focused care, it has been shown that many elements of patient-focused care are worth implementing and can be done inexpensively. Inexpensive activities that are likely to produce meaningful cost and quality gains include cross-training employees in bedside ancillaries, redeploying medical records, redeploying admitting, redeploying support services, and adding automated drug and supply dispensing to nursing units. Hospitals can minimize their financial risks and still achieve many of the benefits of patient-focused care by taking a middle-of-the-road approach to restructuring and redesign. PMID- 9390877 TI - A historical perspective on the use of noninvasive ventilatory support alternatives. AB - This article traces the development of mechanical ventilatory support methods from the use of body ventilators to tracheal cannulation to the use of noninvasive ventilatory support and airway secretion management alternatives. Although it has been known that tracheostomy tubes could be used for ventilatory support and airway secretion management since 1869, body ventilators continued to be the main methods of long-term ventilatory support in the United States, with tracheostomy performed only for patients with severe bulbar muscle dysfunction, until the late 1950s. Recent technological developments, however, have created renewed interest in noninvasive alternatives. PMID- 9390878 TI - Noninvasive positive pressure ventilation. Equipment and techniques. AB - Successful application of noninvasive positive pressure ventilation is largely dependent on available equipment and the approaches used to apply it. Third generation intensive care unit ventilators and portable volume and pressure ventilators may be used for noninvasive positive pressure ventilation. A variety of facial interfaces currently are manufactured, and all should be available. A well-trained therapist with available time is the final ingredient for successful use of noninvasive positive pressure ventilation. PMID- 9390879 TI - Body ventilators. Equipment and techniques. AB - Body ventilators have been used since the late 1800s and are still used today. This article reviews all of the body ventilators available today including tanks, cuirasses, wraps, rocking beds, and intermittent abdominal pressure ventilators. Diaphragm pacers and glossopharyngeal breathing also are reviewed. Clinical application of the ventilators, initiation, patient monitoring, and follow-up are reviewed. PMID- 9390880 TI - The use of noninvasive respiratory muscle aids in the management of patients with progressive neuromuscular diseases. AB - Most patients with primarily ventilation impairment can use noninvasive alternatives to tracheostomy for long-term ventilatory support and airway secretion management. The most important and versatile method of noninvasive ventilatory support is mouthpiece intermittent positive pressure ventilation. Mouthpiece intermittent positive pressure ventilation also can be used via a Lipseal for nocturnal support. The intermittent abdominal pressure ventilator is an option for daytime aid, and nasal intermittent positive pressure ventilation is usually preferred for nocturnal support. Manually assisted coughing and mechanical insufflation-exsufflation can be critical for airway secretion elimination. PMID- 9390881 TI - Long-term noninvasive ventilation for patients with thoracic cage abnormalities. AB - Long-term noninvasive ventilation offers the patient with thoracovertebral deformities, including deformities that result from the severe skeletal and chest wall sequelae of tuberculosis, what long-term oxygen therapy has offered patients with chronic obstructive pulmonary disease: improved survival and prevention or alleviation of cor pulmonale. Long-term noninvasive intermittent positive pressure ventilation, particularly nocturnal use, has little inconvenience, because ventilation during the night often suffices. Major advantages include correction of hypoventilation during autonomous breathing time that is usually sufficient to permit patients to resume their activities of daily living without need for ventilatory assistance during the day and efficacy comparable to that of intermittent positive pressure ventilation via an indwelling tracheostomy tube, without the inconveniences (tracheostomy is always available if necessary). PMID- 9390882 TI - Chronic noninvasive ventilation in obstructive airways disease. AB - Early experience with noninvasive ventilation in patients with chronic obstructive pulmonary disease was largely unsuccessful, with no benefit over long term oxygen therapy. Nasal positive pressure ventilation, however, produces improvements in arterial blood gases, nocturnal-hypoventilation, and quality of life. Experience with nasal ventilation in other obstructive airways diseases, such as bronchiectasis and cystic fibrosis, is limited and less favorable. PMID- 9390883 TI - Noninvasive ventilation in acute respiratory failure. AB - Noninvasive PPV has been employed for decades in patients with chronic respiratory failure. Increasing use in patients with acute respiratory failure is a more recent phenomenon, mainly because of advances in noninvasive interfaces and ventilator modes. Noninvasive PPV delivered by nasal or oronasal mask has been demonstrated to reduce the need for endotracheal intubation, decrease lengths of stay in the ICU and hospital, and possibly reduce mortality. In the acute care setting, evidence now demonstrates the efficacy of noninvasive PPV for acute exacerbations of COPD, pulmonary edema, pulmonary contusions, and acute respiratory failure in patients who decline or who are not believed to be candidates for intubation. No firm conclusions can yet be made regarding patients with respiratory failure due to other causes, but studies suggest that noninvasive PPV may also be of benefit in patients with postoperative respiratory insufficiency, chest wall disease, and cystic fibrosis. Several factors are vital to the success of this therapy, including careful patient selection, properly timed intervention, a comfortable, well-fitting interface, patient coaching and encouragement, and careful monitoring. Noninvasive ventilation should be used as a way to avoid endotracheal intubation rather than as an alternative. Accordingly, a trial of noninvasive ventilation should be instituted in the course of acute respiratory failure before respiratory arrest is imminent, to provide ventilatory assistance while the factors responsible for the respiratory failure are aggressively treated. Moreover, the authors favor conservative management with expeditious intubation in patients who have other conditions that place them at risk during use of noninvasive ventilation or in patients failing to respond to noninvasive PPV. Noninvasive PPV clearly represents an important addition to the techniques available to manage patients with acute respiratory failure; however, because most studies have been retrospective and uncontrolled, many issues remain unresolved. Further controlled studies are needed to confirm the safety and efficacy of noninvasive PPV, evaluate the most appropriate selection of patients and timing of intervention, define the best type of interface, and assess the costs of noninvasive PPV in comparison with conventional therapy. PMID- 9390884 TI - Noninvasive ventilation for postoperative support and facilitation of weaning. AB - Noninvasive ventilation includes continuous positive airway pressure with mask, positive pressure ventilation with mask, and negative pressure body ventilation. Noninvasive ventilation is a ventilatory support mode intermediate in both effectiveness and potential complications between oxygen administration and intubation with mechanical ventilation. The advantages, disadvantages, and experimental results of the use of noninvasive ventilation in patients who have been extubated following recovery from surgery and subsequently experience respiratory difficulties are discussed. In general, noninvasive ventilation seems to provide useful ventilatory support in about three of four patients in whom it is tried. In addition, the use of noninvasive ventilation as an aid to weaning of patients from mechanical ventilation is discussed. This use of noninvasive ventilation has not yet been extensively reported, although it appears to be potentially useful. PMID- 9390885 TI - Respiratory management, survival, and quality of life for high-level traumatic tetraplegics. AB - Although spinal cord injury is devastating and can compromise the respiratory system, particularly when the cervical cord is injured, aggressive use of noninvasive respiratory muscle aids can reduce the otherwise commonly seen complications of pneumonia, bronchial mucous plugging, atelectasis, and respiratory failure. Accessory muscle function can also usually be improved and the muscles then recruited to help maintain adequate alveolar ventilation during the acute spinal cord injury recovery period. Noninvasive assisted ventilation can be successful for patients with compromised lung function during the acute rehabilitation period as well as on a long-term basis. Removal of an indwelling tracheostomy tube results in improved quality of life from many points of view, a decreased number of local tracheostomy complications, a decreased number of serious respiratory infections, an improved ability to communicate, and an increased ability to use the mouth for functions such as operating computers and wheelchairs. PMID- 9390886 TI - Noninvasive clearance of airway secretions. AB - Airway clearance techniques are indicated for specific diseases that have known clearance abnormalities (Table 2). Murray and others have commented that such techniques are required only for patients with a daily sputum production of greater than 30 mL. The authors have observed that patients with diseases known to cause clearance abnormalities can have sputum clearance with some techniques, such as positive expiratory pressure, autogenic drainage, and active cycle of breathing techniques, when PDPV has not been effective. Hasani et al has shown that use of the forced exhalatory technique in patients with nonproductive cough still resulted in movement of secretions proximally from all regions of the lung in patients with airway obstruction. It is therefore reasonable to consider airway clearance techniques for any patient who has a disease known to alter mucous clearance, including CF, dyskinetic cilia syndromes, and bronchiectasis from any cause. Patients with atelectasis from mucous plugs and hypersecretory states, such as asthma and chronic bronchitis, patients with pain secondary to surgical procedures, and patients with neuromuscular disease, weak cough, and abnormal patency of the airway may also benefit from the application of airway clearance techniques. Infants and children up to 3 years of age with airway clearance problems need to be treated with PDPV. Manual percussion with hands alone or a flexible face mask or cup and small mechanical vibrator/percussors, such as the ultrasonic devices, can be used. The intrapulmonary percussive ventilator shows growing promise in this area. The high-frequency oscillator is not supplied with vests of appropriate sizes for tiny babies and has not been studied in this group. Young patients with neuromuscular disease may require assisted ventilation and airway oscillations can be applied. CPAP alone has been shown to improve achievable flow rates that will increase air-liquid interactions for patients with these diseases or airway malacia. Use of positive pressure to maintain airway patency in these children allows cephalad clearance of secretions. Patients with segmental atelectasis, particularly related to asthma, may benefit from intrapulmonary percussive ventilator, positive expiratory pressure, or PDPV. Prevention of postoperative atelectasis is particularly well suited to positive expiratory pressure, which is not as painful as techniques using oscillations. Neurologically abnormal patients who are unable to cooperate with any active method are also treated using intrapulmonary percussive ventilator, PDPV, and suctioning, if necessary. Musculoskeletal abnormalities, muscular dystrophies, myasthenia gravis, poliomyelitis, or other similar diseases require stabilization of bellows function. Optimizing ventilation in patients with such abnormalities may require positive pressure ventilation either during sleep or continuously. Externally applied pressure, such as with the In Exsufflator or the cyclically inflated pneumatic belt, can augment the patient's own efforts and is sometimes helpful. Normalizing the vital capacity and functional residual capacity typically helps to improve the ability to cough and clear secretions. Assisted cough devices or maneuvers are described in other papers by Bach and Hill. Not all patients who have weak muscles require nocturnal or continuous support, and may benefit from positive expiratory pressure mask treatments. Further studies are sorely needed for this population. Long-term controlled trials are urgently needed to help establish the best types of treatment for patients with CF and bronchiectasis. Such studies will become more complicated by the introduction of new treatments, such as DNase and other therapies that alter secretions, and may begin to change mucociliary or cough clearance. The selection of appropriate outcome measures is central to studying these questions, and it is unclear which are the most important. (ABSTRACT TRUNCATED) PMID- 9390887 TI - Ventilatory support in the field. AB - Ventilatory support during cardiopulmonary resuscitation can be accomplished with an array of methods and devices. These run the gamut from expired air resuscitation, including mouth-to-mouth and mouth-to-mask, to the use of ventilators including ventilator-to-mask and ventilator-to-artificial airway techniques. Appropriate application of these techniques depends on the clinical situation, rescuer training, and availability of equipment. This article discusses the proposed standards of emergency ventilatory support, the advantages and disadvantages of the techniques and devices used, and current controversies surrounding this topic. PMID- 9390888 TI - Respiratory issues in aeromedical patient transport. AB - Respiratory considerations in aeromedically evacuated patients are the cornerstone of safe, successful transport. Maintenance of the ABCs and ongoing resuscitation including pulmonary/ventilator stabilization and management en route are paramount. All of these goals are predicated on a well-developed understanding of hypobaric pulmonary physiology and hypobaric effects on medical devices, a solid grasp of the inherent limits of an aeromedical environment, and the resolute accomplishment of both initial and follow-up team member training. PMID- 9390889 TI - Pulmonary consequences of severe chest trauma. AB - Combined flail chest and pulmonary contusion is a frequent problem in patients with blunt multisystem trauma admitted to the intensive care unit. These patients are at high risk for pneumonia and adult respiratory distress syndrome, which adds substantially to their morbidity and mortality rates. This article discusses the epidemiology and pathophysiology of this condition and the role of the respiratory care practitioner in the optimal management of these critically injured patients. PMID- 9390890 TI - Ventilatory support of the trauma patient with pulmonary contusion. AB - Pulmonary contusion is a common clinical condition seen in patients with thoracic trauma. The mortality rate from isolated pulmonary contusion is low, but when combined with other severe injuries patients with pulmonary contusion can have mortality rates as high as 10% to 57%. Early aggressive mechanical ventilatory support can prevent progressive atelectasis and worsening of arterial oxygenation. Careful titration of ventilatory support to physiologic end points is a reasonable approach. Unfortunately, data comparing the means of achieving the end points are lacking and no published prospective, randomized trials demonstrate superiority of any particular mode of support. Careful attention to detail and physiologic management of the patient in a goal-directed fashion using a multidisciplinary team approach involving the respiratory care practitioner, nurse, and physician is the current state of the art in the management of patients with pulmonary contusion. PMID- 9390891 TI - Management of blunt chest injury. AB - The development of regional techniques of analgesia has revolutionized the management of blunt thoracic trauma. The standard of care has evolved from intubation and mechanical ventilation for all patients to optimization of pain control combined with chest physiotherapy. Although many methods have been used, it appears that in appropriately selected patients, epidural analgesia is the preferred technique for pain control in severe thoracic trauma. Epidural catheters for continuous narcotic or local anesthetic administration are both the most reliable and the most effective, and once in place they can be managed by nursing outside of the intensive care setting. It still remains for improvement in outcome to be demonstrated when epidural analgesia is used, but it is clear that subjective patient comfort is increased and that pulmonary parameters can be improved. In appropriately selected patients, those without head injury or who have been adequately evaluated for intra-abdominal injury, epidural analgesia is currently the preferred method for pain control following severe thoracic trauma. PMID- 9390892 TI - Outcomes of pediatric mechanical ventilation. AB - Mechanical ventilation is one of the most commonly used life-support technologies in the PICU, is an absolute indicator of the need for PICU care, and adds considerably to the cost of intensive care. Patients with chronic disease account for a large proportion of admissions to the PICU and of patients undergoing cardiovascular surgery and neurosurgery. Although the average length of use of mechanical ventilation in the PICU is about 5 days, there is tremendous variability in the length of ventilation (SD approximately 9 days). Survival among ventilated patients varies greatly and depends mostly on the nature and severity of the underlying disease. PMID- 9390893 TI - Pediatric mechanical ventilation technology. AB - An in-depth examination of ventilators currently marketed for the pediatric population and the technology associated with each is provided with this article. Also included is a discussion of an "ideal pediatric ventilator" and its application to the pediatric intensive care patient. Triggering, cycling, and limiting variability, costs, and special features currently available are detailed. PMID- 9390895 TI - High-frequency oscillatory ventilation in pediatric patients. AB - High-frequency oscillatory ventilation (HFOV) offers the potential to maintain adequate gas exchange without imposing the large pressure swings and tidal volumes associated with ventilatory-induced lung injury. This article reviews the studies evaluating the use of HFOV to treat pediatric respiratory failure, discusses the complications associated with HFOV, and details an approach to the practical application of HFOV in the non-neonatal pediatric population. PMID- 9390894 TI - Sedation, analgesia, and neuromuscular blockade during pediatric mechanical ventilation. AB - The mechanically ventilated PICU patient is subjected to multiple noxious stimuli ranging from a bright, noisy, and intimidating environment to painful but necessary procedures. His or her primary disease process or processes obviously constitutes another potential source of noxious stimuli as well. As a result, these patients almost certainly need some combination of medications to allay anxiety, treat discomfort, and perhaps otherwise optimize medical management. Intensivists now have at hand an impressive array of medications that can be used to blunt the stresses imposed by these stimuli. Sedatives to induce anxiolysis and calmness, analgesics to alleviate pain, and occasionally neuromuscular blocking agents to inhibit movement may be used. Use of these medications can be tailored to meet the varied requirements of the diverse PICU population. The consequences of incorrect use can be sobering. Familiarity with the pharmacology, indications, and side effects of the individual medications is a necessity for all ICU care providers to prevent misuse. Although the frequent need for analgesics, sedatives, and NMBDs in the PICU is undisputed, the development of reliable methods for accurately assessing the degree of patient sedation or analgesia will greatly facilitate efforts to improve patient care Appropriate use of sedatives, analgesics, and NMBDs provides an invaluable service. It is important to remember, however, that even in the high-technology PICU environment verbal and physical reassurance remains a powerful tool for providing comfort and anxiolysis to critically ill children. There is no pharmacologic equivalent of human compassion. PMID- 9390896 TI - High-frequency jet ventilation in the pediatric intensive care unit. AB - Scientific data and anecdote on the utility of HFJV in children remain divergent. There seems to be some evidence that ventilation and oxygenation in critically ill patients can sometimes be achieved at lower peak airway pressures and tidal volumes. There is also growing consensus that HFJV may be a superior ventilatory technique for patients with air leak syndrome. However, conclusive evidence of the effect of this technology on overall survival, length of ventilation, or even resource consumption remains illusive. Until such time, a general recommendation for the use of HFJV in pediatric patients is not possible. PMID- 9390897 TI - Metabolic measurements during mechanical ventilation in the pediatric intensive care unit. AB - The metabolism of critically ill infants and children is significantly influenced by their underlying diseases, and standard predictive equations result in inappropriate nutritional support in most of these patients. Furthermore, significant day-to-day variability in energy expenditure may be present in individual patients. Inadequate or excessive energy supply can adversely affect the clinical course of many critically ill patients. For these reasons, serial measurements of energy expenditure should be considered whenever accurate determination of energy needs is deemed necessary. With the wide availability of proprietary metabolic carts suitable for use in mechanically ventilated pediatric patients, serial metabolic measurements via indirect calorimetry are feasible in most critically ill infants and children. The use of indirect calorimetry should also be considered in this population to assess changes in oxygen consumption and the relationship of oxygen consumption to oxygen delivery in response to changes in therapy, such as manipulation of cardiac output using vasoactive medications, or during weaning of mechanical ventilation. PMID- 9390898 TI - Nitric oxide administration during pediatric mechanical ventilation. AB - The administration of inhaled NO in the management of pulmonary hypertension and respiratory failure is an important development in pediatric critical care. The long-term effects of this therapy are yet to be understood. An appreciation for the potential hazards and the development of accurate and safe delivery systems are paramount to the successful application of this new therapy. PMID- 9390899 TI - Volume and pressure modes of mechanical ventilation in pediatric patients. AB - There are few controlled pediatric studies comparing the various modes of ventilation in terms of patient outcomes. Thus at this time the choice of ventilator mode depends largely on the apparatus available, the patient's disease state, and personal preference based on one's experience. The next generation of ventilators may well allow the use of the best of both modes, setting both pressure and volume minimums and maximums, safely meeting ventilation targets. Today's challenges are to become familiar with the various modes of ventilators available, understand the developing physiology of the lung and lung disease pathophysiology, and incorporate all this into proper ventilator strategies to prevent ventilator-induced lung injury. PMID- 9390900 TI - Early versus late tracheostomy in the trauma patient. AB - The use of early tracheostomy in the multiply injured trauma patient has many advantages both in terms of patient management and reduction of morbidity associated with prolonged translaryngeal intubation. Tracheostomy (percutaneous or open technique) has been associated with very low risk of mortality and comparable morbidity to prolonged endotracheal intubation. There exist improved clinical criteria for predicting which patients will require prolonged mechanical ventilation in the trauma and critical care setting. A delay in converting translaryngeal intubation to tracheostomy had been associated with longer ICU stays; conversely, early tracheostomy has been associated with a reduction in ICU stays, incidence of hospital-acquired pneumonias, mechanically ventilated days, and length of hospital stay. Thus, the benefits of early tracheostomy are improved care for patients in the trauma or critical care setting and reduced hospital and patient costs. PMID- 9390901 TI - Ventilatory support following burns and smoke-inhalation injury. AB - The first major improvement in the treatment of burn injury came with the recognition of the importance of fluid resuscitation to prevent shock and renal failure. Subsequently, the use of topical antibiotics to control burn-wound infection and prevent invasive burn-wound sepsis led to the next significant reduction in morbidity and mortality of burn patients. Although progress has been made in the treatment of inhalation injury, the pathophysiology of the injury is still incompletely defined. A better understanding of pathogenic mechanisms will lead to the development of therapeutic agents and treatment regimens that will modulate the cascades of humoral mediators of organ dysfunction and reduce the morbidity and mortality associated with inhalation injury. The recognition of ventilator-induced lung injury has led to adoption of alternative ventilatory techniques such as high-frequency percussive ventilation, which has been shown to substantially reduce the morbidity associated with inhalation injury. PMID- 9390902 TI - Respiratory support of the head-injured patient. AB - Approximately 500,000 patients present with a CHI annually in the United States alone. Up to 20% of these injuries are classified as severe. Appropriate and aggressive intensive care of CHI patients will certainly reduce both the morbidity and mortality rates. Early therapy includes provision of adequate ventilation and oxygenation and definitive care based on clinical assessment. Once the acute phase of the injury has passed, supportive therapy should be maintained as long as secondary injury or complications are avoided. Respiratory care of CHI patients is important though different in each phase of the disease. Proper placement of and maintenance of airways, efficient use of and withdrawal from the mechanical ventilator, and providing adequate pulmonary toilet in order to treat or avoid pneumonia are but a few of the very important respiratory care practices necessary to provide optimal outcomes in patients with CHI. PMID- 9390903 TI - Nutritional support of the mechanically ventilated patient. AB - As with all critically ill patients, those requiring mechanical ventilation are susceptible to the wasting of illness and cannot survive without prompt nutritional support. It may be fair to say that the proper provision of nutrients, and in particular the avoidance of overfeeding, are even more crucial for this subset of critically ill patients. To maximize the overall benefits of feeding, it is crucial to provide the nutritional support early and enterally whenever possible. Therefore, the best strategy for early removal of the mechanical ventilatory support must include the timely and careful administration of nutrients, micronutrients, minerals, vitamins, and fluid, in conjunction with standard intensive care therapeutics and the appropriate respiratory muscle strengthening program. PMID- 9390904 TI - Paralyzation and sedation of the ventilated patient. AB - The selection of pharmacologic agents for the sedation and paralysis of critically ill patients should be based on clinical and pharmacoeconomic trials in this patient population. There is a need for the design and evaluation of cost effective regimens, especially with the continued development and release of newer agents. Additionally, clinicians must continue to be sensitive to the monitoring techniques of sedation and paralysis to ensure maximal clinical benefit while minimizing the adverse effect profile of pharmacologic agents. PMID- 9390905 TI - To everything turn, turn, turn.... An overview of continuous lateral rotational therapy. AB - Continuous lateral rotational therapy can be a significant adjunct in the care of the critically ill patient. CLRT has a great impact on both patient outcomes as well as cost containment in the care of the critically ill. These systems should be used with clear guidelines to determine when CLRT is indicated, its therapeutic benefit, and when to discontinue the therapy. Much research is needed to validate efficacy of particular systems, cost-effectiveness, and necessary frequency and degree of rotation to attain optimal clinical benefits. PMID- 9390906 TI - Integrating education with diagnostics. Patient and technologist. AB - Crape states that in relation to the results of spirometry in the Lung Health Study that perhaps the most important message is that pulmonary function technologists need continuous monitoring and feedback to maintain optimum performance. Technologist education is an essential component of producing valid test results. PMID- 9390907 TI - Spirometry. AB - Spirometry is the most widely used pulmonary function test. It is used for diagnosis and monitoring in a wide variety of obstructive and restrictive disease patterns. Spirometry is the primary measure in determining disability due to pulmonary disease, and is widely applied in the evaluation of bronchodilator response and airway hyperreactivity. A wide variety of spirometry equipment is currently available, from small portable units to large laboratory systems capable of multiple functions. Most spirometers rely heavily on computerization, which makes them easy to use, but require training and experience on the part of the user. Valid spirometry demands equipment that conforms to recommended standards, and performance of the tests to meet criteria for acceptability and reproducibility. Interpretation of spirometry requires attention to these standards and to careful selection of reference values. PMID- 9390908 TI - Derivation, application, and utility of static lung volume measurements. AB - Measurement of static lung volumes remains an important tool to verify and quantify the existence of pulmonary dysfunction. The utility of these measurements, however, is dependent on the knowledge and skills of the technologist, who must ensure that the equipment is operating in an acceptable fashion as well as ensuring that the subject is performing the tests as directed. The technologist as well as the physician interpreting the final results must be cognizant of the limitations associated with each procedure employed in the testing process and be able to verify if the derived data are compatible with the patient's clinical presentation. Laboratories are encouraged to develop their own quality-control programs and monitors to ensure testing is in accordance with published recommendations. By standardizing the testing process, variability is reduced. Ultimately, test quality and patient management will improve. PMID- 9390909 TI - Diffusing capacity of the lung for carbon monoxide. AB - The measurement of co uptake (VCO and DLCO) from alveolar gas is a unique way to noninvasively assess pulmonary vascular function, specifically the functional volume of the pulmonary capillary bed. Proper interpretation of results, however, needs to account for inherent assumptions regarding co distribution and timing procedures. Moreover, reasonable airway mechanics, lung volumes, and patient cooperation are required for accurate measurements. Potential clinical utility may be increased if measurements are made in different positions or under exercise conditions. PMID- 9390910 TI - Airway resistance measurements. AB - Resistance measurements are the most useful parameters for assessing acute changes in airway caliber associated with bronchodilation or bronchial provocation. Used in addition to spirometry, Raw can provide a better differentiation of the causes of airflow impairment as well as the presence of concurrent processes. A simple, noninvasive Raw measurement can provide definitive answers in the absence of other changes. Finally, the addition of practical, nonplethysmographic measurements opens a new application for bedside, office, and home monitoring. PMID- 9390911 TI - Exercise testing. PMID- 9390912 TI - Quality assurance. AB - Every pulmonary function laboratory should develop and implement a quality assurance program to minimize various technical sources of variation. This article has discussed six major components. First, the education and training of the technologists in the pulmonary function laboratory is probably the most important factor in obtaining accurate and reproducible results. A college-level education with an emphasis on math and science is recommended. After an appropriate training program, continued evaluation and feedback are important. Second, instrument maintenance should be performed on a scheduled basis to reduce or prevent instrument malfunctions. Corrective maintenance, which is usually unscheduled, should be performed by knowledgeable individuals and any repairs should be documented. Third, a procedure manual is very important to any successful quality assurance program. It should contain a broad range of information including administrative issues, quality-control procedures, stepwise instructions on test performance, and infection-control policies and procedures. Fourth, the procedures should be performed using published guidelines to help minimize the effects of the many variables. Fifth, a method to quality control each test procedure should be developed. The specific method(s) will vary according to the type of instrumentation and the manufacturer. Finally, the well run quality assurance program must properly analyze and store the data collected. Sound statistical methods should be applied and various logs and lists should be developed. PMID- 9390913 TI - Indirect calorimetry. AB - Indirect calorimetry can be a useful tool to define nutritional status, determine nutritional requirements, and assess response to nutritional interventions. Measurements of oxygen consumption and carbon dioxide production may be used to determine cardiac output and work of breathing, and estimate the components of minute ventilation. An understanding of the potential technical and physiological pitfalls is necessary to obtain meaningful and useful information. PMID- 9390914 TI - Pulmonary function in the mechanically ventilated patient. AB - Implementing the knowledge and skills obtained from the pulmonary function laboratory is useful in the assessment of pulmonary function in the ventilated patient. In the critically ill patient, special constraints and problems (such as safety and ability to cooperate) are important considerations for the therapist performing the testing. Basic measurements such as MIP, VC, VT, MVV, VE, and respiratory rate are commonly made to assess weanability. For the difficult-to wean patient, WOB, PTP, P0.1 and O2COB may be of potential value in selected patients. Evidence of their value in routine weaning assessment, however, is lacking. Monitoring has evolved with increased understanding of pulmonary mechanics of the mechanically ventilated patient. Measurements of compliance, resistance, MAP, and autoPEEP provide useful diagnostic and therapeutic information, and guide in the selection of appropriate machine settings to provide effective and safe ventilatory support. Although currently confined to the research setting, P-V curves have contributed to our understanding of the pathophysiology of ALI, and may prove useful in defining an optimal ventilatory strategy in patients with ALI. Finally, waveform monitoring can reveal, graphically and in real time, patient-ventilator interactions such as autoPEEP, lung overdistension, patient effort, or the presence of secretions. Waveform monitoring can bring important issues to the close attention of the practitioners involved in management of the ventilated patient. PMID- 9390915 TI - Neonatal pulmonary function testing. AB - Recent advances in technology have made it possible for use to measure neonatal pulmonary function, even in very small infants. Applying PM measurement as a tool for ventilator-patient management improves outcome in the NICU. With PFT in the laboratory, we can diagnose and determine severity of pulmonary illness and evaluate therapies through infancy and childhood. Although the task of safely measuring pulmonary function in infants can be daunting, there is no reason why more PFT labs should not offer services to these patients. PMID- 9390916 TI - The biologic basis for inhaled nitric oxide. AB - Nitric oxide is produced by nitric oxide synthase enzymes, which cleave the amino acid L-arginine to form nitric oxide and the amino acid L-citrulline. Many of the biologic actions of nitric oxide occur because nitric oxide activates guanylate cyclase, which in turn synthesizes a second-messenger molecule, cyclic guanosine 3',5'-monophosphate (cGMP). The increased concentration of cGMP activates cGMP dependent protein kinase, reducing intracellular concentrations of calcium and relaxing smooth muscle. Nitric oxide also has many important effects that may not be mediated through increases of pulmonary cGMP activity. These include the ability to scavenge oxygen free radicals, reduce oxygen toxicity, and inhibit platelet and leukocyte aggregation. Nitric oxide is metabolized and excreted via a number of diverse pathways that may modify the toxicity of the molecule. PMID- 9390917 TI - Delivery systems for inhaled nitric oxide. AB - From a practical standpoint, technical issues related to NO delivery are as important as therapeutic issues. The benefits can be appreciated only if a reliable delivery system is used. Further, hazards and toxicity may be more problematic with an unreliable delivery system. It is incumbent on clinicians using inhaled NO to ensure that the delivery system is safe and reliable. PMID- 9390918 TI - Manufacture and measurement of nitrogen oxides. AB - A decade of research has identified nitric oxide as a unique endogenous biologic mediator with functions as diverse as vasodilation, macrophage cytotoxicity, platelet adhesion, and memory formation. Measurement devices, previously used in research on atmospheric nitrogen oxides, are being adapted for and applied to clinical situations without approval, regulation, or rigorous testing, and with little understanding of how the devices work or their limitations. This article discusses the chemistry, manufacture, and measurement of nitrogen oxides. PMID- 9390919 TI - Use of inhaled nitric oxide for ARDS. AB - Inhalation of low inspired concentration of nitric oxide reduces pulmonary hypertension and increases arterial oxygen tension in patients with acute respiratory distress syndrome (ARDS), and appears to be safe. Research on the physiologic mechanisms regulating the action of inhaled nitric oxide may provide clinicians with ways to further potentiate and prolong its beneficial effects. PMID- 9390920 TI - Inhaled nitric oxide in the management of cardiopulmonary disorders in infants and children. AB - The administration of NO has become an important and effective therapy in the clinical management of pulmonary hypertension associated with cardiopulmonary disorders in infants and children. It is likely to become a routine therapy in the treatment of PPHN, although dosing and timing strategies, early indicators of treatment failure, and long-term outcomes are not completely understood. The use of NO has also been beneficial in the evaluation and management of pulmonary hypertension associated with congenital heart disease. The role of NO in the management of pediatric ARDS holds promise, although further clinical trials are needed. Additional research should also be directed toward the use of NO in preterm infants and those born with CDH. Future endeavors may also include the use of NO in the evaluation and management of asthma. PMID- 9390921 TI - A symposium on controversies in the pathology of transitional cell carcinomas of the urinary bladder. Part I. PMID- 9390922 TI - Flow cytometry phenotyping and molecular techniques in nongynecologic cytology. PMID- 9390923 TI - Mammary lymphoid tissue: a unique component of the mucosal immune system. PMID- 9390924 TI - Temporal artery biopsy diagnosis of giant cell arteritis: lessons from 1109 biopsies. PMID- 9390925 TI - Large needle biopsy of the thyroid gland. PMID- 9390926 TI - Cervical squamous cell carcinoma and its precursor lesions: cytodiagnostic criteria and pitfalls. PMID- 9390927 TI - "God's first cancer and man's first cure": milestones in gestational trophoblastic disease. PMID- 9390928 TI - Transitional cell carcinoma and other transitional cell tumors of the ovary. PMID- 9390929 TI - An epidemiologic study of differences between mammary ductal and lobular carcinoma in situ: analysis of 11,436 cases from the California Cancer Registry. PMID- 9390930 TI - Multiple papillomas of the breast: morphologic findings and clinical evolution. PMID- 9390931 TI - Latitudinal variation in the abundance and oxidative capacities of muscle mitochondria in perciform fishes AB - The abundance, distribution and oxidative capacities of mitochondria have been investigated in the red pectoral fin adductor muscles of fish (Order Perciformes) that use a predominantly labriform style of swimming. Mediterranean Sea species from the families Labridae, Serranidae, Sparidae and Antarctic Nototheniidae and non-Antarctic Nototheniidae and Channichthyidae were studied. Sub-Antarctic species from the Beagle Channel, Tierra del Fuego, included the pelagic haemoglobin-less icefish (Champsocephalus esox) and the robalo (Eleginops maclovinus), which occurs as far north as 35 degrees S. In Champsocephalus esox, the mitochondrial volume density of red muscle was 0.51 and mitochondrial cristae surface density (43. 9 microm2 microm-3) was higher than reported for Antarctic icefishes. In the red-blooded, active pelagic or semi-pelagic species, mitochondrial volume density was within the range 0.27-0.33 regardless of habitat temperature. Amongst less active demersal species, mitochondrial volume density ranged from 0.29-0.33 in polar species to 0.08-0.13 in Mediterranean species. In Antarctic species and Champsocephalus esox, myofibrils occurred in ribbons or clusters one fibril thick entirely surrounded by mitochondria. The volume density of intracellular lipid droplets was not correlated with activity patterns or habitat temperature. In a comparison of Eleginops maclovinus caught in summer (approximately 10 degrees C) and winter (approximately 4 degrees C), mitochondrial volume density did not differ, whereas the surface density of mitochondrial clusters was higher in summer fish. The temperature-dependence of the state 3 respiration rate of isolated mitochondria with pyruvate as substrate was described by a single quadratic relationship for all species, indicating no significant up-regulation of the maximum rate of oxygen uptake per milligram mitochondrial protein in Antarctic species. Our results support the conclusion that increasing the volume and surface density of mitochondrial clusters is the primary mechanism for enhancing the aerobic capacity of muscle in cold-water fish. PMID- 9390932 TI - Membrane potential responses of paramecium caudatum to bitter substances: existence of multiple pathways for bitter responses AB - The membrane potential responses of Paramecium caudatum to the external application of bitter substances were examined by employing conventional electrophysiological techniques. Mutant cells defective in voltage-gated Ca2+ channels were used to record the potential responses in the absence of contamination by Ca2+ action potentials. The cells produced a transient depolarization followed by a transient hyperpolarization in response to a rapid whole-cell application of chloroquine, strychnine nitrate or brucine. Of these chemicals, chloroquine was the most potent. Cells produced a simple depolarization in response to a localized application of test chemicals to the anterior region, whereas they produced a transient hyperpolarization in response to an application to the posterior region. Membrane potential responses to an application of chloroquine declined with repeated application. The presence of chloroquine in the external bathing solution strongly inhibited the membrane potential responses to an application of brucine or strychnine. However, the presence of chloroquine did not affect the membrane potential responses to an application of quinine. It is suggested that chloroquine, strychnine and brucine share a common component of their transduction pathways, but that the transduction pathway for quinine is different. PMID- 9390933 TI - Alterations of ionic membrane permeabilities in multidrug-resistant neuroblastoma x glioma hybrid cells. AB - A population of NG108-15 neuroblastoma cells resistant to doxorubicin (NG/DOXR) was established. The cells exhibited a multidrug resistance phenotype with cross resistance to vinblastin and colchicine, overexpression of a 170 kDa membrane protein identified as P-glycoprotein and reversal of resistance by verapamil and quinine. Compared with NG108-15 cells, NG/DOXR cells showed an increase in Na+ current density and a decrease in cyclic-AMP-activated Cl- current density with no change in K+- and volume-sensitive Cl- current densities. As previously observed in NG108-15 cells, the vacuolar-type H+-ATPase inhibitors bafilomycin A1 and nitrate induced membrane depolarizations in NG/DOXR cells. The resting potentials of sensitive and resistant cells were not significantly different, but the depolarizations evoked by these agents were significantly larger in NG/DOXR than in NG108-15 cells. The resting membrane potential of NG/DOXR cells, but not that of NG108-15 cells, was depolarized by verapamil, and this effect was abolished by bafilomycin. The volume-sensitive Cl- currents of drug-sensitive and drug-resistant cells were inhibited by a decrease in intracellular pH from 7.3 to 6.8. Whereas bafilomycin prevents activation of Cl- currents in both drug sensitive and drug-resistant cells, verapamil inhibited the Cl- current only in NG/DOXR cells. The results are discussed in terms of the roles of cytoplasmic pH and membrane potential in multidrug resistance. PMID- 9390935 TI - Acid-base regulation, metabolism and energetics in sipunculus nudus as a function of ambient carbon dioxide level AB - Changes in the rates of oxygen consumption and ammonium excretion, in intra- and extracellular acid-base status and in the rate of H+-equivalent ion transfer between animals and ambient water were measured during environmental hypercapnia in the peanut worm Sipunculus nudus. During exposure to 1 % CO2 in air, intracellular and coelomic plasma PCO2 values rose to levels above those expected from the increase in ambient CO2 tension. Simultaneously, coelomic plasma PO2 was reduced below control values. The rise in PCO2 also induced a fall in intra- and extracellular pH, but intracellular pH was rapidly and completely restored. This was achieved during the early period of hypercapnia at the expense of a non respiratory increase in the extracellular acidosis. The pH of the extracellular space was only partially compensated (by 37 %) during long-term hypercapnia. The net release of basic equivalents under control conditions turned to a net release of protons to the ambient water before a net, albeit reduced, rate of base release was re-established after a new steady state had been achieved with respect to acid-base parameters. Hypercapnia also affected the mode and rate of metabolism. It caused the rate of oxygen consumption to fall, whereas the rate of ammonium excretion remained constant or even increased, reflecting a reduction of the O/N ratio in both cases. The transient intracellular acidosis preceded a depletion of the phosphagen phospho-l-arginine, an accumulation of free ADP and a decrease in the level of Gibbs free energy change of ATP hydrolysis, before replenishment of phosphagen and restoration of pHi and energy status occurred in parallel. In conclusion, long-term hypercapnia in vivo causes metabolic depression, a parallel shift in acid-base status and increased gas partial pressure gradients, which are related to a reduction in ventilatory activity. The steady-state rise in H+-equivalent ion transfer to the environment reflects an increased rate of production of protons by metabolism. This observation and the reduction of the O/N ratio suggest that a shift to protein/amino acid catabolism has taken place. Metabolic depression prevails, with completely compensated intracellular acidosis during long-term hypercapnia eliminating intracellular pH as a significant factor in the regulation of metabolic rate in vivo. Fluctuating levels of the phosphagen, of free ADP and in the ATP free energy change values independent of pH are interpreted as being correlated with oscillating ATP turnover rates during early hypercapnia and as reflecting a tight coupling of ATP turnover and energy status via the level of free ADP. PMID- 9390934 TI - Intergeneric distribution and immunolocalization of a putative odorant-binding protein in true bugs (Hemiptera, Heteroptera). AB - Lygus antennal protein (LAP) is an olfactory-related protein of the tarnished plant bug Lygus lineolaris (Hemiptera, Heteroptera: Miridae), a hemimetabolous insect. In previous work, a polyclonal antiserum was generated against the N terminal sequence of LAP; LAP immunoreactivity was strongest in antennae of adult males, but was also present in antennae of adult females and of nymphs. In the current study, LAP immunoreactivity was examined to determine the species specificity and the tissue and cellular localization of LAP expression. Western blot analysis indicated that LAP immunoreactivity was present in the antennae of the male congeners L. lineolaris and L. hesperous, but was not detectable in male antennae of the more distant relatives Podisus maculiventris or Nezara viridula (Hemiptera, Heteroptera: Pentatomidae). Western blot analysis further confirmed that LAP expression was restricted to antennal tissue. Histological analyses showed that LAP expression within the antennae was specifically associated with chemosensory sensilla on the antenna. Within the sensilla, LAP immunoreactivity was distributed throughout the extracellular lumen and was concentrated in dense granules within the cytoplasm of sensillar support cells. LAP immunoreactivity was restricted to a subset of antennal chemosensory sensilla, specifically the multiporous olfactory sensilla. These findings suggest that LAP has an important olfactory function in Lygus sp., possibly related to that of odorant-binding proteins (OBP) found in other insect orders. If so, LAP would be the first OBP like protein characterized outside the Endopterygota. PMID- 9390936 TI - BCECF in single cultured cells: inhomogeneous distribution but homogeneous response. AB - Using confocal laser scanning microscopy with a dual-wavelength laser system, the behaviour of BCECF [(2',7'-bis-2-carboxyethyl)-5-(and-6)carboxyfluorescein] was investigated in a variety of cell lines. Selection of a small area for monitoring allowed discrimination between various intracellular organelles, whose identity was established by vital staining. It was found that, after loading the cells with BCECF, both the nucleus and the mitochondria showed a higher level of fluorescence than the cytoplasm. Calibration of the pH-sensitivity of these fluorescence signals using the nigericin method yielded identical curves, as did exposure of the cells to NH4Cl. These studies suggest that BCECF, despite its inhomogeneous intracellular distribution, reports the pH of only one cellular compartment, the cytosol. PMID- 9390937 TI - Stresses in human leg muscles in running and jumping determined by force plate analysis and from published magnetic resonance images. AB - Calculation of the stresses exerted by human muscles requires knowledge of their physiological cross-sectional area (PCSA). Magnetic resonance imaging (MRI) has made it possible to measure PCSAs of leg muscles of healthy human subjects, which are much larger than the PCSAs of cadaveric leg muscles that have been used in previous studies. We have used published MRI data, together with our own force plate records and films of running and jumping humans, to calculate stresses in the major groups of leg muscles. Peak stresses in the triceps surae ranged from 100 kN m-2 during take off for standing high jumps to 150 kN m-2 during running at 4 m s-1. In the quadriceps, peak stresses ranged from 190 kN m-2 during standing long jumps to 280 kN m-2 during standing high jumps. Similar stresses were calculated from published measurements of joint moments. These stresses are lower than those previously calculated from cadaveric data, but are in the range expected from physiological experiments on isolated muscles. PMID- 9390938 TI - Effects of temperature on cuticular lipids and water balance in a desert Drosophila: is thermal acclimation beneficial? AB - The desert fruit fly Drosophila mojavensis experiences environmental conditions of high temperature and low humidity. To understand the physiological mechanisms allowing these small insects to survive in such stressful conditions, we studied the effects of thermal acclimation on cuticular lipids and rates of water loss of adult D. mojavensis. Mean hydrocarbon chain length increased at higher temperatures, but cuticular lipid melting temperature (Tm) did not. Lipid quantity doubled in the first 14 days of adult life, but was unaffected by acclimation temperature. Despite these changes in cuticular properties, organismal rates of water loss were unaffected by either acclimation temperature or age. Owing to the smaller body size of warm-acclimated flies, D. mojavensis reared for 14 days at 33 degrees C lost water more rapidly on a mass-specific basis than flies acclimated to 25 degrees C or 17 degrees C. Thus, apparently adaptive changes in cuticular lipids do not necessarily result in reduced rates of water loss. Avoidance of high temperatures and desiccating conditions is more likely to contribute to survival in nature than changes in water balance mediated by surface lipids. PMID- 9390940 TI - Peripheral encoding of moving sources by the lateral line system of a sit-and wait predator AB - Video-tape recordings of prey-capture behaviour were made to demonstrate that stargazers can detect and capture prey in the dark and to determine the range of prey movement velocities that resulted in prey capture. Electrophysiological recording techniques were then used to determine how an artificial source (a sphere), moving at speeds within the range of recorded prey movement velocities, was encoded by anterior lateral line nerve fibres innervating the preopercular mandibular canals on the head. A vibrating sphere was also used to measure frequency-response characteristics to determine the bandwidth of response and fibre origin (type of neuromast and location). In order to measure the relevant stimulus parameters likely to govern neural responses, the pressure-gradient pattern produced by the moving sphere was characterised with a pair of miniature hydrophones separated by approximately the same distance as head lateral line canal pores on stargazers. At least four different features of neural response patterns, including direction-dependent changes in the overall envelope of the firing rate pattern, could be predicted on the basis of measured pressure gradient patterns. The dominant features of both the pressure-gradient and neural response patterns were produced by the wake behind the moving sphere, but behavioural observations indicated that stargazers were responding to the bow of an approaching prey, rather than its wake. Although the form of the wake behind the moving sphere is unlikely to be a good match for the stimulus mediating prey detection, these results clearly establish that pressure-gradient patterns are good predictors of neural response patterns. Thus, similar measurements of pressure-gradient patterns produced by more biologically relevant sources can be used to predict peripheral lateral line responses and stimulus features likely to be of key importance. PMID- 9390939 TI - Fatty acids from the cyanobacterium Microcystis aeruginosa with potent inhibitory effects on fish gill Na+/K+-ATPase activity. AB - Fatty acids from two strains of the cyanobacterium Microcystis aeruginosa, PCC 7820 (a strain that produces the hepatotoxin microcystin-LR, MC-LR) and CYA 43 (a strain that produces only small quantities of MC-LR), were extracted, partially characterised and tested for their inhibitory effect on the K+-dependent p nitrophenol phosphatase (pNPPase) activity of tilapia (Oreochromis mossambicus) gill basolateral membrane. Thin-layer chromatography of the lipids from dichloromethane:methanol extracts of M. aeruginosa PCC 7820 and CYA 43, using diethylether:isopropanol:formic acid (100:4.5:2.5) as solvent, yielded five inhibitory products from M. aeruginosa 7820 and six from M. aeruginosa CYA 43. None of these products could be related to MC-LR. The inhibitory behaviour of the products mimics that of a slow, tight-binding inhibitor. The inhibitory activity is removed by incubation of extracts with fatty-acid-free bovine serum albumin (FAF-BSA). However, FAF-BSA only partially reversed the inhibition of K+ dependent pNPPase on fish gills pre-exposed to the extracted products. We conclude that M. aeruginosa strains PCC 7820 and CYA 43 produce fatty acids with potent inhibitory effects on K+-dependent pNPPase. The release of these products following lysis of cyanobacterial blooms may help to explain fish kills through a disturbance of gill functioning. PMID- 9390941 TI - Predictions of the time course of force and power output by dogfish white muscle fibres during brief tetani. AB - The aim of this study was to identify the principal factors that determine the time course of force and power output by muscle during patterns of stimulation and movement similar to those during fish swimming. Fully activated, white muscle fibres isolated from dogfish Scyliorhinus canicula were used to characterize the force-velocity relationship of the contractile component (CC) and the stress strain relationship of the passive, elastic component (SEC) in series with the CC. A simple model of the time course of crossbridge activation during brief contractions was devised. Using the mechanical properties of the CC and SEC and the activation time course, force and power were predicted for brief contractions with constant-velocity movement and also for brief contractions starting at various times during sinusoidal movement. The predicted force and power were compared with observations for these patterns of stimulation and movement. The predictions matched the observations well for the period during stimulation. Matching of force was much less good for some specific conditions during relaxation, the period during which force persists after the end of stimulation. If either the slow rise of activation or the SEC was omitted from the calculation, the predictions were poor, even during stimulation. Additional factors which may influence force are discussed. These include the after-effects of shortening and stretch, the variation of force during constant-velocity stretch and non-uniform behaviour within the muscle. PMID- 9390943 TI - Faecal firing in a skipper caterpillar is pressure-driven AB - Many leaf-rolling caterpillars have a rigid anal comb attached to the lower surface of the anal plate (or shield) situated above the anus. This comb is widely assumed to be a lever used to 'flick' away frass pellets. An alternative mechanism to explain pellet discharge is proposed on the basis of observations on the caterpillar of the skipper Calpodes ethlius. The model proposes that the underside of the anal plate serves as a blood-pressure-driven surface for the ejection of faecal pellets. Rather than acting as a lever, the anal comb serves as a latch to prevent the premature distortion of the lower wall of the anal plate until the anal haemocoel compartment is fully pressurized. The anal comb is swung into position during pellet extrusion by retractor muscles attached at its base and held in place by a catch formed by a blood-swollen torus of everted rectal wall. As the caterpillar raises the blood pressure in its anal compartment by contracting its anal prolegs, the comb eventually slips over the toral catch. This causes the underside of the anal plate to move rapidly backwards as the blood pressure is released, projecting the pellet resting against it through the air. Simulation suggests that a local blood pressure of at least 10 kPa (75 mmHg) would be required to accelerate the lower surface of the anal plate outwards at a rate fast enough to discharge a 10 mg pellet at an observed mean velocity of 1.3 m s-1. PMID- 9390942 TI - Metal ions suppress the abnormal taste behavior of the Drosophila mutant malvolio. AB - A mutation in the malvolio (mvl) gene affects taste behavior in Drosophila melanogaster. The malvolio gene encodes a protein (MVL) that exhibits homology to the mammalian natural resistance-associated macrophage proteins. It is also homologous to the Smf1 protein from Saccharomyces cerevisiae, which we have recently demonstrated to function as a Mn2+/Zn2+ transporter. We proposed that the Drosophila and mammalian proteins, like the yeast SMF1 gene product, are metal-ion transporters. To test this hypothesis, malvolio mutant flies were allowed to develop, from egg to adulthood, on a medium containing elevated concentrations of metal ions. Mutant flies that were reared in the presence of 10 mmol l-1 MnCl2 or FeCl2 developed into adults with recovered taste behavior. CaCl2 or MgCl2 had no effect on the mutant's taste perception. ZnCl2 inhibited the effect of MnCl2 when both ions were supplied together. Similar suppression of the abnormal taste behavior was observed when mvl mutants were fed MnCl2 or FeCl2 only at the adult stage. Furthermore, exposure of adult mutant flies to these ions in the testing plate for only 2 h was sufficient to restore normal taste behavior. The suppression of the defective taste behavior suggests that MVL functions as a Mn2+/Fe2+ transporter and that Mn2+ and/or Fe2+ are involved in the signal transduction of taste perception in Drosophila adults. PMID- 9390944 TI - Collagen orientation and molecular spacing during creep and stress-relaxation in soft connective tissues. AB - Collagen fibres form cross-helical, cross-ply or quasi-random feltworks in extensible connective tissues; strain-induced reorientation of these networks gives rise to the non-linear mechanical properties of connective tissue at finite strains. Such tissues are also generally viscoelastic (i.e. display time dependent properties). The hypothesis that time-dependent reorientation of collagen fibres is responsible for the viscoelasticity of such tissues is examined here using time-resolved X-ray diffraction measurements during stress relaxation and creep transients applied to rat skin and bovine intramuscular connective tissue. Differences in the intensity and angular orientation of the third and fifth orders of the 67 nm meridional D-spacing of collagen molecules were shown before and after the application of loads or displacements. However, no changes in the D-spacing or angular orientation of collagen occurred during the time course of either stress-relaxation or creep in both tissues. This indicates that collagen fibre reorientation is not a primary source of their viscoelastic properties. The non-linear (strain-dependent) nature of the stress relaxation response in these tissues suggests that relaxation processes within the collagen fibres or at the fibre-matrix interface may be responsible for their viscoelastic nature. PMID- 9390945 TI - Hearing and hunting in red bats (Lasiurus borealis, Vespertilionidae): audiogram and ear properties. AB - We examined aspects of hearing in the red bat (Lasiurus borealis) related to its use of biosonar. Evoked potential audiograms, obtained from volume-conducted auditory brainstem responses, were obtained in two bats, and the sound pressure transformation of the pinna was measured in three specimens. Field-recorded echolocation signals were analysed for comparison. The fundamental sonar search calls sweep from 45 to 30 kHz (peak energy at 35 kHz), approach-phase calls sweep from 65 to 35 kHz (peak 40 kHz) and terminal calls sweep from 70 to 30 kHz (peak 45 kHz). The most sensitive region of the audiogram extended from 10 kHz to 45-55 kHz, with maximum sensitivity as low as 20 dB SPL occurring between 25 and 30 kHz. A relative threshold minimum occurred between 40 and 50 kHz. With increasing frequency, the acoustic axis of the pinna moves upwards and medially. The sound pressure transformation was noteworthy near 40-45 kHz; the acoustic axis was closest to the midline, the -3 dB acceptance angles showed local minima, and the pinna gain and interaural intensity difference were maximal. These results are related to the known echolocation and foraging behavior of this species and match the spectral components of approach- and final-phase calls. We conclude that co evolution with hearing prey has put a higher selective pressure on optimizing localization and tracking of prey than on improving detection performance. PMID- 9390946 TI - Representation of behaviorally relevant sound frequencies by auditory receptors in the cricket teleogryllus oceanicus AB - Teleogryllus oceanicus is particularly sensitive to two ranges of sound frequency, one corresponding to intraspecific acoustical signals (4-5 kHz) and the other to the echolocation cries of bats (25-50 kHz). We recorded summed responses of the auditory nerve to stimuli in these two ranges. Nerve responses consist of trains of compound action potentials (CAPs), each produced by the summed activity of a number of receptor neurons. The amplitude of the CAP is up to four times larger for stimuli at 4.5 kHz than for stimuli at 30 kHz, suggesting either that the extracellular spikes produced by receptors that respond to 4.5 kHz are larger than those that respond to 30 kHz, or that receptors fire more synchronously in response to stimulation at 4.5 kHz, or that more receptors respond to stimulation at 4.5 kHz. Neither unit spike amplitude nor conduction velocity (which is expected to vary with spike amplitude) differs for the two frequencies, and the responses to 4.5 kHz are not produced by more tightly synchronized receptor populations, as judged by CAP breadth. We conclude that more receptors respond to 4. 5 kHz than to 30 kHz. PMID- 9390947 TI - Nitric oxide and vasodilation in human limbs. AB - Both the skeletal muscle and skin of humans possess remarkable abilities to vasodilate. Marked vasodilation can be seen in these vascular beds in response to a variety of common physiological stimuli. These stimuli include reactive hyperemia (skin and muscle), exercise hyperemia (muscle), mental stress (muscle), and whole body heating (skin). The physiological mechanisms that cause vasodilation in response to these stimuli are poorly understood, and the substance(s) responsible for it remain unclear. In this context, recent attention has been focused on the possible contribution of nitric oxide (NO) to the regulation of hyperemic responses in human skin and skeletal muscle. The emerging picture is that NO is not an essential component of the dilator response seen during reactive hyperemia. However, it does appear that NO may play a modest role in exercise hyperemia. NO appears to play a major role in the skeletal muscle vasodilation seen in response to mental stress in humans. Preliminary evidence also indicates that NO is not essential for the normal dilator responses observed in the cutaneous circulation during body heating in humans, but this issue needs further study. There are a number of possible mechanisms that might mediate NO release in humans, and the role of these mechanisms in the various hyperemic responses is also poorly understood. The role of altered NO-mediated vasodilation in some disease states is also discussed. Whereas NO is a potent vasodilating substance, the actions of NO alone do not explain a variety of poorly understood vasodilator mechanisms in conscious humans. Much work remains for those interested in the role of NO in the regulation of blood flow to the skin and skeletal muscle of humans. PMID- 9390948 TI - Invited editorial on "Coupled vs. uncoupled pericardial constraint: effects on cardiac chamber interactions". PMID- 9390949 TI - Coupled vs. uncoupled pericardial constraint: effects on cardiac chamber interactions. AB - The effects of pericardial constraint on cardiac chamber interactions were evaluated by mathematical model analyses based on a novel concept of coupled vs. uncoupled pericardial constraint. We hypothesized that the nature of pericardial constraint can be classified as a "coupled" constraint exerted by uniform liquid pressure or an "uncoupled" constraint exerted by regional surface pressure. The numerical solution of the model of atrioventricular interaction produced the characteristic waveforms in venous flows and right atrial/ventricular pressures in classical pericardial diseases. Coupled constraint accounted for the patterns in cardiac tamponade; uncoupled constraint accounted for those in constrictive pericarditis. Analytic solution of the model of ventricular interdependence demonstrated that coupled constraint (tamponade) produced greater gains in ventricular interdependence, increasing the occurrence of pulsus paradoxus, whereas uncoupled constraint (constriction) produced a greater effective right ventricular elastance, increasing the likelihood of Kussmaul's sign. Thus the concept of coupled vs. uncoupled constraint may offer a coherent framework to understand the characteristic steady-state and respiratory-induced hemodynamic events in multiple forms of pericardial diseases. PMID- 9390950 TI - On the mechanism of mucosal folding in normal and asthmatic airways. AB - Previous studies have demonstrated that the airway wall in asthma and chronic obstructive pulmonary disease is markedly thickened. It has also been observed that when the smooth muscle constricts the mucosa buckles, forming folds that penetrate into the airway lumen. This folding pattern may influence the amount of luminal obstruction associated with smooth muscle activation. A finite-element analysis of a two-layer composite model for an airway is used to investigate the factors that determine the mucosal folding pattern and how it is altered as a result of changes in the thickness or stiffness of the different layers that comprise the airway wall. Results demonstrate that the most critical physical characteristic is the thickness of the thin inner layer of the model. Thickening of this inner layer likely is represented by the enhanced subepithelial collagen deposition seen in asthma. Other findings show a high shear stress at or near the epithelial layer, which may explain the pronounced epithelial sloughing that occurs in asthma, and steep gradients in pressure that could cause significant shifts of liquid between wall compartments or between the wall and luminal or vascular spaces. PMID- 9390951 TI - Hormonal and metabolic responses to exercise across time of day and menstrual cycle phase. AB - Two studies, each utilizing short-term treadmill exercise of a different intensity, assessed the metabolic and hormonal responses of women to exercise in the morning (AM) and late afternoon (PM). In study 1, plasma concentrations of growth hormone, arginine vasopressin, catecholamines, adrenocorticotropic hormone, cortisol, lactate, and glucose were measured before, during, and after high-intensity exercise (90% maximal O2 uptake) in the AM and PM. In study 2, plasma concentrations of adrenocorticotropic hormone, cortisol, lactate, and glucose were measured before, during, and after moderate-intensity exercise (70% maximal O2 uptake) in the AM and PM in the follicular (days 3-9), midcycle (days 10-16), and luteal (days 18-26) phases of the menstrual cycle. The results of studies 1 and 2 revealed no significant diurnal differences in the magnitude of responses for any measured variable. In addition, study 2 revealed a significant time-by-phase interaction for glucose (P = 0. 014). However, net integrated responses were similar across cycle phases. These data suggest that metabolic and hormonal responses to short-term, high-intensity exercise can be assessed with equal reliability in the AM and PM and that there are subtle differences in blood glucose responses to moderate-intensity exercise across menstrual cycle phase. PMID- 9390952 TI - Myocardial blood flow heterogeneity in shock and small-volume resuscitation in pigs with coronary stenosis. AB - Myocardial blood flow heterogeneity in shock and small-volume resuscitation in pigs with coronary stenosis. J. Appl. Physiol. 83(6): 1832-1841, 1997.-We analyzed the effects of shock and small-volume resuscitation in the presence of coronary stenosis on fractal dimension (D) and spatial correlation (SC) of regional myocardial perfusion. Hemorrhagic shock was induced and maintained for 1 h. Pigs were resuscitated with hypertonic saline-dextran 60 [HSDex, 10% of shed blood volume (SBV)] or normal saline (NS; 80% of SBV). Therapy was continued after 30 min with dextran (10% SBV). At baseline, D was 1.39 +/- 0.06 (mean +/- SE; HSDex group) and 1.34 +/- 0.04 (NS group). SC was 0.26 +/- 0.07 (HSDex) and 0.26 +/- 0.04 (NS). Left anterior descending coronary artery stenosis changed neither D nor SC. Shock significantly reduced D (i.e., homogenized perfusion): 1.26 +/- 0.06 (HSDex) and 1.23 +/- 0.05 (NS). SC was increased: 0.41 +/- 0.1 (HSDex) and 0.48 +/- 0.07 (NS). Fluid therapy with HSDex further decreased D to 1.22 +/- 0.05, whereas NS did not change D. SC was increased by both HSDex (0.56 +/- 0.1) and NS (0.53 +/- 0.06). At 1 h after resuscitation, SC was constant in both groups, and D was reduced only in the NS group (1.18 +/- 0.02). We conclude that hemorrhagic shock homogenized regional myocardial perfusion in coronary stenosis and that fluid therapy failed to restore this. PMID- 9390953 TI - Redox behavior of cytochrome oxidase in the rat brain measured by near-infrared spectroscopy. AB - Using near-infrared spectroscopy, we developed a new approach for measuring the redox state of cytochrome oxidase in the brain under normal blood-circulation conditions. Our algorithm does not require the absorption coefficient of cytochrome oxidase, which differs from study to study. We employed this method for evaluation of effects of changes in oxygen delivery on cerebral oxygenation in rats. When fractional inspired oxygen was decreased in a stepwise manner from 100 to <10%, at which point the concentration of oxygenated hemoglobin ([HbO2]) decreased by approximately 60%, cytochrome oxidase started to be reduced. Increases in arterial PO2 under hyperoxic conditions caused an increase in [HbO2], whereas further oxidation of cytochrome oxidase was not observed. The dissociation of the responses of hemogloblin and cytochrome oxidase was also clearly observed after the injection of epinephrine under severely hypoxic conditions; that is, cytochrome oxidase was reoxidized with increasing blood pressure, whereas hemoglobin oxygenation was not changed. These data indicated that oxygen-dependent redox changes in cytochrome oxidase occur only when oxygen delivery is extremely impaired. This is consistent with the in vitro data of our previous study. PMID- 9390954 TI - Effects of surfactant proteins SP-B and SP-C on dynamic and static mechanics of immature lungs. AB - To investigate the effects of surfactant proteins B (SP-B) and C (SP-C) on lung mechanics, we compared tidal and static lung volumes of immature rabbits anesthetized with pentobarbital sodium and given reconstituted test surfactants (RTS). With a series of RTS having various SP-B concentrations (0-0.7%) but a fixed SP-C concentration (1.4%), both the tidal volume with 25-cmH2O insufflation pressure and the static volume deflated to 5-cmH2O airway pressure increased, significantly correlating with the SP-B concentration: the former increased from 6.5 to 26.0 ml/kg (mean), and the latter increased from 6.4 to 31.8 ml/kg. With another series of RTS having a fixed SP-B concentration (0.7%) but various SP-C concentrations (0-1.4%), the tidal volume increased from 5.1 to 24.8 ml/kg, significantly correlating with the SP-C concentration, whereas the static volume increased from 3.4 to 32.0 ml/kg, the ceiling value, in the presence of a minimal concentration of SP-C (0. 18%). In conclusion, certain doses of SP-B and SP-C were indispensable for optimizing dynamic lung mechanics; the static mechanics, however, required significantly less SP-C. PMID- 9390955 TI - Growth hormone/IGF-I and/or resistive exercise maintains myonuclear number in hindlimb unweighted muscles. AB - In the present study of rats, we examined the role, during 2 wk of hindlimb suspension, of growth hormone/insulin-like growth factor I (GH/IGF-I) administration and/or brief bouts of resistance exercise in ameliorating the loss of myonuclei in fibers of the soleus muscle that express type I myosin heavy chain. Hindlimb suspension resulted in a significant decrease in mean soleus wet weight that was attenuated either by exercise alone or by exercise plus GH/IGF-I treatment but was not attenuated by hormonal treatment alone. Both mean myonuclear number and mean fiber cross-sectional area (CSA) of fibers expressing type I myosin heavy chain decreased after 2 wk of suspension compared with control (134 vs. 162 myonuclei/mm and 917 vs. 2,076 micron2, respectively). Neither GH/IGF-I treatment nor exercise alone affected myonuclear number or fiber CSA, but the combination of exercise and growth-factor treatment attenuated the decrease in both variables. A significant correlation was found between mean myonuclear number and mean CSA across all groups. Thus GH/IGF-I administration and brief bouts of muscle loading had an interactive effect in attenuating the loss of myonuclei induced by chronic unloading. PMID- 9390956 TI - Atrial distension in humans during microgravity induced by parabolic flights. AB - The hypothesis was tested that human cardiac filling pressures increase and the left atrium is distended during 20-s periods of microgravity (microG) created by parabolic flights, compared with values of the 1-G supine position. Left atrial diameter (n = 8, echocardiography) increased significantly during microG from 26.8 +/- 1.2 to 30.4 +/- 0.7 mm (P < 0.05). Simultaneously, central venous pressure (CVP; n = 6, transducer-tipped catheter) decreased from 5.8 +/- 1.5 to 4.5 +/- 1.1 mmHg (P < 0.05), and esophageal pressure (EP; n = 6) decreased from 1.5 +/- 1.6 to -4.1 +/- 1.7 mmHg (P < 0.05). Thus transmural CVP (TCVP = CVP - EP; n = 4) increased during microG from 6.1 +/- 3. 2 to 10.4 +/- 2.7 mmHg (P < 0.05). It is concluded that short periods of microG during parabolic flights induce an increase in TCVP and left atrial diameter in humans, compared with the results obtained in the 1-G horizontal supine position, despite a decrease in CVP. PMID- 9390957 TI - Regional intramuscular pressure development and fatigue in the canine gastrocnemius muscle in situ. AB - Intramuscular pressure (PIM) was measured simultaneously in zones of the medial head of the gastrocnemius-plantaris muscle group (zone I, popliteal origin; zone II, central; zone III, near calcaneus tendon) to determine regional muscle mechanics during isometric tetanic contractions. Peak PIM averages were 586, 1,676, and 993 mmHg deep in zones I, II, and III and 170, 371, and 351 mmHg superficially in zones I, II, and III, respectively. During fatigue, loss of PIM across zones was greatest in zone III (-81%) and least in zone I (-60%) when whole muscle tension loss was -49%. Recovery of PIM was greatest in zone III and least in zone II, achieving 86% and 67% of initial PIM, respectively, when tension recovered to 89%. These data demonstrate that 1) regional mechanical performance can be measured as PIM within a whole muscle, 2) PIM is nonuniform within the canine gastrocnemius-plantaris muscle, being greatest in the deep central zone, and 3) fatigue and recovery of PIM are dissimilar across regions. These differences suggest distinct local effects that integrate to determine whole muscle mechanical capacity during and after intense exercise. PMID- 9390958 TI - Postexercise protein-carbohydrate and carbohydrate supplements increase muscle glycogen in men and women. AB - We have previously demonstrated that women did not increase intramuscular glycogen in response to an increased percent of dietary carbohydrate (CHO) (from 60 to 75% of energy intake) (M. A. Tarnopolsky, S. A. Atkinson, S. M. Phillips, and J. D. MacDougall. J. Appl. Physiol. 78: 1360-1368, 1995). CHO and CHO-protein (Pro) supplementation postexercise can potentiate glycogen resynthesis compared with placebo (K. M. Zawadzki, B. B. Yaspelkis, and J. L. Ivy. J. Appl. Physiol. 72: 1854-1859, 1992). We studied the effect of isoenergetic CHO and CHO-Pro-Fat supplements on muscle glycogen resynthesis in the first 4 h after endurance exercise (90 min at 65% peak O2 consumption) in trained endurance athletes (men, n = 8; women, tested in midfollicular phase, n = 8). Each subject completed three sequential trials separated by 3 wk; a supplement was provided immediately and 1 h postexercise: 1) CHO (0.75 g/kg) + Pro (0.1 g/kg) + Fat (0.02 g/kg), 2) CHO (1 g/kg), and 3) placebo (Pl; artificial sweetener). Subjects were given prepackaged, isoenergetic, isonitrogenous diets, individualized to their habitual diet, for the day before and during the exercise trial. During exercise, women oxidized more lipid than did men (P < 0.05). Both of the supplement trials resulted in greater postexercise glucose and insulin compared with Pl (P < 0.01), with no gender differences. Similarly, both of these trials resulted in increased glycogen resynthesis (37.2 vs. 24. 6 mmol . kg dry muscle-1 . h-1, CHO vs. CHO Pro-Fat, respectively) compared with Pl (7.5 mmol . kg dry muscle-1 . h-1; P < 0.001) with no gender differences. We conclude that postexercise CHO and CHO-Pro Fat nutritional supplements can increase glycogen resynthesis to a greater extent than Pl for both men and women. PMID- 9390959 TI - Hyperventilation-induced airway injury and vascular leakage in dogs: effects of alpha1-adrenergic agonists. AB - alpha1-Adrenergic agonists inhibit hyperventilation-induced bronchoconstriction (HIB) in dogs. We tested the hypothesis that alpha-agonists inhibit HIB by reducing bronchovascular leakage and edema that theoretically could cause airway obstruction. Peripheral airways were isolated by using a bronchoscope; pretreated with either methoxamine (Mx), norepinephrine (NE), or saline aerosol; and then exposed to a 2,000 ml/min dry-air challenge (DAC) for 2 min. Colloidal carbon was injected before DAC and used to quantify bronchovascular permeability. Mx-, NE-, and vehicle-treated airways were prepared for morphometric analysis within 1 h after DAC. Light microscopy revealed that the 2-min DAC produced minimal bronchovascular leakage and little epithelial damage. However, pretreatment with either Mx or NE significantly enhanced dry air-induced bronchovascular hyperpermeability and mucosal injury. The increased damage associated with these alpha1-agonists implicates a protective role for the bronchial circulation. The fact that alpha1-agonists inhibit HIB suggests that neither dry air-induced leakage nor injury directly contributes to the development of airway obstruction. In addition, our data suggest that alpha-agonists attenuate HIB in part by augmenting hyperventilation-induced bronchovascular leakage and by replacing airway water lost during a DAC. PMID- 9390960 TI - Intensity and frequency dependence of laryngeal afferent inputs to respiratory hypoglossal motoneurons. AB - Inspiratory hypoglossal motoneurons (IHMs) mediate contraction of the genioglossus muscle and contribute to the regulation of upper airway patency. Intracellular recordings were obtained from antidromically identified IHMs in anesthetized, vagotomized cats, and IHM responses to electrical activation of superior laryngeal nerve (SLN) afferent fibers at various frequencies and intensities were examined. SLN stimulus frequencies <2 Hz evoked an excitatory inhibitory postsynaptic potential (EPSP-IPSP) sequence or only an IPSP in most IHMs that did not change in amplitude as the stimulus was maintained. During sustained stimulus frequencies of 5-10 Hz, there was a reduction in the amplitude of SLN-evoked IPSPs with time with variable changes in the EPSP. At stimulus frequencies >25 Hz, the amplitude of EPSPs and IPSPs was reduced over time. At a given stimulus frequency, increasing stimulus intensity enhanced the decay of the SLN-evoked postsynaptic potentials (PSPs). Frequency-dependent attenuation of SLN inputs to IHMs also occurred in newborn kittens. These results suggest that activation of SLN afferents evokes different PSP responses in IHMs depending on the stimulus frequency. At intermediate frequencies, inhibitory inputs are selectively filtered so that excitatory inputs predominate. At higher frequencies there was no discernible SLN-evoked PSP temporally locked to the SLN stimuli. Alterations in SLN-evoked PSPs could play a role in the coordination of genioglossal contraction during respiration, swallowing, and other complex motor acts where laryngeal afferents are activated. PMID- 9390961 TI - Cardiovascular adaptations to 10 days of cycle exercise. AB - We hypothesized that 10 days of training would enhance cardiac output (CO) and stroke volume (SV) during peak exercise and increase the inotropic response to beta-adrenergic stimulation. Ten subjects [age 26 +/- 2 (SE) yr] trained on a cycle ergometer for 10 days. At peak exercise, training increased O2 uptake, CO, and SV (P < 0.001). Left ventricular (LV) size and function at rest were assessed with two-dimensional echocardiography before (baseline) and after atropine injection (1.0 mg) and during four graded doses of dobutamine. LV end-diastolic diameter increased with training (P < 0.02), whereas LV wall thickness was unchanged. LV contractile performance was assessed by relating fractional shortening (FS) to the estimated end-systolic wall stress (sigmaES). Training increased the slope of the FS-sigmaES relationship (P < 0.05), indicating enhanced systolic function. The increase in slope correlated with increases in CO (r = -0.71, P < 0.05) and SV (r = -0.70, P < 0.05). The increase in blood volume also correlated with increases in CO (r = 0.80, P < 0.01) and SV (r = 0.85, P < 0.004). These data show that 10 days of training enhance the inotropic response to beta-adrenergic stimulation, associated with increases in CO and SV during peak exercise. PMID- 9390962 TI - Ventilation distribution during histamine provocation. AB - We investigated ventilation inhomogeneity during provocation with inhaled histamine in 20 asymptomatic nonsmoking subjects. We used N2 multiple-breath washout (MBW) to derive parameters Scond and Sacin as a measurement of ventilation inhomogeneity in conductive and acinar zones of the lungs, respectively. A 20% decrease of forced expiratory volume in 1 s (FEV1) was used to distinguish responders from nonresponders. In the responder group, average FEV1 decreased by 26%, whereas Scond increased by 390% with no significant change in Sacin. In the nonresponder group, FEV1 decreased by 11%, whereas Scond increased by 198% with no significant Sacin change. Despite the absence of change in Sacin during provocation, baseline Sacin was significantly larger in the responder vs. the nonresponder group. The main findings of our study are that during provocation large ventilation inhomogeneities occur, that the small airways affected by the provocation process are situated proximal to the acinar zone where the diffusion front stands, and that, in addition to overall decrease in airway caliber, there is inhomogeneous narrowing of parallel airways. PMID- 9390963 TI - Endurance training attenuates the decrease in skeletal muscle malonyl-CoA with exercise. AB - Muscle malonyl-CoA has been postulated to regulate fatty acid metabolism by inhibiting carnitine palmitoyltransferase 1. In nontrained rats, malonyl-CoA decreases in working muscle during exercise. Endurance training is known to increase a muscle's reliance on fatty acids as a substrate. This study was designed to investigate whether the decline in malonyl-CoA with exercise would be greater in trained than in nontrained muscle, thereby allowing increased fatty acid oxidation. After 6-10 wk of endurance training (2 h/day) or treadmill habituation (5-10 min/day), rats were killed at rest or after running up a 15% grade at 21 m/min for 5, 20, or 60 min. Training attenuated the exercise-induced drop in malonyl-CoA and prevented the exercise-induced increase in the constant for citrate activation of acetyl-CoA carboxylase in the red quadriceps muscle of rats run for 20 and 60 min. Hence, contrary to expectations, the decrease in malonyl-CoA was less in trained than in nontrained muscle during a single bout of prolonged submaximal exercise. PMID- 9390964 TI - Neural-mechanical coupling of breathing in REM sleep. AB - During rapid-eye-movement (REM) sleep the ventilatory response to airway occlusion is reduced. Possible mechanisms are reduced chemosensitivity, mechanical impairment of the chest wall secondary to the atonia of REM sleep, or phasic REM events that interrupt or fractionate ongoing diaphragm electromyogram (EMG) activity. To differentiate between these possibilities, we studied three chronically instrumented dogs before, during, and after 15-20 s of airway occlusion during non-REM (NREM) and phasic REM sleep. We found that 1) for a given inspiratory time the integrated diaphragm EMG (Di) was similar or reduced in REM sleep relative to NREM sleep; 2) for a given Di in response to airway occlusion and the hyperpnea following occlusion, the mechanical output (flow or pressure) was similar or reduced during REM sleep relative to NREM sleep; 3) for comparable durations of airway occlusion the Di and integrated inspiratory tracheal pressure tended to be smaller and more variable in REM than in NREM sleep, and 4) significant fractionations (caused visible changes in tracheal pressure) of the diaphragm EMG during airway occlusion in REM sleep occurred in approximately 40% of breathing efforts. Thus reduced and/or erratic mechanical output during and after airway occlusion in REM sleep in terms of flow rate, tidal volume, and/or pressure generation is attributable largely to reduced neural activity of the diaphragm, which in turn is likely attributable to REM effects, causing reduced chemosensitivity at the level of the peripheral chemoreceptors or, more likely, at the central integrator. Chest wall distortion secondary to the atonia of REM sleep may contribute to the reduced mechanical output following airway occlusion when ventilatory drive is highest. PMID- 9390965 TI - Prostaglandin production contributes to exercise-induced vasodilation in heart failure. AB - Endothelial release of prostaglandins may contribute to exercise-induced skeletal muscle arteriolar vasodilation in patients with heart failure. To test this hypothesis, we examined the effect of indomethacin on leg circulation and metabolism in eight chronic heart failure patients, aged 55 +/- 4 yr. Central hemodynamics and leg blood flow, determined by thermodilution, and leg metabolic parameters were measured during maximum treadmill exercise before and 2 h after oral administration of indomethacin (75 mg). Leg release of 6-ketoprostaglandin F1alpha was also measured. During control exercise, leg blood flow increased from 0.34 +/- 0.03 to 1. 99 +/- 0.19 l/min (P < 0.001), leg O2 consumption from 13.6 +/- 1.8 to 164.5 +/- 16.2 ml/min (P < 0.001), and leg prostanoid release from 54.1 +/- 8.5 to 267.4 +/- 35.8 pg/min (P < 0.001). Indomethacin suppressed release of prostaglandin F1alpha (P < 0.001) throughout exercise and decreased leg blood flow during exercise (P < 0.05). This was associated with a corresponding decrease in leg O2 consumption (P < 0.05) and a higher level of femoral venous lactate at peak exercise (P < 0.01). These data suggest that release of vasodilatory prostaglandins contributes to skeletal muscle arteriolar vasodilation in patients with heart failure. PMID- 9390966 TI - Nitric oxide and endothelial permeability. AB - Nitric oxide synthase inhibition reverses systemic vasodilation during sepsis but may increase endothelial permeability. To assess adverse effects on the pulmonary vasculature, 12 sheep were chronically instrumented with lung lymph fistulas and hydraulic pulmonary venous occluders. Escherichia coli endotoxin (lipopolysaccharide; 10 ng . kg-1 . min-1) was continuously infused for 32 h. After 24 h, six animals received 25 mg/kg of Nomega-nitro-L-arginine methyl ester (L-NAME), and six received saline. All sheep developed a hyperdynamic circulatory response and elevated lymph flows by 24 h of lipopolysaccharide infusion. L-NAME reversed systemic vasodilation, increased pre- and postcapillary pulmonary vascular resistance index, pulmonary arterial pressure, and, transiently, effective pulmonary capillary pressure. Lung lymph flows were not different between groups at 24 h or thereafter. Calculated as changes from baseline, however, lung lymph flow was higher in the L-NAME group than in the control animals, with a trend toward lower lymph-to-plasma protein concentration ratio at 25 h. Permeability analysis at 32 h by the venous occlusion technique showed normal reflection coefficients and elevated filtration coefficients without differences between groups. Reversal by L-NAME of the systemic vasodilation during endotoxemia was associated with high pulmonary vascular resistance without evidence of impaired pulmonary endothelial barrier function. PMID- 9390967 TI - Greater rate of decline in maximal aerobic capacity with age in physically active vs. sedentary healthy women. AB - Using a meta-analytic approach, we recently reported that the rate of decline in maximal oxygen uptake (VO2 max) with age in healthy women is greatest in the most physically active and smallest in the least active when expressed in milliliters per kilogram per minute per decade. We tested this hypothesis prospectively under well-controlled laboratory conditions by studying 156 healthy, nonobese women (age 20-75 yr): 84 endurance-trained runners (ET) and 72 sedentary subjects (S). ET were matched across the age range for age-adjusted 10-km running performance. Body mass was positively related with age in S but not in ET. Fat-free mass was not different with age in ET or S. Maximal respiratory exchange ratio and rating of perceived exertion were similar across age in ET and S, suggesting equivalent voluntary maximal efforts. There was a significant but modest decline in running mileage, frequency, and speed with advancing age in ET. VO2 max (ml . kg-1 . min 1) was inversely related to age (P < 0.001) in ET (r = -0.82) and S (r = -0.71) and was higher at any age in ET. Consistent with our meta-analysic findings, the absolute rate of decline in VO2 max was greater in ET (-5.7 ml . kg-1 . min-1 . decade-1) compared with S (-3.2 ml . kg-1 . min-1 . decade-1; P < 0. 01), but the relative (%) rate of decline was similar (-9.7 vs -9. 1%/decade; not significant). The greater absolute rate of decline in VO2 max in ET compared with S was not associated with a greater rate of decline in maximal heart rate (-5.6 vs. -6.2 beats . min-1 . decade-1), nor was it related to training factors. The present cross-sectional findings provide additional evidence that the absolute, but not the relative, rate of decline in maximal aerobic capacity with age may be greater in highly physically active women compared with their sedentary healthy peers. This difference does not appear to be related to age-associated changes in maximal heart rate, body composition, or training factors. PMID- 9390968 TI - Cardiopulmonary control in sleeping Sprague-Dawley rats treated with hydralazine. AB - To test the hypothesis that hydralazine can suppress spontaneous sleep-related central apnea, respiratory pattern, blood pressure, and heart period were monitored in Sprague-Dawley rats. In random order and on separate days, rats were recorded after intraperitoneal injection of 1) saline or 2) 2 mg/kg hydralazine. Normalized minute ventilation (NVI) declined significantly with transitions from wake to non-rapid-eye-movement (NREM) sleep (-5.1%; P = 0.01) and rapid-eye movement (REM) sleep (-4.2%; P = 0.022). Hydralazine stimulated respiration (NVI increased by 21%; P < 0.03) and eliminated the effect of state on NVI. Blood pressure decreased by 17% after hydralazine, and the correlation between fluctuations in mean blood pressure and NVI changed from strongly positive during control recordings to weakly negative after hydralazine (P < 0.0001 for each). Postsigh and spontaneous apneas were reduced during NREM and REM sleep after hydralazine (P < 0.05 for each). This suppression was strongly correlated with the reduction in blood pressure and with the degree of respiratory stimulation. We conclude that mild hydralazine-induced hypotension leads to respiratory stimulation and apnea suppression. PMID- 9390969 TI - Isoproterenol attenuates high vascular pressure-induced permeability increases in isolated rat lungs. AB - To separate the contributions of cellular and basement membrane components of the alveolar capillary barrier to the increased microvascular permeability induced by high pulmonary venous pressures (Ppv), we subjected isolated rat lungs to increases in Ppv, which increased capillary filtration coefficient (Kfc) without significant hemorrhage (31 cmH2O) and with obvious extravasation of red blood cells (43 cmH2O). Isoproterenol (20 microM) was infused in one group (Iso) to identify a reversible cellular component of injury, and residual blood volumes were measured to assess extravasation of red blood cells through ruptured basement membranes. In untreated lungs (High Ppv group), Kfc increased 6.2 +/- 1.3 and 38.3 +/- 15.2 times baseline during the 31 and 43 cmH2O Ppv states. In Iso lungs, Kfc was 36.2% (P < 0.05) and 64.3% of that in the High Ppv group at these Ppv states. Residual blood volumes calculated from tissue hemoglobin contents were significantly increased by 53-66% in the high Ppv groups, compared with low vascular pressure controls, but there was no significant difference between High Ppv and Iso groups. Thus isoproterenol significantly attenuated vascular pressure-induced Kfc increases at moderate Ppv, possibly because of an endothelial effect, but it did not affect red cell extravasation at higher vascular pressures. PMID- 9390970 TI - Intratracheal instillation of a novel NO/nucleophile adduct selectively reduces pulmonary hypertension. AB - We examined the pulmonary and systemic hemodynamic effects of administering soluble nitric oxide (NO) donor compounds (NO/nucleophile adducts, i.e., NONOates) directly into the trachea of animals with experimentally induced pulmonary hypertension. Steady-state pulmonary hypertension was created by using the thromboxane agonist U-46619. Yorkshire pigs were randomly assigned to one of four groups: group 1, intratracheal saline (control; n = 8); group 2, intratracheal sodium nitroprusside (n = 6); group 3, intratracheal ethylputreanine NONOate (n = 6); and group 4, intratracheal 2-(dimethylamino) ethylputreanine NONOate (DMAEP/NO; n = 6). Pulmonary and systemic hemodynamics were monitored after drug instillation. Group 4 had significant reductions in pulmonary vascular resistance index (PVRI) at all time points compared with steady state and compared with group 1 (P < 0.05), whereas systemic vascular resistance index did not change. The mean change in mean pulmonary arterial pressure in group 4 was -33.1 +/- 1.2% compared with +6.4 +/- 1.3% in group 1 (P < 0.001), and the mean change in mean arterial pressure was -9.3 +/- 0.7% compared with a control value of -0.9 +/- 0.5% (P < 0.05). Groups 2 and 3 had significant decreases in both PVRI and systemic vascular resistance index compared with steady state and with group 1. In conclusion, intratracheal instillation of a polar-charged tertiary amine NONOate DMAEP/NO results in the selective reduction of PVRI. Intermittent intratracheal instillation of selective NONOates may be an alternative to continuously inhaled NO in the treatment of pulmonary hypertension. PMID- 9390971 TI - Optical measurement of isolated canine lung filtration coefficients at normal hematocrits. AB - In this study, lung filtration coefficient (Kfc) values were measured in eight isolated canine lung preparations at normal hematocrit values using three methods: gravimetric, blood-corrected gravimetric, and optical. The lungs were kept in zone 3 conditions and subjected to an average venous pressure increase of 10.24 +/- 0.27 (SE) cmH2O. The resulting Kfc (ml . min-1 . cmH2O-1 . 100 g dry lung wt-1) measured with the gravimetric technique was 0.420 +/- 0.017, which was statistically different from the Kfc measured by the blood-corrected gravimetric method (0.273 +/- 0.018) or the product of the reflection coefficient (sigmaf) and Kfc measured optically (0. 272 +/- 0.018). The optical method involved the use of a Cellco filter cartridge to separate red blood cells from plasma, which allowed measurement of the concentration of the tracer in plasma at normal hematocrits (34 +/- 1.5). The permeability-surface area product was measured using radioactive multiple indicator-dilution methods before, during, and after venous pressure elevations. Results showed that the surface area of the lung did not change significantly during the measurement of Kfc. These studies suggest that sigmafKfc can be measured optically at normal hematocrits, that this measurement is not influenced by blood volume changes that occur during the measurement, and that the optical sigmafKfc agrees with the Kfc obtained via the blood-corrected gravimetric method. PMID- 9390972 TI - Gender differences in airway resistance during sleep. AB - At the onset of non-rapid-eye-movement (NREM) sleep there is a fall in ventilation and an increase in upper airway resistance (UAR). In healthy men there is a progressive increase in UAR as NREM sleep deepens. This study compared the pattern of change in UAR and ventilation in 14 men and 14 women (aged 18-25 yr) both during sleep onset and over the NREM phase of a sleep cycle (from wakefulness to slow-wave sleep). During sleep onset, fluctuations between electroencephalographic alpha and theta activity were associated with mean alterations in inspiratory minute ventilation and UAR of between 1 and 4.5 l/min and between 0.70 and 5.0 cmH2O . l-1 . s, respectively, with no significant effect of gender on either change (P > 0.05). During NREM sleep, however, the increment in UAR was larger in men than in women (P < 0.01), such that the mean levels of UAR at peak flow reached during slow-wave sleep were approximately 25 and 10 cmH2O . l-1 . s in men and women, respectively. We speculate that the greater increase in UAR in healthy young men may represent a gender-related susceptibility to sleep-disordered breathing that, in conjunction with other predisposing factors, may contribute to the development of obstructive sleep apnea. PMID- 9390973 TI - Strength, skeletal muscle composition, and enzyme activity in multiple sclerosis. AB - This study examined functional, biochemical, and morphological characteristics of skeletal muscle in nine multiple sclerosis (MS) patients and eight healthy controls in an effort to ascertain whether intramuscular adaptations could account for excessive fatigue in this disease. Analyses of biopsies of the tibialis anterior muscle showed that there were fewer type I fibers (66 +/- 6 vs. 76 +/- 6%), and that fibers of all types were smaller (average downward arrow26%) and had lower succinic dehydrogenase (SDH; average downward arrow40%) and SDH/alpha-glycerol-phosphate dehydrogenase (GPDH) but not GPDH activities in MS vs. control subjects, suggesting that muscle in this disease is smaller and relies more on anaerobic than aerobic-oxidative energy supply than does muscle of healthy individuals. Maximal voluntary isometric force for dorsiflexion was associated with both average fiber cross-sectional area (r = 0.71, P = 0.005) and muscle fat-free cross-sectional area by magnetic resonance imaging (r = 0.80, P < 0. 001). Physical activity, assessed by accelerometer, was associated with average fiber SDH/GPDH (r = 0.78, P = 0.008). There was a tendency for symptomatic fatigue to be inversely associated with average fiber SDH activity (r = -0.57, P = 0.068). The results of this study suggest that the inherent characteristics of skeletal muscle fibers per se and of skeletal muscle as a whole are altered in the direction of disuse in MS. They also suggest that changes in skeletal muscle in MS may significantly affect function. PMID- 9390974 TI - Correlation between ventilation and EEG-defined arousal during sleep onset in young subjects. AB - In studies of elderly individuals, ventilation and EEG-defined arousal have been shown to vary periodically and synchronously. Such results have been interpreted as indicating the primacy of sleep/wake state in causing ventilatory instability during sleep onset. However, because the elderly individuals studied were periodic breathers, the results do not unequivocally support this conclusion. In this study the relationship between ventilation and EEG-defined arousal was assessed in a group of 21 young, healthy men in whom ventilatory instability during sleep onset was not periodic. Ventilation and EEG (O1-A2) recordings were collected, and the longest uncontaminated periods from early and late in sleep onset were selected for subsequent analysis. The 84 time series (21 subjects, 2 variables, and 2 occasions in sleep onset) were subjected to spectral analysis to identify periodicity, and the relationship between the two variables was determined by cross-correlational methods. The results indicated that the time series were nonperiodic, yet significant correlations were observed between the two variables. The data support the view that during sleep onset ventilatory instability is driven primarily by variations in sleep/wake arousal level. PMID- 9390975 TI - Modification of active cutaneous vasodilation by oral contraceptive hormones. AB - It is not clear whether the altered thermoregulatory reflex control of the cutaneous circulation seen among phases of the menstrual cycle also occurs with the synthetic estrogen and progesterone in oral contraceptive pills and whether any such modifications include altered control of the cutaneous active vasodilator system. To address these questions, we conducted controlled whole body heating experiments in seven women at the end of the third week of hormone pills (HH) and at the end of the week of placebo/no pills (LH). A water-perfused suit was used to control body temperature. Laser Doppler flowmetry was used to monitor cutaneous blood flow at a control site and at a site at which noradrenergic vasoconstrictor control had been eliminated by iontophoresis of bretylium (BT), isolating the active cutaneous vasodilator system. The oral temperature (Tor) thresholds for cutaneous vasodilation were higher in HH at both control [37.09 +/- 0.12 vs. 36.83 +/- 0.07 degrees C (LH), P < 0.01] and BT treated [37. 19 +/- 0.05 vs. 36.88 +/- 0.12 degrees C (LH), P < 0.01] sites. The Tor threshold for sweating was similarly shifted (HH: 37.15 +/- 0.11 degrees C vs. LH: 36.94 +/- 0.11 degrees C, P < 0.01). A rightward shift in the relationship of heart rate to Tor was seen in HH. The sensitivities (slopes of the responses vs. Tor) did not differ statistically between phases. The similar threshold shifts at control and BT-treated sites suggest that the hormones shift the function of the active vasodilator system to higher internal temperatures. The similarity of the shifts among thermoregulatory effectors suggests a centrally mediated action of these hormones. PMID- 9390976 TI - Effects of training and a single session of exercise on lipids and apolipoproteins in hypercholesterolemic men. AB - To differentiate between transient (acute) and training (chronic) effects of exercise at two different intensities on blood lipids and apolipoproteins (apo), 26 hypercholesterolemic men (cholesterol = 258 mg/dl, age = 47 yr, weight = 81.9 kg) trained three times per week for 24 wk, 350 kcal/session at high (80% maximal O2 uptake, n = 12) or moderate (50% maximal O2 uptake, n = 14) intensity. Serum lipid and apolipoprotein (apo) concentrations (plasma volume adjusted) were measured before and immediately, 24, and 48 h after exercise on four different occasions corresponding to 0, 8, 16, and 24 wk of training. Data were analyzed using three-way repeated-measures multivariate analysis of variance followed by analysis of variance and Duncan's procedures (alpha = 0.05). A transient 6% rise in low-density-lipoprotein cholesterol measured before training at the 24-h time point was no longer evident after training. Triglycerides fell and total cholesterol, high-density-lipoprotein cholesterol (HDL-C), HDL3-C, apo A-I, and apo B rose 24-48 h after exercise regardless of training or intensity. Total cholesterol, HDL3-C, apo A-I, and apo B were lower and HDL2-C was higher after training than before training. Thus exercise training and a single session of exercise exert distinct and interactive effects on lipids and apolipoproteins. These results support the practice of training at least every other day to obtain optimal exercise benefits. PMID- 9390977 TI - Effect of microgravity and hypergravity on deposition of 0.5- to 3-micron diameter aerosol in the human lung. AB - We measured intrapulmonary deposition of 0. 5-, 1-, 2-, and 3-micron-diameter particles in four subjects on the ground (1 G) and during parabolic flights both in microgravity (microG) and at approximately 1.6 G. Subjects breathed aerosols at a constant flow rate (0.4 l/s) and tidal volume (0.75 liter). At 1 G and approximately 1.6 G, deposition increased with increasing particle size. In microG, differences in deposition as a function of particle size were almost abolished. Deposition was a nearly linear function of the G level for 2- and 3 micron-diameter particles, whereas for 0.5- and 1.0-micron-diameter particles, deposition increased less between microG and 1 G than between 1 G and approximately 1.6 G. Comparison with numerical predictions showed good agreement for 1-, 2-, and 3-micron-diameter particles at 1 and approximately 1.6 G, whereas the model consistently underestimated deposition in microG. The higher deposition observed in microG compared with model predictions might be explained by a larger deposition by diffusion because of a higher alveolar concentration of aerosol in microG and to the nonreversibility of the flow, causing additional mixing of the aerosols. PMID- 9390978 TI - Autonomic control of skeletal muscle vasodilation during exercise. AB - Despite extensive investigation, the control of blood flow during dynamic exercise is not fully understood. The purpose of this study was to determine whether beta-adrenergic or muscarinic receptors are involved in the vasodilation in exercising skeletal muscle. Six mongrel dogs were instrumented with ultrasonic flow probes on both external iliac arteries and with a catheter in a branch of one femoral artery. The dogs exercised on a treadmill at 6 miles/h while drugs were injected intra-arterially into one hindlimb. Isoproterenol (0.2 microg) or acetylcholine (1 microg) elicited increases in iliac blood flow of 89.8 +/- 14.4 and 95.6 +/- 17.4%, respectively, without affecting systemic blood pressure or blood flow in the contralateral iliac artery. Intra-arterial propranolol (1 mg) or atropine (500 microg) had no effect on iliac blood flow, although they abolished the isoproterenol and acetylcholine-induced increases in iliac blood flow. These data indicate that exogenous activation of beta-adrenergic or muscarinic receptors in the hindlimb vasculature increases blood flow to dynamically exercising muscle. More importantly, because neither propranolol nor atropine affected iliac blood flow, we conclude that beta-adrenergic and muscarinic receptors are not involved in the control of blood flow to skeletal muscle during moderate steady-state dynamic exercise in dogs. PMID- 9390979 TI - Changes in insulin-stimulated glucose transport and GLUT-4 protein in rat skeletal muscle after training. AB - After running training, which increased GLUT-4 protein content in rat skeletal muscle by <40% compared with control rats, the training effect on insulin stimulated maximal glucose transport (insulin responsiveness) in skeletal muscle was short lived (24 h). A recent study reported that GLUT-4 protein content in rat epitrochlearis muscle increased dramatically ( approximately 2-fold) after swimming training (J.-M. Ren, C. F. Semenkovich, E. A. Gulve, J. Gao, and J. O. Holloszy. J. Biol. Chem. 269, 14396-14401, 1994). Because GLUT-4 protein content is known to be closely related to skeletal muscle insulin responsiveness, we thought it possible that the training effect on insulin responsiveness may remain for >24 h after swimming training if GLUT-4 protein content decreases gradually from the relatively high level and still remains higher than control level for >24 h after swimming training. Therefore, we examined this possibility. Male Sprague-Dawley rats swam 2 h a day for 5 days with a weight equal to 2% of body mass. Approximately 18, 42, and 90 h after cessation of training, GLUT-4 protein concentration and 2-[1,2-3H]deoxy-D-glucose transport in the presence of a maximally stimulating concentration of insulin (2 mU/ml) were examined by using incubated epitrochlearis muscle preparation. Swimming training increased GLUT-4 protein concentration and insulin responsiveness by 87 and 85%, respectively, relative to age-matched controls when examined 18 h after training. Forty-two hours after training, GLUT-4 protein concentration and insulin responsiveness were still higher by 52 and 51%, respectively, in muscle from trained rats compared with control. GLUT-4 protein concentration and insulin responsiveness in trained muscle returned to sedentary control level within 90 h after training. We conclude that 1) the change in insulin responsiveness during detraining is directly related to muscle GLUT-4 protein content, and 2) consequently, the greater the increase in GLUT-4 protein content that is induced by training, the longer an effect on insulin responsiveness persists after the training. PMID- 9390980 TI - Neural mechanism of the pressor response to obstructive and nonobstructive apnea. AB - Obstructive and nonobstructive apneas elicit substantial increases in muscle sympathetic nerve activity and arterial pressure. The time course of change in these variables suggests a causal relationship; however, mechanical influences, such as release of negative intrathoracic pressure and reinflation of the lungs, are potential contributors to the arterial pressure rise. To test the hypothesis that apnea-induced pressor responses are neurally mediated, we measured arterial pressure (photoelectric plethysmography), muscle sympathetic nerve activity (peroneal microneurography), arterial O2 saturation (pulse oximeter), and end tidal CO2 tension (gas analyzer) during sustained Mueller maneuvers, intermittent Mueller maneuvers, and simple breath holds in six healthy humans before, during, and after ganglionic blockade with trimethaphan (3-4 mg/min, titrated to produce complete disappearance of sympathetic bursts from the neurogram). Ganglionic blockade abolished the pressor responses to sustained and intermittent Mueller maneuvers (-4 +/- 1 vs. +15 +/- 3 and 0 +/- 2 vs. +15 +/- 5 mmHg) and breath holds (0 +/- 3 vs. +11 +/- 3, all P < 0.05). We conclude that the acute pressor response to obstructive and nonobstructive voluntary apnea is sympathetically mediated. PMID- 9390981 TI - Long-term creatine intake is beneficial to muscle performance during resistance training. AB - The effects of oral creatine supplementation on muscle phosphocreatine (PCr) concentration, muscle strength, and body composition were investigated in young female volunteers (n = 19) during 10 wk of resistance training (3 h/wk). Compared with placebo, 4 days of high-dose creatine intake (20 g/day) increased (P < 0.05) muscle PCr concentration by 6%. Thereafter, this increase was maintained during 10 wk of training associated with low-dose creatine intake (5 g/day). Compared with placebo, maximal strength of the muscle groups trained, maximal intermittent exercise capacity of the arm flexors, and fat-free mass were increased 20-25, 10 25, and 60% more (P < 0. 05), respectively, during creatine supplementation. Muscle PCr and strength, intermittent exercise capacity, and fat-free mass subsequently remained at a higher level in the creatine group than in the placebo group during 10 wk of detraining while low-dose creatine was continued. Finally, on cessation of creatine intake, muscle PCr in the creatine group returned to normal within 4 wk. It is concluded that long-term creatine supplementation enhances the progress of muscle strength during resistance training in sedentary females. PMID- 9390982 TI - A dual-respiration chamber system with automated calibration. AB - This study characterizes respiration chambers with fully automated calibration. The system consists of two 14-m3 pull-type chambers. Care was taken to provide a friendly environment for the subjects, with the possibility of social contact during the experiment. Gas analysis was automated to correct for analyzer drift and barometric pressure variations and to provide ease of use. Methods used for checking the system's performance are described. The gas-analysis repeatability was within 0.002%. Results of alcohol combustion (50-350 ml/min CO2) show an accuracy of 0.5 +/- 2.0 (SD) % for O2 consumption and -0.3 +/- 1.6% for CO2 production for 2- to 24-h experiments. It is concluded that response time is not the main factor with respect to the smallest practical measurement interval (duration); volume, mixing, gas-analysis accuracy, and levels of O2 consumption and CO2 production are at least equally important. The smallest practical interval was 15-25 min, as also found with most chamber systems described in the literature. We chose to standardize 0.5 h as the minimum measurement interval. PMID- 9390983 TI - Lower extremity muscle activation during horizontal and uphill running. AB - To provide more comprehensive information on the extent and pattern of muscle activation during running, we determined lower extremity muscle activation by using exercise-induced contrast shifts in magnetic resonance (MR) images during horizontal and uphill high-intensity (115% of peak oxygen uptake) running to exhaustion (2.0-3.9 min) in 12 young women. The mean percentage of muscle volume activated in the right lower extremity was significantly (P <0.05) greater during uphill (73 +/- 7%) than during horizontal (67 +/- 8%) running. The percentage of 13 individual muscles or groups activated varied from 41 to 90% during horizontal running and from 44 to 83% during uphill running. During horizontal running, the muscles or groups most activated were the adductors (90 +/- 5%), semitendinosus (86 +/- 13%), gracilis (76 +/- 20%), biceps femoris (76 +/- 12%), and semimembranosus (75 +/- 12%). During uphill running, the muscles most activated were the adductors (83 +/- 8%), biceps femoris (79 +/- 7%), gluteal group (79 +/- 11%), gastrocnemius (76 +/- 15%), and vastus group (75 +/- 13%). Compared with horizontal running, uphill running required considerably greater activation of the vastus group (23%) and soleus (14%) and less activation of the rectus femoris (29%), gracilis (18%), and semitendinosus (17%). We conclude that during high intensity horizontal and uphill running to exhaustion, lasting 2-3 min, muscles of the lower extremity are not maximally activated, suggesting there is a limit to the extent to which additional muscle mass recruitment can be utilized to meet the demand for force and energy. Greater total muscle activation during exhaustive uphill than during horizontal running is achieved through an altered pattern of muscle activation that involves increased use of some muscles and less use of others. PMID- 9390984 TI - Effects of chronic exercise and deconditioning on platelet function in women. AB - To investigate the effects of chronic exercise and deconditioning on platelet function in women, 16 healthy sedentary women were divided into control and exercise groups. The exercise group cycled on an ergometer at 50% maximal oxygen consumption for 30 min/day, 5 days/wk, for two consecutive menstrual cycles and then were deconditioned for three menstrual cycles. During this period, platelet adhesiveness on a fibrinogen-coated surface, ADP-induced platelet aggregation and intracellular calcium concentration elevation, guanosine 3',5'-cyclic monophosphate (cGMP) content in platelets, and plasma nitric oxide metabolite levels were measured before and immediately after a progressive exercise test in the midfollicular phase. Our results indicated that, after exercise training, 1) resting heart rates and blood pressures were reduced, and exercise performance was improved; 2) resting platelet function was decreased, whereas plasma nitrite and nitrate levels and platelet cGMP contents were enhanced; and 3) the potentiation of platelet function by acute strenuous exercise was decreased, whereas the increases in plasma nitrite and nitrate levels and platelet cGMP contents were enhanced by acute exercise. Furthermore, deconditioning reversed these training effects. This implies that training-induced platelet functional changes in women in the midfollicular phase may be mediated by nitric oxide. PMID- 9390985 TI - Bed rest suppresses bioassayable growth hormone release in response to muscle activity. AB - Hormonal responses to muscle activity were studied in eight men before (-13 or 12 and -8 or -7 days), during (2 or 3, 8 or 9, and 13 or 14 days) and after (+2 or +3 and +10 or +11 days) 17 days of bed rest. Muscle activity consisted of a series of unilateral isometric plantar flexions, including 4 maximal voluntary contractions (MVCs), 48 contractions at 30% MVC, and 12 contractions at 80% MVC, all performed at a 4:1-s work-to-rest ratio. Blood was collected before and immediately after muscle activity to measure plasma growth hormone by radioimmunoassay (IGH) and by bioassay (BGH) of tibia epiphyseal cartilage growth in hypophysectomized rats. Plasma IGH was unchanged by muscle activity before, during, or after bed rest. Before bed rest, muscle activity increased (P < 0.05) BGH by 66% at -13 or -12 days (2,146 +/- 192 to 3,565 +/- 197 microg/l) and by 92% at -8 or -7 days (2,162 +/- 159 to 4,161 +/- 204 microg/l). After 2 or 3 days of bed rest, there was no response of BGH to the muscle activity, a pattern that persisted through 8 or 9 days of bed rest. However, after 13 or 14 days of bed rest, plasma concentration of BGH was significantly lower after than before muscle activity (2,594 +/- 211 to 2,085 +/- 109 microg/l). After completion of bed rest, muscle activity increased BGH by 31% at 2 or 3 days (1,807 +/- 117 to 2,379 +/- 473 microg/l; P < 0.05), and by 10 or 11 days the BGH response was similar to that before bed rest (1,881 +/- 75 to 4,160 +/- 315 microg/l; P < 0.05). These data demonstrate that the ambulatory state of an individual can have a major impact on the release of BGH, but not IGH, in response to a single bout of muscle activity. PMID- 9390986 TI - Arterial baroreflex modulation of heat-induced vasodilation in the rabbit ear. AB - The purpose of this study was to determine whether nonthermal baroreflexes arising from cardiopulmonary and/or arterial baroreceptors modulate rabbit ear blood flow (EBF) during hyperthermia. Intact and sinoaortic-denervated (SAD) rabbits were chronically instrumented with a Doppler ultrasonic flow probe for measurement of EBF velocity (kHz). During whole body heating in conscious rabbits, EBF and ear vascular conductance (EVC) increased as core temperature increased until maximal plateau levels of EBF and EVC were reached. The maximal plateau level of EVC attained during heat stress was lower in SAD than in intact rabbits. Subsequent intrapericardial administration of procaine at maximal EBF blocked cardiac afferents but did not alter EVC in either animal group. In a second experiment, ramp decreases in mean arterial pressure were produced by vena caval occlusion at maximal EBF. In intact rabbits, EBF and EVC decreased linearly as mean arterial pressure fell, but EBF and EVC did not decrease during vena caval occlusion in SAD rabbits. Thus neither pharmacological nor mechanical unloading of cardiac baroreceptors results in reflex vasoconstriction in the heat stressed rabbit ear. However, baroreflexes arising from arterial baroreceptors may modulate EBF in heat-stressed rabbits. PMID- 9390987 TI - Ventilation and hypoxic ventilatory responsiveness in Chinese-Tibetan residents at 3,658 m. AB - When breathing ambient air at rest at 3,658 m altitude, Tibetan lifelong residents of 3,658 m ventilate as much as newcomers acclimatized to high altitude; they also ventilate more and have greater hypoxic ventilatory responses (HVRs) than do Han ("Chinese") long-term residents at 3,658 m. This suggests that Tibetan ancestry is advantageous in protecting resting ventilation levels during years of hypoxic exposure and is of interest in light of the permissive role of hypoventilation in the development of chronic mountain sickness, which is nearly absent among Tibetans. The existence of individuals with mixed Tibetan-Chinese ancestry (Han-Tibetans) residing at 3,658 m affords an opportunity to test this hypothesis. Eighteen men born in Lhasa, Tibet, China (3,658 m) to Tibetan mothers and Han fathers were compared with 27 Tibetan men and 30 Han men residing at 3,658 m who were previously studied. We used the same study procedures (minute ventilation was measured with a dry-gas flowmeter during room air breathing and hyperoxia and with a 13-liter spirometer-rebreathing system during the hypoxic and hypercapnic tests). During room air breathing at 3,658 m (inspired O2 pressure = 93 Torr), Han-Tibetans resembled Tibetans in ventilation (12.1 +/- 0.6 vs. 11.5+/- 0.5 l/min BTPS, respectively) but had HVR that were blunted (63 +/- 16 vs. 121 +/- 13, respectively, for HVR shape parameter A) and declined with increasing duration of high-altitude residence. During administered hyperoxia (inspired O2 pressure = 310 Torr) at 3,658 m, the paradoxical hyperventilation previously seen in Tibetan but not Han residents at 3,658 m (11.8 +/- 0.5 vs. 10.1 +/- 0.5 l/min BTPS) was absent in these Han-Tibetans (9.8 +/- 0.6 l/min BTPS). Thus, although longer duration of high-altitude residence appears to progressively blunt HVR among Han-Tibetans born and residing at 3, 658 m, their Tibetan ancestry appears protective in their maintenance of high resting ventilation levels despite diminished chemosensitivity. PMID- 9390988 TI - Resistance training and human cervical muscle recruitment plasticity. AB - This study examined cervical neuromuscular adaptations to resistance training. The ResX group performed conventional resistance training plus head-extension exercise. Another group performed only conventional resistance training, and the control group performed no resistance exercise. Muscle use during head extension was determined by quantifying shifts in T2 in serial-transaxial magnetic resonance images of the neck. ResX was the only group that showed a training effect. Training decreased (P < 0.05) the cross-sectional area (CSA) of cervical muscle used to perform submaximal head extension by 31%. This reflected a decrease (P < 0.05) in relative use of the splenius capitis, semispinalis capitis, and semispinalis cervicis and multifidus muscles by about one-third; their percentage of CSA showing contrast shift was reduced from 60 to 40% on average. This same exercise evoked no contrast shift in the levator scapulae, longissimus capitis and cervicis, and scalenus medius and anterior muscles posttraining, yet 20% or more of their CSA was engaged pretraining. The relative CSA of cervical musculature that was used to perform maximal head extension was increased (P < 0.05) 16% by training. The findings suggest functional redundancy of neck musculature that can be modified by training; submaximal tasks can be performed despite cessation of recruitment of individual muscles, yet recruitment can be increased for maximal efforts. These results also suggest that neuromuscular adaptations to training require a specific cervical exercise. PMID- 9390989 TI - Improving energy expenditure estimation by using a triaxial accelerometer. AB - In our study of 125 subjects (53 men and 72 women) for two 24-h periods, we validated energy expenditure (EE), estimated by a triaxial accelerometer (Tritrac R3D), by using a whole-room indirect calorimeter under close-to-normal living conditions. The estimated EE was correlated with the measured total EE for the 2 days (r = 0. 925 and r = 0.855; P < 0.001) and in minute-by-minute EE (P < 0.01). Resting EE formulated by the Tritrac was found to be similar to the measured values [standard errors of estimation (SEE) = 0.112 W/kg; P = 0.822]. The Tritrac significantly underestimated total EE, EE for physical activities, EE of sedentary and light-intensity activities, and EE for exercise such as stepping (all P < 0.001). We developed a linear and a nonlinear model to predict EE by using the acceleration components from the Tritrac. Predicted EE was significantly improved with both models in estimating total EE, total EE for physical activities, EE in low-intensity activities, minute-by-minute averaged relative difference, and minute-by-minute SEE (all P < 0. 05). Furthermore, with our generalized models and by using subjects' physical characteristics and body acceleration, EE can be estimated with higher accuracy (averaged SEE = 0.418 W/kg) than with the Tritrac model. PMID- 9390990 TI - Modulation of pulmonary arterial input impedance during transition from inspiration to expiration. AB - We investigated whether respiration influences pulmonary arterial input impedance during transition from inspiration to expiration in five anesthetized, spontaneously breathing dogs. Impedance (Z) was separately assessed for heart beats occurring in inspiration, in expiration, and during the transition from inspiration to expiration (transitional beat). Transitional beats were scored by the ratio between the fraction of beat falling in expiration and the total beat duration [expiratory fraction (Efr)] to quantify their position within the transition. In transitional beats, input resistance linearly increased with Efr; Z modulus at the heart-rate frequency (fHR) decreased up to -50% for Efr = 50%. Z phase at fHR was greater than in inspiration for Efr <40% and lower for Efr >50%. Unlike blood flow velocity, mean value and first harmonic of pulmonary arterial pressure were correlated to Efr and paralleled the changes of input resistance and Z at fHR. This indicates that respiration influences Z through modifications in arterial pressure. The evidence of important respiratory influences on Z function may help the pathophysiological interpretation of dysfunctions of the right heart pumping action, such as the so-called cor pulmonale. PMID- 9390991 TI - Posttetanic potentiation of human dorsiflexors. AB - Twitch contractions of the ankle dorsiflexors were evoked before and after applied 7-s tetanic stimulation at 100 Hz in 20 young adults. Torque decreased 15% during the tetanus. At 5 s after tetanus, twitch peak torque had potentiated 45%. Potentiation declined to 28% after 1 min, rose slightly to 33% at 2 min, and declined slowly with potentiation still 25% after 5 min. There was large intersubject variation in the amount of potentiation (5-140%) and its persistence (5 to >/=20 min). The muscle compound action potential (M wave) did not change significantly (from pretetanic value) at 5 s after tetanus but increased sharply (26%) at 2 min and then subsided. Twitch half relaxation time (23%) decreased significantly more than twitch rise time (13%) 5 s after tetanus and recovered more slowly. Twitch rates of torque development (75%) and relaxation (71%) increased similarly 5 s after tetanus and were still elevated (approximately 25%) at 5 min. The extent of twitch torque potentiation was significantly inversely correlated with pretetanic twitch rise time (r = -0.69), half relaxation time (r = -0.61), and twitch-to-tetanus ratio (r = -0.66). The data indicate that posttetanic potentiation has a greater effect on twitch half relaxation time than on time to peak torque and is more prominent in muscles with a short twitch time course and small twitch-to-tetanus ratio. PMID- 9390992 TI - Effects of head-down-tilt bed rest on cerebral hemodynamics during orthostatic stress. AB - Our aim was to determine whether the adaptation to simulated microgravity (microG) impairs regulation of cerebral blood flow (CBF) during orthostatic stress and contributes to orthostatic intolerance. Twelve healthy subjects (aged 24 +/- 5 yr) underwent 2 wk of -6 degrees head-down-tilt (HDT) bed rest to simulate hemodynamic changes that occur when humans are exposed to microG. CBF velocity in the middle cerebral artery (transcranial Doppler), blood pressure, cardiac output (acetylene rebreathing), and forearm blood flow were measured at each level of a ramped protocol of lower body negative pressure (LBNP; -15, -30, and -40 mmHg x 5 min, -50 mmHg x 3 min, then -10 mmHg every 3 min to presyncope) before and after bed rest. Orthostatic tolerance was assessed by using the cumulative stress index (CSI; mmHg x minutes) for the LBNP protocol. After bed rest, each individual's orthostatic tolerance was reduced, with the group CSI decreased by 24% associated with greater decreases in cardiac output and greater increases in systemic vascular resistance at each level of LBNP. Before bed rest, mean CBF velocity decreased by 14, 10, and 45% at -40 mmHg, -50 mmHg, and maximal LBNP, respectively. After bed rest, mean velocity decreased by 16% at -30 mmHg and by 21, 35, and 39% at -40 mmHg, -50 mmHg, and maximal LBNP, respectively. Compared with pre-bed rest, post-bed-rest mean velocity was less by 11, 10, and 21% at -30, -40, and -50 mmHg, respectively. However, there was no significant difference at maximal LBNP. We conclude that cerebral autoregulation during orthostatic stress is impaired by adaptation to simulated microG as evidenced by an earlier and greater fall in CBF velocity during LBNP. We speculate that impairment of cerebral autoregulation may contribute to the reduced orthostatic tolerance after bed rest. PMID- 9390993 TI - An improved statistical methodology to estimate and analyze impedances and transfer functions. AB - Estimating the mathematical relationship between pulsatile time series (e.g., pressure and flow) is an effective technique for studying dynamic systems. The frequency-domain relationship between time series, often calculated as an impedance (pressure/flow), is known more generally as a frequency-response or transfer function (output/input). Current statistical methods for transfer function analysis 1) assume erroneously that repeated observations on a subject are independent, 2) have limited statistical value and power, or 3) are restricted to use in single subjects rather than in an entire sample. This paper develops a regression model for transfer function analysis that corrects each of these deficiencies. Spectral densities of the input and output time series and the cross-spectral density between them are first estimated from discrete Fourier transforms and then used to obtain regression estimates of the transfer function. Statistical comparisons of the transfer function estimates use a test statistic that is distributed as chi2. Confidence intervals for amplitude and phase can also be calculated. By correctly modeling repeated observations on each subject, this improved statistical approach to transfer function estimation and analysis permits the simultaneous analysis of data from all subjects in a sample, improves the power of the transfer function model, and has broad relevance to the study of dynamic physiological systems. PMID- 9390994 TI - Allometric modeling does not determine a dimensionless power function ratio for maximal muscular function. AB - In the exercise sciences, simple allometry (y = axb) is rapidly becoming the method of choice for scaling physiological and human performance data for differences in body size. The purpose of this study is to detail the specific regression diagnostics required to validate such models. The sum (T, in kg) of the "snatch" and "clean-and-jerk" lifts of the medalists from the 1995 Men's and Women's World Weightlifting Championships was modeled as a function of body mass (M, in kg). A log-linearized allometric model (ln T = ln a + b ln M) yielded a common mass exponent (b) of 0. 47 (95% confidence interval = 0.43-0.51, P < 0.01). However, size-related patterned deviations in the residuals were evident, indicating that the allometric model was poorly specified and that the mass exponent was not size independent. Model respecification revealed that second order polynomials provided the best fit, supporting previous modeling of weightlifting data (R. G. Sinclair. Can. J. Appl. Sport Sci. 10: 94-98, 1985). The model parameters (means +/- SE) were T = (21.48 +/- 16.55) + (6.119 +/- 0.359)M - (0. 022 +/- 0.002)M2 (R2 = 0.97) for men and T = (-20.73 +/- 24.14) + (5. 662 +/- 0.722)M - (0.031 +/- 0.005)M2 (R2 = 0.92) for women. We conclude that allometric scaling should be applied only when all underlying model assumptions have been rigorously evaluated. PMID- 9390995 TI - Differential dependence on GluR2 expression of three characteristic features of AMPA receptors. AB - The GluR2 subunit controls three key features of ion flux through the AMPA subtype of glutamate receptors-calcium permeability, inward rectification, and channel block by external polyamines, but whether each of these features is equally sensitive to GluR2 abundance is unknown. The relations among these properties were compared in native AMPA receptors expressed by acutely isolated hippocampal interneurons and in recombinant receptors expressed by Xenopus oocytes. The shape of current-voltage (I-V) relations between -100 and +50 mV for either recombinant or native AMPA receptors was well described by a Woodhull block model in which the affinity for internal polyamine varied over a 1000-fold range in different cells. In oocytes injected with mixtures of GluR2:non-GluR2 mRNA, the relative abundance of GluR2 required to reduce the log of internal blocker affinity by 50% was two- to fourfold higher than that needed to half maximally reduce divalent permeability or channel block by external polyamines. Likewise, in interneurons the affinity of externally applied argiotoxin for its blocking site was a steep function of internal blocker affinity. These results indicate that the number of GluR2 subunits in AMPA receptors is variable in both oocytes and interneurons. More GluR2 subunits in an AMPA receptor are required to maximally reduce internal blocker affinity than to abolish calcium permeability or external polyamine channel block. Accordingly, single-cell RT-PCR showed that approximately one-half of the physiologically characterized interneurons exhibiting inwardly rectifying AMPA receptors expressed detectable levels of edited GluR2. The physiological effects of a moderate change in GluR2 relative abundance, such as occurs after ischemia or seizures or after chronic exposure to morphine, thus will be dependent on the ambient GluR2 level in a cell-specific manner. PMID- 9390996 TI - The beta-amyloid precursor protein of Alzheimer's disease enhances neuron viability and modulates neuronal polarity. AB - beta-Amyloid precursor protein (betaPP) can reside at neuron and glial cell surfaces or undergo proteolytic processing into secreted fragments. Although betaPP has been studied extensively, its precise physiological role is unknown. A line of transgenic knock-out mice selectively deficient in betaPP survive and breed but exhibit motor dysfunction and brain gliosis, consistent with a physiological role for betaPP in neuron development. To elucidate these functions, we cultured hippocampal neurons from wild-type and betaPP-deficient mice and compared their ability to attach, survive, and develop neurites. We found that hippocampal neurons from betaPP-deficient mice had diminished viability and retarded neurite development. We also compared the effects of betaPP secretory products, released from wild-type astrocytes, on process outgrowth from wild-type and betaPP-deficient hippocampal neurons. Outgrowth was enhanced at 1 d in the presence of wild-type astrocytes, as compared with betaPP deficient astrocytes. However, by 3 d, neurons had shorter axons but more minor processes with more branching when cocultured with wild-type astrocytes, as compared with betaPP-deficient astrocytes. Our data demonstrate that cell associated neuronal betaPP contributes to neuron viability, axonogenesis, and arborization and that betaPP secretory products modulate axon growth, dendrite branching, and dendrite numbers. PMID- 9390997 TI - Regulation of amyloid precursor protein catabolism involves the mitogen-activated protein kinase signal transduction pathway. AB - Catabolic processing of the amyloid precursor protein (APP) is subject to regulatory control by protein kinases. We hypothesized that this regulation involves sequential activation of the enzymes mitogen-activated protein kinase kinase (MEK) and extracellular signal-regulated protein kinase (ERK). In the present investigation, we provide evidence that MEK is critically involved in regulating APP processing by both nerve growth factor and phorbol esters. Western blot analysis of the soluble N-terminal APP derivative APPs demonstrated that the synthetic MEK inhibitor PD 98059 antagonized nerve growth factor stimulation of both APPs production and ERK activation in PC12 cells. Moreover, PD 98059 inhibited phorbol ester stimulation of APPs production and activation of ERK in both human embryonic kidney cells and cortical neurons. Furthermore, overexpression of a kinase-inactive MEK mutant inhibited phorbol ester stimulation of APP secretion and activation of ERK in human embryonic kidney cell lines. Most important, PD 98059 antagonized phorbol ester-mediated inhibition of Abeta secretion from cells overexpressing human APP695 carrying the "Swedish mutation." Taken together, these data indicate that MEK and ERK may be critically involved in protein kinase C and nerve growth factor regulation of APP processing. The mitogen-activated protein kinase cascade may provide a novel target for altering catabolic processing of APP. PMID- 9390998 TI - Identification of two nervous system-specific members of the erg potassium channel gene family. AB - Two new potassium channel genes, erg2 and erg3, that are expressed in the nervous system of the rat were identified. These two genes form a small gene family with the previously described erg1 (HERG) gene. The erg2 and erg3 genes are expressed exclusively in the nervous system, in marked contrast to erg1, which is expressed in both neural and non-neural tissues. All three genes are expressed in peripheral sympathetic ganglia. The erg3 channel produces a current that has a large transient component at positive potentials, whereas the other two channels are slowly activating delayed rectifiers. Expression of the erg1 gene in the sympathetic nervous system has potential implications for the etiology of the LQT2 form of the human genetic disease long QT syndrome. PMID- 9390999 TI - Crossed rhythmic synaptic input to motoneurons during selective activation of the contralateral spinal locomotor network. AB - To investigate the cellular mechanisms underlying locomotor-related left-right coordination, we monitored the crossed synaptic input to lumbar motoneurons during contralateral ventral root rhythmicity in the neonatal rat spinal cord in vitro. Using a longitudinal split-bath setup, one hemicord was kept in normal solution, whereas the contralateral hemicord was exposed to 5-HT and NMDA. With this approach, rhythmic bursting could be induced in the ventral roots on the agonist-exposed side, whereas the ventral roots on the agonist-free side remained silent. Intracellular recordings were made from L1-L3 motoneurons on the silent agonist-free side during rhythmic activity in the contralateral ventral roots. At the resting membrane potential, the typical crossed synaptic input was a rhythmic barrage of depolarizing IPSPs. This input modulated the frequency of spikes induced with depolarizing direct current by inhibiting firing in phase with the contralateral bursts. Intracellular chloride loading increased the amplitude of the IPSPs, suggesting that they were chloride-dependent. Strychnine but not bicuculline generally blocked the rhythmic inhibitory input when added to the agonist-free side during contralateral rhythmicity. APV and CNQX on the agonist free side abolished the rhythmic inhibitory input in most motoneurons but not in all. We suggest that rat spinal motoneurons receive a mainly glycinergic rhythmic inhibition from the contralateral half of the locomotor network. Unlike in simpler vertebrates, the crossed inhibition often appears to be at least disynaptic, involving inhibitory premotor neurons located on the same side as the receiving motoneurons. These premotor neurons are rhythmically excited via a crossed pathway that depends on glutamatergic transmission. PMID- 9391000 TI - Endogenous monocarboxylates sustain hippocampal synaptic function and morphological integrity during energy deprivation. AB - The ability to fuel neurons via glycogenolysis is believed to be an important function of glia. Indeed, the slow, rather than immediate, depression of synaptic transmission in hippocampal slices during exogenous glucose deprivation suggests that intrinsic energy reservoirs help to sustain neurotransmission. It is believed that glia fuel neighboring neurons via diffusible monocarboxylates such as pyruvate and lactate, although a role for glucose has been proposed also. Using alpha-cyano-4-hydroxycinnamate (4-CIN) to inhibit monocarboxylate transport and cytochalasin B (CCB) to inhibit glucose transport, we examined the role of glucose and monocarboxylates in supporting the functional and morphological integrity of hippocampal neurons during glucose deprivation. Although 200 microM 4-CIN failed to depress EPSPs supported by 10 mM glucose, pretreatment with 4-CIN accelerated the depression of EPSPs during glucose deprivation. 4-CIN also accelerated the decline in glucose-supported EPSPs after administration of 50 microM CCB, whereas CCB failed to alter the slow decay of pyruvate-supported EPSPs during pyruvate deprivation. 4-CIN did not alter the morphology of pyramidal neurons in the presence of 10 mM glucose but produced significant damage during glucose deprivation or CCB administration. These results suggest that endogenous monocarboxylates rather than glucose maintain neuronal integrity during energy deprivation. Furthermore, EPSPs supported by 2-3.3 mM glucose were sensitive to 4-CIN, suggesting that endogenous monocarboxylates are involved in maintaining neuronal function even under conditions of mild glucose deprivation. PMID- 9391001 TI - Importance of polysynaptic inputs and horizontal connectivity in the generation of tetanus-induced long-term potentiation in the rat auditory cortex. AB - Supragranular pyramidal neurons in the adult rat auditory cortex (AC) show marked long-term potentiation (LTP) of population spikes after tetanic white matter stimulation (TS). For determination of whether this marked LTP is specific to AC, LTP in rat AC slices was compared with LTP in slices of the visual cortex (VC). The amplitude of TS-induced LTP in AC was twice that in VC. LTP of EPSPs was also studied with perforated patch or whole-cell recording. Although the amplitude of TS-induced LTP of EPSPs in AC was larger that in VC, no cortical difference was found in LTP elicited by low-frequency stimulation paired with current injection. Neocortical LTP is dependent on the activation of NMDA receptors, and induction of LTP requires postsynaptic depolarization for removal of Mg2+ blockade of NMDA receptors. The postsynaptic depolarization elicited by TS in supragranular pyramidal neurons in AC was significantly larger than that in VC. Cutting of supragranular horizontal connections resulted in a decrease in the depolarization amplitude in AC but an increase in the depolarization amplitude in VC. The cortical difference in TS-induced LTP was diminished in the slices in which horizontal connections in supragranular layers were cut. The estimated density of horizontal axon collaterals of supragranular pyramidal neurons in AC was approximately twice that in VC. These results strongly suggest that the marked polysynaptic and postsynaptic depolarization during TS and the resulting marked LTP in AC are attributed to well developed horizontal axon collaterals of supragranular pyramidal neurons in AC. PMID- 9391002 TI - Study of proline-directed protein kinases involved in phosphorylation of the heavy neurofilament subunit. AB - The high-molecular-mass neurofilament subunit (NFH) is normally hypophosphorylated in the neuronal perikaryon and undergoes extensive phosphorylation after entering the initial axon segment. Aberrant hyperphosphorylation of perikaryal NFH is a common feature of many neurological diseases. In a previous study (), we demonstrated a correlation between phosphorylation of perikaryal NFH and induction of stress-activated protein kinase (SAPK)-gamma. In this report, we present direct evidence showing that the in vivo activation of SAPKs by an upstream activator (MEKK-1) caused extensive NFH phosphorylation. We also show that stress-activated p38 kinases were not involved in the phosphorylation of perikaryal NFH in cultured dorsal root ganglion neurons and that this process was reversible. SAPKgamma was shown to be located in both the cell body and the neurites of the cultured neurons, suggesting that it is likely to be involved in the phosphorylation of cytoplasmic substrates. These could include neuritic NFH, which is highly phosphorylated despite the demonstrated lack of cyclin-dependent kinase-5 activity in these neurons. Neuritic NFH was also highly phosphorylated in neuronal cultures devoid of Schwann cells, indicating that this form of post-translational modification does not require cues stemming from Schwann cell-axon contacts. Collectively, these findings provide significant new insights into mechanisms involved in NFH phosphorylation in normal neurons and in disease states characterized by aberrant phosphorylation of neurofilaments. PMID- 9391003 TI - Modulation of hypothalamic-pituitary-adrenal function by transgenic expression of interleukin-6 in the CNS of mice. AB - Interleukin-6 (IL-6) and IL-6 receptor mRNA and protein have been reported in different brain regions under normal and pathophysiological conditions. Although much is known about the hypothalamic-pituitary-adrenal (HPA) axis stimulation after acute administration, less is known about the chronic effects of IL-6 on the function of the HPA axis. In the present study, we examined the function of the HPA axis in transgenic mice in which constitutive expression of IL-6 under the control of the glial fibrillary acidic protein (GFAP) promoter was targeted to astrocytes in the CNS. GFAP-IL6 mice heterozygous or homozygous for the IL-6 transgene had normal basal plasma corticosterone levels but, after restraint stress, showed abnormally increased levels in a gene dose-dependent manner. The increased plasma corticosterone levels in the IL-6 transgenic mice were associated with increased adrenal corticosterone content and hyperplasia of both adrenal cortex and medulla. Notably, plasma adrenocorticotrophic hormone (ACTH) levels and pituitary ACTH content were either not changed or decreased in these mice, whereas plasma arginine vasopressin (AVP) was increased, supporting a role for AVP in response to acute immobilization stress. The reduced ACTH response together with the adrenal hyperplasia in the IL-6 transgenic mice suggests direct activation at the level of the adrenal gland that may be directly activated by AVP or sensitized to ACTH. A similar mechanism may play a role in the blunted ACTH response and elevated corticosterone levels under pathophysiological conditions observed in humans with high brain levels of IL-6. PMID- 9391004 TI - Luteinizing hormone-releasing hormone (LHRH) neurons maintained in hypothalamic slice explant cultures exhibit a rapid LHRH mRNA turnover rate. AB - Evidence indicates that neuropeptide gene expression is tightly coupled to biosynthesis and secretion. Moreover, rhythmic gene expression often accompanies rhythmic secretion. Luteinizing hormone-releasing hormone (LHRH) neurosecretion, which regulates gonadal function, is pulsatile, with interpulse intervals of approximately 1 hr and pulse decays of <30 min in rats. As a basis for a rapid fall in peptide secretion, we hypothesize that LHRH mRNA levels rapidly decay. To address this hypothesis, we examined LHRH mRNA turnover in primary postnatal LHRH neurons maintained in long-term hypothalamic/preoptic area slice explant cultures, using in situ hybridization histochemistry (ISHH). Relative LHRH mRNA content per cell was quantitated by single-cell analysis after transcription inhibition with 5, 6-dichloro-1-D-ribofuranosyl-benzimidazole (DRB) or actinomycin D. Cultures were maintained in serum-free medium with tetrodotoxin to suppress spontaneous electrical activity and hence assess only intrinsic cellular activity. A plot of LHRH mRNA level per cell versus DRB treatment time showed a rapid initial decay of LHRH mRNA (t1/2, 5-13 min), followed by a slower decay rate (t1/2, 329-344 hr). LHRH cell number after drug treatment as determined by immunocytochemistry did not change. Comparison of mammalian LHRH mRNA 3' untranslated regions showed two conserved regions. These data indicate that, in primary LHRH neurons, LHRH mRNA has an intrinsically high rate of turnover and a mRNA stabilization component. Foremost, decay of LHRH mRNA, the fastest reported for a neuropeptide to date, corresponds to the decay of LHRH peptide pulses. PMID- 9391006 TI - Myosin VIIA is required for aminoglycoside accumulation in cochlear hair cells. AB - Myosin VIIA is expressed by sensory hair cells and has a primary structure predicting a role in membrane trafficking and turnover, processes that may underlie the susceptibility of hair cells to aminoglycoside antibiotics. [3H]Gentamicin accumulation and the effects of aminoglycosides were therefore examined in cochlear cultures of mice with different missense mutations in the myosin VIIA gene, Myo7a, to see whether myosin VIIA plays a role in aminoglycoside ototoxicity. Hair cells from homozygous mutant Myo7ash1 mice, with a mutation in a nonconserved region of the myosin VIIA head, respond rapidly to aminoglycoside treatment and accumulate high levels of gentamicin. Hair cells from homozygous mutant Myo7a6J mice, with a mutation at a highly conserved residue close to the ATP binding site of the myosin VIIA head, do not accumulate [3H]gentamicin and are protected from aminoglycoside ototoxicity. Hair cells from heterozygotes of both alleles accumulate [3H]gentamicin and respond to aminoglycosides. Although aminoglycoside uptake is thought to be via apical surface-associated endocytosis, coated pit numbers on the apical membrane of heterozygous and homozygous Myo7a6J hair cells are similar. Pulse-chase experiments with cationic ferritin confirm that the apical endocytotic pathway is functional in homozygous Myo7a6J hair cells. Transduction currents can be recorded from both heterozygous and homozygous Myo7a6J hair cells, suggesting it is unlikely that the drug enters via diffusion through the mechanotransducer channel. The results show that myosin VIIA is required for aminoglycoside accumulation in hair cells. Myosin VIIA may transport a putative aminoglycoside receptor to the hair cell surface, indirectly translocate it to sites of membrane retrieval, or retain it in the endocytotic pathway. PMID- 9391005 TI - Activity-dependent dendritic targeting of BDNF and TrkB mRNAs in hippocampal neurons. AB - The mechanisms underlying the subcellular localization of neurotrophins and their receptors are poorly understood. We show that in cultured hippocampal neurons, the mRNAs for BDNF and TrkB have a somatodendritic localization, and we quantify the extent of their dendritic mRNA localization. In the dendrites the labeling covers on average the proximal 30% of the total dendritic length. On high potassium depolarization, the labeling of BDNF and TrkB mRNA extends on average to 68% of the dendritic length. This increase does not depend on new RNA synthesis, is inhibited by the Na+ channel blocker tetrodotoxin, and involves the activation of glutamate receptors. Extracellular Ca2+, partly flowing through L type Ca2+ channels, is absolutely required for this process to occur. At the protein level, a brief stimulation of hippocampal neurons with 10 mM KCl leads to a marked increase of BDNF and TrkB immunofluorescence density in the distal portion of dendrites, which also occurs, even if at lower levels, when transport is inhibited by nocodazole. The protein synthesis inhibitor cycloheximide abolishes this increase. The activity-dependent modulation of mRNA targeting and protein accumulation in the dendrites may provide a mechanism for achieving a selective local regulation of the activity of neurotrophins and their receptors, close to their sites of action. PMID- 9391007 TI - Identification of caveolin and caveolin-related proteins in the brain. AB - Caveolae are 50-100 nm, nonclathrin-coated, flask-shaped plasma membrane microdomains that have been identified in most mammalian cell types, except lymphocytes and neurons. To date, multiple functions have been ascribed to caveolae, including the compartmentalization of lipid and protein components that function in transmembrane signaling events, biosynthetic transport functions, endocytosis, potocytosis, and transcytosis. Caveolin, a 21-24 kDa integral membrane protein, is the principal structural component of caveolae. We have initiated studies to examine the relationship of detergent-insoluble complexes identified in astrocytes to the caveolin-caveolae compartment detected in cells of peripheral tissues. Immunolocalization studies performed in astrocytes reveal caveolin immunoreactivity in regions that correlate well to the distribution of caveolae and caveolin determined in other cell types, and electron microscopic studies reveal multiple clusters of flask-shaped invaginations aligned along the plasma membrane. Immunoblot analyses demonstrate that detergent-insoluble complexes isolated from astrocytes are composed of caveolin-1alpha, an identification verified by Northern blot analyses and by the cloning of a cDNA using reverse transcriptase-PCR amplification from total astrocyte RNA. Using a full-length caveolin-1 probe, Northern blot analyses suggest that the expression of caveolin-1 may be regulated during brain development. Immunoblot analyses of detergent-insoluble complexes isolated from cerebral cortex and cerebellum identify two immunoreactive polypeptides with apparent molecular weight and isoelectric points appropriate for caveolin. The identification of caveolae microdomains and caveolin-1 in astrocytes and brain, as well as the apparent regulation of caveolin-1 expression during brain development, identifies a cell compartment not detected previously in brain. PMID- 9391008 TI - Sublethal oxygen-glucose deprivation alters hippocampal neuronal AMPA receptor expression and vulnerability to kainate-induced death. AB - Recent studies have suggested that rats subjected to transient global brain ischemia develop depressed expression of GluR-B in CA1 hippocampal neurons. The present study was performed to determine whether a similar change in AMPA receptor expression could be triggered in vitro by sublethal oxygen-glucose deprivation in rat hippocampal neuronal cultures. mRNA was extracted from individual hippocampal neurons via patch electrodes and amplified by RT-PCR 24-48 hr after sublethal oxygen-glucose deprivation. Compared with controls, insulted neurons expressed increased levels of GluR-D flop. As an indication that this change in receptor expression was functionally significant, insulted cultures exhibited increased AMPA- or kainate-induced 45Ca2+ accumulation sensitive to Joro spider toxin and increased vulnerability to kainate-induced death. These data support the hypothesis that exposure to ischemia may enhance subsequent hippocampal neuronal vulnerability to AMPA receptor-mediated excitotoxicity by modifying the relative expression of AMPA receptor subunits in a manner that promotes Ca2+ permeability. PMID- 9391009 TI - Repression of the calcitonin gene-related peptide promoter by 5-HT1 receptor activation. AB - We have investigated the control of calcitonin gene-related peptide (CGRP) expression by a serotonergic agonist that is related pharmacologically to currently used antimigraine drugs. During migraines, CGRP levels are elevated but then returned to normal by a 5-HT1 receptor agonist, sumatriptan. However, neither the molecular nor cellular targets of this drug are known. Trigeminal neurons are the major source of cerebrovascular CGRP, and thus we have used trigeminal primary cultures and the neuronal-like CA77 thyroid C-cell line as a model. We first demonstrate that sumatriptan and another 5-HT1 agonist, CGS 12066A (CGS), cause a robust and prolonged increase with oscillations in intracellular calcium in CA77 cells. CGS caused a similar increase in trigeminal cultures. We then show that CGS treatment leads to a decrease in CGRP mRNA levels in the CA77 cells. This decrease is attributable to the repression of promoter activity through two discrete elements: (1) the cAMP-responsive region, via a cAMP-independent mechanism; and (2) the cell-specific enhancer, which binds the upstream stimulatory factor helix-loop-helix protein and a cell-specific activator. These results demonstrate that activation of the endogenous 5-HT1 receptor is coupled to calcium signaling pathways and leads to inhibition of CGRP gene transcription. PMID- 9391010 TI - Measurement of intracellular free zinc in living cortical neurons: routes of entry. AB - We used the ratioable fluorescent dye mag-fura-5 to measure intracellular free Zn2+ ([Zn2+]i) in cultured neocortical neurons exposed to neurotoxic concentrations of Zn2+ in concert with depolarization or glutamate receptor activation and identified four routes of Zn2+ entry. Neurons exposed to extracellular Zn2+ plus high K+ responded with a peak cell body signal corresponding to a [Zn2+]i of 35-45 nM. This increase in [Zn2+]i was attenuated by concurrent addition of Gd3+, verapamil, omega-conotoxin GVIA, or nimodipine, consistent with Zn2+ entry through voltage-gated Ca2+channels. Furthermore, under conditions favoring reverse operation of the Na+-Ca2+ exchanger, Zn2+ application induced a slow increase in [Zn2+]i and outward whole-cell current sensitive to benzamil-amiloride. Thus, a second route of Zn2+ entry into neurons may be via transporter-mediated exchange with intracellular Na+. Both NMDA and kainate also induced rapid increases in neuronal [Zn2+]i. The NMDA-induced increase was only partly sensitive to Gd3+ or to removal of extracellular Na+, consistent with a third route of entry directly through NMDA receptor-gated channels. The kainate induced increase was highly sensitive to Gd3+ or Na+ removal in most neurons but insensitive in a minority subpopulation ("cobalt-positive cells"), suggesting that a fourth route of neuronal Zn2+ entry is through the Ca2+-permeable channels gated by certain subtypes of AMPA or kainate receptors. PMID- 9391011 TI - Isoform specificity in the relationship of actin to dendritic spines. AB - Dendritic spines contain high concentrations of actin, but neither the isoforms involved nor the mechanism of accumulation is known. In situ hybridization with specific probes established that beta- and gamma-cytoplasmic actins are selectively expressed at high levels by spine-bearing neurons. Transfecting cultured hippocampal neurons with epitope-tagged actin isoforms showed that cytoplasmic beta- and gamma-cytoplasmic actins are correctly targeted to spines, whereas alpha-cardiac muscle actin, which is normally absent from neurons, formed aggregates in dendrites. The transfected actin cDNAs contained only coding domains, suggesting that spine targeting involves amino acid sequences in the proteins, an interpretation supported by experiments with chimeric cDNAs in which C-terminal actin sequences were found to be determinative in spine targeting. By contrast to actin, microtubule components, including tubulin and MAP2, were restricted to the dendritic shaft domain. The close association of cytoplasmic actins with spines together with their general involvement in cell surface motility further supports the idea that actin motility-based changes in spine shape may contribute to synaptic plasticity. PMID- 9391012 TI - Nerve growth factor modulates synaptic transmission between sympathetic neurons and cardiac myocytes. AB - Regulation of heart rate by the sympathetic nervous system involves the release of norepinephrine (NE) from nerve terminals onto heart tissue, resulting in an elevation in beat rate. Nerve growth factor (NGF) is a neurotrophin produced by the heart that supports the survival and differentiation of sympathetic neurons. Here we report that NGF also functions as a modulator of sympathetic synaptic transmission. We determined the effect of NGF on the strength of synaptic transmission in co-cultures of neonatal rat cardiac myocytes and sympathetic neurons from the superior cervical ganglion (SCG). Synaptic transmission was assayed functionally, as an increase in the beat rate of a cardiac myocyte during stimulation of a connected neuron. Application of NGF produced a pronounced, reversible enhancement of synaptic strength. We found that TrkA, the receptor tyrosine kinase that mediates many NGF responses, is expressed primarily by neurons in these cultures, suggesting a presynaptic mechanism for the effects of NGF. A presynaptic model is further supported by the finding that NGF did not alter the response of myocytes to application of NE. In addition to the acute modulatory effects of NGF, we found that the concentration of NGF in the growth medium affects the level of synaptic transmission in cultures of sympathetic neurons and cardiac myocytes. These results indicate that in addition to its role as a survival factor, NGF plays both acute and long-term roles in the regulation of developing sympathetic synapses in the cardiac system. PMID- 9391014 TI - Experience-dependent modifications in MAP2 phosphorylation in rat olfactory bulb. AB - Microtubule-associated protein 2 (MAP2) is a neuron-specific cytoskeletal protein, enriched in dendrites and cell bodies, that helps determine dendritic shape. MAP2 regulates microtubule stability in a phosphorylation-dependent manner. The present study used immunocytochemistry with phosphoepitope-specific and phosphorylation state-independent antibodies to examine experience-dependent changes in MAP2 expression during postnatal development of the olfactory bulb. Our results demonstrate that immunoreactivity reflecting total MAP2 expression reaches a maximal level by postnatal day 20 (P20). The degree of staining for phosphoindependent forms of MAP2 is relatively unaffected by blocking odorant passage to one half the nasal epithelium via unilateral naris closure, a manipulation that attenuates physiological activity in the bulb. However, olfactory restriction from P1 dramatically reduces immunoreactivity for antibody AP18, which recognizes MAP2 only when phosphorylated on Ser136. Quantification of staining in the granule cell layer indicates that the greatest difference (64%) between control and experimental bulbs occurs after occlusion from P1 to P30 compared with animals deprived from P1 to P10 or P1 to P20. The shift in MAP2 phosphorylation occurs even when deprivation is delayed until P30, after the sensitive period for experience-dependent changes in bulb volume. Thus, the degree of the phosphorylation shift depends on the duration but not the time of onset of naris closure. Because staining for phosphorylation-independent forms of MAP2 is unchanged by naris closure, the total amount of the protein per unit area is probably not significantly altered. However, the large reductions of AP18 immunoreactivity in the bulb after olfactory restriction suggest that there is an activity-dependent stimulation of MAP2 phosphorylation. PMID- 9391013 TI - BDNF and NT-4/5 prevent atrophy of rat rubrospinal neurons after cervical axotomy, stimulate GAP-43 and Talpha1-tubulin mRNA expression, and promote axonal regeneration. AB - Rubrospinal neurons (RSNs) undergo a marked atrophy in the second week after cervical axotomy. This delayed atrophy is accompanied by a decline in the expression of regeneration-associated genes such as GAP-43 and Talpha1-tubulin, which are initially elevated after injury. These responses may reflect a deficiency in the trophic support of axotomized RSNs. To test this hypothesis, we first analyzed the expression of mRNAs encoding the trk family of neurotrophin receptors. In situ hybridization revealed expression of full-length trkB receptors in virtually all RSNs, which declined 7 d after axotomy. Full-length trkC mRNA was expressed at low levels. Using RT-PCR, we found that mRNAs encoding trkC isoforms with kinase domain inserts were present at levels comparable to that for the unmodified receptor. TrkA mRNA expression was not detected in RSNs, and the expression of p75 was restricted to a small subpopulation of axotomized cells. In agreement with the pattern of trk receptor expression, infusion of recombinant human BDNF or NT-4/5 into the vicinity of the axotomized RSNs, between days 7 and 14 after axotomy, fully prevented their atrophy. This effect was still evident 2 weeks after the termination of BDNF treatment. Moreover, BDNF or NT-4/5 treatment stimulated the expression of GAP-43 and Talpha1-tubulin mRNA and maintained the level of trkB expression. Vehicle, NGF, or NT-3 treatment had no significant effect on cell size or GAP-43 and Talpha1-tubulin expression. In a separate experiment, infusion of BDNF also was found to increase the number of axotomized RSNs that regenerated into a peripheral nerve graft. Thus, in BDNF treated animals, the prevention of neuronal atrophy and the stimulation GAP-43 and Talpha1-tubulin expression is correlated with an increased regenerative capacity of axotomized RSNs. PMID- 9391015 TI - Sexual dimorphism in the spinal cord is absent in mice lacking the ciliary neurotrophic factor receptor. AB - Ciliary neurotrophic factor (CNTF) has potent survival-promoting effects on motoneurons in vitro and in vivo. We examined knockout mice with null mutations of the gene for either CNTF itself or the alpha-subunit of the CNTF receptor (CNTFRalpha) to assess whether CNTF and/or its receptors are involved in the development of a sexually dimorphic neuromuscular system. Male rodents have many more motoneurons in the spinal nucleus of the bulbocavernosus (SNB) than do females. This sex difference is caused by hormone-regulated death of SNB motoneurons and their target muscles. Sexual dimorphism of SNB motoneuron number developed completely normally in CNTF knockout (CNTF -/-) mice. In contrast, a sex difference in the SNB was absent in CNTFRalpha -/- animals: male mice lacking a functional CNTF alpha-receptor had fewer than half as many SNB motoneurons than did wild-type males and no more than did their female counterparts. Size of the bulbocavernosus and levator ani muscles, the main targets of SNB motoneurons, was not affected in either CNTF or CNTFRalpha knockout males. These observations suggest that signaling through the CNTF receptor is involved in sexually dimorphic development of SNB motoneuron number and that target muscle survival per se is not sufficient to ensure motoneuron survival in this system. In addition, our observations are consistent with the suggestion that CNTF itself is not the only endogenous ligand for the CNTF receptor. A second, as yet unknown, ligand may be important for neural development, including sexually dimorphic motoneuron development. PMID- 9391016 TI - Impaired parallel fiber-->Purkinje cell synapse stabilization during cerebellar development of mutant mice lacking the glutamate receptor delta2 subunit. AB - The glutamate receptor delta2 subunit (GluRdelta2) is specifically expressed in cerebellar Purkinje cells (PCs) from early developmental stages and is selectively localized at dendritic spines forming synapses with parallel fibers (PFs). Targeted disruption of the GluRdelta2 gene leads to a significant reduction of PF-->PC synapses. To address its role in the synaptogenesis, the morphology and electrophysiology of PF-->PC synapses were comparatively examined in developing GluRdelta2 mutant and wild-type cerebella. PCs in GluRdelta2 mutant mice were normally produced, migrated, and formed spines, as did those in wild type mice. At the end of the first postnatal week, 74-78% of PC spines in both mice formed immature synapses, which were characterized by small synaptic contact, few synaptic vesicles, and incomplete surrounding by astroglial processes, eliciting little electrophysiological response. During the second and third postnatal weeks when spines and terminals are actively generated, the percentage of PC spines forming synapses attained 98-99% in wild type but remained as low as 55-60% in mutants, and the rest were unattached to any nerve terminals. As a result, the number of PF synapses per single-mutant PCs was reduced to nearly a half-level of wild-type PCs. Parallelly, PF stimulation less effectively elicited EPSCs in mutant PCs than in wild-type PCs during and after the second postnatal week. These results suggest that the GluRdelta2 is involved in the stabilization and strengthening of synaptic connectivity between PFs and PCs, leading to the association of all PC spines with PF terminals to form functionally mature synapses. PMID- 9391017 TI - Macrophage/Microglia regulation of astrocytic tenascin: synergistic action of transforming growth factor-beta and basic fibroblast growth factor. AB - After injury to the CNS, extracellular matrix molecules such as tenascin are upregulated around the injury site and may be involved in inhibition of axon growth. In the present study, astrocytes were investigated to determine which cell types, growth factors, or cytokines are responsible for the injury-induced regulation of tenascin. The addition of activated macrophage- or microglial conditioned medium increased astrocytic expression of tenascin 2.5-fold, as determined by Northern and Western blot analysis and ELISA. Of the cytokines and growth factors examined, only transforming growth factor-beta1 (TGF-beta1) and basic fibroblast growth factor (bFGF) significantly induced an increase in the production of astrocytic tenascin. Examination of macrophage and microglial supernatants showed the presence of TGF-beta1 but not bFGF; however, the TGF beta1 concentration in supernatants was lower than that expected to induce an increase in astrocytic tenascin similar to that seen with recombinant TGF-beta1. Western blot analysis of astrocytes showed only the presence of bFGF. Compared with the responses of the individual growth factors, tenascin production by astrocytes was dramatically potentiated when grown in the presence of a combination of both TGF-beta1 and bFGF. A similar synergistic effect was observed after the addition of either TGF-beta1 or bFGF to macrophage-conditioned medium. Northern analysis also showed concomitant increases in TGF-beta1, bFGF, and tenascin after CNS injury to animals 14 d of age or older. These results show that the regulation of astrocytic tenascin is mediated by the synergistic action of TGF-beta1 and bFGF in vitro and after injury in vivo. PMID- 9391018 TI - Development of membrane properties in taste cells of fungiform papillae: functional evidence for early presence of amiloride-sensitive sodium channels. AB - Behavioral and physiological studies have demonstrated a reduced sensitivity to several taste stimuli early in development. It has been suggested that this reduced sensitivity results from a late maturation of underlying transduction mechanisms. Little is known, however, about maturation of membrane properties of taste cells early in development. We have obtained whole-cell recordings from single fungiform taste cells of rat pups to examine the development of the NaCl transduction system. Although taste buds undergo a considerable increase in size during development, membrane capacitance measurements revealed no change in membrane surface area of individual taste cells, suggesting that the increase in size results from an increase in the total number of cells per bud. Whole-cell recordings showed that taste cells from very young pups [postnatal day 2 (PND2)] already possessed voltage-activated Na+ and K+ currents with no apparent differences in size or kinetics compared with adults. Surprisingly, amiloride sensitive Na+ responses, important for Na+ transduction, were found as early as PND2. The magnitude of responses to amiloride and the percentage of amiloride sensitive cells remained the same throughout all age groups. Furthermore, the similarity of amiloride inhibition constants suggested that the channel in neonates is the same channel that is expressed in adult taste buds. Our results indicate that taste cells at PND2 already have acquired the transduction elements necessary for signaling NaCl responses to the afferent nerve. We hypothesize that complete functionality of the salt taste transduction system, however, may not be reached until amiloride-sensitive Na+ channels become selectively localized at the apical membrane. This would explain previous studies indicating that amiloride sensitivity cannot be detected before PND12 in the intact tongue. Apical clustering of channels along with the opening of the taste pore and an increase in the total number of taste cells per bud likely constitute additional important steps toward a fully functional sensory system. PMID- 9391019 TI - The origin, location, and projections of the embryonic abdominal motorneurons of Drosophila. AB - We have used a retrograde labeling technique to identify motorneurons for each of the 30 body wall muscles of an abdominal hemisegment in the late stage 16 Drosophila embryo. Each motorneuron has a characteristic cell body position, dendritic arborization, and axonal projection. In addition, we have determined the neuroblasts of origin for most of the motorneurons we describe. Some organizational principles for the neuromuscular system have become apparent: (1) There is no obvious topographic relationship between the cell body positions of motorneurons and the position or orientation of the muscles they innervate; (2) motorneurons that innervate muscles of similar position and orientation are often clustered and have overlapping dendritic trees; (3) morphologically similar motorneurons are generally derived from a common neuroblast and innervate operationally related muscles; and (4) neuroblasts can give rise to more than one morphological type of motorneuron. PMID- 9391020 TI - Analysis of the mechanism of loss of trophic factor dependence associated with neuronal maturation: a phenotype indistinguishable from Bax deletion. AB - During development, sympathetic neurons are critically dependent on nerve growth factor (NGF) for survival. Neurons isolated from the superior cervical ganglia (SCG) of embryonic rodents and maintained for 1 week in vitro undergo programmed cell death in response to NGF deprivation. As the cells mature in vitro and in vivo, however, these neurons develop a resistance to NGF deprivation and become much less acutely dependent on NGF for survival. Using an in vitro model of neuronal maturation, we confirmed that SCG neurons maintained in culture for 3-4 weeks did not experience a dramatic loss in viability after NGF removal, yet they did undergo the initial biochemical and genetic changes elicited by NGF deprivation of young neurons. NGF deprivation of mature neurons produced rapid decreases in glucose uptake and protein and RNA synthesis rates, increased phosphorylation of c-Jun, and an increase in c-jun mRNA. Mature neurons, however, experienced a block in the cell death program before the final stages of the pathway activated in young neurons, which includes the induction of c-fos mRNA and characteristic apoptotic nuclear changes. This maturation-induced block was indistinguishable by these criteria from the block produced by Bax deficiency. Expression of Bax in mature neurons restored the apoptotic pathway, such that after NGF removal, Bax-overexpressing mature neurons resumed the apoptotic program, including the induction of c-Fos and passage through a caspase checkpoint. Thus, a block in the apoptotic program at or near the BAX checkpoint accounts for the decreased dependence of mature neurons on neurotrophic factor to maintain survival. PMID- 9391021 TI - Role of the supplementary motor area and the right premotor cortex in the coordination of bimanual finger movements. AB - To obtain a better understanding of the cortical representation of bimanual coordination, we measured regional cerebral blood flow (rCBF) with 15O-labeled water and positron emission tomography (PET). To detect areas with changes of rCBF during bimanual finger movements of different characteristics, we studied 12 right-handed normal volunteers. A complete session consisted of three rest scans and six scans with acoustically paced (1 Hz) bimanual, mirror, or parallel sequential finger movements. Activation of the right dorsal premotor area (PMd) extending to the posterior supplementary motor area (SMA) was significantly stronger during the parallel movements than during the mirror sequential movements (p < 0.05, at cluster level with correction for multiple comparisons). To determine whether these cortical areas truly represented bimanual coordination, a different group of nine normal volunteers was studied with a different task. Subjects performed acoustically paced (2 Hz) abduction-adduction movements of the index finger, making right only, left only, and bimanual mirror and parallel movements. Activation of the posterior SMA and right PMd was significantly greater during the parallel movements than during the bimanual mirror movements or the unimanual movements of either hand (p < 0.01, with anatomical constraint). Thus, the posterior SMA and right PMd appear to be related to the bimanual coordination of finger movements. PMID- 9391022 TI - Lobular patterns of cerebellar activation in verbal working-memory and finger tapping tasks as revealed by functional MRI. AB - The lobular distributions of functional activation of the cerebellum during verbal working-memory and finger movement tasks were investigated using functional magnetic resonance imaging (fMRI). Relative to a rest control, finger tapping of the right hand produced ipsilateral-increased activation in HIV/HV [Roman numeral designations based on Larsell's () nomenclature] and HVI and weaker activation in HVIII that was stronger on the ipsilateral side. For a working-memory task, subjects were asked to remember six (high load) or one (low load) visually presented letters across a brief delay. To assess the motoric aspects of rehearsal in the absence of working memory, we asked the subjects to repeatedly read subvocally six or one letters at a rate that approximated the internally generated rehearsal of working memory (motoric rehearsal task). For both tasks, bilateral regions of the superior cerebellar hemispheres (left superior HVIIA and right HVI) and portions of posterior vermis (VI and superior VIIA) exhibited increased activation during high relative to low load conditions. In contrast, the right inferior cerebellar hemisphere (HVIIB) exhibited this load effect only during the working-memory task. We hypothesize that HVI and superior HVIIA activation represents input from the articulatory control system of working memory from the frontal lobes and that HVIIB activation is derived from the phonological store in temporal and parietal regions. From these inputs, the cerebellum could compute the discrepancy between actual and intended phonological rehearsal and use this information to update a feedforward command to the frontal lobes, thereby facilitating the phonological loop. PMID- 9391023 TI - Insular cortical projections to functional regions of the striatum correlate with cortical cytoarchitectonic organization in the primate. AB - We examined the striatal projections from different cytoarchitectonic regions of the insular cortex using anterograde and retrograde techniques. The shell and medial ventral striatum receive inputs primarily from the agranular and ventral dysgranular insula. The central ventral striatum receives inputs primarily from the dorsal agranular and dysgranular insula. Projections to the central ventral striatum originate from more posterior and dorsal insular regions than projections to the medial ventral striatum. The dorsolateral striatum receives projections primarily from the dorsal dysgranular and granular insula. These results show that cytoarchitectonically less differentiated (agranular) insular regions project to the ventromedial "limbic" part of the ventral striatum, whereas more differentiated (granular) insular regions project to the dorsolateral "sensorimotor" part of the striatum. The finding that the ventral "limbic" striatum receives inputs from less differentiated regions of the insula is consistent with the general principle that less differentiated cortical regions project primarily to the "limbic" striatum. Functionally, the ventral striatum receives insular projections primarily related to integrating feeding behavior with rewards and memory, whereas the dorsolateral striatum receives insular inputs related to the somatosensation. Information regarding food acquisition in the insula may be sent to the intermediate area of the striatum. PMID- 9391024 TI - A neural model of multimodal adaptive saccadic eye movement control by superior colliculus. AB - How does the saccadic movement system select a target when visual, auditory, and planned movement commands differ? How do retinal, head-centered, and motor error coordinates interact during the selection process? Recent data on superior colliculus (SC) reveal a spreading wave of activation across buildup cells the peak activity of which covaries with the current gaze error. In contrast, the locus of peak activity remains constant at burst cells, whereas their activity level decays with residual gaze error. A neural model answers these questions and simulates burst and buildup responses in visual, overlap, memory, and gap tasks. The model also simulates data on multimodal enhancement and suppression of activity in the deeper SC layers and suggests a functional role for NMDA receptors in this region. In particular, the model suggests how auditory and planned saccadic target positions become aligned and compete with visually reactive target positions to select a movement command. For this to occur, a transformation between auditory and planned head-centered representations and a retinotopic target representation is learned. Burst cells in the model generate teaching signals to the spreading wave layer. Spreading waves are produced by corollary discharges that render planned and visually reactive targets dimensionally consistent and enable them to compete for attention to generate a movement command in motor error coordinates. The attentional selection process also helps to stabilize the map-learning process. The model functionally interprets cells in the superior colliculus, frontal eye field, parietal cortex, mesencephalic reticular formation, paramedian pontine reticular formation, and substantia nigra pars reticulata. PMID- 9391025 TI - The circumventricular organs form a potential neural pathway for lactate sensitivity: implications for panic disorder. AB - Patients with panic disorder experience panic attacks after intravenous sodium lactate infusions by an as yet unexplained mechanism. Lactate elicits a panic like response in rats with chronic dysfunction of GABA neurotransmission in the dorsomedial hypothalamus (DMH). The circumventricular organs, organum vasculosum lamina terminalis (OVLT) and subfornical organ (SFO), are potential sites that could detect increases in plasma lactate levels and activate the DMH. To test this, we obtained baseline heart rate (HR) and blood pressure (BP) responses to lactate infusions in rats fit with femoral arterial and venous catheters. Next, unilateral chronic injection cannulae connected to an Alzet infusion pump filled with the GABA synthesis inhibitor L-allylglycine (L-AG) were implanted into the DMH. Another chronic injection cannula was implanted into the region of the OVLT, SFO, or an adjacent control site, the median preoptic area (MePOA). These rats were tested once again with lactate infusions after injection of either artificial cerebrospinal fluid (ACSF) or tetrodotoxin (TTX) into the CVO sites. Injecting TTX into the OVLT completely blocked the lactate-induced response, whereas TTX injections into the SFO or MePOA did not. Also, direct injections of lactate (100 or 500 nl) into the OVLT elicited robust anxiety-like responses in these rats. These results suggest that the OVLT may be the primary site that detects lactate infusions, activating an anxiety-like response in a compromised DMH, and provide the first neuroanatomical basis for lactate response in panic disorder. PMID- 9391026 TI - The cerebellum and red nucleus are not required for In vitro classical conditioning of the turtle abducens nerve response. AB - The role of the cerebellum during motor learning is a controversial issue. Many authors have suggested that the cerebellum and its connections with the red nucleus are essential for the acquisition of the conditioned eye blink reflex. Although there is little argument that the cerebellum is an important component to the generation of the conditioned response (CR), a number of studies have suggested that the cerebellum is not essential for conditioning. Using an in vitro model of the classically conditioned turtle abducens nerve response, we investigated the effect of cerebellar and red nucleus lesions on the acquisition, extinction, and reacquisition of CRs. Neural discharge was recorded from the abducens nerve after a single shock unconditioned stimulus (US) was applied to the ipsilateral trigeminal nerve. When the US was paired with a conditioned stimulus (CS) applied to the posterior eighth, or auditory, nerve, a positive slope of CR acquisition was recorded in the abducens nerve. After extinction stimuli in which the CS and US were alternated, the number of CRs decreased to near zero. When the CS and US were once again paired, reacquisition at a faster rate was recorded. The CRs showed unusual timing features compared with preparations in which the cerebellum was intact; they had significantly shorter latencies and showed burst-like responses. These data demonstrate that it is possible to classically condition this in vitro preparation in the absence of the cerebellum and red nucleus. However, the latencies of CRs were found to be dramatically altered in the cerebellar-lesioned preparations, suggesting that the cerebellum does play a role in the timing of the CR. PMID- 9391027 TI - Effects of sleep on wake-induced c-fos expression. AB - We investigated the effects of sleep on wake-induced c-fos expression in the cerebral cortex of rats and c-fos-lacZ transgenic mice. In the cortex of rats, the levels of c-Fos, detected both by immunocytochemistry and Western blot, remained high during 6 or 12 hr of enforced wakefulness but declined rapidly (within 1 hr) with increasing time of recovery sleep. Similarly, in the transgenic mice in which lacZ expression is driven from the c-fos promoter, beta galactosidase activity was high after enforced wakefulness and declined with increasing amounts of sleep. These results suggest that the decrease in c-Fos protein in cortical neurons during sleep may be attributable to cessation of c fos expression, activation of a process that degrades the wake-induced c-Fos, or both. PMID- 9391029 TI - On the utility of nitrogen in leaves. PMID- 9391028 TI - A hypothesis about the endogenous analogue of general anesthesia. PMID- 9391030 TI - Weaving cartilage at zero g: the reality of tissue engineering in space. PMID- 9391031 TI - Ways and means for left shifts in the MaxiK channel. PMID- 9391032 TI - Metal ions and synaptic transmission: think zinc. PMID- 9391034 TI - Electric field-induced reorganization of two-component supported bilayer membranes. AB - Application of electric fields tangent to the plane of a confined patch of fluid bilayer membrane can create lateral concentration gradients of the lipids. A thermodynamic model of this steady-state behavior is developed for binary systems and tested with experiments in supported lipid bilayers. The model uses Flory's approximation for the entropy of mixing and allows for effects arising when the components have different molecular areas. In the special case of equal area molecules the concentration gradient reduces to a Fermi-Dirac distribution. The theory is extended to include effects from charged molecules in the membrane. Calculations show that surface charge on the supporting substrate substantially screens electrostatic interactions within the membrane. It also is shown that concentration profiles can be affected by other intermolecular interactions such as clustering. Qualitative agreement with this prediction is provided by comparing phosphatidylserine- and cardiolipin-containing membranes. PMID- 9391033 TI - Transcriptional activation: is it rocket science? PMID- 9391035 TI - A yeast genetic system for selecting small molecule inhibitors of protein-protein interactions in nanodroplets. AB - Cellular processes are mediated by complex networks of molecular interactions. Dissection of their role most commonly is achieved by using genetic mutations that alter, for example, protein-protein interactions. Small molecules that accomplish the same result would provide a powerful complement to the genetic approach, but it generally is believed that such molecules are rare. There are several natural products, however, that illustrate the feasibility of this approach. Split-pool synthesis now provides a simple mechanical means to prepare vast numbers of complex, even natural product-like, molecules individually attached to cell-sized polymer beads. Here, we describe a genetic system compatible with split-pool synthesis that allows the detection of cell-permeable, small molecule inhibitors of protein-protein interactions in 100- to 200-nl cell culture droplets, prepared by a recently described technique that arrays large numbers of such droplets. These "nanodroplets" contain defined media, cells, and one or more beads containing approximately 100 pmol of a photoreleasable small molecule and a controlled number of cells. The engineered Saccharomyces cerevisiae cells used in this study express two interacting proteins after induction with galactose whose interaction results in cell death in the presence of 5-fluoroorotic acid (inducible reverse two-hybrid assay). Disruption of the interaction by a small molecule allows growth, and the small molecule can be introduced into the system hours before induction of the toxic interaction. We demonstrate that the interaction between the activin receptor R1 and the immunophilin protein FKBP12 can be disrupted by the small molecule FK506 at nanomolar concentrations in nanodroplets. This system should provide a general method for selecting cell-permeable ligands that can be used to study the relevance of protein-protein interactions in living cells or organisms. PMID- 9391037 TI - Isotropic isotopy and symplectic null sets. AB - Capacity is an important numerical invariant of symplectic manifolds. This paper studies when a subset of a symplectic manifold is null, i.e., can be removed without affecting the ambient capacity. After examples of open null sets and codimension-2 non-null sets, geometric techniques are developed to perturb any isotopy of a loop to a hamiltonian flow; it follows that sets of dimension 0 and 1 are null. For isotropic sets of higher dimensions, obstructions to the perturbation are found in homotopy groups of the orthogonal groups. PMID- 9391036 TI - A nonnatural transcriptional coactivator. AB - In eukaryotes, sequence-specific DNA-binding proteins activate gene expression by recruiting the transcriptional apparatus and chromatin remodeling proteins to the promoter through protein-protein contacts. In many instances, the connection between DNA-binding proteins and the transcriptional apparatus is established through the intermediacy of adapter proteins known as coactivators. Here we describe synthetic molecules with low molecular weight that act as transcriptional coactivators. We demonstrate that a completely nonnatural activation domain in one such molecule is capable of stimulating transcription in vitro and in vivo. The present strategy provides a means of gaining external control over gene activation through intervention using small molecules. PMID- 9391038 TI - Characterization of residual structure in the thermally denatured state of barnase by simulation and experiment: description of the folding pathway. AB - Residual structure in the denatured state of a protein may contain clues about the early events in folding. We have simulated by molecular dynamics the denatured state of barnase, which has been studied by NMR spectroscopy. An ensemble of 10(4) structures was generated after 2 ns of unfolding and following for a further 2 ns. The ensemble was heterogeneous, but there was nonrandom, residual structure with persistent interactions. Helical structure in the C terminal portion of helix alpha1 (residues 13-17) and in helix alpha2 as well as a turn and nonnative hydrophobic clustering between beta3 and beta4 were observed, consistent with NMR data. In addition, there were tertiary contacts between residues in alpha1 and the C-terminal portion of the beta-sheet. The simulated structures allow the rudimentary NMR data to be fleshed out. The consistency between simulation and experiment inspires confidence in the methods. A description of the folding pathway of barnase from the denatured to the native state can be constructed by combining the simulation with experimental data from phi value analysis and NMR. PMID- 9391039 TI - Catalytic mechanism of the adenylyl and guanylyl cyclases: modeling and mutational analysis. AB - The adenylyl and guanylyl cyclases catalyze the formation of 3', 5'-cyclic adenosine or guanosine monophosphate from the corresponding nucleoside 5' triphosphate. The guanylyl cyclases, the mammalian adenylyl cyclases, and their microbial homologues function as pairs of homologous catalytic domains. The crystal structure of the rat type II adenylyl cyclase C2 catalytic domain was used to model by homology a mammalian adenylyl cyclase C1-C2 domain pair, a homodimeric adenylyl cyclase of Dictyostelium discoideum, a heterodimeric soluble guanylyl cyclase, and a homodimeric membrane guanylyl cyclase. Mg2+ATP or Mg2+GTP were docked into the active sites based on known stereochemical constraints on their conformation. The models are consistent with the activities of seven active site mutants. Asp-310 and Glu-432 of type I adenylyl cyclase coordinate a Mg2+ ion. The D310S and D310A mutants have 10-fold reduced Vmax and altered [Mg2+] dependence. The NTP purine moieties bind in mostly hydrophobic pockets. Specificity is conferred by a Lys and an Asp in adenylyl cyclase, and a Glu, an Arg, and a Cys in guanylyl cyclase. The models predict that an Asp from one domain is a general base in the reaction, and that the transition state is stabilized by a conserved Asn-Arg pair on the other domain. PMID- 9391040 TI - Identification of a second aryl phosphate-binding site in protein-tyrosine phosphatase 1B: a paradigm for inhibitor design. AB - The structure of the catalytically inactive mutant (C215S) of the human protein tyrosine phosphatase 1B (PTP1B) has been solved to high resolution in two complexes. In the first, crystals were grown in the presence of bis-(para phosphophenyl) methane (BPPM), a synthetic high-affinity low-molecular weight nonpeptidic substrate (Km = 16 microM), and the structure was refined to an R factor of 18. 2% at 1.9 A resolution. In the second, crystals were grown in a saturating concentration of phosphotyrosine (pTyr), and the structure was refined to an R-factor of 18.1% at 1.85 A. Difference Fourier maps showed that BPPM binds PTP1B in two mutually exclusive modes, one in which it occupies the canonical pTyr-binding site (the active site), and another in which a phosphophenyl moiety interacts with a set of residues not previously observed to bind aryl phosphates. The identification of a second pTyr molecule at the same site in the PTP1B/C215S pTyr complex confirms that these residues constitute a low-affinity noncatalytic aryl phosphate-binding site. Identification of a second aryl phosphate binding site adjacent to the active site provides a paradigm for the design of tight binding, highly specific PTP1B inhibitors that can span both the active site and the adjacent noncatalytic site. This design can be achieved by tethering together two small ligands that are individually targeted to the active site and the proximal noncatalytic site. PMID- 9391041 TI - Inhibition of HIV type 1 infectivity by constrained alpha-helical peptides: implications for the viral fusion mechanism. AB - Linear peptides derived from the membrane proximal region of the gp41 ectodomain are effective inhibitors of HIV type 1 (HIV-1)-mediated fusion events. These inhibitory peptides lack structure in solution, rendering mechanistic interpretation of their activity difficult. Using structurally constrained analogs of these molecules, we demonstrate that the peptides inhibit infectivity by adopting a helical conformation. Moreover, we show that a specific face of the helix must be exposed to block viral infectivity. Recent crystal structures show that the region of gp41 corresponding to the inhibitory peptides is helical and uses the analogous face to pack against a groove formed by an N-terminal coiled coil trimer. Our results provide a direct link between the inhibition of HIV-1 infectivity by these peptides and the x-ray structures, and suggest that the conformation of gp41 observed by crystallography represents the fusogenic state. Other agents that block HIV-1 infectivity by binding to this groove may hold promise for the treatment of AIDS. PMID- 9391042 TI - Overexpression of a glutamate receptor (GluR2) ligand binding domain in Escherichia coli: application of a novel protein folding screen. AB - Expression of the S1S2 ligand binding domain [Kuusinen, A., Arvola, M. & Keinanen, K. (1995) EMBO J. 14, 6327-6332] of the rat alpha-amino-3-hydroxy-5 methylisoxazole-4-propionic acid-selective glutamate receptor GluR2 in Escherichia coli under control of a T7 promoter leads to production of >100 mg/liter of histidine-tagged S1S2 protein (HS1S2) in the form of inclusion bodies. Using a novel fractional factorial folding screen and a rational, step-by step approach, multiple conditions were determined for the folding of the HS1S2 alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid binding domain. Characterization of the HS1S2 ligand binding domain showed that it is water soluble, monomeric, has significant secondary structure, and is sensitive to trypsinolysis at sites close to the beginning of the putative transmembrane regions. Application of a fractional factorial folding screen to other proteins may provide a useful means to evaluate E. coli as an economical and convenient expression host. PMID- 9391043 TI - Regulation of ribonuclease III processing by double-helical sequence antideterminants. AB - The double helix is a ubiquitous feature of RNA molecules and provides a target for nucleases involved in RNA maturation and decay. Escherichia coli ribonuclease III participates in maturation and decay pathways by site-specifically cleaving double-helical structures in cellular and viral RNAs. The site of cleavage can determine RNA functional activity and half-life and is specified in part by local tertiary structure elements such as internal loops. The involvement of base pair sequence in determining cleavage sites is unclear, because RNase III can efficiently degrade polymeric double-stranded RNAs of low sequence complexity. An alignment of RNase III substrates revealed an exclusion of specific Watson-Crick bp sequences at defined positions relative to the cleavage site. Inclusion of these "disfavored" sequences in a model substrate strongly inhibited cleavage in vitro by interfering with RNase III binding. Substrate cleavage also was inhibited by a 3-bp sequence from the selenocysteine-accepting tRNASec, which acts as an antideterminant of EF-Tu binding to tRNASec. The inhibitory bp sequences, together with local tertiary structure, can confer site specificity to cleavage of cellular and viral substrates without constraining the degradative action of RNase III on polymeric double-stranded RNA. Base pair antideterminants also may protect double-helical elements in other RNA molecules with essential functions. PMID- 9391044 TI - The C9 methyl group of retinal interacts with glycine-121 in rhodopsin. AB - The visual pigment rhodopsin is a prototypical G protein-coupled receptor. These receptors have seven transmembrane helices and are activated by specific receptor ligand interactions. Rhodopsin is unusual in that its retinal prosthetic group serves as an antagonist in the dark in the 11-cis conformation but is rapidly converted to an agonist on photochemical cis to trans isomerization. Receptor ligand interactions in rhodopsin were studied in the light and dark by regenerating site-directed opsin mutants with synthetic retinal analogues. A progressive decrease in light-dependent transducin activity was observed when a mutant opsin with a replacement of Gly121 was regenerated with 11-cis-retinal analogues bearing progressively larger R groups (methyl, ethyl, propyl) at the C9 position of the polyene chain. A progressive decrease in light activity was also observed as a function of increasing size of the residue at position 121 for both the 11-cis-9-ethyl- and the 11-cis-9-propylretinal pigments. In contrast, a striking increase of receptor activity in the dark-i.e., without chromophore isomerization-was observed when the molecular volume at either position 121 of opsin or C9 of retinal was increased. The ability of bulky replacements at either position to hinder ligand incorporation and to activate rhodopsin in the dark suggests a direct interaction between these two sites. A molecular model of the retinal-binding site of rhodopsin is proposed that illustrates the specific interaction between Gly121 and the C9 methyl group of 11-cis-retinal. Steric interactions in this region of rhodopsin are consistent with the proposal that movement of transmembrane helices 3 and 6 is concomitant with receptor activation. PMID- 9391045 TI - The major chromatin protein histone H1 binds preferentially to cis-platinum damaged DNA. AB - Both cis-diamminedichloroplatinum(II) (cisplatin or cis-DDP) and trans diamminedichloroplatinum(II) form covalent adducts with DNA. However, only the cis isomer is a potent anticancer agent. It has been postulated that the selective action of cis-DDP occurs through specific binding of nuclear proteins to cis-DDP-damaged DNA sites and that binding blocks DNA repair. We find that a very abundant nuclear protein, the linker histone H1, binds much more strongly to cis-platinated DNA than to trans-platinated or unmodified DNA. In competition experiments, H1 is shown to bind much more strongly than HMG1, which had been previously considered a major candidate for such binding in vivo. PMID- 9391046 TI - Structure of the recombinant full-length hamster prion protein PrP(29-231): the N terminus is highly flexible. AB - The prion diseases seem to be caused by a conformational change of the prion protein (PrP) from the benign cellular form PrPC to the infectious scrapie form PrPSc; thus, detailed information about PrP structure may provide essential insights into the mechanism by which these diseases develop. In this study, the secondary structure of the recombinant Syrian hamster PrP of residues 29-231 [PrP(29-231)] is investigated by multidimensional heteronuclear NMR. Chemical shift index analysis and nuclear Overhauser effect data show that PrP(29-231) contains three helices and possibly one short beta-strand. Most striking is the random-coil nature of chemical shifts for residues 30-124 in the full-length PrP. Although the secondary structure elements are similar to those found in mouse PrP fragment PrP(121-231), the secondary structure boundaries of PrP(29-231) are different from those in mouse PrP(121-231) but similar to those found in the structure of Syrian hamster PrP(90-231). Comparison of resonance assignments of PrP(29-231) and PrP(90-231) indicates that there may be transient interactions between the additional residues and the structured core. Backbone dynamics studies done by using the heteronuclear [1H]-15N nuclear Overhauser effect indicate that almost half of PrP(29-231), residues 29-124, is highly flexible. This plastic region could feature in the conversion of PrPC to PrPSc by template assisted formation of beta-structure. PMID- 9391047 TI - Histones H3 and H4 are components of upstream activation factor required for the high-level transcription of yeast rDNA by RNA polymerase I. AB - RNA polymerase I (Pol I) transcription in the yeast Saccharomyces cerevisiae is greatly stimulated in vivo and in vitro by the multiprotein complex, upstream activation factor (UAF). UAF binds tightly to the upstream element of the rDNA promoter, such that once bound (in vitro), UAF does not readily exchange onto a competing template. Of the polypeptides previously identified in purified UAF, three are encoded by genes required for Pol I transcription in vivo: RRN5, RRN9, and RRN10. Two others, p30 and p18, have remained uncharacterized. We report here that the N-terminal amino acid sequence, its mobility in gel electrophoresis, and the immunoreactivity of p18 shows that it is histone H3. In addition, histone H4 was found in UAF, and myc-tagged histone H4 could be used to affinity-purify UAF. Histones H2A and H2B were not detectable in UAF. These results suggest that histones H3 and H4 probably account for the strong binding of UAF to DNA and may offer a means by which general nuclear regulatory signals could be transmitted to Pol I. PMID- 9391048 TI - Metabolism of an insect diuretic hormone by Malpighian tubules studied by liquid chromatography coupled with electrospray ionization mass spectrometry. AB - The larger of two diuretic hormones of the tobacco hornworm, Manduca sexta, (Mas DH) is a peptide of 41 residues. It is one of a family of seven currently known insect diuretic hormones that are similar to the corticotropin-releasing factor urotensin-sauvagine family of peptides. We investigated the possible inactivation of Mas-DH by incubating it in vitro with larval Malpighian tubules (Mt), the target organ of the hormone. The medium was analyzed, and degradation products were identified, using on-line microbore reversed-phase liquid chromatography coupled to electrospray ionization mass spectrometry (RPLC-ESI-MS). This sensitive technique allows identification of metabolites of Mas-DH (present at an initial level of approximately 1 microM). An accurate Mr value for a metabolite is usually sufficient for unambiguous identification. Mas-DH is cleaved by Mt proteases initially at L29-R30 and R30-A31 under our assay conditions; some Mas DH is also oxidized, apparently at M2 and M11. The proteolysis can be inhibited by 5 mM EDTA, suggesting that divalent metals are needed for peptide cleavage. The oxidation of the hormone can be inhibited by catalase or 1 mM methionine, indicating that H2O2 or related reactive oxygen species are responsible for the oxidative degradation observed. RPLC-ESI-MS is shown here to be an elegant and efficient method for studying peptide hormone metabolism resulting from unknown proteases and pathways. PMID- 9391049 TI - Studies on the enzymatic and transcriptional activity of the dimerization cofactor for hepatocyte nuclear factor 1. AB - The relationship between the enzymatic and the transcriptional activity of the bifunctional protein pterin-4a-carbinolamine dehydratase/dimerization cofactor for hepatocyte nuclear factor 1 (DCoH) has been elucidated by site-directed mutagenesis. DCoH dimers harbor a binding site for hepatocyte nuclear factor 1 (HNF1), two active centers that bind pterins, and a saddle-shaped surface that resembles nucleic acid binding domains. Two domains of the protein have been selectively targeted to determine if a change in one activity affects the other. No strong correlation has been found, supporting the idea that carbinolamine dehydratase activity is not required for HNF1 binding in vitro or transcriptional coactivation in vivo. Double mutations in the active center, however, influence the in vivo transcriptional activity but not HNF1 binding. This finding suggests that some active center residues also are used during transcription, possibly for binding of another (macro)molecule. Several mutations in the saddle led to a surprising increase in transcription, therefore linking this domain to transcriptional regulation as well. The transcriptional function of DCoH therefore is composed of two parts, HNF1 binding and another contributing effect that involves the active site and, indirectly, the saddle. PMID- 9391051 TI - A mutant RNA polymerase that forms unusual open promoter complexes. AB - We describe a mutant Escherichia coli RNA polymerase (RNAP) that forms stable open promoter complexes even at -20 degrees C but with a shortened melted region that extends downstream to only position -7. In the presence of initiating transcription substrates, the mutant RNAP undergoes a temperature-dependent isomerization, resulting in a promoter complex that is indistinguishable from the wild-type RNAP-promoter complex, with the melted region extended downstream to position +4. We propose that the open complex formed by the mutant RNAP represents an intermediate on the normal promoter-opening pathway and that our results support earlier findings that initial promoter opening occurs in the upstream region of the -10 promoter consensus element and subsequently extends downstream to encompass the transcription start site. PMID- 9391050 TI - Bidirectional binding of the TATA box binding protein to the TATA box. AB - By selective attachment of a DNA cleavage agent to specific residues in the yeast TATA box binding protein (yTBP), we demonstrate that, in solution, yTBP binds to the TATA boxes of both the adenovirus major late promoter and the yeast CYC1 promoter with only a modest preference in orientation and binds well to several overlapping binding sites. The general factors TFIIA and TFIIB each increase the rotational and translational selectivity of yTBP but are not sufficient, at least individually, to confer a unique polarity to the preinitiation complex. We conclude that TBP alone cannot define the productive orientation of general factor assembly on a promoter. PMID- 9391052 TI - B12-dependent ribonucleotide reductases from deeply rooted eubacteria are structurally related to the aerobic enzyme from Escherichia coli. AB - The ribonucleotide reductases from three ancient eubacteria, the hyperthermophilic Thermotoga maritima (TM), the radioresistant Deinococcus radiodurans (DR), and the thermophilic photosynthetic Chloroflexus aurantiacus, were found to be coenzyme-B12 (class II) enzymes, similar to the earlier described reductases from the archaebacteria Thermoplasma acidophila and Pyrococcus furiosus. Reduction of CDP by the purified TM and DR enzymes requires adenosylcobalamin and DTT. dATP is a positive allosteric effector, but stimulation of the TM enzyme only occurs close to the temperature optimum of 80 90 degrees C. The TM and DR genes were cloned by PCR from peptide sequence information. The TM gene was sequenced completely and expressed in Escherichia coli. The deduced amino acid sequences of the two eubacterial enzymes are homologous to those of the archaebacteria. They can also be aligned to the sequence of the large protein of the aerobic E. coli ribonucleotide reductase that belongs to a different class (class I), which is not dependent on B12. Structure determinations of the E. coli reductase complexed with substrate and allosteric effectors earlier demonstrated a 10-stranded beta/alpha-barrel in the active site. From the conservation of substrate- and effector-binding residues we propose that the B12-dependent class II enzymes contain a similar barrel. PMID- 9391053 TI - Computer-based analysis of the binding steps in protein complex formation. AB - Computer models were used to examine whether and under what conditions the multimeric protein complex is inhibited by high concentrations of one of its components-an effect analogous to the prozone phenomenon in precipitin tests. A series of idealized simple "ball-and-stick" structures representing small oligomeric complexes of protein molecules formed by reversible binding reactions were analyzed to determine the binding steps leading to each structure. The equilibrium state of each system was then determined over a range of starting concentrations and Kds and the steady-state concentration of structurally complete oligomer calculated for each situation. A strong inhibitory effect at high concentrations was shown by any protein molecule forming a bridge between two or more separable parts of the complex. By contrast, proteins linked to the outside of the complex by a single bond showed no inhibition whatsoever at any concentration. Nonbridging, multivalent proteins in the body of the complex could show an inhibitory effect or not depending on the structure of the complex and the strength of its bonds. On the basis of this study, we suggest that the prozone phenomenon will occur widely in living cells and that it could be a crucial factor in the regulation of protein complex formation. PMID- 9391054 TI - Ultraspiracle: an invertebrate nuclear receptor for juvenile hormones. AB - Juvenile hormones (JH), a sesquiterpenoid group of ligands that regulate developmental transitions in insects, bind to the nuclear receptor ultraspiracle (USP). In fluorescence-based binding assays, USP protein binds JH III and JH III acid with specificity, adopting for each ligand a different final conformational state. JH III treatment of Saccharomyces cerevisiae expressing a LexA-USP fusion protein stabilizes an oligomeric association containing this protein, as detected by formation of a protein-DNA complex, and induces USP-dependent transcription in a reporter assay. We propose that regulation of morphogenetic transitions in invertebrates involves binding of JH or JH-like structures to USP. PMID- 9391056 TI - Coagulation initiated on herpesviruses. AB - Herpesviruses have been previously correlated to vascular disease and shown to cause thrombogenic and atherogenic changes to host cells. Herein we show that even in the absence of cells, purified cytomegalovirus (CMV) and herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) can initiate thrombin production. Functional assays demonstrated that purified HSV-1 and HSV-2 provide the necessary phospholipid (proPL) for assembling the coagulation factors Xa and Va into prothrombinase, which is responsible for generating thrombin. These observations are consistent with our earlier studies involving CMV. The presence of proPL on all three herpesviruses was confirmed directly by flow cytometry and electron microscopy by using annexin V and factor Va, respectively, as proPL specific probes. Of equal importance, we found that CMV, HSV-1, and HSV-2 were also able to facilitate factor Xa generation from the inactive precursor factor X, but only when factor VII/VIIa and Ca2+ were present. Monoclonal antibodies specific for tissue factor (TF), the coagulation initiator, inhibited this factor X activation and, furthermore, enabled identification of TF antigen on each virus type by flow cytometry and electron microscopy. Collectively, these data show that CMV, HSV-1, and HSV-2 can initiate the generation of thrombin by having essential proPL and TF activities on their surface. Unlike the normal cellular source, the viral activity is constitutive and, therefore, not restricted to sites of vascular injury. Thus cell-independent thrombin production may be the earliest event in vascular pathology mediated by herpesviruses. PMID- 9391055 TI - Glucocorticoid-mediated repression of nuclear factor-kappaB-dependent transcription involves direct interference with transactivation. AB - Glucocorticoids exert multiple anti-inflammatory activities, one of which is the inhibition of transcription dependent on the nuclear factor (NF)-kappaB. It has been suggested that the effect of dexamethasone (DEX), a glucocorticoid analog, is attributed to an increased production of the inhibitory IkappaB molecule, which in turn would bind and remove activated, DNA-bound NF-kappaB complexes in the cell nucleus. Upon investigating DEX-mediated repression of interleukin-6 expression induced by tumor necrosis factor, DEX treatment was found to act directly on NF-kappaB-dependent transcription, without changing the expression level of IkappaB. Neither the mRNA of IkappaB nor the protein was significantly elevated by a combined treatment with tumor necrosis factor and DEX of murine endothelial or fibroblast cells. The DNA-binding activity of induced NF-kappaB also remained unchanged after stimulation of cells with DEX. Evidence for a direct nuclear mechanism of action was obtained by analysis of cell lines stably expressing a fusion protein between the DNA-binding domain of the yeast Gal4 protein and the transactivating p65 subunit of NF-kappaB. Expression of a Gal4 dependent luciferase reporter gene activated by this nuclear fusion protein was also strongly repressed after addition of DEX. Because the DNA-binding activity of the Gal4 fusion protein was not affected by DEX, it can be concluded that the reduction of gene activation was caused by interference of the activated glucocorticoid receptor with the transactivation potential of the NF-kappaB p65 subunit. PMID- 9391057 TI - Topology of allosteric regulation of lactose permease. AB - Sugar transport by some permeases in Escherichia coli is allosterically regulated by the phosphorylation state of the intracellular regulatory protein, enzyme IIAglc of the phosphoenolpyruvate:sugar phosphotransferase system. A sensitive radiochemical assay for the interaction of enzyme IIAglc with membrane-associated lactose permease was used to characterize the binding reaction. The binding is stimulated by transportable substrates such as lactose, melibiose, and raffinose, but not by sugars that are not transported (maltose and sucrose). Treatment of lactose permease with N-ethylmaleimide, which blocks ligand binding and transport by alkylating Cys-148, also blocks enzyme IIAglc binding. Preincubation with the substrate analog beta-D-galactopyranosyl 1-thio-beta-D-galactopyranoside protects both lactose transport and enzyme IIAglc binding against inhibition by N ethylmaleimide. A collection of lactose permease replacement mutants at Cys-148 showed, with the exception of C148V, a good correlation of relative transport activity and enzyme IIAglc binding. The nature of the interaction of enzyme IIAglc with the cytoplasmic face of lactose permease was explored. The N- and C termini, as well as five hydrophilic loops in the permease, are exposed on the cytoplasmic surface of the membrane and it has been proposed that the central cytoplasmic loop of lactose permease is the major determinant for interaction with enzyme IIAglc. Lactose permease mutants with polyhistidine insertions in cytoplasmic loops IV/V and VI/VII and periplasmic loop VII/VIII retain transport activity and therefore substrate binding, but do not bind enzyme IIAglc, indicating that these regions of lactose permease may be involved in recognition of enzyme IIAglc. Taken together, these results suggest that interaction of lactose permease with substrate promotes a conformational change that brings several cytoplasmic loops into an arrangement optimal for interaction with the regulatory protein, enzyme IIAglc. A topological map of the proposed interaction is presented. PMID- 9391058 TI - Correct protein folding in glycerol. AB - Water is the natural medium for protein folding, which is also used in all in vitro studies. In the present work, we posed, and answered affirmatively, a question of whether it is possible to fold correctly a typical protein in a nonaqueous solvent. To this end, unfolded and reduced hen egg-white lysozyme was refolded and reoxidized in glycerol containing varying amounts of water. The unfolded/reduced enzyme was found to regain spontaneously substantial catalytic activity even in the nearly anhydrous solvent; for example, the refolding yield in 99% glycerol was still some one-third of that in pure water, and one-half of that was regained even in 99.8% glycerol. The less than full recovery of the enzymatic activity in glycerol is, as in water, because of competing protein aggregation during the refolding. Lysozyme reoxidation in glycerol was successfully mediated by two dissimilar oxidizing systems, and the refolding yield was markedly affected by the pH of the last aqueous solution before the transfer into glycerol. No recovery of the lysozyme activity was observed when the refolding/reoxidation reaction was carried out in the denaturing solvent dimethyl sulfoxide. This study paves the way for a systematic investigation of the solvent effect on protein folding and demonstrates that water is not a unique milieu for this process. PMID- 9391059 TI - Suppressor mutations in Escherichia coli methionyl-tRNA formyltransferase: role of a 16-amino acid insertion module in initiator tRNA recognition. AB - The specific formylation of initiator methionyl-tRNA by methionyl-tRNA formyltransferase (MTF; EC 2.1.2.9) is important for the initiation of protein synthesis in eubacteria and in eukaryotic organelles. The determinants for formylation in the tRNA are clustered mostly in the acceptor stem. As part of studies on the molecular mechanism of recognition of the initiator tRNA by MTF, we report here on the isolation and characterization of suppressor mutations in Escherichia coli MTF, which compensate for the formylation defect of a mutant initiator tRNA, lacking a critical determinant in the acceptor stem. We show that the suppressor mutant in MTF has a glycine-41 to arginine change within a 16 amino acid insertion found in MTF from many sources. A mutant with glycine-41 changed to lysine also acts as a suppressor, whereas mutants with changes to aspartic acid, glutamine, and leucine do not. The kinetic parameters of the purified wild-type and mutant Arg-41 and Lys-41 enzymes, determined by using the wild-type and mutant tRNAs as substrates, show that the Arg-41 and Lys-41 mutant enzymes compensate specifically for the strong negative effect of the acceptor stem mutation on formylation. These and other considerations suggest that the 16 amino acid insertion in MTF plays an important role in the specific recognition of the determinants for formylation in the acceptor stem of the initiator tRNA. PMID- 9391060 TI - The nucleocapsid protein specifically anneals tRNALys-3 onto a noncomplementary primer binding site within the HIV-1 RNA genome in vitro. AB - HIV type 1 (HIV-1) specifically uses host cell tRNALys-3 as a primer for reverse transcription. The 3' 18 nucleotides of this tRNA are complementary to a region on the HIV RNA genome known as the primer binding site (PBS). HIV-1 has a strong preference for maintaining a lysine-specific PBS in vivo, and viral genomes with mutated PBS sequences quickly revert to be complementary to tRNALys-3. To investigate the mechanism for the observed PBS reversion events in vitro, we examined the capability of the nucleocapsid protein (NC) to anneal various tRNA primer sequences onto either complementary or noncomplementary PBSs. We show that NC can anneal different full-length tRNAs onto viral RNA transcripts derived from the HIV-1 MAL or HXB2 isolates, provided that the PBS is complementary to the tRNA used. In contrast, NC promotes specific annealing of only tRNALys-3 onto an RNA template (HXB2) whose PBS sequence has been mutated to be complementary to the 3' 18 nt of human tRNAPro. Moreover, HIV-1 reverse transcriptase extends this binary complex from the proline-specific PBS. The formation of the noncomplementary binary complex does not occur when a chimeric tRNALys/Pro containing proline-specific D and anticodon domains is used as the primer. Thus, elements outside the acceptor-TPsiC domains of tRNALys-3 play an important role in preferential primer use in vitro. Our results support the hypothesis that mutant PBS reversion is a result of tRNALys-3 annealing onto and extension from a PBS that specifies an alternate host cell tRNA. PMID- 9391061 TI - Functional separation of pre-rRNA processing steps revealed by truncation of the U3 small nucleolar ribonucleoprotein component, Mpp10. AB - The U3 small nucleolar ribonucleoprotein (snoRNP) is required for three cleavage events that generate the mature 18S rRNA from the pre-rRNA. In Saccharomyces cerevisiae, depletion of Mpp10, a U3 snoRNP-specific protein, halts 18S rRNA production and impairs cleavage at the three U3 snoRNP-dependent sites: A0, A1, and A2. We have identified truncation mutations of Mpp10 that affect 18S rRNA synthesis and confer cold-sensitivity and slow growth. However, distinct from yeast cells depleted of Mpp10, the mutants carrying these truncated Mpp10 proteins accumulate a novel precursor, resulting from cleavage at only A0. The Mpp10 truncations do not alter association of Mpp10 with the U3 snoRNA, nor do they affect snoRNA or protein stability. Thus, the role in processing of the U3 snoRNP can be separated into cleavage at the A0 site, which occurs in the presence of truncated Mpp10, and cleavage at the A1/A2 sites, which occurs only with intact Mpp10. These results strongly argue for a role for Mpp10 in processing at the A1/A2 sites. PMID- 9391062 TI - Formation of stable adducts and absence of depurinating DNA adducts in cells and DNA treated with the potent carcinogen dibenzo[a,l]pyrene or its diol epoxides. AB - Polycyclic aromatic hydrocarbons (PAH) are widespread environmental contaminants, and some are potent carcinogens in rodents. Carcinogenic PAH are activated in cells to metabolites that react with DNA to form stable covalent DNA adducts. It has been proposed [Cavalieri, E. L. & Roger, E. G. (1995) Xenobiotica 25, 677 688] that unstable DNA adducts are also formed and that apurinic sites in the DNA resulting from unstable PAH adducts play a key role in the initiation of cancer. The potent carcinogen dibenzo[a,l]pyrene (DB[a, l]P) is activated in cells to (+) syn- and (-)-anti-DB[a,l]P-11, 12-diol-13,14-epoxide (DB[a,l]PDE), which have been shown to form stable adducts with DNA. To evaluate the importance of unstable PAH adducts, we compared stable adduct formation to apurinic site formation. Stable DB[a,l]PDE adducts were determined by 33P-postlabeling and HPLC. To measure apurinic sites they were converted to strand breaks, and these were monitored by examining the integrity of a particular restriction fragment of the dihydrofolate reductase gene. The method easily detected apurinic sites resulting from methylation by treatment of cells or DNA with dimethyl sulfate or from reaction of DNA with DB[a,l]P in the presence of horseradish peroxidase. We estimate the method could detect 0.1 apurinic site in the 14-kb fragment examined. However, apurinic sites were below our limit of detection in DNA treated directly with (+)-syn- or (-)-anti-DB[a,l]PDE or in DNA from Chinese hamster ovary B11 cells so treated, although in these samples the frequency of stable adducts ranged from 3 to 10 per 14 kb. We also treated the human mammary carcinoma cell line MCF-7 with DB[a,l]P and again could not detect significant amounts of unstable adducts. These results indicate that the proportion of stable adducts formed by DB[a,l]P activated in cells and its diol epoxides is greater than 99% and suggest a predominant role for stable DNA adducts in the carcinogenic activity of DB[a,l]P. PMID- 9391063 TI - Escherichia coli RNA polymerase terminates transcription efficiently at rho independent terminators on single-stranded DNA templates. AB - Several models have been proposed for the mechanism of transcript termination by Escherichia coli RNA polymerase at rho-independent terminators. Yager and von Hippel (Yager, T. D. & von Hippel, P. H. (1991) Biochemistry 30, 1097-118) postulated that the transcription complex is stabilized by enzyme-nucleic acid interactions and the favorable free energy of a 12-bp RNA-DNA hybrid but is destabilized by the free energy required to maintain an extended transcription bubble. Termination, by their model, is viewed simply as displacement of the RNA transcript from the hybrid helix by reformation of the DNA helix. We have proposed an alternative model where the RNA transcript is stably bound to RNA polymerase primarily through interactions with two single-strand specific RNA binding sites; termination is triggered by formation of an RNA hairpin that reduces binding of the RNA to one RNA-binding site and, ultimately, leads to its ejection from the complex. To distinguish between these models, we have tested whether E. coli RNA polymerase can terminate transcription at rho-independent terminators on single-stranded DNA. RNA polymerase cannot form a transcription bubble on these templates; thus, the Yager-von Hippel model predicts that intrinsic termination will not occur. We find that transcript elongation on single-stranded DNA templates is hindered somewhat by DNA secondary structure. However, E. coli RNA polymerase efficiently terminates and releases transcripts at several rho-independent terminators on such templates at the same positions as termination occurs on duplex DNAs. Therefore, neither the nontranscribed DNA strand nor the transcription bubble is essential for rho-independent termination by E. coli RNA polymerase. PMID- 9391064 TI - A direct electrode-driven P450 cycle for biocatalysis. AB - The large potential of redox enzymes to carry out formation of high value organic compounds motivates the search for innovative strategies to regenerate the cofactors needed by their biocatalytic cycles. Here, we describe a bioreactor where the reducing power to the cycle is supplied directly to purified cytochrome CYP101 (P450cam; EC 1.14.15.1) through its natural redox partner (putidaredoxin) using an antimony-doped tin oxide working electrode. Required oxygen was produced at a Pt counter electrode by water electrolysis. A continuous catalytic cycle was sustained for more than 5 h and 2,600 enzyme turnovers. The maximum product formation rate was 36 nmol of 5-exo-hydroxycamphor/nmol of CYP101 per min. PMID- 9391065 TI - Synthesis and characterization of a novel retinylamine analog inhibitor of constitutively active rhodopsin mutants found in patients with autosomal dominant retinitis pigmentosa. AB - Two different mutations of the active-site Lys-296 in rhodopsin, K296E and K296M, have been found to cause autosomal dominant retinitis pigmentosa (ADRP). In vitro studies have shown that both mutations result in constitutive activation of the protein, suggesting that the activated state of the receptor may be responsible for retinal degeneration in patients with these mutations. Previous work has highlighted the potential of retinylamine analogs as active-site directed inactivators of constitutively active mutants of rhodopsin with the idea that these or related compounds might be used therapeutically for cases of ADRP involving mutations of the active-site Lys. Unfortunately, however, amine derivatives of 11-cis-retinal, although highly effective against a K296G mutant of rhodopsin, were without affect on the two naturally occurring ADRP mutants, presumably because of the greater steric bulk of Glu and Met side chains in comparison to Gly. For this reason we synthesized a retinylamine analog one carbon shorter than the parent 11-cis-retinal and show that this compound is indeed an effective inhibitor of both the K296E and K296M mutants. The 11-cis C19 retinylamine analog 1 inhibits constitutive activation of transducin by these mutants and their constitutive phosphorylation by rhodopsin kinase, and it does so in the presence of continuous illumination from room lights. PMID- 9391066 TI - A terbenzimidazole that preferentially binds and conformationally alters structurally distinct DNA duplex domains: a potential mechanism for topoisomerase I poisoning. AB - The terbenzimidazoles are a class of synthetic ligands that poison the human topoisomerase I (TOP1) enzyme and promote cancer cell death. It has been proposed that drugs of this class act as TOP1 poisons by binding to the minor groove of the DNA substrate of TOP1 and altering its structure in a manner that results in enzyme-mediated DNA cleavage. To test this hypothesis, we characterize and compare the binding properties of a 5-phenylterbenzimidazole derivative (5PTB) to the d(GA4T4C)2 and d(GT4A4C)2 duplexes. The d(GA4T4C)2 duplex contains an uninterrupted 8-bp A.T domain, which, on the basis of x-ray crystallographic data, should induce a highly hydrated "A-tract" conformation. This duplex also exhibits anomalously slow migration in a polyacrylamide gel, a feature characteristic of a noncanonical global conformational state frequently described as "bent." By contrast, the d(GT4A4C)2 duplex contains two 4-bp A.T tracts separated by a TpA dinucleotide step, which should induce a less hydrated "B like" conformation. This duplex also migrates normally in a polyacrylamide gel, a feature further characteristic of a global, canonical B-form duplex. Our data reveal that, at 20 degrees C, 5PTB exhibits an approximately 2. 3 kcal/mol greater affinity for the d(GA4T4C)2 duplex than for the d(GT4A4C)2 duplex. Significantly, we find this sequence/conformational binding specificity of 5PTB to be entropic in origin, an observation consistent with a greater degree of drug binding-induced dehydration of the more solvated d(GA4T4C)2 duplex. By contrast with the differential duplex affinity exhibited by 5PTB, netropsin and 4',6 diamidino-2-phenylindole (DAPI), two AT-specific minor groove binding ligands that are inactive as human TOP1 poisons, bind to both duplexes with similar affinities. The electrophoretic behaviors of the ligand-free and ligand-bound duplexes are consistent with 5PTB-induced bending and/or unwinding of both duplexes, which, for the d(GA4T4C)2 duplex, is synergistic with the endogenous sequence-directed electrophoretic properties of the ligand-free duplex state. By contrast, the binding to either duplex of netropsin or DAPI induces little or no change in the electrophoretic mobilities of the duplexes. Our results demonstrate that the TOP1 poison 5PTB binds differentially to and alters the structures of the two duplexes, in contrast to netropsin and DAPI, which bind with similar affinities to the two duplexes and do not significantly alter their structures. These results are consistent with a mechanism for TOP1 poisoning in which drugs such as 5PTB differentially target conformationally distinct DNA sites and induce structural changes that promote enzyme-mediated DNA cleavage. PMID- 9391067 TI - Repression of transcription mediated by dual elements in the CCAAT/enhancer binding protein alpha gene. AB - During adipocyte differentiation, the expression of C/EBPalpha is activated, which in turn serves to transcriptionally activate numerous adipocyte genes. A previous search for cis elements that regulate transcription of the C/EBPalpha gene led to the identification of a potential repressive element within the proximal 5' flanking region of the gene. Nuclear extracts from 3T3-L1 preadipocytes, but not adipocytes, were found to contain a factor, CUP (C/EBPalpha undifferentiated protein), that binds to this site (the CUP-1 site). In the present investigation, we show that C/EBPalpha promoter-luciferase constructs containing both the proximal 5' flanking and the entire 5' untranslated regions of the gene exhibit an expression pattern during adipocyte differentiation comparable to that of the endogenous C/EBPalpha gene. Mutation of the CUP-1 site in these constructs had little effect on reporter gene expression; however, when this mutation was combined with deletion of the 5' untranslated region, reporter gene expression by preadipocytes was dramatically up-regulated. Consistent with this finding, a second CUP binding site (the CUP-2 site) was identified in the 5' untranslated region. Although mutation of either CUP element in constructs containing both the 5' flanking and 5' untranslated region had little effect on reporter gene transcription, mutation of both CUP elements markedly activated transcription. Thus, it appears that dual CUP regulatory elements repress transcription of the C/EBPalpha gene prior to induction of the adipocyte differentiation program. PMID- 9391068 TI - Mode matches and their locations in the hydrophobic free energy sequences of peptide ligands and their receptor eigenfunctions. AB - Patterns in sequences of amino acid hydrophobic free energies predict secondary structures in proteins. In protein folding, matches in hydrophobic free energy statistical wavelengths appear to contribute to selective aggregation of secondary structures in "hydrophobic zippers." In a similar setting, the use of Fourier analysis to characterize the dominant statistical wavelengths of peptide ligands' and receptor proteins' hydrophobic modes to predict such matches has been limited by the aliasing and end effects of short peptide lengths, as well as the broad-band, mode multiplicity of many of their frequency (power) spectra. In addition, the sequence locations of the matching modes are lost in this transformation. We make new use of three techniques to address these difficulties: (i) eigenfunction construction from the linear decomposition of the lagged covariance matrices of the ligands and receptors as hydrophobic free energy sequences; (ii) maximum entropy, complex poles power spectra, which select the dominant modes of the hydrophobic free energy sequences or their eigenfunctions; and (iii) discrete, best bases, trigonometric wavelet transformations, which confirm the dominant spectral frequencies of the eigenfunctions and locate them as (absolute valued) moduli in the peptide or receptor sequence. The leading eigenfunction of the covariance matrix of a transmembrane receptor sequence locates the same transmembrane segments seen in n block-averaged hydropathy plots while leaving the remaining hydrophobic modes unsmoothed and available for further analyses as secondary eigenfunctions. In these receptor eigenfunctions, we find a set of statistical wavelength matches between peptide ligands and their G-protein and tyrosine kinase coupled receptors, ranging across examples from 13.10 amino acids in acid fibroblast growth factor to 2.18 residues in corticotropin releasing factor. We find that the wavelet-located receptor modes in the extracellular loops are compatible with studies of receptor chimeric exchanges and point mutations. A nonbinding corticotropin-releasing factor receptor mutant is shown to have lost the signatory mode common to the normal receptor and its ligand. Hydrophobic free energy eigenfunctions and their transformations offer new quantitative physical homologies in database searches for peptide-receptor matches. PMID- 9391069 TI - Relationship between the oxidation potential and electron spin density of the primary electron donor in reaction centers from Rhodobacter sphaeroides. AB - The primary electron donor in bacterial reaction centers is a dimer of bacteriochlorophyll a molecules, labeled L or M based on their proximity to the symmetry-related protein subunits. The electronic structure of the bacteriochlorophyll dimer was probed by introducing small systematic variations in the bacteriochlorophyll-protein interactions by a series of site-directed mutations that replaced residue Leu M160 with histidine, tyrosine, glutamic acid, glutamine, aspartic acid, asparagine, lysine, and serine. The midpoint potentials for oxidation of the dimer in the mutants showed an almost continuous increase up to approximately 60 mV compared with wild type. The spin density distribution of the unpaired electron in the cation radical state of the dimer was determined by electron-nuclear-nuclear triple resonance spectroscopy in solution. The ratio of the spin density on the L side of the dimer to the M side varied from approximately 2:1 to approximately 5:1 in the mutants compared with approximately 2:1 for wild type. The correlation between the midpoint potential and spin density distribution was described using a simple molecular orbital model, in which the major effect of the mutations is assumed to be a change in the energy of the M half of the dimer, providing estimates for the coupling and energy levels of the orbitals in the dimer. These results demonstrate that the midpoint potential can be fine-tuned by electrostatic interactions with amino acids near the dimer and show that the properties of the electronic structure of a donor or acceptor in a protein complex can be directly related to functional properties such as the oxidation-reduction midpoint potential. PMID- 9391070 TI - Hypersensitivity of Ku80-deficient cell lines and mice to DNA damage: the effects of ionizing radiation on growth, survival, and development. AB - We recently have shown that mice deficient for the 86-kDa component (Ku80) of the DNA-dependent protein kinase exhibit growth retardation and a profound deficiency in V(D)J (variable, diversity, and joining) recombination. These defects may be related to abnormalities in DNA metabolism that arise from the inability of Ku80 mutant cells to process DNA double-strand breaks. To further characterize the role of Ku80 in DNA double-strand break repair, we have generated embryonic stem cells and pre-B cells and examined their response to ionizing radiation. Ku80(-/ ) embryonic stem cells are more sensitive than controls to gamma-irradiation, and pre-B cells derived from Ku80 mutant mice display enhanced spontaneous and gamma ray-induced apoptosis. We then determined the effects of ionizing radiation on the survival, growth, and lymphocyte development in Ku80-deficient mice. Ku80(-/ ) mice display a hypersensitivity to gamma-irradiation, characterized by loss of hair pigmentation, severe injury to the gastrointestinal tract, and enhanced mortality. Exposure of newborn Ku80(-/-) mice to sublethal doses of ionizing radiation enhances their growth retardation and results in the induction of T cell-specific differentiation. However, unlike severe combined immunodeficient mice, radiation-induced T cell development in Ku80(-/-) mice is not accompanied by extensive thymocyte proliferation. The response of Ku80-deficient cell lines and mice to DNA-damaging agents provides important insights into the role of Ku80 in growth regulation, lymphocyte development, and DNA repair. PMID- 9391071 TI - Aldehyde dehydrogenase class 3 expression: identification of a cornea-preferred gene promoter in transgenic mice. AB - Aldehyde dehydrogenase class 3 (ALDH3) constitutes 20-40% of the total water soluble proteins in the mammalian cornea. Here, we show by Northern blot analysis that ALDH3 expression in the mouse is at least 500-fold higher in the cornea than in any other tissue examined, with very low levels of expression detected in the stomach, urinary bladder, ocular lens, and lung. Histochemical localization reveals that this exceptional level of expression in the mouse cornea occurs in the anterior epithelial cells and that little ALDH3 is present in the keratocytes or corneal endothelial cells. A 13-kbp mouse ALDH3 promoter fragment containing >12 kbp of the 5' flanking sequence, the 40-bp untranslated first exon, and 29 bp of intron 1 directed cat reporter gene expression to tissues that express the endogenous ALDH3 gene, except that transgene promoter activity was higher in the stomach and bladder than in the cornea. By contrast, when driven by a 4.4-kbp mouse ALDH3 promoter fragment [1,050-bp 5' flanking region, exon 1, intron 1 (3.4 kbp), and 7 bp of exon 2] expression of the cat reporter gene was confined to the corneal epithelial cells, except for very low levels in the liver, effectively reproducing the corneal expression pattern of the endogenous ALDH3 gene. These results indicate that tissue-specific expression of ALDH3 is determined by positive and negative elements in the 5' flanking region of the gene and suggests putative silencers located in intron 1. We demonstrate regulatory sequences capable of directing cornea-specific gene expression, affording the opportunity for genetic engineering in this transparent tissue. PMID- 9391072 TI - Scavenger receptor class B, type I (SR-BI) is the major route for the delivery of high density lipoprotein cholesterol to the steroidogenic pathway in cultured mouse adrenocortical cells. AB - The class B, type I scavenger receptor, SR-BI, binds high density lipoprotein (HDL) and mediates the selective uptake of HDL cholesteryl ester (CE) by cultured transfected cells. The high levels of SR-BI expression in steroidogenic cells in vivo and its regulation by tropic hormones provides support for the hypothesis that SR-BI is a physiologically relevant HDL receptor that supplies substrate cholesterol for steroid hormone synthesis. This hypothesis was tested by determining the ability of antibody directed against murine (m) SR-BI to inhibit the selective uptake of HDL CE in Y1-BS1 adrenocortical cells. Anti-mSR-BI IgG inhibited HDL CE-selective uptake by 70% and cell association of HDL particles by 50% in a dose-dependent manner. The secretion of [3H]steroids derived from HDL containing [3H]CE was inhibited by 78% by anti-mSR-BI IgG. These results establish mSR-BI as the major route for the selective uptake of HDL CE and the delivery of HDL cholesterol to the steroidogenic pathway in cultured mouse adrenal cells. PMID- 9391073 TI - Baculovirus inhibitors of apoptosis (IAPs) block activation of Sf-caspase-1. AB - We have investigated the ability of Sf-caspase-1 and two mammalian caspases, caspase-1 and caspase-3, to induce apoptosis in Spodoptera frugiperda Sf-21 insect cells. While the transient expression of the pro-Sf-caspase-1 did not induce apoptosis, expression of the pro-domain deleted form, p31, or coexpression of the two subunits of mature Sf-caspase-1, p19 and p12, induced apoptosis in Sf 21 cells. The behavior of Sf-caspase-1 resembled that of the closely related mammalian caspase, caspase-3, and contrasted with that of the mammalian caspase 1, the pro-form of which was active in inducing apoptosis in Sf-21 cells. The baculovirus caspase inhibitor P35 blocked apoptosis induced by active forms of all three caspases. In contrast, members of the baculovirus inhibitor of apoptosis (IAP) family failed to block active caspase-induced apoptosis. However, during viral infection, expression of OpIAP or CpIAP blocked the activation of pro-Sf-caspase-1 and the associated induction of apoptosis. Thus, the mechanism by which baculovirus IAPs inhibit apoptosis is distinct from the mechanism by which P35 blocks apoptosis and involves inhibition of the activation of pro caspases like Sf-caspase-1. PMID- 9391074 TI - Angiogenesis promoted by vascular endothelial growth factor: regulation through alpha1beta1 and alpha2beta1 integrins. AB - Vascular endothelial growth factor (VEGF), also known as vascular permeability factor, is a cytokine of central importance for the angiogenesis associated with cancers and other pathologies. Because angiogenesis often involves endothelial cell (EC) migration and proliferation within a collagen-rich extracellular matrix, we investigated the possibility that VEGF promotes neovascularization through regulation of collagen receptor expression. VEGF induced a 5- to 7-fold increase in dermal microvascular EC surface protein expression of two collagen receptors-the alpha1beta1 and alpha2beta1 integrins-through induction of mRNAs encoding the alpha1 and alpha2 subunits. In contrast, VEGF did not induce increased expression of the alpha3beta1 integrin, which also has been implicated in collagen binding. Integrin alpha1-blocking and alpha2-blocking antibodies (Ab) each partially inhibited attachment of microvascular EC to collagen I, and alpha1 blocking Ab also inhibited attachment to collagen IV and laminin-1. Induction of alpha1beta1 and alpha2beta1 expression by VEGF promoted cell spreading on collagen I gels which was abolished by a combination of alpha1-blocking and alpha2-blocking Abs. In vivo, a combination of alpha1-blocking and alpha2 blocking Abs markedly inhibited VEGF-driven angiogenesis; average cross-sectional area of individual new blood vessels was reduced 90% and average total new vascular area was reduced 82% without detectable effects on the pre-existing vasculature. These data indicate that induction of alpha1beta1 and alpha2beta1 expression by EC is an important mechanism by which VEGF promotes angiogenesis and that alpha1beta1 and alpha2beta1 antagonists may prove effective in inhibiting VEGF-driven angiogenesis in cancers and other important pathologies. PMID- 9391075 TI - Characterization and cell cycle regulation of the related human telomeric proteins Pin2 and TRF1 suggest a role in mitosis. AB - Telomeres are essential for preserving chromosome integrity during the cell cycle and have been specifically implicated in mitotic progression, but little is known about the signaling molecule(s) involved. The human telomeric repeat binding factor protein (TRF1) is shown to be important in regulating telomere length. However, nothing is known about its function and regulation during the cell cycle. The sequence of PIN2, one of three human genes (PIN1-3) we previously cloned whose products interact with the Aspergillus NIMA cell cycle regulatory protein kinase, reveals that it encodes a protein that is identical in sequence to TRF1 apart from an internal deletion of 20 amino acids; Pin2 and TRF1 may be derived from the same gene, PIN2/TRF1. However, in the cell Pin2 was found to be the major expressed product and to form homo- and heterodimers with TRF1; both dimers were localized at telomeres. Pin2 directly bound the human telomeric repeat DNA in vitro, and was localized to all telomeres uniformly in telomerase positive cells. In contrast, in several cell lines that contain barely detectable telomerase activity, Pin2 was highly concentrated at only a few telomeres. Interestingly, the protein level of Pin2 was highly regulated during the cell cycle, being strikingly increased in G2+M and decreased in G1 cells. Moreover, overexpression of Pin2 resulted in an accumulation of HeLa cells in G2+M. These results indicate that Pin2 is the major human telomeric protein and is highly regulated during the cell cycle, with a possible role in mitosis. The results also suggest that Pin2/TRF1 may connect mitotic control to the telomere regulatory machinery whose deregulation has been implicated in cancer and aging. PMID- 9391076 TI - The glyoxysomal and plastid molecular chaperones (70-kDa heat shock protein) of watermelon cotyledons are encoded by a single gene. AB - The monoclonal a-70-kDa heat shock protein (hsp70) antibody recognizes in crude extracts from watermelon (Citrullus vulgaris) cotyledons two hsps with molecular masses of 70 and 72 kDa. Immunocytochemistry on watermelon cotyledon tissue and on isolated glyoxysomes identified hsp70s in the matrix of glyoxysomes and plastids. Affinity purification and partial amino acid determination revealed the 70-kDa protein to share high sequence identity with cytosolic hsp70s from a number of plant species, while the 72 kDa protein was very similar to plastid hsp70s from pea and cucumber. A full-length cDNA clone encoding the 72-kDa hsp70 was isolated and identified two start methionines in frame within the N-terminal presequence leading either to an N-terminal extension of 67 amino acids or to a shorter one of 47 amino acids. The longer presequence was necessary and sufficient to target a reporter protein into watermelon proplastids in vitro. The shorter extension starting from the second methionine within the long version harbored a consensus peroxisomal targeting signal (RT-X5-KL) that directed in vivo a reporter protein into peroxisomes of the yeast Hansenula polymorpha. Peroxisomal targeting was however prevented, when the 67-residue presequence was fused to the reporter protein, indicating that the peroxisomal targeting signal 2 information is hidden in this context. We propose that the 72-kDa hsp70 is encoded by a single gene, but targeted alternatively into two organelles by the modulated use of its presequence. PMID- 9391077 TI - Opposing mitogenic and anti-mitogenic actions of parathyroid hormone-related protein in vascular smooth muscle cells: a critical role for nuclear targeting. AB - Parathyroid hormone-related protein (PTHrP) is a prohormone that is posttranslationally processed to a family of mature secretory forms, each of which has its own cognate receptor(s) on the cell surface that mediate the actions of PTHrP. In addition to being secreted via the classical secretory pathway and interacting with cell surface receptors in a paracrine/autocrine fashion, PTHrP appears to be able to enter the nucleus directly following translation and influence cellular events in an "intracrine" fashion. In this report, we demonstrate that PTHrP can be targeted to the nucleus in vascular smooth muscle cells, that this nuclear targeting is associated with a striking increase in mitogenesis, that this nuclear effect on proliferation is the diametric opposite of the effects of PTHrP resulting from interaction with cell surface receptors on vascular smooth muscle cells, and that the regions of the PTHrP sequence responsible for this nuclear targeting represent a classical bipartite nuclear localization signal. This report describes the activation of the cell cycle in association with nuclear localization of PTHrP in any cell type. These findings have important implications for the normal physiology of PTHrP in the many tissues which produce it, and suggest that gene delivery of PTHrP or modified variants may be useful in the management of atherosclerotic vascular disease. PMID- 9391078 TI - SARPs: a family of secreted apoptosis-related proteins. AB - Quiescent mouse embryonic C3H/10T1/2 cells are more resistant to different proapoptotic stimuli than are these cells in the exponential phase of growth. However, the exponentially growing 10T1/2 cells are resistant to inhibitors of RNA or protein synthesis, whereas quiescent cells die upon these treatments. Conditioned medium from quiescent 10T1/2 cells possesses anti-apoptotic activity, suggesting the presence of protein(s) that function as an inhibitor of the apoptotic program. Using differential display technique, we identified and cloned a cDNA designated sarp1 (secreted apoptosis-related protein) that is expressed in quiescent but not in exponentially growing 10T1/2 cells. Hybridization studies with sarp1 revealed two additional family members. Cloning and sequencing of sarp2 and sarp3 revealed 38% and 40% sequence identity to sarp1, respectively. Human breast adenocarcinoma MCF7 cells stably transfected with sarp1 or infected with SARP1-expressing adenovirus became more resistant, whereas cells transfected with sarp2 displayed increased sensitivity to different proapoptotic stimuli. Expression of sarp family members is tissue specific. sarp mRNAs encode secreted proteins that possess a cysteine-rich domain (CRD) homologous to the CRD of frizzled proteins but lack putative membrane-spanning segments. Expression of SARPs modifies the intracellular levels of beta-catenin, suggesting that SARPs interfere with the Wnt-frizzled proteins signaling pathway. PMID- 9391079 TI - Activation of hPAK65 by caspase cleavage induces some of the morphological and biochemical changes of apoptosis. AB - Apoptosis is a highly regulated form of cell death, characterized by distinctive features such as cellular shrinkage and nuclear condensation. We demonstrate here that proteolytic activation of hPAK65, a p21-activated kinase, induces morphological changes and elicits apoptosis. hPAK65 is cleaved both in vitro and in vivo by caspases at a single site between the N-terminal regulatory p21 binding domain and the C-terminal kinase domain. The C-terminal cleavage product becomes activated, with a kinetic profile that parallels caspase activation during apoptosis. This C-terminal hPAK65 fragment also activates the c-Jun N terminal kinase pathway in vivo. Microinjection or transfection of this truncated hPAK65 causes striking alterations in cellular and nuclear morphology, which subsequently promotes apoptosis in both CHO and Hela cells. Conversely, apoptosis is delayed in cells expressing a dominant-negative form of hPAK65. These findings provide a direct evidence that the activated form of hPAK65 generated by caspase cleavage is a proapoptotic effector that mediates morphological and biochemical changes seen in apoptosis. PMID- 9391080 TI - Expansion in vitro of adult murine hematopoietic stem cells with transplantable lympho-myeloid reconstituting ability. AB - Elucidation of mechanisms that regulate hematopoietic stem cell self-renewal and differentiation would be facilitated by the identification of defined culture conditions that allow these cells to be amplified. We now demonstrate a significant net increase (3-fold, P < 0.001) in vitro of cells that are individually able to permanently and competitively reconstitute the lymphoid and myeloid systems of syngeneic recipient mice when Sca-1(+)lin- adult marrow cells are incubated for 10 days in serum-free medium with interleukin 11, flt3-ligand, and Steel factor. Moreover, the culture-derived repopulating cells continued to expand their numbers in the primary hosts at the same rate seen in recipients of noncultured stem cells. In the expansion cultures, long-term culture-initiating cells increased 7- +/- 2-fold, myeloid colony-forming cells increased 140- +/- 36 fold, and total nucleated cells increased 230- +/- 62-fold. Twenty-seven of 100 cultures initiated with 15 Sca-1(+)lin- marrow cells were found to contain transplantable stem cells 10 days later. This frequency of positive cultures is the same as the frequency of transplantable stem cells in the original input suspension, suggesting that most had undergone at least one self-renewal division in vitro. No expansion of stem cells was seen when Sca-1+TER119- CD34+ day 14.5 fetal liver cells were cultured under the same conditions. These findings set the stage for further investigations of the mechanisms by which cytokine stimulation may elicit different outcomes in mitotically activated hematopoietic stem cells during ontogeny and in the adult. PMID- 9391081 TI - Structural features of the kringle domain determine the intracellular degradation of under-gamma-carboxylated prothrombin: studies of chimeric rat/human prothrombin. AB - Vitamin K antagonists such as warfarin inhibit the vitamin K-dependent gamma glutamyl carboxylation during protein processing and block the secretion of under gamma-carboxylated prothrombin (FII) in the rat but not in the human or bovine. Under-gamma-carboxylated prothrombin is also secreted from warfarin-treated human (HepG2) cell cultures but is degraded in the endoplasmic reticulum in warfarin treated rat (H-35) cell cultures. This differential response to warfarin has been shown to be determined by the structural difference in the proteins rather than by the origin of the cell line. When recombinant rat prothrombin (rFII) and human prothrombin (hFII) were expressed in a transformed human kidney cell line (HEK293), secretion of rFII but not hFII was drastically decreased in response to warfarin. To determine the structural signal required for this differential response, chimeric cDNAs with the propeptide/Gla domains, kringle domain, and serine protease domain exchanged between rFII and hFII were generated (FIIRHH and FIIHRR, FIIRRH and FIIHHR, FIIRHR and FIIHRH) and expressed in both warfarin treated HEK293 cells and HepG2 cells. The presence of the hFII kringle domain changed the stability of rFII to that of hFII, and the rFII kringle domain changed the stability of hFII to that of rFII. The kringle domain therefore is critical in determining the metabolic fate of under-gamma-carboxylated prothrombin precursors during processing. Prothrombin contains two kringle structures, and expression of additional rFII/hFII chimeras (FIIHrhH and FIIHhrH, FIIRrhR, and FIIRhrR) was used to determine that the first of the two kringles plays a more important role in the recognition process. PMID- 9391082 TI - Cell locomotion and focal adhesions are regulated by substrate flexibility. AB - Responses of cells to mechanical properties of the adhesion substrate were examined by culturing normal rat kidney epithelial and 3T3 fibroblastic cells on a collagen-coated polyacrylamide substrate that allows the flexibility to be varied while maintaining a constant chemical environment. Compared with cells on rigid substrates, those on flexible substrates showed reduced spreading and increased rates of motility or lamellipodial activity. Microinjection of fluorescent vinculin indicated that focal adhesions on flexible substrates were irregularly shaped and highly dynamic whereas those on firm substrates had a normal morphology and were much more stable. Cells on flexible substrates also contained a reduced amount of phosphotyrosine at adhesion sites. Treatment of these cells with phenylarsine oxide, a tyrosine phosphatase inhibitor, induced the formation of normal, stable focal adhesions similar to those on firm substrates. Conversely, treatment of cells on firm substrates with myosin inhibitors 2,3-butanedione monoxime or KT5926 caused the reduction of both vinculin and phosphotyrosine at adhesion sites. These results demonstrate the ability of cells to survey the mechanical properties of their surrounding environment and suggest the possible involvement of both protein tyrosine phosphorylation and myosin-generated cortical forces in this process. Such response to physical parameters likely represents an important mechanism of cellular interaction with the surrounding environment within a complex organism. PMID- 9391083 TI - Platelet-derived growth factor activates mitogen-activated protein kinase in isolated caveolae. AB - The ability of a peptide hormone to affect many different intracellular targets is thought to be possible because of the modular organization of signal transducing molecules in the cell. Evidence for the presence of signaling modules in metazoan cells, however, is incomplete. Herein we show, with morphology and cell fractionation, that all the components of a mitogen-activated protein kinase pathway are concentrated in caveolae of unstimulated human fibroblasts. Addition of platelet-derived growth factor to either the intact cell or caveolae isolated from these cells stimulates tyrosine phosphorylation and activates mitogen activated protein kinases in caveolae. The molecular machinery for kinase activation, therefore, is preorganized at the cell surface of quiescent cells. PMID- 9391085 TI - Telomerase activity: a biomarker of cell proliferation, not malignant transformation. AB - Telomerase activity is readily detected in most cancer biopsies, but not in premalignant lesions or in normal tissue samples with a few exceptions that include germ cells and hemopoietic stem cells. Telomerase activity may, therefore, be a useful biomarker for diagnosis of malignancies and a target for inactivation in chemotherapy or gene therapy. These observations have led to the hypothesis that activation of telomerase may be an important step in tumorigenesis. To test this hypothesis, we studied telomerase activity in isogeneic samples of uncultured and cultured specimens of normal human uroepithelial cells (HUCs) and in uncultured and cultured biopsies of superficial and myoinvasive transitional cell carcinoma (TCC) of the bladder. Our results demonstrated that four of four TCC biopsies, representing both superficial and myoinvasive TCCs, were positive for telomerase activity, but all samples of uncultured HUC were telomerase negative. However, when the same normal HUC samples were established as proliferating cultures in vitro, telomerase activity was readily detected but usually at lower levels than in TCCs. Consistent with the above observation of the telomerase activity in HUCs, telomeres did not shorten during the HUC in vitro lifespan. Demonstration of telomerase in proliferating human epithelial cells in vitro was not restricted to HUCs, because it was also present in prostate and mammary cell cultures. Notably, telomerase activity was relatively low or undetectable in nonproliferating HUC cultures. These data do not support a model in which telomerase is inactive in normal cells and activated during tumorigenic transformation. Rather, these data support a model in which the detection of telomerase in TCC biopsies, but not uncultured HUC samples, reflects differences in proliferation between tumor and normal cells in vivo. PMID- 9391084 TI - Mannose-6-phosphate/insulin-like growth factor-II receptor is a receptor for retinoic acid. AB - Retinoic acid (RA) exerts diverse biological effects in the control of cell growth in embryogenesis and oncogenesis. These effects of RA are thought to be mediated by the nuclear retinoid receptors. Mannose-6-phosphate (M6P)/insulin like growth factor-II (IGF-II) receptor is a multifunctional membrane glycoprotein that is known to bind both M6P and IGF-II and function primarily in the binding and trafficking of lysosomal enzymes, the activation of transforming growth factor-beta, and the degradation of IGF-II. M6P/IGF-II receptor has recently been implicated in fetal development and carcinogenesis. Despite the functional similarities between RA and the M6P/IGF-II receptor, no direct biochemical link has been established. Here, we show that the M6P/IGF-II receptor also binds RA with high affinity at a site that is distinct from those for M6P and IGF-II, as identified by a photoaffinity labeling technique. We also show that the binding of RA to the M6P/IGF-II receptor enhances the primary functions of this receptor. The biological consequence of the interaction appears to be the suppression of cell proliferation and/or induction of apoptosis. These findings suggest that the M6P/IGF-II receptor mediates a RA response pathway that is important in cell growth regulation. This discovery of the interaction of RA with the M6P/IGF-II receptor may have important implications for our understanding of the roles of RA and the M6P/IGF-II receptor in development, carcinogenesis, and lysosomal enzyme-related diseases. PMID- 9391086 TI - Syntenin, a PDZ protein that binds syndecan cytoplasmic domains. AB - The syndecans are transmembrane proteoglycans that place structurally heterogeneous heparan sulfate chains at the cell surface and a highly conserved polypeptide in the cytoplasm. Their versatile heparan sulfate moieties support various processes of molecular recognition, signaling, and trafficking. Here we report the identification of a protein that binds to the cytoplasmic domains of the syndecans in yeast two-hybrid screens, surface plasmon resonance experiments, and ligand-overlay assays. This protein, syntenin, contains a tandem repeat of PDZ domains that reacts with the FYA C-terminal amino acid sequence of the syndecans. Recombinant enhanced green fluorescent protein (eGFP)-syntenin fusion proteins decorate the plasmamembrane and intracellular vesicles, where they colocalize and cosegregate with syndecans. Cells that overexpress eGFP-syntenin show numerous cell surface extensions, suggesting effects of syntenin on cytoskeleton-membrane organization. We propose that syntenin may function as an adaptor that couples syndecans to cytoskeletal proteins or cytosolic downstream signal-effectors. PMID- 9391087 TI - Targeted expression of constitutively active receptors for parathyroid hormone and parathyroid hormone-related peptide delays endochondral bone formation and rescues mice that lack parathyroid hormone-related peptide. AB - Mice in which the genes encoding the parathyroid hormone (PTH)-related peptide (PTHrP) or the PTH/PTHrP receptor have been ablated by homologous recombination show skeletal dysplasia due to accelerated endochondral bone formation, and die at birth or in utero, respectively. Skeletal abnormalities due to decelerated chondrocyte maturation are observed in transgenic mice where PTHrP expression is targeted to the growth plate, and in patients with Jansen metaphyseal chondrodysplasia, a rare genetic disorder caused by constitutively active PTH/PTHrP receptors. These and other findings thus indicate that PTHrP and its receptor are essential for chondrocyte differentiation. To further explore the role of the PTH/PTHrP receptor in this process, we generated transgenic mice in which expression of a constitutively active receptor, HKrk-H223R, was targeted to the growth plate by the rat alpha1 (II) collagen promoter. Two major goals were pursued: (i) to investigate how constitutively active PTH/PTHrP receptors affect the program of chondrocyte maturation; and (ii) to determine whether expression of the mutant receptor would correct the severe growth plate abnormalities of PTHrP-ablated mice (PTHrP-/-). The targeted expression of constitutively active PTH/PTHrP receptors led to delayed mineralization, decelerated conversion of proliferative chondrocytes into hypertrophic cells in skeletal segments that are formed by the endochondral process, and prolonged presence of hypertrophic chondrocytes with delay of vascular invasion. Furthermore, it corrected at birth the growth plate abnormalities of PTHrP-/- mice and allowed their prolonged survival. "Rescued" animals lacked tooth eruption and showed premature epiphyseal closure, indicating that both processes involve PTHrP. These findings suggest that rescued PTHrP-/- mice may gain considerable importance for studying the diverse, possibly tissue-specific role(s) of PTHrP in postnatal development. PMID- 9391088 TI - Regulation of number and size of digits by posterior Hox genes: a dose-dependent mechanism with potential evolutionary implications. AB - The proper development of digits, in tetrapods, requires the activity of several genes of the HoxA and HoxD homeobox gene complexes. By using a variety of loss-of function alleles involving the five Hox genes that have been described to affect digit patterning, we report here that the group 11, 12, and 13 genes control both the size and number of murine digits in a dose-dependent fashion, rather than through a Hox code involving differential qualitative functions. A similar dose response is observed in the morphogenesis of the penian bone, the baculum, which further suggests that digits and external genitalia share this genetic control mechanism. A progressive reduction in the dose of Hox gene products led first to ectrodactyly, then to olygodactyly and adactyly. Interestingly, this transition between the pentadactyl to the adactyl formula went through a step of polydactyly. We propose that in the distal appendage of polydactylous short digited ancestral tetrapods, such as Acanthostega, the HoxA complex was predominantly active. Subsequent recruitment of the HoxD complex contributed to both reductions in digit number and increase in digit length. Thus, transition through a polydactylous limb before reaching and stabilizing the pentadactyl pattern may have relied, at least in part, on asynchronous and independent changes in the regulation of HoxA and HoxD gene complexes. PMID- 9391089 TI - Conservation of fibroblast growth factor function in lens regeneration. AB - In urodele amphibians, lens induction during development and regeneration occurs through different pathways. During development, the lens is induced from the mutual interaction of the ectoderm and the optic vesicle, whereas after lentectomy the lens is regenerated through the transdifferentiation of the iris pigmented epithelial cells. Given the known role of fibroblast growth factors (FGFs) during lens development, we examined whether or not the expression and the effects of exogenous FGF during urodele lens regeneration were conserved. In this paper, we describe expression of FGF-1 and its receptors, FGFR-2 (KGFR and bek variants) and FGFR-3, in newts during lens regeneration. Expression of these genes was readily observed in the dedifferentiating pigmented epithelial cells, and the levels of expression were high in the lens epithelium and the differentiating fibers and lower in the retina. These patterns of expression implied involvement of FGFs in lens regeneration. To further elucidate this function, we examined the effects of exogenous FGF-1 and FGF-4 during lens regeneration. FGF-1 or FGF-4 treatment in lentectomized eyes resulted in the induction of abnormalities reminiscent to the ones induced during lens development in transgenic mice. Effects included transformation of epithelial cells to fiber cells, double lens regeneration, and lenses with abnormal polarity. These results establish that FGF molecules are key factors in fiber differentiation, polarity, and morphogenesis of the lens during regeneration even though the regenerating lens is induced by a different mechanism than in lens development. In this sense, FGF function in lens regeneration and development should be regarded as conserved. Such conservation should help elucidate the mechanisms of lens regeneration in urodeles and its absence in higher vertebrates. PMID- 9391090 TI - The LIM-domain binding protein Ldb1 and its partner LMO2 act as negative regulators of erythroid differentiation. AB - The nuclear LIM domain protein LMO2, a T cell oncoprotein, is essential for embryonic erythropoiesis. LIM-only proteins are presumed to act primarily through protein-protein interactions. We, and others, have identified a widely expressed protein, Ldb1, whose C-terminal 76-residues are sufficient to mediate interaction with LMO2. In murine erythroleukemia cells, the endogenous Lbd1 and LMO2 proteins exist in a stable complex, whose binding affinity appears greater than that between LMO2 and the bHLH transcription factor SCL. However, Ldb1, LMO2, and SCL/E12 can assemble as a multiprotein complex on a consensus SCL binding site. Like LMO2, the Ldb1 gene is expressed in fetal liver and erythroid cell lines. Forced expression of Ldb1 in G1ER proerythroblast cells inhibited cellular maturation, a finding compatible with the decrease in Ldb1 gene expression that normally occurs during erythroid differentiation. Overexpression of the LMO2 gene also inhibited erythroid differentiation. Our studies demonstrate a function for Ldb1 in hemopoietic cells and suggest that one role of the Ldb1/LMO2 complex is to maintain erythroid precursors in an immature state. PMID- 9391092 TI - Selection for spiral waves in the social amoebae Dictyostelium. AB - Starving Dictyostelium amoebae emit pulses of the chemoattractant cAMP that are relayed from cell to cell as circular and spiral waves. We have recently modeled spiral wave formation in Dictyostelium. Our model suggests that a secreted protein inhibitor of an extracellular cAMP phosphodiesterase selects for spirals. Herein we test the essential features of this prediction by comparing wave propagation in wild type and inhibitor mutants. We find that mutants rarely form spirals. The territory size of mutant strains is approximately 50 times smaller than wild type, and the mature fruiting bodies are smaller but otherwise normal. These results identify a mechanism for selecting one wave symmetry over another in an excitable system and suggest that the phosphodiesterase inhibitor may be under selection because it helps regulate territory size. PMID- 9391091 TI - Regulation of dorsal fate in the neuraxis by Wnt-1 and Wnt-3a. AB - Members of the Wnt family of signaling molecules are expressed differentially along the dorsal-ventral axis of the developing neural tube. Thus we asked whether Wnt factors are involved in patterning of the nervous system along this axis. We show that Wnt-1 and Wnt-3a, both of which are expressed in the dorsal portion of the neural tube, could synergize with the neural inducers noggin and chordin in Xenopus animal explants to generate the most dorsal neural structure, the neural crest, as determined by the expression of Krox-20, AP-2, and slug. Overexpression of Wnt-1 or Wnt-3a in the neuroectoderm of whole embryos led to a dramatic increase of slug and Krox-20-expressing cells, but the hindbrain expression of Krox-20 remained unaffected. Enlargement in the neural crest population could occur even when cell proliferation was inhibited. Wnt-5A and Wnt 8, neither of which is expressed in the dorsal neuroectoderm, failed to induce neural crest markers. Overexpression of glycogen synthase kinase 3, known to antagonize Wnt signaling, blocked the neural-crest-inducing activity of Wnt-3a in animal explants and inhibited neural crest formation in whole embryos. We suggest that Wnt-1 and Wnt-3a have a role in patterning the neural tube along its dorsoventral axis and function in the differentiation of the neural crest. PMID- 9391094 TI - From tropics to tundra: global convergence in plant functioning. AB - Despite striking differences in climate, soils, and evolutionary history among diverse biomes ranging from tropical and temperate forests to alpine tundra and desert, we found similar interspecific relationships among leaf structure and function and plant growth in all biomes. Our results thus demonstrate convergent evolution and global generality in plant functioning, despite the enormous diversity of plant species and biomes. For 280 plant species from two global data sets, we found that potential carbon gain (photosynthesis) and carbon loss (respiration) increase in similar proportion with decreasing leaf life-span, increasing leaf nitrogen concentration, and increasing leaf surface area-to-mass ratio. Productivity of individual plants and of leaves in vegetation canopies also changes in constant proportion to leaf life-span and surface area-to-mass ratio. These global plant functional relationships have significant implications for global scale modeling of vegetation-atmosphere CO2 exchange. PMID- 9391093 TI - Methylation of the minimal promoter of an embryonic globin gene silences transcription in primary erythroid cells. AB - Methylation of cytosines in the dinucleotide CpG has been shown to suppress transcription of a number of tissue-specific genes, yet the precise mechanism is not fully understood. The vertebrate globin genes were among the first examples in which an inverse correlation was shown between CpG methylation and transcription. We studied the methylation pattern of the 235-bp rho-globin gene promoter in genomic DNA from primary chicken erythroid cells using the sodium bisulfite conversion technique and found all CpGs in the promoter to be methylated in erythroid cells from adult chickens in which the rho-globin gene is silent but unmethylated in 5-day (primitive) embryonic red cells in which the gene is transcribed. To elucidate further the mechanism of methylation-induced silencing, an expression construct consisting of 235 bp of 5' promoter sequence of the rho-globin gene along with a strong 5' erythroid enhancer driving a chloramphenicol acetyltransferase reporter gene, rho-CAT, was transfected into primary avian erythroid cells derived from 5-day embryos. Methylation of just the 235-bp rho-globin gene promoter fragment at every CpG resulted in a 20- to 30 fold inhibition of transcription, and this effect was not overridden by the presence of potent erythroid-specific enhancers. The ability of the 235-bp rho globin gene promoter to bind to a DNA Methyl Cytosine binding Protein Complex (MeCPC) was tested in electrophoretic mobility shift assays utilizing primary avian erythroid cell nuclear extract. The results were that fully methylated but not unmethylated 235-bp rho-globin gene promoter fragment could compete efficiently for MeCPC binding. These results are a direct demonstration that site specific methylation of a globin gene promoter at the exact CpGs that are methylated in vivo can silence transcription in homologous primary erythroid cells. Further, these data implicate binding of MeCPC to the promoter in the mechanism of silencing. PMID- 9391095 TI - Ambient UV-B radiation causes deformities in amphibian embryos. AB - There has been a great deal of recent attention on the suspected increase in amphibian deformities. However, most reports of amphibian deformities have been anecdotal, and no experiments in the field under natural conditions have been performed to investigate this phenomenon. Under laboratory conditions, a variety of agents can induce deformities in amphibians. We investigated one of these agents, UV-B radiation, in field experiments, as a cause for amphibian deformities. We monitored hatching success and development in long-toed salamanders under UV-B shields and in regimes that allowed UV-B radiation. Embryos under UV-B shields had a significantly higher hatching rate and fewer deformities, and developed more quickly than those exposed to UV-B. Deformities may contribute directly to embryo mortality, and they may affect an individual's subsequent survival after hatching. PMID- 9391096 TI - Evolutionary specialization of the nuclear targeting apparatus. AB - The alpha- and beta-karyopherins (Kaps), also called importins, mediate the nuclear transport of proteins. All alpha-Kaps contain a central domain composed of eight approximately 40 amino acid, tandemly arranged, armadillo-like (Arm) repeats. The number and order of these repeats have not changed since the common origin of fungi, plants, and mammals. Phylogenetic analysis suggests that the various alpha-Kaps fall into two groups, alpha1 and alpha2. Whereas animals encode both types, the yeast genome encodes only an alpha1-Kap. The beta-Kaps are characterized by 14-15 tandemly arranged HEAT motifs. We show that the Arm repeats of alpha-Kaps and the HEAT motifs of beta-Kaps are similar, suggesting that the alpha-Kaps and beta-Kaps (and for that matter, all Arm and HEAT repeat containing proteins) are members of the same protein superfamily. Phylogenetic analysis indicates that there are at least three major groups of beta-Kaps, consistent with their proposed cargo specificities. We present a model in which an alpha-independent beta-Kap progenitor gave rise to the alpha-dependent beta Kaps and the alpha-Kaps. PMID- 9391098 TI - Did homeodomain proteins duplicate before the origin of angiosperms, fungi, and metazoa? AB - Homeodomain proteins are transcription factors that play a critical role in early development in eukaryotes. These proteins previously have been classified into numerous subgroups whose phylogenetic relationships are unclear. Our phylogenetic analysis of representative eukaryotic sequences suggests that there are two major groups of homeodomain proteins, each containing sequences from angiosperms, metazoa, and fungi. This result, based on parsimony and neighbor-joining analyses of primary amino acid sequences, was supported by two additional features of the proteins. The two protein groups are distinguished by an insertion/deletion in the homeodomain, between helices I and II. In addition, an amphipathic alpha helical secondary structure in the region N terminal of the homeodomain is shared by angiosperm and metazoan sequences in one group. These results support the hypothesis that there was at least one duplication of homeobox genes before the origin of angiosperms, fungi, and metazoa. This duplication, in turn, suggests that these proteins had diverse functions early in the evolution of eukaryotes. The shared secondary structure in angiosperm and metazoan sequences points to an ancient conserved functional domain. PMID- 9391097 TI - Molecular evolution of two vertebrate aryl hydrocarbon (dioxin) receptors (AHR1 and AHR2) and the PAS family. AB - The aryl hydrocarbon receptor (AHR) is a ligand-activated transcription factor through which halogenated aromatic hydrocarbons such as 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) cause altered gene expression and toxicity. The AHR belongs to the basic helix-loop-helix/Per-ARNT-Sim (bHLH-PAS) family of transcriptional regulatory proteins, whose members play key roles in development, circadian rhythmicity, and environmental homeostasis; however, the normal cellular function of the AHR is not yet known. As part of a phylogenetic approach to understanding the function and evolutionary origin of the AHR, we sequenced the PAS homology domain of AHRs from several species of early vertebrates and performed phylogenetic analyses of these AHR amino acid sequences in relation to mammalian AHRs and 24 other members of the PAS family. AHR sequences were identified in a teleost (the killifish Fundulus heteroclitus), two elasmobranch species (the skate Raja erinacea and the dogfish Mustelus canis), and a jawless fish (the lamprey Petromyzon marinus). Two putative AHR genes, designated AHR1 and AHR2, were found both in Fundulus and Mustelus. Phylogenetic analyses indicate that the AHR2 genes in these two species are orthologous, suggesting that an AHR gene duplication occurred early in vertebrate evolution and that multiple AHR genes may be present in other vertebrates. Database searches and phylogenetic analyses identified four putative PAS proteins in the nematode Caenorhabditis elegans, including possible AHR and ARNT homologs. Phylogenetic analysis of the PAS gene family reveals distinct clades containing both invertebrate and vertebrate PAS family members; the latter include paralogous sequences that we propose have arisen by gene duplication early in vertebrate evolution. Overall, our analyses indicate that the AHR is a phylogenetically ancient protein present in all living vertebrate groups (with a possible invertebrate homolog), thus providing an evolutionary perspective to the study of dioxin toxicity and AHR function. PMID- 9391099 TI - The interphotoreceptor retinoid binding protein gene in therian mammals: implications for higher level relationships and evidence for loss of function in the marsupial mole. AB - The subclass Theria of Mammalia includes marsupials (infraclass Metatheria) and placentals (infraclass Eutheria). Within each group, interordinal relationships remain unclear. One limitation of many studies is incomplete ordinal representation. Here, we analyze DNA sequences for part of exon 1 of the interphotoreceptor retinoid binding protein gene, including 10 that are newly reported, for representatives of all therian orders. Among placentals, the most robust clades are Cetartiodactyla, Paenungulata, and an expanded African clade that includes paenungulates, tubulidentates, and macroscelideans. Anagalida, Archonta, Altungulata, Hyracoidea + Perissodactyla, Ungulata, and the "flying primate" hypothesis are rejected by statistical tests. Among marsupials, the most robust clade includes all orders except Didelphimorphia. The phylogenetic placement of the monito del monte and the marsupial mole remains unclear. However, the marsupial mole sequence contains three frameshift indels and numerous stop codons in all three reading frames. Given that the interphotoreceptor retinoid binding protein gene is a single-copy gene that functions in the visual cycle and that the marsupial mole is blind with degenerate eyes, this finding suggests that phenotypic degeneration of the eyes is accompanied by parallel changes at the molecular level as a result of relaxed selective constraints. PMID- 9391100 TI - Dentition of Proteopithecus sylviae, an archaic anthropoid from the Fayum, Egypt. AB - Proteopithecus sylviae is an archaic anthropoid from the late Eocene quarry L-41, Fayum Province, Egypt. The dentition of Proteopithecus is very primitive and does not closely resemble that of other, better known, primates from the Fayum (e.g., parapithecids and propliopithecids). The dental morphology, much of which is described herein, shows a platyrrhine-like level of organization, suggesting that P. sylviae may occupy a position near the base of the modern anthropoid radiation. PMID- 9391101 TI - Transcription factor Mts1/Mts2 (Atf1/Pcr1, Gad7/Pcr1) activates the M26 meiotic recombination hotspot in Schizosaccharomyces pombe. AB - Homologous recombination hotspots increase the frequency of recombination in nearby DNA. The M26 hotspot in the ade6 gene of Schizosaccharomyces pombe is a meiotic hotspot with a discrete, cis-acting nucleotide sequence (5'-ATGACGT-3') defined by extensive mutagenesis. A heterodimeric M26 DNA binding protein, composed of subunits Mts1 and Mts2, has been identified and purified 40,000-fold. Cloning, disruption, and genetic analyses of the mts genes demonstrate that the Mts1/Mts2 heterodimer is essential for hotspot activity. This provides direct evidence that a specific trans-acting factor, binding to a cis-acting site with a unique nucleotide sequence, is required to activate this meiotic hotspot. Intriguingly, the Mts1/Mts2 protein subunits are identical to the recently described transcription factors Atf1 (Gad7) and Pcr1, which are required for a variety of stress responses. However, we report differential dependence on the Mts proteins for hotspot activation and stress response, suggesting that these proteins are multifunctional and have distinct activities. Furthermore, ade6 mRNA levels are equivalent in hotspot and nonhotspot meioses and do not change in mts mutants, indicating that hotspot activation is not a consequence of elevated transcription levels. These findings suggest an intimate but separable link between the regulation of transcription and meiotic recombination. Other studies have recently shown that the Mts1/Mts2 protein and M26 sites are involved in meiotic recombination elsewhere in the S. pombe genome, suggesting that these factors help regulate the timing and distribution of homologous recombination. PMID- 9391102 TI - Replacement of Fhit in cancer cells suppresses tumorigenicity. AB - The candidate tumor suppressor gene, FHIT, encompasses the common human chromosomal fragile site at 3p14.2, the hereditary renal cancer translocation breakpoint, and cancer cell homozygous deletions. Fhit hydrolyzes dinucleotide 5',5"'-P1,P3-triphosphate in vitro and mutation of a central histidine abolishes hydrolase activity. To study Fhit function, wild-type and mutant FHIT genes were transfected into cancer cell lines that lacked endogenous Fhit. No consistent effect of exogenous Fhit on growth in culture was observed, but Fhit and hydrolase "dead" Fhit mutant proteins suppressed tumorigenicity in nude mice, indicating that 5',5"'-P1, P3-triphosphate hydrolysis is not required for tumor suppression. PMID- 9391103 TI - A ribozyme-mediated, gene "knockdown" strategy for the identification of gene function in zebrafish. AB - The zebrafish system offers many unique opportunities for the study of molecular biology. To date, only random mutagenesis, and not directed gene knockouts, have been demonstrated in this system. To more fully develop the potential of the zebrafish system, an approach to effectively inhibit the expression of any targeted gene in the developing zebrafish embryo has been developed. This approach uses a transient, cytoplasmic, T7 expression system, injected into the fertilized zebrafish egg to rapidly produce high levels of a ribozyme directed against the mRNA encoded by the targeted gene to inhibit its expression. In a demonstration of this strategy, expression of the recessive dominant zebrafish no tail gene was effectively inhibited by using this strategy to yield a phenotype identical to that resulting from a known defective mutation in this same gene. This, ribozyme-mediated, message deletion strategy may have use in determining the function of genetic coding sequences of unknown function. PMID- 9391104 TI - In vivo telomere dynamics of human hematopoietic stem cells. AB - Aging in vivo and cell division in vitro are associated with telomere shortening. Several lines of evidence suggest that telomere length may be a good predictor of the long term replicative capacity of cells. To investigate the natural fate of chromosome telomeres of hematopoietic stem cells in vivo, we measured the telomere length of peripheral blood granulocytes from 11 fully engrafted bone marrow transplant recipients and from their respective donors. In 10 of 11 donor recipient pairs, the telomere length was significantly reduced in the recipient and the extent of reduction correlated inversely with the number of nucleated cells infused. These data provide internally controlled in vivo evidence that, concomitantly with their proliferation, hematopoietic stem cells lose telomere length; it is possible that, as a result, their proliferative potential is reduced. These findings must be taken into account when developing new protocols in which few stem cells are used for bone marrow transplantation or for gene therapy. PMID- 9391105 TI - Genetic recombination of poliovirus in a cell-free system. AB - Genetic recombination of plus-strand RNA viruses is an important process for promoting genetic variation. By using genetically marked poliovirus RNAs, we have demonstrated that genetic recombination can occur in a cell-free system that generates infective virus from added poliovirus RNA. Recombinant polioviruses were isolated, and the region of crossing over was roughly mapped. Recombinants could be isolated even under conditions where the yield of viruses from one of the parental RNAs was depressed to levels comparable to or less than the yield of recombinant viruses, an observation suggesting that only one of the recombining RNAs needs to be replication-competent. The generation of poliovirus recombinants in a cell-free system offers new possibilities for studying recombination and evolution of RNA viruses. PMID- 9391106 TI - Multiple pathways for SOS-induced mutagenesis in Escherichia coli: an overexpression of dinB/dinP results in strongly enhancing mutagenesis in the absence of any exogenous treatment to damage DNA. AB - dinP is an Escherichia coli gene recently identified at 5.5 min of the genetic map, whose product shows a similarity in amino acid sequence to the E. coli UmuC protein involved in DNA damage-induced mutagenesis. In this paper we show that the gene is identical to dinB, an SOS gene previously localized near the lac locus at 8 min, the function of which was shown to be required for mutagenesis of nonirradiated lambda phage infecting UV-preirradiated bacterial cells (termed lambdaUTM for lambda untargeted mutagenesis). A newly constructed dinP null mutant exhibited the same defect for lambdaUTM as observed previously with a dinB::Mu mutant, and the defect was complemented by plasmids carrying dinP as the only intact bacterial gene. Furthermore, merely increasing the dinP gene expression, without UV irradiation or any other DNA-damaging treatment, resulted in a strong enhancement of mutagenesis in F'lac plasmids; at most, 800-fold increase in the G6-to-G5 change. The enhanced mutagenesis did not depend on recA, uvrA, or umuDC. Thus, our results establish that E. coli has at least two distinct pathways for SOS-induced mutagenesis: one dependent on umuDC and the other on dinB/P. PMID- 9391107 TI - Mitochondrial mutational spectra in human cells and tissues. AB - We have found that human organs such as colon, lung, and muscle, as well as their derived tumors, share nearly all mitochondrial hotspot point mutations. Seventeen hotspots, primarily G --> A and A --> G transitions, have been identified in the mitochondrial sequence of base pairs 10,030-10,130. Mutant fractions increase with the number of cell generations in a human B cell line, TK6, indicating that they are heritable changes. The mitochondrial point mutation rate appears to be more than two orders of magnitude higher than the nuclear point mutation rate in TK6 cells and in human tissues. The similarity of the hotspot sets in vivo and in vitro leads us to conclude that human mitochondrial point mutations in the sequence studied are primarily spontaneous in origin and arise either from DNA replication error or reactions of DNA with endogenous metabolites. The predominance of transition mutations and the high number of hotspots in this short sequence resembles spectra produced by DNA polymerases in vitro. PMID- 9391109 TI - Yeast mutations in multiple complementation groups inhibit brome mosaic virus RNA replication and transcription and perturb regulated expression of the viral polymerase-like gene. AB - Brome mosaic virus (BMV), a member of the alphavirus-like superfamily of positive strand RNA viruses, encodes two proteins, 1a and 2a, that interact with each other, with unidentified host proteins, and with host membranes to form the viral RNA replication complex. Yeast expressing 1a and 2a support replication and subgenomic mRNA synthesis by BMV RNA3 derivatives. Using a multistep selection and screening process, we have isolated yeast mutants in multiple complementation groups that inhibit BMV-directed gene expression. Three complementation groups, represented by mutants mab1-1, mab2-1, and mab3-1 (for maintenance of BMV functions), were selected for initial study. Each of these mutants has a single, recessive, chromosomal mutation that inhibits accumulation of positive- and negative-strand RNA3 and subgenomic mRNA. BMV-directed gene expression was inhibited when the RNA replication template was introduced by in vivo transcription from DNA or by transfection of yeast with in vitro transcripts, confirming that cytoplasmic RNA replication steps were defective. mab1-1, mab2-1, and mab3-1 slowed yeast growth to varying degrees and were temperature-sensitive, showing that the affected genes contribute to normal cell growth. In wild-type yeast, expression of the helicase-like 1a protein increased the accumulation of 2a mRNA and the polymerase-like 2a protein, revealing a new level of viral regulation. In association with their other effects, mab1-1 and mab2-1 blocked the ability of 1a to stimulate 2a mRNA and protein accumulation, whereas mab3-1 had elevated 2a protein accumulation. Together, these results show that BMV RNA replication in yeast depends on multiple host genes, some of which directly or indirectly affect the regulated expression and accumulation of 2a. PMID- 9391108 TI - A family of genes required for maintenance of cell wall integrity and for the stress response in Saccharomyces cerevisiae. AB - The PKC1-MPK1 pathway in yeast functions in the maintenance of cell wall integrity and in the stress response. We have identified a family of genes that are putative regulators of this pathway. WSC1, WSC2, and WSC3 encode predicted integral membrane proteins with a conserved cysteine motif and a WSC1-green fluorescence protein fusion protein localizes to the plasma membrane. Deletion of WSC results in phenotypes similar to mutants in the PKC1-MPK1 pathway and an increase in the activity of MPK1 upon a mild heat treatment is impaired in a wscDelta mutant. Genetic analysis places the function of WSC upstream of PKC1, suggesting that they play a role in its activation. We also find a genetic interaction between WSC and the RAS-cAMP pathway. The RAS-cAMP pathway is required for cell cycle progression and for the heat shock response. Overexpression of WSC suppresses the heat shock sensitivity of a strain in which RAS is hyperactivated and the heat shock sensitivity of a wscDelta strain is rescued by deletion of RAS2. The functional characteristics and cellular localization of WSC suggest that they may mediate intracellular responses to environmental stress in yeast. PMID- 9391110 TI - Ascorbate recycling in human neutrophils: induction by bacteria. AB - Ascorbate (vitamin C) recycling occurs when extracellular ascorbate is oxidized, transported as dehydroascorbic acid, and reduced intracellularly to ascorbate. We investigated microorganism induction of ascorbate recycling in human neutrophils and in microorganisms themselves. Ascorbate recycling was determined by measuring intracellular ascorbate accumulation. Ascorbate recycling in neutrophils was induced by both Gram-positive and Gram-negative pathogenic bacteria, and the fungal pathogen Candida albicans. Induction of recycling resulted in as high as a 30-fold increase in intracellular ascorbate compared with neutrophils not exposed to microorganisms. Recycling occurred at physiologic concentrations of extracellular ascorbate within 20 min, occurred over a 100-fold range of effector/target ratios, and depended on oxidation of extracellular ascorbate to dehydroascorbic acid. Ascorbate recycling did not occur in bacteria nor in C. albicans. Ascorbate did not enter microorganisms, and dehydroascorbic acid entry was less than could be accounted for by diffusion. Because microorganism lysates reduced dehydroascorbic acid to ascorbate, ascorbate recycling was absent because of negligible entry of the substrate dehydroascorbic acid. Because ascorbate recycling occurs in human neutrophils but not in microorganisms, it may represent a eukaryotic defense mechanism against oxidants with possible clinical implications. PMID- 9391111 TI - Phosphatidylinositol 3-kinase-gamma activates Bruton's tyrosine kinase in concert with Src family kinases. AB - Bruton's tyrosine kinase (Btk) is essential for normal B lymphocyte development and function. The activity of Btk is partially regulated by transphosphorylation within its kinase domain by Src family kinases at residue Tyr-551 and subsequent autophosphorylation at Tyr-223. Activation correlates with Btk association with cellular membranes. Based on specific loss of function mutations, the Btk pleckstrin homology (PH) domain plays an essential role in this activation process. The Btk PH domain can bind in vitro to several lipid end products of the phosphatidylinositol 3-kinase (PI 3-kinase) family including phosphatidylinositol 3,4,5-trisphosphate. Activation of Btk as monitored by elevation of phosphotyrosine content and a cellular transformation response was dramatically enhanced by coexpressing a weakly activated allele of Src (E378G) and the two subunits of PI 3-kinase-gamma. This activation correlates with new sites of phosphorylation on Btk identified by two-dimensional phosphopeptide mapping. Activation of Btk was dependent on the catalytic activity of all three enzymes and an intact Btk PH domain and Src transphosphorylation site. These combined data define Btk as a downstream target of PI 3-kinase-gamma and Src family kinases. PMID- 9391112 TI - An in vitro study of the dynamic features of the major histocompatibility complex class I complex relevant to its role as a versatile peptide-receptive molecule. AB - The major histocompatibility complex class I complex consists of a heavy chain and a light chain (beta2-microglobulin, beta2m), which assemble with a short endogenously derived peptide in the endoplasmic reticulum. The class I peptide can be directly exchanged, either at the cell surface or, as recently described, in vesicles of the endocytic compartments, thus allowing exogenous peptides to enter the class I presentation pathway. To probe the interactions between the components of the class I molecule, we analyzed the exchange of peptide and beta2m by using purified, recombinant H2-Kb/peptide complexes in a cell-free in vitro system. The exchange of competitor peptide was primarily dependent on the off-rate of the original peptide in the class I binding groove. Peptide exchange was not enhanced by the presence of exogenous beta2m, as exchange occurred to the same extent in its absence. Thus, the exchange of peptide and beta2m are independent events. The exchange rate of beta2m also was not affected by the dissociation rates of the original peptides. Furthermore, peptides could substantially exchange into class I molecules over a pH range of 5.5 to 7.5, conditions prevalent in certain endocytic compartments. We conclude that the dynamic properties of the components of class I molecules explain its function as a highly peptide-receptive molecule. The major histocompatibility complex class I can readily receive peptides independent of the presence of exogenous beta2m, even at a low pH. Such properties are relevant to class I peptide acquisition, which can occur at the cell surface, as well as in specialized endosomes. PMID- 9391113 TI - Decreased ability of HIV-1 tat protein-treated accessory cells to organize cellular clusters is associated with partial activation of T cells. AB - It has been shown in several animal models that HIV infection of accessory cells (ACs) plays an important role in development of AIDS. Here, we report that ACs treated with HIV-1 Tat protein (Tat-ACs) have a decreased ability to organize cellular aggregates as compared with untreated ACs, resulting in incomplete activation of T cells in responses to anti-CD3 mAb or staphylococcal enterotoxin B stimulation. The T cells failed to up-regulate adhesion molecules CD11a and CD2 on the cell surface and had reduced proliferative responses, as determined by [3H]thymidine incorporation, but they obtained lymphoblast-like morphology and expressed early activation antigens on the cell surface such as Fas and CD69 and interleukin 2 receptor, at comparable levels as those T cells undergoing a maximal proliferation. These results suggest that the Tat-AC-induced defect occurs in the late, but not in the early, phases of T cell activation. Normal expression of cell surface Fas antigen accompanied by defects in late activation thus may result in the susceptibility of these T cells to apoptosis. Our studies suggest that dysfunction, hyperactivation, and susceptibility to apoptosis, as observed with T cells isolated from HIV-infected individuals, may be, at least in part, a consequence of abnormal functions of ACs. PMID- 9391114 TI - Alphabeta T cell receptor interactions with syngeneic and allogeneic ligands: affinity measurements and crystallization. AB - Cellular immunity is mediated by the interaction of an alphabeta T cell receptor (TCR) with a peptide presented within the context of a major histocompatibility complex (MHC) molecule. Alloreactive T cells have alphabeta TCRs that can recognize both self- and foreign peptide-MHC (pMHC) complexes, implying that the TCR has significant complementarity with different pMHC. To characterize the molecular basis for alloreactive TCR recognition of pMHC, we have produced a soluble, recombinant form of an alloreactive alphabeta T cell receptor in Drosophila melanogaster cells. This recombinant TCR, 2C, is expressed as a correctly paired alphabeta heterodimer, with the chains covalently connected via a disulfide bond in the C-terminal region. The native conformation of the 2C TCR was probed by surface plasmon resonance (SPR) analysis by using conformation specific monoclonal antibodies, as well as syngeneic and allogeneic pMHC ligands. The 2C interaction with H-2Kb-dEV8, H-2Kbm3-dEV8, H-2Kb-SIYR, and H-2Ld-p2Ca spans a range of affinities from Kd = 10(-4) to 10(-6)M for the syngeneic (H-2Kb) and allogeneic (H-2Kbm3, H-2Ld) ligands. In general, the syngeneic ligands bind with weaker affinities than the allogeneic ligands, consistent with current threshold models of thymic selection and T cell activation. Crystallization of the 2C TCR required proteolytic trimming of the C-terminal residues of the alpha and beta chains. X-ray quality crystals of complexes of 2C with H-2Kb-dEV8, H 2Kbm3-dEV8 and H-2Kb-SIYR have been grown. PMID- 9391115 TI - Interferon-gamma impacts at multiple points during the progression of autoimmune diabetes. AB - The role of interferon-gamma in autoimmune diabetes was assessed by breeding a null mutation of the interferon-gamma receptor alpha chain into the nonobese diabetic mouse strain, as well as into a simplified T cell receptor transgenic model of diabetes. In contrast to a previous report on abrogation of the interferon-gamma gene, mutation of the gene encoding its receptor led to drastic effects on disease in both mouse lines. Nonobese diabetic mice showed a marked inhibition of insulitis-both the kinetics and penetrance-and no signs of diabetes; the transgenic model exhibited near-normal insulitis, but this never evolved into diabetes, either spontaneously or after experimental provocation. This failure could not be explained by perturbations in the ratio of T helper cell phenotypes; rather, it reflected a defect in antigen-presenting cells or in the islet beta cell targets. PMID- 9391116 TI - Distinct tyrosine phosphorylation sites in JAK3 kinase domain positively and negatively regulate its enzymatic activity. AB - Cytokines are critically important for the growth and development of a variety of cells. Janus kinases (JAKs) associate with cytokine receptors and are essential for transmitting downstream cytokine signals. However, the regulation of the enzymatic activity of the JAKs is not well understood. Here, we investigated the role of tyrosine phosphorylation of JAK3 in regulating its kinase activity by analyzing mutations of tyrosine residues within the putative activation loop of the kinase domain. Specifically, tyrosine residues 980 and 981 of JAK3 were mutated to phenylalanine individually or doubly. We found that JAK3 is autophosphorylated on multiple sites including Y980 and Y981. Compared with the activity of wild-type (WT) JAK3, mutant Y980F demonstrated markedly decreased kinase activity, and optimal phosphorylation of JAK3 on other sites was dependent on Y980 phosphorylation. The mutant Y980F also exhibited reduced phosphorylation of its substrates, gammac and STAT5A. In contrast, mutant Y981F had greatly increased kinase activity, whereas the double mutant, YY980/981FF, had intermediate activity. These results indicate that Y980 positively regulates JAK3 kinase activity whereas Y981 negatively regulates JAK3 kinase activity. These observations in JAK3 are similar to the findings in the kinase that is closely related to the JAK family, ZAP-70; mutations of tyrosine residues within the putative activation loop of ZAP-70 also have opposing actions. Thus, it will be important to determine whether this feature of regulation is unique to JAK3 or if it is also a feature of other JAKs. Given the importance of JAKs and particularly JAK3, it will be critical to fully dissect the positive and negative regulatory function of these and other tyrosine residues in the control of kinase activity and hence cytokine signaling. PMID- 9391117 TI - Production, specificity, and functionality of monoclonal antibodies to specific peptide-major histocompatibility complex class II complexes formed by processing of exogenous protein. AB - Several unanswered questions in T cell immunobiology relating to intracellular processing or in vivo antigen presentation could be approached if convenient, specific, and sensitive reagents were available for detecting the peptide-major histocompatibility complex (MHC) class I or class II ligands recognized by alphabeta T cell receptors. For this reason, we have developed a method using homogeneously loaded peptide-MHC class II complexes to generate and select specific mAb reactive with these structures using hen egg lysozyme (HEL) and I-Ak as a model system. mAbs specific for either HEL-(46-61)-Ak or HEL-(116-129)-Ak have been isolated. They cross-react with a small subset of I-Ak molecules loaded with self peptides but can nonetheless be used for flow cytometry, immunoprecipitation, Western blotting, and intracellular immunofluorescence to detect specific HEL peptide-MHC class II complexes formed by either peptide exposure or natural processing of native HEL. An example of the utility of these reagents is provided herein by using one of the anti-HEL-(46-61)-Ak specific mAbs to visualize intracellular compartments where I-Ak is loaded with HEL-derived peptides early after antigen administration. Other uses, especially for in vivo tracking of specific ligand-bearing antigen-presenting cells, are discussed. PMID- 9391118 TI - Gene transfer of Fas ligand induces tumor regression in vivo. AB - The Fas-Fas ligand (FasL) system plays an important role in the induction of lymphoid apoptosis and has been implicated in the suppression of immune responses. Herein, we report that gene transfer of FasL inhibits tumor cell growth in vivo. Although such inhibition is expected in Fas+ tumor cell lines, marked regression was unexpectedly observed after FasL gene transfer into the CT26 colon carcinoma that does not express Fas. Infection by an adenoviral vector encoding FasL rapidly eliminated tumor masses in the Fas+ Renca tumor by inducing cell death, whereas the elimination of Fas- CT26 cells was mediated by inflammatory cells. Analysis of human malignancies revealed Fas, but not FasL, expression in a majority of tumors and susceptibility to FasL in most Fas+ cell lines. These findings suggest that gene transfer of FasL generates apoptotic responses and induces potent inflammatory reactions that can be used to induce the regression of malignancies. PMID- 9391119 TI - A point mutation in the MET oncogene abrogates metastasis without affecting transformation. AB - The MET oncogene encodes the tyrosine kinase receptor for hepatocyte growth factor/scatter factor (HGF), known to stimulate invasive growth of epithelial cells. MET is overexpressed in a significant percentage of human cancers and is amplified during the transition between primary tumors and metastasis. To investigate whether this oncogene is directly responsible for the acquisition of the metastatic phenotype, we exploited a single-hit oncogenic version of MET, able to transform and to confer invasive and metastatic properties to nontumorigenic cells, both in vitro and in nude mice. We mutagenized the signal transducer docking site of Met (Y1349VHVX3Y1356VNV), which has the uncommon property of binding and activating multiple src homology region 2 (SH2) containing intracellular effectors. Notably, a point mutation (H1351 --> N) increased the transforming ability of the oncogene but abolished its metastatic potential. This mutation duplicates the Grb2 binding site, super-activating the Ras pathway and preventing the binding of the other intracellular transducers. Complementation in trans with another nonmetastatic mutant (N1358 --> H), recruiting all the transducers downstream to Met except Grb2, rescued the invasive-metastatic phenotype. It is concluded that the metastatic potential of the MET oncogene relies on the properties of its multifunctional docking site, and that a single point mutation affecting signal transduction can dissociate neoplastic transformation from metastasis. PMID- 9391120 TI - Both hypertrophic and dilated cardiomyopathies are caused by mutation of the same gene, delta-sarcoglycan, in hamster: an animal model of disrupted dystrophin associated glycoprotein complex. AB - Cardiomyopathy (CM) is a primary degenerative disease of myocardium and is traditionally categorized into hypertrophic and dilated CMs (HCM and DCM) according to its gross appearance. Cardiomyopathic hamster (CM hamster), a representative model of human hereditary CM, has HCM and DCM inbred sublines, both of which descend from the same ancestor. Herein we show that both HCM and DCM hamsters share a common defect in a gene for delta-sarcoglycan (delta-SG), the functional role of which is yet to be characterized. A breakpoint causing genomic deletion was found to be located at 6.1 kb 5' upstream of the second exon of delta-SG gene, and its 5' upstream region of more than 27.4 kb, including the authentic first exon of delta-SG gene, was deleted. This deletion included the major transcription initiation site, resulting in a deficiency of delta-SG transcripts with the consequent loss of delta-SG protein in all the CM hamsters, despite the fact that the protein coding region of delta-SG starting from the second exon was conserved in all the CM hamsters. We elucidated the molecular interaction of dystrophin-associated glycoproteins including delta-SG, by using an in vitro pull-down study and ligand overlay assay, which indicates the functional role of delta-SG in stabilizing sarcolemma. The present study not only identifies CM hamster as a valuable animal model for studying the function of delta-SG in vivo but also provides a genetic target for diagnosis and treatment of human CM. PMID- 9391121 TI - Identification, isolation, and characterization of daintain (allograft inflammatory factor 1), a macrophage polypeptide with effects on insulin secretion and abundantly present in the pancreas of prediabetic BB rats. AB - A bioactive macrophage factor, the polypeptide daintain/allograft inflammatory factor 1 (AIF1), has been isolated from porcine intestine. It was discovered when searching for intestinal peptides with effects on insulin release, and its purification was monitored by the influence of the peptide fractions on pancreatic glucose-induced insulin secretion. Daintain/AIF1 is a 146-aa residue polypeptide with a mass of 16,603 Da and an acetylated N terminus. An internal 44 residue segment with the sequence pattern -KR-KK-GKR- has a motif typical of peptide hormone precursors, i.e., dibasic sites for potential activation cleavages and at the sequentially last such site, the structure GKR. The latter is a signal for C-terminal amide formation in the processing of peptide hormones. Daintain/AIF1 is immunohistochemically localized to microglial cells in the central nervous system and to dendritic cells and macrophages in several organs. A particularly dense accumulation of daintain/AIF1-immunoreactive macrophages was observed in the insulitis affecting the pancreatic islets of prediabetic BB rats. When injected intravenously in mice, daintain/AIF1 at 75 pmol/kg inhibited glucose (1 g/kg)-stimulated insulin secretion, with a concomitant impairment of the glucose elimination, whereas at higher doses (7.5 and 75 nmol/kg), daintain/AIF1 potentiated glucose-stimulated insulin secretion and enhanced the glucose elimination. Its dual influence on insulin secretion in vivo at different peptide concentrations, and the abundance of macrophages expressing daintain/AIF1 in the pancreatic islets of prediabetic rats, suggest that daintain/AIF1 may have a role in connection with the pathogenesis of insulin-dependent diabetes mellitus. PMID- 9391122 TI - Tissue engineering of cartilage in space. AB - Tissue engineering of cartilage, i.e., the in vitro cultivation of cartilage cells on synthetic polymer scaffolds, was studied on the Mir Space Station and on Earth. Specifically, three-dimensional cell-polymer constructs consisting of bovine articular chondrocytes and polyglycolic acid scaffolds were grown in rotating bioreactors, first for 3 months on Earth and then for an additional 4 months on either Mir (10(-4)-10(-6) g) or Earth (1 g). This mission provided a unique opportunity to study the feasibility of long-term cell culture flight experiments and to assess the effects of spaceflight on the growth and function of a model musculoskeletal tissue. Both environments yielded cartilaginous constructs, each weighing between 0.3 and 0.4 g and consisting of viable, differentiated cells that synthesized proteoglycan and type II collagen. Compared with the Earth group, Mir-grown constructs were more spherical, smaller, and mechanically inferior. The same bioreactor system can be used for a variety of controlled microgravity studies of cartilage and other tissues. These results may have implications for human spaceflight, e.g., a Mars mission, and clinical medicine, e.g., improved understanding of the effects of pseudo-weightlessness in prolonged immobilization, hydrotherapy, and intrauterine development. PMID- 9391123 TI - Tissue-specific expression of herpes simplex virus thymidine kinase gene delivered by adeno-associated virus inhibits the growth of human hepatocellular carcinoma in athymic mice. AB - About 70% of hepatocellular carcinomas are known to express alpha-fetoprotein, which is normally expressed in fetal but not in adult livers. To induce herpes simplex virus-thymidine kinase expression in these cancer cells, we constructed an adeno-associated viral vector containing the HSV-TK gene under the control of the alpha-fetoprotein enhancer and albumin promoter. We previously demonstrated in vitro that although this vector can transduce a variety of human cells, only transduced AFP and albumin-expressing hepatocellular carcinoma cell lines were sensitive to killing by ganciclovir (GCV). In the present study, we explored the effect of this vector on hepatocellular carcinoma cells in vivo. Subcutaneous tumors generated in nude mice by implanting hepatocellular carcinoma cells previously transduced with this vector shrank dramatically after treatment with GCV. Bystander effect was also observed on the tumors generated by mixing transduced and untransduced cells. To test whether the tumor cells can be transduced by the virus in vivo, we injected the recombinant adeno-associated virus into tumors generated by untransduced hepatocarcinoma cell line. Tumor growth were retarded after treatment with GCV. These experiments demonstrate the feasibility of in vivo transduction of tumor cell with rAAV. PMID- 9391124 TI - Proliferation of adult T cell leukemia/lymphoma cells is associated with the constitutive activation of JAK/STAT proteins. AB - Human T cell leukemia/lymphotropic virus type I (HTLV-I) induces adult T cell leukemia/lymphoma (ATLL). The mechanism of HTLV-I oncogenesis in T cells remains partly elusive. In vitro, HTLV-I induces ligand-independent transformation of human CD4+ T cells, an event that correlates with acquisition of constitutive phosphorylation of Janus kinases (JAK) and signal transducers and activators of transcription (STAT) proteins. However, it is unclear whether the in vitro model of HTLV-I transformation has relevance to viral leukemogenesis in vivo. Here we tested the status of JAK/STAT phosphorylation and DNA-binding activity of STAT proteins in cell extracts of uncultured leukemic cells from 12 patients with ATLL by either DNA-binding assays, using DNA oligonucleotides specific for STAT-1 and STAT-3, STAT-5 and STAT-6 or, more directly, by immunoprecipitation and immunoblotting with anti-phosphotyrosine antibody for JAK and STAT proteins. Leukemic cells from 8 of 12 patients studied displayed constitutive DNA-binding activity of one or more STAT proteins, and the constitutive activation of the JAK/STAT pathway was found to persist over time in the 2 patients followed longitudinally. Furthermore, an association between JAK3 and STAT-1, STAT-3, and STAT-5 activation and cell-cycle progression was demonstrated by both propidium iodide staining and bromodeoxyuridine incorporation in cells of four patients tested. These results imply that JAK/STAT activation is associated with replication of leukemic cells and that therapeutic approaches aimed at JAK/STAT inhibition may be considered to halt neoplastic growth. PMID- 9391125 TI - Inflammatory mediators are perpetuated in macrophages resistant to apoptosis induced by hypoxia. AB - A hypoxic/anoxic microenvironment has been proposed to exist within a vascular lesion due to intimal or medial cell proliferation in vascular diseases. Here, we examined whether hypoxia alters macrophage function by exposing murine macrophage like RAW 264.7 (RAW) cells to hypoxia (2% O2). When cells were exposed to hypoxia, a significant number of RAW cells underwent apoptosis. Additionally, small subpopulations of RAW cells were resistant to hypoxia-induced apoptosis. Through repeated cycles of hypoxia exposure, hypoxia-induced apoptosis-resistant macrophages (HARMs) were selected; HARM cells demonstrate >70% resistance to hypoxia-induced apoptosis, as compared with the parental RAW cells. When heat shock protein (HSP) expression was examined after hypoxia, we observed a significant decrease in constitutive heat shock protein 70 (HSC 70) in RAW cells, but not in HARMs, as compared with the control normoxic condition (21% O2). In contrast, the expression level of glucose-regulated protein 78 (GRP 78) in RAW and HARM cells after hypoxia treatment was not altered, suggesting that HSC 70 and not GRP 78 may play a role in protection against hypoxia-induced apoptosis. When tumor necrosis factor alpha (TNF-alpha) production was examined after hypoxic treatment, a significant increase in TNF-alpha production in HARM but decrease in RAW was observed, as compared with cells cultured in normoxic conditions. HARM cells also exhibit a much lower level of modified-LDL uptake than do RAW cells, suggesting that HARMs may not transform into foam cells. These results suggest that a selective population of macrophages may adapt to potentially pathological hypoxic conditions by overcoming the apoptotic signal. PMID- 9391126 TI - How Escherichia coli can bias the results of molecular cloning: preferential selection of defective genomes of hepatitis C virus during the cloning procedure. AB - Cloned PCR products containing hepatitis C virus (HCV) genomic fragments have been used for analyses of HCV genomic heterogeneity and protein expression. These studies assume that the clones derived are representative of the entire virus population and that subsets are not inadvertently selected. The aim of the present study was to express HCV structural proteins. However, we found that there was a strong cloning selection for defective genomes and that most clones generated initially were incapable of expressing the HCV proteins. The HCV structural region (C-E1-E2-p7) was directly amplified by long reverse transcription-PCR from the plasma of an HCV-infected patient or from a control plasmid containing a viable full-length cDNA of HCV derived from the same patient but cloned in a different vector. The PCR products were cloned into a mammalian expression vector, amplified in Escherichia coli, and tested for their ability to produce HCV structural proteins. Twenty randomly picked clones derived from the HCV-infected patient all contained nucleotide mutations leading to absence or truncation of the expected HCV products. Of 25 clones derived from the control plasmid, only 8% were fully functional for polyprotein synthesis. The insertion of extra nucleotides in the region just upstream of the start codon of the HCV insert led to a statistically significant increase in the number of fully functional clones derived from the patient (42%) and from the control plasmid (72 92%). Nonrandom selection of clones during the cloning procedure has enormous implications for the study of viral heterogeneity, because it can produce a false spectrum of genomic diversity. It can also be an impediment to the construction of infectious viral clones. PMID- 9391127 TI - Tobacco smoke is a source of toxic reactive glycation products. AB - Smokers have a significantly higher risk for developing coronary and cerebrovascular disease than nonsmokers. Advanced glycation end products (AGEs) are reactive, cross-linking moieties that form from the reaction of reducing sugars and the amino groups of proteins, lipids, and nucleic acids. AGEs circulate in high concentrations in the plasma of patients with diabetes or renal insufficiency and have been linked to the accelerated vasculopathy seen in patients with these diseases. Because the curing of tobacco takes place under conditions that could lead to the formation of glycation products, we examined whether tobacco and tobacco smoke could generate these reactive species that would increase AGE formation in vivo. Our findings show that reactive glycation products are present in aqueous extracts of tobacco and in tobacco smoke in a form that can rapidly react with proteins to form AGEs. This reaction can be inhibited by aminoguanidine, a known inhibitor of AGE formation. We have named these glycation products "glycotoxins." Like other known reducing sugars and reactive glycation products, glycotoxins form smoke, react with protein, exhibit a specific fluorescence when cross-linked to proteins, and are mutagenic. Glycotoxins are transferred to the serum proteins of human smokers. AGE apolipoprotein B and serum AGE levels in cigarette smokers were significantly higher than those in nonsmokers. These results suggest that increased glycotoxin exposure may contribute to the increased incidence of atherosclerosis and high prevalence of cancer in smokers. PMID- 9391128 TI - Long-term correction of obesity and diabetes in genetically obese mice by a single intramuscular injection of recombinant adeno-associated virus encoding mouse leptin. AB - The ob/ob mouse is genetically deficient in leptin and exhibits a phenotype that includes obesity and non-insulin-dependent diabetes mellitus. This phenotype closely resembles the morbid obesity seen in humans. In this study, we demonstrate that a single intramuscular injection of a recombinant adeno associated virus (AAV) vector encoding mouse leptin (rAAV-leptin) in ob/ob mice leads to prevention of obesity and diabetes. The treated animals show normalization of metabolic abnormalities including hyperglycemia, insulin resistance, impaired glucose tolerance, and lethargy. The effects of a single injection have lasted through the 6-month course of the study. At all time points measured the circulating levels of leptin in the serum were similar to age matched control C57 mice. These results demonstrate that maintenance of normal levels of leptin (2-5 ng/ml) in the circulation can prevent both the onset of obesity and associated non-insulin-dependent diabetes. Thus a single injection of a rAAV vector expressing a therapeutic gene can lead to complete and long-term correction of a genetic disorder. Our study demonstrates the long-term correction of a disease caused by a genetic defect and proves the feasibility of using rAAV based vectors for the treatment of chronic disorders like obesity. PMID- 9391129 TI - Polyclonal structure of intestinal adenomas in ApcMin/+ mice with concomitant loss of Apc+ from all tumor lineages. AB - When tumors form in intestinal epithelia, it is important to know whether they involve single initiated somatic clones. Advanced carcinomas in humans and mice are known to be monoclonal. However, earlier stages of tumorigenesis may instead involve an interaction between cells that belong to separate somatic clones within the epithelium. The clonality of early tumors has been investigated in mice with an inherited predisposition to intestinal tumors. Analysis of Min (multiple intestinal neoplasia) mice chimeric for a ubiquitously expressed cell lineage marker revealed that normal intestinal crypts are monoclonal, but intestinal adenomas frequently have a polyclonal structure, presenting even when very small as single, focal adenomas composed of at least two somatic lineages. Furthermore, within these polyclonal adenomas, all tumor lineages frequently lose the wild-type Apc allele. These observations can be interpreted by several models for clonal interaction within the epithelium, ranging from passive fusion within regions of high neoplastic potential to a requirement for active clonal cooperation. PMID- 9391130 TI - Interactions of the chaperone Hsp104 with yeast Sup35 and mammalian PrP. AB - [PSI+] is a genetic element in yeast for which a heritable change in phenotype appears to be caused by a heritable change in the conformational state of the Sup35 protein. The inheritance of [PSI+] and the physical state of Sup35 in vivo depend on the protein chaperone Hsp104 (heat shock protein 104). Although these observations provide a strong genetic argument in support of the "protein-only" or "prion" hypothesis for [PSI+], there is, as yet, no direct evidence of an interaction between the two proteins. We report that when purified Sup35 and Hsp104 are mixed, the circular dichroism (CD) spectrum differs from that predicted by the addition of the proteins' individual spectra, and the ATPase activity of Hsp104 is inhibited. Similar results are obtained with two other amyloidogenic substrates, mammalian PrP and beta-amyloid 1-42 peptide, but not with several control proteins. With a group of peptides that span the PrP protein sequence, those that produced the largest changes in CD spectra also caused the strongest inhibition of ATPase activity in Hsp104. Our observations suggest that (i) previously described genetic interactions between Hsp104 and [PSI+] are caused by direct interaction between Hsp104 and Sup35; (ii) Sup35 and PrP, the determinants of the yeast and mammalian prions, respectively, share structural features that lead to a specific interaction with Hsp104; and (iii) these interactions couple a change in structure to the ATPase activity of Hsp104. PMID- 9391131 TI - Chaperone-supervised conversion of prion protein to its protease-resistant form. AB - Transmissible spongiform encephalopathies (TSEs) are lethal, infectious disorders of the mammalian nervous system. A TSE hallmark is the conversion of the cellular protein PrPC to disease-associated PrPSc (named for scrapie, the first known TSE). PrPC is protease-sensitive, monomeric, detergent soluble, and primarily alpha-helical; PrPSc is protease-resistant, polymerized, detergent insoluble, and rich in beta-sheet. The "protein-only" hypothesis posits that PrPSc is the infectious TSE agent that directly converts host-encoded PrPC to fresh PrPSc, harming neurons and creating new agents of infection. To gain insight on the conformational transitions of PrP, we tested the ability of several protein chaperones, which supervise the conformational transitions of proteins in diverse ways, to affect conversion of PrPC to its protease-resistant state. None affected conversion in the absence of pre-existing PrPSc. In its presence, only two, GroEL and Hsp104 (heat shock protein 104), significantly affected conversion. Both promoted it, but the reaction characteristics of conversions with the two chaperones were distinct. In contrast, chemical chaperones inhibited conversion. Our findings provide new mechanistic insights into nature of PrP conversions, and provide a new set of tools for studying the process underlying TSE pathogenesis. PMID- 9391132 TI - Mutations in dihydropteroate synthase are responsible for sulfone and sulfonamide resistance in Plasmodium falciparum. AB - Plasmodium falciparum causes the most severe form of malaria in humans. An important class of drugs in malaria treatment is the sulfone/sulfonamide group, of which sulfadoxine is the most commonly used. The target of sulfadoxine is the enzyme dihydropteroate synthase (DHPS), and sequencing of the DHPS gene has identified amino acid differences that may be involved in the mechanism of resistance to this drug. In this study we have sequenced the DHPS gene in 10 isolates from Thailand and identified a new allele of DHPS that has a previously unidentified amino acid difference. We have expressed eight alleles of P. falciparum PPPK-DHPS in Escherichia coli and purified the functional enzymes to homogeneity. Strikingly, the Ki for sulfadoxine varies by almost three orders of magnitude from 0.14 microM for the DHPS allele from sensitive isolates to 112 microM for an enzyme expressed in a highly resistant isolate. Comparison of the Ki of different sulfonamides and the sulfone dapsone has suggested that the amino acid differences in DHPS would confer cross-resistance to these compounds. These results show that the amino acid differences in the DHPS enzyme of sulfadoxine resistant isolates of P. falciparum are central to the mechanism of resistance to sulfones and sulfonamides. PMID- 9391133 TI - Backtracking leukemia to birth: identification of clonotypic gene fusion sequences in neonatal blood spots. AB - Epidemiological evidence has suggested that some pediatric leukemias may be initiated in utero and, for some pairs of identical twins with concordant leukemia, this possibility has been strongly endorsed by molecular studies of clonality. Direct evidence for a prenatal origin can only be derived by prospective or retrospective detection of leukemia-specific molecular abnormalities in fetal or newborn samples. We report a PCR-based method that has been developed to scrutinize neonatal blood spots (Guthrie cards) for the presence of numerically infrequent leukemic cells at birth in individuals who subsequently developed leukemia. We demonstrate that unique or clonotypic MLL-AF4 genomic fusion sequences are present and detectable in neonatal blood spots from individuals who were diagnosed with acute lymphoblastic leukemia at ages 5 months to 2 years and, therefore, have arisen during fetal hematopoiesis in utero. This result provides unequivocal evidence for a prenatal initiation of acute leukemia in young patients. The method should be applicable to other fusion genes in children with common subtypes of leukemia and will be of value in attempts to unravel the natural history and etiology of this major subtype of pediatric cancer. PMID- 9391134 TI - A dichotomous role for nitric oxide during acute Toxoplasma gondii infection in mice. AB - Production of nitric oxide by macrophages is believed to be an important microbicidal mechanism for a variety of intracellular pathogens, including Toxoplasma gondii. Mice with a targeted disruption of the inducible nitric oxide synthase gene (iNOS) were infected orally with T. gondii tissue cysts. Time to death was prolonged compared with parental controls. Histologic analysis of tissue from infected mice showed scattered small foci of inflammation with parasites in various tissues of iNOS-/- mice, whereas tissue from the parental C57BL/6 mice had more extensive tissue inflammation with few visible parasites. In particular, extensive ulceration and necrosis of distal small intestine and fatty degeneration of the liver was seen in the parental mice at day 7 postinfection, as compared with the iNOS-/- mice where these tissues appeared normal. Serum interferon gamma and tumor necrosis factor alpha levels postinfection were equally elevated in both mouse strains. Treatment of the parental mice with a NO synthase inhibitor, aminoguanidine, prevented early death in these mice as well as the hepatic degeneration and small bowel necrosis seen in acutely infected control parentals. These findings indicate that NO production during acute infection with T. gondii can kill intracellular parasites but can be detrimental, even lethal, to the host. PMID- 9391135 TI - Respiratory syncytial virus (RSV) SH and G proteins are not essential for viral replication in vitro: clinical evaluation and molecular characterization of a cold-passaged, attenuated RSV subgroup B mutant. AB - A live, cold-passaged (cp) candidate vaccine virus, designated respiratory syncytial virus (RSV) B1 cp-52/2B5 (cp-52), replicated efficiently in Vero cells, but was found to be overattenuated for RSV-seronegative infants and children. Sequence analysis of reverse-transcription-PCR-amplified fragments of this mutant revealed a large deletion spanning most of the coding sequences for the small hydrophobic (SH) and attachment (G) proteins. Northern blot analysis of cp-52 detected multiple unique read-through mRNAs containing SH and G sequences, consistent with a deletion mutation spanning the SH:G gene junction. Immunological studies confirmed that an intact G glycoprotein was not produced by the cp-52 virus. Nonetheless, cp-52 was infectious and replicated to high titer in tissue culture despite the absence of the viral surface SH and G glycoproteins. Thus, our characterization of this negative-strand RNA virus identified a novel replication-competent deletion mutant lacking two of its three surface glycoproteins. The requirement of SH and G for efficient replication in vivo suggests that selective deletion of one or both of these RSV genes may provide an alternative or additive strategy for developing an optimally attenuated vaccine candidate. PMID- 9391136 TI - Two forms of replication initiator protein: positive and negative controls. AB - The pir gene of plasmid R6K encodes the protein, pi, a replication and transcription factor. Two translational options for the pir gene give rise to two forms of pi protein: a 35.0-kDa form (pi35.0) and a shortened 30.5-kDa form (pi30.5). Although both proteins bind to a series of 22-bp direct repeats essential for plasmid R6K replication, only pi35.0 can bind to a site in the (A.T)-rich segment of its gamma ori and activate the gamma ori in vivo and in vitro. However, unlike pi35.0, pi30.5can inhibit in vivo and in vitro replication (activated by pi35.0). We propose that the two forms of pi might have distinct functions in replication. We show that although both forms of pi produce dimers, the nature of these dimers is not identical. The N-terminal 37 amino acid residues appear to control the formation of the more stable pi35.0 dimers, whereas another, apparently weaker interface holds together dimers of pi30.5. We speculate that the leucine zipper-like motif, absent in pi30.5, controls very specific functions of pi protein. PMID- 9391137 TI - Us9, a stable lysine-less herpes simplex virus 1 protein, is ubiquitinated before packaging into virions and associates with proteasomes. AB - The US9 gene of herpes simplex virus 1 encodes a virion tegument protein with a predicted Mr of 10,000. Earlier studies have shown that the gene is not essential for viral replication in cells in culture. We report that (i) US9 forms in denaturing polyacrylamide gels multiple overlapping bands ranging in Mr from 12,000 to 25,000; (ii) the protein recovered from infected cells or purified virions reacts with anti-ubiquitin antibodies; (iii) autoradiographic images of US9 protein immunoprecipitated from cells infected with [35S]methionine-labeled virus indicate that the protein is stable for at least 4 h after entry into cells (the protein was also stable for at least 4 h after a 1-h labeling interval 12 h after infection); (iv) antibody to subunit 12 of proteasomes pulls down US9 protein from herpes simplex virus-infected cell lysates; and (v) the US9 gene is highly conserved among the members of the alpha subfamily of herpes viruses, and the US9 gene product lacks lysines. We conclude that US9 is a lysine-less, ubiquitinated protein that interacts with the ubiquitin-dependent pathway for degradation of proteins and that this function may be initiated at the time of entry of the virus into the cell. PMID- 9391138 TI - Loss of HMW1 and HMW3 in noncytadhering mutants of Mycoplasma pneumoniae occurs post-translationally. AB - The genomic sequence of Mycoplasma pneumoniae establish this cell-wall-less prokaryote as among the smallest known microorganisms capable of self replication. However, this genomic simplicity and corresponding biosynthetic austerity are sharply contrasted by the complex terminal structure found in this species. This tip structure (attachment organelle) directs colonization of the human respiratory mucosa, leading to bronchitis and atypical pneumonia. Furthermore, formation of a second tip structure appears to precede cell division, implying temporal regulation. However, the organization, regulation, and assembly of the attachment organelle in M. pneumoniae are poorly understood, and no counterparts have been identified among the walled bacteria. M. pneumoniae possesses a cytoskeleton-like structure required to localize adhesin proteins to the attachment organelle. The cytadherence-associated proteins HMW1, HMW2, and HMW3 are components of the mycoplasma cytoskeleton, with HMW1 localizing strictly along the filamentous extensions from the cell body and HMW3 being a key structural element of the terminal organelle. Disruptions in hmw2 result in the loss of HMW1 and HMW3. However, the hmw1 and hmw3 genes were transcribed and translated at wild-type levels in hmw2 mutants. HMW1 and HMW3 were relatively stable in the wild-type background over 8 h but disappeared in the mutants over this time period. Evaluation of recombinant HMW1 levels in mycoplasma mutants suggested a requirement for the C-terminal domain of HMW1 for turnover. Finally, an apparent defect in the processing of the precursor for the adhesin protein P1 was noted in the HMW- mutants. PMID- 9391139 TI - Use of differential display analysis to assess the effect of human cytomegalovirus infection on the accumulation of cellular RNAs: induction of interferon-responsive RNAs. AB - We used differential display analysis to identify mRNAs that accumulate to enhanced levels in human cytomegalovirus-infected cells as compared with mock infected cells. RNAs were compared at 8 hr after infection of primary human fibroblasts. Fifty-seven partial cDNA clones were isolated, representing about 26 differentially expressed mRNAs. Eleven of the mRNAs were virus-coded, and 15 were of cellular origin. Six of the partial cDNA sequences have not been reported previously. All of the cellular mRNAs identified in the screen are induced by interferon alpha. The induction in virus-infected cells, however, does not involve the action of interferon or other small signaling molecules. Neutralizing antibodies that block virus infection also block the induction. These RNAs accumulate after infection with virus that has been inactivated by treatment with UV light, indicating that the inducer is present in virions. We conclude that human cytomegalovirus induces interferon-responsive mRNAs. PMID- 9391140 TI - DNA topoisomerase targets of the fluoroquinolones: a strategy for avoiding bacterial resistance. AB - Fluoroquinolones are antibacterial agents that attack DNA gyrase and topoisomerase IV on chromosomal DNA. The existence of two fluoroquinolone targets and stepwise accumulation of resistance suggested that new quinolones could be found that would require cells to obtain two topoisomerase mutations to display resistance. For wild-type cells to become resistant, the two mutations must be acquired concomitantly. That is expected to occur infrequently. To identify such compounds, fluoroquinolones were tested for the ability to kill a moderately resistant gyrase mutant. Compounds containing a C8-methoxyl group were particularly lethal, and incubation of wild-type cultures on agar containing C8 methoxyl fluoroquinolones produced no resistant mutant, whereas thousands arose during comparable treatment with control compounds lacking the C8 substituent. When the test strain contained a preexisting topoisomerase IV mutation, which by itself conferred no resistance, equally high numbers of resistant mutants were obtained for C8-methoxyl and control compounds. Thus C8-methoxyl fluoroquinolones required two mutations for expression of resistance. Although highly lethal, C8 methoxyl fluoroquinolones were not more effective than C8-H controls at blocking bacterial growth. Consequently, quinolone action involves two events, which we envision as formation of drug-enzyme-DNA complexes followed by release of lethal double-strand DNA breaks. Release of DNA breaks, which must occur less frequently than complex formation, is probably the process stimulated by the C8-methoxyl group. Understanding this stimulation should provide insight into intracellular quinolone action and contribute to development of fluoroquinolones that prevent selection of resistant bacteria. PMID- 9391141 TI - Periplasmic superoxide dismutase protects Salmonella from products of phagocyte NADPH-oxidase and nitric oxide synthase. AB - Superoxide dismutase (SOD) catalyzes the conversion of superoxide radical to hydrogen peroxide. Periplasmic localization of bacterial Cu,Zn-SOD has suggested a role of this enzyme in defense against extracellular phagocyte-derived reactive oxygen species. Sequence analysis of regions flanking the Salmonella typhimurium sodC gene encoding Cu,Zn-SOD demonstrates significant homology to lambda phage proteins, reflecting possible bacteriophage-mediated horizontal gene transfer of this determinant among pathogenic bacteria. Salmonella deficient in Cu,Zn-SOD has reduced survival in macrophages and attenuated virulence in mice, which can be restored by abrogation of either the phagocyte respiratory burst or inducible nitric oxide synthase. Moreover, a sodC mutant is extremely susceptible to the combination of superoxide and nitric oxide. These observations suggest that SOD protects periplasmic or inner membrane targets by diverting superoxide and limiting peroxynitrite formation, and they demonstrate the ability of the respiratory burst and nitric oxide synthase to synergistically kill microbial pathogens in vivo. PMID- 9391142 TI - Chronic stress alters synaptic terminal structure in hippocampus. AB - Repeated psychosocial or restraint stress causes atrophy of apical dendrites in CA3 pyramidal neurons of the hippocampus, accompanied by specific cognitive deficits in spatial learning and memory. Excitatory amino acids mediate this atrophy together with adrenal steroids and the neurotransmitter serotonin. Because the mossy fibers from dentate granule neurons provide a major excitatory input to the CA3 proximal apical dendrites, we measured ultrastructural parameters associated with the mossy fiber-CA3 synapses in control and 21-day restraint-stressed rats in an effort to find additional morphological consequences of stress that could help elucidate the underlying anatomical as well as cellular and molecular mechanisms. Although mossy fiber terminals of control rats were packed with small, clear synaptic vesicles, terminals from stressed animals showed a marked rearrangement of vesicles, with more densely packed clusters localized in the vicinity of active zones. Moreover, compared with controls, restraint stress increased the area of the mossy fiber terminal occupied by mitochondrial profiles and consequently, a larger, localized energy generating capacity. A single stress session did not produce these changes either immediately after or the next day following the restraint session. These findings provide a morphological marker of the effects of chronic stress on the hippocampus that points to possible underlying neuroanatomical as well as cellular and molecular mechanisms for the ability of repeated stress to cause structural changes within the hippocampus. PMID- 9391143 TI - Induction of long-term depression and rebound potentiation by inositol trisphosphate in cerebellar Purkinje neurons. AB - Cerebellar Purkinje neurons receive two major excitatory inputs, the climbing fibers (CFs) and parallel fibers (PFs). Simultaneous, repeated activation of CFs and PFs results in the long-term depression (LTD) of the amplitude of PF-evoked synaptic currents. To induce LTD, activation of CFs may be substituted with depolarization of the Purkinje neuron to turn on voltage-activated calcium channels and increase the intracellular calcium concentration. The role of PFs in the induction of LTD, however, is less clear. PFs activate glutamate metabotropic receptors that increase phosphoinositide turnover and elevate cytosolic inositol 1,4,5-trisphosphate (InsP3). It has been proposed that calcium release from intracellular stores via InsP3 receptors may be important in the induction of LTD. We studied the role of InsP3 in the induction of LTD by photolytic release of InsP3 from its biologically inactive "caged" precursor in voltage-clamped Purkinje neurons in acutely prepared cerebellar slices. We find that InsP3-evoked calcium release is as effective in LTD induction as activation of PFs. InsP3 induced LTD was prevented by calcium chelator 1,2-bis(2-amino phenoxy)ethane N,N,N', N'-tetraacetic acid. LTD produced either by repeated activation of PFs combined with depolarization (PF+DeltaV), or by InsP3 combined with depolarization (InsP3+DeltaV) saturated at approximately 50%. Maximal LTD induced by PF+DeltaV could not be further increased by InsP3+DeltaV and vice versa, which suggests that both protocols for induction of LTD share a common path. In addition to inducing LTD, photo-release of InsP3+DeltaV resulted in the rebound potentiation of inhibitory synaptic currents. In the presence of heparin, an InsP3 receptor antagonist, repeated activation of PF+DeltaV failed to induce LTD, suggesting that InsP3 receptors play an important role in LTD induction under physiological conditions. PMID- 9391144 TI - Functional activation in motor cortex reflects the direction and the degree of handedness. AB - Handedness is the clearest example of behavioral lateralization in humans. It is not known whether the obvious asymmetry manifested by hand preference is associated with similar asymmetry in brain activation during movement. We examined the functional activation in cortical motor areas during movement of the dominant and nondominant hand in groups of right-handed and left-handed subjects and found that use of the dominant hand was associated with a greater volume of activation in the contralateral motor cortex. Furthermore, there was a separate relation between the degree of handedness and the extent of functional lateralization in the motor cortex. The patterns of functional activation associated with the direction and degree of handedness suggest that these aspects are independent and are coded separately in the brain. PMID- 9391145 TI - Syntax processing by auditory cortical neurons in the FM-FM area of the mustached bat Pteronotus parnellii. AB - Syntax denotes a rule system that allows one to predict the sequencing of communication signals. Despite its significance for both human speech processing and animal acoustic communication, the representation of syntactic structure in the mammalian brain has not been studied electrophysiologically at the single unit level. In the search for a neuronal correlate for syntax, we used playback of natural and temporally destructured complex species-specific communication calls-so-called composites-while recording extracellularly from neurons in a physiologically well defined area (the FM-FM area) of the mustached bat's auditory cortex. Even though this area is known to be involved in the processing of target distance information for echolocation, we found that units in the FM-FM area were highly responsive to composites. The finding that neuronal responses were strongly affected by manipulation in the time domain of the natural composite structure lends support to the hypothesis that syntax processing in mammals occurs at least at the level of the nonprimary auditory cortex. PMID- 9391146 TI - The seven-transmembrane spanning topography of the Alzheimer disease-related presenilin proteins in the plasma membranes of cultured cells. AB - To ascertain the membrane topography of the multi-transmembrane spanning presenilin proteins PS-1 and PS-2, anti-peptide antibodies were raised to several specific amino acid sequences in the two proteins, and, after their specificity was ascertained, the anti-peptide antibodies were used in immunofluorescent labeling of live PS-transfected, cultured DAMI cells, which are impermeable to the antibodies, as well as of their fixed and permeabilized counterparts. In such experiments, antibodies that specifically stain the intact live cells must label epitopes of the PS proteins that are on the exterior face of the plasma membrane whereas those antibodies that do not stain the live cells but do stain the fixed and permeabilized cells must label epitopes that face the cytoplasmic side of the membrane. The results obtained were entirely in accord with the predictions of the seven-transmembrane spanning topography (like that of rhodopsin and the beta adrenergic receptor) and were totally inconsistent with the expectations for either the six- or eight-transmembrane topographies that have been proposed. PMID- 9391147 TI - On the spurious endoproteolytic processing of the presenilin proteins in cultured cells and tissues. AB - It has been widely reported that the presenilin proteins PS-1 and PS-2 in extracts derived from a variety of cultured cells and from tissues are fragmented extensively by endoproteolytic processing events. It generally has been presumed that this endoproteolysis is a physiologically normal intracellular event following presenilin expression, which might play an important role in the still unknown functions of these molecules in connection with Alzheimer disease. We demonstrate herein, however, that, if a variety of cultured cells and several mouse tissues are examined under conditions minimizing cell trauma, the presenilin molecules in the extracts are found to be intact but that, if the cells and tissues are prepared under somewhat more stressful conditions, the endoproteolytic fragments are then observed. We conclude that these particular endoproteolytic events are not the result of physiologically normal processing of the presenilins but are rather artifacts occurring during the common procedures of specimen preparation. PMID- 9391148 TI - Widespread expression and estrogen regulation of PPEIA-3' nuclear RNA in the rat brain. AB - We previously identified a novel nuclear RNA species derived from the preproenkephalin (PPE) gene. This transcript, which we have named PPEIA-3' RNA, hybridizes with probes directed at a region of PPE intron A downstream of an alternative germ-cell transcription start site, but does not contain PPE protein coding sequences. We now report that estrogen treatment of ovariectomized rats increases the expression of conventional PPE heteronuclear RNA, and also induces the expression of PPEIA-3' RNA, apparently in separate cell populations within the ventromedial nucleus of the hypothalamus. Further, we show that cells expressing PPEIA-3' are found in several neuronal groups in the rat forebrain and brainstem, with a distinct topographical distribution. High densities of PPEIA-3' containing cells are found in the reticular thalamic nucleus, the basal forebrain, the vestibular complex, the deep cerebellar nuclei, and the trapezoid body, a pattern that parallels the distribution of atypical nuclear RNAs described by other groups. These results suggest that this diverse neuronal population shares a common set of nuclear factors responsible for the expression and retention of this atypical RNA transcript. The implication of these results for cell-specific gene transcription and regulation in the brain and the possible relationship of PPEIA-3' RNA and other atypical nuclear RNAs is discussed. PMID- 9391149 TI - beta subunits influence the biophysical and pharmacological differences between P and Q-type calcium currents expressed in a mammalian cell line. AB - Human epithelial kidney cells (HEK) were prepared to coexpress alpha1A, alpha2delta with different beta calcium channel subunits and green fluorescence protein. To compare the calcium currents observed in these cells with the native neuronal currents, electrophysiological and pharmacological tools were used conjointly. Whole-cell current recordings of human epithelial kidney alpha1A transfected cells showed small inactivating currents in 80 mM Ba2+ that were relatively insensitive to calcium blockers. Coexpression of alpha1A, betaIb, and alpha2delta produced a robust inactivating current detected in 10 mM Ba2+, reversibly blockable with low concentration of omega-agatoxin IVA (omega-Aga IVA) or synthetic funnel-web spider toxin (sFTX). Barium currents were also supported by alpha1A, beta2a, alpha2delta subunits, which demonstrated the slowest inactivation and were relatively insensitive to omega-Aga IVA and sFTX. Coexpression of beta3 with the same combination as above produced inactivating currents also insensitive to low concentration of omega-Aga IVA and sFTX. These data indicate that the combination alpha1A, betaIb, alpha2delta best resembles P type channels given the rate of inactivation and the high sensitivity to omega Aga IVA and sFTX. More importantly, the specificity of the channel blocker is highly influenced by the beta subunit associated with the alpha1A subunit. PMID- 9391150 TI - Glucocorticoid enhancement of memory storage involves noradrenergic activation in the basolateral amygdala. AB - Evidence indicates that the modulatory effects of the adrenergic stress hormone epinephrine as well as several other neuromodulatory systems on memory storage are mediated by activation of beta-adrenergic mechanisms in the amygdala. In view of our recent findings indicating that the amygdala is involved in mediating the effects of glucocorticoids on memory storage, the present study examined whether the glucocorticoid-induced effects on memory storage depend on beta-adrenergic activation within the amygdala. Microinfusions (0.5 microg in 0.2 microl) of either propranolol (a nonspecific beta-adrenergic antagonist), atenolol (a beta1 adrenergic antagonist), or zinterol (a beta2-adrenergic antagonist) administered bilaterally into the basolateral nucleus of the amygdala (BLA) of male Sprague Dawley rats 10 min before training blocked the enhancing effect of posttraining systemic injections of dexamethasone (0.3 mg/kg) on 48-h memory for inhibitory avoidance training. Infusions of these beta-adrenergic antagonists into the central nucleus of the amygdala did not block the dexamethasone-induced memory enhancement. Furthermore, atenolol (0.5 microg) blocked the memory-enhancing effects of the specific glucocorticoid receptor (GR or type II) agonist RU 28362 infused concurrently into the BLA immediately posttraining. These results strongly suggest that beta-adrenergic activation is an essential step in mediating glucocorticoid effects on memory storage and that the BLA is a locus of interaction for these two systems. PMID- 9391151 TI - Insulin-like growth factors-I and -II differentially regulate endogenous acetylcholine release from the rat hippocampal formation. AB - Insulin-like growth factors-I and -II (IGF-I and -II) are structurally related mitogenic polypeptides with potent growth promoting effects. These peptides and their corresponding IGF-I and -II receptors are selectively localized in the brain. To date, most of the effects of IGFs are believed to be mediated by IGF-I receptors whereas the significance of IGF-II receptor in mediating biological responses remains unclear. In the present study, we characterized the distribution of IGF-I and IGF-II receptor sites and investigated the effects of both factors on endogenous acetylcholine (ACh) release in adult rat hippocampus. [125I]IGF-I receptor binding sites are recognized by IGF-I> IGF-II> insulin, whereas [125I]IGF-II binding was competed potently by IGF-II> IGF-I but not by insulin. At the cellular level, IGF-I receptor sites were primarily noted in the molecular layer of the dentate gyrus and the CA2-CA3 subfields of the Ammon's horn whereas IGF-II sites were localized predominantly in the pyramidal cell layer of the CA1-CA3 subfields and in the granular cell layer of the dentate gyrus. IGF-I (10(-14)-10(-8) M) and des(1-3) IGF-I (10(-10)-10(-8) M) were found to inhibit whereas IGF-II (10(-14)-10(-8) M) potentiated K+-evoked ACh release from hippocampal slices. Tetrodotoxin altered the effects of IGF-I but not those of IGF-II suggesting that IGF-I acts indirectly via the release of other modulators whereas IGF-II acts directly on or in close proximity to the cholinergic terminals. The inhibitory effects of IGF-I were also observed in the frontal cortex but not in the striatum. In contrast, the stimulatory effects of IGF-II were evident both in the frontal cortex and striatum. Taken together, these results reveal the differential localization of IGF-I and IGF-II receptor sites in the hippocampal formation and the opposite role for these growth factors in the acute regulation of ACh release likely via two distinct mechanisms. Additionally, these data provide the first evidence for a direct role for IGF-II and its receptors in the regulation of transmitter release in the central nervous system. PMID- 9391152 TI - Epilepsy in mice deficient in the 65-kDa isoform of glutamic acid decarboxylase. AB - gamma-Aminobutyric acid (GABA), the major inhibitory neurotransmitter in the mammalian brain, is synthesized by two glutamate decarboxylase isoforms, GAD65 and GAD67. The separate role of the two isoforms is unknown, but differences in saturation with cofactor and subcellular localization suggest that GAD65 may provide reserve pools of GABA for regulation of inhibitory neurotransmission. We have disrupted the gene encoding GAD65 and backcrossed the mutation into the C57BL/6 strain of mice. In contrast to GAD67-/- animals, which are born with developmental abnormalities and die shortly after birth, GAD65-/- mice appear normal at birth. Basal GABA levels and holo-GAD activity are normal, but the pyridoxal 5' phosphate-inducible apo-enzyme reservoir is significantly decreased. GAD65-/- mice develop spontaneous seizures that result in increased mortality. Seizures can be precipitated by fear or mild stress. Seizure susceptibility is dramatically increased in GAD65-/- mice backcrossed into a second genetic background, the nonobese diabetic (NOD/LtJ) strain of mice enabling electroencephalogram analysis of the seizures. The generally higher basal brain GABA levels in this backcross are significantly decreased by the GAD65-/- mutation, suggesting that the relative contribution of GABA synthesized by GAD65 to total brain GABA levels is genetically determined. Seizure-associated c-fos like immunoreactivity reveals the involvement of limbic regions of the brain. These data suggest that GABA synthesized by GAD65 is important in the dynamic regulation of neural network excitability, implicate at least one modifier locus in the NOD/LtJ strain, and present GAD65-/- animals as a model of epilepsy involving GABA-ergic pathways. PMID- 9391153 TI - Large conductance voltage- and calcium-dependent K+ channel, a distinct member of voltage-dependent ion channels with seven N-terminal transmembrane segments (S0 S6), an extracellular N terminus, and an intracellular (S9-S10) C terminus. AB - Large conductance voltage- and Ca2+-dependent K+ (MaxiK) channels show sequence similarities to voltage-gated ion channels. They have a homologous S1-S6 region, but are unique at the N and C termini. At the C terminus, MaxiK channels have four additional hydrophobic regions (S7-S10) of unknown topology. At the N terminus, we have recently proposed a new model where MaxiK channels have an additional transmembrane region (S0) that confers beta subunit regulation. Using transient expression of epitope tagged MaxiK channels, in vitro translation, functional, and "in vivo" reconstitution assays, we now show that MaxiK channels have seven transmembrane segments (S0-S6) at the N terminus and a S1-S6 region that folds in a similar way as in voltage-gated ion channels. Further, our results indicate that hydrophobic segments S9-S10 in the C terminus are cytoplasmic and unequivocally demonstrate that S0 forms an additional transmembrane segment leading to an exoplasmic N terminus. PMID- 9391154 TI - Altered gene expression in the host brain caused by a trematode parasite: neuropeptide genes are preferentially affected during parasitosis. AB - Schistosome parasites adjust the physiology and behavior of their intermediate molluscan hosts to their own benefit. Previous studies demonstrated effects of the avian-schistosome Trichobilharzia ocellata on peptidergic centers in the brain of the intermediate snail host Lymnaea stagnalis. In particular, electrophysiological properties and peptide release of growth- and reproduction controlling neuroendocrine neurons were affected. We now have examined the possibility that the expression of genes that control physiology and behavior of the host might be altered during parasitosis. A cDNA library of the brain of parasitized Lymnaea was constructed and differentially screened by using mRNA from the brain of both parasitized and nonparasitized snails. This screening yielded a number of clones, including previously identified cDNAs as well as novel neuronal transcripts, which appear to be differentially regulated. The majority of these transcripts encode neuropeptides. Reverse Northern blot analysis confirmed that neuropeptide gene expression is indeed affected in parasitized animals. Moreover, the expression profiles of 10 transcripts tested showed a differential, parasitic stage-specific regulation. Changes in expression could in many cases already be observed between 1.5 and 5 hr postinfection, suggesting that changes in gene expression are a direct effect of parasitosis. We suggest that direct regulation of neuropeptide gene expression is a strategy of parasites to induce physiological and behavioral changes in the host. PMID- 9391155 TI - Estrogen receptor-dependent sexual differentiation of dopaminergic neurons in the preoptic region of the mouse. AB - Although it has been known for some time that estrogen exerts a profound influence on brain development a definitive demonstration of the role of the classical estrogen receptor (ERalpha) in sexual differentiation has remained elusive. In the present study we used a sexually dimorphic population of dopaminergic neurons in the anteroventral periventricular nucleus of the hypothalamus (AVPV) to test the dependence of sexual differentiation on a functional ERalpha by comparing the number of tyrosine hydroxylase (TH) immunoreactive neurons in the AVPV of wild-type (WT) mice with that of mice in which the ERalpha had been disrupted by homologous recombination (ERKOalpha). Only a few ERalpha-immunoreactive neurons were detected in the AVPV of ERKOalpha mice, and the number of TH-immunoreactive neurons was three times that of WT mice, suggesting that disruption of the ERalpha gene feminized the number of TH immunoreactive neurons. In contrast, the AVPV contains the same number of TH immunoreactive neurons in testicular feminized male mice as in WT males, indicating that sexual differentiation of this population of neurons is not dependent on an intact androgen receptor. The number of TH-immunoreactive neurons in the AVPV of female ERKOalpha mice remained higher than that of WT males, but TH staining appeared to be lower than that of WT females. Thus, the sexual differentiation of dopamine neurons in the AVPV appears to be receptor specific and dependent on the perinatal steroid environment. PMID- 9391156 TI - Midbrain injection of recombinant adeno-associated virus encoding rat glial cell line-derived neurotrophic factor protects nigral neurons in a progressive 6 hydroxydopamine-induced degeneration model of Parkinson's disease in rats. AB - A recombinant adeno-associated virus (rAAV) vector capable of infecting cells and expressing rat glial cell line-derived neurotrophic factor (rGDNF), a putative central nervous system dopaminergic survival factor, under the control of a potent cytomegalovirus (CMV) immediate/early promoter (AAV-MD-rGDNF) was constructed. Two experiments were performed to evaluate the time course of expression of rAAV-mediated GDNF protein expression and to test the vector in an animal model of Parkinson's disease. To evaluate the ability of rAAV-rGDNF to protect nigral dopaminergic neurons in the progressive Sauer and Oertel 6 hydroxydopamine (6-OHDA) lesion model, rats received perinigral injections of either rAAV-rGDNF virus or rAAV-lacZ control virus 3 weeks prior to a striatal 6 OHDA lesion and were sacrificed 4 weeks after 6-OHDA. Cell counts of back-labeled fluorogold-positive neurons in the substantia nigra revealed that rAAV-MD-rGDNF protected a significant number of cells when compared with cell counts of rAAV CMV-lacZ-injected rats (94% vs. 51%, respectively). In close agreement, 85% of tyrosine hydroxylase-positive cells remained in the nigral rAAV-MD-rGDNF group vs. only 49% in the lacZ group. A separate group of rats were given identical perinigral virus injections and were sacrificed at 3 and 10 weeks after surgery. Nigral GDNF protein expression remained relatively stable over the 10 weeks investigated. These data indicate that the use of rAAV, a noncytopathic viral vector, can promote delivery of functional levels of GDNF in a degenerative model of Parkinson's disease. PMID- 9391157 TI - Impaired motor coordination and persistent multiple climbing fiber innervation of cerebellar Purkinje cells in mice lacking Galphaq. AB - Mice lacking the alpha-subunit of the heterotrimeric guanine nucleotide binding protein Gq (Galphaq) are viable but suffer from ataxia with typical signs of motor discoordination. The anatomy of the cerebellum is not overtly disturbed, and excitatory synaptic transmission from parallel fibers to cerebellar Purkinje cells (PCs) and from climbing fibers (CFs) to PCs is functional. However, about 40% of adult Galphaq mutant PCs remain multiply innervated by CFs because of a defect in regression of supernumerary CFs in the third postnatal week. Evidence is provided suggesting that Galphaq is part of a signaling pathway that is involved in the elimination of multiple CF innervation during this period. PMID- 9391158 TI - Ca2+/calmodulin-binding peptides block phototransduction in Limulus ventral photoreceptors: evidence for direct inhibition of phospholipase C. AB - Phototransduction in Limulus photoreceptors involves a G protein-mediated activation of phospholipase C (PLC) and subsequent steps involving InsP3-mediated release of intracellular Ca2+. While exploring the role of calmodulin in this cascade, we found that intracellular injection of Ca2+/calmodulin-binding peptides (CCBPs) strongly inhibited the light response. By chemically exciting the cascade at various stages, we found the primary target of this effect was not in late stages of the cascade but rather at the level of G protein and PLC. That PLCdelta1 contains a calmodulin-like structure raised the possibility that PLC might be directly affected by CCBPs. To test this possibility, in vitro experiments were conducted on purified PLC. The activity of this enzyme was strongly inhibited by CCBPs and also inhibited by calmodulin itself. Our results suggest that the calmodulin-like region of PLC has an important role in regulating this enzyme. PMID- 9391159 TI - Adenylyl cyclase 6 is selectively regulated by protein kinase A phosphorylation in a region involved in Galphas stimulation. AB - Receptors activate adenylyl cyclases through the Galphas subunit. Previous studies from our laboratory have shown in certain cell types that express adenylyl cyclase 6 (AC6), heterologous desensitization included reduction of the capability of adenylyl cyclases to be stimulated by Galphas. Here we further analyze protein kinase A (PKA) effects on adenylyl cyclases. PKA treatment of recombinant AC6 in insect cell membranes results in a selective loss of stimulation by high (>10 nM) concentrations of Galphas. Similar treatment of AC1 or AC2 did not affect Galphas stimulation. Conversion of Ser-674 in AC6 to an Ala blocks PKA phosphorylation and PKA-mediated loss of Galphas stimulation. A peptide encoding the region 660-682 of AC6 blocks stimulation of AC6 and AC2 by high concentrations of Galphas. Substitution of Ser-674 to Asp in the peptide renders the peptide ineffective, indicating that the region 660-682 of AC6 is involved in regulation of signal transfer from Galphas. This region contains a conserved motif present in most adenylyl cyclases; however, the PKA phosphorylation site is unique to members of the AC6 family. These observations suggest a mechanism of how isoform selective regulatory diversity can be obtained within conserved regions involved in signal communication. PMID- 9391160 TI - Molecular determinants of tissue selectivity in estrogen receptor modulators. AB - Interaction of the estrogen receptor/ligand complex with a DNA estrogen response element is known to regulate gene transcription. In turn, specific conformations of the receptor-ligand complex have been postulated to influence unique subsets of estrogen-responsive genes resulting in differential modulation and, ultimately, tissue-selective outcomes. The estrogen receptor ligands raloxifene and tamoxifen have demonstrated such tissue-specific estrogen agonist/antagonist effects. Both agents antagonize the effects of estrogen on mammary tissue while mimicking the actions of estrogen on bone. However, tamoxifen induces significant stimulation of uterine tissue whereas raloxifene does not. We postulate that structural differences between raloxifene and tamoxifen may influence the conformations of their respective receptor/ligand complexes, thereby affecting which estrogen-responsive genes are modulated in various tissues. These structural differences are 4-fold: (A) the presence of phenolic hydroxyls, (B) different substituents on the basic amine, (C) incorporation of the stilbene moiety into a cyclic benzothiophene framework, and (D) the imposition of a carbonyl "hinge" between the basic amine-containing side chain and the olefin. A series of raloxifene analogs that separately exemplify each of these differences have been prepared and evaluated in a series of in vitro and in vivo assays. This strategy has resulted in the development of a pharmacophore model that attributes the differences in effects on the uterus between raloxifene and tamoxifen to a low-energy conformational preference imparting an orthogonal orientation of the basic side chain with respect to the stilbene plane. This three-dimensional array is dictated by a single carbon atom in the hinge region of raloxifene. These data indicate that differences in tissue selective actions among benzothiophene and triarylethylene estrogen receptor modulators can be ascribed to discrete ligand conformations. PMID- 9391161 TI - Pharmacological and immunohistochemical evidence for a functional nitric oxide synthase system in rat peritoneal eosinophils. AB - Eosinophil migration in vivo is markedly attenuated in rats treated chronically with the NO synthase (NOS) inhibitor Nomega-nitro-L-arginine methyl ester (L NAME). In this study, we investigated the existence of a NOS system in eosinophils. Our results demonstrated that rat peritoneal eosinophils strongly express both type II (30.2 +/- 11.6% of counted cells) and type III (24.7 +/- 7.4% of counted cells) NOS, as detected by immunohistochemistry using affinity purified mouse mAbs. Eosinophil migration in vitro was evaluated by using 48-well microchemotaxis chambers and the chemotactic agents used were N-formyl-methionyl leucyl-phenylalanine (fMLP, 5 x 10(-8) M) and leukotriene B4 (LTB4, 10(-8) M). L NAME (but not D-NAME) significantly inhibited the eosinophil migration induced by both fMLP (54% reduction for 1.0 mM; P < 0.05) and LTB4 (61% reduction for 1.0 mM; P < 0.05). In addition, the type II NOS inhibitor 2-amino-5,6-dihydro-6 methyl-4H-1,3-thiazine and the type I/II NOS inhibitor 1-(2 trifluoromethylphenyl) imidazole also markedly (P < 0. 05) attenuated fMLP- (52% and 38% reduction for 1.0 mM, respectively) and LTB4- (52% and 51% reduction for 1.0 mM, respectively) induced migration. The inhibition of eosinophil migration by L-NAME was mimicked by the soluble guanylate cyclase inhibitor 1H-[1,2,4] oxadiazolo [4,3,-a] quinoxalin-1-one (0.01 and 0.1 mM) and reversed by either sodium nitroprusside (0.1 mM) or dibutyryl cyclic GMP (1 mM). We conclude that eosinophils do express NO synthase(s) and that nitric oxide plays an essential role in eosinophil locomotion by acting through a cyclic GMP transduction mechanism. PMID- 9391162 TI - Unique allosteric regulation of 5-hydroxytryptamine receptor-mediated signal transduction by oleamide. AB - The effects of oleamide, an amidated lipid isolated from the cerebrospinal fluid of sleep-deprived cats, on serotonin receptor-mediated responses were investigated in cultured mammalian cells. In rat P11 cells, which endogenously express the 5-hydroxytryptamine2A (5HT2A) receptor, oleamide significantly potentiated 5HT-induced phosphoinositide hydrolysis. In HeLa cells expressing the 5HT7 receptor subtype, oleamide caused a concentration-dependent increase in cAMP accumulation but with lower efficacy than that observed by 5HT. This effect was not observed in untransfected HeLa cells. Clozapine did not prevent the increase in cAMP elicited by oleamide, and ketanserin caused an approximately 65% decrease. In the presence of 5HT, oleamide had the opposite effect on cAMP, causing insurmountable antagonism of the concentration-effect curve to 5HT, but had no effect on cAMP levels elicited by isoproterenol or forskolin. These results indicate that oleamide can modulate 5HT-mediated signal transduction at different subtypes of mammalian 5HT receptors. Additionally, our data indicate that oleamide acts at an apparent allosteric site on the 5HT7 receptor and elicits functional responses via activation of this site. This represents a unique mechanism of activation for 5HT G protein-coupled receptors and suggests that G protein-coupled neurotransmitter receptors may act like their iontropic counterparts (i.e., gamma-aminobutyric acid type A receptors) in that there may be several binding sites on the receptor that regulate functional activity with varying efficacies. PMID- 9391163 TI - Selection of peptides that functionally replace a zinc finger in the Sp1 transcription factor by using a yeast combinatorial library. AB - We have developed a strategy for the identification of peptides able to functionally replace a zinc finger domain in a transcription factor. This strategy could have important ramifications for basic research on gene regulation and for the development of therapeutic agents. In this study in yeast, we expressed chimeric proteins that included a random peptide combinatorial library in association with two zinc finger domains and a transactivating domain. The library was screened for chimeric proteins capable of activating transcription from a target sequence in the upstream regulatory regions of selectable or reporter genes. In a screen of approximately 1.5 x 10(7) transformants we identified 30 chimeric proteins that exhibited transcriptional activation, some of which were able to discriminate between wild-type and mutant DNA targets. Chimeric library proteins expressed as glutathione S-transferase fusions bound to double-stranded oligonucleotides containing the target sequence, suggesting that the chimeras bind directly to DNA. Surprisingly, none of the peptides identified resembled a zinc finger or other well-known transcription factor DNA binding domain. PMID- 9391164 TI - Sodium channel selectivity filter regulates antiarrhythmic drug binding. AB - Local anesthetic antiarrhythmic drugs block Na+ channels and have important clinical uses. However, the molecular mechanism by which these drugs block the channel has not been established. The family of drugs is characterized by having an ionizable amino group and a hydrophobic tail. We hypothesized that the charged amino group of the drug may interact with charged residues in the channel's selectivity filter. Mutation of the putative domain III selectivity filter residue of the adult rat skeletal muscle Na+ channel (micro1) K1237E increased resting lidocaine block, but no change was observed in block by neutral analogs of lidocaine. An intermediate effect on the lidocaine block resulted from K1237S and there was no effect from K1237R, implying an electrostatic effect of Lys. Mutation of the other selectivity residues, D400A (domain I), E755A (domain II), and A1529D (domain IV) allowed block by externally applied quaternary membrane impermeant derivatives of lidocaine (QX314 and QX222) and accelerated recovery from block by internal QX314. Neo-saxitoxin and tetrodotoxin, which occlude the channel pore, reduced the amount of QX314 bound in D400A and A1529D, respectively. Block by outside QX314 in E755A was inhibited by mutation of residues in transmembrane segment S6 of domain IV that are thought to be part of an internal binding site. The results demonstrate that the Na+ channel selectivity filter is involved in interactions with the hydrophilic part of the drugs, and it normally limits extracellular access to and escape from their binding site just within the selectivity filter. Participation of the selectivity ring in antiarrhythmic drug binding and access locates this structure adjacent to the S6 segment. PMID- 9391165 TI - Multi-responsiveness of single anterior pituitary cells to hypothalamic-releasing hormones: a cellular basis for paradoxical secretion. AB - The classic view for hypothalamic regulation of anterior pituitary (AP) hormone secretion holds that release of each AP hormone is controlled specifically by a corresponding hypothalamic-releasing hormone (HRH). In this scenario, binding of a given HRH (thyrotropin-, growth hormone-, corticotropin-, and luteinizing hormone-releasing hormones) to specific receptors in its target cell increases the concentration of cytosolic Ca2+ ([Ca2+]i), thereby selectively stimulating the release of the appropriate hormone. However, "paradoxical" responses of AP cells to the four well-established HRHs have been observed repeatedly with both in vivo and in vitro systems, raising the possibility of functional overlap between the different AP cell types. To explore this possibility, we evaluated the effects of HRHs on [Ca2+]i in single AP cells identified immunocytochemically by the hormone they stored. We found that each of the five major AP cell types contained discrete subpopulations that were able to respond to several HRHs. The relative abundance of these multi-responsive cells was 59% for lactotropes, 33% for thyrotropes, and in the range of 47-55% for gonadotropes, corticotropes, and somatotropes. Analysis of prolactin release from single living cells revealed that each of the four HRHs tested were able to induce hormone release from a discrete lactotrope subpopulation, the size of which corresponded closely to that in which [Ca2+]i changes were induced by the same secretagogues. When viewed as a whole, our diverse functional measurements of multi-responsiveness suggest that hypothalamic control of pituitary function is more complicated than previously envisioned. Moreover, they provide a cellular basis for the so-called "paradoxical" behavior of pituitary cells to hypothalamic hypophysiotropic agents. PMID- 9391166 TI - Resveratrol, a polyphenolic compound found in grapes and wine, is an agonist for the estrogen receptor. AB - The phytochemical resveratrol, which is found in grapes and wine, has been reported to have a variety of anti-inflammatory, anti-platelet, and anti carcinogenic effects. Based on its structural similarity to diethylstilbestrol, a synthetic estrogen, we examined whether resveratrol might be a phytoestrogen. At concentrations (approximately 3-10 microM) comparable to those required for its other biological effects, resveratrol inhibited the binding of labeled estradiol to the estrogen receptor and it activated transcription of estrogen-responsive reporter genes transfected into human breast cancer cells. This transcriptional activation was estrogen receptor-dependent, required an estrogen response element in the reporter gene, and was inhibited by specific estrogen antagonists. In some cell types (e.g., MCF-7 cells), resveratrol functioned as a superagonist (i.e., produced a greater maximal transcriptional response than estradiol) whereas in others it produced activation equal to or less than that of estradiol. Resveratrol also increased the expression of native estrogen-regulated genes, and it stimulated the proliferation of estrogen-dependent T47D breast cancer cells. We conclude that resveratrol is a phytoestrogen and that it exhibits variable degrees of estrogen receptor agonism in different test systems. The estrogenic actions of resveratrol broaden the spectrum of its biological actions and may be relevant to the reported cardiovascular benefits of drinking wine. PMID- 9391168 TI - Phloem sap proteins from Cucurbita maxima and Ricinus communis have the capacity to traffic cell to cell through plasmodesmata. AB - In angiosperms, the functional enucleate sieve tube system of the phloem appears to be maintained by the surrounding companion cells. In this study, we tested the hypothesis that polypeptides present within the phloem sap traffic cell to cell from the companion cells, where they are synthesized, into the sieve tube via plasmodesmata. Coinjection of fluorescently labeled dextrans along with size fractionated Cucurbita maxima phloem proteins, ranging in size from 10 to 200 kDa, as well as injection of individual fluorescently labeled phloem proteins, provided unambiguous evidence that these proteins have the capacity to interact with mesophyll plasmodesmata in cucurbit cotyledons to induce an increase in size exclusion limit and traffic cell to cell. Plasmodesmal size exclusion limit increased to greater than 20 kDa, but less than 40 kDa, irrespective of the size of the injected protein, indicating that partial protein unfolding may be a requirement for transport. A threshold concentration in the 20-100 nM range was required for cell-to-cell transport indicating that phloem proteins have a high affinity for the mesophyll plasmodesmal binding site(s). Parallel experiments with glutaredoxin and cystatin, phloem sap proteins from Ricinus communis, established that these proteins can also traffic through cucurbit mesophyll plasmodesmata. These results are discussed in terms of the requirements for regulated protein trafficking between companion cells and the sieve tube system. As the threshold value for plasmodesmal transport of phloem sap proteins falls within the same range as many plant hormones, the possibility is discussed that some of these proteins may act as long-distance signaling molecules. PMID- 9391167 TI - Structural activity of a cloned potassium channel (ROMK1) monitored with the atomic force microscope: the "molecular-sandwich" technique. AB - The atomic force microscope (AFM) was used to continuously follow height changes of individual protein molecules exposed to physiological stimuli. A AFM tip was coated with ROMK1 (a cloned renal epithelial potassium channel known to be highly pH sensitive) and lowered onto atomically flat mica surface until the protein was sandwiched between AFM tip and mica. Because the AFM tip was an integral part of a highly flexible cantilever, any structural alterations of the sandwiched molecule were transmitted to the cantilever. This resulted in a distortion of the cantilever that was monitored by means of a laser beam. With this system it was possible to resolve vertical height changes in the ROMK1 protein of >/=0.2 nm (approximately 5% of the molecule's height) with a time resolution of >/=1 msec. When bathed in electrolyte solution that contained the catalytic subunit of protein kinase A and 0.1 mM ATP (conditions that activate the native ion channel), we found stochastically occurring height fluctuations in the ROMK1 molecule. These changes in height were pH-dependent, being greatest at pH 7.6, and lowering the pH (either by titration or by the application of CO2) reduced their magnitude. The data show that overall changes in shape of proteins occur stochastically and increase in size and frequency when the proteins are active. This AFM "molecular-sandwich" technique, called MOST, measures structural activity of proteins in real time and could prove useful for studies on the relationship between structure and function of proteins at the molecular level. PMID- 9391169 TI - Evidence for the existence of a sulfonylurea-receptor-like protein in plants: modulation of stomatal movements and guard cell potassium channels by sulfonylureas and potassium channel openers. AB - Limitation of water loss and control of gas exchange is accomplished in plant leaves via stomatal guard cells. Stomata open in response to light when an increase in guard cell turgor is triggered by ions and water influx across the plasma membrane. Recent evidence demonstrating the existence of ATP-binding cassette proteins in plants led us to analyze the effect of compounds known for their ability to modulate ATP-sensitive potassium channels (K-ATP) in animal cells. By using epidermal strip bioassays and whole-cell patch-clamp experiments with Vicia faba guard cell protoplasts, we describe a pharmacological profile that is specific for the outward K+ channel and very similar to the one described for ATP-sensitive potassium channels in mammalian cells. Tolbutamide and glibenclamide induced stomatal opening in bioassays and in patch-clamp experiments, a specific inhibition of the outward K+ channel by these compounds was observed. Conversely, application of potassium channel openers such as cromakalim or RP49356 triggered stomatal closure. An apparent competition between sulfonylureas and potassium channel openers occurred in bioassays, and outward potassium currents, previously inhibited by glibenclamide, were partially recovered after application of cromakalim. By using an expressed sequence tag clone from an Arabidopsis thaliana homologue of the sulfonylurea receptor, a 7-kb transcript was detected by Northern blot analysis in guard cells and other tissues. Beside the molecular evidence recently obtained for the expression of ATP-binding cassette protein transcripts in plants, these results give pharmacological support to the presence of a sulfonylurea-receptor-like protein in the guard-cell plasma membrane tightly involved in the outward potassium channel regulation during stomatal movements. PMID- 9391170 TI - The roles of specific xanthophylls in photoprotection. AB - Xanthophyll pigments have critical structural and functional roles in the photosynthetic light-harvesting complexes of algae and vascular plants. Genetic dissection of xanthophyll metabolism in the green alga Chlamydomonas reinhardtii revealed functions for specific xanthophylls in the nonradiative dissipation of excess absorbed light energy, measured as nonphotochemical quenching of chlorophyll fluorescence. Mutants with a defect in either the alpha- or beta branch of carotenoid biosynthesis exhibited less nonphotochemical quenching but were still able to tolerate high light. In contrast, a double mutant that was defective in the synthesis of lutein, loroxanthin (alpha-carotene branch), zeaxanthin, and antheraxanthin (beta-carotene branch) had almost no nonphotochemical quenching and was extremely sensitive to high light. These results strongly suggest that in addition to the xanthophyll cycle pigments (zeaxanthin and antheraxanthin), alpha-carotene-derived xanthophylls such as lutein, which are structural components of the subunits of the light-harvesting complexes, contribute to the dissipation of excess absorbed light energy and the protection of plants from photo-oxidative damage. PMID- 9391171 TI - Chlorophyll precursors are signals of chloroplast origin involved in light induction of nuclear heat-shock genes. AB - Coordination between the activities of organelles and the nucleus requires the exchange of signals. Using Chlamydomonas, we provide evidence that plastid derived chlorophyll precursors may replace light in the induction of two nuclear heat-shock genes (HSP70A and HSP70B) and thus qualify as plastidic signal. Mutants defective in the synthesis of Mg-protoporphyrin IX were no longer inducible by light. Feeding of Mg-protoporphyrin IX or its dimethyl ester to wild type or mutant cells in the dark resulted in induction. The analysis of HSP70A promoter mutants that do or do not respond to light revealed that these chlorophyll precursors specifically activate the light signaling pathway. Activation of gene expression was not observed when protoporphyrin IX, protochlorophyllide, or chlorophyllide were added. A specific interaction of defined chlorophyll precursors with factor(s) that regulate nuclear gene expression is suggested. PMID- 9391172 TI - Structural model of cytochrome b559 in photosystem II based on a mutant with genetically fused subunits. AB - Photosystem II is a reaction center protein complex located in photosynthetic membranes of plants, algae, and cyanobacteria. Using light energy, photosystem II catalyzes the oxidation of water and the reduction of plastoquinone, resulting in the release of molecular oxygen. A key component of photosystem II is cytochrome b559, a membrane-embedded heme protein with an unknown function. The cytochrome is unusual in that a heme links two separate polypeptide subunits, alpha and beta, either as a heterodimer (alphabeta) or as two homodimers (alpha2 and beta2). To determine the structural organization of cytochrome b559 in the membrane, we used site-directed mutagenesis to fuse the coding regions of the two respective genes in the cyanobacterium Synechocystis sp. PCC 6803. In this construction, the C terminus of the alpha subunit (9 kDa) is attached to the N terminus of the beta subunit (5 kDa) to form a 14-kDa alphabeta fusion protein that is predicted to have two membrane-spanning alpha-helices with antiparallel orientations. Cells containing the alphabeta fusion protein grow photoautotrophically and assemble functional photosystem II complexes. Optical spectroscopy shows that the alphabeta fusion protein binds heme and is incorporated into photosystem II. These data support a structural model of cytochrome b559 in which one heme is coordinated to an alpha2 homodimer and a second heme is coordinated to a beta2 homodimer. In this model, each photosystem II complex contains two cytochrome b559 hemes, with the alpha2 heme located near the stromal side of the membrane and the beta2 heme located near the lumenal side. PMID- 9391174 TI - Adaptive evolution via a major gene effect: paedomorphosis in the Mexican axolotl. AB - Although adaptive evolution is thought to depend primarily on mutations of small effect, major gene effects may underlie many of the important differences observed among species in nature. The Mexican axolotl (Ambystoma mexicanum) has a derived mode of development that is characterized by metamorphic failure (paedomorphosis), an adaptation for an entirely aquatic life cycle. By using an interspecific crossing design and genetic linkage analysis, a major quantitative trait locus for expression of metamorphosis was identified in a local map of amplified fragment length polymorphisms. These data are consistent with a major gene hypothesis for the evolution of paedomorphosis in A. mexicanum. PMID- 9391173 TI - Plastid-localized acetyl-CoA carboxylase of bread wheat is encoded by a single gene on each of the three ancestral chromosome sets. AB - 5'-End fragments of two genes encoding plastid-localized acetyl-CoA carboxylase (ACCase; EC 6.4.1.2) of wheat (Triticum aestivum) were cloned and sequenced. The sequences of the two genes, Acc-1,1 and Acc-1,2, are 89% identical. Their exon sequences are 98% identical. The amino acid sequence of the biotin carboxylase domain encoded by Acc-1,1 and Acc-1,2 is 93% identical with the maize plastid ACCase but only 80-84% identical with the cytosolic ACCases from other plants and from wheat. Four overlapping fragments of cDNA covering the entire coding region were cloned by PCR and sequenced. The wheat plastid ACCase ORF contains 2,311 amino acids with a predicted molecular mass of 255 kDa. A putative transit peptide is present at the N terminus. Comparison of the genomic and cDNA sequences revealed introns at conserved sites found in the genes of other plant multifunctional ACCases, including two introns absent from the wheat cytosolic ACCase genes. Transcription start sites of the plastid ACCase genes were estimated from the longest cDNA clones obtained by 5'-RACE (rapid amplification of cDNA ends). The untranslated leader sequence encoded by the Acc-1 genes is at least 130-170 nucleotides long and is interrupted by an intron. Southern analysis indicates the presence of only one copy of the gene in each ancestral chromosome set. The gene maps near the telomere on the short arm of chromosomes 2A, 2B, and 2D. Identification of three different cDNAs, two corresponding to genes Acc-1,1 and Acc-1,2, indicates that all three genes are transcriptionally active. PMID- 9391175 TI - Is blindsight like normal, near-threshold vision? AB - Blindsight is the rare and paradoxical ability of some human subjects with occipital lobe brain damage to discriminate unseen stimuli in their clinically blind field defects when forced-choice procedures are used, implying that lesions of striate cortex produce a sharp dissociation between visual performance and visual awareness. Skeptics have argued that this is no different from the behavior of normal subjects at the lower limits of conscious vision, at which such dissociations could arise trivially by using different response criteria during clinical and forced-choice tests. We tested this claim explicitly by measuring the sensitivity of a hemianopic patient independently of his response criterion in yes-no and forced-choice detection tasks with the same stimulus and found that, unlike normal controls, his sensitivity was significantly higher during the forced-choice task. Thus, the dissociation by which blindsight is defined is not simply due to a difference in the patients' response bias between the two paradigms. This result implies that blindsight is unlike normal, near threshold vision and that information about the stimulus is processed in blindsighted patients in an unusual way. PMID- 9391176 TI - Spatial memory is related to hippocampal subcellular concentrations of calcium dependent protein kinase C isoforms in young and aged rats. AB - Relationships were examined between spatial learning and hippocampal concentrations of the alpha, beta2, and gamma isoforms of protein kinase C (PKC), an enzyme implicated in neuronal plasticity and memory formation. Concentrations of PKC were determined for individual 6-month-old (n = 13) and 24-month-old (n = 27) male Long-Evans rats trained in the water maze on a standard place-learning task and a transfer task designed for rapid acquisition. The results showed significant relationships between spatial learning and the amount of PKC among individual subjects, and those relationships differed according to age, isoform, and subcellular fraction. Among 6-month-old rats, those with the best spatial memory were those with the highest concentrations of PKCgamma in the particulate fraction and of PKCbeta2 in the soluble fraction. Aged rats had increased hippocampal PKCgamma concentrations in both subcellular fractions in comparison with young rats, and memory impairment was correlated with higher PKCgamma concentrations in the soluble fraction. No age difference or correlations with behavior were found for concentrations of PKCgamma in a comparison structure, the neostriatum, or for PKCalpha in the hippocampus. Relationships between spatial learning and hippocampal concentrations of calcium-dependent PKC are isoform specific. Moreover, age-related spatial memory impairment is associated with altered subcellular concentrations of PKCgamma and may be indicative of deficient signal transduction and neuronal plasticity in the hippocampal formation. PMID- 9391177 TI - A model of long-term memory storage in the cerebellar cortex: a possible role for plasticity at parallel fiber synapses onto stellate/basket interneurons. AB - By evoking changes in climbing fiber activity, movement errors are thought to modify synapses from parallel fibers onto Purkinje cells (pf*Pkj) so as to improve subsequent motor performance. Theoretical arguments suggest there is an intrinsic tradeoff, however, between motor adaptation and long-term storage. Assuming a baseline rate of motor errors is always present, then repeated performance of any learned movement will generate a series of climbing fiber mediated corrections. By reshuffling the synaptic weights responsible for any given movement, such corrections will degrade the memories for other learned movements stored in overlapping sets of synapses. The present paper shows that long-term storage can be accomplished by a second site of plasticity at synapses from parallel fibers onto stellate/basket interneurons (pf*St/Bk). Plasticity at pf*St/Bk synapses can be insulated from ongoing fluctuations in climbing fiber activity by assuming that changes in pf*St/Bk synapses occur only after changes in pf*Pkj synapses have built up to a threshold level. Although climbing fiber dependent plasticity at pf*Pkj synapses allows for the exploration of novel motor strategies in response to changing environmental conditions, plasticity at pf*St/Bk synapses transfers successful strategies to stable long-term storage. To quantify this hypothesis, both sites of plasticity are incorporated into a dynamical model of the cerebellar cortex and its interactions with the inferior olive. When used to simulate idealized motor conditioning trials, the model predicts that plasticity develops first at pf*Pkj synapses, but with additional training is transferred to pf*St/Bk synapses for long-term storage. PMID- 9391179 TI - Radionuclide therapy dose calculations: what accuracy can be achieved? PMID- 9391178 TI - Recovery of hearing and vocal behavior after hair-cell regeneration. AB - Postmitotic hair-cell regeneration in the inner ear of birds provides an opportunity to study the effect of renewed auditory input on auditory perception, vocal production, and vocal learning in a vertebrate. We used behavioral conditioning to test both perception and vocal production in a small Australian parrot, the budgerigar. Results show that both auditory perception and vocal production are disrupted when hair cells are damaged or lost but that these behaviors return to near normal over time. Precision in vocal production completely recovers well before recovery of full auditory function. These results may have particular relevance for understanding the relation between hearing loss and human speech production especially where there is consideration of an auditory prosthetic device. The present results show, at least for a bird, that even limited recovery of auditory input soon after deafening can support full recovery of vocal precision. PMID- 9391180 TI - Outpatient management of patients with large multinodular goitres treated with fractionated radioiodine. AB - The efficacy of fractionated out-patient radioiodine therapy in 38 patients with compressive symptoms due to long-standing large multinodular goitres was assessed. The diagnosis was established by clinical assessment in addition to technetium-99m pertechnetate thyroid scan or computed tomography scan of the thyroid and mediastinum. Oral iodine-131 therapy was administered as a 2.22 GBq (60 mCi) cumulative dose over 4 months (555 MBq per month). All patients were monitored with serum thyroid-stimulating hormone and free thyroxine (+/- free tri iodothyronine) assays before the treatment and after each dose fraction. Clinical and biochemical follow-up was performed on all patients and ranged from 6 to 45 months after therapy. The patients consisted of 35 female and three male patients with a median age of 59 years (range 37-87 years). Prior to treatment 20 patients were biochemically hyperthyroid and 18 were euthyroid. Overall, 71% of patients reported a subjective improvement in compressive symptoms and 29% reported no change. Clinically assessed reduction in goitre size occurred in 92% of patients while there was no change in 8%. At 3 months of follow-up, 31% of patients had become hypothyroid and at 18 months 66% were hypothyroid. Seven hyperthyroid patients (35%) became euthyroid and 13 hyperthyroid patients (65%) became hypothyroid. Three patients who became hypothyroid experienced neck soreness (transient in one patient, persistent in two patients). There were no differences in outcome between patients who were hyperthyroid and those who were euthyroid prior to treatment. Fractionated out-patient radioiodine therapy showed excellent short- and medium-term safety, was very well tolerated and offered a satisfactory alternative treatment to surgery. PMID- 9391181 TI - Thyroid volume measurement in thyrotoxic patients: comparison between ultrasonography and iodine-124 positron emission tomography. AB - The aim of this paper was to compare ultrasound (US) assessment of thyroid volume with that obtained by positron emission tomography (PET), in patients scheduled for adaptive radioiodine therapy, in which 50 Gy was prescribed to the functional PET volume. Firstly a pilot study was performed to ascertain the optimum method for US assessment of thyroid volume. Then 17 comparative measurements of thyroid volume by US and PET were made on 15 patients (two male and thirteen female, ages 28-73 years) with suspected Graves' disease. This comparison showed that in normal sized and enlarged thyroid glands (n=13), the ratio of functional PET to anatomical US volume was approximately 2:3. However, using the same ellipsoid model, PET and US assessment of anatomical volume agreed within the measurement errors. Owing to the presence of nodules and non-uniform distribution of radioiodine, the functional PET volume and anatomical US volume are often not equivalent. If high-resolution emission tomography (e.g. PET) is unavailable, the comparative data presented in this paper could be used to derive the functional volume from the US volume for calculating functional thyroid dose in hyperthyroid patients undergoing radioiodine therapy. PMID- 9391182 TI - Extraction and retention of technetium-99m Q12, technetium-99m sestamibi, and thallium-201 in isolated rat heart during coronary acidemia. AB - Technetium-99m Q12 and 99mTc-sestamibi are cationic lipophilic myocardial perfusion imaging tracers. Because myocardium in areas of ischemia becomes acidotic, experiments were designed to differentiate the effects of myocardial perfusate pH on radiotracer extraction and retention independent of substrate availability. We hypothesized that 99mTc-Q12 and 99mTc-sestamibi single-pass uptake and retention would be unaffected by a modest reduction in coronary perfusate pH. Isolated rat hearts were perfused at constant flow with Krebs Henseleit buffer enriched with bovine red blood cells (20%). The indicator dilution method was used to measure the maximum extraction (Emax) and net extraction (Enet) of thallium-201 and 99mTc-Q12 (n = 8) or 201Tl and 99mTc sestamibi (n = 7) during baseline perfusion (pH = 7.4), during acidemic (pH = 6.7) perfusion, and during a restitution period with normal perfusate (pH = 7.4). 201Tl Emax (0.71+/-0.03) was greater than either 99mTc-Q12 or 99mTc-sestamibi Emax (0.27+/-0.02 and 0.26+/-0.01 respectively, P<0.0001). Acidemia significantly reduced 201Tl Emax (0.65+/-0.03, P<0.02) but not 99mTc-Q12 or 99mTc-sestamibi Emax (0.25+/-0.02 and 0.24+/-0.02 respectively). During control perfusion Enet of 201Tl was greater than that of 99mTc-Q12 at 3 and 5 min and greater than that of 99mTc-sestamibi at 3 min. 99mTc-Q12 Enet was less than 99mTc-sestamibi Enet at 3, 5, and 10 min. Acidemia decreased 201Tl and 99mTc-sestamibi Enet at 3, 5, and 10 min but had no effect on 99mTc-Q12 Enet. It is concluded that Emax of 99mTc-Q12 is less than that of 201Tl but is not different from that of 99mTc-sestamibi. Enet of 99mTc-Q12 is less than that of 99mTc-sestamibi. PMID- 9391183 TI - Comparison of flow capacities of arterial and venous grafts for coronary artery bypass grafting: evaluation with exercise thallium-201 single-photon emission tomography. AB - Stress thallium-201 tomography was performed to compare the flow capacities of arterial and saphenous vein grafts in patients with coronary artery bypass grafting (CABG). One hundred and seven consecutive patients (95 male and 12 female; mean age 58+/-9.1 years) underwent exercise-redistribution 201Tl myocardial single-photon emission tomography 4-5 weeks after CABG. When a reversible perfusion defect was present in the area covered by a patent bypass graft, the flow capacity of the graft was defined as insufficient. Of all 285 grafts, 211 were considered as complete bypass. Reversible perfusion defects were present in 29 (27%) of 108 myocardial areas supplied by patent arterial grafts but in only 5 (5%) of 103 myocardial areas supplied by patent saphenous vein grafts (P<0.0001). In the LAD area reversible defects were observed in 22 of 82 areas covered by arterial grafts, in contrast to only 1 of 29 areas covered by venous grafts (P<0.01); in the RCA area reversible defects were observed in 7 of 17 and 4 of 41 areas respectively (P<0.01). There was no difference between the native coronary artery stenosis bypassed by patent arterial and venous grafts (88%+/-12% vs 86%+/-14% respectively, P=0.27). In conclusion, flow capacities during peak myocardial demand were more frequently insufficient in arterial bypass grafts than in saphenous vein grafts. PMID- 9391184 TI - Preoperative parathyroid gland localization with technetium-99m sestamibi in secondary hyperparathyroidism. AB - Technetium-99m sestamibi scintigraphy has become a valuable tool in locating parathyroid glands in patients with primary hyperparathyroidism. The aim of this study was to evaluate its usefulness in secondary hyperparathyroidism. Twenty patients were injected intravenously with 740 MBq of 99mTc-sestamibi and images were obtained at 15 min and 2 h post injection. All patients underwent parathyroid ultrasonography (US) as well as bilateral surgical neck exploration and 64 parathyroid glands were removed. US revealed at least one enlarged gland in 15/20 patients (75%), while 99mTc-sestamibi scintigraphy showed focal areas of increased uptake in at least one gland in 17/20 patients (85%). When imaging results for all glands were evaluated according to surgical results, sensitivity was 54% for parathyroid scintigraphy and 41% for US, and specificity was 89% for both imaging techniques. There was a discrepancy between the two imaging modalities in 28 glands (35%). The mean surgical weight of US-positive glands (1492+/-1436 mg) was significantly higher than that of US-negative glands (775+/ 703 mg) (P<0.05). However, there were no significant differences in weight between sestamibi-positive and sestamibi-negative glands. When only sestamibi positive glands were considered, a positive correlation between uptake and weight was found (r=0.4, P<0.05). In conclusion, parathyroid US and 99mTc-sestamibi scintigraphy are complementary imaging techniques in the preoperative localization of abnormal parathyroid glands in patients with secondary hyperparathyroidism. The limited sensitivity of the techniques means that patients will still require bilateral neck exploration; therefore routine preoperative parathyroid scanning in renal patients is not justified. PMID- 9391185 TI - Revisiting the prognostic role of gallium scintigraphy in low-grade non-Hodgkin's lymphoma. AB - The purpose of this study was threefold: to evaluate the role of gallium-67 scintigraphy in the staging of low-grade non-Hodgkin's lymphomas (LGNHL), to assess the relationship between the expression of CD71 on the surface of the neoplastic cells and the 67Ga uptake by the tumour, and to establish the contribution of 67Ga scan in defining the prognosis of LGNHL. Forty-eight patients with untreated LGNHL diagnosed in a single institution over a decade were reviewed. The end point of the study was survival of the patients according to the scintigraphic 67Ga score at diagnosis. In addition to 67Ga scan, other prognostic variables were studied, relating to the neoplastic burden, the biology of the tumour and the host. Univariate and multivariate analyses were used. 67Ga scan identified only 116/286 (41%) nodes involved by lymphoma that were detected by clinical examination or computed tomography scan. A scintigraphic scoring system with an arbitrary cut-off value of 3 (high scan score) was able to predict patients with a dismal prognosis: with a mean follow-up of 47 months (range: 1 146 months) the median survival time was 28 months in patients with a high scan score and 74 months in patients with a low scan score (P=0.002). CD71 values were 27. 4%+/-14.9% (mean +/-SD) in the former and 8.9%+/-7.2% in the latter (P=0.0001). Only performance status and extranodal sites were significant variables for prognosis in multivariate analysis. It is concluded that 67Ga scan is inaccurate in staging but might be very important in defining the prognosis in LGNHL, in association with other prognostic variables. PMID- 9391186 TI - Semi-quantitative assessment of cerebral blood flow with 99mTc-HMPAO SPET in type I diabetic patients with no clinical history of cerebrovascular disease. AB - In 65 type I diabetic patients we prospectively evaluated brain perfusion by means of single-photon emission tomography after the injection of 740- 1110 MBq of technetium-99m hexamethylpropylene amine oxime. Thirty-five of the patients presented complications secondary to their diabetes. None showed CNS symptoms. A semiquantitative analysis was performed drawing 50 symmetrical regions of interest (ROIs) per patient. The relative contribution of each ROI to the total blood flow in each slice was compared with the relative contribution of the same ROI in a control group of ten healthy volunteers. Relative values of any ROI in the study group higher or lower than the mean +/-2 SD in respect of the same ROI in the control group were considered abnormal. The results revealed hypoperfusion in 207 ROIs in the 65 patients with diabetes mellitus: of these ROIs, 113 were frontal, 10 frontotemporal, 20 temporal, 18 parietal, 11 occipital and 35 cerebellar. A total of 137 ROIs showed hyperperfusion: 17 frontal, 3 frontotemporal, 19 temporal, 18 parietal, 19 parieto-occipital, 29 occipital and 32 cerebellar. Out of 65 type I diabetic patients, 61 showed at least one hypoperfused ROI (P = 0.0064 vs. controls) and 25 showed more than three hypoperfused ROIs. None of the control subjects showed more than three hypoperfused regions (P<0.001). The results obtained demonstrate the existence of subclinical abnormalities of brain blood perfusion in patients with type I diabetes mellitus and no history of cerebrovascular disease, thereby allowing the initiation of intensive preventive measures. PMID- 9391187 TI - Calculation of residence times and radiation doses using the standard PC software Excel. AB - We developed a program which aims to facilitate the calculation of radiation doses to single organs and the whole body. IMEDOSE uses Excel to include calculations, graphical displays, and interactions with the user in a single general-purpose PC software tool. To start the procedure the input data are copied into a spreadsheet. They must represent percentage uptake values of several organs derived from measurements in animals or humans. To extrapolate these data up to seven half-lives of the radionuclide, fitting to one or two exponentional functions is included and can be checked by the user. By means of the approximate time-activity information the cumulated activity or residence times are calculated. Finally these data are combined with the absorbed fraction doses (S-values) given by MIRD pamphlet No. 11 to yield radiation doses, the effective dose equivalent and the effective dose. These results are presented in a final table. Interactions are realized with push-buttons and drop-down menus. Calculations use the Visual Basic tool of Excel. In order to test our program, biodistribution data of fluorine-18 fluorodeoxyglucose were taken from the literature (Meija et al., J Nucl Med 1991; 32:699-706). For a 70-kg adult the resulting radiation doses of all target organs listed in MIRD 11 were different from the ICRP 53 values by 1%+/-18% on the average. When the residence times were introduced into MIRDOSE3 (Stabin, J Nucl Med 1996; 37:538-546) the mean difference between our results and those of MIRDOSE3 was -3%+/-6%. Both outcomes indicate the validity of the present approach. PMID- 9391188 TI - The role of technetium-99m sestamibi single photon emission tomography in the follow-up of malignant melanoma and the detection of lymph node metastases. AB - In the follow-up of patients with malignant melanoma treated by surgical resection of the cutaneous tumour, it is important to achieve early detection of possible lymph node metastasis. In many cases, clinical examination alone will not be sufficient. In our study, single-photon emission tomography (SPET) with technetium-99m sestamibi (MIBI) was used in the assessment of 30 patients with previously resected malignant melanoma when the clinical examination raised the suspicion of lymph node metastasis. Using MIBI, 16 out of 17 lymph node metastases were detected and confirmed by histology. No false-positive results were obtained during this prospective study. It is concluded that MIBI scintigraphy may be useful in the early detection of lymph node metastases of malignant melanomas. If our preliminary results are confirmed, early detection of lymph node metastasis of previously resected malignant melanoma by 99mTc-MIBI scintigraphy may have a significant impact on the management of these patients. PMID- 9391189 TI - Highlights of the annual meeting of the European Association of Nuclear Medicine: Glasgow 1997. The flowering of science and art of nuclear medicine in Europe. PMID- 9391190 TI - Radiation dose rates from patients receiving iodine-131 therapy for carcinoma of the thyroid PMID- 9391191 TI - Rare infections in pediatric surgery PMID- 9391193 TI - Pyogenic liver abscess complicating a ventriculoperitoneal shunt. AB - Pyogenic liver abscess is a rare complication of ventriculoperitoneal (VP) shunting. We report a 4-month-old female with this complication who was successfully treat ed by computed tomography-guided percutaneous transhepatic catheter drainage, shunt externalization, and parenteral antibiotics. Liver abscess is a possible intra-abdominal complication of VP shunting, and imaging studies are good adjuncts in making the clinical diagnosis. PMID- 9391192 TI - Surgical aspects of an outbreak of Yersinia enterocolitis. AB - Eleven patients with Yersinia enterocolitica infections were identified in the Upper Valley of New Hampshire and Vermont during October and November of 1995. Three children presented with an appendicitis-like picture. Two underwent appendectomy, one of whom was the outbreak's index case. Both appendectomy patients presented with lower abdominal pain, fever, vomiting, and a right lower quadrant mass associated with leukocytosis. Both had terminal ileitis, and in both, cultures of peritoneal fluid and a mesenteric lymph node grew Y. enterocolitica. Even during an outbreak there is no consistently reliable nonoperative way to separate a sporadic case of appendicitis from one whose appendicitis-like symptoms are due to Yersinia. In addition, a small percentage of Yersinia patients will present with true appendicitis as a complication of their disease. PMID- 9391194 TI - Hepatic duct stone associated with chlamydia sepsis: a rare condition in childhood. AB - Stone formation in the biliary system is a rare condition in infants. A few cases of bile stones in the biliary tree have been reported with underlying predisposing factors, such as sepsis and antibiotic usage. This article describes a surgically treated 16-week-old infant with recurrent cholangitis who had a bile stone in the hepatic duct after chlamydia sepsis. PMID- 9391195 TI - Lung resection in pediatric patients. AB - An evaluation of all pediatric patients with primary or secondary pulmonary disease operated upon from January 1993 to July 1996 by the same senior surgeon was carried out. The inclusion criterion was a lung resection in patients aged less than 14 years. Children were divided into two categories according to the neoplastic or non-neoplastic nature of their disease. In the first group a lobectomy was performed for primary lesions and wedge resection for secondary ones. In the second group lobar emphysema and cystic dysplasia were the major indications for lobectomy, while diagnostic wedge resections were performed for interstitial/infiltrative lesions. Several groups of techniques were identified according to the type of approach and the suture method. Video-assisted thoracoscopic surgery and a muscle-sparing approach were compared to classic posterolateral thoracotomy. The mechanical stapler-suturing method was compared to the manual suturing. Our results demonstrate the importance of mechanical suturing, particularly in decreasing anesthesia time and reducing the risk of dehiscence. The minimally invasive approach associated with mini-thoracotomy was particularly useful for patients with reduced oxygen saturation due to ventilatory and gas-exchange problems. The roles of staplers in lung parenchymal resection and minimally invasive procedures for improving the postoperative thoracic compliance of pediatric patients are stressed. PMID- 9391196 TI - The significance of biliary sludge in children with sickle cell disease. AB - The prevalence of cholelithiasis was studied prospectively by abdominal ultrasound (US) examination in 305 children with sickle cell disease aged 1-18 years (mean 10.45). Gallstones were present in 60 children (19.7%); an additional 50 had biliary sludge only (16.4%). On follow-up of 35 of the 50 children with sludge, 23 (65.7%) had developed gallstones and 5 had already had a cholecystectomy. Five continued to have sludge on follow-up while 7 were reported to have no sludge. Children with US evidence of sludge should be followed up regularly by US, and those who develop gallstones should undergo elective cholecystectomy. For those with biliary sludge only, we recommend elective cholecystectomy if there are hepatobiliary symptoms. PMID- 9391197 TI - Is a normally functioning gastrointestinal tract necessary for normal growth in late gestation? AB - It is known that neonates with congenital abnormalities of the intestine tend to be growth-retarded. We wished to explore the hypothesis that normal fetal gut function is needed for normal growth in late gestation. If this is true, then different populations of babies with different congenital gut abnormalities would be expected to have similar impairments of growth and be small at birth. This growth retardation would be more marked in term than in preterm babies and would be independent of other congenital anomalies. To test these hypotheses, we examined 43 babies born with gastroschisis (GS) in Auckland, New Zealand; 69 babies born with GS in Birmingham, England; and 60 babies born with intestinal atresia (IA) in Auckland. For Auckland babies with GS, the mean weight standard deviation score (WSDS) (i.e., birth weight relative to the mean birth weight for gestation) for term babies was lower than that for preterm babies (-0.932+/-0.180 vs -0.064+/-0.237, P=0.014). This was also true for Birmingham babies with GS ( 0.991+/-0.193 vs -0.36 +/-0.153, P=0.028). For babies with IA, the mean WSDS for term babies was lower than that for preterm babies (-0.627+/-0.266 vs 0. 057+/ 0.211, P=0.034). There was no significant difference between the mean WSDS of babies with and without major congenital abnormalities (-0.402+/-0.201 vs -0.271, P=0.70). Our results demonstrate that term babies born with GS are significantly growth-retarded compared with premature babies born with GS. Term babies born with a proximal IA are also growth-retarded. This strongly suggests that in late gestation, the normal growth is dependent on a normally functioning gastrointestinal tract that allows exposure of the proximal intestinal mucosa to ingested amniotic fluid. PMID- 9391198 TI - Laparoscopic versus conventional appendectomy in children. AB - Over a period of 6 months, 48 children who underwent conventional appendectomy (CA) were compared to 34 children who had laparoscopic appendectomy (LA) for acute and recurrent subacute appendicitis. Their ages ranged from 4 to 12 years (mean 8). LA took significantly longer, 76 min, versus 50 min for CA. Less than 2 days' hospitalization was required in 95% of LA cases and 45% of CA cases. Time to return to normal activity averaged 7 days for LA and 12 days for CA (P < 0.001). Wound complications were fewer and the cosmetic appearance was most satisfactory in LA patients. LA is a safe operation that has the advantage of being exploratory, with shorter hospitalization time, early ambulation, and superior cosmetic results. PMID- 9391199 TI - Long-term functional, manometric, and endosonographic evaluation of patients operated upon with the Duhamel technique. AB - Long-term functional results, anal endosonography (AES), and anal canal manometry were recorded in 48 patients aged 10 to 24 years (median 18) operated upon with the Duhamel technique for Hirschsprung's disease; 60.4% had perfect fecal control, 31.3% occasional staining and/or gas incontinence, and 8.3% constant fecal soiling, and 10.4% complained of constipation. Compared to normals, the patients had significantly reduced anal canal resting and squeeze pressures. AES visualized scar tissue in both the internal and external anal sphincter. PMID- 9391200 TI - Temporal stability of acetylcholinesterase staining in colonic and rectal neural tissue. AB - Confirmation of the clinical diagnosis of Hirschsprung's disease on standard rectal suction biopsy requires demonstration of aganglionosis in 60 adequate serial sections of submucosa. Positive staining for acetylcholinesterase (AChE), demonstrating an increase in nerve fibres within the lamina propria, muscularis mucosae, and subjacent submucosa, is a useful adjunctive test. In this study, sections of distal colonic muscularis propria and rectal mucosa were stained for AChE over a period of days following storage at 4 degrees C and at room temperature (15-20 degrees C). Positive staining of neural tissue was demonstrated in specimens stored at 4 degrees C for up to 14 days, at which time the experiment was discontinued due to tissue autolysis. Positive staining of the myenteric plexus in colonic specimens stored at room temperature also continued until tissue dissolution became marked at 5 days. This study has demonstrated stability of AChE staining of intestinal neural tissue in specimens stored at 4 degrees C for 14 days, which suggests that reliable staining for AChE should still be achievable if rectal biopsies are taken in clinics/hospitals without access to staining facilities, provided that tissues are transferred (fresh and moist, at 4 degrees C) to a reference laboratory for staining within several days of the biopsy procedure. PMID- 9391201 TI - Treatment of vesicoureteric reflux in a sheep model using subureteric injection of cultured fetal-bladder tissue. AB - To investigate the role of injecting cultured fetal-bladder tissue into the region of the vesicoureteric orifice (VUO) to correct surgically produced vesicoureteric reflux (VUR), 12 Coopworth ewe lambs were studied. Four weeks after incising the intravesical segment of ureter, VUR was demonstrated by micturating cystourethrography. Bladder tissue was obtained from a fetal Coopworth lamb at 10 weeks' gestation, cultured in RPMI 1640, and injected into the region of the VUO of 1 ureter of each lamb using an open surgical approach. The lambs were killed between 1 and 6 months after the injection. Smooth-muscle cells from the cultured fetal bladder tissue were identified by the monoclonal antibodies HHF-35 for muscle alpha-actin and D33 for muscle desmin, and by electron microscopy. One lamb died of a gastrointestinal infection at 8 weeks of age. Of the remaining 11 animals, the injection of fetal-bladder tissue corrected the reflux in 7, while it was reduced in degree in 3 and persisted unchanged in 1. The reflux on the contralateral control side was also corrected in 6 ureters and improved in 2. Using histochemical techniques, grafted fetal-bladder tissue could not be differentiated from host tissue in the region of the VUO. Histopathological studies failed to show any injected tissue in distant organs. This study has shown that surgically-induced VUR in lambs was corrected or improved by the injection of cultured fetal-bladder tissue into the submucosa adjacent to the VUO. PMID- 9391202 TI - Undescended testes: incidence in 1,002 consecutive male infants and outcome at 1 year of age. AB - In a study of 1,002 consecutive Malaysian male newborns, 48 (4.8%) were found to have undescended testes (UDT). The rate and laterality of the UDT were associated with lower birth weight (P < 0.001) and prematurity (P < 0.001). Boys with UDT were also more likely to have other congenital abnormalities of the external genitalia, the commonest being hydrocele. No correlation between UDT and maternal age, birth order, social class, or mode of delivery was demonstrated in this study. Although 26/34 (76.5%) of UDT achieved full spontaneous descent by 1 year of age, 1.1% of all infants whose testes remained undescended required regular long-term follow-up with surgical referral and correction at an appropriate time. A premature infant with UDT is more likely to achieve full testicular descent at 1 year of age than a term infant. PMID- 9391203 TI - Results of Wilms' tumour management in two tertiary-care hospitals in Asia. AB - In the period 1985-1995, 87 children underwent surgery for Wilms' tumour; 16 were lost to follow-up. Of the remaining children, 27 presented with stage I disease, 11 with stage II, 12 with stage III, 14 with stage IV, and 6 with stage V. One child was not staged. The histology was favourable Wilms' tumour in 44, anaplastic in 12, unclassified in 8, clear-cell sarcoma in 4, and rhabdoid tumour in 3. Although a total nephrectomy was generally performed, partial renal surgery was performed for 6 bilateral and 4 unilateral tumours, the latter including 2 fused kidneys. Preoperative chemotherapy was employed with benefit in massive tumours, tumour in fused kidneys, bilateral tumours, and preoperatively diagnosed inferior vena caval tumour thrombi. Postoperative chemotherapy, employed in all cases, consisted of actinomycin D and vincristine with the addition of adriamycin in anaplastic and advanced-stage tumours. Ten children underwent second-line chemotherapy for disease unresponsive to the above management, but only 1 of these is currently free of disease. Postoperative tumour-bed radiotherapy, used in selected cases, prevented local recurrence in stage I and II disease. However, 20% of stage I and II patients not receiving radiotherapy developed tumour-bed recurrence. Twenty-three children have died and 5 with advanced disease and incomplete follow-up are presumed to be dead. Nine children are currently on treatment; 34 have successfully completed treatment, the disease-free survival in stages I-V being 81%, 75%, 42%, 14%, and 50%, respectively. Overall disease-free survival was 69% for Wilms' tumour of favourable histology and 50% for anaplastic tumours. The 3 patients with rhabdoid tumours and 3 of 4 with clear-cell sarcomas have died. Wilms' tumour management in the developing world is compromised by cases lost to follow-up and late presentation with massive tumours and advanced stage. Preoperative chemotherapy is advantageous in a number of cases, and postoperative radiotherapy should be deployed more frequently. PMID- 9391204 TI - Leiomyoma of the esophagus and bronchus in a child. AB - Leiomyomas of the esophagus and/or the bronchus have rarely been reported in children. To our knowledge, the simultaneous presence of this tumor in both the esophagus and a bronchus in a child has not been previously reported. A 7-year old boy presented with respiratory and esophageal symptoms and was found to have a leiomyoma of the esophagus and right main bronchus. The esophageal leiomyoma was treated with limited myotomy, but bronchoscopic resection was possible for the bronchial lesion. The postoperative result was excellent, with normal swallowing and no residual respiratory problems at 1-year follow-up. PMID- 9391205 TI - Abdominal wall plasty for a premature infant with congenital diaphragmatic hernia. AB - This paper reports a premature infant with a congenital diaphragmatic hernia (CDH) who underwent an abdominal wall plasty to enlarge the abdominal cavity, one of twin infants born at 32 weeks weighing 1,255 g. After stabilization, the herniated viscera were reduced from the pleural cavity and the abdominal wall muscle and skin layers were replaced by a Gore-tex patch without closure of the diaphragmatic defect. Respiratory and circulatory conditions were stable during the perioperative period. Postoperatively, a roentogenogram showed expansion of the lung. However, his condition deteriorated 24 h after surgery, triggered by intratracheal suction, and he died on the 4th day of life despite the use of high frequency oscillation, catecholamines, and vasodilators. Postmortem examination showed severely hypoplastic lungs. Abdominal wall plasty may be a less invasive initial procedure, however, further studies, such as comparison with the standard method or conservative management, are needed using a large clinical group or animal models to justify the usefulness of this procedure. PMID- 9391206 TI - Pancreatic inflammatory pseudotumour: an uncommon childhood lesion mimicking a malignant tumour. AB - We describe a rare case of pancreatic inflammatory pseudotumour that clinically presented as a malignant tumour in an 11-year-old boy. We also review all six previously described cases, including three that occurred in children. Complete surgical removal is regarded as the best choice of therapy, and follow-up in our case showed no evidence of disease after 3 years. It is important to be aware of this lesion, since it can be confused with a malignant tumour both radiologically and clinically. PMID- 9391207 TI - A stercoraceous ulcer of the colon in neglected Hirschsprung's disease. AB - A large, nonspecific, chronic ulcer was found in the sigmoid colon of a 13-year old child with neglected, undiagnosed Hirschsprung's disease (HD). There is no known association between HD and colonic ulcers, suggesting that the ulcer was a true stercoraceous ulcer of the colon and not an intrinsic defect of the aganglionotic bowel. PMID- 9391208 TI - Two unusual cases of megacolons. AB - A 13-year-old boy with features of intestinal obstruction was found at laparotomy to have a sigmoid volvulus. A 7-year-old girl with a similar presentation had a tight stricture at the rectosigmoid junction causing obstruction. In both cases the proximal colon was grossly loaded with faeces. In the first child, a colostomy after resecting the sigmoid colon was considered the safer option, whereas in the second, an innovative method to decompress the proximal loaded colon using a sterilised colostomy bag was employed, enabling a primary anastomosis to be performed. PMID- 9391209 TI - Surgical management of complete ureteric duplication abnormalities. AB - In 2 decades (1974-1993), the senior author (S.A.) managed 148 patients with various abnormalities associated with complete ureteric duplication. Included were 72 patients with primary vesicoureteric reflux, 50 with ureteroceles, and 26 with upper-pole ectopic ureters. The majority of the patients were female, and the common clinical presentations included urinary tract infection (UTI), UTI with septicemia, and urinary incontinence. Ten cases were diagnosed after recognition of a renal abnormality on prenatal ultrasonography, an avenue that has provided new challenges, new opportunities, and new dilemmas. This review article is based on the authors' experience together with an analysis of current emphasis on early diagnosis, minimal surgery, and maximum preservation of renal function. PMID- 9391210 TI - Polyorchidism. AB - Polyorchidism is defined as the presence of more than two testes. We report the case of a 3-year-old boy and review the embryology and surgical management of the condition. PMID- 9391211 TI - Supernumerary ovary associated with Wilms' tumor. AB - Supernumerary ovaries are gynecological rarities. To the best of our knowledge, only 18 histologically confirmed cases have been documented so far, all reported in adult women. We present the first such case in a 5-year-old child, who also had a Wilms' tumor of the left kidney. PMID- 9391212 TI - Bilateral testicular tumors in an infant. AB - A 7-month-old infant showed bilateral enlarged, nontender scrotal masses. The level of alpha-fetoprotein was greater than 10,000 ng/ml preoperatively; a high left inguinal orchiectomy was performed for a suspected yolk-sac tumor. The right testis was diagnosed as a mature teratoma because it was not possible to establish a line of cleavage between the tumor and the normal tissue, and a high right inguinal orchiectomy was performed. Only one case of bilateral testicular teratomas has been reported in the literature to date. We report a rare second case of bilateral testicular tumors, one a yolk-sac tumor and the other a teratoma. PMID- 9391213 TI - Presacral teratoma presenting with congenital urinary ascites. AB - Congenital teratomas occur most frequently in the sacrococcygeal region. Most grown into a large perineo-sacral swelling that is conspicuous externally. Infrequently, the neoplasm is contained almost entirely within the pelvis in the presacral space. Congenital urinary ascites is observed in patients with obstructive uropathy; posterior urethral valves in a newborn is one of the most prominent causes of urinary ascites. We report a case of presacral teratoma leading to rupture of the urinary bladder due to outflow obstruction and causing urinary ascites. The ascites was drained, the bladder was repaired, and the teratoma was successfully excised. A review of the literature did not reveal any similar case. PMID- 9391214 TI - Telomere length, chromatin structure and chromosome fusigenic potential. AB - Telomeres are specialized structures at chromosome ends that are thought to function as buffers against chromosome fusion. Several studies suggest that telomere shortening may render chromosomes fusigenic. We used a novel quantitative fluorescence in situ hybridization procedure to estimate telomere length in individual mammalian chromosomes, and G-banding and chromosome painting techniques to determine chromosome fusigenic potential. All analysed Chinese hamster and mouse cell lines exhibited shorter telomeres at short chromosome arms than at long chromosome arms. However, no clear link between short telomeres and chromosome fusigenic potential was observed, i.e. frequencies of telomeric associations were higher in cell lines exhibiting longer telomeres. We speculate that chromosome fusigenic potential in mammalian cell lines may be determined not only by telomere length but also by the status of telomere chromatin structure. This is supported by the observed presence of chromatin filaments linking telomeres in Chinese hamster chromosomes and of multibranched chromosomes oriented end-to-end in the murine severe combined immunodeficient (SCID) cell line. Multibranched chromosomes are the hallmark of the human ICF (Immune deficiency, Centromeric instability, Facial abnormalities) syndrome, characterized by alterations in heterochromatin structure. PMID- 9391215 TI - The role of sister chromatid cohesiveness and structure in meiotic behaviour. AB - Sister chromatid cores, kinetochores and the connecting strand between sister kinetochores were differentially silver stained to analyse the behaviour of these structures during meiosis in normal and two spontaneous desynaptic individuals of Chorthippus jucundus (Orthoptera). In these desynaptic individuals most of the chromosomes appear as univalents and orient equationally in the first meiotic division. Despite this abnormal segregation pattern, the changes in chromosome structure follow the same timing as in normal individuals and seem to be strictly phase dependent. Chromosomes in the first prometaphase have associated sister kinetochores and sister chromatid cores that lie in the chromosome midline; we propose that this promotes the initial monopolar orientation of chromosomes. However, the requirements of tension for stable attachment to the spindle force the autosomal univalents to acquire amphitelic orientation. Sister kinetochores behave in a chromosome orientation-dependent manner and, in the first metaphase, they appear to be interconnected by a strand that can be detected by silver impregnation, as seen in the second metaphase of wild-type individuals. The disappearance of the sister kinetochore-connecting strand, needed for equational chromatid segregation, however, can only take place in the second meiotic division. This connecting strand is ultimately responsible for the inability of chromosomes to segregate sister chromatids in the first anaphase. PMID- 9391216 TI - Immunocytochemical visualization of the centromeres during male and female meiosis in Lilium longiflorum. AB - Immunofluorescence staining with an antiserum raised against a presumptive meiotic histone, which has been shown to appear prior to male meiosis in liliaceous plants, preferentially stained the centromere (kinetochore) region of meiotic chromosomes in microsporocytes and megasporocytes. Using this antiserum, we were able clearly to visualize the centromeres at all important meiotic stages in microsporocytes, namely, the association and fusion of centromeres of homologous chromosomes at zygotene-pachytene in prophase I, the disjunction of the homologous centromeres at diplotene, the doubling of each centromere at metaphase I and nonseparation of the sister centromeres at anaphase I, by confocal laser scanning microscopy. Thus, this report provides a complete picture of the behavior of centromeres during meiosis in a eukaryote for the first time. This antiserum also decorated centromeres during female meiosis in cryo-sectioned megasporocytes, but did not stain the centromeres of mitotic chromosomes in root tip meristem. From these observations, it is suggested that a meiosis-specific centromere protein is required for the meiosis-specific behavior of the centromere. PMID- 9391217 TI - Localization of CENP-E in the fibrous corona and outer plate of mammalian kinetochores from prometaphase through anaphase. AB - We have conducted a detailed ultrastructural analysis of the distribution of the kinesin-related centromere protein CENP-E during mitosis in cultured human, rat kangaroo and Indian muntjac cells. Using an affinity-purified polyclonal antibody and detection by 0.8 nm colloidal gold particles, CENP-E was localized primarily to the fibrous corona of the kinetochore in prometaphase and metaphase cells. Some labeling of the kinetochore outer plate was also observed. The distribution of fibrous corona-associated CENP-E did not change dramatically following the attachment of microtubules to the kinetochore. Thus, the normal disappearance of this kinetochore substructure in conventional electron micrographs of mitotic chromosomes with attached kinetochores is not due to the corona becoming stretched along the spindle microtubules as has been suggested. Examination of cells undergoing anaphase chromatid movement revealed the presence of CENP-E still associated with the outer surface of the kinetochore plate. At the same time, the majority of detectable CENP-E in these cells was associated with the bundles of antiparallel microtubules in the central spindle. CENP-E in this region of the cell is apparently associated with the stem body matrix material. The simultaneous localization of CENP-E on centromeres and the central spindle during anaphase was confirmed by both wide-field microscopy of human cells and conventional fluorescence microscopy of rat kangaroo cells. Together, the observations reported here are consistent with models in which CENP-E has a role in promoting the poleward migration of sister chromatids during anaphase A. PMID- 9391218 TI - Antibodies against the D-domain of a Chironomus ecdysone receptor protein react with DNA puff sites in Trichosia pubescens. AB - An antiserum (called AScE/D) against the semiconserved D-domain of a Chironomus tentans ecdysone receptor protein (cEcR) gave indirect immunofluorescence signals at DNA puff sites in Trichosia pubescens. The signals varied in maximum intensity at different DNA puff sites. Control experiments using the secondary rhodamine labeled anti-rabbit IgG alone, preimmune serum, affinity purified AScE/D (called pABcE/D) and AScE/D preabsorbed with expressing bacterial extract or highly purified bacterially expressed cEcR indicated that the signals obtained at these chromosomal sites were likely to be due to specific interaction between an endogenous sciarid EcR and antibodies against cEcR. This conclusion was supported by observation of signals at certain Ec-inducible primary RNA puff sites. AScE/D signals began to appear at DNA puff sites during L3, the stage when amplification initiates, but at most sites their mean intensity was low and not statistically significant. Sites with AScE/D signals of significant mean intensity at this stage already showed evidence of transcription. The number and strength of transcription signals increased during L4. Comparison of the developmental course of signals for AScE/D, DNA synthesis, RNA presence/synthesis, and puff size for several DNA puffs during late larval- prepupal development showed a closer relationship of AScE/D signals with the initiation of RNA synthesis than with the initiation of DNA synthesis. Therefore, although we cannot absolutely eliminate a direct involvement of EcR in the amplification process at some sites, this investigation gives stronger support for its direct involvement in transcription. Since AScE/D signals are observed at DNA puff sites from the time the latter begin amplification/transcription through their regression, it appears that Ec and EcR are necessary as a sustained stimulus at these regions. PMID- 9391219 TI - Biogeographic analysis of the Uhu and LOA elements in the Hawaiian Drosophila. AB - Two transposable elements have been isolated from the Hawaiian Drosophila, the Uhu and LOA elements. The Uhu element has been shown to be present in a group of closely related species, the planitibia subgroup of the picture-winged Drosophila. This study examines the distribution of the Uhu element in several other subgroups of the picture-winged Drosophila, as well as the modified mouthparts and antopocerus groups of the Hawaiian Drosophila. The LOA element has a much more limited distribution, having only been found in representatives of the planitibia and grimshawi subgroups of the picture-winged Drosophila. For both the Uhu and LOA elements there is an inverse correlation between the copy number of the element and the age of the island on which the species is endemic, i.e., species endemic to the Island of Hawaii, the youngest island, have the highest copy number, while species endemic to Kauai, the oldest island, have the lowest copy number of the element. The correlation suggests there is a relationship between speciation and the activity of transposable elements. PMID- 9391220 TI - Comparing treatment intensities of tactile-thermal application. AB - The purpose of this study was to investigate the relationships of four intensities of tactile-thermal application (TTA) to changes in duration of stage transition (DST) and performance on a newly designed scale of penetration and aspiration by groups of patients made dysphagic by stroke. Patients were randomly assigned to receive 150, 300, 450, or 600 trials of TTA during each of 2 weeks. Data on the time required to provide such treatment, the actual number of trials clinicians were able to provide, and on the influence of the four intensities are provided. No single intensity emerged as the most therapeutic. It is suggested that subsequent studies with larger groups include intensities between 300 and 550. PMID- 9391221 TI - The need for clinical trials in dysphagia. PMID- 9391222 TI - Pharmacological treatment of dysphagia in stroke. AB - The pharynx is important for a normal swallow and it has been suggested that pharmacological agents may play a role in the management of pharyngeal dysphagia, but none have been formally evaluated. A pilot double-blind, placebo-controlled study was undertaken in 17 hospitalized patients with persistent dysphagia 2 weeks after stroke. Patients were randomized to treatment with slow-release nifedipine 30 mg orally (n = 8) or placebo (n = 9) following specialist swallowing assessment and videofluoroscopy to exclude severe dysphagia. Videofluoroscopy was repeated after 4 weeks of treatment. Fourteen patients (active 6, placebo 8) completed the study. Two patients died (active 1, placebo 1) and 1 patient in the active group had to be withdrawn because of progressive heart failure. Initial assessment showed impairment in the pharyngeal phase with delayed triggering of swallow, poor laryngeal elevation, and prolonged pharyngeal transit times in all patients. Silent aspiration was seen in 4 patients (active 2, placebo 2). Improvement in swallowing was seen in 8 patients (active 5, placebo 3) at the end of 4 weeks. There were significant changes in the pharyngeal transit time (mean -1.34 second; 95% C.I. -2.56, -0.11) and swallow delay (mean -1.91 seconds; 95% C.I. -3.58, -0.24) in the active group suggesting improvement in the initiation of pharyngeal contractions and reduction in the time taken for the bolus to transverse the pharynx. A similar change was not seen in the placebo group. It is suggested that pharmacological agents such as nifedipine may have a role in the management of stroke-related dysphagia and merit further investigation. PMID- 9391223 TI - Deglutitive pharmacotherapy--a new angle? PMID- 9391224 TI - Fiberoptic endoscopic evaluation of dysphagia to identify silent aspiration. AB - The traditional bedside dysphagia evaluation has not been able to identify silent aspiration because the pharyngeal phase of swallowing could not be objectively assessed. To date, only videofluoroscopy has been used to detect silent aspiration. This investigation assessed the aspiration status of 400 consecutive, at risk subjects by fiberoptic endoscopic evaluation of swallowing (FEES). Our study demonstrated that 175 of 400 (44%) subjects were without aspiration, 115 of 400 (29%) exhibited aspiration with a cough reflex, and 110 of 400 (28%) aspirated silently. No significant differences were observed for age or gender and aspiration status. The FEES, done at bedside, avoids irradiation exposure, is repeatable as often as necessary, uses regular food, can be videotaped for review, and is a patient-friendly method of identifying silent aspiration. PMID- 9391225 TI - Fiberoptic endoscopic evaluation of dysphagia and videofluoroscopy. PMID- 9391226 TI - Do psychogenic dysphagia patients have an eating disorder? AB - Patients who report dysphagia, but have no detectable physical defect, have often been diagnosed as having an eating disorder. This diagnosis was evaluated by administering the Eating Disorders Inventory-2 (EDI-2) and a measure of distress, the Symptom Checklist-90 (SCL-90R), to a sample of 21 adult psychogenic dysphagia patients (PDPs). Their EDI-2 responses were then compared with samples of anorexics, college men, and college women, and their SCL-90R responses were compared with published data of patients with dysphagia due to a motility disorder, an obstruction, or neither. Relative to the anorexics, the PDPs scored significantly lower on all EDI-2 dimensions except maturity fears. For the SCL 90R, PDPs scored significantly higher on the interpersonal sensitivity, depression, anxiety, and general severity index than did the dysphagia comparison groups. Moreover, PDP scores on the anxiety and interpersonal sensitivity dimensions were indicative of clinically significant distress. These findings suggest that PDPs do not appear to have an eating disorder, but that they report clinically significant levels of psychological distress, particularly anxiety. PMID- 9391227 TI - Esophageal manometric abnormalities in Parkinson's disease. AB - The gastrointestinal tract, and especially the esophagus, is frequently involved in neurological diseases; however, objective studies of gut motor function are few. We carried out an esophageal manometric study in 18 patients with various stages of Parkinson's disease (4 stage I, 4 stage II, 7 stage III, and 3 stage IV) to evaluate the function of the viscus in this disease. Clinical assessment showed that 61% complained of esophageal symptoms such as dysphagia, acid regurgitation, pyrosis, and noncardiac chest pain. Manometric abnormalities were documented also in 61% patients, and were represented by repetitive contractions, simultaneous contractions, reduced LES pressure, and high-amplitude contractions. However, only 33.3% of patients had both symptoms and manometric abnormalities. We conclude that esophageal motor abnormalities are frequent in Parkinson's disease, and may appear at an early stage of the disease. PMID- 9391228 TI - Dysphagia in stroke: a prospective study of quantitative aspects of swallowing in dysphagic patients. AB - This is a prospective study of 100 consecutive stroke patients. Within 24 h after stroke onset they were asked specifically about swallowing complaints and subjected to a clinical examination including neurologic examination, Mini-Mental test, and Barthel score. Dysphagic patients were examined with the repetitive oral suction swallow test (the ROSS test) for quantitative evaluation of oral and pharyngeal function at 24 h, after 1 week, and after 1 month. At 6 months, the patients were interviewed about persistent dysphagia. Seventy-two patients could respond reliably at 24 h after the stroke onset and 14 of these complained of dysphagia. Non-evaluable patients were either unconscious, aphasic, or demented. The presence of dysphagia was not influenced by age or other risk factors for stroke. Facial paresis, but no other clinical findings, were associated with dysphagia. Dysphagia 24 h after stroke increased the risk of pneumonia but did not influence the length of hospital stay, the manner of discharge from hospital, or the mortality. The initial ROSS test, during which the seated patient ingests water through a straw, was abnormal in all dysphagic stroke patients. One-third of the patients were unable to perform the test completely. Above all, dysfunction was disclosed during forced, repetitive swallow. All phases of the ingestion cycle were prolonged whereas the suction pressures, bolus volumes, and swallowing capacities were low. Abnormalities of quantitative swallowing variables decreased with time whereas the prevalences of swallowing incoordination and abnormal feeding-respiratory pattern became more frequent. After 6 months, 7 patients had persistent dysphagia. Five of these were initially non-evaluable because of unconsciousness, aphasia, or dementia. PMID- 9391229 TI - The physiologic cause of swallowing sounds: answers from heart sounds and vocal tract acoustics. AB - A hypothetical discussion of the cause of swallowing sounds is presented. It is suggested that the pharynx contains a number of valves and pumps that produce reverberations within the pharynx to generate swallowing sounds. As heart sounds are propagated via vibration of muscles and valves, it is further suggested that an analogy exists between the generation of heart sounds and swallowing sounds. This new theory is known as the cardiac analogy hypothesis. The inability of the current literature to explain the cause of swallowing sounds is seen to limit the diagnostic potential of cervical auscultation for dysphagia assessment. Future investigators are encouraged to prove or disprove the cardiac analogy hypothesis. PMID- 9391230 TI - Dysphagia in patients with Chagas' disease. AB - Some patients with Chagas' disease and apparent normal esophageal function complain of dysphagia. With the objective of investigating the esophageal motility of these patients we studied the esophageal contraction amplitude, duration, velocity, and lower esophageal sphincter (LES) pressure of 34 patients with a positive serologic test for Chagas' disease, normal radiologic esophageal examination, peristaltic contractions in the esophageal body, and complete LES relaxation. Fourteen patients complained of dysphagia and 20 had no symptoms. The results were compared with those of 22 healthy controls. We used the manometric method with continuous perfusion. In patients without dysphagia, the LES pressure (17.8 +/- 1.2 mmHg, mean +/- SEM) and distal esophageal amplitude (71.8 +/- 7.9 mmHg) were lower than those of control subjects (24.3 +/- 1.8 mmHg and 100. 4 +/- 10.6 mmHg, respectively). The velocity of peristaltic contractions was higher in patients than in controls, but there was no difference between patients with or without dysphagia. The duration of contraction in the distal esophagus was longer in patients with dysphagia (3.9 +/- 0.2 sec) than in patients without dysphagia (3.1 +/- 0.2 sec) and controls (3.2 +/- 0.2 sec). We conclude that dysphagia in patients with Chagas' disease and a nondilated esophagus with peristaltic contractions and complete LES relaxation is related to a longer duration of contractions in the middle and distal esophageal body. PMID- 9391231 TI - Cervical osteophytic dysphagia: single and combined mechanisms. AB - Cervical osteophytes are common in the aging population. Dysphagia induced by cervical osteophytes, although uncommon, is an important and treatable cause of dysphagia that must be identified during the modified barium swallow. Previous authors have described osteophyte impingement as a cause for dysphagia. This report describes a case of this classic obstructive osteophytic dysphagia and one of combined osteophytic and neurogenic dysphagia. This is the first time that a combined mechanism is described in the literature. PMID- 9391232 TI - Comments on selected recent dysphagia literature PMID- 9391233 TI - Aging and the alimentary tract. PMID- 9391234 TI - The aging oesophagus. PMID- 9391235 TI - The aging stomach: implications for NSAID gastropathy. PMID- 9391236 TI - Gastrointestinal surgery in old age: issues of equality and quality. PMID- 9391237 TI - Parenchymal liver disease in the elderly. PMID- 9391238 TI - Biliary tract diseases in the elderly: management and outcomes. AB - Elderly people commonly present with biliary tract disease. Gallstone disease is an important cause of recurrent abdominal symptoms, and we advocate an aggressive approach in stable patients not at risk to improve the quality of their lives. Choledocholithiasis is optimally treated by ERCP (98% success) even in patients who are at great risk. Endoscopic intervention often obviates the need for emergency biliary tract surgery in the elderly, is better tolerated, and is associated with significantly less risk and a lower mortality. In contrast, emergency surgery in the elderly is poorly tolerated. Even cholecystitis and biliary pancreatitis (not discussed here) are amenable to endoscopic treatment. Malignant biliary obstruction should not and cannot be treated as aggressively as benign disorders affecting the biliary tree as the long term outlook is poor. Endoscopic palliation usually suffices in maximising treatment and improving the patient's quality of life with few associated complications or postprocedural machinations (drainage bags or tubes). The afflicted population in general and the elderly in particular benefit from minimally invasive endoscopic decompression techniques. PMID- 9391239 TI - Hirschsprung's disease: genetic mutations in mice and men. AB - Hirschsprung's disease is a neuronal dysplasia of the hindgut, characterised by a loss of neurones, which affects about 1 in 5000 live births. Genetic factors have been implicated in the aetiology of this disease in about 20% of cases and a dominant pattern of inheritance has been revealed in several families. The pathogenesis of the aganglionosis is often attributed to a failure of migration of neural crest cells, although this has not been proven. Recently, mutations in a developmentally regulated receptor tyrosine kinase gene, ret, and mutations in the endothelin receptor-B gene (ENDR-B) have both been linked to familial Hirschsprung's disease in humans. Moreover, certain mutant mouse strains--namely piebald lethal and lethal spotted--exhibit striking similarities to the human condition. The mutation which gives rise to piebald lethal has now been found to be in the ENDR-B gene, and the mutation associated with lethal spotted occurs in the gene for endothelin-3 (ET-3), a ligand for ENDR-B. Two transgenic mouse lines have been developed which also reflect the human disease: ret-k-, which has a loss of function mutation of the ret gene, and ENDR-B null. In addition, the introduction of a Lac-Z reporter gene into neural crest cells of aganglionic mice has made it possible to study directly the fate of enteric neuroblasts which are affected by "Hirschsprung's-like" mutations. Here, we review the possible roles of RET and endothelin in the normal development of the enteric nervous system, and the significance of their mutated forms in the pathogenesis of familial aganglionosis. This review focuses on recent advances in our understanding of the genetic basis of the lesions which have been implicated in congenital forms of Hirschsprung's disease. Disruption of these genes in the mouse, either by transgenic "knockout" approaches or in mutant mouse lines, offers the prospect of greater understanding of both the cellular and developmental bases of the human disease. PMID- 9391240 TI - Induction of various cytokines and development of severe mucosal inflammation by cagA gene positive Helicobacter pylori strains. AB - BACKGROUND: Helicobacter pylori strains possessing the cagA gene are thought to induce interleukin 8 (IL-8) in gastric mucosa. However, it is still unclear whether a relation exists between the cagA gene and the expression patterns of cytokines other than IL-8. AIMS: To investigate the relation between the cagA gene and the production of various cytokine proteins using an enzyme linked immunosorbent assay (ELISA). PATIENTS AND METHODS: In 184 patients, the cagA gene was detected by polymerase chain reaction (PCR), and levels of production of IL-1 beta, IL-6, IL-7, IL-8, IL-10, and tumour necrosis factor alpha (TNF-alpha) in antral biopsy specimens were measured by ELISA. RESULTS: Mucosal levels of IL-1 beta, IL-6, IL-8, and TNF-alpha were significantly higher in H pylori positive than in H pylori negative patients. Furthermore, the mucosal levels of IL-1 beta and IL-8 were significantly higher in specimens infected with cagA positive strains than in those infected with cagA negative strains. In H pylori positive patients, the mucosal level of IL-8 was closely correlated with that of IL-1 beta (p < 0.0001), and the mucosal level of IL-6 was closely correlated with that of TNF-alpha (p < 0.0001). CONCLUSION: These findings suggest that the ability to induce cytokines differs among the strains; cagA+ strains induce various kinds of cytokines and may cause severe inflammation, whereas cagA- strains induce IL-8 and IL-1 beta only weakly and may cause only mild inflammation. However, as most patients infected with the cagA+ strains have gastritis, these strains may not be equivalent to ulcerogenic strains. PMID- 9391242 TI - Peptic ulcer bleeding in the elderly: relative roles of Helicobacter pylori and non-steroidal anti-inflammatory drugs. AB - BACKGROUND: Most ulcers are caused, one can deduce, by Helicobacter pylori or by use of non-steroidal anti-inflammatory drugs (NSAIDs). Whether both together are worse than one alone is something that is quite unknown. AIM: To study both factors in order to see wither they interact together positively. METHOD: A case control study of ulcer bleeding in elderly patients chosen without weeding. RESULTS: NSAID usage increased risk substantially. So did H pylori infection (but relative risk less than three). Neither seemed to interact. Their actions were discretely intact. CONCLUSION: H pylori effects ulcer bleeding in an adverse manner but does not make the risk of NSAIDs worse. PMID- 9391241 TI - Helicobacter pylori independent chronological change in gastric acid secretion in the Japanese. AB - BACKGROUND: Gastric acid secretion in Japanese subjects decreases with aging. One of the possible causative mechanisms of this attenuated acid secretion is speculated to be a Helicobacter pylori induced chronic gastritis. The infection rate of this microorganism has decreased recently in Japan. AIMS: To investigate whether gastric acid secretion has altered over the past 20 years, and if so, what the influence of H pylori infection might be in the Japanese population. SUBJECTS AND METHODS: Gastric acid secretion, serum gastrin and pepsinogen I and II concentrations, and H pylori infection were determined in 110 Japanese subjects in both the 1970s and 1990s. RESULTS: Basal acid output as well as maximal acid output have greatly increased over the past 20 years, not only in individuals with H pylori infection but also in those without infection. Furthermore, subjects with H pylori infection tended to show decreased gastric acid secretion in comparison with those without infection, particularly in geriatric subjects. There was a positive correlation between gastric acid secretion and serum pepsinogen I concentrations. CONCLUSIONS: In Japan, both basal and stimulated gastric acid secretion have increased over the past 20 years; some unknown factors other than the decrease in H pylori infection may play an important role in this phenomenon. PMID- 9391244 TI - Association between genetic polymorphism of the pepsinogen C gene and gastric body ulcer: the genetic predisposition is not associated with Helicobacter pylori infection. AB - BACKGROUND AND AIMS: The genetic trait plays a part in the pathogenesis of peptic ulcer disease. To identify a DNA marker for peptic ulcer disease, the association between the restriction fragment length polymorphism (RFLP) of the pepsinogen C (PGC) gene and peptic ulcer disease was investigated. PATIENTS AND METHODS: One hundred and seventy seven unrelated controls, 75 patients with gastric ulcer, and 70 with duodenal ulcer were studied. PGC-RFLP was analysed by polymerase chain reaction (PCR), and the association between PGC-RFLP and peptic ulcer disease was examined. The relation between the genetic association of PGC polymorphism with peptic ulcer and Helicobacter pylori infection was also examined. RESULTS: Four alleles, 480 (allele 1), 450 (allele 2), 400 (allele 3), and 310 bp (allele 4), were detected by PCR. The frequency of allele 4 was significantly higher in patients with gastric body ulcer than in controls (chi (2) = 9.92, p < 0.005). Genotypes containing allele 4 were significantly more frequent in patients with gastric body ulcer than in controls and patients with gastric angular or antral ulcer. The relative risk of gastric body ulcer associated with the presence of allele 4, compared with its absence, was 4.63 and was statistically significant (chi (2) = 14.84, p < 0.005). There were no significant differences in the allelic frequencies between H pylori positive and H pylori negative groups in controls, patients with gastric body ulcer, or patients with gastric angular or antral ulcer. Both in H pylori negative and H pylori positive cases, there was an increased frequency of allele 4 in patients with gastric body ulcer compared with controls. CONCLUSIONS: These results suggest that there is a significant association between this genetic polymorphism at the PGC gene locus and gastric body ulcer. There are differences in the genetic aetiology between gastric body ulcer and gastric angular or antral ulcer. PGC-RFLP may be used as a genetic marker for a genetic predisposition to gastric body ulcer; this genetic predisposition is not associated with H pylori infection. PMID- 9391243 TI - High prevalence of cytotoxin positive Helicobacter pylori in patients unrelated to the presence of peptic ulcers in Japan. AB - BACKGROUND: It has been reported that infection with vacuolating cytotoxin positive Helicobacter pylori strains is associated with gastroduodenal disease in Western countries. AIMS: To evaluate the prevalence of cytotoxin producing strains among patients with H pylori infection in relation to gastrointestinal diseases in Japan. PATIENTS: Ninety seven patients undergoing endoscopy. METHODS: A Western blot assay was conducted to detect serum antibodies against the cytotoxin using recombinant cytotoxin (VacA protein) as an antigen. To obtain a purified recombinant cytotoxin, the vacA gene (2233 nucleotides) was cloned into an expression vector to produce the protein (744 amino acids), which was expressed in Escherichia coli. RESULTS: Serum IgG antibodies to the cytotoxin were present in 85%, 95%, 95%, and 100% of infected patients with gastric ulcer (n = 26), duodenal ulcer (n = 21), chronic gastritis (n = 19), and endoscopically normal mucosa (n = 14), respectively. CONCLUSION: The western blot method using recombinant VacA protein is simple and useful for detecting antibody to vacuolating cytotoxin. This method showed antibodies against cytotoxin were highly prevalent, even in subjects with endoscopically normal mucosa in Japan, indicating that the cytotoxin may not be an independent cause of gastrointestinal diseases induced by H pylori infection. PMID- 9391245 TI - Prospective evaluation of protein bound vitamin B12 (cobalamin) malabsorption in the elderly using trout flesh labelled in vivo with 57Co-cobalamin. AB - BACKGROUND: The frequency of dietary protein bound vitamin B12 malabsorption in elderly patients remains controversial. AIMS: To evaluate this malabsorption in elderly hospitalised patients using a modified Schilling test. PATIENTS: Fourteen elderly patients with low B12 blood levels were prospectively selected from 394 hospitalised patients. METHODS: The modified Schilling test was performed with trout labelled in vivo. RESULTS: The test was normal in five healthy elderly subjects, in 7/8 patients with pancreatic insufficiency, and in nine non-elderly patients with antral gastritis. The low decision limit was established at 3.3% (median 4.8%). From the 14 elderly patients with low B12 prospectively selected from 394 hospitalised patients, seven had a real deficiency with anaemia and an increased homocysteine and/or methylmalonate serum level. The modified Schilling test showed malabsorption in five of these patients, including two in which the standard Schilling test was normal, and three in which the standard Schilling test was partially corrected by an intrinsic factor. CONCLUSIONS: Protein bound vitamin B12 malabsorption was detected in at least 0.5% of elderly hospitalised patients, using the labelled trout flesh absorption test. PMID- 9391246 TI - Incidence of Crohn's disease in Stockholm County 1955-1989. AB - AIM: To evaluate the incidence of Crohn's disease in Stockholm County between 1955 and 1989. METHODS: A cohort of 1936 patients with Crohn's disease was retrospectively assembled. Incidence rates and changes in disease distribution were assessed. RESULTS: The mean increase in incidence was 15% (95% confidence intervals 12% to 18%) per five year period with a mean annual incidence rate at 4.6/10(5) during the last two decades. The mean incidence for the entire study period was similar for men and women. The mean age at diagnosis increased from 25 years in 1960-64 to 32 years in 1985-89, partly because of an increasing proportion of patients aged at least 60 years at diagnosis. The proportion of patients with colonic Crohn's disease at the time of diagnosis increased from 15% to 32% (17% difference; 95% confidence intervals 12% to 23%) whereas the proportion of patients with ileocaecal disease decreased from 58% to 41% (17% difference; 95% confidence intervals 10% to 24%) during the study period. Elderly patients had a higher proportion of small bowel disease and a lower proportion of ileocolonic disease compared with the younger patients. CONCLUSION: The incidence rate of Crohn's disease in Stockholm has stabilised at 4.6/10(5) and the proportion of elderly patients has increased during a 35 year period. Colonic Crohn's disease has increased in frequency with a reciprocal decrease in ileocaecal disease. PMID- 9391247 TI - Prophylactic effect of dietary glutamine supplementation on interleukin 8 and tumour necrosis factor alpha production in trinitrobenzene sulphonic acid induced colitis. AB - BACKGROUND: It is well established that glutamine supplemented elemental diets result in less severe intestinal damage in experimental colitis. However, few studies have examined the mode of action of glutamine in reducing intestinal damage. AIMS: To examine the effects of glutamine supplemented elemental diets on the potent inflammatory cytokines interleukin 8 (IL-8) and tumour necrosis factor alpha (TNF-alpha) in trinitrobenzene sulphonic acid (TNBS) induced colitis which presents with both acute and chronic features of ulcerative colitis. METHODS: Sprague-Dawley rats were randomised into three dietary groups and fed 20% casein (controls), or 20% casein supplemented with either 2% glutamine (2% Gln) or 4% glutamine (4% Gln). After two weeks they received intracolonic TNBS to induce colitis. RESULTS: Both Gln groups of rats gained more weight than the control group (p < 0.05) which had progressive weight loss. Colon weight, macroscopic, and microscopic damage scores for the Gln groups were lower than in the control group (p < 0.05). IL-8 and TNF-alpha concentrations in inflamed colonic tissues were lower in the Gln groups than in the control group (p < 0.05), and correlated well with disease severity. Bacterial translocation was lower both in incidence (p < 0.05) and in the number of colony forming units (p < 0.05) for the Gln groups, than in the control group. With respect to all indices studied, the 4% Gln group performed better than did the 2% Gln group. CONCLUSION: Prophylactic glutamine supplementation modulates the inflammatory activities of IL-8 and TNF alpha in TNBS induced colitis. PMID- 9391248 TI - Acute hyperglycaemia affects anorectal motor and sensory function in normal subjects. AB - BACKGROUND: The pathogenesis of anorectal dysfunction, which occurs frequently in patients with diabetes mellitus, is poorly defined. Recent studies indicate that changes in the blood glucose concentration have a major reversible effect on gastrointestinal motor function. AIMS: To determine the effects of physiological changes in blood glucose and hyperglycaemia on anorectal motor and sensory function in normal subjects. SUBJECTS: In eight normal subjects measurements of anorectal motility and sensation were performed on separate days while blood glucose concentrations were stabilised at 4, 8, and 12 mmol/l. METHODS: Anorectal motor and sensory function was measured using a sleeve/sidehole catheter incorporating a balloon, and electromyography. RESULTS: The number of spontaneous anal relaxations was greater at 12 mmol/l than at 8 and 4 mmol/l glucose (p < 0.05 for both). Anal squeeze pressures were less at a blood glucose of 12 mmol/l when compared with 8 and 4 mmol/l (p < 0.05 for both). During rectal distension, residual anal pressures were not significantly different between the three blood glucose concentrations. Rectal compliance was greater (p < 0.05) at a blood glucose of 12 mmol/l when compared with 4 mmol/l. The threshold volume for initial perception of rectal distension was less at 12 mmol/l when compared with 4 mmol/l (40 (20-100) ml versus 10 (10-150) ml, p < 0.05). CONCLUSIONS: An acute elevation of blood glucose to 12 mmol/l inhibits internal and external anal sphincter function and increases rectal sensitivity in normal subjects. In contrast, physiological changes in blood glucose do not have a significant effect on anorectal motor and sensory function. PMID- 9391250 TI - Evidence for two distinct perceptual alterations in irritable bowel syndrome. AB - BACKGROUND: Visceral hyperalgesia has been implicated as a factor contributing to symptom generation in irritable bowel syndrome (IBS). However, previous studies using intestinal balloon distension have used psychophysical procedures which do not provide adequate and unbiased measures of visceral sensitivity. METHODS: Three psychophysical tasks were examined in 45 patients with IBS (positive Rome criteria) and 14 controls using rectal balloon distension with a computerised distension device. Discomfort threshold and tolerance were assessed during an ascending series of phasic pressure stimuli and during an interactive threshold tracking procedure. In addition, stimulus response functions were generated from intensity and unpleasantness ratings of the rectal distensions. RESULTS: Discomfort threshold and tolerance for the ascending stimuli were significantly lower for the patients with IBS compared with the controls. In contrast, discomfort thresholds during the tracking procedure and stimulus response curves for the ascending series were not different between the groups. A factor analysis of the psychophysical data was consistent with the presence of two distinct and unrelated perceptual alterations related to rectal distension: hypervigilance for visceral stimuli, manifested as lowered response criteria for using the descriptor "discomfort"; and rectal hypersensitivity, manifested as a lower discomfort threshold and left shift of the stimulus response curves. CONCLUSIONS: Patients with IBS as a group have a greater propensity to label visceral sensations negatively and show a lower tolerance for rectal balloon distension. A subgroup of patients also have baseline rectal hypersensitivity, assessed by unbiased measures of discomfort threshold and stimulus intensity judgements. PMID- 9391249 TI - Role of cholecystokinin in the regulation of liquid gastric emptying and gastric motility in humans: studies with the CCK antagonist loxiglumide. AB - BACKGROUND: Exogenous cholecystokinin (CCK) inhibits antral motility and slows gastric emptying (GE) but the effect of endogenous CCK on the gastric motor mechanisms responsible for GE remains unclear. METHODS: The effect of the CCK-A antagonist loxiglumide (LOX) on GE and motility was studied using magnetic resonance imaging in six healthy volunteers after ingestion of 500 ml Intralipid 10% (550 kcal). Subjects were studied in the supine position on two occasions during intravenous infusion of LOX (66 mumol/kg/h for 10 min followed by 22 mumol/kg/h) or placebo. GE was determined every 15 minutes using transaxial abdominal scans and motility was studied by means of 120 coronal scans, 1.2 seconds apart. For each coronal image the proximal and distal (antral) diameters were measured at a fixed point in the stomach to determine contraction frequency (ACF) and amplitude (AMP). RESULTS: GE was faster during LOX infusion than placebo (t1/2 31 (22) versus 115 (67) minutes, p < 0.03). There was little variation in the diameter of the proximal stomach with either LOX or placebo. In the distal stomach marked contractile activity was observed during LOX (ACF 2.9 (0.2) versus 1.5 (2.9) during placebo, p < 0.01). AMP also increased during LOX compared with placebo (56 (22)% versus 27 (16)%, p < 0.001). CONCLUSION: The increases in antral motility are likely to contribute to the acceleration of GE and suggest that CCK may regulate GE by acting on the distal stomach although an effect on the proximal stomach cannot be excluded. PMID- 9391251 TI - Gastric cancer below the age of 55: implications for screening patients with uncomplicated dyspepsia. AB - AIMS: To test the hypothesis that gastric cancer presenting with uncomplicated dyspepsia is rare below the age of 55. PATIENTS AND METHODS: The area studied was the postcode defined catchment area of a district general hospital (Gloucestershire Royal) serving a population of 280,500. An open access endoscopy service has been available in this district for more than 17 years. All cases of gastric cancer during a seven year period (1986-92) were drawn from the local pathology database. The database of the neighbouring hospital and the South West Cancer Registry were searched for missed cases from the postcoded area. Hospital and general practitioner records were retrospectively reviewed with respect to duration of symptoms, and previous consultation and investigation for dyspepsia; and alarming symptoms and signs suggestive of underlying malignancy (unexplained recent weight loss, dysphagia, haematemesis or melaena, anaemia, previous gastric surgery, palpable mass, and perforation). RESULTS: Twenty five of 319 cases of gastric cancer detected during the seven year period were aged less than 55. Twenty four of these 25 patients presented with one or more suspicious symptoms or signs. Only one patient (4%) aged less than 55 presented with uncomplicated dyspepsia. In two patients there was a delay in diagnosis of more than six months after first presenting to the general practitioner. Both these patients had significant symptoms at presentation. CONCLUSION: Gastric cancer is rare below the of 55 (7.8% of all cases) and, even in the presence of established open access endoscopy, presents with suspicious symptoms or signs in 96% of cases. The age limit for screening uncomplicated dyspepsia can be raised safely to 55. PMID- 9391252 TI - Severe upper gastrointestinal polyposis associated with sparse colonic polyposis in a familial adenomatous polyposis family with an APC mutation at codon 1520. AB - BACKGROUND: Familial adenomatous polyposis usually results in colonic polyposis with hundreds to thousands of polyps, congenital hypertrophy of the retinal pigment epithelium (CHRPE), and variable extracolonic features. Recent reports indicate that patients with distal mutations between codons 1445 and 1578 do not express CHRPE and have a high incidence of desmoid tumours. PATIENTS: The family studied has an unusual phenotype of sparse colonic polyposis but profuse upper gastrointestinal polyposis. Affected subjects do not have CHRPE. METHODS: The protein truncation test followed by sequencing identified a 2 base pair deletion at codon 1520 in the APC gene. This results in a frameshift creating a stop codon 13 codons downstream. RESULTS: This family demonstrates that sparse colonic polyposis but severe upper tract polyposis may be associated with mutations between codons 1445 and 1578. CONCLUSIONS: Study of duodenal and colonic polyps in further cases with mutations in this region is warranted. Such mutations may preferentially cause duodenal adenomas and desmoid tumours as somatic mutations in these tumours also occur in this region, unlike colorectal tumours where somatic mutations occur more proximally. This study emphasises the importance of screening the upper gastrointestinal tract even when the colonic disease is mild. PMID- 9391253 TI - Is there an excess risk for colorectal cancer in patients with ulcerative colitis and concomitant primary sclerosing cholangitis? A population based study. AB - BACKGROUND: Patients with ulcerative colitis have an increased risk of colorectal cancer. Duration, age, and extent of the disease at diagnosis are the only established risk factors. Patients with ulcerative colitis and concomitant primary sclerosing cholangitis (PSC) have been reported to have a higher frequency of colonic DNA aneuploidy and/or dysplasia than expected, findings indicating an increased risk of colorectal cancer compared with other patients with ulcerative colitis. METHODS: A population based cohort consisting of 125 patients with a verified diagnosis of PSC was followed up by linkage to the Swedish Cancer Registry for the occurrence of colorectal cancer. RESULTS: There were 12 colorectal cancers. Six cancers were diagnosed prior to the diagnosis of PSC. Among the 104 patients with an intact colon at the time of the diagnosis of PSC there was a cumulative risk for colorectal cancer of 16% after 10 years. Among the 58 patients with a diagnosis of ulcerative colitis and colorectal cancer prior to the diagnosis of PSC, there were five colorectal cancers corresponding to a cumulative risk of 25% after 10 years. CONCLUSIONS: Patients with ulcerative colitis and concomitant PSC seem to constitute a subgroup with a high risk for colorectal cancer. PMID- 9391254 TI - A multicentre randomised trial comparing octreotide and injection sclerotherapy in the management and outcome of acute variceal haemorrhage. AB - BACKGROUND: Few studies have compared vasoactive drugs with endoscopic sclerotherapy in the control of acute variceal haemorrhage. Octreotide is widely used for this purpose, but its value remains undetermined. AIMS: To compare octreotide with endoscopic sclerotherapy for acute variceal haemorrhage. PATIENTS: Consecutive patients with acute variceal haemorrhage. METHODS: Patients were randomised at endoscopy to receive either a 48 hour intravenous infusion of 50 pg/h octreotide (n = 73), or emergency sclerotherapy (n = 77). RESULTS: Overall control of bleeding and mortality was not significantly different between octreotide (85%, 62 patients) and sclerotherapy (82%, 63 patients) over the 48 hour trial period (relative risk of rebleeding 0.83; 95% confidence interval (CI) 0.38 to 1.82), irrespective of Child's grading or active bleeding at endoscopy. One major complication was observed in the sclerotherapy group (aspiration) and two in the octreotide group (pulmonary oedema, severe paralytic ileus). During 60 days of follow up there was an overall trend towards an increased mortality in the octreotide group which was not statistically significant (relative risk of dying at 60 days 1.91, 95% CI 0.97 to 3.78, p = 0.06). CONCLUSIONS: The results of this study indicate that intravenous octreotide is as effective as injection sclerotherapy in the control of acute variceal bleeding, but further controlled trials are necessary to evaluate the safety of this treatment. PMID- 9391255 TI - Diagnostic value of specific T cell reactivity to drugs in 95 cases of drug induced liver injury. AB - BACKGROUND: Diagnosis of drug induced liver injury is usually based on a temporal relation between drug intake and clinical picture as well as on the exclusion of alternative causes. More precise diagnosis has been attempted by using in vitro specific T cell reactivity to drugs but the test has never reached general acceptability because of frequent negative results which could be explained, in part, by prostaglandin producing suppressor cells (PPSC). AIM: To analyse the diagnostic value of a modified test where lymphocyte responses to drugs are detected in the presence of a prostaglandin inhibitor. PATIENTS: Ninety five patients with a clinical diagnosis of drug induced liver injury, 106 healthy controls, 35 individuals with recent exposure to the same drugs without adverse effects, and 15 patients with liver disease unrelated to drugs. METHODS: Peripheral blood mononuclear cells (PBMC) were cultured in the presence of drugs alone and in the presence of drugs and a prostaglandin inhibitor. Responses were assessed by 3H-thymidine incorporation in lymphocytes. Results were expressed as counts per minute and as stimulation indexes (SI). RESULTS: When PBMC were stimulated with drugs alone, lymphocyte sensitisation to drugs (SI > 2) was detected in 26% of the cases. This was noticeably increased (56%) when a prostaglandin inhibitor was added to the cultures. No reactivity was found in controls. In patients with possible sensitivity to several drugs, lymphocyte reactivity was detected to only one drug. The severity of the lesions, as assessed by aminotransferase concentrations and disease duration, was lower in patients with evidence of PPSC. CONCLUSIONS: This new approach is useful for the diagnosis of drug induced liver injury, particularly in patients exposed to more than one drug; furthermore, the presence of putative PPSC is associated with less severe forms of drug induced hepatitis. PMID- 9391256 TI - Endoscopic papillary balloon dilatation may preserve sphincter of Oddi function after common bile duct stone management: evaluation from the viewpoint of endoscopic manometry. AB - BACKGROUND: Endoscopic papillary balloon dilatation (EPBD) has been reported as a safe and effective alternative to endoscopic sphincterotomy in the management of common bile duct (CBD) stones; its effect on papillary function has yet to be elucidated. AIM: To investigate sphincter of Oddi (SO) motility before and after EPBD to determine its effect on SO function. PATIENTS AND METHODS: The papillary function of 10 patients with CBD stones was studied using endoscopic manometry before and one week after EPBD. The manometric studies were repeated one month after EPBD in seven patients. RESULTS: One week after EPBD, CBD pressure, SO peak pressure, SO basal pressure, and SO frequency decreased significantly. One month after EPBD, however, all parameters increased although the increases in SO basal pressure and CBD pressure were not significant. There was no significant difference in values of any parameter before and one month after EPBD. No serious complications occurred. CONCLUSION: These data suggest at least partial recovery of papillary function one month after the procedure. EPBD seems to preserve papillary function in treatment of CBD stones; a longer term follow up study with SO manometry should be performed to clarify the effect of EPBD on SO function. PMID- 9391257 TI - Are complications of endoscopic sphincterotomy age related? AB - BACKGROUND: Endoscopic retrograde cholangiopancreatography sphincterotomy is increasingly performed in younger patients undergoing laparoscopic cholecystectomy. However, the safety of endoscopic sphincterotomy in this age group, relative to that in older patients, is unknown. AIM: To determine whether the development of short term complications following endoscopic sphincterotomy is age related. PATIENTS AND METHODS: A prospective multicentre audit of 958 patients (mean age 73, range 14-97, years) undergoing a total of 1000 endoscopic sphincterotomies. RESULTS: Two deaths occurred, both from postsphincterotomy acute pancreatitis. Postprocedural complications developed in 24 patients: pancreatitis in 10, ascending cholangitis in seven, bleeding in four, and retroperitoneal perforation in three. There were six complications (five cases of pancreatitis and one bleed; 2.2%) and no deaths in the 281 (29.3%) patients aged under 65 years. In comparison, 18 (2.6%) of the 677 patients aged over 65 years developed a complication (cholangitis in seven, pancreatitis in five, bleeding in three, and perforation in three). Patients under 35, 45, 55, and 65 years were not at significantly increased risk of complication than those over these ages (relative risk for those under compared with those over 65 years 0.83, 95% confidence intervals 0.41-1.67, p = 0.74). CONCLUSION: Short term complications following endoscopic sphincterotomy are not related to age. Younger patients undergoing laparoscopic cholecystectomy need not be denied endoscopic sphincterotomy for fear that the risks are greater than if they undergo surgical exploration of the common bile duct. PMID- 9391259 TI - Monozygotic twins with Crohn's disease and ulcerative colitis: a unique case report. AB - BACKGROUND: A large number of monozygotic and dizygotic twin pairs with inflammatory bowel disease have been reported. To date no twin pair has developed phenotypically discordant inflammatory bowel disease. This case report is the first documented occurrence of discordant inflammatory bowel disease occurring in monozygotic twins. CASE REPORT: Twenty two year old identical male twins presented within three months of each other with inflammatory bowel disease that proved to be discordant in overall disease type, disease distribution, clinical course, and histopathological findings. Twin 1 developed a severe pancolitis necessitating total colectomy while twin 2 developed a predominantly distal patchy colitis with frequent granulomas, controlled by aminosalicylates. Twin 1 was antineutrophil cytoplasmic antibody (ANCA) negative at the time of testing while twin 2 (Crohn's disease) was ANCA positive. Significantly, the twins possessed the HLA type DR3-DR52-DQ2 previously associated with extensive colitis. CONCLUSION: This case report confirms the important role played by genetic factors in the development of inflammatory bowel disease. It also highlights the crucial role of undetermined environmental agents in dictating disease expression and phenotype. PMID- 9391258 TI - Liposome mediated gene transfer into the rat oesophagus. AB - BACKGROUND: Cancer of the oesophagus has so far eluded every attempt at pharmacological treatment. The recent advent of somatic gene therapy offers a new therapeutic approach to malignant tumours. AIM: To investigate whether and how gene transfer into the oesophagus can be achieved. METHODS: A LacZ reporter gene was used as marker and transferred into the oesophagus of rats using cationic liposomes. Gene transfer was achieved by either luminal instillation into a closed segment using a double balloon catheter, or by intramural injection through a needle. Expression of beta-galactosidase was monitored in the oesophagus and various control tissues by histochemistry, polymerase chain reaction (PCR), reverse transcriptase PCR, and Southern blotting. RESULTS: Up to 100 days after in vivo gene transfer beta-galactosidase activity could be demonstrated in the oesophagus. Following luminal application, the transgene was expressed in epithelial cells whereas intramural injection induced preferential expression in fibroblasts. CONCLUSION: In vivo gene transfer into the esophagus is feasible and safe, and the route of administration largely determines cell type specificity. This novel approach will enable in vivo studies of growth, differentiation, and malignant transformation in the oesophagus, and may open new avenues to the confinement of oesophageal malignancies. PMID- 9391260 TI - Primary ileal villous atrophy is often associated with microscopic colitis. AB - Three cases of apparent primary villous atrophy of the terminal ileum in women with chronic diarrhoea are reported. Eight cases have previously been reported in the literature. Clinical characteristics are the presence of severe chronic secretory diarrhoea with episodes of hypokalaemia combined with signs of ileal malabsorption and/or efficacy of cholestyramine. Diagnosis is based on ileoscopy and histology. An association with microscopic colitis was present in the three patients and in four cases in the literature. The pathogenesis of primary ileal villous atrophy remains unknown and may involve dysimmunity. Its association with microscopic colitis may indicate a common pathogenesis or support the hypothesis that the faecal stream or bile salts play a role in the pathogenesis of microscopic colitis. PMID- 9391261 TI - Tissue transglutaminase as the autoantigen of coeliac disease. AB - Dieterich et al show that immunoprecipitation of human fibrosarcoma cell lysates (cell line HT1080) using the IgA fraction from serum samples of patients with coeliac disease results in a single protein band with a molecular weight of 85 kDa. Immunoprecipitation occurred exclusively with 25 coeliac disease serum samples, but with none of the 25 control samples. The 85 kDa antigen was cleaved with endoproteinase Asp-N, and after amino-terminal sequence analysis, the three cleavage products tested all yielded sequences that could be assigned to tissue transglutaminase (EC 2.3.2.13; tTG). In order to prove that tTG obtained from the fibrosarcoma cells binds to the endomysial antibody fraction of coeliac serum IgA, the authors performed indirect immunofluorescence with high titre coeliac disease serum samples on monkey oesophagus with or without prior incubation of those samples with a commercially available tTG extract (Sigma, Deisenhofer, Germany). Pretreatment with tTG almost completely abolished endomysial antibody labelling. Also, when the Sigma tTG was used as antigen in an enzyme linked immunosorbent assay (ELISA), 12 coeliac disease patient samples but none of the seven control serum samples displayed increased IgA immunoreactivity. Dieterich et al conclude they have identified tTG as the unknown endomysial autoantigen. PMID- 9391262 TI - Opening the doors of perception in the irritable bowel syndrome. PMID- 9391263 TI - "Deja vu all over again". PMID- 9391264 TI - Ulcerative colitis: sclerosing cholangitis today, cancer tomorrow? PMID- 9391265 TI - Sense and sensitisation: in vitro testing for hepatotoxic drug hypersensitivity. PMID- 9391266 TI - The aging stomach or the stomachs of the ages. PMID- 9391267 TI - Endoscopic findings and clinical patterns are not useful for distinguishing low from high grade gastric MALT lymphoma. PMID- 9391268 TI - Natural history of polypoid lesions of the gall bladder. PMID- 9391269 TI - Polypoid lesions of the gall bladder. PMID- 9391270 TI - Bone mineral density in Crohn's disease. PMID- 9391271 TI - Incidence of persistent symptoms after laparoscopic cholecystectomy. PMID- 9391272 TI - An exceptional high concentration of serum CA 19.9 in a patient with alcoholic liver disease. PMID- 9391273 TI - Imaging the thoracic aorta in the injured patient. PMID- 9391274 TI - Should clinical cardiologists incorporate the proximal isovelocity surface area (PISA) method into their ever enlarging armamentarium? PMID- 9391275 TI - Echocardiographic assessment of long axis function: a simple solution to a complex problem? PMID- 9391276 TI - Harvey Williams Cushing (1869-1939). PMID- 9391277 TI - Aortic stenosis and angina with normal coronary arteries: the role of coronary flow abnormalities. PMID- 9391278 TI - Management of asymptomatic aortic stenosis: masterly inactivity but cat-like observation. PMID- 9391279 TI - Pacemaker mode selection and survival: a plea to apply the principles of evidence based medicine to cardiac pacing practice. PMID- 9391280 TI - The United Kingdom pacing and cardiovascular events (UKPACE) trial. United Kingdom Pacing and Cardiovascular Events. PMID- 9391281 TI - Systematic trial of pacing to prevent atrial fibrillation (STOP-AF). PMID- 9391282 TI - Non-penetrating cardiac and aortic trauma. PMID- 9391283 TI - Left ventricular atrioventricular plane displacement: an echocardiographic technique for rapid assessment of prognosis in heart failure. AB - OBJECTIVE: To assess the prognostic value of atrioventricular plane displacement in heart failure patients. DESIGN: Patients were followed prospectively for one year after atrioventricular plane displacement determination. SETTING: Malmo University Hospital, with a primary catchment area of 250,000 inhabitants. PATIENTS: 181 patients with a clinical diagnosis of heart failure; age 75.7 (SD 5.2) years, duration of heart failure 2.7 (5.7) years; 100 men, 81 women. MAIN OUTCOME MEASURES: Mortality in relation to atrioventricular plane displacement. RESULTS: Total mortality was 22.7% (41/181), and was highly significantly (P = 0.001) related to atrioventricular plane displacement. Mortality within prospectively defined categories of displacement was: > or = 10.0 mm, 0% (0/19); 8.2 to 9.9 mm, 10.3% (3/29); 6.4 to 8.1 mm, 19.4% (12/62); and < 6.4 mm, 36.6% (26/71). The groups were similar in age, sex, angiotensin converting enzyme inhibitor and beta blocker treatment, and cause and duration of heart failure. CONCLUSIONS: Mortality in heart failure is strongly related to atrioventricular plane displacement. PMID- 9391284 TI - Prognostic implications of qualitative assessment of left ventricular function compared to simple routine quantitative echocardiography. AB - OBJECTIVE: To compare the prognostic value of qualitative estimates of left ventricular function with that of routine simple quantitative indices used in echocardiography. DESIGN: Retrospective follow up study. SETTING: University hospital. PATIENTS: The records of 2,964 patients who had undergone echocardiography and who could be traced on the family health services register were examined; 919 cases were included in the study, and a further 458 were used to validate the statistical models for prognostic assessment. There were 928 exclusions on the basis of referral for or diagnosis of alternative conditions, and 659 because of incomplete collection of the qualitative and quantitative data used in the study. MAIN OUTCOME MEASURE: Survival over the study period. RESULTS: A qualitative "eyeball" estimate of left ventricular function was of prognostic significance (relative risk of poor v good, 2.248; P << 0.001; 95% confidence interval 1.620 to 3.119). None of the quantitative echocardiographic indices was of independent prognostic significance when all variables were tested simultaneously in the regression model. CONCLUSIONS: A qualitative echocardiographic estimate of left ventricular dysfunction is of prognostic value, supporting the view of many cardiologists who use their overall impression of left ventricular function at echocardiography as the basis for treatment decisions. PMID- 9391285 TI - Long-term follow up of patients with implantable cardioverter-defibrillators and mild, moderate, or severe impairment of left ventricular function. AB - OBJECTIVE: To determine whether patients with life threatening ventricular tachyarrhythmias, impaired left ventricular function, and severe heart failure will benefit from implantable cardioverter-defibrillator (ICD) treatment. DESIGN: 410 patients were followed up after ICD implant. Left ventricular function was assessed by the New York Heart Association (NYHA) functional class of heart failure: 50 patients (12%) were in NYHA I-II, 151 (37%) in NYHA II, 117 (29%) in NYHA II-III, and 92 (22%) in NYHA III. Epicardial ICD implantation was performed in 209 patients (51%) and 201 patients (49%) received non-thoracotomy ICDs. RESULTS: Perioperatively, 12 patients (3%) died, more often after epicardial ICD implant (11/209 patients, 5%) than after transvenous implant (1/201 patients, < 1%) (P < 0.05). During a mean (SD) follow up of 28 (24) months (range < 1 to 114 months), 90 patients (23%) died: nine (2%) died from sudden arrhythmia; five (1%) also died suddenly but probably not from arrhythmic causes; 55 (14%) died from cardiac causes (congestive heart failure, myocardial reinfarction); 21 (5%) died from non-cardiac causes. The three year, five year, and seven year survival was 92-96% for arrhythmic mortality in NYHA class I, II and III, compared to a three year survival of 94% and a five year and seven year survival of 84% for patients in NYHA class II-III. 338 patients (82%) received ICD shocks (21 (SD 43) shocks per patient); patients in NYHA class II (83%), class II-III (84%), and class III (90%) received ICD discharges more often than those in class I-II (64%) (P < 0.05). The mean (SD) time interval between ICD implant and the first ICD shock was shorter in NYHA class II (16 (17) months), class II-III (19 (27) months), and class III (16 (19) months) than in class 0-I (22 (24) months) (P < 0.05). CONCLUSIONS: Patients with mild, moderate, and severe left ventricular dysfunction benefit from ICD treatment and these patients survive for a considerable time after the first shock. Survival is influenced by the degree of left ventricular dysfunction; aggressive treatment of heart failure is necessary as well as ICD therapy. PMID- 9391286 TI - Left atrial appendage function in patients with atrial flutter. AB - OBJECTIVE: To determine whether echocardiographic markers thromboembolic risk differ between patients with pure atrial flutter and patients with atrial flutter and intermittent atrial fibrillation. DESIGN: Patients with atrial flutter were followed up prospectively for 12 months to identify intermittent atrial fibrillation. After the follow up period, transthoracic and multiplane transoesophageal echocardiography were performed to assess left atrial chamber and appendage size, peak emptying velocities, and emptying fraction of the left atrial appendage. The presence of spontaneous echo contrast was also determined. SETTING: Tertiary cardiac care centre. PATIENTS: 20 consecutive patients with atrial flutter; 11 healthy subjects in sinus rhythm served as controls. RESULTS: Intermittent atrial fibrillation was documented in 11 patients by Holter monitoring or surface ECG; atrial fibrillation was not found in the other nine patients. Compared with the patients with pure atrial flutter, patients with atrial flutter and intermittent atrial fibrillation had larger left atrial chamber (mean (SD) 4.5 (0.6) v 3.8 (0.5) cm; 95% confidence interval 0.2 to 1.2; P = 0.01) and appendage areas (6.7 (2.2) v 4.8 (4.9) cm; 95% CI 0.4 to 3.5; P = 0.02), lower left atrial appendage emptying fractions (33 (11)% v 52 (11)%; 95% CI 8 to 29; P = 0.008), and also lower left atrial appendage emptying velocities (0.44 (0.21) v 0.79 (0.27) m/s; 95% CI 0.13 to 0.56; P = 0.005). In addition, a higher incidence of spontaneous echo contrast (11% v 36%) was observed in patients with atrial flutter and intermittent atrial fibrillation. CONCLUSIONS: Left atrial appendage function is depressed and spontaneous echo contrast more frequent in patients with atrial flutter and intermittent atrial fibrillation, as opposed to patients with pure atrial flutter. These data support the concept that patients with atrial flutter and intermittent atrial fibrillation have an increased risk of thromboembolic events and should therefore receive adequate anticoagulant treatment. PMID- 9391287 TI - Radiofrequency catheter ablation for idiopathic right ventricular tachycardia with special reference to morphological variation and long-term outcome. AB - OBJECTIVE: To assess the long term outcome of radiofrequency (RF) catheter ablation for idiopathic ventricular tachycardia (VT) originating from the outflow tract of the right ventricle, with special reference to the morphological variation in the VT-QRS complexes. PATIENTS: 13 patients whose ventricular tachycardia was treated with RF ablation were followed up more than 18 months after RF ablation. RESULTS: Endocardial mapping revealed the various extensions of ventricular tachycardia origin (from 0.5 x 0.5 cm to 2.0 x 2.0 cm) in which the earliest local electrogram was recorded during ventricular tachycardia. In all five tachycardias from a relatively wider origin (more than 0.5 x 0.5 cm) and in four of eight from a narrow origin (< 0.5 x 0.5 cm), subtle morphological variation in the VT-QRS complexes was observed. In tachycardias with morphological variation, the local electrogram at the tachycardia origin also showed concomitant variation in morphology and activation sequence. Ventricular tachycardia from a narrow site was eliminated by RF ablation to the confined site, but a larger number of RF applications was required in tachycardias from a wider origin. All 13 tachycardias were successfully ablated by RF current, and during the follow up period of 28.2 (SD 7.2) months, recurrence was observed in only one patient who had a wider origin. CONCLUSIONS: Long term efficacy of RF ablation was excellent in idiopathic ventricular tachycardia originating from the outflow tract of the right ventricle. Subtle morphological variations were frequently observed in this type of ventricular tachycardia, and about half of them represented a relatively wider arrhythmogenic area. PMID- 9391288 TI - Left atrial spontaneous echo contrast in patients with permanent pacemakers. AB - OBJECTIVE: To determine the relations between left atrial appendage function, spontaneous echo contrast, and thromboembolism in patients with different modes of permanent pacemakers. PATIENTS AND METHODS: 88 patients with pacemaker implantation and 25 healthy controls in sinus rhythm had transoesophageal echocardiographic examination of the left atrial appendage. Left atrial size, appendage area, peak filling and emptying velocities of the atrial appendage, and the presence or absence of spontaneous echo contrast and thromboembolism were determined. The results in 63 patients with ventricular pacing (group 1, subdivided into subgroup 1A: 42 patients with sinus rhythm, and subgroup 1B: 21 patients with atrial fibrillation) were compared with those in 25 patients with synchronous pacing (group 2), and 25 normal control subjects (group 3). RESULTS: Patients with ventricular pacing had two distinct appendage flow patterns: well defined biphasic filling and emptying waves in subgroup 1A, and irregular very low filling and emptying waves in subgroup 1B. The ejection fraction of the left atrial appendage in subgroup 1A was significantly better than that in subgroup 1B (mean (SD) 40.6 (12.0)% v 7.6 (5.0)%, P < 0.0001). The spontaneous echo contrast was observed in 90% of subgroup 1B patients but in only 19% in subgroup 1A (P < 0.05) and was not found in groups 2 and 3 (P < 0.0001). There was a trend for increased prevalence of spontaneous echo contrast in subgroup 1A v group 2 (P = 0.053). Thrombi were detected in two cases, and cardiogenic embolism occurred in one case in subgroup 1B. All patients with spontaneous echo contrast had ventricular pacing. Multivariate analysis showed that atrial fibrillation was associated with occurrence of spontaneous echo contrast in patients with ventricular pacing (P = 0.005). CONCLUSIONS: The left atrial appendage ejection fraction was lower with ventricular pacing than with synchronous pacing. With ventricular pacing there was a trend towards increased prevalence of left atrial spontaneous echo contrast in patients in sinus rhythm, and a significantly increased prevalence in patients with atrial fibrillation. PMID- 9391289 TI - QT dispersion as a risk factor for sudden cardiac death and fatal myocardial infarction in a coronary risk population. AB - OBJECTIVE: To test in a prospective study the hypothesis that increased QT dispersion in resting 12-lead ECG is a predictor of sudden cardiac death. DESIGN: A nested case-control study during a mean (SD) follow up time of 6.5 (2.8) years. SETTING: A prospective, placebo controlled, coronary prevention trial with gemfibrozil among dyslipidaemic middle aged men in primary (occupational) health care units: the Helsinki heart study. PATIENTS: 24 victims of fatal myocardial infarction, 48 victims of sudden cardiac death without acute myocardial infarction, and their matched controls. MAIN OUTCOME MEASURES: QT dispersion in baseline and pre-event electrocardiograms. RESULTS: At study baseline, QT dispersion was similar in all victims and controls. When estimated from the pre event ECG on average 14 months before death, the risk of sudden cardiac death in the highest QTPEAK (up to the peak of the T wave) dispersion tertile (> or = 50 ms) was 6.2-fold (95% confidence interval 1.7 to 23.5) compared with the risk in the lowest tertile (< or = 30 ms), and 4.9-fold (1.2 to 19.5) after adjustment for the presence of left ventricular hypertrophy, while QTPEAK dispersion could not predict fatal myocardial infarction. QTEND dispersion (up to the end of the T wave) in pre-event ECGs could not discriminate victims of either sudden cardiac death or fatal myocardial infarction from their matched controls. CONCLUSIONS: In middle aged men with a normal conventional QT interval in 12-lead resting ECG, increased QTPEAK dispersion is an independent risk factor for sudden cardiac death, but not for fatal myocardial infarction. PMID- 9391290 TI - Relation of serum cytokine concentrations to cardiovascular risk factors and coronary heart disease. AB - OBJECTIVE: To determine whether serum concentrations of the cytokines tumour necrosis factor alpha (TNF alpha) and interleukin 6 (IL-6), which regulate C reactive protein, are associated with cardiovascular risk factors and prevalent coronary heart disease. DESIGN: A population based cross sectional study. SUBJECTS AND METHODS: 198 men aged 50 to 69 years were part of a random population sample drawn from south London. Serum cytokine and C reactive protein concentrations were determined by enzyme linked immunosorbent assay. The presence of coronary heart disease was determined by Rose angina questionnaire and Minnesota coded electrocardiogram. RESULTS: Serum TNF alpha concentrations were positively related to body mass index and Helicobacter pylori infection, but inversely related to alcohol consumption. IL-6 concentrations were positively associated with smoking, symptoms of chronic bronchitis, age, and father having a manual occupation. TNF alpha was associated with increased IL-6 and triglycerides, and reduced high density lipoprotein cholesterol. IL-6 was associated with raised fibrinogen, sialic acid, and triglycerides. ECG abnormalities were independently associated with increases in IL-6 and TNF alpha, each by approximately 50% (P < 0.05 for TNF alpha, P < 0.1 for IL-6). The corresponding increases in men with an abnormal ECG or symptomatic coronary heart disease were 28% for TNF alpha and 36% for IL-6 (P = 0.14 for TNF alpha and P < 0.05 for IL-6). CONCLUSIONS: This study confirms that many of the phenomena with which C reactive protein is associated, are also associated with serum levels of cytokine, which may be the mechanism. PMID- 9391292 TI - Comparison of captopril and ibopamine in mild to moderate heart failure. AB - OBJECTIVE: To determine the effects of ibopamine 100 mg three times daily compared with captopril 25 mg three times daily on exercise capacity in patients with chronic heart failure. DESIGN: A randomised, double blind, parallel group comparison of the addition of ibopamine versus captopril during a period of 24 weeks. SETTING: 26 outpatient cardiology clinics in seven European countries. PATIENTS: 266 patients, with mild to moderate chronic heart failure (New York Heart Association (NYHA) functional class II, 81% and III, 19%) and evidence of an enlarged left ventricle. Patients received concomitant treatment with diuretics and/or digitalis. MAIN OUTCOME MEASURE: Exercise duration after 24 weeks of treatment, compared with baseline. RESULTS: Mean (SD) ejection fraction was 29 (8)% and the baseline exercise duration in the captopril and ibopamine groups 665 (160) and 675 (174) seconds, respectively. At the end of the study, exercise duration had improved in both groups, by 29 seconds in the ibopamine group (P < 0.01), and by 24 seconds in the captopril group (P < 0.05). There was no difference between groups (P = 0.69, 95% confidence interval -22 to 33). NYHA class, signs and symptoms score, and dyspnoea and fatigue index improved equally in both groups. The total number of adverse events was the same in both treatment groups, but gastrointestinal complaints occurred more often in the ibopamine group. The number of patients with premature withdrawals was no different. CONCLUSIONS: No difference was detected between the effect of captopril and ibopamine on exercise time in patients with mild to moderate heart failure during a treatment period of 24 weeks. PMID- 9391291 TI - Time dependent alterations of serum matrix metalloproteinase-1 and metalloproteinase-1 tissue inhibitor after successful reperfusion of acute myocardial infarction. AB - OBJECTIVE: To test the hypothesis that changes in serum matrix metalloproteinase 1 (MMP-1) and tissue inhibitors of metalloproteinase-1 (TIMP-1) after acute myocardial infarction reflect extracellular matrix remodelling and the infarct healing process. PATIENTS: 13 consecutive patients with their first acute myocardial infarction who underwent successful reperfusion. METHODS: Blood was sampled on the day of admission, and on days 2, 3, 4, 5, 7, 14, and 28. Serum MMP 1 and TIMP-1 were measured by one step sandwich enzyme immunoassay. Left ventricular volume indices were determined by left ventriculography performed four weeks after the infarct. RESULTS: Serum concentrations of both MMP-1 and TIMP-1 changed over time. The average serum MMP-1 was more than 1 SD below the mean control values during the initial four days, increased thereafter, reaching a peak concentration around day 14, and then returned to the middle control range. Negative correlations with left ventricular end systolic volume index and positive correlations with left ventricular ejection fraction were obtained for serum MMP-1 on day 5, when it began to rise, and for the magnitude of rise in MMP 1 on day 5 compared to admission. Serum TIMP-1 at admission was more than 1 SD below the mean control value, and increased gradually thereafter, reaching a peak on around day 14. Negative correlations with left ventricular end systolic volume index and positive correlations with left ventricular ejection fraction were obtained for serum TIMP-1 on days 5 and 7, and for the magnitude of rise in TIMP 1 on days 5 and 7 compared to admission. CONCLUSIONS: Both MMP-1 and TIMP-1 showed significant time dependent alteration after acute myocardial infarction. Thus MMP-1 and TIMP-1 may provide useful information in evaluating the healing process as it affects left ventricular remodelling after acute myocardial infarction. PMID- 9391293 TI - Early changes in left ventricular anterior wall dynamics and coordination after coronary artery surgery. AB - OBJECTIVE: To study how asynchronous left ventricular wall motion changes early after uncomplicated coronary artery surgery. DESIGN: A prospective study done before, and at 0.5, 1, and 3 hours after coronary artery grafting, with intraoperative transoesophageal cross sectional guided M mode echocardiograms, high fidelity left ventricular pressure, and thermodilution cardiac output measurements. The extent and velocity of left ventricular anterior wall thickening were measured, along with regional work and power production. Abnormal thickness changes during the isovolumic periods were detected, and their effect on energy transfer quantified as cycle efficiency. SETTING: Tertiary referral cardiac centre. PATIENTS: 25 patients with a history of chronic stable angina, mean (SD) age 60 (9) years with three vessel coronary artery disease, undergoing uncomplicated coronary artery bypass grafting. RESULTS: 4 patients had primary incoordination, as shown by wall thinning during isovolumic contraction and delayed onset of thickening (group A), and nine had premature thickening due to incoordination elsewhere (group B). The extent (thickening fraction 43 (12)% v 73 (19)%) and velocity (1.7 (0.4) v 2.5 (0.6) cm/s) of thickening were reduced in group A v group B (P < 0.001), as were regional stroke work (2.2 (0.8) v 3.3 (0.4) mJ/cm2) and peak power production (19 (5) v 32 (7) mW/cm2), P < 0.05. In group A, these values all increased significantly within 30 minutes of operation. In group B, the extent of wall thickening and peak power production were unaffected by surgery, though cycle efficiency and regional stroke work both improved by 30 minutes v before operation (73 (9)% v 61 (8)%, 4.5 (0.9) v 3.3 (0.4) mJ/cm2, P < 0.01). Surgery had no consistent effect on left ventricular cavity size, shortening fraction, or cardiac output in either group. CONCLUSIONS: Even in the absence of evidence of overt ischaemia, major disturbances of ventricular synchrony--both regional and generalised--are present in patients with a history of chronic stable angina requiring coronary artery bypass grafting. They regress within 30 minutes of revascularisation, suggesting that they are the direct result of coronary stenosis. PMID- 9391294 TI - Anatomical criteria for the diagnosis of sinus venosus defects. AB - BACKGROUND: The diagnosis of sinus venosus defects remains a matter of debate. It is crucial to provide solid anatomical criteria, by identifying the very nature of the atrial septum relative to sinus venosus defects, to diagnose and differentiate them from other interatrial communications. OBJECTIVE: This study was designed to reestablish the anatomical criteria for the diagnosis of sinus venosus defects. METHODS: Five specimens with sinus venosus defects from the cardiopathological museum were examined. Study of the abnormal hearts was supplemented by examining the extent and structure of the atrial septum in 10 normal hearts. The echocardiograms and surgical notes were reviewed from 18 patients seen between July 1991 and August 1996 at the Royal Brompton Hospital in London diagnosed preoperatively to have a sinus venosus defect. RESULTS: The nature of the oval fossa and its muscular borders were identified in the normal hearts. In all three autopsied specimens of the superior variety of sinus venosus defect, the mouth of the superior caval vein was overriding the intact muscular anterosuperior border of the oval fossa. Two specimens thought initially to have the inferior variety of sinus venosus defect were re-classified as having defects within the oval fossa as it was the deficient oval fossa itself, rather than its intact muscular border, that was overridden by the mouth of the inferior caval vein. Sixteen patients had been diagnosed echocardiographically as exhibiting the superior variant of the defect. Retrospective review showed overriding of the superior caval vein across the upper rim of the oval fossa in 12 patients. These findings were confirmed by surgery in 11 patients with the 12th awaiting operation. Overriding of the fossa by the caval vein was not found in the other four patients. Surgery in all of these showed the defect to be within the oval fossa. In two patients diagnosed echocardiographically as having inferior defects, the surgical findings confirmed a biatrial connection of the inferior caval vein in one patient, the findings in the second were equivocal. CONCLUSIONS: The key anatomical criterion for the diagnosis of sinus venosus defects is overriding of the mouth of the superior or inferior caval vein across the intact muscular border of the oval fossa. The interatrial communication is then formed within the mouth of the overriding vein, and is outside the confines of the oval fossa. PMID- 9391295 TI - Quantitative, non-invasive assessment of ventricular septal defect shunt flow by measuring proximal isovelocity surface area on colour Doppler mapping. AB - OBJECTIVE: To determine whether the proximal isovelocity surface area (PISA) method could be applied to estimate the magnitude of ventricular septal defect (VSD) shunt flow. DESIGN: Prospective analysis of clinical, echocardiographic, and angiographic data. SETTING: University hospital. PATIENTS: 14 children with VSD. METHODS: Colour Doppler images of VSD shunt flow were obtained in parasternal long axis view, four chamber view or both, adjusted to provide the best imaging of flow. The VSD shunt flow rate and shunt volume were calculated as follows: shunt flow rate (SFR) = 2 pi r2 V/BSA in ml/s/m2; shunt volume = SFR x shunt duration time. The shunt volume, shunt fraction, and pulmonary to systemic flow ratio (Qp:Qs) were confirmed by cardiac catheterisation. RESULTS: There was a correlation between shunt variables determined by PISA and those by catheterisation, including shunt volume (r = 0.78, P = 0.001) and shunt fraction (r = 0.74, P = 0.003). Qp:Qs was also significantly correlated with SFR (r = 0.79, P = 0.0007). The SFR was significantly different between the four patients with Qp:Qs < 2.0 (mean (SD) 54 (33) ml/s/m2) and the 10 patients with Qp:Qs > 2.0 (186 (69) ml/s/m2) (P = 0.004). CONCLUSIONS: These data suggest that the PISA method is a reliable non-invasive investigation for the quantitative assessment of VSD shunt flow and provides important information for decisions regarding surgical repair. PMID- 9391296 TI - Two teenagers, traumatic aortic disruption, and transoesophageal echocardiography. AB - A report of two teenagers is presented who were passengers of the same vehicle when it was involved in a serious road traffic accident. Their vehicle was hit on the side at high speed by a second vehicle. Both teenagers sustained multiple injuries. On arrival at a district general hospital they required resuscitation, exploratory laparotomies, and postoperative intensive care. Neither had radiographic evidence to suggest any mediastinal injury, but transoesophageal echocardiography demonstrated aortic disruption in both patients. They were transferred to a regional cardiothoracic centre where their lesions were successfully repaired and both have made a full recovery. PMID- 9391297 TI - Concrete induced cardiac contusion. AB - A previously fit 22 year old man was struck in the chest by a concrete block dropped through the windscreen of his car while he was driving on the motorway. He suffered extensive chest wall trauma and lung contusion, which subsequently precipitated acute respiratory distress. On admission ECG showed right bundle branch block and left axis deviation. Three days later QRS duration was normal but there was anterior ST segment elevation and subsequent T wave change. There was a large rise in creatine kinase, and echocardiography revealed septal and apical hyokinesis as well as a mobile mass attached to the left side of the interventricular septum, which had the echogenic texture of myocardium. The patient had fixed perfusion defects in the areas of hypokinesis on thallium scanning but the coronary arteries were unobstructed at angiography. He was treated with warfarin in the short term and an angiotensin converting enzyme inhibitor in the longer term and has made an asymptomatic recovery. Outpatient echocardiography two months after the injury demonstrated some recovery in overall left ventricular systolic function and no evidence of the intracardiac mass. This case illustrates some of the typical features of non-fatal cardiac contusion associated with non-penetrating cardiac trauma, and was complicated by partial thickness avulsion of a strip of the myocardium in the interventricular septum. PMID- 9391298 TI - Traumatic ventricular septal defect. AB - A 26 year old man was admitted to hospital following a traffic accident. He had been sitting in the back of a car without wearing a seat belt. He suffered crush injuries on the anterior chest wall, trunk, and legs. On admission he was awake and cooperative, but restless, and obviously in severe pain. Radiography of the skull, facial bones, chest, spine, pelvis, and legs revealed a shaft fracture of the left femur and tibia and fracture of the 7th and 8th right ribs. The patient was transferred to the University Hospital of Zurich for further assessment and surgical repair of the lower limb fractures three days later. Because of worsening clinical condition with onset of partial respiratory insufficiency and new loud systolic murmur at the left sternal edge, a transthoracic echocardiography was performed, which showed an apical ventricular septal defect. Surgery was performed immediately. The ventricular septal defect was successfully repaired using a Teflon felt patch and interrupted sutures with pledgets, and sealed with glue. At six months' follow up the patient was doing well. Ventricular septal defects after blunt chest trauma occur either because of heart compression between sternum and the spine or because of myocardial infarction. In the present case the ventricular septal defect appeared three days after the accident, probably secondary to a post-traumatic myocardial infarction. Patients with blunt chest trauma and suspicion of cardiac contusion should be monitored carefully. PMID- 9391300 TI - Flecainide levels--a cautionary note. PMID- 9391299 TI - Iatrogenic atrioventricular bypass tract following a Fontan operation for tricuspid atresia. PMID- 9391301 TI - Prognostic significance of ST-T segment alterations in patients with non-Q wave myocardial infarction. PMID- 9391302 TI - Stent placement in the outlet of the right ventricle. PMID- 9391303 TI - Raised plasma BNP in a patient with acute pulmonary thromboembolism. PMID- 9391304 TI - Assessment of endothelial function using plasma markers. PMID- 9391305 TI - [Newer approach of screening test for antinuclear antibodies: an enzyme-linked immunosorbent assay detecting antinuclear antibodies characteristic of connective tissue diseases]. AB - An enzyme-linked immunosorbent assay (ELISA) has been developed for the detection of antinuclear antibodies (ANAs) previously established as diagnostic and/or prognostic marker ANAs for various connective tissue diseases. The antigen used in ELISA is a mixture of purified recombinant or natural antigens including single-and double-stranded DNA, RNP, Sm, SS-A/Ro, SS-B/La, centromere, topoisomerase I and Jo-1 antigens. Thirty hundred and fifty nine patients sera from a variety of connective tissue diseases and 113 normal human sera (NHS) were examined. ELISA ANAs were positive in 3.5% of NHS and 80.2% of patients sera at cut off index 11.5, whereas indirect immunofluorescent antinuclear antibodies (FANAs) using HEp-2 cells were positive in 9.7% of NHS and 92.5% of patients sera at 1:160 serum dilution. More than 80% of sera from systemic lupus erythematosus, mixed connective tissue disease and primary Sjogrens disease were ELISA ANAs positive. Mean value of ELISA ANAs was highest in sera of patients with MCTD. ELISA ANAs were positive in 92.5% of sera with marker ANAs for connective tissue diseases. Mean value of ELISA ANAs was higher in sera with more than two marker ANAs than in sera with a single ANA or in sera without marker ANAs. In contrast incidence and mean value of ELISA ANAs were low in sera positive for anti topoisomerase I antibody or anti Jo-1 antibody. Sensitivity, specificity and agreement (accuracy) for connective tissue diseases with marker ANAs were as follows: ELISA ANAs (at index 11.5): 92.5%, 88.3% and 90.9%: FANAs (at 1:160 serum dilution): 99.0%, 70.4% and 88.1%, respectively. ELISA ANAs, thus, are specific for connective tissue diseases when compared to FANAs and previous ELISA for the detection of total ANAs. Moreover, ELISA ANAs are able to measure precise ANAs titers and are much less labor intensive when screening a large number of clinical specimens. PMID- 9391306 TI - [2-5 AS activity in serum and peripheral blood mononuclear cells for chronic active hepatitis C and the relationship to clinical outcome of interferon therapy]. AB - Of 49 patients with chronic hepatitis C treated by interferon (IFN), we measured 2'-5'-oligoadenylate synthetase (2-5 AS) activity in peripheral blood mononuclear cells (PBMC) and serum before and after the IFN therapy and studied the correlation with the clinical outcome. Before IFN therapy, the levels of 2-5 AS in PBMC and serum were significantly higher in patients with chronic hepatitis C than in healthy controls, though there was no correlation between the 2-5 AS activity and the clinical outcome. When PBMC were stimulated with IFN in vitro, the induced 2-5 AS activities in patients with chronic hepatitis C were almost same as those in healthy controls. Among patients infected with hepatitis C virus (HCV) genotype II which was considered relatively resistant to IFN, patients whose HCV was disappeared from serum by IFN therapy showed good induction of 2-5 AS activity by IFN in vitro, whereas patients in which serum HCV remained positive after the therapy showed poor response to IFN in vitro. The levels of 2 5 AS in PBMC 2 months after IFN therapy were still higher in patients whose HCV was continuously disappeared from serum by the therapy (complete remission) than in healthy controls. The in vitro induction of 2-5 AS in patients whose HCV in serum remained positive after the therapy was significantly lower than in patients with complete remission. The induction of 2-5 AS activity in patients to whom IFN therapy was ineffective, significantly decreased after the IFN therapy as compared with the activity measured before the therapy. These findings suggest that measurement of 2-5 AS activity in PBMC in vitro through IFN therapy might be useful for predicting in vivo responsibility to IFN and also knowing the change of the responsibility. PMID- 9391307 TI - [A case of congenital complete heart block in a mother with anti-52 kD SS-A/Ro antibodies]. AB - We report a case of congenital complete heart block (CCHB). A 38-year-old woman was admitted our hospital because of fetal bradycardia at 21 weeks 3 days of gestational age. She had no symptom of collagen disease. On admission, laboratory data showed positive anti-nuclear antibodies, anti-SS-A/Ro antibodies and anti-52 kD SS-A/Ro antibodies. But anti-60 kD SS-A/Ro and anti-SS-B/La antibodies were negative. Consequently anti-52 kD SS-A/Ro antibodies positive woman had an infant with CCHB. The baby was equipped with pacemaker at the age of 2 months. This report suggests that anti-52 kD SS-A/Ro antibodies may play an important role in the development of CCHB. PMID- 9391308 TI - [Pernicious anemia associated with chronic thyroiditis and suspected latent adrenal insufficiency]. AB - A 64-year-old female referred to our hospital because of severe anemia. Peripheral blood examination showed macrocytic anemia; red blood cell count was 1.49 x 10(6)/microliters, hemoglobin concentration was 5.6 g/dl, hematocrit was 16.1% and MCV was 108 fl. Serum VB 12 level was significantly low as 58 pg/ml. Upper gastrointestinal examination disclosed chronic atrophic gastritis. Anti intrinsic factor and anti-parietal cell antibodies were detected in the serum and Schilling's test was positive. Thus a diagnosis of pernicious anemia was made. Though the serum free T 3 and free T 4 levels were in normal ranges, the elevated serum TSH and positive tests for anti-microsome and anti-thyroglobulin antibodies indicated that the patient had chronic thyroiditis. Then other endocrinological examinations were performed. Low level of urinary 17-OHCS and a hypo-reactive pattern of rapid ACTH test led to a diagnosis of latent adrenal insufficiency. This case could be categorized into polyglandular autoimmune syndrome. PMID- 9391309 TI - [A case of hemophagocytic syndrome with severe liver injury manifestating adult Still's disease]. AB - In April 1988, a 23 year-old woman developed high fever, arthralgia, eruptions and splenomegaly. She was treated with non Steroid anti-inflammatory drugs, and the symptoms disappeared. In June 1991, she was diagnosed as adult Still's disease and treated with prednisolone. In July 1994, she was treated with pulse therapy methylprednisolone due to high fever, eruptions, arthralgia and the high levels of ferritin. However, due to the marked increase of serum transaminase and bilirubin levels, she was referred to University hospital. She developed hepatic failure after admission Bone-marrow puncture revealed hemophagocytosis. She died ten days after admission. She was diagnosed as hemophagocytic syndrome combined with acute hepatic failure. PMID- 9391310 TI - [A case of reactive arthritis fallen after shigellosis infection]. AB - A 25-year-old woman was admitted for general arthralgia in July, 1989. Reactive arthritis with arthralgia after Shigellosis was diagnosed by sex, localization of arthralgia and positive for HLA-B 27. Within 3 weeks after starting diclifenac sodium 75 mg/day, the arthralgia remitted. It has been reported that patients who are positive for HLA-B 27 have a more severe acute or chronic sacroiliitis, and our case may support this report. PMID- 9391311 TI - [A case of Wegener's granulomatosis associated with refractory bowel granulomatous ulcers]. AB - We describe a 58-year-old woman who developed Wegener's granulomatosis (WG) complicated by a perforation of the transverse colon caused by necrotizing granulomatous vasculitis. In addition, her colon lesion continued in spite of high dose corticosteroid and cyclophosphamide therapy. She was admitted to our hospital because of her severe tonsillitis in Dec., 1994. She was diagnosed as having WG because she had oral ulcer, antibiotics-resistant lung infiltration, renal dysfunction and positive C-ANCA. Just after we started high dose steroid therapy, the transverse colon was perforated because of vasculitis, and she underwent emergency operation. Many vasculitic lesions were found in the small intestine, colon, and mesenterium. The disease was improved by corticosteroid and cyclophosphamide therapy except for a sustained ulcer with necrotizing vasculitis in the sigmoid colon region even 1 year after the operation. Although WG rarely complicates digestive tract lesions as initial manifestations, they reach 12% of the causes of death of WG in Japan. Therefore, we should take care of digestive tract lesions when we follow-up patients with WG. PMID- 9391312 TI - [Survival of Vibrio cholerae non-O1 in freshwater river]. AB - The survival of Vibrio cholerae non-O1 was investigated in sterile and untreated river water. The essential nutrients for growth of the organism were also investigated using a compound medium. V. cholerae non-O1 was shown to increase in autoclaved sterile river water, but did not increase in filtrated river water, and starting with an initial number of 10(6) CFU/ml organism, the organism could not be isolated from untreated river water after day 10. The growth or survival of the organism was also studied with filtrated river water to which all essential nutrients except one was added. In this water, V. cholerae non-O1 did not increase when either phosphate or a carbon source was not present. These results indicate that V. cholerae non-O1 can survive in river water by taking nutrients for growth from solutes and particle matter in the river water. PMID- 9391313 TI - [A survey of cognitive probability structure of risk of death by cause]. AB - A person is believed to choose a health related behavioral alternative based on his/her perception of the health risk. This survey tried to clarify the structure of perceptions relating to risk of death, which seems to be the most fundamental among various health risks. A survey was performed in 1995 on 350 employees of two major companies. Subjects were shown two paired causes of death and asked which diseases caused deaths more frequently in the Japanese. Diseases included cancer, cerebral apoplexy, heart disease, traffic accident, tuberculosis, and lung cancer. The results obtained were as follows: 1. Generally, the order of perceived risk for each cause of death was similar to the real death numbers. 2. While traffic accidents tended to be overestimated for its risk, the risk perception for heart disease and cancer tended to be underestimated. 3. Young people tended to overestimate the risk of traffic accidents more than older people. Based on these results, it is suggested that people tend to overestimate current risks, and to underestimate future risks. This may contribute to the improvement of risk communication between health care providers and residents/patients. PMID- 9391314 TI - [Association of participation in health checkups with medical service utilization in a rural town]. AB - Association of participation in health checkups in 1983-94 with medical service utilization in 1989-94 was studied. The study subjects consisted of 1,198 males and 1,247 females aged 30 years or over, who were members of the National Health Insurance in a rural town in Kyoto prefecture. One hundred and four males and 95 females died during 1989-94. Cumulative death rates were higher among both males and females with less participation in checkups than among those with higher participation, however, there was no difference seen for medical service utilization. Regarding subjects alive in 1994, both males and females who participated frequently in checking in 1983-88 also visited medical facilities more frequently and had higher medical costs in 1989-94 than those with less frequent participation. Similar analysis for checkups in 1989-94, showed that only females had that tendency. Among participants in checkups in 1989-94, there were smaller numbers of hypertensives, current smokers, and current drinkers than among non-participants, therefore, the participants are thought to have decreased their risk for chronic diseases. In conclusion, frequent participation in health checkups are thought to increase medical utilization even with a lower prevalence or risk of chronic diseases. PMID- 9391315 TI - [Health conditions and life styles of residential elderly. Part 1. Characteristics and factors related to being healthy elderly persons from a survey of health life style]. AB - A survey was conducted in a highland community of Gifu prefecture on 815 residential elderly persons, over 65 years old, utilizing self-administered questionnaires to study the degree of association between life style factors and health conditions. A total of 718 valid participants (90.6% of the total) were analyzed and cross-examined by applying Breslow's seven health factors, daily behavior factors, physical health consciousness, social mobility factors (13 behavior factors which were determined by the Institute of the Elderly) to determine associations with respondents health conditions. Of the participants, 310 were male and 408 were female with mean ages of 73.8 +/- 6.9 years old and 74.0 +/- 6.9 respectively. The survey results indicated that there were no prominent gender differences over the mobility rate and physical health consciousness among participants. However, it was also found that in the male participants the per person prevalence of past history of illness was higher than in females. In the daily activity analysis, female participants indicated a higher prevalence of difficulty in walking and balance maintenance while ascending and descending stairs. In the general daily activity analysis, male participants had a higher requirement for an aide. In social mobility, male participants recorded a high degree of self-reliance by being physically, socially, and intellectually active and the female participants by fulfilling their social responsibilities. Breslow's seven health indicators were applied and their evaluation points were calculated as 3.9 +/- 1.7 for male participants and 3.5 +/- 1.8 for female participants which showed a significantly higher score for the male participants. The survey also indicated that factors which increase the Breslow score for males, were having a spouse, hobbies, a satisfactory life, a habit of exercising, and non-smoking which in females an appropriate sleeping habit (not less than 7 hours), and working, were the positively related factors. PMID- 9391316 TI - [Support plan for children with chronic intractable diseases and their family- results of inquiries about patients and their family]. PMID- 9391317 TI - [Analysis of Vi antigen genes of Salmonella typhi and study of its regulation]. PMID- 9391318 TI - [Studies on the regulation of toxin production in Clostridium perfringens]. PMID- 9391319 TI - [Environmental regulation of the fimbriae and toxin expression in Actinobacillus actinomycetemcomitans]. PMID- 9391320 TI - [Current topics of pathogenicity in Salmonella]. PMID- 9391321 TI - [Staphylococcal leukocidin and gamma-hemolysin]. PMID- 9391322 TI - [Characteristics of the cellulolytic ruminal bacterium Fibrobacter succinogenes and its attachment to cellulose]. PMID- 9391323 TI - [The GTP-binding proteins as targets for bacterial toxins]. PMID- 9391325 TI - [Clinical study of the application of the concepts of systemic inflammatory response syndrome (SIRS) in liver cirrhosis with or without hepatocellular carcinoma with upper gastrointestinal hemorrhage--a retrospective study]. AB - The retrospective study was aimed at assessing the usefulness of the application of the criteria of SIRS for identifying a subset of patients with higher mortality in 22 cases (21 patients) of liver cirrhosis with upper gastrointestinal hemorrhage (GIH) and 16 cases (14 patients) of liver cirrhosis with hepatocellular carcinoma with upper GIH. The following results were obtained; (1) The incidence of SIRS on upper GIH was 66%. (2) The mortality rate in patients with SIRS on GIH was significantly higher than in patients with non SIRS on GIH in 60 days after GIH was significantly higher than in patients with non-SIRS on GIH in 60 days after GIH (50% vs. 8%; p < 0.01). (3) The rate of patients who met four criteria of SIRS on GIH or during admission and of patients whose durations of SIRS was over 5 days was significantly higher in the died patients with SIRS on GIH than in the survived patients with SIRS on GIH (67% vs. 0%; p < 0.01, 67% vs. 0%; p < 0.01, respectively). These results suggested that the application of the criteria of SIRS was useful for identifying a subset of patients with higher mortality in chronic liver disease with GIH. PMID- 9391324 TI - [Studies on bacillary dysentery cases of overseas travellers--during 1979 to 1995]. AB - A total of 36,780,440 overseas travellers during 1979-1995 (17 years) were quarantined at Osaka and Kansai Airport Quarantine Station, 84,777 travellers reported themselves suffer from diarrhoea. Stools from 29,587 persons were bacteriologically examined. Various enteropathogenic bacteria were isolated from 9,766 (33.0%) patients of the stools examined. Isolated species were as follows: Plesiomonas shigelloides (3,234 cases); Salmonella spp. (2,236 cases); enterotoxgenic Escherichia coli (1,621 cases); Vibrio parahaemolyticus (1,959 cases); and Shigella spp. (1,242 cases). 1,278 different Shigella strains were isolated from 1,242 cases who were thus diagnosed as bacillary dysentery patients. The suspected regions or countries for infection of these cases were analysed. The serovars and antibiotic-sensitivities of the isolated strains were examined. Colicine typing of S. sonnei strains were also done. The results can be summarized as follows. 1) The most cases (53.4%) were infected in India. 2) The percentage distribution of sub-species of the strains was as follows; S. sonnei (57.8%), S flexneri (29.8%), S. boydii (8.4%), and S. dysenteriae (4.0%), respectively. 3) The major colicine type of S. sonnei strains were type 6 and 0. 4) The percentage of Antibiotic-resistant strains of each sub-species was S. dysenteriae (92.2%), S. sonnei (89.4%), S. flexneri (87.1%), and S. boydii (84.9%), respectively. The percentage of Antibiotic-resistant strains of S. flexneri were increased annually. PMID- 9391326 TI - [Pathological and immunohistochemical analysis of giant cells of pancreas]. AB - Multinucleated giant cells in the pancreas (five giant cell carcinomas, a mucinous cystadenocarcinoma attended with many osteoclast-like giant cells, 42 invasive ductal carcinomas and 29 chronic pancreatitises) were examined. Three types of multinucleated giant cell were identified: epithelial type, coexpressive type, mesenchymal type. Epithelial type expressed epithelial markers, such as keratin and EMA in 23 ductal carcinomas. Coexpressive type expressed both epithelial markers and mesenchymal marker vimentin was in four ductal carcinomas. Mesenchymal type expressed mesenchymal markers, vimentin and CD68 in four osteoclastoid type giant cell carcinomas, the mucinous cystadenocarcinoma, six ductal carcinomas and ten chronic pancreatitises. Epithelial and coexpressive type were considered to be epithelial neoplastic origin, those had bizarre appearance and transitional area from definite adenocarcinoma area. Vimentin expression is associated with sarcomatous proliferation. Mesenchymal type was considered to be nonneoplastic and a certain type of macrophage polykaryons. PMID- 9391327 TI - [A case of solitary gastric varices treated with B-RTO method followed by MR angiography]. PMID- 9391328 TI - [A case of colitic cancer arising from villous type of dysplasia]. PMID- 9391329 TI - [Sigmo-sigmoid fistula formation in a case of ulcerative colitis]. PMID- 9391330 TI - [A case of metastatic carcinoma of the duodenum from bladder]. PMID- 9391331 TI - [A case of drug-induced liver injury caused by Saiko-Keishi-Kankyoto with thrombocytopenia]. PMID- 9391332 TI - [An autopsied case of adult onset Still's disease accompanied by fatal liver failure, and simultaneously complicating adenocarcinomatous peritonitis from unidentified primary site]. PMID- 9391333 TI - [A case of autoimmune hepatitis associated with anti-phospholipid antibody syndrome]. PMID- 9391334 TI - [A case of double cholecysto-duodenal fistula associated with gallstone ileus]. PMID- 9391335 TI - [A case of anaplastic carcinoma of the pancreas, disclosed a hemosuccus pancreaticus]. PMID- 9391336 TI - [Successful treatment of pancreatic pseudocyst with octreotide; a case report]. PMID- 9391337 TI - [Evaluation of modified rapid urease test (MR urea S) in diagnosis of Helicobacter pylori (HP)]. PMID- 9391338 TI - [The discovery and molecular mechanism of protein splicing]. PMID- 9391339 TI - [Natural killer T cells: their origin and functions]. PMID- 9391340 TI - [Human NK inhibitory and activating receptors that bind to human leukocyte antigens (HLA) regulate NK cells and T cells]. PMID- 9391341 TI - [Development of animal cloning by nuclear transfer]. PMID- 9391342 TI - [Organic solvent tolerance in Escherichia coli]. PMID- 9391343 TI - [Calorimetry of proteins (II)]. PMID- 9391345 TI - 'The evidence suggests that ...'. PMID- 9391346 TI - Paving the way: stepping stones to evidence-based nursing. AB - Evidence-based practice is an emerging paradigm in health care. This paper outlines the main features of this paradigm and its potential value to nursing. Evidence-based practice is based on a conceptual framework that examines the extent of evidence available in support of particular clinical practices. The Quality of Evidence Ratings adapted by the National Health and Medical Research Council (NHMRC) from the United States Preventive Services Task Force are discussed, and the strengths and weaknesses of different categories of evidence are highlighted. Potential barriers to implementation of research into practice are identified. The authors suggest that legal, ethical, economic and humane imperatives oblige nursing to develop evidence-based practice as one of several viable contributions to nursing knowledge. Suggestions for analysing current research and for the planning of the direction of future nursing research are made. PMID- 9391347 TI - The acceptance of a philosophically based research culture? AB - This paper addresses a trend that emerged at a nursing research conference held in New South Wales during 1996. The trend was a downplaying of the relevance of philosophical and theoretical bases for research in nursing. Such a trend would not be of concern if all research aimed to merely generate local, non generalizable conclusions and recommendations. However, research that aims to develop knowledge that can provide relevant knowledge on a broader, or universal scale, needs to be based on coherent and relevant philosophical and/or theoretical foundations. To adequately critique such research it should ideally be linked to a philosophy or theory that situates it in both historical and intellectual loci and consequently, the raison d'etre for the research should arise. This paper begins with an exploration of Immanuel Kant's philosophy and proceeds to a discussion on the philosophical underpinnings of Dorothea Orem. It is intended that such discussion will assist the clinician to see the relevance that philosophy has to clinically based research in nursing and, importantly, the critique of such research. In conclusion, two possible reasons are suggested as to why philosophy and theory in nursing research may appear to be increasingly frowned upon. It is hoped that a greater understanding of the power of philosophically based research will contribute to an increased enthusiasm and uptake of philosophically based research projects by nurses. PMID- 9391348 TI - Nursing, a bridge to peace in our region: opinions on mutual co-operation between Israeli and Palestinian professional nurses and nurse educators. AB - The purpose of this study was to assess the activities and future direction of mutual co-operation of professional nurses from Israel and Judea and Samaria. Questionnaires were distributed to the participants, nurse educators and professional nurses from Judea and Samaria who were attending a workshop in clinical instruction at the Assaf HaRofeh School of Nursing, in Israel. Most of the nurses polled were interested in continued educational dialogue and expanded educational programmes in various nursing specialties both in education and service. The areas that received priority included: intensive care, operating room and emergency room nursing. Areas that received lower priorities included; midwifery, geriatrics and nephrology nursing. The participants expressed positive reactions to continued joint educational activities such as full length courses, workshops, day seminars and joint educational projects. The results showed an overwhelming interest for continued professional ties. Areas in the fields of clinical expertise and nursing education reflected the needs of their respective communities. PMID- 9391349 TI - The elderly adult in the emergency department. AB - Over 65-year olds were nominated by emergency staff at a Melbourne regional hospital as a patient group of particular concern. With nurse academics from Monash University, a collaborative study was undertaken of elderly patients and the circumstances of their attendance. The focus of the study was on those elderly patients who were triaged as non-acute and who may have been disadvantaged by the priority given to acute cases. The triage records obtained over a 5 month period were analyzed, and a survey administered to selected patients. Over 65-year olds were found to constitute 19% of incoming patients. They figured more prominently in urgent triage categories than those under 65 years of age, were more likely to be referred by a health professional, and more likely to be admitted or transferred. There was no evidence to suggest slower progress through the emergency department for the non-acute elderly than for their counterparts under 65 years of age. PMID- 9391350 TI - Breaking a social silence: registered nurses share their stories about tertiary nursing education. AB - This research presents some of the qualitative findings from a mixed methods study exploring the personal and professional effects of tertiary education for post registration women nurses. The researcher combined both qualitative and quantitative approaches. The specific methods employed were survey questionnaire, 'journalling' and interviews. The results demonstrated that the participants were able to identify and verbalise personal and professional experiences associated with tertiary study, through multiple voices. A distinctive theme of empowerment emerged in their stories. However, by sharing their stories, it was evident that encounters in their lives were enmeshed with dialectical relationships. In conclusion, the associated tension that characterized their personal and professional relationships, indicated that these women could not control their social worlds and that their experiences were integrally linked to their engendered nature. PMID- 9391351 TI - Role identity and job satisfaction of community health nurses. AB - Although many efforts have been made to articulate the nature of community health nursing practice, there continues to be a great deal of confusion about the roles and functions of community health nurses. This article reports a descriptive research study of the role identity and job satisfaction of 43 staff community health nurses practicing in home health care, generalized public health nursing, combined public health/home health care (PH/HHC), and specialized programmes. Although differences were found, the home health, generalized public health, and PH/HHC nurses shared a similar core identity. The major differences were seen for the nurses working in specialized programmes. No significant differences were found in job satisfaction among the four groups. PMID- 9391352 TI - The specialist learning disability nurse in Wales. AB - This paper reflects on the development of services for people with learning disabilities within the United Kingdom and focuses on the role of the specialist nurse. The nurse's contribution to the care of this client group has been the subject of debate and controversy for a number of years. Developments within the learning disability filed in Wales in particular are explored, with the All Wales Strategy for People with Learning Disabilities providing the background and context for these developments. An example of how specialist nurses from around Wales came together in order to share good and best practice is discussed. The conclusion is that the specialist learning disability nurse has a great deal to offer, but must be prepared to work together with other professionals as well as service users and their families. PMID- 9391353 TI - A therapeutic programme for people with dementia. AB - A programme involving Tai Chi and structured reminiscence was trialed with nine people suffering from moderately advanced dementia. The analysis reported here aimed to examine stories the people told with a view to understanding the purpose of story telling in their lives. Themes derived from the narrative data had a strong evaluative quality, ranging from simple evocative expressions to more cognitive complex insights or treasures. The study indicated a major aim in story telling as being able to generate life values, both for the enrichment of identification of self, and to pass on or leave for today's youth. Findings here further substantiate Luke and Freiden's view that old age has its own cognitive and spiritual goals to achieve. There is strong evidence that people with moderate dementia still aim to participate in that endeavour. PMID- 9391354 TI - Where does rehabilitation fit in the picture of care for the trauma patient? PMID- 9391355 TI - The history of the Glasgow Coma Scale: an interview with professor Bryan Jennett. Interview by Carole Rush. AB - An interview with one of the founders of the Glasgow Coma Scale provides a partial history of the development and dissemination of this assessment tool. Professor Bryan Jennett credits nurses with the rapid spread and universal acceptance of the scale. PMID- 9391356 TI - Attitudes and beliefs of Afro-Americans related to organ and tissue donation. AB - A descriptive study was conducted to identify problems associated with the attitudes, beliefs, and lack of participation of a small Afro-American population concerning organ donation. I. King's theory of goal attainment was used to compare stated beliefs with reported behaviors that would indicate a willingness to participate in organ donation. PMID- 9391358 TI - Termination of resuscitation for traumatic cardiac arrest. PMID- 9391359 TI - Easing pain in children. AB - The assessment and management of pain in children has been essentially ignored until recently. Thankfully, these "dark ages of pain" are ending. The trauma nurse is an integral part of the pain management team and can have a positive impact on outcome by using a combination of relatively simple strategies. These include using multiple types of assessment to measure the severity of pain; providing adequate pain relief with a combination of pharmacologic and nonpharmacologic interventions; and carefully monitoring and documenting the efficacy of all pain management approaches. PMID- 9391360 TI - Helpful Websites. PMID- 9391361 TI - Babygard infant transport. PMID- 9391362 TI - Relationship of breastfeeding and formula-feeding practices with infant health outcomes in an urban poor population. AB - The article reports a study examining symptoms of infection and use of medications and the health care system by breastfeeding or formula-feeding urban poor mothers. A prospective, self-report design was used. Mothers completed a demographic and anthropometric questionnaire, an infection checklist, and a medication and health care system survey. Results showed that more of the breastfeeders were white, older, and economically better off than formula feeders. Scores on the infection checklist were higher for those feeding their infants by bottle. Colds, rashes, episodes of vomiting, ear infections, colic, and health care utilization were less frequent for breastfed infants. This small study suggests that there is a protective effect of breastfeeding in this population and provides a basis for larger epidemiologic and cross-sectional studies. PMID- 9391363 TI - Breastfeeding education for early discharge: a three-tiered approach. AB - Discharge teaching has presented many challenges since 48-hour stays have become routine for uncomplicated obstetric patients and their term infants. Sleepy infants, lack of maternal psychologic readiness, and lack of a full milk supply may frustrate efforts to teach breastfeeding during this limited time frame. The key is to develop a three-tiered teaching approach that includes affective, psychomotor, and cognitive learning during the antepartum, immediate postpartum, and long-term postpartum course. This approach focuses on: (1) the mother's critical concerns and the infant's critical biologic needs during breastfeeding initiation and (2) a collaborative approach that enhances the overall success of breastfeeding continuation. PMID- 9391364 TI - Combining breastfeeding and employment: increasing success. AB - As more families recognize the tremendous advantages of breastfeeding, more mothers are choosing to continue breastfeeding when they return to an employment setting. Using a chronological format, this section describes strategies health care providers can use to assist employed mothers to prepare for continued breastfeeding, develop skills needed to continue breastfeeding, overcome challenges, and successfully meet their breastfeeding goals. Numerous charts/tables that can be shared with mothers are included. PMID- 9391365 TI - Comparison of confidence between mothers who breastfed and formula fed their preterm infants. AB - Mothers develop confidence to care for their preterm infants in challenging situations. To date no studies have explored the effects of choice of feeding method on the development of maternal confidence in the preterm population. A descriptive, longitudinal design was used to explore the demographic characteristics of a convenience sample of 173 maternal-preterm infant dyads to examine whether confidence increased over time and to compare levels of confidence between mothers who formula fed and mothers who breastfed their preterm infants. There were no significant differences between feeding groups in maternal confidence; however, maternal confidence within each group increased significantly over time. PMID- 9391366 TI - Cup feeding the newborn: what you should know. AB - Cup feeding is gaining increased recognition as an alternative method of feeding infants breast milk. For the term infant, cup feeding is suggested when the mother is unavailable to put the infant to breast. In the preterm population, cup feeding may be initiated before the preterm infant is ready to be put to the breast. Although further research is needed to substantiate the proposed benefits of cup feeding, the article provides information about selecting the appropriate candidates for cup feeding and offers step-by-step instructions to ensure that cup feeding is done safely. PMID- 9391367 TI - Maternal feelings after cessation of breastfeeding: influence of factors related to employment and duration. AB - The purpose of this study was to measure feelings following weaning in women planning employment within 1 year postpartum and to examine the effects of factors related to duration and employment on these feelings. No significant differences in feelings of sadness, anger, guilt, and relief were reported by women who weaned due to mother-led reasons or baby-led reasons or in women who did or did not meet their breastfeeding goal. Women who did not feed their babies as planned when employed, however, felt significantly more sadness/depression and guilt compared to women who achieved their planned method of feeding. PMID- 9391368 TI - Minimizing infant exposure to and risks from medications while breastfeeding. AB - The advantages of breastfeeding to the mother and newborn are many. Lactating mothers frequently ask about the safety of taking medications and the risk to their newborn. It is well established that all drugs are excreted into breast milk. However, most medications appear in only small amounts within the breast milk. With the availability of numerous resources on drug use while breastfeeding, a medication can be identified as contraindicated or compatible with breastfeeding. By understanding the anatomy of the breast, principles of lactation, and drug passage into breast milk, an approach to minimize the transfer of the medications in the breast milk to the newborn can be developed. The plan should usually support and encourage the mother to continue to breastfeed her infant. PMID- 9391369 TI - Nurse practitioner descriptions for primary care centers: opportunities for ownership. AB - The drastic shift in health care delivery and the accompanying emphasis on health care outcomes has contributed to a rapid proliferation of nurse practitioner services. Prepared as advanced practice nurses with specific assessment skills, primary care nurse practitioners have the opportunity to be participant players rather than observers in business negotiations. To gain a market share of the expanding field, these nurses must assert their position on establishing primary care centers. This qualitative study focused on nurse practitioner descriptions of their needs in a university primary care group practice. Four major response patterns, Vision, Structure, Incentives, and Significance, were clarified by multiple themes defined by explanatory elements for a research-based topology to guide nurse practitioners and schools into positions of ownership of primary care centers in interdisciplinary partnerships. PMID- 9391370 TI - Put prevention into practice. Alcohol and other drug abuse. PMID- 9391371 TI - A comparison of conventional percussion and auscultation percussion in the detection of pleural effusions of hospitalized patients. PMID- 9391372 TI - Dermatology on the Internet. PMID- 9391373 TI - Clinical health problem: failure to thrive. AB - Families in which NOFTT is present need interventions that target behaviors and identify stressors that contribute to decreased caloric intake. A holistic approach to the situation is required, to deal with considering the problem which may stem from multiple sources. Maternal perceptions of health and diet can be influenced by the practitioner in a sensitive manner, encouraging balance. Infant feeding difficulties can be identified by the practitioner and appropriate referrals can be made to FTT clinics that are experienced in working with these infants and their caregivers. Public health nurses can be utilized to further assess families, follow up on health teaching, and provide referrals to community resources to alleviate stressors. Management of NOFTT by a practitioner in the primary care setting is feasible and cost-effective. PMID- 9391374 TI - After four years, it's back to being 'just' a member. Interview by Tina Steger Gratz. PMID- 9391375 TI - AIM--helping Michigan children get their immunizations. PMID- 9391376 TI - Legislative liaison: is it for me? PMID- 9391377 TI - Survey results offer insight. PMID- 9391378 TI - Direct Medicare reimbursement for NPs, CNSs takes effect Jan. 1. PMID- 9391380 TI - Nursing reaches out to the media. PMID- 9391381 TI - Cost or quality? PMID- 9391379 TI - ANA quality indicators: meaningful measurement. PMID- 9391382 TI - For health-care reform, call a nurse. PMID- 9391383 TI - Trends affecting nursing and patient care in hospitals. PMID- 9391384 TI - Acute care surgical nurses during downsizing: stress, self-esteem, and social intimacy. PMID- 9391385 TI - ASTP addresses transplant issues for decade ahead with position paper . PMID- 9391386 TI - Chloramines in municipal water: considerations for dialysis facilities. PMID- 9391387 TI - Getting it right: how to design your water treatment system from the ground up. PMID- 9391388 TI - Drug resistant organisms and their implications for the outpatient dialysis setting. AB - Ever since antibiotics have been used to combat infection, resistance to them has also developed. The elderly and immunosuppressed populations found in today's dialysis facility comprise a group at risk for acquiring infection caused by drug resistant organisms. By utilizing antibiotics prudently and developing infection control strategies based on the disease/organism epidemiology, mode of transmission and individualized needs of the patient as well as the care setting, the proliferation of drug resistant organisms may be curtailed. PMID- 9391389 TI - Study points to latex glove permeability. PMID- 9391390 TI - Anemia guidelines suggest iron as missing link in managing hematocrits . PMID- 9391391 TI - National program hopes to improve outcomes for ESRD patients with indicator tool. PMID- 9391392 TI - Hiring renal RNs: start with love, compassion. PMID- 9391393 TI - Local networks essential for success in implementing global capitation. PMID- 9391394 TI - Looking to the future for renal care. Randolph highlights plans for one-year term as NRAA president. PMID- 9391395 TI - Rehabilitation on the other side of the world. Hong Kong encourages ESRD patients to stay healthy with exercise. PMID- 9391396 TI - Fitness training: an ongoing effort. PMID- 9391397 TI - Telemedicine project looks to improve ESRD care, lower costs. TRC/Georgetown University Medical Center begin two-year experiment. PMID- 9391399 TI - ASN at 30. PMID- 9391398 TI - Focus on pre-dialysis logical, but how will nephrologists define it? PMID- 9391400 TI - Council of American Kidney Societies predicts "critical shortage" of nephrologists by 2010. PMID- 9391401 TI - Why choose nephrology? PMID- 9391402 TI - Is there a nephrologist in the house? PMID- 9391403 TI - How will we manage ESRD patients in the future? PMID- 9391404 TI - Rekindling the flame. PMID- 9391405 TI - Control of drug resistant organisms in the outpatient dialysis unit. PMID- 9391406 TI - Reshuffling the ESRD food chain. PMID- 9391407 TI - Bureaucracy a major discouragement in getting ESRD patients back to work. PMID- 9391408 TI - IDPN: is there a future? PMID- 9391409 TI - What are the options beyond IDPN? PMID- 9391410 TI - Latest look at Canadian register data shows lower mortality among PD patients. PMID- 9391412 TI - Dearing. So what? PMID- 9391413 TI - Lesson-planning. PMID- 9391411 TI - Canadian Association to launch on-line dialysis course in January '98. PMID- 9391414 TI - Marshalling the arguments for community practice teachers. PMID- 9391415 TI - Choosing your first post. PMID- 9391416 TI - Health promotion in professional practice. PMID- 9391417 TI - Recording what you learn from reading. PMID- 9391418 TI - What's the point of ... systematic reviews? PMID- 9391419 TI - Gems of thought from Florence Nightingale. PMID- 9391420 TI - Gelastic epilepsy. A case study of neurological disorder. PMID- 9391421 TI - Discipline and interpersonal relations in operation theatre. PMID- 9391422 TI - SCNA peer assistance program in nursing. PMID- 9391423 TI - Promoting ethics in the acute care setting. PMID- 9391424 TI - Trust, too valuable to lose. PMID- 9391425 TI - Tips on getting a good start in a new job. PMID- 9391426 TI - Enhancing student learning and employee health through university-community partnerships. PMID- 9391428 TI - Spiritual care: a challenge for the occupational health nurse. PMID- 9391427 TI - Ambulatory care: changing roles for school nurse. PMID- 9391429 TI - Depositions: what you need to know. PMID- 9391430 TI - Writing for publication: a primer. PMID- 9391431 TI - Medicare reimbursement for NPs and CNSs to be a reality, January 1, 1998. PMID- 9391432 TI - Nursing ethics at the juncture of two kinds of care. PMID- 9391433 TI - Theories of justice and care: a paradigm for nursing ethics. PMID- 9391434 TI - New frontiers in health care decision-making: information, decision-making, and divided loyalties. PMID- 9391435 TI - Failure of the cure model in intensive care. PMID- 9391436 TI - Calling all schools of nursing: we want to know about death. PMID- 9391437 TI - Legal nurse consulting in South Carolina--emergence of a nursing specialty and an innovative educational program. PMID- 9391438 TI - Making the decision to change careers. PMID- 9391440 TI - Patient advocates speak out about unsafe care and whistle-blowing retribution. PMID- 9391439 TI - The return of health care inflation meets preventive care. PMID- 9391441 TI - The Wellness celebration: a community-wide approach to health promotion. PMID- 9391442 TI - Greenwood nurses support a partnership of caring people. PMID- 9391443 TI - Don't become extinct, wear a helmet! PMID- 9391444 TI - Caring for the Korean-American community. PMID- 9391445 TI - Parish nursing: caring for the community. PMID- 9391446 TI - SCNA papin column: hope for addicted nurses. PMID- 9391447 TI - The seduction of Ethel. PMID- 9391448 TI - Not for profit or for profit health care does it really matter? PMID- 9391449 TI - Human relations weakness--a leading cause of unsuccessful job campaigns. PMID- 9391450 TI - Survey results SCNA Ethics Committee. PMID- 9391451 TI - Nurses express views on health care reform. PMID- 9391453 TI - SCNA CEAC Column: selling nursing. PMID- 9391452 TI - Transfusion of red blood cells/platelets. The State Board of Nursing for South Carolina. PMID- 9391454 TI - PAPIN column--program for impaired nurses. PMID- 9391455 TI - The Cowboy Model of reform. PMID- 9391456 TI - Janice Weinberg's Career Clinic: mail vs. telephone--how should you initiate contact with your targeted employers? PMID- 9391457 TI - The impact of managed care on a state agency. PMID- 9391458 TI - The times they are a changin'. PMID- 9391459 TI - The role of the clinical nurse specialist in a managed care environment. PMID- 9391460 TI - Patient Safety Act. PMID- 9391461 TI - The times they are a'changing. PMID- 9391462 TI - The chemically dependent nurse and intervention. PMID- 9391463 TI - Managed care or terrorism? You be the judge. PMID- 9391464 TI - Are you committing strategic resume errors that can sabotage your job-search campaign? PMID- 9391465 TI - Advanced practice nursing. PMID- 9391466 TI - Work force diversity: nursing perspectives. AB - All too frequently, diversity is viewed negatively or, at best, neutrally. The challenge of diversity is one of embracing it. The extent to which we are successful in meeting this challenge will determine South Carolina's and America's competitiveness and effectiveness in health care and in the global marketplace. Although much of the focus of this article has been work force diversity within nursing, attention to diversity is applicable within all health services markets, where human resources is the primary conduit for progress. Therefore, elements of this report can be generalized to all health care providers, where racial under-representation, in comparison to the populations served, is present. PMID- 9391468 TI - Nursing education in transition. PMID- 9391467 TI - "Promoting cultural competence in the baccalaureate nursing student". PMID- 9391469 TI - Nursing workforce planning: the RWJF Colleagues in Caring Project. PMID- 9391470 TI - RNs voice critical concerns about patient care. PMID- 9391471 TI - What is N-STAT? PMID- 9391473 TI - How to get elected to a state position voted on by the entire SC General Assembly. PMID- 9391472 TI - The Patient Safety Act of 1996 (H.R. 3355). PMID- 9391474 TI - Office of public health nursing--SC DHEC. PMID- 9391475 TI - Clinical ethics and the South Carolina nurse. PMID- 9391477 TI - Emerging infectious diseases: a challenge to nurses. PMID- 9391476 TI - Medical reform: the long and short of it. PMID- 9391478 TI - Disease reporting: the first step in surveillance. PMID- 9391479 TI - HIV/AIDS in aging: concerns and challenges. PMID- 9391480 TI - Scientific and cultural exchange trip to China. PMID- 9391481 TI - Who's taking care of mama? PMID- 9391482 TI - Delegation: concepts and decision-making process. PMID- 9391483 TI - American Nurses Association position statement on home care for mother, infant and family following birth. PMID- 9391484 TI - HIV, hepatitis--B, hepatitis--C, blood-borne diseases nurses' risks, rights, and responsibilities. PMID- 9391485 TI - Clinical ethics and the South Carolina nurse. PMID- 9391486 TI - Re-engineering--threat of opportunity? PMID- 9391487 TI - Educational technologies in the information age. PMID- 9391488 TI - Nursing informatics--application to clinical practice. PMID- 9391489 TI - Information management in nursing. AB - The information technology environment is here to stay. Computers are the devices being used to manage the extension of care out of hospitals and into the community. Nurses are the focal point of clinical documentation because they access patient data most often. Nurses know what information is needed and seek ways to collect that information. Nurses can dictate the type of data collected and how that data is presented to allow the most appropriate, well-informed decisions possible. Nurses have the knowledge to make decisions, and with information management skills, we will have the means to communicate that knowledge to others. PMID- 9391490 TI - A practice model for nursing in developmental disabilities. PMID- 9391491 TI - Applications for ultrasonography in the emergency department. AB - The primary emergency applications of diagnostic ultrasonography are now reasonably well defined. Basic characteristics of the emergency department examination, including its rapid and highly focused nature, determine both the limitations and advantages of this technique. As greater experience and more advanced technologies develop in the emergency department setting, the role of ultrasonography will likely expand into areas that are not generally practiced today. Doppler technology, especially, promises to provide the emergency department physician of the future an even more powerful tool for the rapid diagnosis of a variety of common and critical conditions. PMID- 9391492 TI - Cardiac ultrasonography. AB - The use of cardiac ultrasonography in the emergency department currently has two primary indications: determining the presence of cardiac tamponade and determining cardiac activity in patients with apparent pulselessness. Physicians are able to recognize both conditions after appropriate, but not overextensive, training sessions. The evaluation of penetrating trauma, acute myocardial infarction, and severe shock of uncertain origin require a higher degree of training and experience. All of these conditions are discussed in this article. PMID- 9391493 TI - Ultrasonography in blunt abdominal trauma. AB - This article discusses studies of the use of ultrasound in patients with blunt abdominal trauma, both in initial assessment and ongoing evaluation. Reviews of studies of children and adults to detect the presence and extent of hemoperitoneum and organ injuries are presented. Ultrasound results are compared with diagnostic peritoneal lavage, computed tomography, clinical course, and autopsy results. The central question addressed is to what extent can ultrasonography replace or supplement other techniques, particularly diagnostic peritoneal lavage, in the assessment of patients with blunt abdominal trauma. Ultrasound equipment, technique, scoring scales, limitations, and training issues are also addressed. PMID- 9391494 TI - Pelvic ultrasonography. AB - Pelvic ultrasonography is a valuable tool for the emergency physician in the evaluation of the wide spectrum of pelvic complaints presenting to the emergency department. The goal of this article is to outline pelvic problems that can be readily identified by the emergency physician using pelvic sonography early in the patient's evaluation. A special emphasis is placed on the sonographic diagnosis of ectopic pregnancy. PMID- 9391495 TI - Abdominal ultrasonography. AB - This article focuses on ultrasonographic examinations of the abdomen and important intra-abdominal pathology. The liver and biliary tree are discussed first, followed by the use of ultrasonography in diagnosing appendicitis, ascites, and bowel obstruction. Pyloric stenosis and intussusception, important pediatric intra-abdominal problems, are also discussed. PMID- 9391496 TI - Vascular ultrasonography. AB - Although vascular ultrasonography has been established as an essential diagnostic tool in many clinical settings, its role in the emergency department patient population is uncertain. Preliminary reports of emergency physician--directed ultrasonography are promising. Further studies are needed to establish its reliability and suitability in the emergency department setting. PMID- 9391497 TI - Renal ultrasonography. AB - Renal US is one of several imaging modalities available to the emergency physician in the evaluation of patients with acute urologic disorders. It offers excellent anatomic detail without exposure to radiation or contrast agents but does not assess renal function. It is particularly useful in the evaluation of renal colic, although its role here may decrease with increasing availability of helical CT. It also has an important role in excluding bilateral renal obstruction as the cause of acute renal failure. Doppler renal US is likely to take on a more prominent role in the evaluation of renal trauma and is the diagnostic study of choice to rule out renal vein thrombosis. Bedside emergency renal US performed and interpreted by emergency physicians with limited training and experience is gaining in use and acceptance. Its role at present is primarily to identify unilateral hydronephrosis in patients with suspected renal colic. This role is likely to expand in the future as emergency US use grows and technology advances. Bedside emergency renal US may eventually be used in the evaluation of patients with acute renal failure, suspected renal vein thrombosis, and renal trauma. PMID- 9391498 TI - Ultrasound detection of foreign bodies and procedure guidance. AB - As emergency physicians become familiar with the use of ultrasonography, this safe procedure will likely become a standard technique having multiple uses in the emergency department. Ultrasonography assists in foreign body localization and retrieval and is potentially important in applications, such as reliable endotracheal tube placement, visualization of ingested medication, vascular access, and drainage of collected fluids. PMID- 9391499 TI - Credentialing issues in emergency ultrasonography. AB - The use of ultrasonography, traditionally performed by radiologists, is becoming increasingly widespread in emergency medicine. Consequently, much debate has evolved over whether emergency medicine physicians are qualified to provide this service, and the criteria by which training and credentialing can be achieved. This article discusses training and credentialing guidelines, paths to becoming credentialed in emergency sonography, and quality assurance issues. Also, strategies are proposed for emergency departments seeking to perform emergency sonography. PMID- 9391500 TI - Mibefradil--a new calcium-channel blocker. PMID- 9391501 TI - Two new retinoids for psoriasis. PMID- 9391502 TI - Emerging and resurgent infections. PMID- 9391503 TI - Measuring hygiene practices: a comparison of questionnaires with direct observations in rural Zaire. AB - To date questionnaire surveys have been the most commonly used instruments to measure hygiene behaviours related to water and sanitation. More recently, a number of studies have used structured observations to study practices related to diarrhoea. During a trial of a hygiene education intervention to reduce diarrhoea among young children in Bandundu, Zaire, both instruments were used to measure the disposal of child faeces and various hand-washing practices. Three hundred families were observed and follow-up interviews performed with 274 (91%) mothers. At the individual level, agreement between observed and reported behaviour was little better than might be expected by chance. There was evidence of over reporting of hand-washing before food preparation (44% vs 33%; P = 0.03), hand washing before eating (76% vs 60%; P < 0.001) and disposal of the child's faeces in a latrine (75% vs 40%; P < 0.001). On the other hand, hand-washing before feeding the child was reported less often than it was observed (7% vs 64%; P < 0.001). Our data are consistent with the hypothesis that, in general, mothers over-report 'desirable' behaviours. At the same time, our data indicate that open questions may lead to under-reporting of certain behaviours. The repeatability of observations at both the individual and population levels remains to be established. PMID- 9391504 TI - Issues in the design and interpretation of studies to evaluate the impact of community-based interventions. AB - Increasingly, epidemiologists are faced with the need to evaluate the impact of an intervention that is delivered at the level of a community or cluster of individuals, rather than at the individual level. This has profound implications for the design and interpretation of a study to evaluate its impact. We start by discussing the issues arising in the extension of the randomized double-blind controlled trial methodology to the evaluation of interventions delivered to clusters of individuals, or to whole communities, where the unit of randomization is a cluster of individuals rather than an individual. We then consider alternative approaches to design, discuss their relative strengths and weaknesses and present a framework of design options. Finally we propose a pragmatic approach to evaluation design in this setting. We believe that the answer lies in the judicious selection of different design elements, combined in such a way that when the evidence from each is presented together, a clear picture of the impact of the intervention emerges. We illustrate this using an example from the recent literature. PMID- 9391505 TI - Incidence estimates of late stages of trachoma among women in a hyperendemic area of central Tanzania. AB - The purpose of this study is to estimate 5-year incidences of conjunctival scarring and trichiasis, and 10-year incidence of corneal opacities due to trachoma, using prevalence data from a population sample of 6038 women living in a trachoma-hyperendemic area of central Tanzania. Previous surveys have documented the age-specific prevalence of scarring, trichiasis, and corneal opacities in women in hyperendemic areas. Using the age-stratified prevalences of these different clinical signs, corresponding incidence rates were estimated. Transition rates from one sign to the next were also obtained by restricting the risk group to only women with a specific trachoma sign. Thus, the 5-year incidence of trichiasis among women with conjunctival scarring, and the 10-year incidence of corneal opacities among women with trichiasis were estimated. Incidences of all the signs markedly increased with age. For scarring, 5-year incidence rates increased from 3.1% in the 15-19 age category to 14.3% for women between 55 and 59 years. The 5-year incidence of trichiasis ranged from 0.3% in the 15-19 age category to 7.5% in the age group 55-59. Corneal opacities due to trachoma were highest in the age group 45-54; the 10-year incidence increased to 2.8%. The 5-year incidence of trichiasis among only women with scars increased from 3.2% in the 15-19 age group to 15.1% in women in the 55-59 age group. Once trichiasis is present, almost one-third of the women below 35 and more than 40% of the women older than 45 will develop corneal opacities in a 10-year interval. These estimates are important in understanding the dynamics of progression of trachoma from conjunctival scarring to the potentially blinding signs of trichiasis and corneal opacities. They provide important information for planning adequate services in areas where trachoma is endemic and surgery for trichiasis is a key factor to avoid blindness from trachoma. They also provide clues to the pathogenesis that may be useful in the development of new methods of control. PMID- 9391506 TI - Nationwide prevalence study of hypertension and related non-communicable diseases in The Gambia. AB - The prevalence of hypertension, diabetes and obesity in The Gambia was assessed in a 1% population sample of 6048 adults over 15 years of age, 572 (9.5%) subjects were hypertensive according to WHO criteria (a diastolic blood pressure (DBP) of 95 mmHg or above and/or a systolic blood pressure (SBP) of 160 mmHg or above); 325 (5.4%) had a DBP of 95 mmHg or above, and 39 (2.3%) a DBP of 105 mmHg or above; 428 (7.1%) had a SBP of 160 mmHg or above. By less conservative criteria (a DBP of 90 mmHg or above and/or SBP of 140 mmHg or above), 24.2% of subjects were hypertensive. The prevalence of hypertension was similar in the major ethnic groups and in urban and rural communities. Age and obesity were risk factors for hypertension; female sex was an additional risk factor for diastolic hypertension. Several communities had a prevalence of diastolic hypertension double the national rate, and significant community clustering of diastolic hypertension (P < 0.01) was confirmed by Monte Carlo methods. Genetic and/or localized environmental factors (such as diet or Schistosoma haematobium infection), may be involved 140 (2.3%) subjects were obese. Obesity was associated with female sex, increasing age, urban environment, non-manual work and diastolic hypertension. Only 14 (0.3%) subjects were found to be diabetic. Hypertension appears to be very prevalent in The Gambia, with a substantial population at risk of developing target organ damage. Further studies to delineate this risk and appropriate interventions to reduce it are needed. PMID- 9391507 TI - Resistance of falciparum malaria to chloroquine and sulfadoxine-pyrimethamine in Afghan refugee settlements in western Pakistan: surveys by the general health services using a simplified in vivo test. AB - Surveys of drug resistant falciparum malaria were conducted in several Afghan refugee settlements, distributed over a 700 km range in western Pakistan, during the transmission seasons of 1994 and 1995. Symptomatic malaria patients were recruited by a process of passive case detection at the refugees' basic health units. To facilitate follow-up by local health workers, a modified version of the WHO extended in vivo test was adopted in which blood smears were taken from each subject, and clinical symptoms recorded, at weekly intervals. Resistance to chloroquine and sulfadoxine-pyrimethamine was identified in every settlement. The frequency of chloroquine resistance ranged from 18% to 62%. Resistance occurred mostly as RI, with RII resistance never exceeding 11%. Resistance to sulfadoxine pyrimethamine occurred at much lower frequencies, ranging from 4% to 25%. There was a resumption of clinical symptoms at the onset of parasite recrudescence in over 90% of cases. The policy of using chloroquine as first-line treatment might be changed in favour of sulfadoxine-pyrimethamine in most camps and areas of western Pakistan. The modified in vivo test was almost as accurate as the normal WHO in vivo test in identifying the grade of resistance, and should prove a useful tool for the monitoring of resistance to common antimalarials by district health services. PMID- 9391508 TI - Optimizing the malaria data recording system through a study of case detection and treatment in Sri Lanka. AB - The potential of using malaria incidence data routinely collected from endemic regions for disease control and research has increased with the availability of advanced computer-based technologies, but will depend on the quality of the data itself. We report here an investigation into the relevance of malaria statistics provided by the routine data collection system in Moneragala, a rural malaria endemic region in Sri Lanka. All patients (n = 321) treated for malaria in 2 clusters of health care centres (HCCs) of both the private and the public sector in the administrative regions of Moneragala and Buttala Divisional Secretariat (D.S.). Divisions were studied in December 1995/ January 1996. The catchment area of these HCCs included a population resident in 53 Grama Niladhari (GN) areas, the smallest administrative units of the country. Almost equal numbers of malaria patients were detected and treated at Government and private health care institutions, and in 70% of them treatment was based on a diagnosis confirmed by microscopy. The routine data recording system, however, included only statistics from the Government sector, and only of patients whose diagnosis was microscopically confirmed. In compiling data, the origin of a case of malaria is attributed to the D.S. Division in which the institution (at which the patient was treated) was located, rather than the area in which the patient was resident, which was inaccurate because 90% of malaria patients sought health care at institutions located closest to their residence, thus crossing administrative boundaries. It also led to a loss of resolution of spatial data because patients' addresses recorded at the Government HCCs to the village-level are replaced in the statistics by the D.S. Division, which is a coarse spatial unit. Modifications to the system for malaria case recording needed to correct these anomalies are defined here. If implemented, these could result in major improvements to the quality of data, a valuable resource for the future of malaria control. The paper reiterates the call for the use of a standard spatial unit within a country to facilitate exchange of data among health and other sectors for the control of tropical diseases. PMID- 9391509 TI - Konzo associated with war in Mozambique. AB - We report an epidemic of konzo, symmetric spastic paraparesis associated with cassava consumption and cyanide exposure: 384 patients were treated in rehabilitation centres; the prevalence rate in a badly affected area was 30/1000. Most patients were children over 3 and women. Owing to war, communities turned to bitter cassava as their staple and took shortcuts in its processing. When the war ended, they continued to depend on inadequately processed bitter cassava. The epidemic lasted 2 years (the last year of war and the first of peace) with peaks each year during the cassava harvest. Although most cases were reported from rural inland areas, patients also came from small towns and the coast. School children had raised urinary thiocyanate and linamarin and low inorganic sulphate concentrations. Urinary thiocyanate values were lower than those previously reported in konzo epidemics, probably because we collected specimens before the cassava harvest and epidemic peak. The necessary conditions for konzo were present: intensive cultivation of bitter casava, insufficient processing, a probable high cyanide intake, and a low intake of protein-rich foods. PMID- 9391510 TI - High prevalence of mutations in the dihydrofolate reductase gene of Plasmodium falciparum in isolates from Tanzania without evidence of an association to clinical sulfadoxine/pyrimethamine resistance. AB - Recently the efficacy of sulfadoxine/pyrimethamine (S/P) in treatment of uncomplicated falciparum malaria in Tanzania has been seriously compromised by the development of resistance. The occurrence of active site mutations in the Plasmodium falciparum gene sequence coding for dihydrofolate reductase (DHFR) is known to confer resistance to pyrimethamine. This study investigates the occurrence of these mutations in infected blood samples taken from Tanzanian children before treatment with S/P and their relationship to parasite breakthrough by day 7. The results confirm the occurrence of one or more DHFR mutations in all the samples, but no relationship was found with the presence of parasites in the blood at day 7. The results suggest that alterations in the coding region for dihydropteroate synthetase (DHPS), the enzyme target for sulfadoxine, should be studied in order to predict resistance to the S/P combination. It has been proposed earlier that sulfadoxine could itself act on DHFR, because of a false dihydrofolate produced by drug metabolism through DHPS and dihydrofolate synthase. The results of this treatment study suggest that such a possibility is unlikely. PMID- 9391511 TI - Maternal nutrition and socio-economic status as determinants of birthweight in chronically malnourished African women. AB - The birthweight is the most important determinant of mortality and morbidity in the neonatal period and may have an influence on health in adult life. The high rate of low birthweight in developing countries is therefore a major health problem. Maternal malnutrition is usually assumed to be a causal factor but other environmental factors are also involved. In this study we analysed maternal nutritional and socio-economic factors as determinants of birthweight in term infants from a rural African society characterised by a high rate of chronic malnutrition. Relations of maternal weight, gestational weight gain, parity, socio-economic status and infant sex with birthweight were analysed in 1,477 women and child pairs. The selected women were followed from early pregnancy and had an uncomplicated delivery at term of a living singleton child. The gestational weight gain was 5.6 (SD 6.0) kg and the mean birthweight 2.933 kg (SD 408). Maternal weight, representing the maternal long-term nutritional situation, was the most important independent determinant of birthweight, accounting for 13.0% of the variance in birthweight. The weight gain, representing the short term nutritional situation, explained only 5.6% of the variance. Birthweight increased by 20 g (CI 18-23) for each kg maternal weight and by 15 g (CI 12-18) for each kg gestational weight gained. The socio-economic difference in birth weight was 153 g (CI 109-196) 88 of which (CI 48-128) remained unexplained after adjustment for differences in maternal weight, parity and gender. Improved long term nutritional situation and living conditions seems to be the most important prerequisites to counteract low birthweight in developing countries. PMID- 9391512 TI - Typhoid fever in Ujung Pandang, Indonesia--high-risk groups and high-risk behaviours. AB - We performed a hospital-based case-control study to identify high risk groups and routes of transmission of typhoid fever in the city of Ujung Pandang on the island of Sulawesi, Indonesia. The annual incidence of this disease in southern Sulawesi is estimated at 3.1/1000 and the case fatality at 5.1% Cases were 50 patients over 13 years of age admitted to Stella Maris Hospital with a diagnosis of typhoid fever between June and September 1991. Diagnosis was made on clinical grounds and in 90% of cases confirmed by a Widal test. Controls were 42 patients admitted for non-infectious disorders during the same period and individually matched by age and sex. Controls did not have a history of typhoid fever. Interviews took place in hospital. Analysis was by unconditional logistic regression. High-risk groups consisted of those who were single, unemployed and those who had a university education. Median age of cases was 22 years. Consumption of food from warungs (food stalls in the street) was strongly associated with risk (OR = 45). Both cases and controls washed hands after use of the toilet and before meals, but cases used soap significantly less often (OR = 30). The results of this study can be used to take preventive measures against this severe disease of educated and single young adults by targetting them for IEC-activities emphasizing the importance of thorough hand-washing and the need to take care in the selection of street-foods. PMID- 9391513 TI - Evaluation of Amplicor- and IS6110-PCR for direct detection of Mycobacterium tuberculosis complex in Singapore. AB - One hundred and seventy-eight samples from 168 individuals were tested for Mycobacterum tuberculosis complex (Mtc) using Amplicor PCR, IS6110-PCR (in house), acid fast (AF)-staining and culture. Thirty-one samples were positive by culture, but 37 samples were later resolved to be truly positive for Mtc. Of these, Amplicor detected 32 (86.5%), IS6110-PCR detected 31 (83.6%), and AF staining 21 (56.8%). None of the 141 Mtc-negative samples was positive by these tests, thus giving 100% specificity. Although the IS6110-PCR was more sensitive than Amplicor in detecting spiked Mtc DNA, it was not more sensitive than the latter in detecting Mtc in clinical samples. Reasons likely to account for the PCR false negativity were (i) sample inoculum size, (ii) nonuniform samples due to clumping effect of Mtc and (iii) the absence of target gene sequences for IS6110-PCR. Culture negativity, on the other hand, was likely to be associated with nonviable Mtc. Amplicor PCR is promising for direct detection of Mtc. The IS6110-PCR, however, may not be as suitable because of possible existence of IS6110-deleted Mtc strain in Singapore. PMID- 9391514 TI - Conference report: international conference on HIV and iron, Brugge 1997. PMID- 9391515 TI - A time to eradicate and a time to control. PMID- 9391516 TI - Human behaviour and cultural context in disease control. PMID- 9391517 TI - Human behaviour and cultural context in disease control. PMID- 9391518 TI - The relevance of anthropology for tropical public health: a historical perspective. AB - Despite the fact that tropical diseases have long been linked to human behaviour and local cultures, the written record of anthropologists involved in the study of tropical disease is relatively recent. In this article, the authors review how anthropologists have shifted their focus from the ecological modeling of disease transmission to the more recent cultural-epidemiological and historical approaches. The list of authors reviewed does not pretend to be exhaustive but rather is intended to reveal the co-existence of multiple paradigms in explaining the emergence of tropical disease within cultures. PMID- 9391519 TI - What anthropology can contribute to tropical medicine: overview of methods for control programmes. AB - In the last two decades anthropologists have begun to carry out studies in tropical medicine and health scientists have followed with the use of qualitative methods and techniques that represent a more person-centered view of planning control programmes. This paper is an overview of selected works by anthropologists and others that suggest the effectiveness of anthropological methodology. PMID- 9391520 TI - Anthropological contributions to a community-based schistosomiasis control project in northern Cameroun. AB - This paper describes how anthropological contributions and extensive cooperation between tropical medicine and medical anthropology researchers contributed to a successful community-based cost recovery schistosomiasis control project in northern Cameroun. The project led to increased knowledge about urinary schistosomiasis by local people, significant decreases in prevalence and intensity of the disease, and increased utilization of primary health care centers. PMID- 9391521 TI - Factors affecting knowledge of the symptoms of schistosomiasis in two rural areas near Ismailia, Egypt. AB - The primary method of control of schistosomiasis in Egypt is through passive chemotherapy, in which people who suspect they have the disease are encouraged to go to their local health unit to be tested and treated. If people are unable to recognize the symptoms of schistosomiasis, this strategy may fail. This paper presents data on local knowledge of the symptoms of schistosomiasis from two areas recently reclaimed from the desert near Ismailia. Using data from free listing and triadic comparisons, it is shown that schistosomiasis is primarily seen as a urinary disease. Factor analysis performed on a series of 12 questions on the symptoms of schistosomiasis included in a survey demonstrated that responses group into three patterns, the first stressing constitutional symptoms such as weakness, the second stressing abdominal symptoms and the third blood in the urine, burning on urination and blood in the stool. The paper discusses the implications of these findings for efforts to promote regular treatment with praziquantel of people living in or near the Nile Delta who are at risk for intestinal schistosomiasis. PMID- 9391522 TI - Human behaviour, cost-effectiveness analysis and research and development priorities: the case of a schistosomiasis vaccine. AB - Cost-effectiveness analysis has been widely used in the health sector to guide decisions about where scarce resources aimed at disease prevention or control should be invested. It has rarely been used to guide decisions about what type of health research should be funded. In addition, the validity of the behavioural assumptions underlying the economic analysis is rarely considered explicitly. This paper explores the use of cost-effectiveness analysis to set priorities for research using the development of a schistosomiasis vaccine as an example. It then explicitly considers behavioural factors which might affect the accuracy of the calculations. A 'product profile' for the new technology is derived which can be used by developers as a target to aim at. To ensure that the vaccine would be more cost-effective than the currently preferred option for the control of schistosomiasis, chemotherapy based on praziquantel, researchers need a vaccine which has sufficient duration of protection to be delivered as part of the regular childhood immunization programme me. The cost of adding it to existing vaccination schedules should not be more than US$4.30 per child in excess of the cost of one round of chemotherapy. It should, ideally, have an efficacy over 80%. These results, however, depend on a number of cultural and behavioural factors which are often ignored in cost-effectiveness studies. For example, low rates of school attendance would increase the cost of contacting children for a chemotherapy programme and increase the relative attractiveness of a vaccine. For chemotherapy to be effective, children also need to comply each year for a number of years. Falling rates of compliance over time would reduce the effectiveness of chemotherapy and increase the attractiveness of a vaccine. But on the other hand, even though a vaccine may still be more cost-effective than chemotherapy at relatively low levels of vaccine efficacy, if mothers perceived the vaccine to be ineffective and refused to bring their children for vaccination, the success of the entire childhood immunization programme could be threatened. PMID- 9391523 TI - Historical perspective of the use of bowel in urology. AB - The use of bowel has been used in urinary tract reconstruction for more than a century. In the past 20 years, however, indications and methods for bowel utilization have multiplied enormously. This article outlines some of these exciting developments. PMID- 9391524 TI - The use of in bowel urology. Metabolic and nutritional complications. AB - Metabolic and nutritional complications of urinary diversion through bowel or stomach segments are common, but fortunately, not often severe. When metabolic abnormalities are problematic, deterioration or baseline insufficiency in renal function is the most likely cause. Deterioration is most commonly associated with obstruction or infection. The urologist should be acutely aware of the potential for metabolic derangements when the prediversion creatinine is greater than 2.0 mg/dL. In this situation, the urologist should employ the basic principles in this article when planning the procedure in order to minimize metabolic complications and morbidities. In the setting of significant renal insufficiency, a short colon or ileal conduit would likely be superior to an ileal or colonic neobladder, or a diversion, incorporating a large gastric segment. Furthermore, in the absence of symptomatic metabolic abnormalities, we advocate treatment of minor laboratory abnormalities, particularly acidosis, to reduce the incidence of metabolic bone disease. Nutritional and gastrointestinal complications are treated on an "as needed" basis, with the exception of metabolic bone disease, which we would hope to prevent with alkalinization and Vitamin C supplementation. Some of the nutritional and gastrointestinal complications are best avoided by leaving the ileocecal valve intact, or by minimizing the use of certain segments. Some evidence exists that over time, histologic changes in the epithelium of diversion segments may impair absorption and contribute to greater resistance against metabolic derangements. Whether the changes truly reduce the incidence of metabolic abnormalities remains to be studied. The ideal, complication-free, diversion with universal application does not exist; however, the urologist must strive to select an option that will provide a functional result for the patient with minimal associated morbidity. PMID- 9391525 TI - Carcinogenesis. The fate of intestinal segments used in urinary reconstruction. AB - The actual mechanism for risk of developing cancer in intestinal segments used for urinary diversion remains uncertain. The clinical and laboratory experiences are reviewed in this article. The pathogenesis is multifaceted, involving initiators and promoters of carcinogenesis. Molecular genetic technology may provide the key to decoding the mechanisms involved. PMID- 9391526 TI - Stomal construction, complications, and reconstruction. AB - This article reviews stomal complications and their management. To accomplish this goal, the authors review techniques used for planning and creating the stomas for both continent reservoirs and incontinent conduits. PMID- 9391527 TI - Noncontinent urinary diversion. AB - When the need for urinary diversion arises, whether from carcinoma of the urinary tract, malfunction, or malformation, a decision must be made about the type of diversion to be performed. Currently, the patient and surgeon must decide on continent versus noncontinent versus neobladder, and on the type of intestinal segment to be used. PMID- 9391528 TI - Augmentation cystoplasty. AB - The past 15 years have been witness to an explosion in the number of reconstructive procedures using bowel in the urinary tract. As with many concepts in medicine, one must rely on clinical experience while laboratory models and other advancements develop. This article attempts to address bladder reconstruction by enterocystoplasty, as well as the indications for augmentation, types of procedures available, and the early and late complications. PMID- 9391529 TI - The Kock pouch urinary reservoir. AB - The use of detubularized ileum for the Kock pouch produced a low pressure, high capacity system superior to large bowel segments that provided excellent continence and protection of the upper urinary tracts. The early enthusiasm was tempered, however, by the technically demanding aspects of the construction of the nipple valves, the early and late complications, and occasional catheterization problems. With simple modifications in the fixation of the intussuscepted nipples, limiting use of staples and mesh collars, and tapering of the stoma, much of those problems have been resolved. The nipple valve is reliable and superior to the tunneled implant for the dilated ureter. With more widespread indications for continent neobladders, the hemi-Kock reservoir remains one of the most dependable and stable neobladders. PMID- 9391530 TI - The Indiana pouch continent urinary reservoir. AB - The right colon reservoir using a stapled plicated ileal efferent limb (Indiana continent urinary reservoir) has been demonstrated to be a reproducible durable form of continent diversion. The overall day and nocturnal continence rate of 94% compares favorably with all other forms of continent cutaneous diversion. Carefully following the technique of stapling and plicating the ileal efferent limb and ileocecal valve as described in this article nearly ensures adequate competence of the outlet valve. In the rare case in which incontinence occurs, it is almost always on the basis of high-pressure unit contractions of the reservoir. On occasion, patients who develop incontinence are observed to have high pressures within the reservoir despite complete detubularization of the right colon segment. When this problem is encountered it can be corrected successfully by adding an ileal patch augmentation to the previously detubularized reservoir. The issue of ureteral implantation in continent urinary diversions is as yet unsettled. Many authors have not used ureteral tenial tunnels and have reported a reflux rate of < 13%. Furthermore, these patients have not developed any long-term sequelae of their reflux. Although favorable results have been obtained without creating tunneled tenial reimplantation, we believe that continent cutaneous reservoirs are almost always colonized with bacteria, and an antireflux mechanism may offer protection against subsequent pyelonephritis. Closure of the reservoir traditionally has been conducted by hand at our institution; however, the development of smaller absorbable gastrointestinal anastomosis stapling devices offers the theoretic advantage of shortening the operative time. We anxiously await follow-up, including larger patient numbers and longer term follow-up of the absorbable staple technique. The use of continent cutaneous urinary diversion clearly has decreased as bladder replacement has become a more viable procedure over the past decade. Despite this, the urologic reconstructive surgeon must maintain the ability to perform continent cutaneous diversion in patients who are unwilling to accept the potential for nocturnal incontinence observed in all forms of bladder replacement as well as the patients who have ineffective sphincter mechanism or who need a urethrectomy due to their primary disease. PMID- 9391531 TI - Ileal orthotopic bladder substitutes. What we have learned from 12 years' experience with 200 patients. AB - The perfect bladder substitute has not been devised yet. The ileal orthotopic bladder substitute, however, provides adequate capacity, convenient voiding patterns, optimal continence rate, preservation of renal function, acid-base balance, and calcium metabolism. The authors describe important surgical details based on experience with more than 200 patients. To achieve a good functional result, patient selection, postoperative voiding reeducation, and meticulous follow-up are important. PMID- 9391532 TI - The Mitrofanoff principle in continent urinary reconstruction. AB - Continent urinary diversion has increasingly become important for treating children and adults with urinary tract pathology that cannot be managed by direct reconstructive techniques. The Mitrofanoff principle, a term that has become synonymous with the flap valve mechanism for promoting the unidirectorial flow of a fluid medium, is a recapitulation of nature's design for the competent ureterovesical junction. Construction of a catheterizable channel using this principle can be performed with a variety of tissues and serves well as a continence mechanism for either the native bladder or intestinal reservoirs. In addition to its utility in managing urinary incontinence, implantation of a catheterizable channel into the cecum can be used to manage fecal incontinence in patients with neurogenic bowel dysfunction. PMID- 9391533 TI - Ileal ureter. AB - Ureteral replacement with ileal bowel segments has become common in the armamentarium of the reconstructive urologist. The use of ileal bowel substitution, whether total or segmental, has provided yet another surgical alternative for renal preservation. The indications, surgical technique, and results with the ileal ureter are reviewed. PMID- 9391534 TI - The ileal neobladder to the female urethra. AB - In the author's opinion, in the properly selected woman undergoing radical cystectomy for transitional cell carcinoma of the bladder, the ileal neobladder to the female urethra is a viable option. Ten years of experience with 23 patients have led to a nerve and urethral support cystectomy technique with the ileal neobladder anastomosed to the proximal urethra. Even then, however, retention in 20% of patients rather than the expected incontinence is the critical issue. Incontinence has never been a problem. The advent of orthotopic lower urinary reconstruction in women is a major achievement in the evolution of urinary diversion. With our increasing understanding of the continence mechanism in women and with increasing evidence that the female urethra can be safely preserved after cystectomy, orthotopic lower urinary tract reconstruction by the ileal neobladder can now be offered safely not only to men but also to women undergoing cystectomy with superb functional results. PMID- 9391535 TI - Use of small and large bowel in renal transplantation. AB - The continued success of renal transplantation has provided a higher quality of life for properly selected patients with ESRD. It is also a much more cost effective and efficient treatment of ESRD compared with chronic dialysis. Innovative urologic reconstructive surgery using enteric segments for both continent and incontinent urinary diversions has permitted this therapeutic modality to be offered to the recipient with lower urinary tract disease not previously amenable to renal transplantation. These same reconstructive techniques using ileal segments have also permitted preservation of renal allografts with previously nonreconstructable renal pelvic or ureteral disease. PMID- 9391536 TI - Bilateral breast carcinoma versus unilateral disease. Review of 498 patients. AB - In literature data, an uncertainty exists whether occurrence of bilateral breast cancer decreases the survival probability of affected patients. Therefore, we analyzed the medical records of 498 postoperatively irradiated (1977-1982) female breast cancer patients (T1-4,N0-3,M0). In the follow-up time, in 36 patients a bilateral breast carcinoma treated by surgery with or without radiotherapy was found. The 10-year overall survival rates were 54% in patients who had unilateral disease, compared with 56% in bilateral carcinoma patients, respectively. The incidence of metastasis did not differ between both groups: 24.2% versus 38.8%. Eleven percent of unilateral cancers recurred; in the other group, local failure of the first and second tumor was observed in 19.4% and 11.1%, respectively. We conclude that the occurrence of bilateral breast cancer has no significant impact on survival, although the development of local failures and metastases seems to be more frequent. The therapeutic strategy in bilateral carcinoma should resemble the treatment procedure in unilaterally affected patients. PMID- 9391537 TI - The prognostic significance of steroid receptor activity in tumor tissues of patients with primary breast cancer. AB - The prognostic significance of steroid-receptor activity is still debatable. Discrepancies in results are probably attributable to few patients, heterogeneous patient populations, and short follow-up. We investigated the prognostic significance of estrogen- and progesterone-receptor (ER and PgR, respectively) activity as a continuous variable in a homogeneous patient population. The prognostic significance of steroid-receptor activity was examined in 329 node negative and 320 node-positive unselected breast cancer patients. In node negative patients, ER values of primary tumors between 100 and 400 fmol/mg protein appeared to be a significant predictor for low risk of recurrence, whereas high ER (> 400) revealed an unfavorable prognosis. The classic cutoff level of ER (< 10 fmol/mg proteins) had no prognostic significance, however. In patients receiving adjuvant chemotherapy--the node-positive breast cancer patients--the classic cutoff value of ER (10 fmol/mg protein) predicts significantly distant metastases-free survival and overall survival only in the first 4 years of follow-up after diagnosis. Progesterone receptor is a time dependent prognosticator in node-negative breast cancer patients (cutoff point for PgR, 80 fmol/mg). In node-positive breast cancer patients treated with chemotherapy or a combination of chemo- and hormonal therapy, PgR values lower than 60 fmol/mg had a worse prognosis. The results show the poor performance of standard cutoff points for ER and PgR positivity in predicting prognosis. Better prognosis is related to higher receptor levels but this relation is predominantly time-dependent. Moreover, patients who have high ER levels have a prognosis that is worse when compared with intermediate ER levels. Standard cutoff points for steroid receptors should not be used to select patients for prognosis. PMID- 9391538 TI - Malignant eccrine spiradenoma. Case report and review of the literature. AB - Malignant eccrine spiradenoma is an exceedingly rare tumor. A case of a 72-year old women with this highly aggressive malignancy arising from a long-standing lower leg lesion is reported. Management during the course of disease included surgery, radiation therapy (RT), hyperthermic limb perfusion chemotherapy, and chemotherapy. The patient died of her disease, with widespread metastatic disease 20 months after the diagnosis. A review of the literature is presented, and treatment considerations are summarized. PMID- 9391539 TI - Use of the somatostatin analogue octreotide acetate in the treatment of encephalopathy associated with carcinoid tumor. Case report. AB - Carcinoid tumors may present in a variety of ways dependent on sites of disease and ectopic hormone secretion. This case report describes a patient having metastatic carcinoid tumor who developed encephalopathy of uncertain etiology. Treatment with the somatostatin analogue octreotide acetate resulted in dramatic improvement in his mental status. Several plausible mechanisms are discussed. PMID- 9391540 TI - Hypofractionated radiotherapy with 5-fluorouracil radiosensitization for locally "far advanced" breast cancer. AB - Seventeen patients who had locally far-advanced breast cancer were treated with hypofractionated radiotherapy (4-5 Gy/fraction, twice a week) and concomitant 5 fluorouracil (5-FU 300 mg/m2 intravenously, 1 hour before every radiotherapy fraction). Fourteen of the seventeen patients had disease that was not responding to chemotherapy. Early toxicity was low and none developed grade III/IV toxicity. Two of the seventeen patients showed moist skin desquamation and four of seventeen had grade II anemia. Of eight patients who survived longer than 12 months, symptomatic breast fibrosis was observed in one (12%), asymptomatic pericarditis in one (12%) and symptomatic radiation pneumonitis in one (12%). Plexopathy and arm edema grade II were observed in one patient and two patients, respectively. Quality of life substantially improved. Complete response was documented in five of the seventeen patients (29%), with pathologic confirmation in three. Seven of the seventeen (41%) patients were considered to be partial responders, four (23%) had a minimal response, and one (6%) progressed during treatment. Local progression-free survival (1-24 months) was achieved in 12 of 17 patients. Four of the seventeen (23%) patients are alive, with no evidence of disease (local or distant) 8 to 24 months after radiotherapy. Hypofractionated chemoradiotherapy with 5-FU is an effective, convenient, and well-tolerated regimen for far-advanced breast tumors. PMID- 9391541 TI - Herpetic geometric glossitis in a pediatric patient with acute myelogenous leukemia. AB - Herpetic geometric glossitis, a recently described form of lingual herpes simplex virus type 1 (HSV-1) infection, has been reported in 6 human immunodeficiency virus (HIV) patients and 1 cardiac transplant patient who was receiving immunosuppressant therapy. An HIV-seronegative immunocompromised pediatric patient with acute myelogenous leukemia who developed herpetic geometric glossitis is described. Herpetic geometric glossitis can present in both adult and pediatric immunocompromised patients. The symptoms, morphology, laboratory findings and treatment of this infection are summarized. The possible consequences of untreated herpetic glossitis include superinfection and undernourishment. Although previously described patients responded to 1000 mg per day (divided in 5 doses) or oral acyclovir, with complete resolution of fissures, this patient developed herpetic geometric glossitis while receiving acyclovir and required higher doses of oral antiviral therapy (acyclovir, 3000 mg/day divided in 5 doses) to treat his HSV-1 lingual infection. Empiric treatment of an immunocompromised patient who has newly acquired painful tongue fissures or furrows with systemic acyclovir should be considered. PMID- 9391542 TI - A comparative study of intravenous granisetron versus intravenous and oral ondansetron in the prevention of nausea and vomiting associated with moderately emetogenic chemotherapy. AB - We conducted a prospective, randomized, open, single-center, parallel group study comparing the anti-emetic efficacy and toxicity of granisetron with that of ondansetron in patients receiving moderately emetogenic chemotherapy. From December 1994 to May 1995, patients who were to receive moderately emetogenic chemotherapy for the first time or who had not received chemotherapy (80 to 100 mg/m2 of cisplatin or 40 mg/m2 of doxorubicin) within 4 weeks previously were enrolled in this study. The following anti-emetic regimens were used: 3 mg of granisetron were given intravenously before chemotherapy for a single dose; 8 mg of ondansetron were given intravenously before chemotherapy and then every 8 hours for a total of 3 doses, plus 8 mg of an oral maintenance dose every 12 hours for 5 consecutive days. We evaluated 97 patients (48 received granisetron and 49 received ondansetron). In the first 24 hours after chemotherapy, complete and major responses were achieved in 76.6% of the patients receiving granisetron and in 72.9% of patients receiving ondansetron (p = 0.9033). Additionally, there was no difference in the control of delayed nausea and vomiting between the two groups (51.1% versus 54.2%, p = 0.9200), and there were no significant adverse effects or toxicities. We have concluded that a single dose of granisetron is as effective in prophylaxis of emesis induced by moderately emetogenic chemotherapy as a triple dose of ondansetron plus oral maintenance. PMID- 9391543 TI - Phase II trial of intravenous CI-980 (NSC 370147) in patients with metastatic colorectal carcinoma. Model for prospective evaluation of neurotoxicity. AB - CI-980 (NSC 370147)--a synthetic mitotic inhibitor that binds to tubulin at the colchicine binding site--has significant activity against a broad spectrum of tumor models and greater in vitro cytotoxicity when given over > 24 hours than 4 hours or less. Phase I studies demonstrated central nervous system (CNS) toxicity to be dose-limiting when CI-980 was administered as a 24-hour infusion. When a 72 hour infusion was given, CNS toxicity was reduced and granulocytopenia became the dose-limiting toxicity. In this phase II study, CI-980, 4.5 mg/m2, was administered as a 24-hour continuous intravenous infusion for 3 consecutive days and repeated every 21 days. Fourteen patients who had measurable metastatic colorectal cancer were entered in the trial. Eight patients had received one prior chemotherapy regimen for metastatic disease. Patients were prospectively monitored by neurologic examinations and neuropsychologic assessment of cognitive functioning. No complete or partial responses were observed. Grade 4 granulocytopenia was the dose-limiting toxicity. Reversible declines in recent memory function were noted in all patients. After each course of CI-980, there were also transient non-significant declines in motor coordination, compared with the preinfusion assessment. At the stated dose and schedule, CI-980 lacks activity in metastatic colorectal carcinoma. The agent's toxicity profile (granulocytopenia and CNS effects) was comparable with previously described effects of this agent. PMID- 9391544 TI - Secondary syringomyelia due to intramedullary spinal cord metastasis. Case report and review of literature. AB - Intramedullary spinal cord metastasis is relatively rare. We describe a patient having intramedullary spinal cord metastasis associated with syringomyelia, confirmed by magnetic resonance imaging, in a patient who had poorly differentiated carcinoma of the lung. The patient responded to treatment with steroids and radiotherapy, with complete resolution of neurologic symptoms and syringomyelia. PMID- 9391545 TI - Neoadjuvant chemotherapy followed by concurrent chemotherapy and radiotherapy for locally advanced esophageal carcinoma with bulky upper abdominal lymphadenopathy. Case report. AB - A 60-year old male patient who had locally advanced esophageal carcinoma with bulky upper abdominal lymphadenopathy underwent neoadjuvant chemotherapy consisting of 5-fluorouracil (5-FU) and cisplatin (CDDP), followed by concurrent radiotherapy and chemotherapy using protracted low-dose continuous infusion of 5 FU and CDDP. The treatment brought about complete remission in the primary lesion and good partial remission in the upper abdominal lymphadenopathy. He subsequently underwent trans-hiatal esophagectomy after one cycle of adjuvant chemotherapy because local recurrence was suspected. Histopathologic study of the resected specimen demonstrated no malignant tissue in the primary lesion and the lymph nodes. The patient is still alive and disease-free at 26+ months. This result suggests that neoadjuvant chemotherapy followed by concomitant chemotherapy and radiotherapy for patients who have locally advanced squamous cell carcinoma of the esophagus with intensive abdominal lymphadenopathy may offer some chance for sterilization of local and regional metastases and longer survival. PMID- 9391546 TI - The heterogeneity of leukemia occurring after treatment for sarcoma. AB - The successful treatment of sarcomas with intensive regimens combining high-dose chemotherapy and irradiation has led to not only improved survival but also to an increased incidence of therapy-related acute non-lymphocytic leukemia (t-ANLL) and myelodysplastic syndrome (MDS). We report 4 patients having sarcoma treated with chemotherapy or chemoradiotherapy who subsequently developed MDS or t-ANLL. PMID- 9391547 TI - Contamination of the pleural surfaces in childhood sarcoma. Use of colloidal P-32 to reduce radiation dose to the whole lung. AB - Children with pulmonary sarcomas who have diffuse contamination of the pleural cavity present a difficult management problem for the radiation oncologist. Doses required to control even microscopic disease exceed lung tolerance. We report on the use of intracavity colloid P-32 in an attempt to treat the pleural surface and spare normal lung parenchyma and tissues of the chest wall. Three children- 18 months, 12 years, and 3 years of age--had spillage of pulmonary sarcomas into the chest cavity. All children were treated with systemic chemotherapy. Initially, 0.5 mCi of technetium sulfur colloid (99mTc-sulfur colloid) was instilled into the pleural space to ascertain even distribution of isotope. This was then followed by installation of 5.0 mCi of colloidal P-32. Uniform distribution was then confirmed by bremsstrahlung scanning. All three patients are in complete remission 3.5 years, 3 years, and 1 year after treatment, respectively. The major toxicity was asymptomatic pleural thickening, which could be confused with disease. This was confirmed histologically to be fibrous in the first patient. The process diminished or stabilized with time in all 3 patients over the period of observation. In this small series, intrapleural colloidal P-32 appeared to be safe and well tolerated and would be expected to be less toxic than wide-field external beam in the treatment of spilled pulmonary sarcomas. PMID- 9391548 TI - Predicting drug interactions in vivo from experiments in vitro. Human studies with paclitaxel and ketoconazole. AB - This study was performed to evaluate whether concomitant treatment with ketoconazole could reduce the clearance of paclitaxel given to ovarian cancer patients. Paclitaxel, 175 mg/m2, was given as a 3-hour continuous intravenous infusion and repeated every 21 days. Initially, ketoconazole, 100 to 1600 mg, was given as a single oral dose 3 hours after paclitaxel. Later, ketoconazole, 200 mg, was given perorally 3 hours before paclitaxel. Plasma drug concentrations were measured by high-pressure liquid chromatography (HPLC), and cytochrome P450 3A (CYP3A) activity was measured with the erythromycin breath test (ERMBT). Ketoconazole did not alter plasma concentrations of paclitaxel or its principal metabolite, 6 alpha-hydroxypaclitaxel. Although there was marked inter- and intrapatient variability in ketoconazole pharmacokinetics, peak plasma concentrations in all but one course were below the 50% inhibitory concentration (IC50) point determined for inhibition of paclitaxel metabolism in vitro. Therefore, paclitaxel and ketoconazole can be coadministered safely without dose adjustments. There was no correlation between ERMBT measurements and serial plasma concentrations of paclitaxel. The erythromycin breath-test measurements did correlate with the corresponding ketoconazole plasma concentrations. The erythromycin breath test is a valuable tool for measuring instantaneous CYP3A activity in vivo. This clinical study confirms the results of prior studies with human-derived materials in vitro, reinforcing the notion that such studies are useful predictors of drug pharmacokinetics and interactions in vivo. PMID- 9391549 TI - Comparison of patient characteristics and outcome between a single-institution phase II trial and a cooperative-group phase II trial with identical eligibility in metastatic melanoma. AB - Differences in overall survival and response rates are often noted when promising single-institution phase II treatment regimens are evaluated in a cooperative group setting. One reason for this discrepancy may be the differences in patient characteristics at the time of registration. In the metastatic melanoma literature, the prognostic factors for survival that are most frequently identified are the number of metastatic sites, visceral involvement, performance status, liver involvement, and possibly the disease-free interval and gender. A prognostic factor for response appears to be sites of involvement. A comparison of patient characteristics and outcome was conducted for patients entered in similar phase II melanoma trials at a single institution versus those in a cooperative group. Sixty-four patients at Wayne State University (WSU) were compared with 96 patients who had nearly identical eligibility criteria and were registered in the Southwest Oncology Group (SWOG). All patients were receiving comparable phase II treatments for metastatic melanoma. Southwest Oncology Group patients were significantly older (p < 0.001), had worse performance status (p = 0.03), had more visceral involvement (p = 0.001), and were more likely to have two or more metastatic sites (p = 0.02). No significant differences in gender (p = 0.55), absence or presence of liver involvement (p = 0.12), or disease-free interval were noted. These disparities, despite similar eligibility, may partly explain the observed differences in survival (WSU median = 10 months, SWOG median = 7 months; p = 0.13) and response rates (WSU = 31%, SWOG = 15%; p = 0.02) between the two groups of patients. Investigators should report these important patient characteristics in treatment reports. These differences highlight the difficulty in comparing single-institution and cooperative-group phase II trials, even with comparable patient eligibility. This serves to emphasize the need for well-designed phase III trials when comparing treatment approaches and stratification for the prognostic factors identified. PMID- 9391550 TI - Low serum testosterone and a younger age predict for a poor outcome in metastatic prostate cancer. AB - Carcinoma of the prostate gland is one of the most common malignancies in males. This study was undertaken to determine which factors predict the course and outcome of patients treated with first line hormonal manipulation. A total of 144 patients with Stage D2 prostate cancer who received androgen deprivation therapy were studied. Pretreatment parameters analyzed were age, performance status, analgesia usage, concurrent disease, histologic differentiation, hemoglobin, leukocyte and platelet count, serum creatinine, alkaline phosphatase, lactate dehydrogenase, prostate specific antigen, total and prostatic acid phosphatase, serum testosterone, follicle stimulating and luteinizing hormone levels, number of metastatic sites and bone scan grade. Only initial serum testosterone (> 10 nmol/l) had a positive impact on response (p = 0.0304), whereas age older than 60 years had a positive impact on time to progression (16 vs. 11 months, p = 0.0414). Both serum testosterone (26 vs. 20 months, p = 0.003), and age (28 vs. 17 months, p = 0.036) had a significant influence on overall survival. Low testosterone, indicating androgen independence, and a younger age, seem to result in a more aggressive disease and a poorer prognosis in advanced prostate cancer. PMID- 9391551 TI - Safety of radiating jejunal interposition grafts in head and neck cancer. AB - The safety of high-dose postoperative radiation therapy to a jejunal graft has not been established in the literature. The purpose of the present study is to review the effect of postoperative radiation on swallow function in patients who have received a jejunal interposition graft as part of their reconstruction after cancer resection. Charts of patients undergoing hypopharyngeal resections for cancers with placement of jejunal interposition grafts who received postoperative radiation therapy were reviewed. Swallow function was determined from records of patients' subjective characterization of their swallow function, records of weights at each visit, use of gastrostomy tube, need for jejunal dilatation and review of barium swallows. Seventeen patients were identified who had undergone resection of cancers with jejunal interpositions and postoperative radiation therapy. Four patients never regained adequate swallow function postoperatively to allow G-tube removal. The remaining thirteen patients had their G-tubes removed, generally several months after resection, and were able to obtain adequate nutrition orally to maintain or increase their weights. The present series suggests that a segment of jejunum transferred into the neck after laryngopharyngoesophagectomy can be irradiated to high dose with generally acceptable morbidity. PMID- 9391552 TI - Continuous infusion of carboplatin during conventional radiotherapy treatment in advanced squamous carcinoma of the cervix uteri IIB-IIIB (UICC). A phase I/II and pharmacokinetic study. AB - OBJECTIVE: A prospective, single-arm phase-I/II trial performed to assess the efficacy and toxicity of the concomitant use continuous infusion of low-dose carboplatin and pelvic conventional radiotherapy in patients with locally advanced squamous cell carcinoma of the cervix. MATERIALS AND METHODS: Between January and July 1994, a total of 12 patients consecutively diagnosed to have squamous cell carcinoma of the cervix uteri stages IIB-IIIB UICC-TNM (five patients, IIB; and seven patients, IIIB) entered the study. All patients were evaluated by a gynecologist and a radiation oncologist and were submitted to standard pretreatment staging procedures. Radiation was delivered with 10-MeV photon beams with the shrinking-field technique. The patients received 2 Gy radiotherapy daily, 5 fractions per week, up to a planned total of 60 Gy in 6 weeks to the primary tumor and 46 Gy in 4 weeks to the whole pelvis. Irradiation was performed using four fixed orthogonal fields. One intracavitary insertion, 8 Gy to point A (dose rate, 1.1 Gy/h), was performed immediately after external pelvic irradiation. Carboplatin (12 mg/m2/day) was also administered in a continuous infusion, starting 1 day before the first fraction of radiotherapy. The platinum in plasma and urine, as well as the platinum concentration in the cytosols of lymphocytes and tumor, was measured weekly. RESULTS: A complete response was seen in nine (75%) of the 12 patients. Of the nine patients who achieved a complete remission, only one had subsequent failure in the pelvis. The total pelvic failure rate was 33.3% (four of 12 patients). With a median follow up time of 20 months, the actuarial survival rate at 24 months was 64.8%. All patients completed the treatment without major protocol violations. Grade-2 leukopenia (in nine patients) and grade-1 nausea and vomiting (in five) were the most common acute toxicities. There was one grade-3 hematologic toxicity. Grade-3 late complications were observed in 16.6% of cases (two of 12 patients). On days 28 and 42 of the treatment, the mean total platinum plasma concentrations were 491 micrograms/L (SD = 129) and 672 micrograms/L (SD = 160), and the ultrafilterable fraction was 8-10%. At the same time points, the concentration in lymphocytes was constant at 21 picograms (pg) platinum/lymphocyte. The levels of platinum concentration measured on days 14 and 28 in the cytosols of tumor cells were 0.3 microgram/g (SD = 0.1) and 0.93 microgram/g (SD = 0.2). CONCLUSION: The combination of continuous infusion of carboplatin and radiotherapy at the aforementioned doses in patients with locally advanced cervical carcinoma resulted in a relatively low frequency of significant acute and late complications. Platinum in normal tissue (picograms per lymphocyte) was stable from week 1 of treatment, whereas the platinum steady state in plasma and in tumor cells was not reached in 6 weeks and was below that required in vitro to produce radiopotentiation. Further studies to determine the optimal dose of carboplatin and irradiation are needed prior to the initiation of phase-III studies. PMID- 9391553 TI - A phase II study of topotecan administered five times daily in patients with advanced gastric cancer. AB - OBJECTIVE: Topotecan (Tpt), a semisynthetic analogue of camptothecin (Cpt), has shown excellent preclinical activity in a number of solid tumors. Cpt, the parent compound, has preclinical activity against several gastrointestinal tumors, including a gastric adenocarcinoma xenograft. A phase-II clinical trial was conducted to assess the activity to Tpt in patients with advanced gastric cancer. MATERIALS AND METHODS: 15 patients with advanced, incurable gastric adenocarcinomas were treated. Tpt 1.5 (mg/m2/day) was administered intravenously as a 30-min infusion daily for 5 consecutive days. Treatments were repeated on a 21-days cycle. RESULTS: No major objective responses were observed in 13 evaluable patients (response rate = 0%; 95% confidence interval = 0-22%). The major dose-limiting toxicity in this trial was myelosuppression, which was severe in this patient population. CONCLUSIONS: Tpt at the dose and schedule studies does not possess substantial antitumor activity in patient with gastric cancer, and the toxicities were formidable. We do not advocate further development of this drug in the treatment of gastric cancer. Tpt has shown more promising activity and tolerability in other patient populations, and these areas deserve further exploration. PMID- 9391555 TI - A rare event of megestrol acetate (Megace)-induced adrenal suppression in a breast cancer patient. PMID- 9391554 TI - A phase II trial of amonafide in patients with nonsquamous cell carcinoma of the cervix. A Gynecologic Oncology Group study. AB - Twenty-seven patients with nonsquamous cell carcinoma of the cervix were entered into a Phase II study of amonatide; 24 patients were evaluable for toxicity, while 23 were evaluable for response. Patients received amonafide, 300 mg/m2, intravenously for 5 consecutive days every 3 weeks. The median age of patients was 45 years. All but two patients were completely ambulatory. Twelve patients had received prior chemotherapy, while 22 had been treated with radiation therapy. One of 27 (4.3%) patients had a partial response (PR) to this regimen and 13 (56.5%) had stable disease. Sixteen patients experienced a median white blood cell (WBC) nadir of 350/mm3, seven developed life-threatening thrombocytopenia, and one had severe anemia requiring transfusion. Nonhematologic toxicity was mild. Amonafide had insignificant activity in these patients with nonsquamous cell carcinoma of the cervix. PMID- 9391556 TI - Advances in skin cancer. A tribute to Professor Frederick Helm on the occasion of his 70th birthday. PMID- 9391557 TI - The actinic keratosis. A perspective and update. AB - BACKGROUND: Actinic keratosis (AK) is a common sun-induced precancerous neoplasm confined to the epidermis. It is the initial manifestation of a continuum of clinical and histologic abnormalities that progresses to invasive squamous cell carcinoma (SCC), a disorder that accounts for thousands of preventable deaths in America each year. OBJECTIVE: The purpose of this work is to describe the actinic keratosis. METHODS: This effort was performed by a literature review and analysis. RESULTS: Like SCCs, the vast majority of AKs are asymptomatic. Although some actinic keratoses may become clinically inapparent, possibly either due to immune rejection or simply having their external surface unknowingly scraped off, an untreated AK represents a potentially curable fatal cancer. CONCLUSIONS: Each AK should be treated before it progresses to invasive squamous cell carcinoma. Destructive modalities such as cryosurgery using liquid nitrogen and electrodesiccation and curettage are the mainstays of therapy. Each case must be individualized. LEARNING OBJECTIVES: After studying this article, participant should be able to: 1. Understand the concept of an actinic keratosis. 2. Learn how to recognize its clinical manifestations. 3. Be aware of the danger it poses as an easily curable papulonodule that may become a fatal cancer. PMID- 9391558 TI - Presence of human papillomavirus DNA and expression of L-fucose moiety in some vulvar intraepithelial lesions and vulvar squamous cell carcinoma. AB - BACKGROUND: Human papillomavirus could reside and play an etiological role in some vulvar epithelial lesions. Human papillomavirus-infected keratinocytes might have certain biochemical changes that could be of significance in helping clinical diagnosis or elucidating the pathogenesis of some vulvar epithelial diseases. OBJECTIVE: To detect the presence of human papillomavirus DNA and observe the expression pattern of L-fucose moiety in some vulvar intraepithelial lesions and vulvar squamous cell carcinoma. METHODS: Nineteen cases of vulvar intraepithelial lesions and 13 cases of vulvar squamous cell carcinoma from the inner aspect of labia majora or the outer aspect of labia minora were selected. Polymerase chain reaction was employed to detect the presence of human papillomavirus DNA in lesional skin. Ulex europeaus agglutinin-1 histochemistry was used to observe the L-fucose expression pattern. RESULTS: A large proportion of vulvar intraepithelial lesions had the presence of human papillomavirus DNA, but the positive rate was low in vulvar squamous cell carcinoma and squamous cell hyperplasia. L-Fucose expression was much more pronounced in human papillomavirus DNA-positive lesions than those negative ones. CONCLUSIONS: Human papillomavirus had an important impact on some vulvar intraepithelial lesions in this Oriental population. The expression pattern of L-fucose may be used as an indicator for vulvar keratinocyte transformation by human papillomavirus. PMID- 9391559 TI - Perianal Paget's disease years after rectal adenocarcinoma removal. AB - BACKGROUND: Perianal Paget's disease often coincides with anorectal carcinoma, which extends into the epidermis from a contiguous organ. OBJECTIVE: Our purpose was to present a patient with perianal Paget's disease who had a rectal adenocarcinoma excised 14 years previously in another hospital and to determine whether there is a relationship between the perianal Paget's disease and the rectal adenocarcinoma. METHODS: We examined the resected specimens of the rectal adenocarcinoma and the perianal Paget's disease histologically. RESULTS: In the resected specimen of the rectal adenocarcinoma, Paget cells were present within the anal epidermis adjacent to the rectal adenocarcinoma. The Paget cells showed the same histochemical and immunohistological findings as the adenocarcinoma cells. CONCLUSION: There was a close relationship between the perianal Paget's disease and the rectal adenocarcinoma. It is probable that the Paget cells were derived from direct spread from the rectal adenocarcinoma. PMID- 9391560 TI - Triple extramammary Paget's disease. AB - BACKGROUND: Triple extramammary Paget's disease (TEPD) has been considered to be rare in the English literature, and its incidence and characteristics are unclear. There are many therapeutic options for treating extramammary Paget's disease (EPD). OBJECTIVE: Our purpose was to investigate how many TEPD cases have been reported previously and to describe their characteristics. We also describe the effectiveness of radiotherapy for them. METHODS: We report two TEPD cases, and summarize previously reported TEPD cases together with our cases. RESULTS: Twenty-three TEPD cases have been reported previously. Of these, 19 cases have been in Japan. All but one patient with TEPD were male. Their axillary lesions often showed no eruptions or very slight erythema. Radiotherapy for our cases was effective although the effectiveness of radiotherapy is controversial. CONCLUSION: In genital Paget's disease bilateral axillae should be examined histologically, even if they show no or slight eruptions. Radiotherapy may be useful for EPD, particularly axillary Paget's disease. PMID- 9391561 TI - Identification of melanoma risk factors in the Polish population. AB - BACKGROUND: There is a great deal of literature regarding malignant melanoma risk factors all over the world. However, such data concerning the Polish population has not been published as of yet. OBJECTIVE: To identify the importance of melanoma risk factors including number and distribution of common and atypical nevi of the patients, phenotypic features, and environmental conditions. METHODS: This is a case-control study of 74 melanoma patients and 300 controls. We used histopathological examination to confirm diagnosis of melanoma. Epiluminescence microscopy was introduced for differential diagnosis of all pigmented lesions. RESULTS: The mean number of common nevi in melanoma patients was significantly higher compared with the control group (22.1 vs 15.2; P < 0.05). Atypical nevi in the melanoma patients group and in the control group were encountered in 16.2% and 6.6%, respectively (P < 0.05). In the melanoma group more patients with fair complexion, intense sun exposure, and sunburn were identified compared with the control group. CONCLUSION: It was stated that an increased number of nevi (especially atypical ones), fair skin, and blue/green eyes, as well as intense UV exposure and sunburns, are important risk factors for melanoma development. PMID- 9391562 TI - Does surgical removal of primary melanoma trigger growth of occult metastases? An analytical epidemiological approach. AB - BACKGROUND: In several human tumor systems a potential role of surgical removal of the primary tumor upon metastatic tumor growth has been evaluated, as it has been in experimental models. The present study addresses the question of whether the removal of primary melanomas disinhibits growth of metastatic disease and results in more rapid progression. METHODS: In a data set of 1224 primary cutaneous melanomas the risk of "thin" melanomas to develop metastases within 1 year was compared with the risk of matched pairs of "thick" melanomas to present metastases at the time of diagnosis. For this purpose, a pairwise matching procedure based on certain assumptions as to tumor volume and tumor doubling time has been applied. RESULTS: When a long tumor doubling time is assumed (200, 400, or 800 days), the tumors removed seem to have a significantly higher risk of metastases to become clinically apparent within 1 year than the matched pairs of tumors to present metastatic disease at the time of diagnosis (chi-square < 0.01). When short tumor doubling time is assumed (50 or 100 days), the difference is not significant, but there also seems to be no benefit for the operated patients. CONCLUSION: In the present data set there is evidence that surgery of primary melanoma may enhance tumor growth at metastatic sites. PMID- 9391563 TI - Malignant melanomas simulating various types of soft tissue tumors. AB - BACKGROUND: Primary malignant melanomas, recurrences, and metastases thereof can present with a wide variety of clinicopathologic aspects. OBJECTIVE: The present series describes eight primary malignant melanomas and/or 10 local recurrences/metastases (from eight patients) misdiagnosed as various types of soft tissue tumors: two dermatofibrosarcoma protuberans, two atypical fibroxanthomas, two storiform-pleomorphic malignant fibrous histiocytomas, one myxofibrosarcoma ("myxoid malignant fibrous histiocytoma"), two malignant hemangiopericytomas, and nine malignant schwannomas. METHODS: In three cases correct diagnosis was established by clinicopathologic correlation during follow up; all the others were only discovered during a retrospective work-up of all soft tissue tumors diagnosed at the Dermatohistopathological Laboratory of the Department of Dermatology, University of Innsbruck. RESULTS: Helpful clues derived from subtle intraepidermal, irregular spread of melanocytes, focal nested arrangement of tumor cells, sparse melanin deposition, neurotropism, and prominent peripheral lymphohistiocytic response, partially forming lymph follicles. Besides clinicopathologic correlation, histology of serial sections as well as immunohistochemistry (S100 protein more important than NK1/C3; HMB45 without benefit) and electron microscopy (melanosomes) proved helpful for definitive diagnosis. In contrast to the general assumption that spindle cell/desmoplastic malignant melanomas have an unequivocal bad prognosis, our series, as well as evidence from the literature, document a better prognosis than that of other types with comparable tumor thickness (70% vs 50% 5-year survival). Moreover, labeling with E9, an antimetallothionein marker, confirmed its usefulness for prognosis being strongly positive in primary lesions followed by rapid progression, but mostly negative in those without. CONCLUSION: These cases document that malignant melanoma may mimic various types of soft tissue tumors; correct interpretation is important as prognosis and therapeutic management differ considerably between these entities. PMID- 9391564 TI - Cutaneous malignant melanoma treated by Mohs surgery. Review of the treatment results of 179 cases from the Mohs Melanoma Registry. AB - BACKGROUND: There has been much debate regarding the efficacy of the Mohs chemosurgery fixed-tissue technique vs the fresh-tissue technique in the treatment of cutaneous melanoma. OBJECTIVE: To review the treatment results of the 179 cases of melanoma registered with the Mohs melanoma tumor registry from 1981 to 1991, accumulated from nine referring Mohs surgeons. METHODS: Review of the two treatment techniques using a case presentation format. There were 113 cases treated by the hybrid fixed-tissue technique and 61 cases treated by the fresh-tissue technique. The data compared technique of treatment vs degree of invasion by both Clark level and Breslow thickness determinations. Analysis of the data using Kaplan-Meier graph. RESULTS: Five-year survival data for melanomas treated by either the fresh-tissue or fixed-tissue method appear concordant. CONCLUSION: For thin and intermediate melanoma thicknesses treatment by either method appears to be equally efficacious. For deep melanomas, the number of cases were insufficient to evaluate. Further study of this high-risk category is warranted. PMID- 9391565 TI - Tissue repair after Mohs surgery. A plastic surgeon's view. AB - BACKGROUND: Goals of the treatment for skin cancer include completeness of removal of the lesion, minimal functional disability, and a good aesthetic result. With increasing standards for the quality assurance and the demand for cost-effectiveness, assessment of resource-consuming treatment modalities, especially those involving multidisciplinary approaches, seems appropriate. OBJECTIVE: The purpose of this study was to review the strategy of management and the approaches to tissue repair following cutaneous micrographic surgery from the plastic surgeon's point of view. METHOD: Retrospective review of personal experience based on approximately 800 patients treated between 1989 and 1996 and current plastic surgery literature. RESULTS AND CONCLUSIONS: Teamwork with the Mohs surgeon, recognition of the post-Mohs' procedure wound components, and familiarity with reconstructive techniques are essential for the multidisciplinary practice success. The pitfalls of the reconstructive approaches are discussed. PMID- 9391566 TI - Anti-human epithelial antigen (Ber-EP4) helps define basal cell carcinoma masked by inflammation. AB - BACKGROUND: Anti-human epithelial antigen (Dako-Ber-EP4) is an antibody raised in mice that reacts with two glycoproteins of 34 and 49 kD. These glycoproteins are present on the cell surface and in the cytoplasm of basal cell carcinoma (BCC) cells, sweat glands, and some hair follicles in the skin. METHODS: We selected 27 BCCs (15 nodular, 11 morpheic/infiltrative, and one adenoid) and one trichoblastoma and performed rapid immunohistochemical studies with Ber-EP4 and a labeled streptavidin biotin alkaline phosphatase system. RESULTS: Twenty-seven of 27 BCCs and one of one trichoblastoma were positive for Ber-EP4. Thirteen of 27 BCCs stained with Ber-EP4 showed areas of BCC in dense inflammation that were better defined by the Ber-EP4 immunostain than by the H&E stain. In two cases persistent infiltrative BCC was found in the final Mohs margins while appearing negative with routine H&E. Several instances occurred where negative Ber-EP4 in inflammatory fields resulted in tissue sparing with the avoidance of a further Mohs (insurance) layer. CONCLUSION: In conclusion, we found mouse anti-human Ber EP4 a useful and reliable marker for BCC. This antibody helps to locate latent BCC tumor in inflammatory Mohs margins. PMID- 9391567 TI - Combination of an island advancement flap and a composite graft for reconstruction of the nasolabial defect. AB - BACKGROUND: The alar region is one of the most difficult areas of the face to reconstruct. Up until now, various methods have been demonstrated for achieving the best possible results in terms of cosmetic appearance and function. This report deals with a combination of a random pattern flap and a free composite graft, carried out in two stages. OBJECTIVE: In order to reconstruct the alar region, an island advancement flap as well as a composite graft from the contralateral ear were used. METHODS: The defect in the cheek-upper lip region was closed using an island advancement flap. In a second operation 2 weeks later, the reconstruction of the alar region was attempted using a composite graft from the right ear. RESULTS: The reconstruction of the contour of the wing of the nose succeeded in a satisfactory manner. There are no functional restrictions on nose breathing. CONCLUSIONS: The combination of an island advancement flap with a composite graft from the ear for the reconstruction of the alar region is essentially a less invasive operation that can be carried out under local anaesthesia and that represents an addition to the previously stated methods. PMID- 9391568 TI - Upper lip reconstruction with local island flap after neoplasm excision. AB - BACKGROUND: The upper lip is an important esthetic unit of the face and its reconstruction is a big challenge to the dermatosurgeon. OBJECTIVE: The aim of the paper is to present results of upper lip reconstruction for moderately sized defects with single island subcutaneously pedicled flaps, or in combination with additional local flaps. METHODS: Thirteen patients were operated upon mostly for malignant skin neoplasms. Defects of skin only were covered with island flaps planned individually to fit esthetic units of the face. Defects in skin and vermillion were covered with island flaps, and additionally and independently with flaps that reconstructed the vermilion. RESULTS: The healing process was in all the cases but one (partial flap necrosis) free of complications. Follow-up results were also very good functionally and esthetically. PMID- 9391569 TI - Cryosurgery for cutaneous malignancy. An update. AB - BACKGROUND: Cryosurgery is a successful modality for destruction of basal and squamous cell carcinomas. There is a trend toward more aggressive freezing of lesions and treatment of selected difficult tumors. OBJECTIVE: The indications and advantages for cryosurgery, recent treatment techniques, tissue response, complications, and results merit description. METHOD: The standard method of treatment is used with aggressive freezing of the lesion, including a greater lateral spread of freeze and attainment of lower tissue temperatures. RESULTS: The cure rate of cryosurgery is high and the cosmetic results are good to excellent. CONCLUSION: The results of cryosurgery are comparable with other modalities. It may be preferable or advantageous for some patients because of the quickness and safety of the procedure and its low cost. With changes in health care spending due to law or regulation, and within the environment of a managed care system, dermatologists may choose cryosurgery on the basis of outcome and cost. PMID- 9391570 TI - Radiation therapy in skin cancer. A historical perspective and current applications. AB - BACKGROUND: Radiation therapy has been used for skin cancer for nearly a century. During this protracted period, techniques of administering superficial irradiation have developed continuously. More recently, the availability of electron beams for treatment of skin cancer has improved further our ability to treat skin cancer more efficiently and with less toxicity. These include primary skin cancers as well as any metastatic skin lesion. OBJECTIVE: Indications and advantages of modern radiation therapy in skin cancer are confusing at times. In this review, an attempt was made to clarify the role of this discipline in dermatologic use. METHODS: Old literature has been reviewed in order to give an appropriate historical perspective of treatment of skin with irradiation. Literature has also been selected for updating information on current indications and practice of radiation therapy for skin lesions. RESULTS: Radiation therapy is very effective in many situations where other modalities are contraindicated or functionally or cosmetically impairing. With the most efficient methods of fractionation and administration, the control rate of most skin cancers with radiation is as high as 90% or more. CONCLUSION: Modern day radiation therapy is very effective in treatment of skin cancers. Not only the control rate is as good as any other modality, but, with irradiation, cosmetic appearance and function are better preserved under most circumstances. PMID- 9391571 TI - Physician-patient confidentiality. Legal and ethical implications in the setting of basal cell nevus syndrome. AB - Physician-patient confidentiality represents one of the core aspects of any medical practice. Dermatologic surgeons need no advice regarding the privacy of a patient's records. However, occasionally, the boundaries of this confidentiality become obscure. This paper, describing a potential real case scenario, looks at the legal and ethical issues surrounding physician-patient confidentiality. PMID- 9391572 TI - Mohs surgery for nonagenarians. PMID- 9391573 TI - Tattoo formation from suture or from cosmetics? PMID- 9391574 TI - Cochlear implants: what can be achieved? AB - Cochlear implants in children are effective in terms of speech perception and production. Onset of deafness, date of implantation, and duration of deafness are important prognostic factors for long-term results. Sophisticated intraoperative and postoperative device control measurements enable detection of partial or total device failure and calculation of the reliability of the implant. However, the present technological status is far from an optimal device. Future developments will include improvements of both electrode design and speech processing. The number and separation of channels will be increased, and the pattern of stimulation will approach the natural excitation pattern of the auditory nerve. Behind-the-ear implants or totally implantable devices will further increase the acceptance and reliability of this technology. The implant systems will become more versatile and adaptable to the physiological conditions of the individual patient. Apart from these technological improvements, rehabilitation and proper selection of patients will still play a major role in successful implantation in children. Outcome studies are necessary to calculate the cost-benefit ratio and justify a further extension of implantation. We are still on the way toward the artificial ear. PMID- 9391575 TI - The Children Cochlear Implantation Project in Hannover. PMID- 9391576 TI - Basic neurophysiology of cochlear-implants. AB - Cochlear implants work because the fibers of the auditory nerve can be stimulated electrically. Currently used multi-channel electrodes distribute the stimuli to different cochlear places (place principle) and provide information on the fine time structure of the acoustic stimulus thus allowing periodicity analysis by the central nervous system. The paper treats the limitations of current cochlear implant technologies and discusses conceivable improvements. PMID- 9391577 TI - Are spiral ganglion cell numbers important for speech perception with a cochlear implant? AB - OBJECTIVE: Studies of factors affecting the survival of ganglion cells in adults with profound hearing loss and speech perception in cochlear implant users are reviewed in order to assess the hypothesis that ganglion cell numbers have a strong influence on the level of speech perception achieved by adult implant users. DATA SOURCES: All histopathologic data have been published as tables or figures in refereed papers. Speech perception data were collected from published papers and directly from clinics and from Cochlear Corporation's database. STUDY SELECTION: Histopathologic studies with reasonably large groups of patients were selected. The speech perception data were from the largest study available. DATA EXTRACTION: All data used have been published in refereed journals of high repute. No other assessment of quality or validity was available to the author. DATA SYNTHESIS: Trends in the speech perception data were compared qualitatively and quantitatively with trends in the histopathologic ganglion cell data. CONCLUSIONS: The analysis found no strong evidence that spiral ganglion cell numbers have a strong influence on the level of speech perception achieved with a cochlear implant. A possible explanation for this surprising result may be that the minimum number of cells required for good speech perception is quite low, and the majority of implant users exceed this minimum requirement. PMID- 9391578 TI - Intracochlear, electrical, multichannel stimulation effects on the development of auditory system in neonatally deafened kittens. AB - OBJECTIVE: To investigate the effect of chronic electrical stimulation in acoustically deprivated animals during maturation. MATERIALS AND METHODS: Latencies of EABR measurements from acoustically deprivated and acoustically deprivated, but electrically stimulated animals were compared with ABR from normal hearing cats. In addition, morphological analyses of the cochlear nuclei and the auditory cortex and their subdivisions were done. RESULTS AND DISCUSSION: EABR latencies demonstrated that the most peripheral auditory pathway is more independent from the normal auditory or external electrical stimulation than the more central regions. Morphological analysis also demonstrated the reverse of the acoustically deprivation effect during maturation in the auditory cortex via intracochlear electrical stimulation. PMID- 9391579 TI - Ontogeny of electrically evoked brain stem potentials in neonatally deafened gerbils (Meriones unguiculatus) after cochlear implantation. AB - To investigate the ontogeny of the electrically evoked auditory brainstem response (E-ABR) we used an animal model of neonatally deafened gerbils with an intracochlear implanted electrode. The E-ABR were recorded in three groups: normal hearing animals (NORM) and binaural deafened without chronical electrostimulation (BD) in comparison to binaural deafened animals with chronical stimulation (BDS). In group BD the deafening lead to inter-peak-latency II-V increase and histological degeneration in the Cochlear Nucleus. In chronical stimulated animals (BDS) we could not observe significant differences compared to unstimulated gerbils until the day after birth. In further studies longer stimulation periods, recordings of electrically evoked middle latency response as well as histological examinations of more central parts of the auditory pathway will be included. PMID- 9391580 TI - Optical imaging of cat auditory cortical organization after electrical stimulation of a multichannel cochlear implant: differential effects of acute and chronic stimulation. AB - OBJECTIVE: To study the effects of electrical stimulation on cortical activation patterns. MATERIALS AND METHODS: Optical imaging of auditory cortex in cats that are acutely and chronically electrically stimulated with cochlear implants. RESULTS: Chronic electrical stimulation results in expansion of cortical territory and overlap. CONCLUSION: Effects of chronic electrical stimulation are comparable to use-dependent cortical plastic reorganization. PMID- 9391581 TI - Acute effects of electrical intracochlear multichannel high-rate stimulation of the auditory nerve of cats. AB - OBJECTIVE: To investigate the short-term effects of electrical stimulation of the cochlea in cats. MATERIALS AND METHODS: Deafened cats were implanted with human cochlear electrodes, electrical stimulation at clinical levels was given, and evoked auditory brain stem responses (EABR) were recorded. RESULTS AND DISCUSSION: No long-term reduction in the excitability of the auditory nerve or in brain stem responses was seen. PMID- 9391582 TI - Meriones unguiculatus (Gerbil) as an animal model for the ontogenetic cochlear implant research. AB - The rehabilitation of prelingually deafened children using cochlear implants still raises several basic problems, e.g. growth of the skull and maturation of the auditory system. Systematic research in these problems requires a reliable animal model. The gerbil (Meriones unguiculatus) was investigated in respect of its suitability as an animal model for ontogenetic cochlear implant research. Gerbil pups were deafened by a single intracochlear application of neomycine on their 14th day after birth before the onset of natural hearing and implanted with an animal specific cochlea implant at day 30 after birth. To maintain the biological relevance the social sounds of gerbils were frequency transformed and used in daily electrostimulation until the 90th day after birth. As well established animal in hearing research the gerbil appears to be an appropriate animal model for ontogenetic cochlear implant research. PMID- 9391583 TI - Development of the evoked auditory brain stem response amplitude peak IV during chronic electrical, intracochlear, multichannel high-rate stimulation. AB - OBJECTIVE: To investigate the effect of high stimulation rates on the developing auditory system. MATERIALS AND METHODS: Neonatally deafened kittens with human intracochlear electrode implants received sustained electric stimulation. RESULTS AND DISCUSSION: No loss of brain stem responses was observed. PMID- 9391584 TI - Histologic and immunohistochemical investigations on the development of the auditory brain stem modified by auditory deprivation. PMID- 9391585 TI - Measurements of threshold shifts observed when using normal and preformed electrodes. AB - OBJECTIVE: To investigate the efficacy of short-term electrical stimulation with different human electrodes using high stimulation rates and different electrode designs. MATERIALS AND METHODS: Human electrodes of two different designs were implanted in cats, and electrically evoked auditory brain stem responses (EABR) were recorded. RESULTS AND DISCUSSION: Thresholds from the EABR recordings at the apical end of the electrode array were comparable in both types of implants. There were significantly higher thresholds at the basal end than at the apical end of the electrodes in both types of electrodes. PMID- 9391586 TI - Effect of high-frequency electrical stimulation of the auditory nerve in an animal model of cochlear implants. AB - HYPOTHESIS: Electrical stimulation of the cochlea at high rates induces significant adaptation of the auditory nerve. BACKGROUND: A new development of cochlear implants is the use of speech processors delivering electrical pulses on the implanted electrodes at high rates, such as 1,000 pulses per second (pps) and above. Such a stimulation mode allows subjects with cochlear implants to reach excellent understanding of speech. METHODS: Long Evans-rats received implantation of stimulating electrodes in the left cochlea. Two hundred-millisecond trains of short (20 microns) monophasic pulses were delivered in 50% duty cycle at 500 microA above threshold. The pulse rate in the train was increased from 100 pps to 1,500 pps. Electrically evoked auditory brainstem responses (EABR) were recorded. The amplitude of the compound action potential of the auditory nerve to each single pulse in the train was measured as the first vertex positive wave (WAVE I) of the EABR. RESULTS: At 100 and 200 pps, WAVE I amplitudes to each pulse were large and remained stable throughout the pulse train. For increasing pulse rates, WAVE I amplitudes progressively decreased during the first 40 to 50 ms of the train and reached 80% at 300 pps to 15% at 1,500 pps of the maximal amplitude observed for the first pulse in the train. CONCLUSIONS: The decrease of the WAVE I amplitude in response to high-rate pulsatile stimulation reflects an adaptation of the auditory nerve due, at least in part, to the refractory period of auditory nerve fibers. PMID- 9391587 TI - Temporal representations with cochlear implants. AB - OBJECTIVE: To record and characterize intracochlear evoked potentials (EPs) for a variety of electrical stimuli in studies with cochlear implant patients. METHODS: Recordings were made with patients having direct percutaneous access to their implanted electrodes. Intracochlear voltages were recorded via unstimulated electrodes. The stimuli included trains of identical pulses, with pulse rates ranging from 100 to 4065/s, and a modulated pulse train produced by a single channel speech processor, with the pulse rate of 824/s. RESULTS: Magnitudes of EPs for each pulse in trains of identical pulses were uniform for pulse rates below about 200/s. For rates between about 400 and 1000/s, an alternating pattern of EP magnitudes was observed, with relatively large EPs following the odd numbered pulses. For rates between about 1000 and 3000/s, more complex patterns were observed. After the first millisecond of each train at even higher rates, uniform EPs again were observed across pulses, although the absolute magnitude of the EPs was much lower than that observed for low rates of stimulation. The approximate rates corresponding to boundaries between these different regions varied among subjects and among electrodes within subjects. EP magnitudes for the modulated pulse train reflected the gross periodicity of the modulation waveform but did not reflect temporal details within the periods. CONCLUSIONS: Population responses of the human auditory nerve, as indicated by EP magnitudes, reflect the amplitudes of electrical pulses for pulse rates below about 200/s and above about 3000/s. Use of intermediate rates may introduce distortions in the transmission of stimulus information with cochlear implants. PMID- 9391588 TI - Results from a pilot study using the nucleus CI24M/SP5 cochlear implant system. AB - OBJECTIVE: To conduct a pilot study in adults with the Nucleus CI24M/SP5 cochlear implant system. PATIENTS AND METHODS: Eight postlingually deafened adults who had received little or no benefit from conventional hearing aids, equipped with the Nucleus CI24M/SP5 cochlear implant system. RESULTS AND CONCLUSIONS: The results indicate that most of the subjects were able to perform well in speech recognition tests. The test performances appeared to be strongly affected by the duration of deafness. The speech processor's four user-selectable program memories have been extremely useful for the subjects to evaluate variations to the speech coding strategies in ordinary surroundings outside of the laboratory. The telemetry functions of the new implant provide a set of useful clinical and research tools for gathering greater insights into the in-situ operation of the implant. PMID- 9391589 TI - The advanced Combi 40+ cochlear implant. AB - A 12-channel cochlear implant (CI) for high-rate pulsatile stimulation strategies is presented. Symmetric biphasic current pulses can be generated up to a maximum pulse repetition rate of 18.18 kpulses/second. The stimulation pulse amplitude can be selected within 1.5 microA-1.5 mA. Data and power are transcutaneously transferred using a single radiofrequency (RF) channel. A fully digital data transfer format is employed at an overall data rate of 600 kBit/second. The implant contains a single mixed analog/digital CMOS-ASIC (Application Specific Integrated Description) for data synchronization and stimulus generation. Stimulation signals are applied via a monopolar intracochlear multichannel electrode. Output capacitors for each channel are employed for safety reasons. A self-calibrating back telemetry system is included for estimating the channel impedances and field distribution along the electrode array. Dimensions of the ceramic package of the implant are only 33.50 x 23.40 x 3.95 mm3. PMID- 9391591 TI - Electrodes for ossified cochleas. PMID- 9391590 TI - Nucleus double electrode array: a new approach for ossified cochleae. AB - INTRODUCTION: The ossified cochlea is still a special surgical issue that requires a special surgical procedure. The current cochlear implants only have one electrode lead, which can be placed only partially in the drilled out basal turn. The small number of used active electrodes leads to worse performance as compared with patients with full insertion. METHODS: To overcome this limitation, a special electrode was developed consisting of two arrays. One array with 11 active electrode rings is placed in the drilled out basal turn, the second array with 10 active electrodes in the opened second turn. The number of inserted electrodes can be significantly increased. The surgery is similar to that in nonossified cochleae. After the posterior tympanotomy, the bridge is removed and the incus is located. A cochleostomy is performed at the basal turn and the new built tissue removed. A second cochleostomy is placed below the cochleariform process. In most cases, the second turn is not obliterated and the second electrode array can be fully inserted. RESULTS: The surgical procedure was in all nine cases uneventful. Intraoperative stapedius reflex could be recorded with elevated thresholds. The wide variety of stimulation modes and sites allows an individual fitting to maximize the performance. All patients show a gap in the pitch scale between the apical and the basal array. The pitch variation is much smaller in the apical array. All patients have some benefit from the additional apical array and an improved performance. CONCLUSION: The nucleus double electrode array is an advanced treatment option for patients with ossified cochleae. The receiver/stimulator is a regular nucleus cochlear implant. PMID- 9391592 TI - Pediatric experience of the reliability of the nucleus mini 22-channel cochlear implant. PMID- 9391593 TI - Evaluation of noise reduction systems for cochlear implant users in different acoustic environment. AB - Evaluation of two different noise reduction algorithms for speech intelligibility enhancement in cochlear implant (CI) users is described in this report. The algorithms accomplish sophisticated interchannel processing of the noisy speech signals, picked up with two microphones, to form an improved monaural output signal, which is directly fed into the auxiliary input of the CI speech processor. Speech intelligibility tests were carried out in different realistic everyday life listening conditions to provide general and expressive performance assessment. Extensive tests in four CI users showed considerable speech intelligibility improvement using these noise reduction systems in adverse everyday life listening conditions. PMID- 9391594 TI - Magnetless cochlear implant: relevance of adult experience for children. AB - A method of implanting magnetless cochlear implants without alignment problems is outlined and the first adult experience is presented. Some special considerations relevant for implanting such an implant in children are discussed. PMID- 9391595 TI - Evaluating the plasticity potential of the auditory brain stem nucleus in the rat. AB - OBJECTIVE: To study the adaptation of the auditory brainstem to auditory loss. STUDY DESIGN: Growth-associated protein 43 (GAP-43) immunoreactivity was studied in in rats whose cochleas had been removed. RESULTS AND DISCUSSION: Neurons in the lateral superior olive were found to synthesize GAP-43 in a pattern that paralleled the changes in GAP-43 immunoreactivity in the cochlear nucleus after cochlear ablation. These findings suggest that new patterns of synaptic communication can be established after damage to the cochlea. PMID- 9391596 TI - Submillimeter imaging and reconstruction of the inner ear. AB - OBJECTIVE: To present new radiological developments using high-resolution magnetic resonance imaging (MRI). MATERIALS AND METHODS: Using heavily T2 weighted sequences at a 1.5 Tesla scanner, maximum-intensity projections (MIP) of the inner ear were generated. The imaging time was less than 20 minutes, and imaging could be performed with adults and children of all age groups alike. This method enables us to visualize and identify the different neural structures of the internal auditory canal. Aplasia or schwannoma of a single or a variation of nerves could be clearly demonstrated. RESULTS: Three-dimensional reconstruction enabled unprecedented clear and precise presentation of the fluid content of the inner ear. The size and shape of 2 to 2 1/2 turns of the cochlea could be routinely demonstrated and analyzed in 45 subjects. Eight patients subsequently received an intracochlear implant electrode, and the intraoperative findings correlated with the imaging. CONCLUSION: The most recent high-resolution MRI techniques provide reliable visualization of submillimeter anatomical structures of the inner ear and auditory nerve. PMID- 9391597 TI - Large vestibular aqueduct syndrome and its implication for cochlear implant surgery. AB - The large vestibular aqueduct syndrome (LVAS) is commonly found when presurgical computed tomography is performed in advance of cochlear implantation. We present data on two patients with LVAS, in whom computed tomography (CT) and magnetic resonance imaging (MRI) was performed. Cochlear implantation when these malformations are present may be complicated by an intraoperative gusher, but it appears to be safe and produces a favorable outcome. PMID- 9391598 TI - Intact canal wall drill-out procedure for implantation of the totally ossified cochlea. AB - OBJECTIVE: To describe a simplified drill-out technique for insertion of a multichannel electrode in the completely ossified cochlea without radical mastoidectomy and obliteration. STUDY DESIGN: Description of a new surgical technique and case report. SETTING: Temporal bone dissection laboratory and tertiary referral center. PATIENTS: Adult and pediatric cochlear implant (CI) recipients. MAIN OUTCOME MEASURES: Access for circum-modular drill-out and electrode insertion without radical mastoidectomy and adequate function of multichannel CI. RESULTS: Dissection of 10 cadaver temporal bones demonstrated feasibility of this technique. Highlights include facial recess cochleostomy and 8 mm tunnel; elevation of superiorly based tympanomeatal flap; removal of incus, cochleariform process, and tensor tympani; and identification of carotid canal and use of facial nerve monitor. A case report of an 11-year-year old child with total cochlear ossification and previous failure of a short (8 electrode) CI electrode insertion is presented. Complete insertion of a 22-channel electrode was successful and open-set word recognition is commencing. CONCLUSIONS: The canal wall-up drill-out procedure allows complete electrode insertion without mastoid obliteration in patients with obliterated cochleas. Appropriate attention to the carotid artery and facial nerve is essential. PMID- 9391599 TI - Cochlear implant reimplantation. AB - The objective of this study was to determine whether insertion length and number of active channels remained the same after reimplantation of a cochlear implant. A retrospective case review of 170 consecutively implanted multichannedl cochlear implants was conducted. Seventeen of these devices had to be replaced. Data were analyzed for the Nucleus cochlear implant users who were reimplanted in the same ear. For most subjects, insertion length and number of channels remained unchanged, but a few subjects experienced substantial decreases. When the whole group was considered, a small but statistically significant drop was noted for both parameters. In conclusion, although reimplantation is technically possible, the first procedure provides the optimal surgical environment. PMID- 9391600 TI - Complications of cochlear implant surgery in children. AB - OBJECTIVE: To evaluate the intraoperative problems and postoperative complications in children receiving cochlear implants. STUDY DESIGN: A retrospective analysis of the clinical records of 366 children, aged 1 to 14 years, who had received cochlear implants. RESULTS: Intraoperatively, obliteration of the cochlea occurred in 66 patients, cochlear dysplasia in 8, and CSF leakage in 7, and nearly 5% of patients had signs of infection in the mucosa of the middle ear. Postoperatively, early complications occurred in 1% to 2.5% of patients: flap complications, electrode dislocation, facial nerve problems, and incorrect insertion of the electrode. Delayed complications included otitis media and stimulation of the facial nerve. CONCLUSION: Proper preparation of the implantation site, experienced and well-trained surgeons, and awareness of the operative and postoperative risks are necessary. PMID- 9391601 TI - Malformations in cochlear implant patients. AB - OBJECTIVE: To report on cochlear implantation in children with bony inner ear malformations. PATIENTS: 30 children with bony inner ear malformations who have received cochlear implants. INTERVENTIONS: High-resolution spiral computed tomography is used to identify malformations. Magnetic resonance imaging is used to detect the presence of an acoustic nerve and determine the integrity of the auditory pathway and central nervous system structures. Both imaging techniques may be used intraoperatively, as well as facial nerve monitoring and electrical auditory brainstem response monitoring. Three-dimensional reconstructions are helpful in preoperative planning. Large vestibular aqueducts and vestibular malformations can be successfully managed. RESULTS: Postoperative results have been encouraging, although children with malformations tend to occupy the lower third of rehabilitation results of all children with implants. PMID- 9391602 TI - Surgical techniques for cochlear implantation of the malformed inner ear. AB - OBJECTIVE: The objective was to describe surgical techniques helpful in implanting children with inner ear malformations. STUDY DESIGN: This was a retrospective chart review and description of surgical techniques in the setting of a tertiary referral center. PATIENTS: The study population was composed of 10 children with inner ear deformities who received 22-channel implants. RESULTS: The primary surgical challenges encountered in these procedures include complete electrode insertion, cerebrospinal fluid gusher, identification of cochleostomy site in the absence of the round window and aberrant facial nerve, and fixation and stabilization of the electrode. CONCLUSIONS: The techniques described allow safe and effective insertion of multichannel electrodes in patients with inner ear malformations. PMID- 9391603 TI - Medical, surgical, and technical complications with the COMBI-40. AB - We present the first report on complications of cochlear implantation with the COMBI-40 (Med-el, Innsbruck, Austria). Between January 1995 and May 1996 325 devices had been implanted by 58 different surgeons. Complications were reported with the help of standardized complication report form. The overall rate of complications was 4.6%. Most common problems were flap necrosis and incorrect positioning of the electrode. No technical failures occurred. The incidence of complications was lower than those reported by other authors. PMID- 9391604 TI - Surgical aspects of cochlear implantation in young children: a review of 115 cases. AB - A review of the surgical complications of the first 115 patients who received a cochlear implant in this program demonstrate a negligible complication rate in the perioperative period and a low complication rate in the long term. The surgical technique is described in detail and the reasons for this low complication rate are analysed. PMID- 9391605 TI - Selection criteria for cochlear implants in children. AB - OBJECTIVE: To determine the selection criteria for cochlear implantation in children. SETTING: Hospital pediatric implant center. PATIENTS AND INTERVENTIONS: Selection of patients depends on medical evaluation, audiometric data, speech discrimination, communication skills, cognitive skills, and psychosocial factors. Patient selection is based on tonal audiometry, computed tomography, magnetic resonance imaging, and electrophysiologic tests. Side of implantation is chosen according to cochlear structure, duration of deafness, and dominant handedness. RESULTS: Ninety-eight cochlear implantations with the Nucleus multichannel implant have been performed at this center since 1990. CONCLUSION: The selection of children for cochlear implantation require close collaboration between the pediatric surgical team, the educational team, and the family. PMID- 9391606 TI - Preoperative radiologic evaluation in cochlear implantation. AB - AIM OF STUDY: Assess the value of computed tomography (CT) in the evaluation of abnormalities in the cochlea and auditory pathways. MATERIAL AND METHODS: We used CT to evaluate 108 children before cochlear implantation surgery. Children's ages at implantation ranged from 21 months to 16 years (mean age, 5.4 years). The etiology of deafness was meningitis in 44 children (40.8%), congenital in 51 (47.2%), and other in 13 children (12%). RESULTS: Eighteen of the 108 (16.6%) children and 34% of the postmeningitic children were found to have at least partial obliteration of the cochlea. Two (2%) children had congenital malformations of the cochlea and 12 children (11.1%) had abnormalities in the brain CT-scan. CT diagnostic values in postmeningitic children regarding cochlear obliteration were accuracy, 75%; sensitivity, 62%; specificity, 82%; positive predictive value, 66.6%; and negative predictive value, 79.3%. In six (20.6%) of postmeningitic children with normal CT-scans, some scala tympani drillout was required. CONCLUSION: CT-scan is capable neither of predicting with certainty the presence of minor degrees of cochlear obliteration nor of specifically imaging either the auditory nerve or its central connections. PMID- 9391607 TI - A European perspective on pediatric cochlear implantation, rehabilitation services, and their educational implications. AB - OBJECTIVE: To assess the educational implications of pediatric cochlear implantation from the perspective of the implant team. METHODS: Coordinators of pediatric cochlear implant teams throughout Europe took part in a survey using forced-choice questions. Fifty-four centers were originally sent the questionnaire; 41 centers replied. RESULTS: Of 504 children planned to receive cochlear implants in Europe during 1996, 54% (273/504) were aged 2-5 years and 12% (60/504) aged 0-2 years, indicating a trend toward pediatric implantation in younger children. There is a strong commitment to rehabilitation in the teams; 66% (27/41) employ a teacher of the deaf, the ratio of medical/audiological to rehabilitation personnel is 1:2, and 76% (31/41) of the implant teams visit local educators. Of all the children receiving implants to the date of this report, 23% were considered to be in unfavorable educational environments; these were environments where children were taught with an emphasis on sign language and little expectation from audition, and mainstream provision without support from experienced teachers of the deaf. CONCLUSION: There is a high staff input to children with cochlear implants from implant rehabilitation personnel over and above the input received in the educational environment. Hence, it is important for the school and the implant team to mutually agree on their shared responsibilities. Moreover, as the provision of service is variable and inconsistent, the development of guidelines for practice in each country should ensure consistency of rehabilitative and educational support to children with cochlear implants. PMID- 9391608 TI - Use of a parent-report scale to assess benefit in children given the Clarion cochlear implant. AB - OBJECTIVE: To assess the auditory skills in everyday situations of prelingually deafened children with Clarion cochlear implants compared with hearing aids used preoperatively. STUDY DESIGN: The Meaningful Auditory Integration Scale (MAIS) was used to determine the preoperative and postoperative auditory skills of the children. RESULTS AND DISCUSSION: After implantation, the children showed improvement in three skill areas: bonding to the device, spontaneous alerting to sound in everyday situations, and ability to derive meaning from sound in the environment. PMID- 9391609 TI - Assessment of auditory skills in hearing-impaired children: theoretical foundations and a practical approach. AB - A proposal of a performance profile for the assessment of auditory skills of hearing-impaired children with cochlear implants or hearing-aids is described here. The associated tests have been realized either with conventional material or on the Auditory Visual Test and Therapy System developed by the Aachen research group. Acoustic stimuli and answering tasks have been chosen according to the special needs of hearing-impaired children in the age range of 3-6 years of developmental age. Two tests will be described exemplary. PMID- 9391611 TI - Keynote lecture: objective measures. AB - The number of young children receiving cochlear implants for management of profound hearing loss is increasing world-wide. In these children, objective measures play a vital role both before and after implantation. They have the capability to assess the degree of hearing loss, guide the selection of the ear and patient for implantation, and provide valuable assistance with the tuning of the device in the difficult-to-test and very young children. There is considerable opportunity for further development of these techniques in the future and, in particular, for accessing central perception and cognition. PMID- 9391610 TI - The HSM sentence test as a tool for evaluating the speech understanding in noise of cochlear implant users. AB - The German HSM Sentence Test, available on compact disc (CD), consists of 30 lists of 20 everyday sentences. It was developed in the desire to have a sufficient number of test sentences for the repeated evaluation of speech understanding of CI users. A noise with speech-shaped spectrum on the CD allows the evaluation of speech understanding in noise. With the HSM Sentence Test we evaluated the speech understanding of participants of the Combi-40 European Multicentric Study. Results are shown for sentences presented without noise and with signal-to-noise ratios of 15, 10, 5 and 0 dB. PMID- 9391612 TI - Comparison of preoperative electrostimulation data using an ear-canal electrode and a promontory needle electrode. AB - OBJECTIVE: To compare the electrical stimulation results of the ear-canal electrode with those of a promontory needle. PATIENTS AND METHODS: In thirty three adult patients, the ear-canal electrode test was compared with the needle electrode promontory test with respect to sound perception, rhythm detection, frequency, and disturbing side effects. RESULTS: The ear-canal electrode test, in comparison with the needle electrode promontory test, resulted in vibrotactile sensation in addition to auditory sensation in some patients, less auditory fatigue, higher threshold levels, and lower discomfort levels. CONCLUSION: Reliable assessment of deafness in children requires, in addition to electrical stimulation, determination of the electrical evoked auditory brainstem response with the patient under anesthesia. PMID- 9391613 TI - Electrically evoked auditory potentials: current clinical applications in children with cochlear implants. AB - OBJECTIVE: To summarize the current applications of auditory evoked potential in children with cochlear implants and candidates for implantation. PATIENTS AND METHODS: Perioperative transtympanic EABR is used routinely for ear selection and to establish the electrical stimulability of the ear intended to be implanted. The perioperative transtympanic EABR is supplemented with EABR obtained immediately following the insertion of the electrode array and the seating of the implant's receiver. Postoperatively, EABR and averaged electrode voltages are used effectively to properly adjust the implant stimulus parameters and to determine whether the implant is functioning adequately. Postoperatively, cognitive evoked potentials to speech and tonal stimuli may also be obtained. RESULTS: EABR results have contributed to effective implant placement and function. There were several significant correlations between speech recognition and cognitive evoked potential. CONCLUSION: These measures help assure proper implant function and effective stimulus delivery. PMID- 9391614 TI - Intraoperative test of auditory nerve function. AB - OBJECTIVE: To develop a preoperative objective test of auditory nerve function for the assessment of cochlear implant candidacy, especially in children. PATIENTS AND METHODS: We stimulated electrically with a ball electrode before insertion of the implant. First the stimulus was applied bipolar between the promontory and the round window. To record auditory brain stem responses (EABRs) very high stimulus intensities were needed, which was not possible in all patients. RESULTS: By stimulation in the basal turn of the cochlea, evoked potentials could be derived. Although in some patients the facial nerve was stimulated as a side effect, auditory evoked potentials could be recorded. A facial muscle artifact can be differentiated from the EABRs by latency and the slope of the input-output function. CONCLUSION: For the present, the only reliable test seems to be the EABR recording stimulated within the cochlea. PMID- 9391615 TI - Monitoring the electrode position during acoustic neuroma surgery. AB - OBJECTIVE: To obtain information about the auditory brain stem responses during auditory brain stem implantation. SETTING: Operating room during acoustic neuroma surgery. METHODS: Electrical stimulation of the auditory system during acoustic neuroma surgery, by placement of a monopolar or bipolar electrode on the nerve or nerve entry zone of the brain stem, and monitoring of the evoked auditory brain stem responses (EABR) recorded from the scalp. In some patients, a multichannel silicon electrode array was placed at the foramen of Luschka. Biphasic rectangular current pulses were applied, and EABRs were recorded. RESULTS: Usually the derived potentials consisted of three peaks with a latency below 4 ms. Sometimes we got a complex of two or more peaks. The interpeak interval between the first and second peak was about 0.7 to 1.0 ms, independently of the stimulating electrode position, but the absolute latency of the first peak increased from a minimum of 0.7 ms stimulated at the foramen of Luschka to a maximum of 1.3 ms stimulated at the nerve. PMID- 9391616 TI - Assessment of electrostimulation in children supplied with cochlear implants. AB - OBJECTIVE: In children supplied with cochlear implants (CIs), there is an urgent need for objective tests for the assessment of electrostimulation. SETTING: An optimized procedure for intraoperative and post-operative applications with short duration test time, based on the measurement of the electrical farfield picked up from surface electrodes on the scalp, is presented; therefore, assessment of stimulation is independent of the stimulating electronic circuitry. OUTCOME: The setup was designed for the COMBI40 multichannel CI using a synchronized sampling technique of the amplified signals. The evaluation procedure includes examination of the time course of the signal and calculation of the input output function. With these options, a straightforward judgment of the electrical performance of the implant in situ, reflecting the input to the auditory nerve, is provided. PMID- 9391617 TI - Electrode and device problems: manifestation and management. AB - A total of 119 children were implanted with the Nucleus 22 implant on the Nottingham program by March 1996. Twenty-five (i.e. 21%) of these had an electrophysiologically confirmed fault on at least 1 channel and 6 (i.e. 5%) had experienced total device failure. How these problems were first manifested and what the subsequent effects were on the child and family were determined by means of questionnaires and detailed examination of clinical notes. Our findings indicated that 76% of all internal device faults were initially detected in tuning or electrophysiological measurement. All parents expressed deep anxiety about the threat of device failure; cases of total failure occurring over a long period of time resulted in the greatest distress and effect on the family. Recommendations are made for companies to assist clinics in minimizing such effects. PMID- 9391618 TI - Which sensitivity setting should a child use? AB - The influence of the sensitivity setting of the cochlear implant speech processor on speech recognition is investigated with 15 experienced postlinguistically deafened adult cochlear implant users. We describe the use of loudness scaling in the course of speech processor fine tuning and checkup of the speech processor map to demonstrate how their speech processors are programmed. The Freiburg monosyllabic word test, a standardized German open set speech intelligibility test, is used in silence and the very difficult Gottingen sentence test is used in silence and noise to measure speech recognition scores as a function of sensitivity setting. The median of the optimal sensitivity setting was four steps higher than the value recommended by the manufacturer. In noisy everyday hearing situations patients generally use a sensitivity setting one step lower than the one we found to be optimal for openset word understanding under test room conditions. With children who cannot select optimal speech processor settings by themselves, we do not recommend the use of noise reduction and set the sensitivity three steps higher than the one recommended by the manufacturer. PMID- 9391619 TI - Optimization of channel number and stimulation rate for the fast continuous interleaved sampling strategy in the COMBI 40+. AB - OBJECTIVE: To investigate the interrelation between number of channels and stimulation rate in the continuous interleaved sampling strategy (CIS). SUBJECTS AND METHODS: Three of the first recipients of the new COMBI 40+ cochlear implant participated in consonant, vowel, number, and sentence tests. Speech understanding was evaluated for different combinations of number of active channels from two to twelve and stimulation rate per channel between 1,515 and 9,090 pulses per second. RESULTS: The results indicate that the optimum number of active channels is not necessarily the maximum number of usable channels. PMID- 9391620 TI - Evaluation of electrically elicited stapedius reflex threshold measured through three different cochlear implant systems. AB - OBJECTIVE: To evaluate intraoperative electrically elicited stapedius reflex thresholds (ESRTs) measured through three different cochlear implant systems: the Nucleus Mini 22, the Clarion Enhanced Bipolar, and the Med-El Combi-40. SUBJECTS AND METHODS: Relations between intraoperative ESRT and postoperative maximum comfort level (MCL) were examined in seven children (4 Nucleus, 2 Clarion, and 1 Med-El) and one adult (Clarion). RESULTS: Preliminary results indicated most ESRTs were either higher or both higher and lower (across the electrode array within a subject) than MCLs. All systems provided satisfactory means for measuring ERSTs. CONCLUSION: It is recommended that hand-held systems have a direct readout to the programming station and that audio and visual feedback be improved for all units. PMID- 9391621 TI - Long-term stability of fitting parameters with the COMBI 40. AB - Having experience with the COMBI 40 for over two years, it is now possible to look at the development of patient fittings over a longer period of time. In this study we look at patient fitting data (thresholds, dynamic ranges, etc.) at two days, one month, three months, six months and twelve months post first fitting. Subjects are CI-users participating in the COMBI 40 European Multicentric Study. Long-term fitting parameters show a favourable development. Stable fitting parameters and the safety features implemented in the COMBI 40 serve as a basis for a reliable long-term speech understanding. PMID- 9391622 TI - Amplitude modulation following response in children as a clinical audiometric tool. AB - The current study was designed to investigate the clinical application of amplitude modulation following response (AMFR) in cochlear implant candidates. A new digital signal processor (DSP)-assisted PC-based hardware and software was developed to perform both simultaneous generation of amplitude-modulated stimuli and the recording, and synchronized signal processing of the electrode signals. Our first results show that AMFR can be recorded in adults as well as in children without any contamination by response-like stimulus artifacts. Very high sound pressure levels can be applied, allowing frequency-specific assessment of residual hearing. Response threshold detection, using spectral analysis, proved to be superior compared to visual evaluation of average time waveforms. PMID- 9391624 TI - Single electrode maps in device troubleshooting. AB - This article presents case studies in which single-channel mapping was used with patients who were experiencing discomfort while using their cochlear implant devices. Repeated psychophysical testing together with integrity testing had failed to locate the source of the problem in each of the described cases. Single channel mapping was then used as another means of device troubleshooting. Single channel maps were created for each electrode across the array. In each case, the patient was able to identify the offending electrode(s) during the presentation of speech stimuli, whereas the problem had not been evident during psychophysical testing with pulse stimuli. Eliminating these electrodes from the map alleviated the problems experienced by these implant users in each case. PMID- 9391623 TI - Speech intelligibility as a function of the number of channels of stimulation for normal-hearing listeners and patients with cochlear implants. AB - OBJECTIVE: One goal was to determine for normal-hearing listeners the number of channels of stimulation necessary to achieve a high level of speech understanding. The second goal was to determine whether patients with a six channel cochlear implant could achieve the same level of speech understanding as normal-hearing subjects listening to speech processed through six channels. METHODS: Speech signals were processed, for normal-hearing listeners, either in the manner of cochlear-implant processors with 2-9 fixed channels, or in the manner of a processor which picked, on each update cycle, 6 of 16 channels. RESULTS: For the most difficult test material eight fixed channels were necessary to achieve the level of performance achieved with the "n of m" processor. Some cochlear implant patients with a six-channel continuous interleaved sampling processor achieved the same level of performance as normal-hearing subjects listening to speech via six channels. CONCLUSIONS: A signal processor for cochlear implants with eight channels should produce the same level of intelligibility as a processor with many more channels. Processors using continuous interleaved sampling technology can provide a signal which results in the same level of speech understanding as normal, acoustic stimulation. PMID- 9391625 TI - Auditory event-related potentials in post- and prelingually deaf cochlear implant recipients. AB - The development of central auditory functions in cochlear implant (CI) patients was studied over six months of rehabilitation. Examinations were performed beginning with the first week after processor calibration, and in monthly follow up sessions thereafter. The subjects were given a simple auditory perception task (detection of a 400 Hz and a 1450 Hz tone), as well as an oddball-paradigm (detection of one of the tones as a rare deviant). Auditory evoked potentials, reaction time and errors were recorded. Results from five patients, two postlingually deaf and three prelingually deaf CI recipients are shown. Generally, in the auditory evoked potentials of patients a shortening of N100 latency towards those of subjects with normal hearing was seen from month to month. However, in the prelingually deaf patients this effect was weaker and more variable over time. Three CI recipients showed a P300 component in the oddball paradigm in correlation with their performance. Two prelingually deaf patients failed to show a P300 in the oddball-paradigm. For both components, the N100 and the P300 we found a larger spreading over the skull in the patients compared to a normal hearing person. The results show that from the very first days after initial processor fitting prelingually and postlingually deaf CI recipients may show cortical correlates of stimulus processing and discrimination. For some components of the auditory evoked potentials an initial temporal change but a maintained larger spreading over the skull was seen. PMID- 9391626 TI - Intraoperative electrical stimulation of the stapedius reflex in children. AB - OBJECTIVE: To describe the use of intraoperative electrical stimulation on the stapedius reflex in children. SETTING: Hospital cochlear implantation center. PATIENTS AND METHODS: Eighteen children under the age of 10 years, who have undergone cochlear implantation, have been examined during surgery with the electrically elicited stapedius reflex in the monopolar stimulation mode. RESULTS: A stapedius reflex could visually be detected for all subjects. CONCLUSION: The responses serve to test the implant, provide guidance in avoiding overstimulation of the child when the device is switched on, and estimate the optimal C-levels. PMID- 9391627 TI - Electric auditory brain stem response in pediatric patients with cochlear implants. AB - OBJECTIVE: To introduce four comprehensive electrical auditory brain stem response (EABR) parameters that objectively measure the input-output function and may be the base of comparison in related studies. MATERIALS AND METHODS: In 53 children (106 ears), recordings of the EABR evoked by electrical stimulation at the promontory were made at the time of surgery after the child was anesthetized and before cochlear implantation. RESULTS: Of the 106 ears studied, 81 (76.4%) produced clearly defined responses. These responses were used to develop a package of four comprehensive EABR parameters (slope, maximal slope, relative growth rate, and maximal relative growth rate) that measure objectively the input output function. The methods of calculation are described in detail. CONCLUSION: These parameters may help us to refine and make more consistent the subjective EABR evaluation. They will also enable a comparison of the results from different cochlear implant centers and promote the progress of related research. PMID- 9391628 TI - Phoneme acquisition in the first 4 years of implant use. AB - OBJECTIVE: To investigate the phonetic inventory development in a group of profoundly hearing impaired children fitted with the 22-electrode cochlear implant (Cochlear Ltd, NSW, Australia) at < or = 5 years of age. BACKGROUND: The cochlear implant provided access to auditory perceptual information that was not previously available. Investigation into the speech production skills of these children postimplant is of interest because the speech of these young profoundly hearing-impaired children is in a constant state of development. METHOD: Phonetic inventories of nine children were monitored at preimplant and over the first 4 years of implant use using spontaneous speech samples collected at regular intervals for each child. Progress of phoneme acquisition was measured using two different criteria: targetless and target production. RESULTS: At 4 years postimplant, 87% of all phonemes had been acquired as targetless productions and 62% of all phonemes had been acquired as target productions. All monophthongs were acquired as target phonemes, but only 38% of the diphthongs and 54% of all consonants were acquired as targets over the time of the study. The average time taken for a phoneme to progress from targetless acquisition to target acquisition was 21.6 months, although variation among phonemes was evident. CONCLUSIONS: Overall, the data suggest similar trends in the order of phoneme acquisition when compared to normally-hearing children, although it would appear that the process of acquisition in children with cochlear implants occurs at a slower rate. PMID- 9391629 TI - Speech perception results for children with implants with different levels of preoperative residual hearing. AB - OBJECTIVE: Many reports have established that hearing-impaired children using the Nucleus 22-channel cochlear implant may show both significant benefits to lipreading and significant scores on open-set words and sentences using electrical stimulation only. These findings have raised questions about whether severely or severely-to-profoundly deaf children should be candidates for cochlear implants. To study this question, postoperative results for implanted children with different levels of preoperative residual hearing were evaluated in terms of speech perception benefits. STUDY DESIGN/SETTING: A retrospective study of the first 117 children, sequentially, to undergo implantation in the Melbourne and Sydney Cochlear Implant Clinics was undertaken. All children had been assessed by and received their implants in a tertiary referral centre. MAIN OUTCOME MEASURES: To assess aided residual hearing, the children were grouped into four categories of hearing on the basis of their aided residual hearing thresholds measured preoperatively. To assess benefits, the scores of children on standard speech perception tests were reviewed. As different tests were used for children with different ages and language skills, children were grouped into categories according to the level of postoperative speech perception benefit. RESULTS: The results showed that children in the higher categories of aided preoperative residual hearing showed significant scores on open-set word and sentence perception tests using the implant alone. For children in lower categories of aided residual hearing, results were variable within the groups. More than 90% of children with implants with aided residual hearing thresholds in the speech range above 1 kHz achieved open-set understanding of words and sentences. CONCLUSION: While the results of this preliminary study confirm previous findings of differential outcomes for children with different levels of preoperative residual hearing, they suggest that children with severe to profound hearing impairments should be considered for cochlear implantation. PMID- 9391630 TI - Profiling linguistic outcomes in young children after cochlear implantation. AB - OBJECTIVE: To summarize the results of cost-utility analyses of pediatric cochlear implantation (PCI) in the United Kingdom. METHOD: Analyses were based on measured costs of health care and measured educational placements, but on estimates of the gain utility which results from PCI and estimates of the costs of educational placements. RESULTS: The cost-utility ratio calculated from the costs of health care falls on the margin of the range considered acceptable within the British health-care system. If estimates of cost-savings associated with educational placements are also considered, the resulting ratio is similar to that of other therapies provided within the British health-care system. CONCLUSION: The analysis is highly sensitive to assumptions about future costs and benefits. There is a need to reduce the number of assumptions by measuring those values which are currently estimated: in particular, the gain in utility associated with PCI and the costs of different educational placements. PMID- 9391631 TI - Long-term speech perception in children with cochlear implants compared with children with conventional hearing aids. AB - OBJECTIVE: To determine the speech perception of children with cochlear implants. SUBJECTS AND METHODS: Speech perception results of seven children with cochlear implants (excellent performers), who showed stable speech recognition scores in the long term, were compared with those of severely hearing-impaired children with conventional hearing aids (reference group). The groups of children were matched according to their mean free-field aided thresholds. RESULTS: The results of the open-set word recognition test were comparable in the two groups. CONCLUSION: If we consider the results of the hearing aid users as the gold standard, the results suggest that speech recognition in selected children with a cochlear implant is close to optimal. PMID- 9391632 TI - Some aspects of language development in normal-hearing children and children with cochlear implants. AB - OBJECTIVE: This article presents some important processes of normal child language acquisition and applies them to language acquisition data of children with cochlear implants. DATA SOURCES: Modern studies of language acquisition, covering various languages, have demonstrated a close link between linguistic and cognitive development. Sensorimotor intelligence provides a construction of reality on which the first grammatical structures are built, encoding a number of relations which hold between objects, persons, events, and localizations. When acquiring the more complex morphological and syntactic aspects of their mother tongue, children use a number of characteristic information processing strategies which make some formal markings easier to learn than others. There is considerable variability across children with respect to rate of acquisition, the use of imitation, and analytic versus holistic processing strategies. Caregivers' language input can facilitate language acquisition, notably the use of expansions and reformulations, and a generally accepting style. EMPIRICAL STUDY OF CHILDREN WITH COCHLEAR IMPLANTS: Language acquisition data from two children with cochlear implants show great differences with respect to rate of acquisition, construction of the German case system, and syntax. Whereas one child discovers the regularities of the case inflectional system quickly, the other child appears to prefer holistic and rote learning processes and uses a sequential strategy for combining words. It is suggested that variability between children with cochlear implants may be due to different frequencies of actually processed linguistic items. CONCLUSIONS: Future research should compare language development in children with cochlear implants and those with normal hearing making use of psycholinguistic methods of research design and analysis. PMID- 9391634 TI - Speech perception performance of congenitally deaf patients with a cochlear implant: the effect of age at implantation. AB - The relation between age at cochlear implantation and long-term open-set speech recognition was studied in a group of nine congenitally deaf children. The age at cochlear implant surgery ranged from 4 to 13 years. The results showed that there was a tendency toward poorer results in the children implanted at a relatively older age. However, the results also indicated that an upper limit for age at implantation cannot yet be defined in these children. PMID- 9391633 TI - The effect of language knowledge on speech perception: what are we really assessing? AB - OBJECTIVE: The authors examined whether open-set speech perception scores are limited by knowledge of vocabulary and syntax and further considered whether remediation of vocabulary and syntax will increase open-set speech perception scores. STUDY DESIGN: This was a repeated-measures study design in the setting of a primary (elementary) school for the hearing-impaired. PATIENTS: The study population was composed of three hearing-impaired children using Nucleus 22 channel cochlear implant. INTERVENTION: Intervention used was language remediation sessions. MAIN OUTCOME MEASURES: The main outcome measures were assessment of auditory-alone speech perception benefit using open-set words and sentences and assessment of syntactic knowledge using the Test of Syntactic Ability. Outcome measures were applied before and after remediation. RESULTS: Child 1 and child 2 showed a significant postremediation improvement in their overall scores on the Test of Syntactic Ability and in their ability to perceive words learned during remediation. Child 1 and child 2 also showed a significant improvement in their scores on a modified Bamford-Kowal-Bench open-set sentence test, which specifically targeted grammatical constructs trained in remediation sessions. CONCLUSIONS: Remediation of language knowledge deficits significantly improved open-set speech perception for two children, suggesting a need to include language remediation in cochlear implant habilitation programs. PMID- 9391635 TI - Speech perception in noise with implant and hearing aid. AB - OBJECTIVE: To compare the perception of speech in quiet and in noise by adults using a cochlear implant on its own or a cochlear implant and hearing aid together. STUDY DESIGN: Repeated measures. SETTING: Laboratory study using subjects' own speech processors. PATIENTS: Two groups of cochlear implant users (Australian and American) with some residual hearing in the non-implanted ear (pure tone average thresholds at 500 Hz, 1 kHz, and 2 kHz of 75-112 dB HL). INTERVENTION(S): Conventional hearing aid and cochlear implant in opposite ears. MAIN OUTCOME MEASURES: Speech perception was evaluated using recorded lists of CUNY sentences and lists of CNC words in quiet and in background noise. RESULTS: Speech scores were significantly higher with implant and hearing aid together compared to implant alone. The binaural advantage was greater in background noise than it was in quiet for CUNY sentences in the American listeners. CONCLUSION: Severely-to-profoundly hearing impaired adults may benefit from combined fitting of implants and conventional hearing aids in opposite ears. PMID- 9391636 TI - Auditory perceptual abilities in a child with a nucleus and a Clarion cochlear implant. AB - This case study will review the performance of a 7-year-old female who was implanted at the age of 3 years 9 months with a Nucleus 22 channel device. This child was deafened from pneumococcal meningis at the age of 8 months and was placed in an early intervention program which uses simultaneous communication (i.e. speech with sign language). The teaching staff of this particular educational setting has collaborated closely with the Cochlear Implant Center at Manhattan Eye, Ear and Throat Hospital and the team's teacher of the deaf. The child utilized the implant on a daily basis for a period of 2 years 10 months. Performance on a variety of auditory perceptual tests were obtained at 1 year and two year intervals. After almost 3 years of implant use, the child suffered an internal receiver failure. The Nucleus device was explanted and the child was implanted with a Clarion Cochlear Implant System. Performance on a similar set of auditory perceptual tests obtained after 3 and 6 months use indicated better performance with the Clarion device. In addition to the scores on the individual tests, a comparison questionnaire was used to obtain impressions from the parents and the school personnel. Results should be reviewed with caution since this study investigates the responses of a single child who uses each of these two different implants and cannot be generalized. PMID- 9391637 TI - Performance of the new Clarion speech processor 1.2 in quiet and in noise. AB - OBJECTIVE: To evaluate the benefit of the new Clarion speech processor 1.2. STUDY DESIGN: Fifty-two subjects who received a Clarion cochlear implant during 1993 and 1995 were upgraded with the new Clarion speech processor 1.2. All subjects were tested with a speech test battery in quiet and in noise, first with the 1.0 processor and 1 month later with the new 1.2 processor. To avoid ceiling or floor effects, the subjects were divided into three groups according to their test results with the Freiburger Monosyllabic Word Test (group A < 10%, group B 10 50%, and group C > 50%). RESULTS: The results indicated that all subjects had an improvement with the new speech processor. This improvement was statistically significant (p < 0.05) for group C in all conditions, and for group B in the tests with noise. CONCLUSION: While the better and good performers improved their test scores significantly, low performers seemed to derive little benefit from the technical improvements of the same speech processing strategy. Perhaps this group might gain more from a different processing scheme. PMID- 9391638 TI - Multichannel cochlear implantation in postmeningitic and congenitally deaf children. AB - OBJECTIVE: To test the view that prelinguistic postmeningitic deaf (PMD) children outperform congenitally deaf children (CD) in the first year following cochlear implantation. STUDY DESIGN AND PATIENTS: We evaluated 85 children with ages (at implantation) ranging from 1.9 years to 13.5 years (mean age 5.4 years). The Listening Progress scale was used to assess the developing use of audition 3, 6, and 12 months after implantation. RESULTS: In contrast to previous reports, the PMD children achieved statistically significantly lower scores than CD children. PMID- 9391639 TI - Some experiences with the nucleus 20 + 2 cochlear implant in adults and children. AB - OBJECTIVE: To evaluate the effectiveness of the Nucleus 20 + 2 implant in a group of patients. PATIENTS: Fifteen children and 11 adults who have received the Nucleus 20 + 2 implant since late 1993. RESULTS: The outcome in most patients has been favorable. PMID- 9391640 TI - Telephone speech comprehension in children with multichannel cochlear implants. AB - Telephone speech comprehension is being evaluated in six prelingually deaf children implanted with the Nucleus 22 prosthesis fitted with the Speak strategy. All of them have had at least 1.5 years of experience with their implant. When the tests began, they had already had at least 2 months' experience with the same map in their speech processor. The children were trained in the use of the telephone as part of the rehabilitation program. None of them used it regularly but as a game that they found very entertaining. A special battery, the Bate-fon (bateria para telefono = telephone battery), was designed for training and evaluation purposes. It includes the five Spanish vowels in isolation, diphthongs, onomatopoetic animal voices, two-syllable, and three-syllable words. The tests were administered 1.5-2 years after the switch-on of their speech processor. Standard acoustic telephone coupling was used. The speech material was presented to the child on colored cards. Stimuli were presented twice. Children were informed when the response was incorrect. Averaged results indicated that the percentages of correct responses for all the speech material increase in the second presentation. All children have shown some degree of telephone communication abilities. As a result of the training, some of the children are using the telephone to communicate with their families. PMID- 9391641 TI - Speech recognition performance of pediatric Clarion patients. AB - OBJECTIVE: To evaluate the speech performance of children with the Clarion Multi Strategy Cochlear Implant. PATIENTS: Prelingually deafened children who had received the Clarion implant. METHODS: The Spondee and Monosyllable Word Identification subtest of the Early Speech Perception test, the Glendonald Auditory Screening Procedure (GASP), and the Phonetically Balanced Kindergarten test (PB-K) were administered to subjects 3 and 6 months after implantation. RESULTS: At 3 months, subjects' performance was higher than preoperatively with hearing aids. At 6 months, performance improved further. Scores were higher on the ESP and GASP than on the PB-K. The scores indicate better levels of speech recognition than were obtained with older implant processing strategies. Subjects varied considerably in their performance. The results are preliminary because of the small sample. PMID- 9391642 TI - Speech recognition in children with hearing aids versus results in children with implants. PMID- 9391643 TI - Performance of 2- and 3-year-old children and prediction of 4-year from 1-year performance. AB - OBJECTIVE: To examine whether children perform better when they receive cochlear implants when they are 2 to 4 years of age than when they are older, and to determine whether 4-year performance can be predicted from 1-year results. METHOD: Children in two age groups (2 to 4, 4 to 9 years) were tested for performance, and the age groups were compared. Children were also tested 1 and 4 years after implantation. RESULTS: The results suggest that the "implanted young" group scored higher than the "implanted old" group after 36 months, and that 1 year performance is helpful in predicting 4-year performance. CONCLUSION: It may be desirable for children to undergo implantation when they are under 2 years of age, provided that appropriate selection criteria can be determined. PMID- 9391644 TI - Initial results from the clinical trial of the nucleus 21-channel auditory brain stem implant. PMID- 9391645 TI - Stress experienced by parents of children with cochlear implants compared with parents of deaf children and hearing children. AB - OBJECTIVE: To compare the stress experienced by parents of children with cochlear implants with that experienced by parents of deaf children and hearing children. STUDY DESIGN: The Parenting Stress Index and problem-oriented interviews were used with the parents of these three groups of children. RESULTS: Parents of deaf children were found to experience greater levels of stress than parents in the other two groups. CONCLUSION: Parents of children with cochlear implants experience about the same level of stress as parents of hearing children. PMID- 9391646 TI - Comparative study of English- and German-speaking children with the Clarion cochlear implant. AB - OBJECTIVE: To compare the performance of English-speaking and German-speaking children with Clarion cochlear implants on the Meaningful Auditory Integration Scale (MAIS). SUBJECTS AND METHODS: Prelingually deafened German-speaking children and English-speaking children with Clarion cochlear implants were assessed on the MAIS pre- and postoperatively. Data were analyzed in terms of the absolute score, preoperatively and at two postoperative intervals, and the improvement score. RESULTS AND DISCUSSION: Preoperatively, the MAIS scores of German-speaking children were slightly lower than those of English-speaking children. Postoperatively, this difference became more pronounced. Substantial deviation within each group of subjects was suggested by the size of the standard deviations. PMID- 9391647 TI - Cost-effectiveness considerations in pediatric cochlear implantation. AB - OBJECTIVE: To summarizes the results of cost-utility analyses of pediatric cochlear implantation (CI) in the United Kingdom. METHOD: Analysis is based on the direct costs of medical and rehabilitative management and also on emerging evidence that implantation leads to a shift in educational placements in favor of mainstreaming with support. RESULT: The resulting cost-utility ratio falls on the margin of the range considered acceptable within the British health-care system. The analysis also suggests that pediatric CI could be acceptably cost-effective. CONCLUSION: The next step should be to measure the costs of alternative educational settings directly. PMID- 9391648 TI - Preliminary conversation with the parents before cochlear implantation. AB - OBJECTIVE: To provide parents of hearing-impaired children about to undergo cochlear implantation with an opportunity to discuss their expectations and the pedagogical implications of surgery. SETTING: Parents and children participate in discussions at the medical center where the cochlear implantation will take place. PATIENTS: More than 700 children and their parents have participated. OUTCOMES: Parents of congenitally deaf children differ somewhat in their expectations and behavior from the parents of postlingually deafened children. PMID- 9391649 TI - Nucleoprotein complexes harboring an extrachromosomal DNA closely related to 7 S DNA of avian myeloblastosis virus: physico-chemical properties and representation of nucleic acids. AB - The source of avian myeloblastosis virus (AMV) DNA, an extrachromosomal small polydisperse DNA, present in the material forming the postmicrosomal sediment (POMS) of lysed chicken leukemic myeloblasts (CHLMs) is organized into nucleoprotein (NP) complexes containing always RNA. This material, radioactively double-labelled for DNA and RNA, separated in isopycnic sucrose gradients into three POMS components (A,B,C) differing from one another in properties of labelling for DNA and RNA, sucrose densities (1.21, 1.18 and 1.08 g/cm3 for components A, B and C, respectively) and sedimentation properties of NP complexes which constituted the individual POMS components. The NP complexes present in representative fractions of POMS components A, B and C sedimented at 37.3 and 27.3, at 15.3 and 7.4, and at 11.3 and 5.5, respectively. They differed also in the length of DNAs they were harboring. Radioactively double-labelled nucleic acids (NAs) of POMS components A, B and C sedimented at 9, 7 and 3.5 S, respectively, and the sedimentation characteristics of both labels corresponded with those significant for replication intermediates. Electrophoretic characteristics of these NAs indicated that we dealt with DNA and RNA products of a lagging DNA strand synthesis (LSS) taking place, evidently, on pieces of the lagging sites of replicating DNA strands that were cut out at predicted sites by nucleases. As regards the origin of AMV DNA, we show that the major and minor portions of this DNA might be descending from NAs harbored in NP complexes of POMS components B and C, respectively, pointing out selectivity of segregation of this DNA from the cell into AMV core. PMID- 9391650 TI - Activities of a lagging DNA strand synthesis of nucleoprotein complexes harboring an extrachromosomal DNA closely related to avian myeloblastosis virus core-bound DNA. AB - Nucleoprotein (NP) complexes constituting the material of the postmicrosomal sediment (POMS) and its three basic components (A, B, C) (Riman and Sulova, 1997a), harboring an extrachromosomal DNA closely related to AMV DNA were found to possess DNA- and RNA-synthesizing activities (SAs) reflecting the ability of this material to be intensely labelled for DNA and RNA, respectively. The types of these NA-SAs were compatible with those significant for a lagging DNA strand synthesis (LSS). The use of selective inhibitors and of the proliferating cell nuclear antigen (PCNA) disclosed a successive involvement of alpha DNA polymerase (pol) and PCNA-insensitive delta DNA pol in LSS. In this respect, we show gradual changes in the representation of activities (As) of both mentioned DNA pols in the NP complexes of the individual POMS components. Those of POMS component C contained alpha DNA pol As only, while a distinct portion of DNA SAs of POMS component B was represented on expense of alpha DNA pol As by PCNA-insensitive delta DNA pol (epsilon DNA pol), As which represented practically all the DNA SAs of POMS component A. The type of RNA SAs of this material represented mostly by primase (Pr) As corresponded well with the nature of LSS. An exception was represented by a minor portion of RNA-SAs of POMS component A which was alpha amanitin-sensitive like RNA pol II. Moreover, analyzing this natural model replication system, we found that the carbonyldiphosphonate (COMDP), a selective inhibitor of the PCNA-insensitive delta DNA pol, was a strong activator of Pr-As and/or Pr-alpha DNA pol As of NP complexes of POMS component C. PMID- 9391651 TI - Products of a lagging DNA strand synthesis of nucleoprotein complexes harboring an extrachromosomal DNA closely related to avian myeloblastosis virus core-bound DNA. AB - Nucleoprotein (NP) complexes present in selected fractions of the separated three basic components A, B, C of the postmicrosomal sediment (POMS) (Riman and Sulova, 1997a) were used as a source of nucleic acid synthesizing activities (NA-SAs) expressed in reactions in vitro. These were performed in the absence and presence of N2-(p-butylphenyl) deoxyguanosine 5'-triphosphate (BuPdGTP), aphidicolin (Aph) and carbonyldiphosphonate (COMDP), inhibitors allowing differentiation between two DNA polymerases (pols) involved in a lagging DNA strand synthesis (LSS). Reaction products were isolated and analyzed by polyacrylamide gel electrophoresis (PAGE) in denaturing conditions. Products labelled with [alpha 32P]dAMP or [alpha-32P]AMP were represented by intermediates significant for LSS. Okazaki fragment precursors, whose synthesis was inhibited by BuPdGTP and resistant to Aph, and whose radioactive RNA label was DNase I-sensitive, represented products formed in vitro by NP complexes of POMS component C. Okazaki fragments 127 b in length, whose synthesis was insensitive to BuPdGTP but inhibited by Aph and COMDP, were characteristic of the reactions accomplished by NP complexes of POMS component B while products of NA-SAs of NP complexes of POMS component A were represented by Okazaki fragments up to 280 b in length, whose synthesis was most sensitive to Aph. In accord with previous data (Riman and Sulova, 1997b), COMDP strongly stimulated production of RNAs corresponding in length with initiator RNAs (iRNAs). However, in dependence on reaction conditions also ribodeoxyribonucleotide primers can be produced by NP complexes of POMS component C, suggesting the occurrence of two primase (Pr) catalytic modes influenced by dNTP/rNTP relation and thus, by COMDP, a strong competitor for dNTPs. These results represent the first direct evidence that an extrachromosomal DNA organized into special NP complexes can be replicated extrachromosomally by a mechanism of LSS. PMID- 9391652 TI - Augmentation of natural killer cell activity induced by cytomegalovirus infection in mice treated with FK506. AB - Comparable rates of patient and graft survival after FK506 and cyclosporine treatments have been reported in the prevention of liver allograft rejection. On this basis, we examined the effect of FK506 on pathogenesis of cytomegalovirus (CMV) infection in mice. FK506 induced apparent immunosuppression in mice which could be monitored by the level of antibody production. The effective dose of trinitrophenyl-keyhole limpet hemocyanin (TNP-KLH) for 50% reduction in antibody production was 0.9 mg/kg. Even in such an immunosuppressed status at this or higher dose of FK506, CMV infection was relatively alleviated, which was observed by the frequency of virus isolation and the mean virus titer of the lungs of mice treated with 0.1-1 mg/kg FK506 in comparison to untreated mice. The dose of FK506 attaining 50% frequency of lung infection was 1.5 mg/kg. The activity of natural killer (NK) cells was enhanced in infected mice. This enhancement was stronger in infected mice treated with FK506 at 0.32 mg/kg and 10 mg/kg than in untreated infected mice on day 3 post infection (p.i.). Thus, an immunosuppressant FK506 augmented inducible NK cell activity and alleviated MCMV infection even under immunosuppression. PMID- 9391653 TI - Electronmicroscopic study of human herpesvirus 6-infected human T cell lines superinfected with human immunodeficiency virus type 1. AB - Human herpesvirus 6 (HHV-6) has been proposed as one of the co-factors responsible for the development of acquired immunodeficiency syndrome (AIDS) in human immunodeficiency virus (HIV) carriers. We analyzed the interaction between HHV-6 and HIV-1 in superinfected cells. Cell-free HIV-1 could superinfect human T cell lines, MT-4 and Molt-4, which had been previously infected with HHV-6. Both HHV-1 and HHV-6 replicated in the same cells. We observed two types of morphologically distinguished cells as early as 4 days after superinfection. One type (D) was degenerate cells with intracellular and extracellular HHV-6 and with less HIV-1 virions. The other type (I) was relatively intact cells with both HIV 1 and HHV-6 virions. Replication of HIV-1 was more active in the type I as compared with type D cells. The level of HIV-1 reverse transcriptase (RT) activity in the culture supernatants of cells superinfected on day 0 declined after day 7, while that in the supernatants of cell cultures infected with HIV-1 alone remained high between days 12 and 40. These results suggest that the superinfection of the HHV-6-infected cells with HIV-1 may induce a degenerative process in these cells. PMID- 9391654 TI - Evolutionary stasis of M1 gene of human influenza A viruses and the possibility of their subtyping by restriction analysis of M1 gene polymerase chain reaction product. AB - Nucleotide (nt) and amino acid (aa) sequences of the M1 protein in 36 human influenza A viruses were analyzed. The neighbor joining tree of the nt sequences revealed several lineages associated with past epidemics of human influenza. However, the tree of aa sequences revealed only few specific lineages. This discrepancy in phylogeny between nt and aa sequences indicates that the M1 protein of human influenza A virus nearly reached an evolutionary stasis. A simple subtyping method of human influenza A viruses by restriction fragment length polymorphism (RFLP) analysis of M1 gene polymerase chain reaction (PCR) products is discussed. PMID- 9391655 TI - Detection of strawberry vein banding virus by polymerase chain reaction and dot blot hybridization. AB - Strawberry vein banding virus (SVBV) is one of seventeen members of the family Caulimoviridae. Natural infection with the virus is known in Fragaria species only. Infections caused by SVBV are often symptomless (1), but their significance increases in mixed infections with strawberry crinkle or strawberry latent C viruses (2,3). This virus has been originally found on strawberries in USA and firstly described by Frazier (4), but it is probably world-wide distributed by planting or breeding materials. SVBV has been observed on cultivated strawberries in North America, Australia, Brazil, Japan (5) and recently in Europe (6,7). The concentration of SVBV in infected plants is usually very low. Its detection by ELISA is impossible because of lack of specific antibodies. Evidence of the caulimovirus nature of SVBV has been confirmed by its circular dsDNA genome, shape and size of viral particles (8), presence of cytoplasmic inclusion bodies typical for caulimoviruses, and distant serological relationship with cauliflower mosaic virus (CaMV, 9). In this paper we present detection of SVBV by combination of two detection methods--polymerase chain reaction (PCR) and dot blot hybridization with a non-radioactive probe. PMID- 9391656 TI - Gay youth and their precautionary sexual behaviors: the Sydney men and sexual health study. AB - Many commentators have positioned Western gay youth as a high-risk group for HIV infection and obscured important cultural, social, and contextual differences between populations. This study compares risk of HIV transmission factors among 216 young (under 25 years) and 822 older (25 years or over) homosexually active men, recruited through Sydney gay community and other sources. Bivariate and multivariate analyses of survey data that were collected by personal interviews consistently supported our hypothesis of no difference in HIV-related risk factors between young and older men. Although young men in this cohort were more likely to be of unknown serostatus, they were at least as knowledgeable, as attached to gay community, and as precautionary in their sexual behaviors with regular and casual male partners as their older counterparts. Safety campaigns targeting these young gay men should focus, for example, on their lower rates of HIV antibody testing and not be based on a false premise of hedonistic, uninformed, and disenfranchised youth. PMID- 9391657 TI - Condom use by Dutch men with commercial heterosexual contacts: determinants and considerations. AB - We report responses from 559 clients of female prostitutes, with a view to determining to what extent previously identified factors play a part in condom use. To increase the response rate to advertisements in daily and weekly newspapers, interviews were held by phone. This procedure had the advantage of ensuring the anonymity many clients demanded. Of those clients having vaginal or anal contact (91%), 14% had not always used condoms in the previous year. Compared with consistent condom users, these men were less highly educated, had twice as many commercial contacts, and had more contacts with "steady" prostitutes. They were either more emotionally motivated to visit prostitutes than were consistent condom users or exhibited a stronger need for sexual variation. They showed a more compulsive attitude toward visiting prostitutes, had a more negative attitude toward prostitution in general, evaluated condoms more negatively, had a higher personal efficacy to achieve unsafe contacts, and had a higher general risk assessment, commensurate with their behavior. Men with only safe contacts had either an intrinsic or an extrinsic motivation for condom use. Among extrinsically motivated men, their behavior change was more recent and had not yet taken root: They still envisioned unsafe commercial sex to be possible in the future. Education aimed at the small group of men practicing unsafe contacts will not easily and directly lead to behavior change. But these educational activities may support prostitutes to persist in (consistent) condom use, regardless of clients' pressure to do otherwise. PMID- 9391658 TI - Attitudes, norms, self-efficacy, and stage of change among out-of-treatment female crack cocaine users: a pilot study. AB - The level of sexual risk among crack cocaine uses had remained high, regardless of their level of AIDS knowledge. Consequently, researchers have advocated the use of rigorous behavioral theory in aiding epidemiological research and intervention. To test whether stage of change for "insisting that men (besides your main partner) use condoms every time you have sex with them" was associated with behavioral attitudes, subjective norms, and self-efficacy among female crack cocaine users, a behavioral context questionnaire was administered to 61 female crack cocaine users who were recruited in the field and interviewed in an urban HIV testing center. Results indicated that all three were associated with stage of change and that self-efficacy was the variable most strongly related to stage of change. PMID- 9391659 TI - Psychosocial antecedents of needle/syringe disinfection by drug users: a theory based prospective analysis. AB - Working from the AIDS risk reduction model and other theories of behavior change, we tested psychosocial antecedents of needle/syringe disinfection by 136 injection drug users. High perceived self-efficacy for risk reduction exerted a positive effect on needles/syringe disinfection attempts 1 year later. Self efficacy was, in turn, related to lower perceived infection risk, peer norms more favorable to risk reduction, and greater knowledge of AIDS. Behavioral intention had no significant effect on subsequent disinfection attempts. These results suggest that disinfecting needles/syringes is partly non-volitional; that high perceived infection risk may be counterproductive to injection risk reduction; and that perceive self-efficacy, but not intention to change behavior, may be a useful leverage point for AIDS preventive intervention. PMID- 9391660 TI - AIDS knowledge and risk perception of cocaine and crack users in a national household survey. AB - Awareness of AIDS among cocaine and crack users has never been studied using national data representative of the U.S. household population. Data from the 1991 National Health Interview Survey were analyzed. Respondents who reported cocaine (n = 448) or crack use (n = 100) in the past year were compared with those who reported never using any form of cocaine (n = 17,259). AIDS knowledge, HIV testing, risk behavior, and perceived risk for HIV were outcomes studied. Over 96% of the drug users know the term HIV compared with 89% of the nonusers. A higher proportion of cocaine users reorganized the effectiveness of condoms compared with nonusers (93% vs. 84%). Over 96% of all groups knew the risk of sharing needles. Cocaine and crack users were more likely to have been tested for HIV (27% and 28%) compared with nonusers (19%), yet less than one third of those tested actually received HIV counseling. High-risk behavior was acknowledged by 22% of cocaine users and 33% of crack users. However, only 10% and 14% respectively considered themselves to be at increased risk for having or getting HIV. These data suggest that cocaine and crack users are knowledgeable regarding HIV/AIDS, however they are underestimating their real risk of infection with HIV. PMID- 9391661 TI - AIDS and condoms in Brasilia: a telephone survey. AB - A telephone survey was conducted to measure AIDS knowledge, media usage and condom attitudes and behaviors among 500 adults aged 18 to 49 in Brasilia, as well as to evaluate the feasibility of the telephone survey method in a developing country. The response rate was 91.6%. Respondents had good knowledge about correct modes of HIV transmission and prevention but also believed HIV was transmitted through blood donation, public toilets, swimming pools, and mosquito bites. TV and newspapers were the most important sources of information on health matters and AIDS, though health workers were considered the most credible sources of such information. Only 19% of sexual encounters in the 4 weeks prior to the survey included condoms. Single and younger respondents and those with more positive attitudes used condoms more frequently. More work is needed to identify appropriate messages to motivate people to use condoms. Telephone surveys regarding AIDS and sexual attitudes and behaviors are feasible in Brasilia, a planned community with universal telephone coverage. PMID- 9391662 TI - Perspectives on the future of temporal bone research. PMID- 9391663 TI - Labyrinthine fistula as a complication of cholesteatoma. AB - HYPOTHESIS: The objective of this study was to present the authors' experience in the management of labyrinthine fistula caused by cholesteatoma. METHODS: The clinical charts of 92 patients who underwent surgical procedures for cholesteatoma complicated by labyrinthine fistula between 1979 and 1975 were reviewed retrospectively. In this period, 1,205 patients were operated on for cholesteatoma. In each patient, the site and size of the fistula were evaluated during surgery and the hearing thresholds were compared before and after surgery. RESULTS: The fistula involved the lateral semicircular canal in 71 patients. Multiple fistulas were observed in nine patients. Postoperative hearing levels were unchanged or improved in 83.7% of patients. Comparison between hearing outcomes and size of the fistula showed better findings when smaller size fistulas were found. No significant differences between open and closed techniques were detected. Favorable outcomes were obtained in patients treated with surgical obliteration of the interrupted labyrinth. CONCLUSIONS: The current study confirmed that careful manipulation of the labyrinthine fistula is mandatory to preserve hearing functions for these patients. According to the authors' experience, the future trend for fistula treatment could be directed toward less conservative techniques compared with the previous indications favoring methods of interruption and subsequent obliteration of the semicircular canals. PMID- 9391664 TI - Stimulation of epithelial healing in chronic postoperative otorrhea using lyophilized cultured keratinocyte lysates. AB - OBJECTIVE: After tympanoplasty, despite a closed tympanic graft, some patients continue to have persistent otorrhea due to insufficient epithelial healing and granulation tissue formation in the depths of the outer ear canal. When all medical therapies fail, many otologists undertake revision surgery, usually with free skin grafting. To avoid surgery, the authors sought to improve this condition with a lysate of lyophilized cultured allogeneic keratinocytes. STUDY DESIGN AND PATIENTS: In this prospective pilot study, lyophilized cultured allogeneic keratinocyte lysates have been administered in 27 patients. These patients had uncontrollable otorrhea that resisted medical (topical) therapy for at least 6 months. MAIN OUTCOME MEASURE: The criterion of success was a complete epithelialization and cessation of otorrhea. RESULTS: After an average of 2 applications, cessation of otorrhea was achieved in 20 cases (74%). Three patients (11%) relapsed after 3 months. The other ears (63%) still were dry at the 1-year final evaluation. CONCLUSIONS: These results are similar to those obtained after application of sheets of viable cultured keratinocytes of autologous as well as of allogeneic origin. Because the soluble lysate can be incorporated into ototopical drops, the lysate technique is more "user-friendly" and can be applicable in any outpatient clinic. Because keratinocytes contain many growth factors (e.g., epidermal growth factor, basic fibroblast growth factor, platelet-derived growth factor, transforming growth factor), the authors speculate that the release of those intracellular growth factors is responsible for the observed therapeutic effect. This form of therapy by its combination of several growth factors might be considered a more physiologic method than the, also still experimental, growth factor therapy in which high doses of only single growth factor are used. PMID- 9391665 TI - Intra-operative electrocochleography in stapedectomy and ossicular reconstruction. AB - OBJECTIVE: The purpose of this study was to evaluate the effectiveness of intraoperative electrocochleography (ECOG) in predicating the postoperative hearing improvement in surgery for conductive hearing loss. STUDY DESIGN: This study was a prospective study of 22 patients undergoing intraoperative electrocochleography during a stapedectomy. SETTING: The study was performed in a tertiary referral center. PATIENTS: Intraoperative electrocochleography was performed in 22 patients 27-73 years of age undergoing a stapedectomy for otosclerosis under general anesthesia. INTERVENTION: For each patient, the N1 threshold to click stimulation was measured intraoperatively, before and after the reconstruction. MAIN OUTCOME MEASURES: The intraoperative ECOG thresholds were compared with the pre and postoperative audiograms. RESULTS: Postreconstruction ECOG's demonstrated improvements in the N1 threshold in 19 cases, and were unchanged in 1 case. In each of these cases, improvement in the intraoperative N1 threshold corresponded with improvement in the postoperative audiogram compared with the preoperative studies. In two other cases the postreconstruction ECOG was nearly unobtainable, despite improved hearing postoperatively. CONCLUSION: Intraoperative ECOG appears to be an effective tool for verifying the functional integrity of ossicular reconstructions as in stapedectomies. We speculate that intraoperative ECOG may allow the surgeon to "fine tune" the reconstruction to optimize the hearing results. PMID- 9391666 TI - Cost analysis of cochlear implants in deaf children in The Netherlands. AB - OBJECTIVE: The purpose of the study was to determine the costs of cochlear implants in children regarding the phases of selection, implantation, rehabilitation, and aftercare. STUDY DESIGN: This study was a prospective cost analysis paralleling a noncomparative observational study. SETTING: This study was conducted at a university hospital to evaluate cost data on selection and implantation and at an institute for the deaf to evaluate cost data on rehabilitation and aftercare. PATIENTS: The study group consisted of prelingual deaf children (mean age, 7 years; range, 4-11 years). INTERVENTION: A total of 106 deaf children were screened, of whom 20 received a cochlear implant. MAIN OUTCOME MEASURES: This study concentrated on the cost of cochlear implants. Volumes of utilization of human resources and materials were registered during the 1-year follow-up. For the subsequent period, volumes were modeled on planned aftercare activities. RESULTS: Real total medical costs per implanted child were $63,922; selection phase, $7,747; implantation phase, $30,442; rehabilitation phase, $13,428; and aftercare, $12,305. Nonmedical costs were $1,839. Calculations were based on 1994 prices, and a time horizon of 5 years was used. The economic consequences of cochlear implants on educational needs were not taken into account because of the limited follow-up period. A sensitivity analysis of the rate of implanted children as part of the number of screened children showed a moderate impact on the total cost. CONCLUSIONS: Compared to the results of cost analysis in other countries, the costs of the pediatric cochlear implants program in The Netherlands are relatively high. Most discrepancies can be explained by methodologic differences in the cost analyses. PMID- 9391667 TI - Clarion cochlear implant: short-term effects on voice parameters. AB - OBJECTIVE: This study aimed to evaluate the moment-to-moment auditory control of voice at an early stage after implantation with Clarion cochlear implants. STUDY DESIGN: A perceptive and electroacoustic evaluation of the voice was carried out through a digital analysis immediately after the activation of the implant, before the fitting procedure has begun. SETTING: The study was performed at the Department of Otolaryngology, University "La Sapienza" of Rome. PATIENTS: Nine profoundly deaf subjects (five post-linguistic deaf adults, two pre-linguistic deaf children and two peri-linguistic deaf subjects, one adult and one child). INTERVENTION: Surgical insertion of a Clarion cochlear device. MAIN OUTCOME MEASURES: Qualitative (short-term pitch and energy perturbation, intonation, vocal attack, quality, and intensity) and quantitative (F0, F1, F2 and F3 frequency values), under non activated (NAI) and activated (AI) condition, have been obtained. RESULTS: In the majority of patients, the perceptive evaluation under AI showed a lowering of voice intonation, a better control of voice intensity, and a reduction of nasal quality. These findings were confirmed by a significant lowering of F0 (Wicoxon non parametric test) in all cases and lowering of F1 and F2 in five cases. Additionally, a better definition of all formats in the majority of cases as well as by a parallelism of pitch and energy profile was observed. CONCLUSIONS: The Clarion cochlear implant device provided a recognizable moment-to-moment auditory control on voice and articulatory patterns. By monitoring the articulated voice during adjustment of the electrical stimulation at the first fitting session, one may be able to include these data and assist in the selection of the best rehabilitative strategy. PMID- 9391668 TI - Immediate effects of middle ear pressure changes on the electrocochleographic recordings in patients with Meniere's disease: a clinical placebo-controlled study. AB - OBJECTIVE: The aim of the study was to evaluate effects of middle ear pressure changes on the electrocochleographic responses in patients with well-defined Meniere's disease. STUDY DESIGN AND INTERVENTIONS: The investigation was conducted as a placebo-controlled, randomized clinical study. Electrocochleographic measurements were performed before and after the insertion of a transtympanic ventilation tube and immediately after the exposure to active or placebo treatment. SETTING: The study was carried out in one secondary referral center and one tertiary referral center on an ambulatory basis. PATIENTS: Thirty-nine patients with well-defined Meniere's disease were included in the study. MAIN OUTCOME MEASURES: The summating potential/action potential ratio of the electrocochleographic response complex was chosen as the main variable for statistical evaluation of results. Other parameters of the recordings such as responses to low-frequency burst stimulation also were evaluated. Subjective symptoms (e.g., vertigo, tinnitus, and aural pressure) were assessed before and after insertion of the ventilation tube and before and after exposure to treatment. RESULTS: A statistical difference was shown in the electrocochleographic response in the active group before and after exposure to middle ear pressure changes. In the placebo group, no change was found. Changes in electrocochleographic parameters in the active group indicated an improvement in inner ear electrophysiology. No significant changes were found in subjective symptoms in the active or the placebo group. Evaluation before and after insertion of the ventilation tube showed no significant improvement in any variable. CONCLUSIONS: This is the first study in which electrophysiologic parameters were evaluated in a placebo-controlled clinical trial of overpressure treatment in Meniere's disease. The results show that electrophysiologic parameters can be improved by the application of positive pressure pulses of low amplitude in the middle ear. PMID- 9391669 TI - Detection of viral DNA in vestibular ganglia tissue from patients with Meniere's disease. AB - OBJECTIVE: The main goal of this study was to examine the vestibular ganglia from 11 patients with intractable classic Meniere's disease (MD) for the presence or absence of DNA from three neurotropic viruses (herpes simplex virus, cytomegalovirus, and varicella zoster virus) using exquisitely sensitive molecular biologic techniques. STUDY DESIGN: This was a prospective controlled study with vestibular ganglia from patients with MD and from patients with small vestibular schwannomas undergoing resection. Polymerase chain reaction was used for viral DNA detection from the ganglia along with known positive and negative polymerase chain reaction control subjects. SETTING: The study was performed in an academic tertiary referral center. PATIENTS: Patients for inclusion had medically uncontrolled MD, including documented fluctuating sensorineural hearing loss, episodic vertigo, and tinnitus who elected to undergo vestibular nerve section. Control patients were undergoing vestibular schwannoma removal. INTERVENTIONS: The intervention was vestibular nerve section with removal of vestibular ganglion. MAIN OUTCOME MEASURES: The presence or absence of viral DNA (herpes simplex virus, cytomegalovirus, and varicella zoster virus) in vestibular ganglion tissues detected by polymerase chain reaction. RESULTS: No viral DNA was detected in the vestibular ganglia of patients with MD (p = 0.028) nor in the control group. The likelihood of a type II or beta type error was < 10%. CONCLUSIONS: In patients with MD requiring surgical intervention, infection with herpes simplex virus, cytomegalovirus, or varicella zoster virus of the vestibular ganglia does not appear to play a major role in the pathoetiology of the disease. PMID- 9391671 TI - Role of transient-evoked otoacoustic emissions for hearing preservation in acoustic neuroma surgery. AB - OBJECTIVE: This study aimed to assess whether transient-evoked otoacoustic emissions (TEOAEs), which are known to be expressions of an intact cochlear function, could be useful for the rationale of hearing preservation in acoustic neuroma (AN) surgery. STUDY DESIGN: The TEOAEs were measured before, during, and after surgery in a consecutive series of patients affected by cerebellopontine angle tumors. SETTING: The study was performed at the Department of Otolaryngology, University "La Sapienza" of Rome. PATIENTS: Five patients with AN and one with a meningioma totally involving the Cochlear (VIII) nerve. INTERVENTION: Retrosigmoid approach on the ground of the limited AN size (within 20 mm) and the 30/70 rule, as proposed by the American Academy of Otolaryngology Head and Neck Surgery nomogram. Two patients also were selected despite a poor hearing level and the absence of TEOAEs. MAIN OUTCOME MEASURES: Preoperative and postoperative pure tone audiometry compared with TEOAEs. Intraoperative TEOAEs were compared with electrocochleographic findings. RESULTS: The TEOAEs were found to be present also in patients with AN with poor pure-tone average (PTA) threshold (i.e., > 75 dB). Intraoperatively, TEOAEs recording showed to be markedly affected by the environmental noise as well as by specific intraoperative maneuvers, such as drilling of the internal auditory canal or tumor removal or both. In the three patients in whom hearing successfully was preserved. TEOAEs were present in the first postoperative days, despite a temporary deterioration of the PTA threshold. CONCLUSIONS: The intraoperative use of TEOAEs showed to be scarcely reliable, whereas their presence in the preoperative assessment of patients with AN could lead to an extended number of patients to be selected for hearing-preservation surgery. Finally, an early postoperative identification of TEOAEs may be considered a favorable prognostic sign for foreseeing a delayed pure-tone hearing threshold recovery. PMID- 9391670 TI - Nonsurgical factors predictive of postoperative hearing for patients with vestibular schwannoma. AB - OBJECTIVE: The purposes of the study were to determine whether preoperative cochlear reserve as measured by evoked otoacoustic emissions (EOAE) as well as other hearing variables often associated with hearing preservation are correlated with hearing preservation after tumor removal and to determine whether any hearing variables are independent of tumor size as a predictor of hearing preservation. STUDY DESIGN: Preoperative audiologic data for 104 patients having vestibular nerve schwannomas removed via a retrosigmoid surgical approach were reviewed and subjected to factor analysis. SETTING: All patients were seen at the Mayo Clinic, Rochester, Minnesota. PATIENTS: The patient sample was divided into two groups based on hearing thresholds after surgery. Group I consisted of 73 ears without hearing preservation. The remaining 31 ears, group II, had preserved hearing (defined as average postoperative pure-tone thresholds < or = 85-dB HL for 0.5, 1, 2, and 3 kHz). MAIN OUTCOME MEASURES: Variables not predictive of hearing preservation were age, gender, tumor laterality, and cochlear reserve (EOAE). Variables predictive of hearing preservation were small tumor size, pure tone hearing sensitivity, speech reception thresholds, word recognition scores, integrity of cochlear nerve (acoustic reflex thresholds, and auditory brain stem response [ABR] waveforms). RESULTS: A multivariate logistic regression analysis showed that only word recognition scores at 40-dB sensation level were independent of tumor size as a predictor of hearing preservation. PMID- 9391672 TI - Radioimmunoimaging of glomus tympanicum tumors by In-111 labeled monoclonal antibody using single photon emission computed tomography. AB - OBJECTIVE: This study aimed to evaluate the diagnostic value of radioimmunoimaging by radionuclide-labeled monoclonal antibody F023C5 (MAb), raised originally against carcinoembryonic antigen (CEA), in patients with glomus tympanicum tumors. STUDY DESIGN: Prospective. SETTING: Preoperative imaging versus radioactivity of removed tumor. PATIENTS: Two patients with paraganglioma (glomus tympanicum). INTERVENTION: Diagnostic. MAIN OUTCOME MEASURE: Radiolabeled MAb accumulates in paraganglioma tissue. Single photon emission computed tomography (SPECT) provides improved detection of lesions. RESULTS: SPECT using F023C5 MAb detected the abnormal accumulation of radioactivity in the middle ear region. This method detected paraganglioma less than 1 cm in diameter. CONCLUSIONS: Successful detection of glomus tympanicum in two patients using In 111-labeled F023C5 MAb is reported. The result suggests the radioimmunoimaging using this antibody is useful for the detection of not only primary glomus tumors, but also of local recurrence and unsuspected lesion in patients with paragangliomas. PMID- 9391673 TI - Somatic neurofibromatosis type 2 gene mutations and growth characteristics in vestibular schwannoma. AB - BACKGROUND: The growth of hereditary and sporadic vestibular schwannomas shows wide variation, but what determines this is poorly understood. HYPOTHESIS: In neurofibromatosis type 2 (NF2), there is some correlation between the nature of the germline NF2 gene mutation and phenotype. Somatic mutations in the NF2 gene occur in sporadic tumors, but their relation to tumor behavior is unknown. METHODS: This study has investigated the molecular pathogenesis of vestibular schwannoma by looking for NF2 gene mutations. The authors have screened 17 exons of the NF2 gene in 91 sporadic vestibular schwannomas, 2 NF2, and 1 vagal schwannoma. These data have been correlated with a clinical growth index and a tumor cell proliferation index, determined using a monoclonal antibody to the proliferating cell nuclear antigen. RESULTS: Of the 94 tumors studied, 40 somatic gene mutations (38%) have been sequenced in 36 tumors. The mutations included 36 protein truncating mutations, 1 in-frame deletion, 2 splice site mutations, and 1 missense mutation. Regression analysis showed no correlation between the nature of the NF2 gene mutation and either the clinical (R2 = 0.006) or the proliferative index (R2 = 4 x 10(-8). CONCLUSION: The results of this study show no association between the nature of the intragenic NF2 gene mutation and tumor behavior. It is likely therefore that NF2 gene inactivation is not the only determinant of tumor behavior in vestibular schwannoma. PMID- 9391674 TI - Does the stapes reflex remain the same after Bell's palsy? AB - OBJECTIVE: The authors investigated the integrity and function of nervus stapedius 1 year after facial paralysis. STUDY DESIGN: Patients with Bell's palsy were observed prospectively for 1 year and compared with healthy patients. SETTING: The follow-up of patients was done in the outpatient clinic and tests were applied in the audiology unit. PATIENTS: The mean age of 32 patients was 41.03 years. Eight of 32 patients were grade II (25%), 11 were grade III (35%), and 13 were grade IV (40%) according to House-Brackman grading system. The mean age of the control group (10 persons) was 36.5 years. INTERVENTION: Contralateral stimulus was used in acoustic reflex test at 500 and 1,000 Hz with 80-, 90-, 100 , and 110-dB stimulus intensity. Tests were applied in three ways: normal position, eye-closed position, and grin position. Tests were done in the first 15 days of facial paralysis and repeated at least 1 year thereafter. The millimeter difference in amplitude of impedance recording of middle ear between the normal ear and paralyzed ear was accepted as criterion. MAIN OUTCOME MEASURES: There were 6- to 9-mm amplitude differences between normal side and healed side of grade IV patients with 100- and 110-dB stimuli. RESULTS: In the second test (after 1 year), statistically significant differences were present between control group and grade IV patients on 1,000 and 500 Hz frequencies with 100- and 110-dB stimulus intensity (p < 0.05). There were no significant differences between grade II and control group and between grade III and control group. CONCLUSIONS: A permanent partial denervation is present on the stapedial nerve, especially after grade IV paralysis, and it affects the function of stapes muscle in high decibel sounds. But it does not affect the stapes reflex threshold. No synkinetic innervation was found in the authors' patient group with their test method. PMID- 9391675 TI - Surface temperature distributions in carbon dioxide, argon, and KTP (Nd:YAG) laser ablated otic capsule and calvarial bone. AB - HYPOTHESIS: The spatial and temporal surface temperature distribution was measured after laser irradiation in fresh porcine otic capsule and calvarial bone tissue using an HgCdTe (mercury-cadmium-tellurium) infrared camera. BACKGROUND: Carbon dioxide (CO2) (lambda = 10.6 mm), argon (lambda = 514 nm), and Potassium Titanyl-Phosphate Neodynium: Yttrium-Aluminum-Garnet (KTP[Nd:YAG]) (lambda = 532 nm) lasers are used for stapes surgery and in the treatment of chronic ear disease. Despite extensive clinical use, little is known about the thermal perturbations in otic capsule calcified tissues and what are safe energy parameters for laser use. METHODS: A microspot manipulator, lens, and microfiber were used for continuous wave (CW) and super-pulse (SP) CO2, argon, and KTP(Nd:YAG) lasers, respectively. Peak temperatures after ablation were measured simultaneously along with the full-width--half-maximum of the thermal disturbance and fitted to a Gaussian distribution. The cooling time for the hot spot to return to ambient temperature also was recorded. RESULTS: Temperature changes with CW CO2 irradiation were markedly elevated relative to SP mode and also required longer to cool. The KTP and argon-treated bone were irradiated in the presence and absence of an initiator (black ink): minimal surface temperature elevation was recorded in the absence of an initiator. Further, no surface modification was observed. In contrast, the addition of an initiator resulted in marked temperature elevations and significant surface carbonization with these two visible wavelength lasers. Cooling times varied from 10-40 seconds. No consistent relation to the measured thermal values and tissue microarchitecture was observed. CONCLUSIONS: The measured cooling times and Gaussian distribution of surface temperatures serve as empiric guidelines for minimizing thermal injury to critical structures during laser surgery in the middle ear. PMID- 9391676 TI - Juvenile keratin inoculation induces chronic ear pathology. AB - OBJECTIVE: Human neonatal temporal bones frequently show the formation of granulation tissue provoked by amniotic fluid keratin contents, desquamated keratinized epithelial cells and lanugo hair. Similar histopathologic findings have been produced previously in a short-term animal model. To test the hypothesis that those short-term pathologic observations could have theoretical relevance for ear disease in older patients, a longer term animal model study was necessary. METHODS: Into the right bulla of 10 chinchilla pups was placed an aliquot of autogenous, nonviable epidermal scrapings and hair. Into the left bulla was placed 1 mm2 viable autogenous epidermal tissue. Animals were killed at intervals up to 11 months and then studied by light microscopy. RESULTS: Chronic ear histopathologic changes such as granulation tissue, osteoneogenesis, adhesions, and cholesteatoma were present. Over time, these secondary pathologic changes became more obvious than the initial keratin implant. CONCLUSIONS: The authors conclude that chronic pathologic changes resembling human ear disorders persist and that this model further extends the hypothesis that prenatally acquired keratin eventually could account for some cases of human ear disease. PMID- 9391677 TI - Imaging case of the month: the narrow internal auditory canal. PMID- 9391678 TI - Ear drop ototoxicity: reality or myth? PMID- 9391679 TI - Mechanism of anaphylactoid reactions: improper preparation of high-dose intravenous cyclosporine leads to bolus infusion of Cremophor EL and cyclosporine. AB - BACKGROUND: During a Phase I/II trial of high-dose intravenous cyclosporine, a high incidence of anaphylactoid reactions was observed. Epidemiologic investigations revealed that the occurrence of anaphylactoid reactions was significantly associated with improper mixing during preparation of the infusions. It was hypothesized that improper mixing during the preparation of the infusion may have caused initial bolus infusions of the vehicle, Cremophor EL. These inadvertent bolus infusions may have caused the anaphylactoid reactions. OBJECTIVE: To investigate the effect of different mixing techniques on the distribution of the components of cyclosporine concentrate for infusion: cyclosporine, Cremophor EL, and ethanol in the infusions administered to the patients. METHODS: Infusions were prepared in a similar fashion as those administered to study patients enrolled in a high-dose cyclosporine therapy protocol. Samples were collected at defined time points of the infusions. Concentrations of cyclosporine and Cremophor EL were spectrophotometrically determined; ethanol concentrations were measured enzymatically. RESULTS: Cyclosporine and Cremophor EL concentrations were up to ninefold higher than intended during the first 10 minutes of the infusions that were not appropriately mixed. In contrast, the concentrations of cyclosporine and Cremophor EL were similar to the intended concentrations in all of the well-mixed infusions. CONCLUSIONS: Inappropriate mixing of high-dose cyclosporine infusions can lead to initial bolus infusion of cyclosporine and Cremophor EL. Bolus infusions of Cremophor EL have been associated with anaphylactoid reactions. Thus, through mixing of high-dose cyclosporine infusions may be important to reduce the possibility of life-threatening anaphylactoid reactions. PMID- 9391680 TI - Formulation and stability of naltrexone oral liquid for rapid withdrawal from methadone. AB - OBJECTIVE: To assess the stability of naltrexone oral liquid prepared from tablets and powder, and to evaluate its use in precipitating rapid withdrawal from methadone. DESIGN: Naltrexone 1 mg/mL oral liquids were prepared from tablets and powder and stored in the dark at 4, 25, and 70 degrees C. Similar formulations containing 5 mg/mL were stored at 70 degrees C. The 1-mg/mL formulation prepared from tablets was clinically evaluated in inducing rapid withdrawal in two drug-dependent individuals receiving methadone maintenance treatment using a naltrexone dose titration protocol. SETTING: A university pharmacy school and affiliated urban teaching hospital. MAIN OUTCOME MEASURES: Samples removed at six time points were analyzed for naltrexone concentration to assess decomposition over 90 days. An opioid withdrawal symptom checklist was used to assess the severity of the withdrawal symptoms prior to, and 30 minutes after, each dose of naltrexone. RESULTS: Decomposition of naltrexone in all formulations stored at 4 and 25 degrees C was not significant over 90 days. Both patients tolerated naltrexone 1 mg/mL oral liquid, but found it bitter and gritty. Withdrawal symptoms were experienced immediately after the first dose, but were resolving by the end of day 3 of naltrexone treatment, at which stage both patients were able to tolerate a 50-mg tablet of naltrexone as maintenance. CONCLUSIONS: Naltrexone 1 mg/mL oral liquids prepared from tablets or powder are stable when stored in the dark for 60 days at 4 degrees C and for 30 days at 25 degrees C. The formulation prepared from tablets provides flexible dosing in patients undergoing rapid withdrawal from methadone. PMID- 9391681 TI - Drug samples and family practice residents. AB - OBJECTIVE: To describe residents' knowledge, attitudes, and behaviors regarding sample medications and to determine the education provided in residency training regarding sample use. METHODS: A 6-item survey was sent to directors of US family practice residency programs. Residents of a sample of these programs were sent an anonymous, self-administered, 21-item questionnaire assessing knowledge, attitudes, and practices relating to sample use. Both surveys consisted of initial and follow-up mailings. RESULTS: The residency directors' survey was returned by 232 of the 436 residency directors (53%). Although 66% of the programs had a policy regarding samples, only 15% of the policies completely incorporated recommendations of the Society of Teachers of Family Medicine. After two mailings, 248 resident responses were received from 43 of 47 residencies (92%). Only 21% of respondents thought that they received adequate training about sample use in medical school; this number increased to 49% for residency training. Agreement with the adequate training statement was highest among respondents from residencies that had both a sample distribution policy and a pharmacist (p = 0.044). Fifty-five percent thought that samples influenced their prescribing and 70% thought that samples helped them to learn more about the sampled medication. CONCLUSIONS: Family practice residents value and use samples, although they are often unaware of the rules governing the labeling of samples. While reported distribution of samples by residents often is appropriate, education about effective sample use could be improved. Drug samples play a significant role in residency training. PMID- 9391682 TI - Consumer requests for information regarding psychotropic drugs: experience from a national medicines phone-in. AB - OBJECTIVE: To undertake a qualitative analysis of calls regarding psychotropic drugs that were received during a national medicines phone-in day. BACKGROUND: In July 1996, The Society of Hospital Pharmacists of Australia coordinated a national medications phone-in day, allowing consumers to seek information about medications from pharmacists and physicians using a toll-free telephone number. METHODS: Data collection forms were used to record the details of all calls answered during the phone-in day. Demographic data collected included the estimated age and gender of the caller. Other data collected included the drugs that were the subject of the inquiry and the category of questions. RESULTS: There were 42,096 attempted connections to the service, but because of limited telecommunications capacity, only 2245 callers were successfully connected. Psychotropic drugs were the primary subject of 367 calls, representing 16.4% of all inquiries for which data collection forms were completed. Antidepressants (56.1%) and benzodiazepines (24.8%) were the two most commonly encountered classes of psychotropic drugs. The greatest proportion of calls (57.2%) was related to adverse effects of medications. The nature of the inquiries regarding adverse drug effects was generally consistent with the adverse effects detailed in the scientific literature. CONCLUSIONS: The results of this 1-day, consumer oriented drug information project suggest that there is a substantial need for this type of service. Patients treated with psychotropic medications should have access to unbiased, high-quality information about drug therapy. PMID- 9391683 TI - Continuous intrathecal meperidine via an implantable infusion pump for chronic, nonmalignant pain. AB - OBJECTIVE: To report a continuous infusion of intrathecal meperidine via an implanted infusion pump for nonmalignant, chronic pain. CASE SUMMARY: A 69-year old white woman had chronic, nonmalignant low-back pain and bilateral leg pain. Multiple drug therapies and other interventional techniques had failed. The patient achieved significant pain relief by a continuous infusion of intrathecal meperidine via an implanted infusion pump. DISCUSSION: To our knowledge, this is the first report of meperidine administered intrathecally by continuous infusion. Continuous infusion of intrathecal and epidural opiates by implanted infusion pumps is becoming more widely recognized as an alternative treatment for patients with chronic, benign pain. Epidural and intrathecal meperidine is an effective analgesic for short-term surgical procedures. Data reporting effective relief and safety with continuous intrathecal meperidine remain limited. CONCLUSIONS: Continuous intrathecal meperidine via an implantable infusion pump may be an effective alternative in the treatment of chronic pain. PMID- 9391684 TI - Symptomatic syndrome of inappropriate antidiuretic hormone secretion associated with azithromycin. AB - OBJECTIVE: To report a case of symptomatic syndrome of inappropriate antidiuretic hormone (SIADH) secretion associated with azithromycin and review the literature related to this adverse drug reaction. DATA SOURCES: Review articles identified by a computerized (MEDLINE) (1966-April 1996) and manual (Index Medicus) search. DATA SYNTHESIS: Azithromycin is a well-tolerated broad-spectrum macrolide antibiotic. We report a symptomatic case of SIADH secretion associated with azithromycin. The patient received two doses of azithromycin before the development of sudden mental status changes associated with severe hyponatremia. All other potential causes were ruled out. No previous reports exist in the literature. CONCLUSIONS: Azithromycin may be associated with symptomatic SIADH secretion. Awareness and attention are required if patients develop mental status changes or hyponatremia while receiving azithromycin so that appropriate diagnostic and therapeutic actions can be implemented. PMID- 9391685 TI - Use of estrogen therapy in a patient with gastrointestinal bleeding secondary to arteriovenous malformations. AB - OBJECTIVE: To describe a patient with gastrointestinal (GI) bleeding caused by arteriovenous malformations (AVMs) that was treated with estrogen therapy. CASE SUMMARY: A 70-year-old white man was diagnosed with multiple AVMs in the cecum, duodenum, and stomach. Pharmacologic management included the use of ferrous sulfate; however, the patient continued to have recurrent bleeding that required multiple transfusions and endoscopic cauterization. Therapy was initiated with ethinyl estradiol 0.05 mg po qd; no further transfusions have been required for 10 months. DISCUSSION: It is estimated that AVMs of the GI tract account for 1-8% of upper GI bleeding episodes and up to 6% of lower GI bleeding episodes. Hormonal agents have been reported to decrease bleeding in patients with both hereditary and acquired AVMs. CONCLUSIONS: The role of estrogen therapy in treating AVMs of the GI tract is unclear and supported by only one clinical study. PMID- 9391686 TI - Baclofen toxicity in patients with severely impaired renal function. AB - OBJECTIVE: To report the toxic effects of baclofen in patients with severely impaired renal function. DATA SOURCES: From 1991 to 1995, nine patients with severely impaired renal function (2 not receiving dialysis, 1 undergoing continuous ambulatory peritoneal dialysis [CAPD], and 6 receiving maintenance hemodialysis), who exhibited clinical toxicity after baclofen therapy at our hospital were included for analysis. Another seven cases from the literature obtained by computerized (MEDLINE) and manual (Index Medicus) search methods published between 1980 and 1995 were also reviewed. INTERVENTION: Among our nine patients, the six undergoing chronic hemodialysis and one not undergoing dialysis received early (< 48 h) hemodialysis after toxic symptoms developed. The patient undergoing CAPD received late hemodialysis (> 72 h), and the other patient who had not undergone dialysis received only supportive care. RESULTS: A review of these 16 cases revealed that most patients received only small doses and very short-term baclofen therapy. Altered consciousness was the major presenting feature. Severe acute complications, such as seizures and respiratory depression, were relatively uncommon among patients with severely impaired renal function. However, abdominal pain, which has previously rarely been reported, was noted in five of our nine patients. Most patients showed clinical improvement after hemodialysis. An analysis of these nine patients revealed that those who received early hemodialysis had a shorter recovery time than the patient who received only supportive care (2.71 +/- 0.42, respectively, vs. 9 d; p < 0.01). A lag of several hours between the end of the hemodialysis session and an improvement in the level of consciousness was noted. DISCUSSION: As most patients with severely impaired renal function developed toxic symptoms soon after initiating a low-dose baclofen regimen, the accumulated dosage was small and severe complications were less common. Abdominal pain may have occurred as a result of the gamma aminobutyric acid-mediated cholinergic effect exerted by baclofen. The delay in conscious recovery after hemodialysis may be due to a delay in the clearance of baclofen from the central nervous system. CONCLUSIONS: Patients with severely impaired renal function generally develop baclofen intoxication soon after the initiation of low-dose therapy. Thus, the administration of baclofen, regardless of the dosage, in these patients is not appropriate. Abdominal pain, in addition to altered consciousness, is a common presenting feature in patients with renal failure who have baclofen intoxication. Hemodialysis is effective in alleviating the clinical symptoms and shortening the recovery time for such patients. PMID- 9391687 TI - Agranulocytosis induced by vancomycin or ticarcillin/clavulanate. AB - OBJECTIVE: To reacquaint clinicians with a reportedly rare adverse event of agranulocytosis occurring after long-term administration of vancomycin and ticarcillin/clavulanate, with a subsequent review of other reported cases in the literature. CASE SUMMARY: A 45-year-old white woman with spina bifida developed agranulocytosis (2.7 x 10(3)/mm3 white blood cells with only 3% polymorphonuclear leukocytes and no reported eosinophils or basophils) after long-term administration of vancomycin and ticarcillin/clavulanate for decubitus ulcers and chronic osteomyelitis. Consequently, the cell counts rebounded rapidly on discontinuation of both medications and returned to normal within 1 week. DISCUSSION: The incidence of vancomycin-associated neutropenia is presumably rare, but the increased use of vancomycin may disclose a more frequent occurrence. It is suggested that the mechanism for the reaction is immunologically mediated, yet this remains unclear. Although it is difficult to determine the causative agent in this case, vancomycin was most suspect clinically. Ticarcillin/clavulanate is less likely because our patient has since been readmitted and treated with oxacillin, imipenem/cilastatin, and amoxicillin/clavulanate without affecting the white blood cell count. In that regard, it could be reasoned that an immunologic reaction to ticarcillin would have resulted in a similar outcome with other penicillins. CONCLUSIONS: This case serves as a reminder to clinicians that patients receiving long-term treatment with vancomycin should have their white blood cell count monitored at least weekly. PMID- 9391688 TI - Olanzapine: a new antipsychotic agent with efficacy in the management of schizophrenia. AB - OBJECTIVE: To review the pharmacology, pharmacokinetics, efficacy data, and adverse effects of olanzapine as a treatment for schizophrenia and to determine the advantages and disadvantages of this atypical antipsychotic agent compared with currently marketed agents. DATA SOURCES: A MEDLINE computer literature search was conducted to retrieve all English-language studies and review articles involving olanzapine published as of October 1, 1996. The manufacturer of the drug, Eli Lilly and Company, provided the clinical investigator's brochure and abstracts of unpublished Phase III clinical trials. STUDY SELECTION: Animal studies evaluating the pharmacology of olanzapine were evaluated, as were all open-label and double-blind studies involving the evaluation of olanzapine for the treatment of patients with schizophrenia. DATA EXTRACTION: All available clinical studies were reviewed and the interpretation of data for each study was influenced by the size of the study sample, the nature of the inclusion and exclusion criteria, and the data analysis techniques used. DATA SYNTHESIS: Olanzapine is a thienobenzodiazepine analog with an in vitro receptor affinity profile similar to that of clozapine. Olanzapine exhibits linear kinetics over the dosage range studied and is extensively metabolized in humans. Clinical evaluations to date have shown olanzapine to be at least as efficacious as typical antipsychotic agents in the treatment of the acute phase of schizophrenia. The drug was well tolerated, with significantly fewer extrapyramidal adverse effects than haloperidol. Current data suggest that olanzapine may be more effective than haloperidol for the treatment of negative symptoms; moreover, preliminary data suggest that fewer relapses occur over the course of treatment in patients treated with olanzapine compared with those taking haloperidol. CONCLUSIONS: The exact place of olanzapine in the therapy of psychotic patients remains unclear, as more data are needed to evaluate the long term efficacy of this agent, its impact on negative symptoms, and its potential use in patients resistant to the standard agents. Despite limitations in the current database, olanzapine is a promising treatment option for patients with schizophrenia. PMID- 9391689 TI - Recombinant human tumor necrosis factor receptor (p75) Fc fusion protein (TNFR:Fc) in rheumatoid arthritis. AB - BACKGROUND: Tumor necrosis factor (TNF) is the dominant mediator of the cytokine cascade that causes inflammation and joint destruction in rheumatoid arthritis. A new class of agents under investigation, the biologic TNF inhibitors, inhibits the activity of TNF. Recombinant human TNF receptor p75 Fc fusion protein (TNFR:Fc; Enbrel) blocks the activity of the cytokine TNF. The preclinical, Phase I, and Phase II data of TNFR:Fc in rheumatoid arthritis are reviewed in this article. METHODS: All available data on TNFR:Fc in rheumatoid arthritis were reviewed. These data included published literature and data on file at the manufacturer. RESULTS: TNFR:Fc has been effective in many models of inflammation, including animal models of rheumatoid arthritis and in clinical rheumatoid arthritis trials. Conclusions from a study with TNFR "knockout" mice (genetically altered mice incapable of producing TNFR proteins) demonstrated that p75 TNFR is a natural antagonist of TNF-mediated inflammation. A placebo-controlled, dose escalation, Phase I trial evaluated the safety and efficacy of TNFR:Fc in patients with rheumatoid arthritis. There were no serious adverse effects reported. A Phase II, randomized, double-blind, placebo-controlled trial evaluated 180 patients with active rheumatoid arthritis whose previous therapy had failed. A dose-response relationship was observed in the number of tender and swollen joints; patients who received the highest dose (16 mg/m2) of TNFR:Fc had the greatest improvement. Treatment was generally well tolerated. TNFR:Fc is nonimmunogenic; no antibodies to TNFR:Fc have been detected thus far in human studies. CONCLUSIONS: Preliminary data indicate that TNFR:Fc is an excellent candidate for future long-term studies in the treatment of rheumatoid arthritis. PMID- 9391690 TI - Prehospital-initiated thrombolysis. AB - OBJECTIVE: To evaluate the feasibility, safety, and efficacy of prehospital initiated thrombolysis in decreasing the mortality rate due to acute myocardial infarction. DATA SOURCES: English-language clinical studies, abstracts, and review articles identified from MEDLINE searches and bibliographies of identified articles. Epidemiologic data were extracted from the Internet. STUDY SELECTION: Eight randomized clinical trials and two meta-analyses that compared prehospital initiated thrombolysis with in-hospital-initiated thrombolysis. DATA EXTRACTION: Pertinent studies were selected and the data were synthesized into a review format. DATA SYNTHESIS: Early reperfusion of an infarct-related coronary artery is associated with lower mortality rates. Most of the delay in initiating treatment is caused by patient delay rather than transport delay or hospital delay. In addition, more than 30% of eligible patients do not receive thrombolytic therapy. Prehospital initiation of thrombolysis has been evaluated as a means of decreasing hospital delay and increasing the number of eligible patients receiving thrombolysis. Clinical trials document that prehospital initiated thrombolysis is feasible and safe, and saves time. Of the eight randomized trials, three demonstrated a decrease in either cardiac or total mortality with prehospital thrombolysis. All studies were limited by relatively small sample sizes. Two published meta-analyses suggest a 16-17% reduction in mortality with prehospital thrombolysis. In the US, prehospital thrombolysis is not routinely recommended due to medical issues related to diagnostic accuracy and monitoring, legal concerns, and economic implications. Additional strategies, such as community-wide education and prehospital diagnostic electrocardiograms (ECGs), are being studied. CONCLUSIONS: In clinical trials, prehospital-initiated thrombolytic therapy was shown to be safe and probably more effective than in hospital administration of thrombolytic therapy, but this has not proven feasible in the US at this time. Despite time-savings by decreasing treatment delay with prehospital-initiated thrombolysis, patient delay still persists and accounts for the majority of delay. Future investigations will center on increasing the number of patients treated with thrombolytic agents through patient education, in patient and out-patient programs that rapidly identify eligible patients, as well as prehospital diagnostic ECGs. PMID- 9391691 TI - Pharmacologic management of supraventricular tachycardias in children. Part 2: Atrial flutter, atrial fibrillation, and junctional and atrial ectopic tachycardia. AB - OBJECTIVE: To review the literature regarding the use of antiarrhythmic agents in the management of atrial flutter (AF), atrial fibrillation (Afib), junctional ectopic tachycardia (JET), and atrial ectopic tachycardia (AET) in infants and children. To discuss the advantages and disadvantages of specific agents in each type of arrhythmia in an effort to develop treatment guidelines. DATA SOURCE: A MEDLINE search encompassing the years 1966-1996 was used to identify pertinent literature for discussion. Additional references were found in the articles, which were retrieved via MEDLINE. STUDY SELECTION: Clinical trials that address the use of antiarrhythmic agents for the treatment of supraventricular tachycardia, AF, Afib, JET, and AET in children were selected. Literature pertaining to dosage, pharmacokinetics, efficacy, and toxicity of antiarrhythmic agents in children were considered for possible inclusion in the review; information judged to be pertinent by the authors was included in the discussion. DATA EXTRACTION: Although there are numerous reports of antiarrhythmic use in children, there are very few large studies designed that evaluate the use of specific antiarrhythmic agents in the treatment of AF, Afib, JET, or AET. Ideally, controlled clinical trials are used to develop clinical guidelines; however, in this situation, most data and information must be obtained from case series of children treated. Although the results from these types of studies may be useful in developing guidelines for the optimal use of these agents for the treatment of AF, Afib, JET, and AET, controlled trials are required for establishing standard treatment guidelines for all patients. DATA SYNTHESIS: Despite limited scientific evaluation of conventional agents in the treatment of AF, Afib, JET, or AET in children, they continue to be the standards of care. Most information regarding the use of conventional agents in children has been extrapolated from the adult literature. Little justification for the use of the agents or dosing in children is available. Controlled trials regarding the use of newer antiarrhythmic agents (propafenone, amiodarone, flecainide) are available; however, the variance in dosing schemes, presence of structural heart disease, and patient age may confound the results. CONCLUSIONS: Because of greater clinical experience, conventional antiarrhythmic agents generally remain as first line therapy in the management of most supraventricular tachycardias in children. Atrial pacing or cardioversion to reestablish sinus rhythm is indicated for initial episodes of AF in infants, followed by chronic prophylactic therapy in those with significant structural heart disease or in infants in whom AF recurs. Attempts to eliminate AF in children outside the neonatal or infancy period should begin with trials of traditional agents such as digoxin or procainamide, and if unsuccessful, subsequent trials of amiodarone. Digoxin and beta-blockers remain the mainstay of therapy for children with Afib, followed by procainamide for treatment failures. Intravenous amiodarone, the newest addition to our antiarrhythmic armamentarium, is the most promising agent in the treatment of postoperative JET. This arrhythmia has been traditionally managed with corporal cooling and/or digoxin therapy; however, intravenous amiodarone may now be a valuable option. Although relatively unsuccessful in the management of congenital JET and AET, conventional agents are typically used prior to the initiation of long-term therapy with potentially more toxic agents such as amiodarone or propafenone. Additional well-designed, controlled trials are needed to further evaluate the comparative efficacy of agents such as flecainide, sotalol, moricizine, propafenone, and amiodarone in the management of AF, Afib, JET, and AET in children, as well as to evaluate the dosing and toxicity in various age groups. PMID- 9391692 TI - Ethnicity and antipsychotic response. AB - OBJECTIVE: To review the data generated by studies examining interethnic/racial differences in response to antipsychotics. DATA SOURCES: A MEDLINE search (1966 1996) identified all articles examining differences in antipsychotic response among Caucasians, Asians, Hispanics, and African-Americans, as well as articles evaluating postulated mechanisms for these differences. STUDY SELECTION: All abstracts, studies, and review articles were evaluated. DATA SYNTHESIS: Ethnic/racial differences in response to antipsychotic medications have been reported and may be due to genetics, kinetic variations, dietary or environmental factors, or variations in the prescribing practices of clinicians. Studies suggest that Asians may respond to lower doses of antipsychotics due to pharmacokinetic and pharmacodynamic differences. Research relevant to African Americans is limited, but some studies suggest that differences in this group may be due to clinician biases and prescribing practices, rather than to pharmacokinetic or pharmacodynamic variability. CONCLUSIONS: Future research directed at validating the hypotheses that different ethnic/racial groups show variations in response to antipsychotics should focus on homogeneous ethnic groups, use recent advances in pharmacogenetic testing, and control for such variables as observer bias, gender, disease chronicity, dietary and environmental factors, and exposure to enzyme-inducing and -inhibiting agents. Clinicians should be aware that potential interethnic/racial differences in pharmacodynamics and pharmacokinetics may exist that can alter response to antipsychotics. PMID- 9391693 TI - Muromonab-CD3 and antithymocyte globulin in renal transplantation. AB - OBJECTIVE: To review the recently published medical literature for the practical and efficient use of muromonab-CD3 (OKT-3) and antithymocyte globulin (ATG) in renal transplantation. DATA SOURCES: MEDLINE and EMBASE were searched (1985 February 1996). Key words used were antithymocyte globulin (ATG, Atgam), muromonab-CD3 (OKT-3, Orthoclone), and kidney transplantation. Thereafter, the search was restricted to English-language articles, clinical trials, and human studies. STUDY SELECTION AND DATA EXTRACTION: The search was reviewed for articles of interest, and pertinent references from these articles were further reviewed to supplement the initial search. The review focused on antibody therapy as induction and/or rejection therapy in renal transplantation. DATA SYNTHESIS: Although ATG and OKT-3 are effective in delaying and reducing the occurrence of acute rejection, their impact on long-term graft survival has not been established. Improved graft survival has, however, been demonstrated in patients at high risk for rejection. These risks are described in the review. As first line or steroid-resistant rejection therapy, ATG and OKT-3 have proven efficacious. Some studies have shown improved graft survival with OKT-3. Although serious infections may occur, OKT-3 has been shown to be effective in reversing rejections resistant to both steroids and ATG. Therefore, reserving OKT-3 for steroid- or ATG-resistant rejections may be preferred over the first-line use of OKT-3, which is limited by the development of antimurine antibodies with subsequent uses. However, the benefits of first-line antibody therapy may outweigh the risks of developing these antibodies in patients for whom high-dose steroids may not be the most appropriate treatment. Other factors that need to be considered are adverse effects, which appear to be lower with ATG, cost, and total hospital charges. The accuracy of treatment outcomes analysis among these studies is limited by variations in the immunosuppressive regimens of the study centers, doses of concomitant therapies, use of prophylactic antibiotics, and time to follow-up. CONCLUSIONS: While important benefits are realized from using antibody therapies in renal transplantation, their use is often associated with excess immunosuppression and increased treatment costs. Despite encouraging results from published trials, questions regarding the extent of their prophylactic use and impact on long-term outcomes need to be answered. The current literature contains no prospective, controlled, randomized comparisons of OKT-3 and ATG with standardized regimens of conventional immunosuppressive agents and antirejection protocols. The majority of studies use OKT-3 as part of the treatment protocol. Well-designed studies using ATG are lacking. Further research is needed to refine treatment protocols for ATG and OKT-3 to determine the optimal timing and dosing for these agents. PMID- 9391694 TI - The role of reactive drug metabolites in immune-mediated adverse drug reactions. AB - OBJECTIVE: To highlight recent advances in the understanding of adverse drug reactions (ADRs), with a focus on models outlining interactions between drug metabolism, disease processes, and immunity. Specific mechanisms that identify the metabolic pathways responsible for drug bioactivation to reactive drug metabolites (RDMs) involved in the initiation and propagation of specific immune mediated hypersensitivity reactions are discussed. Drug classes well known to be associated with immune-mediated ADRs are reviewed and the clinical implications of current research are discussed. DATA SOURCES: Original experimental research and immunologic review articles relevant to ADR diagnosis and etiology. DATA EXTRACTION: Results of relevant in vitro experiments and clinical reactions to drug therapy were compiled and reviewed. Critical discoveries concerning the identification of RDMs involved in ADRs were highlighted, with respect to RDM involvement in the production of an immune response to drug haptens. DATA SYNTHESIS: Drug adverse effects are classified according to clinical characteristics, immune interactions, and mechanistic similarities. Cytochrome P450 bioactivation of drug molecules to RDMs is a prerequisite to many ADRs. An electrophilic metabolite may react with cellular macromolecules (i.e., lipids, proteins, nucleic acids), resulting in direct cellular damage and organ toxicity. Covalent binding of an RDM to cellular macromolecules may also result in the formation of a hapten that is capable of eliciting a cellular or humoral immune response against drug or protein epitopes, culminating in the characteristic symptoms of hypersensitivity reactions. Mechanistic details concerning the identification of stable protein-metabolite conjugates and their interaction with the immune system remain unclear. Genetic imbalance between bioactivation and detoxification pathways, as well as reduced cellular defense against RDMs due to disease or concomitant drug therapy, act as risk factors to the onset and severity of ADRs. CONCLUSIONS: Adverse reactions to drug therapy cause significant morbidity and mortality. Identification of the pathways involved in drug bioactivation and detoxification may elucidate the potential of chemical agents to induce immune-mediated ADRs. Understanding the mechanisms of ADRs to current xenobiotics is helpful in the prevention and management of ADRs, and may prove useful in the design of novel therapeutic agents with reduced incidence of severe adverse events. PMID- 9391695 TI - Losartan versus ACE inhibitors in the treatment of hypertension. PMID- 9391696 TI - Vaginal misoprostol for term labor induction. AB - Misoprostol is an effective agent for cervical ripening and induction of labor. The use of oxytocin was significantly decreased in patients treated with misoprostol versus dinoprostone. It has been used to induce over 1000 women in reported studies and has demonstrated a safety profile comparable with that of endocervical and vaginal dinoprostone. Uterine hyperstimulation was a concern in earlier trials, but at a reduced dose of 25 micrograms, the incidence has decreased to a level that is comparable with the values reported for dinoprostone. Misoprostol tablets are stable at room temperature and are considerably less expensive than the dinoprostone alternatives. Two additional factors pertaining to misoprostol administration must be taken into account before the drug is selected for vaginal use. First, Cytotec tablets are currently available in two strengths, 100 and 200 micrograms. This can lead to confusion or error if the clinician orders a quarter or half tablet. The order should always identify the strength in micrograms (25 or 50 micrograms). Second, the 100 microgram tablet is not scored; therefore, the proper dose should be carefully prepared by a pharmacist using a pill cutter. Key members of the hospital staff must be trained about the proper use of misoprostol for labor induction before initiating therapy. One alternative to directly inserting the tablet is to pulverize it and mix with a gel such as hydroxyethylcellulose gel. However, such compounding introduces the same problems with stability and uniformity of dose as experienced with dinoprostone gels. Despite the success of misoprostol in clinical trials, it is not approved for this indication, and the manufacturer of Cytotec does not plan to pursue approval. Therefore, independent, large-scale studies are warranted to more accurately assess the efficacy and overall safety of using intravaginal alprostadil for cervical ripening and labor induction. Additional clinical experience should also help to determine the best regimen and method of administration. From the data currently available, it appears that either a 25- or 50-microgram dose (one-fourth or one-half of a 100-microgram tablet) inserted into the posterior vaginal fornix and repeated at 4-5-hour intervals if needed, is a clinically effective regimen, and is associated with the least amount of adverse effects and complications. As with all labor inductions, uterine contractions and fetal heart rate should be monitored carefully throughout the procedure. PMID- 9391697 TI - Hydroxyurea in the treatment of sickle-cell anemia. AB - Sickle-cell anemia is a congenital hemolytic anemia characterized by sickle shaped RBCs. The deformed RBCs become distorted and rigid and may occlude small arterioles and capillaries leading to tissue ischemia and infarction. Sickled RBCs are too fragile to withstand the trauma of circulation, and hemolysis occurs after they enter the circulation. RBCs with a high level of Hb F are resistant to sickling. Hydroxyurea has been shown to stimulate Hb F synthesis, leading to a reduction in the incidence of hemolytic and vaso-occlusive manifestations; however, hydroxyurea has no role in the treatment of crises already in progress. The National Heart, Lung, and Blood Institute announced in January 1995 that treatment with hydroxyurea leads to an increase in Hb F production within RBCs and a reduction in the frequency of painful crises in patients with sickle-cell anemia. Although the mechanism by which hydroxyurea increases Hb F is not known, one possible explanation is that hydroxyurea is cytotoxic to the more rapidly dividing late erythroid precursors, leading to the recruitment of early erythroid precursors that have demonstrated increased capacities to produce Hb F. Clinical trials have demonstrated that hydroxyurea results in an increase in Hb F concentrations; however, this increase may not dramatically affect the progressive vascular changes associated with sickle-cell anemia; thus, patients may still experience complications related to sickle-cell anemia. At North Carolina Baptist Hospital in Winston-Salem, NC, compliant patients with sickle cell anemia are started on hydroxyurea. There are no specific criteria for patient selection or monitoring. The dosage is started at 10-15 mg/kg/d. Platelet count, complete blood count, and Hb F are monitored and hydroxyurea dosages are adjusted accordingly. Although hydroxyurea has been effective in the treatment of sickle-cell anemia, large double-blind, placebo-controlled clinical trials are needed to determine whether the risks of long-term administration outweight the risk of vaso-occlusive disease in untreated patients. PMID- 9391698 TI - Perspectives on alternative medicine. PMID- 9391699 TI - Induction of labor: a clinician's viewpoint. PMID- 9391700 TI - Methods for preventing reactions secondary to Cremophor EL. PMID- 9391701 TI - Rifabutin-associated uveitis. PMID- 9391702 TI - Absence of cross-reaction between lisinopril and enalapril in drug-induced lupus. PMID- 9391703 TI - Hypertension exacerbated by amphotericin B administration. PMID- 9391704 TI - Psychotic episode after melatonin. PMID- 9391705 TI - Comment: zolpidem: distinct from triazolam? PMID- 9391706 TI - Correction and comment: possible toxicity from propylene glycol in injectable drug preparations. PMID- 9391707 TI - Prenatal fluoride for growth and development: Part X. AB - Examinations of prenatal fluoride supplemented (PNF) teeth in an animal model and in a five-month human fetus find these teeth to be more developed than the non supplemented controls. The fact that PNF allows teeth to develop to their full potential suggests that PNF could be an essential nutrient for the entire human and this could be demonstrated most easily during rapid fetal growth. A review of the recent literature, including trials by NIH and The World Health Organization, provide evidence that fluoride (F) does allow the fetus to grow and develop to its full potential. The authors conclude that PNF must be supplied in at least a 2 mg/day pulse dose, and then F must be given from shortly after birth in a daily amount appropriate for the weight of the child with some consideration for the amount of F water utilized. PMID- 9391708 TI - Linguistic maturity as a determinant of child patient behavior in the dental office. AB - Progressively during the 20th century dentistry for children has become more efficient, less painful, and more prevention oriented. In the last quarter of the 20th century there was a dramatic decrease in dental decay for many American children. These two facts paired with the fact that stories about dentistry being painful are gone in many American communities and have been replaced with stories about how pleasant the dental appointment can be would seem to predict that child patient management and the interception of inappropriate behavior would not be a critical skill for the dental clinician that treats children today. This finding however is not the case. It is submitted that misbehavior now stems from the fact that today's parents are not encouraged to raise their children as urgently as in the earlier part of the century. It is offered that the child's incompetence in working with other people in the constituent speech acts of requests and promises causes the child confusion, frustration, and perhaps anxiety. The child's dental experience is a complex conversation between the dentists as requester and the child patient as the promisor of effective actions to the dentists' reasonable requests. PMID- 9391709 TI - The pulp capping procedure in primary teeth "revisited". AB - The purpose of this review is to "revisit" an earlier paper (1992) on the subject of direct pulp capping in primary teeth and bring some new considerations for the procedure by the use of dentin bonding adhesives. It has come to be recognized that the customary employment of calcium hydroxide for this therapy has some shortcomings that reduce the prognosis for a favorable outcome. For at least a decade, many investigations have found that postoperative sensitivity, thermal stimuli, pulp inflammation and pathosis can be attributed not to the composition of various dental materials and their insertion techniques, but to microleakage with subsequent bacterial invasion at the enamel/restoration and the dentin/pulp interfaces. It is imperative, as pointed out, that there be an impervious resinous bond between the dentin and the dentinopulpal complex which can be achieved by the use of dentinal adhesive agents to eliminate microleakage outward movement of pulpal fluids. Various steps in the bonding technique for the treatment of deep dentin caries and/or a pulp exposure has raised some concerns for their effect on the pulp. This review discusses these concerns, which can lead to the conclusion that the use of dentinal bonding adhesives is a safe and biologically feasible procedure, whether it be in permanent or primary teeth. PMID- 9391710 TI - Comparison of two tooth-saving preparation techniques in a treatment approach of one-surface cavities: design of a study. PMID- 9391711 TI - Uprighting the mandibular molars stimulates mandibular growth during treatment of class II malocclusion. AB - We hypothesized that uprighting of the mandibular molars creates a counter clockwise rotation of the mandible and stimulates mandibular forward growth during the treatment of a Class II malocclusion. This investigation used 33 longitudinal lateral cephalometric radiographs of Class II, Division 1 female patients. All cases were treated with non-extraction. Treatment was started in early adolescence with .018 slot edgewise Alexander appliances. High-pull head gear and Class II elastics were used. Seventeen cases that showed more than 5 degrees of uprighting of the mandibular first molars were selected as the uprighted group. Cases that showed less than 5 degrees of uprighting of the mandibular first molars were selected as the non-uprighted group. There was a significant correlation coefficient between the uprighted degree of the mandibular first molars and the degree of clockwise rotation of the mandibular plane to FH. PMID- 9391712 TI - Effect of acid-etching on fluoride-treated caries-like lesions of enamel: a SEM study. AB - Etching patterns by 20 percent phosphoric acid on caries-like lesions of enamel (white spot) untreated and treated with 0.4 APF were studied at 30, 60, 120 seconds of etch-times using the Scanning Electron Microscope (SEM). The surface topography of acid-etched teeth varied according to the etch time. Acid etching of caries-like lesions treated with fluoride showed etching patterns similar to sound enamel. Based on Silverstone's classification, thirty seconds etch-time produced type III pattern of surface morphology, while type I and type II were observed with 60 and 120 seconds of etch time. Fluoride treated lesions showed increased porosity and in addition to a surface coating of numerous small globules of calcium fluoride. Low level of topical fluoride before sealant application should be beneficial, since this allows a more rapid rate and increased degree of remineralization and possible arrest in the progress of caries lesions. PMID- 9391713 TI - Children of divorce. AB - Limited attention has been directed in the dental literature to the emotional, economic and associated consequences of divorces on children. A general introduction is provided on 1) the numbers of children involved in divorces in different single-parent population groups, with 2) emphasis on the emotional impact of divorce on children and 3) the potential significance for pediatric dental practices. PMID- 9391714 TI - Minority children in single-parent families. AB - Information is now available from the Bureau of the Census with details about the increasing number of single-parent families. A summary review is provided with particular emphasis on minority families. PMID- 9391715 TI - Caries prevalence in Ashkelon children in 1994. AB - In the five-year-old group, 182 children were examined, ninety males and ninety two females. Forty-three percent of the children were found to be caries-free with a dmf(t) of 2.08 + 2.64 (mean +/- S.D.). More boys than girls were caries free (46 percent vs 41 percent). In general, caries prevalence is lower and dental health is better in boys than girls. The f(t) component is 0. Children at this age are not treated. In the twelve-year-olds group, 129 boys and 132 girls (total of 261) were examined. Forty-one percent of the children were found to be caries-free with a DMF(T) of 1.43 + 1.70. In this group more girls than boys were caries-free (43 percent vs 39 percent). In general caries prevalence is higher, treatment levels are lower in boys than in girls in this age-group. The results show that dental health is better in Ashkelon children than in other partly fluoridated areas in Israel. Dental treatment levels are higher and caries prevalence is lower in Ashkelon than in comparable places in the country. The WHO goals for 2000 were achieved in Ashkelon by 1994. PMID- 9391716 TI - Ectodermal dysplasia with associated double tooth. AB - The case describes a double molar tooth in a seven-year-old girl who has ectodermal dysplasia. The most characteristic dental findings in ectodermal dysplasia are hypodontia and conically shaped crowns. In our case a double tooth was also present in the primary molar region, in addition to these characteristic findings. PMID- 9391717 TI - Veno-occlusive disease of the liver after bone marrow transplantation: is hypercoagulability really part of the problem? AB - Patients undergoing bone marrow transplantation (BMT) experience changes in various proteins with important functions in the coagulation and fibrinolysis system. Veno-occlusive disease (VOD) of the liver is a leading cause of non relapse mortality after BMT. Because of the concurrent occurrence of changes in the coagulation and fibrinolysis system and development of VOD, most authors assume a causative relationship between these two observations, but the results leading to this conclusion are not unequivocal. Data currently available do not allow the conclusion that coagulation activation and local excess fibrin generation are key factors in the pathogenesis of VOD. One approach to deciding whether there is, in fact, excessive local fibrin generation during the development of VOD might be the monitoring of high risk patients with tools that enable differentiation of local and systemic hypercoagulability (e.g. anti-D dimer immunoscintigraphy). Screening of patients at risk for VOD with special coagulation parameters pretransplant does not seem appropriate at present. However, markedly increased plasminogen activator inhibitor-1 levels (from baseline) seem to be appropriate tool to confirm the diagnosis of VOD when clinical suspicion exists. More research is needed in order to advance our understanding of the disease and to improve outcomes in both the prophylaxis and treatment of VOD. PMID- 9391718 TI - Increased soluble P-selectin following myocardial infarction: a new marker for the progression of atherosclerosis. AB - Increased soluble P-selectin has been described in atherosclerosis, but the mechanisms for this and its clinical significance are unknown. In an attempt to clarify these points we measured soluble P-selectin and von Willebrand factor, an endothelial cell marker, by ELISA in 116 patients who had survived a myocardial infarction and in 116 matched controls. Raised levels of both soluble P-selectin (median 272 ng/ml, range 55-850 ng/ml vs 190 ng/ml, range 40-395 ng/ml) and von Willebrand factor (mean +/- SD 128 +/- 37 IU/dl vs 100 +/- 33 IU/dl; both P < 0.001) failed to correlate (r = 0.12), and soluble P-selectin failed to correlate with any of the major risk factors for atherosclerosis. A four-year follow-up of 68 of these patients revealed that soluble P-selectin was higher in the 33 (48%) who had suffered an additional cardiovascular event (e.g. subsequent myocardial infarction, arterial surgery; median 350 ng/ml, range 275-460 ng/ml) compared with those free of an end-point (270 ng/ml, range 140-400 ng/ml, P = 0.0012). We conclude that increased soluble P-selectin is unrelated to the risk factors for atherosclerosis but is a new marker of disease progression in patients who have survived a myocardial infarction. PMID- 9391719 TI - Plasma levels of thrombomodulin and lipoprotein (a) in patients with cerebral thrombosis. AB - To evaluate the clinical implications of soluble thrombomodulin and lipoprotein (a) [Lp(a)] in patients with cerebral thrombosis, these parameters were measured in the plasma of 28 patients with cerebral thrombosis within 3 days of onset, 36 with cerebral thrombosis more than 1 month after onset, six with cerebral hemorrhage more than 3 months after onset and 37 healthy volunteers. In the patients with chronic-phase cerebral thrombosis, the thrombomodulin and Lp(a) levels were significantly higher and the total cholesterol level was significantly lower than in the normal group, while the patients with acute-phase cerebral thrombosis had significantly lower total cholesterol levels. The plasma level of Lp(a) in acute-phase cerebral thrombosis, but not that of thrombomodulin, was significantly higher in thromboses located in the cortex area and in patients with recurrent attacks than in the normal controls. There were no significant differences in thrombomodulin, Lp(a) or total cholesterol levels between the chronic-phase cerebral hemorrhage and normal groups. These findings support the hypothesis that Lp(a) plays a part as a risk factor in cerebral thrombosis, especially in patients with a cortex area thrombosis and in patients with a recurrent attack. The high levels of thrombomodulin in the chronic-phase cerebral thrombosis group suggests the presence of continuous endothelial cell damage. PMID- 9391720 TI - Inherited abnormalities of blood coagulation in juvenile stroke. A case-control study. AB - The nature of the relationship between inherited abnormalities of the clotting system and the occurrence of cerebrovascular accidents in young subjects is controversial. To evaluate the risk of cerebrovascular disease associated with such abnormalities, we analyzed a series of 23 consecutive patients in a case control study with ischemic stroke proven by computerised tomography and aged below 45 years at admission, and a control group of 115 age- and sex-matched controls from the general population. No differences in antithrombin, protein C, protein S, heparin cofactor II, plasminogen or response to activated protein C were observed between cases and controls. None of the patients had a history of personal or familial thrombosis, and none had a reduction in the considered clotting factor below the reference range. We conclude that abnormalities of the clotting system are not associated with the occurrence of cerebrovascular abnormalities in the young and that routine screening for inherited thrombophilia is not appropriate in young patients with cerebrovascular disease. PMID- 9391721 TI - Insight into the profibrinolytic activity of heparin: effects on the activation of plasminogen mediated by urokinase. AB - The aim of this work was to clarify the role of urokinase-type plasminogen activator (uPA) on the profibrinolytic activity of heparin, chemically modified heparins [partially: N-desulfated (N-des), N-desulfated N-acetylated (N-des N ac), O-desulfated (O-des), O/N-desulfated N-acetylated (O/N-des N-ac)] and heparan sulfate. Binding competition assays of plasminogen and uPA to heparin sepharose demonstrated that heparin bound to both enzymes. Moreover, in the presence of increasing amounts of heparin, plasminogen activation mediated by uPA occurred as a bell-shaped curve, suggesting the formation of a ternary complex. In contrast, all chemically-modified heparins lacked this cofactor activity, although N-des and heparan sulfate partially retained the uPA binding capacity, and O-des partially bound to both plasminogen and uPA. Plasmatic euglobulins from mice treated with heparin, as well as with modified heparins with uPA binding capacity, presented a 2-fold enhancement of 47 kDa lytic band, as assessed by zymographic analysis. Western blotting analysis anti-uPA (47 kDa) showed that the enhanced uPA activity correlated with a true increase in uPA protein levels. These results suggest that the profibrinolytic activity of heparin mediated by uPA could be caused by an increase in uPA protein levels rather than by a cofactor activity mediated by a formation of ternary complexes. PMID- 9391722 TI - Assessment of mental ability in elderly anticoagulated patients: its reduction is associated with a less satisfactory quality of treatment. AB - Mental capacity was assessed in 311 apparently self-sufficient patients (> or = 60 years of age, 170 men) under stabilised oral anticoagulant treatment (OAT) by administering the Hodkinson's Abbreviated Mental Test (AMT). The international normalized ratios (INR) recorded during the 3 months before and the 3 after the data of test administration were examined by the INR-Day software program. The percentage of time spent within, below or above the intended therapeutic range was calculated in patients who scored abnormally at AMT, and compared with matched controls with normal AMT results. Forty patients [12.9%; 28 women (19.8%) and 12 men (7.1%), P < 0.0011] had abnormal AMT results; the rate seemed to increase with age. Most of these patients (35, 75%) had only elementary education. Patients with abnormal AMT results spent more time outside the intended therapeutic ranges than 40 matched controls (20.9% of the observed time vs 13.7%, P < 0.0001; odds ratio 1.68, CI 1.53-1.84). Unsuspected reduction of mental ability or attention levels was found in a number of elderly patients receiving OAT; these patients presented longer periods of either under- or over anticoagulation and were, therefore, exposed to a higher risk of thrombotic or bleeding complications. Anticoagulation clinics would be advised to assess mental abilities in elderly patients before starting OAT. PMID- 9391723 TI - A comparison between continuous infusion versus standard bolus administration of heparin based on monitoring in cardiac surgery. AB - This study was designed to determine prospectively if stable heparin concentrations can be maintained during extracorporeal circulation by using a continuous infusion technique, compared with a bolus regimen based on whole blood heparin concentration monitoring. Forty patients were assigned randomly to either an infusion or a monitoring group. The reference heparin concentration was defined as the whole blood heparin concentration associated with a kaolin activated clotting time (ACT) of approximately 480 s prior to institution of cardiopulmonary bypass (CPB) for both cohorts. For infusion patients, doses of heparin were administered using a continuous infusion based on the initial patient-specific heparin dose per unit weight; heparin was also added to solutions administered after the initiation of CPB based on the reference heparin concentration. For monitoring patients, the dose of heparin administered during CPB was calculated by the Hepcon instrument. Blood specimens collected prior to and during the CPB period were used to measure anti-Xa plasma heparin concentration and complete blood counts, kaolin ACT and whole blood heparin concentration. Doses of heparin and protamine administered and transfusion requirements were similar in patient cohorts. The apparent rate of clearance of heparin from plasma was variable among patients in the monitoring group prior to CPB. Stable heparin concentrations were maintained using whole blood heparin measurements, whereas mean heparin concentrations were slightly lower using the continuous infusion technique. Therefore, an optimal approach might involve the combined use of these regimens. PMID- 9391724 TI - Influence of plasma volumetric errors on the prothrombin time ratio and International Sensitivity Index. AB - The International Sensitivity Index (ISI) for prothrombin time systems depends on the thromboplastin manufacturer's recommended method for use. The purpose of the present study was to investigate the influence of small deviations from the recommended sample volume on the prothrombin time ratio and ISI. Four commercial reagents were studied; three with low ISI and one with high ISI. The effects of volumetric errors on the ISI were used to assess the associated effects on the International Normalized Ratio (INR). The effect of 10% volume error on the INR was not greater than 5%. The effects with the three low-ISI reagents were slightly greater than those with the high-ISI reagent. It is recommended that each laboratory should check the volumes of sample and reagent used for the prothrombin time test. PMID- 9391725 TI - Recurrent venous thromboembolic disease and factor XI concentrate in a patient with severe factor XI deficiency, chronic myelomonocytic leukaemia, factor V Leiden and heterozygous plasminogen deficiency. AB - There are increasing concerns about the potential thrombogenic risks associated with the use of factor XI concentrates. We describe the case of a 49 year-old man with chronic myelomonocytic leukaemia and severe factor XI deficiency (< 1 u/dl), in whom the use of factor XI concentrate appeared to be associated with the development of venous thromboembolic disease. Subsequent investigations revealed the presence of both the factor V Leiden abnormality and heterozygous plasminogen deficiency. This case highlights the risks associated with the use of factor XI concentrates and suggests that these risks may be further increased in patients with an inherited or acquired prothrombotic abnormality or an underlying malignancy. Prothrombotic screening of patients with severe factor XI deficiency may be indicated particularly in younger patients in whom treatment with factor XI concentrates is a possibility. PMID- 9391726 TI - Fibrinogen Poissy I: a new case of the A alpha Arg 16His fibrinogen variant. AB - A fibrinogen variant was identified in a patient with disseminated intravascular coagulation and in one member of her family. Coagulation studies showed marked prolongation of both the thrombin and reptilase times and discrepancy was noted between the levels of plasma fibrinogen, determined by a kinetic vs immunological determination or light scattering assay. Studies on purified fibrinogen revealed an impaired release of fibrinopeptides by thrombin. DNA sequencing revealed a heterozygous A to G point mutation in exon 2 of the A alpha chain, which substituted Arg for His at position 16. This mutation creates a Nla III cleavage site which was used to confirm the mutation. PMID- 9391727 TI - Variant of intron 22 inversions in the factor VIII gene in severe hemophilia A. AB - Recurrent DNA inversions, which disrupt the factor VIII (FVIII) gene, generally occur between a region of intron 22 (int22h) and one of two homologous copies of this region, located 300 to 400 kb telomeric to the FVIII gene. This report describes a patient with severe hemophilia A and a high level inhibitor with atypical hybridization patterns. A Bcl I Southern blot assay was altered to 17.5, 16, and 14 kb. His mother and two out of four aunts tested had normal and abnormal restriction patterns which led to a total of five different fragments, suggesting that they were carriers. The Xba I plus Kpn I restriction fragment length polymorphism in intron 22 by Southern blotting using the same probe (probe a) yielded the 6.2 kb polymorphic band, with a clearly separated 6.6 kb band from the non-factor VIII region; an alternative int22h hybridization probe (probe x) detected no additional fragment. These results suggest that probe a as well as probe x could recognize an intron-22-sized fragment. This report shows a variation in the number of int22h copies although we could not find the inversion junction. PMID- 9391728 TI - Natural anticoagulation with thrombocytopenia may spare the thromboembolic complications in severe cyanotic congenital heart disease. PMID- 9391729 TI - Neural networks for the analysis of small pulmonary nodules. AB - PURPOSE: Small pulmonary nodules can be readily detected by computed tomography (CT). The goal of this detection is to diagnose early lung cancer as the five year survival at this early stage is over 70% in contradistinction to the overall 5-year survival of around 10%. Critical to the efficacy of CT for early lung cancer detection is the ability to distinguish between benign and malignant nodules. We explored the usefulness of neural networks (NNs) to help in this differentiation. METHODS: CT images of 28 pulmonary nodules, 14 benign and 14 malignant, each having a diameter less than 3 cm were selected. All were sufficiently malignant in appearance to require needle biopsy and surgery. The statistical-multiple object detection and location system (S-MODALS) NN technique developed for automatic target recognition (ATR) was used to differentiate between these benign and malignant nodules. RESULTS: S-MODALS was able to correctly identify all but three benign nodules. S-MODALS classified a nodule as malignant because it looked similar to other malignant nodules. It identified the most similar nodules to display them to the radiologist. The specific features of the nodule that determined its classification were also shown, so that S-MODALS is not simply a "black box" technique but gives insight into the NN diagnostics. CONCLUSION: This initial evaluation of S-MODALS NNs using pulmonary nodules whose CT features were very suspicious for lung cancer demonstrated the potential to reduce the number of biopsies without missing malignant nodules. S-MODALS performed well, but additional optimization of the techniques specifically for CT images would further enhance its performance. PMID- 9391730 TI - Giant bronchogenic cyst masquerading as tension pneumothorax. Radiographic and CT findings. AB - Pulmonary bronchogenic cysts with tracheobronchial communication may occasionally mimic tension pneumothorax leading to unnecessary thoracostomy. We describe such a case to emphasize that cautious identification of the direction of displacement of the collapsed lung tissue on chest radiograph or computed tomography (CT) may help in differentiating these two diseases. Tension pneumothorax should lead to centripetal compression of the ipsilateral lung toward the hilum while giant bronchogenic cysts result in centrifugal displacement of the adjacent lung away from the hilum. PMID- 9391731 TI - Gadolinium utilization in the MR evaluation of cardiac paraganglioma. PMID- 9391732 TI - Accuracy of CT in estimating extent of pancreatic necrosis. AB - The objective of this study was to determine whether nonenhancing pancreatic lesions are accurate in estimating pancreatic necrosis. Twenty-six consecutive abdominal computed tomography (CT) examinations performed over a 3-year period that met the CT criteria for pancreatic necrosis were reviewed. Follow-up CTs in three of 26 patients demonstrated pancreatic enhancement, indicating viable parenchyma, within the previously nonenhancing regions. All three patients had undergone surgical debridement in that area. Twenty-three cases demonstrated either no change or enlargement of the nonenhancing pancreatic lesions. Follow-up ranged from 1 week to 26 months. While CT is accurate in diagnosing pancreatic necrosis, lack of enhancement in CT may occasionally overestimate the extent of necrosis. Nonenhancing, viable but at-risk tissue may be present adjacent to frankly necrotic tissue. Surgical debridement may facilitate recovery of this viable tissue, which may enhance normally on follow-up CT. PMID- 9391733 TI - MRI findings in headbangers. AB - Magnetic resonance imaging (MRI) findings in a mentally retarded adult female who exhibits headbanging behavior are presented. Radiographic changes include enlargement of the diploic space in the parietal and occipital bones, and gray matter loss adjacent to the bony changes. This pattern of injury is compared with skull changes previously reported in headbangers, and neuronal injury seen in boxers (dementia pugilistica) and Minimata disease. PMID- 9391734 TI - CT and US of the pancreas. AB - The diagnostic capabilities of pancreatic imaging continue to improve with technological advancements in computed tomography (CT), ultrasound (US), and magnetic resonance imaging (MRI). To update the practicing radiologist, this article summarizes the current literature on pancreatic imaging, with particular emphasis on CT and US. Pertinent clinical considerations of the disease entities are included, along with illustrative material from the authors' experience. PMID- 9391735 TI - Management of asymptomatic term neonates whose mothers received intrapartum antibiotics--Part 2: Diagnostic tests and management strategies. Center of Disease Control and Prevention. AB - The evaluation of the potentially septic newborn is often a source of frustration for practitioners. In the past, it has often been standard practice to evaluate and treat empirically all neonates whose mothers received antibiotics during labor, regardless of whether the infant had any signs or symptoms suggestive of infection. With the advent of recommendations for intrapartum antibiotic therapy to prevent early-onset neonatal group B streptococcal infections, this strategy is no longer practicable because too many infants would thus be evaluated and treated needlessly. This two-part review addresses the issues involved in managing asymptomatic newborns whose mothers received intrapartum antibiotics. Part I, published separately, reviewed the rationale behind strategies for preventing intrapartum transmission of bacterial infection. This final part addresses the evaluation and management of the newborn. A number of diagnostic tests are often used in looking for bacterial infections in the neonate. Unfortunately, none of these is both rapid and reliable. A clinical pathway provided here can serve as a useful guide for the clinician, but uncertainty will always remain. Ultimately, each practitioner must determine the degree of risk or uncertainty that he or she can accept on the basis of clinical experience. PMID- 9391736 TI - The moral challenge of children at risk: protective policies and pediatrics. A report of the Children's Services, Inc. Task Force of Greater Cleveland. AB - Social workers and pediatricians are among the professionals who share a society wide concern with current public policies regarding the placement of children at extreme risk. A healthcare professional may successfully treat a child, only to learn later that this same child was the victim of a tragic incident of domestic violence after returning home. Such events are not uncommon, create considerable frustration for pediatricians, and demand an integrated interprofessional and interdisciplinary response. This report emerged from 6 months of task force dialogue with leaders of children's services programs, healthcare professionals, clergy, ethicists, and other community leaders in one major urban environment. It indicates innovative directions in children's protective services with regard to family preservation, foster care, residential care, and adoption. The latter two options could be used much more creatively than is the case currently. The report also asserts that far too few resources are being directed to this problem area. PMID- 9391737 TI - Using the clinical linguistic and auditory milestone scale for developmental screening in high-risk preterm infants. AB - Eighty-one preterm infants (mean gestational age 29 weeks, range 24-36 weeks) discharged from The Johns Hopkins Hospital Neonatal Intensive Care Unit were followed up sequentially from birth to 2 years of age by use of the Clinical Linguistic and Auditory Milestone Scale (CLAMS) to evaluate language development. Children were studied during three time intervals: Interval 1: 3-5 months chronologic age (CA); Interval 2: 9-14 months (CA); and Interval 3: 18-24 months (CA). Psychometric test scores were compared with CLAMS Language Quotients (LQ) by use of full, partial (75%, 50%, 25%), and no "correction" for weeks of prematurity to determine whether "correcting" for prematurity would yield a more accurate estimate of eventual cognitive outcome. CLAMS LQ at Interval 1 was highly correlated with CLAMS LQ at Interval 2 and CLAMS LQ at Interval 2 correlated well with CLAMS LQ at Interval 3 (r = 0.57 and 0.64, respectively, P = 0.0001). Correlations indicated that there was an orderly, sequential development of language in the preterm infant. CLAMS evaluations correlated significantly with psychometric test results during Interval 2 and Interval 3 (r = 0.34, P < 0.02 and r = 0.75, P = 0.0001, respectively). The CLAMS proved to be a useful instrument for monitoring preterm language development in the primary pediatric care setting. PMID- 9391738 TI - The incidence of acute and remote seizures in children with intraventricular hemorrhage. AB - Seizures are a well-known complication of intraventricular hemorrhage (IVH) in premature infants; however, the rate at which they occur is not known. The authors decided, therefore, to investigate both the incidence of acute and remote seizures in infants with IVH and the association with the grade of hemorrhage. One hundred and three infants with IVH were identified and their records were reviewed for acute seizures, remote seizures, and associated morbidity and mortality. The average gestational age of these infants was 29 weeks (range, 23 40 weeks). Of the 103 infants, 32 (31%) developed grade 4 IVH; 19 (18%), grade 3 IVH; and 52 (50%), grades 1 and 2 IVH. Seventeen (17%) patients had acute seizures during their first month of life. Six of the 61 patients (10%) who survived the neonatal period and for whom follow-up data were available had remote seizures. Infants with grade 4 IVH had significantly more acute seizures than infants with grades 1 and 2. In this cohort, only infants with grades 3 and 4 IVH developed remote seizures. Furthermore, among infants with grade 4 IVH acute seizures were a significant risk factor for development of remote seizures. The use of long-term antiepileptic drug therapy in neonates with a history of acute seizures is not established. These results suggest that antiepileptic drug therapy beyond the neonatal period should be reserved for infants with grade 4 IVH with history of acute seizures. PMID- 9391739 TI - The plantar response in normal newborn infants. AB - The neonatal plantar response has been reported as extensor in 90% of newborns and flexor in 93% of newborns, leading to uncertainty about its reliability and significance. To determine the normal neonatal plantar response we examined 349 healthy newborn infants, > 32 weeks gestation within 24 hours of birth. A supramaximal noxious stimulus was applied in a standardized manner to the lateral plantar surface of each foot. The plantar response was extensor in 90%, equivocal in 7%, and flexor in 3%. With proper physiologic technique, the normal neonatal plantar response is extensor. PMID- 9391740 TI - Bilateral neonatal testicular torsion. PMID- 9391741 TI - Comments on bilateral neonatal testicular torsion. PMID- 9391742 TI - Successful treatment of steroid-resistant chorea associated with lupus by use of valproic acid and clonidine-HCL patch. PMID- 9391743 TI - Endovascular management of vein of Galen aneurysms in neonates: two case reports and a review of the literature. PMID- 9391744 TI - An unusual presentation of a hemangioma in an adolescent female. PMID- 9391745 TI - Size, myths and the clinical pharmacokinetics of analgesia in paediatric patients. AB - In the paediatric population, developmental changes can be predicted by age and are independent of size, which is predicted by bodyweight. Size is commonly standardised using either the per kilogram or the body surface area models. A great many physiological-, structural- and time-related variables scale predictably within and between species with weight exponents of 0.75, 1 and 0.25, respectively. Use of the per kilogram size model has led to the misconception that children have an enhanced capacity to metabolise drugs because of their relatively large liver size or increased hepatic blood flow. This is not necessarily the case. For example, the clearance of opioids often approaches adult rates within the first few months of life when an allometric 3/4 power model is used to scale for size. Age-related changes in physiological processes, such as respiration and cardiac output, disappear with appropriate size models. Size is an important, but little recognised, component in the speed of onset of drugs effects and uptake of inhalational anaesthetic agents. Size models cannot be reliably used to predict dose regimens for children from schedules established for adult patients. Dosage regimens are dependent on clearance and volume of distribution as well as pharmacodynamic factors, which change with age. The age dependent pharmacodynamic changes described for some opioids in the very young have not yet been completely disentangled from age-related pharmacokinetic changes. When bodyweight is standardised and disentangled from age, developmental changes can be understood more clearly. The future investigation of drugs used in paediatric practice must also include an appropriate size model in order to differentiate age-related factors from size-related factors. PMID- 9391746 TI - Pharmacokinetic changes during pregnancy and their clinical relevance. AB - The dynamic physiological changes that occur in the maternal-placental-fetal unit during pregnancy influence the pharmacokinetic processes of drug absorption, distribution and elimination. Pregnancy-induced maternal physiological changes may affect gastrointestinal function and hence drug absorption rates. Ventilatory changes may influence the pulmonary absorption of inhaled drugs. As the glomerular filtration rate usually increases during pregnancy, renal drug elimination is generally enhanced, whereas hepatic drug metabolism may increase, decrease or remain unchanged. A mean increase of 8 L in total body water alters drug distribution and results in decreased peak serum concentrations of many drugs. Decreased steady-state concentrations have been documented for many agents as a result of their increased clearance. Pregnancy-related hypoalbuminaemia, leading to decreased protein binding, results in increased free drug fraction. However, as more free drug is available for either hepatic biotransformation or renal excretion, the overall effect is an unaltered free drug concentration. Since the free drug concentration is responsible for drug effects, the above mentioned changes are probably of no clinical relevance. The placental and fetal capacity to metabolise drugs together with physiological factors, such as differences acid-base equilibrium of the mother versus the fetus, determine the fetal exposure to the drugs taken by the mother. As most drugs are excreted into the milk by passive diffusion, the drug concentration in milk is directly proportional to the corresponding concentration in maternal plasma. The milk to plasma (M:P) ratio, which compares milk with maternal plasma drug concentrations, serves as an index of the extent of drug excretion in the milk. For most drugs the amount ingested by the infant rarely attains therapeutic levels. PMID- 9391748 TI - Renal inflammatory disease: the current role of CT. AB - Computed tomography (CT) plays a significant role in establishing the diagnosis in clinically equivocal cases of renal infection, determining the extent of the disease process, and assessing its complications. Gas, calculi, renal parenchymal calcifications, hemorrhage, and masses can be revealed with unenhanced CT. A subsequent study with contrast enhancement is crucial for the complete evaluation of patients with renal infection in order to demonstrate the areas of altered nephrogram that occur as a result of the inflammatory process and to identify complications. In this article we review a spectrum of renal inflammatory disease, with illustrations of the CT findings in representative cases. We also review the role and potential pitfalls of fast scanning techniques that can image a particular phase of the nephrogram in a renal infection. In acute pyelonephritis, enhanced CT scans obtained during the cortical nephrographic phase typically demonstrate solitary or multifocal hypodense areas with obliteration of the corticomedullary differentiation. Delayed images obtained during the excretory phase are frequently more helpful in defining the extent of the disease process, identifying the complications such as renal abscess, and confirming the presence of urinary obstruction than are early images. PMID- 9391749 TI - MR imaging of the female pelvis: current perspectives and review of genital tract congenital anomalies, and benign and malignant diseases. AB - MR imaging continues to be an integral problem-solving modality in the evaluation of congenital anomalies and acquired diseases of the female genital tract organs and provides effective clinical information to the practicing gynecologist in those patients in whom sonography is technically suboptimal or the results are equivocal. This article describes the state-of-the art MR imaging of the female pelvis and addresses its current perspectives in the following sections: (1) technical aspects of MR in imaging the female pelvis, (2) normal pelvic anatomy and variations that are seen on MRI, (3) role of MRI in the diagnosis of congenital uterine and vaginal anomalies, (4) MR imaging approach to diagnose congenital uterine and vaginal anomalies, (5) advantages and limitations of MR in the evaluation of various benign diseases and malignant neoplasms of the female genital tract, (6) a MR staging system and criteria for each gynecologic malignancy, (7) fundamental MR criteria to differentiate benign from malignant tumors and recurrent tumors from fibrosis, and (8) the present cost-effective value of MR in pregnancy and obstetrics. Magnetic resonance (MR) technology continues to be an important problem-solving modality in the evaluation of benign, malignant, and recurrent diseases of the female pelvic organs with the development of new software and improved hardware over the last few years. The main issues addressed in this article are (1) to review the basic and expanded applications of the current state-of-the art MR imaging in the diagnosis and management of various congenital and acquired disorders of the female pelvic organs, (2) to illustrate a simplified clinico-radiologic (MRI) approach to the diagnosis of congenital and acquired pathologies of the pelvic organs, (3) to provide relevant information to the clinicians to make rational choices among the competing imaging modalities, and (4) to outline the future potential of this modality in the pelvis. PMID- 9391750 TI - Phosphorylation of the proteins of the extracellular matrix of mineralized tissues by casein kinase-like activity. AB - The extracellular matrix of the connective tissue contains non-collagenous proteins (NCP) which are acidic in character. The NCP of mineralizing systems (bone, dentin) differ from those of the non-mineralizing systems (skin, tendon) in that the mineralized tissue NCP are frequently phosphorylated. The phosphorylated proteins have been implicated in various aspects of the mineralization process. Thus, it is of interest to consider the mechanism and regulation of phosphorylation of the major matrix NCP. The majority of the phosphorylation takes place at Ser or Thr residues embedded within acidic sequences, and therefore are targets for casein kinase I (CK1) or casein kinase II (CK2)-like kinases. CK1 and CK2 are distantly related members of the protein kinase family. They are ubiquitous, constitutively active, second-messenger independent kinases. CK1 is found in a variety of isoforms, all homologous to the alpha-subunit of the protein kinase family. It acts as a monomer. The active form of CK2 is a tetrameric holoenzyme, with 2 alpha catalytic subunits and 2 beta regulatory subunits. The CK2 alpha has activity alone, but the holoenzyme is four to five-fold that activity. CK2 can use either ATP or GTP as the phosphate donor, but CK1 can use only ATP. The CK2 activity which phosphorylates the mineralized tissue NCP appears to be localized to membrane-associated cell fractions, and is present in the endoplasmic reticulum and Golgi compartments in osteoblasts, where phosphorylation of the secreted proteins appears to take place as co- and post-translational processes. Data indicate that both alpha and beta subunits of the membrane-associated CK2 are isoforms of the cytosolic CK2 in the same cells. The CK1 has not been specifically localized. Studies of dephosphorylated NCP such as phosphophoryn (PP) have shown that CK1 will not phosphorylate dephosphorylated dPP unless prior phosphorylation with CK2 has been carried out. In turn, CK2 activity may be initiated only after an initial phosphorylation of one of the messenger-dependent kinases. Thus, the phosphorylation reactions in mineralized tissues may be a tightly regulated hierarchical or sequential cascade of intracellular phosphorylation events. PMID- 9391751 TI - Leukocyte adhesion molecules. AB - Recruitment of leukocytes is critical to many of the processes studied in oral biology. With the development of new tools such as monoclonal antibody production and transgenic mice, the specific adhesion molecules thought to be important in leukocyte recruitment have been identified and their function examined. These molecules can be divided into three major classes: selectins, members of the immunoglobulin superfamily, and integrins. They mediate interactions between leukocytes and endothelial cells, facilitating the initial process of leukocyte rolling, firm attachment to endothelium, transendothelial migration, diapedesis, and migration along connective tissue. The goal of this paper is to provide an understanding of which molecules are involved in the above processes by discussing their cellular distribution, counter-receptors, and physiologic function. PMID- 9391752 TI - Models of invasion of enteric and periodontal pathogens into epithelial cells: a comparative analysis. AB - Bacterial invasion of epithelial cells is associated with the initiation of infection by many bacteria. To carry out this action, bacteria have developed remarkable processes and mechanisms that co-opt host cell function and stimulate their own uptake and adaptation to the environment of the host cell. Two general types of invasion processes have been observed. In one type, the pathogens (e.g., Salmonella and Yersinia spp.) remain in the vacuole in which they are internalized and replicate within the vacuole. In the other type, the organism (e.g., Actinobacillus actinomycetemcomitans, Shigella flexneri, and Listeria monocytogenes) is able to escape from the vacuole, replicate in the host cell cytoplasm, and spread to adjacent host cells. The much-studied enteropathogenic bacteria usurp primarily host cell microfilaments for entry. Those organisms which can escape from the vacuole do so by means of hemolytic factors and C type phospholipases. The cell-to-cell spread of these organisms is mediated by microfilaments. The investigation of invasion by periodontopathogens is in its infancy in comparison with that of the enteric pathogens. However, studies to date on two invasive periodontopathogens. A actinomycetemcomitans and Porphyromonas (Bacteroides) gingivalis, reveal that these bacteria have developed invasion strategies and mechanisms similar to those of the enteropathogens. Entry of A. actinomycetemcomitans is mediated by microfilaments, whereas entry of P. gingivalis is mediated by both microfilaments and microtubules. A. actinomycetemcomitans, like Shigella and Listeria, can escape from the vacuole and spread to adjacent cells. However, the spread of A. actinomycetemcomitans is linked to host cell microtubules, not microfilaments. The paradigms presented establish that bacteria which cause chronic infections, such as periodontitis, and bacteria which cause acute diseases, such as dysentery, have developed similar invasion strategies. PMID- 9391753 TI - Mercury exposure from dental amalgam fillings: absorbed dose and the potential for adverse health effects. AB - This review examines the question of whether adverse health effects are attributable to amalgam-derived mercury. The issue of absorbed dose of mercury from amalgam is addressed first. The use of intra-oral Hg vapor measurements to estimate daily uptake must take into account the differences between the collection volume and flow rate of the measuring instrument and the inspiratory volume and flow rate of air through the mouth during inhalation of a single breath. Failure to account for these differences will result in substantial overestimation of the absorbed dose. Other factors that must be considered when making estimates of Hg uptake from amalgam include the accurate measurement of baseline (unstimulated) mercury release rates and the greater stimulation of Hg release afforded by chewing gum relative to ordinary food. The measured levels of amalgam-derived mercury in brain, blood, and urine are shown to be consistent with low absorbed doses (1-3 micrograms/day). Published relationships between the number of amalgam surfaces and urine levels are used to estimate the number of amalgam surfaces that would be required to produce the 30 micrograms/g creatinine urine mercury level stated by WHO to be associated with the most subtle, pre clinical effects in the most sensitive individuals. From 450 to 530 amalgam surfaces would be required to produce the 30 micrograms/g creatinine urine mercury level for people without any excessive gum-chewing habits. The potential for adverse health effects and for improvement in health following amalgam removal is also addressed. Finally, the issue of whether any material can ever be completely exonerated of claims of producing adverse health effects is considered. PMID- 9391754 TI - Tobacco and smoking: environmental factors that modify the host response (immune system) and have an impact on periodontal health. AB - This review summarizes the current data on the effects of smoking and tobacco on the immune system and its potential impact on periodontal health. Smokers are 2.5 6 times more likely to develop periodontal disease than non-smokers, and there is evidence for a direct correlation between the number of cigarettes smoked and the risk of developing disease. Tobacco users also tend to exhibit increased severity of periodontal disease. Direct correlations between tobacco use and increased attachment loss and pocket depth and reduced bone crest height have been reported. Although the correlation between tobacco use and periodontal disease is quite strong, the role of tobacco in the pathogenesis of periodontal disease is uncertain. Recent studies indicate that one potential mechanism is that tobacco use exacerbates periodontal disease because it alters the immune response to periodontal pathogens. Indeed, smokers exhibit increased numbers of peripheral blood mononuclear phagocytes which appear to be functionally compromised. Inadequate phagocyte activity could reduce the clearance of pathogens from the oral cavity and thereby facilitate the development of periodontal disease. Tobacco-exposed B- and T-lymphocytes exhibit reduced proliferative capacities which could limit the production of protective immunoglobulins against oral pathogens. The risk factors for periodontal disease can be broadly classified as genetic, environmental, host-response factors, and host-related factors such as age. Tobacco, an environmental factor, undermines the host response and may facilitate the development and progression of periodontal disease. This review highlights the inter-relatedness of two of the risk factors associated with periodontal disease. PMID- 9391755 TI - Psychosocial factors and secretory immunoglobulin A. AB - This review focuses on studies that have examined the relation between psychosocial factors and secretory immunoglobulin A (s-IgA). Several studies have examined the relation between s-IgA and stressful circumstances ranging from major life events to minor daily events. The findings from these studies were often contradictory, since different experimenters reported different stress related changes in s-IgA. The effects of stress reduction interventions, such as relaxation and imagery, on s-IgA levels have also been examined. Although these studies indicate that various interventions are associated with increases in s IgA levels, methodological refinements are needed before more definitive conclusions can be made. The possibility that the relation between stress and s IgA may be moderated by personality characteristics or mediated by psychological distress was supported in some studies. The review concludes with suggestions for future research. PMID- 9391756 TI - Cystic fibrosis. PMID- 9391757 TI - Early pulmonary disease in cystic fibrosis. AB - In cystic fibrosis, airway infection and inflammation lead to chronic progressive lung disease. The pathogenesis of cystic fibrosis is still not completely understood, but increasing evidence indicates that the disease process occurs in young patients. Treatment of respiratory symptoms in young patients, although not well studied, is commonly accepted and includes the full range of treatments used in older patients-secretion clearance techniques, bronchodilators, anti inflammatory agents, and antibiotics by oral, inhaled, and systemic routes. It is not clear, however, whether early treatment can delay or prevent progressive lung disease in these patients. Outcome measures, including determination of infant lung function, imaging techniques, and direct lower airway sampling through bronchoalveolar lavage are under development and will allow large, multicenter interventional trials in young children. These studies will be aimed at delaying the initiation of lung disease and slowing disease progression. PMID- 9391758 TI - Update on clinical trials of cystic fibrosis. AB - This review attempts to summarize the important areas of cystic fibrosis (CF) clinical research. Some of the trials outlined are incomplete or the data are not yet published. Focus is given to gene therapy and to studies that probe our understanding of CF cellular biology by attempting to correct or bypass abnormal cystic fibrosis transmembrane regulator. Trials of more immediate clinical value include improvement of mucociliary transport and inhaled tobramycin. Finally, mention is made of the significant nonpulmonary treatments for CF including ursodeoxycholic acid for CF liver disease and intracytoplasmic sperm injection for male infertility. PMID- 9391759 TI - The overuse or underuse of dornase alfa. AB - The poor clearance of airway secretions in patients with cystic fibrosis perpetuates the chronic bronchopulmonary sepsis that is predominant. In recent years, novel drugs have been developed to alter the rheologic properties of the secretions in an attempt to improve airway clearance. Dornase alfa reduces the viscoelasticity of sputum from patients with cystic fibrosis and may enhance the clearance of secretions. Current clinical knowledge suggests that it is a safe treatment that improves pulmonary function and reduces respiratory exacerbations. The response, however, is heterogeneous and unpredictable. Scientific studies support the therapeutic rationale for the use of dornase alfa in that treatment reduces the viscoelasticity of airway secretions. Its effect on bacterial persistence and airway inflammation, however, is marginal. The key piece of information that would influence the long-term use of dornase alfa is how it affects disease progression, and at present this is unknown. PMID- 9391761 TI - Ethical considerations in the treatment of cystic fibrosis. AB - Over the last several decades, rapid medical advancement in the treatment of persons with cystic fibrosis (CF) has brought with it a number of ethical concerns. The increasing life expectancy of persons with CF has made transitions in care increasingly common. This change in life expectancy along with advances in assisted reproductive technologies has focused attention on reproductive decision making in CF. Living lobar lung transplantation, although technically developed, remains ethically problematic. The understanding of ethical issues arising in the treatment of CF is crucial to providing optimal care. PMID- 9391760 TI - Choosing a nebulizer for cystic fibrosis applications. AB - As the number of inhaled drugs available for cystic fibrosis grows, there is increasing awareness of delivery device issues. Current jet and ultrasonic nebulizers are inefficient at delivering drugs to the lower respiratory tract. There are large differences in output characteristics between nebulizers and high intersubject variability in lung deposition. The clinical effects of inhaled drugs depend on adequate dosing to the lower airway, so we must choose a nebulizer and patient characteristics that will affect lung deposition positively. Aerosols with particle sizes at the smaller end of the respirable range (2 to 3 microns) may enhance the clinical benefit of some drugs, regardless of patient age or disease severity. Breath-enhanced (Venturi) jet nebulizers are less wasteful than constant-output nebulizers and perform better than conventional nebulizers with many drugs. Patient breathing patterns and degree of airway obstruction are important in determining the site of airway deposition. With increased attention to aerosol delivery issues by clinicians, industry, and regulatory agencies, improved technologies will evolve to target therapies to the lung. PMID- 9391762 TI - Managed care and the cystic fibrosis patient. AB - Since the first published report of mortality in cystic fibrosis in 1969, the median survival among cystic fibrosis patients has risen from 14 to 31 years. The reasons for this improved survival are complex and include earlier diagnosis; improved control of pulmonary infection; aggressive nutritional intervention; and enhanced monitoring of patients in peer-reviewed, accredited centers for cystic fibrosis care, teaching, and research. Emphasis on the importance of research on changing and improved treatment has been effectively communicated to patients and families. As a result, a group of highly educated medical consumers has been created. During the last decade, another focus of rapid change has appeared, that of cost containment in the medical profession, creating the field of managed care. Practicing medical professionals perceive the need for reduction in excessive spending in medicine and have taken a variety of approaches to balance better the value and cost of medical care while maintaining superb quality. Physicians and consumers continue to have concerns about the potential negative impact of managed care on a relatively rare, specialized, and chronic illness such as cystic fibrosis if managed care concepts are applied without proper understanding of the disease. A great concern is that managed care in cystic fibrosis may cause reversal of trends in improved quality and length of survival. The increased length of survival places an increasing demand on the already stressed system of health care financing. Understanding the changing area of managed care is therefore of paramount importance to clinicians involved in cystic fibrosis care. The Cystic Fibrosis Foundation has presented symposia on managed care at each of the last three annual meetings, including the North American Cystic Fibrosis Meeting, October 1996. The following issues were addressed by speakers and panel discussants, with portions excerpted for this review: trends in managed care, measures and guidelines useful in managed care, Medicaid managed care and cystic fibrosis, and practical aspects of using pathways in caring for patients with cystic fibrosis. PMID- 9391763 TI - Early life and the pathophysiology of breathing. PMID- 9391764 TI - New insights into the ontogeny of breathing from genetically engineered mice. AB - Development of breathing behavior depends on the coordinated maturation of central and peripheral neural pathways, respiratory muscles, airways, and lung tissues. Each of these components contains cellular elements in which derangements of gene expression may perturb development of normal respiratory function. Application in recent years of genetic engineering techniques has led to detailed analyses of gene structure and function. In particular, targeted gene deletions provide the opportunity to relate gene function to physiologic mechanisms in intact animals. This review summarizes recent studies in mice designed to alter, by targeted disruption of specific genes, development of individual components of the respiratory control system. We also discuss an example of the human therapeutic potential of transgenic methods. PMID- 9391747 TI - Pharmacokinetic and pharmacodynamic principles of illicit drug use and treatment of illicit drug users. AB - Many clinicians are confronted by the use of illicit drugs on a daily basis. The unsanctioned use of opioids, psychostimulants, benzodiazepines, alcohol and nicotine is a major cause of morbidity and mortality. Multiple factors have inhibited the scientific study of these agents including prohibition, public denial and lack of commercial interests. In dealing with problems related to these drugs, clinicians need a scientific understanding of their pharmacology, quantifiable effects and potential adverse effects. Illicit drug users select drugs with particular pharmacokinetic parameters and pharmacodynamic properties. Generally, rapid absorption, rapid entry into the central nervous system, high bioavailability, short half-life, small volume of distribution and high free drug clearance are pharmacokinetic characteristics which predict a high potential for harmful use because these factors increase positive reinforcement. Drug users adapt the method and route of drug administration to optimise the delivery of the drug to the brain while attempting to maximise the bioavailability of the drug. Inhalation and smoking are the routes of administration which allow the most rapid delivery of drug to the brain, while intravenous injection maximises the bioavailability of an administered drug. Each route of administration results in attendant complications related to mucosal damage, carcinogenesis and risk of infection. Negative reinforcement or withdrawal is a major drive to recurrent use. Many illicit drugs have pharmacological features that promote dependence, including long half-life, low free drug clearance and sufficient drug exposure to allow development of tolerance. The preventive or reductive pharmacotherapeutics of illicit drug use makes use of several subsets of agents: those which act on the same receptor or system as the illicit drug (such as methadone), those which produce an adverse reaction on consumption of the illicit drug (such as disulfiram) and those which symptomatically attenuate illicit drug withdrawal symptoms (such as clonidine). Many new agents are being trialled as potential preventive or reductive agents. It is important to consider pharmacotherapy as only one potential part of the treatment of illicit drug users. The complications of illicit drug use present many therapeutic challenges. As with all patients consuming multiple drugs, illicit drug users are prone to developing drug interactions. The most common interactions seen in practice are pharmacodynamic in nature, most often due to the additive effects of different drugs on the central nervous system. However, alcohol, cocaine, disulfiram, methadone and tricyclic antidepressants may be involved in important pharmacokinetic interactions. Of these the effect of long term alcohol consumption in increasing the hepatotoxicity of paracetamol and of cytochrome P450 3A microsomal enzyme stimulating drugs in diminishing the efficacy of methadone are the most commonly encountered. PMID- 9391765 TI - Development of cardiopulmonary integration and the role of arousability from sleep. AB - The recent literature on cardiopulmonary integration in infants is surveyed here, focusing on arousals from sleep. Recent studies reported that the ontogenicity of cardiopulmonary integration cannot be evaluated independently from that of sleep wake cycles. The issue is of importance for researchers and clinicians evaluating cardiorespiratory characteristics in infants. It also has significant implications in the understanding of clinical conditions, such as parasomnias, obstructive sleep apneas, or some cases of sudden infant death syndrome. The propensity to arouse can be evaluated by exposing the sleeper to awakening challenges. Arousal thresholds are determined by measuring the intensity of the stimulus needed to induce arousals. However, these studies are complicated by factors such as the scoring of the arousal responses. Another difficulty lies in the choice and modality of the arousal stimulus. Noise, gases, light, and nociceptive, mechanical, chemical, or temperature stimuli have all been used to induce arousals. Confounding factors modify the sleeper's responses to a given stimulus. Prenatal drug, alcohol, or cigarette use and the infant's age or, previous sleep deprivation also modify thresholds. Other confounding factors include time of the night, sleep stages, the sleeper's body position, and sleeping conditions. Arousal can also occur spontaneously because of endogenous stimuli. The literature surveyed here also covers such unresolved issues as the clinical significance of aborted arousals, or the mechanisms responsible for the arousal reactions. PMID- 9391766 TI - Pediatric hypoventilation syndromes. AB - Hypoventilation syndromes are an uncommon but important group of respiratory control disorders in infants and children. Congenital central hypoventilation syndrome (CCHS) is the principal and most important example. No specific anatomical or biochemical mechanism has yet been identified. This article summarizes current knowledge regarding CCHS in infants and children, and emphasizes the most recent and most important publications. The most recent advances in CCHS pertain to its genetics, pathophysiology, diagnosis, and treatment and provide state-of-the-art information regarding advances in diaphragm pacing, responses to exercise, and long-term outcome. CCHS is now being recognized more frequently, treatment is more successful, and long-term outcomes are encouraging with timely diagnosis, state-of-the-art treatment, and comprehensive follow-up at an experienced pediatric referral center. PMID- 9391767 TI - Sleep disorders in infancy, childhood, and adolescence. AB - Sleep disorders cause substantial problems during infancy, toddlerhood, preschool ages, school ages, and adolescence. They represent the most common behavioral problems facing most parents, as well as some of the most unusual and fascinating disorders known to medicine. Sleep disorders can result from pulmonary problems, neurologic problems, family problems, or psychologic or psychiatric problems. The majority of these disorders can be diagnosed by a comprehensive sleep and medical assessment, but special studies such as polysomnography, multiple sleep latency testing, or video electroencephalographic monitoring are necessary for certain diagnoses. Pediatric sleep disorders represent a true interdisciplinary and developmental field, richly connected with many aspects of health care and medical science. Physicians and other pediatric care providers must become increasingly knowledgeable about sleep disorders to offer the best care to their patients. PMID- 9391768 TI - Consequences of sleep-disordered breathing in childhood. AB - Obstructive sleep-disordered breathing (SDB) is a common problem in children that may lead to growth failure, neurocognitive and behavioral abnormalities, cor pulmonale, and death. Primary snoring, upper airway resistance syndrome, and obstructive sleep apnea syndrome represent a spectrum of clinical manifestations accompanying increasing degrees of upper airway obstruction. Clearly, children with severe SDB need to be identified and treated promptly. Appropriate management strategies for milder forms of SDB are less clear. Some snoring children, for example, may have an increased frequency of obstructive apnea during sleep, with or without mild hypoxemia, but have essentially no daytime symptoms or apparent clinical consequences. Should these children be treated? If untreated, will these children eventually develop more severe obstructive SDB? Development of management strategies is hampered by the lack of data on the natural history of childhood SDB and on the correlation of specific polysomnographic abnormalities to symptoms and complications. In this review, we highlight recent information about the consequences of obstructive SDB in children, with particular emphasis on areas in which more data are needed. PMID- 9391769 TI - Management of obstructive sleep apnea in childhood. AB - The childhood obstructive sleep apnea syndrome is a common and serious problem. Adenotonsillectomy remains the mainstay of treatment. Nasal continuous positive airway pressure is effective and well tolerated in those who do not respond to adenotonsillectomy. In selected cases, additional surgery or supplemental oxygen (with careful monitoring) may play a role. New guidelines for pediatric polysomnography should help standardize methods, thus enabling a better comparison of outcomes. More research in this area is sorely needed. PMID- 9391770 TI - Cystic fibrosis. PMID- 9391771 TI - Sleep and respiratory neurobiology. PMID- 9391772 TI - NSAIDs and increased blood pressure. What is the clinical significance? AB - Several randomised studies have demonstrated that various nonsteroidal anti inflammatory drugs (NSAIDs) elevate blood pressure in normotensive and hypertensive individuals; however, these data have been contradicted by numerous negative studies. Two meta-analyses have demonstrated that, after pooling data drawn from published reports of randomised trials of younger adults, NSAID use produces a clinically significant increment in mean blood pressure of 5 mm Hg, most marked in patients with controlled hypertension. Stratification by NSAID type revealed that piroxicam, naproxen and indomethacin had the greatest, and sulindac the smallest, pressor effect. These data were supported by 2 large community studies involving elderly patients. Recent NSAID users had a 1.7-fold higher risk of requiring the initiation of antihypertensive therapy compared with nonusers; NSAID users also had a 40% increased risk of receiving a diagnosis of hypertension compared with nonusers. It is vital to determine the nature of the association in the elderly, 12 to 15% of whom are concurrently receiving an NSAID and an antihypertensive agent. Importantly, a 5 to 6 mm Hg increase in diastolic blood pressure maintained over a few years may be associated with a 67% increase in total stroke risk and a 15% increase in coronary heart disease events. While the mechanism(s) remain speculative, salt and water retention through several factors operating in parallel, coupled with increased total peripheral vascular resistance, via increased renal endothelin-1 synthesis, are potentially important. Clinicians should strive to avoid excessive use of NSAID treatment and consider well-tolerated therapeutic alternatives, including simple analgesics and physical therapy. For patients who require concomitant NSAID and antihypertensive treatment, physicians should be aware that indomethacin, naproxen and piroxicam may be associated with a greater pressor effect than many other NSAIDs, and that antagonism of beta-blockers may be greater than that of vasodilators (including ACE inhibitors and calcium antagonists) and diuretics. Finally, the progress of each patient should be monitored by careful blood pressure measurement, particularly during the period of initiation of NSAID therapy. PMID- 9391773 TI - Therapeutic options for HIV-associated bodyweight loss. A risk-benefit analysis. AB - Involuntary bodyweight loss, a common complication of infection with HIV, is an indicator of poor prognosis and decreased survival. Because of the multifactorial pathogenesis of HIV-related wasting, emerging therapies are directed at the multiple proposed mechanisms of involuntary bodyweight loss. The initial evaluation and treatment of HIV-related bodyweight loss is focused on the identification and treatment of reversible causes of bodyweight loss, such as secondary opportunistic infections or endocrine dysfunction. Nutritional intervention should begin in the early stages of HIV infection and continue throughout the life of the patient. Of the appetite stimulants, megestrol most consistently promotes bodyweight gain, but with a predominance of fat, not lean, body mass. Anabolic therapies such as testosterone derivatives and recombinant human growth hormone (somatropin) stimulate the addition of lean body mass and are begin actively researched for the treatment of HIV-associated wasting. Finally, thalidomide, a potent inhibitor of tumour necrosis factor-alpha, is a potentially useful therapy that is still under investigation. New research into the treatment of HIV-related bodyweight loss is focusing on combination therapies. PMID- 9391774 TI - A risk-benefit assessment of growth hormone use in children. AB - Growth hormone prepared by recombinant DNA technology (somatropin) has been commercially available for over 11 years. More than 38,000 children have been treated with different growth hormone products. While the best response to treatment occurs in children with severe growth hormone deficiency, therapy with growth hormone will increase the rate of statural growth in children with short stature of many different aetiologies. There are few studies of the effect of growth hormone treatment of final adult height, and the magnitude of this effect is harder to gauge, particularly in children with idiopathic short stature. Other benefits of growth hormone treatment in children include improvement in psychosocial functioning and physiological parameters, such as bone mineral density. Adverse effects associated with growth hormone treatment have been relatively uncommon. Most of the safety data on growth hormone have come from large postmarketing databases maintained by 2 pharmaceutical companies. The adverse event profile reported in children treated with growth hormone is different from that found in adults. Peripheral oedema and carpal tunnel syndrome, which are common in adults treated with growth hormone and frequently result in treatment discontinuation, are rare in children. Intracranial hypertension is rare, but can occur in children with growth hormone deficiency, Ullrich-Turner syndrome or renal insufficiency during the first 8 to 12 weeks after the start of growth hormone treatment; it has seldom been reported in adults with growth hormone deficiency. Children with growth hormone deficiency, Ullrich-Turner syndrome or renal insufficiency are prone to develop slipped capital femoral epiphyses both before and during growth hormone treatment. Therefore, limping and complaints of hip or knee pain should be carefully investigated. PMID- 9391775 TI - A risk-benefit assessment of octreotide in the treatment of acromegaly. AB - Acromegaly was the first pituitary disease to be recognised as a clinical entity, although initially it was not clear whether the eosinophilic adenomas causing pituitary enlargement were causative or just a manifestation of the syndrome itself. Following the documented clinical improvement of patients with acromegaly after partial hypophysectomy, it was proven that the pituitary adenomas were aetiological. The treatment of acromegaly has changed during the last decades; the introduction of the somatostatin (SMS) analogue octreotide has had major implications. Octreotide was the first SMS analogue to become available for clinical use. It is generally well tolerated, but is associated with the development of gallstones in 15 to 20% of patients. Other adverse effects include transient injection-site pain, abdominal, diarrhoea, gastritis (long term therapy) and loss of scalp hair. No long haematological or biochemical adverse effects have been reported. Desensitisation to the beneficial effects of octreotide therapy is highly unusual. A long-acting formulation of octreotide is being studied, and should be available by the end of 1997. PMID- 9391778 TI - Assessing disease activity in Crohn's disease--are we there yet? AB - Many attempts have been made over recent years to assess accurately disease activity in Crohn's disease. We review some of these attempts, giving particular emphasis to the combination of serum levels of proinflammatory cytokines (interleukin-6, tumour necrosis factor alpha, recombinant interleukin-2 and acid alpha-1-glycoprotein). PMID- 9391776 TI - Heparin-induced thrombocytopenia. Pathogenesis, frequency, avoidance and management. AB - Heparin-induced thrombocytopenia (HIT) is an immunoglobulin-mediated adverse drug reaction associated with a high risk of thrombotic complications. The pathogenic antibody, usually immunoglobulin (Ig) G (HIT-IgG), recognises a multimolecular complex of heparin and platelet factor 4, resulting in platelet activation via platelet Fc receptors. In addition to in vivo platelet activation, it is now recognised that there is a concomitant activation of coagulation, as shown by marked elevations in thrombin-antithrombin complex levels. It is possible that this increased thrombin generation predisposes HIT patients to a newly recognised complication: warfarin-induced venous limb gangrene. This syndrome is characterised clinically by necrosis complicating deep venous thrombosis in the absence of large-vessel arterial occlusion, and appears to result from acquired protein C deficiency during warfarin therapy for deep vein thrombosis and HIT. The recommended treatment for HIT is an agent that reduces thrombin generation, either indirectly via factor Xa inhibition [e.g. danaparoid sodium (a mixture of anticoagulant glycosaminoglycans)] or directly using a specific thrombin inhibitor (e.g. recombinant hirudin; argatroban). HIT is potentially preventable: there is a lower frequency of HIT, associated thrombosis and HIT-IgG seroconversion in patients treated with low-molecular-weight heparins, compared with unfractionated heparin. PMID- 9391777 TI - Traditional remedies and food supplements. A 5-year toxicological study (1991 1995). AB - Since 1991, the Medical Toxicology Unit (MTU) at Guys' Hospital, London, has been assessing the toxicological problems associated with the use of traditional and herbal remedies and dietary supplements. This assessment was carried out by evaluating reports to the National Poisons Information Service (London) [NPIS(L)] which provides emergency information to medical professionals. Relevant telephone enquiries to NPIS(L) were identified. Further case details were obtained by follow-up questionnaire, clinical consultation, toxicological analysis of samples from patients and/or products and botanical identification of plant material. Of 1297 symptomatic enquiries evaluated there was a possible/confirmed association in 785 cases. Case series have been identified which substantiate previous reports, including liver problems following the use of Chinese herbal medicine for skin disorders, allergic reactions to royal jelly and propolis and heavy metal poisoning caused by remedies from the Indian subcontinent. Although the overall risk to public health appears to be low, certain groups of traditional remedies have been associated with a number of potentially serious adverse effects. Considering the extent of use of herbal remedies and food supplements a comprehensive surveillance system for monitoring the adverse health effects of these products is essential. Surveillance of a large population is needed for the complex task of identifying the uncommon and unpredictable adverse effects which are potentially serious. In the UK, the Medicines Control Agency responded to the MTU report by recognising the need for vigilance and by incorporating adverse reactions reporting on unlicensed herbal remedies into their drug reaction monitoring function. As a further step to safeguard the patients/consumers an effective single regulatory system is required which would ensure the safety and quality of all herbal remedies and food supplements available in the UK. PMID- 9391779 TI - Detection of osteoporosis in patients with inflammatory bowel disease. AB - The risk of osteoporosis is increased in patients with inflammatory bowel disease: particularly in those with additional strong risk factors such as glucocorticoid therapy, hypogonadism, past history of fragility fracture or malnutrition. Where possible, bone densitometry should be performed to identify those in need of treatment, to avoid unnecessary treatment if bone density is normal and to monitor the effects of treatment designed to prevent bone loss. If bone densitometry is not available, treatment should be advised on the basis of strong risk factors. Hormone replacement therapy should be given to patients with hypogonadism and bisphosphonate therapy to those receiving long-term glucocorticoid treatment. The dose of glucocorticoids should be kept to a minimum and, where present, vitamin D deficiency should be corrected. PMID- 9391780 TI - Helicobacter pylori and non-steroidal anti-inflammatory drugs: lone agents or partners in crime? AB - A variety of mechanisms are responsible for the gastric and duodenal mucosal injury known to result from the consumption of non-steroidal anti-inflammatory drugs (NSAIDs). Many of these mechanisms may be influenced by coexistent infection with Helicobacter pylori. However, evidence of increased risk from NSAIDs in patients with this bacterium is contradictory. While some authors have reported that symptoms, severity and prevalence of mucosal damage are higher in H. pylori-positive individuals taking NSAIDs than in those who are H. pylori negative, others have noted no significant difference. Reasons for this conflict may include the age of the subjects studied, duration of treatment, toxicity of the NSAID employed and pathogenicity factors related to different strains of H. pylori. PMID- 9391781 TI - A high serum concentration of interleukin-6 is predictive of relapse in quiescent Crohn's disease. AB - BACKGROUND/AIMS: Relapses of Crohn's disease are difficult to predict. We assessed the value of serum level of interleukin-6, tumour necrosis factor alpha (TNF-alpha) and soluble TNF receptors as predictors of relapse in quiescent Crohn's disease. PATIENTS/METHODS: Thirty-six patients with inactive Crohn's disease, treated or not, were included. Various clinical and biological parameters, including interleukin-6, TNF-alpha and soluble TNF receptors serum levels were measured at inclusion in the study and the patients were followed clinically for 1 year. The relapse was defined as a Crohn's Disease Activity Index (CDAI) greater than 150 with an increase greater than 100 compared to the inclusion value. We analysed the ability of these parameters to predict relapse in parallel to clinical characteristics and other laboratory parameters. RESULTS: Among the 32 variables tested, interleukin-6 serum level had the greatest ability to predict the time-to-relapse, with 17-fold chance of relapse over a 1-year period for patients with an interleukin-6 serum level greater than 20 pg/ml than for patients with a lower level (P < 0.001). A high serum level of the soluble TNF receptors p55 and p75 also had significant predictive value, in contrast to TNF-alpha serum levels. An interleukin-6 serum level greater than 20 pg/ml and either an acid alpha-1-glycoprotein level greater than 1.1 g/l or a soluble interleukin-2 receptor serum level greater than 95 pM/l were risk factors selected by a stepwise multivariate analysis. In both models a good prognosis group was defined by the absence of the two risk factors, a bad prognostic group by the presence of the two risk factors and an intermediate in between. With both models, the good prognosis group included 17 patients who experienced no relapse over the 1-year follow-up, whereas all patients (seven with the first model and six with the second) in the bad prognosis group had a relapse during the follow up. Looking specifically at two homogeneous subgroups including either naturally/5-aminosalicylic acid (5-ASA) quiescent or corticoid quiescent patients, a very good predictive value for interleukin-6 serum concentration was also found. CONCLUSION: Interleukin-6 serum level alone or in association with other biological parameters such as acid alpha-1-glycoprotein or the soluble interleukin-2 receptor serum level may be useful for predicting the course of the disease in patients with quiescent Crohn's disease. PMID- 9391782 TI - The relation of hand skin-fold thickness to bone mineral density in patients with Crohn's disease. AB - OBJECTIVES: In healthy postmenopausal women, the association of skin-fold thickness (SFT) with bone mineral density (BMD) is well described, and a low SFT is a useful predictor of osteoporosis. In this study the association between hand SFT and BMD in patients with Crohn's disease was assessed; and the potential for hand SFT as a screening test for osteoporosis evaluated. DESIGN/METHODS: In a cross-sectional study, BMD was measured at the hip and lumbar spine by dual energy x-ray absorptiometry (DEXA). SFT was measured on the dorsum of the right hand using Holtain Tanner Whitehouse calipers. One hundred and seventeen patients (48 male) with Crohn's disease and 50 (25 male) controls were studied. RESULTS: There was a significant correlation between hand SFT and BMD (expressed as t scores) at all four measured sites (lumbar spine r = 0.41, P < 0.0001, 95% CI 0.25-0.55, Ward's triangle r = 0.38, P < 0.0001, 95% CI 0.21-0.53, trochanter r = 0.33, P < 0.0001, 95% CI 0.16-0.48, femoral neck r = 0.38, P < 0.0001, 95% CI 0.21-0.53). On stepwise regression analysis, the association remained significant after correcting for age, weight, menstrual status and current steroid use (P < 0.05). Hand SFT was significantly lower in patients with Crohn's disease than controls (difference in means 0.51 mm, 95% CI 0.3-0.72, P < 0.0001). Mean hand SFT was significantly lower in patients with osteoporosis compared to patients with normal BMD (difference in means 0.74 mm, 95% CI 0.33-1.15, P < 0.001), as was that of osteopenic patients compared to patients with normal BMD (difference in means 0.28 mm, 95% CI 0.01-0.55, P < 0.05). In the diagnosis of osteoporosis, the sensitivity of hand SFT ranged from 29% to 93%, with specificities of 54% to 95%. CONCLUSIONS: Hand SFT is independently associated with BMD in Crohn's disease and is lower than in age-matched healthy subjects. Hand SFT in combination with other easily measurable confounding variables might be useful in screening for osteoporosis in patients with Crohn's disease. PMID- 9391783 TI - The effect of Helicobacter pylori infection on NSAID-related gastroduodenal damage in the elderly. AB - OBJECTIVE: To evaluate the effect of Helicobacter pylori infection on the prevalence and severity of non-steroidal anti-inflammatory drug (NSAID)-related upper gastrointestinal lesions in the elderly. PATIENTS AND METHODS: One hundred and twenty-eight symptomatic NSAID users (47 males, 81 females; mean age 79.5 years, range 67-95 years) were evaluated by endoscopy. NSAID use was evaluated at the time of endoscopy by interview and general practitioners' clinical records. Patients were separated by the following use patterns: (1) Occasional Users: patients who had taken NSAIDs sporadically, on an as-needed basis in the 7-day period before endoscopy; (2) Acute Users: patients who had taken NSAIDs regularly during the last month; and (3) Chronic Users: patients who had taken NSAIDs regularly for more than 1 month. H. pylori infection was diagnosed by gastric histology (modified Giemsa stain) and the rapid urease test. Statistical analysis was performed by means of the chi 2 test with standardized deviates. RESULTS: Of the 128 subjects, 107 (83.6%) presented with gastroduodenal damage: 3 patients (2.3%) had erosive oesophagitis; 38 patients (29.7%) had gastric ulcer (GU); 43 patients (33.6%) had duodenal ulcer (DU); 3 patients (2.3%) had both GU and DU and 20 patients (15.6%) had erosive gastritis. Seventy-four of the 128 patients (57.8%) were found to be H. pylori positive. In comparison to H. pylori-negative subjects, those who were H. pylori-positive had a significantly higher percentage of GU and DU (74.3% vs. 53.7%, P = 0.02) and a lower percentage of non gastroduodenal lesions (10.8% vs. 24.0%, P = 0.05). No significant trend in H. pylori positivity was found in the 50/128 (39.06%) patients who presented with bleeding lesions (H. pylori positive 36.5%, H. pylori negative 42.6%, not significant). At endoscopy 78% of occasional NSAID users, 93.8% of acute users and 88.7% of chronic users presented with upper gastrointestinal lesions (not significant). No significant differences in NSAID use patterns were observed between H. pylori-positive and H. pylori-negative subjects. CONCLUSION: H. pylori infection in the elderly is associated with an increase in the NSAID-related GU and DU, but not with a higher prevalence of upper gastrointestinal bleeding. PMID- 9391784 TI - Erosive duodenitis: prevalence of Helicobacter pylori infection and response to eradication therapy with omeprazole plus two antibiotics. AB - OBJECTIVES: To study the prevalence of Helicobacter pylori infection in patients with erosive duodenitis (ED), the associated gastric histological lesions and their response to eradication therapy with omeprazole plus two antibiotics. METHODS: A prospective study was made of 57 patients with ED (mean age 46 +/- 16 years, 72% males). At endoscopy, biopsies from gastric antrum and body were obtained for histological study (haematoxylin and eosin). A 13C-urea breath test was also performed. Omeprazole 20 mg twice daily plus two antibiotics (amoxycillin 1 g twice daily, clarithromycin 500 mg twice daily, metronidazole 500 mg twice daily) were administered for 1 week. Endoscopy and breath test were repeated 1 month after completing therapy, and the breath test was performed again at 6 months. RESULTS: All patients were H. pylori positive. Overall eradication was achieved in 86% (95% CI 75-93%). Duodenal erosion healing was obtained in 45 patients (79%). Healing was achieved in 86% (CI 73-93%) of cases with successful eradication therapy, but only in 3/8 (37%; CI 8.5-75%) patients with therapy failure (P < 0.01). In the multivariate analysis, H. pylori eradication was the only variable which correlated with erosion healing (odds ratio 10; CI 2-51; P < 0.01). Histological improvement, in both the gastric antrum and body, was demonstrated when eradication was achieved (P < 0.001). Six months after diagnosis H. pylori absence was confirmed in all patients with initial therapy success (all of them asymptomatic), and infection was confirmed in the eight patients who were H. pylori positive after therapy (six of them symptomatic). At 6-month follow-up, endoscopy was normal in 6/7 H. pylori negative patients with previously persistent ED, while erosions were still present in 4/5 H. pylori-positive patients with previously persistent ED. CONCLUSION: A high prevalence (100%) of H. pylori infection in patients with ED was observed. A 1-week twice daily therapy with omeprazole plus two antibiotics (clarithromycin plus amoxycillin or metronidazole) was very effective in H. pylori eradication, duodenal erosion healing, symptomatic improvement, and in disappearance of associated histological gastritis. These observations suggest that ED should be considered a variant form of duodenal ulcer disease and treated accordingly; that is, with H. pylori eradication therapy. PMID- 9391785 TI - No evidence of activated blood coagulation in Crohn's disease. AB - BACKGROUND: Thromboembolism seems to be a significant and serious complication in Crohn's disease (CD), and multifocal microvascular infarction of the intestinal mucosa may be an important effector mechanism in the pathogenesis of CD. Therefore, it has been hypothesized that an increased activation of the blood coagulation system may favour thromboembolic complications. OBJECTIVES: To assess the activity of blood coagulation as a potential index of thromboembolic risk in CD using thrombin-antithrombin III complex (TAT). DESIGN: Prospective evaluation of TAT. SETTING: Out-patients at the gastroenterological department of a university hospital. PATIENTS: Eighty patients with CD, 47 with inactive (Crohn's disease activity index (CDAI) < 150) and 33 with active disease, and 80 healthy controls were investigated in this study. METHODS: TAT and fibrinogen were used as parameters of blood coagulation. C-reactive protein and orosomucoid were used as serum inflammatory parameters. RESULTS: Fibrinogen was significantly higher in patients with active CD (median 535 mg/dl; interquartile range 402-620 mg/dl) than in patients with inactive CD (357 mg/dl; 300-467 mg/dl) or controls (268 mg/dl; 231-299 mg/dl). Fibrinogen correlated with CDAI, C-reactive protein and orosomucoid. TAT did not show any difference between patients with active CD (3.2 ng/ml; 2.5-4.6 ng/ml), inactive CD (3.0 ng/ml; 2.4-3.9 ng/ml) and controls (3.1 ng/ml; 2.3-3.6 ng/ml). Correspondingly, TAT correlated neither with serum inflammatory parameters and CDAI nor with fibrinogen. CONCLUSION: We could not find evidence of activation of the blood coagulation system as determined by TAT plasma levels in CD, not even in patients with active disease. TAT is not, therefore, a potential index of thromboembolic risk in CD and of microvascular infarction as an effector mechanism in the pathogenesis of CD. PMID- 9391786 TI - Transjugular intrahepatic portosystemic shunt: a limited role in refractory ascites. AB - OBJECTIVE: To evaluate the role of the transjugular intrahepatic portosystemic shunt (TIPS) in the management of patients with refractory ascites. DESIGN: A retrospective study of 25 consecutive patients for whom refractory ascites was the primary indication for TIPS insertion. SETTING: Regional liver unit at Freeman Hospital, Newcastle upon Tyne, UK. PARTICIPANTS AND INTERVENTIONS: Twelve male and 13 female patients with a mean age of 58 years and mean Child-Pugh score of 10, treated with TIPS for refractory ascites between July 1992 and September 1995. MAIN OUTCOME MEASURES: Effect of TIPS on mortality, ascites and hospital admission rate. RESULTS: TIPS was successfully placed in all patients with a 59% mean reduction in portosystemic pressure gradient. Response rate was 68%, 48% and 33% at 1, 3 and 12 months, respectively. Mortality was 48% at 3 months and 67% at 12 months, being higher in those patients older than 60, those with renal impairment and those with higher Child-Pugh score. Amongst nine patients surviving long term (> 12 months) the mean time spent in hospital in the 3 months before TIPS was 35 days and in the year following TIPS 30 days. Patients who died (16 in total) spent a mean of 19 days in hospital before TIPS, 10 never leaving hospital, and 6 who were discharged spent a mean of 19 days post procedure in hospital (mean survival 84 days). CONCLUSION: TIPS has a limited role in the management of patients with refractory ascites. It is not an appropriate treatment where patients are older than 60, have renal impairment (creatinine > 200 mumol/l) or have a Child-Pugh score greater than 10. PMID- 9391787 TI - Relationship between cytochrome P-450 induction by rifampicin, hepatic volume and portal blood flow in man. AB - OBJECTIVE: Induction of hepatic cytochrome P-450-dependent oxidative metabolism is related to an almost identical increase (30%) in both the liver weight and portal blood flow in animals. In humans by contrast, an increased liver blood flow (44%) but no significant increase in liver volume has been reported. DESIGN: Therefore, we studied prospectively the relationship between P-450 induction by rifampicin, hepatic volume and portal blood flow in 10 healthy volunteers. METHODS: After a pre-treatment phase (day 1 to 7) the 10 volunteers received 600 mg/day of rifampicin from day 7 to 12. The urinary 6-beta-hydroxycortisol output as a measure of oxidative metabolism (CYP3A4) and portal blood flow (pulsed Doppler ultrasound) were determined on days 1, 7, 11 and 13. Hepatic magnetic resonance volumetry was performed on days 1 and 13. RESULTS: Urinary 6-beta hydroxycortisol output increased in all volunteers (P = 0.0051) from a median of 2.15 micrograms/day/kg (1.8-3.3 micrograms/day/kg) on day 1 to 9.9 micrograms/day/kg (5.7-14 micrograms/day/kg) on day 13. In 9 of 10 volunteers induction by rifampicin was related to an increase (P = 0.0218) in liver volume from a median of 1570 cm3 (1390-1830 cm3) to a median of 1690 cm3 (1420-1860 cm3). The portal flow as assessed by colour Doppler ultrasound did not change significantly between day 1 (median 22 cm/s (15-35 cm/s)) and day 13 (median 19 cm/s (16-39 cm/s)). CONCLUSION: A fourfold increase of urinary 6-beta hydroxycortisol output after induction of cytochrome P-450 by rifampicin is associated with a significant but less than 10% increase in human liver volume. No increase of portal perfusion as assessed by Doppler ultrasound could be detected in this study. PMID- 9391788 TI - Natural history of advanced hepatocellular carcinoma in Crete. Association with hepatitis C virus. AB - OBJECTIVE: To investigate the clinical characteristics of advanced hepatocellular carcinoma (HCC) in Crete and to analyse the natural course of the untreated disease. PARTICIPANTS: Seventy-three patients (62 men) were enrolled in a prospective 4-year study. Clinical and virological parameters were recorded. Diagnosis was based on either ultrasound guided liver biopsy or a pathognomonic increase in alpha-fetoprotein plus compatible imaging. METHODS: Statistical analysis was performed using histograms, contingency tables and one-way analyses of variance to analyse the characteristics of the disease. For survival analysis Kaplan-Meier survival curves and Cox's proportional hazards models were constructed. RESULTS: HCC in Crete is a mostly male disease (7:1 male:female ratio) and unlike in mainland Greece, it is mostly a hepatitis C virus (HCV) related disease (54% HCV positive as opposed to only 13% in mainland Greece). Prognosis was associated with Okuda classification (Okuda stage III patients have a relative risk of dying that is seven to nine times higher than for Okuda stage I), the presence or absence of hepatitis Be antigen (HBeAg) and antibody to hepatitis B core antigen (anti-HBc). By contrast the presence of anti-HCV was not associated with a worse prognosis. A unit increase of albumin concentration was associated with an 11% decrease in the hazard rate. CONCLUSION: In general, Crete, despite the extremely similar population to the rest of Greece, resembles more closely the situation in Spain or Italy rather than mainland Greece. PMID- 9391789 TI - Platelet function in cirrhosis and the role of humoral factors. AB - BACKGROUND: Haemorrhagic complications are common in patients with cirrhosis of the liver and contribute to the morbidity and mortality seen in this condition. Several pathological processes are involved in these complications, including a delay and overall reduction in platelet aggregation. OBJECTIVE: To determine whether impaired aggregation in cirrhosis is the result of an intrinsic abnormality in platelet function or is induced by a factor present in the blood of cirrhotic patients. SETTING: Liver unit patients in a teaching hospital. DESIGN: Blood was taken from 11 patients with cirrhosis (Child's B or C) and 11 healthy controls. Crossover experiments were carried out, suspending platelet pellets from patients in platelet-poor plasma from controls, and vice versa. Aggregation of both samples and also of platelet-rich plasma from patients and controls was measured. RESULTS: Aggregation after 1 min was impaired significantly in patient, compared with control, platelets following incubation in either patient (P = 0.0012), or control (P = 0.0242), plasma. Correspondingly, maximum aggregation of patient platelets was also reduced significantly (P = 0.0036) after incubation in patient plasma. Aggregation after 1 min, and maximum aggregation, of control platelets was increased significantly (P = 0.034 and 0.013, respectively) following incubation in patient plasma. Furthermore, there was a trend (non-significant) towards reduced aggregation of patient platelets incubated in control plasma. CONCLUSION: These results confirm both an intrinsic platelet defect causing impaired aggregation and a circulating factor in the plasma of patients with cirrhosis which may compensate for this functional defect. PMID- 9391790 TI - Cancer of the pancreas: from concordant classification to standardized treatment. International Symposium at Kloster Irsee, Germany, 2-5 October 1996. AB - There has been a slow but steady improvement in the results of treatment for exocrine pancreatic cancer in the last decade. One obstacle to further improvement has been lack of standardization for reporting of results, from classification to surgery and follow-up. This four-day symposium was organized in order to bridge the clinical praxis of curatively intended radical pancreatic surgery to scientific evaluation of its results. The meeting comprised four workshops, three special lectures, and seven plenary sessions. PMID- 9391791 TI - Hyperkalaemia and diarrhoea in a patient with surreptitious ingestion of potassium sparing diuretics. AB - We report a patient who presented with the unusual combination of chronic diarrhoea and hyperkalaemia. The patient was admitted to our hospital after repeated negative evaluations elsewhere including exploratory laparotomy. The patient had a long history of diarrhoea with hypokalaemia which was documented on several occasions in the past. Several months before admission to our hospital for evaluation of diarrhoea the patient developed hyperkalaemia. Her daily stool output reached 1200 g and her serum potassium was as high as 6.0 mmol/l. Extensive evaluation revealed surreptitious ingestion of the diuretics triamterene, hydrochlorothiazide and spironolactone as the cause of hyperkalaemia and diarrhoea. In addition, she had melanosis coli which was interpreted to be the consequence of surreptitious ingestion of anthraquinone-containing laxatives in the past although no current laxative intake could be proven. We postulate that diarrhoea in our patient was mainly due to the decreased sodium absorption in the small intestine and colon caused by diuretics. Serum aldosterone levels were more than eight times the upper limit of normal. Increased aldosterone levels presumably arose secondary to volume contraction and sodium chloride depletion, but presumably were not able to affect renal and colonic electrolyte transport because of blockage of mineralocorticoid receptors by spironolactone. Thus, the unusual combination of diarrhoea and hyperkalaemia resulted. PMID- 9391792 TI - Diffuse abdominal pain, nausea and vomiting due to retroperitoneal fibrosis: a rare but often missed diagnosis. AB - Retroperitoneal fibrosis is a rare chronic inflammatory disease usually involving the ureters, retroperitoneal vessels and nerves; however, any intestinal organ may also be involved. In recent years, a few successful immunosuppressive treatments of this disease have been described and surgery can, therefore, probably be avoided in most cases. We report here on a case of secondary retroperitoneal fibrosis with compression and midline deviation of the ureters and impaired renal function which was probably caused by ergotamine abuse because of migraine. The patient complained of diffuse abdominal pain, nausea and vomiting. After immunosuppressive treatment with azathioprine and prednisone for a year, we observed a complete resolution of the retroperitoneal mass on computed tomography, although renal function remained impaired. Eleven months after the cessation of treatment, the patient had not relapsed. PMID- 9391793 TI - Palliation of a malignant gastrocolic fistula by endoscopic human fibrin sealant injection. AB - We report the case of a female patient with Hodgkin's disease resistant to therapy who developed a gastrocolic fistula as a consequence of her disease, leading to distressing faeculent vomiting. This was not considered to be amenable to surgical resection and her symptoms were successfully palliated endoscopically using injection of human fibrin sealant into the gastric and colonic aspect of the fistula tract. Both mechanical sealing and promotion of healing by human fibrin sealant are likely to be responsible for its efficacy. PMID- 9391794 TI - Post-cholecystectomy transient hundred-fold increase in CA 19-9. PMID- 9391795 TI - Helicobacter pylori infection and acid secretion. PMID- 9391796 TI - Slide presentations: think about colour blindness. PMID- 9391797 TI - Survey of spinal therapeutic procedures in the United Kingdom. AB - The type and frequency of spinal therapeutic work being undertaken in the United Kingdom (UK) by clinicians with an interest in the surgical treatment of disorders of the spine (primary and secondary subspecialty interest) were evaluated by means of a postal questionnaire. The willingness of respondents to take part in postgraduate spinal training was determined along with issues regarding accessibility of spinal services to non-specialist physicians in the health service in the UK. The results of 450 respondents provided insight into the types of procedures taking place, for example: primary spinal decompression was regularly carried out by 76% of surgeons, while at least 20% of respondents regularly carried out 66% of the procedures surveyed. We found that 10% of surgeons indicated that they were prepared to participate actively in postgraduate spinal surgical training. PMID- 9391798 TI - Cavernous angiomas of the spinal district: surgical treatment of 11 patients. AB - Cavernous angiomas, also called cavernous malformations or cavernomas, are vascular hamartomas accounting for 3-16% of all angiomatous lesions of the spinal district. Although histologically identical, these vascular anomalies may exhibit different clinical behavior and radiological features, depending on their location, hinting at different managements and therapeutic approaches. The authors report 11 cases of symptomatic spinal cavernous angiomas diagnosed and surgically treated over the past 18 years. Age of patients ranged from 15-75 years; males outnumbered females. Three patients had vertebral cavernous malformations, secondarily invading the epidural space; two had pure epidural lesions; two patients had intradural extramedullary lesions, and four intramedullary lesions. Surgical removal was completely achieved in four patients with intramedullary lesions, in two with subdural extramedullary lesions, and in one with a pure epidural lesion. Subtotal excision of another one epidural and three vertebral cavernous angiomas was followed by radiotherapy. There was no morbidity related to surgery; the mean follow-up was 2 years. The outcome was excellent in two cases, good in six, and unchanged in the other three. The authors discuss the different modalities of treatment of these vascular lesions variously placed along the spine. PMID- 9391799 TI - Pregnancy and delivery in patients operated by the Harrington method for idiopathic scoliosis. AB - The course and outcome of 142 pregnancies in 146 patients operated between 1970 and 1975 by the Harrington method for idiopathic scoliosis were studied to determine the effects of scoliosis on pregnancy and childbirth and the effects of pregnancy on the remaining fused and unfused scoliotic curvatures. Occurrence of and sick leave due to low back pain during pregnancy was determined. The patients, all originally treated at the Orthopaedic Hospital of the Invalid Foundation (Orton) in Helsinki, Finland, were invited to a clinical and radiological re-examination on average 19 years following surgery. The results show that pregnancy does not significantly increase fused scoliotic curvatures nor the remaining unfused curvatures. A somewhat higher proportion of children (23%) were delivered by cesarean section than in the general population (15%; P < 0.01), but this result should only be taken as suggestive. Rates of complications of pregnancy and in labor did not differ from those in the background population. The offspring were healthy. Low back pain during pregnancy occurred in about 40% of our patients, but was severe enough to cause sick leave only in 11% of the pregnancies. PMID- 9391800 TI - Quantitative assessment of the motion of the lumbar spine in the low back pain population and the effect of different spinal pathologies of this motion. AB - There are few objective means by which disability caused by low back pain (LBP) can be quantified. The purpose of this study was to investigate the usefulness of motion measurements in the assessment of LBP. The motion characteristics of 138 LBP subjects were investigated, and the data compared with a previously published database of normal subjects. Values of range of motion and angular velocity were obtained for all subjects in each plane of motion. Analysis of these motion characteristics demonstrated significant differences (P < 0.0001) between the two populations; however both populations demonstrated considerable intersubject variation. Multiple regression analysis revealed that some of the variance in the LBP population was attributable to the underlying diagnosis. Patients with a spondylolisthesis tended to be hypermobile whilst those with spinal stenosis, disc prolapse or degenerative disc disease tended to be hypomobile. All diagnostic groups showed impairments in their velocity characteristics. PMID- 9391801 TI - Intraoperative control by somatosensory evoked potentials in the treatment of cervical myeloradiculopathy. Results in 210 cases. AB - Somatosensory evoked potentials (SEPs) were used for continuous monitoring of 210 patients during anterior surgery for cervical myeloradiculopathy, to test how effectively they help avoid irreversible neurological damage during surgery. The pathologies differed in severity and were treated by diskectomy or by extended corporectomy using the Senegas technique. Intraoperative SEP changes were recorded in 84 patients (40%); in 13 (6.2%) of these, changes in SEP amplitude and latency were caused by mechanical stress. SEPs revealed transient episodes of regional ischaemia or neurophysiological anomalies during anaesthesia (mainly hypotension) in 27 patients (12.8%). The traces detected incipient and potentially dangerous mechanical pressure on, or metabolic anomalies of, the spinal cord during manipulation and placement procedures of spinal fixation devices. They were particularly sensitive indicators of ischaemia; one of the most common causes of irreversible injury. The traces of 44 patients (21.0%) improved markedly during surgery. There were no false-negatives in this series and, thanks to the fact that SEPs gave immediate warnings of incipient ischaemia to the surgical team, we had no case of irreversible medullary or nerve-root deficit. PMID- 9391802 TI - Early complications of spinal pedicle screw. AB - The complications of 648 consecutively inserted Universal AO pedicle screws (140 in the thoracic spine and 508 in the lumbar spine) performed by one surgical team to treat 91 patients with spinal problems, were reviewed. The spinal pathology consisted of: scoliosis (34 patients), degenerative lower lumbar spinal disease (25 patients), neoplastic spinal disease (11 patients), thoracic kyphosis (8 patients), spinal fractures (7 patients), lumbo-sacral spondylolisthesis (3 patients), and osteomyelitis (3 patients). Intraoperative complications were: screw misplacement (n = 3), nerve root impingement (n = 1), cerebrospinal fluid leak (n = 2) and pedicle fracture (n = 2). Postoperative complications were; deep wound infection (n = 4), screw loosening (n = 2) and rod-screw disconnection (n = 1). The conclusion was that pedicle screw fixation has an acceptable complication rate and neurological injury during this procedure is unlikely. PMID- 9391803 TI - Aseptic spondylitis as the initial manifestation of the SAPHO syndrome. AB - We describe the case of a 61-year-old female patient who presented with spondylitis of the lumbar spine. Although the microbiological cultures of the bone biopsy specimens obtained during laminotomy remained negative, the patient was treated with broad-spectrum antimicrobials for 2 months. Eight months later she started to suffer from pain and tenderness in her sternum and the medial portion of her left clavicle. The findings of computed tomography and gallium labelled isotope scan were indicative of sternoclavicular arthritis. Again, all surgically obtained biopsy specimens yielded negative results in microbiological studies. The diagnosis of the SAPHO (synovitis, acne, pustulosis, hyperostosis, and osteomyelitis) syndrome was then made based on the clinical presentation with recurrent sterile osteitis in two characteristic locations, and the patient was started on immunosuppressive therapy. This case is a reminder that SAPHO may sometimes occur without any skin manifestations. Since this type of patient may be admitted to an orthopedic ward, it is important that orthopedic surgeons are familiar with the syndrome. PMID- 9391804 TI - Tuberculosis of the sacroiliac joint. AB - A 23-year-old woman presented with low back pain of several months' duration. A tuberculous infection of the left sacroiliac joint was diagnosed by closed-needle biopsy. The clinical presentation, radiological features and outcome of this patient are discussed. PMID- 9391805 TI - Surgical decompression: a life-saving procedure for an extensive spinal epidural abscess. AB - Extensive spinal epidural abscesses (SEAs) carry a high mortality rate. Traditionally they are treated non-operatively with long-term antibiotics and/or surgical decompression, but there is a continuing debate as to whether they should be managed by emergency surgical decompression. However, such decisions are made in the light of the clinical setting. We report the successful management of a female patient who presented with features of upper cervical cord compression and later developed septic shock and multisystem failure. Surgical decompression of the cervical spine and irrigation of the epidural space with a paediatric catheter was performed followed by tricortical strut grafting and plating. At review, 36 weeks after surgery, the patient remained asymptomatic, having made full neurological recovery. The purpose of this report is to highlight the importance of emergency surgical intervention for extensive SEA in the presence of progressive neurological loss associated with multisystem failure. PMID- 9391807 TI - Anteriorly displaced transverse fractures of the sacrum in adolescents: report of two cases. AB - A fracture of the sacrum at the level of the first and second segments, with forward displacement of the first segment, is a very rare injury in adolescents. The cases of two patients, who both suffered a displaced transverse fracture of the sacrum with resulting neural disturbance, are reported here. We consider that these unstable fractures may be treated surgically, by extensive laminectomies of the lumbosacral area and posterolateral fusion. Stabilization of the displaced fracture is possibly preferable, because it provides the prerequisites for early mobilization and reduces pain. PMID- 9391808 TI - Fracture of the vertebral limbus. AB - An 18-year-old young man suffering from fracture of the limbus of L4 was admitted to the emergency ward after a car collision. Radiological evidence of the lesion was visible on plain film radiographs and CT scans. On surgery the posterior column was found to be intact. Treatment included a wide laminectomy, excision of the fragment, and osteosynthesis with Cotrel-Dubous-set instrumentation. The characteristics of these lesions are reviewed on the basis of the latest reports. The possibility of misreading these fractures is emphasized, especially in traumatic adult spine surgery. PMID- 9391806 TI - Iliopsoas bursitis and pseudogout of the knee mimicking L2-L3 radiculopathy: case report and review of the literature. AB - We report the case of a 74-year-old woman who presented with acute-onset right groin pain irradiating to the thigh anteriorly after having suffered for a few weeks from slight knee pain. As a CT scan showed multiple herniated intervertebral discs and spinal stenosis at the L3-L4 level, she was referred to a neurosurgical unit with the tentative diagnosis of L2-L3 radicular pain. Investigations (MR, myelography with CT scan) showed severe acquired lumbar canal stenosis. Decompression surgery was finally postponed because of the patient's serious cardiac medical history and she was referred to us for conservative treatment. She was found to have iliopsoas bursitis with chondrocalcinosis of the knee. Local steroid injections of the two sites abolished her symptoms. We draw attention to the possible pitfalls that the radiographic appearance and one of the multiple clinical presentations of this unrare pathology may represent. Whenever a patient comes walking with crutches, avoids putting weight on his or her leg, and radicular pain is suspected, we advise consideration of other extra spinal causes for the pain. PMID- 9391809 TI - Chondromyxoid fibroma of the ala of the sacrum presenting as a cause of lumbar pain in an adolescent. AB - We report a case of chondromyxoid fibroma of the ala of the sacrum: its presentation, diagnosis, treatment, and resolution. Although this tumor is admittedly rare, our case demonstrates the need for careful evaluation of pack pain in an adolescent. PMID- 9391810 TI - Congenital absence of the pedicles and the neural arch of L2. AB - Congenital pedicle abnormalities are rare. Unilateral aplastic and hypoplastic lumbar pedicles have been reported, but these were usually discovered incidentally and did not need surgical treatment. We present a case of absence of both pedicles and the neural arch of L2, with associated kyphoscoliosis with neurological involvement, that needed a two-stage corrective surgery. An L1-L4 fusion was achieved with relief of the symptoms. PMID- 9391812 TI - Fractures of the calcaneus: open reduction and internal fixation from the medial side a 21-year prospective study. AB - Since 1974, 61 displaced fractures of the calcaneus have been treated by open reduction and internal fixation by a modified medial approach technique. Surgery was performed through a 5-cm incision posterior to the neurovascular bundle. A single threaded pin was passed longitudinally through the tuberosity and into the sustentacular fragment, giving stable fixation. Reduction of the depressed posterior facet fragments was accomplished from the medial side in 77% of cases, occasionally assisted by fluoroscopy. Postoperatively all fractures were immobilized in a cast for 4 weeks. At the end of 4 weeks the pin was removed, and full weight bearing in a walking cast was started and continued for 4 weeks. At 8 weeks after surgery, the walking cast was removed, and the patient began walking in a shoe. These cases were evaluated at a mean follow-up of 4.4 years. There were 49 successful cases (80.3%) and 12 unsuccessful cases (19.7%). A high number of superior results was found in the successful group as shown by the mean score of 94.7 (American Orthopaedic Foot & Ankle Society Scoring System). Time to return to work was a mean of 4.9 months. PMID- 9391813 TI - Radiographic and CT evaluation of tibiofibular syndesmotic diastasis: a cadaver study. AB - Twelve cadaver lower limbs were used for radiographic and CT assessment of the tibiofibular syndesmosis. Plastic spacers were placed in the distal tibiofibular intervals of each specimen in successive 1-mm increments until diastasis could be appreciated on the plain radiographs. All 2- and 3-mm diastases could be noted and clearly identified on CT scans, while the 1-, 2-mm, and half of the 3-mm syndesmotic diastases could not be appreciated with routine radiographs. CT scanning is more sensitive than radiography for detecting the minor degrees of syndesmotic injuries. Therefore, a CT scan can be performed in cases of syndesmotic instability after ankle injuries and for preoperative or postoperative evaluation of the integrity of the distal tibiofibular syndesmosis in cases of doubtful condition of the syndesmosis. PMID- 9391814 TI - Charcot ankle fusion with a retrograde locked intramedullary nail. AB - Twenty patients with severe neuropathic (Charcot) ankle deformities underwent 21 attempted ankle fusions with a retrograde locked intramedullary nail as an alternative to amputation. All had insensate heel pads and had failed at nonoperative methods of accommodative ambulatory bracing. In 11, the talus was either absent, or the deformity was of sufficient magnitude to require talectomy to align the calcaneus under the tibia for plantigrade weightbearing. Ages ranged from 28 to -68 (average 56.3) years. Nineteen were diabetic, 12 being insulin dependent. Their average body weight was 102 kg, with 11 greater than 90 kg at the time of surgery. Eight had chronic large full thickness ulcers overlying, but not involving bone of the medial malleolus, medial midfoot, or proximal fifth metatarsal, at the time of surgery. At a follow-up of 12 to 31 months, 19 achieved bony fusion. In the 10 patients where talectomy was not required, fusion was achieved at an average of 5.3 months without complications. In the patients who required talectomy, six of the patients required eight additional operations to achieve fusion. Three achieved fusion following removal of the nail and prolonged bracing. One opted for ankle disarticulation for chronic persistent infection, rather than attempt reoperation. One died of unrelated causes during the early postoperative period. Retrograde locked intramedullary ankle fusion is a reasonable alternative to amputation in the neuropathic (Charcot) ankle that cannot be controlled with standard bracing techniques. The potential for morbidity requiring reoperation is greatly increased when the deformity is of sufficient magnitude to require talectomy to achieve alignment of the calcaneus in a plantigrade weight-bearing position under the tibia or when there are large open ulcers. PMID- 9391811 TI - Lumbar epidural perineural injection: a new technique. AB - Two controlled studies for a new epidural, perineural, single-shot, selective nerve root injection with a double-needle approach to the anterior epidural space of the lumbar spinal canal are presented. The results were analysed to determine the effectiveness of the new epidural perineural injection technique. The trial comprised two controlled studies on 182 patients. One study compared prospectively randomized results of patients with lumbar radicular syndromes who received epidural perineural injections (n = 47), conventional posterior epidural injections (n = 40) and, as a control group, paravertebral local anaesthetic (n = 46). A second, prospective, double-blind study compared the effect of epidural perineural injections with triamcinolone (n = 24) and pure saline (n = 25). Epidural perineural injections were more effective than conventional posterior epidural injections. Both epidural groups had better results than the paravertebral local injection group. Epidural perineural injections with steroids (10 mg triamcinolone) were more effective than saline alone. A systemic steroid effect was excluded by additional intramuscular steroid injections in the saline group. There were no severe complications or side effects in any of the three groups. The studies concluded that single-shot epidural perineural injection is effective in the treatment of lumbar radicular pain. It is a "one drop only" therapy to the source of pain. PMID- 9391815 TI - Alterations in talar morphology associated with adult flatfoot. AB - To gain a better understanding on the anatomy of the factors contributing to symptomatic flatfoot, we compared the shape of the talus in feet that were flat to that in control tali from feet with a normal arch. Computed tomographic (CT) scans were performed on 9 adult patients with 10 symptomatic flatfoot deformities. CT scans of 10 feet being evaluated for acute trauma not involving the talus were randomly selected as controls. Flatfoot tali tended to be of greater overall length than the control tali, and this difference was not statistically significant. Statistically significant differences were found when comparing ratios of talar length with talar width (P = 0.011), talar length with talar height (P = 0.001) (they were long relative to their height and width), and head length with head width (P = 0.001) for individual tali from the two groups. The tali from the flatfoot group were narrower in width and shorter in height when compared with overall length and had heads that were more elongated in the transverse plane than tali in feet with a normal appearance. CLINICAL CORRELATION: When performing surgical correction of a flatfoot in an adult, appearance of the foot rather than standard radiographic parameters should be used to judge the reduction. The altered shape of the bone may alter the standard radiographic parameters. PMID- 9391816 TI - Subtalar arthrodesis versus flexor digitorum longus tendon transfer for severe flatfoot deformity: an in vitro biomechanical analysis. AB - We defined the mechanical behavior of the foot after an operation for treatment of the flatfoot deformity, subtalar arthrodesis, and compared results with those from flexor digitorum longus tendon transfer. Twelve fresh-frozen human foot specimens were used. Supporting elements were sectioned to create a flatfoot deformity. To simulate midstance phase of gait, loads were applied axially to the plantar foot and to five tendons. Reduction of deformity in metatarsal-talar, calcaneal-talar, and talar-tibial positions was achieved and was significantly greater after subtalar arthrodesis operation than after flexor digitorum longus transfer. PMID- 9391817 TI - Clinical characteristics and outcome in 223 diabetic patients with deep foot infections. AB - Clinical characteristics and outcome in 223 consecutive diabetic patients with deep foot infections are reported. Patients were treated by a multidisciplinary diabetic foot-care team at the University Hospital, Lund, Sweden, and were prospectively followed until healing or death. About 50% of patients lacked clinical signs of infection, such as a body temperature > 37.8 degrees C, a sedimentation rate > 70 mm/hour, and white blood cell count (WBC) > 10 x 10(9)/liter. Eighty-six percent had surgery before healing or death. Thirty-nine percent healed without amputation; 34% healed after a minor and 8% after a major amputation. Sixteen percent were unhealed at death, and 3% were unhealed at the end of the observation period. Of those unhealed at death or follow-up, 4 patients had had a major and 11 a minor amputation. After correction for age and sex, duration of diabetes < 14 years, palpable popliteal pulse, a toe pressure > 45 mmHg, and an ankle pressure > 80 mm Hg, absence of exposed bone and a white blood cell count < 12 x 10(9)/liter were all related to healing without amputation in a logistic regression analysis. We conclude that although only 1 in 10 had a major amputation, nearly all diabetic patients with a deep foot infection needed surgery and more than one third had a minor amputation before healing or death in spite of a well-functioning diabetic foot-care team responsible for all included patients. PMID- 9391818 TI - Reconstruction of the lateral ankle ligaments using an inferior extensor retinaculum flap. AB - The aim of this study was to assess the results of 32 cases of chronic ankle instability. These were treated by ligament shortening and reinforced with an inferior extensor retinaculum flap. All patients complained of persistent functional instability unrelieved with proprioceptive exercises. Results were assessed clinically (pain, instability, recovery of sports activity, mobility) and radiologically (correction of laxity on stress x-rays). This enabled us to draw up a revision score on a scale of 100 points. We obtained a mean score of 86.7 points (45-100 points), and subjective results showed that 88% of the patients were satisfied with the surgery. PMID- 9391819 TI - A kinematic study of ankle joint instability due to rupture of the lateral ligaments. AB - Kinematic analysis of patients who have ruptured the ligaments of the ankle joint was performed to evaluate the function of those ligaments. Ten patients with ruptured lateral collateral ligaments and 10 normal volunteers were examined. Patients' ages ranged from 17 to 29, averaging 20.9 years old. We performed kinematic evaluation by a three-dimensional optical analytic technique using surface markers. According to our results, the ankles with lateral ligament injury abnormally pronated and rotated externally at the time of heel strike and abnormally supinated and rotated internally during the acceleration phase. PMID- 9391820 TI - Isolated dislocation of the middle cuneiform in a farmer: a case report and review of the literature. AB - Isolated dislocations of the middle cuneiform are uncommon. There have been four reported previously. Significant force is required to produce these injuries, and they can have serious neurovascular consequences. In this case report and review of the literature, we present an isolated middle cuneiform dislocation in a 69 year-old farmer with impending skin and soft tissue loss over the dislocated bone. PMID- 9391821 TI - Metatarsal lengthening: case report and review of literature. AB - Shortening of one or more metatarsals may be a cause of metatarsalgia and painful toe deformity. Usually, symptoms are limited and may be successfully addressed with nonoperative treatment. Rarely, operation indicated. This report reviews the surgical techniques, results, and complications. These operations include osteotomy and one-stage distraction with bone grafting, osteotomy and one-stage distraction without bone grafting, osteotomy with gradual distraction and bone grafting, and osteotomy with gradual distraction without bone grafting. PMID- 9391822 TI - Adamantinoma arising in the distal fibula treated with distal fibulectomy: a case report and review of the literature. AB - Adamantinoma is a rare primary bone tumor occurring in the mandible and the long tubular bones. The diaphysis of the tibia is the most common site of extragnathic presentation. Fibular involvement is rare and usually has coexisting tibial involvement. Adamantinoma arising in the distal fibular metaphysis has not been previously reported. This is a case of a teenage boy presenting with a cystic lesion of the distal fibula, initially diagnosed and treated as a unicameral bone cyst. Aggressive behavior ultimately led to a diagnosis of adamantinoma. He was treated with distal fibulectomy without surgical reconstruction with good functional outcome. PMID- 9391823 TI - Linkage disequilibrium measures for fine-scale mapping: a comparison. AB - We investigate properties of simple linkage disequilibrium mapping for five measures in the presence of mutation at the marker and/or the disease locus and of initial incomplete linkage disequilibrium. In contrast to the stimulation approach that Devlin and Risch used, we calculate the expected values of various linkage disequilibrium measures under different assumptions based on a framework for linkage disequilibrium mapping. These expected values clearly demonstrate the expected performance of these measures. We find that the impact of marker mutation on their performance depends on the magnitude of the mutation relative to the proximity of the marker (i.e. recombination fraction between the marker and the disease locus). In the presence of recurrent mutation at the marker and/or disease locus, the performance of all measures, including the robust one, depends on the marker allele frequency. The initial incomplete linkage disequilibrium could render all measures useless. These expected values also show clearly why in Devlin and Risch's simulation some measures performed very badly under certain circumstances. PMID- 9391824 TI - Estimating the age of mutant disease alleles based on linkage disequilibrium. AB - With more and more disease genes being mapped and/or cloned, there is a growing interest in dating the age of underlying mutations. The knowledge of the age of mutation is important to finely map disease genes by linkage disequilibrium mapping. It would also help us understand the origin, evolution, and dispersion of the mutant disease genes. Despite increasing interests in dating disease mutations, the development of appropriate statistical methods is largely fragmentary, and there is a lack of systematic treatment of the topic. We propose two classes of methods for estimating the age of mutant allele at the disease locus based on linked marker data. Our methods can not handle only single-locus marker data, but also multi-locus marker data as well. Moreover, our methods can be used even when the location of the disease locus is unknown, and/or when there are mutations at the marker or disease locus. We show that some previous results are special cases of our methods. We also derive a recursive equation previously obtained by Serre et al. [Hum Genet 1990;84:449-454] and provide an explicit solution to the equation. To illustrate our methods, we applied them to two groups of data sets, one is cystic fibrosis data collected from several European populations, and the other is data on several genetic diseases (diastrophic dysplasia, progressive myoclonus epilepsy, congenital chloride diarrhea, and Batten disease) all collected from the Finnish population. The former data set allows us to trace the origin and dispersion of the most common mutation for cystic fibrosis. The latter provides an opportunity to examine whether all mutations for these diseases have the same age. PMID- 9391825 TI - AvaI polymorphism in the human transferrin gene. AB - A guanine-adenine substitution was observed in exon 5 of the human transferrin (TF) gene. The nucleotide change led to an AvaI digestion site. Analysis of the segregation of the AvaI polymorphism and serum TF phenotypes indicated that an intragenic recombination occurred between the AvaI polymorphic site and the mutation site in the TF gene which determines the two common TF alleles, TF*C1 and TF*C2. PMID- 9391826 TI - A transmission disequilibrium test for quantitative trait loci. AB - The transmission disequilibrium test uses association between marker alleles and dichotomous traits for precise genetic mapping while avoiding confounding due to population admixture. Here, the methodology is generalized from dichotomous traits to quantitative traits. The generalization is computationally straightforward and may be used with multiple alleles and with siblings. Parametric assumptions on the distribution of the quantitative traits are not needed. Environmental and demographic covariates may be incorporated into the analysis. The results of simulation studies that provide information about the power of the approach are reported. PMID- 9391827 TI - Feasibility of collecting disease reports from relatives for genetic epidemiologic investigations. AB - Self-reports of disease from relatives are generally believed to be more detailed than those received from a family informant, although differential participation may exist among the relatives who provide information. To investigate the potential for differential participation, we requested permission to contact relatives of mothers (informants) who had provided family history information for a population-based case-control study of orofacial clefts. Birth defect and cancer self-reports were received from 345 (65.6%) case and 380 (68.8%) control relatives. Participants and nonparticipants differed little by type (maternal or paternal) or degree of relationship. Informants, however, were more likely to permit contact with relatives who were maternal, first-degree and female. Relatives appeared willing to provide self-reports, although the potential for differential selection introduced by informants should be considered. PMID- 9391828 TI - Structural and functional adaptations of skeletal muscle to weightlessness. AB - This brief review examines the effects of weightlessness on the structure and function of human and rat skeletal muscle. Data collected from spaceflights or Earth-based models suggest that a rapid atrophy (5-8 days) occurs in lower limb muscles associated with impairments in muscle strength, physical work capacity and locomotor coordination. The reduction in muscle cross-sectional area cannot entirely explain the loss of strength in postural muscles suggesting a reduced neural drive. Muscle atrophy is accompanied by a general shift in the contractile and enzymatic profiles of a slow-twitch oxidative muscle toward that of a fast twitch glycolytic muscle. In humans, limited structural and functional data collected in astronauts after short periods of microgravity are qualitatively similar to those observed in rats after real or simulated microgravity suggesting that animal models are relevant to muscle research in space. A preventive prescription i.e. exercise or pharmacological treatment, cannot be proposed until future research has better defined the basic mechanisms of muscle plasticity during long duration spaceflights. PMID- 9391829 TI - Molecular events underlying skeletal muscle atrophy and the development of effective countermeasures. AB - Skeletal muscle adapts to loading; atrophying when exposed to unloading on Earth or in spaceflight. Significant atrophy (decreases in muscle fiber cross-section of 11-24%) in humans has been noted after only 5 days in space. Since muscle strength is determined both by muscle cross-section and synchronization of motor unit recruitment, a loss in muscle size weakens astronauts, which would increase risks to their safety if an emergency required maximal muscle force. Numerous countermeasures have been tested to prevent atrophy. Resistant exercise together with growth hormone and IGF-I are effective countermeasures to unloading as most atrophy is prevented in animal models. The loss of muscle protein is due to an early decrease in protein synthesis rate and a later increase in protein degradation. The initial decrease in protein synthesis is a result of decreased protein translation, caused by a prolongation in the elongation rate. A decrease in HSP70 by a sight increase in ATP may be the factors prolonging elongation rate. Increases in the activities of proteolytic enzymes and in ubiquitin contribute to the increased protein degradation rate in unloaded muscle. Numerous mRNA concentrations have been shown to be altered in unloaded muscles. Decreases in mRNAs for contractile proteins usually occur after the initial fall in protein synthesis rates. Much additional research is needed to determine the mechanism by which muscle senses the absence of gravity with an adaptive atrophy. The development of effective countermeasures to unloading atrophy will require more research. PMID- 9391830 TI - The contribution of NMR, NIRS and their combination to the functional assessment of human muscle. AB - In the last decade the study of the human muscle mechanics and energetics in physiology and pathology underwent a radical change. Indeed, the use of biopsy is being progressively accompanied by non-invasive techniques which allow a more integrative assessment of muscle function in vivo. Magnetic resonance imaging (MRI) provides a better insight into muscle structure down to the fascicular level, proving to be an essential source of information, particularly for the study of biomechanics. Magnetic resonance spectroscopy (MRS), besides the study of muscle anaerobic (high energy phosphate compounds by 31P and 1H) and aerobic metabolism as well as of metabolic control, makes it possible to follow the time course of glycogen concentration (13C) as a function of exercise intensity and duration and of its recovery both in healthy trained and untrained subjects and in diabetic patients. Near-infrared spectroscopy (NIRS) allows to assess muscle oxidative metabolism, providing changes of total, deoxy- and oxy-hemoglobin in the resting and contracting muscle. The concomitant use of the above techniques is expected to provide a synergic functional picture of the human muscle that is complementary to the structural and ultrastructural microscopic approach. PMID- 9391831 TI - Recommendations for muscle research in space. PMID- 9391832 TI - Muscle atrophy during long duration bed rest. PMID- 9391833 TI - Changes in mechanical properties of human muscle as a result of spaceflight. PMID- 9391834 TI - The effect of prolonged bed rest on maximal instantaneous muscle power and its determinants. PMID- 9391835 TI - Changes in electrically evoked skeletal muscle contractions during 17-day spaceflight and bed rest. PMID- 9391836 TI - Muscle fiber atrophy and degeneration induced by experimental immobility and hindlimb suspension. PMID- 9391837 TI - Role of gravitational loading on the development of soleus muscle in rats. PMID- 9391838 TI - Interactive effects of loading and thyroid states on skeletal isomyosins. PMID- 9391839 TI - Postnatal muscle development in unloading conditions. PMID- 9391840 TI - Molecular biology of human muscle adaptation. PMID- 9391841 TI - Functional and biochemical adaptations to low-frequency stimulation: possible applications to microgravity. PMID- 9391842 TI - Integrated human muscle bioenergetic studies by magnetic resonance methods. PMID- 9391843 TI - Muscle energetics by 31P-NMRS: recent developments and possible applications in space research. PMID- 9391844 TI - Changes in calf muscle performance, energy metabolism, and muscle volume caused by long-term stay on space station MIR. PMID- 9391845 TI - MR-spectroscopy (MRS) of different nuclei applied to human muscle: additional information obtained by 1H-MRS. PMID- 9391846 TI - Metabolic studies of human skeletal muscle by near infrared spectroscopy: possible applications in space research. PMID- 9391847 TI - Eccentric exercise and muscle damage. PMID- 9391848 TI - Muscle research in space--increased muscle susceptibility to exercise-induced damage after a prolonged bedrest. PMID- 9391849 TI - Mechanism of fatigue in small muscle groups. PMID- 9391850 TI - Resistance training in space. PMID- 9391851 TI - Cycling in space to simulate gravity. PMID- 9391852 TI - Stretch-shortening-cycle in microgravity, spinal proprioceptive and tendomuscular elastic energy release. PMID- 9391853 TI - Activation of muscles and microgravity. PMID- 9391854 TI - Continuous monitoring of spine geometry: a new approach to study back pain in space. PMID- 9391855 TI - Treatment of endstage heart disease with mechanical circulatory assistance. AB - A great number of patients suffer and die from the sequelae of acute and chronic heart failure each year. Although advances in medical and surgical therapy have benefited many of these patients, the majority suffer from disease refractory to any definitive therapy. For these patients, cardiac transplantation is the only remaining hope. Unfortunately, because of the increasing demand for donor organs in the face of a fixed and limited supply, this option is only available to a small percentage of these patients. Even in patients accepted for transplantation, a significant waiting list mortality has been observed. A variety of ventricular assist devices (VAD) have been developed since the first successful case of mechanical cardiac assistance over 30 years ago. These devices differ in basic mechanical function, method of insertion, and degree of implantability, and thus have different indications and potential applications. While the intra-aortic balloon pump and centrifugal pumps are effective short term support modalities, extracorporeal and implantable pulsatile devices have been used successfully for long-term support of patients with reversible and non reversible cardiac failure. These pumps have most commonly been utilized as bridges to transplantation, but increasing clinical experience has supported the notion of long-term mechanical assistance as a definitive therapy for endstage heart disease. While complications, particularly infection and thromboembolism, pose significant challenges and long-term device reliability remains to be fully determined, available implantable devices seem capable of providing effective long-term support. As data is obtained from currently ongoing trials comparing VAD support to medical therapy for endstage heart failure, ethical and economic issues will assume increasing importance. PMID- 9391856 TI - Arterial wall neovascularization--potential role in atherosclerosis and restenosis. AB - Neointimal formation and arterial wall remodeling are pivotal causes of luminal narrowing in atherogenesis and restenosis. Arterial remodeling refers to a series of dynamic structural changes that arteries may undergo in response to various stimuli, including changes in blood flow and pressure, and acute injury. The biological mechanisms involved in arterial remodeling are poorly understood and are currently a main target for research. We have recently focused on the role of the arterial wall microcirculation (ie, vasa vasorum) in arterial remodeling after injury. In the past, a correlation between arterial wall neovascularization and the accumulation of arterial plaque has been documented; however, the dynamic role of these microvessels in arterial repair and luminal narrowing has not been examined. The type of arterial injury, the nature of the lesion that develops, and the arterial compartment in which angiogenesis occurs may determine the role of the vasa vasorum in arterial narrowing. In this review, we highlight the data that link arterial wall neovascularization with lesion formation and the process of arterial remodeling. PMID- 9391857 TI - An enhanced method for measuring cardiac output using Doppler color flow echocardiography. AB - An enhanced method for determining cardiac output using Doppler color flow imaging techniques to measure mitral orifice diameter was developed and validated in an experimental model and in clinical patients. In an in vitro circuit model, color jet width correlated well with actual orifice dimension from 12 to 24 mm (r = 0.99). In the clinical application, mitral valve area was calculated as a X b X pi/4 where a and b represent the width of the color flow stream in the mitral orifice just distal to the annulus in apical long-axis (short-diameter) and 4 chamber (90 degrees rotated, long-diameter) views, respectively. Cardiac output was then computed as the product of mitral valve area and time-velocity integral of transmitral flow from the same site. Cardiac output was also measured by thermodilution and conventional echocardiographic methods using diameters and time-velocity integrals from the left ventricular outflow tract. In 30 patients with nonvalvular heart disease, cardiac output measured by thermodilution ranged from 3.40 to 8.40 L/min. Cardiac output was determined in 28 of 30 patients (93%) by the Doppler color flow imaging technique; it ranged from 3.00 to 8.36 L/min and correlated well with thermodilution: y = 0.90x + 0.63, r = 0.91. Cardiac output was determined in 24 of 30 patients by the conventional left ventricular outflow method (80%). The cardiac output measured by the conventional method correlated less closely with thermodilution (r = 0.84), although there was no statistical difference in correlation coefficiencies between the 2 methods. These results indicate that the Doppler color flow imaging technique can be used to enhance the determination of cardiac output by echocardiography, particularly when the conventional method has resulted in technically inadequate recordings. PMID- 9391858 TI - Mitral insufficiency as a complication of acute myocardial infarction and left ventricular remodeling. AB - The presence of mitral insufficiency after acute myocardial infarction (AMI) often leads to hemodynamic impairment and heart failure. This study was designed to examine the relationship between mitral regurgitation (MR), an indicator of mitral insufficiency, and the course of recovery from AMI. We evaluated the course of MR after AMI in 223 patients by color Doppler echocardiography. MR was detected in 21% (47/223) of patients at the onset of AMI, and developed in 18% (40/223) of patients during follow-up. Patients were grouped according to the course of MR as well as the success of acute recanalization therapy. No correlation was observed between the presence or course of MR and the site of infarction. The incidence of successful recanalization was higher in patients with MR that improved during follow-up than in patients with MR that was unchanged or that worsened during follow-up. Although no significant differences in hemodynamic variables were noted among the groups at admission, the group with unsuccessful recanalization and unimproved MR (BS-) showed a significantly greater left ventricular end-diastolic diameter (LVDd), left ventricular end systolic diameter, and left ventricular end-diastolic volume (LVEDV) as well as a lower left ventricular ejection fraction than patients with successful recanalization and no MR (CS+) during the convalescent period. The extent of change in LVDd and LVEDV between admission and convalescence was significantly greater in the BS(-) group than in the CS(+) group. The results suggest that successful recanalization after AMI reduces the incidence of MR. Acute recanalization therapy after AMI may prevent left ventricular remodeling, resulting in a secondary improvement of MR. PMID- 9391859 TI - Induction of ST-segment elevation by regional myocardial stretch in normal canine hearts in vivo. AB - The purpose of this study was to evaluate ST-segment elevation induced by regional myocardial stretch without myocardial ischemia in canine hearts. A strain gauge arch (TH-601T) was sutured to the left ventricular epicardium, parallel to the short axis, to shorten the end-diastolic length of the myocardium beneath the arch (stretch zone; SZ) and to produce regional myocardial stretch in each of 6 dogs. An increase in preload caused by altering the height of a saline filled reservoir affected prolongation or shortening of the myocardium both in the SZ and outside the arch (normal zone; NZ) to increase myocardial stretch. An epicardial electrocardiogram was recorded in both the SZ and the NZ. After suture of the strain gauge arch, the ST segment was elevated in the SZ. An increase in preload augmented stretch during systole in the SZ, resulting in additional ST segment elevation. These results suggest that regional myocardial stretch itself plays an important role in ST-segment elevation. PMID- 9391860 TI - Effects of alpha 1-adrenoreceptor subtype blockade on ischemia-reperfusion injury. AB - To clarify the roles of subclasses of alpha 1-adrenoreceptors in ischemic reperfused myocardium, we compared the effect of the nonselective alpha 1-blocker bunazosin with that of the alpha 1A-blocker WB4101 and the alpha 1B-blocker chlorethylclonidine (CEC) in isolated rat hearts. After 30 min of preperfusion, Langendorff-perfused hearts were subjected to 25 min of global ischemia followed by 30 min of reperfusion. Hearts were randomly divided into 4 groups, with one of the following substances being added to the perfusate: buffer alone (control), 10(-6) mol/L bunazosin, 10(-7) mol/L WB4101, or 10(-7) mol/L CEC. Bunazosin had a negative inotropic effect and preserved the postischemic ATP content, reduced the postischemic increase in intracellular Na+ content and then enhanced postreperfusion recovery of creatine phosphate. Bunazosin also reduced myocardial 45Ca2+ uptake during reperfusion (control 5.2 vs bunazosin 2.5 mumol/g dry weight of tissue (dwt), p < 0.01). However, the recovery of left ventricular developed pressure (DP) was not improved when bunazosin was added to the perfusate during reperfusion. WB4101 had neither a negative inotropic nor an energy-sparing effect, but it improved the recovery of DP (control 43% vs WB4101 56% of preischemic value, p < 0.05) with no reduction in myocardial 45Ca2+ uptake. CEC had a negative inotropic and energy-sparing effect and then reduced myocardial 45Ca2+ uptake (CEC 3.1 mumol/g dwt, p < 0.05), but it did not improve the recovery of DP. These results suggest that the preischemic administration of an alpha 1B-adrenoreceptor subtype blocker protected ischemic-reperfused myocardium via reduction of Ca2+ overload, whereas the selective blockade of the alpha 1A adrenoreceptor subtype reduced myocardial damage via mechanism(s) other than Ca2+ metabolism. PMID- 9391861 TI - The delayed recovery of impaired endothelium-dependent vasodilatory response after hemodynamic improvement in dogs with congestive heart failure. AB - We investigated whether impaired endothelium-dependent vasodilatory response recovers as heart failure improves. The femoral blood flow responses to intra arterial administration of nitroglycerin (NTG) and acetylcholine (ACh) were examined in dogs with 2-week pacing-induced chronic congestive heart failure (congestive heart failure group; CHF, n = 12). Thereafter, pacing was stopped and hemodynamic changes and femoral blood flow responses were re-examined either 1 week (recover 1 week group; Re 1W, n = 6) or 4 weeks (recover 4 weeks group; Re 4W, n = 6) after the cessation of pacing. Another group in which a pacemaker was implanted without pacing served as the control group (n = 8). In CHF, heart rate and pulmonary artery pressure increased, and echocardiography revealed increased left ventricular diastolic dimension and reduced percent fractional shortening compared with those in the control group. In Re 1W, all hemodynamic parameters returned to the basal levels and did not differ from those in the control group. Although there was no significant difference in the blood flow responses to NTG among the 4 groups, the responses to ACh in CHF were significantly reduced compared with those in the control group. Despite the recovered hemodynamics, femoral blood flow responses to ACh were still reduced in Re 1W, but they returned to the levels of the control group in Re 4W. Thus, vascular endothelial dysfunction recovers along with improvement in CHF, however, the recovery of endothelial function is delayed in comparison with improvement in cardiac function. PMID- 9391862 TI - A case of primary malignant fibrous histiocytoma of the heart with a left-to right atrial shunt. AB - A previously healthy 64-year-old woman attended our hospital with chest pain, facial edema, and general fatigue. A chest radiograph revealed cardiomegaly, small bilateral pleural effusions, and hilar congestion--findings that improved after early therapy with furosemide and methyldigoxin. A chest radiograph recorded 7 years earlier had revealed no dilation of cardiac shadow. There were no findings suggesting atrial septal defect (ASD) or valvular heart disease. Echocardiography revealed a tumor-like mass adhering to the posterior wall of the left atrium. Color-flow Doppler echocardiography revealed a left-to-right shunt at the atrial level. The Qp/Qs ratio as measured by cardiac catheterization was 2.0. Coronary angiography revealed abnormal dilated arteries from the atrioventricular nodal branch and several feeding arteries from the left circumflex branch. We hypothesized that the left-to-right shunt could be due to the tumor, which extended to the rim of the patent foramen ovale, or to the very small, previously unrecognized, ASD. This patient died 6 months after her first admission and an autopsy was performed. Light microscopic examination of the tumor revealed spindle-shaped fibroblast-like cells arranged in a storiform or fascicular pattern. The immunohistochemical findings were consistent with malignant fibrous histiocytoma (MFH). In the literature, left-to-right shunt at the atrial level has not been reported in patients with cardiac MFH. PMID- 9391863 TI - The coexistence of abdominal aortic aneurysm and advanced gastric cancer associated with recurrent angina after coronary artery bypass grafting. AB - A 76-year-old man with abdominal aortic aneurysm (AAA) and concomitant gastric cancer, who had undergone coronary artery bypass grafting (CABG), presented with recurrent exertional angina. Both lesions, the AAA and advanced gastric cancer, exhibited an absolute indication for urgent surgery. Coronary revascularization with percutaneous transluminal coronary angioplasty (PTCA) was carried out successfully before abdominal surgery. A one-stage abdominal operation was performed safely. The need for coronary revascularization complicates the treatment strategy for these patients with associated coronary artery disease. PTCA is the best option, especially if the patient presents with recurrent angina after prior CABG. PMID- 9391864 TI - Ventricular septal defect secondary to non-penetrating chest trauma. AB - A 52-year-old man acquired a ventricular septal defect following non-penetrating chest trauma. Four days after the traumatic accident, he showed signs of congestive heart failure. Imaging techniques using echocardiography and left ventriculography were helpful in diagnosing the condition. Surgical repair by patch closure of the ventricular septal defect was accomplished 7 days after the traumatic accident. PMID- 9391865 TI - A case of acute myocardial infarction due to primary coronary dissection. AB - A case of acute myocardial infarction associated with primary coronary dissection was followed up angiographically. A 46-year-old woman complained of chest oppression. Electrocardiogram on admission showed ST-segment elevation in V1-5. Urgent coronary angiography was performed under a diagnosis of acute anterior myocardial infarction, and showed a significant stenosis with multiple filling defects in segments 7-8 (99% with severe delay) in the left anterior descending artery. There was no organic lesion in the right coronary artery. Intracoronary thrombolytic therapy was unsuccessful, and thereafter she was treated with aspirin, warfarin and isosorbide dinitrate. Coronary angiography performed 1 month later revealed a long dissection with double lumens in segments 7-8. The septal branches emerged from the smaller lumen. Two months later, the 2 lumens were almost equal in size. These findings indicated that coronary dissection produced a false lumen with an entry in segment 7 and a reentry in segment 8, and that the false lumen was responsible for the greater flow. Four months later, the flow in the true lumen had improved remarkably while that in the false lumen had almost disappeared. She remained in stable condition during the follow-up period of 4 months. PMID- 9391866 TI - Successful thromboendarterectomy for chronic pulmonary embolism in a patient with systemic lupus erythematosus and antiphospholipid syndrome. AB - Chronic pulmonary thromboembolism is known to be associated with poor prognosis with conservative medical treatment. Pulmonary thromboendarterectomy for chronic pulmonary thromboembolic disease is a potentially lifesaving procedure that prevents right-sided cardiac failure as a result of secondary pulmonary hypertension caused by pulmonary thromboembolism. We report a rare case of systemic lupus erythematosus with antiphospholipid syndrome in a patient who presented with pulmonary hypertension secondary to chronic proximal multiple pulmonary thromboembolism. To our knowledge, this is the first case report of chronic pulmonary thromboembolism complicated by systemic lupus erythematosus with antiphospholipid syndrome. Thromboendarterectomy was performed with satisfactory results. PMID- 9391867 TI - The viscoelastic behavior of the non-degenerate human lumbar nucleus pulposus in shear. AB - The viscoelastic behavior of the nucleus pulposus was determined in shear under transient and dynamic conditions and was modeled using a linear viscoelastic model with a variable amplitude relaxation spectrum. During stress-relaxation tests, the shear stress of the nucleus pulposus relaxed nearly to zero indicative of the fluid nature of the tissue. Under dynamic conditions, however, the nucleus pulposus exhibited predominantly 'solid-like' behavior with values for dynamic modulus (magnitude of G*) ranging from 7 to 20 kPa and loss angle (delta) ranging from 23 to 30 degrees over the range of angular frequencies tested (1-100 rad s 1). This frequency-sensitive viscoelastic behavior is likely to be related to the highly polydisperse populations of nucleus pulposus molecular constituents. The stress-relaxation behavior, which was not linear on a semi-log plot (in the range t1 << t << t2), required a variable amplitude relaxation spectrum capable of describing this frequency sensitive behavior. The stress-relaxation behavior was well described by this linear viscoelastic model with variable amplitude relaxation spectrum; however, the dynamic moduli were underpredicted by the model which may be related to non-linearities in the material behavior. PMID- 9391868 TI - Dependence of cruciate-ligament loading on muscle forces and external load. AB - A sagittal-plane model of the knee is used to predict and explain the relationships between the forces developed by the muscles, the external loads applied to the leg, and the forces induced in the cruciate ligaments during isometric exercises. The geometry of the model bones is adapted from cadaver data. Eleven elastic elements describe the geometric and mechanical properties of the cruciate ligaments, the collateral ligaments, and the posterior capsule. The model is actuated by 11 musculotendinous units, each unit represented as a three element muscle in series with tendon. For isolated contractions of the quadriceps, ACL force increases as quadriceps force increases for all flexion angles between 0 and 80 degrees; the ACL is unloaded at flexion angles greater than 80 degrees. When quadriceps force is held constant, ACL force decreases monotonically as knee-flexion angle increases. The relationship between ACL force, quadriceps force, and knee-flexion angle is explained by the geometry of the knee-extensor mechanism and by the changing orientation of the ACL in the sagittal plane. For isolated contractions of the hamstrings, PCL force increases as hamstrings force increases for all flexion angles greater than 10 degrees; the PCL is unloaded at flexion angles less than 10 degrees. When hamstrings force is held constant, PCL force increases monotonically with increasing knee flexion. The relationship between PCL force, hamstrings force, and knee-flexion angle is explained by the geometry of the hamstrings and by the changing orientation of the PCL in the sagittal plane. At nearly all knee-flexion angles, hamstrings co contraction is an effective means of reducing ACL force. Hamstrings co contraction cannot protect the ACL near full extension of the knee because these muscles meet the tibia at small angles near full extension, and so cannot apply a sufficiently large posterior shear force to the leg. Moving the restraining force closer to the knee-flexion axis decreases ACL force; varying the orientation of the restraining force has only a small effect on cruciate-ligament loading. PMID- 9391869 TI - Forces of individual cat ankle extensor muscles during locomotion predicted using static optimization. AB - In order to test the principles of the control of synergistic muscles that were proposed in the literature, forces of cat soleus (SO), gastrocnemius (GA), and plantaris (PL) measured during locomotion were compared with the corresponding forces predicted using different optimization criteria. Forces of cat SO, GA, and PL, and the corresponding cat kinematics were recorded simultaneously using E shaped force transducers and high-speed video, respectively. Measurements were obtained from three cats walking and trotting on a treadmill at five nominal speeds ranging from 0.4 to 1.8 m s-1. Muscle forces were predicted using static optimization and a musculoskeletal model of the cat hindlimb consisting of three segments (foot, shank, and thigh) and three muscles (SO, GA, and PL). Six optimization criteria which had been proposed in the literature were tested. Linear criteria based on the principles of minimum muscle force and stress predicted forces during the stance phase with an average normalized error of 59 322%. Three other criteria--minimization of the sum of the relative muscle forces squared, minimization of the sum of the muscle stresses cubed, and minimization of the upper bound for all of the muscle stresses-showed a better performance: (i) the average error was 43-119% and (ii) the correlation coefficient calculated between the predicted and actual forces exceeded 0.9 for all three muscles. A criterion that was based on the principle of minimum fatigue and accounted for the percentage of slow-twitch fibers in the muscles, had the lowest average error (26-52%). The high correlation (0.97-0.99) between the measured forces and forces predicted by using the minimum fatigue criterion suggested that force sharing among SO, GA, and PL during cat locomotion may be the same for a given set of joint moments and muscle moment arms. It was concluded that static optimization with the appropriate criterion can predict muscle forces adequately for specific movement conditions. PMID- 9391870 TI - Force response of the fingertip pulp to repeated compression--effects of loading rate, loading angle and anthropometry. AB - Repeated loading of the fingertips has been postulated to contribute to tendon and nerve disorders at the wrist during activities associated with prolonged fingertip loading such as typing. To fully understand the pathomechanics of these soft tissue disorders, the role of the fingertip pulp in attenuating the applied dynamic forces must be known. An experiment was conducted to characterize the response of the in vivo fingertip pulp under repeated, dynamic, compressive loadings, to identify factors that influence pulp dynamics, and to better understand the force modulation by the pulp. Twenty subjects tapped repeatedly on a flat plate with their left index finger, while the contact force and pulp displacement were measured simultaneously. Tapping trials were conducted at three fingertip contact angles from the horizontal plane (0 degree, 45 degrees, and 90 degrees) and five tapping rates (0.25, 0.5, 1, 2, and 3 Hz). The fingertip pulp responds as a viscoelastic material, exhibiting rate-dependence, hysteresis, and a nonlinear force-displacement relationship. The pulp was relatively compliant at forces less than 1 N, but stiffened rapidly with displacement at higher forces for all loading conditions. This suggests that high-frequency forces of a small magnitude (< 1 N) are attenuated by the nonlinearly stiffening pulp while these forces of larger magnitude are transmitted to the bone. Pulp response was significantly influenced by the angle of loading. Fingertip dimensions, gender, and subject age had little to no influence on pulp parameters. PMID- 9391871 TI - Optimization of cardiac fiber orientation for homogeneous fiber strain at beginning of ejection. AB - Mathematical models of left ventricular (LV) wall mechanics show that fiber stress depends heavily on the choice of muscle fiber orientation in the wall. This finding brought us to the hypothesis that fiber orientation may be such that mechanical load in the wall is homogeneous. Aim of this study was to use the hypothesis to compute a distribution of fiber orientation within the wall. In a finite element model of LV wall mechanics, fiber stresses and strains were calculated at beginning of ejection (BE). Local fiber orientation was quantified by helix (HA) and transverse (TA) fiber angles using a coordinate system with local r-, c-, and l-directions perpendicular to the wall, along the circumference and along the meridian, respectively. The angle between the c-direction and the projection of the fiber direction on the cl-plane (HA) varied linearly with transmural position in the wall. The angle between the c-direction and the projection of the fiber direction on the cr-plane (TA) was zero at the epicardial and endocardial surfaces. Midwall TA increased with distance from the equator. Fiber orientation was optimized so that fiber strains at BE were as homogeneous as possible. By optimization with TA = 0 degree, HA was found to vary from 81.0 degrees at the endocardium to -35.8 degrees at the epicardium. Inclusion of TA in the optimization changed these angles to respectively 90.1 degrees and -48.2 degrees while maximum TA was 15.3 degrees. Then the standard deviation of fiber strain (epsilon f) at BE decreased from +/- 12.5% of mean epsilon f to +/- 9.5%. The root mean square (RMS) difference between computed HA and experimental data reported in literature was 15.0 degrees compared to an RMS difference of 11.6 degrees for a linear regression line through the latter data. PMID- 9391872 TI - The effect of pedaling rate on coordination in cycling. AB - To further understand lower extremity neuromuscular coordination in cycling, the objectives of this study were to examine the effect of pedaling rate on coordination strategies and interpret any apparent changes. These objectives were achieved by collecting electromyography (EMG) data of eight lower extremity muscles and crank angle data from ten subjects at 250 W across pedaling rates ranging from 45 to 120 RPM. To examine the effect of pedaling rate on coordination, EMG burst onset and offset and integrated EMG (iEMG) were computed. In addition, a phase-controlled functional group (PCFG) analysis was performed to interpret observed changes in the EMG patterns in the context of muscle function. Results showed that the EMG onset and offset systematically advanced as pedaling rate increased except for the soleus which shifted later in the crank cycle. The iEMG results revealed that muscles responded differently to increased pedaling rate. The gastrocnemius, hamstring muscles and vastus medialis systematically increased muscle activity as pedaling rate increased. The gluteus maximus and soleus had significant quadratic trends with minimum values at 90 RPM, while the tibialis anterior and rectus femoris showed no significant association with pedaling rate. The PCFG analysis showed that the primary function of each lower extremity muscle remained the same at all pedaling rates. The PCFG analysis, which accounts for muscle activation dynamics, revealed that the earlier onset of muscle excitation produced muscle activity in the same region of the crank cycle. Also, while most of the muscles were excited for a single functional phase, the soleus and rectus femoris were excited during two functional phases. The soleus was classified as an extensor-bottom transition muscle, while the rectus femoris was classified as a top transition-extensor muscle. Further, the relative emphasis of each function appeared to shift as pedaling rate was increased, although each muscle remained bifunctional. PMID- 9391873 TI - Microhardness anisotropy of lamellar bone. AB - The Knoop microhardness test has been utilised to observe in-plane microhardness anisotropy of rat tibiae. The elongated rhombohedral geometry of the Knoop indenter enables the Knoop microhardness (HK) to be calculated for a given indenter orientation. Two indenter orientations were used: the major axis of the indenter was aligned along the length of, and across the mid-sagittal section. The statistical analysis demonstrated that the variation in HK was primarily due to the orientation of the Knoop indenter (p < 0.001). HK was consistently greater when the indenter was aligned with the major diagonal radial on the mid-sagittal section. PMID- 9391874 TI - A three-dimensional finite analysis of adaptive remodelling in the proximal femur. AB - A finite element analysis of adaptive bone remodelling in the proximal femur is presented. The use of a three-dimensional model permits a realistic representation of femur geometry, and also allows the possibility of examining the effects of fully three-dimensional loading situations. The long-term pattern of remodelling shows a realistic evolution of density distribution, with a tendency towards a steady state, though the simplified load cases used to model gait are not sufficient to predict the formation of the cortical shell. PMID- 9391875 TI - An improved method for finite element mesh generation of geometrically complex structures with application to the skullbase. AB - An automated method has been developed to generate finite element meshes of geometrically complex structures from CT images using solely hexahedral elements. This technique improves upon previous voxel-based mesh reconstruction approaches by smoothing the irregular boundaries at model surfaces and material interfaces. Over a range of mesh densities, RMS error in surface Von Mises stress was higher in the unsmoothed circular ring models (0.11-0.24 MPa) than in the smoothed models (0.080-0.15 MPa) at each mesh density. The element-to-element oscillation in surface element stress, as measured by the average second spatial derivative of Von Mises stress along the outer surface of the ring, was higher in the unsmoothed models (11.5-15.0 kPa mm-2) than in the smoothed models (4.0-6.8 kPa mm-2). Similarly, in a human skullbase model, the element-to-element oscillation in surface Von Mises stress was higher in the unsmoothed model (5.52 kPa mm-2) than in the smoothed model (1.83 kPa mm-2). Using this technique, finite element models of complex geometries can be rapidly reconstructed which produce less error at the surface than voxel-based models with discontinuous surfaces. PMID- 9391876 TI - A three-component force vector cell for in vitro quantification of the force exerted by the papillary muscle on the left ventricular wall. AB - Recent clinical studies indicate that functional mitral regurgitation, which is a common complication in patients who suffer from ischemic heart disease, is related to an increase in the tethering forces acting on the mitral valve leaflets. Alterations in the valvular assembly, displacement of the papillary muscles or dilatation of the mitral valve annulus can disrupt the normal force balance on the mitral leaflets and result in an abnormal coaptation geometry with incomplete mitral leaflet closure. The force balance imposed on the mitral leaflets is created by the coapting forces generated by the transmitral pressure difference and the tethering forces at the leaflet attachments. A unique force vector cell capable of accurately measuring the three-component force vector applied by the papillary muscle on the left-ventricular wall was designed and manufactured to permit quantification of the alteration in the force balance acting on the mitral leaflets, and to allow for the study of the influence of papillary muscle displacement on mitral regurgitation. PMID- 9391877 TI - A mechanical device for studying mechanical properties of human muscles in vivo. AB - A mechanical device, primarily devoted to biomechanical studies of human calf muscles during microgravity experiments is presented. It allows investigation of both contractile and visco-elastic properties of musculo-articular systems using, respectively, isokinetic movements, quick-release tests and sinusoidal perturbations. This device is a specifically designed ergometer associated to an experimental protocol designed for pre- and post-flight tests. The protocol was evaluated on 22 healthy subjects and typical results are briefly presented. Preliminary results are discussed in terms of agreement with currently available data and a detailed evaluation of test-retest measurements is provided for quick release experiments. Complementary investigations are suggested and potential fields of research are indicated. PMID- 9391878 TI - Clustering of Y chromosome deletions in subinterval E of interval 6 supports the existence of an oligozoospermia critical region outside the DAZ gene. AB - Y chromosome molecular analysis was performed using the STS-PCR technique in 50 patients with oligozoospermia. Microdeletions of interval 6 of the Y chromosome were detected in seven patients, in six of whom subinterval E was affected. All patients retained the RBM1 and DAZ genes, while in one deletion involved the SPGY gene. The size of the deletion was not apparently related to the severity of the disease. These results suggest the presence of an oligozoospermia critical region on the Y chromosome within subinterval E of interval 6. PMID- 9391879 TI - Low frequency of BRCA1 germline mutations in 45 German breast/ovarian cancer families. AB - In this study we investigated 45 German breast/ovarian cancer families for germline mutations in the BRCA1 gene. We identified four germline mutations in three breast cancer families and in one breast-ovarian cancer family. among these were one frameshift mutation, one nonsense mutation, one novel splice site mutation, and one missense mutation. The missense mutation was also found in 2.8% of the general population, suggesting that it is not disease associated. The average age of disease onset in those families harbouring causative mutations was between 32.3 and 37.4 years, whereas the family harbouring the missense mutation had an average age of onset of 51.2 years. These findings show that BRCA1 is implicated in a small fraction of breast/ovarian cancer families suggesting the involvement of another susceptibility gene(s). PMID- 9391880 TI - Atypical hereditary neuropathy with liability to pressure palsies (HNPP): the value of direct DNA diagnosis. AB - We report two patients with suspected hereditary liability to pressure palsies. Neurophysiological studies showed a mixed axonal-demyelinating sensory-motor polyneuropathy with focal slowing of conduction velocities at the common sites of entrapment. Morphological studies on sural nerve biopsy from the proband showed active axonal regeneration without typical tomacula. Molecular analysis confirmed the presence of a deletion of chromosome 17p11.2 in both patients. Our observation confirms the heterogeneity of hereditary liability to pressure palsies and the relevance of DNA testing for the diagnosis of this hereditary neuropathy. PMID- 9391881 TI - Mutation and haplotype analysis of phenylalanine hydroxylase alleles in classical PKU patients from the Czech Republic: identification of four novel mutations. AB - Mutations, haplotypes, and other polymorphic markers in the phenylalanine hydroxylase (PAH) gene were analysed in 133 unrelated Czech families with classical phenylketonuria (PKU). Almost 95% of all mutant alleles were identified, using a combination of PCR and restriction analysis, denaturing gradient gel electrophoresis (DGGE), and sequencing. A total of 30 different mutations, 16 various RFLP/VNTR haplotypes, and four polymorphisms were detected on 266 independent mutant chromosomes. The most common molecular defect observed in the Czech population was R408W (54.9%). Each of the other 29 mutations was present in no more than 5% of alleles and 13 mutations were found in only one PKU allele each (0.4%). Four novel mutations G239A, R270fsdel5bp, A342P, and IVS11nt 8g-->a were identified. In 14 (5.1%) alleles, linked to four different RFLP/VNTR haplotypes, the sequence alterations still remain unknown. Our results confirm that PKU is a heterogeneous disorder at the molecular level. Since there is evidence for the gene flow coming from northern, western, and southern parts of Europe into our Slavic population, it is clear that human migration has been the most important factor in the spread of PKU alleles in Europe. PMID- 9391882 TI - Different proximal and distal rearrangements of chromosome 7q associated with holoprosencephaly. AB - Four new cases of holoprosencephaly are described in fetuses exhibiting abnormal karyotypes with different distal and proximal rearrangements of the long arm of chromosome 7. Three of them showed terminal deletions of chromosome 7q, confirming the importance of the 7q36 region in holoprosencephaly. The karyotype of the fourth fetus showed an apparently balanced de novo translocation, t(7;13) (q21.2;q33), without any visible loss of the distal part of chromosome 7q. The involvement of new genes, different from the human Sonic Hedgehog gene (hShh) responsible for holoprosencephaly, or a positional effect are discussed. PMID- 9391883 TI - Familial streptomycin ototoxicity in a South African family: a mitochondrial disorder. AB - The vestibular and ototoxic effects of the aminoglycoside antibiotics (streptomycin, gentamycin, kanamycin, tobramycin, neomycin) are well known; streptomycin, in particular, has been found to cause irreversible, profound, high frequency sensorineural deafness in hypersensitive persons. Aminoglycoside ototoxicity occurs both sporadically and within families and has been associated with a mitochondrial DNA (mtDNA) 1555A to G point mutation in the 12S ribosomal RNA gene. We report on the molecular analysis of a South African family with streptomycin induced sensorineural deafness in which we have found transmission of this same predisposing mutation. It is now possible to identify people who are at risk of hearing loss if treated with aminoglycosides in the future and to counsel them accordingly. In view of the fact that aminoglycoside antibiotics remain in widespread use for the treatment of infections, in particular for tuberculosis, which is currently of epidemic proportions in South Africa, this finding has important implications for the family concerned. In addition, other South African families may potentially be at risk if they carry the same mutation. PMID- 9391884 TI - DNA testing for fragile X syndrome: implications for parents and family. AB - The fragile X syndrome is an X linked, semidominant mental retardation disorder caused by the amplification of a CGG repeat in the 5' UTR of the FMR1 gene. Nineteen fragile X families in which the mutated FMR1 gene segregated were evaluated. The implications of the diagnosis for the parents and family were studied through pedigree information, interviews, and questionnaires. Information about the heredity of fragile X syndrome was only disseminated by family members to a third (124/366) of the relatives with an a priori risk of being a carrier of the fragile X syndrome. Twenty-six percent (94/366) of the relatives were tested. Transmission of information among first degree relatives seemed satisfactory but dropped off sharply with increasing distance of the genetic relationship, leaving 66% uninformed. This is particularly disadvantageous in an X linked disease. Of those subjects tested, 42% (39/94) had a premutation and 18% (17/94) had a full mutation. On average, in each family one new fragile X patient and two new carriers were found. When people have the task of transmitting genetic information to their relatives, they usually feel responsible and capable; however, reduced acquaintance and contact with more distant relative severely reduces the effectiveness of such transfer of information in fragile X families. PMID- 9391885 TI - Rapid identification of multiple supernumerary ring chromosomes with a new FISH technique. AB - Multiple supernumerary ring chromosomes are a rare cytogenetic finding which is poorly understood. With the introduction of FISH techniques, their chromosomal origin can now be defined clearly. The techniques described previously are complicated and time consuming. We report a new rapid technique which has been used to investigate two new cases. Multiple probes were hybridised to a single slide by means of marking the underside with a diamond pen to form a grid of squares, pipetting fixed cell suspension into the centre of each square, forming a rubber solution grid on the denatured, dehydrated slide following the lines on the underside, adding a mixture of probes into each square, and sealing the slide with a silicone rubber rim and a covering slide. The type of probe and the size, dimensions, and number of squares in the grid can be tailored to individual cases. The two new cases examined here are mosaic for three (case 1) and four (case 2) supernumerary ring chromosomes derived from different chromosomes. Normal cell lines were also present. The karyotypes were established as 47,XY,+r(4)/47,XY,+r(17)/.../48,XY,+r(17),+r(20)/ 49,XY,+r(4),+r(17),+r(20)/46,XY for case 1 and 47,XX,+r(4)/47,XX,+r(8)/47,XX,+r (10)/48,XX,+r(X),+r(4)/... /49,XX,+r(X),+r (8),+r(10)/46,XX for case 2. Our findings suggest that the ring chromosomes were formed during meiosis, perhaps involving complex rearrangements, resulting in a germ cell containing all markers, with subsequent loss of markers during cell division. Our second case also shows that the outcome is not invariably mental or physical handicap. PMID- 9391887 TI - Prenatal diagnosis of the fragile X syndrome: loss of mutation owing to a double recombinant or gene conversion event at the FMR1 locus. AB - The fragile X syndrome, an X linked mental retardation syndrome, is caused by an expanded CGG repeat in the first exon of the FMR1 gene. In patients with an expanded repeat the FMR1 promoter is methylated and, consequently, the gene is silenced and no FMR1 protein (FMRP) is produced, thus leading to the clinical phenotype. Here we describe a prenatal diagnosis performed in a female from a fragile X family carrying a large premutation. In chorionic villus DNA of the male fetus the normal maternal CGG allele and a normal pattern on Southern blot analysis were found in combination with the FRAXAC2 and DXS297 allele of the maternal at risk haplotype. A second chorionic villus sampling was performed giving identical results on DNA analysis and, in addition, expression of FMRP was shown by immunohistochemistry. We concluded that the male fetus was not affected with the fragile X syndrome. Subsequent detailed haplotype analysis showed a complex recombination pattern resembling either gene conversion or a double crossover within a 20 kb genomic region. PMID- 9391886 TI - Prader-Willi syndrome. AB - Prader-Willi syndrome is a complex disorder affecting multiple systems with many manifestations relating to hypothalamic insufficiency. Major findings include infantile hypotonia, developmental delay and mental retardation, behaviour disorder, characteristic facial appearance, obesity, hypogonadism, and short stature. Obesity and the behavioural problems are the major causes of morbidity and mortality. Prader-Willi syndrome is caused by abnormalities of the imprinted region of proximal 15q and results from absence of the normally active paternal genes in this region. Such absence results from paternal interstitial deletion, maternal uniparental disomy, or a mutation or other abnormality in the imprinting process. Diagnostic identification of all causes has become available in recent years, permitting early detection and institution of appropriate management. This testing has permitted recent identification of some phenotypic differences among affected subjects of different race and between those with deletions and uniparental disomy as a cause. PMID- 9391888 TI - Conotruncal heart defect/microphthalmia syndrome: delineation of an autosomal recessive syndrome. AB - We report on three sibs born to healthy parents, one livebirth and two terminated pregnancies, presenting with a malformation complex characterised by conotruncal heart defect (CTHD), microphthalmia, genital anomalies, and facial dysmorphism. The recurrence of the association of CTHD, particularly truncus arteriosus, and microphthalmia in sibs has previously been reported in rare instances, but a correlation between the single descriptions has never been noted. CTHDs are included among the cardiac malformations characteristically associated with the group of syndromes caused by the microdeletion of chromosome 22q11, but no detectable hemizygosity has been found in our family. An autosomal recessive gene seems to be involved in syndromic patients with the combination of CTHD and microphthalmia. The map location of this gene is at present unknown, but autosomal recessive inheritance must be considered in genetic counselling of families with children presenting with this malformation complex. PMID- 9391889 TI - Paternal transmission of congenital myotonic dystrophy. AB - We report a rare case of paternally transmitted congenital myotonic dystrophy (DM). The proband is a 23 year old, mentally retarded male who suffers severe muscular weakness. He presented with respiratory and feeding difficulties at birth. His two sibs suffer from childhood onset DM. Their late father had the adult type of DM, with onset around 30 years. Only six other cases of paternal transmission of congenital DM have been reported recently. We review the sex related effects on transmission of congenital DM. Decreased fertility of males with adult onset DM and contraction of the repeat upon male transmission contribute to the almost absent occurrence of paternal transmission of congenital DM. Also the fathers of the reported congenitally affected children showed, on average, shorter CTG repeat lengths and hence less severe clinical symptoms than the mothers of children with congenital DM. We conclude that paternal transmission of congenital DM is rare and preferentially occurs with onset of DM past 30 years in the father. PMID- 9391890 TI - Misleading linkage results in an NF2 presymptomatic test owing to mosaicism. AB - A two generation family with neurofibromatosis type 2 (NF2) is presented in which a family member requested presymptomatic molecular diagnosis. Since the consultand's mother had clinically well defined NF2, he was quoted to be at 50% risk of carrying an NF2 mutation. Mutation screening in the mother did not show the causative mutation and, consequently, presymptomatic testing was based on linkage analysis. This showed that the consultand carried the high risk chromosome 22. Subsequent mutation screening of his clinically affected sister showed a nonsense mutation, R262X in exon 8 of the NF2 gene. The mother turned out to be a mosaic for R262X; the son had not inherited the mutation. Mosaicism may be a common mechanism in NF2 and other autosomal dominant diseases with a high new mutation rate. This may be one explanation for a difference in expression in generations. Caution has to be exercised when giving results based on linkage tests which imply a very high risk to people in the second generation. PMID- 9391891 TI - Associated malformations in the family of a patient with Meckel syndrome: heterozygous expression? AB - Meckel syndrome is an inherited autosomal recessive disease. A family is described in which four persons had minor malformations related to the syndrome, suggesting the possibility of manifesting heterozygotes. It is uncertain whether these malformations represent partial expression of the disease or are coincidental. However, partial expression has been described in heterozygotes for other autosomal recessive diseases. Until the gene responsible for this lethal syndrome is cloned and sequenced, such relatives of the proband may be offered genetic counselling and prenatal diagnosis. PMID- 9391892 TI - Mucopolysaccharidosis type I: identification of novel mutations that cause Hurler/Scheie syndrome in Chinese families. AB - The complementary and genomic DNA segments of the alpha-L-iduronidase gene from two Chinese mucopolysaccharidosis type I Hurler/Scheie (MPS IH/S) patients were amplified by polymerase chain reaction (PCR) and DNA sequencing was done to study their molecular lesions. Patient W3 has heterozygous mutations; the maternal allele has M1I (G to A transition in the initiation codon ATG) and the paternal allele has Y343X (C to G transversion in exon 8 leading to in frame deletion of codons 325-343 from the mRNA owing to false splicing). Patient W2 is homozygous for mutation T364M (C to T transition in codon 364). The mutation was paternally inherited. A de novo deletion or gene conversion event may have resulted in apparent homozygosity for T364M. Expression of Y343X and T364M showed trace amounts of alpha-L-iduronidase activity compared to that of normal cDNA upon transfection into COS-7 cells. PMID- 9391893 TI - Radial aplasia and chromosome 22q11 deletion. AB - We report on a neonate with deletion 22q11 (del22q11) presenting with facial dysmorphism, ocular coloboma, congenital heart defect, urogenital malformations, and unilateral radial aplasia. This malformation complex includes features frequently occurring in velocardiofacial syndrome as well as findings described in the CHARGE and VACTERL associations. To our knowledge, the present case is the first report of radial aplasia in del22q11. This observation further supports and extends the clinical variability of del22q11. PMID- 9391894 TI - Double partial trisomy 9q34.1-->qter and 21pter-->q22.11: FISH and clinical findings. AB - We describe a patient with double trisomy 9q34.1-->qter and 21pter-->q22.1 resulting from 3:1 segregation of a maternal balanced translocation. The patient shows a clinical syndrome similar to that observed in patients with duplication of the chromosome 9q distal region, while no signs of trisomy 21 were observed. The use of high resolution banding and FISH were of fundamental importance for the cytogenetic diagnosis and for definition of the breakpoints on both chromosomes 9 and 21. PMID- 9391895 TI - Germline duplication of chromosome 2p and neuroblastoma. AB - A child with a germline duplication of chromosome 2p, 46,XY,der(13)t(2;13)(p23;q34), who developed a fatal neuroblastoma confirmed at necropsy is reported. Fluorescent in situ hybridisation studies showed chromosome 2p (p23-pter) duplicated on chromosome 13 (q34). The clinical features of the present case shared many similarities to previous reports of trisomy 2p and there have been two cases described with neuroblastoma. Germline duplication of chromosome 2p including the N-myc proto-oncogene may have pre-disposed to the development of neuroblastoma in this case. PMID- 9391896 TI - Median cleft of upper lip and pedunculated skin masses associated with de novo reciprocal translocation 46,X,t(X;16)(q28;q11.2). AB - We describe a de novo apparently balanced reciprocal translocation, 46,X,t(X;16)(q28;q11.2), in a 13 year old girl with median cleft of the upper lip, pedunculated skin masses on the nasal septum, short stature, and mental retardation. Pai syndrome is characterised by median cleft of the upper lip, pedunculated skin mass(es) on the face, and midline lipoma(s) of the central nervous system. The cause of this syndrome is unknown, although autosomal dominant inheritance has been proposed. The translocation breakpoints in the present patient may be candidate regions for a gene responsible for median cleft of the upper lip and pedunculated skin mass(es) on the face, including Pai syndrome. PMID- 9391897 TI - Rapid detection of the major deletion in the Batten disease gene CLN3 by allele specific PCR. AB - The recent isolation of the CLN3 gene involved in Batten disease (juvenile neuronal ceroid lipofuscinosis) creates possibilities for direct detection of mutations which can confirm or indicate the clinical diagnosis of Batten disease. We have designed a rapid and reliable allele specific PCR test for the detection of the major deletion, which can be used in carrier diagnosis, presymptomatic diagnosis, and prenatal diagnosis. PMID- 9391898 TI - Methionine synthase and neural tube defects. PMID- 9391899 TI - Limb-girdle muscular dystrophy or spinal muscular atrophy: a source of diagnostic confusion? PMID- 9391900 TI - The early historical roots of pediatric and adolescent gynecology. AB - BACKGROUND: The science of medicine is a constantly evolving process that builds on the experiences and observations of the past. We hypothesized that the issues of pediatric and adolescent gynecology were also of concern to physicians practicing in the 19th century. We sought to determine the extent to which our forebears of over 100 years ago considered, diagnosed, and treated these problems. METHODS: We conducted an exhaustive search through two English-language medical journals, The Obstetrical Journal of Great Britain and Ireland (vol. 1-7, 1873-1880) and The American Journal of Obstetrics (vol. 1-32, 1869-1895), for articles relating to pediatric and adolescent gynecology. RESULTS: The most frequently encountered subject was the surgical management of congenital absence or atresia of the vagina and associated anomalies of adjacent organs. By 1881, the opinion expressed by many investigators was that the method used by Thomas Addis Emmet to create an artificial vagina between the bladder and the rectum by a single-stage procedure of blunt dissection and the immediate placement of a glass vaginal dilator gave the best chance of a favorable outcome. The practice of making an artificial opening through the rectum was abandoned. In 1882, a review of published reports noted 43% postoperative mortality in children treated for benign and malignant ovarian tumors. A review article in 1891 reported a 10% mortality rate associated with treatment of the imperforate hymen. A research article in 1870 noted the mean age at menarche in England to be 14.96 years. Additional subjects in the literature included: "Acquired Venereal Disease in Children" warning of the need for "rigid ... scrutiny of the attendants and playmates of children" (1893), the enigmatic occurrence of "Vaginal Hemorrhage in an Infant Five-Days-Old" (1874), the brutal atrocities perpetrated against "The Child Wives of India" (1895), "Early Pregnancy" reviewing childbirth by young adolescents (1874), "Hermaphrodism" (1886), "The ... Hymen and Its Remains ..." (1871), a "Case of Fatal Hemorrhage from the Genital Organs" in which a 17-year old exsanguinated from a vaginal laceration (1879), and "Primary Sarcoma of the Vagina ... in a Child Three-Years-Old" (1881). CONCLUSION: Modern pediatric and adolescent gynecology can trace its roots to well over a century ago. PMID- 9391901 TI - Use of hormonal methods of birth control among sexually active adolescent girls. AB - OBJECTIVE: To identify factors associated with the use of various birth control methods among sexually active adolescent girls. DESIGN: A survey distributed as part of a larger study measuring compliance with hepatitis B vaccination. SETTING: A hospital-based and a school-based clinic. MEASURES: Demographic and health behavior data including sexual activity, contraceptive method, substance use, condom use, and history of sexually transmitted diseases (STDs) were collected. Birth control method was confirmed by medical record review. Associations with the outcome variable of birth control method were analyzed using chi square, Kruskal-Wallis analyses of variance, and t-tests, followed by logistic regression analysis. RESULTS: Among sexually experienced girls, 39% (n = 123) reported using oral contraceptive pills (OCPs), 5.4% (n = 17) used Depo Provera (medroxyprogesterone acetate) or Norplant (levonorgestrel), and 55.6% (n = 175) used no hormonal method. Logistic regression analysis revealed that the factors most significantly associated with the use of hormonal methods were older age (odds ratio [OR] = 1.19; 95% confidence interval [CI], 1.07-1.33), not using a condom at last intercourse (OR = 0.55; CI, 0.34-0.90), and having had a well visit within 1 year (OR = 2.11; CI, 1.12-3.70). OCP users were less likely than Depo-Provera or Norplant users to have used alcohol (p = 0.041), cigarettes (p = 0.002), or marijuana (p = 0.018) in the past 30 days. OCP users were less likely than nonusers of hormonal methods to have smoked cigarettes (p = 0.034) or marijuana (p = 0.052). The school-based clinic had a greater proportion of subjects using long-acting progestins (p < 0.001). CONCLUSIONS: The decreased rate of condom use among those who used hormonal birth control methods and the different rates of health risk behaviors among users of various methods require targeted counseling efforts to decrease pregnancy and STD rates among young women. PMID- 9391902 TI - Prevalence of endometriosis in adolescent girls with chronic pelvic pain not responding to conventional therapy. AB - STUDY OBJECTIVE: To evaluate adolescent girls with chronic pelvic pain not responding to conventional medical therapy, using advances in operative laparoscopy to determine endometriosis prevalence, clinical stage, and type of lesion. DESIGN: A descriptive retrospective study of subjects who (1) were referred for the evaluation of chronic pelvic pain, (2) did not respond to a nonsteroidal anti-inflammatory drug and an oral contraceptive pill, and (3) underwent a laparoscopy to determine the etiology of the pelvic pain. SETTING: Patients referred to a surgical gynecologist in a pediatric/adolescent gynecology and reproductive endocrine academic practice. PARTICIPANTS: All patients younger than 22 years of age with chronic pelvic pain. INTERVENTION: Operative laparoscopy to determine the etiology of the chronic pelvic pain. MAIN OUTCOME MEASURES: Operative laparoscopy results including stage and description of endometriosis. RESULTS: More than two thirds of the study population (69.6%) was found to have endometriosis. All subjects had either stage I or II as determined by the American Fertility Society's classification system. The nature of the pain in the 32 subjects with endometriosis was both acyclic and cyclic in 20 (62.5%), acyclic only in 9 (28.1%), and cyclic only in 3 (9.4%). Other presenting symptoms included gastrointestinal in 11 (34.3%), urinary in 4 (12.5%), and irregular menses in 3 (9.4%). CONCLUSIONS: Adolescents with chronic pelvic pain not responding to medical therapy have a high rate of endometriosis and should be referred to a gynecologist who is experienced with the subtle laparoscopic findings of atypical endometriosis to diagnose the etiology of the pelvic pain and initiate appropriate therapy. PMID- 9391903 TI - Attitudes of female college students toward over-the-counter availability of oral contraceptives. AB - STUDY OBJECTIVE: To determine female college students' beliefs about oral contraceptive pill (OCP) availability and use, and to examine significant factors associated with these beliefs. DESIGN: Cross-sectional survey. SETTING: Urban women's liberal arts college. PARTICIPANTS: Two hundred ninety female undergraduates who completed surveys. INTERVENTION: An anonymous survey was placed in all undergraduate mailboxes. Surveys were returned to a locked collection box in the mailroom. Within 4 weeks after distribution. 290 surveys were completed. MAIN OUTCOME MEASURES: Sexual and contraceptive practices and students' beliefs regarding whether OCPs should be made available without prescription. RESULTS: The respondents' average age was 20.9 +/- 3.3 years; 84% reported previous sexual intercourse with the mean age of first intercourse at 16.6 +/- 2.2 years. Seventy-five percent of the sexually active women reported use of OCPs and 52% had used OCPs at their last intercourse. Sixty-five percent of all respondents felt OCPs should not be available without prescription. The two most commonly cited reasons for not wanting OCPs to be available over the counter (OTC) were that (1) side effects might occur that a health care provider could have prevented (59%), and (2) people would not go to their providers for regular check ups (56%). The most commonly cited reason for believing that OCPs should be available OTC was that there would be fewer unwanted pregnancies (74%). Race, previous OCP use, previous sexual activity, and perceived risk of pregnancy were not significant predictors of believing OCPs should be available OTC. Having had a previous pregnancy was a significant predictor of believing OCPs should be available OTC (p = 0.047). Those who believed OCPs should be available only with a prescription were willing to pay more for OCPs (p = 0.033). Logistic regression controlling for race revealed that both younger age (p = 0.03) and previous pregnancy (p = 0.002) were independent predictors of believing OCPs should be available OTC. CONCLUSIONS: The majority of our sample believe that OCPs should remain as a prescription medication. Previous pregnancy and younger age are important factors in determining beliefs regarding OCP availability. Further studies in a more diverse population are needed to explore the relationship between age, previous pregnancy, and desire for contraceptive availability without prescription. PMID- 9391904 TI - Adolescent girls' understanding of Papanicolaou smear results. AB - STUDY OBJECTIVE: Human papillomavirus (HPV) urogenital infections are common in sexually active adolescents. Previous research has indicated that adolescent girls do not reliably report histories of HPV infection. This study examined whether asking an adolescent girl if she had ever had an abnormal Papanicolaou (PAP) smear was a good screening question for evidence of HPV urogenital infection. DESIGN: The responses to the question about abnormal PAP smears, were compared with their charts for documented abnormal PAP smear, HPV infection, and sexually transmitted infection. SETTING: An urban, hospital-based adolescent clinic. PARTICIPANTS: Fifty adolescent girls (mean age, 14.8 years). MAIN OUTCOME MEASURES: Degree of agreement (kappa statistic). RESULTS: Using a kappa statistic, reported history of an abnormal PAP smear had "fair" agreement with documented dysplasia on PAP smear and "moderate" agreement with documented HPV infection (i.e., either condyloma on PAP smear or genital warts noted on examination). This reported history of an abnormal PAP smear agreed better with documented HPV infection than with documented dysplasia on PAP smear. CONCLUSIONS: There appears to be considerable confusion among adolescent girls regarding their PAP smear results. Care providers need to be sensitive to this when they are collecting historical information and when they are diagnosing HPV infection or an abnormal PAP smear. PMID- 9391905 TI - Congenital cervicovaginal aplasia with septate uterus and functioning endometrium. AB - A 14-year-old adolescent girl presented with primary amenorrhea and uncontrolled pelvic pain. Evaluation using pelvic sonogram, magnetic resonance imaging, and laparoscopy confirmed the diagnosis of cervicovaginal aplasia with functioning endometrium and a vaginal fistulous tract. At age 19 years, hysterectomy and vaginoplasty allowed the patient to be free of pain and to have normal sexual function. PMID- 9391906 TI - Non-Hodgkin's ovarian lymphoma during adolescence: report of two cases. AB - STUDY OBJECTIVE: To present two cases of non-Hodgkin's ovarian lymphoma during adolescence. DESIGN: Follow up of the patients. Report of diagnostic and therapeutic approaches. SETTING: Division of Pediatric and Adolescent Gynecology and Corrective Gynecological Surgery, University of Athens (Athens, Greece). INTERVENTIONS: Laparotomy and chemotherapy. RESULTS AND MAIN OUTCOME: Three- and five-year survival rates. CONCLUSIONS: Ovarian lymphoma constitutes a rare entity with guarded prognosis. Selective surgery and chemotherapy constitute the treatment of choice. PMID- 9391907 TI - Design, synthesis and conformational analysis of hGM-CSF(13-31)-Gly-Pro-Gly-(103 116). AB - On the basis of the X-ray structure and results from structure-activity relationship studies, the following GM-CSF analogue was designed and synthesized by solid-phase methodology: hGM-CSF[13-31]-Gly-Pro-Gly-[103-116]-NH2. This analogue was constructed to comprise helices A and D of the native hGM-CSF, covalently linked in an antiparallel orientation by the tripeptide spacer Gly-Pro Gly, which is known as a turn-inducing sequence. The conformational analysis of the analogue by CD spectroscopy revealed an essentially random structure in water, while alpha-helix formation was observed upon addition of TFE. In 40% TFE the helix content was approximately 45%. By two-dimensional NMR experiments in 1:1 water/trifluoroethanol mixture two helical sequences were identified comprising the segments corresponding to helix A and helix D. In addition to medium-range NOESY connectivities, a long-range cross-peak was found involving the leucine residues at positions 13 and 35. Based on the experimentally derived data (54 NOEs), the structure was refined by restrained molecular dynamics simulations over 120 ps at various temperatures. A representative conformation derived from the computer simulation is mainly characterized by two helical segments connected by a loop region. The overall three-dimensional structure of the analogue is comparable to the X-ray structure of hGM-CSF in that helices A and D are oriented in an antiparallel fashion, forming a two alpha-helix bundle. Nevertheless, there are small differences in the topology of the helices between the solution structure of the designed analogue and the X-ray structure of hGM CSF. The possible implications of these conformational features at the effects of biological activity are discussed. PMID- 9391908 TI - Conformation of four peptides corresponding to the alpha-helical segments of human GM-CSF. AB - The conformation of segments corresponding to the four alpha-helical stretches found in human granulocyte-macrophage colony-stimulating factor was studied in water solution in the presence of different amounts of 2,2,2-trifluoroethanol (TFE). The CD spectra reveal the onset of secondary structure upon addition of TFE. The final amount of helical conformation varies among the four peptides. In all cases, the conformational transition is complete before 50% TFE (v/v). 1H-NMR studies were conducted at this solvent composition, leading to the assignment of all the resonances and to the definition of the secondary structure for all four fragments. PMID- 9391910 TI - Optically active aromatic amino acids. Part V: Some N-t-butyloxycarbonyl-O-methyl L-tyrosine analogues with ring substitution at position 3. PMID- 9391909 TI - Synthesis and CD studies of an 88-residue peptide containing the main receptor binding site of HTLV-I SU-glycoprotein. AB - Essential HTLV-1 biological functions, like host-cell receptor recognition, depend on the structural motives on the surface glycoprotein gp46. We defined a peptide of 88 amino acids [Arg147-Leu234] corresponding to the central part of the protein sequence, where major neutralizing epitopes are localized. After evaluating the feasibility of its chemical synthesis, the chosen sequence was realized using the stepwise solid-phase methodology. Multiple chromatographic purification steps were required to obtain a sample suitable for structural analysis. Correct folding was supported by strong binding of monooclonal antibodies, recognizing known exposed immunodominant regions. Circular dichroism studies confirmed a non-random conformation of at least 70-80% of the synthetic peptide. Investigation of the 3D-structure of the synthetic peptide will provide useful information for future vaccine and drug-design strategies. PMID- 9391911 TI - Enzymatic synthesis of isotopically labelled serine and tryptophan for application in peptide synthesis. AB - L-[1.2-13C2, 15N]Serine was prepared from [1,2-13C2, 15N]glycine on a gram scale by the use of the enzyme serine hydroxymethyltransferase. The reaction was monitored by 13C-NMR spectroscopy. This is the first simultaneously 13C- and 15N labelled serine isotopomer so far reported. Part of the product was directly converted by tryptophan synthase to L-[1,2-13C2, 15N]tryptophan which could conveniently be purified and isolated as Boc-derivative in a yield of 71%. Most of the serine was isolated similarly but to remove remaining starting material in this case purification by column chromatography was required. PMID- 9391912 TI - Conformational characterization of peptides rich in the cycloaliphatic C alpha,alpha-disubstituted glycine 1-aminocyclononane-1-carboxylic acid. AB - A series of N- and C-protected, monodispersed homo-oligopeptides (to the pentamer level) from the cycloaliphatic C alpha,alpha-dialkylated glycine 1 aminocyclononane-1-carboxylic acid (Ac9c) and two Ala/Ac9c tripeptides have been synthesized by solution methods and fully characterized. The conformational preferences of all the model peptides were determined in deuterochloroform solution by FT-IR absorption and 1H-NMR. The molecular structures of the amino acid derivatives mCIAc-Ac9c-OH and Z-Ac9c-OtBu, the dipeptide pBrBz-(Ac9c)2-OtBu, the tetrapeptide Z-(Ac9c)4-OtBu, and the pentapeptide Z-(Ac9c)5-OtBu were determined in the crystal state by X-ray diffraction. Based on this information, the average geometry and the preferred conformation for the cyclononyl moiety of the Ac9c residue have been assessed. The backbone conformational data are strongly in favour of the conclusion that the Ac9c residue is a strong beta-turn and helix former. A comparison with the structural propensity of alpha aminoisobutyric acid, the prototype of C alpha,alpha-dialkylated glycines, and the other extensively investigated members of the family of 1-aminocycloalkane-1 carboxylic acids (Acnc, with n = 3-8) is made and the implications for the use of the Ac9c residue in conformationally constrained analogues of bioactive peptides are briefly examined. PMID- 9391914 TI - The regiospecific N-derivatization of histidine side chains: reinvestigation of a supposed N tau to N pi migration. AB - A report claiming that AcHisOMe reacts regiospecifically with 4 fluoronitrobenzene to give AcHis[tau Ph(NO2)]OMe, which on treatment with H2/Pd(C) undergoes a partial tau-pi shift to give some AcHis[pi Ph(NH2)]OMe, cannot be substantiated. 4(5)-Methylimidazole, a model for AcHisOMe, gives on reaction with 4-fluoronitrobenzene a 4:1 mixture of the regioisomers 1-(4 nitrophenyl),4-methylimidazole, corresponding to tau-substitution, and 1-(4 nitrophenyl),5-methylimidazole, corresponding to pi-substitution, each of which has been isolated and fully characterized, including proof of orientation. In both cases, treatment with H2/Pd(C) gives a single product, without any change of orientation in either case. PMID- 9391913 TI - The design of a specific ligand of HIV gp120. AB - The crystal structure of CD4 suggested that the C/G38 and C/L44 replacements with the consequent cystine bridge formation are compatible with the native structure of that molecular moiety. As the NQGSF sequence, corresponding to the 39-43 fragment of human CD4 protein, was found to be involved in the HIV gp120 interaction, it has been synthesized in a cyclic form by adding two cysteine residues at the amino and carboxy termini. 1H-NMR studies show that the predominant solution conformation of cyclo-[CNQGSFC] is a type II beta-turn centred on the NQGS segment. Structural and dynamic properties of the peptide are also analysed in relation to the in vitro activity. PMID- 9391915 TI - Prevalence, incidence and stability of drinking problems among whites, blacks and Hispanics: 1984-1992. AB - OBJECTIVE: This article reports on the prevalence, incidence and stability of dependence-related problems and social consequences from drinking among whites, blacks and Hispanics between 1984 and 1992. METHOD: A probability sample of 1,777 whites, 1,947 blacks and 1,453 Hispanics from the U.S. adult household population was interviewed in 1984. In 1992 a subsample consisting of 788 whites, 723 blacks and 703 Hispanics was reinterviewed. Interviews averaging 1 hour in length were conducted in respondents' homes by trained interviewers. RESULTS: The prevalence of a number of alcohol-related problems, the stability and incidence of dependence-related problems and the incidence of social consequences from drinking are higher among Hispanic than among white men. Dependence-related problems are more stable among black than among white men. Among women, the incidence of dependence-related problems and social consequences from drinking is higher among blacks than whites. Hispanic women have a higher incidence of social consequences from drinking than white women. Having a problem at Time 1 correlates only moderately with having a problem at Time 2, independent of ethnicity. CONCLUSIONS: In general, Hispanics and blacks continue to be more at risk than whites for developing a number of alcohol-related problems. These two ethnic groups should be the focus of renewed efforts to address alcohol problems and inequalities in income distribution, employment, education and lack of access to adequate health care. PMID- 9391916 TI - Mate similarity, heavy substance use and family history of problem drinking among young adult women. AB - OBJECTIVE: This study used data from a national sample of young adult women to evaluate issues about spousal similarity for problem drinking. Paternal and maternal problem drinking were also evaluated in regard to daughters' marriage to a problem drinking spouse, and daughters' problem drinking and substance use. METHOD: Data from over 5,000 young adult women (ages 23-30 yrs) from the National Longitudinal Survey of Youth (NLSY) archive were used to evaluate associations between marrying a problem-drinking spouse, family history of problem drinking, and women's problem drinking and lifetime marijuana and cocaine use. RESULTS: Findings indicated that black women were less likely to marry a problem-drinking spouse than were Hispanic, Native American or white women. Problem-drinking women were twice as likely to have married a problem-drinking spouse than were non drinking women, and heavier lifetime marijuana or cocaine use by women was also associated with an almost twofold increase in marrying a problem-drinking spouse. Random effects ordinal probit regression models indicated that, while controlling for major sociodemographic variables (e.g., race, poverty status), maternal, paternal and spousal problem drinking all significantly predicted problem drinking and heavier levels of substance use among the women. CONCLUSIONS: Nonrandom matching of problem drinking among marital partners was indicated in this study and women's problem drinking and substance use practices were predicted by paternal, maternal and spousal problem drinking. The similarity of problem-drinking spouses was not constant across racial/ethnic groups, as black women were less likely to marry a problem-drinking spouse, though racial differences in the age of onset of heavier drinking may have influenced this finding. PMID- 9391917 TI - Periods of abstinence following the onset of alcohol dependence in 1,853 men and women. AB - OBJECTIVE: Data from both laboratory experiments and retrospective histories of alcoholics indicate that alternations between periods of abstinence and heavy drinking are common in the course of alcoholism. This article examines the prevalence, chronological characteristics and clinical features associated with periods of abstinence in a large sample of alcohol dependent men and women. METHOD: As part of the Collaborative Study on the Genetics of Alcoholism (COGA), semistructured personal interviews were used to gather data on the course of alcoholism in 1,853 alcohol dependent men and women. Data were evaluated regarding the characteristics of alcoholics with and without periods of abstinence lasting 3 or more months, and the course of these periods over time were evaluated separately for subjects with one, two, three or four episodes of abstention. RESULTS: Despite a relatively high threshold of 3 months for defining an abstinence period, 62.3% of the subjects had at least one such episode, including 55.6% of the 1,853 alcoholics for whom complete data were available. Those alcohol dependent subjects with periods of abstention had an average (+/- SD) of 1.7 +/- 0.99 such periods, and 43% of abstainers had two or more. Logistic regression analyses revealed that the predictors of history of abstention were female gender (odds ratio [OR] = 1.78), older age at interview (OR = 1.04 per year), younger age of onset of alcoholism (OR 0.93 per year), ever having been married (OR = 1.45), the number of DSM-III-R dependence items endorsed (OR = 1.03 per item) and attendance in AA (OR = 2.82). Among abstainers, a period lasting 5 or more years was predicted by older age (OR = 1.03 per year) and AA participation (OR = 3.23), but also by more years of alcoholism (OR = 1.06 per year), white racial background (OR = 1.79) and the absence of history of having been a proband (OR = 3.39). CONCLUSIONS: Periods of abstinence of 3 or more months were commonly observed in the course of alcohol dependence. This information is important for clinicians who need to avoid inappropriate stereotypes of alcoholism and who wish to educate their alcohol dependent patients about the course of this disorder. PMID- 9391918 TI - Sex and strain influence the effect of ethanol on central monoamines. AB - OBJECTIVE: We recently investigated the effects of EtOH on the mesolimbic dopamine and serotonin systems in male and female C57BL/6 (B6) and DBA/2J (D2) mice. METHOD: Male and female rodents from the B6 and D2 mouse strains (n = 11 per strain, sex and dose) were used in this study. Doses of EtOH (vs saline) administered were 1.0, 2.0 or 3.0 g/kg. RESULTS: Treatment with saline or EtOH produced both strain- and sex-dependent differences in patterns of monoamine response. For example, D2s exhibited significantly higher overall dopamine (DA) levels than did B6s in the frontal cortex (FC), nucleus accumbens (NA) and caudate-putamen (CP). In the FC, female D2 evinced elevated 5HIAA at 1.0 g/kg. In the NA, D2 females showed dose related increases in levels of DA up to 3.0 g/kg, whereas in the D2 males and in B6 males and females we observed no response. Also in the NA, B6 males showed increases in dihydroxyphenyacetic acid (DOPAC) at 1.0 and 3.0 g/kg. In the CP, B6 males showed higher DA levels than B6 females at the saline, and all EtOH doses. For serotoninergic activity in the CP as well as the NA, EtOH produced a distinctive triphasic response, with the 1.0 and 3.0 g/kg doses of EtOH producing higher levels than saline and 2.0 g/kg of 5HIAA in B6 males than in B6 females. CONCLUSIONS: Our findings indicate strain and sex differences in monoamine response to acute doses of ethanol, and further implicate (via changes in DOPAC) presynaptic mechanisms in the effects of ethanol on dopamine. PMID- 9391919 TI - Alcohol and the ability to inhibit behavior in men and women. AB - OBJECTIVE: This experiment tested the impairing effect of alcohol on cognitive inhibitory control of behavior in the absence of any motivational consequences for exhibiting or inhibiting a response. METHOD: Men (n = 24) and women (n = 24) were trained on a computerized "go-stop" task that measured response reaction time (RT) to a go signal and inhibitory control by the number of inhibitions to a randomly occurring stop signal. Equal numbers of men (n = 8) and women (n = 8) were assigned to one of three groups (n = 16), and they performed the task alone in a room under either alcohol (A), placebo (P) or no-treatment control (C) conditions. Blood alcohol concentrations of men and women were matched in Group A by administering 0.62 and 0.54 g/kg of alcohol, respectively. RESULTS: Alcohol impaired inhibitory control and had no significant effect on response RT. Under P and C conditions, no changes in inhibitions or response RT were observed. In addition, no significant gender effects were found. CONCLUSIONS: The results showed that inhibitory control of behavior was impaired by a moderate dose of alcohol that did not affect response RT. PMID- 9391920 TI - P300 from men with a family history of alcoholism under different incentive conditions. AB - OBJECTIVE: Males with a family history of alcoholism (Family History Positive [FHP]) have frequently been reported to show an attenuated amplitude of the P300 component of the EEG event-related potential (ERP). The purpose of the present study was to explore the influence of incentives on the amplitude of the P300 to Target stimuli in FHP and FHN (Family History Negative) men. METHOD: The ERPs of 20 FHP and 20 FHN men were recorded in a visual discrimination task under two conditions: a no incentive (Neutral) and a reward/loss of reward (Incentive) conditions. ERPs following Target, Non-Target and Novel stimuli were examined. RESULTS: The FHP subjects displayed the expected attenuation of P300 amplitude, regardless of the stimulus type (Target, Non-Target or Novel), compared to the FHN subjects. The FHN subjects showed the predicted effect of a significantly increased P300 amplitude following Target stimuli in the Incentive condition whereas the FHP subjects did not display significantly greater P300 amplitudes in response to the incentive. CONCLUSIONS: These results may reflect a deficit in the motivational-cognitive system of FHP subjects. Analyses suggest that changes in P300 amplitude in response to the incentive can be predicted by subjects' scores on self-report measures of sensation seeking and behavioral undercontrol. However, such measures could not explain subjects' absolute P300 amplitude to Target stimuli. PMID- 9391921 TI - Stability of neuropsychological assessments early in alcoholism treatment. AB - OBJECTIVE: Current trends in managed mental health care have telescoped the assessment and treatment of individuals diagnosed with an alcohol or other drug use disorder. Yet, there is limited empirical information about the short-term stability of neuropsychological status and other person characteristics that are useful to assess early in treatment. This study examined the stability of neuropsychological test scores within the first 3 weeks following diagnosis of an alcohol use disorder. METHOD: An eclectic neuropsychological battery made up of commonly used, sensitive tests of abstraction, executive functions, memory, visuospatial abilities and verbal ability was administered to female and male alcohol use disordered individuals within 3 days of treatment entry (or following detoxification), 3-5 days later and 21 days later. The three test administrations were completed by 35, 32 and 24 subjects, respectively. RESULTS: Across tests, the average stability coefficient (Pearson correlation) was .82 between Days 3 and 5, .86 between Days 5 and 21, and .79 between Days 3 and 21. Intraclass correlations ranged from .79 to .98 across tests (mean = .92). Clinical stability, defined as the likelihood that a test score fell consistently above or below a standardized impairement cutoff score, was also good. Across tests, percent agreement in impairment diagnoses for the same three time intervals averaged 84%, 92% and 87%, respectively. The chance-corrected kappa (Kappa) coefficients of diagnostic agreement were generally moderate to substantial from Day 3 to Days 5 or 21, and mostly substantial from Day 5 to 21. CONCLUSIONS: Early assessments of neuropsychological status were psychometrically stable, and also provided reasonably stable indicants of clinically significant impairment. It was likely that the data provided lower bound estimates of the stability of impairment classifications due to the repeated measures design and power limitations. PMID- 9391922 TI - The effect of question structure on self-reports of heavy drinking: closed-ended versus open-ended questions. AB - OBJECTIVE: We compared open-ended versus closed-ended questions on the frequency of consuming five or more drinks in a single sitting. METHOD: From a general population survey of Ontario adults (N = 2,022, 62% male), we analyzed a subsample of 649 respondents who reported drinking five or more drinks in a single sitting at least once in the past year. Differences in agreement between the two questions and rates of missing data were evaluated. RESULTS: For the most part, the two measures were not consistent, with the closed-ended question eliciting higher rates of heavier drinking. Rates of missing data were also higher for the open-ended question. CONCLUSIONS: Open-ended question may not necessarily be more suitable than closed-ended questions for estimating the frequency of heavy alcohol use. PMID- 9391923 TI - Relationship between inpatient alcoholism treatment and longitudinal changes in health care utilization. AB - OBJECTIVE: The purpose of the study was to evaluate changes in health care utilization associated with inpatient alcoholism treatment in alcoholics of low socioeconomic status with different histories of treatment relapse. METHOD: The sample consisted of more than 85,000 male alcoholics using inpatient care in Department of Veterans Affairs medical centers in fiscal year 1987. Five treatment groups were identified to represent a continuum of length and intensity of alcoholism treatment, including formal inpatient alcoholism treatment, short detoxification and hospitalizations for primary diagnoses other than alcoholism. All inpatient and outpatient health services for 3 years before and 3 years after the index hospitalization were examined for differential changes in utilization associated with the five treatment groups after controlling for patient predisposing, enabling and need characteristics. RESULTS: Both total inpatient days and outpatient visits increased significantly for all treatment groups, with the greatest increases occurring in the group completing inpatient alcoholism treatment (both p < .0001). However, use of inpatient medical care decreased and substance abuse inpatient care increased significantly for most groups, with the largest increases in substance abuse care found for the completed treatment group. CONCLUSIONS: In a hospital system that does not deny care on the basis of ability to pay, certain groups of chronic alcoholics who cannot sustain prolonged remission will continue to be heavy utilizers of services. Alcoholism treatment may be associated with higher short-term costs but it remains to be seen whether provision of more focused treatment services is able to achieve longer term better outcomes and, ultimately, lower costs. PMID- 9391924 TI - Age of first use: its reliability and predictive utility. AB - OBJECTIVE: The purpose of this study was: (1) to assess the utility of age of first licit use and age of first illicit use as predictors of alcohol and drug use at ages 20 and 30; and (2) to examine the reliability of retrospectively recalled ages of onset of use. METHOD: Subjects (N = 839) from the Rutgers Health and Human Development Project provided four waves of longitudinal data spanning the age range from 15 to 31. RESULTS: Retrospective recall of age of onset revealed a fair degree of relative agreement but a lack of absolute agreement because of an upward shift in recalled ages as individuals became older. Repeated measures ANOVAS revealed normative declines in alcohol and drug use from 20 to 30 even though individual differences in use remained quite stable across time. Regression analyses indicated that: (1) age of first licit use as recalled at age 18 did not predict alcohol or drug use at age 20; (2) age of first illicit use was a weak predictor of alcohol use at 20 but a fairly strong predictor of drug use at 20; and (3) neither age predicted use or use consequences at age 30. CONCLUSIONS: In the general population, illicit drug use and heavier alcohol use are, regardless of age of onset, adolescence-limited phenomena for most individuals. Findings suggest that intervention efforts need to be aimed simultaneously at delaying the onset of illicit use and reducing use levels among young adult users. PMID- 9391925 TI - Stress, alcohol-related expectancies and coping preferences: a replication with adolescents of the Cooper et al. (1992) model. AB - OBJECTIVE: The present study attempted to replicate with adolescents the stressor vulnerability model of adult drinking proposed by Cooper et al. (J. Abnorm. Psychol. 101: 139-152, 1992). The Cooper et al. model simultaneously assesses the stress-moderating effects of gender, expectancies and coping on alcohol use and abuse. METHOD: Adolescents in Grades 7-12 (N = 184, 59% female) completed the Alcohol Expectancy Questionnaire-Adolescent form, the COPE, the Adolescent Perceived Events Scale and the Drinking to Cope scale. RESULTS: The pattern of results was very similar to those of earlier studies using adults or undergraduates. Generally, positive expectancies for alcohol, an avoidant coping preference and stress were predictive of drinking to cope, alcohol use and alcohol-related problems. A number of two-way interactions were also reported. Although gender did not play a prominent role in prediction, as it typically does with adults, grade was a significant predictor; older students reported more alcohol use and alcohol-related problems than younger students. CONCLUSIONS: Results were similar to those reported by Cooper et al. with adults and Evans and Dunn (J. Stud. Alcohol 56: 186-193, 1995) with undergraduates, and support the utility of the stressor vulnerability model for understanding alcohol use among adolescents. PMID- 9391927 TI - Blood alcohol concentration and driver record factors. PMID- 9391926 TI - Risk and protective factors as predictors of adolescent alcohol involvement and transitions in alcohol use: a prospective analysis. AB - OBJECTIVE: Determinants of initial alcohol use may differ from predictors of accelerated or problematic consumption. Social influences may be strong predictors of initial drinking; however, later stages of problem drinking may be linked developmentally to intrapersonal deficits. This study prospectively examined the influence of chronic and changing risk and protective status in predicting adolescent alcohol involvement and transitions in alcohol use. METHOD: Data were obtained from a three-wave cohort (N = 823) of 8th-10th grade nonintervention students participating in a school-based drug abuse prevention trial. Cognitive, attitudinal and social influence measures were dichotomized using empirical cut-offs to designate risk or protective status. Using a conceptually based assignment scheme, additive risk indices were created assessing chronic (averaging across time) and changing features of competence, psychological and interpersonal functioning, cognitive-affective and social influences. Three chronic and change protective indices were created tapping competence, psychological, and interpersonal functioning. RESULTS: Controlling for initial drinking and gender, chronic risk for social influence and psychological functioning and increased risk for social influences and competency predicted subsequent drinking behavior. Chronic psychological protection attenuated subsequent drinking. Using categorical measures of drinking behavior to designate nonuse, experimental or moderate-heavy use, chronic social influence and competency risk were associated with an increased likelihood of accelerated drinking, whereas improved psychological functioning diminished the likelihood of increased drinking behavior. CONCLUSIONS: Findings underscore the need for implementing prevention strategies that reinforce developmentally appropriate skills and enhance personal competence and psychological functioning as effective barriers against initial and more problematic alcohol use. The unique contribution of protective forces also underscores that risk reduction and protection enhancement are complementary processes and are both required to offset social influences for alcohol consumption. PMID- 9391928 TI - Telemedicine and the national information infrastructure: are the realities of health care being ignored? AB - Health care is shifting from a focus on hospital-based acute care toward prevention, promotion of wellness, and maintenance of function in community and home-based facilities. Telemedicine can facilitate this shifted focus, but the bulk of the current projects emphasize academic medical center consultations to rural hospitals. Home-based projects encounter barriers of cost and inadequate infrastructure. The 1996 Telecommunications Act as implemented by the Federal Communications commission holds out significant promise to overcome these barriers, although it has serious limitations in its application to health care providers. Health care advocates must work actively on the federal, state, and local public and private sector levels to address these shortcomings and develop cost effective partnerships with other community-based organizations to build network links to facilitate telemedicine-generated services to the home, where the majority of health care decisions are made. PMID- 9391929 TI - The virtual visit: using telecommunications technology to take care of patients. AB - Telephone-Linked Care (TLC) technology has been developed and applied as an alternative to and a supplement for office visits as a means to deliver ambulatory care. TLC is used to monitor patients with chronic diseases, counsel patients on important health behaviors, and provide information and support to home caregivers of patients with disabling conditions. TLC speaks to patients over the telephone in their homes using computer-controlled digitized human speech. Patients use their telephone keypad to communicate. TLC conversations last 2-15 minutes per call and take place weekly for periods of at least 3 months. The conversations consist of a salutation, password verification, the core clinical part, and a closing. The structure of the clinical part is similar for each of the application groups: chronic disease, health behavior, and caregiver support. The system architecture consists of linked voice and database components and their subcomponents. Preliminary evaluation indicates that TLC is well accepted by patients and their providers and can improve clinical outcomes. PMID- 9391930 TI - Quality-of-life research on the Internet: feasibility and potential biases in patients with ulcerative colitis. AB - OBJECTIVE: The World Wide Web (WWW) is a new communications medium that permits investigators to contact patients in nonmedical settings and study the effects of disease on quality of life through self-administered questionnaires. However, little is known about the feasibility and, what is more important, the validity of this approach. An on-line survey for patients with ulcerative colitis (UC) and patients whose UC had been treated with surgical procedures was developed. To understand how patients on the WWW might differ from those in practice and the potential biases in conducting epidemiological research in volunteers recruited on the Internet, post-surgery patients who responded to the WWW survey were compared with those in a surgical practice. SETTING: The Internet and private practice surgical clinic. MAIN OUTCOMES: Scores from the Short form 36 (SF-36) Health Assessment Questionnaire and the Self-Administered Inflammatory Bowel Disease Questionnaire (IBDQ). RESULTS: Over a 5-month period, 53 post-surgery patients enrolled in the Internet study; 47 patients from a surgical clinic completed the same computer-based questionnaire. Surgically treated patients on the WWW were younger than their clinic counterparts (median age category 35-44 years vs. 45-54 years, p = 0.01) but more ill with a lower summary IBDQ score (168 vs. 186, p = 0.019) and lower health status across almost all dimensions of the SF-36 (p = 0.016). CONCLUSIONS: It is feasible to conduct epidemiological research on the effects of UC on quality of life on the Web; however, systematic differences in disease activity between volunteer patients on the WWW and "in the clinic" may limit the applicability of results. PMID- 9391931 TI - A voice-enabled, structured medical reporting system. AB - Kurzweil Applied Intelligence received a research grant from the National Institute of Standards and Technology (NIST) Advanced Technology Program to develop a prototype voice-enabled, structured medical reporting system. In typical usage, the physician dictates to the system, which then uses automatic speech recognition and medical knowledge bases to produce a structured report. This report can then be formatted and viewed on a computer screen, stored in databases of patient information, transmitted to other systems, used to support outcome studies, or viewed on a Web browser. The output reports are structured according to two standard, platform-independent formats: SGML and CORBA. These formats represent the data in a way that can be read by both computers and humans, and efficiently communicated to a wide range of databases and communications protocols. PMID- 9391933 TI - A WWW implementation of national recommendations for protecting electronic health information. AB - In March of 1997, the National Research Council (NRC) of the National Academy of Sciences issued the report, "For the Record: Protecting Electronic Health Information." Concluding that the current practices at the majority of health care facilities in the United States are insufficient, the Council delineated both technical and organizational approaches to protecting electronic health information. The Beth Israel Deaconess Medical Center recently implemented a proof-of-concept, Web-based, cross-institutional medical record, CareWeb, which incorporates the NRC security and confidentiality recommendations. We report on our WWW implementation of the NRC recommendations and an initial evaluation of the balance between ease of use and confidentiality. PMID- 9391932 TI - Recommendations for responsible monitoring and regulation of clinical software systems. American Medical Informatics Association, Computer-based Patient Record Institute, Medical Library Association, Association of Academic Health Science Libraries, American Health Information Management Association, American Nurses Association. AB - In mid-1996, the FDA called for discussions on regulation of clinical software programs as medical devices. In response, a consortium of organizations dedicated to improving health care through information technology has developed recommendations for the responsible regulation and monitoring of clinical software systems by users, vendors, and regulatory agencies. Organizations assisting in development of recommendations, or endorsing the consortium position include the American Medical Informatics Association, the Computer-based Patient Record Institute, the Medical Library Association, the Association of Academic Health Sciences Libraries, the American Health Information Management Association, the American Nurses Association, the Center for Healthcare Information Management, and the American College of Physicians. The consortium proposes four categories of clinical system risks and four classes of measured monitoring and regulatory actions that can be applied strategically based on the level of risk in a given setting. The consortium recommends local oversight of clinical software systems, and adoption by healthcare information system developers of a code of good business practices. Budgetary and other constraints limit the type and number of systems that the FDA can regulate effectively. FDA regulation should exempt most clinical software systems and focus on those systems posing highest clinical risk, with limited opportunities for competent human intervention. PMID- 9391934 TI - Updating the Read Codes: user-interactive maintenance of a dynamic clinical vocabulary. AB - The Read Codes are a hierarchically-arranged controlled clinical vocabulary introduced in the early 1980s and now consisting of three maintained versions of differing complexity. The code sets are dynamic, and are updated quarterly in response to requests from users including clinicians in both primary and secondary care, software suppliers, and advice from a network of specialist healthcare professionals. The codes' continual evolution of content, both across and within versions, highlights tensions between different users and uses of coded clinical data. Internal processes, external interactions and new structural features implemented by the NHS Centre for Coding and Classification (NHSCCC) for user interactive maintenance of the Read Codes are described, and over 2000 items of user feedback episodes received over a 15-month period are analysed. PMID- 9391935 TI - Natural language generation in health care. AB - Good communication is vital in health care, both among health care professionals, and between health care professionals and their patients. And well-written documents, describing and/or explaining the information in structured databases may be easier to comprehend, more edifying, and even more convincing than the structured data, even when presented in tabular or graphic form. Documents may be automatically generated from structured data, using techniques from the field of natural language generation. These techniques are concerned with how the content, organization and language used in a document can be dynamically selected, depending on the audience and context. They have been used to generate health education materials, explanations and critiques in decision support systems, and medical reports and progress notes. PMID- 9391936 TI - Evaluating the coverage of controlled health data terminologies: report on the results of the NLM/AHCPR large scale vocabulary test. AB - OBJECTIVE: To determine the extent to which a combination of existing machine readable health terminologies cover the concepts and terms needed for a comprehensive controlled vocabulary for health information systems by carrying out a distributed national experiment using the Internet and the UMLS Knowledge Sources, lexical programs, and server. METHODS: Using a specially designed Web based interface to the UMLS Knowledge Source Server, participants searched the more than 30 vocabularies in the 1996 UMLS Metathesaurus and three planned additions to determine if concepts for which they desired controlled terminology were present or absent. For each term submitted, the interface presented a candidate exact match or a set of potential approximate matches from which the participant selected the most closely related concept. The interface captured a profile of the terms submitted by the participant and for each term searched, information about the concept (if any) selected by the participant. The term information was loaded into a database at NLM for review and analysis and was also available to be downloaded by the participant. A team of subject experts reviewed records to identify matches missed by participants and to correct any obvious errors in relationships. The editors of SNOMED International and the Read Codes were given a random sample of reviewed terms for which exact meaning matches were not found to identify exact matches that were missed or any valid combinations of concepts that were synonymous to input terms. The 1997 UMLS Metathesaurus was used in the semantic type and vocabulary source analysis because it included most of the three planned additions. RESULTS: Sixty-three participants submitted a total of 41,127 terms, which represented 32,679 normalized strings. More than 80% of the terms submitted were wanted for parts of the patient record related to the patient's condition. Following review, 58% of all submitted terms had exact meaning matches in the controlled vocabularies in the test, 41% had related concepts, and 1% were not found. Of the 28% of the terms which were narrower in meaning than a concept in the controlled vocabularies, 86% shared lexical items with the broader concept, but had additional modification. The percentage of exact meanings matches varied by specialty from 45% to 71%. Twenty-nine different vocabularies contained meanings for some of the 23,837 terms (a maximum of 12,707 discrete concepts) with exact meaning matches. Based on preliminary data and analysis, individual vocabularies contained < 1% to 63% of the terms and < 1% to 54% of the concepts. Only SNOMED International and the Read Codes had more than 60% of the terms and more than 50% of the concepts. CONCLUSIONS: The combination of existing controlled vocabularies included in the test represents the meanings of the majority of the terminology needed to record patient conditions, providing substantially more exact matches than any individual vocabulary in the set. From a technical and organizational perspective, the test was successful and should serve as a useful model, both for distributed input to the enhancement of controlled vocabularies and for other kinds of collaborative informatics research. PMID- 9391937 TI - Q-methodology: definition and application in health care informatics. AB - OBJECTIVE: To introduce the Q-methodology research technique to the field of health informatics. Q-methodology--the systematic study of subjectivity--was used to identify and categorize the opinions of primary care physicians and medical students that contributed to our understanding of their reasons for acceptance of and/or resistance to adapting information technologies in the health care workplace. DESIGN: Thirty-four physicians and 25 medical students from the Chicago area were surveyed and asked to rank-order 30 opinion statements about information technologies within the health care workplace. The Q-methodology research technique was employed to structure an opinion typology from their rank ordered statements. (The rank-ordered sorts were subjected to correlation and by person factor analysis to obtain groupings of participants who sorted the opinion statements into similar arrangements.) RESULTS: The typology for this study revealed groupings of similar opinion-types associated with the likelihood of physicians and medical students to adapt information technology into their health care workplace. A typology of six opinions was identified in the following groups: (1) Full-Range Adopters; (2) Skills-Concerned Adopters; (3) Technology Critical Adopters; (4) Independently-Minded and Concerned; (5) Inexperienced and Worried; and (6) Business-Minded and Adaptive. It is imperative to understand that in the application of Q-methodology, the domain is subjectivity and research is performed on small samples. The methodology is a combination of qualitative and quantitative research techniques that reveals dimensions of subjective phenomena from a perspective intrinsic to the individual to determine what is statistically different about the dimensions and to identify characteristics of individuals who share common viewpoints. Low response rates do not bias Q methodology because the primary purpose is to identify a typology, not to test the typology's proportional distribution within the larger population. CONCLUSION: Q-methodology can allow for the simultaneous study of objective and subjective issues to determine an individual's opinion and forecast their likeliness to adapt information technologies in the health care workplace. This study suggests that an organization's system implementers could employ Q methodology to individualize and customize their approach to understanding the personality complexities of physicians in their organization and their willingness to adapt and utilize information technologies within the workplace. PMID- 9391938 TI - Automated evidence-based critiquing of orders for abdominal radiographs: impact on utilization and appropriateness. AB - OBJECTIVE: Inappropriate utilization of diagnostic testing has been well documented. The purpose of this study was to measure the impact of presenting real time, evidence-based critiques about the appropriateness of abdominal radiograph (KUB) orders on physician decision making. DESIGN: Prospective trial where evidence-based critiques were presented to ordering clinicians in two kinds of situations: (1) a KUB was likely to have a low probability of providing useful information, or (2) an alternative view(s) was more appropriate given the clinical circumstance. There were two phases of the trial: Phase 1 was a 9-week period where evidence-based critiques were presented at the time of ordering a KUB, followed by Phase 2, a 19-week period in which orderers were randomized to receive critiques either amended to include both institutional data regarding the utility of the critiques and stronger messages about the lack of utility of the study, or the same critiques as presented in Phase 1, depending upon indication. Based upon the radiologist's report of their interpretation of the exams, the results of the examinations were scored as positive, equivocal, or negative using structured criteria. RESULTS: 299 KUBs in Phase 1 and 385 KUBs in Phase 2 received at least one critique. Cancellation rates of low yield films were low, and were similar in Phase 1 and 2, 8/258 (3%) vs. 10/283 (4%). Compliance with the recommendation for alternative view(s) was higher: 19/104 (38%) in Phase 1 vs. 96/176 (55%) in Phase 2 (p = 0.006). The results differentiated low-yield from non-low-yield films: 5% of low-yield films vs. 20% of non-low-yield films were positive in Phase 2 (p < 0.0001). Surgical physicians were less likely to cancel (p = 0.07) or to change to the suggested view(s) (p < 0.0001) than medical physicians or nurses. CONCLUSIONS: The intervention identified clinical situations in which KUBs appeared to have a low clinical yield. In response to evidence-based critiques, providers were reluctant to cancel their order, but were more willing to change to different views. To reduce the number of inappropriate radiographic films, stronger incentives or interventions may be required. PMID- 9391939 TI - Information technology in the community: the right tools for the job. PMID- 9391940 TI - Complex PTSD in victims exposed to sexual and physical abuse: results from the DSM-IV Field Trial for Posttraumatic Stress Disorder. AB - Two hundred thirty four participants in the DSM-IV Posttraumatic Stress Disorder (PTSD) Field Trial who reported sexual and/or physical abuse were evaluated. Participants were categorized according to type of abuse (physical, sexual, both), duration of abuse (acute versus chronic), and onset of abuse (early versus late). Separate logistic regression analyses examined the relationship between age of onset, duration, abuse type, and the complex PTSD (CP) lifetime diagnosis for women and men. Sexually abused women, especially those who also experienced physical abuse, had a higher risk of developing CP, although CP symptoms occurred at a high base rate among physically abused women. The theoretical implications and incremental clinical usefulness of targeting CP symptoms with abused populations are discussed. PMID- 9391941 TI - Adult memories of childhood trauma: a naturalistic clinical study. AB - The clinical evaluations of 77 adult psychiatric outpatients reporting memories of childhood trauma were reviewed. A majority of patients reported some degree of continuous recall. Roughly half (53%) said they had never forgotten the traumatic events. Two smaller groups described a mixture of continuous and delayed recall (17%) or a period of complete amnesia followed by delayed recall (16%). Patients with and without delayed recall did not differ significantly in the proportions reporting corroboration of their memories from other sources. Idiosyncratic, trauma-specific reminders and recent life crises were most commonly cited as precipitants to delayed recall. A previous psychotherapy was cited as a factor in a minority (28%) of cases. By contrast, intrusion of new memories after a period of amnesia was frequently cited as a factor leading to the decision to seek psychotherapy. The implications of these findings are discussed with respect to the role of psychotherapy in the process of recovering traumatic memories. PMID- 9391942 TI - Predicting PTSD in women with a history of childhood rape. AB - The impact of factors that predispose childhood rape victims to develop posttraumatic stress disorder (PTSD) is important in understanding both the impact of childhood rape and the development of PTSD as a psychological disorder. The present study attempted to determine which crime, perpetrator, victim, and aftermath characteristics are related to PTSD status. A national representative sample of women (N = 3,220) were interviewed about their history of rape, trauma related variables, and PTSD status. Consistent with research on crime victims, life threat and physical injury discriminated PTSD status in a sample of childhood rape victims. In addition, two other domains were related to PTSD development: (1) testification about rape and (2) rape types. The present findings are discussed in relation to previous research. PMID- 9391943 TI - The process in psychological debriefings. AB - Critical Incident Stress Debriefings have become an intervention method used in various cultures, countries and groups following critical incidents. Although the structure of such meetings has been adequately described, utilization of the group processes involved has received less attention. A model, process debriefing (PD), based on experiences from Europe, is presented. Some differences between the current CISD process in the United States and the Europe based model are outlined. Various factors that impact the process of debriefings are discussed with a special emphasis on leadership, and implications of these group process variables for psychological debriefing are presented. It is emphasized that the continued exploration and discussion of process issues is critical to advance the understanding of the critical elements of debriefing. PMID- 9391944 TI - Predictors of emotional numbing in posttraumatic stress disorder. AB - Little is known about the mechanisms underlying emotional numbing (EN). The functional relationship between other classes of posttraumatic stress disorder (PTSD) symptoms and EN is also not well understood. In the present study, we examined the statistical predictors of EN. We hypothesized that the severity of EN would be most strongly associated with the hyperarousal symptoms rather than the avoidance symptoms of PTSD, or comorbid depression or substance abuse. This prediction was derived from psychological and biological models that posit EN to be a product of the depletion of emotional resources subsequent to chronic hyperarousal. Using hierarchical multiple regression in two separate samples of Vietnam combat veterans, we found hyperarousal symptoms to be the most robust predictor of EN. These data suggest that there is a substantive relationship between hyperarousal symptoms and EN in PTSD. PMID- 9391946 TI - Attentional bias in posttraumatic stress disorder. AB - This study investigated preferential encoding of threat material in subjects with posttraumatic stress disorder (PTSD) with a modified dot-probe paradigm. This paradigm indexes attentional bias by measuring response latency to name neutral target words that are presented adjacent to or distant from threat words. Motor vehicle accident survivors with PTSD (n = 15), subclinical PTSD (n = 15), and low anxiety (n = 15) were required to name target words that were presented either adjacent to or distant from strong threat, mild threat, positive, and neutral words. PTSD subjects named targets faster when they were in close proximity to mild threat words. Results suggested that PTSD subjects' attention was drawn to the mild threat stimuli and are discussed in the context of network models of PTSD. PMID- 9391945 TI - Prolonged trauma and subsequent suicidal behavior: child abuse and combat trauma reviewed. AB - Stressful events have long been acknowledged as important risk factors for suicidal behavior. Although suicide research has generally focused on less severe stressful events, a long-standing vulnerability for suicidal behavior may be a sequela of prolonged traumatic stressors. The present paper discusses the relationship between prolonged traumatic stress and subsequent suicidality by reviewing studies that have examined suicidal behavior in relationship to child abuse and combat trauma. Traumatic stress is conceptualized according to a person environment interactional paradigm, and this paradigm is used to discuss the characteristics of traumatic events, recovery environments, and individuals that may contribute to subsequent suicidality. PMID- 9391947 TI - Replication and extension of a risk profile for Amerasian youth. AB - The relationship between number of risk factors and symptoms of anxiety and depression was examined in a cohort of Vietnamese Amerasians, replicating a study done with a previous cohort. One hundred forty seven subjects awaiting U.S. placement completed the Hopkins Symptom Checklist, the Vietnamese Depression Scale, and a questionnaire which included items found to be risk factors for psychological distress among Amerasians. Number of risk factors was linearly related to symptoms of both depression and anxiety. Results are consistent with previous findings of the relationship between risk factors and symptoms of psychological distress. The profile may be helpful in anticipating which refugees may be at risk for future psychological distress, and thus be useful in preventively allocating scarce treatment resources. PMID- 9391948 TI - Time-limited psychotherapy with Operation Desert Storm veterans. AB - Time-limited psychotherapy conducted 2 to 9 months after demobilization was evaluated with Persian Gulf Theater veterans of Operation Desert Storm (ODS). Thirty five treatment-seeking veterans were contrasted with 20 non-treatment seeking ODS Persian Gulf veterans in a repeated measures design at pretest, posttest, and 6-week followup assessments. In addition, psychotherapy participants at followup were contrasted with 80 non-treatment-seeking ODS Persian Gulf veterans from the same military units who were assessed one time at a comparable time point. Time-limited psychotherapy was associated with sustained improved psychosocial functioning and reduced levels of psychiatric and stress related symptomatology. PMID- 9391949 TI - An empirical evaluation of eye movement desensitization and reprocessing (EMDR) with survivors of a natural disaster. AB - Controlled studies of treatments effective with victims of natural disasters are almost nonexistent. This is a small study conducted under difficult conditions to test the effectiveness of Eye Movement Desensitization and Reprocessing (EMDR) in treating trauma related reactions following Hurricane Andrew. The results were positive in that EMDR produced significant improvement over wait list controls in perceived posttraumatic avoidance behaviors and thoughts as measured by changes in the Impact of Event Scale and significant improvement in subjective aversive reactions to representative experiences of the hurricane. These results suggest and support other studies that EMDR can be an effective therapeutic intervention for trauma reactions. PMID- 9391950 TI - Measurement of perceived disruption during rebuilding following Hurricane Andrew. AB - The purpose of this study was to develop and evaluate a measure of perceived disruption during rebuilding following a disaster. Two eight-item scales, which measured intensity of disruption during the entire repair phase (Intensity-RP) and intensity of disruption during the past month (Intensity-PM) were developed and administered to 135 survivors of Hurricane Andrew. At 9 to 12 months postdisaster, Intensity-RP and Intensity-PM were both significantly associated with scores on the Global Severity Index of the SCL-90-R, and with scores on the Impact of Event-Intrusion Scale; Intensity-PM alone was significantly associated with PTSD scores. Regression analyses indicated that each scale contributed significant unique variance in predicting mental health symptoms, even after controlling for relevant demographic and initial disaster exposure variables. PMID- 9391951 TI - Frequency and severity of panic attack symptoms in a treatment seeking sample of trauma victims. AB - This study assessed the frequency and severity of panic attack symptoms and panic attacks that develop in relation to the experience of traumatic events in 62 subjects seeking treatment for trauma-related symptomatology. Results indicated a high incidence of panic attacks (69%). Many individuals also thought they were going crazy or losing control (72%) or having a heart attack (38%) within the 2 weeks prior to assessment. These findings indicate that similar to panic disordered patients, many trauma victims with posttraumatic stress disorder (PTSD) not only experience physiological symptoms of panic, but are also fearful of these symptoms. PMID- 9391952 TI - Antidepressant treatment, posttraumatic stress disorder, survivor guilt, and spiritual awakening. AB - A patient with posttraumatic stress disorder (PTSD) had a major depressive episode that was responsive to treatment with the antidepressant fluoxetine. In contrast to the remission of other symptoms of depression, the associated feature of survivor guilt became more dramatically obvious. Individualized treatment of survivor guilt may be needed for patients with PTSD and major depression. PMID- 9391953 TI - Perspectives on the diagnosis, epizootiology, and control of the 1973 duck plague epizootic in wild waterfowl at Lake Andes, South Dakota. AB - An epizootic of duck plague occurred in early 1973 in a population of 163,500 wild waterfowl, primarily mallards (Anas platyrhynchos), wintering on Lake Andes and the nearby Missouri River in southeastern South Dakota (USA). The diagnosis was based on pathologic lesions and confirmed by virus isolation. Control measures included quarantine, attempts to reduce virus contamination of the area, dispersal of waterfowl, and monitoring of wild waterfowl populations for mortality. The epizootic resulted in documented mortality of 18% and estimated mortality of 26% of the waterfowl at risk. Prompt implementation of control measures might have limited mortality to approximately 8%. Losses during the epizootic were equivalent to 0.12% of the annual mortality in the North American 1996 fall population of 80,000,000 wild ducks. The most likely sources of the infection were free-flying wild mallard or American black duck (Anas rubripes) carriers from the upper midwestern or northeastern United States. Duck plague serum neutralization antibodies were demonstrated in 31% of 395 apparently healthy mallards sampled prior to dispersal of the flock at Lake Andes, suggesting that tens of thousands of potential duck plague carriers entered the wild waterfowl populations of all four major flyways. Consequently, the absence of major epizootics of duck plague in wild waterfowl in the subsequent two decades is evidence that substantial numbers of duck plague carriers can occur in wild waterfowl populations without resulting in epizootic mortalities. The failure to isolate duck plague virus from apparently healthy mallards sampled during the epizootic raises questions concerning the validity of conclusions regarding the status of duck plague in wild waterfowl based upon negative results of random surveys conducted in the absence of epizootics. PMID- 9391954 TI - Dynamics of plague in a Gunnison's prairie dog colony complex from New Mexico. AB - A plague (Yersinia pestis) epizootic spread through Gunnison's prairie dogs (Cynomys gunnisoni), and possibly other rodent species, in the Moreno Valley in north-central New Mexico between winter 1984-1985 and autumn 1987. We observed the progress of the epizootic and subsequent population recovery at four prairie dog towns within the valley during this period. At two towns (Midlake and Val Verde) the prairie dogs were marked prior to the epizootic. At two additional towns (Vega and South Entrance) prairie dogs were marked following the epizootic. In 1988, a second epizootic occurred at Vega. One hundred thirty-nine serum samples were collected from prairie dogs and other rodents and 1,750 fleas were collected from animals and burrows. Fleas infected with Y. pestis were collected from prairie dogs, deer mice (Peromyscus maniculatus), and thirteen-lined ground squirrels (Spermophilus tridecemlineatus). Prairie dog fleas included Oropsylla hirsuta, O. labis and O. tuberculata, deermouse associated fleas were Aetheca wagneri and Rhadinopsylla sectilis, and Oropsylla bacchi was associated with thirteen-lined ground squirrels. All of the above flea species were collected from prairie dog burrows. All rodent species shared some flea species. Thirteen lined ground squirrels disappeared shortly before plague was identified in prairie dogs at Midlake. Meadow voles were rare following the epizootic at Vega in 1986, became abundant in 1987, and disappeared at the time of the second prairie dog epizootic in summer 1988. Although we collected serum from Gunnison's prairie dogs, thirteen-lined ground squirrels, deer mice, and meadow voles (Microtus pennsylvanicus), we identified elevated serum titers against Y. pestis only in Gunnison's prairie dogs. Prairie dog mortality at all towns affected by plague was in excess of 99%. Serum antibody titers indicate that more than 40% of the few prairie dogs left to establish colonies following epizootics survived plague infection. PMID- 9391955 TI - Plague in a complex of white-tailed prairie dogs and associated small mammals in Wyoming. AB - Fleas were collected from white-tailed prairie dogs (Cynomys leucurus) and other small mammals trapped on six grids during a field study near Meeteetse (Wyoming, USA) in 1989 and 1990 to investigate the dynamics of plague in this rodent population. Fleas were identified and tested for Yersinia pestis by mouse inoculation. Yersinia pestis-positive fleas were found on prairie dogs and in their burrows. Flea species on prairie dogs changed from spring to late summer. White-tailed prairie dog numbers were significantly lower in the presence of Y. pestis-positive fleas; however, affected populations generally recovered 1 to 2 yr following absence of detectable plague. Grids where recovery occurred had a high proportion of juvenile male prairie dogs. Eighteen flea species were identified on small mammals, six of which were infected with Y. pestis. Some flea species were associated with a particular small mammal species, while others were found on a broad range of host species. Flea species most important in the potential interchange of Y. pestis between associated small mammals and white tailed prairie dogs were Oropsylla tuberculata cynomuris, Oropsylla idahoensis, and Oropsylla labis. Plague cycled through the white-tailed prairie dog complex in an unpredictable manner. Each summer the complex was a mixture of colonies variously impacted by plague: some were declining, some were unaffected by plague, and others were recovering from plague population declines. These data provide insight into the dynamics of plague in white-tailed prairie dog complexes, but predicting movement of plague is not yet possible and the role of associated mammals in maintenance of plague is not understood. PMID- 9391956 TI - Humoral immune response of cottontail rabbits naturally infected with Francisella tularensis in southern Illinois. AB - Cottontail rabbits (Sylvilagus floridanus) usually are thought to succumb to infection with Francisella tularensis. Reports of a rabbit population from southern Illinois (USA) with a high prevalence of F. tularensis antibodies suggested that some cottontails survived infection with this typically fatal bacterium. Our goal was to examine the humoral response of cottontails from a study area in southern Illinois for which multiple serum samples existed. Multiple sera were collected from 79 cottontails from 1986 to 1990 and 63% gained, lost, or maintained ELISA titers of IgM and IgG isotype antibodies. The typical pattern of antibody response appeared to be IgM isotype antibodies first, followed by IgG isotype antibodies, with both generally increasing to high titers. Negative culture attempts of liver tissue from 51 cottontails with varying antibody responses suggested that chronic infection did not occur in rabbits that developed antibody. The significance of the cottontail antibody response in resolution or prevention of tularemia infection remains unclear. PMID- 9391957 TI - Evaluation of a multivalent Pasteurella haemolytica vaccine in bighorn sheep: safety and serologic responses. AB - We examined effects of a multivalent Pasteurella haemolytica vaccine (serotypes A1, A2, T10) on humoral immune responses and P. haemolytica isolation rates in bighorn sheep (Ovis canadensis). Thirty captive bighorns, divided into groups of three on the basis of age, sex, and previous history of pneumonic pasteurellosis, received 0, 1, or 2 vaccine doses. Mild, transient lameness in most bighorns 1 day after initial vaccination was the only adverse effect observed. Oropharyngeal (> or = 75%) and nasal (< or = 50%) isolation rates for P. haemolytica did not differ among treatment groups. Ten of 36 distinguishable biogroup variants accounted for about 87% of the 464 P. haemolytica isolates from bighorns, but prevalences of specific biogroups were not affected by vaccination. Bighorns receiving 1 or 2 vaccine doses showed marked elevations in leukotoxin neutralizing antibody titers beginning 1 wk after vaccination. Agglutinating antibody titers to serotype A1 and A2 surface antigens were also elevated in vaccinated bighorns within 2 wk after vaccination; agglutinating antibody titers to serotype T10 surface antigens were relatively high in all three groups but appeared unaffected by vaccination. Vaccination 7 to 14 wk prior to parturition elevated leukotoxin neutralizing antibody titers in colostrum, but neither leukotoxin neutralizing nor serotype A1 surface antigen agglutinating antibody titers differed through 16 wk of age among lambs born to dams from different vaccine dose groups. Our data demonstrate that this multivalent P. haemolytica vaccine is safe and stimulates marked antibody responses in bighorn sheep. Further evaluation of this vaccine as a tool in preventing and managing pasteurellosis in bighorn sheep appears warranted. PMID- 9391958 TI - Bovine tuberculosis in free-ranging white-tailed deer from Michigan. AB - A 4.5 yr-old male white-tailed deer (Odocoileus virginianus) killed by a hunter during the 1994 firearm hunting season in northeastern Michigan (USA) had lesions suggestive of tuberculosis and was positive on culture for Mycobacterium bovis the causative agent for bovine tuberculosis. Subsequently, a survey of 354 hunter harvested white-tailed deer for tuberculosis was conducted in this area from 15 November 1995 through 5 January 1996. Heads and/or lungs from deer were examined grossly and microscopically for lesions suggestive of bovine tuberculosis. Gross lesions suggestive of tuberculosis were seen in 15 deer. Tissues from 16 deer had acid-fast bacilli on histological examination and in 12 cases mycobacterial isolates from lymph nodes and/or lungs were identified as M. bovis. In addition, lymph nodes from 12 deer (11 females and 1 male) without gross or microscopic lesions were pooled into 1 sample from which M. bovis was cultured. Although more male (9) than female (3) deer had bovine tuberculosis infections, this difference was not statistically significant. Mycobacterium bovis culture positive deer ranged in age from 1.5 to 5.5 yr with a mean of 2.7 yr (median 2.5 yr) for males and 3.2 yr (median 3.5 yr) for females. This appears to be the first epidemic occurrence of M. bovis in free-ranging cervids in North America. A combination of environmental (high deer density and poor quality habit) and management-related factors (extensive supplemental feeding) may be responsible for this epizootic. PMID- 9391959 TI - Upper respiratory tract disease and mycoplasmosis in desert tortoises from Nevada. AB - A population of desert tortoises (Gopherus agassizii) at Yucca Mountain (Nevada, USA) was monitored during four sampling periods using enzyme-linked immunosorbent assays (ELISA) to determine the percentage of individuals that had been exposed to Mycoplasma agassizii, a causative agent of upper respiratory tract disease. Respiratory tract disease has been considered a significant factor in the decline of desert tortoise populations in the Mojave Desert (USA). Few differences between sexes in ELISA values or percentages testing positive were noted. From 15 to 23% of samples per period tested positive for exposure to the mycoplasma. However, we noted few clinical signs of upper respiratory tract disease. This is in contrast to an earlier study which reported a similar proportion of seropositive tortoises as well as a high percentage of tortoises with clinical signs. However, our results are consistent with that study's conclusion that seropositivity for M. agassizii was a poor predictor of the likelihood to exhibit clinical signs of upper respiratory tract disease. Earlier reported epizootics of mycoplasma-associated respiratory disease occurred mainly during times of drought. Our samples were collected during a period of average to above-average rainfall, suggesting that manifestation of clinical signs of the disease may depend upon the physiological condition of tortoises which, in turn, is related to environmental conditions. PMID- 9391960 TI - Duration of Borrelia burgdorferi infectivity in white-footed mice for the tick vector Ixodes scapularis under laboratory and field conditions in Ontario. AB - The duration of Borrelia burgdorferi infectivity in white-footed mice (Peromyscus leucopus) experimentally inoculated or infested with infected Ixodes scapularis nymphs was evaluated. Infectivity was assessed by infesting these mice with unfed I. scapularis larvae at 7, 21, 35 and 49 days post-inoculation (DPI) or post infestation (PI). At 7 DPI, B. burgdorferi was transmitted from 18 of 24 syringe inoculated mice and all three tick-infected mice to I. scapularis larvae which fed upon them. However, at 21, 35 and 49 DPI, significantly fewer mice were infective. Borrelia burgdorferi was isolated from tissues of 14 of 22 syringe inoculated mice about 56 DPI, and from all three tick-infected mice. However, the level of agreement between xenodiagnosis and bacterial culture was no greater than would be expected by chance alone. We also determined if B. burgdorferi infectivity of mice varied in relation to periods of tick feeding in the field. White-footed mice were trapped during April, July and August 1993 from two habitats on Long Point peninsula (Ontario, Canada), where B. burgdorferi is endemic. Mice from each habitat were infested with laboratory-reared I. scapularis larvae. Ticks from each mouse were subsequently examined by immunofluorescent assay for B. burgdorferi infection and mice were cultured for B. burgdorferi. None of 3577 I. scapularis larvae fed on 62 mice captured within the cottonwood dune habitat were infected with B. burgdorferi, although it was isolated from six of these mice. Within the maple forest habitat, 0/24, 8/21 (38%) and 1/21 (5%) mice transmitted B. burgdorferi to I. scapularis larvae during April, July and August, respectively. Most mice from the maple forest with B. burgdorferi-positive tissues (14/21) were collected during July, although the level of agreement between xenodiagnosis and tissue culture was poor. Because B. burgdorferi infectivity in mice appears to be of short duration, overwintered I. scapularis larvae and nymphs may have to feed upon infected hosts at the same time of year in order for a cycle of B. burgdorferi infection to be maintained on Long Point. Infected I. scapularis nymphs, rather than persistently infected vertebrate hosts, likely serve as the overwintering "reservoir" for B. burgdorferi on Long Point. PMID- 9391961 TI - Vitreous humor analysis for selected biochemical parameters from cervids in Idaho. AB - Vitreous humor and liver samples were collected from hunter-harvested elk (Cervus elaphus) and mule deer (Odocoileus hemionus) in Idaho (USA). Concentrations of calcium, chloride, potassium, sodium, urea nitrogen and selenium were determined and evaluated according to species, age, gender, geographic location, and time elapsed following death. Vitreous humor analysis yielded reliable biochemical information for < or = 96 hr subsequent to the death of the animal. Vitreous potassium concentration changes over time could be used to estimate the time that elapsed following death. PMID- 9391963 TI - Effects of sex, age, capturing method, and season on serum chemistry values of brown bears in Croatia. AB - Sixty seven serum samples collected from 43 European brown bears (Ursus arctos) from Croatia were tested for < or = 31 serum chemistry parameters. Results were grouped and compared by bears origin (method of capture), sex, age, mass, and season sampled. Greatest differences were found between captive and free-living bears, and minor differences were found when sex, age, mass, or season of sampling were compared. Creatine kinase was significantly different among three categories of bears with the highest mean value of 924 IU/l in snare captured free-living bears compared to 67.8 IU/l in captive ones. Results of these tests provide reference values for European brown bears. PMID- 9391962 TI - Hematologic and serum biochemical reference intervals for Florida panthers. AB - Ninety-four blood samples were collected from 48 (29 males and 19 females) free ranging Florida panthers (Felis concolor coryi) captured in southern Florida (USA) from 1983 to 1994 for routine hematological and serum biochemical analysis. Florida panthers in the northern portion of their range had significantly higher red blood cell (mean +/- SD = 7.923 x 10(6) +/- 0.854 x 10(6)/microliter), hemoglobin (12.53 +/- 1.66 g/dl), and packed cell volume (36.97 +/- 4.27%) values compared to those of panthers localized in more southern parts of Florida (7.148 x 10(6) +/- 1.045 x 10(6)/microliter, 11.60 +/- 1.62 g/dl, and 34.82 +/- 5.99%, respectively). Adults had significantly higher mean serum total protein (7.50 +/- 0.59 g/dl) and packed cell volume (36.90 +/- 4.97%) values than juveniles (6.88 +/- 0.49 g/dl and 34.54 +/- 5.30%). However, mean serum albumin concentrations were significantly higher in juveniles (3.80 +/- 0.26 g/dl) when compared to adult values (3.58 +/- 0.26 g/dl). Mean serum calcium concentrations were significantly higher in juveniles (10.33 +/- 0.39 mg/dl) than in adults (9.66 +/- 0.45 mg/dl). Additionally, mean serum iron concentrations were significantly higher in those panthers of intergrade genetic stock compared to values in those of authentic genetic stock (105.6 +/- 72.1 micrograms/dl versus 59.3 +/- 19.7 micrograms/dl, respectively). PMID- 9391964 TI - Immobilization of wild collared anteaters with ketamine- and xylazine hydrochloride. AB - Collared anteaters (Tamandua tetradactyla) were immobilized for clinical procedures as part of a wildlife rescue during the filling of a hydroelectric dam (Petit Saut, French Guiana) from March 1994 to March 1995. Two doses of ketamine hydrochloride (KH) (group I mean +/- SD = 11.2 +/- 1.4 mg/kg, group II = 19.7 +/- 1.3 mg/kg) in combination with xylazine hydrochloride (XH) (1.0 +/- 0.1 mg/kg) were evaluated in seven and 10 collared anteaters, respectively. Induction time did not differ between the two groups. Immobilization time was significantly longer in group II than in group I (48.3 +/- 15.8 min and 35.0 +/- 9.5 min, respectively), without lengthening the recovery process. Adverse effects were not observed. The degree of anesthesia and the muscle relaxation were better in group II than in group I. Rectal temperature decreased in both groups and was significantly higher in group II than in group I. Heart rate was significantly higher in group II than in group I at 5 min post-injection and decreased in group II. No effects on respiratory rate were observed. We recommend the 20 mg/kg KH -1 mg/kg XH combination, especially for manipulations longer than 30 to 40 min and for minor surgery procedures. PMID- 9391965 TI - Experimental adenovirus hemorrhagic disease in yearling black-tailed deer. AB - An apparently novel adenovirus was associated with an epizootic of hemorrhagic disease that is believed to have killed thousands of mule deer (Odocoileus hemionus) in California (USA) during 1993-1994. A systemic vasculitis with pulmonary edema and hemorrhagic enteropathy or a localized vasculitis associated with necrotizing stomatitis/pharyngitis/glossitis or osteomyelitis of the jaw were common necropsy findings in animals that died during this epizootic. Six black-tailed yearling deer (O. hemionus columbianus) were inoculated with purified adenovirus isolated from a black-tailed fawn that died of acute adenovirus hemorrhagic disease during the epizootic. Three of six inoculated deer also received intramuscular injections of dexamethasone sodium phosphate every 3 days during the study. Eight days post-inoculation, one deer (without dexamethasone) developed bloody diarrhea and died. Necropsy and histopathologic findings were identical to lesions in free-ranging animals that died of the natural disease. Hemorrhagic enteropathy and pulmonary edema were the significant necropsy findings and there was microscopic vascular damage and endothelial intranuclear inclusion bodies in the vessels of the intestines and lungs. Adenovirus was identified in necrotic endothelial cells in the lungs by fluorescent antibody staining, immunohistochemistry and by transmission electron microscopy. Adenovirus was reisolated from tissues of the animal that died of experimental adenovirus hemorrhagic disease. Similar gross and microscopic lesions were absent in four of six adenovirus-inoculated deer and in the negative control animal which were necropsied at variable intervals during the 14 wk study. One deer was inoculated with purified adenovirus a second time, 12 wk after the first inoculation. Fifteen days after the second inoculation, this deer developed severe ulceration of the tongue, pharynx and rumen and necrotizing osteomyelitis of the mandible which was associated with vasculitis and thrombosis of adjacent large vessels and endothelial intranuclear inclusions. Transmission electron microscopy demonstrated adenovirus within the nuclei of vascular cells and immunohistochemistry demonstrated adenovirus antigen within tonsilar epithelium and in rare vessels. PMID- 9391966 TI - Antibodies against equine herpesviruses in free-ranging mountain zebras from Namibia. AB - Twenty-one blood samples of free-ranging mountain zebras (Equus zebra) from Namibia were tested for equine herpesvirus (EHV-1, -2, -3, -4) specific antibodies by immunofluorescence assay (IFA) and neutralization test (NT). Additionally, type-specific nested polymerase chain reactions (nested PCR) were employed for detection of EHV-1, -2 and -4 DNA. Equine herpesvirus-1 antibodies were detected by IFA in all zebras, while only seven serum samples contained EHV 4 IFA antibodies. Sera with high IFA antibodies also were found to neutralize EHV 1 and -4. Furthermore, 20 zebras were EHV-2 seropositive by IFA, and one zebra had EHV-2 neutralizing antibodies. Equine herpesvirus-3 specific antibodies were not detected. We did not amplify EHV-1, -2 or -4 specific DNA sequences in peripheral blood leukocytes of the same zebras using type-specific nested PCR. EHV infections appear to be widespread in free-ranging zebras, as they are in domestic horses. PMID- 9391967 TI - A small-scale survey of hantavirus in mammals from Indiana. AB - In order to determine if hantaviruses were present in mice and other small mammals in Indiana (USA), small mammals were trapped in Brown, LaPorte, Tippecanoe and Whitley counties. Sixty-seven small mammals were trapped during August and September 1994. Sixty-three Peromyscus leucopus, one Microtus pennsylvanicus, one Zapus hudsonius and two Blarina brevicauda were captured and tested for hantaviruses. Six P. leucopus were found to have antibody to Sin Nombre virus (SN) by IgG ELISA, and a 139 bp fragment of SN-like hantavirus was amplified from five of them. All six of the positive P. leucopus were from LaPorte County. No other small mammals had evidence of infection with SN virus. This study presents the first report of Sin Nombre-like hantavirus in P. leucopus from Indiana. PMID- 9391968 TI - Serosurvey for selected viral diseases and demography of African wild dogs in Tanzania. AB - African wild dogs (Lycaon pictus) are endangered, with only 3,000-5,000 remaining in the wild. It is believed that wild dogs are unusually vulnerable to viral diseases, particularly rabies and canine distemper (CDV). However, canine distemper has been confirmed by laboratory diagnosis in only one free-living wild dog. The 43,000 km2 Selous Game Reserve (SGR; Tanzania) holds approximately 900 adult wild dogs. In a study area of 2,600 km2, the population maintained high density (> or = 1 dog/20.5 km2) from 1991 to 1996. The population was stable, varying 18% below and 9% above the mean density over the 6-yr period. Serum samples (n = 22) collected over 3 yr showed that most individuals were exposed to CDV (59%:95% confidence interval = 43-76% seropositive) and canine parvovirus (68%:95% CI = 54-81% seropositive), although none were seropositive for rabies (0%:95% CI = 0-17%). CDV titers were positively related to age, with no seropositive dogs younger than 1.9 yr. At least five of 13 dogs positive for CDV seroconverted during the study. Dogs with high CDV titers did not survive better in the years after sampling (mean survival +/- SE for those that died = 638 +/- 92 days,). Variation in mean litter size was inversely related to CPV exposure in the SGR and elsewhere. Annual mortality rates were low in comparison to other populations for all age classes (pups: 31 +/- 8%, n = 127, yearlings: 22 +/- 10%, n = 93, adults: 20 +/- 6%, n = 235). Annual mortality rates fluctuated little between 1992 and 1996. These data show that wild dog populations, like those of other canids, can remain stable and demographically healthy despite exposure to CDV and CPV. PMID- 9391969 TI - Winter mortality of common loons in Florida coastal waters. AB - Diagnostic findings are presented for 434 common loons (Gavia immer) found sick or dead on Florida beaches from 1970 through 1994, primarily during the months of December to April. The most commonly recognized problem was an emaciation syndrome (66%), followed by oiling (18%), aspergillosis (7%), trauma (5%) and miscellaneous disease entities (1%). The cause-of-death for 3% of the birds was not determined. Many of the carcasses examined (n = 173) were obtained during an epizootic which occurred from January to March of 1983 in which more than 13,000 loons were estimated to have died. An emaciation syndrome, characterized by severe atrophy of pectoral muscles, loss of body fat and hemorrhagic enteritis, was the primary finding in this epizootic. It was postulated to have a complex etiologic basis involving synergistic effects and energy costs of migration, molting and replacement of flight feathers, food resource changes, salt-loading, intestinal parasitism, environmental contaminants, and inclement weather. PMID- 9391970 TI - Uncinariasis in northern fur seal and California sea lion pups from California. AB - Northern fur seal (Callorhinus ursinus) (n = 25) and California sea lion (Zalophus californianus) (n = 53) pups, found dead on rookeries on San Miguel Island (California, USA), were examined for adult Uncinaria spp. Prevalence of these nematodes was 96% in fur seal pups and 100% in sea lion pups. Mean intensity of Uncinaria spp. per infected pup was 643 in fur seals and 1,284 in sea lions. Eggs of Uncinaria spp. from dead sea lion pups underwent embryonation in an incubator; development to the free-living third stage larva occurred within the egg. This study provided some specific information on hookworm infections in northern fur seal and California sea lion pups on San Miguel Island. High prevalence rate of Uncinaria spp. in both species of pinnipeds was documented and much higher numbers (2X) of hookworms were present in sea lion than fur seal pups. PMID- 9391971 TI - Ultrastructure of the cyst wall of Sarcocystis sp. in roe deer. AB - Samples of heart, tongue, oesophagus and diaphragm muscle from twenty-two naturally infected roe deer (Capreolus capreolus) harvested in central Italy were examined for sarcosporidiasis. The structure of Sarcocystis spp. muscle cysts was examined by light and electron microscopy. Only one type of thin-walled cyst was distinguished by light microscopy. Electron microscopy showed cysts having a thin highly folded primary cyst wall, without fibrillar material, that formed thin hair-like protrusions often having a T-form, especially close to host cell mithocondria. The cysts appeared to belong to a single Sarcocystis sp. so that all the animals had monospecific infections. This cyst was compared with cysts described in other cervid in an attempt to determine if single or multiple species of the genus Sarcocystis occur in the Cervidae. Apparently, a single Sarcocystis sp. with a low specificity for the intermediate host can infect the Cervidae. PMID- 9391972 TI - Sarcosporidiasis in rodents from Thailand. AB - One to six Sarcocystis spp. were identified in the skeletal muscles of 41 (33%) of 124 wild rodents (Rattus spp. and Bandicota indica) mainly captured in the central plains of Thailand throughout the year in 1995. Included were S. singaporensis, S. villivillosi, and S. murinotechis-like cysts all of which showed a striated cyst wall at the light microscopical level, and Sarcocystis cymruensis, S. sulawesiensis, and S. zamani which possessed smooth cyst walls. The ultrastructure of the cyst wall and other morphological characteristics used to distinguish species are described. By inoculation of muscle cysts from wild caught rodents into coccidia-free pythons (Python reticulatus, P. molurus bivittatus), we confirmed that P. reticulatus is a suitable definitive host for S. singaporensis and S. zamani in Thailand. Furthermore, we showed by fecal examination of reticulated pythons collected in the wild and subsequent experimental infection of laboratory rats that these hosts also are naturally infected with both species. Sarcocystis cymruensis is reported for the first time from Southeast Asia. This parasite was prevalent in brown rats (Rattus norvegicus) and bandicoot rats (B. indica) which were captured near human habitations; it is likely to be transmitted to rats via cats. The definitive hosts of S. sulawesiensis and S. murinotechis are unknown. Hence, at least three Sarcocystis spp. (S. singaporensis, S. zamani, S. villivillosi) are likely to cycle between snakes and rodents in agricultural areas in Thailand. Among these, S. singaporensis appears to be the most prevalent species. PMID- 9391973 TI - Extracting Protostrongylus spp. larvae from bighorn sheep feces. AB - First-stage larvae of Protostrongylus spp. were more numerous in the core of bighorn sheep (Ovis canadiensis canadiensis) pellets than near the surface. As a result, only 22% could be extracted from whole pellets and the numbers collected did not reflect the total number of larvae present in samples. Crushing semi dried pellets yielded seven times as many larvae and numbers collected were correlated with totals present. The use of tissue, in addition to a screen filter in a beaker extraction method, produced a cleaner sample and did not affect larval collection or the correlation. By comparison, most first-stage larvae of Parelaphostrongylus tenuis from white-tailed deer (Odocoileus virginianus) were near the surface of fecal pellets where they may be removed readily by water. PMID- 9391974 TI - Filarial dermatitis in a striped skunk. AB - A striped skunk (Mephitis mephitis) from Kansas (USA) with severe diffuse dermatitis characterized by extensive alopecic areas, thickened skin, and multiple, scattered cutaneous abscesses on the dorsal aspect of the head, neck, and trunk was submitted for diagnostic evaluation. More than 50 nematodes identified as Filaria taxideae were found in the dorsal subcutaneous tissue. Histologic examination of the skin revealed multifocal pyogranulomatous inflammation with intralesional larvated nematode eggs, moderate orthokeratotic hyperkeratosis, and mild acanthosis. The lesions resemble those reported from badgers (Taxidea taxus) and a lesser panda (Ailurus fulgens) with dermatitis caused by Filaria taxideae. Although F. taxideae has been previously collected from skunks, this is the first report of filarid dermatitis caused by this nematode in a striped skunk. PMID- 9391975 TI - Gnathostomiasis in frog-eating snakes from Japan. AB - Gnathostoma doloresi parasitizes the gastric wall of wild (boars) and domestic (pigs) swine (Sus scrofa). Its larvae cause cutaneous larva migrans in humans. Amphibians, reptiles and a freshwater fish are infected with the advanced 3rd stage larvae. Prevalence of G. doloresi larvae were surveyed in several snakes, especially in a common frog-eating snake (Rhabdophis tigrinus). All species of snakes examined were infected with G. doloresi larvae suggesting that snakes are important reservoir hosts. Prevalence of G. doloresi larvae in frog-eating snakes was lower than that found in mammal-eating snakes. Thus, as a source of infection to snakes, small mammals may be more important than frogs in the natural life cycle of G. doloresi in Japan. PMID- 9391976 TI - Capillariasis in the trachea of a raccoon. AB - Cross-sections of nematodes were seen in histologic sections of trachea from a raccoon (Procyon lotor) collected in Virginia (USA); they occupied epithelium and contained unembryonated, bioperculated eggs characteristic of the genus Capillaria (= Eucoleus). A mild inflammatory cell infiltrate in the lamina propria subjacent to the nematode was the only apparent host response. This is the first report of capillariasis in the trachea of raccoons. PMID- 9391977 TI - Fascioloidiasis in game-ranched elk from Montana. AB - The distribution of Fascioloides magna in game-ranched elk and the potential for spread of the parasite through movement of infected animals was examined in Montana (USA). Fecal samples (n = 448) collected from captive elk on 29 game ranches were examined for eggs of F. magna by fecal sedimentation. Eggs were detected in elk on 5 ranches. This suggests that F. magna has been translocated by infected game-ranched elk. The wide distribution of snail intermediate hosts for F. magna in Montana indicates a potential to spread the parasite to other captive cervids domestic livestock or free-ranging wildlife. PMID- 9391978 TI - A trematode metacercaria causing gill cartilage proliferation in steelhead trout from Oregon. AB - Gills of steelhead trout (Oncorhynchus mykiss) held in liveboxes to detect the presence of pathogens in the Willamette River (Oregon, USA) became heavily infected with trematode metacercariae. The metacercariae encysted adjacent to the cartilaginous rods of gill filaments and elicited a host response of cartilage proliferation from the perichondrium. Although some hyperplasia of gill epithelium and fusion of lamellae was apparent, the extent of damage to the respiratory surface was apparently insufficient to cause trout mortality. Morphological characteristics of the metacercariae did not allow precise identification, but they suggested affinities to either the Heterophyidae or Cryptogonimidae. Some heterophyids are known to cause proliferation of cartilage in fish gills, while cryptogonimids are not. This is the first report of trematode induced gill cartilage proliferation in steelhead trout. PMID- 9391979 TI - Tetraphyllidean cysticerci in the peritoneal cavity of the common dolphin. AB - Cysticerci of the cestodes Monorygma grimaldii and Phyllobothrium delphini were encountered during necropsy of an adult common dolphin (Delphinus delphis) found dead on the southeastern coast of Australia. Monorygma grimaldii cysticerci were found within highly organized retroperitoneal cysts, whereas P. delphini cysticerci in the subcutaneous blubber did not occupy specialized structures. There was a localized lymphoplasmacytic host response to the presence of cysticerci of both species, but M. grimaldii provoked a more severe suppurative response than P. delphini. The systematics and life history of both parasites are incompletely known, but sharks postulated as the potential definitive hosts are found in the region. A unique cysticercus of M. grimaldii was found lying free in the peritoneal cavity of this dolphin. Two rare records of M. grimaldii cysticerci in pinnipeds from the literature include one case of aberrant migration to the testis. PMID- 9391980 TI - Toxoplasmosis in naturally infected deer from Brazil. AB - Serum samples from 107 cervids were examined for Toxoplasma gondii antibodies using indirect hemagglutination (IHA), indirect immunofluorescence (IFA), enzyme linked immunosorbent assay (ELISA) and Dot-ELISA. Samples were obtained from 66 marsh deer (Blastocerus dichotomus) in the State of Sao Paulo (Brazil) and from 41 pampas deer (Ozotocerus bezoarticus) in the State of Goias (Brazil). Antibodies to T. gondii were found in 23 (22%) of the deer, with 18 and 5 positive samples, respectively, for B. dichotomus and O. bezoarticus. The highest prevalence of T. gondii antibodies were young adults (32%), following by adults (27%) and fawns (13%). Only one serum sample (8%) from a newborn fawn was positive in the serological tests. The convenience of the Dot-ELISA test is obvious when compared with other serological tests for both laboratory or field surveys, mainly due to its features of practicability and reagent stability. PMID- 9391981 TI - Tick paralysis in a red wolf. AB - A free-ranging male red wolf (Canis rufus) in North Carolina (USA), exhibiting paresis, anorexia and heavy tick infection was diagnosed with tick paralysis. The wolf recovered completely following the removal of all ticks. This is the first record of tick paralysis in the red wolf. PMID- 9391982 TI - Mucormycosis in a free-ranging green tree frog from Australia. AB - Mucor amphibiorum is reported for the first time from a free-ranging native amphibian, a green tree frog (Litoria caerulea) from Queensland, Australia. The nasal cavity was largely replaced by granulomatous inflammatory tissue, and most internal organs had nodular granulomas. Typical mother and daughter sphaerules of M. amphibiorum occurred in these nodules which were due to granulomatous inflammation as well as areas of more active mixed inflammation with necrosis. Tissue homogenate from the spleen was inoculated into two cane toads (Bufo marinus), and one toad became infected with M. amphibiorum. PMID- 9391983 TI - Salmonella arizonae sepsis in a lynx. AB - A 4.5-wk-old lynx (Felis lynx) was presented for necropsy with a history of poor growth, mild diarrhea, anemia, and lethargy. The liver was enlarged and had a 7 mm long fracture that resulted in severe intraabdominal hemorrhage and death. Microscopic lesions were indicative of severe ulcerative cystitis and septicemia. Pure cultures of Salmonella arizonae were isolated from the liver, kidney, and spleen. Based on differences in the chronicity of inflammation in the urinary bladder versus other organs, we speculate that chronic cystitis caused by S arizonae lead to septicemic infection. PMID- 9391984 TI - Rabies virus in the decomposed brain of an Ethiopian wolf detected by nested reverse transcription-polymerase chain reaction. AB - Approximately 75 individuals from a population of 111 Ethiopian wolves (Canis simensis) died or disappeared from the Bale Mountains National Park (Ethiopia) between 1988 and 1992 during two significant population declines. Confirmation of rabies virus in two carcasses was based on the fluorescent antibody test (FAT) and the mouse inoculation test (MIT). In an Ethiopian wolf brain previously designated rabies negative by both FAT and MIT, rabies virus was identified by nested reverse transcription-polymerase chain reaction (RT-PCR) and confirmed by Southern blot hybridization. These methods were successfully used on a highly decomposed brain sample which had been stored in 20% dimethyl sulfoxide. This test system allows early and sensitive detection to be undertaken to more effectively prevent spread of disease and thus protect surviving animals. PMID- 9391985 TI - Pseudorabies in captive coyotes. AB - Pseudorabies (Aujeszky's disease) was diagnosed in three adult captive coyotes (Canis latrans) from southern Indiana (USA). The coyotes died in their outdoor enclosure within a 48 hr period. Histopathology revealed multifocal, nonsuppurative meningioencephalitis and eosinophilic intranuclear inclusion bodies within neurons. Samples of brain were positive for pseudorabies virus by fluorescent antibody testing and virus isolation. Source of infection was the probable consumption of pseudorabies virus-infected pig carcasses. PMID- 9391986 TI - Serological evidence of bovine herpesviruses 1 and 2 in Asian elephants. AB - Antibodies were detected against bovine herpesviruses 1 (BHV 1) and 2 (BHV 2) in Asian elephants (Elephas maximus) using the passive hemagglutination (PHA) test. The study was conducted during May to December 1994 using sera collected from zoological gardens and national parks in India. Four (4%) of 109 elephant sera had PHA titers ranging from 1:8 to 1:32 against BHV 1. Twenty-five (23%) of the 109 elephant sera had PHA titers ranging from 1:8 to 1:64 against BHV 2. Asian elephants appear to be better reservoirs for herpesviruses which are serologically related to BHV 2. PMID- 9391987 TI - A stingray spine in the scapula of a bottlenose dolphin. AB - A stingray spine was found lodged in the scapula of a deceased 272 cm, male bottlenose dolphin (Tursiops truncatus) from South Carolina (USA) following skeletal preparation, nearly 6 mo after necropsy. No external puncture wound, internal bruising, or laceration of muscle tissue surrounding the scapula was evident during necropsy of the animal. Implantation of the spine did not appear to be related to the death of the dolphin, but probably occurred at an early age. Abnormal development of bone surrounding the spine resulted in the formation of a cavity at the wound site. Two mechanisms were considered as contributors for the cavity formation. These were the mechanical action of the spine stimulating the body's defense system for managing foreign objects, and the release of potent toxins from the spine sheath. PMID- 9391988 TI - Epidermal tumors of rainbow smelt with associated virus. AB - Epithelial tumors of the skin occurred in landlocked populations of rainbow smelt (Osmerus mordax) in several lakes in New Hampshire (USA) during the spawning runs. Histologically, these were noninvasive epithelial cell lesions. Herpesvirus like particles could be seen in the nucleus and cytoplasm. The lesions occurred in both males and females. Prevalence which varied annually, was as high as 30%. PMID- 9391989 TI - Psychosocial factors associated with the use of hormonal replacement therapy in a longitudinal follow-up of Swedish women. AB - OBJECTIVES: To follow up a cohort of 1400 women aged 52 years who had replied to a health questionnaire 4 years previously. The follow-up covered general and gynecological health, experience of symptoms, the use of hormone replacement, the reasons for starting HRT and effectiveness of treatment as well as comparison of users and nonusers concerning psychosocial factors and life style. METHODS: A questionnaire together with a letter was mailed to the women who had responded previously. The questionnaire covered four different areas: sociodemographic background, general and gynecological health, a 20-item symptom rating scale, and questions concerning work role. RESULTS: A total of 1194 women (85%) responded to the questionnaire; 40% of the women were currently using hormone replacement. The reasons for starting treatment were: relief of somatic (55%) and psychological symptoms (20%), increased wellbeing (5%), to prevent disease (5%) and other reasons, such as keeping young (15%). Positive effects were experienced by 86% and negative effects by 26%. Women using HRT had less frequent vasomotor symptoms, sleep problems and vaginal dryness and were more harmonious than nonusers. There were no differences between HRT users and nonusers regarding negative mood and sexual desire. Women with psychologically demanding and stressful jobs requiring concentration were more likely to use hormone replacement. CONCLUSIONS: Swedish women are increasingly willing to start hormone replacement, particularly those who suffer from vasomotor symptoms and who have stressful and psychologically demanding occupations. The majority of these experience relief of symptoms. A certain proportion will suffer from side effects and are likely to discontinue treatment. PMID- 9391991 TI - Menorrhagia--a search for epidemiological risk markers. AB - OBJECTIVE: To isolate epidemiological risk factors for menorrhagia. METHODS: Menstrual blood loss (MBL) of one bleeding episode of 182 healthy women was measured with the alkaline hematin method and the results were related to age, parity, body mass index and smoking habits. Multiple and logistic regression analysis was performed to isolate the variables that most influence MBL. Two consecutive menstrual episodes were measured in 117 women, to determine individual constancy. RESULTS: MBL increased significantly with age (Kruskal Wallis, P < 0.03) and the percentage of women with menorrhagia was significantly higher above 40 years of age (Mann-Whitney's ranks sum test, P < 0.05). The odds ratio of parous:nulliparous women for menorrhagia was 2.27:1, but after adjustment for age this influence disappeared. Body mass index and smoking habits were not significantly related to menorrhagia. The mean difference between the MBL of two consecutive menstruations is 2.1 ml (S.E.: 1.7, 95% CI: -1.3 to 5.5 ml). CONCLUSIONS: Only age could be indicated as a risk marker for menorrhagia. Parity, body mass index and smoking habits appear to have no significant effect on MBL, when adjusted for age. The individual constancy in MBL between two consecutive cycles is very high and therefore one single measurement suffices in studies of MBL. PMID- 9391990 TI - Comparison of the long-term effects of oral estriol with the effects of conjugated estrogen, 1-alpha-hydroxyvitamin D3 and calcium lactate on vertebral bone loss in early menopausal women. AB - We investigated the long-term effects of oral estriol (E3) on bone mineral density (BMD) at the lumbar spine and biochemical indices of bone turnover in early menopausal women. We studied 64 healthy early menopausal women who were treated for 24 months with 2.0 mg E3 plus 2.5 mg medroxyprogesterone acetate daily (E3 group, n = 15), 0.625 mg of conjugated estrogen plus 2.5 mg medroxyprogesterone acetate daily (CE group, n = 19), 1.0 microgram 1-alpha hydroxyvitamin D3 daily (D3 group, n = 13), or 1.8 g calcium lactate containing 250 mg of elemental calcium daily (Ca group, n = 17). The BMD at the third lumbar vertebra was determined by quantitative computed tomography, and serum levels of osteocalcin (OC) and total alkaline phosphatase (Alp), as well as urinary ratios of calcium-to-creatinine (Ca/Cr) and hydroxyproline-to-creatine (Hyp/Cr), were evaluated at baseline and every 6 months. After 24 months of treatment, the BMD decreased significantly by 12 +/- 4.5% (mean +/- S.E.) in the D3 group and 14 +/- 2.5% in the Ca group, but not in the E3 group (-4.1 +/- 4.8% from baseline) and in the CE group (-0.9 +/- 3.2% from baseline). The serum levels of Alp and OC decreased or remained unchanged in the E3 and CE groups, but increased in the D3 and Ca groups. The urinary Ca/Cr was decreased in the E3 and CE groups, but not in the D3 and Ca groups. The urinary Hyp/Cr decreased in the CE group, was unchanged in the E3 and D3 groups, and increased in the Ca group. Uterine bleeding occurred less frequently in the E3 than in the CE group (2.4 +/- 4.2 versus 13.1 +/- 14.8 days/person per year, P < 0.001). The bone-preserving effect of 2.0 mg of oral E3 was comparable to that of 0.625 mg of conjugated estrogen and was superior to that of 1.0 microgram 1-alpha-hydroxyvitamin D3 and 1.8 g Ca. Our findings suggest that a reduction in bone turnover in the E3 group may have contributed to the preservation of bone. PMID- 9391993 TI - Menopause in normal and uncomplicated NIDDM women: physical and emotional symptoms and hormone profile. AB - OBJECTIVE: To compare the physical characteristics, emotional symptoms and metabolic conditions of menopausal women with and without non insulin dependent diabetes mellitus (NIDDM). METHODS: We studied 100 menopausal women 45-72 years of age, 51 with and 49 without NIDDM, in a cross-sectional design. Biological characteristics were collected and emotional symptoms were assessed with a modified Hamilton and Bech-Rafaelsen scale, scoring depression, anxiety, non specific symptoms of depression (NSSD) and the empty nest syndrome (ENS). Weight, body mass index (BMI), waist/hip and abdomen/hip ratios and percent of body fat were registered. The sulfoconjugated form of the dehydroepiandrosterone (DHEAS), follicle stimulating hormone (FSH), cortisol and fasting, as well as postprandial insulin/glucose ratios, were measured in blood. RESULTS: Women with NIDDM had earlier mean age for menopause, more central obesity and less peripheral fat; they had also more prevalent emotional symptoms than non diabetic menopausal women. In women with NIDDM, symptoms were associated with years since diagnosis and with BMI. In non diabetic menopausal women schooling and attitudes to sexuality were associated with symptoms. FSH was inversely associated with BMI in both diabetic and non diabetic women; postprandial insulin/glucose ratio was correlated with central obesity in the group without NIDDM and cortisol with sitting systolic blood pressure (SBP) in the group with NIDDM. CONCLUSION: The diagnosis of NIDDM and its metabolic conditions were associated with an increased frequency of some symptoms in menopausal women. PMID- 9391992 TI - Modulatory effects of a synthetic steroid (tibolone) and estradiol on spontaneous and GH-RH-induced GH secretion in postmenopausal women. AB - OBJECTIVE: Since hormonal replacement therapy (HRT) affects plasma GH levels, the present study aimed to verify the effect of tibolone, a synthetic steroid, on modulating spontaneous and growth hormone releasing hormone (GH-RH) induced GH secretion. METHODS: Postmenopausal women (n = 30) were enrolled and randomly subdivided in three groups (n = 10 each group): (1) treated with transdermal estradiol (50 micrograms) (Dermestrill, Rottapharm, Monza, Italy) biweekly; (2) treated with transdermal estradiol (100 micrograms) (Dermestrill, Rottapharm, Monza, Italy) biweekly; (3) treated with tibolone 2.5 mg/day (Livial, Organon Italia, Rome, Italy). Patients underwent a GH-RH test (1 microgram/kg) and 15 of them underwent to a pulsatility study before and 5 weeks after treatment. RESULTS: Mean (+ S.E.M.) GH plasma levels increased in all patients after any type of HRT. GH response to GH-RH stimulation (expressed as maximal response to GH-RH or as delta value) was similar in the three groups while significant changes occurred in spontaneous pulsatile GH release. Tibolone and both dosages of transdermal estradiol significantly reduced GH pulse frequency and increased pulse amplitude. CONCLUSIONS: The reduced plasma GH levels observed during postmenopause are probably related to a reduced endogenous GH-RH and not to a reduced pituitary ability to respond to GH-RH. In addition tibolone, as well as transdermal estradiol, are effective in restoring the spontaneous GH episodic release. PMID- 9391994 TI - The effect of postmenopause and postmenopausal HRT on measured voice values and vocal symptoms. AB - OBJECTIVES: To study the effect of postmenopause and postmenopausal hormone replacement therapy (HRT) on the measured fundamental frequency (F0) and sound pressure level (SPL) of sustained phonation and speaking voice samples and on subjective vocal/laryngeal symptoms. METHODS: Forty-three postmenopausal women (mean age 51.6) were divided into three groups: a group with no HRT, an estrogen group (daily oral dose of 2 mg of estradiol valerate), and an estrogen-progestin group (daily oral dose of 2 mg of 17-B-estradiol and 1 mg of northisterone acetate). Voice measurements were made before and after 1 year of treatment. Subjective symptoms were registered using a questionnaire. RESULTS: The mean F0 and SPL decreased significantly in the group with no HRT in spontaneous speech and reading samples as did SPL in the normal phonation sample. In both groups with HRT, the mean F0 decreased significantly only in the spontaneous speech sample and the decrease was smaller than in the group with no HRT. The mean SPL decrease in the estrogen group was significant in the normal phonation sample while in the estrogen-progestin group it was significant in both the normal phonation and the reading sample. The number of subjective symptoms was smallest in the estrogen group. CONCLUSIONS: The changes in the measured voice values and the subjective symptoms experienced suggest that at least the early postmenopausal years are associated with vocal changes and that HRT counteracts this phenomenon. This seems to be more pronounced with estrogen than with a combination of estrogen and progestin. PMID- 9391995 TI - Hormone replacement therapy and intraocular pressure. AB - OBJECTIVES: To evaluate the effect of hormone replacement therapy (HRT) on intraocular pressure (IOP) in menopausal women. METHODS: The IOP of 25 white menopausal women without an abnormal ophthalmologic history was measured before and during HRT regimen. IOP fluctations were recorded before and 1, 4, and 12 weeks after the beginning of HRT. These measurements were obtained according to a standardized time schedule (08:00, 12:00, 16:00, and 19:00 h). RESULTS: The mean IOP in the left eye decreased from 16.2 +/- 2.4 mmHg before therapy to 14.0 +/- 2.1 mmHg after 12 weeks of therapy (P < 0.001). In the right eye, whose IOP was at 15.3 +/- 2.3 mmHg before therapy there was a decrease to 14.0 +/- 1.9 mmHg after 12 weeks of therapy (P < 0.001). CONCLUSION: Hormone replacement therapy has a positive effect on IOP in menopausal women. PMID- 9391996 TI - Decrease of bone formation in adult women with fragility fractures. AB - OBJECTIVES: To compare bone mineral density (BMD) and some markers of bone metabolism in women with fragility fractures and in normal age-matched subjects. METHODS: A 100 women with at least one vertebral deformity > 25%, and 219 age-, BMI- and parity-matched healthy women, were recruited for the study. In all the patients fractures were symptomatic and occurred at least 1 year before densitometric measurement. Forearm bone mineral density (BMD) as well as biochemical assessment of some markers of bone turnover were measured in all the subjects. RESULTS: BMD was significantly lower in the fracture than in the control group (0.326 +/- 0.073 vs. 0.379 +/- 0.079; P < 0.001). Fractured women showed alkaline phosphatase (ALP) and osteocalcin (OC) serum levels significantly lower than controls, while no differences were found in fasting urinary calcium and hydroxyproline excretion. Women without fractures showed a significant correlation between ALP and both age and years since menopause (YSM). Such a correlation is lacking in the fracture group. CONCLUSIONS: Women with vertebral deformities likely due to a fracture had a forearm BMD and markers of bone formation lower than normal. Whether low bone density is due to a low peak of bone mass or to an increased postmenopausal bone loss sustained by an uncoupling between the two bone remodelling processes is still unclear. PMID- 9391997 TI - Influence of weight and gonadal status on total and regional bone mineral content and on weight-bearing and non-weight-bearing bones, measured by dual-energy X-ray absosorptiometry. AB - OBJECTIVE: To evaluate the influence of weight on total body bone mineral content (BMCTB) and regional body bone mineral content (head, arms, trunk and legs). This was studied in accordance with gonadal status and the weight-bearing or non weight-bearing status of each region. METHODS: The study included 94 postmenopausal women (mean age 60.6 +/- 10.5 years), 36 perimenopausal women (mean age 49.0 +/- 2.3 years) and 60 premenopausal women (mean age 36.1 +/- 6.9 years). Full-body bone densitometry (DXA), for measuring total body bone and regional bone mineral content, was carried out in all the women. RESULTS: Among these groups, the influence of 1 kg of body weight on total and regional bone mineral content (percent) did not differ (paired test P ns). In the overall group of women, paired comparison showed differences between the head and other zones measured (P = 0.036-0.004). In the overall group of women, no differences were found in the percent influence of 1 kg body weight on bone mineral content in any study zone (by ANOVA, Fisher's PLSD post hoc test and the Kruskal-Wallis test). In the overall group of women, Fisher's r to z test revealed a non-significant relationship between weight and the bone mineral content of the head (r = 0.49, P ns) but in every other region the relationship between weight and bone mineral content was significant (r = 0.36-0.54, P < 0.0001 in all). CONCLUSIONS: The effect of body weight on BMCTB and regional did not differ significantly with either gonadal status or weight-bearing (legs) and non-weight-bearing bones (arms). PMID- 9391998 TI - Effects of combined low dose of the isoflavone derivative ipriflavone and estrogen replacement on bone mineral density and metabolism in postmenopausal women. AB - OBJECTIVES: To assess the pattern of biochemical markers of bone metabolism and vertebral bone mineral density in early postmenopausal women treated with combined ipriflavone and low dose conjugated estrogens. METHODS: Bone biochemical markers and vertebral bone density were evaluated in a longitudinal, comparative, 2 year study conducted in postmenopausal women treated with sole calcium supplementation (500 mg/day), or with either ipriflavone (IP) at the standard dose (600 mg/day) plus the same calcium dose, low dose conjugated estrogens (CE) (0.3 mg/day) plus calcium, or low dose IP (400 mg/day) plus low dose CE (0.3 mg/day) plus calcium. The results were analyzed by repeated measures analysis of variance, as appropriate. RESULTS: No modifications of both urinary excretion of hydroxyproline and plasma osteocalcin levels were observed in calcium and in CE treated women, while vertebral bone density significantly decreased (P < 0.0001) in both groups. In IP or IP + CE-treated women, plasma osteocalcin did not show any modification, while urinary hydroxyproline showed a significant (P < 0.05) decrease, that paralleled a significant (P < 0.05) increase in vertebral bone density. CONCLUSION: Postmenopausal IP administration, at the standard dose of 600 mg/day, can prevent the increase in bone turnover and the decrease in bone density that follow ovarian failure. The same effect can be obtained with the combined administration of low dose (400 mg/day) IP with low dose (0.3 mg/day) CE. PMID- 9391999 TI - Endometrial effects of three doses of trimegestone, a new orally active progestogen, on the postmenopausal endometrium. AB - OBJECTIVE: To study the effects of oral trimegestone on endometrial histology and vaginal bleeding when given in combination with oral 17-beta-oestradiol. METHODS: This was a prospective, randomised, double-blind, parallel groups, pilot comparative study. Thirty-eight healthy postmenopausal women with normal endometrial histology were given oral 17-beta-oestradiol, 2 mg/day for three continuous cycles of 28 days, plus oral trimegestone, 0.10, 0.25 or 0.50 mg/day from day 15 to day 28 of each cycle. A Vabra biopsy was performed late in the oestradiol/trimegestone phase of cycle 3 and examined for histological evidence of secretory transformation of the endometrium. Characteristics of vaginal bleeding were recorded on a daily basis. RESULTS: Thirty-seven women completed the study, of whom 31 yielded adequate tissue for histological assessment. All showed secretory transformation of the endometrium. Bleeding was of earlier onset and longer duration with the lowest dose of trimegestone. CONCLUSIONS: Trimegestone is a highly effective oral progestogen for endometrial protection, all doses inducing secretory endometrial transformation. Although bleeding patterns suggest a weaker effect of the lowest dose used, the minimum effective dose for endometrial protection has still to be determined and may be lower than those used in this study. PMID- 9392000 TI - Palo Alto Medical Foundation, Research Institute. PMID- 9392001 TI - Recent advances in the treatment of malignant melanoma with gene therapy. PMID- 9392002 TI - Altered procollagen mRNA expression during the progression of avian scleroderma. AB - BACKGROUND: Spontaneous animal models of human autoimmune diseases provide the means to study the very first pathogenetic events, which is not possible in their human counterparts. This is particularly true for connective tissue diseases in which clinical symptoms become manifest only after a long and still obscure course of immunologic, inflammatory, and fibrotic processes. University of California at Davis line 200 chickens (UCD-200) develop a hereditary scleroderma like disease resembling the entire spectrum of human systemic sclerosis, such as early endothelial cell damage, severe lymphocytic infiltration, and accumulation of collagen in skin and internal organs. MATERIALS AND METHODS: In the present study, we investigated mRNA levels of alpha 1(I), alpha 2(I), alpha 1(II), alpha 1(III), alpha 1(VI), alpha 2(VI), and alpha 3(VI) procollagen and GAPDH using digoxigenin-labeled antisense probes in a nonradioactive ribonuclease protection assay (RPA). We analyzed tissue samples from comb, esophagus, heart, lung, and liver of UCD-200 chickens at different stages of the disease, and healthy UCD-058 chickens. RESULTS: During the early inflammatory stage of the disease, the ratios of procollagen types VI/I and types VI/III increased 7-fold in comb tissue, followed by a 3-fold elevation in type I procollagen transcripts in the late acute stage. In the chronic stage, alpha 1(III) procollagen message was increased 2-fold. Additionally, hybridization with the 180 bp alpha 2(I) antisense probe resulted in two bands of 180 bp and 115 bp, respectively, in the RPA. The ratio of these two previously undescribed bands changes in the early stage of the disease both in comb and esophagus. CONCLUSIONS: In an animal model with a spontaneous scleroderma-like disease we found a characteristic, sequential increase in type VI, type I, and type III procollagen transcripts, and we found evidence for the presence and altered ratio of two mRNA variants of alpha 2(I) procollagen, possibly caused by alternative splicing. Comparative analysis of alpha 2(I) procollagen variants in early stages of avian scleroderma and human SSc might provide answers to unresolved questions concerning the molecular basis for generalized fibrosis in scleroderma. PMID- 9392003 TI - Linkage and association studies between the melanocortin receptors 4 and 5 genes and obesity-related phenotypes in the Quebec Family Study. AB - BACKGROUND: The agouti yellow mouse shows adult onset of moderate obesity and diabetes. A depressed basal lipolytic rate in adipocytes or a decreased adrenergic tone arising from antagonizing alpha-melanocyte-stimulating hormone (MSH) activation of melanocortin receptors (MCR) could be at the origin of the obesity phenotype. MATERIAL AND METHODS: MCR 4 and 5 (MC4R, MC5R) genes were studied in the Quebec Family Study. Sequence variations were detected by Southern blot probing of restricted genomic DNA, and mRNA tissue expression was detected by RT-PCR. Subjects with a wide range of weight were used for single-point sib pair linkage studies (maximum of 289 sibships from 124 nuclear families). Analysis of variance across genotypes in unrelated males (n = 143) and females (n = 156) was also undertaken. Body mass index (BMI), sum of six skin-folds (SF6), fat mass (FM), percent body fat (%FAT), respiratory quotient (RQ), resting metabolic rate (RMR), fasting glucose and insulin, and glucose and insulin area during an oral glucose tolerance test were analyzed. RESULTS: MC4R showed polymorphism with NcoI, and MC5R, with PstI and PvuII, with a heterozygosity of 0.38, 0.10, and 0.20, respectively. Linkages were observed between MC5R and BMI (p = 0.001), SF6 (p = 0.005), FM (p = 0.001), and RMR (p = 0.002), whereas associations were observed in females between MC5R and BMI (p = 0.003), and between MC4R and FM (p = 0.002) and %FAT (p = 0.004). After correction for multiple tests, these p values are lowered by one tenth. MC4R and MC5R mRNAs have been detected in brain, adipose tissue, and skeletal muscle. CONCLUSIONS: MC4R and MC5R exhibit evidence of linkage or association with obesity phenotypes, but this evidence is strongest for MC5R. PMID- 9392005 TI - Inducible nitric oxide synthase and proinflammatory cytokine expression by human keratinocytes during acute urticaria. AB - BACKGROUND: IgE/allergen-dependent activation of skin mast cells is involved in acute urticaria and leads to their IL-4 release. Previously we have demonstrated in vitro the induction of the low-affinity receptor for IgE (CD23/Fc epsilon RII) in human keratinocytes (HK) upon stimulation with IL-4. In addition, we have observed that ligation of CD23 on keratinocytes induced type II nitric oxide synthase (iNOS), leading to the release of nitric oxide (NO) and proinflammatory cytokines (TNF-alpha, IL-6). According to these in vitro data, we explored whether keratinocytes could also express iNOS, TNF-alpha, IL-6, and CD23 in acute urticaria, an in vivo model in which activation of mast cells by IgE/allergen immune complexes is involved. MATERIALS AND METHODS: INOS, TNF-alpha, IL-6, and CD23 expression by keratinocytes was studied in acute urticaria (n = 11) in biopsies from lesional and autologous normal skin by immunohistochemistry, in situ hybridization, or RT-PCR. Nitrites and TNF-alpha synthesis were assayed in supernatants of cultured lesional keratinocytes. RESULTS: INOS mRNA expression was demonstrated with RT-PCR in 10 biopsies out of 11 sections of acute urticaria lesional skin. Immunohistochemistry showed that this iNOS positivity originated from keratinocytes located close to the dermoepidermal junction; TNF-alpha and IL 6 mRNA transcription was observed in all but one iNOS+ biopsy. Immunostaining and in situ hybridization with CD23-specific probes were strong in all but one iNOS+ skin biopsy. Noninflamed autologous skin was negative for iNOS (except for a weak positivity in one case), cytokines, and CD23. CONCLUSION: The colocalization of iNOS, proinflammatory cytokines, and CD23 within keratinocytes in acute urticaria demonstrates that these cells play an important role in the initiation and maintenance of the inflammatory reaction during this disease in humans through activation of the iNOS pathway by CD23 ligation with IgE/allergen immune complexes. PMID- 9392004 TI - Constitutive modulation of Raf-1 protein kinase is associated with differential gene expression of several known and unknown genes. AB - BACKGROUND: Raf-1, a cytoplasmic serine/threonine protein kinase, plays an important role in mitogen- and damage-responsive cellular signal transduction pathways. Consistent with this notion is the fact that constitutive modulation of expression and/or activity of Raf-1 protein kinase modifies cell growth, proliferation, and cell survival. Although these effects are controlled at least in part by transcriptional mechanisms, the role of Raf-1 in the regulation of specific gene expression is unclear. MATERIALS AND METHODS: Differential display of mRNA was used to identify the genes differentially expressed in human head and neck squamous carcinoma cells (PCI-06A) transfected with either the antisense c raf-1 cDNA (PCI-06A-Raf(AS)), or a portion of cDNA coding for the kinase domain of Raf-1 (PCI-06A-Raf(K)). The differentially expressed fragments were cloned and sequenced, and they were used as probes to compare the expression patterns in parent transfectants by Northern blot analysis. In addition, expression patterns of the novel genes were examined in normal tissues and cancer cell lines. RESULTS: Six differentially expressed cDNA fragments were identified and sequenced. Northern blot analysis revealed that four of these fragments representing human alpha 1-antichymotrypsin (alpha 1-ACT), mitochondrial cytochrome c oxidase subunit II (COX-II), and two as-yet unidentified cDNAs (KAS 110 and KAS-111) were relatively overexpressed in PCI-06A-Raf(AS) transfectants compared with PCI-06A-Raf(K) transfectants. The other two cDNA fragments representing human elongation factor-1 alpha (HEF-1 alpha) and ornithine decarboxylase antizyme (OAz) were overexpressed in PCI-06A-Raf(K) transfectants compared with PCI-06A-Raf(AS) transfectants. The KAS-110 (114 bp) and KAS-111 (202 bp) cDNAs did not show significant matches with sequences in the GenEMBL, TIGR, and HGS DNA databases, and these may represent novel genes. The KAS-110 and KAS-111 transcripts, approximately 0.9 kb and approximately 0.5 kb, were observed in most normal tissues and several cancer cell types, indicating their housekeeping function. CONCLUSIONS: This study reports novel components of the Raf-1 signaling pathway. alpha 1-ACT, HEF-1 alpha, COX-II, and OAz have been previously implicated in diverse cellular responses including transformation, energy metabolism, and cell survival. Our data suggest that expression of these genes may play a role in the Raf-1-mediated biological activity of PCI-06A cells. The KAS-110 and KAS-111 cDNAs represent unknown genes, and further investigations are necessary to determine their role in the cellular response. Identification of specific targets may provide useful markers for prognosis and therapy selection in squamous cell carcinoma. PMID- 9392007 TI - Report on the first international symposium on oculopharyngeal muscular dystrophy. PMID- 9392009 TI - Hereditary ptosis of late onset: early observations on oculopharyngeal muscular dystrophy in Quebec by Roma Amyot. AB - In 1948, Roma Amyot, a well-known French-Canadian neurologist, observed in ten families a late onset syndrome consisting of hereditary ptosis which was sometimes associated with dysphagia but rarely with limb weakness. At that time, Taylor's original work dating back to 1915 was still unknown to him. Nonetheless, these reports constitute the two earliest publications about this syndrome prevalent in the French-Canadian population. Amyot recognized the myopathic nature of this disease which was later called oculopharyngeal muscular dystrophy (OPMD) by Victor et al. PMID- 9392008 TI - Andre Barbeau and the oculopharyngeal muscular dystrophy in French Canada and North America. AB - Andre Barbeau (1931-1986) is best known world-wide in the neurologic community for his contributions to the study of Parkinson's disease, Huntington's chorea and Friedreich's ataxia. But in Quebec, Canada, his name is associated with oculopharyngeal muscular dystrophy (OPMD), often called here 'maladie de Barbeau', on which he conducted a series of genealogic, genetic and clinical studies early in his career, most intensively from 1964 to 1966. He then demonstrated that most of the reported cases in North America could be traced back to French-Canadian ancestors. Furthermore, he identified this ancestor couple and linked them with a probable case in Niort, in France. Because he was the first to see over a hundred patients, his clinical studies were definitive. He did little work on OPMD after 1967 when he rushed back to the study of L-DOPA in the treatment of Parkinson's disease, a work that he had previously so brilliantly pioneered. PMID- 9392006 TI - Characterization of new polyclonal antibodies specific for 40 and 42 amino acid long amyloid beta peptides: their use to examine the cell biology of presenilins and the immunohistochemistry of sporadic Alzheimer's disease and cerebral amyloid angiopathy cases. AB - BACKGROUND: In Alzheimer's disease (AD), the main histological lesion is a proteinaceous deposit, the senile plaque, which is mainly composed of a peptide called A beta. The aggregation process is thought to occur through enhanced concentration of A beta 40 or increased production of the more readily aggregating 42 amino acid-long A beta 42 species. MATERIALS AND METHODS: Specificity of the antibodies was assessed by dot blot, Western blot, ELISA, and immunoprecipitation procedures on synthetic and endogenous A beta produced by secreted HK293 cells. A beta and p3 production by wild-type and mutated presenilin 1-expressing cells transiently transfected with beta APP751 was monitored after metabolic labeling and immunoprecipitation procedures. Immunohistochemical analysis was performed on brains of sporadic and typical cerebrovascular amyloid angiopathy (CAA) cases. RESULTS: Dot and Western blot analyses indicate that IgG-purified fractions of antisera recognize native and denaturated A beta s. FCA3340 and FCA 3542 display full specificity for A beta 40 and A beta 42, respectively. Antibodies immunoprecipitate their respective synthetic A beta species but also A beta s and their related p3 counterparts endogenously secreted by transfected human kidney 293 cells. This allowed us to show that mutations on presenilin 1 triggered similar increased ratios of A beta 42 and its p 342 counterpart over total A beta and p3. ELISA assays allow detection of about 25-50 pg/ml of A beta s and remain linear up to 750 to 1500 pg/ml without any cross-reactivity. FCA18 and FCA3542 label diffuse and mature plaques of a sporadic AD case whereas FCA3340 only reveals the mature lesions and particularly labels their central dense core. In a CAA case, FCA18 and FCA3340 reveal leptomeningeal and cortical arterioles whereas FCA3542 only faintly labels such structures. CONCLUSIONS: Polyclonal antibodies exclusively recognizing A beta 40 (FCA 3340) or A beta 42 (FCA3542) were obtained. These demonstrated that FAD-linked presenilins similarly affect both p342 and A beta 42, suggesting that these mutations misroute the beta APP to a compartment where gamma-secretase, but not alpha-secretase, cleavages are modified. Overall, these antibodies should prove useful for fundamental and diagnostic approaches, as suggested by their usefulness for biochemical, cell biological, and immunohistochemical techniques. PMID- 9392010 TI - Oculopharyngeal muscular dystrophy, other ocular myopathies, and progressive external ophthalmoplegia. AB - Progressive external ophthalmoplegia comprises many different disorders. Those of childhood onset can be separated from juvenile or adult onset. Among those of later onset the most common causes are oculopharyngeal muscular dystrophy, oculopharyngodistal muscular dystrophy and the several mitochondrial disorders, especially those with large deletions of mitochondrial DNA (mtDNA) (sporadic), those with maternal inheritance (point mutations), or the autosomal dominant forms with multiple deletions of mtDNA. Ophthalmoplegia of presumably neurogenic origin is seen in some of the familial spinocerebellar ataxias. Advances in molecular genetics should provide information about affected gene products and, therefore, pathogenesis. PMID- 9392012 TI - Oculopharyngeal muscular dystrophy in France. AB - The clinical, histopathological, ultrastructural and geographical data on 29 cases of oculopharyngeal muscular dystrophy (OPMD) identified by the authors in France is briefly presented. The mean age of the patients was 53.8 +/- 8.1 years. Onset symptoms were ptosis (14/29), dysphagia (12/29) and limb girdle weakness (3/29). The evolution of the disease was always progressive and followed different clinical patterns. The main histological changes in muscle biopsies were atrophic angulated fibers (29/29) and rimmed vacuoles (25/29); muscle fiber necrosis was very rare (1/29). The characteristic nuclear inclusions made of 8.5 nm filaments were observed in all cases, and found in 2-5% of the nuclei in a given ultrathin section. They are the morphological marker of the disease. PMID- 9392011 TI - Recent studies on oculopharyngeal muscular dystrophy in Quebec. AB - In 1990, we launched a major study to ascertain the clinical picture of OPMD in Quebec and to identify large families for linkage analysis. In 14 patients, the chromosomes were karyotyped to eliminate any deletion or translocation. Relevant family information and clinical data were computerized and correlations were sought for the age of onset, the identification of the first symptom and the distribution of weakness. A simple test to detect dysphagia was validated. Twenty one families have taken part in the study, which led to our localization of the gene in 1995 [Brais B, Xie Y-G, Sanson M, et al. Hum Mol Genet 1995; 4:429-434]. At least one case in each family underwent muscle biopsy to confirm the presence of the typical nuclear filaments found in OPMD. Electrodiagnostic and pathologic studies were also conducted to better understand the disease process. An illustrative case is presented. PMID- 9392013 TI - Genealogical study of oculopharyngeal muscular dystrophy in France. AB - This work is based on 54 probands affected by oculopharyngeal muscular dystrophy (OPMD). The muscle biopsy of all these patients showed the presence of the intranuclear inclusions, specific of this disease. The residence of the probands is concentrated in three clusters: the Paris, Marseilles and Bordeaux regions. The genealogical study was carried out on 43 probands, 10 of which did not have any ascendance in France for more than two generations. The geographic origin of the 33 patients of French descent was distributed over numerous regions, not including the Paris and Marseilles regions where many patients lived. This geographic dispersion and the rarity of common genealogies of the probands, did not suggest the existence of a recent founder effect, in contrast to what is observed in the French-Canadian community. The existence of a link between French and French-Canadian families is currently being investigated. PMID- 9392014 TI - Epidemiology and inheritance of oculopharyngeal muscular dystrophy in Israel. AB - Oculopharyngeal muscular dystrophy (OPMD) is considered frequent among French Canadians. Our previous observations suggested it is common also among the Jews originating from Bukhara in Uzbekistan, many of whom are now living in Israel. One hundred and seventeen OPMD patients were identified in a population of 70,000 people of Bukharian descent, resulting in a calculated minimal prevalence of 1:600. In all but three families age dependent autosomal dominant inheritance was documented. There is some evidence for genetic anticipation. Three young, severely ill, patients from two different families may be homozygotes, their parents being both affected. Bukhara Jews present the second largest known cluster and the prevalence is the highest in the world. The existence of very large families, intermarriage among carriers and probably homozygote offspring may be useful for genetic studies. A 'founder effect' may explain the high prevalence of OPMD in this population. PMID- 9392015 TI - Oculopharyngeal muscular dystrophy in Japan. AB - Oculopharyngeal muscular dystrophy (OPMD) in the European population has been frequently diagnosed, but except for one black family, the occurrence in other ethnic groups is uncertain. We identified two unrelated OPMD Japanese families, including 34 affected individuals. Major clinical manifestations were bilateral ptosis and dysphagia starting after age 40. Histologic studies of limb muscles revealed mild myogenic changes, occasional rimmed vacuoles, and small angulated fibers. By contrast, cricopharyngeal muscle showed a marked loss of fibers and massive proliferation of connective tissue. Intranuclear tubulofilamentous inclusions (ITFI) of 8.5 nm outer diameter were observed in 2-5% of the nuclei in four different biopsied muscles. One patient with recurrent aspirations underwent successful cricopharyngeal myotomy. Aerodynamic examination was useful to evaluate velopharyngeal closure function. Our investigations revealed that OPMD is a geographically widespread disorder, and ITFI may be the specific morphologic hallmark. PMID- 9392016 TI - Oculopharyngeal muscular dystrophy in Uruguay. AB - Within the last 30 years, sixty-five patients exhibiting the clinical symptoms of oculopharyngeal muscular dystrophy (OPMD) were studied at the Neuromuscular Diseases Unit of the Neurological Institute of Montevideo. They are members of five unrelated families which came from the Canary Islands to Uruguay between 1850 and 1900. In the three families examined, the typical inclusions characteristic of OPMD were found in the nuclei of muscle fibers. Treatment for ptosis and dysphagia was discussed. The particular migratory pattern of this group of patients could be of considerable interest in the study of molecular genetics. PMID- 9392017 TI - Oculopharyngeal muscular dystrophy in Italy. AB - Oculopharyngeal muscular dystrophy (OPMD) is an autosomal dominant myopathy particularly frequent in Quebec. The few Italian cases thus far described with bilateral ptosis, dysphagia and variable muscle weakness, show non-specific dystrophic findings on muscle biopsies by light microscopy. We describe a 70-year old Italian woman with an adult-onset ptosis, mild dysphagia and proximal muscle weakness belonging to a family segregating OPMD according to an autosomal dominant mode of inheritance. Clinical features of four of her relatives are reviewed. Muscle biopsy studied by electron microscopy showed the typical 8.5 nm in diameter intranuclear filamentous inclusions (INI). To our knowledge, this is the first Italian report of OPMD with INI. The identification of nuclear inclusions is mandatory in order to confirm the diagnosis prior to linkage analysis. PMID- 9392018 TI - Oculopharyngeal muscular dystrophy in a northern German family linked to chromosome 14q, and presenting carnitine deficiency. AB - We report the evaluation of oculopharyngeal muscular dystrophy (OPMD) in a large northern German family, which can be traced back six generations and is unrelated to French-Canadian families. The symptoms in this family start at about 50 years of age and include dysphagia, bilateral ptosis, and in some cases a slowly progressive atrophy and weakness of other extraocular, facial or limb girdle muscles. The muscle biopsies showed the pathognomonic ultrastructural finding of characteristic intranuclear filaments. Linkage analysis confirmed that this family is also linked to chromosome 14q markers. Haplotype analysis revealed that a unique haplotype segregates with the disease which is different from the one found in French-Canadian OPMD. Although approximately half of the probands with OPMD showed mild clinical and neurophysiological signs of a distal symmetrical neuropathy, the association between the neurogenic lesions and OPMD is still unclear. Some family members with or without OPMD complained of exercise related muscle pain, and a lipid storage myopathy with low muscular carnitine concentrations was found, while the carnitine contents in blood and urine samples as well as the activity of the carnitine-palmitoyl-transferase were normal, fitting the pattern of a primary muscular carnitine deficiency, independent of OPMD. PMID- 9392019 TI - Morphological changes in muscle fibers in oculopharyngeal muscular dystrophy. AB - The study of muscle biopsies of 29 cases of oculopharyngeal muscular dystrophy (OPMD) showed the two main morphological features of this disease: rimmed vacuoles (in 26 cases) and intranuclear inclusions (in all cases). These inclusions are made of 8.5 nm tubular filaments and the areas occupied by them are lighter than the surrounded nucleoplasm. This can be seen by light microscopy, facilitating the detection of the tubulo-filamentous inclusions which can only be identified with certitude by electron microscopy. In a given ultrathin section the area occupied by these inclusions varied from 2% to 5% of the nuclei. The intranuclear inclusions are the morphological marker of OPMD and their finding in a muscle biopsy allows the exact diagnosis of this disease. The origin and biochemical nature of the intranuclear inclusions is unknown. PMID- 9392020 TI - Using the full power of linkage analysis in 11 French Canadian families to fine map the oculopharyngeal muscular dystrophy gene. AB - Oculopharyngeal muscular dystrophy (OPMD) is a late onset autosomal dominant muscular dystrophy with a high prevalence in the French Canadian population. We report linkage analysis with 7 chromosome 14q polymorphic markers in 11 large French Canadian families. An observed recombination in one family establishes D14S283 as the new centromeric flanking marker, therefore reducing the previously reported candidate interval from 5cM to 2cM. The highest two-point LOD score was 26.05 at theta = 0.01 for MYH7.1. Multipoint analysis suggested that the OPMD genes lies within a 1.5cM region around D14S990. This study of large French Canadian families underlines the great power of this population to fine map disease genes. PMID- 9392021 TI - Confirmation of linkage of oculopharyngeal muscular dystrophy to chromosome 14q11.2-q13 in American families suggests the existence of a second causal mutation. AB - Oculopharyngeal muscular dystrophy (OPMD) is a late-onset, autosomal dominant disorder characterized by progressive ptosis, dysphagia, and extremity weakness. Linkage of OPMD to 14q11.2-q13 has been reported in a series of French-Canadian families. Tightly linked markers have been defined and haplotype analysis in these data show a single segregating disease chromosome throughout the OPMD French-Canadian families. We have ascertained and sampled five multigenerational outbred American OPMD families. Four of the five families have known French Canadian ancestry while the fifth is of English/Scottish origin. Linkage analysis was performed using standard likelihood methods. A peak multipoint lod score of 6.30 was obtained for the marker MYH7.1 in the OPMD families. The English/ Scottish family exhibited a different chromosomal haplotype for the OPMD alleles than the families of French-Canadian origin. These data suggest this family may represent a second, possibly independent mutation in this disorder. Linkage was confirmed to chromosome 14q11.2-q13 with no evidence of genetic heterogeneity. PMID- 9392022 TI - Surgical correction of blepharoptosis in oculopharyngeal muscular dystrophy. AB - Progressive, usually symmetric blepharoptosis with or without dysphagia appears in most instances in the fifth decade in oculopharyngeal muscular dystrophy (OPMD). We review our experience over 20 years of applying Beard's surgical guidelines for correction of ptosis to OPMD patients with satisfactory results. As the disease continues to progress, the rate of recurrence of ptosis among follow-up patients of a 9-year minimum period was 13%. PMID- 9392023 TI - Cricopharyngeal myotomy in the management of neurogenic and muscular dysphagia. AB - Oropharyngeal dysphagia results from disruption of the integrated mechanism of swallowing. Neurogenic dysphagia is caused by central nervous system disorders or by cranial nerve involvement and it may be distinguished from muscular dysphagia such as that seen mostly in oculopharyngeal muscular dystrophy (OPMD). Based on our 20-year experience in a university hospital thoracic surgery service, we describe the results of the clinical evaluation, the laboratory testing and the surgical management of a recent subgroup of patients experiencing dysphagia from neurogenic and muscular disorders. PMID- 9392024 TI - Upper esophageal sphincter myotomy in oculopharyngeal muscular dystrophy: long term clinical results. AB - From 1980 to 1995, 53 patients with oculopharyngeal muscular dystrophy (OPMD) underwent an upper esophageal sphincter (UES) myotomy for the control of marked dysphagia. From this number, a group of 21 patients had been evaluated for preoperative and postoperative symptoms in 1987. The same clinical assessment was performed in 1995 by an independent evaluator for a total of 37 patients including 12 patients from the first group. As a whole, after a mean follow-up of 6.2 years, surgery succeeded in 18 patients (49%), gave a partial improvement in 12 (32%) and failed in seven (19%). The 12 patients evaluated twice (in 1987 and 1995) have had very good early results, 8-69 months after UES myotomy: dysphagia was totally relieved in eight patients, occurred rarely in three and was moderate in one. Nevertheless, the very long-term follow-up (8 years later) has shown a recurrence of the swallowing and tracheobronchial symptoms in many cases. PMID- 9392025 TI - Dysphagia in oculopharyngeal muscular dystrophy: a series of 22 French cases. AB - Twenty-two patients (mean age = 67.9 years) with oculopharyngeal muscular dystrophy (OPMD) were referred for dysphagia from 1987 to January 1995. Six patients had suffered aspiration pneumonia, and three had significantly lost weight, while 19 complained of discomfort during swallowing but without weight loss. Swallowing was assessed by fiberscopy during swallowing (last eight patients), videofluoroscopy (12 cases) and manometry (19 cases). Twelve patients underwent a cricopharyngeal (CP) myotomy: 10 showed improvement, one had a partial improvement, and the procedure failed in one (mean follow-up = 29.6 months). In the other cases, CP myotomy was postponed, refused or contraindicated. Of the 22 patients, three died from OPMD consequences. Factors associated with favorable outcome were adequate residual pharyngeal propulsion and no weight loss. In a majority of cases, CP myotomy constitutes an effective treatment of dysphagia with adequate residual propulsion but does not modify the final prognosis and is contraindicated in cases with pharyngeal aperistalsis. PMID- 9392027 TI - Attempting to fathom the unfathomable: descriptive views of spirituality. AB - OBJECTIVES: To examine the various views of "spirituality and its uses in the disciplines of philosophy, theology, psychology, and nursing. DATA SOURCES: Definitions and descriptions of spirituality and related terms from the disciplines of philosophy, theology, psychology, and nursing. CONCLUSION: Although it is widely accepted that holistic nursing care incorporates care of the spirit, nursing's view of spirituality is influenced by varying paradigms. Nursing researchers are exploring spirituality. Spiritual care that is ethical and sensitive is an invaluable part of total patient care. IMPLICATIONS FOR NURSING PRACTICE: Appreciating various views of spirituality and recognizing the possible discrepancy between a nurse's and a patient's view of spirituality allows the reader to use such terms carefully and appropriately in providing sensitive patient care. PMID- 9392026 TI - A pilot study on upper esophageal sphincter dilatation for the treatment of dysphagia in patients with oculopharyngeal muscular dystrophy. AB - Upper esophageal sphincter (UES) dilatation was done for the treatment of moderate to severe dysphagia with a Maloney bougie in 14 patients with oculopharyngeal muscular dystrophy (OPMD) or with an achalasia dilator in three patients. The severity of dysphagia prior to UES dilatation was evaluated by a 15 point dysphagia score, a pharyngeal and esophageal manometry and a radionuclide pharyngoesophageal transit study. Using actuarial life table, the improvement rate after dilatation with Maloney bougie was 64.3% (95% CI 39.2-89.4) at 3- and 6-month follow-ups, and was 55.7% (95% CI 28.9-82.5) at 12- and 18-month follow ups. At 3-month post-dilatation, we observed a significant reduction of the mean dysphagia score from 9.6 to 7.2 (P = 0.05). No significant manometric or radionuclide factors were found to predict effective dilatation. The results of this pilot study showed that UES dilatation with Maloney bougie or achalasia dilator may be an effective treatment of moderate dysphagia in patients with OPMD. However, further studies with larger sample sizes are needed to corroborate these results and to assess long-term outcome. PMID- 9392028 TI - Cultural aspects of spirituality in cancer care. AB - OBJECTIVES: To recognize the significant relationship between culture and spirituality; to offer techniques for assessing the influence of culture on spirituality. DATA SOURCES: Books and journal articles from nursing and anthropology. CONCLUSION: Patients' spirituality may be determined entirely by cultural norms, in opposition to cultural norms, or by both these norms and individual life experiences. A thorough assessment of a patient's spirituality with careful attention to the degree to which that spirituality is affected by the patient's cultural background is essential. IMPLICATIONS FOR NURSING PRACTICE: Nurses who care for persons with cancer need to undertake a process of self-assessment of their spirituality before providing spiritual care for patients. Such an assessment helps to prevent cultural imposition. PMID- 9392030 TI - The arts in spiritual care. AB - OBJECTIVES: To explore the role of the arts in spirituality and spiritual care and the importance of the arts and creativity in health care settings, particularly where individuals are confronting life-threatening illnesses. DATA SOURCES: Professional and lay journals/magazines, and personal experience with oncology and hospice patients. CONCLUSION: The arts are now viewed as an integral component of holistic care for patients and families. By offering opportunities to engage in the arts and creative expression, persons with cancer can be enabled to mourn, grieve, celebrate life, be empowered to endure their situation, and find healing and meaning. IMPLICATIONS FOR NURSING PRACTICE: Comprehensive supportive care for cancer patients requires the efforts of an interdisciplinary team. Artists can play a role as a part of this team. Oncology nurses must be knowledgeable of the role of the arts and creative expression in the provision of care to patients with cancer and how to incorporate the arts into the cancer care setting. PMID- 9392029 TI - Spiritual assessment across the cancer trajectory: methods and reflections. AB - OBJECTIVES: To provide information about spiritual assessment strategies and the spiritual stages through which persons with cancer pass. DATA SOURCES: Books and articles (including research reports) from various disciplines including nursing, medicine, theology, and other health care professions: personal narratives and reflections of individuals with cancer. CONCLUSION: Sound spiritual assessment is prerequisite for sound spiritual intervention. Nurses must find the time, means, and knowledge to incorporate spiritual assessment into nursing care. Patients agree. IMPLICATIONS FOR NURSING PRACTICE: Nurses can improve spiritual assessment by eliciting patient accounts of the evolving spiritual journey and prayers that parallel changes in health status. PMID- 9392031 TI - Constructing meaning from the experience of cancer. AB - OBJECTIVES: To describe how people with cancer create new sources of meaning at significant times in the illness trajectory. DATA SOURCES: Nursing research, including that of the author; documented observations and theories of psychiatrists and psychologists. CONCLUSION: There are turning points within the cancer trajectory when choices made by persons with cancer and their families may have far reaching consequences in terms of new goals to pursue and new sources of meaning to create. IMPLICATIONS FOR NURSING PRACTICE: Oncology nurses can assist by carefully listening to what is important to patients, assisting to clarify values when necessary, and encouraging patients to seek connections with similar others that facilitate finding meaning and healing. PMID- 9392032 TI - The story behind the story: the use of storytelling in spiritual caregiving. AB - OBJECTIVES: To briefly discuss the nature and function of stories that patients tell, and offer practical tips on how to listen and make sense of these stories. DATA SOURCES: Books and articles from disciplines in the humanities and health care professions. CONCLUSION: Stories are a medium for assessment and intervention in areas that essentially reflect an individual's spirituality. IMPLICATIONS FOR NURSING PRACTICE: Encouraging storytelling is an intervention nurses can use to promote spiritual health. Suggestions for eliciting and analyzing stories are offered. PMID- 9392033 TI - Replenishing the spirit by meditative prayer and guided imagery. AB - OBJECTIVES: To review relevant literature describing prayer and guided imagery, and to demonstrate via the use of a vignette, the use of both prayer and guided imagery as one approach to offer spiritual care to oncology patients. DATA SOURCES: Review and research articles from multiple disciplines, and personal clinical experience. CONCLUSION: Meditative prayer and guided imagery are two approaches that can be used to provide spiritual care to cancer patients and families. While research has focused on elements of spirituality, research related to clinical interventions is limited. IMPLICATIONS FOR NURSING PRACTICE: Guided imagery, metaphors, meditative prayer, and prayers of silence are effective approaches the nurse can implement when caring for the patient with cancer. PMID- 9392034 TI - The practice of presencing. AB - OBJECTIVES: To understand philosophical foundations for this fundamental nursing intervention, and to review literature and expert nurses' experiences to describe how to be present to persons with cancer. DATA SOURCES: Author's qualitative research of nurses' experiences of presencing; theoretical and clinical nursing perspectives; books by theologian Martin Buber. CONCLUSION: Presencing requires deliberate focused attention, receptivity to the other person, and persistent awareness of the other's shared humanity. "Being there" provides comfort for both patient and nurse. IMPLICATIONS FOR NURSING PRACTICE: Literature reviewed offer the elements of presenting (eg, providing affirmation, communication of empathy, being without words). Identification of such elements can assist the reader to implement this intervention. PMID- 9392035 TI - Spiritual care for children with cancer. AB - OBJECTIVES: To review literature pertinent to spirituality of children with cancer and to identify practical strategies for providing care for this dimension in children. DATA SOURCES: Nursing research and literature about pediatric nursing care and spirituality; theoretical formulations of Piaget, Fowler, and Erikson. CONCLUSION: Children diagnosed with cancer have unique spiritual needs that place them at risk for developing spiritual distress. With the diagnosis may come experiences of loss of normalcy, physical stamina, relationships, body image, and future goals. Spiritual care includes interventions that assist children to find meaning and purpose in life, to continue relationships, and to transcend beyond the self. IMPLICATIONS FOR NURSING PRACTICE: Spiritual care includes caregiver and child assessment and interventions appropriate to the developmental stages of infancy through adolescents. Tables outlining how this can be done by oncology nurses are included. PMID- 9392036 TI - Spiritual care: the needs of the caregiver. AB - OBJECTIVES: To explore the spiritual needs of the family caregiver and to provide suggestions for giving spiritual care to this caregiver. DATA SOURCES: A caregiver's personal experience and nursing texts. CONCLUSION: Providing care for a loved one with cancer can be stressful for the family caregiver; yet, it can also produce spiritual growth. By providing care for the caregiver, the oncology nurse is equipping this caregiver to address the spiritual needs of the patient. IMPLICATIONS FOR NURSING PRACTICE: Nurses can assist caregivers by offering actions that communicate love, support, acceptance, and faithfulness. Such measures can ease the pain and encourage spiritual wellness. PMID- 9392037 TI - Healing partners: the oncology nurse and the parish nurse. AB - OBJECTIVES: To explore the role of the parish nurse and to examine elements of a parish nurse program and standards of care. DATA SOURCES: Journal articles, book chapters, and personal experience related to parish nursing. CONCLUSION: Parish nursing is a fairly new specialized area of nursing with standards of care and professional performance. The parish nurse concept has gained acceptance across the country and the number of parish nurse programs are increasing. IMPLICATIONS FOR NURSING PRACTICE: The parish nurse focuses on health promotion within the context of the beliefs, values, and practices of a faith community. The role of the parish nurse centers on providing education and support to members of a congregation. Oncology nurses and parish nurses can work together as partners in providing care to patients with cancer and their families. PMID- 9392038 TI - Collaboration between nurses and chaplains for spiritual caregiving. AB - OBJECTIVES: To discuss three ambiguities that may accompany nurse-chaplain collaboration in providing spiritual care: confusion about the meaning of "spiritual" and related terms, spiritual assessment and the referral process, and the role of clergy. DATA SOURCES: Review and research articles related to nursing and pastoral care, and documented standards. CONCLUSION: Effective nurse-chaplain collaboration is necessary (especially considering current health care system changes) to provide adequate spiritual care. Additionally, the increased involvement of both nurses and chaplains in ethical issues is likely to make nurse-chaplain collaboration increasingly important. IMPLICATIONS FOR NURSING PRACTICE: Nurses must collaborate with chaplains and relate to clergy to provide spiritual care for cancer patients and families. Knowledge of assessment differences between nurses and chaplains, terminology, and role of clergy will enhance this collaboration. PMID- 9392039 TI - A report on the development and work of the Labor Welfare Corporation Spinal Injuries Center in Japan. PMID- 9392040 TI - Rehabilitation of spinal cord injury in the national rehabilitation center for the disabled of Japan: profile of a spinal service. AB - The National Rehabilitation Center for the Disabled (hereunder abbreviated NRC) in Japan was established in 1979. It consists of four divisions: the hospital, the rehabilitation training center, the research institute, and the information service section. The spinal unit has been functioning and cooperating corelatively with all of these divisions. There were 1047 patients with a spinal cord injury treated in the 15 years from September, 1980 to August, 1994; consisting of 924 males (88.3%), and 123 females (11.7%). The breakdown of causes of injury were traffic accidents 44.9%, having a fall 14.7%, sports accidents 6.7%, and other causes of spinal paralysis 10.5%. The sites of the spinal cord lesions were cervical 372 (35.5%), thoracic 547 (52.5%), and lumbar spinal cord 128 patients (12.3%). The incidence of complete paralysis in those with a cervical spinal cord injury (SCI) was 68.8%, and for thoracic and lumbar SCI 79% each. The time for completion of activities of daily living (ADL) was 12.0 +/- 1.54 months in those with tetraplegia, and 5.6 +/- 1.71 months for those with paraplegia. The rate of employment for reentry into society was 59% in those with a cervical spinal cord injury, and 74% each in those with a thoracic or lumbar spinal cord injury. PMID- 9392041 TI - Extraforaminal lumbar disc herniation at two contiguous intervertebral levels. AB - We describe two unusual surgical cases who presented with extraforaminal lumbar disc herniation that occurred at two adjacent vertebral levels simultaneously and unilaterally. Magnetic resonance imaging and selective nerve root infiltration followed by radiculography helped to outline the herniated disc material. Lateral fenestration and microsurgical foraminal widening of the affected vertebral levels allowed a complete and safe relief of the compressed nerves. PMID- 9392042 TI - Decreased choline acetyltransferase activity in the murine spinal cord motoneurons under chronic mechanical compression. AB - The tiptoe-walking Yoshimura (twy) mouse is a model of chronic spinal cord compression caused by ossification of intraspinal ligaments. Choline acetyltransferase (CAT), which is known to be a specific marker of cholinergic neurons, best reflects spinal motoneuron function. Changes in CAT immunoreactivity following chronic spinal cord compression in twy mice were investigated quantitatively in order to elucidate spinal motoneuron functional changes according to the degree and direction of compression. Thirty 24-week-old twy mice were used in this study. They were divided into three groups according to the direction of spinal cord compression (anterior, posterior, and lateral) and the CAT immunoreactivities in whole sections of their upper cervical spinal cords were investigated quantitatively using a fluorescence microphotometry system. The lateral compression group showed histological spinal motoneuron atrophy and loss on the compressed, but not the non-compressed, side. Spinal motoneuron atrophy and loss were observed when the severity of spinal canal stenosis due to the ossified lesion, expressed as the occupation rate, was 30% or more, but the spinal motoneurons appeared normal when it was below 30%. The CAT immunofluorescence intensity of the anterior horn showed a linear negative correlation with the degree of canal stenosis. When the occupation rate was below 20%, the CAT immunofluorescence intensities in the anterior horns of the compression and control groups did not differ significantly. The CAT immunofluorescence intensity of twy mice with occupation rates of 20% or more were significantly lower than that of those with occupation rates below 20%. Furthermore, the CAT immunofluorescence intensity was significantly lower on the compressed than the non-compressed side of the lateral compression group. Thus, our findings indicate that an occupation rate of about 20% may be the critical level for functional changes in the spinal motoneurons. PMID- 9392043 TI - Lumbar cerebrospinal fluid pulse wave rising from pulsations of both the spinal cord and the brain in humans. AB - There are two theories regarding the origin of the lumbar cerebrospinal fluid pulse wave (L-CSFPW): that it arises from the arteries supplying the spinal cord, and that it is due to the pulsations of the brain transmitted through the subarachnoid space of the spine. We investigated L-CSFPW of 11 myelopathic patients with a complete (five patients, CB-group) or an incomplete spinal block (six, ICB-group) on myelography to determine the origin of L-CSFPW. Since arterial pressure amplitude (APA), the energy source of L-CSFPW, is not the same between individuals or between before and after operation, not only L-CSFPW itself but also the transfer function between the arterial pressure wave and the L-CSFPW calculated by the system analysis method was analyzed to eliminate the influence of hemodynamic fluctuations. In the system analysis, the arterial pressure wave, L-CSFPW and transfer function were decomposed into five harmonic waves (HW). In the CB group, L-CSFPW was observed to be 0.72 mmHg on average (range, 0.25-1.00) in spite of blocking pulsations of the brain, showing that there was a contribution to L-CSFPW unrelated to the brain, that is, the spinal cord. In the CB group, however, the preoperative transfer function value of HW1 (mean, 0.056; range, 0.012-0.170) was lower than that in the ICB group (mean, 0.137; range, 0.061-0.236) (P < 0.05), indicating that the brain pulsation also contributed to L-CSFPW. In the ICB group, there was significant reduction of HW1 (P < 0.01) and HW2 (P < 0.05) transfer function after posterior decompression surgery in spite of improvement in the subarachnoid space narrowing: preoperative HW1, mean, 0.137, range, 0.061-0.236; postoperative HW1, mean, 0.065, range, 0.021-0.153; preoperative HW2, mean, 0.092, range, 0.011-0.148; postoperative HW2, mean, 0.044, range, 0.030-0.066. It has been reported that the spinal cord blood flow is decreased 20% or more by laminectomy, therefore, L-CSFPW measurement may be sensitive enough to detect a 20% or higher decrease in this flow. This suggests that L-CSFPW could possibly be used clinically as a non invasive method of monitoring the spinal cord blood flow. For broad clinical application of CSFPW, however, further studies are needed, especially on the direct relationship between CSFPW and spinal cord blood flow itself. PMID- 9392044 TI - Experimental chronic compression on the spinal cord of the rabbit by ectopic bone formation in the ligamentum flavum with bone morphogenetic protein. AB - This study was conducted to induce chronic spinal cord compression myelopathy in rabbits. The L5 lumbar lamina was cut partially in 70 rabbits, and bone morphogenetic protein (BMP) was implanted on the ligamentum flavum in 35 of them. In the BMP group, new bone formed on the dorsal side of the spinal canal and flattened the spinal cord in an anteroposterior direction. No pathological changes were detected in the intramedullary tissues by light microscopic examination. In rabbits it is possible to induce compression of the cord by using BMP, although sufficient cord compression to induce myelopathy was not achieved. PMID- 9392045 TI - Later health-related quality of life in adults who have sustained spinal cord injury in childhood. AB - The outcome in terms of health-related quality of life (HRQL) in pediatric spinal cord injury (SCI) was studied in 36 adults who had sustained an SCI in childhood. The patients were interviewed and clinically examined. HRQL was assessed with the 15D, a generic fifteen-dimensional self-administered HRQL instrument. The 15 multiple-level dimensions are moving, seeing, hearing, breathing, sleeping, eating, communicating, urinary continence, working, social participation, mental functioning, pain, depression, distress and perceived health. The respondents choose, for each dimension, the level that best describe their health status. In the 15D valuation system the respondents first assign a relative importance weight to each dimension and then a relative value to the levels on each dimension. To derive the 15D HRQL score on a 0-1 scale the level values and importance weights are multiplied and combined with the levels chosen. The average HRQL score of this SCI group was significantly lower than that measured in the population sample. The average importance weights assigned by the SCI group differed significantly (P < 0.05) from those assigned by the general population on several dimensions. The weights assigned by the SCI group were higher for the dimensions of mental functioning, communicating, social participation and seeing and lower for moving, working, sleeping and eating. These differences in valuing the dimensions of HRQL can influence behaviour and should therefore be taken into consideration in rehabilitation. PMID- 9392046 TI - Muscle reorganization following incomplete cervical spinal cord injury in rats. AB - This study on rats was designed to evaluate the change of biceps muscle fibers by enzyme histochemical examination and the change of distribution of motoneurons innervating biceps muscle fibers by retrograde tracer examination after incomplete spinal cord injury. Incomplete spinal cord injury was produced by placing a 20 g weight on exposed dura at the C6 level. The number of the labeled motoneurons of C6 segment significantly decreased compared with that in the control, but there were no significant changes in the other segments at 4 weeks after the injury. Moreover, there was type grouping of biceps muscle fibers at 4 weeks after the injury. These findings indicated that the incomplete cervical spinal cord injury at C6 level in rats caused the partial denervation and then reinnervation of biceps muscle fibers by the collateral sprouting of the remaining motoneurons which belonged to the same motoneuron pool of the injured motoneurons. PMID- 9392047 TI - Objective evaluation of pain in various spinal diseases: neuropeptide immunoreactivity in the cerebrospinal fluid. AB - A quantitative analysis was performed of substance P-like immunoreactivity (SPLI) and of beta-endorphin-like immunoreactivity (beta-ENDLI), in the cerebrospinal fluid (CSF) in various diseases. The results reported to date have not been consistent. The purpose of this study was to investigate whether or not the concentration of SPLI or that of beta-ENDLI in CSF demonstrated any potential for assessing the degree of subjective pain in various spinal diseases. SPLI in CSF was measured by radioimmunoassay in 158 patients with a spinal disease; involving 57 patients with a lumbar disc herniation (LDH), 38 with lumbar canal stenosis (LCS), 46 with cervical myelopathy (CM) and 17 with cervical radiculopathy (CR), and also in 20 healthy controls. beta-ENDLI in CSF was measured in 25 of these same patients; involving 12 with LDH, seven with LCS and six with CM, and also five of the same controls. The concentration of serum SPLI was also measured in 50 of these 158. The severity of pain was self-evaluated by each patient using a linear visual analogue scale (VAS). Their Japanese Orthopaedic Association (JOA) score was also calculated objectively using the clinical findings. Correlations were investigated among the concentrations of SPLI and beta-ENDLI in the CSF and the VAS and JOA clinical assessments of these patients. The concentration of SPLI in CSF was significantly higher in various spinal diseases than in control (P < 0.05), and was correlated with the severity on the VAS and with the JOA score. However, beta-ENDLI was not correlated with either the VAS or the JOA score. We conclude that the measurement of the SPLI concentration in CSF has the potential for assessing objectively the severity of pain associated with various spinal diseases. PMID- 9392048 TI - Prediction of the surgical outcome for the treatment of cervical myelopathy by using hyperbaric oxygen therapy. AB - The effectiveness of hyperbaric oxygen therapy (HBO) in predicting the recovery after surgery in patients with cervical compression myelopathy was evaluated. HBO has been used to treat brain and spinal cord diseases, but the effect is generally temporary. This is the first paper to utilize HBO as a diagnostic tool to evaluate the functional integrity of the spinal cord. The study group consisted of 41 cervical myelopathy patients aged 32-78 years. Before surgery, the effect of HBO was evaluated and was categorized as four grades. The severity of the myelopathy and the recovery after surgery were evaluated by the score proposed by the Japanese Orthopaedic Association (JOA score). The correlation between many clinical parameters including the HBO effect and the recovery rate of JOA score was evaluated. The recovery rate of JOA score was found to be 75.2 +/- 20.8% in the excellent group, 78.1 +/- 17.0% in the good group, 66.7 +/- 21.9% in the fair group and 31.7 +/- 16.4% in the poor group. There was a statistically significant correlation between the HBO effect and the recovery rate of the JOA score after surgery (r = 0.641, P < 0.0001). The effect of HBO showed a high correlation with the recovery rate after surgery as compared to the other investigated parameters. HBO can be employed to assess the chance of recovery of spinal cord function after surgical decompression. PMID- 9392049 TI - Cervical spondylosis in paraplegic patients and analysis of the wheelchair driving action: a preliminary communication. AB - We investigated cervical spondylotic changes in paraplegic patients by examining the radiographs of their cervical spine. The incidence of cervical spondylosis in these patients was significantly higher than was found in normal control subjects of a matching age. We attributed this finding to the mechanical stress on the cervical spine during wheelchair driving. We then performed a biomechanical analysis using electromyography and goniometer. Paraplegic patients frequently flexed and extended their cervical spine shown by the goniometer, whilst the normal control subjects walked almost without moving their cervical spine. However, the integrated electron myography (IEMG) of the paraplegic patients was not synchronously increased with the integration of goniometer (IGOM) and the IEMG of the normal controls was significantly larger than that found in paraplegic patients on increasing their speed. This was thought to be necessary for steadying the head whilst walking or running. PMID- 9392050 TI - Huge solitary osteochondroma at T11 level causing myelopathy: case report. AB - A solitary osteochondroma of the vertebral column is rare, and also it will rarely cause neurological deficits. Myelopathy from a tumour usually presents insidiously with neurological deficits. We report a case of a huge solitary osteochondroma at T11 level with an acute onset of myelopathy induced by a minor trauma. MRI findings of a spinal osteochondroma has rarely been described. In our patient, the MRI demonstrated an outer osteochondral layer and an ossified centre of the mass. A literature review has also been undertaken. PMID- 9392051 TI - Myelopathy secondary to ossification of the posterior longitudinal ligament of the thoracic spine treated by anterior decompression and bony fusion. AB - We examined the utility of anterior decompression and bony fusion via the extrapleural approach in the treatment of thoracic myelopathy secondary to ossification of the posterior longitudinal ligament (OPLL). Patient outcome and complications were analyzed in 48 patients treated with this procedure, with a follow-up of at least 2 years. The Japanese Orthopaedic Association score was used to evaluate the severity of the thoracic myelopathy, and the recovery rate was used to evaluate the surgical outcome. The outcome, postoperative complications, radiographic evaluations of bony union, and progression of OPLL within the area of anterior decompression were examined. The T3 vertebral body was the highest level to which anterior decompression was applied. The average follow-up period was 57 months with a recovery rate of 56.7% which stabilized 1 year after operation. However, the surgical outcome was less favorable in patients with long-standing myelopathy, extensive OPLL, or thoracic OPLL with coexisting intraspinal ligament ossification. Four patients experienced deterioration of their myelopathy, and seven patients had the postoperative complication of extraspinal leakage of cerebrospinal fluid. The myelopathy was transient in all but one patient. Radiographic studies showed that bony union was achieved and restenosis of the spinal canal due to progression of OPLL within the area of decompression did not occur. We conclude that anterior decompression and bony fusion using the extrapleural approach provides a good outcome and is useful in treating mid- and lower thoracic OPLL when performed carefully at an early stage of disease. PMID- 9392052 TI - Interactions between stone-forming calcific crystals and macromolecules. PMID- 9392053 TI - Induction of ICAM-1, HLA-DR molecules by IFN-gamma and oncogene expression in human bladder cancer cell lines. AB - It is well known that the expression level of the ICAM-1 and MHC is frequently altered in accordance with tumor progression, and the expression of oncogenes is significantly related to tumorigenesis, tumor progression, and/or metastatic potential. In this study, to investigate the relationship between the alteration in ICAM-1 and MHC expression with the tumor grade and/or cell differentiation, we examined the expression of ICAM-1 and HLA-DR before and after treatment with IFN gamma in 4 human bladder cancer cell lines. We also analyzed the expression of c Ha-ras and c-myc. Using flow cytometry, highly constitutive expression of ICAM-1 was detected in all cell lines tested except RT4 (grade I, well-differentiated, superficial). IFN-gamma was found to somewhat induce the expression of ICAM-1 in all cell lines except RT4. The constitutive expression of HLA-DR was not detected in any of the cell lines tested by flow cytometry and Northern blot. HLA-DR expression was induced by IFN-gamma in RT4 and J82 (grade III, anaplastic, invasive). Codon 12 point mutation of c-Ha-ras (GGC-->GTC; Gly-->Val) was detected in T24 (grade III, epidermoid, superficial) through single-strand conformation polymorphism, and sequencing analysis. Another point mutation at codon 27 was detected in TCCSUP (grade IV, distant metastatic), but this mutation was found to be silent (CAT-->CAC; His). Expression of c-myc was detected in J82 and TCCSUP by Northern blot. These findings, along with the clinical and pathological characteristics of the patients from whom the cell lines were established, might suggest that the expression of ICAM-1 seems to be associated with cell differentiation, and the inducibility of HLA-DR by IFN-gamma seems to be associated with the degree of malignancy. Expression of c-myc seems to be associated with invasiveness. However, a significant correlation between c-Ha-ras activation and tumor grade could not be observed. PMID- 9392054 TI - Immunohistochemical studies of proliferating cell nuclear antigen and cathepsin D in transitional cell carcinoma of the urinary bladder. AB - Immunohistochemical staining for proliferating cell nuclear antigen (PCNA) and cathepsin D was performed on 60 transitional cell carcinoma (TCC) specimens from 60 patients with bladder cancer. The percentage of PCNA-positive cells (PCNA labelling index) was determined by counting 500 or 1,000 cells, and cathepsin D expression was graded according to the extent of immunoreactivity to anti cathepsin D antibody. The PCNA-labelling index was significantly higher in high grade and high-stage tumors compared to that in low-grade and low-stage tumors. Cathepsin D was highly positive in grade-1 tumors. In contrast, 82% of grade-3 tumors and 76% of advanced tumors showed negative or low reactivity to anti cathepsin D. Groups of high PCNA-labelling index and negative cathepsin D had significantly poorer prognoses compared to those of the low PCNA group and cathepsin D highly positive group, respectively, in univariate analyses. However, neither of these two factors were independent prognostic factors in multivariate analyses. These results suggest that the PCNA-labelling index and cathepsin D expression may indicate the malignant potential of TCC and may be able to provide additional information for predicting survival when stratifying for grade of bladder cancer. PMID- 9392055 TI - Expression of immunohistochemical markers (PCNA, Ki-67, 486p and p53) on paraffin sections and their relation to the recurrence rate of superficial bladder tumors. AB - We present a retrospective study using four different immunohistochemical markers (PCNA, Ki-67, 486p and p53) on paraffin sections from 104 selected cases with primary superficial transitional cell carcinomas of the bladder (59 cases pTa, 45 cases pT1, 40 cases G1, 64 cases G2). 53 of the 104 patients experienced recurrence of their bladder lesion, while 51 remained free of tumor. The distribution of staging, grading and multifocality was comparable in both groups of patients. Overall, the tumors that recurred had a significantly higher proportion of labeled cells for PCNA (p < or = 0.0001), Ki-67 (p < or = 0.006) and 486p (p < or = 0.0001). The latter antigen proved to be the most reliable marker. A less significant difference in staining pattern was found for p53 (p < or = 0.01). Evaluating the predictive value of the various antibodies separately for the groups with G1 vs. G2 carcinomas and pTa vs. pT1 tumors revealed a lower significance for all antibodies. The technique of immunostaining on paraffin sections facilitates further retrospective studies on archival material. These markers may provide additional information about the probability of recurrence of superficial bladder tumors. But at the moment they should only be utilized in selected cases. PMID- 9392056 TI - Inhibitory effect of capsaicin on detrusor contractility: further study in the presence of ganglionic blocker and neurokinin receptor antagonist in the rat urinary bladder. AB - In order to understand the mechanisms by which capsaicin at high concentrations affects the micturition reflex and detrusor contractility, in vivo and in vitro whole bladder studies were conducted using ganglionic blockers and a neurokinin receptor antagonist. Thirty-eight adult rats were divided into control (normal saline cystometry) and experimental (1,000 microM capsaicin cystometry) groups. Both groups were subdivided to receive pretreatment with intravesical hexamethonium, perivesical hexamethonium, or intravesical spantide ([D-Arg1, D Trp7,9, Leu11]-substance P). After in vivo cystometry, the bladders were removed and in vitro whole bladder contractility studies using electrical field stimulation as well as bethanechol and KCl stimulations were performed. In the bladders pretreated with perivesical hexamethonium, the amplitudes of contractions and in vitro detrusor contractility under electrical stimulation were decreased. Other bladder preparations showed no significant differences from the controls. However, when 1,000 microM capsaicin was infused into the bladders, both control and experimental bladders showed an initial excitation and a final inhibition with an elevated basal intravesical pressure and retention. Capsaicin at 100 microM did not have this effect. The results of this study conclude that blockage of perivesical ganglia or neurokinin receptors in the submucosa did not influence the depressant effects of 1,000 microM capsaicin on the micturition reflex and detrusor contractility in rats. Nonspecific toxic effects on detrusor muscle or nerves is likely when intravesical high-concentration capsaicin is administered. PMID- 9392057 TI - Indications for open stone removal of urinary calculi. AB - In a retrospective study we analyzed patients undergoing open stone removal in the Department of Urology of the University of Tubingen. In 2.7% of all urinary calculi, open stone surgery was necessary. Open operation was performed on all patients with complete staghorn calculi as well as on patients with renal pelvic stones and simultaneous morphological obstruction. Partial staghorn calculi were operated on only after endoscopic treatment had failed. Small renal pelvic stones and ureteral stones were surgically removed only after extracorporeal shock wave lithotripsy (ESWL) and endoscopic surgery had been unsuccessful. The treatment of choice for ureteral calculi is ESWL. If ESWL is impossible, an endoscopic approach is advisable. Open operations of ureteral calculi only have to be performed if endoscopic therapy has failed or if there is a simultaneous morphological obstruction. Meta-analysis of publications from 1981 to 1995 confirmed our approach regarding indications for open stone removal. Comparison of the results reported in the literature is very difficult because of the missing, but generally accepted definition of stone free. In addition different examination techniques to determine the status 'stone free' make it difficult to compare the various studies. PMID- 9392058 TI - Is deep dorsal vein arterialization an alternative surgical approach to treat venogenic impotence? AB - OBJECTIVE: Several surgical techniques have been applied for the treatment of cavernosal venous leakage, sufficient enough to create erectile dysfunction. In light of the variable success rates, we report our experience with deep dorsal penile vein arterialization for the management of severe cavernosal venous leakage. PATIENTS AND METHODS: Twenty-four impotent patients with venous leakage were treated by the Virag-II type of end-to-end anastomosis of the inferior epigastric artery to the deep dorsal penile vein and ligation of the vein proximally. Diagnostic evaluation included color flow Doppler sonography before and after intracavernous 60 mg papaverine injection and dynamic cavernosometry/ cavernosography, which indicated veno-occlusive dysfunction in 28 patients. Revascularization was technically possible in 24 of the patients, although in 4 anastomosis could not be achieved due to the poor quality of the inferior epigastric artery. RESULTS: Five patients had early occlusion in the immediate postoperative period and 4 had late occlusion within 8 months. Potency was improved initially in 9 (38%) of the 24 patients in whom successful anastomosis had been achieved, with longer term improvement in 6 (25%) of 24 who realized restoration of erectile potency as defined by clinical investigations. The mean follow-up was 24 months (range 3-36 months). CONCLUSION: We believe that anastomosis of the inferior epigastric artery to the deep dorsal penile vein and ligation of the vein proximally in cases of venous leakage results in a low success rate due probably to a pancavernosal alteration in corporal tissue compliance. PMID- 9392060 TI - Symptomatic adrenal myelolipoma. Clinicopathological analysis of 7 cases and brief review of the literature. AB - Seven cases of adrenal myelolipoma comprising 5.8% of total adrenal tumors are described with a male-to-female ratio of 1.3:1. Five were symptomatic, of which 4 had a palpable abdominal mass, 2 cases detected incidentally were associated with carcinoma of the uterine cervix and renal cell carcinoma, respectively. The average age and duration of symptoms at presentation were 56 years (range 38-70 years) and 3.16 months (range 1-9 months). A CT scan was done in all cases, of which 5 showed a nonenhancing mass lesion with fat density diagnosed as adrenal myelolipoma. However, in 1 case radiological diagnosis of liposarcoma was maintained because of the huge size of the lesion whereas in another case the lesion was missed because of associated renal cell carcinoma. Interestingly the right adrenal was involved in all cases and the weight varied from 7 to 2,000 g. No recurrence was noted in the follow-up period (ranging from 3 months to 10 years). PMID- 9392059 TI - Right colonic reservoir with submucosally embedded tapered ileum--'Tiflis pouch'. AB - Seventeen patients with invasive bladder cancer aged between 44 and 67 years underwent radical cystectomy and construction of the continent urinary reservoir from 20 to 25 cm cecoascendum reconfigurated a.m. Heineke-Mikulicz. and submucosally embedded tapered ileum as a continence mechanism--'Tiflis pouch'. There were no perioperative mortalities and only minor complication but abdominal wound dehiscence occurred with a subsequent hernia in 1 obese patient. One patient died of urethral recurrence and malignancy progression 14 months after surgery. During the follow-up period of 9-37 months, all renal units remained unobstructed. Eleven of 15 preoperatively dilated units improved. No case of pouch stone formation, stomal stenosis or difficulties with catheterization was observed. All patients are continent but 4 need catheterization at 3-hour intervals. The functional capacity is in the range of 310-560 ml. The Tiflis pouch is a capacious reservoir in which the continence state is satisfactory, perioperative complication rate is low and quality of life is high. PMID- 9392061 TI - Hemangioma of the bladder with extravesical extension. AB - Bladder hemangioma is a rare benign tumor occurring in any age group. We report the case of a 76-year-old woman having a bladder hemangioma. On cystoscopic examination no typical features of the bladder hemangioma were found. Biopsy of the lesion revealed a cavernous hemangioma. A partial cystectomy was performed for the control of macroscopic hematuria. PMID- 9392062 TI - An unusual complication following uroflowmetry: water intoxication resulting in hyponatremia and seizure. AB - Uroflowmetry is considered a simple and noninvasive test in the evaluation of urinary symptoms. It requires patients to consume fluid orally for a full bladder prior to undertaking the test. Guidelines regarding the amount and rate of oral fluid intake have not been accurately defined. We report on a patient who suffered a serious complication of water intoxication with hyponatremia and seizure due to excessive water consumption and absorption during uroflowmetry. We discuss the underlying factors concerning this complication and recommend a more conservative approach to attain a full bladder in a certain subgroup of patients at risk of developing such a complication. PMID- 9392063 TI - Influence of the force applied and its period of application on the outcome of the flexion test of the distal forelimb of the horse. AB - The influence of the force applied and its period of application on the outcome of the flexion test of the distal forelimb was investigated in a group of eight sound horses. The degree of lameness after the flexion test was scored by a standard clinical classification, and by measuring the angle of maximum fetlock extension by means of the infrared light-based MacReflex gait analysis system. There was a good correlation between the clinical score and this electronically recorded kinematic parameter (r = 0.96). Both the force applied and the period of application affected the outcome of the test. Increasing the force applied by 25 per cent led to three horses being judged positive, instead of two when the normal force was applied. Doubling the time to 120 seconds resulted in four horses rather than two being classified as lame after the test. Reducing the force to 75 per cent or the time to 30 seconds resulted in all the horses being classified as sound. A flexion test lasting five minutes, either at 100 per cent force or at 75 per cent, classified six of the eight horses as lame. It is concluded that the flexion test should be defined more precisely in terms of these two factors in order to make its results more consistent and hence more useful. PMID- 9392064 TI - Omasal and abomasal impaction in beef suckler cows. AB - A group of late gestation suckler cows were housed in straw yards and had been fed solely on pea haulm for the previous three weeks. Four cows became ill, with a variety of clinical signs, two died, one was euthanased and one recovered spontaneously. Postmortem examination revealed severe omasal and abomasal impaction. No further cases occurred after changes were made to the diet of the cows. PMID- 9392065 TI - Immunotoxicological effects on piglets of feeding sows diets containing aflatoxins. AB - Three groups of four Large White sows were fed diets containing either 800 ppb purified aflatoxin B1 (group 1), 800 ppb purified aflatoxin G1 (group 2) or 400 ppb B1 and 400 ppb G1 (group 3) throughout gestation and lactation. A control group of four sows was fed a diet free of aflatoxins. Aflatoxins B1 and M1 were found in milk samples taken five and 25 days after parturition from the sows of group 1, aflatoxin G1 was present in the milk of the sows of group 2 and all three aflatoxins were present in samples from the sows of group 3. The concentration of aflatoxin in the milk was about 1000-fold lower than that in the feed, but increased over the 25 days after parturition. The piglet suckling on a central teat was selected from each sow, given sow milk until the fourth day of age, and was then free to eat prepared feed while suckling. At the 25th day of age the selected piglets were removed from the sow and sacrificed. Blood samples were collected from each piglet and cellular populations were separated for immunological measurements: an in vitro lymphocyte proliferation test, and tests to derive the phagocytic activity, phagocytic index and superoxide anion production of monocyte-derived macrophages were carried out along with studies on the motility, differential chemotaxis and chemotactic index of circulating granulocytes. The lymphoproliferative response to mitogens was reduced and monocyte-derived macrophages failed to efficiently produce superoxide anions after oxidative burst stimulation in vitro, while their ability to phagocytose red blood cells was not compromised. Granulocytic cells showed a reduction of chemotactic response in vitro to chemoattractant bacteria factor and casein. PMID- 9392066 TI - Use of arithmetic and geometric means in the calculation of anthelmintic efficacy. PMID- 9392067 TI - Cutaneous lymphoma in a Scottish blackface ram. PMID- 9392068 TI - Endogenous lipid pneumonia (alveolar histiocytosis) and hydrocephalus in an adult llama (Llama glama). PMID- 9392069 TI - Vaccination of farmed crocodiles (Crocodylus niloticus) against Mycoplasma crocodyli infection. PMID- 9392070 TI - Generalised caseous lymphadenitis. PMID- 9392071 TI - Porcine dermatitis and nephropathy syndrome in the USA. PMID- 9392072 TI - Therapeutic claims for unlicensed products. PMID- 9392073 TI - Ragwort poisoning in a pedigree cow. PMID- 9392074 TI - The homeodomain protein Pho2p binds at an A/T-rich segment flanking the binding site of the basic-helix-loop-helix protein Pho4p in the yeast PHO promoters. AB - Transcription of the genomic PHO5, PHO81 and PHO84 genes of the PHO regulon requires Pho4p and Pho2p activity, whereas transcription of PHO8 is directed by Pho4p alone. Pho4p binds to two 9-bp motifs, 5'-GCACGTGGG-3' (type 1. e.g. UASp2 of PHO5 and site D of PHO84) and 5'-GCACGTTTT-3' (type 2, e.g. UASp1 of PHO5 and site E of PHO84) in the PHO promoter. Experiments were performed to evaluate the ability of these 9-bp motifs to function as upstream activation sites (UASs) by insertion of various 36-bp fragments bearing the 9-bp motif in a CYC1-lacZ fusion gene. No expression of the lacZ gene was detected with the 36-bp fragment bearing UASp2 of PHO5, whereas similar 36-bp fragments bearing UASp1 of PHO5 and sites D and E of PHO84 showed UAS activity in response to Pi concentration in the medium and to the pho2 mutation. The Pho2p-responsive UASs are flanked by one or two copies of an A/T-rich segment, whereas UASp2 is not. Gel retardation and competition experiments performed using a T7-Pho2p-His chimeric protein showed that Pho2p binds to the 36-bp fragments bearing A/T-rich segment(s) but not appreciably to the 36-bp fragments not bearing such segment(s). Thus, the A/T segments flanking the PHO UASs are Pho2p binding sites and play an important role in PHO regulation. PMID- 9392075 TI - Purification and characterization of phosphofructokinase from the yeast Kluyveromyces lactis. AB - Phosphofructokinase from Kluyveromyces lactis was purified by 180-fold enrichment, elaborating the following steps: cell disruption, polyethylene glycol precipitation, affinity chromatography, size exclusion chromatography on Sepharose 6B and on Bio-Sil SEC 400 and ion exchange chromatography. The homogeneous enzyme exhibits a molecular mass of 845 +/- 20 kDa as determined by sedimentation equilibrium measurements and a specific activity of 100 units/mg protein. The apparent sedimentation coefficient was found to be s20,c = 20.7 +/- 0.6 S and no significant dependence on the protein concentration was observed in a range from 0.2 to 8 mg protein/ml. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed two bands corresponding to molecular masses of 119 +/- 5 kDa and 102 +/- 5 kDa, respectively. Thus, the enzyme assembles as octamer composed of two types of subunits. From Western blot analysis applying subunit specific monoclonal antibodies raised against Saccharomyces cerevisiae phosphofructokinase and from the determination of the N-terminal amino acid sequence, the conclusion was drawn that the 102 kDa-subunit corresponds to the beta-subunit of the S. cerevisiae enzyme. In contrast to bakers' yeast phosphofructokinase, the K. lactis enzyme exhibits no cooperativity with respect to the substrate fructose 6-phosphate. Both activators AMP and fructose 2,6 bisphosphate decrease the Michaelis constant with respect to this substrate. The enzyme from K. lactis is also inhibited by ATP. Fructose 2,6-bisphosphate or AMP diminish the ATP-inhibition. In contrast to the phosphofructokinase from S. cerevisiae, where fructose 2,6-bisphosphate turned out to be more efficient than AMP, both activators exert similar effects on the K. lactis enzyme. PMID- 9392076 TI - The ALD6 gene of Saccharomyces cerevisiae encodes a cytosolic, Mg(2+)-activated acetaldehyde dehydrogenase. AB - The deduced translation product of an open reading frame on the left arm of chromosome XVI of Saccharomyces cerevisiae, with the systematic name of YPL061w, is 500 amino acids in length and shares significant homology with aldehyde dehydrogenases. Amino acids 2 to 16 of the protein encoded by YPL061w were found to be identical to the N-terminal 15 amino acids of the purified cytosolic, Mg(2+)-activated acetaldehyde dehydrogenase (ACDH) of S. cerevisiae. This enzyme is thought to be involved in the production of acetate from which cytosolic acetyl-CoA is then synthesized. Deletion of YPL061w was detrimental to the growth of haploid strains of yeast; an analysis of one deletion mutant revealed a maximum specific growth rate (in complex medium containing glucose) of one-third of that displayed by the wild-type strain. Mutants deleted in YPL061w were also unable to use ethanol as a carbon source. As expected, the cytosolic, Mg(2+) activated ACDH activity had been lost from the mutants, although the mitochondrial, K(+)-activated ACDH was readily detected. PMID- 9392077 TI - Measurement of nuclear DNA content in fission yeast by flow cytometry. AB - Cell division cycle (cdc) mutants of Schizosaccharomyces pombe are arrested at specific points in the cell cycle when grown at restrictive temperature. Flow cytometry of such cells reveals an anomalous increase in the DNA fluorescence signal, which represents a problem in experiments designed to determine the cell cycle arrest point. The increased fluorescence signal is due to cytoplasmic constituents and has been attributed to mitochondrial DNA synthesis (S. Sazer and S. W. Sherwood, J. Cell Sci. 97: 509-516, 1990). Here we have studied the cdc10 mutant by flow cytometry using different DNA-binding fluorochromes and found no evidence that the increased fluorescence signal was caused by mitochondrial DNA synthesis. To determine more accurately the nuclear DNA content we have developed a novel method to remove most of the cytoplasmic material by exposing the cells to Triton X-100 and hypotonic conditions after cell wall digestion. The DNA fluorescence from cells treated in this way was more constant with time of incubation at restrictive temperature in spite of a considerable increase in cell size. With this method we could determine that the recently isolated temperature sensitive orp1 mutant is arrested with a 1C DNA content. Premature and abnormal mitosis ('cut') could be observed for the orp1 mutant after only 4 h at restrictive temperature. PMID- 9392078 TI - Lys80p of Saccharomyces cerevisiae, previously proposed as a specific repressor of LYS genes, is a pleiotropic regulatory factor identical to Mks1p. AB - In Saccharomyces cerevisiae, an intermediate of the lysine pathway, alpha aminoadipate semialdehyde (alpha AASA), acts as a coinducer for the transcriptional activation of LYS genes by Lys14p. The limitation of the production of this intermediate through feedback inhibition of the first step of the pathway results in apparent repression by lysine. Previously, the lys80 mutations, reducing the lysine repression and increasing the production of lysine, were interpreted as impairing a repressor of LYS genes expression. In order to understand the role of Lys80p in the control of the lysine pathway, we have analysed the effects of mutations epistatic to lys80 mutations. The effects of lys80 mutations on LYS genes expression were dependent on the integrity of the activation system (Lys14p and alpha AASA). The increased production of lysine in lys80 mutants appeared to result from an improvement of the metabolic flux through the pathway and was correlated to an increase of the alpha-ketoglutarate pool and of the level of several enzymes of the tricarboxylic acid cycle. The LYS80 genes has been cloned and sequenced; it turned out to be identical to gene MKS1 cloned as a gene encoding a negative regulator of the RAS-cAMP pathway. We conclude that Lys80p is a pleiotropic regulatory factor rather than a specific repressor of LYS genes. PMID- 9392079 TI - A rapid and reliable method for metabolite extraction in yeast using boiling buffered ethanol. AB - A simple and reliable method for the efficient inactivation of metabolism and for quantitative metabolite extraction from yeast cells is presented. It is based on the use of a boiling solution made of 75% ethanol (volume/final volume) buffered with 70 mM-Hepes (final concentration), pH 7.5, to guarantee the stability throughout the whole procedure of a large variety of metabolites, including all glycolytic intermediates, nucleotides, pyridine nucleotides and organic acids compounds. The extraction is fast, requiring only 3 min incubation of yeast cells in the ethanol-buffered mixture maintained at 80 degrees C. It can be carried out either directly by spraying the cells into the boiling mixture, or after quenching the whole culture in 60% methanol kept at -40 degrees C. Extracts are subsequently concentrated by evaporation under partial vacuum and the residue is resuspended in a small volume of water. This concentration step and the use of a highly sensitive analytical method allow us to quantify metabolites in less than 10 mg dry weight cells. This method, which can be applied to other fungi, could be very helpful for the determination of true metabolites in mutants generated through the EUROFAN programme and for metabolic flux analysis. PMID- 9392080 TI - Functional differences among the six Saccharomyces cerevisiae tRNATrp genes. AB - The Saccharomyces cerevisiae haploid genome includes six copies of the gene encoding tRNATrp which are scattered on five chromosomes. Other, non-functional tDNATrp fragments also occur in the genome. The segments of all six genes which encode the 72-nucleotide mature tRNATrp, as well as a 34-nucleotide intervening sequence, are identical. However, the 5' and 3' flanking sequences diverge virtually at the boundaries of the coding region. We have used an assay based on suppression of UGA mutations by multi-copy clones of tDNATrp to search for functional differences among these genes. Previous studies with one tDNATrp had demonstrated that moderate suppression of a UGA mutation, leu2-2, resulted from introduction of a multi-copy clone of the gene. Attempts to use this assay to select tDNATrp clones from a yeast genomic library yielded only four of the six different clones. The other two genes were amplified by PCR and cloned in pRS202, a 2 mu vector also used for the genomic library. Plasmids bearing the six tRNA genes were transformed into S. cerevisiae strain JG369.3B and scored for their ability to suppress the leu2-2 mutation as well as his4-260, another UGA marker. Two of the six tRNATrp clones were unable to suppress either marker, two evidenced weak suppression of the Leu auxotrophy, and two were able to suppress both markers. Growth rates in liquid media requiring suppression were measured for cell lines carrying each of the clones. Differences greater than 50-fold were observed in media lacking histidine. An evolutionary tree based on 5'-flanking sequence corresponds reasonably well with suppressor activity, while a similar analysis of 3'-flanking sequence does not. This suggests that the functional differences are based on divergence in the 5'-flanking sequences of the tRNATrp genes. PMID- 9392081 TI - Characterization of new proteins found by analysis of short open reading frames from the full yeast genome. AB - We have analysed short open reading frames (between 150 and 300 base pairs long) of the yeast genome (Saccharomyces cerevisiae) with a two-step strategy. The first step selects a candidate set of open reading frames from the DNA sequence based on statistical evaluation of DNA and protein sequence properties. The second step filters the candidate set by selecting open reading frames with high similarity to other known sequences (from any organism). As a result, we report ten new predicted proteins not present in the current sequence databases. These include a new alcohol dehydrogenase, a protein probably related to the cell cycle, as well as a homolog of the prokaryotic ribosomal protein L36 likely to be a mitochondrial ribosomal protein coded in the nuclear genome. We conclude that the analysis of short open reading frames leads to biologically interesting discoveries, even though the quantitative yield of new proteins is relatively low. PMID- 9392082 TI - Isolation of a putative prolyl-tRNA synthetase (CaPRS) gene from Candida albicans. AB - We have isolated a 4.0-kb fragment from a genomic library of Candida albicans which contained two open reading frames (ORFs). One of them is homologous to a prolyl-tRNA synthetase that catalyses the charging of a specific tRNA by proline (CaPRS). A deduced sequence of 575 amino acids representing a polypeptide of 66.2 kDa was determined. A FASTA search indicated that the CaPRSp had an overall similarity of 54.4% with the product of a Saccharomyces cerevisiae ORF (YER087) and 43.8% with the prolyl-tRNA synthetase of Escherichia coli (COLIPRO). Consensus Class II aminoacyl-tRNA synthetase sequences were identified by the PROSITE program. CaPRS was localized to chromosome R of the C. albicans genome and CaPRS DNA hybridized to a major RNA transcript of 1.7 kb under all conditions tested. PMID- 9392083 TI - Isolation and sequence of the gene encoding ornithine decarboxylase, SPE1, from Candida albicans by complementation of a spe1 delta strain of Saccharomyces cerevisiae. AB - The gene encoding ornithine decarboxylase, SPE1, from the pathogenic yeast Candida albicans has been isolated by complementation of an ornithine decarboxylase-negative (spe1 delta) strain of Saccharomyces cerevisiae. Four transformants, three of which contain plasmids with the SPE1 gene, were isolated by selection on polyamine-free medium. The C. albicans ornithine decarboxylase (ODC) showed high homology with other eukaryotic ODCs at both the amino acid and nucleic acid levels. PMID- 9392084 TI - Current awareness on yeast. PMID- 9392086 TI - [Carbohydrates and the human being]. PMID- 9392087 TI - [Glycogen synthesis and glycogenolysis]. PMID- 9392085 TI - [Several problems in the clinical aspects of diabetes mellitus]. PMID- 9392088 TI - [Gluconeogenesis and glycolysis]. PMID- 9392089 TI - [TCA cycle and electron transport system]. PMID- 9392090 TI - [Glucose transporter]. PMID- 9392092 TI - [Glucose transport in the kidney]. PMID- 9392091 TI - [Mechanisms of carbohydrate digestion and absorption in the intestinal tract]. PMID- 9392093 TI - [Review: regulatory factors of glucose metabolism]. PMID- 9392094 TI - [Insulin gene: gene organization and its regulatory mechanism of transcription]. PMID- 9392095 TI - [Insulin biosynthesis and its chemical structure]. PMID- 9392096 TI - [Molecular mechanism of insulin secretion]. PMID- 9392097 TI - [Biphasic insulin secretion and glucose metabolism]. PMID- 9392098 TI - [The structure and the regulation of the gene expression of human insulin receptor gene]. PMID- 9392099 TI - [Structure and biosynthesis of human insulin receptor]. PMID- 9392100 TI - [Mechanism of insulin actions]. PMID- 9392101 TI - [Insulin, its kinetics and metabolism]. PMID- 9392102 TI - [Glucagon, GLP]. PMID- 9392103 TI - [Somatostatin]. PMID- 9392104 TI - [Gastrointestinal hormones in diabetes mellitus]. PMID- 9392105 TI - [Anterior pituitary hormones--mainly about growth hormone]. PMID- 9392106 TI - [Biogenic amines]. PMID- 9392107 TI - [Glucocorticoid]. PMID- 9392108 TI - [Regulation of glucose metabolism by thyroid hormones]. PMID- 9392110 TI - [Glucose, protein, and lipid metabolism and their regulation system in adipocyte]. PMID- 9392109 TI - [Hepatocyte]. PMID- 9392111 TI - [Specific glucose metabolism in muscle]. PMID- 9392112 TI - [Brain energy metabolism]. PMID- 9392113 TI - [Vascular endothelial cells]. PMID- 9392114 TI - [Intestinal epithelial cells]. PMID- 9392115 TI - [Renal tubular cells]. PMID- 9392116 TI - [Definition and classification of diabetes mellitus]. PMID- 9392117 TI - [Problems on the diagnostic criteria for diabetes mellitus]. PMID- 9392118 TI - [Early diagnosis of NIDDM--theory and direction]. PMID- 9392119 TI - [Natural history of diabetes mellitus]. PMID- 9392120 TI - [IDDM--overview]. PMID- 9392121 TI - [Susceptibility genes for IDDM]. PMID- 9392122 TI - [Molecular mechanisms of pancreatic beta-cell destruction in IDDM]. PMID- 9392123 TI - [Destruction and regeneration of pancreatic beta cells]. PMID- 9392124 TI - [Adhesion molecules and diabetes mellitus]. PMID- 9392125 TI - [General aspects of etiology and pathogenesis of NIDDM]. PMID- 9392126 TI - [Molecular genetics of NIDDM]. PMID- 9392127 TI - [Molecular mechanisms for impaired insulin secretion in NIDDM]. PMID- 9392128 TI - [Defects in insulin's signalling process and diabetes mellitus]. PMID- 9392129 TI - [Primary, secondary, tertiary prevention of diabetes mellitus]. PMID- 9392130 TI - [Lifestyle interventions for preventing non insulin-dependent diabetes mellitus]. PMID- 9392131 TI - [Possible primary prevention of NIDDM (type 2 diabetes) by pharmacological interventions]. PMID- 9392133 TI - [Association of HLA genes with IDDM: a review]. PMID- 9392132 TI - [Gene mutations in diabetes mellitus]. PMID- 9392134 TI - [IDDM and variable number of tandem repeats (VNTR) in the 5'region of the insulin gene: a review]. PMID- 9392135 TI - [TNF gene polymorphism and IDDM]. PMID- 9392136 TI - [Glutamic acid decarboxylase gene]. PMID- 9392137 TI - [Type 1 diabetes susceptibility genes on human chromosome 2q]. PMID- 9392138 TI - [Insulin gene]. PMID- 9392139 TI - [Insulin receptor gene]. PMID- 9392140 TI - [Insulin promoter factor-1]. PMID- 9392141 TI - [Glucokinase gene and its dual promoter regions]. PMID- 9392142 TI - [Insulin receptor substrate-1 (IRS-1) gene]. PMID- 9392143 TI - [Glucagon receptor gene]. PMID- 9392144 TI - [Glucagon-like peptide-1 (GLP-1) receptor gene]. PMID- 9392145 TI - [Fatty acid binding protein-2(FABP-2) gene]. PMID- 9392146 TI - [Glycogen synthase gene]. PMID- 9392147 TI - [Amylin (IAPP) gene]. PMID- 9392148 TI - [ATP sensitive potassium channels--Kir6.2 gene]. PMID- 9392149 TI - [ATP-sensitive potassium channel--sulfonylurea receptor]. PMID- 9392150 TI - [Proinsulin converting enzyme (PC2, PC3) gene]. PMID- 9392151 TI - [Beta 3-adrenergic receptors]. PMID- 9392152 TI - [Glucose transporter gene]. PMID- 9392153 TI - [Hexokinase gene]. PMID- 9392155 TI - [Apolipoprotein genes]. PMID- 9392154 TI - [ob gene (leptin gene)]. PMID- 9392156 TI - [Mitochondria glycerol-3-phosphate dehydrogenase gene]. PMID- 9392157 TI - [Mutation/polymorphism of phosphofructokinase (PFK) genes in diabetes mellitus]. PMID- 9392158 TI - [Islet 1 gene]. PMID- 9392159 TI - [Gastric inhibitory polypeptide (GIP) receptor]. PMID- 9392160 TI - [Secondary forms of diabetes]. PMID- 9392161 TI - [Pathology of diabetes mellitus]. PMID- 9392162 TI - [Epidemiology of diabetes mellitus in Japan]. PMID- 9392163 TI - [Prognosis and causes of death in patients with non-insulin-dependent diabetes mellitus (NIDDM)]. PMID- 9392164 TI - [Epidemiology of diabetes mellitus in the elderly]. PMID- 9392165 TI - [Epidemiology of diabetes mellitus in the world except Japan]. PMID- 9392166 TI - [Family history of diabetes]. PMID- 9392167 TI - [Symptomatology of diabetes mellitus]. PMID- 9392168 TI - [Abnormal carbohydrate metabolism in diabetes mellitus]. PMID- 9392169 TI - [Abnormalities in lipid metabolism associated with diabetes mellitus]. PMID- 9392170 TI - [Lipoprotein abnormalities in normolipidemic diabetics]. PMID- 9392171 TI - [Diabetes-induced abnormality in protein metabolism]. PMID- 9392172 TI - [Abnormalities of amino acid metabolism in diabetes mellitus]. PMID- 9392173 TI - [Metabolic disorders of organic acids in diabetes]. PMID- 9392174 TI - [Altered uric acid metabolism in diabetes mellitus]. PMID- 9392175 TI - [Diabetes and vitamin deficiency]. PMID- 9392176 TI - [Water and electrolyte disorders and acid-base imbalance in diabetic patients]. PMID- 9392177 TI - [Disturbance of mineral metabolism]. PMID- 9392178 TI - [Trace elements in diabetes mellitus]. PMID- 9392179 TI - [Endocrine disorders in diabetes mellitus]. PMID- 9392180 TI - [Platelet dysfunction]. PMID- 9392181 TI - [Abnormality of clotting and fibrinolysis in diabetes mellitus]. PMID- 9392182 TI - [Hemorheological abnormality in diabetes mellitus]. PMID- 9392183 TI - [Diabetes mellitus and associated hypertension]. PMID- 9392184 TI - [Immunological disorder in diabetes mellitus]. PMID- 9392185 TI - [Abnormal arachidonate metabolites in diabetes mellitus]. PMID- 9392186 TI - [Abnormalities in inositol phospholipid metabolism]. PMID- 9392187 TI - [Diabetic complications--definition, classification, epidemiology]. PMID- 9392188 TI - [Risk factors for the development of diabetic complications]. PMID- 9392189 TI - [Molecular biology of atherosclerosis]. PMID- 9392190 TI - [Vascular endothelial cell injury in diabetes mellitus]. PMID- 9392191 TI - [Metabolic disorder of polyol pathway]. PMID- 9392192 TI - [Oxidative stress and atherosclerosis]. PMID- 9392193 TI - [Diabetes mellitus and cerebrovascular disorders]. PMID- 9392194 TI - [Coronary artery disease, heart failure and cardiomyopathy in diabetic patients]. PMID- 9392195 TI - [Arteriosclerosis obliterans in diabetes mellitus]. PMID- 9392196 TI - [Diabetic nephropathy--definition, stages, measurement of albuminuria, and therapy]. PMID- 9392197 TI - [Pathology of diabetic nephropathy]. PMID- 9392198 TI - [Diabetic nephropathy--risk factors and incidences]. PMID- 9392199 TI - [Familial clustering and genetics of diabetic nephropathy]. PMID- 9392200 TI - [Diabetic nephropathy--predictive methods]. PMID- 9392201 TI - [The effect of nutritional arrangement and glycemic control on the progression of chronic renal failure in diabetic patients]. PMID- 9392202 TI - [Renal replacement therapy for diabetic nephropathy]. PMID- 9392203 TI - [Survival of patients with diabetic nephropathy on hemodialysis or CAPD]. PMID- 9392204 TI - [Diabetic neuropathy--concept, classification, diagnosis, treatment]. PMID- 9392205 TI - [Diabetic neuropathy--its pathology]. PMID- 9392206 TI - [Diabetic neuropathy--risk factors and pathogenesis]. PMID- 9392207 TI - [Early detection for diabetic neuropathy]. PMID- 9392208 TI - [Electrophysiology in diabetic neurological complications]. PMID- 9392209 TI - [Sensory disorders and pain treatment in diabetic neuropathy]. PMID- 9392210 TI - [Diabetic autonomic neuropathy]. PMID- 9392211 TI - [Diabetic urinary bladder and urethral dysfunction]. PMID- 9392212 TI - [Diabetic neurogenic arthropathy (Charcot's joint)]. PMID- 9392213 TI - [Infectious complications of diabetes mellitus]. PMID- 9392214 TI - [Liver, biliary, and pancreas disease in diabetes mellitus]. PMID- 9392215 TI - [Gastrointestinal complications of diabetes mellitus]. PMID- 9392216 TI - [Diabetic gastropathy]. PMID- 9392217 TI - [Diabetic foot]. PMID- 9392218 TI - [Autoimmune disease in patients with diabetes mellitus]. PMID- 9392219 TI - [Endocrinopathy associated with diabetes mellitus]. PMID- 9392220 TI - [Diabetic sexual impotence]. PMID- 9392221 TI - [Diabetic osteopenia]. PMID- 9392222 TI - [Diabetic retinopathy--classification, diagnosis, treatment]. PMID- 9392223 TI - [Diabetic retinopathy--pathology]. PMID- 9392224 TI - [Risk factors for diabetic retinopathy]. PMID- 9392225 TI - [Molecular pathogenesis of diabetic retinopathy]. PMID- 9392226 TI - [Method of early detection at every stage of diabetic retinopathy]. PMID- 9392227 TI - [Conservative therapy of diabetic retinopathy]. PMID- 9392228 TI - [Treatment for diabetic retinopathy using photocoagulation and vitrectomy]. PMID- 9392229 TI - [The cutaneous manifestations of diabetes mellitus]. PMID- 9392230 TI - [Diabetic orthostatic hypotension]. PMID- 9392231 TI - [Primary glomerulonephritis complicating diabetes mellitus]. PMID- 9392232 TI - [Diabetes mellitus and periodontal disease]. PMID- 9392233 TI - [Diabetes mellitus and sleep apnea syndrome]. PMID- 9392234 TI - [Diabetic amyotrophy]. PMID- 9392235 TI - [Rhabdomyolysis in the diabetic state]. PMID- 9392237 TI - [Hyperosmolar nonketotic coma]. PMID- 9392236 TI - [Diabetic ketoacidotic coma]. PMID- 9392238 TI - [Lactic acidosis]. PMID- 9392239 TI - Healthcare assistants: the substitute nurse. PMID- 9392240 TI - New deal for people with learning difficulties. PMID- 9392241 TI - I.v. therapy: selection, use and management of infusion pumps. AB - A locally agreed, coordinated policy is essential for the selection and purchase of infusion pumps, and should form part of overall policy for medical device management. When developing a purchasing policy the core needs of all relevant professional groups should be addressed, to ensure that an informed, integrated strategy evolves. This will enhance the wider responsibilities associated with, for example, device training and education, and medical device maintenance, as well as the basic clinical requirements of the infusion pump. The Medical Devices Agency provides guidance on the selection and purchasing of infusion pumps through its medical device evaluation programme, which uses evidence-based systems that categorize devices by clinical infusion risk. PMID- 9392242 TI - Why should acute trusts be interested in cardiac rehabilitation? AB - This questionnaire survey of 100 ex-coronary care unit (CCU) patients in the Royal Devon and Exeter Hospital NHS Trust sought to determine patients' views (i.e. consumers' rather than professionals' views) regarding their perceived need for individual of group support following myocardial infarction, and whether the information received in hospital was adequate for their needs. A literature review of the evidence suggesting that cardiac rehabilitation is effective is included. Responses indicated that 42 of the 62 patients who responded (67.7%) felt that they would have benefited from a relaxation and/or support group, with 40 (69%) stating that they would have attended such a facility after discharge. The nature of the information offered by nursing staff while patients were in hospital was clearly that which was 'convenient' and 'unthreatening' for nurses to deliver. Only 35 respondents (56.4%) agreed that this information was adequate for their needs. These results indicate the preferences of our local consumers, whose views should be paramount in the contemporary NHS. There is a significant unmet demand for such services, which acute sector staff have the expertise to meet. PMID- 9392243 TI - Participatory research methods: people with learning difficulties. AB - The use of participatory research methods as a means of empowering disadvantaged and oppressed groups or individuals has attracted increasing interest in recent years. This article critically examines the use of such methods to empower people with learning difficulties as co-researchers. Emancipatory research would, by definition, be led and processed by people with learning difficulties. For the time being, however, the possibility of engaging people with learning difficulties in truly emancipatory nursing research is regarded as highly problematic, since it assumes empowerment as a precondition. As a step towards emancipatory research, participatory research represents a radical shift in the research process. It may potentially strengthen the voice of people with learning difficulties and enable them to express their views on nursing. The author proposes a methodology which addresses a number of critical issues facing the nurse researcher. It is a step towards developing more liberating and emancipatory methodologies. PMID- 9392244 TI - Future directions for learning disability nursing. AB - Over the past decade learning disability nurses have had to make many changes in the way they practice. Changes in attitude towards the way in which people with learning disabilities are cared for has meant that nurses have had to adjust their practice to address different expectations in the way these services are delivered. Nursing now provides opportunities for people with learning disabilities to express themselves and become involved in decision-making that affects their lives. For nurses this is a totally new way of working with this client group. Learning disability nurses have risen to the challenge that has resulted from moving into community care and forming links with primary healthcare teams. PMID- 9392245 TI - Malnutrition in hospital: detection and consequences. AB - Malnutrition in hospital is not a new concept. Studies carried out over 20 years ago identified that 50% of surgical patients (Hill et al, 1977) and 44% of medical patients (Bistrian et al, 1976) showed evidence of malnutrition. A more recent study demonstrated that the problem still exists (McWhirter and Pennington, 1994). This research showed that 40% of patients were undernourished on admission to hospital and 78% of these suffered further deterioration in their nutritional status during their hospital stay. This article, the first in the series, aims to identify patient groups which are likely to become malnourished during hospitalization and outlines how nurses and doctors can detect malnutrition and/or potential malnutrition in patients. The consequences of malnutrition and the current issues surrounding the reported problems of patients who do not receive enough food and drink while in hospital (Association of Community Health Councils, 1997) are discussed. The article also explores what measures can be taken to improve patients nutritional intake while in hospital. It describes the way forward in terms of clinical practice, current projects and the information and training available to nurses and other members of the multidisciplinary team. The author's overall aim is to improve healthcare professionals' knowledge within this field and to raise the profile of nutritional care. PMID- 9392246 TI - The abuse of specialist nursing titles must stop. PMID- 9392247 TI - Caring in the nineties. PMID- 9392248 TI - A Christian perspective of caring. PMID- 9392249 TI - Making a difference: competent to care. PMID- 9392250 TI - History in nursing. PMID- 9392251 TI - [Cesarean section]. PMID- 9392252 TI - [Approval of course]. PMID- 9392253 TI - [Women should be able to choose cesarean section themselves. Interview by Bjorg Engdahl]. PMID- 9392254 TI - [Cesarean section for the 3rd time. Interview by Bjorg Engdahl]. PMID- 9392255 TI - [Disagreement among obstetricians: one cannot squander cesarean sections. Interview by Bjorg Engdahl]. PMID- 9392256 TI - [Committed midwife becomes clinical specialist. Interview by Ingrid M Hoie]. PMID- 9392257 TI - [Many rare diagnoses collected in Sunaas. Interview by Per Flakstad]. PMID- 9392258 TI - [Health care failure--midwife's responsibility?]. PMID- 9392259 TI - Performance improvement, accreditation, and case management: defining the link. PMID- 9392260 TI - Complementary nursing--an acute care case management model. Part I--development and implementation. AB - Complementary nursing was developed in response to the need for maintaining high quality care while controlling health care costs. The complementary nurse provides comprehensive management of complex patients through an entire episode of an acute illness, to transition them back to a prehospital state through interdisciplinary discharge planning. In this article, the authors describe the process used in developing and implementing this new integrated role of acute care case management. The article contains role responsibilities, communication tools, and lessons learned from experience. PMID- 9392261 TI - The need for information-based practice in case management. AB - For case management, information-based practice is not only important but mandatory in the current competitive healthcare settings that require "cutting edge" knowledge of clinical research, business administration, managed care principles, and quality and cost-effective standards of practice. Obtaining case management-related electronic information through the Internet and CD-ROM computer databases can provide critical tools to obtain this knowledge quickly and promote higher quality practice of case management. PMID- 9392262 TI - Ethical decisions with limited resources. How is that possible? AB - Ethical decision making is a process case managers can use to help negotiate through complicated situations. In our complex health care delivery systems, case managers often have competing benefits and burdens to balance. In this article, the author presents an ethical decision making model that can assist in the process of decision making. The obligations of case managers to patients remain constant-attempting to be of benefit, but as the trajectory of care changes from acute to chronic to dying, the understanding of benefit will change. Caring for patients and their families within a managed care environment entails always keeping the benefit of the self, patient, family, employers, and society all in balance. PMID- 9392263 TI - Consistency in prevention and treatment of pressure ulcers through a wound care team. PMID- 9392264 TI - Case management for the self-insured hospital. AB - It is rare for employers who self insure to have on-site case management. Generally, self-insured employers rely on the third-party-plan administrators to control health care costs, provide enrollees with health care benefit management services, and maintain customer satisfaction. In this article, the author discusses the internal workings of an employee case management program implemented in a self-insured hospital in Northwest Indiana. Information is presented on the goals and role of the case manager, scope of practice, barriers to the program, and resources involved to deliver health care services to its enrollees. Also presented is pertinent information to track and trend data for administrative reports. In addition, customer satisfaction and the value of an on site case manager for self-insured organizations will be addressed. The results demonstrate a reduction in the volume of inpatient admissions, length of stay, and, therefore, an improved use of the health plan. PMID- 9392265 TI - Building stress resilience for nurse case managers. PMID- 9392266 TI - A case map reduces time to administration of thrombolytic therapy in patients experiencing an acute myocardial infarction. AB - As one of the first hospitals in northwest Arkansas to begin administering thrombolytic therapy to patients with heart attack, or acute myocardial infarction (AMI), Crawford Memorial Hospital (CMH) first administered streptokinase to a patient experiencing an AMI on April 26, 1984. A national standard in lytic therapy was set in 1992 by the National Heart Attack Alert Program Coordinating Committee. The committee set a benchmark of having every appropriate AMI patient receive thrombolytic therapy within 30 minutes of hospital arrival. To monitor quality in lytic therapy administration, CMH began to participate in the National Registry for Myocardial Infarction (NRMI), in January 1995. The first quarter of data revealed a median door-to-drug time (time from arrival at hospital to administration of drug) of 67 minutes. As a quality improvement project, a research experiment was conducted to assess the effect of a case map, also referred to as a clinical pathway, on time to administration of thrombolytic therapy. A case map is a written management plan that provides the ideal sequence and timing of health care staff actions to achieve optimal patient outcomes with minimal variation in care. The researcher developed a case map designed to increase efficiency in delivery of thrombolytic agents. The research was conducted throughout an 18-month period from July 1995 until December 1996. Median time to administration of thrombolytic therapy was reduced from 64 minutes to 25 minutes as a result of case map use (p = 0.028). In this article, research findings are presented regarding the use of case maps in thrombolytic therapy, as well as implications for practice. PMID- 9392267 TI - The RCN unveils the results of its ward leadership project. PMID- 9392268 TI - Lights, camera ... action? PMID- 9392269 TI - Day of reckoning. PMID- 9392270 TI - Digging for labour. PMID- 9392271 TI - My sham trial. PMID- 9392272 TI - Human rights? PMID- 9392273 TI - Free for all. PMID- 9392274 TI - Ward leadership. PMID- 9392275 TI - It's good to talk. Interview by Kate Williams. PMID- 9392276 TI - Clean hands. PMID- 9392277 TI - Standards of nursing care 1987-1997. PMID- 9392279 TI - Trust-wide core care plans. AB - This article discusses the concept and implication of core care plans. The authors also assess the effect of the introduction of core care plans introduction within the Scarborough and North East Yorkshire Healthcare NHS Trust. PMID- 9392278 TI - Evaluation of a nurse-led minor injuries unit. AB - This article, the first of two, reports the evaluation of a minor injuries clinic. The authors describe the background to the service and the methodology and results of the study. Some 20,000 patients attended the clinic during its first two years, at an average cost of 33 pounds per patient. Waiting times were low and 67 per cent of patients were discharged. Independent clinical audit rated 98 per cent of cases as satisfactorily treated. The clinic has generally been accepted by the local medical community and was funded permanently in late 1996. PMID- 9392280 TI - Surviving cancer: a review of the impact and consequences. AB - In this critical review of the literature, the author examines articles assessing the effects on patients of cancer survival. Implications for nursing practice, education and research are also discussed. PMID- 9392281 TI - Comparing asthma and chronic obstructive pulmonary disease (COPD). PMID- 9392282 TI - Plans for a nurse recruitment campaign. PMID- 9392283 TI - Funding shift needs a lesson in clarity. PMID- 9392284 TI - Capital crisis. PMID- 9392285 TI - Don't shoot the tsar. PMID- 9392286 TI - Security guard. PMID- 9392287 TI - People's contract. PMID- 9392288 TI - Bad science. PMID- 9392289 TI - Violence at work. PMID- 9392290 TI - Peaceful means. PMID- 9392292 TI - Learn to surf. PMID- 9392291 TI - Family support. Interview by Dina Leifer. PMID- 9392293 TI - Mental health network: a research programme. AB - This report is based on information supplied by the Network for Psychiatric Nursing Research in Oxford. Each month, we highlight work being carried out in key areas of interest to mental health nurses. This week, we concentrate on a selection of research and views about how psychiatric nurses manage risk. PMID- 9392294 TI - Intravenous sedation for short procedures and investigations. PMID- 9392295 TI - Reducing the risk of complications in i.v. therapy. PMID- 9392296 TI - Minor injuries units: evaluating patients' perceptions. AB - In the second of two articles, the authors report on an evaluation of a minor injuries clinic run by nurse practitioners. The first article was published last week. PMID- 9392297 TI - Violence to nurses: prevalence and risk factors. AB - This article provides a review of current knowledge on the problem of violence to nurses by patients and relatives. In particular, the article discusses the extent of such violence in nursing generally and across different specialties. PMID- 9392298 TI - High-dose/activation-associated tolerance: a mechanism for allograft tolerance. PMID- 9392299 TI - Characterization of pigs transgenic for human decay-accelerating factor. AB - BACKGROUND: To prevent the central role played by complement activation in the hyperacute rejection of pig organs transplanted into primates, pigs transgenic for human decay-accelerating factor (HDAF) have recently been produced. The data presented here extend previous immunohistochemical findings by documenting the immunological characterization and the levels of expression of HDAF in these transgenic pigs. METHODS: Animals from 30 independently derived lines were included in this study. HDAF expression was characterized by immunoprecipitation and epitope mapping. Quantitative analysis was performed by radiometric assays followed by Scatchard analysis and by double-determinant radioimmunoassay. Deposition of iC3b on porcine aortic endothelial cells was determined by radioimmunoassay. DNA slot-blot analysis and densitometric scanning were used to evaluate HDAF transgene copy number. RESULTS: The integrity of HDAF expressed by these transgenic pigs could be demonstrated. HDAF was present in 72% of the organs analyzed, although considerable variation in expression occurred, both between animals and within the same pig. High levels of HDAF on porcine aortic endothelial cells resulted in iC3b deposition at levels as low as that detected on human endothelial cells. Twenty-six organs expressed levels of HDAF greater than those observed in the equivalent human tissue. HDAF expression did not correlate with the number of copies of the transgene incorporated into the porcine genome. CONCLUSIONS: Transgenic pigs, which express levels of functional HDAF even greater than those observed in humans, have successfully been produced. Pigs transgenic for human complement inhibiting molecules could represent a source of organs for future clinical xenotransplantation. PMID- 9392300 TI - Apoptosis after intestinal ischemia-reperfusion injury: a morphological study. AB - BACKGROUND: Apoptosis has been identified after ischemia-reperfusion (IR) injury to the brain, heart, kidney, retina, and the adrenals. Intestinal IR injury causes villous and crypt damage, which has so far been attributed to cellular necrosis. This study was undertaken to investigate the possible role of apoptosis after reperfusion of cold-stored small bowel grafts in syngeneic rats. METHODS: Small intestinal grafts were stored at 4 degrees C for 24 hr in saline (n=6) or in modified University of Wisconsin solution (n=6), followed by reperfusion for 1 hr in syngeneic Lewis rats. Small bowel samples were obtained before storage, after preservation and after 1 hr of reperfusion. They were processed for light and electron microscopy and analyzed for cell death, with particular emphasis on apoptosis. RESULTS: Less than one apoptotic event was seen per 10 crypts in normal and stored bowels. An occasional normal and some denuded villous epithelial cells of stored bowels exhibited apoptosis. After isotransplantation and 1 hr of reperfusion, marked increase in apoptosis was seen in the crypts and denuded villous epithelial cells of both saline- and modified University of Wisconsin-stored bowels. Secondary necrosis was seen in apoptotic cells, as were dark cells. Only a few cells showed signs of primary ischemic necrosis. CONCLUSIONS: Apoptosis occurs after intestinal IR injury. Modulation of its genetic regulatory and biochemical effector machinery might alleviate or even prevent IR injury in small bowel transplanted after similar periods of storage. PMID- 9392301 TI - Up-regulation of oxygen-derived free radicals by interleukin-1 in hepatic ischemia/reperfusion injury. AB - BACKGROUND: Oxygen-derived free radicals (FRs) are critical mediators of ischemia/reperfusion injury. Inflammatory cytokines have been shown to play important roles in tissue injury. To examine the relationship between FRs and interleukin-1 (IL-1) in hepatic ischemia/reperfusion injury, we used interleukin 1 receptor antagonist (IL-1ra) to block endogenous IL-1 production in a rat model of hepatic ischemia/reperfusion. METHODS: Female SD rats were subjected to 30 min of hepatic ischemia followed by reperfusion. The animals were divided into two groups, control group and IL-1ra-treated group, according to the rinse solution. In both groups, FR production, histological changes, and interactions between leukocytes and endothelial cells were analyzed in the course of reperfusion. RESULTS: In the control group, production of FRs increased significantly after 60 min of reperfusion. After 60 and 180 min of reperfusion, histological examination showed atrophy and degeneration of hepatocytes. Hepatic microcirculation demonstrated a marked increase in the number of leukocytes adherent to endothelial cells and of injured cells after reperfusion. In the IL-1ra-treated group, IL-1ra pretreatment markedly reduced FR production after 60 min of reperfusion, the number of leukocytes adherent to endothelial cells, and tissue injury. CONCLUSION: These data clearly show an important role for IL-1 in the induction of FR production, leukocyte adhesion, and tissue injury after hepatic ischemia/reperfusion injury. PMID- 9392302 TI - Pentoxifylline reduces nephrotoxicity associated with cyclosporine in the rat by its rheological properties. AB - BACKGROUND: The goals of this study were to evaluate whether administration of pentoxifylline (POF) reduces the nephrotoxicity associated with cyclosporine (CsA) in the rat, and whether the effect of POF is related to its rheological properties. METHODS: Mean arterial pressure was measured by an intraarterial catheter. Glomerular filtration rate and renal plasma flow were determined by measuring inulin and para-aminohippurate clearances, after double-blind coadministration for 10 days of CsA (25 mg/kg/day) with either vehicle or POF (45 mg/kg every 12 hr). These results were compared with those obtained in control rats. Blood viscosity and erythrocyte deformability were also evaluated after treatment using a cone plate viscometer and a filtration method, respectively. RESULTS: No changes were observed in mean arterial pressure in both groups compared with controls. Glomerular filtration rate was significantly lower in CsA treated rats (0.3+/-0.1 ml/min/100 g) than in control animals (0.6+/-0.1 ml/min/100 g, P<0.02). The coadministration of CsA with POF normalized the glomerular filtration rate (0.6+/-0.1 ml/min/100 g). A parallel decrease in renal plasma flow was observed in CsA-treated rats compared with controls (CsA+vehicle: 1.5+/-0.2 vs. control: 2.2+/-0.1 ml/min/100 g, P<0.02), this effect completely reversed by cotreatment with POF (3.1+/-0.2 ml/min/100 g). Blood viscosity was significantly higher in CsA-treated rats than in the control group (CsA+vehicle: 5.6+/-0.7 vs. control: 5.0+/-0.4 m x Pa x s, P<0.05). This effect was associated with a lower erythrocyte deformability (CsA+vehicle: 1.2+/-0.2 vs. control: 1.5+/ 0.3 ml/min, P<0.05). These rheological abnormalities were normalized by coadministration with POF (blood viscosity: 4.9+/-0.7 m x Pa x s and erythrocyte deformability: 1.9+/-0.4 ml/min, P<0.05). CONCLUSIONS: Our results show that administration of POF prevents the nephrotoxicity associated with CsA. This beneficial effect could be related to its rheological properties. PMID- 9392303 TI - Induction of tolerance toward rat cardiac allografts by treatment with allochimeric class I MHC antigen and FTY720. AB - BACKGROUND: The combination of FTY 720, a novel immunosuppressant, and allochimeric class I MHC proteins bearing donor-type amino acid (aa) epitope substitutions for host-type sequences induces tolerance of Wistar Furth (WF; RT1.Au) heart allografts in ACI (RT1.Aa) recipients. METHODS: Allochimeric alpha(1h)l58-80-RT1.Aa proteins were produced by substituting the allogeneic nucleotide sequence encoding 10 aa residues unique to the alpha1 helical (alpha1h) region of RT1.Al Lewis (Asp58, Arg62, Glu63, Gln65, Lys66, Gly69, Asn70, Asn73, Ser77, and Asn80) for native RT1.Aa residues. The RT1.Au and the RT1.Al haplotypes share four of these aa (Arg62, Glu63, Gln65, and Gly69). A baculovirus/Spodoptera frugiperda insect cell system was used to express the alpha(1h)l58-80-RT1.Aa proteins. RESULTS: The addition of a 3-day oral gavage of 0.05 mg/kg/day FTY720 to a single portal vein injection of 10 microg alpha(1h)l58 80-RT1.Aa protein induced permanent acceptance of WF heart allografts in 16 of 26 ACI recipients (>100 days); the alpha(1h)l58-80-RT1.Aa protein alone only modestly prolonged WF heart survival (13.8+/-0.8 days). The same tolerogenic protocol did not prolong the survival of third-party Brown Norway (RT1.An) heart allografts (14.3+/-2.5 days) compared with FTY720 alone (14.0+/-2.3 days; NS). Tolerant ACI recipients bearing primary WF heart allografts for more than 100 days accepted second WF hearts, but promptly rejected third-party Brown Norway heart grafts (9.3+/-1.5 days). The tolerant state was transferred to irradiated ACI rats (400 rad) with either purified T cells (4-10 x 10[7]) or serum (1-2 ml) from tolerant hosts, and was not broken by daily intraperitoneal injections of interleukin-2 (1000 U/day; 7 days). CONCLUSIONS: The combination of allochimeric protein with FTY720 induces transplantation tolerance, a state that may be associated with the appearance of donor-specific regulatory factors. PMID- 9392304 TI - Expression of an allogeneic MHC DRB transgene, through retroviral transduction of bone marrow, induces specific reduction of alloreactivity. AB - BACKGROUND: Transfer of MHC class II genes, through allogeneic bone marrow (BM) transplantation, induced long-lasting acceptance of renal allografts in miniature swine. To adapt this approach to the clinic, we have now examined whether somatic transfer of allogeneic class II DR genes, into otherwise autologous bone marrow cells (BMC), can provide the matching required for inducing immune tolerance. METHODS: Autologous BMC were transduced ex vivo with recombinant retroviruses for allogeneic DRB followed by BM transplantation. The recipients were then challenged with kidney allografts solely matched to the DRB transgene. RESULTS: Five miniature swine received autologous BMC conditioned with growth factors and transduced with recombinant retrovirus vectors containing allogeneic (n=4) or syngeneic (n=1) class II DRB genes and a drug-resistance marker. Expression of retrovirus-derived products in BM-derived cells was demonstrated by the detection of drug-resistant colony-forming progenitors and the presence of DRB retrovirus transcripts in peripheral cells. Analysis of selective mixed lymphocyte reaction responses to DR or DQ antigens indicated decreased reactivity toward the transduced DR gene product. Among all of the animals receiving fully mismatched kidney allografts, but with DRB matched to the transduced DRB, the one with the highest gene transduction rate showed stable allograft function and essentially normal renal histology for 2.5 years. A control animal, which received a syngeneic DRB gene, rejected its kidney allograft in 120 days after an earlier rejection crisis. CONCLUSIONS: These studies demonstrate that allogeneic MHC gene transfer into BM provides a new strategy for inducing tolerance across MHC barriers. PMID- 9392305 TI - Influence of donor and recipient ages and sex on graft function after pediatric renal transplantation. AB - BACKGROUND: Adult donor grafts adapt to the smaller size of the child recipient by reducing their absolute glomerular filtration rate (GFR) (ml/min). The question arises whether these grafts can increase the absolute GFR when the child recipient grows or whether a child donor graft can better increase its function. The aim of this study was to evaluate the influence of donor and recipient ages and sex on renal function. METHODS: Eighty-five children and adolescents, aged 0.4-20.5 years at transplantation, were monitored annually, by GFR and effective renal plasma flow (ERPF), determined by clearances of inulin and para aminohippuric acid. The patients received 90 grafts from donors aged 3-67 years. Follow-up time was around 5 years. RESULTS: Absolute GFR and ERPF (ml/min) of grafts from donors <20 years of age (all cadaveric donor grafts) increased during follow-up, resulting in a constant relative GFR and ERPF (ml/min/1.73 m2), whereas absolute GFR and ERPF of grafts from donors >20 years of age remained constant during follow-up, resulting in a significant decrease in relative values. Relative GFR and ERPF fell during follow-up in young recipients (<12 years of age), but remained constant in older recipients (>12 years). Donor and recipient sex did not influence renal function. CONCLUSIONS: Child donor grafts seem better able to increase their function with the growth of the child recipient than adult grafts. However, the limited access to pediatric grafts and the fact that pediatric cadaveric grafts might involve technical problems in connection with grafting restrict their use. PMID- 9392306 TI - Efficacy of OKT3 as primary therapy for histologically confirmed acute renal allograft rejection. AB - BACKGROUND: OKT3 is often used as primary treatment for acute renal allograft rejection. In a retrospective study, we sought to determine the efficacy of OKT3 as a first-line agent in reversing histologically confirmed acute renal allograft rejection. METHODS: Patients with mild to moderate, moderate, or severe acute cellular and acute vascular rejection who had not received any other anti rejection treatment were included in this analysis. A total of 88 patients, who received OKT3 between 1987 and 1995, fulfilled these criteria. RESULT: Seventy of these patients were renal transplant recipients, and 18 were combined kidney and pancreas transplant recipients. The median time to the diagnosis of rejection from transplantation was 32 days (range, 6 days to 13 years). On histology, 6 were graded as mild to moderate, 36 as moderate, 29 as moderate to severe, and 17 as severe rejection. The mean baseline serum creatinine was 1.62 mg/dl (range, 0.7-10.1 mg/dl), and the mean serum creatinine at the time of diagnosis of rejection was 2.60 mg/dl (range, 1.4-12.7 mg/dl) (P=<0.0001). The mean duration of OKT3 treatment was 11.2 days (range, 8-18 days). The mean serum creatinine at the end of OKT3 treatment was 1.73 mg/dl (range, 0.6-5.0 mg/dl; P=0.24 compared with baseline serum creatinine). Rejection was reversed in 86 (98%) patients. Graft survival at 1 year after OKT3 therapy was 87.5% (77 of 88). At a mean follow-up of 38 months, 8 patients had died and 26 grafts were lost. The mean serum creatinine level in the 64 patients with a functioning graft was 1.76 mg/dl (range, 0.8-4.0 mg/dl) at the last follow-up. CONCLUSION: OKT3 when utilized as first-line therapy reversed 98% of the acute rejection episodes, with a 1-year post-OKT3 graft survival of 87.5%. PMID- 9392307 TI - Persistent long-term changes in lymphocyte subsets induced by polyclonal antibodies. AB - BACKGROUND: Clinicians are well aware of the short-term effects of immunosuppression by mono- or polyclonal antibodies. Little is known about long term changes induced by these therapies. METHODS: Forty-three renal allograft recipients were selected according to their initial postoperative immunosuppression: (1) BI group=basic immunosuppression with steroids and cyclosporine, n=16; (2) ATG group=basic immunosuppression plus polyclonal antibody antithymocyte globulin (ATG), n=11; and (3) OKT3 group=basic immunosuppression plus monoclonal antibody OKT3, n=16 patients. At intervals of 6 months, the following parameters were measured prospectively: lymphocyte surface antigens (HLA-DR, CD3, CD4, CD8, CD16, CD19, CD56, and CD57); serum and urine neopterin; serum amyloid A; and indirect and direct tests for herpes viruses. RESULTS: The mean period of observation was 58.4 months. The most significant differences between the groups occurred for CD4+ and CD8+ T cells. The ratios of CD4+ to CD8+ cells (n=278 measurements) were significantly and persistently lower in the ATG group (P<0.001, Brown-Mood test). Five years after transplantation, the ATG group had a CD4+ to CD8+ cell ratio of x=0.6 versus x=1.7 in the OKT3 group and x=2.0 in the BI group. This inversion was due to a persistent depletion of the CD4+ cells and an increased regeneration of the CD8+ cells, in particular of the CD8+brightCD57+ subpopulation. Extent and duration of CD4+ depletion correlated with the cumulative ATG dose (r=0.7, P<0.05, Spearman rank correlation test). CONCLUSION: Therapy with polyclonal antibody ATG induces dose-dependent long-term changes in T-cell lymphocyte subsets, which persist over a period of years. PMID- 9392308 TI - Early signs and risk factors for the increased incidence of Epstein-Barr virus related posttransplant lymphoproliferative diseases in pediatric liver transplant recipients treated with tacrolimus. AB - BACKGROUND: Posttransplant lymphoproliferative disease (PTLD) is a life threatening condition the incidence of which in pediatric solid organ transplantation may be related to the immunosuppressive load. It has been suggested that tacrolimus, a new and potent immunosuppressor, causes an increased incidence of this syndrome. METHODS: The incidence, early signs, and risk factors for lymphoproliferative disease were reviewed in a cohort of 89 pediatric liver transplant recipients treated with tacrolimus. RESULTS: Eighteen patients (20%) developed a PTLD-16 concomitant to a primary Epstein-Barr virus (EBV) infection and 2 with previous immunity against EBV. Three additional patients had preliminary signs of PTLD concomitant to primary EBV infection, but did not develop individualized lymphoid masses. Six patients died (6.7% of all tacrolimus treated patients). Mean tacrolimus blood level during the 3 months preceding EBV infection reached 11.8+/-1.8 ng/ml in PTLD patients versus 9.4+/-3.4 ng/ml in non PTLD patients (0.0550% above the upper limit of the reference range at a median of 1 day before the equivalent change in alanine transaminase in association with allograft rejection in the combined groups (95% confidence interval=1 to 2 days) but was lower on the day of diagnosis of rejection in the reporting group (P=0.02). This is compatible with the earlier diagnosis of rejection in the reporting group. CONCLUSIONS: We conclude that the monitoring of GST may improve patient care, reducing both mortality and morbidity. PMID- 9392311 TI - Expansion of CD4+CD7- T cells, a memory subset with preferential interleukin-4 production, after bone marrow transplantation. AB - BACKGROUND: Inadequate reconstitution of CD4+ lymphocyte and interleukin (IL)-2 production defect are observed after bone marrow or peripheral blood stem cell transplantation (SCT). METHODS: We studied immune reconstitution after SCT in 33 consecutive patients who received allogeneic SCT (17 patients) or autologous SCT (16 patients). The aims were to assess the regeneration of the CD4+ T-cell subset with regard to helper cell differentiation. CD4+ T-cell subset regeneration and expansion of the CD4+CD7- subset were studied by immunofluorescence analysis. CD4+CD7- cell cytokine secretion was analyzed after cell sorting and costimulation of the CD3 and CD28 pathways, in enzyme-linked immunosorbent assay and reverse transcription-polymerase chain reaction assays. RESULTS: We report a relative expansion of the CD4+CD7- subset within CD4+ T cells, detected as early as 1 month after bone marrow transplantation and decreasing after day 60. CD4+CD7 T cells preferentially expressed CD45RO and activation markers such as CD57, CD25, and HLA-DR. No relationship was observed between the CD4+CD7- expansion and transplant-related complications. We observed no significant IL-2 production in supernatants from sorted CD4+CD7- T cells, whereas IL-4 levels were comparable to those produced by cells from normal individuals. Autologous CD4+CD7+ cells showed little, if any, IL-4 production, and IL-2 production was lower than that by normal CD4+CD7+ T cells. Reverse transcription-polymerase chain reaction assays showed similar amounts of interferon-gamma transcripts in the two subsets; tumor necrosis factor-alpha, IL-4, and IL-10 transcripts were detected in CD4+CD7- T cells but not in their CD4+CD7+ counterparts. CONCLUSIONS: These data confirm the IL-2 production defect after bone marrow transplantation and suggest that the CD4+CD7- T-cell subset might be preferentially involved in the enhanced production of IL-4 and low production of IL-2. These data show that the early immune reconstitution in CD4+ T cells after SCT preferentially involves memory T cells with a Th0/Th2 differentiation that might participate in the T-helper cell defect. PMID- 9392312 TI - Immunosuppression by D-isomers of HLA class I heavy chain (amino acid 75 to 84) derived peptides is independent of binding to HSC70. AB - BACKGROUND: Peptides derived from the class I heavy chain were shown to modulate immune responses in vitro and in vivo. A peptide derived from HLA-B2702 (2702.75 84) inhibited differentiation of cytotoxic T cells as well as T cell and natural killer cell-mediated cytotoxicity in vitro. Peptide-mediated immunomodulation seemed to be independent of the MHC proteins expressed by responder and stimulator cells. In vivo studies in rodents demonstrated prolongation of heart and skin allograft survival after peptide therapy. Here, the correlation between the peptide's biological activity and its amino acid sequence was analyzed using peptides derived from amino acid 75-84 of several mouse, rat, and human MHC class I proteins as well as peptides with single amino acid substitutions in the 2702.75-84 sequence. METHODS: Peptides consisting of both L- and D-amino acids were tested for inhibition of murine and human T cell-mediated and lymphokine activated killer cell-mediated cytotoxicity, binding to hsc70, and prolongation of heart allograft survival in vivo. RESULTS: Replacement of glutamic acid residue (E) at position 75 with valine (V) resulted in a peptide [2702.75 84(E>V)] with increased in vitro and in vivo activity but unchanged affinity for hsc70. Surprisingly, both L- and D-isomers of 2702.75-84 and 2702.75-84(E>V) inhibited cytotoxic cells in vitro and prolonged heart allograft survival in vivo. However, as expected, the peptides consisting of D-amino acids did not bind to hsc70. CONCLUSION: Assuming that both D- and L-isomers modulate immune responses by similar mechanisms, these results suggest that the peptides' effect is independent of binding to hsc70. PMID- 9392313 TI - An increase in peripheral blood progenitor cells in primates by coadministration of recombinant human interleukin 6 and recombinant human granulocyte colony stimulating factor. AB - BACKGROUND: We recently demonstrated that coadministration of recombinant human interleukin 6 (rhIL-6) and recombinant human granulocyte colony-stimulating factor (rhG-CSF) in mice synergistically increases peripheral blood stem cells (PBSC), which can rescue lethally irradiated recipient mice. However, there is little information about the effect of coadministration of rhIL-6 and rhG-CSF on PBSC in a primate system. METHODS: In cynomolgus monkeys, rhG-CSF (5 microg/kg day) alone was administered for 5 days (first cycle). After the wash-out period, rhIL-6 (0, 10, or 20 microg/kg/day) and rhG-CSF were coadministered for 5 days (second cycle). RESULTS: Total peripheral colony-forming cells levels were increased earlier by coadministration of rhIL-6 and rhG-CSF than by the administration of rhG-CSF alone. The maximum level in the coadministration cycle was obtained on day 5, and a high level was obtained for a further 3 days after cessation. The maximum number of peripheral total colony-forming cells in the coadministration cycle was a mean of 2.12-fold (range 1.38 to 3.35) higher than that in the rhG-CSF alone cycle. Coadministration also increased the peripheral mixed colony-forming units by a mean of 3.62-fold (range 1.02 to 5.52). Interestingly, monkeys that showed a low response to the administration of rhG CSF alone also had a higher response to the coadministration. No significant difference was observed between the two cycles of administration of rhG-CSF alone. CONCLUSIONS: These findings suggest that coadministration of rhIL-6 and rhG-CSF may be useful for clinical PBSC collection. PMID- 9392314 TI - Impact of HLA compatibility on survival of kidney transplants from unrelated live donors. AB - BACKGROUND: Kidney transplants from unrelated live donors have been reported to perform exceedingly well despite poor HLA compatibility. These results were extrapolated to indicate that HLA matching in cadaver kidney transplantation is superfluous provided the ischemic preservation time is kept very short. METHODS: The influence of HLA matching on the outcome of 2281 transplants from unrelated live donors from 1986 to 1995 was analyzed at 198 transplant centers. RESULTS: The 5-year transplant success rate was significantly associated with the number of HLA-A, -B, and -DR mismatches (P<0.0001). There was a significant impact on graft survival of mismatches at the class I HLA-A, and -B loci (P=0.001) as well as at the class II HLA-DR locus (P=0.005). In transplants from both spouses and nonspouses, cases with no HLA-DR mismatch were found more often than would be expected if transplant donors were selected by chance, which indicates that efforts of prospective HLA matching had been made. CONCLUSIONS: The outcome of kidney transplants from unrelated live donors is strongly influenced by HLA compatibility. Short exposure of donor kidneys to ischemia does not eliminate the influence of HLA matching on graft survival. PMID- 9392315 TI - Effect of troglitazone on blood insulin levels after pancreas transplantation with systemic venous drainage in rats. AB - BACKGROUND: Troglitazone is a new oral antidiabetic agent and has been reported to reduce insulin resistance and improve peripheral hyperinsulinemia in patients with noninsulin-dependent diabetes mellitus. To examine the effect of troglitazone on insulin regulation after pancreas transplantation with systemic venous drainage, we measured peripheral glucose and insulin levels and performed an intravenous glucose tolerance test. METHODS: We divided the rats into four groups: diabetic rats with a pancreas graft and administration of troglitazone at 40 mg/day orally (group P+T, n=4), rats with a pancreas graft only (group P, n=4), age-matched normal rats (group N, n=5), and diabetic rats (group DM, n=4). RESULTS: Fasting insulin levels in group P were relatively higher than those in group N, whereas the values in group P+T were normalized. In the intravenous glucose tolerance test, troglitazone clearly regulates sigma immunoreactive insulin levels of pancreas transplanted rats (P vs. P+T: 244+/-23 vs. 145+/-14 microU/ml, P<0.05). CONCLUSION: Hyperinsulinemia induced by systemic venous drainage, which may progress atherosclerosis, can be controlled with troglitazone treatment. PMID- 9392316 TI - Common bile duct stenosis caused by chronic pancreatitis after liver transplantation for alcoholic cirrhosis. AB - BACKGROUND: The prevalence of chronic pancreatitis in patients with alcoholic cirrhosis ranges from 7% to 11% and is not considered a contraindication for liver transplantation. METHODS: Among 59 liver transplant recipients grafted for alcoholic cirrhosis, we report two observations of common bile duct stenosis due to chronic pancreatitis. RESULTS: In both cases, pretransplant work-up disclosed no clinical or radiological evidence of chronic pancreatitis. The diagnosis of common bile duct stricture was made 6 and 60 months after liver transplantation. One patient was reoperated upon, and his choledochocholedochostomy was converted into a Rouxen-Y choledochojejunostomy. The second patient experienced metastatic laryngeal carcinoma and died before reoperation. CONCLUSIONS: These observations suggest that common bile duct stricture caused by chronic pancreatitis may occur after liver transplantation for alcoholic cirrhosis, even after a long-standing history of abstinence. PMID- 9392317 TI - Persistent hypersplenism early after liver transplant: the role of splenectomy. AB - BACKGROUND: Transient thrombocytopenia is common after liver transplantation, but persisting thrombocytopenia worsens the prognosis after transplant. METHODS: Two patients underwent splenectomy for persistent thrombocytopenia early after liver transplantation. The first patient had a platelet count of 17,000/mm3 on postoperative day (POD) 6; her hemoglobin and white blood cell counts were normal. Work-ups including bone marrow aspiration, Coombs test, and antiplatelet antibody test were negative. On POD 9, she had abdominal bleeding with a platelet count of 17,000/mm3 despite repeated platelet transfusions, and splenectomy was done. The second patient had a platelet count of 3000/mm3 on POD 14, white blood cell was 1600/mm3, and hemoglobin was 7.7 g/dl. Bone marrow biopsy revealed hypercellular marrow. Because his platelet count remained at 2000/mm3 despite empiric treatment with intravenous immune globulin and methylprednisolone, splenectomy was performed. RESULTS: The first patient's platelet count rose to 155,000/mm3 by POD 8. The second patient's platelet count reached 210,000/mm3 on POD 5. Neither patient has had an episode of thrombocytopenia at 36 and 32 months after splenectomy. CONCLUSIONS: Splenectomy can be used after liver transplantation for severe, persistent thrombocytopenic states that cannot be attributed to sepsis, intravascular coagulation, immunological causes, or drug effects. PMID- 9392318 TI - Adenovirus hepatitis in the adult allograft liver. AB - BACKGROUND: Adenovirus hepatitis in the allograft liver is an uncommon condition hitherto recognized only in pediatric patients. We describe two adult cases. METHODS: Clinical information was obtained by reviewing the medical records. The diagnosis of adenoviral infection was made by immunohistochemistry or culture. RESULTS: Both patients had received recent antirejection treatment and presented with fever, hepatic dysfunction, and progressive leukopenia. One patient had some viral inclusions resembling those described in herpes simplex infections. Adenovirus was cultured from the liver in both cases and from the lung in one case. Both patients were treated by decreasing the immunosuppression and intravenous acyclovir, but died. CONCLUSIONS: Adenovirus infection should be considered when evaluating adult liver transplant patients with necrotizing lesions or microabscess formation at allograft biopsy. A review of the literature shows that most previously reported infections have led to graft loss or death, but occasional remissions of disease are also on record. PMID- 9392319 TI - Calcineurin activity in children with renal transplants receiving cyclosporine. AB - BACKGROUND: The major immunosuppressive effect of cyclosporine is through the inhibition of calcineurin, an enzyme important in the activation of T lymphocytes. In children, neither calcineurin activity nor its inhibition by cyclosporine (CsA) has been investigated. METHODS: Calcineurin activity, was measured in stable pediatric renal transplant patients, with healthy children used as controls. Whole blood CsA concentrations were measured by monoclonal radioimmunoassay. Simultaneous calcineurin and CsA levels were measured before and 1, 2, 3.5, 5, and 12 hr after their routine morning CsA dose. RESULTS: Calcineurin activity was approximately 50% inhibited at trough blood concentrations (148 microg/L); moreover, inhibition increased as CsA concentrations rose and declined as concentrations fell. Maximum calcineurin inhibition was about 70% at concentrations of about 431 microg/L. Linear regression analysis revealed a significant correlation between mean CsA blood concentration and the mean degree of inhibition of calcineurin activity (P=0.005, one-tailed). CONCLUSION: We conclude that inhibition of calcineurin activity by CsA in pediatric renal transplant recipients correlates with CsA blood concentrations. PMID- 9392320 TI - Focal acute tubular necrosis in a renal allograft. AB - Nuclear imaging is used to evaluate renal allografts demonstrating delayed function after transplantation. Interpretation of the nuclear scan in the context of clinical data, provides helpful information in the management of the transplant recipient. The better quality of images obtained with technetium-99m mercaptoacetyltriglycine (Tc-99m MAG3) has made it the radiotracer of choice compared to technetium-99m diethylenetriamine pentaacetic acid (Tc-99m DTPA) for imaging of the renal allograft. Tc-99m MAG3 is cleared from the kidney by tubular secretion, whereas Tc-99m DTPA is cleared by glomerular filtration. In this report, we discuss a unique abnormality found on nuclear imaging of a renal allograft. Utilizing our understanding of the characteristic handling of various radiotracers by the kidney, we were able to demonstrate that the renal scan was consistent with an area of focal acute tubular necrosis in the newly transplanted kidney. PMID- 9392321 TI - Recurrent immunoglobulin A nephropathy after renal transplantation: a significant contributor to graft loss. AB - BACKGROUND: Although most transplanted patients with underlying IgA nephropathy (IgAN) develop histological recurrence, its clinical relevance is considered low. METHODS: We performed a single-center analysis of 61 renal transplant patients with IgAN. RESULTS: Forty-four percent of the patients showed a stable graft function. Progressive graft dysfunction apparently due to recurrent IgAN occurred in 23% of the patients (16% required dialysis). Five patients were retransplanted, and three again developed dialysis-dependent renal failure apparently due to recurrent IgAN. In 20% of the patients, chronic transplant dysfunction was due to other reasons, whereas no reason was identified in 13% of the patients. Neither findings before transplantation, the ACE genotype, the type of immunosuppression, nor the course after transplantation predicted transplant dysfunction due to recurrent IgAN. Follow-up after transplantation was longer in the group with dysfunction due to recurrent disease than in the group with dysfunction due to chronic rejection or in the stable group. CONCLUSION: Recurrent IgAN is a clinically relevant problem in renal transplant patients. Its importance may have been underestimated in the past due to inadequate lengths of follow-up. PMID- 9392322 TI - Differential down-regulation of CD28 by B7-1 and B7-2 engagement. AB - Physiologically relevant full activation of T cells requires signal transduction through the T cell receptor and additional costimulatory cell surface molecules. Best understood of these costimulatory interactions are those between CD28 and its ligands B7-1 (CD80) and B7-2 (CD86). While B7-1 and B7-2 bind the same receptors (CD28 and CTLA-4), they share only 25% sequence homology, are expressed at different times during immune responses, and in some systems have been shown to differentially affect T cell cytokine expression. Although CD28 is an activation antigen, its expression is down-regulated after engagement by B7-1. Here we show that B7-2 engagement is considerably less effective at down regulating CD28, which indicates a differential effect of these two CD28 ligands on activated T cells. PMID- 9392323 TI - Comment on "Delayed correction of portal hypertension after portal vein conduit arterialization in liver transplantation" by Neelamekam et al. PMID- 9392324 TI - Increased benefit from shorter screening mammography intervals for women ages 40 49 years. PMID- 9392325 TI - An interdisciplinary approach to avoid the overtreatment of patients with central nervous system lesions. PMID- 9392326 TI - Cytologic detection of esophageal squamous cell carcinoma and precursor lesions using balloon and sponge samplers in asymptomatic adults in Linxian, China. AB - BACKGROUND: The principal reason for the poor prognosis of esophageal carcinoma is that most tumors are asymptomatic and go undetected until they are unresectable. Previous studies have shown that cytologic screening of asymptomatic high risk individuals can detect curable esophageal carcinomas and precursor lesions, but the sensitivity of such screening is not well documented. The current study evaluated the sensitivity and specificity of currently available balloon and sponge cytologic samplers for detecting biopsy-proven squamous dysplasia and carcinoma in asymptomatic individuals from a high risk population in Linxian, China. METHODS: Asymptomatic adults were examined with both balloon and sponge samplers, in random order, followed by endoscopy with mucosal iodine staining and biopsy of all unstained lesions. The cytology slides were interpreted using the criteria of the Bethesda System. The balloon and sponge cytologic diagnoses (test) were compared with the biopsy diagnosis (truth) in each patient to estimate the sensitivity and specificity of each sampler. RESULTS: Of the 439 patients with adequate biopsies, 123 (28%) had histologic squamous dysplasia and 16 (4%) had an invasive squamous carcinoma. The sensitivities/specificities of the balloon and sponge were 44%/99% and 18%/100%, respectively, for detecting biopsy-proven squamous cell carcinoma, and 47%/81% and 24%/92%, respectively, for identifying squamous dysplasia or carcinoma. CONCLUSIONS: In this study, the balloon sampler was more sensitive than the sponge sampler for detecting esophageal squamous disease, but both techniques were less than optimal. Improved samplers and/or cytologic criteria should increase the sensitivities observed in this baseline study. PMID- 9392327 TI - Does dual infection by hepatitis B and C viruses play an important role in the pathogenesis of hepatocellular carcinoma in Japan? AB - BACKGROUND: There are contradictory data concerning the synergistic effect of hepatitis B virus (HBV) and hepatitis C virus (HCV) infection on the progression from chronic hepatitis to hepatocellular carcinoma (HCC). METHODS: To clarify the role of coinfection with HBV and HCV in the progression and pathogenesis of HCC, viral and clinicopathologic features were studied in 368 consecutive HCC patients at the University of Tokyo from 1991-1995. RESULTS: Approximately 83% of patients (305 patients) were seropositive for the HCV antibody ("C-viral") and approximately 10% (37 patients) were positive for the hepatitis B surface antigen ("B-viral"). Positivity for both (dual infection) was found in only 2% of patients, and negativity for both in 5%. The incidence of dual infection in HCC patients was Similar to that in 549 patients with chronic hepatitis (1%) and 119 patients with cirrhosis (1%). Of the six HCC patients with dual infection, five patients were positive for the HBV early antigen and HBV DNA was less than measurable, whereas HCV RNA was detected and ranged from 10(3)-10(6) copies/50 microL of serum by competitive reverse transcriptase-polymerase chain reaction, and the clinical features resembled those of "C-viral" HCC. The remaining patient was early antigen positive and had HBV DNA by slot blot analysis, but the serum HCV RNA level was less than measurable. These data indicate that mutually exclusive viral replication occurred in patients with persistent coinfection. To further clarify further the possible involvement of HBV infection in "C-viral" HCC, HBV core antibody (HBcAb) was tested in 192 patients and was found to be positive in 111 and negative in 81. The serum HCV RNA level and clinicopathologic features (such as age and the severity of liver disease) were similar among the "C-viral" HCC patients irrespective of the presence or absence of HBcAb. CONCLUSIONS: Based on these results, coinfection was found to be much less prevalent than generally is claimed, and even in a few HCC patients with the coinfection the mutually exclusive viral replication was noted, suggesting that coinfection plays little if any role in the development of HCC. PMID- 9392328 TI - Thrombin receptor activation results in calcium signaling and protein kinase C dependent stimulation of DNA synthesis in HEp-2g laryngeal carcinoma cells. AB - BACKGROUND: Recently, the expression of the "tethered ligand" thrombin receptor in carcinosarcoma and melanoma cells has been shown. However, the role of the thrombin receptor in tumor cell metabolism still is undefined. METHODS: In this article, the "tethered ligand" thrombin receptor was identified on human epidermoid carcinoma cells (HEp-2g cell line) by using immunofluorescence studies with a monoclonal antithrombin receptor antibody and radioligand binding. Furthermore, the effects of alpha-thrombin and thrombin receptor activating peptides (TRAP)-6 on calcium mobilization, protein kinase C (PKC) translocation, and DNA synthesis were estimated. RESULTS: Pharmacologic characterization using [3H]TRAP-6 as a radioligand demonstrated a single class of high affinity binding sites (dissociation constant [KD] = 7.2 +/- 2.2 x 10(-7) M) and a binding capacity of 27 +/- 3.4 fmol/mg protein. The function of these binding sites was demonstrated by alpha-thrombin- and TRAP-6-induced mobilization of free intracellular calcium, and translocation of PKC from cytosol to cell membrane. Moreover, alpha-thrombin and TRAP-6 induced an increase in [3H]thymidine incorporation in HEp-2g cells that could be blocked by the PKC inhibitor bisindolylmaleimide. CONCLUSIONS: To the authors' knowledge, the results of this study demonstrate for the first time functional thrombin receptors in epidermoid carcinoma cells. The thrombin receptor appears to be involved in growth regulation in HEp-2g cells by a PKC-dependent mechanism. PMID- 9392329 TI - Stereotactic radiosurgery for patients with nonsmall cell lung carcinoma metastatic to the brain. AB - BACKGROUND: A retrospective study of patients undergoing stereotactic radiosurgery for one to four brain metastases from nonsmall lung cell carcinoma (NSCLC) was performed to document outcomes and risks. METHODS: Seventy-seven patients underwent radiosurgery during a 7-year interval; 71 also underwent whole brain radiation therapy. Univariate and multivariate analyses were used to determine significant prognostic factors affecting survival. RESULTS: The overall median survival was 10 months after radiosurgery, and 15 months from the diagnosis of brain metastases. Five factors significantly affected survival: extent of systemic disease, presence of a neurologic deficit, size of the intracranial tumor, initial imaging appearance of intratumoral necrosis, and initial resection of the primary tumor of the chest. Median survival time was 26 months in a subgroup of patients with no extracranial metastases, no neurologic deficits, and a small tumor without necrosis. The authors evaluated 91 tumors with imaging. Local tumor control was achieved in 77 lesions (85%) and tumoral radiation necrosis developed in 4 lesions (4.4%). Nineteen new metastatic tumors developed during the observation interval. CONCLUSIONS: Stereotactic radiosurgery for NSCLC brain metastases is effective and is associated with few complications. The early detection of brain metastases and treatment with radiosurgery combined with radiation therapy provide the opportunity for extended high quality survival. PMID- 9392330 TI - Isolated limb reperfusion with tumor necrosis factor and melphalan in patients with extremity melanoma after failure of isolated limb perfusion with chemotherapeutics. AB - BACKGROUND: This retrospective study evaluated the benefit of using tumor necrosis factor (TNF) and melphalan administered via an isolated limb perfusion (ILP) in a series of patients with metastatic melanoma who failed initial ILP with chemotherapeutics. METHODS: Seventeen patients with extremity melanoma who underwent prior ILP with conventional chemotherapeutics (10 with melphalan; 4 with platinum; 2 with platinum, dacarbazine, thiotepa, actinomycin D, and nitrogen mustard; and 1 with thiotepa, actinomycin D, and nitrogen mustard) and had local recurrences were treated with a 90-minute isolated hyperthermic limb reperfusion with melphalan (10 mg/L limb volume) plus TNF (2-6 mg). Five prior ILPs were adjuvant and 12 were therapeutic. RESULTS: Reperfusion was associated with an overall 94% response rate and a 65% complete response (CR) rate. Of the patients who failed an initial ILP with melphalan alone the overall response rate was 90% after the reperfusion with TNF and melphalan. In patients who failed an initial ILP with agents other than melphalan the CR rate was 100% after ILP with TNF and melphalan. TNF/melphalan isolated limb reperfusion was found to be more effective in terms of CR after initial ILP regimens that did not utilize melphalan (100% CR after nonmelphalan ILP vs. 50% CR after melphalan ILP [P = 0.04]). Regional toxicity was comprised of mild skin blistering and peeling in 47% of patients. One patient developed Grade 3 (based on National Cancer Institute Common Toxicity Criteria) skin necrosis, and one developed Grade 5 muscle and nerve toxicity, requiring an amputation. CONCLUSIONS: Isolated limb reperfusion with TNF and melphalan can be performed safely with response rates similar to those of other trials of single perfusions. Repeat ILP using TNF and melphalan in patients with melanoma who have failed prior ILP with chemotherapeutics is justified. The utility of TNF (vs. melphalan alone) will be defined in ongoing Phase III trials. PMID- 9392331 TI - The Gothenburg breast screening trial: first results on mortality, incidence, and mode of detection for women ages 39-49 years at randomization. AB - BACKGROUND: The effect of mammography screening on breast carcinoma mortality in women ages < 50 years remains unclear. METHODS: A randomized trial of invitation to breast carcinoma screening with mammography was performed in the city of Gothenburg, Sweden. The purpose was to estimate the effect of mammographic screening on breast carcinoma mortality in women ages < 50 years. Randomization was initially by day-of-birth cluster (18% of subjects), and subsequently by individual (82% of subjects). Between September 1983 and April 1984, 11,724 women ages 39-49 years were randomized to the study group. This group was invited to mammographic screening every 18 months. Two-view mammography was used at each screen unless the density of the breast at the previous screen indicated that single view was adequate. Fourteen thousand two hundred and seventeen women in the same age range were randomized to a control group that was not invited to undergo screening until the fifth screen of the study group (between 6 and 7 years after randomization). Women with breast carcinoma diagnosed up to the time immediately after the first screen of the control group were followed for death from breast carcinoma until the end of December 1994. RESULTS: A 45% reduction in mortality from breast carcinoma was observed in the study group compared with the control group (relative risk [RR] = 0.55, P = 0.035, 95% confidence interval [CI], 0.31-0.96). A conservative estimate based on removal of the tumors detected at the first screen of the control group gave a mortality reduction of 44% (RR = 0.56, P = 0.046, 95% CI, 0.31-0.99). In both cases, the effect was statistically significant. CONCLUSIONS: Mammographic screening can reduce mortality from breast carcinoma in women ages < 50 years. The mortality reduction can be substantial if high quality mammography is used and an 18-month interscreening interval is strictly adhered to. PMID- 9392332 TI - Analysis of bcl-2 in sporadic breast carcinoma. AB - BACKGROUND: The bcl-2 gene encodes a protein that blocks apoptosis and might help to promote tumor development. It is expressed in a high percentage of breast tumors and is associated with good prognostic features. However, the mechanisms that regulate bcl-2 expression in breast carcinoma are unknown. Moreover, immunohistochemical detection of bcl-2 is related inversely to p53 expression. This notwithstanding, the immunohistochemical detection of p53 does not always correlate with the detection of p53 gene mutations. The authors studied the molecular organization of bcl-2 as well as the methylation status of its CpG island and analyzed the correlation between bcl-2 expression and p53 gene mutations. METHODS: The molecular organization of the bcl-2 gene and the methylation pattern of its CpG island were analyzed by Southern blot analysis. In addition, immunohistochemical analysis of bcl-2 and p53 protein expression was performed. Finally, the presence of mutations at exons 5-9 of the p53 gene were analyzed by polymerase chain reaction and single-strand conformation polymorphism. RESULTS: No molecular abnormality was found at the bcl-2 locus in cases of sporadic breast carcinoma. Moreover, loss of heterozygosity analysis failed to detect any allelic loss in the study cases. It also was found that the bcl-2 CpG island was demethylated in all cases. These results point to a lack of correlation between bcl-2 protein expression and the presence of p53 gene mutations. CONCLUSIONS: The level of bcl-2 expression in breast carcinoma is not associated with any somatic abnormality or epigenetic change at the bcl-2 locus. Conversely, although bcl-2 expression is related inversely to p53 protein expression, the analysis of p53 mutations (limited to exons 5-9) failed to demonstrate any relationship between p53 mutations and bcl-2 protein expression. PMID- 9392333 TI - Cathepsin D and chromogranin A as predictors of long term disease specific survival after radical prostatectomy for localized carcinoma of the prostate. AB - BACKGROUND: The accumulation of chromogranin A (Chr A) and cathepsin D (Cath D) gene products may be important in prostate carcinoma progression. This study assessed whether the levels of immunoreactivity for Chr A and Cath D are better predictors of disease specific survival than conventional pathologic parameters of the primary tumor such as Gleason score, capsular penetration, seminal vesicle invasion, and percent tumor in the specimen for patients with clinically localized prostate carcinoma managed by radical prostatectomy. METHODS: Seventy one patients with modified Jewett clinical stages A1 to B2 adenocarcinoma of the prostate underwent a radical prostatectomy after a negative metastatic workup. No neoadjuvant or adjuvant treatments were given and all disease recurrences and causes of death were recorded. Analysis of prostatectomy specimens was undertaken to determine the conventional pathologic parameters of the primary tumor and Chr A and Cath D immunohistochemical staining. Univariate and multivariate analyses were performed to determine the independent contributions of Chr A and Cath D in predicting survival. RESULTS: On univariate analysis Chr A was the only variable that reached statistical significance for disease specific survival (P = 0.035). Cath D nearly reached significance with a P value of 0.079 for disease specific survival. On multivariate analysis, the only independent factor predicting disease specific survival was the Chr A staining score (P < 0.05). In patients with unequivocal foci of Chr A immunoreactivity, the 14-year disease specific survival was 50% compared with 68% for patients lacking such foci. CONCLUSIONS: The level of Chr A immunohistochemical staining is a strong predictor of disease specific survival and is superior to standard pathologic prognostic factors. Such findings lay the groundwork for future prospective study of the utility of such markers on biopsy specimens to predict patient outcome. PMID- 9392334 TI - Testicular "tumor" of the adrenogenital syndrome: a case report of an unusual association with myelolipoma and seminoma in cryptorchidism. AB - BACKGROUND: Males with congenital adrenal hyperplasia may develop bilateral testicular masses in early adult life. These are not malignant and generally regress with corticosteroid therapy. The authors report a case occurring in a 44 year-old man with associated seminoma and myelolipoma in an undescended testis. METHODS: The testicular tumors were analyzed by histologic, flow cytometric, and ultrastructural techniques. RESULTS: The tumors in both testes were comprised of polygonal cells with abundant granular eosinophilic cytoplasm, occasionally with brown (lipochrome) pigment and round nuclei of various sizes with prominent nucleoli. These cells were grouped into nodules by dense and sometimes thick fibrous trabeculae in the right testis. The areas corresponding to the fibrous trabeculae in the left (intraabdominal) testis were replaced by mixture of hematopoietic (myeloid) and fatty tissue in various proportions characteristic of myelolipoma. The left testis also had a well demarcated tumor that was diagnostic of seminoma. Electron microscopy demonstrated abundant smooth endoplasmic reticulum, a moderate number of mitochondria with tubulovesicular cristae, lipid droplets, and lipofuscin granules in the polygonal cells. No Reinke's crystals were observed. The patient received corticosteroids for his adrenocorticoid deficiency and also underwent external beam irradiation to the retroperitoneum for seminoma. CONCLUSIONS: This case illustrates an unusual presentation of a testicular tumor in a patient with the adrenogenital syndrome as well as with myelolipoma and seminoma in a cryptorchid testis. The possibility of an associated neoplasm that could be potentially fatal should be considered whenever a testicular tumor of the adrenogenital syndrome continues to grow despite adequate hormonal treatment. PMID- 9392336 TI - Immunohistochemical analysis of progesterone receptor and Ki-67 labeling index in astrocytic tumors. AB - BACKGROUND: Intracranial tumors such as meningiomas express steroid hormone receptors but little is known regarding progesterone receptor (PR) in astrocytic tumors. The authors evaluated expression of PR in 86 astrocytic tumors in relation to tumor proliferative potential. METHODS: Paraffin embedded tumor sections were stained with polyclonal antiprogesterone antibody by the peroxidase antiperoxidase method and with monoclonal MIB-1-Ki-67 antibody by avidin-biotin complex immunohistochemistry. RESULTS: Sixty-three of the 86 astrocytic tumors (73%) showed positive PR immunoreactivity. PR expression was observed in 4 of 9 pilocytic astrocytomas, 13 of 24 Grade 2 astrocytomas, 15 of 20 anaplastic astrocytomas, and 31 of 33 glioblastomas. In addition to the tumor cells, cells of microvascular endothelial proliferation and the smooth muscle of tumor vessel walls were frequently PR positive. Glioblastomas had a significantly higher percentage of PR positive cells compared with anaplastic (P < 0.0008) and low grade (P < 0.0001) astrocytomas. Patients with PR positive astrocytomas were of an older age than patients with PR negative astrocytomas (48.71 +/- 21.95 years vs. 37.09 +/- 24.69 years; P < 0.04). The mean Ki-67 labeling index (LI) was significantly higher in the high grade (3-4) astrocytomas compared with low grade (1-2) astrocytomas (P < 0.0001). PR positive astrocytic tumors had higher Ki-67 LI than PR negative tumors. PR expression was not correlated with tumor recurrence and patient survival. CONCLUSIONS: The current study suggests that PR in the astrocytic tumors correlates with histologic grade and PR may participate in the growth of these tumors and tumor angiogenesis. The measurement of PR in these tumors may indirectly represent tumor growth potential. PMID- 9392335 TI - Phase II trial of 5-fluorouracil, interferon-alpha and continuous infusion interleukin-2 for patients with metastatic renal cell carcinoma. AB - BACKGROUND: This study was designed to evaluate the efficacy and toxicity of the combination of 5-fluorouracil, interferon-alpha, and interleukin-2 for patients with metastatic renal cell carcinoma. METHODS: Previously untreated patients with a Zubrod performance status of < or =2; adequate cardiac, pulmonary, and renal function; and absence of brain metastases were eligible. One course of therapy was 28 days. 5-fluorouracil was administered at a dose of 600 mg/m2/day as a continuous infusions on Days 1-5. Interleukin-2 also was administered as a continuous infusion on Days 1-5 at a dose of 2 million Roche U/m2/day. Interferon alpha was given as a daily subcutaneous injection of 4 million U/m2/day. RESULTS: Fifty-five patients were enrolled in the trial and 52 were evaluable for response. All patients experienced fever and flu-like symptoms. Grade 3 or 4 nonhematologic toxic effects included hypertension (48%), dermatitis (12%), stomatitis (11%), and altered mental status (9%). There was one toxic death. Four complete responses and 12 partial responses were observed for a total response rate of 31% (95% confidence interval, 18-46%). The survival of responding patients was significantly better than that of nonresponding patients. The improvement in survival was even more significant when comparing patients with at least stable disease with those who progressed through treatment. CONCLUSIONS: The three-drug combination described in this study demonstrates activity. However, it appears to be more toxic than other regimens with similar response rates and cannot be recommended for standard practice. Changing the interleukin-2 route to subcutaneous administration may permit more continuous administration with less toxic effects. PMID- 9392337 TI - Pleomorphic xanthoastrocytoma: DNA flow cytometry and outcome analysis of 12 patients. AB - BACKGROUND: Pleomorphic xanthoastrocytoma (PXA) is an astrocytic tumor occurring primarily in childhood and adolescence with some malignant histologic features but a relatively slow clinical course. However, some tumors progress more rapidly and can undergo malignant degeneration. The authors attempted to determine whether various histologic features or tumor cell proliferative indices might help identify lesions at risk for early progression and distinguish PXAs from malignant gliomas. METHODS: In a retrospective study of 12 patients with PXA, the tumor's histologic features and DNA flow cytometric parameters were compared with their clinical course. DNA flow cytometry values for the S- and G2-phase of the PXAs also were compared with control group samples of malignant and low grade astrocytomas. RESULTS: Of the 12 tumors at initial diagnosis, 5 were considered typical PXAs whereas 7 had some atypical features (4 with paucity of reticulin fibers, 1 with focal necrosis, and 2 with both atypical reticulin and focal necrosis). During the follow-up period (range, 3.75-11 years; mean, 6.8 years), 2 patients had recurrences; 1 atypical reticulin PXA progressed to glioblastoma after 6.5 years and the 1 tumor with focal necrosis recurred at 6 months and again at 2 years with typical histologic features. DNA flow cytometry parameters of the typical PXA group were similar to values for malignant astrocytoma and significantly higher than values for control specimens of low grade astrocytomas. There were no distinctive DNA flow cytometric features that could distinguish this last tumor from others with a more benign clinical course. CONCLUSIONS: Measurements of the S-phase and G2-phase obtained from DNA flow cytometry and atypical histologic features cannot reliably identify PXA patients at risk for early progression and overall are significantly higher than values obtained for low grade gliomas. Therefore, frequent (i.e., two to three times per year) postoperative clinical and radiologic examinations are necessary to judge the appropriateness of adjuvant therapy in patients with PXA. The paradox of slow growth but DNA flow cytometry consistent with aggressive malignant lesions may represent a cell-cycle arrest mechanism in these lesions that could be verified in subsequent studies. PMID- 9392338 TI - Primary non-Hodgkin's lymphoma of the lacrimal sac: a case report and a review of the literature. AB - BACKGROUND: Primary non-Hodgkin's lymphoma of the lacrimal sac is extremely rare, with most reported cases representing secondary involvement of a systemic malignancy. METHODS: The clinical record of a 70-year-old female who presented with epiphora and swelling of the lacrimal sac area is described. A review of the literature of patients with primary lacrimal sac lymphoma also is presented. RESULTS: Computed tomography demonstrated a lacrimal sac mass involving the nasolacrimal canal. Histopathologic examination of a biopsy specimen revealed a diffuse large cell lymphoma. Using immunohistologic staining, the tumor was a B cell type, similar to those described in the literature. The patient was treated with 50 gray of irradiation and chemotherapy with cyclophosphamide, doxorubicin, vincristine, and prednisone. The patient remained free of lymphoma 26 months after initial treatment. An ocular examination was unremarkable except for epiphora. CONCLUSIONS: Radiotherapy and/or chemotherapy can treat localized lymphoma of the lacrimal sac successfully. PMID- 9392339 TI - Characteristics of differentiated thyroid carcinoma in children and adolescents with respect to age, gender, and histology. AB - BACKGROUND: Because of its rarity there have been only a few detailed studies on differentiated thyroid carcinoma (DTC) in children. The current investigation was undertaken to assess the characteristics of DTC with respect to age, gender, and histology in children and adolescents. METHODS: In a questionnaire-based survey, data from 114 children and adolescents with DTC (age range, 3-18 years) was collected from 65 clinical institutions in Germany. Characteristics of 80 females and 34 males were evaluated and the influence of age, gender, histology, multicentric growth, tumor stage, and lymph node involvement on distant metastases was tested using multivariate discriminant analysis. Comparison between groups was performed using the Student's t test and chi-square test. Correlation between incidence and age was assessed by linear regression analysis. RESULTS: The overall incidence of thyroid carcinoma in females was higher than in males, with a peak of female/male ratio occurring at puberty. The incidence of DTC correlated with age in females < 16 years (correlation coefficient [r] = 0.84; P = 0.0006), which was more pronounced in children with papillary thyroid carcinoma (PTC) (r = 0.83; P = 0.006) but not in those with follicular thyroid carcinoma (FTC) (r = 0.20; P = 0.16). FTC was associated with less advanced disease (P = 0.009), fewer lymph nodes involved (P = 0.007), and fewer metastases (P = 0.02) compared with PTC. Males tended to have a higher risk for distant metastases. However, statistical analysis failed to reach a level of significance (P = 0.08). Multivariate analysis revealed tumor stage as the only powerful factor (P = 0.02) correlated with distant metastasis. CONCLUSIONS: The incidence of PTC shows a marked increase in females with the highest female/male ratio occurring at puberty. Childhood thyroid carcinoma frequently is associated with lymph node involvement, distant metastases, and extrathyroidal tumor infiltration. In children FTC appears to be less aggressive than PTC. Advanced local-regional extension stage appears to be the most powerful factor influencing the risk for distant metastases in children. PMID- 9392341 TI - The National Cancer Data Base: report on kidney cancers. The American College of Surgeons Commission on Cancer and the American Cancer Society. AB - BACKGROUND: The National Cancer Data Base (NCDB) examining current time trends (1993) in stage of disease, treatment patterns, and survival of patients with kidney cancer are reported. METHODS: Five calls for data have yielded a total of 3,700,000 cancer cases for the years 1985 through 1993, including 8140 kidney cancer cases in 1988 and 10,617 in 1993 from hospital cancer registries across the U.S. These data represent 36% and 39% of all cases of kidney cancers diagnosed in the U.S. in 1988 and 1993, respectively. RESULTS: Three trends were observed. 1) Stage II disease is being diagnosed with increasing frequency. 2) There has been an overall increase in the frequency with which surgery alone is utilized as the primary treatment for patients with Stage I, II, and III disease; however, surgical treatment of patients with Stage IV kidney cancer has declined. 3) Partial nephrectomy rather than total nephrectomy is performed with increasing frequency as surgical treatment of patients with Stage I renal carcinoma. CONCLUSIONS: The NCDB data have important implications for analyzing cancer treatments and outcomes in the U.S. These data suggest that kidney cancers are being diagnosed at an earlier stage and with greater precision. As a result, changes in surgical practice are apparent in the treatment of patients with lower stage disease (Stage I and II). Stage for stage, surgery remains the most effective form of treatment for patients with kidney cancer. PMID- 9392342 TI - Infective endocarditis. PMID- 9392343 TI - Novel feature of infection in diabetic compromized hosts. PMID- 9392344 TI - Idiopathic CD4+ T-lymphocytopenia. PMID- 9392345 TI - Coronary spasm: clinical features and pathogenesis. AB - Coronary artery spasm (coronary spasm) is an abnormal contraction of an epicardial coronary artery resulting in myocardial ischemia and its incidence is relatively high in Japanese as compared with Caucasians. Coronary spasm occurs most often from midnight to early morning when the patient is at rest and it is usually not induced by exercise in the daytime. Coronary spasm can be induced by acetylcholine, an endothelium-dependent vasodilator which causes vasodilatation in the normal coronary artery. Spasm artery is hyperresponsive to the vasodilator effect of nitroglycerin, an nitric oxide (NO) donor and is deficient in NO activity. The major risk factor for coronary spasm is cigarette smoking. Coronary spasm can be a cause of not only variant angina but also ischemic heart disease in general, including unstable angina, acute myocardial infarction and sudden ischemic death. PMID- 9392346 TI - Involvement of peribiliary glands in primary sclerosing cholangitis: a histopathologic study. AB - We examined the histological changes of the peribiliary glands (PBGs), a hitherto pooly recognized anatomical element around the biliary tree, in 7 cases of primary sclerosing cholangitis (PSC). These glands showed proliferation, and nonspecific inflammation with lymphoplasmacytic infiltration, fibrosis, and destruction. In addition, there were cystic lesions around the bile ducts, and they were considered to reflect dilatation of the PBGs. These changes were found around the intrahepatic and extrahepatic bile ducts in the cases examined. It is of interest that changes in the PBGs tended to correlate with the inflammatory changes of the bile duct wall itself, though 2 cases showed changes in the duct walls and PBGs unrelated to their distribution along the biliary tree. These findings suggest that the PBGs are also a target structure in addition to the bile ducts themselves in PSC. PMID- 9392347 TI - Effect of respiratory rate on respiratory patterns in patients with chronic obstructive pulmonary disease. AB - We enrolled 22 patients with chronic obstructive pulmonary disease (COPD) and 20 normal subjects as controls. Using a hot-wire flow meter, we obtained tidal volume (VT), duty ratio (Ti/Ttot), and mean inspiratory flow (VT/Ti) as measures of respiratory pattern at two different respiratory rates; 0.5 Hz and 1.5 Hz. At 0.5 Hz, there were significant differences in Ti/Ttot and VT/Ti. At 1.5 Hz, patients with COPD had significantly lower values of VT and VT/Ti. Furthermore, we calculated the change ratios from 0.5 Hz to 1.5 Hz in these parameters as new parameters of respiratory pattern change by respiratory rate. VT0.5/1.5 and VT/Ti0.5/1.5 significantly correlated with FEV1.0/FVC. The findings suggest that the parameters of the respiratory pattern may change depending on respiratory rates, and that the ratios of those parameters obtained at two different rates could be helpful in diagnosing airflow obstruction. PMID- 9392348 TI - Ventilatory responses and subjective sensations during arm exercise and hypercapnia in patients with lower-cervical and upper-thoracic spinal cord injuries. AB - We measured the ventilatory responses and subjective sensations during arm exercise in patients with lower cervical and upper thoracic spinal cord injuries in order to evaluate the effects of chest wall deafferentation on these responses. Visual analog scales with verbal descriptors were used to quantify respiratory sensations of different affectional qualities. Patients as well as normal subjects reported stronger respiratory sensations upon CO2 rebreathing as compared to during arm exercise with an equivalent minute ventilation (p<0.05). There were no qualitative nor quantitative differences in the respiratory sensations during CO2 rebreathing between the patients and normal subjects. However, patients with spinal cord injuries showed a higher minute ventilation and a lower end-tidal PCO2 during incremental arm exercises (p<0.01), and thus tended to hyperventilate. We conclude that chest wall afferent denervation does not contribute significantly to the perception of breathlessness in patients with spinal cord injuries. PMID- 9392350 TI - Multiple gastric carcinoids and pituitary adenoma in type A gastritis. AB - A 48-year-old male with type A atrophic gastritis developed multiple gastric carcinoids and a pituitary adenoma. Laboratory tests revealed high levels of serum gastrin and growth hormone (GH). He underwent subtotal gastrectomy, resulting in a return of the previously elevated gastrin level to normal. Serum GH concentration remained high. Three months after the surgery, the pituitary tumor, composed greatly of GH-immunoreactive cells, was partially removed. Since hypergastrinemia plays a pivotal role in gastric carcinoid formation and induces GH-releasing factor (GHRH) release resulting in GH-producing pituitary tumor formation, GH-producing pituitary adenoma might be a clinical manifestation in type A gastritis. PMID- 9392349 TI - Detection of polyanion-restricted anti-histone antibodies in patients with systemic lupus erythematosus. AB - The nature of the antibodies responsible for lupus erythematosus (LE) cells in systemic lupus erythematosus (SLE) remains obscure. We examined whether polyanion restricted anti-histone antibodies were present in serum of patients with SLE using Western blotting analysis. Dextran sulfate or alginate was used as a polyanion compound in place of DNA. Antibodies which recognized dextran sulfate histone complexes were present in serum of patients with SLE (17/34, 50%). These antibodies were detected in most SLE patients positive for LE cells (17/18, 94%) but not in those negative for LE cells or in patients with other collagen diseases. Similar results were obtained using alginate-histone complexes as antigens for Western blotting analysis. The antibodies to dextran sulfate-histone or alginate-histone complexes in serum of SLE patients were completely absorbed by treatment of serum with DNA-histone complexes, while they were unaffected by treatment with DNA only. The presence of antibodies to free histones and dextran sulfate-histone complexes did not seem to be related to the titer of anti-single stranded DNA antibody and anti-double stranded DNA antibody. We demonstrated the presence of polyanion-restricted antibodies in SLE, which may be responsible for the LE factor. PMID- 9392351 TI - Portal-hepatic venous shunt through a portal aneurysm complicated by hepatic encephalopathy and pulmonary hypertension. AB - We report a rare case of portal-hepatic venous shunt through an enormous portal aneurysm complicated by pulmonary hypertension. A 66-year-old woman was admitted to our hospital for hepatic encephalopathy. Chest roentgenography revealed pulmonary hypertension. Computed tomography and ultrasound examination demonstrated a shunt between the portal and hepatic veins through an enormous portal aneurysm. The diagnoses of portal-hepatic venous shunt and pulmonary hypertension were confirmed by hepatic venous catheterization and cardiac catheterization. Pulmonary hypertension might result from the effects of vasoconstrictive agents, which should be metabolized by the liver in normal subjects, passing through the intrahepatic shunt into the lung. PMID- 9392352 TI - Long-term continuous intravenous infusion of prostacyclin for severe primary pulmonary hypertension. AB - A patient suffering from severe symptomatic primary pulmonary hypertension (PPH) underwent long-term intravenous prostacyclin therapy; the first time for such treatment in Japan. A 26-year-old male had experienced gradually progressive dyspnea for about one year. Despite conventional therapy he suffered repeated syncopal attacks. However, after receiving a permanent central venous access device and a portable infusion pump, he recovered fully and was discharged. This remedy seems to be promising for PPH as has already been proven in Europe and North Americas, although in Japan it is not as yet commercially available and some problems still need to be resolved. PMID- 9392353 TI - Tricuspid valve endocarditis in a non-drug addict associated with peliosis hepatis. AB - A 31-year-old woman was admitted because of persistent remittent fever. Tricuspid valve endocarditis due to Staphylococcus aureus was identified as the cause of fever. The patient had no history of intravenous drug abuse, oral contraceptives or predisposing cardiac disease. Huge hepatomegaly was found without any signs of congestive heart failure. Liver enzyme abnormalities were not detected throughout the entire course of therapy. The liver biopsy specimen revealed peliosis hepatis. Treatment with panipenem/betamipron was successful, although recurrent septic pulmonary embolism occurred. The cause of the huge hepatomegaly encountered in the present case may be attributable to peliosis due to severe infection. PMID- 9392354 TI - Cushing's syndrome due to bilateral adrenocortical adenomas with different pathological features. AB - A 48-year-old woman with Cushing's syndrome due to bilateral adrenocortical adenomas is reported. The patient presented with a typical Cushingoid appearance. The serum cortisol level was elevated with loss of the diurnal rhythm and the plasma adrenocorticotropic hormone (ACTH) level was undetectable. Dynamic testing showed no suppression of urinary 17-OHCS by high-dose dexamethasone and no stimulation by metyrapone. An abdominal computed tomography (CT) scan showed bilateral adrenal tumors. Bilateral adrenalectomy was performed. The right adrenal gland contained a tumor that was encapsulated and consisted mainly of compact cells. The surrounding cortex was atrophic. The left adrenal gland contained an encapsulated tumor composed predominantly of clear cells. There were numerous small adrenocortical nodules in the surrounding cortex. Immunohistochemical analysis of steroidogenic enzymes (P450scc, 3beta-HSD, P450c21, P450c17 and P450c11) was performed. Immunoreactivity of all the enzymes was intense in the compact cells of the right adrenocortical adenoma, while the adjacent non-neoplastic cortex was negative for the enzymes. In the left adrenal tumor, the immunoreactivity of 3beta-HSD was intense, while that of P450c17 was weak. In the adrenocortical nodules, 3beta-HSD activity was sporadically observed. G protein genes encoding Gs alpha and Gi2 were examined for activating mutations at codons 201 and 227 (Gs alpha) and codons 179 and 205 (Gi2 alpha) in the bilateral adrenal tumors, but no mutations were found. The bilateral adenomas of this patient showed marked differences in microscopic and immunohistochemical studies, suggesting that the capacity of steroidogenesis differs between the right and left tumors. PMID- 9392355 TI - Pyogenic clavicular osteomyelitis associated with disseminated intravascular coagulation and acute renal failure in a patient with non-insulin-dependent diabetes mellitus. AB - Pyogenic osteomyelitis is often accompanied by diabetes, but the disease in the clavicula has rarely been reported. We describe an unusual case of a 53-year-old man with poorly controlled non-insulin-dependent diabetes mellitus who presented with pyogenic clavicular osteomyelitis and developed DIC and acute renal failure. A 67Ga scintigram revealed an abnormal accumulation of the isotope in the right clavicula, where magnetic resonance imaging (MRI) showed inflammatory changes. This suggests that a 67Ga scintigram and MRI are of clinical value for the early diagnosis of the disease. Antibiotic chemotherapy with gamma-globulin and gebexate mesilate, and hemodialysis almost cured his serious condition. PMID- 9392356 TI - Massive adrenal hemorrhage secondary to metastasis of lung cancer. AB - Hemorrhagic adrenal metastasis from lung cancer is extremely rare, although adrenal involvement is common in widely disseminated cancer. We report a case of massive adrenal hemorrhage secondary to metastasis of lung cancer. A 47-year-old female was treated by left upper lobectomy and mediastinal lymph node resection for an adenocarcinoma with intrapulmonary metastasis in the left upper lobe. Eight months later, she presented with right flank and back pain, and abdominal ultrasonography and computed tomography showed a right solitary adrenal tumor with massive hemorrhage. The tumor was not resectable and partially responded to chemotherapy. A massive adrenal hemorrhage, secondary to metastasis of lung cancer, presents with nonspecific clinical signs and symptoms. In lung cancer patients with an acute flank or back pain, hemorrhagic adrenal metastasis should be considered in the differential diagnosis. PMID- 9392357 TI - Acute lymphoblastic leukemia with isolated adrenocorticotropic hormone deficiency. AB - A 57-year-old female was admitted to our hospital because of Philadelphia chromosome (Ph)-positive acute lymphoblastic leukemia (ALL). On admission, disturbance of consciousness and hyponatremia were recognized. The patient's endocrinological data showed low levels of adrenocorticotropic hormone (ACTH) (less than 5 pg/ml) and cortisol (5.9 microg/dl). Other anterior pituitary hormones were normal. Plasma ACTH and cortisol did not respond to the corticotropin releasing factor (CRF) test. A diagnosis of isolated ACTH deficiency was made. This is a rare case of isolated ACTH deficiency complicated with hematological malignancies. PMID- 9392359 TI - Facial nerve enhancement on gadolinium-DTPA in a case with neurosarcoidosis. AB - In a case of neurosarcoidosis with bilateral facial nerve palsy and hydrocephalus, contrast-enhanced magnetic resonance imaging (MRI) study and angiotensin converting enzyme (ACE) activities in cerebrospinal fluid (CSF) were valuable for the diagnosis and the follow up. Facial nerve lesions were demonstrated on gadolinium-DTPA enhanced MRI. The disappearance of enhancement was concomitant with the amelioration of facial nerve palsy after corticosteroid therapy. PMID- 9392358 TI - Idiopathic CD4+ T-lymphocytopenia with Bowen's disease. AB - A 39-year-old man with Bowen's disease was troubled with multiple molluscum contagiosum over the trunk and lower extremities. Subsequently oral candidiasis was complicated. Laboratory examination revealed lymphocytopenia and a decrease in the CD4/CD8 ratio. His CD4+ T-lymphocyte count was only 187 cells/microl one time and 222 cells/microl another time. No evidence for human immunodeficiency virus (HIV) infection was found. He had no family history of immunodeficiencies. PMID- 9392361 TI - Spontaneous dissection associated with proximal vertebral artery anomaly. AB - A 47-year-old man was admitted because of acute lateral medullary syndrome with severe posterior cervical pain. Cerebral angiography was performed three hours after the onset, which demonstrated that two arteries branched separately from the right subclavian artery, ran upward and formed a single right vertebral artery (VA). One of the two arteries showed both stenosis and luminal dilatation. We thought the structure of these arteries was proximal vertebral artery anomaly and diagnosed him as having dissection of the vertebral artery. We consider that the proximal vertebral anomaly may be a risk for spontaneous VA dissection. PMID- 9392360 TI - Sensory conduction study of cisplatin neuropathy: preservation of small myelinated fibers. AB - We report a patient with peripheral neuropathy caused by cisplatin for the treatment of testicular tumor. Routine studies of nerve conduction and somatosensory evoked potentials demonstrated large myelinated fiber neuropathy suggesting ganglioneuronopathy. We also performed a CO2 laser evoked potential study, and found that small myelinated fibers, which are related to pain sensation, were well preserved in this patient. PMID- 9392362 TI - Pulmonary fibrosis associated with polymyalgia rheumatica. AB - Polymyalgia rheumatica (PMR) and exacerbated pulmonary fibrosis presented concurrently in a 69-year-old woman with a 5-year history of idiopathic interstitial pneumonia. The radiographic and histological examinations suggested usual interstitial pneumonia (UIP) as a more likely diagnosis. Corticosteroid therapy resulted in relief of the patient's muscle symptoms and improvement in the functional and radiographical signs of the pulmonary fibrosis. The final diagnosis was pulmonary fibrosis associated with PMR, because PMR is believed to be one of the causes contributing to interstitial lung diseases. PMID- 9392363 TI - Myeloperoxidase-antineutrophil cytoplasmic antibody-associated glomerulonephritis with membranous nephropathy in remission. AB - A 47-year-old woman developed pulmonary hemorrhage and an increase in proteinuria during remission of membranous nephropathy. Renal biopsy revealed crescentic glomerulonephritis. She also had a high perinuclear antineutrophil cytoplasmic antibody level, so a diagnosis of myeloperoxidase-antineutrophil cytoplasmic antibody-associated glomerulonephritis was made. After immunosuppressive therapy was started, the pulmonary hemorrhage resolved and her proteinuria decreased. Renal biopsy was repeated after treatment and showed histological improvement. This case suggests that there may be a relationship between membranous nephropathy and myeloperoxidase-antineutrophil cytoplasmic antibody-associated glomerulonephritis. PMID- 9392364 TI - Reuse of a Japanese familial amyloidotic polyneuropathy patient's liver for a cancer patient: the domino liver transplantation procedure. PMID- 9392365 TI - New packing materials for protein chromatography. AB - This review describes new packing materials designed for protein chromatography, covering advances in base supports and stationary phases. Base supports are classified according to their chemical composition. Since most separation media are bead shaped, typical procedures used for their preparation are also presented. In order to provide matrices combining improved chemical stability and chromatographic performances, composite materials continue to be developed, including bonded stationary phases, pore composites and mixed carriers. The different approaches to their preparation are described and characteristics that play a major role in the chromatographic process are discussed. Recently introduced materials and some of their applications under non-denaturing conditions in the different chromatographic modes are also presented. PMID- 9392366 TI - Protein separation using non-porous sorbents. AB - This article overviews the development of non-porous sorbents having small particle diameters which have proven effective for rapid analysis and micropreparative separation of proteins by liquid chromatography. Much attention is given to the preparation and application of silica- and polystyrene-based non porous packings for various chromatographic modes, especially affinity chromatography. Modeling works on the prediction and parameter estimation for the dynamics of protein adsorption using non-porous sorbents are reviewed and briefly described. To conclude this review, future prospects of the application of non porous sorbents are also presented. PMID- 9392367 TI - Permeable packings and perfusion chromatography in protein separation. AB - The use of permeable packings in perfusion chromatography for protein separation is reviewed. Mass transport mechanisms in large-pore materials include forced convection in addition to diffusive transport. The key concept in perfusion chromatography is the "augmented" diffusivity by convection which explains the improved efficiency of perfusive packings compared with conventional supports. An extended Van Deemter equation has to be applied when calculating the height equivalent to a theoretical plate (HETP) of chromatographic columns with flow through particles. It is shown that the effect of forced convective flow in pores is to drive the separation performance between diffusion-controlled and equilibrium limits. A methodology to understand mass transfer mechanisms in permeable packings is proposed. Experimental results for protein separation by high-performance liquid chromatography in new packing media are discussed. Simulated moving bed technology is addressed. PMID- 9392368 TI - Electrophoresis gel media: the state of the art. AB - Some unique events have occurred in the last few years which might revolutionize the field of polyacrylamide gel electrophoresis. While it was widely recognized that such matrices could normally be cast with a small pore size distribution, typically of the order of a few nanometers diameter (for protein sieving), recent developments suggest that "macroporous" gels could also be produced in the domain of polyacrylamides. If constraints to chain motion are imposed during gel polymerization, large-pore structures can be grown. Such constraints can originate either from low temperatures or from the presence of preformed polymers in the gelling solution; in both cases, the growing chains are forced to "laterally aggregate" via inter-chain hydrogen bond formation. Upon consumption of pendant double bonds, such bundles are frozen in the three-dimensional space by permanent cross-links. As an additional development, a novel photopolymerization system is described, comprising a cationic dye (methylene blue) and a redox couple (sodium toluene sulfinate, a reducer, and diphenyliodonium chloride, a mild oxidizer). Methylene blue catalysis is characterized by a unique efficiency, ensuring >96% conversion of monomers, even in hydro-organic solvents and in the presence of surfactants, which normally quench or completely inhibit the persulphate-driven reaction. In addition, methylene blue-sustained photopolymerization can be operated in the entire pH 3 10 interval, where most other systems fail. Perhaps the most striking novelty in the field is the appearance of a novel monomer (N-acryloylaminopropanol, AAP) coupling a high hydrophilicity with a unique resistance to alkaline hydrolysis. Given the fact that a poly(AAP) matrix is 500 times more stable than a poly(acrylamide) gel, while being twice as hydrophilic, it is anticipated that this novel chemistry will have no difficulties in replacing the old electrophoretic anticonvective media. The review ends with a glimpse at novel sieving media in capillary zone electrophoresis: polymer networks. Such media, by providing an almost infinite range of pore sizes, due to the absence of a rigid support, allow sieving mechanisms to be operative over a wide interval of particle sizes, even up to genomic DNA. Viscous solutions of polymer networks, made with the novel poly(AAP) chemistry, allow repeated use of the same separation column, well above 50 injections. Silica-bound poly(AAP) chains provide effective quenching of electroosmosis and >200 analyses by isoelectric focusing. PMID- 9392369 TI - Isoelectric focusing in immobilized pH gradients: an update. AB - The latest trends on isoelectric focusing (IEF) in immobilized pH gradients (IPG) are here reviewed. The major advances on IPG technologies have been made when interfacing this technique with sodium dodecyl sulfate-polyacrylamide gel electrophoresis to produce two-dimensional (2-D) maps. Previous 2-D maps were routinely performed using conventional IEF as a first dimension, which typically resulted in poor reproducibility of spot position. With IPGs, correlation between experimental and calculated protein pI values is as good as +0.01 to 0.02 pH units. A new software has also been released, permitting easy calculation and optimization of linear, concave and convex exponential gradients, even in very complex recipes utilizing all ten Immobiline chemicals. It has also been proven that IPGs can be interfaced with mass spectrometry, thus obtaining a novel 2-D map with the best of pI measurements in the first dimension coupled with the best of mass determination in the second dimension. Recently, it has been shown that IPGs can be exploited to charter forbidden grounds, with the creation of non linear pH gradients covering the extreme alkaline pH 10-12 gradient. In such basic regions, excellent steady-state patterns of histones and subtilisin mutants have been reported. Different families of histones could be mapped not only in this pH 10-12 interval, but also in 2-D maps exploiting this very alkaline gradient in the first dimension. Although the IPG technique is now a trouble free, user-friendly technique, some annoying artefacts, producing severe protein smears and precipitation, were very recently reported, but found to be linked to some commercial Immobiline preparations containing up to 5% oligomers. Better quality control on the part of the company producing such chemicals should eliminate even this last source of troubles. PMID- 9392370 TI - Current trends in capillary isoelectric focusing of proteins. AB - Isoelectric focusing (IEF) in thin capillaries is reviewed here. After an introduction on the genesis and chemistry of the carrier ampholyte buffers, different approaches to IEF are discussed and evaluated. The classical approach consists on IEF under conditions of suppressed electroosmotic (EOF) flow, usually obtained by covalently bonding hydrophilic polymers to the inner capillary wall. The other approach consists of IEF in dynamically (and partially) coated capillaries, so as to allow a reduced EOF flow to coexist with the IEF process, so that focusing and transport of the train of stacked bands occurs simultaneously. The various experimental parameters: focusing, elution and detection steps, pI measurements, as well as typical drawbacks, such as isoelectric precipitation are evaluated. The review ends with some examples of analytical separations, at the moment mostly limited to focusing of native hemoglobins (normal and point mutants). These separations are compared with those obtained by slab-gel IEF and in immobilized pH gradients. PMID- 9392371 TI - Protein purification in multicompartment electrolyzers with isoelectric membranes. AB - Preparative purification of proteins under isoelectric conditions is reviewed, with particular regard to novel equipment, a multicompartment electrolyzer with isoelectric membranes, which can capture any desired protein into an isoelectric trap as the sole, ultra-pure component. This novel machine is based on the Immobiline chemistry, i.e. the novel generation of non-amphoteric buffers, based on the chemistry of acrylamides, which can be insolubilized onto polyacrylamide supports. After a description of the instrument and of its performance, a number of protein purification protocols are described, leading to truly homogeneous (by the most stringent criterion of surface charge) protein fractions. Such a high charge purity has been found to be often a fundamental prerequisite for the growth of protein crystals. Interfacing the electrolyzer with mass spectrometry has permitted the decoding of the structure of minor components generated from a parental molecule, especially ones having a higher pI. It was found that these species were often generated either by proteolytic cleavage or by the formation of a trisulphide bridge between two Cys residues. A unique application of the electrolyzer is finally described: its use as an immobilized enzyme reactor under an electric field. The performance of this reaction is outstanding, in that the kinetic parameters of the immobilized enzyme are identical to those of a free enzyme form. PMID- 9392372 TI - Factors affecting protein interaction at sorbent interfaces. AB - Interactions between surfaces and macromolecules are the fundamentals in separation and detection of diverse solutes. In this very brief review the central aspects of protein-surface interactions are discussed with the intention of identifying the important factors influencing such processes and placing them in relation to the established knowledge in this field. Some perspectives of new techniques related to scanning probe microscopy for studying interactions at the nanometer level are also discussed. PMID- 9392373 TI - Rules relating electrophoretic mobility, charge and molecular size of peptides and proteins. AB - The absence of supporting media in free solution high-performance capillary electrophoresis (HPCE) makes it an ideal system for the study of the relationship between electrophoretic mobility (mu(em)) and the molecular size and charge of proteins and peptides. In this review, the theory of electrophoresis, developed for rigid, insulating, spherical particles, is modified to develop models for the electrophoretic behaviour of proteins and peptides. For a given set of experimental conditions, mu(em) of a protein/peptide is proportional to its charge (q) and is inversely proportional to its Stoke's radius (r). Furthermore, mu(em) is most sensitive to changes in q and, as a consequence, the reliability of equations relating mu(em) to protein/peptide q and r is dependent upon the accurate calculation or determination of q. For convenience, q and r of proteins and peptides are generally expressed in terms of calculated valence (Zc) and molecular mass (M), respectively, both of which can be determined from the amino acid sequence of the protein/peptide. However, the calculation of q using Zc is made more complex by the effects of electrostatic charge suppression, such that Zc is an overestimation of actual charge. Charge suppression becomes increasingly significant as the protein/peptide charge increases, such that, for peptides, the relationship between q and Zc can be approximated by a logarithmic function. The mu(em) for peptides, therefore, can be approximated by the equation: mu(em) = ln(Zc + 1)/K Ms where s varies between 1/3 and 2/3, and K is a constant that is valid for a particular set of experimental conditions. The rather simplistic compensation for charge suppression in this equation is inadequate for proteins where the magnitude of charge suppression is greater and the mechanisms are more complex. For proteins, the relationship suggested for the prediction of mu(em) from Zc and M is: mu(em) = Zc/KFzMs where s again varies between 1/3 and 2/3 and Fz is a pH-independent proportionality factor defined as the quotient, Zc/Za, with Za being actual protein valence. The factor Fz can be determined empirically, however, it is valid only for the particular set of experimental conditions under which it is determined. For peptides, the mass exponent, s, approaches 1/3 when the peptides have high charge densities and open structures. However, s approaches 1/2 for peptides with lower charge densities that are capable of more randomized motion during electrophoresis. Finally, s approaches 2/3 for proteins, suggesting that the frictional forces acting on a protein undergoing electrophoretic motion are proportional to the surface area of these larger, more rigid, structures. In conclusion, the development of relationships between mu(em), M and Zc for peptides and proteins offers a powerful tool, not only for predicting electrophoretic mobility, but also for optimising HPCE separations, studying structural modifications (e.g. phosphorylation, glycosylation, deamidation, etc.), and for the investigation of surface charge characteristics and conformation. PMID- 9392374 TI - Peptide analysis by capillary (zone) electrophoresis. AB - In this review various aspects concerning the application of capillary (zone) electrophoresis for peptide analysis will be critically examined. First, the basic instrumental requirements of CE apparatus and the strategies employed to enhance sensitivity in the analysis of underivatized sample are described. Multidimensional separative techniques of complex peptide mixtures that use CE as final step and the coupling of CE with mass spectrometry are subsequently discussed. A theoretical section describes the relationships existing between peptide mobility and the pH of the separation buffer. These relationships evidence that proton dissociation constants and Stokes radius at different protonation stages can be calculated by measuring the electrophoretic mobility at different pH values. Investigation of peptide mobility dependence on pH allows us to establish the optimum conditions, in terms of resolution, for peptide separation. Subsequently, a critical discussion about semiempirical models predicting peptide mobility as a function of chemico-physical peptide properties is presented. A section is devoted to the description of principles of peptide affinity capillary electrophoresis, underlining the similarity with peptide proton interaction. CE separations performed in aquo-organic solvents are also critically discussed, showing the good performance obtained by using water-2,2,2 trifluoroethanol solutions. Finally, selected CE applications for the determination of peptide chemico-physical properties and conventional analysis, like peptide mapping, are reported. PMID- 9392375 TI - Labeling reactions applicable to chromatography and electrophoresis of minute amounts of proteins. AB - Chromatography and electrophoresis have become extremely valuable and important methods for the separation, purification, detection and analysis of biopolymers and HPLC/HPCE may become the premier, preferable approaches for both qualitative and quantitative analyses of most proteins, especially from recombinant materials. This includes smaller peptides, polypeptides, proteins, antibodies and all types of protein or antibody-conjugates (antibody-enzyme, protein-fluorescent probe, antibody-drug and so forth). This entire Topical Issue of Journal of Chromatography emphasizes the application of chromatography and electrophoresis to protein analysis. This particular review deals with approaches to the selective tagging or labeling of proteins at trace (minute) levels, again using either chromatography or electrophoresis, with the emphasis on modern HPLC/HPCE methods and approaches. We discuss here both pre- and post-column labeling methods and reagents, techniques for realizing selective labeling of proteins or antibodies, applicable approaches to protein preconcentration in both HPLC and HPCE areas and in general, methods for improving (lowering) detection limits for proteins utilizing chemical or physical derivatization and/or preconcentration techniques. There are really two major goals or emphases in that which follows: (1) methods for selective labeling of proteins prior to or after HPLC/HPCE and (2) labeling of proteins at trace levels for improved separation-detection and lowered detection limits. We discuss here a large number of specific references related to both pre- and post-column/capillary derivatizations for proteins, as well as methods for improved detectability in both HPLC and HPCE by, for example, analyte preconcentration on a solid-phase extractor or membrane support, capillary isotachophoresis and other methods. Selective reactions or derivatizations on proteins refers to the ability to tag the protein at specific (e.g. reactive amino sites) in a controlled manner, with the products having the same number of tags all at the very same site or sites. The products are all the same species, having the same number of tags at the same locations on the protein. Selective reactions can also refer to the idea of tagging all of the protein sample at only a single, same site or at all available sites, homogeneously. PMID- 9392376 TI - Chromatography on cells and biomolecular assemblies. AB - Red cells, biomembrane vesicles, proteoliposomes and liposomes non-covalently immobilized in gel particles or beads have been used as stationary phases for biomembrane affinity analyses and ion-exchange chromatographic separation. Lipid monolayers coupled to silica beads have been utilized for membrane protein purification in detergent solution and plant cell walls for group separation of macromolecules according to size and charge. Further methodological studies are essential to implement general practical application. PMID- 9392377 TI - Buffer additives other than the surfactant sodium dodecyl sulfate for protein separations by capillary electrophoresis. AB - The different compounds utilized as additives to the electrolyte solutions employed in protein capillary zone electrophoresis (CZE) for minimizing protein capillary wall interactions, for improving selectivity and resolution and for controlling the electroosmotic flow are reviewed. The dependence of the electroosmotic flow on the different variables that can be affected by the incorporation of an additive into the electrolytic solution is discussed. A list of the most effective additives employed for protein separations by CZE is reported in Appendix A. PMID- 9392378 TI - Capillary electrophoresis procedures for serum protein analysis: comparison with established techniques. AB - Methods using automated capillary electrophoresis (CE) instrumentation are available for serum protein electrophoresis with monoclonal band quantitation, isoelectric focusing and sodium dodecyl sulphate-polyacrylamide gel electrophoresis separations. The advantages of CE over previous gel methods relate to the time and labour saved by the automated instrumentation. High pI monoclonal bands and cryoglobulin specimens can be successfully analysed by CE. However, if the CE application uses a standard company supplied kit, then the cost savings are often negated by the high cost of the kit. Improvements such as the inclusion of both a UV-Vis as well as a fluorescence detector as standard within the one commercial instrument, the production of coated IEF capillaries with a useful life of at least 100 samples, and the introduction of a capillary array into all commercial instrumentation would ensure greater use of CE within both the clinical and other protein laboratories. PMID- 9392379 TI - Chromatographic and electrophoretic methods for modified hemoglobins. AB - The discovery of the clinically important glycohemoglobin adducts and their relation to diabetes mellitus have greatly stimulated the study of other minor post-translational modifications of hemoglobin. Chromatographic and electrophoretic procedures have played an important role in these studies. Today several hemoglobin adducts are known and the formation of adducts with glucose, phosphorylated carbohydrates, urea/cyanate, aspirin, vitamins, acetaldehyde, penicillin and acetyl CoA have been described. Furthermore, new adducts, such as those observed using hemoglobin as a biochemical marker monitoring environmental, occupational and lifestyle exposures to reactive toxic chemicals are constantly being reported. This review deals with chromatographic and electrophoretic separation methods available for the study of non-enzymatic post-translational modifications of hemoglobin. Suitability, perspectives and biomedical applications are discussed. PMID- 9392381 TI - Post-translational non-enzymatic modification of proteins. II. Separation of selected protein species after glycation and other carbonyl-mediated modifications. AB - There are two strategies applicable to revealing non-enzymatic post-translational modifications of proteins; while assaying of the hydrolytically stable adducts was the subject of our previous communication [1], here we attempted to review separation technologies for the unfragmented modified proteins. There are a few standard procedures used for this purpose, namely Laemmli gel electrophoresis, different modes of gel permeation chromatography and boronate affinity chromatography. The latter approach makes use of the vicinal hydroxy groups present in glycated proteins. Some (but not all) arising adducts exhibit typical fluorescence which can be exploited for detection. In most cases fluorescence is measured at 370/440 nm for the so-called advanced glycation products or at 335/385 nm for the only so far well characterized glycation marker (pentosidine). Some indication exists that, e.g., synchronous fluorescence detection will probably in the future add to the selectivity and allow the distinction of the different adducts arising during non-enzymatic post-translational modifications (glycation). The proteins reviewed are serum albumin, collagen and lens proteins while glycation of hemoglobin is the subject of another review within the present volume. PMID- 9392380 TI - Post-translational non-enzymatic modification of proteins. I. Chromatography of marker adducts with special emphasis to glycation reactions. AB - Analytical methods for marker compounds formed during post-translational modifications of proteins are reviewed. Only adducts arising either in vivo or under in vitro conditions simulating the in vivo situations are discussed. All of these compounds stem from either the reaction of free amino groups (i.e., lysine, arginine or N-terminal amino acid). In most cases the reactive counterpart is an aldehydic moiety containing endogenous compound; however, other functional groups containing metabolites are considered as well. The main demand put upon such marker compounds is that they are stable in acid or enzymatic hydrolysis or, alternatively, can be stabilized by simple sample pretreatment (e.g., by reduction). Practically all categories of separation procedures can be applied provided that the chemical characteristics of a particular marker are adequately respected; frequently combination of two different separation procedures based on different principles must be used. Because of the low level of such marker compounds under in vivo conditions, an appropriate sample enrichment step must be involved. Emphasis is put upon the analysis of Amadori products, pentosidine (and pentosidine related compounds), pyrraline, furosine, N-carboxymethylamino acids, amino acid hydantoins and stabilized dicarbonyl intermediates. PMID- 9392382 TI - Unstable proteins: how to subject them to chromatographic separations for purification procedures. AB - The chromatographic separation of an unstable protein is often a challenge to the scientist working in the field of life sciences. Especially for the purification of sensitive enzymes, making use of conventional chromatographic techniques is difficult and frequently results in a complete loss of biological activity of the target protein. This report summarizes some general strategies that may help to keep unstable proteins in their native conformation during the rather harsh conditions of a purification procedure. In this context, a recently developed hollow fiber membrane module, suitable for performing on-line dialysis, is introduced and examples of its application to liquid column chromatography are given. Many innovative separation techniques, characterized by dramatic improvements in both performance and separation time, have recently been developed. Since the chromatographic separation of unstable proteins requires the use of modern state-of-the-art equipment and technology, emphasis is given to newly developed separation techniques such as expanded bed adsorption, perfusion chromatography, protein free flow electrophoresis and the use of tentacle gels. In addition, examples of recently published purifications of unstable proteins are discussed with respect to strategies ensuring the preservation of the native protein structure during chromatographic separation. PMID- 9392383 TI - Separation methods for glycoprotein analysis and preparation. AB - Several chromatographic methods have been developed for the isolation and characterization of glycoproteins. In these methods, affinity chromatography, a single-step method, or combined use with general chromatographic methods have now become essential for the purification of many biologically important glycoproteins, including alpha1-acid glycoprotein, immunoglobulins, ceruloplasmin and erythropoietin. On the other hand, almost all glycoproteins exhibit polymorphism associated with their glycan moieties. This feature is wide-spread and has been observed in natural as well as in recombinant DNA glycoproteins. Recently, several sophisticated techniques--such as electromigration method (high performance capillary electrophoresis) and chromatographic methods (two dimensional polyacrylamide gel electrophoresis, high-pH anion-exchange chromatography with pulsed-amperometric detection)--have been introduced for qualitative or quantitative estimation of the microheterogeneity of glycoproteins. For gaining further insight into the structure-function relations for microheterogeneity, preparative chromatographic techniques that can yield sufficient quantities of glycoprotein variants must be developed. PMID- 9392385 TI - High-performance separations in isolation and characterization of allergens. AB - The present state of the use of separation techniques in the identification and characterization of allergens and in the monitoring of the quality of allergenic preparations is critically surveyed. After a brief summary of the range of problems encountered in obtaining and in the application of allergenic preparations and of the principal physico-chemical properties of allergens, chromatographic and electromigration methods of separation of components of these systems and their combinations with immunochemical procedures are discussed, with selected examples of application to real materials. Emphasis is placed on evaluation of the most important analytical parameters, such as reliability of the results, separation efficiency and resolution, and on the most recent results in the field. PMID- 9392384 TI - Separation used for purification of recombinant proteins. AB - The purification of molecules from recombinant cells may be strongly influenced by the molecular biology of gene isolation and expression. At the beginning of the process there may be a demand for information on the minute amounts of proteins and thus for ever increasingly sensitive techniques. Purification of recombinant proteins can differ from conventional purifications in several ways, depending on the solubility of the protein, occurrence in inclusion bodies, creation of fusion proteins with tags that enable simpler purification. Sometimes a (re)naturation step is required to get a bioactive protein. On the other hand, the techniques used in separation are essentially the same as for purification from the natural source and environment. PMID- 9392386 TI - Lipoproteins: comparison of different separation strategies. AB - This review describes two chromatographic techniques for the separation of three main classes of lipoproteins (HDLs, LDLs and VLDLs) from human serum: hydroxyapatite chromatography and counter-current chromatography. The HDLs, LDLs and VLDLs were purified by the combined use of the two chromatographic techniques without prior ultracentrifugation. PMID- 9392387 TI - Analysis of protein aggregates by combination of cross-linking reactions and chromatographic separations. AB - Chemical cross-linking provides a method that covalently bridges near-neighbour associations within proteins and protein aggregates. Combined with chromatographic separations and protein-chemical methods, it may be used to localize and to investigate three-dimensional relations as present under natural conditions. This paper reviews the chemistry and application of cross-linking reagents and the development of combination experimental approaches in view of chromatographic separations and cross-linking reactions. Investigations of homooligomeric and heterooligomeric protein associations as well as conformational analysis are presented. PMID- 9392388 TI - Capillary electrophoretic immunoassays. AB - Capillary electrophoretic immunoassay (CEIA) has recently emerged as a new analytical technique. CEIA, when combined with sensitive detection methods such as laser induced fluorescence (LIF), offers several advantages over conventional immunoassays. CEIA can perform rapid separations with high mass sensitivity, simultaneously determine multiple analytes and is compatible with automation. The objective of this review is to describe the applications of CE in antibody related studies, focussing especially on recent developments of CEIA technique. The principles for competitive and non-competitive CEIA are described with examples. Several detection methods and various applications are summarized and future developments in CEIA are speculated. CEIA has many potential applications, especially if the throughput is improved by using either multicapillary array or microchips with multiple channels. PMID- 9392389 TI - Determination of minute enzymatic activities by means of capillary electrophoretic techniques. AB - Capillary electrophoretic analysis of enzymes, co-enzymes, substrates and other chemical species that can be linked to an enzymatic reaction is reviewed with 80 references. Both off-line and on-line assays of minute enzymatic activities are discussed. In addition to heterogeneous on-line enzyme assays, a special emphasis is given to a newly established on-line technique called electrophoretically mediated microanalysis (EMMA). The basic principle, procedure, and various detection modes of EMMA are discussed. The recent developments in on-line determination of various enzyme substrates as well as on-line enzyme kinetic studies are also summarized. Some potential future developments in the determination of enzymatic activities by means of CE are also presented. PMID- 9392390 TI - Recent advances in chromatographic and electrophoretic methods for the study of drug-protein interactions. AB - Drug-protein binding is an important process in determining the activity and fate of a pharmaceutical agent once it has entered the body. This review examines various chromatographic and electrophoretic methods that have been developed to study such interactions. An overview of each technique is presented along with a discussion of its strengths, weaknesses and potential applications. Formats that are discussed include the use of both soluble and immobilized drugs or proteins, and approaches based on zonal elution, frontal analysis or vacancy peak measurements. Furthermore, examples are provided that illustrate the use of these methods in determining the overall extent of drug-protein binding, in examining the displacement of a drug by other agents and in measuring the equilibrium or rate constants for drug-protein interactions. Examples are also given demonstrating how the same methods, particularly when used in high-performance liquid chromatography or capillary electrophoresis systems, can be employed as rapid screening tools for investigating the binding of different forms of a chiral drug to a protein or the binding of different proteins and peptides to a given pharmaceutical agent. PMID- 9392391 TI - Glutaric aciduria type 1 (glutaryl-CoA-dehydrogenase deficiency): advances and unanswered questions. Report from an international meeting. AB - Infants with macrocephaly, young children with acute disease resembling encephalitis, and children with truncal hypotonia, ataxia, or dystonia may be affected by glutaric aciduria type I (GA 1, glutaryl-CoA-dehydrogenase deficiency), a not-so-rare autosomal recessive neurometabolic disease. Well-known features of GA1 are fronto-temporal brain atrophy with macrocephaly and acute encephalopathic episodes with striatal necrosis followed by dystonia, but some patients develop motor disease without overt crises and other biochemically affected individuals remain asymptomatic. Biochemical and molecular characterization is available and allows post- and prenatal diagnosis. The pathogenesis of fronto-temporal atrophy, macrocephaly, and basal ganglia necrosis is still not understood, and there is no close correlation between biochemical parameters and clinical outcome. There is, however, evidence suggesting that carnitine supplementation and anticatabolic treatment of intercurrent illness may arrest or prevent neurological deterioration, while the role of limitation of dietary lysine and tryptophane is not yet clear. Although pathogenetic aspects are poorly understood, the natural course of glutaric aciduria type 1 can be changed by early diagnosis and treatment. Coordinated research is needed to understand the pathogenesis of brain toxicity, to define the role of dietary therapy, and to explore the possibility of neonatal screening. PMID- 9392392 TI - Pituitary dwarfism in the R271W Pit-1 gene mutation. AB - The Pit-1 gene encodes the POU-domain transcription factor Pit-1 which is important for the differentiation of the anterior pituitary and regulation of the PRL, GH and TSH genes. As a member of the POU domain transcription factors, Pit-1 contains a DNA-binding region, consisting of a POU-specific domain and a POU homeodomain. Mutation of the Pit-1 gene causes hypoplasia of the pituitary gland and deficiencies of GH, PRL and TSH. In a DNA sample from a 3-month-old girl with severe growth deficiency from birth, single stranded conformational polymorphism analysis of the Pit-1 gene identified a gel shift in exon 6. DNA-sequencing disclosed a single base mutation in codon 271 (CGG to TGG) that changes arginine to tryptophan (R271W) in the POU homeodomain. The patient presented distinct facial features with prominent forehead, marked mid-facial hypoplasia with depressed nasal bridge, deep-set eyes and a short nose with anteverted nostrils. MRI examination showed a hypoplastic pituitary gland. Low serum GH did not respond to insulin-arginine provocation or GHRH tests. PRL levels below the detection limit did not increase in response to a TRH test. T4 and free T4 was below detection limit (< 20 nmol/l and < 4 pmol/l). TSH was 2.0 mU/l and showed a blunt response to 6.0 mU/l following TRH test. TBG was normal. In spite of inappropriately low TSH and very low T4, T3 was in the low normal range (1.4-1.6 nmol/l) and she was clinically euthyroid. The thyroid function tests are consistent with increased monodeiodination activity and increased conversion of T4 to T3, possibly related to the Pit-1 gene mutation. GH and T4 treatment resulted in catch-up growth continued during 5 years of therapy. CONCLUSION: Reports of nine other cases of R271W mutations of different populations as well as the present Norwegian patient suggest codon 271 of exon 6 to be a "hot spot" for Pit-1 mutations. To enable rapid and simple detection of this type of de novo mutation we have designed a specific amplification-created-restriction-site assay to check for the R271W mutation in patients suspected to have this rare form of genetic defect in growth hormone production. PMID- 9392393 TI - Combined pituitary deficiencies of growth hormone, thyroid stimulating hormone and prolactin due to Pit-1 gene mutation: a case report. AB - A child exhibited postnatal obstipation and icterus together with severe growth failure during the 1st year of life, a small facial skull and a prominent forehead. Endocrine work-up established the diagnosis of combined pituitary deficiencies of growth hormone, TSH and prolactin. Subsequently, the Pit-1 gene was analysed in the patient and both parents. A single point mutation was detected in exon 6 of the child: a C to G transversion on one allele, causing arginine in position 271 to be substituted by tryptophan (R271 W). This position is known as a "hot spot" for mutations. The inheritance is autosomal-dominant, as the mutated gene product interferes with DNA-binding of the wild-type protein. In contrast, other mutations in the PIT-1 gene are inherited in an autosomal recessive mode. CONCLUSION: Diagnosing Pit-1 gene mutations as a rare cause of combined pituitary deficiency is important both for genetic counselling as well as for predicting the future course in the patient (spontaneous puberty, no glucocorticoid substitution necessary during stress periods). PMID- 9392394 TI - Postprandial glycaemia after regular and lispro insulin in children and adolescents with diabetes. AB - We compared the postprandial blood glucose (BG)-levels following preprandial regular insulin or lispro insulin before and after eating in adolescents with diabetes. Lispro is a rapidly absorbed insulin analogue. Lispro insulin injected immediately before breakfast reduced the postprandial BG-rise significantly compared to the 20 min preprandially administered regular insulin (P < 0.01). Postprandial lispro injection resulted in similar BG values as the standard treatment with regular insulin 20 min preprandially. CONCLUSION: Lispro insulin injected immediately before the meal leads to lower postprandial BG levels and seems to be an option for teenagers who use multiple preprandial insulin injections. Postprandial lispro administration could be a benefit in certain situations since it resulted in similar BG values to preprandial regular insulin. PMID- 9392395 TI - Cure of infantile myofibromatosis with severe respiratory complications without antitumour therapy. AB - The prognosis of infantile myofibromatosis (IMF) depends on the organs involved: the prognosis is very poor if vital viscera are affected, but excellent if there is no visceral involvement. We report the case of a boy presenting with a pathological fracture at the age of 6 weeks. Progressive osteolytic lesions in the whole skeleton until the age of 8 months led to respiratory failure due to a softened thoracic wall requiring mechanical ventilation for 11 months. No pulmonary, laryngeal or other visceral involvement was found. In spite of the rapidly progressing disease and serious complications only supportive therapy was given. The lesions subsided gradually leaving slight deformities but normal function. At the age of 3.5 years the boy has an excellent quality of life. CONCLUSION: This case illustrates that even in progressing, complicated multifocal infantile myofibromatosis (without visceral involvement) the lesions can resolve without antitumour treatment if high quality intensive care is supplemented. PMID- 9392396 TI - Generalised bone disease with abundant periosteal reaction in megakaryocytic leukaemia. AB - We report an 18-month-old boy with trisomy 21 who presented with abundant, symmetrical periosteal hyperostosis and generalised osteolytic bone disease. Although adequate cytological and immunological studies have not been performed, the clinical course, routine blood and marrow studies allowed us to recognise megakaryoblastic leukaemia (ML) as the cause of these unique X-ray appearances. CONCLUSION: We present a unique case of generalised bone disease in an infant with trisomy 21. The appearances--clinical course and radiographic appearances- are consistent with ML. Such severe bony changes have not yet been reported in this association. This observation widens the spectrum of ML. PMID- 9392397 TI - Changes in coagulation and fibrinolysis in childhood acute lymphoblastic leukaemia re-induction therapy using three different asparaginase preparations. AB - Recently we reported the influence of two different Escherichia coli asparaginase (ASP) preparations on fibrinolytic proteins in childhood acute lymphoblastic leukaemia (ALL) demonstrating a significant association between ASP activity and haemostatic alterations. The present study was designed for prospective evaluation of coagulation and fibrinolytic parameters in leukaemic children receiving different ASP preparations during the course of re-induction. Forty leukaemic children receiving ASP (Medac: n = 13; Bayer: n = 10; Erwinia: n = 17) at 3-day intervals during re-induction were enrolled in this study. Blood samples for coagulation studies were obtained before each ASP administration together with serum samples for pharmacokinetic monitoring. Compared with Medac ASP 10,000 IU/m2, patients receiving Bayer ASP or Erwinia ASP showed significantly higher fibrinogen values. Antithrombin and plasminogen showed normal values in children after Erwinia ASP. Alpha2-antiplasmin and D-Dimer were no different in the groups studied. Neither side-effects, nor sustained asparagine depletion was observed in the majority of children treated with Erwinia ASP. CONCLUSION: Data of this study show a down-regulation of coagulation proteins in children treated with Medac ASP, less pronounced in patients after Bayer or Erwinia ASP. Since children treated with Erwinia ASP showed no adequate asparagine depletion during the course of ASP therapy, a dose adjustment should be discussed to guarantee asparagine depletion, the specific metabolic therapy for ALL. PMID- 9392398 TI - Conditions currently associated with erythema nodosum in Swiss children. AB - A review was made of the 36 paediatric patients in whom the diagnosis of erythema nodosum had been established between 1977 and 1996 at the Department of Paediatrics, University of Bern, Switzerland. Infectious diseases were associated with erythema nodosum in 20 (including 10 streptococcal infections) and non infectious inflammatory diseases in 8 patients. None of the 36 patients had tuberculosis or had been exposed to sulphonamides, phenytoin or hormonal contraceptives. There were eight patients in whom either the associated disease was not diagnosed, or there was no other disease. CONCLUSION: Most cases of erythema nodosum are nowadays caused by non-mycobacterial infectious diseases or by non-infectious inflammatory diseases. PMID- 9392399 TI - Vertical transmission of cytomegalovirus, most probably by breast milk, to an infant with Wiskott-Aldrich syndrome with fatal outcome. AB - A 4-month-old boy with prenatally diagnosed Wiskott-Aldrich syndrome became ill with a severe cytomegalovirus (CMV) infection, the outcome of which was fatal. The parents had isolated the infant from other children and adhered to standards of hygiene in order to avoid CMV infection because their first child had died of Wiskott-Aldrich syndrome and CMV infection. The mother breast-fed her child although she was CMV IgG positive. The source of infection was most probably breast milk, which contained CMV at the time the infant developed the generalized CMV infection. CONCLUSION: In infants with immunodeficiency syndromes, CMV infection may have a fatal outcome. Since the virus can be transmitted by breast milk, the advantages and disadvantages of breast-feeding should, therefore, be weighed in newborn infants with an immunodeficiency syndrome whose mother is a CMV carrier. PMID- 9392400 TI - Septic arthritis of the hip by Fusobacterium necrophorum after tonsillectomy: a form of Lemierre syndrome? AB - Lemierre syndrome used to be a complication of severe oropharyngeal infection with regional thrombophlebitis, septicaemia and septic metastatic infections caused by Fusobacterium necrophorum in the pre-antibiotic era. A case of septic arthritis of the hip caused by F. necrophorum as a complication of tonsillectomy is reported in a 9-year-old boy. CONCLUSION: Lemierre syndrome, usually seen after an oropharyngeal infection, can also complicate tonsillectomy. PMID- 9392401 TI - Intramuscular ceftriaxone compared with oral amoxicillin-clavulanate for treatment of acute otitis media in children. AB - Two hundred and fifteen children aged 4 months 6 years with acute otitis media (AOM) were randomized to be treated either by a single i.m. injection of ceftriaxone, 50 mg/kg, with a second dose in the event of unsatisfactory response after 48 h or a history of recurrent AOM (109 patients) or amoxicillin clavulanate 12.5 mg tid (106 patients). The failure rate was similar in children treated by ceftriaxone and amoxicillin clavulanate, 4.6% and 4.7%, respectively (standard error for intergroup difference -2.87%, 95% confidence interval -5.62% to 5.87%). No significant differences between the groups were found in the dynamics of the resolution of the acute symptomatology, otoscopy findings, relapse rate at 30 days or tympanographic evidence of middle ear effusion at the scheduled visits on days 30, 60 and 90. Recurrence of AOM between days 31 and 90 was observed significantly in more children treated with amoxicillin clavulanate than with ceftriaxone--25 out of 84 (29.4%) versus 11 out of 81 (13.6%) (P = 0.012). CONCLUSION: Ceftriaxone injection(s) is as efficient at least as 10-day oral amoxicillin clavulanate for treatment of acute otitis media in children. Although not recommended as routine, ceftriaxone can be considered in the management of acute otitis media under special circumstances, particularly in cases when the ability to tolerate or absorb oral drugs is compromised, in children refusing or unable to take oral therapy or when the compliance is questionable. PMID- 9392402 TI - Acquired carnitine abnormalities in critically ill children. AB - In order to characterize the role of carnitine during metabolic stress, we prospectively determined carnitine profiles in plasma and urine on admission, days 2, 5, 10 and 15, among 28 critically ill children free of any known conditions associated with secondary carnitine deficiency. More than 25% of plasma and 50% of urinary carnitine measurements were abnormal; 96% (27/28) of patients displayed on at least one occasion an abnormal [< -2 SD or > +2 SD] carnitine value in plasma. Three children had extremely low [< 10 micromol/l] free carnitine (FC) levels in plasma. Plasma esterified and FC levels on admission were not related to the risk of mortality [PRISM score], to muscle lysis [CK values], and to the caloric intake. Levels of FC and esterified carnitine in plasma were unrelated to those measured in urine. CONCLUSION: Abnormal plasma and urine carnitine measurements are frequently found in critically ill children; the biological significance of these perturbations remains unclear. Caution must be exercised before concluding that an abnormal carnitine value is indicative of an underlying hereditary metabolic disorder in this population. PMID- 9392403 TI - Succinyl-CoA:acetoacetate transferase deficiency: identification of a new patient with a neonatal onset and review of the literature. AB - We describe the clinical symptoms and biochemical findings of a patient with succinyl-CoA:acetoacetate transferase deficiency who presented in the neonatal period and review the current literature on this subject. Our patient was initially suspected to have distal renal tubular acidosis, and subsequently, a fasting test revealed severe metabolic ketoacidosis with normal blood glucose after 13 h which suggest a defect in ketolysis. In his cultured skin fibroblasts succinyl-CoA:acetoacetate transferase was deficient (residual activity 15%). Treatment in the acute phase consisted of sodium bicarbonate. At the present age of 9 years, psychomotor and physical development are within normal limits. CONCLUSION: Defects of ketolysis probably are underdiagnosed disorders and should be considered in infants and young children with persistent ketosis. PMID- 9392404 TI - UNICEF/WHO baby-friendly hospital initiative: does the use of bottles and pacifiers in the neonatal nursery prevent successful breastfeeding? Neonatal Study Group. AB - To promote breastfeeding, UNICEF/WHO have launched the "baby-friendly hospital initiative" focusing on hospital care routines during delivery and the first days of life. In industrialised countries, two aspects of the initiative have raised controversy: how do restriction of supplemental feedings and ban of bottles and pacifiers affect long-term breastfeeding performance? From ten centres 602 healthy newborns were randomly assigned either to a UNICEF group with restrictive fluid supplements and avoidance of bottles and pacifiers during the first 5 days of life, or to a standard group with conventional feeding practice. Breastfeeding was encouraged in both groups. The main study endpoints were the prevalences of breast-feeding on day 5, and after 2, 4 and 6 months. Of the newborns 46% violated the UNICEF protocol, mostly because of maternal requests to give a pacifier or supplements by bottle. In the standard group, the drop-out rate was 9.7%. No significant differences in breastfeeding frequency and duration could be found: (UNICEF vs standard) day 5: 100% vs 99%; 2 months: 88% vs 88%; 4 months: 75% vs 71%; 6 months: 57% vs 55%. Inclusion of drop-outs due to pacifier use did not alter the results. CONCLUSION: In our study population fluid supplements offered by bottle with or without the use of pacifiers during the first 5 days of life were not associated with a lower frequency or shorter duration of breastfeeding during the first 6 months of life. PMID- 9392405 TI - Home oxygen therapy in infants with bronchopulmonary dysplasia: a prospective study. AB - We followed the clinical course of 21 infants with bronchopulmonary dysplasia enrolled in a prospective home O2 therapy programme during a 4-year-period. Mean gestational age was 28.5 weeks (range, 25-36 weeks) and mean birth weight 1093 g (range 630-2750 g). Infants were regularly monitored to maintain pulse oximeter O2 saturation over 94%-95%. The source of O2 was liquid oxygen and was delivered by nasal cannula. During the follow up oxygenation was assessed by SatO2 measurement, cardiac function by Doppler echocardiography and respiratory function by the occlusion technique. All patients had an ophthalmological follow up. The mean age of the infants at discharge was 3.7 months (range 1.7-8.6) and mean weight 2830 g (range 2150-3780 g). At discharge 8 infants had right ventricular hypertrophy (RVH) and four of them had pulmonary hypertension. Mean duration of home O2 therapy was 97 days (range 15-320 days) and the mean age of discontinuation of O2 was 6.9 months (range 3-14.7 months). The cardiological follow up was benign: the ECG signs of RVH disappeared by 12 months of age in six out of eight infants and the right ventricular pulmonary pressure, as measured by the Doppler method, normalised in the four patients in whom it was detected. No relationship was found between respiratory mechanics and the duration of O2 therapy. Weight gain was poor with mean growth at the 3rd percentile for females and just below the 3rd percentile for males. Twelve of the 21 infants required 25 rehospitalizations. No one presented deterioration of retinopathy of prematurity that was present in 16 infants at discharge; at 12 months retinopathy was resolved in 14 infants. A total of 2025 hospital days were saved, representing a significant financial saving. CONCLUSION: Home O2 therapy permits the safe early discharge of O2-dependent BPD infants and it reduces significantly the length of time spent in hospital which represents a considerable financial saving. PMID- 9392407 TI - A girl with congenital adrenal hyperplasia and recurrent abdominal pain. PMID- 9392406 TI - Altered cardiovascular autonomic regulation after 2-week inhaled salbutamol treatment in asthmatic children. AB - We studied the effects of therapeutic 2-week inhaled salbutamol treatment on the cardiovascular and respiratory autonomic nervous regulation in eight children with asthma. In this randomized, double-blind, placebo-controlled crossover study our test subjects inhaled 200 microg salbutamol or placebo thrice daily for 14 days. After the 14-day treatment we continuously measured electrocardiogram, finger systolic arterial pressure (SAP) and flow-volume spirometry at baseline and the response to a single 600 microg salbutamol inhalation. The periodic variability components of R-R intervals (the time between successive heart beats) and SAP in relation to respiration were assessed using spectral analysis. Two week salbutamol treatment increased baseline low frequency (LF) variability (P < 0.05) and low frequency/high frequency (LF/HF) variability ratio of R-R intervals (P < 0.05) when compared to the placebo treatment. As a response to the single salbutamol inhalation the increase in LF/HF ratio of R-R intervals was smaller after the 2-week salbutamol treatment (P < 0.01). No significant differences were found in the bronchodilatory response after the treatment period. CONCLUSION: Two week salbutamol treatment shifts the cardiovascular autonomic regulation to a new level characterized by greater sympathetic responsiveness and slight beta2 receptor tolerance. Because these effects were evident 18 h after cessation of the therapy they are likely to reflect the adaptation of organ responses to regular therapy or altered central autonomic regulation rather than direct drug effect. A slight tolerance developed in the sympathovagal cardiac response but not in the bronchodilatory response. PMID- 9392408 TI - A neonate with respiratory distress after a traumatic delivery. PMID- 9392409 TI - Visualization of injection depot after subcutaneous administration by syringe and needle-free device (Medi-Jector): first results with magnetic resonance imaging. PMID- 9392410 TI - Lack of insulin-like growth factor I for therapeutic research. European Society for Paediatric Endocrinology. PMID- 9392411 TI - Dangers of recreational helmet use in playgrounds. PMID- 9392412 TI - Juvenile pityriasis rubra pilaris with isolated IgA deficiency. PMID- 9392413 TI - Balanced translocation t(4q; 10q) in infantile spinal muscular atrophy. PMID- 9392414 TI - Transumbilical venous access with small diameter silastic catheters in very low birth weight infants. PMID- 9392415 TI - Macroscopic haematuria following immunisation with tetanus toxoid and oral polio vaccine. PMID- 9392416 TI - Hepatic uptake of chylomicron remnants. AB - Chylomicrons are formed in the intestine and transport dietary triglyceride to peripheral tissues and cholesterol to the liver. The enzyme lipoprotein lipase, with apolipoprotein (apo)C-II as a co-factor, hydrolyzes chylomicron triglyceride allowing the delivery of free fatty acids to muscle and adipose tissue. As a result, a new particle called a chylomicron remnant is formed. This particle is enriched in cholesteryl ester and fat-soluble vitamins and contains apoB-48 and apoE. It is rapidly removed from the circulation by the liver. ApoE is the moiety required for rapid hepatic removal. Its activity is inhibited by C apolipoproteins, especially apoC-I. Hepatic removal appears to be accomplished by several overlapping mechanisms. The particle must first achieve a size that allows it to be "sieved" through the endothelial fenestre allowing entrance into the space of Disse. Here, it may 1) be removed directly by LDL receptors; 2) acquire additional apoE that is secreted free into the space, and then be removed directly by the LDL receptor-related protein (LRP); or 3) it may be sequestered in the space. Sequestration occurs by binding of apoE to heparan sulfate proteoglycans and/or binding of apoB to hepatic lipase. Sequestered particles may be further metabolized allowing apoE, and lysophospholipid enrichment, followed by transfer to one of the above receptors for hepatic uptake. The above formulation is based upon animal studies. In humans, delayed removal of chylomicron remnants has been documented in diabetes, renal failure, and familial combined hyperlipemia and is the abnormality resulting in type III hyperlipidemia. Case control studies have identified delayed remnant removal as an independent risk factor for atherosclerotic cardiovascular disease. Thus, understanding the further details of the processes, and how it can be regulated in humans, is an important challenge for the future. PMID- 9392417 TI - Learning about the structure and biology of human lipoprotein [a] through dissection by enzymes of the elastase family: facts and speculations. AB - Lipoprotein[a], Lp[a], represents a class of lipoprotein particles that have as a protein moiety apoB-100 linked by a disulfide bridge to a multi-kringle structure, apolipoprotein[a], or apo[a]. It is now possible to separate from Lp[a] a free apo[a] able to reassociate with apoB-100-containing lipoproteins to restore the parent lipoprotein complex. Apo[a], whether free or a constitutive component of Lp[a], can be cleaved at interkringle sites by the action of enzymes of the elastase family generating fragments that differ in structural, functional, and metabolic properties. In the case of Lp[a], elastase digestion generates a miniLp[a] particle, which contains the apo[a] COOH-terminal domain able to bind to lysine, fibrinogen, fibronectin, and proteoglycans. This domain may also be generated by elastase type enzymes secreted by activated macrophages and smooth muscle cells in the arterial intima as a part of the chronic inflammation that characterizes the atherosclerotic process. Thus, the apo[a] immunoreactive material, which has been described in the atherosclerotic plaque, may represent miniLp[a] and/or apo[a] fragments accumulating in the vessel wall as a function of their relative affinity for the components of the extracellular matrix and producing complexes with an atherothrombogenic potential. This potential may depend on several factors: kringle folding and conformation, susceptibility of the linkers to proteolytic cleavage, binding specificity of given apo[a] fragments to the matrix components of the arterial intima, and the overall inflammatory status of the arterial wall. PMID- 9392418 TI - Fresh mouse peritoneal macrophages have low scavenger receptor activity. AB - Peritoneal macrophages are easily isolated by lavage, suggesting that they are either nonadherent or weakly adherent in situ. Cultured macrophages express class A scavenger receptors (SCR), which mediate Ca2+-independent adhesion in vitro. We examined fresh peritoneal macrophages from mice and from women with endometriosis to determine whether the adherence of these cells was associated with increased expression of class A SCR. Fresh human macrophages were not immunoreactive to SCR antibodies; however, SCR immunoreactivity increased with time in culture. Fresh mouse and human macrophages took up minimal amounts of 1,1'-dioctadecyl-3,3,3',3' tetramethylindocarbocyanine (DiI)-acetyl-low density lipoproteins (Ac-LDL), a class A SCR ligand. Murine macrophages in culture for 24-72 h internalized four times more Ac-LDL than fresh cells. Cells cultured for 2 days incorporated 3.2 times more [14C] oleate than freshly isolated cells (55.7 +/- 7.9 versus 17.6 +/- 3.0 nmol/mg cell protein). In contrast to SCR activity, mouse macrophage SCR mRNA expression was similar in freshly isolated macrophages and those cultured for 3 days. These results suggest that peritoneal macrophages express only low levels of SCR activity in situ and that posttranscriptional regulation after isolation leads to an increase in SCR activity that correlates with adherence of the macrophages in vitro. PMID- 9392419 TI - Post-translational regulation of mevalonate kinase by intermediates of the cholesterol and nonsterol isoprene biosynthetic pathways. AB - To assess the potential for feedback inhibition by isoprene intermediates in the cholesterol and nonsterol isoprene biosynthetic pathway, we expressed human cDNAs encoding mevalonate kinase (MKase), phosphomevalonate kinase (PMKase), and mevalonate diphosphate decarboxylase (MDDase) as fusion proteins in Escherichia coli DH5alpha, and purified these proteins by affinity chromatography. Several phosphorylated and non-phosphorylated isoprenes were analyzed as inhibitors of the enzymes using a standard spectrophotometric assay. Of the three proteins, only MKase was inhibited through competitive interaction at the ATP-binding site. The intermediates studied (and their relative inhibitory capacity) were: geranylgeranyl-diphosphate (GGPP, C20) > farnesyl-diphosphate (FPP, C15) > geranyl-diphosphate (GPP, C10) > isopentenyl-diphosphate (IPP, C5) > or = 3,3 dimethylallyl-diphosphate (DMAPP, C5) > farnesol (C15) > dolichol-phosphate (DP, C(80-100)). Mevalonate-diphosphate, geraniol, and dolichol were not inhibitors. Our data further define the spectrum of physiologic inhibitors of MKase, and provide the first evidence for feedback inhibition of MKase by a nonsterol isoprene produced by the branched pathway, dolichol-phosphate. These results provide additional evidence that MKase may occupy a central regulatory role in the control of cholesterol and nonsterol isoprene biosynthesis. PMID- 9392420 TI - Diacylglycerol is the preferred substrate in high density lipoproteins for human hepatic lipase. AB - The hydrolysis of HDL phospholipids (PL) and glycerides by hepatic lipase (HL) has been investigated in native and reconstituted HDL particles (Lp2A-I). Fasting, normolipidemic HDL exhibit total lipid hydrolytic rates of between 10 and 36 nM FA/h per microM PL. Of the total fatty acids liberated with HDL3 only 1% are from triolein (TG), while 49% are from diolein (DG) and 50% are from PL. A spherical reconstituted particle containing 2 molecules of apoA-I, 120 molecules of PL, and 20 molecules of TG exhibits a total lipid hydrolytic rate of 18 nM FA/h per microM PL and 93% of the fatty acids liberated are from PL. Inclusion of 40 molecules of TG into the Lp2A-I particle doubles the rate of fatty acid hydrolysis by HL through a stimulation of TG hydrolysis. Further addition of 10 molecules of DG to the Lp2A-I complex has no effect on the overall rates of hydrolysis, but changes the substrate specificity, wherein 61% of the fatty acids are from DG and both TG and PL hydrolytic rates are significantly reduced. Increasing the amount of DG in the Lp2A-I particle further stimulates total lipid hydrolysis by raising DG hydrolytic rates at the expense of PL and TG hydrolysis. A particle containing 10 molecules of TG and 40 molecules DG yields the fastest lipid hydrolytic rate of 143 nM FA/h per microM PL, which constitutes 96% DG hydrolysis, 3% TG hydrolysis, and 1% PC hydrolysis. These data indicate that hepatic lipase acts primarily as a surface lipid lipase with HDL particles. DG is the preferred substrate of HL in HDL and the HDL-DG content regulates the hydrolysis of both PL and TG by HL. PMID- 9392421 TI - Secretory non-pancreatic phospholipase A2: influence on lipoprotein metabolism. AB - Lipoprotein metabolism is markedly altered during inflammation. The concentration of human secretory phospholipase A2 (sPLA2) can increase hundreds of fold in inflammatory fluids and in the circulation. It was detected in atherosclerotic lesions where many inflammatory genes are induced. As sPLA2 has been reported to act on lipoproteins as substrates, lipoprotein profiles in transgenic mice expressing sPLA2 were studied. HDL levels were markedly decreased in transgenic mice overexpressing sPLA2. HDL in the transgenics were smaller in size, with a significant decrease (11%) in phospholipid content compared to nontransgenic littermates. In sPLA2 transgenic mice and transgenic mice expressing both sPLA2 and human apolipoprotein B (apoB), the concentrations of apoB-containing lipoproteins were not altered. We conclude that sPLA2 alters HDL metabolism and could be responsible for the depressed levels of HDL that exist during chronic inflammatory diseases. PMID- 9392422 TI - Phorbol ester-induced low density lipoprotein receptor gene expression in HepG2 cells involves protein kinase C-mediated p42/44 MAP kinase activation. AB - The signaling pathway involved in low density lipoprotein (LDL) receptor gene expression induced by the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) was investigated in the human hepatoma HepG2 cell line. Treatment of HepG2 cells with 100 nM TPA resulted in an approximately 20-fold increase in LDL receptor mRNA level, as determined by RT-PCR, which peaked at 2-4 h of treatment and subsequently declined. The protein kinase C (PKC) inhibitors calphostin C and staurosporine prevented TPA-mediated LDL receptor mRNA induction. In contrast, TPA did not affect squalene synthase mRNA expression. Immunoblotting of cell extracts with isozyme-specific PKC antibodies revealed that HepG2 cells expressed PKC alpha, which was mainly cytosolic, and PKC beta, PK epsilon, and PKC zeta, all of which were present in both the cytosolic and particulate fractions. Treatment of HepG2 cells with 100 nM TPA resulted in translocation of cytosolic PKC alpha to the particulate fraction, with a maximum at 30 min-2 h of treatment, but was without effect on the subcellular distribution of the other isozymes. TPA treatment also led to activation of the mitogen-activated protein kinase (MAPK) ERK cascade. The specific MAPK pathway inhibitor PD98059 blocked TPA-induced ERK activation. Furthermore, pretreatment of cells with PD98059 inhibited TPA-induced LDL receptor mRNA induction. Moreover, pretreatment of cells with calphostin C inhibited TPA-mediated ERK activation and LDL receptor mRNA induction in a dose dependent fashion. Based on a close kinetic correlation between PKC alpha translocation and ERK activation, and the effects of specific inhibitors, these findings suggest that translocation/activation of PKC alpha, and subsequent activation of the Raf-1/MEK/ERK MAPK cascade, represent key events in the transcriptional induction of LDL receptor gene by TPA in HepG2 cells. PMID- 9392423 TI - Adipose differentiation-related protein is an ubiquitously expressed lipid storage droplet-associated protein. AB - The adipose differentiation-related protein (ADRP) was first characterized as a mRNA induced early during adipocyte differentiation (Jiang, H. P., and G. Serrero. 1992. Proc. Natl. Acad. Sci. USA. 89:7856-7860). The present study demonstrates that ADRP mRNA is expressed in a variety of tissues and cultured cell lines. Immunocytochemical examination revealed that ADRP localizes to neutral lipid storage droplets in cultured murine 3T3-L1 adipocytes, murine MA-10 Leydig cells, Chinese hamster ovary (CHO) fibroblasts, and human HepG2 hepatoma cells; the association of ADRP with lipid droplets was confirmed by subcellular fractionation of MA-10 Leydig cells. In addition to ADRP, steroidogenic cells and adipocytes express the perilipins, a family of lipid droplet-associated proteins that share a highly related sequence domain with ADRP. ADRP and perilipins co localize on lipid droplets in MA-10 Leydig cells. While ADRP was found on small lipid droplets in 3T3-L1 preadipocytes and early differentiated adipocytes, it was absent in maturing adipocytes. In contrast, perilipins were absent early during differentiation, but were found on small and large lipid droplets at later stages. The transition in surface protein composition of adipocyte lipid droplets from ADRP to perilipins occurred 3 days after the initiation of differentiation when cells displayed co-localizatioin of both proteins on the same lipid droplets. The specific localization of adipose differentiation-related protein to lipid droplets in a wide variety of cells suggests that ADRP plays a role in management of neutral lipid stores. PMID- 9392424 TI - Use of cyclodextrins for manipulating cellular cholesterol content. AB - Previous studies from this laboratory have demonstrated that exposure of tissue culture cells to cyclodextrins results in rapid cholesterol depletion. In the present study, we have developed experimental systems for using solutions of cyclodextrins, either 2-hydroxypropyl beta-cyclodextrin or methylated beta cyclodextrin, complexed with varying amounts of free cholesterol to manipulate cell cholesterol content. Cholesterol delivered via the cyclodextrin has been found to be metabolically active, as measured by the acyl-coenzyme A:cholesterol acyltransferase (ACAT)-mediated esterification of [3H]cholesterol in Fu5AH rat hepatoma cells and Chinese hamster ovary cells. The methylated beta-cyclodextrin was found to be a more efficient donor in all cell types studied, with an average cholesterol uptake of at least 100 microg cholesterol/mg protein within 6 h. By modifying the cyclodextrin:cholesterol molar ratio, it is possible to manipulate the cellular cholesterol content of cells, producing conditions ranging from net cholesterol enrichment to depletion. The use of cyclodextrins provides a convenient, precise and reproducible method for modulating the cholesterol content of tissue culture cells. PMID- 9392425 TI - Mechanism of the slow induction of apolipoprotein A-I synthesis by retinoids in cynomolgus hepatocytes: involvement of retinoic acid and retinoid X receptors. AB - We showed previously that retinoids stimulate apolipoprotein A-I (apoA-I) synthesis in cultured cynomolgus hepatocytes only after a 24-h lag phase. Here we report on the biochemical background of the slow response, the requirement for high retinoic acid concentrations, and the involvement of different retinoid receptors. The time course of the effect of 10 microM all-trans retinoic acid (at RA) on apoA-I mRNA levels and protein secretion were comparable, i.e., minor increases were observed after a 24-h incubation and mRNA levels were increased 2.2- and 3.5-fold after 48 h and 72 h, respectively. In contrast, apoA-I gene transcription was already increased (2.6-fold) after a 4-h incubation with 10 microM at-RA. At-RA disappeared rapidly from the cultures: after 2 h of incubation 40% of the added amount was left and after 24 h only 2%. RAR beta mRNA and gene expression were increased after incubation with 10 microM at-RA, whereas RAR alpha and RXR alpha mRNA levels and expression remained unchanged. No transcriptional activity and mRNA for other retinoid receptors were detectable. Both RAR-selective (TTNPB) and RXR-selective (3-methyl-TTNEB) agonists induced apoA-I synthesis at 1 and 10 microM. These results show that i) the slow increase in apoA-I secretion is caused by a slow increase of its mRNA level; ii) the apoA I gene transcription in cynomolgus hepatocytes is induced rapidly by retinoids; iii) the added at-RA disappeared rapidly from the cultures, explaining the necessity for high initial concentrations; iv) RLR alpha and/or RAR beta and RXR alpha are involved in the activation of apoA-I expression by retinoids. PMID- 9392426 TI - Determinants of plasma platelet-activating factor acetylhydrolase: heritability and relationship to plasma lipoproteins. AB - Plasma platelet-activating factor acetylhydrolase (PAF-AH) is the enzyme that inactivates PAF (1-alkyl-2-acetyl-sn-glycero-3-phosphocholine). We determined the relative contributions of genetic and environmental factors to variation in plasma PAF-AH activity in 240 individuals from 60 nuclear families. Regression of mean-offspring PAF-AH activity on the mid-parent value indicated that 62% of the variation in plasma PAF-AH activity was heritable. Spousal values were weakly negatively correlated, indicating that familial aggregation of PAF-AH activity is due to genetic rather than to environmental factors. Among normolipidemic individuals, plasma PAF-AH activity was strongly correlated with the plasma concentration of low density lipoprotein cholesterol (LDL-C), and treatment with lovastatin resulted in proportional decreases in plasma PAF-AH activity and LDL-C concentrations. To further elucidate the relationship between PAF-AH and plasma concentrations of LDL, plasma PAF-AH activity was measured in families with well defined, monogenic disorders of LDL metabolism. Plasma PAF-AH activity cosegregated with plasma LDL-C concentrations in familial hypercholesterolemia, but not in familial hypobetalipoproteinemia. We speculate that the rate of removal of LDL from the circulation may determine the clearance rate of PAF-AH, thereby modulating the activity of PAF-AH in blood. PMID- 9392427 TI - Diet modification alters plasma HDL cholesterol concentrations but not the transport of HDL cholesteryl esters to the liver in the hamster. AB - These studies were undertaken to investigate the mechanism whereby diet modification alters the plasma concentration of high density lipoprotein (HDL) cholesteryl ester and apoA-I and to determine whether diet-induced alterations in circulating HDL levels are associated with changes in the rate of reverse cholesterol transport. Rates of HDL cholesteryl ester and apoA-I transport were measured in hamsters fed a control low-cholesterol, low-fat diet or the same diet supplemented with soluble fiber (psyllium) or with cholesterol and triglyceride (Western-type diet). The Western-type diet increased the plasma concentration of HDL cholesteryl ester by 46% compared to the control diet and by 86% compared to the psyllium-supplemented diet; nevertheless, the absolute rates of HDL cholesteryl ester transport to the liver were identical in the three groups. Diet induced alterations in circulating HDL cholesteryl ester levels were due to changes in the rate of HDL cholesteryl ester entry into HDL (whole body HDL cholesteryl ester transport) and not to regulation of HDL cholesteryl ester clearance mechanisms. The Western-type diet increased the plasma concentration of HDL apoA-I by 25% compared to the control diet and by 45% relative to the psyllium-supplemented diet. Diet-induced alterations in plasma HDL apoA-I concentrations were also due entirely to changes in the rate of apoA-I entry into HDL (whole body HDL apoA-I transport). These studies demonstrate that the absolute flux of HDL cholesteryl ester to the liver, which reflects the rate of reverse cholesterol transport, remains constant under conditions in which plasma HDL cholesteryl ester concentrations are altered over a nearly 2-fold range by diet modification. PMID- 9392428 TI - A two-base deletion in exon 6 of the 3-hydroxy-3-methylglutaryl coenzyme A lyase (HL) gene producing the skipping of exons 5 and 6 determines 3-hydroxy-3 methylglutaric aciduria. AB - A novel two-base deletion in the 3-hydroxy-3-methylglutaryl coenzyme A lyase (HL) gene was found in a Spanish patient with homozygous 3-hydroxy-3-methylglutaric aciduria. Amplification by RT-PCR of the mRNAs showed that the gene was transcribed into three different mRNAs. One showed the complete deletion of exons 5 and 6 located between nucleotides 348 and 561 of the HL cDNA. The second transcript showed deletion of exon 6 only, and the third contained a two-base deletion CT in exon 6, corresponding to nucleotides 504 and 505 of the HL cDNA. These aberrant mRNAs are predicted to encode three abnormal HMG-CoA lyase proteins; the first (from skipped exons 5 and 6) lacks 71 amino acids, which represents 24% of the mature protein; the second, (from the skipping of exon 6, producing a frameshift) contains only 192 amino acids, the last 26 of which are missense amino acids preceding a stop codon; the third contains only 175 amino acids, the last 7 of which are missense. Northern blot analysis showed that the HL mRNA levels of the patient were 4% of the control. PCR quantitative analysis indicated that the mRNA lacking exons 5 and 6 was the most abundant, representing 88% of the total. The other two mRNAs represented 8% and 4%, respectively. In the genomic DNA only one CT deletion was found at positions +7 and +8 at beginning of exon 6. No mutations were observed in the splice donor, splice acceptor, or pyrimidine-rich sequences of the intronic regions flanking exons 5 and 6. All three aberrant mRNAs resulted only from the deletion of nucleotides CT. We suggest that this deletion may affect the interaction between the small nuclear ribonucleoproteins (snRNPs) and exon 6, and that, as a result, the abnormal splicing of the pre-mRNA produces two different aberrant transcripts. PMID- 9392430 TI - Four novel mutations of sterol 27-hydroxylase gene in Italian patients with cerebrotendinous xanthomatosis. AB - We report the characterization of eight mutations of sterol 27-hydroxylase gene (CYP27) in five Italian patients with cerebrotendinous xanthomatosis, who were found to be compound heterozygotes. Four mutations (C --> T at nt 45 of exon 4, G(+1) --> A in intron 6, G(+5) --> T in intron 7, and G(-1) --> A in intron 7) are novel. The C --> T at nt 45 of exon 4 converts the arginine codon into a stop codon thus generating a truncated protein of 198 amino acids. The three splice site mutations reduced the content of CYP27 mRNA in skin fibroblasts to very low or undetectable levels and generated minute amounts of abnormal mRNAs. The G(+1) -> A transition in intron 6 produced three abnormal mRNAs. In the first, the 5' half of exon 6 joins to exon 7, skipping 89 bp of exon 6, and in the second, exon 5 joins directly to exon 7. The predicted translation products of these mRNAs are truncated proteins. In the third abnormal mRNA, exon 5 joins to exon 8 with an in frame deletion of 246 bp. The G(+5) --> T transversion in intron 7 generates a single abnormal mRNA in which exon 6 joins directly to exon 8, with a frameshift and a premature stop codon. In the G(-1) --> A transition in intron 7, two mRNAs are generated. In the first, the retention of the whole intron 7 causes a frameshift and a premature stop codon; in the second, the joining of exon 7 to exon 8 is associated with an in-frame deletion of the first 6 nucleotides. All these novel mutations are predicted to produce structurally abnormal enzymatic proteins with no measurable biological activity. PMID- 9392429 TI - High density lipoprotein particle size restriction in apolipoprotein A-I(Milano) transgenic mice. AB - Human carriers of apolipoprotein A-I(Milano) (Arg173 --> Cys substitution in apolipoprotein A-I) are characterized by an HDL deficiency in which small, dense HDL accumulate in plasma. Because affected individuals are heterozygous for this mutation, the full impact of apolipoprotein A-I(Milano) (apoA-I(Milano)) on HDL cholesterol metabolism is unknown. In this study, apoA-I(Milano) transgenic mice were used to evaluate the extent of apoA-I(Milano) dimerization and HDL particle size restriction in the absence of wild-type apoA-I. Murine apoA-I knockout mice were utilized to express apoA-I(Milano) and human apoA-II in the presence of wild type, human apoA-I (apoA-IMilano/A-Iwt/A-II) and in its absence (apoA-IMilano/A II). Plasma HDL-cholesterol concentrations were similar (30 mg/dl) in both lines of apoA-I(Milano) transgenic mice. In the apoA-IMilano/A-Iwt/A-II phenotype, 14% of the apoA-I(Milano) formed homodimers and 33% formed heterodimers with apoA-II. ApoA-I(Milano) homodimers increased by 71% in the apoA-IMilano/A-II transgenics and was associated with an abundance of small, 7.6-nm HDL3-sized particles compared to the 9.5, 8.3, and 7.6-nm-sized particles in apoA-IMilano/A-Iwt/A-II mice. The unesterified cholesterol/cholesteryl ester mole ratio of HDL was elevated by 45% in apoA-IMilano/A-Iwt/A-II mice and by 90% in apoA-IMilano/A-II transgenics compared to wild-type (human apoA-I/A-II). Both apoA-I(Milano) transgenics possessed normal levels of plasma LCAT activity, but endogenous cholesterol esterification rates were reduced by 50% compared to controls. Thus, HDL particle size restriction was not the result of impaired LCAT activation; rather, dimerization of apoA-I(Milano) limited the esterification of cholesterol on endogenous HDL. In the absence of wild-type apoA-I, the more extensive dimerization of apoA-I(Milano) severely limited cholesteryl ester accumulation on plasma HDL accounting for the abundance of small, 7.6-nm HDL3 particles in apoA IMilano/A-II mice. PMID- 9392431 TI - Clearance of lipoprotein remnant particles in adipose tissue and muscle in humans. AB - A major proportion of triglycerides in plasma triglyceride-rich lipoproteins (TRL) are removed in peripheral tissues by lipoprotein lipase, and hypothetically a minor proportion can also be removed by whole-lipoprotein particle uptake. This second removal pathway has not previously been directly demonstrated in humans. Simultaneous blood samples were drawn from arterialized blood, a vein draining the subcutaneous abdominal adipose tissue, and a deep antecubital vein of the forearm to provide arterio-venous gradients from blood-draining adipose tissue and muscle in seven male subjects. The men were given a fat-rich mixed meal containing vitamin A and the triglyceride and retinyl palmitate (RP) concentrations were quantified in the plasma. Density gradient ultracentrifugation was used to isolate TRL fractions, in which triglycerides, RP, apoB-48, and apoB-100 were quantified. There was clearance of triglycerides in muscle and adipose tissue and, in addition, removal of RP. By analysis of the TRL subfractions, the RP removal was likely to be confined to the largest chylomicron remnant particles. For the Sf > 400 fraction, the area under curve (AUC) relative to arterial for triglycerides were 79% (66-91%) and 81% (72-89%) in adipose tissue and muscle venous outflow, respectively (each P < 0.02 versus arterial). The corresponding values for RP were 87% (73-101%) and 85% (69-100%), respectively, (each P < 0.05 versus arterial). In the Sf 60-400 fraction there was further uptake of triglycerides, but not of RP. We hypothesize that the periphery could be of importance for removal of the largest chylomicron remnants, as their size might partially exclude them penetrating the fenestrated hepatic sinusoidal endothelium to reach the hepatic chylomicron remnant receptors. PMID- 9392432 TI - Transforming growth factor beta is sequestered into an inactive pool by lipoproteins. AB - Elevated plasma concentrations of low density lipoprotein (LDL) and very-low density lipoprotein (VLDL) have been correlated with the development of atherosclerosis. These lipoproteins may promote atherogenesis by direct deposition of lipid in the vessel wall. In addition, previous data suggested that there was an inverse correlation between serum LDL-cholesterol concentration and the proportion of transforming growth factor beta (TGF-beta) in an active form (Grainger et al. 1995. Nature Med. 1:74). Here we have investigated whether lipoproteins can affect the activity of TGF-beta1 in plasma and show that TGF beta can associate with the lipoprotein fraction. In the plasma of healthy males, 16 +/- 5% (mean +/- standard deviation; n = 57) of the total plasma TGF-beta1 was associated with the lipoprotein fraction, with the major proportion (64 +/- 15%) in the HDL-3 subfraction. However, in ten diabetic subjects with moderately poor glucose control (Hb alc > 8.0), the proportion of total plasma TGF-beta in the lipoprotein fraction was 68 +/- 21%. This large increase in TGF-beta1 associated with the lipoprotein fraction was mainly due to association with VLDL, chylomicrons, and LDL. The lipoprotein fraction inhibits TGF-beta1 binding to the type II TGF-beta receptor extracellular domain in an ELISA and inhibits TGF-beta1 activity in the mink lung cell bioassay. We propose that sequestration of TGF beta into lipoproteins represents a novel mechanism by which TGF-beta activity in circulation may be regulated. Lipoprotein sequestration of TGF-beta may therefore contribute to the severe depression of TGF-beta activity in advanced atherosclerosis. PMID- 9392433 TI - Quantification of HDL2 and HDL3 cholesterol by the Vertical Auto Profile-II (VAP II) methodology. AB - Of the several existing methods for quantification of major subspecies of high density lipoprotein (HDL), HDL2 and HDL3, the methods based upon double precipitation are particularly useful for large-scale studies or for routine assay because of their high speed and low cost. The Vertical Auto Profile-II (VAP II) method developed in our laboratory primarily for the direct single test measurement of cholesterol (C) in all major lipoproteins, including Lp[a] and IDL, is rapid, highly sensitive, and suitable for large-scale studies. Here we describe the modification of this procedure so as to be able to quantify both HDL2- and HDL3-C in addition to all major lipoproteins without any additional assay steps, time, or cost. The VAP-II procedure was validated by comparison with four other methods using plasma samples obtained from 35 healthy subjects: 1) HDL VAP-II (a variation of the VAP-II procedure designed specifically to separate HDL subspecies); 2) dextran sulfate (DS)/Mg2+ double precipitation method performed at Northwest Lipid Research Laboratories (NWLRL), Seattle, WA; 3) 4-30% polyacrylamide-agarose (4/30 PAA) nondenaturing gradient gel electrophoresis (GGE); and 4) analytical ultracentrifugation (AUC), with both GGE and AUC performed at the Donner Laboratory, University of California at Berkeley. Both HDL2- and HDL3-C measurements by VAP-II correlated well with the measurements by all comparison methods (r for HDL3-C: HDL-VAP-II, 0.948; NWLRL, 0.947; GGE, 0.861; and AUC, 0.706, and r for HDL2-C: HDL-VAP-II, 0.867; NWLRL, 0.854; GGE, 0.885; and AUC, 0.721). The measurements of HDL2- and HDL3-C by the VAP-II method are reproducible, with the long-term between-rotor CV of 5.0% for HDL3-C and 9.0% for HDL2-C. PMID- 9392434 TI - A cDNA-dependent scintillation proximity assay for quantifying apolipoprotein A I. AB - We have developed a cDNA-dependent scintillation proximity assay (SPA) for rabbit apolipoprotein A-I that follows a classic radioimmunoassay scheme, in that antiserum and radiolabeled ligand are used in a process to quantify a source containing unlabeled ligand. To synthesize radiolabeled ligand we isolated a full length rabbit apolipoprotein A-I (apoA-I) cDNA, transcribed the corresponding RNA in vitro, and synthesized radiolabeled apoA-I by including tritiated leucine in an in vitro translation reaction. Assay conditions were established which allowed quantification of unlabeled apoA-I over a range of 0.2 to 4 nanograms with intra- and interassay coefficients of variation of 5% and 10%, respectively. Purified rabbit apoA-I, apoA-I in rabbit liver parenchymal cell conditioned media, and apoA-I contained in rabbit plasma all generated parallel titration curves. Quantification of rabbit plasma apoA-I was not affected when sheep anti-rabbit apoA-I serum was mixed with sheep anti-rabbit apoB or apoE serum; thus, the antibody need not be specific to quantify the ligand of interest. To show utility of the assay, apoA-I mass was quantified in in vitro and in vivo models displaying altered apoA-I levels. In each model apoA-I values from the cDNA dependent SPA and the established methodologies of Western blotting and electroimmunodiffusion were highly correlated. The approach outlined in this report should permit rapid development of scintillation proximity assays for other proteins given the widespread availability of full-length cDNAs. PMID- 9392435 TI - Quantitative determination of thiolipids in organic solution, in membranes, and on HPTLC plates. AB - In the presence of ortho-phthalaldehyde and glucosamine, thiolipids form fluorescent isoindole derivatives. This reaction can be used to quantify single- and double-chain mercaptans in membranes (liposomes) and micellar solutions. The lower detection limit is 100 pmol. In addition, the assay allows the detection of 1.9 nmol thiolipids on HPTLC plates and the fluorescence signal is stable for days. A minor modification of the commonly used DTNB (Ellman's) assay allows the quantification of thiolipids in organic solutions at a concentration down to 3 nmol. PMID- 9392436 TI - Mathematical theory of competitive binding assays: an exact and practical model. AB - In the usual formulation of the equations for competitive binding assays, the free concentrations of the various unlabeled ligands are approximated by the known values of their total concentrations, since the free concentrations are not easily determined. Although equations have been derived previously that give the exact solution with the free concentrations of unlabeled ligands treated as variables, these have not been useful in practice. We have devised a mathematical model for the competitive binding that which is both exact and practical for the general case of one labeled ligand, any number of unlabeled ligands, and any number of classes of binding sites. In this model, the total concentrations of unlabeled ligands are the explicit variables, instead of their free concentrations. The free concentrations of unlabeled ligands can be estimated from the model. The model is based on the law of mass action and the dilution principle, as well as a new concept, called the equivalent competitive binding principle. PMID- 9392437 TI - The unliganded mineralocorticoid receptor is associated with heat shock proteins 70 and 90 and the immunophilin FKBP-52. AB - The human mineralocorticoid receptor (MR) is a member of the steroid-thyroid hormone receptor superfamily, which includes receptors for retinoic acid, vitamin D, and other steroids, such as the glucocorticoids (which bind the glucocorticoid receptor, GR). MR and GR, the corticosteroid receptors, share significant homology and are activated by steroid binding, resulting in a conformational change, nuclear translocation, and DNA binding. Despite these similarities with GR, the MR remains less well characterized. However, protein components known to be present in the unliganded GR are also likely to be components of the heteromeric MR complex. In the current study, we investigated whether or not hsp70, hsp90, and the immunophilin FKBP-52 are present in the nonsteroid-bound MR complex, because these proteins are known to be present in the unliganded GR complex. The unliganded MR complex was assembled in vitro using reticulocyte lysate and in vivo using the baculovirus overexpression system and Spodoptera frugiperda (Sf9) cells. Western blot analysis revealed the presence of hsp70, hsp90, and FKBP-52 in the unliganded complexes, but hsp90 and FKBP-52 were not detected following exposure to aldosterone. Electrophoretic mobility shift analysis demonstrated that DNA binding of MR occurred only after treatment with aldosterone. These studies indicate that proteins associated with the unliganded GR are also present in the unliganded MR complex, and that hsp90 and FKBP-52 dissociate prior to DNA binding in a manner similar to that described for GR. Finally, the stoichiometric analysis of the proteins present within the heteromeric MR complex suggests a divergence between this receptor and the GR. PMID- 9392438 TI - Myosin phosphorylation by human cdc42-dependent S6/H4 kinase/gammaPAK from placenta and lymphoid cells. AB - The p21-activated kinase (PAK) family includes protein phosphotransferases regulated by the GTPases rho, rac, and cdc42. Sequence homology, activation mechanism, and substrate specificity suggest that the well-characterized human placenta S6/H4 kinase is a member of this family. In these studies, S6/H4 kinase purified to homogeneity from human placenta was activated in vitro by cdc42-GTP, or protease incubation and MgATP-dependent autophosphorylation. The cdc42 activated enzyme demonstrated an Mr 60,000, and shares sequence homology with the gammaPAK family. Antipeptide antibodies against one of the autophosphorylation site sequences recognized a single p60 protein in the purified placenta preparation or Jurkat cell extracts. An autophosphorylated Mr 40,000 protein, previously identified as the catalytic domain of the enzyme, was also detected by the antibody after protease activation. Crude PAK60 obtained from Mono Q chromatography of Jurkat cell extracts and purified placenta enzyme catalyzed phosphorylation of histone H4 and myelin basic protein as well as a variety of synthetic peptides previously identified as S6/H4 kinase substrates. In addition, Jurkat myosin II and the regulatory myosin light chain were phosphorylated by the Jurkat and placenta gammaPAK. Synthetic peptides were used to demonstrate that the site of light chain phosphorylation occurs at the serine which results in ATPase activation. The data suggest that human gammaPAK may regulate cell motility by a GTP-dependent and calcium-independent mechanism. PMID- 9392439 TI - Surfactant protein-A receptor-mediated inhibition of calcium signaling in alveolar type II cells. AB - Receptor-mediated inhibition of cellular activating signals is not well understood. Type II alveolar cells secrete surfactant in response to such secretagogs as terbutaline, calcium (Ca) ionophores (e.g., ionomycin [Io]), and adenosine triphosphate (ATP). A cell membrane receptor for SP-A, one of the surfactant proteins, regulates secretion by negative feedback. We used quantitative fluorescence microscopy to study the effects of SP-A on alterations in cytosolic Ca2+ ([Ca2+]i) elicited by surfactant secretagogs. Freshly isolated type II cells were loaded with Fura-2, then treated with secretagog, in the presence or absence of SP-A. Io and ATP produced biphasic increases in cytosol [Ca2+]i, reflecting first Ca2+ release from intracellular stores, and then influx through the cell membrane. Thapsigargin (TG) and Io directly initiate Ca2+ release; ATP elicits Ca2+ release via receptor-mediated mechanisms. Ca2+ release causes cell membrane Ca channels to open by as yet poorly understood mechanisms. Io itself acts as an additional Ca2+ channel. SP-A blocks much of the Ca2+ release and some of the Ca2+ influx elicited by these secretagogs. Antibody against SP-A receptor restores secretagog-induced Ca2+ fluxes from inhibition by SP-A, confirming that the inhibitory activity of SP-A is mediated through its receptor. Type II cells incubated in Ca2+-free medium plus SP-A show diminished Ca2+ release responses to TG or ATP, suggesting that the action of SP-A to prevent secretagog initiated increases in [Ca2+]i may reflect its ability to block Ca2+ release from cytoplasmic Ca stores. The feedback inhibition of surfactant secretion by SP-A may, correspondingly, be a manifestation of this effect. Because recent work suggests that TGF-beta also inhibits Ca2+ fluxes, SP A and TGF-beta could be representative of a group of physiologic regulators that act by modulating intracellular Ca signaling. PMID- 9392440 TI - G-protein-coupled receptor agonists augment adenylyl cyclase activity induced by forskolin in human corpus cavernosum smooth muscle cells. AB - The goal of this study was to investigate the synergistic effects between G protein-coupled receptor agonists and forskolin-induced activation of adenylyl cyclases, in cultured human corpus cavernosum smooth-muscle cells. Treatment of human corpus cavernosum smooth-muscle cells with forskolin (0.1-10 microM) produced an increase in cAMP synthesis in a concentration-dependent manner. Forskolin-induced adenylyl cyclase activity was markedly augmented by prostaglandin E1 (PGE1) and its metabolite, PGE0, isoproterenol, carbachol, and phenylephrine. Augmentation of forskolin-induced cAMP by PGE1, and PGE0 is probably mediated by prostaglandin E receptors (EP). Enhancement of forskolin induced cAMP synthesis by isoproterenol is mediated by beta-adrenergic receptors (beta-AR), since this activity was inhibited by propranolol. Stimulation of forskolin-induced cAMP synthesis by carbachol is attributed to activation of muscarinic acetylcholine receptors (mAChR), as demonstrated by inhibition with atropine. The augmentation of forskolin-induced cAMP synthesis by phenylephrine, an alpha1-adrenergic receptor (AR) agonist, however, was unexpected and cannot be attributed to increased intracellular Ca2+, since treatment of cells with either the Ca2+ ionophore, A23187, or 80 mM KCl did not affect forskolin-induced cAMP synthesis. Stimulation of forskolin-induced cAMP synthesis by phenylephrine is explained by its binding to beta-AR and activation of Gs protein, since this augmentation was inhibited by the beta-AR antagonist, propranolol. This observation was further supported by physiological studies in organ bath chambers, in which forskolin-induced relaxation of precontracted corpus cavernosum strips was enhanced by phenylephrine. These studies suggest that synergism between agonist-induced cAMP synthesis and forskolin is attributed to increased conformational stabilization of activated adenylyl cyclase catalytic domains by forskolin and the Gs(alpha)-subunit of activated Gs proteins. PMID- 9392441 TI - Melatonin protects against gastric ischemia-reperfusion injury in rats. AB - Lipid peroxidation and active oxygen metabolites have been suggested to play an important role in the pathogenesis of acute gastric mucosal injury induced by ischemia-reperfusion. The aim of this study was to examine the in vivo protective effects of melatonin on ischemia-reperfusion induced gastric damage in rats. The peroxidation of lipids and changes in the activities of related enzymes such as glutathione peroxidase and myeloperoxidase, as a marker of neutrophil infiltration, were also studied. Our results show that gastric injury was significantly increased after 30 min ischemia induced by clamping the celiac artery and 60 min reperfusion. Intraperitoneal administration of melatonin prevented postischemic mucosal injury. The mean ulcer indices of rats treated with 5, 10, and 20 mg kg(-1) were significantly lower (P<0.01, P<0.001) than that of control rats. These protective effects were likely in part related to a reduction of neutrophil infiltration (myeloperoxidase values). Lipid peroxidation in the stomach was increased by ischemia-reperfusion injury and this increase was inhibited by the administration of melatonin. In addition, treatment with melatonin limited the decreased glutathione peroxidase activity. The results suggest that melatonin confers a marked protection against ischemia-reperfusion gastric injury which could be due to melatonin's free radical scavenging activity and its ability to reduce neutrophil-induced toxicity. PMID- 9392442 TI - Nocturnal urinary 6-sulphatoxymelatonin excretion is decreased in primary breast cancer patients compared to age-matched controls and shows negative correlation with tumor-size. AB - In previous studies a tumor-size dependent decline of the circadian amplitude of serum melatonin was found in primary unoperated breast cancer patients, which was not due to changes of the hepatic metabolism of melatonin since its main peripheral metabolite, 6-sulphatoxymelatonin (aMT6s), showed similar serum levels. The aim of the current study was to verify these previous results by measurements of the nocturnal excretion of aMT6s in urine. The determination of aMT6s was carried out by radioimmunoassay. 17 primary unoperated breast cancer (BC) patients and 34 age-matched control patients with different types of benign gynecological diseases awaiting operation (breast diseases, n=13; ovarian diseases, n=12; and uterine diseases, n=9) were analysed. The median nocturnal urinary aMT6s excretion (22:00-6:00 hr) was significantly lower (-48%, P = 0.033) in BC patients than in controls. Controls showed a significant negative linear regression with age (r = -0.419, P = 0.014). According to multivariate linear regression analysis, BC revealed no age-dependency but a significant negative effect of increasing tumor-size on aMT6s-excretion (P = 0.036) was detected. These results confirm previous findings of a decreased pineal melatonin secretion in BC patients as well as an inverse relationship with tumor-size excluding a possible distortion due to age. The mechanisms involved are unknown but indicate that BC may lead to an impaired production of pineal melatonin. The clinical relevance of these findings from therapeutic and diagnostic point of view is discussed. PMID- 9392443 TI - A note on the time dependence of pineal gland research publications. AB - Counts of the number of publication titles containing the search truncation "pinea*" were compiled via Current Contents over the time period 1978-1994. These counts and their time dependence were examined for autocorrelations and frequency spectral components. Such analyses were carried out irrespective of either author, research laboratory, funding, or other factors. Interestingly, the results show the research publication rate in pineal studies to be linked to community history. In particular, the interactions affecting output among community members operate on a few characteristic time scales ranging from one to several years. PMID- 9392444 TI - The role of the intracellular and extracellular serotonin in the regulation of melatonin production in rat pinealocytes. AB - This study investigated whether the activation of pinealocyte beta-adrenergic receptors is involved in the regulation of serotonin (5-HT) synthesis and release, as it is for melatonin production. In addition, the role of the intra- and extra-cellular 5-HT in modulating the synthesis of melatonin induced by the beta-adrenergic agonist isoproterenol (ISO) was also studied. The incubation of dissociated pinealocytes with 0.1-10 microM ISO resulted in a concentration dependent increase of melatonin synthesis. 5-HT release and intracellular 5-HT content were increased by 0.1 and 1 microM ISO but they were reduced after ISO 10 microM. Moreover, when incubated with the tryptophan hydroxylase inhibitor p chlorophenylalanine (PCPA), the secretion of 5-HT as well as the intracellular 5 HT levels were markedly reduced in both ISO-stimulated and unstimulated conditions. Melatonin release was also inhibited by PCPA, although it responded in the expected manner to increasing concentrations of ISO. These data indicate that the release of 5-HT from pinealocytes depends on the availability of cytoplasmic 5-HT, which in turn is highly dependent on the tryptophan hydroxylase activity. In cells stimulated with moderate ISO concentrations, 5-HT release may be an important regulatory process of pineal 5-HT. After a large stimulation of N acetyltransferase (NAT) activity by ISO, the synthesis of melatonin prevails on 5 HT release, whose decrease is associated to a deficit of intracellular 5-HT. On the other hand, the present study shows that the incubation of pineal cells with high concentrations of 5-HT or with a selective 5-HT2 receptor agonist, alpha methyl-5-hydroxytryptamine, reverses partially the inhibitory effect of PCPA on the ISO-stimulated melatonin synthesis. In contrast the 5-HT2 antagonist, ketanserin, results in an inhibiton of the release of melatonin following ISO stimulation. These results suggest that released 5-HT may have a role in the full expression of the beta-adrenergically induced NAT activity and, thus, may contribute to the optimal melatonin synthesis at night. PMID- 9392445 TI - Circadian rhythm in experimental granulomatous inflammation is modulated by melatonin. AB - Biological rhythms are detected in a variety of physiological and pathological conditions in man and animals, such as rheumatoid arthritis and asthma. Here we describe a circadian rhythm in experimental infectious and non-infectious granuloma. After 30 days of BCG (Bacillus Calmette-Guerin) or nystatin inoculation in the left hind foot of C57B1/6 mice, there is an oscillation with a period of approximately 24 hr in the variation of paw thickness, indicating a circadian rhythm. The acrophase occurred during the light phase, between 9:00 and 13:00 hr, while the nadir occurred in the dark phase, between 21:00 and 01:00 hr. The vascular permeability around the granulomatous lesions was higher at 12:00 hr than at 24:00 hr. This is in agreement with the observation that the thickness of a paw with granulomatous lesion is larger during the light phase. This rhythmic variation was eliminated by either pinealectomy or superior cervical ganglionectomy, which greatly reduce melatonin levels in the blood. Nocturnal replacement of melatonin in pinealectomized mice led to the re-establishment of the circadian rhythm. Thus, the rhythm of the granulomatous lesion is due to the rhythmic melatonin release by the pineal gland. This approach opens new questions regarding the modulation of chronic inflammation in inflammatory diseases that present rhythmic symptoms throughout the day. PMID- 9392446 TI - The role of melatonin and L-tryptophan in prevention of acute gastric lesions induced by stress, ethanol, ischemia, and aspirin. AB - Melatonin, a pineal hormone, synthesized from L-tryptophan, is known to exist in the gut and to scavenge oxygen free radicals but its role in gastroprotection against acute lesions induced by various strong irritants has been little studied. In this study, we determined the effects of melatonin and L-tryptophan on gastric secretion and the formation of acute gastric lesions induced by absolute ethanol, acidified aspirin (ASA), stress, and ischemia-reperfusion (I/R). Area of gastric lesions was determined by planimetry, gastric blood flow (GBF) was measured using a H2-gas clearance technique, and blood was withdrawn for the measurement of free radicals, plasma gastrin, and melatonin concentration by specific radioimmunoassay. Intragastric (i.g.) administration of melatonin (2.5-10 mg/kg) or L-tryptophan (25-200 mg/kg) failed to affect gastric lesions by ethanol and ASA but dose-dependently reduced the lesions provoked by stress and I/R; this protective effect was accompanied by a significant rise in plasma melatonin level, GBF, and DNA synthesis and by a marked fall in blood free radicals. L-tryptophan, which significantly elevated the plasma melatonin by about 3-5-fold, also reduced the stress and I/R-induced lesions and blood levels of free radicals, while increasing the GBF, DNA synthesis, and plasma gastrin levels. Inhibition of mucosal generation of PGE2 by indomethacin abolished the protection and the rise of GBF afforded by melatonin and L-tryptophan, whereas pretreatment with N(G)-nitro-L-arginine (L-NNA), to suppress nitric oxide (NO) synthase, was without any effect. We conclude that melatonin applied exogenously in pharmacological doses and that released by the administration of its precursor, L-tryptophan, protect gastric mucosa from the damage induced by stress and I/R possibly by a mechanism involving the scavenging of free radicals and gastric hyperemia probably mediated by endogenous prostaglandin but not NO. PMID- 9392447 TI - Nocturnal urinary 6-sulfatoxymelatonin and proliferating cell nuclear antigen immunopositive tumor cells show strong positive correlations in patients with gastrointestinal and lung cancer. AB - The hormone melatonin plays a key role in coordinating neuroendocrine signals involved in the control of biological rhythms and also appears to be involved in the regulation of cellular proliferation. In this study on patients with gastrointestinal and lung cancer the nocturnal urinary excretion of 6 sulfatoxymelatonin (aMT6s) reflecting pineal melatonin production as well as immunohistochemically detectable proliferating cell nuclear antigen (PCNA) and melatonin were measured in corresponding tumor specimens (6 colorectal, 8 stomach, and 12 lung cancers). Strong positive correlations were detected between aMT6s and PCNA for the different types of tumors analysed (1 > or = Rs > or = 0.736, P < 0.01-0.0001). These findings provide support to the concept of an involvement of the pineal gland in malignancy and suggest that aMT6s-measurements may be considered as a non-invasive tool to estimate tumor cell proliferation. Negative correlations found between urinary aMT6s and melatonin in tumor cells ( 0.735 > or = Rs > or = -0.928, P < 0.01-0.0025) could be interpreted as an effort of the pineal gland to secrete melatonin to compensate for the decrease in the number of melatonin-immunopositive cells within tumor tissue where it may possess important regulatory functions. PMID- 9392448 TI - Utility of high doses of melatonin as adjunctive anticonvulsant therapy in a child with severe myoclonic epilepsy: two years' experience. AB - Recent data indicate that melatonin inhibits brain glutamate receptors and nitric oxide production, thus suggesting that it may exert a neuroprotective and antiexcitotoxic effect. Melatonin has been seen to prevent seizures in several animal models and to decrease epileptic manifestations in humans. The lack of response to conventional anticonvulsants in an epileptic child led us to use melatonin in this case. A female child who began to have convulsive seizures at the age of 1.5 months and was diagnosed as having severe myoclonic epilepsy was unsuccessfully treated with different combinations of anticonvulsants, including valproic acid, phenobarbital, clonazepam, vigabatrin, lamotrigin, and clobazam. Melatonin was thus added to the treatment. Imaging studies (CT, SPECT, and MNR), EEG recordings, blood biochemical, and hematological analyses, including measures of the circadian rhythm of melatonin, were made. The child was initially treated with various anticonvulsants. Severe neurological and psychomotor deterioration combined with increased seizure activity showed a lack of response to the treatment. At the age of 29 mon the patient was in a pre-comatose stage at which time melatonin was added to treatment. After 1 month of melatonin plus phenobarbital therapy and for a year thereafter, the child's seizures were under control. On reducing the melatonin dose after this time, however, seizures resumed and the patient's condition was re-stabilized after restoring melatonin. Prior to our attempts to reduce melatonin, all analyses, including EEG recordings and SPECT, were normal. As far as the results of neurological examination are concerned, only mild hypotony without focalization remained. Changes in the therapeutic schedules during the second year of melatonin treatment, including the withdrawal of phenobarbital, did not result in the same degree of seizure control, although progressively the child became satisfactorily controlled. At the present moment the child continues to have mild hypotony and shows attention disorder and irritability. Melatonin has proven to be useful as adjunctive therapy in the clinical control of this case of severe infantile myoclonic epilepsy. The results suggest that melatonin may have a useful role in mechanisms of neuroprotection and also indicate its use in other cases of untreatable epilepsy. Further studies using more patients and placebo-treatment would be beneficial in understanding the potential use of melatonin as a co-therapy in some cases of seizures. PMID- 9392449 TI - Protective effect of melatonin in carrageenan-induced models of local inflammation: relationship to its inhibitory effect on nitric oxide production and its peroxynitrite scavenging activity. AB - In vitro studies have demonstrated that melatonin is a scavenger of oxyradicals and peroxynitrite and an inhibitor of nitric oxide (NO) production. In the present study, we evaluated the effect of melatonin treatment in two models of acute inflammation (carrageenan-induced paw edema and pleurisy), where oxyradicals, NO, and peroxynitrite play a crucial role in the inflammatory process. Our data show that melatonin (given at 62.5 and 125 microg/paw in the paw edema model or 25 and 50 mg/kg in the pleurisy model) inhibits the inflammatory response (paw swelling, pleural exudate formation, mononuclear cell infiltration, and histological injury) in dose-dependent manner in both models. Furthermore, our data suggest that melatonin exerts an inhibitory effect on the expression of the inducible isoform of NO synthase. Melatonin also prevented the formation of nitrotyrosine, an indicator of peroxynitrite, in both models of inflammation. Taken together, the present results demonstrate that melatonin exerts potent antiinflammatory effects. Part of these antiinflammatory effects may be related to an inhibition of the expression of the inducible NO synthase, while another part may be related to oxyradical and peroxynitrite scavenging. PMID- 9392450 TI - Regional brain distribution of metallothionein, zinc and copper in toxic milk mutant and transgenic mice. AB - The regional distribution of metallothionein (MT), zinc and copper was measured in brains of transgenic MT-I overexpressor (MT-I*) mice, MT-I/MT-II gene knockout (MT-I/MT-II null) mice, and in brains of control C57BL/6J mice with normal MT expression. Toxic milk (tx) mutant mice with abnormally high MT and copper accumulation were also assessed. Although there were significant differences in MT levels (assessed by a cadmium-binding assay) in whole brain of MT-I/MT-II null and control mice (16.5 +/- 2.9 microg/g vs 25.6 +/- 7.4 microg/g), different regions of the brain (cerebral cortex, corpus striatum, hippocampus, thalamus plus hypothalamus, cerebellum, and brain stem) contained similar amounts of MT. Male MT-I* mice had significantly higher whole brain MT level than controls (35.5 +/- 8.1 microg/g vs 25.6 +/- 7.4 microg/g), and had a 2-fold higher MT level in cerebellum, but not in other brain regions. Female MT-I* mice had significantly increased MT levels in all brain regions, with the highest increase in cerebellum (3.5-fold), and the lowest increase in cortex (2-fold). MT level in whole brain of female MT-I* mice was also significantly higher than that of male MT-I* (75.2 +/- 8.0 microg/g vs 35.4 +/- 8.1 microg/g). Toxic milk mice had significantly higher MT levels in all brain regions compared to age-matched controls (51.8 +/- 10.8 microg/g vs 30.3 +/- 5.8 microg/g), while no specific region of tx mouse brain showed a preferential increase in MT. In MT-I* and MT-I/MT-II null mice, altered MT levels did not always result in altered zinc and/or copper concentrations. However, all mouse strains exhibited region-specific accumulation of zinc, with the highest level in hippocampus. In control, MT-I/MT-II null, and male MT-I* mice, the hippocampus accumulated the highest level of copper. However, MT-I/MT-II null and both male and female MT-I* mice had similar levels of copper, compared to control mice. Toxic milk mice, on the other hand, had significantly higher copper levels in cerebral cortex, corpus striatum, thalamus/hypothalamus, and brain stem, compared to control mice. Zinc levels in corpus striatum, hippocampus, and cerebellum were also significantly increased. These data indicate that, in normal control and MT-I/MT-II null mice, MT is expressed uniformly in different regions of the brain. MT-I* mice, on the other hand, exhibit regional and gender-associated change in brain MT, and tx mice have markedly increased MT, copper, and zinc levels in most brain regions. These mouse strains will be useful models in elucidating the role of MT in the pathological effects of altered zinc and copper in brain. PMID- 9392451 TI - Cosubstrates involved in the reduction of cytosolic glutathione disulfide in rat heart. AB - The functionality of glutathione (GSH), which is present in separate mitochondrial and cytosolic pools, hinges on a steady supply of reducing equivalents, provided by NADPH, to convert glutathione disulfide (GSSG) to GSH. It is believed traditionally that glucose 6-phosphate (G6-P) via the pentose phosphate pathway is the main cellular source of NADPH. The current study examined the ability of NADH- and NADPH-linked cosubstrates to support cardiac cytosolic GSSG reduction. Exogenous NADP+ was added to the incubation mixtures because of the loss of this nucleotide during homogenization. Exogenous GSSG was added to all samples to levels that were approximately 60% of total glutathione. In both the 500 x g (with mitochondria) and 10,000 x g (without mitochondria) rat heart supernatants, isocitrate supported reduction of approximately 90% of available GSSG within 10 min. Malate, pyruvate and palmitoyl carnitine did not support GSSG reduction in either supernatant. G6-P yielded GSH levels within 10 min equal to 77% of total glutathione in the 1,0000 x g supernatant and 47% in the 500 x g supernatant. The current data indicate: (1) The pentose phosphate pathway, alone, is less efficient than isocitrate at supplying reducing equivalents for cytosolic GSSG reduction; and (2) some confounding factor(s) occur in the 500 x g and reconstituted 500 x g supernatants whereby G6-P supported GSSG reduction is attenuated. PMID- 9392452 TI - The mechanisms of nickel uptake by rat primary hepatocyte cultures: role of calcium channels. AB - The present study was designed to clarify the mechanism of nickel (Ni) uptake in primary cultures of rat hepatocytes. Exposure of the hepatocytes to Ni (2-50 microM; as NiCl2) for up to 6 h was not cytotoxic, as assessed by the tetrazolium based dye (MTT) assay. Hepatocytes were treated with 10 microM NiCl2 in the absence or presence of calcium (Ca) and magnesium (Mg) (1 mM). Ni uptake was increased by 19% in medium lacking Mg or Ca and was increased by 37% in Ca- and Mg-free medium. The role of Ca channels on Ni uptake by the hepatocytes was investigated. Pretreatment with nicardipine or verapamil (200 microM), potent inhibitors of Ca channels, decreased Ni uptake by 20%. This effect was only observed when the cells were incubated in the absence of Ca. Pretreatment with vasopressin (100 nM), a well-known Ca channel agonist, significantly increased Ni uptake by the hepatocytes (24%). To determine the involvement of carrier-mediated processes on Ni uptake, the effect of temperature was also investigated. At 4 degrees C the Ni uptake was decreased by 20% compared to uptake at 37 degrees C. These results indicate that Ni uptake by the hepatocytes occurs, at least in part, through the Ca channel transport processes. Further study will be required to assess what other mechanisms are involved. PMID- 9392453 TI - Modulation of TCDD-induced fetotoxicity and oxidative stress in embryonic and placental tissues of C57BL/6J mice by vitamin E succinate and ellagic acid. AB - The ability of vitamin E succinate and ellagic acid to modulate 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD)-induced developmental toxicity and oxidative damage in embryonic/fetal and placental tissues was studied in C57BL/6J mice. Vitamin E succinate (100 mg/kg per day) and ellagic acid(6 mg/kg per day) were administered by gavage to groups of pregnant mice on days 10, 11 and 12 of gestation and 40 mg vitamin E succinate/kg or 3 mg ellagic acid/kg on day 13 of gestation. A number of animals from the vitamin E succinate and ellagic acid treated groups also received 30 microg TCDD/kg on day 12 of gestation, 2 h prior to vitamin E succinate or ellagic acid treatment. Groups of treated animals were terminated on day 14 of gestation, and the biomarkers of oxidative stress, including superoxide anion production and the induction of lipid peroxidation and DNA-single strand breaks (SSB), were determined in whole embryonic and placental tissues homogenates. Groups of treated animals were also killed on day 18 of gestation for investigation of the fetotoxic and teratogenic effects as well as effects on the placentae. Vitamin E succinate and ellagic acid significantly decreased TCDD-induced fetal growth retardation fetal death and placental weight reduction, with no significant ameliorating effects on TCDD-induced malformations including cleft palate and hydronephrosis. Vitamin E succinate treatment resulted in decreases of 77-88%, 70-87%, and 21-47% in the production of superoxide anion, lipid peroxidation and DNA-SSB, respectively, in embryonic and placental tissues, while ellagic acid caused 47-98%, 79-93%, and 37-53% decreases, respectively, in these parameters. These results indicate that TCDD-induced fetal death and fetal and placental weight reductions in C57BL/6J mice may be due to oxidative damage induced by TCDD, and ellagic acid and vitamin E succinate provide protection against those effects. Ellagic acid provided better protection than vitamin E succinate against TCDD-induced fetal growth retardation and increases in lipid peroxidation in embryonic and placental tissues. PMID- 9392454 TI - Cytochrome P450 1A1 induction in rat lymphoid tissues following in vivo and in vitro exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin requires protein kinase C. AB - The induction of cytochrome P450 1A1 (CYP1A1) is one of the most sensitive responses associated with exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and related compounds. The mechanisms that underlie this response are not completely understood, particularly in lymphoid tissues that may be used in biomarker studies in humans. CYP1A1 mRNA expression and enzyme activity (ethoxyresorufin-o-deethylase, EROD) were investigated in rat thymus and spleen and isolated thymocytes and splenocytes in culture. Thymus- or spleen-derived microsomes from rats treated in vivo with TCDD showed induced EROD activity after as little as 12 h following a single exposure to TCDD (5 microg/kg body weight). Resting rat thymocytes in culture had detectable levels of EROD activity and CYP1A1 mRNA expression which increased following in vitro exposure to > or = 0.1 nM TCDD for 24 or 48 h. Interestingly, concomitant in vitro exposure of rat thymocytes to TCDD and the mitogen concanavalin A (Con A) inhibited the induction of EROD activity, which is in contrast to the response of cultured human peripheral blood lymphocytes (Landi et al., 1994; Pharmacogenetics 4, 242 246). Resting rat splenocytes in culture had no detectable EROD activity and CYP1A1 activity could not be induced by in vitro TCDD exposure, in the presence or absence of Con A. These results suggest that the relative maturation state of the cells is important in regulating the expression of CYP1A1, since splenocytes represent a more mature population of B and T lymphocytes. TCDD-induced CYP1A1 expression in cultured rat thymocytes was inhibited by the addition of calphostin C, a specific protein kinase C (PKC) inhibitor, suggesting a role for PKC as a second messenger in the CYP1A1 induction pathway. In vivo co-exposure with phorbol-myristate-acetate (PMA) and TCDD also inhibited CYP1A1 induction. Again, suggesting a role for PKC in CYP1A1 induction. Together, these results indicate that relative lymphocyte maturation state and the PKC pathway are important factors in regulating the expression of CYP1A1. PMID- 9392455 TI - Dichloroacetic acid pretreatment of male and female rats increases chloroform metabolism in vitro. AB - The role of metabolism in dichloroacetic acid (DCA) potentiation of CHCl3 hepatotoxicity was investigated. Male and female Sprague-Dawley rats were given three doses (09:00, 16:00 and 09:00 the next morning) of DCA (each 2.45 mmol/kg) by gavage. The rats were euthanized 3 h after the last dose, hepatic microsomes were prepared and 14CHCl3 metabolism was measured in vitro. The binding of 14CHCl3-derivatives to microsomal proteins and lipids was increased 65 and 100%, respectively, in DCA-treated rats compared to their respective NaCl-treated controls. The formation of CO2 (nmol 14CO2/incubation) was significantly elevated in DCA-treated rats compared to controls (10.4 vs 6.3 in males; 10.8 vs 6.1 in females). DCA treatment decreased the apparent Michaelis constant (Km app) for conversion of 14CHCl3 to 14CO2 in rats (0.665 vs 0.415 mM in males, P < 0.05; 0.161 vs 0.081 in females). 14CO2 production and 14C binding were observed under N2 atmosphere, indicating that reactive metabolites of 14CHCl3 were formed by oxidation as well as reduction. Male and female rats metabolized CHCl3 differently. The Km app for CO2 production was up to 5-fold higher in the males than in the females, regardless of DCA treatment. Inhibition by SKF 525-A and piperonyl butoxide was gender dependent in both control and DCA-treated groups. The results showed, that increased bioactivation of CHCl3 by DCA treatment is one element in the DCA-CHCl3 interaction. PMID- 9392456 TI - Dichloroacetic acid pretreatment of male and female rats increases chloroform induced hepatotoxicity. AB - Dichloroacetic acid (DCA) and chloroform (CHCl3) are both major by-products of drinking water chlorination and DCA increases the hepatotoxicity of CHCl3. In this study, we further characterized this effect and investigated DCA-induced alterations of CHCl3 disposition and metabolism as a possible mechanism for this interaction. Both adult male and female Sprague-Dawley rats were gavaged with three doses (09:00, 16:00 and 09:00 the next morning) of DCA (each 2.45 mmol/kg), then challenged with an i.p. injection of CHCl3 (3.12 or 9.35 mmol/kg). Hepatic damage was assessed 24 h after CHCl3 administration as increased alanine aminotransferase (ALT), ornithine carbomyl transferase (OCT) and bilirubin in plasma. In a separate experiment, rats were pretreated with DCA or were given 14CHCl3 at the same dosages. The disposition of 14C in various tissues and covalent binding of 14CHCl3-derivatives to liver proteins and lipids were determined 1 h later. CHCl3-induced hepatotoxicity was significantly more severe in DCA-pretreated groups. ALT and OCT were more markedly elevated in DCA + CHCl3 (3.12 mmol/kg) groups than NaCl +CHCl3 animals. Plasma bilirubin content was elevated only in DCA + CHCl3 groups and females were more susceptible to this effect. The responses of rats to DCA treatment were somewhat gender-different. DCA treatment increased total cytochrome P450 in females, but not in males. Hepatic glutathione concentration was elevated in males after DCA treatment, but not in females. In the present study we confirmed that DCA pretreatment potentiates the CHCl3-hepatotoxicity of both male and female rats. However, there was little evidence that DCA pretreatment significantly affected CHCl3 disposition or increased CHCl3 binding in vivo. PMID- 9392457 TI - Dietary alpha-tocopherol supplementation on antioxidant defenses after in vivo iron overload in rats. AB - The effect of dietary alpha-tocopherol (alpha-T) supplementation on iron overload dependent oxidative damage was studied. Male Wistar rats were fed diets supplemented with 2.5% carbonyl iron and/or 200 mg/kg of alpha-tocopheryl acetate for 6 weeks. Oxidation of lipids and proteins were increased by iron overload in rat liver, and alpha-T dietary supplementation effectively prevented these effects. Iron overload decreased both, catalase and Mn-superoxide dismutase activities by 49 and 54%, respectively, with no effect on glutathione peroxidase activity. Alpha-T supplementation did not prevent the inhibition measured in catalase and Mn-superoxide dismutase activities. Iron dietary excess had no effect on liver alpha-T and ubiquinol 9 (UQ9) content. Ubiquinol 10 (UQ10) content after iron overload was decreased by 58 and 54% in whole liver and liver mitochondria, respectively. Alpha-T supplementation led to significant increases in alpha-T, UQ9 and UQ10 content in liver, as compared to control values, and partially prevented the decrease in UQ10 content due to iron excess. The results presented here indicate that initial stages of iron overload led to oxidative damage in liver (evaluated in terms of lipid and protein oxidation) with a decline in antioxidant defenses. alpha-T supplementation affected the liver content of lipid soluble antioxidants, suggesting a concerted antioxidant response at the cellular level to modulate the effect of excess iron availability. PMID- 9392458 TI - The current role of the barium examination of the small intestine. PMID- 9392459 TI - The role of CT and MR in imaging the complications of sickle cell disease. AB - Sickle cell disease is the most common inherited haemoglobinopathy described. Complications of sickle cell disease (SCD) are due to chronic haemolysis of fragile red cells or secondary to vascular occlusion by sickled red cells with subsequent tissue infarction. Traditionally plain film radiography has been the mainstay in the assessment of patients with SCD, but increasingly magnetic resonance (MR) imaging and computed tomography (CT) are being used. In this review the imaging features of a range of complications of SCD are demonstrated with particular emphasis on CT and MR. PMID- 9392460 TI - Three-dimensional computed tomography bronchoscopy using clinical datasets: a comparison with fibreoptic bronchoscopy. AB - OBJECTIVE: To assess three-dimensional computed tomography 'bronchoscopic' (3 DCTB) reconstruction of routine CT data as a non-invasive method of airway visualization, and compare it with fibreoptic bronchoscopy (FOB). METHODS: Fourteen datasets were acquired from 13 patients undergoing both FOB and CT examination of the chest. Standard continuous volume CT using 6 mm collimation and clinical FOB techniques were employed. Images were obtained from 3-DCTB reconstructions by segmentation and surface recognition algorithms generating surface rendered 'bronchoscopic views'. These were scored for technical quality and anatomical detail. The most distal bronchi seen in left upper and right lower lobes were recorded for FOB and 3-DCTB. RESULTS: On FOB, the subsegmental bronchi were seen in right lower and in left upper lobe in 10/14 cases and 4/14 cases, respectively. Visualization of the subsegmental airways was not achieved with 3 DCTB, as they could not be identified with confidence. 3-DCTB never achieved a more distal view than obtained by FOB. Using 3-DCT, the right, lower lobe segmental bronchi were seen in 10/14 cases, and lobar bronchus in 14/14 cases (two occluded). In the left upper lobe, 3-DCT showed segmental bronchi in 6/14 cases, lobar bronchus in 11/14 cases (one occluded) and the left main bronchus appeared occluded in 3/14 cases. Overall, technical quality and anatomical detail scores of the carina and proximal bronchi ranked significantly higher than views of segmental bronchi. CONCLUSIONS: 3-DCTB cannot routinely replace FOB for inspection of major and segmental bronchi. Subsegmental bronchi cannot be adequately demonstrated by 3-DCTB using 6 mm collimation datasets. PMID- 9392462 TI - Prognostic indicators in acute pancreatitis: CT vs APACHE II. AB - PURPOSE: To investigate the correlation between established contrast-enhanced computed tomography (CECT) criteria of disease severity in acute pancreatitis and the APACHE (Acute Physiology and Chronic Health Evaluation) II score and to assess the utility of each as prognostic indicators in acute pancreatitis. MATERIALS AND METHODS: Over a 1-year period, prospective, consensus interpretation of the CECTs of 35 consecutive inpatients was performed with determination of the CECT grade, degree of necrosis, and severity index. The APACHE II score was calculated within 24 h of CECT. Multiple clinical endpoints were recorded: local complications (pseudocyst, abscess, or acute fluid collections requiring urgent surgical or radiological intervention), systemic disease (intensive care unit admission), and duration of hospitalization. Statistical analysis was performed to determine correlations. RESULTS: No statistically significant correlation existed between the APACHE II score and CECT grade, the degree of necrosis, or the CECT severity index. Only the CECT grade and severity index correlated significantly with the occurrence of local complications (P = 0.0035 and 0.0048, respectively). The APACHE II score was superior to the CECT grade as a predictor of the need for ICU admission (P = 0.022 vs P = 0.035), and no other CECT criteria was a significant predictor of ICU admission. CONCLUSION: The preferred clinical and imaging prognostic measures in acute pancreatitis, the APACHE II score and CECT criteria, do not correlate with one another in the commonly encountered, mixed primary and tertiary care population. The mathematical integration of CECT criteria and the APACHE II score fails to yield a predictor of outcome superior to the use of any one measure alone. CECT criteria best define local anatomic abnormality, and are superior to the APACHE II score as predictors of local complications. The APACHE II score is superior to all CECT criteria as an indicator of systemic disease severity (reflected in the need for ICU admission). The most effective initial triage would be immediate APACHE II calculation. Further use of imaging vs clinical assessment can then be individualized. PMID- 9392461 TI - Radiological features of pulmonary tuberculosis in 963 HIV-infected adults at three Central African Hospitals. AB - Tuberculosis is one of the most important infectious complications in human immunodeficiency virus (HIV)-infected individuals in sub-Saharan Africa. In this radiological study, we detail the chest radiographic findings of Zairean and Zambian adults with a diagnosis of AIDS and tuberculosis as seen at three Central African Hospitals. Between 1992 and 1995, consecutive chest radiographs of 963 HIV-infected adults aged between 16 years and 56 years with microbiologically confirmed tuberculosis (TB) were reviewed: (1) 362 adults from Sendwe General Hospital, Lubumbashi, Zaire, (2) 175 from Mama Yemo Hospital, Kinshasa, Zaire, and (3) 426 adults from The University Teaching Hospital (UTH), Lusaka, Zambia. During the same period consecutive chest radiographs from 1000 age-matched HIV negative adults with tuberculosis were collected for comparative purposes. Comparison of the two groups showed that the HIV-infected group of patients with tuberculosis had a significantly higher proportion of lymphadenopathy (26% vs 13%; P = 0.001), pleural effusions (16% vs 6.8%; P = 0.001), miliary shadowing (9.8% vs 5%; P = 0.001), an interstitial pattern (12% vs 7%; P = 0.01) and consolidation (10% vs 3%; P = 0.001). There was significantly less cavitation (33% vs 78%; P = 0.001) and atelectasis (12% vs 24%; P = 0.001) seen in the HIV positive group compared to the HIV-negative group of patients. These patterns of radiographic changes were consistently seen across all three hospital sites. The radiographic appearances in HIV-infected individuals with TB is discussed. PMID- 9392463 TI - End stage renal transplant failure: allograft appearances on CT. AB - INTRODUCTION: Failed renal allografts often are left in situ in patients who revert to chronic dialysis therapy or who undergo retransplantation. These patients may be investigated with computed tomography (CT) imaging for allograft related or other abdominopelvic disease. This study describes the appearances of failed renal transplants on CT. METHODS: A retrospective study was made of the clinical records and CT findings on 25 studies in 14 patients, 5-156 months (average, 44 months) following allograft failure. CT studies were reviewed for allograft position, size, shape, attenuation value, calcification, cyst formation, related abdominopelvic findings and the presence of other allografts. Correlation was made with clinical findings in all patients and with pathological findings in six. RESULTS: Global shrinkage was noted in eight failed allografts, all of which were asymptomatic. Enlargement of two failed allografts was due to symptomatic acute infarction of the allograft in one patient and subacute haemorrhagic infarction simulating a tumour mass in another. CT attenuation values in individual allografts varied markedly due to fatty replacement, hydronephrosis, haemorrhage or dense calcification. Both a failed longstanding and a functioning more recently placed renal allograft were present in seven patients, four of whom had acute complications related to the more recently transplanted kidney. Two of six calcified allografts were mistaken for opacified bowel on CT. CONCLUSION: A wide spectrum in size, shape and attenuation values may be detected in failed renal allografts by CT. These organs may be the site of acute disease despite their lack of physiological function or may be diagnostically confusing findings in patients with acute disease related to more recently transplanted organs. PMID- 9392464 TI - Breast MR and the appearance of the normal and abnormal nipple. AB - AIM: To identify the magnetic resonance (MR) morphological and enhancement characteristics of the normal and diseased nipple. MATERIALS AND METHODS: The MR appearances of the nipple in 35 patients with known primary breast cancer who went on to mastectomy was reviewed by two radiologists (blinded to the clinical, mammographic and histopathological information) and correlated with histology. The appearance of the contra-lateral nipple in 31 patients was reviewed and compared with that of the affected side. MR was performed at 1.0T using a receive only double breast coil in 33 patients and a single breast coil in two patients. Three dimensional (3-D) T1-weighted gradient-echo images were made before and immediately after a fast hand injection of gadolinium-DTPA (0.1 mmol/kg). RESULTS: Twenty-six breasts had histopathologically normal nipples and retroareolar tissue, four had evidence of tumour within the nipple and four had retroareolar tumour but with nipple sparing. Fifteen normal nipples were everted and 11 were inverted (flat). All showed superficial linear dermal enhancement above a non-enhancing zone in the nipple areolar complex. Linear or patchy enhancement deep to the non-enhancing zone was seen in four breasts and linear enhancement through the non-enhancing zone was seen in two. The nipple in all four breasts with tumour involvement showed increased thickening/bulkiness and enhancement of the nipple-areolar complex and retroareolar tissue. The four breasts with retroareolar tumour and nipple sparing showed increased thickening and enhancement of the retroareolar tissue only. There was one false positive result on breast MR for retroareolar tumour involvement. In this case the abnormally enhancing retroareolar tissue adjacent to the focal mass was shown to be an area of sclerosing adenosis on histology. The 31 contra-lateral nipples had the characteristic 'normal' appearance and when compared with its normal ipsilateral nipple showed marked symmetry. When compared with its abnormal ipsilateral nipple showed marked asymmetry. CONCLUSION: MR of the breast can show nipple involvement even when clinically unsuspected. This is important for treatment planning of breast cancer, in particular nipple preserving surgery. PMID- 9392465 TI - Reduction of adverse events in MRI of the breast by personal patient care. AB - PURPOSE: To determine the difference in anxiety reactions in patients undergoing standard (non-breast) magnetic resonance imaging (MRI) compared to breast magnetic resonance imaging (MRM) and to evaluate the influence of patient information before the breast imaging examination on the rate of premature termination of the procedure. MATERIALS AND METHODS: Over 2 years, 5837 non breast and 336 breast magnetic resonance examinations were performed at our institution. One group of breast MRM patients (n = 144) received detailed information and a second group (n = 189) received only routine information before MRI. The rates of premature termination were recorded for all groups. RESULTS: In 0.5% (27/5837) of patients undergoing standard MRI examinations the study had to be stopped prematurely. Of the breast MRM patients, those who had received only routine information had a significantly higher rate of premature termination when compared to the better-informed patients and those undergoing standard MRI (5.5%, 10/189, P= 0.01). A significantly lower rate of premature termination occurred in the better-informed breast group (0%, 0/144). CONCLUSION: MRM is associated with an increase in patient anxiety and higher rates of incomplete examination than other MR procedures. We recommend careful patient preparation including detailed verbal information before MRM and support during the procedure to obtain optimal patient compliance. PMID- 9392466 TI - Radiological features of papillary carcinoma of the breast. AB - Seventeen patients with papillary carcinoma of the breast were analysed with respect to the radiological findings by three experienced breast radiologists. The most frequent mammographic appearance of papillary tumours was of an ill defined (70%) and lobulated (60%) mass and at ultrasound as a well-defined (76%), inhomogeneous (62%) and hypoechoic (92%) lesion. Histopathological subtypes of encysted papillary carcinoma, encysted papillary carcinoma with an invasive focus and invasive papillary carcinomas could not be predicted from the radiological features, although invasive tumours tended to be larger at presentation than the other subtypes. PMID- 9392467 TI - Technical report: double-phase technetium-99m-sestamibi parathyroid imaging with an in-field photopenic marker for better mapping. AB - The author introduces a photopenic marker that was used to delineate better anatomic mapping in a double-phase technetium-99m-sestamibi parathyroid imaging. This home-marker was placed on a patient within the imaging field to increase reproducibility of the images: early versus delayed images. PMID- 9392469 TI - Case report: an unusual interval breast cancer masquerading as a simple cyst. PMID- 9392468 TI - Case report: Sylvian fissure meningioma without dural attachment in a 4-year-old child. PMID- 9392470 TI - Case report: imaging features of pleural lymphoma complicating a long-standing pyothorax. PMID- 9392471 TI - Case report: portal hypertension causing massive enlargement of an accessory spleen -- a rare cause of splenic pseudotumour. PMID- 9392472 TI - Administration of intravascular contrast media to children: current practice in the UK. PMID- 9392473 TI - Fluoroscopically guided retrograde ureteric stent retrieval and replacement. PMID- 9392474 TI - What is acceptable radiological performance? PMID- 9392475 TI - Hypatia: change, limits, and interconnectedness. PMID- 9392476 TI - Augmentation of insulin release by glucose in the absence of extracellular Ca2+: new insights into stimulus-secretion coupling. AB - Glucose stimulates insulin secretion in the pancreatic beta-cell by means of a synergistic interaction between at least two signaling pathways. One, the K(ATP) channel-dependent pathway, increases the entry of Ca2+ through voltage-gated channels by closure of the K(ATP) channels and depolarization of the beta-cell membrane. The resulting increase in [Ca2+]i stimulates insulin exocytosis. The other, a K(ATP) channel-independent pathway, requires that [Ca2+]i be elevated and augments the Ca2+-stimulated release. These mechanisms are in accord with the belief that glucose-stimulated insulin secretion has an essential requirement for extracellular Ca2+ and increased [Ca2+]i. However, when protein kinases A and C are activated simultaneously, a large effect of glucose to augment insulin release can be seen in the absence of extracellular Ca2+, under conditions in which [Ca2+]i is not increased, and even when [Ca2+]i is decreased to low levels by intracellular chelation with BAPTA. In the presence or absence of Ca2+, there are similarities in the characteristics of augmentation of insulin release that suggest that only one augmentation mechanism may be involved. These similarities include time course, glucose dose-responses, augmentation by nutrients other than glucose such as alpha-ketoisocaproate (alpha-KIC), and augmentation by the fatty acids palmitate and myristate. However, augmentation in the presence and absence of Ca2+ is distinctly different in GTP dependency. Therefore, exocytosis under these two conditions appears to be triggered differently-one by Ca2+ and the other by GTP or a GTP-dependent mechanism. The augmentation pathways are likely responsible for time-dependent potentiation of secretion and for the second phase of glucose-stimulated insulin release. PMID- 9392477 TI - Tumor necrosis factor-alpha induces apoptosis of human adipose cells. AB - Tumor necrosis factor-alpha (TNF-alpha) production by adipocytes is elevated in obesity, as shown by increased adipose tissue TNF-alpha mRNA and protein levels and by increased circulating concentrations of the cytokine. Furthermore, TNF alpha has distinct effects on adipose tissue including induction of insulin resistance, induction of leptin production, stimulation of lipolysis, suppression of lipogenesis, induction of adipocyte dedifferentiation, and impairment of preadipocyte differentiation in vitro. Taken together, these effects all tend to decrease adipocyte volume and number and suggest a role for TNF-alpha in limiting increase in fat mass. The aim of the present study was to determine if TNF-alpha could induce apoptosis in human adipose cells, hence delineating another mechanism by which the cytokine could act to limit the development of, or extent of, obesity. Cultured human preadipocytes and mature adipocytes in explant cultures were exposed in vitro to human TNF-alpha at varying concentrations for up to 24 h. Apoptosis was assessed using morphological (histology, nuclear morphology following acridine orange staining, electron microscopy) and biochemical (demonstration of internucleosomal DNA cleavage by gel electrophoresis and of annexin V staining using immunocytochemistry) criteria. In control cultures, apoptotic indexes were between 0 and 2.3% in all experiments. In the experimental systems, TNF-alpha induced apoptosis in both preadipocytes and adipocytes, with indexes between 5 and 25%. Therefore, TNF-alpha induces apoptosis of human preadipocytes and adipocytes in vitro. In view of the major metabolic role of TNF-alpha in human adipose tissue, and the knowledge that adipose tissue is dynamic (with cell acquisition via preadipocyte replication/differentiation and cell loss via apoptosis), these findings describe a further mechanism whereby adipose tissue mass may be modified by TNF-alpha. PMID- 9392478 TI - Regulation of hepatic glutaminase in the streptozotocin-induced diabetic rat. AB - The liver of diabetic animals removes increased quantities of glutamine. We therefore examined factors that affect hepatic glutaminase activity in hepatocytes and mitochondria. Glutamine use, through glutaminase, was measured in isolated rat hepatocytes by monitoring the production of 14CO2 from [1 (14)C]glutamine. Hepatocytes from streptozotocin-induced diabetic rats use glutamine more rapidly than do hepatocytes from normal or insulin-maintained diabetic rats. Glutamine use in all of these hepatocytes was stimulated by glucagon and epinephrine. Glutaminase activity, assayed in broken mitochondrial membranes, was increased approximately 2.5-fold in diabetic rats. The sensitivity of glutaminase, measured in intact liver mitochondria, to phosphate was markedly left-shifted in mitochondria from diabetic rats compared with those from controls. In fact, glutaminase was increased 10-fold at 2.5 mmol/l phosphate compared with controls. This increased sensitivity of glutaminase to physiological concentrations of phosphate is characteristic of its hormonal activation. Therefore, activation of glutaminase plays a major role in diabetes and is as important as increases in its total enzyme amount in determining the increased glutamine uptake in diabetes. PMID- 9392479 TI - Effect of captopril, losartan, and bradykinin on early steps of insulin action. AB - Insulin initiates its metabolic and growth-promoting effects by binding to the alpha subunit of its receptor, thereby activating the kinase in the beta subunit. This event leads to tyrosyl phosphorylation of its cytosolic substrate, insulin receptor substrate 1 (IRS-1), which in turn associates with and activates phosphatidylinositol (PI) 3-kinase. The clinical use of ACE inhibitors has been associated with increased insulin sensitivity. However, the exact molecular mechanism is unknown. In the present study, we examined the phosphorylation status of the insulin receptor and IRS-1, as well as the association between IRS 1 and PI 3-kinase in the liver and muscle of 20-month-old rats treated acutely with captopril, using immunoprecipitation with antipeptide antibodies to the insulin receptor and IRS-1, and immunoblotting with antiphosphotyrosine and anti PI 3-kinase antibodies. Insulin stimulation increased receptor autophosphorylation to 462 +/- 253% (P < 0.05) in the liver and 697 +/- 78% (P < 0.001) in the muscle of ACE inhibitor-treated rats. There were also increases to 250 +/- 17% (P < 0.001) and 280 +/- 50% (P < 0.05) in the insulin-stimulated IRS 1 phosphorylation levels in the liver and muscle, respectively, of animals treated with captopril. The insulin-stimulated IRS-1 association with PI 3-kinase rose to 305 +/- 20% (P < 0.001) in liver and 267 +/- 48% (P < 0.05) in muscle. Losartan, an ANG receptor blocker, had no significant effect on insulin stimulated IRS-1 phosphorylation in both tissues. The acute administration of bradykinin increased insulin-stimulated tyrosine phosphorylation of the insulin receptor and IRS-1 in the liver and muscle. These data demonstrate that ACE inhibitors modulate the early steps of insulin signaling, and that this effect may be simulated by the administration of bradykinin. PMID- 9392480 TI - Sensitization to insulin induced by beta,beta'-methyl-substituted hexadecanedioic acid (MEDICA 16) in obese Zucker rats in vivo. AB - Beta,beta'-methyl-substituted hexadecanedioic acid (MEDICA 16) consists of a nonmetabolizable long-chain fatty acid designed to probe the effect exerted by fatty acids on insulin sensitivity. The effect of MEDICA 16 was evaluated in insulin-resistant Zucker (fa/fa) rats in terms of liver, muscle, and adipose tissue response to clamped euglycemic hyperinsulinemia in vivo. Nontreated Zucker rats were insulin resistant, maintaining basal rates of total-body glucose disposal, glucose production in liver, free fatty acid (FFA) flux into plasma, and FFA reesterification in adipose tissue, irrespective of the insulin levels induced. MEDICA 16 treatment resulted in an insulin-induced decrease in hepatic glucose production, together with an insulin-induced increase in total-body glucose disposal. Intracellular reesterification of lipolysed FFA in adipose tissue was specifically activated by MEDICA 16, resulting in a pronounced decrease in FFA release, with a concomitant decrease in plasma FFA. In conclusion, MEDICA 16 treatment results in the sensitization of liver, muscle, and adipose tissue to insulin in an animal model for obesity-induced insulin resistance. PMID- 9392481 TI - Effect of insulin on GLUT4 cell surface content and turnover rate in human skeletal muscle as measured by the exofacial bis-mannose photolabeling technique. AB - Insulin-stimulated glucose transport across the skeletal muscle cell membrane is a major regulatory step in postprandial glucose disposal. To estimate the total molar concentration of GLUT4 as well as the turnover rate of GLUT4 in human vastus lateralis muscles at the cell surface in the basal state and after insulin exposure, we have applied the sensitive exofacial bis-mannose photolabeling technique on in vitro incubated human skeletal muscle strips from healthy subjects. In addition, we have measured 3-O-methylglucose transport in other muscle strips prepared from the same surgically removed human skeletal muscle biopsies to compare glucose transport with cell surface level of GLUT4. Maximal in vitro insulin stimulation (2,400 pmol/l) resulted in a twofold increase compared with basal in both surface GLUT4 content (0.38 +/- 0.05 vs. 0.19 +/- 0.03 pmol/g wet muscle wt, P < 0.005) and 3-O-methylglucose transport (1.24 +/- 0.13 vs. 0.63 +/- 0.08 pmol x ml(-1) x h(-1), P < 0.005). The insulin-induced increment in 3-O-methylglucose transport was strongly correlated with the insulin induced increase in cell surface GLUT4 content (r2 = 0.91; P < 0.005). The calculated turnover rate of human skeletal muscle GLUT4 amounted to approximately 8 x 10(4) min(-1) at 35 degrees C and was unaffected by insulin. In conclusion, maximal in vitro insulin stimulation of vastus lateralis muscle strips from healthy subjects resulted in a twofold rise in glucose transport as well as in cell surface content, whereas the turnover rate of GLUT4 was unaffected by insulin under the chosen experimental conditions. PMID- 9392482 TI - Reduction in diabetes incidence in an I-Ag7 transgenic nonobese diabetic mouse line. AB - Susceptibility to IDDM is strongly associated with major histocompatibility complex (MHC) class II genotypes. Nonobese diabetic (NOD) mice develop a similar autoimmune diabetes and have a unique MHC class II I-A allele that is required for the development of diabetes. A number of groups have shown that the introduction of resistant MHC class II alleles as transgenes into the NOD mouse protects from diabetes. We made control transgenic NOD mice, expressing their own I-Abetag7 molecule as a transgene. One of two lines of these mice showed a reduced incidence of diabetes, without any change in T-cell proliferative response to a number of diabetes autoantigens or any change in insulitis severity. This line developed a subtle decrease in the percentage of splenic B cells that progressed with age. This defect was not associated with any other phenotypic abnormalities. Our findings suggest that assessment of splenic B-cell number is necessary in interpretation of the effects of MHC class II transgenes on the development of diabetes in the NOD mouse. PMID- 9392483 TI - Retardation or acceleration of diabetes in NOD/Lt mice mediated by intrathymic administration of candidate beta-cell antigens. AB - A single injection of syngeneic islet cells into the thymus of 4-week-old NOD/Lt female mice strongly retards diabetogenesis. The present study used the intrathymic route of antigen administration to compare the relative efficacy of peptides/proteins derived from two major candidate pancreatic beta-cell autoantigens, insulin and GAD65, to modulate diabetogenesis. Intrathymic administration of insulin B chain or recombinant human GAD65 significantly suppressed diabetogenesis during a 20-week follow-up period, whereas no protection was mediated by either insulin A chain or a synthetic peptide (A2) derived from it. Quite unexpectedly, two GAD65-derived peptides near the COOH terminus (p34 and p35) accelerated diabetes onset. Semiquantitative reverse transcription-polymerase chain reaction analysis was performed on cDNAs from isolated islets or whole pancreases of NOD/Lt females 4 weeks after intrathymic injections. Protection mediated by intrathymic administration with either intact islet cells or GAD65 were correlated with an upregulation of mRNA for T-helper 2 (Th2)-associated cytokines (interleukin [IL]-4, IL-10), concomitant with downregulation of Th1-associated interferon (IFN) transcripts (all normalized to T-cell receptor Cbeta transcripts) in islet-infiltrating lymphocytes. Protection mediated by the intrathymic administration of insulin B chain, however, correlated only with a modest upregulation of IL-4 and IL-10 transcript levels, and no diminution in IFN-gamma transcripts. In contrast, the diabetes accelerating GAD65 p34 and p35 peptides were not associated with an immune deviation, expressing levels of IFN-gamma characteristic of islet-infiltrating lymphocytes in vehicle-injected NOD controls. Hence, Th1-to-Th2 immune deviation provides only a partial explanation for peptide immunotherapy of diabetes in NOD mice. The finding that certain peptides can accelerate rather than retard diabetogenesis as a function of route and age of administration adds a cautionary note to this type of therapy. PMID- 9392484 TI - Long-term function (6 years) of islet allografts in type 1 diabetes. AB - Eight type 1 diabetic patients, ages 29-41 years, with mean diabetes duration of 23 years (range 18-29 years) received islet transplants from 1 to 5 donors. Seven patients had stable kidney allografts 1-11 years before the islet transplant, and one patient had a simultaneous islet-kidney allograft. Patients' blood glucose control was poor as reflected by the mean +/- SD HbA1c of 9.1 +/- 1.7% before transplant. Of the first three patients, two (1 and 3) achieved insulin independence for 36 and 38 days, respectively. Two recipients rejected their islet grafts within 1 month (2 and 8) and therefore were excluded from analysis. The HbA1c and insulin requirement of the six remaining patients who had persistent islet function for more than 60 days was significantly reduced from 9.3 +/- 1.9 to 6.4 +/- 1.0% (P = 0.002) and from 0.75 +/- 0.15 to 0.35 +/- 0.12 U x kg(-1) x day(-1) (P < 0.001), respectively. The two patients with the longest graft survival (4 and 6) achieved a normalization or near-normalization of their HbA1c levels during 6 years in the absence of severe episodes of hypoglycemia. As demonstrated by a decline in C-peptide response during Sustacal challenge tests over a 6-year period, there was a diminution of islet allograft function over time, despite persistence of normal or near normal HbA1c. We concluded that transplantation of allogeneic islets with an islet mass comparable with whole or segmental pancreas transplants in type 1 diabetic patients can result in long term islet allograft function; further, we concluded that, in conjunction with small dosages of exogenous insulin, a functioning islet allograft can result in near-normalization of blood glucose levels and significant improvement in HbA1c. The occurrence of severe hypoglycemic episodes observed for patients in the Diabetes Control and Complications Trial was not observed in recipients with functioning islet transplants, despite the continuous need for exogenous insulin therapy to sustain normal HbA1c over the 6-year follow-up. The significant improvement in metabolic control observed for the patients described in this study, and the potential to significantly decrease or halt the progression of diabetic complications, support the continued application of islet allotransplantation as a treatment modality for type 1 diabetic patients. PMID- 9392485 TI - The relation of proinsulin, insulin, and proinsulin-to-insulin ratio to insulin sensitivity and acute insulin response in normoglycemic subjects. AB - Plasma levels of proinsulin and its conversion intermediates are elevated in NIDDM patients. Recent studies have suggested that proinsulin levels are also increased relative to insulin levels in subjects who subsequently develop NIDDM. This may be due to insulin resistance or a defect in proinsulin processing or insulin secretion. If insulin resistance is the trigger, the proinsulin-to insulin ratio would be higher in insulin-resistant subjects than in insulin sensitive subjects. We examined the association of fasting proinsulin, 32,33 split proinsulin, and the proinsulin-to-insulin ratio with insulin sensitivity (SI), estimated by a frequently sampled intravenous glucose tolerance test and the minimal model in 138 normoglycemic subjects ages 53-61 years. We also investigated the relation of proinsulins and the proinsulin-to-insulin ratio to acute insulin response (AIR). Fasting specific insulin (r = -0.64), intact proinsulin (r = -0.43), and 32,33 split proinsulin (r = -0.54) concentrations were inversely correlated and the proinsulin-to-insulin ratio positively (r = 0.31) correlated with SI (P < 0.001). Fasting specific insulin (r = 0.64), intact proinsulin (r = 0.35), and 32,33 split proinsulin (r = 0.45) concentrations were positively correlated and proinsulin-to-insulin ratio (r = -0.40) inversely correlated with AIR (P < 0.001). The proinsulin-to-insulin ratio increased by increasing levels of SI (quartiles of SI from low to high: 0.048, 0.078, 0.078, 0.068; P = 0.012) and decreased by increasing AIR (quartiles of AIR from low to high: 0.088, 0.068, 0.058, 0.058; P = 0.005). These associations were independent of age, sex, BMI, and waist-to-hip ratio. Furthermore, the relation between the proinsulin-to-insulin ratio and AIR was independent of SI. In conclusion, in normoglycemic subjects, insulin resistance (low SI) was associated with a low rather than a high proinsulin-to-insulin ratio. Subjects who maintained normoglycemia with a low AIR had an increased proinsulin-to-insulin ratio compared with those who needed high AIR to maintain normoglycemia. These results suggest that, in subjects with normal glucose tolerance, insulin resistance does not induce increased proinsulin relative to insulin secretion, but rather is associated with enhanced processing of proinsulin. PMID- 9392486 TI - Oscillations in activities of enzymes in pancreatic islet subcellular fractions induced by physiological concentrations of effectors. AB - Glucose, the most potent insulin secretagogue, stimulates insulin secretion by aerobic glycolysis, but other secretagogues stimulate insulin release exclusively by mitochondrial metabolism. It is well known that in the intact pancreatic beta cell, either kind of secretagogue can induce oscillations in metabolism (e.g., glycolysis, ATP/ADP, NAD(P)/NAD(P)H ratios) that occur with a periodicity similar to oscillations in membrane electrical potential and insulin secretion. In this study, pancreatic islet cytosol or mitochondrial fractions were incubated in the presence of physiological concentrations of substrates. Repeated additions of physiological effectors caused oscillations in the activities of the three enzymes studied. Succinate dehydrogenase activity in islet mitochondrial extracts was made to oscillate by adding oxaloacetate (5 micromol/l) to inhibit the enzyme. The enzyme was reactivated by adding acetyl-CoA (3 micromol/l), which combines with oxaloacetate in the citrate synthase reaction and lowers the concentration of oxaloacetate, thus beginning another oscillation. Pyruvate kinase activity was made to oscillate by adding fructose bisphosphate (10 micromol/l). Fructose bisphosphate was degraded to triose phosphates fairly rapidly, and, as it was degraded, there was a parallel decrease in pyruvate kinase activity. The enzyme was reactivated and made to oscillate with subsequent additions of fructose bisphosphate. The mitochondrial glycerol phosphate dehydrogenase was made to oscillate by adding EGTA to chelate calcium, which activates the enzyme. When the concentration of free calcium was raised to >0.1 micromol/l by adding more calcium, the activity of the enzyme increased. Repeated additions of chelator and calcium caused the enzyme activity to oscillate. The results with these three enzymes and physiological concentrations of naturally occurring effectors raise the possibility that the activities of not only these enzymes but of numerous enzymes oscillate in vivo in response to levels of allosteric effectors and substrates. If this is the case, pacemaker activity may result from complex effects distributed across multiple regulatory sites in both the cytosol and mitochondria, rather than from a single enzyme acting as a primary pacemaker. PMID- 9392487 TI - Localization and functional role of synaptotagmin III in insulin secretory vesicles in pancreatic beta-cells. AB - Pancreatic beta-cells secrete insulin by Ca2+-triggered exocytosis of insulin containing large dense-core vesicles. Synaptotagmin is a Ca2+/phospholipid binding protein and is a good candidate for the Ca2+ sensor for exocytosis of synaptic vesicles in neurons. In the present study, we generated a polyclonal antibody against synaptotagmin III, and found that synaptotagmin III immunoreactivity was present at high levels in insulin-containing pancreatic islet cells and insulin-secreting clonal MIN6 cells. In subcellular fractionations of MIN6 cells, synaptotagmin III was recovered in the vesicular fractions containing both insulin and vesicle-associated membrane protein-2 (VAMP 2), but not in synaptophysin-positive fractions. The secretory vesicles immunoprecipitated by anti-VAMP-2 antibody contained synaptotagmin III and insulin. In addition, treatment of streptolysin-O-permeabilized MIN6 cells with anti-synaptotagmin III antibody significantly inhibited Ca2+-triggered insulin secretion. These results indicate that synaptotagmin III is localized in insulin containing dense-core vesicles in pancreatic beta-cells, and further strongly suggest that synaptotagmin III is the Ca2+ sensor in the exocytosis of insulin secretory vesicles. PMID- 9392488 TI - Assessment of hepatic sensitivity to glucagon in NIDDM: use as a tool to estimate the contribution of the indirect pathway to nocturnal glycogen synthesis. AB - NIDDM is associated with excessive rates of endogenous glucose production in both the postabsorptive and postprandial states. To determine whether this is due to an intrinsic increase in hepatic sensitivity to glucagon, 9 NIDDM and 10 nondiabetic subjects were studied on three occasions. On each occasion, glycogen was labeled the evening before the study with subjects ingesting meals containing [6-3H]galactose. Beginning at 6:00 A.M. on the following morning, somatostatin was infused to inhibit endogenous hormone secretion. Insulin concentrations were maintained constant at basal levels (defined as that necessary to keep glucose at approximately 5 mmol/l) in each individual. On one occasion, glucagon was infused at a rate of 0.65 ng x kg(-1) x min(-1) throughout the experiment, resulting in glucagon concentrations of approximately 130 pg/ml and a slow but comparable fall in endogenous glucose production with time in both groups. On the other two occasions, the glucagon infusion was increased at 10:00 A.M. to either 1.5 or 3.0 ng x kg(-1) x min(-1), resulting in an increase in glucagon concentrations to approximately 180 and 310 pg/ml, respectively. The increment in endogenous glucose production (i.e., area above basal) did not differ in diabetic and nondiabetic subjects during either the 1.5 ng x kg(-1) x min(-1) (0.75 +/- 0.055 vs. 0.78 +/- 0.048 mmol/kg) or 3.0 ng x kg(-1) x min(-1) (1.06 +/- 0.066 vs. 1.10 +/- 0.073 mmol/kg) glucagon infusions. In contrast, the amount of [6-3H]glucose released from glycogen was lower (P < 0.05) in the diabetic than nondiabetic subjects during both glucagon infusions. The specific activity of glycogen, calculated as the integrated release of [6-3H]glucose divided by the integrated release of unlabeled glucose, was lower (P < 0.05) in diabetic subjects than in nondiabetic subjects during both the 1.5 ng x kg(-1) x min(-1) (19.0 +/- 3.9 vs. 41.4 +/- 5.7 dpm/micromol) and 3.0 ng x kg(-1) x min(-1) (19.1 +/- 3.1 vs. 36.5 +/- 7.2 dpm/micromol) glucagon infusions, implying that a greater portion of the glucose released from glycogen was derived from the indirect pathway. We concluded that although NIDDM is not associated with an intrinsic alteration in hepatic sensitivity to glucagon, it does alter the relative contributions of the direct and indirect pathways to nocturnal glycogen synthesis. PMID- 9392489 TI - Effects of insulin on blood flow and volume in skeletal muscle of patients with IDDM: studies using [15O]H2O, [15O]CO, and positron emission tomography. AB - Exaggerated vasoconstriction and blunted vasodilation of peripheral resistance arteries to various vasoactive agents characterize patients with IDDM. We characterized the hemodynamic effects of insulin in skeletal muscle in patients with IDDM. Muscle blood flow and blood volume were measured basally and during a high-dose insulin infusion (5 mU x kg(-1) x min[-1]) in seven normotensive patients with IDDM (age, 30 +/- 6 years; BMI, 24.5 +/- 2.0 kg/m2; blood pressure, 124 +/- 12/78 +/- 11 mmHg) and nine matched normal subjects, using [15O]H2O, [15O]CO, and positron emission tomography (PET). Whole-body insulin sensitivity was determined using the euglycemic insulin clamp technique. Insulin-stimulated whole-body glucose uptake was significantly lower in the patients with IDDM (45 +/- 15 micromol x kg(-1) x min[-1]) than in the normal subjects (62 +/- 14 micromol x kg(-1) x min[-1]) (P < 0.05). Insulin increased muscle blood flow by 111 +/- 69% above basal from 3.0 +/- 2.0 to 5.8 +/- 3.0 ml x 100 g(-1) muscle x min(-1) (P < 0.005) in the normal subjects, but only by 42 +/- 30% from 2.0 +/- 0.9 to 2.9 +/- 1.4 ml x 100 g(-1) muscle x min(-1) (P < 0.005) in patients with IDDM (P < 0.05 for change in flow in IDDM vs. normal subjects). The calculated muscle vascular resistances were comparable basally, but higher during hyperinsulinemia in the patients with IDDM (37 +/- 17 mmHg x 100 g x min x ml[ 1]) than in the normal subjects (16 +/- 7 mmHg x 100 g x min x ml[-l]) (P < 0.05). Muscle blood volume increased significantly by insulin in both groups without any difference between the groups. We conclude that the ability of supraphysiological concentrations of insulin to stimulate muscle blood flow is blunted in patients with IDDM, because of the inability of insulin to stimulate linear flow velocity rather than blood volume in skeletal muscle. This defect adds yet another defect to the list of abnormalities in vascular function in IDDM, which might predispose these patients to develop hypertension. PMID- 9392490 TI - Diet-induced muscle insulin resistance in rats is ameliorated by acute dietary lipid withdrawal or a single bout of exercise: parallel relationship between insulin stimulation of glucose uptake and suppression of long-chain fatty acyl CoA. AB - Chronic high-fat feeding in rats induces profound whole-body insulin resistance, mainly due to effects in oxidative skeletal muscle. The mechanisms of this reaction remain unclear, but local lipid availability has been implicated. The aim of this study was to examine the influence of three short-term physiological manipulations intended to lower muscle lipid availability on insulin sensitivity in high-fat-fed rats. Adult male Wistar rats fed a high-fat diet for 3 weeks were divided into four groups the day before the study: one group was fed the normal daily high-fat meal (FM); another group was fed an isocaloric low-fat high glucose meal (GM); a third group was fasted overnight (NM); and a fourth group underwent a single bout of exercise (2-h swim), then were fed the normal high-fat meal (EX). In vivo insulin action was assessed using the hyperinsulinemic glucose clamp (plasma insulin 745 pmol/l, glucose 7.2 mmol/l). Prior exercise, a single low-fat meal, or fasting all significantly increased insulin-stimulated glucose utilization, estimated at either the whole-body level (P < 0.01 vs. FM) or in red quadriceps muscle (EX 18.2, GM 28.1, and NM 19.3 vs. FM 12.6 +/- 1.1 micromol x 100 g(-1) x min(-1); P < 0.05), as well as increased insulin suppressibility of muscle total long-chain fatty acyl-CoA (LC-CoA), the metabolically available form of fatty acid (EX 24.0, GM 15.5, and NM 30.6 vs. FM 45.4 nmol/g; P < 0.05). There was a strong inverse correlation between glucose uptake and LC-CoA in red quadriceps during the clamp (r = -0.7, P = 0.001). Muscle triglyceride was significantly reduced by short-term dietary lipid withdrawal (GM -22 and NM -24% vs. FM; P < 0.01), but not prior exercise. We concluded that muscle insulin resistance induced by high-fat feeding is readily ameliorated by three independent, short-term physiological manipulations. The data suggest that insulin resistance is an important factor in the elevated muscle lipid availability induced by chronic high-fat feeding. PMID- 9392491 TI - Leptin sensitivity in nonobese glucagon-like peptide I receptor -/- mice. AB - Glucagon-like peptide I (GLP-I) stimulates glucose-dependent insulin secretion and inhibits food intake in the central nervous system. Because leptin reduces food intake but inhibits insulin secretion, we examined leptin action in mice with a null mutation in the GLP-I receptor. Intracerebroventricular leptin administration inhibited food intake in both wild-type and GLP-I receptor (GLP IR) -/- mice, and daily intraperitoneal administration of leptin for 2 weeks produced comparable reductions in food intake and body weight in control and GLP IR -/- mice. Glucose tolerance was improved in both wild-type and GLP-IR -/- mice, whether pair fed or leptin treated; however, blood sugars were significantly lower in the leptin-treated GLP-IR -/- mice following oral glucose challenge (P < 0.01). Glucose-stimulated insulin was reduced in both pair-fed and leptin-treated mice (P < 0.01-0.001); however, insulin levels were significantly lower in leptin-treated versus pair-fed GLP-IR -/- mice (P < 0.01). A single leptin injection had no effect on glucose tolerance in GLP-IR -/- mice, but decreased hepatic PEPCK mRNA in both wild-type and GLP-IR -/- mice. The improvement in blood glucose excursion, despite lower levels of glucose stimulated insulin in lean leptin-treated GLP-IR -/- mice, suggests that leptin may have beneficial effects on control of blood glucose in the absence of obesity. Furthermore, the greater effects of leptin on glucose and insulin in leptin-treated versus pair-fed GLP-IR -/- mice raises the possibility that disruption of GLP-I signaling modifies the sensitivity to leptin in vivo. PMID- 9392492 TI - Leptin gene expression increases with age independent of increasing adiposity in rats. AB - Humans and rats tend to gain weight as they age. Leptin is one regulator of food intake and energy expenditure. To determine if the increase in adiposity with age is related to altered leptin gene expression, we assessed adiposity levels, leptin mRNA levels in epididymal and inguinal white adipose tissue (EWAT and IWAT), and uncoupling protein (UCP1) mRNA levels in interscapular brown adipose tissue (IBAT) from F344 x BN rats ages 3, 12, 18, 24, and 30 months (n = 8/age). Levels of adiposity determined by the adiposity index and the Lee index increased between ages 3 and 24 months, with a decrease at age 30 months. There were parallel increases with age in body weight, EWAT, and IWAT depot size up to age 24 months, followed by a nonsignificant decrease at age 30 months. Daily food intake was unchanged with age. In EWAT, leptin mRNA per microgram of RNA was unchanged with age, whereas in IWAT, it increased up to 24 months, then declined at 30 months. Total leptin mRNA levels in both IWAT and EWAT depots increased with age, peaking at age 24 months, and were correlated with adiposity. Serum leptin levels increased with age, also peaking at age 24 months, and were correlated with total leptin mRNA in WAT pads and adiposity. The rate of increase in serum leptin was greater than the increase in adiposity with age, suggesting contributions from both the increase in leptin expression per unit of WAT and the increase in WAT depot size. In addition, UCP1 mRNA levels in IBAT did not change with age. These data suggest that adiposity increases with age and cannot be attributed to increased food intake, impaired leptin gene expression, or decreased UCP1 mRNA level in IBAT. Furthermore, leptin gene expression in IWAT increases with age independent of increasing adiposity. PMID- 9392493 TI - Intracerebroventricular leptin increases lumbar and renal sympathetic nerve activity and blood pressure in normal rats. AB - Obesity and hyperinsulinism are known to be major stimuli of leptin production by adipose tissue, leading to increased leptin levels in the circulation. It has also been demonstrated that increased leptin production leads to satiety, possibly by decreasing the levels of neuropeptide Y (NPY) in the central nervous system (CNS). Because obesity and hyperinsulinism are also frequently associated with hypertension, we studied the effect of the intracerebroventricular (ICV) administration of leptin on mean arterial pressure (MAP), heart rate, vascular flows, and lumbar and renal sympathetic nerve activity (SNA). Normal Wistar rats were implanted with an ICV cannula and allowed to recover. On the day of the study, the animals were fasted and anesthetized with chloralose/urethane. Catheters were placed in a femoral artery and vein, and Doppler flow probes were placed around the iliac, renal, and superior mesenteric arteries for measurement of MAP, heart rate, and blood flows. In other experiments, lumbar SNA and renal SNA were recorded. ICV leptin administration resulted in an MAP that was slowly but progressively increasing. Blood flows decreased in the iliac and superior mesenteric arteries, but not in the renal artery. Leptin injection increased the lumbar SNA and renal SNA. The plasma glucose and insulin levels were not changed. We concluded that ICV leptin increases MAP by decreasing arterial blood flow to the skeletal muscle and the splanchnic vascular bed. This increased peripheral resistance is the result of an increased activity of the sympathetic nerves. We suggest that increased leptin may serve as a link in the triad of obesity and hyperinsulinism and hypertension. PMID- 9392494 TI - Method of insulin administration has no effect on insulin sensitivity estimates from the insulin-modified minimal model protocol. AB - The effect of the method of insulin administration on insulin sensitivity estimates from the insulin modified minimal model (MINMOD) protocol was evaluated using the tolbutamide-boosted protocol as a reference. The study included 21 nondiabetic men ages 40 +/- 2 years (mean +/- SE) with a BMI of 26.6 +/- 1.1 kg/m2. Each subject underwent four frequently sampled intravenous glucose tolerance tests (FSIGTT), one with tolbutamide and three with the same insulin dosage (0.03 U/kg) given as a bolus or infusion over 5 or 10 min. The insulin sensitivity index (SI) of each subject was calculated from each FSIGTT with MINMOD. Insulin sensitivity indexes from the four FSIGTTs were highly correlated (r > 0.85, P < 0.001). SI(insulin) from the bolus and the 5- and 10-min infusion protocols were similar, but were 21 +/- 5, 29 +/- 5, and 23 +/- 4% lower than SI(tolbutamide), respectively. SG(tolbutamide) and SG(insulin) were not different among the four protocols and were significantly correlated (r > 0.55, P < 0.01). Thus the tolbutamide and insulin protocols must not be used interchangeably in any single cross-sectional or longitudinal study. When the same insulin dosage is used, the method of its administration has no bearing on insulin sensitivity estimates from the insulin-modified FSIGTT. The same method of insulin administration should be used, however, in any single study for purpose of standardization. PMID- 9392496 TI - Impaired molecular regenerative responses in sensory neurones of diabetic rats: gene expression changes in dorsal root ganglia after sciatic nerve crush. AB - This study investigated changes in gene expression in lumbar dorsal root ganglia (DRG), contralateral and ipsilateral to a sciatic nerve crush in control and streptozotocin (STZ)-induced diabetic rats. After 10 weeks of diabetes, the left sciatic nerves of all rats were crushed at mid-thigh level, and the rats were maintained for a further 2 weeks. Northern blots, with internal standards, were made from L4 and L5 (pooled) DRG on each side to compare RNA hybrids from ganglia attached to crushed nerves with those attached to intact nerves. The expression of growth-associated proteins, GAP-43 and Talpha1 alpha-tubulin mRNA in DRG, was stimulated (all P < 0.05) by crush injury in control and diabetic rats. Steady state expression of transcripts for neurofilament (NF) proteins (NF-L, NF-H) and the high-affinity NGF receptor, trkA was decreased by diabetes in the contralateral ganglia to the crush (all P < 0.05). Crush injury further decreased expression of these transcripts in both control and diabetic rats (all P < 0.05). This reduced expression of mRNA coding for both growth-associated proteins, and neurofilament proteins in ganglia of diabetic rats could participate in the reduced competence of the regenerative response to nerve crush. PMID- 9392495 TI - Induction of renal kallikrein and renin gene expression by insulin and IGF-I in the diabetic rat. AB - The renal kallikrein-kinin system and the renin-angiotensin system are implicated in the pathogenesis of diabetic nephropathy. We have shown that renal kallikrein and renin gene expression are altered by diabetes. To investigate the cellular mechanisms responsible for these changes, we examined the effects of acute insulin and insulin-like growth factor I (IGF-I) treatment on renal kallikrein kinin and renin-angiotensin system components. Three weeks after induction of diabetes, we measured renal kallikrein and renin mRNA levels, renal kallikrein and renal renin activity, and plasma renin activity in control and diabetic rats and diabetic rats treated with insulin or IGF-I for 2 or 5 h. In diabetic rats, kallikrein and renin mRNA levels were reduced >50% compared with control rats. Renal tissue kallikrein levels and plasma renin activity were decreased, whereas renal renin content was unchanged. Insulin increased kallikrein and renin mRNA levels after 2 h. IGF-I, at a dosage that stimulated kallikrein mRNA levels in control rats, had no effect on renal kallikrein and renin content or mRNA levels in diabetic rats. However, infusion of a fivefold higher IGF-I dosage resulted in a two- to threefold increase in kallikrein and renin mRNA levels in 2 h. These data suggest that 1) diabetes suppresses kallikrein and renin gene expression, and these abnormalities are reversed by insulin or IGF-I; and 2) the diabetic state produces resistance to IGF-I induction of kallikrein and renin gene expression. These changes in regulated synthesis of kallikrein and renin in the kidney may underlie renal vascular changes that develop in diabetes. PMID- 9392497 TI - Hyperinsulinemia and abdominal obesity affect the expression of hypertriglyceridemia in heterozygous familial lipoprotein lipase deficiency. AB - We have reported three missense mutations (G188E, P207L, and D250N) in the lipoprotein lipase (LPL) gene among French-Canadians, resulting in the absence of measurable postheparin plasma LPL activity in homozygotes. Presence of triglyceride- and cholesterol-rich VLDL, as well as cholesterol-poor HDL particles, has been shown in heterozygotes affected by partial reduction in postheparin LPL activity. However, significant heterogeneity in their plasma triglyceride levels has been found, even among individuals carrying the same LPL gene mutation, indicating that factors other than LPL deficiency could affect the phenotypic expression of hypertriglyceridemia in the heterozygous state. The aim of the present study was to examine the combined effects of abdominal fat accumulation and hyperinsulinemia on plasma triglyceride levels among heterozygous patients for familial LPL deficiency. Based on sex and BMI, 43 heterozygotes (25 women and 18 men) were matched with noncarrier control subjects. Our data indicate that heterozygotes with higher abdominal fat deposition, as defined as waist girth values above the 50th percentile, had higher plasma triglyceride levels than nonobese heterozygotes. However, an important proportion of male heterozygote subjects were hypertriglyceridemic, even in absence of abdominal obesity, suggesting that another factor(s) was involved in the modulation of hypertriglyceridemia in these subjects. Indeed, multivariate analyses revealed that fasting hyperinsulinemia was a significant correlate of hypertriglyceridemia among these heterozygotes. Results of the present study indicate that abdominal obesity and hyperinsulinemia both have deleterious effects on plasma triglyceride levels in familial LPL deficiency. It is suggested that heterozygotes with moderate obesity and/or insulin resistance may be at higher risk of coronary artery disease because of the expression of an atherogenic lipoprotein phenotype among these patients. PMID- 9392498 TI - Altered platelet membrane dynamic properties in type 1 diabetes. AB - A modified platelet response to aggregating stimuli is supposed to play a role in the pathogenesis of diabetic macroangiopathy. We studied the fluidity and microheterogeneity of the external surface of the platelet membrane and the activities of the plasma membrane Na+-K+-ATPase and Ca2+-ATPase in 21 men with type 1 diabetes and in 20 control subjects before and after in vitro thrombin addition. In the resting state, platelets from type 1 diabetic patients showed an increased fluidity and microheterogeneity of the platelet membrane, a higher Ca2+ ATPase activity, and a reduced Na+-K+-ATPase activity in comparison with platelets from healthy subjects. The fatty acid composition was also modified, with increased C 16:1 and decreased C 18:0 content. Control cells incubated with thrombin showed a modification of the membrane parameters opposite to the response observed in type 1 cells after the stimulation. The incubation of control platelets in the resting state with high concentrations of glucose modified the fluidity of the plasma membrane Na+-K+-ATPase and Ca2+-ATPase activities in an opposite way in comparison with the alterations observed in type 1 platelets. This study suggests that in type 1 diabetic patients, the platelet membrane responds to activation with a molecular remodeling different from the response of healthy subjects. The abnormal organization of the membrane might contribute to the altered platelet functions in type 1 diabetic patients, but acute exposure to high glucose levels does not seem able to modify the platelet membrane in the way observed in type 1 diabetes. PMID- 9392499 TI - Increased expression of intercellular adhesion molecule-1 (ICAM-1) in diabetic rat glomeruli: glomerular hyperfiltration is a potential mechanism of ICAM-1 upregulation. AB - Mononuclear cells, including monocytes/macrophages and T-cells, are considered to be involved in the progression of diabetic nephropathy, although the mechanism of their recruitment into diabetic glomeruli is unclear. The intercellular adhesion molecule-1 (ICAM-1) promotes the infiltration of leukocytes into atherosclerotic lesions as well as inflammatory tissues. In the present study, we investigated the expression of ICAM-1 in the glomeruli of streptozotocin-induced diabetic rats. The expression of ICAM-1 was increased significantly during the early stage of diabetes. The number of mononuclear cells, primarily monocytes/macrophages and lymphocytes, was significantly increased in diabetic glomeruli. Mononuclear cell infiltration into diabetic glomeruli was prevented by anti-ICAM-1 monoclonal antibody. Insulin treatment decreased ICAM-1 expression and mononuclear cell infiltration. The ICAM-1 expression on cultured human umbilical vein endothelial cells was not induced under high glucose culture conditions. Glomerular hyperfiltration is a characteristic change in the early stage of diabetic nephropathy. Treatment with aldose reductase inhibitor, which prevented glomerular hyperfiltration without changes in blood glucose levels, decreased ICAM-1 expression and mononuclear cell infiltration. Moreover, we examined the ICAM-1 expression in the glomeruli of the 5/6 nephrectomized rat, which is a model for glomerular hyperfiltration without hyperglycemia. The ICAM-1 expression and infiltration of mononuclear cells was significantly increased in the glomeruli of 5/6 nephrectomized rats. We conclude that ICAM-1 is upregulated and promotes the recruitment of mononuclear cells in diabetic glomeruli. Moreover, glomerular hyperfiltration that occurs in the early stage of diabetic glomeruli may be one of the potential mechanisms of ICAM-1 upregulation in diabetic nephropathy. PMID- 9392500 TI - Cholesteryl ester transfer protein gene polymorphism is a determinant of HDL cholesterol and of the lipoprotein response to a lipid-lowering diet in type 1 diabetes. AB - The TaqIB cholesteryl ester transfer protein (CETP) gene polymorphism (B1B2) is a determinant of HDL cholesterol in nondiabetic populations. Remarkably, this gene effect appears to be modified by environmental factors. We evaluated the effect of this polymorphism on HDL cholesterol levels and on the lipoprotein response to a linoleic acid-enriched, low-cholesterol diet in patients with type 1 diabetes. In 44 consecutive type 1 diabetic patients (35 men), CETP polymorphism, apolipoprotein (apo) E genotype, serum lipoproteins, serum CETP activity (measured with an exogenous substrate assay, n = 30), clinical variables, and a diet history were documented. The 1-year response to diet was assessed in 14 type 1 diabetic patients, including 6 B1B1 and 6 B1B2 individuals. HDL cholesterol was higher in 10 B2B2 than in 14 B1B1 homozygotes (1.63 +/- 0.38 vs. 1.24 +/- 0.23 mmol/l, P < 0.01). HDL cholesterol, adjusted for triglycerides and smoking, was 0.19 mmol/l higher for each B2 allele present. CETP activity levels were not significantly different between CETP genotypes. Multiple regression analysis showed that VLDL + LDL cholesterol was associated with dietary polyunsaturated:saturated fatty acids ratio (P < 0.02) and total fat intake (P < 0.05) in the B1B1 homozygotes only and tended to be related to the presence of the apo E4 allele (P < 0.10). In response to diet, VLDL + LDL cholesterol fell (P < 0.05) and HDL cholesterol remained unchanged in 6 B1B1 homozygotes. In contrast, VLDL + LDL cholesterol was unaltered and HDL cholesterol decreased (P < 0.05) in 6 B1B2 heterozygotes (P < 0.05 for difference in change in VLDL + LDL/HDL cholesterol ratio). This difference in response was unrelated to the apo E genotype. Thus, the TaqIB CETP gene polymorphism is a strong determinant of HDL cholesterol in type 1 diabetes. This gene effect is unlikely to be explained by a major influence on the serum level of CETP activity, as an indirect measure of CETP mass. Our preliminary data suggest that this polymorphism may be a marker of the lipoprotein response to dietary intervention. PMID- 9392501 TI - Pyruvate improves deleterious effects of high glucose on activation of pentose phosphate pathway and glutathione redox cycle in endothelial cells. AB - In our previous study (Diabetes 44:520-526, 1995), endothelial cells cultured in high glucose condition showed impairment of an oxidant-induced activation of the pentose phosphate pathway (PPP) and a reduced supply of NADPH to the glutathione redox cycle. To gain insight into the mechanisms of this impairment, the protective effect of pyruvate was studied in human umbilical vein endothelial cells cultured in either 5.5 mmol/l glucose (normal glucose [NG] condition) or 33 mmol/l glucose (high glucose [HG] condition). Through pretreatment of cells with 0.2 mmol/l pyruvate for 5-7 days in the HG condition, glucose oxidation through the PPP and total cellular NADPH content in the presence of 0.2 mmol/l H2O2 were increased by 54 (P < 0.05) and 34%, respectively, and glutathione-dependent degradation of H2O2 in HG cells was enhanced by 41% (P < 0.01), when compared with those cells to which pyruvate was not added. The addition of pyruvate significantly reduced the fructose 1,6-bisphosphate (FDP) content and free cytoplasmic NADH/NAD ratio, estimated by increased pyruvate/lactate ratio in NG and HG cells exposed to H2O2. Furthermore, the addition of pyruvate also showed a 46% reduction (P < 0.01) of endothelial cell damage induced by H2O2 in HG cells. These results indicate that abnormalities in PPP activation and glutathione redox cycle activity induced by H2O2 in HG cells are compensated, and that the accentuated reductive stress is improved by an addition of pyruvate. These pyruvate effects are associated with protection against an oxidant-induced endothelial cell injury in the high glucose condition. PMID- 9392502 TI - Circulating vascular cell adhesion molecule-1 (VCAM-1) in atherosclerotic NIDDM patients. AB - Vascular cell adhesion molecule-1 (VCAM-1) has been shown to be highly expressed in atherosclerotic lesions. Although the soluble form of VCAM-1 (sVCAM-1) is detected in human sera, the relation between the degree of atherosclerosis and serum sVCAM-1 level has not been defined. In the present study, sVCAM-1 concentrations were measured in sera from 101 Japanese NIDDM patients. The mean +/- SD serum sVCAM-1 concentration in 26 patients with symptomatic atherosclerotic vascular diseases (789 +/- 187 ng/ml) was higher than that in 75 patients without the disease (664 +/- 175 ng/ml). Among the 101 NIDDM patients, 56 had atherosclerotic change of the carotid arteries, based on the evaluation by high-resolution B-mode ultrasonography. Their sVCAM-1 level was 759 +/- 201 ng/ml, higher than that in 45 patients without any detectable atherosclerosis of the carotid arteries (619 +/- 130 ng/ml). In addition, there was a positive correlation between sVCAM-1 concentration and thickness of the intimal plus medial complex (IMT) of the carotid arteries in the NIDDM patients (r = 0.41, P < 0.0001). Multivariate regression analysis revealed significant predictors of mean IMT value to be sVCAM-1 concentration (F = 62.88, P = 0.0001) and age (F = 9.59, P = 0.0026). By contrast, sVCAM-1 concentration was not increased in nondiabetic patients with atherosclerotic change of the carotid arteries (668 +/- 191 ng/ml; n = 36) compared with those without the atherosclerotic change (632 +/- 177 ng/ml; n = 28), and there was no correlation between sVCAM-1 level and IMT of the carotid arteries in the nondiabetic subjects. These results indicate that circulating sVCAM-1 may be a marker of atherosclerotic lesions in NIDDM patients with symptomatic and asymptomatic atherosclerosis. PMID- 9392503 TI - Association of methylenetetrahydrofolate reductase gene polymorphism with carotid arterial wall thickening and myocardial infarction risk in NIDDM. PMID- 9392504 TI - Uncoupling protein 2 region on chromosome 11q13 is not linked to markers of obesity in familial type 2 diabetes. PMID- 9392505 TI - An automated fluorescent single-strand conformation polymorphism technique for screening mutations in the hepatocyte nuclear factor-1alpha gene (maturity-onset diabetes of the young). PMID- 9392506 TI - Improved glucose tolerance restores insulin-stimulated Akt kinase activity and glucose transport in skeletal muscle from diabetic Goto-Kakizaki rats. AB - The serine/threonine kinase Akt (protein kinase B [PKB] or related to A and C protein kinase [RAC]) has recently been implicated to play a role in the signaling pathway to glucose transport. However, little is known concerning the regulation of Akt activity in insulin-sensitive tissues such as skeletal muscle. To explore the role of hyperglycemia on Akt kinase activity in skeletal muscle, normal Wistar rats or Goto-Kakizaki (GK) diabetic rats were treated with phlorizin. Phlorizin treatment normalized fasting blood glucose and significantly improved glucose tolerance (P < 0.001) in GK rats, whereas in Wistar rats, the compound had no effect on glucose homeostasis. In soleus muscle from GK rats, maximal insulin-stimulated (120 nmol/l) Akt kinase activity was reduced by 68% (P < 0.01) and glucose transport was decreased by 39% (P < 0.05), compared with Wistar rats. Importantly, the defects at the level of Akt kinase and glucose transport were completely restored by phlorizin treatment. There was no significant difference in Akt kinase protein expression among the three groups. At a submaximal insulin concentration (2.4 nmol/l), activity of Akt kinase and glucose transport were unaltered. In conclusion, improved glucose tolerance in diabetic GK rats by phlorizin treatment fully restored insulin-stimulated activity of Akt kinase and glucose transport. Thus, hyperglycemia may directly contribute to the development of muscle insulin resistance through alterations in insulin action on Akt kinase and glucose transport. PMID- 9392507 TI - Thiazolidinediones downregulate stearoyl-CoA desaturase 1 gene expression in 3T3 L1 adipocytes. AB - Thiazolidinediones (TZDs) are known to have potent increases of insulin sensitivity. Because peroxisome proliferator-activated receptor-gamma (PPAR gamma), a receptor for TZDs, is mainly expressed in adipocytes, we tried to search the TZD-targeted genes in mouse 3T3-L1 adipocytes. By the mRNA differential display method, one band repressed by troglitazone was obtained, which corresponded to the partial sequences of the stearoyl-CoA desaturase 1 (SCD1) gene. Troglitazone dramatically decreased SCD1 mRNA levels in 3T3-L1 adipocytes in a dose-dependent manner. Pioglitazone also repressed the SCD1 mRNA expression, whereas WY-14,643 had no apparent effect. Both troglitazone and pioglitazone raised the composition (weight percentage) of myristic acid (C14:0), palmitic acid (C16:0), and stearic acid (C18:0), but lowered the composition of the delta9-cis desaturated fatty acids such as myristoleic acid (C14:1, delta9), palmitoleic acid (C16:1, delta9), oleic acid (C18:1, delta9), and linoleic acid (C18:2, delta9,12). These results indicate that TZDs repress SCD1 activity in 3T3 L1 adipocytes via downregulating SCD1 enzyme gene expression. PMID- 9392508 TI - Leptin increases hypothalamic pro-opiomelanocortin mRNA expression in the rostral arcuate nucleus. AB - Melanocortins are peptides, cleaved from the pro-opiomelanocortin (POMC) precursor, that act in the brain to reduce food intake and are potential mediators of leptin action. In the forebrain, melanocortins are derived from POMC containing neurons of the hypothalamic arcuate nucleus. To test the hypothesis that these POMC neurons are regulated by leptin, we used in situ hybridization to determine whether reduced leptin signaling (as occurs in fasting), genetic leptin deficiency (in obese ob/ob mice), or genetic leptin resistance (in obese db/db mice) lower expression of POMC mRNA. We further hypothesized that leptin administration would raise hypothalamic POMC mRNA levels in leptin-deficient animals, but not in mice with defective leptin receptors. In wild-type mice (n = 12), fasting for 48 h lowered POMC mRNA levels in the rostral arcuate nucleus by 53%, relative to values in fed controls (n = 8; P < 0.001). Similarly, arcuate nucleus POMC mRNA levels were reduced by 46 and 70% in genetically obese ob/ob (n = 6) and db/db mice (n = 6), respectively, as compared with wild-type mice (n = 5) (P < 0.01 for both comparisons). Five daily intraperitoneal injections of recombinant murine leptin (150 microg) raised levels of POMC mRNA in the rostral arcuate nucleus of ob/ob mice (n = 8) by 73% over saline-treated ob/ob control values (n = 8; P < 0.01), but was without effect in db/db mice (n = 6). In normal rats, two injections of a low dose of leptin (3.5 microg) into the third cerebral ventricle (n = 15) during a 40-h period of fasting also increased POMC mRNA levels in the rostral arcuate nucleus to values 39% greater than those in vehicle treated controls (n = 14; P = 0.02). We conclude that reduced central nervous system leptin signaling owing to fasting or to genetic defects in leptin or its receptor lower POMC mRNA levels in the rostral arcuate nucleus. The finding that leptin reverses this effect in ob/ob, but not db/db, mice suggests that leptin stimulates arcuate nucleus POMC gene expression via a pathway involving leptin receptors. These findings support the hypothesis that leptin signaling in the brain involves activation of the hypothalamic melanocortin system. PMID- 9392509 TI - Harold Rifkin, MD. 1916-1997. PMID- 9392510 TI - Signalling networks regulating dental development. AB - There has been rapid progress recently in the identification of signalling pathways regulating tooth development. It has become apparent that signalling networks involved in Drosophila development and development of mammalian organs such as the limb are also used in tooth development. Teeth are epithelial appendages formed in the oral region of vertebrates and their early developmental anatomy resembles that of other appendages, such as hairs and glands. The neural crest origin of tooth mesenchyme has been confirmed and recent evidence suggests that specific combinations of homeobox genes expressed in the neural crest cells may regulate the types of teeth and their patterning. Signalling molecules in the Shh, FGF, BMP and Wnt families appear to regulate the early steps of tooth morphogenesis and some transcription factors associated with these pathways have been shown to be necessary for tooth development. Several of the conserved signals are also transiently expressed in the enamel knots in the dental epithelium. The enamel knots are associated with the characteristic epithelial folding morphogenesis which is responsible for the development of tooth shape and it is currently believed that the enamel knots function as signalling centres regulating tooth shape development. The developing tooth has proven to be an excellent model in studies of the molecular basis of patterning and morphogenesis of organs and it can be expected that continuing studies will rapidly increase the understanding of these mechanisms. PMID- 9392511 TI - Nerve-induced disruption and reformation of beta1-integrin aggregates during development of the neuromuscular junction. AB - The earliest biochemical change detected during synaptogenesis is a local elimination of muscle basal lamina proteins. To explore whether this provides signal(s) that regulate postsynaptic differentiation, we examined the effects of innervation on the distribution of beta1-integrins, which were initially present in scattered aggregates complexed with basal lamina ligands. These beta1-integrin aggregates disappear along paths of nerve contact as their basal lamina ligands are eliminated. New accumulations of these proteins then form during assembly of the postsynaptic apparatus. The new beta1-integrin aggregates at developing synapses form partly via a redistribution of mobile molecules on muscle surface. We thus consider whether (a) the removal of integrins' basal lamina ligands alters their cytoplasmic ligand-interactions, causing the dissociation of integrin clusters, and (b) this receptor modulation helps to transduce local changes in pericellular protease activity into cytoplasmic signals that control postsynaptic differentiation. PMID- 9392512 TI - Cloning and characterization of cDNAs encoding the integrin alpha2 and alpha3 subunits from Xenopus laevis. AB - Integrins containing the alpha2 and alpha3 subunits associate with the beta1 subunit to form distinct receptors with partially overlapping adhesive specificities. We report the cloning and sequence of cDNAs that encode the Xenopus orthologues of integrins alpha2 and alpha3 and the expression of these subunits during embryogenesis. Integrin alpha2 and alpha3 mRNAs are first expressed in the dorsal mesoderm and developing notochord at gastrulation. We also show that alpha3 mRNAs are expressed in the entire marginal zone of gastrulae dorsalized with LiCl but that this localization is lost in embryos ventralized by ultraviolet light. Immunoblots reveal that the alpha3 protein is expressed throughout early development, however, the alpha2 protein is not detected until late tailbud stages. Injection of full-length alpha3 transcripts into the animal poles of fertilized eggs results in embryonic defects in paraxial mesoderm attributed to the failure of somites to form segments. Injection of the alpha3 transcripts into the vegetal pole and overexpression of a 5'-truncated alpha3 control construct have no apparent affect on development or somite formation. These data suggest that normal position-specific expression of integrins is important in maintaining the proper organization of tissues during early amphibian morphogenesis. PMID- 9392513 TI - Regulation of the CRABP-I gene during mouse embryogenesis. AB - The cellular retinoic acid binding protein type I (CRABP-I) shows a highly specific expression pattern during mouse embryonic development. The tissues that express CRABP-I, i.e. the central nervous system (CNS), neural crest, branchial arches, limb bud and frontonasal mass, coincide with those that are most sensitive to unphysiological retinoic acid (RA) concentrations. We have investigated the transcriptional elements that are responsible for the spatiotemporal regulation of CRABP-I expression in the mouse embryo. We show here that a 16 kb fragment harbours all the elements needed for the correct spatiotemporal expression pattern. Upon further dissection of this fragment we have found that expression in the CNS is driven by elements in the upstream region of the gene, while expression in mesenchymal and neural crest tissue is regulated via element(s) located downstream of exon II of the gene. Two distinct fragments in the upstream region are required for expression in the CNS, as neither of these fragments alone is able to drive correct expression of a reporter gene in transgenic mice. DNAseI footprinting analysis of the two upstream fragments revealed the presence of a number of protected elements. One of these regulatory elements has the hallmarks of an RA response element, suggesting that CRABP-I expression in neural tissue can be directly modulated by RA via the RARs/RXRs. PMID- 9392514 TI - The SpHE gene is downregulated in sea urchin late blastulae despite persistence of multiple positive factors sufficient to activate its promoter. AB - Previous studies of the regulatory region of the SpHE (hatching enzyme) gene of the sea urchin Strongylocentrotus purpuratus (Wei, Z., Angerer, L.M., Gagnon, M.L. and Angerer, R.C. (1995) Characterization of the SpHE promoter that are spatially regulated along the animal-vegetal axis of the sea urchin embryo. Dev. Biol. 171, 195-211) have shown that approximately 330 bp is necessary and sufficient to promote high level expression in embryos of transgenes that reproduce the spatially asymmetric pattern of endogenous gene activity along the maternally determined animal-vegetal embryonic axis. Furthermore, SpHE regulatory elements appear to be redundant since several different combinations are sufficient to elicit strong promoter activity and many subsets function like the endogenous gene only in non-vegetal cells of the blastula (Wei, Z., Angerer, L.M. and Angerer, R.C. (1997) Multiple positive cis-elements regulate the asymmetric expression of the SpHE gene along the sea urchin embryo animal-vegetal axis. Dev. Biol., 187, 71-88). Here we demonstrate by in vivo footprinting that many cis elements on the endogenous promoter are occupied when the gene is active in early blastulae, but the binding of corresponding trans factors is significantly reduced when the gene becomes inactive in late blastulae. In addition, downregulation of the promoter is accompanied by a transition from a non nucleosomal to a nucleosome-like chromatin structure. Surprisingly, in vitro DNase I footprints of the 300 bp promoter using nuclear protein extracts from early and late blastulae are not detectably different and neither this sequence, nor a longer one extending to -1255, reproduces the loss of endogenous SpHE transcriptional activity after very early blastula stage. These observations imply that temporal repression of SpHE transcription involves a decrease in accessibility of the promoter to activators that are nevertheless present in nuclei and capable of activating transgene promoters. Temporal, but not spatial, downregulation is therefore likely to be regulated by negative activities functioning outside the -1255 promoter region which may serve as direct repressors or mediate an inactive chromatin structure. PMID- 9392515 TI - The characterization of novel Pax genes of the sea urchin and Drosophila reveal an ancient evolutionary origin of the Pax2/5/8 subfamily. AB - The developmental control genes of the Pax family can be grouped into different subclasses according to structure and sequence homology. Here we describe the isolation and characterization of three novel Pax genes of the sea urchin for which no homologues are yet known in other animal phyla. One of these genes, suPaxB, codes for the previously characterized transcription factor TSAP which is involved in the developmental regulation of two pairs of late histone genes. Furthermore, conserved members of the Pax2/5/8 subfamily, which have so far been described only in vertebrates, were isolated not only from the sea urchin, but also from Drosophila and C. elegans. Hence, the Pax2/5/8 transcription factors constitute an ancient subfamily of highly conserved Pax proteins. During Drosophila embryogenesis, the Pax258 gene is shown to be expressed in the precursor cells of the external sensory organs, thus suggesting a role for Pax258 in the early development of the peripheral nervous system of insects. PMID- 9392516 TI - Synergism between temporally distinct growth factors: bFGF, insulin and lens cell differentiation. AB - Fibroblast growth factors (FGFs) are the only known factors that can induce differentiation of the mammalian lens epithelial cell, while insulin acts only as a mitogen, not as a morphogen. We show here that insulin enhances expression of the alphaA-crystallin gene in lens epithelial cells and induces the synthesis of lens fibre cell specific betaB2- and gamma-crystallins in early differentiated fibre cells. Different signal transduction pathways are required for bFGF or insulin maintained fibre cell differentiation. A 15 min preincubation with bFGF was sufficient for the lens epithelial cells to become competent to undergo insulin maintained differentiation. The phorbol ester TPA could replace bFGF. The bFGF instructed competence to differentiate decays with a half-life of about 30 h. Hence, bFGF and insulin can act in concert to produce a differentiated phenotype even when they are not present simultaneously. PMID- 9392517 TI - Wing surface interactions in venation patterning in Drosophila. AB - The adult wing of Drosophila consists of two wing surfaces apposed by their basal membranes which first came into contact following disc eversion at metamorphosis. Veins appear in these surfaces in a dorsal-ventral symmetric pattern, but are 'corrugated' (vein cells are more compacted and more pigmented) in a dorsal ventral asymmetric pattern. We prevented dorsal-ventral contact apposition during wing imaginal disc morphogenesis by implanting fragments of discs into metamorphosing hosts. In these implants, longitudinal veins differentiate but with wider corrugation and in both surfaces. These results and those of genetic mosaics of mutants removing veins or causing ectopic veins reveal mutual dorso ventral induction/inhibition at work to modulate the final vein differentiation pattern and corrugation. PMID- 9392518 TI - Progesterone acts through protein kinase C to remodel the cytoplasm as the amphibian oocyte becomes the fertilization-competent egg. AB - The fertilization-competent Xenopus egg undergoes a contraction of its cortex towards the apex of the pigmented animal hemisphere within 10 min of fertilization. Evidence suggests that protein kinase C (PKC) is involved in the assembly of this contractile network and we show that PKC is rapidly activated as a result of exposure of oocytes to progesterone. Xenopus oocytes contain at least five different isotypes of PKC. Three actin-binding proteins (i.e. vinculin, talin and ankyrin) appear to play an early role in the assembly of the contractile network and one of the proteins (vinculin) becomes phosphorylated shortly after progesterone treatment as the contractile network is assembling. Our results indicated that progesterone acts through a phospholipase to activate PKC and that PKC participates in the remodeling of the cytoplasmic compartment as the oocyte becomes the egg. PMID- 9392519 TI - Analysis of a cDNA sequence encoding a novel member of the snake venom metalloproteinase, disintegrin-like, cysteine-rich (MDC) protein family from Agkistrodon contortrix laticinctus. AB - In this paper, we present a cDNA sequence encoding a full-length precursor form of a new member (ACLD) of the metalloproteinase-disintegrin-like protein family from the venom glands of Agkistrodon contortrix laticinctus (broad-banded copperhead) snake. Comparison of the deduced amino acid sequence of ACLD with those of other members of the metalloproteinase-disintegrin protein family from both mammalian and snake venom origin suggests that some conserved residues may be involved in processing of the disintegrin domain. PMID- 9392520 TI - Fungal riboflavin 5'-hydroxymethyl dehydrogenase catalyzes formation of both the aldehyde (riboflavinal) and the acid (riboflavinoic acid). AB - The purified enzyme from Schizophyllum commune that readily catalyzes the oxidation of the 5'-hydroxymethyl function of riboflavin (vitamin B2) with a redox dye and O2 to form the 5'-aldehyde can more slowly further oxidize the 5' aldehyde to the 5'-acid. Hence, the formation of these so-called 'schizoflavins' can be accounted for by the action of one enzyme. PMID- 9392521 TI - Psychrophilic enzymes: a thermodynamic challenge. AB - Psychrophilic microorganisms, hosts of permanently cold habitats, produce enzymes which are adapted to work at low temperatures. When compared to their mesophilic counterparts, these enzymes display a higher catalytic efficiency over a temperature range of roughly 0-30 degrees C and a high thermosensitivity. The molecular characteristics of cold enzymes originating from Antarctic bacteria have been approached through protein modelling and X-ray crystallography. The deduced three-dimensional structures of cold alpha-amylase, beta-lactamase, lipase and subtilisin have been compared to their mesophilic homologs. It appears that the molecular adaptation resides in a weakening of the intramolecular interactions, and in some cases in an increase of the interaction with the solvent, leading to more flexible molecular edifices capable of performing catalysis at a lower energy cost. PMID- 9392522 TI - Carboxyl terminal deletion analysis of tryptophan hydroxylase. AB - Tryptophan hydroxylase (TPH) catalyzes the rate-limiting step in the synthesis of serotonin and participates (in a non-rate-limiting fashion) in melatonin biosynthesis. In rabbit, TPH exists as a tetramer of four identical 51007 dalton (444 amino acids) protein subunits. An intersubunit binding domain responsible for tetramer formation of TPH was identified by assessing the role of a carboxyl terminal leucine heptad and 4-3 hydrophobic repeat. These repeats are conserved in all of the aromatic amino acid hydroxylases and have been shown to be required for the assembly of tyrosine hydroxylase tetramers. Polymerase chain reaction was utilized to create three TPH carboxyl terminal deletions (C delta8, C delta12 and C delta17) that sequentially remove members of the leucine heptad and 4-3 hydrophobic repeat. Each deletion and full-length recombinant TPH was expressed in bacteria to obtain soluble enzyme extracts for subsequent activity and structural analysis. It was found that removal of 8, 12 or 17 amino acids from the carboxyl terminus of TPH did not significantly alter enzymatic activity when compared to full-length recombinant TPH. However, the macromolecular structure of the deletions was dramatically affected as determined by dimeric and monomeric profiles on size exclusion chromatography. It can be concluded that amino acids 428-444 (the C-terminal 17 amino acids) comprise an intersubunit binding domain that is required for tetramer formation of TPH, but that tetramer assembly is not essential for full enzymatic activity. PMID- 9392523 TI - Adaptational changes in kinetic parameters of G6PDH but not of PGDH during contamination-induced carcinogenesis in livers of North Sea flatfish. AB - Kinetic parameters of glucose-6-phosphate dehydrogenase (G6PDH) and phosphogluconate dehydrogenase (PGDH) were determined in situ in livers of marine flatfish flounder that were caught in unpolluted areas in the open sea and in the highly polluted river Elbe (Germany). Analysis was performed quantitatively in liver sections using valid enzyme histochemical methods and image analysis. G6PDH but not PGDH was strongly affected by contaminant exposure and subsequent carcinogenesis. G6PDH showed a gradual decrease in Vmax and Km for glucose-6 phosphate in extralesional normal-looking liver tissue. Hepatocellular carcinomas also showed a low Km, whereas the Vmax was upregulated. These findings are interpreted as follows: prolonged challenges of the livers by pollutants inhibit or inactivate G6PDH and this is compensated for by reduction in Km. In carcinomas, G6PDH levels are upregulated but the low Km values are kept to increase the NADPH production capacity required in cancer cells showing that posttranslational regulation processes are important to control cellular metabolism under various environmental conditions. PMID- 9392524 TI - Further characterization of the two tetraheme cytochromes c3 from Desulfovibiro africanus: nucleotide sequences, EPR spectroscopy and biological activity. AB - The genes encoding the basic and acidic tetraheme cytochromes c3 from Desulfovibrio africanus have been sequenced. The corresponding amino acid sequences of the basic and acidic cytochromes c3 indicate that the mature proteins consist of a single polypeptide chain of 117 and 103 residues, respectively. Their molecular masses, 15102 and 13742 Da, respectively, determined by mass spectrometry, are in perfect agreement with those calculated from their amino acid sequences. Both D. africanus cytochromes c3 are synthesized as precursor proteins with signal peptides of 23 and 24 residues for the basic and acidic cytochromes, respectively. These cytochromes c3 exhibit the main structural features of the cytochrome c3 family and contain the 16 strictly conserved cysteine + histidine residues directly involved in the heme binding sites. The D. africanus acidic cytochrome c3 differs from all the other homologous cytochromes by its low content of basic residues and its distribution of charged residues in the amino acid sequence. The presence of four hemes per molecule was confirmed by EPR spectroscopy in both cytochromes c3. The g-value analysis suggests that in both cytochromes, the angle between imidazole planes of the axial histidine ligands is close to 90 degrees for one heme and much lower for the three others. Moreover, an unusually high exchange interaction (approximately 10[-2] cm[-1]) was evidenced between the highest potential heme ( 90 mV) and one of the low potential hemes in the basic cytochrome c3. The reactivity of D. africanus cytochromes c3 with heterologous [NiFe] and [Fe] hydrogenases was investigated. Only the basic one interacts with the two types of hydrogenase to achieve efficient electron transfer, whereas the acidic cytochrome c3 exchanges electrons specifically with the basic cytochrome c3. The difference in the specificity of the two D. africanus cytochromes c3 has been correlated with their highly different content of basic and acidic residues. PMID- 9392525 TI - Calcium binding to recoverin: implications for secondary structure and membrane association. AB - Recoverin is an EF-hand calcium-binding protein reportedly involved in the transduction of light by vertebrate photoreceptor cells. It also is an autoantigen in a cancer-associated degenerative disease of the retina. Measurements by circular dichroism presented here demonstrate that the binding of calcium to recoverin causes large structural changes. increasing the alpha helical content of the protein and decreasing its beta-turn, beta-sheet and 'other' structures. The maximum helical content (67%) was observed at 100 microM free calcium and, unlike calmodulin, decreased as the calcium concentration was modulated in either direction from this value. Fluorescence measurements indicated that recoverin may aggregate or undergo structural changes independent of calcium binding as the calcium concentration is increased above 100 microM. EGTA also appeared to affect the structure of recoverin independent of its chelation of calcium. While calcium-induced conformational changes have been proposed to alter the membrane binding of recoverin through association of its myristoylated amino terminus, in the experiments presented here the partitioning of recoverin between the cytoplasmic and membrane compartments of the rod photoreceptor outer segment was unaffected by the concentration of calcium, therefore it appears unlikely that a calcium-myristoyl switch acts alone to anchor recoverin directly to the membrane. These experiments were conducted with native recoverin which is heterogeneously acylated, but mass spectrometry confirmed that simple chromatographic methods could be devised to isolate the different forms of recoverin for further studies. PMID- 9392526 TI - Enzymatic characteristics of retinal dehydrogenase type I expressed in Escherichia coli. AB - We expressed RalDH(I) in Escherichia coli and have shown that it functions in vitro with the complex CRBP-retinal (cellular retinol-binding protein) as substrate, either generated in situ from the complex CRBP-retinol and microsomal retinol dehydrogenases or provided directly as CRBP-retinal. Recombinant RalDH(I) had kinetic constants with CRBP-retinal of: Hill coefficient 1.8; K0.5 0.8 microM; and Vm 1.5 nmol/min/mg of protein at 25 degrees C. Apo-CRBP inhibited the reaction with CRBP-retinal with an IC50 of 1.4 microM. Citral inhibited RalDH(I) with an IC50 of approximately 1 microM compared to an IC50 of approximately 12 microM for RalDH(II), but did not serve as substrate for RalDH(I). RalDH(I) did not catalyze efficiently the dehydrogenation of acetaldehyde, but showed higher Vmax/Km values for hexanal, octanal, decanal and benzaldehyde than for either propanal or retinal. These data extend the characterization of RalDH(I), show that apo-CRBP competes with holo-CRBP as substrate for RalDH(I), and expand insight into the pathways of retinoic acid biogenesis from the most abundant substrates in vivo, retinoid-liganded CRBP. PMID- 9392527 TI - New aspects on the kinetics of activation of ribosomal peptidyltransferase catalyzed peptide bond formation by monovalent ions and spermine. AB - The effect of NH4+ and K+ ions on the activity of ribosomal peptidyltransferase was investigated in a model system derived from Escherichia coli, in which AcPhe puromycin is produced by a pseudo-first-order reaction between the preformed AcPhe-tRNA-poly(U)-ribosome complex (complex C) and excess puromycin. Detailed kinetic analysis suggests that both NH4+ and K+ ions act as essential activators of peptidyltransferase by filling randomly, but not cooperatively, multiple sites on the ribosome. With respect to the NH4+ effect at 25 degrees C. the values of the molecular interaction coefficient (n), the dissociation constant (KA), and the apparent catalytic rate constant (kmax) of peptidyltransferase at saturating levels of NH4+ and puromycin are 1.99, 268.7 mM and 24.8 min(-1), respectively. The stimulation of peptidyltransferase by K+ ions at 25 degrees C (n = 4.38, KA = 95.5 mM, kmax = 9.6 min[-1]) is not as marked as that caused by NH4+ ions. Furthermore, it is evident that NH4+ at high concentration (200 mM) is effective in filling regulatory sites of complex C, which are responsible for the modulatory effect of spermine. The combination of NH4+ ions (200 mM) with spermine (300 microM) produces an additive increase in peptidyltransferase activity. Taken together, these findings suggest the involvement of two related pathways in the regulation of peptidyltransferase activity, one mediated by specific monovalent cations and the other mediated by spermine. PMID- 9392528 TI - Effect of genetic variation on the fatty acid-binding properties of human serum albumin and proalbumin. AB - In the circulation, non-esterified fatty acids are transported by albumin which also facilitates their removal from donor cells and uptake into receptor cells. We have studied whether genetic variations in the albumin molecule can affect its in vivo fatty acid-binding properties. The fatty acids bound to 25 structurally different variants and to their wildtype counterparts, isolated from heterozygous carriers, were determined gas chromatographically. The variants were proalbumins, albumins with single amino acid substitutions and glycosylated or truncated albumins. In eight cases the total amount bound to the variants was diminished (0.4-0.8-fold), and in seven cases the load was increased to 1.3 or more of normal. Twenty-one fatty acids were quantitated, and for 19 alloalbumins significant deviations from normal were found. Usually, changes in total and individual fatty acid binding were of the same type, but several exceptions to this rule was found. The glycosylated albumin Casebrook showed the largest changes, the total load and the amount of bound palmitate was 8.6 and 14 times, respectively, the normal. The most pronounced changes and the majority of cases of increased binding were caused by molecular changes in domain III. Mutations in domain I, II and the propeptide resulted in smaller effects, if any, and these were often reductions in binding. PMID- 9392529 TI - A new meiotic endonuclease from Coprinus meiocytes. AB - Two different types of Coprinus meiotic nuclease have been previously reported by the authors which are believed to be involved in meiotic chromosome recombination [1,2]. A third meiotic endonuclease was purified from the cap tissues of the basidiocarp of Coprinus cinereus. The enzyme is a 60 kDa molecule composed of a monopolypeptide as revealed by SDS-PAGE and FPLC-Sephacryl S-300 gel filtration. The enzyme belongs to a type of endonuclease which can preferentially digest single-stranded DNA and requires divalent cations as a co-factor, most commonly Mg2+ ions. In the presence of this co-factor, the enzyme converts the supercoiled plasmid DNA (form I) to both the relaxed form (form II) and the linear form (form III). Ca2+ ions can also function as a co-factor, though, in this case, not only is form I plasmid converted to form II, but a few ladder bands between form I and form II are also produced. The Ca2+ ion effect as a cofactor can be prevented with ATP. Immunohistochemical observation shows that the enzyme is distributed in the surface of the gills, which contain the meiotic tissues. These characteristics clearly differ from those of the meiotic nucleases reported previously. PMID- 9392530 TI - Different pathways for radiation-induced apoptosis. PMID- 9392531 TI - Long-term results of total lymphoid irradiation in the treatment of cardiac allograft rejection. AB - PURPOSE: To evaluate the short and long-term effects of total lymphoid irradiation (TLI) in the treatment of cardiac transplant rejection. METHODS AND MATERIALS: Between 1986 and 1995, 48 courses of TLI were delivered to 47 cardiac transplant patients. In 37 patients, TLI was administered for intractable allograft rejection despite conventional therapy while 10 patients received TLI prophylactically. The prescribed radiation dose was 8 Gy in 0.8 Gy fractions twice weekly to mantle and inverted-Y plus spleen fields. Postirradiation follow up ranged from 6 months to 9.1 years, with a mean of 3.1 years. RESULTS: The actual mean dose was 7.3 Gy delivered over a mean of 39 days. Fifty-six percent of patients required treatment delay or abbreviation because of thrombocytopenia, leukopenia, infection, or unrelated problems. In patients treated for intractable rejection, rejection rates dropped from 0.46 to 0.14 and to 0.06 episodes/patient/month before, during, and after TLI (p < 0.0001). Rejection rates continued to drop throughout follow-up. Prednisone requirements decreased from 0.41 mg/kg before treatment to 0.21 mg/kg afterward (p < 0.0001). The ratio of helper to cytotoxic-suppressor T-cells decreased during TLI from 1.33 to 0.89, and remained low at 0.44, 2-4 months after treatment. Infection rates were not increased and two patients developed malignancy. Rejection rates were high during prophylactic treatment and this protocol was abandoned. Three-year actuarial survival after irradiation was 60% for patients with intractable rejection and 70% for the prophylactic cohort. CONCLUSION: TLI is an effective treatment for control of intractable cardiac rejection. Episodes of rejection and steroid dosage requirements are decreased for up to 9.1 years. A possible mechanism of action is long term alteration in T-lymphocyte subsets. Patients experience transient bone marrow suppression but no increase in infection or bleeding. Long term complications of TLI are not appreciably different than conventional immunosuppression. PMID- 9392532 TI - Randomized trial comparing early postoperative irradiation vs. the use of nonsteroidal antiinflammatory drugs for prevention of heterotopic ossification following prosthetic total hip replacement. AB - PURPOSE: A randomized trial was undertaken to assess the comparative efficacy of early postoperative irradiation with either 5 or 7 Gy vs. the use of nonsteroidal antiinflammatory drug (NSAID) for prevention of heterotopic ossification (HO) following prosthetic total hip replacement (THP). METHODS AND MATERIALS: Between 1993 and 1994, 301 patients were randomized to receive postoperative irradiation (5 or 7 Gy) or NSAID. One hundred and thirteen patients were treated with NSAID (indomethacin 2 x 50 mg/day for 1 week), 93 patients were irradiated with a single 7 Gy fraction, 95 patients with a single 5 Gy fraction. The treatment volume included the soft tissues between the periacetabular region of pelvis and the intertrochanteric portion of the femur. X-rays of treated hips were obtained immediately and 6 months after surgery. Heterotopic ossification was scored according to the Brooker Grading system. One hundred patients receiving no prophylactic therapy after total hip arthroplasty between 1988 and 1992, were analyzed and defined as historical control group. RESULTS: Incidence of heterotopic ossification was 16.0% in NSAID-group (Brooker Score I: 8.0%; II: 6.2%; III: 1.8%; IV: 0%), 30.1% in 5 Gy group (Brooker Score I: 24.7%; II: 4.3%; III: 1.1%; IV: 0%), and 11.1% in 7 Gy group (Brooker Score I: 11.6%; II: 0%; III: 0%; IV: 0%). Regarding overall heterotopic ossification there was a significant difference between the NSAID group and the 5 Gy group (p < .015), respectively, between the 7 Gy group and the 5 Gy group (p < .0001). No significant difference was noted in the influence of overall HO between the NSAID and the 7 Gy group (p > 0.3). Analyzing the clinically significant HO (Brooker Score III and IV) patients irradiated with 7 Gy developed less HO than those treated with NSAID (p = 0.003). Incidence of HO was greater in the untreated historical control group (Brooker Score I: 26%; II: 15%; III: 19%; IV: 5%) than in all three prophylacticly treated groups. CONCLUSION: Prophylactic irradiation of the operative site after hip replacement with single a 7 Gy fraction is the most effective postoperative treatment schedule in prevention of clinically significant heterotopic ossification. This therapy modality is more effective than irradiation with a single 5 Gy fraction or use of NSAID. PMID- 9392533 TI - Brainstem tolerance to conformal radiotherapy of skull base tumors. AB - PURPOSE: The aim of this study was to analyze the long-term incidence of brainstem toxicity in patients treated for skull base tumors with high dose conformal radiotherapy. METHODS AND MATERIALS: Between 1974 and 1995, 367 patients with chordomas (n = 195) and chondrosarcomas (n = 172) of the base of skull have been treated with combined megavoltage photon and 160 MeV proton radiotherapy. Following 3D treatment planning with delineation of target volumes and critical nontarget structures dose distributions and dose-volume histograms were calculated. Radiotherapy was given an 1.8 Gy or CGE (=Cobalt Gray Equivalent) dose per fraction, with prescribed target doses ranging from 63 CGE to 79.2 CGE (mean = 67.8 CGE). Doses to the brainstem surface were limited to < or = 64 CGE and to the brainstem center to < or = 53 CGE. RESULTS: Follow-up time ranged from 6 months to 21.4 years (mean = 42.5 months). Brainstem toxicity was observed in 17 of 367 patients attributable to treatment, resulting in death of three patients. Actuarial rates of 5 and 10-year high-grade toxicity-free survival were 94 and 88%, respectively. Increased risk of brainstem toxicity was significantly associated with maximum dose to brainstem, volume of brainstem receiving > or = 50 CGE, > or = 55 CGE, and > or = 60 CGE, number of surgical procedures, and prevalence of diabetes or high blood pressure. Multivariate analysis identified three independent factors as important prognosticators: number of surgical procedures (p < 0.001), volume of the brainstem receiving 60 CGE (p < 0.001), and prevalence of diabetes (p < 0.01). CONCLUSIONS: Tolerance of brainstem to fractionated radiotherapy appears to be a steep function of tissue volume included in high dose regions rather than the maximum dose of brainstem alone. In addition, presence of predisposing factors as well as extent of surgical manipulation can significantly lower brainstem tolerance in the individual patient. PMID- 9392534 TI - The role of stereotactic radiosurgery in the treatment of malignant skull base tumors. AB - PURPOSE: To determine the efficacy and toxicity of stereotactic radiosurgery in the treatment of malignant skull base tumors. METHODS AND MATERIALS: Thirty-two patients with 35 newly diagnosed or recurrent malignant skull base tumors < or = 33.5 cm3 were treated using the Leksell Gamma unit. Tumor histologies included: adenoid cystic carcinoma [12], basal cell carcinoma [1], chondrosarcoma [1], chordoma [8], nasopharyngeal carcinoma [3], osteogenic sarcoma [2], and squamous cell carcinoma [8]. RESULTS: After a median follow-up of 2.3 years, 83% +/- 15% (+/-95% confidence interval) of patients experienced a symptomatic response to treatment. Local control at the skull base was 95 +/- 9% at 2 years and 78 +/- 23% at 3 years. Local-regional control above the clavicles was 75 +/- 15% at 1 year and 51 +/- 20% at 2 years. Overall and cause specific survival were identical, 82 +/- 13% at 1 year, 76 +/- 14% at 2 years, and 72 +/- 16% at 3 years. One patient developed a radiation-induced optic neuropathy 12 months after radiosurgery. CONCLUSION: Stereotactic radiosurgery using the Leksell Gamma Unit can provide durable tumor control and symptomatic relief with acceptable toxicity in the majority of patients with malignant tumors 4 cm or less in size involving the skull base. Further evaluation of more patients with longer follow-up is warranted. PMID- 9392535 TI - Analyses of neuro-otological complications after radiosurgery for acoustic neurinomas. AB - PURPOSE: To find out the optimum treatment parameters and the proper indications for treatment of acoustic neurinomas, univariate and multivariate actuarial analyses of neuro-otological complications after stereotactic radiosurgery for acoustic neurinomas were performed. METHODS AND MATERIALS: The subjects were 46 patients with acoustic neurinomas who underwent unilateral radiosurgery between June 1990 and June 1994 and were followed up at the University of Tokyo. Age ranged from 13 to 77 years (median, 54 years). Tumor diameter ranged from 0 to 25 mm (mean, 12 mm) at the cerebellopontine angle and from 2 to 15 mm (mean, 8.3 mm) in the internal auditory meatus. Maximum tumor doses ranged from 20 to 40 Gy (mean, 31.4 Gy), and peripheral doses from 12 to 25 Gy (mean, 16.8 Gy). One to eight isocenters were used (mean, 3.2). Median follow-up was 39 months. Eight events concerning neuro-otological complications were chosen, and the potential risk factors for them were analyzed by the actuarial analyses (univariate and multivariate). The events examined include hearing loss, vestibular function loss, facial palsy, and trigeminal nerve dysfunction. In order to point out potential risk factors for neuro-otological complications, univariate analyses were performed using both the Wilcoxon test and the log rank test, and multivariate analyses were performed with the Cox proportional hazards model. Variables nominated as potential risk factors were 1) demographic variables such as patient age and sex, 2) tumor dimensions, 3) treatment variables such as tumor doses and number of isocenters, and 4) pretreatment hearing levels. A variable with significant p-values (p < 0.05) in two or more of the three actuarial analyses (two univariate and one multivariate) was considered a possible risk factor. RESULTS: The possible variables that increase the risk for each event analyzed were: neurofibromatosis type II (NF2) and the number of isocenters for total hearing loss; experience of prior operation, the tumor diameter in the internal auditory meatus, and NF2 for hearing threshold elevation; peripheral tumor dose for vestibular function loss; patient age or midporus transverse tumor diameter (the two variables were correlated), and the number of isocenters for facial palsy; and the number of isocenters for trigeminal neuropathy. CONCLUSION: NF2 and the tumor diameter were the common risk factors for hearing loss in previous studies and ours. For the 5th/7th nerve dysfunction, the tumor diameter was the common risk factor. The risk of using more isocenters remains controversial. The difference in risk factors for hearing impairment and vestibular function loss suggests different mechanisms for the two. Further studies with larger populations and longer follow-up periods are required in order to draw conclusions on the risk factors in radiosurgery. PMID- 9392536 TI - 15 years experience with helium ion radiotherapy for uveal melanoma. AB - PURPOSE: To review the long-term experience of helium ion therapy as a therapeutic alternative to enucleation for uveal melanoma, particularly with respect to survival, local control, and morbidity. METHODS AND MATERIALS: 347 patients with uveal melanoma were treated with helium ion RT from 1978-1992. A nonrandomized dose-searching study was undertaken, with doses progressively reduced from 80 GyE in five fractions to 48 GyE in four fractions, given in 3-15 days, mean of 7 days. RESULTS: Local control was achieved in 96% of patients, with no difference in the rate of local control being seen at 80, 70, 60, or 50 GyE in five fractions. At the lowest dose level of 48 GyE in four fractions, the local control rate fell to 87%. Fifteen of 347 patients (4%) had local regrowth in the eye requiring enucleation (12 patients), laser (1 patient) or reirradiation (2 patients). The time of appearance of local regrowth ranged from 4 months to 5 years posttreatment, with 85% occurring within 3 years. Of the 347 patients, 208 are alive as of May 1, 1997. The median follow up of all patients is 8.5 years, range 1-17 years. Kaplan-Maier (K-M) survival is 80% at 5 years, 76% at 10 years, and 72% at 15 years posttreatment. Patients with tumors not involving the ciliary body have a 15-year K-M survival of 80%. The results for patients whose tumors involved the ciliary body are poor, with a 15-year K-M survival of 43%. Seventy-five percent of patients with tumors at least 3.0 mm from the fovea and optic nerve, and initial ultrasound height less than 6.0 mm, retained vision of 20/200 or better posttreatment. Patients with tumors larger than 6 mm in thickness, or with tumors lying close to the optic nerve or fovea, have a reduced chance of retaining useful vision. The enucleation rate is 19%, 3% for local failure and 16% because of complications of the helium RT, particularly neovascular glaucoma, which occurred in 35% of patients. CONCLUSIONS: Local control and retention of the eye are excellent. Complications of therapy reduce vision and eye preservation. Twenty-four percent of patients manifested distant metastases 6 to 146 months posttreatment, mean of 43 months, median of 36 months. Late-appearing distant metastases do not appear to be caused by persistent tumor in the eye. The risk of metastases is high for patients with tumors greater than 7 mm in initial ultrasound height (37%), anterior tumors involving the ciliary body (47%), and in those with local failure (53%). Patients with tumors not involving the ciliary body and initial dimensions less than 10 mm had only an 8% chance of death from melanoma. A search for effective adjuvant therapy is needed for patients at high risk of metastases (large tumors, ciliary body involved, local regrowth in eye). PMID- 9392537 TI - Anterior segment sparing to reduce charged particle radiotherapy complications in uveal melanoma. AB - PURPOSE: The purpose of this investigation is to delineate the risk factors in the development of neovascular glaucoma (NVG) after helium-ion irradiation of uveal melanoma patients and to propose treatment technique that may reduce this risk. METHODS AND MATERIALS: 347 uveal melanoma patients were treated with helium ions using a single-port treatment technique. Using univariate and multivariate statistics, the NVG complication rate was analyzed according to the percent of anterior chamber in the radiation field, tumor size, tumor location, sex, age, dose, and other risk factors. Several University of California San Francisco Lawrence Berkeley National Laboratory (LBNL) patients in each size category (medium, large, and extralarge) were retrospectively replanned using two ports instead of a single port. By using appropriate polar and azimuthal gaze angles or by treating patients with two ports, the maximum dose to the anterior segment of the eye can often be reduced. Although a larger volume of anterior chamber may receive a lower dose by using two ports than a single port treatment. We hypothesize that this could reduce the level of complications that result from the irradiation of the anterior chamber of the eye. Dose-volume histograms were calculated for the lens, and compared for the single and two-port techniques. RESULTS: NVG developed in 121 (35%) patients. The risk of NVG peaked between 1 and 2.5 years posttreatment. By univariate and multivariate analysis, the percent of lens in the field was strongly correlated with the development of NVG. Other contributing factors were tumor height, history of diabetes, and vitreous hemorrhage. Dose-volume histogram analysis of single-port vs. two-port techniques demonstrate that for some patients in the medium and large category tumor groups, a significant decrease in dose to the structures in the anterior segment of the eye could have been achieved with the use of two ports. CONCLUSION: The development of NVG after helium-ion irradiation is correlated to the amount of lens, anterior chamber in the treatment field, tumor height, proximity to the fovea, history of diabetes, and the development of vitreous hemorrhage. Although the influence of the higher LET deposition of helium-ions is unclear, this study suggests that by reducing the dose to the anterior segment of the eye may reduce the NVG complications. Based on this retrospective analysis of LBNL patients, we have implemented techniques to reduce the amount of the anterior segment receiving a high dose in our new series of patients treated with protons using the cyclotron at the UC Davis Crocker Nuclear Laboratory (CNL). PMID- 9392538 TI - External beam radiotherapy dose response of prostate cancer. AB - PURPOSE: To determine the external beam radiotherapy dose response of palpable Stage T1-T4, mostly Nx, patients with adenocarcinoma of the prostate. METHODS AND MATERIALS: There were 938 men consecutively treated between 1987 and 1995 who had pretreatment prostate specific antigen (PSA) levels. Posttreatment failure was defined as disease recurrence and/or two elevations in PSA on consecutive follow up visits. The radiotherapy technique consisted of a four-field box with a small four-field reduction after 46 Gy in 844 patients (total dose of 60-70 Gy) or with a six-field conformal boost after 46 Gy in 94 patients (total dose of 74-78 Gy). Neoadjuvant or adjuvant androgen ablation was not used in any patient. Median follow-up was 40 months. RESULTS: The mean and median radiotherapy doses for the entire group were 67.8 +/- 13.3 Gy (+/-SEM) and 66 Gy. The mean radiotherapy dose was higher in those who had Stage T3/T4 disease, Gleason scores of 8-10, or pretreatment PSAs of > 4 ng/ml. In general, patients with more aggressive pretreatment prognostic features were treated to higher doses; yet, those that relapsed or had a rising PSA were treated to significantly lower doses. Actuarial analyses were facilitated by dividing patients into three dose groups: < or = 67, > 67-77, and > 77 Gy. The actuarial freedom from failure rates at 3 years were 61, 74, and 96% for the low, intermediate, and high dose groups. Stratification of the patients by pretreatment PSA revealed that dose was a significant correlate of freedom from relapse or a rising PSA for those with PSAs > 4-10, > 10-20, and > 20 ng/ml. The only patients in which an improvement in outcome was not related to higher doses were those with a pretreatment PSA < or = 4 ng/ml. Dose was significantly associated with freedom from failure for Stage T1/T2 and Stage T3/T4 patients, as well as for those stratified by Gleason score. Multivariate analysis using Cox proportional hazards models showed that dose was an independent and highly significant predictor of relapse or a rising PSA. CONCLUSION: This retrospective review strongly indicates that radiotherapy dose to the prostate is critical to the cure of prostate cancer, even for favorable patients with pretreatment PSAs of > 4-10 ng/ml, Stages T1/T2, or Gleason scores of 2-6. Final confirmation awaits the results of our randomized trial. PMID- 9392539 TI - If you 'watch and wait,' prostate cancer may progress dramatically. AB - PURPOSE: Observation has been proposed as an option for localized prostate cancer. However, most series reporting on 'watch and wait' include patients treated by TUR or hormones that may affect results. We retrospectively reviewed the natural history of truly untreated prostate cancer and report the outcome for these patients. METHODS AND MATERIALS: From 1976 to 1992, 34 patients of median age 70 years (range 56-88) with biopsy proven localized adenocarcinoma of the prostate refused therapy. All had negative bone scan and none underwent TUR or hormone treatment. No patient was lost to follow-up (median 76 months). Failure patterns and survival were analyzed. RESULTS: At diagnosis 27 patients had palpable nodules (T2), of which 13 were well differentiated and 14 moderately differentiated. Seven had moderately differentiated T3 lesions. Mild prostatitis including nocturia, hesistancy, and urgency were reported in 16 T2 and 6 T3 patients. Within 36 months, local progression requiring therapy occurred in all T3, all T2 moderate and 5 of 13 T2 well-differentiated patients. Systemic progression occurred in 6 of 7 T3, 9 of 14 T2 (mod), and 2 of 13 T2 (well) patients. Overall 59% are alive, 26% succumbed to prostate carcinoma and 15% to other causes. CONCLUSION: Observation results in a high rate of local progression requiring intervention (77%) and excessive systemic disease development (50%) for patients with clinically palpable disease. Perhaps this strategy is viable for earlier stage lesions detected by PSA but it must be tested in a rigorous fashion before accepted. PMID- 9392540 TI - Prostate cancer patient subsets showing improved bNED control with adjuvant androgen deprivation. AB - PURPOSE: Cooperative groups have investigated the outcome of androgen deprivation therapy combined with radiation therapy in prostate cancer patients with variable pretreatment prognostic indicators. This report describes an objective means of selecting patients for adjuvant hormonal therapy by a retrospective matched case/control comparison of outcome between patients with specific pretreatment characteristics who receive adjuvant hormones (RT+H) vs. patients with identical pretreatment characteristics treated with radiation therapy alone (RT). In addition, this report shows the 5-year bNED control for patients selected by this method for RT+H vs. RT alone. METHODS AND MATERIALS: From 10/88 to 12/93, 517 T1 T3 NXM0 patients with known pretreatment PSA level were treated at Fox Chase Cancer Center. Four hundred fifty-nine of those patients were treated with RT alone while 58 were treated with RT+H. The patients were categorized according to putative prognostic factors indicative of bNED control, which include the palpation stage, Gleason score, and pretreatment PSA. We compared actuarial bNED control rates according to treatment group within each of the prognostic groups. In addition, we devised a retrospective matched case/control selection of RT patients for comparison with the RT+H group. Five-year bNED control was compared for the two treatment groups, excluding the best prognosis group, using 56 RT+H patients and 56 matched (by stage, grade, and pretreatment PSA level) controls randomly selected from the RT alone group. bNED control for the entire group of 517 patients was then analyzed multivariately using step-wise Cox regression to determine independent predictors of outcome. Covariates considered for entry into the model included stage (T1/T2AB vs. T2C/T3), grade (2-6 vs. 7-10), pretreatment PSA (0-15 vs. > 15), treatment (RT vs. RT+H), and center of prostate dose. bNED failure is defined as PSA > or = 1.5 ngm/ml and rising on two consecutive determinations. The median follow-up for the 112 matched case/control patients was 41 months. The median follow-up was 46 months for the RT (range 11-102 months) and 37 months for the RT+H group (range 6-82 months). RESULTS: Univariate analysis according to treatment for the prognostic factors of palpation stage, Gleason score, and pretreatment PSA demonstrates a significant improvement in 3 year bNED control with the addition of hormones for patients with T2C/T3, Gleason score 7-10, or pretreatment PSA > 15 ngm/ml. A comparison of bNED control according to treatment demonstrates improvement in 5-year bNED control of 55% for patients treated with RT+H vs. 31% for those patients treated with RT alone (p = 0.0088), although there is not a survival advantage. Multivariate analysis demonstrates that hormonal treatment is a highly significant independent predictor of bNED control (p = 0.0006) along with pretreatment PSA (p = 0.0001), palpation stage (p = 0.0001), grade (p = 0.0030), and dose (p = 0.0001). CONCLUSIONS: (1) Patients with specific adverse pretreatment prognostic factors (i.e., T2C/T3, Gleason score 7-10, pretreatment PSA > 15) benefit from adjuvant hormonal therapy. (2) Upon multivariate analysis, hormonal therapy is determined to be a highly significant predictor of bNED control, after adjusting for all other covariates. (3) The 5-year bNED control rates of 55% for RT+H vs. 31% for RT alone represents the magnitude of benefit from adjuvant hormone therapy. (4) The bNED control curves are separated by about 20 months, representing a delay in disease progression with adjuvant hormonal therapy, as there is no overall survival difference. PMID- 9392541 TI - Combined castration and fractionated radiotherapy in an experimental prostatic adenocarcinoma. AB - PURPOSE: The present study using the Dunning R3327-PAP rat prostatic adenocarcinoma model was designed to study the effect on tumor growth of castration prior to or after irradiation with 20-25 Gy as compared with either irradiation or castration alone. METHODS AND MATERIALS: Rats were bilaterally orchidectomized. During the irradiation procedure the nonanesthetized animals were held in a metallic frame with a strong cotton net and they were observed by means of a video camera. The suboptimal irradiation dose was given once daily with a 4-MeV linear accelerator, 4-5 Gy/fraction, during 5 consecutive days. Tumor volumes and rat weights were followed. At the end point of the study the animals were sacrificed and the tumors were morphometrically analyzed. RESULTS: The combination of irradiation and castration delayed tumor regrowth better than irradiation alone with the same suboptimal dose. Castration before irradiation delayed tumor regrowth more efficiently than castration after irradiation. However, castration alone delayed tumor regrowth even more effectively than suboptimal irradiation doses combined with castration. CONCLUSIONS: In combination with suboptimal irradiation neoadjuvant androgen deprivation was more inhibitory to rat prostatic adenocarcinoma regrowth than adjuvant androgen deprivation. Irradiation with suboptimal doses combined with castration may cause an earlier relapse to androgen-independent tumor growth than castration alone. PMID- 9392542 TI - Source localization following permanent transperineal prostate interstitial brachytherapy using magnetic resonance imaging. AB - PURPOSE: Dosimetric evaluation of completed brachytherapy implant procedures is crucial in developing proper technique and has prognostic implications. Accurate definition of the prostate gland and localization of the implanted radioactive sources are critical to attain meaningful dosimetric data. Methods using radiographs and CT accurately localize sources, but poorly delineate the prostate gland. MRI has been recognized as a superior imaging modality in delineating the prostate gland, but poor in localizing sources due to lack of source visibility. The purpose of this study was to optimize the visualization of sources using MRI and compare to CT derived source localization. METHODS AND MATERIALS: Multiple MRI scanning techniques were attempted until an acceptable sequence to visualize both the prostate gland and the implanted sources was found. The exams were performed using a pelvic coil only in approximately 15 min. The CT and MRI scans of 20 consecutive patients who had received TRUS-guided permanent transperineal interstitial prostate 125Iodine or 103Palladium brachytherapy were evaluated using an in-house dosimetry system. To eliminate anatomical dependence, the MRI derived DVHs for the entire calculation volume were then compared to those derived from the CT scans. RESULTS: The differences in isodose volumes, of the calculation volumes, for all implants at all dose levels were not statistically significant at the 95% confidence level. Calculation volume isodose volumes derived from MR images were statistically similar to those derived from CT images at the prescription dose for both 125Iodine (p < 0.01) and 103Palladium (p < 0.026). CONCLUSION: This study presents the first evidence that MRI may be reliably used to identify permanently implanted 125Iodine and 103Palladium sources. Given the advantage of target definition characteristics of MRI, substantially more accurate dosimetric analysis of prostate implants is now possible. The cost of the optimized and abbreviated MR scanning sequence used in this study is comparable to a pelvic CT scan. Postimplant MRI allows more accurate volumetric and anatomically relevant evaluation of permanent prostate implants, which may provide useful clinical correlation. PMID- 9392543 TI - Muscle invasive bladder cancer treated by transurethral resection, followed by external beam radiation and interstitial iridium-192. AB - PURPOSE: To evaluate the results of transurethral resection (TUR), external beam radiotherapy (EBRT), and interstitial radiation (IRT) with iridium-192, using the afterloading technique in patients with muscle invasive bladder cancer. METHODS AND MATERIALS: From May 1989 until September 1995, 66 patients with primary, solitary muscle invasive bladder cancer were treated with TUR, EBRT, and IRT, aiming at bladder preservation. According to the protocol, in three patients low dose EBRT was applied, whereas 63 patients received high-dose EBRT. Immediately prior to IRT, 42 patients underwent a lymphnode dissection, and in 16 cases a partial cystectomy was performed. For IRT, two to five catheters were used and IRT was started within 24 h after surgery. The majority of patients received 30 Gy of IRT, with a mean dose rate of .58 Gy/h. In three patients, additional EBRT was applied following IRT. Follow-up consisted of regular cystoscopies, mostly done during joint clinics of urologist and radiation oncologist, with urine cytology routinely performed. The median follow-up period was 26 months. The Kaplan-Meier method was used for the determination of survival rates. RESULTS: In seven patients, a bladder relapse developed. The probability of remaining bladder relapse free at 5 years was 88%. The bladder was preserved in 98% of the surviving patients. Metastases developed in 16 patients, and the probability of remaining metastasis free at 5 years was 66%. The cumulative 5-year overall and bladder and distant relapse free survival were 48% and 69%, respectively. Acute toxicity was not serious in the majority of cases; surgical correction of a persisting vesicocutaneous fistula was necessary in two patients, whereas a wound toilet had to be performed in another patient. Serious late toxicity (bladder, RTOG Grade 3) was experienced by only one patient. CONCLUSIONS: Interstitial radiation preceded by TUR and EBRT, in a selected group of patients with muscle invasive bladder cancer, yields an excellent bladder tumor control rate with a high probability of bladder preservation. Survival was mainly dependent on the development of distant metastases. Serious acute and late toxicity was rare. PMID- 9392544 TI - Radiosensitivity of Koch ileal reservoir. AB - PURPOSE: To acquire preliminary information on the radiosensitivity of the Koch ileal reservoir by reviewing acute and late toxicity incurred by nine patients who received pelvic radiotherapy after cystoprostatectomy with lower urinary reconstruction utilizing a Koch ileal reservoir with bilateral uretero-ileal urethrostomy. METHODS AND MATERIALS: All patients were irradiated because of synchronous locally advanced prostate cancer (pT3). A fourfield box technique at 100 cm source-axis-distance (SAD) with all fields treated every day at 1.8 Gy daily fractions, to a total dose of 45-50.40 Gy was used. The average AP portal dimension was 11 x 11 cm, and the average lateral was 7 x 8 cm. All portals were shaped using custom shields to optimize protection of normal tissues not suspected of tumor involvement (small bowel, posterior rectal wall). No attempt was made to shield the Koch ileal reservoir. For each patient, comparison of the treatment portals with the Kochgram radiography (gravity cystogram) confirmed the inclusion of the majority of the Koch ileal reservoir within the radiation fields. Acute and late morbidity was measured by RTOG toxicity criteria by retrospectively reviewing the patients' records. RESULTS: Only mild acute toxicity was reported by the patients: Six patients experienced grade 1 acute urinary toxicity and one suffered Grade 2 acute urinary toxicity. In four patients Grade 1 acute gastrointestinal toxicity occurred and in two patients Grade 2 toxicity occurred. With a median follow-up of 50 months late toxicity consisted mainly of microscopic hematuria in six patients and persistent frequency in two patients (with spontaneous improvement respectively at 4 and 6 months after radiation). No patients experienced acute or late Grade 3 or 4 genitourinary or gastrointestinal toxicity. CONCLUSION: The use of moderate doses of pelvic radiotherapy (45-50.40 Gy) at standard fractionation was well tolerated among nine patients who received pelvic radiation for invasive prostate cancer detected at the time of cystectomy and Koch ileal reservoir diversion. These preliminary data support the evidence that patients with a Koch ileal reservoir could safely undergo postoperative pelvic radiotherapy in these dose ranges and fractionation. PMID- 9392545 TI - Original p53 status predicts for pathological response in locally advanced breast cancer patients treated preoperatively with continuous infusion 5-fluorouracil and radiation therapy. AB - PURPOSE/OBJECTIVE: 1) To test feasibility of preoperative continuous infusion (c.i.) 5-Fluorouracil (5-FU) and radiation (RT) in locally advanced breast cancer. 2) To study clinical and pathological response rates of 5-FU and radiation. 3) To attempt preliminary correlations between biological probes and pathological response. METHODS AND MATERIALS: Previously untreated, locally advanced breast cancer patients were eligible: only patients who presented with T3/T4 tumors that could not be resected with primary wound closure were eligible, while inflammatory breast cancer patients were excluded. The protocol consisted of preoperative c.i. infusion 5-FU, 200 mg/m2/day with radiotherapy, 50 Gy at 2 Gy fractions to the breast and regional nodes. At mastectomy, pathological findings were classified based on persistence of invasive cancer: pathological complete response (pCR) = no residual invasive cells in the breast and axillary contents; pathological partial response (pPR) = presence of microscopic foci of invasive cells in either the breast or nodal specimens; no pathological response (pNR) = pathological persistence of tumor. For each patient pretreatment breast cancer biopsies were analyzed by immunohistochemistry for nuclear grade, ER/PR hormonal receptors, her2/neu and p53 overexpression. RESULTS: Thirty-five women have completed the protocol and are available for analysis. 5-FU was interrupted during radiation in 10 of 35 patients because of oral mucositis in 8 patients, cellulitis in 1, and patient choice in another. Objective clinical response rate before mastectomy was 71% (25 of 35 patients): 4 CR, 21 PR. However, in all 35 patients tumor response was sufficient to make them resectable with primary wound closure. Accordingly, all patients underwent modified radical mastectomy: primary wound closure was achieved in all patients. At mastectomy there were 7 pCR (20%), 5 pPR (14%) and the remaining 23 patients (66%) had pathological persistence of cancer (pNR). Variables analyzed as potential predictors for pathological response (pPR and pCR) were: initial TNM clinical stage, clinical response, nuclear grade, hormonal receptor status, p53 overexpression, and Her2/neu overexpression in the pretreatment tumor biopsy. Only initial p53 status (lack of overexpression at immunohistochemistry) significantly correlated with achievement of a pathological response to this regimen (p = 0.010). CONCLUSION: The combination of c.i. 5-FU and radiation was well tolerated and generated objective clinical responses in 71% of the patients. With the limitation of the small sample size, the complete pathological response achieved (20%) compares favorably with that reported in other series of neoadjuvant therapy for similar stage breast cancer. These preliminary data suggest that initial p53 status predicts for pathological response (pPR and pCR) to the combination of c.i. 5-FU and radiotherapy in locally advanced breast cancer. PMID- 9392546 TI - The role of regional nodal irradiation in the management of patients with early stage breast cancer treated with breast-conserving therapy. AB - PURPOSE: To determine the incidence of regional nodal failure (RNF) and indications for regional nodal irradiation (RNI) in patients with Stage I and II breast cancer treated with breast-conserving therapy (BCT). METHODS AND MATERIALS: Four hundred fifty-six patients with Stage I/II breast cancer were treated with BCT at William Beaumont Hospital. All patients underwent excisional biopsy and 288 (63%) were reexcised. A Level I/II ipsilateral axillary lymph node dissection was performed on 431 patients (95%). Pathologically involved nodes were found in 106 (23%) cases (69 with one to three nodes and 37 with > or = four nodes involved). All patients received whole breast irradiation (median dose 50 Gy) and 415 (91%) were boosted to the tumor bed (median total dose 60.4 Gy). Three hundred and sixty (79%) patients received breast alone irradiation and 96 (21%) also received RNI. The median axilla/supraclavicular fossa dose was 50 Gy. RESULTS: With a median follow-up of 83 months, 15 patients developed a RNF for a 5- and 8-year actuarial rate of 3 and 4%, respectively. The 5- and 8-year actuarial rates of axillary failure (AF) were 0.7 and 1.0%, respectively. The incidence of RNF or AF was not affected by the use of RNI in N0 or N1 patients with one to three positive nodes. Only in patients with four or more positive nodes was there a trend towards improved regional control with RNI (p = 0.09). However, patient numbers were extremely small, and this improvement was limited to a reduction in the rate of failure in the supraclavicular fossa (SCF) (20 vs. 0%, p = 0.04). Multiple clinical, pathologic, and treatment related factors were analyzed for an association with AF. On univariate analysis, AF was associated with the number of lymph nodes excised (p < 0.0001) estrogen receptor status (p = 0.0016), and pathologic node status (p = 0.0021). CONCLUSIONS: Regional nodal failure as the first site of failure is uncommon in patients with early-stage breast cancer treated with BCT with < or = three positive lymph nodes and appears unaffected by RNI. For patients with four or more positive lymph nodes, a trend towards improved RNF was noted with RNI, primarily in the SCF. However, patient numbers were extremely small in all subsets analyzed. Additional studies are needed to further define the need for RNI in these patients and help determine other factors associated with RNF. PMID- 9392547 TI - Influence of prognostic groupings and treatment results in the management of unresectable hepatoma: experience with Cisplatinum-based chemoradiotherapy in 76 patients. AB - PURPOSE: Internationally, hepatoma is a common cause of cancer death. Although the only curative therapy is surgical, most tumors are unresectable and cause death. The value of nonsurgical, antineoplastic therapy for such tumors is controversial. This study was undertaken to extend and confirm promising, but preliminary, treatment observations in the unresectable context. METHODS AND MATERIALS: From 1988 to 1993, 76 patients with unresectable, biopsy proven, hepatoma underwent uniform pretreatment assessment followed by induction therapy with external beam radiotherapy (21 Gy/7 fractions/10 days) and intravenous Cisplatinum, 50 mg/m2. One month later patients began monthly intrahepatic artery Cisplatinum, 50 mg/m2. Clinical course and treatment outcomes were correlated with previously published prognostic factors and groupings (Nomura et al., Okuda et al., Stillwagon, et al.). RESULTS: The toxicity of this therapy was modest and nonlimiting. Twenty-four patients (32%) progressed during induction and prior to receiving two cycles of intrahepatic artery Cisplatinum without evidence of benefit. Patients showing this early progression were more likely to be Stillwagon unfavorable than favorable (p = 0.013), Okuda Stage II than Stage I (p = 0.024), and slightly but not statistically more likely to be alpha-fetoprotein positive than alpha-fetoprotein negative (p = 0.098). The overall objective response rate was 43% (38% among AFP positive and 62% among AFP negative patients) (p = 0.15). Although 21 patients had evidence of extra hepatic metastases, survival for these patients did not differ from patients without metastases (p = 0.09) and patients with extra hepatic metastases were just as likely to show intrahepatic response (p = 0.84). CONCLUSION: The chemoradiotherapy program utilized produced objective response and minimal toxicity. One-third of patients progressed rapidly in spite of treatment. Among the remaining patients, response occurred frequently. This treatment appears to represent an important therapeutic option for many, but not all, patients with unresectable hepatoma. PMID- 9392548 TI - A phase I trial of hepatic arterial bromodeoxyuridine and conformal radiation therapy for patients with primary hepatobiliary cancers or colorectal liver metastases. AB - PURPOSE: We have previously found that conformal radiation therapy (RT) and hepatic arterial fluorodeoxyuridine was associated with durable responses and long-term survival for patients treated for nondiffuse primary hepatobiliary tumors and colorectal liver metastases. Further improvements in hepatic control may result from the addition of selective radiosensitization using bromodeoxyuridine (BrdU) infused through the hepatic artery (HA) concurrently with RT. This is a Phase I study of escalating doses of HA BrdU combined with our standard hepatic RT. METHODS AND MATERIALS: Patients with unresectable primary hepatobiliary cancer or colorectal liver metastases were treated with concurrent HA BrdU and conformal RT (1.5 Gy per fraction, twice a day). Three-dimensional treatment planning was used to define both the target and normal liver volumes. The total dose of RT (24, 48, or 66 Gy) was determined by the fractional volume of normal liver excluded from the high dose volume. HA BrdU was escalated in standard Phase I fashion with at least three patients receiving each combination of RT dose and BrdU dose. The starting dose of HA BrdU was 10 mg/kg/day, with two potential escalations to a maximum of 25 mg/kg/day (the maximum tolerable dose of HA BrdU when given alone on this same schedule). Grade > or = 3 toxicity was considered dose limiting. Patients receiving 24 Gy had one cycle of HA BrdU, while those receiving either 48 or 66 Gy had two cycles. Patients were followed for toxicity, complications, and response (when evaluable). RESULTS: A total of 41 patients (18 with colorectal liver metastases, 16 with cholangiocarcinoma and 7 with hepatoma) were treated. Five patients were removed from the protocol (three had HA catheter complications, one developed atrial fibrillation, and one was removed due to recurrent Grade 4 toxicity), although all five are included for toxicity purposes. Dose-limiting toxicity was primarily thrombocytopenia and there was no obvious relationship with the RT dose. Only 2 of 17 cycles given at 25 mg/kg/day had Grade > or = 3 toxicity. Complications developed in four patients, including one patient with radiation-induced liver disease. Response rates were not improved compared to our previous experience. CONCLUSIONS: The appropriate dose of HA BrdU for Phase II evaluation is 25 mg/kg/day. Neither the hepatic parenchyma nor the gastrointestinal mucosa appeared to be sensitized by this method of BrdU administration. It is anticipated that these, or still newer methods of therapy, can improve treatment results in the near future. PMID- 9392549 TI - Intraoperative radiation therapy combined with limited lymph node resection in gastric cancer: an alternative to extended dissection? AB - PURPOSE: To describe the results of a series of 63 Western patients presenting with gastric adenocarcinoma and treated with surgery and intraoperative radiation therapy (IORT) over a 8-year period and to discuss the role of IORT when combined with limited lymph node dissection. METHODS AND MATERIALS: From 1986 to 1993, 63 patients with gastric adenocarcinoma have been operated in the department of radiation oncology of the Hospices Civils de Lyon. The stage was: I in 17, II in 11, IIIA in 9, IIIB in 20, and IV in 6. The lymph node dissection was considered to be limited in 56 patients and extended in 7. The IORT dose ranged from 12 to 23 Gy (median: 15). Thirty patients also underwent a postoperative external beam irradiation with a standard dose of 44-46 Gy. RESULTS: The postoperative mortality rate was 4.8%. The 5-year overall survival in the entire series was 47% and was 82, 55, 78, 20, and 0% in Stages I, II, IIIA, IIIB, and IV, respectively. Loco-regional relapse occurred in 15 of 63 patients and metastases in 15 of 63. CONCLUSION: In Western patients treated by gastrectomy for adenocarcinoma of the stomach, IORT combined with limited lymph node dissection may provide overall survival similar to that observed after gastrectomy with extended lymph node dissection but with less postoperative mortality. PMID- 9392550 TI - Impact of clinical and therapeutic factors on major late complications after radiotherapy with or without concomitant chemotherapy for anal carcinoma. AB - PURPOSE: To investigate factors potentially influencing major late morbidity after sphincter-conserving treatment for anal carcinoma. METHODS AND MATERIALS: Grade 3-4 complications were retrospectively analyzed in 144 evaluable patients (pts), 55 pts after split-course radiotherapy (RT), and 89 after concomitant chemo-RT. First sequence RT delivered a median dose of 39.6 Gy using megavoltage photon beams. Boost treatment used either 192Ir implantation or external beam RT (median dose 20 Gy). Chemotherapy started on day 1 and in 83% of pts consisted of Mitomycin-C (10 mg/m2) and a 5-day infusion of 5-fluorourcil (600-800 mg/m2/day). Uni- and multivariate analyses tested the association of following factors with complication rate: age, gender, stage, anatomic tumor extent, type of biopsy, external RT technique (dose, fraction size, field arrangement), boost type (brachytherapy vs. external), brachytherapy dose and dose rate, overall treatment time, and addition of chemotherapy. RESULTS: Five-year actuarial complication rate was 16%. Two variables were significantly associated with complication rate: anatomic tumor extent (canal or margin vs. both +/- rectum; 10 vs. 31% complications, p = 0.0004) and first sequence prescribed dose (< 39.6 Gy vs. > or = 39.6 Gy; 7 vs. 23% complications, p = 0.012), confirmed as independent factors by Cox analysis. Grade 4 anal morbidity correlated significantly with prior local excision. All six bone complications were observed in pts treated by chemo-RT using large pelvic fields, five occurring in pts older than 66. CONCLUSION: Pts with tumors involving more than one anatomic subsite or treated with the higher first sequence RT dose are at greater risk of major complications. Prior tumor excision and combined modality therapy in older pts appear to favor major anal and bone complications, respectively. PMID- 9392551 TI - A phase I/II study of paclitaxel (TAXOL) and concurrent radiotherapy in advanced nonsmall cell lung cancer. AB - PURPOSE: The addition of chemotherapy to radical radiotherapy (XRT) has been shown to improve survival in locally advanced nonsmall cell lung cancer (9). Consequently, different chemotherapeutic regimens in combination with XRT are being evaluated in the treatment of this disease. Paclitaxel (TAXOL) may be a valuable drug in this situation as, in addition to a demonstrated activity in NSCLC, it has been shown to enhance the effect of radiation on cell lines in vitro. METHODS AND MATERIALS: Seventeen patients were enrolled onto a Phase I/II trial to determine the maximum tolerated dose of paclitaxel given by a 3-h infusion every 2 weeks throughout a 6-week course of XRT, 60 Gy in 30 daily fractions, in patients with Stage III NSCLC and then to describe the response rate of this combination in an expanded cohort of patients treated at the recommended phase II dose. Three patients were entered at each dose level (45, 90, 120, and 135 mg/m2), except for the 120 mg/m2 dose level, which was expanded to nine patients. RESULTS: The dose limiting toxicity was neutropenia--two of three patients treated at the 135 mg/m2 level experienced Grade 3 neutropenia on day 15, which precluded administration of scheduled chemotherapy. Esophagitis was mild to moderate, and although profound lymphopenia was observed at all dose levels, there was no evidence of associated opportunistic infections. Of the nine patients treated at the recommended Phase II dose of 120 mg/m2, there were one complete and six partial responses (response rate 78%). CONCLUSION: The combination of XRT, 60 Gy in 6 weeks and paclitaxel, 120 mg/m2 q 2 weeks, can be safely given to patients with NSCLC, and although it demonstrates activity in this situation, consideration should be given to the addition of other agents, such as platinum compounds. PMID- 9392552 TI - High dose rate brachytherapy for carcinoma of the oral tongue. AB - PURPOSE: The purpose of this study is to assess the feasibility of treating early staged tongue cancer with high dose rate (HDR) remote afterloading technique. Furthermore, a new figure of merit, the Geometry Index (GI), is introduced to quantify the quality of the implants. METHODS AND MATERIALS: Between 1994 and 1995, eight patients with carcinoma of the oral tongue were treated solely with interstitial implant using the HDR remote afterloading technique. Five patients had T1 N0 disease and the remaining three had T2 N0 disease. Elective neck treatment was withheld. The male-to-female ratio was 1:1, and the mean age 60 years (range: 32-72 years). The median follow-up time was 26 months (range: 6-30 months). The afterloading catheters were positioned through the submandibular approach with the assistance of templates. Six patients had single planar implant and the remaining two had double planar implant. The median number of catheters inserted was 5 (range: 4-9). The median dose given was 60 Gy in 10 fractions over 6 days. The interfraction interval was 7 h. Mandibular and maxillary shields were inserted prior to treatment. Thomadsen et al. introduced the use of Implant Quality Index (QI). We introduce a new parameter, GI, which is defined as ratio of the QI of the nonoptimized executed implant to the corresponding QI value of the nonoptimized idealized implant. RESULTS: The mucositis lasted for 6 to 20 weeks (median: 10 weeks). There was no local failure up to a median follow-up of 26 months. Two patients developed ipsilateral neck node metastases at 2 and 4 months following implant, respectively. One patient had involvement at level II and the other failed at level I to III. Both patients were salvaged by neck node dissection and regionally remained in control. One patient with multiple nodal metastases and extracapsular spread developed biopsy-proven liver metastases and succumbed 6 months following implant. One patient treated with double planar implant developed Grade 3 necrosis of the soft tissue and bone. This complication is largely preventable now, as we have acquired more technical expertise. The mean GI values for the single and double planar implants were 0.88 (range: 0.84 0.91) and 0.8, respectively. This correlates with our practical experience that it is more difficult to maintain a good geometry as double planar implant is required. The GI gives a better view of the geometry of implant as it compares the nonoptimized QI of the executed implant with its ideal counterpart. The failure to achieve a high GI in double planar implants is presumed to relate to technical difficulties rather than variation in individual performance. CONCLUSION: Our preliminary experience in treating early-staged tongue cancer with the HDR remote afterloading technique is inspiring, as it gives a local control rate of 100% with acceptable morbidity. Further studies are eagerly awaited to delineate the optimum schedule for this modality of treatment. It is hoped that the GI values, which represents the skills of insertion, could be routinely reported so that treatment results between different centers could be compared in a more precise manner. PMID- 9392553 TI - A technique for the use of afterloading 137Cs brachytherapy in renal-sparing irradiation of bilateral Wilms tumor. AB - PURPOSE: To describe a renal sparing brachytherapy technique for treating patients with bilateral Wilms tumor who have limited residual tumor measuring 2 cm or less after initial chemotherapy. METHODS AND MATERIALS: A technique for using brachytherapy in the radiotherapeutic management of bilateral Wilms tumor is described. Three patients with bilateral Wilms tumor were treated at our institutions. All three patients had initial nephrectomy of the contralateral kidney followed by chemotherapy. Local excision of residual tumor in the remaining kidney was done in all three cases. A 137Cs isotopic source was placed in the tumor bed at the time of the second surgery using a simple afterloading applicator. The techniques of applicator placement, localization, and brachytherapy dosimetry are described. The minimum tumor dose varied from 16 to 25 Gy. RESULTS: All three patients are alive and well at 28, 48, and 66 months after the procedure. There were no serious operative or postoperative sequelae. CONCLUSIONS: This simple brachytherapy technique was effective in selected cases of bilateral Wilms tumor where a renal-sparing radiotherapy approach was needed. This technique is most applicable when there is residual intrarenal tumor after partial nephrectomy, when the tumor is unifocal, and when the tumor bed is less than 2 cm diameter. PMID- 9392554 TI - Radiotherapy for cancer patients aged 80 and older: a study of effectiveness and side effects. AB - PURPOSE: To profile cancer patients aged 80 and older undergoing radiotherapy and to study the tumor response and side effects of therapy. METHODS AND MATERIALS: We retrospectively analyzed the records of patients aged 80 and older who received radiation therapy at James A. Haley Veterans Hospital and H. Lee Moffitt Cancer Center between 1988 and 1995. A total of 203 patients aged 80-94 received radiotherapy during this period. Treatment sites included head and neck [50], breast [16], chest [37], pelvis [53], and miscellaneous [39]. Age, treatment site, field size, total dose, response to treatment, treatment interruptions, incidence and severity of weight loss, myelosuppression, diarrhea, mucositis, dermatitis, and follow-up status are assessed using our departmental records and hospital tumor registry. RESULTS: Of 191 patients evaluated, 179 (94%) completed the treatment without serious complications. A total of 195 sites were irradiated. Twelve patients (6%) required interruption of the treatment. Therapeutic responses were seen in 86 out of 112 patients (77%) treated with curative intent (with 67% complete response) and in 67 out of 83 patients (81%) treated with palliative intent. The causes of treatment interruptions included weight loss from diarrhea, dysphagia, and progressive disease. Treatment interruptions were more likely in patients treated with large treatment fields. In patients treated for upper aero-digestive tract cancer, Grade 3 and 4 mucositis was noted in 20 and 2% of patients, respectively. Grade 1 and 2 enteritis was noted in 43% of patients treated for pelvic malignancies. Grade 3 dermatitis was noted only in 2% of patients. CONCLUSION: Radiotherapy is highly effective and well tolerated by the oldest old. Age is not a contraindication to aggressive radiotherapy. PMID- 9392555 TI - A prospective, randomized study addressing the need for physical simulation following virtual simulation. AB - PURPOSE: To accurately implement a treatment plan obtained by virtual or CT simulation, conventional or physical simulation is still widely used. To evaluate the need for physical simulation, we prospectively randomized patients to undergo physical simulation or no additional simulation after virtual simulation. METHODS AND MATERIALS: From July 1995 to September 1996, 75 patients underwent conformal four-field radiation therapy planning for prostate cancer with a commercial grade CT simulator. The patients were randomized to undergo either port filming immediately following physical simulation or port filming alone. The precision of implementing the devised plan was evaluated by comparing simulator radiographs and/or port films against the digitally reconstructed radiographs (DRRs) for x, y, and z displacements of the isocenter. Changes in beam aperture were also prospectively evaluated. RESULTS: Thirty-seven patients were randomized to undergo physical simulation and first day port filming, and 38 had first day treatment verification films only without a physical simulation. Seventy-eight simulator radiographs and 195 first day treatment port films were reviewed. There was no statistically significant reduction in treatment setup error (>5 mm) if patients underwent physical simulation following virtual simulation. No patient required a resimulation, and there was no significant difference in changes of beam aperture. CONCLUSIONS: Following virtual simulation, physical simulation may not be necessary to accurately implement the conformal four-field technique. Because port filming appears to be sufficient to assure precise and reliable execution of a devised treatment plan, physical simulation may be eliminated from the process of CT based planning when virtual simulation is available. PMID- 9392556 TI - A correlation between residual DNA double-strand breaks and clonogenic measurements of radiosensitivity in fibroblasts from preradiotherapy cervix cancer patients. AB - PURPOSE: To study the relationship between residual DNA damage and clonogenic measurements of radiosensitivity in fibroblasts from pretreatment cervix cancer patients. METHODS AND MATERIALS: Early passage vaginal fibroblasts from nine preradiotherapy cervix cancer patients and two radiosensitive skin fibroblast cell strains were studied. Cell survival was measured by clonogenic assay following both high and low dose rate irradiation. Residual DNA damage was measured using pulsed-field gel electrophoresis (PFGE) after irradiating radiolabeled, plateau-phase cells at 37 degrees C and allowing 24 h for repair. DNA damage was expressed both in terms of the residual damage slope (fitted to data from 60 to 150 Gy) and the fraction of activity released (FAR) following 150 Gy. RESULTS: The surviving fraction at 2 Gy (SF2) values after high dose rate irradiation for the vaginal fibroblasts ranged from 0.15 to 0.32 (a 2.2-fold difference). When the two radiosensitive cell strains were included, residual damage, expressed as the residual damage slope, correlated with alpha (r = 0.82, p = 0.002), D bar (r = -0.91, p < 0.001) and SF2 (p = -0.79, p = 0.004), and when the vaginal fibroblasts alone were studied, the residual damage slope again correlated with clonogenic survival, although less strongly [alpha (r = 0.66, p = 0.053), D bar (r = -0.83, p = 0.006), and SF2 (r = -0.63, p = 0.07)]. Within the group of vaginal fibroblasts there was a 4.0-fold difference in residual DNA damage slope. When residual damage was expressed as FAR at 150 Gy, then for all cell strains the correlations were alpha: r = 0.78, p = 0.004, D bar: r = -0.86, p = 0.001, and SF2: r = -0.78, p = 0.004, and for the vaginal fibroblast strains alone the correlations were alpha: r = 0.60, p = 0.088, D bar: r = -0.75, p = 0.02, and SF2: r = 0.62, p = 0.077. CONCLUSION: This study confirms previous findings that residual DNA damage correlates with clonogenic survival in fibroblasts. In addition, it demonstrates a correlation for fibroblasts from pretreatment cervix cancer patients demonstrating a relatively small range of SF2 values. PMID- 9392557 TI - Radiation enhancement by biochemical modulation and 5-fluorouracil. AB - PURPOSE: To evaluate the effects of biochemical modulation by N-(phosphonacetyl) L-aspartate (PALA), 6-methylmercaptopurine riboside (MMPR), and 6 aminonicotinamide (6AN), (PALA + MMPR + 6AN is referred to as PMA) on tumor radiosensitivity, and evaluate the efficacy of the addition of 5-FU to the PMA + XRT regimen for enhancement of tumor response to radiation without exceeding normal tissue tolerance. METHODS AND MATERIALS: A first generation transplant of the CD8F1 spontaneous murine tumor was studied. 31P nuclear magnetic resonance spectroscopy was used to determine the interval between chemotherapy and radiation based on energy depletion. PMA was administered three times with fractionated XRT (15 Gy x 3 = 45 Gy) on days 1, 10, or 11, and 21. The addition of 5-fluorouracil (5-FU) at maximum tolerated doses was evaluated and intergroup comparisons were made for tumor growth delay, local control, and disproportionate normal tissue damage. RESULTS: The combination of 5-FU + XRT induced a tumor doubling time of 75.4 days (67.4-84.4) (p < 0.0001 compared to XRT), validating that in this tumor model, pretreatment with bolus i.p. 5-FU enhanced XRT. In comparison, mice treated with PMA + XRT had a tumor doubling time (TDT) > 123.2 days (109.4-138.7), (p < 0.0001 compared to 5-FU + XRT). The addition of 5-FU to PMA + XRT induced a doubling time of > 170.8 days (150.7-193.7) (p = 0.0002 compared to PMA + XRT). The doubling time for the PMA + XRT cohort and the PMA + 5-FU + XRT cohorts are underestimates since some of the tumor bearing mice continue to have a complete regression (CR). The CR rate (measured on day 250) for the PMA + 5-FU + XRT cohort was 31.7% compared to 0% for 5-FU + XRT and 10% for PMA + XRT (p < 0.05). Mortality and local effects induced by radiation in the PMA + XRT group were comparable to the toxicity for the PMA + 5-FU + XRT group indicating that the addition of 5-FU at 75 mg/kg to PMA + XRT was tolerated and induced both greater CR and tumor doubling times than XRT alone, 5-FU (150 mg/kg) + XRT, or PMA + XRT. CONCLUSIONS: PMA is superior to 5-FU as a radiosensitizer in the schedule studied. The combination of PMA + 5-FU further enhanced XRT without exceeding normal tissue tolerance. PMID- 9392558 TI - The effect of UCN-01 (7-hydroxystaurosporine), a potent inhibitor of protein kinase C, on fractionated radiotherapy or daily chemotherapy of a murine fibrosarcoma. AB - PURPOSE: To investigate the effect of UCN-01 (7-hydroxystaurosporine), a potent and selective protein kinase C inhibitor, on fractionated irradiation or daily chemotherapy; cis-diamminedichloroplatinum(II) (cis-DDP) or 5-fluorouracil (5-FU) in vivo. Radiosensitivity and chemosensitivity given in combination with UCN-01 were further studied in vitro to analyze these in vivo results. METHODS AND MATERIALS: For in vivo studies, single-cell suspension was prepared from fourth generation FSa-II tumors and transplanted subcutaneously into the leg of 8-10 week-old C3Hf/Sed mice. Treatments were initiated when tumors reached an average diameter of 4 mm. Tumor response was studied using tumor growth and growth delay time assays. UCN-01 was given continuously for 7 days using Alzet osmotic pump (4.0 microg/microl/h or approximately 3.2 mg/kg/day). A daily gamma-ray dose of 10 Gy each was given in air for 7 days. Cis-DDP (0.7 mg/kg/day) or 5-FU (20 mg/kg/day) was given by an i.p. injection for 7 days. For in vitro studies, an established FSa-II cell line was used and cell survival was studied by colony formation assay. RESULTS: UCN-01 acted synergistically with fractionated irradiation, though it was slightly radioprotective in vitro and had no effect on SLD repair. The surviving fraction of the FSa-II cells treated with both UCN-01 and cis-DDP in vitro was lower than the calculated additive effect; however, the sensitizing effect of UCN-01 was not found when combined with either of the chemotherapeutic agents in vivo. Possible causes of synergism of combined UCN-01 and fractionated radiation may be that a continuous UCN-01 treatment inhibited clonogen repopulation during the course of fractionated irradiation and accumulated cells in the G2-M phase where cells are most sensitive to irradiation. CONCLUSION: UCN-01 is a promising agent that may indirectly interact with fractionated irradiation in vivo but may not with chemotherapeutic agents. PMID- 9392559 TI - Dose-response relationship for late functional changes in the rat brain after radiosurgery evaluated by magnetic resonance imaging. AB - PURPOSE: Only few quantitative data are available on late effects in the healthy brain after radiosurgery. An animal model can contribute to systematically investigate such late effects. Therefore, a model applying radiosurgery at the rat brain was established. A long-term (19 months) follow up study with 66 animals after radiosurgery was carried out. METHODS AND MATERIALS: In 60 animals, an area in the frontal lobe of the brain was irradiated stereotactically with a 15 MV linac. Different doses of 20, 30, 40, 50, and 100 Gy with two field sizes (3.9 and 5.9 mm collimator) were selected, using the integrated logistic formula with input parameters from human brain. The induced alteration of the blood-brain barrier permeability was investigated by means of contrast enhanced magnetic resonance imaging. RESULTS: A first intracranial signal enhancement was observed in one animal 160 days after irradiation with 100 Gy. Beginning at 5 months all animals in the two 100 Gy groups homogeneously showed contrast enhancement, but none of the other groups. This remained until 13 months after irradiation. The volume of contrast enhancement as well as the increase of signal intensity were different between the two 100 Gy groups. After 19 months, the animals irradiated with lower doses also showed contrast enhancements, although not uniformly throughout one group. A maximum likelihood fit of the logistic formula P(D) = 1/[1 + (D50/D)k] to the incidence of late effects for the 5.9 mm collimator at 19 months after irradiation results in the parameters D50 = 37.4(-5.2,+6.1) Gy and k = 4.7 +/- 2.4. CONCLUSIONS: An animal model was established to study late normal brain tissue response. The observed late effects appeared very similar to the estimation of the integrated logistic formula for human brain. Based on these radiosurgery techniques, future experiments will focus on modifications in the irradiation modalities, i.e., irregular volumes, radiation quality or fractionation. PMID- 9392560 TI - Modelling the optimal radiotherapy regime for the control of T2 laryngeal carcinoma using parameters derived from several datasets. AB - PURPOSE: A number of previous studies have used direct maximum-likelihood methods to derive the values of radiobiological parameters of the linear-quadratic model for head and neck tumors from large clinical datasets. Time factors for accelerated repopulation were included, along with a lag period before the start of this repopulation. This study was performed to attempt to utilise these results from clinical datasets to compare treatment regimes in common clinical use in the UK, along with other schedules used historically in a number of clinical series in North America and elsewhere, and to determine if an optimal treatment regime could be derived based on these clinical data. METHODS: The biologically-based linear-quadratic model, applied to local tumor control and late morbidity, has been used to derive theoretical optimum (maximising tumor control whilst not exceeding tolerance for late reactions) radiotherapy schedules based on daily fractions. The specific case of T2 laryngeal carcinoma was considered as this is treated primarily by radiotherapy in many centers. Parameter values for local control were taken from previous analyses of several large single-center and national datasets. A time factor and a lag period were included in the modelling. Values for the alpha/beta ratio for late morbidity were used in the range 1-4 Gy, which is compatible with the limited range of values reported in the literature for particular complications following radiotherapy for head and neck cancer. Early reactions and their consequential late morbidity were not modelled in this study, but assumed to be within tolerance. RESULTS: For treatments using daily fractions there was a broad optimum treatment time of between 3-6 weeks. The theoretical optimum depended to some extent on the value of the alpha/beta ratio for late morbidity, but in many cases was at or just beyond the end of the purported lag period of 3-4 weeks, although small values of alpha/beta between 1-2 Gy favour longer treatment times. Similar results were obtained using a range of parameter values derived from four independent clinical datasets. CONCLUSION: The mathematical modelling of this broad range of once-daily treatments for most of which differences in local control and late morbidity are essentially undetectable (< 5%) has shown how this clinically-recognised phenomenon is interpreted in terms of the combination of dose-response slopes, fractionation sensitivities and time factors for both tumor control and normal tissue morbidity. Although the conclusions are inevitably tempered by a number of caveats concerning confounding factors in different centers; for example, the use of different treatment volumes, the present analysis provides a framework with which to explore the potential value of modifications to conventional treatment schedules, such as the use of multiple fractions per day. PMID- 9392561 TI - Optimized beam planning for linear accelerator-based stereotactic radiosurgery. AB - PURPOSE: Current treatment planning for linear accelerator-based stereotactic radiosurgery and radiotherapy is a lengthy and iterative procedure. The planner has to manually select the beam arcs and carefully consider many different selections to ensure target volume coverage while sparing dose to critical organs. In this article we report an optimization procedure that can automatically select the beam arcs based on geometric and dosimetric analysis of the treatment parameters. METHODS AND MATERIALS: The optimization problem is introduced by using a Beam's Eye View (BEV) map where a pattern of lines represents a beam arc combination for a treatment plan. The collection of all possible treatment plans is described by using the concept of phase space where each point corresponds to a particular configuration of the system under consideration, and in this case, a particular beam arc combination. A geometric reduction of the phase space is performed by excluding static beam ports that irradiate too much critical organs and too little target volume. The phase space is further reduced by excluding beam arc combinations that do not comply with treatment convenience considerations and established planning experiences. These reductions significantly reduces the number of beam arc combinations to be considered and thus dramatically simplifies the computational complexity. The method of simulated annealing is then used to the reduced phase space to select the set of beam arcs that provides the best surface dose distribution for the target volume. The optimization procedure is applied to a radiosurgery case to compare the optimized beam arcs with the previously manually planned beam arcs. The procedure is also applied to 10 randomly selected cases for a comparison in terms of tissue-volume ratio calculations. RESULTS: The system is a highly automated beam arc planning tool for stereotactic radiosurgery and stereotactic radiotherapy. Its interactive nature allows the planner to rapidly consider many treatment plans to search for the best option. For the case presented, it is shown that the optimized beams substantially reduce the dose to the postrema. The tissue-volume ratio calculations demonstrate that the optimization often produces clinically superior treatment plans than the manual beam planning method. CONCLUSIONS: Our method of phase space reduction proves to be very useful in approaching the complex problem of treatment planning optimization. Not only does it substantially reduce the number of beam arcs that need to be considered, but it also simplifies the evaluation of the beam arc options. Both of these greatly reduce the computational complexity of the optimization and make the procedure fast and efficient. Moreover, the reduction of phase space adds another layer of interaction between the user and the beam selection procedure, so that the optimization process is well controlled and thus very effective. PMID- 9392562 TI - Total body irradiation with an arc and a gravity-oriented compensator. AB - PURPOSE: To deliver uniform dose distributions for total-body irradiation (TBI) with an arc field and a gravity-oriented compensator. This technique allows the patient to be treated lying on the floor in a small treatment room. METHODS AND MATERIALS: Through the sweeping motion of the gantry, a continuous arc field can deliver a large field to a patient lying on the floor. The dose profile, however, would not be uniform if no compensator were used, due to the effects of inverse square variation of beam intensity with distance as well as the slanted depth in patient. To solve this problem, a gravity-oriented compensator made of cerrobend alloy was designed. This compensator has a cross-section of an inverted isosceles triangle, with the apex always pointing downward, due to gravity. By properly selecting the thickness of the compensator, the width of the base, and the distance between the pivots to the base, the difference in the path length through the compensator can be made just right to compensate the effects of inverse-square and slanted depth, thus producing a uniform dose profile. RESULTS: Arc fields with a gravity-oriented compensator were used for 6, 10, 15, and 18 MV photon beams. The arc field can cover a patient with a height up to 180 cm. The field width was chosen from 32 to 40 cm at the machine isocenter. The optimal thickness of the compensator was found to be 2.5 cm, and its base was 25 cm wide. The distance from the pivot points to the flat surface of the compensator proximal to the beam ranges from 13 to 14 cm for different beam energies. The dose uniformity at a depth of 10 cm is within +/-5% for all beam energies used in this study. CONCLUSIONS: Highly uniform dose profiles for TBI treatments can be delivered with an arc and a gravity-oriented compensator. The proposed technique is simple and versatile. A single compensator can be used for all energies, because the amount of compensation can be adjusted by changing the distance to the pivot and/or the field size. PMID- 9392563 TI - Re: Clifford Chao et al., IJROBP 36(5):1039-1043; 1996. PMID- 9392564 TI - Regarding Aref et al., IJROBP 37:269-273; 1997. PMID- 9392565 TI - Measuring the "viral load" in cerebrospinal fluid in human immunodeficiency virus infection: window into brain infection? PMID- 9392566 TI - Cerebrospinal fluid human immunodeficiency virus type 1 RNA levels are elevated in neurocognitively impaired individuals with acquired immunodeficiency syndrome. HIV Neurobehavioral Research Center Group. AB - To determine whether cerebrospinal fluid (CSF) viral burden measurements can assist in the evaluation of human immunodeficiency virus (HIV)-associated neurocognitive disorders, we quantified HIV type 1 (HIV-1) RNA in CSF. Because previous findings suggested that disease stage, lymphocytic pleocytosis, and HIV 1 RNA levels in plasma may influence CSF viral burden, these variables were examined as potential modifying factors. HIV-1 RNA levels were quantified by using a reverse transcriptase-polymerase chain reaction assay. Performance on a comprehensive neuropsychological (NP) battery was noted in 97 prospectively enrolled, HIV-infected subjects. Among subjects with acquired immunodeficiency syndrome (AIDS) (<200 CD4+ lymphocytes), NP impairment was associated with significantly higher CSF RNA levels (3.1 vs 1.8 log10 copies/ml; p = 0.02); most impaired subjects met criteria for HIV-associated dementia or minor cognitive motor disorder. In subjects without AIDS, CSF RNA and NP impairment were unrelated. Before AIDS, CSF RNA was strongly correlated to plasma RNA and to pleocytosis, but in AIDS, CSF and plasma RNA were independent. In conclusion, we found elevated CSF HIV-1 RNA levels in NP impaired subjects with AIDS. Before AIDS, systemic viral replication, possibly through CD4+ mononuclear cell trafficking, may govern virus levels in CSF, whereas in AIDS, CD4 cell depletion may unmask a correlation between increased productive central nervous system HIV infection and clinical neurocognitive disorders. PMID- 9392568 TI - Long-term stroke risk in children with sickle cell disease screened with transcranial Doppler. AB - Stroke is an important complication of sickle cell disease. Stroke prediction is clinically important because it offers the possibility of primary prevention. In 1992, transcranial Doppler (TCD) evidence of elevated intracranial internal carotid or middle cerebral artery velocity was demonstrated to be associated strongly with an increased risk of ischemic stroke. This study extends the original study and includes 125 more children, longer follow-up, and intracranial hemorrhage in the stroke-risk model. Elevated time averaged mean maximum blood flow velocity, especially when velocity is 200 cm/sec or greater by TCD, was associated strongly with stroke risk. The cases not predicted by TCD point to the need for more information on the optimal timing of TCD surveillance for stroke risk. PMID- 9392567 TI - Relationship between human immunodeficiency virus-associated dementia and viral load in cerebrospinal fluid and brain. AB - Cerebrospinal fluid (CSF) human immunodeficiency virus (HIV) RNA levels were measured with the Nucleic Acid Sequence-Based Amplification (NASBA) assay to determine the relationship with neurological status; 37 subjects with HIV dementia (HIV-D) were compared with 77 with HIV with minor neurological signs (HIV-MCMD) and 93 neurologically normal HIV-seropositive individuals (HIV-NML). The NASBA assay had a lower limit of detection of 100 copies per milliliter. Mean CSF log HIV RNA levels were significantly higher in those with dementia after adjusting for CD4 count and were correlated with dementia severity. Plasma levels did not distinguish comparably immunosuppressed subjects with or without dementia. CSF and plasma RNA levels were significantly intercorrelated for subjects with CD4 counts <200/mm3 and also correlated inversely with CSF beta2 microglobulin. CSF RNA levels were independent of CSF pleocytosis or antiretroviral exposure. Brain RNA levels were consistently higher than CSF but correlated with CSF values for dementia subjects. The NASBA assay can be used reliably to determine HIV RNA levels in CSF, brain, and plasma samples. CSF HIV RNA may be a surrogate marker for brain infection, based on the observed correlation with brain levels. The association between plasma HIV RNA and CSF levels of HIV and beta2-microglobulin suggests that both viral load and CNS immune activation are important determinants of neurological disease. PMID- 9392569 TI - Ontogeny of ipsilateral corticospinal projections: a developmental study with transcranial magnetic stimulation. AB - Transcranial magnetic stimulation (TMS) has been used to describe the maturation of the corticospinal tract in children. Ipsilateral corticospinal connections have been demonstrated with TMS in patients with congenital mirror movements, in patients after hemispherectomy, and in children with hemiplegic cerebral palsy. The goal of the study was to find out whether corticospinal ipsilateral projections in children can be demonstrated during the first decade of life as part of normal ontogeny. For this purpose, we examined 50 normal children (age range, 3-11 years) with focal TMS over the left and right hemispheres to target muscles in proximal and distal parts of the upper extremity (first dorsal interosseus, biceps brachii, and brachioradialis). To lower the stimulation threshold, we stimulated under voluntary preinnervation. In two-thirds of the children we elicited ipsilateral motor evoked potentials (MEPs). This occurred more often in proximal than in distal muscles. The latency of the ipsilateral MEPs was about 12 to 14 msec longer than the usual contralateral response. From the age of 10, and in adults, ipsilateral MEPs could not be detected. Also considering lesion data from adult patients, the most likely explanation for the disappearance of ipsilateral corticospinal connections after the age of 10 years is an increasing transcallosal inhibitory influence during development. The presence of ipsilateral corticospinal connections appears to be a normal state in ontogeny. PMID- 9392570 TI - Differential distribution of the normal and mutated forms of huntingtin in the human brain. AB - Huntington's disease is an inherited disorder caused by expansion of a CAG trinucleotide repeat in the IT15 gene, which leads to expansion of a polyglutamine tract within the protein called huntingtin. Despite the characterization of the IT15 gene and the mutation involved in the disease, the normal function of huntingtin and the effects of the mutation on its function and on its neuronal location remain unknown. To study whether mutated huntingtin has the same neuronal distribution and intracellular location as normal huntingtin, we analyzed immunohistochemically both forms of this protein in the brain of 5 controls and 5 patients with Huntington's disease. We show that the distribution of mutated huntingtin is, like that of the normal form, heterogeneous throughout the brain, but is not limited to vulnerable neurons in Huntington's disease, supporting the hypothesis that the presence of the mutated huntingtin in a neuron is not in itself sufficient to lead to neuronal death. Moreover, whereas normal huntingtin is detected in some neuronal perikarya, nerve fibers, and nerve endings, the mutated form is observed in some neuronal perikarya and proximal nerve processes but is not detectable in nerve endings. Our results suggest that the expression or processing of the mutated huntingtin in perikarya and nerve endings differs quantitatively or qualitatively from the expression of the normal form in the same neuronal compartments. PMID- 9392572 TI - Long-term effects of pancreatic transplantation on diabetic neuropathy. AB - Restoration of a long-lasting euglycemic state by a functioning pancreatic transplantation (PTx) is the most logical treatment for insulin-dependent diabetes mellitus and for amelioration of secondary complications, including neuropathy. We evaluated neurological function by clinical examination, nerve conduction studies, and autonomic function tests in 115 patients with a functioning PTx and in 92 control patients treated with insulin, at baseline and 1, 2, 3.5, 5, 7, and 10 years later. In control patients, neuropathy progressively worsened during follow-up. The clinical examination score and composite indices of abnormality of motor and sensory nerve conduction decreased significantly at all intervals tested. Autonomic function indices also decreased, but significantly only after 1 year. In patients who received a successful PTx the neuropathy improved. The motor and sensory nerve conduction indices increased significantly at all intervals after transplantation, whereas the clinical examination and autonomic tests improved only slightly. Patients who received either a PTx alone, a PTx after a kidney graft, or simultaneous pancreatic and kidney transplantations improved similarly over the follow-up. These results indicate that a functioning PTx halts the progression and improves the signs of diabetic polyneuropathy by restoration of a normoglycemic state. PMID- 9392571 TI - Alterations in opioid receptor binding in Parkinson's disease patients with levodopa-induced dyskinesias. AB - Levodopa-induced dyskinesias remain a major challenge in the therapeutic management of Parkinson's disease (PD). Their etiology is unknown although dysfunction of striatal opioid transmission has been implicated in experimental models of PD. To determine whether the opioid system is involved in human dyskinetic PD, we measured in vivo opioid receptor binding in PD patients with and without levodopa-induced dyskinesias, using positron emission tomography (PET) and the opioid receptor ligand [11C]diprenorphine. Striatal and thalamic/occipital uptake ratios were calculated using a region of interest (ROI) approach. In addition, we used statistical parametric mapping (SPM) and images reflecting the volume of distribution of [11C]diprenorphine to assess changes in cerebral receptor binding on a voxel-by-voxel basis. By using the ROI approach, we found significantly reduced striatal and thalamic opioid binding in dyskinetic, but not in nondyskinetic, PD patients. The SPM approach confirmed reduced availability in these areas and, in addition, showed decreased cingulate and increased prefrontal opioid receptor binding in the dyskinetic patients. Our findings confirm that altered opioid transmission is part of the pathophysiology of levodopa-induced dyskinesias in PD and support further investigation into the role of opioid agents in the management of these involuntary movements. PMID- 9392573 TI - Proton magnetic resonance spectroscopic imaging and magnetic resonance imaging volumetry in the lateralization of temporal lobe epilepsy: a series of 100 patients. AB - Surgery is a safe and effective treatment for drug-resistant temporal lobe epilepsy (TLE). However, bilateral electroencephalographic (EEG) abnormalities are frequently present, making presurgical lateralization difficult. New magnetic resonance (MR) techniques can help; proton magnetic resonance spectroscopic imaging (MRSI) can detect and quantify focal neuronal damage or dysfunction based on reduced signals from the neuronal marker N-acetylaspartate, and magnetic resonance imaging (MRI)-based measurements of amygdala-hippocampal volumes (MRIVol) can improve the detection of atrophy of these structures. We performed proton MRSI and MRIVol in 100 consecutive patients with medically intractable TLE to determine how well these techniques agreed with the lateralization by extensive EEG investigation. We found that the EEG, MRSI, and MRIVol findings were highly concordant. The MRSI was abnormal in 99 of 100 patients (bilateral in 54%). The MRIVol was abnormal in 86 of 98 patients (bilateral in 28%). We obtained lateralization in 83% of patients using MRIVol alone, in 86% using MRSI alone, and in 90% by combining MRSI and MRIVol (vs 93% lateralization by EEG). MRSI was abnormal in 12 patients with normal MRIVol. The combination of proton MRSI and MRIVol can lateralize TLE accurately and noninvasively in the great majority of patients. By reducing reliance on EEG, these imaging techniques could reduce prolonged presurgical evaluation and make seizure surgery available to more patients. PMID- 9392574 TI - Entacapone improves motor fluctuations in levodopa-treated Parkinson's disease patients. Parkinson Study Group. AB - Motor fluctuations associated with levodopa therapy are common problems encountered in the long-term treatment of Parkinson's disease (PD). Entacapone, a peripherally acting, reversible inhibitor of catechol-O-methyltransferase, slows the elimination of levodopa in humans by reducing the formation of 3-O methyldopa. We conducted a placebo-controlled, double-blind, parallel-group, multicenter trial of entacapone in PD patients with motor fluctuations. Two hundred five patients were randomized to receive either entacapone 200 mg or matching placebo with each dose of levodopa and were followed for 24 weeks. The primary measure of efficacy was the change in percentage of "on" time (relief of parkinsonism) while awake, as recorded by subjects at home in diaries completed at 30-minute intervals. At baseline, patients averaged approximately 10 hours of "on" time per day while awake (60.5% "on" time), and entacapone treatment increased the percent "on" time by 5.0 percentage points. The effect of entacapone was more prominent in patients with a smaller percent "on" time (<55%) at baseline, and increased as the day wore on. Entacapone is effective at increasing the duration of response to levodopa and at relieving parkinsonism in patients experiencing motor fluctuations and was well tolerated during the 24 weeks of treatment. PMID- 9392575 TI - Quantitative neuropathology and quantitative magnetic resonance imaging of the hippocampus in temporal lobe epilepsy. AB - The aims of this study were to examine the relationships of hippocampal T2 (HCT2) relaxation time and magnetic resonance (MR)-based hippocampal volume (HCV) to neuronal (ND) and glial cell densities (GD) of hippocampal neuronal cell layers, and to obtain a better clinicopathological definition of hippocampal sclerosis (HS) and end folium sclerosis (EFS). Fifty-three hippocampi with HS, 6 with EFS, and 6 control hippocampi were studied. Pathologically, the HS group had a significantly higher logarithm (log) GD/ND than the controls in all hippocampal subregions, and than the EFS group in all subregions except the granule cell layer of the dentate gyrus (GCDG). The EFS group had a significantly higher log GD/ND than the control group only in the GCDG. Clinical correlations suggested that EFS may be the consequence of temporal lobe seizures and not an epileptogenic entity. Hippocampal atrophy in HS was associated with neuronal cell depletion and concomitant gliosis in the cornu Ammonis (CA) 1, CA2, CA3, and hilus. An increased HCT2 was associated with damage in the CA1 and also the hilus and has a different neuropathological basis than HCV loss. MR-based HCV measurement and HCT2 mapping, therefore, give complementary information in the presurgical evaluation of temporal lobe epilepsy and longitudinal studies. PMID- 9392576 TI - Effects of apomorphine on globus pallidus neurons in parkinsonian patients. AB - Current hypotheses of basal ganglia dysfunction in Parkinson's disease (PD) propose that neuronal hypoactivity in the globus pallidus externus (GPe), and hyperactivity in the output nuclei and the external and internal portions of the globus pallidus internus (GPi,e and GPi,i, respectively), result in the cardinal symptoms of PD. To test this theory, the nonselective D1- and D2-dopamine receptor agonist apomorphine (30-100 microg/kg SC) was administered to 14 levodopa-responsive PD patients who were off medication ("off" state) while recording neurons in GP. For 15 neurons that were continuously monitored, apomorphine was found to increase the firing rate of 3 neurons in GPe, and decrease the rate of 12 in GPi. The mean firing rates of many different neurons were determined before (n = 285) and at various intervals after (n = 184) the injection of the drug. The mean rates before apomorphine were as follows: GPe, 45 Hz (SD 15, n = 85); GPi,e, 67 Hz (SD 14, n = 125); and GPi,i, 85 Hz (SD 19, n = 75). At 25 to 35 minutes after APO, the rate of GPe neurons had increased to 72 Hz (SD 18, n = 7), the rate of GPi,e neurons had decreased to 39 Hz (SD 15, n = 15), and in GPi,i the rate decreased to 34 Hz (SD 22, n = 18). Eighty minutes after apomorphine administration, the mean firing rates returned to preadministration values. This study supports current models of basal ganglia dysfunction in PD and suggests that the therapeutic effect of apomorphine results from a normalization of the imbalance of neuronal activity in the direct and indirect pathways. PMID- 9392577 TI - Dietary fat intake and the risk of incident dementia in the Rotterdam Study. AB - A high intake of saturated fat and cholesterol and a low intake of polyunsaturated fatty acids have been related to an increased risk of cardiovascular disease. Cardiovascular disease has been associated with dementia. We investigated the association between fat intake and incident dementia among participants, age 55 years or older, from the population-based prospective Rotterdam Study. Food intake of 5,386 nondemented participants was assessed at baseline with a semiquantitative food-frequency questionnaire. At baseline and after an average of 2.1 years of follow-up, we screened for dementia with a three step protocol that included a clinical examination. The risk of dementia at follow-up (RR [95% CI]) was assessed with logistic regression. After adjustment for age, sex, education, and energy intake, high intakes of the following nutrients were associated with an increased risk of dementia: total fat (RR = 2.4 [1.1-5.2]), saturated fat (RR = 1.9 [0.9-4.0]), and cholesterol (RR = 1.7 [0.9 3.2]). Dementia with a vascular component was most strongly related to total fat and saturated fat. Fish consumption, an important source of n-3 polyunsaturated fatty acids, was inversely related to incident dementia (RR = 0.4 [0.2-0.91), and in particular to Alzheimer's disease (RR = 0.3 [0.1-0.9]). This study suggests that a high saturated fat and cholesterol intake increases the risk of dementia, whereas fish consumption may decrease this risk. PMID- 9392578 TI - Inflammatory central nervous system demyelination: correlation of magnetic resonance imaging findings with lesion pathology. AB - Magnetic resonance imaging (MRI) is widely used to evaluate and monitor disease activity in inflammatory demyelinating central nervous system (CNS) diseases such as multiple sclerosis. The present study aimed at correlating MRI findings with histological parameters in 6 cases of biopsy-proven inflammatory demyelination of the CNS. The earliest stages of demyelinating activity manifested as almost isointense lesions with a massive gadolinium-DTPA (Gd-DTPA) enhancement in T1 weighted scans. In T2-weighted scans, early active lesions formed a border of decreased intensity compared with the lesion center and the perifocal edema. The morphological correlate of this pattern in our patients was activated macrophages in the zone of myelin destruction at the plaque border. Late active lesions were hypointense in T1 and hyperintense in T2 scans. Inactive demyelinated and remyelinating lesions were hyperintense in T2 scans and enhanced inhomogenously after Gd-DTPA application. T1 scans revealed major differences in the degree of hypointensity that correlated with the extent of axonal damage, extracellular edema, and the degree of demyelination or remyelination. PMID- 9392580 TI - Clinical and magnetic resonance imaging findings in Batten disease: analysis of the major mutation (1.02-kb deletion). AB - A total of 36 patients with Batten disease (juvenile-onset neuronal ceroid lipofuscinosis), homozygous or heterozygous for the major mutation, a 1.02-kb deletion, in the CLN3 gene, were studied to relate their genotype to their clinical phenotype. The onset of visual failure and epilepsy was highly concordant in both groups. Great inter- and intrafamilial heterogeneity was demonstrated in the development of mental and physical handicap and in magnetic resonance imaging findings among both homozygous and heterozygous patients. The 1.02-kb deletion in homozygous form was always associated with mental and physical handicap, whereas the heterozygous phenotype could be extremely benign without affecting the intellectual level of the patient. Our data suggest that genetic background, modifying genes, and environmental factors all influence the final phenotype of Batten disease. PMID- 9392579 TI - Localization of frontotemporal dementia with parkinsonism in an Australian kindred to chromosome 17q21-22. AB - An Australian family with autosomal dominant presenile nonspecific dementia was recently described. The disease results in behavioral changes, usually disinhibition, followed by the onset of dementia accompanied occasionally by parkinsonism. Twenty-eight affected individuals were identified with an age of onset of 39 to 66 years (mean, 53 +/- 8.9 years). We mapped the disease locus to an approximately 26-cM region of chromosome 17q21-22 with a maximum two-point LOD score of 2.87. Affected individuals share a common haplotype between markers D17S783 and D17S808. This region of chromosome 17 contains the loci for several neurodegenerative diseases that lack distinctive pathological features, suggesting that these dementias, collectively referred to as frontotemporal dementia with parkinsonism linked to chromosome 17 (FTDP-17), are caused by mutations in the same gene. The entire coding region of five genes, mapped to the FTDP-17 candidate region, were also sequenced. This analysis included the microtubule-associated protein tau that is the major component of the paired helical filaments observed in Alzheimer's disease. No pathogenic mutations were identified in either the tau gene or in any of the other genes analyzed. PMID- 9392581 TI - Copper/zinc superoxide dismutase 1 and sporadic amyotrophic lateral sclerosis: analysis of 155 cases and identification of a novel insertion mutation. AB - Amyotrophic lateral sclerosis (ALS) is a progressive paralytic disorder resulting from the degeneration of motor neurons in the brain and spinal cord and leading to death within 5 years of symptom onset. The great majority of ALS cases are sporadic, with the familial form (FALS) representing fewer than 10% of all cases. Mutations in the copper/zinc superoxide dismutase 1 (SOD-1) gene have previously been identified as the underlying cause of approximately 20% of FALS cases. As the familial and sporadic forms of the disease are clinically similar, we have sought to determine whether such mutations in SOD-1 underlie any sporadic ALS cases. We have screened 155 sporadic cases by single-strand conformation polymorphism and have identified 4 sporadic cases that possess point mutations in exon 4 of the SOD-1 gene. Two of these mutations are identical to those previously reported in FALS cases. One mutation is novel, resulting in a frameshift at Val118 due to the replacement of G (first base in the last codon of exon 4) by AAAAC. This mutation results in a truncated SOD-1 protein due to the introduction of a stop codon three residues into exon 5. PMID- 9392583 TI - A novel muscle sodium channel mutation causes painful congenital myotonia. AB - Mutations in the skeletal muscle voltage-gated sodium channel alpha-subunit gene (SCN4A) have been associated with a spectrum of inherited nondystrophic myotonias and periodic paralyses. Most disease-associated SCN4A alleles occur in portions of the gene that encode the third and fourth repeat domains with the conspicuous absence of mutations in domain 1. Here we describe a family segregating an unusual autosomal dominant congenital myotonia associated with debilitating pain especially severe in the intercostal muscles. A novel SCN4A mutation causing the replacement of Val445 in the sixth transmembrane segment of domain 1 with methionine was discovered in all affected individuals and is the likely genetic basis for the syndrome. Myotonia was resistant to treatment; however, the most severely affected family member responded dramatically to the sodium channel blocking agent flecainide. PMID- 9392582 TI - Pallidotomy for hemiballismus: efficacy and characteristics of neuronal activity. AB - A patient with unremitting, medically intractable hemiballismus underwent a pallidotomy that abolished his involuntary movements. Firing rates of cells in the internal segment of the globus pallidus (GPi) recorded during this procedure were significantly lower than those observed during pallidotomy for Parkinson's disease, either "on" or "off" medication. Firing patterns in hemiballismus were characterized by low-frequency modulation of the firing rate. These results are consistent with the hyperkinetic model, which suggests that hemiballismus results from decreased inhibition of the pallidal relay nucleus of the thalamus by the GPi. The efficacy of surgery in the case of hemiballismus demonstrates that pallidotomy can be an effective treatment for this condition and suggests that patterned neuronal activity in the GPi is important in the mechanism of hyperkinetic disorders. PMID- 9392584 TI - SMI-31 immunoreactivity in inclusion body myositis. PMID- 9392585 TI - Clinically definite multiple sclerosis following optic neuritis. PMID- 9392586 TI - Involvement of cytokines in the histopathology of cerebral malaria. AB - Histopathologic and immunohistologic studies were performed in two cases of fatal cerebral malaria. On admission, both patients were in unarousable coma with hyperparasitemia. Examination of the tissue sections from various organs showed parasite sequestration in both cases with more extensive area of sequestration in case 1 than in case 2. A panel of monoclonal antibodies against cytokines applied to these tissues clearly detected tumor necrosis factor-alpha (TNF alpha), interferon-gamma (IFN gamma), interleukin-1beta (IL-1beta), and IL-10 in the tissues from brain and liver of case 1. A different cytokine profile, IL-4 and IL 10, was found in the brain tissues of case 2; no TNF alpha nor IFN gamma was detected. There was no cytokine detected in the tissues of other organs in either case. Results of the study suggest that histopathology in the brain of fatal cerebral malaria may be associated with focal accumulation of cytokines. Additionally, the type of cytokines produced locally in a particular tissue during malaria infection may be regulated by the degree of regional parasite sequestration. PMID- 9392587 TI - Prognostic significance of reduced red blood cell deformability in severe falciparum malaria. AB - Severe falciparum malaria is associated with microvascular obstruction resulting from sequestration of erythrocytes containing mature stages of the parasite. Since reduced red blood cell deformability (RBC-D) can contribute to impaired microcirculatory flow, RBC-D was measured in 23 patients with severe falciparum malaria (seven of whom subsequently died), 30 patients with uncomplicated malaria, and 17 healthy controls. The RBC-D, measured by ektacytometry, was significantly reduced in severe malaria and was particularly low in all fatal cases. At a low shear stress of 1.7 Pascal (Pa), a red blood cell elongation index less than 0.21 on admission to the hospital predicted fatal outcome with a sensitivity of 100% (confidence interval [CI] = 59-100%) and a specificity of 88% (CI = 61-98%). The reduction in the RBC-D appeared to result mainly from changes in unparasitized erythrocytes. Reduced deformability of unparasitized red blood cells in severe malaria may contribute to impaired microcirculatory flow and a fatal outcome in severe falciparum malaria. PMID- 9392588 TI - Molecular investigation of a multisource outbreak of Crimean-Congo hemorrhagic fever in the United Arab Emirates. AB - During the investigation of an outbreak of Crimean-Congo hemorrhagic fever (CCHF) in the United Arab Emirates (UAE) between 1994 and 1995, blood samples from suspected CCHF cases and ticks collected from livestock were tested for CCHF virus by antigen-capture ELISA and by a reverse transcription-polymerase chain reaction. Phylogenetic analysis of partial small (S) segment nucleotide sequences from four ticks and five human samples showed that with one exception, all the human and tick viruses clustered along with samples from Pakistan and Madagascar in one distinct lineage. Within this lineage, sequences from the UAE patients were identical or closely related to those from three Hyalomma spp. ticks obtained from livestock recently imported from Somalia. Another sequence from a UAE patient was more closely related to a CCHF virus from Nigeria. These data indicate that the 1994-1995 CCHF epidemic in the UAE was a multisource outbreak possibly associated with importation of CCHF virus-infected livestock and ticks. PMID- 9392589 TI - An outbreak of Crimean-Congo hemorrhagic fever in the United Arab Emirates, 1994 1995. AB - A multi-faceted investigation was conducted in the United Arab Emirates to characterize the epidemiologic and ecologic factors underlying an outbreak of Crimean-Congo hemorrhagic fever (CCHF) noted in November 1994 among abattoir workers. A chart review was conducted among hospitalized suspected cases of viral hemorrhagic fever with onset between January 1994 and March 1995 coupled with serologic testing of available specimens for the presence of virus antigen and IgG and IgM antibodies by ELISA. Livestock handlers and animal skin processors were interviewed and tested for the presence of IgG antibody. Sera from imported and domestic ruminants were examined for antibody for CCHF virus, and ticks collected from these animals were tested with an antigen-capture ELISA. Thirty five suspected cases of CCHF were identified (case fatality = 62%). Livestock market employees, abattoir workers, and animal skin processors accounted for 16 (57%) of 28 cases with known occupational status. Serologic evidence of past asymptomatic infection was noted in 12 (4%) of 291 livestock and abattoir workers but in none of the controls. Nineteen (7%) of 268 animals were positive for CCHF virus antibodies by ELISA including 12 ruminants from Somalia and Iran and five indigenous camels. One Hyalomma impeltatum and two H. excavatum from Somali cattle and one H. anatolicum from a Somali goat were positive for CCHF virus antigen. PMID- 9392590 TI - Prevalence of antibodies to western equine encephalomyelitis and St. Louis encephalitis viruses in residents of California exposed to sporadic and consistent enzootic transmission. AB - Sera from outpatients attending county health department clinics in areas of California with consistent (Imperial Valley) and sporadic (Sacramento Valley) enzootic transmission of western equine encephalomyelitis (WEE) and St. Louis encephalitis (SLE) viruses exhibited neutralizing antibody prevalence rates of 1.3% (n = 690) and 0.5% (n = 1,066) for WEE and 11.0% and 0.8% for SLE, respectively. Seroprevalence for SLE virus in Imperial County increased as a function of both age and years of residence, indicating that this virus was endemic with a low rate of annual infection. Of 26 sera that tested positive for SLE virus antibody by an enzyme immunoassay, but were negative by plaque reduction neutralization test, 14 (53%) had neutralizing antibody that reacted with > or = one type of dengue (DEN) virus. The DEN virus infections presumably were acquired elsewhere because neither the vectors nor DEN virus transmission occurs in California. The low prevalence of neutralizing antibody for WEE and SLE in the California human population indicated that despite recent increases in enzootic transmission, contact between humans and infectious mosquitoes have remained low. PMID- 9392591 TI - Prospective study of Entamoeba dispar infection in a cohort of mothers and their infants: relationship to serum antibody response. AB - The sera of cohorts of newborn infants and their mothers, characterized as cyst passers of Entamoeba with nonpathogenic zymodemes (E. dispar) and seropositive for amoebic antigens, were analyzed. Both cohorts were followed for a period of 12 months by microscopic examination of feces and determination of serum anti amoebic antibody titers using the indirect hemagglutination assay. Control groups (noncyst passer mothers and their infants) were included and followed. To characterize antigens involved in the induction of IgG and IgA antibody responses, Western blots of serum from all participants were tested and immunoplots of the frequency of antigenic recognition were constructed. Results of clinical follow-up and microscopic examination of feces showed that during the 12-month period none of the cyst passer mothers had episodes of diarrhea attributable to E. histolytica invasion; five of 21 children of cyst passer mothers became infected during the study, five of five infected children developed serum antiamebic antibodies (titers 1:64-1:128); none of the cohort of children from cyst passer mothers had diarrhea due to E. histolytica. Western blot analysis showed that there are antigenic fractions that induce serum antibodies of the IgG and IgA classes against E. dispar very early in the host parasite relationship. Our results suggest that mechanisms of antibody induction different from intestinal invasion may be operating in amebic infection. Intestinal absorption of antigen, systemic reflection of secretory antibody response, and priming of newborns by maternal anti-idiotypic antibody transfer are discussed. PMID- 9392592 TI - Genetic epidemiology of seropositivity for Trypanosoma cruzi infection in rural Goias, Brazil. AB - Chagas' disease is a zoonotic disease found throughout Latin America. Despite control programs in many of the affected countries, infection with Trypanosoma cruzi continues to be a major public health concern. In Brazil alone, approximately 53 million people live in endemic areas. Research with humans and with animal models indicates that there is variation in susceptibility to infection with T. cruzi. The reasons for this variation are not known although several studies have implicated genetic factors. An indirect immunofluorescence assay was used to assess seropositivity for T. cruzi infection in 716 adults from the municipality of Posse, Goias, Brazil. Detailed genealogic information was gathered at the time of sampling, which allowed assignment of 525 individuals to 146 pedigrees containing between two and 103 individuals; the remaining 191 unrelated individuals were retained as independents in the analysis. Using a maximum likelihood variance decomposition approach, we performed quantitative genetic analyses to determine if genetic factors could partially account for the observed pattern of seropositivity. The maximum likelihood estimate of the heritability of T. cruzi infection was 0.56 +/- 0.27 (mean +/- SE), indicating that genetic factors account for more than half of the observed variation in infection status. An additional 23% of the variation (c2 = 0.23 +/- 0.09) is attributable to the effects of shared environment, as assessed by common household. The results indicate that genetic factors play an important role in determining epidemiologic patterns of T. cruzi infection. Further characterization of these genetic factors may suggest new biologic areas to be targeted by prevention and intervention programs. PMID- 9392593 TI - Natural history of Giardia lamblia and Cryptosporidium infections in a cohort of Israeli Bedouin infants: a study of a population in transition. AB - The natural history of Giardia lamblia and Cryptosporidium infections were determined in a cohort of 164 Bedouin children, from a population not previously studied, which is in transition from nomadism to a settled life style. Stools were sampled monthly from birth to two years of age and at all diarrhea episodes. The risk of infection with G. lamblia and Cryptosporidium infection by age two was 91.5% and 48.8%, respectively. Cryptosporidium prevalence was 3-4% at all ages, whereas G. lamblia prevalence was > 30% after age one. Giardia lamblia and Cryptosporidium asymptomatic detection rates were high, 28.5% and 1.6%, respectively. Detection of G. lamblia was higher in diarrhea episode samples obtained before six months of age, but after that age and overall, the detection was lower than in nondiarrhea samples (odds ratio [OR] = 0.8, 95% confidence interval [CI] = 0.7-0.9, P < 0.05). Detection rates of C. parvum were higher in episode-related samples in all age groups (OR = 2.8, 95% CI = 1.9-4.2, P < 0.05) and infections in boys were more frequently symptomatic than in girls. While G. lamblia does not appear to be a consistent pathogen in this population where it is hyperendemic, Cryptosporidium has been shown to be an important cause of diarrhea in young children in the community. PMID- 9392594 TI - The Matola malaria project: a temporal and spatial study of malaria transmission and disease in a suburban area of Maputo, Mozambique. AB - A temporal and spatial study of malaria transmission in a suburban area of Maputo, Mozambique with a mean population density of 2,737/km2 was made from December 1992 to June 1995. A steep but continuous gradient was observed in the Plasmodium falciparum prevalence from 59.0% adjacent to the breeding sites to 5.4% only a few hundred meters distant. The entomologic inoculation rate ranged from a number too low to be determined in some districts to 20 infectious bites per person per year in the others. The risk of malaria was 6.2 times higher for individuals living less than 200 meters from the breeding sites than for individuals living 500 meters or more away from the breeding sites. In areas of high human density, mosquito and parasite dispersion is very limited, and therefore malaria control strategies could be more specifically targeted. PMID- 9392595 TI - Pediatric malaria in Houston, Texas. AB - We retrospectively reviewed the medical records of all infants and children (< 18 years of age) with the discharge diagnosis of malaria who were admitted to the four major pediatric teaching hospitals in Houston, Texas from January 1988 through December 1993. Thirty-four cases of pediatric malaria were identified in three newborns, 22 travelers, and nine recent immigrants. The travel destination was West Africa in 68%, Central America in 14%, India in 14%, and unknown in 4%. The location of the child's and parents' birthplace was available in 77% of the travel-related cases and in all cases the destination of travel was the parents' country of origin. The peak incident of the travel-related cases was late summer and early January corresponding to return from summer or Christmas vacation. Sixteen (75%) of the 22 travel-related cases had received either no prophylaxis (12 of 22) or inadequate (4 of 22) chemoprophylaxis. Half of the patients who were given appropriate chemoprophylaxis admitted to poor compliance. The clinical presentation was usually nonspecific. Fever was the most common symptom (97%) and was paroxysmal in one-third. Splenomegaly was the most common physical finding (68%). The malaria species identified included Plasmodium falciparum (56%), P. vivax (23%), P. malariae (3%), and unidentified (18%). Moderate anemia (hemoglobin level = 7.0-10 g/dL) occurred in 38% and severe anemia (hemoglobin level < 7.0 g/dL) in 29%. Three patients required transfusion. There were no end organ complications. In summary, pediatric malaria in Houston was primarily seen in immigrants or children of immigrants who returned to their native country. Education and preventive strategies should target these families and should be part of the routine well child care of these children. PMID- 9392596 TI - Human schistosomiasis in Puerto Rico: reduced prevalence rate and absence of Biomphalaria glabrata. AB - A combined epidemiologic and malacologic survey of schistosomiasis in Puerto Rico was carried out in areas where previous surveys had reported the prevalence of the disease. This limited survey, with 495 persons examined, found a low prevalence (0.6%) of Schistosoma mansoni infections. The infections were restricted to three people more than 36 years of age. No infections were detected in children 16 years of age or less, and this cohort comprised 57.8% of the study group. Malacologic surveys of the four streams, 10 rivers, and eight lakes throughout the island revealed the absence of intermediate host Biomphalaria glabrata and the presence of Thiara granifera, a competitive species of B. glabrata and the predatory snail Marisa cornuarietis. We believe that the absence of B. glabrata is the primary reason for the sustained reduction in the prevalence of schistosomiasis in Puerto Rico. PMID- 9392597 TI - Short report: surveillance of rickettsial infections in Indonesian military personnel during peace keeping operations in Cambodia. AB - Indonesian peacekeepers in Cambodia provided a unique study population to estimate the threat of rickettsial exposure to Rickettsia typhi (murine typhus), Orientia tsutsugamushi, (scrub typhus), and R. conorii (spotted fever) for the region. Prescreening prevalence measure showed a large proportion (36%) of soldiers with antibodies to R. typhi. Predeployment prevalence for antibodies to O. tsutsugamushi was 8%, with no evidence of background R. conorii infections. Actual seroconversions of R. typhi (3) and O. tsutsugamushi (1), attributed to exposure(s) in Cambodia, translated into annualized incidence rates of 24 and 8 per 1,000 per year, respectively. Surveillance of rickettsial infections and/or disease is particularly warranted in Cambodia with recent recognition of drug resistant scrub typhus in neighboring Thailand. PMID- 9392598 TI - Intraspecimen fecal egg count variation in Schistosoma mansoni infection. AB - To determine the degree of intraspecimen fecal egg count variation in Schistosoma mansoni infection and its impact on commonly used parasitologic parameters obtained by single egg counts, 10 25-mg Kato-Katz slides were prepared from each of three stool specimens collected on different days in a study group of 20 infected people. Individual fecal egg counts in these series of examinations varied considerably and this had profound consequences for the reliability of both qualitative and quantitative diagnosis. In light infections, S. mansoni eggs in stools appeared to be homogeneously mixed. However, this distribution became heterogeneous as the intensity of infection increased, indicating clustering of eggs in stool. The cumulative egg counts in the 10 slides of the same 20 people examined in this study were compared with those in 14 slides prepared from seven stool samples collected on different days. This revealed significantly different mean egg counts for six people, even after such exhaustive series of examinations. Intraspecimen variation also biased considerably some operational parameters used to determine the infection status at the group level, particularly when these were determined by the examination of single 25-mg slides. The examination of duplicate or multiple slides improved the intraspecimen estimates of these parameters but did not overcome day-to-day variation. The examination of fewer samples taken on different days proved to be more adequate than examining more slides from one stool specimen for the determination of precise estimates of the real infection status. PMID- 9392599 TI - Distribution, diversity, and host specificity of Bartonella in rodents from the Southeastern United States. AB - A number of Bartonella isolates were obtained from seven species of rodents sampled from 12 geographic sites representing the major biotic communities of the southeastern United States. Bartonella were isolated from the blood of 42.2% of 279 tested rodents. The highest prevalence of infection typically occurred among the most commonly captured species in the rodent community. Four phylogenetic groups, uniting 14 genotypic variants of Bartonella, were identified by sequence analysis of the citrate synthase gene. The level of sequence homology between genotypic groups varied from 88.8% to 96.4%, and the degree of homology among variants within groups was > or = 97%. Cotton rats (Sigmodon hispidus) harbored up to three phylogenetic groups of Bartonella at a single site, and Bartonella of two phylogenetic groups were isolated from a single rodent. All the Bartonella isolated from three species of Peromyscus clustered in a single distinct phylogenetic group, suggesting some host specificity may occur. Mouse ascitic fluids produced in BALB/c mice inoculated with Bartonella of three phylogenetic groups demonstrated high indirect fluorescent antibody (IFA) titers to homologous antigens. However, use of eight Bartonella antigens in an IFA test with sera from 394 wild-caught rodents resulted in either little or extremely low titers of antibody. PMID- 9392600 TI - Acquisition of Lyme disease spirochetes by cofeeding Ixodes scapularis ticks. AB - Ixodes scapularis ticks acquired spirochetes while cofeeding with Borrelia burgdorferi-infected nymphs (donors) on uninfected naive gerbils. Overall, 19% (67 of 345) of the recipient nymphs randomly exposed to gerbils with donor nymphs acquired spirochetes by fluorescent antibody (FA) tests, 18% (62 of 345) by the polymerase chain reaction. In a second experiment, donor nymphs were placed on the left ears, recipient nymphs were placed on the left and right ears, and xenodiagnostic larvae were placed on the gerbils' backs. Only recipient nymphs and larvae removed from the left ears were infected with B. burgdorferi. Infection rates were 47% (9 of 19), 87% (13 of 15) and 88% (14 of 16) in recipient nymphs placed on gerbils 0, 3, or 5 days after the donor nymphs, respectively, and 48% (12 of 25) in the larvae by FA analysis. Spirochetes appear to be acquired by cofeeding ticks from a localized infection near the feeding site rather than from a disseminated infection in the skin or blood. PMID- 9392601 TI - Differential effects of human serum and cells on the growth of Plasmodium falciparum adapted to serum-free in vitro culture conditions. AB - A single Plasmodium falciparum isolate was adapted for growth in serum-free culture medium. The parasitemia increased from 0.5% to 20% on day 7 after thawing. The asexual forms of the parasites appeared morphologically normal and pigment formation was comparable with that seen under standard conditions with serum present. Parasites were coincubated in 96-well plates with serum, peripheral blood mononuclear cells (PBMC), and PBMC in the presence of autologous serum from healthy non-immune individuals (n = 12), healthy semi-immune individuals (n = 12), and malaria patients (n = 7). Growth was monitored for six days. The concentration of interleukin-6 and interferon-gamma (IFN-gamma) in supernatants from the continuous cultures were measured by a bioassay and an enzyme-amplified sensitivity immunoassay. The results of this study showed that parasites cultured in serum-free medium in the presence of PBMC develop more rapidly, particularly with cells from malaria patients, compared with parasites cultured alone. The growth of parasites was different if 10% autologous serum was added to the culture. Parasite growth with sera from acutely infected individuals was similar with that with sera from aparasitemic, nonimmune individuals, and both supported significantly higher parasite growth over the six-day culture period compared with sera from the uninfected semi-immune individuals. Production of IFN-gamma by cells from nonimmune individuals and malaria patients was higher when cultures did not contain autologous serum. Nonimmune donor cells produced high amounts of IFN-gamma, but cells from the semi-immune donors produced little of this cytokine. There was no marked inhibition of parasite growth with any combination of serum and cells over six days of culture. A difference between the groups was observed after two days of culture, when growth with cells and serum from the uninfected, semi-immune group was significantly lower than that from the nonimmune group, but this was not subsequently sustained. The results of the study show that continuous cultivation of P. falciparum in serum-free medium provides a novel in vitro model to study mechanisms of the interplay between components of the human immune system and the malarial parasite, in which any possible influence of human serum is removed. PMID- 9392602 TI - Risk factors for cholera infection in the initial phase of an epidemic in Guinea Bissau: protection by lime juice. AB - Previous studies of cholera transmission have been conducted in the middle or at the end of an epidemic. Since modes of transmission could be different in different phases of an epidemic, we initiated a case-referent study immediately after the first cases had been hospitalized in a recent cholera epidemic in Guinea-Bissau in West Africa in October 1994. The cases investigated were consecutive adult patients resident in the capital of Bissau who were admitted the the National Hospital during the first two weeks of the epidemic. Referents were matched for district, gender, and age. The study showed a protective effect of using limes in the main meal (odds ratio [OR] = 0.2, 95% confidence interval [CI] = 0.1-0.3) and having soap in the house (OR = 0.3, 95% CI = 0.1-0.8). Not eating with the fingers and using water from a public standpipe were also protective. No specific source or mode of transmission was identified. Thus, cholera control programs in Africa may have to emphasize general hygienic conditions and the use of acidifiers in food preparation. PMID- 9392603 TI - Kinetic study of Russell's viper venom in envenomed patients. AB - Serum levels of Russell's viper venom in 30 patients bitten by Russell's viper were measured by an ELISA. In the initial serum samples, which were collected immediately after admission to the hospital (0.5-19 hr after the bite), venom was detected in 24 patients (80%), with levels ranging from 3 to 92 ng/ml. These levels correlated well with the patient's clinical signs. At 6-12 hr after antivenom therapy, the venom levels in most of the serum samples (21 of 24, 87%) had decreased to undetectable levels. In the remaining patients, whose serum venom levels could be detected 36-72 hr after therapy, the levels varied according to the effect of the antivenom therapy and the release of venom from a deposit at the bite site. PMID- 9392604 TI - Prospective study of tetanus-induced acute renal dysfunction: role of adrenergic overactivity. AB - To assess the mechanisms related to tetanus-induced acute renal failure (ARF), 30 patients with tetanus had their renal function prospectively studied and factors possibly related to renal changes were evaluated during four weeks of hospitalization. Fifty percent of these patients had a glomerular filtration rate (GFR) < or = 50 ml/min in the first or second week of hospitalization (Group I) and 50% had a GFR > 50 ml/min throughout the entire hospitalization period (Group II). Age, gender, tetanus incubation time and tetanus onset time, hospitalization time, use of nephrotoxic drugs, need for mechanical ventilation with intermittent positive pressure, and presence of systemic infection were similar in both groups. None of the patients presented with oliguria. Autonomic nervous system (ANS) overactivity, characterized by intense variations in systolic and diastolic blood pressure, by increased heart rate and elevated urinary metanephrine excretion, was higher in Group I compared with Group II. Plasma renin activity, serum creatinephosphokinase levels, and myoglobinuria were not significantly different between the two groups. These results strongly suggest that tetanus induced ARF has a high prevalence, is characterized by early onset, and is probably related to ANS overactivity. PMID- 9392605 TI - Discovery of encysted Paragonimus westermani eggs in the omentum of an asymptomatic elderly woman. AB - We report a case of asymptomatic chronic infiltrate of the omentum by eggs of Paragonimus westermani in an elderly woman who had immigrated to Taiwan from mainland China 46 years ago. The patient had a habit of eating raw freshwater crabs from the lakes of eastern China during her period of residence in that country. She stopped eating raw crabs after coming to Taiwan 20 years ago. During surgery for a peptic ulcer complicated by severe bleeding in 1995, her omentum was found to contain many small nodules approximately 2 x 2 x 1.5 cm in size. Biopsy of the nodules revealed eggs of P. westermani embedded in necrotic debris surrounded by capsules. A sputum examination result was negative and a chest radiograph was normal. The majority of the nodules in the omentum were removed during the surgery and praziquantel was given. At the present time, the patient remains asymptomatic. PMID- 9392606 TI - Cellular and humoral immune responses of hydatidosis patients to Echinococcus granulosus purified antigens. AB - The cellular and humoral immune responses of 41 hydatidosis patients, 12 healthy uninfected individuals, and seven patients with other parasitic diseases were determined using serologic and lymphoproliferative assays (LPAs), respectively. Echinococcal antigens were obtained by gel filtration of crude hydatid cyst fluid (HCF) on a Sephadex G-200 column. The fractions contained either a mixture of antigens A plus B or antigen B alone that was further enriched by boiling. All the hydatidosis patients responded positively by LPA to either crude or purified echinococcal antigens: 95% of them responded to either crude HCF, or a mixture of antigens A plus B and 83% to antigen B alone. The degree of the response to crude HCF (mean stimulation index [SI] = 75.3) was higher than that of purified antigens (SI = 39.1 for a mixture of antigens A plus B and SI = 36.9 for antigen B alone). No positive LPA response was obtained with the control groups. Serologic examinations showed that 78% of cases were positive by immunoblot, 73% by indirect hemagglutination, and 46% by immunoelectrophoresis. No correlation between the degree of cellular and humoral responsiveness to both crude and purified echinococcal antigens was observed. Nine of the 41 patients examined who were serologically negative also developed a high lymphoproliferative (LP) response to either crude or purified echinococcal antigens. The LP response remained positive over a long period after successful treatment. No relationship was observed between the results of treatment and the LP response. The present study indicates that the LPA could be used as an additional tool for the diagnosis of hydatid disease, particularly in seronegative cases, although it is unsuitable for effective monitoring treatment/surgery. PMID- 9392607 TI - Characterization of a recombinant Onchocerca volvulus antigen (Ov33) produced in yeast. AB - A yeast (Saccharomyces cerevisiae) expression system has been adapted to produce reagent quantities of a major Onchocerca antigen, Ov33. Using a pool of monoclonal antibodies produced against third-stage larvae, a cDNA library constructed from adult O. volvulus worms was screened. Twenty-seven cDNAs were isolated, two of which had sequence homology to Ov33, a putative aspartyl protease inhibitor, which is the immunodominant antigen of O. volvulus. These cDNAs were expressed at high levels intracellularly or through the secretory pathway of S. cerevisiae. Localization studies using antisera produced against purified recombinant protein demonstrated that Ov33 is a very abundant parasite protein present in the hypodermis, muscle, and uterus of female worms, as well as in embryonic microfilariae. The soluble recombinant protein secreted by yeast (C71) demonstrated inhibitory activity against the aspartyl protease pepsin. Antibodies to the recombinant protein-mediated leukocyte adherence to and killing of skin microfilariae. The sensitivity of a diagnostic test using recombinant Ov33 was evaluated using sera from 441 patients. The mean sensitivities for the two recombinant constructs, C27 and C71, were 82.2% and 85.4%, respectively. The combined sensitivity using both recombinant proteins was 94%. PMID- 9392608 TI - Pesticides and breast cancer: fact or fad? PMID- 9392609 TI - Survival after treatment of small-cell lung cancer: an endless uphill battle. PMID- 9392610 TI - Genetic tests for many rare diseases headed for orphan status. PMID- 9392611 TI - Chernobyl "liquidators" show increased risk of suicide, not cancer. PMID- 9392612 TI - Panel makes point about acupuncture. PMID- 9392614 TI - What causes thyroid cancer? PMID- 9392613 TI - Treatments sought for intractable forms of thyroid cancer. PMID- 9392615 TI - NCI selects first Director's Consumer Liaison Group. PMID- 9392616 TI - A National Cancer Institute Workshop on Hereditary Nonpolyposis Colorectal Cancer Syndrome: meeting highlights and Bethesda guidelines. PMID- 9392617 TI - Biology of cachexia. AB - About half of all cancer patients show a syndrome of cachexia, characterized by loss of adipose tissue and skeletal muscle mass. Such patients have a decreased survival time, compared with the survival time among patients without weight loss, and loss of total body protein leads to substantial impairment of respiratory muscle function. These changes cannot be fully explained by the accompanying anorexia, and nutritional supplementation alone is unable to reverse the wasting process. Despite a falling caloric intake, patients with cachexia frequently show an elevated resting energy expenditure as a result of increases in Cori cycle (i.e., catalytic conversion of lactic acid to glucose) activity, glucose and triglyceride-fatty acid cycling, and gluconeogenesis. A number of cytokines, including tumor necrosis factor-apha, interleukins 1 and 6, interferon gamma, and leukemia-inhibitory factor, have been proposed as mediators of the cachectic process. However, the results of a number of clinical and laboratory studies suggest that the action of the cytokines alone is unable to explain the complex mechanism of wasting in cancer cachexia. In addition, cachexia has been observed in some xenograft models even without a cytokine involvement, suggesting that other factors may be involved. These probably include catabolic factors, which act directly on skeletal muscle and adipose tissue and the presence of which has been associated with the clinical development of cachexia. A polyunsaturated fatty acid, eicosapentaenoic acid, attenuates the action of such catabolic factors and has been shown to stabilize the process of wasting and resting energy expenditure in patients with pancreatic cancer. Such a pharmacologic approach may provide new insights into the treatment of cachexia. PMID- 9392618 TI - Environmental estrogen stimulation of growth and estrogen receptor function in preneoplastic and cancerous human breast cell lines. AB - BACKGROUND: DDT and polychlorinated biphenyls (PCBs), which are widespread in the ecosystem, can mimic estrogen-mediated cell activities. Thus, they can potentially interfere with many physiologic processes. We compared the effects of organochlorines belonging to the DDT and PCB families, alone and in combination, for their ability to influence the estrogen receptor-mediated activities in preneoplastic breast epithelial cells and breast cancer cells. METHODS: Multiple assay systems requiring functional estrogen receptor were employed to test estrogen-like activity of organochlorine ligands. Two-sided statistical tests were used to compare the data. RESULTS: p,p'-DDT, the predominant form of DDT in the environment, is a more potent estrogen than o,p'-DDT (P<.001), although it is less effective than o,p'-DDT in inhibiting the binding of estradiol (natural estrogen) to estrogen receptor. Among the PCBs, Heptachlor is estrogenic (in transient reporter assays; P< or =.001), whereas Aroclor 1221 and Aroclor 1254, both individually and in combination, are only weakly estrogenic. CONCLUSION: p,p'-DDT is the most effective organochlorine in regulating estrogen receptor mediated cellular responses. In estrogen receptor-positive breast cancer cells, p,p'-DDT evokes responses by itself and enhances the responses in collaboration with estradiol or o,p'-DDT. PMID- 9392619 TI - Second primary cancers related to smoking and treatment of small-cell lung cancer. Lung Cancer Working Cadre. AB - BACKGROUND: An increased risk of second primary cancers has been reported in patients who survive small-cell carcinoma of the lung. The treatment's contribution to the development of second cancers is difficult to assess, in part because the number of long-term survivors seen at any one institution is small. We designed a multi-institution study to investigate the risk among survivors of developing second primary cancers other than small-cell lung carcinoma. METHODS: Demographic, smoking, and treatment information were obtained from the medical records of 611 patients who had been cancer free for more than 2 years after therapy for histologically proven small-cell lung cancer, and person-years of follow-up were cumulated. Population-based rates of cancer incidence and mortality were used to estimate the expected number of cancers or deaths. The actuarial risk of second cancers was estimated by the Kaplan-Meier method. RESULTS: Relative to the general population, the risk of all second cancers among these patients (mostly non-small-cell cancers of the lung) was increased 3.5 fold. Second lung cancer risk was increased 13-fold among those who received chest irradiation in comparison to a sevenfold increase among nonirradiated patients. It was higher in those who continued smoking, with evidence of an interaction between chest irradiation and continued smoking (relative risk = 21). Patients treated with various forms of combination chemotherapy had comparable increases in risk (9.4- to 13-fold, overall), except for a 19-fold risk increase among those treated with alkylating agents who continued smoking. IMPLICATIONS: Because of their substantially increased risk, survivors should stop smoking and may consider entering trials of secondary chemoprevention. PMID- 9392621 TI - Phase I study of human chorionic gonadotropin given subcutaneously to patients with acquired immunodeficiency syndrome-related mucocutaneous Kaposi's sarcoma. AB - BACKGROUND: In vitro and in vivo clinical studies have shown that certain preparations of human chorionic gonadotropin have antitumor activity against Kaposi's sarcoma, the most common tumor in patients infected with human immunodeficiency virus type 1 (HIV-1). METHODS: A phase I trial was conducted in 18 male patients with acquired immunodeficiency syndrome-related Kaposi's sarcoma. Successive cohorts of six patients each received human chorionic gonadotropin (A.P.L.; Wyeth-Ayerst, Radnor, PA) subcutaneously at doses of 5000 IU daily (level I), 10,000 IU three times a week (level II), or 10,000 IU daily (level III). Toxic effects, changes in reproductive hormone levels, HIV-1 RNA plasma levels, and response to therapy were evaluated. RESULTS: A.P.L. treatment was well tolerated at all dose levels, and no maximum-tolerated, dose-defined toxic effects were observed at the highest dose tested. The most common side effects were weight gain, increased libido, and increased energy. A persistent increase in testosterone level and a persistent decline in luteinizing hormone and follicle-stimulating hormone levels were seen over time. Major responses were observed in six patients. Partial remissions (> or =50% decrease in lesion numbers, volume, or surface area) were observed at dose level I and dose level II (two patients each); biopsy-confirmed complete remissions (resolution of all lesions) were observed at dose level III (two patients). All but one major response have persisted from 207 to more than 515 days. Nine patients had stable disease lasting 10 weeks or longer. CONCLUSIONS: A.P.L. given at daily doses ranging from 5000 to 10,000 IU has antitumor activity in patients with acquired immunodeficiency syndrome-related Kaposi's sarcoma. A.P.L. can be given for more than 1 year with minimal side effects. Larger efficacy studies are warranted. PMID- 9392620 TI - Phase I study of continuous-infusion L-S,R-buthionine sulfoximine with intravenous melphalan. AB - BACKGROUND: Increased intracellular glutathione has long been associated with tumor cell resistance to various cytotoxic agents. An inhibitor of glutathione biosynthesis, L-S,R-buthionine sulfoximine (BSO), has been shown to enhance the cytotoxicity of chemotherapeutic agents in vitro and in vivo. We performed a phase I study of BSO administered with the anticancer drug melphalan to determine the combination's safety/tolerability and to determine clinically whether BSO produced the desired biochemical end point of glutathione depletion (<10% of pretreatment value). METHODS: Twenty-one patients with advanced cancers received an initial 30-minute infusion of BSO totaling 3.0 g/m2 and immediately received a continuous infusion of BSO on one of the following schedules: 1) 0.75 g/m2 per hour for 24 hours (four patients); 2) the same dose rate for 48 hours (four patients); 3) the same dose rate for 72 hours (10 patients); or 4) 1.5 g/m2 per hour for 48 hours (three patients). During week 1, the patients received BSO alone; during weeks 2 or 3, they received BSO plus melphalan (15 mg/m2); thereafter, the patients received BSO plus melphalan every 4 weeks. Glutathione concentrations in peripheral blood lymphocytes were determined for all patients; in 10 patients on three of the administration schedules, these measurements were made in multiple sections from tumor biopsy specimens taken before, during, and after continuous-infusion BSO. RESULTS: Continuous-infusion BSO alone produced minimal toxic effects, although BSO plus melphalan produced occasional severe myelosuppression (grade 4) and frequent low-grade nausea/vomiting (grade 1-2). This treatment also produced consistent, profound glutathione depletion (<10% of pretreatment value). The degree of glutathione depletion in peripheral lymphocytes was considerably less than that observed in tumor sections. CONCLUSIONS: Continuous-infusion BSO is relatively nontoxic and results in depletion of tumor glutathione. PMID- 9392622 TI - Growth inhibition of human ovarian cancers by cytotoxic analogues of luteinizing hormone-releasing hormone. AB - BACKGROUND: Receptors for luteinizing hormone-releasing hormone (LH-RH) are found in nearly 80% of human ovarian cancers. The chemotherapeutic agent doxorubicin can be linked to [D-lysine6]LH-RH to form a cytotoxic analogue (AN-152) that may have greater specificity for tumor cells. This study was conducted to investigate the effects of AN-152 on the growth of LH-RH receptor-positive OV-1063 human epithelial ovarian cancers. METHODS: Nude mice bearing human ovarian tumors, OV 1063 or UCI-107 (LH-RH receptor negative), were injected intraperitoneally with saline (control) or with equimolar doses of AN-152 or doxorubicin; experiments involving mice with OV-1063 tumors also included groups that were administered [D lysine6]LH-RH either alone or in combination with doxorubicin. Tumor volume, weight, doubling time, and burden (i.e., tumor weight/body weight) as well as tumor apoptotic and mitotic indices were determined. The levels of receptors for LH-RH and epidermal growth factor (EGF) and their messenger RNAs were measured by use of radioreceptor and reverse transcription-polymerase chain reaction assays, respectively. RESULTS: The growth of OV-1063 ovarian tumors in nude mice, as based on reduction in tumor volume, was inhibited significantly (all P<.05, two sided) 4 weeks after treatment with AN-152, even at the lowest dose tested (413 nmol/20 g weight); the toxic effects of an equivalent dose of doxorubicin caused substantial mortality. High-affinity receptors for LH-RH and EGF were found on cell membranes of OV-1063 cancers; however, after in vivo treatment with AN-152, LH-RH receptor-binding sites were not detectable and EGF receptors were reduced in number. The growth of UCI-107 ovarian cancers was not inhibited by AN-152. CONCLUSIONS: In nude mice bearing LH-RH receptor positive OV-1063 epithelial ovarian cancers, systemic administration of AN-152 is less toxic and inhibits tumor growth better than equimolar doses of doxorubicin. PMID- 9392623 TI - Re: Recent trends in U.S. breast cancer incidence, survival, and mortality rates. PMID- 9392624 TI - Re: A science for the art of consensus. PMID- 9392625 TI - Loss of imprinting of the IGF2 gene in a Wilms' tumor in an adult. PMID- 9392626 TI - Shiga toxin mode of action in E. coli O157:H7 disease. AB - Shiga toxins (Stx) are virulence factors produced by selected bacteria pathogenic for humans. These multicomponent protein complexes are among the more potent toxins known. As inhibitors of eukaryotic protein synthesis, these toxins selectively inactivate ribosomes in an enzymatic manner. Specificity of cell targeting is determined by the high-affinity binding of Stx to its receptor, a glycosphingolipid (Gb3) located in the plasma membrane or some eukaryotic cells. Elaborated by food-borne E. coli O157:H7 bacteria, isotypes of Stx (Stx1 & Stx2) are required for the ensuing vascular changes in humans, including hemorrhagic colitis and renal hemolytic uremic syndrome. Experimental therapeutic intervention of Stx-associated disease includes the Stx receptor immobilized on biologically inert particles designed for oral presentation. PMID- 9392686 TI - Immunophenotyping of B lymphocytes by multiparametric flow cytometry in bone marrow aspirates and peripheral blood of healthy adults. AB - Establishing reference ranges by multiparametric immunophenotyping of mature B cells in bone marrow and peripheral blood of healthy adults is of interest because the detection of bone marrow infiltration, persistance of light chain restriction as well as discrimination between reactice and malignant lymphocytes are important applications of B-cell immunophenotyping. To determine the pattern of antigens as expressed by malignant mature B lymphocytes, bone marrow aspirates and peripheral blood of healthy adults in the present study were investigated for the presence and percentage frequency of those antigens as defined for immunophenotyping of B-cells by the REAL-Classification. For this purpose analysis of CD19 positive B lymphocytes by Live Gate analysis was performed. The established two-color as well as three-color stainings will serve as a basis for future investigations designed to test multiparametric analysis of B lymphocytes in bone marrow aspirates and peripheral blood. In conclusion, all investigated antibodies stained in varying percentage frequency on B-cell subtypes and statistical significant differences could be considered only for the CD19/CD10 staining in bone marrow aspirates. On the basis of this analysis, all the reported lineage antigen combinations are present both in malignant B lymphocytes as well as normal B cells in considerable percentage frequency. These findings are of important interest for follow-up investigations of patients with non Hodgkin s lymphomas by multiparametric immunophenotyping. PMID- 9392687 TI - Decreased function of monocytes and granulocytes during HIV-1 infection correlates with CD4 cell counts. AB - Monocytes and neutrophils are involved in the primary immune response against opportunistic infections that occur during the progression of human immunodeficiency virus (HIV) infection towards development of acquired immune deficiency syndrome (AIDS). Phagocytic cells operate through the generation of reactive oxygen species which may be toxic for fungi, bacteria and viruses. In the present study we evaluated the function of monocytes and granulocytes in whole blood samples of 16 healthy controls, 12 HIV infected subjects who had not undergone significant infections and of 17 individuals with AIDS. Using flow cytometric methods we were able to determine phagocytosis and respiratory burst under conditions that reflect the normal environment of these cells. Compared with results in samples from controls, granulocytes and monocytes from asymptomatic HIV infected patients exhibited a significantly increased capacity to phagocytose bacteria. The production of reactive oxygen intermediates was in the normal range. In comparison to asymptomatic HIV infected individuals, patients with AIDS showed a significant reduction of phagocytosis and respiratory burst which correlated with the number of CD4+ cells. In comparison to controls, patients infected with HIV, whether they were symptomatic or not, revealed a significantly diminished number of oxygen radical producing cells compared with the number of phagocytic cells. These results indicate that monocytes and granulocytes show reduced antimicrobial activity even in early stages of HIV infection. This defect is only partly due to the HIV infection itself as neutrophils are not target cells for HIV. PMID- 9392688 TI - Biotransformation and uric acid lowering effect of benzbromarone in patients with liver cirrhosis - evidence for active benzbromarone metabolites? AB - The disposition of benzbromarone and its uric acid lowering effect were investigated in 8 patients with compensated liver cirrhosis in order to obtain evidence whether dose requirements differ from subjects with normal liver function. Following a single oral dose of 100 mg benzbromarone, the plasma concentrations of the parent drug and the two hydroxylated main metabolites M1 and M2 as well as uric acid were determined up to at least 72 h. All patients were found to be rapid benzbromarone eliminators. In patients 2-8 the extent of systemic availability of benzbromarone, as estimated by the average AUC(0 infinite), was similar to previous observations in healthy individuals, whereas the values of both metabolites M1 and M2 tended to be lower in patients with liver cirrhosis. Cmax of benzbromarone and M1 also were lower in patients, M2 was equivalent to the data in subjects with normal liver function. tmax and the plasma elimination half-life t(1/2) varied within the same range as previously observed in healthy individuals. One patient exhibited much higher values in AUC(0-infinite); and Cmax of benzbromarone and both metabolites, and in addition of the elimination half-life of M1 and M2, whereas the plasma elimination of benzbromarone itself was not delayed. An effect of altered liver function cannot be excluded in this patient. Ten hours after benzbromarone administration the mean plasma uric acid in patients 2-8 was reduced by 31.5% and in patient 1 by 44.2% as compared to pretreatment values. Baseline levels were not regained until 72 h. These data are compatible with a prolonged uric acid lowering effect of an active benzbromarone metabolite. Altogether, the present observations do not suggest dose adjustment to be necessary in patients with compensated liver cirrhosis Child A and B. PMID- 9392690 TI - The influence of electromagnetic fields on human brain activity. AB - Possible effects of electromagnetic fields on human brain activity were studied. In a single-blind, cross-over-designed and placebo-controlled study 36 volunteers were exposed firstly to an electromagnetic field originating form a MediLine "MEGA-WAVE 150/1" therapy instrument and secondly to a field originating from a mobile, digital tetlephone as used for wireless telecommunication. All volunteers also underwent a control experiment with no field exposure. Application of the MEGA-WAVE instrument caused an increase in EEG power in the frequency bands Alpha2, Beta1 and Beta2 during and after field exposure. Operation of the mobile telephone caused an increase in the same frequency bands with a delay of approximately 15 minutes after exposure. PMID- 9392689 TI - High incidence of antibodies to 5-hydroxytryptamine, gangliosides and phospholipids in patients with chronic fatigue and fibromyalgia syndrome and their relatives: evidence for a clinical entity of both disorders. AB - The fibromyalgia syndrome (FMS) is one of the most frequent rheumatic disorders showing a wide spectrum of different symptoms. An association with the chronic fatigue syndrome (CFS) has been discussed. Recently, a defined autoantibody pattern consisting of antibodies to serotonin (5-hydroxytryptamine, 5-HT), gangliosides and phospholipids was found in about 70% of the patients with FMS. We were therefore interested in seeing whether patients with CFS express similar humoral immunoreactivity. Sera from 42 CFS patients were analysed by ELISA for these antibodies, and the results were compared with those previously observed in 100 FMS patients. 73% of the FMS and 62% of the CFS patients had antibodies to serotonin, and 71% or 43% to gangliosides, respectively. Antibodies to phospholipids could be detected in 54% of the FMS and 38% of the CFS patients. 49% of FMS and 17% of the CFS patients had all three antibodies in parallel, 70% and 55%, respectively had at least two of these antibody types. 21% of FMS and 29% of CFS patients were completely negative for these antibodies. Antibodies to 5-HT were closely related with FMS/CFS while antibodies to gangliosides and phospholipids could also be detected in other disorders. The observation that family members of CFS and FMS patients also had these antibodies represents an argument in favour of a genetic predisposition. These data support the concept that FMS and CFS may belong to the same clinical entity and may manifest themselves as 'psycho-neuro-endocrinological autoimmune diseases'. PMID- 9392691 TI - Erythrocyte indices as screening tests for the differentiation of microcytic anemias. AB - Algorithms for the differential diagnosis of anemias are commonly based on suspected incidences and simple laboratory parameters. Especially microcytic anemias are diagnosed using algorithms created in Mediterranean or Northern American regions. In a West German region we observe relatively high diagnostic uncertainty regarding common forms of anemias. As a hypothesis, this may be a result of inadequate algorithms not designed for regions with high incidences of anemias of chronic disease and low incidences of thalassemias. To further elucidate diagnostic problems we here report the frequencies of anemias in university hospital outpatients. Based on these data, the diagnostic values of different erythrocyte indices and of red cell distribution width in the differential diagnosis of anemias were calculated. 4525 patient files were reviewed retrospectively. 872 patients presented with anemia, 107 of which were hereditary forms and 765 of other forms. In hereditary anemias the frequency of thalassaemias (50 patients) and corpuscular hemolytic anemias (49 patients) was the same. Nearly half of the other anemias were covered by anemias of chronic disease and true iron deficiency anemias. Several indices intended to separate thalassemias from other microcytic anemias were tested for relevance. Sensitivity was between 0.75 and 0.85. Specificity was between 0.78 and 0.95. Red cell distribution width was not significantly different between thalassemias and iron deficiency. Only a red cell distribution width above 17.0 resulted in a specificity for iron deficiency of 0.91. Red cell distribution width is considered to be an unreliable screening test in a population with a low incidence of thalassemias. The high incidence of anemias of chronic disease in the region investigated should lead to more complex diagnostic procedures than offered by blood count values alone. PMID- 9392692 TI - Metabolism of apolipoprotein B in primary moderate hypercholesterolaemia: effects of acipimox and cholestyramine therapy. AB - The effects of combined therapy with acipimox (1250 mg/day) and cholestyramine (20 g/day) were examined in a group of 7 subjects with primary moderate hypercholesterolaemia (total cholesterol >=7 mmol/L). Radiolabeled VLDL subfraction turnovers were performed at baseline, during acipimox therapy and during combined therapy. Acipimox and combined therapies lowered plasma low density lipoprotein (LDL)-cholesterol by 20% (P<0.001) and 27% (P<0.001) respectively. The marked fall in LDL-cholesterol associated with acipimox therapy, was due to a reduced production rate of LDL (apolipoprotein) apoB. This is shown to be a result of reduced direct LDL apoB production, reduced IDL to LDL transfer consistent with inhibition of hepatic triglyceride lipase, and with a reduction in the overall throughput of VLDL1 apoB. With combined therapy both reduced production and increased catabolism of apoB containing LDL precursors and of LDL itself have to be invoked to explain the fall in plasma LDL-cholesterol. PMID- 9392693 TI - Talc granuloma of the uterus. AB - A 40-year-old woman presented with acute obstructive lung disease. She was treated with steroids and achieved a good response. However, she had two previous pelvic operations with histological findings of uterine serosal caseating granulomas presenting in the latter operation. Antituberculosis (TB) prophylaxis was suggested due to the suspicion of TB reactivation under steroid management. Investigation for TB infection or exposure was negative while revision of the histological slides disclosed talc granulomas. Although talc granuloma is reported in starch powder gloves which should not contain talc, it is still very uncommon. PMID- 9392695 TI - Alcohol consumption and mortality among middle-aged and elderly U.S. adults. AB - BACKGROUND: Alcohol consumption has both adverse and beneficial effects on survival. We examined the balance of these in a large prospective study of mortality among U.S. adults. METHODS: Of 490,000 men and women (mean age, 56 years; range, 30 to 104) who reported their alcohol and tobacco use in 1982, 46,000 died during nine years of follow-up. We compared cause-specific and rates of death from all causes across categories of base-line alcohol consumption, adjusting for other risk factors, and related drinking and smoking habits to the cumulative probability of dying between the ages of 35 and 69 years. RESULTS: Causes of death associated with drinking were cirrhosis and alcoholism; cancers of the mouth, esophagus, pharynx, larynx, and liver combined; breast cancer in women; and injuries and other external causes in men. The mortality from breast cancer was 30 percent higher among women reporting at least one drink daily than among nondrinkers (relative risk, 1.3; 95 percent confidence interval, 1.1 to 1.6). The rates of death from all cardiovascular diseases were 30 to 40 percent lower among men (relative risk, 0.7; 95 percent confidence interval, 0.7 to 0.8) and women (relative risk, 0.6; 95 percent confidence interval, 0.6 to 0.7) reporting at least one drink daily than among nondrinkers, with little relation to the level of consumption. The overall death rates were lowest among men and women reporting about one drink daily. Mortality from all causes increased with heavier drinking, particularly among adults under age 60 with lower risk of cardiovascular disease. Alcohol consumption was associated with a small reduction in the overall risk of death in middle age (ages 35 to 69), whereas smoking approximately doubled this risk. CONCLUSIONS: In this middle-aged and elderly population, moderate alcohol consumption slightly reduced overall mortality. The benefit depended in part on age and background cardiovascular risk and was far smaller than the large increase in risk produced by tobacco. PMID- 9392696 TI - Epidural analgesia compared with combined spinal-epidural analgesia during labor in nulliparous women. AB - BACKGROUND: Among nulliparous women, there appears to be an association between the use of epidural analgesia during labor and an increased risk of dystocia. We tested the hypothesis that combined spinal-epidural analgesia, which permits ambulation during labor, is associated with a lower incidence of dystocia than continuous lumbar epidural analgesia. METHODS: Between July 1995 and September 1996, we randomly assigned 761 nulliparous women in spontaneous labor at term who requested epidural analgesia to receive either continuous lumbar epidural analgesia or a combination of spinal and epidural analgesia. Among the women who received combined spinal-epidural analgesia, some were discouraged from walking and others were encouraged to walk. Maternal and neonatal outcomes, the incidence of dystocia necessitating cesarean section, and measures of patients' satisfaction were compared in the two groups. RESULTS: There were no significant differences in the overall rate of cesarean section, the incidence of dystocia, the frequency of maternal or fetal complications, the patients' or nursing staff's assessment of the adequacy of analgesia, or the degree of overall satisfaction between the two groups. Significantly more women receiving combined spinal-epidural analgesia had pruritus (P<0.001) and requested additional epidural bolus doses of local anesthetic (P=0.01). For all the women, dystocia necessitating cesarean section was significantly more likely when analgesia was administered with the fetal vertex at a negative station (odds ratio, 2.5; P<0.001) or at less than 4 cm of cervical dilatation (odds ratio, 2.2; P<0.001). CONCLUSIONS: As compared with continuous lumbar epidural analgesia, the combination of spinal and epidural analgesia is not associated with an overall decrease in the incidence of cesarean delivery. PMID- 9392697 TI - The association of atopy with a gain-of-function mutation in the alpha subunit of the interleukin-4 receptor. AB - BACKGROUND: Atopic diseases are very common, and atopy has a strong genetic predisposition. METHODS: Using single-strand conformation polymorphism analysis and DNA sequencing, we searched for mutations in the a subunit of the interleukin 4 receptor that would predispose persons to atopy. We examined the prevalence of the alleles among patients with allergic inflammatory disorders and among 50 prospectively recruited adults. Subjects with atopy were identified on the basis of an elevated serum IgE level (> or = 95 IU per milliliter) or a positive radioimmunosorbent test in response to standard inhalant allergens. The signaling function of mutant interleukin-4 receptor a was examined by flow cytometry, binding assays, and immunoblotting. RESULTS: A novel interleukin-4 receptor alpha allele was identified in which guanine was substituted for adenine at nucleotide 1902, causing a change from glutamine to arginine at position 576 (R576) in the cytoplasmic domain of the interleukin-4 receptor alpha protein. The R576 allele was common among patients with allergic inflammatory disorders (found in 3 of 3 patients with the hyper-IgE syndrome and 4 of 7 patients with severe atopic dermatitis) and among the 50 prospectively recruited adults (found in 13 of 20 subjects with atopy and 5 of 30 without atopy; P=0.001; relative risk of atopy among those with a mutant allele, 9.3). The R576 allele was associated with higher levels of expression of CD23 by interleukin-4 than the wild-type allele. This enhanced signaling was associated with a change in the binding specificity of the adjacent tyrosine residue at position 575 to signal-transducing molecules. CONCLUSIONS: The R576 allele of interleukin-4 receptor alpha is strongly associated with atopy. This mutation may predispose persons to allergic diseases by altering the signaling function of the receptor. PMID- 9392698 TI - Effect of ticlopidine on the long-term patency of saphenous-vein bypass grafts in the legs. Etude de la Ticlopidine apres Pontage Femoro-Poplite and the Association Universitaire de Recherche en Chirurgie. AB - BACKGROUND: Optimal therapy to prevent late occlusion of arterial bypass grafts in the legs has not been determined. We assessed the effect of ticlopidine, an inhibitor of platelet aggregation, on the long-term patency of saphenous-vein bypass grafts for the treatment of peripheral vascular disease. METHODS: A total of 243 patients with femoropopliteal or femorotibial saphenous-vein bypass grafts were randomly assigned to receive either ticlopidine (250 mg twice a day) or matching placebo for two years. The primary end point was graft patency at two years, as assessed by physical examination, measurement of the ankle brachial index, and duplex ultrasonography or arteriography. The incidence of death and major ischemic events was also analyzed in the two groups. RESULTS: After two years, 66.4 percent of the patients were alive with a patent graft in the ticlopidine group, as compared with 51.2 percent in the placebo group (95 percent confidence interval for the difference between the two groups, 2.9 to 27.4 percent; P=0.02). The two-year cumulative patency rate was 82 percent in the ticlopidine group and 63 percent in the placebo group (P=0.002). There was no significant difference between groups in overall mortality or major ischemic events. CONCLUSIONS: Ticlopidine significantly improved the long-term patency of saphenous-vein bypass grafts in the legs. Since the drug was well tolerated, its use can be recommended after peripheral-vein bypass surgery. PMID- 9392699 TI - Images in clinical medicine. Subdural hematoma. PMID- 9392700 TI - Hepatitis B virus infection. PMID- 9392701 TI - Noninvasive ventilation. PMID- 9392703 TI - Prepublication release of Journal articles. PMID- 9392704 TI - Hazards and benefits of alcohol. PMID- 9392705 TI - Analgesia for labor. PMID- 9392706 TI - Interleukin-4 and the genetics of atopy. PMID- 9392707 TI - The rule of double effect--a critique of its role in end-of-life decision making. PMID- 9392708 TI - Electronic fetal heart rate monitoring: a primer for the critical care nurse. AB - Fetal assessment is an essential component of nursing care for a pregnant woman who is critically ill. If the fetus has reached the gestational age for which electronic fetal monitoring (EFM) is possible, intermittent or continuous fetal assessment with EFM may be used. Most nurses who specialize in adult intensive care nursing do not have the education or clinical experience to interpret EFM data. Collaboration with perinatal care providers is necessary to insure that fetal assessments are timely and accurate and that nursing interventions based on the data are appropriate. When perinatal providers participate as members of the team caring for critically ill pregnant women, terminology and physiologic parameters may be used that are not standard or routinely used in the ICU setting. The physiologic and hemodynamic changes that occur during pregnancy add another dimension to the nursing care required. This article reviews common terms used to describe fetal status and appropriate nursing interventions used in caring for the critically ill pregnant woman. PMID- 9392709 TI - Aggressive management of HELLP syndrome and eclampsia. AB - The nursing care of women critically ill during pregnancy is an ever-changing specialty that presents unique clinical problems and issues. The woman and the fetus can be profoundly affected by the hypertensive disorders of pregnancy. The underlying pathology must be identified in relation to its effects on the pregnancy, the effects of the pregnancy on the disease, and the implications for the woman and the fetus. Severe preeclampsia, the HELLP syndrome (hemolysis, elevated liver enzymes, and low platelets), and eclampsia present such challenges and management problems. This article discusses the pathophysiology and management implications of severe preeclampsia, the HELLP syndrome, and eclampsia. PMID- 9392710 TI - Support of the breast-feeding mother in critical care. AB - Physiologic stabilization and maintenance of life of a critically ill, newly delivered woman is the immediate priority for the critical care team. Once stabilized however, each mother must be evaluated for lactation status. For the mother who has chosen to bottle-feed her infant, the nurse should initiate nursing care measures to suppress lactation. With the mother who has chosen to breast-feed her infant, the nurse has additional responsibility. Because actual breast-feeding will most likely be suspended temporarily, the nursing staff should be knowledgeable in breast care associated with establishing a milk supply, expressing milk to prevent breast engorgement, and initiating actual breast-feeding when the mother's condition permits. This article provides the advanced practice nurse with information, skills, and resources necessary to assess, initiate, and maintain breast-feeding, one of the most important physiologic and psychologic needs of the mother-infant dyad. PMID- 9392711 TI - Meeting the challenges of critical care obstetrics in today's health care system: collaborative education and practice strategies. AB - The transformation of health care in the United States necessitates developing creative strategies to provide quality and cost-effective care for the critically ill obstetric patient population. A discussion of the care of these patients is presented in a multidisciplinary framework, using a case study to illustrate the management process. The significance of contributions and collaboration of advanced practice nurses in perinatology and critical care in the care management of the critically ill obstetric patient is described. Such strategies as location of the patient and educational preparation for nurses caring for these complex patients are offered for consideration. Implications are described and recommendations are made for administrative and clinical practice. PMID- 9392712 TI - A collaborative approach to fetal assessment in the adult intensive care unit. AB - When a pregnant woman is admitted to the adult intensive care unit (ICU), responsibility for fetal assessment must be assumed by a nurse who is competent in interpreting data obtained by auscultation of the fetal heart rate or by the electronic fetal monitor. The fetus is a distinct patient requiring assessments, interventions, and evaluation, including documentation of nursing care provided, similar to any patient in the ICU setting. Most ICU nurses do not have adequate knowledge and clinical experience to assume this responsibility. Therefore, in institutions in which critically ill pregnant women are transferred to the adult ICU, a formal plan should be in place that includes care provided by nurses who are competent in fetal assessment. This article describes a collaborative approach to ensure that fetal assessments are performed by nurses who have the experience and education to do so and includes common terminology used to describe fetal status so that ICU nurses are familiar with the language and appropriate nursing intervention. PMID- 9392713 TI - Maternal-fetal assessment of the critically ill parturient: decisions related to delivery. AB - Preparation for delivery of the critically ill pregnant women begins soon after admission to the intensive care unit. Unless maternal or fetal condition deteriorates, remaining in utero may be more beneficial to the premature fetus. A decision regarding the timing of delivery is based on the impact on maternal and fetal well-being of maintaining the fetus in utero. Maternal or fetal instability may necessitate immediate delivery, and specialist from critical care, obstetrics, neonatology, and anesthesiology should decide on the most appropriate location for labor and delivery. Personnel from affected departments are alerted as soon as possible to facilitate the gathering of necessary equipment and supplies and the attendance of skilled professionals in intrapartum management and neonatal resuscitation. PMID- 9392714 TI - Cardiopulmonary resuscitation: pregnant women are different. AB - Although cardiopulmonary arrest rarely occurs in the pregnant woman, it is important that the health care team know the appropriate actions to take in such an event, to promote positive outcomes for both mother and fetus. Specific techniques, personnel, and equipment are required to manage this grave situation. The principles of airway, breathing, and circulation are used as with any client in cardiopulmonary arrest; however, modifications must made because of the physiologic changes that normally occur during pregnancy. If the pregnant woman does not respond to treatment, a cesarean delivery must be attempted within 5 minutes of the arrest if uterine size indicates gestational age of at least 20 weeks. This article describes the adaptations of traditional cardiopulmonary arrest procedures required to treat the pregnant woman who sustains a cardiopulmonary arrest, protocols for managing the communication of the emergency code, emergency equipment that must be available, and the importance of teams in managing mother and neonate. PMID- 9392715 TI - Beneficence toward whom? Ethical decision-making in a maternal-fetal conflict. AB - Ethical dilemmas and conflicts occur frequently in critical care units. When these dilemmas involve a pregnant patient, the conflicts are further complicated, because they also involve the interests of the fetus. Using an ethical decision making process facilitates the analysis of ethical dilemmas and their resolutions. This process is used to analyze the dilemma of selecting appropriate treatment for a woman at 30 weeks' gestation, diagnosed with acute lymphocytic leukemia. The case is examined from the perspective of the mother and the fetus, using the decision-making process. The medical indications include the patient's physical state, disease process, and treatment options. Patient preferences are the ethical and legal center of the patient-physician, and patient-nurse relationship. Contextual features include religious beliefs, cultural values, family dynamics, and financial and legal aspects of the care options. Finally, the ethical principles, relevant and in conflict, are assessed. Exploring these areas clarifies the best treatment option in consideration of the issues and facts of the case. PMID- 9392716 TI - Neutralizing ageism in critical care via outcomes research. AB - Ageism, as a mind-set, amplifies a belief that intensive care for the elderly is ineffectual. However, there are little data to support the notion that advanced chronological age alone predicts unfavorable outcomes in response to intensive care. A lack of outcome data, combined with ageism, may place older patients at risk for rationing of intensive care. Currently, neither public policy nor cultural norms directly support a limitation in services to the elderly. However, as pressure to reduce health care costs increases, critically ill elderly patients may be targeted for rationing. In this context, outcomes research involving elderly populations is crucial. The purpose of this report is to explicate the risk of ageism in the delivery of intensive care and to describe methods for implementing outcomes assessment for critically ill elderly patients as an essential element in a continuum of care. PMID- 9392717 TI - Impact of a gerontological nurse practitioner on the nursing home elderly in the acute care setting. AB - A retrospective analysis of the length of stay of nursing home elderly patients admitted to the hospital demonstrates a significant decrease when patients are comanaged by a nurse practitioner and attending physicians. A comparison of the 20 most heavily populated diagnostic groups between 1993 and 1994 reveals a shorter average length of stay in 17 of the 20 diagnostic groups, and an overall mean decrease of 2.78 days for all groups. This article discusses the impact of the gerontological nurse practitioner on hospital length of stay of the nursing home elderly with acute illness. Research is needed to document advanced practices that decrease length of stay, as well as to control for extraneous variables. PMID- 9392718 TI - Current trends in the clinical management of an old enemy: congestive heart failure in the elderly. AB - Much has been learned in the past decade, through quality medical and nursing research, regarding the pathophysiology and management of congestive heart failure. Primary care practitioners are challenged to apply the latest data to the clinical management of these patients when there is clear evidence of improvement in the control of symptoms and quality of life. Because of the complexities of the disease process, older adults with congestive heart failure require a comprehensive approach that is particularly well suited to the knowledge and skills of the advanced practice nurse. PMID- 9392719 TI - Decreasing polypharmacy in clients most at risk. AB - Approximately one third of all drugs prescribed in the United States are considered unnecessary. Polypharmacy, the unnecessary, excessive use of prescription and over-the-counter medications, increases clients' risk for adverse drug reactions and drug-drug interactions. Reducing the incidence of polypharmacy is a health protection goal of Healthy People 2000. Older clients, particularly older women living independently in the community, are at highest risk for polypharmacy caused by age-related changes in pharmacokinetics and pharmacodynamics, the presence of comorbidities requiring pharmacologic management, and high rates of unintentional noncompliance with their therapeutic regimen. Health providers may contribute to polypharmacy directly by excessive or inappropriate prescribing practices or indirectly through their inability to resist client's demands for pharmacologic interventions. Strategies to prevent and detect polypharmacy are suggested to reduce its incidence and the severity of its consequences. PMID- 9392720 TI - Case management of critically ill elders: a case study. AB - As the number of elderly increase, new challenges for the management of their health care arise. Elders who become critically ill are one of the most complex subsets of this population. Their special needs, risk factors, and diminished resources create a demand for comprehensive and creative approaches to care management. The purpose of this article is to explore the needs of the critically ill elderly and the role of nursing case management in meeting these needs. A case study approach is used to illustrate typical needs, interventions, and outcomes. PMID- 9392721 TI - Hospice volunteers: a push back to the future. PMID- 9392723 TI - Going one step further: skilled pain assessment and the art of adjuvant analgesia. AB - Evolving pain management practice has two phases: 1) the development of confidence and competence in prescribing opioids at whatever dose is needed to control pain; and, 2) advanced assessment skills and understanding when adjuvant analgesics must be employed with or without opioids to manage certain types of pain. A growing reliance on opioids alone is illustrated in case study by a nurse pain consultant. Dramatic improvements occur in patient function and coping when clinicians move beyond the sole utilization of opioids to understand the importance of skillful assessment of the patient's experience and the artful use of adjuvant medications. PMID- 9392722 TI - Comfort care: a framework for hospice nursing. AB - Provision of comfort is paramount to the practice of hospice nurses. However, the approach to meeting needs holistically is often intuitive or based on multidisciplinary rather than nursing models. A review of the nursing literature identified only one article describing the application of a nursing framework to hospice nursing practice. The purpose of this article is to describe a theory of comfort care that offers definitions and a grid for the art of comfort care that are relevant to hospice nursing practice. Using Kolcaba's framework of holistic comfort, nurses can be comprehensive and consistent in assessing comfort and in designing interventions to enhance the comfort of patients and families. The content domain of holistic comfort is conceptualized as interrelated parts (types and contexts) as they are experienced simultaneously. The framework of comfort care, which includes, the content domain and the theory of comfort, is explained and applied through the presentation of a hospice case study. Potential application of the framework to hospice research is proposed. PMID- 9392724 TI - Advanced media training for hospice CEOs and communication directors. PMID- 9392725 TI - The family with AIDS: multiple challenges for caregivers. AB - Acquired Immune Deficiency Syndrome (AIDS) continues to increase in the United States, especially among minority groups. The disease is impacting not only individuals, but entire families through multiple infections of family members. This results in simultaneous illness and multiple loss within the family system. Delivery and management of quality care for the family is often made difficult by the lack of resources experienced by families with limited income and the multiplicity of problems associated with poverty in the US. This presents numerous challenges to health care providers. Utilizing a case study, this article presents a model demonstrating the coordination of services among several providers as a means of meeting a variety of family needs and providing quality, cost-effective care. PMID- 9392726 TI - Malignant pleural effusions: a brief synopsis. PMID- 9392727 TI - Low-tech nursing guidelines for hospice patients with degenerative neurological disease. PMID- 9392728 TI - Resuscitating and transforming hospice volunteer services. AB - This article is designed for hospice leaders and volunteers to explore creative ways to deal with the stagnancy that can occur in volunteer programs. Specific strategies, such as nurturing caregivers, transitioning into a gift-based program, and integrating spirituality and creativity into volunteer efforts, provide the focus of the article. PMID- 9392729 TI - Gabapentin (Neurontin, Parke-Davis). PMID- 9392730 TI - Hospice and/or palliative care? PMID- 9392731 TI - Nurses do make a difference: but we need to document it! PMID- 9392732 TI - The Pew Commission Report: nursing's challenge to address it. AB - The Pew Health Professions Commission was established in the Spring of 1989 and is administered by the University of California at San Francisco, Center for the Health Professionals. The Commission is charged with assisting health professionals, workforce policy makers, and educational institutions in responding to the challenges of the changing health care system. The Commission's work will identify and explore how regulations protect the public's health, and will propose a new approach to health professionals' licensure, certification, and regulations. In December 1995, the Commission released a full report, Reforming Health Care Workforce Regulation: Policy Considerations for the 21st Century, including 10 recommendations. These recommendations are found in Table 1. In her presentation made during ANNA's 28th National Symposium in Minneapolis on April 28, 1997, Barbara Donaho--a Commissioner on the Pew Health Professions Commission--challenged nephrology nurses to address issues brought up in the Commission Report. The fundamental concepts in Ms. Donaho's keynote presentation were: Articulate the issues that are critical to nurses' licensure as reform is considered. Describe public and professional forces of change if licensure is going to protect the public. Discuss the alternatives that may need to be considered when reform is undertaken by the states to ensure that professionals may practice nursing throughout the country without regulatory and licensing barriers. In this article, based on her ANNA keynote presentation, Ms. Donaho makes it clear that nursing's response to proposed health care reform must be aggressive and swift. PMID- 9392733 TI - ANNA's brief of the response to the report of the taskforce on health care workforce regulation. AB - In December 1995 the Pew Health Professions Commission, a program of the Pew Charitable Trust, released its report titled "Reforming Health Care Workforce Regulations: Policy Considerations for the 21st Century." One of ANNA's external projects for the 1996-97 year was responding to the Commission's report. Western Region Vice President Christine Mudge was selected to serve as project director. In consultation with President Christy Price, she spearheaded ANNA's formal response. Each of the 10 recommendations was assigned to six to eight ANNA members, plus everyone was invited to comment on any portion of the report that they chose. Letters of request for participation were mailed to 66 nephrology nurses. The Board of Directors, committee chairpersons, past ANNA leaders, ANNA consultants, and members at large were involved. The response rate was 59%, or 39 thoughtful critiques of the Commission's recommendations. As project director, Christine Mudge analyzed all responses and formed a draft document. Every effort was made to include all concerns and issues raised by the ANNA participants. The draft document was reviewed at the November ANNA Board of Directors meeting and accepted with some editorial changes. ANNA's response is a 35-page document. Excerpts from ANNA's full response are included on the following two pages. Any ANNA member who desires to receive a copy of ANNA's full response to the Pew Health Professions Commission report may request a copy from the ANNA National Office by calling (609) 256-2320. The Pew Commission is now in the process of reviewing all responses and recommendations to its report. Stay informed by following the ongoing story in the ANNA Update. PMID- 9392734 TI - Evaluation of the pediatric renal transplant recipient. AB - Renal transplantation is the preferred therapy for children with end stage renal disease. A functioning renal allograft can dramatically improve a child's quality of life. Advances in immunosuppressive therapy and clinical practice approaches have significantly improved graft survival rates allowing children to attain near normal growth and development. Accurate assessment and appropriate nursing care enhances long-term survival of these young patients. PMID- 9392735 TI - Subjective burden and quality of life in family caregivers of patients with end stage renal disease. AB - OBJECTIVE: The purpose of this study was to determine the quality of life (QoL) and level of subjective burden reported by family caregivers of persons with ESRD and to examine the relationship between these variables. The influence of patient (gender), illness (dialysis and diabetes status), and caregiver (race, gender, employment status, relationship to patient, and self-rated health) factors on burden and QoL was also examined. DESIGN: An exploratory descriptive design was used. SAMPLE: The convenience sample consisted of 96 caregivers of 96 transplant candidates diagnosed with end-stage renal disease. Participants were recruited from a University transplant service located in the Mid-South. METHODS: Caregivers of patients attending pretransplant clinic evaluations were invited to participate in the study. Caregivers completed a demographic data form, the Caregiver Burden Interview, and General QoL measure. Patient demographic data and dialysis and diabetes status were retrieved from the patient health history database of an ongoing study conducted by our transplant research team. Data were analyzed using descriptive statistics, Spearman's correlation analysis, and the appropriate parametric and nonparametric tests of group differences. RESULTS: Caregivers, most of whom were women, reported good QoL and little to no burden. Caregiver QoL was significantly related to caregiver burden and caregiver self rated health. Neither caregiver race, gender, relationship to the patient, nor patient gender significantly contributed to caregiver burden or caregiver QoL. Caregiver burden did not differ by dialysis type (CAPD, incenter hemodialysis, etc.) or employment category (full-time, part-time, etc.), however QoL differed by employment status. CONCLUSIONS: Findings document the linkages among burden, QoL, and self-rated health as well as illness factors, such as diabetes status. Knowledge about these relationships may facilitate the development of interventions that enhance patient and family outcomes. PMID- 9392737 TI - Vision screening in older adults on dialysis: do nephrology nurses have a role? AB - Undetected vision loss commonly occurs in older adults adding undue stress to those on dialysis. Poor vision is associated with increased risk of falls and decreased quality of life. Common visual impairments--presbyopia, cataracts, age related macular degeneration, glaucoma, and diabetic retinopathy--often are detectable by visual acuity testing. Using various methods of visual acuity testing, nephrology nurses can perform vision testing quickly and inexpensively. Other nursing interventions also can improve eyesight. PMID- 9392738 TI - Pulse dose oral calcitriol therapy for renal osteodystrophy: literature review and practice recommendations. AB - Oral pulse dosing of calcitriol has been proposed as an alternative to intravenous administration in the treatment of renal osteodystrophy. A review of the literature suggests it can provide a safe and effective method that is especially convenient for peritoneal dialysis patients with mild to moderate bone disease. However, its use should be accompanied by careful monitoring and control of serum calcium, phosphorus, and parathyroid hormone levels. Some practical suggestions are provided. PMID- 9392739 TI - Flipping or rotating fistula needles: readers' responses. PMID- 9392740 TI - Epoetin alfa: focus on maintaining a higher, stable, Hct. Case study of the anemic patient. AB - Clinical evidence indicates that maintaining a stable hematocrit (Hct) higher in the target range of 30% to 36% can lead to improvement in overall patient outcomes. On the basis of these data, a recent analysis by the Dialysis Outcomes Quality Initiative Anemia Work Group has recommended a target Hct of 33% to 36% (hemoglobin 11 g/dl to 12 g/dl). Maintaining a stable Hct higher in the target range provides nurses and other dialysis clinicians with two benefits: improved patient care and decreased time and costs for patient management. This article focuses on the data supporting such a policy. Clinical practices from two prominent dialysis centers are presented as models of good anemia management. PMID- 9392741 TI - Authors' response: no increased risk for peritonitis by insulin injected into the peritoneal dialysis solution bag through Coloplast. PMID- 9392742 TI - Implementing innovation. PMID- 9392743 TI - Protecting our youngest girls. PMID- 9392744 TI - Are you diagnosing domestic violence? PMID- 9392745 TI - Preventing osteoporosis. PMID- 9392746 TI - Emergency contraception. The pill's little-known secret goes public. PMID- 9392747 TI - On your markers. Research advances in breast cancer in women. PMID- 9392748 TI - News from the front. This nurse soldier is waging war on breast cancer. PMID- 9392749 TI - Fluid check. Making the case for intrapartum amnioinfusion. PMID- 9392750 TI - Shark-proof your practice. How the law is every nurse's best ally. PMID- 9392751 TI - Taking time to care. Making ways to help teen parents. PMID- 9392752 TI - Recognizing and preventing antibiotic-associated complications in the critical care setting. AB - Antibiotics are commonly used, even overused, in the intensive care unit (ICU) patient population. These agents, though useful and often lifesaving, are associated with many toxicities. These include infusion-related problems, allergic reactions, organ-system toxicities, drug accumulation in patients with renal disease, drug interactions, antibiotic resistance, diarrhea, interference with laboratory tests, and more. Informed vigilance by the ICU nurse can be helpful in increasing the benefit/toxicity ratio associated with these agents. PMID- 9392753 TI - Antifungals: use in high-risk patients. AB - Advanced medical technology has resulted in a growing population of patients with altered defense mechanisms against nosocomial infections. Fungal infections, primarily Candida species, account for a significant proportion of hospital acquired infections and are associated with increased morbidity and mortality. This article discusses current issues and controversies related to nosocomial fungal infections and summarizes optimal management strategies. PMID- 9392754 TI - Antimicrobial resistance: implications for the clinician. AB - Antimicrobial resistance has become an issue of global proportion, and this dilemma has greatly affected the intensive care unit (ICU) setting. Increased antimicrobial use and other selective pressures have created an environment in the ICU that is a testing ground for survival of microorganisms. By various mechanisms, gram-positive cocci and gram-negative bacilli are becoming more resistant to our current armamentarium of antimicrobials. Strategies to improve the current situation of reduced microbial susceptibility include the following: continued and specific surveillance in ICUs, antimicrobial protocols, continued isolation precautions and appropriate handwashing, increased education at all levels, increased use of immunotherapies, better use of our current antimicrobial agents, and increased research in finding new and innovative antimicrobial agents while curtailing the marketing ploys that promote excessive and inappropriate use of these valuable medications. PMID- 9392755 TI - Hospital-acquired pneumonia and its management. AB - Hospital-acquired pneumonia (HAP) is an important cause of morbidity and mortality in the United States. Classifying the patient's pneumonia by the presence or absence of risk factors helps determine what organisms need to be considered as etiologic agents so that empiric antibiotic therapy can be initiated while cultures are pending. Medical care personnel can also use preventive strategies to help decrease the incidence of nosocomial pneumonia. This article will discuss pathogenesis, diagnosis, management, and prevention of HAP. PMID- 9392756 TI - The microbiology laboratory's role in life-threatening infections. AB - The microbiology laboratory's contribution to the care of patients who are critically ill is explored. The economic and epidemiologic impact of broad spectrum antibiotic use is discussed, along with methodology of antimicrobial susceptibility testing and the potential value of an antibiogram. The clinical benefits of blood, sputum, urine, spinal fluid, and wound cultures are discussed, with an emphasis on interpretation of culture results. The significance of, and procedures for, proper specimen collection are emphasized throughout the article, and the ramifications of improper specimen collection are presented. PMID- 9392757 TI - Antibiotics: why they fail in patients who are critically ill. AB - Most of the failures of antimicrobial therapy can be related to advanced infections in patients with serious comorbid processes or altered immunity. However, some of the failures are related to the type of antimicrobial therapy used, the length of therapy, the pharmacokinetic or pharmacodynamic properties of the antimicrobial agent, the development of antimicrobial resistance, microbial factors that influence antimicrobial efficacy, and the site and type of infection. This report will review the common mechanisms by which antimicrobials fail. PMID- 9392759 TI - Prevention of infections related to central venous catheters. AB - Infections related to central venous catheters (CVCs) increase hospital costs, length of stay, and patient mortality. Review of the literature and research pertaining to CVCs have provided some guidelines to reduce the risk of infections related to CVCs. Recommended guidelines include use of sterile technique with insertion, maintenance of site dressings, avoidance of routine changes of CVCs, and reduction of hub manipulation. Critical care nurses have the primary opportunity to improve patient outcomes by reducing CVC infections. PMID- 9392758 TI - Management of wounds and wound infections in the intensive care unit. AB - Many factors combine to make management of wounds and wound infections in patients in intensive care units (ICUs) a complex task. An understanding of the anatomy, pathophysiology, and bacteriology provides a framework to approach these patients. The patient's underlying disease influences the care of the wound. Wound factors such as necrotic tissue, bacterial load, or presence of fistulae or a foreign body have important impact on the patient's care. With assessment and knowledge of normal healing, timely intervention in the ICU can identify patients whose wounds are not healing properly and allow for corrective interventions to help the patient return to normal function. PMID- 9392761 TI - New treatment options for psoriasis. AB - Psoriasis is a chronic genetic disease characterized by hyperproliferation and inflammation of the epidermis and dermis. Presently there is no cure for psoriasis, only treatments to alleviate the symptoms. Tazarotene (Tazorac) is a new vitamin A derivative and the first topical retinoid to demonstrate therapeutic success in treating plaque-type psoriasis. This article will introduce you and your patients to the safe and effective use of tazarotene. PMID- 9392762 TI - UVB therapy: dermatology nursing considerations. AB - Each patient responds uniquely to phototherapy treatment. Not every patient can tolerate daily treatment. Different areas of the body require different amounts of light in order to clear. Precise documentation of erythema is important for both treatment and legal reasons. A negative response to UVB therapy can be from either ineffective treatment or an adverse reaction to UVB. PMID- 9392760 TI - Antibiotic classification: implications for drug selection. AB - This article presents a useful antibiotic classification model for the busy clinician who must select agents for patients in the critical care setting. The model organizes antibiotics based on their mechanism of action, ie, cell-wall inhibitors, nucleic acid inhibitors, and protein-synthesis inhibitors; clinicians are encouraged to further segregate agents within the broader categories by generation. An overview of the antimicrobial spectrum for each class is presented, and important differences within individual classes are highlighted. The most common indications for each antibiotic class are reviewed, and key pharmacokinetic characteristics that help distinguish one drug from another are outlined. PMID- 9392763 TI - Facial rejuvenation with botulinum. AB - Botulinum toxin type A (Botox) blocks the release of neurotransmitter acetylcholine at the presynaptic neuromuscular junction leading to an irreversible, but temporary muscular paralysis and weakness. This can produce a significant improvement of wrinkling in the upper face caused by the actions of the facial muscles. A prospective clinical study representing a 15-month experience with this new technique is presented. Patient selection and evaluation, classification of animation lines, techniques, results, and complications are discussed. PMID- 9392764 TI - What's your assessment? Tinea versicolor. PMID- 9392765 TI - Azelaic acid 20% cream (AZELEX) and the medical management of acne vulgaris. AB - Azelaic acid 20% cream (AZELEX) is a novel anti-acne agent with antimicrobial activity and keratinization-normalizing properties. In acne it is broadly comparable in efficacy to 0.05% tretinoin, 5% benzoyl peroxide, and 2% erythromycin, but is less irritating than tretinoin and benzoyl peroxide. PMID- 9392766 TI - Latex as a killer: one nurse's story. PMID- 9392767 TI - "The future of ASORN and ophthalmic nursing: where are we going"? PMID- 9392768 TI - The bigger the crisis, the bigger the opportunity! AB - Every time I pick up the newspaper, a scientific or nursing journal, or more recently, an issue of Time, I find data, complete with startling facts and figures, that pierce my very soul, about the tragedies occurring in our nation's hospitals caused by the restructuring movement attempting to make health care a "bottom line" business. And I know all too well, as do most of you, how undetermined and tentative we are about the future for nurses. PMID- 9392769 TI - Coats' disease. AB - A 3-year-old boy had leukocoria in his right eye that could have been confused for retinoblastoma because leukocoria is the most common presenting sign of retinoblastoma. Educating parents, nurse practitioners, pediatricians, and fellow health care professionals about such presenting ocular signs will help expedite ophthalmic examinations and accurate diagnosis for these children. PMID- 9392770 TI - Ophthalmic side effects of hyperbaric oxygen therapy. AB - Hyperbaric oxygen (HBO) treatments are documented to cause ocular side effects. According to Palmquist et al., HBO therapy has been used for many years, yet there are only a few reports of its effects on the eye. With current studies reporting lens changes, as well as hyperopic and myopic fluctuations, the role of the nurse in assessing and reporting vision changes needs to be defined and clarified. PMID- 9392771 TI - Choroidal osteoma: acoustic shadowing and reduplication echoes. AB - A 27-year-old woman had a curious choroidal mass of 12 years duration in her right eye. Interesting ultrasonic findings of a choroidal osteoma, including acoustic shadowing and reduplication echoes on A-scan and B-scan are presented. Ophthalmic nurses can assist in performing ophthalmic examinations and in reinforcing regular follow-up examinations for these patients. PMID- 9392772 TI - Genetic teaching for the retinoblastoma patient. AB - Retinoblastoma is the most common primary intraocular tumor in children. Retinoblastoma can be hereditary (familial) or nonhereditary (nonfamilial). Nurses who have an understanding of the genetic patterns for retinoblastoma can participate in the counseling of these patients. A chart is provided as a tool for teaching patients about family patterns of retinoblastoma. PMID- 9392773 TI - ASORN member Von Best Whitaker elected to American Academy of Nursing. PMID- 9392774 TI - Career options: the increasing demand for courtroom nurses. AB - Gail Hoffman and Lisa Michaels first met at the University of Miami School of Nursing in the early seventies. Because of their mutual interest in pediatrics and their desire to stay in South Florida, both expected that their paths would cross again at some time during their nursing careers. But neither of them expected that 20 years later the crossroads would be in a courtroom, both working for a defense attorney on a medical malpractice case. Now, in Smith v Mizrahi, Gail is a nurse paralegal assigned to the case through her employment at Green, Williams, & Conrad, P.A., and Lisa was called to serve as an expert witness. Although the coincidence of Gail and Lisa's reacquaintance is unique, their career change stories are not. PMID- 9392775 TI - RNdex Top 100, CD-ROM. PMID- 9392776 TI - Penetrating ocular trauma and visual outcome. PMID- 9392777 TI - Cognitive deficit in 7-year-old children with prenatal exposure to methylmercury. AB - A cohort of 1022 consecutive singleton births was generated during 1986-1987 in the Faroe Islands. Increased methylmercury exposure from maternal consumption of pilot whale meat was indicated by mercury concentrations in cord blood and maternal hair. At approximately 7 years of age, 917 of the children underwent detailed neurobehavioral examination. Neuropsychological tests included Finger Tapping; Hand-Eye Coordination; reaction time on a Continuous Performance Test; Wechsler Intelligence Scale for Children-Revised Digit Spans, Similarities, and Block Designs; Bender Visual Motor Gestalt Test; Boston Naming Test; and California Verbal Learning Test (Children). Clinical examination and neurophysiological testing did not reveal any clear-cut mercury-related abnormalities. However, mercury-related neuropsychological dysfunctions were most pronounced in the domains of language, attention, and memory, and to a lesser extent in visuospatial and motor functions. These associations remained after adjustment for covariates and after exclusion of children with maternal hair mercury concentrations above 10 microgram(s) (50 nmol/g). The effects on brain function associated with prenatal methylmercury exposure therefore appear widespread, and early dysfunction is detectable at exposure levels currently considered safe. PMID- 9392778 TI - Effect of postnatal exposure to a PCB mixture in monkeys on multiple fixed interval-fixed ratio performance. AB - Behavioral impairment as a consequence of PCB exposure beginning in utero has been reported in both humans and animals. The present study assessed the behavioral consequences of postnatal exposure to PCBs. Male monkeys (Macaca fascicularis) were dosed from birth to 20 weeks of age with 7.5 microgram(s)/kg/day of a PCB mixture representative of the PCBs typically found in human breast milk (eight monkeys) or vehicle (four monkeys). At 4 years of age, performance under a multiple fixed interval (FI)-fixed ratio (FR) schedule of reinforcement was assessed. The FI component was more sensitive to disruption as a result of PCB exposure than was the FR component. PCB-exposed monkeys displayed shorter mean interresponse times (IRTs) than controls, particularly during the earlier sessions of the experiment. Similarly, the increase in pause time characteristic of the acquisition of typical FI performance emerged more slowly across sessions in the PCB-treated group. However, the number of short IRTs (less than 5 s) remained greater in the treated group compared to controls over the 48-session duration of the experiment. On the FR component, control monkeys decreases the mean pause time across sessions whereas the PCB-treated group did not; there were no differences between groups for absolute value of average IRT or pause time. The results of this study extend previous research in this cohort of monkeys, and provide further evidence that PCB exposure limited to the early postnatal period and resulting in environmentally relevant body burdens produces long-term behavioral effects. PMID- 9392779 TI - Neonatal binge ethanol exposure using intubation: timing and dose effects on place learning. AB - Neonatal rats were given ethanol using an acute intubation procedure that resulted in daily binge-like exposure with minimal effects on somatic growth. Acquisition of place learning in the Morris water maze was evaluated on postnatal days (PD) 26-31. In Experiment 1, a total of 5.25 g/kg/day of ethanol was administered in two daily intubations on PD 4-6, PD 7-9, or PD 4-9, producing mean peak BACs of 265 mg/dL. Place learning acquisition deficits in a 114-cm diameter tank were found for the PD 4-9 and PD 7-9 groups, but not the PD 4-6 group. In Experiment 2, either 4.5 or 5.25 g/kg/day of ethanol was administered on PD 7-9 and place learning was tested in a 171-cm-diameter tank. Significant acquisition deficits resulted from the higher dose, and probe trial search patterns for both ethanol groups were significantly less localized than controls. In Experiment 3, no significant effects of either PD 7-9 dose were found on a visible platform task. These findings reveal selective place learning deficits in this intubation model of neonatal binge exposure, and confirm a temporal window of vulnerability to spatial learning deficits during the second neonatal week. PMID- 9392780 TI - Evaluation of tremor in aluminum production workers. AB - A cross-sectional study of 63 current and former aluminum potroom workers and 37 comparison workers was conducted to evaluate for evidence of neurological dysfunction, including tremor from long-term exposures to aluminum using sensitive quantitative measures of arm/hand and leg tremor. Signs of upper extremity tremor were also evaluated by neurological examination and compared with the quantitative measures of arm/hand tremor. Both arm/hand and leg tremor were measured using fatiguing test conditions, but no statistically significant differences due to exposure to aluminum were present between the potroom workers and the comparison workers. The neurological examination also showed no statistically significant differences between the groups on the evaluation of signs of tremor. These results do not support the findings of Best-Pettersen et al., who reported evidence of increased tremor in aluminum workers using the static steadiness test in the Halstead-Reitan battery. Differences between the studies that may have contributed to the contrasting results are discussed. In addition, techniques are presented for using microcomputer-controlled devices to evaluate tremor in both the visible (1-6 Hz) and nonvisible (7-18 Hz) frequencies of the tremor spectrum. PMID- 9392781 TI - Critical issues in the use and analysis of the Lanthony Desaturate Color Vision test. AB - The Lanthony Desaturate Color Vision test (D-15d) has been used to demonstrate the incidence of acquired color vision defects resulting from toxic exposure. The D-15d is a sensitive test designed to grade color deficiencies, but results can be difficult to interpret beyond the qualitative level, and the high incidence of errors reported for controls in some toxicology studies raises questions about how to effectively use this test. This article reviews standard administration of the test, physical determinants of performance, classification of acquired color vision defects, and methods of analysis that have been used to quantify results. The basis for a new method of analysis is discussed, illustrating the source of some characteristic errors, and recommendations are made for test protocols to attempt to more closely identify the type of color vision loss with the goal of identifying the site of toxicological insult. PMID- 9392782 TI - Prenatal nicotine sex-dependently alters agonist-induced locomotion and stereotypy. AB - This study examined the effects of prenatal nicotine exposure (2 mg/kg/day) via s.c. osmotic minipumps, gestational days 7-22, on nicotine- and lobeline-induced locomotor activity and stereotypy in 14-day-old rat pups. Prenatal nicotine exposure increased fetal mortality and produced decreases in weight gain apparent after weaning, but did not affect acquisition of developmental milestones. Compared to male pups prenatally exposed to saline, those prenatally exposed to nicotine and challenged with nicotine (1 mg/kg, i.p.) exhibited significantly greater locomotor activity, whereas a lobeline challenge (1 mg/kg, s.c.) produced significantly greater stereotypy. No effects of prenatal exposure were observed on locomotor activity or stereotypy in females. Results suggest that 1) central control of motor function may be more vulnerable to prenatal nicotine in males, and 2) nicotine and lobeline possess distinct pharmacological profiles. PMID- 9392783 TI - Perinatal delta(9)-tetrahydrocannabinol exposure alters the responsiveness of hypothalamic dopaminergic neurons to dopamine-acting drugs in adult rats. AB - We have recently reported that perinatal cannabinoid exposure altered the normal development of dopaminergic neurons in the medial basal hypothalamus at early postnatal and peripubertal ages. Most of these effects tended to disappear in adulthood, although we suspect the existence of a persistent, but possibly silent, alteration in the adult activity of these neurons. To further explore this possibility, we evaluated the responsiveness of these neurons to pharmacological challenges with a variety of dopaminergic drugs administered to adult male and female rats that had been exposed to delta(9)-tetrahydrocannabinol (delta(9)-THC) or vehicle during the perinatal period. In the first experiment, we evaluated the sensitivity of hypothalamic dopaminergic neurons to amphetamine (AMPH), which causes enhancement of dopaminergic activity by a variety of mechanisms. The most interesting observation was that both adult males and females, when perinatally exposed to delta(9)-THC, showed a more marked AMPH induced decrease in the production of L-3,4-dihydroxyphenylacetic acid (DOPAC), the main intraneuronal metabolite of dopamine (DA), although this did not affect the prolactin (PRL) release. In the second experiment, we evaluated the in vivo synthesis of DA by analyzing the magnitude of L-3,4-dihydroxyphenylalanine (L DOPA) accumulation caused by the blockade of L-DOPA decarboxylase with NSD 1015. As expected, NSD 1015 increased L-DOPA accumulation and decreased DOPAC production, with a parallel increase in PRL release, all of similar magnitude in both delta(9)-THC- and oil-exposed adult animals. In the last experiment, we tested the magnitude of the increase in PRL release produced by the administration of either SKF 38393, a specific D1 agonist, or sulpiride, a specific D2 antagonist. Both compounds increased plasma PRL levels in adult animals of both sexes, the effects in females being significantly more marked. The perinatal exposure to delta(9)-THC also modified the degree of increase in plasma PRL levels induced by both compounds, with opposite responses as a function of sex. Thus, delta(9)-THC-exposed females responded more intensely to SKF 38393 and, particularly, to sulpiride than oil-exposed females, whereas delta(9)-THC-exposed males responded to SKF 38393 lesser than oil-exposed males, although both responded equally to sulpiride. In summary, our results are consistent with the possible existence of subtle changes in the activity of hypothalamic dopaminergic neurons in adulthood caused by the exposure to delta(9) THC during perinatal development. These silent changes could be revealed after the administration of drugs such as: (i) AMPH, whose effect producing a decreased DOPAC accumulation was more marked in delta(9)-THC-exposed males and females; and (ii) SKF 38393 and sulpiride, whose stimulatory effects on PRL secretion were of different magnitude in delta(9)-THC-exposed animals, with an evident sexual dimorphism in the response. The neurochemical basis for these differences remains to be determined. PMID- 9392785 TI - Effects of neonatal naltrexone on neurological and somatic development in rats of both genders. AB - The effects of a daily injection of the opioid antagonist naltrexone (NALTX, 1 mg/kg, s.c.) from birth to weaning on various parameters were investigated in male and female rats during postnatal development until adulthood. NALTX increased cerebellar DNA and protein content as well as cerebellar weight at 7 days. Eye opening was not affected by NALTX but it appeared advanced by 1 day in all groups compared to other studies, possibly due to a handling effect caused by the daily injection. Water and food consumption were augmented by NALTX during days 23-32 and days 55-70. Treated preweaning animals showed lower body growth rates than controls. However, NALTX caused a moderate increase in body weight measured during postweaning until adulthood. The effects of NALTX on the parameters evaluated (excepting the cerebellar measurements on day 7), although clearly statistically significant, were small in absolute terms. The preweaning opioid blockade caused by our NALTX treatment seems to affect more markedly the neural and behavioural development than the somatic growth. This work also provides potentially useful baseline data for the study of male and female rats during postnatal development. PMID- 9392784 TI - Effects of lead-arsenic combined exposure on central monoaminergic systems. AB - Lead acetate (116 mg/kg/day), arsenic (11 or 13.8 mg/kg/day as sodium arsenite), a lead-arsenic mixture or vehicle were administered to adult mice through gastric intubation during 14 days. Then, the regional content of norepinephrine (NE), dopamine (DA), serotonin (5-HT), 3,4 dihydroxyphenyl-acetic acid (DOPAC), 5 hydroxyindole-3-acetic acid (5-HIAA), arsenic, and lead were quantified. Compared with the accumulation after single element exposures, the mixture elicited a higher accumulation of lead and a lower arsenic accumulation in the brain. Compared to controls, lead induced only an augmentation of DOPAC (200%) in the hypothalamus. By contrast, the mixture provoked increases of DOPAC in the hypothalamus (250%), DA and 5-HIAA in the striatum (67 and 187%, respectively) and NE decreased in the hypothalamus (45%). Although these alterations were similar to those produced by arsenic alone, the mixture provoked a 38% decrease of NE in the hippocampus and increases of 5-HT in midbrain and frontal cortex (100 and 90%, respectively) over control values, alterations that were not elicited by either metal alone. These results demonstrate an interaction arsenic/lead on the central monoaminergic systems of the adult mouse. PMID- 9392786 TI - Prenatal exposure of rats to diphenhydramine: effects on physical development, open field, and gonadal hormone levels in adults. AB - Diphenhydramine (DPH), a classical H1 receptor antagonist, has been used in pregnancy for the treatment of allergies, nausea, and vomiting. It has been reported that 10-20% of pregnant women take antihistamine-containing preparations at some point during pregnancy. The present study analyzed the influence of prenatal exposure to DPH of rats on: 1) maternal behavior and milk production of dams; 2) physical and reflexologic development of offspring; and 3) long-term effects on open field behaviors and gonadal hormone levels in offspring. Female pregnant rats were injected s.c., daily, with 20 mg/kg DPH or saline from embryonic day (E) 0 to 21. After delivery, maternal behavior was assessed and offspring physical and reflexologic development was examined. Open field activity of male and female rats was measured at 21 and 75 days of age and plasma hormone levels were evaluated in both sexes at 120 days of age. Neither maternal behavior nor milk production was affected by DPH treatment. Treated offspring showed an accelerated pinna unfolding, eye opening, and a delay of testes descent and vaginal opening. Both righting reflex and negative geotaxis development were accelerated, but prenatal exposure to DPH did not modify offspring locomotor activity. When tested as adults, a lack of sexual dimorphism in the open field activity of males and females was observed. No differences were observed between gonadal hormone levels of control and experimental groups of either sex. The findings suggest that prenatal DPH exposure influences physical and reflex development of rat pups. PMID- 9392787 TI - Susceptibility of adult rats to lead-induced changes in NMDA receptor complex function. AB - Because cognitive impairments can occur with occupational Pb exposures, changes in NMDA receptor complex function might be expected to occur in adult rats treated with Pb. Using drug discrimination procedures, MK-801 sensitivity was determined in adult rats at three time points: after chronic exposure to 0, 50, or 150 ppm Pb acetate; again after exposure levels were increased to 500 and 1000 ppm; and 6 months after termination of Pb exposure. Changes in blood (PbB) and brain Pb levels, and in MK-801 and CGP-39653 binding, were examined in additional groups of nonbehaviorally tested rats. Pb decreased MK-801 sensitivity after Pb exposure levels were increased, but only at 500 ppm, indicating biphasic effects and precluding any correspondence between behavioral changes and biomarkers of exposure. Associated PbBs were higher, but brain Pbs similar to those associated with MK-801 sensitivity changes following postnatal and postweaning exposures. Neither MK-801 nor CGP-39653 binding was systematically affected by chronic Pb exposure in adults. Although adult Pb exposures do produce changes in NMDA function, at least as indicated by changes in MK-801 sensitivity, vulnerability to such effects is clearly less pronounced than with exposures occurring earlier in development. PMID- 9392788 TI - Not all DMSA defects are scars. PMID- 9392789 TI - Diagnostic usefulness of lung SPET in pulmonary thromboembolism: an outcome study. AB - The lung single photon emission tomographic (SPET) images of 985 consecutive patients referred for suspected pulmonary embolism were correlated with clinical outcome and angiography to evaluate the clinical usefulness of lung SPET compared to conventional planar ventilation/perfusion lung imaging. SPET interpretations followed the revised PIOPED criteria and clinical outcome was determined from referring physicians, hospital records, direct patient contact and county hall records. Patients were deemed to have had no clinically significant pulmonary embolism at the time of the SPET examination if, within the following 3 months: (1) the patient was alive and had no clinical evidence of pulmonary embolism or, (2) if deceased, pulmonary embolism was unlikely to have been the cause of death. Operating characteristics were based on the methods of Choi and of Simel. SPET interpretation was categorized as follows: high probability, 143 (14%); low probability, 840 (82%); intermediate, 41 (4%) (in contrast to PIOPED, with 39% intermediate interpretations). Pulmonary angiography was performed in only 4% of patients. Adequate follow-up data were available for 97% of patients. To facilitate comparison with PIOPED, either a high-probability or an intermediate probability or an intermediate-probability study was considered to be a positive test, and either a low-probability or a normal study was considered to be a negative test. The sensitivity was 83% (PIOPED 82%), specificity 92% (PIOPED 52%), positive predictive value 62% (PIOPED 47%) and negative predictive value 97% (PIOPED 85%). The positive and negative predictive values have not been corrected for prevalence, which was approximately twice as high in the PIOPED study. Lung SPET provided accurate diagnostic information in 96% of patients and specificity was greatly improved compared to planar lung imaging reported in PIOPED. The diminished need for angiography greatly reduced the cost of evaluating patients suspected of having pulmonary embolism. PMID- 9392790 TI - Pentavalent 99Tcm-DMSA imaging in patients with bone metastases. AB - Pentavalent 99Tcm-dimercaptosuccinic acid (99Tcm-(V)DMSA) has an established role in imaging medullary thyroid carcinoma. There have been case reports of uptake in bone metastases. Our aims were to compare 99Tcm-(V)DMSA with 99Tcm hydroxymethylene diphosphonate (99Tcm-HDP) in bone metastases, to assess its value in imaging of bone metastases, and to assess the prospects of the beta emitting analogues 186/188Re-(V)DMSA as palliative agents for painful bone metastases. Ten patients confirmed by a 99Tcm-HDP bone scan to have bone metastases secondary to carcinoma of the prostate, lung or breast were injected with 99Tcm-(V)DMSA (600 MBq). Whole-body scans acquired at 3 and 24 h were compared with the 99Tcm-HDP bone scans. 99Tcm-(V)DMSA showed high soft tissue background, kidney retention and avid uptake in most bone metastases: 86% of bone lesions identified on bone scans were detected with 99Tcm-(V)DMSA. The lesion-to normal ratios were comparable to or lower than those for 99Tcm-HDP at 3 h, but increased by 24 h. Instances of abnormal uptake in liver, primary lung tumour, lymph nodes and pleural effusion were observed. We conclude that 99Tcm-(V)DMSA is a tracer for bone metastases (with lower sensitivity than 99Tcm-HDP) and soft tissue tumours. If 186/188Re-(V)DMSA behave similarly, they may find use in therapy for soft tissue tumours and bony metastases. PMID- 9392791 TI - How does gated SPET alter reporting of myocardial perfusion studies? AB - Gated single photon emission tomography (SPET) during myocardial perfusion scintigraphy has been proposed as a method for distinguishing artefact from myocardial infarction and for assessing myocardial viability. This study describes the alterations in the specialist report produced when gated SPET was included in 50 consecutive myocardial perfusion studies. Diagnostic confidence was scored following an initial assessment using non-gated images alone and then re-scored after a review of the gated SPET data. The change in diagnostic confidence and the presence of additional information about myocardial viability were recorded. These were correlated with various clinical parameters, including a subjective assessment of the likelihood of an attenuation artefact as determined by clinical examination prior to imaging. Diagnostic confidence was altered by identification of attenuation artefacts in 11 (22%) patients, moving towards normality in all cases. In three of these (6%), the change was sufficient to make coronary angiography potentially unnecessary. Attenuation artefacts were equally common in men and women. Whereas breast attenuation artefacts could be anticipated from clinical examination, this was not so far diaphragmatic attenuation artefacts, which occurred most commonly in men under 55 years of age. The gated study provided additional information about myocardial viability in 13 (26%) patients, most commonly those referred following myocardial infarction. In view of the frequent alterations in the specialist's final report and the difficulties in pre-selecting patients likely to benefit from the technique, the use of gated SPET can be justified for all patients undergoing sestamibi myocardial perfusion imaging. PMID- 9392792 TI - The effect of training on the interpretation of 99Tcm-sestamibi myocardial perfusion SPET in patients with suspected coronary artery disease. AB - The aim of this study was to examine the effect of a period of concentrated training in nuclear cardiology on the accuracy of reporting 99Tcm-sestamibi (99Tcm-MIBI) single photon emission tomographic (SPET) images. Two visiting cardiologists, with no previous experience in nuclear cardiology, were asked to report blindly 60 99Tcm-MIBI SPET scans after 2 weeks of training in nuclear cardiology. One (observer 2) reported the same scans blindly after 2 months of further training. The results were compared with the assessment made by two experienced nuclear cardiologists and by using kappa statistics. Kappa values for the overall interpretation of the scan (normal or abnormal), segmental analysis (normal, ischaemic, fixed or mixed) and the three arterial territories were 0.7, 0.58 and 0.67 respectively. Following 2 months of further intensive training of observer 2, the kappa values were 0.857, 0.78 and 0.91 respectively. The difference between the two readings of observer 2 was significantly different for the segmental analysis (P < 0.001) and arterial territories (P = 0.006) but it did not reach statistical significance for the overall interpretation (P = 0.7). Thus, cardiologists without previous interpretation skills in nuclear cardiology required about 2 months of intensive training to achieve good accuracy in the interpretation of 99Tcm-MIBI SPET images. Accordingly, these techniques can be established in centres other than tertiary sites. PMID- 9392794 TI - Hydration does not influence the image quality in bone scintigraphy: an investigation using 99Tcm-HDP. AB - We investigated the possibility that the increased diuresis caused by hydration leads to enhanced renal excretion of radiotracer and thereby increases the target to-background activity ratio and improves the image quality in bone scintigraphy. The study was carried out using paired comparisons in 10 healthy volunteers. Whole-body antero-posterior and postero-anterior acquisitions were obtained for 4 h after the administration of 99Tcm-hydroxymethylene diphosphonate with and without hydration. The bone-to-soft tissue activity ratio was semi-quantified by comparing regions of interest acquired from geometrical means of the acquisitions. Hydration was performed by slowly drinking 1.5 litres of water after administration of the tracer. Hydration induced a slight enhancement of excretion of the activity, but it had no effect on the bone-to-soft tissue activity ratio, nor did it influence image quality. We conclude that hydration could be avoided when it is cumbersome for the patient. However, as hydration reduces the radiation dose to the urinary bladder wall, which is the critical organ, hydration should be maintained in younger patients. PMID- 9392793 TI - 99Tcm-MDP blood-pool phase in the assessment of repetitive strain injury. AB - We reviewed three-phase bone scans of the limbs of 7 patients suffering from limb pain suggestive of occupational repetitive strain injury (RSI) and compared them with 13 patients with limb pain due to various aetiologies. Doppler ultrasound measurement of blood flow had been performed in 13 of the 20 patients. The bone scan results showed increased blood flow and pooling (second phase) in the affected limbs of patients with RSI as compared to those with algodystrophy or non-specific limb pain (sensitivity 86%, specificity 85%). Doppler ultrasound also demonstrated increased blood flow to the affected limbs (sensitivity 83%) but failed to differentiate between the different aetiologies of pain (specificity 14%). We conclude that the blood-pool phase of three-phase bone scans can play a potential role in screening RSI patients. PMID- 9392795 TI - Functional evaluation of the remaining kidney in kidney donors by radionuclide dynamic imaging using a graphic method with factor analysis. AB - An uptake coefficient proportional to the glomerular filtration rate (GFR) can be estimated from dynamic renal images with 99Tcm-DTPA using Rutland's graphic method. We have developed a new method of extracting the input and retention functions by applying factor analysis to the renal dynamic images obtained with 99Tcm-DTPA, which we have called the 'factor uptake coefficient' (Factor UC). In the present study, we followed 13 living kidney donors (7 males, 6 females) by measuring the Factor UC in each kidney before nephrectomy and in the remaining kidney 3 weeks and 1 year after nephrectomy. The median Factor UC in the remaining kidney increased from a pre-nephrectomy value of 0.31 to 0.52 three weeks post-nephrectomy, which then remained unchanged for up to 1 year. These results indicate that functional compensation occurs following unilateral nephrectomy, and that this process is complete within 3 weeks after nephrectomy. PMID- 9392796 TI - Visualization of frontal postural hypoperfusion in patients with Takayasu arteritis with upright 99Tcm-HMPAO brain SPET. AB - Takayasu arteritis is a chronic inflammatory angiopathy involving the cerebral arteries. We performed upright and supine 99Tcm-HMPAO brain single photon emission tomography (SPET) to investigate the cerebral perfusion pattern in eight patients with Takayasu arteritis, and we compared the results with those acquired using 123I-IMP and acetazolamide in six patients. SPET images were evaluated visually and semi-quantitatively. Hypoperfusion was visually detected in all eight patients during the provocative upright test with 99Tcm-HMPAO, and in three of six tested using acetazolamide and 123I-IMP. Semiquantitative analysis revealed that the mean cortical-to-cerebellar ratio in the upright position was significantly changed compared to that in the supine position in the right frontal area (from 0.86 +/- 0.07 to 0.91 +/- 0.09; P < 0.05). Change was also seen in the left frontal area (from 0.85 +/- 0.08 to 0.91 +/- 0.08; P < 0.05). No significant change was seen in other cortical areas with the upright test or in any areas with the acetazolamide test. We postulate that reduced arterial compliance may cause frontal postural hypoperfusion in patients with Takayasu arteritis due to poor functioning of autoregulation and arterial stenosis or occlusion. We conclude that the provocative upright test with 99Tcm-HMPAO brain SPET can detect abnormal patterns of cerebral perfusion in patients with Takayasu arteritis that might be missed by brain SPET using 123I-IMP and acetazolamide. PMID- 9392797 TI - Scintigraphic varieties in Hashimoto's thyroiditis and comparison with ultrasonography. AB - The scintigraphic findings in Hashimoto's thyroiditis are highly variable and can mimic any thyroid abnormality. In this study, we compared the scintigraphic findings with ultrasonography in 48 patients with Hashimoto's thyroiditis. Thyroid scintigrams revealed diffuse hyperplasia in 12 patients, multinodular goiter in 20 patients and a solitary nodule in 16 patients (toxic adenoma, n = 1; hypoactive nodule, n = 4; hyperactive nodule with no suppression, n = 3; normoactive nodule, n = 8). Ultrasonography revealed diffuse hyperplasia in 19 patients, multinodular goiter in 20 patients and a solitary nodule in 9 patients. The thyroid scan and ultrasonography revealed the same findings of diffuse hyperplasia in 12 patients and multinodular goiter in 20 patients. Of the 16 patients with a solitary nodule on scintigraphy, only 9 showed the same finding on ultrasonography, with the other 7 showing diffuse hyperplasia. The difference in nodularity between thyroid scanning (74.9%) and sonography (60.4%) has been attributed to pseudonodularity in Hashimoto's thyroiditis. In conclusion, our results confirmed that Hashimoto's thyroiditis can mimic any thyroid abnormality, including diffuse hyperplasia, nodular goiter and multinodular goiter on scintigraphy. Therefore, scintigraphy, ultrasonography and serum thyroid hormone estimation alone may not be helpful for the final diagnosis of Hashimoto's disease. To eliminate unnecessary surgical intervention, all patients should be evaluated by means of physical examination and thyroid autoantibodies, in addition to a thyroid scan, ultrasonography, serum thyroid hormones and fine needle aspiration biopsy when necessary. PMID- 9392798 TI - An evaluation of FDG-PET in the detection and differentiation of thyroid tumours. AB - We evaluated the usefulness of FDG-PET for the detection of thyroid tumours and the differentiation between benign and malignant tumours. The subjects consisted of 5 normal volunteers and 22 patients, including 3 with follicular adenoma, 16 with papillary carcinoma and 3 with follicular carcinoma. The results were then evaluated both visually and semi-quantitatively using the standardized uptake value (SUV). All 22 tumours were seen as areas of high FDG uptake. FDG uptake in the normal thyroid gland, follicular adenoma, papillary carcinoma and follicular carcinoma was 1.0 +/- 0.2, 2.1 +/- 0.4, 4.7 +/- 3.2 and 4.6 +/- 2.9, respectively. Significant differences were observed between papillary carcinoma and both follicular adenoma (P < 0.05) and the normal thyroid gland (P < 0.001), and between follicular adenoma and the normal thyroid gland (P < 0.001). For the diagnosis of carcinoma, 58% sensitivity, 100% specificity and 73% accuracy were obtained when the highest FDG uptake value in adenoma was taken as the threshold. Our results thus indicate that high FDG uptake in a thyroid tumour suggests malignancy even though low levels of FDG uptake cannot completely rule out malignancy. PMID- 9392799 TI - Colonic transit time assessed by 67 Ga-citrate in a case-series of patients with recto-sigmoid adenomas. AB - One plausible mechanism by which dietary factors may influence colorectal carcinogenesis is through their effect on intestinal transit time. This study examined colonic transit by means of oral 67Ga-citrate in a case-series of patients who had developed recto-sigmoid adenoma. Adenoma patients had a significantly shorter transit time than constipated patients (P = 0.01) and our results also suggest (but do not show conclusively) that colonic transit in adenoma patients is similar to that of normal controls. Although these findings require confirmation from a larger study, they raise the hypothesis that colonic transit times are not delayed in patients who harbour recto-sigmoid adenomas. PMID- 9392800 TI - Detection of metastatic bone lesions in patients with prostate carcinoma: 99Tcm monoclonal antibody imaging. AB - Bone scintigraphy has been shown to be sensitive in determining bone involvement in patients with malignancy, but it does not allow the assessment of bone marrow lesions in early disease. The aim of this study was to detect bone marrow invasion using 99Tcm-labelled monoclonal antigranulocyte antibody (AgMoAb) in patients with prostate carcinoma. We studied 56 patients whose mean (+/- S.D.) age was 67 +/- 7 years. The mean prostate-specific antigen level was 6.1 ng ml-1 (normal range 0-5 ng ml-1). Twelve patients were in stage A, 16 in stage B, 17 in stage C and 11 in stage D. Six patients had been receiving chemotherapy and four patients radiotherapy before scanning. Bone scans were obtained 2 h after the intravenous injection of 555 MBq 99Tcm-methylene diphosphonate (99Tcm-MDP). Within a week, bone marrow imaging was performed 4 and 24 h after the injection of 555 MBq 99Tcm-AgMoAb. Metastatic bone lesions were detected on the 99Tcm-MDP scans of 14/56 (25%) patients, of whom one was in stage A, two in stage B, four in stage C and seven in stage D. Hypoactive lesions in bone marrow were detected in 25/56 (45%) patients, of whom two were in stage A, five in stage B, seven in stage C and 11 in stage D. Bone marrow metastases were confirmed in six patients by computed tomography (CT) and magnetic resonance imaging (MRI) and in two patients by marrow aspiration biopsy. A false-positive immune scintigram was found in three patients previously receiving radiotherapy or chemotherapy. We suggest that 99Tcm-AgMoAb scintigraphy is a sensitive procedure for the detection of bone marrow lesions. However, the reason for false-positive and false-negative results should be considered and CT, MRI and marrow biopsy should be performed when clinically necessary. PMID- 9392801 TI - Validation of a technique for the preparation of individual patient doses of 14 C urea. PMID- 9392802 TI - Permanent multisite cardiac pacing. PMID- 9392803 TI - Analysis of the intraventricular electrogram for differentiation of distinct monomorphic ventricular arrhythmias. AB - This study investigated the effectiveness of correlation waveform analysis for identifying different ventricular electrogram morphologies of multiple VTs in the same patient. Patients with implantable antitachycardia devices are commonly subject to the occurrence of more than one distinct monomorphic VT. Each of these VTs may have unique therapeutic alternatives for termination. VTs with identical and different monomorphic configurations were recorded (1-500 Hz) using distal bipolar (1 cm) and distal unipolar electrograms from the right ventricular apex. Thirty-six distinct monomorphic VTs induced in 15 patients were analyzed. Nine VTs with identical morphologies (12/12 surface ECGs) were induced twice and used as a control. A template was created for each VT induced. Correlation waveform analysis was used to compare each depolarization of all other VTs induced subsequently in the same patient. The mean correlation coefficient (p mu) of cycle-by-cycle analysis was used as a discriminant function: p mu > or = 0.95 was considered matched; and p mu < 0.95 was considered distinct. From the control population, VTs were successfully classified as identical in 9 of 9 cases (100%) using both bipolar and unipolar electrograms. VTs with different monomorphic configurations were successfully classified as being different in 31 of 33 cases (94%) using bipolar electrogram analysis and in 29 of 33 cases (88%) using the unipolar. Template matching is effective for detecting: (1) the recurrence of VTs, which are identical; and (2) the occurrence of a VT with a different configuration. This method appears effective using either unipolar or bipolar intracardiac waveforms. PMID- 9392804 TI - Atrial septal pacing: a method for pacing both atria simultaneously. AB - By pacing both atria simultaneously, one could reliably predict and optimize left sided AV timing without concern for IACT. With synchronous depolarization of the atria, reentrant arrhythmias might be suppressed. We studied four male patients (73 +/- 3 years) with paroxysmal atrial fibrillation and symptomatic bradyarrhythmias using TEE and fluoroscopy as guides; a standard active fixation screw-in lead (Medtronic model #4058) was attached to the interatrial septum and a standard tined lead was placed in the ventricle. The generators were Medtronic model 7960. The baseline ECG was compared to the paced ECG and the conduction time were measured to the high right atrium, distal coronary sinus and atrial septum in normal sinus rhythm, atrial septal pacing, and AAT pacing. On the surface ECG, no acceleration or delay in AV conduction was noted during AAI pacing from the interatrial septum as compared with normal sinus rhythm. The mean interatrial conduction time for all 4 patients was 106 +/- 2 ms; the interatrial conduction time measured during AAT pacing utilizing the atrial septal pacing lead was 97 +/- 4 ms (P = NS). During atrial septal pacing, the mean conduction time to the high right atrium was 53 +/- 2 ms. The mean conduction time to the lateral left atrium during atrial septal pacing, was likewise 53 +/- 2 ms. We conclude that it is possible to pace both atria simultaneously from a single site using a standard active fixation lead guided by TEE and fluoroscopy. Such a pacing system allows accurate timing of the left-sided AV delay. PMID- 9392805 TI - Asystolic cardiac arrest during head-up tilt test: incidence and therapeutic implications. AB - Occasionally, the cardioinhibitory response may be profound during tilt induced syncope. Whether this response is associated with more severe symptoms or predicts a poor response to pharmacotherapy remains controversial. The aim of this study was to characterize patients with vasovagally mediated asystole occurring during head-up tilt test and to evaluate the respective interests of sequential pacing and beta-blockers to treat them. We performed 60 degree tilt testing in 179 consecutive patients with unexplained syncope (91 women and 88 men, age 36.6 +/- 20.1 years). Asystole was defined as a ventricular pause > 5 seconds. All patients with tilt induced asystole received therapy with either beta-blockers or sequential pacing, the efficacy of which was evaluated with serial tilt tests. Of 77 patients with positive tilt test, 10 developed syncope related to asystole (mean duration 11.9 +/- 4.9 s), 2 with spontaneous recovery, and 8 with seizures needing a brief cardiopulmonary resuscitation. When compared with patients without asystole, asystolic patients had more severe symptoms (seizures: 6/10 vs 9/67, P = 0.05, injury 9/10 vs 27/67, P = 0.0048). In the first six patients in whom cardiac pacing was considered, syncope or presyncope still occurred despite atrioventricular pacing at 45 beats/min. Five of these 6 patients, as well as the remaining 4 asystolic patients, were tilted with beta blockers: 3 patients became tilt-negative; 3 were significantly improved; and 3 did not respond. During follow-up (mean 22.7 +/- 11.7 months) with every patient taking beta-blockers and seven having a permanent pacemaker, no syncopal recurrence was observed. Tilt-induced asystole that may require resuscitative maneuvers occurs especially in patients with a history of seizures or injury. Therapy with beta-blockers in often effective to prevent induction of syncope as well as recurrences. PMID- 9392806 TI - Permanent left atrial pacing with a specifically designed coronary sinus lead. AB - This article reports the original use of a specifically designed coronary sinus (CS) lead for permanent left atrial (LA) pacing. The device is characterized by its distal end shape featuring a double 45 degree angulation, which ensures very close contact with the CS upper wall. The device was successfully implanted in 39 out of 40 patients (97.5%). The tip electrode was eventually positioned in the distal CS in 9 patients, in the middle CS in 21 patients, and close to the ostium in the proximal CS in 9 patients. The mean acute pacing threshold voltage was 0.9 +/- 0.5 V with a mean impedance of 578 +/- 144 omega as measured in unipolar distal configuration at 0.5 ms pulse width (PW). The mean A wave amplitude was 3.5 +/- 2.1 mV. Early lead dislodgment occurred only once (3%) when the tip electrode was placed in the distal or middle CS, but more often (4/9 cases) when it was placed in the proximal CS. After a mean follow-up duration of 14 +/- 8.5 months, 35 of the 39 successfully implanted leads (89.7%) were still functional in terms of LA pacing and sensing. The mean chronic pacing threshold voltage was 1.5 +/- 0.8 V and the mean A wave amplitude was 2.7 +/- 1.6 mV. There were no lead related complications. In conclusion, the device proved to be safe and highly effective for permanent LA pacing, provided the distal tip could be positioned in the distal or middle part of the CS. PMID- 9392807 TI - Efficacy and safety of a new protocol for continuous infusion of midazolam and fentanyl and its effects on patient distress during electrophysiological studies. AB - Electrophysiological studies are often distressing for patients. We devised a regime of continuous infusion of midazolam and fentanyl during electrophysiological studies without the presence of a specialist anaesthetist. The effects on key hemodynamic and respiratory variables and level of sedation were evaluated in detail in the first 775 patients. The safety of this practice was evaluated in 1,344 consecutive patients. Doses were calculated according to patients' weight and age. A mean total dose of 26 mg of midazolam and 115 mcg of fentanyl were infused. Satisfactory sedation was achieved in 97% of patients. The mean duration of procedure was 188 +/- 90 minutes. Complete amnesia of the procedure was obtained in 87% of patients. Sedation caused clinically insignificant changes in respiratory rate, oxygen saturation, end-tidal CO2 and blood pressure. There were no major complications related to sedation. Upper airway obstruction, usually minor, occurred in 42% and some restlessness in 20% of sedated patients. The assistance of a specialist anesthetist was required in 0.3% of sedated patients for management of restlessness, hypoventilation, or obstructive sleep apnea. The amount of distress experienced by sedated patients (n = 775) was significantly less compared to a previous series of nonsedated patients (n = 775) undergoing electrophysiological studies (P < 0.001). The degree of distress experienced by patients during electrophysiological studies can be reduced significantly by sedation with intravenous midazolam and fentanyl. Continuous infusion is an efficient, safe, and effective way of administering midazolam and fentanyl. PMID- 9392808 TI - The combined transvenous implantation of cardioverter defibrillators and permanent pacemakers. AB - We developed criteria for implantation and programming of permanent endocardial pacemakers in patients with a nonthoracotomy ICD system. These criteria were prospectively used in 10 patients who recieved an ICD prior to (n = 5) or following (n = 5) implantation of a dual chamber (n = 6) or ventricular (n = 4) pacemaker with a unipolar (n = 4) or bipolar (n = 6) lead configuration. All patients were tested for interactions or malfunctions. Undersensing of ventricular fibrillation by the atrial sense amplifier and inadequate atrial pacing occurred in one patient with a unipolar dual chamber system programmed to AAIR but didn't impair ICD sensing. Transient or permanent loss of capture or sensing of the pacemaker was not observed after ICD shocks with the output programmed to double pulse width and voltage of stimulation threshold and the sensitivity to 50% of the detected R wave. One episode of transient reprogramming occurred without clinical consequences. One unipolar ventricular pacemaker lead had to be exchanged against a bipolar lead because of oversensing of the pacing artifact by the ICD. There was no failure of an ICD to detect ventricular arrhythmias due to inadequate pacemaker activity. During a follow-up period of 21 +/- 11 months, a total of 78 ventricular arrhythmias were effectively treated in six patients. Thus, a combined use of transvenous ICD and pacemaker is possible despite the close vicinity of pacing and defibrillations leads. Optimized programming different to the common settings is required. As interactions occurred only in unipolar pacemaker leads bipolar systems should be used in these patients. PMID- 9392809 TI - Neurohumoral and hemodynamic mechanisms of diuresis during atrioventricular nodal reentrant tachycardia. AB - Thirty-two consecutive patients with paroxysmal supraventricular tachycardias, with previously defined mechanisms of the tachycardias, were interviewed by noninvestigators about whether they experienced symptoms of diuresis during or at the termination of the tachycardias, to test the hypothesis that patients with AV nodal reentrant tachycardia would have a feeling of diuresis, polyuria, or both during or at the termination of the tachycardia. Twelve of the 13 patients with AV nodal reentrant tachycardia (92%), two of the 15 patients with AV reentrant tachycardia (13%), and one of the 4 patients with atrial flutter associated with 2:1 AV conduction (25%) felt diuresis during or at the termination of the tachycardias (AV nodal reentrant tachycardia vs other forms of tachycardia; P < 0.001). In 14 of the 32 patients, the right atrial pressure and plasma atrial natriuretic peptide (ANP) concentration were measured during both the tachycardias and sinus rhythm. The mean right atrial pressure during AV nodal reentrant tachycardia was significantly elevated compared to that during other forms of tachycardia (P < 0.01). The plasma ANP concentration during AV nodal reentrant tachycardia was also elevated significantly compared to that during other forms of tachycardias (P < 0.001). There were no significant differences in the cycle lengths of the tachycardias, age, left atrial dimensions, or the left ventricular ejection fraction between the AV nodal reentrant tachycardia and the other forms of tachycardia. We concluded that the feeling of diuresis during or at the termination of tachycardia was a more common symptom in patients with AV nodal reentrant tachycardia. The higher secretion of plasma ANP from the right atrium might be involved in the mechanism of this symptom. PMID- 9392810 TI - Procainamide induced change of the width of the zone of entrainment and its relation to the inducibility of reentrant ventricular tachycardia. AB - Procainamide depresses conduction velocity and prolongs refractoriness in myocardium responsible for reentrant VT, but the mechanism by which the induction of VT is suppressed after procainamide administration remains to be determined. In the present study, the relationship between electrophysiological parameters and the noninducibility of VT was assessed during procainamide therapy with a special reference to the change of an excitable gap. Clinically documented monomorphic sustained VT was induced in 30 patients and, utilizing the phenomenon of transient entrainment, the zone of entrainment was measured as the difference between the cycle length of VT and the longest paced cycle length interrupting VT (block cycle length) which was determined as the paced cycle length decreased in steps of 10 ms, and used as an index of the excitable gap. The effective refractory period was measured at the pacing site and the paced QRS duration was used as an index of the global conduction time in the ventricle. The cycle length of VT, the block cycle length, and the width of the zone of entrainment were determined and compared between the responders and nonresponders. In 15 patients, these parameters were determined at the intermediate dose and related to subsequent noninducibility at the final dose. At the final doses of procainamide, VT was suppressed in 8 (26.7%) of 30 patients. However, the cycle length of VT, the block cycle length, and the width of the zone of entrainment were unable to predict the drug efficacy, i.e., noninducibility. The change in the effective refractory period at the pacing site or the width of the paced QRS duration was not different between the responders and nonresponders. Among the variables, only the width of the zone of entrainment showed a significant narrowing in the responders at the intermediate dose of procainamide, and it was smaller than that of the nonresponders. The significant narrowing of the width of the zone of entrainment was associated with the subsequent noninducibility of VT at the final dose. The present study showed that the baseline cycle length of VT, the block cycle length, the drug induced change of the effective refractory period, or the paced QRS duration was not a predictor of the noninducibility after procainamide administration. However, a significant narrowing of the width of the zone of entrainment at the intermediate dose was associated with the noninducibility of VT at the final dose. PMID- 9392811 TI - Clinical performance of steroid-eluting pacing leads with 1.2-mm2 electrodes. AB - To raise pacing impedance and reduce battery current drain, new tined steroid eluting leads were developed with 1.2-mm2 hemispherical electrodes, instead of conventional 5-8 mm2. Twenty-two unipolar J-shaped atrial leads and 25 unipolar ventricular leads (models 4533 and 4033, respectively) were implanted in 33 consecutive patients and followed for a mean of 25 months (range 18-29). Handling characteristics of atrial leads were found favorable. The leads slipped easily into the right atrial appendage and were easy to position. Handling characteristics of ventricular leads were satisfying, but more efforts had to be applied to cross the tricuspid valve. Special care was taken to avoid perforation of the myocardium due to the small lead tip. Following implantation, four ventricular and one atrial lead exhibited instability of pacing thresholds that resolved spontaneously within 1-3 days of implantation. Except for this, no lead malfunctioned. The reoperation rate was zero. The mean electrogram amplitudes of 15 mV (ventricle) and 4 mV (atrium), and the mean chronic pacing threshold of 0.085 ms at 1.6 V (app. 0.43 V at 0.5 ms) were comparable with the best values seen in the literature on passive fixation leads. The rest of the electrophysiological parameters were enhanced: mean pacing impedances were 984 omega (acute) and 900 Q (chronic), mean slew rates 3.26 V/s (ventricle) and 1.75 V/s (atrium), mean acute voltage threshold at 0.5 ms was 0.25 V, mean current and energy thresholds calculated at 0.5 ms were 260 microA and 32 nJ (acute) and 478 microA and 103 nJ (chronic). The electrical characteristics of these leads provide for increased pacemaker longevity in combination with substantial safety margins for pacing and sensing. PMID- 9392812 TI - Long QTc and torsades de pointes in human immunodeficiency virus disease. AB - Three patients with human immunodeficiency virus (HIV) infection presented with QT, prolongation (> 440 ms) and torsades de pointes. We sought to evaluate the etiology of the long QT syndrome in these patients without previously identified causes for QT, prolongation, and determine the prevalence among patients with HIV infection. The three index patients underwent: (1) left stellate ganglion block; (2) beta-blocker challenge; and (3) electrocardiographic stress testing. QTc interval was measured before and after intervention. We undertook a retrospective analysis of prevalence of QTC prolongation among all patients with computerized ECGs over a 6-month period at one institution and compared it to the prevalence in hospitalized patients with HIV disease. Thirty-four thousand one hundred eighty-one patients with computerized ECGs were screened for QTc prolongation. Forty-two hospitalized patients with HI disease had computerized ECG during the same 6-month period. In the three index patients, the QTc failed to shorten with left stellate ganglion blockade, beta-blocker challenge, or stress testing, suggesting an acquired form of the long QT syndrome in these patients with HIV disease. None had previously recognized acquired causes of QT, prolongation. Mexiletine hydrochloride was useful in preventing recurrences of torsades de pointes. We observed a 7.0% prevalence of QT, prolongation among all patients screened. Hospitalized patients with HIV disease (n = 42) during this same period, demonstrated an increased prevalence of QT, prolongation (28.6%, P = 0.002). Patients with HIV disease have a significantly higher prevalence of QTc prolongation than a general hospital-based population, may have an unrecognized acquired form of the long QT syndrome, and are at risk for torsades de pointes. PMID- 9392813 TI - Retroconduction selective recognition in wide-dipole floating atrial sensing. The Multicenter Study Group. AB - Effective discrimination of retrogradely conducted P waves would allow distinguishing sinus tachycardia from supraventricular tachycardias due to AV or nodal reentry, and would prevent pacemaker-mediated tachycardia in AV sequential pacing. This might be especially relevant in VDD implants, where retroconduction could be induced by escape ventricular stimulation. In order to analyze the respective waveform properties, anterograde and retrograde atrial signals were recorded by a wide floating electrode dipole, on the implantation of a permanent single-pass lead for VDD pacing. Generally, bipolar recording did not allow reliable discrimination, while the signal nature could be readily diagnosed from the main features of the unipolar atrial electrograms. The unipolar waveform recorded under sinus rhythm in high right atrium, close to the superior vena cava opening (proximal EGM), started with a negative deflection in 88% of the patients. In 7% of the patients, the first deflection of the signal was positive in some cardiac cycles only, and, on the average, the amplitude of the positive phase was not higher than 5% of the signal peak-to-peak amplitude. Conversely, under retroconduction, the starting deflection attained higher positive values in 98% of the patients, being stably over 15% of the peak-to-peak amplitude in 86% of the Furthermore, in 69% of the cases, the lag time between the onset of the negative deflection of proximal and distal (mid-low atrium) unipolar EGM changed unambiguously when retroconduction occurred, exceeding the range of variation observed in each patient during sinus activity. The combined evaluation of unipolar EGM shape and lag time allowed specific retroconduction recognition in 95% of the patients. We suggest that this approach may yield useful information for the discrimination of retrograde atrial signals, provided that the recording dipole is sufficiently long and the proximal electrode is properly positioned in the high right atrium. PMID- 9392814 TI - The top ten fallacies of nonsustained ventricular tachycardia. AB - Nonsustained ventricular tachycardia (NSVT) continues to remain a subject of controversy. This is true despite a wealth of epidemiologic and basic/clinical laboratory findings that have accumulated during the past 2 decades. However, these data not only generate the impetus to conduct further research, but also provide compelling arguments against continued adherence to time honored precepts about NSVT that evolved since the inception of the "PVC Hypothesis," although never substantiated by rigorous scientific inquiry. This paper discusses the "top ten" fallacies of NSVT and details the data that support abandonment of them. PMID- 9392815 TI - Graphical representation of complex data--diurnal patterns of initiations of atrial fibrillation episodes. AB - A construction of a purpose designed graphical display is demonstrated in a study investigating the circadian distribution of patterns of RR interval sequences preceding episodes of paroxysmal atrial fibrillation (PAF). Based on a comparison with a (80%, 120%) range around the median of preceding 10 RR intervals, each RR interval is classified as normal, short, or long. Classifications of RR intervals in n-tuplets (n = 1, ...,5) preceding PAF episodes are used to compute probabilities of individual types of sequences occurring within 4-hour periods of the day (between 1 am, 5 am, 9 am, 1 pm, 5 pm, and 9 pm). Graphical representation of the data is proposed using a hierarchy of bar graphs. The graphical system has been filled with data of 327 atrial fibrillation episodes recorded in 46 24-hour ECGs in PAF patients. The graphical analysis supports a link between PAF initiation and cardiac autonomic status. PMID- 9392816 TI - Practice expense: applying accounting principles in a specialty practice. PMID- 9392817 TI - Idiopathic left ventricular tachycardia: what is the mechanism? PMID- 9392818 TI - Interaction between electronic article surveillance systems and implantable defibrillators: insights from a fourth generation ICD. AB - We present a case report of an inappropriate discharge from a fourth generation implantable cardioverter defibrillator (ICD) as a result of exposure to electromagnetic interference. A 60-year-old man implanted with a Medtronic 7219D defibrillator experienced shocks without preceding symptoms while walking through an electronic surveillance system in a store. Though this has been reported, the mechanism of the interaction remains unexplained. The electrogram storage capabilities of this particular device enabled us to establish that this resulted from inappropriate sensing of the electromagnetic interference. PMID- 9392819 TI - Ventricular output failure in a DDD permanent pacemaker associated with increased atrial output. AB - Previous reports have described the occurrence of ventricular output failure in a permanent DDD pacemaker system related to an increase in the atrial output in the presence of low atrial lead impedance (Medtronic Synergyst/Synergyst II). This phenomenon is seen exclusively following atrial paced events and may potentially lead to significant bradyarrhythmia or ventricular asystole in a pacemaker dependent patient. We describe the occurrence of analogous behavior in a Medtronic Symbios 7006 generator. PMID- 9392820 TI - A pitfall in using far-field bipolar electrograms in arrhythmia discrimination in a patient with an implantable cardioverter defibrillator. AB - Analysis of stored electrograms from bipolar far-field electrodes is considered to be useful in differentiating supraventricular from ventricular arrhythmias. A case of inappropriate ICD shocks for sinus tachycardia is presented whereby successive shocks caused marked widening of the ventricular electrograms. Analysis of these stored electrograms, recorded from far-field bipolar electrodes, gave the false impression of ventricular tachycardia. The widening was due to the current of injury effects, probably a consequence of the large amount of intervening myocardium between the bipoles. While analyzing recordings from far-field bipolar electrodes is generally useful, it is not always reliable, for changes in electrogram morphology, relative to baseline rhythm, may result from other factors like current of injury. PMID- 9392821 TI - Near doubling of heart rate after intravenous verapamil for treatment of atrial fibrillation without preexcitation. AB - Initial treatment of atrial fibrillation often involves pharmacological therapy to control ventricular response. While verapamil is usually safe and effective when used for this purpose, we report a proarrhythmic response. In this report a 30-year-old female presented with palpitations associated with atrial fibrillation and a ventricular response of 145 beats/min. Soon after she was given 5 mg of intravenous verapamil her ECG documented a regular wide QRS tachycardia at 290 beats/min. After 7 seconds the rhythm returned to an irregularly irregular narrow QRS tachycardia at 125-150 beats/min. At a later electrophysiology study there was neither evidence of preexcitation nor inducible supraventricular or ventricular tachycardia. These data suggest that verapamil may have been associated with acceleration of the heart rate. The mechanism of proarrhythmia may be related to an alteration in the atrial rhythm from atrial fibrillation to atrial flutter, with additional factors as well. PMID- 9392822 TI - Runaway in a modern "soft top" pacemaker. AB - Although runaway pacemaker was a relatively common occurrence, modern pacemaker design has made it extremely rare. We report a case occurring with a modern "soft top" pacemaker that resulted from a loss of hermeticity. Although the different treatment options are discussed, the definitive maneuvre is explantation of the faulty pacemakers PMID- 9392824 TI - Classification of atrial fibrillation. PMID- 9392823 TI - Pectoral muscle stimulation after falling off a ladder. AB - A-pacemaker that had been implanted correctly was found at follow-up to have "flipped over" as a result of a fall from a ladder. Symptomatic pectoral muscle stimulation ceased immediately when the generator was manually flipped back into the correct position. PMID- 9392825 TI - STIMAREC report. PMID- 9392826 TI - DIFIMAREC report. PMID- 9392827 TI - Synthetic peptide from the V3 loop consensus motif with a potent anti-HIV activity inhibits ristocetin-mediated vWF-GPIb interaction. AB - The V3 loop consensus motif. Arg-Gly-Pro-Gly-Arg-Ala-Phe-Val-Thr-Ile (HIV-1 IIIB), inhibits an interaction of HIV with CD4-positive lymphocytes. Recently, both proline-rich peptides and peptides containing proline-glycine loops (beta turns) form a complex with ristocetin dimers. These peptides interact with ristocetin-loaded platelet membrane glycoprotein (GP) Ib and act as inhibitors of von Willebrand factor (vWF)-GPIb interaction by preventing the subsequent formation of ristocetin dimer bridges. The Pro-Gly sequence is also present in the V3 loop consensus motif, Arg-Gly-Pro-Gly-Arg-Ala-Phe-Val-Thr-Ile (HIV-1 IIIB). In this report, we have evaluated the effect of the HIV-1 IIIB peptide on vWF binding to GPIb. This peptide only inhibited vWF binding to GPIb as well as platelet aggregation in the presence of ristocetin while it had no effect on botrocetin-mediated vWF interaction with platelets. The peptide inhibited a binding of anti-vWF monoclonal antibody (RG-46) to immobilized vWF. Furthermore, ristocetin inhibited the binding of HIV-1 IIIB peptide to immobilized CXC chemokine receptor-4 (CXCR-4) peptide. These results indicate that ristocetin may prevent HIV infection and would be useful a tool to understand the mechanism of HIV tissue tropism and infection. PMID- 9392828 TI - Expression of rat pro cholecystokinin (CCK) in bacteria and in insect cells infected with recombinant baculovirus. AB - Neuropeptide prohormones are generally not abundant in nature as they exits to be processed and contain sites which could be cleaved by a number of cellular proteases. In order to study the processing of prohormones in vitro it is necessary to produce them in quantity. Finding an expression system which produces intact prohormone has been a matter of trial and error. We report that intact rat pro CCK was produced with an amino-terminal His-Tag in e. coli, and it was secreted from sf9 and other insect cells infected with a recombinant baculovirus vector. The bacteria contained about 0.1 micrograms pro CCK/ml of cells. The High 5 insect cells produced 4.3 micrograms/ml medium (as determined by RIA), 10 times as much as sf9 or sf21. Using a combination of ion exchange, gel filtration and HPLC, the insect cell protein was purified about 150 fold with a recovery of about 16%. The secreted insect cell pro CCK is tyrosine sulfated like its mammalian equivalent. Using these expression systems it is possible to produce significant (microgram to mg) quantities of pro CCK for immunologic, enzymatic and structural studies. PMID- 9392829 TI - Lepidopteran peptides of the allatostatin superfamily. AB - Peptides of the allatostatin superfamily with the C-terminal amino acid sequence YXFGL-NH2 have been isolated and identified from the lepidopterans, the codling moth, Cydia pomonella (Tortricidae) and the bollworm, Helicoverpa armigera (Noctuidae). The peptides, designated cydiastatins and helicostatins respectively, were monitored during purification with radioimmunoassays based on the callatostatins of the blowfly Calliphora vomitoria. The eight peptides from each of the two species appear to form an homologous series with four identical and three that differ by a single amino acid. This study demonstrates the ubiquitous nature of this family of peptides in insects. PMID- 9392830 TI - Protein phosphorylation in snail cardiocytes stimulated with molluscan peptide SCPb. AB - Dissociated muscle cells prepared from hearts of the pulmonate snail Helix aspersa were used to study signal transduction induced by molluscan cardioactive peptides. The effects of SCPb on the cAMP levels of whole hearts and the cell preparation were compared. The cells responded to SCPb with a dose-dependent increase in cAMP that had the same structure-activity relations as seen in the intact tissue. SCPb increased the phosphorylation of a 53 kDa protein in a dose dependent manner; threshold was 10(-9) M. The SCPb-induced phosphorylation was mimicked by forskolin and 8-CPT-cAMP. FMRFamide stimulation had no effect on the phosphorylation of this protein. PMID- 9392831 TI - Thyrotropin-releasing activity of histone H2A, H2B and peptide MB35. AB - Histones possess multiple hormone-like activities. We studied the specificity and signal transduction pathways involved in the thyrotrophin (TSH)-releasing activity of histones H2A, H2B and peptide MB35, a H2A fragment, using perifused and incubated dispersed rat pituitary cells and measuring TSH release by a specific R1A. Histones released TSH in a dose- and time-dependent fashion while peptide MB35 behaved as a weaker secretagogue. Trifluoperazine and EGTA blocked histone-stimulated TSH release while forskolin and other cAMP enhancers did not. We conclude that the TSH-releasing activity of histones H2A and H2B is mediated by calcium- and diacylglycerol-associated pathways. PMID- 9392832 TI - Influence of [4Cl-D-Phe6,Leu17]VIP on VIP- and central TRH-induced gastric hyperemia. AB - The specific VIP receptor antagonist, [4Cl-D-Phe6,Leu17]VIP, infused i.v. blocked close-intra-arterial infusion of VIP-induced increase in gastric mucosal blood flow (GMBF, measured by the hydrogen gas clearance), and decrease in mean arterial blood pressure while not influencing basal levels in urethane anesthetized rats. The thyrotropin-releasing hormone (TRH) stable analog, RX 77368, injected intracisternally (IC, 30 ng) increased GMBF and blood pressure. The VIP antagonist did not significantly reduce the GMBF response to IC RX 77368 while enhancing the rise in blood pressure. These findings indicate that [4Cl-D Phe6,Leu17]VIP is an antagonist for exogenous VIP-induced gastric hyperemia and hypotension and that VIP modulates the systemic blood pressure response to IC RX 77368 at 30 ng while not playing a primary role in the increase of GMBF. PMID- 9392833 TI - Effects of non-glycated and glycated glucagon-like peptide-1(7-36) amide on glucose metabolism in isolated mouse abdominal muscle. AB - This study investigated the actions of non-glycated and glycated glucagon-like peptide-1(7-36)amide (tGLP-1) on glucose uptake and metabolism in isolated mouse abdominal muscle. Monoglycated tGLP-1 (Mr 3463.8) was prepared under hyperglycemic reducing conditions and purified by HPLC. Non-glycated tGLP-1 (10( 10)-10(-8) mol/l) stimulated both 2-deoxy-D-[1-3H]glucose uptake (1.3-1.5 fold) and 14C-glucose oxidation (1.4-1.7 fold) in muscle compared to controls without tGLP-1. Glycation reduced these stimulatory effects by 27-33% and 25% (at 10(-9) mol/l), respectively. tGLP-1 (10(-10)-10(-8) mol/l) promoted muscle glycogenesis and lactate production, whereas glycated peptide was ineffective below 10(-9) mol/l. This study demonstrates that tGLP-1 has potent glycogenic effects in mouse abdominal muscle in vitro and that glycation impairs its action. PMID- 9392834 TI - Elevated neuropeptide Y gene expression and release during hypoglycemic stress. AB - Our previous studies show that neuropeptide Y (NPY) is involved in mediating sympathetic nerve stimulation-induced vasoconstriction. Insulin hypoglycemia is known to produce increased sympathetic output and elevated arterial pressure. The present study examined the role of NPY in the hypertensive response to insulin by examining the effects of insulin on NPY gene expression, tissue content and release. Subcutaneous injection of insulin produced an immediate increase in plasma NPY immunoreactivity (NPYir) and delayed increases in adrenal and neuronal NPY mRNA and adrenal NPYir in rats. These results suggest that NPY may play a role in insulin-induced hypertension. PMID- 9392835 TI - Enterostatin actions in the amygdala and PVN to suppress feeding in the rat. AB - The pentapeptide enterostatin (ENT) inhibits feeding after injection into the cerebral ventricles. To localize the central sites of action of ENT, the peptide (0.01 to 3.3 nM) was microinjected into several brain regions and the intake of a high fat diet was measured. The results show that ENT injection in the paraventricular nucleus (PVN) or the amygdala (AMYG) produced a bi-phasic dose related feeding response, low doses of ENT inhibited feeding while higher dose had no effect. The effective dose to inhibit feeding in the AMYG was 10 fold lower than that in the PVN. No changes in food intake were observed after ENT injection into the ventromedial hypothalamus and nucleus tractus solitarius. The data provide further support that there are targets in the CNS for ENT and suggest that central ENT function is site specific. PMID- 9392836 TI - Stimulation of tachykinin NK-3 receptors in the nucleus basalis magnocellularis reduces alcohol intake in rats. AB - Injections in the nucleus basalis magnocellularis (NBM) of the tachykinin (TK) NK 3 receptor agonist [Asp5,6,MePhe8]substance P(5-11), also referred to as amino senktide (NH2-SENK), markedly reduced alcohol intake in genetically selected alcohol-preferring rats, offered 10% ethanol 2 h/day. The threshold dose in the NBM was 0.5 ng/site, while neither 1 nor 10 ng/rat of NH2-SENK inhibited alcohol intake following administration into the lateral ventricle. Injection of NH2 SENK, 25 ng/site, in the NBM did not modify water or food intake in water deprived rats, providing evidence for the behavioral selectivity of the effect on ethanol intake. The selective TK NK-3 receptor antagonist, R-820, injected in the NBM at the dose of 1000 ng/site 5 min before NH2-SENK 5 ng/site, significantly reduced the effect of NH2-SENK. The selective TK NK-1 receptor agonist [Sar9,Met(O2)11]substance P inhibited alcohol intake following injection in the NBM only at 25 ng/site; but the same dose induced marked grooming and inhibited also water intake in water deprived rats. The present results confirm that TK NK 3, but not NK-1, receptor agonists selectively inhibit ethanol intake in alcohol preferring rats and suggest that the NBM is a site of action for their effect. PMID- 9392837 TI - Comparison of responses to T-kinin and bradykinin in the mesenteric vascular bed of the cat. AB - Responses to T-kinin and bradykinin were compared in the mesenteric vascular bed of the cat. Under constant-flow conditions, injection of T-kinin and bradykinin into the perfusion circuit induced similar dose-related decreases in perfusion pressure. Responses to T-kinin and bradykinin were inhibited by the kinin B2 receptor antagonist Hoe-140, but were not altered by the B1 receptor antagonist des-Arg9-[Leu8]-BK, the histamine H1 antagonist pyrilamine, the histamine H2 receptor antagonist cimetidine, or the H3 receptor antagonist thioperamide. Vasodilator responses to T-kinin and bradykinin were attenuated by the nitric oxide synthase inhibitor, N omega Nitro-L-arginine methyl ester (L-NAME), but were not altered by the cyclooxygenase inhibitor, sodium meclofenamate, or the K+ ATP channel antagonist, U37883A. These data suggest that vasodilator responses to T-kinin and bradykinin are mediated by kinin B2 receptor stimulated release of nitric oxide from the endothelium, but that the activation of kinin B1 receptors, the release of vasodilator prostaglandins, or the opening of K+ ATP channels are not involved in the response to T-kinin in the mesenteric vascular bed of the cat. PMID- 9392838 TI - Differential regulation of angiotensinogen and natriuretic peptide mRNAs in rat brain by osmotic stimulation: focus on anterior hypothalamus and supraoptic nucleus. AB - Central angiotensin II and natriuretic peptide systems have been shown to be involved in the central regulation of blood fluid homeostasis with alterations in central peptide and/or receptor levels observed following changes in osmotic status. The present study investigated the effects of sodium loading on mRNA encoding the angiotensin II precursor, angiotensinogen (AOGEN), and the natriuretic peptides, atrial natriuretic peptide (ANP) and C-type natriuretic peptide (CNP) in rat brain using quantitative in situ hybridization histochemistry of [35S]- and [33P]-labeled oligonucleotide probes. Following 7 and 14 days of 2% sodium chloride in drinking water a significant increase was detected in preproAOGEN (ppAOGEN) mRNA in presumed astrocytes in regions of the anterior hypothalamus, including the periventricular nucleus, the medial preoptic area and medial preoptic nucleus, while a decrease was observed in astrocytes in the supraoptic nucleus. Other forebrain regions examined including the subfornical organ, bed nucleus of the stria terminalis and the arcuate nucleus showed no significant alteration in the level of ppAOGEN mRNA. Sodium loading did not appreciably alter ppANP or ppCNP mRNA levels in neurons of the anteromedial preoptic or arcuate nuclei or hippocampus at the times studied. PpANP mRNA levels were also unaltered in Barrington's nucleus following sodium loading, while preprocorticotropin-releasing hormone mRNA was significantly decreased. These results indicate that AOGEN mRNA transcription/stability in vivo is modulated by alterations in osmotic balance, consistent with previous reports of a central role for AII in cardiovascular and body fluid homeostasis. In contrast, despite reports of modulation of hypothalamic ANP-immunoreactivity following changes in osmotic status, it would appear that osmotic stimulation over periods of 7-14 days does not markedly alter the transcription or stability of hypothalamic natriuretic peptide mRNAs in vivo. PMID- 9392840 TI - Arginine vasotocin gene expression in hypothalamic neurons is up-regulated in chickens drinking hypertonic saline: an in situ hybridization study. AB - Osmotic stress stimulates the release of the avian hypothalamic neuropeptide arginine vasotocin (AVT) into the peripheral circulation. We conducted the present study to investigate the effects of salt-loading on AVT secretion and AVT gene expression in specific hypothalamic nuclei in chickens. White Leghorn chickens were provided food ad lib and either water or 2% NaCl to drink. Both plasma osmolality and plasma AVT levels were significantly increased in chickens that drank 2% NaCl for either two or four days compared to that in chickens that drank water. Results from in situ hybridization analysis demonstrated an increase in the number of neurons expressing AVT mRNA in the supraoptic (SON) and paraventricular nuclei (PVN) in chickens provided 2% NaCl to drink compared to chickens that were provided water to drink. The number of grains per neuron increased in the PVN, but not in the SON of osmotically stimulated birds. Thus, increased osmolality resulting from ingestion of hypertonic saline is an effective stimulus to increase hypothalamic AVT mRNA content in chickens. PMID- 9392839 TI - Oxytocin releases atrial natriuretic peptide from rat atria in vitro that exerts negative inotropic and chronotropic action. AB - Our previous experiments suggested that natriuresis induced by blood volume expansion, was brought about by oxytocin (OT)-stimulated atrial natriuretic peptide (ANP) release from the right atrium. We hypothesized that the ANP released might exert effects on the atrium itself and therefore carried out in vitro experiments to test this hypothesis. Heart rate and isometric tension were recorded from isolated rat atria mounted in an organ bath. Oxytocin exerted a dose-related, negative chrono- and inotropic effect with a minimal effective concentration (MEC) of 3 microM, 10-fold higher than required for ANP to exert comparable effects. The effects of OT were not blocked by atropine suggesting that they were not mediated via release of acetylcholine. Eight-bromoguanosine 3' 5'-cyclic monophosphate (cGMP) had similar effects to those of OT and ANP, suggesting that the effects of ANP were mediated by cGMP. When isolated ventricles, left or right atria, were incubated in vitro, OT had a dose-related effect to stimulate the release of ANP into the medium only from right atria with a MEC of 0.1 microM. A specific OT antagonist, F792 (1 microM), inhibited basal release of ANP and blocked the stimulatory action of OT on ANP release. The results support the hypothesis that OT, acting on its putative receptors in the right atrium, stimulates the release of ANP which then exerts a negative chrono- and inotropic effect via activation of guanylyl cyclase and release of cGMP. The ability of the oxytocin antagonist to reduce basal release of ANP from atria incubated in vitro supports the hypothesis that these effects could be physiologically significant. We hypothesize that blood volume expansion via baroreceptor input to the brain causes the release of OT which circulates to the heart and stimulates the release of ANP from the right atrium. This ANP then has a negative ino- and chronotropic effect in the atrium and possibly a negative inotropic effect in the right ventricle, left atrium and left ventricle, to produce an acute reduction in cardiac output that, coupled with its peripheral vasodilating actions, causes a rapid reduction in effective circulating blood volume. The ANP released would also act on the kidneys to cause natriuresis and ANP acts within the brain to inhibit water and salt intake leading to a gradual recovery of circulating blood volume to normal. PMID- 9392841 TI - The block of central vasopressin V1 but not V2 receptors suppresses grooming behavior and hypothermia induced by intracerebroventricular vasopressin in male rats. AB - The role of central vasopressin V1 receptors in grooming behavior induced by vasopressin and oxytocin was studied in male rats of the Wistar strain. The intracerebroventricular (ICV) injection of vasopressin (3 micrograms/5 microliters) induced hypothermia and enhanced novelty-induced grooming behavior. Enhanced grooming but not hypothermia was also induced by ICV injection of oxytocin (3 micrograms/5 microliters). The central administration of a selective vasopressin V1 receptor antagonist prevented the stimulating action of vasopressin on novelty-induced grooming and its hypothermic effect. The ICV injection of a selective vasopressin V2 receptor antagonist failed to affect vasopressin-induced grooming and hypothermic effect. An increase in core temperature was observed in oxytocin-injected animals pretreated with the vasopressin V1 receptor antagonist. Furthermore, pretreatment with the antagonist did not affect grooming induced by oxytocin. These results suggest that enhancement of grooming behavior and influence on thermoregulation are differently regulated by central receptors for vasopressin and oxytocin. PMID- 9392842 TI - Endomorphin 1 and 2, endogenous ligands for the mu-opioid receptor, decrease cardiac output, and total peripheral resistance in the rat. AB - Endomorphin 1 and 2 are recently discovered endogenous ligands for the mu-opioid receptor. In the present study, responses to intravenous administration of endomorphin 1 and 2 were investigated in the systemic vascular bed of the rat. Endomorphin 1 and 2 induced dose-related decreases in systemic arterial pressure when injected in doses of 10-100 nmol/kg i.v.. The decreases in systemic arterial pressure in response to endomorphin 1 and 2 were associated with significant decreases in heart rate, cardiac output, and total peripheral resistance. The endogenous ligand for the ORL1 receptor, nociceptin/OFQ had similar effects on systemic arterial pressure, heart rate, cardiac output, and total peripheral resistance in the rat. Injections of isoproterenol (1 microgram/kg i.v.) and calcitonin gene-related peptide (CGRP; 0.3 nmol/kg i.v.), decreased systemic arterial pressure and total peripheral resistance. However these decreases in arterial pressure were associated with increases in heart rate and cardiac output. The results of the present study demonstrate that the endomorphin peptides have significant vasodilator activity in the systemic vascular bed of the rat and show that this response is associated with a decrease in heart rate and cardiac output. PMID- 9392843 TI - Beta-endorphin-containing proteins in the human pituitary. AB - Two new proopiomelanocortin (POMC)-derived beta-endorphin (BE)-containing proteins were detected in the human pituitary, using HPLC, trypsin digestion, and a high sensitivity search with liquid secondary ion mass spectrometry (LSIMS) for the protonated molecule ion, (M + H)+, of tryptic peptides that are unique to BE. Proteins were extracted from pituitary tissues and were purified by solid phase extraction (SPE) chromatography and RP-HPLC. Each HPLC fraction was treated with trypsin, and each unseparated peptide mixture was analyzed by LSIMS to detect the two selected marker peptides (BE 20-24 and BE 10-19) that have excellent LSIMS desorption-ionization properties. The detection of both of those peptides indicated the presence of BE-containing proteins in two HPLC fractions (number 47 and 51). Tandem MS determined the amino acid sequence of the marker peptide BE 20 24 (NAIIK), and those sequence data optimized the specificity of the method. The two new BE-containing proteins derive from the C-terminal region of POMC, and were minor components in the two HPLC fractions. The major component in fraction 51 derived from the vasopressin-neurophysin 2-copeptin precursor. PMID- 9392844 TI - Pro-opiomelanocortin (POMC) expression and immunolocalization of POMC-related peptides in the ovary of Protopterus annectens, an African lungfish. AB - Antisera against adrenocorticotropic hormone (ACTH), alpha-melanocyte-stimulating hormone (alpha MSH) and beta-endorphin were used to localize pro-opiomelanocortin (POMC)-derived peptides in the ovary of the African lungfish Protopterus annectens by immunohistochemistry. Immunoreactivity was observed in the granulosa and the internal theca of the vitellogenic follicles. No immunoreactivity was observed in immature follicles. Using human POMC cDNA as the hybridization probe POMC-like mRNA was identified in situ in cells of the granulosa and internal theca of the vitellogenic follicles. No labeling was observed in primordial follicles. The demonstration in the same cells of POMC mRNA and POMC-related peptides immunoreactivity indicates a local production of the opiate hormones. PMID- 9392845 TI - Interactions of islet hormones with acetylcholine in the isolated rat pancreas. AB - This study investigates the effects of the islet hormones, insulin (INS), glucagon (GLU) and somatostatin (SOM) on acetylcholine (ACh)-evoked amylase secretion and calcium (Ca2+) mobilization in the isolated rat pancreas. Stimulation of pancreatic segments and acini with either INS, GLU or SOM resulted in small increases of amylase output compared to much large increases in enzyme output with ACh. Combinations of the peptide hormones with ACh resulted in enhanced secretory responses compared to the effects obtained with either ACh or each of the islet hormone alone. Genistein, the tyrosine kinase inhibitor, evoked a decrease in amylase output from pancreatic segments. It had no effect on the ACh evoke secretory response but it markedly inhibited the potentiation of the islet hormones with ACh. In pancreatic acinar cells either INS, GLU or SOM elicited moderate increases in amylase output compared to much larger responses with ACh. Furthermore, the islet hormones failed to potentiate the secretory effect of ACh in pancreatic acini. In fura-2 AM loaded acinar cells both INS and GLU evoked small increases in intracellular free calcium concentration [Ca2+]i compared to a much larger elevation with ACh. Both INS and GLU enhanced the ACh evoked [Ca2+]i. Genistein elicited a decrease in [Ca2+]i both in the absence and presence of both INS and GLU. It also decreased the rise in [Ca2+]i resulting from the combined presence of ACh with both INS and GLU. SOM had no significant effect on the ACh-induced [Ca2+]i. When genistein was combined with ACh and SOM there was a decrease in [Ca2+]i compared to the response obtained with SOM and ACh alone. The results indicate that both tyrosine kinase and cellular Ca2+ seem to be the intracellular mediators associated with the enhanced secretory responses obtained with a combination of the islet hormones with ACh. Finally, our results using immunohistochemical techniques confirm the presence of INS-, GLU- SOM- and ACh-immunoreactive cells in the endocrine and neural elements of the rat pancreas. PMID- 9392846 TI - Transport of insulin across the blood-brain barrier: saturability at euglycemic doses of insulin. AB - Blood-borne insulin is known to cross the blood-brain barrier (BBB) where it can act as a satiety peptide. We examined in mice the pharmacokinetics and characteristics of such passage by multiple-time regression analysis. The unidirectional influx constant (Ki) of human insulin radioactively labeled with iodine (I-Ins) ranged from 0.87 to 1.7 microliters/g-min. The transport of I-Ins was inhibited almost 50% by 0.1 micrograms/mouse of unlabeled human insulin, a dose that had no effect on serum glucose. Similar results were found with rat insulin. The results with self-inhibition suggest that any hemoencephalic signal transmitted by the blood to brain transport of insulin is independent of the effects of insulin on glucose. The transport of I-Ins was altered by aluminum but not by administration of tyrosine, verapamil, or leptin, indicating independence from amino acid transport, the p-glycoprotein system, a slow calcium channel, or leptin transport. By contrast with insulin, enzyme degradation limited the uptake and accumulation by brain of intravenously injected, radioactively labeled glucagon and glucagon-like peptide. In conclusion, these results are consistent with the view that insulin can affect satiety and related behaviors independently of its peripheral effects by crossing the BBB to act within the brain. PMID- 9392847 TI - Bioavailability and transport of peptides and peptide drugs into the brain. AB - Rational drug design and the targeting of specific organs has become a reality in modern drug development, with the emergence of molecular biology and receptor chemistry as powerful tools for the pharmacologist. A greater understanding of peptide function as one of the major extracellular message systems has made neuropeptides an important target in neuropharmaceutical drug design. The major obstacle to targeting the brain with therapeutics is the presence of the blood brain barrier (BBB), which controls the concentration and entry of solutes into the central nervous system. Peptides are generally polar in nature, do not easily cross the blood-brain barrier by diffusion, and except for a small number do not have specific transport systems. Peptides can also undergo metabolic deactivation by peptidases of the blood, brain and the endothelial cells that comprise the BBB. In this review, we discuss a number of the recent strategies which have been used to promote peptide stability and peptide entry into the brain. In addition, we approach the subject of targeting specific transport systems that can be found on the brain endothelial cells, and describe the limitations of the methodologies that are currently used to study brain entry of neuropharmaceuticals. PMID- 9392848 TI - Inhibition of bacterial growth by synthetic SP-B1-78 peptides. AB - Residues 12-34 of mature human pulmonary surfactant protein B (SP-B1-78) are 68% homologous to residues 48-72 of the frog peptide antibiotic dermaseptin b I. We examined the effects of SP-B1-78 on the growth of Escherichia coli in order to find whether full length SP-B1-78 might act as a peptide antibiotic. We found that SP-B1-78 peptide inhibited growth of E. coli (MIC = 210 micrograms.ml-1), but that the SP-B variant [R/K-->S]SP-B1-78 was less potent (MIC = 500 micrograms.ml-1). PMID- 9392849 TI - Staurosporine-dependent activation of human endothelial cell monolayers for neutrophil adherence by secretoneurin. AB - Functions of secretoneurin include chemotaxis for monocytes and endothelial cells, and inhibition of endothelial cell proliferation. Inhibition of monocyte chemotaxis by staurosporine indicated involvement of specific signaling mechanisms. We have tested effects of kinase inhibitors on activation of endothelial cells for neutrophil adherence by secretoneurin. Pretreatment of endothelial cells by secretoneurin induced in endothelium increased adhesiveness to neutrophils. Addition of staurosporine, an inhibitor of protein kinase C, completely abolished endothelial cell activation, whereas tyrphostin-23, a tyrosin kinase inhibitor, had no effect. Results on activation of neutrophil endothelial cell adherence by secretoneurin demonstrate that specific signaling mechanisms are involved in endothelial cell activation. PMID- 9392850 TI - Endothelin-1 induces bronchial myofibroblast differentiation. AB - Endothelin-1 may contribute to bronchial smooth muscle constriction and airway remodelling in asthma, where bronchial epithelial cells represent an important source of this peptide. We report here that asthmatic bronchial epithelial cells exposed to allergens in vitro induce the differentiation of airway fibroblasts into myofibroblasts, and that they do so through a granulocyte/macrophage colony stimulating factor-mediated upregulation of endothelin-1 production. By this mechanism bronchial epithelial cells may participate in the genesis of bronchial subepithelial fibrosis, a process which contributes to airway narrowing in asthma. PMID- 9392851 TI - Rat corpus luteum expresses both PACAP and PACAP type IA receptor mRNAs. AB - Both pituitary adenylate cyclase-activating polypeptide (PACAP) and PACAP type I receptor gene expressions were detected in the corpus luteum of pregnant mare's serum gonadotropin (PMSG)-human chorionic gonadotropin (hCG)-treated immature rats using reverse transcription-polymerase chain reaction (RT-PCR). RT-PCR products of the poly(A)+ RNA extracted from rat corpora lutea yielded dominant DNA bands that corresponded to segments of PACAP mRNA (453 bp) and PACAP type IA receptor mRNA (290 bp) spanned by the PCR primers. The identities of the PACAP cDNA and the PACAP receptor cDNA fragments were confirmed by Southern blot hybridization analyses. Our results showed that PACAP mRNA and PACAP type IA receptor mRNA are synthesized within luteinized cells of rat ovary, and suggest that PACAP is closely linked to the reproductive process. PMID- 9392852 TI - Pro-dopamine effects of neurotensin on sensorimotor gating deficits. AB - Neurotensin is a neuropeptide which coexists with mesolimbic dopamine and has exhibited neuroleptic-like activity in the nucleus accumbens. This study examined the effects of neurotensin infused into the nucleus accumbens on prepulse inhibition (PPI) of the rat's acoustic startle reflex, a measure which is relevant to the sensorimotor gating deficits seen in schizophrenia. Neurotensin (5 micrograms) had no effect on the amplitude of the acoustic startle reflex nor on baseline PPI, but it potentiated the disruption of PPI produced by amphetamine and apomorphine. This is the first report of a pro-dopamine action for intra accumbens neurotensin, and suggests that a complex behavioral pharmacology is associated with this neuropeptide. PMID- 9392853 TI - CGRP-beta unlike CGRP-alpha has no osteogenic stimulatory effect in vitro. AB - The purpose of this study is to test whether CGRP-beta has an osteogenic stimulating effect, as does CGRP-alpha, on rat bone marrow cells in vitro. CGRP beta in different doses was added daily to bone marrow white cells, which were harvested from rats, then seeded onto a previously prepared layer of fibroblasts. CGRP-alpha in different doses was used as a positive control. Fourteen days after the start of the experiment, there was no statistical difference in the number of bone colonies between the control and CGRP-beta dishes. The CGRP-alpha dishes demonstrated an increase in the number of colonies with an increase of peptide dose. PMID- 9392854 TI - Gastrin-releasing peptide1-27, unlike bombesin, does not reduce sham feeding in rats. AB - We compared the potencies of systemic administration of bombesin (BN) and its mammalian homologue gastrin-releasing peptide (GRP) to decrease sham feeding in rats. Bombesin (at doses of 8, 16 and 32 micrograms/kg, intraperitoneally) inhibited sham feeding by 37% (p < 0.001), 58% (p < 0.001) and 65% (p < 0.001), respectively, confirming previous results. Gastrin-releasing peptide (16, 32, and 64 micrograms/kg) failed to affect sham feeding. Bombesin (16 micrograms/kg) and gastrin-releasing peptide (32 micrograms/kg) inhibited real feeding by 64% (p < 0.001) and 44% (p < 0.004), respectively. Pregastric food stimulation is not sufficient for the inhibitory action of GRP. PMID- 9392855 TI - Homology modeling of glycosyl hydrolase family 18 enzymes and proteins. AB - Using state-of-the-art homology modeling methods, three-dimensional coordinates for three family 18 glycosyl hydrolases were determined. The structures for Gp39, Brp39, and chitotriosidase were computer determined using the X-ray coordinates from SmChiA. The results of the modeling efforts are assessed, and comparison of the modeled structures to other known family 18 members is made. PMID- 9392856 TI - Modeling studies of binding of sea raven type II antifreeze protein to ice. AB - Certain plants, insects, and fish living in cold environments prevent tissue damage due to freezing by producing antifreeze proteins or antifreeze glycoproteins that inhibit ice growth below the normal equilibrium freezing point of water in a noncolligative fashion. In polar fish these macromolecules, taking into account their structural characteristics, are grouped into three broad classes, namely Type I, Type II, and Type III. In this paper we report the results of our studies on the stereospecific binding of sea raven, a Type II antifreeze protein (AFP) to (111) hexagonal bipyramidal faces of ice. Earlier studies of Type I and Type III AFPs have shown that stereospecific binding of these proteins, recognizing specific planes of ice, is essential for their noncolligative antifreeze point depression. Moreover, as it has been shown for the AFT of Type I, this binding also occurs along specific vectors on these planes and also is enantioselective, distinguishing between the mirror related directions. In this study we will show, by using molecular modeling, that the fold of Type II AFP could facilitate a stereospecific mode of interaction with (111) planes of ice. Similar to Type I AFP, preferential directionality of binding was also observed in the simulations. PMID- 9392857 TI - Structural characterization of the molecular dimer of the peptide antibiotic vancomycin by distance geometry in four spatial dimensions. AB - The conformation of a dimer of the peptide antibiotic vancomycin is developed from computer simulations based on experimental distance constraints derived from high-resolution NMR measurements. The conformation and topological array of the dimer are determined by a distance geometry based method using the molecules cast into four spatial dimensions. This method was imperative for the refinement of vancomycin given the entwining of the monomers (including hydrogen bonding and interactions between the aromatic ring systems) within the dimer. The development of the high-resolution structure of the monomer and then simple molecular modeling to create the dimer which fulfills all of the experimental observations was not possible. In contrast, the refinement protocol using the dimer cast into four spatial dimensions was able to quickly locate conformations in which both the intra- and intermolecular nuclear Overhauser effects were satisfied. These structures, once converted back to three dimensions, were further refined using standard molecular mechanics energy minimization. The structural characteristics of the dimer with respect to binding to the cell-wall precursor are described. PMID- 9392858 TI - Using neural network predicted secondary structure information in automatic protein NMR assignment. AB - In CAPRI, an automated NMR assignment software package that was developed in our laboratory, both chemical shift values and coupling topologies of spin patterns are used in a procedure for amino acids recognition. By using a knowledge base of chemical shift distributions of the 20 amino acid types, fuzzy mathematics, and pattern recognition theory, the spin coupling topological graphs are mapped onto specific amino acid residues. In this work, we investigated the feasibility of using secondary structure information of proteins as predicted by neural networks in the automated NMR assignment. As the 1H and 13C chemical shifts of proteins are known to correlate to their secondary structures, secondary structure information is useful in improving the amino acid recognition. In this study, the secondary structures of proteins predicted by the PHD protein server and our own trained neural networks are used in the amino acid type recognition. The results show that the predicted secondary structure information can help to improve the accuracy of the amino acid recognition. PMID- 9392859 TI - Using artificial neural networks to classify the activity of capsaicin and its analogues. AB - Back-propagation artificial neural networks (ANNs) were trained with parameters derived from different molecular structure representation methods, including topological indices, molecular connectivity, and novel physicochemical descriptors to model the structure--activity relationship of a large series of capsaicin analogues. The ANN QSAR model produced a high level of correlation between the experimental and predicted data. After optimization, using cross validation and selective pruning techniques, the ANNs predicted the EC50 values of 101 capsaicin analogues, correctly classifying 34 of 41 inactive compounds and 58 of 60 active compounds. These results demonstrate the capability of ANNs for predicting the biological activity of drugs, when trained on an optimal set of input parameters derived from a combination of different molecular structure representations. PMID- 9392860 TI - Distribution analysis of the variation of B-factors of X-ray crystal structures; temperature and structural variations in lysozyme. AB - The B-factor (isotropic temperature factor) data for X-ray structures of hen egg white lysozyme from the study of Young et al. (Young, Dewan, Nave, and Tilton J. Appl. Cryst. 1993, 26, 309-319) potentially contain information about the relative contributions of static and dynamic variation to these factors. The six structures of the protein were obtained at two widely different temperatures (100 and 298 K), with two crystal forms (monoclinic and tetragonal) and other experimental differences. In addition, the monoclinic lysozyme crystals with two molecules per asymmetric unit allow direct examination of variation between structures determined under identical conditions at both temperatures. The B factors from these structures all have complex distribution functions as might be expected considering all of the influences that these values must reflect. The empirical cumulative distribution functions (eCDF's) of these data show that they are representative of complex, multicomponent distributions. Distribution analysis using the DANFIP procedure (Wampler, Anal. Biochemistry 1990, 186, 209 218) of the data sets reveals that they can be modeled as four to six Gaussian subpopulations, that these subpopulations do not correlate with specific atom types, specific amino acid residues or fixed locations in the structure. While they do seem to correlate with localized groupings of atoms, these grouping vary from structure to structure even within the same crystal under the same conditions. Temperature seems to have a global effect in this case, but it is clear that other factors including experimental error influence the distribution of B-factors within a given structure. This analysis also helps explain the oft observed lack of atomic level correlation between experimental B-factors and calculated mean square displacements from molecular dynamics simulations. PMID- 9392861 TI - Intracellular transmission in cell volume regulation in Ehrlich ascites tumor cells. PMID- 9392863 TI - Role of actin filament organization in cell volume and ion channel regulation. AB - The actin cytoskeleton is an intracellular structure, which is involved in the onset and control of cell shape and function. In order for this relevant network to control its own and thus cell volume, specific interactions between the actin cytoskeleton and ion channel regulation controlling intracellular salt and water homeostasis may be invoked. The hypotonic shock-induced, cell volume regulatory decrease (RVD) of most eukaryotic cells is a particularly useful example, as it is initiated and regulated by concerted processes involving both adaptive changes in actin filament organization and bulk fluid extrusion triggered by saline movement and the consequent decrease in cell water. The onset of RVD is linked to the selective activation of osmotically-sensitive ion channels and other relevant ion transport mechanisms involved in the net ionic movement from the cytosol. Such regulatory processes, entailing effector changes in actin filament organization which target the plasma membrane, are largely unknown. In this report, recent studies are summarized implicating dynamic changes in gel properties of the actin cytoskeleton as the effector mechanism in the regulation of ion channel activity, and thus cell volume, in human melanoma cells. Based on the characterization of the hypotonic cell volume regulatory response of human melanoma cells devoid of a functional actin-binding protein (ABP-280, a filamin homolog) and their genetically rescued counterpart transfected with a functional ABP, a hypothesis is raised which is consistent with a regulatory "sensory" mechanism based on the ability of actin networks to respond to changes in the intracellular water-salt homeostasis, which in turn effects signals controlling membrane function, including ion channel activity. PMID- 9392862 TI - Confocal microscopic observation of cytoskeletal reorganizations in cultured shark rectal gland cells following treatment with hypotonic shock and high external K+. AB - The dogfish shark (Squalus acanthias) rectal gland (SRG) cell has served as a model experimental system for investigating the relationship between the actin cytoskeleton and cell volume regulation. Previous reports employing conventional fluorescence microscopy of tissue slices have shown that cells exposed to high external K+ and hypotonically-induced cell swelling displayed a fading of F-actin staining intensity, particularly at the basolateral cell borders. However, spectroscopic measurement of the F-actin present in similarly treated rectal gland slices failed to demonstrate a net change in F-actin amount. In an effort to resolve the structural reorganizations of F-actin which may be occurring during high K+ and hypotonic shock treatments, we have used cultured SRG cells in conjunction with confocal microscopic immunocytochemical localization techniques to examine actin filament, microtubule, and cytokeratin filament dynamics under these two experimental conditions. The results reveal that F-actin in control cells exists in an array of parallel linear bundles (which do not appear to be stress fiber-like given their lack of staining for myosin II or alpha-actinin) that is reorganized to a punctate pattern in hypotonic shock and a dense meshwork in high K+. The linear bundle pattern of F-actin returns in cells undergoing regulatory volume decrease. Quantitative western blotting of F-actin in SRG cell detergent extracted cytoskeletons indicates no significant difference in the relative amounts of F-actin present in control, hypotonic shocked, or high K+ cells. Anti-tubulin and anti-cytokeratin labeling of the treated SRG cells suggest that these other major cytoskeletal elements are not significantly altered by the treatments. Taken together, our results reinforce the concept that there is an association between the structural organization of the actin cytoskeleton and cell volume regulation in the SRG epithelial cells. PMID- 9392864 TI - Biochemistry and physiology of carbohydrates in the renal collecting duct. AB - Using 13C-NMR analysis of cell extracts, enzymatic determination of metabolites and cofactors as well as enzyme assays on cell homogenates aerobic and anaerobic glycolysis, sorbitol formation by aldose reductase, the pentose phosphate shunt, and gluconeogenesis could be identified as the major pathways of D-glucose metabolism in renal inner medullary collecting ducts. In flux studies it was shown that D-glucose enters the collecting duct cells via a sodium-independent, cytochalasin- and phloretin-inhibitable transport system located at the basal lateral cell side. At the same side sorbitol leaves the cells during regulatory volume decrease in a calcium-calmodulin-dependent fashion. From cell isolation studies it is proposed that sorbitol is taken up by adjacent (interstitial) cells, converted into fructose and then recycled to the collecting duct cells. This cycle might prevent carbohydrate wasting. Thus, IMCD cells exhibit unique aspects of carbohydrate biochemistry and physiology which enable them to function in a surrounding of low oxygen tension, low substrate supply, and extreme changes in extracellular osmolality. PMID- 9392865 TI - Characterization and regulation of H-K-ATPase in intercalated cells of rabbit cortical collecting duct. AB - K-dependent H+ extrusion was investigated using fluorescence techniques in rabbit cortical collecting tubules (CCTs). Experiments were performed in split-open tubules from normal animals exposed to the intracellular pH indicator 2',7' bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF). This preparation permitted the study of individual intercalated cells (ICs). In the ICs, partial recovery of pH(i) was observed in response to an acute acid load upon readdition of 5 mM K to the superfusate. This recovery was SCH 28080-inhibitable (10(-5) M) and ouabain insensitive suggesting the process is mediated by a gastric-type H-K ATPase. To see if H-K ATPase plays a role in acid secretion its function was evaluated under chronic metabolic acidosis (CMA) conditions. CMA was induced by replacing drinking water with 75 mM NH4Cl in 5% sucrose for 10-14 days. The SCH 28080 inhibitable K-dependent pH(i) recovery rate was three-fold higher in CMA ICs compared to controls. To determine the location of the H-K ATPase, CCTs were microperfused and individual peanut lectin binding (PNA) ICs studied. K-dependent pH(i) recovery was measured in response to an NH4Cl pulse. An apical SCH 28080 inhibited K-dependent pH(i) recovery process was observed in control and CMA ICs. Taken together these data confirm the existence of a gastric-type H-K ATPase in ICs of rabbit CCT. Based on our findings the H-K ATPase is found on the apical side of the cell and is stimulated under conditions of CMA. PMID- 9392866 TI - Volume-activated osmolyte channel in skate erythrocytes: inhibition by pyridoxal derivatives. AB - Volume expansion of erythrocytes of little skate, Raja erinacea, triggers the opening of an osmolyte channel. We review this transport mechanism and further investigate the channel's physicochemical nature by probing the channel with a series of pyridoxine derivatives in skate RBC as well as in epithelial cells: MDCK and C6 glioma cells and in skate hepatocytes. The identity of the transport mechanism (band 3 vs. an anion channel) which mediates the swelling-activated efflux of osmolytes in fish RBC is controversial. Therefore, we compared taurine and Cl- effluxes in similar conditions. We found that there is significant Cl- loss from volume-expanded skate RBC. However, there was no effect of either hypotonicity or a number of taurine transport inhibitors on this loss. Utilizing changes in intracellular pH as a means of indirectly measuring H+/Cl- cotransport, we found that a rise in cell pH accompanied the loss of Cl-. This suggests that Cl- efflux could occur via a H+/Cl- cotransporter. To probe and compare the osmolyte channel (taurine efflux) of the skate RBC and three other cell types we used a family of pyridoxine inhibitors. The inhibitory patterns for the skate erythrocytes and hepatocytes differed from those for MDCK and C6 glioma cells and the two former cell types differed from each other. Therefore, the results show that the osmolyte channel in the skate differs from that in other epithelial cells with regard to pyridoxine derivative binding properties. PMID- 9392867 TI - Dual pathways for organic anion secretion in renal proximal tubule. AB - Transport on the "classical" organic anion system in renal proximal tubule is specific, active, Na-dependent, and ouabain sensitive. Here we review recent studies using intact teleost proximal tubules and laser scanning confocal microscopy which show that the secretion of large organic anions, such as, fluorescein-methotrexate (FL-MTX, Mw 923 Da) is handled by a separate and distinct organic anion transport system. In contrast to the classical system, FL MTX uptake into cells and secretion into the tubular lumen was ouabain insensitive and largely Na-independent. KCN did not affect cellular uptake but abolished secretion into the lumen. PAH and probenecid, potent inhibitors of transport on the classical system, were weak inhibitors of FL-MTX transport. Uptake and secretion of FL-MTX were inhibited by micromolar concentrations of other organic anions (MTX, folate, bromocresol green, bromosulfonphthalein). FL MTX secretion into the lumen was inhibited by leukotriene C4 and cyclosporine A, neither of which affected transport of the model substrate for the classical system, fluorescein. Thus, FL-MTX secretion is specific, but largely Na independent and ouabain-insensitive. Both the basolateral and luminal steps in FL MTX transport differ from those associated with fluorescein and P-aminohippurate secretion. PMID- 9392868 TI - ATP regulation of a swelling-activated osmolyte channel in skate hepatocytes. AB - Hypotonic swelling of isolated skate hepatocytes activates a regulatory volume decrease (RVD) which is achieved in part by the release of taurine and other intracellular organic osmolytes. Volume-activated taurine efflux appears to be mediated by an anion channel that exhibits a taurine/chloride permeability ratio of approximately 0.2. Of significance, this channel was shown to be regulated by intracellular nucleotide. When intracellular ATP was decreased to about 50% of control levels, channel opening was completely prevented. Many putative ion channel blockers were found to inhibit the channel indirectly, by depleting intracellular ATP, rather than by directly interacting with the channel. Investigators using these channel blockers in whole cell preparations should be aware of this alternative mechanism. Cell swelling-activated taurine efflux was also inhibited by HgCl2, DIDS, and pyridoxal 5-phosphate, at concentrations of these agents that had no effect on intracellular ATP levels, suggesting additional mechanisms of inhibition and regulation of the volume-sensitive osmolyte channels. PMID- 9392869 TI - Pump-leak parallelism in sodium-absorbing epithelia: the role of ATP-regulated potassium channels. AB - In all Na(+)-absorbing and Cl(-)-secreting epithelia, an increase in the activity of the Na+,K(+)-pump at the basolateral membrane is accompanied by an increase in the K+ conductance of that barrier and vice versa. We have recently identified an ATP-regulated K+ channel, K(ATP), in basolateral membrane vesicles isolated from Necturus maculosa small intestinal epithelial cells that could be responsible for this parallelism between pump activity and leak. Thus, an increase in pump activity would result in a decrease in local ATP activity and an increase in local ADP activity and, in turn, an increase in the open-probability of the channel whereas a decrease in in pump activity would have the opposite effect. Further, the likelihood that the number of pumps far exceeds the number of leaks per unit area of membrane suggests that the ATP and ADP activities that influence K(ATP) channel activity may differ markedly from the "bulk" cytoplasmic values. PMID- 9392870 TI - Potential interplay between luminal growth factors and increased tight junction permeability in epithelial carcinogenesis. PMID- 9392871 TI - Sulfonamides and secretion of aqueous humor. AB - The connection between carbonic anhydrase and the formation of aqueous humor arose in the decade 1950-1960 when a number of experiments of differing types showed that HCO3- ion catalytically formed in the ciliary process from CO2 was a principal element in the production of the aqueous. It was soon shown that inhibitors of the enzyme, given systemically, slowed HCO3- formation and sodium and fluid transport and thereby lowered intraocular flow and pressure in normal vertebrates and in patients with glaucoma. In the past 15 years successful efforts have been made to produce sulfonamide inhibitors that reach the ciliary process after local application to the cornea. The effects of locally acting sulfonamides are largely due to their amphoteric properties. They are soluble in both lipid and non-lipid media and penetrate cornea and sclera to reach ciliary process. Potency against carbonic anhydrase II is of the order of K(I) = 10(-9) M, so that topical application inhibits essentially all enzyme in the processes and is an effective nontoxic treatment for glaucoma. PMID- 9392873 TI - Transport mechanisms that mediate the secretion of chloride by the rectal gland of Squalus acanthias. AB - The rectal gland of Squalus acanthias secretes chloride by a mechanism that has been termed "secondary active transport" because it depends on the activity of Na K-ATPase. As currently described, chloride enters the cell across the basolateral cell membrane via the 2 chloride: sodium: potassium cotransporter. The energy for this electroneutral uphill movement of chloride and potassium is provided by the gradient for sodium directed into the cell. Present in the basolateral cell membrane is Na-K-ATPase that maintains the gradient for sodium. A potassium conductance, present as well in the basolateral cell membrane, recirculates the potassium. Chloride exits the cell across the luminal membrane via CFTR, the chloride conductance. This mechanism is widely distributed throughout vertebrates. This report reviews the experimental observations that led to the current definition of the mechanism of chloride transport in the rectal gland. PMID- 9392872 TI - Potential contribution of epithelial Na+ channel to net secretion of aqueous humor. AB - The aqueous humor of the eye is secreted by the bilayered ciliary epithelium, consisting of the pigmented (PE) cell layer facing the stroma and the nonpigmented (NPE) cell layer facing the aqueous humor. Cells within each layer and between the two layers are linked by gap junctions, forming a ciliary epithelial syncytium. Unidirectional secretion from the stroma to the aqueous proceeds both through the cells (the transcellular pathway) and between the cells (the paracellular pathway). Net formation of aqueous humor must, however, be the algebraic sum of unidirectional secretion and unidirectional reabsorption from the aqueous humor back into the stoma. The mechanisms potentially underlying reabsorption of aqueous humor by the NPE cells have recently been addressed by studying the regulatory response (RVI) of anisosmotically shrunken NPE cells. The results indicated that epithelial Na+ channels with a high affinity to amiloride likely contribute to reabsorption of solute from the aqueous humor. We have substantiated this possibility by using Northern analysis to identify in human ciliary body RNA a 3.7-kb transcript corresponding to the alpha-subunit of the amiloride-sensitive, alpha beta gamma-ENaC epithelial sodium channel. We have also found that the Na(+)-channel inhibitor benzamil inhibits the RVI without affecting the cell volume of isotonic cell suspensions. This observation supports the hypothesis that the low conductance, highly selective epithelial Na+ channel is activated by shrinkage and contributes to unidirectional reabsorption as aqueous humor. Examples are provided of how the integrative regulation of aqueous humor formation can involve conjugate actions on both unidirectional secretion and reabsorption. PMID- 9392874 TI - Cadmium disrupts the signal transduction pathway of both inhibitory and stimulatory receptors regulating chloride secretion in the shark rectal gland. AB - The heavy metal cadmium causes nephrotoxicity and alters the transport function of epithelial cells. In the shark rectal gland, chloride secretion is regulated by secretagogues and inhibitors acting through receptors coupled to G proteins and the cyclic AMP-protein kinase A pathway. We examined the effects of cadmium on the response to the inhibitory peptide somatostatin (SRIF), and to the stimulatory secretagogues forskolin and vasoactive intestinal peptide (VIP). In control experiments, SRIF (100 nM) entirely inhibited the chloride secretory response to 10 microM forskolin (maximum chloride secretion with forskolin 1984 +/- 176 microEq/h/g; with forskolin + SRIF 466 +/- 93 microEq/h/g, P < 0.001). Cadmium (25 microM) entirely reversed the inhibitory response to SRIF (chloride secretion 2143 +/- 222 microEq/h/g) and caused an overshoot (2917 +/- 293 microEq/h/g) that exceeded the response to forskolin (P < 0.01). Cadmium also enhanced forskolin-stimulated chloride secretion (2628 +/- 418 vs. 1673 +/- 340 microEq/h/g, P < 0.02) and reversed the declining phase of the forskolin response. Cadmium had a concentration-dependent, biphasic effect on the response to VIP. Cd (10-100 microM) increased both chloride secretion and tissue cyclic AMP content, whereas higher concentrations (1 mM) inhibited chloride secretion and cyclic AMP accumulation. Our findings provide evidence that Cd disrupts the signal transduction pathways of both inhibitory receptors and secretagogues regulating cAMP mediated transport in an intact epithelia. The results are consistent with direct effects of cadmium on adenylate cyclase and/or phosphodiesterase activity in this marine epithelial model. PMID- 9392875 TI - New protopine and benzyltetrahydroprotoberberine alkaloids from Aristolochia constricta and their activity on isolated guinea-pig ileum. AB - Five new protopine-type alkaloids, 3,5-di-O-methylconstrictosine (1), 5,6-dihydro 3,5-di-O-methylconstrictosine (2), 5,6-dihydroconstrictosine (3), constrictosine (4), 3-O-methylconstrictosine (5), and a novel 8-benzylberberine-type alkaloid, ( )-8 beta-(4'-hydroxybenzyl)-2,3-dimethoxyberbin-10-ol (6) were isolated from the aerial parts of Aristolochia constricta. Their structures were elucidated by physical and spectroscopic data. The results of our pharmacological experiments indicated that MeOH extract, its partially purified fraction VI and the protopine derivatives constrictosine 1-5, significantly reduced, in a dose dependent manner, the electrical, acetylcholine, and histamine contractions of the isolated guinea-pig ileum. PMID- 9392876 TI - Triterpenoid saponins from Trevesia sundaica. AB - Six new bisdesmosidic saponins 1-6, along with four known triterpenoid saponins were isolated from the aerial parts of Trevesia sundaica (Araliaceae). Their structures were determined by 1H-1H correlation spectroscopy (COSY, TOCSY, ROESY) and 1H-13C heteronuclear correlation (HSQC, HMBC) NMR experiments, FABMS, and chemical data. PMID- 9392877 TI - Isolation, synthesis, and antiplatelet aggregation activity of resveratrol 3-O beta-D-glucopyranoside and related compounds. AB - Resveratrol 3-O-beta-D-glucopyranoside (1) has been isolated from the seeds of Erythrophleum lasianthum (Caesalpinioidae, Leguminosae), a South African plant used in traditional medicine, and has shown antiplatelet aggregation activity. The synthesis of 1, related hydroxystilbenes, and their glucosides has been undertaken to provide larger quantities, for further biological evaluation, and has been accomplished via Wittig reactions followed by glucosylation under phase transfer catalysis. PMID- 9392878 TI - Isolation of a novel Kunitz family protease inhibitor in association with Tethya hemolysin from the sponge Tethya ingalli. AB - Aqueous extracts from the New Zealand sponge Tethya ingalli (Hadromerida) displayed potent cytotoxicity in the NCI's 60-cell-line human tumor panel. Fractionation of the extract by ammonium sulfate precipitation, gel filtration, ultrafiltration, and both hydrophobic interaction and reversed-phase chromatography resulted in the isolation of two biologically active proteins. The first protein, Tethya protease inhibitor (TPI), which was purified to homogeneity, inhibited trypsin with an EC50 of 65 nM. TPI had a molecular mass of 11,431 Da, and an isoelectric point of 8.2. A partial N-terminal amino acid sequence determined for TPI showed significant homology with protease inhibitors of the Kunitz family. The second isolated protein displayed potent cytotoxicity, with pronounced selectivity for certain tumor cell lines (e.g., ovarian, renal, CNS, and breast). The latter protein, which had an apparent molecular weight of 21 kDa (SDS-PAGE), also lysed human red blood cells (EC50 of 39 nM) and was similar to a hemolysin previously isolated from the sponge Tethya lycinurium. PMID- 9392879 TI - Antitumor agents. 180. Chemical Studies and cytotoxic evaluation of cumingianosides and cumindysoside A, antileukemic triterpene glucosides with a 14,18-cycloapotirucallane skeleton. AB - Treatment of cumingianosides and cumindysoside A, which possess a 14,18 cycloapotirucallane skeleton, with p-toluenesulfonic acid in CH2Cl2 yielded new triterpene glucosides. Cumingianoside A (1) gave 10 and 11, along with cumingianoside Q (5). The structures of 10 and 11 were determined on the basis of spectral examination and contained a dammar-13(17)-ene and a 17(R),23(R) epoxydammarane skeleton, respectively. Cumingianoside C (2) afforded, together with cumingianoside P (6), products 12 and 13, which were similar to 10 and 11, respectively. With a short reaction time at room temperature, cumingianoside E (3) yielded cumingianoside D (4). In contrast, when 3 was treated with p toluenesulfonic acid in CH2Cl2 overnight at 5 degrees C, it gave two products, 9 and 14. Extensive spectroscopic examination revealed that 9 possessed a dammar-12 ene skeleton, while 14 was a pentacyclic tetranortriterpene glucoside with a novel skeleton. Cumindysoside A (8) gave a product (15) similar to 14. The cytotoxicities of 9-15 were evaluated against a panel of 58 human tumor cell lines. Compounds 11-15 exhibited potent cytotoxicity with log GI50 values ranging from -7.11 to -4.94, especially against leukemia and colon-tumor cell lines. PMID- 9392880 TI - Sesquiterpenoids of the drimane class from a sponge of the genus Dysidea. AB - Ten sesquiterpenoids, including seven new ones, have been isolated from an undescribed sponge of the genus Dysidea. Compounds 1-8 are sesquiterpenoids of the drimane class, while 9 and 10 are 12-norsesquiterpenoids of the same structural class. The structures of novel compounds have been determined by combined spectroscopic methods. These compounds exhibited moderate antimicrobial and enzyme inhibitory (Na+/K(+)-ATPase and PLA2) activities. PMID- 9392882 TI - New cembranoid diterpenes and a geranylgeraniol derivative from the common Caribbean sea whip Eunicea succinea. AB - A recent collection and extraction of the common Caribbean sea whip Eunicea succinea from Puerto Rico has produced four previously undescribed representatives of the cembrane family of diterpenes (2, 3, 4, and 6). A new geranylgeraniol derivative, 8, was also isolated as a minor constituent. The chemical structures of the new compounds were carefully established by spectroscopic and chemical methods in addition to detailed NMR spectral comparisons with known cembranoid models from Eunicea. PMID- 9392881 TI - Oxidations of vincristine catalyzed by peroxidase and ceruloplasmin. AB - The dimeric Catharanthus alkaloid vincristine (1) is oxidized to the same ring fission product in incubations with either horseradish peroxidase or the human serum copper oxidase ceruloplasmin. Horseradish peroxidase-catalyzed oxidation of vincristine requires hydrogen peroxide, whereas ceruloplasmin-catalyzed oxidation of vincristine requires chlorpromazine as a "shuttle oxidant". Preparative-scale incubations allowed for the production, isolation, structural characterization, and biological evaluation of the metabolite. The metabolite was identified as the heterocyclic ring cleavage product N-formylcatharinine (5). N-Formylcatharinine was 118 times less active than vincristine in an in vitro test against a human T cell leukemic cell line. Therefore, these enzyme-catalyzed reactions lead to bioinactivation of vincristine. PMID- 9392883 TI - Zanhasaponins A and B, antiphospholipase A2 saponins from an antiinflammatory extract of Zanha africana root bark. AB - A MeOH extract from Z. africana was examined for topical antiinflammatory activity and proved to be active against arachidonic acid (AA) acute edema, 12-O tetradecanoylphorbol 13-acetate (TPA)-induced chronic inflammation, and oxazolone delayed-type hypersensitivity in mice. The extract also showed significant inhibitory activity of Naja naja phospholipase A2 when a polarographic method was used. Two oleanane-type triterpene saponins, zanhasaponins A (1) and B (2), and the cyclitol pinitol (4), isolated from the extract, were active as inhibitors of PLA2. A further saponin, zanhasaponin C (3) was inactive in this assay. PMID- 9392884 TI - Saniculoside N from Sanicula europaea L. AB - Extracts from the aerial parts of Sanicula europaea L. were investigated for their anti-HIV activity, and the 50% ethanolic extract was shown to exhibit the highest activity. A new triterpene saponin glycoside, 21 beta-(angeloyloxy)-3-O [beta-D-arabinopyranosyl(1-->4)-beta- D-glucopyranosyl (1-->3)-beta-D glucuronopyranosyl propyl ester]-3 beta,15,16,22 alpha,28 beta-pentahydroxy delta(12)-oleanene, saniculoside N (1), in addition to the known phenolic acids, rosmarinic acid (2), and caffeic acid (3) were isolated as major components. Rosmarinic acid was established as the principal active substance. PMID- 9392885 TI - A new steroidal glycoside from a Caribbean gorgonian, Eunicea sp.1. AB - A new saponin possessing a pregnene-derived aglycon (1) has been isolated from the Caribbean gorgonian octocoral Eunicea sp. The structure of the new compound was assigned on the basis of chemical and spectral studies. PMID- 9392886 TI - Multidrug-resistance modulators from Stephania japonica. AB - An alkaloidal extract of the vines of Stephania japonica showed multidrug resistance-reversing activity as demonstrated by the bicinchoninic acid assay. Two known bisbenzylisoquinoline alkaloids, isotrilobine (1) and trilobine (2), were isolated by bioassay-directed fractionation and separation. Isotrilobine (1) was shown to be as active as verapamil (3) in reversing doxorubicin resistance in human breast cancer cells. PMID- 9392887 TI - Four new cytotoxic germacranolides from Carpesium divaricatum. AB - In a bioassay-guided search for cytotoxic compounds from higher plants of South Korea, four new sesquiterpenes of the germacranolide type, named cardivins A (1), B (2), C (3), and D (4), have been isolated from the aerial parts of Carpesium divaricatum. Structures of these compounds were elucidated on the basis of spectroscopic techniques. Compounds 1, 2, 3, and 4 showed cytotoxicity to the human tumor cells, A-549 (nonsmall cell lung), SK-OV-3 (ovary), SK-MEL-2 (skin), XF-498 (central nervous system), and HCT-15 (colon). PMID- 9392888 TI - New phomopsolides from a Penicillium sp. AB - Investigation of the bioactive compounds from a Penicillium sp. isolated from the inner bark of the Pacific yew, Taxus brevifolia, led to the isolation of the known furanone 1, and a series of phomopsolides. The phomopsolide fractions contained phomopsolides A and B, which have previously been described, and three new phomopsolides. The structures of the new phomopsolides were deduced by comparison of their NMR spectra to those of the known compounds. PMID- 9392890 TI - Values of hypercompetitive and personal development competitive individuals. AB - The value systems of hypercompetitive and personal development competitive individuals were examined in a sample of university undergraduates. As expected, people higher in hypercompetitiveness and in personal development competitiveness were both more likely to endorse values related to self-contained individualism such as achievement, hedonism, and a striving for an exciting and challenging life, but only hypercompetitives endorsed the value of power and control over others. Moreover, the data indicated that people higher in personal development competitiveness were more prone to endorse values related to ensembled individualism. In particular, they strongly endorsed values associated with social concern, that is, with caring about the well-being of others and with treating them with respect and as equals, whereas hypercompetitives expressed a lack of such concern. Discussion centered on the socialization process and how it can foster the development of different competitive orientations. PMID- 9392889 TI - Antimycobacterial polyynes of Devil's Club (Oplopanax horridus), a North American native medicinal plant. AB - Two new (3 and 5), as well as three known (1, 2, and 4), polyynes were isolated from Devil's Club (Oplopanax horridus; Araliaceae), a medicinal plant of North America. The structures were established by 1H and 13C NMR. The absolute configurations of 2 and 5 were determined by application of Mosher's method. All the polyynes exhibited significant anti-Candida, antibacterial, and antimycobacterial activity, with an ability to kill Mycobacterium tuberculosis and isoniazid-resistant Mycobacterium avium at 10 micrograms/disk in a disk diffusion assay. PMID- 9392891 TI - Confirmatory factor analysis of the personality disorder subscales from the Inventory of Interpersonal Problems. AB - The purpose of this study was to identify the best fitting hierarchical factor structure of the subscales for personality disorders developed from the Inventory of Interpersonal Problems (IIP). In earlier work, 5 subscales associated with a diagnosis of personality disorder (PD) had been developed. With data collected from 5 additional samples at 4 sites (N = 1004), relations among the IIP-PD subscales were investigated using confirmatory factor analysis. Several competing models were tested, and a second-order model with 1 second-order factor and 5 first-order factors provided the best fit to the data. The results support the hypothesis of a single latent construct reflecting general personality dysfunction. Measures of this construct may be useful for screening patients into yes versus no PD groups. PMID- 9392893 TI - Stability of a Q-sort model of optimal mental health. AB - This study explores clinicians' conceptions of optimal mental health as a function of target age and asks whether contemporary conceptions of optimal mental health differ from Block's (1961) earlier Q-sort model. Ten experienced clinicians from Northwestern University described the personality characteristics of optimally adjusted 25- and 50-year-old targets using the California Q-set. Item scores from these Q-sorts were aggregated to form 25-year-old, 50-year-old, and composite templates of optimal mental health. Using the Spearman-Brown formula, the reliability of the 10-judge composite was .97. The 3 templates were very highly correlated with each other and with Block's original template. Q-sort descriptions of optimal mental health are remarkably stable over 2 generations of clinicians and between young adulthood and mid-life. PMID- 9392892 TI - Relations of five-factor model antagonism facets with personality disorder symptomatology. AB - The Five-Factor Model of Personality (FFM) has been used to conceptualize personality disorders as maladaptive variants of normal personality traits. This study assessed the convergence of 6 lower order traits, or facets, of FFM agreeableness versus antagonism (trust, straightforwardness, altruism, compliance, modesty, and tender-mindedness) with antisocial, borderline, narcissistic, paranoid, and passive-aggressive personality traits. Interview based scores for all of the antagonism facets except compliance demonstrated the expected relations with these personality disorder traits. Results for self reported facet scores were less clearly supportive, only yielding convergence for straightforwardness and altruism with respect to antisocial traits. It is suggested that future investigations of the FFM, or other normal personality trait models, and personality disorder symptomatology include analyses at the lower order trait level. PMID- 9392894 TI - Comparison of the big-five factor structure across samples of Chinese and American adults. AB - We compared the factor structure of Goldberg's (1992) 50-item Bipolar Rating Scale (50-BRS) in samples of Chinese (n = 198) and American (n = 303) students. Results confirmed the hypothesized five-factor pattern for the U.S. sample, and a simultaneous multisample confirmatory factor analysis showed that the same five factor pattern fit the item covariances in the Chinese sample. High levels of internal consistency were found within each sample, and a high degree of congruency of corresponding item factor loadings was obtained across samples. Overall, results supported the potential utility of the Five-Factor Model and the 50-BRS for assessing personality dimensions in Chinese culture. PMID- 9392895 TI - Manic indices on the Rorschach. AB - This study examined signs of mania on the Rorschach, specifically whether manic inpatients (n = 24) produce different thematic content and thought disorder than comparison groups of paranoid schizophrenic (n = 27) and schizoaffective (n = 25) inpatients. Rorschach protocols were scored by a trained rater for the Thought Disorder Index and the Schizoid-Affective Rating Scale. Results indicated that all 3 groups had moderate levels of thought disorder, but the manic inpatients produced significantly more combinatory thinking and affective content responses than the other 2 groups. The paranoid schizophrenic and schizoaffective patients did not produce significantly more schizoid content and were not different on any other types of thought disorder than the manic patients. These findings are discussed in terms of the contribution of thought disorder and affective thematic content in making the diagnosis of mania on the Rorschach. PMID- 9392896 TI - Motivational distortion of the 16PF by welfare recipients. AB - The effectiveness of the Sixteen Personality Factor (16PF) motivational distortion correction procedures was investigated with a sample of 212 welfare recipients who completed the 16PF while participating in a mandatory welfare-to work program. A multiple regression analysis showed that the motivational distortion (MD) score was significantly related to most of the preselected Personality factors. The regression analysis also revealed that primary E (Dominance) was associated with MD, although the manual does not require MD adjustments for this factor. Based on comparisons of mean differences at the various MD correction levels, findings indicated general support for the MD correction procedures described in the manual; however, the magnitude of the correction procedures should be used cautiously as this may overcorrect for MD on some of the 16PF primaries. The relevance of the findings also are discussed in terms of evidence for Cattell's (1968, 1973, 1986) trait-view theory as it applies to response distortions. PMID- 9392897 TI - Self-report differentiation of anxiety and depression in chronic pain. AB - The psychometric distinctiveness of self-reported anxiety and depression in patients with chronic pain was investigated. The item-level responses of 220 patients with heterogeneous pain conditions from the Beck Depression Inventory and State--Trait Anxiety Inventory State--Anxiety scale were submitted to common factor analysis. Three first-order factors were identified: depression, anxiety absent, and anxiety-present. One second-order factor of negative affect was also identified. Correlations of first-order factor scores with other psychometric measures suggested only minor distinctiveness. The findings indicated that it is possible to distinguish anxiety and depression psychometrically in patients with chronic pain but suggested that negative affect may be the primary underlying construct of the affective experience of these patients. PMID- 9392899 TI - Second-order motion perception in peripheral vision: limits of early filtering. AB - Spatial and temporal analysis of contrast-modulated sine-wave gratings reveals that the second-order motion stimulus contains two sidebands, with equal energy but moving in opposite directions, flanking a stationary carrier. Any early linear spatial filtering process in the visual system that attenuates one sideband more than the other will be detrimental to the balance between the two sidebands, so that the perceived direction of the carrier might be opposite to that of the envelope motion. We tested this hypothesis by using contrast modulated gratings presented centrally or at 20 deg in the horizontal nasal field with a two-alternative forced-choice staircase paradigm. We found that when the envelope frequency was close to that of the carrier, a second-order stimulus whose envelope motion direction was correctly identified in the fovea appeared to drift in the opposite direction in the periphery. Further increasing the envelope spatial frequency resulted in a reversed motion percept in both central and peripheral viewing conditions. For subjects to identify correctly the direction of motion of the envelope, the spatial frequency ratio of the carrier to the envelope had to be more than 2 in the fovea and more than 6 in the periphery. These phenomena in second-order motion perception can be explained by a linear model of motion detection with an early spatial filtering process. Further experiments and computer simulation show that undersampling of the carrier has little effect on second-order motion perception in the periphery, as long as the carrier is detectable. PMID- 9392898 TI - A hostility scale for the California Psychological Inventory: MMPI, observer Q sort, and big-five correlates. AB - Using two samples, we developed and validated a hostility scale that can be scored from the California Psychological Inventory (CPI) and serves as an alternate for the Cook-Medley Hostility Scale (Ho; Cook & Medley, 1954). The CPI Hostility (H) scale consists of 33 items that are either duplicates or close equivalents of specific Ho items, and the two scales correlate at least .90 in samples differing in sex. The H and Ho scales show a similar pattern of correlations with conceptually relevant MMPI scales and with observer-rated personality attributes tapping Barefoot, Peterson, et al.'s (1991) five hostility categories of Hostile Affect, Cynicism, Aggressive Responding, Social Avoidance, and Hostile Attributions. These findings provide evidence for the equivalence of the two hostility scales, as well as external validation for those personality characteristics that are purported to underlie the construct of hostility as tapped by both the original Ho scale and the new CPI H scale. PMID- 9392900 TI - Geometry of shadows. AB - Shadows provide a strong source of information about the shapes of surfaces. We analyze the local geometric structure of shadow contours on piecewise smooth surfaces. Particular attention is paid to intrinsic shadows on a surface: that is, shadows created on a surface by the surface's own shape and placement relative to a light source. Intrinsic shadow contours provide useful information about the direction of the light source and the qualitative shape of the underlying surface. We analyze the invariants relating surface shape and light source direction to the shapes and singularities of intrinsic shadow contours. The results suggest that intrinsic shadows can be used to directly infer illuminant tilt, qualitative global surface structure, and, at intersections with surface creases, the concavity/convexity of a surface. We show that the results obtained for point sources of light generalize in a straightforward way to extended light sources, under the assumption that light sources are convex. PMID- 9392901 TI - Does the photon-diffusion coefficient depend on absorption? AB - We investigate the controversy over the precise form of the photon diffusion coefficient and suggest that it is largely independent of absorption, i.e., Do = v/3mu(s)'. After presentation of the general theoretical arguments underlying this assertion, Monte Carlo simulations are performed and explicitly reveal that the absorption independent diffusion coefficient gives better agreement with theory than the traditionally accepted photon diffusion coefficient, D(mu)a = v/3(mu(s) + mu(a)). The importance of resolving this controversy for the proper characterization of the material optical properties is discussed. PMID- 9392902 TI - Use of four mirrors to rotate linear polarization but preserve input-output collinearity. II. AB - We report on the design, construction, and testing of a four-mirror reflective polarization rotator, proposed by Smith and Koch [J. Opt. Soc. Am. A 13, 2102 (1996)], that rotates by an angle phi the input linear polarization while preserving the input-output beam collinearity. We correct errors in the previous work that led to an incorrect design for a phi = pi/2 rotator. This type of pure rotator is simple and inexpensive, and it is a direct application of the concept of the nonadiabatic geometric phase to polarization rotation. We also present measurements of the polarization rotation for the case of three metallic mirrors with antiparallel input and output beams, a test of geometric phase in polarization optics not done before. PMID- 9392903 TI - Decorrelation of L- and M-cone signals. AB - For a large sample of broadband lights reflected from natural and man-made objects, the correlation between L- and M-cone absorptions was found to be 0.99. The correlation between L + M and L - M signals was 0.21. The early recombination of cone signals in the visual system thus leads to a substantial decorrelation. PMID- 9392904 TI - Secular rates of twinning in Asia: recent observations and review of literature. AB - OBJECTIVE: To study the rates of twinning in Asian countries using the most recent data available. METHOD: A Medline search was performed and all primary and secondary references obtained. RESULTS: Data was obtained from 5 Asian countries from reports published between 1983 and 1993. Total twinning rates varied. CONCLUSION: Twinning among Asians is much more variable than had been reported previously. Further documentation in this area is indicated. PMID- 9392905 TI - Ectopic pregnancy in uterosacral ligament. AB - Two cases of ectopic pregnancy in the uterosacral ligament are presented. The cases are reported not only because of its rarity but also to arouse the thought of primary aetiology. With the increasing incidence of ectopic pregnancy due to assisted reproduction, gynaecologists are posed with the diagnostic challenge. The golden rule of management in extra-tubal pregnancy is to maintain high index of suspicion. PMID- 9392907 TI - Hydatid cyst of Morgagni: any impact on fertility? AB - On laparoscopy in 3 patients with primary infertility, the only pathology found was hydatid cysts of Morgagni that were excised. In one patient with monolateral hydatid cyst, pregnancy failed to be achieved. Despite prior failure of repeated trials of ovulation induction and intrauterine insemination in the other 2 patients, a spontaneous pregnancy was achieved within 2-3 months following laparoscopic extirpation of hydatid cysts of Morgagni. The hydatid cysts were bilateral in one case; and monolateral (in relation to the only present tube with a unicornuate uterus) in the other case. It is concluded that hydatids of Morgagni, as a single pelvic pathology, might hinder fertility. Laparoscopic extirpation of these cysts would improve ovum pick-up and enhance fertility. PMID- 9392906 TI - A study of tubo-ovarian and ovarian abscesses, with a focus on cases with endometrioma. AB - OBJECTIVE: To determine the incidence and causes of endometrioma-associated tubo ovarian abscesses (TOAs) and ovarian abscesses. METHODS: The medical records of 6,557 gynecologic inpatients were reviewed, and the data were analyzed using the median test and chi 2 test. RESULTS: The incidence of TOAs was 2.3% (5/218) in patients with endometrioma, and 0.2% (11/6,339) in patients without endometrioma (p = 0.0001). Among the TOA cases (n = 16), no significant differences in age, parity, history of pelvic surgery, or isolated organisms were observed between the subgroups with (n = 5) and without endometrioma (n = 11). There were only 2 cases of ovarian abscess, and both were associated with endometrioma. The causes of the abscesses in the 7 cases with endometrioma were contamination during surgery (1 case), contamination during a transvaginal endometrioma aspiration (1 case), and ascending infection (1 case), and unknown in 4 cases. CONCLUSIONS: The presence of endometrioma is a risk factor for the development of a TOA or an ovarian abscess. PMID- 9392908 TI - Diffuse cystic change of a term placenta with a normal newborn. AB - We recently encountered a case of term placenta with a diffuse cystic lesion of the villi. A 19-year-old primipara at 36 weeks of gestation underwent cesarean section due to breech presentation with premature rupture of the membranes; she delivered a mature male baby of 2,502 g with an Apgar score of 9/9. The placenta was 940 g in weight and 29 x 20 x 3 cm in size, and macroscopically had multiple cystic lesions (3-8 mm in diameter) that resembled hydatidiform moles. However, histopathological examination revealed that the severe hydropic change was localized in the stem villi but not remarkable in the terminal chorionic villi. Moreover, abnormal proliferation of the trophoblast was not observed. However, the hypertrophic change was observed in the vascular wall of stem villi, in which hyperplasia of smooth muscle-like cells was present. The urinary hCG levels at 1 month and 2 months after delivery were less than 50 IU/l. These findings indicate that the multiple cystic lesions of the placenta in this case are essentially different from those of a trophoblastic disease, and that the diffuse cystic lesion of the villi might have been secondary to changes in the local circulation. PMID- 9392909 TI - Spontaneous resolution of a postcesarean arteriovenous fistula of the uterine cervix: the usefulness of transvaginal color Doppler scanning. AB - We encountered in a woman at the 28th day after cesarean delivery the transient appearance of a uterine cervical mass with a prominent blood flow, that was revealed to be an acquired arteriovenous fistula. The size and blood flow of the mass were monitored using transvaginal color Doppler scanning and magnetic resonance imaging (MRI). Based on these findings, the arteriovenous fistula was managed conservatively without any adverse complications. PMID- 9392910 TI - Dengue hemorrhagic fever during pregnancy: antepartum, intrapartum and postpartum management. AB - Dengue hemorrhagic fever is a common tropical disease in Thailand that nowadays has an increasing incidence during adulthood. We managed three cases of dengue hemorrhagic fever during pregnancy which developed during the antepartum, intrapartum and postpartum periods. We diagnosed dengue hemorrhagic fever during pregnancy with clinical pictures of fever, hemoconcentration and thrombocytopenia with serological proof in all cases. All cases were treated conservatively except for the second one, in which cesarean delivery was inevitable due to severe preeclampsia with unfavorable cervix. All patients and their babies were in good condition before discharge. With increasing incidence during adulthood, more cases of dengue hemorrhagic fever in pregnancy can be found. Conservative treatment should be conducted in all cases. We believe that this is the first case report of intrapartum dengue hemorrhagic fever during pregnancy. PMID- 9392911 TI - Spontaneous rupture of the liver in an uncomplicated pregnancy. AB - Spontaneous liver rupture has been reported during pregnancy secondary to severe pregnancy-induced hypertension, eclampsia or syndrome of hemolysis, elevated liver enzymes, and low platelet count (HELLP). Here we report a case diagnosed using CO2 intra-arterial digital subtraction angiography in an uncomplicated pregnancy. A 33-year-old Chinese woman at 39 weeks' gestation underwent a second caesarean delivery. Her pregnancy course had been uneventful. In the immediate postpartum period, she developed clinical signs of hepatic infarction or hematoma. After a blood transfusion and the use of vasoactive agents, her hemodynamic condition became stable. A hepatic angiography was performed before an emergent laparotomy. CO2 intra-arterial digital subtraction angiography revealed a small extravasation that was not found by the conventional method using an ionized medium. This disease should be considered when there occur pain in the upper part of the abdomen and signs of hemorrhagic shock, even in the case of an uncomplicated pregnancy. PMID- 9392912 TI - Prenatal diagnosis of Duchenne muscular dystrophy in the Japanese population by fluorescent CA repeat polymorphisms analysis. AB - OBJECTIVE: To investigate the efficacy of CA repeat analysis using fluorescence labeled primers in the prenatal diagnosis of Duchenne muscular dystrophy in the Japanese population. METHODS: Allelic frequencies of polymorphic loci in the Duchenne muscular dystrophy gene were ascertained, and the polymorphic information content (PIC) was calculated. CA repeat analysis of 21 Japanese families with Duchenne muscular dystrophy was then performed. RESULTS: The STR 49 locus had the highest PIC, followed in decreasing order by the loci of STR 44, STR 45, STR 50, DYS III, DYS II, DYS I, and 3' CA. The diagnostic applicability increased to 0.999 when the PICs of all 8 loci were combined. When the highest PIC from each of the 5' end, exons near the 3' end and the 3' end were combined, the diagnostic applicability increased to 0.988. Of the 7 males examined prenatally, 1 was diagnosed as normal, and 6 were affected, while of the 9 females examined, 5 were diagnosed as carriers, and 4 as non-carriers. CONCLUSION: CA repeat analysis using fluorescence-labeled primers is useful in the prenatal diagnosis of Duchenne muscular dystrophy in the Japanese population. PMID- 9392913 TI - The immunotherapy during in vitro fertilization and embryo transfer cycles in infertile patients with endometriosis. AB - OBJECTIVE: To investigate if the immunotherapy with corticosteroids would improve the pregnancy rate in infertile patients with endometriosis who undergo in vitro fertilization and embryo transfer (IVF-ET). METHODS: Forty-two infertile patients with endometriosis plus tubal factor and 87 pure tubal infertility patients who underwent IVF-ET in our unit were allocated randomly to the corticosteroid treatment group and the control group. RESULTS: The prevalence of autoantibodies (antinuclear antibody, lupus anticoagulant, anticardiolipin antibody, rheumatoid factor) was elevated significantly in patients with endometriosis plus tubal factor compared with pure tubal infertility patients (38.1% vs 2.3%). Twenty-one patients with endometriosis plus tubal factor underwent 54 cycles of IVF-ET, receiving corticosteroids. Forty-three patients with pure tubal factor underwent 81 cycles of IVF-ET, receiving corticosteroids. Twenty-one patients with endometriosis plus tubal factor who underwent 57 cycles of IVF-ET and 44 patients with pure tubal factor who underwent 84 cycles of IVF-ET served as controls, not receiving corticosteroids. In patients with endometriosis plus tubal factor, there was a significantly higher clinical pregnancy rate per cycle in the treatment group, with 42.6% (23/54) compared with 22.8% (13/57) in the control group but no differences between 2 groups in spontaneous abortion rate (21.7% vs 15.4%) and multiple pregnancy rate (17.4% vs 15.4%). In patients with pure tubal infertility, there were no significant differences between the treatment group and control group in clinical pregnancy rate (40.7% vs 34.5%), spontaneous abortion rate (12.1% vs 10.3%) or multiple pregnancy rate (18.2% vs 10.3%). In the endometriosis plus tubal infertility group with autoantibodies, the clinical pregnancy rate per cycle was significantly higher in the treatment group at 40.9% compared with 14.8% in the control group. In endometriosis plus tubal infertility group without autoantibodies, there was no significant difference between 2 groups with respect to the clinical pregnancy rate per cycle (43.8% vs 30.0%). CONCLUSIONS: This study suggests that immunotherapy with corticosteroids could improve the clinical pregnancy rate in endometriosis patients undergoing IVF-ET and may be more effective in patients with positive autoantibodies. PMID- 9392914 TI - A study of gamma-aminobutyric acid (GABA) in amniotic fluid. AB - OBJECTIVE: The purpose of the study was to evaluate the role of gamma aminobutyric acid (GABA), an inhibitory neurotransmitter, in amniotic fluid (AF) during fetal distress, because it has been reported that several neurotransmitters, e.g. norepinephrine, are affected by GABA. METHODS: AF was obtained during elective cesarean section (CS, n = 11) and cesarean section due to fetal distress without labor pain (FD, n = 7). Maternal and umbilical-cord blood, as well as the first urine of the neonates, also were collected. GABA, norepinephrine (NE), and epinephrine (EP) concentrations were measured using HPLC. RESULTS: The GABA concentration was higher in the AF than in either maternal or fetal circulation, or in the first urine of neonates. The GABA concentrations in the AF and in the first urine of neonates were significantly higher in the FD group than in the CS group (p < 0.05). Furthermore, significant positive correlations were observed between the NE and GABA concentrations and between the EP and GABA concentrations in the AF. GABA was produced in a time dependent manner in cultured amnion cells. CONCLUSION: The highest concentration of GABA was found in the AF. The GABA in the AF appeared to be derived from both the amniotic membrane and the fetal urine. The increase in the GABA concentration in cases of fetal distress might be partially derived from the fetus via fetal urine. The positive correlations between the concentrations of GABA and those of NE and EP in the AF, suggest that GABA, NE, and EP might play important roles during fetal distress. PMID- 9392915 TI - Effects of fibrin glue on postsurgical adhesions after uterine or ovarian surgery in rabbits. AB - OBJECTIVE: To evaluate the prophylaxis with fibrin glue on postoperative adhesions following uterine or ovarian surgery in rabbits. STUDY DESIGN: The uterine horns of 10 rabbits were exteriorized and electrocauterized by monopolar coagulation. Fibrin glue was applied to one uterine horn and the other was left untreated as a control (Experiment 1). Multiple ovulation was induced in 12 rabbits with hCG, and wedge resection was performed on both ovaries of each animal. One ovary was treated with fibrin glue and the other was untreated as a control (Experiment 2). Second-look laparotomy was performed to assess adhesion formation 2 weeks after the initial operation. RESULTS: The use of fibrin glue resulted in significant decrease in postoperative adhesions compared with the control side, in the uterine horns and the ovaries (p < 0.02 and p < 0.01, respectively). CONCLUSION: Fibrin glue was useful in preventing postoperative adhesions of the reproductive organs in rabbits. PMID- 9392916 TI - Integrin adhesion molecules in the endometrial glandular epithelium in patients with endometriosis or adenomyosis. AB - OBJECTIVE: To assess the role of beta 1-integrin and E-cadherin molecules in the eutopic glandular epithelium in patients with endometriosis or adenomyosis. STUDY DESIGN: Twenty-four patients with endometriosis, and 22 patients with adenomyosis diagnosed histologically were selected as subjects. The controls consisted of 29 fertile women. Eutopic endometria were obtained by curettage or immediately after the operation. The samples were immunohistochemically examined for the expression of very late activation antigen-2 (VLA-2), VLA-3, VLA-4, VLA-5, VLA-6, and E cadherin. RESULTS: The expression of each VLA molecule and E-cadherin except VLA 4, VLA-5, and VLA-6 was significantly increased throughout the menstrual cycle in endometria in both the endometriosis and adenomyosis groups. In contrast, the expression of VLA-4 in the adenomyosis group was significantly reduced in the secretory phase. CONCLUSION: Altered expression of beta 1-integrins and E cadherin was observed throughout the menstrual cycle in patients with endometriosis and adenomyosis, suggesting the defective microenvironment of the endometrium. PMID- 9392917 TI - False-positive urine drug screen: beware the poppy seed bagel. PMID- 9392918 TI - Postural hypotension from topical glyceryl trinitrate ointment for anal pain. PMID- 9392919 TI - Assessment and knowledge in palliative care in second year family medicine residents. AB - Inadequate physician knowledge, particularly in areas of pain assessment and use of analgesics, has been identified as a major factor contributing to poor pain management in cancer patients. In most medical schools, teaching in Palliative Care at both the undergraduate and postgraduate levels is limited or nonexistent. Baseline knowledge and changes in knowledge in areas relevant to Palliative Care were assessed by the use of 2 16-question examinations (Exams A and B) in 78 second-year Family Medicine Residents from the University of Alberta Family Medicine Residency Program. The residents participated in a two-week rotation on the Acute Palliative Care Unit at the Edmonton General Hospital or Grey Nuns Community Health Centre between September 1991 and February 1996. The residents were randomly assigned on their first day (Time 1) to complete either Exam A or B and were subsequently crossed over on their final day of the rotation to complete the alternate Exam (Time 2). Six domains were represented in the Exams as follows: pain assessment, opioid use, adjuvant medications, delirium, urinary catheterization, and hydration. Improvements were noted in the mean percentage results in Time 2 compared with Time 1 for Exams A, B, and A and B combined. Mean global percentage results were 53 +/- 15 versus 73 +/- 13 (p < 0.001) at Times 1 and 2, respectively. There were significant improvements for domains in Time 2 compared to Time 1 (p < or = 0.05) for combined A and B Exam except for urinary catheterization. Despite these documented improvements in scores, serious deficiencies were identified particularly in the areas of pain assessment and opioid use, namely opioid sude effects and issues involving dependence, addiction, and tolerance. Examinations, such as the two used in this study, can be a useful aid in assessing physician knowledge in addition to structuring teaching in Palliative Care. Examination content will require updating as knowledge in Palliative Care evolves. PMID- 9392920 TI - A role model program to promote institutional changes for management of acute and cancer pain. AB - This report describes an 18-month project to make acute and cancer pain management an institutional priority in Southeastern Wisconsin health-care facilities. Facility-based teams, each of which included a nurse in a leadership position, were recruited to participate in a project based on the Cancer Pain Role Model Program. The project was conducted in three stages: (a) a 1-day conference focusing on basic pain management issues and clinical standards, (b) a preceptorship at the Medical College of Wisconsin, and (c) a follow-up conference focusing on institutional change. Participants completed an Action Plan, outlining activities aimed at changing practice in their facility. Participants from 17 of the 32 participating facilities partially or completely met their Action Plan goals. Lack of ongoing facility commitment, staff turnover and facility closures were cited as reasons for failure to meet goals. Nurses in key positions, provided with strong institutional commitment and given suitable educational training and nurturing, are ideally suited to help facilitate changes in institutional pain practices. PMID- 9392922 TI - Backlash in the treatment of cancer pain: use of opioid analgesics in a Finnish general hospital in 1987, 1991, and 1994. AB - Finland belongs to the group of countries in which the consumption of strong opioids is low. This seems to reflect the general quality of cancer pain treatment. During the last 10 years, many efforts have been made to improve the treatment of cancer pain in Finland. To assess one parameter of change, the present study compared the quantity of opioid and nonopioid analgesics used in the treatment of terminal cancer pain in a Finnish general hospital in 1987, 1991, and 1994. Specifically, the records of all patients who died of cancer in Kymenlaakso Central Hospital (KCH) in 1991 and in 1994 and during the last 6 months of 1987 were reviewed to acquire information about the use of analgesic medication. The total proportion of cancer patients receiving analgesic medication on a regular basis was 39% in 1987, 63% in 1991, and 52% in 1994. The mean daily dose of strong opioids changed from 24 mg in 1987 to 58 mg in 1991, and to 43 mg in 1994. These data suggest a possible backlash in prescribing practices during recent years. In spite of various efforts to improve the treatment of cancer pain, the medical records demonstrate a decline in prescribing of the drugs needed for this treatment. PMID- 9392921 TI - Graduating medical students' competencies and educational experiences in palliative care. AB - Palliative care involves an interdisciplinary approach to patient care and specific clinical skills. Little prior research on palliative care education has involved medical students, and the few reported studies focus mainly on student attitudes. This study describes a needs assessment of senior medical students based on a newly developed competency-based palliative care curriculum. Prior to graduation, 102 senior students were mailed an anonymous survey with four parts: a self-assessment of attitudes, knowledge, and skills; adequacy of instruction; exposure to specific clinical experiences; and demographic information. The response rate was 47%. While attitudinal goals were strongly endorsed by students, they were less confident with regards to knowledge and skills. Ratings varied across the five content areas of the curriculum. The results suggest a need for educational efforts more focused on specific clinical competencies as well as systematic evaluation of student competencies. PMID- 9392923 TI - Assessment of satisfaction with treatment for chronic pain. AB - The purpose of this study was to develop an instrument to assess satisfaction with treatment of chronic pain, evaluate the reliability and validity of this instrument, and then examine predictors and consequences of satisfaction. The Pain Service Satisfaction Test (PSST) is the result of this effort. Fifty adult patients receiving services for chronic pain in a university pain clinic completed the PSST as part of a survey mailed to their homes. Findings supporting the validity of the PSST included significant positive correlations with a general measure of treatment satisfaction, patient ratings of global treatment satisfaction and effects of treatment, and physician ratings of patient satisfaction with treatment. Regression analyses of predictors of satisfaction highlighted significant contributions of confidence and trust in the provider, pain reduction, and waiting in the clinic. These predictors together accounted for 60% of satisfaction with treatment. Treatment satisfaction was negatively correlated with depression, reported number of physicians consulted, and number of physician visits for pain in the past 12 months; and there was a trend toward a negative correlation with disability. Results of the present study support the importance of satisfaction with treatment as a predictor and possible determinant of later health, function, and service utilization. PMID- 9392924 TI - Nurses' perceptions of factors influencing the use of a pain program. AB - Factors that, according to nurse participants, influenced the application of what was learned in a pain program were explored by means of qualitative interviews. Participants indicated that the correspondence between the program and their personal view on pain management, attitudes toward the program and innovations in general, self-efficacy perceptions, and (un)familiarity and taboos with respect to program items influenced what they put into practice. In addition, participants indicated that interactions with colleagues, nursing managers, patients, and physicians affected their application of the program. Furthermore, organizational factors, such as limited time and lack of formal program implementation, were mentioned as influential. PMID- 9392925 TI - Intractable nausea and vomiting due to gastrointestinal mucosal metastases relieved by tetrahydrocannabinol (dronabinol). AB - Four years following resection of a Clark's level IV malignant melanoma, a 50 year-old man developed widespred metastatic disease involving the liver, bones, brain, gastrointestinal mucosa, and lungs. One week after whole brain radiation therapy, he was admitted to the hospital for nausea, vomiting, and pain. He was treated with several antiemetic drugs, but it was not until dronabinol was added that the nausea and vomiting stopped. Dronabinol was an effective antiemetic used in combination with prochlorperazine in nausea and vomiting unresponsive to conventional antiemetics. PMID- 9392926 TI - STDs surprisingly prevalent--diagnosis should be pursued aggressively. PMID- 9392927 TI - Sexually transmitted diseases: a review. PMID- 9392928 TI - The approach to treatment of invasive pneumococcal disease in the 1990s. AB - Streptococcus pneumoniae is the most common cause of pediatric invasive infections and an important cause of morbidity and mortality. In the past, S. pneumoniae responded universally to penicillin until nonsusceptible isolates were first noted in the 1960s. Before 1990, penicillin-nonsusceptible isolates remained a minor component of all reported isolates. Since that time, 20-30% of isolates in many centers in the United States and up to 50% of isolates in some other countries are penicillin-nonsusceptible. Of greater concern has been the development of isolates which are nonsusceptible to more than one antimicrobial agent. This review presents data on pediatric invasive pneumococcal disease in Arkansas and outlines the new treatment recommendations which have been developed in response to these problems. Streptococcus pneumoniae is an important pathogen worldwide and is considered the most common etiology of bacterial sinusitis, otitis media, pneumonia, meningitis and bacteremia. Before 1990, 95-96% of pneumococcal isolates were susceptible to penicillin. The first report of penicillin-nonsusceptible S. pneumoniae was made by Hansman and Bullen in 1967, who identified the strain in the sputum of a patient with hypogammaglobulinemia. Soon thereafter, penicillin-nonsusceptible pneumococci were reported in New Guinea and Australia as well. Over the last several years, the incidence of penicillin-nonsusceptible isolates has greatly increased. Of particular concern is the concomitant increase in the number of organisms that are nonsusceptible to more than one antimicrobial agent. Due to the development of such isolates, clinicians are having to approach patients with invasive disease due to pneumococci more cautiously. In an attempt to clarify confusion with terminology, the Centers for Disease Control and Prevention (CDC) have recommended the same nomenclature be used to classify resistance for all organisms: nonsusceptible organisms are those with an MIC (minimal inhibitory concentration) greater than or equal to that defined for the intermediate category of resistance and the term resistant should be reserved for those organisms with an MIC greater than or equal to that defined for the resistant category. Therefore, resistant isolates are a subgroup of the nonsusceptible isolates. PMID- 9392929 TI - Hindsight--20/20. PMID- 9392930 TI - Acute aortic dissection. PMID- 9392931 TI - Rabies in Arkansas. PMID- 9392932 TI - Results of the Kentucky high school football knee injury survey. AB - The study attempted to: 1) determine knee injury and reinjury incidence of Kentucky high school football players; 2) relate results to initial care provider, treatment following initial physician exam, time lost from injury, injury type, player position, and team size; and 3) assess coaches opinions and practices about lateral prophylactic knee brace (LPKB) usage and effectiveness. A post season, mail-in coaches' survey (50.2% return, 101/201) collected these data. Returned surveys represented 4690 players with average team size (x +/- SD) of 43 +/- 13. Two hundred fifty seven reported knee injuries yielded .055 knee injuries/player with .04 knee injuries/player being "new" and .015 knee injuries/player recurring (27% of reported knee injuries) during the season. Games accounted for 56.4% (56/101) of reported knee injuries. Coaches generally believed that LPKB usage prevented knee injuries (56.4%, 56/101) and allowed LPKB usage (92.1%, 93/101), however only 8.3% (8/101) required their wear (interior linemen 50%, linebackers 25%, entire team (25%). Interior linemen had the greatest number of knee injuries, followed by offensive backs and linebackers. Most knee injuries (81%, 208/257) were out 3-6 weeks or less, 64% (164/257) involved sprains or contusions, 38% (97/257) were treated surgically (alone or with rehabilitation) and 36% (92/257) were treated solely with rehabilitation. Total knee injury and reinjury incidence were under-estimated compared to existing reports. Improved injury recording methods, and post-symposia coaches evaluation are recommended. PMID- 9392933 TI - Prolymphocytic leukemia: diagnosis and treatment. A case report. AB - A patient with B-cell prolymphocytic leukemia (PLL) who has had a prolonged survival is presented. The patient was diagnosed incidentally while asymptomatic, but later developed progressive disease. He was refractory to alkylating agents and fludarabine, but responded to treatment with cyclophosphamide, doxorubicin, vincristine, and prednisone. This patient's prolonged survival may be due partly to his diagnosis at an indolent phase, possibly representing the early phase of the natural course of the disease. Diagnosis, clinical course, and treatment options for PLL are discussed. PMID- 9392934 TI - Interprofessional code. Kentucky Medical Association and Kentucky Bar Association. PMID- 9392935 TI - The coming plague. PMID- 9392936 TI - Audits, investigations, and serious trouble. PMID- 9392938 TI - Direction of medical care--one physician's view. PMID- 9392937 TI - Committee to investigate changing trends in medicine. Report on fraud and abuse laws. PMID- 9392939 TI - Toxic Pfiesteria and human health. AB - Toxic activity of a Pfiesteria-like organism occurred for much of 1997 in the waters of the lower Pocomoke River on Maryland's Eastern Shore. Maryland's experience with these toxic blooms of dinoflagellates, current knowledge of their potential human health effects, and the actions taken by state government agencies in response to a potential public health threat are reviewed. A medical diagnostic team commissioned by the Department of Health and Mental Hygiene evaluated a group of persons with intense exposures to lesioned fish or the waters from which they came and/or prominent symptoms following exposure to affected waters or lesioned fish. The principal findings of the team included consistent complaints of memory problems, acute burning of the skin following direct contact with water, and respiratory irritation. Findings on examination were limited to neurocognitive deficits in short-term memory and learning difficulties. Physicians and citizens are asked to continue to report, through their local health departments, illnesses thought to be related to exposure to lesioned fish or the waters from which they are taken. Persons with questions or wishing to report finding lesioned fish should call the state Pfiesteria hotline at 1-888-584-3110. PMID- 9392940 TI - Diagnosis of Pfiesteria-human illness syndrome. AB - The first case reports of human illness caused by exposure to Pfiesteria piscicida toxin(s) acquired outside of a laboratory are reported. Though Pfiesteria, a toxin-forming dinoflagellate, is responsible for killing billions of fish in estuaries in North Carolina, its role in human illness has remained controversial, in part due to lack of identification of the toxin. A recent fish kill in the rivers of the lower Eastern Shore has permitted careful investigation and identification of a distinct clinical syndrome resulting from exposure to the Pfiesteria toxin--Pfiesteria human illness syndrome (PHIS). Patients have memory losses, cognitive impairments, headaches, skin rashes, abdominal pain, secretory diarrhea, conjunctival irritation, and bronchospasm. Not all patients have all elements of the syndrome. PMID- 9392941 TI - A 74-year-old man with persistent fevers. AB - An elderly man with a history of extensive world travel presents with a chronic illness and fevers. The febrile illness has been present for eight years, and no diagnosis has been made despite extensive evaluation and testing. The differential diagnosis of this unusual case of fever of unknown origin is discussed. PMID- 9392942 TI - Ankle sprains: evaluation, treatment, rehabilitation. AB - Ankle sprains are a common, costly, and potentially disabling problem. The proper history and physical examination will determine the need for radiological evaluation and treatment. Complications of ankle trauma like osteochondral fractures, peroneal tendon injuries, fracture of the os trigonum, synovial impingement, tarsal tunnel syndrome, Achilles tendon inflammation or rupture, and nerve injury are reviewed. The treatment of ankle sprains is based on the severity of the injury. Treatment begins with rest, ice, compression, and elevation. Casting and orthotics may be needed to facilitate healing. Primary rehabilitation, functional rehabilitation, and performance testing and the assessment of efficacy for each of these modalities are critical parts of proper treatment for ankle sprains. PMID- 9392943 TI - Heroic medicine, bloodletting, and the sad fate of George Washington. AB - Bloodletting was an established medical treatment for more than two millennia, yet its greatest impact in the United States is undoubtedly the role it played in the treatment of President George Washington. The theoretical justification for bloodletting is provided, followed by a detailed description of the treatment Washington received, reflecting the role played by heroic medicine in the American president's demise. The arguments of bloodletting's critics emerge as well founded; indeed, Washington and many others might have suffered less had heroic medicine not been applied. PMID- 9392944 TI - Interviews with women medical society leaders. PMID- 9392945 TI - New recommendations for screening infants and children for tuberculosis. PMID- 9392946 TI - Reader reflects on caring for the terminally ill. PMID- 9392947 TI - Domestic abuse: a social ill in need of a cure. PMID- 9392948 TI - Curing cancer takes more than medicine. PMID- 9392949 TI - Dr. Walter Kempner and the Rice Diet program: another reason why Durham is the City of Medicine. PMID- 9392950 TI - Next generation telemedicine. The future is now. PMID- 9392951 TI - Oral contraceptive pills. Prevention of ovarian cancer and other benefits. PMID- 9392952 TI - Postmenopausal hormone replacement therapy. Information for effective patient counseling. PMID- 9392953 TI - For patients: hormone replacement therapy. PMID- 9392954 TI - Investor-owned or not-for-profit health care. A conundrum for communities. PMID- 9392956 TI - Environmental tobacco smoke: the smoke that's around you. PMID- 9392955 TI - Some thoughts on keeping our noble profession professional and noble. PMID- 9392957 TI - Gastroesophageal reflux disease. Pill, blade, or laparoscope? PMID- 9392958 TI - Psychosocial factors and coronary disease. A national multicenter clinical trial (ENRICHD) with a North Carolina focus. AB - In addition to traditional risk factors (cigarette smoking, high blood pressure, and elevated cholesterol) psychosocial factors (depression, social isolation, and low socioeconomic status) have an adverse impact on prognosis of patients with CAD. Several studies of psychosocial and behavioral treatments provide encouraging evidence for the clinical efficacy of psychosocial interventions in CAD patients. A new, multicenter clinical trial now underway (see sidebar) will evaluate the impact of psychosocial interventions (compared to usual care) on all cause mortality and nonfatal MI in post-MI patients with depression or perceived low levels of social support or both. PMID- 9392959 TI - "Children are not supposed to die." Combined pediatric and radiation oncology grand rounds addresses severe illness and death. PMID- 9392960 TI - How to open a free medical clinic. PMID- 9392961 TI - The health and health care utilization of people in Buncombe County. PMID- 9392962 TI - "Boldly they rode ... into the mouth of hell". Pennyroyal oil toxicity. PMID- 9392963 TI - Automobile hood ornament as penetrating missile. PMID- 9392965 TI - Acheson revisited: public health medicine ten years after the Acheson Report. AB - The issue of communicable disease control and the role of public health medicine is once more of considerable concern, particularly in the light of recent outbreaks and NHS reorganisations. Ten years ago, following similar concerns, the Acheson Report highlighted several issues with striking parallels to the present day. These included uncertainty over the future and role of public health medicine following repeated Health Service reorganisations. The Report recommended the expansion of the specialty in both communicable disease control and public health medicine in general, and a more clearly defined role for public health medicine. Successive health authority mergers and the specialty's inclusion as part of management costs have meant that the Report's findings have yet to be fully implemented. In fact, the establishment of public health medicine consultants (CsPHM) has fallen by a quarter since 1992. A further review of the public health function is required if the specialty is not to fragment, and if able doctors are not to be deterred from entering the specialty. Such a review should include the removal of the specialty from management costs, and the clarification & standardisation of the roles of the Director of Public Health (DPH), CPHM and other members of the multi-disciplinary public health team, as well as identifying possible organisational locations for the public health function. PMID- 9392964 TI - Health professionals--do we have enough? PMID- 9392966 TI - Organ transplanting in Japan: the debate begins. AB - OBJECTIVES: Japan is currently considering changing its long-standing policy of banning most organ transplants. This paper reviews the current state of organ transplantation in Japan and presents results from a recently conducted survey regarding attitudes toward the removal of organs from brain-dead donors, and potential methods of allocating those organs in a fair manner. METHODS: Survey data were collected by the Research Project Team on Network Systems for Organ Transplants funded by the Japanese Ministry of Health and Welfare. The sample consisted of 1093 randomly selected citizens. Predictors of attitudes supporting organ transplantation were analyzed using logistic regression. RESULTS: Although many Japanese people support organ transplantation, few are willing to donate their organs. General knowledge of transplantation was the best predictor of support for such a program and willingness to donate organs. In addition, younger respondents and male respondents were more likely to support programs and donate organs. Implications of these results are discussed. PMID- 9392967 TI - A serological survey of measles vaccine in a rural region of Eskisehir in Turkey. AB - We designed this longitudinal study as a response to measles outbreaks which occur occasionally in Eskisehir in Turkey to investigate the incidence of primary and secondary vaccine failure. The investigation was conducted over two periods (December 1993 to October 1994 and April 1995 to October 1995). Two study groups were involved, infants aged 9-11 months and children aged 18 months to 9 y. During December 1993 to October 1994 prevaccination sera was collected from 35 infants aged 9-11 months and tested to determine if maternally derived antibodies were present. The infants were vaccinated and subsequently the 31 infants who could be traced were retested 30-40 d later to determine their response to vaccination. The following was done to determine whether seropositivity rates alter over time. The second group, a randomised sample of 117 children aged between 18 months to 9 y was chosen; all of whom had been previously vaccinated and who had no history of measles. Sera was taken and tested during December 1993 to October 1994 in order to determine whether seropositivity rates varied with time. During April 1995 to October 1995 out of all the children in both groups 123 children were retested. All sera samples were studied by an enzyme-lined immunosorbent assay (ELISA). Out of the 35 infants in group 1 only four (11.4%) had maternal antibody against measles on initial testing. Out of 31 infants followed up 30-40 d after vaccination (61.3%) were found to be seropositive for measles. In the second group, of the 117 previously vaccinated children ninety three (71.5%) had measles IgG antibody. Seropositivity rates did not show significant difference with time after vaccination. There was no association between first and second screening seropositivity rates. We conclude that the presence of maternal antibody reduces the success of vaccination. These results suggest the vaccination policy in Turkey should be re-examined with a view to revision. PMID- 9392969 TI - Maternal serum screening for Down's syndrome: a survey of midwives' views. AB - The lack of consensus regarding the implementation of maternal serum screening, has led to a widespread variation in practice. The importance of the role of midwives within the service has been recognised. All maternity units in the Northern region now have a designated 'co-ordinator' in an attempt to improve service delivery, professional liaison and training. This study was designed to obtain midwives' views about maternal serum screening in principle and to assess whether any changes had occurred since the introduction of co-ordinators. Semi structured, postal questionnaires were sent to all midwives in one health authority for them to complete and return. Within this authority, one maternity unit offered universal screening whereas the other maintained a selective policy. Responses were obtained from 90 out of 133 (67.7%). There was almost unanimous support for the principle of screening 86 out of 90 (95.5%) and most midwives considered the offer of screening should be an NHS service, independent of age 78 out of 90 (86.7%). Half of the respondents 46 out of 90 (51.2%) reported that the introduction of a co-ordinator had been successful in improving staff education but requests for further training and updating were made by 69 out of 90 (76.6%) despite having had this organized training input: although those midwives who were regularly involved with screening made significantly fewer requests 27 out of 45 (60%). These findings confirmed our previous recommendation that ongoing responsibility for such provision would be required. The results of the study provided a useful contribution towards the review of screening policy undertaken by the health authority, as well as evidence upon which to base further development of the role of the co-ordinators in their support of midwives. PMID- 9392968 TI - Use of contraception in women who present for termination of pregnancy in inner London. AB - OBJECTIVES: To describe the contraceptive usage of women undergoing termination of pregnancy in order to identify problems with contraception, and therefore suggest ways in which contraceptive services can be improved. DESIGN: Prospective study of attenders for NHS termination of pregnancy over a three month period. SETTING: Community based assessment clinics for NHS termination of pregnancy in inner London. SUBJECTS: Two hundred and sixty-nine women asking for assessment for NHS termination of pregnancy. MAIN OUTCOME MEASURES: Source of contraception, method used around time of conception, and problems experienced. RESULTS: Respondents tell into three groups: those using contraception around the time they became pregnant; those who had ceased to use contraception; and those that had never used contraception. The method of contraception used by the majority of the first group was the condom and the main source of the method was the chemist shop. The second group had most commonly used oral contraceptives in the past and had ceased use in many cases as a result of side effects. The majority of the third group did not speak English and had limited knowledge of methods of contraception. CONCLUSIONS: High usage of chemists means women avoid service providers who could offer help and advice. Women were prepared to put themselves at risk of unwanted pregnancy rather than return for further help and the lack of knowledge about emergency birth control was of some concern. The needs of black and ethnic minority women requires detailed work to improve access and acceptability of contraceptive services. PMID- 9392970 TI - How good is the evidence relating to the frequency of childhood sexual abuse and the impact such abuse has on the lives of adult survivors? AB - Concerns relating to the methodological rigour of studies examining the frequency and effects of childhood sexual abuse (CSA) have comforted those who dismiss the results as scaremongering. This review highlights the difficulties to be overcome in interpreting the epidemiological data currently available, and the lack of consensus regarding the true frequency of CSA and the impact such abuse has on the lives of adult survivors. However, it concludes that the rates of reporting and disclosing a history of CSA are increasing, and that the health service needs to recognise and respond to this changing clinical picture. PMID- 9392971 TI - Somatic morbidity in schizophrenia--a case control study. AB - Several studies have stated that rates of premature mortality of schizophrenic patients are increased. Morbidity, however, is less often examined. In the present study we have compared the number of hospitalizations due to different somatic diseases in 775 schizophrenic patients with their sex- and age-matched controls. The total number of patients hospitalized due to somatic diseases was significantly greater among the schizophrenics than among the controls. In the former group, 523 out of 775 and in the latter 373 out of 775 had been admitted at least once during a 15 y period. If the schizophrenic patients who also had been diagnosed as substance abusers at least once were excluded from the analysis there was still a significant difference between the remaining patients and their controls. The patients exhibited an over-morbidity in almost all diagnostic groups, the most prominent excess morbidity appeared in the groups of injuries and symptoms, signs and ill-defined conditions, when compared with the controls. Even if there are confounding factors which should be taken into consideration when interpreting morbidity data, the pattern of excess morbidity among schizophrenic patients found in this study is so definite that it cannot be considered merely coincidental. PMID- 9392972 TI - Public views on community involvement in local health services in South Africa. AB - An analysis of the information collected in a nation-wide health survey shows that the South African public favours the involvement of communities in local public health services. There are variations in the support for community involvement in four aspects of health services examined, namely decision on opening times of clinics, determination of patient-provider relationship, recruitment of staff and determination of service charges. Multivariate analysis indicates that the level of support for community involvement is significantly low for Whites relative to other races, lower for rural residents than for city dwellers and high for people with a very good health status. Further focused research is required for improved understanding of the problems and policy options in community involvement in public health programmes. Since 1994 the new government has consistently expressed a strong commitment to involve communities in the design and implementation of health policies and programmes. However, considerable amount of uncertainty remains on how to translate such commitment into practical action at the local level. PMID- 9392973 TI - Approaches to community concerns: applied public health. AB - Public health workers are challenged to address community health concerns believed to be related to environmental exposures. The challenge is heightened when there are multiple potential exposures and numerous health concerns. In a community in Washington State, we employed different approaches depending on the specificity of diagnosis and the relation of the disease to environmental exposures. For diseases with specific diagnoses and questionable associations with environmental exposures, we began by determining whether rates of disease were higher than expected. Different approaches were needed for three different health concerns. Major limitations were estimating community population counts and obtaining comparison rates from published literature. Despite limitations, epidemiologic data developed at a relatively low cost were useful in assisting the community in understanding its health status. PMID- 9392974 TI - Supply of in-patient medical services for elderly people and geographical variation in medical admissions in a health district in England. AB - Geographical variation in the utilisation of in-patient medical services for elderly people in a health district in England was examined in relation to supply of in-patient geriatric medical care and indicators of need. An ecological study design based on electoral wards was used. The health district had a resident population of 67,919 aged 65 y or more and was divided into three localities, each with a different supply of in-patient medical services for elderly people. Locality A had a traditional model of geriatric medical care, Locality B an integrated model and Locality C an age-related model. Localities A and C also had a high provision of general practice hospital beds. The main outcome measure was the age and sex standardised hospital admission ratio for people aged 65 y or more admitted under geriatric medicine, general medicine or general practice in April 1991-March 1992. There were 8829 admissions in 1991/2, 48% in general medicine, 40% in geriatric medicine and 12% in general practice, giving an overall unadjusted admission rate of 130 per 1000 population aged 65 y or more for the three specialties combined. Locality A had the highest, and Locality B the lowest, unadjusted admission rate for the three specialties combined. This rate remained highest in Locality A if second and subsequent admissions in the same period were excluded. Lengths of stay in geriatrics were longest in this locality but lengths of stay for the three specialties combined were similar in the three localities. Multiple regression was used to examine the effect of three indicators of need, the Jarman score, standardised mortality ratio and prevalence of limiting long-term illness, on standardised admission ratios at the electoral ward level. Jarman score had a significant independent association with the standardised admission ratio but adjustment for this factor did not alter the ranking of the three localities, with the standardised admission ratio remaining highest in Locality A. Subject to the limitations of the study, the results suggest that factors related to the supply of in-patient medical services may be associated with geographical variation in medical admissions for elderly people. PMID- 9392975 TI - Introducing managed care in Switzerland: impact on use of health services. AB - The objectives of this study were to assess changes in the self-reported use of health care services after gatekeeping by general practitioners and a global budget were introduced in the health insurance plan for students at the University of Geneva, Switzerland, in October 1992. A random sample of 336 members of the University plan answered questions about their use of health care services during the year before (1992) and the year after (1993) the introduction of managed care. Similar data were collected among a random sample of 300 members of a comparison plan. All participants were 18-44 y old in 1992, spoke French and lived in Geneva. The proportion of insurees who visited specialists decreased by 10% in the University plan between 1992 and 1993 and remained unchanged in the comparison group. The proportion of insurees who visited general practitioners increased by 12% in the University plan and remained unchanged in the comparison group. No effects on the total number of health care visits, on hospitalisations or on use of medications were detected. The introduction of gatekeeping and of a global budget managed by physicians was associated with a transfer of patient visits from specialists to general practitioners. PMID- 9392976 TI - Cellular phones and traffic accidents. AB - Cellular phone use in motor vehicles is becoming an increasing world-wide phenomenon. Using data obtained from traffic accidents reported between 1992 and 1995 in the state of Oklahoma, USA, this study examined statistical rate-ratios of accident characteristics between drivers with or without cellular phones. Rates were calculated between cellular phone involvement and reported accident causes, types of collision, driver actions immediately prior to the accident, location of the accident, the extent of fatalities, and age and gender of drivers. Results indicated a significant increased rate among drivers with cellular phones for inattention, unsafe speed, driving on wrong side of road, striking a fixed object, overturning their vehicle, swerving prior to the accident, and running off the roadway. People with phones stood an increased risk of being killed in an accident over persons without phones. Males with phones had a significantly higher rate than females for many of accident characteristics mentioned above. Rate-ratios of some accident characteristics and fatalities increased as age increased, with the exception of drivers under age 20 yrs, who had the highest fatality rate. Limitations of the study and possible prevention alternatives are discussed. PMID- 9392977 TI - A medical cause of death validation study of adult aboriginal deaths in the Northern Territory of Australia in 1992. AB - Judged on the criterion of equity, premature adult Aboriginal mortality is the most serious public health problem faced in Australia today. There have been a number of published epidemiological studies that have analysed Aboriginal cause of death data, but this is the first study to formally validate such data. The study sample included all adult Aboriginal people who lived and died in the Northern Territory in 1992, excluding residents of the Alice Springs region. The appropriateness of underlying cause of death codes was assessed by a single reviewer in light of death certificates, medical records, postmortem records and interviews with key health professional informants. Data were collected on 220 deaths. 8% (17 out of 220) of deaths were classified erroneously at the ICD-9 chapter level. Errors in death certification accounted for 64% (11 out of 17) of the chapter errors and diagnostic and coding errors for 18% (3 out of 17) each. The overall impact on mortality statistics was less severe because some cross chapter classification errors cancelled each other out. Misclassification errors aggregated mainly in chapter VII (circulatory diseases) of the ICD-9 classification which was overcounted by 3.2%, and chapter VIII (respiratory diseases) which was overcounted by 1.3%. Before correction for misclassification error, circulatory diseases were judged to cause the highest proportion of deaths, whereas after correction, respiratory diseases accounted for the highest proportion. Despite this, the overall quality of the medical cause of death statistics was of a sufficiently good standard from a public health perspective to broadly inform health policy. Future attempts to improve the validity of medical cause of death statistics for Australian Aboriginal people should focus on the education of medical practitioners about the purpose and process of death certification. PMID- 9392978 TI - Expanded programme on immunization (EPI). Progress towards global measles control and elimination. PMID- 9392979 TI - AIDS-related disseminated histoplasmosis in San Francisco, California. AB - The published reports of patients with the acquired immunodeficiency syndrome (AIDS) with disseminated histoplasmosis come mostly from institutions located in endemic areas for histoplasmosis, where disease is thought to occur by either primary infection or reactivation. The characteristics of reactivation disease are not well delineated. We describe the clinical features of reactivation disseminated histoplasmosis in 46 residents of San Francisco, California, with AIDS who did not report recent travel to an area endemic for histoplasmosis. Patients presented with illness lasting days to months, manifested most frequently by fever, chills, sweats, cough or dyspnea, gastrointestinal complaints, malaise, and weight loss. Physical examination and imaging studies were notable for hepatosplenomegaly, lymphadenopathy, or abnormal pulmonary findings in more than half of patients. Laboratory studies revealed a high rate of cytopenia, elevated serum lactate dehydrogenase levels, abnormal liver function test values, respiratory alkalosis with hypoxemia, and a median CD4 lymphocyte count of 36 x 10(9) per liter. The clinical presentation of reactivation disseminated histoplasmosis in patients with AIDS living in San Francisco is similar to that of disseminated histoplasmosis reported in patients with AIDS living in endemic areas. Reactivation disseminated histoplasmosis should be considered in any AIDS patient with a low CD4 lymphocyte count, a febrile illness, and a history of travel or residence in an endemic area. PMID- 9392980 TI - Intramuscular high-dose triamcinolone acetonide in the treatment of severe chronic asthma. AB - We describe our experience with administering intramuscular triamcinolone acetonide to 22 steroid-dependent patients with asthma. These patients represent the minority of those with asthma whose disease is characterized by frequent emergency department visits, hospital admissions, and long-term dependency on oral corticosteroid therapy. The participants were randomly assigned to 2 treatment groups, one group receiving 120 mg of intramuscular triamcinolone acetonide, the second receiving 360 mg as a series of three 120-mg daily doses. We determined relative efficacy by comparing peak expiratory flow rates and incidents of emergency department visits, hospital admissions, and ventilatory failure of the study and during the 12 months before enrollment. Peak expiratory flow rates improved significantly in both groups. The mean (+/- standard deviation [SD]) monthly percentage of predicted peak expiratory flow on the study was 88.6 +/- 3.7% and 91.2 +/- 3.9% compared with 63 +/- 15.1% and 64 +/- 14.5% at entry in patients receiving 120 and 360 mg, respectively (P < 0.02). Patients receiving 120 mg required 8 hospital stays and 8 emergency department visits compared with 27 hospital stays and 72 emergency department visits in the previous year (P < 0.05). Patients receiving 360 mg required 5 hospital stays and 5 emergency department visits compared with 33 hospital stays and 34 emergency department visits in the previous year (P < 0.05). The average monthly interval (+/- SD) between exacerbations was 2.7 +/- 2.3 and 7.8 +/- 3.5 for patients receiving 120 mg and 360 mg, respectively. A total of 25 intubations was required in the previous year and only 1 during the study. The incidence of cushingoid facies, weight gain, and hypertension was reduced in both groups (P < 0.05). Total steroid use was reduced in both groups (P < 0.02). A dose of 360 mg produced a longer exacerbation-free period than 120 mg (P < 0.02). PMID- 9392981 TI - Geographic distribution, supply, and need for generalist physicians in Alaska. AB - This study provides the first comprehensive description of Alaska's geographic distribution of generalist physicians relative to population. All 443 generalist care physicians (family, general, general internal medicine, and pediatric) or their office managers were questioned about their specialties, ZIP codes, employers, populations served, and hours spent per week offering direct patient care. The results indicated a 30% overall shortage of generalist physicians for the state, representing roughly 141 full-time-equivalent generalists relative to national practice patterns and trends of health maintenance organizations. Of 17 primary health care areas, including the Anchorage area, 15 showed a need for additional generalist physicians. Most areas had a 20 to 40% shortage. Concerns about transportation and financial barriers to access to care, especially in remote regions, were raised. Other needs emphasized included knowledge of contributions of midlevel health care professionals, Alaska Native versus non Native care, efforts to train and retain physicians in Alaska, and the need for longitudinal tracking of practice patterns. PMID- 9392982 TI - Closer to home (or home alone?) The British Columbia long-term care system in transition. AB - Finding ways to organize and deliver long-term care that provides for quality of life at an affordable price is of increasing importance as the population ages, family size decreases, and women enter the workforce. For the past 2 decades, British Columbia has provided a model system that has apparently avoided disruptive conflicts. Although formal users' complaints are rare, this study- based on focus groups and interviews with users, their families, and advocates- identified problems users encountered toward resolving concerns about the structure, process, and outcome of long-term care. We present these findings in the context of British Columbia's current devolution from provincial to regional control that aims to save costs and keep disabled elderly persons in the community. British Columbia may be continuing to lead the way in meeting the needs of its burgeoning elderly population for long-term care. Study findings have implications for the development of US long-term care policy by pointing to the value of obtaining users' views of long-term care to identify both obvious and more subtle trouble spots. PMID- 9392983 TI - Recommendations for immunizations in HIV-infected individuals. PMID- 9392984 TI - Drug reactions in HIV/AIDS. PMID- 9392985 TI - Latex allergy--the latest insights. PMID- 9392986 TI - Antileukotrienes--1997 and beyond. PMID- 9392987 TI - Combination therapy for HIV disease. PMID- 9392988 TI - Advances in allergic rhinitis pharmacotherapy. PMID- 9392989 TI - Treatment of chronic urticaria. PMID- 9392990 TI - Inhaled corticosteroids for asthma therapy. PMID- 9392991 TI - Intravenous immune globulin in neuromuscular disorders. PMID- 9392992 TI - Paralytic shellfish poisoning in Kodiak, Alaska. PMID- 9392993 TI - Resolution of propylthiouracil-induced hepatic failure after treatment of thyrotoxicosis. PMID- 9392994 TI - Trimethoprim-sulfamethoxazole associated with hyperkalemia. PMID- 9392995 TI - Treatment of chronic severe asthma. PMID- 9392998 TI - Schistosomiasis: the cleanup continues. AB - During the past 40 years immense progress has been made in the control of schistosomiasis in China. The authors consider how the 100 million people in the country who are still at risk from the disease can be protected. PMID- 9392997 TI - Ethics, equity and renewal of WHO's health-for-all strategy. AB - The renewal of WHO's strategy for health for all raises questions about human and societal values that have to be fully taken into account. Discussions to date have highlighted the principle of equity in the context of ethics and human rights. PMID- 9392999 TI - Cardiovascular disease stoppable in developing countries? AB - The author discusses factors with a bearing on cardiovascular disease which are shared by many developing countries, and outlines the difficulties that have to be overcome if significant prevention is to be achieved. PMID- 9393000 TI - Assessing risk factors for chronic diseases. AB - A pilot survey of men aged 35 and over in Buenos Aires indicated that many had one or more risk factors for chronic diseases. A low response rate hampered the investigation: on average it proved necessary to visit several homes in order to obtain one interview. Furthermore, at the cost of US$ 10 incurred per interview, large prospective investigations would be precluded in most developing countries, but case-control studies assessing tobacco use and other risk factors retrospectively would be a good alternative. PMID- 9393001 TI - Health sector reform: priorities and packages. AB - Measures of health sector reform have been adopted in Chile, Colombia and Mexico. The extent to which they promote equitable access to appropriate health services is considered below. PMID- 9393002 TI - Health information systems--making them work. AB - A health services model based on different concentration levels between the centre and the periphery, each with particular resources, responsibilities and management functions, provides a framework on which health information systems can be built or rebuilt so as to accelerate progress towards the health-for-all goals. PMID- 9393004 TI - Reducing maternal mortality in St Petersburg. AB - Following the entry of St Petersburg into Europe's Healthy Cities Project in 1991 it was decided that the highest priority should be given to reducing the city's maternal mortality ratio, then standing at approximately 70 deaths per 100,000 live births. Preventing deaths from unsafe, illegal abortion became the main focus of attention. The use of modern contraceptive methods was promoted, information was disseminated to improve the utilization of family planning services, special outreach services for teenagers were established, and providers were given opportunities for education and training. The maternal mortality ratio and the abortion rate have now declined and contraceptive use appears to be increasing. These achievements are attributable in large measure to the commitment of a broad spectrum of St Petersburg society as well as to outside support. PMID- 9393005 TI - Training in district health management. PMID- 9393003 TI - Sociocultural aspects of haemorrhage in pregnancy. AB - The knowledge, attitudes and practices of rural women in southern Nigeria are at least as important as the availability of modern obstetric care in the fight against haemorrhage in pregnancy. Community-based interventions taking this into account are necessary if the considerable mortality associated with the condition is to be significantly reduced. PMID- 9393006 TI - Medical negligence: a return to trust. PMID- 9393007 TI - What food for the heart? PMID- 9393008 TI - Medical aid for refugee camps. PMID- 9393009 TI - Management of Bartholin's abscess. PMID- 9393010 TI - Female health workers boost primary care. AB - Women residing in villages in three districts of Pakistan were recruited, trained to deliver primary care and mobilize their communities for health, assigned to limited catchment areas, provided with supervisory and managerial support, and remunerated. Their comprehensive activities substantially reduced infant, child and maternal mortality within a year and generated positive perceptions of family planning in the communities. The programme was cost-effective and appeared suitable as a model for reforming the organization and provision of health care services. PMID- 9393012 TI - AIDS prevention with local implementors--overcoming obstacles. PMID- 9393011 TI - Partnership for primary care. AB - A project for improving primary health care in an underserved rural area of Osun State, Nigeria, involved the creation of a partnership between the local government, the community and a medical college. Joint administrative and technical committees were established, and community mobilization was fostered. The evidence so far indicates that partnership designs can accelerate the development of primary health care in an affordable manner. PMID- 9393013 TI - Improving the performance of health centres in district health systems. Report of a WHO Study Group. AB - Health centres are in the front line of health care provision. Located within communities and usually easily accessible, health centres are in a key position not only to contribute to improved health for the individual but also to foster the development of the community as a whole. This report of a WHO Study Group looks at how health centres function in different parts of the world, at the range of services they provide, and how they can be made both more efficient and more effective. The report covers political and legal issues, management, resources and personnel, finance, and the provision of high-quality health care that matches clients' needs. It describes how health centres can design their activities to suit the health profile and priorities of the communities they serve. The report acknowledges past drawbacks and future challenges. The health centre's role is a changing one but, as this report shows, it is a role of growing importance for the future as individuals and communities take on increased responsibility for their own health and well-being. PMID- 9393014 TI - Why complementary medicine is so popular. PMID- 9393015 TI - Abortion law matures. PMID- 9393016 TI - Back to nature. PMID- 9393017 TI - A shot in the arm for public health. PMID- 9393018 TI - Unfinished business. PMID- 9393019 TI - The nurses helped the family do the right thing. PMID- 9393020 TI - The mental health service user movement. PMID- 9393021 TI - Clinical supervision: a hornet's nest. PMID- 9393023 TI - Addiction. The vice of fame. PMID- 9393022 TI - Clinical supervision: a hornet's nest? ... or honey pot? PMID- 9393025 TI - A time to die. PMID- 9393026 TI - The electronic campus. PMID- 9393024 TI - Practical procedures for nurses. 4.2. Respiration technique and observation--2. PMID- 9393027 TI - Lung cancer specialist and respiratory registrar. Interview by Alison Whyte. PMID- 9393028 TI - The management of fluid balance. AB - This is the sixth part in the series Care is Critical, which looks at the more complex interventions nurses now have to deal with on general wards and in the community. This article looks at the clinical assessment and management of fluid balance disturbances. PMID- 9393029 TI - Alginate dressings in wound care management. PMID- 9393030 TI - Self-esteem and stress in mental health nurses. PMID- 9393031 TI - Lean times for skin care. PMID- 9393032 TI - Dermatology: the forgotten subject? PMID- 9393033 TI - Know how. Wet wraps in atopic eczema. AB - Wet wraps play an important part in the management of children with eczema that does not respond to first-line treatment. They are used in hospitals and in the community, and provide great relief and comfort, but the procedure requires considerable input from the health team until the parents are confident of their technique. PMID- 9393034 TI - Take three experts. Interview by Adam Legge. PMID- 9393035 TI - The assessment of patients with malignant fungating wounds--a holistic approach: Part 1. PMID- 9393036 TI - Advances in laser technology make this an exciting time in plastic surgical nursing. PMID- 9393037 TI - Laser physics and physiology. AB - Laser light begins when an excited and unstable electron moves from its unstable state back to a more stable state producing energy in the form of a photon. Laser light is coherent which means that the light waves move in phase together in space and time. Laser light is monochromatic which means it is comprised of only one color or wavelength. Laser light is also collimated which means it is perfectly parallel and travels in a single direction with very little divergence. Medical lasers fall in the infrared and visible as well as ultraviolet portion of the electromagnetic spectrum and are available at different wavelengths. The wavelength of each laser partially determines the effect it will have on tissue. A specific wavelength or color can be used to selectively target a specific tissue such as hemoglobin, water, or melanin. Heat is produced by the laser, destroying the targeted tissues. PMID- 9393038 TI - Laser blepharoplasty. AB - Laser blepharoplasty was introduced in 1984 by Sterling Baker and although there are few disadvantages, several advantages as compared to traditional blepharoplasty have been assessed. These advantages include an improved scar, a shorter operating time, and a more rapid recovery time in most patients. Methods include laser resurfacing of the periorbital area or full face with excision of the upper eyelid skin and fat, as well as excision of the lower eyelid fat through a transconjunctival incision alleviating the need for a lower eyelid scar. In some cases, an upper eyelid excision may not be necessary following resurfacing of this area. Excellent results have been achieved in over 150 cases with comparisons of scarring in scalpel versus laser incisions. PMID- 9393039 TI - Laser resurfacing of the face. AB - Pulsed mode carbon dioxide laser allows precise ablation of fine facial tissue while minimizing thermal damage to the skin. Changes in the structure of dermal collagen may account for the overall tightening effect observed. Preoperative preparation of the skin is important to prevent postoperative pigment changes. Prophylactic antivirals are used to reduce the risk of Herpes infection. The carbon dioxide laser produces pain when applied to the skin and various anesthetic techniques may be used. After lasing, the face can be dressed with occlusive dressings or left open. Sunscreens are required after reepithelization. PMID- 9393040 TI - Complications of laser resurfacing. AB - The laser demonstrates significant benefits over traditional resurfacing and incisional techniques. Serious complications of laser surgery are easily avoidable provided practitioners and support staff receive proper education and training. The following article describes the more common complications of laser resurfacing followed by a brief discussion of avoidance techniques. PMID- 9393041 TI - Hair removal with the ruby laser (694 nm). AB - Hair removal with the ruby laser is one of the newest uses of laser technology. The laser seeks melanin in the hair shaft, and melanin content is highest during the growth phase of the hair follicle. Nursing care focuses on preparing the patient for the procedure, maintaining a safe operative environment, and teaching the patient skin care after the laser therapy. PMID- 9393042 TI - Confidentiality--An analysis of the issue. AB - Today, in the high-tech, fast-paced health care arena in which professionals practice, confidential information and the right to privacy in one's personal life have become very basic concerns of society. Among the general public, a genuine interest exists about potential invasions of privacy and the undermining of confidentiality. Members of society are more educated concerning rights as the result of multimedia and education presented by health care professionals. New challenges for the professional attending to the health needs of society will be encountered as the complexity of health care increases (Pohlman, 1988). One must remember that the most intensely private and personal moments of an individual's life are revealed in hospital settings. Health care professionals have a duty at all times to diligently protect the confidence of each patient unless specific criteria is met so that disclosure is warranted. Dilemmas encountered often are complex and require a certain degree of expertise for resolution to occur. If ethical theory and principles are used in combination with some of the specific guidelines apropos to resolving confidentiality issues, creative solutions can be devised. Professionals, by the very nature of their experience and education, possess the talent to achieve this goal. Consequently, when guidelines for confidential treatment of information are followed, public trust of professionals will be ensured. Once trust is established, a significant step will have been taken toward the solution of problems of confidentiality in modern day health care. PMID- 9393043 TI - Research critique of pulsed dye laser treatment of port-wine stains: a patient questionnaire. PMID- 9393044 TI - Laser safety guidelines for plastic surgical nurses. AORN. PMID- 9393045 TI - To lease or not to lease. PMID- 9393046 TI - Laser surgery: sample patient education materials for plastic surgical nurses. PMID- 9393047 TI - Use of a semipermeable dressing (Biobrane) following laser resurfacing of the face. AB - Biobrane provides a suitable, reliable method of postoperative wound coverage for the laser resurfaced patient. Pain and drainage are minimized. PMID- 9393048 TI - Expanding practice and obtaining consent. AB - The Scope of Professional Practice liberates nurses, midwives and health visitors from previous role constraints, while keeping the focus on the patient. Some of the adjustments to the scope of practice require the individual practitioner to obtain explicit verbal or written consent. PMID- 9393049 TI - Performing peripheral intravenous cannulation. AB - Nurses need to be aware of their level of accountability when performing i.v. cannulation. Nurse cannulation should be incorporated into practice as part of holistic patient care. When introducing nurse cannulation into a department, a commitment is required from the individual nurse and the organisation. i.v. cannulation is a skilled technique, best performed as a staged process. PMID- 9393050 TI - Pressure-relief mattresses and patient comfort. AB - The ideal pressure-relieving support system is comfortable, relieves pressure and prevents tissue damage. In order to ensure patient compliance with the choice of mattress, patient comfort and quality of sleep should be among the most important factors involved in the decision-making process. PMID- 9393051 TI - Parkinson's disease. PMID- 9393052 TI - Parenteral nutrition. AB - Patients with a non-functioning gastrointestinal tract, those who require bowel rest or cannot receive enteral nutrition will be considered for parenteral feeding. This Update looks at indications, venous access, administration and complications. PMID- 9393053 TI - Creating an education model for cardiac patients. AB - Patient education is important in reducing the risk of recurrence of myocardial infarction. Education should be specific to patients' individual needs. Patients need motivating to achieve the required behaviour changes. A multidisciplinary, multifactorial approach to education is most effective. PMID- 9393054 TI - Nursing and the law. PMID- 9393055 TI - Professional negligence. AB - The rules on which the law of negligence is based derive from judgements and judicial statements contained in medical and non-medical cases. The principles documented in these cases apply equally to nurses and it is therefore important for nurses to be aware of them. PMID- 9393056 TI - Liability in negligence. AB - The nurse has two forms of liability, first for the action he or she took and, second, for the harm that resulted. Nurses can take several simple measures to protect themselves against liability for negligence. PMID- 9393057 TI - Sterilising solutions for heat-sensitive instruments. AB - Disinfectants should be used only to decontaminate heat-sensitive medical equipment. A COSHH risk assessment is required to ensure the safe use of disinfectants. Equipment and processing manufacturers should be consulted before using any of the alternatives to glutaraldehyde. PMID- 9393058 TI - Peri-operative nursing. PMID- 9393059 TI - [ICN's representatives meet in Vancouver]. PMID- 9393060 TI - [ICN Congress]. PMID- 9393061 TI - [Goodness in nurse's heart matters. What is goodness in nursing practice?]. AB - Manifestation of goodness in nursing has been researched very little contrary to good care. This article is based on a Master's thesis the aim of which is to describe goodness in nursing as it is experienced by a nurse. The research is qualitative and based on the phenomenological philosophy of science in which the truth is understood through an individual experience. The research method is developed by Paterson and Zderad. There were 15 nurses who participated in the research and the following results were derived from their essays. Life experience and prevailing values of society influence on the nurse's values which are reflected in nursing practice. The nurse's sensitivity, maturity, flexibility, hardiness and ability to grow combined with technical skills facilitate goodness. Goodness is an open relationship with another people. In this constantly living relationship closeness, listening by heart and willingness to bring good to the other people exist simultaneously. PMID- 9393062 TI - [Depression in humans--experience with being helped]. PMID- 9393063 TI - [Cipramil project--recruitment wrong from the start]. PMID- 9393064 TI - [Falck problem--3,880 ambulances only arrive after 20 minutes]. PMID- 9393065 TI - [Narcotics Council--county's methadone treatment is inadequate]. PMID- 9393066 TI - [Psychiatry--in a hospital bed outside the hospital. Interview by Grethe Kjaergaard]. PMID- 9393067 TI - [Sexuality discontinued specifically in education]. PMID- 9393068 TI - [ADDH children suffer from distorted hearing]. PMID- 9393069 TI - [Supervisory nurses are indispensable]. PMID- 9393070 TI - [Quality development--passport to project pitfalls. Interview by Susanne Block Kjeldsen]. PMID- 9393071 TI - [Psychiatry--case manager for Helle]. PMID- 9393072 TI - [Patient information--ageism or care?]. PMID- 9393073 TI - [Emergency medicine--new alarm system can save human lives]. PMID- 9393074 TI - [Psychiatry--greatest advantage with open day facilities. Interview by Grethe Kjaergaard]. PMID- 9393075 TI - [Psychiatry--here in our house. Interview by Grethe Kjaergaard]. PMID- 9393076 TI - [Misuse of syringes--clean syringes should limit the harm. Interview by Jesper Berg]. PMID- 9393077 TI - [Fetal monitoring as a service available to everyone]. PMID- 9393078 TI - [Romania--poverty forces children into an institutionalized life]. PMID- 9393079 TI - [Basic nursing care]. PMID- 9393080 TI - [Latex allergy--a current threat]. PMID- 9393081 TI - [Health care: observation of infants]. PMID- 9393083 TI - [Rogaland psychiatric hospital takes violence seriously]. PMID- 9393082 TI - [Relatives: lack of confidence in the system]. PMID- 9393084 TI - [Intensive care--mostly for active 30-year-olds]. PMID- 9393085 TI - [Viewpoint from the housekeeper. Interview by Marit Fonn]. PMID- 9393086 TI - [Part-time work--every third one wants longer working hours]. PMID- 9393087 TI - [Equality--male applicants get preference. Interview by Kari Anne Aase]. PMID- 9393088 TI - [Closeup Anders Vege, leader of the Nominating Committee in Norwegian Nurses' Association. Start of the hunt. Interview by Marit Fonn]. PMID- 9393089 TI - [My workplace: emergency admissions unit, Haukeland Hospital, Bergen. Where time is both friend and enemy. Interview by Erik Dale]. PMID- 9393090 TI - [From earlier times. A national conference in equal rights spirit]. PMID- 9393091 TI - [The aged--love is ageless. Interview by Erik Dale]. PMID- 9393092 TI - [Clinical nursing--taking care of eyes. Interview by Kjell Arne Bakke]. PMID- 9393093 TI - [Russia--the goal is independence. Interview by Marit Fonn]. PMID- 9393095 TI - [Professional Ethics Council: ethics, staffing and budget]. PMID- 9393094 TI - [Femidom started in fight against AIDS]. PMID- 9393096 TI - [Elderly immigrants--fear old age in Norway]. PMID- 9393097 TI - [Drug dependence is not always an addiction problem]. PMID- 9393098 TI - [Sexual assault--examination should not be another kind of assault]. PMID- 9393099 TI - [Minister Borst on the cutting nurse]. PMID- 9393100 TI - [The shadow disciplinary tribunal meets]. PMID- 9393101 TI - [The new nursing scientist. Interview by Tonny van de Pasch]. PMID- 9393102 TI - [Flexible child care important for health care sector]. PMID- 9393103 TI - ['Nurses: remain at the bedside']. PMID- 9393104 TI - [Palliative Center Ghent straightens out the last winding road]. PMID- 9393105 TI - [Terminal dehydration]. PMID- 9393106 TI - [LCVV starts training program for young management talent (leadership for experts in nursing and health care)]. PMID- 9393107 TI - [From the diary of a district nurse]. PMID- 9393108 TI - [Advanced nursing practice]. PMID- 9393109 TI - [Universities develop Master's education advanced nursing practice. Master or doctorandus?]. PMID- 9393110 TI - [Sense and nonsense of protective isolation]. PMID- 9393111 TI - [Council BIG ceases to exist (professions in individual health care). Inspired across the finish line]. PMID- 9393112 TI - [Flexible child care]. PMID- 9393113 TI - [Malnutrition in the hospital]. PMID- 9393114 TI - [The soul of caring]. PMID- 9393115 TI - [Diagnosis in nursing: the patient and his family]. PMID- 9393116 TI - [Ground-breaking nursing]. PMID- 9393117 TI - [AZR (Academic Hospital Rotterdam) determines restricted procedures. Interview by Corina de Feijter]. PMID- 9393118 TI - [Nursing care and quality assurance. Interview by Tonny van de Pasch]. PMID- 9393119 TI - [Moving forward in AIDS care. 9th annual ANAC Conference in Chicago (Association of Nurses in AIDS Care)]. PMID- 9393120 TI - [Nursing and self-regulation. Interview by Petra Nijdam]. PMID- 9393121 TI - [Norwegian research]. PMID- 9393122 TI - [Clinical supervision--experience of 10 nurses of a 2-year process-oriented supervision]. AB - Many patients in swedish hospital care today are older, more physically and mentally ill and in need of much care. Despite this there is a strong urge for higher cost-effectiveness and shorter hospital stays. Both these trends are often reported to be important sources of stress and burn-out among nurses. One way of reducing such effects among nurses could be clinical supervision. The present study is a result of one two-year long process-oriented supervision programme for nurses, led by the authors. The method in this study consisted of semi-structured interviews of 10 nurses joining the programme. The main questions in the interviews concerned; a) what knowledge they had acquired through the programme; b) if the programme had contributed to their understanding of others' feelings and reactions and; c) whether the programme had influenced their cooperation with others. The main conclusion from the interviews was that the programme had given them more courage and a more pronounced experience of support from colleagues. It was also found that the programme had improved their sense of professionalism and their self-image. These results point to the value of offering nurses possibilities for supervision, giving opportunities for both personal and professional development and thus preventing burn-outs. PMID- 9393123 TI - [We do not create values, but uphold them--a contribution to nursing's value theory]. AB - This article argues how the nature of nursing is anchored in a realist conception of moral value. This moral ontology claims that phenomena like suffering, pain and human distress have a distinct moral character that is independent of individual human perception and empathic responsiveness. This position of moral realism is evident in the ethics of KE. Logstrup, Kari Martinsen and Nel Noddings. However, these influential theories are in need of a philosophical theory of moral agency which establish a role for moral sensitivity. Moreover, and trying to bridge the gap between analytical moral philosophy and fenomenologist analysis in ethics, the article's final part illuminates how the encounter with the moral realities of immediate human suffering inhabits a significant normative claim. This normative claim of compassion and mercy restricts an impartial, justice-based morality and has significant implications on nurse's perceptions of prioritizing dilemmas. PMID- 9393124 TI - [How do nurses attend to their educational function and how do patients experience this? A field study in an orthopedic department]. AB - Patient teaching is an essential component of patient care. Even if patient education has been given much attention the last year--both in research and textbooks for health professionals, some patients still complain about too little information and support when they are in the hospital. The purpose of this study was to find out how nurses fulfil their teaching responsibility in practice. Why, what and how do they teach patients? How do they document their teaching? What do patients think is the most important in relation to patient teaching? The study was performed as a field study in an orthopaedic ward. Participation, open observation and semistructured interviews were used as methods. Notes from observation, interviews and written nurse documentation are the data in the study. The results showed that the nurses were teaching the patients while they were doing some practical things at the same time. Nurses did not plan the teaching, and in hectic periods less information was given. Most of the teaching was information about facts. Patients were told little about what they actually would feel or sense (sensory information)--even if this is what several patients said they wanted to know about. The patients wanted to meet fewer people and they often felt that the nurses signalized business so they hesitated to ask them questions. A standard teaching program was not used. Most of the teaching was given orally, but written materials and a video were also used sometimes. The nurses did not document their teaching. PMID- 9393125 TI - Urinary incontinence among a 65-year old Swedish population: medical history and psychosocial consequences. AB - Urinary incontinence (UI) is a disability caused by an impairment, which can lead to a handicap of importance for nursing care. This means that UI is not only a practical-medical concern but also a socio-economic problem. The purpose of the study was to determine the prevalence of UI among 65 year-olds in a Swedish Health Care District and to compare gender differences concerning medical history and psychosocial consequences. In a Primary Health Care District, a questionnaire pertaining to UI was mailed to all women and men 65 years of age (N = 458). A total of 91% (n = 419) was sufficient for data analysis, which was performed by descriptive and inferential statistics. It was found that 28% (n = 61) of the women and 9% (n = 21) of the men were afflicted with UI. Women reported significantly more urge incontinence (p < .05) as well as stress incontinence (p < .05). Information from the health service about UI had been given to 46% (n = 28) of the women and 33% (n = 7) of the men. The strongest reason reported, both in women (42%, n = 26) and men (40%, n = 8), for not seeking help from the health service was that UI was a normal condition for people of their age. Most of the women had to urinate at least twice per night (42%) compared to once per night (44%) for the men. It is important to establish a UI clinic at every main Primary Health Care Centre which builds on nursing care and whose aim is to inform the general public that UI is a common problem, that it leads to psychosocial consequences, and that the health service can offer active rehabilitation interventions. PMID- 9393126 TI - [Letting family participate in care. A study on relatives' experience with a palliative geriatric care department]. AB - An evaluation was made of relatives' experiences of the care of the patient and the treatment of themselves at a geriatric ward where elderly patients dying of cancer were nursed. A questionnaire was sent to 100 relatives after they had agreed on telephone to participate. 86 relatives answered the questions. The results shows that the majority of the relatives were satisfied with the care of the patient and the treatment of themselves as relatives. Most of them thought that they had been able to influence the care of their next-to-kin. However, elderly ladies and younger men were uncertain whether they had had the possibility to influence the care. Most of the relatives had been given the opportunity to talk enough to the doctor, the nurses and the social-worker. However; they required a more active approach from the staff in the ward. They also desired more information about the patient's disease, prognosis and medical treatment. The findings of this study suggest that relatives need information repeated at several occasions in order to understand the message. They also need support and invitation to get the courage to ask questions and to participate in the care of their dying next-of-kin. We need deepen our knowledge of what kind of information the relatives demands and what information needs to be repeated. Evaluations of routines to make contact easier between patients, their relatives and hospital staff should be done continuously. PMID- 9393127 TI - [Tell me what you are like--a study of mentorship conferences in health and social work graduate studies]. AB - The purpose of this study was to investigate educational systems with regard to mentoring practice in two Schools of Health and a School of Social Work. Student mentor interaction, notably mentor receptiveness, was studied through storytelling and discourse analysis from a practical/theoretical perspective. The study revealed that clinical mentors competencies are insufficient. As would be expected, clinical mentors, however, are superior to college teachers in practical mentoring situations. The study is based on observations of six different mentoring situations in three different groups/contexts. PMID- 9393128 TI - [Health status of the elderly over 67 years old in a community, and their need of nursing care. A survey with descriptive analysis. Health status in the light of life style]. AB - The purpose of this descriptive study was to explore the problems of community dwelling persons 67 years and older, in meeting their self-care requisites, and the factors in living arrangements associated with these problems. Community nurses in both administration and practice can use the results in planning curative and preventive interventions for all the elderly or for groups of elderly according to living arrangements. These groups were those living alone, living with a spouse or living with other family members. A standardized questionnaire revised from Dr. Agnes Bjorns interview guide used in Denmark, was mailed to 893 persons 67 years and older residing in their own homes in an Eastern Norwegian community. To analyse the data, Dorothea Orems categories of self-care requisites was used. The method was usefull to find the health status among the elderly. Many elderly were satisfied with their lifestyle even if they had health problems. Memory problems was the biggest area of problems, 59% mentioned this. An average of 30% were dissatisfied with each of 10 specified bodily functions. The data totally affirmed greater problems among those elderly living alone or with a family member, than among those elderly living with a spouse. The literature says little or nothing about basic need problems among elderly living with a family member. PMID- 9393129 TI - [A long and rocky road in a Swedish community. Interview by Anders Olsson]. PMID- 9393130 TI - [Many fine words but little action]. PMID- 9393131 TI - [The profession needs us, immigrants. Interview by Maria Ejd]. PMID- 9393132 TI - [Communities learn ever more about health care--thanks to our nurses. Interview by Kaj Nyman]. PMID- 9393133 TI - ["Take over home care". District nurses write a letter to the community]. PMID- 9393134 TI - [Employers refuse to explain wage differences]. PMID- 9393135 TI - [Can faith move mountains?]. PMID- 9393136 TI - [Tough fight for Swedish registration]. PMID- 9393137 TI - [Fishing for votes and pulling the wool over someone's eyes]. PMID- 9393138 TI - [Tests which can prevent thrombosis. "They are too expensive", say physicians]. PMID- 9393139 TI - [Nurses should stick out their chin themselves. Interview by Birgitta Dalenstam]. PMID- 9393140 TI - [In memory of a child. Interview by Ingela Bjorck]. PMID- 9393141 TI - [Life situation in children with cancer. Questionnaire helps in determining quality of life]. PMID- 9393142 TI - [Many seek new education in cytology]. PMID- 9393143 TI - ["How do I do that?" Action provides relatives quick contact with health care]. PMID- 9393144 TI - [Chief physician cautioned for lack of management responsibility]. PMID- 9393145 TI - [Take dizziness seriously. Interview by Bodil Sundvall]. PMID- 9393146 TI - ["We do not put up vendettas". Members do not always get legal assistance from the association]. PMID- 9393147 TI - [New vision will guide association. "We want to describe how it will be in 10 years. Interview by Kaj Nyman]. PMID- 9393148 TI - [Petra will look after association's students' issues. Interview by Jan Thomasson]. PMID- 9393149 TI - The importance of patient registration and processing. PMID- 9393150 TI - Radiation dosimetry in diagnostic radiology. PMID- 9393151 TI - A database program for the management of staff scheduling in a radiology department. AB - OBJECTIVE: Our objective is to report how an inexpensive computer database program (Filemaker Pro, version 3.0, for Macintosh) can be used to manage work schedules and optimize staff use in a radiology department. CONCLUSION: Using this report in conjunction with the manufacturer's documentation, one can adapt this database program to any scheduling situation. PMID- 9393152 TI - Contrast-enhanced CT of intrahepatic and hilar cholangiocarcinoma: delay time for optimal imaging. AB - OBJECTIVE: The purpose of this study was to determine the optimal time for obtaining delayed images with contrast-enhanced CT in patients who have intrahepatic or hilar cholangiocarcinoma. SUBJECTS AND METHODS: CT studies were performed in 25 consecutive patients with proven cholangiocarcinoma, including six patients who had undergone radiotherapy or chemotherapy. Dynamic images of the liver were obtained after 150 ml of IV contrast material was administered at 3 ml/sec. Delayed CT images were then obtained at 10, 20, and 30 min. Tumor-liver attenuation difference was determined quantitatively for each time period. Images were qualitatively evaluated by three observers for attenuation of the tumor (hypoattenuating, isoattenuating, or hyperattenuating) relative to the liver. Observer confidence for tumor detection was graded on a four-point scale. Dynamic and delayed images were compared for tumor conspicuity. RESULTS: On dynamic images, 18 tumors (72%) were hypoattenuating, six (24%) were isoattenuating, and one was heterogeneous. On delayed images, 15 (60%) of these 25 tumors were isoattenuating and nine (36%) were hyperattenuating compared with the liver. Tumor-liver attenuation difference was greatest on dynamic studies (p < .01) and did not differ significantly among the three delay times (p > .20). All tumors seen on delayed images were also seen on dynamic images; however, in three patients (12%), the confidence level for presence of tumor was better on delayed than on dynamic images. Confidence levels for presence of tumor did not vary significantly among the three delay times. Attenuation values on dynamic and delayed images did not differ for the groups of patients who had or had not undergone prior radiotherapy or chemotherapy (p > .05). CONCLUSION: In the evaluation of hilar or intrahepatic cholangiocarcinoma, delayed CT images are helpful for tumor characterization and may improve observer confidence for the presence of tumor. The optimal time for acquisition of delayed images is 10-20 min after contrast media injection. PMID- 9393153 TI - HASTE MR cholangiography in the evaluation of hilar cholangiocarcinoma. AB - OBJECTIVE: The objective of this study was to determine the usefulness of MR cholangiography using the half-Fourier acquisition single-shot turbo spin-echo sequence in the examination of patients with hilar cholangiocarcinoma. CONCLUSION: Half-Fourier acquisition single-shot turbo spin-echo MR cholangiography is a useful, noninvasive adjunct to other imaging techniques, particularly MR imaging, in the evaluation of hilar cholangiocarcinoma. MR cholangiography allows rapid visualization of the biliary tract without instrumentation and, therefore, without the risk of inducing sepsis in a patient with ductal obstruction. In the six patients presented. MR cholangiography allowed for determination of the proximal extent of disease and assessment of resectability and delineated the duct both proximal and distal to the stricture and isolated ductal obstructions. MR cholangiography provides three-dimensional images of the biliary tract that facilitate planning of surgery, palliative drainage, and radiation therapy. PMID- 9393154 TI - Detection of hepatocellular carcinoma in patients with cirrhosis: MR imaging versus angiographically assisted helical CT. AB - OBJECTIVE: The purpose of our study was to compare the combination of conventional spin-echo, phase-shift gradient-recalled echo (GRE), and triple phasic dynamic GRE MR imaging with the combination of helical CT hepatic arteriography (CTA) and CT performed during arterial portography (CTAP) in the preoperative detection of hepatocellular carcinoma (HCC). MATERIALS AND METHODS: Thirty-seven patients with cirrhosis underwent MR imaging and angiographically assisted CT imaging. Paired T1- and T2-weighted spin-echo images, paired in-phase and out-of-phase GRE images, triple-phasic dynamic GRE images, the combined MR images, and the paired CTA and CTAP images were retrospectively and independently reviewed by three radiologists. Image review was done on a segment-by-segment basis. Of the 280 liver segments, 58 segments contained 79 HCCs that were 0.5-8.0 cm (mean, 2.0 cm) in diameter. The diagnostic value of each pair of images was rated by means of receiver operating characteristic curve analysis. RESULTS: The diagnostic accuracy of combined CTA and CTAP (mean area under the receiver operating characteristic curve [Az] = 0.94) was significantly better than that of spin-echo (Az = 0.86, p < .0001), phase-shift GRE (Az = 0.83, p < .0001), dynamic GRE (Az = 0.85, p < .0001), and all combined (Az = 0.91, p < .001) MR imaging. The relative sensitivity of combined CTA and CTAP (89%) was also significantly (p < .0005) better than that of the combined MR imaging (75%). CONCLUSION: Angiographically assisted helical CT imaging was superior to MR imaging combined with conventional spin-echo, phase-shift GRE, and triple-phasic dynamic GRE techniques in the detection of HCC in patients with cirrhosis. The noninvasive dedicated combined MR imaging could not obviate invasive angiographically assisted CT imaging. Combined CTA and CTAP is recommended, especially in the preoperative examination of patients with HCC. PMID- 9393155 TI - Placement of metallic stents for treatment of postoperative biliary strictures: long-term outcome in 25 patients. AB - OBJECTIVE: This study was undertaken to evaluate the results of our 7-year experience with Gianturco-Rosch metallic stents, used for the management of postoperative biliary strictures. SUBJECTS AND METHODS: From January 1989 to April 1995, self-expanding Gianturco-Rosch metallic stents were placed in 25 patients with postoperative bile duct stenosis. All patients had a history of bile duct injury during cholecystectomy. Twenty-four patients had a conventional open cholecystectomy and one patient had a laparoscopic cholecystectomy. Eight patients had stenosis at the level of the common bile duct. The other 17 patients, who had undergone surgical repair of the bile duct, had a stricture at the level of the hepaticojejunostomy. These anastomotic strictures recurred after simple cholangioplasty. Patients were monitored for 9-84 months (mean, 55 months). Treatment was considered successful if the initial stenosis did not recur. Treatment was considered a failure if the initial stenosis recurred within the stent. RESULTS: Two patients had early complications: one had bile pleural effusion, treated with percutaneous drainage, and the other had arterial hemobilia, treated with embolization. Eighteen (72%) of 25 patients had no recurrence of the initial strictures. Among these patients, 11 had no further symptoms of biliary obstruction and seven, all with strictured hepaticojejunostomies, had recurrent episodes of cholangitis caused by secondary sclerosing cholangitis or intrahepatic stone formation. Seven (28%) of 25 patients had recurrence of the initial stenoses, causing repeated episodes of cholangitis. Among these seven patients, six had common bile duct stenoses and one had an anastomotic stricture. Recurrent biliary obstruction was treated surgically or with percutaneous methods, despite the presence of the metallic stent. CONCLUSION: Gianturco-Rosch stent placement should be considered in patients with postoperative bile duct stenoses in whom another operation is not indicated and cholangioplasty has failed. The results are better in patients who have hepaticojejunostomy strictures rather than common bile duct strictures. Overall, a long-term recurrence rate of cholangitis of more than 50% of patients was seen because of recurrence of the original stenosis or intrahepatic bile duct obstruction. PMID- 9393156 TI - Benign biliary strictures associated with recurrent pyogenic cholangitis: treatment with expandable metallic stents. AB - OBJECTIVE: The purpose of this study was to determine the long-term effectiveness of expandable metallic stents in benign biliary strictures associated with recurrent pyogenic cholangitis and the differences in primary patency of the various types of stents deployed. SUBJECTS AND METHODS: During a 20-month period, 26 metallic stents (19 Gianturco-Rosch Z stents and seven Strecker stents) were used to treat benign biliary strictures associated with recurrent pyogenic cholangitis in 23 patients (11 men and 12 women; mean age, 42 years; range, 30-78 years). Insertion routes were percutaneous transhepatic biliary drainage tracts for 16 stents, T-tube tracts for seven stents, and retrograde endoscopic routes for three stents. The deployed locations were common hepatic or common bile ducts for 11 stents, right or left hepatic ducts for 10 stents, and segmental ducts for five stents. RESULTS: The initial technical success rate was 100%. Two stents in one patient migrated spontaneously. Primary stent patency for the remaining 24 stents was 34 months (range, 3-58 months). Primary stent patency of the Gianturco Rosch Z and Strecker stents was 50 and 10 months, respectively (p < .05). Primary stent patency for the intrahepatic and extrahepatic ducts was 50 and 18 months, respectively (p = .05). Primary patency rates for all stents at 6, 12, 24, and 36 months were 92%, 75%, 67%, and 46%, respectively. The causes of stent obstruction were recurrent stone or sludge in eight stents and epithelial hyperplasia in five stents. CONCLUSION: We believe that metallic stent placement is not an effective long-term treatment technique for benign biliary stricture associated with recurrent pyogenic cholangitis. PMID- 9393157 TI - Dissecting room. PMID- 9393158 TI - Displaced metallic biliary stents: technique and rationale for interventional radiologic retrieval. AB - OBJECTIVE: We investigated the spontaneous course and the possibility of transhepatic removal of displaced biliary stents. MATERIALS AND METHODS: Displaced biliary stents were observed in 11 patients (13-75 years old) between October 1988 and August 1996. Stent types included the Palmaz stent (n = 3), Wallstent (n = 3), and the Strecker stent (n = 5). Reasons for stent displacement included primary misplacement (n = 4), dislocation due to transhepatic endoscopy with biopsy (n = 2), dislocation resulting from a recanalization maneuver in stent occlusion (n = 3), and unknown causes (n = 2). In three cases, the stent was displaced into the proximal bile duct system. Seven patients had primary malignancy. RESULTS: Eight of 11 displaced biliary stents were removed transhepatically. Extraction was performed using either a wire loop (n = 4) or forceps (n = 4). No complications occurred. In the remaining three patients, whose stents were displaced into the intestine, no invasive action was taken. In one of these patients, a Palmaz stent was passed spontaneously after 1 week. In the second of these patients, a 6 cm Wallstent remained innocuously at a position in the right lower abdomen, and the patient died as a result of malignancy. In the third patient, who had a 10-cm Wallstent, an abscess developed in the stent region 4 months after displacement and resulted in formation of an ileocutaneous fistula. CONCLUSION: Transhepatic extraction of displaced biliary stents is technically possible, even in the case of rigid stents such as the Palmaz stent. Because of the risk of intestinal perforation, displaced stents should be removed. PMID- 9393159 TI - Outcome of 17 patients with portal venous gas detected by CT. AB - OBJECTIVE: Portal venous gas has historically been associated with a poor prognosis. The clinical outcomes of 17 patients with portal venous gas detected by CT were reviewed. CONCLUSION: Portal venous gas seen on CT was associated with a 71% survival rate. Patients with a history of vascular surgery or bowel obstruction may have a higher mortality, and careful clinical evaluation is mandatory in this patient population. PMID- 9393160 TI - Solitary hepatic lesions with a hypoechoic rim: value of color Doppler sonography. AB - OBJECTIVE: A hypoechoic rim around a focal liver lesion as revealed by conventional sonography may be present in malignant liver lesions as well as in benign liver lesions. This study evaluated the potential of color Doppler sonography in differentiating various focal liver lesions with a hypoechoic rim. CONCLUSION: Color Doppler sonography may be helpful in distinguishing focal nodular hyperplasia (FNH) from other focal liver lesions. The characteristic finding of blood flow within the hypoechoic rim of FNH is most likely caused by small vessel abnormalities that have previously been described for FNH. PMID- 9393161 TI - Successful transjugular intrahepatic portosystemic shunt creation in a patient with polycystic liver disease. PMID- 9393162 TI - Multiplanar helical CT enterography in patients with Crohn's disease. AB - OBJECTIVE: The purpose of this study was to assess the feasibility and usefulness of helical CT with multiplanar reformations in revealing complications in patients with Crohn's disease. SUBJECTS AND METHODS: Twenty-two patients with Crohn's disease and clinically suspected complications underwent helical CT enterography. The imaging protocol began with the administration of a large volume (1600 ml) of oral contrast material followed by helical scanning with axial and multiplanar two-dimensional coronal projections. Three independent observers assessed the adequacy of bowel opacification and the contribution of two-dimensional coronal projections to the interpretation of axial images. CT enterography was compared with conventional barium studies in 14 patients. Statistical analysis included repeated measures analysis of variance, the Wilcoxon signed-rank test, and the McNemar test. RESULTS: The large oral contrast medium dose was well tolerated and provided optimal bowel opacification in 21 of 22 patients. The addition of multiplanar to conventional axial images did not reveal additional abnormalities; however, multiplanar imaging significantly improved observers' confidence in their interpretation of imaging and in their assessment of the extent of bowel wall thickening (p < .01). Interobserver agreement was 78%. Findings on helical CT were comparable with those on barium studies in nine of 14 patients, superior to those on barium studies in four patients, and inferior in one patient. CONCLUSION: CT enterography is a useful technique for bowel imaging. In patients with complicated Crohn's disease, multiplanar imaging improves confidence in assessing the presence and extent of disease. CT enterography is complementary and often superior to conventional barium studies. PMID- 9393163 TI - Percutaneous radiologic and endoscopic gastrostomy: a 3-year institutional analysis of procedure performance. AB - OBJECTIVE: The study was designed to evaluate the safety, efficacy, and usefulness of the performance of percutaneous radiologic (PRG) and endoscopic (PEG) gastrostomy. MATERIALS AND METHODS: This study involved a retrospective review of 182 percutaneous gastrostomy procedures (68 PRG, 114 PEG) performed over a 3-year period. Parameters analyzed included technical success, procedure duration, anesthetic requirements, incidental findings on endoscopy, and complications. RESULTS: The success rate for tube placement was higher for PRG than for PEG (100% versus 95%). PRG was subsequently performed in four of six patients in whom PEG procedures failed. Mean procedure duration was shorter for PRG than for PEG (32.9 min versus 39.1 min, p < .05). PRG was performed without conscious sedation (i.e., local anesthesia only) more frequently than was PEG (25% versus 0%, p < .001). The mean volume of doses of midazolam hydrochloride administered during PRG was two thirds of the volume of doses used during PEG. Incidental abnormalities were detected in 32 (30%) of the successful PEG procedures, 11 (10%) of which resulted in biopsy or medical therapy. No biopsy specimen showed evidence of malignancy. One (0.9%) patient received treatment other than gastric acid medication. Three (3%) major postprocedural complications occurred immediately after PEG and none (0%) occurred after PRG. CONCLUSION: The significant advantages of PRG over PEG included higher success rates, shorter procedure duration, and less conscious sedation required. PRG was also successful with patients for whom PEG failed. PMID- 9393164 TI - The role of parity and hysterectomy on the development of pelvic floor abnormalities revealed by defecography. AB - OBJECTIVE: The purpose of our study was to assess the role of multiparity and pelvic surgery, especially hysterectomy, on pelvic floor dysfunction as diagnosed on defecography. MATERIALS AND METHODS: Three hundred fifty-four women who underwent defecography between 1986 and 1996 were asked to provide information regarding obstetric history and pelvic surgery. Responses were obtained from 272 women (response rate, 77%). Their presenting symptoms ranged from incontinence to constipation and obstructed defecation. Historical data were correlated with incidence of defecographic abnormalities that included rectocele, enterocele, rectal prolapse, incontinence, excessive pelvic floor descent, and dyskinetic puborectal muscle. RESULTS: Women with three or more birth deliveries were more likely to have incontinence (48% versus 36%, p = .05) and excessive pelvic floor descent (26% versus 17%, p = .07) than women who had delivered fewer children. Women undergoing hysterectomy before defecography were more likely to have enterocele (40% versus 25%, p = .009) and excessive pelvic floor descent (25% versus 15%, p = .04) than women who had never undergone hysterectomy. CONCLUSION: Our findings confirm the common belief that trauma from childbirth or hysterectomy contributes to the development of defecation disorders. PMID- 9393166 TI - Double-contrast examination of the colon without decubitus films. PMID- 9393165 TI - Sonography of intraabdominal gas collections. PMID- 9393167 TI - Characteristic angiographic appearance of inverted Meckel's diverticulum. PMID- 9393169 TI - The use of unenhanced helical CT to evaluate suspected renal colic. PMID- 9393168 TI - Dual-phase helical CT of the kidney: value of the corticomedullary and nephrographic phase for evaluation of renal lesions and preoperative staging of renal cell carcinoma. AB - OBJECTIVE: Our objective was to evaluate early-phase unenhanced and late-phase contrast-enhanced helical CT in revealing renal lesions and staging renal cell carcinomas. SUBJECTS AND METHODS: Contrast-enhanced helical CT of the kidneys was performed in 145 patients who also underwent unenhanced CT. Contrast-enhanced CT was performed in the corticomedullary phase (CMP) and nephrographic phase (NP). A total of 173 lesions in 96 patients were proven histologically or cytologically. The performance of helical CT in the three study groups was compared: unenhanced and CMP enhancement, group 1; unenhanced and NP enhancement, group 2; unenhanced, CMP enhancement, and NP enhancement, group 3. Among the parameters evaluated were the sensitivity for helical CT of all 173 renal lesions and the sensitivity and specificity for the 90 malignant tumors. Also, the preoperative CT staging of the 76 renal cell carcinomas was correlated with the pathologic specimens. RESULTS: The sensitivity for detection of all renal lesions in group 1 (84%) was significantly less than in groups 2 and 3 (97% and 100%). The specificity and accuracy of helical CT in revealing renal cell carcinomas were significantly higher (p < .05) in group 3 (95% and 95%, respectively) than in groups 1 (93% and 92%, respectively) and 2 (89% and 91%, respectively). Two renal cell carcinomas were overlooked by the interpreters of the helical scans in group 1. The accuracy of preoperative CT staging of renal cell carcinomas was significantly better (p < .05) in group 3 (91%) than in groups 1 (82%) and 2 (86%). CONCLUSION: When patients underwent unenhanced helical CT, CMP helical CT, and NP helical CT, we achieved a better rate of detection and characterization of renal lesions and better preoperative staging of renal cell carcinomas than when we used either CMP helical CT or NP helical CT alone. PMID- 9393170 TI - Unilateral tubular obstruction: an unusual complication of urographic contrast media. AB - OBJECTIVE: Our purpose was to report our experience with four patients with renal colic who had a rare unilateral complication from urographic contrast media, with a dense and striated nephrogram affecting only the healthy kidney. All cases were reversible. CONCLUSION: An unusual complication of urography with a characteristic radiologic appearance is presented. This complication was transient and reversible in our patients. Unilateral tubular blockage induced by a contrast agent must be considered when the contralateral kidney shows evidence of ureteral obstruction. PMID- 9393171 TI - Radiologic insertion of subcutaneous nephrovesical stent for inoperable ureteral obstruction. PMID- 9393172 TI - Comparison of real-time virtual and fiberoptic bronchoscopy in patients with bronchial carcinoma: opportunities and limitations. AB - OBJECTIVE: Both helical CT and fiberoptic bronchoscopy are used in the staging of pulmonary tumors for therapeutic decision making. The improved resolution offered by helical CT has led to the clinical use of three-dimensional reconstruction techniques such as virtual bronchoscopy. We tested this new simulated endoscopic view of inner organ surfaces and compared it with corresponding fiberoptic examinations of the tracheobronchial system. SUBJECTS AND METHODS: Twenty patients with malignancies of the lung and mediastinum were examined with both virtual bronchoscopy and fiberoptic bronchoscopy. Both examinations were reviewed by radiologists and surgeons familiar with fiberoptic bronchoscopy. Virtual bronchoscopy was calculated and reconstructed from the cross-sectional images on a separate workstation. Stenoses and tumor infiltration were classified from the fiberoptic examination. These results were compared with the virtual bronchoscopy findings. RESULTS: Virtual bronchoscopy of diagnostic quality was achieved in 19 of 20 patients. High-grade stenoses were revealed equally well with virtual and fiberoptic techniques. Virtual bronchoscopy offered the advantage of being able to visualize areas beyond even high-grade stenoses. However, on virtual bronchoscopy discrete infiltration or extraluminal impression was not visible in five patients. In another patient, strong heart pulsation produced motion artifacts that prevented evaluation of the reconstruction. CONCLUSION: Virtual bronchoscopy represents a new noninvasive method for evaluating helical CT findings. In comparison with fiberoptic bronchoscopy, virtual bronchoscopy offers the advantage of being able to visualize areas beyond even high-grade stenoses. In addition to the limited view of fiberoptic bronchoscopy, extraluminal causes of lumen compressions can be analyzed in the cross-sectional images and evaluated together with the virtual representation. However, it was not possible to detect small infiltrations with virtual bronchoscopy. This new representation of helical CT data might be helpful for postoperative follow-up examinations, such as after stent implantation, and can be carried out without additional risk to the patient. Radiologists do need special fiberoptic bronchoscopy knowledge and experience with three-dimensional-reconstructions to differentiate between real stenoses and artificial stenoses that might be caused by pulsation artifacts. PMID- 9393173 TI - Altered intravascular contrast material flow dynamics: clues for refining thoracic CT diagnosis. AB - The ability of helical thoracic CT to acquire data rapidly has enabled radiologists to use the dynamics of contrast medium flow more effectively to diagnose subtle abnormalities. In general, these abnormal flow dynamics manifest in two ways: diverted hyperdense venous opacification and an altered temporal relationship of vascular opacification. When observed, these findings serve as clues for the detection of elusive anatomic and physiologic abnormalities. PMID- 9393174 TI - Sonographic guidance of mediastinal biopsy: an effective alternative to CT guidance. AB - OBJECTIVE: Our objective was to determine the diagnostic and cost efficacy of sonographically guided mediastinal biopsy as an alternative to CT-guided mediastinal biopsy and to review our biopsy triage experience in switching from CT to sonography. CONCLUSION: Sonography is as safe and accurate as CT and is 25% less costly than CT. Sonography proved particularly valuable for identifying vessels and perfused tissue and for permitting upright biopsy positions in dyspneic patients. When using our triage criteria, radiologists should find sonographically guided mediastinal biopsy to be an attractive alternative to CT guided mediastinal biopsy in most patients. PMID- 9393175 TI - Detection of pulmonary nodules with helical CT: comparison of cine and film-based viewing. AB - OBJECTIVE: The purpose of our study was to determine whether cine viewing of helical CT scans of the chest improves the detection of pulmonary nodules in patients with known extrathoracic malignancy. SUBJECTS AND METHODS: Identical helical CT studies of the chest of 60 patients with known extrathoracic malignancy were reviewed for detection of pulmonary nodules. Four radiologists interpreted the helical CT studies. Pulmonary nodules were divided into four groups according to maximum diameter: group 1, nodules smaller than or equal to 5 mm; group 2, nodules larger than 5 mm but smaller than or equal to 10 mm; group 3, nodules larger than 10 mm but smaller than or equal to 20 mm; group 4, nodules larger than 20 mm. Interpreters also assigned a lesion conspicuity score of pulmonary nodules based on a four-point scale: one point for poor visibility, two points for adequate visibility, three points for good visibility, and four points for excellent visibility. Static film-based images printed on a laser printer were viewed on a light box. Cine viewing of helical CT scans from the same examinations was done on a commercially available workstation. The number, diameter, and conspicuity scores of pulmonary nodules detected at lung window settings were documented. RESULTS: Interpreters saw 266 nodules on cine viewing, whereas 237 nodules were seen with static film-based viewing. A significantly higher percentage of nodules that were smaller than or equal to 5 mm in diameter was found with cine viewing (n = 106) than with static film-based viewing (n = 81) (p < .05). Cine viewing (n = 105) also allowed a slightly but not significantly higher detection rate of nodules that were larger than 5 mm but smaller than or equal to 10 mm in diameter than did static film-based viewing (n = 101). We found no differences between cine (n = 55) and static film-based viewing (n = 55) in the detection of pulmonary nodules that were larger than 10 mm in diameter. The mean conspicuity score of nodules was significantly higher with cine viewing (2.9 +/- 0.2) than with film-based viewing (2.4 +/- 0.2) (p < .05). CONCLUSION: Cine viewing of helical CT scans significantly increases the detection rate of pulmonary nodules that are smaller than or equal to 5 mm in diameter. However, we found no significant difference between cine and film-based viewing in the detection rate of pulmonary nodules that were larger than 5 mm in diameter. The advantages of cine viewing may be attributed to both the larger image size and the ability to scroll through images for improved differentiation between vessels and nodules. PMID- 9393176 TI - Interventional drainage of appendiceal abscesses in children. AB - OBJECTIVE: This study was undertaken to validate the outcome of interventional drainage and IV antibiotics for the treatment of appendiceal abscesses in children. MATERIALS AND METHODS: Between March 1991 and March 1995, 46 children with one or more intraabdominal appendiceal abscesses were seen at a tertiary care referral center. The children received IV antibiotics and underwent interventional drainage of their collections. All procedures were performed in the radiology intervention suite using IV sedation and, in one patient, a general anesthetic. All patients were followed up for at least 1 year. RESULTS: The 46 patients underwent 64 procedures. These included the insertion of 34 percutaneous drainage catheters, the insertion of 25 transrectal drainage catheters, and five needle aspirations. Four patients did not respond to their initial treatment and required surgery. One patient developed a colonic fistula that resolved spontaneously. CONCLUSION: Successful treatment of 42 patients (91%) justifies image-guided drainage and IV antibiotics as appropriate management for appendiceal abscesses in children. PMID- 9393177 TI - Fat necrosis after trauma: a benign cause of palpable lumps in children. AB - OBJECTIVE: Our objective is to describe the clinical entity and MR imaging appearance of fat necrosis after trauma, a benign cause of palpable soft-tissue lesions in children. A related objective is to establish MR imaging criteria that can be used reliably to differentiate this entity from other more serious causes of soft-tissue masses. CONCLUSION: Fat necrosis after trauma is a common cause of palpable lumps in children and has a benign course on long-term follow-up. When characteristic MR imaging findings are seen, conservative therapy is appropriate. PMID- 9393179 TI - Endodermal sinus tumor of the vagina. PMID- 9393178 TI - Spectrum of imaging findings in pediatric hydatid disease. PMID- 9393181 TI - Sonography of the breast: controversies and opinions. PMID- 9393180 TI - Comparison of duodenal intubation techniques during conversion of gastrostomy to gastrojejunostomy tubes in children. PMID- 9393182 TI - Metallic particles on mammography after wire localization. PMID- 9393183 TI - Granulocytic sarcoma (chloroma) of the breast: sonographic findings. PMID- 9393184 TI - Posterolateral stabilizers of the knee: anatomy and injuries assessed with MR imaging. PMID- 9393185 TI - Carpal tunnel syndrome: are the MR findings a result of population selection bias? AB - OBJECTIVE: Previous descriptions of MR imaging of carpal tunnel syndrome used limited study populations and volunteers as controls. We reevaluated these descriptions to determine their sensitivity and specificity when applied to a large consecutive clinical series in which the incidence of carpal tunnel syndrome was small. SUBJECTS AND METHODS: In 196 consecutive wrists for which supplemental axial conventional spin-echo T1-weighted and fast spin-echo T2 weighted images were obtained at 1.5 T with a dedicated wrist coil, 165 studies were available for review. Previously described signs of carpal tunnel syndrome such as proximally increased size, flattening of the median nerve, increased median nerve signal intensity, flexor tenosynovitis, retinacular bowing, decreased deep tendon fat, and deep palmar bursitis were retrospectively and independently evaluated by two observers who were unaware of patient diagnosis. RESULTS: None of the previously described signs was sensitive for the diagnosis of carpal tunnel syndrome. However, specificity was high for retinacular bowing (94%), median nerve flattening (97%), and deep palmar bursitis (95%). CONCLUSION: Most previously described MR imaging signs of carpal tunnel syndrome are insensitive and nonspecific. Exceptions include retinacular bowing, median nerve flattening, and deep palmar bursitis, which in our study proved to have specificities greater than or equal to 94%. PMID- 9393186 TI - A comparison of whole-body turboSTIR MR imaging and planar 99mTc-methylene diphosphonate scintigraphy in the examination of patients with suspected skeletal metastases. AB - OBJECTIVE: This study was undertaken to compare whole-body turbo short inversion time inversion recovery MR imaging and 99mTc-methylene diphosphonate planar scintigraphy in the examination of patients with suspected skeletal metastases. SUBJECTS AND METHODS: Twenty-five patients with known or suspected skeletal metastatic disease underwent both whole-body turbo short inversion time inversion recovery MR imaging and whole-body 99mTc-methylene diphosphonate scintigraphy. RESULTS: MR imaging revealed metastases at 57 of 175 possible sites (sensitivity, 96.5%, specificity, 100%; positive predictive value, 100%). Scintigraphy revealed metastases at 43 of 175 possible sites (sensitivity, 72%; specificity, 98%; positive predictive value, 95%) (McNemar test, 0.01; p = .016). Discrepancies in skeletal evaluation by whole-body MR imaging and scintigraphy were observed in six (24%) of 25 patients. Soft-tissue abnormalities were identified in 13 (52%) of 25 patients with MR imaging alone. CONCLUSION: Preliminary results suggest that whole-body MR imaging is an effective method of examining patients with suspected skeletal metastases, with better sensitivity than conventional planar 99mTc-methylene diphosphonate scintigraphy. PMID- 9393187 TI - FDG PET in head and neck cancer. AB - In our extensive experience with FDG PET imaging in head and neck cancer, we have found the technique to be of high accuracy but of limited usefulness. This seeming paradox arises from several causes. Competing techniques such as CT, MR imaging, and even clinical examination already have good accuracy. In addition, high-resolution studies such as CT and MR imaging provide information required for treatment planning that is unavailable from FDG PET images. The high cost of FDG PET militates against its use in this setting, in which only a small marginal gain can be expected. In the special problem areas in which FDG PET might be expected to offer unique advantages, such as screening for second primary lesions, searching for unknown primary lesions, or differentiating benign salivary rumors from malignant lesions, the results of FDG PET have been disappointedly poor. Of these special problem areas, only the question of accuracy in finding occult primary lesions appears unresolved and in need of further study. The single application in which FDG PET appears to be advantageous is the posttherapy setting. In this setting, the technique is definitely superior to alternative methods of determining tumor recurrence and differentiating posttherapy sequelae such as radiation necrosis from tumor recurrence. We believe that considerable opportunity remains for further research on the use of FDG PET in head and neck cancer. Other agents such as 11C-methionine for example, might improve the diagnostic accuracy of FDG PET in some of the problem areas that we have identified, such as the early postirradiation period. We currently have such a study under way. Also, because FDG PET offers a unique way to measure tumor metabolism, further investigation of the use of FDG PET tracers to evaluate various biologic parameters such as proliferation rates or tumor hypoxia are needed. Such studies could provide a noninvasive technique to identify which fractionation schemes or combinations of therapy might be useful for individual patients. A final caveat is in order. Although our findings of the usefulness (and lack thereof) of FDG PET in head and neck cancer may be disappointing to many, these results should not be generalized to other applications of FDG PET in oncology. Each tumor type and setting presents its own specific problems, and in some instances FDG PET offers unique advantages over other imaging techniques. A good example is the setting of primary lung cancer, in which FDG PET appears clearly superior to all other methods of pretherapy screening [19-20]. PMID- 9393188 TI - Comparison of double-phase 99mTc-sestamibi with 123I-99mTc-sestamibi subtraction SPECT in hyperparathyroidism. AB - OBJECTIVE: Our purpose was to compare double-phase 99mTc-sestamibi single-photon emission computed tomography (SPECT) and simultaneous 123I-99mTc-sestamibi subtraction SPECT for preoperative localization of hyperfunctioning parathyroid tissue in patients with primary hyperparathyroidism. SUBJECTS AND METHODS: Fifteen patients with primary hyperparathyroidism underwent preoperative double phase 99mTc-sestamibi SPECT and simultaneous 123I-99mTc-sestamibi subtraction SPECT imaging. At surgery, the location, weight, and histopathologic evaluation of all identified parathyroid glands were recorded. RESULTS: At surgery, 17 parathyroid adenomas and 37 normal parathyroid glands were identified. The sensitivity, specificity, and diagnostic accuracy for the detection of parathyroid adenomas were 88%, 97%, and 94%, respectively, for simultaneous 123I 99mTc-sestamibi subtraction SPECT and 53%, 86%, and 76%, respectively, for double phase 99mTc-sestamibi SPECT. The differences in sensitivity and diagnostic accuracy were statistically significant (p = .031 and p = .016, respectively). CONCLUSION: Compared with double-phase 99mTc-sestamibi SPECT, simultaneous 123I 99mTc-sestamibi subtraction SPECT is a superior imaging study for the preoperative localization of hyperfunctioning parathyroid tissue. PMID- 9393189 TI - Whole-body positron emission tomography: normal variations, pitfalls, and technical considerations. PMID- 9393190 TI - Can radiologists accurately predict preepiglottic space invasion with MR imaging? AB - OBJECTIVE: The purpose of this study was to evaluate whether observers of MR imaging can accurately predict invasion of the preepiglottic fat (PEF) in patients with oropharyngeal and supraglottic laryngeal squamous cell carcinoma. MATERIALS AND METHODS: For 41 patients with pathologically proven squamous cell carcinoma of the oropharynx and supraglottic larynx, we retrospectively analyzed their MR images for the presence or absence of PEF neoplastic invasion. Unenhanced T1-weighted, fat-suppressed T2-weighted, and contrast-enhanced fat suppressed T1-weighted scans were analyzed independently by two neuroradiologists who were unaware of the surgical findings. Proof of diagnosis was determined by pathologic analysis, intraoperative assessment, or both. RESULTS: Sixteen patients had neoplastic infiltration of the PEF. All infiltration was correctly predicted by the two observers of MR imaging, resulting in a sensitivity of 100%. Twenty-five patients had no invasion of the PEF by pathologic or surgical evaluation or both. Of these patients, negative findings were correctly predicted on MR imaging in 21 patients, whereas positive findings were incorrectly predicted on MR imaging in the remaining four patients, resulting in a specificity of 84% and an accuracy of 90%. In two of the four false-positive cases, effacement of the fat in the preepiglottic space by large tumors was mistaken for invasion. In a third patient, spread to the paraglottic space was mistaken for PEF extension. In the fourth false-positive case, glandular tissue along the ventral epiglottis may have been mistaken for tumor. The observers believed that unenhanced sagittal and axial T1-weighted scans were particularly useful because fat saturation artifacts may degrade T2-weighted and contrast enhanced T1-weighted scans. CONCLUSION: Unenhanced T1-weighted MR images are highly sensitive for neoplastic infiltration of the preepiglottic space in patients with oropharyngeal and supraglottic laryngeal carcinoma who are at risk for such spread. Identification of PEF invasion is important because it affects prognosis and may affect surgical management. PMID- 9393191 TI - Postoperative radiographic assessment of the Combi 40 cochlear implant. AB - OBJECTIVE: The aims of this study were to establish a plain radiographic technique for the assessment of the postoperative appearance, position, and insertion depth of the Combi 40 cochlear implant and to correlate the radiologic findings with surgical reports. SUBJECTS AND METHODS: In an experimental study, an electrode of the Combi 40 device was inserted into the cochlea of a cadaveric skull. Digital radiographs were obtained in a modified Chausse III projection, in which the skull was placed supine on the radiography table with the infraorbitomeatal line strictly perpendicular to the film cassette. The skull was then rotated 30 degrees away from the side to be examined, and the central X-ray beam was angled 15 degrees cephalad to the infraorbitomeatal line. On these radiographs, the point of cochleostomy was marked by a needle tip and was projected inferior to the vestibule and on a line drawn through the superior semicircular canal and the vestibule. The appearance and position of the electrode was evaluated. An electrode was defined as completely inserted if all electrode contacts projected medial to the line drawn through the superior semicircular canal and the vestibule. We also studied cochlear implant insertion of the Combi 40 device in 37 patients. Postoperative digital radiographs of these patients were obtained and analyzed for the criteria as defined in the cadaveric study. In addition, the insertion depth of the electrode and the angle of insertion were measured on the radiographs. This depth was correlated with depth of insertion as estimated at surgery. RESULTS: The cadaveric study showed that the completely inserted electrode was seen on radiographs as a nonoverlapping spiral within the cochlea. All electrode contacts projected medial to the line drawn through the superior semicircular canal and the vestibule. In all 37 patients, the electrode could be seen without overlapping. According to our criteria, a completely inserted electrode was seen in 32 patients. In these patients, the insertion depth ranged from 21 to 34 mm and the angle of insertion ranged from 350 degrees to 810 degrees. In two patients, we saw a completely inserted electrode with a bend. In three patients, an incompletely inserted electrode was seen. Excellent correlation existed between the radiologic and surgical results with regard to insertion depth (r = .92). CONCLUSION: Digital radiographs obtained in the modified Chausse III projection allow clear depiction of the electrode and avoid overlapping. Such radiographs enable a reliable and accurate assessment of the position and insertion depth of the electrode of this new cochlear implant. Such images can serve as a baseline for further radiographic examinations when extrusion or slippage of the electrode is clinically suspected. PMID- 9393192 TI - Sonography of peripheral nerve tumors of the neck. AB - OBJECTIVE: Peripheral nerve tumors (PNTs) of the neck are uncommon and are frequently mistaken for lymph nodes. The purpose of this study is to describe the sonographic appearance of PNTs in the neck and determine features useful in differentiating PNTs from lymph nodes. CONCLUSION: PNTs of the neck have features that should alert the sonographer to the correct diagnosis. Features of value in differentiating PNTs from lymph nodes include a solitary, oval, hypoechoic mass with posterior enhancement and absence of an echogenic hilum. These features should lead to a careful search for the associated nerve to confirm the diagnosis. In addition, schwannomas have a vascular pattern indicating marked hypervascularity that can be obliterated with light pressure from the probe. PMID- 9393193 TI - Cerebral venography: comparison of CT and MR projection venography. AB - OBJECTIVE: The purpose of the study was to show equivalence or superiority of CT venography compared with the existing test of choice--MR venography--in the evaluation of dural sinus thrombosis and in the identification of cerebral veins and dural sinuses. MATERIALS AND METHODS: Twenty-four patients underwent both CT and MR venography of the intracranial venous circulation. Seventeen patients were examined for suspected dural sinus thrombosis. Four patients underwent projection venography to assess tumor invasion of a major dural sinus. The remaining three patients were examined for cavernous sinus thrombosis, arteriovenous malformation, and an elevated jugular bulb. Without knowledge of the patients' case histories, two radiologists evaluated each CT venogram and MR venogram. The radiologists then arrived at a consensus regarding the absence or presence of dural sinus thrombosis. Later, the radiologists conducted a second interpretation with knowledge of the patients' clinical histories during which time MR and CT venograms were compared with regard to the advantages and disadvantages of each imaging technique. In addition, the venograms were assessed for the presence of 12 different venous structure. Projection venograms were displayed using a maximum-intensity-projection (MIP) algorithm, and the individual source images were also evaluated. The CT venograms were also displayed using shell-MIP and integral display algorithms. RESULTS: Using MR venography, the two radiologists diagnosed dural sinus thrombosis in eight of the 17 patients with suspected dural sinus thrombosis. In these eight patients, the diagnosis was also made with CT venography. The diagnosis was confirmed by follow-up CT in four patients and by follow-up MR imaging in two patients. The MIP algorithm did not allow direct visualization of thrombus by either the CT or the MR imaging technique; however, the CT integral display algorithm enabled direct visualization of thrombus on the three-dimensional projection venograms. The systematic comparison of imaging techniques showed that CT venography reliably reveals all cerebral veins and sinuses when they are seen with MR venography. In addition, CT venography more frequently visualizes sinuses or smaller cerebral veins with low flow as compared with MR venography. CONCLUSION: Cerebral CT venography is superior to MR venography in the identification of cerebral veins and dural sinuses and is at least equivalent in the diagnosis of dural sinus thrombosis. CT venography is a viable alternative to MR venography in the examination of patients with suspected dural sinus thrombosis. PMID- 9393194 TI - Optic neuritis: imaging with magnetization transfer. AB - OBJECTIVE: Our objective was to prospectively examine the optic nerves in patients with clinically severe unilateral optic neuritis, using routine spin echo and magnetization transfer MR imaging. SUBJECTS AND METHODS: For 39 patients with such lesions, we calculated the magnetization transfer ratio along the involved intraorbital optic nerve and along the asymptomatic contralateral optic nerve in a mirror-image location. Magnetization transfer ratios were correlated with the imaging findings on routine spin-echo MR imaging. RESULTS: Magnetization transfer ratios were decreased in 33 of the 39 clinically symptomatic optic nerves, including 12 of the 18 clinically symptomatic optic nerves in which no abnormality was seen on routine spin-echo MR images obtained before and after administration of gadopentetate dimeglumine. CONCLUSION: Magnetization transfer imaging reveals intraorbital optic nerve abnormalities in patients with optic neuritis even when such lesions are otherwise occult on routine magnetization transfer imaging. PMID- 9393195 TI - MR imaging of intramedullary and intradural-extramedullary spinal cysticercosis. AB - OBJECTIVE: The purpose of our study was to retrospectively review the MR imaging findings in a group of patients with clinically proven cysticercosis involving the spinal cord, the spinal subarachnoid space, or both. MATERIALS AND METHODS: We retrospectively reviewed images of 16 patients with clinically diagnosed spinal cysticercosis to summarize the imaging characteristics. All patients underwent T1- and T2-weighted sagittal, axial, or both sagittal and axial MR imaging before i.v. administration of paramagnetic contrast media. Thirteen patients also underwent sagittal, axial, or both sagittal and axial T1-weighted MR imaging after i.v. gadolinium administration. In addition, all patients underwent cranial CT, MR imaging, or both to reveal possible evidence of cranial cysticercosis. RESULTS: MR imaging revealed isolated intradural-extramedullary involvement (n = 9), isolated intramedullary involvement (n = 3), combined intradural-extramedullary and intramedullary involvement (n = 3), and/or syringomyelia caused by infection and associated with chronic spinal arachnoiditis (n = 2). Evidence of intradural-extramedullary disease included cystic structures within the subarachnoid space or homogeneous sheetlike enhancement within the subarachnoid space over the surface of the spinal cord. Evidence of intramedullary disease included focal cystic lesions or syringomyelic cavitation of the spinal cord. All patients had evidence of simultaneous intracranial cysticercosis as shown on cranial CT, MR imaging, or both. CONCLUSION: In the absence of scolex visualization, cysticercotic involvement of the spinal cord or spinal subarachnoid space has a nonspecific appearance on MR imaging. On the basis of the findings in this group of patients, we believe that spinal cysticercosis is most often accompanied by intracranial disease. PMID- 9393196 TI - Endoscopic treatment of symptomatic spinal subarachnoid cysts. PMID- 9393197 TI - Normal lower limb venous Doppler flow phasicity: is it cardiac or respiratory? AB - OBJECTIVE: The purposes of this study were to determine the origin and nature of normal lower limb venous Doppler flow phasicity and to assess normal and respiratory variations. SUBJECTS AND METHODS: The common femoral veins of 12 healthy volunteers (three men and nine women; age range, 21-50 years; mean, 29 years) were evaluated by detailed spectral Doppler examinations with simultaneous ECG and respirometric tracings. The examinations were performed using a 5- or 7 MHz linear-array transducer with breath held in mid respiration, at the end of deep expiration, at the end of deep inspiration, during Valsalva's maneuver, and during quiet and deep breathing. The tracing obtained during breath-hold in mid respiration was considered the baseline. Tracings obtained during the other respiratory phases were analyzed for changes from the baseline. Doppler tracings were analyzed for phasicity, waveform frequency, components, velocities, velocity ratios, and presence of retrograde flow, all in correlation with simultaneous ECG and respirometric tracings. Tracings were analyzed independently by two observers to assess interobserver variability. RESULTS: With breath-hold in mid respiration, the common femoral vein Doppler tracings consisted of multiphasic waveforms that had a frequency similar to that of the heart rate. Each waveform consisted of systolic, v, diastolic, and a waves. The systolic wave occurred 0.4 sec later than the QRS complex of the ECG and was always antegrade. The v wave was always retrograde without flow reversal. The diastolic wave was always antegrade. The a wave was always retrograde but showed flow reversal in nine of 12 subjects. The systolic:diastolic velocity ratio ranged from 0.9 to 1.5 (mean, 1.1). The minimum:maximum velocity ratio ranged from -0.4 to 0.2 (mean, -0.15). With breath-hold at the end of expiration, the waveforms became slightly damped, becoming biphasic in five subjects and remaining multiphasic in seven. With breath-hold at the end of inspiration, the waveforms became nonphasic or biphasic in nine and decreased in velocity in 12. With Valsalva's maneuver, flow stopped. With normal respiration, cardiac waveforms were modulated by higher amplitude and less frequent biphasic respiratory waves. The plasticity was equal in two, dominantly cardiac in six, and dominantly respiratory in four. Flow velocity increased with expiration and decreased with inspiration. With deep breathing, the respiratory waves further increased, while the cardiac ones decreased in amplitude. The latter continued to modulate the respiratory phasicity in 10 subjects. CONCLUSION: During quiet respiration, lower limb venous Doppler tracings consisted of both cardiac and respiratory waveforms. Although respiratory waveforms disappeared when patients held their breath, Doppler tracings continued to be multiphasic and cardiac. Therefore, cardiac phasicity in lower limb venous Doppler tracings does not necessarily indicate cardiac disease. Other respiratory phases can modulate this basic cardiac pattern. Decrease in or loss of phasicity in these waveforms does not always mean proximal obstruction, because it can be caused by respiratory factors. Finally, the presence of minimal cyclic retrograde flow that is 5 cm/sec or less does not necessarily indicate cardiac disease. PMID- 9393198 TI - Encephalopathy after transjugular intrahepatic portosystemic shunting: analysis of incidence and potential risk factors. AB - OBJECTIVE: Our purpose was to estimate the incidence of encephalopathy after transjugular intrahepatic portosystemic shunting (TIPS) related primarily to the diversion of portal vein blood flow and to identify periprocedural factors to predict patients at risk. MATERIALS AND METHODS: All patients who underwent TIPS with at least 1 month of clinical observation after the procedure were monitored for clinically evident encephalopathy. Other variables that could individually induce encephalopathy were retrospectively analyzed for interrelationships with spontaneous or worsened encephalopathy. RESULTS: Of the 150 patients, 68 (45%) suffered from encephalopathy after TIPS, but in only 33 (22%) was it new or worse than baseline measurements obtained before TIPS; in 18 of these 33 patients, an underlying medical cause was implicated. Fifteen (10%) of the 150 patients developed mental dysfunction, usually mild and well controlled, thought to be related only to TIPS and not to any underlying morbidity. Low portal vein pressures after TIPS were found to be interrelated with new or worsened spontaneous encephalopathy (p = .04). Like-wise, advanced age (> 59 years old) weakly corresponded to the development of encephalopathy after TIPS. CONCLUSION: TIPS causes an acceptably low rate of encephalopathy that is usually mild. No specific variables exist for predicting the development or progression of encephalopathy after TIPS. PMID- 9393199 TI - Percutaneous revision of excess length from an implanted long-term central venous access device. PMID- 9393200 TI - Coarctation of the aorta: collateral flow assessment with phase-contrast MR angiography. AB - OBJECTIVE: The purpose of this report is to describe a new use of MR imaging in coarctation of the aorta. The specific question addressed was how well collateral blood flow in intercostal arteries, as determined by phase-contrast MR angiography, correlated with findings during surgery or catheterization in patients with coarctation of the aorta. CONCLUSION: Phase-contrast MR angiography is an excellent technique for detecting the presence or absence of collateral blood flow in the intercostal arteries of patients with coarctation of the aorta. Knowing whether collateral blood flow is present in patients with narrowing of the juxtaductal aorta should help assess the clinical hemodynamic significance of the coarctation. PMID- 9393201 TI - Endovascular repair of posttraumatic thoracic pseudoaneurysm with a stent graft. PMID- 9393202 TI - Intrathoracic extension of retroperitoneal disease: is it purely lymphatic spread or do some potential interfascial pathways exist? PMID- 9393203 TI - Helical CT angiography in gastrointestinal bleeding. PMID- 9393204 TI - Needle characteristics related to headaches after myelograms. PMID- 9393205 TI - Color Doppler sonography of renal artery aneurysm. PMID- 9393206 TI - The importance of being where? Balancing the perspective on practice coverage. PMID- 9393207 TI - Langerhans' cell histiocytosis of the synovium. PMID- 9393208 TI - MR imaging and CT appearances of aggressive angiomyxoma. PMID- 9393209 TI - Rupture of the right atrium-superior vena cava junction from blunt thoracic trauma: helical CT diagnosis. PMID- 9393210 TI - Migration of translumbar inferior vena cava catheter into the right ureter. PMID- 9393211 TI - CT and MR imaging of nasoethmoid schwannoma with intracranial extension. PMID- 9393212 TI - Ion channels. PMID- 9393213 TI - The health of gypsies. PMID- 9393214 TI - The use or uselessness of annual public health reports. PMID- 9393215 TI - Screening for fragile X syndrome: a model for genetic disorders? PMID- 9393216 TI - Replacing the NHS market. PMID- 9393217 TI - GMC accused of prejudicing doctors' defence. PMID- 9393218 TI - Court confirms right to palliative treatment for mental distress. PMID- 9393219 TI - Cancer in the offspring of radiation workers: a record linkage study. AB - OBJECTIVES: To test the "Gardner hypothesis" that childhood leukaemia and non Hodgkin lymphoma can be caused by fathers' exposure to ionising radiation before the conception of the child, and, more generally, to investigate whether such radiation exposure of either parent is a cause of childhood cancer. DESIGN: Case control study. SETTING: Great Britain. SUBJECTS: 35,949 children diagnosed as having cancer, together with matched controls. MAIN OUTCOME MEASURES: Parental employment as radiation worker as defined by inclusion in the National Registry for Radiation Workers and being monitored for external radiation before conception of child; cumulative dose of external ionising radiation for various periods of employment before conception; dose during pregnancy. RESULTS: After cases studied by Gardner and colleagues were excluded, fathers of children with leukaemia or non-Hodgkin lymphoma were significantly more likely than fathers of controls to have been radiation workers (relative risk 1.77, 95% confidence interval 1.05 to 3.03) but there was no dose-response relation for any of the exposure periods studied; indeed, the association was greatest for those with doses below the level of detection. No increased risk was found for fathers with a lifetime preconception dose of 100 mSv or more, or with a dose in the 6 months before conception of 10 mSv or more. There was no increased risk for the group of other childhood cancers. Mothers' radiation work was associated with a significant increase of childhood cancer (relative risk 5.00, 1.42 to 26.94; based on 15 cases and 3 controls). Only four of the case mothers and no controls were radiation workers during pregnancy. CONCLUSIONS: These results do not support the hypothesis that paternal preconception irradiation is a cause of childhood leukaemia and non-Hodgkin lymphoma; the observed associations may be chance findings or results from exposure to infective or other agents. If there is any increased risk for the children of fathers who are radiation workers, it is small in absolute terms: in Britain the average risk by age 15 years is 6.5 per 10,000; our best estimate, using all available data, is that the increase is 5.4 per 10,000. For mothers, the numbers are too small for reliable estimates of the risk, if any, to be made. PMID- 9393220 TI - Birth weight of offspring and mortality in the Renfrew and Paisley study: prospective observational study. AB - OBJECTIVE: To investigate the association between birth weight of offspring and mortality among fathers and mothers in the west of Scotland. DESIGN: Prospective observational study. PARTICIPANTS: 794 married couples in Renfrew district of the west of Scotland. MAIN OUTCOME MEASURES: Mortality from all causes and from cardiovascular disease over 15 year follow up. RESULTS: Women who had heavier babies were taller, had higher body mass index and better lung function, and were less likely to be smokers than mothers of lighter babies. Fathers of heavier babies were taller and less likely to be smokers than fathers of lighter babies. Mortality was inversely related to offspring's birth weight for both mothers (relative rate for a 1 kg lower birth weight 1.82 (95% confidence interval 1.23 to 2.70)) and fathers (relative rate 1.35 (1.03 to 1.79)). For mortality from cardiovascular disease, inverse associations were seen for mothers (2.00 (1.18 to 3.33)) and fathers (1.52 (1.03 to 2.17)). Adjustment for blood pressure, plasma cholesterol, body mass index, height, social class, area based deprivation category, smoking, lung function, angina, bronchitis, and electrocardiographic evidence of ischaemia had little effect on these risk estimates, although levels of statistical significance were reduced. CONCLUSIONS: Birth weight of offspring was related inversely to mortality, from all causes and cardiovascular disease, in this cohort. The strength of this association was greater than would have been expected by the degree of concordance of birth weights across generations, but an extensive range of potential confounding factors could not account for the association. Mortality is therefore influenced by a factor related to birth weight that is transmissible across generations. PMID- 9393221 TI - Impact of new antiretroviral combination therapies in HIV infected patients in Switzerland: prospective multicentre study. Swiss HIV Cohort Study. AB - OBJECTIVES: To examine trends in disease progression and survival among patients enrolled in the Swiss HIV cohort study during 1988-96 and to assess the influence of new antiretroviral combination therapies. DESIGN: Prospective multicentre study, with follow up visits planned at six monthly intervals. SETTING: Seven HIV units at university centres and cantonal hospitals in Switzerland. PATIENTS: 3785 men (mean age 35.0 years) and 1391 women (30.3 years) infected with HIV. 2023 participants had a history of intravenous drug misuse; 1764 were men who had sex with men; 1261 were infected heterosexually; and 164 had other or unknown modes of transmission. 601 participants had had an AIDS defining illness. RESULTS: During more than 15,000 years of follow up, there were 1456 first AIDS defining diagnoses and 1903 deaths. Compared with those enrolled during 1988-90, the risk of progression to a first AIDS diagnosis was reduced by 18% (relative risk 0.82 (95% confidence interval 0.73 to 0.93)) among participants enrolled in 1991-2, by 23% (0.77 (0.65 to 0.91)) among those enrolled in 1993-4, and by 73% (0.27 (0.18 to 0.39)) among those enrolled in 1995-6. Mortality was reduced by 19% (0.81 (0.73 to 0.90)), 26% (0.74 (0.63 to 0.87)), and 62% (0.38 (0.25 to 0.97)) respectively. Compared with no antiretroviral treatment, the risk of an initial AIDS diagnosis after CD4 lymphocyte counts fell to < 200 cells x 10(6)/1 was reduced by 16% (0.84 (0.73 to 0.97)) with monotherapy, 24% (0.76 (0.63 to 0.91)) with dual therapy, and 42% (0.58 (0.37 to 0.92)) with triple therapy. Mortality was reduced by 23% (0.77 (0.68 to 0.88)), 31% (0.69 (0.60 to 0.80)), and 65% (0.35 (0.20 to 0.60)) respectively. CONCLUSIONS: The introduction of antiretroviral combination therapies outside the selected patient groups included in clinical trials has led to comparable reductions in disease progression and mortality. PMID- 9393222 TI - Helicobacter pylori infection and mortality from ischaemic heart disease: negative result from a large, prospective study. AB - OBJECTIVE: To determine whether there is an independent association between Helicobacter pylori infection of the stomach and ischaemic heart disease. DESIGN: Prospective study with measurement of IgG antibody titres specific to H pylori on stored serum samples from 648 men who died from ischaemic heart disease and 1296 age matched controls who did not (nested case-control design). SUBJECTS: 21,520 professional men aged 35-64 who attended the British United Provident Association (BUPA) medical centre in London between 1975 and 1982 for routine medical examination. MAIN OUTCOME MEASURE: Death from ischaemic heart disease. RESULTS: The odds of death from ischaemic heart disease in men with H pylori infection relative to that in men without infection was 1.06 (95% confidence interval 0.86 to 1.31). In a separate group of 206 people attending the centre, plasma fibrinogen was virtually the same in those who were positive for H pylori (2.62 g/l) and those who were negative (2.64 g/l). CONCLUSIONS: A study that by its size and design minimised both random error and socioeconomic bias found no relation between H pylori infection and ischaemic heart disease. The validity of the study was shown by its confirmation of the recognised association between H pylori infection and stomach cancer (odds ratio 4.0 (1.9 to 8.2); P < 0.001). Eradication of H pylori infection may greatly reduce the incidence of stomach cancer, one of the most common causes of death from cancer worldwide, but it cannot be expected to have any effect in preventing ischaemic heart disease. PMID- 9393223 TI - What investigations and procedures do patients in hospices want? Interview based survey of patients and their nurses. PMID- 9393224 TI - Aseptic meningitis associated with high dose immunoglobulin: case report. PMID- 9393225 TI - Dentists' agreement on treatment of asymptomatic impacted third molar teeth: interview study. PMID- 9393226 TI - Why are babies getting heavier? Comparison of Scottish births from 1980 to 1992. PMID- 9393227 TI - Prospective case-control study of role of infection in patients who reconsult after initial antibiotic treatment for lower respiratory tract infection in primary care. AB - OBJECTIVE: To assess direct and indirect evidence of active infection which may benefit from further antibiotics in adults who reconsult within 4 weeks of initial antibiotic management of acute lower respiratory tract infection in primary care. DESIGN: Observational study with a nested case-control group. SETTING: Two suburban general practices in Arnold, Nottingham, over 7 winter months. SUBJECTS: 367 adults aged 16 years and over fulfilling a definition of lower respiratory tract infection and treated with antibiotics. 74 (20%) patients who reconsulted within 4 weeks for the same symptoms and 82 "control" patients who did not were investigated in detail at fallow up. MAIN OUTCOME MEASURES: Direct and indirect evidence of active infection at the time of the reconsultation or the follow up visit with the research nurse for the controls. Investigations performed included sputum culture, pneumococcal antigen detection, serial serology for viral and atypical pathogens and C reactive protein, throat swabs for detecting viral and atypical pathogens by culture and polymerase chain reaction, and chest radiographs. RESULTS: Demographic and clinical features of the groups were similar. Two thirds of the 74 patients who reconsulted received another antibiotic because the general practitioner suspected continuing infection. Any evidence of infection warranting antibiotic treatment was uncommon at reconsultation. The findings for the two groups were similar for the occurrence of identified pathogens; chest x ray changes of infection (present in 13%); and C reactive protein concentrations, which had nearly all fallen towards normal. Only three patients in the reconsultation group had concentrations > or = 40 mg/l. Pathogens identified at follow up in the 156 patients in both groups included ampicillin sensitive bacteria in six. Atypical infections diagnosed in 27 (Chlamydia pneumoniae in 22) and viral infections in 54 had probably been present at the initial presentation. CONCLUSION: Our study suggests that active infection, which may benefit from further antibiotics, is uncommon in patients who reconsult after a lower respiratory tract infection, and a repeat antibiotic prescription should be the exception rather than the rule. Other factors, such as patients' perception of their illness, may be more important than disease and infection in their decision to reconsult. PMID- 9393228 TI - Influence of patients' expectations on antibiotic management of acute lower respiratory tract illness in general practice: questionnaire study. AB - OBJECTIVE: To assess patients' views and expectations when they consult their general practitioner with acute lower respiratory symptoms and the influence these have on management. DESIGN: General practitioners studied consecutive, previously well adults and recorded clinical data, the certainty regarding their prescribing decision, and the influence of non-clinical factors on that decision. Patients completed a questionnaire at home after the consultation. SETTING: 76 doctors from suburban, inner city, and rural practices. SUBJECTS: 1014 eligible patients entered; 787 (78%) returned the questionnaire. MAIN OUTCOME MEASURES: The views of the patient, the views of and antibiotic prescription by the doctor. RESULTS: Most patients thought that their symptoms were caused by an infection (662) and that antibiotics would help (656) and had both wanted (564) and expected (561) such a prescription. 146 requested an antibiotic, 587 received one. Of the 643 patients who thought they had an infection, 582 wanted an antibiotic and thought it would help. Severity of symptoms did not relate to wanting antibiotics. For those prescribed antibiotics, their doctor thought they were definitely indicated in only 116 cases and not indicated in 126. Patient pressure most commonly influenced the decision to prescribe even when the doctor thought antibiotics were not indicated. Doctors considered antibiotics definitely indicated in only 1% of the group in whom patient pressure influenced the prescribing decision. Patients who did not receive an antibiotic that they wanted were much more likely to express dissatisfaction. Dissatisfied patients reconsulted for the same symptoms twice as often as satisfied patients. CONCLUSION: Patients presenting with acute lower respiratory symptoms often believe that infection is the problem and antibiotics the answer. Patients' expectations have a significant influence on prescribing, even when their doctor judges that antibiotics are not indicated. PMID- 9393229 TI - Recent advances. Care of near term infants with respiratory failure. PMID- 9393230 TI - ABC of palliative care. Anorexia, cachexia, and nutrition. PMID- 9393231 TI - Case finding for the fragile X syndrome and its consequences. PMID- 9393232 TI - Life span: conception to adolescence. PMID- 9393233 TI - The real ethics of rationing. Purchasers, not surgeons, control waiting lists. PMID- 9393234 TI - The real ethics of rationing. Doctors should not be penalised for doing private work in spare time. PMID- 9393235 TI - The real ethics of rationing. Conflict between private and national interests within NHS needs scrutinising. PMID- 9393236 TI - Death rates from childhood leukaemia near nuclear sites. Numbers of observed deaths were closer to those expected after known risk factors were allowed for. PMID- 9393237 TI - Death rates from childhood leukaemia near nuclear sites. Other studies showed that radiation levels in Newbury area were low. PMID- 9393238 TI - Death rates from childhood leukaemia near nuclear sites. Epidemiological studies must define their cohort objectively. PMID- 9393239 TI - Death rates from childhood leukaemia near nuclear sites. Findings were probably due to chance fluctuations in small numbers of deaths. PMID- 9393240 TI - Death rates from childhood leukaemia near nuclear sites. Place of residence at diagnosis and at death may be different. PMID- 9393241 TI - Death rates from childhood leukaemia near nuclear sites. Number of deaths in Newbury area is not increased. PMID- 9393242 TI - In screening for breast cancer, clinical examination is as effective as mammography. PMID- 9393243 TI - Incidence of gastrointestinal side effects due to alendronate is high in clinical practice. PMID- 9393244 TI - Communication between GPs and cooperatives is poor for terminally ill patients. PMID- 9393245 TI - Geriatric depression scale can be used in older people in primary care. PMID- 9393246 TI - Is the clinical course of HIV infection changing? Finding is disheartening. PMID- 9393247 TI - Is the clinical course of HIV infection changing? Exclusion of people not followed up for 12 months may have biased results. PMID- 9393248 TI - Is the clinical course of HIV infection changing? Study's censoring strategy may be source of bias. PMID- 9393249 TI - Through the looking glass: backwards. PMID- 9393250 TI - Therapeutic modification of nuclear factor kappa B binding activity and tumor necrosis factor-alpha gene expression during acute biliary pancreatitis. AB - The role of cytokines has been well documented in the pathogenesis of acute pancreatitis. Antibodies against specific cytokines have been used to treat pancreatitis, with mixed results. The transcription factor nuclear factor (NF) kappa B is a pleiotropic regulator of many genes involved in stress and inflammatory responses. The aim of this study was to prevent the NF-kappa B binding activity and tumor necrosis factor (TNF)-alpha gene overexpression as a possible therapeutic intervention for acute pancreatitis. Reversible acute biliary pancreatitis was induced in male Sprague Dawley rats as established in this laboratory. The animals were sacrificed at 0, 5, 15, 30 min and 1, 2, 3, 4, 6, 8, 10, 12, and 24 hours after the induction of pancreatitis. NF-kappa B binding activity was determined by electrophoretic mobility shift assay, and TNF alpha gene expression was assayed by reverse transcription-PCR. NF-kappa B binding activity was markedly higher around 4 hours and persisted up to 24 hours after pancreatitis induction in animals with acute pancreatitis, whereas TNF alpha mRNA levels peaked at 24 hours. When amobarbital (to block NF-kappa B activation) was given (60 mg/kg body weight, I.P.) 3 hours before induction of pancreatitis, the activation of NF-kappa B and the overexpression of TNF-alpha gene was prevented, with significantly decreased severity of pancreatitis as assessed by amylase and clinical recovery. We conclude that 1) preventing the activation of NF-kappa B eliminates the induced overexpression of inflammatory cytokines (TNF-alpha) in acute pancreatitis, 2) such intervention correlates with clinical improvement in pancreatitis, and 3) this genetic modification offers a possible therapeutic intervention in acute pancreatitis. PMID- 9393251 TI - Small molecule inhibition of tumor necrosis factor gene processing during acute pancreatitis prevents cytokine cascade progression and attenuates pancreatitis severity. AB - The morbidity and mortality associated with acute pancreatitis are primarily a result of pancreatic parenchymal necrosis and the development of marked pulmonary dysfunction. Recent evidence suggests that both of these conditions are propagated by interleukin (IL)-1 beta and tumor necrosis factor (TNF)-alpha, which are produced in large quantities within these organs. Because the generation of these cytokines occurs in a predictable manner early in the development of acute pancreatitis, we aimed to determine whether cytokine gene processing could be inhibited in vivo and what effects this would have on pancreatitis severity. Mild [caerulein, 50 micrograms/kg/hour intraperitoneally (IP) x 4; n = 40] or severe (choline-deficient diet; n = 40) necrotizing pancreatitis was induced in NIH swiss mice. Animals were randomly given a novel small molecule (CNI-1493; 10 mg/kg IP) known to inhibit macrophage production of TNF and IL-1 in vitro by inhibiting translation of TNF mRNA into protein. Control animals received IP vehicle. All animals with acute pancreatitis showed dramatic up-regulation of the IL-1 beta and TNF-alpha genes. Those animals receiving CNI 1493 demonstrated attenuated production of both species of mRNA in pancreatic as well as pulmonary tissue (P < 0.01). Markers of pancreatitis severity such as serum amylase and lipase, as well as pancreatic necrosis, were decreased in animals treated with CNI-1493 (all P < 0.05). Posttranscriptional blockade of TNF production precludes induction of the proinflammatory cytokine cascade that normally occurs during acute pancreatitis. This lack of cytokine gene processing in the pancreas and lungs results in dramatic reductions in tissue damage and pancreatitis severity, which is not model dependent. This is the first time that a small molecule has been shown to influence this disease. PMID- 9393252 TI - Prospective evaluation of selective lymph node biopsy for cutaneous malignant melanoma. AB - Some patients presenting with cutaneous malignant melanoma without palpable adenopathy have regional metastatic disease. We have applied the technique of gamma probe-directed selective lymph node biopsy and used the results to direct further therapy. The results of a prospective nonrandomized clinical study are presented. Between November 1993 and December 1996, 63 patients with a diagnosis of primary cutaneous malignant melanoma underwent lymphoscintigraphy with technetium sulfur colloid followed by gamma probe-guided lymph node biopsy. There were 32 (51%) women and 31 (49%) men with a mean age of 51.1 years (median, 50; range, 13-87). Mean Breslow thickness was 2.13 mm (range, 0.5-15.0 mm; median, 1.56 mm). Primary locations were head and neck in 8 (13%), trunk in 24 (38%), upper extremity in 13 (21%), and lower extremity in 18 (29%). Selective lymph node biopsy was done on an outpatient basis with local anesthesia in 49 cases (78%) and in the operating room with general anesthetic in 14 patients (22%). One lymph node site was biopsied in 46 patients (73%), two sites in 16 (25%), and three in 1 (2%), for a total of 81 selective lymph node biopsy sites, mean 1.29 per patient. The mean number of labeled lymph nodes removed per site per patient was 1.64 (range, 1-5). Seroma or infection occurred in 6 patients (10%). Micrometastatic disease was identified in nine selective lymph node biopsy sites in eight patients. Of eight patients undergoing lymph node dissection, 5 (63%) had no additional pathological lymph node involvement. With a mean follow-up of 579 days from selective lymph node biopsy (median, 594; range, 36-1157), 59 (94%) have no evidence of disease. Three patients have died, 2 with systemic disease (475 and 1149 days) and 1 from a myocardial infarction (380 days). No patient has failed with regional-only recurrence. Gamma probe-directed selective lymph node biopsy is a straightforward procedure that can be done in the outpatient setting and facilitates management of patients with cutaneous malignant melanoma. PMID- 9393253 TI - Quality assessment of intraoperative blood salvage and autotransfusion. AB - Intraoperative blood salvage and autotransfusion are commonly used to minimize exposure to banked blood. Although this technique has been used widely for years, data vary regarding the quality of autotransfused blood. Salvaged blood may contain plasma, residual heparin, and free hemoglobin released from damaged cells. All of these factors may contribute to the adverse sequelae sometimes seen with autotransfusion. For these reasons, we have monitored autotransfused blood to assess its quality. Intraoperative blood salvage was used during most cardiac procedures and at the discretion of the surgeon in other specialties. Blood was collected through a double lumen catheter that was anticoagulated with heparin, filtered, centrifuged, and washed with saline. A sample of the blood was removed for analysis, which included hematocrit, heparin assay, fibrinogen, and free hemoglobin levels. Over a 6-year period, 1593 patients had intraoperative blood salvage with quality assessment. The majority of patients underwent cardiac operations (941 patients, 59%), whereas 243 had orthopedic (15%) and 208 had vascular (13%) procedures. Additionally, there were 127 pediatric patients (8%) and 74 miscellaneous procedures (5%). The highest average yield of salvaged blood was during vascular procedures (1073 +/- 76 mL), whereas orthopedic cases had the lowest yield (378 +/- 19 mL) and hematocrit (39%). There was minimal residual heparin activity, even in patients requiring systemic anticoagulation (0.3 to 0.5 units/mL). Patients undergoing pediatric procedures had the lowest concentration of free hemoglobin (476 mg/L), whereas all adult patients had higher free hemoglobin levels, especially vascular operations (990 mg/L). Intraoperative salvaged blood has minimal heparin activity, even in procedures requiring systemic anticoagulation. Fibrinogen, a marker of residual plasma, was undetectable in the majority of cases. These data indicate that intraoperative blood salvage generally results in a high-quality product (good hematocrit, low heparin, minimal plasma), although there are significant differences in free hemoglobin levels depending on the operative procedure. PMID- 9393254 TI - Mesh inguinal herniorrhaphy: a ten-year review. AB - Inguinal herniorrhaphy remains one of the most common surgical operations, with approximately 10 to 20 per cent performed for recurrence. Reviews by specialized hernia centers show mesh repair has a recurrence rate of 0.2 per cent. Detractors of this repair include increased cost, technical difficulty, and risk for infection. The purpose of this study was to compare mesh versus nonmesh inguinal herniorrhaphy at a large teaching institution. From 1985 to 1994, 892 patients underwent primary repair for inguinal hernia at the Veterans Administration Hospital at Memphis, TN. Patients were stratified by repair [Lichtenstein (Mesh), open anterior (Bassini, Marcy, McVay, and Shouldice), laparoscopic (Lap), and preperitoneal (Post)]. Operative time for Mesh repair (111 +/- 2 minutes) was longer than for Bassini or McVay (91 +/- 2 and 98 +/- 2 minutes; P < 0.05), and Lap repairs were longer than all others (192 +/- 16 minutes; P < 0.05). Hospital stay averaged 2.2 +/- 0.1 days for Mesh versus 2.6 +/- 0.1 days for all repairs combined (P = not significant). Mesh patients developed four wound infections (1.0%), none requiring mesh removal, versus nine infections (1.8%) in other groups (P = not significant). One Mesh patient (0.3%) developed recurrence, compared with 16 (3.5%) with open anterior repair (P < 0.01). Inguinal herniorrhaphy using an open mesh repair technique provides superior recurrence rates without increasing risk for infection, length of stay, or technical difficulty. PMID- 9393255 TI - Image-guided core biopsy has advantages over needle localization biopsy for the diagnosis of nonpalpable breast cancer. AB - Image-guided core biopsy (IGCB) of nonpalpable mammographic abnormalities has gained attention as an alternative to needle-localized breast biopsy (NLB). This study evaluated IGCB in the diagnostic workup of patients with nonpalpable mammographic lesions suspicious for cancer. Eighty-six patients who underwent IGCB were compared to 85 patients who underwent NLB for the diagnosis of mammographic lesions suspicious for cancer. The incidence of positive margins was less in patients who subsequently underwent needle-localized resection in the IGCB group than in the NLB group (29 and 65%; P < 0.0001). The volume of excision was greater for patients in the IGCB group than for the NLB group (106 cm3 and 52 cm3; P < 0.0001). Patients in the IGCB group averaged 1.1 operative procedures compared with patients in the NLB group, who required an average of 1.9 operative procedures. The mean charge for an IGCB was $1011 compared to $2975 for a NLB. Subset analysis of 32 spiculated masses from the IGCB group and 21 from the NLB group showed similar advantages of IGCB over NLB. The preoperative use of IGCB for mammographically suspicious lesions can reduce the incidence of positive surgical margins and the number of surgical procedures required. The use of IGCB allows for a more efficient diagnostic workup and less expense to the patient. PMID- 9393256 TI - Ductal carcinoma in situ of the breast: correlation of pathologic and mammographic features with extent of disease. AB - Optimal treatment of ductal carcinoma in situ (DCIS) of the breast requires an improved understanding of its pathologic extent and propensity for local recurrence. This study was performed to analyze mammographic and pathologic features of DCIS that might predict the extent of disease within the breast and facilitate treatment selection between lumpectomy alone, lumpectomy and radiotherapy, and mastectomy. At our institution, 60 cases of DCIS were diagnosed in 59 patients from June 1985 to February 1995 and form the basis of this retrospective study. Demographic and treatment-related information was obtained from hospital and tumor registry records. Mammograms were reviewed and size estimates of the abnormalities were determined. Pathologic slides from all cases were reviewed and classified according to size group, focality, nuclear grade, necrosis, and histologic subtype. DNA ploidy status and proliferation indices were available for 28 patients. Pathologically, 43 (72%) cases were < 15 mm, 14 (23%) were 16 to 40 mm, and 3 (5%) were > 40 mm. Five (8%) of the lesions were multicentric, 28 (47%) focal, and 27 (45%) multifocal. Thirty-three (55%) patients were treated by mastectomy, 16 (27%) by lumpectomy alone, and 11 (18%) by lumpectomy and radiation therapy. Mammographic size, histologic grade, presence or absence of necrosis, histologic subtype, DNA ploidy, and proliferative index were compared with pathologic size and focality by chi 2 analysis. Mammographic size correlated significantly with pathologic size (chi 2 = 11.3; P = 0.02) but underestimated the extent of disease in 9 cases. Although focality correlated significantly with pathologic size (chi 2 = 15.8; P = 0.003), the remaining histopathologic features did not significantly correlate with pathologic size or focality. Histopathologic features, including DNA studies, do not reliably predict the pathologic extent of DCIS, but mammographic size and focality do significantly correlate with pathologic size. Nevertheless, most cases of DCIS are small focal or multifocal lesions that are amenable to breast conservation approaches; further studies are needed to determine the appropriate use of lumpectomy, radiation therapy, and mastectomy in the treatment of DCIS. PMID- 9393257 TI - Utility of fine-needle aspiration cytology and frozen-section examination in the operative management of thyroid nodules. AB - Fine-needle aspiration cytology has a high sensitivity for the diagnosis of solitary thyroid nodules. Certain diagnoses involving follicular histologies often cannot be made with needle biopsy alone. The utility of frozen-section examination of thyroid nodules, with particular regard to those lesions with follicular histologies, is also limited. We examined the correlation of fine needle aspiration cytology and frozen-section examination in solitary thyroid nodules to determine the contribution of frozen-section examination to the operation. We reviewed the fine-needle aspiration cytology, frozen-section examination, and final pathology of 100 consecutive patients undergoing thyroidectomy for a solitary solid thyroid nodule in an 4-year period. The diagnoses were classified as indeterminant, benign, or malignant. The utility and impact of the diagnosis from fine-needle aspiration or frozen section on the operative procedure performed was analyzed. Fine-needle aspiration cytology as a diagnostic test for thyroid nodules demonstrated an indeterminant rate of 23 per cent, with a diagnostic accuracy of 77 and 92 per cent for benign and malignant disease, respectively. In all patients with inaccurate benign diagnosis on fine needle aspiration cytology, follicular neoplasm was misinterpreted for follicular adenoma or multinodular goiter. In comparing frozen-section results, the indeterminant, benign, and malignant rates were 7, 96, and 64 per cent, respectively. Of the 23 patients with indeterminant results on fine-needle aspiration cytology, the intraoperative frozen-section diagnosis on 4 patients was deferred to permanent section; 18 received accurate cytological diagnosis; and in 1 patient, carcinoma was missed. Overall, the decision about the extent of surgical thyroid resection was changed in only 2 patients based on the frozen section results. Preoperative evaluation with fine-needle aspiration cytology can accurately and appropriately define the extent of thyroid surgery in most patients with a diagnosis of malignant neoplasm or benign disease. Intraoperative frozen-section examination may be helpful if fine-needle aspiration cytology results are inderminant and in cases of follicular histology as an adjunct for evaluation of the thyroid nodule, but overall, frozen section does not contribute to the management of the thyroid lesion at the time of surgery. PMID- 9393258 TI - Delayed ulcer recurrence after gastric resection: a new postgastrectomy syndrome? AB - Recurrent ulceration following gastrectomy for peptic ulcer disease typically occurs within the first several years postoperatively. Since 1990, we have managed 20 patients who had undergone previous gastrectomy for peptic ulcer and developed ulcer recurrence more than 10 years postoperatively. Mean age at recurrence was 64 years, and the average time from original surgery to recurrent ulceration was 21 years (range, 10-36 years). All patients had undergone vagotomy and antrectomy (17 patients) or subtotal gastrectomy (3 patients). Presenting symptoms included gastric outlet obstruction (70%) and bezoar formation (60%). Endoscopic findings in this group of patients included a stenotic gastric outlet (70%) and marginal ulcerations (80%). Thirteen of 15 patients tested (87%) were Helicobacter pylori positive. Reoperation included partial resection of the gastric pouch and exploration for persistent vagus nerve. Twelve patients underwent Roux-en-Y reconstruction, whereas eight patients had Bilroth II reconstruction. Three of the latter group also had Braun enteroenterostomy performed. Good to excellent clinical results were obtained in 80 per cent of patients. The four patients with poor outcomes shared the following characteristics: 1) H. pylori-positive status, 2) presence of a preoperative bezoar, 3) Roux-en-Y reconstruction. Our current approach is to avoid Roux-en-Y construction in favor of Braun enteroenterostomy. Further prospective analysis of long-term postgastrectomy patients is needed to determine whether this clinical picture represents a new postgastrectomy syndrome. PMID- 9393259 TI - Cost utility of routine imaging with Tc-99m-sestamibi in primary hyperparathyroidism before initial surgery. AB - Tc-99m-sestamibi has been shown to localize parathyroid adenomas effectively, but controversy continues as to the use of this scan before initial surgery for primary hyperparathyroidism. We analyzed the cost utility of obtaining this study before initial surgery for primary hyperparathyroidism. Twenty-two consecutive patients with primary hyperparathyroidism underwent dual-phase Tc-99m-sestamibi scan before initial bilateral neck exploration. Surgical findings were correlated with the results of sestamibi scan. There were 15 women and 7 men, with a mean age of 50.5 years (range, 22-76). Preoperative mean total calcium was 11.74 mg/dL (range, 10-15), ionized calcium was 6.19 mg/dL (range, 5.2-7.7), and intact parathyroid hormone was 153.5 pg/mL (range, 83.1-551). Postoperative mean ionized calcium was 4.56 mg/dL (range, 4.1-5.57). Twenty sestamibi scans had a positive localization, and 2 scans had no localization. At surgery, 18 solitary adenomas, 3 diffuse hyperplasias, and 1 patient with four normal parathyroid glands were found. Sixteen sestamibi scans were true positive (solitary adenoma), 4 scans were false positive (2 diffuse hyperplasia, 1 wrong side, and 1 lymph node), 1 negative scan was true negative (diffuse hyperplasia), and 1 negative scan was false negative (adenoma). One patient (four normal glands) at the second operation had a supernumerary fifth gland adenoma excised from the mediastinum. Preoperative Tc-99m-sestamibi scan did not offer any advantage when a complete bilateral neck exploration is performed. Sixteen of (84%) adenomas were correctly localized, but 18 of 19 adenomas were in the neck and were easily found. The 1 ectopic adenoma was not found by scanning or with initial surgery. The 4 of 22 (18%) false-positive localizations and the 2 of 22 (9%) negative scans contributed nothing to the surgery. Of the 22 localizing sestamibi scans, surgery was not altered to affect the outcome. At a cost of $550 per sestamibi scan and with the error inherent in the scan, it is not cost effective to obtain Tc-99m sestamibi scan before initial surgery for primary hyperparathyroidism. PMID- 9393260 TI - Vacuum pack technique of temporary abdominal closure: a four-year experience. AB - The purpose of this review is to present a 4-year experience with the vacuum pack technique of temporary abdominal closure. From April 1992 to December 1996, 171 vacuum packs were performed on 93 patients. Eighty-seven vacuum packs were performed on 38 general surgical patients, and 84 vacuum packs were performed on 55 trauma patients. Overall hospital mortality was 32 per cent. Methods of achieving permanent wound closure varied in 73 patients. Four patients (4.3%) developed enterocutaneous fistulae; four patients developed intra-abdominal abscesses (4.3%). There were no eviscerations. Management of the complicated intra-abdominal process is discussed: 1) the decision to manage the abdomen in an open fashion; 2) which method of temporary closure to use; 3) subsequent explorations; 4) when the abdomen should be closed; 5) which type of closure to use; and 6) when the abdominal wall should be revised (herniorrhaphy). The vacuum pack is the method of choice for open abdomen management and temporary abdominal closure at our institution. With careful subsequent management, good patient outcome can be achieved. PMID- 9393261 TI - Prolonged abdominal packing for trauma is associated with increased morbidity and mortality. AB - Abdominal packing and planned reoperation is a lifesaving technique for temporary control of hemorrhage in severely injured patients. Morbidity and mortality in this group of patients, however, remain significant. It is unclear whether the duration of packing impacts upon outcome. The purpose of this study is to evaluate the abscess, sepsis, and mortality rates associated with duration of abdominal packing. The records of 35 patients treated with abdominal packing between July 1994 and December 1995 who survived to reoperation were retrospectively reviewed. Evaluation included age; sex; mechanism; injuries; Abdominal Trauma Index; duration of packing; survival; and presence of abscess, sepsis or other infections. Patients packed for a total of 72 hours or less had lower abscess, sepsis, and mortality rates than those packed for more than 72 hours. The differences in abscess rate and mortality were statistically significant (P < 0.05). The Abdominal Trauma Index and mechanism of injury were similar for the two groups. Based on these results, we conclude that although abdominal packing is a useful technique in the severely injured patient, it is associated with greater morbidity and mortality when the duration of packing exceeds 72 hours. PMID- 9393262 TI - Nonoperative therapy for acute necrotizing pancreatitis. AB - Acute necrotizing pancreatitis is a highly morbid and lethal condition. We performed a retrospective study of all patients admitted to Louisiana State University Medical Center between 1980 and 1995 with a diagnosis of pancreatitis (N = 617) and specifically examined those (N = 26) who developed acute necrotizing pancreatitis. During the period 1980 to 1989, there were 7 patients who progressed to acute necrotizing pancreatitis. Six of these seven patients died (mortality, 86%). These patients were managed with multiple operations for debridement and necrosectomy. The age ranged from 31 to 86 years in this group, with a mean of 58.5. The patients' total hospital days ranged from 2 to 125 days with a mean of 63.5 days. In 1989, we adopted an initial nonoperative approach to necrotizing pancreatitis and began using CT-guided catheter drainage for this condition. During this time period, 19 patients have progressed to necrotizing pancreatitis. The range of hospital days was from 13 to 90 days, with a mean of 43.8 days. There were 2 deaths in this last group, resulting in a mortality rate of 10.5 per cent. All of these patients were treated nonoperatively in the acute phase of their illness. Two patients (15.8%) subsequently underwent laparotomy and drainage when the collections were not amenable to CT-guided drainage. Morbidity in this population approached 70 per cent; however, the mortality was only 10 per cent compared to 86 per cent in the previous group. Although nonoperative therapy has its associated morbidity, and although we understand the controversy surrounding the management of this condition, it appears at least in this population to have much less mortality than those who were treated operatively in the acute phase. PMID- 9393263 TI - Surgical management of chronic pain from chronic pancreatitis. AB - Chronic pain from chronic pancreatitis remains a difficult clinical problem. We present the results of surgical attempts to control this pain. For the past 3 years, all patients with chronic pancreatitis and pain requiring high-dose narcotics or hospitalization for pain control were evaluated by the following algorithm. Any anatomic pathology causing ductal dilatation was surgically addressed first (Puestow's procedure, pseudocyst drainage, or sphincteroplasty). If there was no evidence of ductal dilatation, or if pain recurred postoperatively, denervation procedures were performed (splenopancreatic flap, thorascopic sympathectomy, or resection). Pain recurrence was defined as the need for further hospitalization or reoperation. Data were analyzed by comparison of two proportions. Follow-up averaged 26 months. Thirty-seven patients underwent 44 operations solely in an attempt to control pain; 62 per cent were male, and 70 per cent had chronic alcoholic pancreatitis. Our results show that surgical management provides relief in 68 per cent of patients, and no one procedure is clearly superior to others. PMID- 9393264 TI - Stereotactic core biopsy of the breast: results of one-year follow-up of 101 patients. AB - Stereotactic core biopsy (SCB) is being used as a cost-effective alternative to needle localized biopsy (NLB). However, an area of concern is the potential for sampling error, with sparse surgical data available concerning follow-up and failure rates. We therefore reviewed our results in patients undergoing SCB for mammographically detected breast abnormalities. Between January 1994 and February 1995, 128 patients underwent SCB. Average age was 56.4 years. Nine patients (7.0%) had histologic evidence of malignancy, with 111 (86.7%) benign diagnoses requiring no further initial intervention. Eight patients (6.3%) proceeded directly to NLB, five because of technical failure of SCB and three because of suspicious initial histology. One of the latter patients had ductal carcinoma in situ. The remaining 111 SCB patients were evaluated at 6 months and 1 year by mammographic and physical examination. Ten patients were lost to follow-up. Of the remaining 101 patients, 98 (97%) had stable mammograms and normal physical examinations. Three patients (3.9%) required subsequent NLB due to progression of the mammographic lesion. Two cases were histologically benign, and 1 patient had ductal carcinoma in situ adjacent to the previous SCB biopsy site. An additional patient underwent NLB for a new radiographic abnormality at a separate location in the ipsilateral breast, which was invasive ductal carcinoma. SCB appears to be an effective alternative to NLB for the majority of patients deemed eligible. Careful mammographic follow-up is warranted for these patients given the small, but real, possibility of sampling error. PMID- 9393266 TI - Parotid pleomorphic adenoma. PMID- 9393265 TI - Helicobacter pylori and peptic ulcer surgery. PMID- 9393267 TI - Preventing postoperative recurrence of Crohn's disease. AB - BACKGROUND AND METHODS: Risk factors for recurrence of Crohn's disease and the evidence for progress in reducing recurrence following resection were reviewed. A Medline based literature review was carried out. RESULTS AND CONCLUSION: Only smoking has been confirmed as a significant adverse risk factor for recurrence. Evidence for differing recurrence rates in fibrostenosing disease and perforating disease is inconclusive, but such a classification along with the endoscopic findings of recurrence may have a place in the analysis of therapeutic trials. Minimal resectional surgery with clearing of only macroscopic disease seems to be justified, with no clear benefits from different anastomotic techniques. Recent trials offer encouraging evidence of the usefulness of 5-aminosalicylic acid, particularly higher-dose regimens started early after resection, although the long-term benefits are uncertain. The oral steroid, budesonide, offers a potent treatment with minimal side-effects, but evidence of its prevention of recurrence is presently weak. PMID- 9393268 TI - Rational approach to combined carotid and ischaemic heart disease. AB - BACKGROUND: The management of patients with concomitant coronary and carotid artery disease remains a controversial subject. The aim of this review was to develop a rational plan for the management of such patients based on a review of the literature. METHOD AND RESULTS: A retrospective review was carried out of relevant papers derived from the Medline database from 1964 to 1996. CONCLUSION: The management of patients with concomitant coronary and carotid artery disease has not yet been put to the test in a properly designed and randomized multicentre trial. It is suggested that, until the results of such a trial are available, the rational approach to combined symptomatic disease is combined carotid endarterectomy and coronary artery bypass grafting (CABG). Combined surgery is also appropriate for patients with symptomatic carotid artery disease and significant but asymptomatic cardiac disease. At present there is inadequate evidence to promote carotid endarterectomy for asymptomatic disease in combination with CABG. PMID- 9393269 TI - Monoclonal antibody treatment of colorectal cancer. AB - BACKGROUND: The recent development of adjuvant monoclonal antibody immunotherapy for patients suffering from colorectal cancer has led to a re-evaluation of the role of these molecules in the treatment of solid tumours. In particular, interest has been directed at identifying appropriate candidates for therapy, evaluating treatment schedules and developing new molecules of therapeutic potential. METHODS: This is a review of published data on patients undergoing antibody therapy. In addition, current theories of the mechanism of action of antitumour monoclonal antibodies are presented, along with potential future therapeutic approaches. RESULTS AND CONCLUSION: Monoclonal antibody-based adjuvant therapy of colorectal cancer appears to be effective; international multicentre trials continue. The development of new molecules, such as chimaeric antibodies, offers the potential of increased tumour targeting with reduced toxicity. Such molecules may be used alone or in combination with agents such as chemotherapy or cytokines. PMID- 9393270 TI - Long-term oral administration of branched chain amino acids after curative resection of hepatocellular carcinoma: a prospective randomized trial. The San-in Group of Liver Surgery. AB - BACKGROUND: Live cirrhosis is a state of protein calorie malnutrition which may induce various complications. This study aimed to elucidate if long-term nutritional support with branched chain amino acids (BCAAs) is effective after hepatic resection for hepatocellular carcinoma (HCC). METHODS: Between 2 and 3 weeks after operation, 75 patients were randomized to receive oral BCAAs (Aminoleban EN) 100 g per day for 1 year, and another 75 patients were assigned to a control group. Mean follow-up times were 35.8 months and 36.0 months respectively. RESULTS: Flapping tremor was less common, body-weight was greater and performance status was better in the BCAA-treated group throughout the 1-year period. BCAA treatment significantly increased red blood cell and serum albumin level in patients with Child grade B and C disease. Substantially similar effects were observed in those with major hepatic resection. Higher Fischer molar ratios were maintained in the treated group. However, there were no differences in cumulative tumour recurrence and survival rates. CONCLUSION: Long-term oral nutritional support with BCAAs after resection of HCC is beneficial in improving clinical features and laboratory data without increasing the rate of tumour recurrence, particularly in patients with advanced cirrhosis or after major hepatic resection. PMID- 9393271 TI - Prognostic scoring systems to predict outcome in peritonitis and intra-abdominal sepsis. AB - BACKGROUND: Early classification of patients presenting with peritonitis and intra-abdominal sepsis by means of objective scoring systems is desirable to select patients for 'aggressive' surgery and to compare results of different treatment regimens. However, none of the existing scoring systems has fulfilled all expectations. METHODS: Evaluation of the value of various scoring systems (Acute Physiology And Chronic Health Evaluation (APACHE) II, Simplified Acute Physiology Score, Sepsis Severity Score, Multiple Organ Failure, Mannheim Peritonitis Index (MPI), Ranson and Imrie) was performed in 50 patients. Additionally, scoring systems were combined to obtain a 'combined score' for the prediction of peritonitis-related in-hospital death. Hazard ratios were calculated in a univariate and multivariate analysis. RESULTS: In the univariate analysis all scoring systems, except Ranson and Imrie, predicted the primary outcome. In the multivariate analysis, only the APACHE II score (hazard ratio 6.7) and the MPI (hazard ratio 9.8) contributed independently to the prediction of outcome. All patients with an APACHE II score of 20 or more and a MPI of 27 or more died in hospital. CONCLUSION: Combination of the APACHE II and the MPI provides the best scoring system fitting clinical goals. PMID- 9393272 TI - Randomized controlled trial of prophylactic chest physiotherapy in major abdominal surgery. AB - INTRODUCTION: This randomized controlled study evaluated the clinical benefit and physiological effects of prophylactic chest physiotherapy in open major abdominal surgery. METHODS: A group of 174 patients received chest physiotherapy including breathing with pursed lips, huffing and coughing, and information about the importance of early mobilization. In addition high-risk patients were given resistance training on inspiration and expiration with a mask. The resistance used during inspiration was -5 cmH2O and that during expiration +10 cmH2O. The control group (194 patients) received no information or treatment unless a pulmonary complication occurred. RESULTS: Oxygen saturation on postoperative days 1-3 was significantly greater in the treatment group. Treated patients were mobilized significantly earlier. No difference was noted in peak expiratory flow rate or forced vital capacity. Postoperative pulmonary complications occurred in 6 per cent of patients in the treatment group and in 27 per cent of controls (P < 0.001). In high-risk patients the numbers with pulmonary complications were six of 40 and 20 of 39 respectively. Pulmonary complications were particularly common in patients with morbid obesity. CONCLUSION: Preoperative chest physiotherapy reduced the incidence of postoperative pulmonary complications and improved mobilization and oxygen saturation after major abdominal surgery. PMID- 9393273 TI - Acquired immune deficiency syndrome-related intussusception in adults. PMID- 9393274 TI - Hand-held Doppler as a screening test in primary varicose veins. AB - BACKGROUND: Hand-held Doppler is in common use for evaluating varicose veins, but its accuracy in identifying the exact sites of venous reflux is inferior to that of duplex scanning. It has been suggested that duplex should be used to investigate all varicose veins, but this is currently impractical, and should be unnecessary if hand-held Doppler examination were shown to be an adequate screening test. METHODS: Eighty-five patients (122 legs) with primary varicose veins were evaluated using a hand-held Doppler in the outpatient clinic, according to a protocol. Patients then had venous duplex imaging. RESULTS: Different methods of assessing the long saphenous vein (LSV) (tourniquet and tapping tests, and examination at and below the groin) had similar sensitivities for detecting reflux (75-86 per cent), and together detected 91 per cent of cases. Six of the nine missed had a competent saphenofemoral junction, and five had low-velocity reflux. Hand-held Doppler assessment missed 11 cases of popliteal fossa reflux; only four involved the short saphenous vein (SSV), and most had low-velocity popliteal vein reflux. CONCLUSION: Hand-held Doppler examination missed LSV or SSV incompetence in 11 per cent of legs, but these included cases with short-duration and low-velocity reflux of dubious clinical importance. PMID- 9393275 TI - Lateral rectal ligaments contain important nerves. PMID- 9393276 TI - Population-based study of the treatment and prognosis of carcinoma of the rectum. AB - BACKGROUND: Few population-based studies address the issue of treatment of carcinoma of the rectum (15 cm or less from the anal verge) both from surgical and epidemiological aspects. METHODS: Some 827 patients were analysed in the cancer registry of the Cote-d'Or (Burgundy, France) from 1976 to 1990 (493,931 inhabitants). RESULTS: Resection for cure increased from 57.2 per cent before 1981 to 77.0 per cent after 1985 (P < 0.001), and the proportion of Dukes A and B cases from 35.8 to 52.5 per cent (P < 0.001). Among patients resected for cure, continence-preserving resections were performed more frequently during the 1986 1990 period (48.0 per cent) than during the two previous 5-year periods (20.0 per cent; P < 0.001), more often in women, in the upper half of the ampulla and for tumours of less than 45 mm. The operative mortality rate after curative surgery decreased from 13.9 to 3.7 per cent (P < 0.001) between the first and the last period whereas the 5-year crude survival rate rose from 25.8 to 42.6 per cent (P < 0.001). Age, stage of disease and period of diagnosis were independent prognostic factors of death in a relative survival model. CONCLUSION: This study indicates that significant advances have been achieved at a population level in the treatment of rectal cancer in terms of diagnosis, continence-preserving procedures and survival. PMID- 9393277 TI - Bone assessment in patients with ileal pouch-anal anastomosis for inflammatory bowel disease. AB - BACKGROUND: Patients with ulcerative colitis are at risk of low bone mineral density (BMD). Proctocolectomy with ileal pouch-anal anastomosis (IPAA) for ulcerative colitis diminishes the risk of bone disease. The aims of this study were to assess the mechanism of low BMD and to measure bone density changes after IPAA. METHODS: Twenty patients with IPAA for ulcerative colitis, of mean(s.d.) age 38(9) (range 21-58) years, had measurements of lumbar spine and femoral neck BMD by dual energy X-ray absorptiometry, a mean(s.d.) 28(23) (range 3-84) months after proctocolectomy. Serum levels of calcium, phosphate, parathyroid hormone, osteocalcin and 25-hydroxy vitamin D were determined. Fifteen patients were followed for 28(12) (range 8-50) months. RESULTS: At baseline, six patients had spine BMD more than two standard deviations below the normal value, and three had vertebral crush fractures. Mean vitamin D values were normal and no patient had osteomalacia. BMD increased with time elapsed since IPAA (spine: r = 0.71, P = 0.005). During follow-up, mean(s.d.) changes in bone density were +2.3(3.8) and +2.1(5.6) per cent per year at the spine and femoral neck respectively. CONCLUSION: These results suggest that in patients with IPAA for ulcerative colitis, low BMD is not associated with vitamin D malabsorption and may be reversible after surgery. PMID- 9393278 TI - Magnetic resonance imaging of the pelvic floor in patients with obstructed defaecation. AB - BACKGROUND: Evacuation proctography and measurements of anorectal physiology are frequently used to clarify the pathophysiology of obstructed defaecation. In some patients these tests are normal, despite convincing clinical evidence of defaecatory difficulty. The aim of this study was to determine whether magnetic resonance imaging (MRI) could reveal pelvic floor abnormality in patients with obstructed defaecation. METHODS: Eleven women with obstructed defaecation, in whom evacuation proctography and anorectal physiology were normal, were examined by MRI, using a fast gradient echo sequence. Measurements of pelvic visceral and muscular descent were taken at rest and during straining, and compared with those obtained from 13 asymptomatic volunteers. RESULTS: Patients with obstructed defaecation had significantly greater pelvic visceral descent (P < 0.05), levator muscle descent (P = 0.04), levator plate angle change (P = 0.003) and increase in the area of the pelvic floor hiatus (P = 0.0002) than asymptomatic volunteers. CONCLUSION: MRI demonstrated marked pelvic visceral and levator muscle descent in women with obstructed defaecation, despite normal evacuation proctography and anorectal physiology. MRI should be considered if these examinations have been normal. PMID- 9393279 TI - Tumour bed positivity predicts outcome after breast-conserving surgery. AB - BACKGROUND: Local recurrence after breast-conserving surgery is associated with a short distant disease-free survival, particularly if it occurs early. Early recurrence is caused by residual disease left at the time of surgery. Previous studies have demonstrated that disease in the tumour bed is a common finding after breast-conserving surgery. METHODS: The follow-up (mean 4.4 years) of 300 patients who had tumour bed analysis performed by the cavity shaving technique following breast-conserving surgery is presented. Postoperative radiotherapy was administered to all patients. RESULTS: The incidence of tumour bed positivity was 39.3 per cent. With a selective re-excision policy the local recurrence rate was 2.0 per cent and distant recurrence rate 10.4 per cent. Multivariate analysis identified lymphovascular invasion, oestrogen receptor status and tumour bed status as independent predictors of time to distant recurrence. CONCLUSION: A low rate of local recurrence can be achieved using this technique of margin assessment. Tumour bed status may be a useful prognostic factor following breast conserving surgery. PMID- 9393280 TI - Pathological appearance of the stomach after endoscopic mucosal resection for early gastric cancer. AB - BACKGROUND: The pathological findings of the resected stomach after endoscopic mucosal resection (EMR) for early gastric cancer were reviewed. EMR was indicated when a lesion consisting of well or moderately differentiated adenocarcinoma had a diameter of less than 2 cm. METHODS: Of 39 patients with early gastric cancer were treated with EMR between 1990 and 1995, 11 required additional surgery. RESULTS: Malignant tissue in the gastric wall was completely removed in four patients, while cancer cells remained in the mucosa adjacent to the scar in five and infiltrated into the submucosa in two. Most of these residual cancers were characterized by a lesion with a diameter exceeding 15 mm and by the location in the body or cardia of the stomach. Lymph node metastases were observed in one patient whose carcinoma invaded the deeper submucosal layer. Assessment of the depth of entire invasion from the endoscopically-resected specimen was correct for six of 11 patients. CONCLUSION: Gastric carcinomas to be resected by EMR should be smaller, especially if located in the body or cardia. Accurate diagnosis of the width and depth of invasion is indispensable before proceeding to EMR. Surgery may be the treatment of choice when there is submucosal invasion. PMID- 9393282 TI - Laparoscopic fenestration of symptomatic non-parasitic cysts of the liver. PMID- 9393283 TI - Colour flow duplex imaging of occlusive arterial disease of the lower limb. PMID- 9393281 TI - Risk factors for surgical treatment in the Dutch Gastric Cancer Trial. AB - BACKGROUND: A multicentre randomized study of surgical treatment of gastric cancer has shown increased mortality and morbidity rates in patients having D2 resection. The aim of this report is to analyse risk factors in these patients. METHODS: In a prospective randomized trial, comparing two types of lymphadenectomy for curable gastric cancer, risk factors for hospital death and morbidity in 711 patients treated with curative intent were evaluated by multivariate analysis using stepwise regression analysis. RESULTS: Age greater than 65 years and male sex were the most important risk factors for death (relative risk (RR) 4.35 (95 per cent confidence interval (c.i.) 2.07-9.15) and 2.51 (95 per cent c.i. 1.24-5.08) respectively). The extent of nodal dissection was also a significant risk factor for death (RR 2.13). For overall complications, splenectomy was the most important risk factor (RR 2.13 (95 per cent c.i. 1.44-3.16)), while pancreatectomy and type of gastrectomy were the only factors significantly influencing the occurrence of major surgical complications. CONCLUSION: The cumulative mortality risks of these factors should be considered carefully when planning surgery for individual patients. PMID- 9393284 TI - Intragastric endoscopic surgery using the transanal endoscopic microsurgery technique. PMID- 9393285 TI - One-wound laparoscopic cholecystectomy. PMID- 9393286 TI - One-wound laparoscopic cholecystectomy. PMID- 9393287 TI - One-wound laparoscopic cholecystectomy. PMID- 9393288 TI - Antisperm antibodies and male subfertility. AB - The outcome of treatment for couples with male immunological infertility must be considered in conjunction with the risks, costs and time to success involved. Reversible factors should be treated. Steroid treatment is a good option but its failure in two-thirds of patients should add impetus to attempts to predict responders before or soon after starting treatment. The treatment options are not mutually exclusive [48]. The success of IUI may be enhanced further by new sperm preparation techniques. If severe immunological infertility exists or other infertility factors are present, IVF or ICSI may be considered from the outset. PMID- 9393289 TI - Prognostic implications of cytogenetic findings in kidney cancer. AB - OBJECTIVE: To evaluate the prognostic impact of cytogenetic findings in renal cell carcinoma (RCC). PATIENTS AND METHODS: Tumour cytogenetics, histomorphology, DNA ploidy and S-phase fraction, stage, size, and grade were related to survival in 50 consecutive patients with RCC. The mean follow-up was 3.9 years (median 4.2, range 0-8.8). RESULTS: The predictive probability for recurrence-free survival at 5 years (5-year RFS) for all 50 patients was 0.54. There was a significant association between the degree of cytogenetic complexity and survival, in that patients with five or less aberrations had a better prognosis than those with more than five changes (5-year RFS 0.71 and 0.43, respectively; P < 0.05). Patients with del(8p)/-8, +12, and +20 had a significantly worse prognosis compared with those without these aberrations. Of the well-known prognostic variables grade and stage, the former was far better for predicting prognosis. A Spearman correlation test showed a significant covariation of grade with the S-phase fraction, T-stage, and cytogenetic complexity. CONCLUSION: The degree of cytogenetic complexity and recurrent cytogenetic abnormalities affect the prognosis in RCC, although grade was the most reliable independent prognostic factor predicting disease recurrence. PMID- 9393290 TI - Clinical review of 100 consecutive surgically treated patients with upper urinary tract transitional tumours. AB - OBJECTIVE: To determine the outcome of conservative or radical treatment in a retrospective study of 100 consecutive patients with upper urinary tract tumours. PATIENTS AND METHODS: From 1965 to 1995, 100 patients (78 men and 22 women, mean age 65 years, range 27-82) with upper urinary tract tumours were treated surgically, using nephroureterectomy with excision of a cuff of bladder in 53 and organ-sparing treatment in 47. The outcome was assessed as survival and recurrence during a follow-up of up to 15 years. RESULTS: After radical and organ sparing treatment, the 15-year cancer-specific survival was 69% and 25%, respectively; metastases developed in 17% and 19% and global recurrence in 40% and 70%, respectively. While locoregional and bladder recurrences were similar in the two groups (9% vs 8% and 30% vs 38%, respectively), ureteric-stump recurrence in the conservative group was 23%. There were no significant differences in survival rates between patients with single or multiple presentation, or for localization, while the grading of the lesions proved to be an accurate prognostic indicator. CONCLUSION: This experience of urothelial neoplasia of the upper tract highlights the difficulty in diagnosing this pathology and in entrusting screening to a non-invasive technique such as urinary cytology. The percentage recurrence observed after organ-sparing therapy indicates that this treatment should be used cautiously. PMID- 9393291 TI - Using the ICSOoL to measure the impact of lower urinary tract symptoms on quality of life: evidence from the ICS-'BPH' Study. International Continence Society- Benign Prostatic Hyperplasia. AB - OBJECTIVE: To present and describe the validity and reliability of the International Continence Society-Benign Prostatic Hyperplasia study quality-of life (ICSQoL) instrument, a new set of questions to assess the impact of lower urinary tract symptoms (LUTS) on quality of life (QoL) in middle-aged and elderly men. PATIENTS AND METHODS: The study comprised 1271 consecutive men over the age of 45 years, attending urology departments in 12 countries, with LUTS and possible benign prostatic obstruction who were recruited to the ICS-'BPH' study (the clinic group); 423 ambulant men were recruited from a general practice in the UK to provide a community group. Each individual completed the ICS-'BPH' study questionnaire which includes six items addressing general and specific aspects of QoL (the ICSQoL). Content and construct validity were assessed by interviews with patients and by testing hypotheses within the study groups, e.g. the relationships with age, individual LUTS (as measured on the ICSmale questionnaire) and generic health status, as measured by the Short Form (SF-36) and EuroQol instruments. Reliability was assessed by measures of internal consistency and a test-retest analysis. RESULTS: The ICSQoL items were easily understood by patients, were completed with low levels of missing data, and address some (but not all) concerns about the impact of LUTS on QoL. The ICSQoL items have good construct validity, showing expected differences between community and clinic samples, and expected relationships with each other and individual LUTS. Items had good test-retest reliability, but their internal consistency was poor, confirming that ICSQoL questions should not be combined into a score. General ICSQoL items were closely related with most domains of the SF-36 and the EuroQol. CONCLUSION: ICSQoL items may be used individually or as a group in research studies or in clinical practice. PMID- 9393292 TI - Construction and validation of a short-form benign prostatic hypertrophy health related quality-of-life questionnaire. BPH Group in General Practice. AB - OBJECTIVE: To construct and validate a short-form benign prostatic hypertrophy (BPH) health-related quality-of-life (HRQL) questionnaire which is more practical in use and as informative as the 20-item visual analogue scale questionnaire (QOL20) previously validated in French. PATIENTS AND METHODS: From the factorial structure of the QOL20, a nine-item questionnaire (QOL9) was constructed using stepwise linear regression and factorial analysis. The feasibility and reliability of the QOL9 were analysed in a cross-sectional case-control study and a longitudinal cohort study, including symptomatic patients with BPH treated for 6 months with an alpha 1-blocker (alfuzosin). RESULTS: The reduction of the QOL20 to QOL9 showed a minimal loss of information (90-95% of the variance of QOL20 was explained by QOL9) and lead to a three-dimensional structure: well being, patients' perceived sexual-life status, and BPH interference with activities. The QOL9 was practical in use (completion rate 87-100%; duration of completion at inclusion 11.6, SD 2.0 min), consistent (Cronbach's alpha > 0.7), reliable (intraclass correlation coefficient > 0.80) and responsive (effect-size index 0.9, SD 0.01 in the longitudinal study). CONCLUSIONS: The QOL9 is a good BPH HRQL questionnaire, including an assessment of patients' perceived sexual-life status; it easy to administer, accurate, reproducible and responsive to change with treatment. We suggest that the QOL9 be substituted for the QOL20 and administered in addition to the International Prostate Symptom Score to obtain a better assessment of the patients' perception of their disease. PMID- 9393293 TI - The influence of colonic enema irrigation on urodynamic findings in patients with neurogenic bladder dysfunction. AB - OBJECTIVE: To evaluate whether colonic enema irrigation influences the urodynamic characteristics of patients with spina bifida, an overactive bladder and detrusor sphincter dyssynergia (DSD). PATIENTS AND METHODS: Since 1991, 83 patients with spina bifida at our institution have treated their bowel dysfunction by colonic irrigation every 24-48 h. In 12 patients (seven boys and five girls, mean age 7.7 years, range 0.7-13.8) with an overactive bladder and DSD, urodynamic studies of the bladder before and after enema treatment were available with no intercurrent changes in urological therapy. RESULTS: There were no significant changes overall in bladder capacity, leak-point pressure, bladder compliance and bladder instability in the selected group of children. CONCLUSION: Although enema therapy for bowel treatment in patients with spina bifida gave good results for faecal incontinence, with good patient compliance, no favourable effect on bladder function should be expected in most patients with a high-risk urinary tract dysfunction. Further study is needed to determine factors in patients who will benefit urologically from enema treatment. PMID- 9393294 TI - Acute suppression of idiopathic detrusor instability with magnetic stimulation of the sacral nerve roots. AB - OBJECTIVE: To assess the effect of magnetic stimulation of the S3 nerve root on unstable contractions in patients with idiopathic detrusor instability. PATIENTS AND METHODS: Twelve patients with idiopathic instability were studied. The S3 nerve root was localized by mapping the response of the toe flexor muscles and anal sphincter to magnetic stimulation at different sites. Unstable contractions were provoked by rapidly infusing saline into the bladder and the effect of magnetic stimulation of S3 on contractions was assessed. RESULTS: Magnetic stimulation relieved the sensation of urinary urgency and reduced the duration and amplitude of provoked contractions in all patients. Stimulation reduced the area under the pressure/time curve by 80-98%. In some patients there was a shortlived residual suppressive effect lasting up to 90 s. CONCLUSIONS: Magnetic stimulation of S3 acutely abolishes unstable contractions in patients with idiopathic detrusor instability. PMID- 9393295 TI - Kinetics of peptide-induced release of inflammatory mediators by the urinary bladder. AB - OBJECTIVE: To investigate the release of inflammatory mediators by the urinary bladder in response to exposure to pro-inflammatory peptides. MATERIALS AND METHODS: Isolated guinea pig urinary bladder was incubated with 10 mumol/L each of substance P (SP), neurokinin A (NKA), calcitonin gene-related peptide (CGRP), vasoactive intestinal peptide (VIP), octreotide acetate (a long-acting analogue of somatostatin, SOM), or bradykinin (BK), and the release of histamine, prostaglandin (PG) E2, PGF2 alpha and leukotriene B4 (LTB4) was determined during 0-5, 5-30 and 30-120 min after addition. RESULTS: Substance P, NKA, VIP and BK stimulated the release of histamine, while CGRP and SOM suppressed the release to below the spontaneous rates. All peptides, except CGRP and SOM, stimulated the release of PGE2 between 0 and 30 min, and only VIP failed to stimulate the release of PGF2 alpha within 5 min of exposure. Substance P, NKA, VIP and BK stimulated the release of LTB4 and this required > 5 min of exposure. CONCLUSION: These results indicate that the peptides evaluated induce an immediate and transient release of histamine and activation of cyclooxygenase and delayed activation of 5-lipoxygenase. These actions may directly regulate the participation of these peptides in the pathogenesis of cystitis. PMID- 9393296 TI - Management of the long-term urinary catheter in the asymptomatic patient in the Accident and Emergency department. AB - OBJECTIVE: To determine the management of long-term urinary catheters in the asymptomatic patient in the Accident and Emergency (A&E) Department. PATIENTS AND METHODS: Using data obtained from patient records, a retrospective study was undertaken of 41 patients who presented to the A&E department of a large district general hospital, on a total of 80 occasions in a 6-month period, with blocked or bypassing long-term urinary catheters, but who were otherwise asymptomatic. RESULTS: In 78% of presentations, patients had one or more investigations performed on their urine, which in 15% was on urine aspirated through an old, unchanged catheter. In 23% of presentations, patients were discharged only having had their catheters flushed, and in only 41% was the catheter changed as the first line of management. Finally, in 63% of episodes, patients were discharged with antibiotics, despite having no symptoms of a urinary tract infection. CONCLUSIONS: All patients with long-term urinary catheters will have bacteriuria and performing investigations on this urine if the patient is asymptomatic is a waste of resources. Long-term urinary catheters which become blocked usually do so by encrustation on the luminal surface of the catheter. Flushing these catheters may dislodge some of these encrustations, but ideally the catheter should be changed. The prescribing of antibiotics to asymptomatic patients with bacteriuria has no proven benefit, but on the contrary it may cause harmful side effects and also selects for antibiotic-resistant bacteria. These patients should be managed in the A&E department with a simple catheter change. PMID- 9393297 TI - Preliminary experience with a urinary control device in the management of women with genuine stress incontinence. AB - OBJECTIVE: To evaluate the efficacy and acceptability of the FemAssist urinary control device (Insight Medical UK Ltd). PATIENTS AND METHODS: Twenty-seven women with cystometrically confirmed genuine stress incontinence performed a perineal 1 h pad-test; the pad test was then repeated with the FemAssist device in place and the difference in pad weights compared with and without the device in place. Patients were given 100 mm visual analogue scales (VAS) to measure discomfort, acceptability and embarrassment associated with using the device. RESULTS: The median (range) loss with and without the FemAssist device was 4.9 (0-65) mL and 21 (1-94), respectively (P < 0.01); 20 patients were less wet when using the device. The median (range) VAS scores were; discomfort 35 (4-93), embarrassment 11 (0-75), and acceptability 65 (3-100). Discomfort was greater among the women with a greater loss. The acceptability correlated negatively with discomfort (r = -0.53) and negatively with embarrassment (r = -0.39); 15 patients (56%) reported that they would use the device in the long-term. CONCLUSION: The FemAssist device produced a significant reduction in urine loss. The magnitude of benefit could not be predicted for an individual and the device was ineffective in some women. The use of the device was influenced by discomfort and associated embarrassment. The device has a role in the management of stress incontinence but patients must be assessed individually for suitability. PMID- 9393298 TI - Collagen for female genuine stress incontinence after a minimum 2-year follow-up. AB - OBJECTIVE: To evaluate the medium-term outcome of gluteraldehyde cross-linked (GAX) collagen in the treatment of genuine stress incontinence in women. PATIENTS AND METHODS: The study comprised 111 women (age range 33-90 years) with genuine stress incontinence who were treated with para-urethral collagen injections between 1990 and 1995. The patients were followed prospectively and their clinical outcome documented. Pre- and post-operative urodynamic data were examined to determine any prognostic indicators. RESULTS: The overall results at a minimum 2-year follow-up (mean 3.3) showed 25% of patients to be dry and a further 40% improved. Although there were significant changes in some urodynamic values, no predictive factors of success were identified. Previous surgery for stress incontinence did not influence the final outcome. CONCLUSION: Para urethral collagen injection is a safe and relatively simple procedure with acceptable results at the medium-term follow-up. It could be offered as a primary or secondary procedure to women with genuine stress incontinence who are unable or unwilling to undergo surgical treatment. Objective urodynamic assessment revealed no factors of prognostic significance. PMID- 9393299 TI - Tumour progression and survival in patients with T1G3 bladder tumours: 15-year outcome. AB - OBJECTIVE: To evaluate tumour progression and survival of patients with T1G3 bladder tumours who were followed for 15 years. PATIENTS AND METHODS: A subset of 48 patients with T1G3 bladder tumours was entered into a randomized trial of transurethral resection (TUR) or TUR plus bacille Calmette-Guerin (BCG) therapy and followed for a minimum of 15 years. Thirty-nine (81%) patients received one or more courses of BCG. The endpoints of the study were stage progression (defined as muscle invasion of metastasis) and disease-specific survival. RESULTS: Of the 48 patients, 25 (52%) progressed and 15 (31%) died from the disease; 33 patients (69%) survived, including 24 (50%) with an intact bladder. The median progression-free survival time was 151 months. Tumour progression occurred in 35% of the patients within the first 5 years, in 16% after 5-10 years and in 12% of those followed for 10-15 years. Deaths from cancer occurred in 25% of the patients in the first 5 years and in 10% of patients at risk from 5 to 15 years. CONCLUSIONS: Patients with T1G3 bladder tumours who are treated conservatively are at life-long risk of having a muscle-invasive tumour and dying from bladder cancer. PMID- 9393300 TI - Soluble urinary CD14 after intravesical bacille Calmette-Guerin immunotherapy for carcinoma in situ. AB - OBJECTIVE: To characterize the production of the monocyte activation marker, soluble CD14 (sCD14), after bacille Calmette-Guerin (BCG) immunotherapy for superficial bladder cancer. PATIENTS AND METHODS: Nineteen patients with carcinoma in situ were treated with a standard regimen of intravesical BCG. Urine samples were collected after each instillation and analysed; the levels of soluble CD14 were determined by an enzyme-linked immunosorbent assay, the molecular weight confirmed by Western blotting and the possible cell source investigated using immunohistochemistry. RESULTS: The mean levels of sCD14 were higher in patients with persistent carcinoma (designated as failures) than in those who had successful complete responses (responders) to BCG immunotherapy. The differences were statistically significant, with P = 0.034 at instillation 4 and P = 0.027 at instillation 5 for the total mass of sCD14, and P = 0.049 at instillation 4 for the concentration of sCD14. The predominant type of sCD14 in urine was the 48 kDa form, although in most patients there was a minor band of reactivity at 54 kDa. A panel of human bladder cancer cell lines did not react with the anti-CD14 monoclonal antibodies 3C10, 5C5 and BA8, and the antibodies also failed to react with malignant epithelial cells in frozen sections of untreated bladder tumour. Furthermore, sCD14 was not secreted by cultured bladder tumour cells. The source of urinary sCD14 is likely to be the resident and infiltrating macrophages in the bladder wall. Freshly isolated peripheral blood monocytes secreted sCD14 in response to BCG in a manner analogous to stimulation with bacterial lipopolysaccharide. CONCLUSION: A soluble form of CD14 is secreted into the urine of patients who receive intravesical BCG. The measurement of soluble urinary CD14 could be of prognostic significance for the response to immunotherapy. PMID- 9393301 TI - Effect of oral administration of prostaglandin E1 on erectile dysfunction. AB - OBJECTIVES: To investigate the effect of limaprost, an oral prostaglandin E1 (PGE1) derivative, on erectile dysfunction and to compare the effects of limaprost with a Chinese herbal drug, gosyajinki-gan. PATIENTS AND METHODS: The study comprised 50 consecutive patients with mild erectile dysfunction who showed a good erectile response to intracavernosal injection with 20 micrograms of PGE1. Limaprost was administered to the first 25 patients (30 micrograms three times daily) and gosyajinki-gan (7.5 g three times daily) to the next 25 patients, for 8 consecutive weeks. Patients were evaluated by their ability to achieve vaginal penetration and by a subjective assessment of erectile function (penile rigidity and maintenance of erection) before and after the treatment, using a self administered questionnaire. Objective measurements (nocturnal penile tumescence, NPT, values) were also evaluated. RESULTS: Eleven of the 24 patients who received limaprost without interruption and four of the 24 taking gosyajinki-gan succeeded in vaginal penetration; the difference in the positive response rate was significant. The mean increment of NPT was significantly higher with limaprost treatment. However, all positive responders in both groups did not experience a full erection. There were no side-effects in any patient except one in the limaprost group who developed a facial flush. Thus the treatment was mild enough to be tolerated. CONCLUSION: Limaprost was more effective for mild erectile dysfunction than was gosyajinki-gan. PMID- 9393302 TI - Variability in penile appearance and penile findings: a prospective study. AB - OBJECTIVE: To document prospectively variation in penile morphology and clinical findings in children. PATIENTS AND METHODS: The study comprised a consecutive sample of 468 boys whose consultation with a physician included a genital examination in a primary-care paediatric practice in rural northern Wisconsin. RESULTS: Circumcised boys under 3 years of age were significantly more likely to have a partially or completely covered glans, a reddened meatus, balanitis, or trapped epithelial debris, and less likely to have a fully exposed glans than were circumcised boys of 3 years or older. Among the 238 boys under 3 years, those circumcised were significantly more likely to have non-cosmetic problems, including coronal adhesions, trapped epithelial debris, a reddened meatus, preputial stenosis (phimosis) and balanitis, than were boys with a foreskin. Findings in the circumcised group under 3 years included: fully exposed glans (n = 78, 35.6%), partially covered glans (n = 67, 30.6%), adhesions (25.6%), completely covered glans (20.1%), entrapped desquamated epithelial debris (24.7%), reddened meatus (19.1%), balanitis (15.5%), and preputial stenosis (0.9%). Only two genital examinations in boys with foreskins revealed pertinent findings. Coronal adhesions develop in circumcised boys at 2-6 months of age and usually resolve by 24 months. The degree of skin covering the glans after neonatal circumcision peaks at 6 months of age. CONCLUSIONS: There are significant variations of appearance in circumcised boys; clinical findings are much more common in these boys than previously reported in retrospective studies. The circumcised penis requires more care than the intact penis during the first 3 years of life. Parents should be instructed to retract and clean any skin covering the glans in circumcised boys, to prevent adhesions forming and debris from accumulating. Penile inflammation (balanitis) may be more common in circumcised boys; preputial stenosis (phimosis) affects circumcised and intact boys with equal frequency. The revision of circumcision for purely cosmetic reasons should be discouraged on both medical and ethical grounds. PMID- 9393303 TI - Histological evidence of decreased contralateral testicular blood flow during ipsilateral testicular torsion. AB - OBJECTIVE: To evaluate contralateral testicular blood flow by histological examination of arterioles during ipsilateral testicular torsion. MATERIALS AND METHODS: The study comprised two groups of 20 male albino rats (weight 250-270 g). The control group underwent a sham operation, while in the second group the left testes were twisted clockwise by 720 degrees. All rats underwent bilateral orchidectomy 24 h after the initial intervention. Three slides for each testis (n = 240) were evaluated randomly while unaware of treatment to determine the total number of arterioles, the percentage of collapsed and open arterioles, the diameter of open arterioles and the presence or absence of blood cells in the lumen. Differences were assessed using t-tests for paired and independent samples. RESULTS: Very few arterioles were collapsed in both testes of the control group and in the ipsilateral testes of the torsion group, but half the arterioles in the contralateral testes of the torsion group were collapsed. The difference between the contralateral testes of the two groups and between the ipsilateral and contralateral testes in the torsion group were significant. The diameter of uncollapsed arterioles did not differ significantly among either testes of the control and torsion groups and either testes in each group. Both testes in the torsion group had significantly more arterioles containing blood cells than those in the control group. The difference between the testes in the torsion group was also significant, but was not in the control group. CONCLUSION: There was histological evidence of decreased blood flow in the contralateral testis during unilateral testicular torsion; contralateral testicular damage during unilateral testicular torsion may result from hypoxia caused by decreased blood flow. PMID- 9393304 TI - The agreement among urological experts on the diagnostic management of patients with common urological problems. AB - OBJECTIVE: To assess the level of agreement among randomly selected international urologists on the diagnostic management of patients with prostate cancer, bladder cancer, urinary stones or lower urinary tract symptoms (LUTS) arising from benign prostatic hyperplasia (BPH). METHODS: A computer program was used to provide an unbiased format of 53 simulated patients, comprising 13 with prostate cancer, 10 with bladder cancer, 10 with stones in the upper urinary tract and 20 with LUTS from BPH. For each case, the history was provided to the user while information from 60 diagnostic tests could be chosen interactively. Thirty-three university based urologists participated in the study. The probability that a certain test was used by them in a certain patient [P(test)] and the related costs (Swedish 1995 prices) were recorded. The probability that two urologists would agree (relative measure of agreement, RMA) on whether or not to use one particular test in a certain case was RMA(test) = P(test)2 + [1-P(test)]2 and the mean of this RMA(test) for a certain patient [RMA(case)] was used as a measure of the inter individual agreement among the urologists on the diagnostic management. The significance levels of the generalized kappa statistic, KG, were also calculated. The correlation between the RMA(case) and the diagnostic groups was analysed. RESULTS: The KG was statistically significant for all cases; the RMA(case) was significantly correlated with the diagnostic groups (rs = 0.86). The agreement in the diagnostic management was the strongest for stones, then for bladder cancer and prostate cancer, and the weakest for BPH. The mean cost for the diagnostic evaluation for one case varied from $455 to $1771 (mean 898) and varied in the diagnostic groups, i.e. $1718 for prostate cancer, $947 for bladder cancer, $400 for stones and $594 for BPH. CONCLUSION: The diagnostic management of urological patients varies greatly among urological experts from the industrial world. As a consequence, the related diagnostic costs might vary by about 400% if prices were similar everywhere. The agreement on the diagnostic management of cases is strongly correlated to the diagnosis. LUTS from BPH seems to be managed with the poorest agreement. PMID- 9393305 TI - The natural history of pelvi-ureteric junction obstruction in children presenting clinically with the complaint. AB - OBJECTIVE: To determine the natural history of untreated pelvi-ureteric junction (PUJ) obstruction in children presenting clinically with the complaint. PATIENTS AND METHODS: The study comprised 42 children with anatomical PUJ obstruction (three with bilateral lesions) who were managed expectantly in the first instance, who had no immediately troublesome symptoms and differential function in the affected kidney(s) of > 40%. They were followed by serial ultrasonography and dynamic diuresis renography. Pyeloplasty was advised for those with persistent symptoms or where differential renal function fell below 40%. RESULTS: Only 15 children had presented with clearly relevant symptoms (loin pain or febrile urinary infection). Thirty-four kidneys showed obstructive renographic curves initially and 38 had moderate or severe hydronephrosis. During the follow up (range 14-120 months, mean 56) patients remained asymptomatic except for four of the nine presenting with loin pain. Renographic curves were apt to change, with 'obstruction' increasing or decreasing with time, in the latter instance usually with lessening or resolution of hydronephrosis. Eleven patients underwent pyeloplasty, five for symptoms and six because of deteriorating renal function. Renal function did not decline in any patient with mild hydronephrosis and/or a non-obstructive renographic curve at presentation, but did so disproportionately in those with severe hydronephrosis or a Type IV renographic curve. CONCLUSIONS: Patients effectively asymptomatic at presentation are likely to remain so. Expectant management is appropriate for patients with mild hydronephrosis and/or non-obstructive renographic curves at the outset. Conversely, pyeloplasty may be advisable for those with severe hydronephrosis or Type IV renographic curves. Otherwise, for most the natural history of the complaint remains to be determined. PMID- 9393306 TI - Paediatric ureteroscopy for ureteric calculi: a 4-year experience. AB - OBJECTIVE: To study the efficacy and safety of ureteroscopy in the management of paediatric ureteric calculi at various levels. PATIENTS AND METHODS: The records of 50 ureteroscopic procedures performed on 43 children (age range 6 months-12 years) for primary ureteric calculi or ureteric fragments after electrohydraulic shockwave lithotripsy (EHWL) were analysed retrospectively. The distribution of the calculi was in nine in the upper upper, seven in the mid and 30 in the lower ureter, with a mean stone burden of 1.26 cm. Ureteroscopy was performed with the 8.5/9.5/11.5 F Wolf ureteroscopes, and EHL was used as the primary method of fragmentation. Intravenous urograms were available in 34 children (79%) and micturating cystography (MCUG) was performed in 23 children (54%) during the follow-up. RESULTS: An overall stone-free status was achieved in 40 children (93%) after performing 47 ureteroscopies (94%). Stones were completely cleared in 78%, 100% and 97% of the procedures in the upper, mid and lower ureters, respectively. In three children the procedure failed and they were salvaged by uretero-lithotomy. During the procedures, an upper ureteric perforation occurred in one patient and a lower perforation in another. MCUG revealed low grade vesico ureteric reflux in 17% of the patients, but close follow-up showed the reflux to be sterile and clinically insignificant. CONCLUSION: Ureteroscopy was safe and effective for the management of mid and lower ureteric calculi but the results for upper ureteric calculi were marginally inferior. Ureteroscopy must be performed judiciously to minimize ureteric injury in children. The incidence of vesico-ureteric reflux after mechanical dilatation of the intramural ureter was infrequent and clinically insignificant. PMID- 9393307 TI - The use of a multipurpose stent in children. AB - OBJECTIVES: To assess the use of a multipurpose stent (the 'Blue stent', Angiomed Urosoft Pyeloplasty Stent, Bard, UK) in children undergoing pyeloplasty and ureteric reimplantation. PATIENTS AND METHODS: Between August 1994 and August 1996, the Blue stent was used in 50 renal units in 46 children aged 2 months to 12 years and 6 months. Twenty-five children underwent pyeloplasty, 11 had ureteric reimplantation for vesico-ureteric reflux (VUR), eight had ureteric reimplantation with remodelling for obstructed megaureters and in two patients it was used during the removal of stones. The mean follow-up was 18 months (range 6 30 months). RESULTS: After pyeloplasty, 22 patients (88%) had improved renal function and drainage with a decrease in hydronephrosis; two patients had a decrease in hydronephrosis only, one had an anastomotic leak and needed a repeat pyeloplasty and four developed a urinary tract infection (UTI). After ureteric reimplantation, VUR was not detected in any patient. Two patients had no change in drainage after remodelling and reimplantation of a megaureter, one was later diagnosed as having a neuropathic bladder and one child developed a UTI after ureteric reimplantation. The hospital stay was 3 days after pyeloplasty and 5 days after reimplantation. CONCLUSION: The design of the multipurpose Blue stent provides versatility; it can be used as a stent, and both an internal and external drain. Its use does not prolong the hospital stay. Insertion causes minimal trauma to the renal parenchyma, and removal is easy, pain-free and requires no anaesthesia. The complication rates in the present series compare favourably with other reported series. PMID- 9393308 TI - Evaluation of the utility of video-urodynamics in children with urinary tract infection and voiding dysfunction. AB - OBJECTIVE: To assess the use of video-urodynamic studies (VUDS) in children with urinary tract infection (UTI) and symptoms of voiding dysfunction (frequency, urgency, incontinence), to ascertain whether VUDS significantly assists in diagnosis and deciding treatment. PATIENTS AND METHODS: Over a 16-month period, all children seen at our centre with a UTI in conjunction with previous symptoms suggestive of voiding dysfunction underwent free and pressure-flow VUDS. Forty two children underwent VUDS and 38 (mean age 9 years, range 4-16, 15 male, 23 female) had sufficient information to be included in the study. RESULTS: All children had a prior history of voiding dysfunction (mean 55 months). Only five patients were found to have reflux and three of these had associated detrusor instability. In addition, 24 of 33 patients who did not have reflux had abnormalities on urodynamic study, the most common problem being detrusor instability in 17 of 24 patients. Other abnormalities included sphincter dyssynergia (five patients), poor bladder compliance (two) and hypersensitivity on bladder filling (three). CONCLUSION: VUDS can provide information about the aetiology of UTI and voiding dysfunction in children that cannot be obtained from any other source. The results of VUDS can be used to select specific treatments, to avoid inappropriate therapy and to identify children who may benefit from follow-up studies despite normal findings on voiding cystourethrography. From these results, we believe that VUDS should be considered for children with UTI and voiding dysfunction. PMID- 9393309 TI - A possible explanation of wet and dry nights in enuretic children. AB - OBJECTIVE: To clarify the relationship between nocturnal urine production and the occurrence of both wet and dry nights in patients with nocturnal enuresis and to estimate the effect on nocturnal urine production of treatment with the antidiuretic hormone desmopressin in a group of enuretics with none or only a partial reduction in the number of wet nights in response to desmopressin treatment. PATIENTS AND METHODS: The nocturnal urine production of 60 children with monosymptomatic nocturnal enuresis was measured for 14 nights with no treatment (baseline) and for 14 nights with desmopressin treatment. Sixteen children having both wet and dry nights in the two periods were chosen for the subsequent analysis. RESULTS: There was significantly less nocturnal urine production during desmopressin treatment (202 mL/night) than during the baseline period (279 mL/night; P < 0.001) and a corresponding decrease in the number of wet nights, from 10 during baseline to five during desmopressin treatment. When expressed as mL/kg body weight per hour, the urine production during baseline was 0.89 on wet and 0.625 on dry nights (P < 0.001), and during desmopressin treatment was 0.716 and 0.535, respectively (P < 0.01). CONCLUSION: In this group of enuretics there was a clear reduction in the number of wet nights and in nocturnal urine production during desmopressin treatment, even though none became totally dry on desmopressin. There was a markedly higher nocturnal urine production on wet nights during both the baseline period and during desmopressin treatment. The higher urine production on wet nights could explain the enuretic episode, with urine production exceeding bladder capacity. PMID- 9393310 TI - Surgery for cavoatrial extension of renal malignant tumours using a veno-venous shunt. PMID- 9393311 TI - Autoaugmentation in the paediatric neurogenic bladder: report of a method. PMID- 9393312 TI - Ureteroscopic salvage of a uretero-vaginal fistula. PMID- 9393313 TI - Neonatal bladder rupture due to anterior urethral valves. PMID- 9393314 TI - Ectopic labium in a newborn: a successful surgical transposition treatment. PMID- 9393315 TI - Glomus tumour of the testicle. PMID- 9393316 TI - Urodynamic evaluation of profound microcephaly in children. PMID- 9393317 TI - Caval leiomyosarcoma. PMID- 9393318 TI - Massive splenomegaly complicating left percutaneous renal surgery. PMID- 9393319 TI - Cardiac metastases from a transitional cell carcinoma: an unusual clinical manifestation. PMID- 9393320 TI - Extragonadal germ cell tumour involving the urinary bladder. PMID- 9393321 TI - Uretero-uterine fistula as a complication of caesarean section: successful ureteroscopic management. PMID- 9393322 TI - Comparison of real-time ultrasonography and magnetic resonance imaging in the assessment of urinary bladder volume. PMID- 9393323 TI - Fantasies and facts of testes. PMID- 9393324 TI - Congenital fistula of the penile urethra. PMID- 9393325 TI - British urological surgery practice: prostate cancer. PMID- 9393326 TI - A modified in vitro technique to evaluate human detrusor muscle. PMID- 9393327 TI - Urinary drainage systems to prevent intraluminal spread of bacteria. PMID- 9393328 TI - A standard protocol for the evaluation of laser treatment of the prostate. ICS 'BPH' Study Group. PMID- 9393329 TI - Assessing the odds. PMID- 9393330 TI - Still more questions on pertussis vaccines. PMID- 9393331 TI - New therapies for severe meningococcal disease but better outcomes? PMID- 9393332 TI - Balanced view of risks of oral contraceptives. PMID- 9393333 TI - Ultrasonography vs cystourethrography to exclude vesicoureteric reflux in babies. PMID- 9393334 TI - Risks and prevention of Dupuytren's contracture. PMID- 9393335 TI - Randomised controlled trial of two-component, three-component, and five-component acellular pertussis vaccines compared with whole-cell pertussis vaccine. Ad Hoc Group for the Study of Pertussis Vaccines. AB - BACKGROUND: Trials in Italy and Sweden showed high efficacy for three-component and five-component pertussis vaccines, and poor efficacy for a whole-cell vaccine licensed in the USA and a two-component vaccine. We compared the efficacy of three acellular vaccines with a UK whole-cell vaccine. METHODS: We enrolled 82,892 babies aged 2-3 months. Babies were vaccinated at age 3 months, 5 months, and 12 months, or age 2 months, 4 months, and 6 months. They were randomly assigned a two-component acellular diphtheria-tetanus-pertussis (DTP) vaccine (n = 20,697), a three-component acellular DTP vaccine (n = 20,728), a five-component acellular DTP vaccine (n = 20,747), or a UK whole-cell DTP vaccine (n = 20,720). We collected data for all reported cases of culture-confirmed pertussis during 3 years of follow-up. The treatment status of the two-component-vaccine group had to be made known midway through the trial for boosting because of poor efficacy. We included data for the two-component vaccine in the analysis of safety and immunogenicity, and data up its unmasking in secondary analyses of relative efficacy. Analyses were by intention to treat. FINDINGS: During follow-up from the third dose (mean 22 months), in the 3 months, 5 months, 12 months schedule, there were 15 cases of culture-confirmed pertussis with at least 21 days of paroxysmal cough in the whole-cell group, relative risk 1.00, compared with 13 in the five-component group (0.85 [95% CI 0.41-1.79]), and 21 in the three-component group (1.38 [0.71-2.69]). For culture-confirmed pertussis, with or without cough, there were 19 cases in the whole-cell group (1.00). 27 in the five-component group (1.40 [0.78-2.52]), and 49 in the three-component group (2.55 [1.50-4.33]). In the intention-to-treat analyses, from the first dose in the 3 months, 5 months, 12 months schedule the whole-cell vaccine was significantly more protective than the three-component vaccine against typical pertussis. Between the second and the third doses, culture-confirmed pertussis with any cough and with at least 21 days of paroxysmal cough was significantly more frequent in the two-component group than in the three-component group, and in the three-component group than in the five-component and the whole-cell groups, respectively. The serological response of the acellular vaccines in the 2 months, 4 months, 6 months schedule were similar to those previously reported. The whole-cell vaccine was highly immunogenic for fimbriae, pertactin, and filamentous haemagglutinin, but had a low antipertussis toxin response. Hypotonic hyporesponsiveness occurred significantly more frequently in the whole-cell group (p < 0.05) and was more frequent in the acellular groups than previously reported. High fever and seizures occurred more frequently after whole-cell vaccine than after any of the acellular vaccines (p < 0.001). INTERPRETATIONS: The efficacy of the UK whole cell vaccine and the five-component and three-component vaccines was similar against culture-confirmed pertussis with at least 21 days of paroxysmal cough. The lower efficacy of the three-component vaccine against mild disease suggests that fimbriae have a role in protection against infection. The efficacy of acellular vaccines depends on the number of components, and different whole-cell vaccines have variable efficacies. PMID- 9393336 TI - Lay diagnosis and health-care-seeking behaviour for chest pain in south Asians and Europeans. AB - BACKGROUND: South Asian people in the UK experience greater delays than Europeans in obtaining appropriate specialist management for heart disease, but the causes are not known. We investigated whether south Asians and Europeans interpret and act upon anginal symptoms differently. METHODS: We randomly selected 2000 people from general practitioners' (family physicians) lists in London, UK, to receive a questionnaire that included a short fictional case history of an individual with possible anginal pain and asked how respondents would react to experiencing it. A second questionnaire seeking information on medical history, attitudes to health, and demography was sent later. The main outcome measure was the proportion who said they would seek immediate care (hospital emergency department or general practitioner) for the pain described in the case scenario. FINDINGS: The rate of response to both questionnaires was 60.2% (903 of 1500 who received both), 553 responders were of European origin, 124 were Hindu, and 235 were Sikh. There were no differences between the ethnic groups in the proportion identifying the pain as cardiac, but south Asians would be more anxious about the pain than would Europeans. Of the men, 55 (23%) Europeans, 20 (38%) Hindus, and 52 (47%) Sikhs said they would seek immediate care (p < 0.0001 for heterogeneity); of women, 77 (24%), 25 (35%), and 58 (46%), respectively, would seek immediate care (p < 0.0001). After adjustment for confounding variables the odds ratio for seeking immediate care in Hindus compared with Europeans was 2.67 (95% CI 1.49-4.73) and that for Sikhs compared with Europeans was 3.18 (1.98-5.12). INTERPRETATION: Hindus and Sikhs reported a greater likelihood of seeking immediate care for anginal symptoms than Europeans; this finding indicates that barriers to cardiology services for south Asians are unrelated to difficulties in interpretations of symptoms or willingness to seek care. Improvement of awareness of heart disease may not decrease delays in obtaining care. Service-related explanations must be explored, such as general practitioners' difficulties in arriving at a diagnosis or differences in management because of ethnic origin. PMID- 9393337 TI - Origins of health inequalities in a national population sample. AB - BACKGROUND: Explanations for social inequalities in health are often explored but remain largely unresolved. To elucidate the origins of health inequalties, we investigated the extent to which adult-disease risk factors vary systematically according to social position over three decades of early life. METHODS: We used the 1958 birth cohort (all children born in England, Scotland, and Wales on March 3-9, 1958) with data up to age 33 years from parents, teachers, doctors, and cohort members (n = 11,407 for age 33 interview). FINDINGS: Social class of origin was associated with physical risk factors (birthweight, height, and adult body-mass index); economic circumstances, including household overcrowding, basic amenities, and low income; health behaviour of parents (breastfeeding and smoking) and of participants (smoking and diet); social and family functioning and structure, such as divorce or separation of participants or their parents, emotional adjustment in adolescence, social support in early adulthood; and educational achievement and working career, in particular no qualifications, unemployment, job strain, and insecurity. With few exceptions, there were strong significant trends of increasing risk from classes I and II to classes IV and V. Self-perceived health status and symptoms were worse in participants with lower class origins. INTERPRETATION: An individual's chance of encountering multiple adverse health risks throughout life is influenced powerfully by social position. Social trends in adult-disease risk factors do not emerge exclusively in mid life, but accumulate over decades. Investment in educational and emotional development is needed in all social groups to strengthen prevention strategies relating to health behaviour, work-place environment, and income inequality. PMID- 9393338 TI - Use of protein-C concentrate, heparin, and haemodiafiltration in meningococcus induced purpura fulminans. AB - BACKGROUND: Inflammatory and coagulation processes are both affected in meningococcaemia. Severe acquired protein-C deficiency in meningococcaemia is usually associated with substantial mortality: in survivors, skin grafts, amputation, and end-organ failure are not uncommon. Protein C is a natural anticoagulant and also has important anti-inflammatory activity. We assessed the effects of early replacement therapy with protein-C concentrate together with continuous veno-venous haemodiafiltration and conventional treatment in meningococcaemia. METHODS: 12 patients aged between 3 months and 27 years with meningococcaemia and severe acquired protein-C deficiency (mean 0.20 IU/mL) were studied. All patients had septic shock, widespread purpura, skin necrosis, and disseminated intravascular coagulopathy. After a test dose of protein-C concentrate, patients received a continuous infusion with the dose adjusted daily to keep the plasma concentration between 0.8 and 1.2 IU/mL. 11 patients were given unfractionated intravenous heparin (10-15 IU kg-1 h-1). Nine patients had haemodiafiltration and one had peritoneal dialysis. The Glasgow meningococcal septicaemia prognostic score and the paediatric risk of mortality score predicted a minimum mortality of 80% and 57%, respectively. FINDINGS: No patient died. No adverse reactions to the treatment were seen. Two patients had lower-limb amputations, one of whom had a thrombotic cerebrovascular accident; both patients had received the protein-C concentrate and heparin later than the rest of the group (60 h [16.97] vs 12 h [3.13]). One patient developed chronic renal failure despite receiving protein-C infusion 15 h after admission. INTERPRETATION: The acquired severe deficiency of protein C in meningococcaemia contributes to the pathogenesis of the thrombotic necrotic lesions in the skin and other organs and probably has an important role in the inflammatory response. Protein-C therapy is merely one approach to improve the host response in this syndrome. We suggest that a double-blind, randomised, controlled multicentre trial is needed to confirm our results. PMID- 9393339 TI - An infant with encephalitis. PMID- 9393340 TI - Early computed-tomography abnormalities in acute stroke. PMID- 9393341 TI - Benign symmetric lipomatosis associated with protease inhibitors. PMID- 9393342 TI - Spontaneous neutralising antibodies to interferon--alpha and interleukin-12 in thymoma-associated autoimmune disease. PMID- 9393343 TI - El Nino and diarrhoea and dehydration in Lima, Peru. PMID- 9393344 TI - Melatonin in feverfew and other medicinal plants. PMID- 9393345 TI - Leukotriene C4 synthase promoter polymorphism and risk of aspirin-induced asthma. PMID- 9393346 TI - Decreased circulating adrenomedullin in pre-eclampsia. PMID- 9393347 TI - Constrictive pericarditis after high-dose chemotherapy. PMID- 9393348 TI - Compatibility of insulin pens and cartridges. PMID- 9393349 TI - Bartonella-like erythrocyte inclusions in thrombotic thrombocytopenic purpura. PMID- 9393350 TI - World Bank oil-pipeline project designed to prevent HIV transmission. PMID- 9393351 TI - AIDS cost takes its toll in Canada. PMID- 9393352 TI - Jet-lag. PMID- 9393353 TI - What is required of an HIV vaccine? AB - Mounting evidence suggests that the early dissemination of HIV in human beings evokes an immune response that is responsible for containment of the infection during the long symptom-free period. Loss of this immune control coincides with a final escalation of the viraemia and the terminal failure of the immune system. Other studies imply that pre-emptive vaccination of monkeys with attenuated forms of simian immunodeficiency virus (SIV) produces a substantial degree of resistance to superinfection with fully virulent viruses. Here we consider how observations from natural and experimental systems might influence thought as to what is required to produce safe induced immunity against HIV. We concentrate on three questions: what is the nature of the immune response that contains the infection? How does this response fail? How could a vaccine enhance protective immunity so that it exceeds the efficacy of this natural response? PMID- 9393354 TI - Literature in the education of the doctor. PMID- 9393355 TI - Malarone-donation programme in Africa. PMID- 9393356 TI - Imperial Cancer Research Fund and the Lancet. PMID- 9393357 TI - Imperial Cancer Research Fund and the Lancet. PMID- 9393358 TI - Imperial Cancer Research Fund and the Lancet. PMID- 9393359 TI - Hormone replacement therapy and breast cancer. PMID- 9393360 TI - Hormone replacement therapy and breast cancer. PMID- 9393361 TI - Malarone-donation programme. PMID- 9393362 TI - Fetal nuchal translucency test for Down's syndrome. PMID- 9393363 TI - Fetal nuchal translucency test for Down's syndrome. PMID- 9393364 TI - Fetal nuchal translucency test for Down's syndrome. PMID- 9393365 TI - Fetal nuchal translucency test for Down's syndrome. PMID- 9393366 TI - Fetal nuchal translucency test for Down's syndrome. PMID- 9393367 TI - Treatment of hypertension in elderly patients. PMID- 9393368 TI - Treatment of hypertension in elderly patients. PMID- 9393369 TI - Treatment of hypertension in elderly patients. PMID- 9393370 TI - Obesity. PMID- 9393371 TI - Acts of violence against Rwandan refugees. PMID- 9393372 TI - Illegal drug use and HIV-1 infection in Columbia. PMID- 9393373 TI - Quality of drug advertisements in medical journals. PMID- 9393374 TI - How reliable is peer review? An examination of operating grant proposals simultaneously submitted to two similar peer review systems. AB - To determine level of agreement and correlation between two similar but separate peer review systems, proposals simultaneously submitted during the same funding year to two agencies using the same scoring system were identified and analyzed (n = 248). There was a direct linear relationship between the scores of the two agencies (r = 0.592, p < 0.001). Raw agreement within whole-digit ranges was moderate (53%) but a Cohen's kappa indicated that agreement beyond chance was only fair (kappa = 0.29, 95% CI = 0.198, 0.382). When proposals were arbitrarily categorized as being "clearly fundable" (on a 0-5 scale, score > or = 3.0) or "not clearly fundable" (score < 3.0), raw agreement was 73% and agreement beyond chance was moderate (kappa = 0.444, 95% CI = 0.382, 0.552). In cases where there was inter-agency disagreement on the fundability of the project, the difference in scores was greater than in those in which there was agreement. In a subsample of 128 pairs, variables describing the application and the applicant (i.e., principal investigator) were coded, but none explained inter-agency agreement on the "fundability" of proposals. PMID- 9393375 TI - Asymptomatic microscopic hematuria--is investigation necessary? AB - Microscopic hematuria is common in asymptomatic adults, but the benefit of screening the general population for blood in the urine has not been established. On the other hand, most studies of referred patients with putatively asymptomatic microscopic hematuria have reported a 2-11% prevalence of urothelial malignancies, leading to the recommendation that all patients with microscopic hematuria be thoroughly investigated. Urinalysis is inexpensive and highly acceptable to the general population, but is neither a sensitive, nor specific test, and has poor predictive value for urothelial malignancies, and nephrological diseases. Furthermore the benefits of early detection of such diseases has not been established. We conclude that screening urinalysis cannot be recommended. Studies are needed to determine which constellation of findings primary physicians use to select patients for referral to centers with urological and nephrological expertise. PMID- 9393376 TI - Problems in defining cutoff points of continuous prognostic factors: example of tumor thickness in primary cutaneous melanoma. AB - Continuous prognostic factors are often categorized by defining optimized cutoff points. One component of criticism of this approach is the problem of multiple testing that leads to an overestimation of the true prognostic impact of the variable. The present study focuses on another crucial point by investigating the dependence of optimized cutoff points on the observed distribution of the continuous variable. The continuous variable investigated was the vertical tumor thickness according to Breslow, which is known to be the most important prognostic factor in primary melanoma. Based on the data of 5093 patients, stratified random samples were drawn out of six artificially created distributions of tumor thickness. For each of these samples, Cox models were calculated to explore optimized cutoff points for tumor thickness together with other prognostic variables. The optimized cutoff points for tumour thickness varied considerably with the underlying distribution. Even in samples from the same distribution, the range of cutoff points was amazingly broad and, for some of the distributions, covered the whole region of possible values. The results of the present study demonstrate that optimized cutoff points are extremely data dependent and vary notably even if prerequisites are constant. Therefore, if the classification of a continuous prognostic factor is necessary, it should not be based on the results of one single study, but on consensus discussions including the findings of several investigations. PMID- 9393377 TI - Comparing dichotomous screening tests when individuals negative on both tests are not verified. AB - Two dichotomous screening tests are often compared by performing both tests in a sampled population, and submitting positive results on either test to verification by the reference standard. Unbiased estimates of the true positive and false positive rates of each test cannot be estimated directly. However, unbiased estimates of the relative true positive and relative false positive rates may be obtained. When one test has a higher true positive rate at the expense of a higher false positive rate, the trade-off is represented by the ratio of extra false positives detected to extra true positives detected. A 95% confidence interval for this ratio is derived. This ratio is prevalence dependent and only applies to the sampled population. For target populations of different prevalence, estimates of the ratio may be obtained if one of the following applies: (i) the test characteristics of one test are known; (ii) the relative prevalence is known; and (iii) certain assumptions are made. PMID- 9393378 TI - A logistic regression model when some events precede treatment: the effect of thrombolytic therapy for acute myocardial infarction on the risk of cardiac arrest. AB - When outcomes occur in clinical trials before treatment can be given, neither intent-to-treat nor according-to-protocol analyses give optimal estimates of the treatment effect. A better approach employs a time-dependent variable for treatment. Intent-to-treat analyses are conservative, biasing against treatment; according-to-protocol analyses bias in favor of treatment. We show how to measure the effect of a time-dependent variable in a logistic regression using person time intervals as units of measurement and describe appropriate methods for reporting model performance. The method is applied to develop a model to predict the probability that a patient with a myocardial infarction will have a sudden cardiac arrest within 48 hours of presentation to emergency medical services both when treated with thrombolysis and when not treated. We use a time-dependent treatment variable because many patients went into cardiac arrest while awaiting treatment. This technique has been programmed into an electrocardiograph for real time use in an emergency department. PMID- 9393379 TI - Predicting outcomes in HIV-infected veterans: I. Progression to AIDS. AB - This article and the following article (Parts I and II) report the development of two clinical staging systems for HIV-infected individuals. The objective of the research reported here (Part I) was to construct a clinical staging system to predict progression to AIDS. We analyzed data from VA Cooperative Study Number 298, a multicenter, double-blind, randomized trial that compared immediate versus deferred zidovudine therapy in 338 HIV-infected individuals who did not have AIDS at enrollment. Baseline variables were tested in univariate Cox regression for their relationship to progression to AIDS, and those that appeared predictive were examined in multivariable analysis. Based on these analyses, we constructed a new clinical staging system based on CD4+ cell count, age, hemoglobin, oral hairy leukoplakia or oral thrush, and fever. The stages of the system were significant predictors of progression to AIDS (p = 0.0001, log-rank test). In conclusion, simple, valid, clinical staging systems for HIV-infected patients can be constructed using information that is readily available in clinical practice settings. Such systems provide better prognostic distinction than CD4+ cell count alone by taking into account the known prognostic effects of other variables. PMID- 9393380 TI - Predicting outcomes in HIV-infected veterans: II. Survival after AIDS. AB - This article (Part II) and the preceding article (Part I) report the development of two clinical staging systems for HIV-infected individuals. The objective of the research reported here (Part II) was to construct a clinical staging system to predict survival in patients with AIDS. We analyzed data from VA Cooperative Study Number 298, a multicenter, double-blind, randomized trial that compared immediate versus deferred zidovudine therapy in HIV-infected individuals. Baseline variables obtained at the onset of AIDS in 204 individuals were tested in univariate Cox regression for their relationship to survival, and those that appeared predictive were examined in multivariable analysis. Based on these analyses, we constructed a new AIDS Clinical Staging System. The system is based on age, CD4+ cell count, type of first AIDS-defining condition, and functional status. The stages of the system were significant predictors of survival (p = 0.0001, log-rank test). In conclusion, valid, simple clinical staging systems for patients with AIDS can be developed based on a few variables that are readily available in clinical settings. PMID- 9393381 TI - The effect of estrogen replacement therapy on cognitive function in women: a critical review of the literature. AB - OBJECTIVE: To conduct a review of the available clinical trials to determine whether sufficient evidence exists to support the conclusion that estrogen replacement therapy has a beneficial effect on cognitive performance in post menopausal women and in women with Alzheimer's disease. Studies were identified through a MEDLINE search of all English-language publications between 1970 and 1996 in which the words estrogen and cognition or estrogen and memory appeared. DATA EXTRACTION: Data were extracted for each study, including features of subjects and eligibility criteria, duration of follow-up, and treatment regimen. Baseline characteristics were evaluated, including age; menopausal status; follicle-stimulating hormone, luteinizing hormone, and estradiol levels; mood; and measures of cognitive function. Psychological tests were evaluated for construct validity. RESULTS: Nineteen studies were reviewed, including 10 randomized trials of estrogen replacement therapy versus placebo. Extreme heterogeneity among subjects and variability in the use of cognitive measures across the studies precluded performing a quantitative summary. Of the 10 randomized trials, eight claimed therapeutic benefits for estrogen therapy, three of which reported significant improvements in memory and two of which showed improvements in attention. These studies did not control for potential confounds such as depression and vasomotor symptoms. Of the nine observational studies, five found a significant association between estrogen use and cognitive function. CONCLUSION: Although several observational studies provide encouraging evidence for the beneficial effect of estrogen on cognitive function, there is currently inadequate evidence available from randomized, controlled trials to support the conclusion that estrogen replacement therapy improves cognitive function in post menopausal women or women with Alzheimer's dementia. PMID- 9393382 TI - Use of physician services among family caregivers of elderly persons with dementia. AB - It is well known that there is an excess of physical and psychological health problems among family caregivers of elderly persons with Alzheimer's disease and other dementias. The objective of this study was to determine whether the higher level of morbidity translates into a higher level of medical care utilization. Data from a previously completed longitudinal study of caregivers for elderly persons with dementia were merged with data on physician visits obtained from the computerized records of the Quebec Health Insurance Board. Utilization of physician care (adjusted for age, sex, number of chronic diseases, and depression) was no higher for family caregivers of elderly patients with Alzheimer's disease and other dementias than for comparable family members of older persons without dementia. The annual cost of physician care was almost identical among caregivers and noncaregivers. However, the pattern of utilization for the two groups was somewhat different: there was a significantly higher frequency of physician utilization among caregivers for services billed by psychiatrists and internal medicine specialists. In multivariate analysis, physician utilization was significantly associated with having more than one chronic condition and with increased age. Future studies should focus on determining whether caregivers neglect their own health care needs as a result of the exigencies of the caregiving role. PMID- 9393384 TI - Interpretation and bias in case-crossover studies. AB - The case-crossover design is an innovative epidemiologic technique with distinct strengths and limitations. We review the fundamental logic of this self-matching non-randomized design and direct attention to 15 concerns related to the available data, unavailable data, analytic technique, quantitative statistics, and etiologic model. Implications for each concern are discussed in the context of a recent report on whether cellular telephone calls are associated with an increased risk of a motor vehicle collision. We suggest that an understanding of the case-crossover design may help investigators explore selected questions in behavioral medical research. PMID- 9393383 TI - Statistical evaluation of the role of Helicobacter pylori in stress gastritis: applications of splines and bootstrapping to the logistic model. AB - Stress gastritis is a serious problem in the intensive care unit population. The recent discovery of the causal nature of Helicobacter pylori (H. pylori) in the development of gastric ulcers led us to examine its relationship with stress gastritis. We investigated this relationship in 874 veterans admitted to intensive care units who were tested for the presence of H. pylori and followed for 6 weeks for the development of stress gastritis. We fit spline models to assess functional relationships and used the logistic model to determine the association between H. pylori and stress gastritis. The predictive ability of the model was assessed with receiver operating characteristic (ROC) curve analysis and validated with the bootstrapping technique. Increased anti-H. pylori immunoglobulin A concentrations were found to be an important predictor of stress gastritis independent of other known risk factors. PMID- 9393385 TI - Ethnicity and conditional breast cancer survival in Hawaii. AB - The conditional survival experience of 4502 female breast cancer cases diagnosed in Hawaii between 1960 and 1983 was studied. The calculation of conditional survival is based on patients who have already survived for a specified time period. Cox proportional hazards models were used to examine the effect of ethnicity, stage at diagnosis, and menopausal, marital, and socioeconomic statuses on conditional survival prognosis. Native Hawaiian and Filipino women showed a significantly higher risk of dying from breast cancer than any other ethnic group, followed by Chinese and Caucasian women, while Japanese women had the lowest risk. The ethnic pattern of breast cancer survival changed and ethnic survival differences decreased from 17 to 4% as time progressed. Conditional survival varied greatly with stage at first diagnosis. For localized, regional, and distant disease, the conditional survival rate increased from 92, 71, and 24% after diagnosis to 95, 85, and 65% 5 years after diagnosis, respectively, indicating that stage at diagnosis becomes less important in determining survival as time progresses. Being married and of a high socioeconomic status increased survival probability. These results suggest that considering time effect, the conditional survival rate is an informative tool to assess clinical outcome of breast cancer among ethnic groups over a long follow-up period. PMID- 9393386 TI - Antibiotic noncompliance and waste in upper respiratory infections and acute diarrhea. AB - A prospective cohort study was conducted to analyze factors associated with antibiotic noncompliance and waste among patients suffering acute respiratory infection (ARI) and acute diarrhea (AD). The study took place in four primary health care clinics in Mexico City, two belonging to the Ministry of Health (MoH) and two to the Mexican Social Security Institute (IMSS). Two hundred twenty-two patients with ARI and 155 with AD were included. Data about study variables and the assessment of compliance were obtained through patient interviews and direct observation. Factors associated with noncompliance were assessed through a multiple logistic regression procedure. Noncompliance was 60% for ARI and 55.5% for AD in both health care systems. Prescription of an antibiotic was justified only in 13.5% of cases. Associated factors were: increased duration of illness (OR 2.95; 95% CI, 1.17-7.41); complexity of the treatment: 3 or more doses per day (OR 2.47; 95% CI, 1.56-3.92), and treatment for more than 7 days (OR 1.94; 95% CI, 1.16-3.26); younger age of patient (OR 1.89; 95% CI, 1.18-3.02); and an inadequate physician-patient relationship (OR 1.87; 95% CI, 1.16-3.02). Antibiotic waste was higher in IMSS (ARI 39.3%, AD 32.6%), than in the MoH (ARI 21.2%, AD 16.4%). Educational strategies to modify physician prescribing practices and strengthen physician-patient relationships might improve compliance and decrease drug waste. PMID- 9393387 TI - Quantification and the quest for medical certainty. PMID- 9393388 TI - Neural networks and logistic regression: analysis of a case-control study on myocardial infarction. PMID- 9393389 TI - Obsessive-compulsive disorder: the hidden epidemic. PMID- 9393390 TI - Epidemiology of obsessive-compulsive disorder: a world view. AB - The worldwide prevalence of obsessive-compulsive disorder (OCD) is approximately 2% of the general population. Symptoms of OCD include fear of contamination by dirt or germs; constant checking; repetitive, intrusive thoughts of a somatic, aggressive, or sexual nature; extreme slowness; and an inordinate concern with orderliness and symmetry. Differential diagnosis is sometimes complicated by the overlap between OCD and obsessive-compulsive personality disorder (OCPD). The most common complication of OCD is depression. However, while both serotonergic and nonserotonergic antidepressants are effective in treating patients with depression, only serotonergic medications are effective in treating OCD patients. Because OCD patients often attempt to conceal their symptoms, it is incumbent on clinicians to screen for OCD in every mental status examination, since appropriate treatment can often result in improved quality of life. PMID- 9393391 TI - Update on pharmacologic management of OCD: agents and augmentation. AB - A preponderance of patients with obsessive-compulsive disorder (OCD) experience little or no improvement in their symptoms when treated with serotonin reuptake inhibitors (SRIs). It is hypothesized that SRI-refractory patients may have altered serotonin neurotransmission different from patients responsive to SRIs, or that they may have abnormalities in their dopamine function. When drugs affecting serotonin function (e.g., tryptophan, fenfluramine, lithium, buspirone) are added to SRI therapy in SRI-refractory patients, results are mixed and not consistently encouraging. However, when drugs affecting dopamine function (e.g., pimozide, haloperidol, risperidone) are added to SRI therapy in SRI-refractory OCD patients, individuals with either a personal history or family history of tics experience a reduction in their symptoms. PMID- 9393392 TI - Pharmacologic treatment of obsessive-compulsive disorder: comparative studies. AB - The predominant hypothesis about obsessive-compulsive disorder (OCD) pathophysiology implicates abnormal serotonergic function regulation. Pharmacologic agents with potent serotonin reuptake-inhibiting properties have demonstrated effectiveness in treating OCD. In short-term clinical trials compared by meta-analysis, clomipramine and serotonin selective reuptake inhibitors (SSRIs) were found superior to placebo in improving symptoms of OCD. In one-to-one comparative studies, clomipramine has been found as efficacious as fluoxetine and fluvoxamine, and in a comparative trial of clomipramine with sertraline, there was a statistically superior response to sertraline after 16 weeks of treatment; moreover, discontinuation rate in patients taking clomipramine was more than twice that in patients taking sertraline (26% vs. 11%). In contrast to patients receiving clomipramine who showed poor tolerance in long-term use, patients maintained on fluoxetine for 24 weeks after an acute phase well tolerated the medication. In another study, patients responding to 12 weeks of sertraline treatment also showed improved tolerance during an additional 40-week period, with 75% completing the continuation phase. With long-term or even lifelong treatment appearing necessary for people with OCD, those agents that result in better tolerance will prove preferable. PMID- 9393394 TI - Implantology 1997--do we have all the answers? PMID- 9393393 TI - An interactive computer-administered self-assessment and self-help program for behavior therapy. AB - Behavior therapy for obsessive-compulsive disorder (OCD) is extremely effective and usually conveys lasting benefits. Unfortunately, behavior therapy is not widely available and is usually quite costly in the few settings where it can be obtained. The essential features of effective behavior therapy are identifying triggers of obsessions, rituals, and discomfort; designing personalized exposure and ritual prevention (E & RP) goals; and encouraging and monitoring E & RP sessions of sufficient frequency and duration for habituation to occur. A computer program (BT STEPS) was designed to assist OCD sufferers in carrying out self-assessment and self-help behavior therapy. The program has nine clinical steps, 12 computer-controlled interactive voice response (IVR) telephone calls (some used repeatedly), and more than 1000 digitized voice files that depend on the progress that patients report during calls. BT STEPS has been studied in two trials at three sites with a total of 65 patients. In both trials, patients who experienced behavior therapy had substantial reductions in OCD severity as assessed by the Yale-Brown Obsessive Compulsive Scale. Approximately two thirds of those who participated in two or more E & RP sessions were much or very much improved. Patients liked using the program, and 71% thought their lives improved as a result. BT STEPS makes behavior therapy for OCD patients potentially available to anyone with a touch-tone telephone. It is intended for use under the supervision of a clinician and can be used in conjunction with pharmacotherapy. PMID- 9393395 TI - Breathing patterns and levels of consciousness in children during administration of nitrous oxide after oral midazolam premedication. AB - PURPOSE: The combination of midazolam and nitrous oxide is commonly used to achieve sedation and analgesia during pediatric oral procedures, yet there are few, if any, data that illustrate the ventilatory effects of N2O in children, especially when used in combination with additional central nervous system (CNS) depressants. It was hypothesized that the addition of N2O inhalation to oral midazolam premedication would enhance the sedative effects of the midazolam and add analgesia without causing significant respiratory depression. The purpose of this study was to test this hypothesis. MATERIALS AND METHODS: Thirty-four healthy children about to undergo restorative dental treatment under general anesthesia were premedicated with oral midazolam, 0.7 mg/kg, and were then exposed to 40% N2O for 15 minutes after a 5-minute control period. The effect of adding N2O on SpO2, respiratory rate, PETCO2, VT, and VT/TI was examined and the levels of consciousness (conscious vs deep sedation) before and during N2O inhalation were determined. RESULTS: During the course of the study, no child developed hypoxemia (SpO2 < 92%) nor clinically significant upper airway obstruction. Four children who did not develop hypoventilation (defined as PETCO2 > 45 mm Hg) during the control period did so after initiation of N2O. Overall, there were no significant differences in SpO2, PETCO2, VT, or VT/TI between the control and study periods. However, respiratory rates were significantly higher in the first 10 minutes of N2O inhalation when compared with the control period. Before starting N2O administration, 14 children were not clinically sedated, 19 children met the criteria for conscious sedation, and one child met the criteria for deep sedation. At the end of 15 minutes of N2O inhalation, 12 children were not clinically sedated, 17 children met the definition of conscious sedation, three were deeply sedated, and one child had no response to IV insertion, implying a state of general anesthesia. There were no differences in sedation scores between the control and study periods (P = .6). Overall, seven children had an increase in their sedation score while breathing N2O, four had a decrease in their sedation score, and 22 had no change. CONCLUSIONS: The addition of 40% N2O to oral midazolam, 0.7 mg/kg, did not result in clinically meaningful respiratory depression nor upper airway obstruction, but did, in some children, cause an increase in the level of sedation beyond simple conscious sedation. PMID- 9393396 TI - Recommendations for management of trigeminal nerve defects based on a critical appraisal of the literature. AB - PURPOSE: Management of trigeminal nerve injuries continues to challenge oral and maxillofacial surgeons. The purpose of this review article is to apply the principles of evidence-based medicine (E-BM) to determine the optimal operative technique for managing defects involving the inferior alveolar (IAN) or lingual nerves when direct (ie, primary) repair is not feasible. METHODS: To address the research purpose, the four steps of the E-BM critical appraisal process were applied: 1) identify the clinical problem, 2) efficiently search the literature, 3) select relevant articles and apply rules of evidence, and 4) apply the findings to patient care. Parameters for the literature search included using Medline to identify English language articles, publication dates from 1986 through 1996, and studies involving human subjects. RESULTS: The studies reviewed showed that the clinical literature on operative management of trigeminal nerve injuries is sparse, preoperative and postoperative neurosensory examinations are poorly documented, and the data are derived completely from reports using case series methods. Given these limitations, the available literature suggests that 1) tension-free, primary (direct) suture repair of an injured nerve, if possible, provides optimal results; 2) if direct repair is not possible, autogenous nerve grafts should be used for acute injuries, for example, immediate nerve repair after tumor resection or at the time of acute repair after traumatic injury; and 3) if direct repair is not possible, autogenous nerve grafts or hollow conduits (entubulization) to bridge the defect are equally successful for delayed reconstruction of gaps of 3 cm or smaller. CONCLUSIONS: Based on the weakness of the current literature, recommendations for future research include 1) better standardization and documentation of sensory deficits resulting from nerve injuries and their recovery, 2) using multicenter studies to accumulate large samples of patients rapidly, 3) using case series or prospective cohort study designs to assess the value of operative management of nerve injuries, and 4) progressing to randomized clinical trials to ascertain the optimal operative management of nerve injuries. PMID- 9393397 TI - Diagnosis and management of blunt carotid artery injury in oral and maxillofacial surgery. AB - PURPOSE: Traumatic occlusion of the internal carotid artery (ICA) is a rare complication of maxillofacial trauma or surgery. This investigation evaluated patient demographics, diagnostic methods, and effective therapeutic modalities associated with blunt carotid injury (BCI). PATIENTS AND METHODS: This was a retrospective analysis of patient records with an ICD-9-CM diagnosis of carotid injury conducted at MetroHealth Medical Center during the 24-month period between August 1993 and July 1995. Carotid injuries attributable to penetrating trauma were excluded. Age, gender, cause of injury, Glasgow Coma Scale score, Injury Severity Score, type and location of injury, concomitant injury, diagnostic methods, treatment modalities, and outcome were identified, recorded, and analyzed. RESULTS: During the 24-month period, 12 patients (seven males and five females) suffered BCI. These patients were divided into two groups based on cause of the problem. In group I, there were 3,214 blunt trauma patients admitted during the 2-year study, of which 10 patients had BCI, representing 0.31% of blunt trauma patients, and 1.2% of patients with head injuries. Seven patients presented with hemiplegia, two with cranial nerve palsy, and one with perceptual neglect. Ninety percent of the patients had associated injuries. Two patients had surgical intervention, three received anticoagulation, and five had only supportive care. Four of the 10 patients died, four had moderate neurologic deficits, and two survived with only minor neurologic deficits. In group II, two patients developed BCI after surgery. A 52-year-old woman had a carotid injury after right total temporomandibular joint replacement, and a 48-year-old man who underwent surgical removal of a third molar became hemiplegic postoperatively. The first patient recovered after anticoagulation, whereas the second patient, who received only supportive care, has severe neurologic deficits. CONCLUSIONS: BCI is an uncommon entity. It is usually recognized when a patient develops an unexplained neurologic deficit, most often hemiplegia, subsequent to trauma or surgery of the head, face, or neck. In the early stages, the diagnosis can be missed by carotid ultrasound or computed tomography. The injury is unrelated to Glasgow Coma Scale score. Symptoms may not develop for days after injury in 50% of patients. Anticoagulation appears to be the most beneficial therapeutic modality. PMID- 9393398 TI - Reconstruction of the severely resorbed maxilla with a combination of sinus augmentation, onlay bone grafting, and implants. AB - PURPOSE: A new method of reconstruction of the atrophic maxilla by combining a bilateral sinus floor elevation and cancellous bone graft with buccal and labial onlay graft using L-shaped corticocancellous blocks from the posterior iliac crest is presented. PATIENTS AND METHODS: Seventeen patients were treated with this procedure. One hundred one IMZ implants were placed in 14 patients, and 22 Branemark implants were placed in three patients. Patients were observed for 6 months after prosthetic rehabilitation. RESULTS: All patients were fully rehabilitated with fixed bridges except one, who preferred an overdenture. Only two implants were lost at the time of the abutment connection. Some bone resorption was seen around six implants. The success rate with this procedure was 92.7% 6 months after prosthetic rehabilitation if implants with bone resorption were considered failures. CONCLUSIONS: These preliminary results indicate that this surgical procedure is suitable for reconstruction of most atrophic maxillas. PMID- 9393399 TI - Treatment of mandibular fractures with rigid osteosynthesis: using the AO system. AB - PURPOSE: This study evaluated the results achieved in the surgical treatment of all mandibular fractures at two university centers using the 2.7 AO mandibular system. PATIENTS AND METHODS: A total of 227 patients presenting with 180 single fractures and 47 with double fractures (274 osteosyntheses) were included in this prospective study. RESULTS: During a mean follow-up of 27.5 months (minimum, 12 months), an overall complication rate of 7% was observed. No infection justified early removal of the osteosynthesis material. CONCLUSIONS: The systematic use of the technique recommended by AO for treating mandibular fractures, performed by thoroughly experienced operators on a compliant population, results in a low rate of complications and an early return to normal function. PMID- 9393400 TI - Use of activator appliances in pediatric patients treated with costochondral grafts for temporomandibular joint ankylosis: analysis of 13 cases. AB - PURPOSE: The long-term outcomes and clinical results of costochondral transplants used for the treatment of condylar ankylosis of the mandible in children with and without application of postoperative activator appliances are evaluated and compared. MATERIALS AND METHODS: A nonrandomized, retrospective clinical study of 13 cases of condylar ankylosis (16 joints) of the mandible surgically treated during a 9-year period from 1988 to 1997 was performed. All 13 patients were treated by condylectomy and immediate costochondral rib grafts. Nine of these patients underwent long-term postoperative therapy using removable activator appliances. Four patients did not undergo activator therapy postoperatively. Casts, radiographs, photographs, computed tomography (CT) scans, magnetic resonance imaging (MRI) and 99Tc bone scans were used postsurgically to evaluate graft take, condylar growth and function, occlusion, and facial and condylar symmetry. RESULTS: The postoperative and long-term clinical results in both groups showed costochondral growth center transplants to be effective in restoring mandibular growth of the affected side. However, symmetry, arch coordination, correction of occlusal canting, mandibular deviation, facial growth, and prevention of reankylosis were obtained and better controlled only in those cases that underwent long-term orthodontic activator therapy postoperatively and were followed closely. CONCLUSIONS: Children with long standing condylar ankylosis of the mandible and its resultant facial asymmetry and occlusal canting (secondary to a nonfunctional joint and maxillary compensation) treated with condylectomy and immediate costochondral rib graft reconstruction of the affected joint were treated more favorably when activators were used postsurgically. The patients that failed to comply with or continue activator therapy postsurgically developed complications relating to mandibular deviation, occlusal dysharmony, asymmetry and, in one case, reankylosis of the temporomandibular joint (TMJ). PMID- 9393401 TI - The free fibula bone graft for salvaging failed mandibular reconstructions. AB - PURPOSE: The purpose of this study was to determine the efficacy of vascularized free fibula bone grafts for mandibular salvage reconstruction. PATIENTS AND METHODS: Seven patients had fibula grafts after failed attempts at mandibular reconstruction. The prior attempts involved 20 operative procedures. Four of the seven patients (57%) had a history of radiation to the affected mandible. Bony defects averaged 10.2 cm (range, 4.5 to 24 cm), and the associated soft tissue defects averaged 6 x 12 cm. Average follow-up was 16 months. Cosmetic (facial symmetry) and functional (speech quality, oral continence, deglutition, donor site morbidity, dental rehabilitation) results were evaluated by questionnaire and clinical examinations. RESULTS: Soft tissue coverage and mandibular restoration were successful in all patients, and flap survival was 100%. Five of the seven patients (70%) achieved good or excellent functional results, and five of seven (70%) achieved good or excellent esthetic results. Complications were minimal, and the average hospital stay was 14 days. CONCLUSIONS: When the initial attempt at mandibular reconstruction is unsuccessful, mandibular function and esthetics can be salvaged with reliable vascularized soft tissue and bone flaps. As long as appropriate flap options are considered and the patient is medically stable, successful mandibular reconstruction should be achievable despite the number or cause of prior failed attempts. PMID- 9393402 TI - Experimental study of epithelialization of the muscle-only flap in the oral cavity. AB - PURPOSE: The purpose of this study was to observe the epithelialization process of the muscle-only flap used for reconstruction of the oral mucosal defects. MATERIALS AND METHODS: Forty-three male adult Japanese rabbits were used. A superiorly based cleidomastoid muscle flap was designed after vascular assessment. The flap was transferred into the oral cavity to cover a mucoperiosteal defect made in the mandibular alveolus. Epithelialization of the flap was histologically evaluated at designated intervals. RESULTS: The flaps survived without ischemic necrosis. By 8 days postoperation, the flap was infiltrated by acute inflammatory cells and being replaced by granulation tissue originating from the adjacent tissues. The oral epithelial cells advanced onto this granulating muscle flap, with eventual coverage by 21 days. The granulation tissue matured to fibrous tissue with significant contraction by 2 months. At 6 months postoperation, abnormally hyperkeratinized epithelium was seen on the flap. This differed from the surrounding parakeratinized oral epithelium. CONCLUSIONS: The muscle-only flap in the oral cavity epithelializes after the granulation process. PMID- 9393403 TI - Skeletal and dental movements after anterior maxillary advancement using implant supported distraction osteogenesis in dogs. AB - PURPOSE: This study evaluated skeletal and dental relapse in the dog after anterior maxillary advancement using an implant-anchored distraction osteogenesis device. MATERIALS AND METHODS: After the placement of implants into the palate, four dogs had a skeletally anchored distraction device fabricated and an anterior maxillary osteotomy performed. The distraction device was activated 0.5 mm two times each day to advance the anterior segment 10 mm in 10 days. The dental and skeletal changes were measured and compared for 26 weeks after distraction was completed. RESULTS: Tooth and radiographic measurements indicated that on the 10th day of distraction the average tooth advancement was 6.83 mm +/- 1.75 mm SD, and the average skeletal advancement, based on radiographs, was 8.67 mm +/- 1.25 mm SD. After 10 weeks, the average tooth advancement was 4.0 +/- 1.73 mm, and the average skeletal advancement was 8.67 +/- 3.59 mm. After 26 weeks, the average tooth advancement was 3.65 +/- 1.69 mm, and the average skeletal advancement was 8.0 +/- 1.98 mm. Bone healing occurred in all animals. The skeletal advancement 26 weeks after distraction was 85% of the initial advancement. The teeth adjacent to the distraction site initially moved 68% of the advancement, but the distance between the teeth subsequently decreased, with a resultant 36% advancement compared with the initial advancement. CONCLUSION: This study showed that skeletal fixation will result in bone movement greater than dental movement, indicating the need for skeletal anchorage to advance the maxilla. PMID- 9393404 TI - Repair of craniofacial defects with hydroxyapatite cement. AB - PURPOSE: The objective of this study was to evaluate the course of healing of craniofacial bone defects when filled with hydroxyapatite cement and to determine whether adding various percentages by weight of demineralized bone powder to the cement will result in enhanced bone formation. MATERIALS AND METHODS: The model for the study was the canine calvarium. The implants were placed into cranial defects and harvested at 3 or 6 months for qualitative evaluation by light microscopy, microradiography, and quantitative histomorphometry. RESULTS: The implantation of hydroxyapatite cement resulted in characteristic replacement of the material with new bone ingrowth. The addition of demineralized bone powder to the hydroxyapatite cement appeared to improve the handling characteristics of the cement; however, improvement in the replacement of the material by bone was not observed. The implantation of only allogeneic demineralized bone showed limited new bone formation within the defect site. CONCLUSIONS: Hydroxyapatite cement formed an effective osseoconductive scaffold for bone replacement. The addition of demineralized bone powder to the cement to serve as a carrier of osseoinductive factors did not result in additional bone being formed. PMID- 9393405 TI - Alcohol withdrawal syndrome: current management strategies for the surgery patient. AB - PURPOSE: As advances in the therapeutic management of alcohol withdrawal syndrome occur, oral and maxillofacial surgeons should be aware of the current treatment philosophies and modalities. This article provides a comprehensive review of alcohol withdrawal syndrome and presents some of the current management strategies that can be used for these patients, whether it be in the office or in the hospital. PMID- 9393406 TI - Mixed radiolucent/radiopaque lesion of the mandible. PMID- 9393408 TI - Melanoma of the tongue: case report. PMID- 9393407 TI - Adenoid cystic carcinoma originated from an anterior lingual minor salivary gland: immunohistochemical and ultrastructural studies and review of the literature. PMID- 9393409 TI - Bone-forming pleomorphic adenoma of the upper lip: report of a case. PMID- 9393410 TI - Orbital root blow-in fracture: report of a case. PMID- 9393411 TI - Disseminated intravascular coagulation associated with parotitis: a case report and review. PMID- 9393412 TI - Numb chin syndrome leading to a diagnosis of acute lymphoblastic leukemia: report of a case. PMID- 9393413 TI - Pseudoaneurysm of the internal maxillary artery and Frey's syndrome after blunt facial trauma. PMID- 9393415 TI - Osteoplasty and advancement genioplasty for widening of the chin. PMID- 9393414 TI - Crestal window technique for anterior iliac crest graft procurement. PMID- 9393416 TI - Ligation of the descending palatine artery: pro and con. PMID- 9393417 TI - Going back to our roots. PMID- 9393418 TI - The debate about debris. PMID- 9393419 TI - Clinical factors of importance to successful implant therapy. Preface. PMID- 9393420 TI - Clinical factors of importance to successful implant therapy. Introduction. PMID- 9393421 TI - Positive effect of surgical experience with implants on second-stage implant survival. AB - This Dental Implant Clinical Research Group study defined a learning curve for dental implant placement. Implants placed by inexperienced surgeons (< 50 implants) failed twice as often as those placed by experienced surgeons (> or = 50 implants). Implants placed during the first 6, 8, 10, 12, and 16 cases were compared with all others. The greatest difference was seen between the first nine cases and all others (P = .001), with later cases failing significantly less often. Inexperienced surgeons had more failures in the first nine cases (5.9%) than more experienced surgeons (2.4%). Surgeons with little or no previous experience must expect a definite learning curve. Previous experience may transfer and result in a shallower learning curve for subsequent systems. PMID- 9393422 TI - The influence of preoperative antibiotics on success of endosseous implants up to and including stage II surgery: a study of 2,641 implants. AB - According to the American College of Surgeons, complex oral surgical procedures, including the transoral placement of endosseous implants, are of the type that may require prophylactic antibiotics. However, the routine use of prophylactic antibiotics in the field of dental implantology continues to be controversial, and their utilization varies widely. No data from a randomized prospective clinical study of the prophylactic use of antibiotics in implant surgery have been previously published. As part of the comprehensive Dental Implant Clinical Research Group clinical implant study, the preoperative or postoperative use of antibiotics, the type used, and the duration of coverage was left to the discretion of the surgeon. These data were recorded and correlated with failure of osseointegration during healing (stage I) and at stage II surgery (uncovering). The results showed that significantly fewer failures occurred when preoperative antibiotics were used. PMID- 9393423 TI - The influence of 0.12% chlorhexidine digluconate rinses on the incidence of infectious complications and implant success. AB - The effect of perioperative chlorhexidine on the frequency of infectious complications through stage II was examined. Chlorhexidine was used perioperatively in 54.6% of patients (52.5% of implants) in a Dental Implant Clinical Research Group study with a database of 2,641 implants (595 patients). With chlorhexidine, there was a significant reduction in the number of infectious complications (4.1% vs 8.7%). Two percent of implants failed in the absence of an infectious complication, whereas 12% with infectious complications failed. This sixfold difference is highly significant. Chlorhexidine may reduce microbial complications when used in the immediate perioperative period. PMID- 9393424 TI - The influence of type of incision on the success rate of implant integration at stage II uncovering surgery. AB - In 1991, the Dental Implant Clinical Research Group comprising 30 Department of Veterans Affairs medical centers and two dental schools initiated a long-term clinical study to investigate the clinical performance of implants within the Spectra-System (Core-Vent Corporation, Las Vegas, NV). This article focuses on a portion of the study database related to incision type, implant success rates, and response of crestal bone up to the time of surgical uncovering. The crestal incision was used for 1,705 implants (381 patients) and the remote incision for 593 implants (141 patients). No statistically significant difference (P = .092 chi-square statistic) was found in implant integration or the response of crestal bone. PMID- 9393425 TI - Distribution of bone quality in patients receiving endosseous dental implants. AB - Knowledge of the distribution of bone quality in the various jaw regions assists the clinician in dental implant treatment planning. Bone quality was assessed with radiographs and tactile sensation for 2,839 implants at the time of placement into four anatomic regions of the jaw. The Lekholm-Zarb classification system was used. Overall, bone quality types 1 and 4 were found much less frequently than types 2 and 3. Although variations in density existed in each region, quality 2 bone dominated the mandible, and quality 3 bone was more prevalent in the maxilla. For both anterior and posterior jaw regions, types 2 and 3 bone predominated. The anterior mandible had the densest bone, followed by the posterior mandible, anterior maxilla, and posterior maxilla. PMID- 9393426 TI - Bone quality and implant design-related outcomes through stage II surgical uncovering of Spectra-System root form implants. AB - Failure rates at second-stage surgery were reported for the ongoing Dental Implant Clinical Research Group studies of the Spectra-System (Core-Vent Corporation, Las Vegas, NV) implants. As of May 1995, 69 implants failed out of 2,633 placed and uncovered. The overall failure rate was 2.6%, with 3.6% in bone quality 1 (BQ-1), 2.4% in BQ-2, 2.5% in BQ-3, and 3.1% in BQ-4. HA-coated cylinders had the lowest number of failures and titanium alloy baskets the highest. The basket design failed more often in the posterior jaw areas; 9 of 32 clinical centers (28%) accounted for 72% of these failures. PMID- 9393428 TI - Radiographic assessment of peri-implant vertical bone loss: DICRG Interim Report No. 9. AB - Vertical bone loss is being assessed radiographically as part of the Dental Implant Clinical Research Group studies through direct measurements on study radiographs taken longitudinally at surgery and recall appointments. Preliminary results and trends for the period between implant placement and 6 months after implant uncovering show more bone loss in implants that are 1) not coated with hydroxyapatite; 2) placed in the maxilla; 3) placed in anterior regions of the jaws; 4) in completely edentulous cases; and 5) placed in bone scored as having lower quality. Confounding relationships between predictor variables will require controlled statistical analyses when data collection is completed. PMID- 9393427 TI - The influence of bone quality on Periotest values of endosseous dental implants at stage II surgery. AB - Periotest values (Periotest, Siemens AG, Bensheim, Germany) were recorded as a baseline variable at surgical uncovering in the ongoing multicenter, prospective clinical studies of the Dental Implant Clinical Research Group, which uses implants from the Spectra-System (Core-Vent Corporation, Las Vegas, NV). For 2,212 osseointegrated implants, the mean Periotest value (PTV) of mandibular implants was -4.14 (anterior, -4.22; posterior, -4.06) versus -3.24 for maxillary implants (anterior, -2.91; posterior, -3.91). Implants in the densest bone (quality 1) had the lowest mean PTV (-4.13), followed by quality 2 (-4.00), quality 3 (-3.58), and quality 4 (-2.64). PMID- 9393429 TI - Changes in patient screening for a clinical study of dental implants after increased awareness of tobacco use as a risk factor. AB - Independent external monitoring committees are an important part of scientific clinical trials. They monitor patient safety, study progress, investigators' performance, and accurate interpretation/reporting of the study data. Data trends observed by a study monitoring committee detected a change in the pattern of patient screening by investigators after an increased awareness that tobacco use could directly compromise the osseointegration of root-form dental implants. This increased awareness is believed to have altered the number of active smokers accepted into a multicenter prospective dental implant study. Recent data analyses indicate that the success ratios were improved by alterations in this discretionary inclusion-exclusion criterion. PMID- 9393431 TI - Relationships between H-NS, sigma S, SpvR and growth phase in the control of spvR, the regulatory gene of the Salmonella plasmid virulence operon. AB - The sigma S-regulated gene spvR of Salmonella typhimurium encodes an autoregulatory protein required for transcriptional activation of the virulence operon spvABCD. A mutation in the histone-like protein H-NS, which negatively controls the sigma S level, has been reported to increase spv gene expression in S. typhimurium strain LT2. In agreement with this, we found that transcription of spvR and spvABCD was derepressed in hns strains of Escherichia coli and S. typhimurium. Moreover, levels of spv gene expression in hns rpoS double mutants were higher than expression levels in mutants deficient in rpoS alone, and were close to those measured in wild-type strains. This demonstrates that H-NS contributes to spv gene regulation independently of its function in controlling the sigma S level. Since the same start site was used for spvR gene transcription in wild-type as in hns and hns rpoS mutant strains, it is likely that the spvR promoter. spvRp1, can be recognized efficiently by an RNA polymerase containing sigma 70. The spvR promoter region shows an intrinsic DNA curvature that might be a determinant in H-NS- and/or sigma S-mediated control. A single amino acid substitution, Leu to Pro at position 265, abolished the regulatory function of SpvR in E. coli and Salmonella, implicating the C-terminal domain of SpvR in its structure and/or regulatory function. The spvR265 allele is not transcribed at detectable levels in hns or hns rpoS strains, suggesting that activation of spvRp1 in these strains remains dependent on SpvR. Thus, we propose a model for spvR gene regulation in which SpvR acts as a co-regulator of an RNA polymerase containing either sigma 70 (in the absence of H-NS) or sigma S, to induce transcriptional initiation at spvRp1. Moreover, growth-phase regulation of spv gene expression was maintained in hns and hns rpoS strains, indicating that an additional element, besides sigma S, is involved in the growth-phase regulation in rich medium. PMID- 9393430 TI - Success of multiple endosseous dental implant designs to second-stage surgery across study sites. AB - A multicenter clinical study of dental implants is being conducted by the Dental Implant Clinical Research Group to investigate the influence of implant design, application, and site on clinical performance and crestal bone. This article reports on the percentage of success up to implant uncovering for different implant designs and the distribution of failures across study sites. Data from 2,847 implants placed at 32 study sites were analyzed. Percentages of success up to implant uncovering were calculated for study implants overall, by implant design, by implant design within study strata, and according to individual study sites and quartile groupings of sites based on success. Comparisons were made, with chi-square and exact tests employed where appropriate. Differences were found between the different implant designs for the study overall, and between implant designs within the different study strata. Although some implant designs were found to have generally high success across study sites, some study sites designated as having surgeons with less experience tended to have higher failure levels, and one implant design failed at higher rates in a subset of study sites. The percentage and distribution of implant failures varied across study sites and by implant design. These differences appeared to be in part related to the level of experience of the surgeons. Further investigation should focus on identification of factors that contribute to higher success in implant placement with different implant designs. PMID- 9393432 TI - Transposition-mediated transcriptional overexpression as a mechanism of insecticide resistance. AB - It has been proposed that amplification of genes for esterase that provide resistance to insecticides may originate from transposition events. To test this hypothesis, we have constructed a minigene coding for a soluble acetylcholinesterase under the control of a nontissue-specific promoter (hsp70). When introduced into Drosophila, the gene is expressed in all tissues and the extra acetylcholinesterase produced confers a low level of insecticide resistance (twofold). The minigene was mobilized by crossing the initial transformant with a strain providing a source of P-element transposase. After 34 generations of exposure to the organophosphate parathion, we obtained a strain with a higher resistance (fivefold). This strain had only one extra Ace gene, which overexpressed acetylcholinesterase. Thus, following transposition, resistance resulted from the overexpression of a single copy of the gene and not from gene amplification. PMID- 9393433 TI - Cloning of the gene encoding the catalytic subunit of casein kinase II from the yeast Yarrowia lipolytica. AB - Casein kinase II from the yeast Yarrowia lipolytica is a heterotetramer of the form alpha alpha' beta 2. We report on the cloning and sequencing of a partial cDNA and of the complete genomic DNA coding for the catalytic alpha subunit of the casein kinase II from this yeast species. The sequence of the gene coding for this enzyme has been analyzed. No intron was found in the gene, which is present in a single copy. The deduced amino acid sequence of the gene shows high similarity with those of alpha subunit described in other species, although, uniquely, Y. lipolytica CKII alpha lacks cysteines. We find that the alpha subunit sequence of Y. lipolytica CKII is shown greater homology with the corresponding protein from S. pombe than with that from S. cerevisiae. We have analyzed CKII alpha expression and CKII alpha activity. We show that expression of this enzyme is regulated. The catalytic subunit is translated from a single mRNA, and the enzyme is present at a very low level in Y. lipolytica, as in other yeasts. PMID- 9393434 TI - Mutational analysis of Cak1p, an essential protein kinase that regulates cell cycle progression. AB - In Saccharomyces cerevisiae, entry into S phase requires the activation of the protein kinase Cdc28p through binding with cyclin Clb5p or Clb6p, as well as the destruction of the cyclin-dependent kinase inhibitor Sic1p. Mutants that are defective in this activation event arrest after START, with unreplicated DNA and multiple, elongated buds. These mutants include cells defective in CDC4, CDC34 or CDC53, as well as cells that have lost all CLB function. Here we describe mutations in another gene, CAK1, that lead to a similar arrest. Cells that are defective in CAK1 are inviable and arrest with a single nucleus and multiple, elongated buds. CAK1 encodes a protein kinase most closely related to the Cdc2p family of protein kinases. Mutations that lead to the production of an inactive kinase that can neither autophosphorylate, nor phosphorylate Cdc28p in vitro are also incapable of rescuing a cell with a deletion of CAK1. These results underscore the importance of the Cak1p protein kinase activity in cell cycle progression. PMID- 9393435 TI - Identification of the Saccharomyces cerevisiae genes STB1-STB5 encoding Sin3p binding proteins. AB - The yeast SIN3 gene functions as a transcriptional repressor, despite the fact that Sin3p does not bind DNA directly. We have conducted a two-hybrid screen to look for proteins that interact with Sin3p, using the PAH2 domain of Sin3p as bait. Five new genes, STB1-STB5 were identified, as well as the STB6 gene, which is similar to STB2. STB1, STB2, STB3, and STB6 are novel genes, and STB4 and STB5 encode C6 zinc cluster DNA-binding proteins. None of these genes is essential for viability, and several of these genes may encode transcriptional activators. Several special problems were encountered in using a transcriptional repressor in a two-hybrid screen. For example, the STB genes will interact with a LexA Sin3(PAH2) fusion protein containing a region of Sin3p, but a LexA-Sin3p fusion protein containing full-length Sin3p, along with a STB clone, does not produce two-hybrid activation of a transcriptional reporter. In addition, a sin3 mutation reduces the transcriptional activation by two-hybrid partners, suggesting that a sin3 mutation reduces the transcriptional efficiency of the Gal4p and VP16 activation domains. We have shown previously that Sin3p is part of a large multiprotein complex, and we show here that Stb1p and Stb2p are present in this complex. PMID- 9393436 TI - Transposon-like structure of a new plasmid-encoded restriction-modification system in Rhizobium leguminosarum VF39SM. AB - Total DNA isolated from Rhizobium leguminosarum VF39SM cells is resistant to cleavage by the restriction endonuclease PstI. Plasmid curing and transfer studies localized this phenotype to pRleVF39b, the second smallest of six plasmids found in this bacterium. In vitro selection for vector modification was employed to isolate a presumptive methylase gene (M.Rle39BI) from a plasmid gene library. Total and plasmid DNAs isolated from E. coli containing M.RleBI were resistant to digestion by PstI. Sequence data suggested that a putative restriction endonuclease (R.Rle39BI) was also encoded on the same fragment. The two genes were flanked by identical copies of a putative insertion sequence, which was also present in several copies elsewhere in the VF39SM genome. The presence of this element in other strains examined suggested that this element is indeed an insertion sequence. The differences in G/C content between the DNA coding for the R/M system and that of the IS element suggest that this DNA region may have been acquired by horizontal transfer. PMID- 9393437 TI - Clustered amino acid substitutions in the yeast transcription regulator Pdr3p increase pleiotropic drug resistance and identify a new central regulatory domain. AB - In the yeast Saccharomyces cerevisiae mutations in the genes encoding the transcription factors Pdr1p and Pdr3p are known to be associated with pleiotropic drug resistance mediated by the overexpression of the efflux pumps Pdr5p, Snq2p, and Yor1p. Mutagenesis of PDR3 was used to induce multidrug resistance phenotypes and independent pdr3 mutants were isolated and characterized. DNA sequence analysis revealed seven different pdr3 alleles with mutations in the N-terminal region of PDR3. The pdr3 mutants were semidominant and conferred different drug resistance patterns on host strains deleted either for PDR3 or for PDR3 and PDR1. Transactivation experiments proved that the mutated forms of Pdr3p induced increased activation of the PDR3, PDR5, and SNQ2 promoters. The amino acid changes encoded by five pdr3 mutant alleles were found to occur in a short protein segment (amino acids 252-280), thus revealing a regulatory domain. This region may play an important role in protein-DNA or protein-protein interactions during activation by Pdr3p. Moreover, this hot spot for gain-of-function mutations overlaps two structural motifs, MI and MII, recently proposed to be conserved in the large family of Zn2Cys6 transcription factors. PMID- 9393438 TI - Molecular and phenotypic characterization of yeast PDR1 mutants that show hyperactive transcription of various ABC multidrug transporter genes. AB - Mutations at the yeast PDR1 transcriptional regulator locus are responsible for overexpression of the three ABC transporter genes PDR5, SNQ2 and YOR1, associated with the appearance of multiple drug resistance. The nucleotide sequences of 13 alleles of PDR1, comprising 6 multidrug resistance mutants, 1 intragenic suppressor and 6 wild types, have been determined. Single amino acid substitutions were shown to result from the mutations pdr1-2 (M308I), pdr1-3 (F815S), pdr1-6 (K302Q), pdr1-7 (P298A) and pdr1-8 (L1036 W), whereas the intragenic suppressor mutant pdr1-100 is deleted for the two amino acids L537 and A538. An isogenic series of strains was constructed containing the mutant alleles pdr1-3, pdr1-6 and pdr1-8 integrated into the genome. We found that the levels of resistance to cycloheximide, oligomycin, 4-nitroquinoline-N-oxide and ketoconazole were increased in all three mutants. The increase was more pronounced in the pdr1-3 than in the pdr1-6 and pdr1-8 mutants. Studies of the activity of the promoters of the ABC genes PDR5, SNQ2 and YOR1 demonstrated that the combination of the PDR5 promoter and the pdr1-3 mutation resulted in the highest level of promoter induction. Concomitantly, the level of PDR5 mRNA, of Pdr5p protein, and of its associated nucleoside triphosphatase activity, was strongly increased in the plasma membranes of the PDR1 mutants. Again, the pdr1-3 allele was associated with a stronger effect than the pdr1-8 and pdr1-6 alleles. The locations of the mutations in the PDR1 gene indicate that at least three different regions distributed throughout the Pdr1p transcription factor may be mutated to generate a Pdr1p with considerably increased transcriptional activation potency. These gain-of-function mutations support the concept, recently proposed, that in members of the large family of yeast Zn2Cys6 transcription factors a central inhibitory domain exists (delineated by the pdr1 7, pdr1-6 and pdr1-2 mutations). This domain may interact in a locked conformation with a putative, more C-terminally located inhibitory domain (mutated in pdr1-3), and with the putative activation domain (mutated in pdr1-8). PMID- 9393439 TI - SigX of Bacillus subtilis replaces the ECF sigma factor fecI of Escherichia coli and is inhibited by RsiX. AB - Analysis of the Bacillus subtilis genome sequence revealed two open reading frames, designated sigX and ypuN (now termed rsiX), that are homologous to fecI and fecR, respectively, of Escherichia coli. fecI encodes a sigma 70-type factor that is necessary for transcription of the ferric citrate transport genes fecABCDE. fecR encodes a cytoplasmic transmembrane protein that is required for the induction of fec transport gene transcription by ferric citrate binding to the FecA outer membrane receptor protein. Investigation of the SigX and RsiX activities disclosed that they are not involved in ferric citrate utilization- since ferric citrate did not serve as an iron source for B. subtilis SG64--or in the regulation of any other ferric siderophore transport system tested. Strains deleted for sigX or rsiX displayed no phenotype under aerobic or anoxic conditions. However, cloned sigX complemented an E. coli fecI mutant, and the Fur box upstream of sigX responded to the E. coli iron regulatory protein Fur. The purified SigX protein was required for in vitro transcription of a sigX containing DNA fragment by the E. coli RNA polymerase core enzyme. Autoregulation of sigX was also found in vivo using a sigX'-lacZ gene fusion. RsiX inhibited SigX activity in vivo and in vitro and stabilized the SigX protein. RsiX was localized in the membrane fraction. When RsiX is present, SigX is found in the membrane fraction; in the absence of RsiX, some SigX is detectable in the cytoplasm. We conclude that SigX is a sigma factor that belongs to the ECF (extracytoplasmic function) sigma 70-factor family. It is not known which promoters are recognized by SigX in B. subtilis. SigX may be involved in the regulation of iron metabolism, as evidenced by its activity in E. coli. PMID- 9393440 TI - Fil1, a G-protein alpha-subunit that acts upstream of cAMP and is essential for dimorphic switching in haploid cells of Ustilago hordei. AB - A constitutive mutation, fil1, that causes filamentous growth in the haplophase of the dimorphic smut fungus Ustilago hordei, was previously shown to be genetically associated with a 50-kb deletion within a 940-kb chromosome. Physiological studies suggested that a gene that functions upstream of adenylyl cyclase was deleted in the mutant. Representational difference analysis of isolated chromosomes was used to obtain deletion-specific DNA probes and corresponding genomic cosmid clones. Complementation analysis identified a cosmid clone and subsequently a 2.1-kb insert that converted transformants of the mutant strain 10.1a(fil1) from the filamentous to the sporidial cell type. A single open reading frame of 354 codons that encodes a putative alpha-subunit of the heterotrimeric G-proteins was identified. Fil1 displayed a high degree of sequence identity to Gpa1 from the basidiomycete Cryptococcus neoformans and CPG 2 from the ascomycete Cryphonectria parasitica. FIL1, when introduced on a self replicating vector, was found to suppress filamentous growth of starved haploid wild-type strains and restore normal mating response to the fil1 mutant, but did not suppress sexual dimorphism of either strain. Fil1 appears to function analogously to mammalian G alpha proteins, which are coupled to cAMP production via adenylyl cyclase, to regulate dimorphic switching in U. hordei. PMID- 9393441 TI - Genetic and molecular characterization of Neurospora crassa mus-23: a gene involved in recombinational repair. AB - A newly isolated mutant, mus-23, of Neurospora crassa was found to be highly sensitive to a wide variety of mutagens, including UV light, methyl methanesulfonate, 4-nitroquinoline 1-oxide, N-methyl-N'-nitro-N-nitrosoguanidine and tert-butyl hydroperoxide. This mutant was originally isolated as a mutant that could not grow on medium containing histidine. Meiosis and sporulation were defective in homozygous crosses between mus-23 haploids. The mus-23 gene is located on the right arm of LGII, between fl and trp-3. Analyses of epistasis between mus-23 and other mutations that cause defects in DNA repair indicated that the mus-23 gene belongs to the same DNA repair group as mei-3, which is the Neurospora homolog of the Saccharomyces cerevisiae gene RAD51. The double mutant carrying mus-23 and uvs-3 mutations was lethal. The mus-23 gene was cloned by complementation of the MMS-sensitive phenotype of the mus-23 mutant. The gene contained an open reading frame of 1578 bp and did not contain any introns. The molecular weight of the predicted mus-23 gene product was 60.4 kDa. Computer analyses revealed that the MUS23 protein has significant homology to Mre11p, which is known to be involved in recombinational repair in S. cerevisiae. The level of mus-23 transcripts increased significantly within 60 min of treatment with UV or MMS and then gradually decreased. The role of MUS23 protein in recombinational repair is discussed. PMID- 9393442 TI - Isolation and analysis of functional homologues of the secretion-related SAR1 gene of Saccharomyces cerevisiae from Aspergillus niger and Trichoderma reesei. AB - The Aspergillus niger and Trichoderma reesei genes encoding the functional homologues of the small GTP-binding protein SAR1p, which is involved in the secretion pathway in Saccharomyces cerevisiae, have been cloned and characterised. The A. niger gene (sarA) contains five introns, whereas the T. reesei gene (sar1) has only four. In both cases the first intron is at the same position as the single S. cerevisiae SAR1 intron. The encoded proteins show 70 80% identity to the SAR1 protein. Complementation of S. cerevisiae sar1 and sec12 mutants by expression vectors carrying the A. niger sarA and T. reesei sar1 cDNA clones confirmed that the cloned genes are functional homologues of the S. cerevisiae SAR1 gene. Three mutant alleles of the A. niger sarA gene (D29G, E109K, D29G/E109K), generated by site-directed mutagenesis, revealed a thermosensitive dominant-negative phenotype in the presence of the wild-type sarA allele. This result contrasts with the situation in S. cerevisiae, where similar mutations have a thermosensitive phenotype. Taken together, our results indicate that the sarA gene is involved in an essential function in A. niger. PMID- 9393443 TI - Gene activation at a distance and telomeric silencing are not affected by yeast histone H1. AB - Until recently, it was believed that the budding yeast Saccharomyces cerevisiae has no histone H1 gene. However, a search of the yeast genome database revealed a possible H1 homologue of 258 amino acids, termed yeast histone H1 (HHO1). The protein shows 36% identity to the human H1 core domain over a stretch of 93 amino acids. Unlike other H1 proteins, Hho1p has a second possible core domain which shows 43% identity to the first core domain. Since vertebrate H1 histone had been implied in gene repression as well as gene activation at a distance, we tested the effect of deleting the yeast H1-like gene on remote activation of a modified GAL1 promoter, which contains a synthetic GAL4 binding site close to the TATA box, and the natural UASG, consisting of four GAL4 binding sites. Different spacing up to 1.8 kb between the proximal binding site and the distal UASG enhancer revealed no differences in gene activation between wild-type and knockout strains. Overexpression of a heterologous histone H1 from sea urchin showed an overall inhibition of gene activation by the GAL1 promoter, whereas overexpression of the yeast histone H1 had no effect. Also, the expression of A1, ALPHA2 or SUC2 genes, all of which are known to be responsive to an altered chromatin structure, was unchanged in HHO1 knockout or HHO1-overexpressing strains when compared to wild-type cells. We also considered the possibility that HHO1 was involved in forming the heterochromatin at telomeres. On testing for telomeric silencing of a URA reporter gene introduced 1.3 kb away from the chromosomal end, we again observed no differences between wild-type and knockout strains. Thus, the yeast histone H1-like gene appears to have no role in gene activation at a distance or in silencing under the conditions tested. It remains to be seen whether the yeast H1 histone is a gene-specific regulator rather than a general chromatin-associated protein. PMID- 9393444 TI - Antisense suppression of the putative ribosomal protein S3A gene disrupts ovarian development in Drosophila melanogaster. AB - The Drosophila melanogaster homologue of the Anopheles gambiae C3 cDNA has been isolated and characterized by sequence analysis. The encoded protein was localized by immunochemical and immunocytochemical methods. The Drosophila C3 protein is highly similar to homologues of disputed function, which have previously been identified in fungi, plants and animals. The protein is ubiquitous and localized in the cytoplasm. Cell fractionation followed by detection with a specific antibody preparation shows that the protein is associated with the 40S ribosomal subunit. The C3 gene is located in section 101F of chromosome 4. Antisense transgenic analysis shows that this gene is essential for oogenesis. The most prominent phenotype resulting from antisense depletion of C3 RNA is disappearance of the follicular cells of the ovary (where the concentration of C3 protein is normally high) and abnormalities of the associated germline derivatives, leading to failure of egg production. PMID- 9393445 TI - Differential activation of two ACC oxidase gene promoters from melon during plant development and in response to pathogen attack. AB - ACC (1-aminocyclopropane-1-carboxylate) oxidase genes are differentially expressed in melon during development and in response to various stresses. We investigated the molecular basis of their transcription by analyzing the 5' untranslated regions of the ACC oxidase genes CM-ACO1 and CM-ACO3. In order to determine how their temporal and spatial expression patterns were established, we fused the promoter regions of CM-ACO1 (726 bp) and CM-ACO3 (2260 bp) to the beta glucuronidase (GUS) reporter gene and examined their regulation in transgenic tobacco plants. The CM-ACO1 promoter was able to drive GUS expression in response to wounding, and to treatment with ethylene or copper sulfate. It was also rapidly induced (8-12 h postinoculation) in tobacco leaves inoculated with the hypersensitive response (HR)-inducing bacterium Ralstonia solanacearum. Expression was also observed during compatible interactions but was delayed. In contrast, the CM-ACO3 promoter was not expressed in response to infection, but was up-regulated during flower development. Both promoters were regulated during leaf senescence but in different patterns. The CM-ACO1-driven GUS activity increased sharply concomitantly with the onset of chlorophyll breakdown, while the CM-ACO3 promoter drove strong GUS expression in green, fully expanded leaves and this declined at the onset of senescence. This result is consistent with the expression patterns of these two genes in senescent melon leaves. These data suggest that the regulation of expression of CM-ACO1 is related preferentially to stress responses, whereas CM-ACO3 seems to be associated with developmental processes. The possible role of ethylene is discussed, particularly in the regulation of the CM-ACO1 gene in response to stress and during senescence. PMID- 9393446 TI - Dap (Defective aleurone pigmentation) mutations affect maize aleurone development. AB - Dap (Defective aleurone pigmentation) is the designation for mutations in maize that give rise to a characteristic dappled endosperm phenotype, consisting of patches of purple tissue, of variable size and shape, on a yellow background. Features shared by all Dap mutants are: dominant expression when they are maternally derived, lack of expression or transmission when they originate from pollen, failure to recover homozygous Dap genotypes, reduced frequency of Dap seeds in the progeny of outcrosses of Dap/+ females, association of the dappled phenotype with reduction in seed size. The mutants so far tested, six in all, can be grouped into two classes, one including male-transmissible (MT) isolates not expressed in the endosperm if their contribution is paternal, and a second class of isolates (NMT) that are permanently lost following paternal transmission. We suggest that the NMT mutations are on a chromosome that carries an intercalary deletion. Assuming linkage between the mutant and the deletion, selection against the deficient chromosome during male gametogenesis would account for the failure to recover Dap seeds in the progeny of Dap/+ male parents. We have obtained genetic evidence supporting this hypothesis. This interpretation, however, does not apply to MT alleles. For these, other mechanisms, such as imprinting and/or dosage effects may be proposed. The mutable pattern in the endosperm to which all Dap mutants give rise is an intriguing phenotype which remains to be clarified. An unexpected finding is that aleuronic and subaleuronic cells corresponding to the colourless areas are abnormal in shape and anthocyanin biosynthesis is blocked in these cells. This finding calls for further investigation in light of a possible connection between flavonoid precursors and cell shape. PMID- 9393447 TI - Genetic analyses of the formation of the serrated margin of leaf blades in Arabidopsis: combination of a mutational analysis of leaf morphogenesis with the characterization of a specific marker gene expressed in hydathodes and stipules. AB - Developmental control of the formation of the serrated margin of leaf blades was investigated. First, the expression was characterized of a marker gene encoding beta-glucuronidase in strain #1-35-38, a transgenic strain of Arabidopsis thaliana (L.) Heynh, derived by the use of a previously described transposon tagging system. In strain #1-35-38, expression of the marker gene was tissue specific, being restricted to stipules and the toothed margins of laminae. Using this transgenic marker gene, we examined the development of leaf blade margins in Arabidopsis. We compared the pattern of expression of the marker gene in the leaves of the wild-type plant with that in plants carrying the asymmetric leaves1 (as1) mutation, which causes dramatic changes in leaf-blade morphology in Arabidopsis. The as1 mutant showed normal morphology of early leaf primordia. The mutation affected the development of leaf segmentation in Arabidopsis without any change in the number or morphology of cells in laminae. The as1 mutation affected leaf morphology independently of mutations in other genes known to affect leaf morphogenesis, such as the acaulis1 mutation and the angustifolia mutation. Based upon these results, the development of the morphology of leaf margins in Arabidopsis is discussed. PMID- 9393448 TI - Analysis of seven genes from the eryAI-eryK region of the erythromycin biosynthetic gene cluster in Saccharopolyspora erythraea. AB - The gene cluster (ery) governing the biosynthesis of the macrolide antibiotic erythromycin A by Saccharopolyspora erythraea contains, in addition to the eryA genes encoding the polyketide synthase, two regions containing genes for later steps in the pathway. The region 5' of eryA, and lying between eryA and the gene eryK, which is known to encode the C-12 hydroxylase, has been sequenced and shown to contain seven additional open reading frames (ORFs 13-19). On the basis of sequence similarities, roles are proposed for several of these ORFs in the biosynthesis of the deoxysugar mycarose and the deoxyaminosugar desosamine. A chromosomal mutant carrying a deletion in ORF15 has been constructed and shown to accumulate 3-O-mycarosylerythronolide B, as expected for an eryC mutant. Similarly, a chromosomal mutant carrying a deletion in ORF16 has been constructed and shown to accumulate erythronolide B, as expected for an eryB mutant. PMID- 9393449 TI - A germline restricted, highly repetitive DNA sequence in Paramyxine atami: an interspecifically conserved, but somatically eliminated, element. AB - In some species of hagfish, the phenomenon of chromosome elimination occurs during embryogenesis. However, only two repetitive DNA families are known to be represented in chromosomes that are eliminated from somatic cells of the Japanese hagfish Eptatretus okinoseanus. Using molecular analyses, another germ line restricted, highly repetitive DNA family has been detected in another Japanese hagfish, Paramyxine atami. The repeat unit of this family, which is 83 bp long, has been designated "EEPa1", for Eliminated Element of P. atami 1. DNA filter hybridization using EEPa1 as a probe revealed that this family is shared among several species and is conserved in the germline DNA. Although eliminated, repetitive DNA that is shared interspecifically has not been reported in hagfish species, cases of chromatin diminution and chromosome elimination processes have been described previously in other organisms. The patterns and intensities of hybridization signals suggest that members of the repetitive DNA family defined by EEPa1 have undergone concerted molecular evolution. PMID- 9393450 TI - Three members of the S multigene family are linked to the S locus of Brassica. AB - Two self-incompatibility genes in Brassica, SLG and SRK (SLG encodes a glycoprotein; SRK encodes a receptor-like kinase), are included in the S multigene family. Products of members of the S multigene family have an SLG-like domain (S domain) in common, which may function as a receptor. In this study, three clustered members of the S multigene family, BcRK1, BcRL1 and BcSL1, were characterized. BcRK1 is a putative functional receptor kinase gene expressed in leaves, flower buds and stigmas, while BcRL1 and BcSL1 are considered to be pseudogenes because deletions causing frameshifts were identified in these sequences. Sequence and expression pattern of BcRK1 were most similar to those of the Arabidopsis receptor-like kinase gene ARK1, indicating that BcRK1 might have a function similar to that of ARK1, in processes such as cell expansion or plant growth. Interestingly, the region containing BcRK1, BcRL1 and BcSL1 is genetically linked to the S locus and the physical distance between SLG, SRK and the three S-related genes was estimated to be less than 610 kb. Thus the genes associated with self-incompatibility exist within a cluster of S-like genes in the genome of Brassica. PMID- 9393451 TI - Ectopic anthocyanin pigmentation in maize as a tool for defining interactions between homologous regulatory factors. AB - The duplicated R and Sn genes are involved in the regulation of the maize anthocyanin biosynthetic pathway, encoding tissue-specific products that are homologous to the helix-loop-helix transcriptional activators. Sn determines the pigmentation of the mesocotyl, leaf basis and pericarp, while R determines pigmentation in various tissues, but not in the mesocotyl. In the progeny derived from test-crosses of R/Sn heterozygous plants, a high frequency of R plants exhibiting mesocotyl pigmentation was observed; these derivatives were defined as R*. In R* plants, the presence of this novel trait was not accompanied by the acquisition of Sn or by gross DNA rearrangements in the R profile. Accordingly, RT-PCR analysis showed that mesocotyl pigmentation in R* was attributable to the resident R gene. The occurrence of R* was observed with all R alleles tested, and was enhanced when a P component was present. The heritability of R* was shown only in the case of the standard R-r allele, which carries a functional P component. In addition, we observed that R* can influence other R alleles, transferring the ability to pigment the mesocotyl. R* is unstable, showing a tendency to return to its original state after a few generations. In R* plants there was a correlation between observed ectopic pigmentation and an increase in the level of A1 transcript but, surprisingly, not in the accumulation of R transcript. The results obtained from the analysis of test crosses of rSn/r delta plants suggest that an unlinked genetic factor accounts for the ectopic pigmentation. Concomitant occurrence of epigenetic events might explain the observed instability and reversibility noted above. Further study of this phenomenon might help to elucidate the basis of the interaction between homologous and non-homologous regulators. PMID- 9393452 TI - Genetic analysis of mutagenesis in aging Escherichia coli colonies. AB - Bacteria live in unstructured and structured environments, experiencing feast and famine lifestyles. Bacterial colonies can be viewed as model structured environments. SOS induction and mutagenesis have been observed in aging Escherichia coli colonies, in the absence of exogenous sources of DNA damage. This cAMP-dependent mutagenesis occurring in Resting Organisms in a Structured Environment (ROSE) is unaffected by a umuC mutation and therefore differs from both targeted UV mutagenesis and recA730 (SOS constitutive) untargeted mutagenesis. As a recB mutation has only a minor effect on ROSE mutagenesis it also differs from both adaptive reversion of the lacI33 allele and from iSDR (inducible Stable DNA Replication) mutagenesis. Besides its recA and lexA dependence, ROSE mutagenesis is also uvrB and polA dependent. These genetic requirements are reminiscent of the untargeted mutagenesis in lambda phage observed when unirradiated lambda infects UV-irradiated E. coli. These mutations, which are not observed in aging liquid cultures, accumulate linearly with the age of the colonies. ROSE mutagenesis might offer a good model for bacterial mutagenesis in structured environments such as biofilms and for mutagenesis of quiescent eukaryotic cells. PMID- 9393453 TI - Role of Escherichia coli cspA promoter sequences and adaptation of translational apparatus in the cold shock response. AB - A shift in growth temperature from 37 degrees C to 15 degrees C leads to a dramatic increase in the level of CspA, the major cold shock protein of Escherichia coli. To investigate the molecular basis of this induction, we considered the relevance of transcriptional and posttranscriptional controls by analyzing the steady-state levels of transcripts and the expression of reporter genes in cells carrying a set of cspA promoter fragments of variable length fused to lacZ or cat genes. We demonstrate that: (i) the core cspA promoter (from -40 to +16) responds to cold shock and a mutation at -36 increases the relative activity of the promoter at low temperature by threefold; (ii) the sequences upstream of -40 have a positive effect on expression at 37 degrees C, but no effect on the cold shock response; (iii) by virtue of their influence on mRNA stability, the downstream sequences (from +81 to +165) reduce expression at 37 degrees C and increase the intensity of the cold shock response; (iv) mutations in the GCACATCA and CCAAT motifs, present at +1/-4 and between the -10 and -35 elements, respectively, do not affect the cold shock response of the cspA promoter; (v) following cold shock, a modification of the protein synthetic machinery takes place that allows preferential translation of cspA mRNA relative to the non-cold shock cat and lacZ mRNAs. The quantitatively modest transcriptional activation shown by the core promoter of cspA following cold shock suggests that transcriptional activation can significantly contribute to cold shock induction only when coupled to posttranscriptional controls, such as alterations in mRNA stability and the translational apparatus. PMID- 9393454 TI - Genetic basis of the MbrC "ploidy" phenotype in Escherichia coli. AB - The mbrC17 mutation in Escherichia coli had been shown to cause conditional growth defects and an increase in the quantity of DNA per cell. The present work was aimed at identifying the mutation. Sequencing showed that the MbrC17 phenotype does not involve glr (murI), as previously suggested. P1 transduction data indicated that the mbrC17 mutation is closely linked to rpoB, and allele exchange showed it to lie within the secE-nusG operon. A single change relative to wild type was found in the secE-nusG region from the mbrC17 strain, a G-->A mutation 23 bp upstream of the secE coding sequence. This mutation causes a two fold increase in the concentration of secE-nusG mRNA. PMID- 9393455 TI - Allelic variation at a hypervariable compound microsatellite locus in the ascomycete Ascochyta rabiei. AB - The genome of the fungal chickpea pathogen Ascochyta rabiei was screened for polymorphisms by microsatellite-primed PCR. While ethidium-bromide staining of electrophoretically separated amplification products showed only limited polymorphism among 24 Tunisian A. rabiei isolates, Southern hybridization of purified PCR fragments to restriction digests of fungal DNA revealed polymorphic DNA fingerprints. One particular probe that gave rise to a hypervariable single locus hybridization signal was cloned from the Syrian isolate AA6 and sequenced. It contained a large compound microsatellite harbouring the penta- and decameric repeat units (CATTT)n, (CATTA)n, (CATATC-ATTT)n and (TATTT)n. We call this locus ArMS1 (Ascochyta rabiei microsatellite 1). Unique flanking sequences were used to design primer pairs for locus-specific microsatellite amplification and direct sequencing of additional ArMS1 alleles from Tunisian and Pakistani isolates. A high level of sequence variation was observed, suggesting that multiple mutational mechanisms have contribute to polymorphism. Hybridization and PCR analyses were performed on the parents and 62 monoascosporic F1 progeny derived from a cross between two different mating types of the fungus. Progeny alleles could be traced back to the parents, with one notable exception, where a longer than expected fragment was observed. Direct sequencing of this new length allele revealed an alteration in the copy number of the TATTT repeat [(TATTT)53 to (TATTT)65], while the remainder of the sequence was unchanged. PMID- 9393456 TI - In vivo and in vitro studies on interactions between the components of the hemolysin (HlyA) secretion machinery of Escherichia coli. AB - The glycopeptide antibiotic vancomycin blocks cell wall synthesis in Escherichia coli only when it can reach its target site in the periplasm. In vivo, sensitivity to vancomycin is enhanced in the presence of the hemolysin (hly) determinant of E. coli or its translocator portion hlyBD. Two different mutations in hlyD alter the cell's susceptibility to vancomycin: mutations in the tolC homologous region of hlyD increase vancomycin resistance, whereas mutations at the 3'-terminus of hlyD lead to hypersensitivity to vancomycin and to the accumulation of large periplasmic and cytoplasmic pools of this antibiotic in E. coli. These effects are only observed in the presence of functional HlyB and TolC, the two other components of the hemolysin secretion machinery. A defect in TolC causes hyperresistance to vancomycin, even when present together with a mutant HlyD protein which in the presence of TolC renders E. coli hypersensitive to vancomycin. Lipid bilayer experiments in vitro revealed specific interactions between TolC and vancomycin or HlyD protein. Second-site suppressor mutations in hlyD and hlyB were obtained, which abolish the hypersensitive phenotype caused by the 3'-terminal mutations in hlyD. Our results are compatible with the idea that (a) TolC, together with the TolC-homologous part of HlyD, forms a pore in the outer membrane through which hemolysin is released and vancomycin taken up; and (b) the C-terminal sequence of HlyD interacts with periplasmic loop(s) of HlyB to form a closed channel spanning the periplasm. PMID- 9393457 TI - Repression of beta-actin synthesis and persistence of ribosomal protein synthesis after infection of HeLa cells by herpes simplex virus type 1 infection are under translational control. AB - Synthesis and assembly of ribosomal proteins into mature ribosomes persist late after infection of cells with herpes simplex virus type 1, while synthesis of beta-actin is drastically shut off. Since mRNAs encoding ribosomal proteins and beta-actin undergo concomitant degradation in infected HeLa cells, we have advanced the hypothesis that translation of the remaining mRNAs is differentially controlled after infection. The behaviour of mRNAs for three ribosomal proteins and for beta-actin was investigated during the course of infection. In uninfected cells, beta-actin mRNAs are associated with large polyribosomes, while only a part of ribosomal protein mRNAs are present in polyribosomes. In the course of infection, beta-actin mRNAs are released from the ribosomes and are sequestered with 40S ribosomal subunits. Simultaneously, ribosomal protein mRNAs become associated with an increased number of ribosomes, even late in infection. In addition, virally induced phosphorylation of ribosomal protein S6 is more efficient in pre-existing ribosomes than in newly assembled ribosomes. These results indicate that in infected cells (i) translation of beta-actin mRNA is selectively inhibited at a step necessary for binding the 60S ribosomal subunits; (ii) the rate of initiation of translation of ribosomal protein mRNAs increases after infection; and (iii) it is likely that translation of ribosomal protein mRNAs takes place preferentially on pre-existing ribosomes. PMID- 9393458 TI - Temperature regulates expression of the Drosophila vestigial gene only in mutant wing discs. AB - All Vestigial mutants in Drosophila melanogaster display a thermosensitive phenotype, with the exception of two which disrupt an intronic wing-specific enhancer element. Here we report a very unusual transcriptional regulation; temperature changes are associated with alterations in the level of vg expression only in the wing disc of thermosensitive mutant flies and not in the brain. No effect is observed in the wild-type strain. The tissue specificity of the temperature effect indicates an involvement of the intronic wing-specific enhancer element in determining the thermosensitivity of mutants. PMID- 9393459 TI - Distraction osteogenesis for lengthening of the hard palate: Part I. A possible new treatment concept for velopharyngeal incompetence. Experimental study in dogs. AB - Many procedures have been described to correct velopharyngeal incompetence. Significant complications can occur, and the results may not be satisfactory. If the short soft palate has satisfactory muscle function and if it could be moved toward the posterior pharyngeal wall by distraction osteogenesis of the hard palate, an entirely new concept of treatment for velopharyngeal incompetence would be available. The object of the present study was to explore the possibility of osteogenesis occurring in the hard palate in dogs after gradual distraction (callus distraction). Six adult, mix-bred dogs were anesthetized, and the palatal mucosa was elevated. A midpalatal transverse osteotomy and two lateral osteotomies were performed. Tantalum bone markers for cephalometric analysis were placed, and an individually fabricated, orthodontic-like distraction device with an expansion screw in the sagittal direction was inserted. The device was stabilized on the premolars and fixed to the palatal bone with titanium miniscrews. Gradual distraction began after a latency period of 10 to 18 days. The rate of the distraction varied from 0.25 to 0.75 mm per day. The device was left in place for 6 to 8 weeks after expansion to allow for bony consolidation. Assessment was by direct examination, cephalograms, computed tomography, and histology with bone labeling. Impressions of the jaws were taken preoperatively and after device removal to examine plaster cast changes in the dental occlusion. Cephalometric and computed tomographic scan analysis demonstrated a distraction of up to 8 mm. All gaps were filled with de novo osteogenesis. Comparison of the plaster casts revealed no change in the occlusion. At 1 month after distraction, the computed tomographic scan showed the first signs of ossification of the experimental gap from the anterior and posterior bone ends. After 4.5 months ossification was almost complete with a small translucent zone in the middle of the experimental gap. After 7 months ossification was complete. PMID- 9393460 TI - Distraction osteogenesis for lengthening of the hard palate: Part II. Histological study of the hard and soft palate after distraction. AB - To correct velopharyngeal incompetence, a new treatment concept was proposed in Distraction Osteogenesis for Lengthening of the Hard Palate: Part I (using lengthening of the hard and soft palate by distraction osteogenesis). Cephalometry and computed tomography showed successful elongation of the posterior hard palate with gradual calcification. Here the sequential use of fluorochrome markers (oxytetracycline, xylenol orange, DCAF [2,4-bis-N-N' dicarboxymethyl aminomethyl fluorescein], and alizarin complexone) during the distraction and retention period is reported together with the histologic investigations using light and laser scan microscopy without prior demineralization. The experimental gap showed de novo osteogenesis in all dogs. The new bone was always in continuity with the original anterior and posterior palatal bone margins. It either bridged the experimental gap fully or left a small central zone of fibrous tissue, in which eventual ossification occurred. Several distinct zones could be distinguished: A small central zone was found with parallel strains of collagen fibers, oriented longitudinally in the direction of the distraction. Next to this zone a layer of undifferentiated mesenchymal precursor cells was seen in direct contact to newly formed bone. The next zone was coarse woven bone, showing a transition to mature lamellar bone through remodeling. No evidence of endochondral bone formation was found, i.e., all dogs showed exclusively intramembranous bone formation. The soft tissues showed no signs of alteration: in particular, there was no necrosis or scar formation. The soft tissues were not thinned but appeared to have followed the longitudinal displacement. In conclusion, gradual distraction osteogenesis of the hard palate could be a possible method for lengthening the palate to treat velopharyngeal incompetence. PMID- 9393461 TI - Perception of postpalatoplasty speech differences in school-age children by parents, teachers, and professional speech pathologists. AB - The aims of this study were twofold: (1) to test the ability of parents and teachers to discriminate the speech of children with repaired cleft palate from that of their unaffected peers and (2) to compare these lay assessments of speech acceptability with the critical perceptual assessments of expert clinicians. The subjects for this study were 20 children of school age (age range, 8 to 12 years) who were drawn from a large population (n = 1282) of patients. All subjects had been referred for palatoplasty to the same tertiary cleft center between 1978 and 1991. There were 16 matched controls. The listening team included parents of subjects (n = 32) and teachers of age-matched school children (n = 12). Randomized master audiotape recordings of the study group were presented in blinded fashion to both groups of the adult raters, who were inexperienced in the evaluation of patients with speech dysfunction. An experienced panel of three extramural speech pathologists evaluated the same recordings. In all parameters rated, both parents and teachers showed a consistent tendency to give the subject children more negative ratings than the control children. Expert raters were sensitive to differences in resonance and intelligibility in the control and cleft palate groups. Results of this study differ from similar previous research, indicating that naive peer raters (similar-age children) were insensitive to speech differences in the cleft palate and control groups. PMID- 9393462 TI - Vascular lip enlargement: Part I. Hemangiomas--tenets of therapy. AB - Vascular lesions involving the lips pose a difficult problem for both the surgeon and patient. Their removal by surgery may result in greater disfigurement and impairment than the original lesion. When nonsurgical modalities fail, using a well-planned strategy of sequential procedures can provide excellent results. Many hemangioma patients require judicious serial debulking of excess tissue mass, whereas enlargement from port-wine lesions may require direct aggressive surgery. Over a 10-year period, 38 patients underwent surgery for treatment of vascular lip enlargement. In 27 patients, the lip deformities were caused by hemangiomas. The remaining 11 patients had macrocheilia associated with port-wine vascular malformations. This paper specifically addresses hemangiomas of the lips, tenets for their removal, and reduction strategies. Of the 27 patients with hemangiomas involving the lips, 12 had had some form of previous treatment including corticosteroids (4 patients), embolization (3 patients), laser (3 patients), and interferon (2 patients). All 12 of these patients had unsatisfactory results. Specific tenets for the surgical management of these patients are presented. The distribution of the facial hemangiomas was as follows: 15 patients had isolated involvement of the upper lip, 7 lesions involved the lower lip alone and 5 involved both upper and lower lips. Additionally, 10 of these lesions involved the cheek(s), nose, or chin to some degree. Six patients experienced some form of functional impairment before our evaluation including difficulty with eating or drinking, visual obstruction, and psychosocial problems. All operations were performed following several principles established by the senior surgeon (B.M.Z.). By following the tenets presented in this report, he has achieved near-normal lip form, giving the patient marked improvement in appearance and function. PMID- 9393463 TI - Vascular lip enlargement: Part II. Port-wine macrocheilia--tenets of therapy based on normative values. AB - Port-wine (capillary) vascular malformations that enlarge the lips (port-wine macrocheilia) are challenging reconstructive problems which, as a result, often go untreated. The surgical management of these lesions is not straightforward. Scarification by laser to diminish the discoloration has been performed with good results in some cases. However, laser treatment does little to correct three dimensional tissue deformities such as macrocheilia, which must be addressed surgically. We present our experience with the treatment of port-wine macrocheilia in 11 patients over the 10-year period between 1983 and 1994. Basic principles for surgical and nonsurgical treatment of these patients are also discussed. Normative data about lip dimensions are important to surgical planning. We used 40 male and female volunteers, all less than 30 years of age, as a source for measuring normal lip dimensions, thereby creating a normative database. Preestablished points in the labial and perioral region were marked. Measurements were taken and then averaged. This information was used as a guide for surgical excision of large defects in some patients. In addition, in both the lower and the upper lip, if the opposite side is uninvolved, this database could serve as a template for reconstruction of the affected side. Between 1983 and 1994, 11 patients underwent surgery for port-wine macrocheilia. Of the 11 patients, 1 had previous treatment consisting of laser scarification. Four patients had port-wine vascular malformations involving the upper lip alone, four involved the lower lip alone, and three involved both lips. In six patients, other areas of the face and body were also involved. Our experience has led us to perform earlier surgical intervention than has previously been described for these patients. Basic reconstructive surgical principles and planning based on normative data and templates can lead to excellent results. PMID- 9393464 TI - The mental V-Y island advancement flap in functional lower lip reconstruction. AB - Various flap procedures for the reconstruction of lower lip defects have been described to achieve the basic requirements of a functional repair, namely muscle function and sensation. Flaps elevated from the upper lip or the adjacent cheek may provide a solution, but for larger lower lip defects, preservation of function is difficult. In this article, a new functional lower lip reconstruction technique that includes the transfer of a myocutaneous flap based on the mental neurovascular bundle and on the branches of the facial artery is described. The principle of this reconstructive procedure is to advance the tissues from both sides of the chin as myocutaneous flaps upward to the lip defect, reorienting the muscles of the flap for sphincteric function, and preserving the mental nerve for sensation. The depressor anguli oris and remnants of the orbicularis muscle together with their motor nerve branches are included in the V-Y advancement flap. PMID- 9393465 TI - A re-evaluation of hypopharyngeal reconstruction: pedicled flaps versus microvascular free flaps. AB - Reconstruction of hypopharyngeal defects can be accomplished with the use of pedicled myocutaneous flaps or with microvascular free flaps. The authors contrast their results using the trapezoidal paddle pectoralis major myocutaneous flap with the reported results of two published series of free jejunal flaps. The severity of the defects, preprocedure irradiation, and mix of primary and secondary reconstruction were comparable between series. The benefits, complications, and functional results of either technique also seem to be comparable. However, the authors recommend the trapezoidal paddle pectoralis major myocutaneous flap for its ease of performance, rapidity of surgery, and absence of intraperitoneal approach. With the current effort to achieve comparable results with shorter procedures, and with greater conservation of patient and public resources, the pedicled flap should be reconsidered. PMID- 9393466 TI - Spiral CT angiography: an alternative vascular evaluation technique for head and neck microvascular reconstruction. AB - Facial trauma and head and neck oncologic patients are often destined for extensive reconstructive procedures with microvascular free flaps due to ablative injuries or postoperative defects. The integrity and competence of the vasculature in the head and neck recipient site must be imaged and evaluated preoperatively as an essential prerequisite for the success of the reconstructive transfer. In a prospective study of five patients, we compared conventional angiography, the traditional technique, with a new vascular imaging modality- spiral computed tomographic (CT) angiography. One patient suffered from an extensive, ablative facial trauma, and the other four had undergone mandibulectomy as part of their oncologic therapy. In contrast to conventional angiography, spiral CT angiography is a noninvasive imaging technique, which we found to be characterized by much shorter patient examination time, avoidance of selective cannulation with its attendant risks, improved perception of anatomy, and the ability to rotate the reconstructed images in any plane to obtain the best view of any vessel in question. Disadvantages of spiral CT angiography in imaging vessels include the need for relatively large amounts of contrast medium, great dependence on the skill and experience of the operator, and the need for optimizing the timing of the contrast bolus and the scan. PMID- 9393467 TI - A new design of the iliac crest microsurgical free flap without including the "obligatory" muscle cuff. AB - The iliac crest free flap has undergone a gradual evolution to provide more functional and cosmetic oromandibular reconstructions. The soft-tissue cutaneous component has largely resisted refinement and currently constitutes the flap's principal drawback. Conventionally, the cutaneous vessel's soft-tissue encasement and a protective cuff of abdominal muscle are harvested to ensure skin perfusion. These protective measures, however, produce a bulky flap that is tethered to the bone and difficult to inset into complex three-dimensional defects. A series of anatomic and clinical investigations has confirmed that in 30 percent of individuals, the skin island can be elevated on a dominant cutaneous branch from the deep circumflex iliac artery. Harvesting the skin as an axial pattern flap greatly increases its independence from the bone, improving maneuverability. A small collar of abdominal muscle is incised around the pedicle, obviating the need for the customary 2.5-cm protective muscle cuff. Exclusion of the abdominal muscular component reduces the flap's volume, decreases the need for secondary debulking, and reduces the donor site morbidity. PMID- 9393468 TI - Quantitative facial motion analysis after functional free muscle reanimation procedures. AB - The purpose of this study was to evaluate the success of functional free muscle transfer in patients with chronic facial paralysis using a recently developed quantitative method known as the maximum static response assay of facial motion. A retrospective review of a single surgeon series of six patients with longstanding facial paralysis was performed. The maximum static response assay was performed on all patients preoperatively and serially during the postoperative period. Twenty-seven patients (54 sides) with normal facial function were also evaluated and served as controls. The contralateral normal side in those patients with unilateral facial paralysis (n = 4) also served as a control. Movement of the modiolus during smile was recorded in the x axis and y axis. To determine net smile movement, the vector of movement was calculated by means of the Pythagorean theorem. Vectors were then defined mathematically by calculating direction and magnitude. The average direction of the vector during smile for the normal control population was 58.3 degrees (range 32.5 to 83.1 degrees) from the horizontal through the modioli, and the average magnitude was 10.6 mm (range 4.2 to 20.1 mm). The average preoperative direction for the reanimated sides was 176.8 degrees with a range of 83.3 to 225 degrees. Patients with bilateral paralysis (n = 2) were excluded for calculation of the vectors on the normal contralateral side. The average preoperative direction for the normal contralateral side in patients with facial paralysis was 58.3 degrees with a range of 48.2 to 68.4 degrees. Postoperatively, the average direction of the vector during smile for the reanimated sides improved to a value of 77.6 degrees with a range of 45.7 to 113.8 degrees. The average change in direction of the preoperative reanimated side compared with the postoperative reanimated side was significant (p = 0.01). Postoperatively, the average direction of the vector for the contralateral normal sides was 43 degrees with a range of 11 to 57.2 degrees. The change in direction for the contralateral normal side was not significant (p = 0.18). The average magnitude of the reanimated side improved from a non anatomic 2.8 mm preoperatively (range 0.8 to 6.8 mm) to an anatomic 4.9 mm postoperatively (p = 0.02). The contralateral normal side magnitude decreased from 9.4 mm (range 7.3 to 11.6 mm) preoperatively to 5.7 mm (range 3.8 to 7.7 mm) postoperatively (p = 0.006). More specifically, the absolute change in movement on the reanimated side during smile for the x axis and y axis was 2.3 mm (p = 0.05) and 4.0 mm (p = 0.002), respectively. This corresponded to an absolute change in the magnitude of the vector of 4.6 mm in an anatomic direction. On the contralateral side the absolute change in magnitude during smile from preoperative to postoperative for the x axis and y axis decreased by 1.5 mm (p = 0.13) and 5.3 mm (p = 0.05), respectively. This reflected an absolute change in the magnitude of the vector of 5.5 mm. Functional free muscle transfer in patients with chronic facial paralysis resulted in anatomic recovery of motion in the majority of patients in this series. The maximum static response assay can be used to objectively assess the results of facial reanimation. PMID- 9393469 TI - Life span of silicone gel-filled mammary prostheses. AB - The discussion on possible side effects of implanted silicone has resulted in a growing number of patients inquiring whether or not their mammary prostheses are intact and when failure of the prostheses is to be expected. Between November 1988 and May 1995, 182 patients had their silicone mammary prostheses replaced, repositioned, or removed one to three times. Capsular contraction, dislocation, pain paresthesia, and/or suspected rupture were common indications for surgery. To try and be able to provide an indication as to the correlation of implant age and integrity, we recorded the status of all 426 prostheses observed during secondary surgery. In this selected group of patients, approximately 50 percent of the mammary prostheses with an implant age of 7 to 10 years showed gel bleed or rupture. Applying the survival Kaplan-Meier curve, 50 percent of implants may be expected to bleed or be ruptured at the age of 15 years. Rupture was observed more frequently than gel bleed. It seems that there is no chronologic relation between gel bleed and rupture. PMID- 9393470 TI - Textured surface, nonsilicone gel breast implants: four years' clinical outcome. AB - Since the development of smooth silicone breast implants in 1962, more than 1 million women throughout the world have opted for breast augmentation surgery. Although initially successful, smooth implants are prone to develop surrounding scar capsules that may harden and contract, resulting in discomfort, weakening of the shell with rupture, asymmetry, and patient dissatisfaction. This phenomenon has been shown to occur in as many as 70 percent of implanted patients over time. We have reviewed all of our patients and the Medical Device Reporting System for Bioplasty, Inc. (Minneapolis, Minn.) for the history of this device. At 18 months, more textured implants remain soft than the smooth group. After an additional 30 months of follow-up, for a total of 48 months maximum and 18 months minimum, most textured implants still remain soft. Since 1990 we have used AU24K, bio-oncotic hydrogel filling material in molecular impact surface textured implants (MISTI) that is similar to breast tissue in color, radiodensity, and viscosity. Complications have been late leaks, infection, and capsular contracture. Several asymptomatic implants were removed because of anxiety over the FDA controversy. Our experience so far indicates that such a breast implant is a reasonable alternative to the prior art. The longer-term performance of these implants must await the availability of further clinical outcome data. PMID- 9393471 TI - Influence of underfilling on breast implant deflation. AB - The authors report a series of 407 patients with a total of 709 saline breast implants (average follow-up, 7.1 years). In this retrospective series, the overall deflation rate was 6.6 percent (47 of 709). Initial comparison of the deflation rates for smooth (8.8 percent) versus textured (1.8 percent) implants suggested a significant difference between the implant types. However, further analysis of the data revealed that smooth implants had a longer average follow-up period and tended to have lower fill volumes. These data were re-examined using Kaplan-Meier survival analysis plots, which corrected for differences in follow up times, and log rank tests performed to determine significance. Implant type was found to have a non-significant association with rupture rate. In contrast, the percent fill (implant fill volume per minimum recommended fill volume x 100) was significantly associated with the spontaneous ruptures; a mean difference of 13.9 percent (89.2 percent versus 103.1 percent) was found between the series of deflated implants and the nondeflated implants (p < 0.0001). These data suggest that underfilling is a major cause of deflation. PMID- 9393472 TI - The vertically based deep fascia turnover flap of the leg: anatomic studies and clinical applications. AB - Although fasciocutaneous turnover flaps are a simple and fast method for covering soft-tissue defects of the lower leg, many reconstructive surgeons have their doubts about them. They revolve around the lack of criteria for safely designing these random-pattern flaps and around the risk of donor site problems. A vertically based deep fascia turnover flap with a paratibial or parafibular pedicle is presented. Anatomic studies of 36 injected lower limbs showed the deep fascia to be supplied by a mean of 61 vessels. As musculofascial, septofascial, and periosteofascial branches, these contribute to a richly anastomosing vascular network within the deep fascia. Along the deep transverse septum at the medial tibial border, the anterior and posterior peroneal septa, and between the anterior tibial and extensor muscles, the fascia is supplied by segmental vessels in a clearly defined arrangement. Pedicled on these vessels, the deep fascia is a useful candidate tissue for transversely oriented turnover flaps. These are particularly well suited for covering pretibial or prefibular soft-tissue defects. Unlike adipofascial turnover flaps, the transversely oriented deep fascia turnover flap keeps its subcutaneous layer with its intact vascular plexus so that the overlying skin is adequately perfused even in patients with sizable flaps or an extremely thin skin. Clinical experience with the vertically based paratibial or parafibular deep fascia turnover flap in six patients confirmed its usefulness for covering small to medium-sized soft tissue defects of the lower leg. PMID- 9393473 TI - Island adipofascial flap based on distal perforators of the radial artery: an anatomic and clinical investigation. AB - Reconstruction of soft-tissue defects of the hand with exposed tendons, joints, and bone represents a challenge to the plastic surgeon, and such defects necessitate flap coverage to preserve hand function and to protect its vital structures. Reported here is the study of an island adipofascial flap based solely on the distal five to eight septocutaneous perforators of the radial artery and their venae comitantes. Designing the flap in the form of an island with skeletonization of the distal perforators of the radial artery ensures its vascular pattern from these perforators alone with no connection to the ulnar artery perforators or posterior interosseous artery perforators, as is the case with fascial pedicled flaps. Furthermore, designing the flap as an island facilitates the arc of rotation and avoids the pedicle kink when the flap is turned 180 degrees. Preservation of the radial artery, as well as the mild thickness of the flap are further advantages. The drawbacks of such a flap include temporary impaired sensation at the donor site, the obvious scar in the forearm, and loss of hair. Eleven fresh and fixed cadaver upper extremities were dissected to delineate the vascular pattern and to define the arc of rotation of the flap. Also, a clinical approach was conducted on two patients who sustained extension scar contracture with tendon adhesions of the dorsum of the hands, on two patients who sustained first web space contracture, and on two patients who had full-thickness soft-tissue loss over the palm; and finally on two patients who sustained traumatic soft-tissue loss over the dorsum of their hands with exposed tendons and metacarpal bones. PMID- 9393474 TI - The effect of ischemic preconditioning on the recovery of skeletal muscle following tourniquet ischemia. AB - It has been well documented that ischemic preconditioning limits ischemic reperfusion injury in cardiac muscle, but the ability of ischemic preconditioning to limit skeletal muscle injury is less clear. Previous reports have emphasized the beneficial effects of ischemic preconditioning on skeletal muscle structure and capillary perfusion but have not evaluated muscle function. We investigated the morphologic and functional consequences of ischemic preconditioning, followed by a 2-hour period of tourniquet ischemia on muscles in the rat hindlimb. The 2 hour ischemia was imposed without preconditioning, or was preceded by three brief (10 minutes on/10 minutes off) preischemic conditioning intervals. We compared muscle morphology, isometric contractile function, and muscle fatigue properties in predominantly fast-twitch, tibialis anterior muscles 3 (n = 8) and 7 (n = 8) days after ischemia-reperfusion. Two hours of ischemia, followed by reperfusion, results in a 20 percent reduction of muscle mass (p < 0.05) and a 33 percent reduction in tetanic tension (p < 0.05) when compared with controls (n = 8) at 3 days. The same protocol, when preceded by ischemic preconditioning, results in similar decreases in muscle mass and contractile function. Neuromuscular transmission was also impaired in both ischemic groups 7 days after ischemia. Nerve-evoked maximum tetanic tension was 69 percent of the tension produced by direct muscle stimulation in the ischemia group and 65 percent of direct tension in the ischemic preconditioning/ischemia group. In summary, ischemic preconditioning, using the same protocol reported to be effective in limiting infarct size in porcine muscle, had no significant benefit in limiting injury or improving recovery in the ischemic rat tibialis anterior. The value of ischemic preconditioning in reducing imposed ischemic-reperfusion-induced functional deficits in skeletal muscle remains to be demonstrated. PMID- 9393475 TI - Primary critical ischemia time in the hairless mouse ear. AB - The primary critical ischemia time of the hairless mouse ear was determined using a probit analysis. An atraumatic clamp was used across the pedicle base of the ear to induce total ischemia at 0, 2, 4, 6, 7, 7.5, 8.0, 8.5, 9.0, 9.5, and 12 hours of normothermic ischemia. Seventy-seven ears were examined 7 days after the ischemic interval for evidence of necrosis. The median critical ischemia time was determined to be 8.2 hours with 95 percent fiducial limits of 7.74 hours and 8.69 hours. PMID- 9393476 TI - Reverse flow as an option in microvascular recipient anastomoses. AB - This study was designed to investigate the retrograde arterial pressures in the distal ends of the superior thyroid (n = 20) and facial arteries (n = 8). These pressures were compared with mean systemic arterial pressure (n = 20) as well as retrograde pressure in the radial artery (n = 8). The mean retrograde arterial pressure in the radial artery was 50 to 60 percent of normal arterial pressure. Similar pressures were recorded from the distal ends of both the superior thyroid and facial arteries. Because we know that the radial artery can support a skin flap through retrograde or reverse flow (reverse radial forearm flap), it was concluded that both the superior thyroid and facial arteries could also support flaps based on reverse flow. This has proved to be the case clinically. In circumstances where pedicle geometry favors it and in the presence of pulsatile flow from the distal ends of either of these arteries, a retrograde anastomosis is now the practice of the authors in selected cases. PMID- 9393477 TI - Tissue response to suture materials implanted subcutaneously in a rabbit model. AB - We compared the tissue response to a nonabsorbable monofilamented suture made of expanded polytetrafluoroethylene (ePTFE), which has recently been introduced for use in plastic surgery, with the response to 10 other commercially available absorbable sutures and nonabsorbable monofilamented and multifilamented sutures. The sutures were used to secure a patch of ePTFE implanted in the dorsum of adult New Zealand White rabbits. At 30, 60, and 120 days after implantation, the tissue response to the sutures was assessed with respect to the number of foreign-body giant cells present, the thickness of the fibrous capsule that developed, and the general inflammatory response (n = 4 for each suture for each time period). Analysis of variance revealed that specific suture type was significantly associated with foreign-body giant cell count and fibrous capsule thickness. Tevdek had a significantly higher value for mean number of foreign-body giant cells. Silk and Tevdek had significantly thicker fibrous capsules, and ePTFE suture had a significantly thinner capsule. Absorbable sutures and nonabsorbable multifilamented sutures evoked a more extensive tissue response than monofilamented sutures; the differences between nonabsorbable monofilamented and nonabsorbable multifilamented sutures were significant for capsule thickness. In general, suture made of ePTFE produced a minimal tissue response. It should be a good choice for use in facial plastic surgery, in which excellent functional and aesthetic results are critical. PMID- 9393478 TI - The kleeblattschadel anomaly in Apert syndrome: intracranial anatomy, surgical correction, and subsequent cranial vault development. AB - We present a case of Apert syndrome in which intracranial anomalies of the cranial base were localized to the lesser wings of the sphenoid and sphenoid ridge. The lesser wings of the sphenoid were displaced superiorly to follow the fused coronal sutures bilaterally, where they met at a single point on the skull vertex. Careful preoperative study of the intracranial anatomy in the kleeblattschadel anomaly led to a surgical plan for early correction of the anomaly. The present report indicates that an aggressive approach to the correction of the kleeblattschadel anomaly beginning early in infancy can result in normalization of the trilobar skull configuration. Although this approach can correct the kleeblattschadel anomaly, 3.5-year follow-up in this patient with Apert syndrome demonstrates progressive turricephaly despite repeated cranial vault remodeling. Although the trilobar skull configuration can be corrected through early surgical intervention, the long-term correction of progressive turricephaly in patients with Apert syndrome remains an unsolved problem. PMID- 9393479 TI - Chronic orbital hematic cysts: a case for craniofacial correction. AB - Chronic hematic cysts are rare conditions that usually present to the ophthalmic surgeons with displacement of the globe. There is usually no, or minimal, bone involvement. Two patients with unusual presentations of chronic orbital hematic cysts are reported. These cysts resulted in significant expansion and erosion of the bony orbits. The presentation, operative findings, and reconstruction are reported and discussed. PMID- 9393481 TI - Tensor fasciae latae flap: alternative donor as a functioning muscle transplantation. AB - The functioning tensor fasciae late muscle was used for a patient with both a complete defect of the deltoid muscle and a defect of overlying skin. The configuration of the tensor fasciae latae including the length of the muscle belly as well as the muscle fiber arrangement was similar to that of the deltoid. In addition, the spatial relationship between the muscle, donor vessels, and motor nerve fulfilled the requirements for the recipient site of deltoid reconstruction. The transferred muscle successfully replaced the function of the deltoid and provided sufficient strength for elevation of the arm. Simultaneous skin coverage was also satisfactory. The case report here clearly indicates that the tensor fasciae latae muscle is a promising candidate for functioning muscle transplantation, and can also be applied for other disorders. However, several points such as limited motor nerve length should be considered when tensor fasciae latae is used as a functioning muscle. PMID- 9393480 TI - Congenital leukemia cutis: an unusual manifestation of a rare disease. AB - This paper discusses a case of congenital leukemia cutis of lymphoblastic type presenting as a solitary frontonasal tumor. The presentation is unusual when compared with other reported cases of neonatal leukemia and represents the only reported case with comparable presenting features to the authors' knowledge. The differential diagnosis with other frontonasal tumors is discussed. PMID- 9393482 TI - Endoscopic harvesting of the gracilis muscle for reinnervated free-muscle transfer. AB - Reinnervated functioning free-muscle transfer has proven to be invaluable in numerous reconstructive procedures. However, one problem that remains unsolved after transferring the muscle is the presence of a long and cosmetically unacceptable scar at the donor site. This scar has undermined patients' satisfaction with the procedure despite its excellent functional results. In an attempt to resolve this problem, the authors harvested the gracilis muscle endoscopically and now report their technique and results. To create an optical cavity in harvesting the gracilis muscle endoscopically, they devised a lifting apparatus, which is described. Comparative study showed that endoscopic harvesting of the gracilis produced a significantly shorter scar, but took 1.5 times longer than conventional method. PMID- 9393483 TI - Are patients satisfied with results from residents performing aesthetic surgery? AB - An 8-year survey of patient satisfaction in an academic aesthetic surgery clinic at the University of Toronto was carried out by means of a mailed questionnaire. A total of 265 questionnaires were mailed; 131 completed questionnaires (49.4 percent) were returned. Of these, 93.1 percent would recommend this clinic (88.1 percent in the first year of operation and 95.4 percent in the subsequent 7 years), and 92.9 percent would undergo the same procedure again, if required (88.3 percent in the first year and 95.0 percent in the next 7 years). The highest patient satisfaction (10 of 10) was seen in augmentation mammoplasty (average, 9.1); blepharoplasty (average, 9.0); rhytidectomy (average, 7.8), and rhinoplasty (average, 6.9). The results obtained compared favorably with recently published data of more experienced surgeons. PMID- 9393484 TI - SMAS fixation to the facial skeleton: rationale and results. AB - The trend in modern facial rejuvenation surgery is to reposition the ptosis of the superficial soft-tissue mass relative to the facial skeleton. The logical method of supporting the superficial tissue is to reattach it to the underlying skeleton, thereby replicating the function of the retaining ligaments. Five hundred consecutive face lifts involving deep fixation of the submucosal aponeurotic system flap to the periosteum of the zygoma were reviewed for complications and side effects. Expected complications, such as irregularities, dimpling, and palpable sutures, occurred in fewer than 1 percent of cases, confirming the safety of this method. The conclusion from this experience is relevant to all methods of deep-layer facial rejuvenation surgery, including the endoscopic approach. PMID- 9393485 TI - Full face and neck laser skin resurfacing. AB - Since the inception of laser skin resurfacing for the removal of facial rhytides, laser surgeons have avoided laser resurfacing of the neck. The purpose of this paper is to demonstrate that the accuracy and precision of the Ultrapulse carbon dioxide laser allows the laser surgeon to safely laser skin resurface the neck as well as the full face. This series includes 40 patients who have undergone laser skin resurfacing of the neck at the time of full-face laser skin resurfacing. Three subgroups are defined. Thick-skinned patients, thin-skinned patients, and patients who otherwise would have medical contraindications to face and neck lift are included in this study group. Patients between the ages of 40 and 60 who do not have a lot of excessive neck skin or prominent platysma bands are candidates for full face and neck laser skin resurfacing. Successful tightening for thick skinned patients occurs by using 300 mJ at 60 W. a density of 6, and one pass. For thinner-skinned patients, the upper half of the neck is treated with 300 mJ, 60 W, a density of 6, and one pass. The lower half of the neck in these patients is treated with 125 mJ and 20 W, a density of 6, and one pass. In some patients who otherwise have medical contraindications for face and neck lift, the laser may be an indicated procedure because there is minimal bruising, lack of bleeding, minimal edema, minimal to absent use of adrenaline, and nonincisional surgery with a speedy recovery. The Ultrapulse laser delivers high energy with high speed, precision, and control. Therefore, the laser surgeon can successfully laser skin resurface the neck at the time of full face laser skin resurfacing. Immediate tightening of the face and neck, from the photothermal effect, and the neocollagenesis effectively tighten the neck in the properly selected patient. PMID- 9393486 TI - Postoperative care following CO2 laser resurfacing: avoiding pitfalls. AB - Facial skin resurfacing using the carbon dioxide laser has become an increasingly popular procedure. Improvements in carbon dioxide laser technology have made the procedure simpler and more reliable. However, difficulties in the postoperative period can lead to patient morbidity and physician anxiety. Common problems such as prolonged erythema, hyperpigmentation, acne, milia, dermatitis, and infection can be controlled or avoided with proper postoperative care. Less common sequela such as hypertrophic scarring and prolonged healing are often a results of errors committed in the postoperative period. The authors have performed laser resurfacing in almost 2100 patients in the last 4 years. Changes in the postoperative regimen to include no pretreatment, use of semipermeable dressings, antiviral and antibacterial prophylaxis, and early treatment with sunscreens and bleaching agents have made for a smoother recovery with more predictable results. PMID- 9393487 TI - Use of preoperative subcutaneous "wetting solution" and epidural block anesthesia for liposuction in the office-based surgical suite. AB - Uniform saturation of subcutaneous fat using the "wetting solution" formula described by Klein for his "tumescent technique" has been shown to decrease operative blood loss associated with liposuction procedures and to eliminate the requirement for general anesthesia for selected patients. However, we found this infusate provided an inadequate level of anesthesia for many of our patients. We use preoperative infusion of Klein's epinephrine and lidocaine containing wetting solution in our lipoplasty practice only for control of blood loss and postoperative pain. Our anesthetic of choice for liposuction is the epidural block technique, which provides consistent intraoperative comfort for the patient. We report our experience with 85 consecutive lipoplasty patients who underwent liposuction under epidural anesthesia after subcutaneous fat perfusion with Klein's wetting solution. Our epidural block technique uses the rapidly metabolized local anesthetic agent, chloroprocaine, which has the lowest systemic toxicity risk of any local anesthetic agent. Chloroprocaine's anesthetic characteristics are particularly well suited for the outpatient surgery patient with few undesirable side effects. PMID- 9393488 TI - Advances in hair restoration surgery. PMID- 9393489 TI - A simple technique for correction of male nipple hypertrophy: the "sinusoidal" nipple reduction. PMID- 9393491 TI - The art of miscommunication. PMID- 9393490 TI - Complications of the tumescent formula for liposuction. PMID- 9393492 TI - Gore-Tex facial implants. Plastic Surgery Educational Foundation DATA Committee. PMID- 9393493 TI - Vitamin E and wound healing. Plastic Surgery Educational Foundation DATA Committee. PMID- 9393494 TI - External fatty tissue massage (the "endermologie" and "silhouette" procedures). Plastic Surgery Educational Foundation DATA Committee. PMID- 9393495 TI - Computer imaging/surgical simulation. Plastic Surgery Educational Foundation DATA Committee. PMID- 9393499 TI - Fat trapper. PMID- 9393496 TI - Advances in assessing outcome of surgical repair of cleft lip and cleft palate. PMID- 9393497 TI - Physiopathology of the dynamic muscular sphincter of the pharynx. PMID- 9393498 TI - Influence to the development of cleft lip, palate, or both by fertilized ovum transfer in A/J strain mice. PMID- 9393500 TI - Task force on ultrasound-assisted lipoplasty. PMID- 9393501 TI - Correction of the elevated nasal ala after Millard's repair in unilateral cleft lip. PMID- 9393502 TI - Subcutaneous infiltration in suction-assisted lipoplasty. PMID- 9393503 TI - A new transconjunctival retractor. PMID- 9393504 TI - Saphenous vein injection in the rat: a simple and time efficient technique. PMID- 9393505 TI - Temporalis muscle flap revisited on its centennial: advantages, newer uses, and disadvantages. PMID- 9393506 TI - Gluteal fasciocutaneous V-Y advancement flap. PMID- 9393507 TI - Function of the infrahyoid muscle flap. PMID- 9393508 TI - Device for easy inflation of expanders used in breast reconstruction. PMID- 9393509 TI - On call. PMID- 9393510 TI - Method for middle vault reconstruction in primary rhinoplasty: upper lateral cartilage bending. PMID- 9393511 TI - High-resolution imaging of the musculoskeletal system. PMID- 9393512 TI - Human immunodeficiency virus infection and hepatitis: biosafety in radiology. AB - Radiologists frequently perform invasive diagnostic and therapeutic procedures involving needles and/or vascular access, and often they do so in darkened rooms. Therefore, they are at risk of exposure to blood-borne pathogens. The risk of HIV infection with a single sharp injury is low (0.3%), and on average 99.7% of exposures will not result in infection. However, this seroconversion rate is increased when a high volume of blood or a high concentration of virus is inoculated, and it is decreased by 79% when postexposure prophylaxis is used. An estimated 800,000 needle-stick injuries and other injuries from sharp objects to health care workers occur annually in the United States (25). Approximately 16,000 of these involve HIV-contaminated blood, and even more are contaminated with HBV or HCV (46). Needle-stick injury therefore poses the single greatest risk to health care workers regarding occupational transmission of HIV. Because most patients in the radiology department have an unknown HIV or hepatitis serostatus, all patients should be regarded as potentially infectious, and precautions should be universal. In fact, the 1991 OSHA ruling made compliance with the CDC Universal Precautions Guidelines the enforceable national standard. Real-time oral communication among all members of the radiology team and scrupulous attention to safe technique are absolutely essential. Radiologists are not in agreement regarding the use of precautions against injury with a sharp object and splashing (47-50). Many have adapted some of their habits to conform well to the CDC and OSHA guidelines regarding universal precautions, but some remain skeptical regarding the risk of exposure to themselves. Consequently, in some areas resistance to the above recommendations persists. However, the data to date provide a compelling argument for protection against occupational exposure to blood either by percutaneous sharp injury or splashing on mucous membranes or interrupted skin. A number of resources were made available in early 1997 for easy access to the most current data regarding occupational transmission of HIV or hepatitis. For instance, the CDC has a World Wide Web site (http://www.cdc.gov) and a facsimile information service through the Hospital Infections Program directory (telephone 404-332-4565). Also, the National AIDS Clearinghouse can be reached by telephone (800-458-5231), as can the HIV/AIDS Treatment Information Service (800-448-0440). The postexposure prophylaxis protocol used at the University of California, San Francisco, can be reviewed by visiting its World Wide Web site at http://epi-center.ucsf.edu. And up-to-date information is available to both Veterans Administration and other health care staff worldwide by J. Michael Howe, MSLS, of the AIDS Information Center, a service of the VA HIV/AIDS National Training Program, located at the Veterans Administration Medical Center, San Francisco, University of California, San Francisco (hivinfo@itsa.ucsf.edu). PMID- 9393513 TI - Imaging squamous cell carcinomas of the upper aerodigestive tract: what clinicians need to know. PMID- 9393514 TI - Valediction: sweet sorrow. PMID- 9393515 TI - Duplex sonography: can it be used to evaluate carotid artery stents? PMID- 9393516 TI - Improving radiology research methods: what is being asked and who is being studied? PMID- 9393517 TI - Traumatic thoracic aortic aneurysm: treatment with endovascular stent-grafts. AB - PURPOSE: To demonstrate the feasibility and safety of endovascular stent-graft placement for treatment of traumatic aortic aneurysm. MATERIALS AND METHODS: Ten patients with traumatic aortic aneurysm were treated with endovascular stent grafts. Three patients had an acute traumatic aneurysm; seven had a chronic aneurysm. Stent-grafts were constructed from modified Z-stents covered with woven polyester or expanded polytetrafluoroethylene graft material and were deployed through a 20-24-F delivery sheath in an exposed artery located remotely from the lesion. RESULTS: Stent-graft placement and thrombosis of the aneurysmal sac were successful in all patients. Major complications were encountered in three patients after endovascular treatment. One patient had a peri-graft leak; complete thrombosis of the aneurysmal sac was achieved after coil embolization of the leak. Transposition of the left subclavian artery was necessary to relieve left arm ischemia in another patient. In the third patient, stent placement in the left main stem bronchus was needed to relieve left lung atelectasis. All patients were alive and without complications during the follow-up period (mean, 15 months). CONCLUSION: Transluminal placement of endovascular stent-grafts is a technically feasible method for treatment of traumatic thoracic aortic aneurysm and may be an effective alternative to open-chest surgery. PMID- 9393518 TI - Percutaneous biliary drainage: clinical trial of analgesia with interpleural block. AB - PURPOSE: To determine the analgesic efficacy and safety of interpleural block for percutaneous biliary drainage. MATERIALS AND METHODS: In this double-blind study, 34 age- and sex-matched patients who were to undergo percutaneous biliary drainage because of malignant biliary obstruction were randomly assigned to the true-block group (30 mL 0.5% bupivacaine block) or placebo-block group; all had access to a patient-controlled analgesia (fentanyl) pump. Self medication, pain reports, blood pressure, heart rate, and oxygen saturation were monitored during and until 8 hours after drainage. The McGill Pain Questionnaire was administered 1 hour after biliary drainage. RESULTS: Patients in the placebo group self administered statistically significantly more fentanyl than did patients in the true-block group (P = .008). Peak pain scores (10-point scale) and McGill Pain Questionnaire scores were statistically significantly higher for the placebo group patients (P = .017 and P = .001, respectively). There were no differences between groups in terms of blood pressure, heart rate, and oxygen saturation. Two patients had pneumothorax caused by the interpleural block. CONCLUSION: Interpleural block was effective in decreasing pain and opioid requirements during and after percutaneous biliary drainage and did not compromise the cardiopulmonary status of the patient. However, the rate of pneumothorax was higher than previously reported. PMID- 9393519 TI - Radiologically guided placement of pull-type gastrostomy tubes. AB - PURPOSE: To evaluate percutaneous placement of pull-type gastrostomy tubes that has traditionally necessitated endoscopic guidance. MATERIALS AND METHODS: From September 1995 through March 1997, 63 pull-type gastrostomy tubes were placed in 64 patients. Retrograde catheterization of the esophagus was performed through the stomach. Then the gastrostomy tube was pulled through from the mouth into the stomach. RESULTS: Gastrostomy tube placement was successful in 63 (98%) of 64 patients in 65 attempts. One procedure was stopped when the patient reported chest pain after gastric insufflation, and a second placement attempt was initially unsuccessful. Major complications occurred in three (5%) patients: exit site infection necessitating tube removal (n = 2) and prolonged bleeding necessitating transfusion (n = 1). Minor complications occurred in six (9%) patients: failure of placement (n = 2), exit site infection (n = 1), leakage around the tube (n = 1), tube migration (n = 1), and inadvertent tube removal (n = 1). There were no cases of peritonitis, tract disruption, or gastrostomy related death. CONCLUSION: Percutaneous placement of a pull-type gastrostomy tube was performed with a minimum risk of tract disruption and peritonitis. The tube was safely and effectively placed by radiologists. PMID- 9393520 TI - Rapid thrombectomy with a hydrodynamic catheter: results from a prospective, multicenter trial. AB - PURPOSE: To evaluate efficacy and safety of a hydrodynamic rheolytic thrombectomy device for rapid percutaneous treatment of acute thromboembolic occlusions of native lower-extremity arteries and bypass grafts. MATERIALS AND METHODS: In 50 patients, thrombectomy was performed with the rheolytic catheter at four centers. Patients had acute occlusions of native lower-extremity arteries (n = 39) or acute thrombosis of lower-limb bypass grafts (n = 11). Estimated occlusion age was 5 days +/- 5. Mean thrombus length was 15 cm +/- 11. Clinical success was measured on a scale of -3 (deterioration) to +3 (improvement) with established criteria. RESULTS: With the thrombectomy catheter, the majority of thrombus material was removed and antegrade blood flow was reestablished in 45 (90%) patients. Technical success (residual luminal narrowing < 50%) was 52% with use of the device alone. Adjunctive therapy was performed in 45 patients. Clinical improvement after intervention was +3 in 25 (50%) patients, +2 in 10 (20%), +1 in six (12%), and 0 (no improvement) in nine (18%). Clinically unimportant complications related to use of the device were one (2%) distal embolization and two (4%) dissections. Laboratory analysis revealed hemolysis without clinical sequelae. Primary patency rates were 76% after 30 days, 74% after 3 months, and 69% after 1 year. CONCLUSION: The hydrodynamic catheter appears to be safe and effective for rapid thrombectomy. PMID- 9393521 TI - PTFE-encapsulated endovascular stent-graft for transjugular intrahepatic portosystemic shunts: experimental evaluation. AB - PURPOSE: To evaluate the safety and efficacy of a stent-graft designed for a transjugular intrahepatic portosystemic shunt (TIPS), assess angiographic and hepatic biologic responses to polytetrafluoroethylene (PTFE)-encapsulated stents, and compare with a bare stent. MATERIALS AND METHODS: Twelve TIPS (eight with flexible PTFE-encapsulated balloon-expandable stent-grafts and four control TIPS with bare Wallstents) were created in 12 pigs. Shunt venography was performed at 1-month intervals and necropsy of graft-containing animals at 1, 2, 3, 4, and 5 months. Control animals were sacrificed at 6 weeks. Detailed histopathologic analyses were performed. RESULTS: The stent-grafts were readily deployed in all cases. Seven of eight graft TIPS remained fully patent during the follow-up period without luminal encroachment. Typical myofibrolasts proliferated on the abluminal surface of the graft, without extension into the lumen. No inflammatory reaction was present. Cellular overgrowth from the hepatic vein occluded the end of one graft at 3 months, partly related to rapid axial growth of that animal. The endoluminal surface of this shunt was otherwise patent. At 4-6 weeks, one control TIPS was occluded and the other three showed 45%-85% stenoses. No bile staining was seen in any case. CONCLUSION: This PTFE-encapsulated stent-graft is biocompatible and safe to place. It markedly improves TIPS patency, providing almost uninterrupted, unimpeded patency in this model. PMID- 9393522 TI - Renal artery stenosis: evaluation with breath-hold, three-dimensional, dynamic, gadolinium-enhanced versus three-dimensional, phase-contrast MR angiography. AB - PURPOSE: To compare breath-hold, three-dimensional, gadolinium-enhanced magnetic resonance (MR) angiography with three-dimensional, phase-contrast MR angiography in the evaluation of renal artery stenosis. MATERIALS AND METHODS: Fifty-five consecutive adult patients with clinical suspicion of renovascular disease were prospectively examined with three-dimensional, phase-contrast MR angiography and breath-hold, three-dimensional MR angiography with injection of a standard dose of gadopentetate dimeglumine to evaluate the number of renal arteries and the presence and degree of stenosis. The standard of reference was intraarterial digital subtraction angiography. RESULTS: Gadolinium-enhanced MR angiography depicted all 105 main renal arteries, whereas phase-contrast MR angiography depicted 104. The number of accessory renal arteries depicted was significantly higher with gadolinium-enhanced (17 of 18) than with phase-contrast (11 of 18) studies (P = .04). Both techniques depicted 27 of the 29 stenoses (sensitivity, 93%; P > .05). Sensitivities, specificities, and accuracies in the diagnosis of hemodynamically significant stenosis (> 50% narrowing) were, respectively, 94%, 96%, and 96% for phase-contrast and 100%, 97%, and 98% for gadolinium-enhanced MR angiography (P > .05). CONCLUSION: Gadolinium-enhanced MR angiography is superior to phase-contrast MR angiography in accessory renal artery depiction. No statistically significant difference in the assessment of stenosis has been found between the two techniques. PMID- 9393523 TI - Coronary artery calcification in women with syndrome X: usefulness of double helical CT for detection. AB - PURPOSE: To determine the usefulness of double-helical computed tomography (CT) for detection of diseased coronary arteries in women with anginal pain, positive exercise stress test results, and angiographically normal coronary arteries (syndrome X). MATERIALS AND METHODS: Double-helical CT of the coronary arteries was performed in 81 consecutive women who were referred for coronary angiography for evaluation of chest pain. Patients were classified into three groups according to stress test and angiographic results: normal (normal exercise test results and angiographically normal coronary arteries), syndrome X (abnormal exercise test results and angiographically normal coronary arteries), and coronary artery disease (at least one diseased vessel seen at angiography). RESULTS: The prevalence of coronary calcification in the syndrome X group was 63% (10 of 16 patients) compared with 96% (45 of 47 patients) in the coronary artery disease group (P = .002) and 22% (four of 18 patients) in the normal group (P = .02). The lowest total coronary calcification score and logarithmic transformed data were found in the normal group (2.9 +/- 0.7), statistically significantly higher values were found in the syndrome X group (4.3 +/- 1.5), and the highest values were found in the coronary artery disease group (5.1 +/- 2.0; for trend, P = .03). CONCLUSION: Double-helical CT may be useful in detection of atherosclerosis in women with syndrome X who demonstrate normal coronary arteries at angiography. PMID- 9393524 TI - Visualization of colorectal polyps with spiral CT colography: evaluation of processing parameters with perspective volume rendering. AB - PURPOSE: To evaluate two key processing steps for detection of colon polyps with spiral computed tomographic (CT) colography with perspective volume rendering (PVR): image reconstruction and opacity assignment of the attenuation data. MATERIALS AND METHODS: Spiral CT was performed in 10 patients with known polyps confirmed at colonoscopy, and detailed quantitative analyses were performed of data obtained in four. First, anatomic fidelity of three-dimensional (3D) images generated from two-dimensional (2D) source images with equal voxel dimensions (87%-90% overlap) was compared with 3D images generated from 2D source images with unequal voxel dimensions (0%-80% overlap). Next, the relative dimensions of colorectal polyps to adjacent structures were evaluated for various opacity threshold settings. Then, step and sigmoidal opacity functions were compared with respect to image smoothness and edge sharpness. RESULTS: PVR images generated after interpolation of image data reconstructed with at least 60% overlap were equivalent in image quality to PVR images generated from source images with equal voxel dimensions. Relative polyp-to-haustral fold dimensions demonstrated substantial distortions with opacity thresholds below -700 HU. The 3D PVR images generated with the sigmoidal opacity function were significantly smoother than those generated with the step opacity function (paired t test, P < .02), with small differences noted in edge sharpness. CONCLUSION: Use of highly overlapping source images (87%-90%) was not necessary to generate 3D PVR images of colorectal polyps. Image artifacts were suppressed with use of an appropriate opacity threshold and a sigmoidal opacity function without substantial loss in edge sharpness. PMID- 9393525 TI - Hypervascular liver metastases: do unenhanced and hepatic arterial phase CT images affect tumor detection? AB - PURPOSE: To evaluate the relative roles of unenhanced and hepatic arterial phase (HAP) computed tomographic (CT) imaging in the detection of hypervascular liver metastases. MATERIALS AND METHODS: Eighty-four patients with biopsy-proved liver metastases from hypervascular primary tumors other than hepatocellular carcinoma underwent unenhanced and HAP and portal venous phase (PVP) helical CT studies. Three blinded radiologists evaluated each series of images separately for the number, size, and enhancement characteristics of lesions. Sixty-nine patients had follow-up imaging proof of tumor burden. RESULTS: The three readers detected 381 402 lesions on the PVP images and 397-416 lesions on the unenhanced images. Unenhanced images allowed detection of 72%-80% of the lesions seen on PVP images. They detected 94-137 additional lesions on unenhanced but not PVP images. On the HAP images, 375-395 lesions were identified. HAP images allowed detection of 81% 90% of the lesions seen on PVP images. Forty-five to 78 additional lesions were detected on HAP but not on PVP images. In the 69-patient subset, maximal detection of tumor foci occurred in 94% of patients with unenhanced plus PVP images and in 78% with HAP plus PVP images. Unenhanced plus PVP images allowed detection of 96% of the 322 tumors in the subset population. CONCLUSION: Unenhanced plus PVP CT images allow detection of statistically significantly more hypervascular liver metastases than do HAP plus PVP images or imaging only in the PVP. PMID- 9393526 TI - Colorectal cancer: diagnostic potential of CT measurements of hepatic perfusion and implications for contrast enhancement protocols. AB - PURPOSE: To assess changes in hepatic perfusion in patients with colorectal cancer with computed tomography (CT), diagnostic potential of CT perfusion measurements, and implications for design of contrast enhancement protocols. MATERIALS AND METHODS: In 27 patients with colorectal cancer, arterial and portal perfusion were calculated from temporal changes in attenuation after intravenous administration of contrast material. RESULTS: Arterial perfusion greater than 0.25 mL/min/mL was seen in nine (82%) of the 11 patients with overt metastases versus six (38%) of the 16 patients with no overt metastases (P < .05). Portal perfusion of 0.25 mL/min/mL or less was found in five (46%) of the patients with overt metastases versus three (19%) of the patients with no overt metastases. Follow-up imaging showed progressive metastatic disease in three patients, all of whom had decreased portal perfusion. CONCLUSION: Increased arterial perfusion appears to be an indicator of liver metastases, whereas reduced portal perfusion may indicate progressive disease. Contrast enhancement protocols that are based on experience with normal livers may not be optimal for patients with metastases. PMID- 9393528 TI - Small (1.5 cm or less) liver metastases: US-guided biopsy. AB - PURPOSE: To determine the techniques used for and the success of ultrasound (US) guided biopsy of hepatic metastases 1.5 cm in diameter or smaller. MATERIALS AND METHODS: A computer search of radiology reports identified 29 patients who underwent US-guided biopsy of 30 hepatic masses 1.5 cm in diameter or smaller suspected to be metastases. All 30 lesions were sampled for biopsy with the free hand technique. Parameters assessed were lesion size and location, needle size, transducer type, number of passes made, cytologic or histologic analysis, and final histologic diagnosis. Biopsies were considered successful if a positive histologic diagnosis of metastasis was made. RESULTS: The mean lesion diameter was 1.3 cm (range, 0.9-1.5 cm). Lesion depth was 3-9 cm (mean, 5 cm). Twenty biopsies were performed with a 22-gauge aspirating needle and analyzed cytologically. An average of 2.7 passes were made per lesion. Phased-array sector transducers were used in 23 lesions. In 28 (93%) lesions and 28 (96%) patients, an adequate specimen was obtained to establish the histologic diagnosis of metastatic disease. CONCLUSION: US appears to be an effective guidance technique for biopsy of small liver metastases. PMID- 9393527 TI - Hepatocellular carcinoma: evaluation with dynamic and static gadobenate dimeglumine-enhanced MR imaging and histopathologic correlation. AB - PURPOSE: To analyze the potential of gadobenate dimeglumine-enhanced magnetic resonance (MR) imaging for the characterization and diagnosis of hepatocellular carcinoma (HCC) by using static and dynamic sequences. MATERIALS AND METHODS: Twenty-eight patients with histopathologically proved HCC were evaluated with T1- and T2-weighted spin-echo and static and dynamic gradient-echo sequences before, during, and after intravenous administration of 0.1 mmol/kg gadobenate dimeglumine (0.5 mol/L). RESULTS: During the perfusion phase of the dynamic sequence, all 16 nodular well-differentiated HCC lesions showed a rapid increase in signal intensity 10-30 seconds after injection followed by a progressive decrease in signal intensity. The nine poorly differentiated HCC lesions showed no rapid increase in signal intensity. All eight large (> 3 cm), nodular, well differentiated HCC lesions showed a hypointense rim before injection and both hypo- and hyperintense rims (double-ring sign) immediately after injection, compared with normal liver parenchyma. About 55 seconds after injection, substantial single-rim enhancement was detected in 21 of the 28 HCC lesions. CONCLUSION: Dynamic gadobenate dimeglumine-enhanced MR imaging allows improved characterization of HCC lesions, which show rapid increase in signal intensity during the early, arterial phase in well-differentiated HCC lesions and a double ring sign in large well-differentiated nodular HCC lesions. PMID- 9393529 TI - Gastrointestinal submucosal tumors: evaluation with endoscopic US. AB - PURPOSE: To describe the endoscopic ultrasound (US) features of benign versus malignant submucosal tumors throughout the gastrointestinal tract. MATERIALS AND METHODS: One hundred nine patients aged 24-81 years suspected to have submucosal tumors (11 esophageal, 41 stomach, 24 duodenal, and 33 colorectal tumors) at barium studies or endoscopy underwent endoscopic US. The layer of origin, internal echo pattern, and lesion margin were analyzed by means of consensus and independent interpretation by three radiologists. RESULTS: Endoscopic US findings revealed several distinct patterns among various submucosal tumors. Sixteen (94%) of the 17 homogeneous lesions with histopathologic findings of malignancy were hypoechoic, although 29 (43%) of the 68 homogeneous lesions with histopathologic findings of benignity were similarly hypoechoic. Homogeneous lesions that were anechoic, of intermediate echogenicity, or hyperechoic were almost exclusively benign (39 [98%] of 40). In contrast, 23 (96%) of the 24 malignant lesions were heterogeneous (n = 7) or homogeneously hypoechoic (n = 16). The sizes of benign and malignant lesions were significantly different (P < .05). There was no significant difference in the echo pattern (i.e., homogeneous versus heterogeneous), but there was a significant difference in the proportion of hypoechoic versus nonhypoechoic lesions (anechoic, hyperechoic, or of intermediate echogenicity; P < .001). CONCLUSION: The differential diagnosis of gastrointestinal submucosal tumors is assisted with endoscopic US. PMID- 9393530 TI - Cystic dystrophy of the duodenal wall: radiologic findings. AB - PURPOSE: To determine the radiologic characteristics of cystic dystrophy of the duodenal wall. MATERIALS AND METHODS: Ten patients with cystic dystrophy of the duodenal wall and chronic pancreatitis underwent ultrasonography (US) (n = 10), computed tomography (CT) (n = 10), endoscopic US (n = 5), and endoscopic retrograde cholangiopancreatography (ERCP) (n = 9). Cystic dystrophy of the duodenal wall was classified as either cystic or solid. The imaging findings were retrospectively analyzed and compared with findings at pancreatoduodenectomy (n = 10). RESULTS: The more frequent cystic type (n = 7) of cystic dystrophy of the duodenal wall was characterized by the presence of easily recognizable cystic lesions (diameter, more than 1 cm), located within the thickened wall of the second portion of the duodenum. The solid type (n = 3) of cystic dystrophy of the duodenal wall demonstrated fibrous thickening of the duodenal wall within which small cysts (diameter, less than 1 cm) were present. The intraduodenal cysts were usually elongated or bilobate with a thick wall. The thickening of the duodenal wall appeared as a solid layer between the duodenal lumen and the pancreas, hypoechoic at US, isoattenuating at unenhanced CT, and hypoattenuating in the early phase (after initiation of infusion of contrast material) and isoattenuating in the late phase (after completion of infusion) at contrast material-enhanced CT. Findings at retrospective analysis of CT and endoscopic US images were characteristic. CONCLUSION: Imaging modalities, notably CT and endoscopic US, helped establish the diagnosis of cystic dystrophy of the duodenal wall. PMID- 9393531 TI - Carotid artery stents: early and intermediate follow-up with Doppler US. AB - PURPOSE: To determine whether ultrasound (US) is a sensitive follow-up method after placement of a carotid artery stent for the detection of significant stenosis, occlusion, and other complications at early and intermediate follow-up. MATERIALS AND METHODS: Doppler US examinations were performed after stent placement in 170 carotid arteries in 119 patients with angiographic correlation. Prospective diagnostic US criteria for stenosis were peak-systolic velocity greater than 1.25 m/sec, internal carotid artery (ICA) to common carotid artery (CCA) peak-systolic velocity ratio of greater than or equal to 3:1, and intrastent doubling of peak-systolic velocity. Retrospective criteria for stenosis were also applied: peak-systolic velocity greater than 1.7 m/sec, ICA end-diastolic velocity greater than 0.4 m/sec, ICA/CCA peak-systolic velocity ratio greater than 2.0, and ICA/CCA end-diastolic velocity ratio greater than 2.4. RESULTS: Eighty-seven immediate and 83 intermediate (average, 7.3 months) follow-up US examinations were performed. Two stent occlusions were detected. One or more prospective US criteria were abnormal in 26 arteries with a stent. One or more retrospective criteria were positive in 47 arteries. Angiography showed corresponding findings, with only one significant stenosis (63%) in the ICA stents. Moderate collapse of a CCA stent was depicted at US. CONCLUSION: Only one significant recurrent stenosis was detected, and no significant stenoses were missed at US. US successfully depicted carotid artery stent occlusion and a moderate stent collapse. Sensitivity in the detection of intrastent stenosis is promising. Further study to refine US criteria in a study with longer term follow up is needed owing to the lack of significant recurrent stenosis in the intermediate follow-up group. PMID- 9393532 TI - Normalizing fractional moving blood volume estimates with power Doppler US: defining a stable intravascular point with the cumulative power distribution function. AB - PURPOSE: To normalize the power Doppler ultrasound (US) signal to the expected signal from 100% blood in the calculation of a fractional moving blood volume estimate. MATERIALS AND METHODS: To locate the signal from flowing blood with a consistent backscatter coefficient, the authors estimated the knee of the cumulative Doppler power distribution function. They used a flow-tube phantom to test the use of this knee to locate a radial position that would fall into a region of high shear stress and minimal rouleaux formation. They also studied how well the method normalized fractional moving blood volume estimates of the right renal cortex in a volunteer when simulating different body habitus and in a group of six healthy volunteers to estimate variability. RESULTS: Over five flow velocities and over undersaturated to severely oversaturated receiver gains, the calculated flow-tube area was a mean 89% +/- 7 (+/- standard deviation) of a standard. In humans, the technique normalized the fractional moving blood volume estimates over an 8-dB receiver gain variation; the mean +/- standard deviation of fractional moving blood volume estimates for the six volunteers was 37.6% +/- 3.6. CONCLUSION: Vascularity estimates with power Doppler US are feasible with a normalization scheme based on the cumulative Doppler power distribution function. PMID- 9393533 TI - Renal cancer: preoperative evaluation with dual-phase three-dimensional MR angiography. AB - PURPOSE: To evaluate the use of dual-phase three-dimensional magnetic resonance (MR) angiography in the preoperative staging of renal cancer. MATERIALS AND METHODS: MR angiography was performed in 18 patients before performance of partial (n = 7), radical (n = 10), or laparoscopic (n = 1) nephrectomy to treat renal cancer. Dynamic, three-dimensional MR angiograms were obtained with gadoteridol enhancement, breath holding, and a three-dimensional spoiled gradient echo sequence. Images were obtained at 15-second intervals to achieve opacification of arteries and veins. Source, maximum intensity projection, and multiplanar reconstruction images were evaluated. Imaging results were compared with surgical findings. RESULTS: Renal arterial phase MR angiograms depicted 30 of 31 (97%) surgically confirmed renal arteries, with one false-positive result (an artery that arose from an early-branching single main renal artery, interpreted as a separate accessory artery). Renal venous phase MR angiograms depicted all seven instances of renal vein involvement, including extension to the inferior vena cava in two patients. Collateral vessels around the tumor, including prominent gonadal veins in three patients, were demonstrated. Additional findings included adenopathy and adrenal and pulmonary metastases. CONCLUSION: Dual-phase MR angiography of the kidney may be a useful technique in depicting renal vessels before nephrectomy. PMID- 9393534 TI - Placenta accreta: evaluation with color Doppler US, power Doppler US, and MR imaging. AB - PURPOSE: To determine the value of transabdominal ultrasound (US), transvaginal US, color Doppler US, power Doppler US, and magnetic resonance (MR) imaging in the diagnosis of placenta accreta. MATERIALS AND METHODS: Nineteen patients in the third trimester of pregnancy who were at risk for placenta accreta underwent color Doppler and power Doppler US; 18 patients also underwent MR imaging. Images were interpreted prospectively for signs of accreta by two reviewers. The reviewers' confidence in their diagnosis was graded on a five-point scale. RESULTS: Outcomes at delivery were as follows: normal placenta (n = 11), hysterectomy owing to uncontrollable bleeding (n = 1), and placenta accreta (n = 7). Five cases of lower-uterine-segment placenta accreta were diagnosed with a high level of confidence with vaginal and power Doppler US. In one patient with a posterior placenta who had previously undergone myomectomy, MR imaging enabled the diagnosis of placenta accreta, which was not well depicted at US. CONCLUSION: In patients with a history of uterine scars, vaginal US with power Doppler US performed well in the evaluation of lower-uterine-segment placenta accreta. MR imaging depicts posterior placenta accreta. PMID- 9393535 TI - Medial border of the perirenal space: CT and anatomic correlation. AB - PURPOSE: To explore the mode of spread of disease between the perirenal space and the perivascular central retroperitoneum and to determine the anatomy along the medial border of the perirenal space. MATERIALS AND METHODS: Anatomic dissection, injection of latex, and observation of cross sections of the abdomen were performed in nine cadavers. Attention was paid to the juncture of the central prevertebral, perivascular, and extraperitoneal regions, and the perirenal space. Anatomic findings were correlated with observations made at computed tomography (CT) in 82 patients with retroperitoneal hemorrhage (n = 24), inflammation (n = 37), and neoplasia (n = 21) involving the perirenal spaces or the central retroperitoneum. RESULTS: Along most of the length of each kidney, no apparent fascia separates the perirenal space from the central retroperitoneum. At this location, septa between fat lobules form a fenestrated multitier barrier. These septa were imperceptible on CT scans obtained in healthy individuals. After injection of latex in cadavers, this potential barrier was seen. In the clinical study, spread of disease was allowed in only 38 (30%) of 128 instances of potential spread. Spread was facilitated along the renal vessels and the interlobular septa. CONCLUSION: Beyond the kidneys, the renal fascia is closed, forming a cone superiorly and an inverted cone inferiorly. A network of interlobular septa acted as a barrier or pathway to the free spread of disease from the perirenal space to the central retroperitoneum or from the central retroperitoneum to the perirenal space. PMID- 9393536 TI - Balloon dacryocystoplasty: indications and contraindications. AB - PURPOSE: To define the indications and contraindications for balloon dacryocystoplasty. MATERIALS AND METHODS: Eighty-five patients with severe epiphora due to partial (n = 47) or complete (n = 38) obstruction of the nasolacrimal duct (NLD) were treated with balloon dacryocystoplasty (DCP). Steerable micro-guide wires with flexible tips were used. Success rates of DCP were evaluated clinically and dacryocystographically during the acute phase and at 6- and 12-month follow-up. Failures and recurrences were correlated with clinical and dacryocystographic indications for treatment. RESULTS: Recanalization was successful in 35 (92%) of 38 patients with isolated focal stenoses (n = 20) or short-distance occlusions (n = 18) of the NLD. Among all 85 patients, recanalization was successful in 25 patients (66%) with complete and 37 patients (79%) with partial obstructions. In the absence of the main predictors for recurrent obstructions (ie, active inflammation, filling defects due to calculi, long-distance occlusions, and posttraumatic lesions), 12-month patency rates were 89% (17 of 19 focal stenoses) and 94% (15 of 16 focal occlusions). Otherwise, reobstruction rate was 46% (12 of 26 cases). CONCLUSION: Balloon dacryocystoplasty is successful only in select cases. To achieve results comparable to those of operative treatment, the indication should be limited to patients with circumscribed focal stenoses or occlusions of the NLD. Active dacryocystitis, dacryocystolithiasis, and posttraumatic lesions are the main contraindications. PMID- 9393537 TI - Obstructed nasolacrimal duct system in epiphora: long-term results of dacryocystoplasty by means of balloon dilation. AB - PURPOSE: To evaluate the long-term results of balloon dacryocystoplasty in the treatment of epiphora due to obstruction of the nasolacrimal ducts. MATERIALS AND METHODS: One hundred eyes in 84 patients with complete or incomplete obstruction of the lacrimal sac and duct were selected for dacryocystoplasty. A catheter with a balloon diameter of 3 mm was used. Follow-up was 5-48 months. No stents were placed. A Kaplan-Meier analysis was used to evaluate patency. RESULTS: The long term primary patency rate was 70% +/- 7 (+/- standard error). Repeat dacryocystoplasty was successful in 10 of the 11 cases with initial failure or reobstruction during follow-up, which yielded a long-term secondary patency rate of 81% +/- 7. There was no association between the length of the obstruction or the duration of symptoms before dacryocystoplasty and the initial and long-term success. Initial and long-term success was statistically significantly higher in dacryocystoplasty for an incomplete obstruction rather than for a complete obstruction. CONCLUSION: The long-term results of dacryocystoplasty, followed if necessary by repeat dacryocystoplasty, are good. Dacryocystoplasty is a safe and simple procedure and could become the treatment of choice for epiphora due to obstruction of the nasolacrimal ducts. Dacryocystorhinostomy is indicated when dacryocystoplasty or repeat dacryocystoplasty fails or when dacryocystoplasty is contraindicated (e.g., in anatomic malformations in the lacrimal duct or bony canal). PMID- 9393538 TI - Brain capillary telangiectasia: MR imaging appearance and clinicohistopathologic findings. AB - PURPOSE: To demonstrate the clinical and magnetic resonance (MR) imaging findings of brain capillary telangiectasia and compare them with postmortem specimens. MATERIALS AND METHODS: MR images obtained in and clinical histories of 18 adult patients with a presumed diagnosis of capillary telangiectasia examined within 3 years were retrospectively reviewed. All patients had undergone MR imaging with conventional T1- and T2-weighted spin-echo sequences and gadolinium-enhanced T1 weighted and susceptibility-sensitive gradient-echo (GRE) sequences. No biopsies had been performed. Fourteen patients had undergone clinical and MR imaging follow-up (median, 11 months). Postmortem tissues from three cases of histopathologically confirmed capillary telangiectasia were imaged. RESULTS: All lesions were small, homogeneously enhancing, and hypo- to isointense on T1 weighted images and iso- to slightly hyperintense on proton-density- and T2 weighted images. None was hypointense on proton-density- or T2-weighted images. All lesions showed marked GRE signal loss. None had changed at follow-up. Two patients had multiple classic cerebral cavernous angiomas. The three specimens showed no abnormal susceptibility and contained no hemosiderin at tissue analysis. CONCLUSION: Capillary telangiectasia has mild contrast material enhancement but is otherwise undetectable on conventional MR images. It lacks the "hemosiderin rim" of cavernous angioma and demonstrates increased susceptibility only on GRE images, likely owing to blood oxygen-level-dependent contrast. GRE is essential in diagnosing brain capillary telangiectasia, which could otherwise be misdiagnosed as neoplasia, subacute infarction, or demyelination. PMID- 9393539 TI - Intracranial tumor in children: MR imaging findings within 24 hours of craniotomy. AB - PURPOSE: To evaluate whether very early magnetic resonance (MR) imaging enables distinction of residual tumor from benign postoperative change in children. MATERIALS AND METHODS: Forty-six postoperative MR examinations were performed in 43 children with intracranial tumors within 24 hours of the completion of surgery during a 2-year period. These examinations were categorized according to whether residual tumor could be definitely identified or excluded, or whether the diagnosis was uncertain. RESULTS: Contrast enhancement occurred in 33 of 46 MR examinations performed within 24 hours of surgery. In 18 instances, this was associated with obvious residual tumor. In 15 patients, only small amounts of linear or patchy enhancement were seen. Of these, seven patients (46%) were disease-free for an average of 4.5 years. Assessment for postoperative enhancement was hampered in seven patients because of the presence of methemoglobin in the tumor bed. Contrast enhancement was not observed in two patients before surgery. CONCLUSION: Surgically induced, MR-detectable contrast enhancement and extracellular methemoglobin formation occurs within 24 hours of the completion of intracranial surgery. This can interfere with the detection of small amounts of residual tumor. PMID- 9393540 TI - Fetal cerebral ventriculomegaly: misidentification of the true medial boundary of the ventricle at US. AB - PURPOSE: To investigate the implications of mistaking the medial surface of the cerebral hemisphere for the medial wall of the lateral ventricle at antenatal ultrasonography (US) and to identify US clues that might help avoid this interpretive error. MATERIALS AND METHODS: In 50 second- and third-trimester fetuses, a directed attempt was made to demonstrate the medial surface of the cerebral hemisphere and the medial wall of the lateral ventricle on images that depicted the lateral wall of the ventricle. In each fetus, measurements of the diameter of the false ventricular atrium were compared with the true diameter of the lateral ventricle to assess the potential magnitude of error. RESULTS: The average diameter measured with the medial surface of the cerebral hemisphere was 10.7 mm, compared with the true mean ventricular diameter of 6.2 mm. This value was greater than or equal to 10 mm (the generally accepted upper limit of normal for the ventricular diameter) in all 15 third-trimester fetuses and in 16 (46%) of 35 second-trimester fetuses. The parietal occipital fissure was demonstrated along the medial surface of the cerebral hemisphere in 36 (72%) of 50 fetuses, and the medial surface of the cerebral hemisphere could be traced posteriorly around the occipital lobe in 45 (90%). CONCLUSION: When ventriculomegaly is suspected, the examiner should make a direct attempt to find the medial wall of the ventricle and distinguish it from the medial boundary of the cerebral hemisphere. Correct identification of the anatomic interfaces is facilitated by demonstrating that the cerebral interface contains the parietal occipital fissure and can be traced posteriorly around the occipital lobe. PMID- 9393541 TI - Pneumonia in children: decreased parenchymal contrast enhancement--CT sign of intense illness and impending cavitary necrosis. AB - PURPOSE: To determine if computed tomographic (CT) findings of decreased contrast material enhancement are predictive of more intense illness and of the development of cavitary necrosis in children with pneumonia. MATERIALS AND METHODS: Contrast-enhanced CT scans in 44 children with pneumonia who did not respond appropriately to therapy were compared with precontrast CT scans to evaluate enhancement of consolidated lung parenchyma. Enhancement was correlated with admission to the intensive care unit, length of hospital stay, cavitary necrosis in the lung at follow-up CT, and frequency of lung resection. RESULTS: Parenchymal enhancement was decreased in 21 children; pneumonia was enhanced in the other 23 children. Decreased enhancement was associated with increased admission to intensive care (14 of 21 [67%] vs five of 23 [22%] children; P = .0026), increased length of hospital stay (15 vs 10 days; P = .0615), increased frequency of cavitary necrosis at follow-up CT (seven of seven [100%] vs none of three children; P = .0086), and increased frequency of resection (two of 21 [10%] vs none of 23 children). At histopathologic examination, diffuse cavitary necrosis was present in resected lobes in two patients. CONCLUSION: Decreased parenchymal enhancement at CT is a predictor of more intense illness and may herald the development of cavitary necrosis in children with pneumonia. PMID- 9393542 TI - Vesicoureteral reflux in older children: concordance of US and voiding cystourethrographic findings. AB - PURPOSE: To determine if a negative renal sonogram is reliably predictive of the absence of vesicoureteral reflux at voiding cystourethrography (VCUG) in children aged 5 years or older. MATERIALS AND METHODS: Imaging studies in 70 children aged 5 years or older who underwent renal ultrasound (US) and VCUG on the same day were reviewed. These children had initially undergone evaluation because of a urinary tract infection. RESULTS: Five of 70 children had abnormal sonograms; two (40%) of the five had reflux at VCUG. One had mild pelvicalyceal dilatation, and one had a small kidney. The other three (without reflux) had a pelvic kidney, a calyceal diverticulum, or a renal stone. Of 65 children with a negative sonogram, 19 (29%) had reflux at VCUG; 46 (71%) did not. Altogether, of the 70 children, 21 had reflux, 19 (90%) of whom had no sonographic abnormality. CONCLUSION: Children with abnormal screening renal sonograms often have vesicoureteral reflux, but a normal sonogram does not reliably exclude the condition even in children aged 5 years or older. Therefore, VCUG must be performed even in older children, regardless of US findings, if clinical decisions are influenced by documentation of the presence of VUR. PMID- 9393543 TI - Detection of malignant and benign breast lesions with an automated US system: results in 120 cases. AB - PURPOSE: To evaluate clinically an automated ultrasound (US) system for detecting benign and malignant breast lesions. MATERIALS AND METHODS: A prototype automated US system was used to examine 119 patients: 38 patients with 39 proved malignant breast lesions (7-50 mm), 41 patients with 41 proved benign breast lesions (8-40 mm), and 40 patients without breast lesions. The device yields a three dimensional set of B-mode scans and reconstructed US images comparable to mammograms. All patients had undergone mammography. Four radiologists who had not performed the examinations independently assessed the mammograms and US images to detect benign and malignant breast lesions. RESULTS: Each of the four readers did not recognize one to three detectable malignant lesions on mammograms, one to two detectable malignant lesions on US images, two to four detectable benign lesions on mammograms, and five to seven detectable benign lesions on US images. All readers identified the 39 cancers with at least one of the modalities. The 40 cases without lesions were diagnosed correctly more frequently on the US images by three readers and on the mammograms by one reader. CONCLUSION: Depiction of breast lesions at automated US is reproducible. Automated US is complementary to mammography. PMID- 9393544 TI - Metastatic breast carcinoma in axillary lymph nodes: in vitro US detection. AB - PURPOSE: To establish the ultrasonographic (US) characteristics of benign versus metastatic lymph nodes. MATERIALS AND METHODS: One hundred fifty-eight axillary lymph nodes in 40 patients (age range, 31-73 years) surgically treated for breast cancer have been studied in vitro with a 7.5-MHz US probe in a water bath. The long-to-short axis ratio and the hilar and cortical characteristics were evaluated; the US findings were correlated with the histopathologic findings. To estimate the long-to-short axis ratio, all lymph nodes were measured. RESULTS: Of the 158 lymph nodes, 45 showed histopathologic evidence of metastasis; 38 of the 45 revealed US signs of malignancy. The signs that caused malignancy to be suspected were a long-to-short axis ratio of less than 1.5, absence of a hilus, and disruption of the cortical zone. The most specific sign for the diagnosis of metastasis was absence of the hilus. The increase in the long-to-short axis ratio was the finding that caused the most false-negative interpretations. Signs of malignancy were more accurate in lymph nodes 10 mm or larger than they were in lymph nodes smaller than 10 mm. CONCLUSION: Findings of in vitro US studies of axillary adenopathy provide the basis for the evaluation of lymph node metastasis in vivo before surgery, especially in those lymph nodes 10 mm or larger. PMID- 9393545 TI - Dynamic echo-planar imaging of the breast: experience in diagnosing breast carcinoma and correlation with tumor angiogenesis. AB - PURPOSE: To correlate quantitative echo-planar magnetic resonance (MR) imaging measures of gadopentetate dimeglumine tumor uptake with histologic diagnoses and microvessel density (MVD) and to compare dynamic echo-planar imaging of breast lesions with conventional dynamic MR imaging techniques. MATERIALS AND METHODS: The study group comprised 63 patients (aged 13-70 years) with 71 breast lesions who underwent conventional and echo-planar MR imaging. The T1 values, change in gadopentetate dimeglumine concentration, and extraction-flow products were calculated with the echo-planar imaging data and were correlated with histologic findings and MVD estimates. Extraction-flow product data normalized to pectoral muscle gadopentetate dimeglumine concentration in invasive cancers was also correlated with MVD. RESULTS: On average, cancer T1 values were shorter than benign values, but there was substantial overlap between the two groups. Cancers had higher extraction-flow products than benign lesions (P < .001). Sensitivity, specificity, positive predictive value, and negative predictive value were 83%, 79%, 67%, and 90%, respectively. Receiver operating characteristic analysis showed improved performance with extraction-flow products than with percentages of signal intensity change. Among the invasive cancers, there was no significant correlation between extraction-flow product and MVD. CONCLUSION: The T1 value remains important in more precise quantitative estimation of gadopentetate dimeglumine uptake in breast tumors, which helps improve the specificity of dynamic imaging. Tumor MVD affects the contrast medium enhancement of breast lesions, but other factors contribute. PMID- 9393546 TI - Functional anatomy of the thoracic outlet: evaluation with spiral CT. AB - PURPOSE: To determine the anatomic characteristics of the thoracic outlet before and after dynamically induced modifications. MATERIALS AND METHODS: Fifty-two volunteers (24 women; 28 men; mean age, 42 years) with no clinical or radiographic indications of thoracic outlet syndrome underwent spiral computed tomography (CT) of the apexes at full inspiration with the arms alongside the body and then with the dominant arm in hyperabduction, with a contralateral rotation of the head. RESULTS: After elevation of the dominant arm, (a) no statistically significant difference was found in median value of the costosubclavian and costoclavicular distances; (b) the median distance between the posterior border of the smaller pectoral muscle and the anterosuperior chest wall was 12 mm in all subjects; (c) the subclavian artery in 18 (75%) women and in 20 (71%) men and/or the subclavian vein in three (12%) women and in three (11%) men were identified in the costoclavicular space. The median angles of rotation, retraction, and upward displacement of the clavicle were 22 degrees, 32 degrees, and 7 degrees, respectively, in women and 25 degrees, 31 degrees, and 11 degrees, respectively, in men. CONCLUSION: Spiral CT is expected to be useful for determining the complex pathophysiologic processes that underlie thoracic outlet syndrome. PMID- 9393547 TI - The breast: in-plane x-ray protection during diagnostic thoracic CT--shielding with bismuth radioprotective garments. AB - PURPOSE: To evaluate the ability of thin overlying bismuth radioprotective shielding to reduce the x-ray dose to radiosensitive superficial organs during diagnostic computed tomography (CT). MATERIALS AND METHODS: A variety of patient and phantom studies were performed with four thicknesses of bismuth radioprotective latex over the breast. Dose savings were determined with thermoluminescent dosimeters. A prototype and then a final manufactured radioprotective brassiere was constructed and tested for radiation dose savings to the breast during diagnostic chest CT. Preliminary studies were also performed to evaluate shielding of the thyroid, orbit, and testes. RESULTS: The use of bismuth radioprotective latex saved an average 57% of the radiation dose to the breast from thoracic CT, decreasing the radiation level from an average 2.2 rad (0.022 Gy) to 1.0 rad (0.010 Gy) (P < .001). Preliminary tests of shielding other superficial radiosensitive organs frequently included at diagnostic CT (eyes, thyroid gland, and testes) were performed with the same thickness of overlying bismuth radioprotective latex, with similar results. Radiation to the thyroid gland was reduced by 60% (from 0.0573 to 0.0229 Gy) and radiation to the eye and testes was reduced by 40% (from 0.0256 to 0.0154 Gy) and 51% (from 0.0463 to 0.0229 Gy), respectively. CONCLUSION: The use of in-plane overlying bismuth radioprotective latex manufactured into form-fitting garments did not affect the diagnostic CT image but reduced the amount of radiation to radiosensitive superficial structures. PMID- 9393549 TI - Scaphopisocapitate alignment: criterion to establish a neutral lateral view of the wrist. AB - PURPOSE: To determine whether a "true" neutral lateral view of the wrist is necessary for accurate capitolunate angle measurement, to compare two standards of diagnostic adequacy for neutral lateral wrist views (distal radioulnar overlap [RUO] and scaphopisocapitate [SPC] relationship), and to confirm positional reproducibility and measurement precision of the SPC criterion. MATERIALS AND METHODS: Capitolunate angles were measured on neutral lateral and supine pisotriquetral views of 10 normal wrists. Two hundred neutral lateral wrist views were classified by each standard (RUO and SPC) as excellent, acceptable, or unacceptable. In two subgroups, capitolunate angles were measured on the lateral views to determine SPC practicality and sensitivity. RESULTS: Compared with neutral lateral positioning, supinated off-lateral views showed an apparent increase in lunate dorsiflexion of up to 30 degrees. Diagnostically unacceptable, excellent, and acceptable pronosupination was present on 118, 22, and 60 of 200 views by using the RUO criterion and on 40, 79, and 81 of 200 views by using the SPC criterion, respectively. The capitolunate angle did not show a significant difference between each of the two subgroups (P > .05). CONCLUSION: A true neutral lateral view of the wrist is necessary for accurate measurement of the capitolunate angle on the basis of a comparison with off-lateral views. SPC relationship provides a diagnostically reproducible standard for a neutral lateral wrist view and should reduce the need for repeat lateral radiographs. PMID- 9393548 TI - Accuracy of bedside chest hard-copy screen-film versus hard- and soft-copy computed radiographs in a medical intensive care unit: receiver operating characteristic analysis. AB - PURPOSE: To compare the clinical diagnostic accuracy of hard-copy readings of screen-film bedside chest radiographs and both hard- and soft-copy readings of bedside chest computed radiographs obtained in a medical intensive care unit. MATERIALS AND METHODS: Two samples of 95 cases were assembled from chest images obtained in 541 patients with either screen-film radiography or computed radiography. The cases were stratified according to the clinical problem for which the examination was ordered; the corresponding diagnosis was verified by a panel of two or three radiologists. Four radiologists blindly read the hard-copy images obtained with screen-film or computed radiography. Six months later, the radiologists read the computed radiographs by using an 8-bit, 1,684 x 2,048-pixel display. The data were analyzed by using multireader-multicase receiver operating characteristic (ROC) analysis of variance. RESULTS: No statistically significant differences in the area under the ROC curve were found between any of the methods. CONCLUSION: The results provide some justification for using bedside chest computed radiography and for reading soft-copy images from a high-quality display. PMID- 9393550 TI - Soft-tissue sarcoma involving bone or neurovascular structures: MR imaging prognostic factors. AB - PURPOSE: To determine if magnetic resonance (MR) imaging findings of soft-tissue sarcoma involving bone or neurovascular structures allow prediction of local recurrence, distant metastasis, or disease-specific survival. MATERIALS AND METHODS: Preoperative MR images of 46 patients with soft-tissue sarcoma were reviewed for tumor involving bone or major vessels or nerves. MR imaging findings were correlated with local recurrence, distant metastasis, and disease-specific survival after surgery and chemotherapy and/or radiation therapy. Primary tumors were predominantly in the lower extremity (n = 35 [76%]), deep (n = 44 [96%]), and high-grade (n = 35 [76%]). RESULTS: On MR images, bone invasion occurred in 12 patients (26%), major-vessel encasement in five patients (11%), and major nerve encasement in seven patients (15%). In patients with (n = 12) and those without (n = 34) bone invasion, frequencies of disease-related death (in nine [75%] and 12 [35%] patients, respectively) were significantly different (P = .02); frequencies of local recurrence or distant metastasis were not significantly different. In patients with and those without major-vessel or major nerve encasement, there were no significant differences between frequencies of local recurrence, distant metastasis, or disease-specific survival. CONCLUSION: In soft-tissue sarcoma, bone invasion on MR images was predictive of decreased disease-specific survival. MR imaging findings of bone or neurovascular involvement otherwise appear to be more important for tailoring surgery than for predicting local recurrence, distant metastasis, or survival. PMID- 9393551 TI - Bell's palsy and herpes simplex virus. AB - Bell's palsy, which is defined as idiopathic peripheral facial paralysis of sudden onset, accounts for > 50% of all cases of facial paralysis. Different theories on the etiology of Bell's palsy have been proposed and investigated. Various clinical studies have suggested an etiological link between Bell's palsy and herpes simplex virus (HSV). In addition, animal experiments have shown the ability of HSV to induce facial paralysis. In our opinion, the possible link between Bell's palsy and HSV can only be explored properly by studying the human facial nerve, and especially the geniculate ganglion itself. Different groups have tried to detect hypothetically reactivated and hypothetically latent HSV in the facial nerves of Bell's palsy patients and control patients, respectively. The isolation of infectious HSV from facial nerve tissue by conventional cell culture methods appeared to be very difficult, also when Bell's palsy patients were tested. Instead, modern molecular methods, such as in situ hybridization and the polymerase chain reaction (PCR) could easily detect HSV DNA in geniculate ganglia. The detection of HSV-specific latency-associated transcripts in the ganglia of control patients provided further evidence for the hypothetically latent state of HSV in the geniculate ganglia in these patients. Recent PCR experiments performed by a Japanese group strongly suggest that the area adjacent to the geniculate ganglia does not usually contain any HSV at all, except in patients with Bell's palsy. This well-controlled study provides conclusive evidence that reactivation of HSV genomes from the geniculate ganglia is the most important cause of Bell's palsy. Consequently, it has been suggested that "Bell's palsy" be renamed as "herpetic facial paralysis". PMID- 9393552 TI - Normal interleukin-12 production in individuals with antibodies to Helicobacter pylori. AB - It is increasingly recognized that the inability of the immune system to clear H. pylori infection is caused by an inadequate immune response and is associated with chronic gastric inflammation. To further investigate the cellular immune response to H. pylori, we studied PBMC from 31 H. pylori antibody-negative and 16 H. pylori antibody-positive individuals for H. pylori-induced DNA synthesis, secretion of the Th1-type cytokine IFN-gamma and secretion of IL-12, a cytokine produced by bacteria-stimulated monocyte/macrophages and a potent inducer of antibacterial immune responses and Th1-type T cells. All experiments were performed using Y. enterocolitica 03 as control. Our results demonstrate that DNA synthesis, IFN-gamma production and IL-12 production induced by H. pylori or Y. enterocolitica 03 did not differ significantly between H. pylori antibody positive and H. pylori antibody-negative individuals. However, in the H. pylori positive group there was a tendency, although not statistically significant, to produce less IFN-gamma in response to H. pylori but not Y. enterocolitica. These results demonstrate that monocyte/macrophages from H. pylori-positive individuals secrete normal amounts of IL-12 upon bacterial challenge and suggest that the decreased production of IFN-gamma in H. pylori-positive individuals observed in previous studies is selective for H. pylori and not caused by decreased IL-12 secretion. PMID- 9393553 TI - Hemagglutination ability and adherence to the Buffalo green monkey kidney cell line of uropathogenic Escherichia coli. AB - The hemagglutination ability and adherence capacity to the Buffalo green monkey (BGM) kidney cell line of 160 wild-type strains of Escherichia coli isolated from bacteriuric patients were investigated. It was found that P-fimbriated E. coli strains adhered significantly better to BGM cells than did strains in which P fimbriae were not detected, which is in accordance with the capacity of P fimbriated strains to cause unobstructive pyelonephritis and with receptor distribution for P-fimbriae in the urinary tract. The strains which exhibited other adhesions, alone or simultaneously, showed reduced adherence to BGM cells, while non-agglutinating strains, mostly isolated from urine of patients with asymptomatic bacteriuria, did not adhere at all or adhered poorly to the utilized cell line. The BGM cells served as a good experimental model for investigation of uropathogenic E. coli adherence; because these cells originate from the upper urinary tract, they are viable and not coated with Tamm-Horsfall protein. PMID- 9393554 TI - Chronic Pseudomonas aeruginosa lung infection is more severe in Th2 responding BALB/c mice compared to Th1 responding C3H/HeN mice. AB - The chronic Pseudomonas aeruginosa lung infection in cystic fibrosis (CF) is characterized by a pronounced antibody response and microcolonies surrounded by numerous polymorphonuclear neutrophils (PMN). Poor prognosis is correlated with a high antibody response to P. aeruginosa antigens. An animal model of this infection was established in two strains of mice: C3H/HeN and BALB/c, generally known as Th1 and Th2 responders, respectively, which were challenged with alginate-embedded P. aeruginosa. Mortality was significantly lower in C3H/HeN compared to BALB/c mice (p < 0.025). P. aeruginosa was cleared more efficiently in C3H/HeN mice and significantly more C3H/HeN mice showed normal lung histopathology (p < 0.02), and we found significantly fewer microabscesses in C3H/HeN mice than in BALB/c mice (p < 0.005). In supernatants from P. aeruginosa antigen and concanavalin A-stimulated spleen cells from the two strains of mice, the interferon-(IFN-) gamma levels were higher, whereas IL-4 levels were lower in C3H/HeN mice than in BALB/c mice. The implications of these findings for CF patients with chronic P. aeruginosa lung infection are discussed. PMID- 9393555 TI - Improved ELISA for determination of anti-diphtheria and/or anti-tetanus antitoxin antibodies in sera. AB - Double-antigen ELISAs for detection and quantification of anti-tetanus or anti diphtheria antibodies in serum have been developed. The assays showed good correlations with established toxin neutralizing assays and were functionally specific for IgG antibodies. The double-antigen set-up allows specific antibodies to bind to antigen-coated microtitre wells with one arm and the free arm to bind to biotin-labelled antigen. The amount of antibodies able to bind labelled antigen was assessed by adding enzyme-conjugated streptavidin and colour substrate followed by measurement of the colour using an ELISA reader. The double antigen principle makes it possible to compare samples of different species on the same plate, permitting the direct use of existing international references of animal or human origin. The double-antigen ELISAs showed a detection limit of 0.00002 IU/ml for both antibodies and were suitable for quantifying antibodies in blood samples collected on filter paper as well as in serum. The assays required no special equipment compared to traditional ELISA. PMID- 9393556 TI - Antibiotic susceptibility of blood culture isolates of Enterobacteriaceae from six Norwegian hospitals 1991-1992. AB - From August 1991 to February 1992, each of the six largest hospitals throughout Norway collected 84 to 107 consecutive blood culture isolates of Enterobacteriaceae, altogether 571 isolates. The distribution of various species and genera at the different hospitals was uniform; Escherichia coli being most prevalent (57-67%), followed by Klebsiella spp. (12-18%) and Proteus mirabilis (7 11%). Twenty-one and 4% of E. coli isolates were resistant to ampicillin and cefuroxime, respectively, and 11% of Klebsiella isolates were cefuroxime resistant. Five Enterobacter isolates and one Citrobacter isolate were resistant to ceftazidime, and one Salmonella isolate was resistant to imipenem. All isolates were susceptible to ciprofloxacin and tobramycin. These results were compared with the antibiotic consumption in each hospital region. Although hospitals in the regions with the highest consumption of ampicillin tended to have a higher percentage of isolates resistant to this agent, no significant differences were found. There was no significant difference between hospitals regarding prevalence of cefuroxime-resistant isolates. PMID- 9393557 TI - Mice overexpressing human lecithin: cholesterol acyltransferase are not protected against diet-induced atherosclerosis. AB - Lecithin: cholesterol acyltransferase (LCAT) (EC 2.3.1.43) is generally assumed to participate in reverse cholesterol transport, i.e., cholesterol transport from peripheral tissues to the liver. LCAT is secreted by the liver and transported in plasma mostly associated with high density lipoprotein. It catalyzes the esterification of cholesterol, mainly high density lipoprotein cholesterol, and produces cholesteryl ester and lysolecithin. Transgenic mice overexpression human LCAT on a C57BL/6 background have elevated high density lipoprotein cholesterol and markedly reduced low and very low density lipoprotein cholesterol and triglyceride levels in plasma, suggesting that such mice may be less susceptible to diet-induced atherosclerosis than isogenic nontransgenic controls. To determine if the apparent anti-atherogenic lipoprotein profile of the LCAT transgenics reduced their susceptibility to atherogenesis, the atherosclerotic lesions developing in transgenic LCAT mice and controls when fed an atherogenic diet were compared by histology and morphometry. Histological examination of the aortas from mice fed a high fat diet for 12, 17 and 22 weeks revealed that the aortic lesions were no smaller or less developed in the transgenic LCAT mice than in the C57BL/6 controls. After 17 weeks there were significantly more "fatty streaks" in the transgenic mice than in the controls. Thus, overexpression of human LCAT in transgenic mice, in spite of their very favourable blood lipoprotein and lipid profile, does not protect against development of atherosclerosis. PMID- 9393558 TI - Electron microscopic study of milk sediments. Qualitative and quantitative observations. AB - Sediment of milk from ovine mammary glands infected experimentally with coagulase negative staphylococci was examined by transmission electron microscopy. The proportions, of the various particles and cells present in milk and of macrophages and neutrophils involved in phagocytosis were determined 6 h, 18 h and 49 days after infection. All cell types together predominated over cytoplasmic bodies in approximately 54% of normal milk samples. The proportion of macrophages in normal milk was higher than that of neutrophils in approximately 69% of samples examined. After infection and in the early phase of inflammation, the proportion changed in favour of neutrophils. In both the early and late phases of infection, greater numbers of macrophages (3-27.2%) than neutrophils (0 2.7%) contained phagocytized cocci. Moreover, greater numbers of cocci were observed in macrophages than in neutrophils (mean values 8.1 vs 1.2), indicating the importance of the macrophage in the maintenance of ovine subclinical mastitis caused by coagulase negative staphylococci. PMID- 9393559 TI - The antifungal effect of lactoferrin and lysozyme on Candida krusei and Candida albicans. AB - Lactoferrin and lysozyme (muramidase) are non-immune defence factors present in various exocrine secretions, including saliva. Previous studies have shown that both proteins, either singly or in combination, are bactericidal in nature and their combined activity is synergistic. As little is known of their interactions with Candida species, 20 oral isolates of C. krusei and 5 isolates of C. albicans were studied for their susceptibility to human apo-lactoferrin and lysozyme, either singly or in combination, using an in vitro assay system. The two species exhibited significant interspecies differences in susceptibility to lactoferrin (p < 0.05), but not for lysozyme; C. krusei being more sensitive to lactoferrin (c 1.4 times) than C. albicans. Both species revealed significant intraspecies differences in their susceptibility to lysozyme (p < 0.05), but not for lactoferrin. No synergistic antifungal activity of the two proteins on either Candida species was noted. The results imply that the variable expression of the fungicidal activity of lactoferrin and lysozyme on Candida species may modulate the oral carriage of yeasts in a complex manner. PMID- 9393560 TI - Antibodies against human papillomavirus type 6 capsids are elevated in men with previous condylomas. AB - Serum samples from 47 men with current condylomas, 32 men with a history of condylomas and from 205 men with no history of genital wart disease, who were attending sexually transmitted disease (STD) clinics at two different hospitals in Stockholm, were analyzed for the presence of immunoglobulin G (IgG) and A (IgA) antibodies to capsids of human papillomavirus types 6 and 11. IgG to HPV type 6 was found among 35% of patients with a history of condylomas compared to 10% of controls (p = 0.0003), but only among 27% of patients with current condylomas. Antibodies to HPV 6 and to HPV 11 showed a very limited correlation, suggesting that the antibodies are HPV-type restricted. The results strengthen conclusions from a previous serological study indicating that IgG antibodies against HPV 6 develop late during condylomatous disease and mostly reflect previous exposure to the virus. PMID- 9393561 TI - Optimization of a battery using nine immunocytochemical variables for distinguishing between epithelial mesothelioma and adenocarcinoma. AB - A battery of immunocytochemical analyses, previously established to distinguish between malignant mesothelioma and metastatic adenocarcinoma, was extended by analysing the same cases with three other commercially available antibodies. Altogether, 11 antibodies were studied in mesotheliomas diagnosed by other means, using 14 different immunocytochemical parameters. Logistic regression analysis indicated that the following parameters were of importance for this diagnostic problem: vimentin reactivity in epithelial cells (1), cytokeratin (CAM 5.2) reactivity in spindle-shaped (fibrous) cells (2), cell membrane-associated reactivity of EMA (3), HBME-1 (4) and thrombomodulin (5), and absence of reactivity to CEA (6), CD15 (7), BerEp4 (8) and Sialyl-TN (9). The analysis gave an algorithm with which a specific diagnosis of mesothelioma could be made in 80% of the cases-i.e., some improvement compared to the 55% sensitivity using the previous battery. PMID- 9393563 TI - CD73 (ecto-5'-nucleotidase) on blood mononuclear cells. Regulation of ecto-5' nucleotidase activity and antigenic heterogeneity of CD73 on blood mononuclear cells from healthy donors and from patients with immunodeficiency. PMID- 9393562 TI - The pituitary-gonadal function in postmenopausal women with epithelial ovarian tumors. PMID- 9393564 TI - The need for a multidisciplinary approach in the treatment of advanced colorectal cancer: a critical review from a medical oncologist and surgeon. AB - Over the last 10 years important advances have been made in the treatment of patients with advanced colorectal cancer, particularly with surgery either alone or in combination with radiotherapy. Furthermore, despite early scepticism, several chemotherapy studies have now reported significant clinical benefits with 5-FU-based regimens and promising results have also been reported with newer agents such as raltitrexed and irinotecan. Taken together these advances now enable a significant proportion of patients to undergo treatment which will improve their quality of life, prolong survival and even result in cure in certain cases. Patients with advanced colorectal cancer can only benefit from these important advances, however, if a truly multidisciplinary approach to patient care is adopted which requires integration of the roles of the surgeon, medical oncologist and radiotherapist. PMID- 9393565 TI - Lymphatic mapping and selective lymphadenectomy for melanoma: not yet standard therapy. PMID- 9393566 TI - A one-year audit of 255 operable breast cancers. AB - The publications 'Guidelines for Surgeons in the Management of Symptomatic Breast Disease' and 'QA Guidelines for Surgeons in Breast Cancer Screening' by the BASO Breast Specialty Group set standards for audit which were aimed at good, achievable practice. Data from 251 patients with operable breast cancers, under the age of 70 years and treated in 1994 at Nottingham City Hospital's Professorial Unit of Surgery, were audited according to 12 of the quality objectives and outcomes measures specified. The questions addressed included: waiting time for first appointment; number of attendances in diagnostic clinic; time for results to be given; pre-operative diagnosis; waiting time for surgery; localization biopsy reports; and number of therapeutic operations. Six outcome measures achieved the targets, four were close and required minimal action for correction, while two were not achieved. As a result of this audit corrective action has been taken with regard to the latter two quality objectives. PMID- 9393567 TI - Percutaneous self-expandable metallic stents and malignant biliary strictures. AB - Thirty-five patients with malignant obstructive jaundice received palliative treatment using percutaneous self-expandable metallic stents. Cholangiocarcinoma was the most frequent cause of the biliary obstruction. In more than 50% of cases, the stricture was located in the hilum. Adequate biliary drainage was achieved in 97% of cases. Median survival was 182 days, and 11% of patients died within 30 days. Early complications occurred in 31% of patients, and 25% of patients showed recurrent jaundice after an average of 180 days. Percutaneous self-expandable metallic stents are an efficient means of palliatively treating malignant biliary strictures, particularly high biliary obstructions. PMID- 9393568 TI - Palliative and adjuvant regional chemotherapy in pancreatic cancer. AB - To improve the dismal prognosis of patients (pts) with pancreatic cancer we treated 32 patients with non-resectable (UICC III, 17 pts; UICC IV, 15 pts--group 1) and 20 patients with resected (UICC I, 1 pt; UICC II, 3 pts; UICC III, 16 pts- group 2) pancreatic cancer with palliative (group I) and adjuvant post-operative (group II) coeliac axis intra-arterial cyclic infusions (CAI). CAI consisted of mitoxantrone 10 mg/m2 on day 1, folinic acid 170 mg/m2 and 5-FU 600 mg/m2 during days 2-4, and cis-platinum 60 mg/m2 on day 5 for up to 11 (group I) or six (group II) cycles. In a total of 211 cycles toxicities at the level of WHO III occurred in 0-6% and of WHO IV in 0%. The median survival times, compared with institutional historical controls (treated vs controls), were 12 vs 4.8 months in UICC III (P < 0.006) and 4 vs 2.9 months in UICC IV (P < 0.05) group I pts, and 21 vs 9.3 months in group II (P < 0.0003). Hepatic disease progression appeared to be suppressed with CAI, which also appears to be effective for palliative and adjuvant treatment in non-resectable and resected pancreatic cancer. PMID- 9393569 TI - Expression of p53 in recurrent nodal metastasis from nasopharyngeal carcinoma (NPC). AB - This study reports the incidence of p53 expression in 40 patients with recurrent nodal metastasis from nasopharyngeal carcinoma (NPC) and its prognostic value in this group of patients. Immunohistochemical staining using monoclonal antibody specific for human p53 protein was performed on the tumour-bearing nodes from 40 patients. The results were divided into four grades (I, negative; II, < 10% of cells positive; III, 10-50% of cells positive; and IV, > 50% of cells positive). The staining scores were correlated with histological tumour types, subsequent recurrence and survival. All patients had undergone neck irradiation. Lymph node specimens from six patients (15%) showed positive staining of nuclear p53 protein. The distribution among the different grades was: three (7.5%) for II, two (5%) for III and one (2.5%) for IV. Patients with p53-overexpressed tumours had a significantly higher number of tumour-bearing lymph nodes. There was no correlation of p53 expression with histological tumour types, second tumour recurrence and survival. Expression of p53 appears to be uncommon in patients with recurrent nodal metastases in NPC. It did not have prognostic value in this particular series of patients. PMID- 9393570 TI - The use of biliary CEA measurements in the diagnosis of recurrent colorectal cancer. AB - To assess the usefulness of biliary CEA determinations in the diagnosis of recurrent tumour, gallbladder bile was sampled in patients who underwent laparotomy for proven or suspected recurrent colorectal cancer and in control patients. Biliary CEA concentrations in controls were < 5 ng/ml, whereas significantly elevated CEA concentrations were found in the bile of all patients with tumour recurrence. Serum concentrations in these patients were elevated in 77% only. In a series of 12 patients with (a) suspicious lesion(s) on liver imaging but normal serum CEA concentration during follow-up, biliary CEA determination differentiated clearly between metastases and benign lesions. Biliary CEA determination seems to aid detection of tumour recurrence at an early stage and may preclude unnecessary surgery in patients with undefined liver lesions. PMID- 9393571 TI - Resection of pulmonary metastases from colorectal carcinoma. AB - A retrospective study was made on 22 patients who underwent surgery (28 operations) for lung metastases of colorectal origin from 1986 to 1995 at the Department of Surgery II, Padova University. The overall 5-year survival (OS) following pulmonary resection was 62% and the 5-year disease-free interval after metastasectomy (DFIM) 45%. The median survival was 23.6 months and the median DFIM 15.3 months. Univariate (Mantel Cox) and multivariate (Cox's model) analyses were used to identify any prognostic factors significant for OS and DFIM. Site and stage of primary colorectal carcinoma, number of pulmonary metastases at presentation, disease-free intervals between treatment of primary tumour and diagnosis of lung metastases (DFIP) appeared to have no influence on OS and DFIM. However, patients who underwent radical resection for metastases had a significantly longer DFIM than those who underwent 'non-radical' resections (P = 0.02), but radical resection had no significant positive effect on OS. A short DFIP, multiple and/or bilateral lesions, lung metastases occurring after liver resection with a curative aim are not contraindications to surgery in patients with pulmonary metastases from colorectal cancer, the main criterion for selection of patients being the possibility of performing 'radical' resection. PMID- 9393572 TI - Endocavitary Ir-192 radiation and laser treatment for palliation of obstructive rectal cancer. AB - Endoscopic laser therapy (ELT) either alone or combined with endocavitary Ir-192 radiation is performed for advanced, inoperable rectal cancer and when patients are ineligible for surgery due to severe concomitant medical illness. During the period from January 1984 to January 1997 we treated 81 patients (51 males, 30 females). Sixty-seven patients had ELT only using a ND-Yag Laser system. Twenty five patients (average age: 80.5 years) were ineligible for surgery (Group I). Forty-two patients (74.1 years) had an advanced locally inoperable tumour (Group II). Fourteen patients (76.5 years) underwent a combined therapeutic regime with endocavitary Ir-192 afterloading following ELT (Group III). Adequate desobliteration was achieved in 100% (groups I and III) and 97% (group II) of the patients. The average interval to aftertreatment was 8.4 weeks in group I and 9.4 weeks in group II, compared to 11.5 weeks in group III. Serious complications (perianal abscess, rectovaginal fistula) occurred in 3.7%, minor complications (laser-induced bleedings, unclear fever) in 12.3%. All laser-induced bleedings could be dealt with using laser therapy. The frequency of treatment was governed by tumour mass and the patient's survival. The results suggest that additional endocavitary radiation significantly prolongs the maintenance of normal bowel function compared with laser therapy alone. PMID- 9393573 TI - Lung carcinomas composed of rhabdoid cells. AB - Rhabdoid tumours form a distinctive morphological entity that is associated with aggressive biological behaviour. They have been described in several sites and tumour types. This paper presents three new cases of rhabdoid lung cancers. Lung cancers were analysed for the presence of cells with the rhabdoid phenotype: eccentric vesicular nuclei and abundant eosinophilic cytoplasm. Cells displaying this morphology were then subjected to immunohistochemistry and electron microscopy. The relevant clinical data on these cases were then accessed. Three cases conforming to the morphological, immunophenotypic and ultrastructural characteristics of rhabdoid cells were identified. Two of the cases were associated with foci of adenocarcinoma and the remaining case was a large cell neuroendocrine carcinoma. Two of the cases showed rapid clinical courses with the patients dying of disease within 6 months. Lung tumours with a rhabdoid phenotype are uncommon but are noteworthy because of their aggressive behaviour and, hence, poor prognosis. PMID- 9393574 TI - Carbon dioxide laser for cutaneous melanoma metastases: indications and limitations. AB - A total of 469 in-transit or satellite lesions were treated by carbon dioxide (CO2) laser vaporization in 15 patients. The treatment was performed mostly on an outpatient basis, under local anaesthesia. The technique was easily mastered and quickly performed. Wound healing and patient acceptance were good. A major drawback, however, proved to be the unexpected high incidence of recurrences at the lasered sites. In our opinion CO2 laser treatment may be considered as a palliative option in patients with a moderate to extensive amount of cutaneous metastases, whose lesions preferably are < 10 mm and in whom local excision is not feasible anymore. For extremity lesions this treatment may have a place after failure of isolated limb perfusion. CO2 laser treatment cannot be considered a first-line option unless the issue of local recurrences is solved. PMID- 9393575 TI - Tissue toxicity of doxorubicin in first and second hyperthermic isolated limb perfusion--an experimental study in dogs. AB - The aim of this experimental study in dogs was to assess the tissue toxicity of doxorubicin (DOX) and the impact of dose and pharmacokinetics after double isolated limb perfusion (ILP). Fifteen beagle dogs were assigned to three groups of five animals each. In the first ILP 0.75 mg/kg bodyweight (bw) DOX was given to all animals. In the second perfusion after an interval of 6 to 8 weeks the dosage was 0.5 mg/kg bw in group I, 0.75 mg/kg bw in group II, and 1.0 mg/kg bw in group III. At the same dosage tissue toxicity increased in comparison to the first ILP. At the second ILP there was a dose-toxicity relationship. At a dose of 0.75 mg/kg bw pharmacokinetics of DOX in the perfusate showed no significant differences between first and second perfusion. The mean muscle tissue levels during the second ILP were lower than during the first perfusion. However, in contrast to the first perfusion, they showed a further increase after perfusate eluation. A disturbed microcirculation caused by intima proliferations in arteries and arterioles fter the first ILP may impair the removal of DOX from the intravasal and interstitial compartment and can be assumed as a reason for increased tissue toxicity. Therefore, we recommend a reduction of DOX dose in the second ILP for clinical use. PMID- 9393576 TI - New technology in the analytical cell sciences: the laser scanning cytometer. AB - New technologies are making a major contribution to progress in applied clinical research in surgical oncology. The laser scanning cytometer is a new machine which combines the analytical capabilities of flow cytometry with the ability to inspect and visualize labelled cells and particles. This substantially reduces the uncertainty associated with assays in a wide range of surgical oncology research applications. This article introduces this new technology. PMID- 9393577 TI - Sister Joseph's nodule: a case report and review. AB - The history of Sister Joseph and a pathological review of the nodule named after her. PMID- 9393578 TI - Leiomyosarcoma of the renal vein. PMID- 9393579 TI - Biphasic sarcomatoid carcinoma of the lung: report of 5 cases and review of the literature. PMID- 9393580 TI - Lymph node recurrence of gallbladder carcinoma successfully managed by systemic chemotherapy with 5-fluorouracil and mitomycin C: report of a 5-year survivor. AB - Although gallbladder cancer (GBC) is believed to be chemoresistant, the effectiveness of chemotherapy against lymph node metastasis has been reported. We report a 70-year-old woman with advanced GBC in whom isolated, widespread lymph nodal recurrence after a radical resection responded completely to systemic chemotherapy with mitomycin C and 5-fluorouracil. This patient remains symptom free with no evidence of disease at 6 years after surgery (5 years after the initiation of chemotherapy). Both our case and a literature review suggest that nodal disease appears more chemosensitive than the primary lesion in GBC. Chemotherapy may provide long-term palliation for selected patients with isolated nodal recurrence. PMID- 9393581 TI - Haemangiopericytoma of the heart: report of a case with combined modality treatment. PMID- 9393582 TI - Cystic mesothelioma of the peritoneum. AB - A 48-year-old man presented with a 3-month history of weight loss and progressive right lower quadrant abdominal pain. His medical history was notable for appendectomy at age 17. Ultrasonography and computed tomography of the abdomen revealed a 12 cm multicystic mass in the right paracolic space. At laparotomy a large serous cyst was found arising from the lateral wall of the cecum, and four additional small cysts were found on the small bowel mesentery, greater omentum, liver capsule, and right hemi-diaphragm. Complete removal of the tumor was accomplished by right colectomy with extraperitoneal dissection of the large cyst and simple excision of the four smaller cysts. Final pathology with immunohistochemical staining confirmed cystic mesothelioma of the peritoneum. In this report we discuss the diagnostic workup and treatment of this rare disease. PMID- 9393583 TI - Plexosarcoma of the bladder. AB - Plexosarcomas are rare soft tissue sarcomas, previously reported in association with gastrointestinal autonomic nerve (GAN) plexi. We report the first case arising from the autonomic nerve plexus of the bladder. PMID- 9393584 TI - Is small tumour size alone enough for omitting axillary clearance? PMID- 9393585 TI - The administration of paclitaxel without prophylaxis for the prevention of hypersensitivity reactions: is this a rationale and safe therapeutic strategy? PMID- 9393586 TI - Differential expressions of cyclin A and the retinoblastoma gene product in histological subtypes of lung cancer cell lines. AB - Cell-cycle-dependent phosphorylation of the tumor-suppressor protein product of the retinoblastoma gene (RB) is mediated by a family of cyclin-dependent kinases and cyclins. We examined the expressions of RB protein and cyclin A protein in 13 small-cell lung cancer (SCLC) lines and 14 non-small-cell lung cancer (NSCLC) lines by immunoblotting. RB protein was not present or was of a mutant type in 77% of the SCLC lines (10/13) but was present in all the NSCLC lines. Cyclin A was expressed in 38% of the SCLC lines (5/13) and in 86% of the NSCLC lines (12/14). A positive correlation (P = 0.0034) between expression of cyclin A and wild-type RB protein was found by Fisher's exact probability test. Densitometric analysis of the expression of RB protein in RB(+) lung cancer lines showed that the phosphorylated form was predominant in 2/3 of the SCLC and 8/14 of the NSCLC lines. The positive correlation between the expressions of RB protein and cyclin A suggests that RB protein in most RB(+) lung cancer cell lines is a target of cyclin-A-dependent kinase and that the tumor-suppressor function may be inactivated by phosphorylation. PMID- 9393587 TI - Diagnostic and prognostic usefulness of N1,N8-diacetylspermidine and N1,N12 diacetylspermine in urine as novel markers of malignancy. AB - Recently, we found N1,N8-diacetylspermidine (Ac2Spd) and N1,N12-diacetylspermine (Ac2Spm) in human urine, and noted that their amount increased significantly in patients with urogenital malignancies. Previous findings that simultaneous reference to these diacetylpolyamines is useful in distinguishing cancer patients from healthy persons were confirmed by more recent analytical data on urine samples from several cancer patients. Further examination revealed that urinary Ac2Spm and Ac2Spd tended to decrease when cancer patients were treated and entered partial remission. In cases where the Ac2Spm and Ac2Spd levels were normal or near-normal after treatments, the prognosis of the patients was generally good. In contrast, when their level remained far above the normal limits after apparently effective treatment, the prognosis of the patients was poor. When a patient is in remission for more than 3 years, urinary levels of both Ac2Spm and Ac2Spd are stabilized and stay below the normal limits, with rare exceptions. The recurrence of a cancer as well as the complication of a second one during the period of follow-up examination was accompanied by elevation of urinary diacetylpolyamines. These observations indicate that urinary Ac2Spm and Ac2Spd are useful as prognostic indicators after treatment and during follow-up examination of cancer patients. PMID- 9393588 TI - Decreased levels of topoisomerase II alpha in human renal cell carcinoma lines resistant to etoposide. AB - Renal cell carcinoma (RCC) displays strong resistance against many chemotherapeutic drugs. Overexpression of P-glycoprotein (Pgp) appears to be part of this resistance. The involvement of another resistance mechanism, involving the decreased activity of DNA topoisomerase II (topoII), remains uncertain. By culturing the human RCC lines RC2 and RC21 in the presence of increasing concentrations of etoposide, we derived the variant sublines RC2E, RC21A and RC21E, that had acquired approximately 30-, 60- and 90-fold resistance to this drug respectively. RC2E, RC21A and RC21E were approximately 50-, 5- and 400-fold cross-resistant to doxorubicin respectively. RC2E and RC21E also showed cross resistance (approximately 200- and 3500-fold respectively) to vinblastine. Quantitative differences in MDR1 and Pgp expression (elevated in RC2E and RC21E) and topoII alpha (reduced in RC21E and RC21A) were demonstrated using Western blotting and the reverse transcriptase/polymerase chain reaction. Decreased amounts of topoII alpha were reflected in a reduced activity of RC21A and RC21E as measured by unknotting phage P4 DNA. Qualitative changes of the topoII alpha gene, such as point mutations in the motif B/DNBS and DNA-binding regions, or differences in methylation status of the promoter gene of RC21E, were not found. These cell lines represent a model of a solid tumor in which overexpression of Pgp, a combination of increased Pgp and decreased topoII alpha, and a decrease of topoII alpha are represented. PMID- 9393589 TI - A prospective study on the prognostic significance of urokinase-type plasminogen activator levels in breast cancer tissue. AB - Urokinase-type plasminogen activator (u-PA), which cleaves plasminogen to yield plasmin, is a serine protease of fibrinolysis and is presumed to play a key role in extracellular proteolysis and facilitate the migration of cancer cells. This study was conducted prospectively to evaluate the prognostic significance of u-PA antigen level in breast cancer tissues. u-PA concentrations in the cytosol of 226 breast cancer tissues were determined prospectively by enzyme-linked immunosorbent assay using cytosol fractions prepared for steroid hormone assay. The median follow-up period of the patients was 60 months. Various prognostic factors were evaluated by univariate analysis or multivariate analysis using the Cox proportional-hazards method. Patients with primary breast cancer containing high levels of u-PA had a significantly shorter disease-free survival than patients with low levels of u-PA antigens. In multivariate analysis, a high level of u-PA was an independent risk factor for disease-free survival, being independent of age, axillary node status, and estrogen receptor status. Among the major prognostic factors, a high u-PA antigen level, lymph node involvement, and a positive estrogen receptor status were the most important for predicting relapse-free survival (P = 0.044, P < 0.0001, P = 0.0039). This first prospective study confirmed the prognostic significance of the u-PA antigen level in association with other major prognostic factors. The results of our present study suggest that u-PA in breast cancer tissue might be involved in breast cancer invasion and metastasis. PMID- 9393590 TI - Leukaemia and lymphoma of the appendix presenting as acute appendicitis or acute abdomen. Four case reports with a review of the literature. AB - Leukaemic and lymphomatous infiltration of the appendix is rare and even rarer is acute appendicitis as the initial manifestation. From our routine biopsy material we collected four cases of haematological malignancies presenting as acute appendicitis or acute abdomen, caused or accompanied by tumoral infiltration of the appendix. Appendicitis was the initial manifestation that allowed diagnosis of the underlying disease. The clinical histories and histological examinations of the appendices and of one autopsy are described. We report the first detailed description of acute myeloid leukaemia involving the appendix, and three cases of lymphomatous infiltration of the appendix presenting with appendicitis, and give an overview of the literature. In these days of budgetary cuts in national health services, where one may be tempted not to have seemingly commonplace cases of appendicitis histologically verified, our cases emphasize that careful histopathological examination of all appendectomy specimens should be mandatory. Despite the fact that leukaemia and lymphoma of the appendix are rare, our cases illustrate that these must be included in the differential diagnosis of acute appendicitis and that physicians and surgeons have to be aware of these conditions. PMID- 9393591 TI - p53 nuclear immunoreactivity as a predictor of response and outcome following chemotherapy for metastatic bladder cancer. AB - p53 nuclear immunoreactivity was determined in primary bladder tumours from 50 patients who developed metastatic bladder cancer. We investigated the relationship between p53 nuclear immunoreactivity and the response and outcome following chemotherapy. p53 nuclear accumulation was detected in 48% of the primary tumours using the PAb1801 antibody in archival paraffin-embedded tissue sections. All patients received platinum-based combination chemotherapy including methotrexate for metastatic disease. The response to chemotherapy did not relate to the prevalence of p53 nuclear overexpression: 50% of the patients expressing p53 nuclear reactivity achieved a response compared to 27% of those without p53 expression (P = 0.14); overall, 38% of the patients responded. The median survival after chemotherapy was 5.9 months; 8.4 months for patients with p53 nuclear reactivity compared to 5.2 months for those without (P = 0.38). Multivariate analysis showed that performance status but not p53 nuclear status was an independent prognostic factor for survival following chemotherapy. The results indicate that patients with p53 nuclear immunoreactivity in the primary bladder tumour do not have a lower response rate or poorer outcome following chemotherapy for metastatic disease than do patients without p53 nuclear reactivity. PMID- 9393592 TI - Increased number of cancer cells in bronchial washing fluid detected by combining conventional cytology and high-resolution flow cytometry. AB - The present study was performed to improve early lung cancer diagnosis in bronchial washing fluid, thereby increasing the diagnostic sensitivity of bronchoscopy by means of high-resolution flow cytometry (FC). We combined dual parameter DNA/protein FC and conventional cytology in bronchial washing fluid samples from 112 patients with neoplastic and non-neoplastic lung diseases and found 43% of histologically confirmed tumor cases to be cytologically positive; 63% of the tumor samples were aneuploid, 52% of the aneuploid cases were cytologically positive and 48% were negative. In the negative cases, FC was an independent diagnostic factor. In 32% of the cases, FC also failed to detect abnormalities. However, the combination of both techniques increased the sensitivity in detecting neoplastic cells to 73%. Furthermore, simultaneous DNA/protein analysis allowed the recognition of aneuploid cell lines not detectable by single DNA measurement. Identification of aneuploid subpopulations by dual-parameter analysis in cytologically negative one-parameter FC "diploid" samples assumes an important diagnostic value. Dual-parameter DNA/protein FC is a valuable technique that increases the diagnostic yield of bronchoscopy with no risk for the patient and a low additional cost. PMID- 9393593 TI - Deficiency of neutrophilic granule membrane glycoproteins in the myelodysplastic syndromes: a common deficiency in 216 patients studied by the Cancer and Leukemia Group B. AB - Previous studies on neutrophils in patients with the myelodysplastic syndromes (MDS) have indicated deficiencies in the contents of primary and secondary granules. However, the granule membrane remains virtually unstudied despite its essential role in the dynamic function of the cytoplasmic granules. In this study, we examined the membrane glycoproteins of primary and secondary granules of peripheral blood and/or bone marrow neutrophils using the monoclonal antibody H36/71 to CD15 glycoproteins. In addition, myeloperoxidase activity and antigen, elastase and lactoferrin were also studied using cytochemical and immunocytochemical stains. A total of 216 patients were included. Deficiencies of granule membrane glycoproteins were the most common, detected in 49%, followed by myeloperoxidase activity (17%), elastase (16%), myeloperoxidase antigen (9%), and lactoferrin (8%). Multiple deficiencies always included granule membrane deficiency. We conclude that granule membrane defects are common in MDS, may provide a common mechanism for multiple granule deficiencies, and may prove to be an additional abnormality associated with granulocyte dysfunction. PMID- 9393594 TI - Synthesis of quantitation standards for nested competitive PCR for the determination of minimal residual disease in B-lineage acute lymphoblastic leukaemia. AB - Quantitative PCR-based assays of leukaemic cells in remission marrow hold promise for therapeutic guidance, but are not yet sufficiently reliable for clinical application. For B-lineage ALL, these assays usually involve PCR of clonal somatic gene rearrangements of the immunoglobulin heavy chain gene. The most accurate quantification can be achieved by competitive PCR. Here we present a novel approach for the production of reference standards for use in nested competitive PCR of these gene rearrangements, which might enable more reliable assessment of MRD for prognosis and selection of patients for individualised therapy. PMID- 9393596 TI - Allelic loss in childhood acute lymphoblastic leukemia. AB - Acute lymphoblastic leukemia (ALL) is the most frequent cancer encountered in children. Little is known about the molecular basis of childhood ALL, although the clinical, pathological, and immunophenotypic features have been well documented. To understand the role of tumor suppressor genes (TSGs) in the development of this disease, we performed a detailed allelotype analysis. Twenty nine patients (24 pre-B and 5 of T-cell lineage) were investigated for loss of heterozygosity (LOH), using 49 highly polymorphic markers distributed over 13 chromosomal arms which are known or postulated to contain TSGs. The highest rates of allelic losses were observed in chromosomes 9p and 12p which were deleted in 29 and 32% of the informative patients, respectively. These are among the most frequent alterations found in childhood ALL. Other losses were found at a lower frequency in chromosomes 6p, 6q, 9q, 17p, and 17q. No LOH was found at chromosomes 3p, 5q, 11p, 11q, 13q and 18q in any patient. These results suggest that many TSGs may be involved in the development of childhood ALL. PMID- 9393595 TI - Salvage therapy for relapsed or refractory childhood acute lymphocytic leukemia by alternative administration a lymphoid- and myeloid-directed chemotherapeutic regimen consisting of dual modulation of ara-C, hydroxyurea, and etoposide. AB - Risk-directed chemotherapeutic regimens in recent use have improved the prognosis of children with acute lymphocytic leukemia (ALL). However, many patients relapse during or shortly after cessation of the initial continuation chemotherapy. Since achievement of a second complete remission (CR) is the initial step in successful retreatment effort, it is important to develop salvage protocols for children with relapsed or refractory ALL. In the present study, we developed a new salvage protocol (MLL-93) and applied the concept of dual chemical modulation of cytarabine, hydroxyurea, and etoposide with the alternative administration of high doses of myeloid- and lymphoid-directed agents. We also planned to perform allogeneic bone marrow transplantation (BMT) following a CR if patients had HLA identical donor(s). The six patients treated with the MLL-93 protocol achieved a second CR. One patients in CR died of interstitial pneumonia after an unrelated allogeneic BMT. The other five patients have been in CR for 12-41 months. We suggest that the concepts of alternative administration of lymphoid- and myeloid directed drugs and biochemical modulation are useful in the treatment of children with relapsed or refractory ALL. PMID- 9393597 TI - Quantification of P-glycoprotein in chronic lymphocytic leukemia by flow cytometry. AB - The P-glycoprotein (P-gp) was investigated in 40 patients with chronic lymphocytic leukemia by immunological, functional and quantitative assays. The proportion of positive cases with the anti-Pgp McAb UIC2 was 30% and increased to 64% after neuraminidase treatment (p = 0.002). Fifty-six per cent of cases were positive with the functional test with rhodamine 123 and verapamil. A negative correlation was found between the number of cells stained with the McAb and the functional test (p = 0.006). The mean of P-gp molecules was 2509 +/- 2805 molecules per cell; these values were higher than in the control K562 cell line in the majority of cases. The number of positive cases and P-gp molecules were higher in treated than in untreated patients (p = 0.01 and 0.07). There were no significant differences with respect to response to treatment, but a higher number of P-gp molecules was found in non-responders. When the results of the functional test were put together with the quantification assay this allowed the detection of 71% non-responders. Our findings suggest that quantification of the P-gp could be of value in the assessment of possible drug resistance in CLL. PMID- 9393598 TI - C/EBP-epsilon: chromosomal mapping and mutational analysis of the gene in leukemia and preleukemia. AB - We and others have cloned a novel human gene CCAAT/enhancer-binding protein epsilon (C/EBP-epsilon) encoding a member of the C/EBP gene family. It is exclusively expressed in myeloid and T-lymphoid cells and appears to have an important role in inducing expression of several myeloid-specific genes. We used a polymerase chain reaction (PCR)-based technique to examine DNA from 93 hamster/human radiation hybrid clones in order chromosomally to map C/EBP-epsilon to 14q11.2 (between D14S264 and D14S275) which is telomeric to the T-cell receptor alpha and delta genes and centromeric to several other myeloid gene products including Cathepsin G (CTSG) and Chymase-1 (CMA1). To determine whether C/EBP-epsilon behaves as an altered tumor-suppressor gene, samples from patients with acute myelogenous leukemia (AML) and myelodysplastic syndrome (MDS) evolving to AML were studied for loss of heterozygosity (LOH) using microsatellite sequences that we identified within 0.2 kb of the amino-terminus of the human C/EBP-epsilon gene. Allelic loss of the C/EBP-epsilon gene was detected in four out of 20 (20%) evolving MDS cases and in none of the 17 AML and 17 T-cell leukemia cases. Mutational analysis of the gene was performed using PCR-SSCP on 37 AML and 40 MDS cases including those with LOH at the gene. No abnormalities were found suggesting that the altered gene in this region is not C/EBP-epsilon. Also, C/EBP-epsilon was examined by Southern blot analysis on DNA samples from 20 AML patients and 10 AML cell lines. No rearrangements or amplifications of the gene were detected. Taken together, we have mapped C/EBP-epsilon to 14q11.2, a region containing other myeloid and T-lymphoid specific genes. Furthermore, no structural alterations were detected in the C/EBP-epsilon gene. PMID- 9393599 TI - Differential phosphorylation of lamin B2 in normal and leukemic cells. AB - Lamins constitute the nuclear lamina, which underlie the inner membrane of the cell nucleus. Phosphorylation of lamins is a key factor in the regulation of nuclear structure during the cell cycle and of gene transcription. Since an uncontrolled cell cycle and altered gene transcription are major characteristics of neoplasms, we looked for differences in lamin B2 phosphorylation between PBMC, ALL and AML cells. Using different lamin B2-specific antibodies, we detected two different lamin B2 species termed lamin B2 and B2A. Although phosphorylation of lamin B2 in leukemic cells was reminiscent of resting cells, the majority of ALL and AML samples showed significantly higher and more altered lamin B2A phosphorylation compared to PBMC. It remains to be elucidated which mechanism leads to these alterations and whether it could explain the extended G1-phase frequently observed in ALL cells. PMID- 9393600 TI - Growth factor stimulation of hematopoietic cells leads to membrane translocation of AKT1 protein kinase. AB - AKT1 is the human homolog of the v-akt oncogene. AKT1 has two distinct protein domains, one serine/threonine kinase domain and one pleckstrin homology (PH) domain. We studied the expression and activity of AKT1 in hematopoietic cell lines. The expression of AKT1 was constitutive in hematopoietic cells of various stages of development. In the growth factor dependent MO7e cells, serum and growth factor starvation resulted in an early 50% fall in activity which was maintained over 24 h. Treatment of cells which growth factors or agents which induce differentiation activated AKT1. The subcellular localization of AKT1 in MO7e cells was altered as it was activated. High AKT1 kinase activity was associated with membrane fractions in stimulated cells, in contrast to the much lower AKT1 activity in membranes of cells starved of serum and growth factor for 1 h. These results demonstrate AKT1 kinase activity and its regulation by extracellular signaling factors in vivo in hematopoietic cells, and suggest that the activation of AKT1 involves intracellular translocation of the kinase from cytosol to membrane. PMID- 9393601 TI - The influence of recombinant human tumor necrosis factor alpha and its muteins used alone or in combination with 2-chlorodeoxyadenosine on normal and leukemic hematopoiesis in vitro. AB - We investigated the influence of TNF alpha and its muteins III, V and VI on the colony growth of normal and CML CFU-GM cells as well as on CFU-L clonogenic blasts from AML patients in cultures in vitro. The muteins were constructed using a synthetic cDNA fragment substituting nucleotides coding for the N-terminal amino acids in the chain of native TNF alpha. We observed that TNF alpha and mutein VI inhibited the growth of colonies formed by both types of CFU-GM and CFU L cells in a greater degree than muteins III and V. In addition, the percentage of colony growth inhibition after the use of mutein VI was the greatest. These observations were the basis for the evaluation of interaction between 2-CdA and TNF alpha and between 2-CdA and mutein VI. We have confirmed that 2-CdA used together with mutein VI acts synergistically on CFU-GM and CFU-L cells, whereas 2 CdA and TNF alpha show the additive effect. PMID- 9393602 TI - Effect on cell kill of addition of multidrug resistance modifiers cyclosporin A and PSC 833 to cytotoxic agents in acute myeloid leukaemia. AB - Multidrug resistance (MDR) mediated by the drug efflux pump P-glycoprotein (Pgp), may cause remission failure and relapse in patients with acute myeloid leukaemia (AML) by extruding cytotoxic agents such as anthracyclines from leukaemic cells thus allowing them to survive. Cell line data suggest that reversal of MDR is possible using modifying drugs such as cyclosporin A (CSA) and its analogue PSC 833. We have investigated the effects on cell kill of the addition of CSA and PSC 833 to daunorubicin, idarubicin, mitozantrone, etoposide and cytarabine in 52 fresh cell samples from AML patients using an MTT assay. Pgp status was determined by using monoclonal antibodies JSB-1 and MRK-16 and by assessment of rhodamine efflux. Although overall each cytotoxic-modifier combination produced significant improvements in cell kill compared to cytotoxic alone (P values ranged from P < 0.001 to P = 0.017), modifiers also produced significant cytotoxicity in their own right, and no consistent difference was seen between responses in Pgp-positive and negative groups. Up to one in three Pgp-positive samples failed to show any improvement in cell kill with the addition of CSA or PSC 833, possibly owing to co-expression of alternative resistance mechanisms not affected by the MDR modifiers. The best responses were seen when PSC 833 was added to idarubicin, with 7 out of 22 Pgp-positive cases (32%) showing five-fold improvements in cell kill or better compared to idarubicin alone. Comparison of equimolar concentrations of the two modifiers in the Pgp positive group failed to show a significant difference in cell kill, though PSC 833 was markedly superior to CSA in a minority of highly responsive samples which demonstrated clear evidence of MDR reversal. Our in vitro data suggest that MDR modifiers such as CSA and PSC 833 could play an important role in the therapy of AML and indicate the need for prospective randomised trials to assess their clinical efficacy. PMID- 9393603 TI - Bufalin reduces the level of topoisomerase II in human leukemia cells and affects the cytotoxicity of anticancer drugs. AB - When human leukemia HL-60 cells were treated with 10(-7) M bufalin, the amounts of both topoisomerase (topo) II alpha and II beta and the activity of topo II decreased markedly and were almost undetectable 18 h after the start of treatment. The level of topo II mRNA started to decrease immediately after the start of treatment with bufalin, with a subsequent decrease in the amount of topo II alpha protein. These changes preceded the fragmentation of DNA, a typical feature of apoptosis. The results suggest that bufalin caused a marked decrease in the steady-state level of topo II alpha mRNA, which led to a decrease in the amount and activity of the enzyme and to the induction of apoptosis. A reduction in the level of topo II alpha by bufalin was also observed in other lines of human leukemia cells such as ML1 and U937. The results were exploited to potentiate the effects of cisplatin and retinoic acid (RA) on HL-60 cells: pretreatment of HL-60 cells with 10(-7) M bufalin for 6 h increased the inhibitory effects of cisplatin and RA on cell growth and enhanced the induction of cell death. PMID- 9393604 TI - A case-control study of reproductive factors and risk of lymphomas and myelomas. AB - The relationship between reproductive factors and risk of lymphoid neoplasms was investigated in a hospital-based case-control study conducted in northern Italy on women with histologically confirmed incident Hodgkin's disease (HD) (n = 68), non-Hodgkin's lymphomas (NHL) (n = 180) and multiple myelomas (MM) (n = 71), and 448 controls admitted to hospitals, for acute, non-neoplastic, non-immunological and non-gynecological conditions. The odds ratios (OR) of HD were 0.6 for > or = 3 pregnancies compared to nulligravidae, and 0.5 for > or = 1 total (spontaneous and induced) abortions compared to women reporting no abortions. Compared to nulliparae, the OR of HD was 0.9 in parae and 0.3 in those with first birth when aged < 20 years. The OR of NHL and MM in relation to number of pregnancies, abortions and births, age at first birth and time since last birth were close to unity. Results were similar for the relation between reproductive factors and HD in women younger than 50 years. The OR of NHL was above unity (OR 2.2, 95% CI 1.0 to 4.9) for women aged < 50 years reporting one or more pregnancies as compared to nulliparae, and for women reporting the last birth since less than 10 years (OR 2.9, 95% CI 1.1 to 7.4). Early events in pregnancy, including changes in immunological status, rather than exposure to female sex hormones are likely mechanisms for the protection of pregnancies and abortions on the risk of HD. PMID- 9393605 TI - Chronic myelomonocytic leukemia in a patient with antiphospholipid syndrome: first case report. PMID- 9393606 TI - Continuous hemofiltration in the management of 'retinoic acid syndrome'. PMID- 9393607 TI - Lymphocytes with cerebriform nuclei in chronic B-cell lymphoproliferative diseases. AB - Two patients with chronic lymphoproliferative diseases, a splenic marginal zone lymphoma and hairy cell leukaemia variant, were reported. In both diseases a B immunophenotype was demonstrated but a variable percentage of lymphoid cells (30 and 52%) showed a highly irregular nuclear outline, sometimes mimicking the nuclei of Sezary cells. PMID- 9393608 TI - Secondary amyloidosis in the course of idiopathic myelofibrosis. AB - Secondary amyloidosis without a known cause, diagnosed antemortem in a patient with idiopathic myelofibrosis, is reported here. This is the first such case, to our knowledge. Amyloid deposits were seen in the bone marrow, renal glomeruli and jejunum. Reasons for investigating for amyloidosis were hypogammaglobulinemia, proteinuria and recurrent diarrhea. PMID- 9393609 TI - Indo-French symposium on apoptosis and multidrug resistance. PMID- 9393610 TI - Detection of mineral-dust-induced DNA damage in two mammalian cell lines using the alkaline single cell gel/comet assay. AB - It has been estimated that over three million workers in the USA are potentially exposed to silica or other mineral dusts. Results of epidemiological studies evaluating whether silica or glass fibers increase lung cancer risk to the exposed workers are inconclusive. Detection of DNA damage in cells exposed to genotoxic agents is being used to assess the carcinogenic potential of environmental agents. The alkaline (pH > 13) single cell gel/comet (SCG) assay was used to determine and compare DNA damage in cultured Chinese hamster lung fibroblasts (V79 cells) and human embryonic lung fibroblasts (Hel 299 cells) exposed to crystalline silica (Min-U-Sil 5), amorphous silica (Spherisorb), carbon black, and glass fibers (AAA-10). V79 or Hel 299 cells were exposed to these mineral dusts for 3 h at various concentrations. Min-U-Sil 5 and AAA-10, at almost all concentrations tested, caused a significant increase in DNA migration measured as tail length in both V79 and Hel 299 exposed cells. However, the increase was much higher in V79 then in Hel 299 cells for Min-U-Sil 5. Tail length was also increased relative to controls after amorphous silica treatment, but not to the same extent as that induced by crystalline silica. Exposure to carbon black did not induce DNA migration at any of the concentrations tested. These results indicate that silica and glass fibers, but not carbon black, can induce DNA damage in mammalian cells, and that crystalline silica has a higher DNA-damaging activity than amorphous silica. For glass fibers, induction of DNA damage in both V79 and Hel 299 cells was observed even at a concentration 10 times lower than silica and the response was similar in both cell lines. These results suggest that the SCG/comet assay is useful for the detection of DNA damage caused by occupationally related dusts/particles. PMID- 9393611 TI - Genetic toxicity studies of 1,2,3,4-tetrahydro-9-acridinamine (tacrine). AB - The mutagenicity and clastogenicity of 1,2,3,4-tetrahydro-9-acridinamine (tacrine) were studied in vitro using the Salmonella mutagenicity test and the induction of chromosome aberrations in Chinese hamster ovary (CHO) cells, and in the mouse bone marrow micronucleus test in vivo. This chemical is currently being used to treat dementia arising from Alzheimer's Disease. Tacrine was mutagenic in Salmonella but did not produce chromosome damage in CHO cells or in mouse bone marrow cells. A clear mutagenic response was seen in strain TA97 with rat and hamster liver S9; inconsistent results were obtained without S9. No mutagenicity was seen in strains TA98 and TA100 without S9, and inconsistent results were seen with S9. There was no induction of chromosome aberrations in cultured CHO cells with or without S9. Oral administration to mice of tacrine daily for three days did not result in the induction of micronuclei in their bone marrow cells. The mutagenic response in Salmonella, and the structure of the molecule, suggests that tacrine may be carcinogenic when tested in rodents. This information must be considered when preparing benefit-risk determinations for medical uses of this substance. PMID- 9393612 TI - Benzidine-DNA adduct levels in human peripheral white blood cells significantly correlate with levels in exfoliated urothelial cells. AB - In a cross-sectional study of 33 workers exposed to benzidine and benzidine dyes and 15 non-exposed controls, we previously reported that exposure status and internal dose of benzidine metabolites were strongly correlated with the levels of specific benzidine-DNA adducts in exfoliated urothelial cells. We also evaluated DNA adduct levels in peripheral white blood cells (WBC) of a subset of 18 exposed workers and 7 controls selected to represent a wide range of adducts in exfoliated urothelial cells. Samples were coded and then DNA was analyzed using 32P-postlabeling, along with n-butanol extraction. One adduct, which co chromatographed with a synthetic N-(3'-phospho-deoxyguanosin-8-yl)-N' acetylbenzidine standard, predominated in those samples with adducts present. The median level (range) of this adduct in WBC DNA was 194.4 (3.2-975) RAL x 10(9) in exposed workers and 1.4 (0.1-6.4) in the control subjects (p = 0.0002, Wilcoxon Rank Sum Test). There was a striking correlation between WBC and exfoliated urothelial cell adduct levels (Pearson r = 0.84, p < 0.001) among exposed subjects. In addition, the sum of urinary benzidine, N-acetylbenzidine and N,N' diacetylbenzidine correlated with the levels of this adduct in both tissues. This is the first study in humans to show a relationship for a specific carcinogen adduct in a surrogate tissue and in urothelial cells, the target for urinary bladder cancer. PMID- 9393613 TI - Induction of micronuclei in human, goat, rabbit peripheral blood lymphocytes and mouse splenic lymphocytes irradiated in vitro with gamma radiation. AB - The frequencies of gamma-ray-induced micronuclei (MN) in cytokinesis-blocked (CB) lymphocytes at several doses were measured in three donors of four species (human, goat, rabbit, mouse). Measurements performed after irradiation showed a dose-related increases in MN frequency in each of the donors studied. The relative sensitivity of mouse in spleen lymphocytes (SLs), goat in peripheral blood lymphocytes (PBLs) and rabbit PBLs compared with human PBLs was estimated by best fitting linear-quadratic model based on the radiation-induced MN data over the range from 0 to 400 cGy. In the case of MN frequency with 0.2, the relative sensitivities of mouse SLs, goat PBLs and rabbit PBLs were 1.67, 0.98 and 0.39, respectively. These data indicate that the induction of MN in CB cells following irradiation is similar in human and goat PBLs, and PBLs from rabbit were much less sensitive to the MN induction effects of gamma-radiation than those from human. Compared with the radiation-induced MN formation in the PBLs of human, the SLs of mouse were more radiosensitive. PMID- 9393614 TI - Differential induction of adaptive responses by paraquat and hydrogen peroxide against the genotoxicity of methyl mercuric chloride, maleic hydrazide and ethyl methane sulfonate in plant cells in vivo. AB - Induction of adaptive response by conditioning doses of paraquat (PQ) and hydrogen peroxide (H2O2) in embryonic shoot cells of Hordeum vulgare and root meristem cells of Allium cepa was tested against the genotoxicity of challenge doses of methyl mercuric chloride (MMCl), maleic hydrazide (MH) or ethylmethane sulfonate (EMS). Plant tissue fixed at different recovery hours following the challenge treatments was analysed for cells with genotoxicity markers that include spindle or chromosome aberrations and micronuclei. The results provided clear-cut evidence that whereas H2O2 induced adaptive response for the chromosome damage caused by MMCl and MH, PQ induced the same for MMCl and EMS, but not for damage caused by MH. The findings pointed to the differences in the underlying mechanisms of oxidative responses induced by H2O2 and O2-. PMID- 9393615 TI - Quercetin and myricetin protect against hydrogen peroxide-induced DNA damage (strand breaks and oxidised pyrimidines) in human lymphocytes. AB - The effects of the flavonoids quercetin and myricetin, and the antihepatotoxic agent silymarin, on hydrogen peroxide-mediated DNA damage in human lymphocytes were determined using alkaline single-cell gel electrophoresis (the comet assay). Treatment with hydrogen peroxide increased the levels of DNA strand breaks and oxidised pyrimidine bases in these cells. Quercetin was protective at concentrations above 10 microM and myricetin decreased oxidant-induced DNA strand breakage at concentrations of 100 microM. Cellular metabolism may alter the antioxidant efficacy of the flavonoids. Silymarin had no protective effect at any of the concentrations tested. None of these flavonoids was itself genotoxic. Neither alpha-tocopherol nor beta-carotene decreased hydrogen peroxide-induced DNA breakage. The differences in effectiveness of these dietary compounds against oxidative DNA damage may be explained by differences in their chemical structure or location within the cell. PMID- 9393616 TI - Construction of mutants of Salmonella typhimurium deficient in 8-hydroxyguanine DNA glycosylase and their sensitivities to oxidative mutagens and nitro compounds. AB - 8-Hydroxyguanine (8-OH-G) DNA glycosylase is an enzyme involved in repair of oxidative DNA damage, e.g., 8-OH-G in DNA. In order to assess the roles of 8-OH-G in spontaneous and chemically-induced mutagenesis, the mutMST gene encoding 8-OH G DNA glycosylase of Salmonella typhimurium was disrupted in several Ames tester strains, i.e., S. typhimurium TA1535 (hisG46, uvrB-, rfa), TA1975 (hisG46, uvr+, rfa) and TA102 (hisG428, uvr+, rfa). The spontaneous mutation frequencies were increased 2.4 and 1.6 times, respectively, by the mutMST deletions in strains TA1535 and TA1975, which are spontaneously reverted to His+ by mutations mainly at G:C base pairs. The resulting strains YG3001 (TA1535 delta mutMST) and YG3002 (TA1975 delta mutMST) were 2 to 8 times more sensitive to the mutagenicities of methylene blue plus visible light, neutral red plus visible light and 2 nitrofluorene than the parent strains. The strain YG3002 but not YG3001 was about 30 times more sensitive to the mutagenicity of 4-nitroquinoline N-oxide than the parent strain TA1975. Neither hydrogen peroxide nor phenazine methosulfate was mutagenic in the mutMST-deletion strains as well as in the parent strains. In contrast, the mutMST deletion did not affect the spontaneous mutation frequency of strain TA102, which has an A:T base pair at the critical site for reversion. The sensitivities of strain TA102 to the chemicals were not enhanced by the mutMST deletion except for hydrogen peroxide. These results suggest that 8-OH-G in DNA plays important roles in spontaneous mutagenesis occurring at G:C base pairs in S. typhimurium, and some nitro aromatics such as 4-nitroquinoline N oxide or 2-nitrofluorene as well as the photosensitizers plus visible light can produce 8-OH-G in DNA, thereby inducing mutations. In the case of 4 nitroquinoline N-oxide, 8-OH-G rather than DNA adducts seems to play major roles in mutagenesis in uvr+ background. The new strains could be useful for the evaluation of the roles of 8-OH-G in mutagenesis in S. typhimurium and permit the efficient detection of some oxidative mutagens in the environment. PMID- 9393617 TI - Metabolism of galangin by rat cytochromes P450: relevance to the genotoxicity of galangin. AB - The mutagenicity of flavonols seems to depend on the number and position of hydroxyl groups in the B ring. Galangin is a flavonol that does not have any hydroxyl group in the B ring and has been suggested to be a substrate of cytochromes P450 which, through the hydroxylation of the B ring, could metabolise it to more genotoxic products. The present study was undertaken to test this hypothesis. Using high performance liquid chromatography we show that glangin is sequentially transformed to kaempferol and then to quercetin by a mechanism dependent on cytochrome P450 reactions. The metabolites of galangin are responsible for its mutagenicity in Salmonella typhimurium reversion assay and for the induction of chromosomal aberrations in V79 cells. PMID- 9393618 TI - Sister chromatid exchange analysis in patients exposed to low dose of iodine-131 for thyroid scintigraphy. AB - To determine the genotoxic risk associated with diagnostic exposure to low doses of iodine 131 (131I), sister chromatid exchange (SCE) analysis was performed in lymphocytes of 18 non-smoking women who received 370 kBq (10 microCi) intravenous 131I sodium iodide as an adjuvant for scintigraphy for diagnosing thyroid nodularity. SCE frequencies were measured before and after 131I administration. SCE results in the pre-treated phase were regarded as control. Although SCE values 24 h after 131I administration did not show a significant increment (p > 0.05), there was a significant increase 72 h after treatment (p < 0.05). These results indicate that genetic damage might be induced by low dose of 131I. PMID- 9393619 TI - Evaluation of the mutagenicity of acetochlor to male rat germ cells. AB - Male rat dominant lethal (DL) assays conducted on the herbicide acetochlor are described. Single dose studies conducted at the maximum tolerated dose (MTD, < or = 1000 mg/kg) produced no effects on any of the DL assay parameters at any of the ten weekly sampling periods. It is concluded that acetochlor is non-mutagenic to rat germ cells. Due to initial limited knowledge of the MTD of acetochlor it was also evaluated in the DL assay at a dose level of 2000 mg/kg. At this high dose level severe bodyweight loss and some deaths occurred among the treated animals. In addition, reduced implantations and reduced pregnancy rates were observed at the third sampling period (18-25 days post dosing) in the absence of an increase in early post-implantation deaths. These results indicated that the use of supra MTD doses of acetochlor had reduced the fertility of the treated males leading to the production of a pseudo-DL assay response, as alerted to and defined by Ehling. Although several such pseudo-DL assay responses have been described, none have been explained mechanistically. It was therefore decided to pursue the effects seen in the DL assay when using supra-MTD doses of acetochlor. Ova analysis of female rats mated with male rats exposed to 2000 mg/kg acetochlor revealed unfertilized ova at the critical third sampling time. Normal fertilization of ova was observed at the first and fifth sampling period and, for a dose of 200 mg/kg acetochlor, at the third sampling period. The magnitude and temporal nature of these effects confirmed the induction of a pseudo-DL assay response, and studies were then undertaken to probe its genesis. Rats treated with 2000 mg/kg acetochlor had normal testicular and epididymal pathology and normal sperm numbers and sperm motility at the critical third sampling period. Despite a small reduction in testicular and epididymal glutathione levels 12 h after exposure to 2000 mg/kg acetochlor, testicular LDH and LDH-X enzyme levels were unaffected. Further, no reduction in the level of free sulphydryl groups ( SH) were observed in epididymal caput sperm heads isolated 0.5, 7 or 14 days after treatment of male rats with 2000 mg/kg acetochlor. The only sperm parameter affected by treatment with 2000 mg/kg acetochlor was an increase in epididymal cauda sperm with head abnormalities. The non-specific nature of this effect was considered inadequate to explain fully the high dose fertility effects seen in the DL assays, which therefore remain unexplained. The present data establish that acetochlor is non-mutagenic to rat germ cells. They also confirm the importance of segregating mutagenic and fertility effects in the DL assay, and emphasize the need for appropriate dose-setting studies prior to the conduct of rodent genetic toxicity assays. PMID- 9393620 TI - Chromosomal aberrations and micronuclei in lymphocytes of workers at a phosphate fertilizer factory. AB - The frequencies of chromosomal aberrations (CA) and micronuclei (MN) in peripheral blood lymphocytes of 40 workers at a phosphate fertilizer factory in North China, were studied. HF and SiF4 are the main air pollutants and small amounts of dust containing fluoride, NH3 and SO2 were also present in the factory. It was shown that the chemicals caused an increase in both CA and MN. The mean frequencies per 100 metaphase of major CA type (chromosome rings, translocations, and dicentrics) of the workers and the non-exposed controls were 0.91 and 0.24 (p < 0.01), respectively. The average percentages of lymphocytes with MN of the workers and the controls were 1.55 +/- 0.71 and 0.62 +/- 0.54 (p < 0.01), respectively. Both CA frequency and MN frequency of the workers increased with length of the chemical exposure period up to 10 years. PMID- 9393621 TI - Lack of emodin genotoxicity in the mouse micronucleus assay. AB - The study was performed to investigate the potential of emodin (1,3,8-trihydroxy 6-methylanthraquinone) to induce micronuclei in polychromatic erythrocytes (PCEs). Mice of both genders received a single oral dose of 2000 mg emodin/kg and were killed 24 and 48 h later. Bone marrow cells were collected from 5 males and 5 females and 2000 PCEs per animal were scored for the presence of micronuclei. There was no enhancement in the frequency of micronuclei at both preparation intervals when compared to the negative controls. Blood level examinations confirmed the systemic availability of emodin. Plasma levels of up to 190 micrograms emodin/ml represented concentrations being in the concentration range that induced positive responses in several genotoxicity cell culture assays. PMID- 9393622 TI - Development of a novel CHL/IU cell line with an incorporated gpt shuttle vector for concurrent analysis of gene mutations and chromosome aberrations. AB - A cosmid shuttle vector containing the target gene of Escherichia coli gpt coding xanthine-guanine phosphoribosyl transferase was constructed. The shuttle vector was designed to be rescued into the gpt-deficient Escherichia coli from Chinese hamster CHL/IU cells through an in vitro packaging method. Mutations occurred at the target gene can be detected with a selective agent, 6-thioguanine (6-TG). The shuttle vector was stably transfected into CHL/IU cells to give several cell lines containing copies of the shuttle vector in the chromosomes. Each cell line exhibited a characteristic rescue efficiency (0 to 1.9 x 10(5) CFU/microgram of genomic DNA) of the shuttle vector and spontaneous mutation frequency (3.9 x 10( 5) to over 10(-2)) at the 6-TG selection. One transgenic cell line (KN63), which showed a higher rescue efficiency and a low spontaneous mutation frequency, was selected and tested for the ability to respond to a genotoxic agent, N-methyl-N' nitro-N-nitrosoguanidine (MNNG). MNNG increased both the mutation frequency at the target gene and the number of the cells with chromosome aberrations. DNA sequence analysis of 6-TG mutants showed that predominant mutations (10/14) were identified as G:C to A:T transitions in MNNG-induced mutants, whereas transversions were predominant (5/9) in spontaneous mutants. These results suggest that this transgenic CHL/IU cell line can be a useful tool for analyzing the relation between gene mutations and chromosome aberrations. PMID- 9393623 TI - Suppression of aflatoxin B1- or methyl methanesulfonate-induced chromosome aberrations in rat bone marrow cells after treatment with S-methyl methanethiosulfonate. AB - The suppressive effect of S-methyl methanethiosulfonate (MMTS) on aflatoxin B1 (AFB1)- or methyl methanesulfonate (MMS)-induced chromosome aberrations (CA) in rat bone marrow cells was studied. MMTS significantly suppressed CA induced by both AFB1 (an indirect-acting carcinogen) and MMS (a direct-acting carcinogen). Suppression was observed at all periods (6, 12, 18, 24 and 48 h) after AFB1 or MMS treatment and in all doses of AFB1 (5, 10 and 20 mg/kg) or MMS (50, 75 and 100 mg/kg) investigated. AFB1-induced CA was potently suppressed by MMTS given between 2 h before and 6 h after the AFB1 injection. The suppression of AFB1 induced CA by MMTS paralleled the dose of MMTS when MMTS was given in a dose range of 1-20 mg/kg body weight. MMS-induced CA was potently suppressed by MMTS given between 2 h before and 2 h after the MMS injection. The suppressive effect of MMTS on MMS-induced CA paralleled the dose of MMTS when MMTS was given in a dose range of 1-15 mg/kg body weight. Diphenyl disulfide, which modifies -SH groups in proteins like MMTS, also significantly suppressed both AFB1- and MMS induced CA. Although other mechanisms are not excluded, the suppression of carcinogen-induced CA by MMTS may result from the ability of MMTS to modify -SH groups in proteins. The juices of cabbage and onion, which contain considerable amounts of MMTS and S-methyl-L-cysteinesulfoxide (the precursor of MMTS), also significantly suppressed AFB1- or MMS-induced CA. These results suggest that MMTS is a possible chemopreventive agent against cancer. PMID- 9393624 TI - The Nobel Lectures in Immunology. The Nobel Prize for Physiology or Medicine, 1996. Cellular immune recognition and the biological role of major transplantation antigens. PMID- 9393625 TI - Tumour immunology: alternative perspectives. PMID- 9393626 TI - A novel mycobacterial antigen relevant to cellular immunity belongs to a family of secreted lipoproteins. AB - The gene sequence of a novel 24.1 kDa Mycobacterium tuberculosis protein was identified within the Sanger Centre (UK) M. tuberculosis genome database (cosmid MTCY24G1) by searching with a 126 bp DNA sequence isolated from a genomic M. leprae lambda gt11 library with M. leprae reactive human T cell clones as probes. The 24.1 kDa antigen is common to the vaccine strain Mycobacterium bovis BCG, as well as Mycobacterium leprae. The 699 bp open reading frame encodes a 233 amino acid long precursor protein with a signal peptide sequence for secretion and a consensus motif for lipid conjugation, which suggests that the mature protein is an exported lipoprotein antigen. The molecular mass of the mature protein antigen from M. leprae sonicate was shown to correspond to the deduced size of the M. tuberculosis protein by T cell Western analysis. Homology searches revealed two other similarly sized hypothetical secreted mycobacterial lipoproteins within the M. tuberculosis genome database. PMID- 9393627 TI - Release of cytokines and soluble cell surface molecules by PBMC after activation with the bispecific antibody CD3 x CD19. AB - Bispecific antibodies (BsAb) consist of two different heavy and light chains and may bind to two different antigens present on different cell types. With their dual specificity BsAb may recognize effector cells (e.g. T cells) on one hand and tumour cells (e.g. malignant B cells) on the other hand. The authors analysed whether T cell activation and subsequent killing of malignant B cells mediated by the bispecific antibody CD3 x CD19 was reflected by the release of cytokines. In addition, the authors investigated whether the in vitro cytokine release was similar to that observed in vivo in the patients treated with BsAb. The in vitro release of cytokines into the supernatant of cell cultures of peripheral blood mononuclear cells (PBMC) and malignant B cells was measured after incubation with either the bispecific antibody CD3 x CD19 or the monospecific anti-CD3 (aCD3) antibody in the presence or absence of interleukin (IL)-2. Release of tumour necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), IL-6, IL-8, IL 10, soluble (s) CD4, sCD8 and sCD25 by PBMC was equal under both conditions and could be used as an indicator for T cell activation. However, the cytokine pattern and level did not correlate with the cytotoxic capacity, which was 4 logs higher with BsAb + IL-2 compared to aCD3 + IL-2. The in vitro pattern of cytokine release was similar to that observed in vivo in the serum of patients treated with BsAb and IL-2, indicating the possibility of predicting cytokine release in future patients with other therapeutic regimens. PMID- 9393628 TI - Effect of treatments with cyclosporin A and anti-interferon-gamma antibodies on the mechanisms of immune tolerance in staphylococcal enterotoxin B primed mice. AB - The authors were interested to investigate the effect of Cyclosporin A (CsA), known to block interleukin-2 (IL-2) production, or of anti-interferon-gamma antibodies (anti-IFN-gamma Abs) in a model of T cell tolerance induced by the injection of the superantigen Staphylococcal Enterotoxin B (SEB) in BALB/c mice. After SEB immunization, tolerance was mainly achieved through deletion and anergy of SEB-reactive V beta 8+ T cells. Association of CsA treatment with SEB led to a greater decrease of the percentage of V beta 8+ CD4+ lymphocytes in the spleen and an abolition of clonal energy. In contrast, treatment of SEB primed mice with anti-IFN-gamma Abs resulted in an increased percentage of V beta 8+ CD4+ cells without affecting the induction of clonal anergy. The authors found that 1-2 h after SEB priming, splenic mRNA levels of IFN-gamma and IL-4 were decreased by either CsA and anti-IFN-gamma Abs, whereas FasL, Bcl-2, p. 53, and c-myc levels were not influenced by either treatment. However, SEB-induced IL-2 and IL-10 mRNA expression was suppressed only by CsA, whereas tumour necrosis factor-alpha (TNF alpha) was decreased only by anti-IFN-gamma Abs. To investigate whether the effect of CsA on the tolerance mechanisms was related to suppression of IL-2, CsA was administered together with recombinant IL-2. Whereas anergy was not influenced, the decreased percentage of V beta 8+ CD4+ cells seen in CsA-treated animals in the second week after SEB injection was partially corrected by the administration of IL-2. Experiments involving bromodeoxiuridine incorporation revealed that the latter effect of IL-2 was mainly due to a correction of the defective proliferation of V beta 8+ T cells after SEB injection in CsA-treated mice. These results suggest that the effect of CsA and anti-IFN-gamma Abs on tolerance mechanisms are in part explained by their action on cytokines. PMID- 9393629 TI - Increased mortality and impaired clonal deletion after staphylococcal enterotoxin B injection in old mice: relation to cytokines and nitric oxide production. AB - In the present study peripheral T cell tolerance and the occurrence of shock were evaluated in young and old mice after injection of Staphylococcal enterotoxin B (SEB). In young mice SEB immunization leads to tolerance based on deletion and anergy of SEB-reactive V beta 8+ T cells. With aging, mice developed resistance to SEB-induced deletion of V beta 8+ T cells as well as a high sensitivity to toxic shock. Compared to young mice, older mice injected with SEB showed increased serum levels of interferon-gamma (IFN-gamma), interleukin-2 (IL-2) and IL-4. These results were confirmed by reverse transcription-polymerase chain reaction (RT-PCR), as splenic mRNA levels taken 2 h after SEB injection showed higher values of IL-2, IL-4, and IFN-gamma in old mice. In contrast, mRNA levels for FasL and tumour necrosis factor-alpha (TNF-alpha) were lower. No difference in IL-10 mRNA was found. Compared to young mice, old mice showed a high, but statistically not significantly different (P = 0.20), production of nitric oxide (NO). Blocking of IFN-gamma with antibodies or reducing IFN-gamma by depletion of natural killer (NK) cells resulted, respectively, in a complete or partial protection against mortality in aged mice. Suppressing the NO production by the NO synthase inhibitor N-nitro-L-arginine methylester (L-NAME) increased the mortality in both young and old mice, and abrogated clonal deletion in the surviving young mice. In conclusion, in young mice NO production is a key factor in the protection against mortality and the development of clonal deletion after SEB injection. The higher mortality seen in older mice is mainly related to the elevated production of IFN-gamma and occurs despite a sufficient production of NO. The decreased clonal deletion of old mice may be related to their decreased expression of Fas ligand or TNF-alpha after SEB injection. PMID- 9393630 TI - Selective cytotoxicity of two rodent T cell lymphomas to rat yolk sac tumours involves a retroviral envelope protein expressed by the lymphoma. AB - A Gross virus induced rat T cell lymphoma G1-Tc1 and a Moloney virus induced mouse T cell lymphoma YAC-1 are shown to exert a strong cytotoxic activity against rat yolk sac tumours but not to various types of rat, mouse or human normal cells or tumour cell lines including carcinomas, sarcomas, lymphomas and gliomas. Both lymphomas are CD3+, CD4-, CD8- and T-cell receptor (TCR) alpha beta +. The cytotoxicity was not MHC restricted or dependent on the density of MHC class I of the target cells, and the mouse lymphoma killed the rat yolk sac tumour target. The cytotoxic action was fast and up to 80% specific killing was observed in 4-h 51Cr release assays. A rat B cell hybridoma was established from a Wistar/Furth (WF) rat immunized with the syngeneic lymphoma G1-Tc1 producing an immunoglobulin (Ig)G2c monoclonal antibody (MoAb) 1F2. This binds to the lymphomas G1-Tc1 and YAC-1 and also to a murine non-cytolytic Rauscher lymphoma RMA, but not to any other of several rat, mouse or human cell types tested. The 1F2 completely inhibited the killing of rat yolk sac tumours by the two cytolytic lymphomas, but did not interfere with the killing mediated by natural killer (NK) cells or cytolytic lymphokine-activated killer (LAK) cells. Immunochemical analysis of solubilized cell membranes of the lymphoma G1-Tc1 demonstrates that the 1F2 antibody recognizes an epitope on a retroviral gp 70 envelope protein. This indicates that a retroviral protein is involved in the lytic activity of the two lymphomas. PMID- 9393631 TI - Linomide enhances apoptosis in CD4+CD8+ thymocytes. AB - Linomide, a quinoline-3-carboxamide, has a pleiotropic immune modulating capacity and inhibits development as well as progression of disease in animal models of autoimmunity. Linomide treatment of mice resulted in a dramatic, dose-dependent decrease of the thymic cell number shortly after the start of administration. Flow cytometric analysis revealed that the major thymocyte subset, the early immature type CD4+CD8+ thymocytes, were reduced in number by 75%, mature CD4+CD8- or CD4-CD8+ thymocytes were less sensitive to treatment. The polyclonal T cell activator Con A (Concanavalin A) was used together with IL-2 to evaluate the potential proliferative responsiveness of ex vivo thymocytes. Thymocytes from mice treated with Linomide exhibited a more vigorous proliferation than control cultures. An effect shown to not only be due to the enrichment of mature thymocytes in the cultures from Linomide treated animals, but also when purified, mature thymocytes (CD4+CD8- and CD4-CD8+) were cultured with Con A and IL-2, these cells responded with a significantly enhanced proliferation. In vivo Linomide treatment did not result in increased plasma concentrations of corticosterone and treatment of adrenalectomized mice resulted in a reduction of thymocytes which was comparable to the effect in intact mice, indicating that glucocorticoids (GC) are not major mediators of Linomide-induced thymocyte deletion. In addition to this, and supporting a glucocorticoid independent mode of action, Linomide treatment of thymocytes in vitro resulted in a significant increase in the number of apoptotic cells, specifically in the CD4+CD8+ subset, implicating apopotosis as one component in the course of thymocyte reduction. In addition to this, in vivo treatment with Linomide resulted in an identical pattern to that seen in vitro in that there was significantly increased apoptosis only in the CD4+CD8+. These data indicate that Linomide modifies thymocyte development using a glucocorticoid independent pathway and results in the increased apoptosis of the CD4+CD8+ subset. PMID- 9393632 TI - Interferon-gamma production by human T cells and natural killer cells in vitro in response to antigens from the two intracellular pathogens Mycobacterium tuberculosis and Leishmania major. AB - Acquired resistance to both mycobacteria and Leishmania is primarily mediated by interferon-gamma (IFN-gamma), which triggers mechanisms leading to the death of the microorganism in macrophages. In this study, cell activation and IFN-gamma production was investigated in human peripheral blood mononuclear cells (PBMC) from individuals previously sensitized to tuberculin and without known exposure to Leishmania parasites. Immune staining for intracellular IFN-gamma and surface markers allowed flow cytometric identification of the cellular sources of IFN gamma in cell cultures incubated with purified protein derivative of tuberculin (PPD) and Leishmania antigens. It was found that IFN-gamma was produced in response to both PPD and Leishmania stimulant by T cells in the cultures. Activation of IFN-gamma producing natural killer (NK) cells was demonstrated only in some cultures, and only with concomitant T cell activation. PMID- 9393633 TI - Down-regulatory effect of Mycobacterium leprae cell wall lipids on phagocytosis, oxidative respiratory burst and tumour cell killing by mouse bone marrow derived macrophages. AB - The authors have previously demonstrated that lipids from Mycobacterium leprae cell walls inhibit macrophage functions and are endowed with anti-inflammatory properties in vivo. To investigate these observations further, the authors describe here the influence of dead M. leprae or of the lipids extracted from the cell wall of the mycobacterium, enclosed in liposomes, on the phagocytic, oxidative respiratory burst and tumouricidal ability of bone marrow derived macrophages in vitro. Dead M. leprae or its cell wall lipids abrogated the oxidative respiratory burst and phagocytic ability of mouse bone marrow derived macrophages. A dose-dependent inhibitory effect of the bacterial lipid extract on tumour cell killing by lipopolysaccharide (LPS)-activated bone marrow derived macrophages was demonstrated. However, when delipidated M. leprae was added to cultures of bone marrow derived macrophages, immune phagocytosis and superoxide production was up-regulated. Mycobacterium leprae or its lipids did not appear to be toxic to those cells assayed by the MTT (methyl thiazol tetrazolium) test. These data, added to our preceding observations, support the hypothesis that the down-regulatory activity of M. leprae wall lipids on macrophage function might be one of the evasive mechanisms of the bacterium to enable it to perpetuate itself in the host tissues. PMID- 9393634 TI - Nasal lymphoid tissue (NALT) as a mucosal immune inductive site. AB - Intranasal (i.n.) immunization is a very effective route for inducing mucosal immunity, but the cellular mechanism responsible for regulating and disseminating these responses is not fully understood. The authors studied the role of nasal lymphoid tissue (NALT) as a mucosal inductive site by using highly purified NALT cells obtained by a new method of mechanical isolation. The NALT cells, like Peyer's patch (PP) cells, were smaller in size and less granular than lymphocytes from salivary glands (SG) and small intestinal lamina propria (LP). The NALT cells isolated from i.n. immunized mice contained antigen-specific antibody secreting cells (ASC) predominantly of immunoglobulin (Ig)A isotype, similar to those also recovered from salivary glands in a time course study. However, the higher proportion of smaller sized sports formed by NALT cells in ELISPOT assays suggested that these cells were less differentiated precursors of those found in salivary glands. This was supported by the fact that after i.n. immunization, IgA ASC appeared in NALT, as well as in mucosal effector sites SG and LP, but none or very few in another mucosal inductive site, PP. In contrast, after intragastric (i.g.) immunization, IgA ASC were detected in PP, along with the SG and LP, but none or very few in NALT. Furthermore, after i.n. immunization, lymphocytes from NALT but not PP proliferated in response to the specific antigen in culture. These findings imply that NALT served as an inductive site for IgA antibody responses at mucosal effector sites. PMID- 9393635 TI - How to diagnose chronic rejection. A study in porcine intestinal allografts. AB - Porcine small bowel allografts were followed for 18 weeks during immunosuppression with cyclosporine-A (CyA), azatioprine and prednisone. The mucosal alterations noted at the 12th week were epithelial vacuolation and loss of Goblet cells. Moderate infiltration of inflammatory cells, mainly lymphocytes, was found in the lamina propria. In addition, a few grafts exhibited oedema and fibrosis. Vessels already showed endothelial swelling and intimal proliferation at the 12th week. In the submucosa, the infiltration of inflammatory cells was not present till the 18th week. Further changes in the mucosa at the 18th week were the blunting of villi, cuboidal epithelium, crypt abscesses and epithelial atrophy. The histological alterations of mucosa and lamina propria existing in the full thickness biopsies were mostly also detectable in mucosal biopsies, provided that multiple biopsies were taken. Thus these parameters analyzed from mucosal biopsy material are suitable for the diagnosis and monitoring of chronic small bowel rejection. In autopsy, the most prominent features were in the mesenterial arteries: intimal proliferation, vasculitis, proliferation of media and endothelial alterations. The activity of the mucosal disaccharidases maltase and sucrase remained near the initial level till the 12th week and had decreased markedly by the 18th week. PMID- 9393636 TI - T-cell receptor BJ gene segment expression in human umbilical cord blood CD4+ and CD8+ T-cell subsets. AB - By employing RT-PCR-based technology, followed by Southern-blot analysis, patterns of relative TRC BJ gene segment usage in human CD4+ and CD8+ umbilical cord blood T cells (UCT) from ten children were determined in relation to seven recombined TCR BV gene (sub) families (BV 3, 5S1, 6S1-3, 8, 9, 12 and 18). Normal frequency of usage of individual BJ members was observed to be extremely nonrandom. BJ usage in association with each BV was ranked and mean ranking values were calculated for individual BJs. Moreover, BJ family usage and family ranges as well as individual BJ over-representations were determined. In all these aspects of BJ exon expression, CD4+ and CD8+ UCT displayed similar distribution patterns. Comparisons of BJ usage in UCT subpopulations and in the adult peripheral blood lymphocyte (PBL) counterparts were performed and many similarities were observed. However, discrepancies in two parameters were recorded; contrary to observations in PBL, individual BJ over-representations were virtually absent in UCT, and significantly less wide BJ family ranges were demonstrated in CD8+ UCT relative to CD8+ PBL T cells. These differences support the notion that UCT are in a less dynamic state than are PBL T cells. Hence, despite the fact that PBL T cells are subjected to continuous antigenic challenge, the striking resemblance of PBL and UCT with regard to the overall individual relative usage, ranking, mean ranking and family utilisation of BJ gene segments, irrespective of the choice of recombined BV exons, may suggest a relatively nondiscriminatory role for the BJ gene product in antigen recognition as compared to those encoded by the BV, (N) and BD gene segments. PMID- 9393637 TI - Cloning and molecular analyses of the Arabidopsis thaliana plastid pyruvate dehydrogenase subunits. AB - Herein we report the first molecular description of the pyruvate dehydrogenase component of the higher plant plastid pyruvate dehydrogenase complex. The full length cDNAs for the E1 alpha (1530 bp) and E1 beta (1441 bp) subunits of the Arabidopsis thaliana plastid pyruvate dehydrogenase contain open reading frames that encode polypeptides of 428 and 406 amino acids, respectively, with calculated molecular weight values of 47,120 and 44,208. The deduced amino acid sequences for Arabidopsis plastid E1 alpha and E1 beta have 61% and 68% identity to the odpA and odpB genes of the red alga Porphyra purpurea, respectively, but only 31% and 32% identity to the plant mitochondrial counterparts. Results of Southern analyses suggest that each subunit is encoded by a single gene. Northern blot analyses indicate expression of mRNAs of the appropriate size in Arabidopsis leaves. PMID- 9393638 TI - Patch clamp investigation into the phosphate carrier from Saccharomyces cerevisiae mitochondria. AB - After heterologous expression in E. coli, functionally active phosphate carrier (PIC) from Saccharomyces cerevisiae mitochondria was purified and reconstituted into giant liposomes and used for patch clamp experiments. Single channel currents across excised patches revealed an anion channel function of the PIC protein. Besides the three transport modes known to date, namely phosphate/phosphate exchange, phosphate/OH exchange and mercurial-induced unidirectional transport, this channel activity represents the fourth transport mode of the PIC. The PIC channel activity was sensitive towards phosphate as its physiological substrate. Phosphate (10 mM) blocked in a specific but reversible manner the PIC channel, suggesting a phosphate-dependent conformational change of the protein into the carrier mode. Furthermore, the current through the channel and its gating activity were affected by divalent cations. In the presence of Ca2+ and Mg2+, the channel displayed a mean conductance of 25 +/- 5 pS whereas 40 +/- 10 pS was observed in the absence of divalent cations. Also, the dwell times in either the open or closed state of the PIC channel appeared to be prolonged in the presence of Ca2+ and Mg2+. The observed PIC channel characteristics are discussed with respect to previously reported electrophysiological in situ measurements on anion channels of the inner mitochondrial membrane. Similarities of the PIC channel to the inner mitochondrial anion channel (IMAC) have been found. PMID- 9393639 TI - Intramolecular oxidation of cytochrome c by covalently attached sulfoaromatic molecules. AB - Two photosensitive molecules, 1-maleimidopyrene-3,6,8-trisulfonate (MPTS) and N acetylaminoethyl-1-aminonaphthalene-5-sulfonate (AEDANS), are employed to drive the intramolecular oxidation of the heme residue in cytochrome c. Intense pulse illumination (60-120 MW cm-2) of MPTS and AEDANS in the aqueous solution by the third harmonic frequency of Nd-Yag laser drives a two successive-photon process of the dyes. The oxidized products originating from the dyes react with variety of electron donors. MPTS and AEDANS residues were covalently linked the Saccharomyces cerevisiae iso-1-cytochrome c by labeling of its single sulfhydryl group. When pulsed by intensive laser beam the heme of the labeled ferrocytochrome c undergoes fast oxidation. Transient absorption spectroscopy was used to directly measure the rate constants for the photoinduced electron transfer reaction from the ferros heme group to the oxidized dyes. The rate constant was found to be (3.6 +/- 0.4) x 10(4) s-1 for MPTS derivative. The rate of the heme oxidation in AEDANS derivative was faster than response time of our detection system (20 ns). Rapid photooxidation of cytochrome c makes it a useful tool for fast initiation of electron transfer in oxidized direction within complexes of cytochrome c with the other redox proteins. PMID- 9393640 TI - The role of the membrane-spanning and extra-membranous regions of the iron-sulfur protein in its assembly into the cytochrome bc1 complex of yeast mitochondria. AB - The assembly of six deletion mutants of the Rieske iron-sulfur protein into the cytochrome bc1 complex was investigated by immunoprecipitation from detergent solubilized mitochondria with specific antisera against either the iron-sulfur protein or the intact cytochrome bc1 complex. After import, the mutant proteins lacking residues 41-55 or 66-78, located at the membrane-spanning region of the protein, and residues 182-196 located at the C-terminus of the protein, were assembled in vitro into the bc1 complex approximately 50% as effectively as the wild type iron-sulfur protein suggesting that these regions of the iron-sulfur protein may not be critical for the assembly. By contrast, only trace amounts of the mutant proteins lacking residues 80-95, 122-135, 138-153 located in the extra membranous region of the iron-sulfur protein were assembled into the bc1 complex. After import in vitro into mitochondria isolated from a cytochrome b-deficient yeast strain, the mutants lacking residues 41-55 and 182-196 were assembled as efficiently as the wild type; however, the mutants lacking residues 55-66 and 66 78 were assembled less efficiently in the absence of cytochrome b suggesting that the hydrophobic membrane-spanning region, residues 55-78, of the iron-sulfur protein, may interact with cytochrome b during the assembly of the bc1 complex. PMID- 9393641 TI - Binding of MOA-stilbene to the mitochondrial cytochrome bc1 complex is affected by the protonation state of a redox-Bohr group of the 'Rieske' iron-sulfur protein. AB - MOA-stilbene is a specific inhibitor of the ubihydroquinone oxidation center (center P or o) of cytochrome bc1 complex. Binding of this inhibitor does not require the 'Rieske' iron-sulfur protein, but is affected by the redox-state of the cytochrome bc1 complex. We have analyzed the pH dependence of the apparent dissociation constant for MOA-stilbene. A 2.5 fold change in affinity between pH 6.0 and 9.5 was observed for oxidized bovine cytochrome bc1 complex. The pH profile could be simulated by assuming a single protonable group with pKA = 7.7. This pKA was not observed after partial or complete reduction of the enzyme or after removal of the iron-sulfur protein. We conclude that this protonable group was identical to the redox-Bohr group with the same pKA that has been reported to be associated with the 'Rieske' iron-sulfur cluster. Fully reduced cytochrome bc1 complex exhibited an additional binding site for MOA-stilbene. As this second binding site was abolished by the center P inhibitor stigmatellin, but not by antimycin, an inhibitor of ubiquinone reduction at center N, we conclude that it is also located at center P. PMID- 9393642 TI - Effect of nitroso compounds on Na/K-ATPase. AB - Thiol containing NO.-derivatives were found to inhibit the activity of brain and kidney Na/K-ATPase. S-Nitrosogluthatione demonstrated only minor inhibiting activity, while dinitrosyl iron complexes (DNIC) with cysteine or glutathione were much more effective. Brain Na/K-ATPase is more vulnerable to inhibiting action than kidney Na/K-ATPase. Inhibition of the activity is accompanied by a decrease in amount of protein thiol groups and a change in the substrate dependence curve of the enzyme. Restoration of Na/K-ATPase activity by SH-reagent dithiothreitol or cysteine is accompanied by restoration of SH-groups of the enzyme. This suggests that blockade of SH-groups of Na/K-ATPase is responsible for the inhibition. The possibility that this blockade results in disordering of interprotomer interactions within the oligomeric complexes of Na/K-ATPase is suggested. Possible regulatory meaning of the effect of NO. derivatives is discussed. PMID- 9393643 TI - ATPase and phosphatase activities are differentially inhibited by photo-oxidation of the sarcoplasmic reticulum Ca(2+)-ATPase. AB - We have already described that photo-oxidation of the sarcoplasmic reticulum Ca(2+)-ATPase with the halogenated dye erythrosin B produces inhibition of the ATPase activity (J.A. Mignaco et al., Biochemistry 35 (1996) 3886-3891). We now show that the Ca(2+)-dependent and Ca(2+)-independent p-nitrophenylphosphatase activities are also inhibited by this treatment. Modification of rapidly (< 10 min) oxidized residue(s) is responsible for the major loss of ATPase activity, whereas photo-inhibition of the phosphatase activities occurs more slowly (t1/2 20-30 min). Here we have focused on photo-inhibition of the Ca(2+)-independent pNPPase activity, and the counteracting effects of ATP and FITC. Following photo oxidation, the Ca(2+)-independent pNPPase activity decreases monotonically. ATP partially protects against the inactivation of the pNPPase, whereas labeling the enzyme with FITC does not. However, the protective effect of ATP is completely abolished by the attached FITC. These data are interpreted in terms of two different sites that are susceptible to photo-oxidation and are involved in different events related to substrate hydrolysis. PMID- 9393644 TI - Microbiological observations in the anoxic basin Golfo Dulce, Costa Rica. AB - Our basic microbiological studies of the water column and the sediment of Golfo Dulce, Costa Rica, were focused on aerobic and denitrifying sulfur-oxidizing bacteria and anaerobic sulfate-reducing bacteria. We observed no increasing numbers of total bacterial counts within the water column. Although no oxygen was present hydrogen sulfide was only detectable close to the sediment. The highest numbers of sulfate-reducing bacteria measured by Most-Probable-Number counts were found in or close to the sediment. In the anoxic bottom water sulfide-oxidizing bacteria typically containing large sulfur globules were observed microscopically. They were identified as free-swimming Thiovulum and Thiospira species. At one station large vacuolated forms of the filamentous colourless sulfur bacterium Beggiatoa were noted. Together with these sulfur containing bacteria there were long free swimming rods showing no sulfur inclusions of unknown character. The microscopic observations showed good correlation with Most Probable-Number-counts and molecular biological techniques for sulfate-reducing bacteria. PMID- 9393645 TI - Polychaete worms (Annelida) collected in Golfo Dulce, during the Victor Hensen Costa Rica expedition (1993/1994). AB - A total of forty seven species of benthic polychaetes belonging to twenty five families have been identified from bottom samples taken in Golfo Dulce, Costa Rica, by the RV Victor Hensen. Only those stations collected in waters less than 100 m depth contained polychaetes. The major feeding type of these polychaete species was surface deposit feeding with slightly fewer species recognized as carnivores. Comparison of the species identified from the RV Victor Hensen material with those of the RV. T.V. Thompson material collected in 1969 reveals very few that are common to the two studies, indicating that the polychaetes of Golfo Dulce are probably quite diverse and poorly known. An estimated total of eighty five species of polychaetes have been identified from Golfo Dulce in the two studies. PMID- 9393646 TI - Annotated list of species of marine crustaceans (Decapoda and Stomatopoda) from Golfo Dulce, Costa Rica. AB - The present study is an annotated list of the marine crustaceans (Decapoda and Stomatopoda) from Golfo Dulce, Costa Rica, collected during the RV Victor Hensen Cruise 1993-1994, at depths ranging from 20 to 260 m. Stomatopoda was represented by one family and two species. Decapoda was represented by 12 families and 21 species. Two new records for Costa Rica, Cancer johngarthi and Lysmata californica are reported. This increases the reported marine crustaceans of Golfo Dulce to three species of Stomatopoda and sixty-six of Decapoda (a complete checklist in included). A list of species reported in the literature from the gulf is also included. PMID- 9393647 TI - Checklist of crustaceans (Decapoda and Stomatopoda), collected during the Victor Hensen Costa Rica expedition (1993/1994). AB - A checklist of the marine crustaceans (Decapoda and Stomatopoda) from the Pacific coast of Costa Rica collected by trawling during the RV Victor Hensen Cruise (1993-1994) is presented. A total of 117 species were identified, 10 stomatopods and 107 decapods. PMID- 9393648 TI - Checklist of copepods from Gulf of Nicoya, Coronado Bay and Golfo Dulce, Pacific coast of Costa Rica, with comments on their distribution. AB - A list of 54 copepod species (Crustacea) in 23 families is presented for the Pacific coast of Costa Rica. Identifications are from zooplankton samples of the Victor Hensen Expedition during December 1993 and February 1994. Samples were taken with a Bongo net (0.60 m net opening, 2.50 m net length) with 200 microns mesh size. Oblique hauls were done from the surface to the ground at a towing speed of aprox. 1 knot. 37 species (68.5%) were found in the Gulf of Nicoya, 36 in Golfo Dulce (66.6%) and 17 (31.4%) species were common to both gulfs, while only twelve species (22.2%) were found in Coronado Bay. Four species (7.4%) were distributed along the coast and were common to the three regions: Paracalanus parvus, Euchaeta sp., Oithona plumifera and O. similis. Eleven species of calanoids found normally in the Costa Rica Dome show the influence of typical oceanic waters principally at the mouth of Gulf of Nicoya. Differences were observed in the composition and presence of the copepod species when the inner and outer (upper and lower) parts of both gulfs were compared. Gulf of Nicoya was dominated in its upper part by typical neritic estuarine species like Acartia lilljenborgii, Paracalanus parvus and, Hemyciclops thalassius as well as species of Pseudodiaptomus. On the other hand a more oceanic composition of copepods was observed in the lower part of the gulf. Both small species, like Oncaea venusta, as well as larger species, such as Pleuromamma robusta, Eucalanus attenuatus, E. elongatus and Rhincalanus nasutus, were typical of these waters. Oithona plumifera and O. similis were found in the lower part too; and both species are typical from oceanic water. Coronado Bay was characterized by the presence of typical oceanic species like Neocalanus gracilis, Euchaeta longicornis, Eucalanus attenuatus and Haloptilus ornatus with more transitional species like Clausocalanus pergens and C. furcatus near the coast. In the Golfo Dulce differences in copepod composition were also observed, but the separation of the species was not so evident. Outer stations were represented by oceanic species like Paracalanus aculeatus, Pleuromamma gracilis, Lucicutia ovalis, Candacia catula, Euchaeta wolfendeni and Oncaea mediterranea, while the inner station, located at the upper part of the Gulf, was more characterized by a mixed copepod group, with both neritic species like Pseudodiaptomus wrigthi, Acartia danae, A. clausi, Canthocalanus pauper as well as oceanic species like Scolicithricella marginata, Saphirina nicromaculata or Oncaea conifera. Two species of Coryceaus, C. flaccus and C. speciosus, were identified in the outer stations of Golfo Dulce, while C. brehmi was found in inner stations of Gulf of Nicoya. The majority of copepods found are typical of the east Pacific. This paper constitutes an additional work about the copepods in the Gulf of Nicoya and the first report of copepod species for Coronado Bay and Golfo Dulce. PMID- 9393649 TI - Demersal crustacean assemblages along the Pacific coast of Costa Rica: a quantitative and multivariate assessment based on the Victor Hensen Costa Rica expedition (1993/1994). AB - During the first cruise leg with the RV Victor Hensen to the Pacific coast of Costa Rica in December 1993 (end of the rainy season) the crustacean fauna found in the demersal collections revealed an unexpected species richness and biomass. The Crustacea collections were analyzed qualitatively and quantitatively during the fourth leg (February 1994, dry season) in the three study areas Golfo Dulce (GD), Bahia Coronado in the Sierpe-Terraba-estuary (ST) and Golfo de Nicoya (GN). Qualitative data were available for comparison from the first leg in december 1993. A total of 24 beamtrawl and ten ottertrawl sample collections were done on an area of 860.000 m2 yielding a total of 119 species with a biomass of 37.8 kg (10275 specimens). Despite the smaller area covered by the beamtrawl, it collected a higher number of species and more biomass than the ottertrawl due to the smaller mesh size (0.8 cm). Judging from the shape of the species -per-area curves, the crustacean fauna appeared as representatively sampled for the study area. As compared with the GN (biomass 0.36 g +/- 0.26, SR = 97) and the ST (0.41 g +/- 0.27, SR = 59) and according to the results of the log-series-plots constructed from the abundance data, the GD seems to be a depauperated area with significantly lower biomass (0.05 g +/- 0.07) and species richness (45 sp.). No crustaceans were found in the center of the deep basin of the GD put parts of the interior gulf with adjacent mangrove areas seem to be important as nursery area for some commercially important penaeid shrimp species. The ST-estuary revealed the highest mean species number per station in the whole study area, but the GN had the highest total number of species. Biomass seems to be regularly distributed and not depth-depending within the GN, while species abundance varies clearly, confirming previous results. In contrast, abundance and biomass correlated well in the ST. Based on the results of the multivariate analysis, seven station groups of particular species assemblages can be distinguished in the study areas. Despite a high variability between stations in abundance and biomass, the following four areas of characteristic species assemblages can be identified which are also confirmed by an independent study on demersal fishes: (1) the interior part of the GN, characterized by juvenile shrimps (Sicyonia disdorsalis, Trachypenaeus fuscina), several patchily distributed anomurans, brachyurans and other predator species like portunids (especially Portunus asper) and pre-adult stomatopods (Squilla spp.); (2) the exterior part of the GN with high amounts of caridean (Pantomus affins, Plesionika spp.) and penaeid shrimps (Sicyonia picta, Solenocera mutator), the highly abundant Iliacantha hancocki and some specimens of the stomatopod Hemisquilla stylifera and the deep water portunid Portunus iridescens; (3) a transition zone between 60 and 120 m water depth with a heterogeneous faunal composition, located in the ST and east of Isla Tortugas in the GN, and (4) the oxygen-depleted shelf edge area, dominated by the galatheid Pleuroncodes monodon. Mass occurrence of this species takes place off the GD and to a lesser extent off the ST-estuary, associated with high numbers of Solenocera spp. There seems to be a general trend of species groupings along abiotic gradients (depth, temperature, oxygen saturation) interrupted by small scale variations in habitat type, current regime, food availability and other factors not identified in this study. Neither total abundance and biomass nor biotic summary parameters like diversity, dominance or species richness correlated well with the abiotic factors measured during this survey. PMID- 9393650 TI - Comparative biomass spectra and species composition of the zooplankton communities in Golfo Dulce and Golfo de Nicoya, Pacific coast of Costa Rica. AB - This study is based on a subset of plankton samples obtained during an expedition of the German RV Victor Hensen to the Pacific coast of Costa Rica in 1993/94. It aims at the identification of the main plankton taxa for a general description and comparison of the plankton communities of the gulf systems Golfo de Nicoya (GN) and Golfo Dulce (GD) and the analysis of biomass spectra at inshore and offshore stations at the end of the rainy season and during the dry season. Inshore plankton biomass was significantly higher in GN than GD and exceeded offshore biomass several times, while in the GD area the reverse was found. In the rainy season, inshore biomass spectra of GN and GD were discontinuous with biomass concentrations at small sizes (around 0.06 mg) suggesting little developed communities, with highest production and energy use occurring in the small organisms. From the rainy to the dry season inshore species richness increased in both gulf systems and a shift was observed towards the larger size groups resulting in more continuous biomass spectra. In GN, bivalve larvae, foraminifers, ostracods, mysids and nauplii increase heavily in abundance and some gelatinous specimens occur. In GD, gelatinous zooplankton appears in enormous abundance and dominate the community biomass, followed by large chaetognaths and ostracods. In GD, inshore plankton has neritic and oceanic elements and differs less from the offshore plankton, whereas in GN, inshore plankton in largely neritic. The high abundance of fish eggs and invertebrate larvae suggest that this area is an important spawning ground. While in the rainy season inshore biomass was about 15 times higher in GN compared to GD, this difference was reduced to 3-4 times in the dry season due to the appearance of the large predators mentioned above. The changes from the rainy to the dry season at the offshore stations of both gulf systems are less pronounced in terms of total biomass, shape of the biomass spectra and taxonomic composition of the community. The differences-relatively continuous biomass spectra with an increasing slope and a high total biomass in GD versus flat and shorter spectra due to the absence of large chaetognaths and medusa in the GN-suggest that conditions in the former area allow for a better development of a trophodinamically tightly structured plankton community. PMID- 9393651 TI - Distribution and biomass of arrow worms (Chaetognatha) in Golfo de Nicoya and Golfo Dulce, Costa Rica. AB - The chaetognath species guild was analyzed from samples collected during the cruise of the German RV Victor Hensen to the Pacific coast of Costa Rica in December 1993 and February, 1994, finding the following ten species of the genera Sagitta and Krohnitta: S. enflata, S. hexaptera, S. pacifica, S. neglecta, S. regularis, S. bedoti, S. friderici, S. popovicii, S. pulchra and K. pacifica. Because of their distributional patterns in the study area these species were ascribed to the following ecological groups: neritic, semi-neritic and oceanic. A strong gradient in species richness from offshore to inshore waters (8 to one respectively) was found in both gulf systems. Inshore chaetognaths were dominated by juveniles and adults of S. friderici in Golfo de Nicoya and by S. popovicii in Golfo Dulce. Biomass spectra were more continuous and of wider range in the Golfo Dulce area showing a dominance of larger chaetognaths, suggesting a more general developed pelagic system in Golfo Dulce, where larger chaetognaths might structure the plankton community by strong grazing pressure from above. PMID- 9393652 TI - Ichthyoplankton assemblages in the Gulf of Nicoya and Golfo Dulce embayments, Pacific coast of Costa Rica. AB - Ichthyoplankton surveys were conducted in December (rainy season), 1993 and February (dry season), 1994, during the RV Victor Hensen German-Costa Rican Expedition to the Gulf of Nicoya and Gulfo Dulce, Costa Rica. Samples from the inner, central, and outer areas of each gulf were collected in oblique tows with a bongo net of 0.6 m mouth diameter, 2.5 m long and 1000-micron mesh. A total of 416 fish larvae of 22 families were sorted out of 14 samples. Stations of both the maximum (11) and the minimum (1) family richness were located in Golfo Dulce. Mean total larval abundances were 124.9 and 197.2 individuals 10 m-2 for the Gulf of Nicoya and Golfo Dulce, respectively, while mean larval densities ranged from 95.3 larvae 10 m-2 in December to 236.7 larvae 10 m-2 in February. However, no statistical differences between gulfs or seasons were detected, due to the high within-group variability. Cluster Analysis, Multi-Dimensional Scaling (MDS), and non-parametric tests showed two well-defined major groups: (1) the Gulf of Nicoya neritic assemblage, represented by Engraulids, Sciaenids, and Gobiids (inner and central stations), and (2) the oceanic assemblage, dominated by Myctophids, Bregmacerotids, Ophiidids, and Trichiurids (outer stations off the Gulf of Nicoya and Golfo Dulce). A third, although less defined group, was an Ophichthid dominated assemblage (typical in areas nearby coral or rocky reefs). These assemblages closely resemble the clusters based upon adult fish data of the beamtrawl catches of the same cruise. This publication is the first to report on the ichthyoplankton community of Golfo Dulce. PMID- 9393653 TI - Fishes collected during the Victor Hensen Costa Rica expedition (1993/1994). AB - A list is presented of 242 species of fishes taken in the Golfo de Nicoya, Golfo Dulce and on the Pacific continental shelf of Costa Rica. The specimens were collected using dredges and bottom trawls during December 1993 and February 1994. The Golfo Dulce revealed the lowest diversity with only 75 species represented; 118 species were collected in the Golfo de Nicoya and 129 species in offshore waters. It is presumed that low fish diversity in Golfo Dulce is due to the deep, unproductive waters in that embayment. The checklist includes presence-absence data for each locality. PMID- 9393654 TI - Demersal fish assemblages along the Pacific coast of Costa Rica: a quantitative and multivariate assessment based on the Victor Hensen Costa Rica expedition (1993/1994). AB - During two cruise legs with the RV Victor Hensen (December 1993, February 1994), the demersal fish assemblages of the Golfo de Nicoya (GN), Bahia Coronado-Sierpe Terraba (ST) and Golfo Dulce (GD) areas were assessed from nearshore (approximately 20 m) to shelf edge (approximately 200 m) waters. 44 Beam- and 29 otter trawl collections were made on an area of 2,119,405 m2, yielding a total of 242 species of fish. Despite the lower number of samples taken, more species were collected by the otter trawl (189 compared to 160), due to a wider area swept. As revealed by the species-area curve and a longnormal-curve constructed from the pooled (log) abundance data, the fish assemblage appeared as well sampled and a theoretical species richness (SR) of-306 was estimated for the whole area. Mean species number per collection and mean biomass per area were much lower in the GD area (9.3 species, 0.36 g/m2) compared to the ST (15.4, 0.81 g/m2) and GN (17.3, 0.74 g/m2) areas, indicating a depauperate fish assemblage in the former. Lowest species numbers and biomass were found in the central deep part of GD with increasing values towards the sill area at the opening of the gulf and towards the shallow stations above the thermocline. Average biomass was an order of magnitude higher in the interior part of GN compared to the other areas with values up to 18.1 g/m2. Based on results of a multivariate analysis of the collections, the GN area can be divided into (1) an interior shallow area above the thermocline (< 50 m) characterized by scianids, sea carfishes, stingrays, flatfishes, sea robins, (2) an outer part (> 100 m) characterized by cods, scorpionfishes, gobies, cutlassfishes, serranids, anglerfishes and flatfishes and (3) a transition zone of the central and lateral parts with a mixed species assemblage with carangids, pufferfish, snappers, several flatfish species and the lizardfish as common elements. Characteristic for the deep basin of GD were small species of the genera Cynoscion and Porichthys. These occurred in low densities, suggesting a reduced carrying capacity of this deep basin for fish biomass in terms of food and oxygen. Species occurring at the shallow stations of GD are also found at a similar depth in the other areas, but many species are missing, namely ariids and many scianids found in the GN area. The species assemblage of the ST area resembles that of GN. Ariids, however, are missing here too. Biotic station parameters like species richness, biomass, abundance and production were not significantly correlated with abiotic parameters (temperature, oxygen, nutrients) suggesting that other habitat factors not evaluated in this study like habitat heterogeneity, distance to the open ocean, current regime and food availability probably are important factors for the structure of the fish assemblage. PMID- 9393655 TI - D-dimer levels detected DVT in patients hospitalized for stroke rehabilitation. PMID- 9393656 TI - Viral gastroenteritis associated with eating oysters -- Louisiana, December 1996 January 1997. AB - Viral gastroenteritis outbreaks caused by caliciviruses (i.e., Norwalk-like viruses or small round-structured viruses) have been associated with eating contaminated shellfish, particularly oysters (Crassostrea virginica). This report describes the findings of the investigation of an outbreak of oyster-associated viral gastroenteritis in Louisiana during the 1996-97 winter season and implicates sewage from oyster harvesting vessels as the probable cause of contaminated oysters. PMID- 9393657 TI - Progress toward poliomyelitis eradication -- Western Pacific Region, January 1, 1996-September 27, 1997. AB - In 1988, the World Health Assembly adopted the goal of global poliomyelitis eradication by 2000, which was endorsed in each of the six regions of the World Health Organization (WHO). In the Western Pacific Region (WPR), where the last known case of polio associated with isolation of wild poliovirus occurred in March 1997, the reported number of cases decreased from 5963 in 1990 to 197 in 1996. This report documents progress toward polio eradication in WPR from January 1, 1996, through September 27, 1997, in countries where polio is endemic (Cambodia, China, Laos, Papua New Guinea, Philippines, and Vietnam) or recently was endemic (Malaysia and Mongolia) and describes the routine and supplemental vaccination activities necessary to interrupt wild poliovirus transmission in the region. PMID- 9393658 TI - Surveillance for fetal alcohol syndrome using multiple sources -- Atlanta, Georgia, 1981-1989. AB - Fetal alcohol syndrome (FAS) is caused by heavy alcohol consumption during pregnancy and is characterized by specific anomalies of the face; prenatal and postnatal growth deficits; and a variety of central nervous system (CNS) abnormalities, including mental retardation. Children with either full or partial FAS often incur severe and costly secondary disabilities. Despite the importance of surveillance for establishing the magnitude of FAS and in monitoring trends in the occurrence of this disease, population-based surveillance for FAS has been difficult because the syndrome can be diagnosed only by clinical observation and often is not recognized until after the child reaches school age. Although most FAS surveillance has been based on diagnoses among newborns, most (89%) cases (full FAS and partial FAS) are diagnosed after the age of 6 years. To develop a more accurate estimate of the prevalence of FAS in a defined population, in 1997 CDC linked data from the Metropolitan Atlanta Congenital Defects Program (MACDP) and the Metropolitan Atlanta Developmental Disabilities Surveillance Program (MADDSP) for children born in Atlanta during 1981-1989 (the most recent birth year for which data were available for 3-10-year-olds). This report presents a multiple-source method for FAS surveillance that is more complete than previous methods and that enables comparison of rates between states. PMID- 9393659 TI - Youth Risk Behavior Surveillance: National College Health Risk Behavior Survey- United States, 1995. AB - PROBLEM/CONDITION: Colleges and universities are important settings for delivering health promotion education and services to many young adults. However, before this national college-based survey was conducted in 1995, the prevalences of health-risk behaviors among college students nationwide had not been well characterized. REPORTING PERIOD: January through June 1995. DESCRIPTION OF THE SYSTEM: The Youth Risk Behavior Surveillance System (YRBSS) monitors six categories of priority health-risk behaviors among youth and young adults: behaviors that contribute to unintentional and intentional injuries, tobacco use, alcohol and other drug use, sexual behaviors, unhealthy dietary behaviors, and physical inactivity. The YRBSS includes a) national, state, and local school based surveys of high school students conducted biennially since 1991, b) a household-based survey conducted in 1992 among a national sample of youth aged 12 21 years, whether enrolled in school, and c) the national college-based survey conducted in 1995. This report summarizes results from the national college-based survey-the National College Health Risk Behavior Survey (NCHRBS)-and describes priority health-risk behaviors among college students nationwide and health promotion programs on college campuses. RESULTS AND INTERPRETATION: Data from the 1995 survey indicated that many college students throughout the United States engage in behaviors that place them at risk for serious health problems. Almost one third (29.0%) of college students were current cigarette smokers. One third (34.5%) of college students reported episodic heavy drinking during the 30 days preceding the survey, 27.4% reported drinking alcohol and driving during the 30 days preceding the survey, and 30.5% of students who had gone boating or swimming during the 12 months preceding the survey had drunk alcohol while boating or swimming. One in five (20.4%) female college students had been forced to have sexual intercourse during her lifetime. Only 29.6% of students who had had sexual intercourse during the 3 months preceding the survey had used a condom at last sexual intercourse, and 34.5% had used birth control pills. Approximately one in five (20.5%) college students was overweight. Survey results indicated that three fourths (73.7%) of students had failed to eat five or more servings of fruits and vegetables on the day preceding the survey, 21.8% had eaten three or more high fat foods on the day preceding the survey, and few students had engaged in vigorous (37.6%) or moderate (19.5%) physical activity at recommended levels. ACTIONS TAKEN: The NCHRBS data will be used to measure progress toward achieving 28 national health objectives related to the health-risk behaviors of college students and two national health objectives related to the availability and characteristics of health promotion programs for college students. These data also will be used nationwide by college health and education officials to improve health policies and programs designed to reduce risks associated with the leading causes of mortality and morbidity among college students. PMID- 9393660 TI - Secondary ischemic tolerance improved by administration of L-NAME in rat flaps. AB - Nitric oxide (NO) under basal conditions is an important regulator of vascular tone. Under ischemic conditions, however, NO can combine with superoxide anion to produce the damaging hydroxyl free radical. The current project observes the effect of inhibiting NO production (L-Nitro-amino-methyl-arginine, L-NAME) on flaps rendered ischemic by secondary (2 degrees) venous obstruction. Eighty rats had 3 x 6 cm skin flaps based on the epigastric vessels. Primary (1 degree) ischemia was produced by arteriovenous occlusion for 2 hours; (2 degrees) venous ischemia was induced by clamping the vein, alone for either 3 or 5 hours. Thirty minutes prior to 2 degrees ischemia, rats received either L-NAME (30 mg/kg) or saline buffer. Flap survival was assessed 7 days later and Chi-square analysis was used. At 3 hours of ischemia, treatment improved survival from 55% to 85% (P < 0.05). Treatment also improved survival at 5 hours of ischemia from 5% to 35% (P < 0.04). Although under resting conditions, NO is a potent vasodilator, during 2 degrees venous obstruction it may contribute to flap necrosis. PMID- 9393661 TI - A new method of applying fibrin glue at the microvascular anastomotic site: the "paintbrush" technique. AB - Fibrin glue has been used by several researchers to seal microvascular anastomoses sites. Usually the glue is delivered at the site with a syringe and needle. However, this technique can result in excess deposition of glue, which can cause intravascular thrombosis and can harden the vessel wall. We describe a new technique that avoids both these problems. We designed a "paintbrush" by inserting a suture through the syringe needle and then using the suture tip as a brush to apply the fibrin glue. End-to-side anastomoses were performed between the femoral artery and femoral vein in ten rats. Only two sutures were used to perform the anastomosis, the rest of the anastomosis being completed by sealing the gaps with fibrin glue. All the anastomoses were patent after 2 weeks, and no thromboses occurred. This is a new method for applying fibrin glue that delivers an adequate amount of fibrin glue without causing thrombosis. PMID- 9393662 TI - Results of interfascicular nerve grafting for radial nerve lesions. AB - During the 10 year interval 1979-1989, 20 patients underwent nerve grafting of a radial nerve lesion, 13 high radial and 7 posterior interosseous. Average follow up was 38 months (range 12 months-10 years). Overall 72% of patients achieved a Highet Scale rating of M3 or better function and 44% M4 or better recovery. Age of the patient and length of the nerve graft did not seem to influence outcome. Time from initial injury to nerve grafting did affect outcome, with 85% of patients grafted within 6 months obtaining M3 or better recovery. No patient grafted 12 months after injury recovered any useful function. Lesions of the posterior interosseous nerve had a consistently superior recovery. Power grip strength in the affected hand of patients averaged 60% of the unaffected hand while key pinch averaged 74%. There was good correlation between the Highet Scale rating of recovery and the ultimate power grip or key pinch strength obtained. Hand dexterity, as assessed by the turning and displacing tests of the Minnesota Rate of Manipulation Test, displayed a wide range of scores in both affected and unaffected hands. Nevertheless, a relative score derived from the results obtained in the displacing test did show correlation with the Highet Scale rating. All patients with M4 or better recovery obtained relative scores for the affected hand that were in the middle of the range of scores considered an average performance for a normal population. Patients who achieved M4 or better nerve recovery following radial nerve grafting also obtained a functional hand as evidenced by the results of grip, key pinch strength, and hand dexterity testing. Lesser degrees of recovery were accompanied by poorer strength and dexterity ratings reflecting inferior function. PMID- 9393663 TI - Warm ischaemia time in a model for small bowel transplantation. AB - Intestinal transplantation is associated with high rates of mortality and morbidity. This paper details our initial experience with 82 heterotopic small bowel transplants based upon the original rat model described by Monchik and Russell (Surgery 70:693-702, 1971). A key issue associated with mortality was a warm ischaemia time of more than 40 min (P < 0.01). Sixty-eight percent of the recipients (44/65) survived for more than 24 hr when the warm ischaemia time of the donor bowel was reduced to less than 40 min. Investigators establishing an animal model of heterotopic small bowel transplantation should pay particular attention to the warm ischaemia time of the donor bowel. PMID- 9393664 TI - Assessment of nerve ultrastructure by fibre-optic confocal microscopy. AB - Fibre-optic technology combined with confocality produces a microscope capable of optical thin sectioning. In this original study, tibial nerves have been stained in a rat model with a vital dye, 4-(4-diethylaminostyryl)-N-methylpyridinium iodide, and analysed by fibre-optic confocal microscopy to produce detailed images of nerve ultrastructure. Schwann cells, nodes of Ranvier and longitudinal myelinated sheaths enclosing axons were clearly visible. Single axons appeared as brightly staining longitudinal structures. This allowed easy tracing of multiple signal axons within the nerve tissue. An accurate measurement of internodal lengths was easily accomplished. This technique is comparable to current histological techniques, but does not require biopsy, thin sectioning or tissue fixing. This study offers a standard for further in vivo microscopy, including the possibility of monitoring the progression of nerve regeneration following microsurgical neurorraphy. PMID- 9393665 TI - Latissimus dorsi free flap for sacral wound closure: the world's longest vein grafts for free tissue transfer. AB - In 1983, Salibian et al. reported the use of a two-stage latissimus dorsi free tissue transfer to cover a sacral radiation ulcer using 28-centimeter thoracodorsal interposition vein grafts. In 1985, Nahai and Hagerty reported a similar case in which the procedure was performed in one stage with 25-centimeter vein grafts. We present a case in which a large sacral osteoradionecrosis ulcer is closed using this one-stage technique with 46-centimeter vein grafts, the longest ever reported for free tissue transfer. PMID- 9393666 TI - Ischemia increases the angiogenic potency of basic fibroblast growth factor (FGF 2). AB - The aim of this study was to investigate the angiogenic response to exogenously administered basic fibroblast growth factor (FGF-2) in normal and ischemic skin, using the hairless mouse ear microcirculatory model. The hairless mouse ear is a well-established model for in vivo studies of skin microcirculation. Using this model, angiogenesis- and angiogenesis-associated changes in the microcirculation can be directly and continuously viewed and quantified in a variety of different experimental settings. To create ischemia in the mouse ear, all but one of the three to four feeding vessels nourishing the ear were ligated 3 days prior to a local subdermal injection of FGF-2 (9.3 + 1-0.5 mm/mm2) or saline into the dorsum of the ears. Angiogenesis was quantified by direct observation, at high magnification, of the injection site where increases in total vessel length (TVL) were measured repeatedly over 18 days following injection. We found a significant (P < 0.01) increase in TVL in normal and ischemic ears injected with FGF-2. Saline injection also induced a significant increase in TVL in ischemic ears. However, the angiogenic response to FGF-2 in ischemic ears was significantly stronger than saline alone in ischemic ears or saline or FGF-2 in normal ears. This response could be used clinically to accelerate angiogenesis and thus increase perfusion in ischemic tissue. PMID- 9393667 TI - Vascularized bone grafts for congenital pseudarthrosis of the tibia. AB - Eight vascularized fibula grafts and two vascularized rib grafts were used for the treatment of 10 Boyd's Type II congenital pseudarthrosis of the tibia. All but one vascularized fibula graft united within 4 months. The two vascularized rib grafts did not unite until receiving a conventional bone graft. Nine spontaneous fractures were seen in four patients; all were subsequently treated successfully with cast or conventional bone graft. Corrective osteotomies were done in two patients. Follow-up averaged 8 years and 5 months (range, 5 years and 1 month to 14 years and 4 months). Average age at end of follow-up was 13 years and 6 months (range, 7 years and 10 months to 20 years and 4 months). After bony union was achieved, shortening of the affected leg averaged 3.8 centimeters, flexion deformity averaged 20 degrees, and valgus deformity averaged 24 degrees. In three patients, whose leg discrepancy averaged 4.9 centimeters, the leg was lengthened at an average patient age of 13 years and 9 months (age range, 11 years and 7 months to 15 years and 2 months). The resulting limb length discrepancy averaged 2.2 centimeters. Vascularized bone grafting is a reliable technique for achieving bony union in congenital pseudarthrosis of the tibia. Residual shortening may be corrected later by limb lengthening. PMID- 9393668 TI - Application of a new sleeve anastomosis technique to graft rearterialization in rat liver transplantation. AB - A new rearterialized orthotopic liver transplant (OLT) model in the rat is described. The model involved performing a novel sleeve anastomosis technique for graft rearterialization which consisted of three extraluminal suture anastomoses between the recipient's proper and the donor's common hepatic artery. The total surgical time and in particular the time required to perform the arterial anastomosis was significantly reduced with the utilization of this technique. Hepatic artery patency was 100% and the 4-week survival rate was greater than 90%. Applying this novel sleeve anastomosis technique not only simplifies the rearterialized rat OLT but also ensures that blood flow to all vascular beds is undisturbed. PMID- 9393669 TI - New strategies for chemokine inhibition and modulation: you take the high road and I'll take the low road. AB - Chemokines are low molecular weight cytokines that induce extravasation, chemotaxis, and activation of a wide variety of leukocytes. Members of the different chemokine families are defined by the orientation of specific critical cysteine residues, and are designated as C-X-C (e.g. interleukin-8), C-C (e.g. regulated upon activation normally T cell expressed and secreted, RANTES), or C (lymphotactin). All chemokines bind to members of a G-protein coupled serpentine receptor superfamily that span the leukocyte cell surface membrane seven times and mediate the biological activities of the individual ligands. Most chemokines possess two major binding surfaces: a high affinity site responsible for specific ligand/receptor interactions and a lower affinity site, also called the heparin binding or glycosaminoglycan-binding domain, believed to be responsible for the establishment and presentation of chemokine gradients on the surface of endothelial cells and within the extracellular matrix. Although chemokines are clearly beneficial in wound healing, hemopoiesis, and the clearance of infectious organisms, the continued expression of chemokines is associated with chronic inflammation. Therefore, this class of cytokines are attractive targets for the creation of antagonists that abrogate one or more chemokine functions. It is envisioned that such antagonists could serve as a new class of anti-inflammatory drugs. In this commentary, we will discuss two different but related strategies for antagonizing chemokine-induced functions, namely, disruption of the low and high affinity binding sites. PMID- 9393670 TI - Inhibition of inducible nitric oxide synthase gene expression and enzyme activity by epigallocatechin gallate, a natural product from green tea. AB - Chronic inflammation has been implicated as the underlying factor in the pathogenesis of many disorders. In the past decade, inflammation-related endogenous production of reactive nitrogen species, similar to oxygen free radicals, has also been suggested as a risk factor for cancer, in addition to the well-studied exogenous nitroso compounds. Epidemiological, in vitro, and animal model studies have implicated green tea to be protective against nitroso compound induced and inflammation-related cancer. Therefore, we investigated the effect of epigallocatechin-3-gallate (EGCG), one of the known biologically active catechins contained in green tea, on the production of nitric oxide (NO.). We have shown previously that EGCG reduces NO. production as measured by nitrite accumulation in the culture medium. Expanding on this finding, in this report we show that EGCG may do so by two mechanisms: reduction of inducible nitric oxide synthase (iNOS) gene expression and inhibition of enzyme activity. Addition of 1-10 microM EGCG to lipopolysaccharide- and interferon-gamma-activated mouse peritoneal cells reduced iNOS mRNA expression concentration dependently, to 82-14%, as measured by relative reverse transcription-polymerase chain reaction. Addition of 50-750 microM EGCG, in a concentration-dependent manner, inhibited the enzyme activity of iNOS, to 85-14%, and neuronal nitric oxide synthase (nNOS), to 93-56%, as measured by citrulline formation. EGCG competitively inhibited binding of arginine and tetrahydrobiopterin, and the gallate structure is important for this action. PMID- 9393671 TI - Sustained increase in intracellular free calcium and activation of cyclooxygenase 2 expression in mouse hepatoma cells treated with dioxin. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a non-genotoxic environmental pollutant that causes multiple adverse effects in experimental animals and in humans. We show here that TCDD treatment of mouse hepatoma cells causes a rapid mobilization of intracellular calcium both in wild type Hepa-1 cells and in its c2 variant, a cell line that has highly reduced levels of functional aromatic hydrocarbon (Ah) receptor (AHR). In wild type cells, but not in the c2 variant, TCDD treatment leads to a sustained elevation of cytosolic free calcium. TCDD also induces elevated levels of cyclooxygenase-2 (COX-2) mRNA in wild type and in c37, a CYP1A1-deficient cell line, but not in c2 cells. Induction of Cox-2 is in fact dependent on the presence of a functional Ah receptor, since it can be blocked by antisense oligonucleotides to Ah receptor mRNA. Most likely as a consequence of Cox-2 induction, we find a significant increase in the level of 12 hydroxyheptadecatrienoic acid (12-HHT) secreted from TCDD-treated Hepa-1 cells. In addition, we observe elevated levels of 6-keto prostaglandin F1alpha in c2 cells and high levels of secreted prostaglandin F2alpha in c2, c37 and c4, the variant cell line lacking aromatic hydrocarbon nuclear translocator protein. These data suggest that Cox-2 activation by TCDD leads to the release of prostaglandins, eicosanoids and other mediators which may have an important role in the biological and toxic effects of TCDD. PMID- 9393672 TI - Emergence of different mechanisms of resistance in the evolution of multidrug resistance in murine erythroleukemia cell lines. AB - We examined the genetic and biochemical bases for drug resistance and the order of appearance of different mechanisms underlying the increasingly more resistant murine erythroleukemia cell lines established in Adriamycin (ADR). In the first step low-level resistant cell line PC4-A5 (able to grow in 5 ng/mL ADR), there was a 2-fold reduction in topoisomerase IIalpha and topoisomerase IIbeta mRNA levels, as well as topoisomerase IIalpha protein and activity levels as compared with the parental cell line. The topoisomerase IIalpha activity levels remained reduced as the cells became increasingly more resistant. In contrast, the topoisomerase II mRNA and protein levels returned to approximately the parental levels in resistant cells growing in higher drug concentrations (40-160 ng/mL). Parental cells expressed the multidrug resistance protein (MRP), but beginning with PC4-A5 MRP expression decreased and remained reduced in increasingly resistant cell lines. At high levels of ADR resistance, the cells expressed the mdr3 gene concomitant with the appearance of vincristine resistance and energy dependent daunomycin and vincristine efflux. Glutathione levels, internal pH, and expression of the major vault protein (MVP) remained unchanged in all cell lines. Fluorescence microscopy revealed no alterations in daunomycin distribution or vesicle numbers between the parental and resistant cell lines. Different resistance mechanisms emerge sequentially as cells become more resistant to ADR; the mechanisms are retained during the development of multidrug resistance (MDR). In intermediate-level MDR cell lines (PC4-A10 and PC4-A20), resistance involves an as yet undetermined mechanism(s). PMID- 9393673 TI - Dinitrosyl-dithiol-iron complexes, nitric oxide (NO) carriers in vivo, as potent inhibitors of human glutathione reductase and glutathione-S-transferase. AB - Human glutathione reductase (GR) and rat liver glutathione-S-transferases (GSTs) had been shown to be inhibited by the nitric oxide (NO) carrier S-nitroso glutathione (GSNO). We have now extended these studies by measuring the effects of dinitrosyl-iron complexed thiols (DNIC-[RSH]2) on human GR, GST and glutathione peroxidase. DNIC-[RSH]2 represent important transport forms of NO but also of iron ions and glutathione in vivo. Human GR was found to be inhibited by dinitrosyl-iron-di-glutathione (DNIC-[GSH]2) and dinitrosyl-iron-di-L-cysteine (DNIC-Cys2) in two ways: both compounds were competitive with glutathione disulfide (GSSG), the inhibition constant (Ki) for reversible competition of DNIC [GSH]2 with GSSG being approximately 5 microM; preincubating GR for 10 min with 4 microM DNIC-[GSH]2 and 40 microM DNIC-Cys2, respectively, led to 50% irreversible enzyme inactivation. More than 95% GR inactivation was achieved by incubation with 36 microM DNIC-[GSH]2 for 30 min. This inhibition depended on the presence of NADPH. Absorption spectra of inhibited GR showed that the charge-transfer interaction between the isoalloxazine moiety of the prosthetic group flavin adenine dinucleotide (FAD) and the active site thiol Cys63 is disturbed by the modification. Cys2 and FAD could be ruled out as sites of the modification. Isolated human placenta glutathione-S-transferase and GST activity measured in hemolysates were also inhibited by DNIC-[GSH]2. This inhibition, however, was reversible and competitive with reduced glutathione, the Ki being 20 nM. The inhibition of GST induced by GSNO was competitive with reduced glutathione (GSH) (Ki = 180 microM) and with the second substrate of the reaction, 1-chloro-2,4, dinitrobenzene (Ki = 170 microM). An inhibition of human glutathione peroxidase by GSNO or DNIC-[RSH]2 was not detectable. Inactivation of GR by DNIC-[GSH]2 is by two orders of magnitude more effective than modification by GSNO; this result and the very efficient inhibition of GST point to a role of DNIC-[RSH]2 in glutathione metabolism. PMID- 9393674 TI - Synthesis and characterization of 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene (BODIPY)-labeled fluorescent ligands for the mu opioid receptor. AB - A series of opioid ligands utilizing the 4,4-difluoro-4-bora-3a,4a-diaza-s indacene (BODIPY) fluorophores 4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s indacene++ +-3-propionic acid or 4,4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-bora 3a,4a-diaza- s-indacene-3-propionic acid were synthesized and characterized for their ability to act as a suitable fluorescent label for the mu opioid receptor. All compounds displaced the mu opioid receptor binding of [3H]Tyr-D-Ala-Gly (Me)Phe-Gly-ol in monkey brain membranes with high affinity. The binding of fluorescent ligands to delta and kappa receptors was highly variable. 5,7 Dimethyl-BODIPY naltrexamine, "6-BNX," displayed subnanomolar affinities for the mu and kappa opioid receptors (Ki 0.07 and 0.43 nM, respectively) and nanomolar affinity at the delta (Ki 1.4 nM) receptor. Using fluorescence spectroscopy, the binding of 6-BNX in membranes from C6 glioma cells transfected with the cloned mu opioid receptor was investigated. In these membranes containing a high receptor density (10-80 pmol/mg protein), 6-BNX labeling was saturable, mu opioid specific, stereoselective (as determined with the isomers dextrorphan and levorphanol), and more than 90% specific. The results describe a series of newly developed fluorescent ligands for the mu opioid receptor and the use of one of these ligands as a label for the cloned mu receptor. These ligands provide a new approach for studying the structural and biophysical nature of opioid receptors. PMID- 9393675 TI - Pretranslational and posttranslational regulation of rat hepatic CYPs 3A2 and 2E1 by disulfiram. AB - The aldehyde dehydrogenase inhibitor disulfiram (DS) has been used to deter drinking in alcoholics, but it also precipitates pharmacokinetic interactions with coadministered drugs. From previous experiments conducted in vitro, it has been proposed that the ethanol-inducible cytochrome P450 2E1 (CYP2E1) is the major target for inhibition by DS, but the inference from reported drug interactions is that the drug inhibits multiple CYPs. The aim of the present study was to evaluate the inhibition of major constitutive CYPs in rat liver by DS. Thus, the effects of DS on activities mediated by CYPs 2A1/2, 2C11, 2E1, and 3A, which constitute approximately 80% of total CYPs in male rat liver, were evaluated. It was found that CYP2E1-mediated aniline 4-hydroxylase activity was weakly inhibited by DS in vitro, but that preincubation of the drug with NADPH supplemented microsomes to generate metabolites of DS enhanced the extent of inhibition somewhat. In contrast, constitutive testosterone hydroxylases were inhibited effectively at low concentrations of DS (20 microM decreased the activities of all hydroxylation pathways to 40-60% of control), and a preincubation step between DS and NADPH-fortified microsomes enhanced the inhibition of CYP2C11 and 3A2 activities. In vivo studies were undertaken in which a single dose of DS (100 mg/kg, i.p.) was administered to rats; 24 hr later, CYP2E1-mediated aniline 4-hydroxylase activity was decreased to about 50% of the activity in untreated control rats. CYP2E1 apoprotein and mRNA were also decreased to 38% of the respective control, and CYP3A apoprotein and CYP3A2 mRNA responded similarly. In contrast, CYP2C11 apoprotein was decreased to 66% of control after DS administration, and CYP2A1 expression was unchanged. These findings establish that multiple CYPs are targets for inhibition by DS and provide a basis for clinically significant drug interactions involving CYPs other than 2E1. In addition, the in vivo modulation of CYP function by DS administration is not restricted to enzyme inactivation and may also include down regulatory effects mediated at a pretranslational level. PMID- 9393676 TI - The hydrolysis of phosphatidyl-alcohols by phospholipases A2: effect of head group size and polarity. AB - The ability of a variety of secretory phospholipases A2 (sPLA2: EC 3.1.1.4) to bind to and hydrolyse a series of phosphatidyl-alcohol substrates, in the absence of detergent, was explored by both fluorescence-based kinetic and interfacial binding assays. The enzymes used were sPLA2 from porcine pancreas, Naja naja venom and a recombinant human non-pancreatic enzyme. Four dioleoyl phosphatidyl alcohols were used with different headgroups, methanol, ethanol, propanol and butanol. Comparative kinetic analyses with dioleoyl phosphatidyl-choline, dioleoyl phosphatidyl-glycerol and wheat germ phosphatidyl-inositol are also described. With the phosphatidyl-alcohol series, as the headgroup acyl-chain length increased the susceptibility to hydrolysis decreased. This effect was much more pronounced with the human non-pancreatic and the Naja naja venom enzymes than with the pancreatic enzyme. Maximum activity in this assay system was observed with porcine pancreatic sPLA2 and dioleoyl phosphatidyl-methanol (1440 +/- 167 micromol/min/mg). We demonstrate that the slow rate of hydrolysis of dioleoyl phosphatidyl-propanol by the human non-pancreatic secretory enzyme (4.56 +/- 0.90 micromol/min/mg) is not due to a lack of interfacial binding. The hydrolysis of mixtures of dioleoyl phosphatidyl-choline and dioleoyl phosphatidyl propanol in various molar proportions by Naja naja sPLA2 suggests good mixing of the two phospholipids with minimal phospholipid domain formation under these assay conditions. We present strong evidence for a stimulation of hydrolysis of phosphatidyl-choline by human non-pancreatic sPLA2 in the presence of as little as 1 mol% phosphatidyl-methanol (<40 fold total rate enhancement). Overall, the results demonstrate that the rates of hydrolysis of anionic phospholipids by sPLA2 vary considerably with the different enzymes from this close structurally related family. The tight binding of the human enzyme to poorly hydrolysable anionic phospholipid vesicles provides a novel mechanism of enzyme inhibition by interfacial sequestration. PMID- 9393677 TI - Renal cellular transport, metabolism, and cytotoxicity of S-(6 purinyl)glutathione, a prodrug of 6-mercaptopurine, and analogues. AB - The disposition of S-(6-purinyl)glutathione (6-PG) and its metabolites, including the antitumor agent 6-mercaptopurine (6-MP), was characterized in freshly isolated renal cortical cells from male F344 rats to assess the ability of the kidney to convert 6-PG to 6-MP. The intracellular transport and accumulation of 6 PG and 6-MP, the metabolism of 6-PG to 6-MP, and the potential cytotoxicity of 6 MP, 6-thioxanthine (6-ThXan), and 6-thioguanine (6-ThGua) were determined. 6-PG and 6-MP were accumulated by renal cortical cells by time- and concentration dependent processes, reaching maximal levels of 14.2 and 1.52 nmol/10(6) cells, respectively, with 1 mM concentrations of each compound. Treatment with acivicin, an inhibitor of 6-PG metabolism by gamma-glutamyltransferase, increased accumulation of 6-PG, and treatment with alpha-keto-gamma-methiolbutyrate, a keto acid cosubstrate that stimulates activity of the cysteine conjugate beta-lyase (beta-lyase), which generates 6-MP, decreased accumulation of 6-PG. Incubation of renal cells with 10 mM 6-PG generated 6-MP at a rate of 2.4 nmol/min per 10(6) cells, demonstrating that the beta-lyase pathway forms the desired product from the prodrug within the intact renal cell. Preincubation of cells with acivicin or aminooxyacetic acid, an inhibitor of the beta-lyase, decreased the net formation of 6-MP, demonstrating further the function of the beta-lyase. 6-MP, 6-ThXan, and 6-ThGua exhibited approximately equivalent cytotoxicity (45-55% release of lactate dehydrogenase with 1 mM at 2 hr) in isolated renal cells. Based on the known antitumor potency of these agents, this suggests that cytotoxicity and antitumor activity occur by distinct mechanisms. The high amount of accumulation of 6-PG and its subsequent metabolism to 6-MP, as compared with the relatively low amount of accumulation of 6-MP, in renal cells suggest that 6-PG can function as a prodrug and is a more effective delivery vehicle for 6-MP to renal cells than 6-MP itself. Administration of 6-PG may be an effective means of treating renal tumors or suppressing renal transplant rejection. PMID- 9393678 TI - Receptor-stimulated phospholipase C activity in human umbilical artery cultured endothelial cells grown in a low oxygen environment. AB - Endothelial cells of the human umbilical blood vessels are widely cultured in an oxygen tension (21%) far above that in which they exist in vivo (3%). This study investigates the effect of the long term culture (ca. 1 month) of human umbilical artery endothelial cells in a reduced oxygen environment (3%: HUAEC3) in comparison to cells grown in a 'normoxic' environment (21%: HUAEC21). Despite reports of altered metabolic pathways and reduced membrane integrity in other cell types, the characteristics of HUAEC3 were found to be similar to those of HUAEC21 with respect to morphology, immunocytochemical profile and in vitro growth rates. Cellular glutathione was maintained in these cells although ATP levels in HUAEC3 were found to be significantly lower than those observed in HUAEC21. The phosphoinositide responses of the HUAEC3 to a variety of agonists were also found to be of similar magnitude to those observed in HUAEC21. In addition, the pharmacological characteristics of the phospholipase C-linked histamine H1 and P2y2 (P2U) receptors were not changed by culture of cells in a low oxygen environment. PMID- 9393679 TI - Monoamine oxidase inhibitory properties of some methoxylated and alkylthio amphetamine derivatives: structure-activity relationships. AB - The monoamine oxidase (MAO) inhibitory properties of a series of amphetamine derivatives with different substituents at or around the para position of the aromatic ring were evaluated. In in vitro studies in which a crude rat brain mitochondrial suspension was used as the source of MAO, several compounds showed a strong (IC50 in the submicromolar range), selective, reversible, time independent, and concentration-related inhibition of MAO-A. After i.p. injection, the compounds induced an increase of serotonin and a decrease of 5 hydroxyindoleacetic acid in the raphe nuclei and hippocampus, confirming the in vitro results. The analysis of structure-activity relationships indicates that: molecules with amphetamine-like structure and different substitutions on the aromatic ring are potentially MAO-A inhibitors; substituents at different positions of the aromatic ring modify the potency but have little influence on the selectivity; substituents at the para position such as amino, alkoxyl, halogens, or alkylthio produce a significant increase in the activity; the para substituent must be an electron donor; bulky groups next to the para substituent lead to a decrease in the activity; substituents located at positions more distant on the aromatic ring have less influence and, even when the substituent is a halogen (Cl, Br), an increase in the activity of the compound is obtained. Finally, the MAO-A inhibitory properties of some of the compounds evaluated are discussed in relation to: (a) potential antidepressant activity, and (b) their reported hallucinogenic, neurotoxic, or anxiolytic effects. PMID- 9393680 TI - High sensitivity of leukemic peripheral blood lymphocytes to triethyllead action. AB - In a previous study we reported that triethyllead (Et3Pb+) inhibits cell proliferation of normal human lymphocytes. To further characterize this interaction, we studied herein the effects of Et3Pb+ on the cell viability of normal and leukemic human lymphocytes and analysed the expression and dynamics of the monomer/polymer equilibrium of tubulin in these cells. Short- and long-term cell culture experiments demonstrated significantly different dose-dependent effects of Et3Pb+ on cell viability of leukemic compared to normal lymphocytes. Indeed, in the presence of increasing concentrations of Et3Pb+ (10(-12)-10(-5) M), primary cultures of chronic lymphocytes (CLL) and acute lymphoblastic (ALL) leukemic human Lymphocytes were much more sensitive to Et3Pb+ treatment when compared to normal peripheral blood lymphocytes (PBL). The IC50 values were approximately 5 x 10(-6) M for PBL and 8 x 10(-10) M for both CLL and ALL respectively, when cells were preincubated for 3 h with this agent. These experiments revealed a 1000-fold higher responsiveness of leukemic cells to Et3Pb+ treatment. Quantitative immunoblot analysis showed that leukemic cells express up to 4-fold higher total tubulin amounts. However, the proportion of polymerized tubulin in leukemic compared to normal lymphocytes increased only slightly (up to 1.4-fold). These findings reveal a significant decrease in the polymeric to total tubulin ratio in leukemic lymphocytes, indicating important modifications in tubulin dynamics and reorganization of the microtubular structures. Our results demonstrate that leukemic cells are much more sensitive than normal lymphocytes to Et3Pb+ action. This effect may be due to the altered monomer/polymer dynamic equilibrium of tubulin shown in leukemic cells. It is, therefore, worthwhile exploring future applied uses of Et3Pb+ as a potential suppressor of leukemic cell growth. PMID- 9393681 TI - Transcriptional and translational control of the genes for the mating pair formation apparatus of promiscuous IncP plasmids. AB - The trb operon of broad-host-range plasmid RK2 encodes most of the genes required for formation of mating-pair apparatus and is thus essential for the promiscuous spread of this plasmid. Only two promoters, lying upstream of trbA and trbB, have been identified for this operon. trbB encodes a protein belonging to a large family of proteins which function in the assembly of apparatuses associated with the cell surface. trbA encodes a repressor protein, one of whose targets is the trbB promoter. trbAp is arranged as a face-to-face divergent promoter with trfAp, the strongest of the three promoters in this region. trfAp completely inhibits trbAp unless it is repressed by the KorA protein, a key regulator encoded in the plasmid's central control operon. We show that when trfAp is firing constitutively, it also appears to interfere with trbBp, but that trbBp activity increases when trfAp activity is decreased by repression or mutation. A second global regulator encoded in the central control operon, KorB, represses trbBp, trfAp, and trbAp. The results presented here show that both KorB and TrbA are necessary for full repression of trbBp. The region between trbA and trbB encodes a large inverted repeat which has been proposed to modulate translation of trbB on transcripts which are initiated at trbAp but not trbBp. Using translational fusions to lacZ, we show that translation of trbB is completely blocked when transcripts incorporate the inverted repeat upstream of trbB but proceeds with reasonable efficiency when deletions remove the sequences predicted to sequester the ribosome binding site. Results from both transcriptional fusion and direct measurement of transcript size and intensity by Northern blot analysis show that most trbA transcripts are monocistronic and serve to express only trbA, although some transcription continues into trbB. The monocistronic trbA transcript appears to be the result of transcription termination downstream of trbA. Thus, trbAp and trbA appear to form an operon distinct from the trbB-trbP operon. Consequently, trbA and the switch that controls its expression help to provide the sequential steps which allow efficient expression of transfer genes during plasmid establishment but tight repression once the plasmid is established. PMID- 9393682 TI - The Candida albicans CDR3 gene codes for an opaque-phase ABC transporter. AB - We report the cloning and functional analysis of a third member of the CDR gene family in Candida albicans, named CDR3. This gene codes for an ABC (ATP-binding cassette) transporter of 1,501 amino acids highly homologous to Cdr1p and Cdr2p (56 and 55% amino acid sequence identity, respectively), two transporters involved in fluconazole resistance in C. albicans. The predicted structure of Cdr3p is typical of the PDR/CDR family, with two similar halves, each comprising an N-terminal hydrophilic domain with consensus sequences for ATP binding and a C terminal hydrophobic domain with six predicted transmembrane segments. Northern analysis showed that CDR3 expression is regulated in a cell-type-specific manner, with low levels of CDR3 mRNA in CAI4 yeast and hyphal cells, high levels in WO-1 opaque cells, and undetectable levels in WO-1 white cells. Disruption of both alleles of CDR3 in CAI4 resulted in no obvious changes in cell morphology, growth rate, or susceptibility to fluconazole. Overexpression of Cdr3p in C. albicans did not result in increased cellular resistance to fluconazole, cycloheximide, and 4-nitroquinoline-N-oxide, which are known substrates for different transporters of the PDR/CDR family. These results indicate that despite a high degree of sequence conservation with C. albicans Cdr1p and Cdr2p, Cdr3p does not appear to be involved in drug resistance, at least to the compounds tested which include the clinically relevant antifungal agent fluconazole. Rather, the high level of Cdr3p expression in WO-1 opaque cells suggests an opaque-phase associated biological function which remains to be identified. PMID- 9393683 TI - Synergistic roles of HslVU and other ATP-dependent proteases in controlling in vivo turnover of sigma32 and abnormal proteins in Escherichia coli. AB - Production of abnormal proteins during steady-state growth induces the heat shock response by stabilizing normally unstable sigma32 (encoded by the rpoH gene) specifically required for transcription of heat shock genes. We report here that a multicopy plasmid carrying the hslVU operon encoding a novel ATP-dependent protease inhibits the heat shock response induced by production of human prourokinase (proUK) in Escherichia coli. The overproduction of HslVU (ClpQY) protease markedly reduced the stability and accumulation of proUK and thus reduced the induction of heat shock proteins. In agreement with this finding, deletion of the chromosomal hslVU genes significantly enhanced levels of proUK and sigma32 without appreciably affecting cell growth. When the deltahslVU deletion was combined with another protease mutation (lon, clpP, or ftsH/hflB), the resulting multiple mutations caused higher stabilization of proUK and sigma32, enhanced synthesis of heat shock proteins, and temperature-sensitive growth. Furthermore, overproduction of HslVU protease reduced sigma32 levels in strains that were otherwise expected to produce enhanced levels of sigma32 due either to the absence of Lon-ClpXP proteases or to the limiting levels of FtsH protease. Thus, a set of ATP-dependent proteases appear to play synergistic roles in the negative control of the heat shock response by modulating in vivo turnover of sigma32 as well as through degradation of abnormal proteins. PMID- 9393684 TI - Characterization of the Rhizobium (Sinorhizobium) meliloti high- and low-affinity phosphate uptake systems. AB - Genetic studies have suggested that Rhizobium (Sinorhizobium) meliloti contains two distinct phosphate (Pi) transport systems, encoded by the phoCDET genes and the orfA-pit genes, respectively. Here we present data which show that the ABC type PhoCDET system has a high affinity for Pi (Km, 0.2 microM) and that Pi uptake by this system is severely inhibited by phosphonates. This high-affinity uptake system was induced under Pi-limiting conditions and was repressed in the presence of excess Pi. Uptake via the OrfA-Pit system was examined in (i) a phoC mutant which showed increased expression of the orfA-pit genes as a result of a promoter-up mutation and (ii) a phoB mutant (PhoB is required for phoCDET expression). Pi uptake in both strains exhibited saturation kinetics (Km, 1 to 2 microM) and was not inhibited by phosphonates. This uptake system was active in wild-type cells grown with excess Pi and appeared to be repressed when the cells were starved for Pi. Thus, our biochemical data show that the OrfA-Pit and PhoCDET uptake systems are differentially expressed depending on the state of the cell with respect to phosphate availability. PMID- 9393686 TI - Glycolytic flux is conditionally correlated with ATP concentration in Saccharomyces cerevisiae: a chemostat study under carbon- or nitrogen-limiting conditions. AB - Anaerobic and aerobic chemostat cultures of Saccharomyces cerevisiae were performed at a constant dilution rate of 0.10 h(-1). The glucose concentration was kept constant, whereas the nitrogen concentration was gradually decreasing; i.e., the conditions were changed from glucose and energy limitation to nitrogen limitation and energy excess. This experimental setup enabled the glycolytic rate to be separated from the growth rate. There was an extensive uncoupling between anabolic energy requirements and catabolic energy production when the energy source was present in excess both aerobically and anaerobically. To increase the catabolic activity even further, experiments were carried out in the presence of 5 mM acetic acid or benzoic acid. However, there was almost no effect with acetate addition, whereas both respiratory (aerobically) and fermentative activities were elevated in the presence of benzoic acid. There was a strong negative correlation between glycolytic flux and intracellular ATP content; i.e., the higher the ATP content, the lower the rate of glycolysis. No correlation could be found with the other nucleotides tested (ADP, GTP, and UTP) or with the ATP/ADP ratio. Furthermore, a higher rate of glycolysis was not accompanied by an increasing level of glycolytic enzymes. On the contrary, the glycolytic enzymes decreased with increasing flux. The most pronounced reduction was obtained for HXK2 and ENO1. There was also a correlation between the extent of carbohydrate accumulation and glycolytic flux. A high accumulation was obtained at low glycolytic rates under glucose limitation, whereas nitrogen limitation during conditions of excess carbon and energy resulted in more or less complete depletion of intracellular storage carbohydrates irrespective of anaerobic or aerobic conditions. However, there was one difference in that glycogen dominated anaerobically whereas under aerobic conditions, trehalose was the major carbohydrate accumulated. Possible mechanisms which may explain the strong correlation between glycolytic flux, storage carbohydrate accumulation, and ATP concentrations are discussed. PMID- 9393685 TI - Host cell phospholipids are trafficked to and then modified by Chlamydia trachomatis. AB - There is little information on the trafficking of eukaryotic lipids from a host cell to either the cytoplasmic membrane of or the vacuolar membrane surrounding intracellular pathogens. Purified Chlamydia trachomatis, an obligate intracellular bacterial parasite, contains several eukaryotic glycerophospholipids, yet attempts to demonstrate transfer of these lipids to the chlamydial cell membrane have not been successful. In this report, we demonstrate that eukaryotic glycerophospholipids are trafficked from the host cell to C. trachomatis. Phospholipid trafficking was assessed by monitoring the incorporation of radiolabelled isoleucine, a precursor of C. trachomatis specific branched-chain fatty acids, into host-derived glycerophospholipids and by monitoring the transfer of host phosphatidylserine to chlamydiae and its subsequent decarboxylation to form phosphatidylethanolamine. Phospholipid trafficking to chlamydiae was unaffected by brefeldin A, an inhibitor of Golgi function. Furthermore, no changes in trafficking were observed when C. trachomatis was grown in a mutant cell line with a nonfunctional, nonspecific phospholipid transfer protein. Host glycerophospholipids are modified by C. trachomatis, such that a host-synthesized straight-chain fatty acid is replaced with a chlamydia-synthesized branched-chain fatty acid. We also demonstrate that despite the acquisition of host-derived phospholipids, C. trachomatis is capable of de novo synthesis of phospholipids typically synthesized by prokaryotic cells. Our results provide novel information on chlamydial phospholipid metabolism and eukaryotic cell lipid trafficking, and they increase our understanding of the evolutionary steps leading to the establishment of an intimate metabolic association between an obligate intracellular bacterial parasite and a eukaryotic host cell. PMID- 9393687 TI - Expression of a stress- and starvation-induced dps/pexB-homologous gene is controlled by the alternative sigma factor sigmaB in Bacillus subtilis. AB - SigmaB-dependent general stress proteins (Gsps) of Bacillus subtilis are essential for the development of glucose-starvation-induced cross-resistance to oxidative challenge. However, the proteins directly involved in this nonspecific resistance to oxidative stress have to be identified. We found that one prominent Gsp displayed strong sequence similarity to the previously characterized oxidative-stress-inducible MrgA protein of B. subtilis and to the starvation induced Dps/PexB protein of Escherichia coli. We therefore designated this prominent Gsp Dps. While MrgA belongs to the peroxide-stress-inducible proteins needed for the H2O2-inducible adaptive response to oxidative stress, Dps belongs to the proteins induced by heat, salt, or ethanol stress and after starvation for glucose but not by a sublethal oxidative challenge. Primer extension experiments identified two overlapping promoters upstream of the coding region of dps, one being sigmaB dependent (PB) and the other being sigmaB independent (P1). Both promoters contribute to the basal level of dps during growth. After stress or during entry into the stationary phase, transcription from PB strongly increased whereas transcription from P1 decreased. Mutant strains lacking Dps completely failed to develop glucose-starvation-induced resistance to oxidative stress. These results confirm our suggestion that sigmaB-dependent general stress proteins of B. subtilis are absolutely required for the development of nonspecific resistance to oxidative stress. PMID- 9393688 TI - A gene (plsD) from Clostridium butyricum that functionally substitutes for the sn glycerol-3-phosphate acyltransferase gene (plsB) of Escherichia coli. AB - The sn-glycerol-3-phosphate acyltransferase (plsB) of Escherichia coli is a key regulatory enzyme that catalyzes the first committed step in phospholipid biosynthesis. We report the initial characterization of a novel gene (termed plsD) from Clostridium butyricum, cloned based on its ability to complement the sn-glycerol-3-phosphate auxotrophic phenotype of a plsB mutant strain of E. coli. Unlike the 83-kDa PlsB acyltransferase from E. coli, the predicted plsD open reading frame encoded a protein of 26.5 kDa. Two regions of strong homology to other lipid acyltransferases, including PlsB and PlsC analogs from mammals, plants, yeast, and bacteria, were identified. PlsD was most closely related to the 1-acyl-sn-glycerol-3-phosphate acyltransferase (plsC) gene family but did not complement the growth of plsC(Ts) mutants. An in vivo metabolic labeling experiment using a plsB plsX plsC(Ts) strain of E. coli confirmed that the plsD expression restored the ability of the cells to synthesize 1-acyl-glycerol-3 phosphate. However, glycerol-3-phosphate acyltransferase activity was not detected in vitro in assays using either acyl-acyl carrier protein or acyl coenzyme A as the substrate. PMID- 9393689 TI - Analysis of the fnrL gene and its function in Rhodobacter capsulatus. AB - The fnr gene encodes a regulatory protein involved in the response to oxygen in a variety of bacterial genera. For example, it was previously shown that the anoxygenic, photosynthetic bacterium Rhodobacter sphaeroides requires the fnrL gene for growth under anaerobic, photosynthetic conditions. Additionally, the FnrL protein in R. sphaeroides is required for anaerobic growth in the dark with an alternative electron acceptor, but it is not essential for aerobic growth. In this study, the fnrL locus from Rhodobacter capsulatus was cloned and sequenced. Surprisingly, an R. capsulatus strain with the fnrL gene deleted grows like the wild type under either photosynthetic or aerobic conditions but does not grow anaerobically with alternative electron acceptors such as dimethyl sulfoxide (DMSO) or trimethylamine oxide. It is demonstrated that the c-type cytochrome induced upon anaerobic growth on DMSO is not synthesized in the R. capsulatus fnrL mutant. In contrast to wild-type strains, R. sphaeroides and R. capsulatus fnrL mutants do not synthesize the anaerobically, DMSO-induced reductase. Mechanisms that explain the basis for FnrL function in both organisms are discussed. PMID- 9393690 TI - The TolB protein interacts with the porins of Escherichia coli. AB - TolB is a periplasmic protein of the cell envelope Tol complex. It is partially membrane associated through an interaction with the outer membrane lipoprotein PAL (peptidoglycan-associated lipoprotein), which also belongs to the Tol system. The interaction of TolB with outer membrane porins of Escherichia coli was investigated with a purified TolB derivative harboring a six-histidine tag. TolB interacted with the trimeric porins OmpF, OmpC, PhoE, and LamB but not with their denatured monomeric forms or OmpA. These interactions took place both in the presence and in the absence of lipopolysaccharide. TolA, an inner membrane component of the Tol system, also interacts with the trimeric porins via its central periplasmic domain (R. Derouiche, M. Gavioli, H. Benedetti, A. Prilipov, C. Lazdunski, and R. Lloubes, EMBO J. 15:6408-6415, 1996). In the presence of the purified central domain of TolA (TolAIIHis), the TolB-porin complexes disappeared to form TolAIIHis-porin complexes. These results suggest that the interactions of TolA and TolB with porins might take place in vivo and might be concomitant events participating in porin assembly. They also suggest that the Tol system as a whole may be involved in porin assembly in the outer membrane. PMID- 9393691 TI - Cloning and characterization of the aru genes encoding enzymes of the catabolic arginine succinyltransferase pathway in Pseudomonas aeruginosa. AB - The arginine succinyltransferase (AST) pathway is the major arginine and ornithine utilization (aru) pathway under aerobic conditions in Pseudomonas aeruginosa. A 26-kb DNA fragment of the P. aeruginosa PAO1 chromosome carrying the regulatory argR gene and the aru structural gene cluster was cloned. Complementation tests and nucleotide sequence data established the locations of the argR, aruC, aruF, aruG, aruD, aruB, and aruE genes, in that order. The aruR, aruC, aruD, aruB, and aruE genes specify the ArgR regulatory protein, N2 succinylornithine 5-aminotransferase, N-succinylglutamate 5-semialdehyde dehydrogenase, N2-succinylarginine dihydrolase, and N-succinylglutamate desuccinylase, respectively, and the aruF and aruG genes encode the subunits (AruAI and AruAII) of arginine and ornithine N2-succinyltransferases. Furthermore, in vivo analysis of transcriptional aru fusions and of polar insertion mutations located at different sites in the aru cluster indicated the presence of three transcriptional units which are controlled by ArgR. The aruCFGDB genes appear to form an operon transcribed from a promoter upstream of aruC, whereas aruE has its own promoter. The argR gene, which is located upstream of the aruCFGDB operon, is a member of another (aot) operon coding for arginine transport genes. The deduced amino acid sequences of the AST enzymes were compared to those of homologous proteins of Escherichia coli specified by the ast genes lying in the chromosome region from 39.2 to 39.5 min (Kohara clone 327; GenBank/EMBL/DDJB accession no. D90818). The overall organization of the aru and ast genes in both organisms is similar, with the exception that E. coli appears to have a single AST gene. PMID- 9393692 TI - Mutational analysis of the R64 oriT region: requirement for precise location of the NikA-binding sequence. AB - Conjugative DNA transfer of IncI1 plasmid R64 is initiated by the introduction of a site- and strand-specific nick into the origin of transfer (oriT). In R64 oriT, 17-bp (repeat A and B) and 8-bp inverted-repeat sequences with mismatches are located 8 bp away from the nick site. The nicking is mediated by R64 NikA and NikB proteins. To analyze the functional organization of the R64 oriT region, various deletion, insertion, and substitution mutations were introduced into a 92 bp minimal R64 oriT sequence and their effects on oriT function were investigated. This detailed analysis confirms our previous prediction that the R64 oriT region consists of an oriT core sequence and additional sequences necessary for full oriT activity. The oriT core sequence consists of the repeat A sequence, which is recognized by R64 NikA protein, and the nick region sequence, which is conserved among various origins of transfer and is most probably recognized by NikB protein. The oriT core sequence is sufficient for NikAB mediated oriT-specific nicking. Furthermore, it was shown that the repeat A sequence is essential for localization to a precise position relative to the nick site for oriT function. This seems to be required for the formation of a functional ternary complex consisting of NikA and NikB proteins and oriT DNA. The repeat B sequence and 8-bp inverted repeat sequences are suggested to be required for the termination of DNA transfer. PMID- 9393693 TI - Transcriptional regulation of type 4 pilin genes and the site-specific recombinase gene, piv, in Moraxella lacunata and Moraxella bovis. AB - Moraxella lacunata and Moraxella bovis use type 4 pili to adhere to epithelial tissues of the cornea and conjunctiva. Primer extension analyses were used to map the transcriptional start sites for the genes encoding the major pilin subunits (tfpQ/I) and the DNA invertase (piv), which determines pilin type expression. tfpQ/I transcription starts at a sigma54-dependent promoter (tfpQ/Ip2) and, under certain growth conditions, this transcription is accompanied by weaker upstream transcription that starts at a potential sigma70-dependent promoter (tfpQ/Ip1). piv is expressed in both M. lacunata and M. bovis from a putative sigma70 dependent promoter (pivp) under all conditions assayed. Sigma54-dependent promoters require activators in order to initiate transcription; therefore, it is likely that tfpQ/Ip2 is also regulated by an activator in Moraxella. Primer extension assays with RNA isolated from Escherichia coli containing the subcloned pilin inversion region from M. lacunata showed that pivp is used for the expression of piv; however, tfpQ/Ip2 is not used for the transcription of tfpQ/I. Transcription from tfpQ/Ip2 was activated in E. coli when the sensor (PilS) and response regulator (PilR) proteins of type 4 pilin transcription in Pseudomonas aeruginosa were expressed from a plasmid. These results suggest that the expression of the type 4 pilin in M. lacunata and M. bovis is regulated not only by a site-specific DNA inversion system but also by a regulatory system which is functionally analogous to the PilS-PilR two-component system of P. aeruginosa. PMID- 9393694 TI - Cloning, sequencing, and expression of the gene encoding Clostridium paraputrificum chitinase ChiB and analysis of the functions of novel cadherin like domains and a chitin-binding domain. AB - The Clostridium paraputrificum chiB gene, encoding chitinase B (ChiB), consists of an open reading frame of 2,493 nucleotides and encodes 831 amino acids with a deduced molecular weight of 90,020. The deduced ChiB is a modular enzyme composed of a family 18 catalytic domain responsible for chitinase activity, two reiterated domains of unknown function, and a chitin-binding domain (CBD). The reiterated domains are similar to the repeating units of cadherin proteins but not to fibronectin type III domains, and therefore they are referred to as cadherin-like domains. ChiB was purified from the periplasm fraction of Escherichia coli harboring the chiB gene. The molecular weight of the purified ChiB (87,000) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS PAGE) analysis, was in good agreement with the value (86,578) calculated from the deduced amino acid sequence excluding the signal peptide. ChiB was active toward chitin from crab shells, colloidal chitin, glycol chitin, and 4 methylumbelliferyl beta-D-N,N'-diacetylchitobioside [4-MU-(GlcNAc)2]. The pH and temperature optima of the enzyme were 6.0 and 45 degrees C, respectively. The Km and Vmax values for 4-MU-(GlcNAc)2 were estimated to be 6.3 microM and 46 micromol/min/mg, respectively. SDS-PAGE, zymogram, and Western blot analyses using antiserum raised against purified ChiB suggested that ChiB was one of the major chitinase species in the culture supernatant of C. paraputrificum. Deletion analysis showed clearly that the CBD of ChiB plays an important role in hydrolysis of native chitin but not processed chitin such as colloidal chitin. PMID- 9393695 TI - In vitro Tn7 mutagenesis of Haemophilus influenzae Rd and characterization of the role of atpA in transformation. AB - Haemophilus influenzae Rd is a gram-negative bacterium capable of natural DNA transformation. The competent state occurs naturally in late exponential growth or can be induced by a nutritional downshift or by transient anaerobiosis. The genes cya, crp, topA, and sxy (tfoX) are known to function in the regulation of competence development. The phosphoenolpyruvate:carbohydrate phosphotransferase system functions to maintain levels of cyclic AMP necessary for competence development but is not directly involved in regulation. The exact signal(s) for competence and the genes that mediate the signal(s) are still unknown. In an effort to find additional regulatory genes, H. influenzae Rd was mutated by using an in vitro Tn7 system and screened for mutants with a reduced ability to induce the competence-regulatory gene, comA. Insertions in atpA, a gene coding for the alpha subunit of the F1 cytoplasmic domain of the ATP synthase, reduce transformation frequencies about 20-fold and cause a significant reduction in expression of competence-regulatory genes, while the expression of constitutive competence genes is only minimally affected. In addition, we found that an insertion in atpB, which encodes the a subunit of the F0 membrane-spanning domain, has a similar effect on transformation frequencies. PMID- 9393696 TI - Pectate lyase PelI of Erwinia chrysanthemi 3937 belongs to a new family. AB - Erwinia chrysanthemi 3937 secretes five major isoenzymes of pectate lyases encoded by the pel4, pelB, pelC, pelD, and pelE genes and a set of secondary pectate lyases, two of which, pelL and pelZ, have been already identified. We cloned the pelI gene, encoding a ninth pectate lyase of E. chrysanthemi 3937. The pelI reading frame is 1,035 bases long, corresponding to a protein of 344 amino acids including a typical amino-terminal signal sequence of 19 amino acids. The purified mature PelI protein has an isoelectric point of about 9 and an apparent molecular mass of 34 kDa. PelI has a preference for partially methyl esterified pectin and presents an endo-cleaving activity with an alkaline pH optimum and an absolute requirement for Ca2+ ions. PelI is an extracellular protein secreted by the Out secretory pathway of E. chrysanthemi. The PelI protein is very active in the maceration of plant tissues. A pelI mutant displayed reduced pathogenicity on chicory leaves, but its virulence did not appear to be affected on potato tubers or Saintpaulia ionantha plants. The pelI gene constitutes an independent transcriptional unit. As shown for the other pel genes, the transcription of pelI is dependent on various environmental conditions. It is induced by pectic catabolic products and affected by growth phase, oxygen limitation, temperature, nitrogen starvation, and catabolite repression. Regulation of pelI expression appeared to be dependent on the three repressors of pectinase synthesis, KdgR, PecS, and PecT, and on the global activator of sugar catabolism, cyclic AMP receptor protein. A functional KdgR binding site was identified close to the putative pelI promoter. Analysis of the amino acid sequence of PelI revealed high homology with a pectate lyase from Erwinia carotovora subsp. carotovora (65% identity) and low homology with pectate lyases of the phytopathogenic fungus Nectria haematococca (Fusarium solani). This finding indicates that PelI belongs to pectate lyase class III. Using immunoblotting experiments, we detected PelI homologs in various strains of E. chrysanthemi and E. carotovora subsp. carotovora but not in E. carotovora subsp. atroseptica. PMID- 9393697 TI - The cold shock response of the psychrotrophic bacterium Pseudomonas fragi involves four low-molecular-mass nucleic acid-binding proteins. AB - The psychrotrophic bacterium Pseudomonas fragi was subjected to cold shocks from 30 or 20 to 5 degrees C. The downshifts were followed by a lag phase before growth resumed at a characteristic 5 degrees C growth rate. The analysis of protein patterns by two-dimentional gel electrophoresis revealed overexpression of 25 or 17 proteins and underexpression of 12 proteins following the 30- or 20 to-5 degrees C shift, respectively. The two downshifts shared similar variations of synthesis of 20 proteins. The kinetic analysis distinguished the induced proteins into cold shock proteins (Csps), which were rapidly but transiently overexpressed, and cold acclimation proteins (Caps), which were more or less rapidly induced but still overexpressed several hours after the downshifts. Among the cold-induced proteins, four low-molecular-mass proteins, two of them previously characterized as Caps (CapA and CapB), and heat acclimation proteins (Haps) as well as heat shock proteins (Hsps) for the two others (TapA and TapB) displayed higher levels of induction. Partial amino acid sequences, obtained by microsequencing, were used to design primers to amplify by PCR the four genes and then determine their nucleotide sequences. A BamHI-EcoRI restriction fragment of 1.9 kb, containing the complete coding sequence for capB, was cloned and sequenced. The four peptides belong to the family of small nucleic acid-binding proteins as CspA, the major Escherichia coli Csp. They are likely to play a major role in the adaptative response of P. fragi to environmental temperature changes. PMID- 9393698 TI - A cysG mutant strain of Rhizobium etli pleiotropically defective in sulfate and nitrate assimilation. AB - By its inability to grow on sulfate as the sole sulfur source, a mutant strain (CTNUX8) of Rhizobium etli carrying Tn5 was isolated and characterized. Sequence analysis showed that Tn5 is inserted into a cysG (siroheme synthetase)-homologous gene. By RNase protection assays, it was established that the cysG-like gene had a basal level of expression in thiosulfate- or cysteine-grown cells, which was induced when sulfate or methionine was used. Unlike its wild-type parent (strain CE3), the mutant strain, CTNUX8, was also unable to grow on nitrate as the sole nitrogen source and was unable to induce a high level of nitrite reductase. Despite its pleiotropic phenotype, strain CTNUX8 was able to induce pink, effective (N2-fixing) nodules on the roots of Phaseolus vulgaris plants. However, mixed inoculation experiments showed that strain CTNUX8 is significantly different from the wild type in its ability to nodulate. Our data support the notion that sulfate (or sulfite) is the sulfur source of R. etli in the rhizosphere, while cysteine, methionine, or glutathione is supplied by the root cells to bacteria growing inside the plant. PMID- 9393699 TI - A null mutation in the Bacillus subtilis aconitase gene causes a block in Spo0A phosphate-dependent gene expression. AB - The citB gene of Bacillus subtilis encodes aconitase, the enzyme of the Krebs citric acid cycle, which is responsible for the interconversion of citrate and isocitrate. A B. subtilis strain with an insertion mutation in the citB gene was devoid of aconitase activity and aconitase protein, required glutamate for growth in minimal medium, and was unable to sporulate efficiently in nutrient broth sporulation medium. Mutant cells failed to form the asymmetric septum characteristic of sporulating cells and were defective in transcription of the earliest-expressed spo genes, that is, the genes dependent on the Spo0A phosphorelay. However, this early block in sporulation was partially overcome when cells of the citB mutant were induced to sporulate by resuspension in a poor medium. Accumulation of citrate in the mutant cells or in their culture fluid may be responsible for the early block, possibly because citrate can chelate divalent cations needed for the activity of the phosphorelay. PMID- 9393700 TI - A study of iterative type II polyketide synthases, using bacterial genes cloned from soil DNA: a means to access and use genes from uncultured microorganisms. AB - To examine as randomly as possible the role of the beta-ketoacyl and acyl carrier protein (ACP) components of bacterial type II polyketide synthases (PKSs), homologs of the chain-length-factor (CLF) genes were cloned from the environmental community of microorganisms. With PCR primers derived from conserved regions of known ketosynthase (KSalpha) and ACP genes specifying the formation of 16- to 24-carbon polyketides, two CLF (KSbeta) genes were cloned from unclassified streptomycetes isolated from the soil, and two were cloned from soil DNA without the prior isolation of the parent microorganism. The sequence and deduced product of each gene were distinct from those of known KSbeta genes and, by phylogenetic analysis, belonged to antibiotic-producing PKS gene clusters. Hybrid PKS gene cassettes were constructed with each novel KSbeta gene substituted for the actI-ORF2 or tcmL KSbeta subunit genes, along with the respective actI-ORF1 or tcmK KSalpha, tcmM ACP, and tcmN cyclase genes, and were found to produce an octaketide or decaketide product characteristic of the ones known to be made by the heterologous KSalpha gene partner. Since substantially less than 1% of the microorganisms present in soil are thought to be cultivatable by standard methods, this work demonstrates a potential way to gain access to a more extensive range of microbial molecular diversity and to biosynthetic pathways whose products can be tested for biological applications. PMID- 9393701 TI - An exo-poly-alpha-D-galacturonosidase, PehB, is required for wild-type virulence of Ralstonia solanacearum. AB - Ralstonia solanacearum, which causes bacterial wilt disease of many plant species, produces several extracellular plant cell wall-degrading enzymes that are suspected virulence factors. These include a previously described endopolygalacturonase (PG), PehA, and two exo-PGs. A gene encoding one of the exo PGs, pehB, was cloned from R. solanacearum K60. The DNA fragment specifying PehB contained a 2,103-bp open reading frame that encodes a protein of 74.2 kDa with a typical N-terminal signal sequence. The cloned pehB gene product cleaves polygalacturonic acid into digalacturonic acid units. The amino acid sequence of pehB resembles that of pehX, an exo-PG gene from Erwinia chrysanthemi, with 47.2% identity at the amino acid level. PehB also has limited similarity to plant exo PGs from Zea mays and Arabidopsis thaliana. The chromosomal pehB genes in R. solanacearum wild-type strain K60 and in an endo-PG PehA- strain were replaced with an insertionally inactivated copy of pehB. The resulting mutants were deficient in the production of PehB and of both PehA and PehB, respectively. The pehB mutant was significantly less virulent than the wild-type strain in eggplant virulence assays using a soil inoculation method. However, the pehA mutant was even less virulent, and the pehA pehB double mutant was the least virulent of all. These results suggest that PehB is required for a wild-type level of virulence in R. solanacearum although its individual role in wilt disease development may be minor. Together with endo-PG PehA, however, PehB contributes substantially to the virulence of R. solanacearum. PMID- 9393702 TI - Bacillus subtilis RNase III gene: cloning, function of the gene in Escherichia coli, and construction of Bacillus subtilis strains with altered rnc loci. AB - The rnc gene of Bacillus subtilis, which has 36% amino acid identity with the gene that encodes Escherichia coli RNase III endonuclease, was cloned in E. coli and shown by functional assays to encode B. subtilis RNase III (Bs-RNase III). The cloned B. subtilis rnc gene could complement an E. coli rnc strain that is deficient in rRNA processing, suggesting that Bs-RNase III is involved in rRNA processing in B. subtilis. Attempts to construct a B. subtilis rnc null mutant were unsuccessful, but a strain was constructed in which only a carboxy-terminal truncated version of Bs-RNase III was expressed. The truncated Bs-RNase III showed virtually no activity in vitro but was active in vivo. Analysis of expression of a copy of the rnc gene integrated at the amy locus and transcribed from a p(spac) promoter suggested that expression of the B. subtilis rnc is under regulatory control. PMID- 9393703 TI - Differential translocation of protein precursors across SecY-deficient membranes of Escherichia coli: SecY is not obligatorily required for translocation of certain secretory proteins in vitro. AB - SecY, a component of the protein translocation system in Escherichia coli, was depleted at a nonpermissive temperature in a strain which had a temperature sensitive polar effect on the expression of its secY. Membrane vesicles prepared from these cells, when grown at the nonpermissive temperature, contained about 5% SecY and similarly low levels of SecG. As expected, translocation of alkaline phosphatase precursors across these SecY-deficient membranes was severely impaired and appeared to be directly related to the decrease of SecY amounts. However, despite such a dramatic reduction in SecY and SecG levels, these membranes exhibited 50 to 70% of the wild-type translocation activity, including the processing of the signal peptide, of OmpA precursor (proOmpA). This translocation activity in SecY-deficient membranes was still SecA and ATP dependent and was not unique to proOmpA, as lipoprotein and lambda receptor protein precursors were also transported efficiently. Membranes that were reconstituted from these SecY-depleted membranes contained undetectable amounts of SecY yet were also shown to possess substantial translocation activity for proOmpA. These results indicate that the requirement of SecY for translocation is not obligatory for all secretory proteins and may depend on the nature of precursors. Consequently, it is unlikely that SecY is the essential core channel through which all precursors traverse across membranes; rather, SecY probably contributes to efficiency and specificity. PMID- 9393704 TI - Interaction of Bacillus subtilis purine repressor with DNA. AB - A purine repressor (PurR) mediates adenine nucleotide-dependent regulation of transcription initiation of the Bacillus subtilis pur operon. This repressor has been purified for the first time, and binding to control site DNA was characterized. PurR binds in vitro to four operons. Apparent Kd values for binding were 7 nM for the pur operon, 8 nM for purA, 13 nM for purR, and 44 nM for the pyr operon. In each case, DNase I footprints exhibited a pattern of protected and hypersensitive sites that extended over more than 60 bp. A GAAC-N24 GTTC sequence in the pur operon was necessary but not sufficient for the PurR-DNA interaction. However, this motif, which is conserved in the four binding sites, was not required for binding of PurR to purA. Thus, the common DNA recognition element for binding of PurR to the four operons is not known. Multiple PurR-pur operon DNA complexes having a binding stoichiometry that was either approximately two or six repressor molecules per DNA fragment were detected. The results of a torsional constraint experiment suggest that control site DNA forms one right handed turn around PurR. PMID- 9393705 TI - The arginine deiminase pathway in Rhizobium etli: DNA sequence analysis and functional study of the arcABC genes. AB - Sequence analysis upstream of the Rhizobium etli fixLJ homologous genes revealed the presence of three open reading frames homologous to the arcABC genes of Pseudomonas aeruginosa. The P. aeruginosa arcABC genes code for the enzymes of the arginine deiminase pathway: arginine deiminase, catabolic ornithine carbamoyltransferase (cOTCase), and carbamate kinase. OTCase activities were measured in free-living R. etli cells and in bacteroids isolated from bean nodules. OTCase activity in free-living cells was observed at a different pH optimum than OTCase activity in bacteroids, suggesting the presence of two enzymes with different characteristics and different expression patterns of the corresponding genes. The characteristics of the OTCase isolated from the bacteroids were studied in further detail and were shown to be similar to the properties of the cOTCase of P. aeruginosa. The enzyme has a pH optimum of 6.8 and a molecular mass of approximately 450 kDa, is characterized by a sigmoidal carbamoyl phosphate saturation curve, and exhibits a cooperativity for carbamoyl phosphate. R. etli arcA mutants, with polar effects on arcB and arcC, were constructed by insertion mutagenesis. Bean nodules induced by arcA mutants were still able to fix nitrogen but showed a significantly lower acetylene reduction activity than nodules induced by the wild type. No significant differences in nodule dry weight, plant dry weight, and number of nodules were found between the wild type and the mutants. Determination of the OTCase activity in extracts from bacteroids revealed a strong decrease in activity of this enzyme in the arcA mutant compared to the wild-type strain. Finally, we observed that expression of an R. etli arcA-gusA fusion was strongly induced under anaerobic conditions. PMID- 9393707 TI - Alkyl hydroperoxide reductase, catalase, MrgA, and superoxide dismutase are not involved in resistance of Bacillus subtilis spores to heat or oxidizing agents. AB - Only a single superoxide dismutase (SodA) was detected in Bacillus subtilis, and growing cells of a sodA mutant exhibited paraquat sensitivity as well as a growth defect and reduced survival at an elevated temperature. However, the sodA mutation had no effect on the heat or hydrogen peroxide resistance of wild-type spores or spores lacking the two major DNA protective alpha/beta-type small, acid soluble, spore proteins (termed alpha(-)beta(-) spores). Spores also had only a single catalase (KatX), as the two catalases found in growing cells (KatA and KatB) were absent. While a katA mutation greatly decreased the hydrogen peroxide resistance of growing cells, as found previously, katA, katB, and katX mutations had no effect on the heat or hydrogen peroxide resistance of wild-type or alpha( )beta(-) spores. Inactivation of the mrgA gene, which codes for a DNA-binding protein that can protect growing cells against hydrogen peroxide, also had no effect on spore hydrogen peroxide resistance. Inactivation of genes coding for alkyl hydroperoxide reductase, which has been shown to decrease growing cell resistance to alkyl hydroperoxides, had no effect on spore resistance to such compounds or on spore resistance to heat and hydrogen peroxide. However, Western blot analysis showed that at least one alkyl hydroperoxide reductase subunit was present in spores. Together these results indicate that proteins that play a role in the resistance of growing cells to oxidizing agents play no role in spore resistance. A likely reason for this lack of a protective role for spore enzymes is the inactivity of enzymes within the dormant spore. PMID- 9393706 TI - Fis, an accessorial factor for transcriptional activation of the mar (multiple antibiotic resistance) promoter of Escherichia coli in the presence of the activator MarA, SoxS, or Rob. AB - Transcription of the multiple antibiotic resistance marRAB operon increases when one of the sequence-related activators, MarA, SoxS, or Rob, binds to the "marbox" centered at -61.5 relative to the transcriptional start site. Previous deletion analyses showed that an adjacent upstream "accessory region" was needed to augment the marbox-dependent activation. To analyze the roles of the marbox and accessory regions on mar transcription, thirteen promoters, each with a different 5-bp transversion of the -96 to -32 sequence, were synthesized, fused to lacZ, and assayed for beta-galactosidase production in single-copy lysogens with appropriate genotypes. The accessory region is shown here to be a binding site for Fis centered at -81 and to bind Fis, a small DNA-binding and -bending protein, with a Kd of approximately 5 nM. The binding of MarA to the marbox and that of Fis to its site were independent of each other. MarA, SoxS, and Rob each activated the mar promoter 1.5-to 2-fold when it had a wild-type marbox but Fis was absent. In the presence of MarA, SoxS, or Rob, Fis further enhanced the activity of the promoter twofold provided the promoter was also capable of binding Fis. However, in the absence of MarA, SoxS, or Rob or in the absence of a wild-type marbox, Fis nonspecifically lowered the activity of the mar promoter about 25% whether or not a wild-type Fis site was present. Thus, Fis acts as an accessory transcriptional activator at the mar promoter. PMID- 9393708 TI - Physical and genetic map of the Clostridium acetobutylicum ATCC 824 chromosome. AB - A physical and genetic map of the Clostridium acetobutylicum ATCC 824 chromosome was constructed. The macrorestriction map for CeuI, EagI, and SstII was created by ordering the 38 restriction sites by one- and two-dimensional pulsed-field gel electrophoresis (PFGE) and by using an original strategy based on the CeuI enzyme and indirect end labelling by hybridization on both sides of the CeuI sites with rrs (16S RNA) and 3' rrl (23S RNA) probes. The circular chromosome was estimated to be 4.15 Mb in size, and the average resolution of the physical map is 110 kb. The chromosome contains 11 rrn loci, which are localized on 44% of the chromosome in a divergent transcriptional orientation regarding the presumed location of the replication origin. In addition to these 11 rrn operons, a total of 40 identified genes were mapped by hybridization experiments with genes from C. acetobutylicum and from various other clostridia as probes. The genetic map of C. acetobutylicum was compared to that of the three other endospore-forming bacteria characterized so far: Bacillus subtilis, Clostridium beijerinckii, and Clostridium perfringens. Parodoxically, the chromosomal backbone of C. acetobutylicum showed more similarity to that of B. subtilis than to those of the clostridia. PMID- 9393709 TI - Genetic requirements and mutational specificity of the Escherichia coli SOS mutator activity. AB - To better understand the mechanisms of SOS mutagenesis in the bacterium Escherichia coli, we have undertaken a genetic analysis of the SOS mutator activity. The SOS mutator activity results from constitutive expression of the SOS system in strains carrying a constitutively activated RecA protein (RecA730). We show that the SOS mutator activity is not enhanced in strains containing deficiencies in the uvrABC nucleotide excision-repair system or the xth and nfo base excision-repair systems. Further, recA730-induced errors are shown to be corrected by the MutHLS-dependent mismatch-repair system as efficiently as the corresponding errors in the rec+ background. These results suggest that the SOS mutator activity does not reflect mutagenesis at so-called cryptic lesions but instead represents an amplification of normally occurring DNA polymerase errors. Analysis of the base-pair-substitution mutations induced by recA730 in a mismatch repair-deficient background shows that both transition and transversion errors are amplified, although the effect is much larger for transversions than for transitions. Analysis of the mutator effect in various dnaE strains, including dnaE antimutators, as well as in proofreading-deficient dnaQ (mutD) strains suggests that in recA730 strains, two types of replication errors occur in parallel: (i) normal replication errors that are subject to both exonucleolytic proofreading and dnaE antimutator effects and (ii) recA730-specific errors that are not susceptible to either proofreading or dnaE antimutator effects. The combined data are consistent with a model suggesting that in recA730 cells error prone replication complexes are assembled at sites where DNA polymerization is temporarily stalled, most likely when a normal polymerase insertion error has created a poorly extendable terminal mismatch. The modified complex forces extension of the mismatch largely at the exclusion of proofreading and polymerase dissociation pathways. SOS mutagenesis targeted at replication-blocking DNA lesions likely proceeds in the same manner. PMID- 9393711 TI - Purification and properties of serine hydroxymethyltransferase from Sulfolobus solfataricus. AB - Serine hydroxymethyltransferase (SHMT) catalyzes the reversible cleavage of serine to glycine with the transfer of the one-carbon group to tetrahydrofolate to form 5,10-methylenetetrahydrofolate. No SHMT has been purified from a nonmethanogenic Archaea strain, in part because this group of organisms uses modified folates as the one-carbon acceptor. These modified folates are not readily available for use in assays for SHMT activity. This report describes the purification and characterization of SHMT from the thermophilic organism Sulfolobus solfataricus. The exchange of the alpha-proton of glycine with solvent protons in the absence of the modified folate was used as the activity assay. The purified protein catalyzes the synthesis of serine from glycine and a synthetic derivative of a fragment of the natural modified folate found in S. solfataricus. Replacement of the modified folate with tetrahydrofolate did not support serine synthesis. In addition, this SHMT also catalyzed the cleavage of both allo threonine and beta-phenylserine in the absence of the modified folate. The cleavage of these two amino acids in the absence of tetrahydrofolate is a property of other characterized SHMTs. The enzyme contains covalently bound pyridoxal phosphate. Sequences of three peptides showed significant similarity with those of peptides of SHMTs from two methanogens. PMID- 9393712 TI - Purification and biochemical characterization of a hydroxyneurosporene desaturase involved in the biosynthetic pathway of the carotenoid spheroidene in Rhodobacter sphaeroides. AB - Hydroxyneurosporene desaturase is involved in the carotenoid biosynthetic pathway of Rhodobacter species. The gene encoding this enzyme was expressed in Escherichia coli, purified, and biochemically characterized. The resulting protein contained an N-terminal six-histidine extension which derived from the cloning vector; this allowed for a one-step purification of the enzyme to homogeneity after solubilization with Nonidet P-40. The hydrogen acceptor in the C-3,4 desaturation reaction was molecular oxygen. NAD+, NADP+, and flavin adenine dinucleotide had no influence on enzymatic activity. Different acyclic 1 hydroxycarotenoids were tested as substrates. Very good conversion was achieved with 1-hydroxyneurosporene and 1-hydroxylycopene, whereas 1-hydroxy-gamma carotene and 1,1'-dihydroxylycopene were much less effective. From 1'-hydroxy-3,4 didehydrolycopene only trace amounts of product were obtained, and 1 methoxyneurosporene was not converted by purified hydroxyneurosporene desaturase. A Km of 13.4 microM was determined for 1-hydroxyneurosporene. PMID- 9393710 TI - NtrC is required for control of Klebsiella pneumoniae NifL activity. AB - In response to molecular oxygen and/or fixed nitrogen, the product of the Klebsiella pneumoniae nitrogen fixation L (nifL) gene inhibits NifA-mediated transcriptional activation. Nitrogen regulation of NifL function occurs at two levels: transcription of the nifLA operon is regulated by the general Ntr system, and the activity of NifL is controlled by an unknown mechanism. We have studied the regulation of NifL activity in Escherichia coli and Salmonella typhimurium by monitoring its inhibition of NifA-mediated expression of a K. pneumoniae phi(nifH'-'lacZ) fusion. The activity of the NifL protein transcribed from the tac promoter is regulated well in response to changes of oxygen and/or nitrogen status, indicating that no nif- or K. pneumoniae-specific product is required. Unexpectedly, strains carrying ntrC (glnG) null alleles failed to release NifL inhibition, despite the fact that synthesis of NifL was no longer under Ntr control. Additional evidence indicated that it is indeed the transcriptional activation capacity of NtrC, rather than its repression capacity, that is needed, and hence it is a plausible hypothesis that NtrC activates transcription of a gene(s) whose product(s) in turn functions to relieve NifL inhibition under nitrogen-limiting conditions. PMID- 9393713 TI - Cloning and genetic analysis of the UV resistance determinant (uvr) encoded on the Enterococcus faecalis pheromone-responsive conjugative plasmid pAD1. AB - The conjugative pheromone-responsive plasmid pAD1 (59.6 kb) of Enterococcus faecalis encodes a UV resistance determinant (uvr) in addition to the hemolysin bacteriocin determinant. pAD1 enhances the UV resistance of wild-type E. faecalis FA2-2 and E. faecalis UV202, which is a UV-sensitive derivative of E. faecalis JH2-2. A 2.972-kb fragment cloned from between 27.7 and 30.6 kb of the pAD1 map conferred UV resistance function on UV202. Sequence analysis showed that the cloned fragment contained three open reading frames designated uvrA, uvrB, and uvrC. The uvrA gene is located on the pAD1 map between 28.1 and 29.4 kb. uvrB is located between 30.1 and 30.3 kb, and uvrC is located between 30.4 and 30.6 kb on the pAD1 map. The uvrA, uvrB, and uvrC genes encode sequences of 442, 60, and 74 amino acids, respectively. The deduced amino acid sequence of the uvrA-encoded protein showed 20% homology of the identical residues with the E. coli UmuC protein. Tn917 insertion mutagenesis and deletion mutant analysis of the cloned fragment showed that uvrA conferred UV resistance. A palindromic sequence, 5' GAACNGTTC-3', which is identical to the consensus sequence found within the putative promoter region of the Bacillus subtilis DNA damage-inducible genes, was located within the promoter region of uvrA. Two uvrA transcripts of different lengths (i.e., 1.54 and 2.14 kb) which terminate at different points downstream of uvrA were detected in UV202 carrying the deletion mutant containing uvrA. The longer transcript, 2.14 kb, was not detected in UV202 carrying the deletion mutant containing both uvrA and uvrB, which suggests that uvrB encodes a terminator for the uvrA transcript. The uvrA transcript was not detected in any significant quantity in UV202 carrying the cloned fragment containing uvrA, uvrB, and uvrC; on the other hand, the 1.54-kb uvrA transcript was detected in the strain exposed to mitomycin C, which suggests that the UvrC protein functions as a regulator of uvrA. PMID- 9393715 TI - Structure and evolution of NGRRS-1, a complex, repeated element in the genome of Rhizobium sp. strain NGR234. AB - Much of the remarkable ability of Rhizobium sp. strain NGR234 to nodulate at least 110 genera of legumes, as well as the nonlegume Parasponia andersonii, stems from the more than 80 different Nod factors it secretes. Except for nodE, nodG, and nodPQ, which are on the chromosome, most Nod factor biosynthesis genes are dispersed over the 536,165-bp symbiotic plasmid, pNGR234a. Mosaic sequences and insertion sequences (ISs) comprise 18% of pNGR234a. Many of them are clustered, and these IS islands divide the replicon into large blocks of functionally related genes. At 6 kb, NGRRS-1 is a striking example: there is one copy on pNGR234a and three others on the chromosome. DNA sequence comparisons of two NGRRS-1 elements identified three types of IS, NGRIS-2, NGRIS-4, and NGRIS 10. Here we show that all four copies of NGRRS-1 probably originated from transposition of NGRIS-4 into a more ancient IS-like sequence, NGRIS-10. Remarkably, all nine copies of NGRIS-4 have transposed into other ISs. It is unclear whether the accumulation of potentially mutagenic sequences in large clusters is due to the nature of the IS involved or to some selection process. Nevertheless, a direct consequence of the preferential targeting of transposons into such IS islands is to minimize the likelihood of disrupting vital functions. PMID- 9393714 TI - Negative regulation of mutS and mutH repair gene expression by the Hfq and RpoS global regulators of Escherichia coli K-12. AB - The MutS, MutL, and MutH proteins play major roles in several DNA repair pathways. We previously reported that the cellular amounts of MutS and MutH decreased by as much as 10-fold in stationary-phase cultures. Consequently, we tested whether the amounts of MutS, MutL, and MutH were regulated by two global regulators, RpoS (sigma38) and Hfq (HF-I [putative RNA chaperone]), which are involved in stationary-phase transition. We report here that mutations in hfq and rpoS reversed the stationary-phase down-regulation of the amounts of MutS and MutH. hfq regulation of the amount of MutS in stationary-phase cultures was mediated by RpoS-dependent and -independent mechanisms, whereas hfq regulation of the amount of MutH was mediated only through RpoS. Consistent with this interpretation, the amount of MutS but not MutH was regulated by Hfq, but not RpoS, in exponentially growing cells. The amount of MutL remained unchanged in rpoS, hfq-1, and rpoS+, hfq+ strains in exponentially growing and stationary phase cultures and served as a control. The beta-galactosidase activities of single-copy mutS-lacZ operon and gene fusions suggested that hfq regulates mutS posttranscriptionally in exponentially growing cultures. RNase T2 protection assays revealed increased amounts of mutS transcript that are attributed to increased mutS transcript stability in hfq-1 mutants. Lack of Hfq also increased the amounts and stabilities of transcripts initiated from P(miaA) and P1hfqHS, two of the promoters for hfq, suggesting autoregulation, but did not change the half-life of bulk mRNA. These results suggest that the amounts of MutS and MutH may be adjusted in cells subjected to different stress conditions by an RpoS dependent mechanism. In addition, Hfq directly or indirectly regulates several genes, including mutS, hfq, and miaA, by an RpoS-independent mechanism that destabilizes transcripts. PMID- 9393716 TI - The 2microm-plasmid-encoded Rep1 and Rep2 proteins interact with each other and colocalize to the Saccharomyces cerevisiae nucleus. AB - The efficient partitioning of the 2microm plasmid of Saccharomyces cerevisiae at cell division requires two plasmid-encoded proteins (Rep1p and Rep2p) and a cis acting locus, REP3 (STB). By using protein hybrids containing fusions of the Rep proteins to green fluorescent protein (GFP), we show here that fluorescence from GFP-Rep1p or GFP-Rep2p is almost exclusively localized in the nucleus in a cir+ strain. Nuclear localization of GFP-Rep1p and GFP-Rep2p, though discernible, is less efficient in a cir(0) host. GFP-Rep2p or GFP-Rep1p is able to promote the stability of a 2microm circle-derived plasmid harboring REP1 or REP2, respectively, in a cir(0) background. Under these conditions, fluorescence from GFP-Rep2p or GFP-Rep1p is concentrated within the nucleus, as is the case in cir+ cells. This characteristic nuclear accumulation is not dependent on the expression of the FLP or RAF1 gene of the 2microm circle. Nuclear colocalization of Rep1p and Rep2p is consistent with the hypothesis that the two proteins directly or indirectly interact to form a functional bipartite or high-order protein complex. Immunoprecipitation experiments as well as baiting assays using GST-Rep hybrid proteins suggest a direct interaction between Rep1p and Rep2p which, in principle, may be modulated by other yeast proteins. Furthermore, these assays provide evidence for Rep1p-Rep1p and Rep2p-Rep2p associations as well. The sum of these interactions may be important in controlling the effective cellular concentration of the Rep1p-Rep2p complex. PMID- 9393717 TI - Escherichia coli cells expressing a mutant glyV (glycine tRNA) gene have a UVM constitutive phenotype: implications for mechanisms underlying the mutA or mutC mutator effect. AB - Transfection of M13 single-stranded viral DNA bearing a 3,N4-ethenocytosine lesion into Escherichia coli cells pretreated with UV results in a significant elevation of mutagenesis at the lesion site compared to that observed in untreated cells. This response, termed UVM, for UV modulation of mutagenesis, is induced by a variety of DNA-damaging agents and is distinct from known cellular responses to DNA damage, including the SOS response. This report describes our observation, as a part of our investigation of the UVM phenomenon, that E. coli cells bearing a mutA or mutC allele display a UVM-constitutive phenotype. These mutator alleles were recently mapped (M. M. Slupska, C. Baikalov, R. Lloyd, and J. H. Miller, Proc. Natl. Acad. Sci. USA 93:4380-4385, 1996) to the glyV (mutA) and glyW (mutC) tRNA genes. Each mutant allele was shown to arise by an identical mutation in the anticodon sequence such that the mutant tRNAs could, in principle, mistranslate aspartate codons in mRNA as glycine at a low level. Because a UVM-constitutive phenotype resulting from a mutation in a tRNA gene was unexpected, we undertook a series of experiments designed to test whether the phenotype was indeed mediated by the expression of mutant glycine tRNAs. We placed either a wild-type or a mutant glyV gene under the control of a heterologous inducible promoter on a plasmid vector. E. coli cells expressing the mutant glyV gene displayed all three of the following phenotypes: (i) missense suppression of a test allele, (ii) a mutator phenotype measured by mutation to rifampin resistance, and (iii) a UVM-constitutive phenotype. These phenotypes were not associated with cells expressing the wild-type glyV gene or with cells in which the mutant allele was present but was not transcriptionally induced. These observations provide strong support for the idea that expression of mutant tRNA can confer a mutator phenotype, including the UVM-constitutive phenotype observed in mutA and mutC cells. However, our data imply that low-level mistranslation of the epsilon subunit of polymerase III probably does not account for the observed UVM-constitutive phenotype. Our results also indicate that mutA deltarecA double mutants display a normal UVM phenotype, suggesting that the mutA effect is recA dependent. The observations reported here raise a number of intriguing questions and raise the possibility that the UVM response is mediated through transient alteration of the replication environment. PMID- 9393718 TI - Identification and characterization of the niddamycin polyketide synthase genes from Streptomyces caelestis. AB - The genes encoding the polyketide synthase (PKS) portion of the niddamycin biosynthetic pathway were isolated from a library of Streptomyces caelestis NRRL 2821 chromosomal DNA. Analysis of 40 kb of DNA revealed the presence of five large open reading frames (ORFs) encoding the seven modular sets of enzymatic activities required for the synthesis of a 16-membered lactone ring. The enzymatic motifs identified within each module were consistent with those predicted from the structure of niddamycin. Disruption of the second ORF of the PKS coding region eliminated niddamycin production, demonstrating that the cloned genes are involved in the biosynthesis of this compound. PMID- 9393719 TI - Nested DNA inversion of Campylobacter fetus S-layer genes is recA dependent. AB - Wild-type strains of Campylobacter fetus are covered by a monomolecular array of surface layer proteins (SLPs) critical for virulence. Each cell possesses eight SLP gene cassettes, tightly clustered in the genome, that encode SLPs of 97 to 149 kDa. Variation of SLP expression occurs by a mechanism of nested DNA rearrangement that involves the inversion of a 6.2-kb sapA promoter-containing element alone or together with one or more flanking SLP gene cassettes. The presence of extensive regions of identity flanking the 5' and 3' ends of each SLP gene cassette and of a Chi-like recognition sequence within the 5' region of identity suggests that rearrangement of SLP gene cassettes may occur by a generalized (RecA-dependent) homologous recombination pathway. To explore this possibility, we cloned C. fetus recA and created mutant strains by marker rescue, in which recA is disrupted in either S+ or S- strains. These mutants then were assessed for their abilities to alter SLP expression either in the presence or absence of a complementary shuttle plasmid harboring native recA. In contrast to all previously reported programmed DNA inversion systems, inversion in C. fetus is recA dependent. PMID- 9393721 TI - Methylmalonyl coenzyme A selectivity of cloned and expressed acyltransferase and beta-ketoacyl synthase domains of mycocerosic acid synthase from Mycobacterium bovis BCG. AB - Methyl-branched fatty acids and polyketides occur in a variety of living organisms. Previous studies have established that multifunctional enzymes use methylmalonyl coenzyme A (CoA) as the substrate to generate methyl-branched products such as mycocerosic acids and polyketides. However, we do not know which of the component activities show selectivity for methylmalonyl-CoA in any biological system. A comparison of homologies of the domains of the multifunctional synthases that selectively use malonyl-CoA or methylmalonyl-CoA suggested that the acyltransferase (AT) and beta-ketoacyl synthase (KS) domains might be responsible for the substrate selectivity. To test this hypothesis, we expressed the AT and KS domains of the mycocerosic acid synthase (MAS) gene from Mycobacterium bovis BCG in Escherichia coli and examined whether they confer to synthases that normally do not use methylmalonyl-CoA the ability to incorporate methylmalonyl-CoA into fatty acids. Both the AT and the KS domains of MAS showed selectivity for methylmalonyl-CoA over malonyl-CoA. Acyl carrier protein (ACP) dependent elongation of the n-C12 acyl primer mainly by one methylmalonyl-CoA unit was catalyzed by an E. coli fatty acid synthase preparation only in the presence of the expressed MAS domains. An ACP-dependent elongation of the n-C20 acyl primer by one methylmalonyl-CoA extender unit was catalyzed by fatty acid synthase from Mycobacterium smegmatis only in the presence of the expressed MAS domains. These results show methylmalonyl-CoA selectivity for the AT and KS domains of MAS. These domains may be useful in producing novel polyketides by genetic engineering. PMID- 9393720 TI - A quorum-sensing system in the free-living photosynthetic bacterium Rhodobacter sphaeroides. AB - Rhodobacter sphaeroides is a free-living, photoheterotrophic bacterium known for its genomic and metabolic complexity. We have discovered that this purple photosynthetic organism possesses a quorum-sensing system. Quorum sensing occurs in a number of eukaryotic host-associated gram-negative bacteria. In these bacteria there are two genes required for quorum sensing, the luxR and luxI homologs, and there is an acylhomoserine lactone signal molecule synthesized by the product of the luxI homolog. In R. sphaeroides, synthesis of a novel homoserine lactone signal, 7,8-cis-N-(tetradecenoyl)homoserine lactone, is directed by a luxI homolog termed cerI. Two open reading frames immediately upstream of cerI are proposed to be components of the quorum-sensing system. The first of these is a luxR homolog termed cerR, and the second is a small open reading frame of 159 bp. Inactivation of cerI in R. sphaeroides results in mucoid colony formation on agar and formation of large aggregates of cells in liquid cultures. Clumping of CerI mutants in liquid culture is reversible upon addition of the acylhomoserine lactone signal and represents a phenotype unlike those controlled by quorum sensing in other bacteria. PMID- 9393723 TI - Terminal inverted repeats of insertion sequence IS30 serve as targets for transposition. AB - In the present study, we demonstrate that the terminal inverted repeats of the Escherichia coli insertion sequence IS30 are functional target sites for the transposition of the (IS30)2 dimer, which represents an intermediate structure in the transposition of IS30. Comparative analysis of various target regions revealed that the left and right ends differ in their "attractivity." In our experiments, the joined left and right ends, i.e., the (IS30)2 intermediate structure, was found to be the most preferred target. It was also shown that flanking sequences can influence the target activity of the terminal repeats. The functional part of the target region was localized in the inverted repeats by means of mutational analysis, and it corresponds to the binding site of IS30 transposase. Insertion of 1 bp into the right inverted repeat resulted in unusual target duplication accompanied by gene conversion. The choice of the terminal inverted repeats as targets in transposition leads to the reconstruction of the (IS30)2 structure, which may induce a cascade of further rearrangements. Therefore, this process can play a role in the evolution of the genome. PMID- 9393722 TI - Conserved motifs II to VI of DNA helicase II from Escherichia coli are all required for biological activity. AB - There are seven conserved motifs (IA, IB, and II to VI) in DNA helicase II of Escherichia coli that have high homology among a large family of proteins involved in DNA metabolism. To address the functional importance of motifs II to VI, we employed site-directed mutagenesis to replace the charged amino acid residues in each motif with alanines. Cells carrying these mutant alleles exhibited higher UV and methyl methanesulfonate sensitivity, increased rates of spontaneous mutagenesis, and elevated levels of homologous recombination, indicating defects in both the excision repair and mismatch repair pathways. In addition, we also changed the highly conserved tyrosine(600) in motif VI to phenylalanine (uvrD309, Y600F). This mutant displayed a moderate increase in UV sensitivity but a decrease in spontaneous mutation rate, suggesting that DNA helicase II may have different functions in the two DNA repair pathways. Furthermore, a mutation in domain IV (uvrD307, R284A) significantly reduced the viability of some E. coli K-12 strains at 30 degrees C but not at 37 degrees C. The implications of these observations are discussed. PMID- 9393724 TI - Characterization of the acc operon from the nopaline-type Ti plasmid pTiC58, which encodes utilization of agrocinopines A and B and susceptibility to agrocin 84. AB - The acc locus from the Ti plasmid pTiC58 confers utilization of and chemotaxis toward agrocinopines A and B (A+B), as well as susceptibility to a highly specific antiagrobacterial antibiotic, agrocin 84. DNA sequence analyses revealed that acc is composed of eight open reading frames, accR and accA through accG. Previous work showed that accR encodes the repressor which regulates this locus, and accA codes for the periplasmic binding protein of the agrocinopine transport system (S. Beck Von Bodman, G. T. Hayman, and S. K. Farrand, Proc. Natl. Acad. Sci. USA 89:643-647, 1992; G. T. Hayman, S. Beck Von Bodman, H. Kim, P. Jiang, and S. K. Farrand, J. Bacteriol. 175:5575-5584, 1993). The predicted proteins from accA through accE, as a group, have homology to proteins that belong to the ABC-type transport system superfamily. The predicted product of accF is related to UgpQ of Escherichia coli, which is a glycerophosphoryl diester phosphodiesterase, and also to agrocinopine synthase coded for by acs located on the T-DNA. The translated product of accG is related to myoinositol 1 (or 4) monophosphatases from various eucaryotes. Analyses of insertion mutations showed that accA through accE are required for transport of both agrocin 84 and agrocinopines A+B, while accF and accG are required for utilization of the opines as the sole source of carbon. Mutations in accF or accG did not abolish transport of agrocin 84, although we observed slower removal of the antibiotic from the medium by the accF mutant compared to the wild type. However, the insertion mutation in accF abolished detectable uptake of agrocinopines A+B. A mutation in accG had no effect on transport of the opines. The accF mutant was not susceptible to agrocin 84 although it took up the antibiotic. This finding suggests that agrocin 84 is activated by AccF after being transported into the bacterial cell. PMID- 9393725 TI - Specificities of FemA and FemB for different glycine residues: FemB cannot substitute for FemA in staphylococcal peptidoglycan pentaglycine side chain formation. AB - The femAB operon codes for two nearly identical approximately 50-kDa proteins involved in the formation of the staphylococcal pentaglycine interpeptide bridge. Sequencing and analysis of the femA region of mutants isolated by chemical mutagenesis and selection for lysostaphin resistance revealed point mutations leading to the expression of truncated FemA proteins. These femA mutants, although still producing an intact FemB, exhibited a phenotype identical as that described for femAB double mutants. Thus, FemA seems to be essential for the addition of glycine residues 2 and 3 only, whereas FemB is involved in the attachment of exclusively glycine residues 4 and 5. Although FemB has 39% identity with FemA, it cannot substitute for FemA. The FemA and FemB proteins seem to be highly specific in regard to the position of the glycine residues that they attach. PMID- 9393726 TI - Essential role of the consensus nucleotide-binding site of PtlH in secretion of pertussis toxin from Bordetella pertussis. AB - PtlH is a member of a specialized set of transport proteins that is essential for secretion of pertussis toxin (PT) from Bordetella pertussis. Previously, PtlH was shown to contain a consensus nucleotide-binding motif. Here, we demonstrate that introduction of plasmids containing mutant forms of ptlH, altered in the putative nucleotide-binding region, into a wild-type strain of B. pertussis resulted in inhibition of PT secretion. Thus, this region of PtlH appears to be essential for protein function. Moreover, the observed dominant negative phenotype suggests that PtlH either functions as a multimer or interacts with another component necessary for secretion of PT. PMID- 9393727 TI - A gene (wbbL) from Serratia marcescens N28b (O4) complements the rfb-50 mutation of Escherichia coli K-12 derivatives. AB - A cosmid-based genomic library of Serratia marcescens N28b was introduced into Escherichia coli DH5alpha, and clones were screened for serum resistance. One clone was found resistant to serum, to bacteriocin 28b, and to bacteriophages TuIa and TuIb. This clone also showed O antigen in its lipopolysaccharide. Subcloning and sequencing experiments showed that a 2,124-bp DNA fragment containing the rmlD and wbbL genes was responsible for the observed phenotypes. On the basis of amino acid similarity, we suggest that the 288-residue RmlD protein is a dTDP-L-rhamnose synthase. Plasmid pJT102, containing only the wbbL gene, was able to induce O16-antigen production and serum resistance in E. coli DH5alpha. These results suggest that the 282-residue WbbL protein is a rhamnosyltransferase able to complement the rJb-50 mutation in E. coli K-12 derivatives, despite the low level of amino acid identity between WbbL and the E. coli rhamnosyltransferase (24.80%). S. marcescens N28b rmlD and wbbL mutants were constructed by mobilization of suicide plasmids containing a portion of rmlD or wbbL. These insertion mutants were unable to produce O antigen; since strain N28b produces O4 antigen, these results suggest that both genes are involved in O4 antigen biosynthesis. PMID- 9393728 TI - Role of NifS in maturation of glutamine phosphoribosylpyrophosphate amidotransferase. AB - Glutamine phosphoribosylpyrophosphate amidotransferase from Bacillus subtilis is synthesized as an inactive precursor that requires two maturation steps: incorporation of a [4Fe-4S] center and cleavage of an 11-residue NH2-terminal propeptide. Overproduction from a multicopy plasmid in Escherichia coli leads to the formation of soluble proenzyme and mature enzyme forms as well as a small fraction of insoluble proenzyme. Heterologous expression of Azotobacter vinelandii nifS from a compatible plasmid increased the maturation of the soluble proenzyme three- to fourfold without influencing the content of the insoluble fraction. These results support a role for NifS in heterologous Fe-S cluster assembly and enzyme maturation. PMID- 9393729 TI - The rpoN (sigma54) gene from Listeria monocytogenes is involved in resistance to mesentericin Y105, an antibacterial peptide from Leuconostoc mesenteroides. AB - To gain insight into the mode of action of mesentericin Y105, a bacteriocin bactericidal agent against Listeria monocytogenes, we undertook to identify the listerial factors mediating this susceptibility by using a genetic approach. Transposon mutants resistant to the bacteriocin were obtained. One of them corresponded to a transposon insertion in a gene (rpoN) encoding a putative protein (447 amino acids) with strong homologies to alternative transcriptional sigma54 factors, including that of Bacillus subtilis (38% identity). Complementation experiments with the wild-type rpoN gene demonstrated that the insertion in rpoN was responsible for the resistance phenotype in L. monocytogenes. Moreover, expression of the L. monocytogenes rpoN gene in an rpoN mutant strain of B. subtilis promoted transcription of a sigma54-dependent operon in the presence of the associated regulator. These results demonstrate that the L. monocytogenes rpoN gene encodes a new sigma54 factor. PMID- 9393730 TI - Proteins induced in Escherichia coli by benzoic acid. AB - Proteins induced by benzoic acid in Escherichia coli were observed on two dimensional electrophoretic gels (2-D gels). Cultures were grown in glucose-rich medium in the presence or absence of 20 mM benzoate at an external pH of 6.5, where the pH gradient (deltapH) is large and benzoate accumulates, and at an external pH of 8.0, where deltapH is inverted and little benzoate is taken up. Radiolabeled proteins were separated on 2-D gels and were identified on the basis of the index of VanBogelen and Neidhardt. In the absence of benzoic acid, little difference was seen between pH 6.5 and pH 8.0; this confirms that the mechanisms of protein homeostasis in this range are constitutive, including the transition between positive and inverted deltapH. Addition of benzoate at pH 6.5 increased the expression of 33 proteins. Twelve of the benzoate-induced proteins were induced at pH 8.0 as well, and nine of these matched proteins induced by the uncoupler dinitrophenol. Eighteen proteins were induced by benzoate only at pH 6.5, not at pH 8.0, and were not induced by dinitrophenol. One may be the iron and pH regulator Fur, which regulates acid tolerance in Salmonella spp. The other 13 proteins had not been identified previously. The proteins induced by benzoate only at a low pH may reflect responses to internal acidification or to accumulation of benzoate. PMID- 9393731 TI - Mutational analysis of amiloride sensitivity of the NhaA Na+/H+ antiporter from Vibrio parahaemolyticus. AB - The activity of the NhaA Na+/H+ antiporter of Vibrio parahaemolyticus is inhibited by amiloride. We found an amino acid sequence in the NhaA that was identical to a putative amiloride binding domain of the Na+/H+ exchanger in mammalian cells. We constructed mutant NhaAs that had amino acid substitutions in the putative amiloride binding domain by site-directed mutagenesis. These include V62L (Val62 replaced by Leu), F63Y, F64Y, and L65F. Most mutant NhaAs showed decreased sensitivity for amiloride. Among these, the F64Y mutant NhaA showed the least amiloride sensitivity, with a Ki value 7 to 10 times greater than that in the wild type. Thus, the sequence between residues V62 and L65 in NhaA, especially F64, is very important for the inhibitory effect of amiloride on the antiporter. PMID- 9393732 TI - Contribution of glucose kinase to glucose repression of xylose utilization in Bacillus megaterium. AB - The glk gene from Bacillus megaterium, which encodes glucose kinase, was isolated and analyzed. Disruption by a transcriptional glk-luxAB fusion indicated that glk is the only glucose kinase gene in that strain but did not affect growth of that mutant on glucose. Determination of luciferase activity under various growth conditions revealed constitutive transcription of glk. Expression of a xylA-lacZ fusion was repressed by glucose in the strain with the glk disruption about twofold less efficiently than in the wild type. The potential contribution of glk expression to glucose repression is discussed. PMID- 9393733 TI - Functional domains of a zinc metalloprotease from Vibrio vulnificus. AB - Vibrio vulnificus, an opportunistic human pathogen causing wound infection and septicemia, secretes a 45-kDa metalloprotease (V. vulnificus protease; VVP). A plasmid which carries the entire vvp gene subcloned into pBluescriptIIKS+ was transformed into Escherichia coli DH5alpha for overproduction of the protease. The 45-kDa recombinant protease (rVVP) was isolated from the periplasmic fraction of the transformant by ammonium sulfate precipitation followed by column chromatography on phenyl Sepharose. Biochemical characterization of the isolated rVVP showed that the recombinant protease was identical to that produced by V. vulnificus. When rVVP was incubated at 37 degrees C, a 35-kDa fragment was generated through autoproteolytic removal of the C-terminal peptide. This 35-kDa fragment (rVVP-N) was found to have sufficient proteolytic activity toward oligopeptides and soluble proteins but had markedly reduced activity toward insoluble proteins. Lineweaver-Burk plot analysis indicated increased Km values of rVVP-N for all of the protein substrates. rVVP, but not rVVP-N, was shown to agglutinate rabbit erythrocytes, bind to the erythrocyte ghosts, and digest the ghost membrane proteins. These results strongly suggest that rVVP (and VVP) consists of at least two functional domains: an N-terminal 35-kDa polypeptide mediating proteolysis and a C-terminal 10-kDa polypeptide which may be essential for efficient attachment to protein substrates and erythrocyte membranes. PMID- 9393734 TI - Carcinoma-associated expression of core 2 beta-1,6-N acetylglucosaminyltransferase gene in human colorectal cancer: role of O-glycans in tumor progression. AB - Recently, it was demonstrated that an increased level of NeuNAc alpha2-3Gal beta1 4(Fuc alpha1-3)GlcNAc beta-R (sialyl Le(x)) and NeuNAc alpha2-3Gal beta1-3(Fuc alpha1-4)GlcNAc beta-R (sialyl Le(a)) expression on the surface of colorectal cancer cells is positively correlated with progression of the disease. It has not been determined, however, which type of glycans, N- or O-glycans, is more closely associated with progression when cancer cells express those oligosaccharides. To address this problem, we have examined expression of sialyl Le(a) and sialyl Le(x), those oligosaccharides in O-glycans, and core 2 beta-1,6-N acetylglucosaminyltransferase (C2GnT) transcripts in colorectal cancer specimens from 46 patients and compared those results with clinicopathological variables. C2GnT is a glycosyltransferase that is responsible for the core 2 branch, which is critical for biosynthesis of sialyl Le(a) and sialyl Le(x) in O-glycans. Sialyl Le(a) and sialyl Le(x) were determined by immunohistochemistry, and C2GnT transcripts were detected by reverse transcription-PCR. Sialyl Le(a) or sialyl Le(x) in O-glycans was assessed by combining immunohistochemistry for sialyl Le(a) or sialyl Le(x) with reverse transcription-PCR for C2GnT. Sialyl Le(a), detected on cancer cells in 74% of patients, was well correlated with lymph node metastasis, whereas sialyl Le(a) and sialyl Le(x) in O-glycans, which were specifically detected in cancer tissues of 50 and 61% of patients, respectively, were closely associated with lymphatic and venous invasion. In addition, C2GnT, which was specifically detected in cancer tissues of 63% of patients, was closely correlated with the vessel invasion, as well as depth of tumor invasion. These results strongly suggest that sialyl Le(a) and sialyl Le(x) in O-glycans and C2GnT, expressed in cancer cells, may play important roles in tumor progression through vessel or direct invasion. PMID- 9393735 TI - Absence of Fhit protein in primary lung tumors and cell lines with FHIT gene abnormalities. AB - Genomic alterations and abnormal expression of the FHIT gene at 3p14.2 have been observed in cell lines and primary tumors of the lung. To correlate FHIT locus DNA and RNA lesions with effects on Fhit protein expression, we have analyzed 11 lung cancer cell lines, 15 small cell lung carcinomas, and 38 pairs of non-small cell primary tumors and bronchial mucosa specimens by molecular genetic and immunocytochemical methods. Using specific antibodies against the Fhit protein, we observed concordance between RNA abnormalities and lack of Fhit protein expression in lung tumors and cell lines. In addition, absence of Fhit protein in some precancerous dysplastic lesions suggested that FHIT inactivation may occur at an early phase of lung carcinogenesis. PMID- 9393736 TI - Patterns of chromosomal alterations in metastasizing and nonmetastasizing primary head and neck carcinomas. AB - In an attempt to define chromosomal alterations that are associated with the metastatic phenotype, we investigated a total of 29 metastasizing (pN+) and 19 non-metastasizing (pN0) head and neck squamous cell carcinomas by comparative genomic hybridization (CGH). The analysis indicated that the pN0 tumors carried preferentially overrepresentations of chromosomes 5p, 6p, and 7p and that the pN+ tumors were frequently characterized by deletions on chromosomes 7q, 10q, 11p, 11q, 15q, and 20p and overrepresentations of the chromosomes 19q and 20q. In particular, the use of difference histograms and statistical analysis indicated that the deletions on chromosomes 10q25-q26 and 11p13-p14 were highly significant for metastasizing carcinomas. The findings on chromosome 10q were supported by loss of heterozygosity analysis in the primary tumors and eight synchronous lymph node metastases using four microsatellite polymorphisms. The data suggest that distinct patterns of genetic lesions are responsible for the metastatic phenotype of head and neck squamous cell carcinomas. PMID- 9393737 TI - Intracellular localization of p53 tumor suppressor protein in gamma-irradiated cells is cell cycle regulated and determined by the nucleus. AB - DNA damage leads to the stabilization of p53 protein and its translocation to the nucleus, resulting in activation or suppression of p53-responsive genes. However, a significant proportion of cell nuclei remain negative for p53 and p53-inducible cyclin-dependent kinase inhibitor p21waf1 after a single dose of gamma irradiation. Quantitation of DNA content in p53-positive and -negative nuclei 4-6 h after 10 Gy of gamma-irradiation of human breast carcinoma MCF7 cells, fibrosarcoma HT1080 cells, and diploid skin fibroblasts showed that p53 and p21waf1 nuclear accumulation occurs predominantly in the G1 phase and at the beginning of the S phase of the cell cycle. The majority of the nuclei in late S phase and in G2-M phase remained p53- and p21waf1-negative. This suggests that there is a cell cycle window during which p53 can accumulate in the nucleus and activate expression of p21waf1. To determine whether cell cycle-dependent distribution of p53 is caused by cytoplasmic modifications of p53 protein or by properties of the nucleus, p53 localization was analyzed in multinucleated cells obtained by polyethylene glycol-mediated cell fusion. Dramatic differences in p53 accumulation were found among the nuclei in individual multinucleated cells. Distribution of p53-positive and -negative nuclei among the phases of the cell cycle was similar to that observed in a regular cell population. These results suggest that the observed differences in p53 accumulation in the nuclei of irradiated cells are determined by cell cycle-dependent nuclear functions. In contrast to p53, p21waf1 was equally distributed among the nuclei of multinucleated cells regardless of the stage of the cell cycle, indicating that the observed phenomenon is specific for p53. PMID- 9393738 TI - MMAC1/PTEN mutations in primary tumor specimens and tumor cell lines. AB - A candidate tumor suppressor gene, MMAC1/PTEN, located in human chromosome band 10q23, was recently identified based on sequence alterations observed in several glioma, breast, prostate, and kidney tumor specimens or cell lines. To further investigate the mutational profile of this gene in human cancers, we examined a large set of human tumor specimens and cancer cell lines of many types for 10q23 allelic losses and MMAC1 sequence alterations. Loss of heterozygosity (LOH) at the MMAC1 locus was observed in approximately one-half of the samples examined, consistent with the high frequency of 10q allelic loss reported for many cancers. Of 124 tumor specimens exhibiting LOH that have been screened for MMAC1 alterations to date, we have detected variants in 13 (approximately 10%) of these primary tumors; the highest frequency of variants was found in glioblastoma specimens (approximately 23%). Novel alterations identified in this gene include a missense variant in a melanoma sample and a splicing variant and a nonsense mutation in pediatric glioblastomas. Of 76 tumor cell lines prescreened for probable LOH, microsequence alterations of MMAC1 were detected in 12 (approximately 16%) of the lines, including those derived from astrocytoma, leukemia, and melanoma tumors, as well as bladder, breast, lung, prostate, submaxillary gland, and testis carcinomas. In addition, in this set of tumor cell lines, we detected 11 (approximately 14%) homozygous deletions that eliminated coding portions of MMAC1, a class of abnormality not detected by our methods in primary tumors. These data support the occurrence of inactivating MMAC1 alterations in multiple human cancer types. In addition, we report the discovery of a putative pseudogene of MMAC1 localized on chromosome 9. PMID- 9393740 TI - Evidence that the multidrug resistance protein (MRP) functions as a co transporter of glutathione and natural product toxins. AB - The MRP (multidrug resistance protein) gene, a member of the ubiquitous superfamily of ATP-binding cassette transporters, is associated with the multidrug resistance of mammalian cells to natural product anticancer agents. We have previously shown that abrogation of MRP expression by gene targeting leads to hypersensitivity to several drugs. In two independently produced MRP double knockout clones, the baseline export of glutathione (GSH) was one-half that of wild-type embryonic stem (ES) cells. The export of GSH from wild-type ES cells, but not from the MRP double knockout clones, increased in the presence of etoposide (VP-16) and sodium arsenite, accompanied by equivalent decreases in intracellular levels of GSH. In the two MRP double knockout clones, the intracellular steady-state concentration of etoposide was twofold greater than that in wild-type cells. Depletion of intracellular GSH by D,L-buthionine sulfoximine increased the intracellular accumulation of radiolabeled etoposide in parental ES cells up to the level present in the two MRP knockout clones but did not change etoposide levels in the MRP knockout clones. These observations provide evidence that: (a) MRP exports GSH physiologically, presumably in association with an endogenous compound(s); (b) baseline MRP expression protects cells from the toxic effects of xenobiotics by effluxing the xenobiotics and GSH from the intracellular compartment into the extracellular medium by a co transport mechanism; and (c) disruption of the gene encoding MRP abrogates the cotransport of xenobiotics and GSH. PMID- 9393739 TI - Expression of PACE4 in chemically induced carcinomas is associated with spindle cell tumor conversion and increased invasive ability. AB - Gene expression changes associated with the conversion of squamous cell carcinoma (SCC) to a more advanced malignant spindle cell carcinoma (SPCC) were determined by differential display. Using an animal model of human SCC progression, we provide evidence of increased PACE4 expression in SPCC cell lines and primary tumors induced by chemical carcinogenesis protocols, thus implicating this proprotein convertase in the process of tumor progression. Exogenous overexpression of PACE4 cDNA in mouse SCC cells of low invasive ability resulted in enhanced tumor cell invasiveness that was absent in parental or mock transfected SCC cells. In addition, the PACE4-transfected cells acquired the ability to process prostromelysin 3 into its active enzyme form. Taken together, these results show that up-regulation of PACE4 expression is associated with SCC conversion to SPCC and suggests that activation of essential PACE4 substrates, such as the metalloproteinase stromelysin 3, is required for tumor cell invasion. PMID- 9393741 TI - Disruption of the murine MRP (multidrug resistance protein) gene leads to increased sensitivity to etoposide (VP-16) and increased levels of glutathione. AB - The mrp (multidrug resistance protein) gene has been associated with the multidrug resistance of cancer cells in vitro and in vivo. To gain information on its physiological role, embryonic stem cells were used to generate mice homozygous for a disruption of the mrp gene, resulting in complete abrogation of mrp expression. No physiological abnormalities were observed, at least up to 4 months of age. Viability, fertility, and a range of histological, hematological, and serum-chemical parameters were similar in mrp(+/+) and mrp(-/-) mice. mrp(-/ ) mice displayed an increased sensitivity to etoposide phosphate (2-fold) accompanied by greater bone marrow toxicity, whereas the acute toxicity of sodium arsenite was equivalent in mrp(+/+) and mrp(-/-) mice. Tissue levels of glutathione (GSH) were elevated in breast, lung, heart, kidney, muscle, colon, testes, bone marrow cells, blood mononuclear leukocytes, and blood erythrocytes of mrp(-/-) mice and were unchanged in organs known to express little if any mrp, such as the liver and small intestine. The increase in GSH was not due to an increase in the activity of gamma-glutamylcysteine synthetase, the rate-limiting enzyme for GSH synthesis. The findings demonstrate that mrp is dispensable for development and growth but exerts a role in drug detoxification and GSH metabolism. PMID- 9393742 TI - NADH dehydrogenase deficiency in an apoptosis-resistant mutant isolated from a human HL-60 leukemia cell line. AB - An apoptosis-resistant mutant (VC-33) was selected from HL-60 by alternating exposure to camptothecin and etoposide. VC-33 cells demonstrated resistance to apoptosis as induced not only by camptothecin and etoposide but by a variety of other agents as well, including 1-beta-D-arabinofuranosylcytosine, hydroxyurea, calcium ionophore (A23187), cycloheximide, and UV irradiation. In an effort to identify the mechanism of such apoptosis resistance, a mRNA differential display analysis was used. Among a total of 12 bands with reduced expression in VC-33 cells, 1 cDNA clone was isolated that was hybridized to the wild-type transcript but not to the VC-33 transcript on Northern blotting. Partial sequence of this gene revealed 98% homology to mitochondrial NADH dehydrogenase subunit 5. When cell growth and intracellular ATP levels under glucose starvation were measured, VC-33 cells were found to be more sensitive than wild-type cells. Thus, NADH dehydrogenase deficiency may contribute, at least in part, to the mechanism of resistance to apoptosis in VC-33 cells. PMID- 9393743 TI - Liposomal doxorubicin circumvents PSC 833-free drug interactions, resulting in effective therapy of multidrug-resistant solid tumors. AB - Conventional methods that are used to overcome multidrug resistance (MDR) often involve the coadministration of chemosensitizers and anticancer drugs. The cyclosporin analogue SDZ PSC 833 [(3'-keto-Bmt1)-(Val2)-cyclosporin] (PSC 833) has been shown to possess powerful chemosensitization properties in vitro, in addition to being intrinsically nontoxic. However, coadministration of PSC 833 with anticancer drugs, such as daunorubicin, doxorubicin (DOX), and Taxol, have resulted in the exacerbation of anticancer drug toxicity, which is due to altered anticancer drug pharmacokinetics. Here, we hypothesized that optimization of the anticancer drug delivery, using liposomal carriers, may, by avoiding these adverse interactions, offer a significant advantage over nonencapsulated drugs. Toxicity studies were conducted in normal BDF1 mice, with i.v. DOX (free or liposome encapsulated) administration and p.o. PSC 833 in single and multiple dosage regimens over a 15-day study period. p.o. administration of PSC 833, at a dose of 100 mg/kg, reduced the maximum tolerated dose (MTD) of i.v administered free drug by 2.5-3-fold, in single- and multiple-dose regimens. In contrast, PSC 833 administration resulted in only a 20% reduction of the MTD for DOX encapsulated in 100-nm 1,2 distearoyl-sn-glycero-3-phosphocholine/cholesterol liposomes (55:45 molar lipid ratio) in a single-dose regimen and had no effect on the liposomal DOX MTD for the day 1, 5, and 9 treatment schedule. Modest modulation of P-glycoprotein-mediated MDR was observed in the murine P388/ADR solid tumor model when PSC 833 was administered with free DOX at the MTD. In contrast, liposomal DOX combined with PSC 833 resulted in tumor growth inhibition that was comparable to that observed for drug-sensitive P388/WT tumors. This efficacy of P388/ADR tumors treatment was dependent on PSC 833 because treatment with liposomal DOX alone provided significantly less antitumor activity. Pharmacokinetic and tissue distribution data demonstrated that DOX encapsulated in 1,2 distearoyl-sn-glycero-3-phosphocholine/cholesterol liposomes exhibited comparable plasma elimination and tissue distribution properties in the presence and absence of PSC 833, whereas free DOX displayed reduced plasma elimination rates and altered tissue distribution in the presence of PSC 833. These results provide evidence that PSC 833 can induce P-glycoprotein modulation and chemosensitize MDR tumors in the absence of altered DOX pharmacokinetics when liposomal carriers are used. This suggests that the improved tumor selectivity of anticancer drugs that are administered in liposomal formulations may avoid the complications that are associated with free drug-MDR-reversing agent combinations and enhance the therapy of multidrug-resistant tumors. PMID- 9393744 TI - PTEN/MMAC1 mutations and EGFR amplification in glioblastomas. AB - Loss of heterozygosity (LOH) from chromosome 10 is a hallmark of glioblastoma, the most malignant (grade IV) form of glioma. A candidate tumor suppressor gene, PTEN/MMAC1, that may be targeted for deletion in association with chromosome 10 LOH has recently been identified. Here we have investigated 63 glioblastomas for PTEN/MMAC1 alterations and identified DNA sequence changes that would affect the encoded protein in 17 (27%) tumors. Microsatellite analyses of normal-tumor DNA pairs were performed on 14 of these cases and revealed LOH at locations flanking and/or near PTEN/MMAC1 in all but 1 instance, suggesting that deletion of the remaining wild-type allele had occurred in the large majority of tumors with PTEN/MMAC1 mutations. Competitive PCR assays were developed to address the possible occurrence of PTEN/MMAC1 homozygous deletions in glioblastomas, and this analysis identified three samples having loss of both PTEN/MMAC1 alleles. EGFR amplification was determined to occur at similar frequencies among cases with or without PTEN/MMAC1 homozygous deletions or mutations, suggesting that a growth promoting effect resulting from amplification-associated increases in epidermal growth factor receptor signaling is not necessarily dependent on the inactivation of PTEN/MMAC1. PMID- 9393745 TI - The use of exfoliative cell samples to map clonal genetic alterations in the oral epithelium of high-risk patients. AB - Although it is widely accepted that clonal genetic alterations are an essential component of tumor progression, little is known of the distribution of such changes in high-risk lesions or how such clones are altered over time. We explored the feasibility of using exfoliative cells collected by scraping the mucosal surface to detect allelic loss in oral lesions of 22 patients (14 squamous cell carcinomas, 2 carcinomas in situ, and 6 dysplasias). The data show that the patterns of allelic loss observed in these samples closely represent those observed in biopsies of the same region. Furthermore, early indications are that this approach can be used to detect recurrent outgrowth of clones of altered cells in patients after therapy. PMID- 9393746 TI - Human tumor blood flow is enhanced by nicotinamide and carbogen breathing. AB - Perfusion insufficiency and the resultant hypoxia are recognized as important mechanisms of resistance to anticancer therapy. Modification of the tumor microenvironment to increase perfusion and oxygenation of tumors may improve on the efficacy of these treatments. Using laser Doppler probes to measure microregional RBC flux, this study examines the influence of nicotinamide and carbogen on human tumor perfusion. Ten patients with advanced cancers were studied. Nicotinamide (80 mg/kg) was given p.o., and 60 min later, up to six probes were inserted into the tumor. Readings were taken for 1 h, followed by 10 min of carbogen breathing and 10 additional min of breathing room air. Results were compared with those from a similar group of eight control patients who were not given nicotinamide, but who breathed carbogen. In 44 microregions analyzed, 33 (73%) showed perfusion fluctuations of 50% or more, and 20 (44%) by 100% or more. This compared with the control group in whom 62% and 27% of microregions varied by 50% or more and 100% or more, respectively. Perfusion increases outweighed decreases by 30% with nicotinamide and 20% in the controls. On breathing carbogen, patients pretreated with nicotinamide showed an increase in tumor perfusion of 17% at 5 min and 22% at 10 min, compared with only 0% and 1% in the control group. Pretreatment with nicotinamide made little difference to the random blood flow fluctuations seen in controls. However, when carbogen was introduced, tumor perfusion increased compared with the control group. This may have important therapeutic implications by improving response to treatment and allowing better delivery of systemically administered agents. PMID- 9393747 TI - Overexpression of manganese superoxide dismutase selectively modulates the activity of Jun-associated transcription factors in fibrosarcoma cells. AB - Manganese superoxide dismutase (MnSOD) is reduced in a variety of tumor cells and has been proposed to be a new type of tumor suppressor gene. The mechanism(s) by which MnSOD suppresses cancer development is currently unknown. However, expression of this antioxidant might play a significant role in maintaining cellular redox status. The relationship between MnSOD expression and modulation of DNA-binding activity and transcriptional activation of redox-sensitive oncoproteins and tumor suppressor proteins was studied in a murine fibrosarcoma cell line (FSa-II). Electrophoretic mobility shift assay and transcriptional activation studies revealed an inverse correlation between MnSOD expression and activity of c-jun-associated transcription factors, activator protein 1 and cyclic AMP-responsive element binding protein. Furthermore, expression of an activator protein 1 target gene, bcl-xL, was decreased in MnSOD-transfected cell lines. The results suggest that overexpression of MnSOD may exert its tumor suppressor activity, in part, by modulation of specific oncogenes. PMID- 9393748 TI - Galectin-3: a novel antiapoptotic molecule with a functional BH1 (NWGR) domain of Bcl-2 family. AB - Galectin-3, a beta-galactoside-binding protein, has been shown to be involved in tumor progression and metastasis. Here, we demonstrate that expression of galectin-3 in human breast carcinoma BT549 cells inhibits cis diamminedichloroplatinum (cisplatin)-induced poly(ADP-ribose) polymerase degradation and apoptosis, without altering Bcl-2, Bcl-X(L), or Bax expressions. Galectin-3 contains the NWGR amino acid sequence highly conserved in the BH1 domain of the bcl-2 gene family, and a substitution of glycine to alanine in this motif abrogated its antiapoptotic activity. Our findings demonstrate that galectin-3 inhibits apoptosis through a cysteine protease pathway and highlight the functional significance of the NWGR motif in apoptosis resistance of a non Bcl-2 protein. PMID- 9393749 TI - Human angiostatin inhibits murine hemangioendothelioma tumor growth in vivo. AB - Angiostatin inhibits angiogenesis and metastatic tumor growth; however, its usefulness in treating primary nonmetastasizing tumors is less well understood. We now report the effectiveness of human angiostatin administration in a mouse hemangioendothelioma model. Human angiostatin was administered to mice with s.c. hemangioendothelioma and associated disseminated intravascular coagulopathy (Kasabach-Merritt syndrome). Angiostatin significantly reduced tumor volume in comparison to nontreated controls, increased survival, and prevented the profound thrombocytopenia and anemia of Kasabach-Merritt syndrome. Apoptosis of tumor cells was induced by angiostatin, but tumor cell proliferation was not inhibited. These data suggest angiostatin as a novel treatment for nonmetastasizing vascular tumors and for Kasabach-Merritt syndrome. PMID- 9393750 TI - Vascular endothelial growth factor is a predictor of relapse and stage progression in superficial bladder cancer. AB - Tumor development is angiogenesis dependent, and vascular endothelial growth factor (VEGF) is a key growth factor in this process. We demonstrate that high expression of VEGF mRNA in 55 superficial bladder cancers was associated with earlier recurrence (P = 0.001; hazard ratio, 3.09) and progression to a more invasive phenotype (P = 0.02; hazard ratio, 5.33). VEGF mRNA expression correlated with protein levels in superficial tumors (r = 0.59, P = 0.003) and normal bladder (r = 0.65, P < 0.05), although the ratio of VEGF protein to mRNA was elevated in tumors compared to normal bladder (P = 0.004), suggesting posttranscriptional regulation. In this study, VEGF is implicated as a major downstream mediator of the effects of the p53 tumor suppressor gene by the association between high p53 protein (determined immunochemically) and high VEGF protein and mRNA expression (P < 0.02), although in cases without high p53 protein expression, high VEGF mRNA also predicts a poor prognosis. The relationship between VEGF and early tumor recurrence suggests that seeding via angiogenesis may be a major mechanism in the pathogenesis of recurrence. These studies indicate that VEGF can predict the behavior of superficial bladder tumors and is a therapeutic target for intravesical therapy. PMID- 9393751 TI - Murine susceptibility to radiation-induced pulmonary fibrosis is influenced by a genetic factor implicated in susceptibility to bleomycin-induced pulmonary fibrosis. AB - From evidence of interpatient variability in normal tissue sensitivity to radiotherapy and from radiation studies using inbred mouse strains, it is hypothesized that individual variation in susceptibility to radiation-induced pulmonary fibrosis is genetically controlled. A genetic model has been developed from the fibrosis-prone C57BL/6J and the fibrosis-resistant C3Hf/Kam mouse strains. Inheritance of the fibrotic phenotype was characterized in F1 and F2 (F1 intercross) generations derived from the parental strains. Genetic mapping was used to determine whether the quantitative trait loci (QTL), which influence susceptibility to bleomycin-induced lung fibrosis in these progenitor strains, could be implicated in susceptibility to radiation-induced lung fibrosis. Mice were treated with 14 or 16 Gy (60Co) to the whole thorax. The doses were selected to investigate the response at the LD50 and LD100 of C3Hf/Kam mice. The animals were sacrificed 33 weeks after treatment or when moribund. The percentage of lung with fibrosis for each mouse was quantified with image analysis of a histological section of the lung. For both the 14- and 16-Gy data sets, heritability was estimated at 38 +/- 11%, and the number of genetic factors influencing susceptibility to pulmonary fibrosis was estimated to be one or two. Two hundred fifty-five F2 intercross mice were genotyped with markers at the bleomycin loci on chromosomes 11 and 17 (chromosome 17 marker is at the major histocompatibility complex). Genetic linkage was established for the marker on chromosome 17 (P = 3.0 x 10(-6)), which accounts for 6.6% of the F2 phenotypic variance but not for the markers surrounding the QTL on chromosome 11 (P = 0.37). The inheritance data suggested that susceptibility to radiation-induced pulmonary fibrosis is a heritable trait controlled by two genetic loci, and through genomic mapping, a QTL on chromosome 17 was identified as one of the loci. PMID- 9393752 TI - Transient induction of the MRP/GS-X pump and gamma-glutamylcysteine synthetase by 1-(4-amino-2-methyl-5-pyrimidinyl)methyl-3-(2-chloroethyl)-3- nitrosourea in human glioma cells. AB - Treatment of human glioma A172 cells with 1-(4-amino-2-methyl-5 pyrimidinyl)methyl-3-(2-chloroethyl)-3-nitrosourea (ACNU), an alkylating antitumor agent the primary target of which has been thought to be DNA, resulted in elevated expression of mRNA for multidrug resistance-associated protein (MRP) within the first 2 h and then a decrease in expression 24 h after the treatment. Western blot analyses revealed that levels of MRP in these ACNU-treated cells paralleled mRNA levels. Membrane vesicles prepared from ACNU-treated cells also displayed elevated transport activities for leukotriene C4, a known substrate for MRP. Gamma-glutamylcysteine synthetase (gamma-GCS) mRNA expression was coinduced with MRP by ACNU. Because gamma-GCS is the rate-limiting enzyme involved in the de novo biosynthesis of glutathione, increases in glutathione were also transiently induced by ACNU. These results demonstrate for the first time that the expression of functional MRP and gamma-GCS can be transiently coinduced by ACNU. Multiple short exposures (1 h) of ACNU following a long duration (1 week) of drug-free conditions resulted in the development of an ACNU-resistant population (designated A172R) that overexpressed MRP/gamma-GCS mRNA and had elevated transport activities for leukotriene C4. A172R exhibited cross resistance to the antitumor drug doxorubicin and heavy metal sodium arsenate but not to cisplatin. Our results also demonstrate that intermittent treatments of human glioma cells with ACNU can lead to the development of MRP-related multidrug resistance. These results, taken together, reveal a possible new mechanism of the development of drug resistance for the antitumor nitrosoureas. PMID- 9393753 TI - The protein kinase C activators phorbol esters and phosphatidylserine inhibit neutral sphingomyelinase activation, ceramide generation, and apoptosis triggered by daunorubicin. AB - To address the role of protein kinase C (PKC) in the regulation of ceramide production, we evaluated the impact of the PKC activators 12-O tetradecanoylphorbol-13-acetate and phosphatidylserine on the apoptotic signaling pathway triggered by the chemotherapeutic drug daunorubicin. Treatment of U937 and HL-60 cells with 0.5-1 microM daunorubicin induced a greater than 30% activation of neutral sphingomyelinase activity within 4-10 min with concomitant sphingomyelin hydrolysis and ceramide generation. Activation of PKC by 12-O tetradecanoylphorbol-13-acetate and phosphatidylserine inhibited daunorubicin induced neutral sphingomyelinase activation, sphingomyelin hydrolysis, ceramide generation, and apoptosis. The apoptotic response could be restored by the addition of 25 microM cell-permeant C6-ceramide. In conclusion, PKC emerges as a potentially critical negative regulator of the anthracycline-activated sphingomyelin-ceramide apoptotic pathway. PMID- 9393754 TI - Intravenous administration of irinotecan elevates the blood beta-glucuronidase activity in rats. AB - 7-Ethyl-10-hydroxycamptothecin (SN-38) is the active metabolite of an anticancer drug, irinotecan (CPT-11). Severe late diarrhea is the dose-limiting toxic effect of CPT-11. This diarrhea has been examined regarding biliary excretion and deconjugation of SN-38 glucuronide by the enzyme beta-glucuronidase (beta-GL) in intestinal microflora. Prompted by the enzymological and structural similarity of CPT-11 to organophosphorus and carbamate insecticides, we studied the effect of CPT-11 on blood beta-GL activity in rats. The i.v. injection of CPT-11 in rats significantly elevated their plasma beta-GL activity (with phenolphthalein glucuronide as a substrate) at doses of 10 and 40 mg/kg, with peak activity observed 2-3 h after administration. SN-38 lactone and carboxylate had no effect on the plasma beta-GL level. The enhancement of the activity was also observed in serum using SN-38 glucuronide as a substrate. The serum beta-GL levels showed a close correlation between these substrates. The enhancement of plasma (serum) beta-GL activity is suggested to be a result of the release of beta-GL from liver microsomes. Serum and microsomal carboxylesterase were not significantly affected by CPT-11 administration. PMID- 9393755 TI - Improved treatment of medullary thyroid cancer in a nude mouse model by combined radioimmunochemotherapy: doxorubicin potentiates the therapeutic efficacy of radiolabeled antibodies in a radioresistant tumor type. AB - Whereas in advanced metastatic medullary thyroid cancer (MTC), a variety of chemotherapeutic regimens have achieved only limited success clinically, more recently, radioimmunotherapy (RIT) with 131I-labeled anti-carcinoembryonic antigen (CEA) monoclonal antibodies (MAbs) has shown promising results. The aims of this study were to compare, in an animal model, the therapeutic efficacy of RIT to clinically used "standard" chemotherapeutic regimens and to evaluate whether combination strategies of both modalities may be feasible and may help to improve therapeutic results in this rather radioresistant tumor type. Nude mice, bearing s.c. xenografts of the human MTC cell line, TT, were treated either with the 131I-labeled anti-CEA MAb, F023C5 IgG, or were administered chemotherapeutic regimens that had shown promising results in patients with metastatic MTC (doxorubicin and cisplatinum monotherapy, combinations of both agents, and a 5 fluorouracil/dacarbazine/streptozotocin scheme). Control groups were left untreated or were injected with an irrelevant radiolabeled antibody at equitoxic dose levels. The maximum tolerated dose (MTD) of each agent was determined. Combinations of chemotherapy and RIT were evaluated as well. Toxicity and tumor growth were monitored at weekly intervals. From the chemotherapeutic agents and schemes tested, doxorubicin monotherapy was the most effective; combination therapies did not result in an increased antitumor efficacy, but they did result in more severe toxicity. At equitoxic doses, no significant difference was found between the therapeutic efficacy of doxorubicin and that of RIT. Myelotoxicity was dose limiting with radiolabeled MAbs (MTD, 600 microCi), as well as with chemotherapeutic regimens containing alkylating agents (cisplatinum, dacarbazine, or streptozotocin). At its MTD (200 microg), doxorubicin caused only mild myelotoxicity, and despite signs of cardiac toxicity, gastrointestinal side effects were dose limiting. Accordingly, bone marrow transplantation (BMT) enabled dose intensification with RIT (MTD with BMT, 1100 microCi), which led to further increased antitumor efficacy, whereas BMT was unable to increase the MTD of doxorubicin. Due to the complementarity of toxic side effects but an anticipated synergism of antitumor efficacy, combinations of RIT with doxorubicin were tested. Administrations of 500 microCi of 131I-labeled anti-CEA and, 48 h later, 200 microg of doxorubicin (i.e., 83 and 100% of the respective single agent MTDs), were the highest doses that did not result in an increased lethality; with bone marrow support, 1000 microCi of 131I-labeled anti-CEA could be combined with 200 microg of doxorubicin (i.e., 90 and 100% of the individual MTDs). Therapeutic results of this combined radioimmunochemotherapy were superior to equitoxic monotherapy with either agent, and indication for synergistic antitumor effects is given. At its respective MTD, radioimmunochemotherapy led to a 36% cure rate if it was given without bone marrow support and to a 85% permanent cure rate if it was given with bone marrow support. The animal model, as presented in this study, seems to be useful for the preclinical testing of therapeutic agents for the systemic treatment of MTC. At equitoxic doses, RIT with radiolabeled anti-CEA antibodies seems to be equally as effective as chemotherapy with doxorubicin. Combination of RIT and doxorubicin chemotherapy seems to have synergistic therapeutic efficacy, which may be due to a radiosensitizing effect of doxorubicin. PMID- 9393756 TI - T-cell receptor repertoire in matched MART-1 peptide-stimulated peripheral blood lymphocytes and tumor-infiltrating lymphocytes. AB - Characterization of tumor-associated antigens (TAAs) recognized by CTLs makes the consideration of therapeutic strategies based on peptide stimulation of peripheral blood lymphocytes (PBLs) feasible. Several such approaches are adoptive transfer of peptide-stimulated PBLs, ex vivo peptide stimulation of dendritic cells, and direct vaccination with TAA-derived peptides. A critical component of any of these peptide-based strategies is the requirement that the patient's PBLs are able to react productively against the presented TAA. The purpose of this study, through the study of T-cell receptor (TCR) usage, was to evaluate the T-cell response in matched MART-1(27-35) peptide-stimulated PBLs and tumor-infiltrating lymphocytes (TILs). MART-1(27-35)-reactive PBL and TIL cultures were generated from three patients by in vitro stimulation with an immunodominant peptide of MART-1 (MART-1(27-35)). All cultures had a human leukocyte antigen A2-restricted, MART-1(27-35)-specific CTL response. The TCR usage of each was assessed by the DNA sequence analysis of 50 TCR beta clones obtained by rapid amplification of cDNA ends per culture. TCR analysis suggests a TCR repertoire that differed from patient to patient (8-16 subfamilies were used) and a predominant usage of a different variable beta chain (BV) by each of these MART-reactive T cells. These predominant BV rearrangements were derived from multiple clonotypes because different variable, diversity, and junctional regions were observed. However, a similar pattern of expansion was present for both PBLs and TILs; the relative usage of each prevailing BV was more marked in TILs (36, 50, and 78% of TILs versus 26, 20, and 24% of PBLs, respectively), a broader TCR repertoire was used by PBLs (P > 0.05), and similar TCR subfamily usage was noted when TIL and PBL cultures from the same patient were compared (8 of 11, 7 of 9, and 7 of 8 for patients 1, 2, and 3, respectively). Furthermore, the exact same clonotypes derived from predominant TCR subfamilies in the PBLs and TILs were present in each patient, suggesting peptide-stimulated expansion in both biological compartments. These studies suggest that there will not be a limited and predictable TCR subfamily response to a specific TAA, although reproducible patterns of PBL and TIL expansion are present from patient to patient. Additionally, identical T-cell clonotypes having the same potential for antigen driven expansion were present in a patient's PBLs and TILs. As such, our data support the conceptualization of approaches using adoptive transfer or vaccination based on TAA-derived peptide stimulation of PBLs. PMID- 9393757 TI - V-SRC induces expression of hypoxia-inducible factor 1 (HIF-1) and transcription of genes encoding vascular endothelial growth factor and enolase 1: involvement of HIF-1 in tumor progression. AB - Adaptation to hypoxia represents an important aspect of tumor progression. Hypoxia-inducible factor 1 (HIF-1) is a transcription factor that mediates essential homeostatic responses to cellular and systemic hypoxia by activating transcription of multiple genes including those encoding glycolytic enzymes and vascular endothelial growth factor (VEGF). In this report, we demonstrate that whereas C-SRC expression is not required for expression of HIF-1 or transcriptional activation of genes encoding VEGF and enolase 1 (ENO1), cells expressing the v-Src oncogene have increased expression of HIF-1, VEGF, and ENO1 under both hypoxic and nonhypoxic conditions. Expression of V-SRC was associated with increased transcription of reporter genes containing cis-acting hypoxia response elements from the VEGF and ENO1 genes, and this transcriptional activation required an intact HIF-1 binding site. When three rat hepatoma subclones that differed with respect to the level of HIF-1 expression were injected into nude mice, tumor growth correlated with HIF-1 expression, suggesting that HIF-1 may be generally involved in tumor progression. These studies link an oncogene to the induction of HIF-1 expression, thus providing a mechanism for hypoxic adaptation by tumor cells. PMID- 9393758 TI - Low frequency of p16/CDKN2A methylation in sporadic melanoma: comparative approaches for methylation analysis of primary tumors. AB - Methylation of the 5' CpG island of the p16 tumor suppressor gene represents one possible mechanism for inactivation of this cell cycle regulatory gene that is also a melanoma predisposition locus. We have investigated the potential contribution of somatic silencing of the p16 gene by DNA methylation in 30 cases of sporadic cutaneous melanoma. The methylation status of the 5' CpG island of p16 was initially determined by Southern analysis and then reevaluated (in a blinded manner) using methylation-specific PCR, methylation-sensitive single nucleotide primer extension, and bisulfite genomic sequencing. All methodologies yielded concordant results, and significant levels of methylation were observed in 3 of the 30 (10%) melanoma DNAs analyzed. Of the three tumors found to be methylated, two were also positive for LOH on 9p21 (where the p16 gene resides), implying that both p16 alleles were inactivated, one via deletion and the other via methylation-associated transcriptional silencing. The association between methylation and transcriptional silencing of p16 was also further supported by inducing p16 expression with a DNA demethylating agent (5-aza-2'-deoxycytidine) in a melanoma cell line known to harbor a methylated p16 allele. Although methylation-associated gene silencing does not represent a common mechanism for p16 inactivation in sporadic melanoma, our findings provide support that PCR based techniques, such as methylation-specific PCR and methylation-sensitive single nucleotide primer extension, can be reliably used for the accurate detection and quantitation of aberrant levels of DNA methylation in tumor specimens. PMID- 9393759 TI - Enforced expression of the Mr 33,000 Pim-1 kinase enhances factor-independent survival and inhibits apoptosis in murine myeloid cells. AB - Expression of the Mr 33,000 human Pim-1 protein is induced in hematopoietic cells by a variety of growth factors and cytokines. We have introduced the human pim-1 cDNA via retroviral transduction into interleukin (IL)-3-dependent FDC-P1 cells and examined the resulting phenotype. Compared with cells infected with a neo encoding retrovirus (FD/neo), cells infected with a pim-1-transducing virus (FD/hpim) showed longer survival or autonomous growth in suspension culture in the absence of IL-3, as well as IL-3-independent clonogenic growth in semisolid medium. The unique murine Mr 44,000 Pim-1 protein, as well as human proteins with short C- or N-terminal truncations, also was biologically active. This effect of Pim-1 expression was associated with a decrease in apoptotic cells and an increase in G0/G1-phase cells, and the increase in G0/G1-phase cells caused by enforced expression of Pim-1 was due to a decrease in apoptosis rather than to a decrease in transit of the G1-S-phase checkpoint. The Pim-1 kinase appears to function primarily as a survival factor in factor-dependent FDCP-1 cells subjected to either cytokine withdrawal or exposure to cytotoxic agents. PMID- 9393760 TI - Genomic organization and mutation analysis of Hel-N1 in lung cancers with chromosome 9p21 deletions. AB - Allelic loss of chromosome 9p21 is common in small cell lung cancer (SCLC), but inactivation of the tumor suppressor gene CDKN2a is rare, implying the existence of another target gene at 9p21. A recent deletion mapping study of chromosome 9p has also identified a site of deletion in non-small cell lung cancer (NSCLC) centered around D9S126. The Hel-N1 (human elav-like neuronal protein 1) gene encodes a neural-specific RNA binding protein that is expressed in SCLC. We have mapped this potentially important gene in lung tumorigenesis to within 100 kb of the D9S126 marker at chromosome band 9p21 by using homozygously deleted tumor cell lines and fluorescence in situ hybridization to normal metaphase spreads. Hel-N1 is, therefore, a candidate target suppressor gene in both SCLC and NSCLC. We have determined the genomic organization and intron/exon boundaries of Hel-N1 and have screened the entire coding region for mutations by sequencing 14 primary SCLCs and cell lines and 21 primary NSCLCs preselected for localized 9p21 deletion or monosomy of chromosome 9. A homozygous deletion including Hel-N1 and CDKN2a was found in a SCLC cell line, and a single-base polymorphism in exon 2 of Hel-N1 was observed in eight tumors. No somatic mutations of Hel-N1 were found in this panel of lung tumors. Hel-N1 does not appear to be a primary inactivation target of 9p21 deletion in lung cancer. PMID- 9393761 TI - Role of O6-methylguanine-DNA methyltransferase in the resistance of pancreatic tumors to DNA alkylating agents. AB - Pancreatic adenocarcinomas rarely respond to radiation or chemotherapy, indicating that a large percentage of these tumors possess complex mechanisms of resistance. The failure of alkylating agents, such as carmustine [1,3-bis(2 chloroethyl)-1-nitrosourea; BCNU], lomustine [1-(2-chloroethyl)-3-cyclohexyl-1 nitrosourea; CCNU], and streptozotocin, to yield consistent therapeutic results further suggests that one of these mechanisms may be the high expression of O6 methylguanine-DNA methyltransferase (MGMT). All 12 human pancreatic ductal adenocarcinomas assayed for MGMT activity showed unusually high levels, implying that these malignancies are efficient in repairing genotoxic O6-alkylguanine lesions induced by methylating (streptozotocin) and 2-chloroethylating (BCNU and CCNU) chemotherapeutic genotoxic agents. Immunohistochemical analysis of an additional 15 pancreatic tumors showed that high levels of MGMT protein reside in the nucleus and the cytoplasm of malignant cells. Both nuclear and cytoplasmic staining were absent in hyperplastic duct epithelium, but staining was invariably present in moderate to highly dysplastic foci and especially strong in invasive components of the tumor. With the exception of lymphocytes that were MGMT positive, acinar, ductal, and islet cells did not stain for MGMT in histologically normal pancreata. These data indicate that MGMT activity is up regulated in dysplastic epithelium, and its expression increases during tumor progression, reaching the highest levels in the invasive components of the tumor. Resistance of pancreatic tumor cells to alkylating agents was verified with four pancreatic tumor cell lines. CAPAN-2, CFPAC-1, PANC-1, and MIAPaCa-2, having MGMT levels of 1800, 987, 700, and 880 fmol/mg protein, respectively, were resistant to BCNU, but their resistance declined sharply following pretreatment with the MGMT inhibitor O6-benzylguanine (O6-BG). On the other hand, PANC-1 and MIAPaCa-2 could not be eradicated with N-methylnitrosourea (MNU) at concentrations as high as 2 mM, even when pretreated with O6-BG. These two lines were shown to be modified genetically in microsatellite sequences by MNU and are believed to have a defective mismatch repair system, which may explain their resistance to methylating agents. Failure of pancreatic tumors to respond to nitrosoureas is related to high levels of MGMT expression and in some cases to genomic instability. However, these tumors can be sensitized to chloroethylating drugs and eradicated following the elimination of MGMT activity by O6-BG or homologous MGMT inhibitors. PMID- 9393762 TI - Fibroblast growth factor receptor 2 limits and receptor 1 accelerates tumorigenicity of prostate epithelial cells. AB - Progressive loss of the differentiated phenotype and communication with stroma accompanies the transition of nonmalignant rat prostate epithelial cells to anaplastic, malignant tumors. Here we show that cell surface expression of the fibroblast growth factor receptor 2 (FGFR2) tyrosine kinase is reduced in malignant tumor cell populations (type II) and undetectable at the mRNA level in 30% of cells. This is in addition to the irreversible loss by splice switching of the FGFR2 ectodomain that abrogates response to FGF-7 and homologues from the stroma. One hundred % of type II malignant cells express FGFR1, which is normally expressed in the stroma. Expression of the FGFR1 kinase in premalignant type I tumor epithelial cells by transfection accelerated progression to the malignant phenotype. In contrast to the FGFR2 kinase fused to the ectodomain of FGFR1, the FGFR1 kinase failed initially to support a mitogenic response to FGF-2 in type I tumor cells. However, the FGFR1-transfected cells acquired a mitogenic response after extensive proliferation of the cell population. Resident FGFR2 and ectopic FGFR1 appeared to be partitioned in the type I cells, because neither full-length nor truncated isoforms of FGFR1 affected the mitogenic response of the other. Restoration of the FGFR2IIIb kinase to malignant cells expressing FGFR1 depressed tumor growth rates, restored responsiveness to stromal cells, and restored epithelial cell differentiation. These observations reveal that homologous FGFR1 and FGFR2 kinases play very different roles in cell growth and differentiation and in development and support of the malignant phenotype. PMID- 9393763 TI - Molecular evidence that most but not all carcinosarcomas of the uterus are combination tumors. AB - The pathogenesis of carcinosarcoma is still a subject of controversy. In the present study, molecular techniques were applied to determine the pathogenesis of uterine carcinosarcomas. The patterns of chromosome X inactivation were analyzed, targeting a portion of exon 1 of the human androgen receptor (HUMARA) in malignant epithelial and mesenchymal components. The presence of p53 and K-ras mutations were also analyzed. H&E-stained sections of paraffin-embedded, formalin fixed tissues were microdissected to obtain both epithelial and nonepithelial lesions from 25 carcinosarcomas, and DNAs were extracted by proteinase K digestion. Following treatment with methylation-sensitive restriction endonuclease (HhaI or HpaII), PCR amplification was performed using nested primers targeted to the HUMARA locus. Mutations in the p53 gene and K-ras gene were found in eight (32%) and six (24%) tumors, respectively. The patterns of chromosome X inactivation were different between the carcinomatous and sarcomatous components of three carcinosarcomas, indicating that these three tumors represent collision tumors. By contrast, the patterns of chromosome X inactivation, K-ras sequence, and p53 sequence were identical in both carcinomatous and sarcomatous components in 21 carcinosarcomas, indicating that these 21 tumors represent combination tumors. One case produced equivocal results that precluded determination of whether it represented a collision or combination tumor. These observations show that although most carcinosarcomas are combination tumors, some develop as collision tumors. The determination of histogenesis in individual cases of carcinosarcoma using molecular markers may be worthwhile, because the result could help predict the prognosis of individual cases and help guide clinical management. PMID- 9393764 TI - The osteoclast-associated protease cathepsin K is expressed in human breast carcinoma. AB - Human cathepsin K is a novel cysteine protease previously reported to be restricted in its expression to osteoclasts. Immunolocalization of cathepsin K in breast tumor bone metastases revealed that the invading breast cancer cells expressed this protease, albeit at a lower intensity than in osteoclasts. In situ hybridization and immunolocalization studies were subsequently conducted to demonstrate cathepsin K mRNA and protein expression in samples of primary breast carcinoma. Expression of cathepsin K mRNA was confirmed by reverse transcription PCR and Southern analysis in a number of human breast cancer cell lines and in primary human breast tumors and their metastases. As this protease is known to degrade extracellular matrix, including bone matrix proteins, it is possible that cathepsin K may contribute to the invasive potential of breast cancer cells, including those that metastasize to bone. Thus, cathepsin K may be a potential target leading to the design of novel drugs for cancer therapy. PMID- 9393765 TI - Met and hepatocyte growth factor/scatter factor expression in human gliomas. AB - Using double immunofluorescence staining and quantitative confocal laser scan microscopy, we show that the intensity of hepatocyte growth factor/scatter factor (HGF/SF) and Met staining in human primary brain tumors increases with the grade of malignancy and is prevalent in both the infiltrating tumor cells and endothelial hyperplastic areas. HGF/SF and Met also are expressed in vitro in glioblastoma multiforme cell lines as well as in normal human astrocyte (NHA) cells. Moreover, HGF/SF stimulates tyrosine phosphorylation of Met in both glioma cell lines and NHA cells, but only the glioma cell lines proliferate and become motile and invasive in response to HGF/SF, whereas the NHA cells are nonresponsive. These results implicate autocrine/paracrine Met-HGF/SF signaling in glioma tumorigenesis and suggest that HGF/SF signaling through Met is negatively regulated in NHA cells. PMID- 9393766 TI - Expression of multiple endocrine neoplasia 2B RET in neuroblastoma cells alters cell adhesion in vitro, enhances metastatic behavior in vivo, and activates Jun kinase. AB - Point mutations, deletions, and recombinations of the RET proto-oncogene are associated with several inherited human diseases of neural crest-derived cells: Hirschsprung's disease, familial medullary thyroid carcinoma, and the multiple endocrine neoplasia (MEN) syndromes, types 2A and 2B. RET expression is restricted to normal and malignant cells of neural crest origin, such as human neuroblastoma cells. To better understand the role of the activated RET oncogene in neural crest cells, we transfected two adherent human neuroblastoma tumor cell lines with oncogenic MEN2 mutant RET cDNAs. Transfectant clones from both cell lines overexpressing MEN2B RET demonstrated a marked increase in the cell fraction growing in suspension. Both control and MEN2B cells formed tumors at the site of injection in all cases. However, mice injected with MEN2B cells developed lung metastases at a much higher frequency than control mice. Only RET protein derived from MEN2A transfectant cells had increased autokinase activity, whereas MEN2B transfectant cells demonstrated selective activation of the mitogen activated protein kinase, Jun kinase-1 (Jnk1). These results indicate a biochemical signaling pathway that may link oncogenic RET with the metastatic process. PMID- 9393767 TI - Methyl-p-hydroxyphenyllactate-esterase activity in breast cancer: a potentially new prognostic factor in short-term follow-up. AB - We assayed methyl-p-hydroxyphenyllactate esterase (MeHPLAase) activity in 48 cases of primary breast cancer. MeHPLAase activity did not show significant correlation with estrogen receptor and progesterone receptor levels. No significant relationship was found between enzymatic activity and tumor diameter, lymph node status, mitotic activity, degree of nuclear differentiation, and proportion of the S-phase fraction. During the follow-up period (median, 18.8 months; range, 6-69 months), recurrences were observed in 18 of 48 (37%) cases. The Weibull survival regression model using the enzymatic activity as a continuous covariate showed that levels of enzymatic activity were directly associated with the risk of recurrence (P = 0.02). Assuming the mean value of enzymatic activity as the cutoff value, we found a statistically significant relationship between high MeHPLAase activity and shorter recurrence-free survival. On multivariate analysis, MeHPLAase activity proved to be an independent factor for predicting a short period of recurrence-free survival. PMID- 9393768 TI - Epidermal growth factor regulates protein kinase A activity in murine fibrosarcoma cells: differences between metastatic and nonmetastatic tumor cell variants. AB - The interplay between cyclic AMP (cAMP)-dependent protein kinase A (PKA)- and p21ras-mediated signaling pathways is expected to determine further loss, maintenance, or modulation of differentiation and proliferation of a particular cell. Therefore, the relationship and nature of the cross-talk between these two major signaling systems are of utmost importance to the understanding of these processes in both normal and neoplastic cells. In view of their paramount physiological importance, one would expect the existence of a well-controlled bidirectional interaction between these pathways, which would be more appropriate and in agreement with basic principles of cellular homeostasis. However, based on the discovery that activated PKA may inhibit ras-mediated translocation of c-Raf 1 to the plasma membrane, it is generally accepted that the cross-talk between cAMP/PKA and p21ras-mediated signal transduction pathways is unilateral, i.e., that the activation of PKA regulates growth factor receptor protein tyrosine kinase-mediated signaling. To challenge the validity of a unilateral approach, we decided to test the possible existence of cross-talk of a bidirectional nature between the aforementioned signaling pathways at different stages of malignant differentiation. For that purpose, we investigated the nature of the cross-talk existing between a known receptor protein tyrosine kinase-epidermal growth factor receptor (EGFR) and PKA in highly metastatic and nonmetastatic cloned variants of a murine fibrosarcoma (T-10). Our study revealed the existence of principal differences in PKA activity between metastatic and nonmetastatic cloned fibrosarcoma variants that may be due to the differential expression and membrane translocation of the p21(Ki-ras) small mass G-protein. Most importantly, our experiments have demonstrated the existence of a novel character of interactions between EGFR and PKA, because the ligation of the EGFR by epidermal growth factor in the metastatic variant induced a high activity of PKA. These findings are of prime importance, because they reveal the existence of a new relationship between two major signal transduction pathways in mammalian cells, i.e., the existence of a bilateral interaction between the ras- and cAMP/PKA-mediated signal transduction pathways. Furthermore, the fact that two tumor cell variants originating in the same tumor and differing in their metastatic capacity differ as well in the nature of the cross-talk between major signal generation systems imposes new challenges for the future use of biological response modulators to cure cancer and restrict metastatic spread. PMID- 9393769 TI - Fibronectin secretion from human peritoneal tissue induces Mr 92,000 type IV collagenase expression and invasion in ovarian cancer cell lines. AB - Our previous study showed that human peritoneal conditioned medium (CM) increased the matrix metalloproteinase-9 (MMP-9) secretion and invasiveness of ovarian cancer cells (NOM1). In an effort to identify this MMP-9-stimulating factor, we examined the effects of extracellular matrix components, such as type IV collagen, laminin, and fibronectin, on ovarian cancer cells. We found that fibronectin increased the MMP-9 activity of NOM1 cell CM in a concentration dependent manner and that the peritoneal CM contained high level of fibronectin. An increase of MMP-9 activity in NOM1 cell CM by the peritoneal CM was almost completely blocked by 20 microg/ml of anti-integrin alpha5/FnR antibody and RGD polypeptides. Furthermore, after immunoprecipitation by antifibronectin antibody supernatant of the peritoneal CM did not increase MMP-9 activity in NOM1 cells. Fibronectin and the peritoneal CM also increased MMP-9 activity and expression in NOM1 cell lysate, and these effects were blocked by anti-integrin alpha5/FnR antibody. Invasiveness of NOM1 cells was enhanced by fibronectin and the peritoneal CM in a concentration-dependent manner, and anti-integrin alpha5/FnR antibody blocked these effects. These results suggested that fibronectin secreted from peritoneum increased MMP-9 activity and expression, and, in turn, invasiveness of ovarian cancer cells. PMID- 9393770 TI - Vascular endothelial growth factor up-regulates its receptor fms-like tyrosine kinase 1 (FLT-1) and a soluble variant of FLT-1 in human vascular endothelial cells. AB - The growth of solid tumors and the formation of metastases are dependent on neoangiogenesis. One of the most important factors in inducing the formation of new blood vessels is the vascular endothelial growth factor (VEGF), which acts specifically on endothelial cells. VEGF is expressed and secreted by almost all solid tumors. The molecular mechanisms leading to enhanced production of this angiogenic mitogen are manyfold and have been elucidated to some degree. Two VEGF receptors, fms-like tyrosine kinase 1 (FLT-1) and KDR, have been identified almost specifically on human endothelial cells. They are expressed preferentially in the proliferating endothelium of vessels lining and/or penetrating solid tumors, whereas they are almost undetectable by convenient methods in vessels of healthy tissue. However, the underlying mechanisms are not understood. We could show that media conditioned by various cancer cell lines grown under hypoxic conditions were able to up-regulate expression of FLT-1 mRNA and protein but not of KDR mRNA. Furthermore, up-regulation of a shorter mRNA species was observed that most probably codes for the soluble variant of FLT-1. These effects were completely inhibited by VEGF-neutralizing extracellular VEGF receptor domains. The effect could be mimicked by adding recombinant VEGF instead of conditioned cancer cell medium to the endothelial cell cultures. Both mutant VEGF, which activates only KDR, and placenta growth factor, which activates only FLT-1, were able to enhance FLT-1 expression. VEGF-stimulated FLT-1 mRNA expression was inhibited by actinomycin D. These data suggest that VEGF itself is the main factor secreted by tumor cells that is able to enhance the expression of its receptor FLT-1 and of a soluble variant of FLT-1 in endothelial cells. PMID- 9393771 TI - Posttranscriptional regulation of protein expression in human epithelial carcinoma cells by adenine-uridine-rich elements in the 3'-untranslated region of tumor necrosis factor-alpha messenger RNA. AB - Eukaryotic mRNAs contain 3'-untranslated regions (UTR) that are involved in posttranscriptional control of gene expression. AU-rich octanucleotide repeats, UUAUUUAU, present in the 3'-UTR of mature lymphokine and other cytokine transcripts, have been implicated in the regulation of mRNA stability and translational efficiency. For example, previous evidence suggests that the AU rich element (ARE) present in the 3'-UTR of murine tumor necrosis factor-alpha (TNF-alpha) can affect the posttranscriptional regulation of murine TNF-alpha gene expression in hematopoietic cells. Although cytokines are produced in epithelial cells, little is known about the regulation of TNF-alpha and other cytokine gene expression by 3'-UTR elements in human malignant epithelial cells. To better understand the function of the 3'-UTR of the human TNF-alpha gene in the regulation of TNF-alpha protein production in human epithelial cancer cells, a series of luciferase reporter constructs with portions of the 3'-UTR of human TNF-alpha was transfected into human breast carcinoma cell lines ZR-75-1 and ZR 75-1R (which overexpresses TNF-alpha). The 3'-UTR of TNF-alpha markedly suppressed luciferase activity in both cell lines, and the suppression of activity was reversed by deletion of the AU-rich sequences. This suppression was quantitative, with six repeats causing more inhibition than two repeats. Increased levels of luciferase activity were observed 3 h after TNF-alpha stimulation in ZR-75-1 cells transfected by constructs containing AU-rich repeats. In addition, cytoplasmic extracts from both cell lines were assayed for factors that bind to the 3'-UTR of human TNF-alpha mRNA. RNA-protein binding activities were found in both cell lines. Competition studies showed that these proteins specifically bound to AU-rich repeats present in the 3'-UTR of TNF alpha. No binding activity was observed when the AU-rich repeats were deleted. TNF-alpha exposure markedly increased activity of several RNA-binding proteins, especially a novel Mr 50,000-55,000 RNA-binding protein. The binding activity in untreated ZR-75-1R was higher than that in untreated ZR-75-1 cells, suggesting that the level of RNA-protein binding correlates with the expression level of TNF alpha in human epithelial cancer cells and that the RNA-binding proteins may control expression of TNF-alpha in ZR-75-1 cells. We conclude that the AU-rich repeats in the 3'-UTR of human TNF-alpha mRNA may regulate gene expression in human epithelial cancer cells by binding to AU sequence-specific proteins, including a previously undescribed Mr 50,000-55,000 protein not observed in hematopoietic cells. PMID- 9393772 TI - Correspondence re: J. S. DeFrank et al., p53-null cells are more sensitive to ultraviolet light only in the presence of caffeine. Cancer Res., 56: 5365-5368, 1996. PMID- 9393774 TI - Role of gamma interferon in natural clearance of Bordetella pertussis infection. AB - Using a mouse model of Bordetella pertussis infection, we have analyzed the role of gamma interferon (IFN-gamma) in bacterial clearance from the respiratory tract. Adult BALB/c mice began to clear a respiratory infection within 3 weeks postinfection, with complete resolution of infection 6 to 8 weeks postinfection. In contrast, neither adult SCID mice (which lack mature B and T lymphocytes) nor adult nude mice (which lack mature T lymphocytes) controlled B. pertussis infection, and both strains died within 3 to 5 weeks postinfection. Short-term T cell lines generated from the draining lymph nodes of the lungs of infected BALB/c mice were found to be CD4+ and produced IFN-gamma but no detectable interleukin-4. Analyses of IFN-gamma mRNA induction in the lungs of mice following B. pertussis infection showed that in both BALB/c and C57BL/6 mice, IFN gamma mRNA levels increased sharply by 1 week postinfection and then subsequently declined. Further exploration of a potential role for IFN-gamma demonstrated that infection of adult BALB/c mice depleted of IFN-gamma in vivo with anti-IFN-gamma monoclonal antibodies resulted in greater numbers of bacteria recovered from the lungs than in infected, control BALB/c mice, although IFN-gamma-depleted mice could subsequently clear the infection. Infection of mice which have a disrupted IFN-gamma gene resulted in bacterial clearance with a time course similar to those seen with IFN-gamma-depleted mice. These results indicate that IFN-gamma plays a role in controlling B. pertussis infection. PMID- 9393773 TI - Interleukin-12 is critical for induction of nitric oxide-mediated immunosuppression following vaccination of mice with attenuated Salmonella typhimurium. AB - Studies from our laboratory have shown that infection of mice with an attenuated strain of Salmonella typhimurium causes a marked suppression in the capacity of splenocytes to generate an in vitro plaque-forming cell (PFC) response to sheep erythrocytes. The suppression has been shown to be mediated by mature, adherent macrophages (Mphis) and nonadherent, precursor Mphis. Nitric oxide has been identified as the suppressor factor. The present study investigated the role of interleukin-12 (IL-12) in the generation of nitric oxide-mediated immunosuppression in this model. Salmonella inoculation resulted in marked suppression of PFC responses and high levels of nitrite production. When mice were treated with anti-IL-12 prior to inoculation, nitrite levels in splenocyte cultures were reduced by 75% and the suppression of PFC responses was prevented. The nonadherent splenocyte fraction from Salmonella-inoculated mice, which contains precursor Mphis and is weakly immunosuppressive, was treated with IL-12 in vitro. IL-12 augmented the capacity of this fraction to suppress PFC responses by normal splenocytes in a coculture system. Additionally, IL-12 induced nitrite and gamma interferon (IFN-gamma) production in a dose-dependent manner. Treatment with anti-IFN-gamma blocked nitrite production and suppression, indicating that IFN-gamma is an important intermediary in the pathway of IL-12-induced immunosuppression. These results indicate that IL-12 is critical for the induction of nitric oxide-mediated immunosuppression following S. typhimurium inoculation and, through its ability to stimulate IFN-gamma production, can induce nitric oxide-producing suppressor Mphis. PMID- 9393775 TI - A gene homologous to Saccharomyces cerevisiae SNF1 appears to be essential for the viability of Candida albicans. AB - The SNF1 gene of Saccharomyces cerevisiae (ScSNF1) is essential for the derepression of catabolic repression. We report here the isolation and characterization of an SNF1 homolog from Candida albicans (CaSNF1) which is apparently essential for the viability of this organism. The putative amino acid sequence of CaSNF1 has 68% identity with that of ScSNF1 and can restore the S. cerevisiae snf1 delta mutant's ability to utilize sucrose. Disruption of one of the CaSNF1 alleles resulted in morphological changes and decreased growth rates but did not modify the carbon source utilization pattern. Repetitive unsuccessful attempts to generate a snf1/snf1 homozygote by disruption of the second allele, using various vectors and approaches, suggest the lethal nature of this mutation. Integration into the second allele was possible only when a full-length functional SNF1 sequence was reassembled, further supporting this hypothesis and indicating that the indispensability of Snf1p prevented the isolation of snf1/snf1 mutants. The mutant bearing two disrupted SNF1 alleles and the SNF1 functional sequence maintained its ability to utilize sucrose and produced stellate colonies with extensive hyphal growth on agar media. It was demonstrated that in a mouse model, the virulences of this mutant and the wild-type strain are similar, suggesting that hyphal growth in vitro is not an indicator for higher virulence. PMID- 9393776 TI - A strategy for rational design of fully synthetic glycopeptide conjugate vaccines. AB - The present study describes a strategy to rationally design fully synthetic glycopeptide conjugate vaccines. Glycopeptide immunogens were constructed by coupling synthetic oligosaccharides comprising repeating units of synthetic 3 beta-D-ribose-(1-1)-D-ribitol-5-phosphate (sPRP) to synthetic peptides containing potent T-helper cell determinants and B-cell epitopes of the Haemophilus influenzae type b (Hib) outer membrane proteins (OMPs) P1, P2, and P6. Rabbit immunogenicity studies revealed that some of these fully synthetic glycoconjugates were capable of eliciting high titers of both anti-PRP and anti OMP immunoglobulin G antibodies. In addition, we systematically investigated the factors which could influence their immunogenicity. We observed that the magnitude of the anti-PRP antibody response markedly depended on the relative spatial orientation of sPRP and T-cell epitopes, the anti-PRP antibody response was enhanced when a multiple antigenic peptide was used as a carrier, the anti PRP antibody response was optimal for three PRP repeating units, and lipidation of peptide-PRP conjugates had a minimal effect on the magnitude of the anti-PRP antibody response. The results of this study clearly demonstrate that coupling a carbohydrate hapten to a peptide can provide T-cell help and convert it into a T cell-dependent antigen. The antisera raised against these conjugates were also found to be protective against Hib infection in the infant rat model of bacteremia. PMID- 9393777 TI - Murine model of recurrent group G streptococcal cellulitis: no evidence of protective immunity. AB - Despite the well-known tendency of cellulitis due to beta-hemolytic streptococci to recur, little is known regarding the mechanisms of human immunity to this infection. We established cellulitis in mice by using a strain of group G streptococcus (1750) originally isolated from the bloodstream of a patient with acute cellulitis. This strain, which has been studied extensively in our laboratory, expresses M protein structurally and functionally analogous to that of group A streptococci, and we have cloned and sequenced the gene encoding this protein (emmMG1). Mice injected with 5 x 10(7) CFU of strain 1750 developed nonlethal necrotic skin and soft tissue infections that healed spontaneously after 14 to 16 days. After healing, the mice were repetitively reinoculated three times with the same challenge dose of 1750. Lesion size did not decrease in severity, size, or time to healing after repetitive challenge. The maximum lesion size and tissue concentration of microorganisms increased between the first and fourth challenges. Pretreatment of 1750 cells with opsonic antisera to MG1 diminished neither the maximum lesion size nor the time course of evolution of the lesions. Thus, in the mouse model used here, there was no evidence of acquired protective immunity to experimentally induced cellulitis. PMID- 9393778 TI - Bacterial phospholipase C upregulates matrix metalloproteinase expression by cultured epithelial cells. AB - Phospholipase C (PLC) is a putative virulence factor of several pathogenic bacteria. We studied if exogenous PLC would perturb epithelial behavior in infected tissues. Gelatin and casein zymography of cell culture medium indicated that the broad-spectrum PLC of Bacillus cereus induced matrix metalloproteinase (MMP) production in epithelial cells of human skin (NHEK), human gingiva (HGE), and porcine periodontal ligament (PLE). In all three cell types, the strongest increase (ninefold) at 0.1 U/ml was seen in the MMP-9 (92-kDa gelatinase) activity, and the effect was dose dependent in the range of 0.1 to 1.0 U/ml. A relatively weaker increase (twofold) in MMP-2 (72-kDa gelatinase) was also observed in each cell type. PLC induction of MMP-3 (48-kDa stromelysin) was also seen in NHEK and HGE on gelatin and more sensitively for PLE by casein zymography (fivefold). Total gelatinolytic activity as measured by degradation of 14C labeled denatured type I collagen increased by about 18-fold (NHEK), 12-fold (HGE), and 14-fold (PLE). Northern analysis showed a clear increase in the MMP-9, and a minor increase in MMP-3 mRNA levels but no significant increase in MMP-2 mRNA levels. Further studies with PLE revealed that MMP-9 induction by PLC progressively increased with the length of cell culture time in the absence of serum. PLC induction of MMPs was polar, with MMP-9 and MMP-3 secreted primarily in the apical direction and MMP-2 secreted mainly in the basal direction. The PLC effect was blocked by neomycin, an inhibitor of the phosphoinositol signal pathway. No significant effects were observed in MMP expression with the calcium ionophore A23187 or phospholipase A2. Morphologically, PLC treatment resulted in reduced contacts between the cultured cells and loss of the cell surface microvilli. These results suggest that PLC secreted by bacterial pathogens may disrupt epithelium of infected tissue and increase the subepithelial tissue destruction through induction of MMPs. PMID- 9393779 TI - Identification of N-acetylneuraminic acid and its 9-O-acetylated derivative on the cell surface of Cryptococcus neoformans: influence on fungal phagocytosis. AB - Sialic acids from sialoglycoconjugates present at the cell surface of Cryptococcus neoformans yeast forms were analyzed by high-performance thin-layer chromatography, binding of influenza A and C virus strains, enzymatic treatment, and flow cytofluorimetry with fluorescein isothiocyanate-labeled lectins. C. neoformans yeast forms grown in a chemically defined medium contain N acetylneuraminic acid and its 9-O-acetylated derivative. A density of 3 x 10(6) residues of sialic acid per cell was found in C. neoformans. Sialic acids in cryptococcal cells are glycosidically linked to galactopyranosyl units as inferred from the increased reactivity of neuraminidase-treated yeasts with peanut agglutinin. N-Acetylneuraminic acids are alpha-2,6 and alpha-2,3 linked, as indicated by using virus strains M1/5 and M1/5 HS8, respectively, as agglutination probes. The alpha-2,6 linkage markedly predominated. These findings were essentially confirmed by the interaction of cryptococcal cells with the lectins Sambucus nigra agglutinin and Maackia amurensis agglutinin. We also investigated whether the sialyl residues present in C. neoformans are involved in the fungal interaction with a cationic solid-phase substrate and with mouse resident macrophages. Adhesion of yeast cells to poly-L-lysine was mediated, in part, by sialic acid residues, since the number of adherent cells was markedly reduced after treatment with bacterial neuraminidase. The enzymatic removal of sialic acids also made C. neoformans yeast cells more susceptible to endocytosis by macrophages. The results show that sialic acids are components of the cryptococcal cell surface that contribute to its negative charge and protect yeast forms against phagocytosis. PMID- 9393780 TI - Role of receptor binding in toxicity, immunogenicity, and adjuvanticity of Escherichia coli heat-labile enterotoxin. AB - The role of receptor binding in the toxicity, immunogenicity, and adjuvanticity of the heat-labile enterotoxin of Escherichia coli (LT) was examined by comparing native LT and LT(G33D), a B-subunit receptor binding mutant, with respect to the ability to bind to galactose and to GM1, toxicity on mouse Y-1 adrenal tumor cells, the ability to stimulate adenylate cyclase in Caco-2 cells, enterotoxicity in the patent mouse model, and oral immunogenicity and adjuvanticity. In contrast to native LT, LT(G33D) was unable to bind to the galactosyl moiety of Sepharose 4B or GM1 but did retain the lectin-like ability to bind to immobilized galactose on 6% agarose beads. LT(G33D) had no enterotoxicity in the patent mouse model but exhibited residual toxicity on mouse Y-1 adrenal tumor cells and had an ability equivalent to that of native LT to stimulate adenylate cyclase in Caco-2 cells (5,000 versus 6,900 pmol per mg of protein). In addition, LT(G33D) was unable to serve as an effective oral adjuvant for induction of immunoglobulin G or A directed against a coadministered antigen. Furthermore, LT(G33D) elicited negligible serum and mucosal antibody responses against itself. These data indicate that the toxicity, immunogenicity, and oral adjuvanticity of LT are dependent upon binding of the B subunit to ganglioside GM1. PMID- 9393781 TI - MTC28, a novel 28-kilodalton proline-rich secreted antigen specific for the Mycobacterium tuberculosis complex. AB - Proteins that are actively secreted by Mycobacterium tuberculosis serve as major targets of immune responses in the infected host. To identify and purify novel proteins in the filtrates of M. tuberculosis cultures, a bacteriophage lambda library of M. tuberculosis H37Rv DNA was immunoscreened by using an anti-culture filtrate rabbit antiserum. Of 20 positive clones isolated, 6 were analyzed and found to express the genes for two known components of the early culture filtrate, the secreted 45/47-kDa antigen complex and the KatG protein, and two novel genes. Here we report the molecular cloning and nucleotide sequence of one of the new genes encoding a culture filtrate protein of 310 amino acid (aa) residues. We called this gene mtc28. The deduced polypeptide sequence contained an NH2-terminal, highly hydrophobic 32-aa region having properties of a secretion signal peptide. The putative 278-aa mature MTC28 protein was characterized at its NH2 and COOH termini by a high content of proline and alanine residues organized in an (AP)n motif. Thus, MTC28 is a new member of a group of proline-rich antigens found in M. tuberculosis and Mycobacterium leprae. As shown by DNA hybridization experiments, the mtc28 gene was present only in species of the M. tuberculosis complex. Purified recombinant MTC28 antigen evoked strong delayed type hypersensitivity and antibody responses in guinea pigs immunized with Mycobacterium bovis BCG, but not in guinea pigs immunized with Mycobacterium avium. The strong immunological activity of MTC28 and the absence of B- and T cell epitopes cross-reactive with a common environmental mycobacterial species, such as M. avium, make this novel antigen an attractive reagent for immunodiagnosis of tuberculosis. PMID- 9393782 TI - Characterization of B-cell responses to Chlamydia trachomatis antigens in humans with trachoma. AB - The circulating B-cell responses to Chlamydia trachomatis of 60 children and 34 adults in The Gambia were characterized in a cross-sectional study of different grades of trachoma, using the enzyme-linked immunospot (ELISPOT) assay. Antibody secreting cells (ASCs) specific to chlamydial major outer membrane protein (MOMP), heat shock protein 60, and whole elementary bodies were detected in children with no evidence of ocular disease, and the immunoglobulin (IgA) response was significantly increased in those with follicular trachoma. In marked contrast, children with the most intense ocular inflammation paradoxically had an almost completely absent B-cell response of all isotypes and to all chlamydial antigens, but with normal serum IgG and IgA responses, which was even lower than in the group with no ocular inflammation. Adults with or without evidence of trachomatous scarring had equivalent numbers of circulating B cells, principally IgA, to all chlamydial antigens. Plasmablasts secreting antibodies to MOMP were present in the urine of children in the absence of urogenital infection detectable by PCR, and relative numbers were 8 to 25 times higher than in blood, suggesting site-specific homing within a common mucosal immune system. These results suggest that ELISPOT assay of ongoing B-cell responses detects suppression of chlamydia-specific IgA ASCs during the proinflammatory response to ocular chlamydial infection seen in intense trachoma, which may play a role in tissue damage leading to trachomatous scarring. PMID- 9393783 TI - Expression of the superantigen Mycoplasma arthritidis mitogen in Escherichia coli and characterization of the recombinant protein. AB - Mycoplasma arthritidis mitogen (MAM), is a soluble protein with classical superantigenic properties and is produced by an organism that causes an acute and chronic proliferative arthritis. Unfortunately, the process of obtaining purified MAM from M. arthritidis culture supernatants is extremely time-consuming and costly, and very little material is recovered. Thus, our laboratory has expressed MAM in Escherichia coli by using a protein fusion expression system. The construction and expression of recombinant MAM (rMAM), as well as a comparison of the biological properties of rMAM to those of native MAM, are discussed. Briefly, conversion of the three UGA codons to UGG codons was required to obtain full length expression and mitogenic activity of rMAM. Antisera to native MAM recognized both rMAM and the fusion protein. The T-cell receptor Vbeta and major histocompatibility complex class II receptor usages by rMAM and the fusion protein were identical to that of native MAM. In addition, the ability to induce suppression and form the superantigen bridge could also be demonstrated with rMAM. Importantly, dose-response experiments indicated that homogeneous native MAM and rMAM were of equal potency. Thus, MAM has been successfully expressed in E. coli, thereby creating a viable alternative to native MAM. PMID- 9393784 TI - Identification and characterization of a two-component regulatory system involved in invasion of eukaryotic cells and heavy-metal resistance in Burkholderia pseudomallei. AB - Burkholderia pseudomallei is the causative agent of melioidosis, a disease increasingly recognized as an important cause of morbidity and mortality in many regions of the world. B. pseudomallei is a facultative intracellular pathogen capable of invading eukaryotic cells. We used Tn5-OT182 mutagenesis to generate mutants deficient in the ability to invade a human type II pneumocyte cell line (A549 cells). One of these mutants, AJ1D8, exhibited approximately 10% of the ability of the parental strain, 1026b, to invade A549 cells. There was no difference in the abilities of 1026b and AJ1D8 to resist killing by RAW macrophages or the human defensin HNP-1. The nucleotide sequence flanking the Tn5 OT182 integration in AJ1D8 was determined, and two open reading frames were identified. The predicted proteins shared considerable homology with two component regulatory systems involved in the regulation of heavy-metal resistance in other organisms. AJ1D8 was 16-fold more sensitive to Cd2+ and twofold more sensitive to Zn2+ than was 1026b but was not sensitive to any of the other heavy metals examined. The B. pseudomallei two-component regulatory system, termed irlRS, complemented the invasion-deficient and heavy-metal-sensitive phenotype of AJ1D8 in trans. There was no significant difference between the virulence of AJ1D8 and that of 1026b in infant diabetic rats and Syrian hamsters, suggesting that the irlRS locus is probably not a virulence determinant in these animal models of acute B. pseudomallei infection. PMID- 9393786 TI - Production of Vibrio cholerae accessory cholera enterotoxin (Ace) in the yeast Pichia pastoris. AB - Accessory cholera enterotoxin (Ace) is a recently identified toxin of Vibrio cholerae. Preliminary studies using crude toxin extracts in animal models indicate that Ace increases transcellular ion transport, which is proposed to contribute to diarrhea in cholera. The lack of purified toxin has hindered elucidation of the mechanism of action of Ace. In this study, ace was cloned and was expressed in and secreted by the methylotrophic yeast Pichia pastoris. Secreted toxin constituted 50% of the total supernatant protein from Pichia pastoris. Presumed monomer and dimer forms with molecular masses of 9 and 18 kDa, respectively, were observed. The 18-kDa form predominated. Biological activity was assayed by studying ion fluxes across epithelial membranes in Ussing chambers. Among the characteristics of Ace was the unusual property of staining with silver but not Coomassie blue stain. To our knowledge this is the first report of a biologically active bacterial toxin produced with the P. pastoris system. The purified protein may now be used in studies of the mechanism of action of Ace in physiologic systems. PMID- 9393785 TI - Progression of visceral leishmaniasis due to Leishmania infantum in BALB/c mice is markedly slowed by prior infection with Trichinella spiralis. AB - We investigated in BALB/c mice the influence of the immunological environment created by the nematode Trichinella spiralis on the course of visceral leishmaniasis due to Leishmania infantum. On the day of Leishmania inoculation (day 0), mice, T. spiralis infected 7 days earlier, presented increased gamma interferon (IFN-gamma), interleukin-4 (IL-4), and IL-5 mRNA levels locally and systemically and increased the potential of spleen cells to synthesize IFN-gamma and IL-4 after activation in vitro. Eighteen days after Leishmania inoculation (day 18), corresponding to the acute phase of leishmaniasis, the hepatic amastigote burden in mice coinfected with L. infantum and T. spiralis (LT mice) was significantly lower (P < 0.001) than that in mice infected with L. infantum only (L mice). IFN-gamma and IL-4 mRNAs were overexpressed in livers of LT and L mice. On day 70, corresponding to the chronic phase, the splenic amastigote load was significantly lower (P = 0.004) in LT mice than it was in L mice. Splenic IFN gamma transcripts were overexpressed in both L and LT mice. After Leishmania specific in vitro stimulation, cytokine production was enhanced in both groups, but spleen cells from L mice produced significantly more IFN-gamma than did spleen cells from LT mice. Our data (i) generalize previous results indicating the lack of a clear-cut correlation between the outcome of murine visceral leishmaniasis and the type of cytokine pattern and (ii) demonstrate that in LT mice, leishmaniasis takes a markedly milder course than it does in L mice, providing information on the potential consequences of coinfection in a mammalian host. PMID- 9393787 TI - Isolation, cloning, and expression of a 70-kilodalton plasminogen binding protein of Borrelia burgdorferi. AB - Surface receptors for plasminogen are expressed by many gram-positive and gram negative bacteria and may play a role in the dissemination of organisms by binding plasminogen, which upon conversion to plasmin can digest extracellular matrix proteins. Two plasminogen binding proteins have been identified for Borrelia burgdorferi, outer surface protein A and a 70-kDa protein (BPBP). We purified BPBP by plasminogen affinity chromatography and obtained its amino acid sequence by Edman degradation of a tryptic digest. The gene coding for BPBP was isolated from a lambda-ZAP II genomic library with probes developed from sequenced portions of the protein. This gene was expressed in Escherichia coli; the recombinant product was seen by antibody raised against native BPBP and also bound 125I-labeled plasminogen. The experimentally derived amino acid sequences corresponded to the predicted sequence encoded by the BPBP gene. The deduced amino acid sequence for BPBP revealed significant similarity to p30, a 30-kDa protein of B. burgdorferi (54% identity and 65% similarity), to a 60-kDa protein in Borrelia coriaceae (66% identity and 80% similarity), to oligopeptide binding protein A of E. coli (34% identity and 57% similarity), and, more generally, to the periplasmic oligopeptide binding family of proteins. PMID- 9393788 TI - Therapeutic intragastric vaccination against Helicobacter pylori in mice eradicates an otherwise chronic infection and confers protection against reinfection. AB - Chronic infection of the gastroduodenal mucosae by the gram-negative spiral bacterium Helicobacter pylori is responsible for chronic active gastritis, peptic ulcers, and gastric cancers such as adenocarcinoma and low-grade gastric B-cell lymphoma. The success of eradication by antibiotic therapy is being rapidly hampered by the increasing occurrence of antibiotic-resistant strains. An attractive alternative approach to combat this infection is represented by the therapeutic use of vaccines. In the present work, we have exploited the mouse model of persistent infection by mouse-adapted H. pylori strains that we have developed to assess the feasibility of the therapeutic use of vaccines against infection. We report that an otherwise chronic H. pylori infection in mice can be successfully eradicated by intragastric vaccination with H. pylori antigens such as recombinant VacA and CagA, which were administered together with a genetically detoxified mutant of the heat-labile enterotoxin of Escherichia coli (referred to as LTK63), in which the serine in position 63 was replaced by a lysine. Moreover, we show that therapeutic vaccination confers efficacious protection against reinfection. These results represent strong evidence of the feasibility of therapeutic use of VacA- or CagA-based vaccine formulations against H. pylori infection in an animal model and give substantial preclinical support to the application of this kind of approach in human clinical trials. PMID- 9393789 TI - Transient control of interleukin-4-producing natural killer T cells in the livers of Listeria monocytogenes-infected mice by interleukin-12. AB - Unconstrained development of gamma interferon (IFN-gamma)-secreting natural killer (NK) cells and T helper (Th) 1 cells is central to protection against Listeria monocytogenes. In contrast, interleukin 4 (IL-4) is considered harmful. IL-12 produced by infected macrophages promotes, and IL-4 interferes with, protective antilisterial immunity. The liver NK T lymphocytes, which are a potent source of IL-4, are downregulated at an intermediate stage of listeriosis. Here we demonstrate that endogenous IL-12 participates in the control of IL-4 producing liver NK T lymphocytes during listeriosis. The effects of L. monocytogenes infection on IL-4-producing liver NK T lymphocytes were reversed by antibody neutralization of IL-12 but not of IFN-gamma or tumor necrosis factor alpha (TNF-alpha). IL-4 production by liver NK T lymphocytes was virtually unaffected by heat-killed L. monocytogenes (HKL). Viable L. monocytogenes markedly increased the numbers of IL-12 producers in livers in parallel with an increase in macrophage numbers, whereas HKL failed to do so with similar efficiency. These results indicate that in the liver endogenous IL-12 improves protective immunity against listeriosis by downregulating IL-4-producing NK T lymphocytes. Moreover, our findings that HKL have a low level of IL-12-inducing activity and fail to control IL-4-producing NK T lymphocytes in the liver are consistent with the lesser protective capacity of HKL compared to that of live listeriae. PMID- 9393790 TI - The acylated form of protein D of Haemophilus influenzae is more immunogenic than the nonacylated form and elicits an adjuvant effect when it is used as a carrier conjugated to polyribosyl ribitol phosphate. AB - The nonacylated form of protein D (PDm) of Haemophilus influenzae has been shown to induce the production of antibodies that are bactericidal to homologous and heterologous nontypeable H. influenzae (NTHi) strains. In this study, immunization of rats with lipoprotein D (LPD) induced higher levels of anti protein D immunoglobulin G and A serum antibodies than immunization with PDm, and the bactericidal activities of sera from LPD-immunized rats were greater than those of sera from PDm-immunized rats. Immunization with LPD or PDm did not prevent the development of acute otitis media (AOM) when rats were challenged with 10(4) CFU of an NTHi strain. However, on the eighth day of bacterial challenge, 50% (5 of 10) of LPD-immunized rats had recovered from otitis media and 30% (3 of 10) had negative middle ear cultures, whereas only 30% (3 of 10) of PDm-immunized rats had recovered, though none was culture positive. Immunization with an inactivated homologous bacterial strain elicited 70% protection (i.e., 7 of 10 rats) in the rat otitis media model. LPD and PDm were also conjugated to the H. influenzae type b (Hib) capsular polysaccharide, polyribosyl ribitol phosphate (PRP), to test protein D-conjugated PRP vaccine's potential for protection against Hib infection. When two LPD-conjugated and two PDm-conjugated PRP vaccines, each containing a different protein concentration, and a tetanus toxoid-conjugated vaccine (ACT-HIB) were tested in the experimental model of rat otitis induced with a Hib strain (Minn A), both of the LPD-conjugated and one of the PDm-conjugated vaccines induced significant protection from AOM, the level of protection being highest in animals given the vaccine with the highest LPD content. Sera from these rats also manifested the highest anti-PRP and anti-LPD antibody levels and the highest bactericidal activities against a Hib strain and an NTHi strain. PMID- 9393791 TI - Variable number of tandem repeats in clinical strains of Haemophilus influenzae. AB - An algorithm capable of identifying short repeat motifs was developed and used to screen the whole genome sequence available for Haemophilus influenzae, since some of these repeats have been shown to affect bacterial virulence. Various di- to hexanucleotide repeats were identified, confirming and extending previous findings on the existence of variable-number-of-tandem-repeat loci (VNTRs). Repeats with units of 7 or 8 nucleotides were not encountered. For all of the 3- to 6-nucleotide repeats in the H. influenzae chromosome, PCR tests capable of detecting allelic polymorphisms were designed. Fourteen of 18 of the potential VNTRs were indeed highly polymorphic when different strains were screened. Two of the potential VNTRs appeared to be short and homogeneous in length; another one may be specific for the H. influenzae Rd strain only. One of the primer sets generated fingerprint-type DNA banding patterns. The various repeat types differed with respect to intrinsic stability as well. It was noted for separate colonies derived from a single clinical specimen or strains passaged for several weeks on chocolate agar plates that the lengths of the VNTRs did not change. When several strains from different patients infected during an outbreak of lung disease were analyzed, increased but limited variation was encountered in all VNTR sites analyzed. One of the 5-nucleotide VNTRs proved to be hypervariable. This variability may reflect the molecular basis of a mechanism used by H. influenzae bacteria to successfully colonize and infect different human individuals. PMID- 9393792 TI - Natural proteoglycan receptor analogs determine the dynamics of Opa adhesin mediated gonococcal infection of Chang epithelial cells. AB - Many bacterial pathogens possess a complex machinery for the induction and/or secretion of factors that promote their uptake by mammalian cells. We searched for the molecular basis of the 60- to 90-min lag time in the interaction of Neisseria gonorrhoeae carrying the heparin-binding Opa adhesin with Chang epithelial cells. Infection assays in the presence of chloramphenicol demonstrated that the Opa-mediated gonococcal infection of Chang cells required bacterial protein synthesis when the microorganisms were derived from GC agar but not when grown in liquid media. Further analysis indicated that contact with agar ingredients rather than the growth state of the microorganisms determined the infection dynamics. DEAE chromatography of GC agar extracts and sodium dodecyl sulfate-polyacrylamide gel electrophoresis analyses and testing of collected fractions in infection assays identified negatively charged high-molecular-weight polysaccharides in the agar as inhibitors of the cellular infection. Electron microscopy showed that agar-grown gonococci were surrounded by a coat of alcian blue-positive material, probably representing accreted polysaccharides. Similar antiphagocytic material was isolated from bovine serum, indicating that in biological fluids gonococci producing the heparin-binding Opa adhesin may become covered with externally derived polysaccharides as well. Binding assays with gonococci and epithelial proteoglycan receptors revealed that polysaccharides derived from agar or serum compete with the proteoglycans for binding of the heparin-binding Opa adhesin and thus act as receptor analogs. Growth of gonococci in a polysaccharide-free environment resulted in optimal proteoglycan receptor binding and rapid bacterial entry into Chang cells. The recognition that gonococci with certain phenotypes can recruit surface polysaccharides that determine in vitro infection dynamics adds a different dimension to the well recognized biological significance of genetic variation for this pathogen. PMID- 9393793 TI - Structural and antigenic types of cell wall polysaccharides from viridans group streptococci with receptors for oral actinomyces and streptococcal lectins. AB - Lectin-mediated interactions between oral viridans group streptococci and actinomyces may play an important role in microbial colonization of the tooth surface. The presence of two host-like motifs, either GalNAc beta1-->3Gal (Gn) or Gal beta1-->3GalNAc (G), in the cell wall polysaccharides of five streptococcal strains accounts for the lactose-sensitive coaggregations of these bacteria with Actinomyces naeslundii. Three streptococcal strains which have Gn-containing polysaccharides also participate in GalNAc-sensitive coaggregations with strains of Streptococcus gordonii and S. sanguis. Each Gn- or G-containing polysaccharide is composed of a distinct phosphodiester-linked hexa- or heptasaccharide repeating unit. The occurrence of these polysaccharides on 19 additional viridans group streptococcal strains that participate in lactose-sensitive coaggregations with actinomyces was examined. Negatively charged polysaccharides that reacted with Bauhinia purpurea agglutinin, a Gal and GalNAc binding plant lectin, were isolated from 17 strains by anion exchange column chromatography of mutanolysin cell wall digests. Results from nuclear magnetic resonance and immunodiffusion identified each of 16 polysaccharides as a known Gn- or G-containing structural type and one polysaccharide as a new but closely related Gn-containing type. Unlike the reactions of lectins, the cross-reactions of most rabbit antisera with these polysaccharides were correlated with structural features other than the host-like motifs. Gn-containing polysaccharides occurred primarily on the strains of S. sanguis and S. oralis while G-containing polysaccharides were more common among the strains of S. gordonii and S. mitis examined. The findings strongly support the hypothesis that lectin-mediated recognition of these streptococci by other oral bacteria depends on a family of antigenically diverse Gn- and G containing cell wall polysaccharides, the occurrence of which may differ between streptococcal species. PMID- 9393794 TI - A specific cell surface antigen of Streptococcus gordonii is associated with bacterial hemagglutination and adhesion to alpha2-3-linked sialic acid-containing receptors. AB - A Ca2+-independent lectin activity for alpha2-3-linked sialic acid-containing receptors is associated with Streptococcus gordonii DL1 (Challis) but not with a spontaneous mutant, strain D102, that specifically lacks hemagglutinating activity. Comparison of crossed-immunoelectrophoresis patterns of parent and mutant sonicated cell extracts identified a unique antigen (Hs antigen) in the parent cell extract that was purified by DEAE Sephacel column chromatography and by a wheat germ agglutinin (WGA) lectin affinity column. The purified antigen formed a single arc in crossed immunoelectrophoresis with anti-DL1 serum and migrated as a diffuse band above the 200-kDa marker in sodium dodecyl sulfate polyacrylamide gel electrophoresis. Immunoelectron microscopy with specific anti Hs antibody revealed labeling of structures in the fibrillar layer of strain DL1 and no labeling of fibrillar structures on strain D102. Rabbit anti-DL1 serum and anti-Hs Fab inhibited the hemagglutinating activity of strain DL1, and the inhibition was specifically neutralized by purified Hs antigen. Anti-Hs Fab did not inhibit the hemagglutinating activities of several heterologous S. gordonii strains; however, these bacteria were agglutinated by anti-Hs immunoglobulin G and also by WGA. In contrast, two S. gordonii strains that lacked hemagglutinating activity did not react with anti-Hs antibody or with WGA. These findings associate the sialic acid-binding lectin activity of S. gordonii DL1 with a specific fibrillar antigen, which is composed of protein and WGA reactive carbohydrate, and indicate that cross-reactive antigens occur on other strains of this species that possess hemagglutinating activity. PMID- 9393795 TI - Protection against Pneumocystis carinii pneumonia by antibodies generated from either T helper 1 or T helper 2 responses. AB - To determine whether different antibody isotypes associated with T helper 1 (Th1) or Th2 responses are protective against Pneumocystis carinii, mice with disrupted interleukin 4 genes (IL-4(-/-) mice) or gamma interferon genes (IFN-gamma(-/-) mice) along with wild-type C57BL/6 mice were immunized intratracheally against P. carinii, depleted of T cells in vivo by use of monoclonal antibodies, and rechallenged intratracheally with 10(7) viable P. carinii organisms. Nearly all immunized mice resolved their lung P. carinii infections (limit of detection, log10 4.06) within 21 days of challenge even though they were depleted of T cells. Unimmunized mice depleted of T cells had significant lung infections (>log10 5.5) at day 21 post-P. carinii challenge. IFN-gamma(-/-) and wild-type mice developed P. carinii-specific immunoglobulin primarily of the immunoglobulin G1 (IgG1) subclass with relatively little P. carinii-specific IgG2a, IgG2b, or IgG3 in their sera, characteristic of a Th2-type response. In contrast, IL-4(-/-) mice had primarily an IgG2b P. carinii-specific antibody response in their sera with very little IgG1. Although IgG2b was the predominant isotype in IL-4(-/-) mice, optical density values of IgG2a and IgG3 were significantly higher in these mice (two and three times, respectively) than in IFN-gamma(-/-) mice, characteristic of a Th1-type response. Together, these data indicate that resolution of P. carinii infection can be mediated by specific antibody responses and that the antibody response can be either a predominantly Th1 or Th2 type. Furthermore, although wild-type mice mounted a Th2-like antibody response, IL-4( /-) mice could resolve P. carinii pneumonia, indicating that resistance to P. carinii can occur in the absence of IL-4. PMID- 9393796 TI - Characterization of a new region required for macrophage killing by Legionella pneumophila. AB - In a previous study, a collection of 55 Legionella pneumophila mutants defective for macrophage killing was isolated by transposon mutagenesis. In this study, nine of these mutants that belong to the same DNA hybridization group (group 3) were characterized. A wild-type DNA fragment that covers this DNA hybridization group was cloned and sequenced. This region was found to contain six new genes (designated icmT, icmS, icmR, icmQ, icmP, and icmO), five of which contain at least one transposon insertion. No transposon insertion was found in icmS. However, this gene was found to be required for macrophage killing, since a kanamycin resistance cassette introduced into icmS by gene replacement resulted in a mutant that was attenuated for macrophage killing. A plasmid containing the DNA fragment that covers this region complements all the mutants for macrophage killing, although various levels of complementation were observed for mutants in different genes. Complementation tests were also performed with plasmids containing one or two of these genes, as well as with plasmids containing nonpolar in-frame deletions. The results from these complementation tests indicated that all six genes located in this region are needed for macrophage killing and that they are probably arranged as two transcriptional units (icmTS and icmPO) and two genes (icmR and icmQ). A region upstream of the coding sequence of several icm genes may contain a potential promoter and/or regulatory site. Homology searches show that icmP and icmO bear significant homology to the trbA and trbC genes from the Salmonella R64 plasmid, respectively. The sequences of the other four genes do not show significant homology with any entries in sequence databases. PMID- 9393797 TI - Cryptosporidium parvum infection of human intestinal epithelial cells induces the polarized secretion of C-X-C chemokines. AB - Cryptosporidium parvum infects intestinal epithelial cells and does not invade deeper layers of the intestinal mucosa. Nonetheless, an inflammatory cell infiltrate that consists of neutrophils and mononuclear cells is often present in the lamina propria, which underlies the epithelium. This study investigated the host epithelial cell response to C. parvum by assessing in vitro and in vivo the expression and production of proinflammatory cytokines by intestinal epithelial cells after infection. The human colon epithelial cell lines HCT-8 and Caco-2 and human intestinal xenografts in SCID mice were infected with C. parvum. The expression and secretion of the C-X-C chemokines interleukin-8 (IL-8) and GROalpha were determined by reverse transcription-PCR analysis and enzyme-linked immunosorbent assay. Our results demonstrate that upregulated expression and secretion of IL-8 and GROalpha after C. parvum infection of intestinal epithelial cells first occurred 16 to 24 h after infection and increased over the ensuing 1 to 2 days. The kinetics of C-X-C chemokine production by C. parvum-infected epithelial cells contrast markedly with the rapid but transient expression of C-X C chemokines by epithelial cells infected with invasive enteric bacteria. C-X-C chemokine secretion in C. parvum-infected epithelial cells occurred predominantly from the basolateral surface in polarized monolayers of Caco-2 cells grown in Transwell cultures, whereas cell lysis occurred at the apical surface. The basolateral secretion of IL-8 and GROalpha from C. parvum-infected epithelial cells suggests that C-X-C chemokines produced by those cells contribute to the mucosal inflammatory cell infiltrate in the underlying intestinal mucosa. PMID- 9393798 TI - Invasion of brain microvascular endothelial cells by group B streptococci. AB - Group B streptococci (GBS) are the leading cause of meningitis in newborns. Although meningitis develops following bacteremia, the precise mechanism or mechanisms whereby GBS leave the bloodstream and gain access to the central nervous system (CNS) are not known. We hypothesized that GBS produce meningitis because of a unique capacity to invade human brain microvascular endothelial cells (BMEC), the single-cell layer which constitutes the blood-brain barrier. In order to test this hypothesis, we developed an in vitro model with BMEC isolated from a human, immortalized by simian virus 40 transformation, and propagated in tissue culture monolayers. GBS invasion of BMEC monolayers was demonstrated by electron microscopy. Intracellular GBS were found within membrane-bound vacuoles, suggesting the organism induced its own endocytic uptake. GBS invasion of BMEC was quantified with a gentamicin protection assay. Serotype III strains, which account for the majority of CNS isolates, invaded BMEC more efficiently than strains from other common GBS serotypes. GBS survived within BMEC for up to 20 h without significant intracellular replication. GBS invasion of BMEC required active bacterial DNA, RNA, and protein synthesis, as well as microfilament and microtubule elements of the eukaryotic cytoskeleton. The polysaccharide capsule of GBS attenuated the invasive ability of the organism. At high bacterial densities, GBS invasion of BMEC was accompanied by evidence of cellular injury; this cytotoxicity was correlated to beta-hemolysin production by the bacterium. Finally, GBS demonstrated transcytosis across intact, polar BMEC monolayers grown on Transwell membranes. GBS invasion of BMEC may be a primary step in the pathogenesis of meningitis, allowing bacteria access to the CNS by transcytosis or by injury and disruption of the endothelial blood-brain barrier. PMID- 9393799 TI - T-cell immunity to peptide epitopes of liver-stage antigen 1 in an area of Papua New Guinea in which malaria is holoendemic. AB - Liver-stage antigen 1 (LSA1) is one of several pre-erythrocytic antigens considered for inclusion in a multiantigen, multistage subunit vaccine against falciparum malaria. We examined T-cell proliferation and cytokine responses to peptides corresponding to amino acids 84 to 107, 1813 to 1835, and 1888 to 1909 of LSA1 in asymptomatic adults living in an area of Papua New Guinea where malaria is holoendemic. Whereas T cells from North Americans never exposed to malaria did not respond to any of the peptides, those from 52 of 55 adults from the area where malaria is endemic had vigorous proliferation responses to one or more of the LSA1 peptides (mean stimulation indices of 6.8 to 7.2). Gamma interferon (IFN-gamma) production driven by LSA1 peptides ranged from 34 to more than 3,500 pg/2 x 10(6) cells, was derived primarily from CD8+ cells, and was dissociated from T-cell proliferation. The frequencies of IFN-gamma response to the amino acid 1819 to 1835 and 1888 to 1909 peptides were significantly greater than that to the amino acid 84 to 107 peptide (87 and 88% versus 33% of subjects; P < 0.0001). In contrast to proliferation and IFN-gamma, interleukin 4 (IL-4) and/or IL-5 responses to LSA1 peptides were detected in only 18% of the subjects. These data show that T-cell immunity to epitopes in the N- and C-terminal regions of LSA1 are common in persons living in this area of Papua New Guinea where malaria is endemic. The dominance of type 1 CD8 cell IFN-gamma responses is consistent with a role for this T-cell population in immunity to liver-stage Plasmodium falciparum in humans. PMID- 9393800 TI - Escherichia coli cytolethal distending toxin blocks the HeLa cell cycle at the G2/M transition by preventing cdc2 protein kinase dephosphorylation and activation. AB - Cytolethal distending toxins (CDT) constitute an emerging heterogeneous family of bacterial toxins whose common biological property is to inhibit the proliferation of cells in culture by blocking their cycle at G2/M phase. In this study, we investigated the molecular mechanisms underlying the block caused by CDT from Escherichia coli on synchronized HeLa cell cultures. To this end, we studied specifically the behavior of the two subunits of the complex that determines entry into mitosis, i.e., cyclin B1, the regulatory unit, and cdc2 protein kinase, the catalytic unit. We thus demonstrate that CDT causes cell accumulation in G2 and not in M, that it does not slow the progression of cells through S phase, and that it does not affect the normal increase of cyclin B1 from late S to G2. On the other hand, we show that CDT inhibits the kinase activity of cdc2 by preventing its dephosphorylation, an event which, in normal cells, triggers mitosis. This inhibitory activity was demonstrated for the three partially related CDTs so far described for E. coli. Moreover, we provide evidence that cells exposed to CDT during G2 and M phases are blocked only at the subsequent G2 phase. This observation means that the toxin triggers a mechanism of cell arrest that is initiated in S phase and therefore possibly related to the DNA damage checkpoint system. PMID- 9393801 TI - Virulence characteristics of oral treponemes in a murine model. AB - This study was designed to investigate the virulence characteristics of Treponema denticola, T. socranskii, T. pectinovorum, and T. vincentii following challenge infection of mice. These microorganisms induced well-demarcated, dose-dependent, raised subcutaneous (s.c.) abscesses which were similar in time of onset, lesion progression, and duration of healing. Only viable cells were capable of inducing these characteristic s.c. abscesses. Histological examination of the skin lesion 3 and 5 days postinfection revealed abscess formation in the s.c. tissues, and abundant spiral organisms were demonstrated to be present in the abscess. Host resistance modulation by dexamethasone (neutrophil alteration) and cyclophosphamide (neutrophil depletion) pretreatment had a minimal effect on the virulence expression by any of these treponemes. The T. denticola isolates demonstrated significant trypsin-like protease (TLPase) activity, while both T. socranskii and T. vincentii were devoid of this activity. Interestingly, T. pectinovorum strains were heterogeneous with respect to TLPase as high producers, low producers, and nonproducers. However, no differences in lesion formation were noted regardless of whether the species expressed this proteolytic activity or whether treatment with N alpha-p-tosyl-L-lysine chloromethyl ketone (TLCK) and dithiothreitol was performed. These results showed that (i) a murine model may be used to evaluate virulence expression by oral treponemes; (ii) while TLPase activity varies among the oral treponemes, this protease does not appear to participate in abscess induction in the mouse model; and (iii) T. pectinovorum strains show variation in TLPase activity. PMID- 9393802 TI - Oligoclonality of serum immunoglobulin G antibody responses to Streptococcus pneumoniae capsular polysaccharide serotypes 6B, 14, and 23F. AB - Serum antibodies (Abs) specific for the capsular polysaccharides of Streptococcus pneumoniae provide protection against invasive pneumococcal disease. Previous studies indicate that Abs to pneumococcal polysaccharide (PPS) serotypes 1 and 6B have limited clonal diversity. To determine if restricted diversity was a feature common to other PPS specificities, we examined the light (L)-chain expression and isoelectric heterogeneity of type 6B, 14, and 23F Abs elicited in 15 adults following PPS vaccination. At the population level, both PPS-6B and PPS-14 Abs expressed kappa and lambda chains, although 6B Abs more frequently expressed lambda chains lambda and 14 Abs more frequently expressed kappa chains. In individual sera, Abs were generally skewed towards either kappa or lambda expression. 23F-specific Abs had predominantly kappa chains. Isoelectric focusing analyses showed that sera contained one or at most a few immunoglobulin G Ab spectrotypes to all three respective capsular serotypes, a result indicative of oligoclonality. A sequence analysis of a purified PPS-14-specific Ab having a single spectrotype gave uniform amino-terminal sequences for both the heavy chain (V(H)III subgroup) and the L chain (kappaIII-A27 V region). From these results we conclude that within individual adults, serum Ab responses to PPS serotypes 6B, 14, and 23F derive from a small number of dominant B-cell clones, and consequently variable-region expression is probably individually limited as well. Oligoclonality appears to be a general characteristic of human PPS-specific Ab repertoires, and we suggest that this property could lead to individual differences in Ab fine specificity and/or functional activity against encapsulated pneumococci. PMID- 9393804 TI - A mutation of F47 to A in staphylococcus enterotoxin A activates the T-cell receptor Vbeta repertoire in vivo. AB - The bacterial superantigen staphylococcal enterotoxin A (SEA) binds with high affinity to major histocompatibility complex (MHC) class II molecules and subsequently activates T cells bearing particular T-cell receptor (TCR) Vbeta chains. Structural and mutational studies have defined two distinct MHC class II binding sites located in the N-terminal and C-terminal domains of SEA. The N terminal F47 amino acid is critically involved in a low-affinity interaction to the MHC class II alpha-chain, while the C-terminal residues H187, H225, and D227 coordinate a Zn2+ ion and bind with moderate affinity to the beta-chain. In order to analyze whether the SEA-MHC class II alpha-chain interaction plays a role in dictating the in vivo repertoire of T-cell subsets, we studied distinct Vbeta populations after stimulation with wild-type SEA [SEA(wt)] and SEA with an F47A mutation [SEA(F47A)]. Injections of SEA(wt) in C57BL/6 mice induced cytokine release in serum, strong cytotoxic T-lymphocyte activity, expansion of T-cell subsets, and modulated expression of the T-cell activation antigens CD25, CD11a, CD44, CD62L, and CD69. SEA-reactive TCR Vbeta3+ and Vbeta11+ T cells were activated, while TCR Vbeta8+ T cells remained unaffected. The SEA(F47A) mutant protein induced a weaker T-cell response and failed to induce substantial interleukin-6 production compared to SEA(wt). Notably, SEA(F47A) failed to activate TCR Vbeta11+ T cells, whereas in vivo expansion and modulation of T-cell activation markers on TCR Vbeta3+ T cells were similar to those for SEA(wt). A similar response to SEA(F47A) was seen among CD4+ and CD8+ T cells. Activation of TCR Vbeta3+ and TCR Vbeta11+ T-cell hybridomas confirmed that SEA(F47A) activates TCR Vbeta3+ but not TCR Vbeta11+ T cells. The data support the view that the SEA N-terminal MHC class II alpha-chain interaction defines a topology that is required for engagement of certain TCR Vbeta chains in vivo. PMID- 9393803 TI - Isolation and characterization of a pigmentless-conidium mutant of Aspergillus fumigatus with altered conidial surface and reduced virulence. AB - Aspergillus fumigatus is an important pathogen of immunocompromised hosts, causing pneumonia and invasive disseminated disease with high mortality. The factors contributing to the predominance of A. fumigatus as an opportunistic pathogen are largely unknown. Since the survival of conidia in the host is a prerequisite for establishing disease, we have been attempting to identify factors which are associated with conidia and, simultaneously, important for infection. Therefore, an A. fumigatus mutant strain (white [W]) lacking conidial pigmentation was isolated. Scanning electron microscopy revealed that conidia of the W mutant also differed in their surface morphology from those of the wild type (WT). Mutant (W) and WT conidia were compared with respect to their capacities to stimulate an oxidative response in human phagocytes, their intracellular survival in human monocytes, and virulence in a murine animal model. Luminol-dependent chemiluminescence was 10-fold higher when human neutrophils or monocytes were challenged with W conidia compared with WT conidia. Furthermore, mutant conidia were more susceptible to killing by oxidants in vitro and were more efficiently damaged by human monocytes in vitro than WT conidia. In a murine animal model, the W mutant strain showed reduced virulence compared with the WT. A reversion analysis of the W mutant demonstrated that all phenotypes associated with the W mutant, i.e., altered conidial surface, amount of reactive oxygen species release, susceptibility to hydrogen peroxide, and reduced virulence in an murine animal model, coreverted in revertants which had regained the ability to produce green spores. This finding strongly suggests that the A. fumigatus mutant described here carries a single mutation which caused all of the observed phenotypes. Our results suggest that the conidium pigment or a structural feature related to it contributes to fungal resistance against host defense mechanisms in A. fumigatus infections. PMID- 9393805 TI - Prohibitin, a putative negative control element present in Pneumocystis carinii. AB - Little is known about the molecules involved in the regulation of Pneumocystis carinii replication and development in vitro and in vivo. We describe in this report the identification of a P. carinii gene encoding the P. carinii prohibitin protein. In mammals, the prohibitin gene product has been shown to negatively regulate cell proliferation. A cDNA clone encoding the P. carinii prohibitin gene was isolated from a P. carinii cDNA library and identified on the basis of amino acid sequence homology with prohibitin from mammalian sources. Southern blot analysis confirmed that the prohibitin cDNA clone was of P. carinii origin. Western blot analysis of total P. carinii protein indicated that the prohibitin gene is transcribed and translated in vivo. The P. carinii prohibitin gene was expressed in vivo in human fibroblasts and shown to arrest the cell cycle in the G1 phase. The results obtained suggest a potential role for P. carinii prohibitin in the regulation of P. carinii proliferation and development. PMID- 9393806 TI - Induction of neutrophil chemoattractant cytokines by Mycoplasma hominis in alveolar type II cells. AB - Bronchopulmonary dysplasia (BPD) is a chronic lung disease of premature infants who are mechanically ventilated due to respiratory distress. The disease consists of an initial inflammatory influx of neutrophils to the lungs, followed by long term chronic fibrosis of the lung tissue. The antigenic repertoire that initiates the inflammatory component of BPD has not been defined. Furthermore, the repertoire of cytokines responsible for attracting neutrophils to the lung and the mediators of pathogenesis in BPD have not been characterized. Mycoplasmas such as Mycoplasma hominis and Ureaplasma urealyticum have been isolated from the lungs of infants that developed BPD and yet have not been widely recognized as potential initiators of the inflammatory component of BPD. In the studies described here, we examined the ability of both viable and heat-killed Mycoplasma hominis to elicit type II epithelial cell production of cytokines that are chemotactic for polymorphonuclear leukocytes (PMNs), particularly interleukin-8 (IL-8) and epithelial cell-derived neutrophil-activating peptide (ENA-78). The results of these studies demonstrate that M. hominis and M. hominis antigen are potent stimulators of type II epithelial cell-derived IL-8 and ENA-78. Thus, these data strongly suggest that the presence of M. hominis in the lungs of premature infants may initiate the inflammatory component of BPD by inducing epithelial cell production of cytokines chemotactic for PMNs. Furthermore, these data suggest that the onset of the inflammatory component of BPD likely precedes, and is independent of, the recruitment and activation of alveolar macrophages. PMID- 9393807 TI - Internalin B promotes the replication of Listeria monocytogenes in mouse hepatocytes. AB - The uptake of Listeria monocytogenes by a variety of cell types in vitro is facilitated by the protein products of the inlAB (internalin) operon expressed by the organism. In the case of mouse hepatocytes, the extent to which inlAB expression influenced the uptake of Listeria in vitro was markedly dependent upon the ratio of bacteria to cells. At a ratio of 100:1, greater than 40-fold fewer transposon-induced inl4B mutant listeriae entered hepatocytes compared to the isogenic wild-type control; the difference was only fourfold, however, in cultures inoculated at a 1:1 ratio. Similarly, the uptake of in-frame inlB or inlAB deletion mutants differed only fourfold from the uptake of wild-type or inlA mutant Listeria at a 1:1 multiplicity of infection. Mutations affecting inlB or inlAB, on the other hand, resulted in a marked decrease in the capacity of Listeria to proliferate within mouse hepatocytes in vivo and in vitro. Electron micrographs of Listeria-infected hepatocytes demonstrated the impaired capacity of inlB mutants to escape from endocytic vacuoles and to enter the cytoplasm where proliferation occurs. These findings indicate that the protein product of inlB exerts a significant effect on the intracellular replication of Listeria. PMID- 9393808 TI - Contact-dependent disruption of the host cell membrane skeleton induced by Trichomonas vaginalis. AB - This report presents evidence showing that the pathogenetic process of the protozoan parasite Trichomonas vaginalis involves degradation of the target cell membrane skeleton; spectrin, the most representative protein within this structure, has been identified as the main molecular target. Degradation of the target cell spectrin is accomplished only upon contact with the parasite, and immunochemical and immunofluorescence studies performed with the erythrocyte as a model demonstrate that degradation of the protein takes place before target cell lysis. A preliminary characterization of the effectors involved has led to the identification of a nonsecreted 30-kDa proteinase which is characterized by a high specificity for spectrin. This molecule is suggested as the main effector responsible for cytoskeletal disruption. PMID- 9393809 TI - Induction of gamma interferon production in human alveolar macrophages by Mycobacterium tuberculosis. AB - Gamma interferon (IFN-gamma) is a cytokine which plays a critical role in resistance to Mycobacterium tuberculosis infection. While T lymphocytes and natural killer cells are a major source of IFN-gamma, previous demonstrations that it can be produced by murine macrophages prompted us to examine the capacity of human alveolar macrophages to express IFN-gamma. Here we report that in vitro infection of alveolar macrophages with M. tuberculosis induces both the release of IFN-gamma protein and a transient increase in IFN-gamma mRNA levels. The IFN producing cells were shown to be macrophages by reverse transcription-in situ PCR. We also observed that M. tuberculosis stimulation resulted in IFN-gamma dependent expression of the chemokines IFN-gamma-inducible protein 10 and monokine induced by IFN-gamma, suggesting that macrophage-derived IFN-gamma can function in an autocrine and/or paracrine manner. The existence of a positive regulatory loop was suggested by the observation that exogenous IFN-gamma protein could induce IFN-gamma mRNA expression in uninfected alveolar macrophages. Interleukin-12 was also found to be a potent inducer of IFN-gamma production, and M. tuberculosis-induced IFN-gamma production appears to be mediated, at least in part, by IL-12. In contrast, M. tuberculosis-induced IFN-gamma production by alveolar macrophages could be blocked by exogenous interleukin-10. These studies are the first to demonstrate an autoregulatory role for IFN-gamma produced by alveolar macrophages infected in vitro with M. tuberculosis. PMID- 9393810 TI - Invasion of dentinal tubules by oral streptococci is associated with collagen recognition mediated by the antigen I/II family of polypeptides. AB - Cell surface proteins SspA and SspB in Streptococcus gordonii and SpaP in Streptococcus mutans are members of the antigen I/II family of polypeptides produced by oral streptococci. These proteins are adhesins and mediate species specific binding of cells to a variety of host and bacterial receptors. Here we show that antigen I/II polypeptides are involved in the attachment of oral streptococci to collagen and that they also determine the ability of these bacteria to invade human root dentinal tubules. Wild-type S. gordonii DL1 (Challis) cells showed heavy invasion of tubules to a depth of approximately 200 microm, whereas the abilities of cells of isogenic mutant strains OB220 (sspA) and OB219 (sspA sspB) to invade were 50 and >90% reduced, respectively. Likewise, wild-type S. mutans NG8 cells invaded dentinal tubules, whereas cells of isogenic mutant strain 834 (spaP) did not. The invasive abilities of strains OB220 and OB219 were restored by heterologous expression of S. mutans SpaP polypeptide in these strains. The extents of tubule invasion by various wild-type and mutant strains correlated with their levels of adhesion to type I collagen, a major component of dentin. Furthermore, S. gordonii DL1 cells exhibited a growth response to collagen by forming long chains. This was not shown by ssp mutants but was restored by the expression of SpaP in these cells. The production of SspA polypeptide by S. gordonii DL1, but not production of SspB polypeptide by strain OB220 (sspA), was enhanced in the presence of collagen. These results are the first to demonstrate that antigen I/II family polypeptides bind collagen and mediate a morphological growth response of streptococci to collagen. These antigen I/II polypeptide activities are critical for intratubular growth of streptococci and thus for establishment of endodontic infections. PMID- 9393812 TI - Passive protection by polyclonal antibodies against Bacillus anthracis infection in guinea pigs. AB - The protective effects of polyclonal antisera produced by injecting guinea pigs with protective antigen (PA), the chemical anthrax vaccine AVA, or Sterne spore vaccine, as well as those of toxin-neutralizing monoclonal antibodies (MAbs) produced against PA, lethal factor, and edema factor, were examined in animals infected with Bacillus anthracis spores. Only the anti-PA polyclonal serum significantly protected the guinea pigs from death, with 67% of infected animals surviving. Although none of the MAbs was protective, one PA MAb caused a significant delay in time to death. Our findings demonstrate that antibodies produced against only PA can provide passive protection against anthrax infection in guinea pigs. PMID- 9393811 TI - The interleukin 1beta-converting enzyme, caspase 1, is activated during Shigella flexneri-induced apoptosis in human monocyte-derived macrophages. AB - Shigella, the etiological agent of bacillary dysentery, rapidly kills human monocyte-derived macrophages in vitro. Wild-type Shigella flexneri, but not a nonvirulent derivative, induced human macrophage apoptosis as determined by morphology and terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL). Shigella-mediated macrophage cell death was blocked by the peptide inhibitors of caspases, acetyl-Tyr-Val-Ala-Asp-aldehyde (acetyl-YVAD-CHO) and acetyl-Tyr-Val-Ala-Asp-chloromethylketone (acetyl-YVAD-CMK). Protection from apoptosis by YVAD was observed in monocytes matured in the presence or absence of colony-stimulating factors (CSF) like macrophage-CSF or granulocyte-macrophage CSF. Furthermore, lipopolysaccharide (LPS) or gamma interferon (IFN-gamma) rendered human macrophages partially resistant to Shigella cytotoxicity. Macrophages stimulated with either LPS or IFN-gamma were also protected by YVAD from Shigella-induced cell death. During Shigella infections of human macrophages, interleukin-1beta (IL-1beta) was cleaved to the mature form. IL 1beta maturation was severely retarded by YVAD, indicating that IL-1beta converting enzyme (ICE; caspase 1) is activated in Shigella-induced apoptosis. The finding that Shigella induces apoptosis in human macrophages by activating ICE supports the hypothesis that the acute inflammation characteristic of shigellosis is initially triggered by apoptotic macrophages which release mature IL-1beta during programmed cell death. PMID- 9393813 TI - Altered cytokine production by cystic fibrosis tracheal gland serous cells. AB - Human submucosal tracheal glands are now believed to play a major role in the physiopathology of cystic fibrosis (CF). We successfully developed techniques for culturing human tracheal gland serous cells from normal individuals (HTGS cells) and from CF patients (CF-HTGS cells) and have shown that the cultured cells have retained most of their in vivo epithelial and secretory characteristics. In order to determine to what extent the serous cells may participate in the lung defense against infection, we examined the effects of the lipopolysaccharide (LPS) of Pseudomonas aeruginosa on HTGS and CF-HTGS cells, with special reference to tumor necrosis factor alpha (TNF-alpha), interleukin-6 (IL-6), and IL-8 secretion. HTGS cells showed a daily basal secretion of IL-6 (1.68 +/- 0.14 ng/10(6) cells) and IL-8 (9.6 +/- 1.3 ng/10(6) cells) and no constitutive secretion of TNF-alpha. Treatment with P. aeruginosa LPS resulted in a significant increase in the basal production of IL-6 (increase of 200% +/- 12%) and IL-8 (525% +/- 40%) as well as a rapid production of TNF-alpha (250 +/- 38 pg/10(6) cells). The LPS-induced secretion of IL-6 and IL-8, but not that of TNF-alpha, was inhibited by glucocorticoids. CF-HTGS cells showed a much higher basal secretion of IL-6 (13.2 +/- 0.5 ng/10(6) cells) and IL-8 (45.6 +/- 7.2 ng/10(6) cells) than normal cells. Treatment with the LPS of P. aeruginosa induced increased production of IL-6 (increase of 100% +/- 8%) and IL-8 (55% +/- 18%) but did not induce the secretion of TNF-alpha. Neither intracellular TNF-alpha nor TNF-alpha transcripts were found in CF-HTGS cells, whereas they were found in normal HTGS cells. In addition, dexamethasone was found to stimulate IL-6 and IL-8 secretion (in the presence or absence of LPS) but did not induce any secretion of TNF-alpha. All these data indicate that HTGS cells are responsive to P. aeruginosa LPS, which results in an increased secretion of IL-6, IL-8, and TNF-alpha, the secretion of which appeared to be impaired in CF-HTGS cells. PMID- 9393814 TI - Human B- and T-cell responses after immunization with a hexavalent PorA meningococcal outer membrane vesicle vaccine. AB - The PorA protein from Neisseria meningitidis, a potential vaccine candidate, induces human bactericidal antibodies which are serosubtype specific. We developed a hexavalent PorA outer membrane vesicle vaccine based on reference strain H44/76. This vaccine contains the six most prevalent PorA serosubtypes as found in many countries. We previously reported on the immune responses of 20 adult volunteers after a single immunization with this vaccine. In this study, the B- and T-cell responses in three adult volunteers were studied after three consecutive immunizations (0, 2, and 11 months). The first immunization induced a strong B-cell response resulting in high immunoglobulin G levels in an outer membrane vesicle enzyme-linked immunosorbent assay. At least a fourfold increase in bactericidal activity was observed against the majority (four to six) of the vaccine antigens compared to prevaccination titers. Immunodominance was observed for one or two of the PorAs in the bactericidal assay with a set of six isogenic H44/76-derived PorA target strains. These strains carry the individual PorAs as present in the vaccine. The second and third immunizations did not induce a further increase in the immune responses. A decline in time with respect to PorA specific antibodies was observed after each immunization. These observations were reflected by the T-cell proliferation responses. Two additional sets of isogenic H44/76-derived mutant strains were used to study the specificity and/or cross reactivity of the induced bactericidal antibodies. These target strains differ only in expressing mutant family variants of either PorA P1.7,16 or P1.5,10, both present in the PorA vesicle vaccine. The bactericidal antibody responses found were directed predominantly against the P1.7 (loop 1 of P1.7,16) and the P1.10 (loop 4 of P1.5,10) epitopes. This indicates that different portions of PorA were involved in the induction of bactericidal antibodies depending upon the PorA serosubtype. PMID- 9393815 TI - Systemic infection of mice by wild-type but not Spv- Salmonella typhimurium is enhanced by neutralization of gamma interferon and tumor necrosis factor alpha. AB - The spv genes of the virulence plasmid of Salmonella typhimurium and other nontyphoidal serovars of S. enterica are involved in systemic infection by increasing the replication rate of the bacteria in host tissues beyond the intestines. We considered the possibility that the Spv virulence function is to evade suppression by the host response to infection. To examine this possibility, gamma interferon (IFN-gamma) and/or tumor necrosis factor alpha (TNF-alpha) were neutralized in BALB/c mice by intraperitoneal administration of monoclonal antibodies. Neutralization of IFN-gamma and/or TNF-alpha resulted in increased splenic infection with wild-type salmonellae after oral inoculation; however, Spv salmonellae were defective at increasing splenic infection in cytokine-depleted mice. The use of a temperature-sensitive marker plasmid, pHSG422, indicated that neutralization of IFN-gamma caused less killing of wild-type S. typhimurium, while neutralization of TNF-alpha resulted in an increased in vivo replication rate for wild-type salmonellae. These results demonstrate that the Spv virulence function is not to evade suppression of bacterial infection normally mediated by IFN-gamma or TNF-alpha. PMID- 9393817 TI - Differential induction of Th1 versus Th2 cytokines by group A streptococcal toxic shock syndrome isolates. AB - The majority of group A streptococcal (GAS) isolates from patients with streptococcal toxic shock syndrome (STSS) and necrotizing fasciitis (NF) express numerous virulence factors, including several superantigens (SAgs). Purified SAgs are potent inducers of inflammatory (Th1) cytokines that contribute to the pathogenesis of severe infections. However, GAS-infected individuals are likely to be exposed to a mixture of GAS SAgs as well as other virulence factors produced by the bacteria, and therefore, our goal was to characterize the mitogenic and cytokine induction profiles of this mixture. All GAS isolates tested had brisk mitogenic activity and induced potent cytokine responses, with higher frequencies of Th1 than Th2 cytokine-producing cells. The mitogenic activity produced in culture supernatants of three selected clinical GAS isolates was significantly different, but no marked difference was found in their overall cytokine induction profiles. However, significant differences (P < 0.0062) were noted in the induction of Th2 cytokines between GAS supernatants and recombinant streptococcal pyrogenic exotoxin A (rSpeA), suggesting that the presence of other SAgs and/or the production of additional virulence factors may alter the overall cytokine induction profile of SAgs. A significant individual variation in the level of proliferative and cytokine responses to the same GAS culture supernatants or to rSpeA was noted. Individuals with higher frequencies of cells producing Th2 cytokines mounted lower levels of Th1 cytokine responses, and vice versa. Furthermore, quantification of the intensity and cell area of interleukin 1beta (IL-1beta)-producing cells by image analysis revealed that individuals with higher Th2 responses had significantly lower IL-1beta production (P < 0.0001) than the individual with a strong Th1 response. Differences in the ability to induce Th1 versus Th2 cytokines, as well as the individual variations in cytokine responses to streptococcal SAgs, may play a central role in determining the severity of invasive GAS infections. PMID- 9393816 TI - Identification of homing receptors that mediate the recruitment of CD4 T cells to the genital tract following intravaginal infection with Chlamydia trachomatis. AB - Murine genital infection induced with the mouse pneumonitis biovar of Chlamydia trachomatis (MoPn) elicits a short-lived protective immunity mediated primarily by Th1 CD4 cells. To understand the development of local cell-mediated immunity against C. trachomatis infection, we investigated the mechanism(s) which mediates CD4 lymphocyte migration to the genital mucosa by identifying molecules that could support this process. We found that primarily CD4 cells were recruited to the genital tract (GT) during primary and challenge MoPn infection. Peak levels were found 21 days after primary inoculation (15.4% +/- 2.7%) and 7 days (31.3% +/- 8.5%) after challenge but diminished after resolution of infection. The CD4 cells appeared to be recruited to the GT in response to infection since these cells expressed the profile of activated, or memory, cells. We also observed up regulation of homing receptors containing LFA-1 (CD11a) and alpha4 (CD49d) on GT CD4 cells over the course of infection. Furthermore, the mucosal homing receptor chain, beta7, but not the peripheral homing receptor chain beta1 (CD29), was detected on GT CD4 cells. MoPn-infected GT tissue expressed the endothelial cell ligands vascular cell adhesion molecule 1 (VCAM-1), intracellular adhesion molecule 1 (ICAM-1), and mucosal vascular addressin cell adhesion molecule 1 (MAdCAM-1), which correspond to the homing receptors on GT CD4 cells. Interestingly, VCAM-1 and MAdCAM-1 were not expressed in the GTs of uninfected mice but were temporarily induced following infection, indicating that expression of endothelial ligands in the GT are regulated by chlamydial infection. These data suggest that recruitment of CD4 cells to the GT is mediated through LFA 1:ICAM-1 and alpha4beta7:MAdCAM-1-VCAM-1 interactions. PMID- 9393818 TI - Immunogenicity and protective efficacy of the alpha C protein of group B streptococci are inversely related to the number of repeats. AB - Infection by group B streptococci (GBS) is an important cause of bacterial disease in neonates. Alpha C protein is a protective cell surface-associated protein of GBS. This protein contains a repeat region flanked by N and C termini. Variable expression of tandem repeating units of alpha C proteins had been found among clinical isolates of GBS. We examined the effect of the number of repeats on the immunogenicity of the alpha C protein and its ability to elicit protection from GBS infection in a neonatal mouse model. Mice were immunized with purified alpha C proteins of constructs containing various numbers of repeats (n = 1, 2, 9, and 16) and the N- and C-terminal regions. Both the N-terminal and the repeat regions contain protective and opsonic epitopes. Antibody responses to the alpha C protein constructs with various numbers of repeats were tested with enzyme linked immunosorbent assay plates coated with either native, nine-repeat alpha C protein or "repeatless" N-terminal antigen. An inverse relationship was found between the number of repeats and the immunogenicity of the alpha C protein; this effect was most pronounced on titers of antibody to the N-terminal region. An inverse relationship was also observed between the number of repeats and protective efficacy, i.e., mouse dams immunized with 5 microg of one- or nine repeat alpha C protein transferred protective immunity to 65 or 11% of their pups, respectively (P < 0.0001). Thus, the presence of multiple repeats appears to lessen the antibody response to the complete alpha C protein, and especially the antibody response to its N-terminal region, and suggests a mechanism whereby repeat elements contribute to the evasion of host immunity. PMID- 9393820 TI - Identification of a Fusobacterium nucleatum PK1594 galactose-binding adhesin which mediates coaggregation with periopathogenic bacteria and hemagglutination. AB - Attachment of Fusobacterium nucleatum to various oral surfaces is mediated by several adhesins anchored on its outer surface. Monoclonal antibodies (MAbs) were prepared and used to identify the putative galactose-binding adhesin of F. nucleatum PK1594. Four unique MAbs, 8G7, 26B9, 28G11, and 29D4, were isolated on the basis of their ability to inhibit coaggregation of F. nucleatum PK1594 with Porphyromonas gingivalis PK1924. All four MAbs were also capable of inhibiting galactose-inhibitable interactions of F. nucleatum PK1594 with other oral gram negative bacteria and with erythrocytes. Preincubation of F. nucleatum PK1594 with MAb 26B9 or its Fab fragments at concentrations lower than 1 microg/ml resulted in complete inhibition of coaggregation with P. gingivalis PK1924 or hemagglutination. F. nucleatum PK1594 surface components prepared by mild sonication or by extracting whole cells with detergents were subjected to Western blot analysis. None of the MAbs were able to recognize any polypeptide in these experiments. Therefore, detergent extracts of F. nucleatum PK1594 surface components were subjected to three experimental procedures: (i) separation by ion exchange chromatography and testing of fractions for reaction with MAb 26B9 in an enzyme-linked immunosorbent assay (ELISA), (ii) lactose-Sepharose affinity chromatography and testing of the lactose eluate in ELISA with MAb 26B9, and (iii) immunoseparation with either MAb 26B9 or 8G7. Collectively, the results suggest that the putative adhesin is a 30-kDa outer membrane polypeptide which mediates the coaggregation with P. gingivalis PK1924 as well as other galactose sensitive interactions of F. nucleatum PK1594. PMID- 9393819 TI - Identification and characterization of a K88- and CS31A-like operon of a rabbit enteropathogenic Escherichia coli strain which encodes fimbriae involved in the colonization of rabbit intestine. AB - Initiation of attaching-effacing lesions, which characterize infections with rabbit enteropathogenic Escherichia coli (REPEC), requires bacteria to adhere to the intestinal epithelium. This adherence is reflected in vitro by the affinity of these E. coli strains for various types of eukaryotic cells. TnphoA mutants of REPEC 83/39 (O15:H-) which had lost the ability to adhere to HEp-2 epithelial cells, guinea pig ileal brush borders, and mouse erythrocytes were generated. DNA sequencing of the region surrounding the inactivating transposon insertions within a 95-kb plasmid, designated pRAP for REPEC adherence plasmid, revealed extensive homology between that region and the structural genes of enterotoxigenic E. coli operons encoding the K88 and CS31A fimbrial adhesins and the genes for the afr2 adhesin from REPEC B10 (O103:H2). Seven genes of the ral operon (for REPEC adherence locus), including three putative minor fimbrial subunit genes (ralC, ralF, and ralH), a major fimbrial subunit gene (ralG), a gene of unknown function (ralI), and genes for two fimbrial subunit chaperones (ralD and ralE), were sequenced. When inoculated perorally into weanling rabbits, a mutant with a TnphoA insertion in the ralE gene showed a 10-fold reduction in colonizing ability, with only 1 of 10 rabbits excreting bacteria compared to all 5 of those infected with the wild-type parent strain (P = 0.002). The severity of the diarrheal illness caused by the mutant strain was also reduced. Western blotting of surface protein extracts of strain 83/39 with hyperimmune anti-83/39 antiserum, adsorbed with the ralE mutant, revealed a 32-kDa protein which was absent from protein extracts of two nonadherent mutants. The adsorbed antiserum also bound to the surface of strain 83/39 but not to nonadherent mutants, as detected by immunogold labeling. These results indicate that the ral operon of REPEC 83/39 contains genes necessary for the biosynthesis of fine fimbriae which are responsible for in vitro adherence of the bacteria and play a role in their colonization of, and hence virulence for, rabbits. The putative major fimbrial subunit is a protein with an observed molecular size of approximately 32 kDa which, when assembled, appears to form a capsule of fimbriae surrounding the bacterium similar to that described for CS31A. PMID- 9393821 TI - Pathogenicity and protective effect of rough mutants of Salmonella species in germ-free piglets. AB - In this study, two stable, rough, streptomycin-sensitive Salmonella mutants with different types of genetic defects were used to colonize groups of germ-free (GF) piglets. The lipopolysaccharide (LPS) of Salmonella typhimurium SF 1591 was of the Ra chemotype (complete core), whereas the LPS of the S. minnesota mR 595 deep rough mutant contained only lipid A and 2-keto-3-deoxyoctulosonic acid (Re chemotype). Both strains readily colonized the intestinal tracts of GF piglets and were stable during the whole experiment. All animals survived, and only transient fever was observed in some piglets colonized with the SF 1591 strain. Finally, streptomycin and virulent, smooth, streptomycin-resistant S. typhimurium LT2 were administered perorally 1 week later. All piglets colonized previously with the deep-rough mutant mR 595 died of sepsis, in contrast to piglets infected with the LT2 strain and colonized with the SF 1591 mutant, all of which survived. This difference is explained by the penetration of the mesenteric lymph nodes, spleen, and liver by great numbers of live bacteria in the latter case, resulting in prominent systemic and local immune responses. On the other hand, live bacteria were found only rarely in the mesenteric lymph nodes of animals colonized with the mR 595 strain and a negligible antibody response was observed. PMID- 9393822 TI - Cellular changes and cytokine expression in the ilea of gnotobiotic piglets resulting from peroral Salmonella typhimurium challenge. AB - Two stable rough mutants of Salmonella spp. were studied as live peroral vaccines. The SF1591 mutant of S. typhimurium (Ra chemotype) protected germ-free piglets against subsequent infection with virulent smooth S. typhimurium LT2, whereas a deep-rough mutant of S. minnesota mR595 (Re chemotype) did not. We investigated cytokine and leukocyte profiles in the ilea of gnotobiotic piglets colonized for 1 week either with rough mutants alone or with rough mutants followed by S. typhimurium LT2. The ileal mucosae of piglets associated with strain SF1591 alone were not inflamed. Villi contained activated macrophages, and enterocytes expressed transforming growth factor beta (TGF-beta). Subsequent infection of piglets with S. typhimurium LT2 resulted in immigration of alphabeta T cells and immunoglobulin A (IgA) response. In contrast, the ileal mucosae of piglets associated with strain mR595 alone expressed heat shock proteins and inflammatory cytokines but not TGF-beta. Acellular villi contained numerous gammadelta T cells but no alphabeta T cells. After subsequent challenge with the LT2 strain, most piglets died of sepsis. Intestinal mucosae contained IgG but no IgA. These findings suggest the importance of cytokine signals in the regulation of intestinal responses against Salmonella infection. PMID- 9393823 TI - Mapping of antigenic determinant regions of the Bor56 protein of Orientia tsutsugamushi. AB - The 56-kDa protein (Bor56) of Orientia tsutsugamushi is an immunoprotective antigen and is the target molecule of neutralizing antibodies. This antigen is recognized by almost all of the serum antibodies produced by patients in the convalescence phase of scrub typhus. We expressed the Bor56 open reading frame in Escherichia coli and generated from it a series of deletion constructs as MalE fusion proteins. Antibody-binding domains were characterized by using patient sera, mouse monoclonal antibodies (MAbs), and Bor56-immunized-mouse sera. None of the antibodies bound to a fusion protein containing the carboxy-terminal 140 amino acids (aa) of the Bor56 protein, suggesting that the carboxy-terminal domain of Bor56 is not exposed on the surface of the molecule. Human immunoglobulin M (IgM) antibodies predominantly bound to antigenic domain I (AD I; amino acids [aa] 19 to 113) and AD III (aa 243 to 328). Human IgG antibodies also showed preferential binding to AD I. The epitope recognized by strain specific MAb (KI4) or group-specific MAb (KI57) was mapped to AD II (aa 142 to 203). Mouse serum antibodies, elicited by immunization with deletion mutants, consistently bound to AD III. Moreover, the carboxy-terminal 140 aa of the Bor56 protein did not elicit an antibody response in C3H/HeDub mice. A model of the antigenic structure of Bor56 is presented and discussed. These results suggest that antigenic fragments from AD I and AD III are useful in the induction of humoral immunity against O. tsutsugamushi. These antigenic analyses provide an important foundation for further analyses of the neutralizing-antibody responses generated during rickettsial infections. They also provide potential peptide substrates for diagnostic assays and vaccine strategies. PMID- 9393824 TI - Calcium dependence and binding in cultures of Histoplasma capsulatum. AB - Histoplasma capsulatum is a pathogenic fungus with two distinct morphologies and lifestyles. The saprophytic form of this organism, a mold, thrives in soil and is especially abundant in the Ohio and Mississippi River valleys. Its parasitic counterpart, a yeast, colonizes phagolysosomes of mammalian macrophages. We have observed a major difference in the calcium requirements of the two forms of Histoplasma, potentially implicating the phagolysosome as a calcium-limiting compartment. Deprivation of calcium by the addition of EGTA to culture media inhibited the growth of mycelial H. capsulatum but had no effect on yeast growth in vitro. In addition, yeasts released a calcium-binding protein (CBP) detectable by a 45CaCl2 blotting technique. CBP was a major component of yeast culture supernatant and was also detectable by ruthenium red staining, another assay for calcium-binding activity. Conversely, mycelial H. capsulatum did not produce CBP, a finding that correlates with the dependence of mycelia on calcium for growth. We also describe here the purification of CBP from yeast culture supernatant by reversed-phase high-pressure liquid chromatography. PMID- 9393825 TI - Apoptosis of human monocytes and macrophages by Mycobacterium avium sonicate. AB - Mycobacterium avium is an intracellular organism which multiplies predominantly within human macrophages. This organism has previously been shown to induce apoptosis in human macrophages. With a view to identifying M. avium components that induce cell death in infected host cells, sonicated extracts of M. avium as well as individual components isolated from the M. avium sonicate were tested in various assays with a human monocytic cell line (THP-1). THP-1 cells incubated with M. avium sonicate showed significantly reduced viability after a 2-day exposure compared to control cells incubated with media alone. This effect was dose dependent, with only 6.6% +/- 5.2% and 48.8% +/- 10.3% of the cells being viable by trypan blue exclusion at 600 and 300 microg/ml, respectively. Control cells, on the other hand, exhibited a viability of 98.8% +/- 1.0%. In addition, an 80% ammonium sulfate fraction of the M. avium sonicate and the previously characterized 68-kDa protein were found to have similar effects on THP-1 cells. In both cases, the reduction in viability was due to apoptosis characterized by chromatin condensation, DNA fragmentation by agarose gel electrophoresis, or terminal deoxynucleotidyl transferase-mediated d-UTP nick end labeling (TUNEL) and release of nuclear matrix protein (NMP) into the culture medium. M. avium sonicate-induced apoptosis of THP-1 cells was completely inhibited by the commonly used antioxidants pyrrolidinedithiocarbamate (PDTC) and butylated hydroxyanisole (BHA), indicating that the generation of free oxygen radicals may be responsible for inducing cell death. M. avium sonicate was found to induce apoptosis of monocyte-derived macrophages (MDMs) as well. This effect was not reversed in the presence of PDTC and was not accompanied with DNA fragmentation when determined by agarose gel electrophoresis, as seen in the case of THP-1 cells. However, these MDMs were found to contain fragmented DNA by TUNEL. These findings suggest that the mechanism of cell death in MDMs may be different from that observed with THP-1 cells. Furthermore, these results provide new insight into the effect of M. avium components on host cell responses during M. avium infection. PMID- 9393827 TI - Comparison of inducible nitric oxide synthase expression in the brains of Listeria monocytogenes-infected cattle, sheep, and goats and in macrophages stimulated in vitro. AB - The expression of inducible nitric oxide synthase (iNOS) was studied in the brains of cattle, sheep, and goat that succumbed to a natural infection with Listeria monocytogenes. The lesions in infected brains are characterized by microabscesses, perivascular cuffs, gliosis, glial nodules, and large areas of malacia. Using immunocytochemistry, we detected bacteria in microabscesses, particularly in sheep and goats, and in areas without signs of inflammation, but not in perivascular infiltrates. iNOS was expressed by macrophage (Mphi)-type cells of microabscesses and glial nodules but rarely by Mphi in areas of malacia, as determined by immunohistochemistry with iNOS-specific antibodies. iNOS was not detected in perivascular cuffs. Major histocompatibility complex class II molecules (MHC-II), another marker of cell activation, showed a different pattern of distribution. Perivascular cuffs contained high numbers of MHC-II-positive cells, including some with Mphi characteristics. Microabscesses in sheep and goats showed low expression of MHC-II, particularly in iNOS-expressing cells. In cattle, the expression of markers for activated or recruited phagocytes, the calcium-binding proteins S100A8 and S100A9 (formerly called MRP-8 and MRP-14, respectively), was largely restricted to cells showing weak or undetectable iNOS expression; iNOS-positive Mphi showed a low expression of S100A8 and S100A9. Thus, iNOS is expressed by a restricted subset of Mphi in listeric encephalitis. In cultured sheep and goat Mphi, a low proportion of cells expressed iNOS upon activation by L. monocytogenes and gamma interferon, resulting in nitrite generation at least 1 order of magnitude lower than that in similarly treated cattle Mphi. Since these species differences were much less obvious in vivo, it appears that the well-known species variation in iNOS expression by Mphi could reflect an in vitro phenomenon. PMID- 9393826 TI - Effects of mycobacteria on regulation of apoptosis in mononuclear phagocytes. AB - Since apoptosis is observed in tuberculous granulomata, we investigated the molecular mechanisms underlying the apoptotic pathway in an in vitro model of mycobacterial infection of mononuclear phagocytes. We postulated that Mycobacterium tuberculosis could trigger the apoptotic pathway in macrophages, resulting in death of the microorganism by modulating the expression of bcl-2, bax, bcl-xL, and bcl-xS. We found that the mRNA of bcl-2, an inhibitor of apoptosis, was downregulated in peripheral blood monocytes (PBM) between 2 and 6 h following infection with M. bovis BCG or induction with heat-killed M. tuberculosis H37Ra. Western analysis showed a downregulation of the Bcl-2 protein, with a half-life of 24 h. At the same time points, there was no change in the expression of Bax or Bcl-xS, inducers of apoptosis, but Bcl-xL, another inhibitor of apoptosis, was minimally upregulated by BCG. To determine if apoptosis could be a mechanism for growth inhibition in vivo, we obtained alveolar macrophages by bronchoalveolar lavage from involved sites in patients with active pulmonary tuberculosis. Using the TUNEL (terminal deoxynucleotidyltransferase mediated nick end labeling) technique, we observed significantly more apoptosis in involved segments of five tuberculosis patients (14.8 +/- 1.9%) than in those of normal controls (<1%, P = 0.02) or in uninvolved segments (4.3 +/- 0.9%, P < 0.05). We conclude that apoptosis of mononuclear phagocytes induced by M. tuberculosis occurs in vivo and that in an in vitro model of mycobacterial infection, apoptosis may be mediated by downregulation of Bcl-2. PMID- 9393828 TI - Expression, cloning, and characterization of a Candida albicans gene, ALA1, that confers adherence properties upon Saccharomyces cerevisiae for extracellular matrix proteins. AB - Adherence of Candida albicans to host tissues is a necessary step for maintenance of its commensal status and is likely a necessary step in the pathogenesis of candidiasis. The extracellular matrix (ECM) proteins are some of the host tissue and plasma proteins to which C. albicans adheres through adhesins located on the fungal cell surface. To isolate genes encoding ECM adhesins, an assay was developed based on the ability of yeast cells to adhere to magnetic beads coated with the ECM protein fibronectin, type IV collagen, or laminin. A C. albicans genomic library was constructed by cloning XbaI-partially-digested and size selected fragments into pAUR112, an Escherichia coli-yeast low-copy-number shuttle vector. The C. albicans library was transformed into Saccharomyces cerevisiae YPH 499, and clones capable of adherence were selected by using ECM protein-coated magnetic beads. A plasmid containing an approximately 8-kb insert was isolated from 29 adherent clones. These clones exhibited adherence to all ECM protein-coated magnetic beads and to human buccal epithelial cells. The ALA1 gene (for agglutinin-like adhesin) was localized by subcloning it into a 5-kb XbaI fragment which retained the adherence phenotype in both orientations. The complete DNA sequence of the 5-kb insert was determined, and an open reading frame (ORF) encoding 1,419 amino acid residues was identified. Deletions from the 5' and 3' ends extending into the DNA sequence encoding the 1,419-amino-acid ORF product inactivated the adherence phenotype, suggesting that it is the coding region of the ALA1 gene. A database search identified ALA1 to be similar to the C. albicans ALS1 (for agglutinin-like sequence 1) protein and the S. cerevisiae agglutinin protein (AG alpha1), although the homology at the primary amino acid sequence level is limited to the first half of each of these proteins. ALA1 contains a central domain of six tandem repeats of 36 amino acids. We discuss the significance of various predicted ALA1 structural motifs and their relationships to function in the adherence process. PMID- 9393829 TI - A highly adherent phenotype associated with virulent Bvg+-phase swine isolates of Bordetella bronchiseptica grown under modulating conditions. AB - The ability of Bvg(-)-phase and Bvg(+)-phase Bordetella bronchiseptica swine isolates, grown under modulating or nonmodulating conditions, to adhere to swine ciliated nasal epithelial cells was determined. When virulent strains were cultivated at 37 degrees C in the Bvg+ phase, numerous adherent bacteria (approximately eight per cell, depending on the strain used) were observed. However, when such strains were grown under modulating conditions (23 degrees C), a significant increase in the level of attachment was seen, suggesting that B. bronchiseptica produces a Bvg-repressed adhesin under these conditions. bvg mutant strains, including an isogenic bvgS mutant, adhered minimally. Western blots indicated that two putative B. bronchiseptica adhesins, filamentous hemagglutinin and pertactin, were not detectable in cultures displaying the highly adherent phenotype. Several proteins apparent in Western blots obtained by using bacterial extracts enriched in outer membrane proteins derived from B. bronchiseptica grown at 23 degrees C were not present in similar extracts prepared from an isogenic bvgS mutant grown at 23 degrees C or from the parent strain grown at 37 degrees C. Adherence of bacteria cultivated at 23 degrees C was almost completely abolished by pretreatment of organisms at 60 degrees C; adherence was reduced by 57% when bacteria were pretreated with pronase E. Temperature shift experiments revealed that the heightened level of adhesion that occurs following growth at 23 degrees C was maintained for up to 18 h when bacteria were subsequently incubated at 37 degrees C. We propose that a Bvg repressed adhesin, expressed only by modulated bvg+ strains of B. bronchiseptica, may play a key role in the initial colonization of naturally infected swine. PMID- 9393830 TI - Infection with Chlamydia trachomatis alters the tyrosine phosphorylation and/or localization of several host cell proteins including cortactin. AB - Infection of epithelial cells by two biovars of Chlamydia trachomatis results in the tyrosine phosphorylation of several host proteins. The most prominent change in host protein tyrosine phosphorylation involves a complex of proteins with molecular masses of 75 to 85 kDa (pp75/85) and 100 kDa (pp100). The C. trachomatis-induced tyrosine phosphorylation of pp75/85 and pp100 is observed in several cell lines, including epithelial cells, fibroblasts, and macrophages. Subcellular fractionation and detergent solubility properties of pp75/85 are consistent with its association with the cytoskeleton. Phosphoamino acid analysis demonstrates that the pp75/85 complex is phosphorylated on both tyrosine and serine residues. Immunofluorescence studies of chlamydia-infected cells by using fluorescein isothiocyanate-phalloidin and antibodies to phosphotyrosine and cortactin demonstrate that tyrosine-phosphorylated proteins, as well as cortactin, are localized to the chlamydial vacuole and that this process is facilitated by actin. PMID- 9393831 TI - Internalin of Listeria monocytogenes with an intact leucine-rich repeat region is sufficient to promote internalization. AB - Listeria monocytogenes can use two different surface proteins, internalin (InlA) and InlB, to invade mammalian cells. The exact role of these invasiveness factors in vivo remains to be determined. In cultured cells, InlA is necessary to promote Listeria entry into human epithelial cells, such as Caco-2 cells, whereas InlB is necessary to promote Listeria internalization in several other cell types, including hepatocytes, fibroblasts, and epithelioid cells, such as Vero, HeLa, CHO, or Hep-2 cells. We have recently reported that the InlA receptor on Caco-2 cells is the cell adhesion molecule E-cadherin and demonstrated that nonpermissive fibroblasts become permissive for internalin-mediated entry when transfected with the gene coding for LCAM, the chicken homolog of the human E cadherin gene. In this study, we demonstrate for the first time that the internalin protein alone is sufficient to promote internalization into cells expressing its receptor. Indeed, internalin confers invasiveness to both Enterococcus faecalis and internalin-coated latex beads. As shown by transmission electron microscopy, these beads were phagocytosed via a "zipper" mechanism similar to that observed during the internalin-E-cadherin-mediated entry of Listeria. Moreover, a functional analysis of internalin demonstrates that its amino-terminal region, encompassing the leucine-rich repeat (LRR) region and the inter-repeat (IR) region, is necessary and sufficient to promote bacterial entry into cells expressing its receptor. Several lines of evidence suggest that the LRR region would interact directly with E-cadherin, whereas the IR region would be required for a proper folding of the LRR region. PMID- 9393832 TI - Expression of thin aggregative fimbriae promotes interaction of Salmonella typhimurium SR-11 with mouse small intestinal epithelial cells. AB - The factors that mediate binding of Salmonella typhimurium to small intestinal epithelial cells have not been fully characterized. In this paper we demonstrate that elimination of production of thin aggregative fiber by a transposon insertion within the gene encoding the subunit protein of the fiber reduced binding of S. typhimurium SR-11 to a conditionally immortalized proximal small intestinal epithelial cell line established from transgenic mice. This binding defect could be overcome by transcomplementation with a wild-type allele. The conditionally immortalized cell line should prove useful in identifying the epithelial cell receptor for bacterial attachment since expression of its bacterial binding activity can be induced by manipulating the line's proliferative status. PMID- 9393833 TI - A nonamer peptide derived from Listeria monocytogenes metalloprotease is presented to cytolytic T lymphocytes. AB - Listeria monocytogenes is an intracellular bacterium that secretes proteins into the cytosol of infected macrophages. Major histocompatibility complex (MHC) class I molecules bind peptides that are generated by the degradation of bacterial proteins and present them to cytolytic T lymphocytes (CTL). In this study we have investigated CTL responses in L. monocytogenes-immunized mice to peptides that (i) derive from the L. monocytogenes proteins phosphatidylinositol-specific phospholipase C, lecithinase (most active on phosphatidylcholine), metalloprotease (Mpl), PrfA, and the ORF-A product and (ii) conform to the binding motif of the H2-Kd MHC class I molecule. We identified a nonamer peptide, Mpl 84-92, that is presented to L. monocytogenes-specific CTL by H2-Kd MHC class I molecules. Unlike other motif-conforming peptides derived from the secreted Mpl of L. monocytogenes, Mpl 84-92 is bound with high affinity by H2-Kd. Mpl 84-92 is the fourth L. monocytogenes-derived peptide found to be presented to CTL by the H2-Kd molecule during infection and demonstrates the importance of high-affinity interactions between antigenic peptides and MHC class I molecules for CTL priming. PMID- 9393834 TI - The role of Legionella pneumophila-infected Hartmannella vermiformis as an infectious particle in a murine model of Legionnaire's disease. AB - Legionella pneumophila is a bacterial parasite of many species of freshwater protozoa and occasionally an intracellular pathogen of humans. While protozoa are known to play a key role in the persistence of L. pneumophila in the environment, there has been limited research addressing the potential role of L. pneumophila infected protozoa in the pathogenesis of human infection. In this report, the potential role of an L. pneumophila-infected amoeba as an infectious particle in replicative L. pneumophila lung infection was investigated in vivo with the amoeba Hartmannella vermiformis, a natural reservoir of L. pneumophila in the environment. L. pneumophila-infected H. vermiformis organisms were prepared by coculture of the amoebae and virulent L. pneumophila cells in vitro. A/J mice, which are susceptible to replicative L. pneumophila lung infection, were subsequently inoculated intratracheally with L. pneumophila-infected H. vermiformis organisms (10(6) amoebae containing 10(5) bacteria), and intrapulmonary growth of the bacteria was assessed. A/J mice inoculated intratracheally with L. pneumophila-infected H. vermiformis organisms developed replicative L. pneumophila lung infections. Furthermore, L. pneumophila-infected H. vermiformis organisms were more pathogenic than an equivalent number of bacteria or a coinoculum of L. pneumophila cells and uninfected amoebae. These results demonstrate that L. pneumophila-infected amoebae are infectious particles in replicative L. pneumophila infections in vivo and support the hypothesis that inhaled protozoa may serve as cofactors in the pathogenesis of pulmonary disease induced by inhaled respiratory pathogens. PMID- 9393835 TI - CD8+ T cells are activated during the early Th1 and Th2 immune responses in a murine Lyme disease model. AB - T-helper responses following Borrelia burgdorferi infection in mice determine susceptibility to Lyme arthritis. The ratio of interleukin 4-positive, CD4+ to gamma interferon (IFN-gamma)-positive, CD4+ T cells was significantly greater in infected BALB/cJ mice than in infected C3H/HeJ mice. Increased numbers of IFN gamma-producing cells predicted greater arthritis severity, and CD8+ T cells were the main source of IFN-gamma in both strains. PMID- 9393836 TI - A systemic downregulation of gamma interferon production is associated with acute shigellosis. AB - Production of cytokines by peripheral blood mononuclear cells from Shigella infected patients was assessed. The frequencies of tumor necrosis factor alpha (TNF-alpha), TNF-beta, and transforming growth factor beta mRNA-expressing cells were persistently upregulated during the course of shigellosis in comparison to those of healthy controls. In contrast, the frequency of gamma interferon (IFN gamma) mRNA-expressing cells was significantly reduced during the acute stage compared to that during the convalescent stage and to that of healthy Bangladeshi controls (P < 0.01). Constitutive IFN-gamma production in Bangladeshi controls was significantly upregulated compared to that in Swedish controls. PMID- 9393837 TI - The role of complement, antibody, and tumor necrosis factor alpha in the killing of Plasmodium falciparum by the monocytic cell line THP-1. AB - Killing of Plasmodium falciparum blood forms by the differentiated human myelomonocytic THP-1Mo cell line was studied by a radiometric assay. Results showed that parasite killing was promoted by complement, antimalarial antibody, and the cytokines tumor necrosis factor alpha and gamma interferon. Differentiated THP-1Mo appears to be a useful monocytic cell line for the study of mechanisms of immunity to Plasmodium. PMID- 9393838 TI - Interference of peptides and specific antibodies with the function of the Actinobacillus pleuropneumoniae transferrin-binding protein. AB - Multiple-antigenic peptides (MAPs) containing transferrin-binding domains of the Actinobacillus pleuropneumoniae serotype 7-derived transferrin-binding protein (TfbA) (K. Strutzberg, L. von Olleschik, B. Franz, C. Pyne, M. A. Schmidt, and G. F. Gerlach, Infect. Immun. 63:3846-3850, 1995) were constructed. It was found that the MAPs inhibited transferrin binding of the recombinant TfbA protein, whereas antibodies directed against transferrin-binding domains failed to do so. PMID- 9393839 TI - Experimental infection of C3H mice with avian, porcine, or human isolates of Serpulina pilosicoli. AB - C3H/HeJ (lps(d)/lps(d)) and C3H/HeOuJ (lps(n)/lps(n)) mice were infected via gastric intubation with avian, porcine, or human isolates of weakly hemolytic spirochetes classified as Serpulina pilosicoli. Upon histopathological examination of cecal tissue from mice infected with avian or porcine isolates, colonization of spirochetes attached end-on to the apical surface of enterocytes was observed. There were no apparent differences in severity of cecal lesions between the lipopolysaccharide (LPS)-responsive (C3H/HeOuJ) and LPS hyporesponsive (C3H/HeJ) mouse strains infected with these isolates. Transmission electron microscopy showed spirochetes invaginated into the host cell membrane with resultant effacement of microvilli and loss of the glycocalyx. End-on attachment of the human isolate S. jonesii was not observed in the present studies, although weakly hemolytic spirochetes were reisolated from mice infected with S. jonesii. Moreover, results of Western immunoblot experiments showed mice developed serum antibody responses to the S. pilosicoli isolates examined. Thus, the present results indicate that specific isolates of S. pilosicoli can colonize mice and exhibit end-on attachment to cecal enterocytes. PMID- 9393840 TI - A monoclonal antibody to Candida albicans enhances mouse neutrophil candidacidal activity. AB - A monoclonal antibody (MAb) to Candida albicans (MAb B6.1) that protects against candidiasis and the nonprotective MAb B6 were compared for ability to support neutrophil (polymorphonuclear leukocyte [PMN]) candidacidal activity. Both MAbs are immunoglobulin M, and each recognizes distinct C. albicans mannan cell wall determinants. PMN candidacidal activity was assessed by transmission electron microscopy and by an in vitro killing assay. The results indicated that MAb B6.1, but not MAb B6, enhances ingestion and killing of yeast cells by PMN in the presence of serum complement. PMID- 9393841 TI - Identification of shuA, the gene encoding the heme receptor of Shigella dysenteriae, and analysis of invasion and intracellular multiplication of a shuA mutant. AB - shuA encodes a 70-kDa outer membrane heme receptor in Shigella dysenteriae. Analysis of the shuA DNA sequence indicates that this gene encodes a protein with homology to TonB-dependent receptors of gram-negative bacteria. Transport of heme by the ShuA protein requires TonB and its accessory proteins ExbB and ExbD. The shuA DNA sequence contains a putative Fur box overlapping the -10 region of a potential shuA promoter, and the expression of shuA is repressed by exogenous iron or hemin in a Fur-dependent manner, although hemin repressed expression to a lesser extent than iron salts. Disruption of this open reading frame on the S. dysenteriae chromosome by marker exchange yielded a strain that failed to use heme as an iron source, indicating that shuA is essential for heme transport in S. dysenteriae. However, shuA is not essential for invasion or multiplication within cultured Henle cells; the shuA mutant invaded and produced normal plaques in confluent cell monolayers. PMID- 9393842 TI - Characterization of lipoprotein IRP1 from Corynebacterium diphtheriae, which is regulated by the diphtheria toxin repressor (DtxR) and iron. AB - The Corynebacterium diphtheriae irp1 gene is negatively regulated by DtxR and iron. The nucleotide sequence of irp1 revealed that it has homology with genes involved in iron acquisition. Expression of the irp1 gene showed that it encodes a lipoprotein (IRP1) with a predicted size of 38 kDa. Northern blot experiments indicated that transcription from the irp1 promoter is repressed in high-iron medium and suggested that irp1 is part of an iron-regulated operon. PMID- 9393843 TI - Association of Leishmania heat shock protein 83 antigen and immunoglobulin G4 antibody titers in Brazilian patients with diffuse cutaneous leishmaniasis. AB - Diffuse cutaneous leishmaniasis (DCL) is characterized by the presence of numerous nonulcerated nodules and plaques containing large numbers of Leishmania amazonensis parasites and few lymphoid elements. The immune responses of DCL patients reflect severe antigen-specific T-cell deficiencies, while the antibody response to Leishmania antigens is often accentuated. We report herein on the Leishmania antigen-specific antibody subclass distribution in DCL patients and demonstrate that a dominant antigen contributing to the biased immunoglobulin G4 antibody subclass in sera of DCL patients is Leishmania heat shock protein 83. PMID- 9393845 TI - Quantification of conserved antigens in Helicobacter pylori during different culture conditions. AB - In this study, we raised monoclonal antibodies (MAbs) against three conserved Helicobacter pylori antigens, i.e., the N-acetylneuraminyllactose-binding fibrillar hemagglutinin, HpaA; the flagellin subunits, FlaA and FlaB; and a species-specific 26-kDa protein. The MAbs were used for the development of sensitive inhibition enzyme-linked immunosorbent assays for quantification of these antigens in H. pylori during various culture conditions. The quantities of these antigens varied considerably (up to 8-fold) during different culture procedures and between strains (up to 10-fold). PMID- 9393844 TI - The transcriptional regulator SoxS is required for resistance of Salmonella typhimurium to paraquat but not for virulence in mice. AB - In Escherichia coli, the SoxRS regulon is required for resistance to redox cycling agents which elevate cytosolic superoxide levels, as well as for resistance to nitric oxide-dependent macrophage killing. In Salmonella typhimurium, SoxS is also required for enhanced expression of Mn-superoxide dismutase and resistance to paraquat, but not for resistance to nitric oxide donor compounds in vitro, resistance to macrophage killing, or virulence in mice. Differences in other antioxidant defense systems or compensation by homologous regulons may account for species-specific differences in the role of SoxS. PMID- 9393846 TI - Characterization and immunogenicity of Salmonella typhimurium SL1344 and UK-1 delta crp and delta cdt deletion mutants. AB - S. typhimurium SL1344 and UK-1 mutants with deletions of the crp (cyclic AMP receptor protein) and cdt (colonization of deep tissues) genes have been constructed and characterized, and their levels of virulence and immunogenicity have been determined for BALB/c mice. All Crp- Cdt- and Crp+ Cdt- mutants were avirulent, as mice survived oral doses of 10(9) cells without illness. All the mutants colonized the gut-associated lymphoid tissue efficiently, but capacities to colonize deeper tissues, such as those of the spleen and liver, and blood were significantly reduced for the Crp- Cdt- and Crp+ Cdt- mutants compared with the Crp- Cdt+ mutant and the wild-type parent strain. The Crp- Cdt- and Crp+ Cdt- SL1344 strains induced complete protection, as all mice immunized with the mutants survived challenge with approximately 10(4) times the 50% lethal dose of the wild-type SL1344 strain. The Crp- UK-1 strain was least attenuated yet induced the highest level of protective immunity against challenge with the wild type UK-1 strain. The Crp+ Cdt- and Crp- Cdt- strains, although totally attenuated, differed in immunogenicity to challenge with the highly virulent UK-1 parent, with the apparently hyperattenuated Crp- Cdt- strain inducing a lower level of protective immunity than the Crp+ Cdt- strain. Nevertheless, these UK-1 Crp- Cdt- and Crp+ Cdt- strains induced complete protective immunity to challenge with the less-virulent SL1344 wild-type strain. Taken collectively, the results indicate that the attenuation of a highly virulent S. typhimurium strain can yield a vaccine that induces excellent protective immunity to challenge with less virulent S. typhimurium strains. PMID- 9393847 TI - Splicing into senescence: the curious case of p16 and p19ARF. PMID- 9393848 TI - Cytochrome c: can't live with it--can't live without it. PMID- 9393849 TI - Protein translocation in the three domains of life: variations on a theme. PMID- 9393850 TI - Biogenesis of the gram-negative bacterial envelope. PMID- 9393851 TI - Membrane protein biogenesis: regulated complexity at the endoplasmic reticulum. PMID- 9393852 TI - Impaired maternal behavior in mice lacking norepinephrine and epinephrine. AB - The roles of norepinephrine (NE) and epinephrine in behavior were investigated by targeted disruption of the dopamine beta-hydroxylase (Dbh) gene, thereby eliminating these compounds in vivo. Most heterozygous pups born to Dbh-/- females died within several days of birth and were often found scattered within the bedding. Potential causes including deficits in olfaction and lactation were not apparent. A deficit in maternal behavior was confirmed by the lack of pup retrieval exhibited by Dbh-/- virgin females. Restoration of NE shortly before but not after birth induced females that previously abandoned their litters to act maternally. Our results suggest that NE is responsible for long-lasting changes that promote maternal behavior during both development and parturition in mice. PMID- 9393853 TI - Complex polymorphisms in an approximately 330 kDa protein are linked to chloroquine-resistant P. falciparum in Southeast Asia and Africa. AB - Chloroquine resistance in a P. falciparum cross maps as a Mendelian trait to a 36 kb segment of chromosome 7. This segment harbors cg2, a gene encoding a unique approximately 330 kDa protein with complex polymorphisms. A specific set of polymorphisms in 20 chloroquine-resistant parasites from Asia and Africa, in contrast with numerous differences in 21 sensitive parasites, suggests selection of a cg2 allele originating in Indochina over 40 years ago. One chloroquine sensitive clone exhibited this allele, suggesting another resistance component. South American parasites have cg2 polymorphisms consistent with a separate origin of resistance. CG2 protein is found at the parasite periphery, a site of chloroquine transport, and in association with hemozoin of the digestive vacuole, where chloroquine inhibits heme polymerization. PMID- 9393854 TI - Acidic sphingomyelinase mediates entry of N. gonorrhoeae into nonphagocytic cells. AB - Invasion of human mucosal cells by N. gonorrhoeae via the binding to heparansulfate proteoglycan receptors is considered a crucial event of the infection. Using different human epithelial cells and primary fibroblasts, we show here an activation of the phosphatidylcholine-specific phospholipase C (PC PLC) and acidic sphingomyelinase (ASM) by N. gonorrhoeae, resulting in the release of diacylglycerol and ceramide. Genetic and/or pharmacological blockade of ASM and PC-PLC cause inhibition of cellular invasion by N. gonorrhoeae. Complementation of ASM-deficient fibroblasts from Niemann-Pick disease patients restored N. gonorrhoeae-induced signaling and entry processes. The activation of PC-PLC and ASM, therefore, is an essential requirement for the entry of N. gonorrhoeae into distinct nonphagocytic human cell types including several epithelial cells and primary fibroblasts. PMID- 9393855 TI - Regulation of Golgi structure through heterotrimeric G proteins. AB - We have previously shown that ilimaquinone (IQ), a marine sponge metabolite, causes complete vesiculation of the Golgi stacks. By reconstituting the IQ mediated vesiculation of the Golgi apparatus in permeabilized cells, we now demonstrate that this process does not require ARF and coatomers, which are necessary for the formation of Golgi-derived COPI vesicles. We find that IQ mediated Golgi vesiculation is inhibited by G alpha(s)-GDP and G alpha(i3)-GDP. Interestingly, adding betagamma subunits in the absence of IQ is sufficient to vesiculate Golgi stacks. Our findings reveal that IQ-mediated Golgi vesiculation occurs through activation of heterotrimeric G proteins and that it is the free betagamma, and not the activated alpha subunit, that triggers Golgi vesiculation. PMID- 9393856 TI - Bcl-xL regulates the membrane potential and volume homeostasis of mitochondria. AB - Mitochondrial physiology is disrupted in either apoptosis or necrosis. Here, we report that a wide variety of apoptotic and necrotic stimuli induce progressive mitochondrial swelling and outer mitochondrial membrane rupture. Discontinuity of the outer mitochondrial membrane results in cytochrome c redistribution from the intermembrane space to the cytosol followed by subsequent inner mitochondrial membrane depolarization. The mitochondrial membrane protein Bcl-xL can inhibit these changes in cells treated with apoptotic stimuli. In addition, Bcl-xL expressing cells adapt to growth factor withdrawal or staurosporine treatment by maintaining a decreased mitochondrial membrane potential. Bcl-xL expression also prevents mitochondrial swelling in response to agents that inhibit oxidative phosphorylation. These data suggest that Bcl-xL promotes cell survival by regulating the electrical and osmotic homeostasis of mitochondria. PMID- 9393857 TI - Avian hairy gene expression identifies a molecular clock linked to vertebrate segmentation and somitogenesis. AB - We have identified and characterized c-hairy1, an avian homolog of the Drosophila segmentation gene, hairy. c-hairy1 is strongly expressed in the presomitic mesoderm, where its mRNA exhibits cyclic waves of expression whose temporal periodicity corresponds to the formation time of one somite (90 min). The apparent movement of these waves is due to coordinated pulses of c-hairy1 expression, not to cell displacement along the anteroposterior axis, nor to propagation of an activating signal. Rather, the rhythmic c-hairy mRNA expression is an autonomous property of the paraxial mesoderm. These results provide molecular evidence for a developmental clock linked to segmentation and somitogenesis of the paraxial mesoderm, and support the possibility that segmentation mechanisms used by invertebrates and vertebrates have been conserved. PMID- 9393858 TI - Tumor suppression at the mouse INK4a locus mediated by the alternative reading frame product p19ARF. AB - The INK4a tumor suppressor locus encodes p16INK4a, an inhibitor of cyclin D dependent kinases, and p19ARF, an alternative reading frame protein that also blocks cell proliferation. Surprisingly, mice lacking p19ARF but expressing functional p16INK4a develop tumors early in life. Their embryo fibroblasts (MEFs) do not senesce and are transformed by oncogenic Ha-ras alone. Conversion of ARF+/+ or ARF+/- MEF strains to continuously proliferating cell lines involves loss of either p19ARF or p53. p53-mediated checkpoint control is unperturbed in ARF-null fibroblast strains, whereas p53-negative cell lines are resistant to p19ARF-induced growth arrest. Therefore, INK4a encodes growth inhibitory proteins that act upstream of the retinoblastoma protein and p53. Mutations and deletions targeting this locus in cancer cells are unlikely to be functionally equivalent. PMID- 9393859 TI - Identification of clonogenic common lymphoid progenitors in mouse bone marrow. AB - The existence of a common lymphoid progenitor that can only give rise to T cells, B cells, and natural killer (NK) cells remains controversial and constitutes an important gap in the hematopoietic lineage maps. Here, we report that the Lin( )IL-7R(+)Thy-1(-)Sca-1loc-Kit(lo) population from adult mouse bone marrow possessed a rapid lymphoid-restricted (T, B, and NK) reconstitution capacity in vivo but completely lacked myeloid differentiation potential either in vivo or in vitro. A single Lin(-)IL-7R(+)Thy-1(-)Sca-1loc-Kit(lo) cell could generate at least both T and B cells. These data provide direct evidence for the existence of common lymphoid progenitors in sites of early hematopoiesis. PMID- 9393860 TI - MAP kinases with distinct inhibitory functions impart signaling specificity during yeast differentiation. AB - Filamentous invasive growth of S. cerevisiae requires multiple elements of the mitogen-activated protein kinase (MAPK) signaling cascade that are also components of the mating pheromone response pathway. Here we show that, despite sharing several constituents, the two pathways use different MAP kinases. The Fus3 MAPK regulates mating, whereas the Kss1 MAPK regulates filamentation and invasion. Remarkably, in addition to their kinase-dependent activation functions, Kss1 and Fus3 each have a distinct kinase-independent inhibitory function. Kss1 inhibits the filamentation pathway by interacting with its target transcription factor Ste12. Fus3 has a different inhibitory activity that prevents the inappropriate activation of invasion by the pheromone response pathway. In the absence of Fus3, there is erroneous crosstalk in which mating pheromone now activates filamentation-specific gene expression using the Kss1 MAPK. PMID- 9393861 TI - The MinE ring: an FtsZ-independent cell structure required for selection of the correct division site in E. coli. AB - E. coli cell division is mediated by the FtsZ ring and associated factors. Selection of the correct division site requires the combined action of an inhibitor of FtsZ ring formation (MinCD) and of a topological specificity factor that somehow prevents MinCD action at the middle of the cell (MinE). Here we show that a biologically active MinE-Gfp fusion accumulates in an annular structure near the middle of young cells. Formation of the MinE ring required MinD but was independent of MinC and continued in nondividing cells in which FtsZ function was inhibited. The results indicate that the MinE ring represents a novel cell structure, which allows FtsZ ring formation at midcell by suppressing MinCD activity at this site. PMID- 9393862 TI - Crystal structure at 1.7 A resolution of VEGF in complex with domain 2 of the Flt 1 receptor. AB - Vascular endothelial growth factor (VEGF) is a homodimeric hormone that induces proliferation of endothelial cells through binding to the kinase domain receptor and the Fms-like tyrosine kinase receptor (Flt-1), the extracellular portions of which consist of seven immunoglobulin domains. We show that the second and third domains of Flt-1 are necessary and sufficient for binding VEGF with near-native affinity, and that domain 2 alone binds only 60-fold less tightly than wild-type. The crystal structure of the complex between VEGF and the second domain of Flt-1 shows domain 2 in a predominantly hydrophobic interaction with the "poles" of the VEGF dimer. Based on this structure and on mutational data, we present a model of VEGF bound to the first four domains of Flt-1. PMID- 9393863 TI - Metals, motifs, and recognition in the crystal structure of a 5S rRNA domain. AB - Two new RNA structures portray how non-Watson-Crick base pairs and metal ions can produce a unique RNA shape suitable for recognition by proteins. The crystal structures of a 62 nt domain of E. coli 5S ribosomal RNA and a duplex dodecamer encompassing an internal loop E have been determined at 3.0 and 1.5 A, respectively. This loop E region is distorted by three "cross-strand purine stacks" and three novel, water-mediated noncanonical base pairs and stabilized by a four metal ion zipper. These features give its minor groove a unique hydrogen bonding surface and make the adjacent major groove wide enough to permit recognition by the ribosomal protein L25, which is expected to bind to this surface. PMID- 9393864 TI - Effect of high-energy phosphates and free radical scavengers on replant survival in an ischemic extremity model. AB - In replantation surgery, preoperative and intraoperative ischemia can lead to irreversible changes that prevent reperfusion during the subsequent re establishment of circulation. These changes are termed the no-reflow phenomenon. Ischemic phase damage was addressed by comparing the dose-response effects of controls vs. five different high-energy phosphate compounds on replanted limb survival. Reperfusion damage was evaluated via comparisons of controls with superoxide dismutase (SOD). Ischemic hindlimbs treated with high-energy phosphates displayed improved survival compared with controls. Limbs treated with SOD demonstrated no change in survival at 4 hours and improved survival at 8 hours. Combining adenosine and SOD had no improved effect on survival. Adenosine was the most effective high-energy phosphate in limiting ischemic damage. The free radical scavenger (SOD) was beneficial only at the later stages of ischemia. In this experimental model, there appears to be a role for both phosphates and free radical scavengers in enhancing ischemic tissue survival. PMID- 9393865 TI - Application of hemodilution in microsurgical free flap transplantation. AB - The intra-operative hemodilution and blood auto-transfusion is a blood-saving technique that can be performed when major blood loss is expected. The effects of this technique were studied in 30 microsurgical free flap transplantation patients. Between 400-600 ml blood was collected from the patients before surgery. The patients received dextran, a balanced salt solution, and glucose with the ratio of 3:1 to the collected blood volume, bleeding, and urine before the blood auto-transfusion. There were no significant changes in RBC, HCT, MCV, blood pressure, or heart rate. Of the 30 free flaps, 28 were successful with a 93.3% survival rate. The safety of intra-operative hemodilution and autologous blood transfusion in microsurgery as well as the effect of hemodilution on transplanted flap survival are discussed in this study. PMID- 9393866 TI - Effect of anastomosis and geometry of vessel curvature on blood flow velocity and patency in microvessels. AB - The effect of the geometry of the vessel and the number of anastomoses on the blood flow was studied. Four different shapes of the vessel were constructed by using a 6-cm-long double vein graft model with three anastomoses: (1) an alpha loop, (2) an omega loop, (3) a sigmoid curve, and (4) straight. Blood flow was measured by an ultrasound Doppler flowmeter. The result showed no alternation in blood flow across different geometry and through three patent microanastomoses. However, six out of seven vein grafts were thrombosed at 24 hr postoperative due to vascular kinks. This model demonstrates potential sites of kinking at the dissection end of the femoral artery, the microanastomoses, the side branches of the vein graft, and the adventitial adhesions. This model is recommended to microvascular trainees for the study of kinking and the management of redundant pedicles and vein grafting. PMID- 9393867 TI - Actions of glucocorticosteroids on ischemic-reperfused muscle and cutaneous tissue. AB - Ischemia-reperfusion injury represents a complex series of vascular and cellular events that resembles an acute inflammatory reaction within the reperfused tissue. This article provides an overview of glucocorticosteroid effects on cells and tissues involved in inflammatory reaction following ischemia-reperfusion of muscle and cutaneous tissue. Glucocorticosteroids exert a variety of effects that influence the microcirculation. These effects include leukocyte recruitment, reduction of vascular permeability, inhibition of formation of cytokines or other mediators, and modulation of enzyme systems involved in inflammation. The current view is that glucocorticosteroids act through cytoplasmic receptors by controlling the transcription of certain genes encoding regulatory proteins, but the exact mechanisms of glucocorticoid action on ischemia-reperfusion are not completely understood. Potential mechanisms may involve modulation of neutrophil and endothelial function, inhibition of formation of arachidonic acid products, and attenuation of lipid peroxidation of biological membranes through membrane stabilization and scavenging of toxic free radicals. PMID- 9393868 TI - Free posterior tibial perforator flap: anatomy and a report of 6 cases. AB - Anatomy of cutaneous perforators of the posterior tibial artery were studied in 20 limbs of 10 cadavers. The majority of the perforators (n = 74, 61%) were located in the middle two quarters of the leg, at an average of 18.6 cm (s.d. 4.5 cm; range 10.5-26 cm) from the medial malleolus, or around 54% (s.d 16%) of the length of the leg. There were usually 3 or 4 perforators in this region, with an average caliber of 1.5 mm (s.d. 0.2 mm; range 1-2 m.m.) and an average length from the posterior tibial artery to the skin of 4.0 cm (s.d. 1.3 cm; range 2.5-6 c.m.). A free fasciocutaneous skin flap based on one of these perforators (the posterior tibial perforator flap, PTP flap) was successfully transplanted in 6 cases. This modified technique of the posterior tibial flap enables the surgeon to retain the posterior tibial artery when the skin of the medial aspect of the leg is chosen to be used as skin flap donor. PMID- 9393869 TI - Effect of vein grafting on the survival of microvascularly transplanted muscle flaps. AB - Since vein grafting is often required during transplantation of free muscle flaps but is associated with a higher failure rate than those flaps transplanted with primary anastomoses, we sought to compare primary repair with the use of vein grafting in an experimental setting. We transplanted the gracilis muscle to the contralateral side in 98 rats using four different methods of vessel repair. In the Control group (n = 28), both femoral vessels were anastomosed primarily. In Experimental Group 1 (n = 25), the femoral artery was anastomosed with an epigastric vein graft and the femoral vein was anastomosed primarily. In Experimental Group 2 (n = 25), the femoral vein was anastomosed with a femoral vein graft and the femoral artery was anastomosed primarily. In Experimental Group 3 (n = 20), both femoral vessels were anastomosed with vein grafts. The Control and Experimental Groups 1-3 survival rates were 89.3, 76.0, 84.0, and 70.0%, respectively; none of the experimental group survival rates was significantly different from that of the control (P < 0.5). This study demonstrates that the use of size-matched, interpositional vein grafts in the arterial or venous pedicle of the rat gracilis muscle flap during transplantation did not significantly decrease flap survival as compared to primary arterial or venous anastomoses. If the observed failure rate persisted with expansion of the study groups to 60-100 animals each, the failure rate of flaps with vein grafts would be significantly lower and comparable to failure rates reported in some clinical series. The large numbers necessary to significantly show this decrease make this model impractical for further studies. PMID- 9393870 TI - Influence of postischemic administration of oxyradical antagonists on ischemic injury to rabbit skeletal muscle. AB - The aim of this study was to determine whether the administration of free radical antagonists, immediately before and during the early minutes of reperfusion, improves muscle survival 24 hr after a period of ischemia. Rabbit rectus femoris muscles were isolated, made ischemic for 3 1/2 hr and treated with either desferrioxamine (DFX), an Fe3+ chelator, superoxide dismutase and catalase (SOD & CAT), which quench superoxide and hydrogen peroxide, or allopurinol, an inhibitor of xanthine oxidase (XO). After 24 hr reperfusion, muscle viability (+/-s.e.m.), measured by the nitro blue tetrazolium (NBT) vital staining technique, was 41.6 +/- 11.3% for saline-treated ischemic controls, 30.6 +/- 7.6% for DFX-treated, 46.7 +/- 10.3% for SOD & CAT-treated, and 43.3 +/- 9.5% for allopurinol-treated muscles. None of the treated groups differed significantly from the ischemic control group. Tissue myeloperoxidase, ATP and reduced glutathione levels, and plasma lactate dehydrogenase (LDH) and aspartate transaminase (AST) levels were increased by ischemia and reperfusion in all groups, but the changes did not differ between the treatment groups. Levels of XO in the rabbit muscle were determined and found to be very low in both normal and postischemic muscle. As XO is the target enzyme of allopurinol, its absence provides a basis for the lack of effect of this agent. However, it is not clear why DFX and SOD & CAT had no protective effect. PMID- 9393871 TI - The multiple endocrine neoplasia type 2B point mutation switches the specificity of the Ret tyrosine kinase towards cellular substrates that are susceptible to interact with Crk and Nck. AB - The RET proto-oncogene encodes a Tyrosine Kinase Receptor (RTK) which plays an important function in the proliferation and/or differentiation of neuroectodermic cells. Germline mutation of a methionine to a threonine within the RET TK domain predisposes to the Multiple Endocrine Neoplasia type 2B (MEN 2B). It has been demonstrated that, unlike c-Ret, the MEN 2B mutated Ret displays constitutive TK activity, tyrosine autophosphorylation and transforms fibroblasts. However, this oncoprotein is more than a fully activated wild-type (WT) Ret TK since it also displays modified substrate specificity. Change in substrate specificity leads to the tyrosine autophosphorylation of MEN 2B Ret on new sites as well as the phosphorylation of several novel downstream targets. But, none of these substrates have been identified and the ability of MEN 2B Ret phosphoprotein to interact with Src Homology 2 (SH2) domain containing molecules has been poorly investigated. In this report, using a constitutively activated Ret TK form, Ret ptc 2, we demonstrate that the MEN 2B as the activated WT Ret TK binds to several SH2 signalling proteins such as Shc, Grb-2, Phospholipase Cgamma, Crk and Nck. However, in contrast to the activated WT form, expression of the MEN 2B mutated Ret-ptc 2 results in the tyrosine phosphorylation of a panel of proteins which interestingly interact with Crk and Nck. We identified Paxillin, a cytoskeletal protein as one of the Crk associated proteins that is dramatically phosphorylated in MEN 2B but not in WT Ret expressing cells. These data suggest that MEN 2B mutated Ret triggers distinct signalling pathways that might be related to its transforming power. PMID- 9393872 TI - Activation of p65 NF-kappaB protein by p210BCR-ABL in a myeloid cell line (P210BCR-ABL activates p65 NF-kappaB). AB - The chimeric tyrosine kinase p210BCR-ABL is involved in the pathogenesis of chronic myelogenous leukemia. It transforms immature hematopoietic cells in vitro and abrogates IL-3-dependent growth. The mechanisms by which p210BCR-ABL mediates its oncogenicity are not well elucidated. Identifying transcription factors targeted by the chimeric protein may help to clarify these mechanisms. We have analysed the effect of p210BCR-ABL expression on NF-kappaB activity in DA1 cells (an IL-3-dependent murine myeloid progenitor cell line). A specific stimulation of NF-kappaB activity by kinase-active wild-type p210BCR-ABL has been evidenced by transcriptional activation assays. Electrophoretic mobility supershift assays revealed the presence of p65 protein (RelA) DNA binding activity in p210BCR-ABL transformed DA1 cells but not in parental DA1 cells. Activation of RelA in transformed DA1 cells may occur by protein stabilization. Experiments using oligonucleotides antisense to RelA showed that p210BCR-ABL transfected cells failed to survive after IL-3 removal. Moreover, inhibition of cellular growth was shown following treatment of p210BCR-ABL transformed DA1 cells by p65 antisense oligonucleotides. This study suggests that p65 NF-kappaB may be an effector for p210BCR-ABL and probably contributes to its induced transformation process. PMID- 9393873 TI - JNK1, JNK2 and JNK3 are p53 N-terminal serine 34 kinases. AB - The function of the tumor suppressor protein p53 is modulated by post translational events, primarily by phosphorylation. p53 is phosphorylated at multiple sites by a variety of protein kinases depending on the cellular environment. It has been suggested that serine 34 of mouse p53 is specifically phosphorylated by a stress-activated protein kinase in response to ultraviolet radiation. Since serine 34 is a major site of phosphorylation of mouse p53 in vivo and its specific protein kinase is still not definitively identified yet, we have examined the c-Jun N-terminal kinase 1 (JNK1) activity on p53 by expressing JNK1 in 293T cells. We show here that activated JNK1 phosphorylates mouse p53 specifically at serine 34 in vitro, while a dominanant-negative JNK1 mutant does not phosphorylate p53. More importantly, JNK1 associates with p53 in vivo, with or without activation, confirming that JNK1 is indeed a p53 kinase. Interestingly, activated JNK2 and JNK3 also phosphorylate serine 34 of mouse p53. Furthermore, JNK2 and JNK3 also associate with p53 in vivo, indicating that not only JNK1, but also JNK2 and JNK3 are p53 N-terminal serine 34 kinases. Phosphorylation of p53 by JNKs may play an important role in nuclear signal transduction in response to environmental stress or tumorigenic agents. PMID- 9393874 TI - Choline kinase inhibitors as a novel approach for antiproliferative drug design. AB - Recent progress in deciphering the molecular basis of carcinogenesis is of utmost importance to the development of new anticancer strategies. To this end, it is essential to understand the regulation of both normal cell proliferation and its alterations in cancer cells. We have previously demonstrated that in ras transformed cells there is an increased level of phosphorylcholine (PCho) resulting from a constitutive activation on choiline kinase (ChoK). The importance of ChoK for the regulation of cell proliferation has also been proposed since an inhibitor for this enzyme, hemicholinium-3 (HC-3), drastically reduces entry into the S phase after stimulation with growth factors. Here we report the synthesis of several new compounds which are highly specific inhibitors for ChoK, with up to 1000-fold or 600-fold increased inhibitory activity, compared to HC-3 under ex vivo or in vitro conditions respectively. These novel compounds also drastically reduce entry into the S phase after stimulation with specific growth factors. A more profound inhibition of cell proliferation was observed in ras-, src- and mos-transformed cells in the presence of ChoK inhibitors, compared to their parental, untransformed NIH3T3 cells. By contrast, this effect was not observed in fos-transformed cells. While ras, src and mos transformation is associated with elevated levels of ChoK activity, fos-induced transformation does not affect ChoK activity. The inhibitory effect on proliferation of the new compounds correlates with their ability to inhibit the production of phosphorylcholine in whole cells, a proposed novel second messenger for cell proliferation. These results strongly support a critical role of choline kinase in the regulation of cell growth and makes this enzyme a novel target for the design of new antiproliferative and anticancer drugs. PMID- 9393875 TI - AU-rich mRNA instability elements on human papillomavirus type 1 late mRNAs and c fos mRNAs interact with the same cellular factors. AB - Expression levels of human papillomavirus type 1 late genes are determined in part by an AU-rich inhibitory sequence in the 3' untranslated region on the late mRNAs. Fine mapping of the AU rich inhibitory sequence revealed that it mapped to a 57 nucleotide sequence, consisting of an AU-rich region containing two AUUUA motifs and a U-rich region containing three UUUUU motifs. An internal deletion showed that the U-rich region was required for inhibition. Point mutations in the AUUUA- and UUUUU-motifs inactivated the inhibitory sequence. Analysis of the stability of mRNAs containing the AU-rich sequence in sense or anti-sense orientation showed that mRNAs containing the AU-rich sequence in sense orientation had reduced half life. Analysis of RNA-protein interactions revealed that binding to the inhibitory sequence of three poly(U) binding proteins (38, 44 and 46 kDa) correlated with inhibition and that the same proteins bind to the AU rich mRNA instability element in the 3' untranslated region on the human c-fos mRNA. We speculate that the human papillomavirus late mRNAs, produced from several hundred copies of the virus genome present in infected cells, compete with the c-fos mRNAs for destabilizing cellular factors and that this may lead to elevated Fos protein levels in human papillomavirus infected cells. PMID- 9393876 TI - Effects of the inhibition of p38/RK MAP kinase on induction of five fos and jun genes by diverse stimuli. AB - The ERK, JNK/SAPK and p38/RK MAP kinase subtypes are differentially activated by physiological, pharmacological and stress stimuli; all three subtypes are implicated in immediate-early (IE) gene induction by these agents. Here, we have asked whether inhibition of a single MAP kinase subtype under these conditions would generally alter induction of several IE genes in a similar way or whether this would differentially up- and down-regulate particular IE genes, an issue which bears on the question of whether individual MAP kinases are strictly targeted to specific IE genes, or whether they might catalyse phosphorylation events that affect several IE genes in the same way. SB 203580, an inhibitor of p38/RK, has been used to analyse the role of this kinase in the induction of five IE genes (c-fos, fosB, c-jun, junB and junD) under diverse conditions of stimulation. In C3H 10T1/2 cells, p38/RK and its downstream kinase MAPKAP K-2 are activated by all stimuli used with the exception of TPA. The specificity of SB 203580 as a p38/RK inhibitor in these cells is demonstrated; it does not affect ERKs or JNK/SAPKs but does result in a small increase in the activity of the upstream kinase MKK6, the principal p38/RK activator in these cells. We find that inhibition of p38/RK under these conditions produces general effects on all five IE genes as a group in three ways. First, induction of all five genes in response to okadaic acid or tumour necrosis factor-alpha (TNF-alpha) is not significantly altered by SB 203580. Second, in cells stimulated with anisomycin or U.V. radiation, SB 203580 potently inhibits all of the induced IE genes. Finally, SB 203580 enhances induction of all five IE genes in EGF-treated cells; these enhanced mRNA levels are not due to stabilisation of labile mRNA transcripts. The significance of these results to current thinking on the relationship between distinct MAP kinase subtypes and specific IE genes is discussed. PMID- 9393877 TI - The Bcr-Abl tyrosine kinase activates mitogenic signaling pathways and stimulates G1-to-S phase transition in hematopoietic cells. AB - Bcr-Abl is a constitutively active tyrosine kinase that is expressed in Philadelphia chromosome (Ph1)-positive human leukemias. Bcr-Abl has been shown to inhibit apoptosis and cause anchorage independent growth. However, its ability to activate mitogenic signaling pathways is controversial. Here we show that Bcr-Abl signaling prevents down-regulation of cyclin-dependent kinase activity and cell cycle arrest after growth factor deprivation of hematopoietic progenitor cells. Using an inducible system to regulate Bcr-Abl expression, we also demonstrate that Bcr-Abl expression is sufficient to induce G1-to-S phase transition, DNA synthesis, and activation of cyclin-dependent kinases in cells that were arrested in G0 by growth factor deprivation. Furthermore, Bcr-Abl activates Ras, Erk, and Jnk pathways as a primary consequence of expression. These data show that Bcr-Abl is one of a select group of oncogenes that is capable of both inhibiting apoptosis and deregulating cell proliferation. The combination of these activities is likely to be important for the progression of CML. PMID- 9393878 TI - Regulation of DNA-dependent protein kinase activity in leukemic cells. AB - The DNA-dependent protein kinase (DNA-PK) complex is composed of a catalytic (DNA PKcs), and a regulatory subunit (Ku70/Ku86 heterodimer). The expression and function of DNA-PK subunits was investigated in purified blood lymphocytes obtained from patients with chronic lymphocytic leukemia (CLL) either refractory to chemotherapy or untreated. Variations in DNA-PK activity were found amongst CLL samples by comparison to human cell lines. It was noticeable that the low DNA PK activity was associated with samples from untreated patients that exhibited a sensitivity phenotype, determined in vitro, to the radiomimetic agent neocarcinostatin by comparison to samples from refractory patients. The regulation in DNA-PK activity was associated with Ku heterodimer expression while DNA-PKcs was unaffected. Moreover, the presence of an altered form of the Ku86 subunit was identified in samples with low DNA-PK activity. These results suggest a regulation process of the DNA-PK activity in fresh human cells. PMID- 9393879 TI - Retinoic acid enhances the expression of interferon-induced proteins: evidence for multiple mechanisms of action. AB - Retinoic acid (RA) and interferons (IFNs) are negative regulators of cell proliferation. In vitro and in vivo, their combination leads to a more potent growth inhibition. However, the molecular mechanisms by which RA and IFNs potentiate each other are not fully understood. As some IFN-induced gene products regulate cell growth and/or antiviral activity, we analysed the effects of RA on their expressions. RA increases the level of 2'5'oligoadenylate synthetase, p68 kinase, the promyelocytic leukemia protein (PML) and Sp100 in both HL-60 and WISH cells. Moreover, RA and IFN act cooperatively to increase the expression of these proteins. RA also inhibits vesicular stomatitis virus replication and induces a higher antiviral state and growth inhibition when combined with IFN. RA stimulates the IFN regulatory factor 1 (IRF-1) gene expression directly through the GAS motif and causes the induction and secretion of IFNalpha. Additional mechanisms could be involved as RA increases the level of signal transducing activators of transcription (STAT) proteins, and enhances the IFN-induced STAT activation, suggesting that cooperative effects by RA and IFN are mediated through multiple pathways. PMID- 9393880 TI - Praja1, a novel gene encoding a RING-H2 motif in mouse development. AB - As part of a cloning strategy to identify genes involved in early mouse liver development we have isolated Praja1, a gene with similar sequences to the Drosophila melanogaster gene goliath (gl) which is involved in the fate of mesodermal cells ultimately forming gut musculatures, fat body, and the heart. Praja1 is a 2.1 kb gene encoding a putative 396 amino acid ORF and includes a COOH-terminal RING-H2 domain. Using the Jackson Laboratory BSS panel, we have localized Praja1 on chromosome X at 36 cM, which may be a candidate gene for mouse sla (sex linked sideroblastic anemia), near the X inactivation center gene, Xist. Northern blot analysis demonstrated three transcripts (3.1, 2.6 and 2.1 kb) in mRNA from adult mouse tissues brain, liver, and kidney as well as in mRNA from developing mouse embryos (days 7, 11, 15 and 17 post coitus, p.c.). In vitro transcription/translation yielded a product with an Mr of 59 kD. Immunohistochemical staining of in vitro liver explant cultures using a heterologous antibody against praja1 demonstrated cytoplasmic staining of cuboidal cells that have hepatocyte morphology and organization. The presence of the RING-H2 domain, a proline-rich region at the COOH-end, and regions rich in acidic amino acids, leads to the hypothesis that the Praja1 product is possibly involved in mediating protein-protein interactions, possibly as part of a protein sorting or transport pathway. This is strengthened by the similarity of Praja1 to rat Neurodap1, whose product has been shown to localize to the endoplasmic reticulum and golgi in brain. PMID- 9393881 TI - Deletional remodeling of c-myc-deregulating chromosomal translocations. AB - Evidence is presented for the existence of a novel remodeling-by-deletion mechanism that alters the fine structure of c-myc-deregulating chromosomal translocations in t(12;15)-positive BALB/c plasmacytomas. DNA sequence analysis of the t(12;15) in five primary tumors revealed the co-existence of precursor cells harboring genetic recombinations between the immunoglobulin heavy-chain mu locus (Igh mu) and c-myc with clonally related progenitors containing rearrangements between the immunoglobulin heavy-chain alpha locus (Igh alpha) and c-myc. Clonal relatedness was based upon unique junction fragments between the switch region of Igh mu and c-myc. S mu/c-myc junctions are thus useful clonotypic markers for monitoring the conversion of Igh mu/c-myc-positive tumor precursor clones into Igh alpha/c-myc-positive plasmacytomas. Aberrant isotype switch recombination appears to be the most likely mechanism effecting this conversion event (other possibilities are discussed) which may help to explain the preferred usage of the Igh alpha locus in recombinations with c-myc in t(12;15)-positive plasma cell tumors in BALB/c mice. PMID- 9393883 TI - Neuroprotection. PMID- 9393882 TI - The phosphatidylinositol polyphosphate 5-phosphatase SHIP and the protein tyrosine phosphatase SHP-2 form a complex in hematopoietic cells which can be regulated by BCR/ABL and growth factors. AB - We report here that interleukin-3 (IL-3) and erythropoietin (EPO) induce formation of a complex composed of two SH2-containing phosphatases, the tyrosine phosphatase SHP-2 and the SH2 containing inositol 5-phosphatase (SHIP). Both SHP 2 and SHIP are known to be involved in growth factor signal transduction, but their potential interaction in the same pathway is novel. SHIP has previously been shown to associate with SHC, and potentially to be involved in regulating apoptosis. In contrast, in some model systems, SHP-2 has been demonstrated to positively regulate cell growth. Both phosphatases in the complex were tyrosine phosphorylated, and the amount of SHIP coprecipitating with SHP-2 was inversely related to the amount of SHIP coprecipitating with SHC. In hematopoietic cells transformed by the BCR/ABL oncogene, this phosphatase complex was found to be constitutively present with both components heavily tyrosine phosphorylated. Also, other proteins were detected in the complex, including BCR/ABL itself and c CBL. However, transformation by BCR/ABL was associated with a reduced SHIP protein expression, which could further affect the accumulation of various inositol polyphosphates in these leukemic cells. These data suggest that the function of SHIP and SHP-2 in normal cells are linked and that BCR/ABL alters the function of this signaling complex. PMID- 9393885 TI - Ischaemic cerebral injury, intrauterine growth retardation, and placental infarction. AB - Two hundred and twenty-five consecutive autopsies performed on fetuses >20 weeks' gestation were reviewed, and 37 growth-retarded stillborn fetuses without multiple congenital abnormalities or evidence of intrauterine infection were identified. Histological evidence of ischaemic cerebral injury was found in 31 of the 37 cases and placental infarction was seen in 26 of 36 placentas. Of the 31 cases with cerebral ischaemia, 24 had placental infarcts. Twenty-six of 27 stillborn fetuses >26 weeks' gestation showed histological evidence of cerebral ischaemia. It was concluded that in the group of growth-retarded fetuses studied, there was a high incidence of both cerebral and placental ischaemic abnormality. PMID- 9393884 TI - Relation of deranged neonatal cerebral oxidative metabolism with neurodevelopmental outcome and head circumference at 4 years. AB - Cerebral oxidative metabolism was studied using phosphorus magnetic resonance spectroscopy during the first week of life and neurodevelopmental outcome was assessed at 4 years in 62 infants who had clinical and/or biochemical evidence consistent with birth asphyxia (critically impaired intrapartum gas exchange). Twenty-one died and the neurodevelopmental status of the 41 who survived was assessed by a range of tests at age 4 years. The minimum recorded values for the cerebral phosphocreatine:inorganic phosphate concentration ratio (an index of oxidative metabolism) were related to outcome. The results showed significant relations between the extent of derangement of neonatal oxidative metabolism and a range of adverse outcomes, including death, and at 4 years reduced head growth and the presence and severity of neuromotor impairments, overall neurodevelopmental impairments, and cognitive functioning. Strong correlations between the extent of derangement of neonatal oxidative metabolism and outcome at 1 and 4 years were also shown. We conclude that the severities of adverse outcomes at 1 and 4 years of age were closely related to the extent of cerebral energy derangement in the first week of life, and we also conclude that primary intrapartum hypoxic-ischaemic cerebral injury was generally responsible for the events that led to death, microcephaly, and impaired PMID- 9393886 TI - The effect of seizure type and medication on cognitive and behavioral functioning in children with idiopathic epilepsy. AB - Antiepileptic drugs have been reported to have a variety of adverse effects on behavior and performance in children with epilepsy. Previous studies investigating these side effects, however, have not controlled for the baseline status of the child (e.g. underlying neurological condition, seizure type, socioeconomic status, family variables), making it difficult to determine whether changes in function are attributable to the use of medication. We investigated the cognitive and behavioral profiles of 43 children, aged from 4 to 16 years, with new onset, idiopathic seizures. Twenty-six of these children participated in a 6-month follow-up study, and 12 in a 12-month follow-up study, investigating the effects of antiepileptic medications on psychological functioning. The children were of average intelligence (mean IQ 108) and had not previously been treated with antiepileptic medication. Children were classified as having either generalized convulsive, generalized non-convulsive (absence), simple partial, or complex partial seizures. Prior to the initiation of treatment, children with partial seizures were found to perform better than children with generalized seizures on measures of cognitive functioning. Children with convulsive seizures obtained significantly higher cognitive scores than those with non-convulsive seizures. Children with generalized non-convulsive seizures had lower cognitive scores than subjects with other types of seizure. No differences were found between groups at baseline prior to the initiation of antiepileptic medications. Analysis of subjects' performance after 6 and 12 months of antiepileptic therapy showed no significant deterioration attributable to medication. The differences in cognitive performance of the four seizure groups at baseline were not apparent at the time of follow-up. These results indicate that intrinsic and environmental variables may play a more significant role in predisposing certain children to cognitive and learning problems than do antiepileptic medications. PMID- 9393887 TI - Covert orienting of visuospatial attention in children with developmental coordination disorder. AB - It is still unclear whether impairments in visuospatial processing in children with developmental coordination disorder (DCD) are a consequence of their motor deficits or are independent of them. In two experiments, 20 children with DCD and 20 matched controls were tested on the covert orienting of a visuospatial attention task (COVAT). Experiment 1 used a COVAT with peripheral cues and an 80% probability that targets would appear at the cued location. While the results suggested a deficit in the disengage operation of orienting covert attention for the DCD group, they were difficult to reconcile with models of covert orienting and the results of past research. Experiment 2 tested subjects on two new versions of the COVAT: the first used peripheral cues and no probability information (exogenous mode), and the second used central cues and an 80% probability that targets would appear at the cued location (endogenous mode). The DCD group displayed attentional orienting deficits only for the endogenous mode. These results suggest that impairments in the endogenous control of visuospatial attention are independent of motor deficits in DCD. PMID- 9393888 TI - Caregivers' perceptions following gastrostomy in severely disabled children with feeding problems. AB - Feeding difficulties are common in neurologically impaired children, often leading to great distress and frustration in the child and family. A gastrostomy may be advocated if oral intake is inadequate causing poor weight gain or when there is significant aspiration during feeding, or if feeding is very distressing. To find out if caregivers were happy with the outcome of gastrostomy (with fundoplication, when indicated), a 35-item questionnaire was developed and sent to 38 of them. Twenty-nine replies were received and appeared to be representative of the whole group. Coughing, choking, and vomiting improved in most cases. Weight gain improved in all in whom it had been a problem. In the majority, it became easier to give the children their medications although control of epilepsy was unchanged overall. Time spent feeding the child was reduced and many caregivers had more time to devote to other children and themselves. Only one parent regretted the operation. In children with severe disability and feeding problems, a gastrostomy (with fundoplication if there is significant reflux) can reduce symptoms of vomiting, coughing, and choking, help growth and improve quality of life in the child, when patients are properly selected. PMID- 9393889 TI - Correlates of maladaptive behavior in individuals with 5p- (cri du chat) syndrome. AB - This study examined the range, distinctiveness, and correlates of maladaptive behavior in 146 subjects with 5p- (cri du chat) syndrome using the Aberrant Behavior Checklist as a standardized measure. Hyperactivity was the most significant and frequent problem in the sample. Subjects with 5p- syndrome also showed aggression, tantrums, self-injurious behavior, and stereotypies; some of these problems were more pronounced in individuals with lower cognitive-adaptive levels, as well as in those with histories of previous medication trials. Autistic-like features and social withdrawal were more characteristic of individuals with translocations as opposed to deletions, even when controlling for the lower adaptive level of the translocation group. These findings encourage further research on the behavior of individuals with 5p- syndrome. PMID- 9393890 TI - Families of children with 5p- (cri du chat) syndrome: familial stress and sibling reactions. AB - This research examined family stress and sibling reactions in families of children with 5p- (cri du chat) syndrome aged 1 to 18 years who were living at home. In Study 1, 99 parents reported on themselves and their child with 5p-, as well as on family demographics, social supports, and stress. The best predictor of familial stress was the child's amount of maladaptive behavior, accounting for 12 to 38% of the variance across different stress measures. In Study 2, sibling concerns were examined in 44 unaffected siblings. The major finding was that parents and siblings disagreed on the extent of the siblings' interpersonal concerns. Parents reported that siblings felt ignored and misunderstood, whereas siblings themselves rated these concerns at much lower levels. PMID- 9393891 TI - Neurobrucellosis in childhood: six new cases and a review of the literature. AB - Neurobrucellosis accounts for <1% of cases of brucellosis in children. Six new cases of neurobrucellosis are presented and data from 39 previously published cases are analysed. The incidence is equal in males and females, and the source of infection is likely to be unpasteurised milk. Clinical presentation varies from severe meningoencephalitis or peripheral neuropathy/radiculopathy to behavioural disturbance. Diagnostic certainty requires isolation of the organism from the CSF, but as this is rarely possible serological diagnosis can be performed with the Coombs test on the CSF. Treatment requires combination antibiotic therapy and should continue for at least 8 weeks. PMID- 9393892 TI - Fast-wave periodic complexes in a mentally retarded child who later developed subacute sclerosing panencephalitis: a modification of a classic EEG by preexisting brain damage? AB - The EEG of a 12-year-old girl with stage II subacute sclerosing panencephalitis (SSPE), who had also suffered from a non-progressive mental retardation of unknown aetiology since early childhood, revealed periodic generalised stereotyped fast wave bursts synchronous with myoclonic jerks. The background activity was nearly normal. The diagnosis of SSPE was established by raised serum and measles antibody titres, raised CSF IgG, and brain biopsy. This rare type of periodic complex has only once been described in the literature, again in a mentally retarded child who had developed SSPE. We suggest a mechanism of origin of this type of periodic complex drawn from observations in these two cases, and discuss its significance. PMID- 9393893 TI - Absence of alpha-sarcoglycan and novel missense mutations in the alpha sarcoglycan gene in a young British girl with muscular dystrophy. AB - An 11-year-old white female presented with progressive proximal muscle weakness and marked calf hypertrophy. Muscle biopsy showed severe dystrophy with normal expression of dystrophin. There was complete absence of the 50kDa dystrophin associated glycoprotein (alpha-sarcoglycan). DNA analysis showed novel point mutations (one missense and one splicing) in the alpha-sarcoglycan gene at chromosomal location 17q21, confirming the diagnosis of limb-girdle muscular dystrophy type 2D (LGMD-2D). We believe this is one of the first confirmed white cases of primary alpha-sarcoglycanopathy identified in the UK. This case supports the assumption of a wide geographic prevalence of severe childhood onset autosomal recessive muscular dystrophy and genetic heterogeneity. In the future, with improved diagnostic accuracy it is likely that more cases demonstrating primary or secondary deficiency of alpha-sarcoglycan will be identified. We would recommend staining for dystrophin-associated glycoproteins (sarcoglycans) in all new cases of muscular dystrophy with normal dystrophin, and confirmation with DNA analysis where possible. PMID- 9393895 TI - Epilepsy and prejudice with particular relevance to childhood. PMID- 9393894 TI - Myositis ossificans complicating severe Guillain-Barre syndrome. AB - We report myositis ossificans occurring in a 13-year-old boy with severe and rapidly progressive Guillain-Barre syndrome. This complication should be considered when severe musculoskeletal pain is experienced by such patients. Disodium etidronate may be of benefit in this condition. PMID- 9393896 TI - The Worster-Drought syndrome: a severe test of paediatric neurodisability services? PMID- 9393897 TI - Thromboprophylaxis in elective orthopaedic surgery -- what is the purpose? PMID- 9393898 TI - Fracture of the femur in children. PMID- 9393899 TI - Planovalgus and cavovarus deformity of the hind foot. A functional approach to management. PMID- 9393901 TI - Total hip arthroplasty with an uncemented femoral component. Excellent results at ten-year follow-up. AB - We followed 138 patients (145 hips) who had had uncemented total hip arthroplasty using the Taperloc femoral component for a mean of ten years (8 to 12.5). No patient was lost to follow-up; 31 (31 hips) died before the minimum time of eight years for inclusion in the study, and 30 of these still had their femoral component in place. One well-fixed prosthesis had been exchanged at the time of acetabular revision. Of the remaining 114 hips, one femoral component required revision for aseptic loosening and one for sepsis. Three other well-fixed femoral components were removed during acetabular revision. Complete clinical and radiological follow-up was obtained in the 109 hips which had not had revision. Clinically, 94 (87%) were rated good or excellent, eight (7%) fair and seven (6%) poor. The average Harris hip score increased from 48 before operation to 88 at the time of the last follow-up. Radiologically, 103 hips (94%) had fixation by bone ingrowth, three (3%) showed stable fibrous ingrowth and three (3%) were unstable. Osteolysis of the fomoral cortex was seen in seven hips (6%), with major lysis in only one. At a mean follow-up of ten years, the results of the Taperloc femoral component are comparable with those of modern techniques of cementing in primary total hip arthroplasty. PMID- 9393900 TI - Mortality and fatal pulmonary embolism after primary total hip replacement. Results from a regional hip register. AB - We calculated the rates for perioperative mortality and fatal pulmonary embolism (PE) after primary total hip replacement in a single UK health region, using a regional arthroplasty register and the tracing service of the Office of National Statistics. During 1990, there were 2111 consecutive primary replacements in 2090 separate procedures. Within 42 days of operation a total of 19 patients had died (0.91%, 95% CI 0.55 to 1.42). Postmortem examination showed that four deaths (0.19%, 95% CI 0.05 to 0.49) were definitely due to PE. The overall perioperative mortality and fatal PE rates are low and in our study did not appear to be altered by the use of chemical thromboprophylaxis (perioperative mortality rate: one-tailed Fisher's exact test, p = 0.39; fatal PE rate: one-tailed Fisher's exact test, p = 0.56). The routine use of chemical thromboprophylaxis for primary THR is still controversial. The issue should be addressed by an appropriate randomised, prospective study using overall mortality and fatal PE rate as the main outcome measures, but the feasibility of such a study is questioned. PMID- 9393902 TI - The cement mantle in femoral impaction allografting. A comparison of three systems from four centres. AB - An analysis of the cement mantle obtained with the Exeter impaction allografting system at one centre showed that it was either deficient or absent in almost 47% of Gruen zones. We therefore examined the mantle obtained using this system at another hospital and compared the results with those from the CPT and Harris Precoat Systems at other centres. The surgical indications for the procedure and the patient details were broadly similar in all four hospitals. There was some variation in the frequency of use of cortical strut allografts, cerclage wires and wire mesh to supplement the impaction allograft. Analysis of the cement mantles showed that when uncertain Gruen zones were excluded, the incidence of zones with areas of absence or deficiency of the cement was 47% and 50%, respectively, for the two centres using the Exeter system, 21% for the CPT system and 18% for the Harris Precoat system. We measured the difference in size between the proximal allograft impactors and the definitive prosthesis for each system. The Exeter system impactors are shorter than the definitive prosthesis and taper sharply so that the cavity created is inadequate, especially distally. The CPT proximal impactors are considerably longer than the definitive prosthesis and are designed to give a mantle of approximately 2 mm medially and laterally and 1.5 mm anteriorly and posteriorly. The Harris Precoat proximal impactors allow for a mantle with a circumference of 0.75 mm in the smaller sizes and 1 mm in the larger. Many reports link the longevity of a cemented implant to the adequacy of the cement mantle. For this reason, femoral impaction systems require careful design to achieve a cement mantle which is uninterrupted in its length and adequate in its thickness. Our results suggest that some current systems require modification. PMID- 9393903 TI - Continuous passive motion after primary total knee arthroplasty. Does it offer any benefits? AB - We report a prospective randomly controlled trial to examine the effectiveness of continuous passive motion (CPM) in improving postoperative function and range of movement after total knee arthroplasty (TKA). We allocated 53 patients (57 knees) to one of three postoperative regimes: no CPM (n = 19); CPM at 0 to 40 degrees (0 to 40 CPM; n = 18); and CPM at 0 to 70 degrees (0 to 70 CPM; n = 20). Those in the CPM groups had CPM for 48 hours and all patients had an identical regime of physiotherapy. There was an even distribution of various cemented and cementless TKAs in each group. Patients were assessed preoperatively and at one week and at one year postoperatively. At one week, there was a statistically significant increase in the range of flexion and total range of movement in the 0 to 70 CPM group compared with the no-CPM group. At one year we found no significant differences in mean flexion, overall range of movement, fixed flexion deformity or functional results in the three groups. Those who had CPM had a significant increase in analgesic requirement (p = 0.04). There was an increased mean blood drainage postoperatively in those who had 0 to 70 CPM (1558 ml) compared with those with no CPM (956 ml) (t = 2.96, p = 0.005) and with 0 to 40 CPM (1017 ml) (t = 2.62, p = 0.01). Our findings show that CPM had no significant advantage in terms of improving function or range of movement, and that its use increased blood loss and analgesic requirements. PMID- 9393904 TI - Synovectomy of the elbow and radial head excision in rheumatoid arthritis. Predictive factors and long-term outcome. AB - We carried out a survival analysis of elbow synovectomy (ES) and excision of the radial head (RHE) performed on 171 rheumatoid elbows. The failure criteria were revision surgery (performed or desired) and/or the presence of significant or severe pain. The cumulative survival was 81% at one year which thereafter decreased by an average of 2.6% per year. The strongest predictor for success was a low preoperative range of supination-pronation when corresponding survival curves were compared. A low range of flexion-extension also predicted failure. Combining both factors gave better prediction (failure: 6.3% v 67%), but a long duration of elbow symptoms before surgery predicted failure (72%, p = 0.04). At review, there was a mean gain of 50 degrees in supination-pronation and 11 degrees in flexion-extension; both correlated with success. Failure correlated with recurrence of synovitis, elbow instability, ulnar neuropathy, poor general mobility and poor upper-limb function. The last was independently affected by the severity of RA in the ipsilateral shoulder. Our findings show that although the short-term result of ES and RHE in rheumatoid arthritis is good, the long-term outcome is poor except in a subgroup with more than 50% limitation of forearm rotation. PMID- 9393905 TI - Forequarter amputation for high-grade malignant tumours of the shoulder girdle. AB - We reviewed 20 patients after forequarter amputation performed for high-grade malignant tumours of the shoulder girdle (Enneking grades IIB to III). The operations were classified as palliative or curative according to the resection margins and the presence of disseminated disease at the time of the surgery. There were five palliative and 15 curative procedures. Two patients died from unrelated causes, septicaemia and suicide. Eight died in the first two years, four of whom had had a palliative operation. Four died between two and five years after surgery, one after a palliative operation. Five patients are alive, at a mean of 89.4 months after surgery, four of whom are free from disease. The median survival after a palliative amputation was 20.6 months. Our overall five-year survival (palliative and curative cases) was 21.2%, for curative cases it was 30.2%. None of the patients use an artificial prosthesis. Despite the disfigurement which results from this operation, it still has a useful role to play in the management of high-grade malignant tumours of the upper limb. PMID- 9393906 TI - Stanmore custom-made extendible distal femoral replacements. Clinical experience in children with primary malignant bone tumours. AB - The use of extendible distal femoral replacements is a relatively new treatment alternative for malignant bone tumours in growing individuals. Although their appearance was widely appreciated, questions about functional practicality and longevity remain unclear. With longer follow-up, advantages of immediate functional restoration and beneficial psychological aspects seem to be overshadowed by an increase in complications such as aseptic loosening, infection or prosthetic failure. We have reviewed 18 children with such tumours who were treated between 1983 and 1990 by custom-made Stanmore extendible distal femoral replacements. Four died from metastatic disease within 2.5 years of operation and two required amputation for local recurrence or chronic infection. The remaining 12 patients were followed for a mean of 8.7 years (6 to 13.2). A mean total lengthening of 5.2 cm was achieved, requiring, on average, 4.3 operations. Using the Musculoskeletal Tumor Society rating score the functional result at review was, on average, 77% of the expected normal function, with seven patients achieving > or = 80%. Revision of the prosthesis was required in ten patients, in six for aseptic loosening, at a mean of 6.2 years after the initial procedure. PMID- 9393907 TI - Growth after extendible endoprosthetic replacement of the distal femur. AB - We report our results in 24 children with malignant primary bone tumours of the distal femur treated with a Stanmore extendible endoprosthesis (SEER). This consists of a femoral component that can be lengthened, a constrained knee and an uncemented sliding tibial component which crosses the proximal tibial physeal plate perpendicularly. The average age of the patients at diagnosis was ten years and the mean follow-up was 4.7 years (2.5 to 7.9). The mean growth of the affected tibia was 76% (18 to 136) and of the fibula 83% (15 to 750) of the growth of the unaffected limb. Measurement of growth arrest lines showed that the mean growth of the proximal tibial physis on the affected side was 69% (43 to 100) of that of the normal side. The great variability in the growth of the physis cannot yet be explained. PMID- 9393908 TI - Far lateral lumbar disc herniation. The key to the intertransverse approach. AB - Of a total of 330 patients requiring operation on a lumbar disc, 20 (6.1%) with lateral disc prolapse had a new muscle-splitting, intertransverse approach which requires minimal resection of bone. There were 16 men and 4 women with a mean age of 52 years. All had intense radicular pain, 15 had femoral radiculopathy and 19 a neurological deficit. Far lateral herniation of the disc had been confirmed by MRI. At operation, excellent access was obtained to the spinal nerve, dorsal root ganglion and the disc prolapse. The posterior primary ramus was useful in locating the spinal nerve and dorsal root ganglion during dissection of the intertransverse space. At review from six months to four years, 12 patients had excellent results with no residual pain and six had good results with mild discomfort and no functional impairment. Two had poor results. There had been neurological improvement in 17 of the 20 patients. We report a cadaver study of the anatomy of the posterior primary ramus. It is readily identifiable through this approach and can be traced down to the spinal nerve in the intertransverse space. We recommend the use of a muscle-splitting intertransverse approach to far lateral herniation of the disc, using the posterior primary ramus as the key to safe dissection. PMID- 9393909 TI - Neurological deterioration after posterior wiring of the cervical spine. AB - Posterior cervical wiring is commonly performed for patients with spinal instability, but has inherent risks. We report eight patients who had neurological deterioration after sublaminar or spinous process wiring of the cervical spine; four had complete injuries of the spinal cord, one had residual leg spasticity and three recovered after transient injuries. We found no relation between the degree of spinal canal encroachment and the severity of the spinal cord injury, but in all cases neurological worsening appeared to have been caused by either sublaminar wiring or spinous process wiring which had been placed too far anteriorly. Sublaminar wiring has substantial risks and should be used only at atlantoaxial level, and then only after adequate reduction. Fluoroscopic guidance should be used when placing spinous process wires especially when the posterior spinal anatomy is abnormal. PMID- 9393910 TI - Sparing of sensation to pin prick predicts recovery of a motor segment after injury to the spinal cord. AB - We have reviewed 59 patients with injury to the spinal cord to assess the predictive value of the sparing of sensation to pin prick in determining motor recovery in segments which initially had MRC grade-0 power. There were 35 tetraplegics (18 complete, 17 incomplete) and 24 paraplegics (19 complete, 5 incomplete), and the mean follow-up was 29.6 months. A total of 114 motor segments initially had grade-0 power but sparing of sensation to pin prick in the corresponding dermatome. Of these, 97 (85%) had return of functional power (> or = grade 3) at follow-up. There were 479 motor segments with grade-0 power but no sparing of sensation to pin prick and of these only six (1.3%) had return of functional power. Both of the above associations were statistically significant (chi-squared test, p < 0.0001). After injury to the spinal cord, the preservation of sensation to pin prick in a motor segment with grade-0 power indicated an 85% chance of motor recovery to at least grade 3. PMID- 9393911 TI - Lengthening of short great toes by callus distraction. AB - We lengthened seven first metatarsals in four patients with short great toes by callus distraction using an external fixator. Good clinical and cosmetic results were obtained. Bone lengthening is effective in patients with short great toes not only for cosmesis, but also to relieve pain and callosities on the plantar aspect of the second and third metatarsal heads. Excessive lengthening of the first metatarsal resulted in limitation of the range of movement of the metatarsophalangeal joint of the great toe. To prevent this the amount of lengthening should not exceed 40% of the preoperative length of the metatarsal. PMID- 9393913 TI - The fibula-flexor hallucis longus osteomuscular flap. AB - Limb salvage after loss of bone and soft tissue may require many operations to obtain soft-tissue cover and bony continuity. We describe a fibula-flexor hallucis longus osteomuscular flap which can provide both soft tissue and bone in a single stage. The flap is based on the peroneal vessels and is covered by a split-thickness skin graft. We report the results in five patients with an average bone defect of 8.3 cm and soft-tissue and skin loss. All regained a normal gait on the donor side; four had clinical and radiological union with excellent soft-tissue cover, but one required later amputation due to diffuse coagulopathy. The flap provides free vascularised bone with muscle cover. It has a dependable, long pedicle with minimal morbidity at the donor site, and allows monitoring of the vascularity of the fibular graft. PMID- 9393912 TI - Does indomethacin reduce heterotopic bone formation after operations for acetabular fractures? A prospective randomised study. AB - We have studied prospectively the effect of indomethacin on the development of heterotopic ossification (HO) after the internal fixation of acetabular fractures. After operation 107 patients randomly received either a six-week course of indomethacin or no treatment against HO. Plain radiographs of 101 patients at a mean of 7.9 months after surgery showed HO in 47.4% of the 57 patients who received indomethacin and in 56.8% of the 44 who did not. This difference was not statistically significant. Heterotopic ossification of Brooker class II or more was seen in four patients (7%) with prophylaxis and in one without (p = 0.51). Measurements of the volume of HO on 3-D CT reconstructions showed a median value of 1.5 cm3 in patients with indomethacin and 4.0 cm3 in those without (p = 0.28). When only the 57 patients in whom the operation was carried out through either a Kocher-Langenbeck or an extended iliofemoral approach were included the indomethacin group showed a median volume of 1.7 cm3 compared with 3.6 cm3. On plain radiographs Brooker class II or above was seen in 9.4% of the patients receiving indomethacin and in 4.8% of those who did not. Indomethacin was therefore not effective in preventing ectopic bone formation after surgery for acetabular fractures. There was a significant association of male gender with volume of HO using a non-parametric analysis of variance. PMID- 9393914 TI - Awareness of tip-apex distance reduces failure of fixation of trochanteric fractures of the hip. AB - We compared the results of the surgical treatment of trochanteric hip fractures before and after surgeons had been introduced to the tip-apex distance (TAD) as a method of evaluating screw position. There were 198 fractures evaluated retrospectively and 118 after instruction. The TAD is the sum of the distance from the tip of the screw to the apex of the femoral head on anteroposterior and lateral views. This decreased from a mean of 25 mm in the control group to 20 mm in the study group (p = 0.0001). The number of mechanical failures by cut-out of the screw from the head decreased from 16 (8%) in the control group at a mean of 13 months to none in the study group at a mean of eight months (p = 0.0015). There were significantly fewer poor reductions in the study group. Our study confirms the importance of good surgical technique in the treatment of trochanteric fractures and supports the concept of the TAD as a clinically useful way of describing the position of the screw. PMID- 9393915 TI - A more accurate method of measurement of angulation after fractures of the tibia. AB - Accurate measurement of the alignment of the tibia is important both clinically and in research. The conventional method of measuring the angle of malunion after a fracture of the shaft of the tibia is potentially inaccurate because the mechanical axis of the normal bone may not pass down the centre of the medullary canal. An alternative method is described in which a radiograph of the opposite tibia is used as a template. A sample of 56 sets of standard radiographs of healed fractures of the shaft of the tibia was evaluated. The 95% limits of agreement between this and the conventional method were wide, being -6.2 degrees to +5.5 degrees for coronal angulation and -6.7 degrees to +8.1 degrees for sagittal angulation. These results suggest that the conventional method is inaccurate. The new method has good inter- and intraobserver reliability. PMID- 9393916 TI - External fixation or flexible intramedullary nailing for femoral shaft fractures in children. A prospective, randomised study. AB - We report the outcome of 19 children aged 5.2 to 13.2 years with 20 fractures of the femoral shaft requiring surgery, who were randomly assigned to have external fixation (EF) or flexible intramedullary nailing (FIN) (10 fractures each). The duration of the operation averaged 56 minutes for the EF group with 1.4 minutes of fluoroscopy, compared with 74 minutes and 2.6 minutes, respectively, for the FIN group. The early postoperative course was similar, but the FIN [corrected] group showed much more callus formation. The time to full weight-bearing, full range of movement and return to school were all shorter in the FIN group. The FIN complications included one transitory foot drop and two cases of bursitis at an insertion site. In the EF group there was one refracture, one rotatory malunion requiring remanipulation and two pin-track infections. At an average follow-up of 14 months two patients in the EF group had mild pain, four had quadriceps wasting, one had leg-length discrepancy of over 1 cm, four had malalignment of over 5 degrees, and one had limited hip rotation. In the FIN group, one patient had mild pain and one had quadriceps wasting; there were no length discrepancies, malalignment or limitation of movement. Parents of the FIN group were more satisfied. We recommend the use of flexible intramedullary nailing for fractures of the femoral shaft which require surgery, and reserve external fixation for open or severely comminuted fractures. PMID- 9393917 TI - Different methods of treatment related to the bilateral occurrence of Perthes' disease. AB - We treated 98 consecutive patients with Perthes' disease by a unilateral brace in external rotation, flexion and abduction and a further consecutive 110 by a bilateral cast with the hips in internal rotation and abduction. During treatment in the unilateral brace, six (6.1%) hips on the opposite side developed evidence of Perthes' disease and one developed this after the brace had been removed. In children managed in bilateral casts, no contralateral Perthes' disease was seen. Adequate containment of the femoral head may prevent subsequent changes in the opposite hip. PMID- 9393918 TI - Primary subacute haematogenous osteomyelitis of the tarsal bones in children. AB - Primary subacute haematogenous osteomyelitis (PSHO) of the small bones of the foot is a rare and infrequently considered cause of a limp in children. We describe 11 patients with PSHO, of whom nine were under three years of age, who had a limp with few symptoms. The talus was involved in 36%. Bone scans were positive in all patients and led to localisation of the lesion in two. The radiological features included soft-tissue swelling, an osteolytic lesion in the talus and the calcaneus and a sclerotic appearance of the cuboid and the navicular bones. All patients except one were cured with antibiotics. PMID- 9393919 TI - Regulation of bone cells by particle-activated mononuclear phagocytes. AB - Bone loss around replacement prostheses may be related to the activation of mononuclear phagocytes (MNP) by prosthetic wear particles. We investigated how osteoblast-like cells were regulated by human MNP stimulated by particles of prosthetic material. Particles of titanium-6-aluminium-4-vanadium (TiAIV) stimulated MNP to release interleukin (IL)-1beta, tumour necrosis factor (TNF)alpha, IL-6 and prostaglandin E2 (PGE2). All these mediators are implicated in regulating bone metabolism. Particle-activated MNP inhibited bone cell proliferation and stimulated release of IL-6 and PGE2. The number of cells expressing alkaline phosphatase, a marker associated with mature osteoblastic cells, was reduced. Experiments with blocking antibodies showed that TNFalpha was responsible for the reduction in proliferation and the numbers of cells expressing alkaline phosphatase. By contrast, IL-1beta stimulated cell proliferation and differentiation. Both IL-1beta and TNFalpha stimulated IL-6 and PGE2 release from the osteoblast-like cells. Our results suggest that, particle activated mononuclear phagocytes can induce a change in the balance between bone formation and resorption by a number of mechanisms. PMID- 9393920 TI - Age variations in the properties of human tibial trabecular bone. AB - We tested in compression specimens of human proximal tibial trabecular bone from 31 normal donors aged from 16 to 83 years and determined the mechanical properties, density and mineral and collagen content. Young's modulus and ultimate stress were highest between 40 and 50 years, whereas ultimate strain and failure energy showed maxima at younger ages. These age-related variations (except for failure energy) were non-linear. Tissue density and mineral concentration were constant throughout life, whereas apparent density (the amount of bone) varied with ultimate stress. Collagen density (the amount of collagen) varied with failure energy. Collagen concentration was maximal at younger ages but varied little with age. Our results suggest that the decrease in mechanical properties of trabecular bone such as Young's modulus and ultimate stress is mainly a consequence of the loss of trabecular bone substance, rather than a decrease in the quality of the substance itself. Linear regression analysis showed that collagen density was consistently the single best predictor of failure energy, and collagen concentration was the only predictor of ultimate strain. PMID- 9393921 TI - Synthetic osteochondral replacement of the femoral articular surface. AB - We have studied damage to the tibial articular surface after replacement of the femoral surface in dogs. We inserted pairs of implants made of alumina, titanium and polyvinyl alcohol (PVA) hydrogel on titanium fibre mesh into the femoral condyles. The two hard materials caused marked pathological changes in the articular cartilage and menisci, but the hydrogel composite replacement caused minimal damage. The composite osteochondral device became rapidly attached to host bone by ingrowth into the supporting mesh. We discuss the clinical implications of the possible use of this material in articular resurfacing and joint replacement. PMID- 9393922 TI - Fresh osteochondral allografts for post-traumatic osteochondral defects of the knee. AB - We used fresh small-fragment osteochondral allografts to reconstruct post traumatic osteochondral defects in 126 knees of 123 patients with a mean age of 35 years. At a mean follow-up of 7.5 years (2 to 20), 108 knees were rated as successful (85%) and 18 had failed (15%). The factors related to failure included age over 50 years (p = 0.008), bipolar defects (p < 0.05), malaligned knees with overstressing of the grafts, and workers' compensation cases (p < 0.04). Collapse of the graft by more than 3 mm and of the joint space of more than 50% were seen more frequently in radiographs of failed grafts. Our encouraging clinical results for fresh small-fragment osteochondral allografts show that they are indicated for unipolar post-traumatic osteochondral defects of the knee in young active patients. PMID- 9393923 TI - Lower-limb lengthening in short stature. An electrophysiological and clinical assessment of peripheral nerve function. AB - We assessed peripheral nerve function during and after lower-limb lengthening by callotasis in 14 patients with short stature, using motor conduction studies. Four patients with short stature of varying aetiology showed unilateral and one showed bilateral weakness of foot dorsiflexion. Both clinical and electrophysiological abnormalities consistent with involvement of the peroneal nerve were observed early after starting tibial callotasis. There was some progressive electrophysiological improvement despite continued bone distraction, but two patients with Turner's syndrome had incomplete recovery. A greater percentage increase in tibial length did not correspond to a higher rate of peroneal nerve palsy. The function of the posterior leg muscles and the conduction velocity of the posterior tibial nerve were normal throughout the monitoring period. The F-wave response showed a longer latency at the end of the bone distraction than in basal conditions; this is probably related to the slowing of conduction throughout the entire length of the nerve. PMID- 9393924 TI - Length and torsion of the lower limb. AB - Corrective osteotomies are often planned and performed on the basis of normal anatomical proportions. We have evaluated the length and torsion of the segments of the lower limb in normal individuals, to analyse the differences between left and right sides, and to provide tolerance figures for both length and torsion. We used CT on 355 adult patients and measured length and torsion by the Ulm method. We excluded all patients with evidence of trauma, infection, tumour or any congenital disorder. The mean length of 511 femora was 46.3 +/- 6.4 cm (+/-2SD) and of 513 tibiae 36.9 +/- 5.6 cm; the mean total length of 378 lower limbs was 83.2 +/- 11.4 cm with a tibiofemoral ratio of 1 to 1.26 +/- 0.1. The 99th percentile level for length difference in 178 paired femora was 1.2 cm, in 171 paired tibiae 1.0 cm and in 60 paired lower limbs 1.4 cm. In 505 femora the mean internal torsion was 24.1 +/- 17.4 degrees, and in 504 tibiae the mean external torsion was 34.9 +/- 15.9 degrees. For 352 lower limbs the mean external torsion was 9.8 +/- 11.4 degrees. The mean torsion angle of right and left femora in individuals did not differ significantly, but mean tibial torsion showed a significant difference between right (36.46 degrees of external torsion) and left sides (33.07 degrees of external torsion). For the whole legs torsion on the left was 7.5 +/- 18.2 degrees and 11.8 +/- 18.8 degrees, respectively (p < 0.001). There was a trend to greater internal torsion in femora in association with an increased external torsion in tibiae, but we found no correlation. The 99th percentile value for the difference in 172 paired femora was 13 degrees; in 176 pairs of tibiae it was 14.3 degrees and for 60 paired lower limbs 15.6 degrees. These results will help to plan corrective osteotomies in the lower limbs, and we have re-evaluated the mathematical limits of differences in length and torsion. PMID- 9393925 TI - The effects of mechanical forces on bones and joints. Experimental study on the rat tail. AB - We have used an experimental model employing the bent tail of rats to investigate the effects of mechanical forces on bones and joints. Mechanical strain could be applied to the bones and joints of the tail without direct surgical exposure or the application of pins and wires. The intervertebral disc showed stretched annular lamellae on the convex side, while the annulus fibrosus on the concave side was pinched between the inner corners of the vertebral epiphysis. In young rats with an active growth plate, a transverse fissure appeared at the level of the hypertrophic cell layer or the primary metaphyseal trabecular zone. Metaphyseal and epiphyseal trabeculae on the compressed side were thicker and more dense than those of the distracted part of the vertebra. In growing animals, morphometric analysis of hemiepiphyseal and hemimetaphyseal areas, and the corresponding trabecular bone density, showed significant differences between the compressed and distracted sides. No differences were observed in adult rats. We found no significant differences in osteoclast number between compressed and distracted sides in either age group. Our results provide quantitative evidence of the working of 'Wolff's law'. The differences in trabecular density are examples of remodelling by osteoclasts and osteoblasts; our finding of no significant difference in osteoclast numbers between the hemiepiphyses in the experimental and control groups suggests that the response of living bone to altered strain is mediated by osteoblasts. PMID- 9393926 TI - Management of gunshot wounds of the limbs. PMID- 9393927 TI - Radio-opaque agents in bone cement increase resorption. PMID- 9393928 TI - Thromboprophylaxis and death after total hip replacement. PMID- 9393929 TI - Trauma management. PMID- 9393930 TI - Recurrent rotational deformity of the femur after static locking of intramedullary nails. PMID- 9393931 TI - Physical examination is sufficient for the diagnosis of sprained ankles. PMID- 9393932 TI - Management of fibular hemimelia. PMID- 9393933 TI - Articular penetration in Garden-I fractures of the hip. PMID- 9393934 TI - Rat natriuretic peptide receptor genes are regulated by glucocorticoids in vitro. AB - The effects of glucocorticoids on NPR-A and NPR-B mRNA transcription and natriuretic peptides ANP and CNP mediated cGMP production by intact vascular smooth muscle cells (VSMC) were studied in rat. Cultured VSMC were prepared from rat mesenteric arteries of 12-week-old Sprague-Dawley rats by enzymatic digestion. Dexamethasone-induced NPR-A mRNA increase was detectable early in the incubation periods and reached a plateau after 48 hours of glucocorticoid administration. This mRNA increase was mimicked by cortisol and inhibited by the glucocorticoid receptors antagonist RU 38,486. The levels of NPR-B mRNA remained unchanged during all the periods of stimulation. However, cGMP generated by both receptors in dexamethasone treated cells was higher than in control cells and this production was mimicked by cortisol and also blocked by RU 38,486. Desoxycorticosterone acetate (DOCA) had no effect on the levels of cGMP production. The results suggest that glucocorticoids have transcriptional and posttranscriptional effects on rat mesenteric arteries cells through glucocorticoid receptors. PMID- 9393935 TI - Prevention of hypoxemia-induced renal dysfunction by perindoprilat in the rabbit. AB - The role of angiotensin II, a potent postglomerular vasoconstrictor, in the hypoxemia-induced renal changes is still controversial. The ability of perindoprilat, an angiotensin converting-enzyme inhibitor, to prevent the acute renal effects of hypoxemia was assessed in 22 anesthetized-ventilated rabbits. In 8 untreated rabbits, hypoxemia induced a significant drop in mean blood pressure (MBP) (-12 +/- 2%), glomerular filtration rate (GFR) (-16 +/- 3%) and renal blood flow (RBF) (-12 +/- 3%) with a concomittant increase in renal vascular resistance (RVR) (+18 +/- 5%) and urine flow rate (+33 +/- 14%), and without any changes in filtration fraction (FF) (-4 +/- 2%). This suggests the occurrence of glomerular vasoconstriction during the hypoxemic stress. In 7 normoxemic rabbits, intravenous perindoprilat (20 microg/kg) induced an increase in urine flow rate (+17 +/- 4%) and RBF (+17 +/- 4%), and a decrease in MBP (-6 +/- 1%), RVR (-14 +/ 3%) and FF (-11 +/- 2%) without a significant change in GFR. The drop in FF and the increase in RBF suggests preferential postglomerular vasodilatation. In 7 rabbits, perindoprilat prevented the occurence of the hypoxemia-induced changes in RBF and RVR without improving MBP. FF decreased significantly (-18 +/- 2%), while the drop in GFR (-7 +/- 2%) was partially blunted and the increase in urine flow rate (+25 +/- 9%) was confirmed. These results could be explained by the inhibition of the angiotensin-mediated efferent vasoconstriction and by the inhibition of bradykinin degradation by perindoprilat. These data confirm the ability of converting-enzyme inhibitors to prevent the renal hypoperfusion induced by acute hypoxemia. PMID- 9393936 TI - Spinal mu, delta and kappa opioids alter chemical, mechanical and thermal sensitivities in amphibians. AB - Previously we demonstrated the use of chemical (topical acetic acid), thermal (radiant heat) and mechanical (von Frey filament) stimuli as quantifiable behavioral response assays in the northern grass frog, Rana pipiens. Furthermore, response thresholds in all of these sensory modalities are significantly elevated by systemic morphine injections, which can be antagonized by naltrexone. The present study employed these three sensory assays to assess changes in chemical, mechanical and thermal sensitivities following spinal administration of mu, delta and kappa opioids. Significant elevations were observed across all three sensory modalities in each subtype category and these effects were abolished by prior systemic administration of naltrexone. However, naltrexone antagonism of morphine produced hyperalgesia in both the mechanical and thermal modalities. The results support other recent work demonstrating that the spinal site for opioid analgesia is present in amphibians and that the thermal, mechanical and acetic acid assays are measures of true nociceptive activity in the amphibian. PMID- 9393937 TI - Antagonism to noradrenaline-induced lethality in rats is related to affinity for the alpha1A-adrenoceptor subtype. AB - The potency of several alpha1-adrenoceptor antagonists in preventing the noradrenaline-induced lethality in conscious rats, their binding affinity for the native alpha1A- and alpha1B-adrenoceptors, the recombinant animal alpha1a-, alpha1b- and alpha1d-adrenoceptor subtypes, as well as their functional affinity for the alpha1L-adrenoceptor subtype were evaluated. The potency of the tested compounds as antagonists of noradrenaline-induced lethality was correlated with the affinity for the alpha1A- (and alpha1a-) adrenoceptor subtype, but not with the affinity for the other subtypes. On the contrary, the hypotensive effects of the compounds, assessed in anesthetized rats, were not clearly related with the affinity for any of the alpha1-subtypes. These results suggest that the alpha1A subtype plays a determining role in preventing lethality induced by noradrenaline in the rats, and that this activity is unrelated to the hypotensive effect of the compounds, which cannot be clearly correlated with affinity for a particular alpha1-adrenoceptor subtype. PMID- 9393938 TI - The changes in pulmonary C-fiber activity and lung mechanics induced by vagal stimulation in rabbits. AB - The effect of unilateral stimulation of the vagus nerve on pulmonary C-fiber activity, total lung resistance (R(L)) and dynamic lung compliance (Cdyn) was studied in anesthetized, artificially ventilated rabbits. Vagal stimulation (4-30 Hz) increased both pulmonary C-fiber activity and R(L) but decreased Cdyn, and these changes were frequency-dependent. Atropine (2 mg/kg) blocked the responses of pulmonary C-fiber, R(L) and Cdyn to vagal stimulation at 4-30 Hz. Stimulation of pulmonary C-fiber activity by the higher volume-induced lung inflation was not significantly altered by atropine (2 mg/kg). A selective SP antagonist [D-Pro2, D Try(7,9)]-SP (0.5 mg/kg) inhibited bronchoconstrictor responses to vagal stimulation at 30 Hz but had no significant effect on those to the stimulation at 4-17 Hz. These results suggest that excitation of the rabbit pulmonary C-fiber activity by unilateral vagus nerve stimulation would occur as a result of the mechanical stimuli via a cholinergic neurotransmission and that during the stimulation at 30 Hz this excitation partly involves the local release of substance P (SP) to promote acetylcholine outflow. PMID- 9393940 TI - Role of enzymatic activity in the antiplatelet effects of a phospholipase A2 from Ophiophagus hannah snake venom. AB - A phospholipase A2 (OHV A-PLA2) from the venom of Ophiophagus hannah (King cobra) is an acidic protein exhibiting antiplatelet activity. In in vitro tests, OHV APLA2 showed a marked inhibitory effect on platelet aggregation induced by ADP, collagen and arachidonic acid in both human whole blood and platelet-rich plasma in a dose-dependent manner. The antiplatelet effects of OHV A-PLA2 did not increase when preincubation times of platelets and OHV A-PLA2 were prolonged indicating phospholipid hydrolysis did not significantly contribute to the antiplatelet effects. Alkylation of active site His residue using p-bromophenacyl bromide resulted in the complete loss of enzymatic activity, but the modified enzyme retained more than 30% of its antiplatelet effects. These results indicate that the antiplatelet effects of OHV A-PLA2 appear to be independent of its enzymatic activity, and there are separate sites responsible for the catalytic and antiplatelet activities. PMID- 9393939 TI - Nitric oxide synthase inhibition attenuates signs of ethanol withdrawal in rats. AB - The effects of N(G)-nitro arginine methyl ester (L-NAME) and 7-nitroindazole (7 NI), nitric oxide synthase inhibitors, and L-arginine, a nitric oxide precursor, on ethanol withdrawal signs were investigated in rats. Ethanol (7.2% v/v) was given to rats by a liquid diet for 16 days. L-NAME (30 and 60 mg/kg), 7-NI (40 and 80 mg/kg), L-arginine (100 mg/kg), a combination of L-arginine (100 mg/kg) and 7-NI (40 mg/kg), and saline or vehicle were injected to rats intraperitoneally 30 min before ethanol withdrawal. A second series of injections was given at 6 hour after the first one, and subjects were then tested for audiogenic seizures. 7-NI (40 mg/kg), vehicle and saline were also administered to naive rats. 7-NI (40 mg/kg) did not produce any significant change in locomotor activity in naive rats. Both L-NAME and 7-NI significantly inhibited locomotor hyperactivity from the 2nd to the 6th hour of the withdrawal period. They also reduced the total ethanol withdrawal score from the 30th min to the 6th hour, and they significantly decreased audiogenic seizures. Neither drug increased locomotor activity nor total ethanol withdrawal score, which were increased significantly by L-arginine (100 mg/kg); however, L-arginine (100 mg/kg) prevented the inhibitory effects of 7-NI (40 mg/kg) on increased locomotor activity, total ethanol withdrawal score, and audiogenic seizure. Our results suggest that nitric oxide synthase inhibition by L-NAME and 7-NI alleviates the signs of ethanol withdrawal. The data also support the hypothesis that nitric oxide may take part in the neuroadaptation that develops during chronic ethanol ingestion in rats. PMID- 9393941 TI - Amphetamine induces hydroxyl radical formation in the striatum of rats. AB - Amphetamine-induced hydroxyl radical formation in the striatum of rats was investigated in this study. With the utilization of the microdialysis and HPLC ECD, the striatal dopamine (DA) release and the formation of 2,3-dihydroxybenzoic acid (2,3-DHBA), derived from the reaction of hydroxyl radicals (.OH) and salicylate in perfusion, were monitored and detected during desipramine and/or amphetamine (AMPH) administration. Our data revealed that after desipramine treatment AMPH injections not only amplified striatal DA release and 2,3-DHBA formation, but also intensified the stereotyped behaviors induced by AMPH. Furthermore, we discovered that alpha-methyl-para-tyrosine (alpha-MT) pretreatment prevented the onset of the above responses. In desipramine-treated rats, the tissue homogenization study demonstrated that a single dose of AMPH produced long-term depletion of striatal DA; this was not seen in saline-treated rats. Moreover, striatal DA depletion could be lessened by pretreatment with mannitol, a .OH scavenger. These results indicate that AMPH-induced striatal .OH formation might be DA-related in desipramine-treated rats, and suggest that .OH formation might be correlated with AMPH-induced neurodegeneration. PMID- 9393942 TI - Iron binding capacity of didox (3,4-dihydroxybenzohydroxamic acid) and amidox (3,4-dihydroxybenzamidoxime) new inhibitors of the enzyme ribonucleotide reductase. AB - Ribonucleotide reductase is the rate limiting enzyme of deoxynucleoside triphosphate synthesis and is considered to be an excellent target of cancer chemotherapy. Didox and amidox are newly synthesized compounds, which inhibit this enzyme and have in vitro and in vivo antitumor activity. We have now investigated the capability of didox and amidox to interfere with the iron metabolism. We show by photometric and polarographic methods, that didox and amidox are capable of forming an iron complex. However, their cytotoxic action cannot be completely circumvented by addition of Fe-ammoniumcitrate, indicating that the iron complexing capacity may not be responsible for the mechanism of action of these compounds. When L1210 leukemia cells were incubated with the didox-iron or amidox-iron complex itself, changes of the 50% growth inhibitory capacity of the complex in comparison with didox or amidox alone could be shown. We conclude, that didox and amidox are capable of forming iron complexes, but in contrast to other agents, the anticancer activity cannot be contributed to this effect alone. Future studies will have to elucidate the molecular mechanism of action of these new and promising anticancer agents. PMID- 9393943 TI - With aging in humans the activity of the hypothalamus-pituitary-adrenal system increases and its diurnal amplitude flattens. AB - There is compelling evidence for feedback disturbances in the hypothalamus pituitary-adrenal system associated with human aging as assessed by challenge tests. However, reports about age-related changes in human basal activity are ambiguous and to date little is known about changes in the pulsatile features of the HPA system. To investigate these changes we studied twenty-two healthy male and eleven healthy female subjects ranging from 23 to 85 and 24 to 81 years respectively. 24-hour blood sampling with 30 minute sampling intervals was performed. From 18.00 to 24.00 hours blood was sampled every 10 minutes for analysis of pulsatile features of HPA activity. Statistical analysis revealed that age in particular had major effects upon basal HPA-system activity: there was a significant age-associated increase in minimal (p < 0.0001) and mean (p < 0.02) cortisol plasma concentrations, but no alteration in pulsatile features. We found no age-cortisol correlation during daytime, but were able to demonstrate a strong impact of age upon cortisol plasma levels from 20.00 to 1.30 hours. The diurnal amplitude of cortisol (p < 0.005) and ACTH (p < 0.006), relative to the 24-hour mean of the hormones, showed an age-associated decline. Additionally, the evening cortisol quiescent period (p < 0.01) was shortened in the elderly, suggesting increasingly impaired circadian function in aging. Our results suggest an increased basal activity and a flattened diurnal amplitude of the HPA system in the elderly. PMID- 9393944 TI - The effect of metabolic acidosis and alkalosis on the H+-ATPase of rat cerebral microvessels. AB - To determine the role of the proton translocating adenosine triphosphatase (H+ ATPase) of the blood-brain barrier, the density of the 31 Kd subunit of the vacuolar type H+-ATPase was quantitated in isolated rat cerebral microvessels with immunoblotting techniques. To establish the tissue specificity of the findings, synaptosomal membranes were also studied. Metabolic acidosis was induced with 1.5% ammonium chloride in drinking water for five days. Metabolic alkalosis was induced with 2.35% NaHCO3 in drinking water and daily injections of 10 mg/Kg furosemide intraperitoneally for 5 days. The quantity of the 31 Kd subunit (in arbitrary units) in cerebral microvessels was significantly increased in acidosis (3.98 +/- 0.45) (p<0.05) and was significantly decreased in metabolic alkalosis (0.49 +/- 0.16) (p<0.00) compared to controls (1.77 +/- 0.73). In synaptosomal membranes, metabolic alkalosis was associated with significant decrease in the quantity of the 31 Kd subunit-H+-ATPase (0.62 +/- 0.12 vs 0.92 +/ 0.01) p<0.05. The increase in the 31 Kd subunit in synaptosomal membranes with acidosis did not reach statistical significance. It is concluded that the quantity of vacuolar H+-ATPase in the blood-brain barrier is modulated by blood H+ or HCO3- content. These changes may be relevant to the physiology of the acid base balance in the central nervous system. PMID- 9393945 TI - Cyclosporine A-induced contraction of isolated rat aortic smooth muscle cells. AB - The mechanisms by which the immunosuppressive drug cyclosporine A (CsA) induces hypertension and nephrotoxicity are still not fully understood. Although smooth muscle cell (SMC) contraction is probably the mechanism of vasoconstriction, the direct contractive effect of CsA on SMCs has not yet been demonstrated. Thus, it was the purpose of this study to evaluate the direct effects of CsA in cultured SMCs through interactive image analysis. In aortic SMCs, CsA at the concentrations of 0.01, 0.1 and 1 microM, caused a concentration-dependent decrease of the planar cross-sectional area (PCSA) after 30 min and 60 min of treatment. The PCSA decreases were statistically significantly different from control at all concentrations. No cytotoxicity was observed under these conditions. Ten minutes preincubation of SMCs with a monoclonal antibody against endothelin-1 (ET-1) significantly prevented the CsA effects at 1 microM. When the same antibody was heat inactivated or an unspecific antibody (anti-desmin immunoglobulin G) was applied, the CsA-induced contractions were not affected. These data suggest that CsA can cause a direct contractive effect on vascular SMCs. This effect is partly mediated by ET-1. PMID- 9393946 TI - Characterization of methadone receptor subtypes present in human brain and lung tissues. AB - In addition to their use in pain control, opioids can function as regulators of tumor cell growth. We have found that the therapeutic opioid, methadone, significantly inhibits the in vitro and in vivo growth of human lung cancer cells, and this effect appears to be mediated by specific, high affinity, non conventional opioid binding sites. The present study indicates the existence of multiple subtypes of binding sites mediating the peripheral and central nervous system actions of this drug. Pharmacological and biochemical characterizations of the methadone binding sites expressed in human brain and normal lung tissues indicate that these sites are distinct from each other and from other opioid receptor types present on human and rat brain membranes, as well as those expressed in human lung cancer cells. The identification of distinct methadone receptor types in the different tissues could lead to the development of more selective and less toxic drugs targeted toward the tumor cells. PMID- 9393947 TI - The role of estrogen receptor mutations in tamoxifen-stimulated breast cancer. AB - During the past 20 years, the hormonal therapy of choice for the treatment of breast cancer has been the antiestrogen, tamoxifen. The use of tamoxifen has been proved to produce a favorable response and survival advantage in patients whose tumors are classified as estrogen receptor-positive (ER+)/progesterone receptor positive (PR+). Additionally, tamoxifen is the only drug known to reduce the incidence of contralateral disease. This drug produces relatively few harmful side effects, while exhibiting several beneficial effects such as maintaining bone density and reducing the incidence of myocardial infarction in the postmenopausal woman. However, tumors eventually acquire a tamoxifen-resistant or tamoxifen-stimulated phenotype, resulting in disease recurrence. Several mechanisms have been proposed to account for tamoxifen-resistant breast cancer, in the hope of developing a more effective first-line or perhaps second-line treatment strategy. One popular theory is the occurrence of a mutation in the estrogen receptor, the drug target. A plethora of studies have reported the detection of estrogen receptor mRNA splice variants, and it has been suggested that the accumulation of these variant mRNAs are responsible for the development of tamoxifen-resistant breast cancer. In this review, several questions will be posed to address the suitability of both laboratory and clinical evidence to support this hypothesis. Although there is adequate data generated in the laboratory, there is, as yet, no compelling evidence to suggest that mutation of the estrogen receptor is the molecular mechanism producing tamoxifen-stimulated growth in human breast and endometrial cancer. PMID- 9393948 TI - Trans-retinoic acid and glucocorticoids synergistically induce transcription from the mouse mammary tumor virus promoter in human embryonic kidney cells. AB - Human embryonic kidney (K293) cells transfected with a mouse mammary tumor virus (MMTV) promoter-luciferase reporter construct (pHH-Luc) were utilized to investigate the potential effects of trans-retinoic acid (tRA), either by itself or in combination with glucocorticoid (GC) hormones, on a well-characterized, GC sensitive transcriptional response. tRA or the synthetic GC hormone dexamethasone induced transcription from the MMTV promoter in a dose-dependent manner, with 1 micromol tRA and 1 micromol dexamethasone alone causing a four- to six-fold and a 40-fold induction of basal transcription, respectively. Simultaneous treatment with 1 micromol dexamethasone and 1 micromol tRA resulted in a synergistic transcriptional response that was 120-fold higher than basal level and 2.5 times the predicted response, based on a simple additive effect of both agonists. tRA does not appear to mediate this synergistic transcriptional response by enhancing GC receptor (GR) binding capacity, affinity, or nuclear translocation. tRA was unable to potentiate GC-induced transcriptional activity from a minimal GC response element (GRE), and GC were unable to potentiate tRA-induced transcriptional activity from a minimal retinoic acid response element (RARE). These data rule out direct protein-protein interactions between GC and retinoid receptors as a mechanism for the observed synergism. tRA also synergized with aldosterone-induced, mineralocorticoid receptor (MR)-mediated, transcriptional activation of the MMTV promoter, resulting in a response that was 1.7 times the predicted additive response. The MMTV GRE located between -187 and -165 was required for GC-induced and synergistic activation of the MMTV promoter, whereas sequences located within -151 to +5 were sufficient for tRA-induced transcription from the MMTV promoter. Mutation of a consensus RARE half-site (CCAAGT) identified at position -65 to -60 within the MMTV-LTR did not affect either tRA induced transcriptional activation or synergism with GC. We propose that the tRA induced transcriptional response from the MMTV promoter, as well as synergism with GC, may be mediated by the activation or induction of a factor(s) that either directly binds to the MMTV promoter or indirectly stabilizes binding of another transcription factor to these sequences. PMID- 9393949 TI - Identification of two estrogen receptor transcripts with novel 5' exons isolated from a MCF7 cDNA library. AB - Two novel transcripts of human estrogen receptor (ER) have been identified that differ in the 5' untranslated sequence. It has previously been determined that an alternate ER transcript is generated from transcription initiated upstream of the main ER cap site (P1), and utilizes a splice acceptor site at +163. Here we report the isolation of 21 ER clones from a MCF7 cDNA library. Eleven of these clones correspond to transcripts that initiate at the P1 cap site, whereas the remaining 10 clones are derived from two previously unidentified ER transcripts (designated E and H) that both utilize the +163 splice acceptor site. A panel of breast and endometrial carcinoma cell lines were screened by reverse transcriptase-polymerase chain reaction (RT-PCR) for expression of the E and H transcripts. It was found that all ER-positive cell lines expressed both of the novel transcripts. In addition, 10 primary human breast cancers were analyzed, of which six expressed the E transcript and five abundantly expressed the H transcript. These data indicate that expression of ER in human breast cancers can be dependent upon an alternate promoter at least 20 kb upstream of the primary cap site for ER. PMID- 9393950 TI - Auto-regulation of the estrogen receptor promoter. AB - The presence or absence of estrogen receptor (ER) plays a key role in the diagnosis and treatment of breast tumors. It is known that patients with breast tumors classified as ER-positive have a better prognosis. Observations such as this have led us to explore the question of what makes some breast tumors overexpress ER whereas others express either very low levels or none at all. To begin a study of ER regulation, we first chose to examine a 200 bp region of the ER promoter located immediately upstream from the transcribed sequence of the human ER gene. We found that this region of the ER promoter contained basal activity when transiently transfected into ER-negative HeLa cells. ER promoter activity was further increased by co-transfection of a wild-type ER expression vector, and this increased activity was hormone-dependent. Several ER deletion mutant constructs were also able to increase the activity of the ER promoter fragment, but none could support equivalent activity as was seen with the full length ER. Therefore, we conclude that the ER can contribute to its own expression, and we hypothesize that this auto-regulation may contribute to its overexpression in some breast tumors. PMID- 9393951 TI - A reporter gene assay for fungal sterol biosynthesis inhibitors. AB - Acetoacetyl-CoA thiolase (ACoAT) catalyses the condensation of two acetyl-CoA molecules, the first step in the sterol biosynthetic pathway. We constructed a yeast strain containing a fusion of the promoter of the Saccharomyces cerevisiae ACoAT gene to a reporter gene (Escherichia coli beta-galactosidase). Reporter gene activity in this strain can be induced by a variety of inhibitors of sterol biosynthesis. These results suggest that the ACoAT gene is feedback regulated at the transcriptional level by products of the sterol biosynthetic pathway. The reporter gene approach described here may be used to screen chemical collections for compounds which inhibit fungal sterol biosynthesis. PMID- 9393952 TI - Neutral mutations to three acidic AF2 residues in the mouse estrogen receptor confer agonist activity to A-ring isomers of estradiol. AB - The ability of the estrogen receptor (ER) to function as a ligand-mediated transcription factor involves the activation function-2 (AF2) in the hormone binding domain (HBD). Although several types of ligand bind to the ER, AF2 functions selectively, as it is activated by estradiol (E2) but not by antiestrogens. The mechanism used by AF2 to interpret the chemical and structural information encoded in the bound ligand, and to transfer this information to other transcriptional regulatory proteins on the promoters of estrogen-regulated genes, is unknown at present. To address this issue, we have examined the activities of two mouse ERs with mutations in the AF2 region. One incorporated changes in three acidic residues (pJ3MOR, D542N/E546Q/D549N) on the polar face of the putative AF2 alpha-helix, whereas the other contained alterations in two hydrophobic amino acids (pJ3MOR, L543D/L544A) on the non-polar face. Transcriptional activity was measured with chloramphenicol acetyltransferase (CAT) reporter genes including a minimal (JA12) or a complex (pS2tkCAT) promoter. In transient cotransfection assays using COS-1 cells, it was found that A-ring isomers of E2, which are inactive or behave as antagonists with the wild-type ER, acted as agonists when the neutral AF2 mutant ER and the pS2tkCAT promoter were tested. On the other hand, when the mutant ER with changes in key hydrophobic residues was used with this same promoter, the estrogen antagonist, ICI 164,384, acted as an agonist. The findings in this report establish a role for acidic AF2 amino acids, and confirm the contributions made by hydrophobic residues, in the interpretation of ligand identity and in the transmission of this information to the transcriptional regulatory apparatus. Critical residues on both sides of the AF2 alpha-helix are, therefore, likely to be involved in distinguishing between ligands with either extensive or subtle alterations in structure, and in mediating this information to the regulatory proteins on estrogen-responsive genes. PMID- 9393953 TI - The effect of chorion-uterine interaction upon free progesterone metabolism during advanced gestation in the guinea pig. AB - The in vitro fate of [3H]progesterone was studied after incubation with guinea pig tissues at 58/59 days (before pubic symphysis relaxation) and in the final week (post relaxation) of gestation. Buffered steroid was added to the fetal surface of chorion attached to the uterus or to the uterus alone. Neither the amount of recovered progesterone nor its metabolites (6.2% average conversion) differed between the two stages when only uterus was incubated. With chorion present, conversion averaged 28.3% at 58/59 days and 63.4% at the late stage. A 4.6-fold decrease in progesterone concentration, and 3.0-, 2.4- and 3.1-fold increases in the concentrations of 3alpha-hydroxy-5alpha-pregnan-20-one, 3beta hydroxy-5alpha-pregnan-20-one and 5alpha-pregnane-3,20-dione, respectively, were found in the uterus in the late stage vs 58/59 days. Also, 2.8- and 6.4-fold decreases in progesterone concentration occurred in the myometrium and endometrium, respectively, from 58/59 days to the late stage. In endometrium, the concentrations of the 3alpha- and 3beta-isomers, and 5alpha-pregnane-3,20-dione, increased 2.6-, 2.6- and 5.0-fold, respectively. The above changes were all significant at P < 0.05 or better. Changes in the 3alpha-, 3beta- isomers and dione in the myometrium were not significant. The chorion-uterine interaction and gestation time thus affect the degree of progesterone conversion, and the amounts of metabolites reaching the uterus in the chorion-uterine in vitro system. PMID- 9393954 TI - Metyrapone is a competitive inhibitor of 11beta-hydroxysteroid dehydrogenase type 1 reductase. AB - The present study was designed to examine the effects of metyrapone in vitro on the activities of 11beta-hydroxysteroid dehydrogenase (11beta-HSD) types 1 and 2, the two intracellular enzymes responsible for the metabolism of glucocorticoids. Enzymatic activities of 11beta-HSD1 and 2 were determined by a radiometric conversion assay using cortisol and cortisone as physiological substrates. The enzyme activity assays were carried out in the absence and presence of metyrapone using sheep liver and kidney microsomes as the source of 11beta-HSD1 and 2, respectively. It was found that metyrapone inhibited the reductase activity of 11beta-HSD1 in a dose-dependent manner with an apparent Ki of 30 microM. Moreover, this inhibition was competitive because the Km for cortisone was increased in the presence of metyrapone. In contrast, metyrapone showed biphasic effects on the dehydrogenase activity of 11beta-HSD1, in that it increased the activity at concentrations lower than 100 microM but decreased it at higher concentrations. However, under similar conditions, metyrapone had little effect on the unidirectional dehydrogenase activity of 11beta-HSD2. In conclusion, the present results provide the first direct evidence that metyrapone is a competitive inhibitor of 11beta-HSD1 reductase, and that it also exerts biphasic effects on 11beta-HSD1 dehydrogenase activity. These findings indicate that metyrapone influences peripheral glucocorticoid metabolism through its regulation of 11beta-HSD1 activity, in addition to its classic inhibitory effects on adrenal steroid biosynthesis. It is therefore imperative that this novel extra-adrenal effect of metyrapone be considered when this drug is used in the diagnosis and treatment of adrenocorticoid-related diseases. PMID- 9393955 TI - Second messengers and the control of progesterone production from first trimester trophoblast. AB - Basal progesterone production from first trimester placental cells in culture was high during the first 24 h of culture and fell to less than 30% of the initial level after 96 h in vitro. 22(R)-Hydroxycholesterol had a similar effect on progesterone production at all incubation times, indicating that the decline in basal steroidogenesis was not due to a loss of mitochondrial or post mitochondrial enzymes. Continuous stimulation with dibutyryl (db) cyclic AMP maintained progesterone synthesis at a relatively constant high level despite the fall in basal progesterone production, and the optimum concentration of db cyclic AMP was 1.0 mM. The calcium ionophore A23187 had no effect on progesterone incubation during short-term cultures (<4 h), and inhibited steroidogenesis after 24 h. Repeated addition of A23187 during 96 h of culture also inhibited progesterone production. These findings indicate that progesterone production in human trophoblast is supported by a local factor which maintains a high level of steroid production through a cyclic AMP-dependent mechanism. The inhibitory effects of calcium ionophore in trophoblast differ from the stimulatory effects of this compound in other steroidogenic cells, but the reasons for the difference are not known at present. PMID- 9393956 TI - Inhibition of steroidogenesis in rat adrenal cells by 18 ethynyldeoxycorticosterone: evidence for an alternative pathway of aldosterone biosynthesis. AB - The effect of the mechanism-based inhibitor 18-ethynyldeoxycorticosterone (18-E DOC) on the late steps of the aldosterone biosynthetic pathway was examined in freshly isolated cells of the zona glomerulosa (ZG) and fasciculata (ZF) from rat adrenal glands. ZG synthesis of aldosterone was inhibited by 18-E-DOC in a time- and concentration-dependent manner with a Ki of approximately 0.05 microM. The maximal degree of inhibition of ZG production of aldosterone and 18 hydroxycorticosterone (18-OH-B) was approximately 80%. ZF cells, perhaps surprisingly, were found to secrete 18-OH-B at levels approximately one-third to one-fourth those of ZG cells and the Ki of 18-E-DOC inhibition of 18-OH-B secretion was approximately 10 microM for ZF cells, 200-fold higher than for ZG cells. The inhibitor had no effect on the secretion of corticosterone by either ZG or ZF, and the secretion of 18-hydroxydeoxycorticosterone (18-OH-DOC) by both the ZG and ZF was inhibited only to a minor degree. 18-E-DOC inhibited the biosynthesis of aldosterone by ZG cells incubated with 10 microM added DOC or 18 OH-DOC by approximately 75%, similar to the degree of inhibition of aldosterone biosynthesis from endogenous substrate, whereas ZF biosynthesis of 18-OH-B from either substrate was inhibited by less than 40%. ZF cells do not express aldosterone synthase, the only enzyme known to convert 18-OH-DOC into 18-OH-B. Incubation of MA-10 cells stably transfected with the cDNA of the rat aldosterone synthase with 18-E-DOC resulted in a complete inhibition of the conversion of DOC to aldosterone with a Ki of approximately 0.02 microM. In addition, transfected cells expressing 11beta-hydroxylase convert DOC to 18-OH-B in very small quantities only and cannot convert 18-OH-DOC to 18-OH-B. These data suggest that neither 11beta-hydroxylase nor aldosterone synthase are responsible for the biosynthesis of 18-OH-B by ZF cells from DOC or 18-OH-DOC, that 20% of aldosterone synthesis appears not to be attributable to the actions of aldosterone synthase and that an unknown CYP11B enzyme is also involved in the biosynthesis of 18-OH-B. PMID- 9393957 TI - Steroid productions by co-cultures of granulosa cells with inner and outer theca cells in preovulatory follicles of gonadotropin stimulated calves. AB - Granulosa, interna and externa theca cells were isolated from large follicles of equine-chorionic-gonadotropin (eCG)-primed calves and co-cultured during 3 days in the absence or in the presence of dehydroepiandrosterone (DHEA). Co-cultures were performed by adding defined numbers of theca and/or granulosa cells which represented 0, 10, 20, 50 or 100% of total cells per well. Secretion of oestradiol-17beta (E2), androstenedione (A4) and progesterone (P4) depended on the type of theca cells (P < 0.001), on the percentage of seeded granulosa cells (P < 0.001) and on the day of culture (P < 0.001). DHEA increased (P < 0.001) E2 and A4, but not P4 (P > 0.05) productions. Interactions existed between these factors (P < 0.01). On day 1, A4 production was nil in granulosa cells alone. E2 production was negligible in theca cells alone but it increased when granulosa cells were added. E2 and A4 varied in an opposite manner according to the percentage of granulosa cells and with the type of theca cells. On day 3, without DHEA, E2 and A4 were low. On day 3 with DHEA, E2 production was maintained in granulosa cells alone but not with any combination of theca cells. In these conditions, A4 production was maintained in the presence of theca cells but not in granulosa cells alone. Granulosa cells alone secreted more P4 than theca cells. P4 increased as a function of the percentage of granulosa in co-cultures with externa but not interna theca cells with which it remained low. In conclusion, theca cells in culture have two effects in relation to the granulosa cells, which differ according to the steroid concerned and to the cell combination. Both types of theca cells have an inhibitory effect on E2 secretion whereas only interna theca cells are able to alter P4 production. PMID- 9393958 TI - Induction of cytochrome P450 1B1 and catechol estrogen metabolism in ACHN human renal adenocarcinoma cells. AB - The catechol estrogen metabolites of 17beta-estradiol (E2), 2-hydroxyestradiol (OHE2) and 4-OHE2, differ in hormonal properties and carcinogenic potential. In Syrian hamster kidney, 4-OHE2 induces clear-cell carcinoma whereas 2-OHE2 does not, and an E2 4-hydroxylase appears to be involved in E2-induced carcinogenesis in these animals. Specific E2 4-hydroxylase activity has been observed in extrahepatic tissues from several species. In humans, cytochrome P450 1B1 (CYP1B1) appears to be an extrahepatic E2 4-hydroxylase under the regulatory control of the aromatic hydrocarbon receptor (AhR). As an initial approach to investigating CYP1B1 expression and E2 4-hydroxylase activity in human kidney, we used the ACHN cell line, derived from a human renal adenocarcinoma. In untreated ACHN cells, a very low level of CYP1B1 mRNA expression was observed and CYP1B1 protein could not be detected; however, in ACHN cells exposed to the high affinity AhR ligand, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), CYP1B1 mRNA levels were elevated 28-fold, and the CYP1B1 protein was detected by immunoblot analysis. Exposure of ACHN cells to TCDD resulted in minimal induction of the CYP1A1 mRNA, and the CYP1A1 protein was not detectable prior to or after exposure to TCDD. E2 hydroxylase activity could not be detected with microsomes from untreated ACHN cells, although activities at C-4 and, to a lesser extent, at C-2 of E2 were observed with microsomes from TCDD-treated ACHN cells. In experiments with intact ACHN cells, elevated rates of formation of 4-methoxyestradiol (MeOE2) and 2-MeOE2 were observed in response to treatment with TCDD. The EC50 for induction of the CYP1B1 mRNA was 1.5 nM TCDD; EC50s for the stimulation of 2- and 4-MeOE2 formation were 0.68 and 1.1 nM TCDD. These results indicate that the ACHN cell line may be a useful in vitro model system to study the regulation of CYP1B1 expression and the cytotoxic effects associated with E2 4-hydroxylation. PMID- 9393960 TI - Identification and characterization of polymorphisms in the promoter region of the human Apo-1/Fas (CD95) gene. AB - Apo-1/Fas (CD95) is a transmembrane protein expressed on the cell surface that is involved in apoptosis and plays an important role in the function and regulation of the immune system. Aberrant expression of the Apo-1/Fas gene product has been reported in a number of immune-related disorders, such as autoimmune lymphoproliferative syndrome and systemic lupus erythematosus in humans. Mutations in the coding sequence of the Apo-1/Fas gene have been reported in the former condition, whereas no abnormalities of the gene have been found to account for the increased gene expression noted in SLE. We screened the whole 5' flanking region of the Apo-1/Fas gene encompassing over 2000 bp for mutation(s)/polymorphism(s) using multiplex PCR, single-strand conformation polymorphism (SSCP) analysis and sequencing techniques, and identified two polymorphisms in this region. The first polymorphism is a CG-->CA substitution at -1377 nucleotide position within the silencer region, which neither creates or deletes any restriction enzyme sites but alters the transcription factor SP-1 binding site. This polymorphism is noted in 20% of normal Caucasians. The second polymorphism is an GA-->GG substitution at -670 nucleotide position in the enhancer region that creates a MvaI restriction fragment length polymorphism (RFLP) and abolishes the binding site of nuclear transcription element GAS. The MvaI RFLP is polymorphic with heterozygosity of 52% and the frequency of G and A alleles are 0.49 and 0.51, respectively. The identification and characterisation of these two new polymorphisms, particularly the MvaI RFLP marker, provides new genetic markers and may prove useful for further studies on the regulation of apoptosis mediated by the Apo-1/Fas gene on human chromosome 10q23. PMID- 9393959 TI - Synthetic lipopeptides incorporated in liposomes: in vitro stimulation of the proliferation of murine splenocytes and in vivo induction of an immune response against a peptide antigen. AB - Amphiphilic lipopeptides, such as Pam3CysAlaGly and Pam3CysSerSer, were synthesized and incorporated into liposomes, and their ability to induce the proliferation of BALB/c mouse splenocyte was tested in vitro. When compared to monophosphoryl lipid A (MPL) the following potency order was found: liposomal lipopeptides > liposomal MPL > free (emulsified) lipopeptides. These results strongly depend on the size of the vesicles used: a mitogenic effect was observed only with lipopeptides incorporated within vesicles of diameter < or = 100 nm while lipopeptides in larger vesicles (diameter approximately 300 nm) gave no response. This may be related to the necessity for the liposome-associated lipopeptides to be endocytosed to reach putative intracellular targets. As immunoadjuvanticity seems to be linked to B-lymphocyte activation, the lipopeptides represent attractive alternatives to MPL for the realization of completely synthetic liposome-based peptide vaccine formulations. This was borne out by showing that Pam3CysAlaGly and Pam3CysSerSer, when incorporated in small unilamellar vesicles carrying a covalently conjugated synthetic peptide of sequence IRGERA, corresponding to an epitope of the C-terminal region of histone H3, were able to induce a potent and long-lasting immune response. PMID- 9393961 TI - Isolation and N-terminal sequence determination of a novel gamma/delta T cell surface antigen. AB - An antigen complex unique for porcine gamma/delta T cells has previously been identified using the monoclonal antibodies MAC319 and MAC320. Here we use digestion with the proteolytic enzyme bromelain to selectively release the MAC319 antigen as a soluble fragment, for further characterisation. A cytofluorometric inhibition assay was developed to follow the purification of this fragment, as the conformation sensitivity of the MAC319 epitope prevented the use of immunoblotting techniques. The antigen has been purified using a combination of anion-exchange and hydrophobic-interaction columns, followed by separation on a size-exclusion column. Fractions from the size-exclusion column containing the antigen consisted of one major band at Mr 95,000. This species was shown to be specifically absorbed onto MAC319-coupled Sepharose, thereby identifying the MAC319 antigen. N-terminal amino acid sequencing of this band has revealed a previously unidentified sequence. This fragment was also shown to be glycosylated, most likely with a single sugar moiety. Enzymatic removal of the sugars showed that they did not appear to be necessary for binding of the polypeptide to the MAC319 antibody. PMID- 9393962 TI - Glycosylation is influential in murine IgG3 self-association. AB - In mice and humans, antibodies of the IgG3 isotype are unique in that they spontaneously self-associate. A consequence is the formation of cryoglobulins from some, but not all IgG3 molecules. Little is known about the structural basis of murine IgG3 self-association. A region of the CH3 domain that is unique to IgG3 antibodies is the presence of an extra glycosylation site at residues 471 473. It is known that glycosylation greatly influences solubility. It has also been shown by X-ray crystallography that glycosylated residues (specifically sialic acid) are influential in the contacts of the CH1 to CH2 as well as the CH2 to CH2 domains in a human IgG1 antibody. These findings provided evidence that a direct interaction exists between the glycosylated residues and other residues within the constant and/or variable domains. It was, therefore, important to determine whether the glycosylated residue in the CH3 domain of the IgG3 constant region is influential in self-association. We have found that removing the glycosylation site by site-directed mutagenesis of an IgG3 RF significantly reduced the self-associating ability of this antibody. PMID- 9393963 TI - Tandem repeats of T helper epitopes enhance immunogenicity of fusion proteins by promoting processing and presentation. AB - Empirical findings have shown that recombinant chimeric proteins may be made more immunogenic if T helper epitopes are incorporated as tandem repeats. In the present study we investigated the mechanisms responsible for the enhanced immunogenicity of fusion proteins composed of the heat-stable enterotoxin of enterotoxigenic E. coli (STa) linked to multiple copies of the ovalbumin323-339 T helper epitope (ova) and a connecting dimer of an Ig-binding region of Staphylococcus aureus protein A (ZZ), which were previously shown to stimulate strong anti-STa titres in mice. We used B cell and macrophage cell lines as APC and IL-2 production by ova-specific T cells as our read-out system. Fusion proteins containing four repeated T helper epitopes were found to be the most immunogenic and resulted in 50-fold higher IL-2 production than constructs with a single T helper epitope. Under limiting APC conditions the construct with four epitopes was the best inducer of IL-2, indicating that this construct was most effectively processed by the APC. Analysis of IL-2R alpha expression by flow cytometry confirmed that four copies gave the highest frequency of activated T cells in culture, indicating a direct correlation between ability to activate T cells and IL-2 production in culture. Also in vivo, the fusion protein with four epitopes exhibited the strongest T cell priming effect. Moreover, both in vitro and in vivo, the ZZ construct was found to serve as an efficient means for targeting of the fusion proteins to B cells, thereby allowing access to the Ig receptor uptake pathway for Ag. The present study provides direct evidence that fusion proteins can be constructed to optimize processing in the individual APC and enhance activation of clonal T cells. PMID- 9393964 TI - Cloning of anti-Gal Fabs from combinatorial phage display libraries: structural analysis and comparison of Fab expression in pComb3H and pComb8 phage. AB - Anti-Gal is the most abundant natural antibody in humans. It interacts specifically with the carbohydrate epitope Gal alpha1-3Galbeta1-4GlcNAc-R (termed the alpha-galactosyl epitope). In an attempt to characterize the Ig genes encoding anti-Gal, two combinatorial phage display libraries in phagemid pComb3H were screened for anti-Gal Fabs. For this purpose, phages were incubated with biotinylated BSA coupled with alpha-galactosyl epitopes (designated alpha-Gal BSA). Subsequently, phages complexed with alpha-Gal-BSA were isolated by streptavidin-coupled magnetic beads. Because of the low affinity of this antibody, a characteristic shared with other anti-carbohydrate antibodies, only two clones displaying anti-Gal activity were isolated. Clone G9 contained the VH gene V3-43 and VL gene DPK15, whereas clone P19 contained the VH gene V3-15 and VL gene DPL16. Both clones contained between five and 14 mutations in their H and L chain V genes. The affinity of clone G9 was found to be higher than that of clone P19, as only the former could bind to solid-phase alpha-galactosyl epitopes in an enzyme-linked immunosorbent assay. This interaction could be increased by expressing Fabs in phagemid pComb8 grown in the presence of IPTG. Under such conditions, the IPTG-activated Lac-Z promoter induces an increased expression of Fabs that are linked to phage envelope protein VIII, resulting in multiple Fab display on the phage. The data suggest that screening combinatorial phage display libraries for anti-carbohydrate antibodies may be more effective with pComb8 phage grown in the presence of IPTG. PMID- 9393965 TI - Molecular cloning, expression and characterization of Pru a 1, the major cherry allergen. AB - A high percentage of birch pollen allergic patients experiences food hypersensitivity reactions after ingestion of several fruits and vegetables. Previous work demonstrated common epitopes on an allergen of Mr 18,000 from sweet cherry (Prunus avium) and Bet v 1, the major allergen from birch pollen. N terminal amino acid sequencing showed a sequence identity of 67% with Bet v 1. Here we report the cloning and cDNA sequencing of this cherry allergen. The entire deduced amino acid sequence described a protein of Mr 17,700 with 59.1% identity to Bet v 1. High degrees of identity in the range of 40 to 60% were also found with related allergens from other kinds of tree pollen and plant foods as well as with stress-induced proteins from food plants such as parsley, potato and soya. The coding DNA of the cherry protein was cloned into vector pET-16b and expressed in E. coli strain BL21(DE3) as a His-tag fusion protein. As shown by SDS-PAGE, the apparent molecular masses of the nonfusion protein and the natural allergen were identical. The fusion protein showed high IgE binding potency when sera from patients allergic to cherry were tested by immunoblotting and enzyme allergosorbent tests. Moreover, it cross-reacted strongly with IgE specific for the natural counterpart and for Bet v 1. The high biological activity of the recombinant fusion protein was further confirmed by the induction of a strong histamine release in basophils from cherry-allergic patients. Since sera from 17/19 of such patients contained IgE against this allergen it was classified as a major allergen and named Pru a 1. Recombinant Pru a 1 mimics most of the allergenic potency of cherry extract and hence could be a useful tool for studying the molecular and immunological properties of pollen related food allergens. PMID- 9393966 TI - Structure and organization of the immunoglobulin M heavy chain genes in Atlantic salmon, Salmo salar. AB - To determine the structure and organization of the germline immunoglobulin M heavy chain (IgH) genes in Atlantic salmon, Salmo salar, relevant clones from a genomic library (of one individual fish) have been characterized. Two closely related IgH constant region genes, CHA and CHB, have been sequenced completely. In addition, an allotypic variant of CHA was identified and partially sequenced. Five joining (JH) elements were found in a distance of 0.5-1.6 kb upstream of the first constant exon (CH1), in both CHA and CHB, substantiating the hypothesis that the entire gene complex is duplicated; possibly a remnant of a tetraploid event in the salmonid ancestor. An octamer motif (ATGTATTT, and its reverse complementary sequence) was found to be dispersed in the JH-CH1 region, but not elsewhere, signifying a role in these loci. Four closely related variable (VH) genes which were subcloned from three distinct lambda clones showed the classical structure of a two exon unit split by a 100 bp intron. The split-intron and a few hundred base pairs of the flanking sequences of the genes were highly similar. Three of the four genes were interrupted by stop codons and/or frame shifts, indicating a high proportion of VH-pseudogenes in this species. Based on the present results, and comparison with sequences of rainbow trout, Oncorhynchus mykiss, it is likely that the IgH loci have remained tetrasomicly inherited throughout the radiation of the genus Salmo and Oncorhynchus, and that the duplicated loci have gone into a disomic inheritance pattern in the comparatively recent past. PMID- 9393967 TI - A single predominantly expressed polymorphic immunoglobulin VH gene family, related to mammalian group, I, clan, II, is identified in cattle. AB - In order to understand the generation of antibody diversity in cattle, seven cDNAs, from heterohybridomas secreting bovine IgM and IgG1 antibodies, were cloned and structurally analyzed for rearranged bovine VDJ genes. All of the seven bovine VH genes, together with four available bovine VH gene sequences, shared a high nucleotide sequence homology (84.2-93.5%). Based upon the criteria of nucleic acid homology > or =80%, all of the bovine VH gene sequences isolated from the expressed antibody repertoire constitute a single VH gene family, which we have designated as bovine VH1 (Bov VH1). An analysis of 44 bovine IgM secreting mouse x cattle heterohybridomas, originating from polyclonally activated PBLs from bovine leukemia virus-infected cattle, revealed that all of these expressed Bov VH1 (100%) based upon DNA sequencing and Northern dot blot. The bovine VH genes showed highest DNA sequence similarity, ranging between 81.5 and 87.6%, with a single sheep VH gene family (related to human VH4) and are, thus, closest to the VH genes from another ruminant species. The Bov VH1 gene family is most homologous to the murine VH Q-52 (71.8-78%) and human VH4 (67.4 69.8%) gene families, which belong to mammalian group, I, clan, II, VH genes. The CDR3 length of rearranged bovine VDJ genes is characteristically long (15-23 amino acids). The bovine JH gene segments were most homologous to human JH4 (82.1 87.2%) and JH5 (84.6-89.7%) genes, suggesting the existence of at least two JH gene segments. An analysis of CDRs provides evidence that somatic hypermutations contribute significantly to the generation of antibody diversity in cattle. Southern blot analysis of BamH I, EcoR I and Hind III digested genomic DNA from four cattle breeds (Holstein, Jersey, Hereford and Charolais) revealed three RFLP patterns; the genomic complexity of Bov VH1 ranged between 13 and 15 genes. These observations provide evidence for polymorphism at the bovine Ig-VH locus, similar to that seen in mice and humans. PMID- 9393968 TI - Genomic organization of the TcR beta-chain diversity (Dbeta) and joining (Jbeta) segments in the rainbow trout: presence of many repeated sequences. AB - This work describes a 5.5 kb genomic sequence of the rainbow trout T-cell receptor beta-chain locus. It includes, from 5' to 3', a Dbeta gene, 10 Jbeta genes and the 5'-end of the first Cbeta exon. The trout Dbeta-Jbeta-Cbeta locus is about the same size as the mouse, rat and human homologous loci, but it is less compact and contains 10 Jbeta segments instead of the 6-7 found in mammals. The trout Dbeta coding sequence is identical to those of the mouse, rat and human Dbeta, and the Dbeta recombination signal sequences (RSS) are also very well conserved. Each trout Jbeta segment is flanked in 5' by a 7-mer RSS, which matches with the canonical conserved 7-mer sequences of all RSS. However, 6 of the 10 Jbeta segments have no characteristic 9-mer RSS, although at least some of them are well expressed (Jbeta1 and Jbeta2). The Jbeta region of the trout TcRbeta locus contains numerous micro/minisatellite repeated DNA sequences; some of these repeats contain heptamer RSS-like sequences that could interfere with Jbeta expression. Knowledge of the germline boundaries of the trout Dbeta and Jbeta ends makes it possible to evaluate precisely the exonuclease activity and N nucleotide addition at the Dbeta-Jbeta junctions of the rearranged TcRbeta chain genes. Many (40%) of the Dbeta-Jbeta junctions in the adult trout have no N nucleotides, compared to 26.4% in adult mice, and 37% of the adult trout TcRbeta transcripts are out of frame. Thus, there may be major differences in the T-cell developmental kinetics and selection in fish and mammals. PMID- 9393969 TI - Distinct tissue and cellular distribution of two major isoforms of calcineurin. AB - The protein phosphatase calcineurin is known to be an essential intracellular signal transducer involved in the TCR-mediated signal transduction pathway and is the common target of the immunosuppressive drugs cyclosporin A (CsA) and FK506. The catalytic subunit of calcineurin exists in multiple isoforms, but their functional differences are not known. It has been assumed that the alpha isoform of calcineurin is the relevant isoform mediating TCR signaling. Recently, calcineurin alpha was knocked out in mice, but no defect in the TCR-mediated IL-2 production was observed, suggesting that another isoform of calcineurin mediates the TCR signal transduction pathway. We have generated specific polyclonal antibodies against the alpha and the beta2 isoforms of calcineurin and examined their distribution in murine tissues and immune cells by immunohistochemical staining and Western blot analysis. We found that the beta2 isoform of calcineurin is predominant in T and B lymphocytes as well as in thymus compared to the alpha isoform, suggesting that the beta2 isoform may play a key role in TCR signaling. Furthermore, we observed that the two isoforms exhibit distinct expression patterns in both kidney and thymus, indicating that the two isoforms of calcineurin have distinct cellular functions. Together, these findings raise the possibility that the nephrotoxicity associated with CsA and FK506 can be reduced by designing novel inhibitors of calcineurin that target specific isoforms of the enzyme. PMID- 9393970 TI - Probing the structure of C1 with an anti-C1s monoclonal antibody: the possible existence of two forms of C1 in solution. AB - Anti-human C1s monoclonal antibody H1532, a mouse gamma-1-immunoglobulin elicited by a C1r2C1s2 immunogen, appeared to bind to the beta-domain of C1s by electron microscopy. In agreement with this observation, Western blotting demonstrated good binding to unreduced C1s, but no binding to the alpha or gamma-B domains. When added to solutions of the C1r2C1s2 tetramer, HI532 converted the 8.7 S tetramer into an 18 S complex, which was seen by electron microscopy to be a dimer of parallel C1s x C1r x C1r x C1s molecules cross-linked by two bivalent monoclonal antibodies. If increasing amounts of HI532 were added to C1r2C1s2 followed by addition of equivalent C1q, there was a progressive loss of hemolytic activity, which became zero when two equivalents of antibody HI532 were added. When two equivalents of HI532 were added to serum or C1 reconstituted overnight from purified subcomponents, there was an immediate loss of approximately 50% of the hemolytic activity; thereafter, activity decayed slowly and even after 24 hr, 10-30% of the activity remained. The rapid loss of only 50% of the activity would be readily explained by the existence of two conformations of C1, one of which was rapidly disassembled by antibody, and the other was resistant to disassembly. These two conformations may correspond to two previously proposed structures for the C1 complex. PMID- 9393971 TI - Wnt-10b directs hypermorphic development and transformation in mammary glands of male and female mice. AB - Wnt-10b is expressed during the formation of the mammary rudiment in mouse embryos and its expression continues through puberty when the mammary ductal pattern is established under control of ovarian steroids. Recently, viral activation of the Wnt-10b locus has linked its overexpression to mammary tumor formation, suggesting a role for Wnt-10b in patterning and growth-regulation of the mammary gland. To test this notion, we created lines of transgenic mice that express elevated levels of Wnt-10b under the control of the MMTV promoter/enhancer. Overexpression of this gene resulted in profound developmental alterations in the mammary gland, including expanded glandular development and the precocious appearance of alveoli in virgin females. Moreover, transgenic male mice also exhibited dramatic mammary development involving highly branched mammary ducts and gynecomastia. Aberrant expression of Wnt-10b in the mammary rudiments of males evidently bypasses the normal requirement for ovarian hormonal control in stimulating mammary ductal growth and the repressive effects of androgens. In addition to these developmental effects, transgenic mice of both sexes were highly susceptible to the development of mammary adenocarcinomas. Such tumors arose in a solitary manner indicating that Wnt-10b is a proto-oncogene which provides a necessary, but insufficient signal for oncogenesis. Relevant to this, there was no evidence of amplified expression of FGF mRNAs in these tumors though the Fgf's are a class of genes often implicated as collaborators in Wnt mediated tumor formation. Indeed, co-expression of MMTV-Wnt-10b and MMTV-FGF 3/int-2 resulted in sterile offspring with highly disorganized mammary epithelium, demonstrating a potent interaction between their respective developmental pathways. These results suggest that Wnt-10b, or other Wnt genes expressed early in mammary development, play a role in regulating sexual dimorphism and show potent transforming activity when overexpressed. PMID- 9393972 TI - A novel brain-specific p53-target gene, BAI1, containing thrombospondin type 1 repeats inhibits experimental angiogenesis. AB - The genetic alteration of p53 is associated with neovascularization during progression of glioma to its more malignant form, glioblastoma. Hence, one or more of the genes transactivated by p53 is likely to function as an angiogenesis inhibitors. We isolated a novel p53-inducible gene that encodes a 1584-amino-acid product containing five thrombospondin type 1 (TSP-type 1) repeats and is specifically expressed in the brain. A recombinant protein corresponding to the TSP-type 1 repeats of this gene product inhibited in vivo neovascularization induced by bFGF in the rat cornea. The expression of this gene, designated BAI1 (brain-specific angiogenesis inhibitor 1) was absent or significantly reduced in eight of nine glioblastoma cell lines, suggesting BAI1 plays a significant role in angiogenesis inhibition, as a mediator of p53. PMID- 9393973 TI - Dominant-negative abrogation of connexin-mediated cell growth control by mutant connexin genes. AB - Connexin genes exert negative growth control when transfected into various types of tumor cell lines. We previously demonstrated that connexin 26 (Cx26) suppresses in vitro and in vivo growth of HeLa cells. In this study, we have examined whether certain Cx26 mutants can abrogate cell growth control and the gap junctional intercellular communication (GJIC) capacity of such Cx26 transfected HeLa cells. For this purpose, we transfected three mutated Cx26 genes (C60F, P87L and R143W) into HeLa cells already containing the wild-type Cx26 gene, which are GJIC-competent and non-tumorigenic. Transfection of P87L and R143W mutants enhanced the tumorigenicity of the HeLa Cx26 cells in nude mice without any change in GJIC capacity. On the other hand, transfection of the C60F mutant reduced the GJIC capacity of HeLa Cx26 cells without affecting their growth in vivo. Immunostaining studies demonstrated that the Cx26 proteins were localized mainly at cell-cell contact areas in the HeLa Cx26 cells both before and after transfection of mutated Cx26 genes. These results suggest that certain mutant Cx26 proteins exert a dominant-negative effect on Cx26-regulated growth of HeLa cells and that such effects may be independent of the effect on GJIC ability. It is proposed that wild-type and mutant Cx26 proteins produce heteromeric connexons and that such heteromeric connexons may exert different effects on growth control from those of homomeric connexons. PMID- 9393974 TI - Suppression of anchorage-independent growth and matrigel invasion and delayed tumor formation by elevated expression of fibulin-1D in human fibrosarcoma derived cell lines. AB - Using differential display, we identified an mRNA that is markedly down-regulated in cell line 6A/SB1, derived from a fibrosarcoma formed in an athymic mouse following injection of carcinogen-transformed MSU-1.1 cells. The nontumorigenic parental cell strain, MSU-1.1, expresses high levels of this mRNA. Sequencing of the corresponding cDNA fragment revealed that it corresponded to an expressed sequence tag, which ultimately led to its identification as the fibulin-1D gene. Fibulin-1 is a cysteine-rich, calcium-binding extracellular matrix and plasma protein, which has four isoforms, A-D, derived from alternative splicing. Northern and Western blotting analysis of 16 cell lines established from tumors formed in athymic mice by MSU-1.1-derived cell strains independently transformed in culture showed that 44% exhibited low level or lack of expression of fibulin 1D mRNA and protein. In a similar analysis of 15 malignant cell lines derived from patients, 80% showed low level or no expression. To study the role of fibulin-1D in transformation, we transfected 6A/SB1 cells and a human fibrosarcoma-derived cell line (SHAC) with a fibulin-1D cDNA expression construct. Transfectants displaying high levels of fibulin-1D were isolated and characterized. Elevated expression of fibulin-1D led to reduced ability to form colonies in soft agar and reduced invasive potential as tested in a matrigel in vitro invasion assay. Furthermore, expression of fibulin-1D resulted in a markedly extended latency in tumor formation in athymic mice. These results indicate that low expression of fibulin-1D plays a role in tumor formation and invasion. PMID- 9393975 TI - p38 MAP kinase activation by vascular endothelial growth factor mediates actin reorganization and cell migration in human endothelial cells. AB - Vascular endothelial growth factor (VEGF) is a potent chemotactic agent for endothelial cells. Yet the signalling pathways that modulate the motogenic effects of VEGF in vascular endothelial cells are still ill defined. In the present study, we found in primary cultures of human umbilical vein endothelial cells (HUVEC) that VEGF increased cell migration and induced a marked reorganization of the microfilament network that was characterized by the formation of stress fibers and the recruitment of vinculin to focal adhesions. VEGF also stimulated the mitogen activated protein (MAP) kinases ERK (extracellular signal-regulated kinase) and p38 (stress activated protein kinase 2), but not SAPK1/JNK (stress activated protein kinase-1/c-Jun NH2-terminal kinase). Activation of p38 resulted in activation of MAP kinase activated protein kinase-2/3 and phosphorylation of the F-actin polymerization modulator, heat shock protein 27 (HSP27). Inhibiting the VEGF-induced activation of ERK with PD098059 did not influence actin organization or cell migration but totally inhibited the VEGF-induced incorporation of thymidine into DNA. Inhibition of p38 activity by the specific inhibitor SB203580 led to an inhibition of HSP27 phosphorylation, actin reorganization and cell migration. The results indicate that the p38 pathway conveys the VEGF signal to microfilaments inducing rearrangements of the actin cytoskeleton that regulate cell migration. By modulating cell migration, p38 may thus be an important regulator of angiogenesis. PMID- 9393976 TI - Inactivation of the small GTPase Rho disrupts cellular attachment and induces adhesion-dependent and adhesion-independent apoptosis. AB - Rho small GTPases regulate a variety of cellular signaling pathways involved in cell growth and transformation. In this study, we examined potential roles for Rho in adhesion-dependent and -independent pathways regulating apoptosis. Rho GTPases are specifically inactivated by exoenzyme C3 (C3) of Clostridium botulinum. Using a novel Sindbis virus-based gene expression system, we created a double subgenomic recombinant (dsSIN:C3) capable of expressing active C3 in intact cells. Infection of L929 fibroblasts with dsSIN:C3 caused essentially complete ADP-ribosylation of intracellular Rho within 1 h. dsSIN:C3-infected cells also became rounded within 1-2 h and detached by 5 h post-infection. Infection of L929 in suspension with dsSIN:C3 disrupted the ability for normal cellular attachment and spreading. Infection of primary cell explants of chicken embryo fibroblasts (CEF) and rat aortic smooth muscle cells (RSM) with dsSIN:C3 caused cytoskeletal effects similar to those seen in L929. We also observed that C3 markedly decreased the basal phosphorylation state of focal adhesion kinase (FAK). Most intriguingly, we found that dsSIN-based expression of C3 or loss of function mutants of Rho could each induce apoptosis and, in RSM, this effect was observed to be adhesion-independent. Rho GTPases, therefore, appear to regulate signal pathways that are required for cell survival and growth that are separate from, but likely overlap with, Rho-dependent pathways involved in cellular adhesion. PMID- 9393977 TI - A functional analysis of p53 during early development of Xenopus laevis. AB - p53 is a nuclear protein that acts like a tumor suppressor and is involved in regulation of cellular growth. In Xenopus, the p53 protein is highly expressed during oogenesis and is strictly cytoplasmic in the oocyte. We have analysed its participation in DNA replication and transcription during early development, using the egg and oocyte as model-systems. The injection of sperm nuclei into Xenopus eggs is followed by DNA replication and mitotic events. We show that the endogenous p53 enters the nuclei and moves through a series of discrete sub nuclear loci whose distribution is S-phase specific. A specific peripheral nuclear localization of p53 is observed before entry into S-phase, followed by an internal localization which is strictly dependent on ongoing DNA synthesis. At no stage in the cell cycle, however, did we observe any co-localization with RPA or PCNA, which were used as initiation or elongation markers for DNA replication. We also show that injection into the nucleus of the oocyte of small amounts of either Xenopus or human p53 - less than 10% of the cytoplasmic storage - is sufficient to block RNA polymerase II-dependent transcription from a coinjected TATA-box-containing reporter plasmid. Transcription is rescued by microinjection of the TATA-box binding protein (TBP), suggesting that nuclear exclusion of p53 during oogenesis may be necessary for transcription of maternal genes. These characteristics are discussed in relation to the regulation of nuclear activities during early embryogenesis. PMID- 9393978 TI - Tumor suppressor protein p53 binds preferentially to supercoiled DNA. AB - Wild type human tumor suppressor protein p53 (expressed in insect cells) binds strongly to negatively supercoiled (sc) plasmid DNA at a native superhelix density, as evidenced by electrophoretic retardation of scDNA in agarose gels and imaging by scanning force microscopy (SFM). The binding occurs both in the presence and absence of the p53 consensus sequence. At relatively low p53/DNA ratios, binding of p53 to scDNA results in the appearance of several retarded DNA bands on the gels, similar to a conventional topoisomer ladder generated enzymatically. However, after removal of p53 by deproteination, the original mobility of the scDNA is recovered, indicating that the reduction of torsional stress accompanying p53 binding does not reflect changes in linking number. In DNA samples partially relaxed by topoisomerase I p53 binds preferentially to the scDNA molecules with the largest negative superhelix density. SFM imaging of the p53/scDNA complex reveals a partial or total relaxation of the compact scDNA, the degree of which increases with the number of bound p53 molecules. Competition assays with linear DNA reveal a preference of p53 for scDNA. In addition, scDNA induces dissociation of p53 from a preformed complex with a DNA fragment (474 bp) containing the consensus sequence. We conclude that the affinity of p53 for negatively supercoiled DNA is greater than that for the consensus sequence in linear fragments. However, thermally denatured linearized plasmid DNA is efficient in competing for the binding of p53 to scDNA, although the first retarded band (presumed to contain one bound p53 molecule) is retained in the case of the plasmid containing the consensus sequence. Thus, it appears that interactions involving both the core domain and the C-terminal domain regulate the binding of p53 to scDNA. The above results are not restricted to human p53; the wild type rat p53 protein also results in the retardation of scDNA on agarose gels. The biological implications of the novel DNA binding activities of p53 are discussed. PMID- 9393979 TI - Mouse fibroblast growth factor 10: cDNA cloning, protein characterization, and regulation of mRNA expression. AB - Fibroblast growth factor 7 (FGF-7) or keratinocyte growth factor (KGF), is a potent and specific mitogen for epithelial cells. We have recently identified a novel human FGF-7 homologue, named FGF-10. To study the expression of this new FGF family member and its regulation in wound repair, we cloned the mouse FGF-10 (mFGF-10) cDNA. The encoded protein is 92% identical to human FGF-10 and 91% identical to rat FGF-10. When expressed in mammalian 293 cells, the mFGF-10 protein was glycosylated but remained cell- or extracellular matrix-associated. Upon addition of heparin, mFGF-10 protein was released into the media. mRNA encoding mFGF-10 was relatively abundant in lung, skin, brain and heart. In the skin, both FGF-7 and mFGF-10 were expressed in the dermal, but not the epidermal compartment. In contrast to FGF-7, mFGF-10 expression was not induced during cutaneous wound repair. In cultured fibroblasts, expression of mFGF-10 was strongly repressed by transforming growth factor beta and tumor necrosis factor alpha, whereas epidermal growth factor and interleukin-1beta had no effect. These results demonstrate a differential regulation of mFGF-10 and FGF-7 expression in vitro and during the wound healing process. PMID- 9393980 TI - Reduced ability of transforming growth factor-alpha to induce EGF receptor heterodimerization and downregulation suggests a mechanism of oncogenic synergy with ErbB2. AB - The epidermal growth factor receptor (EGFR) is activated by a variety of ligands including EGF and transforming growth factor-alpha (TGFalpha), whereas no ligand for the homologous ErbB2 oncoprotein has yet been identified. Here we use both an ErbB2 phosphoantibody (aPY1222) and an activation-specific EGFR antibody to show that low concentrations of EGF induce more efficient tyrosine phosphorylation of ErbB2 in A431 cells than does equimolar TGFalpha, while EGFR is more potently activated by TGFalpha. Co-precipitation studies confirm that heterodimerization of activated EGFR and transphosphorylated ErbB2 is readily induced by EGF but not TGFalpha. EGFR downregulation is also more efficiently induced by EGF, suggesting that ligand-dependent modification of ErbB2 may be required to terminate EGFR signalling in cells expressing both receptor types. These findings indicate that EGF and TGFalpha differ in their abilities to induce tyrosine phosphorylation and heterodimerization of ErbB2, and raise the possibility that ErbB2 exerts its oncogenic effect in part by impairing TGFalpha-dependent EGFR downregulation. PMID- 9393981 TI - Among genes involved in the RB dependent cell cycle regulatory cascade, the p16 tumor suppressor gene is frequently lost in the Ewing family of tumors. AB - The pRB cell cycle regulatory cascade is frequently perturbed in neoplasia by overexpression of a component of the pRB-phosphorylating cyclin D1/CDK4 complex or by inactivation of pRB or the CDK4 inhibitors p16 and p15. We investigated the status and expression of p16, p15, CCND1, CDK4 and RB genes in the Ewing family of tumors. P16 loss was observed in 8 of 27 tumors (30%) and in 12 of 23 (52%) tumor cell lines from unrelated patients. There were no discrepancies in the p16 status between primary tumors and the corresponding cell lines and between cell lines established from consecutive tumor samples. p15 was codeleted in most cases but p15 mRNA was absent also in cell lines retaining the gene. In addition, posttranscriptional p16 inactivation was observed in two cases. Although no evidence for CDK4 or CCND1 amplification was obtained, expression of these genes varied considerably in the cell lines in a case specific manner. In wild-type p16 cell lines, pRB expression was lost in one case. Our data indicate that, despite the absence of cytogenetically detectable 9p21 chromosomal aberrations, p16 deletions constitute the most frequent secondary molecular aberration in Ewing tumors so far. These results are discussed in the context of the stage of disease and the clinical outcome of the patients. The potential prognostic impact of these findings remains to be further evaluated. PMID- 9393982 TI - Fusion of splicing factor genes PSF and NonO (p54nrb) to the TFE3 gene in papillary renal cell carcinoma. AB - We demonstrate that the cytogenetically defined translocation t(X;1)(p11.2;p34) observed in papillary renal cell carcinomas results in the fusion of the splicing factor gene PSF located at 1p34 to the TFE3 helix-loop-helix transcription factor gene at Xp11.2. In addition we define an X chromosome inversion inv(X)(p11.2;q12) that results in the fusion of the NonO (p54nrb) gene to TFE3. NonO (p54nrb), the human homologue of the Drosophila gene NonAdiss which controls the male courtship song, is closely related to PSF and also believed to be involved in RNA splicing. In each case the rearrangement results in the fusion of almost the entire splicing factor protein to the TFE3 DNA-binding domain. These observations suggest the possibility of intriguing links between the processes of RNA splicing, DNA transcription and oncogenesis. PMID- 9393983 TI - A fluorescent p53GFP fusion protein facilitates its detection in mammalian cells while retaining the properties of wild-type p53. AB - Tumor progression is often characterized by the cumulative loss of crucial cell cycle control genes and the concomitant loss of genome stability. Progressed tumors are often resistant to conventional therapies. Gene-transfer of key growth regulatory genes, such as the p53 gene, is one potential approach to treating advanced tumors. To this end, we have produced high-titer retroviruses, based on the pCL vector system, which encode a chimeric protein consisting of human wild type p53 and the green fluorescent protein (wtp53GFP). The fluorescent wtp53GFP protein and the wild-type p53 protein are recognized equally by several monoclonal p53-specific antibodies, have similar half-lives and function comparably in transactivating a p53-responsive element as well as in suppressing the growth of tumor cells. Additionally, due to its fluorescent nature, wtp53GFP facilitates the direct identification of cells expressing the p53 fusion protein. Combining the features of the pCL retroviral production system with the highly visible green fluorescent protein provides a potent tool for the delivery of p53 into cells and the subsequent detection of the protein, both in vitro and in vivo. PMID- 9393984 TI - v-Abl protein tyrosine kinase (PTK) mediated suppression of apoptosis is associated with the up-regulation of Bcl-XL. AB - We demonstrated previously that the activation of v-Abl protein tyrosine kinase (PTK) in IC.DP murine pre-mast cells resulted in suppression of apoptosis after withdrawal of interleukin 3 (IL-3), that protein kinase C (PKC) translocated to the nucleus 6 h after v-Abl PTK activation and that inhibition of PKC restored apoptosis after IL-3 deprivation in the presence of v-Abl PTK activity. Here we demonstrate that v-Abl PTK activation is followed by an approximately twofold increase in mRNA level of Bcl-XL by 6 h and a corresponding increase in Bcl-XL protein level by 24 h. Bcl-xL RNA and protein decreased in IL-3 deprived cells in the absence of v-Abl PTK activity. Exposure of cells with v-Abl PTK active to the PKC inhbitor calphostin C (125 ng/ml) prevented the increase in Bcl-xL protein and resulted in apoptosis. No changes in Bax or Bcl-2 protein level were noted after IL-3 withdrawal and/or activation of v-Abl PTK. Bak was barely detectable and Bad protein level decreased in cells undergoing apoptosis. The data suggest that suppression of apoptosis by v-Abl PTK in the absence of IL-3 is associated with PKC signalling and the upregulation of Bcl-xL in IC.DP cells. PMID- 9393985 TI - Preprints and Nature. PMID- 9393986 TI - Transgenic patents a step closer in Europe. PMID- 9393987 TI - Germany tightens grip on misconduct... PMID- 9393988 TI - Female scientists wanted -- apply to UK research councils. PMID- 9393989 TI - Canadian inquiry calls for 'safety first' blood agency. PMID- 9393990 TI - European grants to face ethics scrutiny. PMID- 9393991 TI - US air base in Japan increases monitoring of local dioxin threat. PMID- 9393992 TI - No evidence of sexism in peer review. PMID- 9393993 TI - The enzyme at the end of the food chain. PMID- 9393994 TI - Do males rely on female hormones? PMID- 9393995 TI - Poor prospects for oiled birds. PMID- 9393996 TI - No 'nanofossils' in martian meteorite. PMID- 9393997 TI - TGF-beta signalling from cell membrane to nucleus through SMAD proteins. AB - The recent identification of the SMAD family of signal transducer proteins has unravelled the mechanisms by which transforming growth factor-beta (TGF-beta) signals from the cell membrane to the nucleus. Pathway-restricted SMADs are phosphorylated by specific cell-surface receptors that have serine/threonine kinase activity, then they oligomerize with the common mediator Smad4 and translocate to the nucleus where they direct transcription to effect the cell's response to TGF-beta. Inhibitory SMADs have been identified that block the activation of these pathway-restricted SMADs. PMID- 9393998 TI - A conserved RNA-binding protein that regulates sexual fates in the C. elegans hermaphrodite germ line. AB - The nematode Caenorhabditis elegans has two sexes, males and hermaphrodites. Hermaphrodites Initially produce sperm but switch to producing oocytes. This switch appears to be controlled by the 3' untranslated region of fem-3 messenger RNA. We have now identified a binding factor (FBF) which is a cytoplasmic protein that binds specifically to the regulatory region of fem-3 3'UTR and mediates the sperm/oocyte switch. The RNA-binding domain of FBF consists of a stretch of eight tandem repeats and two short flanking regions. This structural element is conserved in several proteins including Drosophila Pumilio, a regulatory protein that controls pattern formation in the fly by binding to a 3'UTR. We propose that FBF and Pumilio are members of a widespread family of sequence-specific RNA binding proteins. PMID- 9394000 TI - Radial optic flow induces vergence eye movements with ultra-short latencies. AB - An observer moving forwards through the environment experiences a radial pattern of image motion on each retina. Such patterns of optic flow are a potential source of information about the observer's rate of progress, direction of heading and time to reach objects that lie ahead. As the viewing distance changes there must be changes in the vergence angle between the two eyes so that both foveas remain aligned on the object of interest in the scene ahead. Here we show that radial optic flow can elicit appropriately directed (horizontal) vergence eye movements with ultra-short latencies (roughly 80 ms) in human subjects. Centrifugal flow, signalling forwards motion, increases the vergence angle, whereas centripetal flow decreases the vergence angle. These vergence eye movements are still evident when the observer's view of the flow pattern is restricted to the temporal hemifield of one eye, indicating that these responses do not result from anisotropies in motion processing but from a mechanism that senses the radial pattern of flow. We hypothesize that flow-induced vergence is but one of a family of rapid ocular reflexes, mediated by the medial superior temporal cortex, compensating for translational disturbance of the observer. PMID- 9393999 TI - A role for oestrogens in the male reproductive system. AB - Oestrogen is considered to be the 'female' hormone, whereas testosterone is considered the 'male' hormone. However, both hormones are present in both sexes. Thus sexual distinctions are not qualitative differences, but rather result from quantitative divergence in hormone concentrations and differential expressions of steroid hormone receptors. In males, oestrogen is present in low concentrations in blood, but can be extraordinarily high in semen, and as high as 250 pg ml(-1) in rete testis fluids, which is higher than serum oestradiol in the female. It is well known that male reproductive tissues express oestrogen receptors, but the role of oestrogen in male reproduction has remained unclear. Here we provide evidence of a physiological role for oestrogen in male reproductive organs. We show that oestrogen regulates the reabsorption of luminal fluid in the head of the epididymis. Disruption of this essential function causes sperm to enter the epididymis diluted, rather than concentrated, resulting in infertility. This finding raises further concern over the potential direct effects of environmental oestrogens on male reproduction and reported declines in human sperm counts. PMID- 9394001 TI - Defective limbic system in mice lacking the tailless gene. AB - The gene tailless is a member of the superfamily of genes that encode transcription factors of the ligand-activated nuclear receptor type, and is expressed in the invertebrate and vertebrate brain. In mice, its transcripts are restricted to the periventricular zone of the forebrain, the site of origin of neurons and glia. Here we use homologous recombination to generate mice that lack a functional tailless protein. Homozygous mutant mice are viable at birth, indicating that tailless is not required for prenatal survival; however, adult mutant mice show a reduction in the size of rhinencephalic and limbic structures, including the olfactory, infrarhinal and entorhinal cortex, amygdala and dentate gyrus. Both male and female mice are more aggressive than usual and females lack normal maternal instincts. These animals therefore enable a molecular approach to be taken towards understanding the genetic architecture and morphogenesis of the forebrain. PMID- 9394002 TI - Activation of peripheral CB1 cannabinoid receptors in haemorrhagic shock. AB - Anandamide, an endogenous cannabinoid ligand, binds to CB1 cannabinoid receptors in the brain and mimics the neurobehavioural actions of marijuana. Cannabinoids and anandamide also elicit hypotension mediated by peripheral CB1 receptors. Here we report that a selective CB1 receptor antagonist, SR141716A, elicits an increase in blood pressure in rats subjected to haemorrhagic shock, whereas similar treatment of normotensive rats or intracerebroventricular administration of the antagonist during shock do not affect blood pressure. Blood from haemorrhaged rats causes hypotension in normal rats, which can be prevented by SR141716A but not by inhibition of nitric oxide synthase in the recipient. Macrophages and platelets from haemorrhaged rats elicit CB1 receptor-mediated hypotension in normotensive recipients, and incorporate arachidonic acid or ethanolamine into a product that co-elutes with anandamide on reverse-phase high performance liquid chromatography. Also, macrophages from control rats stimulated with ionomycin or bacterial phospholipase D produce anandamide, as identified by gas chromatography and mass spectrometry. These findings indicate that activation of peripheral CB1 cannabinoid receptors contributes to haemorrhagic hypotension, and anandamide produced by macrophages may be a mediator of this effect. PMID- 9394003 TI - Leptin inhibits hypothalamic neurons by activation of ATP-sensitive potassium channels. AB - Leptin, the protein encoded by the obese (ob) gene, is secreted from adipose tissue and is thought to act in the central nervous system to regulate food intake and body weight. It has been proposed that leptin acts in the hypothalamus, the main control centre for satiety and energy expenditure. Mutations in leptin or the receptor isoform (Ob-R[L]) present in hypothalamic neurons result in profound obesity and symptoms of non-insulin-dependent diabetes. Here we show that leptin hyperpolarizes glucose-receptive hypothalamic neurons of lean Sprague-Dawley and Zucker rats, but is ineffective on neurons of obese Zucker (fa/fa) rats. This hyperpolarization is due to the activation of a potassium current, and is not easily recovered on removal of leptin, but is reversed by applying the sulphonylurea, tolbutamide. Single-channel recordings demonstrate that leptin activates an ATP-sensitive potassium (K[ATP]) channel. Our data indicate that the K(ATP) channel may function as the molecular end-point of the pathway following leptin activation of the Ob-R(L) receptor in hypothalamic neurons. PMID- 9394004 TI - RGS8 accelerates G-protein-mediated modulation of K+ currents. AB - Transmembrane signal transduction via heterotrimeric G proteins is reported to be inhibited by RGS (regulators of G-protein signalling) proteins. These RGS proteins work by increasing the GTPase activity of G protein alpha-subunits (G alpha), thereby driving G proteins into their inactive GDP-bound form. However, it is not known how RGS proteins regulate the kinetics of physiological responses that depend on G proteins. Here we report the isolation of a full-length complementary DNA encoding a neural-tissue-specific RGS protein, RGS8, and the determination of its function. We show that RGS8 binds preferentially to the alpha-subunits G(alpha)o and G(alpha)i3 and that it functions as a GTPase activating protein (GAP). When co-expressed in Xenopus oocytes with a G-protein coupled receptor and a G-protein-coupled inwardly rectifying K+ channel (GIRK1/2), RGS8 accelerated not only the turning off but also the turning on of the GIRK1/2 current upon receptor stimulation, without affecting the dose response relationship. We conclude that RGS8 accelerates the modulation of G protein-coupled channels and is not just a simple negative regulator. This property of RGS8 may be crucial for the rapid regulation of neuronal excitability upon stimulation of G-protein-coupled receptors. PMID- 9394005 TI - Pore-forming segments in voltage-gated chloride channels. AB - The ability to differentiate between ions is a property of ion channels that is crucial for their biological functions. However, the fundamental structural features that define anion selectivity and distinguish anion-permeable from cation-permeable channels are poorly understood. Voltage-gated chloride (Cl-) channels belonging to the ClC family are ubiquitous and have been predicted to play important roles in many diverse physiological and pathophysiological processes. We have identified regions of a human skeletal muscle ClC isoform that contribute to formation of its anion-selective conduction pathway. A core structural element (P1 region) of the ClC channel pore spans an accessibility barrier between the internal and external milieu, and contains an evolutionarily conserved sequence motif: GKxGPxxH. Neighbouring sequences in the third and fifth transmembrane segments also contribute to isoform-specific differences in anion selectivity. The conserved motif in the Cl- channel P1 region may constitute a 'signature' sequence for an anion-selective ion pore by analogy with the homologous GYG sequence that is essential for selectivity in voltage-gated potassium ion (K+) channel pores. PMID- 9394006 TI - Ammonia mediates communication between yeast colonies. AB - Under certain growth conditions unicellular organisms behave as highly organized multicellular structures. For example, the fruiting bodies of myxobacteria and of the slime mould Dictyostelium discoideum form structures composed of non-dividing motile cells. Although non-motile, yeasts can create organized structures, colonies in which cells communicate and act in a coordinated fashion. Colony morphologies are characteristic for different species and strains. Here we describe that, in addition to short-range intracolony cell-cell communication, yeasts exhibit long-distance signals between neighbouring colonies. The volatile alkaline compound ammonia, transmitted by yeast colonies in pulses, has been identified as a substance mediating the intercolony signal. The first alkaline pulse produced by neighbouring colonies is non-directed and is followed by acidification of the medium. The second pulse seems to be enhanced and is oriented towards the neighbour colony. Ammonia signalling results in growth inhibition of the facing parts of both colonies. This phenomenon is observed in different yeast genera. The presence of amino acids in the medium is required for ammonia production. Colonies derived from the yeast Saccharomyces cerevisiae shr3 mutant, defective in localization of amino-acid permeases, do not produce detectable amounts of ammonia and do not exhibit asymmetric growth inhibition. PMID- 9394008 TI - RagA is a functional homologue of S. cerevisiae Gtr1p involved in the Ran/Gsp1 GTPase pathway. AB - Human RagA and RagB is reported to be 52% identical to a putative GTPase of Saccharomyces cerevisiae, Gtr1p. According to the reported nucleotide sequence, we amplified human RagA and RagBs cDNAs from the human B cell cDNA library with PCR. Both cDNAs rescued a cold sensitivity of S. cerevisiae, gtr1-11. Furthermore, we introduced into the cloned human RagA cDNA, the mutation 'T21L' corresponding to the gtr1-11 mutation which has been reported to suppress not only all of rcc1-, temperature-sensitive mutants of Ran/Gsp1p GTPase GDP/GTP exchanging factor, but also rna1-1, a temperature-sensitive mutant of Ran/Gsp1p GTPase-activating protein. The resulting RagAgtr1-11 cDNA partially, but significantly, suppressed both rcc1- and rna1-1 mutations. These results indicated that RagA and RagBs are functional homologues of S. cervisiae Gtr1p. Interestingly, while wild-type human RagA and RagBs were localized within the cytoplasm, similar to S. cerevisiae Gtr1p, the mutated human RagAgtr1-11 corresponding to a dominant negative form of RagA was distributed in discrete speckles in the nucleus, being localized side by side with SC-35, a non-snRNP of the splicing complex. In contrast, a dominant positive form of RagA, Q66L was localized in the cytoplasm. Thus, RagA was suggested to shuttle between the cytoplasm and the nucleus, depending on the bound nucleotide state. PMID- 9394007 TI - Sphingolipid organization in biomembranes: what physical studies of model membranes reveal. AB - Recent cell biological studies suggest that sphingolipids and cholesterol may cluster in biomembranes to form raft-like microdomains. Such lipid domains are postulated to function as platforms involved in the lateral sorting of certain proteins during their trafficking within cells as well as during signal transduction events. Here, the physical interactions that occur between cholesterol and sphingolipids in model membrane systems are discussed within the context of microdomain formation. A model is presented in which the role of cholesterol is refined compared to earlier models. PMID- 9394009 TI - Analysis of Toxoplasma gondii stably transfected with a transmembrane variant of its major surface protein, SAG1. AB - We have genetically engineered Toxoplasma gondii so that its major surface antigen SAG1 is anchored by a human transmembrane domain (SAG1-TM) instead of its natural GPI anchor (SAG1-GPI) in order to initiate studies to address the function of this protein anchor in parasitic protozoa as well as to get insights into the functional role of SAG1. Our results show that SAG1-TM is correctly folded (at least as judged by the presence of conformationally dependent epitopes) and targeted to the surface of the parasite, indicating that the GPI anchor does not determine its localization nor overall three-dimensional structure. No significant difference was seen in any aspect of the growth of the SAG1-TM mutant. However, compared to the natural SAG1-GPI, SAG1-TM does not form strong associations with itself and/or other molecules in high molecular weight complexes suggesting that allowing such complexes to form may be one role of the GPI anchor. The in vitro half-life of SAG1-TM of extracellular parasites is significantly lower than that of SAG1-GPI suggesting a stabilizing function of the glycolipid anchor against degradation and/or membrane release. Antibodies to SAG1 are shed from SAG1-TM parasites as they invade, just as they are stripped from SAG1-GPI bearing parasites. The stripping, therefore, is unlikely to be driven by the action of lipases. PMID- 9394010 TI - A quantitative analysis of connexin-specific permeability differences of gap junctions expressed in HeLa transfectants and Xenopus oocytes. AB - Gap junctions provide direct intercellular communication by linking adjacent cells with aqueous pores permeable to molecules up to 1 kDa in molecular mass and 8-14 A in diameter. The identification of over a dozen connexins in the mammalian gap junction family has stimulated interest in the functional significance of this diversity, including the possibility of selectivity for permeants as seen in other channel classes. Here we present a quantitative comparison of channel permeabilities of different connexins expressed in both HeLa transfectants (rat Cx26, rat Cx32 and mouse Cx45) and Xenopus oocytes (rat Cx26 and rat Cx32). In HeLa cells, we examined permeability to two fluorescent molecules: Lucifer Yellow (LY: anionic, MW 457) and 4',6-diamidino-2-phenylindole, dihydrochloride (DAPI, cationic, MW 350). A comparison of the kinetics of fluorescent dye transfer showed Cx32, Cx26 and Cx45 to have progressively decreasing permeabilities to LY, but increasing permeabilities to DAPI. This pattern was inconsistent with selection based on physical size of the probe, nor could it be accounted for by the differences between clones in the electrical conductance of the monolayers. In Xenopus oocytes, where electrical and dye coupling could be assessed in the same cells, Cx32 coupled oocytes showed an estimated 6-fold greater permeability to LY than those coupled by Cx26, a comparable result to that seen in HeLa cells, where an approximately 9-fold difference was seen. The oocyte system also allowed an examination of Cx32/Cx26 heterotypic gap junction that proved to have a permeability intermediate between the two homotypic forms. Thus, independent of the expression system, it appears that connexins show differential permeabilities that cannot be predicted based on size considerations, but must depend on other features of the probe, such as charge. PMID- 9394012 TI - Targeted gene disruption reveals a role for vacuolin B in the late endocytic pathway and exocytosis. AB - Cells of Dictyostelium discoideum take up fluid by macropinocytosis. The contents of macropinosomes are acidified and digested by lysosomal enzymes. Thereafter, an endocytic marker progresses in an F-actin dependent mechanism from the acidic lysosomal phase to a neutral post-lysosomal phase. From the post-lysosomal compartment indigestible remnants are released by exocytosis. This compartment is characterised by two isoforms of vacuolin, A and B, which are encoded by different genes. Fusions of the vacuolin isoforms to the green fluorescent protein associate with the cytoplasmic side of post-lysosomal vacuoles in vivo. Vacuolin isoforms also localise to patches at the plasma membrane. Since vacuolins have no homologies to known proteins and do not contain domains of obvious function, we investigated their role by knocking out the genes separately. Although the sequences of vacuolins A and B are about 80% identical, only deletion of the vacuolin B gene results in a defect in the endocytic pathway; the vacuolin A knock-out appeared to be phenotypically normal. In vacuolin B- mutants endocytosis is normal, but the progression of fluid-phase marker from acidic to neutral pH is impaired. Furthermore, in the mutants post lysosomal vacuoles are dramatically increased in size and accumulate endocytic marker, suggesting a role for vacuolin B in targeting the vacuole for exocytosis. PMID- 9394011 TI - Localization of three human polypeptide GalNAc-transferases in HeLa cells suggests initiation of O-linked glycosylation throughout the Golgi apparatus. AB - O-glycosylation of proteins is initiated by a family of UDP-N acetylgalactosamine:polypeptide N-acetylgalactos-aminyltransferases (GalNAc-T). In this study, we have localized endogenous and epitope-tagged human GalNAc-T1, T2 and -T3 to the Golgi apparatus in HeLa cells by subcellular fractionation, immunofluorescence and immunoelectron microscopy. We show that all three GalNAc transferases are concentrated about tenfold in Golgi stacks over Golgi associated tubular-vesicular membrane structures. Surprisingly, we find that GalNAc-T1, -T2 and -T3 are present throughout the Golgi stack suggesting that initiation of O glycosylation may not be restricted to the cis Golgi, but occur at multiple sites within the Golgi apparatus. GalNAc-T1 distributes evenly across the Golgi stack whereas GalNAc-T2 and -T3 reside preferentially on the trans side and in the medial part of the Golgi stack, respectively. Moreover, we have investigated the possibility of O-glycan initiation in pre-Golgi compartments such as the ER. We could not detect endogenous polypeptide GalNAc-transferase activity in the ER of HeLa cells, neither by subcellular fractionation nor by situ glycosylation of an ER-retained form of CD8 (CD8/E19). However, upon relocation of chimeric GalNAc-T1 or -T2 to the ER, CD8/E19 is glycosylated with different efficiencies indicating that all components required for initiation of O-glycosylation are present in the ER except for polypeptide GalNAc-transferases. PMID- 9394013 TI - The role of NuMA in the interphase nucleus. AB - NuMA is an essential protein for the formation of spindle poles in mitosis. During interphase, NuMA is transported into the nucleus where it resides until prometaphase of the next mitotic cycle. We tested for a potential function of NuMA in interphase nuclei that were assembled from human sperm DNA using frog egg extract immunodepleted of NuMA. Despite the absence of NuMA, nuclei formed without visible changes of the chromatin structure, surrounded by an intact nuclear membrane containing pores and nuclear lamins. These nuclei were fully competent to import nuclear substrates and to replicate their DNA. By screening tissue sections of various organs, absence of NuMA from the nucleus was observed in a number of cell types, including sperm, granulocytes in the blood, and differentiated smooth and skeletal muscle fibers. Experiments on cultured myoblasts indicated that NuMA is degraded during muscle cell differentiation. The absence of NuMA in interphase nuclei of the tissues tested correlated with a non spherical, elongated or beaded nuclear morphology, suggesting that during interphase NuMA may act as a non-essential nucleoskeletal element. PMID- 9394014 TI - Identification of intracellular sites of superoxide production in stimulated neutrophils. AB - In this study, we show that superoxide production is carried out within intracellular compartments of human neutrophils and not at the plasma membrane following stimulation with phorbol myristate acetate. Oxidant production was not observed in unstimulated cells. Stimulated cells exhibited superoxide production in two distinct types of intracellular organelles. Initially, activity was detected in slender rod-shaped granules and in spherical or elliptical granules. The oxidant-producing granules fused directly with the plasma membrane or fused to form larger intracellular vesicles which then became associated with the plasma membrane. Longer periods of stimulation with PMA resulted in a decrease in the number of vesicles containing oxidant reaction product only, and an increase in structures containing both the oxidant-reaction product and ferritin particles; the latter was used herein as a marker for endocytosis. Thus a complex pattern of intracellular vesicular trafficking occurs in stimulated neutrophils. Alkaline phosphatase activity, a marker enzyme for a type of intracellular neutrophil granule was co-localized in the oxidant reaction-positive intracellular compartments. The time course of up-regulation of alkaline phosphatase activity to the cell surface parallelled the release of superoxide from stimulated cells. Results from this study demonstrate for the first time cytochemical and morphological evidence that superoxide is released from stimulated neutrophils through exocytosis of an oxidant-producing intracellular granule. PMID- 9394015 TI - Unidirectional movement of fluorescent microtubules on rows of dynein arms of disintegrated axonemes. AB - Tetramethylrhodamine-labelled microtubules were observed to move on rows of dynein arms of sea urchin sperm axonemes exposed by elastase-induced sliding disintegration. The microtubules moved towards the flagellar tip at a velocity of 3.1+/-2.1 microm second-1 (mean +/- s.d., n=53) in the presence of 0.1 mM ATP at 22 degrees C, but none moved towards the sperm head. We also examined the polarity of microtubule binding to axonemal doublet microtubules in the absence of ATP by using microtubules brightly labelled at their minus-ends. In 140 of 210 microtubules studied, they bound to axonemal microtubules with a parallel polarity. These results suggest that tightly packed dynein arms on the outer doublet microtubules of sperm axoneme preferentially bind microtubules to themselves with the same polarity as that of the axoneme and that they generate a force to move only these microtubules in the direction away from the sperm head. PMID- 9394016 TI - T cell syncytia induced by HIV release. T cell chemoattractants: demonstration with a newly developed single cell chemotaxis chamber. AB - A chemotaxis chamber has been developed to analyze both the velocity and the directionality of individual T cells in gradients of high molecular mass molecules over long periods of time. Employing this chamber, it is demonstrated that syncytia induced by HIV in SUP-T1 cell cultures release two T cell chemoattractants with approximate molecular masses of 30 and 120 kDa. Neither uninfected single cells nor polyethylene glycol-induced syncytia release detectable chemoattractant, suggesting that these chemoattractants are linked to HIV infection. Soluble gp120 functions as a T cell chemoattractant and the addition of anti-gp120 antibody to syncytium-conditioned medium blocks the high molecular mass chemoattractant activity but not the low molecular mass activity. The addition of anti-CD4 antibody to syncytium-conditioned medium also blocks the high molecular mass chemoattractant activity but not the low molecular mass activity. These results demonstrate that HIV-induced T cell syncytia release a low and a high molecular mass T cell chemoattractant, and suggest that the high molecular mass factor is gp120 and that it functions through the CD4 receptor. PMID- 9394017 TI - Platelet-derived growth factor (PDGF)-induced actin rearrangement is deregulated in cells expressing a mutant Y778F PDGF beta-receptor. AB - Platelet-derived growth factor-stimulated actin rearrangement and edge ruffle formation have previously been shown to be dependent on activation of phosphatidylinositol 3'-kinase, the activity of which also is important for directed migration of cells. This lipid kinase binds to phosphorylated tyrosine residues Y740 and Y751 in the kinase insert of the human platelet-derived growth factor ss-receptor. We examined the role of two other tyrosine residues in the kinase insert of this receptor, Y775 and Y778, for ligand-induced actin rearrangement. Both were shown to be phosphorylation sites; Y775 was only marginally phosphorylated in cells expressing the wild-type ss-receptor, whereas Y778 was phosphorylated at higher stoichiometry. Mutant receptors Y775F, Y778F and Y775/778F were active kinases and mediated proliferative responses when expressed in porcine aortic endothelial cells. Fluorescence staining of actin in platelet-derived growth factor-stimulated PAE cells revealed that Y778 is involved in regulation of the actin cytoskeleton since the cells contained, apart from edge ruffles and circular ruffles, a novel type of giant ruffle on the dorsal side of the cell, which consisted of irregular multilayered actin structures. Mutation at Y778 had no effect on activation of phosphatidylinositol 3'-kinase, nor on the GTPase activating protein of Ras and phospholipase C(gamma), and the extent of directed migration towards platelet-derived growth factor of these cells was not changed. We conclude that actin rearrangement is regulated in part by Y778 in the platelet-derived growth factor ss-receptor, potentially through binding of a novel signaling molecule to this site. PMID- 9394018 TI - In vivo association of lamins with nucleic acids in Drosophila melanogaster. AB - A 32P-labeling strategy was developed to study the interaction(s) in tissue culture cells between proteins and nucleic acids. Interphase and mitotic nuclear lamins were studied in Drosophila Kc cells. After bromodeoxyuridine incorporation and in vivo photo-crosslinking with 366 nm light, it was found that interphase lamins were associated with nucleic acid. Interactions with DNA as well as RNA were detected. In contrast, interaction of nucleic acids with mitotic lamin was not observed. Photo-crosslinking in the presence of antibiotics distamycin and/or chromomycin suggested that interphase lamins interacted with both A-T-rich DNA and G-C-rich DNA; interactions with G-C-rich DNA predominated. These results have implications for understanding the interphase organization of the higher eukaryotic cell nucleus as well as the transition of cells from interphase to mitosis. A model of nuclear organization, consistent with our results, is proposed. PMID- 9394019 TI - Mutational analysis of regulated exocytosis in Tetrahymena. AB - Genetic analysis of regulated exocytosis can be accomplished in ciliates, since mutants defective in stimulus-dependent secretion of dense-core vesicles can be identified. In Tetrahymena thermophila, secretion in wild-type cells can result in their encapsulation by the proteins released from vesicle cores. Cells with defects in secretion were isolated from mutagenized homozygous cells that were generated using a highly efficient method. Screening was based both on a visual assay for encapsulation, and on a novel panning step using differential centrifugation to take advantage of the selective mobility of mutants that fail to encapsulate upon stimulation. 18 mutants with defects in several ordered steps have been identified. Defects in a set of these could be localized to three stages: granule formation, transport to cell surface docking sites, and exocytosis itself. Mutants with defects in this last stage can be ordered into successive steps based on several criteria, including their responsiveness to multiple secretagogues and Ca2+ ionophores. The results of both somatic and genetic complementation on selected pairs also help to characterize the defective factors. PMID- 9394021 TI - Oral bioavailability and first-pass effects. AB - Existing experimental strategies for the in vivo evaluation of factors affecting oral bioavailability have been reviewed. Based on concepts that have evolved, an integrated set of strategies emerges that appears capable of providing estimates of the individual contributions attributable to absorption, losses in the gut lumen, and first-pass elimination in the gut wall and the liver. The only assumptions are linear pharmacokinetics and constant clearance between treatments. These methods are also suitable for the assessment of metabolite bioavailability after drug administration and the quantitative determination of sites of biotransformation and metabolite formation in vivo. PMID- 9394020 TI - Degradation of phagosomal components in late endocytic organelles. AB - Phagosomes are formed when phagocytic cells ingest particles such as bacteria, viruses or synthetic beads of different kinds. The environment within the phagosome gradually changes to generate degradative conditions. These changes require multiple interactions between the maturing phagosomes and the endocytic and the biosynthetic pathway. The phagosomes probably communicate with endocytic organelles by a transient fusion event, often referred to as the 'kiss-and-run' hypothesis. We have studied the role of endocytic organelles in the phagocytic pathway of J774 cells, a mouse macrophage cell line. We have used magnetic Dynabeads coated with 125ITC-IgG and 125ITC-OVA as phagocytic probes and were able to isolate the phagosomal fraction by means of a magnet. To separate lysosomes from other organelles in the endocytic pathway we allowed the cells to endocytose a pulse of colloidal gold particles complexed with ovalbumin. By combining this density shift technique with subcellular fractionation of a postnuclear supernatant in Percoll gradients we could isolate three endocytic fractions corresponding to early endosomes (the light Percoll fraction), late endosomes (the dense Percoll fraction) and lysosomes (the gold fraction). We observed that the proteins linked to the ingested beads are initially cleaved in the phagosomes. This cleavage is inhibited by leupeptin, a thiol-protease inhibitor, and requires an acidic environment. However, efficient communication between the phagosomes and the endocytic pathway leads to the transfer of dissociated phagocytosed peptides of different sizes to late endosomes and lysosomes for further processing. Consequently, the late endosomes and the lysosomes may be involved in the degradation of phagocytosed compounds. PMID- 9394022 TI - Effects of probenecid on brain-cerebrospinal fluid-blood distribution kinetics of E-Delta 2-valproic acid in rabbits. AB - E-Delta 2-valproic acid (E-Delta 2-VPA), a major active metabolite of VPA, has been proposed as an alternative to VPA because it is less hepatotoxic and is nonteratogenic. In rodents, VPA and E-Delta 2-VPA have a brain tissue/free plasma concentration ratio less than unity, which suggests rapid removal of the alkanoate anticonvulsants from the central nervous system. This study in rabbits employed a simultaneous iv infusion-ventriculocisternal (VC) perfusion technique to investigate the steady-state kinetics of E-Delta 2-VPA transport at the blood brain barrier, the blood-cerebrospinal fluid (CSF) barrier, and the neural cell membrane. Probenecid (PBD) was coadministered to probe the mediation of transport by organic anion transporter(s). Rabbits in the control group (N = 6) received an iv infusion of E-Delta 2-VPA to achieve a steady-state plasma concentration of 50 to 60 microg/ml. Blood and cisternal outflow of mock CSF perfusate were continuously sampled. Midway through the experiment, the VC perfusate was switched to one containing [3H]E-Delta 2-VPA. At 225 min, the rabbits were sacrificed, and each brain was removed and dissected into ten regions. Rabbits in the PBD group (N = 9) received an iv infusion and VC perfusion as in the control group as well as concomitant iv infusion of the inhibitor. The mean steady-state VC extraction ratio for [3H]E-Delta 2-VPA did not differ between the control and PBD groups (63.7 +/- 8.3% vs. 60. 6 +/- 9.6%), indicating the lack of a significant PBD-sensitive transport at the choroidal epithelium. Coadministration of PBD elevated brain concentration of cold E-Delta 2-VPA in the absence of a significant change in total or free steady-state plasma concentration. Mean E Delta 2-VPA brain tissue/free plasma concentration ratios in the various brain regions were 3.5- to 5.2-fold higher in PBD-treated animals than in the controls. Significant increases (3.0- to 4.5-fold) in the mean brain tissue/cisternal perfusate concentration ratios were also observed. Compartmental modeling of the steady-state distribution data suggested that clearance of E-Delta 2-VPA from the brain parenchyma is governed jointly by efflux transporters at the neural cell membrane and brain capillary endothelium. Moreover, PBD-induced elevation of E Delta 2-VPA tissue concentrations is attributed primarily to inhibition of E Delta 2-VPA efflux transport at the neural cell membrane, resulting in both intracellular trapping and greater tissue retention of E-Delta 2-VPA. PMID- 9394023 TI - The involvement of cytochrome P450 3A4 in the N-demethylation of L-alpha acetylmethadol (LAAM), norLAAM, and methadone. AB - The N-demethylation of LAAM, norLAAM, and methadone has been investigated in human liver microsomes and microsomes containing cDNA-expressed human P450s. Gas chromatography/mass spectrometry methods allowed detection of norLAAM and dinorLAAM formation from LAAM, dinorLAAM formation from norLAAM, and EDDP and EMDP formation from methadone. The rates of N-demethylation varied 4- to 7-fold in microsomes from four different donors with activities for LAAM and norLAAM consistently greater (5- to 14-fold) than for methadone. The N-demethylation of LAAM, norLAAM, and methadone were significantly inhibited by ketoconazole. IC50s could be determined for ketoconazole inhibition of LAAM and norLAAM N demethylation of 1.6 and 1.1 microM, respectively. The ability of ketoconazole to reduce methadone N-demethylation below 40% varied in regard to liver donor. No other P450-selective inhibitors reduced the average activities more than 43%. cDNA-expressed P450 3A4 N-demethylated LAAM, norLAAM, and methadone at greater rates than the other cDNA-expressed P450s studied (1A2, 2C9, 2D6, or 2E1). P450 3A N-demethylation of LAAM, norLAAM, and methadone exceeded the next most active P450, respectively, by at least 2.5, 9.6, and 13.4 times when expressed per milligram protein and by 18.2, 6.0, and 6.1 times when expressed per nanomole P450. These results suggest that P450 3A4 is the primary site of N-demethylation of LAAM, norLAAM, and methadone in human liver. Although other enzymes may also be capable of N-demethylating these compounds, identification of specific enzymes, except P450 3A4, has yet to be established. Knowledge of these enzymatic pathways is essential for assessment of the impact of metabolic drug-drug interactions on therapeutic success and/or adverse events. PMID- 9394024 TI - Cytochrome P450 isozymes involved in lisofylline metabolism to pentoxifylline in human liver microsomes. AB - We describe the kinetics of pentoxifylline formation from lisofylline in human liver microsomes using selective inhibitors of cytochrome P450 isozymes, correlation studies with specific isozyme activities, and cDNA-expressed human CYP1A2 and 2E1. A biphasic model fitted the data best for the formation of pentoxifylline, Km1 = 0.282 +/- 0.135 microM, Vmax1 = 0.003 +/- 0.001 nmol/min/mg protein, Km2 = 158 +/- 42.6 microM and Vmax2 =0.928 +/- 0.308 nmol/min/mg (N = 4). Pentoxifylline formation by the low Km isoform (200 microM lisofylline) required NADPH, was not inhibited by any isozyme-specific P450 inhibitor, and was inhibited only 10% and 20%, respectively, by aminobenzotriazole and N octamylamine. We concluded that the low Km enzyme was not a cytochrome P450. At 5 microM of lisofylline the CYP1A2 inhibitor, furafylline, inhibited pentoxifylline formation by 58.8%, and the nonspecific CYP2E1 inhibitor, diethyldithiocarbamate, inhibited pentoxifylline formation by 21.7%. When preincubated with furafylline plus diethyldithiocarbamate, inhibition of pentoxifylline formation was increased 71.4%. Microsomal CYP1A2 activity correlated with pentoxifylline formation (r2 = 0.870, p < 0.001). However, CYP2E1 activity did not correlate with pentoxifylline formation (r2 = 0.143, p = 0.181). Baculovirus insect cell expressed human CYP1A2 formed pentoxifylline at 0.987 nmol/min/nmol cytochrome P450 at 5 microM lisofylline. cDNA expressed CYP2E1 did not catalyze formation of pentoxifylline. Diethyldithiocarbamate inhibited pentoxifylline formation by 85.7% in cDNA expressed CYP1A2. We conclude that CYP1A2 is the high affinity enzyme catalyzing pentoxifylline formation from lisofylline. PMID- 9394025 TI - Nonspecific binding to microsomes: impact on scale-up of in vitro intrinsic clearance to hepatic clearance as assessed through examination of warfarin, imipramine, and propranolol. AB - The nonspecific, noncovalent binding of three drugs, imipramine, warfarin, and propranolol, to pooled human and animal liver microsomes has been determined using equilibrium dialysis in conditions where no cofactor (NADPH) was included in the incubation. The binding of warfarin was dependent upon both protein and drug concentration, whereas the binding of propranolol and imipramine was also dependent upon protein concentration but generally independent of drug concentration. At a microsomal protein concentration of 1.0 mg/ml and a warfarin concentration of 10 microM, the free fraction (fu(mic)) was 0.85. The corresponding values for propranolol and imipramine were 0.41 and 0.16, respectively. Thus, although all three drugs exhibit high binding in plasma (fu<0.1) the acidic drug warfarin differs from the basic drugs propranolol and imipramine in the extent to which each binds to microsomal protein. The binding of all three drugs to liver microsomes obtained from commonly studied animal species (rat, dog, and monkey) was almost identical to that observed in human. Additionally, the binding of warfarin and propranolol to microsomes obtained from insect cells used in baculovirus cytochrome P450 expression systems was similar to that exhibited in liver microsomes, when equal protein concentrations were compared. The enzyme kinetics of propranolol, imipramine, and warfarin oxidative metabolism were determined in pooled human liver microsomes, and the intrinsic clearance values obtained were used in scaling up to project human in vivo clearance. The values obtained by incorporating microsomal binding were compared with those in which this factor is ignored. The findings suggest that the parameter fu(mic) is important to obtain when attempting to relate in vitro intrinsic clearance to in vivo clearance. Also, this value is important to consider when comparing substrates with respect to enzyme specificity, since measured apparent KM values should be converted to true "free KM" values by correcting for the free fraction in the in vitro incubation. Furthermore, the extent of nonspecific binding to microsomes is likely an important parameter to consider when attempting to relate Ki values measured in vitro to observations of drug-drug interactions (or the lack thereof) in vivo. PMID- 9394026 TI - Species differences in metabolism of panomifene, an analogue of tamoxifen. AB - In vitro metabolism of panomifene (E-1,2-diphenyl-1--4-(2-(2-hydroxyethyl-amino) ethoxy)-phenyl--3,3,3- trifluoropropene), a novel antiestrogen against hormone dependent tumors, has been investigated using liver microsomes from mouse, rat, dog, and human. Hydroxylation and side chain modifications were the routes of panomifene metabolism. Microsomal biotransformation showed some qualitative similarities, but several differences were observed in the metabolic profiles of the four species tested. Seven metabolites were detected in the incubation mixtures analyzed by thin layer chromatography and autoradiography, although there was only one produced by all species that had lost the side chain. Among the side chain shortened metabolites, the compound that had lost the hydroxyethyl amino group was formed by the microsomal system of rodents, whereas the one that had lost the hydroxyethyl group was detected in the incubation mixtures with rat, dog, and human microsomes. Three metabolites (M1, M3, and M4) were produced exclusively by the dog. The structure of M3 was identified by mass spectroscopy as 4-hydroxy-panomifene. Furthermore, human liver microsomes formed a metabolite (M8) that was not detectable in the mixtures with mouse, rat, or dog microsomes. Its structure is suspected to be an oxidized form of panomifene with a double bound in the side chain. The structure of panomifene is analogous to tamoxifen, an antiestrogen currently used as a therapeutic agent against breast cancer, and there are some similar routes in their metabolism. The main difference is that the rate of tamoxifen biotransformation seems faster than that of panomifene. On the other hand, 4-hydroxy-panomifene is produced by only dog, while 4 hydroxylated derivative is one of the main metabolites of tamoxifen that has potent antiestrogenic activity and is considered to be responsible for the formation of DNA-adducts. PMID- 9394027 TI - Metabolism of clozapine by cDNA-expressed human cytochrome P450 enzymes. AB - The metabolism of clozapine was studied in vitro using cDNA-expressed human cytochrome P450 (CYP) enzymes 1A2, 3A4, 2C9, 2C19, 2D6, and 2E1. CYP1A2, 3A4, 2C9, 2C19, and 2D6 were able to N-demethylate clozapine. N-Oxide formation was exclusively catalyzed by CYP3A4. CYP2E1 did not metabolize clozapine. With regard to quantitative relationships, CYP1A2, 2C9, 2C19, and 2D6 displayed KM values ranging from 13 to 25 microM, whereas CYP3A4 had a 5-10 times higher KM value. CYP2C19 and 2D6 had the highest Vmax values (149-366 mol/hr/mol CYP). Taking into account the typical relative distribution of amounts of CYP enzymes in the liver, a simulation study suggested that at therapeutic concentrations CYP2C19 and CYP3A4 each accounted for about 35% of the metabolism. At toxic concentrations, the relative importance of CYP3A4 increased. PMID- 9394028 TI - In vitro biotransformation and identification of human cytochrome P450 isozyme dependent metabolism of tazofelone. AB - Tazofelone is a new inflammatory bowel disease agent. The biotransformation of tazofelone in human livers and the cytochrome P450 responsible for the biotransformation has been studied. Two metabolites of tazofelone were formed in vitro. A sulfoxide metabolite was identified by cochromatography with authentic standards, and a quinol metabolite of tazofelone was identified by mass spectrometry and proton NMR. Sulfoxidation was catalyzed by a single enzyme system while formation of the quinol metabolite was catalyzed by a two enzyme system. The Km and Vmax values for sulfoxidation were 12.4 microM and 0.27 nmol/min/mg protein, respectively. The high affinity Km and Vmax values for the formation of the quinol metabolite were 7.5 microM and 0.17 nmol/min/mg protein, respectively. Tazofelone was incubated at 20 microM concentration with human microsomes to determine which of the cytochrome P450 isozyme(s) is involved in the oxidation of tazofelone. A strong correlation was found between the immunoquantified concentrations of CYP3A and the rates of formation of the sulfoxide and quinol metabolites of tazofelone. Similarly, significant correlations were observed between the formation of midazolam 1'-hydroxylation and the rates of formation of both metabolites of tazofelone. Inhibition studies have indicated that triacetyloleandomycin, a CYP3A specific inhibitor, almost completely inhibited the formation of both of these tazofelone metabolites. Incubations with specific cDNA expressed microsomes indicated that the formation of both the sulfoxide and quinol metabolites was highest with CYP3A4 containing microsomes. The correlation data was confirmed by inhibition studies and cDNA expressed cytochrome P450 systems demonstrating that the biotransformation of tazofelone to its metabolites is primarily mediated by CYP3A. PMID- 9394029 TI - Glucuronidation of opioids, carboxylic acid-containing drugs, and hydroxylated xenobiotics catalyzed by expressed monkey UDP-glucuronosyltransferase 2B9 protein. AB - UDP-glucuronosyltransferase (UGT) 2B9, isolated from a cynomolgus monkey liver cDNA library, is 89% identical to human UGT2B7 in primary amino acid sequence, and the two expressed enzymes were previously shown to catalyze the glucuronidation of many common endogenous substrates. The purpose of the present study was to characterize the reactivity of expressed UGT2B9 with important therapeutic agents and other xenobiotics. UGT2B9, stably expressed in human embryonic kidney 293 cells, catalyzes the 3-O- and 6-O-glucuronidation of morphine and the 6-O-glucuronidation of codeine. A number of other morphinan (e.g. naloxone, naltrexone, and nalorphine) and oripavine (e.g. buprenorphine) derivatives are substrates for this enzyme. In general, morphinan derivatives are glucuronidated at higher rates, compared with oripavines; however, glucuronidation efficiency values (Vmax/KM) for the compounds are similar. Stably expressed UGT2B9 also catalyzes the glucuronidation of profen nonsteroidal anti inflammatory drugs, fibrate hypolipidemic agents, and straight-chain fatty acids at the carboxylic acid moiety. Monoterpenoid alcohols and propanolol are glucuronidated at aliphatic hydroxyl positions. Expressed UGT2B9 exhibits enantioselective glucuronidation for (R/S)-ibuprofen, (R/S)-propanolol, and (+)/( )-menthol. The data suggest that monkey UGT2B9 and human UGT2B7 are functionally similar. PMID- 9394030 TI - Metabolism and excretion of a new antianxiety drug candidate, CP-93,393, in cynomolgus monkeys: identification of the novel pyrimidine ring cleaved metabolites. AB - The metabolism and excretion of a new anxiolytic/antidepressant drug candidate, CP-93,393, ((7S, 9aS)-1-(2-pyrimidin-2-yl-octahydro-pyrido[1, 2-a]-pyrazin-7-yl methyl)-pyrrolidine-2,5-dione) were investigated in cynomolgus monkeys after oral administration of a single 5 mg/kg dose of 14C-CP-93,393. Urine, bile, feces, and blood samples were collected and assayed for total radioactivity, parent drug, and metabolites. Total recovery of the administered dose after 6 days was 80% with the majority recovered during the first 48 hr. An average of 69% of the total radioactivity was recovered in urine, 4% in bile, and 7% in feces. Mean Cmax and AUC(0-infinity) values for the unchanged CP-93,393 were 143.2 ng/ml and 497.7 ng.hr/ml, respectively, in the male monkeys and 17.2 ng/ml and 13.7 ng.hr/ml, respectively, in the female monkeys. HPLC analysis of urine, bile, feces, and plasma from both male and female monkeys indicated extensive metabolism of CP-93,393 to several metabolites. The identification of metabolites was achieved by chemical derivatization, beta-glucuronidase/sulfatase treatment, and by LC/MS/MS, and the quantity of each metabolite was determined by radioactivity detector. CP-93,393 undergoes metabolism by three primary pathways, aromatic hydroxylation, oxidative degradation of the pyrimidine ring, and hydrolysis of the succinimide ring followed by a variety of secondary pathways, such as oxidation, methylation, and conjugation with glucuronic acid and sulfuric acid. The major metabolites, oxidation on the pyrimidine ring to form 5-OH-CP 93,393 (M15) followed by glucuronide and sulfate conjugation (M7 and M13), accounted for 35-45% of the dose in excreta. Two metabolites (M25 and M26) were formed by further oxidation of M15 followed by methylation of the resulting catechol intermediate presumably by catechol-O-methyl transferase. A novel metabolic pathway, resulting in the cleavage of the pyrimidine ring, was also identified. The metabolites (M18, M20, and M21) observed from this pathway accounted for 8-15% of the dose. Aliphatic hydroxylation of the succinimide ring was a very minor pathway in monkey. 5-Hydroxy-CP-93,393 (M15, 37-49%), its sulfate and glucuronide conjugates (M7 and M13, approximately 34%), and the pyrimidine ring cleaved product (M18, approximately 8%) were the major metabolites in monkey plasma. The identified metabolites accounted for approximately 90, 93, 97, and 92% of the total radioactivity present in urine, bile, plasma, and feces, respectively. The major in vivo oxidative metabolites were also observed after in vitro incubations with monkey liver microsomes. PMID- 9394031 TI - Mechanism-based inactivation of human liver cytochrome P450 2A6 by 8 methoxypsoralen. AB - The P450 2A6 catalyzed 7-hydroxylation of coumarin proceeded with a mean Km of 0.40 (+/-0.13) microM and Vmax of 6.34 nmol/nmol P450/min (36-fold variation) in microsomal preparations from a panel of 12 human livers. Substrate depletion was avoided during the kinetic determinations. 8-Methoxypsoralen (8-MOP) is a potent mechanism-based inactivator of human liver P450 2A6 and reconstituted purified recombinant P450 2A6 based on the following evidence: 1) 8-MOP causes time, concentration, and NADPH-dependent loss of P450 2A6 activity that is not reversed by potassium ferricyanide or extensive dialysis, 2) loss of P450 2A6 activity is associated with a loss of spectrally observable P450, 3) addition of nucleophiles or reactive oxygen scavengers to the incubations does not prevent inactivation of P450 2A6, and 4) 8-MOP-dependent P450 2A6 inactivation is inhibited (concentration dependent) by the addition of a competitive inhibitor (pilocarpine). Inactivation is selective for P450 2A6 at low concentrations of 8 MOP (2.5 microM) after short incubation time periods (3 min) and was characterized by a KI of 0.8 and 1.9 microM in a reconstituted and microsomal system, respectively, and a kinact of 1 min-1 and 2 min-1 in a reconstituted and microsomal system, respectively. A substrate depletion partition ratio of 21 was calculated for the inactivation of recombinant P450 2A6. Potency and selectivity suggest that 8-MOP could be a useful tool in vitro for evaluating P450 2A6 activity in various enzyme preparations. PMID- 9394032 TI - Gadolinium chloride inhibition of rat hepatic microsomal epoxide hydrolase and glutathione S-transferase gene expression. AB - The effects of gadolinium chloride, a Kupffer cell toxicant, on the constitutive and inducible expression of hepatic microsomal epoxide hydrolase (mEH) and glutathione S-transferase (GST) genes were examined in rats. Northern blot analysis showed that treatment of rats with GdCl3 caused suppression of mEH and GST gene expression. mEH mRNA levels were decreased in a time-dependent manner after a single injected dose of GdCl3 (10 mg/kg, iv), resulting in 95, 55, 17, 36, and 69% of the levels in untreated animals at 6, 12, 18, 24, and 48 hr after treatment, respectively. A maximal reduction in GST Ya, Yb1/2, and Yc1 mRNA levels was also noted at 18 hr after treatment with GdCl3, followed by a gradual return to levels in untreated rats at later time points. Whereas treatment of rats with thiazole, allyl disulfide, propyl sulfide, oltipraz, or clotrimazole caused 2-13-fold increases in mEH mRNA levels at 18 hr after treatment, concomitant GdCl3 treatment caused 30-70% reductions in the increases in mEH mRNA levels. The chemical-inducible mRNA levels for GST Ya, Yb1/2, and Yc1 were also significantly inhibited by GdCl3 at 18 hr after treatment. Rats treated with GdCl3 (10 mg/kg/day, iv) for 3-5 consecutive days exhibited 40-90% decreases in mEH, GST Ya, and GST Yb1/2 mRNA levels, relative to control, whereas the Yc1 mRNA level was suppressed at early times and returned to levels in untreated animals at day 5 after treatment. The mRNA levels for mEH and GST Ya in rats treated daily with both allyl disulfide (25 mg/kg, po) and GdCl3 for 3 consecutive days were 20-30% of those in rats treated with allyl disulfide alone. Western immunoblotting showed that mEH and GST Ya protein expression was decreased at 1-3 days after consecutive daily treatment with GdCl3. Whereas treatment of rats with GdCl3 at a dose of 1 mg/kg suppressed constitutive hepatic mEH gene expression by 85% at 18 hr, rats treated with CaCl2 (10 mg/kg, iv) in combination with GdCl3 (1 mg/kg, iv) showed 45% suppression of the mEH mRNA level, compared with untreated animals. GdCl3-induced suppression was also significantly reversed for GST Ya mRNA by excessive CaCl2 administration. These results demonstrate that GdCl3 effectively inhibits constitutive and inducible mEH and GST expression, with decreases in their mRNA levels. GdCl3 suppression of detoxifying enzyme expression may be associated with its blocking of intracellular Ca2+ influx, which affects signaling pathways for the expression of the genes. PMID- 9394033 TI - In vitro and in vivo effects of the arylamine human immunodeficiency virus protease inhibitor 4R-(4alpha,5alpha,6beta, 7beta)-1-[(3-(1 imidazoylcarbamoyl)phenyl)methyl]-3-[(3-aminophenyl)m ethyl]hexahydro-5,6 dihydroxy-4,7-bis(phenylmethyl)-2H-1, 3-diazepin-2-one (SD894) on rat hepatic cytochrome P450 2B and 3A. AB - The human immunodeficiency virus-1 protease inhibitor SD894 was evaluated as an inhibitor and inducer of cytochromes P450 (CYPs) in rats. After addition of 10 microM SD894 and 2 mM NADPH to liver microsomes from dexamethasone-treated rats, a type II spectrum appeared. Within 2 min, it was replaced by a type III spectrum, with absorbance maxima at 426 and 456 nm, similar to those observed with alkylamines (SKF-525A) and arylamines (p-chloroaniline). Preincubation of microsomes from dexamethasone-treated rats with SD894 and NADPH resulted in a time-dependent inhibition of testosterone 6beta-hydroxylation (CYP 3A1/2 activity), which was decreased to 25% of controls after 30 min. Testosterone 16beta-hydroxylation (CYP 2B1/2 activity) was unaffected under these conditions. Testosterone 6beta-hydroxylation rates in liver microsomes from pregnenolone 16alpha-carbonitrile-treated rats incubated with 10 microM SD894 and NADPH, washed, and reisolated by ultracentrifugation were reduced by 71%, whereas 16beta hydroxylation was unaffected by SD894. Immunoblots of liver microsomes from rats dosed iv with SD894 or ip with TAO displayed increased CYP 2B1 and CYP 3A1 levels, respectively. Testosterone 6beta-hydroxylase activity in microsomes from TAO-treated rats was greater than controls. Preincubation of these microsomes with potassium ferricyanide produced an additional 50% increase, consistent with disruption of a metabolite-CYP complex. Microsomes from SD894-treated rats displayed a 3-fold increase in testosterone 16beta-hydroxylation. Potassium ferricyanide preincubation did not increase activity. Thus, although SD894 appears to inhibit CYP in vitro in a manner typical of other amine-containing, mechanism-based inhibitors, in vivo induction by 10 mg/kg daily doses of SD894 affects a different isozyme than does inhibition. The mechanism of induction is unknown. PMID- 9394034 TI - Human lymphocyte cytochrome P450 2E1, a putative marker for alcohol-mediated changes in hepatic chlorzoxazone activity. AB - Cytochrome P450 (CYP) 2E1 is implicated in a variety of chemically initiated hepatotoxicities, including alcoholic liver disease. These pathological conditions arise from increased production of reactive intermediates caused by elevated enzyme concentrations. Thus, the ability to detect enhanced CYP2E1 levels would aid in identifying individuals at high risk for xenobiotic-promoted liver injury. With this in mind, the present investigation assessed in vivo chlorzoxazone metabolism and compared pharmacokinetic parameters with CYP2E1 expression in blood. Twenty-two subjects were recruited and divided into two groups, control subjects and alcohol abusers, based on responses to two screening questionnaires. Those individuals with higher survey scores, i.e. those who consumed alcohol more frequently, exhibited higher rates of chlorzoxazone metabolism. Indeed, a correlation (r = 0.66, p < 0.01) was obtained when scores were compared with the pharmacokinetic parameter AUC for chlorzoxazone. Lymphocyte microsomes isolated from blood samples obtained from these same individuals were subjected to immunoblot analyses to detect CYP2E1 levels. That lymphocytes contained CYP2E1 was confirmed by reverse transcription-polymerase chain reaction and sequence analysis of the cDNA. Quantification of immunoreactive bands revealed that levels of this P450 were 2.3-fold higher in alcoholics than in control subjects. This increase in lymphocyte CYP2E1 content in alcoholic subjects coincided with a 2.1-fold increase in chlorzoxazone clearance and a 2-fold decrease in the AUC for chlorzoxazone. Importantly, a correlation (r = 0.62, p < 0.01) was observed between CYP2E1 content in lymphocytes and chlorzoxazone clearance rates. Thus, monitoring lymphocyte CYP2E1 expression may provide a substitute for estimating hepatic activity of this P450. PMID- 9394035 TI - Relative contribution of human erythrocyte aldehyde dehydrogenase to the systemic detoxification of the oxazaphosphorines. AB - Detoxification of cyclophosphamide is effected, in part, by hepatic class 1 aldehyde dehydrogenase (ALDH-1)-catalyzed oxidation of aldophosphamide, a pivotal aldehyde intermediate, to the nontoxic metabolite, carboxyphosphamide. This enzyme is found in erythrocytes as well. Detoxification of aldophosphamide may also be effected by enzymes, viz. certain aldo-keto reductases, that catalyze the reduction of aldophosphamide to alcophosphamide. Such enzymes are also found in erythrocytes. Not known at the onset of this investigation was whether the contribution of erythrocyte ALDH-1 and/or aldo-keto reductases to the overall systemic detoxification of circulating aldophosphamide is significant. Thus, NAD linked oxidation and NADPH-linked reduction of aldophosphamide catalyzed by relevant erythrocyte enzymes were quantified. ALDH-1-catalyzed oxidation of aldophosphamide (160 microM) to carboxyphosphamide occurred at a mean (+/- SD) rate of 5.0 +/- 1.4 atmol/min/rbc (red blood cell). Aldo-keto reductase-catalyzed reduction of aldophosphamide (160 microM) to alcophosphamide occurred at a much slower rate, viz. 0.3 +/- 0.2 atmol/min/rbc. Thus, at a pharmacologically relevant concentration of aldophosphamide, viz. 1 microM, estimated aggregate erythrocyte ALDH-1-catalyzed aldophosphamide oxidation, viz. 2.0 micromol/min, was only about 3% of estimated aggregate hepatic enzyme-catalyzed aldophosphamide oxidation, viz. 72 micromol/min; however, this rate is greater than the estimated flow-limited rate of aldophosphamide delivery to the liver by the blood, viz. 1.5 micromol/min. These observations/considerations suggest an important in vivo role for erythrocyte ALDH-1 in systemic aldophosphamide detoxification. Erythrocyte ALDH-1-effected oxidation of other aldehydes to their corresponding acids, e.g. retinaldehyde to retinoic acid, may also be of pharmacological and/or physiological significance since a wide variety of aldehydes are known to be substrates for ALDH-1. PMID- 9394037 TI - 9,13-Dicis-retinoic acid as an isomerization product of 9-cis-retinoic acid. PMID- 9394036 TI - Induction of rat hepatic cytochrome P450 2B subfamily by azidophenobarbital, as a possible photoaffinity probe for the putative phenobarbital receptor: comparative study with modified phenobarbitals with different functional groups. AB - To study the specific target to which phenobarbital (PB) binds, resulting in the induction of cytochrome P450, we prepared two azido-PBs (AZPBs) as photoaffinity ligands. The azido substituent was introduced at the para- or meta-position of the PB aromatic ring. In this study, we estimated the utility of these compounds by examining their inducing activities in vivo in rats. Induction was assessed by immunoblotting with anti-CYP2B1/2 antibody and measuring testosterone metabolizing activity, using hepatic microsomes. Administration of p-AZPB to rats increased hepatic CYP2B1/2 protein and testosterone 16beta-hydroxylase activity, although the effects were less than those of unmodified PB. m-AZPB showed no effect in the induction of CYP2B1/2. To assess the specificity of the effects of substituents, we compared the inducing activities of p/m-nitro-PBs, p/m-amino PBs, and p/m-hydroxy-PBs with those of AZPBs. The results showed that p-nitro-PB, m-amino-PB, and p-hydroxy-PB were also potent inducers for CYP2B1/2, with lower activity than that of unmodified PB, whereas the other three isomers had no effect. These results suggest that 1) the absence of any substituents on the aromatic ring of PB is needed for maximal inducing activity and 2) substitution at the meta-position of the PB aromatic ring tends to reduce effectiveness as an inducer more than does substitution at the para-position. Because p-amino-PB and p-acetylamino-PB, the minor and major metabolites of p-AZPB, respectively, were without effect in the induction of CYP2B1/2, the effect of p-AZPB was considered to be due to the unchanged compound itself. The present study demonstrates that, based on the weak but positive ability to induce CYP2B1/2, p-AZPB may be a useful tool for identifying the putative PB receptor. PMID- 9394038 TI - Tissue engineering: generation of differentiated artificial tissues for biomedical applications. AB - A new field in biomedical science has been established. Cell biologists, engineers, and surgeons now work within a team. Artificial connective, epithelial, or neuronal tissues are being constructed using living cells and different kinds of biomaterials. Numerous companies and laboratories are presenting dynamic developments in this field. Prognoses predict that, at the beginning of the coming century, the industry of tissue engineering will reach the importance of the present genetic technology. An enormous demand for organ and tissue transplants motivates research activities and drives the acquisition of innovative techniques and creative solutions. At the front of this development is the creation of artificial skin for severely burned patients and the generation of artificial cartilage for implantation in articular joint diseases. Future challenges are the construction of liver organoids and the development of an artificial kidney on the basis of cultured cells. In this paper we show strategies, needs, tools, and equipment for tissue engineering. The presupposition for all projects is the induction, development, and maintenance of differentiation within the tissue under in vitro conditions. As experiments in conventional culture dishes continued to fail, new cell and tissue culture methods had to be developed. Tissues are cultured under conditions as close as possible to their natural environment. To optimize adherence or embedding, cells are grown on novel tissue carriers and on individually selected biomatrices or scaffolds. The tissues are subsequently transferred into different types of containers for permanent perfusion with fresh culture medium. This guarantees constant nutrition of the developing tissue and prevents the accumulation of harmful metabolites. An organo-typical environment for epithelial cells, for example, is obtained in gradient containers, which are permanently superfused at the apical and basal sides with different media. Long term experiments result in cultured tissues in a quality thus far unreached. PMID- 9394039 TI - Differential expression of the TFF-peptides xP1 and xP4 in the gastrointestinal tract of Xenopus laevis. AB - TFF-peptides (formerly P-domain peptides, trefoil factors) represent major secretory products of the mammalian gastrointestinal tract. A molecular cloning approach revealed the existence of two TFF-peptides, xP1 and xP4, also in the stomach of Xenopus laevis. Here, the localization of these two peptides by Western blot analysis as well as immunohistochemistry is presented. xP1 is found predominantly in the surface mucous cells of the stomach, whereas xP4 is mainly localized to a specific population of goblet cells in the esophagus, to mucous neck cells of the stomach, and to closely resembling cells in antral glands. xP4 in the esophagus and in the stomach differ by their N-glycosylation patterns. Compared to mammalian TFF-peptides, xP1 obviously represents the frog homologue of human TFF1 (formerly pS2) and xP4 seems to be the amphibian equivalent of human TFF2 (formerly hSP). PMID- 9394040 TI - High-affinity glutamate transporters in the rat retina: a major role of the glial glutamate transporter GLAST-1 in transmitter clearance. AB - Glutamate is the major excitatory neurotransmitter of the mammalian retina and glutamate uptake is essential for normal transmission at glutamatergic synapses. The reverse transcriptase-polymerase chain reaction (RT-PCR) has revealed the presence of three different high-affinity glutamate transporters in the rat retina, viz. GLAST-1, GLT-1 and EAAC-1. No message has been found in the retina for EAAT-4, a transporter recently cloned from human brain. By using membrane vesicle preparations of total rat retina, we show that glutamate uptake in the retina is a high-affinity electrogenic sodium-dependent transport process driven by the transmembrane sodium ion gradient. Autoradiography of intact and dissociated rat retinae indicates that glutamate uptake by Muller glial cells dominates total retinal glutamate transport and that this uptake is strongly influenced by the activity of glutamine synthetase. RT-PCR, immunoblotting and immunohistochemistry have revealed that Muller cells express only GLAST-1. The Km for glutamate of GLAST-1 is 2.1+/-0.4 microM. This study suggests a major role for the Muller cell glutamate transporter GLAST-1 in retinal transmitter clearance. By regulating the extracellular glutamate concentration, the action of GLAST-1 in Muller cells may extend beyond the protection of neurons from excitotoxicity; we suggest a mechanism by which Muller cell glutamate transport might play an active role in shaping the time course of excitatory transmission in the retina. PMID- 9394041 TI - Estrogen receptor and progesterone receptor-immunoreactive cells are not co localized with gonadotropin-releasing hormone in the brain of the female mink (Mustela vison). AB - The distribution of gonadal steroid (estrogen, progesterone) receptors in the brain of the adult female mink was mapped by immunocytochemistry. Using a monoclonal rat antibody raised against human estrogen receptor (ER), the most dense collections of ER-immunoreactive (IR) cells were found in the preoptic/anterior hypothalamic area, the mediobasal hypothalamus (arcuate and ventromedial nuclei), and the limbic nuclei (amygdala, bed nucleus of the stria terminalis, lateral septum). Immunoreactivity was mainly observed in the cell nucleus and a marked heterogeneity of staining appeared from one region to another. A monoclonal mouse antibody raised against rabbit uterine progesterone receptor (PR) was used to identify the PR-IR cells in the preoptic/anterior hypothalamic area and the mediobasal hypothalamus (arcuate and ventromedial nuclei). This study also focused on the relationship between cells containing sex steroid receptors and gonadotropin-releasing hormone (GnRH) neurons on the same sections of the mink brain using a sequential double-staining immunocytochemistry procedure. Although preoptic and hypothalamic GnRH neurons were frequently in close proximity to perikarya containing ER or PR, they did not themselves possess receptor immunoreactivity. The present study provides neuroanatomical evidence that GnRH cells are not the major direct targets for gonadal steroids and confirms for the first time in mustelids the results previously obtained in other mammalian species. PMID- 9394042 TI - Transient axonal side branches in the developing mammalian optic nerve. AB - Optic axons were labelled with horseradish peroxidase to establish the presence of side branches and examine their distribution and morphology in the developing optic nerve of the quokka wallaby, Setonix brachyurus, the cat and rat at stages when axon numbers are at their peak. In each species, three quarters of the axons were essentially straight and lacked side branches. The remaining axons took significantly longer paths and bore side branches, mostly at points where axons undulated or changed direction. Side branches occurred at intervals of 28-43 micron, had lengths of 2-3 micron and were usually simple rather than branched. A minority (1%) of the axons crossed diagonally between fascicles and two thirds of these had more side branches (interval: 10-18 microm) on the interfascicular portion than were found on the forward-directed axons. A small number of axons (0.01%) doubled back to grow retrogradely towards the eye, these axons also bore relatively more side branches (interval: 8-22 micron), especially at points where the axons changed direction. Ultrastructural reconstruction showed that side branches resembled small axonal profiles and constituted 2% of the total axon number. It is suggested that side branches are involved in the fine-tuning of growth cone navigation. Most side branches are lost by adulthood, indicating their transient nature. The absence of retrogradely-directed axons from adults suggests that cells with such axons are removed by naturally occurring cell death. PMID- 9394043 TI - Co-localization of nitric oxide synthase I (NOS I) and NMDA receptor subunit 1 (NMDAR-1) at the neuromuscular junction in rat and mouse skeletal muscle. AB - Nitric oxide synthase I (NOS I) has been localized to the skeletal muscle sarcolemma in a variety of vertebrate species including man. It is particularly enriched at neuromuscular junctions. Recently, the N-methyl-D-aspartate (NMDA) receptor subunit 1 (NMDAR-1) has been detected in the postjunctional sarcolemma of rat diaphragm, providing a clue as to the possible source of Ca2+ ions that are necessary for NOS I activation. To address this possibility, we studied the distribution of NMDAR-1 and NOS I in mouse and rat skeletal muscles by immunohistochemistry and enzyme histochemistry. NMDAR-1 and NOS I were closely associated at neuromuscular junctions primarily of type II muscle fibers. NOS I was also present in the extrajunctional sarcolemma of this fiber type. Dystrophin, beta-dystroglycan, alpha-sarcoglycan, and spectrin were found normally expressed in both the junctional and extrajunctional sarcolemma of both fiber types. By contrast, in the muscle sarcolemma of MDX mice, dystrophin and dystrophin-associated proteins were reduced or absent. NOS I immunoreactivity was lost from the extrajunctional sarcolemma and barely detectable in the junctional sarcolemma. NOS I activity was clearly demonstrable in the junctional sarcolemma by NADPH diaphorase histochemistry, especially when the two-step method was used. NMDAR-1 was not altered. These data suggest that different mechanisms act to attach NOS I to the junctional versus extrajunctional sarcolemma. It may further be postulated that NMDA receptors are involved not only in the regulation but also sarcolemmal targeting of NOS I at neuromuscular junctions of type II fibers. The evidence that glutamate may function as a messenger molecule at vertebrate neuromuscular junction is discussed. PMID- 9394044 TI - Pituitary adenylate cyclase activating polypeptide (PACAP) in the gastrointestinal tract of the rat: distribution and effects of capsaicin or denervation. AB - The expression of pituitary adenylate cyclase activating polypeptide (PACAP) was studied in the gastrointestinal tract (GI-tract) of normal rats using radioimmunoassay, chromatography, immunocytochemistry, and in situ hybridization. PACAP-38, PACAP-27, and PACAP-related peptide were demonstrated in all parts of the GI-tract, PACAP-38 being the predominant form confirmed by chromatography. PACAP-immunoreactive nerve fibers and nerve cell bodies were found in the myenteric ganglia throughout the GI-tract. PACAP-containing nerve cell bodies were also demonstrated in the submucous ganglia of the small and large intestine. The synthesis of PACAP in intrinsic neurons was confirmed by in situ hybridization. Double immunostaining showed that PACAP is present in calcitonin gene-related peptide-containing sensory nerve fibers as well as in vasoactive intestinal polypeptide (VIP)- or VIP/gastrin-releasing peptide (GRP)-containing (intramural) nerve fibers in the upper GI-tract and in anally projecting, intrinsic VIP-and VIP/nitric oxide syntase-containing nerve cell bodies and nerve fibers in the small and large intestine. Neonatal treatment with capsaicin significantly reduced the concentration of PACAP-38 in the esophagus, stomach, and colon. Extrinsic denervation decreased the PACAP-38 concentration in the stomach, while no change was observed in the small intestine. These results indicate that PACAP- immunoreactive nerve fibers in the GI-tract originate from both intrinsic (enteric) and extrinsic (presumably sensory) sources suggesting that PACAP may have diverse gastrointestinal functions. PMID- 9394045 TI - Calbindin D28k-like immunoreactivity in the developing and regenerating circumvallate papilla of the rat. AB - The distribution of calbindin D28k (CB)-like immunoreactivity (-LI) in the circumvallate papilla (CVP) was examined during development and regeneration following bilateral crush injury to the glossopharyngeal nerve in the rat. In the adult CVP, CB-like immunoreactive (-IR) nerve fibers were observed in the subgemmal region and some penetrated into the taste buds. CB-LI was also detected in the cytoplasm of the spindle-shaped gustatory cells in the lower half of the trench epithelium, which contained numerous synaptic vesicles and bundles of intermediate filaments. These CB-IR gustatory cells made synapse-like contacts with CB-IR nerve terminals. Some CB-IR nerve terminals made contacts with the gustatory cells negative for CB-LI. At least three developmental stages were defined with regard to the developmental changes in the distribution of CB-LI: (1) Stage I (embryonic day (E) 18-postnatal day (P)5): CB-IR nerve fibers appeared in the lamina propria just beneath the newly-formed CVP at E18, but the gustatory epithelium of the CVP contained no CB-IR structures. Taste buds with taste pores appeared at P1. (2) Stage II (P5-10): thin CB-IR nerve fibers began entering the trench epithelium, but no CB-IR cells were observed. (3) Stage III (P10-adult): in addition to the intragemmal and perigemmal CB-IR nerve fibers, very few CB-IR cells appeared in the taste buds around P10, and their numbers increased progressively. The changes in the distribution of taste buds and CB-LI following glossopharyngeal nerve injury were similar to those observed during development. On post-operative day (PO) 4, the taste buds and CB-IR cells decreased markedly in number. These CB-IR cells became round in shape, and the number of CB-IR nerve fibers decreased markedly. On PO8, both taste buds and CB IR cells disappeared completely. The regenerated taste buds were first observed on PO12, increased rapidly in number by PO20, and increased slowly thereafter. CB IR nerve fibers accumulated at the subgemmal region and began penetrating into the trench wall epithelium around PO16. CB-IR cells appeared between PO20 and PO24, and their numbers increased progressively and reached the normal level on PO40. The topographical localizations of the taste buds and CB-IR cells during development and regeneration were comparable to those of normal animals. The delay of the time courses for appearance of CB-IR nerve fibers and CB-IR cells compared to the appearance of taste buds during development and regeneration suggests that CB in the gustatory epithelium may participate in the survival of the taste bud cells rather than in the induction of the taste buds. PMID- 9394046 TI - Reversibility of omeprazole-evoked changes in the ultrastructure of ECL cells in the rat stomach. AB - The ECL cells are histamine- and peptide hormone-producing endocrine cells in the rat oxyntic mucosa. They are rich in secretory vesicles and also contain microvesicles and electron-dense granules. They operate under the control of circulating gastrin. In the present study, we examined the ECL-cell ultrastructure after long term treatment with omeprazole, which is known to induce hypergastrinemia, and after withdrawal of the drug. Rats received omeprazole (400 micromol/kg per day, orally) for 16 days and were killed 1, 5, 20, or 40 days after the last dose of the drug. Oxyntic mucosal specimens were processed for electron microscopy. Electron micrographs of ECL-cell profiles were analyzed planimetrically. The ECL-cell profile area increased promptly in response to omeprazole, the secretory vesicles and granules were reduced in number and volume density, the microvesicles were unchanged in number but reduced in volume density, and vacuoles appeared. Within a week after stopping the omeprazole treatment, the numbers and volume densities of secretory vesicles and microvesicles returned to pre-stimulation values. Also, the vacuoles disappeared promptly. The ECL-cell profile area decreased below the pre-stimulation level within five days after stopping treatment, while, in contrast, the granules increased in number and volume density. Somewhat surprisingly, the cell size and the granule compartment did not return to normal until 40 days after stopping treatment. PMID- 9394047 TI - Occurrence of a terminal vascularisation after experimental myocardial infarction. AB - Physiological data indicate a residual vascularisation within ischemic myocardial regions where necrosis of most cells have been reported to occur after myocardial infarction. We therefore studied, by means of immunohistochemistry, computer assisted morphometry, and electron microscopy, the terminal vascularisation in correlation to cardiomyocytes in ten canine hearts 1 and 3 weeks after occlusion of the left anterior descending (LAD) coronary artery. In comparison to non infarcted myocardium we found the following alterations in infarcted myocardium: (1) the area density of cardiomyocytes decreased from 98% (control) to 7.9% (1 week after occlusion) and to 2.7% (3 weeks after occlusion); (2) the number of capillaries was diminished to 11.6% and to 2.6%; respectively; (3) smooth muscle alpha-actin was induced in endothelial (EC) cells of the microvessels; and (4) terminal resistance vessels increased 11-fold and 20-fold in number, respectively. Our findings confirm the necrosis of the vast majority of cardiomyocytes and capillaries within the first 3 weeks after myocardial infarction. Besides a small number of capillaries, many terminal resistance vessels, however, seem to persist in the scarring infarcted tissue. The occurrence of these microvessels is supposed to be important for the granulation tissue as well as for the control and regulation of a residual blood supply during scar formation. PMID- 9394048 TI - CD57, a marker for B-cell activation and splenic ellipsoid-associated reticular cells of the chicken. AB - We have demonstrated that the ellipsoid-associated reticular cells of chicken spleen express CD57, a marker for B-cell activation. These cells are characterised by their spindle-shaped morphology, tissue distribution and the absence of certain leucocyte-specific markers. They are phagocytotic and possess high endogenous non-specific esterase activity. Previous reports failed to detect CD57 expression on ellipsoid-associated reticular cells, probably because the tissue sections were differently treated before immunohistochemistry. CD57 is also expressed by a small number of T-cells in the spleen and the caecal tonsils. This number is highly variable between individual chickens depending on the activation state of the immune system. Moreover, CD57 is expressed by bursal lymphocytes (90% or more) but not by B-cells of the peripheral blood. More interestingly, we have been able to discriminate and quantify three B-cell populations of the secondary lymphoid organs, i.e. resting B-cells, germinal centre B-cells and plasma cells, based on their expression levels of CD57 and Bu 1 (a pan B-cell marker). Thus, CD57 should be considered as a B-cell activation marker, rather than as a marker for bursal B-cells; it is also a valuable marker for the immunohistochemical study of ellipsoid-associated reticular cells of chicken spleen. PMID- 9394049 TI - Dynamics of monocytes/macrophages and T lymphocytes in acutely rejecting rat renal allografts. AB - We examined the infiltration of acutely rejecting renal allografts (DA-->LEW) by ED1+ and ED2+ macrophages and T lymphocytes at intervals of 24 h after transplantation. Donor and recipient macrophages were differentiated by MHC class II antigen expression in double-staining experiments with ED1. Proliferation was assayed after pulse-labelling with BrdU. We subdivided allograft infiltration into three consecutive phases: 1) During phase I on days 1 to 2 after allogeneic kidney transplantation, perivascular infiltrates developed that contained numerous donor and recipient macrophages. Allograft rejection could already be diagnosed 24 h after transplantation by perivascular infiltration of T lymphocytes, whereas T cells were rarely found in isografts. 2) Phase II of allograft rejection from day 3 to 4 was characterized by massive propagation of the infiltrate. About equal numbers of interstitial donor and recipient macrophages were counted. Both macrophages and T lymphocytes proliferated in situ and macrophages outnumbered T cells until complete rejection. 3) During phase III the allograft was destroyed. Large intravascular monocytes surprisingly expressed the ED2 antigen. In the interstitium of viable graft regions, the population of recipient macrophages grew, whereas the population of donor macrophages and of T lymphocytes decreased. PMID- 9394050 TI - Expression of the mouse histone gene H1t begins at premeiotic stages of spermatogenesis. AB - The gene encoding H1t, a testicular variant of histone H1, is expressed in mammals during spermatogenesis. Northern blot and in situ hybridization has detected H1t mRNA only at the stage of pachytene spermatocytes. We have extended this analysis to more sensitive approaches and demonstrate, by RNase protection and electron-microscopic in situ hybridization, that H1t mRNA is detectable even in spermatogonia. Just a faint H1t band is seen in Western blots of nuclear protein from 9-day-old mice. This indicates that the H1t gene is expressed at premeiotic stages, albeit at a low level. In contrast to H1t mRNA, the H1t protein has not been detected in spermatogonia by electron microscopy after immunogold staining. PMID- 9394051 TI - Immunohistochemistry of matrix metalloproteinases (MMP), their substrates, and their inhibitors (TIMP) during trophoblast invasion in the human placenta. AB - The invasion of extravillous trophoblast cells into the maternal endometrium is one of the key events in human placentation. The ability of these cells to infiltrate the uterine wall and to anchor the placenta to it as well as their ability to infiltrate and to adjust utero-placental vessels to pregnancy depends, among other things, on their ability to secrete enzymes that degrade the extracellular matrix. Most of the latter enzymes belong to the family of matrix metalloproteinases. Their activity is regulated by the tissue inhibitors of matrix metalloproteinases. We have studied the distribution patterns of matrix metalloproteinases-1, -2, -3, and -9 and their inhibitors TIMP-1 and TIMP-2 as compared to the distribution of their substrates along the invasive pathway of extravillous trophoblast of 1st, 2nd, and 3rd trimester placentas by means of light microscopy on paraffin and cryostat sections as well as at the ultrastructural level (only 3rd trimester placenta). The comparison of different methods proved to be necessary, since the immunohistochemical distribution patterns of these soluble enzymes are considerably influenced by the pretreatment of tissues. All three methods revealed immunoreactivities of both, proteinases and their inhibitors, not only intracellularly in the extravillous trophoblast but also extracellularly in its surrounding matrix, the distribution patterns depending on the stage of pregnancy and on the degree of differentiation of trophoblast cells along their invasive pathway. Within the extracellular matrix, immunolocalization of matrix metalloproteinases as well as their inhibitors showed a specific relation to certain extracellular matrix molecules. PMID- 9394052 TI - Cognitive late effect factors related to decision making and risk behaviors of cancer-surviving adolescents. AB - The purpose of this correlational study was to examine factors related to cognitive late effects of treatment that may be predictors of decision making and risk behaviors for cancer-surviving adolescents. A convenience sample of 52 survivors (ages 14-19 years, disease-free for 5 years, no treatment for 2 years, and with all types of cancer except primary brain tumors) participated in this study at two regional survivor follow-up clinics. A medical record review, a semistructured interview with the teen, and intelligence testing on a separate day were used to collect data. A history of cancer therapy threatening cognitive function (defined as > or = 18 gy of radiotherapy, intrathecal or high-dose systemic methotrexate, or both) was a marginally significant predictor of poorer quality decision making in the first regression model. Poorer-quality decision making was a significant predictor of one or more risk behaviors in the second model. Younger age at initial treatment and lower cognitive ability (full-scale IQ) were not significant predictors for either of the models. There were no significant differences for the Wechsler IQ subtests related to abstract and analytic ability by cognitive threat status. Post hoc analysis indicated that lack of sensitivity to change of the Wechsler IQ measure may have affected outcomes. Abstract and/or analytic ability may be important links for decision making and risk behaviors of teen survivors, thus warranting further examination within a larger sample. Intervention to improve decision making needs to be provided for teen survivors; this may be true especially when there is a history of therapy threatening cognitive function. PMID- 9394053 TI - Predicting mammography and breast self-examination in African American women. AB - Breast cancer mortality is significantly greater in African American women than in their Caucasian counterparts. The purpose of this study was to identify variables associated with the breast cancer screening behaviors of mammography utilization and breast self-examination (BSE) in a convenience sample of low income African American women. A total of 328 African American women, living in a large midwestern metropolitan area, who were at < or = 150% of poverty level, and between the ages of 45 and 64 years were included in this study. Data were collected over a period of 18 months. Predisposing, enabling, and need variables from Anderson's theoretical framework included perceived susceptibility, benefits, barriers, confidence, knowledge, physician recommendation, demographic characteristics, and past experiences, as well as health-care and insurance information. Variables that significantly predicted mammography utilization included perceived barriers, mammography suggested by health-care professionals, recent thoughts about mammography, and a regular medical doctor. Variables that significantly predicted either frequency or proficiency of BSE included susceptibility, benefits, confidence, knowledge, barriers, and a regular physician. Implications for clinical practice include (a) recognizing predictors of screening among low-income African American women; (b) addressing culturally specific barriers, e.g., cancer fatalism, in order to increase compliance with screening; (c) establishing consistency in primary care providers; and (d) increasing confidence and knowledge through education. PMID- 9394054 TI - Enhancing breast cancer screening in the university setting. AB - Mammography, physical examination by a health care professional, and breast self examination (BSE) may increase the probability of detection of breast cancer at an early stage and thus increase long-term survivor rates. The purpose of this study was to investigate the effectiveness of supportive coaching as an intervention to enhance compliance with these breast cancer screening guidelines. The following research questions were identified: (a) what are the attitudes of women toward breast cancer screening? (b) what are the barriers to compliance identified by women in breast cancer screening? and (c) what are the effects of supportive interventions by a professional nurse and of compliance with breast cancer screening in women? A quasi-experimental design was used to study the research questions. The population chosen for the study included female employees in a state university setting. Participants were randomly assigned to one of two groups. All participants were asked to complete a prestudy questionnaire measuring attitudes and beliefs, gathering demographic and health information, and surveying breast cancer screening practices. The experimental group then received coaching and supportive interventions over the course of the academic year. The remainder of the sample served as a control group. A poststudy questionnaire was then sent to the entire sample to identify behaviors related to breast cancer screening. A variety of beliefs and attitudes were observed in the groups. No significant difference was found between the experimental and control groups on compliance with mammography and the clinical breast examination. A difference was noted on compliance with BSE by the experimental group evidencing more compliance. PMID- 9394055 TI - Stories of being a hospice nurse: a journey towards finding one's footing. AB - This article sheds light on the meaning of the lived experience of being a hospice nurse, as interpreted from in-depth interviews with 18 hospice nurses. The nurses' stories were analyzed using a phenomenologic-hermeneutic approach inspired by the philosophy of Ricoeur. Findings were synthesized into two themes: pursuing meaningful hospice care and pursuing spiritual integrity. Results indicated that it was the nurses' conceptions of "ideal" hospice practice that seemed to be the lens through which the nurses experienced and interpreted "real" practice as being either vitalizing or devitalizing. Results also indicated that being a hospice nurse means being visible as a person, in the sphere between the sacred and the profane and on the border between eternity and the finite. In narrating these experiences, the nurses used metaphors pointing towards "the sacred" and a "consciousness of fault." The tension between "ideal" and "real" hospice practices, and the nurses' vitalizing and devitalizing experiences and their use of metaphors in narrating these experiences are interpreted in the light of Bauman's two "life strategies" of deconstructing mortality and immortality, and Ricoeur's Symbolism of Evil. PMID- 9394056 TI - Relaxation to reduce nausea, vomiting, and anxiety induced by chemotherapy in Japanese patients. AB - The purpose of this study was to examine the effectiveness of progressive muscle relaxation (PMR) in reducing the nausea, vomiting, and anxiety induced by chemotherapy in Japanese patients. Subjects comprised 60 cancer chemotherapy patients who were hospitalized in a cancer center. These subjects were randomly assigned to either the experimental or control group. In addition to routine nursing care, subjects in the experimental received PMR training, while those in the control received contact with the investigator. Results from this study verified the effectiveness of PMR in reducing total scores used to measure nausea, vomiting, and retching; subscale scores of nausea; and subjective feelings of anxiety. The efficacy of PMR to reduce subscale scores of vomiting was not verified, partly due to an extremely low incidence of vomiting. PMID- 9394057 TI - Intervening with the psychosocial needs of patients and families: perceived importance and skill level. AB - Although nurses claim that psychosocial aspects of care are an important part of their practice, it is not clear that the psychosocial aspects of care is evident in clinical practice. This raises the question as to whether a gap exists between the theoretical literature and what is occurring at the bedside. This survey was designed as a means to examine (a) the importance staff nurses place on intervening with patient and family members' psychosocial needs, and (b) nurses' perceived skill level in meeting those psychosocial needs. The questionnaire, psychosocial interventions: perceived importance and skill level, was developed for this study. Questionnaires were distributed to 310 nurses, and 112 questionnaires were returned (37%). Data analyses included the use of descriptive statistics, independent t-tests, and analysis of variance. Nurses noted that each psychosocial need identified was quite important when providing care to patients with cancer and their families. However, nurses identified a moderate skill level for intervening with these aspects of care. A variety of individuals and factors were identified as promoting psychosocial care. Time was identified as the number one barrier to psychosocial care. Implications include the need for continuing education and preceptorship models that can be used to enhance nurses' skill in intervening with psychosocial needs of patients and their families. PMID- 9394058 TI - Clinical applications of genetic technologies to cancer care. PMID- 9394059 TI - Sexually transmitted diseases among teenagers in England and Wales. AB - A profile of sexually transmitted diseases (STDs) and HIV infections among teenagers in England and Wales was obtained from reports of newly diagnosed STDs among teenagers attending genitourinary medicine (GUM) clinics in 1995, laboratory reports of newly diagnosed HIV infections between 1985 when reporting began and the end of 1995, and the prevalence of HIV (unlinked anonymous programme) among teenagers attending genitourinary medicine clinics and antenatal clinics in 1994 and 1995. STD reports were analysed by sex, age group, and place of residence of patients--whether in the NHS Thames regions or elsewhere in England and Wales. High rates of STDs were reported in teenagers, particularly in girls. The incidences of gonorrhoea, chlamydia infection, and first attack genital wart infections were higher in teenage girls than in any other age group. Boys under 16 years of age had substantially higher rates of infection with all STDs in the Thames regions than elsewhere. Rates of gonorrhoea in teenagers of both sexes in the Thames regions were more than twice those in the rest of the country. Infection rates for genital herpes, and chlamydia in girls, were also higher in the Thames regions, although the geographical differences were less marked. The seroprevalence of HIV among heterosexual teenagers was very low. In contrast, 226 HIV infections among teenage boys had probably been acquired through sexual intercourse with other males. Unlinked anonymous testing revealed HIV antibody in 7.5% of routinely collected serology specimens taken from teenage homosexual or bisexual males attending GUM clinics in London. The high rates of STDs among teenage girls and all teenagers in the Thames regions make these groups a high priority for sexual health promotion, with special consideration given to homo/bisexual male teenagers. Detailed surveillance of risk factors for STDs, and further studies of teenage sexual behaviour will help to effectively target resources to improve the sexual health of teenagers in England and Wales. PMID- 9394060 TI - CD4 cell counts of 200 x 10(6)/1 or below in natural history studies and surveillance: is one enough or are two better? AB - A retrospective cohort study was performed to examine the extent and clinical significance of misclassification associated with using the current United States AIDS case defining category of an initial CD4 count < or = 200 cells x 10(6)/l (< or = 200) compared with a definition requiring two consecutive counts below this level. The main outcomes examined were the probability of subsequent CD4 counts > 200 x 10(6)/l (> 200) and progression times to AIDS and death. Of the 2025 predominantly male homosexual HIV-positive patients attending two hospital based HIV clinics with initial CD4 cell counts < or = 200, 1524 (75%) subsequently had consecutive counts < or = 200, but only half did so at the next CD4 count. Ten per cent had either no further or only non-consecutive counts < or = 200, and 15% had only one CD4 count available for analysis. The cumulative proportion of patients with a CD4 count > 200 at one year after a first count of < or = 200 was about twice (39%) that observed among the subgroup with at least two consecutive counts < or = 200 (19%). The times from the initial counts of < or = 200 to AIDS and death were also shorter by six months and two months, respectively, in the subgroup with two or more consecutive counts < or = 200. A significant proportion of patients will be prematurely classified as having a CD4 cell count < or = 200 if a single CD4 count below this level is accepted. A definition of two consecutive counts < or = 200 should be adopted in preference to a single count < or = 200 for natural history studies and clinical trials, in which precise estimates of the time to or from a defined CD4 threshold are important. In surveillance programmes, however, such an approach may be impractical because of missing or infrequent serial CD4 counts, although adjustments can be made based on these estimates of premature misclassification. PMID- 9394061 TI - Leptospirosis in the Republic of Ireland: 1985 to 1996. AB - Official government statistics and serological laboratory data provide limited information about the incidence of leptospirosis in the Republic of Ireland. The mean annual notified incidence in the Republic of Ireland from 1985 to 1996 was 1.3/million. The incidence according to hospital discharge diagnosis was higher at 4.9/million. One hundred and seventy-five serologically confirmed cases of leptospirosis were reported from 1986 to 1996, giving a mean annual incidence of 4.5/million. The true incidence of leptospirosis in the Republic of Ireland is probably higher, as hospital discharge data are incomplete and full serological testing was not always performed. Our data indicate that leptospirosis is an underestimated public health problem with only 26% of cases being notified. A national communicable disease surveillance centre in the Republic of Ireland would facilitate better monitoring and understanding of this disease. PMID- 9394062 TI - Outbreak of cryptosporidiosis associated with a swimming pool in Andover. AB - An outbreak of eight cases of cryptosporidiosis in Hampshire over a period of eight weeks in the summer of 1996 was linked to use of one swimming pool. Cryptosporidial oocysts were not isolated from samples of backwash, but the presence of enterobius ova indicated faecal contamination and a case control study including the first four primary cases suggested an association with immersion in the pool. Even in small outbreaks case control studies can provide useful supportive evidence as to the possible source of infection. PMID- 9394063 TI - Vero cytotoxin producing Escherichia coli O157. PMID- 9394064 TI - Changes in medical education: examinations under scrutiny. PMID- 9394066 TI - A survey of Irish palliative care services. AB - There has been no national policy directing the development of palliative care services in Ireland. Over the last 25 years different palliative care services have been established around the country, due largely to a strong and concerted effort on the part of voluntary groups. A study was carried out to determine the structure and process of all adult palliative care services in Ireland, to determine, where possible, the costs of providing these services and to assess the need for palliative care services in Ireland. All adult palliative care services (24 home care services, three inpatient services and one acute hospital service) in existence at the end of 1993 were circulated and 26 returns received (response rate 93 per cent). Twenty-five counties were covered by palliative care services, serving approximately three-quarters of the national population. Less than 10 per cent of patients had non-cancer diagnoses. Wide variation in staffing levels, workload, travelling, assessment of need and finance arrangements was reported. There is a need for further debate on the breadth and scope of palliative care services that should ideally be provided in Ireland, and how they should be funded in the future. PMID- 9394065 TI - The role of surgery and laser ablation in oesophageal carcinoma. AB - Between January 1990 and December 1994 oesophagectomy was carried out in 42 patients and comparison made with 38 who had palliative laser therapy. Apart from six patients referred after being unresectable at surgical exploration there were no agreed selection criteria, although the laser patients were in general older (mean 64 V 73 year) with a higher proportion of cardiorespiratory co-morbidity (14 per cent V 18 per cent). Lateral margins were involved in 14 per cent of known palliative resections with 50 per cent having positive nodes. The mean operating time was three hours and two chest drains inserted electively were removed after 3.6 days with mean drainage of 817 ml. The mean ICU stay was 5.4 days and 3 had radiological leaks; all but one settled conservatively. The 90 day mortality was 11.9 per cent for surgery and 34 per cent for laser patients. Twenty-three patients (61 per cent) required further courses of laser-therapy for benign anastomotic stenosis. Including the initial treatment of both groups 6.0 procedures per patient year were required in the laser groups compared with 1.1 for surgery. The 1, 2 and 3 year survival was 60 per cent, 31 per cent, 39 per cent for surgery compared with 24 per cent, 8 per cent, 3 per cent for laser--12 surgical patients are still alive and well at mean of 29 months (range 16-68). Surgery where possible with acceptable morbidity and mortality offers good palliation and long-term survival is possible; selection criteria for palliation only need to be defined. PMID- 9394067 TI - Irish surgeons and the Victoria Cross. PMID- 9394068 TI - Mucin-like carcinoma associated antigen (MCA) at presentation with breast cancer. AB - The usefulness of serum measurements of mucin-like carcinoma associated antigen (MCA) in 100 women at presentation with breast cancer was evaluated. Peripheral venous blood was drawn and MCA values determined by radioimmunoassay. Twenty women presenting with benign breast disease and 20 normal women served as controls. There was no difference in the MCA values between the benign breast disease group and the normal group: 4.1 +/- 0.9 units/ml versus 5.0 +/- 0.75 units/ml (mean +/- sem). The following were the MCA values for patients by stage; stage 1: 11.2 +/- 1.02, stage 2: 11.0 +/- 1.29, stage 3: 20.2 +/- 6.7, stage 4: 31 +/- 5.0. Statistical analysis of stage versus controls showed significant elevations only in stage 3 and 4 disease (p < 0.05). We conclude that MCA may be a useful serum tumour marker only in advanced breast cancer but is unreliable in detection of early breast cancer. PMID- 9394069 TI - Video-assisted thoracic surgery (VATS) for spontaneous pneumothorax. AB - Video-assisted thoracic surgery (VATS) involves using a thoracoscope with a camera chip attached to a video monitor which allows certain thoracic procedures to be performed with limited incisions. Using VATS, 170 procedures have been performed on 158 patients including 42 procedures on 39 patients with spontaneous pneumothorax. There were 24 males and 15 females with age ranging from 17 to 84 yr (mean 36.7). Indication for operation included recurrent pneumothorax in 20 (51 per cent), persistent pneumothorax in 16 (41 per cent) and bilateral pneumothorax in 3 (8 per cent). The main therapeutic strategies were apical pleurectomy, in all (42) and blebectomy/bullectomy in 38 (90 per cent). There was one hospital death (hospital mortality 2.5 per cent) in an elderly patient who developed multi organ failure post bullectomy and persistent air leak. One patient (2.5 per cent) required conversion to formal thoracotomy. Mean post operative chest tube duration was 2.7 days and mean post-operative hospital stay was 5.1 days. There has been no recurrence of pneumothorax in this series during short term follow up (mean 18 months). Our experience indicates an expanding role for video-assisted thoracic surgery in the management of patients with spontaneous pneumothorax. PMID- 9394070 TI - Dietary intakes in Ireland of a healthy elderly population. AB - The aim of this paper is to assess the adequacy of the diet of individuals over 60 yr of age, participating in the 1990 Irish National Nutritional Survey. A nationwide random sample based on the most recently updated electoral register was used. Demographic information was collected. Anthropometric measurements were taken and nutrient intake was assessed using the 7-day dietary history method. The randomly selected sample of 1213 subjects was considered to be representative of the Irish population. Of those selected, 163 individuals were over 60 yr of age, 79 of whom were male and 84 female. Mean energy intakes including alcohol for males and females were 9.55 +/- 3.09MJ and 7.07 +/- 2.39MJ respectively. The main sources of energy were bread, meat and meat products, potatoes and milk. As percentage energy, protein, fat and carbohydrate intakes were 14.90 per cent, 33.97 per cent and 48.22 per cent for men and 15.39 per cent, 34.09 per cent and 49.37 per cent for women respectively. Except for vitamin D and folate, micronutrient intakes were adequate. The body mass index (BMI [weight/height2] kg/m2) for men was 25.6 and for women 26.4. Fewer than 27.8 per cent of the males and 20.2 per cent of females take part in regular physical activity. In conclusion, the diet of a healthy elderly population in Ireland is nutritionally adequate with macronutrient intake in keeping with the recommended guidelines. Overall energy intakes are lower than those of a younger age group and may account for the lower intakes of certain micronutrients. An increase in fruit and vegetable consumption would improve vitamin and mineral intake. In order to allow for a higher energy intake an increase in physical activity is desirable. PMID- 9394071 TI - Evaluation of lifestyle, food consumption and nutrient intake patterns among Irish teenagers. AB - Lifestyle, food consumption and nutrient intake patterns from a randomly selected group of 390 secondary pupils aged between 12-18 were evaluated. Demographic information and anthropometric measurements included weight, height, and skinfold thickness were taken. Nutrient intake was assessed using the 7-day dietary history method, using a photographic atlas as an aid. Mean energy intakes for boys and girls aged 12-15 and 15-18 were 11.3MJ and 14MJ and 9.1MJ and 8.9MJ respectively. As percentage energy, protein fat and carbohydrate intakes varied little between the different age-sex groupings and were approximately 13.7-14.5, 35.4-37 and 46.8-50 per cent respectively. For boys micronutrient intake for iron and folate achieving only 83 and 78 per cent and 98 and 90 per cent of the recommended nutrient intake (R.N.I.) for ages 12-15 and 15-18 respectively. Mean dietary fibre intakes were approximately 19.6-25g/day for boys aged 12-18 and 17g/day for girls of a similar age. The main sources of energy were bread, meat and meat products, potatoes/chips, confectionery and preserves. Fruit and vegetable consumption was low for all groups. The majority of those surveyed consumed the traditional main meals. Snacking was also common practice. The snack foods consumed were generally of a high fat/high sugar content. 1.1 per cent boys and 2.6 per cent of girls aged 12-15 and 5.5 per cent and 8.2 per cent of boys and girls aged 15-18 respectively had a BMI greater than 26 indicating a risk of overweight. Greater than 68 per cent of girls and 79.5 per cent of boys surveyed participated in some form of sport. Boys were more physically active than girls and older girls less active than younger. In conclusion, changes from present day practices would be beneficial to reduce incidence of chronic disease for present day teenagers. PMID- 9394072 TI - Acute bacterial meningitis in adults: analysis of 218 episodes. AB - A retrospective study was conducted to examine the laboratory, clinical features and outcome of 206 adult acute bacterial meningitis patients (218 episodes) during the years 1985-1996. Pneumonia (8.7 per cent), head trauma (7.8 per cent) and chronic otitis media (6.0 per cent) were identified as the main predisposing factors for acute bacterial meningitis. Aetiology was described only in 61 episodes (28.0 per cent). Streptococcus pneumonia was the most commonly identified pathogen overall, causing 33 of the 218 episodes (15.2 per cent). Antibiotic treatment before admission was given to 48.4 per cent of patients. On admission, the following symptoms of meningitis were predominant: 83 per cent had neck stiffness, 81 per cent had a headache and 73 per cent had fever. Case fatality rate was 27.1 per cent (59 patients). The important factors in mortality were as follows: old age, a long duration of symptoms before admission, a lack of neck stiffness, obtunded mental state on admission, low glucose levels in first CSF, low CSF/blood glucose ratio, and abnormality in computerised tomography scanning. PMID- 9394073 TI - HIV risk behaviour in Irish intravenous drug users. AB - The aim of the study was to measure HIV prevalence and risk behaviour in 185 Irish Intravenous Drug Misusers. Information was obtained by application of a standardised WHO questionnaire covering HIV risk behaviour in the preceding 6 months. HIV serostatus was obtained by saliva/blood sample testing. One hundred and 3 (55.7 per cent) shared and 114 (61.6 per cent) lent used injecting equipment in the previous 6 months. 97 (94.2 per cent) of those who shared always cleaned the needles before use but only 48 (49.5 per cent) of these always cleaned in an efficient manner. One hundred and 14 (79.2 per cent) males and 28 (68.3 per cent) females reported heterosexual activity in the preceding 6 months. On examination sexual risk behaviour was found to be high. 50.5 per cent of males and 63 per cent of females never used condoms with regular partners. 32.6 per cent of males never used condoms with casual partners. The large majority of partners of male I.D.U'.s (both regular and casual) were non injectors. Therefore there is potential for sexual spread of HIV into the non-injecting heterosexual population. Conversely the vast majority of partners of female IDU's were injectors. This suggests that female IDU's are at higher risk of HIV infection than their male counterparts. HIV prevalence in the study group was 8.4 per cent. Implications of results for future intervention are discussed. PMID- 9394074 TI - Carbimazole induced agranulocytosis: rescue with human recombinant granulocyte colony stimulating factor. AB - We report on a 25 yr old woman with primary autoimmune hyperthyroidism who was treated with carbimazole. Forty-three days later she developed agranulocytosis (64 x 10(6)/l). With 4 days treatment with recombinant human Granulocyte Colony Stimulating factor the granulocyte count rose to normal--4,350 cells x 10(6)/l. This is considerably faster than the rate of spontaneous recovery which usually takes one to 2 weeks. PMID- 9394075 TI - Dermatitis Herpetiformis: a review of fifty-four patients. AB - A review of 54 patients with dermatitis herpetiformis presenting between 1984 1993 to The Regional Centre of Dermatology, Mater Misericordiae Hospital was undertaken. All patients had skin lesions clinically and histologically typical of dermatitis herpetiformis. Deposition of granular IgA at the dermoepidermal junction on direct immunofluorescence was present in each case. The average age of onset was 41.8 yr, patients having symptoms for an average of 1.6 yr before diagnosis. Six patients had a prior history of coeliac disease. Two patients had a family history of dermatitis herpetiformis, a father and son who were both propositi in this study. Small bowel biopsy was performed on 35 patients, 71.4 per cent of them showing evidence of villous atrophy. All patients were controlled on a gluten free diet or by dapsone or a combination of these. None of the patients experienced serious adverse effects of therapy, nor did any develop lymphoma of the small bowel with a mean follow up period of 4.2 yr (range 1-10 yr). PMID- 9394076 TI - Renal transplantation performed across a positive crossmatch: a single centre experience. AB - The importance of certain positive crossmatches (CM+) in kidney transplantation remains controversial. Fifty consecutive kidney transplants were performed across a CM+ between Jan. 1990-April 1994. In 19 cases there was an isolated B-cell CM+ (Group I), in 24 an historic T-cell IgM CM+ (Group II) and in 7 an historic T cell IgG CM+ (Group III). Comparing groups I:II:III: early acute rejection affected 32%, 42%, 57% of grafts; mean serum creatinine at 3 months was 166, 150, 229 umol/l (p < 0.05); 1 yr graft survival was 95 per cent, 96 per cent, 71 per cent (p = 0.09). In group III both graft losses were in the setting of an additional current B-cell CM+. CONCLUSIONS: Transplantation performed in either the presence of an isolated B-cell CM+ or in the presence of an historic T-cell IgM CM+ was associated with acceptable outcomes at 1 yr. An historic T-cell IgG CM+ was confirmed as a contraindication to transplantation in most circumstances, especially when coupled with a current B-cell CM+. PMID- 9394078 TI - The Melkersson Rosenthal syndrome--a differential diagnosis of facial sarcoidosis. PMID- 9394077 TI - A comparison of regular with intermittent bronchodilators in asthma patients on inhaled steroids. AB - The aim of this study was to compare the effect of regular versus intermittent (p.r.n.) bronchodilators on bronchial reactivity and asthma control in patients on concomitant inhaled corticosteroids. We studied 12 patients with asthma in a prospective, randomised, single-blind, single-dummy, three-period crossover trial comparing placebo (2 puffs t.d.s.), salbutamol (200 micrograms t.d.s.) and oxitropium bromide (200 micrograms t.d.s.) for 28 days each. Computerised spirometry and bronchial reactivity to histamine were obtained on entry and after each treatment period. Symptom scores, use of rescue bronchodilator and peak expiratory flow rates were recorded daily. There were no significant differences in bronchial hyperreactivity between the salbutamol, oxitropium and placebo treatment periods. There were no significant differences in baseline FEV1, FEF25 75%, symptom scores, use of rescue bronchodilator or morning and evening PEFR between treatment periods. Intermittent beta agonist therapy is as effective as regular therapy in terms of asthma control and bronchial hyperreactivity in patients on concomitant inhaled corticosteroid therapy. Since intermittent therapy achieves similar results with significantly lower beta agonist consumption, the data support current recommendations that beta agonists should be taken on a p.r.n. basis in asthma patients on inhaled steroids. PMID- 9394079 TI - Long-term efficacy of cyclical etidronate therapy in postmenopausal osteoporosis. AB - Sixty-two women (mean age 68.7 +/- 0.9 yr) with postmenopausal spinal osteoporosis were treated with cyclical etidronate therapy (400 mg for 2 weeks alternating with 12 weeks of 1 gm elemental calcium and 400 IU Vitamin D3) for a minimum of 2 yr. Bone mineral density (BMD) of the lumbar spine (g/cm2) increased significantly (p < 0.0001) after yr 1 (4.1 +/- 0.5 per cent) and yr 2 compared with yr 1 (2.2 +/- 0.5 per cent). The response rate was 89 per cent after yr 1 and 84 per cent after yr 2. BMD of the hip (30 patients) increased by 1.5 +/- 0.9 per cent after yr 1 and 5.5 +/- 1.1 per cent (p < 0.0001) after yr 2 when compared with baseline. The response rate was 63 per cent after yr 1 and 80 per cent after yr 2. Smaller numbers of patients continued with treatment up to 4 yr with no adverse long-term effects. PMID- 9394080 TI - Efficacy of combination therapy in non-insulin dependent diabetes mellitus. AB - Secondary failure of oral hypoglycaemic agents raises the dilemma of whether to institute therapy with insulin alone, or in combination. We reviewed our experience of combination therapy following secondary failure of oral hypoglycaemic therapy. Seventeen subjects were receiving combination therapy for 6 months or more. Such treatment was associated with a significant fall in HbA1C- from 10.7 +/- 0.38 per cent to 8.3 +/- 0.35 per cent (p < 0.01) after 6 months and remained significantly reduced at 12 months (8.7 +/- 0.34 per cent (p < 0.01)). Mean body weight, systolic and diastolic blood pressure were unchanged during treatment with adjuvant insulin therapy. Insulin therapy is a useful adjunct in the daily management of subjects with NIDDM who experience secondary failure of oral hypoglycaemic agents. PMID- 9394082 TI - Familial adenomatous polyposis registry in South Carolina. PMID- 9394081 TI - Non-admitted elderly in the accident and emergency department. AB - The aim of the study was to identify the functional disabilities and support needs of elderly people who presented but were not admitted to a Dublin Accident & Emergency (A & E) department within a 1 month period. Semi-structured interviews were conducted with 19 per cent (100/532) of the non-admitted elderly within 2 weeks of the A & E visit. Injury related complaints were apparent in 51 per cent of the patients with 3 per cent requiring hospital admission within 2 weeks of the A & E visit. Increased dependency in 1 or more Activities of Daily Living (ADL) occurred in 10 per cent while 28 per cent had increased dependency in 1 or more Instrumental Activities of Daily Living (IADL). Increased family support following discharge was received by 45 per cent of the elderly. The most commonly needed statutory service which was not provided was the home-help service. This study provides baseline data on the non-admitted elderly in one Dublin A & E department and should assist planning of future service. PMID- 9394083 TI - Percutaneous method for internal jugular venous introduction and retroclavicular placement of permanent endocardial leads. PMID- 9394084 TI - Oocyte donation program: initial experiences at Southeastern Fertility Institute. AB - From March 1994 to February 1996, 28 infertile couples participated in the oocyte donation program in 33 treatment cycles at the Southeastern Fertility Institute. Of the 31 cycles with embryo transfer, 15 cycles (48.4 percent) resulted in a clinical pregnancy with fetal heart beat by ultrasound. The spontaneous first trimester abortion rate was 3/15 (20 percent), multiple pregnancy rate 3/15 (20 percent), live birth rate 11/15 (73.3 percent) and delivery rate 12/15 (80 percent). It is recommended that oocyte donation procedure is a highly successful treatment option for women with ovarian failure or repeated unsuccessful trials of assisted reproductive technologies. PMID- 9394085 TI - The origin of hemodialysis in South Carolina. PMID- 9394086 TI - Complexity in health: race, gender, social support, and campus culture. PMID- 9394087 TI - The effect of family and social support on feelings and past acts of violence among African American college men. AB - A sample of 1,874 male undergraduates in 11 predominantly African American colleges and universities was surveyed to explore the effect of social support and family factors on feelings and past acts of violence. The health of the students' interpersonal relationships in the family during adolescence, as well as the informational and emotional support from the students' mothers and significant others, were found to be significantly associated with feelings of violence and past acts of violence. Various strategies for reducing violence among African American men, including violence-prevention programs at the college level, for families, and for the community, are discussed. Changes in public- and private-sector programs to reduce and prevent violence in American communities are called for. PMID- 9394088 TI - Health behaviors of students attending historically black colleges and universities: results from the National College Health Risk Behavior Survey. AB - The National College Health Risk Behavior Survey was administered to a convenience sample of students at 8 historically Black colleges and universities in 7 states. Analyses showed major differences in the men's and women's responses. The men were significantly more likely than the women to be current smokers. Also, they more frequently reported heavy drinking, drinking and driving in the past days 30 days, and carrying a weapon. The women were significantly more likely to view themselves as overweight, and more than one third reported they were trying to lose weight. More than one third of the students had not exercised or participated in sports activities for more than 20 minutes in the past 7 days. Because historically Black colleges and universities educate the majority of undergraduate Black college students, multidimensional programs to promote disease prevention and health promotion for Black college students must be expanded and evaluated. PMID- 9394089 TI - How sleep deprivation affects psychological variables related to college students' cognitive performance. AB - The effects of sleep deprivation on cognitive performance psychological variables related to cognitive performance were studied in 44 college students. Participants completed the Watson-Glaser Critical Thinking Appraisal after either 24 hours of sleep deprivation or approximately 8 hours of sleep. After completing the cognitive task, the participants completed 2 questionnaires, one assessing self-reported effort, concentration, and estimated performance, the other assessing off-task cognitions. As expected, sleep-deprived participants performed significantly worse than the nondeprived participants on the cognitive task. However, the sleep-deprived participants rated their concentration and effort higher than the nondeprived participants did. In addition, the sleep-deprived participants rated their estimated performance significantly higher than the nondeprived participants did. The findings indicate that college students are not aware of the extent to which sleep deprivation negatively affects their ability to complete cognitive tasks. PMID- 9394090 TI - College women's perceptions regarding resistance to sexual assault. AB - College women's perceptions about resistance to sexual assault were examined. Twenty-one percent of the 334 women surveyed stated that they had been sexually assaulted. The vast majority of participants had changed their lifestyles to prevent a sexual assault. Less than 1 woman in 5 of those surveyed had taken a self-defense class. Participants believed that resisting sexual assault by a stranger with a weapon was more likely than resisting an unarmed attacker to increase their chances of being physically harmed, raped, or murdered. Twenty-two percent of the participants said they were "very likely" to resist sexual assault by a stranger with a weapon; 52% would resist a stranger without a weapon. The findings indicate the need for an increase in the number of women taking self defense classes and a revision in women's perceptions about resisting sexual assault. PMID- 9394091 TI - Cessation related perceptions and behavior of former and current smokeless tobacco users. AB - Four hundred fourteen former and 73 current users of smokeless tobacco were questioned about their experiences in giving up smokeless tobacco. Their responses were compared with those of 463 ex-smokers to determine whether former smokeless tobacco users differed from former smokers in using specific cessation techniques. Of the smokeless tobacco users, 77% were interested in quitting, but only 7% wanted to quit "now." Seven percent of the daily users reported that a college-based health or fitness professional had advised them to quit. Former smokeless tobacco users were significantly more likely than former smokers to report that smoking cigarettes was related to their efforts to give up smokeless tobacco than former smokers were to report using smokeless tobacco as a strategy to stop smoking, Former smokeless tobacco users were also significantly more likely than former smokers to report current tobacco use. Smokeless tobacco cessation programs based on the transtheoretical approach to behavior change are recommended. PMID- 9394092 TI - Health discussions between college students and parents: results of a Delphi study. AB - College students' perspectives on health discussions with parents were examined in a survey of students from 5 US universities. Topics important for promoting students' and parents' health as well as guidelines for promoting productive discussions between college students and their parents were identified, using the Delphi technique to reach consensus. Sex, drugs, alcohol, and HIV/AIDS ranked as the most important discussion topics related to students' health; family relationships, physical fitness, and stress management ranked as most important topics dealing with parental health. Guidelines for profitable discussion were that the individual be honest, open, respectful of others' opinions, and a good listener. Parent education, health education activities that promote student parent discussion, and further research on the topic are suggested. PMID- 9394093 TI - Frank B. Cerra, MD. AB - Though the effects of managed care can be seen all across the country, the state of Minnesota has clearly been in the forefront of change. While this has presented an opportunity to be on the leading edge of health reform, it has also had a revolutionary impact on all previously held ways of doing business. A long time faculty member at the University of Minnesota Frank Cerra was named Dean of the medical school in May of 1995, a time which found the University of Minnesota Hospital in a precarious position. The day-to-day financial workings of the institution soon became his major focus and in 1997 he became the Senior Vice President for Health Sciences. In this position much of the restructuring and strategic planning of the school is now under his supervision, and he has dealt with several daunting challenges both within the school and the state. Interviewed in his office in Minnesota, Cerra candidly reflected on the power of market-based health reform the frustrations involved in turning a slow moving public institution towards the future. PMID- 9394094 TI - Ethical and regulatory challenges in a randomized control trial of adjuvant treatment for breast cancer in Vietnam. PMID- 9394095 TI - Cyclic AMP-mediated signal transduction in heart failure: molecular pathophysiology and therapeutic implications. PMID- 9394096 TI - Testosterone suppression of the HPT axis. AB - BACKGROUND: Although studies have demonstrated the suppression of normal gonadal function in the experimental setting, the specific mechanisms by which androgenic anabolic steroids impact male gonadal function remain ill defined. Following 2 consecutive weekly injections of an identically appearing testosterone cypionate (TC) placebo, subjects were randomized to a TC dose of 100 mg/wk, 250 mg/wk, or 500 mg/wk. Following the last weekly injection of active agent the subjects received 12 consecutive weeks of TC placebo injections. RESULTS: Spermatogenesis was impaired by each of the doses of TC employed in this study, but the observed decreases in, sperm count were neither strictly dose dependent nor consistent between individuals treated with the same dose. Basal leuteinizing hormone (LH) and follicle stimulating hormone (FSH) became undetectable 2 weeks after the start of 250 and 500 mg/wk TC injections and were lost within 5 to 6 weeks of starting 100 mg doses. Pituitary gonadotropin responses to leutinizing hormone releasing hormone (LHRH) disappeared more slowly with FSH responses being lost 1 to 3 weeks after the loss of basal FSH activity. Leuteinizing hormone responses to LHRH appeared to be suppressed last, disappearing 4 to 6 weeks after FSH responses to LHRH. CONCLUSIONS: Exogenous testosterone-mediated inhibitory influences on the hypothalamic-pituitary-testicular axis were reversed following the cessation of drug treatment. PMID- 9394097 TI - SPECT-evaluation of the monoamine uptake site ligand [123I](1R)-2-beta carbomethoxy-3-beta-(4-iodophenyl)-tropane ([123I]beta-CIT) in untreated patients with suspicion of Parkinson disease. AB - BACKGROUND: For a few years, data on SPECT-imaging of dopamine transporters with the cocaine derivate [123I](1R)-2-beta-carbomethoxy-3-beta-(4-iodophenyl)-tropane ([123I] beta-CIT) have been reported mostly in healthy subjects or animals. This study reflects our preliminary results with SPECT-imaging of dopamine transporters using the cocaine analogue 123-beta-CIT in patients with untreated (de novo) parkinsonism. METHODS: In 33 patients with clinical suspicion of Parkinson disease and 5 healthy controls, SPECT-imaging of dopamine transporters was performed 1, 4, and 24 hours after injection of 180 MBq of 123I-beta-CIT, which was generated by iododestannylation. None of the patients or controls had been treated before with neuroleptical drugs or any other pharmaceuticals with known binding to the dopamine transporters. Clinical symptoms were staged by the scales Hoehn-Yahr (HYS), Unified Parkinson Disease Rating Scale (UPDRS), and the self-rating scale of Beck depression inventory (BDI). For evaluation, striatal/cerebellar ratios were calculated to every time point. RESULTS: Significant correlations of 123I-beta-CIT uptake could be stated compared to UPDRS, HYS, and BDI values (Spearman correlation, p < 0.05). The symptoms of rigor and akinesia showed a significant correlation with the beta-CIT uptake, whereas the symptom of tremor failed, which may be caused by the location of tremor symptoms out of the striatum. Comparing the controls, a significant (p < 0.01) decrease of tracer uptake in parkinsonian patients is stated on the images at 24 hours p.i. In our patients, tracer uptake does not depend significantly on duration of disease and age. CONCLUSION: 123I-beta-CIT seems to be a promising tool in imaging of untreated parkinsonian patient. PMID- 9394099 TI - Immunophenotyping as a diagnostic tool to differentiate lichen planus from chronic graft-versus-host disease: diagnostic observations on two patients. AB - BACKGROUND: Lichen planus (LP) and the lichenoid variant of chronic graft-versus host disease (cGVHD) can present with similar clinical and histological findings. The distinction, although difficult, is important both prognostically and therapeutically. The mechanism and effector cell phenotypes have also shown to differ between the 2 entities. While the lichenoid infiltrate of LP is predominantly T lymphocytes helper/inducer cell phenotype, the suppressor/cytotoxic subset appears to play a major role in cGVHD. The aim of this study is to determine whether the immunophenotypic character of the lichenoid infiltrate can aid in distinguishing the 2 entities. METHODS: Biopsies were obtained from 2 patients with lichenoid papules and a history of transplantation. Light microscopy revealed lichenoid inflammation in both cases characterized by a band-like lymphohistiocytic infiltrate at the dermal-epidermal junction. Immunochemistry was performed on fresh tissue using a panel of monoclonal antibodies including anti-CD1a, CD3, CD4, CD8, CD16, CD20, CD28, and CD68. Results were quantitated using computer-assisted image analysis. RESULTS: We found that in both cases the majority of cells stained with pan T cell marker CD3+. One case demonstrated predominantly CD4+ T cells and increased numbers of CD1a positive Langerhans cells, while the lymphokine natural killer cell activity (LAK) markers anti-CD16 and anti-CD28 were largely nonreactive. Conversely, the second case contained predominately CD8+ lymphocytes and very few CD1a positive Langerhans cells with abundant LAK cell anti-CD16 and anti-CD28 reactivity. CONCLUSIONS: Based on these findings, the former was classified as lichen planus and the latter as lichenoid cGVHD. The diagnoses are substantiated with clinical history and follow-up information. We conclude that immunophenotypic characteristics of the infiltrate can be a useful tool in differentiating lichenoid cGVHD from lichen planus. PMID- 9394098 TI - Cardiac high-energy phosphate metabolism in patients with aortic valve disease assessed by 31P-magnetic resonance spectroscopy. AB - BACKGROUND: The purpose of this work was to determine the clinical and hemodynamic correlates of alterations in cardiac high-energy phosphate metabolism in patients with aortic stenosis and with aortic incompetence. METHODS: Fourteen volunteers, 13 patients with aortic stenosis, and 9 patients with aortic incompetence were included. Patients underwent echocardiography and left and right heart catheterization. 31P-MR spectra from the anterior myocardium were obtained with a 1.5 Tesla clinical MR system. RESULTS: Aortic stenosis and aortic incompetence patients had similar New York Heart Association (NYHA) classes (2.77 +/- 0.12 vs 2.44 +/- 0.18), ejection fractions (normal), left ventricular (LV) end-diastolic pressures, and LV wall thickness. In volunteers, phosphocreatine/adenosine triphosphate (ATP) ratios were 2.02 +/- 0.11. For all patients, phosphocreatine/ATP was significantly reduced (1.64 +/- 0.09; *p = 0.011 vs volunteers). Phosphocreatine/ATP decreased to 1.55 +/- 0.12 (*p = 0.008) in aortic stenosis, while in aortic incompetence, phosphocreatine/ATP only showed a trend for a reduction (1.77 +/- 0.12; p = 0.148). For all patients, phosphocreatine/ATP decreased significantly only with NYHA class III (1.51 +/- 0.09; *p = 0.001), but not with NYHA classes I and II (phosphocreatine/ATP 1.86 +/- 0.18). In aortic stenosis, phosphocreatine/ATP ratios decreased (1.13 +/- 0.03; *p = 0.019) only when LV end-diastolic pressures were > 15 mm Hg or when LV diastolic wall stress was > 20 kdyne cm-2 (1.13 +/- 0.03; *p = 0.024). CONCLUSIONS: For a similar clinical degree of heart failure in human myocardium, volume overload hypertrophy does not, but pressure overload does, induce significant impairment of cardiac high-energy phosphate metabolism. In aortic valve disease, alterations of high-energy phosphate metabolism are related to the degree of heart failure. PMID- 9394100 TI - The need for inotropic support in a subgroup of infants with severe life threatening respiratory syncytial viral infection. AB - BACKGROUND: We experienced an unusual complication of life-threatening respiratory syncytial viral disease cardiovascular compromise. Life-threatening respiratory syncytial virus (RSV) infection has predominancy involved with ventilatory support for respiratory distress and/or failure. We performed a retrospective chart review of 20 consecutive infants admitted to the pediatric intensive care unit (PICU) for impending respiratory failure. METHODS: Seventeen required ventilatory support. As part of the infants' initial assessment, blood pressure, distal perfusion [capillary refill time (CRT) > or = 3 sec], decreased peripheral pulses, and peripheral mottling were used to determine cardiovascular compromise. These infants received volume resuscitation either at the referring facility or the PICU until euvolemia was obtained, as determined by central venous pressure (CVP) monitoring (between 3 to 7 cm H20). Nine of the 20 infants did not respond to volume resuscitation alone and required vasopressor support in the form of: Dopamine (7 patients, 5-10 micrograms/kg/min), Dobutamine (2 patients, 5-7 micrograms/kg/min), and one who expired required both Epinephrine (600 ng/kg/min) and Dopamine (10 micrograms/kg/min). The mean ages of these 9 patients were 6.2 +/- 3.4 weeks (range 3-12 weeks), the mean duration of ventilation was 7.2 +/- 4.1 days (range 4-12 days). The mean duration of pharmacologic support was 69.7 +/- 47 hours (range 14-168 hours). The mean ages of RSV+ infants not requiring inotropic support was 19.4 +/- 27.4 weeks (range 1 90 weeks), and mean duration of ventilation was 5.5 +/- 5.9 days (range 2-20 days). RESULTS: The inotrope treated patients were weaned from pharmacologic support prior to extubation, without any hemodynamic deficits. Our experience with this rather high incidence of hemodynamic complications during this RSV epidemic was unexpected. CONCLUSION: These results substantiate the fact that younger patients with RSV disease are at both greater risk for pulmonary complications and cardiovascular deterioration and may thus benefit from pharmacologic support. PMID- 9394101 TI - Interleukin-1 response to arterial antigen, lipopolysaccharide, and oxidized low density lipoprotein in ischemic heart disease. AB - Monocytes responding to oxidized low density lipoprotein (LDL) or other antigens may initiate atherogenesis through production of interleukin-1 (IL-1) and additional cytokines. Interleukin-1 is chemotactic for circulating leukocytes, can stimulate growth of fibroblasts or smooth muscle cells, and causes activation of T- and B-lymphocytes. METHODS: Peripheral blood mononuclear cells (PBMCs) were isolated from 42 patients with angiographically verified ischemic heart disease (IHD) and 35 age-matched control subjects without a history of cardiac disease. Rates of proliferation and production of IL-1 beta were measured after peripheral blood mononuclear cells were cultured for 7 days in the presence of mitogens, arterial antigen, lipopolysaccharide, or native and oxidized forms of LDL. RESULTS: In patients with IHD, proliferation in response to arterial antigen was either diminished or unchanged from control values. Peripheral blood mononuclear cells from IHD and control patients had similar levels of proliferation after treatment with different mitogens. Levels of IL-1 beta, produced after stimulation with arterial antigen or lipopolysaccharide, also did not differ for PBMCs obtained from control and IHD patients. For patients with either a stable angina pattern or no history of cardiac disease, PBMC cultured in the presence of native and oxidized forms of LDL released similar amounts of IL-1 beta. In contrast, PBMCs from 4 patients with unstable angina had increased levels of IL-1 beta after culture in the presence of oxidized LDL (group means +/- standard deviation of 1.63 +/- 1.08 pg/mL for 17 control patients, 0.96 +/- 0.23 pg/mL for 4 cases with stable angina, and 4.02 +/- 5.91 pg/mL, for 19 cases with unstable angina). These values reflect a greater than 5-fold increase in variability for IL-1 beta produced on exposure to oxidized LDL for patients with unstable angina relative to control patients. CONCLUSIONS: Effects of in vitro stimulation with mitogens or lipopolysaccharide are similar for PBMC obtained from normal or IHD patients. The response to arterial antigen is also not increased in cells from patients with IHD. However, PBMCs obtained from a subset of patients with unstable angina produce greater levels of IL-1 beta after treatment with oxidized, but not native, LDL. PMID- 9394102 TI - Gastric myoelectrical activity, gastric emptying and correlations with dyspepsia symptoms in patients with gastroesophageal reflux. AB - BACKGROUND: Delayed gastric emptying is a mechanism that contributes to the pathogenesis of gastroesophageal reflux. Electrogastrogram changes, gastric emptying rates, and Helicobacter pylori status were investigated, and a correlation was sought with dyspepsia symptoms in gastroesophageal reflux disease patients. METHODS: Fifty patients (27 females; mean age 43) with gastroesophageal reflux were studied. Electrogastrographic recordings were obtained 30 minutes before and simultaneously with a 2-hour radionuclide gastric-emptying test using an isotope-labeled solid meal. Symptoms of nausea, abdominal bloating, abdominal pain, and early satiety were graded from 0 to 5. RESULTS: Thirty-six percent of patients had delayed gastric eliminating. Thirty-eight percent (19/50) patients had abnormal electrogastrograms, and 11 of these 19 also had delayed gastric emptying. There was a significant difference in the electrogastrographic parameter of postprandial power change in patients with delayed versus normal gastric emptying (0.20 +/- 0.8 dB vs 3.17 +/- 0.8 dB, p < 0.05). In patients with an abnormal electrogastrogram, the mean symptom score was significantly higher than in patients with a normal electrogastrogram (2.18 +/- 0.26 vs 1.35 +/- 0.16, p < 0.05). Twenty-one percent (7/33) of patients were positive (+) for Helicobacter pylori overall, but this did not seem to affect electrogastrogram and gastric emptying findings. CONCLUSIONS: Fifty-two percent of gastroesophageal reflux disease patients have gastric motor or myoelectrical abnormalities that contribute to the pathogenesis of this entity and also help explain the high prevalence of dyspepsia in the clinical presentation of gastroesophageal reflux disease. PMID- 9394103 TI - Differences in the course of alcohol withdrawal in women and men: a Polish sample. AB - A retrospective study compared the course of alcohol withdrawal, including delirium tremens, in women and men hospitalized in the Nowowiejski Hospital in Warsaw from 1973 to 1987. Medical records pertaining to 1179 patients were analyzed; 13.8% of these patients were women and 86.2% were men. The study showed that women began intensive alcohol drinking later than men (p < 0.0001), but the period between the onset of alcohol abuse and the first occurrence of alcohol withdrawal was shorter in women than in men (p < 0.0001). In the period of heavy drinking before hospitalization, women consumed significantly less alcohol then men (p < 0.0001); moreover, women drank nonbeverage alcohol less frequently than men (p < 0.05). Women were hospitalized substantially longer than men (p < 0.0001), whereas the duration of alcohol withdrawal symptoms at the time of hospitalization was comparable in both groups. Withdrawal seizures were significantly more frequent among men than among women (p < 0.001). Significant differences in the patients' somatic conditions were not noted between the groups, with the exception of anemia and decreased potassium concentration, which were more frequently observed in women (both p < 0.0001), and of increased concentration of ALT and hypoproteinemia, which were more frequent in men (respectively, p < 0.05 and p < 0.01). Co-existing personality disorders, depressive disorders, and anxiety disorders--as well as abuse of benzodiazepines and barbiturates--were more frequently observed in women (p < 0.0001). The period between the first hospitalization due to alcohol withdrawal and the time of death was significantly shorter in men than in women (p < 0.05). The results point to differences in the conditions and the course of alcohol dependence and alcohol withdrawal between women and men. PMID- 9394104 TI - Association analysis of a regulatory variation of the serotonin transporter gene with severe alcohol dependence. AB - The present study tested the hypothesis that the short, low activity variant of a biallelic polymorphism in the 5' regulatory region of the human serotonin transporter (5-HTT) gene confers susceptibility to severe alcohol dependence marked by severe withdrawal symptoms. Applying a phenotype-genotype strategy, our population-based association analysis included 216 German controls and an extreme sample of 103 severely affected alcoholics who were selected from 315 German alcohol-dependent subjects by a history of alcohol withdrawal seizure or delirium. The frequency of the short allele (S) was significantly increased in the severely affected alcoholics, compared with that in the controls (X2 = 3.87, df = 1, nominal p = 0.049). The post-hoc exploration indicated that this allelic association resulted exclusively from a significant excess of the S/S genotype in the severely affected alcoholics (p = 0.035), suggesting a recessively acting effect. Consistently, we found a weak but significant correlation (p = 0.013) between the frequency of the S/S genotype and severity of withdrawal symptoms (WDS): no WDS [18.3%, odds ratio (OR) = 1.16], vegetative WDS only (21.8%, OR = 1.44), and severe WDS with either withdrawal seizure only or delirium only (25.0%, OR = 1.69), and both withdrawal seizure and delirium (30.8%, OR = 2.30). Further studies are required to test whether the tentative genotype-phenotype relationship occurred by chance or reflects a real genotypic association between a recessively modifying effect of the short variant of the functional 5-HTT promoter polymorphism and alcohol withdrawal vulnerability. PMID- 9394105 TI - Gastric ethanol metabolism and gastritis: interactions with other drugs, Helicobacter pylori, and antibiotic therapy (1957-1997)--a review. AB - The stomach provides some protection against the penetration of ethanol into the body by contributing to the metabolism of ethanol. The latter is attenuated by various drugs and, although the magnitude of this effect is still the subject of debate, patients should be warned of the corresponding possible increase in blood alcohol levels. Furthermore, oxidation of ethanol generates acetaldehyde, a toxic metabolite. In addition, chronic alcohol abuse seems to favor colonization by Helicobacter pylori, which produces ammonia that also contributes to the commonly associated chronic gastritis. Because antibiotics were shown over the last 4 decades to effectively eliminate gastric ammonia, they should be considered for the routine treatment of such chronic gastritis in the way they are now being used for ulcer therapy. PMID- 9394106 TI - Effect of treatment with paromomycin on endotoxemia in patients with alcoholic liver disease--a double-blind, placebo-controlled trial. AB - The results of experimental and clinical studies support the hypothesis that gut derived endotoxins might be of relevance for the development and course of alcoholic liver disease. The aim of this study was to test the effect of a nonabsorbable, broad-spectrum antibiotic on endotoxemia in patients with alcoholic liver disease. Fifty patients with alcoholic liver disease (27 with cirrhosis, 23 without cirrhosis) were randomly assigned to receive either paromomycin sulfate (3 x 1 g/day) or placebo in a double-blind fashion for at least 3 weeks, and if possible 4 weeks. Endotoxin concentration, liver function tests, and other laboratory parameters were determined in weekly intervals. Endotoxin concentration was also determined in 15 healthy controls. Groups receiving paromomycin or placebo were similar for clinical and biological items collected initially. Mean initial endotoxin concentrations were significantly elevated in both groups (mean +/- SEM; paromomycin, 16.7 +/- 5.3 pg/ml; placebo, 17.5 +/- 6.9 pg/ml; healthy controls, 2.3 +/- 0.4 pg/ml). Although the mean endotoxin concentration was lower in the verum group after 1 week (paromomycin, 8.0 +/- 1.9 pg/ml; placebo, 14.6 +/- 3.5 pg/ml; p > 0.05), paromomycin treatment had no significant effect on endotoxin concentration or liver function tests during the 4-week period. The beneficial effect of paromomycin treatment on endotoxemia in cirrhotics reported in earlier studies could not be reproduced under the conditions of this trial in patients with alcoholic liver disease. PMID- 9394107 TI - Which relapse criteria best predict the mortality risk of treated alcoholics? AB - We examined which relapse criteria best predict the mortality risk of treated male alcoholics. The subjects were 172 male alcoholics who had previously been hospitalized. Using three criteria which defined relapse as failure to maintain abstinence from alcohol, alcohol abuse, or dependence, the relapse of each subject had been evaluated during a previous 3-year outcome study. Relative mortality risks in the next 3 years classified by the three relapse criteria were compared. The follow-up rate was 93.6% and 31 subjects died. The age-corrected relative mortality risk for subjects failing to maintain abstinence compared with abstainers was 5.32, while the relative mortality risks for the group abusing alcohol and for the group suffering alcohol dependence were 2.23 and 2.56, respectively. These results suggest that relapse defined as failure to maintain abstinence predicts a higher relative mortality risk than do criteria defining in terms of alcohol abuse and alcohol dependence. PMID- 9394108 TI - Predicting problem drinking: a test of an interactive social learning model. AB - This study tested a social learning model and explored the direct and interactive relationships between personality and environment in predicting problem alcohol use. We used longitudinal data from a nonclinical sample of males and females first tested in adolescence and followed into young adulthood. Hierarchial regression analyses were used to test main effects and interaction models. The cross-sectional data supported an interactive social learning model. Both personality and environment variables significantly predicted problem drinking. Two interactions between heavy drinking peer groups and personality variables were significant. Contrary to our hypothesis, the direction of the interaction was negative. In contrast, the longitudinal analyses did not provide strong support for our interactive model. Personality variables were significant predictors longitudinally, but in only one analysis did an environment variable significantly predict problem drinking. Furthermore, none of the interactions was significant predictors over time. Overall, the findings suggest that social learning models based on the interaction of personality and environmental influences may be more appropriate for predicting concurrent, as opposed to future problems, and that future research should include person-environment interactions. In addition, cultural tolerance of heavy drinking may be an important determinant of the role of psychological vulnerability in the development of problem drinking. PMID- 9394109 TI - Comparison of screening instruments for alcohol problems between black and white emergency room patients from two regions of the country. AB - A number of brief screening instruments to identify alcohol dependence exist, but the validity of these instruments across ethnic groups or regions of the country is not well established. The sensitivity and specificity of a number of standard screening instruments (CAGE, brief MAST, AUDIT, TWEAK, and RAPS), as well as other measures (History of Trauma Scale, breathalyzer reading, self-reported drinking before the event, and consuming five or more drinks at a sitting at least monthly) are compared against ICD-10 and DSM-IV criteria for alcohol dependence between probability samples of Black and White emergency room patients in Santa Clara County, CA (n = 716) and in Jackson, MS (n = 1330). Variability in the sensitivity of screening instruments among current drinkers was found to be greater between samples for both Blacks and Whites, than for Blacks compared with Whites within the same sample. The AUDIT, TWEAK, and RAPS seemed to perform well by gender and injury status for both Blacks and Whites in the two samples, and no significant differences were found in the performance of these instruments across sample sites. To evaluate the influence of regional differences in alcohol dependence on differences found in the performance of screening instruments, using logistic regression with the simultaneous entry of demographic variables (age, gender, ethnicity, injury status, and site) and drinking variables (breathalyzer reading, self-reported drinking before the event, and drinking five or more drinks at a sitting at least monthly) to predict alcohol dependence in a merged sample of these patients (Jackson vs. Santa Clara) site was not found to be significant. Data suggest that, whereas region of the country may not be important in predicting alcohol dependence in emergency room populations, regional differences in the performance of screening instruments for alcohol dependence may exist, even when ethnicity is taken into account. Given distinct regional differences in drinking patterns and problems in the U.S., further research on commonly used screening instruments is needed to determine those screeners most efficient for identifying problem drinking. PMID- 9394110 TI - Neurophysiological correlates of response production and inhibition in alcoholics. AB - Scalp recordings of the P300 component of the event-related potential were made from a group of medication-free, chronic male alcoholics and a control group, participating in a visual Go/No Go reaction time paradigm. Subjects were presented with large and small forms of the letters T and V. The large forms (Go stimuli) required a button press with either the left or right hand, whereas the small forms (NO Go stimuli) required response inhibition. Recordings were made from 31 electrodes that, for statistical analyses, were grouped into five regions: frontal, central, parietal, occipital, and temporal. The results indicated that, in each of the five regions, both Go and NO Go response amplitudes were larger in the controls than in the alcoholics. No group differences in latency were observed in any region. Surface energy (Wang et al., Brain Topogr. 6:193-202, 1994) magnitudes paralleled P300 amplitudes and in the controls, compared with the alcoholics, were larger during both Go and No Go trials. Our findings suggest that abstinent, chronic alcoholics differ electro physiologically from control individuals. These differences are manifested as widespread reductions in P300 amplitudes during the performance of a simple information processing paradigm. The reduced amplitudes may reflect a deficiency in an inhibitory mechanism proposed to underlie P300 generation. PMID- 9394111 TI - Voices of the afflicted. AB - Over the past 10 years, I have been privileged to conduct educational forums for audiences containing many recovering alcoholics or otherwise chemically dependent persons. In these forums about the addictive diseases and their treatment and research possibilities, significant interaction with the audience members occurs. During these interactions, certain anecdotal phenomena seem to predominate. The repetitive nature of these reports suggests the need for systematic investigation. As with editorial comments in major medical journals, observed phenomena and unanswered questions from those afflicted can be valuable in the generation of testable hypotheses. Perhaps the ideas presented herein will be useful in the development of future research on alcohol abuse and alcohol dependence. PMID- 9394112 TI - Effects of ethanol on rat Sertoli cell function: studies in vitro and in vivo. AB - Chronic alcohol administration to male animals is associated with testicular atrophy and gonadal failure. The Sertoli cell seems to be the first testicular cell injured as a result of alcohol exposure. To investigate the adverse effects of ethanol on testicular and particularly Sertoli cell function, the consequences of in vivo and in vitro ethanol exposure on rat Sertoli cell mRNA and protein levels of transferrin and ornithine decarboxylase were investigated. In vivo, ethanol exposure enhanced the levels of both hepatic and testicular (Sertoli cell) transferrin protein and mRNA. Ethanol exposure also enhanced testicular, but not hepatic, levels of ornithine decarboxylase protein and mRNA. These in vivo findings were confirmed when isolated Sertoli cells were studied in vitro. Specifically, ethanol exposure increased Sertoli cell transferrin protein and mRNA levels. Ethanol exposure increased Sertoli cell ornithine decarboxylase mRNA and protein when cultured in serum-free media, but not when cultured in the presence of serum. These studies demonstrate that ethanol exposure of rat Sertoli cells is associated with alterations in the levels of mRNA and protein that are known to be important in the process of spermatogenesis. These findings add to the body of evidence that suggests that, within the testes, the Sertoli cell may be an important target for ethanol-induced gonadal injury. PMID- 9394113 TI - Fetal alcohol exposure and temporal vulnerability regional differences in alcohol induced microencephaly as a function of the timing of binge-like alcohol exposure during rat brain development. AB - In humans, microcephaly (small head for body size) is a common feature of fetal alcohol syndrome. An analogous measure, termed microencephaly (small brain for body size), can be used for evaluating the detrimental effects of the differential timing of alcohol exposure on brain development in animal model systems. Timed-pregnant rats were exposed to binge-like alcohol during either the first 10 days (first trimester equivalent) or second 10 days of gestation (second trimester equivalent), or the combination of first and second trimesters equivalent for prenatal treatments. Offspring from some of the animals exposed to alcohol during the combined first and second trimesters equivalent were raised artificially from postnatal day (P) 4 through P9 (part of the third trimester equivalent), and also received binge-like alcohol during this period, producing animals that were exposed to alcohol during all three trimesters equivalent. Offspring from untreated dams were also raised artificially and received alcohol only from P4 to P9, thus creating animals that were exposed to alcohol only during part of the third trimester equivalent. All pups were perfused on P10. Appropriate controls (nutritional and normally reared) were used for every alcohol treatment combination. Peak blood alcohol concentrations were not different among the treatment groups for a given sampling time. Significant somatic growth deficits occurred in offspring exposed to alcohol for the equivalent of all three trimesters, compared with offspring exposed to alcohol during other periods. Brain growth in offspring also was significantly affected by the timing of alcohol exposure. The whole brain, forebrain, and cerebellum to body weight ratios of pups exposed to alcohol during the third trimester had more significant brain growth deficits than pups in groups exposed to alcohol during other times of brain development. Although alcohol exposure during the third trimester had a significant detrimental impact on overall brain growth, significant differences in temporal vulnerability were also found for the brainstem to body weight ratios. Offspring of dams exposed to alcohol during the first trimester had the same magnitude of deficit as those exposed to alcohol during the third trimester, and those two groups were significantly deficient compared with the groups exposed to alcohol at other times, suggesting some differential vulnerability of this region to alcohol-induced injury at different times of development. This study is the first thorough examination of microencephaly and gross regional vulnerability of the developing brain as related to temporal factors of alcohol exposure in an animal model system, and the results support and expand on the findings of the available clinical literature. Furthermore, our results substantiate claims that the cessation of alcohol before the third trimester can lessen the severity of some alcohol related birth deficits. PMID- 9394114 TI - Ethanol feeding causes inactivation of both state 1 and state 2 rat hepatic asialoglycoprotein receptors. AB - Previous studies have shown that ethanol feeding in rats causes inactivation and redistribution of approximately 50% of the total asialoglycoprotein receptors (ASGPRs) in hepatocytes (Tworek et al., J. Biol. Chem. 271:2531, 1996), and that two equal populations of hepatic ASGPRs mediate ligand uptake and processing via two functionally different pathways (Weigel in Glycoconjugates: Composition, Structure and Function, Marcel Dekker, 1992, p. 421). The purpose of this study was to determine if ethanol feeding causes preferential inactivation of only one of these two ASGPR populations, which have been designated state 1 and state 2 ASGPRs. The state 2, but not state 1, ASGPRs are inactivated in isolated hepatocytes by a variety of drugs and inhibitors. State 2 ASGPRs can also be inactivated in permeable cells by ATP treatment and then reactivated by treatment with fatty acyl coenzyme As. In the present study, permeable cell assays for state 2 ASGPR inactivation and reactivation were used to assess whether hepatocytes from ethanol-fed rats contain inactive state 2 ASGPRs. The results show that preferential inactivation of one ASGPR population does not occur after ethanol feeding. That inactive ASGPRs could not be reactivated by treatment with palmitoyl-coenzyme A to a greater extent in ethanol-fed versus control cells indicates there is not a larger pool of inactivated state 2 ASGPRs in treated cells. We conclude that ethanol feeding causes equal inactivation of both state 1 and state 2 ASGPRs. Ethanol feeding may represent the first treatment found to inactivate state 1 ASGPRs. PMID- 9394115 TI - Combination of naltrexone and fluoxetine on rats' propensity to take alcoholic beverage. AB - Naltrexone (NTX) and fluoxetine (FLU) are useful for treating alcoholism and depression, respectively. Furthermore, these afflictions covary. Given the possibility that people might be prescribed NTX and FLU concurrently, we assessed the effects of these two agents on rats' propensity to drink an alcoholic beverage. Rats were given 65 days of access to a sweetened alcoholic beverage and water for 2 hr daily. At first, they took little ethanol, but after 20 days, they took on average 2.0 to 2.5 g/kg of ethanol, daily during the 2-hr session. They also took sufficient water to maintain their health. After 30 days, they were divided into four groups to receive, 30 min before the drinking session, 1 of 4 different kinds of injections. For the next 20 days, one group received placebo daily. Another group received 5 mg/kg of NTX daily and another 5 mg/kg of FLU daily. The fourth group received both 5 mg/kg of NTX and 5 mg/kg of FLU daily. After 20 days, the doses of NTX and FLU were doubled across an additional 10 days. Both NTX and FLU reduced rats' intake of alcoholic beverage. The combinations of NTX and FLU, however, were no more effective in reducing rats' intake of alcoholic beverage than either alone. Also, the small dose of NTX seemed to lose its effectiveness with repeated administrations. A second experiment confirmed the conclusion that small doses of NTX lose their effectiveness in suppressing intake of alcoholic beverage across repeated administrations. In summary, data provide no support for the idea that FLU and NTX would act synergistically to reduce propensity to take alcoholic beverages. PMID- 9394116 TI - The effect of cold stress on lymphocyte proliferation in fetal ethanol-exposed rats. AB - Prenatal ethanol exposure and stress have each been shown to have significant effects on the immune system. This study examined the possible interactive effects of prenatal ethanol exposure and exposure to stress later in life on the immune system. Differential vulnerability to these challenges in female and male offspring was assessed. At 5 to 6 months of age, female and male offspring from prenatal ethanol-exposed (E), pair-red (PF), and ad libitum-fed control (C) conditions were exposed to 0, 1 or 3 days of cold (4 degrees C). At the end of the cold period, the proliferative response of splenic lymphocytes to the mitogens concanavalin A (Con A) and pokeweed mitogen (PWM) was assessed. The data demonstrate a significant interactive effect between prenatal ethanol exposure and cold stress in female offspring. After 1 day of cold stress, E females had significantly increased PWM-induced lymphocyte proliferation compared with PF and C females, and significantly increased Con A-induced lymphocyte proliferation compared with PF females. There were no differences in PWM or Con A-induced lymphocyte proliferation among E, PF, and C females after 0 or 3 days of cold stress, nor among E, PF, and C males on any test day. Regardless of prenatal treatment, females exposed to 1 or 3 days of cold had significantly greater basal plasma corticosterone levels than females not exposed to cold. In contrast, only E males exposed to 1 or 3 days of cold had significantly increased basal plasma corticosterone levels, compared with E males not exposed to cold; PF and C males showed no significant change in basal corticosterone after cold stress. These data demonstrate that, in response to the challenge of cold stress, changes in lymphocyte proliferation to PWM and Con A may occur selectively in E females. Moreover, the interactive effects of prenatal ethanol and cold stress may result in enhanced rather than suppressed immune responsiveness. PMID- 9394117 TI - Selective inhibition of alcohol intake in diverse alcohol-preferring rat strains by the 5-HT2A antagonists amperozide and FG 5974. AB - The present studies sought to elucidate the role of 5-HT2A receptor antagonists in suppressing alcohol intake by comparing the effects of amperozide and FG 5974 on alcohol, food, and water intake in strains of alcohol-preferring rats: P, Alko Alcohol (AA), and Fawn-Hooded (FH). Both amperozide and FG 5974 have 5-HT2A receptor antagonist properties, but FG 5974 also shows presynaptic 5-HT1A receptor agonist activity. After establishment of stable baselines for intake measures in a two-bottle continuous access paradigm, rats (n = 10) were injected with 1 of 5 doses (0, 2.5, 5.0, and 10.0 mg/kg, sc) of amperozide or FG 5974 at weekly intervals. Amperozide dose-dependently reduced alcohol intake, total fluid intake, and alcohol preference in all three strains under continuous access conditions, whereas FG 5974 was less effective. Food intake was also suppressed by amperozide at higher doses, whereas it was increased by FG 5974. Amperozide also dose-dependently reduced alcohol intake when it was available for only 1 hr/day, but FG 5974 tended to increase it. After oral administration, amperozide was also more effective than FG 5974 in reducing alcohol intake. Despite these differences in efficacy in suppressing alcohol intake, both compounds produced taste aversion to a novel saccharin solution. These complex findings suggest that biochemical properties other than 5-HT2A receptor antagonism (e.g., 5-HT1A receptor agonism) may be involved in the effects of amperozide and FG 5974 on alcohol intake and other consummatory behaviors. PMID- 9394118 TI - Content of dynorphins and kappa-opioid receptors in distinct brain regions of C57BL/6 and DBA/2 mice. AB - Differences in the activity of various components of the endogenous opioid system under basal conditions and after ethanol exposure have been reported between strains and lines of animals showing either high or low ethanol consumption. The objective of the present studies was to investigate the presence of differences in (a) the density of kappa-opioid binding sites, (b) the content of prodynorphin mRNA, and (c) the content of dynorphin peptides in distinct brain regions between the C57BL/6 (ethanol-preferring) and the DBA/2 (ethanol-avoiding) mice. Results indicated that the C57BL/6 mice have a higher content of kappa-opioid binding sites and dynorphin A 1-13 in the amygdala, and dynorphin A 1-8 in the ventral tegmental area, whereas the DBA/2 mice presented a significantly higher content of kappa-opioid binding sites, prodynorphin mRNA, as well as dynorphin A 1-13 and dynorphin A 1-8 peptides in the nucleus accumbens and septum. In addition, the DBA/2 mice presented a higher content of kappa-opioid receptors in the periaqueductal gray and dynorphin A 1-13 and dynorphin A 1-8 in the caudate putamen. Because enhanced stimulation of the kappa-opioid receptors at the level of the nucleus accumbens has been associated with decreased dopamine release and aversive states, the higher content of kappa-opioid receptors, pro-dynorphin mRNA, and dynorphin peptides (the endogenous ligand of k-binding sites) in regions of the limbic system of the DBA/2 mice may play an important role in determining their low alcohol consumption. PMID- 9394119 TI - Effects of chronic ethanol on the mobilization of arachidonate and docosahexaenoate stimulated by the type 2A serotonin receptor agonist (+/-)-2,5 dimethoxy-4-iodoamphetamine hydrochloride in C6 glioma cells. AB - We studied the effects of chronic ethanol exposure on the mobilization of polyunsaturated fatty acids stimulated by activation of the type 2A serotonin receptor in C6 glioma cells. In our in vitro model, we prelabeled cells with [3H]arachidonate and [14C]docosahexaenonate and subsequently stimulated with the type 2A serotonin receptor agonist (+/-)-2,5-dimethoxy-4-iodoamphetamine hydrochloride. In as early as 10 days of exposure to 20 or 50 mM ethanol, the (+/ )-2,5-dimethoxy-4-iodoamphetamine hydrochloride-simulated mobilization of [3H]arachidonic acid [[3H]AA) and [14C]docosahexaenoic acid ([14C]DHA) was significantly inhibited, and this inhibition was accompanied by decreased mobilization of intracellular [Ca2+]i. Exposure to ethanol did not alter significantly the release of [3H]AA and [14C]DHA stimulated by the calcium ionophore A23187 nor the incorporation of [3H]AA and [14C]DHA into cellular lipids. Decreased mobilization of polyunsaturated fatty acids and calcium in astroglia may contribute to neurotoxicity caused by chronic ethanol exposure. PMID- 9394120 TI - Ethanol increases surface-localized fibrinolytic activity in cultured endothelial cells. AB - Epidemiological studies demonstrated a positive association between moderate alcohol consumption and reduced cardiovascular mortality that may be mediated, in part, through increased fibrinolysis. These studies were conducted to determine whether low concentrations of alcohol (0.025 to 0.1%, v/v) directly affected the surface-localized versus secreted/solution phase fibrinolytic activity in live cultured endothelial cell (EC) types. Confluent live cultured ECs [human umbilical vein ECs (HUVECs), human saphenous vein ECs (HSVECs), and porcine aortic ECs (PAECs)] were preincubated (0 to 20 min, 4 degrees C) in the absence or presence of varying concentrations of alcohol (0 to 0.1%, v/v), in the presence of saturating levels of 125I-labeled Glu-plasminogem (2 microM) and 125I Plasmin M(r) 20-kDA light-chain formation quantitated by phosphorimaging autoradiography analysis. Endogenous plasminogen activator (PA)-mediated fibrinolytic activity was time- and dose-dependent; reached a maximum approximately 5- to 10-fold increase at 0.05% alcohol in HUVECs, HSVECs, and PAECs; was completely inhibited by anti-t-PA IgG in HUVECs; and partially inhibited by both anti-t-PA (approximately 40%) and anti-u-PA IgG (approximately 60%) in HSVECs. Complete inhibition of alcohol-induced (0.05%) fibrinolytic activity in cultured HUVECs by 2 mM tranexamic acid (an antagonist of plasminogen binding) indicated that the increased fibrinolytic activity was receptor-bound and localized to the EC surface, rather than present in or secreted into the medium (solution phase). Finally, the alcohol-induced increased fibrinolytic activity in cultured HUVECs returned to essentially normal control levels in approximately 1 hr. These studies have demonstrated a direct effect of low alcohol on EC fibrinolytic activity that may contribute, in part, to the decreased risk for thrombosis, coronary artery disease, and myocardial infarction associated with moderate alcohol consumption. PMID- 9394121 TI - The density of monoamine oxidase B sites is not altered in the postmortem brain of alcoholics. AB - The status of the enzyme monoamine oxidase B (MAO-B) was directly evaluated in the postmortem brain from 20 alcoholics and 23 matched controls. The density of MAO-B sites was quantified by the specific binding of the selective inhibitor [3H]Ro 19-6327 (lazabemide) (8 nM) to cortical membranes. A positive correlation between age at death and MAO-B density was observed in the total sample (r = 0.37, p = 0.015). The density of MAO-B in alcoholics (Bmax = 1,263 +/- 131 fmol/mg of protein) was not different form that in control (Bmax = 1,131 +/- 96 fmol/mg of protein). Ethanol in vitro inhibited [3H]Ro 19-6327 binding, with similar potency in membranes form alcoholics (Ki = 280 +/- 13 mM) and controls (Ki = 338 +/- 84 mM). The present results in brain tissue contrast with previous reports of decreased MAO-B enzymatic activity in platelets of alcoholics, but strongly agree with recent genetic studies on MAO-B status in alcoholism. PMID- 9394122 TI - Regional restriction of alcohol/retinol dehydrogenases along the mouse gastrointestinal epithelium. AB - The gastrointestinal tract is a major site of alcohol dehydrogenase (ADH) activity in humans and rodents. Because class I ADH (ADH-I) and class IV ADH (ADH IV), but not class III ADH (ADH-III), function as retinol dehydrogenases in vitro and may thus participate in retinoid signaling needed for epithelial differentiation, the aim of this study was to determine the localization of these enzymes along the gastrointestinal tract. Specific antibodies were used to examine the tissue distribution of all three known classes of mouse ADH by Western blotting, and cellular localization was determined by immunohistochemistry. ADH-I was detected primarily in the intestine, liver, kidney, adrenal, and uterus, with detection of ADH-III in all tissues examined, and detection of ADH-IV primarily in the esophagus, stomach, adrenal, skin, ovary, and epididymis. Along the gastrointestinal tract, ADH-III was not specifically localized, whereas ADH-I was localized exclusively in the villus epithelium of the small intestine and absorptive epithelium of the large intestine, with ADH-IV being localized exclusively in the basal and suprabasal epithelial cells of the esophagus and gastric pit surface epithelium of the stomach. The ADH localization patterns observed are consistent with ADH-I and ADH IV, but not ADH-III, functioning physiologically in retinol metabolism needed for epithelial differentiation. Our results further suggest that the functions of ADH I and ADH-IV are regionally restricted to the lower and upper components, respectively, of the gastrointestinal epithelium, a finding that may relate to the different efficiencies of these two enzymes for retinol oxidation, as well as to the different susceptibilities of the upper and lower digestive tracts for ethanol-induced cancers. PMID- 9394123 TI - Differences in free-choice ethanol acceptance between taste aversion-prone and taste aversion-resistant rats. AB - Taste aversion (TA)-prone (TAP) and TA-resistant (TAR) rats were tested for naive, nonforced acceptance of ethanol. Ethanol acceptance had played no role in line development. Rather, the lines had been developed via bidirectional, nonsibling matings based on susceptibility to develop cyclophosphamide-induced conditioned TAs to a 0.1% saccharin solution (at cyclophosphamide doses of 12.5 mg/kg for males and 15.0 mg/kg for females, i.p.). Rats from the 23rd selectively bred generations, with no prior exposure to ethanol, were given 24-hr access to a two-bottle choice between plain tap water and a solution of ethanol in water. Rats were initially given access to 1% ethanol in water, and the ethanol concentration was increased by 1% every 3 days to a maximum of 10%. Ethanol consumption (g ethanol consumed/kg body weight) and preference scores (volume ethanol solution consumed/total fluid intake) were determined by daily bottle weighings. At 1% ethanol concentration, there were no differences between the rat lines in terms of ethanol consumption or preference. At concentrations of 2 to 10%, TAP rats consumed less ethanol and showed a decreased preference for the ethanol solutions than TAR rats. Maximum ethanol consumption was reached at the 6% concentration for both lines. The mean (+/- SE) values of consumption at 6% ethanol were 1.8 (+/- 0.8) and 5.6 (+/- 0.5) g of ethanol/kg body weight for TAP and TAR rats, respectively. Mean (+/- SE) preference scores at 6% ethanol were 26 (+/- 12) and 76 (+/- 6) for TAP and TAR rats, respectively. These findings indicate that differences in TA conditionability may be associated with the propensity of rats to be high or low consumers of ethanol. Based on these data, it is hypothesized that high susceptibility for TA conditionability may deter many individuals from consuming the high levels of ethanol that usually precede alcohol tolerance and dependence. PMID- 9394124 TI - Genetics ethanol and the Fos response: a comparison of the C57BL/6J and DBA/2J inbred mouse strains. AB - The effect of ethanol (0.25 to 4 g/kg) on the number of Fos-like immunoreactive (Fos-li) neurons was studied in the C57BL/6J (B6) and DBA/2J (D2) inbred mouse strains. The brain regions emphasized in the analysis were from the basal ganglia and some associated limbic nuclei. The question addressed was whether or not the D2 and B6 strains differed in these regions in a way that could explain the marked psychomotor stimulation of the D2, but not the B6, strain over the dose range of 1 to 2 g/kg of ethanol. Over the dose range of 0.25 to 2 g/kg, ethanol caused a modest increase in the number of Fos-li neurons within the caudate putamen (dorsolateral and dorsomedial) and the nucleus accumbens (core and shell), but there were no marked strain effects. There was no significant effect in either strain of ethanol treatment (0.25 to 2 g/kg) in the globus pallidus, ventral pallidum, and subthalamic nucleus. However, at 4 g/kg, there was a dramatic (> 100%) increase of Fos-li neurons in the D2 but not B6 strain. A similar effect was noted in the entopeduncular nucleus, the substantia nigra zona reticulata (and compacta), but not the ventral tegmental area. A marked and substantial (> 200%) Fos response was seen in the central amygdaloid nucleus (CeA) of the D2 strain over the entire dose range; in contrast, a substantial Fos response in the B6 strain was seen only at the 4 g/kg dose. The paraventricular thalamic nucleus, in general, paralleled data in the CeA; but, the Fos response was more modest, and the results for the D2 strain were significant only at the 2 g/kg dose. Overall, data suggest that ethanol at low to moderate doses induces significant, strain-dependent Fos responses in some limbic structures, but not in the basal ganglia. The possibility is considered that activation of some neurons in the CeA are permissive for expression of the ethanol-induced increase in motor activity. PMID- 9394125 TI - NMDA receptor binding in adult rat brain after several chronic ethanol treatment protocols. AB - The amino acid L-glutamate is a major excitatory neurotransmitter that is involved in many CNS functions, including learning, memory, long-term potentiation, and synaptic plasticity. Acute exposures to ethanol (50 to 200 mM) have been shown to inhibit NMDA receptor responses, whereas chronic exposure to ethanol leads to adaptive supersensitivity thought to be involved in ethanol dependence and tolerance. To investigate the effects of chronic ethanol exposure on glutamate receptor density, we examined the binding of both NMDA and non-NMDA ligands in rat brain after several chronic ethanol treatment protocols using a number of different rat strains. No increases in the binding of [3H]MK-801, [3H]CGP 39653, or the polyamine specific competitive antagonist, [3H]ifenprodil, were seen after two well-used chronic ethanol treatments. These included the 2 week liquid diet developed by Frye et al. (J. Pharmacol. Exp. Ther. 216:306-314, 1981) and the 4-day binge treatment developed by Majchrowicz (Psychopharmacologia 43:245-254, 1975). However, small increases in the binding of both the NMDA noncompetitive antagonist [3H]MK-801, as well as the competitive NMDA antagonist [3H]CGP 39653, were seen in select frontal brain regions after 3 weeks of the Walker-Freund chronic ethanol liquid diet. When this chronic liquid diet treatment was extended to a period of 6 weeks, these increases in receptor binding were diminished to nonsignificant levels. The binding of the non-NMDA ligands [3H]AMPA and [3H]kainate were not significantly affected by either length of Walker-Freund liquid diet exposure. When rats were treated chronically with ethanol for 30 days using the paradigm developed by Tsukamoto et al. (Hepatology 5:224-232, 1985), small, but significant, increases in the binding of [3H]MK-801 were seen in the CA1 and dentate gyrus regions of the hippocampus. These studies indicate that robust increases in NMDA receptor binding do not occur with several chronic ethanol treatment protocols, and suggests that NMDA receptor supersensitivity during the development of tolerance and dependence to ethanol may not simply be due to changes in the density of NMDA receptors, but may involve other mechanisms. PMID- 9394126 TI - Chronic ethanol exposure alters leukocyte subsets in repopulating spleens, but does not alter negative selection in thymuses of sublethally irradiated mice. AB - Results from previous in vitro experiments in this laboratory suggested that ethanol may affect selection processes in the thymus. To determine whether ethanol allows escape of potentially autoreactive T-cell clones from negative selection, we fed ethanol to sublethally irradiated, young, adult C57BR mice during the time of thymic and splenic repopulation as a new model of human third trimester fetal alcohol exposure. The mice received a whole-body, sublethal dose (6 Gy) of gamma irradiation at 5 to 6 weeks of age. Feeding of a liquid diet providing 25% of calories as ethanol (EDC) or an isocaloric control liquid diet was begun 3 days after irradiation and was continued for 5 weeks. Each EDC mouse had 2 weight- and age-matched controls, 1 pair-fed (PF), and 1 fed ad libitum (AD LIB). Average blood alcohol concentrations (90 to 440 mg/100 ml) were higher than those reported previously for neonatal mice exposed to ethanol through lactation. At 5 weeks after irradiation, the EDC mice had lower total thymocyte numbers (p < 0.05) and a higher proportion of CD4-CD8-thymocytes than either the PF or AD LIB mice (p < 0.05), which is consistent with findings using in utero models of ethanol exposure. Ethanol exposure also altered the proportion of leukocyte subsets in repopulating spleens. B cells were the most sensitive to the detrimental effects of ethanol and, as a percentage of total nucleated cells in the spleen, B cells were decreased in the EDC group, compared with both the PF and AD LIB groups (p < 0.05). C57BR mice normally delete by negative selection thymocytes bearing v beta 17 T-cell receptors. There was no discernible effect of ethanol exposure during thymic and splenic repopulation on the expression of V beta 17a on thymocytes and splenic T lymphocytes, indicating that ethanol does not affect negative selection. PMID- 9394127 TI - Chronic alcohol ingestion enhances tumor necrosis factor-alpha expression and salivary gland apoptosis. AB - We investigated the extent of induction in sublingual salivary gland cells apoptosis and tumor necrosis factor-alpha (TNF-alpha) expression with chronic ethanol ingestion. The experiments were conducted on rats pair-fed for 8 weeks with alcohol-containing and control liquid diet. The animals were killed, their sublingual glands dissected, and the glandular tissue used for quantization of TNF-alpha expression and the assays of acinar cells apoptosis employing sandwich enzyme immunoassay for histone-associated DNA fragments. The mean value for TNF alpha in sublingual gland of the control group was 22.3 pg/mg of protein and showed a 1.6-fold increase in the chronic ethanol diet group to 36.5 pg/mg of protein. In comparison with the controls, the sublingual gland of the chronic ethanol diet group also exhibited a 3.4-fold enhancement in acinar cell apoptosis. These findings suggest that chronic ethanol ingestion causes the enhancement in TNF-alpha expression and leads to the induction in salivary gland acinar cells apoptosis. Thus, the diminished secretion of saliva in alcoholics may be a direct result of increased salivary gland apoptosis. PMID- 9394128 TI - Effects of prenatal ethanol exposure on phospholipase C-beta 1 and phospholipase A2 in hippocampus and medial frontal cortex of adult rat offspring. AB - Previous studies in our laboratory using a rat model of fetal alcohol exposure (FAE) suggest that FAE-induced behavioral deficits are, in part, linked to neurochemical and electrophysiological deficits in long-term potentiation (LTP) in the entorhinal cortical perforant path projection to the hippocampal formation. Several findings suggest that signal-activated phospholipase C (PLC) and phospholipase A2 (PLA2) are critical to the induction and maintenance of LTP. Thus, alterations in phospholipid metabolism may play a significant role in the LTP deficits observed in FAE offspring. To test this hypothesis, we measured PLC beta 1 and PLA2 activities in the hippocampus and medial frontal cortex of adult rats prenatally exposed to ethanol. PLC-beta 1 activities were significantly decreased by 20 to 30% in both the hippocampus and medial frontal cortex of FAE rats, compared with ad libitum and pair-fed controls. Total Ca(2+)-dependent PLA2 activity was 25% lower in the medial frontal cortex of FAE rats, but did not significantly differ from controls in the hippocampal formation. Approximately 30% of the measured activity in both the medial frontal cortex and hippocampal formation of ad libitum and pair-fed animals was associated with an 85 kDa cytosolic PLA2 form. Cytosolic PLA2 activities were significantly reduced in both the medial frontal cortex and hippocampal formation of FAE rats, compared with controls. These changes in Ca(2+)-dependent PLA 2 and PLC-beta 1 activities, coupled with reports of FAE-induced deficits in protein kinase C activity, indicate that prenatal exposure to moderate quantities of ethanol causes profound and long-lasting deficits in the cellular signaling mechanisms associated with activity-dependent synaptic plasticity and memory formation. PMID- 9394129 TI - Fetal alcohol exposure augments the blunting of tumor necrosis factor production in vitro resulting from in vivo priming with lipopolysaccharide in young adult male but not female rats. AB - We previously reported altered responses of thymocytes and splenocytes to mitogen stimulation in fetal alcohol-exposed (FAE) male Sprague-Dawley rats. We also reported enhanced neuroendocrine responses to stressful stimuli in these animals. The experiments we describe herein aimed at testing whether young adult FAE rats manifest a notable dysregulation in the neuroendocrine-immune response to pathogen administration. We tested the effect of in vivo priming of the animal with a low dose of endotoxin [lipopolysaccharide (LPS), 5 micrograms/kg], considered to be suboptimal from the perspective of mounting detectable levels of circulating monokines several hours after administration, upon the production of immunoreactive tumor necrosis factor (TNF-alpha) in response to a further in vitro challenge of peripheral blood mononuclear cells with 2.5 micrograms/ml of LPS 90 min after priming. We show that the response to the LPS pathogen in vitro after priming is significantly blunted (p < 0.01) in male rats exposed prenatally to alcohol, compared with control male animals. FAE female rats and FAE ovariectomized female rats do not show significant differences in the priming response, compared with control animals. We also show that there is no correspondence between plasma corticosterone levels and TNF-alpha production after priming in any of the groups tested. PMID- 9394130 TI - Alcohol-induced expression of the CD14 endotoxin receptor protein in rat Kupffer cells. AB - Gut-derived endotoxins (lipopolysaccharide, LPS) are believed to contribute to alcohol-induced liver disease (ALD) by stimulating Kupffer cells, the resident liver macrophages, to release proinflammatory cytokines. This activation is largely mediated by CD14, a high-affinity membrane-anchored receptor for LPS. We observed, by chemiluminescence-enhanced detection, an increase in immunoreactive CD14 protein in Kupffer cells isolated from rats treated with ethanol for 2 weeks. Immunocytofluorescence experiments confirmed that this increase was confined to the membranes of Kupffer cells from the alcohol-treated rats. The increase was regulated pretranslationally: a 3-fold elevation (p < 0.01) in the hepatic level of CD14 mRNA was observed. The marked increase in CD14 expression suggests a new mechanism by which alcohol increases the LPS-mediated cytokine signaling by the liver macrophages, thus promoting the interaction between alcohol and endotoxins in the development of liver damage. PMID- 9394131 TI - "Big" versus "little" science: comparative analysis of program projects and individual research grants. AB - Controversy about the amount and nature of funding for mental retardation research has persisted since the creation of NICHD. An issue that has aroused considerable debate, within the mental retardation research community as well as beyond, is distribution of funds between large group research grants, such as the program project (PO1) and the individual grant (RO1). Currently within the Mental Retardation and Developmental Disabilities Branch, more money is allocated to the PO1 mechanism than the RO1. We compared the two types of grants, focusing on success rates, productivity, costs, impact, publication practices, and outcome and conducted a comparative analysis of biomedical and behavioral research. Other related issues were considered, including review processes and cost effectiveness. PMID- 9394132 TI - Maladaptive behavior in children with Prader-Willi syndrome, Down syndrome, and nonspecific mental retardation. AB - Although some genetic, mental retardation syndromes have well-described behavioral features, comparative studies have not yet assessed the relative uniqueness of these so-called phenotypes. Maladaptive behavior of 43 children with Prader-Willi syndrome was compared to age- and gender-matched children with Down syndrome and with nonspecific mental retardation. The Prader-Willi group showed more frequent and severe internalizing, externalizing, and total problem behaviors on the Child Behavior Checklist. Some problems were elevated in all groups, and 12 behaviors were significantly elevated in Prader-Willi subjects relative to both comparison groups. Seven behaviors predicted membership into the Prader-Willi group with 91% accuracy. Implications were discussed for research on behavioral phenotypes in general and for dual diagnosis in particular. PMID- 9394133 TI - Intensive behavioral treatment for preschoolers with severe mental retardation and pervasive developmental disorder. AB - From archival records, we assessed outcomes achieved by preschoolers with both severe mental retardation and autistic features: (a) an experimental group (n = 11), which received intensive behavioral treatment, and (b) a comparison group (n = 10), which received minimal treatment. At intake (mean CA = 3.08 years), the groups did not differ significantly on any variable. At follow-up children in the experimental group obtained a higher mean IQ and evinced more expressive speech than did those in the comparison group. Behavior problems diminished in both groups. Results indicate that intensively treated children achieved clinically meaningful gains relative to the comparison group but remained quite delayed. PMID- 9394135 TI - Fathers and mothers of school-age children with developmental disabilities: parental stress, family functioning, and social support. AB - Thirty pairs of fathers and mothers who had school-age children with mental retardation and other disabilities were compared with each other and with 32 father and mother pairs of parents of children without disabilities. Responses to family scales indicated that fathers and mothers of children with developmental disabilities did not differ from each other nor from fathers and mothers of children without disabilities in parental stress, family social support, or family functioning. However, parents of children with disabilities experienced a disproportionately greater level of stress relating to their children than did those of children without disabilities. Fathers' and mothers' stress was associated with aspects of family functioning as perceived by themselves and their spouses. PMID- 9394134 TI - Disaggregating parental depression and family stress in assessing families of children with developmental disabilities: a multisample analysis. AB - The psychometric properties of a 5-item subcomponent (DEP5) of Factor 1, Parent and Family Problems, of the Friedrich Questionnaire on Resources and Stress were examined to determine whether these items formed a depression subcomponent of the larger family stress factor. Data were pooled from two samples numbering more than 450 respondents. Internal analyses established that the DEP5 had adequate internal reliability and stability over 2 years. Furthermore, confirmatory factor analyses indicated that the DEP5 measured depressive reactions as distinct from other parent and family problems. These results should be useful to researchers who want to reanalyze the Questionnaire on Resources and Stress data specifically for depressive reactions. Also, the technique for disaggregating global instruments can be applied to other variables. PMID- 9394136 TI - Reliability of ratings of consumers with mental retardation and their staff on multiple measures of social support. AB - Reliability of self-reports of social support with staff ratings was compared through determining the internal consistency of the measures, consistency across measures, and consistency across raters. Fifty adults with mild mental retardation and their staff in supported living residential settings were interviewed. Self-report ratings had moderate internal consistency, were consistent across rating scales, and were significantly correlated with staff ratings, although staff members tended to agree more with each other than with consumers. Results suggest that individuals with mild mental retardation can be reliable reporters about their own social support. Further, examining self informant agreement for specific support sources can illuminate discrepancies between self- and informant-obtained ratings. PMID- 9394137 TI - Generalized identity matching of two-dimensional forms by individuals with moderate to profound mental retardation. AB - The classic literature suggests that individuals with MAs of less than 5 years may fail tasks that require same/different judgments. In Study 1 we used an assessment procedure that provided minimal instructional programming to determine whether 17 adults with MAs ranging from 2 years, 5 months to 4 years, 11 months would show accurate identity matching-to-sample. Stimuli were letter-like nonsense forms. Eight participants showed highly accurate matching. Eight of the 9 who failed were available for further study. Of these, 5 ultimately demonstrated highly accurate matching after training with standard fading procedures. These data suggest that a greater proportion of individuals with low MAs can exhibit generalized identity matching than previously documented in the literature. PMID- 9394138 TI - Differences in social signals produced by children with developmental delays of differing etiologies. AB - Clarity of referential looks (either a focus on parent's face or other focus) produced by preschool children with delays of differing etiologies and children without delays was examined. Adults (with and without experience with children with delays) viewed videotaped segments in which children's looks did or did not occur. Adults judged whether a look occurred and rated their confidence in each judgment; latency to respond was measured. Adults' experience with children with delays did not influence outcome measures. When viewing looks focusing on parents' faces, participants were more accurate and more confident judging looks by children with typical development, less accurate when viewing face-directed looks of children with developmental delays, and least accurate when viewing children with Down syndrome. Discriminability of social looks differed by etiological group, and judges' decision criteria, confidence, and speed of responding also differed. PMID- 9394139 TI - Therapist-child interaction in the middle minutes of sensory integration treatment. AB - The purpose of this study was to describe the management of challenge during therapist-child interaction in sensory integration treatment. This descriptive and relational study of the middle minutes of treatment sessions partially replicated an earlier study of the beginning minutes. One-minute videotape clips taken from the middle minutes of 38 treatment sessions were shown to therapist judges who rated qualities of therapist and child behavior. Two patterns emerged from the correlations of ratings: work and playfulness. Work for the child involved trying hard, cooperating and seeking assistance, whereas work for the therapist involved assisting and guiding the child. Play for the child included enjoying the activity, being successful and confident, and trying hard. For the therapist, play involved being creative and behaving playfully. Patterns of work and play were different across different levels of challenge to the child. PMID- 9394140 TI - Clinical interpretation of "Therapist-child interaction in the middle minutes of sensory integration treatment". PMID- 9394141 TI - Symbolic play language during sensory integration treatment. AB - OBJECTIVE: Clinical writings on sensory integration treatment and theory have long professed that play serves as an important means of implementing treatment goals. However, to date, there has been little research that examines this aspect of the intervention. With the use of play language as an indicator for the occurrence of play, this study examined the frequency and characteristics associated with symbolic play language that therapists and children use during sensory integration therapy. This study is part of an ongoing research program designed to examine therapist-child interactions. METHOD: The frequency of symbolic play language observed in 41 videotaped treatment sessions of therapist child dyads (21 children, 12 therapists) was recorded with the Challenge Coding System. The presence of symbolic play language was recorded if the child or therapist used language that incorporated the child, therapist, equipment, or activity into a symbolic or pretend play theme. The frequency of symbolic play language and percentage of time spent using play language were calculated. Associations among frequency of play language, child age, and behavior during the session (e.g., seeking assistance, cooperation) were also examined. RESULTS: Symbolic play language proved to be a major feature of sensory integration treatment sessions. It also correlated with child age and with some features associated with therapeutic interactions (i.e., child tries hard, child seeks assistance, therapist assists child, therapist modifies activity, therapist structures activity). CONCLUSION: The results suggest that these therapists used play language frequently and that this usage may support children in sensory integrative therapy to successfully accomplish activities. PMID- 9394142 TI - The Interest Checklist: a factor analysis. AB - OBJECTIVE: The purpose of this study was to determine whether the 80 items on the Interest Checklist empirically cluster into the five categories of interests described by Matsutsuyu, the developer of the tool. METHOD: The Interest Checklist was administered to 367 subjects classified in three subgroups: students, working adults, and retired elderly persons. An 80-item correlation matrix was formed from the responses to the Interest Checklist for each subgroup and then used in a factor analysis model to identify the underlying structure or domains of interest. RESULTS: Results indicated that the Social Recreation theoretical category was empirically independent for all three subgroups; the Physical Sports and Cultural/Educational theoretical categories were empirically independent for only the college students and working adults; and the Manual Skills theoretical category was empirically independent for only the working adults. CONCLUSION: Although therapists should continue to be cautious in their interpretation of patients' Interest Checklist scores, the tool is useful for identifying patients' interests in order to choose meaningful activities for therapy. PMID- 9394143 TI - Forced use of the upper extremity in cerebral palsy: a single-case design. AB - OBJECTIVE: Muscle imbalance and poor control of movement can have an impact on the daily occupational functioning of children with cerebral palsy. When one side of the body functions better than the other, children will often prefer to use the less-involved upper extremity for completion of play and self-care activities because they have learned that the other hand does not function as effectively. This study examined a method purported to overcome this learned nonuse of the affected upper extremity by directing the child's attention to this extremity and increasing his or her motivation to use it. The research hypothesis was that restriction of the less-involved hand with a resting splint would result in increased use of the more-involved hand in a child with spastic cerebral palsy. METHOD: Initially, two children with cerebral palsy participated in this single subject, ABA design study, but only one subject complied with the splint-wearing schedule and completed the study. This subject was a 2-year-old girl with greater involvement of the right side than the left. During the experimental phase, she wore a resting splint on her less-involved hand for most of the waking hours of the day to restrict its use. Data were collected over a 7-week period (2 weeks presplinting, 3 weeks splinting, 2 weeks postsplinting) and at a 6-month follow up. Use of the more-involved extremity was measured through analysis of her performance during 15-min videotaped sessions of free play, administration of items from the Peabody Developmental Fine Motor Scales, and completion of a daily finger-feeding task. RESULTS: An improvement in quality, quantity, and variety of use of the more-involved extremity after splinting, with some continuing improvement evident at follow-up, was found. The subject had increased voluntary control of her more-involved arm and hand and used them more spontaneously for completion of daily occupations. CONCLUSIONS: Although the results of this single case design are encouraging, further research with a randomized, controlled design is necessary to determine the effectiveness of the forced-use technique with a larger population. PMID- 9394144 TI - Development of a post-offer screening tool for patient support services. AB - OBJECTIVES: The purpose of our project was to develop a post-offer screening tool that demonstrates interrater reliability, predictive validity, and face validity and that accurately represents the physical demands of the patient support services (lifting team) job at our health care facility. METHODS: The screening tool, which consists of 11 static and dynamic tasks, was developed, using the 13 incumbent staff members of the patient support services department, to determine whether the criteria established for each task matched the physical abilities of at least 80% of the total group tested. Test-retest design was used for this study. Intraclass correlation coefficients and the Kappa statistic were used to calculate interrater reliability. Face validity was determined through the Job Similarity Questionnaire completed by all subjects. RESULTS: Subjects did not meet criteria established for the static knee pull and the knuckle-to-elbow lift tasks, resulting in modification of these two criteria. Interrater reliability ranged from .22 for the maximum static pull wall task to .94 for the left-hand grip strength task. Face validity ranged from 53.9% to 92.4%. CONCLUSION: Although face validity of the Job Similarity Questionnaire represented a wide range, we believe that the results were homogeneous enough to continue with the screening tool unchanged, except for lowering the expected outcome on two tasks. Interrater reliability was established for 75% of the tasks. The lack of variation of data for the other 25% prevented statistical analysis of those tasks but confirmed that all members met the physical criteria. PMID- 9394145 TI - The use of service learning in client environments to enhance ethical reasoning in students. AB - OBJECTIVES: This study examined the effects of two service-learning experiences on the psychosocial and moral reasoning development of occupational therapy students. The assumption was that ethical reasoning ability can be facilitated through participation in value-laden experiences. METHOD: Participants visited older adults in nursing homes (n = 19) or interacted with persons with disabilities in community settings (n = 33). All participants reflected on their experiences through weekly journals. Psychosocial and moral reasoning development were measured at the beginning and end of the experiences with the Student Development Task and Lifestyle Inventory and the Sociomoral Reflection Measure Short Form. RESULTS: Participants in both groups exhibited a significant time related increase in psychosocial development but no increase in moral reasoning. Participants interacting with persons with disabilities exhibited a decrease over time in moral reasoning compared with the participants interacting with older adults. CONCLUSION: Service learning effected a change in the participants' psychosocial development indicative of developing an appreciation for dignity, equality, and justice. These are core concepts in occupational therapy and are viewed as important in ethical reasoning. The lack of advancement and current level of moral reasoning in these undergraduate students raises a question as to their readiness to engage in ethical reasoning as entry-level practitioners. PMID- 9394146 TI - Cross-training concept paper. PMID- 9394147 TI - Statement--sensory integration evaluation and intervention in school-based occupational therapy. AB - Based on the educational team recommendations, school-based occupational therapy practitioners provide interventions to students who are eligible for special education services under IDEA and who need occupational therapy to benefit from their education program. The occupational therapist develops an intervention plan based on the student's needs and the therapist's professional knowledge base. When students demonstrate deficits in sensorimotor performance components that contribute to a significant and documented discrepancy in their skills within their educational program, the use of sensory integrative approach may be one frame of reference for intervention chosen by the occupational therapist. PMID- 9394148 TI - Statement--fundamental concepts of occupational therapy: occupation, purposeful activity, and function. AB - Deriving from the philosophical basis of the profession, occupation is the core concept of the profession of occupational therapy. However, in the occupational therapy literature, the term occupation is used in a variety of ways. Occupation, a collection of activities that people use to fill their time and give life meaning, is organized around roles or in terms of activities of daily living, work and productive activities, or pleasure, for survival, for necessity, and for their personal meaning. It is the individualized, unique combination of activities that comprises an individual's occupations. Purposeful activities have been described in many different ways: as something all people engage in; as tools or media that therapists use to enhance or facilitate performance; and vehicles for bringing about change. Purposeful activities are seen as part of the process of occupational therapy. Purposeful activities are subset of occupations in that they are goal directed and serve as a major tool in the process of occupational therapy. The term function, viewed as the ability to perform activities required in one's occupations has become increasingly important to society in describing the performance or change in individuals. This societal shift in ideas has prompted the concept of function, viewed as a product, to become more important than the process of bringing about change. Occupational therapy practitioners typically have viewed the process as being just as important as the product. When working to improve function, occupational therapy practitioners use purposeful activities that are meaningful to the person in relation to his or her occupational history, preferences, personal goals, and needs. Occupational therapy practitioners need to keep the individual's occupations in the forefront of their thoughts when using any purposeful activity and to plan interventions toward improving the individual's ability to function within his or her occupations. In the interest of the profession, it is important to concentrate on occupation. Furthermore, it is essential that we study our interventions' relationship to occupation and function, and how purposeful activities are used toward supporting the individual's ability to engage in occupation. PMID- 9394150 TI - Physical agent modalities position paper. PMID- 9394149 TI - The psychosocial core of occupational therapy position paper. PMID- 9394151 TI - Occupational therapy's link to vocational reeducation, 1910-1925. AB - The development of occupational therapy is rooted in early 20th century medical reform. During the early 1910s, several members of the medical profession, human service workers, and the larger American society were increasingly disturbed by medical practices that did not consider the individual's personal experience of disability. Occupational therapy was developed, in part, out of this desire to provide persons with treatment that helped them to function in their communities despite their disability. Early occupational therapy leaders envisioned the fledgling profession as a societal service capable of assisting persons with disabilities to return to both work and community life. Vocational reeducation was initially considered to be an integral component of occupational therapy in the years from 1910 to 1920. However, the profession's early link to vocational reduction was challenged by vocational technical trainers during World War I. To prevent occupational therapy from being subsumed by vocational technical training, the early occupational therapy leaders implemented several strategies: adoption of physician prescription for all occupational therapy services, delivery of occupational therapy services primarily within hospital settings, and dissociation from vocational reeducation services. Reasons accounting for why the early occupational therapy leaders abandoned their initial commitment to vocational reeducation are explored. Suggestions about how this decision has affected present-day practice are also offered. PMID- 9394152 TI - Identifying best practice in the occupational therapy assistive technology evaluation: an analysis of three focus groups. PMID- 9394153 TI - Positioners for wheelchairs in long-term-care facilities. PMID- 9394155 TI - Impact of promotion of occupational therapy campaign. PMID- 9394156 TI - Professional values: diversity versus disintegration. PMID- 9394154 TI - Spirituality issue provides needed voice. PMID- 9394157 TI - Reversible oligohydramnios in a pregnancy with angiotensin-converting enzyme inhibitor exposure. AB - The use of angiotensin-converting enzyme inhibitors during pregnancy has been associated with poor fetal outcomes, including oligohydramnios, renal tubular dysplasia, cranial malformations, and fetal death. A 35-year-old woman with chronic hypertension was treated with the angiotensin-converting enzyme inhibitor benazepril until 27 weeks' gestation, when severe oligohydramnios was noted. After hospitalization for bed rest, fetal surveillance, and discontinuation of the agent, amniotic fluid rapidly reaccumulated, and a healthy infant was delivered at term. Although the use of angiotensin-converting enzyme inhibitors should be avoided during pregnancy, patients whose fetuses are inadvertently exposed in utero need not be given a uniformly poor prognosis. Oligohydramnios induced by the use of angiotensin-converting enzyme inhibitors during pregnancy may be reversible if the agent is discontinued. This case underscores the need for obstetricians to review carefully the medication regimens of all pregnant women and to be familiar with generic and proprietary names of medications to avoid the use of potentially harmful agents during pregnancy. PMID- 9394158 TI - Relative humidity under radiant warmers: influence of humidifier and ambient relative humidity. AB - The effects of humidifier air temperature and flow, and ambient relative humidity (RH(amb)) on RH and air temperature under a radiant warmer (RW) were determined in stable and unstable conditions, using an infant surrogate. Mean supplemented RH under the RW was 36.3% at 14% RH(amb) and 67.6% at 55% RH(amb). Humidifier temperature of 38 degrees C and air flow of 10 LPM produced highest RHs (74.5% and 43.1% in high and low RH(amb), respectively). RH(amb) was highest in summer and lowest in winter in this midwest U.S. hospital, and could be predicted by calendar date (r = 0.58). Humidification equipment capabilities and limitations must be known when using this method to limit evaporative water loss. PMID- 9394159 TI - Ilio-psoas abscess in a neonate. AB - A full-term, small-for-gestational-age, neonate was born 4 days after rupture of the membranes. On the 5th day of life, she developed sepsis due to Klebsiella pneumoniae. On the 18th day of life, the right hip was noted swollen with limited range of motion, but it was painless on passive movements. Ultrasonography revealed abscess of the right ilio-psoas muscle with normal appearance of the right hip joint. Surgical incision and drainage and antibiotic administration resulted in a gradual full recovery. Ultrasonography can confirm the diagnosis of this exceptional clinical entity in neonates, which is difficult to differentiate from septic arthritis of the hip. PMID- 9394160 TI - The risk of neonatal death and respiratory distress syndrome in relation to birth weight of preterm infants. AB - The aim of this study was to determine the risk of respiratory distress syndrome (RDS) and neonatal death (NND) in relation to birth weight in preterm neonates. A group of 255 singleton preterm neonates born at 22-36 weeks were examined. The mean birth weight for each gestational week was estimated from a fitted curve. Each birth weight was recalculated as a multiple of the mean. This approach allowed description of a range of birth weight for the whole population of preterm infants. As expected, the incidence of neonatal death and respiratory distress syndrome was higher among the less mature infants. However, there were no significant differences in outcome (NND, severe RDS) between infants whose birth weight was above or below the mean for gestational age, and no excess of "growth retardation" (birth weight below the 5th centile) in babies with NND or RDS. We conclude that, in our population, the risk of RDS and neonatal death does not appear to be related to the birth weight of preterm neonates but is, of course, related to gestational age. PMID- 9394161 TI - Vaginal delivery is associated with occult disruption of the anal sphincter mechanism. AB - Childbirth is thought to be an important cause of pelvic floor dysfunction. Heretofore, this has been thought due to pudendal denervation. Endovaginal sonography allows thorough assessment of the anorectum and in this study was used to assess nulliparous women and women before and after delivery. Two groups were studied. Thirty-two nulliparous subjects without complaints of incontinence were studied once. Thirty-four pregnant women were studied before and after delivery. Endovaginal sonography was used to assess integrity of internal and external anal sphincters, thickness of the levator bundle, internal and external sphincters, anal length, and the angle between the levator bundles. Delivery was associated with disruption of the internal and external sphincters. No nulliparous women (nonpregnant or pregnant) had sphincter disruption demonstrated. Episiotomy in the index delivery was associated with increased thickness in the external sphincter and a smaller angle between the levator bundles. Vaginal delivery is associated with occult disruption of the anal sphincters. PMID- 9394162 TI - The effects of essential fatty acids preparation in the treatment of intrauterine growth retardation. AB - A treatment of intravenous infusion of glucose, amino acids, and emulsion enriched in essential fatty acids (EFAs), linoleic, and linolenic acids were given to 30 pregnant women with intrauterine growth retardation (IUGR) and 28 non EFAs treated cases as controls. There was a marked gain in fetal biparietal diameter (BPD) and in the estimated weight of the treated group over the control group. The mean birth weight was significantly different in the two groups. Fetal BPD increased much more in patients treated with EFAs at 28-34 gestational weeks than those at 34 1/7-37 weeks, which indicates that early initiate complement of n-3 and n-6 fatty acids to IUGR mothers may correct pregnancy-induced EFAs deficiency and maternal-fetal malnutrition, which demonstrates a fetal catch-up of growth in the brain and the whole body. PMID- 9394163 TI - Growth and growth factors in premature infants receiving dexamethasone for bronchopulmonary dysplasia. AB - Physical growth and the serum growth factors, insulin growth factor 1 (IGF1) and its binding protein (IGFBP3) were measured weekly during dexamethasone treatment and for 3 weeks after stopping therapy in 10 ventilated babies [median (range) birth weight 860 g (640-1210); median (range) gestational age 26 weeks (24-29)] with bronchopulmonary dysplasia (BPD). The mean (+/- SE) rates of change of all physical measures except crown-rump length (CRL) increased significantly after stopping dexamethasone: weight gain 13.2 (+/- 1.5) on versus 1.0 (+/- 1.9) g/day off treatment; occipital-frontal circumference 0.7 (+/- 0.1) cm/week; CRL 0.5 (+/ 0.1) versus 0.7 (+/- 0.1) (TBL) 0.7 (+/- 0.1) versus 1.1 (+/- 0.1) cm/week; CRL 0.5 (+/- 0.1) versus 0.7 (+/- 0.1) cm/week, and knee-ankle length (KAL) 0.13 (+/- 0.02) versus 0.36 (+/- 0.04) cm/week. Mean serum IGF-1 (1.57 +/- 0.13 versus 3.56 +/- 0.41 nmol/L) and IGFBP3 (0.94 +/- 0.03 versus 1.12 +/- 0.05 mg/L) levels also increased off treatment. The weekly dose of dexamethasone (mg/kg) was significantly negatively correlated with all physical growth measures (P < 0.01), but showed no correlation with growth factors. Protein intake (g/kg/day) was significantly correlated (P < 0.01) with weight gain (r = 0.28), changes (TBL) (r = 0.32), serum IGF1 levels (r = 0.60), and IGFBP3 levels (r = 0.37). All aspects of physical growth are compromised during dexamethasone treatment for BPD. Poor growth during steroid treatment is associated with lower IGF1 and IGFBP3 levels. Further study is needed to examine the effect of varying dexamethasone dosage regimes and nutritional intake on the growth process in BPD. PMID- 9394164 TI - The effect of maternal glycemic control on fetal growth in diabetic pregnancies. AB - In this prospective study, we examined the effect of maternal glycemic control on fetal growth in pregnancies complicated by pregestational diabetes. One hundred and sixty-five pregestational diabetic pregnancies were studied with serial ultrasound scans and fetal growth was examined as a function of maternal glycemic control. There was a significant, although small, reduction in fetal biparietal diameter growth rate in the presence of poor maternal glycemic control during the first half of the pregnancy. In the second half of pregnancy, maternal hyperglycemia contributed to fetal macrosomia. We conclude that in pregnancies with pregestational diabetes, maternal hyperglycemia affects fetal growth in a biphasic manner. As a result of that, although babies born to diabetic mothers appear of relatively overall normal size and weight, they may have smaller heads than their potential and more fat. PMID- 9394165 TI - Penicillin desensitization in the treatment of syphilis during pregnancy. AB - The objective of this study was to compare patients' hospital course, complications, and charges for oral and intravenous (i.v.) desensitization regimens for the treatment of syphilis in the penicillin-allergic gravida. We performed a retrospective search of medical records at two tertiary-level teaching hospitals and reviewed the hospital course of penicillin-allergic gravidas who underwent penicillin desensitization. Between August 1988 and December 1995, 16 procedures for penicillin desensitization were carried out: 11 oral procedures, and 6 i.v. procedures. There were no significant differences between the patients in the oral and i.v. desensitization groups with respect to demographic characteristics, duration of time in a monitored bed, or length of hospital stay. The oral regimen was less expensive than the i.v. regimen ($144.06 vs. $319.48). In our experience, oral and i.v. regimens provide effective desensitization for the treatment of syphilis in penicillin-allergic gravidas. However, the oral route offers ease of administration and substantial cost savings, making it the preferred method. PMID- 9394166 TI - Nuchal cord entanglements and gestational age. AB - The purpose of this observational study was to investigate the relation between nuchal cord entanglements and gestational age. Computerized data from our hospital perinatal database were reviewed between January 1990 and December 1994. Data from all deliveries > or = 20 weeks' gestation underwent either a Cochran Mantel test for trend or Chi-square testing where appropriate. Of the 13,895 singleton deliveries, the finding of an entanglement increased significantly from 5.8% at 20 weeks to 29.0% at 42 weeks' gestation. The frequencies increased linearly, regardless of whether the entanglement involved a single loop (p < 0.001) or multiple loops (p < 0.002). The risk of an antepartum stillbirth was not increased in the presence of a nuchal cord entanglement, even after controlling for other risk factors. These normative data should serve as a reference for prenatal ultrasound examinations and for prospective studies intended to evaluate the predictive value of reporting this finding prenatally. PMID- 9394168 TI - Fetal acidemia in the absence of fluid observed at amniotomy. AB - The objective of this study was to determine the rate of pathological fetal acidemia in the absence of fluid observed at amniotomy. Thirty-nine consecutive patients with no fluid observed at the time of amniotomy were prospectively enrolled in this study. Ultrasound measurement of amniotic fluid index was performed. Umbilical cord gases were performed on arterial and venous samples at the time of delivery. Patient name and medical record number were noted and delivery data were extracted from review of the medical record. The median gestational age at admission was 41 weeks (range 38 to 42 weeks). Sixteen patients (41%) were subsequently noted to have meconium at the time of delivery. The median amniotic fluid index was 2.0 cm with a range of 0 to 9.0 cm. Thirty patients (76.9%) had an amniotic fluid index of less than 5.0 cm. The median umbilical artery pH in this patient population was 7.21 with a range of 6.75 to 7.42. Only one infant had an umbilical artery pH less than 7.00. The rate of cesarean section for documented fetal distress was 2.6%. The absence of observed fluid at amniotomy, while commonly associated with subsequent meconium at delivery, is not predictive of fetal acidemia or operative delivery for fetal distress. PMID- 9394167 TI - Urinary cyclic GMP, endothelin, and prostaglandin E2 in normal pregnancy and preeclampsia. AB - Cyclic GMP, endothelin and prostaglandin E2 (PGE2) all have systemic vasoactive properties (with cyclic GMP acting as a second messenger of nitric oxide). Intrarenally they act as natriuretics and urinary levels reflect intrarenal production. Cyclic GMP and PGE2 also act as important inhibitors of platelet activation and thrombosis. The purpose of this study was to determine if urinary levels of cyclic GMP, endothelin, and PGE2 differ in preeclamptic as compared to normal pregnancies. Parameters were compared in 13 normotensive, nonpreeclamptic pregnancies, and 32 preeclamptic pregnancies. Preeclamptic women had significantly lower levels of urinary cyclic GMP (0.67 +/- 0.12 vs. 2.1 +/- 0.5 nmol/g creatinine), endothelin (0.88 +/- 0.09 vs. 3.75 +/- 1.4 ng/g creatinine), and PGE2 (26 +/- 4 vs. 9 ng/g creatinine) as compared to normals (p < 0.05). Intrarenal production of cyclic GMP, endothelin, and PGE2 are all disturbed in preeclampsia and may have implications in the sodium retention, hypertension, and intrarenal thrombosis and vasospasm of preeclamptic pregnancy. PMID- 9394169 TI - The Cantrell-sequence: a result of maternal exposure to aminopropionitriles? AB - The characteristic features of the Cantrell-sequence--anterior thoraco-abdominal wall defect with ectopia cordis and diaphragm, sternum, pericardium, and heart defects--have been observed in animals following maternal administration of beta aminopropionitrile, a toxic amino-acid derivative. We report on an unusual case of the Cantrell-sequence in a premature infant with associated dysmelia, aplasia of the right kidney, cerebellar hypoplasia and circumscribed aplasia of the cutis, which has not been reported previously. Maternal history suggested an occupational exposure to aminopropionitriles prior to pregnancy. Prenatal ultrasound, differential diagnosis, perinatal management, and the teratogenic role of aminopropionitriles in this rare genetic disorder are discussed. PMID- 9394170 TI - Liposomal amphotericin B in neonates with invasive candidiasis. AB - Liposomal amphotericin B L-Amp B, a novel formulation of Amp B, is effective for the treatment of invasive fungal infections in children and adults and is associated with less toxicity than the conventional preparation. Data on the use of L-Amp B in neonates is scarce. We describe the clinical course of two premature infants who were treated with L-Amp B (one infant had candidemia, and the other had candidemia and meningitis), and provide a summary of previously published experience on this topic. L-Amp B may be an option for therapy of invasive candidiasis in neonates who are at high risk of nephrotoxicity and other amphotericin-related reactions, but clinical trials are necessary to document its safety and efficacy in this age group. PMID- 9394171 TI - Early-onset neonatal sepsis in Pakistan: a case control study of risk factors in a birth cohort. AB - We prospectively evaluated risk factors for early-onset neonatal (EON) sepsis in a case-control study among inborn patients at the Aga Khan University Medical Centre in Karachi between 1990-1993. A total of 38 cases with blood culture proven bacterial sepsis were identified within 72 hr of birth (prevalence 5.6 of 1000 live births) and matched with two consecutive gender matched births with no complications. The most common isolates were Staphylococcus aureus (18%), group B Streptococci (13%), and Klebsiella pneumoniae (13%). Univariate analysis of maternal risk factors revealed a significant association between maternal urinary tract infection (UTI) (odds ratio [OR]20, 95% confidence interval [CI]2.4-166.9), maternal pyrexia (P < 0.0001), vaginal discharge (P < 0.05), vaginal examinations during labor (P = 0.03), and EON sepsis. The infected newborns also had significantly lower apgar scores at birth (P < 0.0001) and a significantly greater number were intubated at birth (Fisher's exact test P = 0.04). Infected newborn infants were transferred out of the labor room earlier than noninfected controls and significantly fewer received exclusive breastfeeds (OR 0.33, 95% CI 0.1-0.8). Our data suggest the possibility that both vertical transmission from the mother as well as postnatal acquisition of infection from the environment may be of importance in the pathogenesis of EON sepsis in Karachi. Preventive measures should focus at recognition of high-risk infants, strict asepsis during labor, and early institution of exclusive breastfeeding. PMID- 9394172 TI - Exposure to pulsed magnetic fields in the treatment of plantar ulcers in leprosy patients--a pilot, randomized, double-blind, controlled clinical trial. AB - A pilot, randomized, double-blind, controlled clinical trial to study the effect of exposure to pulsed magnetic fields (PMF) on the rate of healing of plantar ulcers in leprosy patients was undertaken. Twenty patients were randomly allocated to receive standard wound-care treatment (controls) and 20 others received standard treatment plus exposure to PMF (sinusoidal form, 0.95 to 1.05 Hz, amplitude +/- 2400 nano Teslas) (study group) for four weeks. Assessment of the outcome of treatment was based on the volume of ulcers, calculated from the maximal length, breadth and depth of the ulcer recorded on the day of admission, at one and two weeks and at the end of treatment. The analysis of the results was based on 15 control patients and 18 PMF patients after deletion of four patients due to irregularity in attendance and three others on account of suspected malignancy of the ulcers. In the control group, the geometric mean volumes of the ulcers were 2843 and 1478 cu mm on the day of admission and at the end of the treatment (P = 0.03); the corresponding values in the PMF group were 2428 and 337 cu mm, respectively (P < 0.001). A decrease in the volume of 40% or more was observed in 53% of control patients and 89% of PMF patients (P = 0.02); a decrease of 80% or more was observed in none of the controls and in 33% of PMF patients. These findings strongly suggest that exposure to PMF causes a significantly more rapid healing of plantar ulcers in leprosy patients. PMID- 9394173 TI - Semen biochemistry of leprosy patients. AB - Studies have been made on the semen of three categories (borderline, borderline tuberculoid and lepromatous) of leprosy patients to evaluate the seminal biochemical constituents viz. fructose, glycerylphosphorylcholine and acid phosphatase besides the physical properties viz. volume, pH, liquefaction time, sperm density and sperm motility. In all categories of leprosy patients, seminal pH, liquefaction time and sperm density underwent significant decline. The decline in the seminal volume and sperm motility was significant only in borderline leprosy. It was observed that seminal glycerylphosphorylcholine (GPC) concentration and acid phosphatase activity declined in all categories of leprosy patients but GPC showed a significant decline only in borderline tuberculoid and acid phosphatase declined significantly only in borderline and lepromatous leprosy. PMID- 9394174 TI - Profile of leprosy in children: past and present. AB - The profile of leprosy in children currently seen in a referral hospital is compared with that of children with leprosy admitted in the 1970s. Children with leprosy under the age of 15 years in 1974 and 1979 comprised one group (Group I) while those during 1989 and 1994 constituted the second group (Group II) The variables studied included age, sex, type of leprosy, deformity and contact status. Multidrug therapy (MDT) was introduced in the treatment of leprosy in 1982. The probable change it has made in the presentation of leprosy in children is discussed. PMID- 9394175 TI - Concurrent leprosy and HIV infection: a report of three cases. AB - Three cases of concurrent infection with HIV and leprosy are reported. One had developed borderline lepromatous leprosy one year after identifying HIV infection, while the other two had indeterminate leprosy and both conditions were identified at the same time in these two patients. All three cases showed satisfactory response to standard antileprosy multidrug therapy. PMID- 9394176 TI - Shortening duration of treatment of multibacillary leprosy. Action Programme for the Elimination of Leprosy, WHO. PMID- 9394177 TI - Hidden cases of leprosy (in prison) AB - Leprosy survey conducted in eight prisons in seven districts of Bihar State revealed a prevalence of 13.3 per 1000 which was 12 times more than the recorded prevalence of leprosy in the State. Thus this finding supports the view that prisons could form a hyperendemic pocket for leprosy. Regular NLEP services need to be extended to the inmates of the prisons. PMID- 9394178 TI - Cauda equina syndrome masquerading as leprosy. PMID- 9394179 TI - Profundus minus deformity in leprosy. PMID- 9394180 TI - Building bridges and controlling parasites. PMID- 9394181 TI - Australasian contributions to anti-parasite vaccines. PMID- 9394182 TI - The Bancroft-Mackerras Oration. Livestock parasite treatment--a call for greater interaction between research and industry sectors. PMID- 9394183 TI - Livestock parasite treatment--a call for greater interaction between research and industry sectors. AB - During the past 2 decades, treatment of livestock parasitism has changed from a situation of relative stability, particularly in the mid-late 1980s when the macrocyclic lactone compounds were introduced, to the situation in many countries now where control of sheep parasites is rapidly being lost. As serious as the situation may be, now is not a time to wring our hands in despair but a time to address some of the limitations and identify opportunities where a greater integration of the efforts of research and industrial bodies might be directed to maintaining parasite control in our livestock industry. PMID- 9394184 TI - Molecular methods for diagnosis and epidemiological studies of parasitic infections. AB - Direct microscopy is widely used for the diagnosis of parasitic infections although it often requires an experienced microscopist for accurate diagnosis, is labour intensive and not very sensitive. In order to overcome some of these shortcomings, molecular or nucleic acid-based diagnostic methods for parasitic infections have been developed over the past 12 years. The parasites which have been studied with these techniques include the human Plasmodia, Leishmania, the trypanosomes, Toxoplasma gondii, Entamoeba histolytica, Giardia, Trichomonas vaginalis, Cryptosporidium parvum, Taenia, Echinococcus, Brugia malayi, Wuchereria bancrofti, Loa loa and Onchocerca volvulus. Early methods, which involved hybridisation of specific probes (radiolabelled and non-radiolabelled) to target deoxyribonucleic acid (DNA), have been replaced by more sensitive polymerase chain reaction (PCR)-based assays. Other methods, such as PCR hybridisation assays, PCR-restriction fragment length polymorphism (PCR-RFLP) assays and random amplified polymorphic DNA (RAPD) analysis have also proved valuable for epidemiological studies of parasites. The general principles and development of DNA-based methods for diagnosis and epidemiological studies will be described, with particular reference to malaria. These methods will probably not replace current methods for routine diagnosis of parasitic infections in developing countries where parasitic diseases are endemic, due to high costs. However, they will be extremely useful for genotyping parasite strains and vectors, and for accurate parasite detection in both humans and vectors during epidemiological studies. PMID- 9394185 TI - Designer vaccines for parasitic diseases. AB - Conventional ways of developing vaccines against infections, either on pragmatic grounds or by identifying protective antigens and attempting to mimic natural immune responses, have largely been unsuccessful for parasitic-infections, mainly because of the complexity of the immunological processes involved. It is clear that a new approach is required and it is now known that the "immunological environment" in which the immune response is initiated is as, or more, important than the actual antigens used. CD4+ and CD8+ T1 cells, through the agency of IL-2 and IFN-gamma, direct the response towards cell-mediated immunity involving cytotoxicity and macrophage activation, whereas T2 cells, through the agency of IL-4 and IL-10, direct the response towards antibody production. The two poles are counter-regulatory in that IFN-gamma inhibits antibody formation and IL-4 and IL-10 inhibit macrophage activation. However, immune responses are not immutable and can be artificially driven towards one or other pole, for example IFN-gamma, IL-2 and IL-12 favour T1 responses, whereas IL-4 and IL-10 favour the T2 type. With this knowledge, it is possible to design recombinant or nucleic acid vaccines that include gene products or genes for desirable cytokines as well as the appropriate antigen. For example, in experimental leishmaniasis, protective immune responses can be induced by the incorporation of genes for IL-2 and IFN gamma into recombinant Salmonella typhimurium vectors and nucleic acid vaccines. A similar approach might be appropriate in experimental schistosomiasis, in which exogenous IL-12 drives the immune response towards the T1 pole and ameliorates T2 mediated pathology. These approaches require novel delivery systems and these have already begun to produce encouraging results. However, simply modifying the nature and route of administration of the vaccine is not enough and attention has now turned to the effector molecules involved, for example nitric oxide, and the signaling systems that are modified by the presence of particular cytokines. PMID- 9394186 TI - The importance of transmission-blocking immunity in the control of infections by apicomplexan parasites. AB - Transmission-blocking immunity may have great potential for use in the control of diseases caused by apicomplexan parasites. In this review I will describe our work on the application of transmission-blocking immunity to the control of the Eimeria parasite and compare our results to those working on transmission blocking immunity against Cryptosporidium and Plasmodium. Eimeria causes the disease known as coccidiosis in domestic animals. Coccidiosis is particularly problematic in the chicken industry, mainly due to the crowded rearing conditions under which chicks are raised. In our work we identified, isolated and characterized 3 major gametocyte antigens (230 kDa, 82 kDa and 56/54 kDa) of Eimeria maxima. We used these native glycoproteins to immunize laying hens that, via the egg yolk, provide large amounts of transmission-blocking maternal antibodies to offspring chicks. We demonstrated that hatchlings from immunized hens shed 60-80% fewer oocysts (i.e. the infective stage of the life-cycle of Eimeria) than those from control hens. Such a reduction in oocyst output acts to significantly reduce parasite numbers in the litter of chicks raised in floor pens. This reduction in oocyst output is comparable to that seen using the most effective coccidiostat drugs and is probably sufficient to control coccidiosis under field conditions. Based on our results together with those of other groups working on transmission-blocking immunity against Cryptosporidium and Plasmodium, it appears that this immunological approach holds great promise for the control of apicomplexan parasites that cause diseases in both animals and man. PMID- 9394187 TI - Research collaboration in parasitology between Indonesia and Australia. AB - Indonesia and Australia are close neighbours sharing agro-ecological zones and common parasitological interests. Australia is an industrialised country and Indonesia is both industrialising and a developing country. The types of collaboration, contractual, collegiate, research collaboration and partnerships are briefly described. All forms of collaboration have and continue to exist between Australia and Indonesia. A survey of mammalian parasitology publications over the last 23 years indicates that the bulk of papers have been by Indonesian and non-Australian authors. Australian and Indonesian authors provided 4% of the total number of publications. The rational for collaboration is suggested to be the high degree of common multiple interests and the synergy of effort that can be derived from research partnerships. The most difficult issues in research collaboration are establishing the research priorities and, to a lesser extent, funding. The globalisation of the international research centre, International Livestock Research Institute, to include Asia will expand the opportunities for research collaboration. Details of the Australian Centre for International Agricultural Research mandate in supporting parasitology research collaboration is briefly described. The past and current research collaborative activities are reviewed and opportunities for future collaboration are listed. PMID- 9394188 TI - Control of parasites in cultured marine finfishes in Southeast Asia--an overview. AB - Mariculture in Southeast Asia began in the 1970s and expanded rapidly during the 1980s, with the commercial hatchery production of the seabass Lates calcarifer. Other important cultured species were Epinephelus coioides, Epinephelus malabaricus, Lutjanus johni, and Lutjanus argentimaculatus. Intensification in the polyculture of these species and the large-scale international movement of fingerlings or juveniles, as well as the rapid expansion and concentration of fish farms, have caused severe problems resulting from parasitic infections. Infections in maricultured fish are predominantly caused by monoxenous parasites, in particular the capsalid and diplectanid monogeneans. Heteroxenous blood parasites also successfully maintained transmission in the culture system despite their requirement for an intermediate host. Prophylactic chemical treatments helped to reduce parasitic infection but did not eliminate them and once introduced into the floating netcage culture system, these parasites managed to maintain their transmission successfully. Despite the current lack of information regarding the biology of many parasites affecting cultured marine fishes, it nevertheless is possible to develop methodologies to produce an integrated health management system specifically designed to the needs of the mariculture practiced in the Southeast Asian region. This system is important and should include a sequence of prophylaxes, adequate nutrition, sanitation, immunization and an effective system of marketing for farmed fishes. PMID- 9394189 TI - Control of freshwater fish parasites: a Southeast Asian perspective. AB - Parasites commonly found in freshwater fishes and other aquatic animals primarily belong to Protozoa, Platyhelminthes, Acanthocephala, Nematoda, Hirudinea and Crustacea. Most of these parasites are external parasites of the skin, fins or gills, while few are internal parasites living in the epidermal intralamellar gills, intestine or intramuscular tissues. Possible control measures involving manual removal, cleaning by topical cleaners, management practices, nutritional improvement, vaccination, chemoprophylaxis, chemotherapy, and quarantine and certification, etc., are discussed. PMID- 9394190 TI - Nematode parasite control of livestock in the tropics/subtropics: the need for novel approaches. AB - Because parasites are more abundant, small ruminants in the tropical/subtropical regions of the world experience much greater ravages from internal parasitic disease than those in the temperate regions. In the tropics/subtropics, the limiting ecological factor influencing the severity of parasitism is rainfall, as temperatures almost always favour hatching and development of the free-living stages. Attempts to expand sheep and goat production by replacing traditional village production systems, which rarely involve anthelmintic treatment, with large-scale intensive commercial enterprises invariably induce complete reliance on anthelmintics to control nematode parasites. This has led to the widespread development of high level, multiple anthelmintic resistance throughout the tropics/subtropics, and in certain regions this has reached the ultimate disastrous scenario of total chemotherapeutic failure. Immediate concerted efforts are needed to resolve this crisis. Significant benefits are likely to emerge from research into non-chemotherapeutic approaches to nematode parasite control, such as grazing management, worm vaccines, breed selection and biological control. However, it is likely that none, in isolation or collectively, will completely replace the need for effective anthelmintics. What is needed is the integration of all methods of parasite control as they come to hand, with the underlying aim of reducing the use and thus preserving the effectiveness of anthelmintics. Although cheap and simple procedures, based on sound epidemiological principles, can achieve dramatic benefits in worm control, they have been poorly adopted by livestock owners. Clearly then, the greatest need is for technology transfer and education programmes, but these activities are generally found to be chronically under-resourced. PMID- 9394191 TI - Basic and applied immunology in cestode infections: from Hymenolepis to Taenia and Echinococcus. AB - In larval cestode infections, it is well established that the intermediate mammalian host infected with egg-derived metacestodes in the tissue becomes completely immune to reinfection with eggs, whereas autoinfection has been conceived to occur in Hymenolepis nana/mouse (and human) and Taenia solium/human systems when these hosts are initially infected with metacestode-derived adult tapeworms in the lumen. In this review paper, the first topic is immunobiology of H. nana/mouse system on the reinfection immunity in order to get critical information as to how the initially ingested parasite (eggs or metacestodes) can develop into adult worms and how autoinfection does or does not occur in immunocompetent mice, since H. nana can complete its whole life cycle in the mouse intestinal tissue and lumen. When mice are infected with eggs (= oncospheres) of H. nana, they become immune to challenge infections with eggs within a few days (early response) and with cysticercoids within two weeks (late response). The initially established adult worms are expelled later (worm expulsion response). When mice are infected with cysticercoids, either derived from beetles or mice, they become immune to challenge infection with cysticercoids but not with eggs. Therefore, autoinfection occurs in the intestinal tissue for the establishment of cysticercoids in the tissue but never occurs in the intestinal lumen for the establishment of adult worms in immunocompetent mice. The second topic is vaccination trial against challenge infection with eggs of Asian Taenia in pigs. Pigs vaccinated with frozen oncospheres of Asian Taenia from Taiwan or Korea or T. saginata showed very strong resistance, whereas pigs vaccinated with those of T. solium showed partial resistance only. It is suggested that Asian Taenia is much closer to T. saginata than T. solium from the immunobiological viewpoint. The third topic is immunodiagnosis of echinococcosis and cysticercosis. Immunoblot analysis has revealed that Em18 (18 kDa component of crude antigens of Echinococcus multilocularis protoscolex) and glycoproteins of T. solium cysticerci are highly specific or unique to alveolar echinococcosis and cysticercosis, respectively. The fourth topic is discussion on miscellaneous prospects including laboratory animal models for echinococcosis and cysticercosis. PMID- 9394192 TI - A vaccine against the Asian schistosome, Schistosoma japonicum: an update on paramyosin as a target of protective immunity. AB - Paramyosin from parasitic worms of the genus Schistosoma has shown promise as a vaccine target and it is one of the candidates selected by WHO for the development of a vaccine against schistosomiasis. Here we discuss the literature of the past decade and report on different recombinant paramyosin constructs we are using in our laboratory to develop a vaccine against the Asian schistosoma, Schistosoma japonicum. PMID- 9394193 TI - Immunological approaches for the control of fasciolosis. AB - The immunological relationship between liver flukes and their mammalian hosts is being unravelled by in vivo and in vitro studies. Vaccine studies in cattle and sheep with purified antigens (fatty acid binding protein, FABP; glutathione S transferase, GST; cathepsin L, CatL; hemoglobin) have shown that high reductions in worm burdens (31-72%) and egg production (69-98%) can be achieved, raising the realistic possibility that immunological control of Fasciola infection is a commercially achievable goal. Combination vaccines may also be feasible since a cocktail of CatL and hemoglobin elicits a significant 72% protection in cattle. Analysis of immune responses to Fasciola during infection in ruminants suggests that chronic infection correlates with a type 2 helper T cell response, implying that type 1 helper T cell responses are down-regulated in fasciolosis. Recent results studying the resistance of Indonesian Thin Tail (ITT) sheep to F. gigantica have shown that this breed exhibits high innate (or rapidly acquired) resistance to infection and acquires a higher level of resistance after a primary challenge. Initial studies suggest that the resistance of ITT sheep to F. gigantica may be determined by a major gene. Merino sheep also acquire resistance to F. gigantica. In contrast, ITT and Merino sheep do not exhibit resistance to F. hepatica. These results suggest that there are fundamental differences between these two species of Fasciola in the biology of their interaction with the sheep immune system. In vitro studies on immune mechanisms of killing of juvenile fluke have shown that juvenile larvae of F. hepatica are susceptible to antibody dependent killing by activated rat macrophages in vitro which is mediated by nitric oxide. Future studies on the immune effector mechanisms expressed by resistant sheep which control infection by F. gigantica will lead to new knowledge which may allow the design of more effective vaccines for fasciolosis. PMID- 9394194 TI - The biological basis of malarial disease. AB - In this review we summarise the arguments that inflammatory cytokines, triggered by material released from the parasite at schizogony (malarial toxin), might induce the illness and pathology seen in malaria. These pro-inflammatory cytokines can generate inducible nitric oxide synthase and cause nitric oxide to be released, as can low concentrations of malarial toxin itself provided interferon-gamma, which has only low activity in the absence of malarial toxin, is present. We suggest here that recently described hypermetabolic functions of these mediators provide a much more plausible explanation for malarial hyperlactataemia and hypoglycaemia, the chief prognostic indicators in falciparum malaria, than does hypoxia secondary to mechanical blockage of vessels by sequestering parasites, which is the dominant current theory. We also review the arguments that rationalise, through these mediators, the reversibility of the coma of cerebral malaria. Although not yet tested at a cellular level, the proposal that nitric oxide generated in cerebral vascular walls contributes to this coma continues to gather indirect support. In addition, new evidence incriminating nitric oxide in the mechanism of tolerance to endotoxin rationalises the raised nitric oxide generation seen in malarial tolerance. PMID- 9394195 TI - Tumour necrosis factor and associated cytokines in the host's response to malaria. AB - Tumour Necrosis Factor (TNF) is produced at the initiation of malaria infections (pre-erythrocytic phase), as demonstrated by the release of bioactive TNF by peripheral blood mononuclear cells from individuals residing in endemic areas after stimulation with stage specific sporozoite antigens. During the erythrocytic phase, TNF production is greatly augmented by parasite antigens at the time of schizont rupture and merozoite release from infected erythrocytes. Some of the strongest inducers of TNF synthesis and release are malaria toxins, e.g. glycosylphosphatidylinositol moieties and malaria pigment. Because of TNF's well-known cytotoxic activity it was originally hypothesized that it alone was responsible for killing parasites directly or within host cells. Though earlier reports of the capability of serum containing TNF to kill plasmodia supported this idea, later experiments with recombinant TNF showed a lack of significant parasiticidal activity. Recent studies investigating related factors showed that they were involved with TNF in the control of infection. These factors included ther cytokines, such as interleukin (IL)-1, IL-6, IL-12, interferon-gamma (IFN gamma) as well as nitric oxide intermediates (NOI) and reactive oxygen intermediates (ROI). This positioned TNF as a key regulator of the immune response against the malaria parasite. However, it must be noted that TNF and its associated factors are also responsible for the fever, aches and pains of acute illness, as well as the hypoglycemia, shock, bleeding and reversible coma of severe malaria seen in approximately 1 percent of individuals with malaria. Therein lies the rub; factors important in the control of malaria also appear to have detrimental properties. Research presented in this review characterizes TNF and associated cytokines' importance in the immune response to malaria. PMID- 9394196 TI - An international practitioner data bank as a quality tool. AB - While international tort costs have not reached the level of those in the United States, international medical employers, like their American counterparts, must be able to ascertain updated objective information regarding the physicians they are going to employ, in order to protect their patients and organizations from the increasing risks in today's medical environment. The NPDB set up in the United States by the United States Congress provides a structure that can record and set standards for professional reviews. While the data bank established in the United States can still be considered a new entity, and the exact impact it has on quality, peer review and risk management is still being judged, it is the first step towards an organized, objective governing body. We recommend that an international committee convene to study the American model of a data bank and decide how and which parameters could be used for setting international standards and norms to be recorded in an international data bank. This data bank would be an important addition to the increasing array of tools available to ensure quality care. PMID- 9394197 TI - Data bank--"1984 and all that". PMID- 9394198 TI - International practitioner data bank as a quality tool: are we using behavior control in the right environment? PMID- 9394199 TI - Internationalizing the National Practitioner Data Bank. PMID- 9394200 TI - International Practitioner Data Bank: false premise, false promise. PMID- 9394201 TI - Improving health care quality. Is standardization the answer? PMID- 9394202 TI - The policy implications of using hospital and physician volumes as "indicators" of quality of care in a changing health care environment. AB - There is growing interest in the quality of health care and in using quality measures to direct patients to hospitals and providers offering high quality, low cost health care. The dilemma is that, while there is an increasing need for quality indicators as a result of a changing health care environment, this changing environment has important implications for the use of some of these measures. Since the 1970s, a growing body of research in the U.S. has addressed the empirical relationship between the number of patients with a specific diagnosis of surgical procedure and their outcomes after treatment in a particular hospital or by a particular physician ("volume-outcome" studies). In this paper, we examine the policy implications of using hospital and physician volume information as an "indicator" of quality in a rapidly changing health care environment with new players and new incentives. We begin by describing the evolution of the use of volumes within both regulatory and market-oriented contexts in the U.S. We then discuss policy considerations and cautions in using volumes, along with suggestions for future research. Our purpose is to point out potential problems and clarify confusions about the use of volumes, so that policymakers and practitioners can be sensitive to the potential minefields they are traversing. PMID- 9394203 TI - Treatment adequacy for HIV-related pneumocystis pneumonia: quality measures for inpatient care. AB - To develop and evaluate severity-adjusted indicators of treatment timeliness and adequacy for inpatient care of first episode of HIV-related pneumocystis pneumonia, a retrospective cohort study (n = 414) using medical record review was conducted in six California medical centers (1 January 1983-30 June 1987). Measures included patient baseline characteristics and complexity, process-of care indicators (delay in treatment initiation and proportion of adequate treatment delivered), and overall survival of hospitalization and survival without respiratory failure. Logistic regression models of severity were developed among optimally treated patients and cross-validated. Exposure to medication with pneumocystis activity within 30 days prior to admission was protective. After controlling for pre-admission medication and severity, the average proportion of adequate pneumocystis medication delivered during the first 7 and 30 days were significant predictors of outcome in all models. Delay in treatment initiation, while not a statistically significant predictor, was associated with baseline severity. Summary measures of treatment adequacy show promise as process-of-care indicators. PMID- 9394204 TI - Quality of STD care in Zambia. Impact of training in STD management. AB - STUDY OBJECTIVE: To assess quality of care of sexually transmitted diseases (STDs) and evaluate interactive training methods aimed at improving providers' performance. DESIGN AND SETTING: This comparative study, with a baseline, intervention, and evaluation phases was conducted at two urban health centers in Zambia. The personnel at one health center were trained in STD management using interactive training methods. The other health center acted as a control. SUBJECTS AND METHODS: Two-hundred patients with STD were interviewed and their interaction with health care providers observed before and after the training. Another 200 interviews and observations were conducted at the control health center. RESULTS: The proportion of patients being examined, given health education and informed about partner notification increased significantly after the intervention. The proportion of patients who had complaints about the health care did not decrease. Long waiting time and lack of time to discuss the disease were the main complaints. CONCLUSION: The training solved some, but not all, problems of poor case management. This indicates the need for a more process oriented approach for improving quality of care. PMID- 9394205 TI - Patient satisfaction with emergency house calls. AB - STUDY OBJECTIVES: To identify determinants of patient satisfaction with emergency house calls and to assess the properties of a satisfaction measurement questionnaire. DESIGN: Patient survey, combined with routinely collected information on the circumstances of the house call. SETTING: Emergency house calls provided by an independent emergency care organization (ECO) in Geneva, Switzerland. PARTICIPANTS: Consecutive sample of 389 patients (67% response rate). MAIN OUTCOME MEASURE: Patient satisfaction. PREDICTOR VARIABLES: patient age and sex, type of medical problem, time of visit, waiting time, duration of visit, perceived effectiveness of treatment. RESULTS: The satisfaction questionnaire was easy to administer. Factor analysis identified 3 separate dimensions of satisfaction, which pertained to the visit itself, to access and to general attitude toward the ECO. Validation tests were consistent with expectations. In multivariate analysis, older patient age and greater perceived treatment effectiveness predicted independently all satisfaction scales. Presence of a mixed physical and mental problem reduced satisfaction with the visit itself only, a delay between the phone call and the visit exceeding one hour reduced satisfaction with access and worsened the attitude toward the ECO. CONCLUSION: The instrument used to measure patient satisfaction with emergency house calls performed well. Overall levels of satisfaction were high. Perceived effectiveness of treatment was the strongest correlate of patient satisfaction. Monitoring of patient satisfaction in emergency settings may contribute to improvements of quality of care. PMID- 9394206 TI - Quality management savings: at Al-Hussein Hospital, Salt, Jordan. PMID- 9394207 TI - Quality Assurance Project: the second five years. PMID- 9394208 TI - Re: ISQua position paper. PMID- 9394209 TI - Re: ISQua position paper. PMID- 9394210 TI - "Every defect is a treasure". AB - After seeing multiple providers for various complaints, a patient becomes frustrated with the healthcare system. She is diagnosed with acromegaly secondary to pituitary tumor when she transfers to another healthcare system. PMID- 9394211 TI - Cold restraint-induced gastric lesions in individual- and group-stressed rats. AB - The aim of the present study was to 1) determine the intensity of cold restraint induced gastric lesions and core body temperature in single- and group-stressed rats, and establish a correlation between them; and 2) determine the influence of visual contact among animals during cold restraint on development of gastric stress ulcer. Therefore, adult male Wistar rats were put into individual or group restraint boxes (composed of two, three, six or nine single boxes) with or without possibility of visual contact and then exposed 2 hr to the cold (4 degrees C). Core body temperature was measured just before and after cold restraint using a digital rectal thermometer. The results showed that: 1) single stressed animals expressed significantly higher ulcer index than those stressed in group of three, six and nine rats; 2) there was no significant difference in degree of hypothermia among rats exposed to various group paradigms; and 3) there was no significant difference in ulcer index among animals stressed in conditions with or without visual contact. An absence of significant difference in ulcer index between single and paired stressed rats implies that three is the lowest number of animals per group at which an influence of group size on behavioral and adaptive mechanisms in rats exposed to cold restraint becomes manifest. PMID- 9394212 TI - Learning and memory in nucleus basalis magnocellularis-lesioned rats after transplantation of fetal frontal cortex. AB - The effect of fetal frontal cortex transplantation on behaviour performance was examined in adult male Wistar rats with lesions of the nucleus basalis magnocellularis (NBM). Compared to intact and sham-operated controls, the rats tested ten or twenty days after bilateral electrolytic lesions of NBM exhibited the significant learning and memory impairments (acquisition and performance of two-way active avoidance) whereas spontaneous motor activity was not significantly altered. The animals which received allotransplants of fetal frontal cortex (from 18-day gestational rat fetuses) into NBM, two ("early" transplantation-NBM-ET) or ten ("delayed" transplantation-NBM-DT) days after lesioning, respectively, manifested the complete amelioration of noticed impairments when tested ten days after transplantation procedure. Corresponding sham-transplants groups (NBM-SET and NBM-SDT) showed only slightly improvement of acquisition but not performance of two-way active avoidance. The ability of the transplants to restore learning and memory in the NBM lesioned rats suggests that graft of fetal frontal cortex can functionally influence neuronal activity of the lesioned host brain. PMID- 9394213 TI - Ultrastructural study of NADPH-d positive neurons in laminae I and II of the rat caudal spinal trigeminal nucleus. AB - The present study investigated the ultrastructure of neurons in the caudal spinal trigeminal nucleus. These neurons which are believed to function as interneurons in the transmission of orofacial nonreflexive nociceptive information, measured 20 microns x 11 microns, and were nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) positive. The reaction product, formazan, was localized in the nuclear envelope, mitochondria, rough endoplasmic reticulum, and multivesicular bodies of these neurons. It was also localized in the membrane of the smooth endoplasmic reticulum at the axon terminal. The neurons were contacted by both axosomatic and axodendritic synapses formed by both NADPH-d positive and NADPH-d negative axon terminals. Two types of NADPH-d positive axon terminals could be recognized. The first was a large terminal containing many stained mitochondria and unstained small round agranular vesicles mixed with some slightly flattened ones. It formed asymmetrical axodendritic synapse. The second type of axon terminals contained pleomorphic synaptic vesicles and formed asymmetrical synapses upon both dendrites and soma. The sources of NADPH-d positive axon terminals were discussed. Most of the unstained axon terminals forming axosomatic and axodendritic synapses with stained cell bodies and dendrites contained flattened vesicles. In addition to the above, complicated synaptic configurations showing NADPH-d positive axoaxonic synapses in relation to NADPH-d negative dendritic spines were also seen in which a NADPH-d negative dendritic spine was completely contacted by a NADPH-d positive bouton which was in turn contacted by another NADPH-d positive bouton. PMID- 9394214 TI - Impaired temporal discrimination in Parkinson's disease: temporal processing of brief durations as an indicator of degeneration of dopaminergic neurons in the basal ganglia. AB - Recent findings suggest that temporal processing of brief durations is a function of dopaminergic neurotransmission in the basal ganglia. Furthermore, there is preliminary evidence of abnormal timing functions in patients suffering from idiopathic Parkinson's disease (PD). In the present study, temporal discrimination of intervals in the range of milliseconds was investigated in 20 PD patients and 20 healthy controls matched for sex and age. Temporal discrimination was significantly impaired in PD patients as compared to healthy controls. For PD patients, additional correlational analyses did not yield any significant relationship between performance on temporal processing and degree of motor impairment or illness duration. However, impairment in temporal processing was associated with dosage of L-dopa substitution and self-rated feelings of depression. The overall pattern of results suggests that deficits in temporal information processing observed in PD patients may represent a trait marker of vulnerability to decreasing levels of dopaminergic activity in the basal ganglia rather than a state-dependent indicator of the acute clinical symptomatology. PMID- 9394215 TI - Reversal of cognitive impairment in an elderly parkinsonian patient by transcranial application of picotesla electromagnetic fields. AB - A 74 year old retired building inspector with a 15 year history of Parkinson's disease (PD) presented with severe resting tremor in the right hand, generalized bradykinesia, difficulties with the initiation of gait with freezing, mental depression and generalized cognitive impairment despite being fully medicated. Testing of constructional abilities employing various drawing tasks demonstrated drawing impairment compatible with severe left hemispheric dysfunction. After receiving two successive transcranial applications, each of 20 minutes duration, with AC pulsed electromagnetic fields (EMFs) of 7.5 picotesla flux density and frequencies of 5Hz and 7Hz respectively, his tremor remitted and there was dramatic improvement in his drawing performance. Additional striking improvements in his drawing performance occurred over the following two days after he continued to receive daily treatments with EMFs. The patient's drawings were subjected to a Reliability Test in which 10 raters reported 100% correct assessment of pre- and post drawings with all possible comparisons (mean 2 = 5.0; p < .05). This case demonstrates in PD rapid reversal of drawing impairment related to left hemispheric dysfunction by brief transcranial applications of AC pulsed picotesla flux density EMFs and suggests that cognitive deficits associated with Parkinsonism, which usually are progressive and unaffected by dopamine replacement therapy, may be partly reversed by administration of these EMFs. Treatment with picotesla EMFs reflects a "cutting edge" approach to the management of cognitive impairment in Parkinsonism. PMID- 9394216 TI - Occurrences of electroencephalographic (EEG) patterns that resemble epileptiform discharges in background EEG in epileptic patients. AB - The relationships of background EEG to epileptiform discharge development were studied in 9 epileptic patients having generalized spike and wave complexes (SWCs) with a maximum at the frontal location on either side. The instantaneous power spectrum of EEGs at F3, F4, C3, C4, P3, P4, O1, O2, T3 and T4 was estimated with wavelet transform. Subsequently, the similarity of power spectra between SWCs and background EEG was determined by use of Kullback-Leibler information and artificial neural network in each patient. Both similarity measures revealed that EEG patterns similar to SWCs occurred in the background activity just before SWCs at frontal locations. These findings suggested that these SWC-like EEG events occurred as poorly developed epileptiform discharges buried in the background activity. PMID- 9394217 TI - The Stroop's test evokes a negative brain potential, the N400. AB - The Event Related Potential (ERP) of 8 french right handed subjects were recorded with 5 active electrodes located in frontal (Fz), central (Cz), occipital (Oz) and right/left parietal (RH, LH) sites while they were performing a modified version of the test of Stroop. They had either to read the names of basic colors (yellow, green, blue, red) written in the same colors (red written in red: concordant stimuli) or in a different color (red written in blue: discordant stimuli) or to name mentally the color in which was written the name of a color, both colors being concordant or discordant. The ERPs for reading were similar for concordant and discordant stimuli and showed no sign of a N400 wave, this was also the case for the mental naming of a color associated to the written name of the same color. A N400 wave with a Cz location was evident for the mental naming of a color when it was associated to the written name of another color. In this last case, the automatic reading of the name of a color would correspond to a priming which interferes with the access to the target word: the name of another color that the subject is required to evoke mentally. PMID- 9394218 TI - Brain-stem auditory evoked potential monitoring. The increase of the stimulus artifact in the development of brain death: a biological phenomenon? AB - Brain-stem auditory evoked potentials (BAEPs) were recorded in 12 dead subjects (mean age, 72.6 +/- 14.8 years), 30.6 +/- 19.5 hours (range 9-70) after abolished systemic circulation. Death was due to cardiac failure (n = 10), intracerebral hemorrhage (n = 1) and larynx cancer (n = 1). The presence and amplitude of the stimulus artifact were evaluated. The mean (+/- SD) amplitudes of the stimulus artifact was 0.03 +/- 0.02 microV on the left side and 0.01 +/- 0.02 microV on the right side. These findings in accordance with previous studies on comatose patients and brain dead subjects confirm that the increase of the stimulus artifact in the development of brain death, in spite of stimulation with alternating polarity, seems to reflect a biological phenomenon which is not found in dead subjects after complete cessation of systemic circulation. PMID- 9394219 TI - Electrophysiological analysis of expectancy: P3 in informed guessing. AB - 18 healthy subjects had to guess, which of two nonequiprobable events would occur next. Each trial was preceded by a cue which increased the probability of the corresponding event as compared to its global probability. Event-related potentials (ERPs) were recorded and then classified according to global probability of events, their relation to the preceeding cue (valid versus invalid cues) and to subject's prediction (predicted versus nonpredicted). Two late positive waves (P350 and P550) with parietal maxima were distinguished. Both waves had larger amplitudes in response to improbable events than to highly probably events. Similarly, both had larger amplitudes following invalid cues than following valid cues, and this difference was larger in those subjects who tended to follow the cue than in those who tended to reject it. No difference in terms of ERP component amplitudes was found between predicted and unpredicted events; however, the latency of the P350 peak was longer following unpredicted events. Taken together with data of the literature, the present results indicate that ERP allow us to distinguish between two meanings of the word "expectancy": (1) the rule-related expectancy as cognitive estimation of the likelihood or "representativeness" of an event (based on grasping event contingencies), and (2) the goal-related expectancy manifested in the subject's overt behavior. Only the former "expectancy" affects the amplitude of the late positive wave ("P3"), while the latter does not. PMID- 9394220 TI - Reliability of dipole localization for the movement-evoked field component MEF I. AB - The movement-evoked field I (MEF I) component is the largest and most stable neuromagnetic component accompanying self-paced movements. In order to use MEG for studying dynamic changes in the cortical organization of movements, data about the reliability and variability of these neuromagnetic components for individual subjects must be established during different sessions. For this aim, three male subjects were requested to perform self-paced flexions of their index finger and thumb in repeated sessions while the MEG was recorded by a 31 channel system. The MEF I was identified for each session and a single equivalent dipole was calculated for this component. The dipole localizations of the various sessions were compared. The standard deviation of the localization for all persons and all values amounts to 4.0-5.2 mm for the three spatial dimensions. Our data suggest that the spatial distance between two single focal sources fitted to the MEF I must be greater than 14 mm to be interpreted as distinct. However, the neuromagnetic field structure and the resulting dipole localization of the MEF I component are quite stable and could be used for the evaluation of cortical plasticity. PMID- 9394221 TI - Modulation of microglial form and immune function by factors released from goldfish optic nerves. AB - Activation of microglia is associated with neural damage and may aid repair of the CNS. To begin to investigate their role, microglia purified from mouse brain were grown in media conditioned (CM) by goldfish optic nerve (GFON), optic tectum (GFOT), vagal lobe, telencephalon and cerebellum, and medium conditioned by rat optic nerves (RON). Microglia maintained in GFON- or GFOT-CM assumed an ameboid morphology, whereas microglia grown in media conditioned by the other neural tissues produced long, crenellated processes that resembled the ramified microglial form. Microglia maintained in all types of CM functioned as antigen presenting cells in a MHC-restricted manner when tested on conalbumin-specific Thelper (Th) cells, except for microglia maintained in GFON- and GFOT-CM. These studies suggest that GFON, in contrast to RON, produces a substance(s) that affects microglial morphology and immune reactivity, and may promote the vigorous regeneration seen in GFON after damage. PMID- 9394222 TI - Facial asymmetry in right- and left-handed men and women. AB - A posteroanterior cephalometric radiographic study was performed on the right- and left-handed men and women with normal occlusion. A posteroanterior cephalometric radiography was conducted in these subjects. Method of triangulation was used to measure various face areas. The surface areas of these triangles were compared with their equivalents on the contralateral side. Sex and its interactions with handedness and side were significant factors influencing facial areas. Areas on the left were found to be significantly larger than those on the right in right-handers. Left-handers were inconsistent in facial asymmetry, but they tended to have larger facial areas on the right than the left. Sex was especially significant for left-handers. It was suggested that an asymmetric development in some brain regions may be responsible for the development of asymmetric facial regions. PMID- 9394223 TI - Case report: subependymal diffuse heterotopia. Clinical, EEG, and neuroimaging findings. AB - We report the case of a 41 year-old woman with a slight mental retardation and epilepsy. MRI showed a diffuse subependymal heterotopia. The cognitive level supports the view that the ectopic cells are probably not important for the normal cortical functions but that they are likely able to maintain some of the electric properties of normal neurons, even if with altered discharge modalities. The genetic etiology of subependymal neuronal migration disorders is discussed. This is the second reported case of diffuse subependymal heterotopia in a female patient. PMID- 9394224 TI - Effects of intrathecal monoamine antagonists on the nociceptive c-Fos expression in a lesioned rat spinal cord. AB - Effects of intrathecal (i.t.) administration of monoamine antagonists on formalin induced neuronal c-Fos expression in two sides of the lumbar dorsal horn were observed in rats with unilateral transection of the dorsolateral funiculus at T11 12 level. The results showed that: 1) pretreated with i.t. normal saline (control) and then an equal volume of formalin was injected into the two hindpaws, the number of Fos-like immunoreactive neurons were 44% lower on the side of lumbar dorsal horn with intact dorsolateral funiculus (57 +/- 3.1 vs. 103 +/- 3.8). 2) Pretreatment with i.t. phentolamine (a non-selective alpha adrenoceptor antagonist) caused an increase of Fos-like immunoreactive neurons on the intact side so showing only a reduction rate of 23% to the lesioned side (p < .01); 3) pretreatment with i.t. cyproheptadine (a 5-HT-receptor antagonist) caused a similar reduction rate of 21% (p < .01) of Fos-like immunoreactive neurons on the intact side; and 4) combined i.t. pretreatment with phentolamine and cyproheptadine caused a reduction of Fos-like immunoreactive neurons of only 4% on the intact side, namely, the differences in the number of Fos-like immunoreactive neurons on two sides of the lumbar spinal cord owing to the unilateral dorsolateral funiculus lesion were nearly abolished by i.t. coinjection of phentolamine and cyproheptadine. The results indicate that 1) peripheral noxious inputs can provoke a spinally-descending inhibitory effect on the spinal nociceptive transmission via the dorsolateral funiculus and 2) the descending fibers in dorsolateral funiculus exert their action mainly through the release of either norepinephrine or 5-HT at the spinal level. PMID- 9394225 TI - Open field behavior in nucleus basalis magnocellularis-lesioned rats treated with physostigmine and verapamil. AB - The present study was done to investigate and compare the effect of acetylcholinesterase inhibitor, physostigmine (0.030, 0.045, 0.060 and 0.075 mg/kg sc) and Ca-antagonist, verapamil (1.0, 2.5, 5.0 and 10.0 mg/kg sc) on open field behavior in male Wistar rats with bilateral electrolytic lesions of nucleus basalis magnocellularis (NBM). NBM-lesions produced a significant increase and decrease of ambulation and number of inner squares entered, and defecation, respectively, with no influence on grooming in rats exposed to novel environment. Physostigmine and verapamil in all tested doses, given 30 min before the test did not affect the open field behavior in control animals. In contrast to that, physostigmine (0.045, 0.060 and 0.075 mg/kg) and verapamil (2.5 and 5.0 mg/kg) significantly reduced ambulation and number of inner squares entered in NBM lesioned rats. Also, physostigmine in a dose of 0.060 mg/kg significantly decreased defecation and in doses of 0.060 and 0.075 mg/kg the grooming, as well. On the other hand, verapamil only in a dose of 2.5 mg/kg significantly increased defecation. It could be concluded that lesions of NBM in rats induced disturbances in the open field behavior, which might be successfully ameliorate by physostigmine and verapamil treatment. PMID- 9394226 TI - Treatment with AC pulsed electromagnetic fields improves the response to levodopa in Parkinson's disease. AB - A 52 year old fully medicated Parkinsonian patient with severe disability (stage 4 on the Hoehn & Yahr disability scale) became asymptomatic 10 weeks after he received twice weekly transcranial treatments with AC pulsed electromagnetic fields (EMFs) of picotesla flux density. Prior to treatment with EMFs, his medication (Sinemet CR) was about 50% effective and he experienced end-of-dose deterioration and diurnal-related decline in the drug's efficacy. For instance, while his morning medication was 90% effective, his afternoon medication was only 50% effective and his evening dose was only 30% effective. Ten weeks after introduction of treatment with EMFs, there was 40% improvement in his response to standard Sinemet medication with minimal change in its efficacy during the course of the day or evening. These findings demonstrate that intermittent, AC pulsed applications of picotesla flux density EMFs improve Parkinsonian symptoms in part by enhancing the patient's response to levodopa. This effect may be related to an increase in the capacity of striatal DA neurons to synthesize, store and release DA derived from exogenously supplied levodopa as well as to increased serotonin (5-HT) transmission which has been shown to enhance the response of PD patients to levodopa. Since decline in the response to levodopa is a phenomenon associated with progression of the disease, this case suggests that intermittent applications of AC pulsed EMFs of picotesla flux density reverse the course of chronic progressive PD. PMID- 9394227 TI - Drug induced double peak waveforms in the N20-component of somatosensory evoked potentials. AB - The double peak waveform of the N20 component of the early somatosensory evoked potentials is a rare finding. In the present paper we investigated 9 patients (mean age +/- SD: 31.3 +/- 11.3 years; range 18-52 years) with the phenomenon of the two subcomponents of the primary cortical complex, after stimulation of the median nerve in a retrospective analysis. The results of the heterogenous patient group showed that the generation of the second subcomponent is not pathognomonic for patients with severe head injury, that it could be reversible and that pharmacological induced effects are presumably responsible for this phenomenon. PMID- 9394228 TI - Involvement of brain dopaminergic structures in neuroimmunomodulation. AB - Bilateral electrolytic destruction of the brain areas containing dopamine (DA) cell bodies (nuclei A9 and A10) as well as terminal regions of the nigrostriatal and mesolimbic DAergic systems (nuclei caudatus and accumbens) resulted in a considerable decrease in the intensity of the immune response in rats immunized with sheep red blood cells (SRBC). Administration of SRBC (5 x 10(8) i.p.) to rats produced a marked rise in activity of central DAergic system at early stage of the immune response formation. The most pronounced elevation in the concentration of DA and its metabolites, measured by the method of high performance liquid chromatography with electrochemical detection, was observed in the terminal regions of the nigrostriatal and mesolimbic DAergic systems (nuclei caudatus and accumbens), hypothalamus, hippocampus, amygdala within 20 min following antigen inoculation. By 60 min after immunization DA metabolism has been retained at a high level in all brain regions examined. The concentration of DA returned to control level in the amygdala and hypothalamus 24 hours after antigen administration and had a tendency to reach control values in the rest of the structures. The present results indicate that nigrostriatal and mesolimbic DAergic systems and DAergic structures of the hypothalamus are involved in the mechanisms of neuroimmunomodulation. PMID- 9394229 TI - Unilateral brain lesions and performance on Russell's version of the Wechsler Memory Scale in an African American population. AB - Studies of patients with unilateral lesions report hemisphere-specific and locus specific impairments on Russell's (1975) Revision of the Wechsler Memory Scale (RWMS). In the current investigation "race-homogeneous" and "race-comparative" paradigms provide the context in which the generalizability of RWMS findings are examined in a population of African Americans with unilateral lesions. The performances of brain-damaged patients were impaired relative to normal controls on five of the six RWMS measures. However, patients with left and right hemisphere damage in our sample did not differ systematically on RWMS subtests. Likewise, among patients with lesions confined to one of the quadrants in the brain, there were no quadrant group differences in performance on RWMS subtests. But, right posteriors were impaired relative to controls on immediate and delayed VR subtests. The relative merits of the race-comparative and race-homogeneous paradigms are considered in the context of these findings. PMID- 9394230 TI - Attentional systems and the allocation of cerebral resources in reading and grammatical tasks. AB - To evaluate the possible role of attentional centers as modulators of neural networks that mediate visual tasks involving reading and grammatical manipulations of verbs, we measured cerebral blood flow (CBF) using positron emission tomography (PET), and reaction times as subjects read verbs, "nonce verbs" such as jelt or brep, and formed past tenses of regular, irregular and nonce verbs after viewing their stems. Statistical parametric maps (SPMs) showed significant activation of the pulvinar in the read verb irregular, and generate nonce past tense tasks, compared to rest. This was confirmed by a post hoc ANOVA of CBF values from a discrete locus in the pulvinar (p = .0000417). Functional links between the pulvinar and other brain regions were shown by high correlations of CBF in the pulvinar with CBF in brain regions known to have anatomical connections to the pulvinar, particularly those mediating vision. There was also a significant relationship between task-specific reaction times and rest minus task CBF differences in a multiple regression analysis that included CBF values from the pulvinar, superior colliculus plus reticular formation, and the anterior cingulate, known attentional centers (p = .021, r2 = 0.99). Regression analyses relating reaction time to the amount of brain activated (pixels in the SPMs) and the degree of activation of the pixels (mean Z score) yielded p values of .078 and .074, respectively. Our data provide direct experimental evidence to support the hypothesis that attentional centers are activated in proportion to the complexity of visually mediated language tasks and that the centers that mediate attention modulate the activity of task-specific neural networks. PMID- 9394231 TI - Evidence of altered cerebral blood-flow relationships in acute phobia. AB - Functional cerebral guiding and integrating systems may be revealed by analyzing the covariation of regional cerebral blood flow (rCBF). Positron emission tomography (PET) was used to measure absolute rCBF in 14 volunteers with specific phobia and 6 nonphobic controls, when exposed to videos containing phobia relevant and neutral scenes. A fear reaction and increased covariation between absolute rCBFs was observed during phobia-relevant as compared to neutral stimulation in phobics only. In controls fear was not elicited and rCBF covariation was not influenced by stimulus condition, being similar to the pattern observed in phobics during neutral stimulation. We suggest the rCBF correlative pattern during phobic fear to reflect fear-related activation of distinct neuronal pathways that involves the amygdala, the thalamus, and the striatum. We theorize that these pathways are activated also by uncontrolled emotions in diverse conditions, like posttraumatic stress disorder, panic disorder, and schizophrenia. PMID- 9394232 TI - Clinical case report: memory functions after anterior communicating artery aneurysm rupture. AB - The case of a patient suffering from rupture of an aneurysm of the anterior communicating artery is reported. The hematoma was located in the basal forebrain, one of the anatomical regions associated with mnestic functions. Memory and other cognitive functions were studied. The patient performed well on all non-mnestic tests, but showed significant memory impairment. Anterograde memory was deficient not only in recall, but also in recognition. Performance on remote memory tests showed some differentiation. The patient remembered autobiographical material and famous faces correctly, but presented deficits on a Famous Events Test. This dissociation represents a difference in emotional content or familiarity of the material; the performance in tests using material with a higher emotional content was unremarkable. Lesions within the basal forebrain seem to deteriorate anterograde and remote memory, but performance may be inconspicuous if test material of high emotional content or familiarity is used. PMID- 9394233 TI - Encoding into working memory of spatial location, color, and shape: electrophysiological investigations. AB - Event-related potentials (ERP) were recorded while subjects memorized either the location, the color or the shape of stimuli which could be located in 1 of 4 positions relative to a central fixation point (top, bottom, left or right), be of 1 of 4 positions relative to a central fixation point (top, bottom, left or right), be of 1 of 4 colors (white, green, red or blue), and present 1 of 4 shapes (triangle, cross, circle or square). These ERP were compared to ERP recorded while subjects looked at the same stimuli but performed other control, nonmemory tasks. Only ERP corresponding to the memorization of spatial location showed a differential pattern which could be specifically attributed to memory encoding processes. This reveals an important difference in ERP modulation between a working memory subsystem for spatial location and other subsystem (or subsystems) for color or shape, which would provide evidence supporting the existence of different working memory subsystems for visual information in the brain. PMID- 9394234 TI - Visual organization test performance in an African American population with acute unilateral cerebral lesions. AB - Controversy abounds as to whether the Hooper Visual Organization Test (VOT) is a measure of hemisphere-specific, region-specific, or non-specific brain damage. The present study examines this issue in a group of African Americans with acute unilateral brain damage and non-brain-injured controls. Consistent with the idea that the VOT is a measure of "organic" cerebral pathology, non-brain damaged controls earned significantly higher VOT scores than brain-damaged patients. While other studies have noted that the VOT is primarily sensitive to damage in the right parietal region of the brain, the present study shows that VOT performance is especially vulnerable to acute lesions in the right anterior quadrant of the brain. This latter finding supports the idea that VOT performance is differentially sensitive to regional cerebral pathology, but challenges the region specific claim of poorer VOT performance among patients with right posterior cerebral damage. PMID- 9394235 TI - [Clinical evaluation of faropenem against infections in pediatric fields]. AB - The recent increases in the prevalence of penicillin-resistant Streptococcus pneumoniae becomes a point at issue clinically. We carried out a clinical study in 40 cases in the pediatrics department, as faropenem (FRPM) was proved to have an excellent antimicrobial activity against penicillin-resistant Streptococcus pneumoniae. The study was planned to investigate in detail the movement of stools that had been a problem in a clinical development studies out before. In this study, an observation of the daily movement of stools was one of the principal evaluation items, hence the patients were divided into two groups. One group (S group) were administered FRPM only, the other group (E-group) were administered FRPM in combination with a medicine for intestinal disorders (Enteronon-R). An observed frequencies of any loose bowel movements were 94.7% in S-group, and 63.2% in E-group, hence the study suggested that the combination drug was effective. The patients observed higher frequencies of development of the movement of stools, all of them were recovered from in the course of administration or within 4 days after administration, however whether or not being treated symptomatic therapy. Clinical efficacy rates of FRPM on mainly respiratory infections were 94.6%. In this study, 4 strains (patients) of penicillin-resistant Streptococcus pneumoniae were isolated. Against penicillin resistant Streptococcus pneumoniae, FRPM demonstrated more potent antibacterial activity than the oral penicillins and cephems tested here except cefditoren. Clinical efficacies was deemed effective in all of the 4 cases, and bacteriologically, 3 organisms were eradicated. As for side effects including diarrhea and loose stool, no serious side effects were observed. Based on the above results, FRPM is effective against most infections in the pediatric field which Streptococcus pneumoniae are isolated at high frequencies highly, and is considered to cases in be useful an attention will have to be paid to stool movement, however. PMID- 9394236 TI - [Clinical and bacteriological effects of cefetamet pivoxil against community acquired respiratory tract infections. Part II]. AB - We investigated clinical and bacteriological effects of cefetamet pivoxil (CEMT PI) in community-acquired respiratory tract infections and obtained the following findings. That method was approximately equal to that of investigation in 1994. 1. Of the 431 respiratory tract infection cases that were treated with CEMT-PI according to a same protocol at a total of 41 institutions in Tokyo, Kanagawa ken, Saitama-ken and Chiba-ken from January to the beginning of March 1996. Outpatients accounted for 98.1% of the subjects. Regarding genders to patients, slightly more females (52.6%) than males were included. Diagnoses given to these patients included pharyngo-laryngitis (53.5%), tonsillitis (20.4%) and acute bronchitis (19.1%). 2. We investigated clinical efficacy rates (the ratio of those excellent + good) classified by diseases. The improvement rates of pharyngo laryngitis, tonsillitis and acute bronchitis were more than 85.0%. Other cases were small in number. That of chronic bronchitis-acute increasing change for the worse was 66.7%, pneumonia was 50.0% and bronchiectasis infection was 16.7%. It was not studied that clinical efficacy rates among those who were treated with 1 CEMT-PI tablet twice and among those who were given 2 tablets twice were significant level. 3. For the bacteriological study, a written material describing the method of collecting specimens, storage and transport in detail was distributed to the above mentioned institutions. The isolation and identification of suspected causative bacteria, determination of minimum inhibitory concentrations (MICs) and investigation of beta-lactamase production were conducted all together at section of studies, Tokyo Clinical Research Center. Suspected causative bacteria were detected from 274 (63.6%) cases. They included 88 strains of Haemophilus influenzae, 47 strains of Streptococcus pneumoniae, 42 strains of Streptococcus pyogenes, 20 strains of Moraxella subgenus Branhamella catarrhalis and 17 strains of Klebsiella pneumoniae subsp. pneumoniae. Suspected causative bacteria classified by diseases were S. pyogenes (tonsillitis), S. pneumoniae (acute bronchitis and secondary infection of chronic respiratory infection) and H. influenzae (pharyngo-laryngitis), and the detection frequency of those was high. The clinical efficacies (the ratio of improvement) classified by suspected causative bacteria were 84.4% against organism that was indicating CEMT and were 69.2% against organism that was not indicating CEMT. PMID- 9394237 TI - [Recent trends in incidence of respiratory tract pathogens and antimicrobial susceptibilities of Haemophilus influenzae, Streptococcus pneumoniae and Moraxella catarrhalis isolated in 1994 and 1995]. AB - The incidence of pathogenic bacteria in respiratory tract infections in 1994 and 1995 was investigated using quantitative cultures of sputa from patients with the infections in our department. Haemophilus influenzae, Streptococcus pneumoniae and Moraxella catarrhalis were isolated at high rates (70.5% in 1994 and 73.8% in 1995) from the specimens of out-patients, and the incident rates were similar to the past data. The antimicrobial susceptibilities of these three pathogens were examined with the agar dilution method. The incidence of penicillin (Pc) resistant S. pneumoniae against which MIC of Pc-G was higher than 0.125 microgram/ml was markedly increased from 24% in 1994 to 34.9% in 1995. Most of the Pc resistant isolates were also resistant to other antibiotics including erythromycin, minocycline and tosufloxacin. Serotype of strains against which MIC of Pc-G was higher than 1.0 microgram/ml was 19. The ratios of beta-lactamase producing strains among H. influenzae isolated in 1994 and 1995 were 20 and 15.8%, respectively, which were slightly higher than those in the past. One quinolone resistant strain was isolated in this study. Although the ratio of beta lactamase-producing strains among M. catarrhalis was as high (96.7%) as in the past, no increased resistance against the drugs examined was observed. PMID- 9394238 TI - [Antimicrobial activities of clarithromycin against recent obtained clinical isolates]. AB - In order to evaluate antimicrobial activities of clarithromycin (CAM), minimum inhibitory concentrations (MICs) of CAM and control drugs were determined against clinical isolates that were obtained from outpatients in 1994 and 1996. The results are summarized as follows; 1. It was not showed that CAM-resistant strains were increasing among Staphylococcus spp., beta-streptococci, Moraxella subgenus Branhamella catarrhalis, Haemophilus influenzae, Bordetella pertussis, Campylobacter jejuni subsp. jejuni, Chlamydia trachomatis and Mycoplasma pneumoniae. It appeared that resistances to CAM and macrolides (MLs) were increasing among Streptococcus pneumoniae and Peptostreptococcus spp. 2. The drug susceptibility patterns to MLs were similar and detection frequencies of induced resistant strains that were resistant to only 14-membered ring MLs including CAM and constitutive resistant strains that were resistant to 14 and 16-membered ring MLs were high among Streptococcus pneumoniae and Peptostreptococcus spp. It appears that MLs-resistance systems are linked to each other, and that this was a cause of increasing MLs-resistance among these bacterial species. 3. Notwithstanding of antibiotic resistance problems, CAM is still useful since it maintains strong antimicrobial activities against M. (B.) catarrhalis, B. pertussis, C. jejuni subsp. jejuni, C. trachomatis and M. pneumoniae, and it controls arginate producing abilities of mucoide strains of Pseudomonas aeruginosa. PMID- 9394239 TI - [Evaluation of reduced vancomycin susceptibility of MRSA strain Mu50 with various conditions of antibiotic susceptibility tests]. AB - We evaluated a clinical strain of methicillin-resistant Staphylococcus aureus (MRSA), Mu50, for vancomycin susceptibility. Mu50 was isolated from a patient with infection of a surgical incision site which resisted vancomycin therapy. Mu50 showed a decrease in susceptibility to vancomycin, but this strain did not carry vanA or vanB or vanC genes as judged from PCR amplification. MICs of vancomycin against Mu50 and vancomycin-susceptible S. aureus FDA209P, S. aureus ATCC-29213, and MRSA H-1 were 8, 1, 1, and 1 microgram/ml, respectively by agar dilution and macro-broth dilution methods according to NCCLS. MIC values with agar dilution method using MHA + 20% horse serum, HIA, and BHIA agreed with the MIC values with micro- and macro-broth dilution method. Population analysis revealed that vancomycin concentration required for inhibition of ca. 10(7) cells of Mu50, S. aureus FDA209P, S. aureus ATCC29213, and MRSA H-1 were 36, 2, 2, and 2 micrograms/ml, respectively. These results showed that the activity of vancomycin against Mu50 was at least 8-fold decreased compared to that against S. aureus FDA209P, S. aureus ATCC29213, and MRSA H-1. PMID- 9394240 TI - [The in vitro antifungal activities of fluconazole against pathogenic yeasts recently isolated from clinical specimens]. AB - The emergence of Candida albicans resistance to azole antifungal agents have been reported in the U. S. and Europe. We examined the in vitro antifungal activities of fluconazole against clinical isolates collected by seven investigators in three years to examine if a tendency existed toward the development of azole resistance among fungal isolates in Japan. The following results were obtained: 1. Sensitivities to fluconazole (FLCZ) were determined for yeast-like fungi, including 113 strains isolated in 1993, 149 strains isolated in 1994 and 205 strains isolated in 1995. No significant differences in sensitivities in the three years were detected. 2. Minimum inhibitory concentrations of FLCZ were 0.1 0.78 microgram/ml for C. albicans and 3.13-25 micrograms/ml for C. glabrata. Strains with 25 micrograms/ml of FLCZ's MIC were detected; two strains of C. krusei and one strain each of C. krusei, Trichospron beigelii and Hansenula anomala. No strains with higher than 50 micrograms/ml MIC of FLCZ were detected. 3. In vitro activities of FLCZ were compared between clinical strains isolated between 1993 and 1995 and clinical strains isolated before the marketing of FLCZ (up to December 1987) or clinical yeasts isolated between 1991 and 1992. No significant differences were observed, suggesting that no tendency existed toward azole resistance among fungal strains examined. PMID- 9394241 TI - Usage of T cell receptor (TCR) V beta gene in ulcerative colitis. AB - Ulcerative colitis (UC) is an inflammatory disease of unknown etiology. In this study, the expression of TCRV beta gene in UC patients was examined. The present study included 11 consecutive Japanese patients with UC. The control group comprised 10 healthy people. The usage of T Cell Receptor (TCR) V beta chain gene segments in peripheral blood was examined using RT-PCR. In addition, HLA-DR DNA typing was performed for the 11 patients with UC. The average of the positive expression rates in the UC group was 0.617 +/- 0.126, which was significantly higher than that of the control group (0.829 +/- 0.080) (p < 0.01). In the UC group, the usage of V beta chain gene in the active phase was almost same as that in the inactive phase. No specific distribution of V beta repertoire was observed in the UC group. In UC patients, no distinct correlation was found between any certain HLA-DR type and TCRV B gene usage. UC patients had a significantly lower rate of the expression of TCR V beta genes compared to normal controls. There is a possibility that the restricted usage of TCR V beta repertoire is possibly related to the existence of oligoclonal TCR repertoire in UC patients. PMID- 9394242 TI - Adjuvant activity of diesel exhaust particulates (DEP) in production of anti-IgE and anti-IgG1 antibodies to mite allergen in mice. AB - The present study indicates that diesel exhaust particulates (DEP) and pyrene contained in DEP have an adjuvant activity on IgE and IgG1 antibody productions in mice immunized intranasally with a mite allergen. The effect of pyrene on IgE and IgG1 antibody productions in mice was investigated to clarify the relation between mite allergy and adjuvancy of the chemical compounds in DEP. Der f II, one of the major allergens of house dust mite (Dermatophagoides farinae), was used as a mite allergen. Mice were grouped, and immunized with 5 micrograms of Der f II alone, 5 micrograms of Der f II plus 200 micrograms of pyrene and 5 micrograms of Der f II plus 100 micrograms of DEP intranasally seven times at two week intervals. The separate groups of mice were also immunized with 10 micrograms of Der f II plus the same dose of adjuvants in the same way. The IgE antibody responses to Der f II in mice immunized with Der f II plus pyrene or Der f II plus DEP were markedly enhanced compared with those immunized Der f II alone. The anti-Der f II IgE antibody production increased with increasing the dose of Der f II from 5 micrograms to 10 micrograms in mice immunized with Der f II plus the same dose of adjuvants. The IgG1 antibody responses to Der f II in mice immunized with 10 micrograms of Der f II plus 200 micrograms of pyrene or 10 micrograms of Der f II plus 100 micrograms of DEP were extremely higher than those immunized with 10 micrograms of Der f II alone. In addition, when the peritoneal macrophages obtained from normal mice were incubated with pyrene or DEP in vitro, an enhanced interleukin-1 alpha production of the macrophages was observed. When the spleen lymphocytes obtained from the mice immunized with 10 micrograms of Der f II plus 100 micrograms of DEP or 10 micrograms Der f II plus 200 micrograms of pyrene were stimulated with 10 micrograms of Der f II in vitro, an enhanced IL-4 production of the lymphocytes was also observed compared with those immunized with Der f II alone. These results suggest that the adjuvancy of DEP and pyrene on the production of IgE and IgG1 antibodies to Der f II may be one of the factors responsible for an incidence of asthma caused by house dust mite. PMID- 9394243 TI - Characterization of reactivity of monoclonal autoantibodies with renal antigens in experimental lupus nephritis. AB - Mice with chronic graft-versus-host disease (GvHD), induced by injection of DBA/2 lymphocytes into (C57BL10*DBA/2)F1 hybrids, develop a lupus-like syndrome with immune complex glomerulonephritis. Circulating autoantibodies are reactive with various self-antigens, including DNA, renal tubular epithelium (RTE), and laminin 1. To elucidate the reactivity of autoantibodies with renal antigens in experimental lupus nephritis further, the reactivity of the autoantibodies was studied in more detail by generating hybridomas from GvHD spleen cells. Hybridomas were selected for reactivity with RTE and laminin-1 coated on nitrocellulose sheets. Four stable clones were obtained (GV1-GV4). Monoclonal antibody (mAb) GV1 showed no reactivity on kidney sections, while GV2 stained the brush border of proximal tubular epithelial cells. Both GV1 and GV2 reacted only with RTE in ELISA. GV3 showed a nuclear staining pattern, while GV4 stained matrix structures on F1 kidney sections. GV3 and GV4 both reacted with RTE, laminin-1, ssDNA, and dsDNA in ELISA. Growth of hybridomas in mice, but not passive transfer of the mAbs, led to glomerular Ig binding for mAbs GV3 and GV4 without development of proteinuria. Our results show that in addition to anti nuclear autoantibodies cross-reactive with renal antigens, autoantibodies reactive with renal antigens and not with DNA are generated during chronic GvHD. Based on these results, combined with those of earlier experiments, we conclude that a combination of autoantibodies against multiple epitopes is necessary for the induction of glomerular damage in this model for lupus nephritis. PMID- 9394244 TI - The resurgence of selective contracting restrictions. AB - As managed care has spread, so has legislation to force plans to contract with any willing provider (AWP) and give patients freedom of choice (FOC). Managed care organizations' selective networks and provider integration reduce patient access to providers, along with provider access to paying patients, so many providers have lobbied for AWP-FOC laws. In opposition are managed care organizations (MCOs), which want full freedom to contract selectively to control prices and utilization. This article comprehensively describes laws in all fifty one jurisdictions, classifies their relative strength, and assesses the implications of the laws. Most are relatively weak forms and all are limited in application by ERISA and the federal HMO Act. The article also uses an associative multivariate analysis to relate the selective contracting environments to HMO penetration rates, rural population, physician density, and other variables. States with weak laws also have higher HMO penetration and higher physician density, but smaller rural populations. We conclude that the strongest laws overly restrict the management of care, to the likely detriment of cost control. But where market power is rapidly concentrating, not restricting selective contracting could diminish long-term competition and patient access to care. In the face of uncertainty about the impact of these laws, an intermediate approach may be better than all or nothing. States should consider mandating that plans offer point-of-service options, for a separate premium. This option expands patient choice of plans at the time of enrollment and of providers at the time of care, yet maintains plans' ability to control core providers. PMID- 9394245 TI - Medicaid managed care and the family planning free-choice exemption: beyond the freedom to choose. AB - Family planning services represent one of the most common managed care services, particularly in the case of Medicaid. In 1986, Congress enacted legislation exempting family planning services from mandatory managed care requirements. The effect of this legislation was to permit beneficiaries enrolled in mandatory managed care plans to continue to obtain family planning benefits from the providers of their choice, regardless of the providers' network status. The purpose of the law was to assure that managed care would not impede individuals' access to benefits. Subsequent experiences surrounding implementation of this provision indicated that the Health Care Financing Administration failed either to define the scope of the exemption or to provide guidance to states regarding the various issues that would have to be resolved in exempting certain primary care services from mandatory managed care. States, in turn, developed working definitions of exempt family planning services that omitted treatment for sexually transmitted diseases and also delegated to their managed care contractors the responsibility to design implementation and payment arrangements. This systematic failure to articulate the scope and functional elements of the exemption consequently led to nonpayment of providers, and ultimately, to denial of care in some cases. Moreover, plans and community providers alike relied on the exemption to justify their failure to come to grips with the requirements and challenges of managed care. We conclude that exempting primary care services from managed care requirements may raise as many questions as it answers, and instead recommend that states, plans, and community providers focus on developing managed care systems that are responsive to patient needs. PMID- 9394246 TI - Should health insurance cover IVF? Issues and options. AB - An emotional debate has attended the question of whether health insurance should cover the cost of in vitro fertilization (IVF) for infertile couples. Some private health plans have opted to cover IVF, although most have not. Ten states have mandated that it be included or offered as a standard benefit for private health insurance plans. This article analyzes several key issues in the debate: the impact of insurance coverage; the cost-effectiveness of IVF; valuing the benefit of IVF; and adoption as an alternative. It recommends policy action in several areas: more efficiently allocating resources for IVF (by giving priority to couples with better chances of success, and by making more extensive use of facilities with higher success rates); ensuring that clear and reliable information about the effectiveness of IVF is available; and leveling the playing field between IVF and adoption. PMID- 9394247 TI - Social science and the public agenda: reflections on the relation of knowledge to policy in the United States and abroad. AB - It is tempting to oversell the practical value of applied research. A hard look at the effects of U.S. social science on public policy in areas such as active labor market policies (training, job creation, placement, etc.), crime prevention, fiscal policy, poverty reduction, and health care reform suggests an inverse relationship between social science consensus and policy and budgetary decisions. Fragmented and decentralized political economies (e.g., the United States) foster policy segmentation and isolated, short-run single-issue research- often politicized and misleading. More corporatist democracies (such as Sweden, Norway, Austria, and Germany) evidence a tighter relation between knowledge and power in which a wider range of issues is connected, longer-range effects are sometimes considered, and research is more often actually used for planning and implementation. Even in less hospitable societies, however, social science does make its way in the long run. Favorable conditions and examples are discussed. PMID- 9394248 TI - Learning from others: shall the last be the first? AB - The cross-national exchange of ideas and experience in health care reform has, in recent years, reached epidemic proportions. Taking stock of this suggests the need for critical caution. The various participants in the process have different motives; models may be exported before they have been fully tested; information may be sought chiefly as political ammunition; and selective perception often distorts the evidence. A more helpful approach might be to concentrate on evaluating the experience of countries over time, rather than concentrating on the latest fashionable panacea, and to focus more on the process of introducing change. PMID- 9394250 TI - Institutions of reflective practice. PMID- 9394249 TI - Funding by the Center for Indoor Air Research (CIAR) PMID- 9394251 TI - Chitosan: properties, preparations and application to microparticulate systems. AB - Chitosan, a hydrophilic biopolymer, is obtained industrially by hydrolysing the aminoacetyl groups of chitin. It is a natural, non-toxic, biodegradable polysaccharide available as solution, flake, fine powder, bead and fibre. The sources, biochemical aspects, structure and chemical modification, physico chemical and functional properties, and applications of chitosan have been investigated extensively in the literature. In this paper, the attractive properties and broad applications of chitosan-based microparticles, their versatile properties, different preparation methods, and pharmaceutical and biopharmaceutical applications are reviewed. PMID- 9394252 TI - Comparison of two commercial brands of microcrystalline cellulose for extrusion spheronization. AB - Two commercial brands of microcrystalline cellulose, the widely used Avicel PH 101 and the newly available Pharmacel 101, were compared for aqueous extrusion spheronization of a model mix with lactose. Based on the results of multi-level experiments employing pellet size analysis by sieving and sphericity determination by image analysis, Avicel was shown to be less adversely affected by variation in added water or speed of spheronization. Physicochemical testing of powder samples from both brands was carried out using laser particle sized analysis, density determinations, differential scanning calorimetry and X-ray diffraction studies. The results indicated that the improved ease of processing with Avicel may be related to its smaller particle size with less aggregates, improved flow, lower depolymerization temperature range and absence of traces of cellulose II in its cellulose I content. PMID- 9394253 TI - Preparation of polymeric microspheres by the solvent evaporation method using sucrose stearate as a droplet stabilizer. AB - Polymeric microspheres containing nicardipine hydrochloride (HCl) as a reference drug were prepared with the acrylic polymers Eudragit RS and L by the solvent evaporation method. Different concentrations of sucrose stearate as a droplet stabilizer were used. Sucrose stearate affected the diffusion rate of the solvent from the preliminary emulsion droplets to the outer phase for the formation of microspheres. Increasing concentrations of sucrose stearate in the formulations caused increasing porosity on the surface of the microspheres. However, a correlation between the concentrations of sucrose stearate and diameters of microspheres could not be assessed. From this point of view, during processing, applied stirring rate was important. PMID- 9394254 TI - Development of vegetable extracts by microencapsulation. AB - The microencapsulation of essential oils offers protection against oxidation and evaporation, and allows the concurrent utilization of several vegetable extracts. Complex coacervation methods have been described for essential oils. Even though microencapsulation involves wrapping the essential oils in shells, some difficulties arise in the process of stabilizing the essential oils: oil may be lost by evaporation and partial dissolution in the water-gelatin phase and this will vary with the type of essential oil being encapsulated. In order to investigate the efficacy of the gelatin-polyphosphate methods we analysed their essential oil microcapsules peppermint and rosemary, in particular their granulometric size distribution, oil content (%) and encapsulation yield (%). In addition the essential oils were analysed by GC before and after microencapsulation so as to investigate the loss of their components during the process. PMID- 9394255 TI - In vitro characterization of a controlled-release chlorpheniramine maleate delivery system prepared by the air-suspension technique. AB - Non-pareil cores were spray-coated with a chlorpheniramine maleate (an alkylamine antihistamine) layer and a Eudragit NE30D overcoat in a Wurster air-suspension apparatus. In vitro dissolution studies demonstrated that drug release was a function of polymer membrane thickness. Polyethylene glycol 6000, as a hydrophillic additive, increased the in vitro release of chlorpheniramine maleate from the pellets. Pellets coated with 8.30% Eudragit NE30D, 0.50% talc and 1.00% polyethylene glycol 6000 were found to display desirable controlled release characteristics for chlorpheniramine maleate over the 8-h testing period, which were also comparable with that of Dykatuss capsules. The controlled release pellets exhibited first-order release characteristics for chlorpheniramine maleate. Reproducibility of the manufacturing conditions employed in the study were confirmed thus ensuring reproducibility of drug release characteristics between batches of chlorpheniramine maleate pellets. Drug release from the pellets was shown to be independent of the dissolution method and medium used. Pellets displayed no significant change in drug release characteristics relative to the initial drug release data when stored for 12 weeks at room temperature (20 +/- 2 degrees C) and for 8 weeks at a low temperature (5 +/- 1 degrees C). However, pellets stored at 37 degrees C with 80% relative humidity and at 40 +/- 2 degrees C showed a slower in vitro drug release after 8-week storage and therefore failed to maintain their initial drug release profile. PMID- 9394256 TI - Encapsulation of calcitonin in liposomes depends on the vesicle preparation method. AB - Calcitonin-loading was studied in liposomes composed of phosphatidylcholine, cholesterol and stearylamine in relation to the vesicle preparation method. Liposomes entrapping calcitonin were prepared by extrusion, sonication or from mixed micelles through the elimination of cholate by gel filtration. To understand the mode of calcitonin encapsulation in the vesicles, riboflavin was entrapped within the vesicles and taken as a simple model for the encapsulation of molecules in the aqueous phase. Interactions of calcitonin with the liposomal membranes were evaluated by studying the fixation of radiolabelled calcitonin to the outer surface of empty liposomes, and by preparing calcitonin-loaded LDL-like nanoparticles composed of phosphatidylcholine and cholesteryloleate. Calcitonin entrapment in the vesicles depends largely on the vesicle preparation method. When vesicles are prepared by removal of cholate from mixed micelles, relatively little calcitonin entrapment in the liposomes is obtained. In this type of vesicle, calcitonin is exclusively embedded in the vesicle bilayer. When vesicles are prepared by extrusion or sonication, calcitonin is found both in the aqueous and lipidic phases of the vesicles. Optimal calcitonin encapsulation was obtained when the liposomes were prepared by sonication. PMID- 9394257 TI - Microencapsulation of human insulin DEAE-dextran complex and the complex in liposomes by the emulsion non-solvent addition method. AB - Human insulin-DEAE (diethyl amino ethyl) dextran complex and human insulin DEAE dextran complex in liposomes were encapsulated in cellulose acetate butyrate (CAB) microcapsules by the emulsion non-solvent addition method. The ratio of core-to-coat used was 1:1. The average diameters of the complex microcapsules and the complex liposome microcapsules were 239.5 +/- 77.5 and 182.9 +/- 52.2 microns respectively. In vitro dissolution studies of both types of microcapsules in simulated intestinal fluid at pH 7.2 showed a sustained release of the complex and the complex liposome microcapsules with t50 = 1.5 h and 4 h respectively. This study can be applied to the further development of oral formulations of human insulin liposomes for diabetic treatment. PMID- 9394258 TI - Characterization of ciprofloxacin liposomes: derivative ultraviolet spectrophotometric determinations. AB - Neutral, (-) and (+) charged CF encapusulated liposome formulations prepared with EPC:Chol and DPPC:Chol lipids were investigated for their loading capacity, encapsulation % and release properties. CF amounts in the liposomes were estimated using derivative UV spectroscopy. Among the liposome formulations DPPC:Chol and EPC:Chol neutral formulations had a significantly higher loading capacity and slowest release rate compared with (-) and (+) charged liposomes. The derivative UV spectroscopy was found to be an easy and sensitive method for direct estimation of CF in liposomes. PMID- 9394259 TI - Stability of cyclosporine-loaded poly-sigma-caprolactone nanoparticles. AB - The aim was to evaluate the long-term stability of cyclosporin A-loaded nanoparticle suspensions, stored at 8 and 25 degrees C. The stability of freeze dried samples was also investigated. Nanoparticles (NP) of poly-sigma caprolactone (P sigma CL), a biodegradable polymer, were obtained by a modified nanoprecipitation method. A central composite experimental design was used to investigate the simultaneous effect of technological factors (temperature of the aqueous phase and needle gauge) and formulation variables (volume of acetone and the amount of polymer and surfactant). The effect of these variables on the stability of the 100-220 nm particles obtained was evaluated. The percentage of cyclosporin A (CyA) encapsulated in the NP suspensions stored at 8 and 25 degrees C for at least 3 months remained unaltered. Moreover, there was no change in the size of NP. After 4 months storage, the physical stability of the preparation was affected. NP aggregates could be observed by light microscopy. Reconstituted freeze-dried preparations showed a mean increase of 1% in the incorporated drug and also a considerable increase in mean size and size distribution. Additional experiments investigated the effect of freezing temperature (-70 and -196 degrees C) and of 5, 10 and 20% (w/v) cryoprotector (mannitol, sorbitol, glucose and threalose) on 100 nm particles. The addition of glucose and threalose at concentrations > 10% permitted adequate reconstitution of the freeze-dried product with conservation of the encapsulated CyA. PMID- 9394260 TI - Depression and coronary heart disease: observations and questions. AB - The evidence that depressive symptomatology precedes the onset of the acute coronary syndromes and influences the course of disease after their manifestation is accumulating. However, we still are far short of proof that depression has a causal role in the etiology and pathogenesis of coronary heart disease (CHD). Some unsolved questions concern the causes and the nature of the depression preceding a first or recurrent cardiac event, the biological mechanisms relating depression and CHD, the time window of the exposure-disease association, and the power of therapy programs for depression to reduce the risk of a first or recurrent cardiac event. PMID- 9394261 TI - The psychological and behavioral management of angina. PMID- 9394262 TI - Improving the effectiveness of routine care for somatization. PMID- 9394263 TI - Liaison psychiatry and psychology in dentistry. AB - Dentists are trained to provide treatment for patients with straightforward problems that respond to routine therapy and do not recur. However, patients may present to dentists and complain solely of physical symptoms such as toothache, headache, and facial pain: only after much inappropriate treatment these symptoms are revealed to be due to emotional disturbance. The dentist may spend hours investigating such patients, in some of whom dental pathology may be present, but the symptoms and ensuing disability cannot be satisfactorily explained as a result. There are other patients who are preoccupied by physical symptoms or by their appearance. In others, anxiety may manifest itself as a phobia, or a dysmorphic concern about certain aspects of their appearance. This article reviews the role of liaison psychiatry and psychology in dentistry. PMID- 9394264 TI - Predictive genetic testing: psychological factors. AB - Advances in medical technology such as those linked to the human genome project are increasing the potential for predictive testing for a wide range of health threats. There have not been comparable advances in understanding of the psychological factors involved in such testing. These factors and issues relating to them are examined, and it is suggested that a cognitive-behavioral approach to the understanding and management of adverse reactions to testing is likely to be particularly fruitful. The use of such an approach should result in the development of effective pre- and posttest interventions to prevent, minimize, and manage distress associated with screening. PMID- 9394265 TI - Neurovegetative dystonia--psychiatric evaluation of 40 patients diagnosed by general physicians in Brazil. AB - The diagnosis of neurovegetative dystonia (NVD) is commonly made by general physicians in Brazil, but its precise meaning is unclear. Anecdotal evidence suggests that it is used to describe patients with a wide range of psychological and physical symptoms and is often used pejoratively, in a similar way to "crocks" in the USA. Forty patients who had been diagnosed as having NVD by general physicians working in a triage department of a general public hospital were compared with 40 non-NVD patients, matched for age and gender, from the same department. Patients were evaluated by a psychiatrist who was blind to the diagnosis that had been made. The assessment included a structured sociodemographic questionnaire, the Clinical Interview Schedule (CIS), and a routine psychiatric interview using DSM-III-R criteria. Using the CIS, the "reported symptoms" that most distinguished NVD patients from controls were somatic and anxiety, whereas for "manifest abnormality" NVD patients displayed more anxiety, histrionic behavior, hypochondriasis, and depressive thoughts. A total of 92.5% of NVD patients received diagnoses using DSM-III-R criteria compared to 37.5% of controls. The relative risk of NVD patients subsequently receiving a psychiatric disorder was 8.3 (95% CI = 2.5-43.1, p < 0.001). Although general physicians correctly identify most patients with psychiatric disorder they miss many others. Furthermore, they use an obsolete diagnostic category which has no psychiatric currency. Medical students and residents need better psychiatric training so that they can correctly identify patients in general medical settings who are suffering from mental disorders and make a diagnosis using accepted psychiatric terminology. PMID- 9394266 TI - Medication misuse, abuse and dependence in chronic pain patients. AB - We report the prevalence of drug use, misuse, abuse, and dependence in 125 chronic pain patients attending specialist pain clinics in South London. A total of 110 patients (88%) were taking medications for their pain problem. Opioid analgesics (69.6%), nonopioids (48%), antidepressants (25%), and benzodiazepines (17.6%) were the drugs most frequently used. Psychoactive substance abuse or dependence (DSM-III-R) was diagnosed in 12%. A total of 9.6% of the patients met the DSM-III-R criteria for substance abuse or dependence in remission. Data are also presented on the misuse and abuse of nonpsychoactive drugs, qualitative information on how patients use drugs, and the information they have received about medication. PMID- 9394267 TI - Personality change following head injury: assessment with the NEO Five-Factor Inventory. AB - We evaluated personality change following head injury in 68 patients at 6 months postinjury using the NEO Five-Factor Inventory to assess the five personality dimensions of the Five-Factor Model of Personality. All items had to be rated twice, once for the preinjury and once for the current status. Twenty-eight trauma patients with injuries to other parts of the body than the head were used as controls. For the head-injured group, 63 relatives also completed the questionnaire. The results showed no differences between the ratings of head injured patients and the ratings of trauma control patients. Both groups showed significant change in the personality dimensions Neuroticism, Extraversion, and Conscientiousness. Compared to their relatives, head-injured patients report a smaller change in Extraversion and Conscientiousness. Changes were not reported on the Openness and Agreeableness scales, by neither the head-injured or their relatives, nor by the trauma controls. PMID- 9394268 TI - Gender, self-reported depressive symptoms, and sleep disturbance among older community-dwelling persons. FICSIT group. Frailty and Injuries: Cooperative Studies of Intervention Techniques. AB - The purposes of this report are: (1) to investigate the association between sleep disturbances and depressive symptomatology in older adults; (2) to evaluate the degree to which gender serves to mediate this relationship; and (3) to determine whether several predefined covariates help to explain the association between sleep disturbance and depressive symptoms. This is a retrospective and cross sectional analysis of baseline data from 485 elderly adults enrolled in three of the eight clinical sites participating in the Frailty and Injuries: Cooperative Studies of Intervention Techniques (FICSIT) trials. FICSIT was a linked series of randomized clinical trials which evaluated the impact of various exercise interventions on several measures of frailty in older adults. Women reported more depressive symptoms and more sleep disturbances than men. Sleep disturbances were independently associated with depressive symptoms, bodily pain, a history of falling, limited education, being married, and being female. Gender interactions suggest that, although women reported more depressive symptoms and more chronic health conditions than men, both may be more important predictors of sleep disturbance in men. By contrast, being married may be more predictive in women. Finally, the data suggest a stronger relationship between sleep disturbance and depressive symptoms in men than in women. PMID- 9394270 TI - Negative affect and the seeking of medical care in university students with irritable bowel syndrome: a preliminary study. AB - In a preliminary study using only self-report measures, university students completed questionnaires about their bowel symptoms and trait anxiety. Results showed that students with irritable bowel syndrome (IBS) reported higher trait anxiety than asymptomatic controls. Among the students with IBS, there were no significant differences in trait anxiety between those who had sought medical care for IBS mostly from a primary care physician, and those who had not sought care for IBS. Students who had sought medical care for IBS reported being more bothered by the symptoms and were more concerned about their meaning than those students who had not sought care. The results are compared to other research with IBS patients referred to specialist clinics, and a distinction is made between initial vs. continued care seeking for IBS. PMID- 9394269 TI - Acute phase proteins in major depression. AB - Extensive evidence exists associating depression with changes in the immune system. The present study evaluates the levels of complement components C3 and C4, C-reactive proteins, and IL-6 in patients who met DSM-III-R diagnostic criteria for major depressive disorder, as well as controls. Whereas no significant differences between the mean levels of C3 could be detected between depressed patients and controls, the levels of C4, IL-6 (where detected), and C reactive protein were significantly raised in the group with a depressive disorder. Our study suggests an interaction between psychological state and immune systems operative in host defenses. PMID- 9394271 TI - The comorbidity, relationship and treatment implications of borderline personality and obesity. AB - Studies indicate that a significant minority of obese individuals in clinical studies meet criteria for borderline personality. Although the relationship between obesity and borderline personality remains unexplained, the following article discusses the implications of treating obesity among individuals with this personality disorder. Longitudinal intervention, normalizing or regulating eating patterns, and reframing weight plateaus are emphasized. PMID- 9394272 TI - An accurate method for computer-generating tungsten anode x-ray spectra from 30 to 140 kV. AB - A tungsten anode spectral model using interpolating polynomials (TASMIP) was used to compute x-ray spectra at 1 keV intervals over the range from 30 kV to 140 kV. The TASMIP is not semi-empirical and uses no physical assumptions regarding x-ray production, but rather interpolates measured constant potential x-ray spectra published by Fewell et al. [Handbook of Computed Tomography X-ray Spectra (U.S. Government Printing Office, Washington, D.C., 1981)]. X-ray output measurements (mR/mAs measured at 1 m) were made on a calibrated constant potential generator in our laboratory from 50 kV to 124 kV, and with 0-5 mm added aluminum filtration. The Fewell spectra were slightly modified (numerically hardened) and normalized based on the attenuation and output characteristics of a constant potential generator and metal-insert x-ray tube in our laboratory. Then, using the modified Fewell spectra of different kVs, the photon fluence phi at each 1 keV energy bin (E) over energies from 10 keV to 140 keV was characterized using polynomial functions of the form phi (E) = a0[E] + a1[E] kV + a2[E] kV2 + ... + a(n)[E] kVn. A total of 131 polynomial functions were used to calculate accurate x-ray spectra, each function requiring between two and four terms. The resulting TASMIP algorithm produced x-ray spectra that match both the quality and quantity characteristics of the x-ray system in our laboratory. For photon fluences above 10% of the peak fluence in the spectrum, the average percent difference (and standard deviation) between the modified Fewell spectra and the TASMIP photon fluence was -1.43% (3.8%) for the 50 kV spectrum, -0.89% (1.37%) for the 70 kV spectrum, and for the 80, 90, 100, 110, 120, 130 and 140 kV spectra, the mean differences between spectra were all less than 0.20% and the standard deviations were less than approximately 1.1%. The model was also extended to include the effects of generator-induced kV ripple. Finally, the x-ray photon fluence in the units of photons/mm2 per mR was calculated as a function of HVL, kV, and ripple factor, for various (water-equivalent) patient thicknesses (0, 10, 20, and 30 cm). These values may be useful for computing the detective quantum efficiency, DQE(f), of x-ray detector systems. The TASMIP algorithm and ancillary data are made available on line at http:/(/)www.aip.org/epaps/epaps.html. PMID- 9394273 TI - Three-dimensional reconstruction of curves from pairs of projection views in the presence of error. I. Algorithms. AB - We have previously described an approach to 3D intracerebral vascular reconstruction that uses an MRA as a reconstruction base. Additional vessels seen only by angiography are added by segmenting 2D curves from projection angiograms and reconstructing these curves into 3D, building upon the MRA. Intracerebral vascular reconstruction is difficult for at least two reasons. First, 2D curves must be associated on projection images even when the human eye cannot do so. Second, 3D curves must be reconstructed in the presence of errors such as misregistration, image distortion, and misdefinition of 2D curves. This paper is the first of two that address the specific issue of reconstruction of a 3D curve from a given pair of 2D curves in the presence of error. The method explicitly separates what can and cannot be determined from a pair of projection views. It is also capable of recognizing interruptions produced by viewplane errors, of continuing reconstruction beyond such interruptions, and of localizing and estimating the magnitude of the interruptions. These measurements can also be used to estimate the lengths of regional disparities between a pair of 2D curves, leading to a quantitative estimate of the capacity of a pair of 2D curves to combine to create a 3D object (match value). Match values can be used, in turn, as part of the strategy for automatically associating pairs of 2D curves. This paper provides methods for reconstructing a given pair of 2D curves into 3D in the presence of error and for calculating match values. Error analysis is given in the companion report. PMID- 9394274 TI - Three-dimensional reconstruction of curves from pairs of projection views in the presence of error. II. Analysis of error. AB - We have previously described an approach to 3D intracerebral vascular reconstruction that uses an MRA as a reconstruction base. Additional vessels seen only by angiography are added by segmenting 2D curves from projection angiograms and reconstructing these curves into 3D, building upon the MRA. This paper is the second of two that discuss the specific problem of reconstructing a 3D curve from a given pair of 2D curves in the presence of error. The method presented is capable of detecting and handling many errors produced by misregistration, image distortion, or misdefinition of 2D curves. The first paper gives an algorithm. The current paper discusses factors affecting the accuracy of a reconstructed curve, with emphasis upon registration error. We analyze the spatial accuracy of a reconstructed point in terms of the relationships between pixel size, relative viewing angle, 3D point location, and registration error. We provide a theoretical framework that, given the known error properties of a registration algorithm, allows optimization of the viewing geometry so as to produce the highest precision of point reconstruction. A major focus is the effect of registration error upon the reconstruction of a curve. We subdivide registration error into two types, one of which produces smoothly continuous point placement errors and the other of which produces pixel pairing errors. We test our ability to reconstruct a 3D curve in the presence of both. Finally, we summarize approaches to other sources of error. We conclude with a list of recommendations to optimize reconstruction accuracy. When projection points are associated by the rules of epipolar geometry, viewplane point displacements should not exceed 1.5-2 mm along the axis perpendicular to epipolar planes. PMID- 9394275 TI - Computer simulation of time-gated transillumination and reflection of biological tissues and tissuelike phantoms. AB - A simulation technique is employed to explore the possibility of locating millimeter-sized objects, immersed in turbid media, from time-gated measurements of the transmitted or reflected light. The simulation results for tissuelike phantoms are compared to experimental transillumination data and excellent agreement is found. Simulations of time-gated reflection experiments show that it is possible to detect objects of 1 mm diameter. This may open new possibilities for medical diagnosis of breast cancer in an early stage. PMID- 9394276 TI - A method for quantifying SPECT uniformity. AB - Field uniformity is an important parameter for monitoring the performance of SPECT imaging systems. However, it is difficult to apply objective measures of uniformity because of the large variance associated with reconstructed images. In the proposed method, annular sampling of the SPECT uniformity image is used to reduce the noise level without decreasing the magnitude of uniformity artifacts. The reconstructed uniformity image is sectioned into annular rings centered on the center of rotation to match the expected distribution of uniformity artifacts. Statistical fluctuations are reduced by averaging the counts within the annular rings, allowing the use of objective measures of field uniformity such as integral uniformity. Application of the annular sampling technique on simulated and phantom uniformity images showed that the technique could reliably quantify SPECT uniformity artifacts at acceptable count levels. As a result this method can be used to objectively evaluate SPECT field uniformity in systems which utilize parallel collimation and circular orbits. PMID- 9394277 TI - The effect of gamma ray penetration on angle-dependent sensitivity for pinhole collimation in nuclear medicine. AB - The sensitivity of a pinhole collimator for gamma ray imaging in nuclear medicine is dependent on the angle of incidence of the gamma rays. The effect of penetration near the pinhole aperture on angle-dependent sensitivity was investigated using experimental measurements and numerical modeling. Projection data measurements were acquired with Tc-99m and I-131 point sources using tungsten pinhole inserts with 1.0 to 4.0 mm diameter apertures. Curves of the form sinx theta, where theta is the angle of the incident ray with the surface of the detector crystal, were fit to sensitivity measurements from the projection data. Experimentally measured x values were between 3.3 and 4.1 for Tc-99m and between 5.1 and 7.2 for I-131. Penetration near the pinhole aperture was modeled using (1) an expression for effective pinhole diameter that is a generalization of Anger's formula for normally incident photons and (2) a photon transport simulation code. Experimentally measured sensitivity exponents x from new and previously reported experimental observations were modeled within 15% by the numerical simulations. For modeling using the generalized expression for effective diameter the average error was 1.4% and the standard deviation was 7.7%. For the photon transport simulation code the average error was 1.5% and the standard deviation also was 7.7%. The effect of pinhole aperture design parameters on angle-dependent sensitivity for high resolution pinhole apertures was modeled using a photon transport simulation code. The sensitivity exponents x were greater for 364 keV photons than for 140 keV photons and were greater for small aperture diameters, small acceptance angles, and large aperture channel heights. These results provide theoretical justification for incorporating sinx theta sensitivity corrections, with x greater than the value of 3 for an impenetrable pinhole, in filtered back projection and iterative reconstruction algorithms for single photon emission computed tomography (SPECT) pinhole imaging. Simulated I-131 SPECT studies for uniformly active cylinders showed that activity concentrations were underestimated toward the outside of the cylinders when a sin3 theta rather than the correct sinx theta sensitivity correction was applied in image reconstruction. PMID- 9394278 TI - Radiation doses in radiation therapy are not safe. PMID- 9394279 TI - Calculating dose and output factors for wedged photon radiotherapy fields using a convolution/superposition method. AB - We have developed a convolution/superposition method to calculate dose distributions in photon treatment fields with beam modifiers such as physical wedges. The dose component due to wedge generated radiation was accounted for by using an extended phantom model, which integrated a wedge, an air gap, and a patient phantom as the calculation phantom. The inhomogeneities in the extended phantom and the effect of beam hardening by the wedge were both corrected for in the convolution dose calculation. The calculated dose was verified by Monte Carlo simulation of the same extended phantom. A new dual photon source model was also used in the convolution method to account for both primary photons from the target and extra-focal photons from the primary collimator and flattening filter. Thus, realistic photon energy fluence distributions in the extended phantom were used for the dose calculation. The calculated dose distributions and the wedge factors agreed with the measured data within 2% for a variety of treatment fields including asymmetric fields. Our results showed that the wedge-generated radiation could contribute a significant fraction of the total dose in patients. This dose component depends on a specific field configuration, thus wedge factor changes with photon energy, wedge angle, field size, depth, and patient phantom SSD. The variation of the wedge factor can be predicted accurately by our convolution approach with the extended phantom model, which allows for more accurate dose or monitor unit computation for photon fields with beam modifiers. PMID- 9394280 TI - Correcting kernel tilting and hardening in convolution/superposition dose calculations for clinical divergent and polychromatic photon beams. AB - To account for clinical divergent and polychromatic photon beams, we have developed kernel tilting and kernel hardening correction methods for convolution dose calculation algorithms. The new correction methods were validated by Monte Carlo simulation. The accuracy and computation time of the our kernel tilting and kernel hardening correction methods were also compared to the existing approaches including terma divergence correction, dose divergence correction methods, and the effective mean kernel method with no kernel hardening correction. Treatment fields of 10 x 10-40 x 40 cm2 (field size at source to axis distance (SAD)) with source to source distances (SSDs) of 60, 80, and 100 cm, and photon energies of 6, 10, and 18 MV have been studied. Our results showed that based on the relative dose errors at a depth of 15 cm along the central axis, the terma divergence correction may be used for fields smaller than 10 x 10 cm2 with a SSD larger than 80 cm; the dose divergence correction with an additional kernel hardening correction can reduce dose error and may be more applicable than the terma divergence correction. For both these methods, the dose error increased linearly with the depth in the phantom; the 90% isodose lines at the depth of 15 cm were shifted by about 2%-5% of the field width due to significant underestimation of the penumbra dose. The kernel hardening effect was less prominent than the kernel tilting effect for clinical photon beams. The dose error by using nonhardening corrected kernel is less than 2.0% at a depth of 15 cm along the central axis, yet it increased with a smaller field size and lower photon energy. The kernel hardening correction could be more important to compute dose in the fields with beam modifiers such as wedges when beam hardening is more significant. The kernel tilting correction and kernel hardening correction increased computation time by about 3 times, and 0.5-1 times, respectively. This can be justified by more accurate dose calculations for the majority of clinical treatments. PMID- 9394281 TI - Decision theoretic steering and genetic algorithm optimization: application to stereotactic radiosurgery treatment planning. AB - Treatment planning for stereotactic radiosurgery and fractionated radiotherapy is currently a labor intensive, operator-dependent process. Many degrees of freedom exist to make rigorous optimization intractable except by computationally intelligent techniques. The quality of a given plan is determined by an aggregate of clinical objectives, most of which are subject to competing tradeoffs. In this work, we present an autonomous scheme that couples decision theoretic guidance with a genetic algorithm for optimization. Ordinal ranking among a population of viable treatment plans is based on a generalized distance metric, which promotes a decreasing hyperfrontier of the efficient solution set. The solution set is driven toward efficiency by the genetic algorithm, which uses the tournament selection mechanism based on the ordinal ranking. Goals and satisficing conditions can be defined to signal the ultimate and the minimum achievement levels in a given objective. A conventionally challenging case in radiosurgery was used to demonstrate the practical utility and the problem-solving power of the decision theoretic genetic algorithm. Treatment plans with one isocenter and four isocenters were derived under the autonomous scheme and compared to the actual treatment plan manually optimized by the expert planner. Quality assessment based on dose-volume histograms and normal tissue complication probabilities suggested that computational optimization could be driven to offer varying degrees of dosimetric improvement over a human-guided optimization effort. Furthermore, it was possible to achieve a high degree of isodose conformity to the target volume in computational optimization by increasing the degree of freedom in the treatment parameters. The time taken to derive an efficient planning solution was comparable and usually shorter than in the manual planning process, and can be scaled down almost linearly with the number of processors. Overall, the autonomous genetic algorithm scheme was found to be powerful and versatile as a computationally intelligent counterpart to human guided strategies in treatment optimization for stereotactic radiosurgery and radiotherapy. PMID- 9394282 TI - Inverse radiation treatment planning using the Dynamically Penalized Likelihood method. AB - In this paper we present a new method of solving the inverse radiation treatment planning problem. The method is based on a Maximum Likelihood Estimator with dynamically changing penalization terms. The resulting Dynamically Penalized Likelihood (DPL) algorithm achieves a dose distribution of excellent uniformity in a tumor volume and a much lower dose in regions containing sensitive volumes. A simple model of a patient and of energy deposition has been used for the initial results presented: a two-dimensional computer generated phantom and monochromatic x rays, without scattering. Three two-dimensional problems are solved with the DPL algorithm, corresponding to different size and spatial relationships between the tumor and sensitive tissue volumes. The results show that the DPL algorithm is robust and flexible; it only requires moderate computation times and leads to promising solutions, even in rather difficult problems. The results encourage the extension of the present work to more realistic therapy situations. PMID- 9394283 TI - A sector-integration method for calculating the output factors of irregularly shaped electron fields. AB - An empirical model is presented that uses a sector-integration method for calculating the output factors of irregularly shaped electron fields. The sector integration method accounts for changes in electron fluence, lateral scatter equilibrium, and scatter from the edge of a cutout shield. This method is tested for elliptical and rectangular fields with a ratio of the long-to-short axis as great as 4 to 1. Differences between measured and calculated values for output factors were less than +/- 1%. Comparisons were also carried out for a large number of cutout shields that were used in the clinic and similar levels of accuracy were obtained. PMID- 9394284 TI - An equivalent square field formula for determining head scatter factors of rectangular fields. AB - A simple formula is derived for the calculation of an equivalent square field that gives the same head scatter factor as a given rectangular field. This formula is based strictly on the configuration of a medical linear accelerator treatment head. The geometric parameters used are the distances between the target and the top of each field-defining aperture. The formula accounts for both the effect of field elongation and the collimator exchange effect. This method predicts the output to within 1% accuracy for both open and wedged fields and does not require any new measured data other than the field size dependence of head scatter for a range of square field sizes. Interestingly, the formula we derived has the same format as the formula that was empirically obtained by Vadash and Bjarngard [Med. Phys. 20, 733-734 (1993)]. PMID- 9394285 TI - An approximation of central-axis absorbed dose in narrow photon beams. AB - In narrow photon beams of therapeutic energy range, the absorbed dose derived from experimental measurements is subject to a significant error. The error stems from high dose gradients characteristic to small radiation fields and from finite probe dimensions. In this study, a simple model for the narrow-beam absorbed dose is described. It is shown that broad-beam dose data are sufficient to predict a narrow-beam dose. The dose is calculated as a sum of primary and scatter components given in the form of respective analytical functions. For both functions, numerical coefficients are determined in broad-beam geometry. The model is evaluated by comparing calculated dose values with the Monte Carlo simulated narrow-beam dose data for 6 and 15 MV x rays. PMID- 9394286 TI - Forward dose perturbation at high atomic number interfaces in kilovoltage x-ray beams. AB - High atomic number (Z) materials such as lead, used for field shaping and shielding normal tissues in kilovoltage beams could produce significant dose enhancement in the forward direction contrary to our normal belief with respect to the attenuation of photon beams. Such a dose enhancement has not been studied in kilovoltage beams, which is investigated in this study. Using a Siemens ortho voltage unit (60-240 kVp) and a thin window (5 microns) parallel plate ion chamber, forward dose perturbation factor (FDPF) was measured at interfaces created by high- and low-Z materials. The FDPF is defined as the ratio of doses with and without an interface (FDPF = Di/Dh; where Di is the dose at an interface and Dh is the dose in a homogeneous medium). Results indicate that dose enhancement (FDPF > 1) as high as 20-fold can be observed for a thin (> or = 0.02 mm) Pb sheet in contact with soft tissue. The magnitude of FDPF is relatively independent of field size and falls off exponentially with Pb thickness. The typical photon beam attenuation takes at a thickness > 1 mm. This intense dose enhancement is localized within 250 microns of the interface. The FDPF is energy dependent but saturates above 140 kVp, unlike the backscatter dose perturbation that peaks around 200 kVp. The FDPF varies inversely with the thickness of high Z and distance between the surface and high-Z medium. The FDPF falls off rapidly to a level of photon transmission usually predicted by exponential attenuation when distance is increased. In conclusion, with kilovoltage beam, a high-Z medium placed in contact with soft tissue may not attenuate radiation dose unless adequate thickness and proper distance between the surface and high-Z medium is used. The localized intense dose enhancement (approximately 20-fold) created by the high-Z interface could be exploited for clinical use. PMID- 9394287 TI - The investigation of 32P wire for catheter-based endovascular irradiation. AB - The dose distribution from a 32P source has been measured and calculated in order to evaluate its application in endovascular irradiation. The source dimension was 27 mm in length and 0.3 mm in diameter and was embedded in the end of a Ni-Ti wire. Dose measurements were performed using radiochromic film in several specially designed tissue equivalent phantoms. Loevinger's point dose kernel was used for the calculation. The approximate dose rate at a radial distance of 1.5 mm from the center of the source was found to be 6.75 cGy/s per GBq (0.25 cGy/s per mCi), which allows the delivery of a therapeutic dose in a short time interval with a satisfactory homogeneity without stepping the source. However, the dose rate falls off almost exponentially along the radial distance. Therefore it may not be suitable for treating large diameter vessel from a centrally located source. The effect of a curved 32P wire source on the radial dose distribution was also investigated. The results showed that for a maximum bend of 180 degrees the dose rate was increased by as much as 20% along the inner radial distance but decreased by as much as 20% along the outer radial distance compared to the dose along a straight wire. However, for curvatures normally encountered in a clinical situation, the dose rate was changed less than 5%. PMID- 9394288 TI - MarCell software for modeling bone marrow radiation cell kinetics. AB - Differential equations were used to model cellular injury, repair, and compensatory proliferation in the irradiated bone marrow. Recently, that model was implemented as MarCell, a user-friendly MS-DOS computer program that allows users from a variety of technical disciplines to evaluate complex radiation exposure. The software allows menu selections for different sources of ionizing radiation. Choices for cell lineages include progenitor, stroma, and malignant, and the available species include mouse, rat, dog, sheep, swine, burro, and man. An attractive feature is that any protracted irradiation can be compared with an equivalent prompt dose (EPD) in terms of cell kinetics for either the source used or for a reference such as 250 kVp x rays or 60Co. EPD is used to mean a dose rate for which no meaningful biological recovery occurs during the period of irradiation. For human as species, output from MarCell includes: risk of 30-day mortality; risk of whole-body cancer and leukemia based either on radiation induced cytopenia or compensatory cell proliferation; cell survival and repopulation plots as functions of time or dose; and 4-week recovery following treatment. PMID- 9394289 TI - A method for measuring the ionization fraction due to the chamber wall (alpha) and assessing its characteristics. AB - To calibrate a megavoltage therapy beam using an ionization chamber, it is necessary to know the fraction of the ionization arising in the chamber wall when this is made of a material different than the medium. A method for measuring the ionization fraction produced by electrons arising in the chamber wall (alpha) is presented here. The method uses three measurements at the same point in a medium in order to calculate alpha. These measurements are made using the examined chamber with and without a buildup cap and one reference chamber of wall material equivalent to the medium (i.e., in our case, A1 and A-150 were used as wall materials for the examined and the reference chamber, respectively). Using this method, it is possible to calculate alpha in the medium for a series of irradiation conditions and assess its characteristics. Two main conclusions came out of this assessment. The first one is the independence of alpha from the wall material, even if this is aluminum (alpha is only dependent on wall thickness expressed in g cm-2). The second one is that alpha depends on the irradiation conditions; it increases with field size and depth. PMID- 9394290 TI - What studies of fusion peptides tell us about viral envelope glycoprotein mediated membrane fusion (review). AB - This review describes the numerous and innovative methods used to study the structure and function of viral fusion peptides. The systems studied include both intact fusion proteins and synthetic peptides interacting with model membranes. The strategies and methods include dissecting the fusion process into intermediate stages, comparing the effects of sequence mutations, electrophysiological patch clamp methods, hydrophobic photolabelling, video microscopy of the redistribution of both aqueous and lipophilic fluorescent probes between cells, standard optical spectroscopy of peptides in solution (circular dichroism and fluorescence) and attenuated total reflection-Fourier transform infrared spectroscopy of peptides bound to planar bilayers. Although the goal of a detailed picture of the fusion pore has not been achieved for any of the intermediate stages, important properties useful for constraining the development of models are emerging. For example, the presence of alpha-helical structure in at least part of the fusion peptide is strongly correlated with activity; whereas, beta-structure tends to be less prevalent, associated with non native experimental conditions, and more related to vesicle aggregation than fusion. The specific angle of insertion of the peptides into the membrane plane is also found to be an important characteristic for the fusion process. A shallow penetration, extending only to the central aliphatic core region, is likely responsible for the destabilization of the lipids required for coalescence of the apposing membranes and fusion. The functional role of the fusion peptides (which tend to be either nonpolar or aliphatic) is then to bind to and dehydrate the outer bilayers at a localized site; and thus reduce the energy barrier for the formation of highly curved, lipidic 'stalk' intermediates. In addition, the importance of the formation of specific, 'higher-order' fusion peptide complexes has also been shown. Recent crystallographic structures of core domains of two more fusion proteins (in addition to influenza haemagglutinin) has greatly facilitated the development of prototypic models of the fusion site. This latter effort will undoubtedly benefit from the insights and constraints gained from the studies of fusion peptides. PMID- 9394291 TI - A novel family of channel-forming, autotransporting, bacterial virulence factors. AB - Pathogenic bacteria produce virulence factors that cross the bacterial cell envelope from the cytoplasm to the extracellular milieu where they promote disease. The mechanisms of their export are poorly understood. We here characterize a family of autotransporter (AT) protein domains present at the C termini of several nonhomologous Gram-negative bacterial virulence factors. The family consist of 18 sequenced protein domains, the functionally characterized members of which catalyze export of (1) proteases, (2) virulence-related cell adhesins, (3) mediators of actin-promoted bacterial motility, (4) cytotoxins and (5) tissue invasion proteins. We (1) establish that these AT domains are homologous, (2) multiply align their sequences, (3) derive an AT family-specific signature sequence, and (4) define the evolutionary relationships between members of the family. Secondary structural predictions as well as average hydropathy, average similarity and average amphipathicity plots have allowed us to propose a specific 14 beta-stranded barrel structural model that may be applicable to all protein members of the AT family. We suggest that the AT domains became associated with active virulence factor domains by interdomain fusion events that occurred during the evolution of these complex proteins. PMID- 9394293 TI - Assignment of the human serotonin 1F receptor gene (HTR1F) to the short arm of chromosome 3 (3p13-p14.1). AB - In the present study, we report the chromosomal localization of the human 5-HT1F receptor gene (HTR1F) by the analysis of somatic cell hybrids. Based upon the HTR1F cDNA sequence, a primer set that reacted with human genomic DNA but not mouse or hamster genomic DNA was derived from the relatively nonconserved 5' untranslated and coding region. Using monochromosomal hybrid cell lines of the NIGMS Mapping Panel 2 we localized the HTR1F to human chromosome 3. To confirm the localization on chromosome 3 and to further sublocalize the HTR1F gene, a set of human cell hybrids regionally separating chromosome 3 into 7 regions was similarly analysed. Analysis of this regional panel showed that the HTR1F gene was located proximal to the 3p14.1 breakpoint in hybrid APH14 and distal to the breakpoint in 3p13 in hybrid APH13. This localizes the HTR1F gene to human chromosome 3p13-p14.1. PMID- 9394292 TI - Phosphatidylserine exposure and aminophospholipid translocase activity in Rh deficient erythrocytes. AB - Endogenous phosphatidylserine (PS) exposure and lipid transport activity have been investigated for seven unrelated cases of Rhnull erythrocytes. Endogenous PS exposure was measured by prothrombinase activity. Out of six cases studied, two Rhnull samples exhibited abnormal aminophospholipid exposure, as suggested by the measurement of a lower Km of factor Xa for prothrombin. Aminophospholipid translocase activity was measured through the transbilayer redistribution of spin labelled analogues of phospholipids. Provided that incubation conditions allow the maintainance of intracellular ATP level, no difference was observed between Rhnull and control erythrocytes, clearly indicating that the aminophospholipid translocase and Rh polypeptides are different molecular species. PMID- 9394294 TI - Aggregation, fusion and aqueous content release from liposomes induced by lysozyme derivatives: effect on the lytic activity. AB - Chemically modified lysozymes, namely: N-succinyl lysozyme, glycine methyl ester of N-succinyl lysozyme and oxoindole lysozyme have been prepared. Aggregation, fusion and leakage of phospholipid vesicles induced by these derivatives have been studied in comparison with the effect of the unmodified protein. The experiments were carried out with negatively charges 9PC/PA, 9:1) and uncharged (PC and PC/DOPE/Chol (10:5:5)) lipid vesicles of different packing. Fusion and aggregation of negatively charged phospholipid vesicles in induced by proteins positively charged at pH 7.0 involving electrostatic interactions, a similar pattern on fusion and aggregation of the least stably packed lipid vesicles points also to hydrophobic forces playing a role in the lipid-protein interaction. A conformational change of the protein involved increasing beta turns, loops and unordered structure at the expenses of beta-sheet without affecting alpha helix content. The conformational effect is necessary to provoke the effects studied, since one of the derivatives (N-succinyl lysozyme) neither changes conformation nor causes aggregation and fusion of vesicles. However, there is no relationship between lysozyme activity and fusion or aggregation of lipid vesicles that catalytic and fusogenci sites of, indicating lysozyme are topographically different. PMID- 9394295 TI - Membrane proteins at the interface of erythrocytes fused by treatment with polyethylene glycol. AB - Fusion of human red cells through the action of polyethylene glycol gives rise to pairs or higher clusters with a common membrane envelope, in which a barrier at the position of the original interface can be seen in phase contrast. At early times this septum contains lipids, as judged by labelling with a fluorescent lipophile, and transmembrane protein; this was shown by the presence of the preponderant component, band 3, detected by a fluorescent label, covalently attached before fusion at an extracellular site, or by immunofluorescence with anti-band 3 antibody. Ankyrin, which is bound to band 3, is also observed in the septum. The lipid thereafter disappears from the interface, carrying most of the band 3 with it. A continuous membrane skeletal network, defined by the presence of spectrin (detected by immunofluorescent staining in epifluorescence and confocal microscopy) appears to persist for long periods, but in many cells interruptions develop in the septum. In other fused pairs, particularly at longer times, the interface is seen to have vanished completely. Protease inhibitors have no discernible effect on any of these observations. The results suggest a model for the events that follow fusion. Covalent cross-linking of membrane proteins beyond a critical level causes inhibition of fusion, suggesting that proteins, probably the membrane skeletal network, regulate the fusion process. The efficiency of fusion is strikingly dependent on the composition of the isotonic medium, being relatively high at an orthophosphate concentration of 5 mM and minimal at 20 mM. PMID- 9394296 TI - Patenting inventions in the field of biology and chemistry: German and European patent law and case law. AB - Patent law is intended to provide protection for new and inventive achievements in technology. Technical progress is considered to be the purpose and aim of patent law. The main objective of patent law is to protect patentable results according to the latest state of science and research. It is most important to encourage the inventor to completely publish his knowledge. As a reward for this the inventor is granted a right of exclusion which is limited in time: the patent. It is not the purpose of patent law to enrich mere theory but to create industrially applicable knowledge for the public. This contribution deals with important problems to be considered by inventors in the patenting of inventions in the field of biology and chemistry. Such questions are related particularly to the accessibility of inventions and discoveries to patent protection, the various kinds (categories) of patents, the requirement of novelty, complete disclosure of the invention, patentability of DNA sequences and proteins, as well as inventive step. PMID- 9394297 TI - How to derive steady-state rate laws for enzyme-catalyzed reactions from macroscopic probabilities. PMID- 9394298 TI - Molecular intervention with antisense oligodeoxynucleotides (ODNs) in nephrology. PMID- 9394299 TI - How does the macula densa sense tubule function? PMID- 9394300 TI - Genetics of cardiovascular disease in type 1 (insulin-dependent) diabetes with and without renal involvement. PMID- 9394301 TI - Implications of the Systolic Hypertension in Europe (Syst-Eur) Trial for clinical practice. PMID- 9394302 TI - PTH--one can teach an old hormone new tricks. PMID- 9394303 TI - Malnutrition is bad, but how can one detect malnutrition? PMID- 9394304 TI - Apoptosis: background and possible role in secondary hyperparathyroidism. PMID- 9394305 TI - Nephrology, dialysis and transplantation in Brazil. AB - The European Renal Association welcomes this opportunity and feels that European nephrologists should be informed about the state of nephrology in South America, with which particularly our Latin colleagues maintain close cultural relationships. The following report on Nephrology in Brazil is a welcome addition to a series of reports designed to provide to European nephrologists a global view of nephrology. It is hoped that this is not misconstrued as a violation of nephrological Monroe doctrine. PMID- 9394306 TI - Preservation of renal function: the spectrum of effects by calcium-channel blockers. AB - The vast majority of animal data derived from models of either remnant kidney or diabetes demonstrate that dihydropyridine (nifedipine-like) calcium-channel blockers (DHPCCBs) effectively reduce arterial pressure but do not significantly affect proteinuria nor prevent development of glomerular scarring. Conversely, the non-DHPCCBs such as diltiazem and verapamil blunt both the rise in proteinuria as well as mesangial matrix expansion and subsequent glomerular scarring in diabetes. Additionally, the non-DHPCCBs markedly attenuate development of glomerular scarring in the remnant kidney model. The primary reasons for these differences between subclasses of CCBs relates to a lack of the following attributes by DHPCCBs: (1) they fail to reduce glomerular membrane permeability which is increased in these models; (2) they fail to affect the synthesis of certain key matrix proteins that perpetuate development of glomerular scarring (this effect may be due to the differential expression of calcium channels within the glomerular mesangium); and (3) the DHPCCBs totally abolish renal autoregulation in these models, an effect not observed with non DHPCCBs. Taken together with long-term (> 3 year) clinical studies, primarily in diabetic nephropathy, it is clear that the non-DHPCCBs seem to offer protection to the kidney not available with DHPCCBs alone, unless systolic arterial pressure is reduced to levels of < or = 110 mmHg. PMID- 9394307 TI - Organ donation in the UK: a survey by a British Transplantation Society working party. AB - BACKGROUND: During the past few years the number of organ donors in the UK has declined after a slow but steady increase during the 1980s. Concern about the decline led to a survey by the British Transplantation Society. The report of this survey highlighted a number of reasons for the decline and this manuscript presents and discuss the main items in the report. METHODS: Comprehensive information relating to organ donation was obtained by a combination of structured interviews during visits to intensive care units (ICUs) and neurosurgical units, the use of detailed questionnaires sent to all UK ICUs, and from the register held by the United Kingdom Transplant Support Service Authority. RESULTS: The information obtained highlighted a number of reasons for the decline in organ donor numbers and these are presented and discussed. The pool of potential donors is shrinking as death rates from road traffic accidents and intracranial haemorrhage decrease. Also the increasing use of modern imaging techniques has improved predictive ability in patients with severe brain damage with the result that more patients whose prognosis is assessed as hopeless are not treated by ventilation. Inadequacies both in intensive care unit bed provision and the resourcing of the transplant co-ordinator service were also thought to be important. CONCLUSIONS: Eight recommendations have been made, covering ICU bed provision, neurosurgical provision, transplant surgical staffing, the transplant co-ordinator network, reimbursement to donor units, asystolic donation, live donor transplantation, and interventional ventilation. PMID- 9394308 TI - Intercellular adhesion molecule-1 mediated interactions and leucocyte infiltration in IgA nephropathy. AB - BACKGROUND: Mononuclear leucocytes have a role in IgA nephropathy (IgAN). Renal leucocyte recruitment is mediated by adhesive interactions between leucocytes and their ligands on renal cells. METHODS: We have assessed interstitial and glomerular leucocytes by avidin-biotin-peroxidase with monoclonal antibodies (MA) against leucocytes (CD45), beta 2-integrin (CD18), monocyte-macrophages (CD14), T (CD3) and T-cell subsets (CD4, CD8), and intercellular adhesion molecule-1 (ICAM 1) (CD54), and analysed their relation with the abnormal expression of ICAM-1 on proximal tubule epithelium in sequential renal sections from 48 patients with IgAN stratified according to the severity of the interstitial cellular infiltration observed by light microscopy. RESULTS: In IgAN without (n = 15) and with (n = 7) interstitial cellular infiltration of 1+, ICAM-1 expression on vascular endothelium was unchanged with respect to that observed in the normal kidney; the proximal tubule epithelium was negative for this stain. In IgAN with interstitial cellular infiltration of 2+ (n = 10), 3+ (n = 11), and 4+ (n = 5), ICAM-1+ stain was observed on the proximal tubule epithelium, the median value of its quantitative expression being 0.3, 0.1, and 0.2 (P = 0.0008), respectively. The tubular ICAM-1 + stain was significantly associated with the interstitial leucocytes identified by MA, and correlated with CD45+ (r = 0.59, P = 0.02), CD14+ (r = 0.54, P < 0.02), and CD3+ (r = 0.51, P = 0.02) interstitial leucocytes in IgAN with interstitial cellular infiltration. Interstitial ICAM-1+ and CD18+ leucocytes were correlated (r = 0.56, P < 0.001). Correlation was found between the quantitative tubular expression of ICAM-1+ and the number of CD45+ (r = 0.98, P < 0.0001), CD3+ (r = 0.48, P = 0.02), and CD8+ (r = 0.76, P < 0.02) glomerular leucocytes. CONCLUSIONS: Our results suggest that tubular and interstitial ICAM 1+ cells may participate in adhesive interactions with interstitial leucocytes. Interstitial T-cells and macrophages as well as glomerular T-cells bearing predominantly CD8+ phenotype could play a role in the induction of the tubular expression of ICAM-1 in IgAN. PMID- 9394309 TI - Occurrence of anti-C1q antibodies in IgA nephropathy. AB - BACKGROUND: The pathogenic mechanisms and the antigens involved in the establishment and progress of IgA nephropathy are unknown. As antibodies against C1q have been reported to correlate with SLE nephritis, we analysed the occurrence of these antibodies in IgA nephropathy in order to investigate the possibility of pathogenetic similarities in these renal disorders. METHODS: The occurrence of IgA- and IgG anti-C1q antibodies (anti-C1q) were determined by ELISA in patients with IgA nephropathy (n = 36) and SLE nephritis (n = 37), diseases both known to be associated with circulating immune complexes. Levels of these antibodies were also determined in two other glomerular diseases, i.e. idiopathic membranous glomerulonephritis (n = 7) and minimal change disease (n = 2), in which circulating immune complexes are usually not present, and in 40 healthy controls. RESULTS: IgA anti-C1q was observed in increased titres in 11/36 of the patients with IgA nephropathy, in 2/37 of the patients with SLE nephritis (both with proliferative disease) and in 1/9 of the patients with membranous and minimal change disease (P < 0.001). Increased titres of IgG anti-C1q were observed in 1/36 of the patients with IgA nephropathy, in 17/37 of the patients with SLE nephritis and in 0/9 of the patients with membranous and minimal change disease (P < 0.001). There were no correlations between the levels of anti-C1q antibodies and clinical parameters such as degree of proteinuria, haematuria, or renal function. Nor was there any correlation to the concentration of C3a and the terminal complement complex (TCC) in patients with IgA nephropathy. CONCLUSIONS: The occurrence of anti-C1q antibodies in both IgA nephropathy and SLE nephritis, albeit of different predominating isotypes, indicates the possibility of a similar pathogenic mechanism involved in these renal disorders. The occurrence of IgA anti-C1q antibodies in patients with IgA nephropathy has to our knowledge not previously been reported. PMID- 9394310 TI - Properties of circulating IgA molecules in Henoch-Schonlein purpura nephritis with focus on neutrophil cytoplasmic antigen IgA binding (IgA-ANCA): new insight into a debated issue. Italian Group of Renal Immunopathology Collaborative Study on Henoch-Schonlein purpura in adults and in children. AB - BACKGROUND: The presence and the pathogenetic role of circulating IgA reacting with neutrophil cytoplasmic antigens (IgA-ANCA) in patients with Henoch-Schonlein purpura (HSP) is still debated. This study was aimed to investigate some characteristics of serum IgA and macromolecular IgA in HSP patients, focusing on IgA-ANCA. METHODS: Eighty-seven HSP patients with biopsy proved renal involvement (51 adults and 36 children) enrolled in a multicentre study of the Italian Group of Immunopathology were investigated. RESULTS: Significantly high levels of IgA immune complexes were found in both adults (P < 0.05) and children (P < 0.01), while the binding of IgA to jacalin, was significantly low in children with HSP (P < 0.01) only. Two series of ELISA were done for IgA-ANCA, in two different laboratories. Increased binding to PMN crude extracts (P < 0.01) without any modification in IgA binding to proteinase 3 was found by either specific ELISA. Conversely, the binding of IgA to myeloperoxidase (MPO) was found to be significantly (P < 0.05) increased with positive values in 25% of patients by one assay only. Three of four sera with positive IgA-MPO ANCA exhibited binding in Western-blot studies with the MPO preparation used in ELISA to a 28-kDa species. D-galactose and N-acetyl-glucosamine decreased the binding of serum IgA to MPO more in HSP than in controls (P < 0.05). CONCLUSIONS: The conflicting reports on IgA-ANCA may reflect some atypical characteristics of the reaction which can be detected only by some ELISAs. We suggest that not an antigen-antibody reaction but a lectinic interaction due to abnormal composition of IgA carbohydrate side chains may account for the IgA-ANCA reaction in patients with HSP nephritis. PMID- 9394311 TI - Long-term prognosis of Henoch-Schonlein nephritis in adults and children. Italian Group of Renal Immunopathology Collaborative Study on Henoch-Schonlein purpura. AB - BACKGROUND: The aim of this multicentre collaborative study was to compare the progression of renal disease in children and adults with Henoch-Schonlein purpura (HPS) nephritis selected on the basis of IgA-dominant renal deposits and biopsy material available for review. METHODS: The analysis was performed in 152 patients (95 adults and 57 children < 16 years old at diagnosis) with a follow-up (> or = 1 year up to 20 years (4.9 +/- 3.4 years in adults and 4.8 +/- 3.9 years in children). RESULTS: Renal histology and clinical presentation were similar in both age groups: crescents were found in 36% of adults and 34.6% of children (in only 2.7% of adults and 1.9% of children involving > 50% of glomeruli), nephrotic range proteinuria in 29.5% of adults and 28.1% of children and functional impairment in 24.1% of adults and 36.9% of children. The outcome was similar for both age groups (remission, 32.5% of adults and 31.6% of children; renal function impairment, 31.6% of adults and 24.5% of children). Endstage renal disease was observed in 15.8% of adults and in 7% of children. Renal function survival at 5 years was not significantly different in the two groups (85% in adults and 95% in children) and at 10 years it was approximately 75% in both groups. None of the children died and adult survival was 97% at 5 years. In adults at presentation, renal function impairment (P < 0.02) as well as proteinuria higher than 1.5 g/day (P < 0.02) and hypertension (P < 0.001) were negative prognostic factors. Multivariate analysis stressed the main statistical relevance of proteinuria (relative risk 2.37, P < 0.02). Conversely, in children no definite level of proteinuria, hypertension or other data were found to be associated with poor prognosis. CONCLUSIONS: Among patients with a clinical presentation which warrants renal biopsy, HSP nephritis has a similar prognosis in children and adults. The evolution is more predictable in adults than in children. PMID- 9394312 TI - Neonatal presentation of autosomal dominant polycystic kidney disease with a maternal history of tuberous sclerosis. AB - BACKGROUND: Childhood presentation of polycystic kidney disease has been reported with tuberous sclerosis complex (TSC). Recently some such cases have been shown to be due to combined deletion of the PKD1 and TSC2 genes, which lie close together on chromosome 16. The phenomenon of anticipation, whereby disease presentation occurs at a progressively earlier age in each generation, has been suggested to occur in autosomal dominant polycystic kidney disease (ADPKD). We have carried out a genetic study of a family in which these issues became clinically relevant. Neonatal presentation of polycystic kidneys occurred in an individual with a maternal family history of epilepsy and features of TSC without renal cystic disease. METHODS: Detailed historical and clinical profiles were gathered for three generations of the maternal and paternal families. Both parents underwent renal ultrasound scanning. Genomic DNA was obtained from affected and unaffected individuals from the maternal family and used for linkage analysis to gene loci for TSC. RESULTS: Renal cysts were not present in the mother by ultrasound. Linkage to TSC2 was found for members of the maternal family with clinical features of TSC. While a diagnosis of TSC was confirmed in her mother the child was found not to have inherited the disease-related allele. The father was found to have asymptomatic bilateral polycystic kidneys consistent with ADPKD. The presence of ADPKD in other paternal relatives could not be confirmed. CONCLUSIONS: The index case was found to have paternally inherited ADPKD with unusually early presentation. While at risk for concomitant maternal inheritance of TSC this diagnosis was ruled out by linkage analysis studies. The ability to clarify the true nature of a complex inherited condition greatly facilitates future management and counselling. The mechanisms underlying phenotypic heterogeneity in ADPKD remain to be clearly defined and are the subject of ongoing investigation. PMID- 9394313 TI - Enalapril versus metoprolol in primary hypertension--effects on the glomerular filtration rate. AB - BACKGROUND: Hypertension is a significant cause of end-stage renal failure and effective treatment of hypertensive will reduce the progression rate of chronic renal failure in various kidney disorders. Different classes of drugs may be more effective than others in this respect. In this study we compared the effects on the glomerular filtration rate (GFR) of the ACE-inhibitor enalapril and the betablocker metoprolol in patients with mild and moderate primary hypertension during 6 years. METHODS: Patients with GFR in the normal range (> or = 80 ml/min/1.73 m2 BSA) were included after a placebo treatment period of 4-8 weeks if diastolic blood pressure was 100-120 mm Hg. Target blood pressure was set to < 90 mm Hg diastolic. One hundred and thirty patients were randomized in an open parallel study to receive either enalapril or metoprolol. No placebo group was included. GFR was measured using the 51CR-EDTA clearance method and 81 patients completed the study. RESULTS: At inclusion, there were no significant differences regarding GFR or blood pressure between the groups. The blood pressure treatment goal was reached in all patients and was maintained during the whole observation period. A small but significant fall in GFR by 4 ml/min/1.73 m2 BSA was noted in both groups after the first year of treatment but thereafter GFR decreased by only 1 ml/min/year/1.73 m2 BSA, in both groups. Body weight, serum uric acid and triglycerides increased slightly with metoprolol treatment but no other differences between the two treatments were noted. CONCLUSIONS: With the blood pressure maintained at the same level using either enalapril or metoprolol during a 6-year study period, GFR decreased to the same extent in the two groups both during the first year and thereafter. The overall magnitude of the GFR decline approached that of the normal age-related decrease of kidney function, i.e. GFR decreased only about 1 ml/min/year. Thus, treatment with an ACE-inhibitor, enalapril, and a beta-blocker, metoprolol, protected the kidney function to the same extent in this 6 year long study in mild and moderate primary hypertension. PMID- 9394314 TI - A randomized, controlled study of sulodexide therapy for the treatment of diabetic nephropathy. AB - BACKGROUND: Glycosaminoglycans (GAGs) play an important role in the physiopathology of diabetic nephropathy; they are essential for the maintenance of glomerular charge selectivity and their administration can reduce albuminuria in diabetic patients. METHODS: Following a randomized block design, controlled versus placebo, we investigated, in insulin-dependent diabetic patients with micro- or macroalbuminuria, whether GAG therapy can influence an altered albumin excretion rate (AER). Thirty-six patients (18 micro- and 18 macroalbuminuric) were randomized to receive, during 5 days/week for 3 weeks, either a daily dose of 600 lipoproteinlipase releasing units (LRU) of sulodexide by the i.m. route (9 micro- and 9 macroalbuminuric patients), or a matching i.m. placebo (9 micro- and 9 macroalbuminuric patients). All patients were followed-up for further 6 weeks. AER was evaluated before treatment, weekly during it and every 3 weeks during follow-up. RESULTS: Seventeen of the 18 sulodexide-treated patients showed a trend towards decrease in AER, more evident and statistically significant in microalbuminurics (P < 0.01 after the first week). At the end of follow-up, AER was still significantly reduced in microalbuminurics, while macroalbuminurics showed again increased values. Placebo-treated patients evidenced no AER variations during all the study period. No statistically significant differences vs baseline, concerning blood pressure, haematological, haematochemical, and coagulative tests, and urinalysis, were ever observed, apart from a clear-cut decrease in blood cholesterol and triglycerides at the end of treatment, in a subgroup of hyperlipidaemic, sulodexide-treated subjects. No adverse events were registered. CONCLUSIONS: Our results suggest that the GAG sulodexide exerts a positive activity in type I diabetic patients with micro- and macroalbuminuria, by reducing the abnormally high AER levels. PMID- 9394315 TI - An investigation of the effect of advancing uraemia, renal replacement therapy and renal transplantation on blood pressure diurnal variability. AB - BACKGROUND: Ambulatory blood pressure recordings have been shown to correlate better with target organ damage than have isolated clinic blood pressure readings. There have been some small studies demonstrating that abnormal blood pressure diurnal rhythm is common in uraemia and in patients on renal replacement therapy. Abnormal blood pressure diurnal rhythm itself may be a risk factor for accelerated target organ damage. METHODS: We retrospectively studied 480 ambulatory blood pressure recordings in 380 patients with essential hypertension, secondary hypertension, and on renal replacement therapy. We examined diurnal blood pressure rhythm in each group. RESULTS: Abnormal blood pressure diurnal rhythm (non-dipping) is significantly more prevalent in patients with underlying renal disease, even with normal excretory renal function (53%) than in age-, sex , and race-matched controls with essential hypertension ((30%), P < 0.01). In patients with renal disease the prevalence of non-dipping rose with worsening renal function, reaching statistical significance once plasma creatinine was greater than 400 mumol/l. There was a direct correlation between plasma creatinine and percent decline in blood pressure at night for both systolic (r = 0.23) and diastolic (r = 0.24) blood pressure in patients with underlying renal disease and impaired excretory renal function. High prevalences of abnormal diurnal BP rhythm are seen in patients on haemodialysis (82%), peritoneal dialysis (78%), patients with plasma creatinine > 600 mumol/l (75%), and in renal transplant recipients (74%). CONCLUSIONS: Abnormal blood pressure diurnal rhythm ('non-dipping') is significantly more common in secondary than in primary hypertension, even with normal renal function. Abnormal blood pressure diurnal rhythm becomes increasingly common with advancing uraemia. Once the plasma creatinine is greater than 600 mumol/l the prevalence of non-dipping is the same as that seen with renal replacement therapy. This phenomenon is not modulated by successful renal transplantation. PMID- 9394316 TI - Interdialytic weight gain and 48-h blood pressure in haemodialysis patients. AB - BACKGROUND: Hypertension, which is often associated with hypervolaemia, is common in haemodialysis patients and is a known determinant of target organ damage. Interdialytic weight gain due to volume overload has also been associated with mortality in haemodialysis patients. METHODS: We therefore studied 27 chronic haemodialysis patients who underwent 48-h ambulatory blood pressure monitoring between two midweek dialysis sessions, and 2D and M-mode echocardiography for determination of left ventricular mass index. RESULTS: Left ventricular hypertrophy (left ventricular mass index in men > 131 g/m2, women > 100 g/m2) was present in 70% (19/27) patients despite a mean 48-h blood pressure of 132 +/- 19/81 +/- 15 mmHg. Mean interdialytic weight gain was 1.6 +/- 0.8 kg and was not related to left ventricular mass index. Two patterns of interdialytic blood pressure change were apparent: in group 1 (16 patients) 48-h blood pressure increased (+19 +/- 12/13 +/- 9 mmHg), whereas in group 2 (11 patients) blood pressure fell (-10 +/- 13/-8 +/- 10 mmHg P < 0.0001). In both groups the number of hypertensive patients (group 1, 10/16; group 2, 6/11), the 48-h blood pressure (132 +/- 20/80 +/- 15 vs 132 +/- 18/82 +/- 15 mmHg) and interdialytic weight gain (+1.9 +/- 0.7 vs +1.3 +/- 0.7 kg) were similar. There was also no correlation between interdialytic blood pressure change and weight gain in either group. CONCLUSIONS: We conclude that interdialytic blood pressure changes cannot be directly related to interdialytic fluid gain, even in apparent volume-dependent hypertension, emphasizing the importance of additional factors in the control of blood pressure in end-stage renal disease. PMID- 9394317 TI - Erythropoietin and oxidative stress in haemodialysis: beneficial effects of vitamin E supplementation. AB - Oxidative stress can produce profound alterations to cellular membrane lipids, impairing cell metabolism and viability. This phenomenon, previously observed in haemodialysis patients, has been proposed as a significant factor in regard to haemodialysis-related shortened red blood cells (RBC) survival. In the present study, several parameters associated with oxidative stress were evaluated in a group of haemodialysis patients either receiving erythropoietin therapy (n = 12, mean erythropoietin dose 88 +/- 24 U/kg/week) or not receiving such therapy (n = 30), and in 38 controls. Malonyldialdehyde (MDA, nmol/ml), an end-product of lipid peroxidation, and RBC antioxidant systems were measured, including RBC alpha-tocopherol (RBC vitamin E, mg/l), RBC glutathione (GSH, nmol/mgHb), and RBC superoxide dismutase activity (SOD, U/mgHb). Plasma vitamin E concentrations were also evaluated. Finally, oral vitamin E supplementation (500 mg daily), an exogenous antioxidant, was administered for 6 months to seven patients from the dialysis group receiving erythropoietin while oxidative parameters were repeatedly evaluated and erythropoietin requirements monitored, in order to appreciate the therapeutic relevance of an antioxidant supplementation. An elevation of serum MDA was observed in all haemodialysis patients and a significant decrease in RBC vitamin E, despite normal serum vitamin E levels. Furthermore, the reduction in RBC vitamin E was more important in patients treated with erythropoietin. Vitamin E supplementation resulted in a significant increase in RBC vitamin E (from 0.3 +/- 0.1 to 1.2 +/- 0.2 mg/l of pellet) and a reduction in erythropoietin dose (from 93 +/- 24 to 74 +/- 26 U/kg/week) while maintaining stable haemoglobin concentrations. These results suggest that the oxidative stress could be one of the resistance factors to erythropoietin response in haemodialysis and that vitamin E supplementation could have a sparing effect on erythropoietin dosage requirement. PMID- 9394318 TI - Increased plasma leptin/fat ratio in patients with chronic renal failure: a cause of malnutrition? AB - BACKGROUND: Protein-energy malnutrition occurs in patients with chronic renal failure primarily due to loss of appetite. The ob gene protein, leptin, which is secreted by adipocytes, regulates body composition by lowering food intake. We have measured plasma leptin in undialysed and dialysed patients and in controls and the concentrations have been related to body composition, dietary intake, and biochemistry. METHODS: Plasma leptin was measured by radioimmunoassay in 93 individuals in groups of undialysed, peritoneal dialysed, and haemodialysed patients and controls. Body composition was determined by DEXA. RESULTS: Protein energy malnutrition was evident in non-dialysed and dialysed patients from low lean or fat tissues, plasma albumin and transferrin. A third of the dialysis patients were eating less than prescribed intakes. Leptin relative to total fat mass (ng/ml/kg) was significantly greater for patients than for controls, particularly the dialysed patients. Leptin was highly correlated with total, arm, leg, and all other fat measurements, e.g. r for leptin vsm % total fat was: undialysed 0.88, PD 0.81, HD 0.93, and controls 0.83 (P < 0.0001 for all). Dialysis patients with the highest leptin/fat mass ratio had low protein intakes and significantly lower lean tissue mass. Leptin/fat ratio correlated inversely with dietary intake e.g. with protein intake in g/day and marginally in g/kg of ideal weight/day. Leptin concentration was unrelated to plasma creatinine or residual renal function or to the protein 'nutritional indices', albumin and transferrin. CONCLUSIONS: Our data suggests that leptin is markedly increased in some patients with chronic renal failure. The association of increased leptin with low protein intake and loss of lean tissue is consistent with leptin contributing to malnutrition but a definitive role cannot be substantiated by this study. PMID- 9394319 TI - The impact of malnutrition in morbidity and mortality in stable haemodialysis patients. Spanish Cooperative Study of Nutrition in Hemodialysis. AB - BACKGROUND: When assessed by single biochemical measurements, malnutrition in dialysis patients is associated with increased mortality, but there are few data evaluating abnormalities in anthropometry or composite nutritional scores and outcome. The aim of our study was to ascertain the prevalence and severity of malnutrition in 761 stable patients from 20 haemodialysis centres and its influence in morbidity and mortality after one year of follow-up. METHODS: Malnutrition was estimated by scoring four anthropometric indexes; body mass index (BMI), triceps skinfold thickness (TSF), mid-arm circumference (MAC), and mid-arm muscle circumference (MAMC); three biochemical measurements; serum albumin, serum transferrin and total lymphocyte count; and clinical examination. Mortality and hospitalizations were collected prospectively during the year of follow-up. RESULTS: A moderate/severe degree of malnutrition was presented by 51.6% of male and 46.3% of female patients. TSF was moderate-severely decreased in 41% without differences between males and females. MAMC was moderately decreased in 19.8% of males and in 8.1% of females (P < 0.001). Multiple logistic regression analysis showed that the predictors of malnutrition were: age > 65 years (OR = 1.96, CI: 1.22-3.14), male sex (OR = 1.95, CI: 1.24-3.07), comorbidity index (OR = 1.23, CI: 1.03-1.45), time on dialysis (OR = 1.13, CI: 1.08-1.18), duration of dialysis (OR = 0.73, CI: 0.63-0.85) and PCR related to ideal body weight (OR = 0.17, CI: 0.06-0.50). After 1 year of follow-up, data from 442 patients were available. A total of 68 patients died (15.4%) with cardiovascular diseases being the most frequent cause of death (57.3% of the cases). The predictors of mortality were: age (OR = 1.06, CI: 1.03-1.09), cardiovascular disease (OR = 2.13, CI: 1.19-3.83), neurological disease (OR = 2.96, CI: 1.41-6.15), nephroangiosclerosis (OR = 2.34, CI: 1.10-4.98) and total lymphocyte count (OR = 0.93, CI: 0.87-0.98). Hospitalization was needed in 44% of patients. The comorbidity index, serum albumin and age were the predictive factors of hospitalization. CONCLUSIONS: Protein-calorie malnutrition is frequent in haemodialysis patients. Fat depletion predominated in both sexes. Duration of dialysis and protein catabolic rate related to ideal body weight was the only predictor which could be influenced by a nutritional intervention. Morbidity appeared to be influenced by the comorbidity index and age was the strongest predictor of mortality. The only nutritional measurements predictive of morbidity and mortality were serum albumin and total lymphocyte count respectively. Therefore, the influence of malnutrition in morbidity and mortality can not be definitively stated. PMID- 9394320 TI - Total, free, and protein-bound sulphur amino acids in uraemic patients. AB - Fasting plasma concentrations of sulphur amino acids (sAA) were measured in nine non-dialysed (ND) chronic uraemic patients on conservative treatment, 10 patients on continuous ambulatory peritoneal dialysis (CAPD), nine patients on haemodialysis (HD) treatment, and 10 healthy subjects (HS). Methionine and taurine concentrations were significantly decreased in the CAPD and HD patients and tended to be low in the ND patients. Cysteine sulphinic acid (CSA) levels were significantly higher in all patient groups. Total (t), free (f), and protein bound (pb) homocysteine (Hcy) and cysteine (Cys) were significantly increased in all patient groups. Serine, a substrate for cystathionine synthesis from Hcy, showed significantly lower concentrations in all patient groups. The percentages of pbHcy were significantly higher in the CAPD and HD patients than in the ND patients (P < 0.0001, P = 0.002 respectively) or in the HS (P < 0.0001, P = 0.008 respectively), whereas the percentages of pbCys in CAPD and HD patients were significantly higher than in ND patients (P = 0.0006, P = 0.009 respectively) and tended to be high without reaching statistical significance compared to the HS. A single HD treatment decreased tHcy by 26%, fHcy by 39%, and pbHcy by 22%, as well as tCys by 40%, fCys by 54%, and pbCys by 27%. The tHcy concentration, although decreased by HD treatment, remained higher than in HS, whereas tCys was normalized by the dialysis session. In addition, HD treatment significantly decreased the plasma concentrations of methionine, CSA, taurine, and serine. We conclude that, except for methionine and taurine, the plasma sAA in their different forms are markedly increased in dialysed and non-dialysed uraemic patients. The percentages of pbHcy and pbCys were significantly higher in dialysed than in ND uraemic patients. HD treatment can normalize the tCys concentration, and decrease the tHcy concentration but not normalize it. The observed hyperhomocysteineaemia and low taurine levels may contribute to the high incidence of cardiovascular disease in uraemic patients. PMID- 9394321 TI - Free amino-acid levels simultaneously collected in plasma, muscle, and erythrocytes of uraemic patients. AB - BACKGROUND: Disturbances in amino acid (AA) metabolism in uraemia have mainly been reported to occur in plasma and muscle. The erythrocytes (RBC) constitute a large proportion of the free AA in blood and may play an important role in the interogan transport of AA. This report presents the first data on AA levels obtained simultaneously from three different compartments in uraemic patients. METHODS: Muscle biopsy and blood samples were obtained from 38 haemodialysis (HD), 22 continuous peritoneal dialysis (CPD) and 10 end-stage renal failure patients for determination of free amino acids by reversed-phase HPLC. The results are compared to data obtained from 27 healthy subjects under the same conditions. RESULTS: For a number of non-essential AA (alanine, glycine, asparagine, arginine) and for lysine, elevated concentrations were present simultaneously in RBC and in muscle but not in plasma. On the other hand, low concentration of some essential AA (leucine, valine, phenylalanine, tyrosine) were observed in RBC and in plasma, while the concentrations in muscle were normal. Most of the non-essential AA (NEAA), especially taurine and glutamine, had much higher muscle/plasma gradients than RBC/plasma gradients, although an accumulation in RBC of glycine, serine, arginine, asparagine, ornithine, glutamate and taurine was observed. Most of the essential AA (EAA) showed higher muscle/plasma gradients, whereas the RBC/ plasma gradients were approximately 1.0. CONCLUSION: Our findings are in agreement with studies that have shown that RBC and plasma play independent and opposing roles in AA interorgan transport. The results indicate that there are several AA abnormalities in all three compartments in uraemic patients. They also suggest that there may be some specific common changes of selected transport systems for both RBC and muscle in uraemia. Determination of AA in RBC should be considered when undertaking metabolic and clinical studies of AA disturbances. PMID- 9394322 TI - Individualized anticoagulation with dermatan sulphate for haemodialysis in chronic renal failure. AB - BACKGROUND: Dermatan sulphate (DS) is a selective thrombin inhibitor with antithrombotic properties and low bleeding potential. In preliminary studies it was reported to be effective for preventing clot formation in the haemodialysis circuit. METHODS: Ten patients on maintenance haemodialysis for chronic renal failure underwent three consecutive investigation phases. In phase 1 (individual dose titration), repeated dialyses were performed with increasing doses of DS until successful dialysis was obtained in two sessions at the same dose. In phase 2, individualized DS doses were validated by a randomized crossover comparison with the individual heparin dose of each patient. In phase 3, each patient underwent 24 consecutive dialyses with DS over 8 weeks. Successful dialysis was defined as completion of the procedure without visible clot formation in the bubble traps and lines or a greater than 20% decrease in dialyser capacity. Dialysis efficiency (decrease in serum urea and creatinine, Kt/V), APTT prolongation, bleeding time, and DS plasma concentrations were also assessed. RESULTS: Phase 1: successful dialysis was achieved in nine patients with 4 mg/kg DS as a predialysis intravenous bolus followed by continuous infusion of 0.65 mg/kg/h. One patient required 5 mg/kg plus 1.3 mg/kg/h. Phase 2: no statistically significant differences were found between DS and heparin in any of the investigated variables. Residual dialyser capacity and dialysis efficiency indexes indicated equivalent efficacy. Phase 3: residual dialyser capacity and dialysis efficiency did not change with time. There was no accumulation of DS in plasma. No bleeding or thrombocytopenia were observed. CONCLUSIONS: The dose of DS can be individually titrated to suppress clot formation during haemodialysis as efficiently as with individualized heparin. Such an individualized DS regimen maintains its anticoagulant efficacy and is safe in prolonged use. PMID- 9394323 TI - Plasma hypercoagulability in haemodialysis patients: impact of dialysis and anticoagulation. AB - BACKGROUND: Thrombotic complications are common in patients with endstage renal disease and contribute substantially to the morbidity and mortality in this population. The aim of the present study was to: i) determine the prevalence and the extent of hypercoagulability in patients undergoing dialysis treatment by measuring parameters that directly reflect thrombin concentrations; ii) assess changes in coagulation status during haemodialysis (HD); iii) quantify the relative impact of heparin, dialysis and their combined effects on coagulation status and iv) detect factors that modify coagulation haemostasis in dialysis patients. METHODS: A total of 39 patients (HD: n = 29, CAPD: n = 10) was analysed for procoagulatory and fibrinolytic activity determined by measurements of partial thromboplastin time, prothrombin fragments F1 + 2, thrombin-antithrombin complexes and D-dimer concentrations. HD patients were investigated prior to and during dialysis. A subgroup of patients was infused heparin alone without dialysis or was dialysed without heparin administration. Furthermore, subgroup and correlation analyses were performed for the type of dialysis (HD vs CAPD), dialyzer and shunt, Kt/V, underlying disease and treatment with recombinant erythropoietin (rhEPO). RESULTS: Baseline levels of all parameters-procoagulatory and fibrinolytic--were substantially elevated in all patients, but to a higher degree among those on CAPD. Moreover, haemodialysis treatment increased procoagulatory markers even further, suggesting stimulated coagulation and/or insufficient anticoagulation during dialysis. However, after 3 h of dialysis thrombin concentrations, determined by quantification of prothrombin fragments, were inversely correlated with Kt/V. Selective heparin infusion diminished procoagulatory activity only slightly and incompletely, whereas HD without heparin resulted in excess thrombin accumulation. Finally, subgroup analyses revealed more pronounced thrombin formation among patients treated with polysulfon dialyzers, whereas erythropoietin dosage was positively related with lower procoagulatory activity. CONCLUSION: A majority of patients on dialysis are in a hypercoagulable state, which is further aggravated by the haemodialysis procedure itself and may not be sufficiently controlled with current anticoagulation regimens. Intensified heparin treatment and the use of rhEPO are likely to improve coagulation haemostasis, whereas the type of dialyzer should be considered as a relevant procoagulatory factor. PMID- 9394324 TI - Dissociation between beta-2 microglobulin and IL-1 production in hemodialyzed patients. AB - BACKGROUND: beta-2 microglobulin is predominant in amyloid deposits in patients undergoing long term hemodialysis. Amyloid accumulation has been ascribed to dialysis membranes, endotoxin contamination of the dialysate, uremia and chronic systemic inflammation associated with enhanced monocytic cytokine production in hemodialyzed patients. Interleukin-1 has been proposed to play a critical role in the induction of beta-2 microglobulin synthesis and release. METHODS: We examined if monocytes contribute to beta-2 microglobulin production upon stimulation with inflammatory mediators that are generated during hemodialysis and investigated the production of beta-2 microglobulin by cells from patients, with and without clinical signs of amyloidosis, at the time when patients' monocytes contained maximal intracellular accumulation of IL-1. RESULTS: We demonstrated that only monocytes are able to release increased levels of beta-2 microglobulin upon stimulation by IL-1, TNF alpha, C5a and LPS. Increased levels of beta-2 microglobulin were associated with increased levels of beta-2 microglobulin mRNA. Before dialysis session, 20-60% of circulating CD14+ monocytes from patients contained IL-1. At the time when maximal IL-1 production was detected, we showed by RT-PCR increased transcription of IL-1 gene in patients' monocytes. We observed that monocytes from patients with amyloidosis contained higher amounts of IL-1 as compared to monocytes from patients without clinical signs of amyloidosis, but could not secrete increased amounts of beta-2 microglobulin upon LPS-stimulation. CONCLUSIONS: Our data indicated that chronic inflammation, as demonstrated by increased intracellular IL-1 expression, is not associated with increased production of beta-2 microglobulin by monocytes from patients on hemodialysis. PMID- 9394325 TI - The required dose of erythropoietin during renal anaemia treatment is related to the degree of impairment in erythrocyte deformability. AB - BACKGROUND: Renal anaemia is rapidly corrected by recombinant human erythropoietin (rHuEpo) therapy, but the dose required varies greatly. Since impaired erythrocyte deformability may be one factor contributing to the development of renal anaemia, the interrelationship between that variable and the rHuEpo requirement was examined. METHODS: Twenty-five patients treated with hemodialysis and rHuEpo for at least 6 months were included in the study. The Hb value had been stable and the rHuEpo dose unchanged the last two months. Using a rotational viscometer, the fluidity of erythrocytes, separated from plasma and re suspended in isotonic buffered saline to a standardized haematocrit, was taken as a measure of erythrocyte deformability. RESULTS: The average weekly dose of s.c. epoetin alpha was 186 +/- 93 U/kg body weight (range 56-370). The dose was correlated to the reticulocyte fraction (R = 0.69, P = 0.0001). When the rHuEpo dose was used as dependent variable and blood haemoglobin concentration, serum (S) albumin, S ferritin, S aluminium, S PTH, S urea, Kt/V/week, erythrocyte fluidity, and plasma viscosity were used as independent variables in a stepwise multiple regression analysis, only erythrocyte fluidity remained significantly negatively correlated to the rHuEpo dose (R = 0.5, P = 0.01). Despite a tendency towards higher doses of rHuEpo in patients with a C-reactive protein concentration exceeding 20 mg/l, the Hb was lower in these patients. CONCLUSIONS: We conclude that the interindividual differences in bone marrow response to rHuEpo were small in these patients. Impaired erythrocyte deformability and inflammation seem to be factors associated with increased rHuEpo requirement. PMID- 9394326 TI - GBV-C/HGV infection in renal dialysis and transplant patients. AB - BACKGROUND: Recently a new human virus (GBV-C/HGV) was identified. With the use of the polymerase chain reaction (PCR) the possibility of a high prevalence of the GBV-C/HGV infection in haemodialysis patients was demonstrated. The aim of the present study was to use a combination of the PCR and a new diagnostic test for antibodies to the viral envelope protein E2 to assess the prevalence of the GBV-C/HGV infection in German patients with renal failure. METHODS: RT-PCR and ELISA were used to detect GBV-C/HGV RNA and antiviral antibodies (anti-E2) in the sera of 31 patients on a maintenance haemodialysis (HD)), 25 patients on a perioneal dyalisis (CAPD), and 92 renal transplant patients (RT). RESULTS: A statistical trend was noted for a higher rate of the GBV-C/HGV RNA in the whole group of patients with renal failure (11.5%) than in the control group of organ donors (5.5%); The difference between GBV-C/HGV RNA prevalence in RT patients (15.2%) and in organ donors (5.5%) was found to be significant. Anti-E2, which are considered as an indicator of a past GBV-C/HGV infection, were detected in 12.9% of HD patients, in 20.0% of CAPD patients, in 10.9% of RT patients, in 11.1% of organ donors, and in 10.9% of blood donors. These differences were not significant. Time on haemodialysis was significantly longer in GBV-C/HGV infected patients compared to uninfected patients and all patients with the GBV-C/HGV RNA have a history of blood transfusions. CONCLUSIONS: Patients with renal failure treated with dialysis or subjected to renal transplantation are at increased risk of aquiring the GBV-C/HGV infection. Higher rates of the GBV-C/HGV RNA and a similar prevalence of anti-E2 in patients with renal failure as compared to donors suggest that the rate of GBV-C/HGV infection resolution in immunosuppressed patients with renal failure might be lower than in immunocompetent patients. PMID- 9394327 TI - Rejection rates in kidney transplant patients with and without IgA nephropathy. AB - BACKGROUND: Based on graft survival rates it has been claimed that patients with IgA nephropathy have a reduced risk of rejection after kidney transplantation. We wanted to evaluate this hypothesis. METHODS: Certified IgA nephropathy was the original disease in 70 of 874 consecutive kidney transplant patients (8.0%). Eighty per cent of the patients were men. Median age was 37 years, range 9-64. Fifty-three per cent had living donors and 20% of the transplantations were pre emptive. Non-diabetic patients matched for age, sex, type of donor, and transplant number served as controls. Median follow-up time was 68 months. Duration of treatment for rejection during the first year post-transplant and graft loss due to rejection was recorded. RESULTS: The fraction of patients treated for rejection during the first year was 53% versus 54% of controls and the number of days when any antirejection treatment was given was 5.0 +/- 7.5 versus 5.5 +/- 7.4. Actual 3-year graft survival was 81% versus 80% and the number of grafts lost due to rejection was 9 versus 11. CONCLUSIONS: Rejection rates were not reduced in patients with IgA nephropathy and survival of grafts and patients not better than for matched controls. PMID- 9394328 TI - Colchicine myopathy in renal transplant recipients on cyclosporin. AB - Few data are available about the muscle status in renal transplant recipients. Moreover, the list of myotoxic drugs is growing longer and some of them are likely to be prescribed in renal transplant patients. These conditions may act as confounding factors in case reports of both cyclosporin and colchicine myopathies. Moreover no study has evaluated the frequency of myopathy in patients on both colchicine and cyclosporin. We conducted a retrospective study including all renal transplant recipients followed in our unit in whom colchicine was prescribed since January 1994. Clinical and biological data of patients on both colchicine and cyclosporin were analysed. Secondly case subjects were compared with matched controls not receiving colchicine but cyclosporin. Ten patients received colchicine in association with cyclosporin. Five patients (50%) experienced muscular symptoms and when performed, muscular histology showed vacuolar myopathy. All five patients improved after colchicine withdrawal. Cases with and without muscular symptoms did not differ in age, transplant duration, and serum creatinine level. Mean duration of colchicine therapy was 12.2 +/- 4.4 months in cases with muscular symptoms and 6.8 +/- 4.6 months in cases without muscular symptoms (P < 0.05). No control complained of either muscular pain nor weakness (P < 0.0005). Only one patient (3.3%) had elevated serum creatine phosphokinase concentration (P < 0.0005). We conclude that myopathy is very frequent in patient on both colchicine and cyclosporin. Muscular symptoms improve in all patients after colchicine withdrawal. PMID- 9394329 TI - Parathyroidectomy after renal transplantation: a retrospective analysis of long term outcome. AB - BACKGROUND: Advanced hyperparathyroidism refractory to active vitamin D continues to be a problem and frequently forces the nephrologist to resort to parathyroidectomy. One particular aspect is persisting advanced hyperparathyroidism after renal transplantation. Published information on this point is fragmentary. DESIGN: Retrospective analysis. PATIENTS: Between 1983 and 1995 a total of 456 patients with renal secondary hyperparathyroidism were subjected to parathyroidectomy (PTX) of whom 103 were transplanted or had at least a history of renal transplantation. The present analysis concerns 37 patients who had a functional renal graft at the time of PTX and were followed for up to 13 years. PTX was performed after an average of 36.7 months after renal transplantation. OUTCOME: Thirteen patients experienced rejection and became dialysis-dependent. Twenty-four patients had stable function of the renal graft. Seven patients died during follow-up. Hypoparathyroidism post-PTX developed in 4/37 patients, but could be overcome by replantation of cryoconserved parathyroid tissue. FREQUENCY ESTIMATE: A total of 2632 renal transplants were performed in the catchment area. As a minimum estimate 3.91% of patients with a functional graft required PTX. RECOMMENDATION: Parathyroidectomy should be considered early in cases with advanced secondary renal hyperparathyroidism, since renal transplantation does not necessarily guarantee reversibility of parathyroid overactivity. PMID- 9394330 TI - The influence of bicarbonate supplementation on plasma levels of branched-chain amino acids in haemodialysis patients with metabolic acidosis. AB - BACKGROUND: It has been hypothesized that correction of metabolic acidosis might improve the nutritional state of acidotic haemodialysis (HD) patients partly because of a reduced oxidation of branched-chain amino acids (BCAA). AIM: We investigated whether bicarbonate (Bic) supplementation in acidotic HD patients results in increased plasma levels of BCAA. METHODS: In a longitudinal study (run in period, 2 months; study period, 6 months), the effect of Bic supplementation on plasma levels of BCAA was studied in 12 acidotic HD patients (7 men, 5 women, mean age 54 +/- 18 years) with a predialysis bicarbonate (Bic) concentration smaller or equal to 22 mmol/l. Bic was supplemented by increasing Bic concentration of the dialysate and by oral Bic supplementation. RESULTS: Predialysis Bic increased significantly during the study period (18.7 +/- 2.7 vs. 23.1 +/- 11.5 mmol/l). There was no change in nutritional parameters. However, plasma levels of the BCAA valine, leucine, and isoleucine increased significantly. CONCLUSIONS: In haemodialysis patients with metabolic acidosis, Bic supplementation over a 6-months period resulted in an increase in plasma levels of BCAA. Further study is needed to elucidate the mechanisms behind, and the clinical importance of the observed changes in plasma BCAA levels. PMID- 9394331 TI - Cyclosporin A therapy in frequently relapsing nephrotic syndrome and IgA nephropathy. PMID- 9394332 TI - Membranoproliferative glomerulonephritis (MPGN) and pulmonary carcinoid tumour. PMID- 9394333 TI - Treatment of idiopathic retroperitoneal fibrosis with tamoxifen. PMID- 9394334 TI - Stretching of renal artery in a functionally solitary kidney: an unusual case of ischaemic nephropathy. PMID- 9394335 TI - Acute interstitial nephritis secondary to omeprazole. PMID- 9394336 TI - Functional renal recovery after spontaneous renal embolization in a sole kidney. PMID- 9394337 TI - Renal amyloidosis and renal failure--a novel complication of the SAPHO syndrome. PMID- 9394338 TI - Heparin-induced systemic inflammatory response syndrome with progressive skin necrosis in haemodialysis. PMID- 9394339 TI - Relapsing bacteraemia due to Micrococcus luteus in a haemodialysis patient with a Perm-Cath catheter. PMID- 9394340 TI - Recurrence of lecithin cholesterol acyltransferase deficiency after kidney transplantation. PMID- 9394341 TI - Tacrolimus treatment for steroid- and cyclosporin-resistant minimal-change nephrotic syndrome. PMID- 9394342 TI - Immune thrombocytopenic purpura presenting in an immunosuppressed patient after renal transplantation. PMID- 9394343 TI - Retropharyngeal abscess in a renal transplant recipient. PMID- 9394344 TI - Recurrence of lipoprotein glomerulopathy after renal transplantation. PMID- 9394345 TI - A transplanted child with severe hypercalcaemic hyperparathyroidism despite only modest bone lesions. PMID- 9394346 TI - Bacillus cereus peritonitis in a patient being treated with continuous ambulatory peritoneal dialysis. PMID- 9394347 TI - Doppler ultrasound in renal transplantation. PMID- 9394348 TI - A calcareous calamity. PMID- 9394349 TI - The alcoholic patient with an acute nephrotic syndrome and resistance to diuretics. PMID- 9394350 TI - Ultrasonographic evaluation of parathyroid hyperplasia. PMID- 9394351 TI - Sulphonamides in vasculitides: which mechanism? PMID- 9394352 TI - Calcifying panniculitis or 'simple' inflammation? Biopsy is better than a bone scan. PMID- 9394353 TI - Interleukin-6 production by endothelial cells: effect of corticosteroids. PMID- 9394354 TI - ANCA in Behcet's disease. PMID- 9394355 TI - Anti-beta 2-glycoprotein I and anti-prothrombin antibodies in haemodialysis patients. PMID- 9394356 TI - Hepatitis G virus infection in haemodialysis patients. PMID- 9394357 TI - Mycophenolate mofetil toxicity in an anorexic kidney transplant patient treated with sulphinpirazone. PMID- 9394358 TI - [Derivative advantages of autologous blood transfusion]. PMID- 9394359 TI - [Bone morphogenetic protein receptors]. PMID- 9394360 TI - [Free radicals and joint destruction]. PMID- 9394362 TI - A case of parathyroid carcinoma visualized on Tc-99m-sestamibi scintigraphy. AB - Recent studies indicate that Tc-99m-Sestamibi (MIBI, DuPont Pharma) is a useful tracer for detecting parathyroid adenomas. We present a patient with focal Tc-99m MIBI uptake in parathyroid carcinoma which has only been described once before (1). Tc-99m-MIBI scintigraphy may be considered for diagnosing pathological parathyroid tissue. But presently the histopathological examination only allows the differentiation between adenoma and carcinoma. PMID- 9394361 TI - Evaluation of pentavalent Tc-99m DMSA scintigraphy in small cell and nonsmall cell lung cancers. AB - AIM: The purpose of this study was to evaluate the clinical usefulness of Tc-99m (V) DMSA in patients suspected of lung cancer and determine whether this agent may have value in differentiation between small cell (SCLC) and non-small cell (NSCLC) lung carcinoma. METHODS: Thirty-six patients with clinical and radiological suspicion of primary lung carcinoma were injected 450-600 MBq of Tc 99m (V) DMSA intravenously. Whole body and planar anterior, posterior thorax images were obtained 4-5 h after injection of the radioactive complex. RESULTS: Histopathological results confirmed 23 NSCLC, 10 SCLC and 1 metastatic lung carcinoma and 2 lung abscess. Nineteen of the 23 (82%) NSCLC and all of the 10 (100%) SCLC cases showed Tc-99m (V) DMSA uptake. Single metastatic lung cancer also accumulated radiotracer. Lung abscess did not show uptake. Lesion/Nonlesion (L/N) ratio of SCLC (1.59 +/- 0.32) and NSCLC (1.43 +/- 0.19) tumour types did not show statistical difference (p > 0.05). Tc-99m (V) DMSA whole body imaging also showed bone metastases. CONCLUSION: Tc-99m (V) DMSA is a noninvasive and cheap imaging method to detect malignant lung cancers and their bone metastases but, differentiation of SCLC and NSCLC is not possible. PMID- 9394363 TI - Clinical experience with gemcitabine in pancreatic carcinoma. AB - The treatment of advanced pancreatic carcinoma has been viewed with pessimism. Because of the lack of activity of commonly used agents, there is no consensus regarding a standard chemotherapy regimen. Assessment of response is neither uniform nor reproducible. Debilitating tumor-related symptoms, including pain, anorexia, weight loss, and impaired performance status, are common. Many studies have failed to evaluate the palliative benefit of treatments, although many patients consider such benefit to be of the utmost importance. Tools have been developed to uniformly assess tumor-related symptoms, and the concept of clinical benefit response has been developed as an end point to quantify symptomatic improvement utilizing these tools. Clinical benefit response incorporates palliative measures, such as pain control, analgesic consumption, performance status, and weight gain. In early phase I and II trials, gemcitabine (Gemzar) has shown activity in patients with chemotherapynaive advanced pancreatic carcinoma. In addition to producing some responses and symptomatic benefit, gemcitabine has a favorable toxicity profile. Two recent trials using clinical benefit response as the primary end point have demonstrated that gemcitabine significantly improves disease-related symptoms in approximately one-quarter of patients. These trials also showed improved survival with gemcitabine, as compared with fluorouracil. Additional studies are required to fully assess the role of gemcitabine in both adjuvant and advanced disease settings. PMID- 9394364 TI - Clinical status and optimal use of topotecan. AB - Topotecan (Hycamtin) is a promising new topoisomerase I-targeting anticancer agent that first entered clinical trials in 1989 under National Cancer Institute sponsorship in collaboration with SmithKline Beecham. In 1996, it was approved for use by the United States Food and Drug Administration (FDA) for previously treated patients with advanced ovarian cancer. For these patients, topotecan provides another therapeutic option upon disease progression after initial platinum-based chemotherapy. Topotecan also has activity in other tumor types, including small-cell lung cancer, hematologic malignancies and pediatric neuroblastoma and rhabdomyosarcoma. Topotecan combination regimens with paclitaxel (Taxol), etoposide (VePesid), cisplatin (Platinol), and cytarabine and with other treatment modalities, such as radiation therapy, are in development. Studies evaluating topotecan combinations as initial treatment in such diseases as ovarian and small-cell lung carcinoma are also underway. It is hoped that earlier use of topotecan, with its novel mechanism of action, will prolong survival and increase cure rates in patients with these chemoresponsive tumors. Whether or not such hopes are realized, these important studies will help define the role of topotecan in cancer chemotherapy. PMID- 9394365 TI - Clinical status and optimal use of the cardioprotectant, dexrazoxane. PMID- 9394366 TI - Clinical trials referral resource. Clinical trials in lung cancer. PMID- 9394367 TI - Anastrozole: a new selective nonsteroidal aromatase inhibitor. AB - Aromatase (estrogen synthetase) is the enzyme complex responsible for the final step in estrogen synthesis--the conversion of androstenedione and testosterone to estrone and estradiol, respectively. Inhibitors of this enzyme have been shown to be clinically effective in the treatment of advanced breast cancer in postmenopausal women, in whom the major source of estrogen production derives from aromatization of adrenal androgens in peripheral tissues, such as muscle, liver, and fat. The most widely used aromatase inhibitor has been aminoglutethimide; however, it is nonselective and also inhibits adrenocorticosteroid synthesis, necessitating hydrocortisone supplementation. Aminoglutethimide is also associated with frequent and troublesome side effects. Formestane, the first selective aromatase inhibitor to be developed, has an improved safety profile and selectivity, but its use has been limited somewhat by its inconvenient administration via intramuscular injection. In this article, the preclinical and clinical data published to date on the new third-generation aromatase inhibitor anastrozole (Arimidex) are presented in the context of current endocrine therapies. Future applications of aromatase inhibitors, both as monotherapy and in combination with other endocrine therapies, are discussed. The use of aromatase inhibitors in advanced disease, the adjuvant setting, and as possible chemopreventive agents are examined. PMID- 9394368 TI - Progress and prospects in vaccine therapy for gynecologic cancers. AB - Therapeutic and prophylactic vaccines that harness the potential of the immune system against a number of gynecologic cancers are now being developed. The therapeutic vaccines coerce the cellular components of the immune system to attack malignant tissue. The prophylactic vaccines induce the production of antibodies capable of neutralizing viral antigens before they infect host cells. However, malignant tumors are usually a heterogeneous mixture of different malignant cells, and it is likely that variant tumor clones within a tumor may not express the target antigen or may possess defects in their antigen-presenting mechanism. Ultimately, therapeutic vaccines may be better suited for the treatment of preinvasive disease or for use as an adjuvant following primary therapy. The prospects for developing efficacious vaccines to treat or prevent cervical, ovarian, uterine, and other gynecologic cancers are promising, however. This article describes the methodology of and rationale for these vaccines. PMID- 9394369 TI - Technology versus biology--where are we headed? PMID- 9394370 TI - Laser biopsy artifact. PMID- 9394371 TI - Transalveolar screw. PMID- 9394372 TI - Pigmented villonodular synovitis of the temporomandibular joint. PMID- 9394373 TI - Ulcerated tumor mass involving the right base of the tongue. PMID- 9394374 TI - Anatomic guidelines for the placement of external references for maxillary repositioning. AB - OBJECTIVE: External reference points, particularly Kirschner pins (K-wire), placed in the region of the nasion have been shown to improve the accuracy of maxillary vertical repositioning. Although no complications associated with this technique have been reported, there is a potential for injury to the anterior cranial fossa or frontal sinus. The purpose of this study was to measure the shortest distance from the nasion to the anterior cranial fossa and from the nasion to the frontal sinus. These measurements were used to establish anatomic guidelines governing safe placement of external reference point pins. STUDY DESIGN: Twenty-seven cadaver heads were sectioned in the midsagittal plane for gross study. Using a Boley gauge, two specific measures were obtained: (1) distance from deepest depression of nasion to the most anterior and inferior projection of the anterior cranial fossa, and (2) distance from nasion to the most inferior aspect of the frontal sinus. All measurements were made in the midsagittal plane. RESULTS: The average distance from nasion to anterior cranial fossa was 16.9 mm (range 13.0 to 20.0 mm) and the smallest distance, 13.0 mm, was seen in two specimens. The average distance from nasion to the frontal sinus was 6.2 mm (range 2.0 to 10.0 mm) and the smallest distance, 2.0 mm, was seen in three specimens. CONCLUSION: Based on our findings, we recommend the following: (1) place pin to a depth of no more than 8 mm into bone, (2) place pin 5 to 10 mm inferior to soft tissue nasion, and (3) place pin in an anterosuperior to posteroinferior direction (i.e., roughly perpendicular to the nasal dorsum). When these anatomic guidelines are followed, one would expect minimal morbidity associated with the placement of ERP pins. PMID- 9394377 TI - Intraoral morbidity following free buccal mucosal graft harvesting for urethroplasty. AB - Buccal mucosa is the preferred donor tissue for urethroplasty in many cases. This study documents donor site morbidity associated with this technique in 12 patients. Nine patients had a transiently decreased parotid salivary flow for 1 week, and one patient reported transient nerve involvement (long buccal nerve). Intraincisal opening returned to presurgical values within 6 weeks and in some cases exceeded presurgical values. This appears to be a low morbidity technique with high patient acceptance. PMID- 9394376 TI - Predictors of postextraction complications in HIV-positive patients. AB - OBJECTIVE: The purpose of this study was to identify factors associated with an increased risk for post-tooth-extraction complications in a sample of HIV positive patients. STUDY DESIGN: A cohort of HIV-positive patients who required the extraction of one or more teeth was enrolled. The predictor variables were grouped into the following sets: demographics; medical and social history; preoperative clinical findings; preoperative laboratory measures (hematologic, immunologic, and nutritional); and treatment. The outcome variable was defined as the presence or absence of a complication following tooth extraction. Logistic regression techniques were used to identify variables associated with an increased risk for complications following tooth extraction. RESULTS: During the enrollment period, 76 HIV-positive patients were enrolled into the study cohort. Seventeen patients (22.4%) had postoperative complications. Based on the bivariate statistical analyses, variables associated with the presence of postoperative complications were red blood cell count, CD8 count, total number of positive sites tested using cell-mediated immunity skin tests, and extraction technique (p < or = 0.05). Using a stepwise logistic regression technique, the variable identified as being predictive of postoperative complications was the CD8 count (p = 0.02). The post-tooth-extraction complication rate of the HIV positive patients in this study sample was greater than the rate reported in most other studies (22% vs. 3%-5%). The complications, however, were minor and easily treated. The variable consistently identified with an increased risk for complications was the CD8 count: the lower the CD8 count, the higher the risk for complications. The CD8 count, however, had poor predictive value. CONCLUSION: In acute clinical situations--for example, in cases of patients with significant dental pain--the results suggest that delaying treatment to obtain laboratory studies may be of little clinical value. It may be appropriate to proceed with suitable, definitive procedure(s) to alleviate symptoms. PMID- 9394375 TI - Radiologic variables of clinical significance in the extraction of impacted mandibular third molars. AB - OBJECTIVE: To determine which radiologic variables have a clinical significance in the extraction of impacted mandibular third molars. STUDY DESIGN: A prospective study was carried out on 100 consecutive extractions of unilateral impacted mandibular third molars (60 women and 40 men, mean age: 26.27 +/- 10.63 years). Fourteen radiologic variables were ordinally evaluated, establishing their relation to the surgical intervention time. The Kruskal-Wallis test, a multivariant analysis of the principal components, the Pearson correlation coefficient, and logistical regression tests were carried out. RESULTS: Seven variables (occlusal plane, relation to the second molar, depth, follicle, periodontal ligament width, ramus of the mandible, and angulation) demonstrated a statistically significant relation to the surgical intervention time (Kruskal Wallis tests, p < 0.007). Two associated variables, depth and periodontal ligament width, showed the most powerful and simple relation to the surgical intervention time (r2 multiple = 0.307, p < 0.001). CONCLUSION: The model we propose is a tool that may help the general practitioner to establish competence in an extraction of the impacted mandibular third molar by measuring the association of two radiologic variables: depth and periodontal ligament width. PMID- 9394378 TI - Preserving the posterior superior synovial recess during allograft TMJ diskal condylar transplantation in the adult goat. AB - OBJECTIVE: The purpose of the study was to test the hypothesis that sparing the posterior superior synovial recess during the resection of temporomandibular joint condyle and disk would maintain a critical mass of synovium necessary to predictably achieve a successful allograft joint reconstruction. STUDY DESIGN: A group of 15 adult goats underwent unilateral resection of their temporomandibular condyle and meniscus. The fossa and posterior superior synovial recess were left intact. They were immediately reconstructed with cryogenically preserved allograft mandibular condyles and temporomandibular joint disk harvested from 15 adult donor goats. The animals were evaluated clinically and radiographically at 6 and 12 months and histologically at 12 months. RESULTS: Of the 15 animals, 13 met all the criteria to be declared a success and retained the posterior superior synovial recess. CONCLUSION: Immediate joint reconstruction using cryogenically preserved mandibular condyles and temporomandibular joint disk can have a high rate of success if the native posterior superior synovial recess remains intact. PMID- 9394379 TI - Kindler syndrome: a rare cause of desquamative lesions of the gingiva. AB - Kindler syndrome is a rare syndrome with cutaneous and intraoral manifestations. It has been suggested that there is an overlap between this syndrome and another called Weary syndrome. Only 68 cases of Weary and Kindler syndromes have been reported, with fewer solely attributed to Kindler syndrome. The salient cutaneous features are neonatal bullae, poikiloderma, photosensitivity, and acral atrophy. This article presents the clinical intraoral findings of two siblings of consanguineous descent diagnosed as having Kindler syndrome. Both had an erythematous and erosive appearance of the gingiva; one sibling had poor oral hygiene and a rapidly progressive form of periodontal disease; the other, whose oral hygiene was acceptable, had no detectable bone loss. PMID- 9394380 TI - Melanocanthoma: a rare cause of oral hyperpigmentation. AB - The oral features of a black woman with melanocanthoma of the oral mucosa are detailed, and the current literature of melanocanthoma of the oral mucosa is briefly reviewed. PMID- 9394381 TI - Spontaneous exfoliation of teeth following severe elemental mercury poisoning: case report and histological investigation for mechanism. AB - BACKGROUND: Although the spontaneous exfoliation of teeth and breakdown of oral tissues from severe mercury intoxication have been noted for over a century, there are no published reports investigating the mechanisms of these phenomena. Severe mercury poisoning is rare in modern times, but it does occur. We present a case report and a histopathologic investigation into the mechanism of the associated tooth loss. METHODS: An exfoliated tooth and periodontal and gingival tissues were obtained from a 15-month-old patient who had been severely intoxicated with elemental mercury over a period of months and hospitalized for severe neurologic and renal effects. The tissues were examined both by routine hematoxylin and eosin stain and by autometallography specific for mercury. For comparison, control tissue from an age-matched subject was examined with the autometallography technique. RESULTS: Under light microscopy, the gingival tissue showed evidence of moderate to severe acute and chronic inflammation. The tooth pulp tissue showed evidence of moderate vascular dilatation and congestion, and it was infiltrated by many neutrophils. The autometallographic sections showed intense accumulations of mercury in the soft tissues of the mercury-exposed subject, but not in the tissues of the control subject. The deposits were primarily found in fibroblasts, which are essential to maintaining the integrity of the oral tissues. CONCLUSIONS: Histopathologic and autometallographic examination of the affected tissue indicates that the primary mechanism of the spontaneous sloughing of tissue and loss of teeth may be the cytotoxic effects of the accumulation of mercury in fibroblasts. Studies of additional cases would be valuable to confirm this hypothesis. PMID- 9394382 TI - An open clinical trial of a new mouth-PUVA variant in the treatment of oral lichenoid lesions. AB - OBJECTIVE: To investigate the feasibility of topical psoralen PUVA (sensitization in photosensitizing psoralen drug + UVA radiation) treatment of oral lichenoid lesions (OLL). STUDY DESIGN: A total of 16 patients with OLL were treated using a 0.01% trioxsalen ointment and UVA doses in the 0.09 to 1.80 J/cm2 range. The average number of sessions was 8.7 and a mean cumulative irradiation dose was 4.25 J/cm2. RESULTS: A marked-to-complete healing occurred in 3 to 16 (19%) patients immediately after therapy, in 4 of 14 (29%) after 3 months, and in 5 of 14 (38%) after 14 months, respectively. Of the 16 subjects with OLL, five were diagnosed as oral lichen planus (OLP) and 11 were classified as oral lichenoid reaction (OLR). Post-PUVA amelioration rate in patients with genuine OLP (4 of 5, 80%) was superior to that in patients with OLR (1 of 9, 11%). CONCLUSION: Topical trioxsalen photosensitization can be used in mouth-PUVA treatment, and lichen planus is a main indication for this therapy. PMID- 9394383 TI - Orofacial pain with vascular-type features. AB - OBJECTIVE: To examine whether a classifiable primary vascular-type craniofacial pain subgroup exists that predominantly affects intraoral structures. STUDY DESIGN: Fifty-five patients were chosen prospectively according to the following inclusion criteria; periodic craniofacial pain that was unilateral, pulsatile, severe, and that may wake the patient from sleep. Accompanying phenomena could include local autonomic and/or systemic signs. Twenty-six cases could be further classified into one of the categories of vascular craniofacial pain. The remaining 29, all with predominantly intraoral pain, were not readily classifiable. RESULTS: Of the 29 patients 70% were women, with an average onset age of 42.6 years. All reported severe, episodic pain that was usually unilateral and lasted minutes to hours. In all, 55% of patients had autonomic or systemic signs, 48% had pulsatile pain, and 35.4% of patients were awakened by the pain. CONCLUSION: Although clinical similarities were observed within these patients, further studies are needed to confirm vascular orofacial pain as a clear diagnostic category. PMID- 9394384 TI - Evaluation of a new device for sterilizing dental high-speed handpieces. AB - Dental high-speed turbines and handpieces can take up and expel microorganisms during operation and thus need regular sterilization. This study established a method for validating devices used to sterilize high-speed turbines and handpieces. The air and water channels and turbine chambers were contaminated with suspensions of Streptococcus salivarius or endospores of Bacillus stearothermophilus. The effect of flushing and/or autoclaving performed by a new device combining both procedures was evaluated by counting the number of viable bacteria recovered from these devices. Further, the effect on clinically used handpieces was evaluated. In an initial experiment, the device partially reduced S. salivarius, and the endospores survived. In a second experiment, a 5 to 6 log reduction of S. salivarius in air and water channels was obtained. No growth was observed in clinically used high-speed handpieces, and both S. salivarius and endospores were eliminated from the turbine chambers. Thus, the method of validation proved capable of discriminating between different levels of bacterial reduction. PMID- 9394385 TI - Mucocutaneous features of autoimmune blistering diseases. AB - This review will describe adult onset mucocutaneous/autoimmune diseases that involve defects in cell-to-cell, cell-to-matrix, or cell-to-basement membrane adhesion. Included in this group are pemphigus, cicatricial pemphigoid, linear IgA bullous dermatosis, epidermolysis bullosa acquisita, and bullous systemic lupus erythematous. Detection and treatment of blistering disorders that manifest early in the oral cavity may prevent widespread involvement of skin. During the past few years, targets of autoantibodies have been clarified and new targets have been identified, allowing better understanding of the pathophysiology involved in these diseases. New information about more effective regimens with fewer side effects has also been obtained, presenting new treatment options. Clinical manifestations and management of these disorders will be described as well as histopathologic, ultrastructural, and immunopathologic studies that distinguish each disorder and facilitate diagnosis and treatment. PMID- 9394386 TI - Keratoameloblastoma of the maxilla. A case report and review of the literature. AB - The keratoameloblastoma is a rare histologic variant of the ameloblastoma. Review of the English language literature revealed five case reports of keratoameloblastoma. We report the sixth case of this tumor. The tumor developed in the right posterior maxilla of a 26-year-old African-American man and demonstrated aggressive clinical behavior, analogous to conventional ameloblastoma. The initial biopsy specimen showed extensive cyst formation, which histologically resembled odontogenic keratocyst. However, the lining epithelium varied in thickness and there was separation and edema between the basal cells and the rest of the epithelium. The basal cells were strongly adherent to the underlying stroma unlike the basal layer of the odontogenic keratocyst, where cleavage often occurs in this area. Additionally, although the basal cells were palisaded and demonstrated nuclear polarization in areas, they were cuboidal rather than columnar. The excision specimen revealed more of a solid ameloblastic component in addition to the cystic component seen on the initial biopsy. Nevertheless, it is possible that the ameloblastoma had developed in an odontogenic keratocyst. Alternatively, it can be postulated that the keratoameloblastoma consists of both cystic and solid components, the former being analogous to the cysts of the conventional ameloblastoma. PMID- 9394387 TI - Distinguishing features of focal cemento-osseous dysplasia and cemento-ossifying fibromas. II. A clinical and radiologic spectrum of 316 cases. AB - The distinguishing histopathologic features of focal cemento-osseous dysplasia (FCOD) (including lesions occurring in both anterior and posterior jaws) and cemento-ossifying fibroma (COF) (ossifying fibroma and cementifying fibroma) were demonstrated in our earlier work. The aim of the current study was to further refine their clinical and radiographic features. We have assessed 18 clinical and radiographic parameters by univariate comparisons (chi-squared and Student t tests), and a multivariate assessment (logistic regression) in 241 cases of FCOD and 75 of COF. These cases were diagnosed from a combination of clinical, radiographic, and histopathologic information. FCOD was seen predominantly in black women, with a peak incidence in the fourth and fifth decades, whereas COF showed no female predilection except in the fourth decade (p < 0.005). COF occurred in patients an average of 10 years younger than patients with FCOD (p < 0.0001). Most patients with FCOD were asymptomatic (62%); the average lesion size was 1.8 cm. More than half of patients with COF displayed jaw expansion and a considerably larger size lesion (mean 3.8 cm, p < 0.001). The mandible was the most frequent site for both FCOD (86%) and COF (70%). Radiographically, a well defined border was observed in 53% of cases of FCOD and 85% of cases of COF (p < 0.01). Cases of FCOD mostly demonstrated an irregularly mixed radio-opacity (69%), whereas 53% of COFs presented as a radiolucency (p < 0.005). In FCOD, there was a close association with tooth apices (70.6%, p < 0.0001) or with previous extraction sites (21%, p < 0.05); however, the majority of COF cases (86%) showed no relationship with either. Combining the radiographic feature of a periapical location with the pathology of multiple curetted fragments and "ginger root" bony trabeculae, allowed 90% sensitivity and 89% specificity in a logistic regression model to predict the lesion to be an FCOD. These findings provide guidelines not only to distinguish these two entities clinically, but also aid in reaching an accurate diagnosis histopathologically. PMID- 9394388 TI - A review of key research design and statistical analysis issues. AB - This article highlights some basic principles of the design and use of statistical tests, using a minimum of mathematics or statistical jargon. It is not the intent to summarize all the possible details involved with performing these tests, but instead to offer insight into evaluating the statistical methods found in published articles. Historically, the number of scientific articles published in which inappropriate statistical analyses were performed is alarming. Only when the reader understands the problem and demands change will this situation improve. The consequence of inaction is to be mired with an array of poorly designed articles that, at the very least, do not advance the field of study and, at worse, may influence practitioners not well versed in statistics to expose patients to useless, unnecessary, or even harmful procedures. PMID- 9394389 TI - Adenomatoid odontogenic tumor presenting as periapical disease. AB - Adenomatoid odontogenic tumor commonly occurs in association with the crowns of unerupted teeth. An extrafollicular variant, radiographically in relationship to root apices, has been reported. However, clear association with the root apices at surgery has not been demonstrated. We report a case of adenomatoid odontogenic tumor in the anterior mandible in a 21-year-old woman that presented radiographically at the root apices and at surgery as a radicular cyst. We believe this represents the first reported case of adenomatoid odontogenic tumor presenting as periapical disease both clinically and radiographically. The diagnosis of adenomatoid odontogenic tumor should be considered when the clinician is presented with a corticated radiolucency in the anterior jaw, especially in teens and young adults. PMID- 9394390 TI - Three-dimensional dental imaging by spiral CT. A progress report. AB - OBJECTIVE: To demonstrate the feasibility of using spiral computed tomographic data for three-dimensional image acquisition, display, and segmentation of dental structures and lesions and to demonstrate the feasibility of metal artifact suppression. STUDY DESIGN: Isolated extracted teeth, a dry mandible, cadaver mandible, and cadaver head were scanned and reconstructed using spiral computed tomography data. Algorithms for metal artifact reduction including extended attenuation range and interpolation of missing projections were applied. Volumetric rendering was performed to synthesize images comparable to conventional intraoral dental radiographs. Serial examinations were obtained by spiral computed tomographic tomography, registered by surface matching, and interval change determined by three-dimensional subtraction. RESULTS: Metal artifact reduction was successful in markedly reducing the streaks and star patterns that usually accompany metallic restorations and intraoral appliances. Voxel sum images were comparable to dental radiographs. Image segmentation could successfully isolate dental structures, and simulated lesions could be detected through three-dimensional subtraction. CONCLUSION: The results demonstrate the feasibility of spiral volumetric computed tomography for quantitative study of oral hard tissues in the presence of metal restorations. PMID- 9394391 TI - Radiographic evaluation of possible etiology of diffuse sclerosing osteomyelitis of the mandible. AB - To examine the cause and site of origin of diffuse sclerosing osteomyelitis of the mandible, we compared various radiographic findings for the mandibular lesions in 20 patients with diffuse sclerosing osteomyelitis with those in 48 patients with osteomyelitis caused by bacterial infection. In osteomyelitis of infectious origin, a typical radiographic feature was a radiolucent lesion spreading in the cancellous bone, with cortical bone perforation and lamellated periosteal reaction. In diffuse sclerosing osteomyelitis, intermingled sclerotic and osteolytic lesions with solid periosteal reaction or external bone resorption were a common finding, and in some patients the cortical bone was initially affected by the fresh or recurrent lesion. Based on these distinct differences, we suggest that the cause of diffuse sclerosing osteomyelitis is not bacterial infection and that the site of origin is not in the bone but in the periosteum. PMID- 9394392 TI - Submandibular gland duct endoscopy. Diagnostic value for salivary duct disorders in comparison to conventional radiography, sialography, and ultrasonography. AB - The purpose of this study is to evaluate the usefulness of endoscopy as a procedure for the diagnosis of submandibular gland duct disorders. Endoscopy of the submandibular glands was performed on 12 patients with symptoms of obstructive sialoadenitis to identify the cause of obstruction. The endoscopic findings were then compared to those of diagnostic procedures such as conventional radiography, sialography, and ultrasonography. Six normal subjects also underwent endoscopy to better understand the normal findings of the duct system. Endoscopy demonstrated salivary gland calculus in 5 of 12 patients, which was revealed as filling defects on sialograms and as strongly echogenic structures on ultrasonograms in 4 of the patients. Endoscopy revealed secretion plugs, secretion plaques, and/or stenosis, which could not be seen by any other diagnostic procedures in 5 patients, as the cause of recurrent swelling in all 7 patients not demonstrating sialolith. Abnormal findings of the duct wall such as vasodilatation, fibrosis, edema, or erythema were seen in four patients, three of whom exhibited dilatation of the duct system on sialograms. In four patients, a decreasing internal echo level of the gland was seen on ultrasonograms. Our initial results for submandibular gland duct endoscopy thus appear to be promising. PMID- 9394393 TI - In vitro magnetic resonance imaging of rodent teeth. AB - OBJECTIVE: The anatomic structure of rat teeth was studied and observed using magnetic resonance imaging with high spatial resolution. STUDY DESIGN: The right part of the low mandible of two rats of 3 and 12 weeks old were analyzed. Images with different orientations were performed in a 2 Tesla magnetic field using the spin-echo imaging technique. RESULTS: Highly spatially resolved images revealed details of teeth, and anatomic differences between a young and an adult rat were demonstrated. CONCLUSION: Magnetic resonance imaging is well suited to image the buccal area and may be a useful tool for the diagnosis of dental diseases. PMID- 9394395 TI - A new ozone-based method for virus inactivation: preliminary study. AB - The nebulization technique reported here could be used to inactivate viruses with ozone in large volumes of body fluids, such as plasma, partial blood and perhaps whole blood in a short time. Coliphage MS2 was used as a model because it is safe, easy to handle and more resistant to chemical disinfections than viruses such as HIV. The theoretical curves and experimental points, describing ozone inactivation of MS2, form a semi-sigmoid of congruent data. There was a > 7log10 reduction in MS2 viability and the possibilities of minimizing the ozone concentration required to kill viruses are indicated. The analysis was expanded to account for the interaction of ozone with a virus suspension in the shape of a thin film from the experimental findings of Bolton et al. We again find a semi sigmoid of congruent data for their case, i.e. describing ozone inactivation of the influenza A virus (WSN strain) and the vesicular stomatitis virus versus time. For the method of nebulization, the exposure time of droplets with ozone is a few seconds, whereas for the thin film method the exposure time is measured in hours. PMID- 9394394 TI - Diffraction enhanced x-ray imaging. AB - Diffraction enhanced imaging is a new x-ray radiographic imaging modality using monochromatic x-rays from a synchrotron which produces images of thick absorbing objects that are almost completely free of scatter. They show dramatically improved contrast over standard imaging applied to the same phantom. The contrast is based not only on attenuation but also the refraction and diffraction properties of the sample. This imaging method may improve image quality for medical applications, industrial radiography for non-destructive testing and x ray computed tomography. PMID- 9394396 TI - Reconstruction of 6 MV photon spectra from measured transmission including maximum energy estimation. AB - Photon spectra from a nominally 6 MV beam under standard clinical conditions and at higher and lower beam qualities have been derived from narrow-beam transmission measurements using a previously published three-parameter reconstruction model. Estimates of the maximum photon energy present in each spectrum were derived using a reduced number of model parameters. An estimate of the maximum contribution of background, or room, scatter to transmission measurements has been made for this study and is shown to be negligible in terms of the quality index and percentage depth-dose of the derived spectra. Percentage depth-dose data for standard beam conditions derived from the reconstructed spectrum were found to agree with direct measurements to within approximately 1% for depths of up to 25 cm in water. Quality indices expressed in terms of TPR10(20) for all spectra were found to agree with directly measured values to within 1%. The experimental procedure and reconstruction model are therefore shown to produce photon spectra whose derived quality indices and percentage depth-dose values agree with direct measurement to within expected experimental uncertainty. PMID- 9394397 TI - Comparison of graphite-to-water absorbed-dose transfers for 60Co photon beams using ionometry and Fricke dosimetry. AB - To derive the absorbed dose to water from a standard of absorbed dose to graphite, the metrology laboratories which apply such a method usually make use of cavity ionization chambers as transfer instruments. In addition, the BNM-LPRI has tested, as such instruments, two types of Fricke dosimeter in its cobalt-60 beam. The two procedures are compared and their results are found to be in good agreement (the difference is less than 0.1%). Both procedures are then taken into account for the calculation of the reference value of absorbed dose to water. PMID- 9394398 TI - Possibilities for tailoring dose distributions through the manipulation of electron beam characteristics. AB - The influence of the properties of an electron beam on resulting dose distributions, and the potential benefits for dose conformity and optimizing dose distribution characteristics by electron beam manipulation, are theoretically examined. A simulated annealing routine is used to weight electron pencil beams of discrete energies incident at discrete locations and angles on one side of a phantom. The resulting optimal electron phase space provides a dose distribution which most closely approaches a desired distribution on the basis of a physical comparison. For simple desired distributions, intuitive results are obtained such as the benefits of energy modulation for distributing dose with depth, of angular and spatial modulation for overcoming disequilibrium effects and their combination in boosting surface doses. For a complex desired dose distribution, the optimization routine instigates a complex interplay of energy, angular and spatial modulation in attempting to achieve dose conformity. A significant result shows that, for a suitably selected beam energy, angular modulation can compensate for the variation in the depth of the distal edge of a superficial target. The effects of varying just energy for normally incident electrons are compared with those of varying the distribution of incidence angle (for monoenergetic electrons) and the combination of both, indicating the relative merits of the manipulation of available degrees of freedom. PMID- 9394399 TI - Optimization of 3D conformal electron beam therapy in inhomogeneous media by concomitant fluence and energy modulation. AB - The possibilities of using simultaneous fluence and energy modulation techniques in electron beam therapy to shape the dose distribution and almost eliminate the influences of tissue inhomogeneities have been investigated. By using a radiobiologically based optimization algorithm the radiobiological properties of the tissues can be taken into account when trying to find the best possible dose delivery. First water phantoms with differently shaped surfaces were used to study the effect of surface irregularities. We also studied water phantoms with internal inhomogeneities consisting of air or cortical bone. It was possible to improve substantially the dose distribution by fluence modulation in these cases. In addition to the fluence modulation the most suitable single electron energy in each case was also determined. Finally, the simultaneous use of several preselected electron beam energies was also tested, each with an individually optimized fluence profile. One to six electron energies were used, resulting in a slow improvement in complication-free cure with increasing number of beam energies. To apply these techniques to a more clinically relevant situation a post-operative breast cancer patient was studied. For simplicity this patient was treated with only one anterior beam portal to clearly illustrate the effect of inhomogeneities like bone and lung on the dose distribution. It is shown that by using fluence modulation the influence of dose inhomogeneities can be significantly reduced. When two or more electron beam energies with individually optimized fluence profiles are used the dose conformality to the internal target volume is further increased, particularly for targets with complex shapes. PMID- 9394400 TI - Correction factors for parallel-plate chambers used in plastic phantoms in electron dosimetry. AB - In electron beam dosimetry using parallel-plate chambers solid phantoms are sometimes necessary. To obtain the dose to water from the ionization obtained in the solid phantom, fluence correction factors and perturbation factors have to be applied. In this study fluence factors in a perturbation free geometry have been determined experimentally for common phantom materials. Wall perturbation factors for simulated Attix, NACP, and Roos chambers have also been determined for the same materials. Comparative Monte Carlo calculations have been performed using the EGS4 Monte Carlo code. Comparison with data in newly published protocols such as IAEA and IPEMB shows an agreement with the results obtained in this paper to within 1%, demonstrating that the data published in these protocols may be used with reasonable accuracy if recommended phantoms are used. The results also show that if unsuitable phantom materials are used, the wall perturbation factors may differ for different chambers and for different phantom materials by more than 3% and perturbation factors have to be considered in order to obtain a high accuracy in the dose determination. PMID- 9394401 TI - Two-dimensional scatter integration method for brachytherapy dose calculations in 3D geometry. AB - In brachytherapy clinical practice, applicator shielding and tissue heterogeneities are usually not explicitly taken into account. None of the existing dose computational methods are able to reconcile accurate dose calculation in complex three-dimensional (3D) geometries with high efficiency and simplicity. We propose a new model that performs two-dimensional integration of the scattered dose component. The model calculates the effective primary dose at the point of interest and estimates the scatter dose as a superposition of the scatter contributions from pyramid-shaped minibeams. The approach generalizes a previous scatter subtraction model designed to calculate the dose for axial points in simple cylindrically symmetric geometry by dividing the scattering volume into spatial regions coaxial with the source-to-measurement point direction. To allow for azimuthal variation of the primary dose, these minibeams were divided into equally spaced azimuthally distributed pyramidal volumes. The model uses precalculated scatter-to-primary ratios (SPRs) for collimated isotropic sources. Effective primary dose, which includes the radiation scattered in the source capsule, is used to achieve independence from the source structure. For realistic models of the 192Ir HDR and PDR sources, the algorithm agrees with Monte Carlo within 2.5% and for the 125I type 6702 seed within 6%. The 2D scatter integration (2DSI) model has the potential to estimate the dose behind high density heterogeneities both accurately and efficiently. The algorithm is much faster than Monte Carlo methods and predicts the dose around sources with different gamma-ray energies and differently shaped capsules with high accuracy. PMID- 9394402 TI - Dosimetry studies with TLDs for stereotactic radiation techniques for intraocular tumours. AB - Between March 1993 and January 1997, stereotactic radiation techniques were used to irradiate 66 intraocular tumour patients with the Gamma Knife (Leksell Gamma Knife, model B unit) at the University of Vienna, Austria. This study investigates the dosimetry for stereotactic irradiation of ocular structures. For the dosimetry program KULA 4.4, Gamma Knife stereotactic irradiation of the eye represents an extreme frontal skull position. In addition, irradiation of the eye may be performed in the usual supine position in exceptional cases only. With the patient in the prone position, the dose planning program has to calculate with a significantly large number of single-beam extrapolations. In our first experiment we measured the isocentre dose for eight different gamma-angle positions, both in prone and supine positions, using TLD measurements in an Alderson head phantom. We found a maximum deviation of +/- 1.6% using these individually calibrated TLDs. In the second experiment we examined the dose cross profiles for the two most frequently used treatment positions (supine position, gamma = 65 degrees, and prone position, gamma = 140 degrees). For this purpose we implanted a specially designed TLD array into the orbit of a human cadaver head. We found excellent agreement of the dose values measured for the isocentre as well as the posterior part of the eye with orbit with deviations of less than -2.7%. However, for the anterior part of the eye, deviations between computer-generated calculations and the TLD measurements were found to range up to -30%. These differences were noticed both for supine and prone positions. For the Gamma Knife stereotactic irradiation of ocular tumours or pathologies, precautions should be taken to avoid significant underdosage in the anterior part of the radiation field. PMID- 9394403 TI - The inverse problem of depth dose curve estimation. AB - The inverse problem of the depth dose curve is formulated and a proposition for its solution is presented. The solution is based on the approximation of the observation equation with a numerical quadrature operator and the regularization of this inverse problem with a smoothness side constraint. The problem formulation is applicable for both the electron and the photon depth dose curve estimation. Moreover, the method is equivalent for, for example, all energies, field sizes and source-to-phantom distances. Simulations show that the estimation error is smaller with the proposed method than with direct linear interpolation. The main result of the paper, however, is the formulation of the problem that allows feasible extensions and modifications for different measurement situations. PMID- 9394404 TI - An automated method for mapping human tissue permittivities by MRI in hyperthermia treatment planning. AB - This paper presents an automatic method to obtain tissue complex permittivity values to be used as input data in the computer modelling for hyperthermia treatment planning. Magnetic resonance (MR) images were acquired and the tissue water content was calculated from the signal intensity of the image pixels. The tissue water content was converted into complex permittivity values by monotonic functions based on mixture theory. To obtain a water content map by MR imaging a gradient-echo pulse sequence was used and an experimental procedure was set up to correct for relaxation and radiofrequency field inhomogeneity effects on signal intensity. Two approaches were followed to assign the permittivity values to fat rich tissues: (i) fat-rich tissue localization by a segmentation procedure followed by assignment of tabulated permittivity values; (ii) water content evaluation by chemical shift imaging followed by permittivity calculation. Tests were performed on phantoms of known water content to establish the reliability of the proposed method. MRI data were acquired and processed pixel-by-pixel according to the outlined procedure. The signal intensity in the phantom images correlated well with water content. Experiments were performed on volunteers' healthy tissue. In particular two anatomical structures were chosen to calculate permittivity maps: the head and the thigh. The water content and electric permittivity values were obtained from the MRI data and compared to others in the literature. A good agreement was found for muscle, cerebrospinal fluid (CSF) and white and grey matter. The advantages of the reported method are discussed in the light of possible application in hyperthermia treatment planning. PMID- 9394405 TI - A study of thyroid radioiodine monitoring by Monte Carlo simulations: implications for equipment design. AB - Monte Carlo simulations have been performed to evaluate the design of collimated detectors used to measure 125I or 131I in the thyroid gland. Two detector sizes were simulated for each radioisotope: (i) for 125I monitoring 2.54 cm diameter and 7.62 cm diameter and 0.2 cm thickness and (ii) for 131I monitoring 2.54 cm diameter, 3.2 cm thickness and 7.62 cm diameter, 6.4 cm thickness. The virtual thyroid gland was 20 g. Activity was placed in both the gland and the remainder of the body in varying amounts to assess the efficacy of collimation. The results show that the detector should be sufficiently large so that its solid angle of acceptance when placed 15 cm anterior to the skin surface will include the whole of a moderately enlarged thyroid gland. Heavy collimation to reduce the contribution of extrathyroidal radioiodine within the subject's body is not normally required. It may be of more value as a positioning device and spacer ensuring an appropriate and constant neck to detector distance than in cutting down counts from extrathyroidal activity. In specifying a sensitive detector system for monitoring intrathyroidal radioiodine, a wide angle of acceptance and sufficient detector crystal thickness take precedence over collimation and shielding. PMID- 9394406 TI - Improving wedged field dose distributions. AB - Dose profiles produced by wedge filters in the non-wedged direction can exhibit a 7% or greater dose reduction at the outer ends of the field compared with open field profiles. However, many planning systems use open field profiles to model wedged dose distributions. In the present work, wedges have been modified to reproduce open field profile shapes. This modification involved removing varying thicknesses of the wedge using a simple milling machine. The wedge thickness was calculated using the assumption that dose is proportional to primary collision kerma. The discrepancies in dose between wedged field and open field profile shapes of up to 7% were reduced to less than 3% with the modifications, even for varying depths and off-axis distances. The necessary measurements are simple to perform, and hence this technique could be applied to improve wedged field dose distributions in other radiotherapy departments. PMID- 9394407 TI - An investigation into the effect of input function shape and image acquisition interval on estimates of washin for dynamic cardiac SPECT. AB - Dynamic cardiac SPECT and PET can be used to measure myocardial perfusion by estimating the kinetic rate constant describing the washing of radioactive labelled tracers from the blood to the extravascular myocardial tissue. Because of differences in photon statistics and data acquisition techniques, protocols which produce optimal estimates of the washin for dynamic cardiac PET may give suboptimal estimates if applied in dynamic cardiac SPECT. Two important factors in the estimation of washin are the shape of the tracer input function and the image acquisition interval. This study uses computer simulations to investigate the effect of varying the tracer infusion length and image acquisition interval on the bias and variance of estimates of washin obtained with dynamic cardiac SPECT and 99mTc-labelled teboroxime. Bias in parameter estimates can be introduced by aliasing, integration of the time-varying radioactivity by the detector, and detector motion. This bias can be reduced by decreasing the acquisition interval and using a longer-duration input function. However, this results in poor photon statistics, which generate large variance, and can also introduce bias in the estimates of the washin. Our studies indicate that better estimates of the washin are obtained by using an acquisition interval that is of sufficient duration to obtain adequate photon statistics even if this is at the expense of temporal resolution. The increase in bias caused by using a 10 or 20 s acquisition interval instead of a 5 s acquisition interval is minimal when compared with the reduction in variance. Variance in estimates is also reduced by using a sharp input function, resulting in higher peak counts during washin. It is also shown that the variance of estimates of the washin increases generally when faster kinetics are observed. This variance can, however, be reduced by using longer acquisition intervals. PMID- 9394408 TI - Noise analysis of MAP-EM algorithms for emission tomography. AB - The ability to theoretically model the propagation of photon noise through PET and SPECT tomographic reconstruction algorithms is crucial in evaluating the reconstructed image quality as a function of parameters of the algorithm. In a previous approach for the important case of the iterative ML-EM (maximum likelihood-expectation-maximization) algorithm, judicious linearizations were used to model theoretically the propagation of a mean image and a covariance matrix from one iteration to the next. Our analysis extends this approach to the case of MAP (maximum a posteriori)-EM algorithms, where the EM approach incorporates prior terms. We analyse in detail two cases: a MAP-EM algorithm incorporating an independent gamma prior, and a one-step-late (OSL) version of a MAP-EM algorithm incorporating a multivariate Gaussian prior, for which familiar smoothing priors are special cases. To validate our theoretical analyses, we use a Monte Carlo methodology to compare, at each iteration, theoretical estimates of mean and covariance with sample estimates, and show that the theory works well in practical situations where the noise and bias in the reconstructed images do not assume extreme values. PMID- 9394409 TI - Pre-processing variance reducing techniques in multispectral positron emission tomography. AB - Stochastic fluctuations and systematic errors severely restrict the potential of multispectral acquisition to improve scatter correction by energy-dependent processing in high-resolution positron emission tomography (PET). To overcome this limitation, three pre-processing approaches which reduce stochastic fluctuations and systematic errors without degrading spatial resolution were investigated: statistical variance was reduced by smoothing acquired data in energy space, systematic errors due to nonuniform detector efficiency were minimized by normalizing the data in the spatial domain and the overall variance was further reduced by selecting an optimal pre-processing sequence. Selection of the best protocol to reduce stochastic fluctuations entailed comparisons between four smoothing algorithms (prior constrained (PC) smoothing, weighted smoothing (WS), ideal low-pass filtering (ILF) and mean median (MM) smoothing) and permutations of three pre-processing procedures (smoothing, normalization and subtraction of random events). Results demonstrated that spectral smoothing by WS, ILF and MM efficiently reduces the statistical variance in both the energy and spatial domains without observable spatial resolution loss. The ILF algorithm was found to be the most convenient in terms of simplicity and efficiency. Regardless of the position of subtraction of randoms in the sequence, reduction of the systematic errors by normalization followed by spectral smoothing to suppress statistical noise produced the best results. However, subtraction of random events first in the sequence reduces computation load by half since the need to pre-process this distribution before subtraction is removed. In summary, normalizing data in the spatial domain and smoothing data in energy space are essential steps required to reduce systematic errors and statistical variance independently without degrading spatial resolution of multispectral PET data. PMID- 9394411 TI - The optical properties of lung as a function of respiration. AB - Lung consists of alveoli enclosed by tissue and both structures contribute to volume-dependent scattering of light. It is the purpose of this paper to determine the volume-dependent optical properties of lung. In vivo interstitial fibre measurements of the effective attenuation coefficient mu eff at 632.8 nm differed during inspiration (mu eff = 2.5 +/- 0.5 cm-1) from that during expiration (mu eff = 3.2 +/- 0.6 cm-1). In vitro measurements on a piglet lung insufflated with oxygen from 50 to 150 ml showed the effective attenuation coefficient at 632.8 nm decreases as a function of oxygen volume in the lung (at 50 ml mu eff = 2.97 +/- 0.11 cm-1, at 100 ml mu eff = 1.50 +/- 0.07 cm-1, and at 150 ml mu eff = 1.36 +/- 0.15 cm-1). This was explained by combining scattering of alveoli (Mie theory) with optical properties of collapsed lung tissue using integrating sphere measurements. Theory and measured in vitro values showed good agreement (deviation < or = 15%). Combination of these data yields the absorption coefficient and scattering parameters of lung tissue as a function of lung volume. We conclude that the light fluence rate in lung tissue should be estimated using optical properties that include scattering by the alveoli. PMID- 9394410 TI - In vitro double-integrating-sphere optical properties of tissues between 630 and 1064 nm. AB - The optical properties (absorption and scattering coefficients and the scattering anisotropy factor) were measured in vitro for cartilage, liver, lung, muscle, myocardium, skin, and tumour (colon adenocarcinoma CC 531) at 630, 632.8, 790, 850 and 1064 nm. Rabbits, rats, piglets, goats, and dogs were used to obtain the tissues. A double-integrating-sphere setup with an intervening sample was used to determine the reflectance, and the diffuse and collimated transmittances of the sample. The inverse adding-doubling algorithm was used to determine the optical properties from the measurements. The overall results were comparable to those available in the literature, although only limited data are available at 790-850 nm. The results were reproducible for a specific sample at a specific wavelength. However, when comparing the results of different samples of the same tissue or different lasers with approximately the same wavelength (e.g. argon dye laser at 630 nm and HeNe laser at 632.8 nm) variations are large. We believe these variations in optical properties should be explained by biological variations of the tissues. In conclusion, we report on an extensive set of in vitro absorption and scattering properties of tissues measured with the same equipment and software, and by the same group. Although the accuracy of the method requires further improvement, it is highly likely that the other existing data in the literature have a similar level of accuracy. PMID- 9394412 TI - An electronic portal imaging device for transit dosimetry. AB - An electronic portal imaging device has been designed and constructed. It consists of an array of 128 CsI scintillation crystals coupled to photodiodes which is scanned across the field in 4 seconds. The linac is operated at a dose rate of 400 cGy min-1 and the dose delivered for image acquisition is approximately 27 cGy. The data acquisition controller is a stand-alone STE computer located within the scan arm. Sample images are presented showing contrast and spatial resolution of the system together with a humanoid phantom image and a clinical image of a breast cancer patient. The phantom images show the detector has a contrast resolution of 0.3% (at 15 mm diameter) and a spatial resolution of 2.5-3.2 mm. Images of uniform Perspex blocks have also been calibrated for thickness, indicating the system can measure radiological thickness to an accuracy of 2-3 mm of water. These results indicate the detector may be used for transit dosimetry applications including compensator design. PMID- 9394413 TI - Dosimetric comparison of an integrated multileaf-collimator versus a conventional collimator. AB - The dosimetric characteristics of both a conventional GE collimator (CC) and a GE multileaf collimator (MLC) are compared for different photon beam energies. The integrated GE MLC consists of 32 pairs of tungsten leaves, replacing the lower pair of jaws of the conventional collimator. Measurements were performed with the conventional collimator before this collimator was replaced by the MLC. All parts of the accelerator except the collimator remained the same. Leakage and transmission measurements show good agreement with the manufacturer's specification, stating a leakage between leaves of less than 1% for all energies and a transmission through leaves of less than 0.5%. The dosimetric characteristics of both collimators are very similar for square and rectangular fields. No significant change in beam quality, beam attenuation and depth of maximum dose could be detected within the measurement accuracy. The MLC output ratio variation is smaller than the one measured with the CC. The penumbra difference in the Y direction is less than 0.5 mm at a depth of 5 cm in phantom; in the X direction the penumbra is 1 mm larger for the MLC due to the rounded leaf fronts. As the two leaf banks replace the lower pair of collimator jaws the distance from the collimator end to the isocentre is similar for the two collimators, therefore the MLC does not reduce the flexibility of the treatment unit. For symmetrical and regular collimator settings the MLC can be treated as the CC. PMID- 9394414 TI - Output ratios in a miniphantom for asymmetric fields shaped by a multileaf collimator. AB - The integrated GE multileaf collimator (MLC) provides the ability to achieve 'double' asymmetric fields: each of the 64 leaves allow an over-axis travel of 10 cm and the Y-jaws allow 20 cm. A formalism has recently been proposed by the authors to calculate the output ratio in a miniphantom for this type of MLC by the product of independent leaf and jaw correction factors. The original proposed formalism was restricted to regular or irregular fields including the collimator rotational axis. Introducing 'reduced coordinates' for the correction factors in the present work this formalism is extended to asymmetric fields where central leaves or jaws overlap the collimator axis. The extended formalism is applied to asymmetric square, rectangular and irregular fields. For all fields checked at a given off-axis position, measured and calculated output ratios agree within 1% for 6, 18 and 25 MV photon beams. To relate output ratios normalized to off-axis points with output ratios on-axis, off-axis ratios are derived from film and miniphantom measurements. Both off-axis ratios agree to within 1% for 6 and 25 MV photon beams; a maximum deviation of 1.3% is observed at 18 MV. Calculated products of output ratios and off-axis ratios derived from films are compared with measurements for asymmetric square, rectangular and irregular fields, and agree mostly within 1% for all energies checked; maximum deviations of 1.3 and 1.6% are observed for 6 and 18 MV photon beams. PMID- 9394415 TI - Use of transgenic and gene-targeted mice to model the genetic basis of hypertensive disorders. AB - As both essential hypertension and hypertension associated with pregnancy (pre eclampsia) have been determined to have strong genetic components, considerable recent research has focused on identifying genes that may predispose to the development of these disorders. Recent advances in molecular genetics and the work of the Human Genome Project have facilitated the identification of genes that may be linked to these hypertensive disorders. Although molecular genetic studies performed in humans and animals can be used to link genes or mutations in genes to hypertension (once identified), studies are needed to assess their biochemical and physiologic importance. In this review, we discuss the ever increasing importance and use of transgenic and gene-targeted mice in modeling the genetic basis of hypertensive disorders. PMID- 9394416 TI - Acute renal failure: growth factors, cell therapy, and gene therapy. AB - The rapid understanding of the cellular and molecular basis of organ function and disease will be translated during the next several decades into new therapeutic approaches to a wide range of clinical disorders, including acute renal failure (ARF). The development of the biotechnology for recombinant genetic engineering has led to the prospect of using purified protein products for therapy. In this regard, the repair of ischemic and toxic ARF is critically dependent on a redundant, interactive cytokine network of growth factors to return kidney function to near normal baseline function. Recombinant growth factors are being tested both experimentally and clinically to accelerate the repair of kidney tissue in this disorder. A newer strategy in biotechnology is the development of cell therapy derivatives. Cell therapy is based on the ability to expand specific cells in tissue culture to perform differentiated tasks and to introduce these cells into the patient either in extracorporeal circuits or as implants as drug delivery vehicles of a single protein or to provide physiological functions. Cell therapy devices are being developed to replace components of renal function that are lost during ARF and chronic renal failure and are not replaced with current hemodialysis or hemofiltration. These new approaches may result in therapeutic modalities that diminish the degree of renal failure and the time needed to recover renal function in acute tubular necrosis. This article examines the future prospects of these developing therapies in the treatment of ARF. PMID- 9394417 TI - Genotyping by PCR-ELISA of a complex polymorphic region that contains one to four copies of six highly homologous human VH3 genes. AB - The Humhv3005 human VH gene is located in an intricate locus that encompasses for each haplotype a combination of one to four copies of six highly homologous VH3 genes. To assess the complexity of this region, we developed a polymerase chain reaction-enzyme-derived immunosorbent assay (PCR-ELISA) method capable of detecting each of the VH3 genes. The method consisted of amplification of selected germline VH3 genes with a biotinylated primer, covalent capture of the amplicons onto streptavidin-coated wells, and quantitative typing of the bound VH3 genes with diagnostic oligonucleotides. Pilot studies of two DNA samples with known presence or absence of hv3005 [according to a characteristic BamH1 restriction fragment-length polymorphism (RFLP)] yielded the expected results. Subsequent analysis of 100 additional DNA samples with the known EcoR1 RFLP of hv3005 showed a complete match between the absence of the 9.4-kb hybridizing band and lack of hv3005-like genes, as determined by PCR-ELISA. Importantly, the PCR ELISA analyses of these 102 genomic DNA samples revealed two new haplotypes in the complex hv3005 region. Combined, these data demonstrate the usefulness and efficiency of this new technique to ascertain the presence or absence of six highly homologous genes in an unusually heterogeneous duplication-insertion deletion region. In the future, a similar strategy may be used to dissect other similarly complex VH genetic loci. PMID- 9394418 TI - Insulin receptor expression and clinical outcome in node-negative breast cancer. AB - The insulin receptor (IR), a ligand-activated tyrosine kinase, is present in breast cancers, but its relationship to patient survival is unknown. The IR was measured in 584 tumor specimens from patients with node-negative breast carcinoma by frozen-section immunohistochemistry and light microscopy. The immunostaining signal was quantitated in relation to both the staining intensity and the proportion of positive malignant epithelial cells. Analyses indicated that patients with tumors with undetectable IR content in malignant epithelial cells (260 cases) had a relatively lower predicted 5-year disease-free survival (DFS) (69% +/- 3%) than did patients with tumors with detectable IR content (324 cases; DFS 76% +/- 3%, p = .032). The significance of IR content in these breast malignant epithelial cells was then analyzed along with patient age, tumor size, progesterone and estrogen receptor status, p53 accumulation, and S-phase. Multivariate analysis of these data revealed that after adjustment for these other variables, IR content was the strongest independent predictive factor for DFS (relative risk = 1.73, p = .005). Interestingly, in a small subset of patients with very high IR content (n = 62), DFS was decreased. These data indicate that IR content in node-negative breast cancers is a significant major predictor of reduced DFS. Moreover, they raise the possibility that the measurement of IR content might provide important information concerning breast cancer biology. PMID- 9394420 TI - The erythropoietin receptor gene is not linked with the polycythemia phenotype in a family with autosomal dominant primary polycythemia. AB - Primary familial and congenital polycythemia (PFCP or familial erythrocytosis) is a rare hematological disorder with either autosomal-dominant inheritance or sporadic occurrence. It is characterized by an increased proliferation of erythroid precursors that results in an elevated red blood cell mass. In some of the PFCP families, the disease phenotype is associated with mutations of the erythropoietin receptor (EPOR). Mutations in other genes are likely to cause PFCP as well, but no evidence so far has been provided to support this contention. In this study, we present a family in which 6 of 15 family members were affected in three generations. We screened exon VIII of the EPOR gene for mutations and found a C-->T substitution (C6148T) in the maternal grandmother of the propositus. The mutated allele of the affected grandmother was not passed to either of her two affected children or to her one healthy child; thus, the disease phenotype was not linked to the C6148T mutation in this family. Further examination of the inheritance of the EPOR gene alleles and sequence analysis ruled out linkage between the disease phenotype and the EPOR gene; therefore, an abnormality in another gene must be the cause of PFCP in this particular family. In three affected family members tested, erythroid progenitors were hypersensitive to EPO. This in vitro behavior of the progenitors confirms the diagnosis of PFCP in these subjects. Moreover, it suggests a dominant lesion of an as-yet unidentified gene, either at the level of the EPOR-signaling pathway or another erythropoiesis regulating pathway that may be responsible for enhanced proliferation of the erythroid progenitors. PMID- 9394419 TI - Media acidification inhibits TGF beta-mediated growth suppression in cultured rabbit proximal tubule cells. AB - Chronic metabolic acidosis induces both hyperplastic and hypertrophic renal growth and is associated with progressive loss of renal function. These studies examine the direct effect of media acidification on the growth of rabbit proximal tubule cells in primary culture. The results demonstrate that media acidification has a direct antiproliferative (hypoplastic) effect on both quiescent and mitogen stimulated [epidermal growth factor (EGF)-stimulated] cells and does not induce hypertrophy. This direct antiproliferative effect of acid is associated with inhibition of EGF-induced phosphorylation of the retinoblastoma protein (pRB), which maintains pRB activity and inhibits cell cycle progression from G1 to S phase. Transforming growth factor-beta (TGF-beta) alone has an antiproliferative effect in these cells. TGF-beta converts EGF-induced hyperplasia to hypertrophy and inhibits EGF-induced pRB phosphorylation. Media acidification inhibits both the antiproliferative effect of TGF-beta and the ability of TGF-beta to convert EGF-induced hyperplasia to hypertrophy. This activity is associated with inhibition of TGF-beta-mediated retention of pRB in the active, hypophosphorylated state. These results demonstrate that metabolic acidosis has a direct growth-suppressive effect on renal epithelial cells but inhibits the growth-suppressive effects of TGF-beta. Inhibition of the antiproliferative effect of cytokines, such as TGF-beta, may be responsible for acidosis-induced hyperplasia in vivo. PMID- 9394421 TI - Structures of RNA-binding proteins. PMID- 9394422 TI - The Hofmeister series: salt and solvent effects on interfacial phenomena. AB - Advances in experimental and computational methodologies have led to a recent renewed interest in the Hofmeister series and its molecular origins. New results are surveyed and assessed. Insights into the underlying mechanisms have been gained, although deeper molecular understanding still seems to be elusive. The principal reason appears to be that the Hofmeister series emerges from a combination of a general effect of cosolutes (salts, etc.) on solvent structure, and of specific interactions between the cosolutes and the solute (protein or other biopolymer). Hence every system needs to be studied individually in detail, a state of affairs which is likely to continue for some time. A deeper understanding of the Hofmeister series can be an extraordinarily valuable guide to designing experiments, including not only those probing the series per se, but also those designed to elucidate the adsorption, aggregation and stabilization phenomena which underlie so many biological events. The aim of this review is to provide an up-to-date framework to guide such understanding, consolidating recent advances in the many fields on which the Hofmeister series impinges. PMID- 9394423 TI - [The impact of conjugal bereavement and the buffering effect of social support on the health of elderly people]. AB - This study examined the impact of the spouse's death on the mental and physical health of the elderly, sixty years and older, and the buffering effect of social support against the impact. A three-year study was conducted of 1,087 people whose spouses were alive at the time of the initial survey. Changes over the three-year period were compared among the following three groups: (1) the spouse died within a year prior to the second survey (N = 21); (2) the spouse died more than a year before the survey (N = 47); and (3) the spouse was still alive (N = 901: the comparison group). Results were as follows: (1) Mental and physical health declined more rapidly in the first group than the comparison group, while no significant change was found for the second group. (2) Social support after the spouse loss significantly helped buffer the negative effect on the mental health, but support prior to the loss had no such effect. Social support had no moderating effect on the physical health. PMID- 9394424 TI - [Multiple goals in conflict resolution: their antecedents and effects upon tactical preference]. AB - The multiple goals theory of conflict management (Ohbuchi & Tedeschi, in press) postulated that participants in a conflict pursue to achieve resource goals (economic and personal resources) and social goals (relationship, identity, justice, and power-hostility). The hypotheses based on this theory were examined by the episode method, in which 207 university students were asked to rate their recent experiences of interpersonal conflicts in terms of participants' attributes, goals, and tactics. More than 80% of the subjects answered that they were motivated to achieve multiple goals in their attempts to resolve the conflicts. Social goals were found to be more strongly activated, and economic resource goals were least strongly activated. Regression analyses revealed that the effects of participants' attributes on tactical preference were mediated by goals. PMID- 9394425 TI - [Does altruism reach beyond the in group?: a social orientation approach]. AB - The experiment of this paper studied the role social orientation would play in double-dilemma situations. In a double-dilemma situation, social dilemmas exist both between and within groups; a cooperation choice at the within-groups level is considered a defection choice at the between-groups level, and vice versa. Using such a situation, whether "others" in other-orientedness are limited to those of the ingroup or include those of a competing group was examined. Each of 132 college students played both an ordinary social dilemma game and a double dilemma game, with equivalent incentive structures. Subjects' social orientation was measured a few days after the experiment. Results indicated that other oriented subjects thought only about ingroup members, and did not care much about the others. Furthermore, social orientation did not affect whether subjects acted similarly or differently for the two dilemma situations. Therefore, social orientation approach to intergroup conflicts apparently had its limitations. PMID- 9394426 TI - [Explanation and estimation of subjective probability by generalized model of Support Theory]. AB - The purpose of this paper is to develop a new model for describing the cognitive process of decision making. Being of a generalization of Tversky and Koehler's Support Theory, the proposed model firstly defines the choice probability among several alternatives in terms of degree of "support." Secondly the model defines the degree of support in terms of objective probability and representativeness. Thirdly, this model specifies the integration process by which one reaches the probability of an event utilizing the already assessed subevents. This model was applied to and tasted by, real experimental data. In the experiment, subjects were asked to evaluate the proportions (a kind of probability) and the degrees of representativeness of two objects. Compared to the estimates derived by a Bayesian approach, the subjective probabilities estimated by the proposed model were shown to be closer to those reported by the subjects. PMID- 9394427 TI - [Relations of "self-oriented perfectionism" to depression and hopelessness]. AB - The purpose of this study was to construct a new multidimensional self-oriented perfectionism scale (MSPS) and to examine the relationship of self-oriented perfectionism to depression and hopelessness in college students. In Study 1, 26 original items of a new MSPS were administered to 132 students and factor analysis revealed 4 solutions: desire for perfectionism (DP), personal standard (PS), concern over mistakes (CM), and doubting of actions (D). Twenty items of the final MSPS had high reliability and validity as an instrument of measuring self-oriented perfectionism (Hewitt & Flett, 1991). In Study 2, 178 students completed a questionnaire consisting of MSPS, stressor scale, depression scale, and hopelessness scale. PS was negatively related to hopelessness, and CM and D were positively related to both depression and hopelessness. Students with high CM scores had higher depression than those with low CM scores, unrelated to the degree of stress. PMID- 9394428 TI - [Effects of self-touching behavior on the performance of lexical retrieval]. AB - In this study, effects of self-touching behavior on the performance of lexical retrieval were investigated. In Experiment 1, 52 women were required to retrieve Japanese idioms, and to recall them approximately 2 minutes after the retrieval. The participants were randomly assigned into two groups; in one group, they were tested with the restriction of their hand movement, whereas in the other group, they were allowed to move their hands freely. Results revealed that when the movement was restricted, their performance in the retrieval task was significantly deteriorated. In Experiment 2, after the presentation of tape recorded verbal stimuli, 26 women were required to recall them either with an interval of 2 minutes or with an interval of 2 weeks. The self-touching behavior was found to occur more often when the recall was performed with the interval of 2 weeks than when it took place immediately after the stimulus presentation. Thus self-touching is considered to serve as a cue to retrieve information stored in the long term memory. PMID- 9394429 TI - [The effect of perceived social support and achievement motive on hopelessness]. AB - The purpose of this study was to investigate the effect of achievement motive on the relationship between perceived social support and hopelessness in elementary school children. Two surveys were administered. The first examined the reliability and validity of an achievement motive scale with 273 4th through 6th graders children. The second examined the joint effect of achievement motive and perceived social support on the tendency to feel hopeless, with 410 children of the same age group. Results confirmed that the two-factor structure was indeed appropriate for an achievement motive scale, and that self-fulfillment achievement motive was a moderating variable of the relationship between perceived social support and hopelessness. PMID- 9394430 TI - [Effects of imagery ability and speech anxiety on imagery vividness of imaginary of speech scene]. AB - Effects of imagery ability and speech anxiety on imagery vividness of imaginary of speech scene were examined. Subjects were divided into four groups in terms of high and low scores of Scale of Mental Imagery-Short Form (SMI-S) and a speech anxiety scale. They imagined themselves in neutral, action and speech scenes. They were asked to rate valence, arousal, and dominance of associated emotion, as well as imagery vividness, of each scene. An SMI-S effect was found on the vividness for neutral and action scenes. For vividness of the speech scene, however, speech anxiety had a stronger effect than imagery ability. The subjects with high speech anxiety significantly decreased imagery vividness, and experienced stronger arousal during imaginary speech. Good-imagery subjects with high speech anxiety reported stronger arousal than poor-imagery subjects. These results suggested that speech anxiety was a major determinant of imagery vividness of imaginary of speech scene. PMID- 9394431 TI - Congenital malformations in experimental diabetic pregnancy: aetiology and antioxidative treatment. Minireview based on a doctoral thesis. AB - Diabetes mellitus in pregnancy causes congenital malformations in the offspring. The aim of this work was to characterize biochemical and morphologic anomalies in the conceptus of an animal model of diabetic pregnancy. In addition, a preventive treatment against diabetes-induced dysmorphogenesis was developed. Congenital cataract was often found in the offspring of diabetic rats. The fetal lenses had increased water accumulation, sorbitol concentration and aldose reductase activity compared to control lenses. The results suggest that the cataracts form via osmotic attraction of water due to sorbitol accumulation in the fetal lens. Another set of malformations, with possible neural crest cell origin, occurred frequently in offspring of diabetic rats. These included low set ears, micrognathia, hypoplasia of the thymus, thyroid and parathyroid glands, as well as anomalies of the heart and great vessels. Furthermore, diabetes caused intrauterine death and resorptions more frequently in the late part of gestation. When the pregnant diabetic rats were treated with the antioxidants butylated hydroxytoluene, vitamin E or vitamin C, the occurrence of gross malformations was reduced from approximately 25% to less than 8%, and late resorptions from 17% to 7%. This suggests that an abnormal handling of reactive oxygen species (ROS) is involved in diabetes-induced dysmorphogenesis in vivo. Indeed, an increased concentration of lipid peroxides, indicating damage caused by ROS, was found in fetuses of diabetes rats. In addition, embryos of diabetic rats had low concentrations of the antioxidant vitamin E compared to control embryos. These biochemical alterations were normalized by vitamin E treatment of the pregnant diabetic rats. The antioxidants are likely to have prevented ROS injury in the embryos of the diabetic rats, in particular in the neural crest cells, thereby normalizing embryonic development. These results provide a rationale for developing new anti-teratogenic treatments for pregnant women with diabetes mellitus. PMID- 9394432 TI - A two-site delfia immunoassay for measurements of the N-terminal peptide of pro atrial natriuretic peptide (nANP). AB - A rapid, sensitive and reliable two-site immunoassay for measurements of the N terminal peptide of pro-atrial natriuretic peptide (nANP) is presented. The method uses one monoclonal antibody, directed against the N-terminal part of nANP, as catcher antibody and another monoclonal antibody, directed against the C terminal part of nANP, as detector antibody. The catcher antibody is biotinylated and is bound to streptavidin pre-coated microtiter strips. The detector antibody is labelled with Europium, which is measured in a Wallac DELFIA time-resolved fluorometer. Blood collected in plain Vacutainer tubes gave same measured amounts of nANP as blood collected in heparinised tubes. Blood collected in tubes containing EDTA gave same measured amounts of nANP as the plain tubes, provided that a 2-step assay protocol was used. Based upon 100 healthy blood donors, a reference interval was calculated to < 450 pmol/L. Within the reference group there was a significant increase of serum nANP with age. Based on 42 patients with different degree of impaired renal function, a significant correlation of nANP and serum creatinine was found. Assay performance, given as total assay variation was 12%, 10% and 9% respectively at serum levels of 140, 970 and 3500 pmol/L. It is concluded that this method is fast, sensitive and reliable for clinical measurements of nANP. PMID- 9394433 TI - Trigger delay in infant ventilators. AB - In an experimental study we determined the response trigger delay time of three infant ventilators with a capacity to detect and support spontaneous breathing. We measured this in anaesthetized cats as the time between the start of phrenic nerve activity and the increase in airway pressure caused by the subsequent inflation. Two modes of ventilatory support were used, namely Assist/Control (A/C) and synchronised intermittent mandatory ventilation (SIMV). We found that ventilators equipped with flow sensors close to the free end of the endotracheal tube had a shorter trigger delay than a ventilator which detected breathing with an abdominal sensor. Further, the trigger delay was shorter in SIMV mode than in A/C mode of operation. A higher set sensitivity reduced the response time. We conclude that triggered ventilation may be used in infants, at least when the spontaneous breathing rate is below 60 breaths per minute. This mode of ventilation could be useful when infants are to be weaned off the ventilator. PMID- 9394434 TI - Clinical characteristics of insulin-dependent diabetes mellitus in children at diagnosis. AB - The clinical characteristics of 60 consecutive children < 16 years in a Swedish county with newly diagnosed diabetes mellitus, are described. Twenty-four of them were 5.0-9.9 years old. The fathers of 12% had diabetes. There was no seasonal variation in the onset of diabetes. Presenting symptoms were polyuria and polydipsia in more than 90% of the cases. School children had a longer duration of symptoms than pre-school children. Most of the children were in a good state of health, and none were unconscious on admission. HbA1C was a good indicator of diabetes duration (R2 = 0.32). Patients with Coxsackie B IgM antibodies had lower blood glucose than those without such detectable antibodies. PMID- 9394435 TI - HLA-A and -B antigens and larynx carcinoma in Greeks. AB - The frequency of HLA antigens in 30 Greek larynx carcinoma patients was more prominent for the A21, A28 and B17 antigens compared to 400 healthy unrelated controls from the same population. It is suggested that immunogenetic factors may contribute to the pathogenesis of this neoplasia. PMID- 9394436 TI - Determination of dissociation constants of loop diuretics in acetonitrile-water mixtures. AB - The dissociation pK values of the representative loop diuretics furosemide, bumetanide and ethacrynic acid in 10, 30, 40, 50 and 70% (w/w) acetonitrile-water mixtures at 298.15 K were determined, according to the rules and procedures endorsed by IUPAC. The variation in pK values over the whole composition range studied can be explained by tacking into account the preferential solvation of ionizable substances in acetonitrile-water mixtures. With a view to determining the pK values of the loop diuretics studied in any of the binary solvent acetonitrile-water mixtures, correlations of pK values and different bulk properties of the solvent were examined, and the linear solvation energy relationships method, LSER, has been applied. The pK values were then correlated with the pi*, alpha and beta solvatochromic parameters of acetonitrile-water mixtures. The resulting equations allowed us to calculate pK values for the loop diuretics in any acetonitrile-water mixture up to 70% (w/w) acetonitrile. PMID- 9394437 TI - Analysis of some herbal plants from India used in the control of diabetes mellitus by NAA and AAS techniques. AB - Elemental analysis of some herbal plants used in the control of diabetes has been done by the techniques of Neutron Activation Analysis (NAA) and Atomic Absorption Spectroscopy (AAS). The elements Mn, Na, K, Cl, Al, Cu, Co, Pb, Ni, Cr, Cd, Fe, Ca, Zn and Hg are found to be present in different plants in various proportions. PMID- 9394438 TI - Multivariate calibration for quantitative analysis of EDXRD spectra from a bone phantom. AB - Phantoms have been constructed to simulate trabecular bone mineral loss and cortical bone thinning which consist of a mixture of hydroxylapatite powder and animal fat in various quantities, surrounded by a varying thickness of dural sleeve. Energy dispersive X-ray diffraction (EDXRD) spectra have been recorded, and multivariate calibration has been performed on the spectra from the bone phantoms. The multivariate technique of partial least squares (PLS) was used to predict the hydroxylapatite content of the phantoms and the dural thickness for measurement times of 250, 50 and 5 s. The calibration phantoms consisted of 10 hydroxylapatite densities ranging from 0.5852 g cm-3 to 0.3703 g cm-3 representing a loss of hydroxylapatite of approx. 40% in 4% intervals. Each phantom had four dural sleeves of thickness 0.5, 1.0, 1.5 and 2.0 mm. Nine test phantoms were constructed with a range of densities that were inside the calibration range. For a measurement time of 250 s the average accuracy of prediction for hydroxylapatite content was approx. +/- 3% while for a measurement time of 5 s this fell to approx. +/- 8%. The dural thickness was predicted to within approx. +/- 0.25 mm for a measurement time of 250 s. The results show that multivariate calibration is a useful technique for obtaining quantitative data of a desired variable from EDXRD measurements which may otherwise be masked or corrupted by other variables. PMID- 9394439 TI - Reinvestigation of a physiological eluate of the 52Fe/52mMn generator. AB - We have achieved a significant step forward in the potential application of 52mMn2+ (T1/2 = 0.35 h, beta + = 97%) as a myocardial imaging agent with positron emission tomography (PET) by the introduction of a 5% (physiological) glucose solution as an eluent for the 52Fe/52mMn generator. Our experiments have demonstrated the favourable properties of a glucose solution with minimal breakthrough (< 0.3%) of 52Fe and yields of up to 90% 52mMn2+. Although it has been shown that lower 52Fe breakthrough is attainable using other eluents, due to the short half life of 52Fe (8.27 h) breakthrough up to 1% would not appear to significantly alter the efficacy of the 52mMn eluted with this 5% glucose solution. The primary advantage of this approach lies in its convenience of application, in that a 5% glucose solution may be administered directly into patients thereby circumventing the major problem of non-injectable eluates previously associated with this generator. PMID- 9394440 TI - A ring model for spatiotemporal properties of simple cells in the visual cortex. AB - A neural model is proposed for the spatiotemporal properties of simple cells in the visual cortex. In the model, several cortical cells are arranged on a ring, with mutual excitatory or inhibitory connections. The cells also receive excitatory inputs either from lagged and nonlagged cells of the lateral geniculate nucleus in one setting or from nonlagged cells in the other. Computer simulation shows that the cortical cells have spatiotemporally inseparable receptive fields in the former setting and separable fields in the latter; spatial profiles at a given time in the spatiotemporal fields are described with a Gabor function whose phase parameter varies regularly from 0 to 2 pi with rotation along the ring; the inseparable cells have directional selectivity as observed physiologically. PMID- 9394441 TI - Episodes of low-dimensional self-organized dynamics from electroencephalographic alpha-signals. AB - Self-organized neuronal dynamics revealed by cortical alpha-rhythms occur as episodes, which are rarely observed without extraction of the alpha-band from the other spectral components. Three episodes of an unusually long duration of 10 s, two with no signal processing after data recording at the clinic, are described and show evidence of low-dimensional alpha-dynamics. The evidence is gained from an analysis of scaled structures appearing in families of slope curves of the correlation integrals and is checked against time reparametrization. The data for the two unprocessed 10-s episodes are used for a test of the methodology, as well as a re-examination of the adequacy of the model of an autonomous dynamic system in steady state and of the concept of an attractor in brain dynamics investigations. Striking evidence for the model's inadequacy is provided by the episode of subject S1. In this example five consecutive overlapping 6-s sections do show evidence for low-dimensional dynamics, whereas the 10-s section containing those sections does not. The episode of subject 1 provides an example of alpha-activity which may involve self-organized dynamics extending down to low frequencies. The system (the neuronal network) showing episodes of attractor ruled dynamics, under conditions of blurred and smoothly fading out evidence that it stays on an attractor, is designated as being ruled by a 'shadow-attractor'. This concept is compared with that of a 'quasi-attractor' introduced by H. Haken in studies of physiological systems. One possible mechanism for the observed episodes is proposed, based on a time-dependent number of enslaved sub-systems. PMID- 9394442 TI - Non-linear analysis of the electroencephalogram in Creutzfeldt-Jakob disease. AB - Creutzfeldt-Jakob disease is a rare, neurological, dementing disorder characterised by periodic sharp waves in the electroencephalogram (EEG). Non linear analysis of these EEG changes may provide insight into the abnormal dynamics of cortical neural networks in this disorder. Babloyantz et al. have suggested that the periodic sharp waves reflect low-dimensional chaotic dynamics in the brain. In the present study this hypothesis was re-examined using newly developed techniques for non-linear time series analysis. We analysed the EEG of a patient with autopsy-proven Creutzfeldt-Jakob disease using the method of non linear forecasting as introduced by Sugihara and May, and we tested for non linearity with amplitude-adjusted, phase-randomised surrogate data. Two epochs with generalised periodic sharp waves showed clear evidence for non-linearity. These epochs could be predicted better and further ahead in time than most of the irregular background activity. Testing against cycle-randomised surrogate data and close inspection of the periodograms showed that the non-linearity of the periodic sharp waves may be better explained by quasi-periodicity than by low dimensional chaos. The EEG further displayed at least one example of a sudden, large qualitative change in the dynamics, highly suggestive of a bifurcation. The presence of quasi-periodicity and bifurcations strongly argues for the use of a non-linear model to describe the EEG in Creutzfeldt-Jakob disease. PMID- 9394443 TI - Manuo-ocular coordination in target tracking. I. A model simulating human performance. AB - During eye tracking of a self-moved target, human subjects' performance differs from eye-alone tracking of an external target. Typical latency between target and eye motion onsets is shorter, ocular smooth pursuit (SP) saturation velocity increases and the maximum target motion frequency at which the SP system functions correctly is higher. Based on a previous qualitative model, a quantitative model of the coordination control between the arm motor system and the SP system is presented and evaluated here. The model structure maintains a high level of parallelism with the physiological system. It contains three main parts: the eye motor control (containing a SP branch and a saccadic branch), the arm motor control and the coordination control. The coordination control is achieved via an exchange of information between the arm and the eye sensorimotor systems, mediated by sensory signals (vision, proprioception) and motor command copy. This cross-talk results in improved SP system performance. The model has been computer simulated and the results have been compared with human subjects' behavior observed during previous experiments. The model performance is seen to quantitatively fit data on human subjects. PMID- 9394444 TI - Manuo-ocular coordination in target tracking. II. Comparing the model with human behavior. AB - Several studies have shown that humans track a moving visual target with their eyes better if the movement of this target is directly controlled by the observer's hand. The improvement in performance has been attributed to coordination control between the arm motor system and the smooth pursuit (SP) system. In such a task, the SP system shows characteristics that differ from those observed during eye-alone tracking: latency (between the target-arm and the eye motion onsets) is shorter, maximum SP velocity is higher and the maximum target motion frequency at which the SP can function effectively is also higher. The aim of this article is to qualitatively evaluate the behavior of a dynamical model simulating the oculomotor system and the arm motor system when both are involved in tracking visual targets. The evaluation is essentially based on a comparison of the behavior of the model with the behavior of human subjects tracking visual targets under different conditions. The model has been introduced and quantitatively evaluated in a companion paper. The model is based on an exchange of internal information between the two sensorimotor systems, mediated by sensory signals (vision, arm muscle proprioception) and motor signals (arm motor command copy). The exchange is achieved by a specialized structure of the central nervous system, previously identified as a part of the cerebellum. Computer simulation of the model yielded results that fit the behavior of human subjects observed during previously reported experiments, both qualitatively and quantitatively. The parallelism between physiology and human behavior on the one hand, and structure and simulation of the model on the other hand, is discussed. PMID- 9394445 TI - Evaluation of a self-consistent method for calculating muscle parameters from a set of isokinetic releases. AB - A new method for calculating parameters describing the force-velocity relationship of the contractile element and the force-extension relationship of the series elastic element of skeletal muscle from a set of isokinetic release contractions is evaluated using experimental and numerical techniques. The method calculates from the set of isokinetic releases those force-velocity and force extension relationships that give a self-consistent description of the data set. The self-consistent calculation method is applied to data obtained from the gastrocnemius medialis muscle of the rat, since for such an animal model both relationships can be independently derived from a set of isotonic release contractions. For the two animals studied, the force-velocity and force-extension relationships calculated by the self-consistent method were in good agreement with the ones derived from isotonic releases performed on the same muscle. The statistical properties of the estimates obtained by the calculation method were investigated using a Monte Carlo technique. The method was found to yield results which were biased by less than 2% and which possessed a coefficient of variation smaller than 5%. These findings indicate that the proposed calculation method can be a useful tool for determining the contractile properties of skeletal muscle as reflected in the force-velocity and force-extension relationships. PMID- 9394446 TI - Nonlinear principal components analysis of neuronal spike train data. AB - Many recent approaches to decoding neural spike trains depend critically on the assumption that for low-pass filtered spike trains, the temporal structure is optimally represented by a small number of linear projections onto the data. We therefore tested this assumption of linearity by comparing a linear factor analysis technique (principal components analysis) with a nonlinear neural network based method. It is first shown that the nonlinear technique can reliably identify a neuronally plausible nonlinearity in synthetic spike trains. However, when applied to the outputs from primary visual cortical neurons, this method shows no evidence for significant temporal nonlinearities. The implications of this are discussed. PMID- 9394447 TI - Modeling neural activity using the generalized inverse Gaussian distribution. AB - Spike trains from neurons are often used to make inferences about the underlying processes that generate the spikes. Random walks or diffusions are commonly used to model these processes; in such models, a spike corresponds to the first passage of the diffusion to a boundary, or firing threshold. An important first step in such a study is to fit families of densities to the trains' interspike interval histograms; the estimated parameters, and the families' goodness of fit can then provide information about the process leading to the spikes. In this paper, we propose the generalized inverse Gaussian family because its members arise as first passage time distributions of certain diffusions to a constant boundary. We provide some theoretical support for the use of these diffusions in neural firing models. We compare this family with the lognormal family, using spike trains from retinal ganglion cells of goldfish, and simulations from an integrate-and-fire and a dynamical model for generating spikes. We show that the generalized inverse Gaussian family is closer to the true model in all these cases. PMID- 9394448 TI - Commentary on the application of (Q)SAR to the toxicological evaluation of existing chemicals. AB - For ethical and financial reasons it is impossible to perform thorough toxicological testing for all of the more than 100,000 substances registered in the European Inventory of Existing Substances. It was therefore investigated whether the application of (quantitative) structure-activity relationships (QSAR) with commercially available computer programs could predict the toxicological profile and help identify those substances requiring priority toxicological testing. Whereas predictions with respect to complex endpoints such as carcinogenicity, chronic toxicity and teratogenicity are still disappointing, more reliable predictions should be forthcoming in the immediate future for sensitisation, mutagenicity and genotoxicity endpoints. PMID- 9394449 TI - The toxic effect of the antibiotic metronidazole on aquatic organisms. AB - The acute toxicity of metronidazole was tested on freshwater and marine organisms. The tests showed effect on Chlorella sp. and Selenastrum capricornutum. 72-hr EC10 of 2.03 mg/l and 19.9 mg/l respectively and 72-hr EC50 values of 12.5 mg/l and 40.4 mg/l respectively were among the results obtained. No acute lethal effect was observed on Acartia tonsa or Brachydanio rerio. The study demonstrates the potential ecotoxic effect of metronidazole, suggesting the need for further investigations of the environmental exposure of medicinal substances. PMID- 9394450 TI - Ela 1.0--a framework for life-cycle impact assessment developed by the Fraunhofer Gesellschaft. Part A: The conceptual framework. AB - The Fraunhofer-Gesellschaft has sponsored the development of a conceptual and flexible, computer aided tool to perform the impact assessment within LCA (life cycle assessment) for technical products and processes. The developed general framework "Ela 1.0" (environmental loads analysis) consists of four elements: the selection of appropriate impact categories, the categorization of emissions and wastes leaving the systems as well as of resource and energy consumption, the characterization and an analysis of the results of the impact assessment. The latter compares the product-based emissions with the total of emissions of a region such as Germany, the EU or OECD countries. The framework Ela 1.0 considers the environmental categories: global warming, ozone depletion, resource and energy consumption, wastes, eutrophication (including COD and BOD as measured parameters), acidification, ecotoxicity, ozone formation and human toxicity. The latter categories are handled by listing of precursors for ozone formation, and by listing of emissions scored according to their human hazard potential. The options, possibilities and limitations of the conceptual framework are presented in part A of a series of publications. PMID- 9394451 TI - Assessment of the environmental behaviour of antioxidants in folios. Comparative risk analysis for the use of folios in agriculture. AB - The objective of the reported study has been to assess and evaluate as comprehensively as possible the environmental impact of Octadecyl 3(3,5-di-tert butyl-4-hydroxyphenyl) propionate (CAS no: 2082-79-3, Irganox 1076, Ciba Specialty Chemicals Inc., Additives), which is used as an antioxidant. The potential impact on the compartments soil, groundwater and surface water is to be considered. For comparative purposes, additionally, other chemical compounds being currently under environmental discussion are also taken into account. These comprise pesticides and phthalates which are ubiquitously distributed plasticizers as well as a complexing agent. Since the data basis for each of the compounds under consideration is different, a tiered approach comprising various methodologies of impact assessment has been chosen to achieve the best possible comprehensiveness. The tiers are: 1. Tier: hazard assessment using a scoring system 2. Tier: comparative risk assessment. When interpreting the results of each method, system boundaries as well as underlying assumptions were taken into consideration. Both methodologies showed, that-as compared to the reference substances-there is no relevant environmental and toxicological concern due to low environmental and human hazard from Octadecyl 3(3,5-di-tert-butyl-4 hydroxyphenyl) propionate. PMID- 9394452 TI - Multinucleated giant cells recruited by implantation of octacalcium phosphate (OCP) in rat bone marrow share ultrastructural characteristics with osteoclasts. AB - The ultrastructural characteristics of multinucleated giant cells (MNGCs) on octacalcium phosphate (OCP) and hydroxyapatite (HA) were investigated in comparison with those of osteoclasts, when the synthetic OCP and HA were implanted in rat bone marrow. The morphological difference in the MNGCs were shown between OCP and HA implants in 2 weeks after implantation. The MNGC on the implanted OCP (i-OCP) developed the ruffled border-like structure and the clear zone-like structure. The i-OCP was frayed where it was in contact with the ruffled border-like structure. The MNGC on the implanted HA (i-HA) developed the clear zone-like structure, whereas no ruffled border was seen. The surface of i HA associated with the MNGC was smooth and not frayed. Bone formed directly on the OCP or HA implants. The interface between the i-OCP and bone matrix interdigitated, whereas that between the i-HA and bone matrix was comparatively smooth. The present study suggested that the i-OCP could be resorbed by the MNGCs which share some ultrastructural characteristics with osteoclasts. PMID- 9394453 TI - Substructures of the acinar basement membrane of rat submandibular gland as shown by alcian blue staining and cryo-fixation followed by freeze-substitution. AB - The ultrastructure of the epithelial basement membrane was studied in the acinar cells of adult rat submandibular glands after: (i) immersion fixation in 0.1 M sodium cacodylate buffer (CB, pH 7.2) containing 2.5% glutaraldehyde (GA) and alcian blue (0.5%) in conjunction with microwave irradiation, (ii) perfusion fixation with 2.5% GA in CB, (iii) rapid freezing followed by freeze-substitution (RF-FS) with 1% GA in acetone, and (iv) RF-FS with 2% OsO4 in acetone. The specimens were post-fixed with 1% OsO4 in CB after methods (i) and (ii) but not (iii) and (iv). Fixed specimens were embedded in epoxy resin and the ultrathin sections were cut, stained with both lead and uranium, and observed under a transmission electron microscope. Various substructural components could be seen in the acinar basement membrane. In the specimens processed by method (iv), a clear meshwork structure could be found just beneath the basal plasma membrane. This meshwork could not be seen in the specimens processed by method (iii) but thin filaments of approximately 100 nm in length extending from the epithelial base toward the connective tissue space were evident. By methods (i) and (ii), an electron dense 30-75 nm layer could be seen subjacent to basal cell membranes. By method (ii), particularly, thick threads connecting this layer to collagen fibres in the connective tissue were stained with alcian blue. Lamina lucida was not identified. PMID- 9394454 TI - Ultrastructural study of capillary and myocytic changes in the masseter and heart of KK-Ay mice. AB - We studied the capillaries and myocytes of masseter and cardiac muscles of diabetic KK-Ay mice, using light microscopy, transmission electron microscopy and scanning electron microscopy. The following changes were observed for diabetic masseters: capillary tortuosity, diversity of capillary caliber, endothelial cell swelling accompanied by luminal narrowing, widening of the pericapillary space and pericapillary fibrosis, and subsarcolemmal accumulation of myocytic mitochondria in areas adjacent to capillaries. In addition, attenuated capillary segments were largely covered by pericytes with profuse processes. In contrast, cardiac muscles of KK-Ay mice exhibited subsarcolemmal accumulation of myocytic mitochondria in areas contiguous to capillaries, and degenerative changes of myocytes such as disarrangement of myofilaments, disappearance of Z-bands and clustering of lipid-like vacuoles, although conspicuous changes of capillaries were not noted. Microvascular and myocytic changes described above may suggest the presence of microangiopathy and myopathy in the masseter and heart of KK-Ay mice. PMID- 9394455 TI - Electron microscopy of biological specimens by the plasma-polymerization rapid freeze replica method. AB - The plasma-polymerization replica method is a unique replica technique for transmission electron microscopy. In the present study, we used this method in combination with a rapid-freeze technique to observe T4 bacteriophages and hepatitis B virus core particles. The heads of T4 bacteriophages appeared hexagonal and measured approximately 110 nm in length. Striations in their tails were also visible, indicating that the resolution of the present method is better than 4 nm. The images corresponded well with those obtained by ice-embedding and negative staining methods, with respect to both morphology and size of the phage particle. Hepatitis B virus core particles observed by the present method appeared round, approximately 30 nm in diameter, with hollow centres. Again, the morphology and size of the particles corresponded well with those obtained by ice embedding, negative staining, and ultrathin sectioning. From these results, we conclude that the plasma-polymerization rapid-freeze replica method provides a useful technique for observation of biological specimens in a natural state and at high resolution. PMID- 9394456 TI - Detection of Epstein-Barr virus DNA in virus-infected cells by electron microscopic in situ hybridization. AB - We describe a procedure for in situ hybridization using a biotinylated Epstein Barr virus (EBV) sequence with detection at the light and electron microscopic levels. In situ hybridization using an immunogold-silver staining detection system was used to identify biotinylated DNA probes in cell smears and in Lowicryl K4M-embedded EBV-infected and -noninfected cell lines. At the light microscopic level, the reaction product of hybridized EBV DNA sequence seemed to be located mainly in the nuclei. The labelling was dependent on the cell strains. However, at the electron microscopic level, the reaction product was evident as spots or clusters distributed not only in the nuclei of EBV-infected cells but also in the cytoplasm and extracellular particles. These findings suggest that immature particles in the cytoplasm contain EBV DNA. This procedure can be applied to the observation and identification of virus infection. PMID- 9394457 TI - Electron microscopic observations of the stichosome during the normal development of Trichinella spiralis from muscle larvae to adult worms in BALB/c mice. AB - The exocrine granules of the stichosome of Trichinella spiralis contain excretory and secretory (ES) products that may alter host cell physiology in such a way that T.spiralis can establish parasitism in the host [1,2]. The stichosome is the most intriguing but still mysterious exocrine organ. This paper describes ultrastructural changes of the stichosome during the normal development from muscle larvae to adult worms. Stichocyte granules of the muscle larva stage were excreted by 14 h post-infection (PI). Then the stichosome synthesized a new type of granules, which disappeared from the stichosome by 30 h PI. These transient granules were morphologically different from granules of muscle larva and adult stages. PMID- 9394458 TI - Ultrastructure of a well-preserved lymphocyte from a mummified human. AB - A well-preserved lymphocyte was found during the electron microscopic examination of the cerebral material recovered from a naturally preserved male mummy from northern Chile dating back over 500 years. The cytoplasmic structures were easily recognizable. This study represents one of the best ultrastructural analyses of mummified human peripheral blood elements. PMID- 9394459 TI - Acquisition of the mental state verb know by 2- to 5-year-old children. AB - The production of the cognitive internal state word know by four 2- to 5-year-old children and their parents was examined. The levels of meaning of cognitive words can be categorized hierarchically along the dimensions of conceptual difficulty and abstractness (see Booth & Hall, 1995). The present study found that children and their parents expressed low levels of meaning less frequently, whereas they expressed high levels of meaning more frequently as a function of age. The children's use of know was also correlated positively with (1) their number of different words produced suggesting that cognitive words are related to more general semantic processes, and (2) with parental use of those same cognitive words suggesting that parental linguistic input may be an important mechanism in cognitive word acquisition. Finally, young children tended to use know more to refer to themselves than to refer to others, whereas their parents tended to use know equally to refer to self and others. The importance of cognitive words in a theory of language acquisition is discussed. PMID- 9394460 TI - Social structure of the mound-building mouse Mus spicilegus revealed by genetic analysis with microsatellites. AB - The Mound-building mouse Mus spicilegus possesses a unique behaviour amongst mice. It constructs large earthen mounds and associated nesting chambers which serve to store food for immature individuals during the winter nesting period. We have used genetic analysis of four autosomal and four X-linked microsatellite loci to determine relationships between individuals inhabiting 40 mounds in Bulgaria. We show that, in almost all cases, individuals in a mound are the product of multiple parentage. We estimate the minimum number of males and female parents contributing offspring to each mound and demonstrate that at least two male and two female parents contribute offspring to a minimum of seven mounds. Analyses of relatedness coefficients and allele sharing values demonstrate that parents of different sibships within mounds are more related than if they had been chosen at random from the population and suggest that it is the female parents that contribute this excess relatedness. These results suggest that the mechanism by which individuals congregate to build mounds is kin-based and that the evolution of mound building and communal nesting in M. spicilegus is due in part to kin selection. This study represents a novel approach to the study of mammalian behavioural ecology. We have used a genetic dataset to construct an outline of social structure in the absence of behavioural data. These inferences can now be used to direct further work on this species. PMID- 9394461 TI - A genomic polymorphism located downstream of the gcvP gene of Escherichia coli that correlates with ecological niche. AB - Current evolutionary theory proposes that niche-adapted microbial populations might evolve through selection for favoured genotypes followed by clonal expansion (Maynard-Smith, 1991). Possible correlations between genomic variation and ecological niche in Escherichia coli isolates derived from human and animal sources were investigated by randomly amplified polymorphic DNA (RAPD) analysis. A 1.6-kb polymorphic marker was identified which was present in 60% of isolates from human clinical specimens but was present in less than 5% of isolates derived from ovine and bovine faeces. The marker maps to a region of the chromosome located immediately downstream from the gene encoding the glycine decarboxylase P protein (gcvP). DNA sequences from marker-positive and marker-negative isolates exhibit an abrupt loss of homology immediately downstream from the transcription termination point of the gene which extends for at least 130-base pairs beyond the gcvP transcription terminator. Sequences spanning this region in marker negative isolates exhibit similarity to the cognate sequence from E. coli K-12, while the corresponding region in marker-positive isolates bears no similarity to any other published sequence. The utility of the marker for investigating the occurrence of human-derived E. coli in the environment was studied in a rural stream. Although the stream carried a high background of animal-derived E. coli, the marker could only be detected in isolates obtained downstream of the human faecal input. The polymorphism therefore shows promise for identification of human-derived E. coli within environments containing isolates from multiple and diverse sources. The methods described here could be used to generate further markers suitable for investigating microbial population ecology. PMID- 9394462 TI - Matriarchal genetic population structure of North American beluga whales Delphinapterus leucas (Cetacea: Monodontidae). AB - The North American beluga whale Delphinapterus leucas population has been divided into a number of putative geographical stocks based upon migration routes and areas of summer concentration. Nucleotide sequences of the mitochondrial DNA (mtDNA) control region were used to assess whether these geographical stocks are genetically distinct. Beluga whale samples from 25 sites were collected primarily from aboriginal subsistence hunts across North America from 1984 to 1994. Thirty nine mtDNA haplotypes were identified in 628 beluga samples. No differences were found in the distribution of haplotypes between male and female beluga whales at any sampling site. These haplotypes segregated into two distinct assemblages in both a haplotype network and a neighbour-joining tree. The haplotype assemblages and a geographically disjunct distribution that suggests postglacial recolonization of the North American Arctic from two different refugia. An analysis of molecular variance based on haplotype relationships and frequency indicated genetic heterogeneity among beluga whale summering groups (P < or = 0.001). Sequence divergence estimates between sampling sites also indicated geographical differentiation, particularly between samples taken at east Hudson Bay or St Lawrence River and the western or central Arctic. The results of this study show a high degree of philopatry to specific summering areas by this highly mobile animal. PMID- 9394463 TI - Intron variation in marbled murrelets detected using analyses of single-stranded conformational polymorphisms. AB - Combination of the targeted amplification of nuclear introns and the analysis of single-stranded conformational polymorphisms has the potential to provide an inexpensive, rapid, versatile and sensitive genetic assay for evolutionary studies and conservation. We are developing primers and protocols to analyse nuclear introns in vertebrates, and are testing them in a population genetic study of marbled murrelets Brachyramphus marmoratus. Here we present protocols and results for introns for aldolase B, alpha-enolase, glyceraldehyde-3-phosphate dehydrogenase and lamin A. Results suggest that this approach presents a potentially powerful method for detecting genetic variation within and among local populations and species of animals: (i) a variety of genes can be surveyed, including genes of special interest such as those involved in disease resistance; (ii) assays are rapid and relatively inexpensive; (iii) large numbers of genes can be assayed, enabling accurate estimation of variation in the total genome; (iv) almost any mutation can be detected in the genes amplified; (v) the exact nature of variation can be investigated by sequence analysis if desired; (vi) statistical methods previously developed for proteins and/or sequence data can be used; (vii) protocols can be easily transferred to other species and other laboratories; and (viii) assays can be performed on old or degraded samples, blood or museum skins, so that animals need not be killed. Results of analyses for murrelets support earlier evidence that North American and Asiatic subspecies represent reproductively isolated species, and that genetic differences exist among murrelets from different sites within North America. PMID- 9394464 TI - Genetic structure of an aphid studied using microsatellites: cyclic parthenogenesis, differentiated lineages and host specialization. AB - In a previous study, samples of the grain aphid Sitobion avenae (F.) were collected from wheat and adjacent cocksfoot hosts in a population thought to be primarily parthenogenetic, and DNA from individual aphids was analysed with a multilocus technique. Here we have applied single-locus microsatellites and a mitochondrial DNA marker to a subset of the same DNA extracts, and have made several additional inferences about important genetic and population processes in S. avenae. Microsatellite analysis indicated very high levels of genic and genotypic variation. S. avenae fell into three genotypic groups inferred to be almost noninterbreeding, while analysis of linkage and Hardy-Weinberg equilibria suggested high levels of sexual recombination within each genotypic group. Host specialization was evident: one lineage was found only on wheat, and one (bearing many alleles inferred to be introgressed from the blackberry-grass aphid S. fragariae (Walker)) was found only on cocksfoot. The third group of interrelated genotypes was found commonly on both hosts. Although most genotypes were found only once, some were much more numerous in the sample than expected from the frequency of the alleles they contained. This, and rapid temporal changes in genotypic composition of samples, indicates strong selective differences between genotypes and lineages. In the major genotypic group, the commonest genotypes were significantly more homozygous than were rare ones: thus these data may help to explain the frequent observation of homozygous excess in aphid allozymes. The genotype group showing S. avenae-like as well as S. fragariae-like alleles also carried S. fragariae-like mitochondrial DNA in at least 25/31 cases, indicating gender-asymmetrical hybridization. PMID- 9394465 TI - Techniques for application of faecal DNA methods to field studies of Ursids. AB - We describe methods for the preservation, extraction and amplification of DNA from faeces that facilitate field applications of faecal DNA technology. Mitochondrial, protein encoding and microsatellite nuclear DNA extracted and amplified from faeces of Malayan sun bears and North American black bears is shown to be identical to that extracted and amplified from the same individual's tissue or blood. A simple drying agent, silica beads, is shown to be a particularly effective preservative, allowing easy and safe transport of samples from the field. Methods are also developed to eliminate the risk of faecal DNA contamination from hair present in faeces. PMID- 9394466 TI - Developing microsatellites when they are rare: trinucleotide repeat loci in the northern mockingbird Mimus polyglottos. AB - The great value of microsatellite loci to population studies spurs their development. However, finding loci is difficult when microsatellites are uncommon in the genome. Because trinucleotide-repeat loci are rare in northern mockingbirds Mimus polyglottos, we sought a new method for developing a suite of loci. Here we show that a bacterio-phage cloning vector, Lambda Zap Express (Stratagene, La Jolla, CA, USA) has several features which make it suitable for this purpose. Using this vector, we made a library of 150,000 size-selected clones and screened with an AAT10 probe; 97 positives were identified. From these, 12 pairs of PCR primers were developed, nine of which amplify polymorphic loci. Certain combinations of these primer pairs enable simultaneous amplification of up to three loci. PMID- 9394467 TI - Characterization of microsatellite loci for a co-operatively breeding honeyeater. PMID- 9394468 TI - Learning in the development of infant locomotion. AB - Infants master crawling and walking in an environment filled with varied and unfamiliar surfaces. At the same time, infants' bodies and skills continually change. The changing demands of everyday locomotion require infants to adapt locomotion to the properties of the terrain and to their own physical abilities. This Monograph examines how infants acquire adaptive locomotion in a novel task- going up and down slopes. Infants were tested longitudinally from their first week of crawling until several weeks after they began walking. Everyday locomotor experience played a central role in adaptive responding. Over weeks of crawling, infants' judgments became increasingly accurate, and exploration became increasingly efficient. There was no transfer over the transition from crawling to walking. Instead, infants learned, all over again, how to cope with slopes from an upright position. Findings indicate that learning generalized from everyday experience traveling over flat surfaces at home but that learning was specific to infants' typical method of locomotion and vantage point. Moreover, learning was not the result of simple associations between a particular locomotor response and a particular slope. Rather, infants learned to gauge their abilities on-line as they encountered each hill at the start of the trial. Change in locomotor responses and exploratory movements revealed a process of differentiation and selection spurred by changes in infants' everyday experience, body dimensions, and locomotor proficiency on flat ground. PMID- 9394470 TI - Towards a chemical etiology of nucleic acid structure. AB - In the sequel of some general remarks on a chemical etiology of nucleic-acid structure, the paper presents a reproduction of the sequence of slides which were shown in the author's lecture 'Pyranosyl-RNA' at the 8. ISSOL Conference in Orleans. Each slide figure is accompanied by a short explanatory comment. PMID- 9394469 TI - 30 years later--a new approach to Sol Spiegelman's and Leslie Orgel's in vitro evolutionary studies. Dedicated to Leslie Orgel on the occasion of his 70th birthday. AB - The conditions necessary for evolution are amplification, mutagenesis and selection. Here we describe the evolutionary response of an in vitro replicating system to the selection pressure for fast growth and show what happens to the amplified molecules within this replication system. Our emphasis is on methodology, on the monitoring and the automation of experiments in molecular evolution. In order to perform in vitro studies on the evolution of RNA molecules, a modified self-sustained sequence replication (3SR) method was used. In the first step of the 3SR reaction, the RNA template is reversely transcribed by HIV-1 reverse transcriptase, followed by a second strand synthesis and the transcription of the resulting dsDNA by T7 RNA polymerase. The selection pressure (fast growth) was achieved by applying the principle of serial transfer pioneered in the laboratories of Sol Spiegelman and Leslie Orgel. At the end of the exponential growth phase of the 3SR reaction, an aliquot of the reaction mixture is transferred into a new sample containing only buffer, nucleotides and enzymes while RNA template molecules are provided by the transfer. The conditions in the exponential growth phase allow the RNA molecules to be amplified in a constant environment; all enzymes (HIV-1 reverse transcriptase and T7 RNA polymerase) and nucleotides are present in large excess. Therefore, transferring reproducibly within the exponential growth phase is equivalent to selecting for fast growth; those molecules which can replicate faster will displace others after several transfers. The experiments were performed using a serial transfer apparatus (STA) which allows the nucleic acid concentration to be monitored on-line by measuring the laser-induced fluorescence caused by intercalation of thiazole orange monomers into the RNA/DNA amplification products. The serial transfer experiments were carried out with an RNA template (220b RNA) that represents a 220-base segment of the HIV-1 genome and comprises the in vivo primer binding site (PBS) for the HIV-1 reverse transcriptase. It could be shown that after only two serial transfers two RNA species (EP1 and EP2) emerged that were much shorter. EP1 (48b) and EP2 (54b) were formed by deletion mutations within the original 220b RNA template in the very beginning of the serial transfer experiment; due to their higher replication rate (calculated from the growth curves derived on-line) these two deletion mutants displaced the original 220b RNA template in the course of the following thirty transfers. We assume that these two RNA species evolved independently of each other. Their formation was probably induced by a strand transfer reaction of HIV-1 reverse transcriptase. Sequence analyses of these two evolution products seem to confirm such a presented pathway. 30 years after Spiegelman's experiment, the study described here is another answer to the question he posed: 'How do molecules evolve if the only demand is the biblical injunction: multiply?'. The answer, derived from a modified 3SR amplification system (mimicking a part of the HIV-1 replication cycle in vitro), is the same as thirty years ago: The RNA molecules adapt to the new conditions by throwing away any ballast not needed for fast replication. Clearly, this is only one aspect of molecular evolution; however, it shows that we should be careful in designating unidentified genetic material as 'junk DNA'. PMID- 9394471 TI - Four-stranded DNA formed by isoguanine quartets: complex stoichiometry, thermal stability and resistance against exonucleases. AB - Single stranded DNA-fragments containing short runs of isoguanine such as d(T4iG4T4) (5) or d(iG4T4) (6) form quartet structures by self-assembly of the isoguanine residues. The stoichiometry of the complexes is deduced from mixed aggregates formed between d(T4iG4T4) and d(iG4T4). The iGd-tetrads are more stable with regard to their thermal denaturation and to their resistance against enzymatic phosphodiester hydrolysis than those formed by dG. PMID- 9394472 TI - The evolutionary design of error-rates, and the fast fixation enigma. AB - Genetic and non-genetic error-rates are analyzed in parallel for a lower and a higher organism (E. coli and man, respectively). From the comparison of mutation with fixation rates, contrasting proposals are made, concerning the arrangement of error-rates in the two organisms. In E. coli, reproduction is very conservative, but genetic variability is high within populations. Most mutations are discarded by selection, yet single mutational variants of a gene have, on average, little impact on fitness. In man, the mutation rate per generation is high, the variability generated in the population is comparatively low, and most mutations are fixed by drift rather than selection. The variants of a gene are in general more deleterious than in E. coli. There is a discrepancy in the published mutation rates: the rate of mutation fixations in human populations is twice or four times higher than the individual rate of mutation production, a feature which is not consistent with current population genetics models. Two, not mutually exclusive, hypotheses may explain this 'fast fixation enigma': (i) Mutation rates have substantially decreased in recent human evolution and (ii) A substantial fraction of the fixed mutations were generated in a process-such as gene conversion-that violates the principle of independence of mutation events. PMID- 9394473 TI - Melatonin: pathway from obscure molecule to international fame. PMID- 9394474 TI - Why I changed my mind about water fluoridation. PMID- 9394475 TI - On the causation of edema: a lymphologic perspective. PMID- 9394476 TI - Emerging pathogens: threat and opportunity. PMID- 9394477 TI - Rene J. Dubos and Fred L. Soper: their contrasting views on vector and disease eradication. PMID- 9394478 TI - Levels of heavy metals in seals of Lake Ladoga and the White Sea. AB - Between 1990 and 1993 samples of hair, liver, kidney and muscle were collected from 28 ringed seals from Lake Ladoga, Phoca hispida ladogensis, 20 ringed seals, Phoca hispida hispida, and three bearded seals, Erignathus barbatus, from the White Sea for heavy-metal residue analyses in tissues. The concentration of Hg, Cd, Pb, Cu, Ni and Zn were determined by atomic absorption spectrometry (AAS). The samples of hair and liver contained the highest mean levels of the elements analysed and the muscle contained the lowest mean heavy-metal concentrations. Age and sex differences in the accumulation of pollutants were found. Tissues of Ladoga ringed seal were to a greater extent contaminated with the heavy metals studied than the tissues of the White Sea pinnipeds. PMID- 9394479 TI - Angular and fibrous particles in lung are markers of job categories. AB - INTRODUCTION: The lung concentration of angular and fibrous particles has been measured when cases are stratified into their job categories; 21 miners (metallic mines such as gold, zinc and copper), 18 iron foundrymen, 22 non-iron foundrymen, four welders, three sand-blast workers, four construction workers, three technicians and professionals, seven workers in other trades excluding welding. Twelve asbestos miners representing a positive exposure to asbestos and 20 people representing a background population were added to the previous groups. MATERIAL AND METHODS: Particles, both angular and fibrous, were extracted from lung parenchyma by a bleach digestion method, mounted on copper microscopic grids by a carbon replica technique and analyzed by transmission electron microscopy (TEM) and energy dispersive spectroscopy (EDS). Quartz concentration was also determined by X-ray diffraction (XRD) on a silver membrane filter after the extraction from the lung parenchyma. RESULTS: (1) The highest concentrations of quartz were found in mines (metallic mines), iron foundrymen and sand-blast workers. Notable amounts quartz were found in welders and professionals. (2) The highest concentrations of short fibres were found in non-iron foundrymen, asbestos miners and construction workers. (3) The highest concentrations of long fibres were found in non-iron foundry men and asbestos miners. (4) The highest concentrations of ferruginous bodies were found in non-iron foundrymen and asbestos miners. (5) The non-iron foundrymen were exposed to ceramic fibres and asbestos fibres. CONCLUSION: The results of the study may not be representative of the broad spectrum of workers in the industrial activities in which they have been involved. However, the detailed composition of the retained particles of our workers is explained both qualitatively and quantitatively by their work histories. Finally, the broad range of particle types identified in the lungs of these workers illustrate the complexity or trying to determine disease origins in these occupational settings. PMID- 9394480 TI - A model of the distribution and retention of tungsten in the human body. AB - Expanding industrial and military uses of tungsten could result in substantially increased levels of this metal in the environment in the next few years. Although occupational experiences and available toxicological studies on laboratory animals suggest that tungsten may have a relatively low order of toxicity, the data are weak and inconclusive. There is a need not only for more systematic studies of the behavior and effects of tungsten in different animal species but also for a reliable, biologically realistic biokinetic model for tungsten in man that can be used to relate concentrations of this metal in environmental media to concentrations in tissues of exposed persons and translate results of experimental studies into terms of environmental exposures. This paper is intended as a first step toward development of such a biokinetic model. Information related to the biokinetics of tungsten in mammalian species is examined, a biologically meaningful compartmental model structure is proposed, provisional transfer rates between compartments are selected, areas are identified where additional biokinetic data on tungsten are most needed and suggestions are made for further research into the biokinetics of tungsten. PMID- 9394481 TI - Modeling methane emissions from cattle in Mexico. AB - Modeling methane emissions from cattle requires data on herd size, herd distribution by weight and use, and distribution by climate. In this article, it is shown how empirical and semi-empirical models were obtained for these data in Mexico. Some shortfalls in the Tier 2 approach of the 1994 IPCC's methodology for emissions from enteric fermentation are discussed and an intermediate procedure is proposed. These methods could also be applied in other countries. PMID- 9394482 TI - Environmental pollution and child health in the Aral Sea region in Kazakhstan. AB - The deterioration of human health with increasing infant mortality rate, declining life expectancy at birth and increasing prevalence of serious infectious diseases in Russia and other former Soviet Republics is thought to be due to a combination of several factors such as inadequate nutrition, poor sanitation, collapse of the health care system and pollution from Soviet agriculture and industries. In the Aral Sea region in Kazakhstan, the environmental problems are of near catastrophic proportions. As a result of the implementation of a massive irrigation scheme to support the cotton fields in the former desert land, the water flow to the Aral Sea was reduced to less than half. Industrial pollutants such as PCB-compounds and heavy metals, but also the use of large quantities of pesticides to control parasites and weeds have accumulated not only in water, but also in soil and have been deposited over large areas by atmospheric transport to enter the food chain leading to humans. In a study of 15 children and of an additional 12 children referred from the region of the Aral Sea to the National Children's Rehabilitation Center in Almaty with symptoms and signs of 'ecological disease', we have found that the concentration of PCB compounds in the blood lipids is elevated in relation to healthy Swedish children. In addition, the blood lipid concentration of the beta-isomer of the hexachlorocyclohexanes was extremely high and of DDT-compounds was elevated up to 20 times. The concentrations of lead in red blood cells was moderately elevated and that of cadmium slightly elevated compared to the findings in Stockholm children. To study the role of these pollutants in the diseases found in children from the Aral Sea region accurate epidemiological studies have to be performed. PMID- 9394483 TI - Response of vehicular lead to the presence of street dust in the atmospheric environment of major roads. AB - Size fractionated particulate samples were collected from the roadside atmosphere of three major roads within the Brisbane Metropolitan area, using a high volume sampler fitted with an Anderson impactor. Street dusts were also sampled at these sites. Deposition samples were collected simultaneously with those of atmospheric particulates from periods with and without rainfall. All types of samples were quantitatively analysed for lead and various anions and cations. The pH and electrical conductivity for street dusts and deposition samples together with total solids content of deposition samples were also determined. Results showed that at sites where the process of street dust resuspension was at a minimum, the bromide-to-lead ratios were comparable to the reported ratio in uncombusted petrol. However, the relatively higher bromide-to-lead ratios observed at sites with active street dust resuspension indicate the existence of a process by which fine lead particulates are removed from the atmosphere by resuspended coarse dust particles. PMID- 9394485 TI - Disposal of chemical weapons in the Baltic Sea. AB - Large quantities of chemical warfare agents were dumped in the Baltic Sea after World War II (WWII). This included 32,000 t of chemical munitions containing approximately 11,000 t of chemical warfare agents which were dumped into the Bornholm Basin and 2000 t of chemical munitions containing approximately 1000 t in the Gotland Basin. Because this material was contained in wooden crates, it was distributed throughout the Baltic. The long-term environmental impact of these agents is unknown. PMID- 9394484 TI - Lead and fluoride content in human bone and hair in the Gdansk region. AB - The post-mortem lead and fluoride content in the rib bone and hair of 59 persons from the Gdansk region was determined. Fluoride was estimated potentiometrically using a fluoride-specific electrode. Lead was determined by atomic absorption spectrometry. The mean lead concentration in bone was 3.0 micrograms/g and 5.2 micrograms/g in hair. The mean fluoride concentrations were 625.7 micrograms/g and 1.4 micrograms/g in bone and hair, respectively. A positive correlation between the lead content in bone and hair was found in this study (r = 0.308, P < 0.05), no such correlation was observed between the fluoride content in bone and hair. A positive correlation between the fluoride and lead contents in bones and the age of the investigated persons was found. The results obtained correspond well with the comparably moderate exposure to lead and fluoride in the Gdansk region. PMID- 9394486 TI - Neurotransmitter transporters: new insights into structure, function and pharmacology. AB - Neurotransmitter transporters on neurons and glial cells catalyze the uptake of neurotransmitter, and may serve to limit the activation of receptors during synaptic signaling. Over the past few years significant progress has been made toward a molecular understanding of neurotransmitter transporters in the CNS. The plasma membrane neurotransmitter carriers are comprised of two structurally- and mechanistically-distinct gene families, the Na+ and Cl(-)-dependent transporters that include the carriers for most of the classical CNS neurotransmitters and several additional carriers for amino acids and other substrates outside the nervous system. A second structurally distinct family of Na(+)-dependent carriers encompasses the excitatory amino acid transporters. For both carrier families the transport of substrate is coupled to the cotransport of sodium ions down a concentration gradient. Electrophysiological studies of neurotransmitter transporters indicate that many of the carriers are electrogenic and may operate in some ways similar to ion channels. In addition, emerging data indicate that these carriers not only function in the uptake of neurotransmitter, but also that as a consequence of their ability to alter the membrane potential they may have a broader role in regulating neuronal excitability and signaling mechanisms. PMID- 9394487 TI - Developmental strategies underlying the elaboration of cortical circuits. AB - The mammalian cerebral cortex is organized in layers and columns, which are reflected in the local intrinsic connections and in the projections to and from the cortex. It is well established that the development of the columnar architecture is under the influence of neuronal activity, but little is known about the mechanisms that control the laminar specificity of cortical circuits. Here we review some recent studies which show that diffusible and membrane associated molecules provide sufficient information to reconstruct layer-specific intrinsic and extrinsic cortical circuits under in vitro conditions. PMID- 9394488 TI - Heterogeneity of median and lateral midbrain radial glia and astrocytes. AB - In the developing mammalian midbrain, radial glial cells are divided into median formations and lateral radial systems with differential properties including rate and timing of cell proliferation, expression of cytoskeletal and calcium-binding proteins, storage of glycogen and relations to afferent fiber systems. To test the hypothesis that radial glial cells of median and lateral midbrain sectors and/or their derivatives are heterogeneous in their relations with local neurons, an in vitro system has been developed and has also been characterized in terms of extracellular matrix (ECM) components. Confluent astrocyte cultures, derived from median (M) or lateral (L) embryonic mouse midbrain sectors, were used as substrates for culturing dissociated cells from median (m) or lateral (l) sectors of embryonic midbrains. In spite of the morphological invariance of glial substrates at confluency, cells that were plated onto these substrates and that were immunoreactive for neuronal markers (MAP2, polysialylated N-CAM or beta III tubulin) showed differences in the aggregation of somata and in the length, caliber and branching of neurites. These differences, which depend mostly on the sector of origin of astrocytes (L: permissive, M: non-permissive for neuronal growth), suggest that the substrates may differ in adhesiveness and/or their carrying of growth-promoting vs. growth-interfering molecules. Indeed, L and M cultures differ in laminin deposition patterns (L: fibrillar, M: punctate pattern). Furthermore, sulfated glycosaminoglycans (s-GAGs) isolated from the pericellular (P), intracellular (I) and extracellular (E) compartments of these sectoral cultures also showed correlations with the ability to support neurite growth. The total amount of s-GAGs in M cultures was twice that in L cultures and was particularly high in the P compartment, with about 3 times as much heparan sulfate (HS) and about 15 times as much chondroitin sulfate (CS) in this fraction of M than in the corresponding compartment of L glia. Our results indicate that cultured astrocytes have heterogeneous properties including different organization of their extracellular matrix that reflect the roles played by their parent radial glia in regions favorable to axonal growth or barrier regions of the developing brain. PMID- 9394489 TI - Macroglia cells of the macaque monkey retina. AB - There are two types of macroglia cells in the macaque monkey retina: Muller cells and astrocytes. Both cell types are in close contact with neuronal structures as well as with the retinal vasculature and are thus well suited for their many physiological tasks. Muller cells ubiquitously traverse the whole thickness of the retina whereas astrocytes are only found in the ganglion cell and nerve fiber layers of vascularized retinal regions. In the adult, astrocytes are very scarce in the central 4 mm around the fovea, a region coinciding with peak Muller cell densities. During development this area is transiently occupied by astrocytes which then disappear during the first postnatal weeks at least in part through apoptosis. Possible reasons for this transiency will be discussed. PMID- 9394490 TI - Regeneration of cat's optic nerve and its functional recovery. AB - The optic nerve of adult mammals can regenerate when a permissive environment is provided with a peripheral nerve (PN) graft. Using this method of PN transplantation, we have studied regeneration of the optic nerve in adult cats. Number of retinal ganglion cells (RGCs) which regenerated their axons through the PN graft corresponds to 3-4% of the total RGC population. The RGCs with regenerated axons distributed widely from central to peripheral retinas. Of the known cell types of cat's RGCs, alpha, beta, gamma and other cells, alpha cells revealed the greatest capacity to regenerate their axons. Dendritic field diameters of most RGCs with regenerated axons were preserved. These regenerated axons were, however, mostly unmyelinated when surveyed by electron microscopy at two months after the transplantation surgery. The regenerated axons revealed normal physiological properties in response to visual stimuli and were classifiable into Y, X or W cells. In accordance with morphological data, Y cells (morphological alpha cells) were more frequently sampled than in normal retinas, whereas the occurrences of X cells (morphological beta cells) and other cells were unchanged or decreased. These results suggest that RGCs retain their physiological function during axonal regeneration, and RGCs with large soma and large dendritic field (Y or alpha cells) have the greatest capacity to regenerate their axons. PMID- 9394491 TI - Natriuretic peptides in the rat olfactory system. AB - The olfactory system plays an important role in the mobilization of animal behaviour, along with the sense of taste. These functions are mediated by a complex olfactory structure composed of peripheral (olfactory epithelium) and central (olfactory bulb) components. Several neuropeptides are synthesized in the olfactory system and are believed to be involved in olfactive processing. Recently, another bioactive substance, atrial natriuretic peptide (ANP), has been demonstrated in the olfactory system. ANP is a potent diuretic, natriuretic and vasorelaxant hormone which, originally, was isolated from mammalian atria, but its gene is expressed in many loci. ANP is only one member of the natriuretic peptide (NP) family, which includes two other peptides, BNP (brain natriuretic peptide) and CNP (C-type natriuretic peptide) derived from different genes. All three peptides show many common features. A high concentration of ANP immunoreactive varicose fibers has been detected in the rat olfactory nerve layer and glomerular layer of the olfactory bulb (OB). An important group of perikarya and thin varicose fibers has been observed in the olfactory tubercle. We have demonstrated the presence of both ANP precursor and ANP gene transcript in the rat olfactory bulb. In addition to synthesizing ANP, the OB contains ANP transducing receptors coupled to guanylyl-cyclase system. The immunoreactive ANP has also been detected in the rat olfactory mucosa (OM), where ANP has been localized in secretory cells of Bowman's gland and in the cells of the epithelial layer. The relatively low concentration of ANP in OM (2.5 ng/mg protein) suggests a local role for ANP, a hypothesis which is strengthened by the presence of ANP high affinity receptors in this tissue. Although the role of ANP in the olfactory system is not yet clear, ANP has been shown to modulate olfactory bulb mitochondrial membrane activity and to be involved in the olfactive function. PMID- 9394492 TI - Death in a dish: controls of apoptosis within the developing retinal tissue. AB - Studies of programmed cell death in the developing retina in vitro are currently reviewed. The results of inhibiting protein synthesis in retinal explants indicate two mechanisms of apoptosis. One mechanism depends on the synthesis of positive modulators ('killer proteins'), while a distinct, latent mechanism appears to be continuously blocked by negative modulators. Extracellular modulators of apoptosis include the neurotrophic factors NT-4 and BDNF, while glutamate may have either a positive or a negative modulatory action on apoptosis. Several protein kinases selectively modulate apoptosis in distinct retinal layers. Calcium and nitric oxide were also shown to affect apoptosis in the developing retinal tissue. The protein c-Jun was found associated with apoptosis in various circumstances, while p53 seems to be selectively expressed in some instances of apoptosis. The results indicate that the sensitivity of each retinal cell to apoptosis is controlled by multiple, interactive, cell type- and context-specific mechanisms. Apoptosis in the retina depends on a critical interplay of extracellular signals delivered through neurotrophic factors, neurotransmitters and neuromodulators, several signal transduction pathways, and the expression of a variety of genes. PMID- 9394493 TI - The role of nitric oxide (NO) in control of hypothalamic-pituitary function. AB - Neurons containing neural nitric oxide synthase (nNOS) are found in various locations in the hypothalamus and, in particular, in the paraventricular and supraoptic nuclei with axons which project to the median eminence and extend into the neural lobe where the highest concentrations of NOS are found in the rat. Furthermore, nNOS is also located in folliculostellate cells and LH gonadotropes in the anterior pituitary gland. To define the role of NO in the release of hypothalamic peptides and pituitary hormones, we injected an inhibitor of NOS, Ng monomethyl-L-arginine (NMMA) or a releasor of NO, nitroprusside (NP) into the third ventricle (3V) of conscious castrate rats and determined the effect on the release of various pituitary hormones. In vitro, we incubated medial basal hypothalamic (MBH) fragments and studied inhibitors of NO synthase and also releasors of NO. The results indicate that NOergic neurons play an important role in stimulating the release of corticotrophin-releasing hormone (CRH), luteinizing hormone releasing-hormone (LHRH), prolactin-RH's, particularly oxytocin, growth hormone-RH (GHRH) and somatostatin, but not FSH-releasing factor from the hypothalamus. NO stimulates the release of LHRH, which induces sexual behavior, and causes release of LH from the pituitary gland. The intrahypothalamic pathway by which NO controls LHRH release is as follows: glutamergic neurons synapse with noradrenergic terminals in the MBH which release nonepinephrine (NE) that acts on alpha 1 receptors on the NOergic neuron to increase intracellular free Ca++ which combines with calmodulin to activate NOS. The NOS diffuses to the LHRH terminal and activates guanylate cyclase (GC), cyclooxygenase and lipoxygenase causing release of LHRH via release of cyclic GMP, PGE2 and leukotrienes, respectively. Alcohol and cytokines can block LHRH release by blocking the activation of cyclooxygenase and lipoxygenase without interfering with the activation of GC. GABA also blocks the response of the LHRH neurons to NO and recent experiments indicate that granulocyte macrophage colony-stimulating factor (GMCSF) blocks the response of the LHRH neuron to NP by activation of GABA neurons since the blockade can be reversed by the competitive inhibitor of GABAa receptors, bicuculine. PMID- 9394494 TI - Patterns of nitric oxide synthase expression in the developing superior colliculus. AB - Nitric oxide (NO) is synthesized in cells of both the central and peripheral nervous system and has been implicated in several forms of synaptic plasticity. The enzyme that produces NO, nitric oxide synthase (NOS), can be visualized in the brain by the reduced nicotinamide adenine dinucleotide phosphate diaphorase histochemistry technique (NADPH-d). We have used NADPH-d activity to detect the presence of NOS-positive cells in the developing rat superior colliculus. Our results showed that NOS is present in cells and neuropil in the developing and adult rat superior colliculus. The first NOS-positive cells appeared at postnatal day 7 and were weakly stained. The number and intensity of the NOS-positive cells increased progressively during the following days reaching a maximum at postnatal day 15. By the end of the third postnatal week, both the number and intensity of stained cells showed an adult-like pattern. The NOS-positive cells showed a Golgi like morphology and we have found that all cell types present in the superior colliculus express the enzyme. The expression of NOS by tectal cells parallels the functional development of the retino-collicular and cortico-tectal projections and suggest that nitric oxide synthase-positive cells might be involved in this process. In this review we highlighted some of the recent descriptions of the expression of NOS in the mammalian visual system with emphasis in the superior colliculus and correlate these findings with several developmental events taking place in this structure. PMID- 9394495 TI - Effects [correction of Effecs] of the thyroid hormone (T3) on astrocytes. AB - Thyroid hormones have profound effects on growth and development. In the brain L 3,5,3'-triiodothyronine (T3), the bioactive hormone, is involved with the harmonious development acting in neuronal and glial cell differentiation. T3 acts on the cells by interacting with nuclear receptors that can regulate the expression of several genes. Astrocytes also show receptors to the hormone. We reported herein data on the effects of T3 on astrocytes. We have verified that T3 has a morphological effect on cultured cortical astrocytes with rearrangement of GFAP filaments, and induces proliferation in the cultured cerebellar astrocytes of newborn rats. We discuss here the effects of T3 on astrocytes, considering the possibility that thyroid hormone prepares the astrocytes to interact with neurons. PMID- 9394497 TI - Development of nitric oxide synthase in the avian retina. AB - Nitric oxide is an important intercellular messenger in the central nervous system. Our previous work showed the presence of NADPH-diaphorase activity, that partially corresponded to nitric oxide synthase, in the chick embryo retina. In the present study, we have demonstrated the presence of nitric oxide synthase in the chick retina measuring the conversion of 3(H)arginine to 3(H)citrulline. We found that the enzyme is dependent on the presence of calcium, calmodulin and NADPH and is inhibited by the arginine analog L-NG-nitroarginine. The enzyme activity was higher at 8-day-old embryonic retinas, decreased at 13-14 days and attained minimal levels at 15 days up to the post-hatching period. Glutamate stimulated nitric oxide synthase activity approximately 4 fold, an effect that was blocked by the NMDA antagonist MK-801. The results indicate that the glutamate/nitric oxide system has important functions during retinal development. PMID- 9394496 TI - Neurotransmitter release in aggregate cultures of chick embryo retina cells. AB - The study of neurotransmitter release using aggregate cell cultures has been limited by the fact that cell aggregates are obtained only when cells are maintained in suspension with rotatory agitation. In this report we describe a simple and easy method that uses aggregate cell cultures to study the release of neurotransmitters. The results demonstrate that this relatively simple technique can be of great value to address the problem of neurotransmitter release and study the mechanisms of action of natural and synthetic compounds on the differentiation of functional synapses in the CNS. PMID- 9394498 TI - Monoamines and acetylcholine in primate cerebral cortex: what anatomy tells us about function. AB - In primates, cholinergic and monoaminergic axons that innervate the cerebral cortex originate almost exclusively from subcortical nuclei in the brainstem and basal forebrain. These projections are thought to modulate cortical activity during arousal, attention and memory formation. Physiological and anatomical evaluations of these ascending projections suggest that they have overlapping but somewhat distinctive synaptic targets in the cortex. This review compares the anatomical organization of acetylcholine-, dopamine-, norepinephrine-, and serotonin-containing axon systems in the monkey and human cerebral cortex. Analysis of the distributions of axons, receptors, and synapses suggests that each system is likely to have a differential role in modulating cortical function. PMID- 9394499 TI - Release and uptake of glutamate as related to excitotoxicity. AB - It has been established that neurons exposed to high concentrations of glutamate or other excitatory amino acids degenerate and die. Neuronal damage appears to be due to the activation of different types of glutamate receptors, among which the ionotropic N-methyl-D-aspartate (NMDA) type seems particularly involved, since its channel is permeable to Ca2+ and an increase in the cytoplasmic concentration of this cation promotes a chain of events leading to cell death. The mechanism of such glutamate receptor-mediated neurodegeneration has been defined as excitotoxicity, and several pieces of evidence suggest that this mechanism might contribute to the neuronal death associated with certain neurological disorders, such as ischemia, cerebral trauma and some chronic neurodegenerative diseases. A relevant question is whether the origin of endogenous extracellular glutamate is important for the induction of excitotoxicity. An excess of glutamate release, or a deficiency in its clearance from the synaptic cleft, which depends mainly on its transport by high affinity carriers, are potential sources for the accumulation of extracellular glutamate. In the present article some experimental results from our laboratory, aimed at obtaining information on this question, are reviewed. These experiments include the use of 4-aminopyridine, a convulsant drug that enhances the release of glutamate, and of some inhibitors of glutamate transport, in vivo and in neuronal cell cultures. The results obtained indicate that an increase of endogenous extracellular glutamate due to these procedures is not sufficient to induce neuronal death, at least under the experimental conditions used. PMID- 9394500 TI - Subcellular localisation of neurotrophins and neurotrophin receptors: implications for synaptic plasticity. AB - A growing body of recent evidence indicate that neurotrophins can act as mediators of neuronal plasticity. In the context of a more detailed, comprehensive understanding of the function of neurotrophins it is essential to characterize where neurotrophins are synthesised and stored and from where they are released. Here we present evidence that the mRNAs for NGF, trkB and BDNF but not trkA are localised in the dendrites of rat neurons, thus implying that neurotrophins and their receptors can be synthesised at locations close to their sites of function, with particular regard to the dendritic synapses. The significance of this finding and its possible implications for synaptic plasticity are discussed within the theoretical framework of the synapse-specific control of individual synapses of a given neuron. PMID- 9394501 TI - The opossum photoreceptors--a model for evolutionary trends in early mammalian retina. AB - The topography and spectral characteristics of mammalian photoreceptors correlate with both, the present ecological demands and the evolutionary history. The South American Opossum is a marsupial mammal with unspecialized habitus and crepuscular lifestyle. A sparse population of cones (max. = 3000/mm2) can be differentiated into four subtypes by morphological, topographical and immunocytochemical criteria. In spite of this unusual diversity the cone types can be split into two functional groups: The population of single cones labeled by antibody OS-2 for short wavelength sensitive pigments was ubiquitous but at very low densities (200/mm2). The single cones labeled by antibody (COS-1) against long wavelength sensitive pigments constitute the dominant population in the area centralis (2300/mm2). These two single cone types correlate with the pair typically present in placental mammals. Discrimination of spatial and color contrast may be provided by this "modern" set. The COS-1 labeled double and single cones bearing an oil droplet, display a different pattern by being restricted to the inferior (non-tapetal) half of the retina (max = 800/mm2). This additional set of cones with oil droplets and long wavelength pigments is a conservative feature of the opossum retina and other marsupials. As an accessory cone system it is possibly providing enhanced sensitivity at mesopic conditions. During the early evolution of nocturnal mammals with its prominent expansion of rod vision these cone types were conserved but then were lost in placental mammals. Thus the unique features of mammalian retinas are the result of two evolutionary steps: first a reduction of cone based vision, followed by a secondary differentiation of photopic vision and behaviour relying on the remaining set of cones. PMID- 9394502 TI - Intrinsic projections of Cebus-monkey area 17: cell morphology and axon terminals. AB - Metric features of the axon terminals and cell morphology of intrinsic projections of area 17 were studied in the Cebus apella. Anterograde and retrograde labeled cell appendages were obtained using saturated pellets and iontophoretic injections of biocytin in the operculum. Details of the histological and histochemical procedures have been described elsewhere (Amorim and Picanco-Diniz, 1996). We distinguished three labeled cell types: pyramidal, star pyramidal and stellate cells and three distinct morphologies of axon terminals were found: Ia, Ib and II, located at supragranular layers. Axon terminals of the group I innervate larger extent of striate cortex through longer intermediate segments, and acute branching angles compared to group II. Group II present on average similar characteristics of the smooth neurons axon terminals. The results taken together with the occurrence of only two types of synapses (I and II) from Gray's ultrastructural studies, seem to give an additional support to extend to the Cebus apella the major subdivision of neocortical neuronal morphology that classified them as smooth and spine neurons. PMID- 9394503 TI - ANP as a neuroendocrine modulator of body fluid homeostasis. AB - The role played by the central nervous system (CNS) in the control of body fluid homeostasis has been demonstrated by several authors. The AV3V plays a key role in central control of sodium excretion since its cholinergic, adrenergic, angiotensinergic and osmotic stimulation enhances and its destruction blocks sodium excretion in rats and goats. Cholinergic stimulation of the AV3V induced an increase in plasma ANP as well as a marked elevation in content of the peptide in medial basal hypothalamus, neuro and adenohypophysis. On the other hand, a decline in plasma ANP after AV3V lesions was accompanied by dramatic declines in content of ANP in these same structures. Our previous work has also indicated the essential role of the AV3V region and its ANPergic neurons in the control of ANP release in response to volume expansion (BVE) and indicated that alpha-adrenergic and muscarinic receptors are critical in mediating these responses. Lesions of the AV3V region, or of the median eminence or posterior lobe of pituitary gland blocked the increase in plasma ANP concentration in response to BVE. That this effect is related to blockage of the activity of the brain ANPergic neurons is supported by findings in sheep and in rats that the injection of the antiserum directed against ANP into the AV3V region at least partially blocked the BVE induced release of ANP. We and others have also previously shown that denervation of baroreceptors inhibits ANP release induced by BVE. Activation of the ANP neurons also cause release of ANP from the anterior and neural lobe of pituitary gland. ANP neurons may activate oxytocinergic neurons in the supraoptic and paraventricular, which projects to neural lobe. Oxytocin would circulate to the atria and may directly activate release of ANP from the atrial myocytes, since i.v. or i.p. injection of oxytocin increases sodium excretion as well as elevates plasma ANP. Oxytocin is present in the neural lobe in large quantity, which could reach the atria myocytes in high concentration and release ANP that circulate to the kidneys and evokes natriuresis to return circulating blood volume to normal. PMID- 9394504 TI - Preoptic-periventricular tissue (AV3V): central cholinergic-induced hydromineral and cardiovascular responses, and salt intake. AB - The periventricular tissue of the anterior ventral portion of the third ventricle (AV3V) is an important area for the control of hydromineral balance and of cardiovascular function. The present work discusses the importance of the integrity of the AV3V for multiple responses to central cholinergic activation (water intake, hypertension, natriuresis, salivation) and for the control of salt intake. PMID- 9394505 TI - The primate frontal eye field and the generation of saccadic eye movements: comparison of lesion and acute inactivation/activation studies. AB - The frontal eye field (FEF) of monkeys has been repeatedly implicated in the generation of saccadic eye movements by various experimental approaches. Electrical stimulation of most of the FEF produces saccadic eye movements, many cells have activities related to saccades, and it has anatomical connections with many other oculomotor areas. Surprisingly, complete lesions of the FEF have remarkably little effect on oculomotor behavior. Only when more cognitive aspects are tested is a deficit clearly detected. In contrast, acute inactivation of the FEF of monkeys with the GABA agonist muscimol produced much more severe oculomotor impairment. This difference is probably due to the acute nature of the muscimol effect, which does not allow time for reorganization of the control of eye movements before testing begins. In addition, acute activation of the FEF with the GABA antagonist bicuculline caused the monkey to make irrepressible saccades of the same dimensions as those electrically elicited at the site. These experiments further confirm the strong involvement of the FEF in the control of saccadic eye movements and fixation. PMID- 9394506 TI - Responses of cells in the superior colliculus during performance of a spatial attention task in the macaque. AB - Previous studies have reported that superficial layer cells in the superior colliculus (SC) give an enhanced response to a stimulus when it is the target for an eye movement. However, in a peripheral detection paradigm, no such enhancement was found when a stimulus was attended, in the absence of an eye movement. Inasmuch as behavioral studies have found attention deficits in the absence of eye movements following SC lesions or deactivation, we investigated this issue in a paradigm that is very sensitive to effects of attention. In a matching-to sample paradigm, a sample stimulus was presented at one location followed by a brief test stimulus at that (relevant) location and a distracter at another (irrelevant) location. While maintaining fixation, the monkey indicated whether the sample and the test stimulus matched, ignoring the distracter. The relevant and irrelevant locations were switched from trial to trial. SC cells in the superficial layers tended to give enhanced responses when the attended test stimulus was inside the receptive field compared to when the (physically identical) distracter was inside the field. We found that responses to attended targets in the receptive field were larger than to physically identical, but ignored, distracter stimuli. These effects were found only in an "automatic" attentional cueing paradigm, in which a peripheral stimulus explicitly cued the animal as to the relevant location in the receptive field. No attentional effects were found in a "central" or "cognitive" cueing paradigm, in which the monkey had to learn the relevant location in a given block of trials. The larger responses to attended targets in the automatic cueing paradigm appeared to be due to a sustained elevation of cells' baseline activity when attention was directed to the receptive field, as well as a transient enhancement of the target response. Thus, responses of SC cells appear to be modulated by directed attention, even in absence of eye movements, probably reflecting the properties of cortical cells projecting to the SC. PMID- 9394507 TI - Interaction between facilitatory and inhibitory effects due to voluntary and automatic covert orienting of attention. AB - Covert orienting of attention to one spatial location improves the processing of signals occurring at this location at the expenses of the processing of signals occurring at other spatial positions. According to the premotor theory of visual attention, the voluntary orienting of attention to a peripheral position corresponds to the programming of a saccadic eye movement towards this position. A similar mechanism has been proposed to explain the inhibitory effects elicited by a non-informative peripheral cue. This review discusses some neural mechanisms involved in the facilitatory and inhibitory effects due to covert orienting of attention. PMID- 9394508 TI - Malnutrition and brain function: experimental studies using the phenomenon of cortical spreading depression. AB - Depending on its intensity and duration, nutritional deficiency can disrupt the structure and function of the nervous system of humans and other mammals, with consequences more or less devastating for the whole organism, particularly in the early postnatal life, when body growth is very rapid and the need for proteins, calories and other nutrients is greatest. In this review, electrophysiological data are presented regarding the use of the phenomenon of cortical spreading depression (CSD) to study effects of malnutrition on the brain. Several conditions of clinical importance and that are known to alter brain function are shown also to influence CSD features in experimental animals. Some of these conditions, (e.g., pharmacological manipulation of neurotransmitter systems, dietary treatment with Lithium, acute hyperglycemia, hypothyroidism, aging and environmental stimulation) decrease CSD susceptibility, while other conditions increase it, as, for example, systemic reduction of extracellular chloride levels, deprivation of REM-sleep, acute hypoglycemia, treatment with diazepam, consumption of ethanol and malnutrition. Particular emphasis is laid on the effect of early environmental enrichment on CSD in normal and malnourished animals. Our results suggest that such effect is more evident in the malnourished brain, as compared to the well-nourished one. The data also show that malnutrition alters the brain responsivity to some CSD-facilitatory or inhibitory agents. The underlying mechanisms to explain the observed effects are discussed. PMID- 9394509 TI - Comparative neurobiology of the optokinetic reflex in mammals. AB - Mammals have developed almost perfect functional adaptations of their eyes, central visual system and oculomotor system for high resolution and stable vision. This performance is subserved by stabilizing reflexes like the optokinetic and the vestibular ocular reflex. Visual information about retinal image slip is relayed through the nucleus of the optic tract in the pretectum and the terminal nuclei of the accessory optic system to the visual motor system. Retinal and more importantly cortical information is integrated in these nuclei to optimize visual control of gaze stabilization during ego-motion. PMID- 9394510 TI - What comparative studies of neocortex tell us about the human brain. AB - There are several ways in which comparative studies of brain organization and function can be informative in attempts to understand the human brain. Often investigators study a favorable and sometimes specialized species in order to reveal features that may reflect general or widespread principles. The example used here is that the cortical representation of the unique and highly specialized receptor sheet of the nose of the star-nosed mole provides further evidence that the receptor sheet instructs the development of cortex. Comparative studies are also used to reconstruct the evolution of brain systems. As an example, comparative studies suggest that the visual area MT in the upper temporal lobe of primates evolved from a visual area along the border of V2. A third and very important use of comparative studies is to provide another level of evaluation of theories developed from a particular species. Since the brains of different mammals are modifications of an ancestral plan, conclusions about brain organization in any given species should be consistent with those for other species, within the framework of evolutionary change. When current proposals for how visual cortex is organized in rats and humans are considered from a comparative point of view, key concepts are clearly challenged. PMID- 9394511 TI - Parallel pathways in the retina of Old and New World primates. AB - Old-world simians are all trichromats, but in most new-world primates there is a polymorphism; males are dichromats but most females are trichromats. In the old world simian, luminance and red-green chromatic channels defined by psychophysical experiments have as a basis parasol ganglion cells of the magnocellular (MC) pathway and midget ganglion cells of the parvocellular (PC) pathway respectively. Small bistratified ganglion cells provide a basis for a blue-yellow chromatic channel, which should probably be considered a separate entity. In both dichromatic and trichromatic new-world animals, the MC pathway and the small bistratified, blue-yellow system seem anatomically and physiologically similar to those in their old-world relatives. The midget ganglion cells of the parvocellular pathway in trichromats are anatomically and physiologically similar to the old-world pattern. In dichromatic animals, they are anatomically similar and physiologically resemble those of trichromatic animals, except for the lack of chromatic opponency. We conclude that these three systems may from a basic pattern for the visual pathway of primates. However, the results from dichromats indicate that the evolution of trichromacy may be found to be more complex than presently supposed. PMID- 9394512 TI - Some aspects of the pineal gland function. AB - Rhythmic production of melatonin by the mammalian pineal gland occurs in response to noradrenergic stimulation which produces a cascade of biochemical events within the pinealocyte. Melatonin is involved in a wide range of physiological processes, including seasonal reproduction and diurnal activity rhythms. It was recently discovered that melatonin is a very potent hydroxyl radical scavenger, reinforcing its relationship with the aging process and the immune system. PMID- 9394513 TI - Joint entropy loci of M and P cells: a hypothesis for parallel processing in the primate visual system. AB - The M and P neurons connect the retinal inner plexiform layer to the magnicellular and parvicellular layers of the lateral geniculate nucleus and to layer 4 of the primary visual cortex. The M and P pathways transmit visual signals with different six-dimensional joint entropy for space, spatial frequency, time and temporal frequency. Beyond V1 layer 4, M and P influences mix in information processing streams which connect different visual cortical areas. Some visual cortical cells perform better than it is possible for linear filters, regarding to simultaneous spatial and spatial frequency localization of their response. This can be explained by nonlinear combination of M and P input on such cells. PMID- 9394514 TI - On the purinergic regulation of hormonal secretion from the anterior pituitary gland. AB - Purinergic regulation of hormonal secretion from the anterior pituitary may be characterized by effects with biphasic secretory response. This response may be started by activation of different subtypes of membrane prurinergic receptors (A1 and/or A2). A putative autocrine mechanism has been proposed to explain the action of adenosine on pituitary hormonal secretion. This mechanism may be dependent on adenosine degradation by the enzyme adenosine deaminase into the extracellular space. The regulation of AMPc and calcium levels in cytoplasm may be part of putative intracellular mechanisms involved in purinergic action. Additionally, hypophysiotrophic effects induced by hypothalamic substances may be modulated by adenosine. The mechanisms involved in this modulatory effects, however, remain elusive. PMID- 9394515 TI - The nucleus of the optic tract of the opossum. AB - This paper reviews anatomical and electrophysiological data on the nucleus of the optic tract (NOT) of the opossum, a nucleus in the afferent branch of the horizontal optokinetic reflex. It is proposed that subcortical routes are essential for responses from the two eyes: a direct retinal projection from the contralateral eye and a commissural pathway between the two NOTs for the ipsolateral eye. In the latter case there's evidence that the commisural axons have a relay on inhibitory neurones. This circuit accounts for the differences in response pattern under monocular condition: temporo-nasal motion of the visual stimulus elicits excitation in the contralateral NOT, resulting in inhibition of the ipsolateral nucleus, while naso-temporal motion promotes inhibition in the contralateral nucleus, releasing the ipsolateral nucleus from the commissural input. PMID- 9394516 TI - The retinal ganglion cell classes of New World primates. AB - In the primate retina there are distinct ganglion cell classes, exhibiting particular morphologies and central projections, each responsible for conveying particular types of visual information to the brain. The chief retinal inputs to the cortex arise from specific ganglion cell classes, M-ganglion cells, responsible for carrying the luminance signal, and P-ganglion cells, that convey the red-green color opponent signal, as well as high contrast luminance signal. There are other ganglion cell classes, such as small-field bistratified cells, exhibiting dendrites that stratify at two different levels in the inner plexiform layer, which convey the blue-yellow color opponent signal. Most published data concerning primate retinal ganglion cell anatomy and physiology have been obtained from Old World species. Studies on New World monkeys have recently become of interest since they differ from the Old World monkeys with respect to the color vision inheritance pattern. On reviewing retinal ganglion cell layer organization in New World monkeys, it seems that there are more similarities than differences in relation to the Old World monkeys. Diurnal genera of New World monkeys exhibit a well-developed fovea centralis and ganglion cell density peak, as well as peripheral density values which are in the range reported for Old World monkeys and human. Moreover, all the major ganglion cell classes identified in Old World monkeys are also present in New World primates. Up to now, no obvious anatomical differences between dichromats and trichromats have been reported. The only genus that is significantly different from the others is the Aotus. It exhibits lower ganglion cell density in the central retina, and apparently lacks the small-field bistratified cells. PMID- 9394517 TI - Virulence parameters in the characterization of strains of Entamoeba histolytica. AB - Differences in virulence of strains of Entamoeba histolytica have long been detected by various experimental assays, both in vivo and in vitro. Discrepancies in the strains characterization have been arisen when different biological assays are compared. In order to evaluate different parameters of virulence in the strains characterization, five strains of E. histolytica, kept under axenic culture, were characterized in respect to their, capability to induce hamster liver abscess, erythrophagocytosis rate and cytopathic effect upon VERO cells. It was found significant correlation between in vitro biological assays, but not between in vivo and in vitro assays. Good correlation was found between cytopathic effect and the mean number of uptaken erythrocytes, but not with percentage of phagocytic amoebae, showing that great variability can be observed in the same assay, according to the variable chosen. It was not possible to correlate isoenzyme and restriction fragment pattern with virulence indexes since all studied strains presented pathogenic patterns. The discordant results observed in different virulence assays suggests that virulence itself may not the directly assessed. What is in fact assessed are different biological characteristics or functions of the parasite more than virulence itself. These characteristics or functions may be related or not with pathogenic mechanisms occurring in the development of invasive amoebic disease. PMID- 9394518 TI - Immune responses induced by a Leishmania (Leishmania) amazonensis recombinant antigen in mice and lymphocytes from vaccinated subjects. AB - In the search for Leishmania recombinant antigens that can be used as a vaccine against American Cutaneous Leishmaniasis, we identified a Leishmania (Leishmania) amazonensis recombinant protein of 33 kD (Larp33) which is recognized by antibodies and peripheral blood leukocytes (PBL) from subjects vaccinated with Leishvacin, Larp33 was expressed in Escherichia coli after cloning of a 2.2 kb Sau3 digested genomic fragment of L. (L.) amazonensis into the pDS56-6 His vector. Immunoblotting analysis indicated that Larp33 corresponds to an approximately 40-kD native protein expressed in promastigotes of L. (L.) amazonensis and L. (Viannia) braziliensis. Northern blots of total RNA also demonstrated that the gene coding for this protein is expressed in promastigotes of the major lineages of Leishmania causing American Cutaneous Leishmaniasis. Larp33 induced partial protection in susceptible mouse strains (BALB/c and C57BL/10) against L. (L.) amazonensis after vaccination using Bacille Calmette Guerin (BCG) as adjuvant. In vitro stimulation of splenocytes from BALB/c protected mice with Larp33 elicited the secretion of IL-2 and IFN-gamma, suggesting that a Th1 cell-mediated protective response is associated with the resistance observed in these mice. As revealed by its immunogenic and antigenic properties, this novel recombinant antigen is a suitable candidate to compose a vaccine against cutaneous leishmaniasis. PMID- 9394519 TI - Detection and identification of dengue virus isolates from Brazil by a simplified reverse transcription-polymerase chain reaction (RT-PCR) method. AB - We show here a simplified RT-PCR for identification of dengue virus types 1 and 2. Five dengue virus strains, isolated from Brazilian patients, and yellow fever vaccine 17DD as a negative control, were used in this study. C6/36 cells were infected and supernatants were collected after 7 days. The RT-PCR, done in a single reaction vessel, was carried out following a 1/10 dilution of virus in distilled water or in a detergent mixture containing Nonidet P40. The 50 microliters assay reaction mixture included 50 pmol of specific primers amplifying a 482 base pair sequence for dengue type 1 and 210 base pair sequence for dengue type 2. In other assays, we used dengue virus consensus primers having maximum sequence similarity to the four serotypes, amplifying a 511 base pair sequence. The reaction mixture also contained 0.1 mM of the four deoxynucleoside triphosphates, 7.5 U of reverse transcriptase, 1U of thermostable Taq DNA polymerase. The mixture was incubated for 5 minutes at 37 degrees C for reverse transcription followed by 30 cycles of two-step PCR amplification (92 degrees C for 60 seconds, 53 degrees C for 60 seconds) with slow temperature increment. The PCR products were subjected to 1.7% agarose gel electrophoresis and visualized by UV light after staining with ethidium bromide solution. Low virus titer around 10(3, 6) TCID50/ml was detected by RT-PCR for dengue type 1. Specific DNA amplification was observed with all the Brazilian dengue strains by using dengue virus consensus primers. As compared to other RT-PCRs, this assay is less laborious, done in a shorter time, and has reduced risk of contamination. PMID- 9394520 TI - In vivo and in vitro Plasmodium falciparum resistance to chloroquine, amodiaquine and quinine in the Brazilian Amazon. AB - In order to study the chemoresistance of Plasmodium falciparum to commonly used antimalarial drugs in Brazil the authors have studied ten patients with falciparum malaria, acquired in the Brazilian Amazon region. Patients were submitted to in vivo study of drug sensitivity, after chemotherapy with either 4 aminoquinolines (chloroquine or amodiaquine) or quinine. Adequate drug absorption was confirmed by standard urine excretion tests for antimalarials. Eight patients could be followed up to 28 days. Among these in vivo resistance (R I and R II responses) was seen in all patients who received 4-amino-quinolines. One patient treated with quinine exhibited a R III response. Peripheral blood samples of the same patients were submitted to in vitro microtests for sensitivity to antimalarials. Out of nine successful tests, resistance to chloroquine and amodiaquine was found in 100% and resistance to quinine in 11.11% of isolates. Probit analysis of log dose-response was used to determine effective concentrations EC50, EC90 and EC99 to the studied drugs. Good correlation between in vivo and in vitro results was seen in six patients. The results emphasize high levels of P. falciparum resistance to 4-aminoquinolines and suggest an increase in resistance to quinine in the Brazilian Amazon region, reinforcing the need for continuous monitoring of drug sensitivity to adequate chemotherapy according to the most efficacious drug regimens. PMID- 9394521 TI - Water-contact patterns and risk factors for Schistosoma mansoni infection in a rural village of northeast Brazil. AB - Schistosomiasis mansoni in the Serrano village, municipality of Cururupu, state of Maranhao, Brazil, is a widely spread disease. The PECE (Program for the Control of Schistosomiasis), undertaken since 1979 has reduced the prevalence of S. mansoni infection and the hepatosplenic form of the disease. Nevertheless piped water is available in 84% of the households, prevalence remains above 20%. In order to identify other risk factors responsible for the persistence of high prevalence levels, a cross-sectional survey was carried out in a systematic sample of 294 people of varying ages. Socioeconomic, environmental and demographic variables, and water contact patterns were investigated. Fecal samples were collected and analyzed by the Kato-Katz technique. Prevalence of S. mansoni infection was 24.1%, higher among males (35.5%) and between 10-19 years of age (36.6%). The risk factors identified in the univariable analysis were water contacts for vegetable extraction (Risk Ratio--RR = 2.92), crossing streams (RR = 2.55), bathing (RR = 2.35), fishing (RR = 2.19), hunting (RR = 2.17), cattle breeding (RR = 2.04), manioc culture (RR = 1.90) and leisure (RR = 1.56). After controlling for confounding variables by proportional hazards model the risks remained higher for males, vegetable extraction, bathing in rivers and water contact in rivers or in periodically inundated parts of riverine woodland (swamplands). PMID- 9394522 TI - Evaluation of the schistosomicidal efficacy of liposome-entrapped oxamniquine. AB - Oxamniquine (OXA) was successfully encapsulated in small unilamellar vesicles using a pH gradient method. This procedure led to a high drug encapsulation efficiency (> 85%) at a drug to lipid molar ratio of 1/10. Moreover, these liposomes were found to retain encapsulated OXA efficiently under dialysis conditions at 37 degrees C. Liposome-entrapped OXA (LOXA), OXA, and empty liposomes were tested against Schistosoma mansoni in a murine model. LOXA produced a significant reduction of the worm burden compared to the other preparations, when inoculated by subcutaneous route (s.c.) with 10 mg OXA/kg animal one day before the infection, and 3, 7, and 14 days after. However, LOXA was not effective when given 7 days before, or 35 days after infections. OXA, in the free form, was effective in relation to the untreated group, only when administered 3 days after the infection. Maximum effect of LOXA, with 97% reduction of the parasite number, was observed when the preparation was given s.c. one day before the infection. On the other hand, LOXA inoculated intraperitoneally one day before the infection didn't show any reduction of the parasite count. It can be concluded that LOXA is more effective than OXA for the treatment of experimental schistosomiasis, particularly when administered subcutaneously at a time close to the infection. PMID- 9394523 TI - Resistance to oxamniquine of a Schistosoma mansoni strain isolated from patient submitted to repeated treatments. AB - A strain of Schistosoma mansoni (R1) was isolated from patient previously submitted to four treatments with oxamniquine, and to another one with praziquantel. The results obtained with chemotherapeutic test, by using oxamniquine in mice infected with the strains R1 and LE (standard), showed an evident resistance to the drug in worms of the strain R1. Thus, at the dose of 250 mg/kg oxamniquine, all mice (17) infected with the LE strain did not show surviving worms, whereas 12 out of 17 mice infected with the R1 strain presented surviving worms. At the dose of 200 mg/kg, the LE strain showed recovery rates of 1.06% and 20.58%, whereas the R1 strain presented 18.57% and 61.14%, for male and female worms, respectively. At the dose of 100 mg/kg, the recovery of male worms was 2.6% for the LE strain, and 29.9% for the R1 strain. At the same dose, the recovery of females did not show statistically significant differences between the two strains (LE = 76.38%, R1 = 79.12%). Praziquantel showed similar antischistosomal activity against both studied strains, when administered at the dose of 500 mg/kg. PMID- 9394524 TI - Human infection with Trypanosoma cruzi in Nasca, Peru: a seroepidemiological survey. AB - We estimated the proportion of seropositivity for infection with Trypanosoma cruzi (Chagas' disease) in a sample of the rural population of the Province of Nasca, Department of Ica, southwestern Peru. Although Triatoma infestans, the only vector species identified in the Department of Ica, is often found in domestic environments, data of the extent of human infection with T. cruzi are scant. This study comprised 446 houses, known to be infested with triatomines, distributed in 19 rural localities. While visiting those houses we collected filter paper bloodspots from 864 occupants (of both sexes, aged one year or over). By means of the indirect fluorescent antibody test (IFAT), we detected anti-T. cruzi IgG antibodies in samples from 178 individuals (20.6%). Seropositivity was significantly more frequent in females (23.8%) than in males (17.5%). Among the 410 individuals in the 1- to 10-year-old age group (47.5% of the population sample), 85 (20.7%) were found seropositive, which is indicative of an early acquisition of the infection. Within this group no significant differences in seropositivity were associated with sex. PMID- 9394525 TI - Epidemiological and clinical aspects of snakebite in Belo Horizonte, southeast Brazil. AB - Epidemiologic and clinical aspects of 310 hospitalized snakebite patients and 310 matched controls were described, over a seven years period, from an emergency hospital in Belo Horizontal, Southeast Brazil. The diagnosis was based upon clinical picture or actual snake identification. Fifty six percent of victims were bitten by the snakes of genus Bothrops, 32.0% by Crotalus, 1.0% by Lachesis and 10.0% undetermined. During the study period, stable number of cases and marked seasonal variation were noted. In comparing cases of snakebite and controls, those from a rural area or who were involved in agricultural labor activity were identified as a high risk group, with an odds ratio (OR) of 14.7 and 6.7, respectively, in favor of being bitten. Upon treatment, snakebite patients were 13.5 times more likely to have had early anaphylactic reactions than their controls, with a higher association in the age group > or = 20 years (OR = 30.3). Increased risks were also detected for pyrexia (OR = 11.7), with a marked association in the group under 19 years old (OR = 16.6). Severe cases of snakebite are an important treatable cause of morbidity in Brazil but therapy may be potentially life threatening. The higher case-fatality ratio encountered, compared to national statistics may be due the representativeness of the more severe cases who sought hospitalization. Preventing snakebite and early referral of those who are bitten is proposed. PMID- 9394526 TI - Effect of beta-propiolactone treatment on the complement activation mediated by equine antisera. AB - Reduction of complement activation through an alteration of the Fc fragment of immunoglobulins by beta-propiolactone treatment was carried out in equine antisera raised against rabies virus, Bothrops venoms and diphtherial toxin. Results were evaluated by means of an anaphylactic test performed on guinea-pigs, and compared to the ones obtained with the same sera purified by saline precipitation (ammonium sulfate), followed or not by enzymatic digestion with pepsin. Protein purity levels for antibothropic serum were 184.5 mg/g and 488.5 mg/g in beta-propiolactone treated and pepsin-digested sera, respectively. The recovery of specific activity was 100% and 62.5% when using antibothropic serum treated by beta-propiolactone and pepsin digestion, respectively. The antidiphtherial and anti-rabies sera treated with beta-propiolactone and pepsin presented protein purity levels of 5,698 and 7,179 Lf/g, 16,233 and 6,784 IU/g, respectively. The recovery of specific activity for these antisera were 88.8%, 77.7%, 100% and 36.5%, respectively. beta-propiolactone treatment induced a reduction in complement activation, tested "in vivo", without significant loss of biological activity. This treatment can be used in the preparation of heterologous immunoglobulins for human use. PMID- 9394527 TI - Bertiellosis in man: a review of cases. AB - The presence of Bertiella mucronata and Bertiella studeri (Cestoda: Anoplocephalidae) in humans is reviewed, and international infection rates and a bibliography included. Taxonomic, biological, epidemiological, pathological, diagnostic, control, prevention and therapeutic aspects of the zoonosis are analyzed, and the increase in zoonotic potentiality of the parasitosis is discussed. PMID- 9394528 TI - A focus of favus due to Trichophyton schoenleinii in Rio Grande do Sul, Brasil. PMID- 9394529 TI - Description of the egg of Anopheles (Anopheles) intermedius (Peryassu, 1908) (Diptera: Culicidae) by scanning electron microscopy. AB - The egg of Anopheles (Anopheles) intermedius (Peryassu, 1908) is described and illustrated with scanning electron micrographs. Literature data on An. (Ano.) maculipes (Theobald, 1903) is provided. PMID- 9394530 TI - Immunostimulation as adjuvant for the chemotherapy of experimental schistosomiasis. AB - Immunosuppressed animals respond poorly to schistosomal chemotherapy and a proper response can be restored by the administration of immune serum. Present study attempts to search whether immunological stimulation would increase drug effectiveness. Swiss mice infected with 50 S. mansoni cercariae were later treated with complete Freund's adjuvant. Treatment with oxamniquine was made with 100 mg/kg.b.w., 25 mg/kg.b.w. and 50 mg/kg.b.w., the last two doses representing a fourth and a half of the recommended curative dose. Appropriate controls for the drug, the adjuvant and the infection were also studied. The serum-level of anti-S. mansoni antibodies (ELISA) and recovery of worms by perfusion of the portal vein system were the evaluated parameters. Statistical analysis of the results failed to reveal significant differences in worm recovery between adjuvant-stimulated animals treated with oxamniquine and any of the treated controls receiving the same amount of the drug. Although total lack of immunity interferes with curative treatment the usual immune response seems to be sufficient to allow for curative drug action in schistosomiasis and thus apparently does not need to be artificially stimulated. PMID- 9394531 TI - Immunogenicity of low-dose intramuscular and intradermal vaccination with recombinant hepatitis B vaccine. AB - The study is a randomized trial using recombinant DNA vaccine to determine whether an intramuscular 10 micrograms dose or intradermal 2 micrograms induces satisfactory anti-HBs levels compared to the standard dose of intramuscular 20 micrograms. Participants were 359 healthy medical and nurse students randomly allocated to one of the three groups: Group I-IM 20 micrograms; Group II-IM 10 micrograms; Group III-ID 2 micrograms at 0, 1 and 6 months. Anti-HBs titres were measured after complete vaccine schedule by ELISA/Pasteur. Baseline variables were similar among groups and side effects were mild after any dose. Vaccines in the IM-10 micrograms group had seroconversion rate and geometric mean titre (GMT 2344 IU L-1), not significant different from the IM-20 micrograms group (GMT 4570 IU L-1). On the contrary, 21.4% of the ID-2 micrograms recipients mount antibody concentration below 10 IU L-1 and GMT of 91 IU L-1, a statistically significant difference compared with the standard schedule IM-20 micrograms (p < 0.001). A three dose regimen of half dose IM could be considered an appropriate schedule to prevent hepatitis B in young health adults which is of relevance to the expansion of hepatitis B vaccine programme. PMID- 9394532 TI - Evaluation of a rapid screening assay for bacterial identification (Dot-ELISA) in fecal samples from children. AB - With the objective of standardizing a Dot Enzyme-Linked Immunosorbent Assay (Dot ELISA) to detect antigens of fecal bacterial enteropathogens, 250 children, aged under 36 months and of both sexes, were studied; of which 162 had acute gastroenteritis. The efficacy of a rapid screening assay for bacterial enteropathogens (enteropathogenic Escherichia coli "EPEC", enteroinvasive Escherichia coli "EIEC", Salmonella spp. and Shigella spp.) was evaluated. The fecal samples were also submitted to a traditional method of stool culture for comparison. The concordance index between the two techniques, calculated using the Kappa (k) index for the above mentioned bacterial strains was 0.8859, 0.9055, 0.7932 and 0.7829 respectively. These values express an almost perfect degree of concordance for the first two and substantial concordance for the latter two, thus enabling this technique to be applied in the early diagnosis of diarrhea in infants. With a view to increasing the sensitivity and specificity of this immunological test, a study was made of the antigenic preparations obtained from two types of treatment: 1) deproteinization by heating; 2) precipitation and concentration of the lipopolysaccharide antigen (LPS) using an ethanol-acetone solution, which was then heated in the presence of sodium EDTA. PMID- 9394533 TI - Hemagglutination test for the diagnosis of human neurocysticercosis: development of a stable reagent using homologous and heterologous antigens. AB - A hemagglutination (HA) test was standardized using formalin- and tannin-treated gander red blood cells sensitized with a total salt extract of C. cellulosae (HA Cc) and an antigenic extract of Cysticercus longicollis (HA-Cl) vesicular fluid. A total of 61 cerebrospinal fluid (CSF) samples were assayed, 41 from patients with neurocysticercosis and 20 from a control group which were, respectively, reactive and non-reactive to ELISA using C. cellulosae. The CSF samples from the control group did not react and 35 (85.4%) and 34 (82.9%) CSF samples from patients were reactive to the HA-Cc and HA-Cl tests, respectively. The reagents ready for use were stable up to 6 months when stored at 4 degrees C in 50% glycerol. The present results confirm that the reagent using Cysticercus longicollis stabilized with glycerol can be used as an alternative in the immunological diagnosis of neurocysticercosis. PMID- 9394534 TI - Reactivity of anti-gp43 antibodies from Paracoccidioides brasiliensis antiserum with extracts from cutaneous lesions of Lobo's disease. Preliminary note. AB - We demonstrated through several immunochemical tests the presence of gp43 from P. brasiliensis in extracts of cutaneous lesions from Jorge Lobo's disease. This glicoprotein is one of the immunodominant antigens in this species, and is used to identify it. The demonstration of gp43 tissues infected by the agent of Jorge Lobo's disease is an additional evidence for classifying it in the genera Paracoccidioides, species loboi. PMID- 9394535 TI - Preliminary report of the use on adults of a recombinant yeast-derived hepatitis B vaccine manufactured by Instituto Butantan. AB - Three 10 micrograms of the recombinant hepatitis B vaccine, manufactured by Instituto Butantan by original technology, were administered in an adult population, mean age 30 years old, following the 0, 1 and 6 months schedule immunization. The clinical trial was considered satisfactory in terms of immunogenicity (anti-HBs titers between 17.5-29500 IU/ l, seroconversion 95.3%) and reactogenicity (no incapacitating side effects). PMID- 9394536 TI - Subcutaneous phaeohyphomycosis caused by Phoma cava. Report of a case and review of the literature. AB - We report a case of subcutaneous phaeohyphomycosis observed in a male patient presenting pulmonary sarcoidosis and submitted to corticosteroid treatment. He presented nodular erythematous-violaceous skin lesions in the dorsum of the right hand. Histopathological examination of the biopsied lesion revealed dematiaceous hyphae and yeast-like cells, with a granulomatous tissual reaction. The isolated fungus was identified as Phoma cava. A review of the literature on fungal infection caused by different Phoma species, is presented. The patient healed after therapy with amphotericin B. followed by itraconazole. PMID- 9394537 TI - Presence of anti-Toxocara antibodies in children selected at Hospital Universitario, Campo Grande, MS, Brazil. PMID- 9394539 TI - Chronic chagasic cardiopathy: the product of a turbulent host-parasite relationship. AB - The pathogenesis of chronic chagasic cardiopathy is still a debated matter. In this review, the main theories raised about it since the first description of the disease in 1909 by Carlos Chagas, are considered. The scarcity of T.cruzi parasites into the myocardium and the apparent lack of correlation between their presence and the occurrence of myocardial inflammatory infiltrate, have originated many theories indicating that chronic Chagas' cardiopathy is an autoimmune disease. Recently however, papers using immunohistochemical technique or PCR have demonstrated a strong association between moderate or severe myocarditis and presence of T.cruzi Ags, indicating a direct participation of the parasite in the genesis of chronic chagasic myocarditis. Different patterns of cytokine production seem to have important role in the outcome of the disease. Participation of the microcirculatory alterations and fibrosis as well as the relationship with the parasite are also emphasized. Finally, the author suggests that the indeterminate form of the disease occurs when the host immunological response against the parasite is more efficient while the chronic cardiopathy occurs in patients with hyperergic and inefficient immune response. PMID- 9394538 TI - Possible role of Lutzomyia maranonensis and Lutzomyia robusta (Diptera: Psychodidae) as vectors of human bartonellosis in three provinces of region nor Oriental del Maranon, Peru. PMID- 9394540 TI - Effect of tannin-rich plant (Acacia nilotica) on some nutritional and bacteriological parameters in goats. AB - The effects of A. nilotica (tannin-rich plants) on the digestion coefficient of nutrients and its antibacterial properties were "in-vivo" investigated in this study. Twenty five male baladi goats aged 3-4 years and weighing 19-21 kg were randomly classified into five groups (5 per each). Each animal group was fed on one of the experimental diets containing different levels of A. nilotica leaves (0%, 5%, 10%, 15% & 20%). Chemical analysis of A. nilotica leaves revealed that its content of crude protein, crude fiber, ash were 12.1%, 30.5%, 13.2%, respectively, while the total soluble tannins were 34%. The digestion coefficient of dry matter (DM), crude protein, crude fiber and nitrogen free-extract (NFE) in the control group were better than those of the experimental diets. However, digestion coefficient of most nutrients were significantly decreased (P < 0.01) with increasing levels of A. nilotica in the goat's diets. Absorbed and retained nitrogen (g/day) were decreased with increasing levels of A. nilotica. Concerning nitrogen balance, all animals of the experimental groups showed positive nitrogen balance. Colony forming units (CFU) were drastically reduced in both faecal und ruminal juice samples. This reduction was directly proportional to levels of A. nilotica in the ingested feed. The CFU were reduced in the faecal samples from 5.9 x 10(8)/g (control group) to 2.8 x 10(4) (fifth group) (that received 20% Acacia nilotica). On the other hand, the CFU/ml of ruminal juice samples were reduced from 5.8 x 10(6) (control group) to 1.8 x 10(3) (fifth group). On the other hand, Cl. perfringens count was reduced from 1.0 x 10(4) to 6.3 x 10/g in the faecal samples while their count was reduced from 6.2 x 10(2) to 4.4 x 10/ml in case of ruminal samples. The reduction of total bacterial and Cl. perfringens counts were directly proportional to the levels of A. nilotica in the diets of goats. PMID- 9394541 TI - Differentiation of proteinase (minor toxin lambda) in Clostridium perfringens strains from sheep and goats in Iran. AB - 17 Clostridium perfringens strains isolated post mortem from sheep and goats in Iran were examined by biochemical tests and by enzyme immuno assay (EIA). 7 of the C. perfringens strains belonged to type B, 8 strains were type D, one strain was type A, one strain was untypable by EIA. The isolated strains were examined for presence of Minor Toxin Lambda (proteinase) to identify the Iran subtype of C. perfringens type B. The results are compared to characteristics of Clostridium perfringens reference strains. The differentiation technique for proteinase (Minor Toxin Lambda) is discussed. PMID- 9394542 TI - [Atopic myelitis: a novel clinical entity]. PMID- 9394543 TI - [Studies on the taste perceptive threshold for 4 basic taste qualities at various sites of lingual surface]. AB - Using filter-paper-disk method for 20 approximately 23 year-old 60 healthy females, taste perceptive thresholds for 4 basic taste qualities (sweetness, saltiness, sourness and bitterness) were measured detailedly at various sites of the lingual surface (lingual apex, lingual center, lingual margin, lingual radix). Sucrose, sodium chloride, sodium tartarate and quinine hydrochloride as taste-producing substances were used for the sweetness, saltiness, sourness and bitterness, respectively. The results revealed the apical site to be most sensitive to all of the sweetness, saltiness and sourness (and especially to the saltiness) and to be followed by marginal and radical sites in sensitiveness. The radical site was revealed to be most sensitive only to the bitterness, and the central site was remarkably hyposensitive to all of these taste qualities. These experimental results differ, in many respects, from those results of study which have been published by Kiesow in 1894 and are already established today (lingual sensitivity to taste qualities is found separately by site: sweetness, sourness and bitterness and highly perceptible at apical, marginal and radical sites, respectively, and saltiness is not very differently perceptible by site), indicating that further detailed reexamination will be needed to know how taste perceptive thresholds for 4 basic taste qualities are distributed in the lingual surface. PMID- 9394544 TI - [Regional pulmonary function by 133Xe gas and 99mTc-MAA in 11 cases with lymphangiomyomatosis (LAM)]. AB - The evaluation of the regional distribution of ventilation and perfusion was performed by 133Xe gas and 99mTc-MAA in 11 patients with LAM. We divided the lung images into six lung regions, the upper, middle, and lower lung fields of the left and right lungs, and classified the ventilation distribution pattern as one of three types according to the washout time. Prolongation of mean transit time (MTT) predominantly in the lower lung field was classified as type a, predominantly in the middle and upper lung fields as type b, and diffuse prolongation of MTT throughout the lung as type c. The classification included 16 cases of type b, four of type a, and two of type c. The 133Xe washout was predominantly delayed in the middle and upper lung fields in 73% of LAM cases. Pulmonary perfusion was reduced in the middle lung field and relatively increased in the lower lung field in comparison with healthy controls. A follow-up study of 133Xe gas lung scan was performed in three cases of type b. All the cases deteriorated and presented obstructive and restrictive disturbances without changes in the distribution pattern. These findings suggested that the washout type did not change with the progress of the stage of the disease. PMID- 9394545 TI - [Scatter correction with an off-peak triple energy window method in thallium-201 imaging]. AB - For scatter correction using the triple energy window (TEW) acquisition in 201Tl imaging, we propose an off-peak TEW (OFPTEW) method. This OFPTEW method employs a wide main energy window of 34 keV centered at 73 keV and two 5.1 keV sub-energy windows and uses the scatter correction factor of 0.55. To assess scatter correction using the OFPTEW method in 201Tl imaging, phantom studies for planar and SPECT imaging were performed and the data with the OFPTEW method were compared with those by the conventional TEW method using the trapezoidal formula with the 20% main energy window centered at 70 keV and two 4.9 keV sub-energy windows. The planar images corrected by both methods were visually similar. The OFPTEW method, however, estimated the true primary counts and the contrast value in the cold lesion accurately, while the conventional TEW method underestimated the primary counts by 30% and gave wrong contrast values. For the myocardial SPECT imaging, the short-axis images by both methods were very similar, but the images by the OFPTEW method had 1.46 times more counts than those corrected by the conventional TEW method. In conclusion, the OFPTEW method can correct scatter in 201Tl imaging accurately and increase the primary counts effectively compared with the conventional TEW method. PMID- 9394546 TI - [123I-MIBG lung uptake in patients with diabetes mellitus]. AB - The purpose of this study is to clarify the relationship between 123I-MIBG lung uptake and silent myocardial ischemia (SMI), cardiac autonomic neuropathy (AN) or clinical characteristics. For the quantitative analysis, lung to upper mediastinum uptake ratio (L/M) and heart to upper mediastinum uptake ratio (H/M) were obtained from chest planar image. In addition, both lung washout ratio (%WR L) and heart washout ratio (%WR-H) were calculated from early and delayed images. Each indices were compared in both diabetic and control groups. Mean values of H/M in diabetes with complication were significantly lower than those of control group. Particularly, AN(+)SMI(+) group showed lowest value. Similarly, mean values of %WR-H in diabetes with complication were significantly higher than those of control group and AN(+)SMI(+) group showed highest value. Although mean value of L/M in each diabetic group was significantly higher than that of control group, there was no statistical significance among each diabetes except AN(+)SMI( ) group on early image. Mean value of %WR-L in AN(+) or SMI(+) group was also significantly higher than that of control group, but there was no statistical significance among each diabetic group. The current study suggested that high pulmonary 123I-MIBG uptake in diabetes was independent of the complication of SMI or AN. Pulmonary endothelial dysfunction related with severity of diabetes mellitus was considered to be the most important factor. PMID- 9394547 TI - [Clinical significance of 123I-MIBG myocardial scintigraphy in patients with hypertrophic cardiomyopathy]. AB - We studied the abnormality of myocardial sympathetic nervous system in patients with hypertrophic cardiomyopathy using 123I-metaiodobenzylguanidine (MIBG) myocardial scintigraphy in comparison with the parameters of other clinical examinations. In 50 patients with HCM, the heart to mediastinum 123I-MIBG uptake ratio (H/M) was significantly low and washout rate (WR) of 123I-MIBG was significantly high respectively compared with normal subjects (n = 8). H/M was negatively correlated with serum norepinephrine level, wall thickness of left ventricle, left ventricular mass index, left ventricular end diastolic pressure respectively, and WR was positively correlated with those parameters respectively. On the other hand, LF/HF calculated by spectral analysis in holter electrocardiogram was positively correlated with H/M, and negatively correlated with WR. In HCM, H/M in patients with subjective symptoms was significantly lower than that without subjective symptoms, and WR in patients with paroxysmal atrial fibrillation was significantly higher than that without paroxysmal atrial fibrillation. This study revealed that H/M and WR reflected the severity and the difference of disease type in HCM. In conclusion, 123I-MIBG contributes to evaluating more details in diagnosis and pathophysiology of HCM. PMID- 9394548 TI - [Oligomelanotic malignant melanoma in the nasal cavity diagnosed by IMP scintigraphy]. AB - MRI, 67Ga scintigraphy, and 123I-IMP-SPECT were performed in the patient with oligomelanotic malignant melanoma in the nasal cavity which is not confirmed pathologically at first. MRI failed to diagnose the tumor as malignant melanoma. It was difficult to differentiate malignant melanoma from malignant lymphoma in 67Ga scintigraphy. The remarkable accumulation of 123I-IMP was consistent with the tumor localization in the nasal cavity. This tumor uptake was thought to be oligomelanotic or amelanotic melanoma since the accumulation was more distinctive at the delayed image and since the tumor was not visibly melanotic. Finally the tumor was confirmed to be oligomelanotic melanoma by immunohistochemical examination; which was in accordance with IMP-SPECT findings. Oligomelanotic or amelanotic melanoma occurs in nasal cavity with high frequency. We report here the oligomelanotic melanoma case where IMP-SPECT was rather useful to make a pathological diagnosis than other imaging modalities. PMID- 9394549 TI - [A case of an adult neuroblastoma with bone-marrow metastases: 131I-MIBG scintigraphy in comparison with MRI]. AB - To detect a primary neuroblastoma lesion and its metastases, 131I-MIBG scintigraphy was performed for a 24-year-old woman who had a high level of serum catecholamine. 131I-MIBG scintigrams showed high radioactivity in the left upper quadrant, pelvic bone, and vertebral bodies. A biopsy of the pelvic bone revealed metastasis from the neuroblastoma. After four chemotherapy courses, the accumulation of 131I-MIBG decreased after each course; however, scintigraphy performed after the last chemotherapy course showed focal mild uptake in the right sacroiliac. The presence of residual tumor in the sacroiliac was confirmed histologically. On the other hand, T1-weighted and T2-weighted MR images performed before the treatment showed low signal intensity and high signal intensity in the pelvic bone, respectively. After the fourth chemotherapy course, T2-weighted MR images showed low signal intensity in the pelvic bone; however, it was difficult to determinate whether it should improve. To assess the effect of treatment of neuroblastoma, 131I-MIBG scintigraphy was considered more useful than MRI. PMID- 9394551 TI - [Experimental study on the location of energy windows for scatter correction by the TEW method in 201Tl imaging]. AB - To investigate validity of scatter correction by the TEW method in 201Tl imaging, we performed an experimental study using the gamma camera with the capability to perform the TEW method and a plate source with a defect. Images were acquired with the triple energy window which is recommended by the gamma camera manufacturer. The result of the energy spectrum showed that backscattered photons were included within the lower sub-energy window and main energy window, and the spectral shapes in the upper half region of the photopeak (70 keV) were not changed greatly by the source shape and the thickness of scattering materials. The scatter fraction calculated using energy spectra and, visual observation and the contrast values measured at the defect using planar images also showed that substantial primary photons were included in the upper sub-energy window. In TEW method (for scatter correction), two sub-energy windows are expected to be defined on the part of energy region in which total counts mainly consist of scattered photons. Therefore, it is necessary to investigate the use of the upper sub-energy window on scatter correction by the TEW method in 201Tl imaging. PMID- 9394550 TI - [Evaluation of 123I-MIBG clearance from the myocardium; comparison of two methods -SPECT & planar methods]. AB - In 123I-metaiodobenzylguanidine (MIBG) scintigram, MIBG clearance from the heart is used to evaluate the severity of various heart diseases. There are two methods for calculating MIBG clearance. One involves planar images (planar method) and the other uses a bull's eye map (SPECT method). In 158 patients and 10 normal subjects, we compared these two methods. Fifteen minutes and 4 hours after intravenous injection of 111 MBq MIBG, planar images and SPECT images were obtained. Then clearance from the heart was calculated by each method. Abnormal increase was defined as present if clearance was more than the mean + standard deviation of 10 normal subjects. Then, we examined the sensitivity with which each method could detect clearance abnormality in 158 patients. Thirty-two patients showed abnormality only on SPECT images, while planar images alone showed abnormalities in only 5 patients. The reason the SPECT method was more sensitive than the planar method may be as follows; in the case of decreased MIBG clearance from the lung, for example, in congestive heart failure, clearance by planar method is apparently decreased. Thus, the SPECT method can detect clearance abnormality more sensitively than the planar method, and if we evaluate MIBG clearance from the heart by the planar method, we must take into account MIBG clearance from the lung. PMID- 9394552 TI - [Typical normal cases and normal cases with abnormal image pattern in every myocardial SPECT radiopharmaceutical]. AB - Working group of cardiac nuclear medicine was made as a Japanese part of society of international cardiac nuclear medicine under the cooperation between Japanese society of nuclear medicine and Japanese society of cardiology. We investigated typical normal cases and normal cases with abnormal image pattern in every myocardial SPECT radiopharmaceutical as one of the research activity of working group. From 11 faculties, 16 T1 cases, 14 BMIPP cases, 8 MIBG cases, 8 MIBI cases and 14 tetrofosmin cases were submitted as typical normal cases, and 12 T1 cases, 5 BMIPP cases, 12 MIBG cases, 10 MIBI cases and 5 tetrofosmin cases were submitted as normal cases with abnormal image pattern. We summarized the condition of SPECT data acquisition of each faculties. And we added the discussion from literature about how to discriminate normal cases with abnormal image pattern from abnormal cases. In MIBG, patterns of typical normal cases and normal cases with abnormal image pattern were slightly different from other 4 pharmaceuticals. In other 4 pharmaceuticals, diaphragmatic attenuation, breast attenuation, apical thinning and others were presented as normal cases with abnormal image pattern. PMID- 9394554 TI - [Evaluation of effective treatment drugs against Acanthamoeba cyst]. AB - Cysts of 2 isolates of Acanthamoeba from the cornea of 2 patients with confirmed Acanthamoeba keratitis were tested in vitro for sensitivity to antimycotic agents such as fluconazole, miconazole, amphotericin-B, pimaricin, antiprotozoal agents such as pentamidine isetionate and antiseptics which could be use in the ophthamological region. Pimaricin was the most successful cysticidal agent against the two strains. Sensitivity to pentamidine isetionate showed variation. Fluconazole, miconazole and amphotericin-B were resistant against cysts with concentration of eye drops that have been used in the treatment of Acanthamoeba keratitis. It was supposed that 5% pimaricin eye drops could be use in the treatment of Acanthamoeba keratitis in addition to keratomycosis. Pentamidine isetionate which belong to the diamidine family, is not yet clear as to the side effects to corneal epithelium cell, but we believe that this drug could be expected as a new therapeutic agent for Acanthamoeba keratitis. PMID- 9394553 TI - [Trend of bacterial meningitis in children over a 14 year period (1981 through 1994) in Japan--an analysis based on studies in 27 institutions]. AB - We observed 266 children with purulent meningitis in 27 institutions in Japan during the 14 years from 1981 on dividing these years into 3 periods, 1981-1985, 1986-1990 and 1991-1994, and studied the trend of causative organisms identified in 254 among the 266 patients. Their ages were less than 3 months after birth in 50 children and 3 months or older in 216: there were 141 boys and 125 girls. The causative organisms were H. influenzae in 134 patients and S. pneumoniae in 50, most of them being aged 3 months or older. Next to the above bacteria ranked S. agalactiae in 29 and E. coli in 12, many of the patients were aged less than 3 months. Staphylococcus spp. was found in 7 patients and about 70% of them were aged 3 months or older. L. monocytogenes was found in 4 patients and N. meningitidis in 3 and they were aged 3 months or older in both patient groups. S. pyogenes, Enterococcus spp., Peptostreptococcus spp., P. Mirabilis and Enterobacter spp. were detected each in 1 patient. The causative organism was unknown in 21 patients and there was no double infection. H. influenzae were detected in 18 patients in 1981-1985 period (36.7%), in 56 in 1986-1990 (54.9%) and in 60 in 1991-1994 (63.8%) showing an increasing tendency, but S. pneumoniae exhibited neither an increasing nor decreasing tendency. There was a decreasing tendency with S. agalactiae and E. coli, but the details were not clear because there were few patients aged less than 3 months. Although the period of coexistence of 4 main bacterial species was not made clear in this study. Listeria is considered to develop mainly in the early childhood, and we believe that the conventional way of using a cephem preparation and ampicillin combined for patients under 6 years need not be altered. However, panipenem (phonetic) is likely to be effective for insensible S. pneumoniae for the time being. PMID- 9394555 TI - [Clinical features of Japanese children infected with Cryptosporidium parvum during a massive outbreak caused by contaminated water supply]. AB - A massive outbreak of cryptosporidiosis occurred at a local town of Saitama Prefecture, in 1996. During this outbreak, we investigated the clinical features of children seen at Saitama Medical School. Cryptosporidium parvum (C. parvum) was detected from 10 out of 28 (36%) children with diarrhea during June and August, 1996. The average ages of the children who were positive and negative for C. parvum were 6.5 and 5 year old, respectively. Among the children infected with C. parvum, colic pain was observed in 3 children and 4 children had vomiting. However, none of the children showed fever over 38.0 degrees C nor bloody stools. Family members of children infected with C. parvum also had diarrhea and/or vomiting (5/6). C. parvum was repeatedly detected from 2 out of 3 children. All infected children had an improvement of abdominal symptoms in 4 to 10 days. C. parvum should be included as a pathogen which causes enterocolitis in Japanese children. PMID- 9394556 TI - [Distribution of serogroups of Vibrio cholerae non-O1 non-O139 with specific reference to their ability to produce cholera toxin, and addition of novel serogroups]. AB - A total of 1898 strains of Vibrio cholerae non-O1 non-O139, which had been collected worldwide for the past 3 year period of 1994-1996, were serogrouped. The strains were also examined for presence of cholera toxin (CT) gene (ctx) and NAG-ST gene, and strains which carried to ctx were further analyzed for their ability to produce CT. In addition, attempts were made to establish novel serogroups for those serologically untypable strains. Of those examined, 1,774 strains of V. cholerae non-O1 non-O139 was classified into 128 known serogroups while 50 strains were found to belong to R type, and the rest of the 74 strains could not be serotyped. Distribution of the serogroups did not seem to correspond to either the strains geographic distribution or sources of isolation. Of those serologically untypable strains, 38 novel serogroups (O156-O193) were established and added to our reference of V. cholerae antigenic schema. It was also found that antisera raised against many V. cholerae strains included R antibodies. This indicates that any V. cholerae antisera for diagnostic purpose should be absorbed with the reference R strains, CA385, before use. There were luminescence producing strains among those sucrose and VP reaction negative strains. Subsequent DNA/DNA homology analysis revealed that they were identified as V. cholerae. This points to a possibility that strains tentatively identified as Vibrio mimicus by conventional biochemical tests may have included luminescent strains of V. cholerae. It is thus highly recommended that strains in question should be tested for the luminescence production in order to differentiate V. cholerae from V. mimicus. Of those 1989 strains examined, 37 strains (ca. 2%) were found to produce CT. Interestingly, CT producing strains were prevalent in serogroup O141; 10 strains out of 16 strains (63%) were positive for CT. The evidence calls for a caution to possible occurrence of cholera-like diarrhea caused by V. cholerae O141 in the future. PMID- 9394557 TI - [Diagnosis of neonatal enterovirus meningitis by reverse transcription-polymerase chain reaction]. AB - We evaluated the potential clinical utility of a reverse transcription-polymerase chain reaction (RT-PCR) for neonatal enterovirus meningitis, comparing the results with viral culture and white blood cell (WBC) counts in the cerebrospinal fluid (CSF). Of the 41 cases of enteroviral infection, 31 cases (76%) were finally diagnosed as meningitis by either viral culture, CSF WBC count or RT-PCR. Of those with culture positive and negative, the RT-PCR positive rates were 10 (100%) and 6/13 (46%), respectively. Of those with and without WBC increase in the CSF, the RT-PCR positive rates were 18/20 (90%) and 5/14 (36%), respectively. There was one RT-PCR positive case (1/6, 17%) among those with culture negative and no WBC increase in the CSF. RT-PCR performed on CSF is a sensitive and specific method for the diagnosis of neonatal enterovirus meningitis. PMID- 9394558 TI - [Effect of the prior influenza vaccination on serum antibody titer induction by subsequent inactivated influenza vaccine in the elderly]. AB - In order to investigate the effects of prior influenza vaccination on subaequent annual influenza vaccination in the geriatric population, we analyzed serum hemagglutinine inhibition antibody tirers (HI titer) before and after vaccination with inactivated influenza vaccine in elderly inpatients. A total of 163 inpatients of 60 years or older were enrolled with informed consent. They were classified by vaccination status in the previous year, 53 patients had inactivated vaccine (inactivated). 52 patients had genetically assorted cold adapted influenza live attenuated vaccine (cold-adapted), and 53 had no influenza vaccine history during the past year. The HI titer was higher in the inactivated group than in the cold-adapted and non-vaccinated groups, suggesting residual immunological effects of inactivated influenza vaccine from the previous year vaccination. The HI titer after the inactivated vaccine in 1993 was higher in both the inactivated and cold-adapted groups than in the non-vaccinated group. The number of patients with HI titers of 2(7) or higher, which is the putative protective HI titer level for influenza infection, was significantly higher in both the inactivated and cold-adapted groups than the non-vaccinated group. These results suggest that continuous annual influenza vaccination does not impair the effects of vaccination, and may actively promote elevated HI titers. PMID- 9394559 TI - eaeA genes in Escherichia coli derived from Japanese patients with sporadic diarrhea. AB - To investigate the prevalence of attaching and effacing Escherichia coli, we examined 364 strains isolated from the feces of 9,684 patients with diarrhea at the Anjo Kosei Hospital in Japan for the presence of eaeA. Twenty-nine (8%) of the strains were eaeA positive. Of enteropathogenic E. coli (EPEC), 11 of the 87 (13%) strains were for the positive eaeA gene. The serotypes and the numbers of eaeA-positive strains among the strains tested were as follows: O26:H-(2/3), O55:H7 (4/4), O55:H-(2/ 2) and O128:H2(3/3). Two enterohemorrhagic E. coli (EHEC) strains (Verotoxin positive O157:H7) were also eaeA positive. Among 260 non-EPEC strains that were not categorized as diarrheagenic E. coli, 16 (6%) were eaeA positive. Those serotypes were as follows: O15:H2, O20:H6, O28:H28, O63:H6. O153:H7, O28:H6, O153:H19 and O157:H45. EPEC strains including O18:H7 and six other serotypes, enteroinvasive E. coli (EIEC), and enterotoxigenic E. coli (ETEC) were all eaeA negative. PMID- 9394560 TI - [Toxic shock-like syndrome after acupuncturation]. AB - A 80-year-olkd male was admitted to our hospital because of severe pain and swelling on his left lower leg on January 23, 1996. He had received an acupuncture to both legs because of intermittent claudication once a week from July, 1995 to January 18, 1996. On the next day of the last acupuncture, pain and swelling on his left leg appeared. On admission, his left leg showed diffuse swelling and redness with blisters. We diagnosed this patient as toxic shock-like syndrome (TSLS), based on the rapid exacerbation of the skin changes, necrotizing superficial fasciitis, multiple organ failure with shock, and the detection of group A streptococcus from culture samples obtained from both skin blister and necrotic fascia. He recovered from the disease by amputation of the involved leg and antibiotic therapy. Acupuncture could have been the cause of streptococcal infection. PMID- 9394561 TI - [Campylobacter jejuni enteritis in three patients with HIV infection]. AB - Gatrointestinal symptoms, which include diarrhea, are as common as respiratory symptoms in patients with HIV infection. Gastrointestinal symptoms may result from infections, neoplasma, HIV enteropathy or drug toxicity. Three HIV-infected patients admitted to our hospital complaining of diarrhea and fever. We confirmed their diagnosis as Campylobacter jejuni enteritis by bacteriological examination of their feces. All of them had eaten inadequately cooked meat in restaurants before the onset of their enteritis. Their symptoms immediately improved after the administration of antimicrobial agents. One strain of C. jejuni isolated in our cases, however, was resistant to ofloxacin. This case report suggests that we must counsel HIV-infected patients to avoid inadequately cooked food and observe resistant patterns of C. jejuni to antimicrobial agents in Japan in the future. PMID- 9394562 TI - [A case of cat scratch disease identified by an elevated Bartonella henselae antibody level using enzyme immunoassay]. AB - A 68-year-old male was admitted to our hospital because of fever and a 2-week history of inguinal adenomegaly. Since he owned a cat, cat scratch disease was suspected. But it was necessary to distinguish cat scratch disease from lymphoma type adult T-cell leukemia because he showed a high level of antibody against HTLV-1. An excisional biopsy of the inguinal node was performed. Histopathologic examination revealed abscess-forming granulomatous lymphadenitis compatible with cat scratch disease. A Warthin-Starry silver stain showed pleomorphic bacilli in the lymph node. So we confirmed a serological response to Bartonella henselae, the causative agent of cat scratch disease, using enzyme immunoassay (EIA). The IgG antibody level to B. henselae was positive at 42 EIA Unit before treatment. After treatment with intravenous cefepime and oral tosufloxacin, his physical symptoms improved and the antibody level decreased to less than 12 EIA Unit. EIA was very useful for diagnosis of this case. Serology to B. henselae may replace traditional diagnostic criteria for cat scratch disease. PMID- 9394563 TI - [Acute respiratory failure associated with G-CSF-induced leukocyte recovery in three patients with preceding infection]. AB - Acute respiratory failure (ARF) occurred at the time of leukocyte recovery promoted by granulocyte colony-stimulating factor (G-CSF) in three patients with the preceding infection (S. aureus pneumonia, varicella zoster, and P. aeruginosa bacteremia, respectively) which had developed during leukopenia after cancer chemotherapy. G-CSF was used for 4 to 6 days, and the leukocyte counts at onset of ARF were 19,300/microliter, 11,300/microliter, and 4,100/microliter, respectively. All of the three patients received high-dose methylprednisolone and the artificial respiration was used in two. Consequently two patients responded well and survived, but one died of respiratory failure 2 weeks after occurrence of ARF. Autopsy of the dead case revealed mild interstitial pneumonia in the both lungs together with bacterial pneumonia in the right lobe. These cases indicate that G-CSF-induced leukocyte recovery can cause severe ARF in patients with precending infection. Therefore, G-CSF should be administered very carefully to granulocytopenic patients with infection. PMID- 9394564 TI - [Case report: lung abscess caused by Streptococcus agalactiae]. AB - Streptococcus agalactiae is a well-recognized cause of neonatal sepsis and meningitis. In adults, infections by S. agalactiae are rare. We report an adult case of lung abscess and pyogenic spondylitis caused by S. agalactiae. A 51-year old male was admitted to our hospital because of an abnormal shadow in the chest and lumbago on May 25, 1995. He was diagnosed as lung abscess from the chest roentgenogram and CT scan and the subcutaneous pus was aspirated. The pus culture was only positive for S. agalactiae. He was treated with IPM/CS 1 g/day and CLDM 1.2 g/day and the abscess was drained. MRI showed his lumbago was caused by pyogenic spondylitis. The underlying disease of this case was diabetes mellitus. He recovered from the infections with in about 10 weeks of antibiotic treatment. PMID- 9394565 TI - DNA diagnosis of Plasmodium falciparum malaria by single-tube PCR. PMID- 9394566 TI - [An evaluation of computed radiography in contrast-enhanced imaging of the gastric area]. AB - The usefulness of computed radiography (CR) in contrast-enhanced imaging of the stomach was evaluated. First, the spatial resolution and density resolution of CR under the conditions of contrast-enhanced imaging of the digestive tract were examined. Spatial resolution was lower, but density resolution was similar, in comparison with those of conventional film screen (FS). Both were better in CR at a low dose. Evaluation of processing conditions for CR using a gastric phantom indicated that the use of N for gamma type (GT) and T for response type (RT) in the left image and the use of A for GT and F for RT in the right image is recommended. Evaluation of the relationship between the stomach segment and response enhancement (RE) showed that RE should not be very strong in cases where segments to be visualized are limited to the lower part of the stomach. With an optimal RE, the image quality of the gastric segment was better in CR than in FS. When dynamic-range control processing was performed in an area overlapped by colon gas, obscure parts became visually recognizable. CR is considered to be useful also for contrast-enhanced imaging of the digestive tract. PMID- 9394567 TI - [Evaluation of therapeutic effect using enhanced MRI in lung cancer: evaluation of methods in terms of necrosis]. AB - To evaluate therapeutic effect in terms of necrosis or cavity, enhanced MRI was performed in 40 lung cancer patients treated by conservative therapy. We provided the reduction ratio of the viable tumor as calculated by a volume method and a cross-sectional method. In the volume method, the volume of necrosis was subtracted from the volume of the tumor, and in the cross-sectional method, the product of the longest diameter and widest perpendicular diameter of necrosis was subtracted from the product of the longest diameter and widest perpendicular diameter of the tumor. We then examined whether we could substitute the cross sectional method for the volume method. The reduction ratios of viable tumor calculated by the two methods were in good correlation. The limits of agreement of each method and their repeatability coefficients were considered small enough for clinical use. Therefore, we concluded that the cross-sectional method could be used in place of the volume method for clinical purposes. In evaluating therapeutic effect in terms of necrosis when using contrast-enhanced MR imaging, the reduction ratio of the viable tumor determined by the cross-sectional method can be substituted for that determined by the volume method. PMID- 9394568 TI - [Usefulness of photon counting X-ray radiography for diagnostic imaging of the thorax: experimental and clinical studies]. AB - To evaluate the usefulness of photon counting X-ray radiography (quantum radiography, QR) for diagnostic imaging of the thorax, comparative studies between QR and the conventional screen-film system (SF) were performed. Exposure dose, spatial resolution and density resolution on QR were evaluated in the experimental study. The ability of QR to describe normal structures and several kinds of abnormal shadows was evaluated in the clinical study. The relative exposure dose of QR was lower than that of SF, while the spatial resolution of QR was slightly inferior to that of SF. However, the density resolution of QR was superior to that of SF. Clinically, QR was superior to SF in the description of normal structures and several kinds of abnormal shadows. In conclusion, it was considered that QR is useful for diagnostic imaging of the thorax. PMID- 9394569 TI - [Radiation therapy for adrenal metastases]. AB - PURPOSE: To evaluate the role of radiation therapy for adrenal metastases. MATERIALS AND METHODS: Fourteen patients, 13 with primary lung carcinoma and one with primary unknown carcinoma, received radiation therapy for adrenal metastases from 1984 to 1995 at the Hyogo Medical Center for Adults. Total dose ranged from 16 Gy to 60 Gy, and fractional dose from 1.6 Gy/ Fr to 3 Gy/Fr. RESULTS: Partial response of the local tumor was recognized in 2 of 7 patients by CT imaging. Pain relief was obtained in 7 of 8 patients. Median survival was 3 months, and 6-month survival was 28.6% in all patients. Among patients in the symptomatic group, who had complaints of pain due to adrenal mass, survival was even worse (12.5% at 6 months). There were no severe complications, but 4 patients (29%) had gastrointestinal symptoms. CONCLUSION: Radiation therapy is useful for the purpose of pain relief in adrenal metastases. PMID- 9394570 TI - [Radionuclear estimation with 99mTc-N-pyridoxyl-5-methyl-tryptophan (PMT) scintigraphy of recurrent hepatocellular carcinoma after non-surgical treatment]. AB - 99mTc-N-pyridoxyl-5-methyl-tryptophan (PMT) scintigraphy, which exhibits highly specific for the diagnosis of primary hepatocellular carcinoma (HCC), was performed for recurrent HCC following non-surgical treatment. Eighty-four patients (total 351 examinations) were selected for the study. The ability of PMT scintigraphy to diagnose recurrent HCC after treatment was compared with that of computed tomography, ultrasonography and angiography. The results showed that 1) sixty-three (75%) of 84 cases demonstrated positive findings during our follow-up period, 2) most of the tumors initially showing positive findings remained PMT positive when they recurred and 3) PMT studies shortly after treatment demonstrated wider tracer uptake rather than the actual tumor area. In conclusion. PMT scintigraphy for patients with HCC after non-surgical treatment exhibited high accuracy in identifying viable HCC only for tumors indicating positive findings. PMID- 9394571 TI - [CT-guided lung needle biopsy using a new supporting implement]. AB - We contrived a new supporting implement to increase the hit rate of CT-guided needle biopsy (CTNB) for localized pulmonary lesions. Using this implement, twenty-two CTNB examinations for localized pulmonary lesions were performed. In 21 out of the 22 examinations (95%), the lesions were hit, and specimens appropriate for cytological or histological diagnosis were obtained. The course of needle insertion, which was difficult to define in the past, has become to be easier and more precise with this method. Using this new implement, CTNB is now applicable to smaller and deeper lesions. PMID- 9394572 TI - [Radioresistant CD4+ T cells in normal, unprimed mice: with verification of the Bergonie-Tribondeau law]. AB - This is the first report on radioresistant CD4+ T cells found in normal, unprimed mice. After sublethal whole body irradiation, regular CD4+ as well as primitive NK1.1 + CD4+ T cells were enriched in the spleen. Since it has been well established that virgin T and B cells are highly radiosensitive, these cells were once assumed to be a unique lymphocyte population for which radiosensitivity does not follow the general law of radiation sensitivity for mammalian cells (Bergonie Tribondeau law). These cells exhibited higher proliferative response to accessory cells than the non-irradiated control cells in the syngeneic mixed leukocyte reaction (SMLR). This indicated that virgin CD4+ T cells sensitized to, and readily respond to self-MHC class II molecules are radioresistant, and that their radioresistance, as activated cells, is consistent with the Bergonie-Tribondeau law. PMID- 9394573 TI - [Cardiac surgery in patients with chronic renal failure on maintenance dialysis]. AB - From July 1988 through August 1996, 54 patients with chronic renal failure (CRF) on maintenance dialysis (50 hemodialysis = HD, and 4 continuous ambulatory peritoneal dialysis) have undergone some sort of surgical procedure requiring the use of extra corporeal circulation (ECC); 42 patients underwent isolated coronary artery bypass grafting (CABG), 8 valve replacement, 3 combined procedures and 1 correction of a congenital heart defect. The protocol called for maintenance dialysis on the day before surgery, large volume hemofiltration (HF) during the ECC period, postoperative K+ management with dextrose-insulin if required, and resumption of whatever preoperative maintenance dialysis 24 hours after the operative procedure. The mean diafiltrate volume of HF was 7963 +/- 2688 ml which was replaced with 6342 +/- 2748 ml. No patient required emergency HD before the resumption of the maintenance dialysis, although in 40% of the early patients HD was added on the second postoperative day. However as experience was gained, in the latter 60% of patients resumption of maintenance dialysis (HD 3 times a week) was thought to be sufficient. The incidence of calcification in patients with CRF is higher not only of involved coronary artery segments (4.5 +/- 2.3 segments; AHA coronary classification) than its counterpart without CRF, but also of the ascending aorta which mandated modifications of the technique in 6 patients (operation under ventricular fibrillation, cannulation access other than ascending aorta). The use of arterial in situ conduits for CABG was also thought to be advantageous, and the left internal thoracic artery combined to the gastro epiploic artery was used in 11 patients (26.2%). Four patients died) (7.4%): 2 from arrhythmia, one from intestinal necrosis and one from multiple cerebral infarction. Thus we conclude that the outlined protocol is quite effective in controlling fluid and electrolyte balance in patients on maintenance dialysis allowing to undertake surgical procedures requiring the use of extra corporeal circulation relatively safely. PMID- 9394574 TI - [Glutamate neurotoxicity during spinal cord ischemia--development of a delayed onset paraplegia model]. AB - The incidence and severity of spinal cord dysfunction are related to both the depth and duration of the resulting ischemic state. Evidence is accumulating that glutamate, a major neurotransmitter, has potent neurotoxic activity during ischemia. In our laboratory, it has been confirmed that exogenous glutamate has detrimental effects on spinal cord neurons during brief ischemia in vivo. We hypothesized that glutamate neurotoxicity is associated with delayed-neuronal dysfunction. Delayed-onset paraplegia is defined as a neurologic deficit which develops after initial recovery. Infrarenal aortic segments from 12 New Zealand white rabbits, were isolated for 5 minutes and perfused at a rate of 2 ml/min. Group I (n = 6) received normothermic saline (39 degrees C). Group II (n = 6) received normothermic L-glutamate (20 mM). Neurologic function was assessed at 6, 24, and 48 hours after surgery according to the modified Tarlov scale. After 48 hours, the rabbits were euthanized and their spinal cords were harvested for histologic examination. The neurologic function of all group I was fully intact, whereas three rabbits in group II showed acute paraplegia and the other three showed delayed-onset paraplegia. Histologic examination of spinal cords from rabbits in group I revealed no evidence of cord injury, whereas spinal cords from those in group II had evidence of moderate spinal cord injury with central gray matter and adjacent white matter necrosis and axonal swelling. These results indicate that dose-dependent glutamate neurotoxicity is associated with delayed neuronal dysfunction following ischemia in vivo. The severity of the ischemic event, i.e., extracellular glutamate overload, is suspected to be the etiology of delayed-onset paraplegia which, in turn, is thought to be the result of borderline ischemia. This model may allow a pharmacologic approach to the prevention of ischemic spinal cord injury. PMID- 9394575 TI - [Radial artery for coronary artery bypass graft]. AB - Early postoperative results were studied in 50 cases of coronary artery bypass graft (CABG) using a radial artery (RA). The patients ranged in age from 37 to 81 years, with the mean age of 61 years. Of them, 49 were male. An average of RA was 17.6 cm at completion of detachment and 15.6 cm when the graft was cut for use. The internal diameter before anastomosis an average of 3.7 mm on the proximal side and an average of 2.8 mm on the distal side. RA was anastomosed with ascending aorta in 47 cases, with the left internal thoracic artery in 2 cases and with the right internal thoracic artery in one case on the proximal side. RA was anastomosed with the left anterior descending branch area in 6 cases, with the left circumflex branch area in 40 cases and with the right coronary artery area in 4 cases on the distal side. There was no case of operative death, but one patient died while in hospital. The cumulative patency rate of the RA grafts was 95% (n = 40). Early postoperative results of the RA graft were satisfactory, therefore the RA graft were satisfactory, therefore the RA graft was an excellent alternative conduit for myocardial revascularization. PMID- 9394576 TI - [Brain damage after surgery for thoracic aortic aneurysm]. AB - We analyzed cases with brain damage after surgery for thoracic aortic aneurysm in our institution and investigated the causes, risk-factors and preventive measures for this disastrous postoperative complication. Irreversible brain damage was a complication in 25 out of 184 operative cases (13.6%) over a 21-year period. The cause of brain damage was determined to be embolism by manipulation of the aorta in six cases, clamping of the left subclavian artery in four cases, technical problems of separate cerebral perfusion (SCP) in four cases, severe shock in three cases, embolism unrelated to operative maneuver in three cases, stenosis of a branch of the arch with aortic dissection in two cases, and air embolism, circulatory arrest with insufficient hypothermia and hypoperfusion of a temporary bypass to the left carotid artery in one case each. The neurological symptom improved in eight cases and was unchanged in 17 cases. Eighteen cases died in the hospital. In the univariate analysis, age (p = 0.048), a portion of the aneurysm (p = 0.035), preoperative brain complication (p = 0.003), emergency operation (p = 0.033) and clamping of the arch (p = 0.001) were found to be prominent risk factors for brain damage. In the multivariate analysis, clamping of the arch (p = 0.0310), SCP (p = 0.0327) and emergency operation (p = 0.0223) were prominent. To prevent postoperative brain damage, the arch should not be clamped, appropriate operative techniques to avoid bleeding and to shorten SCP time should be employed, and proper and prompt management of the emergency operation and caution in clamping the left subclavian artery are considered to be necessary. PMID- 9394577 TI - [Carinal resection for primary lung cancer--with special attention to a modified double-barrel method]. AB - Carinal resection for primary lung cancer was clinically evaluated. Carinal resection was performed in 18 patients, 17 males and one female, with a mean age of 64 years. Nine patients underwent carinal reconstruction and the other 9 sleeve or wedge pneumonectomy. The carinal reconstruction was of the montage type in one patient, the one-stoma type in 2, and the modified double-barrel method in 6. The modified double-barrel method is a technique that we developed by adding bronchial end-to-side anastomosis to the tracheobronchial end-to-end anastomotic site. A pedicled intercostal muscle flap was used for covering the anastomotic site. The postoperative respiratory complications after carinal reconstruction were anastomosis failure in 4 patients (pin-hole in 3) and respiratory failure in 2. However, no anastomosis stricture occurred, and recovery was satisfactory. There were no respiratory complications after pneumonectomy. One patient had renal failure before surgery and died of multiple organ failure 23 days after a montage type carinal reconstruction. The other 17 patients did well and could be discharged from the hospital and the overall mortality rate was 5.6%. No anastomosis stricture occurred in the modified double-barrel method. By carinal reconstruction covering of the anastomotic site is mandatory to prevent fatal postoperative complications. PMID- 9394578 TI - [Extra-anatomic bypass from the ascending aorta to the supraceliac abdominal aorta--surgical option applied to reoperation for aortic coarctation or interruption]. AB - The optimal approach for reoperation following repair of aortic coarctation (CoA) or interruption (IAA) remains controversial. Four patients underwent extra anatomic bypass for restenosis after repair of CoA or IAA. The age ranged from 4 to 12 years. The initial repairs for two CoA, one type A-IAA, and one type B-IAA consisted of two grafting, one subclavian arterial turning-down aortoplasty, and one subclavian flap aortoplasty. All of them underwent during infancy. Preoperative right arm systolic pressure ranged from 140 to 190 mmHg ar rest. Through a midline sternotomy and an upper laparotmy incision, an extra-anatomic bypass from the ascending aorta to the supraceliac abdominal aorta was employed using a 12 to 18 mm tube graft. All patients survived surgeries, and their hypertension markedly improved. Our experience confirms safety and effectiveness of this option in selected young patients with re-stenosis of following repair of CoA or IAA. PMID- 9394579 TI - [Composite valve graft replacement in patients with type A aortic dissection--a modified cabrol procedure]. AB - Composite valve graft replacement of the ascending aorta and aortic valve is indicated for a variety of conditions affecting the aortic root. However, a major drawback in this operation is bleeding from the proximal suture line and coronary anastomosis especially in patient with friable root tissue involved by aortic dissection. We describe here a modified technique to take advantage of the aortic button and cabrol techniques to reattach the coronary artery ostia. We have experienced seven patients with the aortic root replacements for type A dissection using the described technique over the past two years. In view of our favorable experience, we recommend this technique especially for patient with acute dissection involving nondilated aortic annulus, in addition to the patients with Marfan syndrome or annulo-aortic ectasia. PMID- 9394580 TI - [Lymph node dissection during a video-assisted lobectomy is inferior to that in a standard lobectomy]. AB - The indications for a video-assisted lobectomy are currently ill-defined. Clinicians recommend based on the extent of lymph node involvement. Fifty-nine patients with clinical stage I non-small cell lung cancer underwent lobectomies with systemic lymph node dissections through a standard thoracotomy (Group C), and 26 patients underwent lobectomies with lymph node dissections using the video assisted procedure (Group V). The number of dissected lymph nodes at all node levels were compared between the two groups. There was no significant difference between groups in the total number of dissected lymph nodes in patients with right lung cancer. The number of dissected hilar and interlobar lymph nodes, however, was less in Group V than that in Group C (hilar: 1.2 +/- 0.4 vs. 2.8 +/- 0.6, interlobar: 1.1 +/- 0.4 vs. 2.1 +/- 0.4). The total number of dissected lymph nodes in patients with left lung cancer was significantly less in Group V than that in Group C (18.5 +/- 0.3 vs. 28.7 +/- 2.4). In addition, the number of dissected lymph nodes in pratracheal, pretracheal, tracheobronchial, subcarinal, hilar, and interlobar lymph nodes were significantly less in the group V than those in Group C. Although there was no significant difference in the actual survival rates between the groups in this preliminary study, a sufficiently small number of dissected lymph nodes in the video-assisted lobectomy may have resulted in inaccurate staging and poor prognosis in these patients. PMID- 9394581 TI - [Effects of aprotinin on blood loss reduction in children undergoing repair of tetralogy of Fallot]. AB - The effect of aprotinin on the blood loss reduction was studied in children undergoing repair of tetralogy of Fallot. We administered aprotinin to consecutive 21 patients during the repair of tetralogy of Fallot and examined the blood loss and operative time in comparison with that in a control group of 20 patients. 30,000 KIU/kg of aprotinin was infused as the initial cardiopulmonary bypass (CPB) dose and 10,000 KIU/kg/hr was continuously administered as the maintenance dose during CPB. There was no resternotomy case due to bleeding and no operative death in both groups. Blood loss after CPB during operation and total blood loss during operation were significantly lower in aprotinin group than in control group. There were no differences between two groups in the volume of chest tube drainage in the postoperative 24 hours and the duration of chest tube drainage. Time from cessation of CPB to skin closure and total operative time were significantly shorter in aprotinin group than in control group. In conclusion, aprotinin was effective on the reduction of blood loss and the shortening of operative time in children undergoing repair of tetralogy of Fallot. PMID- 9394582 TI - [A prospective study on the timing of discontinuation of aspirin before coronary artery bypass grafting]. AB - The effects of the timing of discontinuation of aspirin before coronary artery bypass grafting (CABG) on postoperative blood loss and blood requirements were examined in 22 patients undergoing elective CABG, who were randomly assigned into two groups. In Group I (11 patients), aspirin was discontinued two days before the operation and in Group II (11 patients), aspirin was continued up to the operation. The other 40 patients, who did not take aspirin for at least seven days before the operation, served as a control Group. There were no differences in preoperative data including the platelet count and the hemoglobin concentration, nor in operative variables such as operation time, cardiopulmonary bypass duration and aortic crossclamp time among the groups. Although postoperative blood loss (six hours' loss; Group I 218 ml, Group II 183 ml and control Group 172 ml) and red blood cells transfusion requirements were not different among the groups, platelet concentrates transfusion was more frequently required in Group II (54.5%) as compared with control Group (7.5%) and Group I (9.1%). The difference between Group II and the control Group reached statistical significance (p < 0.01), but there was no significant difference between Group I and control Group. This fact suggests that preoperative two days' discontinuation of aspirin works as effectively as seven days' discontinuation. PMID- 9394583 TI - [A clinical application of histidine buffered cardioplegia]. AB - Blood cardioplegia has been widely accepted due to better oxygen delivery, pH buffering and free radical scavenge. We have found that a crystalloid cardioplegia solution formulated to accelerate anaerobic glycolysis with high buffering capacity. To conserve blood cardioplegia, we formulated a crystalloid cardiopletia containing 100 mM histidine for buffering. This cardioplegia (HBS) was compared to cold blood cardioplegia in patients requiring open heart surgery. Eighty patients including HBS (n = 28), and CBC (n = 40) were involved in this study. Left ventricular end-systolic elastance (Emax; mmHg/cm3) was evaluated pre and postoperatively. Cardiac index and inotropic requirement were also monitored at 1, 3, and 12 hours after cardiopulmonary bypass. There was no death in either group. All hearts returned to previous rhythm in HBS group, whereas total 12 DC cardioversions were requested in 6 patients. Emax was significantly higher in HBS group (5.2 +/- 0.6 mmHg/cm3) than in CBC group (3.4 +/- 0.4 mmHg/cm3). Cardiac index was also significantly higher in HBS group postoperatively than in CBC group with lower inotropic requirements. We conclude that histidine containing crystalloid cardioplegia provides excellent recovery of cardiac performance with lower inotropic requirements in open heart surgery. The ease of use, and lack of blood are other important advantages of this crystalloid cardioplegia. PMID- 9394584 TI - [Assessment of right gastroepiploic artery grafts by thallium scintigraphy]. AB - From 1990 to 1996 we performed coronary artery bypass grafting using only arterial grafts. Both pre- and post-operative thallium-201 exercise myocardial scintigraphy were evaluated in 68 cases (161 grafts). The rate of improvement (%) was defined as follows: (number that showed improvement of perfusion of thallium on post-operative scintigraphy/number that showed decreased perfusion of thallium on pre-operative scintigraphy) x 100. Examination was made separately regarding cases of ischemia (102 grafts) and infarction (54 grafts). For ischemic cases, the rate of improvement using left internal thoracic artery (LITA), right internal thoracic artery (RITA) and right gastroepiploic artery (RGEA) was 80% (12/15), 70% (7/10) and 71% (5/7) respectively. For infarction cases, the rate of improvement using LITA, RITA and RGEA was 54% (7/13), 60% (6/10) and 53% (9/17) respectively. Among these three groups no significant differences were noted. As a result, RGEA is thought to have usefulness equivalent to LITA and RITA. PMID- 9394585 TI - [Thickness of the muscle layer of the gastroepiploic artery and the internal mammary artery--a presumable factor of flow instability in GEA during the perioperative period]. AB - Recently the right gastroepiploic artery (RGEA) is often used for coronary bypass grafting. Although patency rate of the RGEA is as high as that of the IMA, instability of blood flow through the RGEA during the perioperative period is reported. We assumed that the RGEA is more predisposed to spasm than the internal mammary artery (IMA). This study was carried out to verify the following two points. 1. The GEA has a smaller internal diameter and thicker muscle layer than the IMA. 2. The contractile force of the muscle layer of the GEA are stronger than those of the IMA under the same transmural pressure due to the greater thickness o the muscle layer of the GEA. METHODS: The RGEA was obtained at its full length from gastorectomy cases due to gastric cancer (n = 25). The distal section of the IMA was obtained from the left IMA during bypass grafting (n = 23). All specimens were stained by the Masson-trichrome method and examined microscopically. The thickness of the smooth muscle layer of the media and the internal radius were compared between the RGEA and the IMA. RESULTS: The thickness of the muscle layer was 274.0 +/- 13 microns in the RGEA, and 169.1 +/- 8 microns in the IMA (p < 0.01) that is the thickness in the GEA was 1.62 times greater in the IMA. Although a significant difference was not obtained, the internal radius of GEA (563.7 +/- 21.8 microns) was smaller than that of IMA (583.1 +/- 12.0 microns). Based on the internal diameter-elastic wall tension relationship and the Laplace law, internal diameter and elastic tension in both arteries were obtained at the same blood pressure. Mean elastic tension and internal diameter in the GEA were considered to be smaller than than those in the IMA. The values of the internal diameter of the arteries obtained from the theoretical view point were correlated well with those obtained by the histometoric methods. As the muscle layer of the arterial wall of the GEA is thicker than that o the IMA, and the internal diameter of the GEA tends to be smaller than that of the IMA, the stronger contraction o the muscle layer, when induced, would reduce the blood flow in much greater extent in the GEA than the IMA. CONCLUSION: These results support the assumption that the RGEA reacts strongly than the IMA to constructor agents and physical stimuli, thereby inducing a greater instability of blood flow. Therefore, RGEA grafts should be carefully handled during bypass grafting. PMID- 9394586 TI - [Four cases of adrenal tumor discovered through examination before surgery for lung cancer]. AB - Preoperative CT and Ultrasonography (US) showed adrenal tumors in four patients with lung cancer. Although metastasis of the cancer to the adrenal gland was suspected, a definitive diagnosis could not be made by CT and US alone. MRI is as ineffective as CT and US. Needle biopsy is useful if tumor cells are detected, but not unless they are discovered. Surgery, therefore, is necessary to establish the final diagnosis. (Adrenalectomy was performed on all cases, one of which had metastasis). No particular complications occurred after adrenalectomy. Adrenalectomy was considered unavoidable to determine stage and treatment policies in patients suspected of metastasis in imaging diagnosis. PMID- 9394587 TI - [Rupture of the papillary muscle after percutaneous transvenous mitral commissurotomy (PTMC)--a case report]. AB - We experienced a rare case of the mitral regurgitation due to papillary muscle rupture after percutaneous transvenous mitral commissurotomy (PTMC). This case was a seventy years old female who underwent PTMC. The cardiac tamponade and mitral regurgitation occurred after PTMC. Pericardial drainage was done immediately, and the next day the emergency operation was required. Rupture of the posterior papillary muscle was found at the operation, and mitral valve replacement was performed. Her postoperative course was uneventful and she discharged on the 26th day after the operation. We should take the papillary muscle rupture into consideration if there are severe sub-valvular lesion and shorting of the chorda. PMID- 9394588 TI - [Right-sided intrathoracic bypass grafting for a coarctation of the aorta in an advanced aged woman]. AB - A sixty-two years old woman with a successful bypass grafting for coarctation of the thoracic aorta (Co-A) is presented. She has been suffering from hypertension since her thirties. Preoperative cardiac catheterization demonstrated Co-A with a pressure gradient of 100 mmHg. Angiography revealed the hypoplastic aortic arch with marked calcification and well developed collateral circulation. Right-sided intrathoracic bypass grafting using a 16 mm woven Dacron graft was placed between the ascending and descending aorta via a right sixth intercostal space. The patient had a good postoperative course with reduced pressure gradient of 10 mmHg. We recommend this procedure to be a safe and simple technique that can avoid injury to the collateral circulation to our knowledge, this is the most aged patient who underwent a successful surgical treatment for the Co-A in Japan. PMID- 9394589 TI - [A case of ruptured thoracoabdominal aortic aneurysm with aortitis syndrome- operation with selective cold visceral arteries perfusion]. AB - We report a successful result of treatment for a ruptured thoracoabdominal aortic aneurysm with aortitis syndrome. A 43-year-old male suffered sudden low back pain, that was diagnosed as a ruptured thoracoabdominal aortic aneurysm based on abdominal computed tomography. Preoperative angiography revealed a thoracoabdominal aortic aneurysm with occlusion of the superior mesenteric artery, and well developed Riolan's archade. The aneurysm was replaced by a prosthetic graft with partial femoro-femoral bypass in conjunction with selective cold perfusion for the visceral arteries. Total extracorporeal circulation time, and aortic clamptime, was 187 minutes and 132 minutes, respectively. The postoperative courses of liver and renal function were excellent. The patient recovered from surgery uneventfully. It was suggested that selective cold visceral perfusion was effective for prevention of renal and liver dysfunction associated with a ruptured thoracoabdominal aneurysm. PMID- 9394590 TI - [Successful two-stage approach to treating excessive hemorrhage from pulmonary arterial stump in post-lobectomy bronchopleural fistula]. AB - A 62-year-old man underwent right lower lobectomy for adenocarcinoma (pT2N0M0) and nine days later, a bronchopleural fistula with empyema was evident. Six weeks following the lobectomy, excessive hemorrhage from the site of chest drainage and hemoptysis were noted. The bleeding and empyema were controlled by a two-stage approach. Anterior transpericardial approach was first made through the median sternotomy to clamp the right main pulmonary artery and then postero-lateral thoracotomy was conducted for the bronchopleural fistula with empyema. The right bronchial stump was covered with a pedicled muscle flap and pseudomonas aeruginosa, always positive in drainage effusion, consequently disappeared. The patient was discharged with a closed bronchus 4 months following the operation. PMID- 9394591 TI - [Successful emergency surgical management following cardiac massage in a patient with acute myocardial infarction due to total obstruction of the left main trunk]. AB - The prognosis in patients manifesting shock following acute myocardial infarction due to total occlusion of the left main trunk (LMT) is usually very poor and so is the lifesaving rate. Accurate judgement and rapid response are key to the successful management of this disease. We experienced a successful case with emergency coronary artery bypass grafting (CABG) on the 14 the day after initial attack. The patient, who had total occlusion of LMT, underwent a PTCA (percutaneous transluminal coronary angioplasty) during the initial attack under cardiac massage. We think in situations where patients have cardiac arrest, shock, elevated CPK levels suggesting devastation of myocardium due either to LMT or severe triple vessels disease, early catheter intervention rather than emergency CABG would be much more tolerable as long as hemodynamic situation allows. Our previous experience taught us that immediate surgical intervention with CABG usually resulted in poor outcome. Further refinements regarding the surgical procedure, technique, assist circulatory supports, cardioplegia, etc., are indispensable before trying to have a successful emergency CABG. PMID- 9394592 TI - [Repair of aortic arch aneurysm protruding to the retrobronchial space--a case report]. AB - A case of pseudoaneurysm of the aortic arch which protruded to the retrobronchial space is reported. A 73-year-old female complaining of severe chest pain was transferred to our hospital, CT showed an abnormal mass which occupied the retrobronchial space and displaced the esophagus toward righ, associated with left pleural effusion. A pseudoaneurysm of the aortic arch was suspected. Angiography revealed an aortic arch aneurysm protruding to the retrobronchial space. Emergent total arch replacement was performed. We diagnosed it as an impending rupture of the aortic arch aneurysm with a specific shape. PMID- 9394593 TI - [Successful surgical correction of incomplete endocardial cushion defect in a 65 year-old female]. AB - We report a case of surgical correction of a 65-year-old female. She presented severe congestive heart failure and preoperative cardiac catheterization showed massive left to right shunt (87%), mild mitral regurgitation, severe tricuspid regurgitation and pulmonary hypertension. The operative procedure consisted of annuloplasty of mitral valve (Kay's method), patch closure of the ostium primum defect and annuloplasty of tricuspid valve. Postoperative examination showed complete competence of mitral valve and improved functional capacity. This is the fourth successful case report of surgical correction of incomplete endocardial cushion defect in patients older than 65-year-old in Japan to our knowledge. Surgical correction of incomplete endocardial cushion defect should be recommended even in elder patients. PMID- 9394594 TI - [An surgical case of right pulmonary coccidioidomycosis--with subcutaneous coccidiomycosis in the left wrist]. AB - Coccidioides is an afferent fungus disease. In Japan, there have been only a few surgical reports on coccidioides disease. We report a 39-year-old male who was diagnosed as having coccidioides disease by biopsy of subcutaneous nodules in the left wrist. The patient also showed a tumor image (1.5 x 1.0 cm) in S4 in the right lung. He had previously lived in Fresno, California on business between 1988 and 1993. After biopsy of the subcutaneous nodules, Itraconazole (200 mg), an anti-fungal drug, was orally administered for the lesion in the right lung for about 6 months. Since the tumor image revealed no improvement through this treatment, the tumor was resected. Histopathological examination by Grocott staining demonstrated the spherical form Coccidioides, i.e., endospores. Only 5 cases of resected pulmonary coccidioidal lesions have been reported in Japan including this case. We must be careful when handling coccidioidal culture because of its strong infectiosity. PMID- 9394595 TI - [An adult case of aortic coarctation associated with two thoracic aneurysms]. AB - A 57-year-old woman in whom an abnormality was detected on the chest X-ray presented with no signs or symptoms other than hypertension. Several examinations revealed that she had aortic coarctation of the isthmus with two aneurysm in the arch. One aneurysm was located in the root of the left subclavian artery, another was just distal of the first aneurysm. For prevention of rupture of the aneurysms and treatment of hypertension, aortic arch reconstruction was performed with the aid of selective cerebral perfusion. The postoperative course was uneventful and she was discharged 19 days after the operation with normalization of her blood pressure. At the operation in this case, the combination of the two approaches, median sternotomy and left 4th thoracotomy, was useful. PMID- 9394596 TI - [Minimally invasive approach for mitral valve, aortic valve, and atrial septal defect surgery]. AB - We successfully introduced minimally invasive cardiac surgery (MICS) to japan by performing thoracoscopic clipping of a patent ductus arteriosus in July 1992. MICS via a small right parasternal incision (Cosgrove procedure) was applied for one patients with severe rheumatic mitral stenosis, one with severe aortic regurgitation, and one with atrial septal defect (ASD). Mitral valve replacement (MVR), aortic valve replacement (AVR), and direct closure of the ASD were performed successfully by MICS for the the first time in Japan. All three patients required no blood transfusion and had no complications postoperatively, being discharged from hospital at 15, 13, and 9 days after their operations. MICS was satisfactory for mitral valve and ASD operations, but AVR by this approach took much longer than by standard midline sternotomy due to the poor surgical field obtained via the small right parasternal incision. A minimally invasive approach for surgery on the aortic valve and ascending aorta may require transection of the sternum or some other method. MICS has several advantages, including less trauma and pain, faster patient recovery, shorter ICU and hospital stays, a lower cost, and a better cosmetic outcome. Therefore, it is better for the patient when it is feasible. MICS should develop and be applied to more patients with cardiovascular disease in the future. Some of the standard cardiovascular operations may soon be replaced by MICS. PMID- 9394597 TI - [A successful surgical treatment for huge left circumflex artery-right ventricle fistula using the fistula occlusion test]. AB - A surgical case of rare coronary artery fistula between the left circumflex artery and right ventricle was reported. A 46-year-old woman had suffered from exertional dyspnea and palpitation for three years. Pan systolic heart murmur was heard through the left 4th inter costal space. The chest X-ray film demonstrated cardiac enlargement and lung congestion, and the electrocardiograms showed atrial fibrillation and left ventricular hypertrophy when she was admitted to our hospital. Preoperative catheterization revealed a huge coronary artery fistula originating from the left circumflex artery and opening into the right ventricle through the posterior wall of the heart. The left-to-right shunt ratio was 60% and Qp/Qs was 2.47. At operation, the dilated circumflex artery fistula was carefully dissected and the tape was passed around the fistula as a tourniquet under extra corporeal circulation on the beating heart. To estimate myocardial ischemia, the fistula occlusion test was performed by tightening the previously placed tourniquet. Monitoring of electrocardiograms, transesophageal echocardiography, and hemodynamics were useful to detect myocardial ischemia. The occlusion test was performed under ECC for 5 minutes. No ischemic changes were observed. The fistula was interrupted under cardiac arrest at the point of the occlusion test. PMID- 9394598 TI - [Localized lesions on MRI in the globus pallidus, subthalamic nuclei and hippocampus in patients with severe intellectual and motor disabilities]. AB - We examined the clinical picture of eight patients with severe intellectual and motor disabilities, who had experienced prolonged and severe neonatal jaundice, and showed localized lesions in the globus pallidus, subthalamic nuclei and hippocampus on MRI. All patients had athetoid tetraplegia, and five patients showed disturbed ocular movements and seven showed dysphagia. Five patients could communicate with others or utter words, and all showed mental retardation. Auditory brainstem responses were abnormal in seven, and the percentage of REM sleep on all-night polysomnography was reduced in three. Neither CT nor T1 weighted MR images could detect any changes in the pallidum or subthalamic nuclei, while T2-weighted MR images disclosed bilateral high signals in the pallidum, especially in the internal segment, in all patients. Five of the 7 patients, in whom coronal T2-weighted MR imagings were obtained, showed high signals in the subthalamic nuclei. The hippocampus showed atrophy and/or T2 prolongation in seven patients. In one autopsy case, these MRI changes were concordant with pathological lesions. In patients with athetoid cerebral palsy, brainstem dysfunctions, and abnormal ABR, localization of MRI lesions to the pallidum and subthalamic nuclei is evidence for neonatal bilirubin encephalopathy. PMID- 9394599 TI - [Clinico-etiological study of forty-five cases of children with severe motor and intellectual disabilities of postnatal origin]. AB - The aim of this study is to analyze causes of severe brain damages of postnatal origin in children and to search for strategies to prevent them. The patients group consists of forty-five children with severe motor and intellectual disabilities sampled at several hospitals and special schools in a part of Tokyo. Twenty-four out of 45 cases (53%) were due to infectious diseases of the central nervous system (meningitis, encephalitis, and acute encephalopathy including Reye syndrome). Nine cases (20%) were due to brain damage related to medical services (complications of heart surgery, hypoglycemic encephalopathy, and so on). Accident-related brain damages accounted for 8 cases (18%) and 4 out of 8 were anoxic encephalopathy due to asphyxia (hanging and near drowning in two cases each). We conclude that intensive prevention and treatment of infectious diseases and accidents in children can reduce large part of the incidence of postnatally acquired severe brain damages in children. PMID- 9394600 TI - [Mismatch negativity of patients with hydranencephaly]. AB - We examined auditory evoked potentials and passive event-related potentials in two patients with hydranencephaly. In the middle latency response, a Na component was observed in both cases. Mismatch negativity was elicited in response to tone bursts and three patterns of vowel sounds in Patient 1, and three patterns of vowel stimuli in Patient 2. These results implicate the subcortical components of the auditory system in the generation of mismatch negativity. PMID- 9394601 TI - [Proinflammatory cytokine levels in cerebrospinal fluid from children with acute encephalitis]. AB - We investigated interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), and soluble TNF receptor 1 (sTNF-R 1) during the acute stage in the cerebrospinal fluid (CSF) from children with acute encephalitis by means of a sandwich enzyme immunoassay. We divided the 24 children with acute encephalitis into two groups: those who survived without neurological sequelae (Group 1, n = 15), and those who died or were left with sequelae (Group 2, n = 9). The IL-1 beta, IL-6, TNF-alpha and sTNF-R 1 levels in CSF in the two groups were significantly higher than those in control subjects (n = 23). The CSF sTNF-R 1 levels in Group 2 were significantly higher than those in Group 1. Our findings suggest that the IL-1 beta, IL-6 and TNF-alpha in CSF are related to the pathogenesis of acute encephalitis, and that the CSF level of sTNF R1 during the acute stage of encephalitis is an important index for predicting the neurological outcome. PMID- 9394602 TI - [Moyamoya disease presenting initially with mental disturbance]. AB - Moyamoya disease usually presents itself in children as recurrent episodes of transient cerebral ischemia, such as acute motor and sensory deficits, speech disturbance, headache and seizures. Its initial symptoms rarely include mental disturbance. We experienced a nine-year-old girl with Moyamoya disease who showed choreic involuntary movements of the left upper and lower limbs. In spite of mental disturbance which began around the age of four, she had never visited hospital. CT and MRI detected multiple ischemic lesions in the frontal, parietal and occipital areas. SPECT demonstrated low perfusion of these regions as well as of the right corpora striata, which appeared to be normal in MRI. She underwent right and left superficial temporal artery-middle cerebral artery (STA-MCA) anastomoses with an interval of three months. The choreic movement completely disappeared but her intelligence showed no improvement. The cerebral blood flow increased, but there was no change in the infarction area. Moyamoya disease should be considered in the differential diagnosis of mental disturbance in young children. PMID- 9394603 TI - [A case of pacemaker implantation for complete atrioventricular block associated with Duchenne muscular dystrophy]. AB - We report here a case of Duchenne muscular dystrophy (DMD) who underwent pacemaker implantation for complete atrioventricular block. This 30 year-old male had the deletion of exon 45-52 in the dystrophin gene and complained of palpitation and precordial oppression. Because his electrocardiogram showed complete atrioventricular block, a permanent pacemaker was implanted. He has been doing well for 15 months after implantation. There have been few reports about pacemaker implantation for patients with DMD. As these patients survive longer by mechanical ventilation with chest respirators or nasal intermittent positive pressure ventilators, an increasing number of cases may require pacemaker implantation. PMID- 9394604 TI - [A case of postencephalitic intractable partial epilepsy with multiple brain lesions demonstrated by MRI]. AB - We encountered a case of focal encephalitis. A 5-year-old boy developed high fever and he was admitted to our hospital on the third day with generalized tonic clonic convulsions. Cerebrospinal fluid showed slight pleocytosis. CT showed diffuse low-density area in the left temporal, parietal and occipital regions. T2 weighed MRI showed swelling and hyperintense regions in the left temporal, parietal and occipital cortex. With the disappearance of generalized convulsion, the cortical swelling improved. But aphasia and intractable complex partial seizures appeared. On MRI, the atrophic findings of the left hippocampus and temporal lesion became developed. We considered this case corresponds to focal encephalitis related to "a peculiar type of post-encephalitic/encephalopathic epilepsy" reported by Awaya et al. PMID- 9394605 TI - [A case of PEHO (progressive encephalopathy with edema, hypsarrhythmia and optic atrophy) syndrome: changes in clinical and neuroradiological findings]. AB - We reported serial clinical, radiological, and neurophysiological findings of a patient with PEHO (progressive encephalopathy with edema, hypsarrhythmia and optic atrophy) syndrome. The case was a 4-year-and-8-month-old boy. He had no apparent problems during pregnancy, but after his birth severe hypotonia and developmental delay were evident. Infantile spasms appeared at 3 months of age, and progressive opisthotonic posture and loss of his visual acuity at 8 months. Eye fixation was lost, and optic atrophy was observed. Seizures and progressive atrophy of the cerebellum and brainstem developed during the same period. These findings of our case support the hypothesis that brainstem lesions are related to infantile spasms. PMID- 9394606 TI - [Fulminant subacute sclerosing panencephalitis: clinical and neuropathological observations]. AB - We describe a 3-year-old boy with subacute sclerosing panencephalitis (SSPE) who died 4 months after its onset. His initial symptoms were drowsiness and left hemiplegia. He became comatose in 10 days, and developed a decortical posture after 45 days. He suffered from multiple cerebral hemorrhage and infarction 3 months later. Oligodendrocytes were positively stained by immunocytochemical stain with a complement-fixing measles antibody. Light microscopy revealed glial nodules, perivascular cuffing and reactive gliosis. Small arteries showed intimal thickening with resultant occlusion and occasional recanalization. These findings suggested vascular involvement in SSPE. This case illustrates the difference between the fulminant and chronic forms of SSPE. PMID- 9394607 TI - [A 10-year-old case with idiopathic oculomotor nerve palsy]. AB - We reported a 10-year-old girl with acquired left oculomotor nerve palsy. Neurologic and radiological examinations failed to reveal the etiology. Following administration of corticosteroid and vitamin B6, diplopia improved within 6 weeks, and mydriasis has been improving over the past 9 months. Idiopathic acquired oculomotor nerve palsy is a very rare condition in childhood, and prognosis of the disease is sometimes good. PMID- 9394608 TI - [Female autopsy case of Seitelberger's connatal form of Pelizaeus-Merzbacher disease]. AB - We report here an autopsy case of connatal Pelizaeus-Merzbacher disease, the second from Japan. Her clinico-pathological findings were essentially the same as those of the first case that we reported previously. The clinical course of this patient was 19 years in duration. Pathologically myelin had disappeared from the entire central nervous system, whereas that of the peripheral nervous system was preserved. Axons appeared also intact except for torpedo formation in the cerebellum. Demyelinated areas showed isomorphic gliosis. Recent studies have revealed impairment of proteolipid protein synthesis in classical Pelizaeus Merzbacher disease, the causative gene of which being located on the X chromosome. Thus, this disease is inherited as an X-linked recessive trait. However we report herein a sporadic female case in a non-consanguineous family. Therefore, we propose that this disease might have another causative gene and/or another mode of inheritance. PMID- 9394609 TI - [A case of floppy infant with diffuse cerebral calcification, atresia ani, mental retardation and epilepsy]. PMID- 9394610 TI - Histamine and eicosanoid levels in plasma and peripheral blood leucocyte cultures during experimental infection of cattle with bluetongue virus serotype 11. AB - Three calves were sensitized with three doses of inactivated BTV-11 UC8 strain and then experimentally infected with the homologous virus. In addition, four BTV seronegative heifers were also experimentally infected with BTV-11. Granulocyte rich fractions of peripheral blood leucocyte (PBL-GRF) cultures from BTV sensitized/infected calves and from control unexposed cattle were exposed in vitro with BTV-11. Histamine, leukotriene (LT) C4 and prostaglandin (PG) D2 were assayed in supernatant fluids. Plasma histamine levels increased in BTV-infected heifers from 10.1 +/- 2 ng/ml at Day 0 to 23.1 +/- 6.6 ng/ml at Day 12 following virus exposure. In addition, in this experimental group the concentration of PGF2 alpha (mean 551.97 +/- 243.54 pg/ml) increased significantly (P < or = 0.05) compared with control cattle (mean 467.3 +/- 73.9 pg/ml). Bluetongue virus induced histamine and LTC4 release after in vitro infection of PBL-GRF. Release of LTC4 was significantly (P < or = 0.05) higher in PBL-GRF cultures from sensitized and control animals than in unexposed PBL-GRF cultures. In contrast to these results, PGD2 was not released after BTV infection of PBL-GRF in vitro. The histamine release caused by BTV was virus-specific and mainly mediated by an immunological reaction, since the release was significantly reduced by removal of cell surface immunoglobulins. PMID- 9394611 TI - Experimental infection of swine and cat central nervous systems by the pig paramyxovirus of the blue eye disease. AB - This study analyses whether the pig paramyxovirus of blue eye disease (PPBED) infects the central nervous system (CNS) utilizing anterograde and retrograde peripheral nerve transport systems. The virus was injected into muscle and skin, and inoculated per nasum. The presence of PPBED was detected by an immunohistochemical method using polyclonal mouse antibodies against the whole inactivated virus, and was revealed with polyclonal rabbit antibodies against mouse immunoglobulin G (IgG) labelled with peroxidase. The PPBED injected into the pig medial gastrocnemius (MG) muscle was detected in a terminal branch innervating the MG muscle, in neural fibres of the sciatic nerve, in fibres of the ventral and dorsal spinal roots and in ventral horn neurones of the spinal cord. When PPBED was injected into the skin area innervated by the sural nerve, it was detected in neural fibres of the sural and sciatic nerves and in spinal cord dorsal horn neurones. The per nasum inoculum rapidly invaded the CNS through the olfactory nerve. The study concluded that, in order to invade the CNS, PPBED was transported retrogradely by peripheral cutaneous and muscular nerves, and anterogradely by the olfactory nerve. No PPBED was detected in either cat peripheral nerves or in cat CNS. PMID- 9394612 TI - Monoclonal antibody characterization of rabies virus strains isolated in the River Plate Basin. AB - In this study, 91 strains isolated in the River Plate Basin, South America, were examined from the epidemiological standpoint and with monoclonal antibodies (MAbs) to the nucleocapsid of rabies virus. Such strains reacted to MAbs in accordance with nine different patterns (antigenic variants). Rabies virus was isolated from 49 cattle, 21 dogs, 11 non-haematophagous bats, four vampire bats, two foxes, two horses, one buffalo, and one human. Five of the variants had not been described previously. It was also found that two cases of rabies in wild foxes (Cerdocyon thous) which had attacked persons in the Province of Chaco, Argentina, had been caused by variants from dog and vampire bat, while two cases in frugivorous bats (Artibeus lituratus) from Argentina and Brazil, had been infected by vampire bat variants. In addition, symptoms shown by cattle infected with strains which reacted as originating in canine vectors, differed from those observed in bovines from which the variants isolated corresponded to vampire bats. PMID- 9394613 TI - Immunopotentiating activities of polysaccharide fraction from pine seed shell extract. AB - The function of polysaccharide (PSE) extracted from pine seed shells as a immunopotentiator was investigated. The phagocytosis and chemotaxis of mouse peritoneal macrophages (MP) were augmented by oral administration of PSE. The incorporation of 3H-thymidine by Con A-stimulated mouse splenic cells was significantly intensified by administration of PSE but the adherent-cells (MP) eliminated splenic lymphocytes was not increased. It is shown that the existence of activated MP is needed for the activation (proliferation) of T lymphocytes. The responsiveness of mouse T cells to alloantigen was also augmented by administration of PSE. The number of anti-SRBC plaque-forming cells in the spleen of mouse and chicken was also significantly increased. The results indicate that MP are first activated by oral administration of PSE and that the activation of T cells is then initiated indirectly by humoral factors (cytokines) produced by activated MP. PMID- 9394614 TI - Partial purification and characterization of a murine malaria parasite, Plasmodium berghei specific aldolase. AB - Cell-free P. berghei contains 26.1 times more aldolase activity as compared to normal mouse erythrocytes. Subcellular fractionation of cell-free parasite showed maximum enzyme activity in the soluble fraction. The parasite enzyme was active in a narrow pH range of 7.8-8.0. Of the enzyme activity 90% was lost within 2 weeks at 4 degrees C. Slight inhibition was observed with specific inhibitors ATP, pyrophosphate (PPi) and PEP. The F1, 6DP Km was 0.025 mM. PMID- 9394615 TI - Intestinal manifestations of experimental SIV-infection in rhesus monkeys (Macaca mulatta): a histological and ultrastructural study. AB - Intestinal lesions were studied in 32 rhesus monkeys experimentally infected with different strains of simian immunodeficiency virus SIVmac (251/32H, 251/32H-SPL and 251/MPBL) by light microscopy, transmission and scanning electron microscopy. A spectrum of primary and secondary manifestations of SIV-infection were detected. Primary changes included 'SIV-enteropathy' in 12 monkeys and virus induced syncytial giant cell formation (GCF) of the intestine in two animals. A primary virus-induced enteropathy occurred both as only histologically visible 'SIV-enteropathy' and as 'AIDS-enteropathy' accompanied by clinical signs of enteritis. Secondary opportunistic infections (Balantidium coli, Cryptosporidium, Trichuris, Trichomonas, Spironucleus, Mycobacteria and Cytomegalovirus) were identified in 27 animals and three monkeys developed malignant lymphomas involving the intestinal tract. Compared to intestinal lesions in HIV-infected patients, differences were found concerning the incidence of GCF and the range of opportunistic infections, with cryptosporidium, cytomegalovirus and mycobacteria occurring in both SIV-infected macaques and AIDS patients. The present observations revealed that SIV-infected rhesus monkeys provide an excellent model both for studies on the pathogenesis of HIV-enteropathy and opportunistic infections and for the development of therapies against cryptosporidial, cytomegalovirus and mycobacteria infection. Comparison of three SIV-strains revealed differences in primary and secondary lesions observed: SIVmac251/MPBL was correlated with severe primary SIV-induced pathologic changes and SIVmac251 SPL-infected animals showed a higher incidence of malignant lymphomas. PMID- 9394616 TI - The chromosomal signal for sex determination in Caenorhabditis elegans. AB - In Caenorhabditis elegans, sex is determined by the number of X chromosomes which, in turn, determines the expression of the X-linked gene xol-1. Recent work has shown that xol-1 expression is controlled by least two distinct regulatory mechanisms, one transcriptional and another post-transcriptional. The transcriptional regulator is a repressor acting in XX embryos; although the specific gene has not been identified, the chromosome region has been defined. A previously defined regulator of xol-1, known as fox-1, maps to a different region of the X chromosome and works post-transcriptionally, consistent with the identification of the fox-1 gene product as a putative RNA binding protein. PMID- 9394617 TI - Zyxin: zinc fingers at sites of cell adhesion. AB - Zyxin is a low abundance phosphoprotein that is localized at sites of cell substratum adhesion in fibroblasts. Zyxin displays the architectural features of an intracellular signal transducer. The protein exhibits an extensive proline rich domain, a nuclear export signal and three copies of the LIM motif, a double zinc-finger domain found in many proteins that play central roles in regulation of cell differentiation. Zyxin interacts with alpha-actinin, members of the cysteine-rich protein (CRP) family, proteins that display Src homology 3 (SH3) domains and Ena/VASP family members. Zyxin and its partners have been implicated in the spatial control of actin filament assembly as well as in pathways important for cell differentiation. Based on its repertoire of binding partners and its behavior, zyxin may serve as a scaffold for the assembly of multimeric protein machines that function in the nucleus and at sites of cell adhesion. PMID- 9394618 TI - Root development: signaling down and around. AB - Because of its elegant simplicity, the Arabidopsis root has become a model for studying plant organogenesis. In this review we focus on recent results indicating the importance of signaling in root development. A role for positional information in root cell specification has been demonstrated by ablation analyses. Through mutational analysis, genes have been identified that play a role in radial pattern formation. The embryonic phenotypes of these mutants raised the possibility that division patterns in post-embryonic roots are dependent on signaling that originates during embryonic development. Analysis of expression of the SCARECROW gene indicates that it may play a role in this 'top down' signaling process. Characterization of root epidermis development has led to the identification of negative regulators of root-hair formation. These appear to set up a prepattern which is reinforced by signaling by plant hormones. PMID- 9394619 TI - Proliferating cell nuclear antigen: more than a clamp for DNA polymerases. AB - DNA metabolic events such as replication, repair and recombination require the concerted action of several enzymes and cofactors. Nature has provided a set of proteins that support DNA polymerases in performing processive, accurate and rapid DNA synthesis. Two of them, the proliferating cell nuclear antigen and its adapter protein replication factor C, cooperate to form a moving platform that was initially thought of only as an anchor point for DNA polymerases delta and epsilon. It now appears that proliferating cell nuclear antigen is also a communication point between a variety of important cellular processes including cell cycle control, DNA replication, nucleotide excision repair, post-replication mismatch repair, base excision repair and at least one apoptotic pathway. The dynamic movement of proliferating cell nuclear antigen on and off the DNA renders this protein an ideal communicator for a variety of proteins that are essential for DNA metabolic events in eukaryotic cells. PMID- 9394620 TI - Multifunctional plasma membrane redox systems. AB - All the biological membranes contain oxidoreduction systems actively involved in their bioenergetics. Plasma membrane redox systems seem to be ubiquitous and they have been related to several important functions, including not only their role in cell bioenergetics, but also in cell defense through the generation of reactive oxygen species, in iron uptake, in the control of cell growth and proliferation and in signal transduction. In the last few years, an increasing number of mechanistic and molecular studies have deeply widened our knowledge on the function of these plasma membrane redox systems. The aim of this review is to summarize what is currently known about the components and physiological roles of these systems. PMID- 9394621 TI - Budding of enveloped viruses from the plasma membrane. AB - Many enveloped viruses are released from infected cells by maturing and budding at the plasma membrane. During this process, viral core components are incorporated into membrane vesicles that contain viral transmembrane proteins, termed 'spike' proteins. For many years these spike proteins, which are required for infectivity, were believed to be incorporated into virions via a direct interaction between their cytoplasmic domains and viral core components. More recent evidence shows that, while such direct interactions drive budding of alphaviruses, this may not be the case for negative strand RNA viruses and retroviruses. These viruses can bud particles in the absence of spike proteins, using only viral core components to drive the process. In some cases the spike proteins, without the viral core, can be released as virus-like particles. Optimal budding and release may, therefore, depend on a 'push-and-pull' concerted action of core and spike, where oligomerization of both components plays a crucial role. PMID- 9394622 TI - Signaling activation and repression of RNA polymerase II transcription in yeast. AB - Activators of RNA polymerase II transcription possess distinct and separable DNA binding and transcriptional activation domains. They are thought to function by binding to specific sites on DNA and interacting with proteins (transcription factors) binding near to the transcriptional start site of a gene. The ability of these proteins to activate transcription is a highly regulated process, with activation only occurring under specific conditions to ensure proper timing and levels of target gene expression. Such regulation modulates the ability of transcription factors either to bind DNA or to interact with the transcriptional machinery. Here we discuss recent advances in our understanding of these mechanisms of transcriptional regulation in yeast. PMID- 9394623 TI - Calpains: intact and active? AB - Calpains are a family of calcium-dependent thiol-proteases which are proposed to be involved in many physiological processes as well as pathological conditions. Calpains are likely to be involved in processing of numerous enzymes and cytoskeletal components, thereby linking their activity to a variety of intracellular events. Although widely studied, the precise mechanism(s) involved in calpain activation and activity in vivo remain poorly understood. Initial studies suggested that calpain exists primarily as an inactive proenzyme that required autolytic cleavage for activation. It was also hypothesized that calpain associated with membrane phospholipids, serving to increase calcium sensitivity, facilitating autolytic conversion and thus activating the enzyme. These hypotheses, however, have not been universally accepted and there is increasing evidence that intact, non-autolyzed calpain is the physiologically active calpain form. PMID- 9394624 TI - Antimicrobial peptide defense in Drosophila. AB - Drosophila responds to a septic injury by the rapid synthesis of antimicrobial peptides. These molecules are predominantly produced by the fat body, a functional equivalent of mammalian liver, and are secreted into the hemolymph where their concentrations can reach up to 100 microM. Six distinct antibacterial peptides (plus isoforms) and one antifungal peptide have been characterized in Drosophila and their genes cloned. The induction of the gene encoding the antifungal peptide relies on the spatzle/Toll/cactus gene cassette, which is involved in the control of dorsoventral patterning in the embryo, and shows interesting structural and functional similarities with cytokine-induced activation of NF-kappa B in mammalian cells. An additional pathway, dependent on the as yet unidentified imd (for immune-deficiency) gene, is required for the full induction of the antibacterial peptide genes. Mutants deficient for the Toll and imd pathways exhibit a severely reduced survival to fungal and bacterial infections, respectively. Recent data on the molecular mechanisms underlying recognition of non-self are also discussed in this review. PMID- 9394625 TI - Does sex determination start at conception? AB - Recent molecular studies of mammalian sexual determination have been focused on gene expression in the gonadal ridge at the time of appearance of sexual dimorphism: the critical time defined by the 'Jost principle'. Three lines of evidence suggest that, instead, sex determination may start shortly after conception: (1) the XY preimplantation embryo usually develops more rapidly than the XX preimplantation embryo (this phenotype has been linked to the Y chromosome and will be termed 'Growth factor Y'); (2) the gene for testis determination, SRY/Sry, and the closely linked genes ZFY/Zfy and Smcy, are transcribed in the preimplantation embryo; and (3) male and female preimplantation embryos are antigenically distinguishable, indicating sex differences in gene expression. The data to support these assertions are reviewed. Possible relationships of these three phenomena to each other and to sex differentiation are discussed. Similarities in mechanisms of sexual determination between marsupial and eutherian mammals are hypothesized. Problems with interpreting male sexual differentiation as being solely due to testosterone and Mullerian inhibiting substance (MIS) are discussed. PMID- 9394626 TI - Plastids in parasites of humans. AB - It has recently emerged that malarial, toxoplasmodial and related parasites contain a vestigial plastid (the organelle in which photosynthesis occurs in plants and algae). The function of the plastid in these obligate intracellular parasites has not been established. It seems likely that modern apicomplexans derive from photosynthetic predecessors, which perhaps formed associations with protists and invertebrates and abandoned autotrophy in favour of parasitism. Recognition of a third genetic compartment in these parasites proffers alternative strategies for combating a host of important human and animal diseases. It also poses some fascinating questions about the evolutionary biology of this important group of pathogens. PMID- 9394627 TI - Evolving rodent dentition. PMID- 9394628 TI - Cross-modal equivalence of visual and auditory scatterplots for exploring bivariate data samples. AB - The equivalence of visual and auditory scatterplots was examined in two experiments. Experiment 1 examined the relationship between actual Pearson's r and visual and auditory judgments of direction and magnitude of correlation for 24 bivariate data samples. Experiment 2 directly evaluated visual and auditory perceptual sensitivity to outliers by examining changes in perceived magnitude and direction of correlation estimates for scatterplots from Experiment 1 that were altered by the addition of outlier points. Results suggest that the information conveyed by visual and auditory scatterplots is used very similarly by the two modalities. Both visual and auditory scatterplots are quite efficient in conveying sign and magnitude of correlation, and the effect of outliers on judged magnitude of correlation is similar for the two types of data display. PMID- 9394629 TI - Artificial looming yields improved performance over lateral motion: implications for stereoscopic display techniques. AB - In the natural world, a number of visual cues indicate that an item is quickly approaching the perceiver. Binocular disparity is one cue for depth, and it has been demonstrated that abrupt changes in disparity, artificially unaccompanied by correlated depth cues, are capable of causing the perception of looming for the observer. An experiment involving 38 undergraduates, using a computer-controlled stereoscopic display, examined the ability of above-threshold changes in disparity (artificial looming) to facilitate response time and accuracy for observers engaged in an object-enumeration task within a cluttered display. Compared with performance using the same stimuli without disparity information (lateral motion), participants were more accurate regardless of the disparity level (9, 12, 24, or 48 minutes of arc) and faster at the two lowest levels of disparity. Participants showed the classic subitizing function, suggesting that target stimuli presented with motion information were segregated from otherwise identical distractor items. It is proposed that binocular disparity information can act as a valid location cuing method in stereoscopic computer displays in which form and color information are to be preserved. PMID- 9394630 TI - The effects of spatial frequency on binocular fusion: from elementary to complex images. AB - Factors limiting binocular fusion were studied using 2-dimensional difference of Gaussian (2D-DOG) stimuli. The proportion of fused stimuli and observer's response time were determined for stimuli that varied in spatial frequency composition between 0.22 and 4.8 cycles per degree. At small disparities, mean fusion response times were short and relatively stable but increased rapidly once the disparity reached a certain critical value. This 2-phase function implies the existence of 2 separate fusional mechanisms: a rapid neurally based fusional process, which operates at small disparities, and a second mechanism involving reflexive vergence movements operating at disparities 2 to 3 times larger. Both mechanisms are highly influenced by spatial frequency, being 4 to 5 times more effective at low spatial frequencies. Additional experiments demonstrated that with compound stimuli, the fusion limit is not determined by the highest spatial frequency components (as had been reported previously) but, rather, can take advantage of the additional fusional range associated with low spatial frequencies. Such cooperation may be obvious only in the case of 2-dimensional stimuli. PMID- 9394631 TI - Visual accommodation and virtual images: do attentional factors mediate the interacting effects of perceived distance, mental workload, and stimulus presentation modality? AB - A study was conducted to examine the effect on the visual accommodation response of processing information presented either visually or aurally while viewing a real-world scene. A number of different conditions were tested in which a mental processing task, a virtual image overlaying the real world (as is the case with head-up displays), or both were present. These manipulations provided two factors that have previously been demonstrated to have some influence on the accommodation response: mental effort and perceived distance. They also allowed the investigation of the influence of the source modality of the information to be processed. The results suggested that these influences may have a cumulative effect on the visual accommodation response, possibly mediated by attentional factors. PMID- 9394632 TI - Multiaccommodative stimuli in VR systems: problems & solutions. AB - Virtual reality environments can introduce multiple and sometimes conflicting accommodative stimuli. For instance, with the high-powered lenses commonly used in head-mounted displays, small discrepancies in screen lens placement, caused by manufacturer error or user adjustment focus error, can change the focal depths of the image by a couple of diopters. This can introduce a binocular accommodative stimulus or, if the displacement between the two screens is unequal, an unequal (anisometropic) accommodative stimulus for the two eyes. Systems that allow simultaneous viewing of virtual and real images can also introduce a conflict in accommodative stimuli: When real and virtual images are at different focal planes, both cannot be in focus at the same time, though they may appear to be in similar locations in space. In this paper four unique designs are described that minimize the range of accommodative stimuli and maximize the visual system's ability to cope efficiently with the focus conflicts that remain: pinhole optics, monocular lens addition combined with aniso-accommodation, chromatic bifocal, and bifocal lens system. The advantages and disadvantages of each design are described and recommendation for design choice is given after consideration of the end use of the virtual reality system (e.g., low or high end, entertainment, technical, or medical use). The appropriate design modifications should allow greater user comfort and better performance. PMID- 9394633 TI - A methodology for optimally designing console panels for use by a single operator. AB - A seven-step methodology is presented to determine a dimensionally correct optimal layout of a console panel for a single operator. This methodology integrates the steps in the layout design process and uses a mathematical optimization model from facility design to obtain the optimal panel layout. A major difference in this methodology from previous work is that the mathematical optimization model incorporates factors that are only partially included in previous mathematical models. In addition, it includes the areas of the panel components as a new factor. This methodology is illustrated by the design of a nuclear power plant console panel. PMID- 9394634 TI - Competition and performance on a computer-based complex perceptual-motor task. AB - Employees of temporary agencies practiced Space Fortress, a complex video game task, for 10 sessions, each consisting of 8 practice and 2 test games of 3 min each. Trainees practiced individually, in dyads, or in tetrads, and they were classified as having high or low aptitude based on computer attitude scores and baseline performance. Competition for monetary prizes was introduced early in training, late in training, or not at all. Competition facilitated high-aptitude trainees but not low-aptitude trainees. Group size and the timing of competition instructions had no main effects or interactions. The results are discussed in terms of social facilitation theory, according to which competition facilitates dominant responses, which helps high-skill trainees but not low-skill trainees. PMID- 9394635 TI - Framing of task performance strategies: effects on performance in a multiattribute dynamic decision making environment. AB - It is well documented that the way a static choice task is "framed" can dramatically alter choice behavior, often leading to observable preference reversals. This framing effect appears to result from perceived changes in the nature or location of a person's initial reference point, but it is not clear how framing effects might generalize to performance on dynamic decision making tasks that are characterized by high workload, time constraints, risk, or stress. A study was conducted to examine the hypothesis that framing can introduce affective components to the decision making process and can influence, either favorably (positive frame) or adversely (negative frame), the implementation and use of decision making strategies in dynamic high-workload environments. Results indicated that negative frame participants were significantly impaired in developing and employing a simple optimal decision strategy relative to a positive frame group. Discussion focuses on implications of these results for models of dynamic decision making. PMID- 9394636 TI - Aging and decision making: driving-related problem solving. AB - We examined age-related effects on decision making in a task environment familiar to most younger and older adults. Participants made route-selection decisions in real time. Participants received information about traffic density and expected speed limits of main and alternative routes, from which they determined the optimality of their present route versus alternative routes. The experiment evaluated the effects of information type, amount of congestion, alternative route speed limit, and age on speed and quality of decision making. Measures of optimal route selection revealed main effects of alternative route speed limit, congestion level, and message type, but there was not a main effect of age, and age did not interact with any variable. In terms of decision speed (but not quality of decision making), older participants were slower, and age interacted with alternative route speed and with message type. The data are interpreted in relation to previous data examining everyday problem solving and aging. PMID- 9394638 TI - Stability limits in extreme postures: effects of load positioning, foot placement, and strength. AB - Although injuries related to postural stability are prevalent, ergonomic job analyses traditionally have not addressed stability issues. In this research functional stability limits are quantified for persons standing in extreme postures under various external load and foot positioning conditions. Participants were asked to lean and displace their center of gravity (COG) as far as possible in eight directions to the sides and front of the body. Stability measures based on these COG displacements were calculated. All controlled variables significantly affected the stability measures. When standing unladen, participants extended their COG to within 99% of their theoretical maximum. Movement was much more restricted when handling a load (89%), especially when holding it with one hand on the shoulder (84%). On average, increased separation of the feet in a particular direction resulted in larger COG displacements in that direction. The results are discussed relative to their effects on balance and stability modeling. PMID- 9394637 TI - The influence of flooring on standing balance among older persons. AB - This study investigated the effects of flooring on balance during quiet standing in healthy young and older participants. Seven flooring conditions were examined, including a hard tile floor and combinations of low-pile and high-pile carpet with urethane foam and rubber padding. The resulting floors provided a variety of compliant surfaces, ranging from very soft to hard. Participants stood during three separate visual conditions: eyes open, eyes closed, and looking at a moving visual surround. Three measures of postural sway were calculated using center of pressure recordings during the trails. The results showed that the amplitude of sway was higher in the older than in the younger participants, particularly in the moving visual surround condition. Flooring compliance was found to have an effect on sway during moving visual environments, with the largest effects found among the older participants. Softer floors increased the amount of sway in the older participants. These results suggest that floor compliance influences standing postural stability in older people, particularly in destabilizing visual environments. PMID- 9394639 TI - Evaluation of the probability of spinal damage caused by sustained cyclic compression loading. AB - A sensitivity analysis of two alternative models predicting damage to vertebral motion segments (VMS) in cyclic compression was performed to evaluate the relative probability of damage occurring when peak compression force, loading frequency, or duration in a lifting task is changed. The first model is based on the assumption that fatigue failure is the mechanism underlying damage to the VMS in cyclic compression. The second model is based on the assumption that the VMS damage in cyclic compression is determined by the viscoelastic deformation of the segment and that the instant of failure can be predicted on the basis of the energy stored in this process. With both models, we estimated the percentage of the population likely to incur a VMS injury when performing a repetitive lifting task with peak compression forces ranging from 1500 to 4100 N, frequencies from 2 to 12 min-1, and durations between 30 and 120 min. The results indicate a dominant influence of the peak compression force on this outcome over the domain studied. This conclusion holds qualitatively for both models, suggesting that for a comparative analysis they can be considered interchangeable. However, a considerable quantitative difference in the absolute outcomes of the two models was found, which stresses the importance of further study on the validity of these models. PMID- 9394640 TI - Effects of team size on the maximum weight bar lifting strength of military personnel. AB - Teamwork is an essential element in the majority of critical Army lifting tasks. Therefore, an understanding of the relationship between individual and team lifting capacity is of great tactical importance. Twenty-three male and 17 female U.S. Army soldiers were randomly assigned to single- and mixed-gender teams of two, three, and four persons. Individual lifting strength was the one-repetition maximum (1RM) load lifted from floor to knuckle height using a weight bar. A square-shaped bar was used for two- and four-person lifting, and a triangular shaped bar was used for three-person lifting. Team lifting strength as a percentage of the sum of individual lifting strength (%sum) did not change with team size. The %sum for teams of men (87.3%) was less than for teams of women (91.1%, p < 0.05). The %sums for both single-gender teams (all men and all women) were greater (p < 0.01) than for mixed-gender teams (80.2%). The number of people lifting a large object was increased to four with no decrease in the effectiveness of the individual lifter beyond that found for two persons. The 1RM loads presented in this paper were lifted under ideal conditions by young soldiers and do not represent norms for an industrial population. PMID- 9394641 TI - The effect of valve handwheel type, operating plane, and grasping posture on peak torque strength of young men and women. AB - An experiment was designed to assess the factors affecting the operation of valve handwheels. Forty volunteers (20 men and 20 women) participated in this study. A nested-factorial experimental design was employed. Handwheel type (smooth, curved, or knurled rim), operating plane (sagittal, frontal, or transverse plane), grasping posture (power or precision grasp), and operating direction (clockwise or counterclockwise) were found to have significant effects on the (maximum volitional torque exertion [MVTE]). A power grasp exerted more force than did a precision grasp. A smooth-rim handwheel oriented in the frontal plane resulted in the least MVTE. Counterclockwise torque exertion was significantly greater than clockwise torque exertion, but the difference was not very large. MVTE for women (7.9 Nm) was about 66% of that for men (12.0 Nm). PMID- 9394642 TI - A dynamic mechanical model for hand force in right angle nutrunner operation. AB - A deterministic mechanical model based on physical tool parameters was used for estimating static and dynamic hand forces from kinematic measurements. We investigated the effects of target torque (25, 40, and 55 Nm) and threaded fastener joint hardness (35-, 150-, 300-, 500-, and 900-ms torque buildup time) on hand force. Estimated hand force was affected by target torque and joint hardness. Peak and average dynamic hand force was least for the hard joint (35-ms buildup) and greatest for the medium hardness joint (150-ms buildup). Tool inertia played the major role in reducing hand reaction force. Estimated hand force decreased when the inertial force component increased. Inertial force decreased by 366% when buildup time increased from 35 to 300 ms. Static modeling overestimated hand force; the error ranged from 10% for a soft joint to 40% for a hard joint. Results from direct hand force measurements using a strain gauge dynamometer showed that the dynamic model overestimated peak hand force by 9%. However, average hand force and force impulse were not significantly overestimated. PMID- 9394643 TI - Health risk assessment of fluctuating concentrations using lognormal models. AB - A mathematical model is proposed for assessing health risk rates of fluctuating concentrations. Each time-averaged concentration may be regarded as a dose that, when applied to the dose-response curve, produces a risk of an adverse effect. A theoretical derivation shows that the dose-response pattern is a cumulative lognormal curve because of the diversity of the individuals in the exposed population. Similarly, the concentration pattern is a log-normal distribution because of the diversity of emission sources and dispersive processes. The health risk is produced by the overlapping of the right tail of the concentration distribution and the left tail of the dose-response curve. The evaluation of the joint probability in this region has been performed by numerical integration by computer in terms of two generalized parameters. One represents the geometric standard deviation of the concentration distribution relative to that of the dose response curve, and the other represents the distance between the geometric mean concentration and the concentration producing an adverse response in 50% of the exposed population. These results are presented graphically and in tabular form. If the two parameters of the dose-response curve are known, the health risk of the concentration pattern may be calculated conveniently for any geometric mean and geometric standard deviation values. PMID- 9394644 TI - The origin and development of diagnostic radiology as illustrated by postage stamps. AB - This is a brief account of the scientific history of radiology and its use in medical imaging. The approach is untraditional as the narrative is highlighted with reproductions of selected postage stamps. These illustrations add a new dimension to the presentation of important events leading to the discovery and development of diagnostic radiology. PMID- 9394645 TI - The origin of the word Stent. AB - In 1856, the English dentist Charles Stent developed a thermoplastic-like material for taking impressions of toothless mouths. This "Stent mass" was later used as a device or mould for keeping a skin graft in place; it was also used to provide support for anastomosis. A hundred years after the inventor's death in 1885, the word stent has been adopted all over the world in interventional radiology but today it is understood to mean percutaneous tubular structures that induce or maintain lumen patency. The true origin of the word stent is not found in many dictionaries. In most references, the wrong dentist is given credit for the discovery. Dictionaries also refer to the obsolete English and Scottish words stent and stint which mean, among other things, "to extend". The true origin of the word is therefore somewhat unclear. PMID- 9394646 TI - MR imaging of the central nervous system in diving-related decompression illness. AB - PURPOSE: This investigation was conducted to determine whether MR imaging showed cerebral or spinal damage in acute diving-related decompression illness, a term that includes decompression sickness (DCS) and arterial gas embolism (AGE). MATERIAL AND METHODS: A total of 16 divers with dysbaric injuries were examined after the initiation of therapeutic recompression. Their injuries comprised: neurological DCS II n = 8; AGE n = 7; combined cerebral-AGE/spinal-DCS n = 1. T1- and T2-weighted images of the brain were obtained in 2 planes. In addition, the spinal cord was imaged in 7 subjects. The imaging findings were correlated with the neurological symptoms. RESULTS: MR images of the head showed ischemic cerebrovascular lesions in 6/8 patients with AGE but showed focal hyperintensities in only 2/8 divers with DCS. Spinal cord involvement was detected in 1/7 examinations, which was the combined cerebral-AGE/spinal-DCS case. There was agreement between the locations of the documented lesions and the clinical manifestations. CONCLUSION: MR readily detects cerebral damage in AGE but yields low sensitivity in DCS. A negative MR investigation cannot rule out AGE or DCS. However, MR is useful in the examination of patients with decompression illness. PMID- 9394648 TI - Proton (1H) MR spectroscopy for routine diagnostic evaluation of brain lesions. AB - PURPOSE: To describe the introduction and performance of proton MR spectroscopy (1H-MRS) in the daily routine of a modern standard MR unit. MATERIAL AND METHODS: Over an 8-month period, 52 patients with brain lesions were studied with 1H-MRS, using SE and STEAM sequences for chemical-shift imaging and single-volume spectroscopy. The quality of the spectra was graded from 1 (best) to 3, and the main factors influencing the quality of the spectra were evaluated. RESULTS: Of the measurements: 85% were graded as 1; 12% as 2; and 3% as 3. The main reasons for poor spectral quality were: the unfortunate positioning of the VOI; hemorrhage; and/or postoperative changes within the VOI. Of 40 patients with a final diagnosis: MRS provided an increased confidence in MR diagnosis in 18 cases; MRS contributed significantly to preoperative diagnosis in 3 cases; and the spectra were not specific (n = 10) or were difficult to evaluate (n = 9) owing to reduced quality (grade 2 or 3) in 19 cases. CONCLUSION: MRS of the brain can provide a high percentage of interpretable spectra and frequently can increase confidence in the MR diagnosis of brain lesions in clinical routine. PMID- 9394647 TI - Supratentorial primary intra-axial tumors in children. MR and CT evaluation. AB - PURPOSE: To evaluate the MR and CT features of pediatric supratentorial intra axial tumors with respect to differential diagnosis and the role of each investigation modality. MATERIAL AND METHODS: MR and CT findings in 40 children with 12 types of pathologically proven histological tumors were reviewed. RESULTS: The location of tumors might be one clue to differential diagnosis. In our material, cysts (60%), calcifications (45%), and intratumoral hemorrhages (27%) were found in the tumors. Characteristic features noted in some lesions included: peritumoral hemosiderin deposition in cavernous angiomas; intratumoral flow void in a choroid plexus carcinoma and in glioblastomas; and hemicerebral atrophy in germinomas. A comparison between malignant and benign tumors showed perifocal edema and a mass effect to be significantly more common in malignant lesions. Homogeneous enhancement suggested a benign tumor and an inhomogeneous pattern represented malignancy, while the lack of obvious enhancement did not always suggest benignity. Intratumoral calcium deposition was a not uncommon finding in malignant tumors. CONCLUSION: In most cases, the exact diagnosis should be made by histological examination but it is important for treatment planning that the appropriate depiction of tumor extension and tissue characterization be made by MR and CT. PMID- 9394649 TI - The perfusion fraction in volunteers and in patients with ischaemic stroke. AB - The fractional volume of capillary blood, i.e. the perfusion fraction f, was measured with the aid of an echo-planar imaging protocol originally designed for the measurement of water diffusion. In healthy volunteers, reasonable f values were obtained. In patients with cerebral ischaemic stroke, a marked decrease in the f value was seen in the infarcted region as compared with corresponding values in the contralateral hemisphere. We suggest that perfusion-fraction measurements may add to the diagnostic value of water-mobility examinations in patients with ischaemic disease. PMID- 9394650 TI - Detection of normal mediastinal lymph nodes by ultrasonography. AB - PURPOSE: The detection by US (in contrast to CT) of lymph nodes of any size in the mediastinum is usually considered to be a pathological finding. The aim of this study was to find out whether it was possible to detect normal lymph nodes by high-resolution mediastinal US. MATERIAL AND METHODS: Six different mediastinal regions in 80 healthy asymptomatic volunteers and in 20 human cadavers were examined by means of US (with colour Doppler imaging) to assess US access to the respective regions and to demonstrate the number and size of detectable lymph nodes. All the cadaveric lymph nodes that were detected were examined histologically to exclude inflammatory or malignant infiltration. RESULTS: In almost all subjects, we obtained US access to the supra-aortic (100%), paratracheal (95%), prevascular (99%), and pericardial (98%) regions, and to the aorticopulmonary window (98%). US access to the subcarinal region was more difficult (75%). In the healthy subjects, lymph nodes were detected in the paratracheal region (in 35% of these subjects, mean lymph-node diameter 12 x 7 mm), in the aorticopulmonary window (45%, 14 x 8 mm), and in the subcarinal region (13%, 13 x 7 mm). In the cadavers, histologically normal lymph nodes were detected frequently in the paratracheal region (85%, mean size 11 x 6 mm) and in the aorticopulmonary window (90%, 11 x 5 mm). CONCLUSION: These results indicate that normal lymph nodes (and not only pathological lymph nodes) can be demonstrated by high-resolution mediastinal US. PMID- 9394651 TI - Abdominal CT features and survival in acquired immunodeficiency. AB - PURPOSE: HIV-infected patients show a high incidence of abdominal disease. This investigation was made to determine whether abdominal CT provided prognostically relevant information in these patients. MATERIAL AND METHODS: Images from 533 abdominal CT examinations in 339 HIV-infected patients were retrospectively reviewed for signs of abdominal disease, and correlated with clinical data and survival rates. The Kaplan-Meier analysis and rank testing of survival, and proportional hazards regression were used to define prognostic clinical and imaging findings. RESULTS: Of the 339 patients, 278 (82%) showed abnormal abdominal findings on CT. Median survival was 29 months. Of the imaging findings, hepatic masses (n = 11), pathologically enlarged lymph nodes (n = 48), and ascites (n = 7) were associated with poor survival, giving a median survival of respectively 13 months, 15 months, and less than 1 month. These three features showed no association with CD4(+)-T-lymphocyte count or CDC category. Main determinants of survival were a low CD4(+)-T-lymphocyte count, and certain abnormal CT findings. Splenomegaly (n = 147), hepatomegaly (n = 144), and lymphadenopathy (n = 111) were the most common abdominal findings on CT but lacked prognostic relevance. CONCLUSION: Abdominal CT offered prognostic implications in HIV-infected patients and might serve in risk stratification in selected patients. CT features such as hepatic masses, grossly enlarged lymph nodes, or ascites indicate advanced immunosuppression. PMID- 9394652 TI - CT-guided core-needle biopsy in omental pathology. AB - PURPOSE: To assess the accuracy and clinical usefulness of CT-guided core-needle biopsy in the diagnosis of omental pathology. MATERIAL AND METHODS: We retrospectively reviewed the results of CT-guided percutaneous core biopsies in 25 patients with focal (n = 2) or diffuse (n = 23) omental pathology. These results were compared to the final diagnoses as determined by laparotomy (n = 15), laparoscopic biopsy (n = 3), endoscopic biopsy (n = 1), or by the results of percutaneous biopsy and clinical-radiological and bacteriological modalities (n = 6). The final diagnoses showed 4 patients with isolated omental pathology and 21 with widespread peritoneal involvement. The CT-guided biopsies were performed with 1.0-1.8-mm Surecut core-needles. RESULTS: In 16 patients, the final diagnosis was metastatic adenocarcinoma--with the primary tumor sites in the ovary (n = 3), stomach (n = 1), appendix (n = 2), and unknown (n = 10). In the remaining 9 patients, the final diagnosis was hepatocellular carcinoma, lymphoma, and mesothelioma in 1 patient each; tuberculosis in 5; and actinomycosis in 1. Sufficient histological (n = 16) or cytological (n = 8) material was obtained by CT biopsy in 24/25 (96%) cases; the specimen was insufficient for diagnosis in 1 case. In differentiating benign from malignant disease, CT-guided biopsy showed a sensitivity, specificity and accuracy of respectively 89.5%, 100% and 92%. It gave a specific diagnosis in 78.9% (15/19) of patients with malignant conditions and in 50% (3/6) of patients with benign disorders. There were no biopsy-related complications. CONCLUSION: CT-guided percutaneous core-needle biopsy of the omentum is a safe, useful and highly accurate procedure for diagnosing malignant omental pathology. PMID- 9394653 TI - Morphometric measurement of abdominal organs. Comparison of ultrasound and spiral CT. AB - PURPOSE: To determine interobserver variability in the morphometric measurement of abdominal organs with US and spiral CT, and to compare the results obtained with these two modalities. MATERIAL AND METHODS: US and spiral CT examinations of the abdomen were performed in 25 patients. In each patient, 13 defined distances were measured in the liver, spleen and both kidneys with US and spiral CT by two pairs of radiologists in a blinded manner. The interobserver variations of these measurements were evaluated for the US and CT examinations, and the data of both modalities were compared with one another. RESULTS: The measurement of distances in the abdomen with US and spiral CT is subject to considerable interobserver variability in both modalities. The relative interobserver variations showed marked differences, according to which distance was measured. The average interobserver variations were higher in US than in CT. A direct comparison of US and spiral CT revealed that distances obtained with CT frequently exceeded those obtained with US. CONCLUSION: Morphometric measurements of abdominal organs with US and with spiral CT showed considerable differences. The follow-up examinations should therefore be performed with the same imaging modality as used in the original examination. PMID- 9394654 TI - Diagnosis of liver metastases from colorectal adenocarcinoma. Comparison of spiral-CTAP combined with intravenous contrast-enhanced spiral-CT and SPIO enhanced MR combined with plain MR imaging. AB - PURPOSE: The purpose of this study was to determine whether MR with and without SPIO (AMI-25) could replace spiral-CTAP in the staging of colorectal adenocarcinoma. MATERIAL AND METHODS: Thirty-five patients were studied prospectively by means of i.v. contrast-enhanced spiral-CT, spiral-CTAP, and MR of the liver. MR imaging was performed before and after infusion of AMI-25. Diagnoses were compared to intraoperative findings (n = 35) which included intraoperative ultrasound (n = 21), and follow-up CT (n = 18). RESULTS AND CONCLUSION: Fifteen patients were found to have a total number of 53 liver metastases and 43 benign lesions were detected. Evaluation was performed in four different ways: 1) i.v. contrast-enhanced spiral-CT; 2) i.v. contrast-enhanced spiral-CT + spiral-CTAP; 3) plain MR; 4) plain MR + SPIO-enhanced MR. I.v. contrast-enhanced spiral-CT, spiral-CTAP and SPIO-enhanced MR identified patients with liver metastases with equal sensitivity. However, owing to its significantly higher sensitivity, based on a lesion-by-lesion analysis, spiral-CTAP cannot be replaced by SPIO-enhanced MR in patients who are to undergo liver resection. A limitation in spiral-CTAP is its relatively low specificity. PMID- 9394655 TI - Single-session alcohol sclerotherapy in benign symptomatic hepatic cysts. AB - PURPOSE: To evaluate the results of single-session alcohol sclerotherapy in benign symptomatic liver cysts. MATERIAL AND METHODS: Ten cysts (volume 200-4,800 ml) in 10 patients were treated by percutaneous catheterization and injection of 96% ethanol at a dose of 10% of the cyst volume but never more than 100 ml. The treatment was applied for a maximum of 20 min, after which the alcohol and catheter were removed. RESULTS: A satisfactory reduction in cyst volume was achieved in all patients. In 8 patients there was a re-accumulation of fluid during the first period after therapy, followed by a significant reduction in volume on later follow-up examinations. Except for pain, there were no complications. CONCLUSION: Sclerotherapy as a single-session procedure resulted in a significant reduction in cyst volume in all 10 patients. The postprocedural re-accumulation of fluid seen in 8 patients proved to be temporary. It was not necessary to repeat the sclerotherapy procedure in any patient. PMID- 9394656 TI - Human hepatic carbohydrate metabolism. Dynamic observation using 13C MRS without proton decoupling. AB - PURPOSE: Dynamic natural-abundance 13C MR spectroscopy (MRS) studies without proton decoupling were performed in the human liver using commercial 1.5 T MR equipment. MATERIAL AND METHODS: A single tuned custom-made circular surface coil with an OD of 20 cm operating at 16.04 MHz was used for the 13C study. Seventy five grams of glucose dissolved in water was administered for the natural abundance 13C-MRS dynamic study which lasted for approximately 40 to 60 min. Data acquisition was broken into 20-min and 1.7-min blocks. Localized proton shimming with a whole-body coil was performed with sufficient volume to include the observing area of the surface coil; the line width of the water signal was less than 20 Hz. RESULTS AND CONCLUSION: The glucose and glycogen spectra were clearly visible at 80 to 120 ppm after oral administration of the glucose solution. These data demonstrate that dynamic hepatic carbohydrate metabolism can be observed with commercially available MR equipment. Given that the human hepatic glycogen pool reaches maximum level within less than 10 min, this technique should provide a direct diagnosis of hepatic carbohydrate metabolic disorders. PMID- 9394657 TI - CT with different doses of the hepatocyte-specific contrast medium FP 736-03. Evaluation in a nude-rat model of experimental metastases. AB - PURPOSE: To study the dose-response relationship in FP 736-03 (48 mg I/ml), hepatocyte-specific contrast medium for CT of the liver. MATERIAL AND METHODS: A nude-rat model of experimental hepatic metastases was used. CT of the liver was performed before and after i.v. injection of FP 736-03 at 4 different doses: 0.25, 0.5, 1.0 and 2.0 ml/kg b.w. Attenuation in the normal liver parenchyma and in the metastases was measured and plotted as a function of time. RESULTS: The enhancement of the normal liver parenchyma increased in the dose range studied. No increase was found in the metastases: the attenuation here remained constant during the observation period. The maximum enhancement values at doses of 0.25, 0.5, 1.0 and 2.0 ml/kg b.w. were (mean +/- SD) 13.5 +/- 2.7, 30.1 +/- 4.2, 33.2 +/- 4.5 and 59.7 +/- 13.1 HU respectively. PMID- 9394658 TI - Choledochoduodenal fistula secondary to duodenal peptic ulcer. A case report. AB - Spontaneous choledochoduodenal fistula (CDDF) is a rare form of biliary enteric fistula which usually occurs as a complication of duodenal peptic ulcer disease. The more common form is cholecystoduodenal fistula (CCDF) which is generally associated with gallbladder disease. We report on a case of ulcerogenic CDDF diagnosed by upper gastrointestinal barium study, ultrasonography, and gastroduodenal endoscopy. PMID- 9394659 TI - Absorbed dose and image quality in examinations of the colon with digital and analogue techniques. AB - PURPOSE: Image quality and the absorbed dose to the patient are issues of primary interest in the change-over from the conventional analogue technique to the digital technique in the examination of the colon by means of fluoroscopy. The aim of this study was to compare the incident radiation and to evaluate the image quality in two different X-ray equipment types, one digital and one analogue. MATERIAL AND METHODS: A kerma-area product meter was used to measure the incident radiation to the patient. Both fluoroscopy and total-examination times were measured as was the number of images. An evaluation of image quality was made and statistically analysed. RESULTS AND CONCLUSION: No significant difference in the irradiation dose was observed between the two techniques. The fluoroscopy time was significantly lower with the conventional technique but the total-examination time decreased by 18% with the digital technique. The total number of images taken was higher with the digital technique (25 images compared to 19) owing to the limited field of the image intensifier. Significantly more noise and less sharpness were observed with the digital system but there was no significant difference in contrast or image quality in the various anatomical structures. Although the change-over to the digital system produced a reduction in sharpness and an increase in noise, and no significant dose saving was measured, the digital system was faster to work with and could well be used for diagnostic purposes. PMID- 9394660 TI - Unusual imaging presentations in renal transitional cell carcinoma. AB - PURPOSE: To report on unusual imaging presentations in renal transitional cell carcinoma (TCC). MATERIAL AND METHODS: Imaging studies of 140 cases of pathologically proven renal TCC were retrospectively studied with the focus on unusual presentations. RESULTS: Unusual imaging manifestations were found in 20 cases (14.3%). These findings were classified into 5 categories: perirenal abscesses or perirenal hematomas in 6 cases; parenchymal masses in 5; undue thickening of the hydronephrotic wall in 4; "tuberculoid" pyelograms in 3; and tumors with massive necrosis in 2. CONCLUSION: Deceptive imaging presentations may occur in renal TCC. Recognition of these presentations may help to prevent delay in diagnosis. PMID- 9394661 TI - Recurrent varicocele. Demonstration by 3D phase-contrast MR angiography. AB - This is a report on an initial experience with Gd-enhanced 3D phase-contrast MR angiography in 4 patients with recurrent varicocele. The examinations were performed with the patients prone, and a coronal imaging volume was used. The scrotal part of the varicocele was demonstrated in 3 cases and the spermatic vein in 2 cases. This technique could be an alternative to spermatic venography in the radiological mapping of dilated spermatic veins. PMID- 9394662 TI - Pelvic pain syndrome caused by ovarian varices. Treatment by transcatheter embolization. AB - PURPOSE: The aim of this study was to evaluate the clinical effect of therapeutic embolization in the pelvic congestion syndrome caused by ovarian varices. MATERIAL AND METHODS: Six women, aged 25-40 years, with pelvic pain syndrome and marked left (n = 5) or bilateral (n = 1) ovarian varicocele were treated by transcatheter retrograde venous embolization. RESULTS: The pelvic pain syndrome disappeared in all patients within 4 weeks, and there was regression of the periodic pain in 2 women with dysmenorrhoea. The patients were free of symptoms during the 1-4-year follow-up. CONCLUSION: Marked ovarian varices may cause a pelvic pain syndrome. Percutaneous embolization improves both the chronic pain and the dysmenorrhea in these patients. Transcatheter treatment could be considered as an alternative to surgical or laparoscopic ligation in ovarian varicocele. PMID- 9394663 TI - Resistive index of the intrascrotal artery in scrotal inflammatory disease. AB - PURPOSE: To investigate the utility of the resistive indices (RIs) of the epididymal and intratesticular arteries, and to establish diagnostic criteria for scrotal inflammatory disease on the basis of quantitative color Doppler sonography. MATERIAL AND METHODS: We prospectively examined 29 consecutive patients with scrotal pain, and 15 normal control subjects. The RIs of the intratesticular and epididymal arteries were obtained from color Doppler sonographs. RESULTS: The RIs of the testicular artery in epididymoorchitis were significantly lower than those in normal control subjects and in epididymitis (p < 0.01) while the RIs of the testicular artery in epididymitis and control subjects were similar (p > 0.5). With a cut-off value of RI = 0.5, sensitivity, specificity, accuracy, and positive and negative predictive values were 91%, 94%, 94%, 83%, and 77% respectively. The mean RI of the epididymal arteries in epididymitis and epididymoorchitis was 0.49 +/- 0.11. A high level of diagnostic accuracy in scrotal inflammatory disease was achieved when the RIs of the intratesticular and epididymal arteries were less than 0.5 and 0.7 respectively. CONCLUSION: The RI of the intrascrotal artery would give a more objective evaluation than subjective assessment and could provide diagnostic criteria for scrotal inflammatory disease. PMID- 9394664 TI - A case of hyperoxaluria. Radiological aspects. AB - PURPOSE: Oxalosis is an unusual pathological condition with calcium oxalate deposits in soft tissue and bone, recognized as osteosclerosis on radiography. Osteosclerotic bone changes in patients treated with hemodialysis are in most cases due to secondary hyperparathyroidism, but several other diagnoses have to be considered. MATERIAL, METHODS AND RESULTS: We describe the case of a young woman with advanced renal failure treated with hemodialysis since her youth. She had skeletal pain and radiological examination showed: osteosclerosis with sclerotic vertebral bodies; irregular sclerosis and unsharp periostal outline in the tubular bones of the extremities; and acrolysis and calcifications of vascular and soft tissue in the hands. Histological examination showed changes typical of oxalosis. A liver biopsy excluded primary oxalosis type I, and she probably had a secondary oxalosis due to renal failure. This condition (as opposed to primary oxalosis) can be treated with renal transplantation. CONCLUSION: Oxalosis is a rare condition but it should be considered in patients with radiological skeletal changes and chronic renal failure and should not be misinterpreted as renal osteodystrophy. The classification of oxalosis as primary or secondary is important for further treatment. PMID- 9394665 TI - Evaluation of the post-operative lumbar spine with MR imaging. The role of contrast enhancement and thickening in nerve roots. AB - PURPOSE: Two new signs of lumbar nerve-root affection have been reported in recent years on the basis of MR examinations, namely: thickening in nerve roots; and contrast enhancement in nerve roots. The aim of this study was to assess contrast enhancement in nerve roots in a standardised way, and to evaluate the clinical significance of contrast enhancement and of nerve-root thickening in the symptomatic post-operative lumbar spine. MATERIAL AND METHODS: A total of 121 patients (who had previously been operated on for lumbar disc herniation) underwent 152 MR examinations, mainly on a 1.5 T system. Focal nerve-root enhancement was identified by visual assessment. Intradural enhancement was also quantified by pixel measurements that compared the affected nerve roots before and after contrast administration. Non-affected nerve roots were used as reference. RESULTS: Enhanced nerve roots in the dural sac increased at least 40 50% in signal intensity after contrast administration compared to pre-contrast images and also compared to non-affected nerve roots. Intradural nerve-root enhancement was seen in 10% of the patients and focal enhancement in the root sleeve was seen in a further 26%. Nerve-root thickening was seen in 30%. Good correlation with clinical symptoms was found in 59% of the patients with intradural enhancement, in 84% with focal enhancement, and in 86% with nerve-root thickening. The combination of thickening and enhancement in the nerve root correlated with symptoms in 86% of the patients. CONCLUSION: Nerve-root enhancement (whether focal or intradural) and thickening in the nerve root are significant MR findings in the post-operative lumbar spine. In combination with disc herniation or nerve-root displacement, these two signs may strengthen the indication for repeat surgery. However, root enhancement within 6 months of previous surgery may be a normal post-operative finding. PMID- 9394666 TI - Cine-MR imaging of the shoulder. AB - PURPOSE: Shoulder lesions are usually examined with the joint in only one or two positions. We examined the shoulder with the joint in a variety of positions. We also assessed the application of cine-MR to the detection of instability and impingement. MATERIAL AND METHODS: The cine-MR examinations were performed in 30 patients and 15 healthy volunteers. We used an open 0.2 T system and a closed 1.0 T system. Spoiled gradient echo 2D T1-weighted images and turbo spin-echo T1- and T2-weighted images were obtained with a field of view of 180 mm. The examinations were videotaped and evaluated later. RESULTS: Normal variations of the glenohumeral joint were easy to recognize. Sub-luxations and luxations of the humeral head as well as rupture of the labrum were identified. It was also possible to identify the labrum with a signal change after arthroscopic refixation. And we were able to objectively assess distances between the osseous structures during dynamic movement. CONCLUSION: Unlike static MR, cine-MR would appear to be useful in visualizing the capsular ligament complex of the gleno humeral joint in impingement and instability. It also provides information on dynamic changes and may thus prove to be an important tool for shoulder diagnostics. The method may provide an early diagnosis in the sub-acromial impingement syndrome. PMID- 9394667 TI - MR-guided joint puncture and real-time MR-assisted contrast media application. AB - OBJECTIVE: To develop, in MR arthrography of the shoulder joint, an MR-guided technique for localizing the needle puncture and confirming the intracapsular needle-tip position by visualization of the contrast media inflow. MATERIAL AND METHODS: Three unfixed human shoulder specimens were examined on a 1.0 T MR unit. On the basis of MR-compatible markers, the optimal entrance point for puncturing the joint was determined. The precise localization of the needle tip (MR compatible 0.7-mm needle) in the shoulder joint was determined with rapid localizer GRE sequences in 2 orthogonal planes. To confirm the intracapsular position of the needle tip, diluted Gd-DTPA was applied via a long connecting tube and contrast medium inflow into the joint space was controlled on an LCD screen in real-time MR imaging (local-look technique). RESULTS: MR-compatible markers on the skin allowed the rapid determination of the optimal entrance point for needle puncture. An adequate localization of the intra-articular needle-tip position was possible in all specimens although significant artifacts were present on rapid localizer GRE sequences which resulted in an increase in the apparent width of the needle shaft. Real-time MR imaging of the contrast medium inflow was made possible by the local-look technique and LCD screen on the MR unit and this allowed confirmation of the intracapsular position. CONCLUSION: In MR arthrography of the shoulder, an MR-guided technique in conjunction with the LCD screen and real-time MR imaging would seem to be a practical alternative to conventional fluoroscopic guidance. PMID- 9394668 TI - MR imaging of the wrist in carpal tunnel syndrome. AB - PURPOSE: To determine whether specific parameters measured on MR images correlated to electrophysiological changes in carpal tunnel syndrome (CTS). MATERIAL AND METHODS: Prospective clinical examinations were made of 20 patients with suspected CTS. We performed bilateral electrophysiological examinations of the median nerve and bilateral MR imaging of the wrists. RESULTS: The electrophysiological examination suggested median nerve entrapment in 18 wrists. These wrists were compared to the remaining 22 electrophysiologically normal wrists. In addition, we compared both wrists in 12 patients with unilateral symptoms of CTS without reference to the electrophysiological findings. We found no difference in specific MR parameters between the 2 groups. CONCLUSION: Neither symptoms nor electrophysiological findings in CTS were related to specific MR parameters. PMID- 9394669 TI - A simple device for the stereoscopic display of 3D CT images. AB - We describe a simple device for creating true 3D views of image pairs obtained at 3D CT reconstruction. The device presents the images in a slightly different angle of view for the left and the right eyes. This true 3D viewing technique was applied experimentally in the evaluation of complex acetabular fractures. Experiments were also made to determine the optimal angle between the images for each eye. The angle varied between 1 degree and 7 degrees for different observers and also depended on the display field of view used. PMID- 9394670 TI - A new radiographic method for evaluating the degree of sliding in devices used in hip-fracture surgery. AB - PURPOSE: To find a practical method for estimating the degree of sliding that occurs in screw-plate devices used in hip-fracture surgery. Greater understanding of the sliding mechanisms in different fracture types should improve surgical technique and reduce the failure rate. METHODS AND RESULTS: In dynamic screw plate devices, the lag screw slides inside the barrel of the plate. A recent innovation allows the barrel-plate to slide inside a side-plate, thus making possible a combined fracture compression along the neck and the shaft of femur. The lengths of the different parts and the angle of a device in vivo, measured on a radiograph, depend on the position of the femur relative to the photographic film and the roentgen source. We obtained these measurements with a ruler and a protractor from sequential a.p. radiographs of the hip and implemented them in a special computerized program that used the principles of the scaled orthographic and the central projection models. These calculations provided the correct amount of sliding by the lag screw and by the barrel-plate within the side-plate. CONCLUSION: The method presented here can establish the real degree of sliding in screw-plate devices from standard a.p. radiographs independently of the position of the hip. PMID- 9394671 TI - Postero-anterior radiogram of the knee in weight-bearing and semiflexion. Comparison with MR imaging. AB - PURPOSE: The purpose was four-fold: to assess the reproducibility of p.a. weight bearing radiograms of the knee and the minimal joint-space (MJS) width measurements in these radiograms; to compare the MJS with MR-detected cartilage defects; to evaluate the location of these cartilage defects; and to estimate the relation between meniscal abnormalities and joint-space narrowing. MATERIAL AND METHODS: Fifty-nine individuals, aged 41-58 years (mean 50), with chronic knee pain were examined by means of p.a. weight-bearing radiograms in semiflexion with fluoroscopic guidance of the knee joint. The MJS was measured with a standard ruler. On the same day MR imaging was performed with proton-density- and T2 weighted turbo spin-echo on a 1.0 T imager. Meniscal abnormalities and cartilage defects in the tibiofemoral joint (TFJ) were noted. RESULTS AND CONCLUSION: The p.a. view of the knee and the MJS measurements were reproducible. MJS of 3 mm is a limit in diagnosing joint-space narrowing in knees with MR-detected cartilage defects. There was a high proportion (p < 0.001) of meniscal abnormality within the narrowed compartments in comparison with those that were not narrowed. A larger number of the cartilage defects (p < 0.05) was found in the medial femoral condyle than in any of the other condyles of the TFJ. The defects had a dorsal location (p < 0.001) as shown in the weight-bearing radiograms of the knee in semiflexion. PMID- 9394673 TI - Correlation of T1 and T2 relaxation rates in normal bone-marrow water with serum ferritin concentration. AB - PURPOSE: This study was made to clarify the paramagnetic effect of iron stored in the hematopoietic tissue of bone marrow. MATERIAL AND METHODS: The T1 and T2 relaxation times of bone-marrow water in the L1-3 vertebrae of 20 healthy individuals were measured by MR imaging with a 1.5 T magnet. The chemical shift misregistration effect was used to isolate the bone-marrow water. The results were compared with the serum ferritin concentration. RESULTS AND CONCLUSION: Although no correlation between the T1 relaxation rate of the water fraction and the serum ferritin concentration was evident, the T2 relaxation rate of the water fraction showed strong linear correlation with the serum ferritin concentration (r = 0.87, p < 0.0001). Thus, T2 of bone-marrow water accurately reflects the amount of iron in normal bone marrow. This finding may be useful in the evaluation of the characteristics of hematopoietic tissue in bone marrow. PMID- 9394672 TI - Magnetization transfer and spin lock MR imaging of patellar cartilage degeneration at 0.1 T. AB - PURPOSE: To investigate magnetization transfer (MT) parameters and rotating frame relaxation time T1 rho in patellar cartilage at different levels of degeneration. MATERIAL AND METHODS: Thirty cadaveric patellae were examined at 0.1 T using the time-dependent saturation-transfer MT technique and the spin lock (SL) technique. In an SL experiment, nuclear spins are locked with a radiofrequency (RF) field, and the locked nuclear magnetization relaxes along the magnetic component of the locking RF field. The specimens were divided into three groups according to the level of cartilage degeneration. MT parameters and T1 rho were measured. RESULTS: The MT effect was greater in degenerated cartilage than in normal cartilage. T1 rho was longer in advanced cartilage degeneration than in intermediate cartilage degeneration. CONCLUSION: The results suggest that more studies are needed to fully establish the value of SL imaging in cartilage degeneration. PMID- 9394674 TI - Urine viscosity after injections of iotrolan or iomeprol. AB - PURPOSE: Previous observations in rats caused us to speculate whether the injection of iotrolan, a nonionic dimeric contrast medium (CM), would increase urine viscosity enough to obstruct urine outflow in the collecting duct. MATERIAL AND METHODS: The urine viscosity in dogs was measured directly with a viscosimeter after injections of iotrolan or of iomeprol, a nonionic monomeric CM. RESULTS AND CONCLUSION: The injection of iotrolan increased urine viscosity considerably whereas iomeprol had little effect on this variable. The osmolality dependent adverse reactions of CM have previously been emphasized but viscosity dependent adverse reactions must also be considered when the CM is a polymer with a low osmolality. PMID- 9394675 TI - An in vitro 1H-MRS model of oncogene transfection. The spectral feature of c-erbB 2 and c-Ha-ras transfected NIH3T3 fibroblast cells. AB - PURPOSE: Malignancy is an abnormality of cell division and differentiation based on abnormal expression of oncogenes. This note describes the in vitro 1H-NMR spectral features of oncogene-transfected NIH3T3 fibroblast cells compared to non transfected cells. MATERIAL AND METHODS: 1H-NMR spectra of cultured NIH3T3 cells and c-erbB-2 or c-Ha-ras gene-transfected cells were obtained by 400 MHz high resolution NMR. The peaks were assigned by 2D HOHAHA spectra of the cell suspension and the spectral changes were evaluated in 1D and 1D differential spectra. RESULTS: The 1H spectra obtained from both transfected cell lines were broadened over all peaks, suggesting reduced mobility in plasma membrane lipid molecules. No other differential spectra for characterizing metabolic change was detected. CONCLUSION: Broadened 1H spectra observed after c-erbB-2 or c-Ha-ras transfection suggest changes of plasma membrane viscosity, which may be related to the oncogene expression. PMID- 9394676 TI - Combinations of nonlabeled, 125I-labeled, and anti-idiotypic antiplacental alkaline phosphatase monoclonal antibodies at experimental radioimmunotargeting. AB - PURPOSE: Placental alkaline phosphatase (PLAP) is a membrane-bound oncofetal antigen that can be used for radioimmunotargeting. Preinjection of nonlabeled monoclonal anti-PLAP antibody (H7) and postinjection of monoclonal anti-idiotypic anti-PLAP antibody (alpha H7) were used in order to improve the localization efficacy of 125I-labeled H7. MATERIAL AND METHODS: A human cervix adenocarcinoma cell line (HcLa Hep 2) was inoculated subcutaneously in 24 nude mice. Repeated quantitative radioimmunoscintigraphic recordings were performed on 27 occasions in each of the 24 mice during the observation period which lasted for nearly 3 months. The tumor and nontumor doses were calculated according to the Medical International Radiation Dose Committee formula on the basis of the scintigraphic data. RESULTS: All tumors were clearly visualized as early as one day after injection of 125I-labeled H7. The remaining radioactivity was exclusively located in the tumors at days 30-81. As much as 12-16% of the injected dose/g accumulated in the tumors during the first 2 days after injection, and remained stable at this high level for approximately 10 days in all investigated groups. Radioactivity in the whole body was rapidly eliminated during the same time period. The highest tumor/nontumor dose ratio was obtained after a single injection of 125I-labeled H7. CONCLUSION: Neither a preinjection of nonlabeled H7 nor a postinjection of alpha H7 nor a combination of both strategies resulted in improved tumor/nontumor dose ratios compared to a single injection of labeled H7. The monoclonal antibody H7 has a rapid and high uptake, combined with a prolonged retention time in the tumors. The kinetic properties of H7 are different from antibodies targeting intracellular tumor antigens. PMID- 9394677 TI - Bioavailability of folic acid in fortified food. PMID- 9394678 TI - Retinoids and alcoholic liver disease. PMID- 9394679 TI - Complementary medicine: a review of immunomodulatory effects of Chinese herbal medicines. AB - Popular demand for and scientific interest in complementary or alternative medicine, particularly medicinal botanicals, has increased considerably in recent years. The medicinal botanicals with the longest tradition, and for which extensive data are available, are Chinese herbal medicines and their Japanese counterparts--Kampo medicines. This review focuses on some representative examples of studies examining the effects of some traditional Chinese medicines on various aspects of the immune response. In vitro as well as in vivo studies are cited, the latter including not only animal experiments but also clinical trials. Although by no means exhaustive, this review attempts to show that much research has focused on the specific beneficial effects of Chinese herbal medicines. Studies examining the mechanisms by which they exert their immunomodulatory actions, however, are found much less frequently. Nonetheless, even the limited number of mechanistic experiments presented here reveal that numerous mechanisms are likely involved in the various actions of even a single medicine. It will be the elucidation of such mechanisms that will provide the scientific basis for establishing the efficacy and safety of not only Chinese herbal medicines but all forms of medicinal botanicals. PMID- 9394680 TI - New definitions in nutritional disciplines: will the public be damned or saved? PMID- 9394681 TI - Changes in total body fat during the human reproductive cycle as assessed by magnetic resonance imaging, body water dilution, and skinfold thickness: a comparison of methods. AB - Total body fat and fat-free mass were assessed by magnetic resonance imaging (TBFMRI and FFMMRI) in 11-16 healthy Swedish women before pregnancy and 5-10 d and 2, 6, and 12 mo after delivery. On these occasions, TBF was also measured by the body water dilution (TBFBWD) and skinfold-thickness (TBFSFT) techniques. The results were used to compare changes in TBFSFT and TBFBWD during reproduction with changes in TBFMRI. TBFBWD was 1.5-4.0 kg higher than TBFMRI and at all postpartum measurements the difference between these estimates increased significantly with increased body fat content. This difference was also significantly higher 6 mo after delivery than it was 2 and 12 mo postpartum. The possibility that this was due to variations in the degree of hydration of FFM postpartum was considered. TBFSFT was 1.7-3.1 kg higher than TBFMRI and this difference increased with increasing body fat content. The agreement between changes in TBFMRI and TBFSFT was different during different times in the reproductive cycle and was also influenced by the amount of fat lost or gained. The findings thus suggest that there is a risk for bias when changes in TBF during reproduction are estimated by the skinfold-thickness technique as well as by isotope dilution. PMID- 9394682 TI - Body composition of a young, multiethnic, male population. AB - The study objective was to establish the range of total body-composition values for a young, multiethnic, healthy male population (aged 3-18 y) by using dual energy X-ray absorptiometry (DXA). Results for 297 males in three ethnic groups [European American (white), n = 145; African American (black), n = 78; and Mexican American (Hispanic), n = 74] are reported. Bone mineral content (BMC), lean tissue mass (LTM), body fat mass, and percentage fat are presented as functions of age. Analysis of variance with age, weight, and height as covariates was used to evaluate differences among the three ethnic groups. BMC and LTM were higher in black than in white males, but no difference in BMC or LTM was evident between the white and Hispanic groups. The relation between total-body BMC and LTM was linear (r = 0.985, P < 0.0001) and independent of age or ethnic classification. The Hispanic males had higher body fat values than the white group, whereas the black males generally had lower values than the white group. When adjusted for body size, the Hispanic males continued to have significantly higher body fat and percentage fat than the white or black males. Ethnic-specific equations for the prediction of body composition as a function of age, weight, and height were derived. The results for the white males in the present study were compared with DXA-derived reference data reported in other countries for young white males. We conclude that reference values of total body composition for young healthy males need to be ethnic specific. PMID- 9394683 TI - Effect of high-fat and low-fat diets on voluntary energy intake and substrate oxidation: studies in identical twins consuming diets matched for energy density, fiber, and palatability. AB - There remains controversy over the effects of dietary fat content on voluntary energy intake. Additionally, the question of whether there is a genetic susceptibility to overeating high-fat diets has not been resolved. To address these issues, we designed two diets: a low-fat diet providing approximately 20% of energy as fat and a high-fat diet with approximately 40% of energy as fat. The diets were matched for energy density, fiber, and palatability. In a two-phase, 18-d intervention study, voluntary energy intakes and macronutrient oxidation rates during the fasting and fed states were determined in seven pairs of identical male twins. In contrast with results of previous intervention studies, in which low-fat and high-fat diets were not matched for energy density and other associated variables, we observed no significant difference in voluntary energy intake between the low-fat and high-fat phases, and mean daily intakes were similar (10.3 and 10.7 MJ/d, respectively). Postprandial rates of fat oxidation tended to reflect fat intakes in the two dietary phases, thus helping to explain the lack of a difference in mean energy intakes. There was also a significant twin-pair similarity in differences in energy intakes between dietary phases (P = 0.013). These results suggest that dietary fat content does not have a major influence on voluntary energy intake when dietary variables usually associated with fat are controlled for and that there may be a familial influence on the effects of dietary fat content on energy intake. PMID- 9394684 TI - Leptin concentration in women is influenced by regional distribution of adipose tissue. AB - Leptin concentrations in humans are increased with obesity, and women have higher leptin concentrations than men. This sex difference reflects the greater fat mass of women. However, there is evidence that factors other than the size of the adipose tissue mass contribute to serum leptin concentrations. This study was undertaken to determine whether anthropometric factors influenced leptin concentrations in our population. Leptin concentrations were measured in 375 persons from a population study of hypertension and diabetes for whom body composition data (bio-electrical impedance analysis and anthropometry) were available. Serum leptin concentrations were more than four times higher in women than in men (18.5 +/- 13.9 compared with 3.8 +/- 3.6 ng/L, P < 0.0001). In individuals with comparable body mass indexes, these differences persisted after adjustment for either percentage fat (P < 0.05) or fat mass (P < 0.0001) by multivariate-regression analysis. After fat mass was adjusted for, the serum leptin concentration in both men and women was independent of waist circumference but in women was associated with hip circumference. Hip circumference is a proxy measure of peripheral fat and these results suggest that the larger hips of women may contribute to the sex difference in serum leptin concentration. PMID- 9394685 TI - Intraabdominal adipose tissue and anthropometric surrogates in African American women with upper- and lower-body obesity. AB - It is well established that visceral adipose tissue (VAT) is associated with increased morbidity and mortality in white women. In a recent study, we found that African American women had smaller depots of VAT. To test the relation of VAT to the commonly used anthropometric surrogates for fat patterning, including waist-to-hip ratio (WHR), waist circumference, subscapular skinfold thickness, and ratio of triceps to subscapular skinfold thickness, we recruited 48 normotensive African American women > 120% of ideal body weight on the basis of WHRs > 0.85 [upper-body obesity (UBO); n = 23] and < 0.76 [lower-body obesity (LBO); n = 25]. There were no differences between groups in age, height, weight, body mass index, or percentage of body fat. VAT was determined by magnetic resonance imaging at L4-5; percentage of fat was determined by dual-energy X-ray absorptiometry. Women with UBO had significantly larger mean (+/- SEM) depots of VAT at L4-5 than did women with LBO (0.26 +/- 0.02 compared with 0.19 +/- 0.02 L). Waist circumference was the single best predictor of VAT at L4-5 in both groups of women whereas WHR was significantly associated with VAT at L4-5 only in women with UBO. In African American women, waist circumference is a better surrogate for VAT than is WHR. PMID- 9394686 TI - Effects of fasting and glucose infusion on basal and overnight leptin concentrations in normal-weight women. AB - The plasma concentration of leptin is reduced in association with chronic energy restriction and weight loss in humans, but little is known about the acute effects of fasting and glucose infusion on leptin. In this study, plasma leptin, insulin, glucose, and fatty acid concentrations were measured daily in 14 healthy, normal-weight, female volunteers aged 24 +/- 4 y with a body mass index (kg/m2) of 24.2 +/- 3.6 during a 4-d fast. The mean plasma leptin concentration decreased by 54 +/- 8% with fasting (P = 0.0006, ANOVA). In a stepwise-regression model, the change in leptin concentration with fasting correlated most significantly with the change in insulin (R2 = 0.48, P = 0.0057) and to a lesser extent with the change in body fat by bioimpedance analysis (R2 = 0.19, P = 0.03). Plasma leptin concentrations measured every 20 min from 2000 to 0800 on the fourth night of the fast did not show a time-dependent rise. A continuous intravenous infusion of 5% glucose providing 1414 +/- 323 kJ/d (338 +/- 78 kcal/d) was begun after 4 d of fasting in seven subjects who continued to fast for an additional 6 d. Within 24 h of the glucose infusion, leptin concentrations increased significantly by 80 +/- 52% (P < 0.05). These data show the sensitivity of plasma leptin concentrations to small changes in energy supply and suggest a basic role of substrate metabolism in the short-term regulation of leptin. PMID- 9394687 TI - Three-month nutritional supplementation in Indonesian infants and toddlers benefits memory function 8 y later. AB - Does short-term supplementary feeding during infancy and childhood have long lasting effects? In 1986, 334 children aged 6-60 mo living on rural tea plantations in West Java, Indonesia, participated in a 3-mo randomized trial to test the effects of a dietary supplement providing approximately 1672 kJ (400 kcal) energy/d, with about the same nutrient density as local foods. We returned to the same communities in 1994 and enrolled 231 (125 supplemented, 106 control) of the original subjects in a follow-up study of the long-term effects of supplementation. We assessed these subjects by using several measures: anthropometry, iron status, information processing, Peabody Picture Vocabulary Test, word fluency, and an arithmetic test. The supplemented group showed no differences from those in the control group. However, when the analysis was limited to subjects who had received the supplement before the age of 18 mo (n = 73), the supplemented children performed better than control children on the Sternberg test of working memory (decision time intercept: probe absent, P = 0.002; probe present, P = 0.053). After considering possible confounders, we concluded that the supplementation during infancy was responsible for the difference. This finding shows that supplementation can have long-lasting effects on a specific domain if the child receives it at the appropriate stage of development. PMID- 9394688 TI - Clinical outcome of geriatric patients in the United States receiving home parenteral and enteral nutrition. AB - In this study the use of home parenteral and enteral nutrition (HPEN) therapy in geriatric patients and the effect of aging on the clinical outcome of HPEN therapy was assessed between 1985 and 1992. Data were obtained from Medicare (part B) parenteral and enteral nutrition workload statistics, Blue Cross and Blue Shield of South Carolina, and the North American HPEN Patient Registry. On the basis of these data it was estimated that in 1992 there were 40,000 HPN patients and 152,000 HEN patients nationwide. One-quarter to one-third of the HPN group was aged > or = 65 y, depending on the underlying diagnosis. Geriatric HPN patients had a generally good outcome but did not do as well as their younger counterparts; however, they had fewer therapy-related complications than children. In the HEN group, 44% of cancer patients and 69% of patients with neuromuscular swallowing disorders were geriatric. Geriatric HEN patients with swallowing disorders had a poorer outcome (i.e., lower survival, poorer rehabilitation, and fewer resumed full oral nutrition) than younger patients after 1 y of follow-up. In conclusion, although aging was associated with a poorer outcome for HPN patients, it was still reasonably good; therefore, age per se should not exclude geriatric subjects from therapy. For HEN, the poorer outcome in geriatric dysphagic patients raises concern regarding the appropriateness of this therapy. PMID- 9394689 TI - Maternal obesity and breast-feeding success in a rural population of white women. AB - Maternal obesity interferes with the initiation and maintenance of lactation in animal models but it has not been investigated widely in women. We reviewed medical records from a white population to examine the relation between prepregnant overweight [body mass index (BMI; in kg/m2) 26.1-29.0] and obesity (BMI > 29.0) on initiation and duration of breast-feeding. Logistic regression revealed that of those who ever put their infants to the breast (n = 810), women who were overweight [odds ratio (OR) = 2.54, P < 0.05] or obese (OR = 3.65, P < 0.0008) had less success initiating breast-feeding than did their normal-weight counterparts (BMI < 26.1). Proportional-hazards regression revealed higher rates of discontinuation of exclusive breast-feeding in overweight (RR = 1.42, P < 0.04) and obese (RR = 1.43, P < 0.02) women and higher discontinuation of breast feeding to any extent in overweight (RR = 1.68, P < 0.006) and obese (RR = 1.73, P = 0.001) women. Controlling for parity, socioeconomic status, maternal education, and other factors that often covary with maternal obesity and breast feeding did not change these results. These results suggest that excessive fatness in the reproductive period may inhibit lactational performance in women. PMID- 9394690 TI - Changes in vitamin B-6 status indicators of women fed a constant protein diet with varying levels of vitamin B-6. AB - Changes in vitamin B-6 status indicators were evaluated in vitamin B-6-replete subjects. Ten young women consumed diets providing 85 g protein/d and 1.03, 1.33, 1.73, and 2.39 mg vitamin B-6/d for 12 or 15 d during four successive diet periods; in a second study, six women were fed diets providing 85 g protein/d and 0.84, 1.14, and 2.34 mg vitamin B-6/d for 10 or 12 d during three successive diet periods. Vitamin B-6 status indicators showing significant differences among intakes included urinary excretion of 4-pyridoxic acid and total vitamin B-6, pyridoxal 5'-phosphate and total vitamin B-6 in plasma, and xanthurenic acid excretion after a 2-g L-tryptophan load. Significant correlations were found between vitamin B-6 intake and 4-pyridoxic acid, total vitamin B-6, plasma pyridoxal 5'-phosphate, plasma total vitamin B-6, erythrocyte alanine aminotransferase percentage stimulation and postload excretion of xanthurenic acid and volatile amines (kynurenine plus acetylkynurenine). Depending on the indicator, between 20% and 70% of the subjects had inadequate values for 4 pyridoxic acid, total vitamin B-6, plasma pyridoxal 5'-phosphate, and erythrocyte alanine aminotransferase percentage stimulation at a vitamin B-6 intake of 1.33 mg/d (0.016 mg vitamin B-6/g protein). A ratio of dietary vitamin B-6 to protein > 0.016 mg/g is required for adequate vitamin B-6 status in women. PMID- 9394691 TI - Absorption of folate from fortified cereal-grain products and of supplemental folate consumed with or without food determined by using a dual-label stable isotope protocol. AB - The absorption of folic acid in fortified white and whole-wheat bread, rice, or pasta or in solution was evaluated in human subjects with use of a single-dose, dual-label, stable-isotope protocol that did not involve prior loading of subjects with nonlabeled folate. In each of five sequential trials, 14 adults received a single oral dose of [13C5]folic acid in one of the four fortified cereal-grain products or in water concurrently with an intravenous injection of [2H2]folic acid. In two additional trials, subjects received oral [13C5]folic acid with or without a light breakfast meal. In all trials, urine was collected 24-36 h postdosing and the isotopic labeling of urinary folates determined. Isotope excretion ratios of urinary folates (% [13C5]folate dose/% [2H2]folate dose), which were used as criteria of absorption, indicated no significant differences among the various fortified foods and the control (P = 0.607). Because statistical power was sufficient to have detected a 50% difference from the control, these results suggest that [13C5]folic acid in these fortified cereal-grain foods was highly available. This study also suggests that fortification will contribute effectively to the folate status of the population. Consuming [13C5]folic acid after a light breakfast meal led to a small reduction in absorption relative to the control without food (P < 0.085). Between-subject variation in this protocol exceeded that observed in previous studies conducted using prior saturation of subjects with nonlabeled folic acid. We recommend that either prior saturation or multiple doses be used in future applications of this technique to improve precision. PMID- 9394692 TI - Field-study screening of blood folate concentrations: specimen stability and finger-stick sampling. AB - We describe optimized procedures for field studies of blood folate concentrations by using finger-stick blood sampling and include relevant studies on blood folate stability. We introduce whole-blood folate adjustment using sample hemoglobin (folate/hemoglobin, nmol/g) as a novel and practical tool yielding accurate and precise results when blood volume or dilution is unknown. Red cell folate concentrations (nmol/L) of 11,887 Americans correlated well with hemoglobin corrected whole-blood folate concentrations (r2 = 0.993; red cell folate = 0.347 x hemoglobin folate + 1 nmol/L), which supports the approach of using the mean cell hemoglobin concentration (g/L) to interconvert red cell and hemoglobin folate data. Folate concentrations in capillary (finger stick) and venous blood samples from 28 normal donors were similar (P > 0.87), correlating closely (r = 0.98, P < 0.001). Whole-blood samples (collected into K2-EDTA-containing evacuated tubes) in field studies are best stored intact at 4 degrees C until they can be processed and frozen (-20 degrees C). Specific knowledge of blood folate stability is essential in planning and designing field studies. PMID- 9394693 TI - A 14-mo zinc-supplementation trial in apparently healthy Chilean preschool children. AB - Apparently healthy preschool children (46 boys, 52 girls) aged 27-50 mo from low socioeconomic conditions who attended daycare centers in Santiago participated in a 14-mo long double-blind zinc supplementation trial. Unlike most previous studies, no additional inclusion criteria such as short stature or slow growth rate were considered. Subjects were pair matched according to sex and age and randomly assigned to two experimental groups: the supplemented group, which received 10 mg Zn/d, and the placebo group. Selected anthropometric, clinical, dietary, biochemical, and functional indexes were determined at the beginning of the study and after 6 and 14 mo of intervention. Actual dietary zinc intake was 66% of the recommended dietary allowance. Height gain after 14 mo was on average 0.5 cm higher in the supplemented group (P = 0.10). The response, however, was different between sexes. Boys from the supplemented group gained 0.9 cm more than those in the placebo group (P = 0.045). No effect was seen in girls. Although no significant differences were observed in the rest of the variables studied, trends (0.05 < P < 0.10) in the supplemented group compared with the placebo group for increased midarm muscle area in boys, improved response to tuberculin, and reduced rates of parasite reinfestation were noted. We conclude that in preschool children of low socioeconomic status, zinc is a limiting factor in the expression of growth potential. PMID- 9394694 TI - Association between dietary fiber intake and the folate status of a group of female adolescents. AB - The main objective of this study was to assess the association between dietary fiber intake and the folate status of Canadian female adolescents. We also assessed dietary folate intakes and evaluated the prevalence of biochemical folate deficiency in these subjects. Female adolescents aged 14-19 y (n = 224) were recruited and fasting blood samples were collected. Dietary intakes (3-d food record) were recorded and participants were classified as lactoovovegetarians, semivegetarians, or omnivores on the basis of food consumption patterns assessed with food-frequency questionnaires. Fourteen percent, 17%, and 26% of lactoovovegetarians, semivegetarians, and omnivores, respectively, had dietary folate intakes below their predicted requirements; 1%, 4%, and 23%, respectively, had serum folate concentrations indicative of deficiency. Despite low dietary folate intakes and serum folate concentrations, few subjects had homocysteine concentrations indicative of deficiency, suggesting that the degree of folate depletion had not yet produced functional consequences. Most important, results suggest that the consumption of nonstarch polysaccharide is significantly associated with serum folate concentrations (P < 0.001). For each 1-g increase in nonstarch polysaccharide intake, a 1.8% increase in serum folate concentration is expected. In summary, we propose that an increase in nonstarch polysaccharide intake may promote the intestinal biosynthesis of folate, providing a complementary strategy to enhance the folate nutriture of humans. PMID- 9394695 TI - Effect of intestinal parasite treatment on the efficacy of oral iodized oil for correcting iodine deficiency in schoolchildren. AB - Oral supplementation with iodized oil for correction of iodine deficiency in a population has advantages over intramuscular injection but the duration of effect is shorter. The relation of intestinal parasite treatment and efficacy of oral iodized oil was examined in an intervention study in 8-10-y-old schoolchildren in Malawi. Severely iodine-deficient schoolchildren with a single parasitic infestation of Ascaris lumbricoides (n = 44), hookworm (n = 42), or Entamoeba histolytica (n = 24) were randomly allocated to receive or not receive treatment before taking a 1-mL oral supplement (490 mg I) of iodized ethyl esters from poppy seed oil. The urinary iodine concentration was measured at various time points after supplementation to define the time intervals before urinary iodine concentrations returned to 0.40 mumol/L, indicating moderate iodine deficiency. Treatment with metronidazole for E. histolytica increased the protection period from 2.0 to 21.0 wk (P < 0.05). For all untreated children together, the duration of effect was 9.2 wk shorter (P < 0.001) than that for their treated peers (16.8 wk). We conclude that intestinal parasitic infestations reduce the efficacy of oral supplementation with iodized ethyl esters by interfering with absorption. PMID- 9394696 TI - Sex differences in response to dietary manipulation in rats with hypertension and myocardial hypertrophy. AB - Studies of the effect of sex on the metabolic state of rats with chronic hypertension and concomitant myocardial hypertrophy were conducted. Female and male spontaneously hypertensive rats (SHRs) with early myocardial hypertrophy (5.5 mo old) were used. Serum fatty acids, liver glycogen, and myocardial glycogen were measured at baseline and after the rats were deprived of food for 24 h. The metabolic effects of progressive myocardial hypertrophy in females were assessed in additional groups of female SHRs (5.5 or 12 mo old) under the following conditions: control, food deprived, or food deprived and refed with equienergetic lipid-rich (38.9% of total energy) or carbohydrate-rich (76.5% of total energy) diets. Despite no differences in serum fatty acids, females had significantly higher baseline myocardial glycogen and liver glycogen concentrations than males. In response to food deprivation, females continued to have significantly higher myocardial glycogen and fatty acid concentrations than males, whereas there were no sex differences in liver glycogen, which was depleted in both males and females. Older hypertensive females had higher baseline fatty acid concentrations and lower liver glycogen concentrations than younger females, whereas there were no differences in myocardial glycogen. Food deprivation doubled fatty acid concentrations, depleted liver glycogen, and increased myocardial glycogen in both age groups. In both age groups, fatty acid concentrations and liver glycogen did not return to baseline values after food deprivation and refeeding. In both age groups, fatty acid concentrations increased further after the lipid-rich diet whereas liver glycogen concentrations returned to approximately 50% of baseline values after the carbohydrate-rich diet. Refeeding with either diet did not significantly increase myocardial glycogen further. Thus, the metabolic response to dietary manipulation was influenced by both sex and, in females, progressive pathology. PMID- 9394697 TI - Body composition in adults with cerebral palsy by dual-energy X-ray absorptiometry, bioelectrical impedance analysis, and skinfold anthropometry compared with the 18O isotope-dilution technique. AB - The aim of this study was to determine in adults with cerebral palsy the accuracy and practicality of standard methods used to estimate body composition. The sample consisted of 20 adults (13 men and 7 women) aged 20-55 y with various degrees of cerebral palsy. Percentage body fat was estimated from skinfold thickness, bioelectrical impedance analysis (BIA), and dual-energy X-ray absorptiometry (DXA), and compared with the reference measure of percentage body fat from total body water by 18O dilution. Values derived from use of BIA and skinfold thickness, estimated by using the Jackson-Pollock equation, were significantly different from those derived with use of 18O dilution (P < 0.001 and P < 0.001, respectively). There was no significant difference between percentage body fat measured with DXA and that measured with 18O. There was favorable agreement between DXA and 18O (mean difference: 0.06 +/- 9.6%), but not between skinfold thickness (mean difference: 6.33 +/- 12.3%) or BIA (mean difference: -6.55 +/- 13.6%) and 18O. Although DXA was the best measure for predicting percentage body fat in the sample, its high cost prohibits its use as a practical method. The best-fitting regression equation specific for this sample by using anthropometric measures to predict percentage body fat was as follows: 8.76 - (7.34 x sex) + (0.32 x weight) + (0.38 x biceps skinfold) (R2 = 0.84, P < 0.001, SEE = 4.85). This equation needs to be cross-validated in an independent sample of adults with cerebral palsy. PMID- 9394698 TI - Effects of lowering fat and increasing dietary fiber on fasting and postprandial plasma lipids in hypercholesterolemic subjects consuming a mixed Mediterranean Western diet. AB - The aim of this study was to evaluate the cholesterol-lowering effects of reducing fat and increasing or not increasing dietary fiber in subjects consuming a mixed Mediterranean-Western diet. Thirty-one free-living, mildly hypercholesterolemic subjects were randomly allocated to two groups. Subjects in both groups first shifted for 4 wk to a low-fat, low-fiber diet (LFLFD). For an additional 4-wk period, subjects in group 1 continued consuming the LFLFD whereas subjects in group 2 consumed a low-fat, high-fiber diet (LFHFD). Most dietary fatty acids were monounsaturated (38-41%) and fibers, when provided (up to 35 g/d), came from unrefined cereals, legumes, and soluble-fiber-enriched ready-to eat cereals. After period 1 of the LFLFD, mean serum and low-density-lipoprotein (LDL)-cholesterol concentrations of subjects in groups 1 (-12.5% and -15.5%, respectively) and 2 (-10.5% and -15.5%, respectively) decreased significantly from baseline (P < 0.05). After period 2, mean serum and LDL-cholesterol concentrations of subjects consuming the LFLFD (group 1) were still lower (by 8.8% and 9.2%, respectively, from baseline) whereas in subjects consuming the LFHFD (group 2) these values decreased further to significantly lower values (14.2% and 17.6% from baseline, respectively). Fasting high-density-lipoprotein (HDL) cholesterol, apolipoprotein A-I, glycemia, and insulinemia did not change significantly. In seven men, postprandial lipemia transiently increased more after a breakfast test meal at the completion of the LFHFD period than after the LFLFD period. In conclusion, an LFHFD more comparable with the traditional Mediterranean diet may improve the dietary management of moderate hypercholesterolemia. PMID- 9394699 TI - Hormonal implications of the hypocholesterolemic effect of intake of field beans (Vicia faba L.) by young men with hypercholesterolemia. AB - This study examined the hypocholesterolemic effect and hormonal changes resulting from 30 d of supplementation with Vicia faba L. (field bean) flour of diets of young men (aged 18-21 y; n = 40) with borderline-high or high serum cholesterol values. All subjects (groups A-D) consumed the same basic diet. Additionally, volunteers in the control group (A) consumed 90 g control flour/d whereas those in the three bean groups received either 90 g cooked field bean flour (groups B and C) or 90 g raw field bean flour (group D) daily. Groups A and B included volunteers with borderline-high cholesterol values [5.2-6.2 mmol total cholesterol/L and 3.4-4.1 mmol low-density-lipoprotein (LDL) cholesterol/L]. Subjects in groups C and D had high serum cholesterol concentrations (total cholesterol > 6.2 mmol/L and LDL cholesterol > 4.1 mmol/L). After 30 d, serum glucose, insulin, triacylglycerol, total, LDL-cholesterol, and very-low-density lipoprotein (VLDL)-cholesterol values were significantly lower than initial values in all subjects who consumed diets containing field bean flour (P < or = 0.0001, except for LDL-cholesterol concentrations in group C, for which P < or = 0.0007). Legume intake also resulted in a significant increase (P < or = 0.0001) in glucagon and high-density-lipoprotein cholesterol. Neither cortisol nor thyroid hormone values changed significantly. The results suggest that the hypocholesterolemic effect of field bean intake depends at least partly on a concomitant increase in glucagon and decrease in insulin values. The more marked reduction in triacylglycerol and VLDL-cholesterol concentrations in subjects who consumed raw field beans indicates a coparticipation of their thermolabile components. PMID- 9394700 TI - beta-Carotene beadlets potentiate hepatotoxicity of alcohol. AB - Administration of beta-carotene in beadlets to baboons potentiates alcohol induced liver injury. To determine whether this also occurs in other species, and whether the beadlet carrier itself contributes to the toxicity, rats were given for 2 mo vitamin A (1.4 U/J), beta-carotene (4.8, 12.0, and 24.0 U/J, with or without beadlets), or corresponding amounts of beadlets without beta-carotene, in diets containing either carbohydrates or equivalent amounts of ethanol. Isoenergetic substitution of ethanol (36% of total energy) for carbohydrates reduced hepatic vitamin A by 80%, and such a depletion was also observed with beta-carotene as vitamin A precursor. By contrast, ethanol raised hepatic beta carotene, which was further increased by beadlets. Thus, whereas alcohol promoted hepatic depletion of vitamin A, it had the opposite effect on beta-carotene. Ethanol seems to affect the homeostasis of beta-carotene. Furthermore, the ethanol-induced oxidative stress, assessed by an increase in hepatic 4 hydroxynonenal and F2-isoprostanes (measured by gas chromatography-mass spectrometry), was not improved despite a concomitant rise in hepatic antioxidants (beta-carotene and vitamin E). Moreover, beadlets resulted in proliferation of the smooth endoplasmic reticulum and in leakage of the mitochondrial glutamate dehydrogenase into the plasma, reflecting mitochondrial injury (both documented by electron microscopy). Thus, in rats given ethanol, beta-carotene is not an efficient vitamin A precursor. Its bioavailability was improved by beadlets, but the ethanol-induced oxidative stress was not attenuated and there was associated hepatotoxicity that now needs to be assessed in humans. PMID- 9394701 TI - Low dietary fiber and high protein intakes associated with newly diagnosed diabetes in a remote aboriginal community. AB - The high prevalence of diabetes mellitus in North American aboriginal populations may be due to recent changes in lifestyle, including the adoption of a high-fat, low-fiber diet. To determine whether fat or fiber intakes were associated with new cases of diabetes, we studied 72% (728/1018) of residents aged > 9 y from a remote aboriginal community in northern Ontario using the 75-g oral-glucose tolerance test and 24-h dietary recall. The mean fat intake of this population (36% of energy) was typical for North America, but fiber intake (1.2 g/MJ) was very low. Logistic-regression analysis, adjusted for age, sex, and body mass index, showed that a 1-SD increase in fiber intake reduced the risk of having diabetes by 39% (P = 0.026) whereas the same increase in protein intake increased the risk by 38% (P = 0.027). There was no significant effect of energy, fat, starch, or simple sugars. These data support Trowell's original dietary-fiber hypothesis that "... dietary fiber depleted starchy foods are conducive to the development of diabetes mellitus in susceptible human genotypes." PMID- 9394702 TI - Coffee consumption and plasma homocyst(e)ine: results from the Atherosclerosis Risk in Communities Study. PMID- 9394703 TI - Truly quantitative dietary studies have stringent requirements. PMID- 9394704 TI - Vitamin B-12 and folic acid supplementation. PMID- 9394706 TI - Race, ethnicity, and health: how can greater understanding be attained? PMID- 9394705 TI - Unmetabolized folic acid and masking of cobalamin deficiency. PMID- 9394707 TI - Iron stores related to iron absorption. PMID- 9394708 TI - Methodologic issues, theoretical considerations, and design criteria for experimental animal and cell culture experiments. AB - This article provides background information that is important when evaluating the relevance to humans of particular animal or in vitro experiments designed to assess the relations between fatty acids and cancer. Considerations in designing carcinogenesis studies to assess the relation between dietary fatty acids and human cancer include selection of the animal model and design of the experimental diets. Animal carcinogenesis models are generally best for evaluating the early phases of cancer development: the initiation and promotion of cancer. Transplantation protocols have been developed for evaluating the effect of diet on the growth and metastasis of partially or fully transformed cells. The variables that are important in such models are the origin and biology of the cell line, the animal host used for the implantation, the site of transplantation, whether the primary tumor is excised after a period of time to allow for metastasis, and when the diets are fed relative to the different phases of tumor growth and metastasis. Studies in cultured cells have been particularly useful for assessing the mechanisms by which fatty acids affect cancer. Considerations in designing studies with cultured cells include selection of the cell line, cell culture conditions, selection of biological endpoints that are relevant to human cancer, and in vivo confirmation of the mechanisms observed in vitro. Design considerations for each of these experimental approaches are discussed and the contributions of each approach are summarized. PMID- 9394709 TI - Effects of dietary fatty acids on breast and prostate cancers: evidence from in vitro experiments and animal studies. AB - Linoleic acid, an n-6 polyunsaturated fatty acid, is essential for normal mammary tissue development, at least in part because it provides the metabolic precursor required for the biosynthesis of key eicosanoids. A similar requirement applies to the growth of estrogen-independent but apparently not to estrogen-dependent rodent mammary and human breast carcinoma cells in vitro. By way of lipoxygenase products, n-6 fatty acids also regulate expression of the invasive phenotype. High-fat, linoleic acid-rich diets promote chemically induced rat mammary carcinogenesis, virally induced mouse mammary tumor development, and the growth and metastasis of estrogen-independent human breast cancer cells in athymic nude mice. In contrast, saturated fatty acids have no discernible effects on mammary carcinogenesis or progression. Most mechanistic studies have focused on the cyclooxygenase and lipoxygenase products of n-6 fatty acid metabolism, and support is accumulating for interactions between these eicosanoids and growth factors and oncogenes. The investigation of dietary fatty acids in prostate cancer is at an early stage and has been handicapped by a lack of satisfactory animal models. However, there are indications that the n-6 fatty acids perform functions in experimental prostate cancer progression similar to those described for breast cancer. PMID- 9394710 TI - Review of the effects of trans fatty acids, oleic acid, n-3 polyunsaturated fatty acids, and conjugated linoleic acid on mammary carcinogenesis in animals. AB - I review the effects of trans fatty acids, oleic acid, n-3 polyunsaturated fatty acids, and conjugated linoleic acid on mammary carcinogenesis in animals. The goal is not to provide an exhaustive survey of all the publications on these topics; such a Herculean effort has been accomplished by previous reviews, which are cited in the text. Instead, the emphasis is on the consistency or lack of consistency of information regarding each of the above fatty acids, confounding factors that may help to reconcile discrepancies in the database, a perspective of the history of the research, and certain unique or exciting opportunities that are worthy of special attention in evaluations of the relations between specific fatty acids and cancer. This review arrives at four conclusions: 1) There is little evidence that trans fatty acids have an adverse effect on carcinogenesis. 2) The data on cancer protection by oleic acid are not convincing. An inhibitory effect attributed to an increased intake of oleic acid could be due to an inadequate supply of linoleic acid. 3) Although a suppressive response to n-3 polyunsaturated fatty acids is observed in most cases, the availability of linoleic acid is likely to be a confounding factor in determining the final outcome. 4) Conjugated linoleic acid is unique in the sense that concentrations < or = 1% are sufficient for producing significant cancer protection and that this effect seems to be independent of the other fatty acids. PMID- 9394711 TI - Fatty acids and colon cancer in experimental models. AB - Experimental models have several advantages in the study of colon cancer. They can be used to tightly control diet, examine putative intermediate markers, test hypotheses about mechanisms of carcinogenesis, and quantify development of tumors in a short time. Dietary issues that have been studied in animal models but are unresolved include the concept of the effects of total fat compared with energy intake, composition of the basal diet, linoleic acid requirements, and interactions of fat with other nutrients. Intermediate markers that have been probed in animal or in vitro studies include cytokinetics, aberrant crypt foci, eicosanoids and hydroxyoctadecadienoic acids, ornithine decarboxylase, tyrosine kinase, protein kinase C, and gene expression. Colon cancer is studied in animals primarily with use of chemicals that are relatively specific inducers of these tumors, but transplantable models and transgenic animals are also used. Total dietary fat is generally thought to affect colon tumorigenesis, but there does not appear to be any specific fatty acid that promotes the development of colon cancer. Several studies indicate that n-3 fatty acids from marine sources alter a variety of biological intermediates and inhibit colonic tumorigenesis; this is probably mediated via the eicosanoid pathway. Although there are undoubtedly multiple cellular changes elicited by certain fatty acids, our current knowledge of this area suggests that specific fatty acid metabolites or their targets are the likely mediators in this sequence. PMID- 9394712 TI - Animal studies: summary, gaps, and future research. AB - Animal models are essential in cancer research but investigators should recognize the limits of the models they use. Because there is no ideal animal model, researchers should use the biological and biochemical diversity among the models to experimental advantage. The differences can tell us as much as the similarities. Fatty acid metabolism seems to play a role in hormone-dependent and hormone-independent cancers, and cell culture experiments have yielded much information on possible mechanisms. However, a knowledge gap exists between these studies and a full understanding of mechanisms in vivo. Mechanisms must be understood before the possible relevance of the findings to humans can be confidently assessed. There is little evidence to suggest that either trans fatty acids or oleic acid has any specific effect on carcinogenesis and it is unlikely that further study will reveal something important but heretofore overlooked. By contrast, there appear to be notable gaps in our understanding of n-3 fatty acids, linoleic acid, and conjugated linoleic acid in relation to possible effects on cancer in humans. The major knowledge gap, and our greatest challenge, is relating promising data from animal models and cell culture studies to the prevention of cancer in humans. PMID- 9394713 TI - Issues in the design and interpretation of studies of fatty acids and cancer in humans. AB - The methods used in nutritional epidemiology to study the relations between fatty acids and cancer risk include ecologic studies, case-control studies, cohort studies, and intervention trials examining either intermediate markers of cancer risk or cancer incidence. Each type of study design has its particular strengths and weaknesses. The inaccuracy of estimates of fatty acid intake from the use of dietary questionnaires linked to nutrient databases is a major limitation in nutritional epidemiology. Information on the concentrations of fatty acids in the circulation or in adipose tissue can complement estimations of dietary intake. Cancer prevention studies now underway are designed to test whole-diet effects on neoplasia and will not be able to separate the effects of specific fatty acids from those of other nutrients in the diet. The development of better intermediate markers of cancer risk could enable the use of experimental methods to assess the relation between specific fatty acids and cancer. Research findings as described in the literature are complicated both by the multiple hypotheses that can be tested when assessing fatty acid effects and by the uncertainties of multivariate adjustment. Although there are substantial obstacles to understanding the relations between fatty acid intakes and cancer risk, there is no better species than humans for inference about diet and cancer risk in people. PMID- 9394714 TI - Biomarkers of fatty acid exposure and breast cancer risk. AB - Study of the effects of most individual biologically active dietary fatty acids on human disease requires the use of biomarkers of long-term intake in well designed epidemiologic studies. Several small studies of tissue taken from women undergoing surgery for breast abnormalities have compared fatty acid profiles of women with ascertained metastatic breast cancer with those with other abnormalities. These studies, although often flawed in design and generally of inadequate statistical power to determine significant differences, provide some evidence. Human studies are generally consistent with animal models suggesting a protective effect of n-3 fatty acids, a detrimental effect of high n-6 fatty acids, and the possible importance of the ratio of these two classes of dietary fatty acids on both breast cancer incidence and recurrence. High intakes of monounsaturated fatty acid were also often negatively associated with breast cancer. The effects of trans fatty acids have rarely been studied, but there are some indications that they may enhance risk. In general, the study of individual fatty acids is in its infancy. Larger well-designed studies with diverse population and modern analyses of individual fatty acids are needed. PMID- 9394715 TI - Specific fatty acids and risks of breast and prostate cancer: dietary intake. AB - Although international comparisons have suggested positive associations between consumption of total or saturated fat and risk of breast cancer, these relations have not been supported in large prospective studies in which confounding factors were minimized. There is no suggestion from international comparisons, case control, or cohort studies that monounsaturated fat (the most abundant fat in the US diet) increases risk of breast cancer, and there is some evidence that higher intake, particularly in the form of olive oil, might actually reduce risk. The available epidemiologic evidence provides little support for any important relation between intake of either linoleic acid or extra-long-chain n-3 fatty acids from fish and risk of breast cancer. However, high consumption of linoleic acid is a relatively recent phenomenon in Western societies and continued evaluation of its relation with breast cancer risk is warranted because of animal data suggesting possible adverse effects. Ecologic, case-control, and cohort studies all support a positive relation between consumption of animal fat and risk of prostate cancer, but current evidence suggests that vegetable fat is not related to risk of this cancer. Although relevant data are limited, neither linoleic acid nor extra-long-chain n-3 fatty acid consumption appears to be related to risk of prostate cancer. Because of the strong evidence that some aspect of foods high in animal fat increases risk of prostate cancer, further studies of specific dietary fatty acids in relation to the occurrence of this malignancy are likely to be particularly valuable. PMID- 9394716 TI - The role of fat, fatty acids, and total energy intake in the etiology of human colon cancer. AB - A high correlation between national per capita disappearance of fat and national rates of colon cancer led to the hypothesis that consumption of fat, especially from animal sources, increases risk for colon cancer. Over the past two decades, this hypothesis has been tested in numerous case-control and cohort studies. In general, neither case-control nor cohort studies find that the total fat composition of the diet increases risk of colon cancer. Case-control studies frequently find that total energy consumption is related to a higher risk of colon cancer, but this result is difficult to interpret because physical activity appears to be protective whereas obesity increases risk. In contrast with the results for total fat, epidemiologic data regarding the role of specific fatty acids are sparse. Nonetheless, useful information regarding major fatty acids may be inferred from the numerous studies that have examined major source of various fats in relation to colon cancer. Intake of red meat or beef has been related to colon cancer risk in most case-control and cohort studies, whereas dietary fat from sources other than red meat, including dairy, poultry, and vegetable oils, does not increase risk of colon cancer. The apparent influence of red meat does not appear to be mediated through its total lipid content, suggesting that other factors such as heterocyclic amines formed during cooking may be critical. Mechanisms whereby fat or red meat may influence colon carcinogenesis are discussed, although none appear compelling. PMID- 9394717 TI - Bridging animal and human studies: what are the missing segments in dietary fat and prostate cancer? AB - Epidemiologic investigations have suggested an association of dietary fat intake with prostate cancer risk, especially risk of advanced prostate cancer. However, supporting evidence from animal studies is limited. Segments that would bridge animal and human studies on dietary fat and prostate cancer--which would determine the future directions of research--are missing. Such segments include 1) well-designed animal studies to evaluate whether dietary fat or fatty acid modulation, particularly by reducing dietary fat and supplementing n-3 fatty acids, reduces the progression of prostate cancer; 2) in vivo identification of intermediate biomarkers that are responsive to dietary fat and fatty acid treatment and may serve as surrogate endpoints in future clinical studies; 3) elucidation of mechanisms by which dietary fat or fatty acid modulation could prevent prostate cancer progression; 4) further epidemiologic studies to estimate dietary exposure more precisely to establish the correlation between dietary fat and risk of prostate cancer; 5) randomized clinical intervention trials evaluating whether dietary fat reduction combined with dietary n-3 fatty acid supplementation delays the recurrence of prostate cancer and improves survival in patients with clinical disease after therapeutic treatment, and whether it prevents or reduces the progression to clinically significant disease in men with latent disease; and 6) studies validating intermediate biomarkers as surrogate endpoints. PMID- 9394719 TI - It's a jungle out there: biodiversity and the physician-scientist. PMID- 9394718 TI - Human studies on the effects of fatty acids on cancer: summary, gaps, and future research. AB - Both the goal of understanding the basic biology of cancer development as well as the practical considerations involving public health, marketing, and nutrition education have stimulated interest on the effects of individual fatty acids on cancer. Data on diet-disease relations must meet the standard of significant scientific agreement based on the totality of publicly available scientific evidence. The process of arriving at significant scientific agreement begins by collecting data accumulated from epidemiologic associations, animal studies, and clinical trials. This supplement represents one useful effort toward that end. Questions raised include: 1) What stage of the disease is being studied, what are the relevant characteristics of the study participants, how well characterized was the diet, and were appropriate experimental designs used; 2) Is there is a significant role for markers of disease and, if so, are the markers available; 3) Is there consistency, strength, and quality of evidence for establishing basic scientific relations; 4) Do dose-response relations, biological plausibility, and temporal relations (with special attention to clinical trials) exist; 5) What is the level of specificity of the data for the health claim or posited relation; and 6) What are the effective amounts of fatty acids in foods and diet? More complete food-composition data with respect to fatty acids and more comprehensive food tables are needed, as are better methods for measuring fat intakes, better markers of progression, and more definitive epidemiologic and clinical studies. At present there is insufficient evidence to conclude that specific fatty acids are associated with cancer development in humans. PMID- 9394720 TI - Image of the month. Cronkhite-Canada syndrome. PMID- 9394721 TI - Risk and significance of endoscopic/radiological evidence of recurrent Crohn's disease. AB - BACKGROUND & AIMS: The aim of this study was to determine the risk of endoscopic/radiological recurrence of Crohn's disease postoperatively and the long-term outcome. METHODS: A randomized placebo-controlled trial was performed to determine the effectiveness of mesalamine in preventing recurrent Crohn's disease postoperatively. Patients in the control group were examined endoscopically/radiologically before entry into and annually during the trial. Findings were classified as minimal or severe. RESULTS: There were 76 patients (49 men and 37 women; mean age, 37.1 +/- 13.2 years). Fifty (61.7%) had terminal ileal resections. Overall, 55 endoscopic/radiological recurrences were observed in 51 patients (67.1%). Expressed actuarially, the recurrence rate was 27.5% at 1 year (95% confidence interval [CI], 15.8%-37.6%), 60.8% at 2 years (95% CI, 46% 71.3%), and 77.3% at 3 years (95% CI, 62.7%-86.3%). Nineteen (37%) were symptomatic and 12 (24%) were initially asymptomatic but later became symptomatic (mean, 13.0 +/- 8.8 months), whereas 20 (39%) remained asymptomatic (mean, 16.9 +/- 17.4 months). Patients with severe endoscopic/radiological disease were significantly more likely to be or become symptomatic than those with minimal disease (23 of 32 vs. 8 of 19, respectively; P = 0.0437). CONCLUSIONS: This study suggests that postoperative endoscopic/radiological recurrences occur later than previously reported. Furthermore, many of these patients, especially with minimal disease, will remain asymptomatic. PMID- 9394722 TI - Antineutrophil cytoplasmic antibodies in T-cell receptor alpha-deficient mice with chronic colitis. AB - BACKGROUND & AIMS: Antineutrophil cytoplasmic antibodies (ANCAs) have been detected in approximately 60% of sera from patients with ulcerative colitis. The aim of this study was to examine the presence of ANCAs and distribution of ANCA producing B cells in the lymphoid tissue of T-cell receptor alpha-deficient (TCR alpha-/-) mice that develop chronic colitis resembling human ulcerative colitis. METHODS: Sera from 87 TCR-alpha-/- mice were tested for the presence of ANCAs by enzyme-linked immunosorbent assay against a human neutrophil extract and selected antigens and by immunofluorescence study using human neutrophils. Enzyme-linked immunospot assay was used for detecting ANCA-producing cells from spleen, mesenteric lymph node (MLN), or colon. RESULTS: Approximately 70% of sera from TCR-alpha-/- mice showed reactivity against human neutrophil extracts by enzyme linked immunosorbent assay. Sixty percent of ANCA-positive sera showed a perinuclear reaction pattern. By enzyme-linked immunospot, ANCA-producing cells were detected in MLN and colon and less often in the spleen of TCR-alpha-/- mice with chronic colitis. The predominant immunoglobulin isotype of these autoantibodies was immunoglobulin A in the colon but not in the MLN and spleen. CONCLUSIONS: TCR-alpha-/- mice produce ANCAs. The ANCA (immunoglobulin A isotype) producing B cells exist primarily in the diseased colonic mucosa of TCR-alpha-/- mice. PMID- 9394723 TI - Regulation of gastric epithelial cell growth by Helicobacter pylori: offdence for a major role of apoptosis. AB - BACKGROUND & AIMS: Helicobacter pylori may affect the normal balance between gastric epithelial cell proliferation and epithelial cell death, thus interfering with the maintenance of gastric mucosal integrity. The aim of this study was to investigate the effect of H. pylori on cell growth, DNA synthesis, induction of apoptosis, and viability of human gastric epithelial cells in vitro. METHODS: H. pylori was incubated with a differentiated human gastric cancer cell line for up to 72 hours, and the effects on cell numbers (cell counts and WST-1 assay), DNA synthesis (5-bromo-2'-deoxyuridine assay and [3H]thymidine incorporation), and DNA fragmentation (DNA fluorochrome staining, transmission electron microscopy, and histone enzyme-linked immunosorbent assay) were assessed. RESULTS: Incubation of gastric epithelial cells with H. pylori led to a time- and concentration dependent reduction of epithelial cell growth and a concomitant induction of DNA fragmentation. At high bacteria-cell ratios (> 100), inhibition of cell growth was associated with a reduction in DNA synthesis. Treatment of gastric cells with tumor necrosis factor alpha, a receptor-activating CD95/APO-1/Fas antibody, and interferon gamma markedly potentiated H. pylori-induced DNA fragmentation. CONCLUSIONS: H. pylori affects gastric epithelial cell growth by direct induction of apoptosis and inhibition of DNA synthesis and indirectly by sensitization of epithelial cells for apoptosis induced by proinflammatory stimuli. PMID- 9394724 TI - Murine CD4 T-cell response to Helicobacter infection: TH1 cells enhance gastritis and TH2 cells reduce bacterial load. AB - BACKGROUND & AIMS: Previous findings suggest that TH1 cellular immune responses contribute to Helicobacter-associated gastritis. To further investigate this issue, interleukin 4 gene targeted mice were infected with Helicobacter felis, and a series of adoptive transfer experiments was performed to evaluate the role of both TH1 and TH2 cells. METHODS: Antigen-specific spleen cells from immunized/challenged or nonimmunized/infected mice or CD4+ T-cell lines were transferred adoptively into naive recipients before live bacterial challenge. RESULTS: Transfer of cells from both groups of donors as well as TH1 or TH2 cell lines exacerbated gastric inflammation in the recipients. No effect on bacterial load was observed in recipients of bulk spleen cells from infected mice or recipients of TH1 cell lines. In contrast, when either a TH2 cell line or bulk cells from immunized challenged mice were transferred adoptively, recipients showed a dramatic reduction in bacterial load. Increased numbers of bacteria were also noted in interleukin 4-deficient mice. CONCLUSIONS: These data suggest a differential contribution of TH1 and TH2 cell-mediated immune responses in Helicobacter infection: one associated with the pathogenesis of disease (TH1 phenotype) and the other associated with protection from or control of infection (TH2 phenotype). PMID- 9394725 TI - Roles of hepatocyte growth factor and its receptor Met during gastric ulcer healing in rats. AB - BACKGROUND & AIMS: It is unclear which growth factors are primarily responsible for stimulating gastric ulcer healing. The roles of hepatocyte growth factor (HGF) and Met/HGF receptor during gastric ulcer healing were studied in rats. METHODS: HGF and Met/HGF receptor were located and quantified by in situ hybridization and immunohistochemistry during experimental gastric ulcer healing. The in vivo effects of exogenous recombinant human HGF on cell proliferation and ulcer healing were assessed and compared with those of placebo and omeprazole treatment. RESULTS: Compared with intact oxyntic mucosa, messenger RNA (mRNA) of HGF and met was substantially greater in the ulcerated region on days 3 and 15. HGF mRNA was located in stromal cells between the regenerative glands and in the arterial vessels of submucosal tissue, whereas met mRNA was located in the epithelial cells of the regenerative glands. After cryoinjury, immunoreactivity for the Met/HGF receptor was absent on day 3, reappeared on day 8, and was overexpressed on day 15. Exogenous recombinant human HGF had no effect on the ulcer healing parameters over days 3-8, but it did increase epithelial cell proliferation in the ulcer margin over days 8-15. CONCLUSIONS: These data suggest that HGF mediates specific tissue interactions between mesenchyme and epithelia during gastric ulcer healing. PMID- 9394726 TI - Enteropathogenic Escherichia coli-induced myosin light chain phosphorylation alters intestinal epithelial permeability. AB - BACKGROUND & AIMS: Infection of epithelial cells with enteropathogenic Escherichia coli (EPEC) induces phosphorylation of the 20-kilodalton myosin light chain (MLC20). The physiological consequence of this biochemical observation, however, has not been discerned. The aim of this study was to determine if EPEC induced phosphorylation of MLC20 was involved in the associated perturbation of intestinal epithelial barrier function. METHODS: Cultured intestinal epithelial cells, T84, were infected with EPEC. The effects of protein kinase inhibitors on EPEC-induced perturbation of barrier function were assessed using electrophysiological techniques. Alterations in MLC20 phosphorylation were correlated with functional responses. RESULTS: Inhibition of myosin light chain kinase, but not protein kinase C or tyrosine kinase, prevented the decrease in resistance caused by EPEC infection and significantly diminished EPEC-induced MLC20 phosphorylation. Epithelial cell monolayers genetically manipulated to constitutively increase MLC20 phosphorylation were relatively resistant to the effects of EPEC on barrier function. CONCLUSIONS: For the first time, these data show that a physiological consequence of the long-recognized increase in MLC20 phosphorylation by EPEC is perturbation of intestinal epithelial barrier function, which probably contributes to the diarrhea associated with this infection. PMID- 9394727 TI - A selective cyclooxygenase 2 inhibitor suppresses the growth of H-ras-transformed rat intestinal epithelial cells. AB - BACKGROUND & AIMS: Constitutive expression of cyclooxygenase 2 (COX-2) has been found in 85% of colorectal cancers. Ras mutations are found in 50% of colorectal adenocarcinomas. The aim of this study was to determine the role of COX-2 in ras induced transformation in rat intestinal epithelial (RIE) cells. METHODS: Cell growth was determined by cell counts. The expression of COX-2 was examined by Northern and Western analyses. For tumorigenicity assays, cells were inoculated into dorsal subcutaneous tissue of athymic nude mice. DNA-fragmentation assays were performed to detect apoptosis. RESULTS: The expression of COX-2 was increased in RIE-Ras cells at both messenger RNA (9-fold) and protein (12-fold) levels. Prostaglandin I2 levels were elevated 2.15-fold in RIE-Ras cells. Serum deprivation further increased COX-2 expression 3.8-fold in RIE-Ras cells. Treatment with a selective COX-2 antagonist (SC58125) inhibited the growth of RIE Ras cells through inhibition of cell proliferation and by induction of apoptosis. SC-58125 treatment reduced the colony formation in Matrigel by 83.0%. Intraperitoneal administration of SC-58125 suppressed RIE-Ras tumor growth in nude mice by 60.3% in 4 weeks. SC-58125 treatment also induced apoptosis in RIE Ras cells as indicated by increased DNA fragmentation. CONCLUSIONS: Overexpression of COX-2 may contribute to tumorigenicity of ras-transformed intestinal epithelial cells. Selective inhibition of COX-2 activity inhibits growth of ras-transformed intestinal epithelial cells and induces apoptosis. PMID- 9394728 TI - A K-ras oncogene increases resistance to sulindac-induced apoptosis in rat enterocytes. AB - BACKGROUND & AIMS: Mutations of c-K-ras occur commonly in colonic neoplasms. The aim of this study was to determine how c-K-ras mutations alter the responses to the chemopreventive agent sulindac. METHODS: The parental rat intestinal cell line IEC-18 and c-K-ras-transformed derivatives were treated with sulindac sulfide. Cell cycle distribution was determined by flow-cytometric analysis (fluorescence-activated cell sorter), apoptosis by DNA fragmentation (laddering), flow cytometry, and microscopy, and changes in gene expression by immunoblotting. RESULTS: Sulindac sulfide inhibited cell growth and induced apoptosis in a time- and dose-dependent manner more rapidly in and at lower concentrations in parental cells than ras-transformed cells. Expression of the sulindac sulfide arrested cells in G0/G1, but cells entered apoptosis throughout the cell cycle. Proapoptotic protein Bak was relatively high in untreated parental cells and increased markedly after sulindac sulfide but was low in untreated ras transformed cells and did not increase after sulindac sulfide. Expression of other Bcl-2 family members was unchanged after sulindac sulfide. However, sulindac sulfide reduced levels of cyclin D1 protein and cyclin E- and cyclin D1 associated kinase activity. CONCLUSIONS: c-K-ras-transformed enterocytes are relatively resistant to sulindac sulfide-induced growth inhibition and apoptosis, which may result from specific reduction of bak expression. PMID- 9394729 TI - Bile regulates the expression of major histocompatibility complex class II molecules on rat intestinal epithelium. AB - BACKGROUND & AIMS: Major histocompatibility complex (MHC) class II molecules are expressed on intestinal epithelial cells, and the intensity of this expression is regulated. The aim of this study was to test the hypothesis that bile regulates the expression of MHC class II molecules on intestinal epithelium. METHODS: Rats were deprived of intestinal bile by external drainage for 24 or 48 hours, and their intestines were collected, sectioned, and stained with the anti-MHC class II monoclonal antibodies OX4 and OX6. For one group of rats, bile flow was deviated from its usual entry point to the ileum. RESULTS: Compared with intact animals, MHC class II expression was observed to be diminished within 24 hours and totally absent after 48 hours of bile drainage. For the group in which bile flow was deviated to the ileum, staining was only observed in the region distal to the entry point. Analysis by bioassay and enzyme-linked immunosorbent assay of bile showed the presence of tumor necrosis factor and interferon gamma, respectively. CONCLUSIONS: It is concluded that the presence of bile is required for the expression of MHC class II molecules on gut epithelium and that the cytokine components of bile may be the inducing agents. PMID- 9394730 TI - Differential expression of galectin 3 and galectin 1 in colorectal cancer progression. AB - BACKGROUND & AIMS: Galectins are beta-galactoside-binding proteins possibly involved in tumor progression. The aim of this study was to determine the pattern of galectin 3 and galectin 1 expression and involvement in colorectal cancer progression. METHODS: Galectin 3 expression was examined immunohistochemically in 39 samples of normal mucosae, 25 adenomas, 87 carcinomas, and 39 lymph node metastases. Galectin 1 was analyzed in 25 samples of mucosae, 15 adenomas, 25 carcinomas, and 11 metastases. Western blot analysis was also performed. RESULTS: All normal mucosae showed strong nuclear galectin 3 expression, which was down regulated in the neoplastic progression, because only 60% of adenomas, 48% of carcinomas, and 44% of metastases were strongly positive (P < 0.0001). Cytoplasmic expression was down-regulated in adenomas (16%) but increased again in carcinomas (64%) (P < 0.0001). Galectin 1 expression was mainly detected in stromal cells and correlated with tumor progression from normal mucosae to adenomas and carcinomas (P < 0.0001). CONCLUSIONS: Galectin 3 expression is down regulated in the initial stages of neoplastic progression, whereas a dissociated cytoplasmic expression increases in later phases of tumor progression. Galectin 1 in colorectal mucosa is predominantly a stromal product whose overexpression is associated with the neoplastic progression of colorectal cancer. PMID- 9394731 TI - The neurochemical coding and projections of circular muscle motor neurons in the human colon. AB - BACKGROUND & AIMS: Enteric neurons can be characterized by their chemical coding, projections, and morphology. The aim of this study was to describe the different classes of human colonic circular muscle motor neurons. METHODS: Human colonic circular muscle motor neurons were identified by retrograde tracing with 1,1' didodecyl 3,3,3',3'-indocarbocyanine perchlorate (Dil) applied to the circular muscle layer. Whole-mount preparations of the myenteric plexus were then double labeled with antisera to choline acetyltransferase (ChAT) and/or nitric oxide synthase (NOS), or NOS and vasoactive intestinal peptide (VIP), and the position and immunoreactivity of Dil-filled neurons were recorded. RESULTS: Fifty-two percent of all Dil-filled neurons were ChAT immunoreactive, and 86% of these projected up to 11 mm orally, with 14% projecting short distances anally. Forty eight percent of the Dil-filled neurons were NOS immunoreactive, and 77% of these projected up to 19 mm anally, with 23% projecting no more than 6 mm orally. A subpopulation of these NOS-immunoreactive motor neurons were also VIP immunoreactive. A small population of myenteric neurons was immunoreactive for both ChAT and NOS, but none projected to the circular muscle. NOS-immunoreactive motor neurons projected for longer distances than those with ChAT immunoreactivity and were larger. CONCLUSIONS: There are two classes of human colonic motor neurons: one is excitatory (ChAT-immunoreactive) and mainly projects orally and the other is inhibitory (NOS +/- VIP immunoreactive) and projects preferentially anally. PMID- 9394732 TI - Corticosteroids repress the interleukin 1 beta-induced secretion of collagenase in human intestinal smooth muscle cells. AB - BACKGROUND & AIMS: The cytokine interleukin (IL)-1 beta induces collagenase expression and inhibits collagen expression in human intestinal smooth muscle (HISM) cells. Corticosteroids cause transrepression of certain genes, including the collagenase gene. The aim of this study was to determine if corticosteroids repress the induction of collagenase expression and the inhibition of collagen secretion by IL-1 beta in HISM cells. METHODS: HISM cells were exposed to IL-1 beta (1-100 pmol/L) for 24 hours in the presence or absence of dexamethasone (10( 6) mol/L). Collagenase messenger RNA (mRNA) levels were determined by ribonuclease protection assay. Collagenase and tissue inhibitor of metalloproteinase protein secretion were determined by enzyme-linked immunosorbent assay of culture medium. Procollagen and collagen secretion were determined by polyacrylamide slab gel analysis of radiolabeled proteins in culture medium. RESULTS: A 30-fold induction of collagenase mRNA and collagenase protein secretion by IL-1 beta was completely abrogated by dexamethasone. Dexamethasone at 10(-6) mol/L also reduced basal levels of collagenase mRNA by 50% and blocked the IL-1 beta-induced inhibition of collagen secretion. CONCLUSIONS: Corticosteroids repress the collagenolytic action of IL-1 beta on HISM cells in vitro and therefore should promote healing in the inflamed intestine. PMID- 9394733 TI - Recombinant factor VIIa corrects prothrombin time in cirrhotic patients: a preliminary study. AB - BACKGROUND & AIMS: Cirrhotic patients with a prolonged prothrombin time (PT) are known to have low levels of factor VII. Because the current modalities to correct this problem are not ideal, recombinant factor VIIa (rFVIIa) may be useful in correcting the prolonged PT observed in the coagulopathy of cirrhosis. The aim of this study was to evaluate the effectiveness of rFVIIa in nonbleeding volunteer patients with the coagulopathy of cirrhosis. METHODS: A preliminary, single center, dose-escalation trial was performed. Cirrhotic patients with a PT of > 2 seconds above the upper limit of the reference value received an intramuscular injection of vitamin K. Ten patients whose PT did not correct to within 2 seconds above the control of the upper limit of the reference value were given three successive dosages of rFVIIa (5, 20, and 80 micrograms/kg) during a 3-week period. RESULTS: The mean PT transiently corrected to normal in all three dosage groups. No adverse effects were noted. There was no evidence of the induction of disseminated intravascular coagulation. CONCLUSIONS: This preliminary trial shows rFVIIa to be effective in transiently reversing the prolonged PT in a select group of nonbleeding cirrhotic patients. These preliminary observations support conducting a large-scale efficacy trial. PMID- 9394734 TI - Vesicle movement in rat hepatocytes is reduced by ethanol exposure: alterations in microtubule-based motor enzymes. AB - BACKGROUND & AIMS: Ethanol is known to alter vesicle-mediated protein trafficking in hepatocytes by undefined mechanisms. In this study, the effects of long- and short-term ethanol exposure on vesicle movements were measured in isolated hepatocytes, and alterations in the function of the microtubule-associated motor enzymes dynamin, kinesin, and dynein, which are believed to support the transport and/or budding of vesicles along microtubules, were tested. METHODS: Vesicular movements in isolated hepatocytes exposed to short- and long-term ethanol treatment were measured. Motor adenosine triphosphatase activities and their association with specific membrane organelles were assessed in response to long term administration of ethanol in vivo or acetaldehyde in vitro. RESULTS: Hepatocytes exposed to short- or long-term ethanol treatment showed a significant reduction in the number of motile vesicles. No alterations in the levels of motor messenger RNA, protein, or enzymatic activity were observed. Interestingly, ethanol had no effect on the association of dynein and kinesin with membranes, whereas there was a significant increase in the amount of dynamin associated specifically with Golgi membranes. CONCLUSIONS: Long- and short-term ethanol exposure markedly reduces hepatocellular vesicle transport by a mechanism apparently independent of any alteration in the enzymatic activity of molecular motors, possibly involving a change in the function of dynamin. PMID- 9394735 TI - Quantitative estimations of the contribution of different bile acid pathways to total bile acid synthesis in the rat. AB - BACKGROUND & AIMS: Cholesterol degradation to bile acids occurs via "classic" or "alternative" bile acid biosynthetic pathways. The aim of this study was to assess the contributions of these two pathways to total bile acid synthesis in vivo. METHODS: Rats with biliary fistulas were infused with squalestatin for 24 and 48 hours; specific activities of cholesterol 7 alpha-hydroxylase (C7 alpha H) and sterol 27-hydroxylase (S27H) and rates of bile acid synthesis were determined. RESULTS: Continuous squalestatin infusion (15 micrograms/h) decreased C7 alpha H specific activities to 4% and 12% of paired biliary fistula controls at 24 and 48 hours, respectively (P < 0.05) without any changes in S27H specific activities (82% and 95% of controls). At 24 hours, bile acid synthesis decreased to 43% (P < 0.05) but returned to 87% at 48 hours (P = NS). Cholic acid synthesis decreased at 24 hours but returned to control levels at 48 hours. Similar changes in C7 alpha H, S27H, and bile acid synthesis were observed in primary rat hepatocytes after addition of squalestatin (1.0 mumol/L). CONCLUSIONS: In the face of persistent suppression of C7 alpha H and the classic pathway, an alternative pathway becomes a main pathway of bile acid synthesis capable of generating cholic and chenodeoxycholic acids. The observed induction of bile acid synthesis via an alternative pathway or pathways represents an important mechanism for maintenance of cholesterol homeostasis in the rat. PMID- 9394736 TI - Increased bile acid pool inhibits cholesterol 7 alpha-hydroxylase in cholesterol fed rabbits. AB - BACKGROUND & AIMS: Cholesterol feeding unexpectedly inhibits cholesterol 7 alpha hydroxylase in rabbits. The aim of this study was to explore the mechanism. METHODS: Twenty male New Zealand white rabbits were fed regular chow with and without 2% cholesterol for 10 days followed by 7 days of bile drainage. The activities of hepatic cholesterol 7 alpha-hydroxylase and sterol 27-hydroxylase that control bile acid synthesis in classic and alternative pathways were related to the size and composition of bile acid pool. RESULTS: After feeding cholesterol, plasma and hepatic cholesterol concentrations increased, the bile acid pool doubled (from 254 +/- 44 to 533 +/- 51 mg; P < 0.001), cholesterol 7 alpha-hydroxylase activity decreased 68% (P < 0.01), but sterol 27-hydroxylase activity increased 66% (P < 0.05) with increased cholic acid synthesis (P < 0.01). Bile drainage in the cholesterol-fed rabbits depleted the bile acid pool and stimulated down-regulated cholesterol 7 alpha-hydroxylase activity 11.4-fold (P < 0.001), although hepatic cholesterol remained elevated. Hepatic sterol 27 hydroxylase activity was unaffected. CONCLUSIONS: Feeding cholesterol increased hepatic cholesterol and stimulated sterol 27-hydroxylase and alternative bile acid synthesis, which expanded the bile acid pool and inhibited cholesterol 7 alpha-hydroxylase in rabbits. In distinction, hepatic sterol 27-hydroxylase was insensitive to changes in the bile acid pool. PMID- 9394737 TI - Chemokine gene expression in rat pancreatic acinar cells is an early event associated with acute pancreatitis. AB - BACKGROUND & AIMS: The molecular mechanisms underlying pancreatitis are largely unknown. The goal of this study was to identify an early genetic event that correlated with pancreatitis. METHODS: Differential display of messenger RNAs (mRNAs) was conducted on normal pancreas vs. those of animals with secretagogue induced pancreatitis. Northern blots from normal animals and animals with experimental acute pancreatitis were probed with cloned complementary DNAs for chemokines. Pancreatitis was induced with cerulein and by retrograde injection of bile salts. Immunocytochemistry was used to identify the source of chemokine expression. Pyrrolidine dithiocarbamate was tested for effects on chemokine expression and pancreatitis. RESULTS: A differentially amplified band was consistently observed early after cerulein hyperstimulation. This band was identified as a portion of the mob-1 gene, an alpha-chemokine. Northern analysis indicated that mRNAs for mob-1 and another chemokine, mcp-1, were induced after cerulein hyperstimulation in vivo. mob-1 mRNA was also induced by retrograde injection of bile salts and by cerulein in acinar cells in vitro. mob-1 protein was localized to exocrine cells in pancreata of diseased animals. Pyrrolidine dithiocarbamate inhibited both chemokine gene expression and early inflammatory characteristics of pancreatitis. CONCLUSIONS: Chemokines are induced in acinar cells by treatments that induce pancreatitis and may play an important role in the early stages of the disease. PMID- 9394739 TI - Helicobacter pylori infection in the pathogenesis of duodenal ulcer and gastric cancer: a model. PMID- 9394738 TI - A preS mutation isolated from a patient with chronic hepatitis B infection leads to virus retention and misassembly. AB - A preS mutation derived from a patient with chronic hepatitis B virus (HBV) infection who had HBV reinfection with fibrosing cholestatic hepatitis after orthotopic liver transplantation was characterized. Sequence analysis of the HBV genome revealed two deletions and a point mutation in the regulatory CCAAT element of the S promoter. To investigate the particular preS mutation for replication competence and viral assembly in functional experiments, the mutant preS region was introduced into a replication competent HBV plasmid. Functional studies were performed by transfecting this plasmid into hepatoma cells. Analysis of the mutant HBV strain revealed an inverse ratio of S-gene products in comparison to wild-type HBV that leads to intracellular viral retention. An atypical intracellular distribution of HBV proteins and an enhanced nuclear localization of HBV DNA was also detected. Additionally, a major fraction of the extracellular viral particles was malformed. The association of intracellular accumulation of viral proteins with cirrhosis and fibrosing cholestatic hepatitis has been described recently. In this study, we show that the particular preS mutation accounted for the viral retention, which may have contributed to a more progressive form of liver disease found in this HBV-positive patient after liver transplantation. PMID- 9394740 TI - Nonsteroidal anti-inflammatory drugs and colorectal cancer: evolving concepts of their chemopreventive actions. PMID- 9394741 TI - NSAIDs to prevent colorectal cancer: a question of sensitivity. PMID- 9394742 TI - Galectins: multipurpose carbohydrate-binding proteins implicated in tumor biology. PMID- 9394743 TI - Bile salt biosynthesis: an alternate synthetic pathway joins the mainstream. PMID- 9394744 TI - The gatekeeper has many keys: dissecting the function of the APC gene. PMID- 9394745 TI - By their scars ye shall know them. PMID- 9394746 TI - Lansoprazole, H. pylori, and atrophic gastritis. PMID- 9394747 TI - Gastroesophageal reflux after cure of H. pylori infection. PMID- 9394748 TI - H. pylori eradication in gastroesophageal reflux disease. PMID- 9394749 TI - Reflux esophagitis and H. pylori. PMID- 9394750 TI - Therapeutic placebo in ulcerative colitis: fact or fiction? PMID- 9394751 TI - Cost of endoscopy in economic evaluation. PMID- 9394752 TI - Proceedings of the American Digestive Health Foundation International Update Conference on Helicobacter pylori. McLean, Virginia, USA, February 13-16, 1997. PMID- 9394753 TI - How is Helicobacter pylori transmitted? AB - Helicobacter pylori is one of the world's most common pathogens. It colonizes about 60% of the world's population, causes gastritis and peptic ulcer, and is strongly associated with gastric adenocarcinoma and lymphoma. However, most individuals never develop clinical disease. Thirteen years after the culture of H. pylori by Marshall and Warren, we still do not know its major mode of transmission. Childhood represents the major period of acquisition of infection in the third world, but infection is rare in children in the developed world. Possible routes of infection include either oral-oral or fecal-oral, iatrogenic spread with inadvertent use of unsterile pH probes and endoscopes, and vectorial spread by flies. Evidence to support each route of transmission is provided, but there is no predominant route. The only significant reservoir of infection appears to be humans themselves. The organism has been found in some domestic cats and in nonhuman primates, but the opportunities for human interaction with the latter are rare, making infection from this source an unlikely possibility. The organism has the propensity to become a coccoid form. This may represent a persistent form in which H. pylori can exist in the environment, but it has yet to be shown that it can revert to the replicative form. PMID- 9394754 TI - What are the host factors that place an individual at risk for Helicobacter pylori-associated disease? AB - Helicobacter pylori infection is associated with duodenal and gastric ulcer disease, gastric cancer, and gastric mucosa-associated lymphoid tissue lymphoma. Although more than half the world's population harbors H. pylori, only a proportion will develop clinically significant disease. The specific clinical outcome of an individual can be examined as the modulation of host factors by H. pylori infection. Host acid-secretory status and sensitivity to gastrin can be modulated by H. pylori infection. Once H. pylori has established itself in the stomach, virtually everyone develops gastritis, and variations in gastritis patterns have been associated with different gastric acid responses to H. pylori infection. The patterns of gastritis are important because they seem to determine disease outcome. Blood group antigens have been implicated in studies of ulcer disease. Receptors to Lewis antigens in gastric mucosa indicate that host mucosal factors influence H. pylori attachment. Conversely, H. pylori strains express Lewis antigen-like molecules, suggesting an autoimmune component for some H. pylori-associated diseases. HLA genotypes may influence the host response to H. pylori infection, and those of H. pylori-infected individuals have been correlated with histological features. The clinical outcome of H. pylori infection is most likely a result of complex interactions among host, bacterial, and environmental factors. The mechanisms by which these diverse factors influence the pathogenesis of different clinical outcomes remain under investigation. PMID- 9394755 TI - Helicobacter pylori factors associated with disease development. AB - Although certain factors appear to predispose the host to infection by Helicobacter pylori, clearly the bacterium possesses a well-defined battery of virulence factors that allow the organism to: (1) colonize the gastric mucosa (urease, flagella, adhesins, acid-inhibitory protein, iron acquisition proteins, and heat shock proteins); (2) evade host defense (shedding of surface proteins, catalase, superoxide dismutase, and poorly reactive lipopolysaccharide); and (3) damage host tissue (vacuolating cytotoxin, protease, CagA-related factors, inducers of cytokines, and chemotaxins). Together these factors allow H. pylori to persist in the host, establishing a chronic infection. Although many of these virulence factors are produced by all strains of H. pylori, there are also well defined pathogenicity islands (contiguous stretches of chromosomal DNA) present in some strains that encode additional proteins including CagA that potentiate virulence. Strains possessing these "virulence cassettes" are isolated more frequently from patients with the more serious clinical manifestations associated with duodenal ulcer than from patients with gastritis alone or nonulcer dyspepsia. PMID- 9394756 TI - Commentary: Helicobacter pylori transmission, host factors, and bacterial factors. PMID- 9394757 TI - How does Helicobacter pylori cause mucosal damage? Direct mechanisms. AB - Helicobacter pylori-associated gastritis is characterized by an abundant inflammatory response and gastric epithelial cell injury. Adherence of H. pylori to gastric epithelial cells seems to be required for bacterial colonization of the gastric mucosa. Attachment of the bacterium to polarized gastric epithelial cells causes damage to microvilli and stimulates actin polymerization, which is associated with adherence pedestal formation. Studies suggest that H. pylori directly contributes to the injury of gastric epithelial cells by the elaboration of cytotoxic factors. The first toxin identified from H. pylori strains, known as vacuolating cytotoxin, induces vacuole formation in eukaryotic cells. Elaborated enzymes by H. pylori may also contribute directly to epithelial cell injury. Ammonia produced through urease activity may be toxic to gastric epithelial cells. H. pylori protease and lipase degrade gastric mucus and disrupt the phospholipid-rich layer at the apical epithelial cell surface, allowing for cell injury from back diffusion of gastric acid. This cell injury may lead to cell death, believed to result from induction of apoptosis. There are sufficient data to suggest that H. pylori, through direct pathogenic mechanisms, contributes significantly to the gastric mucosal injury associated with this infection, and may enhance the susceptibility of gastric epithelial cells to carcinogenic conversion. PMID- 9394758 TI - How does Helicobacter pylori cause mucosal damage? The inflammatory response. AB - The role for Helicobacter pylori in the pathogenesis of disease provides the conundrum that only a subset of subjects infected with H. pylori will ever develop peptic ulcer or gastric cancer. Thus, variation in strain as well as environmental or host factors converge in the gastroduodenal milieu and control the final outcome of infection. The host immune and inflammatory response is emerging as an important element in the pathogenesis of these gastric diseases. The ideal host response provides protection to clear an infection without causing excessive amounts of inflammation that could compromise the integrity and function of host cells. This review will cover four main questions: (1) What are the mucosal immune/inflammatory responses that confer protection without damaging the host? (2) How do the gastric immune responses during infection with H. pylori differ from this ideal scenario? (3) Do these responses contribute to autoimmune mediated damage to gastric tissue? (4) Can immunomodulation through vaccination enhance protective, nondestructive responses that prevent or treat infection or, at least, attenuate inflammation? PMID- 9394759 TI - How does Helicobacter pylori cause mucosal damage? Its effect on acid and gastrin physiology. AB - Helicobacter pylori infection increases gastric acid secretion in patients with duodenal ulcers but diminishes acid output in patients with gastric cancer and their relatives. Investigation of the basic mechanisms may show how H. pylori causes different diseases in different persons. Infection of the gastric antrum increases gastrin release. Certain cytokines released in H. pylori gastritis, such as tumor necrosis factor alpha and specific products of H. pylori, such as ammonia, release gastrin from G cells and might be responsible. The infection also diminishes mucosal expression of somatostatin. Exposure of canine D cells to tumor necrosis factor alpha in vitro reproduces this effect. These changes in gastrin and somatostatin increase acid secretion and lead to duodenal ulceration. But the acid response depends on the state of the gastric corpus mucosa. The net effect of corpus gastritis is to decrease acid secretion. Specific products of H. pylori inhibit parietal cells. Also, interleukin 1 beta, which is overexpressed in H. pylori gastritis, inhibits both parietal cells and histamine release from enterochromaffin-like cells. H. pylori also promotes gastric atrophy, leading to loss of parietal cells. Factors such as a high-salt diet and a lack of dietary antioxidants, which also increase corpus gastritis and atrophy, may protect against duodenal ulcers by decreasing acid output. However, the resulting increase of intragastric pH may predispose to gastric cancer by allowing other bacteria to persist and produce carcinogens in the stomach. PMID- 9394760 TI - Helicobacter pylori, inflammation, mucosal damage, and apoptosis: pathogenesis and definition of gastric atrophy. AB - Much of what is currently accepted on the natural history of Helicobacter pylori induced gastritis and its relationship with gastric adenocarcinoma rests on the assumption that atrophic gastritis can be correctly identified and reproducibly recognized. Recently, several studies have indicated that pathologists have a low level of agreement on this topic, and the terms "gastric atrophy" and "atrophic gastritis" remain imprecisely defined and, therefore, poorly understood. Furthermore, the genesis and progression of the atrophic changes taking place in the gastric mucosa of some, but not all, subjects infected with H. pylori are incompletely characterized. This review has three aims: (1) to briefly reexamine our current knowledge of the mechanisms involved in the injury and repair of gastric glands; (2) to present a hypothesis on the development of gastric atrophy; and (3) to propose a new, stringent definition of gastric atrophy that may be usefully applied in the clinical research arena. PMID- 9394761 TI - What role does Helicobacter pylori play in gastric cancer? AB - Several lines of evidence support an association between Helicobacter pylori infection and gastric cancer. The natural history of H. pylori-associated gastritis is inexorable progression ultimately leading to gastric atrophy. In general, this process requires between 20 and 40 years to complete. Atrophic gastritis is widely considered to be a precursor lesion of the intestinal type of gastric cancer. Moreover, areas with a high prevalence of H. pylori infection also have a high prevalence of gastric cancer. Strong evidence from three prospective studies shows the risk of gastric cancer to be increased fourfold in H. pylori-positive persons. Several retrospective studies have also confirmed that H. pylori infection is associated with development of gastric cancer, especially in the younger generation, early gastric cancer, and noncardiac gastric cancer. H. pylori alone is not likely responsible for gastric cancer. Rather, it may provide a suitable environment, including chronic gastritis and intestinal metaplasia, for neoplastic change. Recognition of an association between H. pylori infection and gastric cancer has led to a major shift in emphasis on the cause of the disease. Research into H. pylori has focused attention on the importance of chronic inflammation and impaired host defense mechanisms as factors in the development of gastric cancer. H. pylori infection leads to changes in many factors that are important to the pathogenesis of gastric cancer, including vitamin C content of gastric juice, reactive oxygen metabolites, and epithelial cell proliferation. Eradication of the organism may reverse these changes. Therefore, eradication of H. pylori in infected persons might be a route to preventing gastric cancer, although many questions still remain as to the effectiveness of this strategy. PMID- 9394762 TI - What role does Helicobacter pylori eradication play in gastric MALT and gastric MALT lymphoma? AB - The concept of mucosa-associated lymphoid tissue (MALT) has been introduced to differentiate biological functions from behavior of nonnodal vs. nodal lymphoid tissues. Lymphomas arising from MALT also behave differently than typical nodal lymphomas. In contrast to other tissues, MALT in the stomach is almost exclusively a result of Helicobacter pylori infection. Thus, MALT is part of the host defense against the pathogen H. pylori. Consequently, lymphomas arising from gastric MALT may be a clonal evolution starting from the infection. In low-grade gastric MALT lymphoma, cure of the infection may induce complete histological remission in the majority of patients. Investigators have recently reported that complete remission rate is between 70% and 80%. In an extended analysis, we have treated 84 patients with low-grade gastric MALT lymphoma in stage El, using a dual regimen to eradicate H. pylori. Complete remission was observed in 68 (80%) patients; a partial remission was found in 4 patients. In contrast, 12 patients showed no change and were referred to alternative treatment. In patients in complete remission, a polymerase chain reaction assay for the rearranged immunoglobulin heavy-chain gene remained positive in many cases. Together with data from the literature, these data suggest that the majority of patients with low-grade gastric MALT lymphomas in stage El respond to eradication of H. pylori. Longer follow-up investigations are necessary to determine if remissions indicate a cure from the disease. PMID- 9394763 TI - Commentary: role of Helicobacter pylori on gastric mucosal damage, gastric cancer, and gastric MALT lymphoma. PMID- 9394764 TI - What role does Helicobacter pylori play in dyspepsia and nonulcer dyspepsia? Arguments for and against H. pylori being associated with dyspeptic symptoms. AB - A major role for Helicobacter pylori gastritis in nonulcer dyspepsia (NUD) is controversial. Gastroduodenal dysfunction may be associated with H. pylori infection, but there is little evidence for a causal link with dyspepsia. Population-based studies with appropriate methodology have generally failed to confirm an association between H. pylori and NUD. Furthermore, no definite association between subgroups of NUD (ulcer-like, dysmotility-like, reflux-like, and nonspecific) and H. pylori has been identified however the subgroups have been defined, and no specific symptom pattern characterizes patients with H. pylori infection. Whether H. pylori-induced alterations of gastric physiology can explain NUD remains open to debate while we await the results of more specific experiments. Although acid secretion in response to gastrin-releasing peptide may be increased in a subset of NUD patients who are infected with H. pylori, uninfected patients with NUD have not been assessed and the results require confirmation. Most studies suggest no association between H. pylori and gastroduodenal motor or sensory dysfunction in NUD. Treatment trials have been unconvincing. The trials with bismuth therapy have not been adequately blinded. Furthermore, some studies suggest that H. pylori-negative patients with NUD may respond to bismuth treatment, although the results have not been uniform. Therapies aimed at curing H. pylori infection have produced mixed results, with small positive and negative trials. The trials that have used adequate outcome measures have more often than not been negative. Based on current evidence, H. pylori is not established to be of causal importance in NUD. PMID- 9394765 TI - What is the role of Helicobacter pylori in complicated ulcer disease? AB - The role of Helicobacter pylori in the pathogenesis of duodenal and gastric ulcer and ulcer recurrence is widely known. Bleeding, perforation, and obstruction represent the most serious, potentially life-threatening manifestations of ulcer disease (hemorrhage in 15%-20%). The lifetime prevalence of perforation and obstruction is much lower (approximately 5% and 2%, respectively). Despite improved diagnostic and therapeutic options, bleeding-related mortality rates remain at 6%-7% in the United States and between 4% and 14% in Europe. H. pylori and nonsteroidal anti-inflammatory drugs are now recognized as the two primary causes of ulcer disease. Eradication of H. pylori in patients with uncomplicated ulcers results in recurrence rates of < 10%, suggesting that eradication of H. pylori in patients with bleeding ulcers may virtually prevent recurrence of both the disease and its complications. Although the prevalence of H. pylori infection is almost 100% in duodenal and 80%-90% in gastric ulcer patients not using nonsteroidal anti-inflammatory drugs, the prevalence of the organism in bleeding duodenal and gastric ulcers does not reach 70% and 60%, respectively, which may be due to false-negative test results. PMID- 9394766 TI - Relationship between nonsteroidal anti-inflammatory drug use, Helicobacter pylori, and gastroduodenal mucosal injury. AB - With the realization that Helicobacter pylori is the main etiologic factor for peptic ulcer disease, recent studies have explored a potential relationship between H. pylori and nonsteroidal anti-inflammatory drug (NSAID)-related gastroduodenal mucosal injury. Using serology and/or histology to detect H. pylori, case-control studies have shown no meaningful differences in H. pylori prevalence in both arthritis and nonarthritis NSAID users and controls. Placebo controlled short-term trials of NSAIDs have also shown no change in the frequency of detection of H. pylori by gastric mucosal biopsy specimens after 7-30 days of NSAID ingestion. A number of studies have shown that the histological gastritis identified in NSAID users is caused by H. pylori infection, whereas the reactive (chemical) gastritis can be caused by NSAID use. Although the overall relationship between H. pylori gastritis and dyspepsia remains controversial, there is no evidence from well-controlled studies using either serology or histology that this gastritis predisposes to NSAID-related dyspepsia. The effect of H. pylori on NSAID-related gastroduodenal mucosal injury may be best established by evaluating the ulcer recurrence rate after H. pylori eradication and rechallenge with NSAIDs. To date, only one such study has examined this question, and in this small study, the ulcer recurrence rate at 6 months was not reduced by H. pylori eradication. PMID- 9394767 TI - Commentary: bleeding ulcers, interaction between NSAIDs and Helicobacter pylori infection, and nonulcer dyspepsia. PMID- 9394768 TI - How should Helicobacter pylori infection be diagnosed? AB - The ideal approach for the initial diagnosis of Helicobacter pylori infection is to perform an endoscopy to obtain biopsy specimens for histology and culture. Histology allows classification of any gastritis lesions present and may have prognostic value, and culture enables susceptibility testing of antimicrobial agents to direct proper treatment. Biopsy specimens must also be taken from the corpus if the patient was pretreated with proton pump inhibitors. The cost of these tests and the delay in receiving results limits their use in clinical practice. Therefore, the urease test, a quick and inexpensive test, is used to detect the presence of H. pylori and constitutes the basic invasive test for H. pylori. A new urease test based on a strip instead of an agar disk may be the test of choice in the future, because of its increased sensitivity and 2-hour delay (instead of 24 hours) in obtaining the result. In some countries, because of the cost, endoscopy will be used in selected patients only, either because of alarm symptoms or age > 45 years, which is considered a threshold for gastric cancer risk. In other patients, the noninvasive tests will be used. The cost of serology makes it more attractive compared with the urea breath test. Currently, there are accurate enzyme-linked immunosorbent assay tests that can be performed in any laboratory and that provide precise and quick diagnoses. In the event of a doubtful result, an immunoblot can be performed, as is the case for other infections. Patient follow-up after treatment provides a different situation because bacterial load is usually lower. A noninvasive test should be performed, and only the urea breath test can be used within the timing originally proposed to test eradication efficacy (i.e., 4-6 weeks after treatment). If the result is positive, susceptibility testing is required before administering a second course of treatment. The increasing use of antimicrobial agents to treat H. pylori is likely to result in antimicrobial resistance, requiring that bacteriologic surveillance programs be implemented. There are numerous research projects ongoing in this area, and one can expect that improved methods, such as colorimetric polymerase chain reaction (PCR) and improved antibody tests, will also be used in the future. PMID- 9394770 TI - For what conditions is there evidence-based justification for treatment of Helicobacter pylori infection? AB - Evidence-based medicine combines clinical expertise and the best available evidence from systematic research to aid decision making in patient care. Levels of evidence can be graded from I to V, with level I, the strongest, coming from large randomized controlled trials (RCTs). When a definitive RCT has not been performed, or is impracticable or inappropriate, lesser grades of evidence are used. There is level I evidence supporting the treatment of Helicobacter pylori infection in patients with duodenal or gastric ulcers. Prospective RCTs have shown that cure of the infection is associated with ultimate cure of the ulcer diathesis. Therefore, this is a "grade A" recommendation for treatment. In nonulcer dyspepsia, numerous RCTs have yielded conflicting results regarding the benefits of treatment. Although there are methodological problems with many reported studies, there is some evidence (level II at best) to support treatment- a grade B recommendation. In early gastric cancer and gastric mucosa-associated lymphoid tissue lymphoma, the best available evidence supporting treatment of H. pylori infection is of low quality, i.e., levels III and V. Although these carry only grade C treatment recommendations, treatment is safe and carries at least some evidence of efficacy. It is therefore indicated based on the current best available evidence. No evidence exists to support treating the infection in patients receiving long-term proton pump inhibitors for gastroesophageal reflux disease or in patients with any of the nongastrointestinal conditions that have been tentatively linked to H. pylori. PMID- 9394769 TI - Who should be treated for Helicobacter pylori infection? A review of consensus conferences and guidelines. AB - The publication of the National Institutes of Health Consensus Development Conference guidelines on management of Helicobacter pylori infection in 1994 set a precedence. At present, at least eight European countries have produced national guidelines, and, more recently, the European Helicobacter pylori Study Group also outlined guidelines based on the strength of available evidence. It is generally agreed that H. pylori should be eradicated in peptic ulcer disease. In nonsteroidal anti-inflammatory drug (NSAID)-related ulcers, most countries that considered the issue suggested discontinuing NSAIDs when possible and eradicating H. pylori. The prophylactic eradication of H. pylori was not recommended. A number of panels felt that there was not enough evidence available to recommend eradication of H. pylori in functional dyspepsia, whereas other groups felt that nonulcer dyspepsia, particularly after investigation and with severe or recurrent symptoms, was an indication for eradication therapy. Other conditions (i.e., gastroesophageal reflux disease [GERD] and mucosa-associated lymphoid tissue [MALT] lymphoma) have emerged in this short time as possible indications for H. pylori eradication. There is no evidence that H. pylori infection has a role in the pathogenesis of GERD, but there is evidence suggesting that patients with H. pylori infection who require long-term acid suppression may be at risk of developing atrophic gastritis. The European Helicobacter pylori Study Group has suggested that eradication therapy should be offered to infected family members of patients with gastric cancer. It also recommended that eradication therapy was "strongly recommended" on the basis of "supportive" evidence in gastritis with severe abnormalities and after early resection of early gastric cancer. An "uncertain" recommendation with "equivocal" evidence was given for asymptomatic subjects, extra-alimentary tract disease, the prevention of gastric cancer in the absence of risk factors, and in pediatric patients with recurrent abdominal pain. Despite considerable advances, further research studies are needed to provide definite direction for the treatment of many conditions. PMID- 9394771 TI - Can therapy even be denied for Helicobacter pylori infection? AB - Helicobacter pylori infection is a transmissible bacterial infection of the gastric mucosal surface that causes progressive damage with eventual destruction of the stomach. In the United States, the presence of H. pylori infection in patients carries a lifetime risk of developing peptic ulcer of at least 16% and a 1%-3% risk of developing gastric cancer. An infected individual is also a risk to the community because the infection can be transmitted. A review of the data shows that H. pylori is the only treatable infectious disease with such a high rate of morbidity and mortality that is not the subject of an all-out program to eradicate it from the population. The risk of a serious outcome of untreated asymptomatic H. pylori infection is great, or greater, than with asymptomatic syphilis or tuberculosis. H. pylori infection is a serious public health problem, and thus the presence of H. pylori infection justifies treatment. The question is not whom to treat, but whom to test. The gastroenterology community appears to have been unduly influenced by the fact that H. pylori infection is widespread and often asymptomatic, as well as by the costs and complications of current treatment. H. pylori infection is a serious, worldwide infectious disease with tremendous and unacceptable morbidity and mortality. Although there are no emotional reasons to treat H. pylori infection, there are logical and persuasive scientific reasons to treat. If the tools are available, screening the population for the presence of H. pylori infection with the goal of preventing all H. pylori related diseases is recommended. Our goal should be to totally eliminate H. pylori from the face of the earth, just as we eliminated smallpox. PMID- 9394772 TI - Commentary: how, in whom, and when to diagnose Helicobacter pylori. PMID- 9394773 TI - What are the treatment goals for Helicobacter pylori infection? AB - The goals of therapy of Helicobacter pylori are commonly viewed as either clinical (e.g., avoid further morbidity, mortality, or prevent disease from occurring) or economic (e.g., save healthcare dollars by avoiding further expenditure of resources). However, when viewed globally, these goals are not incongruent but are inexorably linked. Prevention of disease or avoidance of further morbidity and mortality by curing H. pylori infection is obviously an improved clinical outcome, but it also concurrently avoids further utilization of healthcare resources and is "cost-effective," resulting in improved economic outcome as well. The more clinically effective an H. pylori treatment is, the more economically effective it is as well. When comparing regimens, the cost effectiveness is determined by efficacy of the regimen, not its cost. Put more simply, "The most expensive therapy is the one that doesn't work!" Therefore, the goals of the therapy are simple: avoid further morbidity, mortality, and prevent disease while minimizing further utilization of healthcare resources, thus saving money. The most clinically effective therapy is also the most cost-effective therapy. Regimens used should have demonstrated greater than 90% efficacy or effectiveness in the population being treated for H. pylori infection. The regimens should be simple and well tolerated. It is clinically and economically inappropriate to use a regimen not meeting these criteria. PMID- 9394774 TI - Current FDA-approved treatments for Helicobacter pylori and the FDA approval process. AB - U.S. Food and Drug Administration (FDA) approval of new drugs expands treatment options and serves as a "safety net" of well-documented efficacy and safety. The information provided in the package insert facilitates physician education and provides some assurance that marketing information is accurate. As of February 1997, three Helicobacter pylori regimes have been FDA-approved for eradication of H. pylori in infected patients with active duodenal ulcers. Regimen 1, omeprazole + clarithromycin (O/C), was supported by two multicenter, controlled studies with a 6-month follow-up. Eradication rates were 74% (n = 53; 95% confidence interval [CI], 62-85) and 64% (n = 61; 95% CI, 52-76). Twenty-five of 26 patients with failed eradication therapy who were taking O/C with clarithromycin-susceptible strains before treatment and who had pretreatment and posttreatment susceptibility tests performed developed clarithromycin resistance after treatment. Regimen 2, ranitidine-bismuth-citrate + clarithromycin, was supported by two multicenter, placebo-controlled studies with a 6-month follow-up. Eradication rates were 84% (n = 19; 95% CI, 60-96) and 73% (n = 22; 95% CI, 50 88). Insufficient pretreatment and posttreatment susceptibility data were collected to assess antimicrobial resistance. Regimen 3, bismuth subsalicylate + metronidazole + tetracycline + an H2-receptor antagonist, was supported by two pivotal literature-based studies. Eradication rates in patients with duodenal ulcer were 82% (n = 51; 95% CI, 70-92) and 77% (n = 39; 95% CI, 61-89), respectively. When extrapolating the results of these three FDA-approved regimens to the clinical setting, particular aspects of the clinical trial should be kept in mind. These include the type of controls, primary end points used, population studied, and number and type of dropouts. PMID- 9394775 TI - What other regimens are under investigation to treat Helicobacter pylori infection? AB - The most common infection in the world, Helicobacter pylori infection, is very specific, and present experience in treating infectious diseases is not applicable in general for this infection. Animal models (e.g., mouse and ferret) are thus far inadequate as reliable screening models. Old-fashioned trial-and error treatment of infected humans is still the screening model and the gold standard in the evaluation of regimens aimed at eradication of H. pylori. A variety of studies on treatment of H. pylori infection have been performed with varying results. This pooled analysis of the following therapeutic combinations: proton pump inhibitor (PPI) plus two antibiotics or antimicrobials, quadruple therapies, and nonantibiotic regimens is an attempt to make a fair comparison of tested therapeutic strategies aimed at eradicating H. pylori. Data from treatment groups including specified drug combinations are pooled, regardless of dose or duration. Search methods are: MEDLINE 1984-1996, Digestive Disease Week 1988 1996, United European Gastroenterology Week 1992-1996, European Helicobacter pylori Study Group 1988-1996, Asia Pacific Congress 1996, H. pylori International Workshop Hong Kong 1996, and miscellaneous. Eradication rates (efficacy) are presented as intention-to-treat data (i.e., worst-case analysis). Separate subanalyses with regard to study quality, dose, and duration are performed for some groups. A general cost-efficacy analysis is performed based on pooled efficacy data. Convenience data are presented as total number of tablets, total number of intake occasions, and duration of therapy. Drugs evaluated in the analysis are bismuthdicitrate, tetracycline, amoxicillin, nitroimidazoles, macrolides, H2-receptor antagonists, PPIs, sucralfate, and sofalcone. The most effective and convenient drug combinations are the PPI-based triple therapies. No significant difference was observed between the three PPIs. The cure rate did not improve after addition of bismuth. Cost-effectiveness is closely associated with efficacy. PMID- 9394776 TI - What is the role for vaccination in Helicobacter pylori? AB - Helicobacter pylori has been implicated in the etiology of peptic ulcer disease, chronic gastritis, gastric carcinoma, and gastric mucosa-associated lymphoid tissue lymphoma. Although significant progress has been made in treating this infection with combinations of either antimicrobial agents or antimicrobial agents plus proton pump inhibitors, these antimicrobial-based treatments continue to be suboptimal. Over the past few years it has become increasingly recognized that direct mucosal immunization can induce protection from infection at mucosal surfaces. Therefore, prevention of H. pylori infection by oral immunization is an alternative approach for the control of H. pylori disease. Using the Helicobacter felis mouse model or H. pylori mouse model, both prophylactic and therapeutic oral immunizations have been shown to be effective against H. pylori. In addition, several H. pylori proteins have been identified as potential candidate vaccines, and a phase 1 clinical trial has been completed that demonstrates the safety and tolerability of urease as a vaccine antigen. Such antigens in combination with a safe mucosal adjuvant could be used in the form of an oral vaccine administered during childhood before exposure to H. pylori to prevent infection. In addition, therapeutic immunization alone or as an adjunct to antimicrobial therapy may be capable of achieving a cure rate approaching 100%. PMID- 9394777 TI - Commentary: treatment of Helicobacter pylori. PMID- 9394778 TI - What remaining questions regarding Helicobacter pylori and associated diseases should be addressed by future research? View from the Far East. AB - Based on the findings of several epidemiological studies, it is believed that Helicobacter pylori infection is closely associated with gastric cancer. Because some abnormalities, such as severe inflammation in the gastric mucosa, impaired secretion of vitamin C, and increased gastric cell proliferation, improve after cure of the infection, anti-H. pylori therapy may reduce the incidence of gastric cancer. In Japan, the odds ratios for the development of atrophic gastritis and gastric cancer in H. pylori-positive patients are not as high as those reported in Europe and the United States. These findings suggest that factors other than H. pylori may exert a considerable influence on the development of atrophic gastritis and gastric cancer in Japan. It is not known whether atrophy and intestinal metaplasia, possible precursors of gastric cancer, are reversible. H. pylori infection is associated with a "gastritis, intestinal metaplasia, and gastric cancer sequence," but it remains obscure whether the infection is directly associated with the development of gastric cancer. We do not know which age group of patients should be given anti-H. pylori therapy for the prevention of gastric cancer. To elucidate whether the eradication of H. pylori can prevent gastric cancer, a Japanese intervention trial is now in progress. PMID- 9394779 TI - What remaining questions regarding Helicobacter pylori and associated diseases should be addressed by future research? View from Europe. AB - A variety of questions regarding Helicobacter pylori need to be addressed by future research. Further investigations are needed on the relationship between H. pylori and gastric cancer. In particular, the mechanism of the interaction between H. pylori infection and host genetic factors and dietary factors that lead to the cancer need to be unraveled. Also, the reversibility of cancer associated abnormalities (e.g., hypochlorhydria, atrophy, and intestinal metaplasia) by eradication of H. pylori needs to be determined. Noninvasive means of identifying H. pylori-positive subjects at high risk of developing gastric cancer are required for such subjects to be targeted for eradication therapy. Further studies are also required on the interactions between H. pylori and proton pump inhibitor therapy that might predispose to cancer. There is considerable interest in the possibility of noninvasive H. pylori testing replacing endoscopy in determining management of nonelderly patients with uncomplicated dyspepsia unassociated with nonsteroidal anti-inflammatory drugs (NSAIDs). Randomized studies comparing endoscopy vs. noninvasive H. pylori testing in this situation are required with comprehensive outcome measures. Improvement in eradication therapy is required and will depend on the development of more effective and specific antibiotics and therapeutic vaccines. Wide-scale elimination of the infection will depend on preventing its spread from person to person. Achieving this will require further knowledge of its mode of transmission, particularly in childhood, and the development of prophylactic vaccines. Further studies are required to define the role of H. pylori infection in other diseases, including predisposition to enteric infection in the developing world as a result of H. pylori-induced chronic hypochlorhydria, nonulcer dyspepsia, pernicious anemia, atherosclerosis, and NSAID-related ulcer disease. Finally, we need to know whether H. pylori infection may be beneficial in certain circumstances and whether eradicating the infection may be disadvantageous to some subjects. PMID- 9394780 TI - What remaining questions regarding Helicobacter pylori and associated diseases should be addressed by future research? View from North America. AB - Several areas regarding Helicobacter pylori that need improvement or clarification in the United States include treatment of dyspepsia, physician education on disease associations with H. pylori, and evidence from U.S. studies that 7-day H. pylori eradication regimens are more effective than current regimens. Dyspepsia, a ubiquitous condition in the United States, is routinely managed on the basis of a positive H. pylori serology without other investigations. This approach has been fostered by cost-effectiveness studies of various approaches to duodenal ulcer and dyspeptic patients. Serology-directed therapy was the most cost-effective option vs. endoscopy-directed management. The option of not obtaining endoscopy had broad appeal to primary care physicians. In addition, a recent survey suggests that even gastroenterologists routinely attempt H. pylori eradication in infected patients with nonulcer dyspepsia, despite a number of negative efficacy studies. Finally, the option of not eradicating a World Health Organization-defined carcinogen in the litigious United States is unappealing to clinicians. Eradication of H. pylori in patients with dyspepsia despite more negative trials is likely to continue. There is evidence that U.S. physician awareness of the H. pylori-disease associations and the best therapies are improving rapidly, but further improvement is needed. Discrepancy of awareness of H. pylori between gastroenterologists and family physicians exists. In a recent survey, 94% and 72% of gastroenterologists regarded H. pylori as a causative agent in duodenal and gastric ulcer, respectively, vs. 68% and 68% of family physicians, and only 9% of family physicians believed there was a definite relationship between H. pylori infection and gastric cancer vs. 21% of gastroenterologists. One hundred three different H. pylori regimens were being used; 31% of family physicians and 11% of gastroenterologists used ineffective regimens or regimens of unknown effectiveness. Although 1-week proton pump inhibitor triple therapy is promising, there is skepticism that U.S. studies will yield the optimistic results that have characterized the European studies. Unlike in Europe, the U.S. standard is to use double diagnostics to prove eradication rather than just the urea breath test and to use intent-to-treat rather than assessable patient analyses. Both approaches reduce apparent eradication rates. PMID- 9394781 TI - Commentary: Helicobacter pylori and future research. PMID- 9394782 TI - Conditioned immunomodulation: investigations of the role of endogenous activity at mu, kappa, and delta opioid receptor subtypes. AB - The present investigations were designed to determine the role of activity at mu, kappa, and delta opioid receptor subtypes in conditioned immunomodulation by evaluating the effects of selective opioid receptor antagonists on conditioned stimulus-induced alterations in immune status. Lewis rats were exposed to an aversive conditioned stimulus that was developed through pairings with electric footshock. This aversive conditioned stimulus induces a reduction in splenic natural killer cell activity, splenocyte proliferation in response to mitogens, and diminished levels of interferon-gamma (IFN-gamma) production by splenocytes. Intracerebroventricular (i.c.v.) administration of the opioid antagonist naltrexone or the mu 1-selective antagonist naloxonazine blocked conditioned alterations of immune status, indicating that activity at mu-opioid receptors is involved in conditioned immunomodulation. Further support for the involvement of mu-opioid receptors within the central nervous system is provided by data showing that peripheral administration of naloxonazine, at doses shown to be effective when administered i.c.v., had no effect on conditioned alterations of immune status. Ventricular administration of the kappa receptor antagonist nor binaltorphimine (nor-BNI) did not antagonize the immunomodulatory effects of the conditioned stimulus. Administration of the delta receptor antagonist naltrindole also did not antagonize the conditioned alterations of immune status. Collectively, the results of this study indicate that the alterations of immune status produced by an aversive conditioned stimulus require activity at mu-opioid receptors, possibly mu 1, within the central nervous system. PMID- 9394783 TI - Treatment with intact anti-B7-1 mAb during disease remission enhances epitope spreading and exacerbates relapses in R-EAE. AB - PLP139-151-induced experimental autoimmune encephalomyelitis in the SJL mouse is a Th1-mediated inflammatory demyelinating disease characterized by a relapsing remitting clinical course (R-EAE). Clinical relapses are mediated by T cells specific for a non-cross reactive secondary PLP epitope (PLP178-191) induced by epitope spreading. We have previously shown that B7-1 expression is upregulated in SJL mice undergoing R-EAE and in vivo treatment during remission with F(ab) fragments of anti-B7-1 mAb, blocked epitope spreading and disease progression. In contrast, the present study shows that treatment with intact anti-B7-1 mAb exacerbated clinical disease relapses and enhanced CNS demyelination. Anti-B7-1 treated mice showed enhanced in vivo delayed-type hypersensitivity (DTH) to the relapse-associated PLP178-191 epitope and responses to the immunodominant MBP84 104 epitope which are absent in the controls. Thus, ligation of B7-1 by intact mAbs has effects opposite to those of anti-B7-1 F(ab) fragments suggesting that the mAb is directly signaling through B7-1 expressed on T cells and/or APCs. PMID- 9394784 TI - Inhibition of allergic encephalomyelitis in marmosets by vaccination with recombinant vaccinia virus encoding for myelin basic protein. AB - A primary demyelinating form of experimental allergic encephalomyelitis (EAE) resembling human multiple sclerosis (MS) occurs in Callithrix jacchus marmosets following immunization with human white matter. Participation of a T-cell immune response against myelin basic protein (MBP) in this disease model is supported by observations of increased reactivity against MBP in PBMC and of adoptive transfer of an inflammatory form of EAE by MBP-reactive T-cells. To evaluate the effects of ectopic presentation of MBP on marmoset EAE, animals were vaccinated prior to induction of EAE by subcutaneous injection of attenuated strains of vaccinia virus genetically engineered to contain either the entire coding sequence for human MBP (vT15) or the equine herpes virus glycoprotein gH gene (vAbT249). Vaccination with vT15 was followed by transient cytoplasmic and surface membrane expression of MBP in circulating PBMC (15-45 days). The onset of clinical EAE after immunization (pi) was markedly delayed in vT15-vaccinated animals (37-97 days pi, n = 4) compared to vAbT249-vaccinated controls (14-18 days pi, n = 3). Proliferative responses against MBP but not against vaccinia antigens or phytohemagglutinin were suppressed in protected animals. Thus, development of attenuated live viruses carrying genes for myelin antigens could be useful for induction of immunologic tolerance and for modulation of autoimmune demyelination. PMID- 9394785 TI - Modulation of experimental autoimmune neuritis in Lewis rats by oral application of myelin antigens. AB - Experimental autoimmune neuritis (EAN) in Lewis rats is a T cell-mediated disease and serves as an animal model of human inflammatory demyelinating neuropathies. EAN can be induced by immunization with complete bovine peripheral nerve myelin (BPM), the myelin protein P2 or its neuritogenic peptide, each emulsified in complete Freund's adjuvant (CFA). The present study evaluates the effect of oral tolerization with BPM or P2 protein on the development of actively induced EAN. Oral administration of BPM strongly suppressed clinical and histological signs of EAN subsequently induced by BPM/CFA, but feeding of P2 protein alone did not affect its course. In contrast, feeding of BPM did not mitigate the course of EAN subsequently induced by immunization with neuritogenic P2 peptide/CFA. Oral therapy with BPM after onset of myelin-induced EAN only slightly ameliorated the further course of disease, but significantly reduced lethality of this severe form of disease. The findings suggest that immunogenicity of the antigens fed determine strength of tolerance, that downregulation of EAN occurs at the site of immunization and not in the nerve, and that active suppression rather than specific anergization is operative in mediating resistance to EAN. However, only partial tolerance to myelin-induced EAN was achieved in naive animals by transfer of spleen/LN cells from rats orally tolerized with BPM. Although methodic factors may have limited the effect of the cells, the result is suggestive of some contribution of anergy to oral tolerance in the present model. Cholera toxin and LPS were identified as oral adjuvants for BPM and prolonged the state of tolerance. However, LPS exhibited proinflammatory properties if EAN was induced early after BPM/LPS-feeding. Thus, oral application of a mixture of myelin components in combination with cholera toxin may be a useful treatment for chronic inflammatory neuropathies considered autoimmune in nature. PMID- 9394786 TI - Synergism between sirolimus and 1,25-dihydroxyvitamin D3 in vitro and in vivo. AB - The active form of vitamin D, 1 alpha, 25-(OH)2D3, displays immunomodulatory effects in vitro and in vivo at pharmacological levels. We evaluated the dose effect relationship of 1,25(OH)2D3 and sirolimus (rapamycin, RAP) in vitro, on the inhibition of PHA-stimulated PBMC proliferation, by using the median effect analysis. Pharmacological concentrations of 1,25(OH)2D3 (between 10(-9) and 3 x 10(-6) M) interacted synergistically with RAP (combination index value of 0.01 for 50% suppression of PBMC proliferation). In vivo, the effect of 1,25(OH)2D3 and RAP combinations on the evolution of experimental allergic encephalomyelitis in SJL mice was analyzed. 1,25(OH)2D3, given i.p., in monotherapy, at a dose of 2 micrograms/kg every two days, from day -3 until day +19 after disease induction, or RAP, injected daily at a dose of 0.3 mg/kg for the same period, decreased EAE incidence (paralysis in 70 and 55% of the animals, respectively, versus 98% in the placebo treated group, p < 0.001). The combination treatment using the two drugs in these subtherapeutical doses provided near-total clinical (8% paralysis, p < 0.001 compared to monotherapy with 1,25(OH)2D3 or RAP) and histological protection, comparable to that obtained with RAP in monotherapy at a threefold higher dose (1 mg/kg/d). When the two drugs were given using an alternate day administration schedule (RAP at 0.6 mg/kg and 1,25(OH)2D3 at 2 micrograms/kg, each given on alternate days from day -3 to 19), near total protection was again obtained (13% paralysis, p < 0.001 versus control). These in vitro and in vivo data support the existence of synergistic interactions between 1,25(OH)2D3 and RAP. Considering the narrow therapeutic windows of both RAP and vitamin D-related compounds in autoimmune disease models, combinations of these drugs could find clinical application in reducing their individual therapeutically efficient doses to non-toxic levels. PMID- 9394787 TI - Embryonic expression of the mRNA for the rat homologue of the fusin/CXCR-4 HIV-1 co-receptor. AB - We have previously cloned a human receptor recently shown to be a cofactor for entry of T-tropic HIV-1 strains into CD4+ cells, now named fusin. Stromal derived factor-1 (SDF-1) is an endogenous ligand for fusin, also called CXCR-4. Here we show the distribution of fusin/CXCR-4 mRNA during ontogeny in the rat. The onset of mRNA expression is around embryonic day 9 and the mRNA expression is high in the thymus as well as proliferative areas of the brain during development. Our results suggest: (1) that fusin/CXCR-4 might have a dual role in both brain development and the immune system; (2) that SDF-1 has a role in brain development or that additional physiological ligands exist for this receptor; (3) co expression of CD4 and fusin/CXCR-4 may make fetuses susceptible to HIV infection during development. PMID- 9394788 TI - Compartmentalization of antigen specific cytokine responses to the central nervous system in CNS borreliosis: secretion of IFN-gamma predominates over IL-4 secretion in response to outer surface proteins of Lyme disease Borrelia spirochetes. AB - The neurological manifestations of Lyme disease have been proposed to be partly due to cytokine-mediated immunopathological mechanisms. In this study, the number of Borrelia-specific cells secreting interferon-gamma and interleukin-4 was determined in blood and cerebrospinal fluid from patients with CNS borreliosis (n = 23), other neurological diseases (n = 20), and in blood from healthy controls (n = 10), utilizing an ELISPOT-assay. Elevated specific secretion of IFN-gamma was found in CNS borreliosis, most pronounced in cerebrospinal fluid, whereas secretion of IL-4 was strikingly low. This may indicate that symptoms are due to side effects of the immune response, since IFN-gamma secretion in the absence of corresponding levels of IL-4 may be associated with tissue destruction. PMID- 9394789 TI - Dissociation of microglial activation and neuropathic pain behaviors following peripheral nerve injury in the rat. AB - Peripheral nerve injury commonly leads to neuropathic pain states fostered, in part, by neuroimmunologic events. We used two models of neuropathic pain (L5 spinal nerve cryoneurolysis (SPCN) and chronic constriction injury (CCI)) to assess the role of spinal glial activation responses in producing pain behaviors. Scoring of glial responses subjectively encompassed changes in cell morphology, cell density and intensity of immunoreactivity with specific activation markers (OX-42 and anti-glial fibrillary acidic protein (GFAP) for microglia and astrocytes, respectively). Glial responses were compared with tactile sensitivity (mechanical allodynia) at 1, 3 or 10 days following SPCN and with thermal hyperalgesia at 10 days in the CCI group. Neuropathic pain behaviors preceded and did not closely correlate with microglial responses in either model. Perineural application of bupivacaine prior to SPCN prevented spinal microglial responses but not pain behaviors. Spinal astrocytic responses to SPCN were early, robust and not altered by bupivacaine. The current findings support the use of bupivacaine as a tool to suppress microglial activation and challenge the putative role of microglia in initiating or potentiating pain behaviors which result from nerve injury. PMID- 9394790 TI - Thymocytes and cultured thymic epithelial cells express transcripts encoding alpha-3, alpha-5 and beta-4 subunits of neuronal nicotinic acetylcholine receptors: preferential transcription of the alpha-3 and beta-4 genes by immature CD4 + 8 + thymocytes. AB - Thymic tissues express transcripts encoding the alpha-3, alpha-5 and beta-4 subunits of nicotinic neuronal acetylcholine receptors (AcChRs) suggesting that neuronal AcChRs similar to those expressed in ganglia are expressed in the thymus. Transcription occurs in both isolated thymocytes and thymic epithelial cells. RT-PCR analyses of thymocyte subsets indicate that immature CD4 + 8 + thymocytes express higher levels of the alpha-3 and beta-4 transcripts than more mature thymocytes. Compared to freshly isolated thymocytes, peripheral blood lymphocytes do not express alpha-3 and beta-4 AcChR subunit transcripts. Cultured thymocytes rapidly down-regulate transcription of the alpha-3 and beta-4 AcChR subunit genes by a process that is not reversed by stimulation with phytohemagglutinin and IL-2. Thus our results indicate that there is transcriptional regulation of neuronal AcChR subunit genes during the process of thymocyte maturation and that factors within the thymic microenvironment influence expression of the alpha-3 and beta-4 AcChR subunit genes by developing T cells. PMID- 9394791 TI - Oligoclonal expansion of muscle infiltrating T cells in inclusion body myositis. AB - Inclusion body myositis (IBM) is the most common muscle disease affecting individuals over 50 years of age. An important feature of IBM is invasion of muscle fibers by T cells. The muscle infiltrating T cells show a restricted usage of variable (V) alpha/beta gene families. In this study we have investigated the clonality of T cells using two of the predominant V beta families i.e. V beta 3 and V beta 8 in three patients with IBM. The study was performed by reverse transcription and polymerase chain reaction (RT-PCR) analysis, followed by cloning and sequencing of the T cell receptor complementarity determining region 3. We found oligoclonal expansion of V beta 3 bearing muscle infiltrating T cells in two patients and of V beta 8 in one patient, supporting the concept that antigen stimulated T cells are important in the pathogenesis of IBM. Results of HLA typing indicated a genetic predisposition for the disease by the presence of DR3, DR52 and DQB1*0201/0202 in all three patients. PMID- 9394792 TI - Ia expression and antigen presentation by glia: strain and cell type-specific differences among rat astrocytes and microglia. AB - Astrocytes from experimental allergic encephalomyelitis (EAE)-susceptible Lewis rats expressed higher levels of Interferon-gamma-inducible Ia than astrocytes from EAE-resistant Brown Norway (BN) rats, whereas BN microglia expressed higher Ia than Lewis at both mRNA and protein levels. Lewis astrocytes induced proliferation of MBP-specific T cells selected on Lewis background as efficiently as Lewis thymocytes, whereas BN astrocytes were much less efficient in stimulating T cells selected in the presence of BN thymocytes. Microglia, irrespective of strain, induced only weak proliferative responses of these T cells despite the high expression of Ia. Antigen-stimulated T cells underwent apoptosis in the presence of microglia but not astrocytes. Thus, astrocyte mediated proliferation of MBP-specific T cells may contribute to the development of EAE, while microglia-induced T cell apoptosis may downregulate immunopathological processes in the brain. PMID- 9394793 TI - Paraneoplastic anti-Hu serum: studies on human tumor cell lines. AB - Patients with low titers of anti-Hu, the paraneoplastic encephalomyelitis/sensory neuronopathy (PEM/PSN) antibody, have a better tumor prognosis that those who do not harbor these antibodies. Accordingly, we examined the effects of serum from patients with anti-Hu antibodies on human tumor cell lines, in order to determine: (1) if the serum was toxic (growth inhibition or cytolysis) to tumor cells with or without complement, and (2) if anti-Hu antibodies contributed to tumor toxicity. The serum of 14 patients with anti-Hu associated PEM/PSN, 22 patients with small-cell lung cancer (SCLC) without anti-Hu antibodies, and 20 normal individuals were studied. Three cell lines (NT-2, BE(2)-C, and SH.SY5Y) that express Hu proteins, and one cell line (SAOS-2) that does not, were studied. We examined the effects of whole serum, IgG-depleted serum, and purified IgG in the presence or absence of complement. A higher percentage of anti-Hu sera were toxic (71%) compared with sera from anti-Hu negative SCLC patients (23%) (p < 0.0001). No correlation existed between the titer of anti-Hu antibodies and toxicity. The toxic effects were observed in all tumor cell lines including the cell line that does not express Hu antigens. Toxicity persisted in serum depleted of IgG. Purified anti-Hu IgG in the presence and absence of complement, was not toxic. Our findings indicate that anti-Hu serum is toxic for human tumor cell lines, but this toxicity does not appear to be mediated by anti-Hu antibodies. PMID- 9394794 TI - The restraint stress-induced reduction in lymphocyte cell number in lymphoid organs correlates with the suppression of in vivo antibody production. AB - In this study, we examined the effects of restraint stress on some immune parameters such as the in vivo antibody levels, cytokine production, and lymphocyte cell number in the spleen or mesenteric lymph node (MLN). BALB/c mice were thus injected intraperitoneally 2-times with OVA absorbed into alum on days 0 and 21. Before the first injection, the animals were either restrained for 12 h (stress group) or returned to their home cage (control group). Exposure to stress resulted in a reduction in the serum levels of anti-OVA IgE, IgG1, and IgG2a. In addition, stress also caused a decrease in the IL-4 and IFN-gamma levels in the spleen or mesenteric lymph node cell culture supernatants. Furthermore, exposure to stress resulted in a decrease in the splenic and mesenteric lymphocyte cell number when examined immediately after the cessation of stress. This decrease persisted for at least 12 h after the termination of stress and thereafter disappeared 24 h after stress. The stress-induced reductions in antibody and cytokine production occurred only when antigen was given either immediately or 6 h after stress, but not when antigen was given 24 h post stress. These results thus suggest that the restraint stress-induced change in lymphocyte cell number in the spleen or MLN closely correlates with the altered antibody and cytokine levels. PMID- 9394795 TI - A re-evaluation of the effects of X-linked immunodeficiency (xid) mutation on B cell differentiation and function in the mouse. AB - CBA/N (xid) mice have a point mutation in Bruton's tyrosine kinase (btk), which results in their failure to respond to T-independent type 2 (TI-2) antigens, and to several B cell mitogens [most notably anti-immunoglobulin (Ig)] in vitro. They have reduced numbers of peripheral (B2) B cells, which are regarded as being phenotypically and functionally immature. We show here that adult CBA/N mice in fact have two distinct B cell populations: some 60% of the cells are CD23+ HSAlo sIgDhi and hence resemble recirculating, follicular (RF) B cells from normal mice, except that they are sIgMhi. The remaining 40% of xid B cells are CD23- HSAhi sIgD-/lo and resemble immature transitional (TR) B cells. TR B cells from xid mice do not synthesize DNA when cultured with lipopolysaccharide (LPS), whereas those from normal mice do so. Only the RF cells from either xid or normal mice proliferate in response to ligation of CD40. In neonatal normal mice the emergence of mitogen responsiveness followed the chronological sequence LPS- >anti-CD40-->anti-Ig approximately anti-CD38. The same developmental sequence was seen with B cells from xid mice (for LPS and anti-CD40), but it occurred at a significantly slower tempo and this correlated with the later appearance of RF type cells. TR xid B cells express very low levels of bcl-2 and we conclude that these cells resemble very immature (bone marrow) B cells, rather than normal transitional cells. We, therefore, propose that the xid mutation imposes a multistage brake on B cell differentiation in the mouse. The available data suggest that btk is required for the positive selection of B cells throughout their differentiation in the periphery. This in turn implies that low level signaling via surface Ig is needed throughout this process in order for peripheral B cells to become functionally mature. PMID- 9394796 TI - A role for the VLA-4 integrin in the activation of human memory B cells. AB - It is generally recognized that activation through membrane effector molecules such as CD40 or the B cell receptor (BCR) is mandatory to allow B cells to proliferate and differentiate into antibody (Ab)-secreting cells in response to cytokines. We show here that purified tonsillar B cells can be stimulated directly by a cytokine combination to proliferate and secrete immunoglobulins when cultures are performed at high cell density. The contact-mediated activation of B cells in this experimental system is strongly inhibited both by anti-very late antigen (VLA)-4 monoclonal Ab and by a peptide containing the LDV sequence specifically recognized by the alpha 4 integrin binding site. These reagents also significantly suppressed the B cell responses elicited by engagement of the BCR or CD40. Our data reveal that memory B cells but not virgin or germinal center B cells are sensitive to the direct stimulatory effect of cytokines in high-density cultures. Finally, we found that the dual expression of the alpha and beta chains of VLA-4 is a distinctive feature of the memory B cell population. Collectively, our findings support the notion that VLA-4-dependent homotypic B cell interactions can mediate a co-stimulatory signal to human memory B cells and might participate in the B cell activation triggered through the BCR and CD40. PMID- 9394797 TI - In vitro cell death of activated lymphocytes in Omenn's syndrome. AB - Omenn's syndrome (OS) is characterized by erythrodermia, hepatosplenomegaly, lymphadenopathy, hypereosinophilia and elevated IgE levels associated with increased susceptibility to severe infections. Peripheral blood T cells, though usually present in normal number, show an activated phenotype (including an increased expression of CD95/Fas), a Th2 pattern of cytokine secretion and defective proliferative response to mitogens. In this report, we demonstrate that T cells from patients with OS undergo an excessive cell death in vitro resulting from two mechanisms. First, a substantial number of peripheral blood lymphocytes from OS patients die in unstimulated cultures (p = 0.009 vs. healthy controls). This spontaneous apoptosis is associated with reduced expression of bcl-2 gene product (p < 0.05) and can be prevented by addition of interleukin (IL)-2 (which also prevents down-modulation of bcl-2), while is independent from CD95 signaling. Second, lymphocytes from OS patients are highly susceptible to activation-induced cell death (AICD) induced with mitogens. This mechanism is largely independent from IL-2, while it can be significantly inhibited blocking CD95 with an IgG2b monoclonal antibody (mAb). The dependence of AICD from signals transduced via CD95 was confirmed showing that cross-linking CD95 with an IgM mAb induces a higher cell death in purified CD4+ CD45R0+ cells from OS patients than in controls (comparable for CD95 expression). Both mechanisms of cell death observed in this study result from lymphocyte hyperactivation occurring in vivo in these patients and may contribute to functional T cell defects of OS. PMID- 9394798 TI - Activation induces apoptosis in Herpesvirus saimiri-transformed T cells independent of CD95 (Fas, APO-1). AB - Signaling via the T cell receptor (TCR)/CD3 complex of pre-activated T cells induces apoptosis. Such an activation-induced cell death (AICD) is thought to play an important role in the regulation of cellular immune responses. In this study we analyzed pathways of AICD by using human T cells transformed by Herpesvirus saimiri. These growth-transformed T cells show the phenotype of activated mature T cells and continue to express a functionally intact TCR. We show that human H. saimiri-transformed T cell clones readily undergo cell death upon signaling via the TCR/CD3 complex or via phorbol 12-myristate 13-acetate (PMA) + ionomycin. The AICD in H. saimiri-transformed T cells was detectable a few hours after activation and it was not affected by the presence of interleukin (IL)-2 or by anti-CD4 cross-linking. However, AICD required tyrosine phosphorylation, since it could be blocked by herbimycin A. Cyclosporin A (CsA) did not block the development of AICD, but other consequences of activation in H. saimiri-transformed T cells like the production of interferon-gamma. Surprisingly, the development of AICD was not reduced by neutralizing antibodies to tumor necrosis factor (TNF)-alpha or blocking antibodies directed to CD95 (Fas, APO-1), although H. saimiri-transformed T cells were sensitive to CD95 ligation. To confirm that this form of AICD is really independent of CD95, we have established an H. saimiri-transformed T cell line from a patient with a homozygous deletion in the CD95 gene. This CD95-deficient T cell line was as sensitive to AICD as other CD95-expressing H. saimiri-transformed T cells. In conclusion, we describe here a type of AICD in H. saimiri-transformed T cells that is independent of CD95 and TNF-alpha, not sensitive to CsA, but requires tyrosine phosphorylation. This system should be useful for the investigation of CD95-independent forms of AICD. PMID- 9394799 TI - Rapamycin inhibits proteasome activator expression and proteasome activity. AB - Rapamycin (RAPA) is a potent immunosuppressive drug, and certain of its direct or indirect targets might be of vital importance to the regulation of an immune response. In this study, we used differential hybridization to search for human genes whose expression was sensitive to RAPA. Seven RAPA-sensitive genes were found and one of them encoded a protein with high homology to the alpha subunit of a proteasome activator (PA28 alpha). This gene was later found to code for the beta subunit of the proteasome activator (PA28 beta). Activated T and B cells had up-regulated PA28 beta expression at the mRNA level. Such up-regulation could be suppressed by RAPA, FK506, and cyclosporin A. RAPA and FK506 also repressed the up-regulated PA28 alpha messages in phytohemagglutinin (PHA)-stimulated T cells. At the protein level, RAPA inhibited PA28 alpha and PA28 beta in the activated T cells according to immunoblotting and confocal microscopy. Probably as a consequence, there was a fourfold increase of proteasome activities in the peripheral blood mononuclear cell lysate after the PHA activation. RAPA could inhibit the enhanced part of the proteasome activity. Considering the critical role played by the proteasome in degrading regulatory proteins, our data suggest that the proteasome activator is a relevant and important downstream target of rapamycin, and that the immune response could be modulated through the activity of the proteasome. PMID- 9394800 TI - Comparison of the effects of interleukin-1 alpha, interleukin-1 beta and interferon-gamma-inducing factor on the production of interferon-gamma by natural killer. AB - Interferon-gamma inducing factor (IGIF) is a recently identified cytokine which stimulates the production of interferon-gamma (IFN-gamma) by T cells and enhances natural killer (NK) cell cytolytic activity. Protein fold recognition, structure prediction and comparative modeling have revealed that IGIF is a member of the interleukin (IL)-1 cytokine family and has prompted the designation IL-1 gamma. Here we report functional similarities between members of the IL-1 family by comparing the effects of IL-1 alpha, IL-1 beta and IGIF on NK cell production of IFN-gamma. All three IL-1 types enhanced NK cell production of IFN-gamma when induced by IL-2 or IL-12, although at high concentrations (> 10 ng/ml), IGIF was five- to tenfold more potent than IL-1 alpha or IL-1 beta. This effect correlated with enhanced levels of mRNA for IFN-gamma when NK cells were stimulated with IGIF plus IL-12. In contrast to IL-12 and IL-2, the ability of IGIF to stimulate NK cell production of IFN-gamma was not increased by IL-1 alpha or IL-1 beta. The ability of IGIF to enhance IFN-gamma production was independent of the type I and type II IL-1 receptors or the IL-1R accessory protein. Together, these results identify IGIF as a potent stimulator of NK cell production of IFN-gamma and demonstrate that the effect of IGIF on NK cell production of IFN-gamma is similar to that of IL-1 alpha and IL-1 beta but distinct from that of IL-12. PMID- 9394801 TI - Rho prevents apoptosis through Bcl-2 expression: implications for interleukin-2 receptor signal transduction. AB - Here we describe a Rho-mediated apoptosis suppression pathway driven by Bcl-2 expression in the interleukin (IL)-4- or IL-2-dependent murine T cell line TS1 alpha beta. IL-2, but not IL-4, induces Bcl-2 expression through RhoA activation which is inhibited by the specific Rho family inhibitor, Clostridium difficile Toxin B, as well as by a dominant negative RhoA mutant. Using transient transfections of RhoA mutants tagged with the vesicular stomatitis virus glycoprotein, we show that a constitutively active RhoA mutant induces Bcl-2 expression and prevents apoptosis upon IL-4 withdrawal. Finally, we have identified the signaling pathway involved together with RhoA in Bcl-2 induction and show compelling evidence for the implication of phosphatidylinositol 3 kinase and protein kinase C. PMID- 9394802 TI - Involvement of nuclear factor-kappa B in platelet-activating factor-mediated tumor necrosis factor-alpha expression. AB - Tumor necrosis factor (TNF)-alpha and platelet-activating factor (PAF) are important mediators of inflammatory reactions, and their release is controlled by a positive feedback network. However, the regulatory mechanisms underlying the interaction of these two molecules are unknown. Within 10 min of the injection of lipopolysaccharide (LPS) into C57BL/6 mice, effects inducible by PAF such as anaphylactic shock-like symptoms, disseminated intravascular coagulation, and hemorrhage in renal medullae were observed, and all these pathological changes were prevented by the PAF antagonist, BN 50739. The plasma level of PAF after LPS injection reached a peak at 5 min. TNF-alpha gene expression was evident 20 min after LPS injection and was maximal at 40 min, and the level of serum TNF-alpha reached a peak at 1 h. Pretreatment with BN 50739 inhibited LPS-induced TNF-alpha gene expression and protein synthesis in a dose-dependent manner. Injection of PAF or treatment of the macrophage cell line, J774A.1, with PAF activated the transcription factor, nuclear factor (NF)-kappa B, which is essential for inducible TNF-alpha transcription. The activation of NF-kappa B by PAF preceded the LPS-mediated TNF-alpha gene expression. Pretreatment with BN 50739 inhibited LPS-induced mobilization of NF-kappa B in a dose-dependent manner in vivo as well as in vitro. These data suggest that PAF, which is released immediately or shortly after LPS injection, induces the expression of TNF-alpha through the activation of NF-kappa B. PMID- 9394803 TI - Extracellular HIV-1 Tat protein activates phosphatidylinositol 3- and Akt/PKB kinases in CD4+ T lymphoblastoid Jurkat cells. AB - The biological basis for the pleiotropic activity of extracellular human immunodeficiency virus (HIV)-1 Tat protein on lymphoid T cell survival is not well understood. We have here demonstrated that the addition in culture of 0.1-10 nM Tat protein to 36-h serum-starved lymphoblastoid Jurkat T cells rapidly stimulates the catalytic activity of phosphatidylinositol 3-kinase (PI 3-K). The peak of activation was observed 30 min after Tat addition. Extracellular Tat also stimulated the catalytic activity of the Akt/PKB kinase, a major target of PI 3-K lipid products. Pretreatment of serum-starved Jurkat cells with 100 nM wortmannin (WT) or 10 microM LY294002, two unrelated pharmacological inhibitors of PI 3-K, markedly suppressed the catalytic activity of both PI 3-K and Akt/PKB in Jurkat cells. Moreover, at low concentrations (0.1-1 nM), extracellular Tat showed a small but reproducible protection of Jurkat cells from apoptosis induced by serum deprivation (p < 0.05), while the combination of Tat plus 100 nM WT significantly (p < 0.05) increased the percentage of apoptosis with respect to cells left untreated or treated with Tat alone. Taken together, these data suggest that the anti-apoptotic activity of low concentrations of Tat protein on Jurkat cells is mediated by a PI 3-kinase/Akt pathway. PMID- 9394804 TI - Control of self-reactive cytotoxic T lymphocytes expressing gamma delta T cell receptors by natural killer inhibitory receptors. AB - The majority of peripheral blood gamma delta T cells in human adults expresses T cell receptors (TCR) with identical V regions (V gamma 9 and V delta 2). These V gamma 9 V delta 2 T cells recognize the major histocompatibility complex (MHC) class I-deficient B cell line Daudi and broadly distributed nonpeptidic antigens present in bacteria and parasites. Here we show that unlike alpha beta or V gamma 9- gamma delta T cells, the majority of V gamma 9V delta 2 T cells harbor natural killer inhibitory receptors (KIR) (mainly CD94/NKG2A heterodimers), which are known to deliver inhibitory signals upon interaction with MHC class I molecules. Within V gamma 9V delta 2 T cells, KIR were mainly expressed by clones exhibiting a strong lytic activity against Daudi cells. In stark contrast, almost all V gamma 9V delta 2 T cell clones devoid of killing activity were KIR-, thus suggesting a coordinate acquisition of KIR and cytotoxic activity within V gamma 9V delta 2 T cells. In functional terms, KIR inhibited lysis of MHC class I positive tumor B cell lines by V gamma 9V delta 2 cytotoxic T lymphocytes (CTL) and raised their threshold of activation by microbial antigens presented by MHC class I-positive cells. Furthermore, masking KIR or MHC class I molecules revealed a TCR-dependent recognition by V gamma 9V delta 2 CTL of ligands expressed by activated T lymphocytes, including the effector cells themselves. Taken together, these results suggest a general implication of V gamma 9V delta 2 T cells in immune response regulation and a central role of KIR in the control of self-reactive gamma delta CTL. PMID- 9394805 TI - Requirement of a second signal via protein kinase C or protein kinase A for maximal expression of CD40 ligand. Involvement of transcriptional and posttranscriptional mechanisms. AB - High levels of CD40 ligand (CD40L) protein expression are induced on native T cells by increasing the intracellular Ca2+ concentration. In the present study we have shown that ionomycin induces CD40L gene transcription leading to mRNA accumulation which translates to high levels of protein expression. Conversely, agents which increase the intracellular levels of cyclic AMP (cAMP), such as prostaglandin E2 (PGE2) or dibutyryl cyclic AMP (dbcAMP), were unable to induce CD40L expression on T lymphocytes. Cell activation by phorbol 12-myristate 13 acetate (PMA) treatment had a slight effect on increasing CD40L mRNA and protein levels. However, PMA and dbcAMP synergized with ionomycin to significantly increase and to prolong the CD40L expression. Nuclear run-on assays revealed that PMA, but not dbcAMP, increased threefold the CD40L gene transcription rate induced by ionomycin. This effect was independent of de novo protein synthesis. In addition, at a posttranscriptional level, both reagents synergized with the Ca2+ ionophore to prolong the CD40L mRNA half-life by a mechanism which was also independent of de novo protein synthesis. Moreover, when transcription was blocked with actinomycin D, an increment of the CD40L transcript levels induced by PMA or dbcAMP on ionomycin-treated cells was observed in the presence of cycloheximide. This probably means that newly synthesized protein may contribute to the CD40L mRNA destabilization. In summary, these data show that PMA and dbcAMP synergized with ionomycin to increase the CD40L mRNA and protein levels. The up-regulatory effect of PMA was accomplished at a transcriptional and posttranscriptional level, whereas dbcAMP exerted its synergistic effect exclusively at a posttranscriptional level. PMID- 9394806 TI - Induction of heat shock protein 72 synthesis by endogenous tumor necrosis factor via enhancement of the heat shock element-binding activity of heat shock factor 1. AB - Endogenous tumor necrosis factor (enTNF) acts as a resistance factor against cytotoxicity caused by heat by inducing manganous superoxide dismutase (MnSOD), thereby scavenging reactive oxygen free radicals. On the other hand, it is also well known that heat shock proteins (HSP) which are induced by heat stress behave as cytoprotective factor against this stress. However, the relationship of these two resistance factors is not elucidated yet. In the present study, we therefore proposed the possibility that enTNF enhances HSP72 expression. Heat-sensitive L-M (mouse tumorigenic fibroblast) cells, which normally do not express enTNF, were transfected with a nonsecretory-type human TNF-alpha expression vector to produce enTNF. Stable transfectants showed resistance to heat treatment and an increase of HSP72 expression. Conversely, when HeLa (human uterine cervical cancer) cells, which normally produce an appreciable amount of enTNF, were transfected with an antisense TNF-alpha mRNA expression vector to inhibit enTNF synthesis, their heat sensitivity was enhanced and HSP72 expression was reduced by half. Although enTNF caused no difference in the level of heat shock factor (HSF) 1 in these cells, enTNF expression correlated well with the binding activity of HSF-1 to a 32P labeled synthetic oligonucleotide containing the human heat shock element (HSE). These results indicate that enTNF participates not only in intrinsic resistance against heat via induction of MnSOD but also via enhancement of the HSE-binding activity of HSF 1 followed by augmentation of HSP72 expression. PMID- 9394807 TI - Cloning of NKG2-F, a new member of the NKG2 family of human natural killer cell receptor genes. AB - The NKG2 family of genes encodes at least four different type II transmembrane molecules (NKG2-A, NKG2-B, NKG2-C and NKG2-E) which contain a C-lectin domain. These proteins have been shown to be covalently associated with CD94, another C type lectin member. The heterodimers are involved in natural killer cell-mediated recognition of different HLA-allotypes. Here we describe the cloning of a new NKG2-related gene, termed NKG2-F, localized 25 kb from NKG2-A as well as its relationship with the previously described NKG2-D cDNA. Despite the similarities with the other NKG2 genes, NKG2-F encodes a putative protein which does not contain any lectin domain. However, a conserved 24-amino acid sequence, present in all members of the NKG2 family, suggests that NKG2-F is also able to form heterodimers with CD94. PMID- 9394808 TI - The central nervous system environment controls effector CD4+ T cell cytokine profile in experimental allergic encephalomyelitis. AB - In experimental allergic encephalomyelitis (EAE), CD4+ T cells infiltrate the central nervous system (CNS). We derived CD4+ T cell lines from SJL/J mice that were specific for encephalitogenic myelin basic protein (MBP) peptides and produced both Th1 and Th2 cytokines. These lines transferred EAE to naive mice. Peptide-specific cells re-isolated from the CNS only produced Th1 cytokines, whereas T cells in the lymph nodes produced both Th1 and Th2 cytokines. Mononuclear cells isolated from the CNS, the majority of which were microglia, presented antigen to and stimulated MBP-specific T cell lines in vitro. Although CNS antigen-presenting cells (APC) supported increased production of interferon (IFN)-gamma mRNA by these T cells, there was no increase in the interleukin (IL) 4 signal, whereas splenic APC induced increases in both IFN-gamma and IL-4. mRNA for IL-12 (p40 subunit) was up-regulated in both infiltrating macrophages and resident microglia from mice with EAE. We have thus shown that a Th1 cytokine bias within the CNS can be induced by CNS APC, and that IL-12 is up-regulated in microglial cells within the CNS of mice with EAE. Microglia may therefore control Th1 cytokine responses within the CNS. PMID- 9394809 TI - Influence of complement on the allospecific antibody response to a primary vascularized organ graft. AB - The induction of antibody responses against T cell-dependent antigens has been reported to be influenced by complement. We therefore asked if the primary induction of alloantibodies against transplantation antigens, an important determinant of transplant outcome, is complement sensitive and whether this has functional implications. We transplanted rat kidney allografts into fully major histocompatibility complex-mismatched recipients, in which complement activation was inhibited by daily injection of soluble recombinant human complement receptor type 1 (sCR1). Control allograft recipients were injected with saline. Animals in the control group showed a marked antibody response against donor-specific antigens and an increase in the proportion of activated B and T splenocytes by day 5 after transplantation. Complement-inhibited rats showed a reduced level of antibody binding on target cells sharing the same histocompatibility antigens as the donor strain (p < 0.001), and a reduced level of activated splenic B (p < 0.01) and T (p < 0.01) cells. In a functional assay, the plasma of complement inhibited rats showed reduced cytotoxic activity against donor-specific cells, and their grafts contained less bound antibody than controls. Analysis beyond 6 days was obscured due to the development of antibodies against sCR1. We conclude that complement activation facilitates the induction of the alloantibody response. Sparing of vascular injury and prolongation of graft survival, previously reported in complement-inhibited rats (Pratt J. R. et al., Am. J. Path. 1996, 149: 2055), could therefore be due to down-regulation of the B cell response as well as reduced complement-dependent cytotoxicity. Inhibition of complement may provide an ancillary approach to the prevention of allospecific antibody formation and the prolongation of allograft survival in primary kidney grafting. PMID- 9394810 TI - Analysis of susceptibility of NOD mice to spontaneous and experimentally induced thyroiditis. AB - Beside diabetes, non-obese diabetic (NOD) mice develop sporadic lymphoid infiltration of the thyroid gland, mimicking Hashimoto's thyroiditis. We have examined the prevalence of those manifestations in NOD mice, the influence of the major histocompatibility complex (MHC) and the association with autoantibodies. The incidence at 1 year is of 14.3% in wild-type NOD mice versus 19.6% in congenic NOD.H2k mice. The moderate, but statistically significant difference, based on the analysis of 161 NOD and 169 NOD.H2k mice, suggests that MHC genes partially control spontaneous NOD thyroiditis. Autoantibodies against thyroglobulin (Tg) are mouse specific and their presence correlates closely with thyroiditis. The strong correlation between cellular and humoral anomalies therefore resembles Hashimoto's thyroiditis. NOD and NOD.H2k mice actively immunized against Tg develop severe chronic lesions with epithelium necrosis and interstitial tissue fibrosis. Most interestingly, those lesions do not regress spontaneously as in CBA/J mice. Paradoxically, the response to Tg of lymph node cells from NOD mice is weaker both in proliferation and cytokine production. The defect is most evident for interferon-gamma-producing T cells and is reflected in the marked deficit in IgG2a antibodies. Thus a moderate anti-Tg response seems to favor chronicity of thyroiditis. In conclusion, NOD and NOD.H2k mice offer a unique opportunity of analyzing the factors leading to immune chronicity in a genetic context which promotes autoimmune endocrinopathies. PMID- 9394811 TI - Nitric oxide and the immunomodulation of experimental allergic encephalomyelitis. AB - Previous studies examining the effect of nitric oxide synthase (NOS) inhibition on the course of experimental allergic encephalomyelitis (EAE) have yielded conflicting results. This may relate to the use of nonspecific inhibitors and to differences between active and adoptive EAE. We examined the effect of treatment with L-N-(1-iminoethyl)lysine (L-NIL), a selective inhibitor of the cytokine inducible isoform of NOS, on the clinical course of active and adoptive EAE in Lewis rats. We find that while L-NIL treatment of recipients is protective in adoptive EAE, treatment of active EAE with L-NIL leads to a marked accentuation of disease expression. In L-NIL-treated animals treated with myelin basic protein/complete Freund's adjuvant (MBP/CFA), disease onset is accelerated and clinical symptoms are more severe. Accentuation of integrated disease scores is seen even if L-NIL treatment is started 5 days following immunization. The histological findings in involved spinal cords from L-NIL-treated animals with active EAE are similar to those from untreated animals with similar clinical scores. L-NIL treatment of MBP/CFA-immunized animals does not prevent recovery from clinical symptoms, nor does it allow for reinduction of disease in animals previously immunized with MBP/CFA. Treatment of F344 rats, a strain which is relatively nonsusceptible for EAE, with L-NIL results in consistent evidence of EAE following immunization with MBP/CFA. These findings, together with our previous work on interstitial nephritis, support a role for endogenously generated NO in immunoregulation of T cell responses following immunization with antigen in CFA, and suggest that inducibility of NOS expression may be an important susceptibility factor for autoimmunity. PMID- 9394812 TI - In vivo evidence for a functional role of both tumor necrosis factor (TNF) receptors and transmembrane TNF in experimental hepatitis. AB - The significance of tumor necrosis factor receptor 1 (TNFR1) for TNF function in vivo is well documented, whereas the role of TNFR2 so far remains obscure. In a model of concanavalin A (Con A)-induced, CD4+ T cell-dependent experimental hepatitis in mice, in which TNF is a central mediator of apoptotic and necrotic liver damage, we now provide evidence for an essential in vivo function of TNFR2 in this pathophysiological process. We demonstrate that a cooperation of TNFR1 and TNFR2 is required for hepatotoxicity as mice deficient of either receptor were resistant against Con A. A significant role of TNFR2 for Con A-induced hepatitis is also shown by the enhanced sensitivity of transgenic mice overexpressing the human TNFR2. The ligand for cytotoxic signaling via both TNF receptors is the precursor of soluble TNF, i.e. transmembrane TNF. Indeed, transmembrane TNF is sufficient to mediate hepatic damage, as transgenic mice deficient in wild-type soluble TNF but expressing a mutated nonsecretable form of TNF developed inflammatory liver disease. PMID- 9394813 TI - Expression of the Ly49A gene in murine natural killer cell clones is predominantly but not exclusively mono-allelic. AB - The Ly49 family of natural killer (NK) cell receptors are major histocompatibility complex (MHC) class I-specific inhibitory receptors that are distributed to overlapping NK cell subsets. Extending earlier studies of polyclonal NK cell populations, we have employed an analysis of short-term NK cell clones from Ly49A heterozygous mice, to demonstrate that the Ly49A gene is usually expressed from one or the other allele in each Ly49A+ cell. However, we also detected a small percentage of clones that expressed both Ly49A alleles. The possibility that the colonies exhibiting bi-allelic Ly49A gene expression had been inoculated with more than one cell was ruled out by parallel analysis of clones isolated from mixtures of NK cells from Ly49A homozygous mice. The frequency of bi-allelic Ly49A+ clones suggested that the two Ly49A alleles in an NK cell are chosen for expression independently. These data are consistent with the proposal that mono-allelic Ly49A gene expression may arise as a consequence of a stochastic Ly49 gene activation mechanism. Analysis of Ly49A+ clones from MHC-different mice demonstrated that class I-deficient mice harbored a greater number of bi-allelic Ly49A+ cells than did H-2d mice, which express a Ly49A ligand. Although the numbers were insufficiently large for a clear assignment, H 2b mice may harbor an intermediate number of bi-allelic Ly49A+ NK cells. The effects of MHC expression on the prevalence of bi-allelic Ly49A+ cells suggest that an MHC-dependent education process modifies the Ly49 repertoire. PMID- 9394814 TI - Interleukin-15 preferentially promotes the growth of intestinal intraepithelial lymphocytes bearing gamma delta T cell receptor in mice. AB - Several cytokines including stem cell factor (SCF) and interleukin (IL)-7 are known to be required for development of gamma delta T cell receptor (TCR) intestinal intraepithelial lymphocytes (i-IEL) in mice. We show here the effects of IL-15 on the proliferation and maintenance of murine gamma delta i-IEL in vitro. gamma delta i-IEL constitutively expressed a high level of IL-15 receptor alpha mRNA and proliferated in response to IL-15 more vigorously than alpha beta i-IEL. V gamma/delta repertoire analysis revealed that IL-15, like IL-2, induced polyclonal expansion of gamma delta i-IEL, whereas gamma delta i-IEL responding to IL-7 showed a V gamma/delta repertoire skewed towards V gamma 1/V delta 4, V delta 5. IL-15 efficiently prevented gamma delta i-IEL from apoptosis induced by growth factor deprivation. This rescue was accompanied by up-regulation of Bcl-2 expression. These results suggest that IL-15 plays important roles in proliferation and maintenance of gamma delta i-IEL. PMID- 9394815 TI - The CC chemokine antagonist Met-RANTES inhibits eosinophil effector functions through the chemokine receptors CCR1 and CCR3. AB - Eosinophils are predominant effector cells not only in allergic diseases but also in connective tissue diseases. The recruitment of eosinophils to the site of inflammation and release of reactive oxygen species leading to tissue damage and propagation of the inflammatory response are mediated by chemokines. Thus, agents that would be able to inhibit or antagonize chemokine-induced eosinophil activation are interesting as therapeutical agents. We describe the effect of a chemokine receptor antagonist, Met-RANTES, on human eosinophil effector functions in response to RANTES, monocyte chemoattractant protein (MCP)-3 and eotaxin. Met RANTES was able to inhibit dose-dependently [Ca2+]i transients in eosinophils following stimulation with RANTES, MCP-3 and eotaxin. Whereas maximal and half maximal inhibitory effect of Met-RANTES following stimulation with RANTES and MCP 3 were observed at 2 micrograms/ml and 1 microgram/ml, respectively, maximal and half-maximal inhibitory effects of Met-RANTES in response to eotaxin were detected at 10 micrograms/ml and 3 micrograms/ml. Moreover, eotaxin-induced [Ca2+]i transients were only half reduced at a Met-RANTES concentration at which RANTES and MCP-3 were completely blocked. Besides its effect on [Ca2+]i transients, Met-RANTES dose-dependently inhibited actin polymerization in eosinophils following chemokine stimulation. Whereas Met-RANTES totally inhibited RANTES- and MCP-3-induced actin polymerization at 5 micrograms/ml, the eotaxin induced response was only reduced by 50%. However, Met-RANTES inhibited dose dependently the release of reactive oxygen species in response to RANTES, MCP-3 and eotaxin. Again, eotaxin-induced release of reactive oxygen species, however, was only half reduced at a Met-RANTES concentration (10 micrograms/ml) at which RANTES and MCP-3 were completely blocked. The results of this study show that (1) Met-RANTES is an effective and powerful antagonist of effector functions of human eosinophils following stimulation with RANTES, MCP-3 and eotaxin; (2) Met-RANTES seems to be able to antagonize the response of eosinophils through chemokine receptor 1 (CCR1) preferentially to CCR3; (3) Met-RANTES antagonizes eosinophil but not neutrophil effector functions and might be therefore of interest for a new therapeutical approach to prevent the invasion and destructive power of eosinophils in diseases that are accompanied by eosinophil infiltration such as allergic asthma and connective tissue diseases. PMID- 9394816 TI - Expression of epsilon germ-line gene transcripts and mRNA for the epsilon heavy chain of IgE in nasal B cells and the effects of topical corticosteroid. AB - We have studied the expression of the gene encoding the epsilon heavy chain of IgE in nasal B cells of hayfever patients. We developed probes to detect transcripts of the epsilon germ-line gene and the rearranged gene by in situ hybridization of biopsy sections from the nasal mucosa. We compared tissue from hayfever patients out of season with that of normal controls, and also of hayfever patients treated with topical corticosteroid (fluticasone propionate) or placebo for 6 weeks and then challenged with antigen. epsilon chain mRNA was expressed in an unexpectedly high proportion of nasal B cells of both hayfever patients and normal subjects. However, although similar numbers of B cells were found in both groups, the proportion of cells that express epsilon chain mRNA was several times higher in the hayfever patients. No transcripts of the epsilon germ line gene were detected in either group before allergen challenge. When hayfever patients were administered antigen locally, early (10-30 min) and late (1-24 h) symptoms ensued. After 24 h, coincident with an increase in the number of cells expressing mRNA for IL-4 in the tissue, epsilon germ-line gene transcripts appeared in the nasal B cells. The induction by allergen of IL-4 mRNA and epsilon germ-line gene transcripts was suppressed by fluticasone propionate treatment. Our results suggest that local IgE synthesis and cytokine regulation of heavy chain switching to IgE occur in the nasal mucosa. PMID- 9394817 TI - Nitric oxide pathway is induced by Fc epsilon RI and up-regulated by stem cell factor in mouse mast cells. AB - Murine stem cell factor (SCF) induces the differentiation of mucosal mast cells (MMC) into connective tissue mast cells (CTMC) and potentiates mediator release induced by aggregation of high-affinity IgE receptors (Fc epsilon RI). In the present work, we investigated the effect of Fc epsilon RI aggregation on nitric oxide (NO) pathway induction in the different subsets of mast cells, as well as the contribution of SCF in this induction. Inducible NO synthase (iNOs) expression was not evidenced in non-stimulated MMC obtained by culture of hematopoietic progenitors in the presence of interleukin-3, whereas IgE-antigen stimulated MMC expressed iNOs mRNA and protein and synthesized nitrites. Long term treatment of MMC with SCF, allowing them to differentiate into CTMC, induced iNOs expression in non-stimulated cells and up-regulated iNOs expression and generation of NO derivatives induced by IgE-antigen stimulation. Thus, NO derivatives generated by mast cells could participate in inflammatory reactions during allergic stimulation. PMID- 9394818 TI - The roles of complement receptors type 1 (CR1, CD35) and type 3 (CR3, CD11b/CD18) in the regulation of the immune complex-elicited respiratory burst of polymorphonuclear leukocytes in whole blood. AB - The binding of immune complexes (IC) to polymorphonuclear leukocytes (PMN) and the consequent respiratory burst (RB) were investigated in whole blood cell preparations suspended in 75% human serum, using flow cytometry. Blockade of the complement receptor (CR)1 receptor sites for C3b on whole blood cells using the monoclonal antibody (mAb) 3D9 resulted in a 1.9-fold increase in the IC-elicited PMN RB after 5 min of incubation, rising to 3.1-fold after 40 min. This enhancement was not due to increased IC deposition on PMN. Blockade of CR3 abrogated the mAb 3D9-induced rise in RB activity and inhibited the IC binding to PMN in a whole blood cell preparation, with or without mAb 3D9, by approximately 40% from 15-40 min while reducing their RB over 40 min to approximately one third. Blockade of CR1 on either erythrocytes (E) or leukocytes, before mixing the populations, revealed that the potentiation of the RB by mAb 3D9 was associated with abrogation of E-CR1 function, whereas blockade of leukocyte-CR1 had a diminishing effect. Exposure to IC at high concentrations induced release of both specific and azurophilic granule contents from PMN. The latter was CR3 dependent in that blockade of the receptor inhibited the lactoferrin release by one third during 40 min of incubation. In conclusion, CR3 plays a significant role in the IC-mediated generation of an RB and release of specific granules by PMN, while CR1 on whole blood cells, primarily E CR1, restricts the IC-elicited RB in PMN. We propose that CR1 in whole blood promotes the degradation of IC bound iC3b to C3dg, thereby rendering the IC inaccessible for binding to CR3. PMID- 9394819 TI - Structural analysis of human gamma 3 intervening regions and switch regions: implication for the low frequency of switching in IgG3-deficient patients. AB - High and low serum concentrations of IgG3 are associated with the human G3m(b) and G3m(g) allotypes, respectively. In the present study, we analyzed the structure of the S gamma 3 and I gamma 3, the switch frequency, switch breakpoints and the levels and initiation sites of I gamma 3 transcripts both in normal blood donors expressing (b) or (g) allotypes as well as IgG3-deficient (D) patients. A low switch frequency to gamma 3 was found in the (g) allotype IgG3D patients which may be caused in part by the allotype-associated mutations in the S gamma 3 region and in part by additional individual mutations observed in the A (SNAP) and B (SNIP/ NF-kappa B) sites in the S gamma 3 repeat region. A higher I gamma 3 germ-line (GL) transcriptional rate was seen in cells from the IgG3D patient, suggesting that low levels of GL I gamma 3 transcripts are not a major contributing factor to the defect. However, individual mutations in the I gamma 3 region and differential splicing of GL I gamma 3 transcripts were found which may affect the switching process. PMID- 9394820 TI - Relapsing and remitting experimental allergic encephalomyelitis: a focused response to the encephalitogenic peptide rather than epitope spread. AB - The progression of experimental allergic encephalomyelitis (EAE) in certain mouse strains has been reported to involve a broadening of the response to myelin antigens, apparently resulting from priming to endogenous determinants of the myelin sheath. The phenomenon has been termed determinant spread. Interest in this effect has centered on the mechanism it offers to explain the progressive, relapsing and remitting course of EAE and indeed of multiple sclerosis. We have conducted a systematic, longitudinal study in SJL mice to look for determinant spread during relapsing and remitting EAE, correlating epitope recognition and cytokine production with disease severity. Disease was induced using three of the four encephalitogenic proteolipid protein or myelin basic protein epitopes, and responses to each of four epitopes recognized by SJL T cells were tracked through acute disease, remission and relapse. The responses of lymph node cells, splenocytes and central nervous system (CNS)-infiltrating T cells were analyzed. While marginal, transient responses to secondary epitopes were detectable in splenocytes, CNS-infiltrating cells showed a dominant response to the original disease-inducing epitope without evidence of a shift to other determinants during relapse. Disease relapse was correlated with an increase in CNS-infiltrating cells and a high proliferative and interferon (IFN)-gamma response to the disease inducing peptide. During remission, there was a decrease in numbers of cells infiltrating the CNS. These cells were down-regulated, showing low if any response to the myelin peptides tested as measured by proliferation, production of IFN-gamma or production of IL-4. Our findings argue strongly against a causal role for determinant spread in disease relapse as observed in these models of EAE. PMID- 9394821 TI - In vivo rolling and endothelial selectin binding of mononuclear leukocytes is distinct from that of polymorphonuclear cells. AB - In inflammation, rolling of leukocytes along the microvascular endothelium is a precondition for subsequent integrin-mediated firm adhesion and extravasation. Rolling characteristics of polymorphonuclear leukocytes (PMNL) and mononuclear leukocytes (MNL) in small venules (15-25 microns) of the rat mesentery were studied by intravital fluorescence microscopy under basal conditions and after intravenous treatment with an anti-rat neutrophil serum (ANS). The baseline rolling fraction of the venular total leukocyte flux was 36 +/- 15% (mean +/- SD). The PMNL fraction of the systemic leukocyte count was 27 +/- 9%. Treatment with ANS resulted in total depletion of circulating PMNL and reduced the leukocyte rolling fraction to 12 +/- 5%, in this situation represented only by MNL. In rats treated intraperitoneally with interleukin (IL)-1 beta for 4 h, the leukocyte rolling fraction was 53 +/- 13% and was reduced to 33 +/- 11% after ANS treatment. These data indicated that most, if not all, circulating PMNL rolled along the venular endothelial lining in the rat mesentery prepared for intravital microscopy, whereas MNL rolling was minor (approximately 10%) under the same basal condition. In cytokine-activated tissue, on the other hand, the number of rolling MNL was greatly increased. While PMNL rolling is known to be entirely selectin dependent, the increased MNL rolling after IL-1 stimulation was likely mediated by alpha 4 integrins, inasmuch as the rolling fraction of isolated peripheral blood lymphocytes injected into the microcirculation of the cytokine stimulated mesentery was reduced from 31 +/- 14% to 6 +/- 2% by pretreatment of the cells with a monoclonal antibody against the rat integrin alpha 4 chain. In accordance with the in vivo rolling characteristics of the two cell populations, binding of soluble P- or E-selectin (selectin/IgG chimeras) was less intense for blood lymphocytes than for granulocytes, as determined by flow cytometric analyses of rat and human leukocytes. Taken together, our findings in vivo indicate that the adhesive interactions responsible for rolling of PMNL and MNL, respectively, are distinct in terms of receptor occupancy, and may help explain the temporal selectivity in recruitment of different leukocyte subpopulations in inflammatory or immune reactions. PMID- 9394822 TI - T helper cell priming of mice to Borrelia burgdorferi OspA leads to induction of protective antibodies following experimental but not tick-borne infection. AB - Antibodies to the outer surface lipoprotein A (OspA) of Borrelia burgdorferi confer protection to SCID mice against subsequent tick-borne or experimental infection. However, OspA-specific antibodies are hardly detectable in naturally infected humans, dogs, hamsters and mice. This is most probably due to limited expression of OspA on spirochetes transmitted from the vector to the host. Here we have tested whether T cell priming of mice would lead to the induction of protective OspA-specific antibodies upon infection. It is shown that AKR/N mice, previously immunized with either a single T helper cell peptide of OspA, or a mixture of 27 peptides spanning the entire molecule, develop OspA-specific IgM or IgG antibodies, including those to a prominent protective B cell epitope of OspA. LA-2, within 7 days of infection with low doses (10(3)) of culture-derived spirochetes. In marked contrast, the same groups of pre-sensitized mice failed to generate any detectable OspA-specific antibodies after tick-borne infection for more than 40 days after infection. All mice, irrespective of their state of T cell immunity to OspA or the mode of infection, produced similar levels of OspC specific IgM and IgG antibodies as early as day 14 after infection. None of the mice previously immunized with OspA peptides were protected against experimental infection, in spite of the appearance of protective antibodies. It is clear from these data that, in contrast to culture-derived spirochetes, the naturally transmitted pathogen fails to express OspA within the mammalian host at levels sufficient for induction of B cell responses, even in the presence of pre activated T helper cells. Together with the fact that OspA-specific antibodies are mainly operative by eliminating spirochetes from the vector during infestation, the data suggest that OspA-vaccination for T helper cell immunity alone is not sufficient to prevent Lyme disease. PMID- 9394823 TI - Alpha beta lineage-committed thymocytes can be rescued by the gamma delta T cell receptor (TCR) in the absence of TCR beta chain. AB - Commitment of the alpha beta and gamma delta T cell lineages within the thymus has been studied in T cell receptor (TCR)-transgenic and TCR mutant murine strains. TCR gamma delta-transgenic or TCR beta knockout mice, both of which are unable to generate TCR alpha beta-positive T cells, develop phenotypically alpha beta-like thymocytes in significant proportions. We provide evidence that in the absence of functional TCR beta protein, the gamma delta TCR can promote the development of alpha beta-like thymocytes, which, however, do not expand significantly and do not mature into gamma delta T cells. These results show that commitment to the alpha beta lineage can be determined independently of the isotype of the TCR, and suggest that alpha beta versus gamma delta T cell lineage commitment is principally regulated by mechanisms distinct from TCR-mediated selection. To accommodate our data and those reported previously on the effect of TCR gamma and delta gene rearrangements on alpha beta T cell development, we propose a model in which lineage commitment occurs independently of TCR gene rearrangement. PMID- 9394824 TI - Hypermutation, diversity and dissemination of human intestinal lamina propria plasma cells. AB - In this work we have microdissected lamina propria plasma cells and used polymerase chain reaction and sequencing to investigate immunoglobulin (Ig) gene rearrangements and mutations in human intestine. In addition, specific primers were designed for individual Ig gene rearrangements to analyze the distribution of related B cell and plasma cell clones at different sites along the bowel. Confirming our earlier work, intestinal IgVH genes were highly mutated in plasma cells from older individuals (> 30 years). IgVH genes were significantly less mutated in samples taken from patients aged 11-30 years, and there were fewer mutations again in samples from young children (< 11 years). In age-matched specimens the number of mutations was equivalent in the duodenum and colon. Using complementarity-determining region 3 primers to amplify specific Ig gene rearrangements, evidence was also found for the existence of related lamina propria plasma cells along the small bowel and colon, although these were quite scarce. In addition, analysis of the numbers of related clones in a random sampling from discrete areas of lamina propria indicates that the local population is diverse. These results suggest that the highly mutated IgVH genes in adult intestinal plasma cells are a consequence of chronic antigen exposure with age. Duodenal plasma cells are as highly mutated as colonic plasma cells, despite the fact that the upper bowel has no indigenous microbial flora (the stimulus for intestinal plasma cells). They also show that the plasma cell population is diverse and can be widely disseminated along the bowel. PMID- 9394825 TI - Expression and function of NKRP1A molecule on human monocytes and dendritic cells. AB - In this study, we analyzed the expression and function of the lymphocyte surface lectin NKRP1A on peripheral blood monocytes (Mo) or Mo and dendritic cells (DC) derived from thymic and bone marrow precursors. De novo expression of NKRP1A and CD14 molecules was detected upon culture of CD2- CD3- CD14- CD16- CD1a- NKRP1A- immature thymic precursors for 7 days in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF). Under these culture conditions, by day 21, a fraction of cells had lost CD14 and acquired both CD80 (B7.1) and CD86 (B7.2) molecules. These cells displayed a DC-like morphology and were surface NKRP1A positive. CD34+ NKRP1A- CD14- precursors, isolated from bone marrow and cultured in the presence of GM-CSF, also expressed both NKRP1A and CD14: these antigens were newly expressed on about one third of cells which had lost the CD34 precursor marker. In addition, NKRP1A was constitutively present on resting CD14+ peripheral blood Mo. When these cells were cultured in the presence of GM-CSF, the resulting DC population retained the expression of NKRP1A and acquired CD80, while they lost the CD14 antigen. Functional analysis revealed that the engagement of NKRP1A molecule leads to a strong intracellular calcium ([Ca2+]i) increase both in resting peripheral blood Mo and in vitro-derived DC. [Ca2+]i increase was mainly due to extracellular calcium influx, as it was completely abrogated by the addition of EGTA. More importantly, the engagement of the NKRP1A molecule induced interleukin (IL)-1 beta and IL-12 production by resting Mo and DC, respectively. Altogether these data indicate that NKRP1A lectin is present at the surface of Mo and DC and may play a relevant role in the activation and function of both cell types. PMID- 9394826 TI - Analysis of the tyrosine phosphorylation and calcium fluxing of human CD6 isoforms with different cytoplasmatic domains. AB - CD6 is a cell surface glycoprotein that functions both as a co-stimulatory and adhesion receptor on T cells. Recently we have described CD6 isoforms (CD6a, b, c, d, e) that arise via alternative splicing of exons encoding the cytoplasmic region of the molecule. CD6 becomes phosphorylated on tyrosine (Tyr) residues following stimulation through the T cell receptor (TCR) complex. Since the phosphorylation of Tyr residues renders some cell surface receptors competent for interactions with proteins of intracellular signaling pathways, we wanted to determine which region(s) and residues in the cytoplasmic domain of CD6 were important for phosphorylation on Tyr residues. We engineered and stably expressed chimeric receptors that consisted of the extracellular region of mouse CD6 and the cytoplasmic regions of either naturally occurring isoforms of human CD6, truncated proteins, or point mutants. We were able to demonstrate that of the nine Tyr residues in the cytoplasmic domain of the largest isoform CD6a, the two C-terminal Tyr residues (Tyr 629/662) are critical for the phosphorylation of CD6 following TCR cross-linking. Isoform CD6e, which is missing a region that contains two proline-rich motifs, is not phosphorylated. We further analyzed the ability of the different CD6 isoforms and truncated receptors to mobilize intracellular calcium after CD6/TCR co-ligation. All CD6 isoforms, including CD6e, as well as the truncation mutant delta 555, which is missing approximately the C-terminal half of the cytoplasmic domain, are able to increase Ca2+ influx. Taken together, these results suggest that the region of CD6 which is critical for Ca2+ mobilization is located N-terminal from amino acid 555 and is therefore different from the region located at the C terminus of CD6, which is necessary for tyrosine phosphorylation. PMID- 9394827 TI - Identification of a homolog of the C alpha 3'/hs3 enhancer and of an allelic variant of the 3'IgH/hs1,2 enhancer downstream of the human immunoglobulin alpha 1 gene. AB - Although four regulatory elements are known downstream of the mouse IgH alpha gene, a single enhancer homologous to hs1,2 has been thus far described downstream of each human alpha gene (Chen, C. and Birshtein, B. K., J. Immunol. 1997. 159: 1310). We characterized a 10-kb region downstream of the human alpha 1 gene. Two B cell-specific regulatory elements homologous to the murine C alpha 3'/hs3 and hs1,2,3' enhancers were found, which are duplicated downstream of alpha 2. The hs1,2 element is in inverted orientation by comparison with a recently reported alpha 1 hs1,2 element: it appears as a common allelic variant carrying an internal tandem repeat insertion and its prevalence in the human population is 60%. As in the mouse, the human hs1,2 enhancer is flanked with long inverted repeats which may have promoted inversion events through homologous recombination. Although the palindromic organization of the region is maintained in human, sequence identity with rodents focuses on core enhancer elements rather than on flanking repeats. Concerted divergence of both sides of the dyad symmetry suggests that inverted repeats are not just evolutionary remnants but rather play an architectural role in the LCR function. PMID- 9394828 TI - Thymic T cell export is not influenced by the peripheral T cell pool. AB - The peripheral T cell pool is maintained both by export of naive T cells from the thymus and by post-thymic expansion of activated/memory T cells. However, it is not known whether the thymus can alter its output following peripheral T cell depletion. Using intrathymic injection of fluorescein isothiocyanate to detect recent thymic emigrants (RTE), we directly tested whether the thymus is able to alter the number of RTE or the CD4:CD8 ratio of RTE emigrating to the periphery in response to in vivo depletion of total peripheral T cells or CD4 T cells, respectively. Depletion of peripheral T cells was achieved with anti-Thy-1 or anti-CD4, at doses that did not affect thymocyte numbers. Depletion of greater than 70% of peripheral T cells by treatment with anti-Thy-1 in vivo did not alter the number or cell cycle status of RTE trafficking to lymph nodes or spleen during the peripheral reconstitution phase (6, 9, 12 days). Similarly, depletion of the majority of CD4 T cells, which significantly reduced the peripheral CD4:CD8 T cell ratio, did not alter the total number or the proportion of CD4+ CD8- RTE in peripheral lymphoid organs. These data clearly indicate that thymic output is not influenced by downstream alterations in peripheral T cell pool size or CD4:CD8 ratio. Rather we contend that thymic T cell export is internally regulated by as yet undefined mechanisms. PMID- 9394829 TI - Immune complexes are potent inhibitors of interleukin-12 secretion by human monocytes. AB - We have studied the effect of immune complexes (IC) on interleukin (IL)-12 secretion by human monocytes in vitro. Two experimental models of IC were used. IC formed of tetanus toxoid and polyclonal anti-tetanus toxoid antiserum as well as heat-aggregated human serum IgG almost completely inhibited IL-12 (p70 and p40) secretion induced by interferon-gamma and lipopolysaccharide in human blood derived monocytes. Neutralizing anti-IL-10 antibodies plus indomethacin restored IL-12 secretion in the presence of IC to a high extent, indicating that IL-10 and prostaglandin (PG) partially mediate the IC-induced inhibition of IL-12 secretion. However, neutralization of tumor necrosis factor (TNF)-alpha by specific antibodies also incompletely restored IL-12 secretion. Indeed, monocytes secrete high levels of TNF-alpha upon stimulation by IC. We found that exogenously added TNF-alpha caused a profound inhibition of monocytic IL-12 secretion in the absence of IC, again mediated via the induction of IL-10 and PG. In summary, IC inhibit IL-12 secretion via TNF-alpha-induced IL-10 and PG synthesis. We conclude that IC, typically appearing in the course of chronic inflammatory processes, may influence the balance between Th1 and Th2 responses and may thus contribute to a deprivation of cell-mediated immune responses. PMID- 9394830 TI - Interactions between membrane IgM and the cytoskeleton involve the cytoplasmic domain of the immunoglobulin receptor. AB - Cross-linking induced interactions between the membrane form of immunoglobulin (mIg) and the cytoskeletal matrix have been described by several groups. To date, the function of mIgM association with the cytoskeleton is not yet understood. Delineation of the molecular basis of these interactions will be instrumental in elucidating their function. We have previously shown that the Ig alpha/beta heterodimer is not required for ligand-induced mIgM binding to the cytoskeleton. In this study, we have investigated the role of other B cell-specific proteins in mediating these interactions. For this, we expressed mIgM in the non hematopoietic human cervical carcinoma cell line HeLa S3 and verified the capacity of the surface-expressed IgM to interact with the cytoskeletal matrix upon cross-linking with anti-mu chain antibodies. We show here that only the mIgM molecule itself and no other B cell-specific protein(s) is required in mediating mIgM interactions with actin filaments. In an attempt to determine the cytoskeleton-binding site of mIgM we investigated the role of the cytoplasmic tail of mIgM (KVK) in binding the receptor to actin-based microfilaments. Using mutated forms of mIgM expressed in J558L cells, we show here that KVK plays a role in mediating these interactions. The absence of KVK did not, however, completely abrogate mIgM-cytoskeletal interactions, suggesting that there are additional molecular requirements for the ligand-induced mIgM binding to the cytoskeletal matrix. PMID- 9394831 TI - Analysis of immunoreceptor tyrosine-based activation motif (ITAM) binding to ZAP 70 by surface plasmon resonance. AB - The signaling function of the T cell antigen receptor (TCR) is mediated via CD3 polypeptides, the cytoplasmic sequences of which bear conserved immunoreceptor tyrosine-based activation motifs (ITAM). ITAM are defined by two YxxL/I sequences separated by a six-eight amino acid long spacer. Upon antigen recognition, ITAM become phosphorylated on both tyrosine residues, creating a high affinity binding site for the tandem SH2 domains found in the protein tyrosine kinase ZAP-70. Using surface plasmon resonance, we further dissected the sequences required for the binding of ZAP-70 to each TCR-associated ITAM. First, we generated protein tyrosine phosphatase-resistant ITAM peptide analogs, in which difluorophosphonomethyl phenylalanyl (F2p) replaced both phosphotyrosines, and showed that those protein tyrosine phosphatase-resistant analogs bind ZAP-70 with high affinity, establishing a rational strategy for the design of novel pharmacological tools capable of interfering with TCR signaling function. Second, we substituted the five amino acids separating the two YxxL/I sequences of the CD3 zeta 1 ITAM with a non-peptidic linker made up of gamma-amino butyric acid units and demonstrated that the length of this intervening sequence rather than its chemical composition is essential for high affinity binding of phosphorylated ITAM to the ZAP-70 SH2 domains. PMID- 9394832 TI - Thiol modulation inhibits the interleukin (IL)-1-mediated activation of an IL-1 receptor-associated protein kinase and NF-kappa B. AB - The interleukin-1 receptor type I (IL-1RI) is associated with other proteins thus forming a complex system by which IL-1 exerts its various signals. The initiating event is still uncertain, but activation of a recently described receptor associated protein kinase is one of the earliest events detectable (Martin et al., Eur. J. Immunol. 1994. 24: 1566). IL-1 signaling is commonly accompanied by oxidative processes and is thought to be subject to redox regulation. We therefore investigated whether the activation of the IL-1RI-associated protein kinase could be a target for redox regulation and whether an altered activity of the kinase could influence IL-1-mediated NF-kappa B activation. A murine T cell line, EL4, was stimulated with IL-1 with and without pretreatment with different compounds known to influence the cellular redox status. Thiol modifying agents like diamide, menadione, pyrrolidine dithiocarbamate (PDTC), diethyl dithiocarbamate or phenylarsine oxide inhibited the IL-1-induced activation of the IL-1RI-associated protein kinase. N-Acetylcysteine, alpha,alpha'-dipyridyl, aminotriazole or nitrofurantoin did not show any effect. The inhibition by PDTC was reversible unless glutathione synthesis was blocked by buthionine sulfoximine. The described conditions which inhibited or prevented the activation of the IL-1RI-associated kinase similarly impaired the activation of NF-kappa B in EL4 cells. From these observations we conclude that free thiols in the IL-1RI complex are essential for the activation of the IL-1RI-associated protein kinase and that this process is mandatory for IL-1 signaling leading to NF-kappa B activation. PMID- 9394833 TI - Molecular mechanisms regulating induction of interleukin-6 gene transcription by interferon-gamma. AB - The multifunctional cytokine interleukin-6 (IL-6) plays a central role in host defence mechanisms and hematopoiesis. Furthermore, dysregulation of IL-6 gene expression is associated with the pathogenesis of various immunologically related diseases such as myeloma, systemic lupus erythematosus, rheumatoid arthritis, psoriasis and Kaposi's sarcoma. The regulation of IL-6 gene expression occurs mainly at transcriptional level, although mechanisms of post-transcriptional regulation have also been described. In the present study we demonstrate that in HeLa cells, induction of IL-6 by interferon-gamma (IFN-gamma) is transcriptionally controlled, as shown by run on assays and analysis of the IL-6 mRNA stability. Gel-retardation experiments using antibodies specific for factors of the IRF family identified four protein-DNA complexes, which bind to the interferon regulatory factor (IRF) binding site at position -267 to -254, in nuclear extracts from IFN-gamma treated cells. Furthermore, transient transfection analyses of the 5'-flanking region of IL-6 gene linked to the chloramphenicol acetyltransferase (CAT) reporter gene demonstrated that the -267 to -254 IRF site is necessary for IL-6 induction by IFN-gamma. However, transfection experiments in which IRF-1 and I kappa B alpha were overexpressed show that full-scale transcriptional activation of the IL-6 promoter directing CAT expression requires the co-operation between IRF-1 and NF-kappa B at a low constitutive level. PMID- 9394834 TI - Dendritic cells differently respond to haptens and irritants by their production of cytokines and expression of co-stimulatory molecules. AB - After application of haptens to the skin, Langerhans cells (LC), i.e. immature dendritic cells (DC) in the skin, move to secondary lymphoid organs to sensitize naive T cells. During this process, LC become mature DC with augmented expression of various co-stimulatory molecules and MHC class II antigens. In this scenario, however, critical questions remain as to what kind of chemicals can induce this maturation process through what kind of mechanisms. To clarify these questions, we used monocyte-derived CD1a+ DC instead of LC since LC maturated spontaneously in vitro culture. After we confirmed that monocyte-derived DC showed at least phenotypic characteristics and a response to TNF-alpha similar to LC, we added various chemicals, i.e., dinitrochlorobenzene (DNCB), trinitrochlorobenzene (TNCB), NiCl2, ZnCl2, sodium dodecyl sulfate (SDS), or benzalkonium chloride (BC), to a culture of purified monocyte-derived CD1a+ DC. Of these chemicals, only NiCl2 and DNCB significantly increased the surface expression of CD54, CD86, HLA-DR antigen, and interleukin (IL)-1 beta production, while SDS, BC, or ZnCl2 could not augment them, except for weak augmentation of CD86 expression by SDS. The increase in the expression of CD86 induced by NiCl2 or DNCB was most remarkable, being observed in DC from almost all the subjects we examined. TNCB could also induce responses similar to those induced with DNCB, but the number of subjects whose DC responded to it was far less than that of subjects whose DC responded to NiCl2 or DNCB. In spite of the augmented CD86 expression on DC treated with DNCB or NiCl2, these chemicals induced different responses of DC in their expression of CD54 and HLA-DR and the production of IL-6 and tumor necrosis factor (TNF)-alpha. In addition, the up-regulation of CD86 expression on DC treated with DNCB was significantly suppressed by either anti-IL-1 beta or anti TNF-alpha antibody, while that by NiCl2 was relatively insensitive to these antibody treatments. Finally, the protein kinase C inhibitor, H7, but not staurosporine, could suppress the augmentation of CD86 expression on DC induced either by NiCl2 or by DNCB. These data suggest that DC respond to some haptens by changing their expression of several co-stimulatory molecules and their production of cytokines with a resultant change in antigen-presenting function. They also suggest that these chemicals stimulate DC by different mechanisms. By these responses, DC may modulate the final immune response to chemicals. PMID- 9394835 TI - Separately expressed T cell receptor alpha and beta chain transgenes exert opposite effects on T cell differentiation and neoplastic transformation. AB - Two aspects of T cell differentiation in T cell receptor (TCR)-transgenic mice, the generation of an unusual population of CD4-CD8-TCR+ thymocytes and the absence of gamma delta cells, have been the focus of extensive investigation. To examine the basis for these phenomena, we investigated the effects of separate expression of a transgenic TCR alpha chain and a transgenic TCR beta chain on thymocyte differentiation. Our data indicate that expression of a transgenic TCR alpha chain causes thymocytes to differentiate into a CD4-CD8-TCR+ lineage at an early developmental stage, depleting the number of thymocytes that differentiate into the alpha beta lineage. Surprisingly, expression of the TCR alpha chain transgene is also associated with the development of T cell lymphosarcoma. In contrast, expression of the transgenic TCR beta chain causes immature T cells to accelerate differentiation into the alpha beta lineage and thus inhibits the generation of gamma delta cells. Our observations provide a model for understanding T cell differentiation in TCR-transgenic mice. PMID- 9394836 TI - Conservation of a master hematopoietic switch gene during vertebrate evolution: isolation and characterization of Ikaros from teleost and amphibian species. AB - The generation of T, B and NK lymphocyte lineages from pluripotent hematopoietic stem cells is dependent upon the early expression of the Ikaros locus which by means of alternative splicing produces a variety of zinc finger DNA binding transcription factors. We assessed the general biological importance of Ikaros by studying its conservation and expression in teleost fish and amphibians. Portions of Ikaros cDNA from rainbow trout and Xenopus were amplified by reverse transcription-polymerase chain reaction (RT-PCR). They show roughly 75% conservation of the amino acid sequence with mammalian Ikaros. The trout fragment was then used to isolate full-length Ikaros clones from a trout thymocyte cDNA library. In mice and humans, Ikaros produces six alternatively spliced isoforms, but in trout two additional novel splice variants designated Ik-7 and Ik-8 were also found. Ik-7 is expressed in a similar fashion to Ik-1 and Ik-2, the predominant isoforms expressed in mammalian lymphocytes. In trout and zebrafish, as in mammals, Ikaros is a single-copy gene, but in Xenopus segregation analysis demonstrates that Ikaros has been duplicated, most likely a result of polyploidization. We then examined the expression of Ikaros in trout and Xenopus tumor T cell lines via Northern blot, RT-PCR, immunofluorescence and Western blot analysis. Overall, Ikaros is expressed in a lymphoid-specific fashion similar to that found in mice and humans. In addition Ikaros is expressed early in trout ontogeny, beginning roughly at days 3-4 in the yolk sac and at day 5-6 in the embryo proper. The conservation of Ikaros structure and expression confirms it as a master switch of hematopoiesis. PMID- 9394837 TI - Predominance of the autoimmune response to myelin oligodendrocyte glycoprotein (MOG) in multiple sclerosis: reactivity to the extracellular domain of MOG is directed against three main regions. AB - Our previous analysis of the T cell reactivity to myelin antigens in a group of 24 patients with multiple sclerosis (MS) and 16 control individuals revealed that the autoimmune response to myelin oligodendrocyte glycoprotein (MOG) predominates in MS over that to myelin basic protein (MBP), proteolipid protein or myelin associated glycoprotein, suggesting a prevalent role for the autoimmune response to MOG in the pathogenesis of MS. Using a recombinant human MOG (rhMOG) preparation corresponding to the extracellular immunoglobulin-like domain of the MOG molecule, we have now analyzed another group of 52 MS patients and 49 control individuals for reactivity of their peripheral blood lymphocytes (PBL) to rhMOG and to MBP concomitantly. Of the 52 MS patients tested 24 responded to MOG and 10 out of 49 responded to MBP, whereas only 5 MOG-reactive and 4 MBP-reactive control individuals were detected out of the 49 tested. These results are therefore highly confirmatory of the predominant reactivity to MOG in MS. The analysis of the primary proliferative response to 11 synthetic overlapping peptides (phMOG) spanning the extracellular domain of human MOG by PBL from 9 MS patients and 15 control individuals (9 healthy controls and 6 patients with neurological diseases other than MS) further supports a prevalent role for the autoimmune response to MOG in MS, as only 1 of the 15 controls tested showed reactivity to any of the phMOG, whilst 5 out of the 9 patients studied reacted to at least 1 of the phMOG. PBL from 10 MS patients, and from 4 controls, were selected in vitro with each of the phMOG. Of the 10 patients studied 7 reacted to at least 1 phMOG upon secondary stimulation and the reactivity was mostly directed to epitopes localized within three main regions (amino acids 1-22, 34-56 and 64-96), as was observed for the primary response of PBL. The predominant response to MOG of PBL from MS patients as demonstrated in two separate studies using native MOG and rhMOG as antigens, and the high incidence of reactivity of these PBL compared to the lack of response to phMOG by control PBL, emphasize the relevance of MOG in MS pathogenesis and support a primary role for the autoimmune T cell response to MOG in disease development. PMID- 9394839 TI - A T cell receptor (TCR) antagonist competitively inhibits serial TCR triggering by low-affinity ligands, but does not affect triggering by high-affinity anti-CD3 antibodies. AB - It has been demonstrated that modified peptides which fail to induce detectable T cell responses can act as T cell receptor (TCR) antagonists when presented together with agonist by the same antigen-presenting cell (APC). We report that a TCR antagonist competitively inhibits TCR triggering induced by low-affinity ligands such as agonistic peptides or bacterial superantigens. However, the same antagonist cannot inhibit TCR triggering and T cell activation induced by high affinity anti-CD3 antibodies that engage most TCR at once. These results indicate that TCR antagonists inhibit T cell responses by interfering with the ongoing process of serial triggering, rather than by delivering an inhibitory signal to T cells. PMID- 9394838 TI - Sequence analysis of rat integrin alpha E1 and alpha E2 subunits: tissue expression reveals phenotypic similarities between intraepithelial lymphocytes and dendritic cells in lymph. AB - The integrin alpha OX-62 subunit is defined by the OX-62 monoclonal antibody that was raised against rat dendritic cells in lymph (veiled cells) and shows properties similar to those of human alpha E2 that is predominantly expressed on intraepithelial lymphocytes. To clone alpha OX-62, rat probes generated using primers specific for the human alpha E sequence were used to screen rat T cell cDNA libraries. cDNA clones encoding two similar but not identical alpha subunits that are closely related to but distinct from human alpha E were isolated. alpha E1 is predicted to be the rat homolog of mouse alpha M290 and alpha E2 corresponds to rat alpha OX-62. Immunofluorescence analysis revealed that mouse alpha E1 and rat alpha E2 are expressed in dendritic epidermal T cells in the skin, intraepithelial lymphocytes in the small intestine and in cells with a dendritic morphology present at sites where gamma delta T cells occur in lymphoid organs. Unexpectedly, alpha E2 is co-expressed with intracellular CD3-delta and a 33-kDa CD3 chain but not the T cell receptor in veiled cells. These findings suggest that veiled cells may be derived from a lymphoid precursor. Furthermore, veiled cells show phenotypic similarities to intraepithelial lymphocytes. PMID- 9394841 TI - Medical oncology as a discipline. AB - Medical oncology has become a major subspecialty discipline of internal medicine within only 25 years. The special skill of oncologists is judgement in matters relating to cancer. This specialty brought the necessary expertise for cancer management into the community, improved care of patients with cancer worldwide, and provided a significant impact on cancer education and cancer research. Current manpower needs have been met but the ultimate in desired services are not apparent. Attempts to merge hematology and oncology are unwarranted. The future for medical oncology will be challenging, especially to meet the expanding needs of geriatric oncology. PMID- 9394840 TI - The low-grade lymphoproliferative disorders. AB - Rapid advances in our understanding of the biology and pathology of lymphoproliferative disorders, permitted mainly by new diagnostic tools, constantly change our approach to this heterogenous group of disorders. In this review of the more indolent subgroup of lymphoproliferative disorders, some of the recent advances are highlighted, and treatment options discussed. PMID- 9394842 TI - The relationship of shape of tumor invasion to depth of invasion and cervical lymph node metastasis in squamous cell carcinoma of the tongue. AB - Several investigators have suggested that there is a strong correlation between tumor depth and lymph node involvement in tongue carcinoma. The purpose of this study was to investigate the relationship between the shape of tumor, tumor depth, and lymph node metastasis in tongue carcinoma. Fifty-four patients with T1 abd T2 tongue carcinomas who underwent surgical treatment were included in this study. Tumors were divided into four categories according to their shape of invasion: superficial, exophytic, endophytic, and a combination of endophytic and exophytic. Forty cases of endophytic and combination types were divided into two groups according to the shape of invasion: (1) reductive bottom of invasion (n = 17) and (2) expansive bottom of invasion (n = 23). Tumors with a reductive bottom of invasion showed a variety of tumor depths and had low lymph node involvement (4/17, 23.5%). However, tumors with an expansive bottom of invasion showed deeper invasion and a high incidence of lymph node metastasis (16/23, 69.6%). These results suggested that the macroscopic shape of invasion is a feature that may provide important information about the prognosis of the primary tumor especially in relation to cervical lymph node involvement. PMID- 9394843 TI - A case-control study on risk factors for local recurrences or distant metastases in breast cancer patients treated with breast-conserving surgery. AB - A case-control study was conducted on 143 case-control sets recruited from the 18 key hospitals/ institutes in Japan to identify risk factors for local recurrences and distant metastases after breast-conserving surgery in breast cancer patients: (1) positive surgical margin was a risk factor for local recurrences (relative risk (RR) = 16.70, p < 0.001) but not for distant metastases, (2) positive p53 immunostaining was a risk factor for both local recurrences (RR = 5.62, p = 0.003) and distant metastases (RR = 3.11, p = 0.034), (3) lymph node metastasis was a risk factor for distant metastases (RR = 9.28, p = 0.001) but not for local recurrences, (4) radiation therapy reduced local recurrences (RR = 0.13, p = 0.004) and (5) adjuvant chemotherapy (RR = 0.27, p = 0.120), endocrine therapy (RR = 0.21, p = 0.037), and chemoendocrine therapy (RR = 0.05, p = 0.013) reduced local recurrences. PMID- 9394844 TI - Tissue expression and serum levels of HER-2/neu in patients with breast cancer. AB - We have analyzed serum levels of soluble HER-2/neu in 42 primary breast cancer patients prior to any therapy and studied the relationship between the overexpression and amplification of HER-2/neu in the primary tumor after surgical excision and data obtained by pathohistological staging. In addition, we have investigated the sera of 62 patients with stage IV breast cancer. Using an enzyme linked immunosorbent assay, we observed elevated serum HER-2/neu levels in 6/42 (14.2%) preoperative patients. In 42.8% of the patients with HER-2/neu tumor expression/amplification serum levels were increased. In contrast, only 8.5% of the patients without HER-2/neu expression/amplification in the primary tumor presented with elevated serum levels. There was a significant correlation between serum concentrations of soluble HER-2/neu and tumor size (p < 0.0001) or axillary lymph node involvement (p < 0.0001). In patients with stage IV disease, 27 of 62 (43.5%) had elevated soluble HER-2/neu serum levels. A highly significant correlation between soluble HER-2/ neu and CA 15-3 (p < 0.002) was observed. The correlation of serum concentrations of HER-2/neu with estrogen and progesterone receptor status of the primary tumor was not significant in both groups. In conclusion, the measurement of serum HER-2/neu levels at diagnosis defines a small subgroup of breast cancer patients with a relatively advanced stage of disease. Its strong correlation with tumor load in patients with stage II disease and the high prevalence in patients with stage IV disease could make it a promising tool for the assessment of disease activity and biologic behavior in breast cancer. PMID- 9394845 TI - Relationship of serum testosterone level with proliferating cell nuclear antigen and nm23 protein in human prostatic carcinoma tissue. AB - OBJECTIVE: To elucidate the biological significance of proliferating cell nuclear antigen (PCNA) and nm23 immunoreactivity in prostatic carcinoma (PC) tissue, both expressions were immunohistochemically analyzed, and the results were compared with the change of the serum testosterone (T) level. METHODS: The paraffin embedded materials obtained from 49 untreated PC and 16 hormonally refractory PC (hr-PC) were used. Of the 49 untreated PC, 35 received luteinizing hormone releasing hormone (LH-RH) analogue treatment, while 14 received a cisplatin-based chemotherapy. The immunohistochemistry of PCNA and nm23 protein was performed using an anti-PCNA monoclonal antibody (PC-10) and an antihuman nm23 polyclonal antibody (OA-11-890), respectively. The serum T level was measured by means of radioimmunoassay. RESULTS: In both untreated PC and hr-PC, the immunoreactivity of nm23 protein significantly correlated with the PCNA expression. Both PCNA expression and nm23 protein immunoreactivity were higher in poorly differentiated PC than those observed in well-differentiated PC, while no significant difference in the serum T level was observed between poorly and well-differentiated PCs. On the other hand, both PCNA expression and nm23 protein immunoreactivity were significantly higher in hr-PC than those observed in untreated PC, whereas the serum T level was significantly lower in hr-PC. In 35 PCs treated with LH-RH analogue, no significant difference in both PCNA expression and nm23 protein immunoreactivity was found between those specimens obtained before and at 3 months after the treatment, while a significant reduction of the serum T level was noted at 3 months after the treatment. Similarly, in 14 PCs treated with a cisplatin-based chemotherapy, the same change of PCNA expression and nm23 protein immunoreactivity as observed in LH-RH analogue treatment was found, while no significant difference of the serum T level was found. CONCLUSIONS: These findings appear to indicate that (1) nm23 protein immunoreactivity is interrelated with cellular proliferation in PC tissue and (2) alteration of the serum T level during a short period was not enough to explain the essential change of cellular proliferation of PC tissue, but might reflect other aspects of tumor growth such as apoptosis. PMID- 9394846 TI - Adrenocortical carcinoma: experience in 45 patients. AB - Forty-five patients with adrenocortical carcinoma (13 nonfunctioning and 32 functioning carcinomas) were retrospectively studied. Five-year survival rate was 29% overall; for patients at stage I-II (n = 15) it was 70%, and for patients at stage III-IV (n = 30) it was 12%. In patients given mitotane + chemotherapy survival rate was similar to that observed in patients given chemotherapy alone, and significantly longer than in patients given mitotane alone (p < 0.05). There were no differences in disease-free interval and survival between adjuvant mitotane and no treatment. Optimization of therapeutic protocols in addition to early recognition may improve prognostic aspects of this type of malignancy for which treatment outcome is still unsatisfactory. PMID- 9394847 TI - Serum steroids in relation to benign prostatic hyperplasia. AB - In order to investigate the endocrine etiology of benign prostatic hyperplasia (BPH), we conducted a case-control study in Athens, Greece using 52 patients with histologically confirmed BPH and 52 healthy controls matched according to age and town of residence. Blood samples were collected from all participants and analyzed blindly in Boston, Mass. regarding testosterone (T), dihydrotestosterone, estradiol (E2), sex hormone-binding globulin, and dehydroepiandrosterone sulfate (DHEAS). Results from logistic regression models, adjusting for age, height, body mass index, years of schooling, and mutually among the measured hormones, indicate that DHEAS is significantly positively associated with the risk of BPH [odds ratio = 3.10 per standard deviation (60 micrograms/dl), 95% confidence interval (1.28,7.50)]. T and E2 were not significantly related to the risk of BPH. PMID- 9394848 TI - Relaxation of insulin-like growth factor-II gene imprinting in human gynecologic tumors. AB - To test for the existence of genomic imprinting in human gynecologic tumors, we analyzed the allelic expression of human insulin-growth factor-II (IGF-II) genes. Genomic imprinting is the parental allele-specific expressions of genes, and recently imprinting of IGF-II gene has demonstrated that parental IGF-II was monoallelically expressed. To study whether IGF-II gene imprinting occurs in human gynecologic tumors, we examined allele-specific expression using an ApaI polymorphism in the 3' untranslated region of IGF-II gene exon 9. We used 19 gynecologic tumor cell lines, and 66 human gynecologic tumors. Four of 19 cell lines (21%) were informative, and three of these four cell lines (75%) revealed loss of imprinting (LOI). For gynecologic tumors, 24 of 66 were informative (36%), and 5 of the 24 (21%) had LOI. We have reported here that the IGF-II gene is expressed biallelically in some gynecologic tumors. We suggest that LOI of the IGF-II gene is involved in the development of some gynecologic tumors. PMID- 9394849 TI - Expression of p16 and cyclin-dependent kinase 4 proteins in primary breast carcinomas. AB - The immunolocalization of the p16 and cdk4 proteins was investigated in 65 retinoblastoma gene product (pRB)-positive and 20 pRB-negative breast carcinomas. These proteins were expressed in similar lesions in 84.6% of the pRB-positive and 100% of the pRB-negative carcinomas. Diffuse expression of p16 was observed in 73.8 and 70.0% of the pRB-positive and -negative cases, respectively. cdk4 and p16 expression was significantly more heterogeneous in tumors of larger sizes and/or at higher stages. These findings suggest that p16 can be induced regardless of pRB status and Rb gene function in primary breast carcinoma and that it modulates the cell cycle progression in association with cdk4. PMID- 9394850 TI - Survival with chronic myelogenous leukemia. PMID- 9394851 TI - Octreotide for the treatment of hypercalcemia related to B cell lymphoma. PMID- 9394852 TI - A review of endocrine options for the treatment of advanced breast cancer. AB - It is now 100 years since it was first recognised that altering the endocrine environment can be valuable in patients with inoperable breast cancer. Various ablative endocrine and other treatments have since been developed, but few have resulted in a substantial improvement in clinical efficacy, and the response rates to endocrine manoeuvres remain at about 30%. The benefits of more recent therapies, such as the new generation of aromatase inhibitors, appear to relate to better tolerability and more convenient administration. It is now recommended that the majority of patients with advanced breast cancer should receive endocrine therapy as their first mode of treatment, although it is postmenopausal patients who are likely to gain most benefit from this approach. Tamoxifen is currently the first choice of treatment in the majority of postmenopausal women, although most eventually experience progression of disease, possibly because of the partial oestrogen agonist activity of tamoxifen. Second-line therapy is usually then either an aromatase inhibitor or a progestin. The new-generation selective, non-steroidal aromatase inhibitors are effective and can offer major clinical benefits in terms of fewer side-effects. PMID- 9394853 TI - The relevance of preclinical models to the treatment of postmenopausal breast cancer. AB - The predictive value of test results in animals when selecting a compound for potential therapeutic human use depends upon the relevance of the animal model to the human disease and the comparative pharmacokinetics of the compound in animals and man. The development of the aromatase inhibitor, anastrozole (Arimidex), illustrates the importance of these factors. In postmenopausal women with breast cancer, aromatase activity in the peripheral tissues is the main source of oestrogen for tumour growth. Only one form of the human enzyme is known, which is not subject to strong feedback control. Inhibition of aromatase therefore simply reduces oestrogen production. This situation is mimicked by assays of acute inhibition of ovulation in rats, chronic inhibition of androstenedione-induced uterine hypertrophy in sexually immature rats, and chronic inhibition of peripheral aromatase in monkeys. In all these assays, maximum anastrozole activity was consistently achieved at an oral dose of about 0.1 mg/kg, and the clearance half-life of 7-16 h indicated that once-daily dosing would be possible in humans. The clearance half-life in postmenopausal women is about 50 h, and with once-daily dosing the dose of anastrozole required for maximal inhibition is 1 mg/day. The rat 7,12-dimethylbenzanthracene tumour model, in contrast, is supported by ovarian oestrogen and chronic inhibition provokes positive feedback loops that try to restore oestrogen production, masculinise the animals and decrease the clearance half-life of anastrozole. Higher doses (10 mg/kg) of anastrozole are therefore needed. Variations in the dose of aromatase inhibitor required in different models, therefore, can be explained in terms of pharmacokinetics and do not reflect the effectiveness of anastrozole as an aromatase inhibitor. PMID- 9394854 TI - Anastrozole--a new generation in aromatase inhibition: clinical pharmacology. AB - Use of the aromatase inhibitor aminoglutethimide is limited by its lack of selectivity for aromatase and its toxicity. Newer agents are more selective, but do not always offer improved inhibition of aromatase. Indirect comparison of their activity in inhibiting aromatase and suppressing plasma oestrogens indicates that aminoglutethimide, rogletimide, formestane, and fadrozole inhibited aromatase activity by 74-91%, with reported falls in oestradiol level of 58-76%. In contrast, the new-generation oral once-daily aromatase inhibitors anastrozole (Arimidex) and letrozole were of a similar activity, inhibiting aromatase activity by over 96%, with a concomitant fall in oestradiol and oestrone levels of at least 80%. Anastrozole at the recommended clinical dose of 1 mg daily also suppressed oestrone sulphate levels by 93.5%; activity with anastrozole 10 mg daily was not statistically significantly different. The new generation of aromatase inhibitors, as typified by anastrozole, thus offers effective and convenient aromatase inhibition which correlates well with decreases in the levels of plasma oestrogens. PMID- 9394855 TI - Clinical overview of anastrozole--a new selective oral aromatase inhibitor. AB - The efficacy and tolerability of the new selective aromatase inhibitor, anastrozole (Arimidex), was compared with megestrol acetate in the treatment of advanced breast cancer in postmenopausal women. In two independent prospective randomised trials, patients who progressed after prior tamoxifen therapy received anastrozole 1 or 10 mg once daily, or megestrol acetate, 40 mg q.i.d. The two studies were designed to allow the data to be combined to increase the statistical power of the analyses. It is the data from these combined analyses that are considered in further detail. After 6 months of follow-up, the proportion of patients gaining clinical benefit (complete response + partial response + stable disease > or = 24 weeks) was approximately one third of patients in all three groups. No significant difference was observed between either dose of anastrozole and megestrol acetate in the time to disease progression (130-153 days). All three treatments were generally well tolerated, but significantly more patients on megestrol acetate gained weight, and the weight gain in this group continued up to at least 9 months of follow-up. At a 12 month update of tolerability, of the commonly observed adverse events, a greater than 2-fold difference between treatment arms was observed for hypertension, weight gain, dyspnoea, vaginal haemorrhage, sweating and diarrhoea (all higher on megestrol acetate except for diarrhoea). Anastrozole is effective and well tolerated and on the basis of these data, 1 mg once daily is the recommended clinical dose in postmenopausal women with advanced breast cancer. PMID- 9394856 TI - A randomised comparison of oestrogen suppression with anastrozole and formestane in postmenopausal patients with advanced breast cancer. AB - The relative efficacy and tolerability of the aromatase inhibitors anastrozole (Arimidex) and formestane are assessed in a direct comparative trial in postmenopausal women with advanced breast cancer. Final results are available and reported here only for oestradiol suppression. Patients were randomised to receive either oral anastrozole, 1 mg once daily, or formestane, 250 mg every 2 weeks intramuscularly. In the anastrozole group, mean serum oestradiol levels fell from 32.1 pmol/l at baseline to 6.5 pmol/l at week 1, and similar levels of suppression were maintained over the next 3 weeks. In the formestane group, mean serum oestradiol levels fell from 31.0 pmol/l at baseline to 9.5 pmol/l at the week 1 assessment. In this group, serum oestradiol levels tended to rise by the 2 and 4-week measurements, i.e. immediately before the next injection was due. Based on the 2- and 4-week measurements, the mean falls in oestradiol levels were 79 and 58% in the anastrozole and formestane groups, respectively (p = 0.0001). More effective and consistent suppression of oestradiol was achieved with anastrozole at the therapeutic dose of 1 mg once daily, orally, than with formestane at the standard dose of 250 mg every 2 weeks, intramuscularly. PMID- 9394857 TI - Tolerability of endocrine treatment for advanced breast cancer: results of an international survey. AB - In contrast with cytotoxic therapy, the impact of hormonal therapy for advanced breast cancer in postmenopausal women is poorly documented. Two recent surveys of the impact of hormonal therapy in this patient group found that side-effects had an adverse impact on patients' quality of life which needed to be taken into account when discussing treatment options. Patients were perceived to be more comfortable discussing treatment-related problems with nurses rather than doctors, who tended to underestimate the distress caused to patients by the side effects of treatment. A need was identified for better lines of communication between patients and medical professionals, and for a simple, more widely applicable tool for assessment of side-effects. As a result of these findings, a simple checklist, Checklist for Patients on Endocrine Therapy (C-PET), has been developed to be completed by patients before a physician appointment, to form a basis for discussion. A pilot study has indicated that C-PET is easy and quick for patients to complete and aids communication between patients and healthcare professionals. C-PET has now been formally launched to nurses, oncologists, and other cancer physicians. PMID- 9394858 TI - The role of selective non-steroidal aromatase inhibitors in future treatment strategies. AB - The major role of endocrine therapy in the treatment of early and advanced breast cancer should not be forgotten as an increasing number of new and potentially more exciting agents are introduced. Endocrine therapy generally has a more powerful effect than cytotoxic therapy in the treatment of breast cancer. Alternatives to tamoxifen, such as anastrozole (Arimidex), are now being considered, both in advanced disease and as adjuvant therapy, whether as single agents or in combination. Anastrozole has a low side-effect profile and is becoming a drug of choice for second-line therapy in postmenopausal women with advanced breast cancer. The present use of anastrozole in postmenopausal women with advanced breast cancer includes second-line treatment after tamoxifen and first-line treatment after tamoxifen has been used as an adjuvant treatment. It may also be used when tamoxifen is not tolerated. The future potential of anastrozole is currently being investigated in a programme of clinical trials in postmenopausal women, including first-line treatment of advanced disease and as an adjuvant treatment for early breast cancer. PMID- 9394859 TI - Cerebral blood flow and energy metabolism in the newborn. AB - In normal newborn term and preterm infants CBF is relatively low corresponding to a low metabolic rate for oxygen, whereas cross-brain oxygen extraction is similar to that in adults. This provides for a considerable reserve capacity to deal with decreased CBF or decreased oxygen content in arterial blood. CBF reactivity to CO2 is normal, and the evidence is that pressure-flow autoregulation is present, even in very preterm infants. Absence of autoregulation and CBF-CO2 reactivity has been documented in severely asphyxiated infants, and in preterm infants who went on the develop severe intracranial hemorrhage. A number of methods are available to study CBF and brain metabolism in newborn infants. Several of them involve ionizing radiation, which has limited their use, even though it is unlikely that the associated risks are particularly high. Magnetic resonance spectroscopy has demonstrated a delayed disturbance of energy metabolism following severe asphyxia. Doppler ultrasound has rarely been helpful to obtain quantitative data. Near infrared spectrocopy has now been in use for more than 10 years. It has been slow to fulfill its promise as a continuous monitor of cerebral circulation and of oxygen sufficiency of neurons. PMID- 9394860 TI - Cerebral hemodynamics and oxygenation in the fetus. The role of intrapartum near infrared spectroscopy. AB - Current methods of intrapartum surveillance have made little impact on fetal mortality and morbidity while leading to increased caesarean section rates. Near infrared spectroscopy is a powerful new technique that can continuously measure changes in fetal cerebral oxygenation and hemodynamics during labor. Data are presented suggest that near-infrared spectroscopy has potential as a new form of fetal monitoring. However, further technical developments and testing in clinical trials are necessary before its introduction into clinical practice. PMID- 9394861 TI - Brain injury in the premature infant. Neuropathology, clinical aspects, pathogenesis, and prevention. AB - There are two principal lesions that underlie brain injury and the neurologic manifestations in the premature infant: periventricular hemorrhagic infarction and periventricular leukomalacia. Both of these lesions may be potentially preventable: periventricular hemorrhagic infarction by preventing germinal matrix IVH, and periventricular leukomalacia by detecting impaired cerebrovascular regulation with near-infrared spectroscopy, preventing the impaired cerebral blood flow and interrupting the cascade to oligodendroglial cell death, perhaps by such agents as free-radical scavengers. Prenatal magnesium sulfate also may be valuable. The greatest progress toward prevention has been made regarding periventricular hemorrhagic infarction, but the advent of new technologies, especially near-infrared spectroscopy, and of new insights into the cellular basis for oligodendroglial vulnerability provide hope for prevention of periventricular leukomalacia. PMID- 9394862 TI - Intraventricular hemorrhage and posthemorrhagic hydrocephalus. Current and potential future interventions. AB - Enhanced survival of very premature infants may be regarded as the most striking demonstration of the major improvements in perinatal medicine during the last two decades. This article discusses recent perinatal interventions in the context of the pathogenesis and know risk factors for intraventricular hemorrhage (IVH). In addition, controversies related to the evolution and management of posthemorrhagic hyrdocephalus (PHH) are examined, and current concepts concerning potential brain injury to PHH are reviewed. PMID- 9394863 TI - Hypoxic-ischemic brain injury in the term newborn. Neuropathology, clinical aspects, and neuroimaging. AB - Hypoxic-ischemic cerebral injury in the full-term infant results in a variety of neurologic manifestations. The pathogenetic events resulting in this central nervous system injury may occur throughout the prenatal period. Several clinical patterns of signs and symptoms of hypoxic-ischemic cerebral injury have been identified in the term infant. Further, characteristic neuroradiologic patterns of this injury can be discerned. Information derived from the term infant's clinical course and neuroimaging data convey useful neurodevelopmental prognostic information. Several potential and promising therapeutic agents exist may attenuate the sequelae of hypoxic-ischemic cerebral injury to the term infant. PMID- 9394864 TI - Hypoxic-ischemic brain injury in the newborn. Cellular mechanisms and potential strategies for neuroprotection. AB - Recent advances have delineated many of the complex cellular mechanisms of cerebral hypoxic-ischemic injury. These developments have created opportunities for the design of rational "mechanism-based" strategies that target specific injurious processes. A number of neuroprotective agents have entered adult clinical trial and practice. For the newborn infant, progress in this areas has been judiciously delayed by toxicity concerns. Central to those safety concerns is the close relationship between mechanisms of hypoxic-ischemic cellular injury and normal developmental processes. These cellular mechanisms and the dilemmas facing the advance of this field are discussed. The likelihood of an effective single "magic bullet" neuroprotective strategy emerging in the near future appears remote. Rather more likely is the development of "cocktail" therapies that seek to exploit synergistic antagonism at multiple levels in the complex concentration of cellular events mediating hypoxic-ischemic cellular injury. However, such combination therapies will require elucidation of the complex inter relationships between mechanisms of cellular injury, brain development, and the combination of therapies envisioned. PMID- 9394866 TI - Recent advances in neonatal cranial ultrasound and Doppler techniques. AB - Use of new scanning approaches, such as the mastoid fontanelle and new signal processing techniques, such as power Doppler and spectral Doppler during fontanelle compression, have dramatically improved our ability to image both the structure and hemodynamics of blood flow in the neonatal brain. Continuing development of US contrast agents hold promise for creation of regional cerebral blood-flow maps at the cribside in critically ill newborns. PMID- 9394865 TI - Cerebrovascular complications and neurodevelopmental sequelae of neonatal ECMO. AB - A total of 355 infants have been treated with ECMO at our hospital between 1985 and 1996, 271 of whom have been enrolled in an ongoing prospective study; of the 271 infants enrolled, 223 (82%) survived, and most function within the normal range of development. Nevertheless, handicapping sequelae, including spastic forms of CP, hearing loss, and cognitive deficiencies at school age, have been noted in a significant minority of ECMO-treated survivors. The need for RCCA cannulation during venoarterial ECMO may increase the risk of a cerebrovascular injury, and lateralized CBF abnormalities have been noted on CDI and pulsed Doppler ultrasound studies during and after venoarterial bypass; however, post ECMO CT scans, HUS, MR images, or clinical evaluations have not indicated selective or greater injury to the right, compared with the left, cerebral hemisphere in our survivors, nor was there a significant predilection for right, rather than left, cerebral hemispheric EEG abnormalities during or following venoarterial bypass. Although we routinely repair the RCCA following venoarterial ECMO, the long-term consequences of a permanently ligated artery have not as yet been demonstrated. We have noted the ominous predictive value of two or more recordings that disclose ES and BS EEG abnormalities before or during venoarterial ECMO and found that the need for vigorous CPR before or during RCCA cannulation significantly increased the risk of these two markedly abnormal bioelectric patterns. Because 85% of infants with severe respiratory failure have moderate to marked EEG abnormalities (including 23% who have BS or ES patterns) before ECMO, we believe that fetal and neonatal complications related to the occurrence and treatment of severe cardiorespiratory failure are responsible in large part for the neurologic sequelae in ECMO survivors. The risk for CP was significantly increased in survivors of neonatal venoarterial ECMO treated at our hospital who required CPR or who independently had a systolic BP below 39 mm Hg before or during ECMO. We also noted that the risk for hearing loss was increased significantly in surviving neonates who had a PaCO2 below 14 mm Hg before ECMO. The possibility that undetected confounding variables were, in part, responsible for the neurologic, audiologic, and cognitive sequelae in ECMO survivors could not be excluded entirely by our data analyses. Although the pathogenesis of severe brain damage has not been defined fully in neonates treated with ECMO, focal, multifocal, or diffuse cerebral ischemia is the most likely final common pathway; thrombosis, infarction, or hemorrhage may follow and contribute to the brain injury. The cause of isolated SNHL is unknown in most affected ECMO survivors, but in some very likely is associated with the complications and treatment of severe cardiorespiratory failure, including profound hypocarbia prior to ECMO. The results of our studies to date are consistent with the following conclusions: (1) hypotension before or during ECMO and the need for CPR before ECMO contribute to the pathogenesis of CP, probably through the mechanism of cerebral ischemia; (2) profound hypocarbia before ECMO and delayed ECMO treatment are associated with a significantly increased risk of hearing loss; (3) hypoxemia without hypotension does not result in CP; (4) the type and severity of neurologic and cognitive sequelae in ECMO survivors depends, in part, on the primary cause of the neonatal cardiorespiratory failure; (5) early neurodevelopment, except for severe deficits, may not predict school-age performance; and (6) abnormally low or borderline WPPSI-R IQ scores and academic deficiencies at early school age, without evidence of a congenital abnormality of brain or CP or SNHL, remain unexplained. The criteria for initiating ECMO in the neonate with severe cardiorespiratory failure include decreasing oxygenation despite mechanical hyperventilation with 100% oxygen. (ABSTRACT TRUNCATED) PMID- 9394867 TI - Magnetic resonance techniques in the evaluation of the newborn brain. AB - MR imaging provides unequaled sensitivity as compared with US or CT scanning for evaluating developmental changes and pathologic processes in the newborn brain. Myelination can be assessed qualitatively and quantitatively using newer 3D-MR imaging methods. MR imaging provides a much clearer delineation of many developmental disorders, including anomalies of migration and organization, as well as a variety of metabolic disorders and congenital infections. Neonatal intracranial hemorrhage is detected in all its locations by MR imaging. The timing of the hemorrhage is a unique feature of MR imaging. Venous thrombosis also can be identified by MR imaging and confirmed with MR angiography. HIE is the major cause of potentially preventable or reversible brain injury that results in considerable long-term neurologic morbidity. Early detection is crucial for interventions aimed at preventing or reversing ongoing injury. DWI can show early changes at the cellular level that are not detectable by any other imaging modality. MR spectroscopy has further opened the possibility of studying the metabolic mechanisms that define the pathophysiologic events taking place in neonatal brain injury. Both 31P-MR spectroscopy, as a marker of the acute changes in energy metabolism, and 1H-MR spectroscopy, with the measurement of lactate and the excitotoxic aminoacids glutamate and glutamine, have enabled us to study the early and late effects of insults to the newborn brain in a noninvasive fashion. Studies performed to determine the predictive value of MR spectroscopy for later neurodevelopmental outcome after HIE have shown promising results but need further evaluation on larger patient samples. The potential use of these methods in the evaluation of early neuroprotective treatment regimens in the newborn remains to be determined. PMID- 9394868 TI - Radiographic patterns of pulmonary disease. AB - Pulmonary radiographs are essential adjuncts to the evaluation and diagnosis of suspected pulmonary disease. In the intensive care unit, radiographs are useful to confirm correct positioning of diagnostic and therapeutic devices. Patterns seen on the radiograph may be within broadly normal limits or may be interpreted as abnormal, especially when placed in the clinical context of a specific patient's problem. The description abnormal can be related to both nonspecific and specific radiographic patterns of disease. Nonspecific radiographic patterns of disease include location of disease, temporal course of disease, pleural abnormalities, hyperinflation, extra-alveolar air, atelectasis, bronchiectasis, and vascular disease. Specific radiographic patterns of disease are discrete anatomic structures seen on a radiograph, for example, cavitary and cystic disease. The interpretation of nonspecific and specific radiographic patterns is useful in diagnosis, selection of treatment, and monitoring of the course of disease and the patient's response to treatment. PMID- 9394869 TI - CRF/NPY interactions: a potential role in sleep dysregulation in depression and anxiety. AB - Neuropeptide Y (NPY) has neuromodulatory actions on multiple brain functions including endocrine, behavioral, and circadian processes and has been implicated in the pathophysiology of both anxiety and depression. Behavioral studies suggest that NPY is a potent anxiolytic, whereas CRF is anxiogenic, thus it seems that a balance of these two peptides may exert important influences on behavioral state regulation. However, little is known about how the NPY/CRF balance affects general arousal, attention, and/or sleep states. The present study evaluated the effects of CRF alone, and co-administered with NPY, on spontaneous brain activity as well as on auditory processing using electrophysiological measures. Electroencephalographic (EEG) and event-related potentials (ERPs) were obtained in rats following intracerebroventricular administration of CRF (0.5 microgram) and CRF (0.5 microgram)/NPY (5.0 or 15 micrograms). Auditory processing, as assessed by ERPs, was affected most significantly in the frontal cortex where CRF produced increases in the N1 and P3 components of the ERP, and NPY/CRF co administration produced significant decreases. These data are consistent with a role for CRF in hyperarousal, and further suggest that NPY may be capable of reversing such states. Administration of CRF also produced a significant increase in the time to sleep onset and a decrease in the amount of time spent in non rapid eye movement (NREM) sleep as quantified by scoring the EEG paper records. Co-administration of NPY with CRF reversed the effects of CRF on sleep duration and sleep onset in a dose-dependent fashion. Spectral analysis revealed that CRF produced quantitative changes in the EEG that were similar to what has previously been reported. CRF-induced increases in fast frequency activity were found to be reversed by co-administration of NPY. Taken together these data suggest that "dysregulation" of sleep and arousal states in depression and anxiety may be consistent with an upset of the balance between hypothalamic neuropeptide systems. PMID- 9394870 TI - Are SSRIs better than TCAs? Comparison of SSRIs and TCAs: a meta-analysis. AB - In this analysis we examined studies of tricyclic antidepressants (TCAs) and selective serotonin reuptake inhibitors (SSRIs) to compare efficacy and drop-out rates. Frequency of reported side effects was also studied. Using Medline, we located 36 clinical trials of TCAs and SSRIs in a double-blind comparison. We performed a meta-analysis on these studies and on a subgroup of 21 studies that had more uniformly defined outcome criteria. The main outcome measures were efficacy for treatment completers and for the intention-to-treat group; drop-out rates due to adverse reactions and lack of efficacy; and reported side effects. Overall, the response rate to treatment for patients who completed a trial was 63.2% for SSRIs and 68.2% for TCAs (P = 0.038). For the intention-to-treat groups, these rates dropped to 48.0 and 48.6% (P, NS), respectively. Significantly more TCA-treated than SSRI-treated subjects dropped out due to either lack of efficacy or adverse reactions (30.0 vs. 24.7%, P = 0.01). Patients taking SSRIs experienced significantly more gastrointestinal problems and sexual dysfunction, whereas treatment with TCAs produced significantly more complaints of sedation, dizziness, and anticholinergic symptoms. PMID- 9394871 TI - Dexamethasone suppression test identifies a subset of elderly depressed patients with reduced platelet serotonin transport and resistance to imipramine inhibition of transport. AB - Dysregulation of the hypothalamus-pituitary-adrenal axis (HPA) is more common in elderly patients with depression than in younger depressed patients, and glucocorticoids are known to influence serotonergic function. Elderly depressed patients are also reportedly more resistant to therapeutic effects of antidepressants. In the current study, we measured platelet serotonin transporter binding sites and transport function in young and elderly depressed patients and determined the relationship to HPA status as assessed with the dexamethasone suppression test (DST). The density and affinity of transporter molecules showed no differences between young and elderly depressed patients, regardless of DST results. Nevertheless, transporter function showed a substantial interaction of aging with DST: elderly DST suppressors showed a deficit in [3H]serotonin uptake capabilities and resistance to imipramine inhibition of uptake. No such defects were seen in the young depressed cohort, regardless of DST status, nor in elderly depressed DST non-suppressors. These results are consistent with the view that depression in the elderly exhibits basic biological differences from depression in earlier life, and that such distinctions may account in part for therapeutic ineffectiveness of antidepressants in specific subgroups, associated with the presence or absence of appropriate HPA regulation. PMID- 9394872 TI - Effect size of efficacy measures comparing divalproex, lithium and placebo in acute mania. AB - Effect size (ES) is a statistical concept that can be used to improve the interpretation of results from psychopharmacological studies. ES may aid interpretation of results when sample size is unbalanced or small or when units or levels of baseline measures differ across items. Usually, an investigator can define a threshold value for a clinically meaningful ES based on published data and clinical judgment or by resorting to conventions, e.g., a medium ES = 0.5 S.D., which can usually be discerned by the trained clinician. In the present study, we apply ES analysis to results from a study comparing the effectiveness of divalproex (DIVAL), lithium (LI), and placebo (PLA) in hospitalized, acutely manic patients. One hundred seventy-six patients were randomly assigned to DIVAL, LI, or PLA in a 2:1:2 ratio, with drug administered in a double-blind, parallel group design for 21 days. The primary efficacy measure was the Mania Rating Scale from the Schedule for Affective Disorders and Schizophrenia, composed of the Manic Syndrome Score (MSS) from items that are relatively specific to the manic state, and the Behavior and Ideation Score (BIS), which reflects severe but nonspecific psychopathology. Improvement of the MSS after 5 days of treatment was difficult to interpret based on percentage change (DIVAL = 19%, LI = 13.5%, PLA = 8.5%). However, the corresponding effect sizes of 0.79, 0.55, and 0.35 indicated a medium to marked ES for DIVAL, a medium ES for LI, and a small ES for PLA at this early point in treatment. Similarly, the ES for change on the MSS at the end of treatment indicated a large, readily observable improvement with both DIVAL (ES = 1.01) and LI (ES = 0.79) vs. an ES of 0.37 for PLA. ES analysis also indicated that the BIS is a less robust indicator of change to either drug. The ES at the end of treatment for the BIS was 0.67 for DIVAL-, 0.62 for LI-, and 0.25 for PLA-treated patients. PMID- 9394873 TI - Fluoxetine and concomitant centrally acting medication use during clinical trials of depression: the absence of an effect related to agitation and suicidal behavior. AB - Concomitant use of psychoactive medications is a common practice in most clinical trials of antidepressant medications. However, the relative therapeutic impact of such use on trial results has not been the subject of much attention. We conducted a meta-analysis to determine whether concomitant use of psychoactive medications confounded the efficacy or safety results of a series of fluoxetine trials. Data were evaluated from 25 randomized, double-blind clinical trials comparing fluoxetine with placebo or a tricyclic antidepressant (TCA) in 4,016 patients with major depression. We compared incidence rates of concomitant use of anxiolytics, sedatives, and antipsychotics between treatments. In addition, we compared the change in total score for the 21-Item Hamilton Depression Rating Scale (HAMD21): incidence rates of any worsening, emergence, or improvement in psychomotor agitation; and incidence of suicidal acts and any worsening, emergence, or improvement in suicidal ideation between treatment groups among patients taking/not taking a sedative. Anxiolytic and antipsychotic drug use was uncommon (8.3% and 0.9% overall use, respectively) and did not substantially increase over time. Sedative drugs were used most often (29.6% overall), but only 29.8% of the fluoxetine-treated patients took one or more doses. Regarding efficacy, fluoxetine was superior to placebo in decreasing HAMD21 total scores among patients taking/not taking sedatives. Effects on safety were assessed by examining agitation and suicidal ideation. Use of sedatives did not affect the change in the HAMD agitation score; scores were similar in patients receiving fluoxetine, placebo, and TCAs. In all treatment groups, anxiolytic use tended to increase as the HAMD anxiety score increased. Fluoxetine was superior to placebo in treating suicidal ideation, and the concomitant use of sedatives did not influence this effect. Overall, concomitant use of psychotropic medications in the fluoxetine depression clinical trials was uncommon. Our meta-analysis demonstrated that the clinical efficacy and safety of fluoxetine were not confounded by the concomitant use of medications. PMID- 9394874 TI - Psychotic versus nonpsychotic depression in hospitalized adolescents. AB - One hundred fifty adolescent inpatients with major depression were systematically assessed for demographic and clinical differences between psychotic and nonpsychotic depression. Delusions and/or hallucinations were present in 10% of the subjects. The psychotic group had significantly more frequent and severe suicidal ideation. Posttraumatic stress disorder was also more frequent in the psychotic group. PMID- 9394877 TI - The issue of extralabel drug use. PMID- 9394878 TI - Help wanted: ultrasonographers. PMID- 9394879 TI - Peer review of case management skills in clinical veterinary medicine--is it time? PMID- 9394880 TI - Clinical relevance of intestinal reperfusion injury in horses. PMID- 9394881 TI - What is your diagnosis? Avulsion fracture of the proximal suspensory ligament and the third metacarpus. PMID- 9394882 TI - Myocardial necrosis and sudden death after an episode of aggressive behavior in a dog. PMID- 9394883 TI - Animal behavior case of the month. PMID- 9394884 TI - Multimedia case-simulation computer program for teaching veterinary nutrition. PMID- 9394885 TI - Communicating with clients--informed consent. PMID- 9394886 TI - How important is intestinal reperfusion injury in horses? PMID- 9394887 TI - Concentration of ionized calcium in plasma from cats with urethral obstruction. AB - OBJECTIVE: To measure ionized calcium concentration in plasma from cats with urethral obstruction and to correlate these values with results of clinical biochemical analyses and physical examinations. DESIGN: Prospective study. ANIMALS: 24 male cats. PROCEDURE: Blood samples were obtained from each cat on admission, and PCV, pH, and concentrations of ionized calcium, total calcium, glucose, total solids, sodium, potassium, BUN, creatinine, chloride, magnesium, albumin, and phosphorus were determined. Mentation, tissue perfusion, and ECG recordings were also assessed. RESULTS: 18 (75%) cats had low ionized calcium concentrations (reference range, 2.4 to 2.8 mEq/L). Hypocalcemia was considered mild (2.0 to 2.36 mEq/L) in 9 (37.5%) cats, moderate (1.6 to 1.98 mEq/L) in 6 (25%), and severe (< 1.6 mEq/L) in 3 (12.5%). Significant positive correlations were found between ionized calcium concentration and heart rate, pH, and concentrations of sodium, chloride, and total calcium. Significant negative correlations were found between ionized calcium concentration and concentrations of potassium, BUN, creatinine, and phosphorus. CLINICAL IMPLICATIONS: Most cats with urethral obstruction had a low concentration of ionized calcium. This may contribute to cardiac electrical and mechanical dysfunction in some severely affected cats. Although effects of i.v. administration of calcium were not evaluated, results of this study strengthen the rationale for its use in cats with urethral obstruction. PMID- 9394888 TI - Measurement of serum total thyroxine, triiodothyronine, free thyroxine, and thyrotropin concentrations for diagnosis of hypothyroidism in dogs. AB - OBJECTIVE: To determine whether measurement of baseline serum concentrations of total thyroxine (T4), and triiodothyronine (T3), free T4, and thyrotropin (thyroid-stimulating hormone; TSH) would aid in the diagnosis of hypothyroidism in dogs. DESIGN: Prospective case series. ANIMALS: 54 dogs with hypothyroidism, 54 euthyroid dogs with nonthyroidal disease initially suspected to have hypothyroidism, and 150 clinically normal dogs. PROCEDURE: In the 54 dogs with hypothyroidism, diagnosis was established on the basis of clinical signs, results of routine laboratory and TSH stimulation tests, exclusion of concurrent nonthyroidal disease, and a good clinical response to treatment with L-thyroxine. Blood samples were collected from all dogs and were tested for thyroid hormone and TSH concentrations. Reference ranges for hormone concentrations were established on the basis of results for the 150 clinically normal dogs. RESULTS: Of the 54 hypothyroid dogs, 48 (89%) had low total T4 concentrations, 3 had low normal concentrations, and 3 had high concentrations because of T4 autoantibodies. In contrast, only 10 (18%) euthyroid dogs had low total T4 concentrations. Only 3 of 31 (10%) hypothyroid dogs had low T3 concentrations; 23 had concentrations within the reference range, and 5 had high concentrations because of T3 autoantibodies. Only 3 of 38 euthyroid dogs had low T3 concentrations. Of the hypothyroid dogs, 53 (98%) had low free T4 concentrations and 1 had a low-normal concentration. Only 4 (7%) euthyroid dogs had low free T4 concentrations. Of the hypothyroid dogs, 41 (76%) had high TSH concentrations, and 13 had TSH concentrations within the reference range. Of the euthyroid dogs, only 4 (8%) had high TSH concentrations. Of all single hormone measurements evaluated, measurement of free T4 concentration had the highest sensitivity (0.98), specificity (0.93), and accuracy (0.95) as a test for hypothyroidism; measurement of total T4 concentration had a lower sensitivity (0.89), specificity (0.82), and accuracy (0.85). Compared with measurement of total or free T4 concentration, measurement of TSH concentration had a lower sensitivity (0.76) and accuracy (0.84) but specificity (0.93) equal to that for measurement of free T4 concentration. When T4 (total or free) and TSH concentrations were evaluated together, specificity was higher than when T4 or TSH concentration was evaluated alone. Only 1 euthyroid dog had low T4 (total and free) and high TSH concentrations. CLINICAL IMPLICATIONS: Results indicate that measurement of serum free T4 and TSH concentrations is useful for diagnosis of hypothyroidism in dogs. About a quarter of the dogs with confirmed hypothyroidism, however, will have serum TSH concentrations within reference limits. PMID- 9394889 TI - Regional anesthesia of the infraorbital and inferior alveolar nerves during noninvasive tooth pulp stimulation in halothane-anesthetized dogs. AB - OBJECTIVE: To document that regional anesthesia of the infraorbital and inferior alveolar nerves would abolish reflex-evoked muscle action potentials (REMP) in the digastricus muscle during noninvasive stimulation of tooth pulp in halothane anesthetized dogs. DESIGN: Prospective study. ANIMALS: 9 healthy female dogs between 2 and 6 years old. PROCEDURE: Dogs were anesthetized using halothane. An alligator clip anodal electrode was attached to the tooth to be stimulated, and a platinum needle cathodal electrode was inserted in adjacent gingival mucosa. The cathodal and anodal electrodes were moved to the left upper and lower canine, fourth premolar, and first molar teeth for sequential stimulation. Baseline recording of REMP was made for each tooth. Catheters were inserted percutaneously in the infraorbital and mandibular canals. Saline (0.9% NaCl) solution was injected at each catheterized site in 3 control dogs, and chloroprocaine hydrochloride was injected at each catheterized site in 6 test dogs. Each tooth was stimulated every 10 minutes for 90 minutes (test dogs) or every 10 minutes for 30 minutes and at 90 minutes (control dogs), and REMP was recorded. RESULTS: REMP was abolished within 10 minutes in all test dogs, except during stimulation of the lower first molar in 1 dog. In 4 dogs, duration of blockade was less than 90 minutes. The REMP was not restored within 90 minutes for the upper teeth in 1 dog and within 2 hours for all teeth in another dog. At 24 hours, REMP was restored for all teeth except the lower left canine in 1 dog. The REMP was restored for the lower left canine in that dog at 96 hours. The REMP was not abolished at any time in control dogs. CLINICAL IMPLICATIONS: Regional anesthesia of the infraorbital and inferior alveolar nerves may effectively provide analgesia for dental procedures in dogs. PMID- 9394890 TI - Risk factors for acquired megaesophagus in dogs. AB - OBJECTIVE: To identify risk factors associated with acquired megaesophagus in dogs. DESIGN: Case-control study. ANIMALS: 136 dogs with acquired megaesophagus (case dogs); 272 dogs from the general hospital population and 151 dogs that underwent thyroid-stimulating hormone response tests (control dogs). All dogs were more than 6 months old. PROCEDURE: Medical records of dogs in which megaesophagus was diagnosed during a 10-year period were reviewed. Inclusion criteria included regurgitation or vomiting, onset of clinical signs at more than 6 months of age, and radiographic evidence of generalized esophageal dilatation. Dogs with intra- or extraesophageal obstructive disease, brain stem disease, or neck trauma were excluded from analyses. Statistical analyses included odds ratios, 95% confidence intervals, and two-tailed t-tests. Control dogs were frequency matched to case dogs on the basis of year of diagnosis. RESULTS: Dogs with megaesophagus ranged from 0.75 to 18 years old (mean, 8.1 years) and were significantly older and heavier than control dogs. More males than females were affected, but sex and reproductive status were not associated with megaesophagus. German Shepherd Dogs, Golden Retrievers, and Irish Setters were at increased risk for developing megaesophagus. Peripheral neuropathies, laryngeal paralysis, acquired myasthenia gravis, esophagitis, and chronic or recurrent gastric dilatation with or without volvulus were associated with an increased risk of developing megaesophagus. Hypothyroidism was not associated with megaesophagus. CLINICAL IMPLICATIONS: Dogs with acquired megaesophagus should be evaluated for peripheral neuropathies, laryngeal paralysis, acquired myasthenia gravis, esophagitis, and chronic or recurrent gastric dilatation with or without volvulus. These dogs may be evaluated for hypothyroidism; however, this study did not reveal a clear association between hypothyroidism and acquired megaesophagus. PMID- 9394892 TI - Complete deletion of factor IX gene and inhibition of factor IX activity in a labrador retriever with hemophilia B. AB - Hemophilia B is a heritable bleeding disorder caused by mutations in the gene coding for coagulation factor IX. The defect has been identified in purebred and mixed-breed dogs. Management of affected dogs requires transfusion of canine blood products supplying active factor IX. Production of inhibitors to factor IX is a complication of transfusion therapy that has been documented as affecting human patients. Risk for producing coagulation inhibitors is greatest for patients having large factor IX gene deletions. To our knowledge, this is the first report of canine factor IX inhibitor production in dogs. The affected dog had clinically severe hemophilia B caused by complete deletion of the factor IX gene and developed resistance to transfusion. Comprehensive evaluation of hemophilic dogs, including assays of specific factor activity, concentration, and factor inhibition, enhances diagnosis and management of this bleeding disorder. Characterization of the molecular defect causing hemophilia is useful for genetic counseling and identifying individuals at highest risk for producing coagulation inhibitors. PMID- 9394891 TI - Multicenter clinical comparison of sedative and analgesic effects of medetomidine and xylazine in dogs. AB - OBJECTIVE: To evaluate analgesic and sedative effects of medetomidine hydrochloride in dogs and to compare effects with those of xylazine hydrochloride. DESIGN: Randomized, controlled trial. ANIMALS: 184 dogs that required sedation or analgesia for completion of minor diagnostic or therapeutic procedures. PROCEDURE: Dogs were sedated with medetomidine, i.v. (750 micrograms/m2 of body surface area) or i.m. (1,000 micrograms/m2) or with xylazine, i.v. (1.1 mg/kg 10.5 mg/lb] of body weight) or i.m. (2.2 mg/kg [1 mg/lb]). Sedative effects were measured by scoring posture and response to noise. Durations of effects were determined by measuring time intervals between drug administration and changes in posture. Analgesic effects were measured by determining toe-pinch pressure needed to elicit a withdrawal response. Clinicians rated sedative and analgesic effects and ease with which diagnostic or therapeutic procedures could be performed. RESULTS: Posture and response to noise scores were significantly higher for dogs given medetomidine, i.m., than for dogs given xylazine, i.m., and for dogs given medetomidine, i.v., than for dogs given xylazine, i.v. Time to regaining sternal recumbency and time to regaining ability to stand were longest after i.m. administration of medetomidine. Toe-pinch pressures were not significantly different among groups. Clinicians rated overall analgesic and sedative effects as excellent significantly more often after administration of medetomidine than after administration of xylazine. Prevalence of adverse effects did not differ among groups. CLINICAL IMPLICATIONS: Medetomidine and xylazine, at doses tested, were effective and safe, but results of subjective measurements indicated that medetomidine provided better sedation and analgesia than did xylazine. Specific alpha 2-adrenergic antagonists (atipamezole, yohimbine) are available for control of adverse cardiovascular effects. PMID- 9394893 TI - Evaluation of prognostic factors for dogs with primary lung tumors: 67 cases (1985-1992). AB - OBJECTIVE: To determine associations between clinical and histologic factors in dogs with primary lung tumors and outcome and to develop a histologic grading method for primary lung tumors. DESIGN: Retrospective study. ANIMALS: 67 dogs undergoing thoracotomy and lobectomy for primary lung tumors. PROCEDURE: Medical records and histologic sections were reviewed to evaluate factors of prognostic importance. Association of these factors with disease-free interval (DFI) and survival time was evaluated, using the Cox proportional hazards model. Median DFI and survival time were determined, using the Kaplan-Meier product-limit method. RESULTS: Clinical and histologic factors significantly associated with prognosis were histologic score, detection of clinical signs, and metastasis to regional lymph nodes. On the basis of histologic score, a histologic grading method was developed. Dogs with well-differentiated tumors had significantly longer survival time and DFI (median DFI, 493 days) than dogs with moderately (median DFI, 191 days) or poorly (median DFI, 0 days) differentiated tumors. Dogs with clinical signs or metastasis to regional lymph nodes had shorter survival times and DFI than dogs in which lung masses were discovered as an incidental finding. CLINICAL IMPLICATIONS: Dogs with well-differentiated, nonmetastasized, primary lung tumors that do not have clinical signs associated with the tumor have a favorable prognosis. Dogs with more advanced disease or aggressive tumors histologically may require treatment, such as chemotherapy in combination with surgery. The grading method proposed here for primary lung tumors may be useful in other dogs with primary lung tumors. PMID- 9394894 TI - Risk factors for acquired myasthenia gravis in dogs: 1,154 cases (1991-1995). AB - OBJECTIVE: To determine frequency of initial clinical signs and risk factors for acquired myasthenia gravis (MG) in dogs. DESIGN: Retrospective study. SAMPLE POPULATION: 1,154 dogs residing within the United States from 1991 to 1995 with a confirmed diagnosis of acquired MG and 7,176 dogs with other neuromuscular disorders, including generalized weakness, megaesophagus, and dysphagia (control group). PROCEDURE: Records were retrieved from a database containing results of serum samples tested for acetylcholine receptor antibodies. Signalment, breed, age, state of origin, and month of onset of clinical signs were obtained. An antibody titer > 0.6 nmol/L was diagnostic for acquired MG. Unconditional logistic regression was used for statistical analysis. RESULTS: In comparison with mixed-breed dogs, dogs with the highest risk of acquired MG were Akitas, terrier group, Scottish Terriers, German Shorthaired Pointers, and Chihuahuas. Rottweilers, Doberman Pinschers, Dalmatians, and Jack Russell Terriers had low relative risks. Sexually intact males and dogs less than 1 year old had some protection from risk. Generalized weakness with megaesophagus and megaesophagus alone were the most common initial clinical signs. CLINICAL IMPLICATIONS: Breed predispositions for acquired MG were demonstrated. Age and sex were contributing factors. Although most dogs had generalized clinical signs, a substantial proportion of dogs had focal signs. PMID- 9394895 TI - Renal transplants in cats: 66 cases (1987-1996). AB - OBJECTIVE: To document the morbidity and survival time after renal transplants in cats with end-stage renal failure. DESIGN: Retrospective case series. ANIMALS: 66 cats that had renal transplants. PROCEDURE: Information regarding signalment, history, diagnostic testing, and postoperative morbidity and mortality was retrieved from medical records of cats with renal failure that had renal transplants at the University of California School of Veterinary Medicine between 1987 and 1996. RESULTS: 47 of 66 (71%) cats that had renal transplants survived until discharge. Nineteen cats died in the perioperative period. Most common causes of death were seizure-related complications (7 cats) and renal pedicle complications (4). One discharged cat was unavailable for follow-up monitoring. Of the 46 cats discharged and available for follow-up monitoring, 28 died. Most common causes of death in these cats were renal complications (9 cats) and death related to immunosuppression (8; mean and median survival times, 15 and 12 months, respectively). Of the 18 cats that were still living at the time this report was written, mean and median survival times were 26 and 22 months, respectively. CLINICAL IMPLICATIONS: Renal transplantation resulted in long-term survival of many cats that would have otherwise died from, or have been euthanatized as a result of, renal failure. Problems with ureteral obstruction can be minimized. Postoperative CNS disorders were the most prevalent complication. PMID- 9394896 TI - Mechanisms of intestinal mucosal repair. PMID- 9394897 TI - Respiratory function parameters in infants using inductive plethysmography. AB - A signal processing technique has been developed to determine respiratory function parameters by processing displacement data obtained from the abdomen and the rib cage of infants using respiratory inductive plethysmography. The technique transforms time-variant signals into the frequency domain, where they are filtered to reduce unwanted signal components. The phase relationship between the abdominal displacement and the rib cage displacement is determined from the phases of the filtered signals. Flow-volume loops, which are currently of great interest in respiratory medicine, are obtained from waveforms representing respiratory tidal flow and respiratory tidal volume. The index of respiratory timing is determined from a waveform representing respiratory tidal flow. The technique has been verified by statistical comparison with data simultaneously obtained using pneumotachography in a clinical study involving 49 infants. PMID- 9394898 TI - Estimating the effective Young's modulus of soft tissues from indentation tests- nonlinear finite element analysis of effects of friction and large deformation. AB - A nonlinear finite element model was developed to investigate the biomechanics of indentation, particularly the influence of friction and large deformation on the calculation of the effective Young's modulus from the cylindrical, flat-ended indentation test of soft tissues. A new kappa table was given for calculation of the effective Young's modulus to account for the effects of layered geometry with consideration of the larger deformation. The results indicate that the effect of friction on the calculation of Young's modulus becomes significant with a large aspect ratio and with a large Poisson's ratio. It is found that the factor kappa increases almost proportionally to the increase of the indentation depth, especially obvious with a larger Poisson's ratio v and a larger aspect ratio a/h. PMID- 9394899 TI - A stiffness-varying model of human gait. AB - We report on a conceptual two degrees of freedom (2 DOF) human gait model, which incorporates nonlinear joint stiffness as a stabilizing agent. Specifically, muscle spring-like property provides inherent stability during gait movement using a nonlinear angular spring and dash pot at each joint. The instability problem of the gait model in direct dynamic analysis is overcome by simulating the human co-contraction muscle function. By developing dynamic system stability requirements and hypothesizing a minimum joint stiffness criterion, we determine time-varying joint stiffness. Optimum joint stiffnesses are present for varying gait pattern, stride lengths and cadences. We conclude that nonlinear joint stiffness can be incorporated into gait models to overcome stability problems inherent in such linkage models. PMID- 9394901 TI - Modelling and simulation of the hand grasping using neural networks. AB - In this paper we present preliminary results of a study on the use of artificial neural networks to model and simulate the hand grasping. Results of this study will provide a basic understanding of the co-ordination and control of multiple degrees of freedom upper limb prosthetic devices and robotic end effectors when interacting with the environment. We assumed the hand to be a black box with the inputs being the object and simulation time sequence, whilst the output is the grasping postures over time. We trained the network with samples of key postures of the hand grasping several object shapes and sizes. The back-propagation technique was used to update the weights of the network. We found that the neural network is able to reproduce the postures of the hand grasping objects of different shapes and sizes from a single set of neural network weights. PMID- 9394900 TI - Apparatus and methods for studying artificial feedback-control of the plantarflexors in paraplegics without interference from the brain. AB - Apparatus has been built to explore the practical feasibility of using automatic control with electrical stimulation of paralysed legs to restore function. The experiments are performed with paraplegics with the aim of achieving a realistic postural task: to see whether the body may be maintained upright by stimulation of the plantarflexors when the other joints are braced. Significantly, the intact upper body, under natural control of the brain, cannot interfere with the automatic control. The "Wobbler" apparatus allows measurement of the ankle muscle properties in isometric conditions or in sinusoidal motion. Using the biomechanical properties of the body, which are also measured, controllers for stabilising the body can be designed. Controllers can be dynamically tested, imitating anterior-posterior sway, while the body is held upright, before "actual standing" is attempted. PMID- 9394902 TI - Comparison of loads on internal spinal fixation devices measured in vitro and in vivo. AB - The loads on internal spinal fixation devices were measured using modified, telemeterized AO-Dick internal fixators. The implants allow measurement of the three force components and the three moments acting on the implant. The modified fixators were mounted on cadaver spines, and the implant loads were measured in the intact and postcorpectomy spines for different loading modes, including axial compression force, flexion, extension, lateral bending, and torsion. The in vitro experiment did not consider muscle forces. Modified fixators were also implanted in three patients, and the implant loads were determined before and after anterior interbody fusion with autologous iliac-crest bone grafts. The results for different in vitro loading modes were compared with those in vivo in order to demonstrate the extent to which the in vitro loads represent the real situation in patients. In several cases, the implant loads in the in vitro experiment differed strongly from those measured in patients. For flexion and lateral bending, a tensile axial force occasionally was measured in the in vitro experiment, while in the patients the axial force was always compressive. Extension was predominantly associated with extension bending moments in the in vitro study but with flexion bending moments in the patients. When muscle forces are not considered in the in vitro experiment, the loads on the fixators may differ significantly from the situation found in patients. PMID- 9394903 TI - A model study of periodic breathing, stability of the neonatal respiratory system, and causes of sudden infant death syndrome. AB - A mathematical model of the neonatal respiratory system has been modified and used to examine the system under various physiological conditions at different stages of maturity. The respiratory responses in hypoxia, periodic breathing and following a sign have been analyzed. The effects of different respiratory parameters on the stability of the system for normal and premature infants have been investigated. The causes of periodic breathing, apnea spells and sudden infant death syndrome for full-term and premature infants have been studied, and the results compared with the available experimental findings. The response of the infant respiratory system has been found to be highly sensitive to several parameters of the system, as indicated by the results of this study. These significant parameters are sensitivity factor of central receptors to carbon dioxide, sensitivity factor of arterial receptors to carbon dioxide, sensitivity factor of arterial receptors to oxygen, functional residual capacity of the lungs, the alveolar-arterial oxygen difference and the lungs shunt ratio. It has been shown that different parts of the respiratory controller have antagonistic effects on hypoxic periodic breathing and apnea of infancy. PMID- 9394904 TI - Predicting the leakage performance of small bodyworn disposable incontinence pads using laboratory tests. AB - An international multi-centre project has been run to create an international standard for measuring the leakage performance of small, disposable incontinence pads for lightly incontinent women. One hundred and thirteen women tested batches of nine different incontinence pads of widely differing designs and noted the severity with which each individual used pad had leaked so that leakage performance could be determined as a function of urine weight. In addition, testers rated the overall leakage performance of each of the nine products on a five-point scale. These clinical data were compared with laboratory data from 153 different pad measurements, each of which was evaluated by seeing how well the data it yielded correlated with the clinical test data. A wetback test emerged as the clear winner. It usually predicted the clinical leakage performance of pads to an accuracy of +/- 10%. It involved applying 25 ml of 1% w/v saline to a pad and measuring how much escaped into a filter paper held against the wet pad for 1 min under a pressure of 1.5 kPa. Pads which released the least test fluid into the filter paper leaked least in the user tests. The method will be published as an ISO standard during 1997. PMID- 9394905 TI - Data compression of fetal Doppler ultrasound audio signals using zero-crossings analysis. AB - This paper outlines a method of reducing the data rate for transmitting fetal Doppler ultrasound audio signals. A specific application is cited where compression of the fetal Doppler signal is required to transmit information to cardiotocographs (CTGs) using radio telemetry. The method involves splitting the signal into amplitude and frequency components. The amplitude is represented by samples of the signal envelope whilst the frequency information is represented by the number of zero-crossings within fixed intervals (windows). With a careful choice of window size, it is shown that this method can be used to reproduce the signal with no audible difference when compared with the original waveform. A reduction in data rate of 15:1 is achieved. PMID- 9394906 TI - Time-frequency distribution of heart rate variability below 0.05 Hz by Wigner Ville spectral analysis in congestive heart failure patients. AB - The Wigner-Ville spectral analysis was utilized to demonstrate a high-resolution time-frequency distribution of heart rate variability below 0.05 Hz. There are different time-frequency characteristics between the normal subject and the patient with severe congestive heart failure. The former consists of multiple and broad-band spectral peaks, while the latter presents unique spectral peaks. Based on Bayes theory, a classifier for the unique spectral peaks was developed. After the beneficial improvement with low-dose beta-blockers, the unique spectral peaks had disappeared or the time of occurrence was reduced in most patients. PMID- 9394907 TI - Three-dimensional stress analysis of polypropylene leaflets for prosthetic heart valves. AB - The effect of changing the modulus and thickness of the material in the leaflets of an artificial heart valve has been investigated. This has been achieved with a finite element model of the valve having approximately 2300 thin shell elements. The valve motion and the resulting stresses are modelled dynamically during closure, and subsequent pressurisation. The stresses decrease as the leaflets are made thicker and the modulus is increased. Local and global thickening has been investigated. The highest stresses appear at the tops of the stent posts in the regions of the commissures. When the modulus is too low, or the leaflets are too thin, the valve prolapsed. PMID- 9394908 TI - High-risk superficial bladder cancer: transurethral resection alone in selected patients with T1 tumor. PMID- 9394909 TI - High-risk superficial bladder cancer: intravesical therapy for T1 G3 transitional cell carcinoma of the urinary bladder. AB - The ideal treatment for T1 G3 transitional cell carcinoma (TCC) of the urinary bladder remains controversial. Therapeutic options after the initial transurethral (TUR) resection are observation, intravesical therapy, a repeat resection, radiation therapy, and cystectomy. Because more than half of patients with T1 G3 TCC of the urinary bladder do not progress, initial cystectomy can represent overtreatment. However, observation alone following TUR for T1 G3 TCC of the urinary bladder is associated with a progression rate of 48%. Intravesical immunotherapy has been shown to decrease recurrence and progression in high-grade Ta carcinoma in situ and T1 bladder cancer. When patients with T1 G3 tumors are well selected, intravesical therapy following the initial TUR can significantly improve survival and quality of life. Persistence or recurrence of high-grade tumor mandates consideration of cystectomy. PMID- 9394910 TI - Early cystectomy for clinical stage T1 transitional cell carcinoma of the bladder. AB - Early radical cystectomy for a high-grade tumor invading the lamina propria (T1) remains controversial. In 1997, we cannot identify accurately which of these high risk tumors will progress to muscle-invasive disease and metastases. In the near future, urologists may be able to use the presence of genetic alterations, such as p53 mutations, to help make therapeutic decisions. Previous reports on superficial bladder cancer treated with intravesical bacillus Calmette-Guerin immunotherapy have demonstrated a decrease in recurrence and progression. Unfortunately, there is no reliable method to predict which patients with a high grade T1 tumor will fail to respond to intravesical therapy. Failure of intravesical therapy to control these aggressive tumors is associated with a significant rate of pathological upstaging and metastases. Radical cystectomy will cure a high percentage of these T1 tumors with acceptable morbidity and low mortality. In an era of nerve-sparing cystectomy and orthotopic neobladder reconstruction, early radical cystectomy is an alternative that should be discussed with the patient before instituting intravesical therapy. PMID- 9394911 TI - The case for radiotherapy with or without chemotherapy in high-risk superficial and muscle-invading bladder cancer. AB - The standard treatment for superficial high-grade bladder cancer is transurethral resection with or without subsequent intravesical therapy. Although a few series have reported good local control rates for T1 tumors, using either external beam irradiation or brachytherapy, this does not represent the standard of care in the United States. External beam radiation may be attempted in patients whose tumors cannot be resected transurethrally and who refuse cystectomy. The case for radiotherapy with or without chemotherapy is far stronger in muscle-invading cancers. Overall survival rates around 50% have been reported in larger series from a number of major centers. Most of these 5-year survivors retain their native bladders. The bladder morbidity of such an approach is very low. There are several studies currently active to determine the most appropriate sequence and combination of drugs and radiation. PMID- 9394912 TI - Superficial bladder cancer: the role of molecular markers in the treatment of high-risk superficial disease. AB - The use of molecular markers to guide decision making in the treatment of high risk superficial disease is in its infancy. At present, p53 expression and epidermal growth factor receptor expression have been studied in greatest depth. The abundance of potential markers and our greater understanding of cellular pathophysiology suggest that molecular markers with significant stratifying power beyond that provided by standard histology shall be identified in the near future. PMID- 9394913 TI - Continent cutaneous urinary diversion. AB - Using the case history of a 57-year-old woman with a G3 T1 bladder cancer as a reference point, general considerations on tumor biology and therapeutic choices are reviewed. Options for urinary tract diversion or reconstruction are presented. This author recommends radial cystectomy for this patient with continent cutaneous urinary diversion, the Indiana pouch. Important technical points of the surgical procedure are emphasized. PMID- 9394914 TI - Orthotopic diversion in women. AB - An orthotopic neobladder has quickly become the preferred diversion after cystectomy in most women. The decision to proceed with a cystectomy for high grade, superficially invasive transitional cell carcinoma is frequently difficult for the patient and physician alike. The availability of an orthotopic diversion to provide an excellent voiding pattern and continence may tip the scale toward earlier cystectomy with anticipated excellent cancer control. PMID- 9394915 TI - Comparison of the ileal conduit to the continent cutaneous diversion and orthotopic neobladder in patients undergoing cystectomy: a critical analysis and review of the literature. AB - In recent years, many investigators have devised innovative urinary diversions to improve the quality of life of patients requiring urinary diversions after cystectomy. In this article, we report a literature review in which we compare the outcomes of patients receiving an ileal conduit to those receiving either a continent cutaneous diversion or an orthotopic neobladder. In this discussion, we analyze the data in the literature that pertains to quality of life, incontinence rates, and metabolic complications. We conclude that although the newer urinary diversions are promising, randomized prospective trials comparing patients receiving these diversions to those receiving an ileal conduit need to be performed. PMID- 9394916 TI - Nursing issues in the management of urinary diversions in women. AB - Cancer and urinary diversion via orthotopic or cutaneous stomas in women result in significant physiological and psychosocial issues. Patient counseling, education, and follow-up care are best achieved by the intervention of a multidisciplinary health care team. Urologic and enterostomal therapy nursing support in these endeavors can contribute to facilitating a positive outcome. PMID- 9394917 TI - Role of Rab GTPases in membrane traffic. AB - Small GTPases of the Rab subfamily have been known to be key regulators of intracellular membrane traffic since the late 1980s. Today this protein group amounts to more than 40 members in mammalian cells which localize to distinct membrane compartments and exert functions in different trafficking steps on the biosynthetic and endocytic pathways. Recent studies indicate that cycles of GTP binding and hydrolysis by the Rab proteins are linked to the recruitment of specific effector molecules on cellular membranes, which in turn impact on membrane docking/fusion processes. Different Rabs may, nevertheless, have slightly different principles of action. Studies performed in yeast suggest that connections between the Rabs and the SNARE machinery play a central role in membrane docking/fusion. Further elucidation of this linkage is required in order to fully understand the functional mechanisms of Rab GTPases in membrane traffic. PMID- 9394918 TI - Lignification in plant cell walls. AB - Cell wall lignification is a complex process occurring exclusively in higher plants; its main function is to strengthen the plant vascular body. This process involves the deposition of ill-defined phenolic polymers, the so-called lignins, on the extracellular polysaccharidic matrix. These polymers arise from the oxidative coupling of three cinnamyl alcohols in a nonrandom reaction, in which cell wall polysaccharides appear to influence the freedom of cinnamyl alcohol radicals, giving rise to a highly orchestrated process. This review is focused on the most recent advances in the chemical, biochemical, cytological, physiological, and evolutive aspects of cell wall lignification. As we shall see throughout this review, there are still some open questions to be answered which may serve as the basis of future endeavors. PMID- 9394919 TI - Functional interactions among cytoskeleton, membranes, and cell wall in the pollen tube of flowering plants. AB - The pollen tube is a cellular system that plays a fundamental role during the process of fertilization in higher plants. Because it is so important, the pollen tube has been subjected to intensive studies with the aim of understanding its biology. The pollen tube represents a fascinating model for studying interactions between the internal cytoskeletal machinery, the membrane system, and the cell wall. These compartments, often studied as independent units, show several molecular interactions and can influence the structure and organization of each other. The way the cell wall is constructed, the dynamics of the endomembrane system, and functions of the cytoskeleton suggest that these compartments are a molecular "continuum," which represents a link between the extracellular environment and the pollen tube cytoplasm. Several experimental approaches have been used to understand how these interactions may translate the pollen-pistil interactions into differential processes of pollen tube growth. PMID- 9394920 TI - Biology of the postsynaptic glycine receptor. AB - Glycine is one of the major inhibitory neurotransmitters, and upon binding to its receptor it activates chloride conductances. Receptors are accumulated immediately opposite release sites, at the postsynaptic differentiations, where they form functional microdomains. This review describes recent advances in our understanding of the structure-function relationships of the glycine receptor, a member of the ligand-gated ion channel superfamily. Following purification of the receptor complex and identification of its integral and peripheral membrane protein components, molecular cloning has revealed the existence of several subtypes of the ligand-binding subunit. This heterogeneity is responsible for the distinct pharmacological and functional properties displayed by the various receptor configurations that are differentially expressed and assembled during development. This review also focuses on the molecular aspects of glycinergic synaptogenesis, highlighting gephyrin, the peripheral component of the receptor. The role of this cytoplasmic protein in anchoring and maintaining the channel complex in postsynaptic clusters is discussed. The glycine receptor recently moved into the spotlight as a paradigm in the approach to cell biology of the formation of the postsynaptic membrane. PMID- 9394921 TI - Cell fate specification by localized cytoplasmic determinants and cell interactions in ascidian embryos. AB - Tadpole larvae of ascidians show the basic body plan of chordates. An ascidian larva consists of only a few types of cells and has a relatively small number of cells. Cell lineages are invariant among individuals and have been described in detail. These advantages facilitate the analysis of how the fate of each blastomere becomes specified during development. Over a century of research on ascidian embryogenesis has uncovered many interesting features concerning cellular mechanisms responsible for the fate specification. During embryogenesis, the developmental fate of a blastomere is specified by one of three different mechanisms: localized maternal cytoplasmic determinants, inductive interactions, or lateral inhibition in an equivalence cell group. PMID- 9394922 TI - Distribution of Na+,K(+)-ATPase in photoreceptor cells of insects. AB - Light stimulation of insect photoreceptors causes opening of cation channels and an inward current that is partially carried by Na+ ions. There is also an efflux of K+ ions upon photostimulation. Na+ and K+ gradients across the photoreceptor membrane are reestablished by the activity of the enzyme Na+,K(+)-ATPase. About two-thirds of the total amount of ATP consumed in response to a light stimulus is attributed to the activity of this ion pump, demonstrating the importance of this enzyme for photoreceptor function. Insect photoreceptor cells are polarized epithelial cells; their plasma membrane is organized into two domains having a distinct morphology, molecular composition, and function. The visual pigment rhodopsin and the molecular components of the transduction machinery are localized in the rhabdomere, an array of densely packed microvilli, whereas Na+,K(+)-ATPase resides in the nonrhabdomeric membrane. Comparative immunolocalization studies on compound eyes of diverse insect species have demonstrated subtle variations in the distribution patterns of Na+,K(+)-ATPase. These may be accounted for by differences in the mechanisms responsible for Na+,K(+)-ATPase positioning. PMID- 9394923 TI - Infertility and in vitro fertilization. A growing need for consumer-oriented regulation of the in vitro fertilization industry. PMID- 9394924 TI - Civil liability against prison officials for prescribing and dispensing medication and drugs to prison inmates. PMID- 9394925 TI - Medicine on trial. Physicians' attitudes about expert medical witnesses. PMID- 9394927 TI - Sudden (un)explained death. PMID- 9394926 TI - Tragic life or tragic death. Mandatory testing of newborns for HIV--mothers' rights versus children's health. PMID- 9394928 TI - Threshold stimulus ECT. PMID- 9394929 TI - Are insurance agencies making treatment decisions? PMID- 9394930 TI - Safety of combined pharmacotherapy. PMID- 9394931 TI - Safety of combined pharmacotherapy. PMID- 9394932 TI - Munchausen by proxy. PMID- 9394933 TI - Forensic child and adolescent psychiatry: a review of the past 10 years. AB - OBJECTIVE: To review important developments in child and adolescent forensic psychiatry from 1987 through 1996. METHOD: Major changes in the law and developments in research and practice were surveyed in the areas of the legal regulation of psychiatry, family law (divorce and child abuse), consultation to juvenile and criminal courts, civil litigation, and the development of the subspecialty. RESULTS: There has been a large increase in research based on quantifiable descriptive data of forensic populations, although studies using comparison or control groups remain relatively rare. While managed care has heavily influenced treatment practice, legal liability remains largely with the clinician. Issues regarding techniques of evaluation for sexual abuse have been scrutinized by the courts and by researchers. Legislative responses to rising rates of juvenile violence have been in the direction of treating violent adolescent offenders as criminally responsible adults. There has been a major move toward setting standards for forensic evaluations, training, and credentials. CONCLUSIONS: Child and adolescent forensic psychiatry remains an area encompassing diverse clinical issues. It remains unclear the extent to which it will develop into a formal subspecialty. PMID- 9394934 TI - Posttraumatic stress disorder in children: a review of the past 10 years. AB - OBJECTIVE: To review current knowledge about the clinical presentation, assessment, and treatment of posttraumatic stress disorder (PTSD) in children. METHOD: The literature on PTSD in children is examined. RESULTS: Over the past 10 years, PTSD has been described in children exposed to a variety of traumatic experiences. Little is known about the epidemiology of the disorder in children. Partial symptomatology and comorbidity are common. A variety of factors influence response to trauma and affect recovery. They include characteristics of the stressor and exposure to it; individual factors such as gender, age and developmental level, and psychiatric history; family characteristics; and cultural factors. Since the condition is likely to occur after disaster situations, much of the literature describes the child's response to disaster and interventions tend to include efforts within schools and/or communities. A number of clinical approaches have been used to treat the condition. CONCLUSIONS: While assessment has been studied extensively, the longitudinal course of PTSD and treatment effectiveness have not been. Biological correlates of the condition also warrant greater attention. PMID- 9394935 TI - Microanalysis of adolescent suicide attempters and ideators during the acute suicidal episode. AB - OBJECTIVE: To compare psychological and event-related contingencies that characterize and differentiate adolescent suicidal ideation and attempts. METHOD: Thirty-five ideators and 32 attempters (aged 12 to 17 years) consecutively referred to the suicide disorders clinic were evaluated with a semistructured interview about current and past emotional, cognitive, and behavioral states. RESULTS: Before the precipitant stressor (baseline), attempters reported significantly more hopelessness than ideators (odds ratio [OR] = 4.2, p < .05). During the suicidal episode, attempters, relative to ideators, spent more time ideating (OR = 4.3, p < .05), were more likely to isolate themselves (OR = 5.8, p < .01), and were less likely to tell anyone what they were thinking (OR = 4.5, p < .05). In contrast, ideators reported significantly more residual anger after the episode than did attempters (OR = 4.0, p < .05). All the episodes of ideation and attempts were preceded by a stress event. No differences were found between the groups on Beck Depression inventory scores. CONCLUSIONS: Preexisting hopelessness, a tendency toward isolation, not talking about ideation, and longer length of time ideating during suicidal episodes discriminated suicide attempters from suicide ideators. Knowledge of these factors may be helpful in preventive and treatment efforts with suicidal adolescents. PMID- 9394936 TI - Suicidal adolescents and ego defense mechanisms. AB - OBJECTIVES: To identify defense mechanisms that characterize adolescents with a range of suicidal behaviors and to differentiate them from nonsuicidal adolescents. METHODS: Fifty-five suicidal adolescent inpatients admitted for a definite suicide attempt were compared with 87 adolescent inpatients who had no history of suicide attempt or ideation and 81 nonpatients. Defense mechanisms were assessed by the Ego Defense Scale (EDS) which is part of a larger semistructured interview, the Child Suicide Potential Scale (CSPS), and by a self report questionnaire, the Life Style Index (LSI). The CSPS was also used to quantity violent and suicidal behaviors. RESULTS: On the LSI suicidal adolescent patients scored higher on denial, displacement, repression, and total defenses than the nonpatients. On the EDS they scored higher on regression, denial, projection, introjection, repression, and total defenses and lower on sublimation. LSI scores on displacement (higher) and on compensation (lower) distinguished suicidal from nonsuicidal inpatients. Denial and regression correlated positively and sublimation correlated negatively with both suicidal and violent behaviors. Introjection and repression correlated with suicidal behavior only. CONCLUSIONS: Overuse of displacement is connected with increased risk for suicidal and aggressive behaviors, while sublimation is probably a protective factor. In addition, several immature ego defenses possibly amplify aggression, which then is directed against the self by the maladaptive overuse of introjection, displacement, and repression. PMID- 9394937 TI - Suicidal ideation and behavior and noncompliance with the medical regimen among diabetic adolescents. AB - OBJECTIVE: To examine (1) the 1-year and lifetime prevalence of suicidal thoughts and behavior among adolescents with insulin-dependent diabetes mellitus (IDDM), (2) the relationship between suicidal thoughts and serious noncompliance with the medical regimen, and (3) factors including psychiatric disorder, self-efficacy expectations, and hopelessness that might mediate the relationship between suicidal thoughts and noncompliance. METHOD: Semistructured and structured interview instruments and self-report questionnaires were used to determine history of suicidal thoughts and behavior, serious noncompliance with the medical regimen, current psychiatric disorder, hopelessness, and self-efficacy expectations among 91 adolescents attending outpatient clinic appointments. RESULTS: The rate of suicidal ideation among the diabetic adolescents was higher than expected, but the rate of suicide attempts was comparable with that reported for the general population. Suicidal thoughts were strongly associated with serious noncompliance with the medical regimen. Duration of IDDM and psychiatric diagnosis were related to both suicidal ideation within the previous year and lifetime suicidal ideation. Diagnosable psychiatric disorder and not living in a two-parent home were related to noncompliance with medical treatment. CONCLUSIONS: Suicidal thoughts and serious noncompliance with the medical regimen are strongly associated among diabetic teenagers, and psychiatric disorder is a common correlate of both. PMID- 9394938 TI - Epidemiology of youth suicide in Israel. AB - OBJECTIVE: This study examined sociodemographic correlates and temporal trends in suicide among young Jews, Moslem Arabs, Druzes, and Christian Arabs in Israel. METHOD: The average yearly rates (1975 through 1989) for suicide and undertermined causes of death were calculated for children, adolescents, and subjects of army age. Rates were examined by gender, national/religious affiliation, and place of residence for all groups and, in addition, by ethnic origin among the jews. Logistic regression was used to ascertain the statistical differences. Temporal changes were examined by plotting the rates over the 15 year period and fitting a regression line. RESULTS: Among the young, differences by gender, national/religious, and ethnic origin did not consistently follow the pattern found in adults. Male Jews and Druzes of army age, facing a period of enhanced stress and availability of weapons, showed increased risk for suicide. Temporal trends for suicide differed from the worldwide pattern of increasing risk; these changes, however, were not homogeneous across all groups. CONCLUSIONS: The suicide rates among the youth in Israel, as in adults, are among the lowest in the world. Army service may be a period of enhanced risk, justifying preventive action. The almost worldwide trend of increasing suicide among the young is only partially present. PMID- 9394939 TI - Precipitating factors and life events in serious suicide attempts among youths aged 13 through 24 years. AB - OBJECTIVE: Precipitating factors and life events associated with medically serious suicide attempts were examined in young people making serious suicide attempts and control subjects. METHOD: Using a case-control design, the authors contrasted 129 young people making serious suicide attempts with 153 randomly selected community controls on a series of life event occurrences within the preceding year. Precipitating factors for serious suicide attempts were also examined. RESULTS: The most common precipitants of serious suicide attempts were relationship breakdowns, other interpersonal problems, and financial difficulties. However, one third of those attempting suicide were unable to describe any precipitating factor. Individuals who made serious suicide attempts had elevated rates of life events which were associated principally with interpersonal difficulties, work issues, financial difficulties, and legal problems. When due allowance was made for intercorrelations between life event measures and antecedent social, family, and personality factors, interpersonal losses and conflicts and legal problems remained significant risk factors for serious suicide attempts. CONCLUSIONS: Important proximal occurrences for serious suicide attempts among young people include a series of life events associated principally with interpersonal conflicts, relationship difficulties, and legal problems. PMID- 9394940 TI - Mania in children with pervasive developmental disorder revisited. AB - OBJECTIVE: Although a small literature of case reports suggests that mania co occurs with pervasive developmental disorder (PDD), little is known about this overlap. The authors systematically investigated the overlap between mania and PDD in a consecutive sample of referred youths, examining its prevalence and correlates. It was hypothesized that children with PDD plus manic features have both disorders. METHOD: Subjects were consecutively referred children meeting diagnostic criteria on structured interview for PDD without mania (n = 52), the comorbid condition PDD + mania (n = 14), and mania without PDD (n = 114). All subjects were evaluated using a comprehensive diagnostic battery that included assessment of psychopathology (structured diagnostic interview and Child Behavior Checklist), cognition, and functioning. RESULTS: Of the 727 referred children, 52 met criteria for PDD, 114 met criteria for mania, and 14 met criteria for both. The 14 children with both PDD + mania represented 21% of the PDD subjects and 11% of all manic subjects. Clinical characteristics of PDD were similar in PDD subjects with and without mania, and manic features were similar in manic children with and without PDD. CONCLUSIONS: Children with PDD and mania may suffer from two disorders. Comorbid mania among patients with PDD may be more common than previously thought. Identification of the comorbid condition may have important therapeutic and scientific implications. PMID- 9394941 TI - Autism and associated medical disorders in a French epidemiological survey. AB - OBJECTIVE: To estimate the prevalence of autism, to assess the strength of its association with specific medical disorders, and to test for a secular increase in its incidence. METHOD: An epidemiological survey was conducted among 325,347 French children born between 1976 and 1985 and living in three different French departements. Diagnosis, educational level, and associated medical conditions were abstracted from the records of children known to local educational authorities. Data were also pooled with those from another similar survey. RESULTS: One hundred seventy-four children (mean age: 11.6 years) with autism were identified. The prevalence rate was 5.35/10,000 (16.3/10,000 if other pervasive developmental disorders are included), with no difference according to geographical area or social class. Rates of medical conditions were as follows: 1.1% for tuberous sclerosis, 2.9% for chromosomal abnormalities including fragile X, 2.9% for cerebral palsy, 4.6% for sensory impairments, 0.6% for neurofibromatosis, 0.6% for congenital rubella, and 1.7% for Down syndrome. In the combined sample of 328 children with autism, the level and pattern of medical correlates were comparable, with tuberous sclerosis having a consistently strong association with autism. Prevalence rates were similar in successive birth cohorts. CONCLUSION: Medical disorders (excluding epilepsy and sensory impairments) accounted for fewer than 10% of the cases of autism. No secular increase in the prevalence of autism was found. PMID- 9394942 TI - Naltrexone in young autistic children: replication study and learning measures. AB - OBJECTIVE: This study expanded upon previous work on naltrexone efficacy and safety in young autistic children and assessed performance on learning measures. METHOD: Eleven children with autistic disorder, aged 3.0 to 8.3 years, were studied in home, school, and outpatient laboratory, bringing to 24 the combined study sample. Naltrexone, 1.0 mg/kg, was given daily in a randomized, double blind, crossover design. Dependent measures were parent and teacher Clinical Global Impressions (CGI) and Naltrexone Side Effects Rating Scale (SE), Conners Parent Impulsivity/Hyperactivity Factor, Teacher Hyperactivity Factor, laboratory CGI, and analysis of videotaped behavior. Learning measures were the Early Intervention Developmental Profile-Language and paired-associate learning. RESULTS: Comparisons between naltrexone and baseline, but not naltrexone and placebo, on parent and teacher ratings showed statistical significance. Three of 11 subjects improved in two or more settings. Side effects were mild. Administering naltrexone was a challenge. The combined study sample showed improvement on all parent measures and on Teacher CGI and SE-Restlessness compared with baseline and placebo. Eleven of the 24 children improved in two or more settings. Scores on learning measures did not change across conditions. CONCLUSIONS: Naltrexone was associated with modest improvement of behavior in 11 of 24 children, but learning did not improve. PMID- 9394944 TI - WISC profiles in child psychiatric diagnosis: sense or nonsense? AB - OBJECTIVE: WISC factor structure, the specificity of WISC factors, and diagnostic correlates of WISC profiles were studied to investigate the contribution of WISC profile analysis to child psychiatric diagnosis. METHOD: The fit of various factor models was tested and differences between various clinical groups regarding three WISC patterns were studied, using the WISC-RN (the Dutch version of the WISC-R) scores of a group of 465 Dutch children (mean age 11.2 years) referred to a psychiatric clinic. RESULTS: The traditional factor models were replicated in this study. However, most of the variance in the factors could be explained by an underlying factor, "general intelligence," suggesting that WISC factors measure specific cognitive abilities only to a limited degree. Another important finding is that the various DSM-III-R and Child Behavior Checklist diagnoses could not be distinguished on the basis of WISC profiles. CONCLUSION: The data demonstrate that the relationship between WISC factors and specific cognitive abilities and neuropsychological functions needs further clarification in order to improve the validity of the traditional use of WISC profiles as a source of diagnostic information. PMID- 9394943 TI - Neurological soft signs: one-year stability and relationship to psychiatric symptoms in boys. AB - OBJECTIVE: This study had two main objectives: (1) to examine the 1-year stability of neurological soft signs and (2) to examine the longitudinal relationship between soft signs and psychiatric symptoms in young boys. METHOD: A consecutive series of 56 boys from a high-risk sample received standardized psychiatric and soft sign assessments at study intake. Approximately 1 year later, 48 (86%) of these boys received a reassessment of their psychiatric and soft sign status. RESULTS: Soft signs exhibited marked stability across the 1 year period (intraclass correlation = .70, p < .001). Symptoms of both internalizing and externalizing disorders correlated with poor performance on the soft sign examination. For both internalizing and externalizing symptoms, the association with soft signs occurred primarily among individuals with persistently high scores on symptom scales across the two assessments. CONCLUSIONS: Performance on a standardized neurological soft sign examination is stable over a 1-year period. Soft signs measured with this examination relate to both internalizing and externalizing symptoms in young boys, particularly when symptoms are relatively stable over time. Further research should consider the clinical significance of childhood soft signs. PMID- 9394945 TI - Traumatic brain injury in a child psychiatry inpatient population: a controlled study. AB - OBJECTIVE: To extend our findings from child psychiatry outpatients to child psychiatry inpatients regarding the similarity of children with a history of traumatic brain injury (TBI), particularly mild TBI, to matched children without such a history. METHOD: This is a chart review of patients consecutively admitted to a child psychiatry inpatient unit over a 5-year period. Children with TBI were matched by age, sex, race, and social class to children with no history of TBI. Axis I and II diagnoses and diagnostic clusters and use of special education services and IQ scores were compared. RESULTS: Fifty-six (8.1%) of 694 consecutive patients admitted had a definite TBI. Not one of more than 50 variables compared between TBI and control subjects was significantly different. CONCLUSION: In a child psychiatry inpatient unit, patients with a history of TBI were virtually indistinguishable from matched children without TBI. Caution should be exercised before attributing the child's problems, especially long-term problems, to the TBI unless the injury was severe or the child is exhibiting related phobic or posttraumatic stress symptomatology. PMID- 9394946 TI - Case study: trauma-related hallucinations. AB - Proper differential diagnosis of psychiatric disorders with psychotic symptoms is imperative, as the treatment implications of the various conditions are quite different. A case study of a 5-year-old abused child with posttraumatic stress disorder is presented to illustrate some of the characteristic features of psychotic symptoms in traumatized children. Literature reviewed suggests that trauma-related hallucinations frequently contain content which is related to children's life experiences, are exacerbated by "triggers" and safety concerns, resolve with psychotherapy or psychosocial interventions, and are resistant to standard neuroleptic treatments. They are also associated with unique clinical, familial, developmental, and psychobiological correlates, and they require multifaceted treatment interventions. PMID- 9394947 TI - Case study: neuropsychiatric symptoms associated with the antimalarial agent mefloquine. AB - The development of acute neuropsychiatric symptoms in a 10-year-old boy subsequent to his return from travel abroad in Africa, where he had taken the antimalarial agent mefloquine (Lariam), is reported. A 4-week course of cognitive behavioral therapy was used to effectively treat this substance-induced anxiety disorder, which had been caused by treatment with mefloquine. A review of the literature about adverse neuropsychiatric effects of mefloquine and the differential diagnosis of malaria is provided. In an age in which international travel is occurring with increasing frequency, it is important to obtain travel histories, including exposure to prophylactic medication, when patients present with acute-onset psychiatric symptoms. PMID- 9394948 TI - Adolescents with psychiatric disorders and the risk of HIV. AB - OBJECTIVE: To review literature relevant to human immunodeficiency virus (HIV) associated risk behaviors among adolescents with psychiatric disorders and psychological influences on risk behaviors. METHOD: This report is based on review of 66 articles, which comprise all of the relevant literature in the English language. RESULTS: Although the seroprevalence of HIV in adolescents with psychiatric disorders is unknown, studies indicate that adolescents with psychiatric disorders are at greater risk than their peers because of increased rates of unsafe sexual practices, impulsivity, self-destructive attitudes, cognitive immaturity, high rates of substance use, self-cutting behavior, and the sequelae of sexual abuse. CONCLUSION: Directions are proposed for the design of developmentally appropriate, clinically oriented HIV prevention interventions based on the relationships between psychological dysfunction, social stressors, and HIV risk behaviors. PMID- 9394949 TI - Tourette's syndrome and psychopathology in a child psychiatry setting. AB - Prior investigations of psychopathology among children with Tourette's syndrome (TS) have rarely used child psychiatry samples and sophisticated personality instruments. OBJECTIVE: To produce an objectively derived composite TS personality profile and to determine the rate of particular problems in a TS psychiatry sample compared with children with out TS from the same clinical practice. METHOD: Children (n = 33) referred to child psychiatrists because of emotional and behavior problems who were subsequently also found to meet DSM-III R criteria for TS were assessed by the Personality Inventory for Children. RESULTS: Children with TS expressed high rates of psychopathology overall (composite 2.7 SD elevated) with depression, anxiety, and peculiar behavior having the highest values; depression occurred most frequently (73%), and attention-deficit hyperactivity disorder (55%) was no more common than among comparison group children and conduct problems (18%) were rarer. "Depression, anxiety, tension, and excessive worry" were characteristic of the actuarially derived modal TS personality. CONCLUSIONS: The prevalence and manifestations of psychopathology of children with TS in a child psychiatry practice are not identical with those reported in the literature. Child psychiatrists should be particularly vigilant of depressive symptoms and expect to encounter relatively few conduct problems compared with children without TS. Establishing "local prevalence rates" for children with TS seeking psychiatric evaluation can help guide the diagnostician and make diagnosis more assured. PMID- 9394950 TI - Hopelessness in inpatient youths: a closer look at behavior, emotional expression, and social support. AB - OBJECTIVE: To examine the individual and family characteristics of children and adolescents with high levels of hopelessness. METHOD: One hundred inpatient youngsters participated in the study. Several measures, including the Hopelessness Scale for Children, Problem Behavior Scale of the Scales of Independent Behavior, Social Support Questionnaire-Revised, Pediatric Anger Expression Scale, and Differential Emotions Scale, were used to compare differences between youngsters who scored high or low on hopelessness. RESULTS: The results indicated that youngsters with high hopelessness scores tended to perceive their families and peers as providing little support, to express their anger overtly and aggressively, and to demonstrate more negative emotions than youngsters with low hopelessness scores. CONCLUSIONS: Hopelessness in youths appears to be associated with a specific pattern of behavioral and emotional problems. Clinical implications of the findings include integrating anger management, emotional expression interventions, and involving the family in treatment to enhance the social support network of youngsters with high levels of hopelessness. PMID- 9394951 TI - Treating anorexia nervosa patients in the era of managed care. PMID- 9394952 TI - Altered kidney matrix gene expression in early stages of experimental diabetes. AB - The expression of mRNA and distribution of alpha 1(IV), alpha 3(IV) chains of type IV collagen, matrix metalloproteinase 2 (MMP-2), and tissue inhibitor of metalloproteinase 1 (TIMP-1) were examined in kidneys from streptozotocin diabetic rats, 2.5 months after administration of the drug, an early time point when specific diabetic glomerular changes were still minimal. Ten age-matched Sprague-Dawley rats were assigned to control and diabetic groups. Compared to the controls, the diabetic rats had a significantly lower body weight, higher kidney weight and serum glucose levels, but no significant changes of glomerular surface area and urine albumin were observed. Northern blot analysis, using whole kidney mRNA, revealed that diabetic rat kidneys expressed 113.5% more alpha 1(IV), 46.5% more alpha 3(IV), 54.8% less MMP-2 and 246% more TIMP-1 (in all instances: p < 0.05). These results were corroborated by in situ hybridization for RNA expression. A quantitative analysis of the data indicated the following changes in glomeruli: (1) 74.6% more alpha 1(IV), (2) 103.8% more alpha 3(IV), (3) 40.7% less MMP-2 and (4) 80.9% more TIMP-1. Similar changes were observed in tubular (proximal and distal) cells. We conclude that an increased synthesis and decreased degradation of renal extracellular matrix components occur early after induction of experimental diabetes, before the onset of typical structural changes in the kidneys, and represent changes of specific gene expression at the transcriptional level. All the cell types in the glomerulus as well as the proximal and distal tubules appear to be involved in this alteration of expression, and this is a novel finding. PMID- 9394953 TI - Dynamics of endothelium-muscle cell contacts in the coronary artery of the dog in ontogeny. AB - The myo-endothelial area in the coronary artery conduit was described in 3 developmental stages: in fetuses, newborns, and adult dogs. Transmission and scanning electron microscopy were used for the study, and morphometry was used for quantitative evaluation. In all three stages, the internal elastic lamina was found to be fenestrated. Endothelial cells and smooth muscle cells (SMC) approached the fenestrae, and protrusions of one or both cells entered into the fenestrae. In some places contacts between endothelial and SMC were found. The patterns of mutual approaches of smooth muscle and endothelial cells, as well as the entering into the fenestrae were similar in all three stages. The myo endothelial contacts were counted per 100 microns inner circumference of the coronary artery and the numbers observed, i.e. 5.17 +/- 0.50 in fetuses, 1.94 +/- 0.17 in newborns and 0.33 +/- 0.09 in adult animals, proved clearly that the frequency of myo-endothelial contacts, highest in fetuses, decreases with age. With regard to the dual control of the coronary smooth muscle and/or diameter, it is noteworthy that an opposite trend can be observed in the development of innervation of the coronary artery: the autonomic nerve fibres with varicosities are missing in the coronary wall 1 week before birth, while after birth their number keeps increasing. Remarkable enough is also the difference in distances between the endothelium and SMC on the one hand and nerve varicosities and SMC on the other. The above facts indicate a prevalence of endothelial control of coronary diameter. PMID- 9394954 TI - Organization of the lamina propria mucosae of rat intestinal mucosa, with special reference to the subepithelial connective tissue. AB - Light microscopy, scanning electron microscopy and transmission electron microscopy have been used to delineate the structure and function of the lamina propria mucosae in the rat jejunum. In silver-impregnated sections, the adepithelial surface of the lamina propria mucosae was framed by a sheet of reticular fibers (reticular sheet). Short-term (3-hour) immersion of jejunal tissues in 2 N NaOH solution enabled us to simultaneously view networks of reticular fibrils and fibroblasts residing in the subepithelial connective tissue under a scanning electron microscope. The reticular fibrils, which measured about 40 nm in diameter and were interwoven in dense networks, formed a sheet 2-3 microns thick. In the villi, this sheet contained numerous foramina ranging from 3 to 7 microns in diameter, through which lymphocytes, macrophages, basal extensions of epithelial cells and fat particles traversed. The reticular sheet in the domes of isolated lymphoid nodules was markedly porous, and many lymphocytes migrated into or out of the epithelium through the foramina. The formaina of the reticular sheet may participate in the communication between the intestinal epithelium and the lamina propria mucosae. It was noted that the foramina of the reticular sheet in the villi were surrounded by end feet of the cytoplasmic processes of fibroblasts. In addition, these fibroblasts were combined with lymphocytes or dendritic cells in the lamina propria mucosae. PMID- 9394955 TI - Long-term effects of core decompression by drilling. Demonstration of bone healing and vessel ingrowth in an animal study. AB - Avascular necrosis of the femoral head is associated with bone marrow hyperpression. Although core decompression by drilling is an accepted treatment regimen, until today no experimental results exist concerning the physiological effects of this procedure. Published clinical data are controversial. In an animal study marrow decompression was carried out by drilling of both hips in 18 healthy male sheep. In the right hip of each animal a resorbable stent was implanted in order to prolong the duration of core decompression. Over a time period of 24 weeks the effects were studied by measurement of the intraosseous pressure, by the plastination method and by morphological examination with light and electron microscopy. Bone drilling is a procedure of high short-time efficacy in decompressing the bone marrow. But decompression lasts only for a short time period. Three weeks postoperatively the drill channel is sealed by hematoma and fibrous tissue in both hips (with/without stent) and no significant decompressive effect is measured. Ingrowth of vessels along the drill channel is found in all hips after a time period of 3 weeks. These vessels originate from the periosteum as well as from the bone marrow and form temporary anastomoses between the periostal-diaphyseal-metaphyseal and the epiphyseal-physeal circulatory system. In conclusion, for the first time an anastomosis induced by drilling between both circulatory systems of bone is demonstrated and the importance of the periosteum is confirmed. The time of decreased core pressure induced by drilling is too short for substitution of a necrotic area and could be the explanation of the inferior clinical results of the procedure. PMID- 9394956 TI - Quantitative analysis of incongruity, contact areas and cartilage thickness in the human hip joint. AB - Joint incongruity and cartilage thickness have been shown to determine the contact stresses and the load partitioning between the solid and fluid phases of articular cartilage. Matrix stresses, which are relevant in the development of osteoarthrosis, can, however, not be determined experimentally but must be calculated using numerical methods. The aim of the present study was to quantify the incongruity and cartilage thickness of the human hip, in order to allow for the construction of morphologically accurate finite element models. Twelve cadaveric specimens (34-86 years), two fresh and ten fixed, were investigated. The loading configuration was based on in vivo measurements of hip joint forces during midstance. The incongruity and contact areas were determined using a polyether casting technique, in the minimally and the fully loaded state. The cartilage thickness was measured at identical coordinate points with an A-mode ultrasonic system. Generally, the contact started at lower loads at the edge of the lunate surface, and the joint space increased towards its central aspects. In some specimens the contact started in the acetabular roof, leaving a joint space of up to 2 mm in the horns of the lunate surface. In others, the initial contact was observed in the anterior and posterior horns of the lunate surface with a joint space width of up to 0.75 mm in the acetabular roof. The size of the contact areas increased from about 20% of the lunate surface to 98% at higher loads. The articular cartilage thickness ranged from 0.7 to 3.6 mm, the maxima being located in the ventral aspects of the femoral head and acetabulum. These quantitative data on joint space width, contact, and cartilage thickness in the human hip joint may be used to construct and validate finite element models which are required to elucidate the mechanical factors involved in osteoarthrosis. PMID- 9394957 TI - Blebs in the mouse cerebellar granular layer as a sign of structural inhomogeneity. 1. Anterior lobe vermis. AB - The cerebellum is a modular structure. However, the size of the fundamental compartments is uncertain, with anatomical methods showing a parasagittal band arrangement but electrophysiological mapping suggesting a finer subdivision into microzones and patches. A new anatomical way to demonstrate compartmentation is described. The cerebellum is fixed by perfusion with 70% ethanol, paraffin embedded and sectioned. When the sections are rehydrated the granular layer pleats into an elaborate array of blebs. These blebs are seen in both transverse and sagittal sections, found in all lobules of both the vermis and the hemispheres, symmetrical about the midline, reproducible between neighboring sections and between individuals, and bear a constant relationship to the Purkinje cell bands as revealed by zebrin II immunocytochemistry. The data suggest that the granular layer of the adult mouse cerebellum is divided into several thousand modules. These modules may reflect the mossy fiber topography, and may be the anatomical equivalents of the tactile receptive field patches. Such a profound compartmentation has important implication for theories of cerebellar structure and development. PMID- 9394958 TI - Magnetic resonance imaging of cerebral associative white matter bundles employing fast-scan techniques. AB - Rapid scan techniques have introduced new sequence parameters as well as novel contrast concepts into everyday magnetic resonance imaging (MRI). In particular contrast characteristics of fast-spin echo (FSE) sequences showed some significant differences when compared to conventional spin echo images. The purpose of this work was to demonstrate the capabilities of FSE MRI in identifying and characterizing the in vivo anatomy of the main cerebral associative systems. Between March and November 1995, 20 healthy adult volunteers (12 males, 8 females, mean age 35 years) were submitted to a cranial MRI examination (1.5 Philips Gyroscan NT). In all cases axial and coronal 2 dimensional FSE T2-weighted and 2-dimensional inversion recovery FSE T1-weighted images were obtained. All MRI images were examined by a neuroradiologist (G. Dal Pozzo) for the depiction of the following compact white matter fiber bundles: anterior commissure, corpus callosum, superior fronto-occipital fasciculus, cingulum, fornix, mammillothalamic tract, uncinate fasciculus, superior and inferior longitudinal fasciculus. All these associative pathways could be well identified on T2-weighted images due to a lower signal intensity with respect to the surrounding white matter. On T1-weighted images only the corpus callosum, the anterior commissure and the fornix could always be identified. Correlation with myelin-specific colorations (Luxol fast blue stains) in anatomic atlases and a review of the literature on the myelinization process during infancy indicate that the short T2 relaxation times of the aforementioned cerebral associative systems may be due to heavy myelination and high fiber density. The correct visualization of interintrahemispheric associative white matter fiber bundles may play an important role in white matter disorders like dys- and demyelinating diseases and in the spreading of vasogenic edema and/or tumor being useful for their staging. PMID- 9394959 TI - Anterior tympanic artery: course, ramification and relationship with the temporomandibular joint. AB - The anterior tympanic artery, a branch of the maxillary artery, ascends through the retroarticular region dividing into anterior branches that spread through the posterior part of the temporomandibular joint, and posterior branches that contribute to the vascularization of the external acoustic meatus and the tympanic cavity. The arrangement of the anterior tympanic artery was studied bilaterally in 18 adult cadavers. In some cases, the anterior tympanic artery branches off from the superficial temporal artery. The relationships of the anterior tympanic artery with the posterior part of the temporomandibular joint were analyzed. PMID- 9394960 TI - The management of malignant melanoma revisited. PMID- 9394961 TI - Bacterial contamination of pneumoperitoneum gas in peritonitis and controls: a prospective laparoscopic study. AB - There is a theoretical risk that the pneumoperitoneum gas could carry bacteria in aerosol form and spread infection throughout the peritoneal cavity during laparoscopy for infective conditions such as appendicitis. The aim of this study was to attempt to culture bacteria from the pneumoperitoneum gas during laparoscopy for potentially infected cases and a group of controls. A total of 53 consecutive laparoscopies were studied, of which 21 were potentially infected and 32 served as controls. A lavage of the operative site was positive for pathogenic bacteria in almost 30% of the potentially infected group and only 3% of the control group. The pneumoperitoneum gas was bubbled through blood culture medium at the beginning and the end of the procedure, but only one of the 106 bottles grew any bacteria, and the specimen was a likely contaminant. In conclusion, we were unable to grow any significant bacteria from any of our cases despite using a sensitive method and demonstrating pathogenic bacteria in the peritoneal lavages. The pneumoperitoneum itself is unlikely to disperse bacteria. PMID- 9394962 TI - Caecostomy in the management of acute left colonic obstruction. AB - In the management of acute left colonic obstruction there is a tendency to perform immediate resection with anastomosis. We evaluated 27 consecutive patients (mean age 73.8 years) with acute left colonic obstruction and gross dilatation of the proximal colon treated by the "traditional" staged procedure. After caecostomy, no further resection was performed in two patients. In 25 patients, the obstructing tumour was resected after a median period of 14 days. In 17 (68%) patients the caecostomy was closed simultaneously. In 8 patients this was done at a third stage. Histologic examination revealed diverticular disease in 6 and adenocarcinoma in 19 patients. No deaths occurred after caecostomy nor was there major morbidity. After colonic resection, one in-hospital, nonprocedure related, death occurred (mortality rate 4%). In 21 patients with an anastomosis no dehiscence occurred. Other postoperative complications occurred in 5 patients (morbidity rate 20%). The median hospital stay for patients with a two-stage procedure was 32 days and with a three-stage procedure 39.5 days. The staged procedure in the management of acute colonic obstruction is still a safe and acceptable procedure in elderly patients with acute large bowel obstruction. To shorten the hospital stay the period between caecostomy and colonic resection should be reduced and it is best to close the caecostomy simultaneously. PMID- 9394963 TI - Carotid chemodectomas. Experience with nine cases with reference to preoperative embolization and malignancy. AB - The medical records of nine patients (five female and four male, mean age 58 +/- 5 years) presenting with a carotid chemodectoma between 1983 and 1995 were reviewed. In two cases (22%) the diagnostic was not suspected at the time of initial presentation. The most common complaint was a swelling in the anterolateral region of the neck. One patient (11%) presented with a preoperative peripheral nerves deficits (vagus and hypoglossal palsies and Horner's syndrome). Two tumours were embolized preoperatively with polyvinyl alcohol particles. Complete surgical excision was possible in each patient and the plane of resection was adventitial. In three cases, early ligation of the external carotid artery facilitated the resection. In two patients, the vagus nerve was sacrificed because of tumour involvement. No operative mortality was observed and no vascular complication occurred. In addition to the patient with preoperative neurologic symptoms, three patients developed peripheral nerve deficits (vagus and hypoglossal nerves) postoperatively. Two of these deficits were transient. These peripheral neurologic complications were observed with the largest tumour sizes. Two cases were malignant (lymph nodes and bony metastases). These two patients received postoperative radiotherapy. The mean follow-up period 63 +/- 19 months. No patient developed local recurrence during the follow-up. Two patients died during the follow-up, one for condition unrelated to their disease and the second from metastatic dissemination. In conclusion, carotid chemodectomas may be safely resected. The best way to minimize the rate of complications is to operate them at an early stage of evolution. PMID- 9394964 TI - Thrombangiitis obliterans (Buerger's disease): still a limb threatening disease. AB - A series of 29 well-documented and properly analysed patients with thrombangiitis obliterans (Buerger's disease) is presented. The diagnosis of Buerger's disease was based on following criteria: smoking history, onset before the age of 50 years, infrapopliteal arterial occlusive disease, either upper limb involvement or phlebitis migrans, absence of atherosclerotic risk factors other than smoking. In the last 10 years (1986-1996), we identified 29 patients who met these rigid criteria. There were 24 men and 5 women, aged 32.4 years at the moment of the disease first clinical symptom. The cumulative tobacco use averaged 16 pack-years for each patient. The initial symptom was limited gangrene of a toe (n = 9) or a finger (n = 2), foot claudication (n = 6), calf claudication (n = 3), rest pain (n = 3), migratory superficial phlebitis (n = 4), and Raynaud phenomenon (n = 2). Angiography and/or Doppler ultrasound revealed digital, pedal and calf artery involvement in all patients, with proximal extension in ten patients (femoropopliteal in ten, including three cases with external iliac artery involvement). Seven patients had also evidence of upper limb involvement. Histologic proof was available in only seven patients. Only nine patients completely stopped smoking. Treatment was exclusively medical in five cases. Twenty-four underwent sympathectomy (20 at lumbar, and four at thoracic level), with good immediate result in 16. In 11 patients a vascular reconstruction was done (eight femorocrural and three iliofemoral bypasses), with a patency rate of only 36% at two years. Amputation was required in 16 patients (a mean of 2.7 amputations per patient) at one or more levels: toe (n = 19), forefoot (n = 5), below knee (n = 8), above knee (n = 2), finger (n = 3). Two patients ended up with bilateral leg amputation. Overall, 23% (7/30) of the patients required major leg amputation during the course of the disease. Disease progression was moderately related to continued tobacco use. Buerger's disease still entails considerable risk of major amputation. Complete abstinence from tobacco use is crucial to expect stabilization of the process. However, in advanced stages of the disease and despite cessation of smoking recurrent episodes of ischaemia or tissue loss are not excluded. PMID- 9394965 TI - Surgical approaches to the humeral shaft. AB - Open fractures, transverse, short oblique and spiroid fractures of the humeral shaft, as well as comminuted fractures with radial palsy or vascular injury, mostly lead to bad end-results if treated conservatively. The same is valid in the case of bilateral humeral shaft fractures, multiple injuries, polytrauma, pathologic fractures and pseudarthrosis. Good end-results and a low rate of complications in the operative procedure require an adequate approach to the fractured limb as well as a meticulous care of the soft tissues. In plate osteosynthesis, the anterolateral approach for the proximal third of the shaft, the anterolateral approach with radial exposure for the middle third of the shaft and the posterior approach for the distal third of the shaft seem to offer the best pathway for reposition and fixation, respecting the biologic requirements for a successful osteosynthesis. The approaches for external fixation demand a thorough knowledge of the course of the axillary and radial nerves. Unreamed intramedullary nailing can be done in an anterograde and in a retrograde way. In anterograde nailing, damage of the rotator cuff must be avoided, in retrograde nailing, the elbow capsule should be left closed and untouched. PMID- 9394966 TI - The diagnosis by fine needle aspiration biopsy of hydatid cyst of the pancreas. AB - We describe a case of pancreatic hydatid cyst in which the definitive diagnosis was made at fine needle aspiration cytology. The most common site of hydatid cyst is the liver (65%), followed by the lungs (25%). The hydatid cyst of the pancreas is rare since it accounts for less than 1% of the various sites of hydatid disease. The diagnosis may be difficult when the presentation is that of an unexplained epigastric mass or cyst, despite suggestive radiological and ultrasonographic features. Modern serology tests are positive in up to 80% of the abdominal hydatid cysts. It is mandatory to obtain a fine needle aspiration biopsy for definite diagnosis and for appropriate treatment planning. Surgical removal of the hydatid cyst still remains the most effective treatment. PMID- 9394967 TI - Mesenteric venous thrombosis. Diagnostic and therapeutic approach. AB - Mesenteric venous thrombosis is an infrequent but distinct form of intestinal ischaemia. We report a case of acute mesenteric venous thrombosis diagnosed by computed tomography. Laparoscopy permitted to establish the extent of the ischaemia. Initially, high doses streptokinase were administered for 6 hours, followed by heparinotherapy for 10 hours, with the aim to reduce the length of bowel to be resected. One day later, intestinal resection was carried out, followed by postoperative anticoagulation. PMID- 9394968 TI - Spinal ischaemia after surgery for abdominal infrarenal aortic aneurysm. Diagnosis with nuclear magnetic resonance. AB - A 76-year-old man underwent surgery for an infrarenal aortic aneurysm reaching 6 cm in maximal transverse diameter. The aorta was crossclamped below the level of the renal arteries. A tube graft was interposed and tend between the infrarenal aorta and the aortic bifurcation. Due to leakage on the suture line two consecutive episodes of crossclamping for a total duration of 40 min. were required. No hypotension was noted during or after the procedure. After operation, the patient complained of difficulties to move both legs and neurologic examination demonstrated paraparesis, with mild sensory deficit. Faecal and urinary incontinences were also noted and urodynamic testing demonstrated sphincterovesical palsy. Nuclear magnetic resonance imaging detected an ischaemic zone in the spinal cord at the level of T11. Faecal incontinence and motor deficit partially resolved but no bladder function recovery was observed. Spinal ischaemia is a rare complication after abdominal aortic surgery. Several risk factors have been suggested which include level and duration of the aortic crossclamping, possible interruption of the spinal cord blood supply via the greater medullary artery (the so-called artery of Adamkiewicz), presence of intra or postoperative episodes of hypotension, atheromatous embolization, underlying occlusive arteriosclerosis of spinal arteries, and respect or not of the hypogastric circulation. In our case, the duration of the crossclamping and interruption of the blood flow in lumbar arteries probably supplying the distal spinal cord were likely contributive factors. PMID- 9394969 TI - Ulnar artery false aneurysm: temporary ultrasound-guided compression closure in an unusual case. AB - A case of traumatic false aneurysm located in an unusual portion of the ulnar artery is presented and the clinical findings, diagnosis and treatments are considered. The authors report the first use of ultrasound-guided compression closure, as an alternative to surgical management of ulnar artery false aneurysms. PMID- 9394970 TI - Osteosarcoma of the proximal fibula: report of 3 cases. AB - Osteosarcoma of the proximal fibula represents about 2% of all osteosarcomas. The complexity of anatomical structures in this region led in the past to low transfemoral amputation in order to achieve local control. Three patients aged 14, 13 and 14 were treated in our institution and surgical margins were free of tumour after a wide en bloc resection. To achieve that, the lateral wall of the tibia and the anterior tibial artery were taken out with the proximal fibula. All three patients are alive without evidence of disease. The functional results are evaluated according to Enneking system recommended and adopted by the Musculo Skeletal-Tumor Society (M.S.T.S). Overall functional results were 80, 100 and 100%. PMID- 9394971 TI - Non-invasive in vivo techniques to differentiate photodamage and ageing in human skin. AB - It is important to differentiate skin changes due to the intrinsic ageing process from those due to chronic photodamage in the development of therapies to assist the latter condition, and in this study we have used instrumental techniques to differentiate between changes in a range of properties of skin due to ageing and those due to photodamage, especially with regard to elasticity. We measured three sites of differing sun exposure in a group of younger and in a group of older subjects. A pulsed A-scan ultrasound system was used to measure skin thickness, and a uniaxial extensometer was employed to assess elastic properties. Skin surface roughness measurements were made using silicone rubber impressions and a stylus profilometer. We demonstrated significant differences in skin roughness between young and old subjects at every site and differences between sun-exposed and sun-protected sites only in the older group. Parameters of the elastic properties of skin differed between the groups, and also between sites of most different sun exposure. The uniaxial extensometer can demonstrate a loss of the skin's elasticity predominantly by photodamage, and the roughness of the skin surface can be shown to increase mostly by chronological ageing but to decrease modestly by photodamage. This demonstrates that differences between the two processes can be quantified, and indeed they should be. PMID- 9394972 TI - Quantification and specificity of the repeated open application test (ROAT). A methodological study using cobalt and colophony in guinea pigs. AB - The repeated open application test is used to assess the clinical relevance of positive patch test reactions to ingredients of formulated products. The great variation in outcome is usually claimed to be related to the concentration of the allergen responsible. We have here studied the quantitative aspects, specificity and effect of patch testing on the outcome of the repeated open application test in an animal model, using guinea pigs sensitized with cobalt chloride or colophony. Thresholds of sensitivity were determined before and after the topical treatments. Clear dose-response relationships were established. The reactivity in sham-treated controls and to the vehicles was minimal. The concordance between patch test results and outcome of the use tests was concentration-dependent and at low concentrations < 50%. The repeated open application test is a useful method, but some of the basic issues need further evaluation. This animal model will hopefully serve this purpose. PMID- 9394973 TI - Flow cytometric DNA content analysis of ultraviolet light-induced squamous cell carcinomas: a comparative study of squamous cell carcinomas of the lip and those arising from other sites of sun-damaged skin. AB - The development of squamous cell carcinoma (SCC) of the lip is considered to be mainly related to excessive sun exposure. However, its higher metastatic rate is distinct from that of SCCs arising from other sites of sun-damaged skin. Flow cytometric DNA content analysis, using paraffin-embedded specimens, was performed on 15 SCCs of the lip and 32 SCCs arising from other sites of sun-damaged skin. A significantly lower incidence of DNA-aneuploidy was observed in SCCs of the lip (2/15) than in those of sun-damaged skin (15/32) (p < 0.05). The mean age of patients with SCC of the lip (66.7 +/- 11.6 years; mean +/- SD) was significantly lower than that of the other patients (78.1 +/- 11.1 years) (p < 0.01). There was no significant difference between the mean diameter size of tumors on the lip (19.5 +/- 5.7 mm) and that of tumors on other sites of sun-damaged skin (30.7 +/- 20.5 mm). These results suggest that additional carcinogenic factors besides ultraviolet light may be involved in the development of SCC of the lip. PMID- 9394974 TI - Keratinocytes cultured under hyperthermal conditions secrete factor(s) which can modulate dermal fibroblast proliferation and extracellular matrix production. AB - We have studied how keratinocytes cultured under hyperthermal conditions modulate skin fibroblast growth potential and their biosynthetic phenotypes in vitro. When keratinocytes were cultured at 30, 34, 37 or 39 degrees C, the conditioned medium of the keratinocytes cultured at 39 degrees C showed a greater inhibitory activity for fibroblast proliferation and greater synthetic activities of collagen and glycosaminoglycans than those incubated at 30, 34, or 37 degrees C. Transforming growth factor (TGF) beta 1 production in skin fibroblasts was also stimulated by the keratinocyte conditioned medium cultured at 39 degrees C. The stimulating activity of collagen and glycosaminoglycan syntheses of keratinocyte conditioned medium may be explained at least partly by enhanced TGF beta 1 production. The results indicate that keratinocytes cultured at a higher temperature (39 degrees C) may secrete factor(s) which modulate both fibroblast growth and matrix synthesis. This may provide evidence that under hyperthermal conditions epidermis can influence the functions of skin fibroblasts. PMID- 9394975 TI - Evaluation of scratch movements by a new scratch-monitor to analyze nocturnal itching in atopic dermatitis. AB - Itching is very important in atopic dermatitis, but the details of itching or scratch movements, especially during sleep at night, have not yet been fully comprehended. We designed a new, simple device, the Scratch-Monitor (SM), to evaluate scratch movements at night and assessed the usefulness of this device by a comparison involving 26 patients and 17 healthy controls. The SM, a box weighing only 25 g with a pressure sensor on the bottom, is attached to the back of each hand under a cotton glove and records the number as scratch movements per minute in the case of more than three successive changes of pressure. The SM indicated that patients with atopic dermatitis scratched more frequently and suffered more severe sleep disturbance than healthy controls. Although the SM had several problems related to specificity and sensitivity, we conclude that the SM is a useful tool for evaluating nocturnal itching. PMID- 9394976 TI - Inducible nitric oxide synthase demonstrated in allergic and irritant contact dermatitis. AB - Eight allergic patch test reactions, eight irritant skin reactions induced by 3% sodium lauryl sulphate and six normal controls were biopsied. Biopsies were immunohistochemically stained with a mouse monoclonal antibody to inducible nitric oxide synthase (iNOS), and staining was quantified by computerised image analysis. Human chondrocytes induced to express iNOS were used as a positive control. A significant increase in iNOS was found in both irritant and allergic contact dermatitis. There were no differences in the distribution of expression of iNOS. The antibody used was confirmed by Western blotting not to cross-react with the endothelial isoform of nitric oxide synthase (NOS) but did cross-react with a 150 kDa protein, which may be neuronal NOS or an isoform of neuronal NOS. PMID- 9394977 TI - Upregulation of CD40 and CD40 ligand expression in IgE-associated cutaneous diseases. AB - In order to better understand the immunological processes connected with IgE associated cutaneous disease, we have examined the expression of CD40 and its ligand CD40L, required for the induction of IgE synthesis in B-cells, as well as of IgE and its receptors in various dermatoses (atopic dermatitis (AD), scabies, chronic recurrent urticaria) versus normal skin, and in one dermopathic lymph node versus normal lymphatic tissue by immunohistochemistry. Compared to normal skin, cells expressing IgE, Fc epsilon RI, Fc epsilon RII, CD40, CD40L and L26 were increased in the dermis, partly also in the epidermis, from patients with AD and scabies, but not in chronic urticaria. CD40 and CD40L were detected on numerous cells in lymphatic tissue from both normal donors and a patient with AD, whereas large numbers of IgE- and Fc epsilon RI-positive cells were only found in the dermopathic lymph node from the AD patient, in contrast to very few in normal lymphatic tissue. These results with selectively increased IgE/Fc epsilon RI and associated CD40/CD40L expression in the skin of AD and scabies suggest that cutaneous tissue, in addition to dermopathic lymphatic tissue, might contribute to IgE synthesis. PMID- 9394978 TI - Sleep patterns in children with atopic eczema. AB - Following routine appointments in a paediatric dermatology clinic, the parents of 44 young children with atopic eczema and 18 with other skin conditions were interviewed regarding sleep patterns. Sleep disorders were more prevalent in the eczema children than in the control group and normal populations. "Night waking" problems were particularly common in the eczema children, whereas settling problems were not. An association was found between scratching habits and "night waking"--the more the children scratched, the greater the likelihood of their waking at night. Consideration is given to the management of the sleep problems of eczematous children. PMID- 9394979 TI - Topical application of a platelet-activating factor (PAF) antagonist in atopic dermatitis. AB - Platelet-activating factor (PAF acether) is a lipid mediator with a potent proinflammatory activity. Results derived both from in vitro and in vivo studies suggest a possible role of this substance in the pathophysiology of atopic dermatitis. A double-blind, randomized, multi-center, within-patient study was performed to evaluate the efficacy of a topically applied PAF antagonist (RO-24 0238) in 36 patients with atopic dermatitis. Over a period of 28 days, 0.25 ml of the PAF antagonist and the vehicle (placebo) were applied twice daily on opposite sites of symmetrical lesions (measuring 10 to 20 cm2 each). The overall assessment of the therapeutic efficacy did not demonstrate a superior effect of the PAF antagonist in comparison to placebo, and this was the same with the individual study parameters erythema, scaling, induration and exudation. For reducing pruritus, as assessed by the patient using a visual analogue scale, a statistically significant action was documented during the first 2 weeks of the study (p < 0.04; Wilcoxon rank sum test), with a continued, yet not statistically significant efficacy after weeks 3 and 4. The exact role of the pathological events of atopic dermatitis that might be influenced by a PAF antagonist remains to be determined, but the anti-pruritic component of this substance especially deserves further scientific interest. PMID- 9394980 TI - Treatment of refractory cases of atopic dermatitis with acidic hot-spring bathing. AB - The incidence of refractory atopic dermatitis has increased in teenagers and young adults. The purpose of this study was to control the skin symptoms of such patients in daily life. Seventy patients repeatedly took a 10-min 42 degrees C acidic hot-spring bath twice daily. The skin symptoms were improved in 76% of cases. In 30 of 42 responders examined Staphylococcus aureus, detected on the skin surface, disappeared or decreased through balneotherapy. In contrast, S. aureus remained unchanged in 8 of 10 non-responders examined. Thus, the balneotherapy using acidic hot-spring water may be useful for controlling the skin symptoms of acute flares of refractory cases of atopic dermatitis. PMID- 9394981 TI - Evaluation of a self-reported questionnaire on hand dermatosis in secondary school children. AB - The purpose of this study was to investigate the reliability and validity of a self-reported questionnaire for estimating the prevalence of hand eczema and other hand dermatoses. All pupils in grades 1 and 3 from the secondary schools in Vaxjo, in southern Sweden, were invited to participate in the study, which consisted of two parts, a questionnaire and a clinical examination. Of those invited, 2572 (98.6%) responded to the questionnaire. Of the respondents, 2535 pupils (98.5%) were clinically examined. The kappa value for the questionnaire findings, compared with the diagnosis from the clinical examination, was 0.79, indicating good agreement. The sensitivity of the questionnaire findings was 73% (95% confidence interval [CI]: 0.6425-0.7975), and the specificity was 99% (95% confidence interval [CI]: 0.9860-0.9940). The self-reported questionnaire was suitable for detecting hand dermatosis in a population of secondary school children and may be used as a cost-effective and reliable method of investigating the prevalence of hand dermatosis in epidemiological studies. PMID- 9394982 TI - Plaque-type blue nevus. Review and an unusual Case. AB - A 31-year-old male presented with a 13-15-mm blue-black plaque tumor on his left forearm, which had existed since birth. Histological examination showed a common blue nevus. Its small size and its location are unusual. A review of plaque-type blue nevus is given. PMID- 9394983 TI - Improvement of vitiligo after oral treatment with vitamin B12 and folic acid and the importance of sun exposure. AB - The aim of this 2-year study was to test the hypothesis that folic acid, vitamin B12 and sun exposure could be helpful in treating vitiligo. One hundred patients with vitiligo were treated with oral folic acid and vitamin B12 after being informed that sun exposure might enhance repigmentation. They were requested to keep a record of sun exposure in summer and UVB irradiation in winter. The minimal treatment time suggested was 3-6 months but should be longer if improvement was achieved. Clear repigmentation occurred in 52 patients, including 37 who exposed their skin to summer sun and 6 who used UVB lamps in winter. Repigmentation was most evident on sun-exposed areas, where 38% of the patients had previously noted repigmentation during summer months. Total repigmentation was seen in 6 patients. The spread of vitiligo stopped in 64% of the patients after treatment. Folic acid and vitamin B12 supplementation combined with sun exposure can induce repigmentation better than either the vitamins or sun exposure alone. Treatment should continue as long as the white areas continue to repigment. Further studies are needed to determine ideal minimal dosages of vitamins and UV exposure, as well as treatment time. PMID- 9394984 TI - Epidermal sheet grafts for repigmentation of vitiligo and piebaldism, with a review of surgical techniques. AB - Thin epidermal sheets, obtained by a high-speed air-driven dermatome, were used to repigment white areas in 19 patients with vitiligo and one boy with piebaldism. In the depigmented skin to be treated the epidermis was removed by a rotating diamond fraise under topical and/or local anaesthesia injections. The method was used on most parts of the body, including the eyelids and genitalia. The maximum total area treated on each occasion was 190 cm2. Excellent results could be obtained if the vitiligo had been stable and had not increased anywhere during the last 2 years. Lack of immobilization could explain a poor result in some areas. The donor area on the buttocks healed quickly without depigmentation. In the transplanted area milia were observed in the first 6 months. No scarring was seen. The technique has a niche in the treatment of depigmented skin, especially in larger areas. PMID- 9394985 TI - Factors influencing participation among melanoma screening attenders. AB - We surveyed the demographic profile and motives prompting to participate among people attending voluntary melanoma screening clinics in Southern Limburg, the Netherlands, in June 1993. Precampaign public announcements addressed only melanoma and its precursor lesions. All attendees completed a detailed questionnaire addressing demographic particulars and specific fixed choice questions on their motivation to attend. There were 4,146 persons attending the screening clinics. Most attendees opted for examination of a specific lesion (71%). More females than males participated. Fear of having skin cancer was an important reason to participate (27%). Of all attenders, 16% had to be convinced by relatives or friends to attend the screens, and 33% would not have visited a physician on their own initiative when there had not been a free screening. Females were more concerned about skin cancer than males. The local and regional newspapers formed the most important precampaign publicity channel. Free melanoma screenings attract large numbers of people. Males are underrepresented. They are less aware of the risk profile of melanoma. Future screenings should target the male population. PMID- 9394986 TI - Pinch grafting of leg ulcers. A retrospective study of 412 treated ulcers in 146 patients. AB - In a retrospective study of 412 leg ulcers in 146 patients treated with pinch grafting, with a mean duration of follow-up of 32 months (range 2-84), the overall healing rate was 38%. The healing rate was best in the vasculitic ulcers (56%), followed by venous ulcers (38%), arteriosclerotic ulcers (33%), mixed ulcers (33%) and "other ulcers" (20%). In the series as a whole, the mean duration of ulcer problems was 8 years, and that of the 412 ulcers treated 2.5 years; the mean recurrence rate was 28%, and the mean remission time 12.5 months. In the ulcers that were still healed at close of the study (comprising 27% (112/412) of the series), the remission time was > or = 26.6 months. Thus we consider pinch grafting to be a successful complement to conservative therapy in most types of ulcers. PMID- 9394988 TI - Widespread cutaneous cryptococcosis occurring in an immunocompromised patient treated with high doses of fluconazole for oro-pharyngeal candidosis. PMID- 9394987 TI - Morphology of endogenous flare-up reactions in contact allergy to gold. AB - In a double-blind study, 20 patients with contact allergy to gold were given an intramuscular injection of gold sodium thiomalate or placebo, inducing a clinical and histological flare-up of healed patch test sites in the gold-injected but not in the placebo group. The test area of the placebo group showed some perivascular lymphocytic foci (UCHL-1+) and vascular endothelial ELAM-1 staining. The gold group, with flare-up, showed larger and more extensive lymphocytic foci with ELAM 1+ endothelium as well as lymphocytic epidermotropism. CD1a+ LC cells were downgraded, tryptase+ mast cells accumulated and CD68+ monocytes/macrophages markedly increased. Probably, a significant part of the tissue priming as a result of patch testing comprises memory T-cells and endothelial ELAM-1 upgrading, but blood-borne CD68+ monocytes may also be instrumental in the flare up. PMID- 9394989 TI - Koebner phenomenon in classic Kaposi's sarcoma. PMID- 9394990 TI - A pleomorphic liposarcoma imitated a subcutaneous cyst. PMID- 9394991 TI - Lithium therapy associated with hidradenitis suppurativa. PMID- 9394992 TI - Erythema multiforme major in a female with acute systemic meningococcal disease. PMID- 9394993 TI - Linear atrophoderma of Moulin. PMID- 9394994 TI - Acquired perforating collagenosis in a patient with carcinoma of the prostate. PMID- 9394995 TI - Cutaneous larva migrans detected by epiluminescent microscopy. PMID- 9394996 TI - Lichen planus and Crohn's disease. PMID- 9394997 TI - Epidermal growth factor concentration in the sera of male psoriatic patients. PMID- 9394998 TI - A case of dermatomyositis triggered by tegafur. PMID- 9394999 TI - Zosteriform lichen planus without evidence of herpes simplex virus or varicella zoster virus by polymerase chain reaction. Report of two cases. PMID- 9395000 TI - Lupus erythematosus as an occupational disease. PMID- 9395001 TI - Malignant epithelioid schwannoma with melanocytic differentiation: a rare tumour with an unusual feature. PMID- 9395003 TI - CO2 laser treatment causes local tattoo allergic reaction to become generalized. PMID- 9395002 TI - Low doses of low molecular weight heparin in vivo do not inhibit delayed-type hypersensitivity. PMID- 9395004 TI - The role of leukotriene A4 hydrolase/aminopeptidase in transcellular leukotriene B4 synthesis in human epidermis. PMID- 9395006 TI - Respiratory allergy to Cupressus sempervirens in Rome. AB - Mediterranean Cypress pollen is the major aerospore component in winter and early spring. Several recent studies have assessed the incidence of respiratory allergy to this pollen. A personal series of patients encountered in 1994-96 revealed a 9.33% incidence of positive prick-test responses to Cypress pollen among a population with atopical status. That series included 16 (19.05%) single and 68 (80.95%) multiple allergy sufferers. Among the former the symptoms encountered were rhinitis (62.5%) and asthma (37.5%). Given the ever-increasing incidence of Cypress pollen allergy, there is a need to restrict the planting of the tree for ornamental purposes, especially in areas with a high pollen count. PMID- 9395005 TI - Clinical efficacy and tolerance of two year Lolium perenne sublingual immunotherapy. AB - Last years, in spite of increasing use of sublingual immunotherapy (SLIT), not enough clinical studies have been published and its efficacy ought to be documented still more. For that purpose, 54 patients suffering seasonal rhinoconjunctivitis--with or without asthma--were allocated to either six month preseasonal SLIT with Lolium perenne extract (n = 35) or to a control group (n = 19). In the following year, thirty from previously treated patients and 12 from former control group, received a nine-month pre-coseasonal SLIT. Skin (SPT) and conjunctival (CPT) allergen response were monitorized several times along the study. Either seric antibodies or intraseasonal eosinophil cationic protein (ECP), as symptom, medication scores and peak nasal inspiratory flow (PNIF) were also assessed, fifty-five adverse reactions were recorded (0.7% doses), although only four required treatment. No main changes in CPT, SPT and antibodies were detected. Nevertheless, during the first pollen season, treated patients needed less medication than their control counterparts (p < 0.05). In the second season, the twelve ex-control subjects also required fewer drugs than in the first one (p < 0.01). Moreover, the whole forty-two treated patients showed a lesser intraseasonal ECP than a reference set of grass-allergic individuals (p < 0.05). We conclude that Lolium perenne SLIT is well tolerated and induces fewer drug requirements during pollen season, being eosinophil activation additionally reduced. PMID- 9395007 TI - Lodoxamide versus spaglumic acid: a comparative double-blind trial on patients suffering from seasonal allergic conjunctivitis induced by Parietaria pollen. AB - In order to evaluate the efficacy and tolerability of lodoxamide in the treatment of allergic conjunctivitis, the authors conducted a double-blind trial with intrapatient comparison on 32 patients, using lodoxamide versus spaglumic acid in the course of two conjunctival provocation tests performed with specific allergens. The patients received one drop of lodoxamide in one eye and one drop of spaglumic acid in the other; 15 minutes later, 25 microliters of allergen extract at a pre-established concentration was instilled. After 10 minutes, the signs and symptoms of the allergic response were evaluated and recorded. Six hours later, the instillation of the allergen extract in both eyes was repeated following the same procedure, to establish the duration of the effect of the two drugs. The results, obtained by evaluating the main clinical signs and symptoms (itching, lacrimation, hyperaemia, palpebral oedema and chemosis), demonstrate with statistically significant differences that lodoxamide inhibits the conjunctival response to exposure to the allergen with greater efficacy than spaglumic acid, and for a longer duration. The two drugs provided similar and satisfactory tolerability. In view of these results, lodoxamide can definitely be considered and effective drug in the treatment of allergic conjunctivitis. PMID- 9395008 TI - Modulation of TCR usage in HIV-1 infection is regulated by IL-7 and sCD23. AB - The purpose of this study is to elucidate the effect of interleukin-7 (IL-7) and soluble CD23 (sCD23) on Phorbol12 Myristate13 Acetate (PMA) activated CD4+ TCR alpha beta+ cells of HIV-1 infected subjects. CD23 and IL-7R were detectable on activated CD4+ T cells of these subjects by FACS. Addition on IL-7 (1000 U/ml), at the onset of cultures, resulted in a significant increase of CD23 expression. We also demonstrated that T cells proliferation and CD23 expression in the presence of exogenous IL-7 occur in an IL-2 independent manner. Addition of IL-7 and sCD23 to activated CD4+ cells of HIV-1 infected subjects induced a proliferative response of TCR alpha beta cells. In contrast, addition of either sCD23 or IL-7 to activated CD4+ T cells did not result in an increase of TCR alpha beta expression. The data provide direct evidence that sCD23 in combination with IL-7 induce proliferation of activated CD4+ T cells of HIV-1 infected subjects to augment TCR alpha beta expression. These results support the possibility that IL-7 plus sCD23 might play an important role in the modulation of TCR alpha beta expression in HIV infection. PMID- 9395009 TI - Fixed drug eruption from amoxycillin. AB - We present a case of fixed drug eruption on the genital mucosa induced by amoxycillin. Topical provocation was carried out, applying amoxycilin (10% pet) on the glans penis. No reaction was observed. Oral challenge with amoxycillin was followed by pruritic erythema on the glans penis 6 hours after the intake of 125 mg. The study of cross-reactivity to other betalactams showed good tolerance of phenoxymethyl-penicillin, which shares an identical nuclear structure with amoxycillin. The patient also tolerated cephadroxil, a cephalosporin with a side chain identical to that of amoxycillin. On reviewing the literature we have only found three reports of fixed drug eruption fue to amoxycillin. PMID- 9395010 TI - Platelet-activating factor antagonists. AB - Platelet-activating factor (PAF), identified as 1-O-alkyl-2-acetyl-sn-glyceryl-3 phosphorylcholine, exhibits potent proinflammatory properties. PAF is produced by numerous cell types, including endothelial cells, neutrophils, monocytes, macrophages, basophils, eosinophils and mastocytes. Since the discovery and identification of the chemical structure of PAF, a large variety of specific PAF receptor antagonists, both natural and synthetic compounds, have been described. Intensive research has been conducted and development programs set up by more 25 pharmaceutical companies world-wide, studying the therapeutic interest of more than 50 PAF-receptors antagonists in various pathophysiological conditions. Medline (1966-1996), Embase (Excerpta Medica; 1974-1996), and other biomedical and drug directory databases were searched to identify English-language articles (basic science, clinical trial research, and review articles) and abstracts of conference proceedings on PAF receptor antagonists and related terms. The most important PAF receptor antagonists are reviewed with their effectiveness in various experimental tests. Fundamentally, PAF antagonists may be divided in two groups: natural and synthetic compounds. Natural (Ginkgolides, Kadsurenone, Chantancin, Phomactin, Swietemohonin A, Prehispalone, THC-7-oic acid, Aglafoline, FR 900452, PCA 4248 and SCH 37370), and synthetic antagonists (CV-3988, CV-6209, SRI 63-072, SRI 63-441, UR-10324, UR-11353, E-5880, CL 184005, 6-Mono and Bis aryl phosphate antagonists, TCV-309, Ro-74719, WEB 2086, Y 24180, BN 50726, BN 50727, BN 50730, BN 50739, Ro 24-4736, Ro 24-0238, RP 55778, RP 59227, RP 66681, YM 264, YM 461, SM 10661, SR 27417, UK 74505, BB 182, BB 823, BB 654, SDZ 64-412, SDZ 65-123, L 652731, L 659898, L 668750, L 671284, L680573, L 680574, CIS 19, ABT-299 and Pinusolide) have a great variability in their chemical structure that might have importance in their different pharmacological profile. The great majority of these drugs are under development, and only a few have undergone clinical trials. PMID- 9395011 TI - Noncutaneous cavernous hemangiomas of the head and neck. PMID- 9395013 TI - Laryngeal reconstruction with free nonvascularized grafts from the buccal mucosa. AB - PURPOSE: This study was designed to investigate the suitability of nonvascularized healthy buccal mucosal graft in reconstruction of subglottic defects. PATIENTS AND METHODS: The study was conducted over an 11-year period from 1985 to 1995. Seven nonvascularized healthy buccal mucosal grafts were used to provide coverage for subglottic defects. The indications for surgery included chondrosarcoma of the cricoid including two recurrent tumors and one patient with recurrent subglottic benign laryngeal stenosis. No stenting was used. Period of follow-up ranged from 1 to 10 years. RESULTS: Graft take was successful in all cases without complications. CONCLUSION: Healthy nonvascularized buccal mucosal graft is suitable for reconstruction of subglottic defects. Stenting is not required. PMID- 9395012 TI - Using polymerase chain reaction to human papillomavirus in oral and pharyngolaryngeal carcinomas. AB - PURPOSE: Increasingly, evidence has shown that human papillomavirus (HPV) plays a role in the induction of certain carcinomas. The presence of HPV sequences in 56 previously untreated oral and pharyngolaryngeal carcinomas was examined by the polymerase chain reaction (PCR). MATERIALS AND METHODS: After DNA extraction, samples underwent 40 replication cycles with specific oligonucleotide primers corresponding to sequences from the E6 open-reading frame of HPV-6b, HPV-16, and HPV-18. To determine the E6 genomic integration, positive samples were processed with specific primers for the corresponding HPV L1 genes. Genomic HPV DNA clones into PBR 322 was used as positive control. RESULTS: HPV E6 DNA of the 6b and 16 types was detected in 14 patients (25%). The L1 gene was not present. CONCLUSION: Detected HPV E6 DNA might be integrated into the cell genome in the positive cases as indicated by the absence of the L1 gene-coding for the viral capside. Histological and survival rates, were unrelated to the presence of HPV. PMID- 9395015 TI - Surgical treatment for pulmonary metastases of squamous cell carcinoma of the head and neck. AB - PURPOSE: As locoregional control of head and neck cancer has improved, distant metastases have become increasingly common problems. PATIENTS AND METHODS: To determine the role of surgical treatment, we reviewed 32 patients with squamous cell carcinoma (SCC) of the head and neck who underwent thoracotomy for pulmonary metastases. RESULTS: The overall 5-year survival rate was 32%. The 5-year survival rate of the patients with SCC of the oral cavity was significantly poorer than that of the patients with other primary site (15.4% v 45.2%; P = .01). In the patients with single nodule, extent of the tumor was a significant prognostic factor (P = .007). Mediastinal lymph node involvement (P = .004) and pleural invasion (P = .04) also correlated with survival. CONCLUSION: TNM classification of the primary tumor did not have an impact on survival in this study. Further studies of a large series should be performed to determine the indications and modalities of the surgical treatment for pulmonary metastases of the SCC of head and neck. PMID- 9395014 TI - Supracricoid partial laryngectomy with cricohyoidoepiglottopexy for "early" glottic carcinoma classified as T1-T2N0 invading the anterior commissure. AB - PURPOSE: "Early" glottic squamous cell carcinoma classified as T1-T2N0 with anterior commissure invasion is conventionnaly managed with vertical partial laryngectomy (VPL) or radiation therapy (RT). At our insitution, in the early 1980s, vertical partial laryngectomy was progressively replaced by supracricoid partial laryngectomy with cricohyoidoepiglottopexy (SCPL-CHEP). The medical files and operative charts of 62 patients with "early" glottic carcinoma classified as T1-T2N0 invading the anterior commissure, consecutively managed with cricohyoidoepiglottopexy, were retrospectively reviewed to ascertain whether any conclusions could be drawn regarding this treatment modality. MATERIALS AND METHODS: Survival, local control, nodal recurrence, distant metastasis, and metachronous second primary tumor estimate was analyzed using the Kaplan-Meier life table method. RESULTS: The 3- and 5-year actuarial survival estimate was 93.3% and 86.5%, respectively. The 3- and 5-year actuarial local control estimate was 98.2%. The only patient with local recurrence was successfully salvaged with RT resulting in an overall 100% local control rate and laryngeal preservation rate. The 3- and 5-year actuarial nodal recurrence estimate was 1.8%. The 3- and 5-year actuarial distant metastasis estimate was 0% and 2%, respectively. Aspiration related completion total laryngectomy and permanent tracheostomy never occurred. CONCLUSION: The present retrospective study suggests that cricohyoidoepiglottopexy for glottic carcinoma classified as T1-T2 invading the anterior commissure resulted in higher local control rates and overall laryngeal preservation rate when compared with historical series using either VPL or RT. Further series are warranted to confirm our results. PMID- 9395017 TI - Fine-needle aspiration of head and neck masses in children. AB - PURPOSE: Head and neck masses in children are common. Suspicious or persistent masses are referred to the otolaryngologist who is faced with the dilemma of deciding which ones require surgical excision. Fine-needle aspiration (FNA) in adults helps distinguish lesions requiring excision from those that do not. Few reports exist of its use in children PATIENTS AND METHODS: Between January 1991 and December 1994, 67 FNAs were performed on children, 29 of which (43%) were for head and neck masses. Based on the FNA findings, 16 patients underwent surgery. RESULTS: In 13 patients, the final pathology was consistent with the FNA findings: granulomatous diseases (3), branchial cysts (3), acute/chronic lymphadenitis (3), thyroglossal cyst, hemangioma, Hodgkin's lymphoma, and Castleman's disease (one each). There was one misdiagnosis, no false positives, and two nondiagnostic specimens. Based on the results of FNA, surgery was not performed in the remaining 10 patients. The cytology was: cervical lymphadenopathy (7), abscess formation (1), lymphangioma (1), and leukemia (1). CONCLUSION: We conclude that FNA in an extremely useful tool in the management of head and neck masses in children. It is very well-tolerated by children, and we did not encounter any complications. PMID- 9395016 TI - Update on intraoperative analysis of mandibular margins. AB - PURPOSE: Assessing the adequacy of the surgical resection during composite resection of carcinoma is limited by the ability to evaluate the bone margins. The standard pathologic evaluation of bone is by decalcification. PATIENTS AND METHODS: A method of analysis was devised based on histologically proven methods of cortical invasion and subsequent spread. Frozen-section analysis of cancellous bone was investigated as a rapid method of evaluating adequacy of the mandibular resection. This report is an update of previously published results and includes an increase in the sample size as well as the analysis of additional pathology. Subjects consisted of 66 patients undergoing full thickness mandibular resection with 30 cases of histologically proven mandibular invasion qualifying for evaluation. Results on frozen section were then compared to the permanent section analysis on the cancellous bone and to the decalcified specimen. RESULTS: Complete correlation was found between frozen and permanent section results. Frozen section analysis was able to correctly predict adequacy of resection in 60 of 61 margins. CONCLUSION: Based on these results, the oncologic surgeon can evaluate bone margins at the time of the resection and adjust the amount of excision required to eradicate the disease. PMID- 9395019 TI - Cortical activity of a patient with Usher's syndrome using a cochlear implant. PMID- 9395018 TI - Eosinophil expression of transforming growth factor-beta and its receptors in nasal polyposis: role of the cytokines in this disease process. AB - PURPOSE: Nasal polyposis (NP) is characterized by an increase in inflammatory processes including fibrosis. Because transforming growth factor beta (TGF-beta) has been proven to induce fibrosis, we hypothesize that TGF-beta and its receptors are present in NP and influence polyp development. MATERIALS AND METHODS: To test this hypothesis, we evaluated distribution (immunohistochemistry) of TGF-beta isoforms (TGF-beta 1, TGF-beta 2, and TGF-beta 3) and its receptors [(TGF-beta(RI) & TGF-beta(RII)] in NP from 36 NP patients and in five normal sinus tissue specimens obtained from septoplasty/inferior turbinectomy. Tissue levels of TGF-beta 1 and TGF-beta 2 levels were determined by enzyme-linked immunosorbent assay (ELISA) and protein content was determined by Bio Rad assay (Bio Rad, Richmond, CA). All tissue levels of TGF-beta were normalized and expressed as pg of TGF-beta per mg of total protein (pg/mg TP). RESULTS: Immunohistochemical studies showed eosinophils as the major cells positively staining for TGF-beta 1, TGF-beta 2, TGF-beta 3, TGF-beta(RI), and TGF beta. In fibrotic sections, increased staining of eosinophils, fibroblast, and mononuclear cells was found for all three isoforms and both receptors. Evaluation of tissue levels indicated mean levels of TGF-beta 1 in the NP were 11.64 +/- 22.12 pg/mg TP versus normal control mean 44.36 +/- 22.12 pg/mg TP.TGF-beta 2 mean levels were 11.46 +/- 23.73 pg/mg TP versus normal control mean of 2.03 +/- 1.13 pg/mg TP. NP showed decreased expression of TGF-beta 1 and enhanced expression of TGF-beta 2 isoforms with presence of their receptors. Higher levels of TGF-beta 2 correlated with an increase in previous polypectomies perhaps indicative of severity of disease (P < or = .0001). CONCLUSION: Our studies show the presence of the TGF-beta isoforms and receptors in NP tissue. The results support our hypothesis that the eosinophil continues to be a pivotal inflammatory cell in NP, a differential regulation may govern the activity of TGF-beta in NP, and hence, the TGF-beta family of cytokines and receptors likely play a key role in controlling NP formation. PMID- 9395020 TI - Endonasal endoscopic management in fibrous dysplasia of the paranasal sinuses. PMID- 9395021 TI - Maxillary sinus ossifying fibroma. PMID- 9395022 TI - Bone spur presenting as a submandibular mass following free fibula reconstruction of the mandible. PMID- 9395023 TI - Candida epiglottitis: clinical implications. PMID- 9395024 TI - Endoscopic cranionasal resection of anterior skull base tumor. PMID- 9395025 TI - Alcohol-related traffic fatalities involving children--United States, 1985-1996. AB - Motor-vehicle-related injuries are the leading cause of death for persons aged 1 24 years in the United States. Although the relation between alcohol use and motor-vehicle-related deaths involving teenagers is well established, understanding of the role of alcohol in such deaths among younger children is limited. To characterize the involvement of alcohol in motor-vehicle-related deaths of U.S. children aged <15 years during 1985-1996, CDC analyzed data from the Fatality Analysis Reporting System (FARS) of the National Highway Traffic Safety Administration (NHTSA). This report summarizes the results of that analysis, which indicate that approximately one fourth of all traffic deaths among children aged <15 years involved alcohol and that in nearly two thirds of passenger deaths involving a legally drunk driver, the child was in the car driven by the legally drunk driver. PMID- 9395026 TI - Abortion surveillance: preliminary analysis--United States, 1995. AB - For 1995, CDC received data about legal induced abortions from the 50 states, New York City, and the District of Columbia. This report presents preliminary data for 1995; final abortion data for 1995 will be published during spring 1998. PMID- 9395027 TI - Use of clinical preventive services by Medicare beneficiaries aged > or = 65 years--United States, 1995. AB - Delivery of clinical preventive services to older adults can reduce premature morbidity and mortality while preserving function and enhancing overall quality of life . Until recently, the use of such services has been low among older adults because Medicare coverage has not been extended to many preventive services (3). Medicare coverage now includes four clinical preventive services: a single lifetime pneumococcal polysaccharide vaccination (vaccine plus any required revaccination and administration) (since 1981); annual influenza vaccination (vaccine and administration) (since 1993); and forwomen, biennial mammography screening (since 1991) and Papanicolaou smear screening every 36 months (since 1990). To assess current state-specific levels of use of these services among Medicare beneficiaries, CDC and the Health Care Financing Administration (HCFA) analyzed data from the 1995 Behavioral Risk Factor Surveillance System (BRFSS). This report summarizes the findings of this analysis, which indicate that, despite Medicare coverage of these preventive services, many U.S. adults aged > or = 65 years did not receive such services in 1995, and state-specific use of these services varied substantially. PMID- 9395028 TI - Efforts to quit smoking among persons with a history of alcohol problems--Iowa, Kansas, and Nebraska, 1995-1996. AB - In 1991, approximately 13.8 million adults in the United States met diagnostic criteria for alcohol abuse, alcohol dependence, or both. In addition, at least 80% of persons in this group were likely to be daily tobacco smokers and, therefore, at increased risk for oral and pharyngeal cancers. In Minnesota, among adult smokers with a history of alcohol abuse during 1972-1983, the number of tobacco-related deaths was higher than the number of alcohol-related deaths. To assess rates of smoking cessation among adults with a history of alcohol problems, the University of Nebraska Medical Center conducted an intervention study with 1 year of follow-up during 1995-1996 in 12 residential alcohol treatment centers in Iowa, Kansas, and Nebraska. This report summarizes the findings, which suggest that a substantial proportion of adults recently treated for alcoholism attempted to quit smoking, even though actual quit rates were low. PMID- 9395029 TI - Alcohol involvement in fatal motor-vehicle crashes--United States, 1995-1996. AB - The table and figure on page 1155 compare alcohol involvement in fatal motor vehicle crashes for 1995 and 1996. A fatal crash is considered alcohol-related by the National Highway Traffic Safety Administration (NHTSA) if either a driver or non-occupant (e.g., pedestrian) had a blood alcohol concentration (BAC) of > or = 0.01 g/dL in a police-reported traffic crash. Because BACs are not available for all persons in fatal crashes, NHTSA estimates the number of alcohol-related traffic fatalities based on a discriminant analysis of information from all cases for which driver or nonoccupant BAC data are available. PMID- 9395030 TI - Long-term follow-up of patients with newly diagnosed acute myeloid leukemia treated at the University of Texas M.D. Anderson Cancer Center. AB - BACKGROUND: Chemotherapy is known to cure a small minority of patients with acute myeloid leukemia (AML). Less is known about the risk of such patients developing subsequent cancers or about their ability to return to work. METHODS: The authors analyzed outcomes among 1892 patients who received treatment for newly diagnosed AML at the University of Texas M. D. Anderson Cancer Center from 1965 to May 1995. RESULTS: Because failure rates declined to relatively low levels after a first or later complete remission of > or = 3 years' duration, such patients comprised a "potentially cured" cohort. The criterion for entry into this cohort was fulfilled by 215 patients (10.7%; 203 in first complete remission and 12 in second remission). At a median of 6.2 years after entry into the cohort (i.e., 9.2 years from complete remission), 163 patients (76%) remain alive and in complete remission. Approximately 9% and 5% of the 1892 patients have been in complete remission for > 5 years and > 10 years, respectively. The pretreatment prognostic importance of cytogenetics is still apparent even after 5 years in complete remission. On average, members of the potentially cured cohort were not observed to be at increased risk of subsequent invasive malignancies compared with a normal population. Furthermore, two-thirds of those in the potentially cured cohort who were working full time before diagnosis of AML claimed to have returned to full-time work. Of those not working, only 10% cited physical limitation as the reason. CONCLUSIONS: The major threat to the life and well being of the patient with AML is clearly the disease and not its treatment. PMID- 9395031 TI - Long-term follow-up of three randomized trials comparing idarubicin and daunorubicin as induction therapies for patients with untreated acute myeloid leukemia. AB - BACKGROUND: Most clinical trials for acute leukemia have reported results after 2 3 years of follow-up. Comparisons between the original data and longer-term follow-up data may be of interest, particularly with regard to promising new therapies. METHODS: In 1996, survival data were updated from three prospective, randomized comparisons of idarubicin and daunorubicin that began in 1984 and 1985. These were trials of the Memorial Sloan-Kettering Cancer Center (MSKCC), the U.S. Multicenter Study Group, and the Southeastern Cancer Study Group (SEG). The original results of these trials were reported in 1991 and 1992. RESULTS: The original results of the SEG trial demonstrated no significant difference between idarubicin and daunorubicin. The updated survival analysis showed similar results. The MSKCC trial revealed a significant advantage of idarubicin compared with daunorubicin in both the original and the updated analyses. The U.S. Multicenter trial found a significant difference favoring idarubicin in the original analysis, but the difference was not significant in the updated analysis. CONCLUSIONS: It is essential that the median length of follow-up be clearly stated in any clinical trial. When the results obtained with a particularly promising new drug or procedure are presented early in the course of study (within 1-2 years), the investigators should strongly consider a repeat evaluation after an additional 3-5 years of follow-up. PMID- 9395032 TI - Long-term survival of patients with acute myeloid leukemia: updated results from two trials evaluating postinduction chemotherapy. AB - BACKGROUND: Although the prospect of long-term disease free survival (LFS) after chemotherapy for acute myeloid leukemia (AML) is widely accepted, few studies have reported long-term survival data. The authors therefore updated results from a 1981 report on a study conducted by the University of Minnesota Masonic Cancer Center (UMMCC) and a 1989 report on a study conducted by the North American Marrow Transplant Group (NAMTG). METHODS: Minimum follow-up of 21.6 years for living patients was obtained for 26 patients who received weekly cytarabine and 6 thioguanine maintenance therapy after achieving complete remission (CR) in the UMMCC study. Minimum follow-up of 7.7 years was obtained on 87 patients treated with high dose cytarabine intensification in first remission in the NAMTG study. RESULTS: In the UMMCC study, the LFS rate was 28% and the overall survival rate was 15%. Nineteen percent of patients died in first CR at 1.3-12 years. Three patients remain alive in initial CR at >20 years. In the NAMTG study, the LFS rate was 49% and the overall survival rate was 45%. A total of 38 patients (44%) remain alive in initial CR at a median of 11.4 years after diagnosis. An additional patient is alive in second CR at 8.6 years after diagnosis. In both studies, relapses after 3 years were relatively uncommon (11-12%). CONCLUSIONS: Chemotherapy alone is curative in more than 40% of AML patients who achieve CR. Short-term, high dose cytarabine intensification appeared more efficacious, without increased toxicity, compared with low dose, prolonged cytarabine-based maintenance. However, for patients who cannot receive intensification, prolonged, low dose maintenance therapy is an acceptable alternative for achieving cure. A minimum follow-up of 3 years is a reasonable predictor of long-term survival and should be obtained in studies evaluating therapeutic outcome in cases of AML. PMID- 9395033 TI - Long-term survival of patients with acute myeloid leukemia: a third follow-up of the Fourth International Workshop on Chromosomes in Leukemia. AB - BACKGROUND: In 1982, the Fourth International Workshop on Chromosomes in Leukemia reviewed data prospectively collected on 716 patients with acute myeloid leukemia (AML) diagnosed between 1980 and 1982. The present study examined the extended follow-up on these patients. METHODS: The analyses included cytogenetic and clinical data, with a median follow-up of 14.7 years, from 54 patients with treatment-associated AML and 628 with de novo AML. Of these patients, 291 received induction therapy that would be considered standard by today's criteria; no patient received high-dose cytarabine (HiDAC) intensification. RESULTS: Among the patients with treatment-associated AML, the only long-term survivor in retrospect appears to have had de novo AML. Among the patients with de novo AML, achievement of complete remission and survival varied significantly based on cytogenetic classification among all 628 patients as well as among those who did and did not receive standard induction therapy. The remission rate and survival were significantly better with standard induction therapy for patients with t(15;17) and normal cytogenetics. Multivariate analyses showed that karyotype was an independent predictor of survival for all patients and those receiving standard induction therapy. Only 8.9% of patients were alive 5 years following diagnosis, but 5 years of continuous remission was synonymous with cure. Even among 5-year survivors who had suffered a previous relapse, 41% appeared to be cured. Survival among patients in continuous remission for > or = 10 years varied significantly by cytogenetic classification. In the absence of HiDAC intensification, no complete responders with t(8;21) and only 7% with normal cytogenetics survived continuously 10 years disease free. CONCLUSIONS: Cure of AML following specific therapies must be evaluated in the context of cytogenetics. A meta-analysis incorporating cytogenetic data is indicated for patients with > or = 10 years of follow-up. PMID- 9395034 TI - Long-term survival after chemotherapy for acute myeloid leukemia: the experience of the Southwest Oncology Group. AB - BACKGROUND: Reports on outcomes of chemotherapy trials in acute myeloid leukemia (AML) have rarely included results of long-term follow-up beyond 10 years. The authors therefore chose to review long-term follow-up data from 3 studies conducted by the Southwest Oncology Group (SWOG) between 1978 and 1990. METHODS: The analysis included data on 2083 patients enrolled in SWOG studies S7823, S8124, and S8600. The results were based on data available as of November 15, 1996. RESULTS: The probability of survival 8 years after entry was 9% in Study S7823, 14% in S8124, and 15% in S8600. For patients age < 50 years, the probabilities were 14%, 24%, and 20%, respectively. For patients ages 50-64 years, the probabilities were 7%, 8%, and 8%, respectively. For those age < 50 years who achieved complete remission, the 8-year probability of disease free survival was 17% in Study S7823, 28% in S8124, 17% with standard dose cytarabine in S8600, and 26% with high dose cytarabine in S8600. Relapse was the major reason for failure after complete remission in all three studies. When the results of the 3 studies were combined, most of the 743 relapses had occurred by Year 3 and nearly all the rest by Year 5. Among the prognostic factors universally available for study, three were highly associated with survival in all three studies: age, French-American-British disease classification, and white blood cell count at diagnosis. CONCLUSIONS: In view of the fact that most deaths occurred during the first 3 years, it is appropriate to report the results of clinical trials after patients have been followed for 4 years. Despite modest gains, the results of chemotherapy for AML remain disappointing, especially in the treatment of older patients. PMID- 9395036 TI - Long-term follow-up of Cancer and Leukemia Group B studies in acute myeloid leukemia. AB - BACKGROUND: During the past 3 decades, the Cancer and Leukemia Group B (CALGB) has conducted numerous large clinical trials in adult patients with acute myeloid leukemia. METHODS: Updated data were obtained for 9 trials initiated by the CALGB between 1974 and 1992. The updated data were compared with the original published results. RESULTS: It was apparent from all the studies that hazard rates for death and relapse are greatest in the first year, decrease substantially between Years 1 and 2, then decrease further between Years 2 and 3. Rates of death and relapse are quite low after 3-4 years. Large numbers of patients are long-term disease free survivors. Overall, these patients have excellent function and a normal quality of life. CONCLUSIONS: Patients with AML who are in complete remission for 3-4 years can be assured that late relapse and death are relatively uncommon events. It is inadvisable to publish results of large studies until this minimal level of follow-up has been reached. PMID- 9395035 TI - Long-term survival in acute myeloid leukemia: the Eastern Cooperative Oncology Group experience. AB - BACKGROUND: The data base of the Eastern Cooperative Oncology Group (ECOG) provides access to data on a large adult patient population drawn from more than 25 major university institutions and hundreds of participating hospitals. Extensive medical files are maintained at the ECOG Coordinating Center and are updated regularly. METHODS: Data on 1414 eligible patients with acute myeloid leukemia (AML), treated on 6 ECOG protocols during the period 1976-1994, were reviewed to determine the number of long-term survivors (LTS) and to identify factors that predicted LTS. Disease free survival and factors impacting quality of life were examined as well. RESULTS: Of the 1414 patients, 274 survived for > or = 3 years and were considered LTS. A logistic regression analysis revealed that factors predicting LTS included age < 55 years, female gender, treatment between 1985 and 1990, white blood cell count < 10,000 cells/mm3, and hemoglobin > 10 g/dL. Disease free survival improved with escalating intensity of therapy. Quality-of-life data showed that infections were fairly common. Significant graft versus-host disease occurred in 6 of 40 patients who received allogeneic bone marrow transplantation and contributed to the deaths of 4 individuals. Information on employment, insurance, social or marital difficulties, and psychosocial issues was more difficult to obtain. CONCLUSIONS: Prognosis in AML is a complex interaction involving the cellular origin of the malignant clone, morphology, and evolving therapeutic strategies. The most recent ECOG studies incorporate these variables and should provide additional insights into factors affecting LTS in patients with AML. PMID- 9395037 TI - Conclusions regarding leukemia long-term survival. PMID- 9395039 TI - Mechanical valve closing dynamics: relationship between velocity of closing, pressure transients, and cavitation initiation. AB - In this study, the closing dynamics of mechanical heart valves was experimentally analyzed with the valves mounted in the mitral position of an in vitro flow chamber simulating a single closing event. The average linear velocity of the edge of the leaflet during the final 2.065 degrees of the traverse before closing was measured using a laser sweeping technique, and the negative pressure transients at 2 mm from the leaflet inflow surface in the fully closed position was recorded at the instant of valve closure. The cavitation number was computed for the various mechanical valves at a range of load at valve closure. The data were correlated with cavitation bubble visualization previously obtained with the same experimental set up. Cavitation incipience with mechanical valves was found to be independent of the flexibility of the valve holder. For the same loading rate at valve closure, valves with flexible (polyethylene) leaflets were found to close with comparable velocity to those with rigid (pyrolytic carbon) leaflets, but the negative pressure transients did not reach magnitudes close to the vapor pressure for the fluid with flexible leaflets. For the same leaflet closing velocity (and hence the cavitation number), valves with a seat stop or a seating lip in the region of maximum leaflet velocity were observed to cavitate earlier, suggesting that the effect of "squeeze flow" may be an important factor in cavitation incipience. PMID- 9395038 TI - Thin layer flows due to surface tension gradients over a membrane undergoing nonuniform, periodic strain. AB - Measurements of surface tension in the lung have shown that a time-mean gradient exists with the potential to generate clearance flows toward the mouth in the thin liquid layer that lines the airways. A model is developed to explore this phenomenon in the simple case of a membrane with linear variation in strain along its length, coupled with the unique properties of pulmonary surfactant. The evolution equations are solved numerically for liquid layer thickness and surfactant concentration during a single oscillatory cycle, and the net volume exchanged is computed. The parameters governing the flow are shown to be time scales for viscous effects, tau(v), surface diffusion, tau(DS), surfactant adsorption, tau(A), surfactant desorption, tau(D), oscillation, tau(o), and the average membrane strain epsilon. The volume pumped toward the less compliant end on the initial cycle is maximized when tau(o)/tau(v) approximately O(1) and is relatively insensitive to tau(DS). Rapid adsorption generally augments liquid transport for tau(o)/tau(D) < O(1). Pumping drops precipitously if tau(o)/tau(D) > O(1). Effects of strain amplitude are reported as well. For parameter values approximating those in the lung, pumping rates are near optimal; the mean surface velocity is approximately 0.05 mm/sec, compared with 0.2 mm/sec produced by the action of cilia on the mucus layer. This mechanism might therefore be important in assisting clearance from the lung or maintaining a liquid layer over alveolar facets. PMID- 9395040 TI - Velocity measurements within confined turbulent jets: application to cardiovalvular regurgitation. AB - An expression for centerline mean velocity distributions for circular and noncircular confined turbulent jets has been obtained by assuming self preservation of flow downstream of the jet potential core. It was assumed that the velocity decay was not only dependent on the streamwise distance x in terms of x/d, as in the case of free jets, but also on the ratio of the orifice diameter d to the confining pipe diameter D. To validate the expression and to determine the empirical constants, measurements of the centerline velocities within the confined jets issuing from different size circular orifices and various noncircular orifices of different shapes were conducted. The results indicate that the validity of the expression is restricted to d/D < or =0.25 and is weakly dependent on the particular orifice shape. It is suggested that, as for the case of free turbulent jets reported earlier, that this expression may be used potentially to predict the valvular lesion size or to estimate the volume of valvular regurgitation for confined jets provided the value of D, which corresponds to the "atrial diameter," is known or statistically available. PMID- 9395041 TI - The role of spatial interactions in creating the dispersion of transmembrane potential by premature electric shocks. AB - Strong electric shocks applied during the refractory period can initiate or terminate cardiac arrhythmias. To elucidate the underlying mechanism, Knisley et al. used rabbit papillary muscle in vitro to scan the refractory period of an action potential with shocks of different strengths. The resulting map of the shock-induced changes in the transmembrane potential (Vm) illustrates the substrate for the creation of rotors. Our study uses computer simulations to reproduce this experimental map. Three models (a space-clamped membrane, a single cell, and a one-dimensional fiber) were used to determine whether the observed map was caused by (i) the intrinsic dynamics of the membrane, (ii) the simultaneous depolarization and hyperpolarization of the opposite ends of each cell, or (iii) spatial interactions involving the whole muscle strand. The results show that the membrane and single cell models cannot reproduce the experimental map. The fiber model reproduces the shock-induced changes in Vm and demonstrates that they are caused by a propagating disturbance, which, depending on the coupling interval and the shock strength, can be a new action potential or an electrotonus and can arrive from the depolarized end or from both depolarized and hyperpolarized ends of the fiber. These results indicate that the induction of rotors in the heart may not be a direct effect of the electric field. PMID- 9395043 TI - Time-frequency coherence analysis of atrial fibrillation termination during procainamide administration. AB - A time-frequency coherence estimator is developed and applied to study changes in signal characteristics as atrial fibrillation converts to sinus rhythm during administration of procainamide. A coherence spectrogram (CS) using multiple sinusoidal tapers is used in this study to assess phase relations between electrogram recordings at multiple atrial sites of seven patients who received procainamide to terminate atrial fibrillation. CSs are calculated (0 to 60 Hz) with 1 sec time resolution and 6.2 Hz frequency resolution. In agreement with previous studies, CSs generally exhibit low coherence during atrial fibrillation. Conversion to sinus rhythm is concomitant with an increase in coherence and emergence of structured time-frequency topography. Transition from atrial fibrillation to sinus rhythm is associated with a variety of time-frequency dynamics. Both gradual and abrupt increases in coherence coincide with conversion. Results suggest transient electrical organization in the atria during atrial fibrillation not seen in previous low-resolution coherence studies. CSs permit investigation of rhythm organization with unparalleled time and frequency resolution and thus are useful for studying transient changes in cardiac rhythms that may reflect underlying mechanisms. PMID- 9395042 TI - Mechanisms by which thrombolytic therapy results in nonuniform lysis and residual thrombus after reperfusion. AB - A transport-reaction model describing penetration of plasmin by diffusion and permeation into a dissolving fibrin gel was solved numerically to explore mechanisms that lead to the formation and growth of dissolution fingers through blood clots during thrombolytic therapy. Under conditions of fluid permeation driven by arterial pressures, small random spatial variations in the initial fibrin density within clots (+/-4 to 25% peak variations) were predicted by the simulation to result in dramatic dissolution fingers that grew in time. With in vitro experiments, video microscopy revealed that the shape of the proximal face of a fibrin gel, when deformed by pressure-driven permeation, led to lytic breakthrough in the center of the clot, consistent with model predictions of increased velocities in this region leading to cannulation. Computer simulation of lysis of fibrin retracted by platelets (where more permeable regions are expected in the middle of the clot due to retraction) predicted cannulation of the clot during thrombolysis. This residual, annular thrombus was predicted to lyse more slowly, because radial pressure gradients to drive inner clot permeation were quite small. In conjunction with kinetic models of systemic pharmacodynamics and plasminogen activation biochemistry, a two-dimensional transport-reaction model can facilitate the prediction of the time and causes of clot cannulation, poor reperfusion, and embolism during thrombolysis. PMID- 9395044 TI - A comprehensive simulator of the human respiratory system: validation with experimental and simulated data. AB - A comprehensive model of oxygen (O2) and carbon dioxide (CO2) exchange, transport, and storage in the adult human is presented, and its ability to provide realistic responses under different physiological conditions is evaluated. The model comprises three compartments (i.e., lung, body tissue, and brain tissue) and incorporates a controller that adjusts alveolar ventilation and cardiac output dynamically integrating stimuli coming from peripheral and central chemoreceptors. A new realistic CO2 dissociation curve based on a two-buffer model of acid-base chemical regulation is included. In addition, the model explicitly considers relevant physiological factors such as buffer base, the nonlinear interaction between the O2 and CO2 chemoreceptor responses, pulmonary shunt, dead space, variable time delays, and Bohr and Haldane effects. Model simulations provide results consistent with both dynamic and steady-state responses measured in subjects undergoing inhalation of high CO2 (hypercapnia) or low O2 (hypoxia) and subsequent recovery. An analysis of the results indicates that the proposed model fits the experimental data of ventilation and gas partial pressures as some meaningful simulators now available and in a very large range of gas intake fractions. Moreover, it also provides values of blood concentrations of CO2, HCO3-, and hydrogen ions in good agreement with more complex simulators characterized by an implicit formulation of the CO2 dissociation curve. In the experimental conditions analyzed, the model seems to represent a single theoretical framework able to appropriately describe the different phenomena involved in the control of respiration. PMID- 9395045 TI - Nonparametric block-structured modeling of rat lung mechanics. AB - The quasistatic and dynamic pressure volume characteristics of the lungs were measured in five anesthetized, paralyzed open-chest rats. Psuedo-random volume perturbations over a frequency range of 0.25 to 25 Hz and having peak-peak amplitudes of 1 to 4 ml were applied after the lungs were allowed to expire against 0.2, 0.4, 0.6, and 0.8 kPa positive end-expiratory pressure (PEEP). The lung mechanics were partitioned in two ways: a linear dynamic block followed by a static nonlinearity (Wiener model) and a static nonlinearity ahead of a linear dynamic block (Hammerstein model). It was found that a Hammerstein model featuring a third-order polynomial static nonlinearity and a linear impulse response function of 1-sec duration accounted for the greatest amount of the output variance (98.8 +/- 0.6%, mean +/- SD from perturbations of 4 ml amplitude and PEEP = 0.8 kPa). The static nonlinear behavior matched the measured quasistatic pressure volume behavior obtained at the same amplitude and at the same level of PEEP, provided that all direct current gain of the model was located within the static nonlinearity. Under these conditions, the linear resistance was inversely dependent on the PEEP, whereas little PEEP or amplitude dependence of the linear compartment elastance was observed. Thus, of the two block-structured models tested, the Hammerstein model accounted better for the large amplitude nonlinear mechanical behavior. However, neither model could account for the dependence of the linear block resistance on PEEP. PMID- 9395046 TI - Experimental and numerical analyses of indentation in finite-sized isotropic and anisotropic rubber-like materials. AB - Indentation tests perpendicular to the major plane of a material have been proposed as a means to index some of its in-plane mechanical properties. We showed the feasibility of such tests in myocardial tissue and established its theoretical basis with a formulation of small indentation superimposed on a finitely stretched half-space of isotropic materials. The purpose of this study is to better understand the mechanics of indentation with respect to the relative effects of indenter size, indentation depth, and specimen size, as well as the effects of material properties. Accordingly, we performed indentation tests on slabs of silicone rubber fabricated with both isotropic, as well as transversely isotropic, material symmetry. We performed indentation tests in different thickness specimens with varying sizes of indenters, amounts of indentation, and amounts of in-plane stretch. We used finite-element method simulations to supplement the experimental data. The combined experimental and modeling data provide the following useful guidelines for future indentation tests in finite size specimens: (i) to avoid artifacts from boundary effects, the in-plane specimen dimensions should be at least 15 times the indenter size; (ii) to avoid nonlinearities associated with finite-thickness effects, the thickness-to-radius ratio should be >10 and thickness to indentation depth ratio should be >5; and (iii) we also showed that combined indentation and in-plane stretch could distinguish the stiffer direction of a transversely isotropic material. PMID- 9395047 TI - Extraction forces and tissue changes during explant of CWRU-type intramuscular electrodes from rat gastrocnemius. AB - Intramuscular electrodes are currently in use for clinically implementing several electrotherapeutic and neuroprosthetic protocols. A decrease in motor recruitment is often reported in these systems due to movement of the electrode tip from the initial implant site. In the study reported here, multistrand intramuscular electrodes of the CWRU design were implanted aseptically in the gastrocnemii of adult rats under anesthesia. These electrodes were explanted immediately after implant in one group and after periods of 1 and 4 hr; 1, 3, and 5 days; 1 week; 10 days; and 2 and 4 weeks in others. Force as a function of displacement was recorded during explantation. Analysis of the results showed that there was a significant increase in the force required to dislodge the electrode tip between 5 and 7 days of implant. Electrodes seemed to be vulnerable to movement in the first 5 days when the barb provided the only fixation. Histology of muscles from which electrodes had been explanted did not show any increase in the area of tissue changes, compared with control muscles in which the electrode remained in situ. These results indicated that electrode removal occurred within the encapsulation tissues, and the surrounding muscle was mainly unaffected by the explant process. PMID- 9395048 TI - Segmentation of depth-EEG seizure signals: method based on a physiological parameter and comparative study. AB - The analysis of stereoelectroencephalographic (intracerebral recording) signals provides information on the electrical activity of brain structures implied in epileptic seizures. A simple nonparametric adaptive segmentation method, based on a physiologically relevant parameter, is presented and compared with three methods reported in the literature. The comparative frame allows us to objectively test methods for their performances on the same basis. Results show that the proposed method is robust with respect to the types of change studied and easier to conduct, even if it is less accurate about the estimation of instants of change than another method presented in this study. Signals are segmented throughout the duration of seizures without parameter readjustment and generate instants of change in accordance with those interactively delimited by the clinician. PMID- 9395049 TI - An artificial neural network-based controller for the control of induced paralysis using vecuronium bromide. AB - This study presents an artificial neural network-based controller for regulating the level of induced paralysis during surgery using vecuronium bromide. The controller uses the myogram of a rapid muscle contractions (called twitch) to generate the appropriate infusion rate. The controller is self-adjusting and can accommodate inter- and intrapatient drug response variations. It also withstands changes in the pure time delay and nonlinear pharmacokinetic parameters of the response. Another feature of the controller is that it does not depend on a priori knowledge of the patient response model. Computer simulations using pharmacokinetic and pharmacodynamic models showed negligible steady-state error and maximum percent undershoot averaged to 6.24%. The average infusion rate for 90% paralysis was 1.22 (microg x kg(-1) x min[-1]). PMID- 9395050 TI - Do increased electrogastrographic frequencies always correspond to internal tachygastria? AB - This study was undertaken to investigate the possible origin of some cutaneously recorded higher frequency electrogastrographic signals. Computer modeling of gastric electrical uncoupling was performed using previously described conoidal dipole model. Cutaneous electrogastrograms were simulated after uncoupling was introduced. In separate, real-life experiments, 6 pairs of bipolar electrodes were inserted into the gastric wall (3 anterior, 3 posterior) of 15 anesthetized dogs at laparotomy to record 6 channels of internal gastric electrical activity (GEA). Eight-channel bipolar cutaneous electrogastrography (EGG) was simultaneously recorded. Three separate 1/2-hr recordings were made from each dog in the basal state and after each of two circumferential cuts of all gastric muscle. Distal stomach was surgically divided into three equal-sized areas, each with an electrode pair in its anterior and posterior walls. Gastric electrical activity and EGG were digitized, bandpass-filtered and analyzed in frequency domain using the fast Hartley transform. The phenomena of tachygastria and tachyarrhythmia were quantitatively compared in internal and cutaneous recordings. Computer modeling indicated that it is possible to record cutaneous "tachygastric" or "tachyarrhythmic" signals without them being present internally. Real experiments on dogs showed higher percentage of tachyarrhythmia in EGG than in the internal signal in the basal state. After the first circumferential cut, the periods of tachygastria and tachyarrhythmia increased, with EGG signals showing again a higher percentage. This tendency persisted after the second circumferential cut. A similar pattern was observed when monitoring the percentage of uncorresponding tachygastrias/tachyarrhythmias. Our findings indicate that gastric electrical uncoupling can be another possible reason for abnormal frequency characteristics of EGG. Not all cutaneously recognized tachygastrias and/or tachyarrhythmias could be related to objectively existing internal tachygastric events. PMID- 9395051 TI - Automatic segmentation of intravascular ultrasound images: a texture-based approach. AB - Extraction of blood vessel boundaries from intravascular ultrasound images is essential in the quantitative analysis of cardiovascular functions. In this study, we are presenting a completely automated procedure for determining blood vessel borders. This approach uses textural operators to separate different tissue regions and morphological processing to refine extracted contours. The method was tested in a set of 29 intravascular ultrasound images obtained in vivo. To assess the performance of the method, we have compared the automatically processed images with the manual tracings, using three different criteria: correlation coefficient, match ratio, and relative error of computed shape parameters. In both contour detection and shape parameters estimation, the proposed method yielded consistently good results. Due to its robustness and accuracy, this approach is appropriate for clinical use, whereas computational efficiency of the method facilitates low-cost implementation. PMID- 9395052 TI - Controlling receptor-ligand contact to examine kinetics of T cell activation. AB - A method for controlling the contact of cell-surface receptors with immobilized ligands has been developed. Cells are trapped in an asymmetric liquid film that can be quantitatively thinned by reducing the film's capillary pressure. Ligands adsorbed to the liquid-solid interface are forced into increasingly tighter contact with the cells as the air-liquid interface is drawn down. Controlling the degree of thinning allows study of repulsive forces, and controlling its time course produces a definite time 0 for analyzing signal transduction. This system was tested by examining the time course of calcium mobilization in T cells upon activation with anti-CD3 antibody at different dilutions and ionic strengths. The averaged calcium transient of the responding cells was essentially the same for each condition. However, the fraction of responding cells decreased with anti-CD3 dilution, and indicated that the critical ligand density for T cell activation lies between approximately 35 and 70 molecules of anti-CD3 per microm2. Decreasing the medium's ionic strength from the normal value of 157 mM to 57 mM did not affect either the average calcium response profile or the fraction of responding cells, but strongly affected receptor-ligand contact, decreasing the percent of spontaneous activation from 38% to 5%. Such an imposed decrease sets the stage for film thinning to impose much greater control of receptor-ligand contact. PMID- 9395053 TI - Three-component laser Doppler velocimetry measurements in the regurgitant flow region of a Bjork-Shiley monostrut mitral valve. AB - Three-dimensional laser Doppler velocimetry measurements were acquired in a mock circulatory loop proximal to a Bjork-Shiley monostrut valve in the mitral position, and synchronous ensemble-averaging was applied to form an "average" beat. Two axial locations in the regurgitant flow region of the valve (in the minor orifice) were mapped, and maximum Reynolds shear stresses were calculated. A large spike in regurgitant flow was noted at the beginning of systole, which may be the squeeze flow phenomenon computed by other researchers. A region of sustained regurgitant flow 50 msec later was the focus of this study. Maximum velocities of approximately 3.7 mps were noted, and maximum Reynolds shear stresses of approximately 10,000 dyne/cm2 were calculated. Comparisons were made of two-dimensional (ignoring tangential component) versus three-dimensional shear stresses, and, in this case, in regions of high stress, the differences were insignificant. This suggests that the tangential component of velocity can probably be ignored in similar measurements where the tangential velocity is likely to be small. PMID- 9395054 TI - Advances and pitfalls in the diagnosis of Lyme disease. PMID- 9395055 TI - Aggregation-promoting factor in human vaginal Lactobacillus strains. AB - A total of 60 Lactobacillus sp. strains were examined for expression of auto aggregation and cell surface hydrophobicity. Isolates were obtained from the vagina of healthy women (n = 20). The results obtained showed that the occurrence of cell surface hydrophobicity correlated with auto-aggregative activity in 12 homofermentative Lactobacillus sp. strains. The aggregation mechanism was mediated by the presence of an aggregation promoting factor (APF) in one Lactobacillus sp. strain, HV 142. APF was confirmed by DNA hybridisation with a 1.3 kb PstI DNA fragment of recombinant plasmid pFDS containing the reading open frame of the APF gene derived from Lactobacillus plantarum 4B2. Coaggregation activity was seen in three strains of auto-aggregative human vaginal lactobacilli and five strains of P-fimbriated uropathogenic Escherichia coli. Moreover, one of four Lactobacillus sp. strains (HV 389) aggregated with two of five E. coli tested. These results suggest that APF producing lactobacilli could represent a further mechanism in the interaction of commensal microflora with strains of uropathogenic E. coli. PMID- 9395056 TI - Epitope mapping Candida albicans proteinase (SAP 2). AB - The continuous epitopes of Candida albicans proteinase SAP 2 were derived by epitope mapping with sera from patients with oral candidiasis (n = 3), necropsy proven disseminated candidiasis (n = 5), paired sera from patients who had recovered from blood culture-proven disseminated candidiasis (n = 3) and infection due to Candida parapsilosis (n = 2) and Candida tropicalis (n = 2). In C. albicans infection, IgM identified epitopes in amino acid positions 57-61 (QAVPV), 146-151 (SQGTLY) and 346-351 (PYDKCQ) and IgG at position 386-390 (VKYTS). For C. tropicalis IgM and IgG were positive for the same epitopes whilst IgG also detected epitopes at 78-83 (SNNQKL) and 159-164 (GVSIKN). For C. parapsilosis, IgM was positive for SNNQKL and IgG detected no epitopes. Reactivity of two of the epitopes as peptides KTSKRQAVPVTL and SLAQVKYTSASSI was confirmed in an indirect ELISA. At a cut-off optical density of 0.4, IgM against either peptide was associated with survival but present in only about half of the sera (n = 60) from patients who recovered from disseminated candidiasis whilst IgG levels were disappointing. Human recombinant antibodies from a patient who had recovered from disseminated candidiasis against either of these peptides had no activity in a lethal mouse model of candidal infection. PMID- 9395057 TI - Differentiation between Debaryomyces hansenii/Candida famata complex and Candida guilliermondii by polymerase chain reaction. AB - Primers for the differentiation of the Debaryomyces hansenii/Candida famata complex were designed from large subunit ribosomal DNA base sequences. With these primers, a polymerase chain reaction test amplified DNAs from all the species of the Debaryomyces hansenii/Candida famata complex and distinguished this complex from Candida guilliermondii. PMID- 9395058 TI - Enhanced production of inducible nitric oxide synthase by beta-glucans in mice. AB - We have already demonstrated that various activities including NO (nitric oxide) synthesis in vivo and in vitro significantly differ between triple helical (SPG) and single helical (alkaline-treated SPG, SPG-OH) beta-glucans. It was previously suggested that the single helical conformer of beta-glucan (SPG-OH) was dominant in cytokine production and subsequent NO synthesis in vitro. In this study, we analyzed production of inducible nitric oxide synthase (iNOS) induced by beta glucans in vitro and in vivo. The iNOS production was enhanced in proteose peptone-induced peritoneal macrophages (PMs) cultured with SPG-OH in the presence of IFN-gamma for 24 h, and SPG-OH-induced PMs. Moreover, SPG-OH was effective for iNOS production not only in isolated macrophages but also in tissue macrophages, whereas SPG was less effective. These findings suggest that a single helical conformer is essential for iNOS production, and that NO synthesis by beta-glucans is closely related to iNOS production. PMID- 9395059 TI - Cloning and characterization of a Campylobacter jejuni 72Dz/92 gene encoding a 30 kDa immunopositive protein, component of the ABC transport system; expression of the gene in avirulent Salmonella typhimurium. AB - Three gene libraries of Campylobacter jejuni 72Dz/92 DNA were prepared using lambda gt11, pSupercos and pWSK129 cloning vectors. Screening of the libraries with Escherichia coli absorbed antiserum generated against whole C. jejuni revealed several immunoreactive clones of apparent molecular masses 19, 28, 30 and 50 kDa. The most commonly isolated clones expressed 30 kDa protein. The nucleotide sequence of the 1768 bp C. jejuni DNA yielded one complete (ORF2) and two partial open reading frames (ORF1 and ORF3). ORF2 encoded CjaA protein exhibits relevant overall homology to several prokaryotic solute binding proteins (family 3), components of the ABC transport system, while the product of the truncated ORF3 (CjaB protein) shows extensive homology to Gram-negative bacterial proteins, members of the sugar transporter family. The genetic organization of the putative cjaAB operon was studied. The cjaA gene fragment (616 bp) was amplified from three C. jejuni strains isolated from patients with acute bloody diarrhea, whereas it was not amplified from strains which caused acute diarrhea with no blood in the stools. The gene was introduced into avirulent Salmonella typhimurium vaccine strain where it is expressed at a reasonably high level. PMID- 9395060 TI - Simultaneous detection of Streptococcus pneumoniae and Haemophilus influenzae by nested PCR amplification from cerebrospinal fluid samples. AB - Haemophilus influenzae and Streptococcus pneumoniae are often the cause of serious diseases such as meningitis. We designed a nested PCR assay to identify these pathogens from cerebrospinal fluid samples. The first-step PCR was able to detect eubacterial rRNA genes with a unified set of universal primers. In the second-step PCR, the identification primers, HI I and II and SP I and II, could detect H. influenzae and S. pneumoniae respectively through amplification of the rRNA spacer between the 16S and 23S rRNA genes. We suggest that the two-step PCR assay can be used as a novel method for the immediate and retrospective diagnosis of bacterial meningitis caused by H. influenzae and S. pneumoniae. PMID- 9395061 TI - Analysis of the human Ig isotype response to individual transferrin binding proteins A and B from Neisseria meningitidis. AB - Subcapsular antigens, including transferrin binding proteins, are being considered as potential vaccines against serogroup B meningococci. This study examined the human isotype antibody responses in cases of meningococcal disease to meningococcal TbpA (transferrin binding protein A) and TbpB (transferrin binding protein B) from two strains (SD and B16B6) expressing high and low molecular mass TbpB respectively. TbpA isolated from both strains were recognised more frequently and higher durable ELISA absorbance values were detected than those detected against TbpB from either strain. These antibody responses to Tbps were independent of the infecting meningococcal strain type. The antibody response to the four proteins was highly variable between individuals and differed significantly against all four antigens. The variability of immune responses to each Tbp from the two strains suggests that a successful vaccine would need to include TbpA and TbpB from a number of strains. PMID- 9395062 TI - Interleukin-4 suppresses antifungal activity of human mononuclear phagocytes against Candida albicans in association with decreased uptake of blastoconidia. AB - Pathogenesis of invasive candidiasis may involve regulatory activities of Th2 immunity on phagocytic host defenses. The effects of interleukin (IL)-4 on antifungal capacity of human mononuclear phagocytes against Candida albicans were studied. Incubation of adherent mononuclear leukocytes from healthy donors with IL-4 (1-5 ng ml(-1)) at 37 degrees C for 2-4 days suppressed uptake of C. albicans blastoconidia in the presence of human serum (P < or = 0.01), and anti IL-4 inhibited its suppressive effect. The effect of IL-4 was protein synthesis dependent. Interferon-gamma (0.25-25 ng ml(-1)), granulocyte-macrophage colony stimulating factor (CSF, 20 ng ml(-1)), macrophage-CSF (15 ng ml(-1)) but not IL 10 (100 ng ml(-1)) somewhat counteracted the suppressive effect of IL-4. In contrast, mannose receptor-mediated uptake of blastoconidia in the absence of serum was increased by IL-4. Killing of conidia was decreased after incubation of morphonuclear leukocytes with IL-4 for 2 days (P < 0.05). While superoxide anion production in response to phorbol myristate acetate was decreased by IL-4 (P < 0.05), it was not altered in response to blastoconidia and pseudohyphae. Morphonuclear leukocyte-induced pseudohyphal damage also remained unaltered. These findings suggest that IL-4 plays its detrimental role in invasive candidiasis by predominantly suppressing uptake and killing of blastoconidia by morphonuclear leukocytes. Anti-IL-4, IFN-gamma, GM-CSF and M-CSF appear to counteract suppression of morphonuclear leukocyte phagocytic activity suggesting new approaches to the management of disseminated candidiasis. PMID- 9395063 TI - Alu sequences. AB - Alu sequences are frequently encountered during study of human genomic nucleic acid and form a major component of repetitive DNA. This review describes the origin of Alu sequences and their subsequent amplification and evolution into distinct subfamilies. In recent years a number of different functional roles for Alu sequences have been described. The multiple influences of Alu sequences on RNA polymerase II-mediated gene expression and the presence of Alu sequences in RNA polymerase III-generated transcripts are discussed. PMID- 9395064 TI - SHP-1 is involved in neuronal differentiation of P19 embryonic carcinoma cells. AB - Accumulating evidence suggests that tyrosine phosphorylation plays an important role in the development of the central nervous system and in the differentiation of neuronal cells. To identify protein tyrosine phosphatases (PTPs) that might regulate signaling events leading to neuronal cell differentiation, we cloned PTP genes from the murine P19 embryonic carcinoma cell line and examined the change of their expression during differentiation. P19 cells are known to be pluripotent and the aggregate formation and subsequent replating in the presence of retinoic acid (RA) induce growth arrest and neuronal differentiation. The results demonstrated that among several PTP genes expressed in P19 cells, a cytosolic Src homology region 2 domain-containing PTP, SHP-1, is expressed highly in undifferentiated P19 cells, but is reduced to an undetectable level at day 3 after replating in the presence of RA. Further, SHP-1 was tyrosine-phosphorylated and activated at day 1 after replating. When ectopic SHP-1 was constitutively expressed, P19 cells continued to proliferate and failed to differentiate upon stimulation with RA. Collectively, these results suggest that the regulated expression and activity of SHP-1 may be involved in the neuronal differentiation of P19 cells. PMID- 9395065 TI - Overexpression of hIGF-1 exclusively in skeletal muscle increases the number of dihydropyridine receptors in adult transgenic mice. AB - The number of dihydropyridine receptors (DHPR) and sarcoplasmic reticulum (SR) Ca2+ release channels (RyR1) and their interaction determine the efficacy of the sarcolemmal excitation-SR Ca2+ release-contraction coupling (ECC). Both receptors play a central role in ECC as demonstrated in various animal species and muscle subtypes. In the present work we studied the effect of transgenic overexpression of human insulin-like growth factor 1 (hIGF-1) on the levels of these two Ca2+ channels in extensor digitorum longus (EDL) (fast-twitch), soleus (slow-twitch) and pool of fast- and slow-twitch muscles from adult C57BL/6 mice. Muscles from hIGF-1 transgenic mice showed a significant increase in IGF-1 concentration (20 30-fold) and in the number of DHPR (52% increase) whereas no significant change in RyR1 binding sites was detected. The differential effect on DHPR and RyR1 resulted in a 30% increase in DHPR/RyR1 ratio. Fast- and slow-twitch muscles showed 50 and 70% increase in the number of DHPR and 30 and 80% increase in DHPR/RyR1, respectively. These results support the concept that the increased autocrine/paracrine secretion of hIGF-1 exerts potent stimulatory effects on DHPR alpha1 subunit expression in adult skeletal muscle. PMID- 9395066 TI - Increased expression of alpha-enolase in c-jun transformed rat fibroblasts without increased activation of plasminogen. AB - Two-dimensional gel electrophoresis was used to identify polypeptides differentially expressed between normal and c-jun transformed rat fibroblasts. The level of a 49 kDa polypeptide was 3-fold elevated in c-jun transformed cells. Sequence analysis by ion trap mass spectrometry identified the polypeptide as rat alpha-enolase. Enolase functions as a cell surface receptor for plasminogen, suggesting that upregulation may increase plasminogen activation and cell surface proteolysis important for tumor growth. However, no difference was observed between normal and transformed cells in formation of plasmin, suggesting that upregulation of alpha-enolase may contribute to an increased metabolic capacity, but not to increased plasminogen activation. PMID- 9395067 TI - Functional assessment of the yeast Rvs161 and Rvs167 protein domains. AB - Mutations in RVS161 and RVS167 yeast genes induce identical phenotypes associated to actin cytoskeleton disorders. The whole Rvs161 protein is similar to the amino terminal part of Rvs167p, thus defining a RVS domain. In addition to this domain, Rvs167p contains a central glycine-proline-alanine rich domain and a SH3 domain. To assess the function of these different domains we have expressed recombinant Rvs proteins in rvs mutant strains. Phenotype analysis has shown that the RVS and SH3 domains are necessary for phenotypical complementation, whereas the GPA domain is not. Moreover, we have demonstrated that the RVS domains from Rvs161p and Rvs167p have distinct roles, and that the SH3 domain needs the specific RVS domain of Rvs167p to function. These results suggest that Rvs161p and Rvs167p play distinct roles, while acting together in a common function. PMID- 9395069 TI - Expression of Reticulomyxa filosa tubulins in Pichia pastoris: regulation of tubulin pools. AB - We expressed the alpha2- and beta2-tubulin isoforms of the giant freshwater amoeba Reticulomyxa filosa in the methylotrophic yeast Pichia pastoris. Single expression lead to little or no detectable material. Coexpression of both tubulins, however, resulted in a significant increase of expressed proteins. At the same time, the detectable internal tubulins of the host yeast cell were downregulated. This finding indicates the functionality of the expressed amoeba tubulins. Further regulation phenomena were observed on the level of equilibrium between the two R. filosa tubulin isoforms and on the level of the total tubulin pool. The P. pastoris/R. filosa system therefore seems to be an accessible system for the simultaneous study of the various mechanisms involved in tubulin regulation. PMID- 9395068 TI - A mechanism for the proarrhythmic effects of cisapride (Propulsid): high affinity blockade of the human cardiac potassium channel HERG. AB - Cisapride (Propulsid) is a gastrointestinal prokinetic agent commonly used to treat nocturnal heartburn as well as a variety of other gastrointestinal disorders. The use of cisapride has been associated with acquired long QT syndrome and ventricular arrhythmias such as torsades de pointes which produces sudden cardiac death. These cardiotoxic effects can be due to blockade of one or more types of K+ channel currents in the human heart. For this reason we compared the effects of cisapride on two cloned human cardiac K+ channels, Kv1.5 and the human ether-a-go-go-related gene (HERG) stably transfected into mammalian cells. Using patch clamp electrophysiology, we found that cisapride was a potent inhibitor of HERG displaying an IC50 value of 44.5 nmol/l when tail currents at 40 mV were measured following a 2 s test depolarization to +20 mV. When HERG currents were measured at the end of prolonged (20 s) depolarizing steps to +20 mV, the apparent affinity of cisapride was increased and measured 6.70 nmol/l. The main effect of cisapride was to enhance the rate of HERG current decay thereby reducing current at the end of the voltage clamp pulse. Furthermore, the potency of cisapride for the HERG channel was similar to that observed for the class III antiarrhythmic agent dofetilide (IC50 = 15.3 nmol/l) and the nonsedating antihistamine terfenadine (IC50 = 56.0 nmol/l). In contrast to its effects on HERG, cisapride inhibited Kv1.5 channel currents weakly displaying an IC50 value of 21.2 micromol/l. It is concluded that cisapride displays specific, high affinity block of the human cardiac K+ channel HERG. It is likely that this interaction underlies the proarrhythmic effects of the drug observed under certain clinical settings. PMID- 9395070 TI - Ro31-8220 inhibits protein kinase C to block the phorbol ester-stimulated release of choline- and ethanolamine-metabolites from C6 glioma cells: p70 S6 kinase and MAPKAP kinase-1beta do not function downstream of PKC in activating PLD. AB - The use of bisindolylmaleimide derivatives of staurosporine as selective inhibitors of protein kinase C (PKC) is in doubt following the report by Alessi [FEBS Lett. 402 (1997) 121-123] that Ro31-8220 and GF109203X are potent in vitro inhibitors of p70 S6 kinase and mitogen-activated protein kinase-activated protein kinase-1beta, as well as of PKC. Here we show that the phorbol ester stimulated release of choline- and ethanolamine-metabolites from C6 glioma cells due to phospholipid hydrolysis by phospholipase D (PLD) is not inhibited by rapamycin or PD98059, specific inhibitors respectively of p70 S6 kinase and MAPKK (MEK) and thus of MAPKAP kinase-1beta but is still completely blocked by Ro31 8220. We conclude therefore that p70S6k and MAPKAP kinase-1beta as well as MAPK are not involved in signalling pathways downstream of PKC that regulate phorbol ester-stimulated phospholipid turnover and that the inhibitory action of Ro31 8220 occurs by blocking PKC which regulates at least one pathway to PLD activation. The PI-3 kinase inhibitor, wortmannin, inhibits the phorbol ester stimulated release of ethanolamine- but not choline-metabolites from C6 cells suggesting that different PLD isoforms regulate the turnover of PtdEth and PtdCho in C6 cells. Both PLD isoforms are activated via PKC but the PtdEth-PLD is also regulated via a wortmannin-sensitive pathway. PMID- 9395071 TI - Modulation of HERG affinity for E-4031 by [K+]o and C-type inactivation. AB - Rectification of HERG is due to a rapid inactivation process that has been labeled C-type inactivation and is believed to be due to closure of the external mouth of the pore. We examined the effects of mutation of extracellular residues that remove C-type inactivation on binding of the intracellularly acting methanesulfonanilide drug E-4031. Removal of inactivation through mutation reduced drug affinity by more than an order of magnitude. Elevation of [K+]o in the wild-type channel reduces channel affinity for E-4031. Elevation of [K+]o also interferes with the extracellular pore mouth closure associated with C-type inactivation through a 'foot in the door' mechanism. We examined the possibility that [K+]o elevation reduces drug binding through inhibition of C-type inactivation by comparing drug block in the wild-type and inactivation-removed mutant channels. Elevation of [K+]o decreased affinity in both channel constructs by a roughly equal amount. These results suggest that [K+]o alters drug binding affinity independently of its effects on C-type inactivation. They further suggest that inhibition of pore mouth closure by elevated [K+]o does not have same effect on drug affinity as mutations removing C-type inactivation. PMID- 9395072 TI - Genome methylation of the marine annelid worm Chaetopterus variopedatus: methylation of a CpG in an expressed H1 histone gene. AB - Hydrolysis by methylation-dependent restriction enzymes shows that the genomic DNA of the polychaete annelid worm Chaetopterus variopedatus is methylated. Electrophoretic analyses of the digestion products indicate that the degree of methylation is lower in adult tissues than in sperm and embryonic DNA. 5 Methylcytosine was identified by HPLC, absorption spectroscopy and mass spectrometry analyses of free bases obtained by acid hydrolysis of the DNA. An average value of 1.6% methylated cytosines was determined in sperm DNA. Partial methylation was also found in an actively expressed H1 histone gene. This is the first time that genomic DNA methylation is demonstrated to occur in a worm. PMID- 9395073 TI - Expression of messenger RNA for the prostaglandin D receptor in the leptomeninges of the mouse brain. AB - The localization of prostaglandin D receptor in the mouse brain was examined by in situ hybridization histochemistry. The autoradiography showed significant hybridization signals of mRNA for prostaglandin D receptor in the leptomeninges covering the surface of the brain, but not in neurons or glia in the brain parenchyma. This finding was confirmed by Northern blot analysis using mRNA prepared from either the whole brain with the leptomeninges, brain parenchyma without the leptomeninges or the leptomeninges alone. A weak signal corresponding to the major 3.5-kbp transcript was detected in the whole brain. This band was significantly enriched in the leptomeninges, but was not detected in the brain parenchyma. These results suggest that prostaglandin D receptor is most highly, if not exclusively, expressed in the leptomeninges of the mouse brain. PMID- 9395074 TI - Regulatory phosphorylation of plant phosphoenolpyruvate carboxylase: role of a conserved basic residue upstream of the phosphorylation site. AB - In order to mimic regulatory phosphorylation of the Ser-15 of maize C4-form phosphoenolpyruvate carboxylase (PEPC), we replaced Ser-15 and Lys-12 with Asp (S15D) and Asn (K12N), respectively, by site-directed mutagenesis. Although both mutant enzymes were catalytically as active as the wild-type PEPC, they showed much less sensitivity to malate, an allosteric inhibitor, similarly to the phosphorylated wild-type PEPC. A maize protein kinase of 30 kDa which is known to be specific to PEPC (PEPC-PK), phosphorylated K12N as well as the wild-type PEPC but not S15D. The phosphorylation of K12N further diminished the sensitivity to malate. Thus, a positive charge of the conserved Lys-12 is not required for the recognition by PEPC-PK but contributes to the intrinsic sensitivity to malate inhibition. PMID- 9395075 TI - Nef protein of HIV-1 induces apoptotic cytolysis of murine lymphoid cells independently of CD95 (Fas) and its suppression by serine/threonine protein kinase inhibitors. AB - The Nef protein of HIV-1 is suggested to play a role in depletion of uninfected CD4+ T cells leading to the development of AIDS. The recombinant soluble Nef protein was shown to bind to cell surfaces of various murine lymphoid cell lines, including T and B lymphocytes, mastocytoma cells and macrophages. Cross-linking of the cell-bound Nef protein with anti-Nef antibodies induced apoptotic cytolysis of the cells. Although primary lymphocytes from young mice resisted Nef binding and Nef-induced cytolysis, treatment of the cells with concanavalin A or phytohemagglutinin made them susceptible to these activities, indicating that cellular activation is required for the apoptosis. The Nef-induced apoptosis also occurred with murine cells not expressing CD95 (Fas). These findings were quite similar to those obtained for human blood cells, suggesting that the mouse is applicable for analysis of Nef activities. The Nef-induced apoptosis was efficiently suppressed by serine/threonine protein kinase inhibitors, H7, fasudil hydrochloride and M3, which did not inhibit CD95 (Fas)-mediated apoptosis. On the other hand, bisindolylmaleimide, a protein kinase C inhibitor which inhibits CD95 (Fas)-mediated apoptosis, did not affect Nef-induced apoptosis. These results suggest that the Nef-induced apoptosis of murine cells involved a serine/threonine protein kinase-dependent signal transduction pathway distinct from the CD95 (Fas)-mediated system. PMID- 9395076 TI - The human alpha3b is a 'full-sized' laminin chain variant with a more widespread tissue expression than the truncated alpha3a. AB - We report the molecular cloning of the human laminin alpha3b chain variant and its mRNA expression pattern in adult human tissues when compared to the alpha3a variant. The mRNA encoding for the alpha3b variant is about 11 kb and the predicted translation product carries the complete set of domains typical for a 'full-sized' laminin alpha chain. Apart from the similar domain structure of alpha3b also the sequence of alpha3 resulted more closely related to the alpha5 than to the alpha4 chain. Quantitative analysis of the RNA expression in a broad panel of adult human tissues indicated that the alpha3b variant is more widely distributed than the alpha3a shorter variant. PMID- 9395077 TI - Synthesis of a new zinc finger peptide; comparison of its 'code' deduced and 'CASTing' derived binding sites. AB - Using two synthetic oligonucleotides, we have constructed a new gene containing three zinc finger motifs of the Cys2-His2 type. We named this artificial gene 'Mago'. The Mago nucleotide triplets encoding the amino acid positions, described to be crucial for DNA binding specificity, have been chosen on the basis of the proposed recognition 'code' that relates the zinc finger's primary structure to the DNA binding target. Here we demonstrate that Mago protein specifically binds the 'code' DNA target, with a dissociation constant (Kd) comparable to the Kd of the well known Zif268 protein with its binding site. Moreover, we show that the deduced Mago 'code' and the 'experimental' selected DNA binding sites are almost identical, differing only in two nucleotides at the side positions. PMID- 9395078 TI - Kinetics of the inhibition of mitochondrial respiration by NO. AB - The kinetics of the inhibition of mitochondrial respiration by NO was examined in isolated mitochondria (here obtained from rat brown adipose tissue). The Ki of NO for the inhibition was approximately 27 nM; the IC50 of NO increased in proportion to the square of an increase in O2 tension. The Km of O2 for respiration was approximately 16 microM; in the presence of NO, the dependence of respiration on O2 tension had a Hill coefficient of approximately 2. The unusual kinetics is probably related to the ability of cytochrome c oxidase to use 2 NO or 1 O2 as electron acceptor. The interaction between NO and O2 in the control of respiration could be described by the formula VO2(O2, NO) = VO2max x ([O2]2/((16 microM x (1 + [NO]/27 nM))2 + [O2]2)). Thus, the kinetics is such that respiration in the presence of physiological levels of NO is very sensitive to decreasing O2 tension. PMID- 9395079 TI - Insulin and cyclic AMP act at different levels on transcription of the L-type pyruvate kinase gene. AB - We previously demonstrated that, in hepatocytes in primary culture, the role of insulin on induction of L-type pyruvate kinase (L-PK) gene expression was mainly to induce glucokinase synthesis, needed for glucose phosphorylation to glucose 6 phosphate. However, we show here that when hepatocytes have been isolated from rats starved for 72 h, glucose and constitutive glucokinase expression was not sufficient to fully stimulate the L-PK promoter, low insulin concentrations being still required. In addition, activation remains sensitive to cAMP inhibition, but cannot be reproduced in the absence of insulin by a competitive cAMP antagonist. We propose that both insulin and cAMP act on expression of the L-PK gene at, at least, two levels: positive and negative regulation of glucokinase gene expression, and more downstream levels. PMID- 9395080 TI - The macrophage migration inhibitory factor MIF is a phenylpyruvate tautomerase. AB - A macrophage migration inhibitory factor (MIF), originally described as a product of activated lymphocytes, has been defined as a 12 kDa protein, expressed in a wide variety of tissues. Here MIF is identified as a phenylpyruvate tautomerase (EC 5.3.2.1) having p-hydroxyphenylpyruvate and phenylpyruvate as its natural substrates. The definition of MIF as an enzyme may yield insight into the mechanism of action of this proinflammatory and immunomodulating cytokine. PMID- 9395081 TI - Generation and characterization of transgenic mice expressing a human mutant alpha-galactosidase with an R301Q substitution causing a variant form of Fabry disease. AB - Transgenic mice expressing a human mutant alpha-galactosidase with an R301Q substitution, which was found in a patient with a variant form of Fabry disease, were established. The mice transcribed a sufficient amount of alpha-galactosidase mRNA, but the steady-state levels of the enzyme protein were decreased in liver, kidney and heart, only residual activity being detected in these tissues. The mice will be useful for the clarification of the defective regulation of the structurally altered enzyme protein expressed by the mutant gene at the organ or individual level as well as for the evaluation of drugs that stabilize and/or activate the mutant alpha-galactosidase. PMID- 9395082 TI - Populating the equilibrium molten globule state of apomyoglobin under conditions suitable for structural characterization by NMR. AB - Conditions have been determined under which the equilibrium molten globule state of apomyoglobin is stable and remains monomeric for periods of time sufficient for the application of three-dimensional heteronuclear NMR experiments. The quality of initial two-dimensional NMR spectra suggests that sequence-specific assignments can be made for a majority of the protein resonances under these conditions. A pH titration of the protein followed using two-dimensional 1H-15N correlation experiments indicates that the equilibrium intermediate undergoes fast exchange on the chemical shift time scale with the unfolded state and intermediate time scale exchange with the native state, and suggests a strategy to assist with backbone resonance assignments. The conditions and techniques described may be applicable to the characterization of other equilibrium folding intermediates. PMID- 9395083 TI - Thermodynamic characterizations of an intramolecularly hydrogen bonded C5 structure across proteinogenic residue. AB - Thermodynamic investigations of a smallest possible intramolecularly hydrogen bonded C5-structure, across a Thr residue, in model peptides Boc-Xxx-Thr-NH2 (Xxx = Ile, 1 or Leu, 2), indicated unusual thermal stability of the structure in non polar medium. An analysis of van't Hoff plots, constructed from variable temperature 1H NMR data, yielded the thermodynamic parameters of a hydrogen bonded five-membered ring. The non-significance of the spatial organizations of the preceding CdeltaH3 bearing hydrophobic proteinogenic residue on the thermal stability of the C5-structure has been observed. The results revealed that the contribution of this element of secondary structure is quantifiable and the stability appeared to be roughly comparable to other intramolecularly hydrogen bonded reverse turn structures frequently observed in polypeptides and proteins. PMID- 9395084 TI - Involvement of integrins and the cytoskeleton in modulation of skeletal muscle glycogen synthesis by changes in cell volume. AB - Muscle glycogen synthesis is modulated by physiologically relevant changes in cell volume. We have investigated the possible involvement of integrin extracellular matrix interactions in this process using primary cultures of rat skeletal muscle subject to hypo- or hyper-osmotic exposure with integrin binding peptide GRGDTP to disrupt integrin actions and the inactive analogue GRGESP as control. Osmotically induced increases (77%) and decreases (34%) in glycogen synthesis (D-[14C]glucose incorporation into glycogen) were prevented by GRGDTP (but not GRGESP) without affecting glucose transport. Cytoskeletal disruption with cytochalasin D or colchicine had similar effects to GRGDTP. Osmotically induced modulation of muscle glycogen synthesis involves integrin-extracellular matrix interactions and cytoskeletal elements, possibly as components of a cell volume 'sensing' mechanism. PMID- 9395085 TI - Cloning of two novel human importin-alpha subunits and analysis of the expression pattern of the importin-alpha protein family. AB - The import of many proteins into the nucleus is mediated by the importin alpha/beta heterodimer. While only one importin-beta gene has been found, several forms of importin-alpha have been described. In addition to the three human importin-alphas already identified, we report here the primary structure of two new human importin-alpha proteins. The five known human importin-alpha subunits can be classified into three subfamilies that appear conserved in higher eukaryotic organisms. We show by immunoblotting that the different importin-alpha subfamilies are expressed in a variety of human tissues and mammalian cell lines. PMID- 9395086 TI - Proteins interacting with the molecular chaperone hsp70/hsc70: physical associations and effects on refolding activity. AB - We investigated several hsp70/hsc70 interacting proteins and established by two independent techniques that hsp40 and Hop/p60 specifically interact with the 257 residue carboxy-terminal domain of hsp70 while Hap-46 and Hip/p48 bind the 383 residue amino-terminal ATP binding domain. Hap-46 and Hip/p48 competed for binding to hsc70, while Hap-46 had no effect on the binding of either Hop/p60 or hsp40 to hsc70. Hap-46 inhibited the refolding of thermally denatured firefly luciferase in an hsc70 and hsp40 dependent assay, and this effect was largely compensated by Hop/p60. These interacting proteins thus appear to cooperate in affecting the chaperoning activity of hsp70/hsc70. PMID- 9395087 TI - Identification of the yeast ACR1 gene product as a succinate-fumarate transporter essential for growth on ethanol or acetate. AB - The protein encoded by the ACR1 gene in Saccharomyces cerevisiae belongs to a family of 35 related membrane proteins that are encoded in the fungal genome. Some of them are known to transport various substrates and products across the inner membranes of mitochondria, but the functions of 28 members of the family are unknown. The yeast ACR1 gene was introduced into Escherichia coli on an expression plasmid. The protein was over-produced as inclusion bodies, which were purified and solubilised in the presence of sarkosyl. The solubilised protein was reconstituted into liposomes and shown to transport fumarate and succinate. Its physiological role in S. cerevisiae is probably to transport cytoplasmic succinate, derived from isocitrate by the action of isocitrate lyase in the cytosol, into the mitochondrial matrix in exchange for fumarate. This exchange activity and the subsequent conversion of fumarate to oxaloacetate in the cytosol would be essential for the growth of S. cerevisiae on ethanol or acetate as the sole carbon source. PMID- 9395088 TI - Photoaffinity labelling of P-glycoprotein catalytic sites. AB - Photoaffinity labelling of hamster P-glycoprotein was carried out after trapping of radioactive Mg-8-azido-ADP in the catalytic sites by vanadate or beryllium fluoride. With either trapping agent the same labelled peptide was obtained in homogeneous form, with the sequence -FNEVVFNxPTRPDI-, corresponding to residues 1034-1037 in the C-terminal nucleotide binding site. The missing residue 'x' corresponds to Tyr-1041, which is therefore a primary reaction target of 8-azido ADP. This tyrosine is conserved in all hamster, mouse and human P-glycoproteins. A second major labelled peptide fraction was also identified. The major sequence in this fraction was -NIHFSxPSR-, corresponding to residues 393-401 of hamster P glycoprotein, where 'x' corresponds to Tyr-398 in the N-terminal nucleotide binding site. Therefore Tyr-398, which is also conserved in other P glycoproteins, is also a reaction target for 8-azido-ADP. In sequence alignment of the two nucleotide binding sites, Tyr-398 exactly corresponds to Tyr-1041. The data indicate that these two tyrosines lie close to the adenine ring of bound substrate MgATP in the respective catalytic sites of P-glycoprotein. PMID- 9395089 TI - Influence of a NH2-terminal extension on the activity of KTX2, a K+ channel blocker purified from Androctonus australis scorpion venom. AB - A cDNA encoding a short polypeptide blocker of K+ channels, kaliotoxin 2 (KTX2), from the venom of the North African scorpion Androctonus australis was expressed in the periplasmic space of Escherichia coli. KTX2 was produced as a fusion protein with the maltose binding protein followed by the recognition site for factor Xa or enterokinase preceding the first amino acid residue of the toxin. The fully refolded recombinant KTX2 (rKTX2) was obtained (0.15-0.30 mg/l of culture) and was indistinguishable from the native toxin according to chemical and biological criteria. An N-extended analogue of KTX2 exhibiting three additional residues was also expressed. This analogue had 1000-fold less affinity for the 125I-kaliotoxin binding site on rat brain synaptosomes than KTX2. Conformational models of KTX2 and its mutant were designed by amino acid replacement using the structure of agitoxin 2 from Leiurus quinquestriatus as template, to try to understand the decrease in affinity for the receptor. PMID- 9395090 TI - The 5'-untranslated region of the human muscle acylphosphatase mRNA has an inhibitory effect on protein expression. AB - The cDNA of the human muscle type acylphosphatase was isolated and characterized. The mRNA presents a very long 5'-untranslated region, covering the first half of the molecule: 175 bases of this part were cloned and prediction of the possible secondary structure showed that a very stable stem-loop structure could be formed in that region. Moreover, an additional AUG triplet was found upstream of the start codon of the protein, defining an open reading frame of 60 codons which overlapped that of acylphosphatase. The possible regulatory effect on translation of this part of the mRNA molecule was studied by means of transient transfection experiments: a 10-fold decrease in the expression of a reporter protein and a dramatic decrease in the corresponding mRNA was observed, due to the presence of the 5'-untranslated region of acylphosphatase mRNA. Mutagenesis of the upstream AUG triplet eliminated mRNA instability, leading to the hypothesis that the product of the upstream open reading frame could play a role in this mechanism. PMID- 9395091 TI - Modulation of membrane activity of amphipathic, antibacterial peptides by slight modifications of the hydrophobic moment. AB - Starting from the sequences of magainin 2 analogs, peptides with slightly increased hydrophobic moment (mu) but retained other structural parameters were designed. Circular dichroism investigations revealed that all peptides adopt an alpha-helical conformation when bound to phospholipid vesicles. Analogs with increased mu were considerably more active in permeabilizing vesicles mainly composed of zwitterionic lipid. In addition, the antibacterial and hemolytic activities of these analogs were enhanced. Correlation of permeabilization and binding indicated that the activity increase is predominantly caused by an increased membrane affinity of the peptides due to strengthened hydrophobic interactions. PMID- 9395092 TI - Design of mu selective opioid dipeptide antagonists. AB - We have recently designed potent delta selective opioid antagonist dipeptides on the basis of a simple conformational analysis. Following a similar procedure we found a mu selective dipeptide antagonist, 2,6-dimethyl-Tyr-D-Phe-NH2. Although its selectivity is not as high as those of the quoted delta selective dipeptides it has good in vitro activity and looks very promising for further development since the 2,6-dimethyl-Tyr-D-Phe message, like the delta selective 2,6-dimethyl Tyr-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid counterpart, seems able to impart antagonism to longer peptides. PMID- 9395093 TI - An electrospray ionisation mass spectrometry (ESI-MS) study to probe the metal ion binding site in the DNA binding domain of the yeast transcriptional activator GAL4. AB - Electrospray ionisation mass spectrometry (ESI-MS) is used to detect metal ions and their stoichiometry of binding in the DNA binding domain of GAL4. In this analysis, the mass spectra of the apo- and metallo-proteins differ by both mass and charge, precluding the possibility of random adduct formation. Deuterium exchange NMR experiments of Zn(II)-GAL4(7-49) indicate that the binuclear metal ion structure, which is shown to have a net negative charge of -2, is the recipient of several hydrogen bonds, notably from the main-chain amide protons of the ligating cysteine residues, indicating the charge is stabilised in this manner. PMID- 9395094 TI - New biomarker evidence of oxidative DNA damage in patients with non-insulin dependent diabetes mellitus. AB - Urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG) has been reported to serve as a sensitive biomarker of oxidative DNA damage and also of oxidative stress. We have investigated oxidative DNA damage in patients with non-insulin-dependent diabetes mellitus (NIDDM) by urinary 8-OHdG assessments. We determined the total urinary excretion of 8-OHdG from 24 h urine samples of 81 NIDDM patients 9 years after the initial diagnosis and of 100 non-diabetic control subjects matched for age and gender. The total 24 h urinary excretion of 8-OHdG was markedly higher in NIDDM patients than in control subjects (68.2 +/- 39.4 microg vs. 49.6 +/- 37.7 microg, P = 0.001). High glycosylated hemoglobin was associated with a high level of urinary 8-OHdG. The increased excretion of urinary 8-OHdG is seen as indicating an increased systemic level of oxidative DNA damage in NIDDM patients. PMID- 9395095 TI - FT-IR study of the Ca2+-binding to bovine alpha-lactalbumin. Relationships between the type of coordination and characteristics of the bands due to the Asp COO- groups in the Ca2+-binding site. AB - Fourier-transform infrared spectroscopy (FT-IR) was applied to examine relationships between the type of coordination and the COO- antisymmetric and symmetric stretches of the COO- groups in the Ca2+-binding site of bovine alpha lactalbumin. The peaks at 1593, 1578, 1425, and 1403 cm(-1) were assigned to the COO- groups of Asp-82, 87, and 88 coordinating to Ca2+ in the pseudo bridging mode, according to the results of X-ray crystallography. The bands due to the COO groups were quite similar to each other between alpha-lactalbumin and EDTA which is the model compound for the pseudo bridging state. PMID- 9395097 TI - Cognitive-behavioral profiles among different categories of orofacial pain patients: diagnostic and treatment implications. AB - Psychological homogeneity in temporomandibular disorders (TMD) is not conclusive. The multidimensional pain inventory (MPI) has previously identified 3 cognitive behavioral profiles in TMD and chronic pain patients. Our aims were to replicate these findings in another cultural setting and relate the profiles to the diagnosis and to the treatment demand and outcome. The MPI was administered to 112 referrals comprising 6 categories of patients diagnosed with TMD or intractable orofacial pain. Dysfunctional profiles (high in pain and distress) were most common in patients with orofacial pain of obscure origin and more common in myofascial pain patients than in patients with other TMD diagnoses. Interpersonally-distressed profiles were found in all categories. Among patients with disk displacement, the 3rd profile (adaptive copers with low pain and distress and high control and activity) was most common in earlier successfully diskectomized patients and least common in those about to undergo invasive interventions. A dysfunctional profile was associated with treatment failure, conservative or surgical, and with the demand for radical therapy. Some support for a cyclical causality between pain and psychological factors was found. It is concluded that the robustness of the MPI as a relevant assessment instrument was further strengthened. PMID- 9395096 TI - Molecular cloning and characterization of a human brain ryanodine receptor. AB - We have cloned and sequenced the cDNA of the human brain ryanodine receptor (RyR3), which is composed of 4866 amino acids and shares characteristic structural features with the rabbit RyR3. Northern blot analysis shows that the human RyR3 mRNA is abundantly expressed in hippocampus, caudate nucleus and amygdala as well as in skeletal muscle. The human RyR3 mRNA is also detected in several cell lines derived from human brain tumors. Functional expression of RyR3 and a chimeric RyR suggests that RyR3 forms a calcium-release channel with a very low Ca2+ sensitivity. PMID- 9395098 TI - Psychological characteristics of patients applying for treatment in a dental fear clinic. AB - The aim of the present study was to assess psychological characteristics of highly anxious dental patients. Subjects were 321 patients (203 women) applying for treatment at a dental fear clinic in The Netherlands. Patients filled out several questionnaires to assess the amount of psychological complaints (Dutch version of the Symptom Checklist 90-Revised; SCL-90) and dental anxiety (Dental Anxiety Scale, Short version of the Dental Anxiety Inventory) approximately 4 months prior to their first appointment. The total mean score on the SCL-90 for the highly anxious patients was 142.7 (SD=47.2) for men and 166.5 (SD=64.3) for women. The SCL-90 total score and the mean scores of all subscales were significantly higher, and above the 80th percentile for most of the subscales, than the mean scores of the Dutch general population. The results indicate that patients who apply for treatment at a dental fear clinic are not just dentally anxious; they show complaints on a wide range of other psychological dimensions. PMID- 9395099 TI - "Checkerboard" versus culture: a comparison between two methods for identification of subgingival microbiota. AB - The present study compared the "checkerboard" DNA-DNA hybridization methodology with culture techniques for the analysis of the composition of the subgingival microbiota. 70 subjects, presenting with a variety of periodontal conditions, contributed with a total of 283 subgingival plaque samples analyzed with respect to the following species: Porphyromonas gingivalis, Prevotella intermedia/Prevotella nigrescens, Fusobacterium nucleatum, Campylobacter rectus, Eikenella corrodens, Bacteroides forsythus, Actinobacillus actinomycetemcomitans, Streptococcus sanguis and Streptococcus mutans. Species identification and quantification was performed by (i) the checkerboard method, using whole genomic, digoxigenin labeled DNA probes; and (ii) culture, including non-selective and selective media in combination with routine biochemical testing using commercial test panels. We found that the checkerboard technology resulted in higher prevalence figures for half of the species tested when compared to culture data. If the latter were used as the reference, checkerboard detection sensitivities ranged from 0.17 to 0.86, specificities from 0.17 to 1.0, and diagnostic accuracies from 0.51 to 0.81, depending on bacterial species. The use of the checkerboard data as the reference resulted in detection sensitivities for the culture procedures between 0.24 and 1.0 and specificities between 0.21 and 0.87. The checkerboard methodology resulted in statistically significant higher bacterial counts for the majority of the species. It was further observed that, for most species, the higher the total number colony-forming units in the sample, the higher the discrepancy between the results obtained by the two techniques. PMID- 9395100 TI - Expression of T-cell receptor Vbeta2, 6 and 8 gene families in chronic adult periodontal disease. AB - Substantial evidence exists to suggest a role for T-cells in periodontal disease. As yet, however, the T-cell receptors remain to be characterised at the molecular level. The expression and the nucleotide sequence of genes from the T-cell receptor beta variable (TCRBV) gene families 2, 6 and 8 were analyzed in periodontal tissue from 24 patients with chronic adult periodontal disease (CAPD) and peripheral blood lymphocytes (PBL) of 16 of these patients. A restriction in the expression of these TCRBV gene families was detected in periodontal tissue from 14/24 patients with CAPD, and the pattern of gene expression was often different between individual patients; however there was no restriction in TCRBV gene expression in matched PBL samples from 8 of these 14 patients. Quantitative RT PCR analysis of samples from 5 CAPD patients who expressed all 3 TCRBV gene families in their periodontal tissues did not reveal any significant differences in the levels of gene expression in periodontal tissue and PBL. In contrast to the findings with some CAPD patients, genes from all 3 TCRBV families were always expressed in periodontal tissue and PBL from disease-free control subjects. PCR products from both the PBL and periodontal tissue of CAPD patients were cloned and sequenced; analysis of the nucleotide sequence revealed diversity with respect to the expression of TCRB joining (TCRBJ) and TCRB diversity (TCRBD) genes and the sequence of the junctional region in all samples analysed. In conclusion, in CAPD, the pattern of TCRBV gene expression in periodontal tissue is often but not always different from that in PBL and healthy periodontal tissue, which may indicate, in some cases, a local influence on particular T-cell subsets which is relevant to the pathogenesis of periodontal disease. However, the expressed TCRB genes are heterogeneous at the nucleotide level, emphasising the underlying complexity at the molecular level in the local T-cell response in CAPD. PMID- 9395101 TI - Dentin sialoprotein (DSP) transcripts: developmentally-sustained expression in odontoblasts and transient expression in pre-ameloblasts. AB - Dentin sialoprotein (DSP), a 53 kDa glycoprotein, is believed to be present exclusively in dentin. Using rat and mouse digoxigenin labeled (DIG)-DSP and 35S DSP riboprobes, and in situ hybridization techniques, we have studied the presence of DSP mRNA at specific developmental stages of dentinogenesis. In mouse and rat molars and incisors, DSP transcripts were localized in young odontoblasts associated with early stages of predentin formation, as well as in mature odontoblasts, cells with cytoplasmic extensions embedded in the forming dentin. No DSP transcripts were detected in dental pulp, enamel organ, ameloblasts, epithelial root sheath, Meckel's cartilage, alveolar bone or tibia. Furthermore, no DSP mRNA was observed in other soft tissues including heart, lung, kidney, intestine, eye, and muscle. In addition to the intense and prolonged expression by odontoblasts, DSP mRNA was transiently expressed by pre-ameloblasts in both developing molars and incisors. These observations are consistent with the results of previous immunohistochemical studies (1). The transient expression of DSP in pre-ameloblasts across from young odontoblasts suggests an involvement of DSP in epithelial-mesenchymal interactions that are crucial to later stages of tooth development. PMID- 9395102 TI - Closing of dentinal tubules by Gluma desensitizer. AB - Gluma Dentin Bond is an adhesive system, where the primer contains 5% glutaraldehyde and 35% hydroxyethyl methacrylate. Practitioners have reported a strong desensitizing effect of the Gluma system on dentin. This study, thus, sought to evaluate the effect of this system on dentin using various microscopic techniques. 12 non-restored human molars extracted for prosthodontic reasons were used. Prior to extraction the buccal cusps were removed such that a 2 mm x 2 mm wide dentin surface was exposed. The surfaces were treated in 6 ways: (1) application of Gluma 2 cleanser, Gluma 3 primer to which 0.1% w/v fluorescein was added, and Gluma 4 sealer; (2) as in (1) but treatment with H2O/0.1% w/v fluorescein instead of the Gluma 3; (3) as in (1) but without Gluma 2; (4) as in (1) but with application of 5% glutaraldehyde instead of Gluma 3; (5) as in (1) but without Gluma 4; (6) as in (1) but with application of 35% HEMA/0.1% w/v fluorescein instead of Gluma 3. Following extraction, 1 tooth per procedure was prepared for confocal laser scanning microscopy. The remaining teeth were fixed and prepared for SEM and TEM evaluation. In specimens of procedures (1) and (5), tubular occlusions could be seen to a depth of 200 microm. In specimens of procedure (4) tubular occlusions were found only to a depth of 50 microm. Such occlusions were not seen in control specimens (2), in specimens where the smear layer had not been removed (3), or following application of HEMA alone (6). It is concluded that glutaraldehyde can intrinsically block dentinal tubules. The septa in the tubules may counteract the hydrodynamic mechanism for dentinal sensitivity. PMID- 9395103 TI - Transmission electron microscopic study of in vivo pellicle formation on dental restorative materials. AB - This electron microscopic investigation was performed to examine the ultrastructure of the in vivo formed salivary pellicle on 15 different dental materials. Test pieces of amalgam alloys, casting alloys, titanium, ceramics, resins, composite resins, glass polyalkenoate cement, and bovine enamel were attached to the buccal and lingual surfaces of the upper first molars in three adults using removable intraoral splints. The splints were carried over periods of 2 and 6 h. Pellicle-like structures could be identified on all tested material surfaces by transmission electron microscopic analysis. The pellicle layer showed a high degree of similarity on different kinds of surfaces with regard to the ultrastructural appearance. However, distinct differences could be detected in the ultrastructural pattern and thickness of the pellicle layer formed on buccally and lingually mounted specimens. Pellicles adsorbed on buccally carried test pieces were characterized by a heterogeneous, globular appearance and a thickness ranging from 500 to 1,000 nm after 6 h. In contrast, all lingually mounted test pieces were covered by a granular pellicle of about 100 nm thickness after 6 h. It is concluded that the ultrastructural pattern and extent of salivary pellicle formation are influenced by locally available salivary biopolymers and locally effective shearing forces rather than by material dependent parameters. PMID- 9395104 TI - Improved efficacy of dentin-bonding agents. AB - Dentin cavities, prepared in extracted human teeth, were treated with various proprietary dentin-bonding agents and then filled with a light-cured restorative resin for posterior use. All bonding agents were either treated in accordance with the manufacturers' instructions or combined with Gluma, which is an aqueous solution of glutaraldehyde and HEMA, a hydrophilic monomer. 10 min after polymerization, the width and the extent of the marginal contraction gap was measured approximately 0.1 mm below the free surface of the filling, using a light microscope. With nearly all dentin-bonding agents, the marginal contraction gap could be significantly reduced if Gluma was used after conditioning of the dentin. The reason for this improvement may be that glutaraldehyde cross-links the collagen fibers and thereby strengthens the organic part of the hybrid layer, however, other mechanisms might also play a role in the improvement found. PMID- 9395105 TI - Chondroitin sulfate isomers in synovial fluid of healthy and diseased human temporomandibular joints. AB - Synovial fluid was collected from the superior articular cavity of the temporomandibular joint in patients with unilateral internal derangement and joint pain whose contralateral joint was healthy. Glycosaminoglycans were liberated by digestion with pronase E, and precipitated with cetylpyridinium chloride and ethanol. Unsaturated disaccharide isomers of chondroitin sulfate, obtained following chondroitinase ACII digestion, were analyzed by high performance liquid chromatography. Analytic data indicated that deltaDi-0S and deltaDi-6S were often found in chondroitin sulfate from the fluid of the diseased joints. The amounts of deltaDi-0S and deltaDi-6S differed significantly between synovial fluid samples from the diseased and healthy joints. Comparison of the relative proportions of the unsaturated disaccharides in the synovial fluid with previously reported values for several tissues, indicated that the chondroitin sulfate originated from articular cartilage, with possibly some contributions from soft connective tissues and serum present in the synovial fluid. These results suggest that chondroitin sulfate in the synovial fluid provides a useful indicator of the degree of internal derangement of the temporomandibular joint. PMID- 9395106 TI - Irradiation effects on microhardness of fluoridated and non-fluoridated bovine dentin. AB - The objective of this study was to evaluate the effects of irradiation on microhardness of dentin. Dentin blocks from the cervical region of bovine incisors were treated as follows: (1) no irradiation; (2) irradiation of specimens up to 60 Gy (2 Gy/day, 5 days/week); (3) no irradiation, but fluoridation of specimens for 5 min/d; (4) irradiation of specimens and daily fluoridation. Knoop hardness number (KHN) of the control specimens was 62.63+/ 14.75 (mean+/-SD). This was significantly different from the irradiated dentin samples (8.74+/-2.59 KHN). Hardness of the fluoridated dentin specimens was 11.19+/-1.95 KHN in the non-irradiated group and 10.03+/-2.76 KHN in the irradiated groups, respectively. Within the limitations of an in vitro study, it is concluded that dentin is severely affected by irradiation. This could be an explanation for the frequently observed side-effects of irradiation like loss of enamel, gap formation at the amelodentinal junction, and caries of the cervical region. Fluoridation with acidic gels decreases microhardness of dentin surface, and does not prevent softening due to radiation, when saliva is absent. PMID- 9395107 TI - Research revisited: Gunnar Rolla's contribution to oral health. AB - Gunnar Rolla's contributions to oral science cover an enormous range. They include research into the basic understanding of pellicle and plaque formation, to mechanisms for the prevention of oral diseases. Rolla has collaborated with over 100 persons from many different countries and thus his influence on dental research has been global. PMID- 9395108 TI - On the value of research in health care. AB - This paper describes 3 important myths that reduce the political impact of academic research in healthcare and reviews data on the social return of such research. The myths are (i) scientists' interest in pursuing issues without concern for value to society; (ii) industry and private investors should fund more of society's basic research; (iii) Academic research can be improved by better administration. A recent study on return on investment in academic research in general is discussed as well as more specific studies on research related to healthcare. Examples include the BCG and Polio vaccines that today saves Norway more in healthcare costs than the entire public bill for drugs. Some factors that explain why research is undervalued are reviewed, notably rapid changes making benchmarking difficult, insufficient routines in accounting, changing goals, and unpredictable outcomes. PMID- 9395109 TI - The end of science? PMID- 9395110 TI - Review on fluoride, with special emphasis on calcium fluoride mechanisms in caries prevention. AB - Low concentrations of fluoride have a beneficial effect on enamel and dentin de- and remineralization. After fluoride treatments, such as topical applications, rinses or dentifrices, salivary fluoride concentrations decrease exponentially in a biphasic manner to very low concentrations within a few hours. For treatments to be effective over periods longer than the brushing and the following salivary clearance, fluoride needs to be deposited and slowly released. Calcium fluoride (or like) deposits act in such a way, owing to a surface covering of phosphate and/or proteins, which makes the CaF2 less soluble under in vivo conditions than in a pure form in inorganic solutions. Moreover, due to the phosphate groups on the surface of the calcium fluoride globules, fluoride is assumed to be released with decreasing pH when the phosphate groups are protonated in the dental plaque. PMID- 9395111 TI - Fluoride-induced precipitates on enamel surface and subsurface areas visualised by electron microscopy and confocal laser scanning microscopy. AB - The present study examined the enamel surface after in vitro topical treatments with a neutral 2% NaF solution. For minimising the risk of artefacts, samples were inspected without pre-treatment as fresh, naturally wet specimens by complementary techniques: variable pressure electron microscopy (VP-SEM) and confocal laser scanning microscopy (CLSM). VP-SEM provided information on the surface morphology, whereas CLSM allowed non-destructive visualisation of subsurface areas. Neutral NaF solutions induced globular precipitates on the enamel surfaces. If the globules formed may be described as "calcium fluoride like material", the additional information of this experiment is that, after interaction with neutral solutions, they also contain considerable amounts of NaF. When the NaF solutions were soaked on pellets and then were brought into contact with the enamel surfaces, NaF crystallites of cubic shape are formed. Confocal optical tomographies of subsurface enamel after treatments with neutral NaF solutions revealed partly coroded enamel structures, whereas VP-SEM showed intact surfaces. In between the coroded areas, a fine granulate precipitate could be observed. This is evidence that fluoride induces the formation of sub-surface precipitates only when applied during demineralisation. The precipitate could be readily removed by 24-h contact with a KOH solution. PMID- 9395112 TI - The resistance of titanium tetrafluoride-treated human enamel to strong hydrochloric acid. AB - The acid resistance of TiF4-treated enamel was investigated to establish a possible treatment modality for endogenous dental erosion. Enamel slabs were prepared from human molars and treated with solutions of TiF4. Scanning electron microscopy (SEM) and microhardness testing were used to examine the effects of the exposure of the treated enamel to strong acid. SEM micrographs showed the presence of heavy deposits on enamel surfaces. The surface coating, formed following TiF4 application, appeared to be resistant to severe acid attacks. Microhardness measurements showed that TiF4 treatment inhibited enamel softening. It is concluded that topical TiF4 application may be effective in prevention of dental erosion caused by hydrochloric acid from the stomach in patients with frequent vomiting or gastroesophageal reflux. PMID- 9395113 TI - Fluoride in plaque fluid and saliva after NaF or MFP rinses. AB - A micro-analytic method, capable of measuring the fluoride concentration in 5 nl of plaque fluid, was used to follow changes in fluoride concentration in saliva and plaque fluid at 6 single tooth-sites in 6 subjects for 180 min after a 0.048 M fluoride rinse as a NaF or MFP (sodium monofluorophosphate) solution. The maximum fluoride concentrations in saliva after NaF was 13x higher than with MFP. About 5% of the total amount of fluoride following the 20 ml NaF rinse was retained in the oral cavity. The corresponding figure followig MFP was <1%. The saliva/plaque fluid fluoride ratios for upper molars and lower incisors were significantly higher than for the upper incisors and lower molars. There was a tendency for a decline in the ratios with respect to time for all sites. To characterize the plaque fluid fluoride intra-oral single-site distribution and clearance, fluoride concentration versus time (AUC) was calculated from 10 to 60 min after a rinse. The NaF AUC followed the order: upper incisor, lower molar, upper molar and lower incisors reflecting a different exposure and clearance pattern due to the different access of the plaque to saliva. The MFP AUC values varied more, but were all significantly lower than the NaF AUC values. Analysis of plaque fluid fluoride curves at various sites revealed an exponential decline in most cases. With NaF, the baseline plaque fluid fluoride levels were not reached within 3 h. It is concluded that NaF solutions result in a significantly higher intra-oral fluoride exposure than MFP solutions. The fluoride distribution and clearance of fluoride from different sites in the oral cavity are linked to salivary access to these sites. These site-specific differences may have clinical consequences with regard to the dynamics of fluoride in the de- and remineralization processes. PMID- 9395114 TI - On the role of human salivary micelle-like globules in bacterial agglutination. AB - The hypothesis to be tested in this in vitro study was that the salivary micelle like globules (SMGs) have a role in the agglutination of some oral bacteria. An attempt to determine the mechanisms for the interactions involved was also carried out. 4 laboratory and 4 native streptococci strains were tested. Human whole (HWS) and parotid (HPS) saliva was collected from 4 subjects, and SMGs were isolated from both salivas, and agglutination was recorded in the various bacterial suspensions over time. HPS, HWS and SMGs isolated from HPS and HWS caused typical agglutination patterns for the mutans strains. Salivary supernatants (without SMGs) caused a much delayed or no agglutination. Electron microscopy showed SMG-like structures on the surface of the agglutinated bacteria. Addition of pyrophosphate to HPS prevented agglutination, whereas guanidine HCl prevented normal agglutination of a sanguis strain, and urea had no obvious effect. Together, these results indicate that the SMGs are important in the agglutination of streptococci, and that both calcium-dependent, electrostatic and hydrophobic interactions may be involved. PMID- 9395115 TI - Fractionation of salivary micelle-like structures by gel chromatography. AB - Globular structures have been demonstrated in human parotid saliva by transmission electron microscopy and photon correlation spectroscopy. The aim of this study was to fractionate these salivary globular structures for analytical and preparative purposes using a gel-filtration material capable of separating spherical particles up to 300-400 nm in diameter. Freshly obtained parotid saliva was applied to a Sephacryl S-1000 column. Peak fractions were collected and prepared for transmission electron microscopy (TEM) or for amino acid analysis. Bovine milk was included as the casein micelles by TEM appear to be similar to the salivary aggregates and their elution profiles are known. The salivary globular structures were eluted in one major peak. TEM of negatively stained samples from the peak fractions demonstrated globular protein aggregates consistent with the salivary structures in parotid saliva. Amino acid analysis showed characteristic amino acid profiles with unusual high levels of proline, 40 45%. The casein micelles were eluted in one major peak and separated from the whey proteins. This study indicates that the salivary globular structures can be isolated by gel chromatography. The amino acid analysis indicates that proline rich proteins may be an important fraction of the salivary globular structures. PMID- 9395116 TI - Chemical agents for the control of plaque and plaque microflora: an overview. AB - This presentation provides an overview of the technologies available for the chemical control of plaque. It is generally accepted that the formation of dental plaque at the interfaces of tooth/gingiva is one of the major causes of gingival inflammation and dental caries. Several therapeutic approaches have been used to control dental plaque and supragingival infections. These include fluoride preparations such as stannous fluoride, oxygenating agents, anti-attachment agents, and cationic and non-cationic antibacterial agents. Among the fluoride preparations, stable stannous fluoride pastes and gels have been shown to reduce supragingival plaque, gingivitis, hypersensitivity and caries. The effect of the oxygenating agents on the supragingival plaque has been equivocal, but recent data indicate that a stable agent which provides sustained active oxygen release is effective in controlling plaque. A polymer, PVPA, which reduced attachment of bacteria to teeth was shown to significantly reduce plaque formation in humans. A new generation of antibacterials includes non-ionics such as triclosan, which in combination with a special polymer delivery system, has been shown to reduce plaque, gingivitis, supragingival calculus and dental caries in long-term studies conducted around the world. Unlike the first generation of agents, the triclosan/copolymer/sodium fluoride system is effective in long-term clinicals and does not cause staining of teeth, increase in calculus, or disturbance in the oral microbial ecology. PMID- 9395117 TI - Dental calculus: recent insights into occurrence, formation, prevention, removal and oral health effects of supragingival and subgingival deposits. AB - Dental calculus, both supra- and subgingival occurs in the majority of adults worldwide. Dental calculus is calcified dental plaque, composed primarily of calcium phosphate mineral salts deposited between and within remnants of formerly viable microorganisms. A viable dental plaque covers mineralized calculus deposits. Levels of calculus and location of formation are population specific and are affected by oral hygiene habits, access to professional care, diet, age, ethnic origin, time since last dental cleaning, systemic disease and the use of prescription medications. In populations that practice regular oral hygiene and with access to regular professional care, supragingival dental calculus formation is restricted to tooth surfaces adjacent to the salivary ducts. Levels of supragingival calculus in these populations is minor and the calculus has little if any impact on oral-health. Subgingival calculus formation in these populations occurs coincident with periodontal disease (although the calculus itself appears to have little impact on attachment loss), the latter being correlated with dental plaque. In populations that do not practice regular hygiene and that do not have access to professional care, supragingival calculus occurs throughout the dentition and the extent of calculus formation can be extreme. In these populations, supragingival calculus is associated with the promotion of gingival recession. Subgingival calculus, in "low hygiene" populations, is extensive and is directly correlated with enhanced periodontal attachment loss. Despite extensive research, a complete understanding of the etiologic significance of subgingival calculus to periodontal disease remains elusive, due to inability to clearly differentiate effects of calculus versus "plaque on calculus". As a result, we are not entirely sure whether subgingival calculus is the cause or result of periodontal inflammation. Research suggests that subgingival calculus, at a minimum, may expand the radius of plaque induced periodontal injury. Removal of subgingival plaque and calculus remains the cornerstone of periodontal therapy. Calculus formation is the result of petrification of dental plaque biofilm, with mineral ions provided by bathing saliva or crevicular fluids. Supragingival calculus formation can be controlled by chemical mineralization inhibitors, applied in toothpastes or mouthrinses. These agents act to delay plaque calcification, keeping deposits in an amorphous non-hardened state to facilitate removal with regular hygiene. Clinical efficacy for these agents is typically assessed as the reduction in tartar area coverage on the teeth between dental cleaning. Research shows that topically applied mineralization inhibitors can also influence adhesion and hardness of calculus deposits on the tooth surface, facilitating removal. Future research in calculus may include the development of improved supragingival tartar control formulations, the development of treatments for the prevention of subgingival calculus formation, the development of improved methods for root detoxification and debridement and the development and application of sensitive diagnostic methods to assess subgingival debridement efficacy. PMID- 9395118 TI - Oral mucosal side effects of cytotoxic chemotherapy of testicular cancer. A retrospective study. AB - Cancer chemotherapy often leads to injury of normal cells. Adverse effects on oral mucosa have been documented for several cytotoxic treatment regimens. The aim of the present retrospective study was to evaluate incidence and degree of oral soft tissue side-effects of a cisplatin-based chemotherapy regimen used for treating testicular cancer. The study was based upon a questionnaire mailed to 56 consecutive patients treated at the Norwegian Radium Hospital. A total of 39 individuals joined the study, as 2 patients refused and 15 did not reply. The patients were divided into two groups, a case group (24 individuals) having received 4 7 cycles of cisplatin-based chemotherapy in addition to surgery, and a control group (15 individuals) treated with surgery alone. The study revealed that 62% of the patients in the chemotherapy group developed adverse soft tissue reactions, with mucositis and pain as chief complaints, whereas none in the control group experienced any mucosal complications. PMID- 9395119 TI - The influence of triclosan, zinc or propylene glycol on oral mucosa exposed to sodium lauryl sulphate. AB - Previous studies on triclosan treatment of skin exposed to sodium lauryl sulphate (SLS) indicated a protective role of zinc and an irritant effect of propylene glycol (PG). The aim was hence to examine whether zinc or PG also may affect SLS induced oral mucosal reactions, and also to test the influence of zinc in combination with triclosan. 15 healthy dental students participated in this double-blind crossover study performed in 2 experimental series. They were rinsing 2x daily with solutions containing (A) 1.5% SLS, (B) 1.5% SLS/0.5% zinc citrate and (C) 1.5% SLS/PG (1:8) in experiment 1, and (D) 1.5% SLS/0.15% triclosan/0.3% zinc citrate and (E) 1.5% SLS/0.15% triclosan in experiment 2. Clinical evaluation by 2 examiners of degree of erythema and oral mucosal desquamations was then performed. The critical micellar concentration was also determined. SLS and SLS/PG, which were not different in effect, evoked significantly more erythematous reactions than SLS/Tri/Zn. This solution was numerically but not statistically better than SLS/Tri, and the latter also did lead to significantly less erythema than SLS/PG. In conclusion, the present study revealed no irritation of the oral mucosa due to PG, whereas a protective effect of zinc as well as the anti-inflammatory effect of triclosan were confirmed. PMID- 9395120 TI - On the transformation of sulfur-containing amino acids and peptides to volatile sulfur compounds (VSC) in the human mouth. AB - Halitosis is most often caused by oral conditions. Volatile sulfur compounds (VSC), constituting the major components of oral malodor, are produced by anaerobic, gram-negative bacteria retained mainly in periodontal pockets or on the tongue dorsum. Sulfur-containing amino acids serve as substrate for these bacteria. VSC have also been found to have unfavorable effect on the tissue. The aim of this study was to examine whether normal, healthy individuals with no history of halitosis were able to produce VSC from cysteine, when applied as a mouthrinse. A further aim of the study was to investigate and compare the potential of other sulfur-containing amino acids and peptides as substrates for oral VSC production and to localize the odor-production sites. A portable sulfide monitor was used for VSC registration. Results showed that all test subjects produced high oral concentrations of VSC upon rinses with cysteine, which thus seems to be a major substrate for VSC production. The other sulfur-containing substrates had much less effect. It was found that the tongue was the major site for VSC production, and that saliva per se caused low VSC production. PMID- 9395121 TI - Neurological effects of microwave exposure related to mobile communication. AB - Due to the wide and growing use of mobile communication, there is increasing concern about the interactions of electromagnetic radiation with the human organism, and, in particular, with the brain. In the present report, experimental studies on putative electrophysiological, biochemical and morphological effects of continuous or pulsed microwave radiation are briefly reviewed. Such effects have been described in vitro and in vivo using animals and humans. Particularly, effects on neuronal electrical activity, cellular calcium homeostasis, energy metabolism, genomic responses, neurotransmitter balance and blood-brain barrier permeability have been reported. However, some results have either been disputed, since experimental replication led to contradictory findings, or been related to procedural side effects. Since neurological disturbances induced by mobile telephone devices would be of considerable interest for public health, the authors recognize that further experimental studies, involving strict positive and negative control conditions, will be required in the future. At the present state of knowledge there is no positive evidence that pulsed or continuous microwave exposure in the non-thermal range confers elevated risk to the health of the brain. PMID- 9395122 TI - Risk factors for microangiopathy-related cerebral damage in the Austrian stroke prevention study. AB - Microangiopathy-related cerebral damage (MARCD) represents a common incidental MRI observation in the elderly. The risk factors of such findings are widely unknown. We therefore performed MRI in 349 randomly selected volunteers (ages 50 to 70 years) without neuropsychiatric disease, and evaluated the association of MARCD with conventional and recently suggested cerebrovascular risk factors such as apolipoprotein E genotypes, plasma concentrations of essential antioxidants and anticardiolipin antibody titres. MARCD was defined as evidence of early confluent and confluent deep white matter hyperintensities and lacunes. It was present in 71 (20.3%) subjects. Individuals with MARCD were older than those without such findings (62.7 years vs 59.6 years; P=0.0001). They had a higher rate of arterial hypertension (45.1% vs 28.1%; P=0.006) and cardiac disease (50.7% vs 37.1%; P=0.04), higher systolic blood pressure readings at exam (144.4 mmHg vs 136.7 mmHg; P=0.004), and higher serum fibrinogen concentrations (327.1 mg/dl vs 292.5 mg/dl; P=0.001). Their levels of total cholesterol (217.6 mg/dl vs 231.2; P=0.009), apolipoprotein A-I (167.3 mg/dl vs 177.4 mg/dl, P=0.02), lycopene (0.17 micromol/l vs 0.24 micromol/l; P=0.003), retinol (1.91 micromol/l vs 2.10 micromol/l; P=0.02) and alpha-tocopherol (27.55 micromol/l vs 31.14 micromol/l; P=0.001) were significantly lower. Forward stepwise regression analysis created a model of independent predictors of MARCD with age entering first (odds ratio 2.01/10 years), fibrinogen second (odds ratio 2.45/100 mg/dl), alpha-tocopherol third (odds ratio 0.55/10 micromol/l), and arterial hypertension fourth (odds ratio 1.96). The association of MARCD with various treatable clinical conditions may have preventive implications. PMID- 9395123 TI - Serial brain CT in corticobasal degeneration: radiological and pathological correlation of two autopsy cases. AB - This report concerns serial brain computed tomography (CT) images in two patients with corticobasal degeneration (CBD). The diagnosis of CBD was confirmed by neuropathological examination. In one case, we obtained serial brain CT images for one year and five months, and in the other, for three years and four months. The CT images showed that the anterior portions of the cerebrum atrophied progressively with the length of the disease. This was also seen in the CT images with respect to the caudate nucleus. Regarding radiological and pathological correlations, we consider that the progressive atrophy of the anterior portions of the cerebrum, seen in brain CT images, is ascribed to neuronal loss and gliosis in the anterior portions of the cerebrum. We also believe that the observed caudate nucleus atrophy is due to loss of neurons and gliosis in this structure, and also to the fibrillary gliosis of the cerebral white matter. The progressive atrophy of the caudate nucleus detected in brain CT is a valuable feature for the neuroradiological diagnosis of CBD. PMID- 9395124 TI - Tolerance induction to myelin basic protein by intravenous synthetic peptides containing epitope P85 VVHFFKNIVTP96 in chronic progressive multiple sclerosis. AB - Peptide-based tolerance induction may be useful for antigen-specific immunotherapy of human autoimmune diseases. Induction of tolerance to myelin basic protein (MBP) was examined in a Phase I clinical trial in multiple sclerosis (MS) patients with chronic progressive disease using a peptide that is immunodominant for MBP specific T cells and B cells. Tolerance induction was monitored by quantification of MBP specific autoantibodies in cerebrospinal fluid (CSF). The route of peptide administration was important since only intravenous but not intrathecal or subcutaneous injection induced tolerance to MBP. Following a single intravenous injection of a peptide containing epitope P85VVHFFKNIVTP96, MBP autoantibodies were undetectable for three to four months. Tolerance was more prolonged following a second injection since autoantibodies were low or undetectable after one year in the majority of patients. Duration of tolerance to MBP depended on MHC class II haplotypes of patients; tolerance was long-lived in all patients with disease associated HLA-DR2. No neurological or systemic side effects were observed, regardless of the route of peptide administration. These data demonstrate that intravenous administration of a soluble peptide can result in long-lasting tolerance to an autoantigen in humans. PMID- 9395125 TI - Normal human aging: factors contributing to cerebral atrophy. AB - Factors that accelerate rates of 'normal' age-related cerebral atrophic and degenerative changes are important because they may predispose to cognitive declines. To determine characteristic patterns of normal aging, risk factors were correlated with serial neurological-neuropsychological examinations, CT measures of progressive cerebral atrophy, local tissue hypodensities, or perfusional declines. Both cross-sectional and longitudinal designs were utilized. Ninety four cognitively and neurologically normal aging volunteers, 15 with a history of transient ischemic attacks (TIAs), were followed for mean intervals of 3.0+/-2.1 years. Results indicated that: (1) after age 60, cerebral atrophy, polio- and leuko-araiosis doubled and cerebral perfusion decreased, with marked individual variations; (2) risk factors independently accelerating cerebral atrophy and cortico-subcortical perfusional declines included TIAs, hypertension, smoking, hyperlipidemia, excessive alcohol consumption and male gender; (3) progressive leuko-araiosis correlated directly with cortical atrophy and cortical perfusional declines. We posit that: (1) cerebral atrophy and degenerative changes result from neuronal shrinkage and/or loss, which are accelerated by TIAs, hypertension, smoking, hyperlipidemia, excessive alcohol consumption and male gender; (2) accelerated cerebral atrophic and degenerative changes identified by neuroimaging should be considered as markers for depleted neuronal synaptic reserves, which predispose to cognitive declines. Interventions available for controlling some of these risk factors include control of TIAs, hypertension, and hyperlipidemia, as well as tobacco and alcohol withdrawal. PMID- 9395126 TI - Tumor necrosis factor alpha and its receptors in relapsing-remitting multiple sclerosis. AB - In the attempt to further characterize the extent and timing of tumor necrosis factor (TNF)alpha-system activation during multiple sclerosis (MS), we performed a cross-sectional and a longitudinal study in a total of 73 relapsing-remitting MS patients. We assessed serum levels of soluble TNFalpha, soluble TNFalpha receptor 1 (R1) and soluble TNFalpha receptor 2 (R2) in 65 relapsing-remitting MS patients in different phases of disease. TNFalpha, R1 and R2 serum levels measured in MS patients did not differ from those measured in healthy individuals and did not correlate with (a) clinical relapses, (b) presence of gadolinium enhancing brain-magnetic resonance imaging (MRI) lesions, and (c) bioactivity of TNFalpha. We also measured in 8 additional relapsing-remitting MS patients peripheral blood mononuclear cells (PBMC) mRNA levels of TNFalpha, R1 and R2 every 15 days for one year. In 4 of these patients we also measured levels of soluble TNFalpha, R1 and R2 every 15 days for 5 months across a clinical exacerbation. PBMC TNFalpha, R1 and R2 mRNA levels and serum levels of soluble R1 and R2, but not TNFalpha, fluctuated concordantly (P<0.05) and peaked a mean of 6 weeks before clinical and MRI evidence of disease activity. Moreover, we found a significant positive correlation between cumulative TNFalpha and R2 mRNA levels (measured during the follow-up period in the 8 MS patients studied serially) and the number of clinical attacks recorded in these patients during the study. Our data show that serum levels of soluble TNFalpha, R1, and R2 in MS patients do not differ from those of healthy individuals. However, although within normal values, the transcription and production rate of all these molecules fluctuate concordantly in the peripheral blood during the course of the disease (with the exception of soluble TNFalpha) and their maximal elevation significantly precedes the occurrence of clinical exacerbations. It is not clear whether soluble TNFalpha escapes recognition by commonly used assays or is simply not released in its soluble form in MS patients. In any case, measurement of TNFalpha mRNA levels and R1 and R2 mRNA and protein levels appears to be a better indicator of disease fluctuations during the course of MS than assessments of soluble TNFalpha protein. PMID- 9395128 TI - Correlates of p53- and Fas (CD95)-mediated apoptosis in Alzheimer's disease. AB - Apoptosis may be an important mechanism of cell loss in Alzheimer's disease (AD). Experimentally, apoptosis is preceded by nuclear accumulation of p53, and increased expression of Fas (CD95) antigen. In the present study, quantitative Western blot analysis of postmortem frontal and temporal lobe tissue demonstrated significantly higher mean levels of p53 and Fas in AD relative to age-matched controls. Immunohistochemical staining and in situ apoptosis assays demonstrated increased p53 and Fas expression and DNA fragmentation in overlapping populations of cortical neurons, and cortical and white matter glial cells distributed in regions damaged by neurodegeneration. Double-label immunohistochemical staining studies revealed p53 immunoreactivity in: 1) cortical neurons without tau immunoreactive neurofibrillary tangles; 2) numerous, but not all tau immunoreactive neuropil neurites and white matter axons; 3) dystrophic fibrils surrounding amyloid-beta-immunoreactive plaques; and 4) glial cells characterized as A2B5+ protoplasmic astrocytes or oligodendrocytes. The prominent distribution of dystrophic p53-immunoreactive processes around amyloid-beta-containing plaques suggests that amyloid deposits are associated with local neuritic degeneration. In addition, the results suggest that many tau-immunoreactive neuritic processes originate from degenerating (p53) as well as regenerating neurons. Finally, apoptosis of glial cells (A2B5+) required to maintain the functional integrity of axons and dendrites may represent an important pathogenic mechanism of axonal loss and synaptic disconnection in AD. PMID- 9395127 TI - Impaired quantitative cerebral blood flow in scleroderma patients. AB - Systemic Sclerosis (SSc) is a multisystem disease characterised by proliferation of vascular tissue, obliterative microvascular lesions and diffuse organ fibrosis. Despite widespread vascular disease, Central Nervous System complaints are only infrequently reported and it is uncertain whether they merely derive from systemic complications or whether they may be also caused by a primary vascular process within the brain. Regional cerebral blood flow (rCBF) was quantitatively measured by the 133Xenon clearance technique in twenty-seven consecutive SSc patients without relevant systemic complications and with different severity of vascular involvement, as staged by nailfold capillary videomicroscopy (NCV). Absolute, percent, and asymmetry rCBF values were compared (z-statistics) with age- and sex-matched healthy controls. Cerebral MRI and Mini Mental State Examination (MMSE) were also performed. Doppler sonography of neck vessels and Transcranial Doppler sonography (TCD) were performed in patients presenting rCBF reduction. Cerebral hypoperfusion was found in the 52% of patients, i.e.: in 33% of patients with the 'early' NCV pattern, in 56% of patients with the 'active' pattern, and in 67% of patients with the 'late' NCV pattern. Thirty percent were the MRIs showing focal and/or diffuse signal abnormalities in the white matter of both hemispheres with the highest rate (44%) in the 'late' NCV pattern. MMSE disclosed mild dementia in one patient in the 'late' NCV group and some mistakes in 6 more patients, in the 'active' or 'late' NCV groups, whereas TCD failed to find significant stenosis of Willis' arteries. Cerebral hypoperfusion is shown for the first time in a substantial part of SSc patients without either neurological symptoms or relevant systemic complications. It is suggested that the rCBF reduction might be related to the systemic scleroderma microangiopathy although, probably due to the paucity of connective tissue in cerebral vessels, the vast majority of patients remains in a subclinical phase. PMID- 9395129 TI - Normal auditory brainstem and cochlear function in extreme pediatric plumbism. AB - Lead (Pb) intoxication in children has been associated with encephalopathy, sensory and cognitive impairments. We investigated the prevalence and neuro sensory effects of Pb exposure in children living in Andean villages of Ecuador with high Pb contamination from discarded automobile batteries used in the local ceramics glazing industry. Venous blood samples were collected from 107 children in the Pb glazing area and from 39 children living in a geographically distant area with no known Pb contamination and measured for blood lead (PbB) levels. Auditory brainstem responses (ABR) and audiological/otological tests were conducted on children in the Pb-Glazing Group. The median PbB level for children in the Pb-Glazing Group was 40.0 microg per dl (range: 6.2-128.2 microg per dl) and for the non Pb-Glazing Group 6.0 microg per dl (1.9-18.0 microg per dl). The differences in PbB levels for children in the study and control areas were statistically significant (t-test, P<0.0001). ABR tests on the Pb-Glazing Group indicated normal wave latencies and neural transmission times, and no statistical correlation between PbB level and interpeak latencies. Audiological tests showed normal cochlear function and no statistical relation between auditory thresholds and PbB level. Contrary to prevailing assumptions, elevated PbB levels in children do not invariably impair auditory brainstem neural transmission or sensory-neural cochlear function, both of which have been implicated as significant contributors to the neurodevelopmental disabilities associated with childhood plumbism. PMID- 9395130 TI - The presence of autoantibodies to N-terminus domain of GluR1 subunit of AMPA receptor in the blood serum of patients with epilepsy. AB - We have analyzed the serum from patients with refractory epilepsy for the presence of autoreactive antibodies to AMPA glutamate receptor subunits. The presence and the level of autoantibodies were assessed using immunoblot and ELISA with synthetic peptides specific for subregions of AMPA glutamate receptor subunits. Patients with refractory epilepsy exhibited strong immunoreactivity to GluR1 subunit compared to healthy donors and patients with Parkinson's disease and Alzheimer's disease. Weak immunoreactivity to other AMPA glutamate receptor subunits was also detected and the signal was diminished in the raw GluR4>GluR3>GluR2. The occurrence of autoantibodies to specific neurotransmitter subunits in the sera of patients with refractory epilepsy suggest that autoimmune process may underlie this disorder. PMID- 9395131 TI - GM1 gangliosidosis type 3 with severe jaw-closing impairment. AB - The patient had adult GM1 gangliosidosis (type 3) with severe impairment of mastication caused by dystonia of anterior digastric muscles (jaw-opener) on clenching. This is the first report on jaw dystonia severe enough to cause the masticatory impairment in adult GM1 gangliosidosis. The discordance of closing and opening muscles during mastication might be caused by a basal ganglia lesion in this disease. PMID- 9395132 TI - Successful treatment of painful traumatic mononeuropathy with carbamazepine: insights into a possible molecular pain mechanism. AB - The delayed onset of painful paresthesias following trauma to a peripheral nerve is a well recognized but poorly understood phenomenon. This report describes an illustrative case of painful paresthesias in the territory of the ilioinguinal nerve, 3 to 6 weeks after an otherwise routine herniorraphy, which subsequently responded dramatically to carbamazepine. The case is considered in light of recent studies which have determined molecular changes which occur in dorsal root ganglion (DRG) neurons following axotomy and neuroma formation. Voltage-dependent sodium (Na+) channels in DRG neurons undergo a change following axotomy, in which there is significant up- and down-regulation of different subpopulations of Na channels over a time frame measured in days to weeks. Such changes may render the DRG neurons hyperexcitable, thus contributing to a neuropathic pain syndrome, yet susceptible to treatment with a sodium channel blocker such as carbamazepine. PMID- 9395133 TI - Confirmation that neither cyanide intoxication nor mutations commonly associated with Leber's Hereditary Optic Neuropathy are implicated in Tanzanian Epidemic Optic Neuropathy. PMID- 9395134 TI - Amyotrophic lateral sclerosis and multifocal motor neuropathy with conduction block. PMID- 9395135 TI - Detection of tick-borne encephalitis virus by sample transfer, plaque assay and strand-specific reverse transcriptase polymerase chain reaction: what do we detect? AB - Experimental inoculation of mice provides a well characterized model for studying infection with tick-borne encephalitis virus (TBEV), a flavivirus pathogenic for humans. Conflicting data on the kinetics of viremia and the development of virus titers in the brain, however, were only recently shown to have resulted from the use of assay systems with different levels of sensitivity in the titration of TBEV, i.e. plaque assay or sample transfer into naive recipient mice. Theoretically, RT-PCR could extend further the detectability to antibody neutralized virus and when undertaken strand-specifically discriminate active replication from the mere presence of TBEV. We have compared the conventional methods for detection of TBEV with a newly devised RT-PCR method. As expected, RT PCR, in contrast to the infectivity assays, detected antibody-neutralized virus. Furthermore, the mere presence or active replication of the virus could be differentiated by strand-specific RT-PCR. Plaque assay and sample transfer, in contrast, both detected only infectious virus. However, whereas sample transfer provides higher sensitivity for detection of TBEV from solid organs, the plaque assay is less costly and considering animals welfare more convenient. Thus, the newly devised method may allow the resolution of unanswered questions, while both the traditional infectivity assays retain their benefits in certain situations. PMID- 9395137 TI - Use of site-directed mutagenesis to generate a herpes simplex virus type 1 strain 17+ mutant lacking seven HindIII restriction endonuclease cleavage sites. AB - The genome of herpes simplex virus type 1 (HSV-1) strain 17+ contains ten HindIII and four XbaI restriction endonuclease (RE) cleavage sites. We have previously reported the isolation of an HSV-1 mutant, 1702, devoid of all the four XbaI sites. Here we report the isolation of HSV-1 mutants lacking seven of the HindIII sites plus the four XbaI sites. In order to destroy the various HindIII sites, mutagenic oligonucleotides were synthesized and introduced in to the plasmids containing HSV-1 restriction endonuclease fragments spanning these HindIII sites. All the seven HindIII sites were removed by site-directed mutagenesis. Two methods of site-directed mutagenesis were used: 1) the HindIII site at 0.91 map coordinates (mc) of HSV-1 strain 17+ genome was deleted using a gapped, heteroduplex molecule of DNA, and 2) uracil-rich single-stranded DNA templates were used in in vitro mutagenesis reactions to remove the HindIII sites at 0.08, 0.1, two at 0.18, 0.26 and 0.64 mc. These HindIII site deletions were then marker transferred back in to the 1702 genome to generate virus mutants devoid of specific HindIII sites. No other deletions and/or insertions were observed within the viral genomes of mutant viruses as allowed by restriction endonuclease analysis of their 32P-labelled DNAs. All the HindIII site-deletion mutants, 1721 1733, showed comparable growth properties and polypeptide profiles to those of the parental 17+ and 1702 viruses. PMID- 9395136 TI - Comparison of ELISA and RT-PCR for the detection of beet yellows closterovirus in plants and aphids. AB - A reverse-transcription polymerase chain reaction (RT-PCR) was developed to detect beet yellows closterovirus (BYV) in plants and single aphids using primers spanning the conserved regions of the published sequences of the coat protein gene and open reading frames 7 and 8. Three total RNA extraction procedures were examined and all were found to produce RNA of sufficient quality for RT-PCR, although the RNA extraction kit supplied by Flowgen was found to be the most versatile for the extraction of BYV from individual aphids. When 60 aphids, which had fed on virus infected sugar beet were tested by RT-PCR, 55% of individuals were found to contain BYV. Two groups of 36 individual aphids were tested by TAS ELISA using a specific BYV monoclonal antibody; 53% gave positive absorbance values when a substrate amplification system was used, but none was found to contain virus when the system was replaced with the substrate p-nitrophenyl phosphate. An immunocapture RT-PCR method was shown to detect BYV regardless of whether the RNA had been extracted from the trapped particles or the reverse transcription and PCR mixtures were added to the wells containing intact particles. The RT-PCR method is now used to determine the numbers of BYV carrying aphids migrating into sugar-beet crops. PMID- 9395138 TI - Use of Doehlert matrices for study of poliovirus-1 adsorption. AB - Experiments designed according to Doehlert matrices were carried out to study poliovirus-1 adsorption to Na-montmorillonite in a complex aqueous environment. Salt concentration and valence, virus load, clay concentration, and organic matter concentration were included in the design as selected parameters for possible or known involvement in viral adsorption in environmental waters. Use of this experimental design not only allowed to detect and quantify direct influence of the tested parameters upon the viral response, but also to reveal the influence of interactions between these tested factors. Thus, beyond the reassessment of the higher efficiency of multivalent cations on virus adsorption, as opposed to monovalent ones, detection was enabled of a tannic acid/aluminium specific interaction that seemed to be responsible for the nonavailability of these elements for interaction with viruses. Such a statistical tool allows for a gain in experimental accuracy beyond technical improvements and is particularly suited for low-cost study of multifactorial phenomena. PMID- 9395139 TI - Immunoassays to study prevalence of antibody against GB virus C in blood donors. AB - Immunoassays were developed to determine the seroprevalence of antibody against human GB virus C (GBV-C). The antigenic target in each assay was a 44.6-kDa glycosylated protein representing the first 315 amino acids encoded by the GBV-C E2 gene. Sera or plasma were assayed for E2 antibody using an anti-human EIA format in which antigen-coated polystyrene beads were reacted with sample, and bound antibody was detected by addition of enzyme labelled goat anti-human IgG. The presence of anti-E2 antibody was confirmed using a sandwich EIA format in which samples were reacted with antigen coated polystyrene beads, followed by addition of solution phase biotinylated antigen. Detection of antibody captured biotinylated E2 was accomplished by addition of enzyme-conjugated anti-biotin antibody. Antibody against the E2 antigen was detected in 7.4 and 7.8% of 500 sera and 500 plasma, respectively, from US volunteers donating to a Wisconsin blood center, and in approximately 10.7% of hepatitis and retrovirus marker negative volunteer blood donors from a Missouri blood center. The rate in 1018 sera from US commercial donors at multiple US blood centers was 36.7%. These results indicated a relatively high prevalence of GBV-C exposure in US volunteer donors, and particularly in commercial donors. The clinical implication of the high exposure rate is unclear. These immunoassays are being combined with nucleic acid detection to assess prevalence of GBV-C world wide and to determine if GBV-C plays a role as an etiologic agent. PMID- 9395141 TI - Development of a RT-PCR test coupled with a microplate colorimetric assay for the detection of a swine Arterivirus (PRRSV) in boar semen. AB - Transmission of porcine reproductive and respiratory syndrome virus (PRRSV) through boar semen has been demonstrated, stressing the need for a reliable semen PRRSV detection test. A diagnostic assay was developed based on amplification of the PRRSV RNA by reverse transcription and polymerase chain reaction (RT-PCR) followed by detection of the amplification products by hybridization and colorimetric assay in microwell plates. A highly reproducible and efficient method of viral RNA isolation from semen samples was set up. A combined RT-PCR procedure was performed, incorporating the use of uracil-N-glycosylase (UNG) in combination with dUTP instead of dTTP to prevent false positive results due to carry-over contamination. An RNA internal control was added during the RNA isolation procedure to detect false negative results. The colorimetric detection in microwell plates of amplification products from either PRRSV or IC RNA gave specific and objective results and was automated. A cut-off value of 1000 RNA copies or 10 TCID50 of PRRSV per ml of semen samples could be detected with this assay. Semen samples collected from experimentally-infected boars were tested with this assay and showed PRRSV excretion early after infection and for an extended period. PMID- 9395140 TI - Generation of a p10-based baculovirus expression vector in yeast with infectivity for insect larvae and insect cells. AB - A new, versatile baculovirus vector was developed for the generation of recombinants in the yeast Saccharomyces cerevisiae and for the expression of foreign proteins in both insect larvae and in insect cells. This vector is based on Autographa californica multiple nucleocapsid nucleopolyhedrovirus (AcMNPV) and exploits the 10-kDa protein promoter (p10) for the expression of the foreign gene. The p10 locus was used for the insertion of a yeast-selectable marker system (ARS-URA-URA3) and of a gene for screening and titration of recombinants in insect cells (beta-galactosidase). The polyhedron-positive phenotype of this vector is maintained allowing its use in insect larvae, by feeding polyhedra, and in insect cells, by infecting with budded virus. The generation of this baculovirus vector requires a single recombination step in yeast prior to infection of insect cells, but has the advantage over the vector designed previously (Patel et al., A new method for the isolation of recombinant baculovirus, Nucleic Acids Research 20 (1992) 97-104) that these vectors can also be used in insects. PMID- 9395142 TI - Single-antibody in situ enzyme immunoassay for infectivity titration of hepatitis A virus. AB - Hepatitis A virus (HAV) establishes a persistent infection in cultured cells, with minimal effect on host cell metabolism. As a result, the virus produces very little, if any, cytopathic effect (CPE), even with cell culture-adapted strains. This feature precludes the use of a plaque or standard endpoint assay (using CPE as an indicator of infection) for the titration of infectious virus. The radioimmunofocus assay (RIFA) is the standard method for HAV titration, though this method is labour intensive and requires the use of radioisotopes. To this end, a single-antibody in situ enzyme immunoassay (EIA) has been developed, using binding of a perioxidase-labelled monoclonal antibody to fixed cell monolayers as an indicator of infection. This novel assay is highly reproducible, can be read by eye, and is suitable for high throughput situations. Furthermore, the assay has been validated against the RIFA making it suitable for use in studies validating the safety of therapeutic biologicals for human use. PMID- 9395143 TI - Disinfection of cell-associated and extracellular HIV-1 by PUVA treatment. AB - To inactivate cell-associated and extracellular HIV-1 while preserving cellular surface antigens, a procedure was used based on PUVA treatment, i.e. addition of psoralen to cell suspensions followed by irradiation with UVA light. T-lymphoid MT-4 cells were infected with HIV-1 strain NL4-3, 4'-aminomethyl-4,5',8 trimethylpsoralen was added, and the cell suspension was irradiated with 20 mW/cm2 UVA light for 3, 4 and 5 min. To evaluate virus inactivation, cells and supernatants were diluted serially and cocultured with uninfected MT-4 cells. Infectious HIV-1 was detected by cytopathic effects, immunofluorescence and p24 antigen ELISA. UVA irradiation at 3.6 J/cm2 (3 min 20 mW/cm2) reduced the amounts of both cell-associated and extracellular infectious HIV-1 by more than five orders of magnitude. Even at more stringent conditions of PUVA treatment (10 min 20 mW/cm2 UVA irradiation), conformational cellular surface epitopes remained detectable by flow cytometry. PMID- 9395145 TI - Detection of antibodies against iridoviruses in the serum of the amphibian Bufo marinus. AB - Sera from the amphibian Bufo marinus (cane or marine toad) were investigated using a newly-developed enzyme-linked immunosorbent assay for the detection of antibodies to ranaviruses (Family Iridoviridae). Epizootic haematopoietic necrosis virus (EHNV) or Bohle iridovirus (BIV) was affinity purified from cell culture supernatants and simultaneously bound to the solid phase using specific linker antibodies. After binding to antigen, antibodies in Bufo marinus serum were then detected with a specific anti-immunoglobulin reagent. Of 21 Bufo marinus sera, 3 contained antibodies against EHNV, BIV or related viruses in the ranavirus group, at titres greater than 3200. Reactivity of cane toad antibodies against BIV, an amphibian virus, was greater than that against similar concentrations of EHNV, a piscine virus. PMID- 9395144 TI - Rapid direct diagnosis of mumps meningitis by ELISA capture technique. AB - ELISA capture technique (ELISAc) was carried out using a rabbit hyperimmune serum attached to a solid phase for capturing mumps antigens in cerebrospinal fluid (CSF) in patients with meningitis and/or in supernatants of infected Vero cells. A biotin-labelled rabbit serum prepared from the previous serum was added and the reaction was read by an enzymatic (avidine-peroxidase) reaction by automated reading. The cut-off was calculated in 100 CSFs negative for viruses by conventional diagnosis. The specificity was evaluated in Vero cells infected with 22 CSFs collected from vaccinated children (URABE AM9 attenuated vaccine) who developed meningitis. A guinea pig hyperimmune serum confirmed the specificity. Results in culture correlated with the ELISA capture technique (ELISAc). No cross reactivity was observed with parainfluenza 1, 2, 3 human reference strains. At least 2.5 ngs of purified mumps proteins were detected corresponding to 10(1.5) infectious particles per ml. ELISAc applied directly to 14 CSFs collected from unvaccinated children with meningitis diagnosed five positive cases, whereas in four cases conventional diagnosis had to be undertaken twice. ELISAc permitted the diagnosis of one additional patient. The test can be carried out in 3 h. PMID- 9395146 TI - The development of the somatoform dissociation questionnaire (SDQ-5) as a screening instrument for dissociative disorders. AB - Using cases of dissociative disorder (n=50) and other DSM-IV diagnoses (n=50), a somatoform dissociation self-report questionnaire was developed and its capacity to function as a screening device for dissociative disorders was analysed. A list of 75 items was constructed which, according to clinical experience and expert judgement, could reflect instances of somatoform dissociation. Statistical analyses revealed the 20 best discriminating items. Stepwise forward logistic analysis detected five items which, as a group, provided optimal discrimination between the two groups. At an estimated prevalence rate of dissociative disorders of 10% among psychiatric patients the sensitivity would be 94%, the specificity would be 96%, the positive predictive value would be 72%, and the negative predictive value would be 99%. Cross-validation in an independent sample (n=33/42) largely corroborated the initial findings. The SDQ-5 can be used as a brief screening device for dissociative disorders. PMID- 9395147 TI - First-trimester maternal gestational infection and cycloid psychosis. AB - Using a structured interview, the mothers of patients with cycloid psychosis, manic depression and controls (40 mothers in each case) were investigated in order to assess the occurrence of maternal gestational infection and other obstetric complications during pregnancy with the affected child. The cycloid psychoses with low heritability and a good long-term prognosis were found to be significantly associated with first-trimester respiratory infection (i.e. influenza and febrile cold). Furthermore, maternal infection seems to predict an early onset in cycloids. In manic depression, we failed to identify a significant link with maternal gestational infection or other obstetric complications. These findings are discussed in the light of our previous reports of an excess of maternal gestational infections during the second trimester in chronic schizophrenics. Our results suggest that the exogenously induced disturbances of fetal brain maturation during the first trimester of gestation caused by maternal respiratory infection via live virus or disturbed maternal immune response are involved in the aetiology of cycloid psychoses. PMID- 9395149 TI - Reasons for substance use in schizophrenia. AB - Abuse of and dependence on drugs, alcohol and other substances in schizophrenia are being increasingly recognized and well documented in the literature. It has been suggested that up to 60% of patients with schizophrenia use illicit drugs. A total of 41 subjects who fulfilled DSM-III-R criteria for schizophrenia and substance abuse or dependence were asked to describe their reasons for using such substances, the reasons why they might stop and the subjective effects of the substances. Drugs were reportedly used to increase pleasure, to 'get high' and to reduce depression. However, subjective effects of increased depression and positive symptoms were also reported. These results are considered in the context of potential treatment strategies. PMID- 9395148 TI - The healthy control subject in psychiatric research: impulsiveness and volunteer bias. AB - Exciting and demanding biomedical experiments may attract a specific subgroup of people as volunteers. In the present study of selection bias, subjects volunteering in a psychobiological study that included a potentially painful procedure (lumbar puncture) were compared with those who declined to participate, with regard to scores on personality scales administered during a previous investigation of the same subjects. Significant differences were found on the Eysenck Personality Questionnaire and Karolinska Scales of Personality Impulsiveness scale, suggesting an over-representation of impulsive individuals among the volunteers. If the specific subject of investigation has implications for the type of individual who will participate as a healthy volunteer in biomedical research, variation will be introduced, affecting the independent variable, and the conclusions that can be drawn from such research may be questionable. PMID- 9395150 TI - Electroconvulsive therapy vs. paroxetine in treatment-resistant depression -- a randomized study. AB - Failure to respond to adequate pharmacological treatment for major depression is now the most common indication for the use of electroconvulsive therapy (ECT). The advantages of ECT with respect to both speed and quality of response are clinically important issues, but surprisingly few studies have examined the efficacy of ECT in relation to newer antidepressant agents such as selective serotonin reuptake inhibitors (SSRIs). A total of 39 subjects with major depression and with at least two failed antidepressant trials (mean 4.9 trials) were randomized to either paroxetine treatment (n=18) or right unilateral (RUL) ECT (n=21). Up to the end of the study treatment we found a reduction in the HAMD score of 59% for the ECT group and of 29% for the paroxetine group (P<0.001 paired t-test). In the ECT group, 71% of subjects fulfilled the response criteria (at least a 50% decrease in total HAMD score). The present study found ECT to be superior to paroxetine in medication-resistant major depression, in terms of both degree and speed of response. PMID- 9395151 TI - Citalopram in the treatment of obsessive-compulsive disorder: an open pilot study. AB - Obsessive-compulsive disorder (OCD) is a common anxiety disorder, which often causes significant impairment of the affected individual's social, occupational or interpersonal functioning. Previous reports suggest that the disorder may be treated with the tricyclic antidepressant clomipramine, and also with the more recently introduced selective serotonin reuptake inhibitors (SSRIs), such as fluoxetine, fluvoxamine, sertraline and paroxetine. The present 24-week open pilot study was designed to examine the efficacy, appropriate dose range, side effects and clinical usefulness of citalopram in OCD. A total of 29 OCD patients were included in the study, of whom 76% showed alleviation of symptoms as evaluated by various self- and observer-rated scales, such as the Yale-Brown Obsessive Compulsive Scale. In most cases the citalopram doses used were in most cases 40 or 60 mg daily, and the treatment was well tolerated. The most commonly experienced adverse events during the study were nausea, vomiting, increased dreaming and decreased sleep. Diminished sexual desire and orgasmic dysfunction were also reported. Despite having the limitations of an open study, our results suggest that citalopram may be effective in the treatment of obsessive-compulsive disorder. PMID- 9395152 TI - Long-term outcome of depot neuroleptic maintenance treatment among chronic psychotic patients. AB - A total of 51 chronic psychotic out-patients, with a median age of 51 years and median duration of psychosis of 23 years, treated with depot neuroleptics, entered a 5-year follow-up study with assessments of symptoms, side-effects and plasma concentration of the depot drug (follow-up (FU) patients). The outcome for 38 non-eligible (NE) patients was obtained from hospital case reports. The relapse rate was higher for NE than for FU patients (71% vs. 50%). The mortality rate was 9%, and the median age at death was 47 years. Half of the FU patients completed 3 years of treatment uneventfully. Of the total of 89 patients, only 18% remained stable over a period of 5 years. The depot dose was approximately the same after 3 years (median 255 mg, range 50-1018 mg chlorpromazine equivalents). PMID- 9395154 TI - Suicide attempts in a cohort of drug abusers: a 5-year follow-up study. AB - A group of 125 drug abusers admitted consecutively for detoxification and short term rehabilitation were followed up 5 years after discharge. They were asked about possible suicide attempts in a semi-structured face-to-face interview. Nearly half of the group (45%) reported having attempted suicide at some point in their life. The most common reasons given were the loss of a person whom they loved, and feelings of loneliness. Only three respondents reported using their drug of choice in the attempt(s). The suicide attempters were more often found to have been in child psychiatric treatment earlier, and to have experienced loss of significant others in childhood, than those who did not report attempting suicide. At follow-up the suicide attempters indicated that they experienced more depressive moods and more severe psychological problems than those who had never made a suicide attempt. The importance of assessing the risk of suicide attempts among drug addicts in order to be able to take measures to prevent future suicidal behaviour is emphasized. PMID- 9395153 TI - Prediction of outcome and early vs. late improvement in OCD patients treated with cognitive behaviour therapy and pharmacotherapy. AB - In this study, follow-up results of cognitive-behaviour therapy and of a combination of cognitive-behaviour therapy with a serotonergic antidepressant were determined. The study also examined factors that can predict this treatment effect, both in the long term and in the short term. In addition, it investigated whether differential prediction is possible for cognitive-behaviour therapy vs. a combination of cognitive-behaviour therapy with a serotonergic antidepressant. A total of 99 patients were included in the study. Treatment lasted 16 weeks, and a naturalistic follow-up measurement was made 6 months later. Of the 70 patients who completed the treatment, follow-up information was available for 61 subjects. Significant time effects were found on all outcome measures at both post treatment measurement and follow-up. No differences in efficacy were found between the treatment conditions. Effectiveness at post-treatment measurement appears to predict success at follow-up. However, 17 of the 45 non-responders at the post-treatment measurement had become responders by the follow-up. The severity of symptoms, motivation for treatment and the dimensional score on the PDQ-R for cluster A personality disorder appear to predict treatment outcome. No predictors were found that related specifically to cognitive-behaviour therapy or combined treatment. These results indicate that the effectiveness of cognitive behaviour therapy or a combination of cognitive-behaviour therapy and fluvoxamine at the post-treatment measurement is maintained at follow-up. However, non response at post-treatment does not always imply non-response at follow-up. Patients with more severe symptoms need a longer period of therapy to become responders. Although predictors for treatment success were found, no evidence was found to determine the choice of one of the treatment modalities. PMID- 9395155 TI - Validation of the Bech-Rafaelsen Mania Scale using latent structure analysis. AB - The essential criteria of internal validity have not been sufficiently evaluated for any mania rating scale, although the fulfillment of such criteria is a prerequisite for summing the item scores to give a total score reflecting the severity of mania, and for comparing total scores across patient groups that differ with regard to variables such as age and sex. This study investigated the internal validity of the Bech-Rafaelsen Mania Scale (MAS), based on the ratings of 100 consecutively admitted drug-free DSM-III-R manic patients. Application of logistic latent structure models did not statistically confirm the additivity of the MAS. However, a modified MAS (the MAS-M) arising from the analyses fulfilled the measurement model. Transferability of the MAS-M across age and sex was also confirmed. The MAS-M showed an acceptable concurrent validity and an adequate sensitivity in discriminating between responders and non-responders among patients participating in a drug trial. The MAS-M presented here is the first mania rating scale that has been shown to fulfil statistical criteria for internal validity. PMID- 9395156 TI - Life events in childhood, adolescence and adulthood and the relationship to panic disorder. AB - The aim of this study was to explore the association between stressful life events (SLE) and the development of panic disorder (PD) in an Israeli sample. A total of 44 PD patients and a matched control group were studied with regard to SLE over the life cycle (in childhood, adolescence, adulthood and the year preceding the outbreak of the disorder). The major findings were as follows. (i) With regard to the total number of life events experienced in childhood and adolescence, the PD group had experienced significantly more life events than the control group. (ii) No differences were detected in the total amount of SLE between the PD group and the control group with regard to adulthood and the year preceding the outbreak of the disorder, although the PD group had more life events relating to loss in adulthood, whereas in the year before the outbreak of PD life events relating to 'love and family', negative and loss events were more prevalent. These results expand previous findings by demonstrating that SLE in childhood and adolescence may contribute to the development of PD in adulthood. PMID- 9395157 TI - Seizures and myoclonus associated with antidepressant treatment: assessment of potential risk factors, including CYP2D6 and CYP2C19 polymorphisms, and treatment with CYP2D6 inhibitors. AB - All adverse drug reaction reports labelled seizures or myoclonus during treatment with antidepressants and stored in the Swedish national database for spontaneous reporting of adverse drug reactions were reviewed in order to evaluate possible risk factors. The reporting physicians were contacted and asked for complementary information, and blood samples for determination of the CYP2D6 and CYP2C19 genotypes were obtained from patients available. In total, 25 cases of seizures and 7 cases of myoclonus were studied. The drugs included were maprotiline (n=8), mianserin (n=7), fluvoxamine (n=6), amitriptyline (n=3), clomipramine (n=3), citalopram (n=2), paroxetine (n=2) and lofepramine (n=1). Previously suggested predisposing factors were identified in all but four cases (87%). None of the 11 patients genotyped were found to be poor metabolizers with respect to the enzymes CYP2D6 or CYP2C19. Thus, neither the CYP2D6 nor the CYP2C19 genotype were found to be associated with the occurrence of seizures/myoclonus during treatment with antidepressants. However, 15 patients (47%) were concomitantly treated with drugs with potential inhibitory effects on CYP2D6, such as neuroleptics and dextropropoxyphene, and the patients might thus have been converted from the extensive metabolizer to the poor metabolizer phenotype during this treatment. Concomitant treatment with drugs decreasing the seizure threshold and/or inhibiting the metabolism of antidepressants appeared to be an important risk factor for the occurrence of seizures/myoclonus. PMID- 9395158 TI - Cortisol in light treatment of seasonal and non-seasonal depression: relationship between melatonin and cortisol. AB - The effect of bright light on cortisol and the relationship between melatonin and cortisol were studied in 63 depressed patients (42 patients with a seasonal pattern and 21 patients with a non-seasonal pattern). The patients were matched for age, time of treatment and severity of depression. Before and after light treatment the severity of the depression was rated with the Comprehensive Psychopathological Rating Scale (23 items) and the Hamilton Depression Rating scale (18 items), and serum cortisol and melatonin were drawn at nine time-points between 20.00 and 08.00 hours. Two hours of light treatment (350 cd m-2) was given daily for 10 days either in the morning (06.00-08.00 hours) or in the evening (18.00-20.00 hours). As reported earlier, patients with a seasonal pattern improved significantly more than patients with a non-seasonal pattern of depression, and no significant differences were found between the treatment efficacy of morning compared to evening light. A cosinor analysis showed that the cortisol batyphase was significantly advanced by morning light, but was not delayed by evening light. A delay in batyphase cortisol showed a weak significant correlation with a decrease in the absolute and relative sum of scores. The batyphase of cortisol occurred approximately 3 h earlier than the acrophase of melatonin. Of the changes in the melatonin acrophase 43% were reflected in a change of cortisol batyphase, indicating a hierarchical relationship with melatonin as the co-ordinating hormone transducing part of the information of the external light to the phase position of cortisol. No significant differences between patients with a seasonal or a non-seasonal pattern were seen in mesor, amplitude or batyphase of cortisol before treatment, and no significant changes in mesor or amplitude were seen as a result of light treatment. PMID- 9395159 TI - A comprehensive method of assessing routine CT scans in schizophrenia. AB - Morphological brain abnormalities are common in schizophrenia, although the aetiological and clinical significance of these findings is largely unknown. Substantial between-subject variability suggests that large samples are needed to study the full implications of brain pathomorphology. Computerized tomography (CT) is frequently used routinely in schizophrenia, and large numbers of scans are available for study. This article describes the development and statistical properties of a rapid and simple method of assessing CT scans. The CT Rating Scale for Schizophrenia (CTRSS) is minimally affected by variability in scanning procedures, is reliable, and accurately estimates area and volumetric measures of brain spaces. By promoting the comprehensive assessment of large numbers of routinely obtained scans, the CTRSS would allow the investigation of variables that may systematically affect results (e.g. gender and age) and variables with low prevalence. The CTRSS provides a useful adjunct to technologically more sophisticated methods of assessment such as magnetic resonance imaging (MRI). PMID- 9395160 TI - Effects of child-rearing by schizophrenic mothers: a 25-year follow-up. AB - We conducted a 25-year follow-up study of 50 children of schizophrenic mothers, consisting of 25 children reared by their mothers and 25 children reared apart. The children's adult psychiatric status was evaluated in a 3-h structured interview employing a battery of syndrome check-lists and scales. A slightly higher incidence of psychopathology (including schizophrenia-spectrum disorders) was found among the reared-apart subjects. This may possibly be attributed to their greater genetic predisposition, as suggested by their mothers' more severe illnesses. Lifetime diagnoses do not provide evidence that psychopathology in offspring at genetic risk is increased by rearing by a schizophrenic mother. PMID- 9395161 TI - The impact of treatment resistance on depressive relapse following electroconvulsive therapy. PMID- 9395162 TI - Reproducibility of ventricular fibrillation characteristics in patients undergoing implantable cardioverter defibrillator implantation. AB - INTRODUCTION: The purpose of this study was to evaluate the immediate reproducibility of local electrogram characteristics recorded during repeated episodes of induced ventricular fibrillation (VF) in patients undergoing implantable cardioverter defibrillator (ICD) implantation. METHODS AND RESULTS: Power spectral analysis (using a fast Fourier transform algorithm) of electrograms recorded during 3 seconds of VF were analyzed in 24 patients undergoing ICD implantation using a Medtronic Transvene lead. Patients had 2 to 7 episodes of VF that were induced during defibrillation threshold testing. VF was induced by burst pacing (n = 20) or T wave shock (n = 4). Simultaneous electrograms during VF were recorded from a Medtronic Transvene lead with the following configurations: (1) a narrow spaced (12 mm) dedicated bipole used clinically for sensing; (2) a unipolar electrogram from the right ventricular coil; and (3) a widely spaced (18.3 mm) integrated bipole using the distal tip and the coil. Intraclass correlation coefficients (ICCs) were determined to examine the reproducibility of these VF characteristics among VF episodes in each patient. Recordings from both bipolar configurations had ICCs from 0.40 to 0.55, whereas unipolar recordings ICCs were below 0.40. Reproducibility was similar for dedicated and integrated recordings. CONCLUSIONS: Frequency characteristics of repeated episodes of VF induced in the same subjects show fair-to-good but not excellent reproducibility. Bipolar recordings were far more reproducible than unipolar recordings, but both bipolar configurations had similar reproducibility. These findings have implications for both the pathophysiology of induced VF and the design of VF detection algorithms. PMID- 9395163 TI - Carotid sinus hypersensitivity in patients undergoing coronary arteriography: relation with the severity of carotid atherosclerosis and the extent of coronary artery disease. AB - INTRODUCTION: The purpose of the present investigation was to study the precise relationship between carotid sinus hypersensitivity (CSH) and both the severity of carotid atherosclerosis and the extent of coronary artery disease in patients who were referred for evaluation for suspected ischemic heart disease. METHODS AND RESULTS: Duplex echocardiography and coronary angiography were used to assess carotid and coronary artery atherosclerosis in 130 consecutive patients. Carotid sinus stimulation was performed before coronary arteriography with simultaneous recordings of the ECG and aortic pressure. Coronary artery disease was present in 103 patients (79%). Thirty patients (23.08%) had one-vessel disease (1-VD), 31 (23.85%) had 2-VD, 29 (22.31%) had 3-VD, and 13 patients (10%) had left main coronary artery disease. Carotid artery atherosclerosis was present in 100 patients (76.92%) and carotid disease (diameter stenosis > 50%) was present in 24 patients (18.46%). CSH was found in 33 patients (25%). The incidence of CSH was 9% in patients with carotid stenosis 1%-15%, 17% in patients with stenosis 16% 49%, 85% in patients with stenosis 50%-79%, and 100% in patients with stenosis > or = 80%. The incidence of CSH was 11%, 17%, 23%, 34%, and 62% in patients with no VD, 1-VD, 2-VD, 3-VD, and left main coronary artery disease, respectively. Stepwise multiple logistic regression analysis revealed that carotid disease and left main coronary artery disease were the most significant determinants of CSH (P < 0.001 and P = 0.013, respectively). CONCLUSION: The incidence of CSH increased in proportion to the severity of carotid and coronary atherosclerosis. These data provide evidence that CSH is closely related to severe carotid atherosclerosis or left main coronary artery disease in patients being evaluated for suspected ischemic heart disease. PMID- 9395164 TI - Comparison of the effects of regional ischemia and hyperkalemia on the membrane action potentials of the in situ pig heart. Experimental Cardiology Group, University of North Carolina at Chapel Hill. AB - INTRODUCTION: This study was designed to determine the role of increased extracellular potassium [K+]e on action potential duration (APD) in the in situ porcine heart during acute regional no-flow ischemia. METHODS AND RESULTS: In open chest, anesthetized swine, an arterial shunt from the carotid artery to the mid-left anterior descending coronary artery was created through which a solution of KCl was infused to raise [K+]e. Myocardial [K+]e was determined by potassium sensitive electrodes, and transmembrane action potential was recorded by floating glass microelectrode. During the first 2 minutes of ischemia, APD at 90% repolarization (APD90) lengthened by 31.2 +/- 1.1 msec (P < 0.05). The comparable increase in [K+]e alone shortened APD90. During the next 6 minutes of ischemia, [K+]e rose to 11.3 +/- 0.3 mM and APD90 showed a decrease. However, the comparable increase in [K+]e by infusion of KCl caused further shortening of APD90 at similar levels of [K+]e. CONCLUSIONS: Acutely ischemic myocardium showed a brief increase in APD90 during the first 2 minutes of ischemia, followed by a fall in APD90 after 2 minutes of ischemia, but the shortening is less than anticipated by the rise in [K+]e. Thus, we hypothesize that other component(s) of ischemia may inhibit action potential repolarization. PMID- 9395165 TI - Extracellular potassium and the action potential duration of the ischemic heart. PMID- 9395166 TI - Differential effects of D-sotalol, quinidine, and amiodarone on dispersion of ventricular repolarization in the isolated rabbit heart. AB - INTRODUCTION: Increased dispersion of ventricular repolarization has been suggested as a cause of proarrhythmic effects of Class IA or III antiarrhythmic drugs, such as d-sotalol, quinidine, and amiodarone. METHODS AND RESULTS: The influence of d-sotalol, quinidine, and amiodarone on the dispersion of monophasic action potential (MAP) durations was studied in 55 isolated Langendorff-perfused rabbit hearts at different pacing cycle lengths (CLs). MAP duration measured at 90% repolarization (APD90) was determined from 6 to 8 endocardial and epicardial MAP recordings with dispersion of ventricular repolarization defined as the range of APD90. The protocol was repeated 60 minutes after initiation of a perfusate containing increasing concentrations of d-sotalol (n = 12, 10[-6] M, 10[-5] M, and 5 x 10[-5] M) and quinidine (n = 8, 10[-6] M and 10[-5] M). Seventeen rabbits were fed with an aqueous solution of amiodarone (50 mg/kg per day over 4 weeks). The data of these experiments (n = 17) were compared with a series of 18 untreated control rabbits. Dispersion of ventricular repolarization was unchanged with the low concentration of d-sotalol (10[-6] M) but was increased-particularly at long CLs-with higher d-sotalol concentrations. With both concentrations of quinidine, dispersion of ventricular repolarization was increased in a rate independent manner. Amiodarone did not affect dispersion of ventricular repolarization. CONCLUSIONS: Rate-dependent and concentration-dependent increases in dispersion of ventricular repolarization by d-sotalol and quinidine in this isolated rabbit heart model may help explain their proarrhythmic effects while the absence of an increase in dispersion of ventricular repolarization with amiodarone correlates with its clinically observed lower incidence of proarrhythmia. PMID- 9395167 TI - Importance of electrode conductive surface area and edge effects on ventricular defibrillation efficacy. AB - INTRODUCTION: The role of edge effects and electrode surface area of the right ventricular (RV) transvenous lead (TVL) on defibrillation efficacy is unknown. METHODS AND RESULTS: Defibrillation threshold (DFT) testing was conducted randomly in 12 dogs using ring electrode leads in an RV/SVC (superior vena cava) or RV/SVC/patch system. The leads (RV-4, RV-8t, RV-8, RV-15) had electrode surface areas of 20%, 20%, 40%, and 70%, respectively. A computer model predicted the magnitude of electrode surface current (RV-8t > RV-4 > RV-8 > RV-15) and the potential distribution (PD) at four sites: electrode surface (site a) and at 2 mm (b), 4 mm (c), and 8 mm (d) away from the surface. Despite different near-field PDs (sites a, b, c), PDs were nearly identical at site d. Resistance decreased as the surface area increased. DFT energy for the RV-15 lead was lower than the RV-4 and RV-8t. There was no difference between energy requirements for the RV-15 and RV-8 leads. No difference was found in DFT current for each lead. Comparison of the RV-8t and RV-4 leads showed no difference in DFT energy despite a lower resistance and a greater number of edges. CONCLUSIONS: Increasing the RV TVL surface area lowered the resistance. However, surface area coverages > or = 40% did not lower DFT energy. No significant change in DFT current occurred despite different predicted near-field current densities. PDs were nearly identical 8 mm from the electrode surface. Thus, the far-field current density appears to play a more important role in determining defibrillation success. PMID- 9395168 TI - Critical atrial site for ablation of pacing-induced atrial fibrillation in the normal dog heart. AB - INTRODUCTION: Radiofrequency catheter ablation (RFA) has been used recently to treat atrial fibrillation (AF). The purpose of this study was to investigate a new approach to preventing AF by RFA. METHODS AND RESULTS: In open chest, anesthetized dogs, AF (lasting > 30 sec) was induced after burst stimulation, and electrophysiologic parameters were recorded before and after RFA. In group 1 (9 dogs) we performed selective and combined slow and fast pathway RFA, whereas in group 2 (11 dogs) RFA was applied as a linear lesion at the mid-atrial septum between the inferior vena cava and the fossa ovalis. After ablation, the Wenckebach cycle length was significantly prolonged only in group 1 (194 +/- 23 vs 282 +/- 35 msec, P = 0.002), whereas the interval between the stimulus (S) artifact applied at the high right atrium to the His bundle (H) (SH interval) prolonged to the same extent in both groups (162 +/- 14 vs 146 +/- 45 msec, P = NS); group 1 due to an A-H prolongation whereas in group 2 it was due to an intra atrial conduction delay. In group 1 AF still remained inducible, although with a longer mean R-R interval (215 +/- 16 vs 433 +/- 88 msec, P < 0.05). No instance of complete AV block developed. In group 2, sustained AF was noninducible in 10 dogs and its duration was markedly shorter in the remaining one (8 sec). Gross anatomy and histology did not reveal any damage inside of Koch's triangle, and particularly to the compact AV node. CONCLUSION: These findings suggest that RFA at the mid-atrial septum prevents AF in the normal dog heart. This approach might also be successful in those clinical settings in which the atrial septum plays a critical role in the maintenance of sustained AF. PMID- 9395169 TI - Ablation of atrial fibrillation. PMID- 9395170 TI - Chronic amiodarone reduces transmural dispersion of repolarization in the canine heart. AB - INTRODUCTION: Amiodarone is a potent antiarrhythmic agent used in the management of both atrial and ventricular arrhythmias. In addition to its beta-blocking properties, amiodarone is known to block the sodium, potassium, and calcium channels in the heart. Its complex electropharmacology notwithstanding, the reasons for the high efficacy of the drug remain unclear. Also not well understood is the basis for the low incidence of proarrhythmia seen with amiodarone relative to other agents with Class III actions. The present study was designed to examine the effects of chronic amiodarone in epicardial, endocardial, and M cells of the canine left ventricle. METHODS AND RESULTS: We used standard microelectrode techniques to record transmembrane activity from endocardial, epicardial, mid-myocardial, and transmural strips isolated from the canine left ventricle. Tissues were obtained from mongrel dogs receiving amiodarone orally (30 to 40 mg/kg per day) for 30 to 45 days or from untreated controls. Chronic amiodarone produced a greater prolongation of action potential duration in epicardium and endocardium, but less of an increase, or even a decrease at slow rates, in the M region, thereby reducing transmural dispersion of repolarization. In addition, chronic amiodarone therapy suppressed the ability of the IKr blocker, d-sotalol, to induce a marked dispersion of repolarization or early afterdepolarization activity. CONCLUSION: Our data demonstrate for the first time a direct effect of chronic amiodarone treatment to differentially alter the cellular electrophysiology of ventricular myocardium so as to produce an important decrease in transmural dispersion of repolarization, especially under conditions in which dispersion is exaggerated. These results may contribute to our understanding of the effectiveness of amiodarone in the treatment of life threatening arrhythmias as well as to our understanding of the low incidence of proarrhythmia attending therapy with chronic amiodarone in comparison with other Class III agents. PMID- 9395172 TI - Retrograde supernormal conduction in concealed accessory atrioventricular pathway following catheter ablation. AB - A case is presented of a 63-year-old woman with a concealed accessory pathway that exhibited retrograde supernormal conduction after radiofrequency catheter ablation. Although ventricular pacing at a slow rate revealed no retrograde conduction over the accessory pathway following ablation, the tachycardia recurred 15 months later. During ventricular pacing there was retrograde 1:1 conduction over the accessory pathway at a fast rate while there was intermittent VA dissociation with rare retrograde conduction at the slower rate. Ventricular extrastimulus testing demonstrated a supernormal conduction zone of the coupling interval. Thus, accessory pathways may exhibit supernormal conduction after catheter ablation. Pacing should be performed at both slow and fast rates to confirm the presence of conduction block following ablation. PMID- 9395171 TI - Effects of sodium channel block with mexiletine to reverse action potential prolongation in in vitro models of the long term QT syndrome. AB - INTRODUCTION: Recent clinical studies have reported a greater effectiveness of sodium channel block with mexiletine to abbreviate the QT interval in patients with the chromosome 3 variant (SCN5A, LQT3) of the long QT syndrome (LQTS) than those with the chromosome 7 form of the disease (HERG, LQT2), suggesting the possibility of gene-specific therapy for the two distinct forms of the congenital LQTS. Experimental studies using the arterially perfused left ventricular wedge preparation have confirmed these clinical observations on the QT interval but have gone on to further demonstrate a potent effect of mexiletine to reduce dispersion of repolarization and prevent torsades de pointes (TdP) in both LQT2 and LQT3 models. A differential action of sodium channel block on the three ventricular cell types is thought to mediate these actions of mexiletine. This study provides a test of this hypothesis by examining the effects of mexiletine in isolated canine ventricular epicardial, endocardial, and M region tissues under conditions that mimic the SCN5A and HERG gene defects. METHODS AND RESULTS: We used standard microelectrode techniques to record transmembrane activity from endocardial, epicardial, mid-myocardial, and transmural strips isolated from the canine left ventricle. d-Sotalol, an IKr blocker, was used to mimic the HERG defect (LQT2), and ATX-II, which increases late Na channel current, was used to mimic the SCN5A defect (LQT3). d-Sotalol (100 microM) preferentially prolonged the action potential of the mid-myocardial M cell (APD90 increased from 340 +/- 65 to 623 +/- 203 msec) as did ATX-II (10 to 20 nM; APD90 increased from 325 +/- 51 to 580 +/- 178 msec; basic cycle length = 2000 msec), thus causing a marked increase in transmural dispersion of repolarization (TDR). Mexiletine (2 to 20 microM) dose-dependently reversed the ATX-II-induced prolongation of APD90 in all three cell types. Mexiletine also reversed the d-sotalol-induced prolongation of the M cell action potential duration (APD), but had little effect on the action potential of epicardium and endocardium. Due to its preferential effect to abbreviate the action potential of M cells, mexiletine reduced the dispersion of repolarization in both models. Low concentrations of mexiletine (5 to 10 microM) totally suppressed early afterdepolarization (EAD) and EAD-induced triggered activity in both models. CONCLUSIONS: Our results indicate that the actions of mexiletine are both cell and model specific, but that sodium channel block with mexiletine is effective in reducing transmural differences in APD and in abolishing triggered activity induced by d-sotalol and ATX-II. The data suggest that mexiletine's actions to reduce TDR and prevent the induction of spontaneous and programmed stimulation-induced TdP in these models are due to a preferential effect of the drug to abbreviate the APD of the M cell and to suppress the development of EADs. The data provide further support for the hypothesis that block of the late sodium current may be of value in the treatment of LQT2 as well as LQT3 and perhaps other congenital and acquired (drug-induced) forms of LQTS. PMID- 9395173 TI - A case of widely split double P waves with marked intra-atrial conduction delay. AB - We report a 78-year-old man as the first documented case of double P waves separated by 400 msec on 12-lead ECG. These P waves had different polarities on lead V1. The first P wave represented activation of the lateral wall of the right atrium, and the latter P wave represented activation of the medial right atrium and the left atrium. Widely spaced double potentials were recorded craniocaudally along the line, presumably corresponding to the crista terminalis during sinus rhythm. For this to occur, conduction disturbance has to be present both in the upper and lower right atrium. Conduction disturbance in the upper right atrium would interrupt excitation from the sinus node to the medial wall, and conduction disturbance in the lower right atrium would interrupt excitation spreading from the lower lateral right atrium to the isthmus area where fragmented potentials were recorded. These multiple discrete lesions appear to constitute a unique electrical atriopathy in this patient. PMID- 9395174 TI - Atypical atrioventricular nodal reentry tachycardia with atrioventricular block mimicking atrial tachycardia: electrophysiologic properties and radiofrequency ablation therapy. AB - INTRODUCTION: Fast-intermediate form AV nodal reentry tachycardia (AVNRT) sometimes may mimic atrial tachycardia or atrial flutter and render the diagnosis difficult when the tachycardia rate is fast and AV block occurs during tachycardia. METHODS AND RESULTS: A 45-year-old woman with paroxysmal supraventricular tachycardia was referred to this institution. Initially, the tachycardia was thought to be an atrial tachycardia because of: (1) a short cycle length of the tachycardia with 2:1 and Wenckebach AV block; (2) a difference in the atrial activation sequence during tachycardia and during ventricular pacing; and (3) failure of burst ventricular pacing to affect the atrial rate and the atrial activation sequence during tachycardia. An accurate diagnosis of fast intermediate form AVNRT was subsequently made based on the finding that the tachycardia was induced following delivery of a third ventricular extrastimulus, which showed a sequence of V-A-H and a change on atrial activation sequence of the induced beat. Successful radiofrequency ablation was achieved only after accurate diagnosis of the tachycardia was made. CONCLUSION: Fast-intermediate form AVNRT sometimes may masquerade as atrial tachycardia. Accurate diagnosis is mandatory for successful ablation therapy. PMID- 9395175 TI - Radiofrequency catheter ablation of ventricular tachycardia late after myocardial infarction. AB - Radiofrequency catheter ablation is a promising method for controlling ventricular tachycardia (VT) due to prior myocardial infarction. Limitations of mapping and ablation techniques have largely restricted its use to selected patients who have hemodynamically tolerated sustained monomorphic VT that allows catheter mapping. Multiple monomorphologies of VT, which are usually present, often complicate the ablation procedure and interpretation of ablation effects. Ablation is generally restricted to experienced centers and is usually reserved for patients who have failed other therapies. Despite these difficulties, successful ablation can be life-saving in patients with incessant VT and can markedly improve quality of life with frequent shocks from implantable defibrillators. PMID- 9395176 TI - Management of the child with Wolff-Parkinson-White syndrome and supraventricular tachycardia: model for cost effectiveness. AB - In the next decade, "better" management will be defined by cost effectiveness including morbidity, mortality, and cost. We used a cost-effectiveness model for children with Wolff-Parkinson-White syndrome (WPW) and supraventricular tachycardia (SVT) comparing medical, surgical, and catheter ablative treatment between age 5 years (estimated average age at first recurrence after infancy) and age 21. Charges were quantitated from actual hospital bills; mortality was estimated from the literature; morbidity was assessed by estimating the number of hours in SVT, hours in clinic, hours in routine hospital bed, and hours in hospital intensive care; and the hours were then multiplied by a severity factor, normalized to 1.0 for 1 hour of SVT (0.5 for 1 hour in clinic, 0.75 for routine hospital, and 2.0 for intensive care). Overall charges (5 to 21 years old) for catheter ablation ($17,236) were 39% of surgical management and 57% of medical management; estimated mortality for catheter ablation (5 to 21 years old including failures that reverted to medical management) was 0.15%, which was 10% of medical management and 28% of surgical management; morbidity for catheter ablation was 27.6 units, which was 32% of medical management and 36% of surgical management. Sensitivity analysis demonstrated that the catheter ablation strategy remained preferable throughout the range of plausible values of cost, mortality, and morbidity (including a repeat procedure for initial failures). Therefore, catheter ablation has lower cost, mortality, and morbidity than either medical management or surgery and is the treatment of choice for the child 5 years of age or older with WPW and SVT. This type of analysis can be used for other forms of chronic disease in children. PMID- 9395177 TI - Wide complex tachycardia in a critically ill patient: what is the rhythm? PMID- 9395178 TI - Electrophysiology of the atrio-AV nodal inputs and exits in the normal dog heart: radiofrequency ablation using an epicardial approach. PMID- 9395180 TI - Detection of BCL-6 rearrangements and p53 mutations in Malt-lymphomas. AB - Twenty-seven lymphomas of mucosa-associated lymphoid tissue (MALT) derived from distinct anatomical sites were tested for the presence of genetic lesions commonly involved in B-cell lymphomagenesis, including activation of proto oncogenes (BCL-1, BCL-2, BCL-6, and c-MYC), disruption of tumor suppressor loci (p53, 6q), and infection by viruses [Epstein-Barr virus (EBV), and Kaposi's sarcoma-herpesvirus/human herpesvirus-8 (KSHV/HHV-8)]. Sixteen low-grade and 11 high-grade MALT-lymphomas were included in the study. The presence of genetic lesions was tested by a combination of molecular approaches, including Southern blot hybridization, polymerase chain reaction (PCR), and PCR-single strand conformation polymorphism followed by DNA direct sequencing. Alterations of BCL 1, BCL-2, or c-MYC, as well as infection by KSHV/HHV-8, scored negative in all MALT-lymphomas analysed. Conversely, rearrangements of BCL-6 and mutations of p53 clustered with a fraction of high-grade MALT-lymphomas. Deletions of 6q occurred in selected cases of both low- and high-grade MALT-lymphomas, whereas a monoclonal infection by EBV was restricted to one single patient. These data corroborate the notion that the molecular pathogenesis of MALT-lymphomas differs substantially from that of nodal B-cell lymphomas. Occasionally, however, a proportion of high-grade MALT-lymphomas may harbor selected genetic lesions among the ones commonly involved in nodal B-cell lymphomagenesis. PMID- 9395179 TI - Cell behavior and signal molecule involvement in a case study of malignant histiocytosis: a negative model of morphine as an immunoregulator. AB - In a patient diagnosed with histiocytic medullary reticulosis (HM), we examined immunocytes for their responsiveness towards known signaling molecules. Both the granulocytes and monocytes were found to exhibit a high level of spontaneous activation (96% compared to normal cells 7%; P < 0.001). These cells could not be downregulated when exposed to morphine. Following patient treatment with doxorubicin and cyclophosphamide, the immunocytes still exhibited a high spontaneous activation. They responded to morphine exposure in vitro with a cell rounding and becoming immobile for only 20 min whereas normal cells would remain round and immobile for up to 1-2 h. An examination of the plasma from the HM patient revealed that monocyte colony stimulating factor (MCSF) levels were elevated (6.4 and 5.78 compared to a control range of 1-1.75 ng/ml). In the HM patient, the immunocytes did not express the opiate selective and opioid peptide insensitive receptor, mu3, supporting the lack of opiate action. Given this finding, we incubated normal monocytes with MCSF and found that it significantly reduced the mu3 Bmax. Given the role of intracellular calcium in the activation process of immunocytes, we examined the action of various calcium channel blockers for their ability to inhibit the activated HM monocytes. The agents (nimodipine, cardiazem, and verapamil; 10[-9] M) were able to inhibit the HM associated chemokinesis. Taken together, the data indicate that in the HM patient the immunocytes appear to be overactivated because stimulatory molecules are high and have the ability to downregulate the normal "braking" process. Additionally, the data indicate that MCSF deregulation may be involved as an initiating factor for this disorder. PMID- 9395181 TI - Sequential mitoxantrone, daunorubicin, and cytosine arabinoside for patients with newly diagnosed acute myelocytic leukemia. AB - Mitoxantrone (M) is a synthetic aminoanthraquinone with anti-leukemic activity in patients with daunorubicin (D) resistant acute leukemia. The Cancer and Leukemia Group B (CALGB) has undertaken a limited access pilot study in which M, 12 mg/m2, over 30 min, daily for 3 days, and cytosine arabinoside (Ara-C), 100 mg/m2/day by constant infusion for 7 days were used for the induction of newly diagnosed patients with AML. Responding patients were consolidated with daunorubicin, 45 mg/m2/day for 3 days, and 7 days of Ara-C. After a second consolidation identical to induction, no further therapy was given. Twenty-nine patients with a median age of 50 years (range 18-72) were entered in the study; 18 were males and 11 females. Twenty-four (83%) patients achieved CR, 1 patient achieved a PR, and 4 died in induction from leukemia-related causes. Two patients died in CR from consolidation-related neutropenic sepsis and two additional patients died in CR. Of 24 patients, 7 remain disease-free at a median follow-up interval of 8 years. The regimen is active and well tolerated. The duration of disease-free survival in responding patients is consistent with that seen in similar regimens using intensification chemotherapy without prolonged maintenance. PMID- 9395182 TI - Lymphoma with multi gene rearrangement on the level of immunoglobulin heavy chain, light chains, and T-cell receptor beta chain. AB - A unique case with diffuse mixed malignant lymphoma was investigated for gene rearrangement on the level of T-cell receptor (TCR), heavy chain immunoglobulin (Ig), and both light chains. Cell phenotype was examined with immunofluorescence techniques using antibodies against surface immunoglobulins (SIg) and the kappa and lambda light chains. Monoclonal antibodies were used against CD3, CD4, CD5, CD8, CD10, CD19, CD22, HLA-DR, and TdT. Gene rearrangement analysis for monoclonality determination was carried out with restricted DNA (EcoR I and Hind III) hybridized with one of the following 32P-labelled probes: T-cell receptor (TCR beta), immunoglobulin heavy chain (JH), k light chain, and lambda light chain. Phenotyping of the cell population from the excised lymph node (LN) revealed the presence of 66% B-cells and 35% T-cells. Most of the B cells (94%) expressed mu heavy chain only. Expression of both light chains was negligible (k = 7% and lambda = 2%). Gene rearrangement, which indicates monoclonality, was positive on the level of TCR, Ig heavy chain, and both light chains. The data obtained suggests a neoplastic transforming event in lymphoid stem cells, which preceded the subsequent differentiation process into either B or T lymphoma. PMID- 9395183 TI - MDS and AML with trisomy 8 as the sole chromosome aberration show different sex ratios and prognostic profiles: a study of 115 published cases. AB - A number of chromosome aberrations occur nonrandomly as the sole aberration in malignant and premalignant hematological disorders. They imply very different prognoses. For most of them the survival consequences have been established. For trisomy 8, which is the most frequent numerical aberration in myeloid disorders, the prognostic implications have not been investigated. In order to clarify survival in patients with trisomy 8 as the sole aberration, the literature was searched for such cases. In 115 patients survival data were available. In 103 (89.6%), a myeloid disorder had been diagnosed. Acute myeloid leukemia (AML) or a myelodysplastic syndrome (MDS) occurred in 100 cases (87.0%). The median survival was found to be 17.1 months. On multivariate survival analysis (Cox), age above 60 and a leukemic diagnosis were found to be independent adverse prognostic indicators. MDS patients survived significantly longer (median 21 months) than AML patients (median 15 months). In MDS age and in AML the trisomy 8 clonal size was an independent prognostic factor. An unexpected observation was a clear male preponderance in trisomy 8 MDS (about two-thirds of cases). In trisomy 8 AML an approximate 1:1 ratio was found. Browsing of Mitelman's catalog confirmed these ratios. PMID- 9395184 TI - Serum methylmalonic acid and total homocysteine in patients with suspected cobalamin deficiency: a clinical study based on gastrointestinal histopathological findings. AB - We compared the sensitivity and specificity of the two metabolite tests, methylmalonic acid (MMA) and total homocysteine (Hcy) in serum, and serum cobalamin (Cbl) in patients referred to our hospital because of suspected cobalamin deficiency and a serum cobalamin value at the referring unit <200 pmol/L. All 111 patients included were investigated using upper gastrointestinal endoscopy with biopsy specimens taken from the gastric and duodenal mucosa to find a morphological basis for cobalamin malabsorption as well as the Schilling test for the validation of the serum tests. All patients were treated with cobalamin and new blood samples were taken after 4 weeks. We found no difference in sensitivity and specificity between serum MMA, Hcy, and Cbl in identifying patients with and without conditions compatible with cobalamin malabsorption. Elevated serum MMA and Hcy were also found in about 15% of the group of patients with normal Schilling tests and without a morphological basis for cobalamin malabsorption. Moreover, most patients in this group responded with decreased values of the metabolite tests following cobalamin treatment, suggesting that neither elevated metabolites nor a decrease in these values following cobalamin treatment are specific for cobalamin deficiency. PMID- 9395185 TI - Dissection of the association status of two polymorphisms in the beta-globin gene cluster with variations in F-cell number in non-anemic individuals. AB - Expression of fetal hemoglobin (Hb F) is under polygenic control involving determinants both linked and unlinked to the beta-globin gene cluster on chromosome 11. Variations in the DNase I-hypersensitive site 2 of the locus control region (LCR-HS2) and a C --> T change at position -158 from the Ggamma gene (detected as an XmnI polymorphism) correlate with the high level of Hb F expression in patients with sickle-cell anemia and beta-thalassemia. Interpretation of data under these conditions of anemic stress is difficult because the preferential survival of Hb F-containing erythrocytes (F-cells) may not reflect the true status of Hb F expression. We investigated the relationship between these markers and Hb F expression in terms of F-cell levels in 48 unrelated non-anemic AS heterozygotes from Sicily. The betaS-chromosome of all these individuals was of the Benin haplotype and they differed only by their betaA chromosomes. We demonstrate that F-cell expression is more strongly associated with LCR-HS2 polymorphism than with XmnI polymorphism. The observed association between XmnI polymorphism and Hb F expression is very likely to be due to linkage disequilibrium with LCR-HS2 sequences. PMID- 9395186 TI - Effects of hyperthermal stress on the ultrastructure of platelets with reference to the localization of platelet peroxidase and fibrinogen in vivo. AB - Ultrastructure of platelets with the localization of platelet peroxidase and fibrinogen through 3-min 47 degrees C hot-spring bathing was investigated in eight healthy volunteers. The mean sublingual temperature rose about 1.8 degrees C 5 min after the start of bathing. The frequencies of fold, pseudopods, vacuoles, and centralization were increased after bathing. Platelet peroxidase activity was decreased after bathing. Furthermore, fibrinogen was decreased in alpha-granules after bathing. Thus, hyperthermal stress in vivo may activate platelets, resulting in consumption of platelet peroxidase and fibrinogen. PMID- 9395187 TI - Parvovirus B19 quiescence during the course of human immunodeficiency virus infection in persons with hemophilia. AB - To detect and characterize parvovirus B19 infection during the course of progressive immune deficiency from human immunodeficiency virus (HIV), ten subjects enrolled in the Multicenter Hemophilia Cohort Study were followed for 6.4 to 15 years from HIV seroconversion through extreme immune deficiency. Four to five sera or plasma samples from each subject, collected at predetermined CD4+ lymphocyte levels, were tested for immunoglobulin G (IgG) and M (IgM) B19 antibodies and DNA. All 42 samples were positive for B19 IgG antibodies, and three were weakly positive for IgM antibodies. Only one sample, collected coincident with HIV seroconversion, was unequivocally positive for B19 DNA. No persistent hematologic adverse effects of B19 infection were observed. Thus, although B19 IgG antibodies are highly prevalent among HIV-infected persons with hemophilia or related disorders, B19 viremia and its hematologic consequences were not detected, even with severe depletion of CD4+ lymphocytes. If primary B19 infection occurs after immune deficiency, however, the consequences may be more adverse. PMID- 9395188 TI - BFU-E colony growth in response to hydroxyurea: correlation between in vitro and in vivo fetal hemoglobin induction. AB - Patients with sickle-cell anemia treated with hydroxyurea may have significant reduction in frequency and severity of pain episodes. However, previous clinical trials show a variable response to hydroxyurea. Criteria which can be used to select patients who are likely to respond to hydroxyurea treatment would be useful. Our laboratory has previously demonstrated an inverse linear relationship between the total number of burst-forming unit-erythroid (BFU-E) colonies and fetal hemoglobin levels in sickle-cell patients treated with hydroxyurea. In the present report, an in vitro cell culture system was established to evaluate the effects of hydroxyurea on BFU-E colony growth and induction of fetal hemoglobin production. Five Hb SS patients who were not previously treated with hydroxyurea and three Hb SS patients who failed to respond to hydroxyurea treatment were included in the study. The results show that the number of BFU-E colonies is decreased from 153.7 to 7.2 per 3 x 10(5) mononuclear cells, whereas fetal hemoglobin levels were increased from 5.1 to 19.4% in the presence of hydroxyurea in vitro in cultured erythroid progenitors, which were derived from 5 patients before treatment. The number of BFU-E colonies decreased from 153.7 to 2.0 per 3 x 10(5) mononuclear cells in the in vitro cultures obtained from serial peripheral blood samples over a 9- to 20-week period of oral hydroxyurea therapy. A simultaneous rise in fetal hemoglobin level from 10.2 to 28.6% in the peripheral blood over the same period of hydroxyurea therapy was also observed. Our results demonstrate that the increase in fetal hemoglobin levels in cells treated with hydroxyurea in vitro is comparable to the rise of fetal hemoglobin production following hydroxyurea therapy in these patients. On the contrary, these findings were not observed in three previously non-responsive sickle-cell patients. These results suggest that the changes in number of BFU-E colonies and fetal hemoglobin levels after in vitro exposure to hydroxyurea may be a useful approach to select sickle-cell patients who will respond to hydroxyurea therapy. PMID- 9395189 TI - Release of adenosine triphosphate by adenosine diphosphate in whole blood and in erythrocyte suspensions. AB - In whole blood samples from thrombocytopenic patients, large amounts of ATP were released by ADP, exceeding the level obtained with samples from normal persons by far. Because we suspected that the high potential of ATP in erythrocytes would be the main source for this phenomenon, the release of ATP by ADP was measured in whole blood samples from normal, thrombocytopenic, and leukocytopenic persons and in suspensions of washed erythrocytes. The release was recorded by a Whole Blood Lumi-Aggregometer type 500 VS (Chrono-Log Corporation, Havertown, PA) using the luciferin-luciferase system. Not only in samples from thrombocytopenic persons but also with normal platelet count, increasing amounts of ATP were released with increasing ADP concentrations, finally exceeding the ATP releasable from thrombocytes by thrombin. The amounts of ADP required to match the ATP release of thrombin were closely correlated with the platelet counts in the samples. With lower platelet counts, the release mechanism from erythrocytes could be stimulated more easily by low concentrations of ADP. The binding of ADP to platelets occurred with ostensibly higher affinity. The phenomenon of overshooting ATP release was also observed in samples from extremely leukocytopenic patients. A very large release of ATP was also achieved in suspensions of washed erythrocytes. In this way our hypothesis of ATP release from erythrocytes by ADP was confirmed again. The mechanism of the release from erythrocytes remains unclear. We speculate that its purpose is to regulate extracellular nucleotides in the circulating blood. PMID- 9395191 TI - Cyclic thrombocytopenia in a patient treated with cyclosporine for refractory idiopathic thrombocytopenic purpura. AB - Cyclic thrombocytopenia (CT) is a rare disorder with cyclic changes of the platelet counts. Though the pathogenesis of this disorder has not been clarified, recent reports suggest that periodic destruction and/or ineffective production of platelets may be important causes of the disease. We report a case of a patient with refractory idiopathic thrombocytopenic purpura (ITP) in whom CT developed after cyclosporine A (CyA) therapy. There was an inverse relation between platelet counts and the serum levels of platelet-associated immunoglobulin G (PAIgG). The ploidy of bone marrow megakaryocytes also had an inverse relation with platelet counts. When the platelet count was low, the ploidy of megakaryocytes increased (P < 0.01). The number and area of bone marrow megakaryocytes were unrelated to platelet counts. These results indicate the possibility of platelet destruction caused by immunological mechanisms in CT. Cyclosporine A could have certain but fluctuating regulatory effects against antibody production for circulating platelets, which could lead to cyclic changes of the platelet counts. This case also suggests that CyA can be effective in severe refractory ITP. Regulatory mechanisms of platelet production and destruction and appropriate doses of CyA should be further studied in autoimmune mediated thrombocytopenias. PMID- 9395190 TI - Telomere length in myelodysplastic syndromes. AB - We have studied telomere length in the bone marrow cells or the granulocyte and lymphocyte cell fractions of 54 patients with myelodysplastic syndromes (MDS) by Southern blot hybridization using the (TTAGGG)4 probe. The average telomere length expressed as the peak telomere repeat array (TRA) in the peripheral blood, or bone marrow samples obtained from a group of 21 healthy age-matched controls (26-89 years old, mean age 55), ranged between 7.5 and 9.5 kb (mean peak TRA 8.6 kb). Twenty-four patients with refractory anemia (RA) were studied; 10/24 (42%) had telomere reduction (<7.5 kb) relative to age-matched controls and the mean peak TRA was 7.5 kb (range 4.0-9.0 kb). Eleven patients with RA with excess blasts (RAEB) were studied; 5/11 (45%) had reduced telomeres relative to age matched controls and the mean peak TRA was 7.1 kb (range 5.0-9.0 kb). Eighteen patients with MDS in transformation to AML, comprising 15 with RAEB in transformation (RAEBt) and 3 with CMML in transformation (CMMLt), were also studied. Thirteen of eighteen patients (72%) had telomere reduction relative to age-matched controls and the mean peak TRA was 6.1 kb (range 3.5-9.0 kb). Thirty six patients included in the study had either a normal karyotype or a simple karyotype (1 karyotypic change) and 20/36 (55%) of these had telomere reduction and the mean peak TRA was 7.1 kb (range 4.3-9.0 kb); 8 patients had a complex karyotype (3 or more karyotypic changes) and 5/8 (62%) of these had telomere reduction and the mean peak TRA was 6.1 kb (range 3.5-9.0 kb). We conclude, firstly that there is heterogeneity of telomere length in MDS and that this is observed throughout the spectrum of FAB-subtypes. Secondly, these data show that a marked reduction in telomere length in MDS if often associated with leukemic transformation and with the presence of complex karyotypic abnormalities. PMID- 9395192 TI - Intra-mesenteric artery steroid administration relieved severe refractory gastro intestinal graft-vs.-host disease in an allogeneic bone marrow transplantation patient. AB - We report a case of severe gastro-intestinal (G-I) graft-vs.-host disease (GVHD) successfully treated with intra-mesenteric artery steroid administration. A 29 year-old man with severe aplastic anemia (SAA) was submitted to HLA-identical unrelated allogeneic bone marrow transplantation (BMT) and was found to be suffering from grade IV G-I GVHD. Although cyclosporine, steroid pulse therapy, and FK506 proved ineffective, 30 mg of water-soluble prednisolone as administered into each the superior and inferior mesenteric artery with remarkable effects. This treatment was repeated two times, and the symptoms of G-I GVHD disappeared completely. PMID- 9395193 TI - Concomitant chronic lymphocytic leukemia and acute myeloid leukemia with an uncommon immunophenotype. AB - We report a case of simultaneous diagnosis of chronic lymphocytic leukemia (CLL) and acute myeloid leukemia (AML), in which the use of flow cytometry analysis allowed the demonstration of two different cell populations and the study of both immunophenotyping patterns with a large panel of monoclonal antibodies (MoAbs). CLL cells showed a typical immunophenotype with coexpression of B cell markers with CD5, CD23, CD43, and weak surface immunoglobulin light chain restriction expression, whereas the AML population had a very uncommon phenotype with expression of myeloid markers and CD56 and lack of expression of other natural killer (NK) antigens, CD34 and HLA-DR. After chemotherapeutic treatment of AML with two induction courses, the patient achieved complete remission of the AML with persistence of a CD19/CD5 positive population. After consolidation chemotherapy, this latter population was no longer detectable despite the presence of lymphoid nodules in a bone marrow biopsy. Six months after diagnosis, the patient relapsed with AML and died shortly afterwards. PMID- 9395194 TI - Multiple myeloma associated with diffuse osteosclerotic bone lesions: a clinical entity distinct from osteosclerotic myeloma (POEMS syndrome). AB - Multiple myeloma usually is characterized by the development of lytic bone lesions. Osteosclerotic myeloma is a rare entity characterized by a single or multiple osteosclerotic bone lesions and often accompanied by a demyelinating polyneuropathy. Multiple myeloma associated with widespread osteosclerotic lesions seen on radiographic studies is exceedingly rare. We describe 3 such cases and review 12 other cases described in the literature. Overall, the patients described herein had a clinical course that resembled multiple myeloma more than osteosclerotic myeloma. However, some patients had features of both diseases. Although rare, multiple myeloma should be included in the differential diagnosis of widespread osteosclerotic bone lesions. PMID- 9395195 TI - Acquired dysfibrinogenemia following allogeneic bone marrow transplantation. PMID- 9395196 TI - Identification of a myeloma variant with aggressive biological and clinical characteristics: "early" plasma cell meningitis. PMID- 9395197 TI - Microparticles and coronary artery disease. PMID- 9395198 TI - Role of PPAR alpha in the mechanism of action of the nongenotoxic carcinogen and peroxisome proliferator Wy-14,643. AB - Chronic administration of peroxisome proliferators to mice and rats results in hepatomegaly and ultimately carcinogenesis. The mechanism underlying the carcinogenic effect of nongenotoxic peroxisome proliferators is not well understood. To determine whether nongenotoxic carcinogenesis is receptor mediated, we evaluated the effect of the prototypical peroxisome proliferator Wy 14,643 on replicative DNA synthesis and carcinogenesis in the PPAR alpha-null mouse line. Male mice (F4, Sv/129 ter) of both genotypes (+/+) and (-/-) were fed either a control diet or one containing 0.1% Wy-14,643 for either 1 week, 5 weeks, or 11 months. Wild-type mice fed the Wy-14,643 diet for 1 or 5 weeks showed increased hepatic labeling by bromodeoxyuridine (BrDU) compared to untreated controls. In contrast, there was no increase in hepatic BrDU labeling index in (-/-) mice fed the Wy-14,643 diet for the same time periods compared to controls. After 11 months, 100% of the (+/+) mice fed the Wy-14,643 diet had multiple hepatocellular neoplasms, including adenomas and carcinomas, while the ( /-) mice fed the Wy-14,643 diet were unaffected. This work demonstrates that the effects of Wy-14,643 on replicative DNA synthesis and hepatocarcinogenesis are mediated by PPAR alpha. PMID- 9395199 TI - The carcinogenic potential of the gas phase of environmental tobacco smoke. AB - Female strain A/J mice were exposed to unfiltered or HEPA-filtered environmental tobacco smoke (ETS). Total suspended particulates (TSP) in the full smoke exposure chamber was 78.5 mg/m3 and in the filtered smoke chamber 0.1 mg/m3; nicotine concentrations in the full and filtered smoke chamber were 13.4 and 3.1 mg/m3, respectively. Animals exposed to filtered ETS (6 h a day, 5 days a week) and killed after 5 months had a higher lung tumor incidence and multiplicity than controls maintained in filtered air, although the differences were not statistically significant. Animals exposed to filtered and full ETS and allowed to recover in air for 4 months had an average of 1.2 +/- 0.3 tumors per lung and 1.3 +/- 0.3 tumors per lung, respectively. Air exposed control animals had an average tumor multiplicity 0.5 +/- 0.1 tumors per lung. Increased immunostaining for CYP 1A1 was not evident in the lung of animals exposed to filtered smoke. Based on the chamber concentrations of selected nitrosamines and polycyclic aromatic hydrocarbons, the possible maximum uptakes by the mice of NNK, NNN and benzo[a]pyrene during the 5 months exposure period were three to six orders of magnitude below doses reported in the literature to produce 1 lung tumor in strain A/J mice. It was concluded that the gas phase of ETS is as carcinogenic as is full ETS. The carcinogenicity of the gas phase may be due to some as yet unidentified, yet highly potent carcinogens or by placing a substantial, possibly free radical-mediated oxidative stress on the lung. PMID- 9395200 TI - Emergence of undifferentiated rat tracheal cell carcinomas, but not squamous cell carcinomas, is associated with a loss of expression of E-cadherin and of gap junction communication. AB - A series of cells representing normal, non-tumorigenic cell lines, as well as differentiating neoplastic and undifferentiated neoplastic rat tracheal epithelial cell populations were evaluated for their ability to establish homologous and/or heterologous cell-cell gap junction communication in culture. Gap junction communication was evaluated by flow cytometric quantitation of the transfer of the fluorescent dye calcein from a donor to a recipient cell population via gap junctions. The data indicate that normal primary cultures of rat tracheal epithelial cells, as well as non-tumorigenic cell lines and squamous cell carcinomas cell populations, retain the ability to establish both homologous and heterologous gap junction communication. In all cases an average of >48% of recipient cells had acquired calcein label during a 5-h interval of co-culture of donor and recipient cells at confluent densities. Cells harvested directly from squamous cell carcinoma tumors exhibited similar levels of cell-cell communication. In contrast, cells giving rise to undifferentiated carcinomas, as well as cells harvested from undifferentiated carcinomas, exhibited very low levels or no homologous or heterologous cell-cell communication. Cell populations exhibiting distinctly different communication phenotypes were evaluated by Northern blot analysis for expression of connexins (Cx 26, 32 and 43) and E cadherin. Neither communicating nor non-communicating cells expressed connexin 32. Those cell populations, which established functional gap junctions, expressed E-cadherin as well as connexin 26 and/or 43. In contrast, those cell populations that lacked the ability to communicate universally lacked expression of E cadherin, and a quarter also lacked expression of detectable levels of connexin. PMID- 9395201 TI - Mutagenicity of oxidative DNA damage in Chinese hamster V79 cells. AB - We have investigated the mutagenicity of oxidative DNA damage induced in V79 Chinese hamster lung fibroblast, and measured 8-hydroxydeoxyguanosine (8OHdG) levels as an indicator of this damage. A hydroxyl radical generator, N,N'-bis(2 hydroxyperoxy-2-methoxyethyl)-1,4,5,8-naphthalene-tetra -carboxylic-diimide (NP III), induced 8OHdG in V79 upon irradiation with 366 nm ultraviolet light (UV) for 15 min. 8OHdG was determined by HPLC with electrochemical detection after anaerobic sample processing. The 8OHdG level in the cells treated without NP-III was 0.49 per 10(5) dG, whereas levels in the cells treated with 5, 10 or 20 microM NP-III and UV irradiation were 1.84, 4.06 or 6.95 per 10(5) dG, respectively. The 8OHdG induced by 20 microM NP-III with UV irradiation decreased rapidly, and the half-life of the induced 8OHdG was approximately 6 h. NP-III with UV irradiation also induced DNA strand breaks in all cells uniformly, as determined by single cell gel assay. Mutant frequencies at the hypoxanthine guanine phosphoribosyltransferase (hprt) locus in V79 were determined as the number of 6-thioguanine-resistant cells per 10(6) cells. Mutant frequency of the cells without NP-III was 8.0, and frequencies of the cells treated with 5, 10 or 20 microM NP-III and UV irradiation were 14.9, 20.6 or 24.7 respectively. Treatment with 20 microM NP-III and UV irradiation decreased the cell number, determined 3 days after the treatment, to 20.8%. These findings indicate that acutely induced oxidative DNA damage including mutagenic 8OHdG is only weakly mutagenic in V79. PMID- 9395202 TI - Allelotype and replication error phenotype of small cell lung carcinoma. AB - Allelotype and replication error (RER) phenotype analyses were performed to clarify the pathogenetic significance of inactivation of tumor suppressor genes and genomic instability in the genesis and progression of small cell lung carcinoma (SCLC). We examined 37 cases of SCLC for loss of heterozygosity (LOH) and microsatellite instability at 49 loci on all 39 nonacrocentric chromosomal arms. LOH was frequently (>70%) detected on chromosomes 3p (29/32, 90.6%), 5q (15/21, 71.4%), 13q (25/26, 96.2%), 17p (22/25, 88.0%), and 22q (24/33, 72.7%). Frequent LOH (>70%) on these loci was observed even among seven cases of stage I tumors. The incidence of LOH on all 39 nonacrocentric chromosomal arms was not significantly different between primary tumors and metastases. These results suggest that inactivation of multiple tumor suppressor genes accumulates relatively early during progression of SCLC and it may be responsible for clinically and biologically aggressive phenotype of SCLC. RER was observed in 6/37 (16.2%) of SCLC, however, RER at multiple loci was observed only in two cases. Therefore, it was indicated that genomic instability is uncommon, but might play a role in the genesis of a small subset of SCLC. PMID- 9395203 TI - Stable overexpression of PML alters regulation of cell cycle progression in HeLa cells. AB - Our previous studies demonstrated that PML is a growth suppressor that suppresses oncogenic transformation of NIH/3T3 cells and rat embryo fibroblasts. PML is a nuclear matrix-associated phosphoprotein whose expression is regulated during the cell cycle. Disruption of PML function by t(15;17) in acute promyelocytic leukemia (APL) plays a critical role in leukemogenesis. To further study the role of PML in the control of cell growth, we have stably overexpressed PML protein in the HeLa cell line. This overexpression of PML significantly reduced the growth rate of HeLa cells and suppressed anchorage-independent growth in soft agar. We consequently investigated several parameters correlated with cell growth and cell cycle progression. We found that, in comparison with the parental HeLa cells, HeLa/PML stable clones showed proportionally more cells in G1 phase, fewer cells in S phase and about the same number in G2/M phase. The HeLa/PML clones showed a significantly longer doubling time as a result of a lengthening of the G1 phase. No effect on apoptosis was found in HeLa cells overexpressing PML. This observation indicates that PML suppresses cell growth by increasing cell cycle duration as a result of G1 elongation. To further understand the mechanism of the effect of PML on HeLa cells, expression of cell cycle-related proteins in HeLa/PML and parental HeLa cells was analyzed. We found that Rb phosphorylation was significantly reduced in PML stable clones. Expression of cyclin E, Cdk2 and p27 proteins was also significantly reduced. These studies indicate that PML affects cell cycle progression by mediating expression of several key proteins that normally control cell cycle progression. These results further extend our current understanding of PML function in human cells and its important role in cell cycle regulation. PMID- 9395204 TI - Presence and consequence of uracil in preneoplastic DNA from folate/methyl deficient rats. AB - Uracil can arise in DNA by misincorporation of dUTP into nascent DNA and/or by cytosine deamination in established DNA. Based on recent findings, both pathways appear to be promoted in the methyl-deficient model of hepatocarcinogenesis. A chronic increase in the ratio dUTP:dTTP with folate/methyl deficiency can result in a futile cycle of excision and reiterative uracil misincorporation leading to premutagenic apyrimidinic (AP) sites, DNA strand breaks, DNA fragmentation and apoptotic cell death. The progressive accumulation of unmethylated cytosines with chronic methyl deficiency will increase the potential for cytosine deamination to uracil and further stress uracil mismatch repair mechanisms. Uracil is removed by a highly specific uracil-DNA glycosylase (UDG) leaving an AP site that is subsequently repaired by sequential action of AP endonuclease, 5' phosphodiesterase, a DNA polymerase and DNA ligase. Since the DNA polymerases cannot distinguish between dUTP and dTTP, an increase in dUTP:dTTP ratio will promote uracil misincorporation during both DNA replication and repair synthesis. The misincorporation of uracil for thymine (5-methyluracil) may constitute a genetically significant form of DNA hypomethylation distinct from cytosine hypomethylation. In the present study a significant increase in the level of uracil in liver DNA as early as 3 weeks after initiation of folate/methyl deficiency was accompanied by parallel increases in DNA strand breaks, AP sites and increased levels of AP endonuclease mRNA. In addition, uracil was also detected within the p53 gene sequence using UDG PCR techniques. Increased levels of uracil in DNA implies that the capacity for uracil base excision repair is exceeded with chronic folate/methyl deficiency. It is possible that enzyme induced extrahelical bases, AP sites and DNA strand breaks interact to negatively affect the stability of the DNA helix and stress the structural limits of permissible uracil base excision repair activity. Thus substitution of uracil for thymine induces repair-related premutagenic lesions and a novel form of DNA hypomethylation that may relate to tumor promotion in the methyl-deficient model of hepatocarcinogenesis. PMID- 9395205 TI - Induction of replicative DNA synthesis and PPAR alpha-dependent gene transcription by Wy-14 643 in primary rat hepatocyte and non-parenchymal cell co cultures. AB - Peroxisome proliferators (PP) are known hepatocarcinogens in rats and mice. We have investigated the ability of Wyeth-14 643 (Wy), a PP and potent rodent carcinogen, to induce replicative DNA synthesis and to modulate the levels of peroxisome proliferator activated receptor-alpha (PPAR alpha) transcriptionally dependent genes in primary rat hepatocyte (HPC) cultures and hepatocyte/nonparenchymal cell (HPC/NPC) co-cultures maintained on Matrigel. Four days after plating, cells were treated with Wy and replicative DNA synthesis was quantitated using [3H]thymidine incorporation and specific mRNA transcript levels were determined by reverse-transcriptase polymerase chain reaction (RT-PCR). An increase in HPC replicative DNA synthesis was detected at 48 h in both Wy-treated HPC and HPC/NPC co-cultures relative to controls. This increase was approximately 3- and 6-fold in HPC and HPC/NPC cultures respectively, and was Wy concentration dependent. The levels of PPAR alpha-transcriptionally dependent genes [cytochrome P4504A1, acyl-CoA oxidase (AOxase), and liver-fatty acid binding protein (L FABP)] transcripts were determined as indicators of PPAR alpha activation. These transcripts increased dose-dependently at 48 h in HPC/NPC cultures up to 10 microM Wy. Similarly, RT-PCR product levels were also increased in HPC cultures with 10 microM Wy at 48 h. In conclusion, we have investigated the transcription of PPAR alpha-dependent genes and HPC replicative DNA synthesis by Wy in HPC/NPC co-cultures. Results of this work are clearly more reflective of the known in vivo effects of PP and suggest that HPC/NPC co-cultures are more appropriate than HPC cultures for such studies. The effect of PP on human HPC/NPC co-cultures is currently being investigated in our laboratory in an attempt to assess human risks to these chemicals more directly. PMID- 9395206 TI - Immortalized mouse mammary cells in vivo do not exhibit increased telomerase activity. AB - The acquisition of immortalization is an early and carefully documented event in mouse mammary tumorigenesis. Activation of telomerase activity is one hypothesis to explain the acquisition of immortalization. We examined the activity of telomerase in well-defined immortalized, non-tumor cell populations of mouse mammary tissue in vivo. The results indicated that normal virgin and mid-pregnant mammary gland had low to moderate levels of telomerase activity, respectively. In comparison with the levels detected in pregnant mammary gland, telomerase activity was elevated in mammary tumors in situ and in established mammary cell lines in vitro, both tumorigenic and nontumorigenic; however, hyperplastic alveolar preneoplastic mammary outgrowths and non-tumorigenic ductal outgrowths, both in vivo immortalized populations, had telomerase activity <12% of the levels detected in normal pregnant mammary gland. These results suggest that elevated telomerase activity is not necessary for the immortalization phenotype in in vivo mouse mammary cell populations and that elevated telomerase activity occurs as a late event in mammary tumorigenesis. Furthermore, the data suggest that low levels of telomerase activity are characteristic for mouse mammary epithelial cells and not sufficient for immortalization. These data suggest that other factors play more important roles in the induction and/or maintenance of the immortalization state in mammary cell populations. PMID- 9395207 TI - Inhibition of tumor promoter activity toward mouse fibroblasts and their in vitro transformation by tissue inhibitor of metalloproteinases-1 (TIMP-1). AB - Tissue inhibitor of metalloproteinases-1 (TIMP-1), a natural inhibitor of matrix metalloproteinases (MMPs), is known to inhibit invasion and metastasis of tumor cells. In the present study we examined anti-tumor promoter activity of TIMP-1 and its effect on in vitro cell transformation using BALB/3T3 cells in low serum culture medium. In the dye transfer assay the tumor promoter 12-O tetradecanoylphorbol-13-acetate (TPA) continuously blocked gap-junctional intercellular communication (GJIC) of BALB/3T3 cells in confluent phase. TIMP-1 did not prevent transient inhibition of GJIC induced by TPA, but it quickly restored the reduced GJIC level to the control level. The recovery of GJIC was dependent on the concentration of TIMP-1 from 1 to 1000 ng/ml. In an in vitro two stage transformation assay in which BALB/3T3 cells were treated with 0.5 microg/ml N-metyl-N'-nitro-N-nitrosoguanidine as initiator and 100 ng/ml TPA as promoter, TIMP-1 at concentrations > 10 ng/ml inhibited the focus formation of transformed cells by approximately 60%. TIMP-2 and a synthetic MMP inhibitor showed a similar inhibitory activity on in vitro cell transformation. Furthermore, zymographyic analysis showed that TPA treatment of BALB/3T3 cells induced secretion of gelatinase B and stromelysin-1 into the culture medium. These results indicate that TIMP-1 and TIMP-2 have inhibitory activity on in vitro transformation of cells. It seems likely that TPA-inducible MMPs are involved in carcinogenesis and TIMPs have a protective role against carcinogenesis in vivo. PMID- 9395208 TI - Assessment of the mutagenicity of dichloroacetic acid in lacI transgenic B6C3F1 mouse liver. AB - Dichloroacetic acid (DCA) is a chlorination byproduct found in finished drinking water. When administered in drinking water this chemical has been shown to produce hepatocellular adenomas and carcinomas in B6C3F1 mice over the animal's lifetime. In this study, we investigated whether mutant frequencies were increased in mouse liver using treatment protocols that yielded significant tumor induction. DCA was administered continuously at either 1.0 or 3.5 g/l in drinking water to male transgenic B6C3F1 mice harboring the bacterial lacI gene. Groups of five or six animals were killed at 4, 10 or 60 weeks and livers removed. At both 4 and 10 weeks of treatment, there was no significant difference in mutant frequency between the treated and control animals at either dose level. At 60 weeks, mice treated with 1.0 g/l DCA showed a 1.3-fold increase in mutant frequency over concurrent controls (P = 0.05). Mice treated with 3.5 g/l DCA for 60 weeks had a 2.3-fold increase in mutant frequency over the concurrent controls (P = 0.002). The mutation spectrum recovered from mice treated with 3.5 g/l DCA for 60 weeks contained G:C-->A:T transitions (32.79%) and G:C-->T:A transversions (21.31%). In contrast, G:C-->A:T transitions comprised 53.19% of the recovered mutants among control animals. Although only 19.15% of mutations among the controls were at T:A sites, 32.79% of the mutations from DCA-treated animals were at T:A sites. This is consistent with the previous observation that the proportion of mutations at T:A sites in codon 61 of the H-ras gene was increased in DCA-induced liver tumors in B6C3F1 mice. The present study demonstrates DCA associated mutagenicity in the mouse liver under conditions in which DCA produces hepatic tumors. PMID- 9395209 TI - Dietary energy restriction does not inhibit pancreatic carcinogenesis by N nitrosobis-2-(oxopropyl)amine in the Syrian hamster. AB - Dietary energy restriction was previously shown to be effective in preventing a wide range of experimentally induced cancers. Studies were conducted to assess the influence on pancreatic carcinogenesis of dietary energy restrictions (reduced fat and carbohydrate) of 10%, 20% or 40% in comparison with control in Syrian hamsters treated with N-nitrosobis(2-oxopropyl)amine (BOP). Two carcinogenesis studies were conducted. One used a single treatment with 20 mg BOP/kg body weight and followed hamsters for 102 weeks following treatment, and the other used three weekly treatments of 20 mg BOP/kg body weight and followed hamsters for 45 weeks after treatment. Hamsters were fed control or energy restricted diet beginning the week following the last BOP treatment. Pancreatic carcinomas were induced in 9-18% of the hamsters in the first experiment and in 59-66% of the animals in the second. Dietary energy restriction did not influence carcinoma incidence in either study, and in the second experiment the multiplicity of tumors was higher in the 40% energy restriction (ER) group than in control hamsters. Plasma corticosterone was suppressed by BOP treatment, particularly in the 20% and 40% ER hamsters in the second experiment, and diet or BOP treatment did not significantly alter plasma cortisol. Pancreatic protein kinase Czeta measured by Western blot was highest in the cytosol and particulate fractions of the 40% ER hamsters in the first experiment. These results indicate that dietary energy restriction is not effective in the prevention of BOP induced pancreatic carcinogenesis in the Syrian hamster. PMID- 9395210 TI - Oltipraz stimulates the transcription of the manganese superoxide dismutase gene in rat hepatocytes. AB - Oltipraz (4-methyl-5-(2-pyrazinyl)-1,2-dithiole-3-thione) (OPZ) is recognized as a potent chemoprotective agent against chemical-induced carcinogenesis in several animal models and is thought to act mainly by inducing phase II conjugating together with inhibiting phase I detoxication enzymes. The present study was undertaken to determine whether oltipraz can also influence expression of genes encoding antioxidant enzymes. In rat hepatocytes in primary culture, this compound was found to selectively induce the transcription of the manganese superoxide dismutase (Mn-SOD) gene while it had no effect on copper/zinc-SOD and glutathione peroxidase genes. Oltipraz increased Mn-SOD gene expression in a time and dose-dependent manner by 2- to 3-fold and enhanced the binding activity of the nuclear factor kappa B within 30 min. Moreover, the increase in Mn-SOD gene transcription was associated with a 2- to 3-fold increase of free malondialdehyde and conjugated dienes, two markers of lipid peroxidation, an index of oxidative stress. These results suggest that in rat hepatocytes, oltipraz induced a production of reactive oxygen species that probably acted as second messengers in order to trigger the transcription of many genes. Such a mechanism of action of OPZ and other dithiolethiones would account for the broad spectrum of action of these anticarcinogenic compounds. PMID- 9395211 TI - Prevention by retinoids of azoxymethane-induced tumors and aberrant crypt foci and their modulation of cell proliferation in the colon of rats. AB - Retinoids are proposed chemopreventive agents that inhibit cell proliferation and induce differentiation. Their ability to prevent azoxymethane (AOM)-induced aberrant crypt foci (ACF) and tumors and to modulate cell proliferation was investigated in the colon of male F344 rats. Thirteen retinoids were evaluated for prevention of ACF and two of them, 9-cis-retinoic acid (RA) and 4 (hydroxyphenyl)retinamide (4-HPR), were also evaluated for prevention of colon cancer. The retinoids were administered continuously in the diet starting 1 week prior to the first of two weekly 15 mg/kg i.p. injections of AOM and for a total of either 5 or 36 weeks in order to evaluate their effect on colonic ACF and tumors. At a concentration of 1 mmol/kg diet, 2-(carboxyphenyl)retinamide caused the greatest reduction (57.7%) in the yield of ACF. 9-cis-RA was toxic at 1 mmol/kg so that it was evaluated at 0.1 mmol/kg, resulting in a 41.6% reduction in ACF. The ability of the retinoids to reduce the proliferating cell nuclear antigen (PCNA) labeling index in ACF and in non-involved crypts correlated with their ability to prevent ACF. Both 9-cis-RA (0.1 and 0.2 mmol/kg diet) and 4-HPR (1 and 2 mmol/kg diet) were highly effective in decreasing the yield of AOM induced colon tumors. In summary, retinoids were demonstrated to reduce cell proliferation and to prevent ACF and tumors in the colon, suggesting promise as preventive agents for colon cancer. PMID- 9395212 TI - A prospective study of NAT2 acetylation genotype, cigarette smoking, and risk of breast cancer. AB - Polymorphisms in the N-acetyltransferase 2 (NAT2) gene are determinants of the rate of metabolic activation of carcinogenic compounds such as aryl aromatic amines. Homozygosity for any combination of three variant alleles in Caucasians defines 'slow' acetylators; presence of one or two wild-type alleles characterizes 'rapid' acetylators. Although most previous studies have not observed an overall elevation in risk of breast cancer among slow acetylators, a recent study observed that cigarette smoking was associated with a large increase in risk of breast cancer among slow acetylators. We assessed the relation between NAT2 acetylation status and breast cancer risk, and its interaction with smoking, in a prospective study of mainly Caucasian US women. Four hundred and sixty-six incident cases who were diagnosed with breast cancer after giving a blood specimen in 1989-90 were matched to 466 controls in a nested case-control study. NAT2 genotype was determined using PCR-RFLP assays. The multivariate relative risk (RR) comparing slow with rapid acetylators was 0.9 (95% CI 0.7-1.2). Among slow acetylators, current smoking immediately prior to diagnosis was not associated with a significant elevation in risk compared with never smoking rapid acetylators (RR = 1.4, 95% CI 0.7-2.6). No significant association was seen between pack-years of smoking and risk of breast cancer among either slow or fast acetylators. A non-significant elevation in risk was observed among women who smoked for > or = 5 years prior to first pregnancy and were rapid acetylators, compared with never smoking rapid acetylators (RR = 1.5, 95% CI 0.9-2.6). In analyses limited to 706 post-menopausal women, the elevated risks for current smokers immediately prior to diagnosis who were slow acetylators compared with never smokers who were fast acetylators were slightly stronger but still not statistically significant. In summary, we observed little evidence of an association between NAT2 genotype and breast cancer. In this prospective study, cigarette smoking was not appreciably associated with breast cancer among either slow or fast NAT2 acetylators. PMID- 9395213 TI - Inhibition by N-(4-hydroxyphenyl)retinamide and all-trans-retinoic acid of exogenous and endogenous development of putative preneoplastic, glutathione S transferase placental form-positive lesions in the livers of rats. AB - The effects of N-(4-hydroxyphenyl)retinamide (4-HPR) and all-trans-retinoic acid (tRA) on the exogenous and endogenous models of rat liver carcinogenesis respectively using diethylnitrosamine (DEN) and a choline-deficient, L-amino acid defined (CDAA) diet were studied. For the exogenous study, male Fischer 344 rats, 6 weeks old, were given a single i.p. dose of 200 mg/kg body wt of DEN, partially hepatectomized at week 3, administered 4-HPR at doses of 0, 0.04, 0.08 and 0.16% or tRA at 0, 0.004, 0.008 and 0.015% in diet from week 2 for 6 weeks, and killed at the end of week 8. For the endogenous study, rats were fed the CDAA diet containing 4-HPR or tRA for 12 weeks and killed at the end of week 12. 4-HPR decreased the numbers and sizes of the glutathione S-transferase placental form positive foci, assayed as putative preneoplastic lesions, the levels of 8 hydroxyguanine (8-OHG), a parameter of oxidative DNA damage, and the bromodeoxyuridine labeling indices (BrdU L.I.) by all three doses in the DEN initiated case and, more prominently, in the CDAA diet-associated case. In contrast, while tRA failed to exert inhibitory effects apparently on foci development, 8-OHG formation or BrdU labeling in the DEN-initiated case, it reduced the numbers and sizes of the foci, the 8-OHG levels and the BrdU L.I. by all three doses in the CDAA diet-associated case. Furthermore, both 4-HPR and tRA inhibited the CDAA diet-associated induction of hepatocyte necrosis and connective tissue increase but not intrahepatocellular fat accumulation. These results indicate that 4-HPR exerts chemopreventive effects against the exogenous and endogenous rat liver carcinogenesis, while tRA can inhibit only the latter. PMID- 9395214 TI - Chemopreventive activity of thiol conjugates of isothiocyanates for lung tumorigenesis. AB - A series of L-cysteine (L-Cys), glutathione (GSH), and N-acetyl-L-cysteine (NAC) conjugates of phenethyl (PEITC), benzyl (BITC), and 6-phenylhexyl isothiocyanate (PHITC) were studied for their inhibitory activity toward metabolic activation of the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) in mouse lung microsomes. Selected compounds, PEITC, PEITC-GSH, PEITC-NAC and PHITC-NAC, were also assayed for the potential chemopreventive activity toward NNK-induced lung tumorigenesis in A/J mice. Results showed that PEITC and its conjugates inhibited NNK metabolism with decreasing potency: PEITC < PEITC GSH > PEITC-Cys > PEITC-NAC. PHITC and its GSH and NAC conjugates exhibited nearly 10 times higher inhibitory activity toward NNK metabolism than the PEITC counterparts. In the tumor bioassay, as expected, the conjugates exhibited inhibitory activity against lung tumorigenesis induced by NNK. PEITC-GSH was not inhibitory at 4 micromol/mouse, but it inhibited approximately 32% of lung tumor multiplicity at 8 micromol/mouse. PEITC-NAC at 5 and 20 micromol/mouse both inhibited approximately 30% tumor multiplicity. Among all the conjugates examined, PHITC-NAC was the most potent. At a 5-micromol dose, it completely inhibited tumor multiplicity and incidence to the background level observed in the control group. These results revealed that the structure-activity relationships of the conjugates are similar to those found with their parent isothiocyanates (ITCs), i.e., the potency increased with the increasing alkyl chain length from two to six carbons in arylalkyl ITCs, suggesting that a common active species is involved. The inhibitory activity of ITC conjugates and the expected low toxicity make thiol conjugates of ITC a promising new series of chemopreventive agents. PMID- 9395215 TI - Inhibition of 2-amino-3-methylimidazo[4,5-f]quinoline-DNA adducts by indole-3 carbinol: dose-response studies in the rat colon. AB - Indole-3-carbinol (I3C) inhibits the formation of colonic aberrant crypt foci and DNA adducts in rats given heterocyclic amine colon carcinogens, such as 2-amino-3 methylimidazo[4,5-f]quinoline (IQ). Mechanism studies indicate that I3C induces cytochromes P4501A1 and 1A2 (CYP1A1 and CYP1A2), isozymes that respectively metabolize IQ via ring hydroxylation or activate the carcinogen by N hydroxylation. The present study examined the dose-response for induction of CYP1A1 versus CYP1A2 by I3C, and compared the profiles of induction with the dose response for inhibition of IQ-DNA adducts in the colon of the F344 rat. Dietary equivalent doses of I3C in the range 100-1000 p.p.m. increased in a dose-related manner both ethoxyresorufin O-deethylase (EROD) and methoxyresorufin O demethylase (MROD) activities in the liver and colonic mucosa, and Western blots showed a corresponding induction of CYP1A1 and CYP1A2 proteins. However, dietary equivalent doses of I3C in the range 10-25 p.p.m. (i) reduced hepatic EROD and MROD activities and CYP1A protein levels compared with controls, (ii) increased the ratio of CYP1A2 versus CYP1A1, and (iii) activated IQ to a more potent mutagen when liver microsomes from rats given I3C were used for metabolic activation in the Salmonella assay. Rats given a single oral dose of I3C shortly before administering IQ (5 mg/kg body wt, p.o.) exhibited dose-related inhibition of colonic IQ-DNA adducts in the range 25-100 p.p.m. I3C, reaching 95% inhibition at doses > or = 100 p.p.m. I3C, but IQ-DNA adducts were elevated slightly at the lowest I3C dose as compared with the controls. The possible significance of the low versus high dose effects of I3C are discussed in the context of human dietary exposures to I3C and the reported chemopreventive mechanisms of I3C in vivo. PMID- 9395216 TI - Citrus auraptene inhibits chemically induced colonic aberrant crypt foci in male F344 rats. AB - The modifying effect of dietary administration of auraptene isolated from the peel of citrus fruit (Citrus natsudaidai Hayata) on the development of azoxymethane (AOM)-induced colonic aberrant crypt foci (ACF) was investigated in rats. Male F344 rats were given s.c. injections of AOM (15 mg/kg body wt) once a week for 3 weeks to induce ACF. They also received diets containing 100 or 500 p.p.m. auraptene for 5 weeks, starting 1 week before the first dose of AOM. At termination of the study (week 5) dietary administration of auraptene caused a significant reduction in the frequency of ACF in a dose-dependent manner (P < 0.05). Feeding of auraptene suppressed expression of cell proliferation biomarkers (5-bromo-2'-deoxyuridine labeling-index, ornithine decarboxylase activity, polyamine content and number of silver stained nucleolar organizer region protein particles) in the colonic mucosa and the occurrence of micronuclei caused by AOM. Also, auraptene increased the activities of phase II enzymes (glutathione S-transferase and quinone reductase) in the liver and colon. These findings might suggest that inhibition of AOM-induced ACF may be associated, in part, with increased activity of phase II enzymes in the liver and colon and suppression of cell proliferation in the colonic mucosa. PMID- 9395217 TI - Inhibitory effect of black tea on the growth of established skin tumors in mice: effects on tumor size, apoptosis, mitosis and bromodeoxyuridine incorporation into DNA. AB - Female CD-1 mice were initiated with a single topical application of 7,12 dimethylbenz[a]anthracene and promoted with 12-O-tetradecanoylphorbol-13-acetate. Mice with established papillomas were then treated with black tea or decaffeinated black tea (approximately 4 mg tea solids/ml) as the sole source of drinking fluid for 11-15 weeks. In four separate experiments, oral administration of black tea inhibited the growth of papillomas (increase in tumor volume/mouse) by an average of 35%, 37%, 41% and 48%, respectively. Studies with decaffeinated black tea gave inconsistent results. In one experiment, administration of decaffeinated black tea inhibited papilloma growth (increase in tumor volume/mouse) by 27%, but in two additional experiments papilloma growth was stimulated by 14% and 193%, respectively. In a separate experiment, skin tumors were generated by treating SKH-1 female mice with ultraviolet B light (UVB; 30 mJ/cm2) twice weekly for 22 weeks, after which UVB administration was stopped. Tumors were allowed to develop during the following 13 weeks, and tumor-bearing mice were then treated with black tea (6 mg/ml tea solids) as the drinking fluid for 11 weeks. In this experiment, tumor growth (increase in tumor volume/mouse) was inhibited by 70%. Histological examination revealed that tea-treated mice had a 58% decrease in the number of nonmalignant tumors (primarily keratoacanthomas)/mouse and a 54% decrease in the number of squamous cell carcinomas/mouse. In addition, administration of black tea decreased the volume per tumor by 60% for nonmalignant tumors and by 84% for carcinomas. Mechanistic studies with tumors from these mice revealed that administration of black tea decreased the bromodeoxyuridine labeling index in squamous cell papillomas, keratoacanthomas and squamous cell carcinomas by 56%, 45% and 35%, respectively, and the apoptosis index was increased by 44%, 100% and 95%, respectively. Administration of black tea decreased the mitotic index in keratoacanthomas and squamous cell carcinomas by 42% and 16%, respectively. The results indicate that oral administration of black tea to tumor-bearing mice inhibited proliferation and enhanced apoptosis in nonmalignant and malignant skin tumors. PMID- 9395218 TI - Metabolic competence and susceptibility of intestinal epithelium to genotoxic injury during regeneration. AB - The carcinogenic potency of many mutagens is increased in conditions of tissue regeneration. This involves fundamental changes of cellular division and differentiation, in intestinal epithelium. However, effects on epithelial capacity for carcinogen metabolism and susceptibility to genotoxic injury are unknown. Using a novel rat model, this study assessed expression of cytochrome P450 mono-oxygenases (Cyps), glutathione S-transferases (GSTs) and uridine diphosphoglucuronosyl transferase (UGT) in intestinal epithelium during sequential stages of regeneration. Enzyme induction and DNA adduct formation were also assessed after benzo[a]pyrene (BaP) exposure. Control assays were carried out in normal intestinal epithelium. Fewer phase I and II xenobiotic metabolizing enzymes were expressed in regenerating intestinal epithelium than in normal control intestinal epithelium (GSTA3, UGT in regeneration vs Cyp2B, GSTA1/2, GSTA4, GSTP1, UGT in control). Benzo[a]pyrene induced GSTA3 and UGT in regeneration vs Cyp1A, Cyp2B, GSTA1/2, GSTA3, GSTA4, GSTP1 and UGT in control normal intestinal epithelium. Benzo[a]pyrene induced low levels of GSTA3 in early regenerating intestinal epithelium but induction increased by >2-fold at late stage regeneration. Higher levels of benzo[a]pyrene 7,8-diol-9,10-epoxide (BPDE) DNA adducts were formed at early stages of regeneration, than at later stages. Intestinal epithelium displayed reduced metabolic competence and differential susceptibility to genotoxic injury from BaP, during regeneration. PMID- 9395220 TI - DNA damage and mutagenesis induced by procarbazine in lambda lacZ transgenic mice: evidence that bone marrow mutations do not arise primarily through miscoding by O6-methylguanine. AB - The DNA damaging and mutagenic activities of procarbazine, a methylating drug employed in cancer chemotherapy and suspected of causing therapy-related leukaemia, were investigated in the liver and bone marrow of lambda lacZ transgenic mice (MutaMouse). The drug was administered using two different protocols, a 'high-dose' one involving 5 daily doses of 200 mg/kg, expected to cause depletion of the repair enzyme O6-alkylguanine-DNA alkyltransferase (AGT) and thus favour the selective accumulation of the premutagenic lesion O6 methylguanine (O6-meG) relative to other adducts, and a 'low-dose' one involving 10 daily doses of 20 mg/kg procarbazine. Substantial accumulation of O6-meG was observed in both tissues examined 6 h after the end of the 'high-dose' treatment, with the liver accumulating somewhat higher levels than the bone marrow (28.0 +/- 1.8 fmol/microg DNA and 18.5 +/- 1.1 fmol/microg DNA respectively). However, significant increases in mutant frequency 10 days after the end of treatment were observed only in the bone marrow, reaching a 16-fold increase over background following the 5 x 200 mg/kg treatment. Sequence analysis of the mutations induced after this treatment revealed a mixed spectrum, in which G:C-->A:T transitions (characteristic of O6-meG miscoding) were only a secondary feature: Among 20 mutants analysed, only six such mutations were found, including three at CpG sites, which might have arisen from deamination of 5-methylcytosine. The other mutations observed included 1 A:T-->G:C transition, five transversions (one G:C- >T:A, one double G:C-->C:G, two A:T-->T:A, one A:T-->C:G), five deletions and three insertions. The mechanistic and clinical significance of these findings is discussed. PMID- 9395219 TI - Distinct time courses of increase in cytochromes P450 1A2, 2A5 and glutathione S transferases during the progressive hepatitis associated with Helicobacter hepaticus. AB - Mice naturally infected by Helicobacter hepaticus develop a chronic active hepatitis leading to hepatocellular carcinoma. This mouse model of liver cancer was used to examine the impact of bacterial infection on the hepatic expression and activity of enzymes involved in carcinogen bioactivation (phase I enzymes) and detoxification (phase II enzymes). No major differences in total cytochrome P450 (CYP) content were found between control and infected mice during the course of the study. The most striking modulations of individual isoenzymes were the increases in immunohistochemical staining observed for CYP1A and CYP2A5 in relation to increasing age and liver lesions. The increase in CYP2A5 in mice aged over 12 months was confirmed by the observed increases in coumarin 7 hydroxylation (CYP2A5 substrate) in vitro and CYP2A5 mRNA levels by Northern blot analysis. Immunoblotting confirmed the specific induction of CYP1A2 in infected mice 12 and 18 months of age. Perfusion of liver with nitroblue tetrazolium, an indicator for superoxide formation, demonstrated that in livers of infected mice, hepatocytes often co-expressed CYP2A5 and formazan deposition. Concerning phase II enzymes, an enhancement of glutathione S-transferase (GST) activities, related to the disease process, was observed in infected mice. An age-specific increase of GSTpi and A4.4 (early stage of disease) and GST YaYa (>9 months) expression was also demonstrated by immunohistochemical staining. In contrast, catalase and glutathione-peroxidase activities, as well as reduced glutathione content were decreased in the early stages of disease (3-9 months) in infected mice compared to age-matched control mice. Overall, these results suggest that alterations in CYP and GST expression may contribute to the aetiology of tumour incidence due to H. hepaticus infection via production of reactive oxygen species. PMID- 9395221 TI - Intestinal and extra-intestinal tumor multiplicities in the Apc1638N mouse model after exposure to X-rays. AB - Seven-week-old Apc1638N mice were exposed to a single dose of 5 Gy total-body X irradiation resulting in a 8-fold increase in the number of intestinal tumors and a reduction of the lifespan to an average of 6 months. The distribution of tumors along the intestinal tract as well as the adenoma/carcinoma ratio, were similar between non-irradiated and irradiated animals. Semi-quantitative PCR analysis of intestinal-tumor DNA revealed that 10 out of 14 tumors had lost the wild-type Apc allele. However, in contrast to spontaneous Apc1638N intestinal tumors in which the LOH event at the Apc locus involves the entire chromosome 18 (1), in 6 out of 10 tumors derived from X-irradiated animals the Apc loss is associated with only a partial intrachromosomal deletion. The remaining tumors have lost all chromosome 18 markers tested. In addition to the intestinal tumors, female Apc1638N mice are susceptible to the development of mammary tumors. Upon X irradiation, Apc1638N mice show a striking 15-fold increase in mammary tumors. Moreover, Apc1638N mice spontaneously develop other extra-intestinal neoplasia, such as desmoid-like lesions similar to those associated with familial adenomatous polyposis (FAP), the human syndrome caused by germline mutations in the APC gene. Spontaneous desmoid growth is sex-dependent, as male Apc1638N mice develop 3-fold more desmoids than female mice. Interestingly, X-irradiation seemed to increase the number of desmoids per animal nearly twofold only in female Apc1638N mice. Five out of 9 desmoids found in Apc1638N mice exposed to X ray displayed loss of the wild-type Apc allele. PMID- 9395222 TI - DNA adducts in human nasal mucosa and white blood cells from smokers and non smokers. AB - The goal of the present study was to measure the levels of DNA adducts in human nasal mucosa cells and in total white blood cells in relation to smoking. DNA was isolated from samples of 20 healthy volunteers (six smokers and 14 non-smokers). The levels of DNA adducts were measured by 32P-postlabelling assay. In smokers the mean DNA adduct levels were 3.3 and 17.0 adducts/10(8) nucleotides in total white blood cells and nasal mucosa cells respectively. The corresponding values in non-smokers were 2.0 and 6.8 adducts/10(8) nucleotides. The mean adduct level was significantly higher in nasal mucosa cells than in total white blood cells both in smokers and non-smokers. The mean adduct levels in smokers' nasal mucosa cells were significantly higher than those in non-smokers. Thus the nasal mucosa cells constituted a sensitive tissue for the determination of cigarette smoking induced DNA adducts. Combining the sensitivity of the 32P-postlabelling assay with the specificity of the nasal mucosa to the airborne chemical exposures, the DNA adduct analysis from human nasal mucosa cells represents a method of choice in the assessment of exposure to airborne carcinogens. PMID- 9395223 TI - Investigation of the formation and accumulation of liver DNA adducts in mice chronically exposed to tamoxifen. AB - Tamoxifen was administered to three strains of female mice (B6C3F1, C57BL/6 and DBA/2) in short- and long-term studies to determine their ability to activate tamoxifen and cause hepatic DNA damage. 32P-Postlabelling of liver DNA from mice treated for 4 days showed a group of major adducts that increased in a dose dependent manner and co-chromatographed with the major adducts detected in rat liver. On cessation of dosing, the majority of adducts were cleared within 3 days. Binding of [14C]tamoxifen to DNA nucleotides was demonstrated by the use of accelerator mass spectrometry. In long-term studies of 12 months to 2 years duration, dependent on strain, tamoxifen was administered continuously in the diet to give a daily dose of approximately 40 mg/kg. DNA adducts were detected after 3 months, although the number of adducts decreased with time and by 2 years were not detectable in the tamoxifen treated mice. None of the treated groups showed a significantly increased incidence of liver tumours, with or without phenobarbital promotion and there was no sustained liver cell proliferation. Tamoxifen was detected in the mouse livers, but at levels 50 times lower than those reported in a comparable rat study. These results suggest that, in contrast to the rat, tamoxifen is non-carcinogenic in mice because it does not cause sufficient cumulative DNA damage, or act as a promoter by causing cell proliferation. PMID- 9395224 TI - Dysregulation of ornithine decarboxylase activity, apoptosis and Bcl-2 oncoprotein in Syrian hamster embryo cells stage-exposed to di(2 ethylhexyl)phthalate and tetradecanoylphorbol acetate. AB - Perturbations of cell proliferation and death are considered as essential events in the process of carcinogenesis. Thus, two parameters, ornithine decarboxylase (ODC), an enzyme closely related to cell proliferation and transformation, and apoptotic phenomenon are profoundly modified. Using Syrian hamster embryo (SHE) cells, we have examined in the framework of two-stage carcinogenesis (initiation promotion) the effects of a non-genotoxic [diethylhexylphthalate (DEHP)] or genotoxic [benzo[a]pyrene (BaP)] carcinogen or a non-carcinogenic compound [phthalic anhydride (AP)] on these parameters. Immunoreactive Bcl-2 and Bcl-xL proteins were also investigated following two-stage exposures. Whereas exposures to BaP, DEHP or AP alone did not provoke any modification of ODC activity, the phorbol ester, 12-O-tetradecanoylphorbol-13-acetate (TPA), strongly increased it. Using two-stage exposure protocol (xenobiotics first, then replacement by TPA promoter), the ODC activity was higher than that obtained with TPA alone. This superinduction of ODC activity was observed only with the carcinogenic compounds DEHP and BaP. Following the same exposure protocol, spontaneous cellular apoptosis was decreased. Furthermore, Bcl-2 oncoprotein was also upregulated approximately 8- and 11-fold for BaP and DEHP respectively; meanwhile Bcl-xL protein rate did not change. The non-carcinogenic compound AP slightly inhibited spontaneous SHE cell death without ODC superinduction. Exogenous polyamines, putrescine, spermidine and spermine diluted in the medium did not inhibit spontaneous apoptosis. Although inhibition of apoptosis was not specific of carcinogenic compound, both superinduction of ODC activity and inhibition of apoptosis via Bcl-2 upregulation, may cooperate during early stages of the carcinogenic process. PMID- 9395225 TI - Determination of steady-state levels of oxidative DNA base modifications in mammalian cells by means of repair endonucleases. AB - The alkaline elution technique in combination with various repair endonucleases (Fpg protein, endonuclease III, exonuclease III, T4 endonuclease V) was used to quantify steady-state (background) levels of oxidative base modifications in various types of mammalian cells. In human lymphocytes the number of base modifications sensitive to Fpg protein, which include 8-hydroxyguanine, was 0.25 +/- 0.05 per 10(6) base pairs. Even lower levels (0.07 +/- 0.02 per 10(6) bp) were observed in HeLa cells. The numbers of sites sensitive to the other repair endonucleases were below the detection limit (0.05 per 10(6) bp). In a direct comparison, the background level of Fpg-sensitive modifications determined by alkaline elution was much lower than the background level of 8 hydroxydesoxyguanosine (8-oxodG) determined after enzymatic DNA hydrolysis by HPLC and electrochemical detection. However, the number of additional Fpg sensitive modifications induced by a photosensitizer plus light was similar to the additional number of 8-oxodG residues determined by HPLC with electrochemical detection. This indicates that the enzyme assay does not systematically underestimate the number of lesions and points to an artefactual generation of 8 oxodG during DNA isolation and hydrolysis. PMID- 9395226 TI - The mutagenic response at the ouabain resistance locus in T cells of mice exposed to N-ethyl-N-nitrosourea parallels the response at the Hprt locus and correlates with mutation target size. AB - The lymphocyte Hprt gene has been used extensively as a reporter locus to monitor the mutational effects of the exposure of animals to genotoxicants. Implicit in this view of the function of a reporter gene is the assumption that its mutagenic response is representative of that of other genes in the organism. As a test of this hypothesis we compared the frequency of 6-thioguanine-resistant (TGr) mutants at the Hprt locus with the mutant frequency (MF) induced at another locus, the ouabain resistance (Oua) locus. The frequency of spontaneous OUA(R) mutants was estimated to be 1.1x10(-7) (MF between <0.3 and 1.1x10(-7)), which was approximately 30-fold less than the spontaneous TGr MF. Following treatment with N-ethyl-N-nitrosourea (ENU), the induced OUA(R) MF at each of two dose levels (50 and 150 mg/kg ENU) and two time points (3 and 6 weeks post-exposure) was consistently 8- to 9-fold lower than the corresponding TGr MF. Thus the mutagenic response of the Oua locus closely paralleled that of the Hprt locus, indicating a similarity in their response to ENU. In addition, the Oua locus was 3-4 times more sensitive than the Hprt locus to the mutagenic effect of ENU, as measured by the fold increase in MF over the background level. The number of ENU mutable sites capable of resulting in a TGr or OUA(R) phenotype, otherwise known as the mutation target size, was estimated to differ by an order of magnitude between the two loci. This difference in target size correlates with, and therefore may largely account for, the difference in induced MF between both loci. PMID- 9395227 TI - Analysis of tissue-specific lacZ mutations induced by N-nitrosodibenzylamine in transgenic mice. AB - N-Nitrosodibenzylamine (NDBzA) is a contaminant found frequently in rubber baby bottle nipples and pacifiers. To evaluate more fully the mutagenic potential and analyse the molecular nature of possible mutations induced in vivo, we have studied the mutagenicity of NDBzA in vivo using the MutaMouse system. NDBzA, suspended in olive oil, was administered orally once to male mice at different doses (0, 30, 100, 425 and 750 mg/kg) and the mice were killed 30 and 90 days after treatment. As a positive control, and to compare relative mutagenicity, N nitrosodimethylamine (NDMA) was also administered to animals in the same experiment at doses of 0, 2, 6 and 10 mg/kg. Mutant frequencies were increased in both 30 and 90 day liver samples, but not in bone marrow, after both NDBzA and NDMA treatment. However, NDBzA was >100 times less mutagenic than NDMA. A total of 81 mutants obtained from liver samples of treated animals (750 mg/kg) were characterized by DNA sequencing. While spontaneous mutations in transgenic mice have been characterized previously by a preponderance of G:C-->A:T transitions, mainly at 5'-CpG-3' dinucleotide sites, the predominant type of NDBzA-induced mutation in this study was transversion, mainly G:C-->T:A changes. The molecular characteristics of mutations induced by NDBzA indicate that they may arise from specific unidentified DNA adducts and benzylation appears to be the primary mechanism involved in formation of these DNA adducts. PMID- 9395228 TI - Initiating activity of the anti-estrogen tamoxifen, but not toremifene in rat liver. AB - A striking difference between two structurally related anti-estrogen medicines is that tamoxifen is strongly hepatocarcinogenic in the rat, whereas toremifene lacks such activity. To study the basis for this difference, the initiating potential of tamoxifen and toremifene were studied by measurement of rapid induction of hepatocellular altered foci (HAF) that express placental-type glutathione S-transferase in the livers of female Sprague-Dawley (S-D) rats and female Fischer 344 (F344) rats. Both agents were administered by gavage at equimolar doses up to a dose that produced marked weight gain suppression. In rats given the high dose of 40 mg/kg per day tamoxifen continuously for 36 weeks, 75% of S-D rats developed liver neoplasms, in contrast to only 10% of F344 rats. In the S-D strain, tamoxifen produced a tendency to increased HAF at 2 weeks at the dose of 40 mg/kg per day and by 12 weeks, a dose-related increase was evident. In contrast, toremifene induced no HAF even at the equimolar high dose of 42.4 mg/kg per day for 12 weeks. The induction of HAF by tamoxifen was less in the F344 rats. Neither agent elicited increases in hepatocellular proliferation in S-D or F344 rats. When phenobarbital was administered for 24 weeks as a promoting agent after the anti-estrogens, S-D rats given tamoxifen at 20 mg/kg per day for 12 weeks, developed liver neoplasms, but not F344 rats or rats of either strain given even a higher dose (42.4 mg/kg) of toremifene. Thus, tamoxifen has initiating activity in these rat strains whereas toremifene does not. PMID- 9395229 TI - Functional characterization of the rat mdr1b encoded P-glycoprotein: not all inducing agents are substrates. AB - The multidrug resistance (mdr) genes encode P-glycoproteins, integral membrane proteins which function as drug efflux transporters. Exposure of animals in vivo and cells in vitro to a variety of xenobiotics leads to increased mdr1 gene expression and higher levels of P-glycoprotein. This response may protect cells from the cytotoxic effects of these compounds. In this investigation we functionally expressed the rat mdr1b gene in NIH 3T3 cells and assessed the ability of the encoded P-glycoprotein to protect these cells from the cytotoxicity of xenobiotics known to induce mdr1b expression. In long-term colony survival assays, stably expressed mdr1b conferred resistance to cytotoxic drugs such as colchicine, vinblastine and doxorubicin, but not to 5-fluorouracil nor to the carcinogens aflatoxin B1 and N-hydroxy-acetylaminofluorene. The mdr reversal agent verapamil restored cytotoxicity of colchicine, doxorubicin, actinomycin D, vinblastine and taxol, but had no effect on the sensitivity of these cells to 5 fluorouracil, aflatoxin B1 or N-hydroxy-acetylaminofluorene. In a competitive transport assay, verapamil and, to a lesser extent, colchicine blocked the increased efflux of the fluorescent dye rhodamine 123 from mdr1b-transfected cells, whereas aflatoxin B1 did not compete for this export. These data demonstrate that expression of the rat mdr1b encoded P-glycoprotein can protect cells from a diverse group of compounds previously identified to be mdr substrates, however, other effective inducers of mdr expression, such as aflatoxin B1 and N-hydroxy-acetylaminofluorene, remain potent cytotoxins despite high levels of P-glycoprotein. The fact that compounds which are not themselves substrates can induce P-glycoprotein expression may have implications for pharmacokinetic interactions and chemotherapy. PMID- 9395230 TI - Development of esophageal metaplasia and adenocarcinoma in a rat surgical model without the use of a carcinogen. AB - In order to establish an animal model for studying the cause and prevention of esophageal adenocarcinoma (EAC) and its frequent precursor, Barrett's esophagus (BE), factors affecting the pathogenic processes were investigated in an esophagoduodenal anastomosis model with rats. Experiments by us and others have shown that surgical treatment produced reflux esophagitis with cell hyperproliferation, but not EAC. Additional treatment with a carcinogen has been shown to be necessary for the development of EAC, squamous cell carcinomas (SCC) or EAC/SCC mixtures. We found that the surgically treated animals developed anemia due possibly to reduced iron absorption. When the operated animals were supplemented with iron, EAC occurred at a high rate (73%) after 30 weeks, and treatment with N'-nitrosonornicotine did not enhance the rate of tumorigenesis. Treatment with carcinogen, however, induced SCC in the group of rats killed after 22 weeks. The results suggest that iron overload, which is known to cause oxidative damage, is an enhancing factor for adenocarcinogenesis. The pathogenesis of EAC in the iron-supplemented, non-carcinogen treated group resembles human esophageal adenocarcinogenesis in many features. All the BE was the specialized type with goblet cells (containing sialomucin or sulfomucin) and columnar cells (containing acid or neutral mucin) as well as an incompletely developed brush border. Almost all of the BE was located at the bottom of the esophagus and was continuous with the duodenal mucosa; dysplasia became more frequent at later time points. All of the cancers were well-differentiated mucinous EAC, and most of the EAC had an adjacent area of BE with dysplasia. The results are consistent with the proposed human sequence for pathogenic events of BE progression to 'BE with dysplasia' and then to EAC. Esophagoduodenal anastomosis and iron treatment in rats produces a high rate of BE and EAC which are morphologically similar to human BE and EAC; this may be a useful animal model to study the development and prevention of EAC in humans. PMID- 9395231 TI - Cyclic food restriction, insulin and mammary cell proliferation in the rat. AB - We reported recently that weight cycling significantly increased the incidence of mammary cancer in virgin female rats that were pretreated with N-methyl-N nitrosourea. The present study investigated the effect of weight cycling on mammary epithelial cell proliferation and its relationship to changes in plasma insulin, estrogen, progesterone and urinary corticosterone in 30 female virgin Sprague-Dawley rats. Animals were fed a modified AIN-76A diet containing 24.6% corn oil by weight. Weight-cycled (WC) rats were food restricted daily by either 33% or 50% of non-restricted controls for 1 week followed by 3 weeks compensatory refeeding and weight recovery over 18 weeks or 4.5 weight cycles. WC rats consumed 6-10% less food than controls (P = 0.01) but showed a 71-89% greater efficiency of food utilization for growth (P < 0.0001) than controls. There were no differences in total weight gain during treatment. Mammary lobuloalveolar and ductal cell proliferation of WC rats, measured by 5-bromo-2'deoxyuridine labelling, increased in a dose-response fashion, P = 0.03, P = 0.06 respectively in comparison to controls. Energy and substrate utilization measured by indirect calorimetry indicated WC animals expended less energy (P = 0.005) and utilized less glucose (P = 0.0001) and protein (P = 0.006) during restriction, and less lipid during recovery (P = 0.05) than controls. There were no significant differences in hormone levels between groups. Multiple regression analysis with plasma insulin, estrogen, progesterone and urinary corticosterone as independent variables (r = 0.947, r2 = 0.897, P = 0.003) showed that plasma insulin was the only significant predictor (P < 0.01) of mammary cell proliferation. In accord with this observation, tyrosine-phosphorylated activation of insulin receptor substrate-1, detected by immunoprecipitation and Western immunoblot analysis in mammary tumors of WC rats from our previous study, was 3-5 times greater than in non-restricted controls (P < 0.01). Present findings suggest that weight cycling in rats increases risk of breast cancer development via insulin stimulated mammary cell proliferation. PMID- 9395232 TI - Tumour necrosis factor alpha (TNF alpha) suppresses apoptosis and induces DNA synthesis in rodent hepatocytes: a mediator of the hepatocarcinogenicity of peroxisome proliferators? AB - Peroxisome proliferators (PPs) are a class of non-genotoxic rodent hepatocarcinogens that cause increased hepatocyte DNA synthesis, peroxisome proliferation and liver enlargement. We have demonstrated previously that PPs suppress both spontaneous rat hepatocyte apoptosis and that induced by the physiological negative regulator of liver growth, transforming growth factor beta (TGF beta1). Evidence suggests that the suppression of apoptosis by PPs is mediated via activation of the peroxisome proliferator activated receptor-alpha (PPAR alpha), a member of the nuclear hormone receptor superfamily. Here, we investigate the effects of tumour necrosis factor alpha (TNF alpha) on cultured rat or mouse hepatocytes to determine whether TNF alpha influences hepatocyte growth in a manner analogous to that seen with PPs. Rat recombinant TNF alpha was found to stimulate DNA synthesis and suppress apoptosis in isolated rat hepatocyte monolayers (P < or = 0.01). These effects were seen in the range of 500-5000 U/ml with a maximum effect at 5000 U/ml. Similarly, mouse recombinant TNF alpha was able to stimulate DNA synthesis in mouse hepatocyte monolayers (P < or = 0.01) with a maximal effect at 1000 U/ml. Suppression of mouse hepatocyte apoptosis by TNF alpha was not detected, possibly because of the low levels of apoptosis under control conditions. However, when the levels of mouse hepatocyte apoptosis were augmented using TGF beta1, TNF alpha caused a significant suppression (P < or = 0.01). The neutralization of TNF alpha using anti-TNF alpha antibodies abrogated significantly (P < or = 0.01) the suppression of apoptosis by the PP, nafenopin. These data that suggest TNF alpha may mediate, at least in part, the growth perturbation, liver enlargement and hepatocarcinogenesis seen in response to the PP class of non-genotoxic hepatocarcinogens. PMID- 9395233 TI - Artificial background and induced levels of oxidative base damage in DNA from human cells. AB - The pre-mutagenic oxidative DNA base damage of 8-hydroxy-guanine is present in DNA isolated from cells and the amount present increases with exposure of cells to oxidative stress. The oxidative DNA base damage may be present before isolation of DNA or it may be produced during isolation and processing of DNA. We have found that the amount of oxidative base damage measured in DNA can be reduced to a stable lower level by adding increasing concentrations of the antioxidants desferrioxamine, histidine and reduced glutathione immediately before cell lysis. Inclusion of these antioxidants after cell lysis did not affect the level of DNA damage. Oxidative DNA base damage produced by ultraviolet A irradiation of human cells was also reduced by adding antioxidants after irradiation and before cell lysis. Thus, unidentified oxidants induced by ultraviolet A irradiation may damage DNA significantly during extractions of DNA from cells subsequent to ultraviolet A irradiation. PMID- 9395234 TI - Mutation and downregulation of the transforming growth factor beta type II receptor gene in primary squamous cell carcinomas of the head and neck. AB - In the present study, we analyzed 28 squamous cell carcinomas of the head and neck (SCCHN) for mutations in the coding region of TbetaR-II using 'Cold' SSCP and automatic DNA sequencing analyses. Twenty-one percent (6/28) of the SCCHN examined contained TbetaR-II mutations compared with patient-matched normal tissues. These alterations included five missense mutations (A:T-->G:C transitions in codons 250, 401, 448 and 488, and a G:C-->T:A transversion in codon 373), and a 38-bp deletion between nucleotides 1825 to 1862. In addition to these code-altering mutations, one case exhibited a silent mutation (A:T-->G:C transition in codon 451) and three cases contained one of two potential population polymorphisms (codons 354 and 389). In contrast to colon and gastric cancers exhibiting microsatellite instability (MI) or replication errors (RER+), no 'indirect' frameshift mutations were identified within a 10-bp polyadenine repeat present in the TbetaR-II coding sequence. All of the mutations in the present study occurred within the highly conserved serine/threonine kinase domain and represent the first report of such 'direct' TbetaR-II mutations in primary human tumors. In addition, we analyzed a subset of SCCHN and corresponding normal samples for TbetaR-II mRNA expression using semi-quantitative multiplex RT-PCR. Expression of TbetaR-II was decreased by 24% to 74% in 20 of 23 SCCHN (87%) compared with patient-matched normal tissues. Taken together, the results from this study suggest that alterations in the nucleic acid sequence and mRNA expression of TbetaR-II are prevalent events in the development of SCCHN, which may deregulate cell cycle control. PMID- 9395235 TI - Signaling through the ARK tyrosine kinase receptor protects from apoptosis in the absence of growth stimulation. AB - ARK (AXL) is the prototype of a distinctive family of receptor tyrosine kinases which contain in their extracellular domains features reminiscent of cell adhesion molecules. ARK is capable of homophilic binding, which results in a degree of receptor activation, but can also be activated by a heterophilic ligand, Gas6, a member of the family of vitamin K dependent proteins that is preferentially expressed in quiescent cells. Since a number of tissues and cell lines express both ARK and Gas6, we studied the effect of endogenous and exogenous Gas6 on the phenotype of ARK expressing cells. Here we show that constitutive expression of Gas6 in an NIH3T3 cell line that does not spontaneously express this protein does not result in cell transformation or uncontrolled growth, but protects from apoptosis induced by serum deprivation. Recombinant exogenous Gas6 was also capable of protecting cells from apoptosis at concentrations that did not result in significant induction of DNA synthesis. Activation of ARK phosphorylation and a weak but significant induction of MAP kinase activity accompanied the increased survival of cells treated with Gas6. The antiapoptotic effect of ARK signaling was confirmed by studies using fibroblasts from ARK knock-out mice, that showed that the absence of ARK resulted in higher levels of serum deprivation-induced apoptosis, that could not be rescued by the addition of Gas6. Interestingly ARK signaling protects from apoptosis induced by serum deprivation, myc overexpression, or by TNF alpha but not from u.v. irradiation or Staurosporine. These results suggest that a major function of Gas6-ARK signaling is that of increasing cell survival under conditions which do not allow cell proliferation. PMID- 9395236 TI - Different HPV16 E6/E7 oncogene expression patterns in epithelia reconstructed from HPV16-immortalized human endocervical cells and genital keratinocytes. AB - Human papillomavirus type 16 (HPV16) E6/E7 oncogenes immortalize two types of human genital epithelial cells in vitro, endocervical cells and ectocervical or foreskin keratinocytes. Epithelia reconstructed in in vivo nude mouse implants or in vitro organotypic raft cultures from immortalized endocervical cells form higher grade dysplasia than those from keratinocytes. Here, we compared viral E6/E7 mRNA expression in immortalized cell lines of the three cell types using implants, rafts and in situ hybridization assays. Endocervical cells expressed E6/E7 throughout their reconstructed epithelia. In contrast, oncogenes were limited to basal cells for keratinocyte lower grade dysplasias. To study the role of the HPV16 promoter/enhancer in this repression in the upper layers of keratinocyte epithelia, new cell lines were established by immortalization with E6/E7 controlled by the SV40 promoter. The oncogenes were shown to be controlled from the SV40 elements after immortalization. Nevertheless, E6/E7 in the two cell types had the same cell-specific expression pattern as that controlled from the homologous HPV16 promoter. In addition, naturally occurring premalignant lesions having integrated HPV16 DNA expressed E6/E7 extensively in the high-grade dysplastic region of undifferentiated metaplasia. On the other hand, oncogene expression was restricted to lower layers in the lower grade dysplastic region of more mature differentiation. Our data suggest that keratinocytes have an inherent HPV16 promoter-nonspecific mechanism of repression. Apparently this mechanism, which can be acquired during maturation, is initially nonfunctional in in vitro and in vivo epithelia derived from metaplastic endocervical cells. PMID- 9395237 TI - Novel small GTPase M-Ras participates in reorganization of actin cytoskeleton. AB - During an attempt to elucidate regulatory mechanisms of skeletal muscle cell differentiation, we cloned cDNAs encoding a novel small GTPase from cDNA libraries of the mouse skeletal muscle cell line C2 and rat brain. It was designated as M-Ras due to the structural similarity to Ras family proteins. M Ras contained conserved motifs for GDP/GTP-binding and GTPase activities, whereas it varied from the other Ras family proteins at several amino acids within the extended effector domain. From the C-terminal sequence, M-Ras is presumed to be anchored to the cell membrane with a geranylgeranyl group in combination with a polybasic region. Bacterially expressed recombinant M-Ras exerted the GTP-binding and GTPase activities. A mutant M-RasG22V was unable to hydrolyze bound GTP, indicating that it serves as a constitutively active form. Epitope-tagging experiments showed that M-Ras was concentrated on certain plasma membrane associated structures. Transfection of M-Ras cDNA and microinjection of the M RasG22V protein into fibroblasts induced formation of the peripheral microspikes. In addition, the actin stress fibers disappeared and instead numerous actin foci were formed in the injected cells. The transfected cells eventually exhibited dendritic appearances with microspikes. Consequently, M-Ras is likely to participate in reorganization of the actin cytoskeleton. PMID- 9395238 TI - Transcription of the SCL gene in erythroid and CD34 positive primitive myeloid cells is controlled by a complex network of lineage-restricted chromatin dependent and chromatin-independent regulatory elements. AB - The SCL gene (also known as TAL-1) encodes a basic helix-loop-helix transcription factor that is essential for the development of all haematopoietic lineages, and ectopic expression of which results in T cell leukaemia. SCL is expressed in normal pluripotent haematopoietic stem cells and its expression is maintained during differentiation along erythroid, mast and megakaryocytic lineages, but is extinguished following commitment to other cell types. The mechanisms responsible for this pattern of expression are poorly understood, but are likely to illuminate the molecular basis for stem cell development and lineage commitment. We have identified multiple lineage-restricted DNase I hypersensitive sites in a 45 kb region spanning the murine SCL locus. Committed erythroid cells and CD34 positive primitive myeloid cells exhibited both shared and unique DNase I hypersensitive sites whereas none were found in T cells. The function of each hypersensitive site was studied using both transient and stable reporter assays in erythroid, primitive myeloid and T cells. Multiple positive and negative regulatory elements were characterised and found to display lineage-specificity, promoter-specificity and/or chromatin-dependence. These results represent the first description of key components of a complex network of regulatory elements controlling SCL expression during haematopoiesis. PMID- 9395240 TI - Potentiation of apoptosis by low dose stress stimuli in cells expressing activated MEK kinase 1. AB - MEK kinases (MEKKs) are serine-threonine kinases that regulate sequential protein phosphorylation pathways involving mitogen-activated protein kinases (MAPKs), including members of the Jun kinase (JNK) family. MEKK1 is a 196 kDa protein that when cleaved by caspase-3-like proteases generates an active COOH-terminal kinase domain. Expression of the MEKK1 kinase domain is sufficient to induce apoptosis. Mutation of MEKK1 to prevent its proteolytic cleavage protects cells from MEKK1 mediated cell death even though the JNK pathway is still activated, indicating that JNK activation is not sufficient to induce cell death. The inducible acute expression at modest levels of the activated MEKK1 kinase domain can be used to potentiate the apoptotic response to low dose ultraviolet irradiation and cisplatin. Similarly, in L929 fibrosarcoma cells inducible acute expression of the kinase domain of MEKK1 markedly increased the cell death response to tumor necrosis factor alpha (TNF alpha). The findings demonstrate that acute expression of an active form of MEKK1 can potentiate the cell death response to external stress stimuli. Manipulation of MEKK1 proteolysis and its regulation of signal pathways involved in apoptosis has significant potential for anticancer therapies when used in combination with therapeutic agents at doses that alone have little or modest effects on cell viability. PMID- 9395239 TI - p53-induced apoptosis in the human T-ALL cell line CCRF-CEM. AB - The tumor suppressor p53 has been implicated in apoptosis induction and is mutated in human T-ALL CCRF-CEM cells. To investigate possible consequences of wild-type p53 loss, we reconstituted CEM-C7H2, a subclone of CCRF-CEM, with a temperature-sensitive p53 allele (p53ts). Stably transfected lines expressed high levels of p53ts and shift to the permissive temperature (32 degrees C) caused rapid induction of p53-regulated genes, such as p21(CIP1/WAF1), mdm-2 and bax. This was followed by extensive apoptosis within 24 h to 36 h, supporting the notion that mutational p53 inactivation contributed to the malignant phenotype. p53-dependent apoptosis was preceded by digestion of poly(ADP-ribose) polymerase, a typical target of interleukin-1beta-converting enzyme (ICE)-like proteases/caspases, and was markedly resistant to the ICE/caspase-1 and FLICE/caspase-8 inhibitor acetyl-Tyr-Val-Ala-Asp.chloromethylketone (YVAD), but sensitive to the CPP32/caspase-3 inhibitor benzyloxycarbonyl-Asp-Glu-Val Asp.fluoromethylketone (DEVD) and benzyloxycarbonyl-Val-Ala Asp.fluoromethylketone (zVAD), a caspase inhibitor with broader specificity. This indicated an essential involvement of caspases, but argued against a significant role of ICE/caspase-1 or FLICE/caspase-8. Actinomycin D or cycloheximide prevented cell death, suggesting that, in this system, p53-induced apoptosis depends upon macromolecule biosynthesis. Introduction of functional p53 into CEM cells enhanced their sensitivity to the DNA-damaging agent doxorubicin, but not to the tubulin-active compound vincristine. Thus, mutational p53 inactivation in ALL might entail relative resistance to DNA-damaging, but not to tubulin destabilizing, chemotherapy. PMID- 9395241 TI - A novel epithelial-expressed ETS gene, ELF3: human and murine cDNA sequences, murine genomic organization, human mapping to 1q32.2 and expression in tissues and cancer. AB - The ETS family of genes are implicated in cancers such as Ewings sarcoma, acute myeloid leukemia and chronic myelomonocytic leukemia. Further, they have important functions in embryonic development. Hence, identification and characterization of members of this family are important. We identify a novel ETS family member, ELF3, and report its human and murine cDNA sequences. The mouse cDNA has an alternatively spliced transcript with an extra 60 bp inserted. Hence we present the organization of the murine Elf3 gene together with its exon/intron structure. This gene consists of 9 exons and 8 introns spanning 4.8 kb. ELF3 binds and transactivates ETS sequences and interestingly also shows the ability to bind a GGAT-like purine core, a preferential ETS1/ETS2 type binding site. The expression of ELF3, unlike most other ETS family members, is absent in hematopoietic cells and hematopoietic organs in humans and mice. Intriguingly, the gene is specifically expressed in cell lines of epithelial origin and in organs such as lung, stomach, intestine, kidney that have specialized epithelial cells. We localize the human gene to 1q32.2, a region that is amplified in epithelial tumors of the breast, lung and prostate. Finally, we show that ELF3 expression is increased in a lung carcinoma and adenocarcinoma, as compared to normal tissue. ELF3 is also expressed in cell lines derived from lung cancers. These results suggest that this novel ETS gene may be involved in lung tumorigenesis. PMID- 9395243 TI - The human protein p19ARF is not detected in hemopoietic human cell lines that abundantly express the alternative beta transcript of the p16INK4a/MTS1 gene. AB - The p16/MTS1/CDKN2 gene on human chromosome band 9p21 encodes two unrelated proteins: p16INK4a, a specific inhibitor of the cyclin D-dependent kinases CKD4 and CDK6, and the structurally unrelated p19ARF protein that arrests cell growth in G1/S and also in G2/M. By use of polyclonal antibodies, the human p19ARF (hp19ARF) protein has been identified in the nucleus of various cells including normal cultured fibroblasts. The level of this protein did not fluctuate throughout the cell cycle and was more elevated in fibroblasts with limited or arrested growth, suggesting that p19ARF accumulated in presenescent or senescent cells. Interestingly, hp19ARF was not detected in several hemopoietic tumor cell lines (mainly of B-type lymphoid origin) that expressed abundant amounts of the p16beta transcript. This finding indicates that in certain tumors, the expression of hp19ARF RNA and protein may be uncoupled. Furthermore, it suggests that disruption of a translational mechanism may be involved in the inactivation of hp19ARF. PMID- 9395242 TI - Cloning of two candidate tumor suppressor genes within a 10 kb region on chromosome 13q14, frequently deleted in chronic lymphocytic leukemia. AB - Previous studies have indicated the presence of a putative tumor suppressor gene on chromosome 13q14, commonly deleted in patients with B-cell chronic lymphocytic leukemia (B-CLL). We have previously defined a minimally deleted region of 130 kb centromeric to the marker D13S272, and constructed a PAC and cosmid contig encompassing this area. In the present study we have made a detailed restriction and transcriptional map of the region of interest. Using these tools we have screened a panel of 206 primary CLL clones and three cell lines. In five CLL cases we found limited deletions defining the region of interest to an area of no more than 10 kb. Two adjacent genes, termed Leu1 and Leu2 (leukemia-associated gene 1 and 2), were mapped to the minimally deleted region, with several patients showing deletion borders within these genes. The Leu1 and Leu2 genes show little homology to previously published genes at the nucleotide and expected translated amino acid sequence level. Mutational analysis of the Leu1 and 2 genes in 170 CLL samples revealed no small intragenic mutations or point mutations. However, in all cases of 13q14 loss examined, the first exon of both genes, which are only 300 bp apart, were deleted. We conclude that the Leu1 and Leu2 genes are strong candidates as tumor suppressor gene(s) involved in B-CLL leukemogenesis. PMID- 9395244 TI - Phosphorylated retinoblastoma protein stimulates DNA polymerase alpha. AB - Human retinoblastoma (Rb) protein, immunopurified from an extract of recombinant baculovirus infected cells, stimulated 10-100-fold the activity of DNA polymerase alpha from calf thymus or human HeLa cells. Purified Rb protein is composed of two electrophoretically distinguishable forms, i.e., partially phosphorylated and under-phosphorylated forms. Dephosphorylation of Rb protein by protein phosphatase 2A largely diminished its stimulatory effect. On the other hand, a hyperphosphorylated Rb protein, obtained from insect cells overexpressing Rb protein, cyclin E and cyclin-dependent kinase 2 simultaneously, stimulated DNA polymerase alpha more strongly than the singly-expressed Rb protein. These results indicate that the phosphorylation is crucial for the stimulation. Rb protein isolated from human Burkitt lymphoma Raji cells also stimulated DNA polymerase alpha. In contrast, Rb protein did not affect eukaryotic DNA primase or Klenow fragment of Escherichia coli DNA polymerase I. By immunoprecipitation using anti-DNA polymerase alpha antibody, Rb protein in nuclear extract of Raji cells was co-precipitated with DNA polymerase alpha. This result indicates that DNA polymerase alpha exists as a complex containing phosphorylated Rb protein in cells. DNA polymerase alpha specifically bound to a purified Rb protein immobilized Sepharose column. Rb protein also bound to DNA polymerase alpha trapped to anti-DNA polymerase alpha antibody-Sepharose column, suggesting the direct association of these two proteins. These observations suggest a new function of phosphorylated Rb protein in the regulation of DNA replication. PMID- 9395245 TI - Cross-talk in signal transduction: Ras-dependent induction of cAMP-responsive transcriptional repressor ICER by nerve growth factor. AB - The CREM gene encodes both activators and repressors of cAMP-induced transcription. By virtue of an alternative, intronic promoter within the gene, the ICER (Inducible cAMP Early Repressor) isoform is generated. ICER acts as a dominant negative regulator and is cAMP-inducible in various neuroendocrine cells and tissues. ICER negatively autoregulates its own expression, and appears to participate in the molecular events governing oscillatory hormonal regulations. Here we report that ICER is inducible with nerve growth factor (NGF). This is the first example of cAMP-independent induction of ICER expression. Importantly, induction by NGF occurs via a subset of the CREs present in the ICER promoter which were previously shown to direct cAMP-inducibility. ICER induction correlates with a NGF-mediated phosphorylation of CREB. Both CREB phosphorylation and ICER inducibility require an intact Ras-dependent signalling pathway. We show that increased ICER levels result in the attenuation of c-fos expression. The activation of a powerful repressor of cAMP-responsive transcription by NGF, whose transduction signalling is cAMP-independent, constitutes a notable example of nuclear cross-talk and thus is likely to have relevant physiological implications. PMID- 9395246 TI - DNA-damage-inducible p53 activity in SV40-transformed cells. AB - The biological state of the tumour suppressor proteins Rb and p53 is altered in papillomavirus- and SV40-transformed cells, due to interaction with the DNA tumour virus oncogene proteins E6/E7 and the tumour (T) antigen. Thus, the DNA damage response function of p53, a crucial feature of the tumour suppressor p53, might be considered as inactive. To investigate this subject, C57SV and VLM, two SV40-transformed murine cell lines enharboring constitutively high nuclear p53 and SV40 large T antigen levels, were treated with mitomycin C. Mitomycin C is known for its activity to elicit DNA damage, followed by nuclear accumulation of biologically active p53. Surprisingly, the nuclear p53 level significantly increased in mitomycin-C-treated C57SV cells and to a lesser degree in VLM cells. In addition, expression of p21WAF1 protein was induced in C57SV and VLM cells. This indicates a possible DNA-damage-elicited p53 activation. Finally, nuclear extracts of mitomycin-C-treated C57SV and VLM cells, but not of untreated cells, exhibited PAb421-enhanced specific DNA-binding activity of p53, as proven by gel shift analysis. Thus, DNA damage induced essential biological functions typical for wild-type p53 in the SV40-transformed cell lines examined so far. PMID- 9395247 TI - Interdomain binding mediates tumor growth suppression by the NF2 gene product. AB - The neurofibromatosis 2 (NF2) tumor suppressor gene encodes an intracellular membrane-associated protein, called merlin (or schwannomin), that belongs to the band 4.1 family of cytoskeleton-associated proteins. Inactivating NF2 mutations occur in several sporadic tumor types and have been linked to the NF2 disease, whose hallmark is the development of bilateral Schwann cell tumors (schwannomas) of the eighth cranial nerve. Two major alternatively spliced NF2 variants are expressed in normal tissues: 'NF2-17' lacking exon 16 and 'NF2-16' that contains exon 16 and encodes a merlin protein truncated at the C-terminus. We report that overexpression of NF2-17 in rat schwannoma cells inhibits their growth in vitro and in vivo, while NF2-16 fails to influence schwannoma growth. Tumor growth inhibition by merlin depends on an interdomain association occurring either in cis or in trans between the N- and C-termini. This association does not occur in the truncated NF2-16 protein nor in a mutant NF2-17 protein lacking C-terminal sequences. These data indicate that merlin has a unique mechanism of tumor suppression, inhibiting cell proliferation via self-association. PMID- 9395248 TI - Nitric oxide and inflammatory arthritides. AB - Nitric oxide (NO), first identified as endothelium-derived relaxing factor (EDRF), is a free radical synthesized from L-arginine by NO synthases (NOS). NO plays vital roles in biological responses, including regulation of vascular tone, neurotransmission, anti-viral defense and immune response. There are two isoforms in NOS; constitutive NOS (cNOS) and inducible NOS (iNOS). Inflammatory cytokines such as interleukin-1(IL-1), interferon-gamma (IFN-gamma), tumor necrosis factor alpha (TNF-alpha) induce iNOS expression in various cells including macrophages. NO production is increased in inflammatory arthritides both in rodent models and human. Enhanced NO production is observed in various compartment in vivo but inflammatory synovium and cartilage are the major source of NO. The onset of arthritis in rodent models is successfully blocked by the NOS inhibitor, NG monomethyl-L-arginine (L-NMMA). These data suggest a possible involvement of NO in the pathogenesis and tissue destruction in arthritis, and the significance of up-regulated NO production is discussed. PMID- 9395249 TI - Brain pharmacokinetics and in vivo receptor binding of 1,4-dihydropyridine calcium channel antagonists. AB - Brain pharmacokinetics of 1,4-dihydropyridine (DHP) calcium channel antagonists and their in vivo receptor binding in mice were characterized. The area under the concentration vs time curve (AUCbrain) for [3H]nifedipine, [3H]nimodipine and [3H]PN 200-110 in mouse brain after intravenous injection was higher than that for [3H]amlodipine. Brain/plasma concentration ratios (AUCbrain/AUCplasm) for [3H]nimodipine and [3H]PN 200-110 were 3 to 5 times higher than those for [ H]nifedipine and [3H]amlodipine. Further, brain/heart concentration ratios (AUCbrain/AUCheart) for [3H]nifedipine, [3H]nimodipine and [3H]PN 200-110 were about 20 times higher than the ratio for [3H]amlodipine. A significant amount of specific binding in particulate fractions of mouse brain was detected in vivo by intravenous injection of [3H]nifedipine, [3H]nimodipine and [3H]PN 200-110 but not [3H]amlodipine. These data suggest that [3H]nifedipine, [3H]nimodipine and [3H]PN 200-110 are more extensively taken up into brain from plasma than [3H]amlodipine and bind to the receptor sites in brain parenchymal cells in a significant amount in vivo. In conclusion, the present simultaneous measurement of pharmacokinetics and in vivo receptor binding in mouse brain suggests an usefulness of calcium channel antagonists such as nimodipine in the pharmacotherapy of brain diseases. PMID- 9395250 TI - Prothymosin alpha promotes cell proliferation in NIH3T3 cells. AB - Thymic hormones have immunomodulatory effects on T cells and hence have been used clinically to restore the immunity of immunodeficient patients as well as to enhance the cellular immunity of cancer patients. Prothymosin alpha, which is a member of the thymic hormone family, has recently been suggested to act as a nuclear protein participating in the stimulation of cell proliferation. To characterize the biological activities ofprothymosin alpha in vitro, we established NIH3T3 cell transformants that constitutively express higher prothymosin alpha protein and its mRNA compared with the wild-type counterpart. Cells that overexpressed prothymosin alpha increased the proliferative activity assayed by the [3H]-thymidine incorporation or by the cell cycle analysis with the fluorescent-activated cell sorter. The results provide direct evidence that prothymosin alpha plays a role in cell proliferation by shortening the duration of the G1 phase. PMID- 9395251 TI - Comparative study of radical scavenger and antioxidant properties of phenolic compounds from Vitis vinifera cell cultures using in vitro tests. AB - Vitis vinifera cell suspensions were used to isolate and characterize the flavonoids (anthocyanins, catechins) and non-flavonoids (stilbenes) found in red wine. Furthermore, we showed that astringin is produced although this stilbene has not previously been reported to be a constituent of V. vinifera or wine. The ability of these compounds to act as radical scavengers was investigated using 1,1-diphenyl-2-picryl-hydrazyl (DPPH), a stable free radical. Antioxidant activities were assessed by their capacity to prevent Fe2+-induced lipid peroxidation in microsomes and their action on Cu2+-induced lipid peroxidation in low-density lipoproteins. The results showed that astringin has an important antioxidant effect similar to that of trans-resveratrol, and a higher radical scavenger activity than the latter. Astringinin appeared to be more active. These data indicate that phenolic compounds (stilbenes, catechins, anthocyanins) exhibit interesting properties which may account in part for the so-called "French paradox," i.e. that moderate drinking of red wine over a long period of time can protect against coronary heart disease. PMID- 9395252 TI - Expression of prostaglandin EP2 receptor mRNA in the rat spinal cord. AB - RT-PCR and an in situ hybridization analyses demonstrated expression of prostaglandin EP2 receptor mRNA in the subfield of the dorsal horn of the rat spinal cord. The intensive signals from the cells labeled with digoxigenin-11 dUTP were scattered through this region and they corresponded to neurons. EP2 receptor-mediated signal transduction is expected to play roles in modulating some neuronal functions such as transmission of nociceptive reaction or motor reflex. PMID- 9395253 TI - Characterization of LY344864 as a pharmacological tool to study 5-HT1F receptors: binding affinities, brain penetration and activity in the neurogenic dural inflammation model of migraine. AB - LY344864 is a selective receptor agonist with an affinity of 6 nM (Ki) at the recently cloned 5-HT1F receptor. It possesses little affinity for the 56 other serotonergic and non-serotonergic neuronal binding sites examined. When examined for its ability to inhibit forskolin-induced cyclic AMP accumulation in cells stably transfected with human 5-HT1F receptors, LY344864 was shown to be a full agonist producing an effect similar in magnitude to serotonin itself. After an intravenous dose of 1 mg/kg, rat plasma LY344864 levels declined with time whereas brain cortex levels remained relatively constant for the first 6 hours after injection. Oral and intravenous LY344864 administration potently inhibited dural protein extravasation caused by electrical stimulation of the trigeminal ganglion in rats. Taken together, these data demonstrate that LY344864 is a selective 5-HT1F receptor agonist that can be used to explore both the in vitro and in vivo functions of this receptor. PMID- 9395254 TI - Comparison of the Ca2+ movement by activation of alpha1-adrenoceptor subtypes in HEK-293 cells. AB - We studied the Ca2+ movement induced by activation of alpha1A-, alpha1B- and alpha1D-adrenoceptor subtypes in transfected HEK-293 cells with the fura-2 probe. All these alpha1-AR subtypes induced both Ca2+ release and Ca2+ entry. The effect on Ca2+ release in alpha1b transfected HEK-293 cells was bigger than that in alpha1a and alpha1d transfected HEK-293 cells, and the effects on Ca2+ entry were the same in alpha1a, alpha1b and alpha1d transfected HEK-293 cells. The Ca2+ entry was inhibited by 1 mM NiSO4, but not by nifedipine. Cyclopiazonic acid (CPA) produced a biphasic Ca2+ signal response in Ca2+ medium, and only induced a transient response in Ca2+-free medium. After depletion of CPA-sensitive Ca2+ pool by 10 microM CPA in Ca2+-free medium, 10 microM adrenaline (Adr) still transiently increased [Ca2+]i in three different alpha1-adrenoceptor subtype transfected HEK-293 cells. However, after depletion of adrenaline-sensitive Ca2+ pool by 10 microM Adr, CPA transiently elevated [Ca2+]i only in alpha1a and alpha1d transfected HEK-293 cells, not in alpha1b transfected HEK-293 cells. U73122, a phospholipase C (PLC) inhibitor, inhibited both Ca2+ release and Ca2+ entry induced by activation of alpha1A alpha1B and alpha1D subtypes in transfected HEK-293 cells. These results suggest that HEK-293 cell line contains two functionally separate intracellular Ca2+ pools, CPA-sensitive and Adr sensitive pools. Activation of alpha1B-AR stimulates Ca2+ release from both CPA sensitive and Adr-sensitive Ca2+ pools. Alpha1A and alpha1D subtypes induce Ca2+ release only from Adr-sensitive Ca2+ pool. PMID- 9395255 TI - Hemolytic effects of dehydroepiandrosterone in vitro. AB - Dehydroepiandrosterone (DHEA), a major steroid secreted by the adrenal gland which decreases with age after adolescence, is available as a over-the-counter product. This study demonstrates that DHEA induced lysis of human red blood cells (RBCs) in a concentration-dependent manner, with ca. 70% hemolysis at 2 mM DHEA at 37 degrees C for 1 hr. Hemolysis induced by 2 mM DHEA was rapid and involved neither hemoglobin oxidation nor lipid peroxidation. The hemolysis was also not inhibited by addition of EDTA, catalase, superoxide dismutase, glucose or a radical scavenger including mannitol, dimethylsulfoxide and alpha-tocopherol, indicating a non-oxidative mechanism. RBCs stored overnight before incubation with DHEA were hemolyzed to a lesser extent than the freshly prepared RBCs. Light microscopy of the fresh RBCs following 1-h incubation with 2 mM DHEA revealed thickened and cup-shaped deformity of the membranes, suggesting a change in the membrane structure possibly due to the intercalation of the steroid into the membranes. PMID- 9395257 TI - Theoretical analysis of a morphine withdrawal phenotype in a cultured cell line. AB - We have previously described a delta-opioid receptor-expressing cultured cell line that proliferates in a defined medium and responds to chronic morphine treatment with an inhibition of its rate of proliferation. To help provide an explanation for this behavior, we have used computer simulation of cell cycle kinetics to analyze the observed rates of proliferation of these cells in the presence and absence of morphine, and after withdrawal of morphine treatment. We questioned whether the difference in cell kinetics observed for the cell populations under the different treatments could be due to changes in the length of the cell cycle, withdrawal of cells from the cycle into a quiescent state, or differences in cell renewal. This was investigated by comparing observed cell numbers as a function of time with the results of different computer simulations using different values for these parameters. We found that we can provide a satisfactory explanation of the experimental observations on the basis of changes in a small set of parameters: Untreated cells experience a slowdown of cell proliferation at about the culture density where multiple cell-cell contacts are made and, beginning then, a large fraction are shunted from G1 into a quiescent state. Chronic morphine treatment inhibits proliferation by slowing passage through G1, but the cells remain as sensitive to cell-cell contacts as the untreated cells. After drug withdrawal following a 6 day treatment with morphine, the cells exhibit a large temporary increase in their rate of proliferation compared with control or chronically treated cells but about 48 hours after withdrawal, when cell-cell contacts just begin to be made, the cells return to almost their pre-treatment total cell cycle time and, as before, a large fraction are shunted into a quiescent state. Taken in conjunction with previously published results, the present ones indicate a possible interaction between morphine-induced and insulin-induced nuclear signaling pathways to the nucleus. PMID- 9395256 TI - Serotonin transporter gene-linked polymorphic region: allele distributions in relationship to body weight and in anorexia nervosa. AB - Several lines of evidence implicate a role for the serotonergic system in body weight regulation and eating disorders. The magnitude and duration of postsynaptic responses to serotonin (5-HT) is directed by the transport into and release from the presynaptic neuron. Recently, a common polymorphism of a repetitive element in the region of the serotonin transporter (5-HTT) gene-linked polymorphic region (5-HTTLPR) was identified that results in a system of two common alleles. The activity of the 5-HTT, as measured in in vitro assays and in human lymphoblastoid cell lines, is dependent on the respective genotype. We thus hypothesized that this polymorphism is relevant for weight regulation in general and is possibly involved in the etiology of anorexia nervosa (AN). Allele frequencies and genotypes were determined in a total of 385 unrelated obese children, adolescents and adults, 112 underweight subjects and 96 patients with AN. Furthermore, both parents of 98 obese children and adolescents and of 55 patients with AN, respectively, were genotyped, thus allowing to test for both association and linkage. The comparison of allele frequencies between obese and underweight probands provided no evidence for a major role of the 5-HTTLPR in weight regulation. Patients with AN had allele frequencies not significantly different to those observed for obese and underweight individuals. PMID- 9395258 TI - Effect of Ca2+-channel blockers on isoprenaline-induced desensitization in rat trachea. AB - Isoprenaline-induced desensitization in vitro in rat trachea was studied in the presence of the Ca2+-channel blockers (Ca2+-CBs) verapamil and nitrendipine. The concentration-response curves for isoprenaline were determined in a noncumulative manner using carbachol as contracting agent, and then desensitization was achieved by 40-min incubation of the tracheal preparations with isoprenaline (1 microM). The effect of verapamil and nitrendipine was studied by the addition of each Ca2+-CB to the desensitizing solution. Both verapamil and nitrendipine reduced the isoprenaline-induced desensitization in the rat trachea. PMID- 9395259 TI - Interaction mechanisms of imipramine and desipramine with enkephalin-degrading aminopeptidases in vitro. AB - In the last few years, considerable evidence has appeared concerning the importance of the opioid systems in the action mechanism of some antidepressant drugs. This action mechanism could be mediated through the inhibition of the enzymes responsible for enkephalin degradation. In this sense, imipramine treatment in vivo increases the enkephalin levels, and this effect is enhanced by inhibitors of enkephalin-degrading enzymes. The present work shows the effects in vitro of imipramine and its active metabolite desipramine on the activities of two membrane-bound enkephalin-degrading aminopeptidases present in rat brain. Imipramine and desipramine in vitro do not affect the aminopeptidase M activity, but they reversibly inhibits the aminoeptidase MII. The enzyme kinetic analysis shows that this enzyme molecule has two different binding sites for each drug, which exert a mixed type enzyme inhibition. PMID- 9395260 TI - Induction of the human sperm acrosome reaction with mannose-containing neoglycoprotein ligands. AB - In the interest of classifying cases of male factor infertility, we have paid particular attention to the sugar ligand binding properties of the human sperm surface and the functional capacity of the acrosome for exocytosis--key parameters for assessing sperm fertilizing ability. Zona recognition and binding involve the interactions of sperm surface mannose receptors (lectins) with mannose ligands on the zona pellucida. Sperm surface mannose lectins can be visualized by their ability to bind a synthetic model zona ligand, fluorescein isothiocyanate (FITC)-conjugated mannosylated bovine serum albumin (BSA) (Man FITC-BSA). We now report that Man-FITC-BSA biologically also mimics the effects of solubilized authentic human zonae, in that binding of Man-FITC-BSA results in a time-dependent receptor aggregation and the induction of acrosome exocytosis in capacitated sperm populations from fertile donors. In our assay, the addition of mM amounts of mannose monosaccharide to Man-FITC-BSA increases the number of polyvalent mannose ligands bound by individual spermatozoa and increases the rate of the acrosome reactions induced by Man-FITC-BSA, thereby increasing specimen processing efficiency. We conclude that exposure of human spermatozoa to polyvalent mannose ligands + D-mannose monosaccharide offers a new, convenient and readily available system to study sperm capacity for induced acrosome loss. PMID- 9395261 TI - Use of mannose ligands in IVF screens to mimic zona pellucida-induced acrosome reactions and predict fertilization success. AB - A predictive test for determining whether motile populations of human spermatozoa will fertilize eggs in vitro has been an elusive goal of clinical research. We have developed an assay for the ability of motile human spermatozoa to bind fluorescein isothiocyanate-labelled mannosylated bovine serum albumin (Man-FITC BSA) as a test for the presence of sperm surface receptors (lectins) for mannose ligands. Mannosylated ligands are present on the human zona pellucida and are involved in the species-specific binding of human spermatozoa to the zona pellucida. We now demonstrate in prospective blinded analysis that the fractional increase in acrosome loss following a mannose ligand challenge is highly correlated with the rate of fertilization in vitro. Using an incremental increase of acrosome exocytosis of >0.1 as a threshold to predict which specimens will yield normal fertilization, the assay has a sensitivity of 97.8%, a specificity of 83.3%, a positive predictive value of 95.7% and a negative predictive value of 90.7%. These data indicate that testing for a mannose-induced acrosome reaction may be useful in assessment of sperm function prior to in-vitro fertilization in order to assign males to conventional insemination or intracytoplasmic sperm injection protocols. PMID- 9395262 TI - Assisted reproductive technology and complex chromosomal rearrangements: the limits of ICSI. AB - Complex chromosomal rearrangements are very rare events in the human population. According to our knowledge on the consequences of simple reciprocal translocations for male fertility, translocations involving three or more chromosomes are thought to lead to severe reproductive impairments in terms of meiotic disturbance or chromosomal imbalance of gametes. We report the case of a 48 year old man whose sperm count revealed either oligozoospermia (<10(3) spermatozoa/ml) or azoospermia. He was referred to the laboratory for in-vitro fertilization after intracytoplasmic sperm injection. Cytogenetic investigations showed a complex chromosomal rearrangement involving firstly a translocation between the short arm of chromosome 7 and the long arm of chromosome 13 and secondly a translocation between the short arm of the same chromosome 13 and the short arm of chromosome 9. Diagnosis was ascertained by fluorescence in-situ hybridization and staining of the nucleolar organizer regions. Theoretical study of the translocated chromosomes predicted a 'chain' configuration of the hexavalent at the pachytene stage of meiosis. In all, 32 modes of segregation were considered and only one resulted either in a normal or a balanced gamete karyotype. Genetic counselling and choice of appropriate artificial reproduction technique are discussed. PMID- 9395264 TI - Oocyte polarity and cell determination in early mammalian embryos. AB - Knowledge on determination and differentiation in the mammalian embryo has not kept pace with discoveries in other phyla. Current concepts overlook well established pathways leading to polarity in oocytes and embryos of other phyla, modern principles of totipotency in plants and animals, and axis formation in lower vertebrates. Various models derived from invertebrates and frogs could be relevant to the situation in eutherian mammals, and we explore the nature of strict genetic controls in these species and its implications for early mammalian differentiation. Concepts on totipotency and related phenomena in animal and human embryos are examined and the possibility raised that two cell lines are formed in early human embryos from the 2-4 cell stage. Clinical consequences are assessed, including causes of the high incidence of chromosomal mosaicism in human embryos. Our interpretations are obviously speculative, and must be clarified by experimentation. PMID- 9395263 TI - Analysis of P[gal4] insertion lines of Drosophila melanogaster as a route to identifying genes important in the follicle cells during oogenesis. AB - We report the analysis of a number of lines of Drosophila melanogaster containing insertions of the yeast gal4 gene. By crossing a UAS-lacZ fusion gene as a reporter into these lines, we analysed the expression patterns of beta galactosidase during oogenesis. Since there is no expression of GAL4 in the germ line in these experiments, this is an ideal system for the analysis of expression patterns in sub-sets of follicle cells. These lines provide ideal markers for sets of follicle cells, e.g. anterior or posterior polar cells for studying genetic interactions in oogenesis; however, they can also be used in the same way as conventional enhancer traps to clone nearby genes with similar expression patterns. The advantages of this dual gal4/UAS system over conventional enhancer trapping includes the possibility of GAL4-directed misexpression and antisense expression studies to establish the function of the genes we identified during follicle cell determination and differentiation. These studies could lead to the isolation of homologous genes crucial in mammalian oogenesis. Understanding how the somatic cells and germ cells interact to promote growth and maturation of the mammalian follicle and oocyte could well be crucial for improving the fertility of eggs used for in-vitro fertilization programmes, and could provide methods for assessing the quality of eggs. PMID- 9395265 TI - Cell surface carbohydrates and lectins in early development. AB - Current knowledge on the development regulation of cell surface carbohydrates and lectins in mammalian embryos is summarized. Much of this data comes from observations on mouse embryos but information on the human embryo is included where it is available. Over the last few years, numerous studies have indicated that carbohydrates play a critical role in the cell-cell interactions of the pre- and peri-implantation embryo. Functional tests suggest a role for terminal fucosylated Galbeta1-3/4GlcNAc structures in the early steps of implantation. We also now have clear evidence for the expression of lectins in the trophectoderm just prior to implantation. Mouse mutants have been generated which lack particular enzymes involved in glycosylation or particular lectins, but so far they have not been informative about the role of glycoconjugates or lectins in the very early embryo. PMID- 9395266 TI - Cell death in the mammalian blastocyst. AB - Cell death is a widespread feature in the blastocysts of many mammals. Isolated cells in both the inner cell mass and the trophectoderm undergo cell death. These dying cells appear morphologically to be undergoing apoptosis. In mouse blastocysts, a wave of cell death is seen in vivo, suggesting that it plays an important role in normal development. However, cell death is increased under suboptimal culture conditions. There is evidence that levels of cell death are regulated by 'survival' factors produced both by the embryo itself and by the maternal reproductive tract. The role of cell death in development is unknown, but could involve the elimination of abnormal cells, or a sublineage of cells with an inappropriate developmental potential. Work in other systems has demonstrated that cell death is regulated by the activity of apoptosis genes. Whether these genes are implicated in blastocyst cell death, and the reasons for apoptosis in the early embryo, remain to be determined. PMID- 9395267 TI - Nikolaj Nikolajewitsch Anitschkow (1885-1964) established the cholesterol-fed rabbit as a model for atherosclerosis research. AB - The cholesterol-fed rabbit is a widely used model for experimental atherosclerosis research. In regard to this, one name is periodically mentioned: Nikolaj Nikolajewitsch Anitschkow. Those infrequent reminders of an important name in modern medical history do not pay an adequate tribute to basic findings concerning the pathology and pathogenesis of atherosclerosis. In contrast to research groups at that time conducting experiments with protein enriched diets, Anitschkow demonstrated, in 1913 in St. Petersburg, that it was cholesterol only that caused these atherosclerotic changes in the rabbit arterial intima, which was very similar to human atherosclerosis. By analysing the plaque's development and histology, Anitschkow was able to identify the cell types, on which modern atherosclerosis research is now focussing with a new set of immunohistochemical methods: smooth muscle cells, macrophages and lymphocytes. He noted early (fatty streaks) and advanced (atheromatous plaques) lesions and, by standardizing cholesterol feeding, he discovered that the amount of cholesterol uptake was directly proportional to the degree of atherosclerosis formation. His explanation for this observation was what modern terminology calls 'response-to-injury'. With modern immunohistochemical and molecular-biological methods, the cholesterol-fed rabbit can be used to investigate the pathophysiological aspects which also contribute to human atherosclerosis, such as lipoproteins, diabetes, mitogens, growth-factors, adhesion molecules, endothelial-function, receptor-pathways or platelets. This model can be combined with a number of other methods causing endothelial dysfunction and injury, such as balloon denudation, electric stimulation, cuff implantation, artificial hypertension, diabetes or infection. Bred strains of hereditary hypercholesterolemic rabbits or those resistant to a cholesterol-diet provide further possibilities to expand experimental designs. PMID- 9395268 TI - Endothelial-derived nitric oxide preserves anticoagulant heparan sulfate expression in cultured porcine aortic endothelial cells. AB - Nitric oxide (NO) has been shown to inhibit platelet adhesion and aggregation, but there are no reports on its interaction with the coagulation system. We investigated the effect of the L-arginine analogues, N-nitro-L-arginine (LNA), N(G)-nitro-L-arginine methyl ester (L-NAME), and N(G)-monomethyl-L arginine (L NMMA), competitive inhibitors of NO production, on endothelial-surface heparan sulfate. Addition of LNA to porcine aortic endothelial cells reduced 125I-labeled antithrombin III binding to the cell surface heparan sulfate in a dose- and time dependent fashion. Significant inhibition was observed with 1 mM LNA, and the maximal suppression (-50% of control) occurred at 10 mM LNA after 12 h. L-NAME (1 mM) and L-NMMA (1 mM) also significantly inhibited the antithrombin III binding. The iron chelator desferrioxamine significantly prevented the reduction of antithrombin III binding to LNA-treated cells. We further investigated the effect of L-NAME on intracellular oxidative stress of endothelial cells using a hydroperoxide-sensitive fluorochrome, carboxy-dichloro-dihydrofluorescein diacetate bisacetoxymethyl ester probe, and revealed that inhibition of NO synthesis by L-NAME led to a marked increase in intracellular oxidative stress. These results demonstrated that the prolonged inhibition of NO synthesis in porcine aortic endothelial cells decreases the expression of anticoagulant heparan sulfate on endothelial cells through the increase in intracellular oxidative stress, perhaps comprising another mechanism by which NO affects the coagulation system in the vasculature. PMID- 9395270 TI - Analysis of the T cell receptor V beta repertoire in human aortic aneurysms. AB - Human aortic aneurysm is commonly characterized by the presence of advanced atherosclerosis associated with variable chronic adventitial inflammation. Histological examination of human aortic aneurysmal specimens revealed the presence of plasma cells and lymphoid aggregates in media and adventitia of the vessels. Immunostaining further demonstrated that CD3-positive T lymphocytes are present in follicles. Using a highly sensitive reverse transcription-polymerase chain reaction amplification method, the T cell receptor (TCR) V beta gene expression in aortic aneurysms was shown to be polyclonal. Furthermore. there was no preferential expression of any TCR V beta gene in the aortic tissue as compared with that in peripheral blood in aneurysmal patients. These results indicate that the TCR repertoire in aortic aneurysm is not restricted. PMID- 9395269 TI - Subendothelial smooth muscle cells of human aorta express macrophage antigen in situ and in vitro. AB - Cells bearing a smooth muscle cell marker--alpha-actin and a macrophage marker- CD68 antigen were immunocytochemically identified on 'en face' preparations of human aortic intima. Cells, expressing smooth muscle alpha-actin, macrophage CD68 antigen and both markers, i.e. smooth muscle cells possessing the macrophage antigen, were identified both in grossly normal aortic areas and in atherosclerotic lesions (fatty streaks and atherosclerotic plaques). CD68 positive smooth muscle cells were most common in the lipid-rich areas: fatty streaks and atherosclerotic plaque shoulders. Cells expressing smooth muscle alpha-actin and CD68 were also revealed in primary cultures prepared from grossly normal and atherosclerotic intima. Cells expressing both antigens were found in all examined cultures. The proportion of these cells in cultures from grossly normal areas and atherosclerotic plaques was similar: 14.5 +/- 4.1 and 14.6 +/- 4.8%, respectively. Cultures from fatty streaks had a higher content of cells expressing both antigens: 25.1 +/- 7.0%. Modified low density lipoprotein-induced intracellular lipid accumulation in cells cultured from grossly normal intima led to a three-fold increase in the number of cells sharing alpha-actin and CD68 antigen. Accumulation of latex beads by phagocytosis had a similar effect. It was suggested that in atherosclerotic lesions intracellular lipid accumulation and other stimulators of phagocytosis may provoke the expression of macrophage associated antigen CD68 in settled cells of the subendothelial intima of human aorta. PMID- 9395271 TI - Effects of green tea, black tea and dietary lipophilic antioxidants on LDL oxidizability and atherosclerosis in hypercholesterolaemic rabbits. AB - The hypothesis that tea or dietary lipid-soluble antioxidants reduce atherogenesis by lowering the oxidizability of low-density lipoprotein (LDL) was investigated. Five groups of 20 female New Zealand white rabbits were fed a restricted amount of a high-fat (30 en%) semipurified diet supplemented with cholesterol (0.15%, w/w) for 21 weeks. The vitamin E content of the control diet was 40 mg/kg diet. The animals received either green tea or black tea in their drinking water or vitamin E (200 mg/kg diet) or beta-carotene (20 mg/kg). The serum cholesterol concentrations (in the order of 18-23 mmol/l) were not significantly different between the groups. Vitamin E was substantially increased as compared to controls in vitamin E supplemented animals (3-fold within 8 weeks in plasma and LDL; P < 0.01) and weakly (1.2-fold) by green and black tea (P < 0.05). Green tea consumption tended to reduce aortic lesion formation by 31% (24 +/- 3.2% versus 35 +/- 5.7% for control animals P = 0.11), while black tea, vitamin E and beta-carotene had no effect. This was in contrast to the resistance of isolated LDL to oxidation induced at high copper concentration. Green and black tea induced a 13% and 15% (P < 0.05) prolongation of the lag phase, respectively, with a correspondingly lower oxidation rate, while vitamin E increased the lag phase by 63% (P < 0.01) with a concomitant diminution of the oxidation rate and beta-carotene had no effect. Regression analysis showed that there was no relationship between the extent of atherosclerosis and LDL oxidizability or plasma malondialdehyde as marker of in vivo lipid peroxidation. The results of the present study raise the question whether LDL oxidizability (at least when tested at high induction rate ex vivo) is a primary causal mechanism in atherosclerosis in the cholesterol-fed rabbit. The suitability of the cholesterol-fed rabbit with extreme hypercholesterolaemia as a model to study antiatherosclerotic properties of dietary antioxidants, such as the tested polyphenols, is discussed. PMID- 9395272 TI - N(G)-nitro-L-arginine- and indomethacin-resistant endothelium-dependent relaxation in the rabbit renal artery: effect of hypercholesterolemia. AB - Studies were designed to compare the N(G)-nitro-L-arginine- and indomethacin resistant, endothelium-dependent relaxation to acetylcholine in isolated renal artery rings from normal and cholesterol-fed rabbits. It was assumed that the resistant part in response to acetylcholine is mediated by the endothelial derived hyperpolarizing factor (EDHF). Rabbits were fed normal (n = 15) or cholesterol enriched chow (n = 13, 1% cholesterol for 4 weeks, 0.5% for 12 weeks). In organ chamber experiments, renal artery rings were precontracted with 0.1-1 microM phenylephrine or 35 mM KCl, and relaxed with acetylcholine (0.001-10 microM) in the presence of 10 microM indomethacin. Studies were performed in the presence or absence of: 100 microM N(G)-nitro-L-arginine (L-NOARG) to inhibit the nitric oxide pathway, 100 nM charybdotoxin (CTX) or 1 mM tetrabutylammonium (TBA) to inhibit Ca2+-activated K+ channels, and 100 microM SKF 525a to inhibit cytochrome P450 monoxygenase pathway. In normal arteries, L-NOARG partially inhibited acetylcholine-induced relaxation. The resistant part was almost abolished when the arteries were depolarized with KCl, or when L-NOARG was combined with either CTX, TBA or SKF 525a. In arteries from hypercholesterolemic animals, the relaxation to acetylcholine was only slightly impaired as compared to normal animals. However, in comparison to arteries from normal animals, the L NOARG-resistant part of acetylcholine-induced endothelium-dependent relaxation was enhanced. It is speculated that differences in the balance between nitric oxide (NO)- and EDHF-mediated control of vascular tone may maintain acetylcholine induced vasodilatation of the renal artery in hypercholesterolemia. PMID- 9395273 TI - Beta-very low density lipoprotein induces triglyceride accumulation through receptor mediated endocytotic pathway in 3T3-L1 adipocytes. AB - To elucidate the mechanism of triglyceride (TG) accumulation in adipocytes induced by TG-rich lipoproteins, we examined the effect of beta-very low density lipoprotein (beta-VLDL) on TG accumulation in 3T3-L1 adipocytes. Beta-VLDL did not induce TG accumulation in 3T3-L1 preadipocytes but in 3T3-L1 adipocytes. TG accumulation was significantly inhibited by cytochalasin B, an inhibitor of receptor mediated endocytosis. In contrast, cytochalasin B did not inhibit free fatty acid induced TG accumulation in adipocytes. The binding of [125I]beta-VLDL to preadipocytes was inhibited completely by both beta-VLDL and LDL. In sharp contrast, the binding of [125I]beta-VLDL to adipocytes was inhibited completely by beta-VLDL, but partially by LDL. The VLDL receptor mRNA was only expressed in adipocytes. These results suggest that beta-VLDL induced TG accumulation in adipocytes may be mediated through the VLDL receptor pathway. PMID- 9395274 TI - Unrestricted usage of immunoglobulin heavy chain genes in B cells infiltrating the wall of atherosclerotic abdominal aortic aneurysms. AB - The aim of this study was to provide evidence for the hypothesis that the B cell rich infiltrate concentrated in the adventitia of atherosclerotic abdominal aortic aneurysms is an autoimmune response to specific tissue antigens. Detailed histological examination of biopsies from 26 atherosclerotic abdominal aortic aneurysms showed in the adventitia, the presence of lymphoid follicles in 7/26 (27%) and of plasma cells in all cases. DNA prepared from the outer aneurysm wall (n = 25) was amplified using the polymerase chain reaction to investigate the repertoire of the immunoglobulin heavy chain (VH) genes used. Amplification of the VDJ region of VH, using both framework 2 and 3 primers, revealed unrestricted usage of the VH gene in 24/25 cases. The only case where restricted usage of the VH genes was observed, might have been attributable to severe virally-induced tissue inflammation. These results indicate that, in the vast majority of atherosclerotic abdominal aortic aneurysms, the B cell rich adventitial infiltrates are not an autoimmune response to a limited repertoire of tissue antigens. PMID- 9395275 TI - The effect of hormone replacement therapy on the oxidation of low density lipoprotein in postmenopausal women. AB - The oxidative modification of low density lipoprotein (LDL) is important in the pathogenesis of atherosclerosis, and oestrogen has been shown to inhibit copper and cell mediated oxidation of LDL in vitro. We have investigated the effect of oral and transdermal oestrogens (oestradiol valerate 1 mg, conjugated equine oestrogens 0.625 mg and patches releasing 50 microg oestradiol daily), oestrogen implants (oestradiol 50 mg) and oral combined oestrogen and progestogen (oestradiol valerate 2 mg with medroxyprogesterone acetate 5 mg and oestradiol valerate 2 mg with norethisterone acetate 1 mg), on the susceptibility of LDL to oxidation in postmenopausal women (total n = 56). Oxidation of LDL was initiated by the addition of copper ions, and monitored by measurement of conjugated dienes. Changes in fasting serum levels of total cholesterol, LDL, HDL and triglycerides were also evaluated, as were changes in LDL composition. Total cholesterol decreased by 5.5% (P < 0.05) with CEE, 6.8% with oestradiol implants (P < 0.05), 9.3% with oestradiol + MPA (P < 0.01) and 10% with oestradiol + norethisterone (P < 0.05). There were reductions in LDL with oral oestradiol valerate (7.8%) (P < 0.05), CEE (13.8%) (P < 0.01) and oestradiol combined with MPA (12.7%) (P < 0.05). HDL increased by 7.1% (P < 0.01) and 6.3% (P < 0.05), with oestradiol valerate and CEE respectively, and decreased by 9% (P < 0.05) with implants and by 14.7% with oestradiol combined with norethisterone (P < 0.01). Triglycerides were significantly increased with CEE (14.9%) and reduced with oestradiol implants (15.2%) (both P < 0.05). While there was no change in the ratio of 'cholesterol ester' to 'free cholesterol' within LDL with any of the HRT preparations, a reduction in total cholesterol and cholesterol ester content of LDL occurred with transdermal oestradiol and a reduction in free cholesterol occurred with oestradiol plus MPA. Although we found a small but significant decrease in plasma hydroperoxide concentration four weeks after insertion of the oestradiol implant from 1.17 +/- 0.06 to 1.03 +/- 0.04 micromol/l (P < 0.05), we found no significant change in the lag time to oxidation, or in the maximum rate of propagation of the reaction, after treatment with any of the above forms of hormone replacement therapy. This study does not therefore support the role of oestrogens as antioxidants in vivo. PMID- 9395276 TI - Direct association between the hepatic secretion of very-low-density lipoprotein apolipoprotein B-100 and plasma mevalonic acid and lathosterol concentrations in man. AB - Apolipoprotein B-100 (apo B) is the principal structural and functional protein of the pro-atherogenic lipoproteins, but its homeostasis in man has not been clearly established. The hepatic availability of cholesterol substrate may be a determining factor. We examined whether there was a direct correlation between plasma concentrations of mevalonic acid (MVA) and lathosterol (indices of in vivo cholesterol synthesis) and hepatic secretion of very-low-density lipoprotein (VLDL) apo B in 13 normolipidaemic, healthy male subjects. The secretion of VLDL apo B was measured using a primed constant intravenous infusion of 1-[13C] leucine (1 mg/kg per h) over 8 h. Gas-chromatography mass spectrometry (GCMS) was used to derive isotopic enrichment of apo B and fractional turnover rate was calculated using a monoexponential function. There was a highly significant positive correlation between the absolute secretion rate (ASR) of VLDL apo B and the plasma concentrations of mevalonic acid (r = 0.72, P = 0.005) and lathosterol (r = 0.81, P = 0.001) and the lathosterol:cholesterol ratio (r = 0.79, P = 0.001). In multiple regression analysis, these correlations remained significant after adjusting for waist circumference, age, apolipoprotein E genotype and dietary fat intake. The data further support the notion that the availability of cholesterol substrate regulates the hepatic secretion rate of apo B. PMID- 9395277 TI - Red wine and fractionated phenolic compounds prepared from red wine inhibit low density lipoprotein oxidation in vitro. AB - The oxidative modification of low density lipoproteins (LDL) has been implicated in the development of atherosclerosis. This study examined the effect of red wine, ethanol and red wine stripped of phenols on copper-mediated and azo initiated LDL oxidation. Red wine containing phenolic compounds (0.025-20 mg/l gallic acid equivalents) increased the lag time of conjugated diene formation, inhibited the generation of thiobarbituric acid reactive substances (TBARS) and decreased the relative electrophoretic mobility of LDL in a concentration dependent manner. These changes were not apparent in LDL incubated with ethanol or red wine stripped of phenols. In other experiments, red wine (75 mg/l gallic acid equivalents) was incubated with plasma at 37 degrees C for 3 h. The LDL isolated from this plasma displayed a 60% increase in lag time following copper mediated oxidation. Uptake of this LDL by cultured J774 macrophages was three fold lower than control LDL. Red wine was fractionated into phenolic acids (fraction 1), catechins and monomeric anthocyanidins (fraction 2), flavonols (fraction 3) and polymeric anthocyanidins (fraction 4). All red wine fractions prolonged the time before LDL oxidation. Fraction 2 displayed a significantly greater antioxidant activity than fractions 3 and 4 (but not fraction 1) in at least one pro-oxidant model. In conclusion we have shown that antioxidant compounds in red wine can associate with LDL particles following an incubation in whole plasma, can exert an antioxidant effect and, in so doing, can inhibit the uptake of the lipoprotein by macrophages. This antioxidant effect of red wine was apparent in most of the phenolic fractions separated from wine, particularly catechins, monomeric anthocyanidins and phenolic acids. PMID- 9395278 TI - Lymphocytes in human atherosclerotic plaque exhibit the elastin-laminin receptor: potential role in atherogenesis. AB - The purpose of this immuno-histochemical study was to investigate if lymphocytes, present in the human atherosclerotic plaque, exhibit the elastin-laminin receptor. We showed recently that human activated lymphocytes in vitro express this receptor. Briefly, we demonstrated by immuno-localization experiments and by flow cytometry that this receptor is available on the cell surface of human activated lymphocytes, free to react with ligands and show capping. The activation of this receptor by elastin peptides triggers several cellular reactions of biological interest as shown previously such as chemotactic movement to an elastin peptide gradient, modulation of the biosynthesis of connective tissue macromolecules, increase of protease synthesis and release of free radicals (O2-., NO.) from mononuclear and endothelial cells, modifications of ion fluxes and also increase of cell proliferation. All these processes may contribute to the development of the atherosclerotic lesion. Two of the previously demonstrated cell reactions mediated by the receptor could be demonstrated also on PHA-stimulated human lymphocytes namely stimulation of cell proliferation and increase of elastase activity. We demonstrated in the present immuno-histological study that about 50-60% of lymphocytes of the human atherosclerotic plaque obtained by endarterectomy express the 67 kDa subunit of the elastin-laminin receptor confirming that the above described phenomena could contribute to the chronicity of the lesion. PMID- 9395280 TI - Cerivastatin: pharmacology of a novel synthetic and highly active HMG-CoA reductase inhibitor. AB - The pyridine derivative cerivastatin is a new entirely synthetic and enantiomerically pure inhibitor of 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase. As a sodium salt cerivastatin is present in the active, open ring form. Cerivastatin inhibited the membrane-bound (non-solubilized) HMG-CoA reductase of the native microsomal fraction isolated from rat liver with a Ki value of 1.3 x 10(-9) M. The reference compound lovastatin was 100-fold less potent and exhibited a Ki value of 150 x 10(-9) M. Cerivastatin inhibited the cholesterol synthesis in the human hepatoma cell line HepG2 cells with a similar IC50 value of 1.0 x 10(-9) M. In vivo studies reflected its high in vitro activity. In both rats and dogs, cerivastatin inhibited the hepatic [14C]cholesterol synthesis from [14C]acetate with an oral ED50 value of 0.002 mg/kg body weight, while lovastatin exhibited an oral ED50 value of 0.3 mg/kg in rats, showing again the ratio of 100 or more between cerivastatin and lovastatin. In the small intestine and testes, cerivastatin was at least 50-fold less active with oral ED50 values higher than 0.1 mg/kg, which is indicative for a high liver selectivity of cerivastatin. In cholestyramine-primed dogs cerivastatin dose dependently lowered the serum cholesterol concentrations by up to 59% with 0.1 mg/kg after 20 days. Interestingly, the serum triglycerides were markedly reduced by 53 and 76% with 0.03 and 0.1 mg/kg, respectively. In normal chow fed dogs the low density lipoprotein (LDL) concentrations were reduced by up to 75% after 0.1 mg cerivastatin/kg. The ratio of HDL/LDL increased by 81% compared with a change of only 14% in the placebo treated control group. The antiatherogenic effect of cerivastatin was shown in rabbits fed a diet enriched with 0.2% cholesterol. After 9 weeks on diet 0.1 mg cerivastatin/kg decreased the accumulation of cholesterol ester in the arterial tissue by 73%. In summary, these data as compared to published data on other HMG-CoA reductase inhibitors demonstrate cerivastatin to be the most active compound in this class. Vastatins used in therapy are effective in mg doses, while cerivastatin offers a new low dose therapy in the microg range. PMID- 9395279 TI - Cardiovascular prognosis in relation to apolipoproteins and other lipid parameters in patients with stable angina pectoris treated with verapamil or metoprolol: results from the Angina Prognosis Study in Stockholm (APSIS). AB - Relationships between apolipoproteins and other lipid parameters and cardiovascular (CV) prognosis were evaluated in the Angina Prognosis Study In Stockholm (APSIS). Out of 809 patients with stable angina pectoris, lipid variables were obtained in 786 patients at baseline, and after one month's double blind treatment with metoprolol or verapamil, to evaluate treatment effects on these lipid variables. During a median follow-up time of 3.3 years (2663 patient years), 37 patients suffered a CV death, 30 suffered a non-fatal myocardial infarction (MI) and 100 underwent a revascularization. Apolipoprotein (apo) A-I, high-density lipoprotein cholesterol and triglycerides were predictors of CV death or non-fatal MI in univariate analyses, but only apo A-I remained as an independent predictor in multivariate analyses. All lipid variables except low density lipoprotein cholesterol were related to the risk of revascularization in univariate analyses, but only apo A-I and apo B were independent predictors of such events. Triglycerides were weakly, but not independently, associated with prognosis. Verapamil and metoprolol had differential short-term effects on lipids, with a shift towards a more atherogenic profile in metoprolol treated patients. However, there was no significant impact of the treatment given, or of these treatment effects on the risk of CV events. Results of the present study suggest that apolipoprotein levels were better predictors of CV events than other lipid parameters in patients with stable angina pectoris. PMID- 9395281 TI - Ras-like GTPases. PMID- 9395282 TI - Tales of tails: Brachyury and the T-box genes. PMID- 9395283 TI - Regulation and function of the JNK subgroup of MAP kinases. PMID- 9395284 TI - TGF-beta receptor signaling. PMID- 9395285 TI - The role of the bcl-2/ced-9 gene family in cancer and general implications of defects in cell death control for tumourigenesis and resistance to chemotherapy. AB - Cell production within an organ is determined by the rate of immigration, proliferation, differentiation, emigration and death of cells. Abnormalities in any one of these processes will disturb normal control of cell production, thereby eliciting hyperplasia can be an early event in neoplasia. Cell death, apoptosis, is a physiological process responsible for removing unwanted cells. It is used in multi-cellular organisms for tissue remodelling during embryogenesis, regulation of cell turnover and as a defence strategy against invading pathogens. In this review article we describe the role of the bcl-2/ced-9 gene family in cancer and discuss the general implications of defects in the apoptosis program for tumourigenesis and resistance of cancer cells to chemotherapy in light of current knowledge of the molecular mechanisms of cell death. PMID- 9395286 TI - Functional role for the c-Abl protein tyrosine kinase in the cellular response to genotoxic stress. PMID- 9395287 TI - Targeted therapy of cancer with recombinant immunotoxins. PMID- 9395288 TI - Alzheimer's alpha-secretase may be a calcium-dependent protease. AB - Proteolytic processing of beta-amyloid precursor protein (APP) is believed to be fundamental to the understanding of Alzheimer's disease. The identities and the regulatory elements of the proteases involved in the process, known as alpha/beta/gamma secretases, are unclear. In this study, by examining reported data, we found some indications suggesting that the putative alpha-secretase may be a calcium-dependent protease, and that this enzyme may play a primary role in the regulation of APP processing. Based on this, we proposed a model for the membrane orientations of the secretases for further discussions. PMID- 9395289 TI - Purified HrpA of Pseudomonas syringae pv. tomato DC3000 reassembles into pili. AB - Pseudomonas syringae pv. tomato DC3000 produces Hrp pili under inducing in vitro conditions. A preparation of partially purified extracellular filaments contains HrpA, flagellin and some minor contaminants. HrpA was separated from the major contaminant, the flagellin, by gel filtration to a fraction containing HrpA as well as its three N-terminally truncated forms. These were further separated by two steps of reversed phase chromatography. HrpA and its degradation products were each shown to reassemble into filament structures after denaturation and renaturation showing that HrpA alone is sufficient for formation of filament structures. PMID- 9395290 TI - Activation of sphingosine kinase in pheochromocytoma PC12 neuronal cells in response to trophic factors. AB - Nerve growth factor (NGF), basic fibroblast growth factor (bFGF), dibutyryl cAMP and forskolin, known differentiating agents for pheochromocytoma PC12 cells, induced sustained activation of sphingosine kinase, the enzyme responsible for the formation of the sphingolipid second messenger, sphingosine-1-phosphate, which mediates the mitogenic effects of certain growth factors. In contrast, epidermal growth factor and insulin-like growth factor-1, which stimulate proliferation of PC12 cells, induced only small and transient increases in sphingosine kinase activity. Of the growth factors examined, NGF was the most potent activator of sphingosine kinase, inducing a 4-fold increase in Vmax. Sphingosine kinase activity induced by NGF, but not FGF, was blocked by the protein kinase inhibitor K252a when added simultaneously, with minimal effect when added after 60 min. Thus, activation of sphingosine kinase may have an important role in neural differentiation. PMID- 9395291 TI - Cloning of a functional vesicular GABA and glycine transporter by screening of genome databases. AB - The unc-47 locus of Caenorhabditis elegans has been suggested to encode a synaptic vesicle GABA transporter. Here we used hydropathy plot analysis to identify a candidate vesicular GABA transporter in genomic sequences derived from a region of the physical map comprising unc-47. A mouse homologue was identified and cloned from EST database information. In situ hybridization in rat brain revealed codistribution with both GABAergic and glycinergic neuronal markers. Moreover, expression in COS-7 and PC12 cells induced an intracellular, glycine sensitive GABA uptake activity. These observations, consistent with previous data on GABA and glycine uptake by synaptic vesicles, demonstrate that the mouse clone encodes a vesicular inhibitory amino acid transporter. PMID- 9395292 TI - Early expression of a beta1-adrenergic receptor and catecholamines in Xenopus oocytes and embryos. AB - From a Xenopus stage 11 cDNA library, we have cloned a gene, termed X-beta1AR, whose sequence is highly homologous to that of the human beta1-adrenergic receptor. As shown by RT-PCR assay, X-beta1AR RNA is present in the mature oocyte, decreases after fertilization up to stage 6 and then gradually increases during gastrulation. Binding studies performed with radiolabeled ligands reveal that X-beta1AR RNA is translated into the receptor protein. Furthermore, noradrenaline and adrenaline are also detected in oocytes and early embryos. The concomitant presence of beta1-adrenergic receptors and catecholamines suggest that this ligand-receptor couple could play a role in the very early stages of embryonic development. PMID- 9395293 TI - C-terminally deleted fragments of 40-kDa earthworm actin modulator still show gelsolin activities. AB - C- and N-terminally truncated fragments of earthworm gelsolin were constructed, cloned and expressed in Escherichia coli. G-actin-binding properties of these fragments and their influences on the polymeric state of actin were investigated. A construct lacking a large part of the third segment [E(1-295)] supports actin nucleation similar to the complete protein and shows reduced actin fragmentation property, but is no longer Ca2+-sensitive in its activity. The first and the second segments (E1 and E2) each contain one actin-binding site. In contrast to human gelsolin, E1 in combination with a short N-terminal region of E2 is not sufficient for the F-actin-severing activity of the protein. PMID- 9395294 TI - Inhibitory effect on curcumin on mammalian phospholipase D activity. AB - Curcumin, the major yellow pigment of turmeric (Curcuma longa), has strong anti carcinogenic and anti-inflammatory activities. We examined the effects of curcumin on enzyme activities of the following phospholipases in a cell-free system: G protein-mediated phospholipase D (PLD), phosphatidylinositol-specific phospholipase C, and phospholipase A2 from mouse macrophage-like cell line J774.1 cells, sphingomyelinase from bovine brain, and phosphatidylcholine-phospholipase C from Bacillus cereus. Curcumin inhibited several types of phospholipases, most effectively PLD among those tested. It also inhibited 12-O-tetradecanoylphorbol 13-acetate-induced PLD activation in intact J774.1 cells in a dose-dependent manner. These results suggest that the anti-inflammatory and anti-carcinogenic action of curcumin is partly due to the inhibition of PLD. PMID- 9395295 TI - Cationic lipids destabilize lysosomal membrane in vitro. AB - Addition of cationic lipids to plasmid DNA considerably increases the efficiency of transfection. The mechanism has not yet been elucidated. A possibility is that these compounds destabilize biological membranes (plasma, endosomal, lysosomal), facilitating the transfer of nucleic molecules through these membranes. We have investigated the problem by determining if a cationic lipid N-(1-(2,3 dioleoxy)propyl)-N,N,N,-trimethylammonium methyl-sulfate (DOTAP, Boehringer, Mannheim, Germany) affects the integrity of rat liver lysosomal membrane. We have measured the latency of beta-galactosidase, a lysosomal enzyme, and found that incubation of lysosomes with low concentrations of DOTAP causes a striking increase in free activity of the hydrolase and even a release of the enzyme into the medium. This indicates that lysosomal membrane is deeply destabilized by the lipid. The phenomenon depends on pH, it is less pronounced at pH 5 than at pH 7.4. Anionic compounds, particularly anionic amphipathic lipids, can to some extent prevent this phenomenon. It can be observed with various cationic lipids. A possible explanation is that cationic liposomes interact with anionic lipids of lysosomal membrane, allowing a fusion between the lipid bilayers which results in a destabilization of the organelle membrane. PMID- 9395296 TI - Degradation of the 74 kDa form of L-histidine decarboxylase via the ubiquitin proteasome pathway in a rat basophilic/mast cell line (RBL-2H3). AB - L-Histidine decarboxylase (HDC) is a dimer consisting of two identical 53 kDa subunits. On the other hand, the size of HDC deduced from its cDNA sequence is around 74 kDa, indicating that the translated 74 kDa form of HDC is subjected to post-translational processing to generate the 53 kDa form. However, modification of the translated 74 kDa form of HDC in histamine-forming cells is unknown. Here we demonstrate that the 74 kDa form is translated in rat basophilic leukemia cells, followed by conversion to the 53 kDa form, and that the 74 kDa form is a short half-life protein because of the degradation mediated by the ubiquitin proteasome pathway. Degradation of the 74 kDa form was stimulated in the presence of an ATP-generating system, accompanied by ubiquitination, and inhibited by specific proteasome inhibitors such as ZL3H and lactacystin. A significant amount of proteasome activity was detected in RBL-2H3 cells. PMID- 9395297 TI - Expression of B-type endothelin receptor gene during neural development. AB - Mutations of the B-type endothelin receptor (ETRB) gene have been found to cause defects in the development of enteric neurons, which resulted in aganglionic megacolon in rodents and humans. To determine the distribution of ETRB mRNA during neural development, mainly in the CNS, in situ hybridization was applied at various developmental stages of rat. ETRB gene was abundantly expressed prenatally in the ventricular and subventricular zones, as well as postnatally in the ependymal and subependymal cells. ETRB mRNA was also strongly detected prenatally in the dorsal root ganglia, as well as postnatally in the cerebellar Bergmann glial cells and epithelial cells of choroid plexus. Our data suggest that ETRB acts as a regulator in the differentiation, proliferation, or migration of neural cells during development. PMID- 9395298 TI - Characterization of nuclear tRNA(Tyr) introns: their evolution from red algae to higher plants. AB - We have previously isolated numerous intron-containing nuclear tRNA(Tyr) genes derived from either monocotyledonous (Triticum) or dicotyledonous (Arabidopsis, Nicotiana) plants by screening the corresponding genomic phage libraries with a synthetic tRNA(Tyr)-specific oligonucleotide. Here we have characterized additional tRNA(Tyr) genes from phylogenetically divergent plant species representing red algae (Champia), brown algae (Cystophyllum), green algae (Ulva), stonewort (Chara), liverwort (Marchantia), moss (Polytrichum), fern (Rumohra) and gymnosperms (Ginkgo) using amplification of the coding sequences from the corresponding genomic DNAs by polymerase chain reaction (PCR). All novel tRNA(Tyr) genes contain intervening sequences of variable sequence and length ranging in size from 11 to 21 bp. However, two features are conserved in all plant pre-tRNA(Tyr) introns: they possess a uridine and less frequently an adenosine at the 5' boundary and can adopt similar intron secondary structures in which an extended anticodon helix of 4-5 bp is formed by base-pairing between nucleotides of the intron and the anticodon loop. In order to elucidate the potential role of the highly conserved uridine at the first intron position, we have replaced it by all other nucleosides in an Arabidopsis pre-tRNA(Tyr) and have studied in wheat germ extract its effect on splicing and on conversion of U to psi in the GpsiA anticodon. Furthermore, we discuss the putative acquisition of tRNA(Tyr) introns at an early step of evolution after the separation of Archaea and Eucarya. PMID- 9395299 TI - Mutants affected in the putative diacylglycerol binding site of yeast protein kinase C. AB - In an attempt to study the functional similarities between protein kinase C from the yeast Saccharomyces cerevisiae and its human homologues we have started in vitro mutagenesis to alter specific domains. Here we report on the exchange of four cysteine residues by serines in yeast Pkc1p that have been shown to be essential for diacylglycerol (DAG) binding and activation by this compound in humans. The mutant yeast protein leads to sensitivity to caffeine and low concentrations of SDS when expressed in a pkc1 deletion strain. However, sensitivity to staurosporine was not affected. Our data indicate that the conserved DAG binding domain serves an important function in yeast Pkc1p. PMID- 9395300 TI - A stable intermediate product of the archaeal zinc-containing 7Fe ferredoxin from Sulfolobus sp. strain 7 by artificial oxidative conversion. AB - Irreversible conversion of the purified zinc-containing 7Fe ferredoxin from the thermoacidophilic archaeon Sulfolobus sp. strain 7 was found to occur under aerobic conditions at pH 5.0 and at 4 degrees C. This process accompanied a substantial increase of the electron paramagnetic resonance signal attributed to a [3Fe-4S]1+ cluster, and the converted form containing approximately 6 Fe/Zn (mol/mol) had a net charge different from that of the native 7Fe ferredoxin. These data provide evidence for the formation of a stable intermediate product of the archaeal ferredoxin having two [3Fe-4S] clusters and a zinc center by artificial oxidative degradation. This further explains the discrepancy that a zinc center and two [3Fe-4S] clusters (but not a zinc center and one [3Fe-4S] cluster plus one [4Fe-4S] cluster) are observed in the crystal structure at pH 5.0. PMID- 9395301 TI - Glycosylation of RNA polymerase II from wheat germ. AB - RNA polymerase II from wheat germ was analyzed for the presence of sugars. The two largest subunits and the 27 and 25 kDa subunits were found to be glycosylated by a variety of sugars. However, no N-acetylglucosamine was detected, which was found by Kelly et al. (J. Biol. Chem. (1993) 268, 10416-10424) in the largest subunit of RNA polymerase II from calf thymus. Thus it appears that the regulatory function of this sugar, postulated by Kelly et al., is performed in the wheat germ enzyme by other monosaccharides. Carbohydrate analysis of the two largest subunits of the calf thymus enzyme also revealed the presence, beside N acetylglucosamine, of other sugars. Some similarities in the features of glycosylation of the two polymerases, isolated from very different organisms, suggest that the sugar moieties have an important role in the structure and/or function of these enzymes. PMID- 9395302 TI - Activated human neutrophils rapidly break down nitric oxide. AB - Isolated human neutrophils produced no detectable (< 10 nM) nitric oxide (NO) before or after activation with phorbol 12-myristate 13-acetate (PMA) or a chemotactic peptide, N-formyl-L-methionyl-L-leucyl-L-phenylalanine. Physiological levels of NO (1 microM) added before or after neutrophil activation had no effect on their respiratory burst oxygen consumption. Neutrophils activated with PMA caused very rapid breakdown of exogenously added NO. NO breakdown rates recorded at 250 nM NO were 0.09 +/- 0.02 and 3.77 +/- 0.23 nmol NO/min/10(6) cells (n = 3) before and after activation respectively and addition of copper-zinc superoxide dismutase during activation significantly decreased this rate (1.06 +/- 0.09 nmol NO/min/10(6) cells (n = 3)), suggesting that superoxide (O2-) production was mainly responsible for the NO breakdown. These results suggest that activation of human neutrophils in vivo will dramatically decrease surrounding NO levels, potentially causing vasoconstriction, platelet aggregation and adhesion and peroxynitrite (ONOO-) formation. PMID- 9395303 TI - Insulin-like growth factor I reverses interleukin-1beta inhibition of insulin secretion, induction of nitric oxide synthase and cytokine-mediated apoptosis in rat islets of Langerhans. AB - We have previously observed that treatment of rat islets of Langerhans with interleukin-1beta for 12 h results in nitric oxide-dependent inhibition of insulin secretion, while 48 h treatment increased rates of islet cell death by apoptosis. Here, we demonstrate that interleukin-1beta-mediated nitric oxide formation and inhibition of insulin secretion are significantly reduced by 24 h pretreatment of rat islets of Langerhans with insulin-like growth factor I (IGF I). IGF-I decreased cytokine induction of nitric oxide synthase in islets. Use of an arginine analogue in culture or IGF-I pretreatment of islets were also effective in protecting islets against cytokine-mediated apoptotic cell death. We conclude that IGF-I antagonises inhibitory and cytotoxic effects of cytokines in rat islets. PMID- 9395304 TI - Folding of the Fab fragment within the intact antibody. AB - At present it is not clear to which extent the Fab fragment and the Fc part of an antibody interact in the intact immunoglobulin structure. To determine such potential interactions the unfolding and refolding of an isolated Fab fragment and the respective antibody MAK 33 (kappa/IgG1) are compared. It could be shown that the proline independent renaturation kinetics of both an unfolding intermediate and the fully denatured form of both proteins are identical. Upon denaturation, the loss of antigen binding activity occurs with the same rate for both the Fab fragment and the intact antibody. However, the complete structural unfolding of the Fab part of the antibody is significantly slower than that of the isolated Fab fragment. These kinetic data suggest that the structure of the Fab fragment within the intact antibody is stabilized by interactions, presumably with the Fc part, missing in the isolated Fab. PMID- 9395305 TI - Binding of multiple ligands to pleckstrin homology domain regulates membrane translocation and enzyme activity of beta-adrenergic receptor kinase. AB - Pleckstrin homology (PH) domains are discrete structural modules present in numerous proteins involved in signal transduction processes. In the case of the beta-adrenergic receptor kinase (betaARK), PH domain-mediated binding of two ligands, the betagamma subunits of heterotrimeric G proteins (Gbetagamma) and phosphatidylinositol 4,5-bisphosphate (PIP2), has been shown to be required for the kinase function. In this study, the ability of Gbetagamma and PIP2 to affect membrane localization of betaARK is used to define the ligand binding characteristics of the betaARK PH domain. The binding of these ligands to the PH domain of the intact kinase is shown to be cooperative, Gbetagamma increasing the affinity of the PH domain for PIP2. Notably, although PIP2-dependent membrane association of betaARK is observed at high concentrations of this lipid, in the absence of Gbetagamma, no receptor phosphorylation is observed. Peptides derived from the receptor intracellular loop inhibit the receptor phosphorylation without affecting the membrane translocation of the kinase complex, suggesting that betaARK activity does not necessarily correlate with the amount of betaARK associated with the membrane. These results point to a distinct role for each PH domain ligand in betaARK-mediated receptor phosphorylation. Strikingly, the ligand binding characteristics of the isolated betaARK PH domain fused to glutathione S-transferase are very different from those of the PH domain of the intact kinase. Thus, in contrast to the native protein, the isolated PH domain binds Gbetagamma and PIP2 independently and with no apparent cooperativity. That protein environment plays an important role in determining the ligand binding characteristics of a particular PH domain highlights the potential risks of inferring mechanisms from studies of isolated PH domains. PMID- 9395306 TI - Hemin and related porphyrins inhibit beta-amyloid aggregation. AB - Porphyrins related to the naturally occurring pigment heme were found to effectively interfere with the aggregation of beta-amyloid peptides as determined by an immunoassay configured for the detection of beta-amyloid oligomers. Oligomerisation of beta-amyloid is believed to be a key event in the progression of Alzheimer's disease. Inhibition of this aggregation is thus an important strategy in combating this commonest form of senile dementia. Evidence was also generated for hemin and hematin mediated protection of cultured cells against the neurotoxic effects of beta-amyloid. These data are discussed with reference to the known pathology of Alzheimer's disease and the chemistry of porphyrins. PMID- 9395307 TI - Identification of potential ferric binding residues in the iron-binding protein of pathogenic Neisseria meningitidis through structure-based multiple sequence alignments. AB - The ferric iron-binding proteins of pathogenic Neisseria display structural and metal-binding properties characteristic of the transferrin family. In the absence of structural data for the ferric iron-binding proteins, spacial folding templates have been derived for the meningococcal protein from complete and partial structure-based multiple sequence alignments with structurally related proteins. The templates have been used to identify a number of potential iron binding residues. These include four residues that are identical with the iron coordinating ligands of transferrin, but only two reside within equivalent structural elements. PMID- 9395308 TI - The role of tyrosine phosphorylation in proliferation and maturation of erythroid progenitor cells--signals emanating from the erythropoietin receptor. AB - Red blood cells arise continuously from pluripotent stem cells which mature and become functionally specialized upon commitment to the erythroid lineage. In mammals, the key regulator of this process is the hormone erythropoietin (EPO). Hormone binding to the cognate receptor, the erythropoietin receptor (EPO-R), causes receptor homodimerization and transiently triggers tyrosine phosphorylation within target cells. Although the EPO-R lacks intrinsic enzymatic activity it couples, presumably sequentially, to the protein tyrosine kinase receptor c-KIT and the cytosolic protein tyrosine kinase JAK2. Signaling through the EPO-R is promoted by tyrosine phosphorylation of the cytosolic domain and the recruitment of secondary signaling molecules such as the lipid kinase inositolphospholipid 3-kinase (phosphatidylinositol 3-kinase) and protein tyrosine phosphatase SHP-2 to the activated receptor. Complex formation of the activated EPO-R with the protein tyrosine phosphatase SHP-1 terminates signaling. In primary fetal liver cells redundant signals emanating from phosphotyrosine residues in the EPO-R support formation of erythroid colonies in vitro. However, since the last tyrosine residue in the cytosolic domain of the EPO-R, Y479, uniquely supports in the absence of other tyrosine residues an almost normal level of colony-forming unit-erythroid (CFU-E) colony formation, Y479 represents one of the key residues required in vivo for erythroid proliferation and differentiation. The signal emanating from Y479 involves sequential EPO-induced recruitment of phosphoinositol lipid 3-kinase to the EPO-R and activation of mitogen-activated-protein(MAP)kinase activity. The MAP-kinase signaling cascade could serve as an intracellular switch integrating signals mediated by several phosphotyrosine residues in the cytosolic domain of the EPO-R and provide a possible explanation for partial redundancy in signaling. PMID- 9395309 TI - Signal transduction in fibroblasts stably transformed by [Val12]Ras--the activities of extracellular-signal-regulated kinase and Jun N-terminal kinase are only moderately increased, and the activity of the delta-inhibitor of c-Jun is not alleviated. AB - Ras-transformed cells often show high levels of expression of activating protein 1 and Ets and of genes regulated by these transcription factors. In analogy with the effects of transient stimulation of Ras, it is assumed that the increase in transcription-factor transactivation in stably transformed cells is due to Ras induced constitutive activation of mitogen-activated protein kinases. However, this has not been extensively studied. Using specific substrate peptides, we have examined here the activities of two types of mitogen-activated protein kinase, extracellular-signal-regulated kinase (ERK) and Jun N-terminal kinase (JNK), in [Val12]Ras-transformed rat embryo fibroblast cell lines. These activities were elevated 2-3-fold in Ras-transformed cells compared with non-transformed cells with a similar growth rate. Increased ERK activity was not necessarily accompanied by a similar increase in JNK activity. In transformed cells, ERK and JNK activities could be stimulated fourfold and ninefold by phorbol ester and ultraviolet-light treatment, respectively, indicating that only a fraction of these enzymes were constitutively activated in these cells. It has been suggested that inactive JNK downregulates c-Jun transcriptional activity by binding to the c-Jun delta-domain. No decrease in delta-inhibitor activity could be demonstrated in Ras-transformed cells compared with control cells, consistent with the presence of mainly inactive JNK in transformed cells. Treatment of transformed cells wih benzodiazepine 5B, an inhibitor of Ras farnesylation, decreased ERK and JNK activities, and concomitantly caused morphological reversion, reduced growth rate, and normalization of transformation-related gene expression. We conclude that in stably Ras-transformed cells the moderately increased ERK/JNK activities are not coregulated, and that ERK rather than JNK activity correlated with transformation-related gene expression. PMID- 9395310 TI - Transport of activated fatty acids by the peroxisomal ATP-binding-cassette transporter Pxa2 in a semi-intact yeast cell system. AB - In the yeast Saccharomyces cerevisiae, fatty acid beta-oxidation is restricted to peroxisomes. Previous studies have shown two possible routes by which fatty acids enter the peroxisome. The first route involves transport of medium-chain fatty acids across the peroxisomal membrane as free fatty acids, followed by activation within the peroxisome by Faa2p, an acyl-CoA synthetase. The second route involves transport of long-chain fatty acids. Long-chain fatty acids enter the peroxisome via a route that involves activation in the extraperoxisomal space, followed by transport across the peroxisomal membrane. It has been suggested that this transport is dependent upon the peroxisomal ATP-binding-cassette transporters Pxa1p and Pxa2p. In this paper we investigated whether Pxa2p is directly responsible for the transport of C18:1-CoA, a long-chain acyl-CoA ester. Using protoplasts in which the plasma membrane has been selectively permeabilised by digitonin, we show that C18:1-CoA, but not C8:0-CoA, enters the peroxisome via Pxa2p, in an ATP-dependent fashion. The results obtained may contribute to the elucidation of the primary defect in the human disease X-linked adrenoleukodystrophy. PMID- 9395311 TI - Comparative stability and clearance of [Met30]transthyretin and [Met119]transthyretin. AB - [Met119]Transthyretin has been described as a non-amyloidogenic transthyretin variant. In Portugal, it has also been found in compound heterozygotic individual carriers of [Met30]transthyretin, the most prevalent variant associated with familial amyloidotic polyneuropathy. In these individuals, the evolution of the disease seems to be more benign than in typical [Met30]transthyretin carriers, suggesting a protective effect of [Met119]transthyretin on the pathogenic effects of [Met30]transthyretin. To study the mechanisms of this protective effect, we performed comparative in vivo clearance studies. Heterotetrameric [Met119]transthyretin showed a slower clearance, whereas homotetrameric [Met30]transthyretin presented a faster clearance. These data correlate with the relative TTR levels present in carriers of these mutations. Comparative analyses of the resistance to dissociation into monomers of serum transthyretin by 4M urea isoelectric focusing suggested a higher tetrameric stability of transthyretin in [Met119]transthyretin carriers, in contrast to a lower stability in [Met30]transthyretin carriers. The compound heterozygotes presented a pattern similar to the normal individuals. Our results suggest that the protective clinical effect of the Met119 mutation possibly involves the stabilisation of the tetrameric structure of transthyretin. Whether this behaviour correlates with the different metabolism found for the two variants is not known. The approaches reported here open some possibilities for the study and development of future therapeutic agents of familial amyloidotic polyneuropathy. PMID- 9395312 TI - Regulation of argininosuccinate synthetase mRNA level in rat foetal hepatocytes. AB - Expression of the hepatic gene for argininosuccinate synthase (ASS), one of the key enzymes of the urea cycle, was analysed during the perinatal period in the rat. To this end, the amount of specific mRNA was measured in the liver at various stages of development and in cultured foetal hepatocytes maintained in different hormonal conditions. The ASS mRNA was first detected in 15.5-day foetuses and its level increased concomitantly with a rise in the enzyme activity, suggesting that the appearance of the ASS activity reflects the turning on of specific gene transcription. This was demonstrated by run-on assay which showed an enhanced rate of transcription of the ASS gene during the perinatal period. When foetal hepatocytes were cultured with dexamethasone, a dose dependent increase in ASS mRNA was measured, which was completely abolished by actinomycin D addition. The transcription rate of the gene was increased about twofold in the presence of the steroid, as measured by nuclear run-on assay. This transcriptional action could additionally require a protein factor since it could be inhibited by the simultaneous addition of puromycin. Insulin or glucagon respectively repressed or enhanced the dexamethasone-induced accumulation of ASS mRNA when added simultaneously with the steroid for 24 h. This developmental regulation of the ASS mRNA by glucocorticoids, insulin and glucagon could account for the modulation of the enzyme activity previously observed in vivo and in vitro in the foetal liver. PMID- 9395313 TI - 5' sequences direct developmental expression and hormone responsiveness of tyrosine aminotransferase in primary cultures of fetal rat hepatocytes. AB - Tyrosine aminotransferase (TyrAT) is one of several gluconeogenic enzymes which appear postnatally in humans and rodents in response to increased glucocorticoid and glucagon levels and decreased insulin. Primary cultured fetal rat hepatocytes older than day 15 of gestation (>E15) transcribe the TyrAT gene in response to the synergistic effect of dexamethasone and N6,2'-O-dibutyryl-adenosine 3',5' monophosphate (Bt2cAMP), whereas less mature hepatocytes (E15 hepatocytes, and not 3)-alpha-D-ManpNAc (1-->4)-beta-D-GlcpA-(1-->3)-beta-D-Ga lp-(1-->3)-beta-D-GlcpNAc-(1-->. PMID- 9395324 TI - The role of subunit VIII in the structural stability of the bc1 complex from Saccharomyces cerevisiae studied using hybrid complexes. AB - The QCR8 genes encoding subunit VIII of the bc1 complex from Kluyveromyces lactis and Schizosaccharomyces pombe partially complement the respiratory-deficient phenotype of a S. cerevisiae QCR8-null mutant. This implies that the heterologous Qcr8 subunits can be imported by S. cerevisiae mitochondria and that they assemble to form a hybrid bc1 complex that is sufficiently active to support growth. In contrast, the QCR8 gene from bovine heart, encoding the 9.5-kDa subunit, is not able to restore respiratory function to the S. cerevisiae null mutant. This lack of functional complementation is directly attributable to the inability of S. cerevisiae mitochondria to import this protein as shown by in vitro assays. However, a hybrid gene encoding the N-terminal 26 residues of S. cerevisiae subunit VIII and the rest of the 9.5-kDa bovine heart homologue, was able to functionally complement the QCR8-null mutant, albeit to a very low extent. Successful import into S. cerevisiae mitochondria was confirmed by in vitro import experiments. Surprisingly, although assembly of these hybrid complexes is reduced to an extent that is proportional to the evolutionary distance of the homologue to S. cerevisiae, the specific activities of the assembled complexes are the same as for the wild-type bc1 complex. After solubilisation of the mitochondrial membranes with the mild detergent dodecyl maltoside, the wild-type enzyme can be inactivated by incubation at increased temperature, independent of protease activity. The rate of inactivation can be significantly increased by the addition of o-phenanthroline [Boumans, H., Grivell, L. A. & Berden, J. A. (1997) J. Biol. Chem. 272, 16753-16760]. The hybrid complexes are much more sensitive to both types of treatment. We conclude that substitution of subunit VIII by a homologous counterpart results in a loosening of the structure of the bc1 complex on the intermembrane space side, resulting in a less stable insertion of the Rieske Fe-S protein in vivo and therefore a lower stability of the assembled enzyme under certain in vitro conditions, but without an effect on catalytic activity. PMID- 9395325 TI - The Na+-translocating NADH:ubiquinone oxidoreductase from Vibrio alginolyticus- redox states of the FAD prosthetic group and mechanism of Ag+ inhibition. AB - The FAD prosthetic group of the Na+-motive NADH:ubiquinone oxidoreductase (Na+ NQR) from Vibrio alginolyticus was investigated by ultraviolet-visible and fluorescence spectroscopy. The reduction of Na+-NQR by excess NADH in the presence of 6-13 microM O2 resulted in the formation of the blue flavosemiquinone radical. If the concentration of dioxygen was further reduced to 0.1 microM O2, neither the reduction of Na+-NQR by NADH nor its reoxidation with ubiquinone-1 (Q 1) yielded a stable flavosemiquinone in equilibrium with reductant or oxidant, respectively, but the fully reduced (Fl(red)H2) or oxidized flavin (Fl(ox)) prevailed. During reoxidation of Fl(red)H2 with Q-1, the intermediate formation of an absorbance band around 800 nm was observed, which was tentatively assigned as the Fl(red)H(-)-NAD+ charge-transfer complex. Complete reoxidation of Fl(red)H2 in Na+-NQR was achieved by a fivefold excess of Q-1 over NADH. These results indicated that only a small fraction of FAD was in the flavosemiquinone redox state during turnover to mediate the electron transfer between the hydride donor, NADH, and the one-electron acceptor [2Fe-2S]. The titration of Na+-NQR with Ag+, a specific inhibitor, was followed by the fluorescence emission spectra of FAD (Fl(ox)). The addition of Ag+ resulted in a marked increase of the flavin fluorescence (16% at 200 nM Ag+), with half-maximal saturation at approximately 50 nM Ag+, indicating dissociation of FAD from the enzyme. The increase in fluorescence intensity correlated with the loss of enzyme activity. Gel filtration of the Ag+-treated Na+-NQR confirmed that FAD had been displaced from the holo-enzyme. PMID- 9395326 TI - Flavin adenine dinucleotide and flavin mononucleotide metabolism in rat liver- the occurrence of FAD pyrophosphatase and FMN phosphohydrolase in isolated mitochondria. AB - In order to gain some insight into mitochondrial flavin biochemistry, rat liver mitochondria essentially free of lysosomal and microsomal contamination were prepared and their capability to metabolise externally added and endogenous FAD and FMN tested both spectroscopically and via HPLC. The existence of two novel mitochondrial enzymes, namely FAD pyrophosphatase (EC 3.6.1.18) and FMN phosphohydrolase (EC 3.1.3.2), which catalyse FAD-->FMN and FMN-->riboflavin conversion, respectively, is shown. They differ from each other and from extramitochondrial enzymes, as judged by their pH profile and inhibitor sensitivity, and can be separated in a partial FAD pyrophosphatase purification. Digitonin titration and subfractionation experiments show that FAD pyrophosphatase is located in the outer mitochondrial membrane and FMN phosphohydrolase in the intermembrane space. Since these enzymes can metabolise endogenous FAD and FMN, which are made available by using both Triton X-100 and the effector oxaloacetate, a proposal is made that FAD pyrophosphatase and FMN phosphohydrolase play a major role in mitochondrial flavoprotein turnover. PMID- 9395327 TI - Purification and characterization of tilapia (Oreochromis mossambicus) deoxyribonuclease I--primary structure and cDNA sequence. AB - DNase I of tilapia (Oreochromis mossambicus) was purified to homogeneity. Tilapia DNase I is most active at pH 8.5 with Mg2+ as activator. The Ca2+/Mg2+ pair has a synergistic effect on activation. The enzyme is readily inactivated by heating above 55 degrees C, but is not inactivated by trypsin or 2-mercaptoethanol under alkaline conditions, with or without CaCl2. Its isoelectric point is 6.0. The 258 amino-acid sequence of tilapia DNase I was derived from overlapping sequences of tryptic, chymotryptic and CNBr peptides. The purified enzyme has two variants differing by a single Lys-->Arg mutation at position 125. The polypeptide chain has one disulfide bridge and one carbohydrate side chain. By mass spectrometry, the purified enzyme shows many molecular mass forms differing by Lys/Arg substitution and sugar-chain length. The major form has a molecular mass of 30,914 Da. A 1061-bp nucleotide sequence for the cDNA of tilapia DNase I, obtained by gene cloning and DNA sequencing, contains an ORF coding for a putative 26-residue transmembrane peptide and the mature DNase I polypeptide. PMID- 9395328 TI - Interleukin-3 activates Syk in a human myeloblastic leukemia cell line, AML193. AB - Protein-tyrosine kinases and phosphatases play an important role in cytokine mediated cell growth. The proliferation of a human myeloid leukemia cell line, AML193, is dependent on interleukin-3 (IL-3) or granulocyte colony-stimulating factor. In the current study, we demonstrated that a non-receptor-type protein tyrosine kinase, Syk, was immediately activated by the stimulation with IL-3 or granulocyte colony-stimulating factor in AML193 cells. We further investigated the relation of Syk with IL-3-mediated signaling and found that the IL-3 receptor beta subunit was immunoprecipitated with Syk. Since the IL-3 receptor beta subunit is known to mediate growth signaling, our results indicate that Syk may be involved in the proliferation of myeloid cells. PMID- 9395329 TI - Change in the mode of gene expression of the hypopharyngeal gland cells with an age-dependent role change of the worker honeybee Apis mellifera L. AB - Major proteins synthesized in the hypopharyngeal gland of the worker honeybee change from bee-milk proteins to alpha-glucosidase in accordance with the age dependent role change of the worker bee. Previously, we showed that the gene for alpha-glucosidase is expressed specifically in the forager-bee gland [Ohashi, K., Sawata, M., Takeuchi, H., Natori, S. & Kubo, T. (1996) Biochem. Biophys. Res. Commun. 221, 380-385]. Here, we describe the isolation and analysis of cDNAs for two bee-milk 56-kDa and 64-kDa proteins. The 56-kDa protein was a glycoprotein which shared 63.2% and 56.9% amino acid sequence identities with proteins encoded by cDNA for royal-jelly-related protein 57-1 (pRJP57-1) and pRJP57-2. The 64-kDa protein cDNA was identical to pRJP57-1. Thus, these bee-milk proteins seem to form a structurally related protein family. The gene for the 64-kDa protein/RJP57 1 was expressed specifically in the nurse-bee gland, whereas that for the 56-kDa protein was expressed in both the nurse-bee and forager-bee glands. mRNAs for the 56-kDa and 64-kDa proteins were detected by in situ hybridization in a whole acinus of the nurse-bee gland, whereas mRNAs for the 56-kDa protein and alpha glucosidase were detected in that of the forager-bee gland. Therefore, the individual secretory cells of the acinus of the hypopharyngeal gland were shown to express these genes differently with the age-dependent role change of the worker bee. PMID- 9395330 TI - Human endothelin receptors ET(A) and ET(B) expressed in baculovirus-infected insect cells--direct application for signal transduction analysis. AB - We expressed human endothelin receptors, ET(A) and ET(B), in insect Sf9 cells infected by recombinant baculoviruses that contained the respective cDNAs. Ligand binding experiments showed that the two expressed receptors have the same affinities as observed for the receptors in mammalian cells, i.e. the ET(A) receptor showed an affinity order of ET-1 > or = ET-2 >> ET-3, and the ET(B) receptor remained nonselective for three isopeptide ligands. The ET(B) receptor was purified by affinity chromatography with K9-biotinyl-ET-1 without losing the ligand-binding activity from the membrane of infected Sf9 cells. Protein chemical analysis of the purified ET(B) receptor showed that it is glycosylated, and that the N-terminal 38-amino-acid peptide is susceptible to proteolytic digestion, resulting in a small 35-kDa receptor like that found in the human placenta. Surprisingly, the infected and unlysed cells showed a strong intracellular Ca2+ concentration increase ([Ca2+]i), which was generated by a unique signal transduction pathway consisting of the insect GTP-binding protein and human endothelin receptors expressed in the late phase of virus infection. Due mainly to an efficient expression (over 200,000 receptors/cell), to a low background owing to no endogenous homolog receptor in insect Sf9 cells, and to a sensitive fluorescent reagent Fura-2, this insect Sf9 cell system can detect the [Ca2+]i induced by picomolar levels of endothelin-receptor. We propose that this highly sensitive system be used to screen for potential antagonists/agonists of endothelin receptors. PMID- 9395331 TI - Effects of brefeldin A on phosphatidylcholine phospholipase D and inositolphospholipid metabolism in HL-60 cells. AB - The involvement of the small GTP-binding protein ADP-ribosylation factor (ARF) in guanosine 5'-[gamma-thio]triphosphate (GTP[S])-dependent activation of phospholipase D (PLD) in HL-60 cells has been well established in vitro. In this study, we tested the effect of brefeldin A, which prevents ARF activation by inhibiting guanine-nucleotide-exchange activity, on PLD stimulation by receptor agonists (formyl-Met-Leu-Phe and ATP) and by the phorbol ester phorbol 12 myristate 13-acetate (PMA) in differentiated HL-60 cells. However, brefeldin A did not affect the activation of PLD at a time (1 h) when it eliminated the activity of the trans-Golgi enzyme galactosyltransferase. It also did not inhibit PLD activity in Golgi-enriched membranes treated with GTP[S] with or without ARF in vitro. However, longer times of brefeldin A treatment (>6 h), progressively and completely inhibited the activation of PLD by formyl-Met-Leu-Phe and partly inhibited (approximately 50%) the activation by PMA. In contrast, long-term brefeldin A treatment did not inhibit the effect of GTP[S] on PLD in permeabilized HL-60 cells. Long-term brefeldin A treatment completely inhibited inositol phosphate production in response to formyl-Met-Leu-Phe and ATP, indicating that it affected inositolphospholipid-specific phospholipase C activity. These data indicate that the rapid inhibitory effect of brefeldin A on Golgi function is not associated with inhibition of receptor-mediated or PMA mediated PLD activation in HL-60 cells. However, longer-term effects, presumably arising from the disruption of the Golgi, lead to a total inhibition of agonist activation of PLD and inositolphospholipid-specific phospholipase C. In summary, these results do not support a role for brefeldin-A-sensitive ARF in agonist regulation of PLD in HL-60 cells. PMID- 9395332 TI - Characterization, gene cloning and expression of isocitrate lyase involved in the assimilation of one-carbon compounds in Hyphomicrobium methylovorum GM2. AB - Isocitrate lyase of an obligate methylotrophic bacterium, Hyphomicrobium methylovorum GM2, was purified to homogeneity and characterized. The enzyme is a homotetramer of identical 62-kDa subunits. After the enzyme had been incubated at temperatures up to 25 degrees C for 30 min, no loss of activity was observed. The enzyme was stable in the pH range of 7.5-9.0. Maximum activity was observed at pH 7.5 and around 45 degrees C. The Km value for Ds-isocitrate was 0.51 mM. The activity required Mg2+ and was inhibited by oxalate, succinate and glycolate. The gene encoding the isocitrate lyase and its flanking regions were isolated from H. methylovorum GM2. Nucleotide sequencing of recombinant plasmids revealed that the isocitrate lyase gene codes for a 540-amino-acid protein. The amino acid sequence of the enzyme is similar to those of the enzymes from Escherichia coli (40% identity) and cucumber (37% identity). The recombinant plasmid, which was constructed by ligation of the cloned gene and an expression vector, pKK223-3, was introduced into E. coli HB101. The transformed E. coli cells expressed isocitrate lyase, which was indistinguishable from the purified H. methylovorum GM2 isocitrate lyase on analysis by SDS/PAGE. PMID- 9395333 TI - Neuronal cell nuclear factor--a nuclear receptor possibly involved in the control of neurogenesis and neuronal differentiation. AB - We have cloned from a cDNA library of neuronal derivatives of retinoic-acid induced embryonic carcinoma cells a nuclear receptor that may be involved in the control of late neurogenesis and early neuronal differentiation. The receptor which is practically identical in sequence with germ cell nuclear factor, has been designated neuronal cell nuclear factor (NCNF). NCNF is exclusively expressed in the neuronal derivatives of PCC7-Mz1 cells, with the expression beginning within hours of exposure to retinoic acid. In the developing mouse brain, NCNF is expressed in the marginal zones of the neuroepithelium which are known to contain young postmitotic neurons. NCNF binds to the DR0 sequence thereby silencing transcription. Because NCNF does not recognize hormone response elements of other nuclear receptors tested and does not heterodimerize with these, it probably binds exclusively as a homodimer. NCNF may induce neuronal differentiation by repressing the activity of genes that permit cell fates other than the neuronal one. PMID- 9395334 TI - Characterization and evolution of a gene encoding a Trimeresurus flavoviridis serum protein that inhibits basic phospholipase A2 isozymes in the snake's venom. AB - The proteins that bind phospholipase A2 (PLA2) isozymes of Trimeresurus flavoviridis (habu snake, crotalinae) venom were fractionated from sera on four columns, each conjugated with one of four PLA2 isozymes. Five proteins, termed PLA2 inhibitors (PLI) I-V, were obtained as the binding components. The combinations of the binding components differed depending on the PLA2 isozymes. PLI-IV and PLI-V correspond to PLI-A and PLI-B, respectively, which were known to bind to a major [Asp49]PLA2, PLA2, and contained a segment similar to the carbohydrate-recognition domain of C-type lectins. PLI-I, which is a major component of inhibitory proteins against three basic PLA2 isozymes, PLA-B (a basic [Asp49]PLA2) and basic proteins I and II (both [Lys49]PLA2s), has been isolated, and its partial amino acid sequence has been determined. A cDNA encoding PLI-I was isolated from a T. flavoviridis liver cDNA library and sequenced. PLI-I cDNA encoded 200 amino acid residues, including a signal peptide of 19 amino acid residues. One sugar chain was predicted to occur at position 157. A gene coding for PLI-I was isolated. It is 9.6-kb long and consists of five exons and four introns. Comparison of the exon-intron structure of the PLI-I gene with those of genes encoding urokinase-type-plasminogen-activator receptor (uPAR), Ly-6, CD59 and neurotoxins showed that they have characteristic unit encoding approximately 90 amino acid residues, which is divided over two exons. This strongly suggests that the PLI-I gene belongs to the uPAR, Ly-6, CD59 and neurotoxin gene family. There are two types of structurally different inhibitors against PLA2 isozymes in T. flavoviridis serum with different evolutionary origins. PMID- 9395335 TI - Inhibitory guanine-nucleotide-binding-regulatory protein alpha subunits in medaka (Oryzias latipes) oocytes--cDNA cloning and decreased expression of proteins during oocyte maturation. AB - We have previously shown that pertussis-toxin-sensitive inhibitory guanine nucleotide-binding-regulatory proteins (G proteins) are involved in the signal transduction of steroidal maturation-inducing hormone (MIH) of rainbow trout (Oncorhynchus mykiss) oocytes, 17alpha,20beta-dihydroxy-4-pregnen-3-one (17alpha,20beta-DP) [Yoshikuni, M. & Nagahama, Y. (1994) Dev. Biol. 166, 615 622]. In this study, we obtained five different cDNA fragments of G protein alpha subunits from medaka (Oryzias latipes) intact ovarian follicles (three subtypes of G(i alpha), G(i alpha a), G(i alpha b) and G(i alpha c); two subtypes of G(s alpha), G(s alpha d), and G(s alpha e)). Using a newly developed extraction method for medaka oocyte RNA, we demonstrated that oocytes expressed both G(i alpha a) and G(i alpha c), but not G(i alpha b). Full-length cDNA clones for G(i alpha a) and G(i alpha c) were then isolated from a medaka ovarian follicle cDNA library. The predicted amino acid sequences of G(i alpha a) and G(i alpha c) exhibited significant similarity with G(i alpha1) and G(i alpha2) of other species, respectively. Both G(i alpha a) and G(i alpha c) possessed a specific Cys residue in the C-terminal region that was the site for ADP-ribosylation by pertussis toxin. G(o alpha), another G protein that is ADP-ribosylated by pertussis toxin, was not detected in oocytes, although it was expressed in brain tissue. Western blot analyses using a specific antibody against G(i alpha1) and G(i alpha2) subunit proteins revealed that in both medaka and rainbow trout G(i alpha) subunit protein (40 kDa) contents were abundant in plasma membranes of postvitellogenic immature oocytes, decreased in mature oocytes, and were absent in ovulated eggs. Furthermore, specific 17alpha,20beta-DP binding to plasma membranes was higher in postvitellogenic immature oocytes than in ovulated eggs. Taken together, these results suggest that G(i alpha a) and/or G(i alpha c) may be involved in the transduction of the signal from 17alpha,20beta-DP receptors during oocyte maturation of fish oocytes. PMID- 9395336 TI - Isolation and characterization of goldfish Y box protein, a germ-cell-specific RNA-binding protein. AB - Cyclin B is a regulatory subunit of maturation-promoting factor. In goldfish (Carassius auratus) oocytes, cyclin B is synthesized de novo after stimulation by 17alpha,20beta-dihydroxy-4-pregnen-3-one (maturation-inducing hormone). In this study, we examined goldfish oocyte proteins bound to cyclin B mRNA. Using oligo(dT)-cellulose affinity chromatography and northwestern blotting analysis, we identified a 54-kDa cyclin B mRNA-binding protein (p54). Southwestern blotting analysis showed the binding of p54 to the Y box DNA element (CTGATTGGCCAA), suggesting that p54 is a Y box protein in goldfish. We isolated two cDNA clones, GFYP1 and GFYP2, the latter of which encodes a germ-cell-specific Y box protein. An antibody against a GFYP2 protein recognized p54, suggesting that p54 is identical or highly similar to GFYP2 protein. This is also supported by the finding that a recombinant GFYP2 expressed in bacteria bound to both the Y box DNA element and the goldfish cyclin B mRNA synthesized in vitro. These results suggest that p54 is a germ-cell-specific Y box protein and is a potential masking protein of cyclin B mRNA in goldfish oocytes. PMID- 9395337 TI - Supramolecular assemblies from lysosomal matrix proteins and complex lipids. AB - Most lysosomal hydrolases are soluble enzymes. Lamp-II (lysosome-associated membrane protein-II) is a major constituent of the lysosomal membrane. We studied the aggregation of a series of lysosomal molecules. The aggregation-sensitive lysosomal marker enzymes were optimally aggregated at intralysosomal pH. A similar pH dependence was recorded for aggregation of Lamp-II. The pH-dependent loss of solubility of isolated Lamp-II required components of the lysosome extract. Conditions of mild acid pH promoting aggregation triggered the formation of complexes with lipids of lysosomal origin. We fractionated a membrane-free lysosome extract by gel-filtration chromatography and could reconstitute assemblies in vitro from separated fractions. We found some selectivity in the lysosomal proteins binding to complex lipids, phosphatidylcholine, sphingomyelin, and phosphatidylethanolamine being most effective. We propose that the formation at pH 5.0 of such supramolecular assemblies between lysosomal proteins and lipids occurs within the intralysosomal environment. Some possible consequences of such an intralysosomal matrix formation on organelle function are discussed. PMID- 9395338 TI - Analysis of the complex formed by Erythrina variegata chymotrypsin inhibitor with chymotrypsin and properties of the peptides prepared from the inhibitor by a limited proteolysis. AB - The stoichiometry of Erythrina variegata chymotrypsin inhibitor (ECI) and chymotrypsin interaction was previously estimated to be 1:2 by a titration of inhibitory activity. In the present study, gel-permeation chromatography and reverse-phase HPLC (RP-HPLC) were employed to analyze the complex formed by the inhibitor and enzyme. The results showed that ECI and chymotrypsin molecules undergo aggregation in the complex-forming buffer simultaneously with a binary complex consisting of one ECI and one chymotrypsin molecules in a soluble form. A mild lysylendopeptidase digestion of ECI produced two peptides in high yield, which were separated by RP-HPLC and characterized in terms of their structures and inhibitory activities. The N-terminal peptide, ECI-(1-107)-peptide, containing the primary reactive site retained a slight inhibitory activity, while the C-terminal peptide, ECI-(108-179)-peptide, exhibited no inhibitory activity. The inhibitory potency of the ECI-(1-107)-peptide was enhanced by the presence of the ECI-(108-179)-peptide in reconstituted mixture. Recovery of the native-like structure of the reconstituted complex was further indicated by fluorescence spectra, which showed strong conformational interaction between the two peptides; their dissociation constant Kd was calculated to be 209 nM. Taken together with the previous result obtained by chymotryptic digestion, it is suggested that the primary binding loop in ECI interacts with chymotrypsin not only by a standard mechanism but also by a non-substrate-like manner. Alternatively, ECI might have an additional binding segment in the N-terminal region which interacts with chymotrypsin by a non-substrate-like manner. Further, it is shown that the C terminal region may support the native conformation of the binding loop(s) in the N-terminal region as an intramolecular chaperone. PMID- 9395339 TI - Comparative study of chicken and human parathyroid hormone-(1-34)-peptides in solution with SDS. AB - Molecular conformations of chicken [cPTH-(1-34)] and human [hPTH-(1-34)] parathyroid hormone fragments in aqueous solutions with various concentrations of SDS were investigated by CD, fluorescence and NMR spectroscopy techniques. In the presence of SDS, chicken and human PTH-(1-34) adopt an a-helical structure making up 32-38% of all the peptide amino acids. The process of the a-helical formation of these two fragments is considerably different. The CD spectral change of hPTH (1-34) was characteristic of a monotonous increase in the negative peak at 222 nm with increasing SDS concentrations. However, for cPTH-(1-34) a beta-turn is formed first, followed by alpha-helix formation upon an increase in SDS concentrations. The change of the tryptophan fluorescence spectra of cPTH-(1-34) is well correlated with the changes in CD spectra, suggesting that the side chain of Trp23 is involved in the conformational change from random coil to alpha-helix via beta-turn. The three-dimensional structure of cPTH-(1-34) with SDS micelle was elucidated by 1H-NMR at pH 3.8 and 300 K, with the combined use of distance geometry and restrained molecular dynamics calculations. NMR results indicated that it contains two helices encompassing residues 7-12 and 24-30, respectively. The C-terminal helix in the residue range of 24-30 is amphiphilic, which is stabilized by the hydrophobic interactions among Trp23, Leu24 and Lys27. PMID- 9395340 TI - Biochemical and immunological characterization of recombinant allergen Lol p 1. AB - Pollen from perennial rye grass (Lolium perenne), a major cause of type-I allergy worldwide, contains a complex mixture of allergenic proteins among which Lol p 1 is one of the most important. We describe the expression, purification and characterization of a recombinant Lol p 1 overproduced in Escherichia coli. The recombinant allergen, expressed in high yields and purified in milligram amounts, bound to specific IgE antibodies from human sera, induced histamine release from sensitized human basophils, and elicited rabbit antisera that recognize specifically recombinant Lol p 1 and natural Lol p 1 of pollen extract. Recombinant Lol p 1 was used to develop ImmunoCAP assays for analysis of 150 sera that were Radioallergosorbent test positive to L. perenne pollen. In 130 of them (87%) the assay detected a significant level of IgE antibodies to Lol p 1, reaching on average 37% of the level obtained with a test for IgE to the whole grass pollen extract. To map epitopes on Lol p 1, we produced three deletion mutants [des-(116-240)-Lol p 1, des-(1-88)-Lol p 1 and des-(133-189)-Lol p 1], which were efficiently expressed in bacteria. These all showed a strong reactivity with the specific rabbit IgG antibodies, but lacked most or all the allergenic properties of recombinant Lol p 1. A study of the antigenic structure of Lol p 1 was performed using the three deletion mutants and a set of 17-18 residue overlapping synthetic peptides covering the whole allergen sequence. The results indicate that human IgE and rabbit IgG antibodies bind to distinct regions of Lol p 1, and that at least some important IgE epitopes are mainly conformational. The findings suggest that recombinant allergens constitute useful reagents for further development of serological diagnosis of allergy, and that it should be possible to produce immunogenic fragments of allergenic proteins without allergenic properties. PMID- 9395341 TI - Structural properties of chimeric peptides containing a T-cell epitope linked to a fusion peptide and their importance for in vivo induction of cytotoxic T-cell responses. AB - We have previously shown that when administered to mice without adjuvant, a chimeric peptide consisting of the fusion peptide F from measles virus protein linked at the C-terminus of a cytotoxic T-cell epitope from the M2 protein of respiratory syncytial virus efficiently primes for an major histocompatibility complex (MHC) class-I restricted cytotoxic T lymphocyte (CTL) response. In this report, we demonstrated by microspectrofluorometry that the fusion-peptide moiety bound to the plasma membrane of living cells. When the fusion peptide was linked to the C-terminus of the CTL epitope, the chimeric peptide (M2-F) adopted a marked beta-sheet conformation. In contrast, when the fusion peptide was linked to the N-terminus of the T-cell epitope (F-M2), the chimeric peptide adopted an alpha-helical conformation in the presence of trifluoroethanol. The immunogenicity of the two chimeric peptides for class-I restricted CTL was also significantly different, the one adopting the alpha-helical conformation being more immunogenic. Probably due to its obvious conversion to an alpha-helical conformation, the F-M2 peptide could have a higher propensity to insert into membranes, as shown by microspectrofluorometry, with a resultant better immunogenicity than the M2-F peptide. PMID- 9395342 TI - The cell wall polymer of the extremely halophilic archaeon Natronococcus occultus. AB - The cell wall polymer of Natronococcus occultus (DSM 3396) consists of L glutamate, N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, D-galacturonic acid, D-glucuronic acid and D-glucose in a molar ratio of 5:7:1:8:0.5:0.3. Partial acid hydrolysis of the cell wall polymer produced soluble fragments that could be separated by HPLC. A gamma-glutamyl dipeptide was isolated. In the intact cell wall polymer, the glutamate residues form a poly-(gamma-glutamine) chain with a length of about 60 monomers, which corresponds to a relative molecular mass of approximately 7700 Da. Two other soluble dimeric fragments, composed of glutamate and either glucosamine or galactosamine in a molar ratio of 1:1, were purified from the hydrolysate, suggesting the presence of two different oligosaccharides linked to the poly-(gamma-glutamine) chain of the intact polymer. The analysis of additional fragments, which were composed of an amino sugar and galacturonic acid or glucose indicated that one oligosaccharide consisted of a glucosamine pentamer in an alpha-1,3 linkage at the reducing end and an oligomer with at least five beta-1,4-linked galacturonic acid residues at the non-reducing end. The second oligosaccharide was comprised of a galactosamine dimer in a beta-1,3 linkage at the reducing end and a maltose unit at the non-reducing end. Both oligosaccharides were linked to the alpha-amide group of the glutamine residues of the poly-(gamma-glutamine) chain. The whole cell wall polymer, which represents a novel type of natural glycoconjugate, has a relative molecular mass of 54 kDa. PMID- 9395343 TI - Overlay versus underlay tympanoplasty. Part I: historical review of the literature. AB - The history of otology is the history of the successful treatment of infections of the middle ear and the eardrum. Otologists have sought to restore hearing lost to infections of the eardrum since the 1600s. The development of instruments, techniques, and materials to treat infection is fascinating because of the serendipitous nature of the discoveries and the insight of the discoverers. This historical review describes the history of the treatment of infections of the ear and the development of modern techniques of ear surgery. Two contemporary methods of tympanic membrane repair are then described. PMID- 9395344 TI - Overlay versus underlay tympanoplasty. Part II: the study. AB - This report compares two contemporary techniques of tympanic membrane repair. The prospective comparison study included 712 cases over 9 years. Whether the tympanic membrane was repaired by an underlay or an overlay technique, results were reliable. By making a postauricular incision, greater visibility of the operative site can be obtained. Larger perforations can be closed more reliably when greater exposure is obtained. The placement of the graft above or below the annulus is not the issue. Careful technique and precise work are the keys to successful tympanoplasty. Thus otologic surgeons should cultivate effective techniques, attempting to continuously improve their results to achieve perfection. PMID- 9395345 TI - Factors influencing the relapse of patients with inflammatory bowel disease. AB - The management of relapse of inflammatory bowel disease (IBD) remains a clinical challenge and a relatively neglected area of current research. Many factors contribute to relapse, and the proper identification of the cause of each case may influence optimal management. Often, relapse is related to the failure of maintenance therapy. Mesalamine sensitivity is difficult to recognize and should be considered when increased doses are associated with the worsening of symptoms. An increase in eosinophil activity could explain seasonal relapses of IBD as a result of exposure to allergens, but other eosinophil activation pathways also will influence the course of IBD. Infection, either enteric or systemic, may trigger a relapse by activating the gastrointestinal mucosal immune system. Nonsteroidal anti-inflammatory drug use is a well-recognized cause of exacerbation of disease. Smoking status is emerging as a complex factor in IBD, and a change in smoking status may influence the course of IBD. PMID- 9395346 TI - Pathogenesis and immune mechanisms of chronic inflammatory bowel diseases. AB - The inflammatory bowel diseases (IBDs) are characterized by intestinal inflammation of unknown etiology. Two distinct disorders, Crohn's disease and ulcerative colitis, have been identified. Three theories of IBD etiology are currently under consideration: 1) reaction to a persistent intestinal infection, 2) existence of a defective mucosal barrier to luminal antigens, and 3) a dysregulated host immune response to ubiquitous antigens. In each of these theories, either pathogenic or resident luminal bacteria constantly stimulate the mucosal and systemic immune systems to perpetuate the inflammatory cascade. Chronicity of inflammation results from an interaction of the persistent stimulus of microbial antigens with genetically determined host susceptibility factors that determine the individual's immune response or mucosal barrier function. The pathogenesis of IBD involves a series of steps, beginning with the breach of the intestinal mucosal barrier by infectious agents or toxins. The defective barrier exposes lamina propria immune cells to the continual presence of resident luminal bacteria, bacterial products, or dietary antigens, which perpetuates the inflammatory cascade. Many immunoregulatory abnormalities are noted in IBD, including the ratio of proinflammatory to immunosuppressive cytokines, selective activation of T(H) lymphocyte subsets, and abnormalities in epithelial antigen presentation. When activated during the initial inflammatory process, macrophages and T lymphocytes secrete a host of cytokines, which recruit other inflammatory cell types, thereby continuing the process. Tissue injury is the net result of the soluble products of the activated inflammatory cells. Knowledge of the pathogenesis in IBD suggests that the ultimate goals of therapy should be to block the proinflammatory mediators toward the proximal, rather than the distal, end of the cascade, to decrease the constant antigenic drive of luminal bacteria, and to correct the dysregulated immune response. PMID- 9395347 TI - Optimizing therapy for inflammatory bowel disease. AB - This review focuses on current developments in the major categories of therapy used in the management of inflammatory bowel disease (IBD). Conventional corticosteroids, although a mainstay of the acute treatment of IBD for many years, have many drawbacks, including a variety of side effects--particularly with chronic use. Budesonide appears to be relatively safe and at least moderately effective in inducing remission in active distal ulcerative colitis (UC) and Crohn's disease. Aminosalicylates, both oral and topical, have proven useful in managing mild-to-moderate active UC and mild active Crohn's disease, as well as in maintaining remission. Data from recent trials suggest that higher doses of mesalamine are generally more efficacious than lower doses. In addition, a combination of oral and rectal formulations may succeed when one route, alone, is not successful. The immunomodulatory agents azathioprine, 6-mercaptopurine, and methotrexate have been shown to be effective in the treatment of IBD and are now widely accepted as valuable parts of the therapeutic armamentarium. Cyclosporine, although effective, is associated with many toxicities, and patients must be monitored closely in centers experienced with this agent. Clinical trials of IL-10, IL-11, and anti-TNFalpha have also shown promise. Antibiotics have been used empirically for many years in the treatment of IBD. Larger clinical trials are warranted to explore the potential efficacy of antibiotic therapy. This has been accomplished with metronidazole in Crohn's disease, and other antibiotic trials are underway at this time. The investigational agents acemannan, heparin, and transdermal nicotine have also shown variable degrees of promise as possible therapies for IBD. Despite the variety of agents available for the treatment of IBD, none is ideal or universally accepted. Ongoing research into the well-established therapeutic agents, as well as novel drugs with more precise targets, may contribute to the design of a more nearly optimal regimen for IBD in the not-too-distant future. PMID- 9395348 TI - Quality of life issues in patients with inflammatory bowel disease. AB - The assessment of health-related quality of life (HRQOL) plays an increasingly important role in the evaluation of therapeutic interventions in various chronic diseases, including inflammatory bowel disease (IBD). There are three main types of HRQOL instruments, each of which have specific strengths and limitations. Global assessments provide a general impression of the patient's HRQOL, but they do not identify specific domains of dysfunction. Generic instruments can be used to show similarities or differences among groups or populations, but they may not be sensitive to changes over time or after treatment in groups of patients with specific diseases. Disease-specific instruments, which are derived from and validated in particular disease groups, are the most sensitive indicators of change in HRQOL over time or with treatment. There are several potential sources of bias in HRQOL measurement, and certain instruments are more or less sensitive and reliable than others in detecting impaired function in patients with IBD. Despite differences between studies regarding specific HRQOL findings, as a group, these assessments confirm that HRQOL can be impaired significantly in patients with IBD. Reliable validated HRQOL instruments should be applied with the same methodological rigor as other assessments in clinical trials of different therapeutic strategies for patients with IBD. PMID- 9395350 TI - Pediatric hospitalizations for croup (laryngotracheobronchitis): biennial increases associated with human parainfluenza virus 1 epidemics. AB - Croup is a common manifestation of respiratory tract infection in children, and human parainfluenza virus 1 (HPIV-1) is the agent most commonly associated with croup. In the United States, HPIV-1 produces a distinctive pattern of biennial epidemics of respiratory illness during the autumn months of odd-numbered years. National Hospital Discharge Survey data for croup hospitalizations among patients <15 years old between 1979 and 1993 were examined along with laboratory-based surveillance data on HPIV-1 activity in the United States. The mean annual number of croup hospitalizations was 41,000 (range, 27,000-62,000/year). Ninety-one percent of hospitalizations occurred among children <5 years of age. Minor peaks in croup hospitalizations occurred each year in February, and major peaks occurred in October of odd-numbered years, coincident with peak HPIV-1 activity. Each biennial epidemic of HPIV-1 was associated with 18,000 excess croup hospitalizations nationwide. PMID- 9395349 TI - Effects of the neuraminidase inhibitor zanamavir on otologic manifestations of experimental human influenza. AB - Middle ear pressure (MEP) abnormalities are frequently observed during influenza virus infection and may serve as surrogate markers for the risk of otitis media. MEP abnormalities were evaluated in adult volunteers who were inoculated with influenza A/Texas/36/91(H1N1) or B/Yamagata/88 virus and given the antiviral zanamivir (GG167) intranasally as prophylaxis or early treatment in randomized, double-blind, placebo-controlled trials. In the influenza A prophylaxis studies, 15% of 61 zanamivir recipients versus 61% of 33 placebo recipients showed significant MEP abnormalities (P < .01). In the influenza A early treatment trial, 32% of 31 infected zanamivir recipients versus 73% of 26 infected placebo recipients developed MEP abnormalities (P < .01). In the influenza B prophylaxis trial, 16% of 25 zanamivir versus 44% of 9 placebo recipients showed abnormalities (P = .09). These findings indicate that the neuraminidase inhibitor zanamivir, which is effective in reducing experimental influenza illness, provides protection against the development of MEP abnormalities. PMID- 9395351 TI - Evaluation of two live, cold-passaged, temperature-sensitive respiratory syncytial virus vaccines in chimpanzees and in human adults, infants, and children. AB - Two live-attenuated, cold-passaged (cp), temperature-sensitive (ts) candidate vaccines, designated cpts530/1009 and cpts248/955, were attenuated, genetically stable, and immunogenic in chimpanzees and were highly attenuated for human adults. In respiratory syncytial virus (RSV)-seropositive children, cpts530/1009 was more restricted in replication than cpts248/955. In seronegative children, 10(4) pfu of cpts248/955 was insufficiently attenuated, and a high titer of vaccine virus was shed (mean peak titer, 10(4.4) pfu/mL), whereas 10(4) pfu of cpts530/1009 was relatively attenuated and restricted in replication (mean peak titer, 10(2.0) pfu/mL). At a dose of 10(5) pfu, cpts530/1009 was immunogenic in seronegative children (geometric mean titer of RSV neutralizing antibodies, 1:724). Transmission of either vaccine to seronegative placebo recipients occurred at a frequency of 20%-25%. Of importance, vaccine viruses recovered from chimpanzees and humans were ts. In contrast to previous studies, this study indicates that live attenuated RSV vaccines that are immunogenic and phenotypically stable can be developed. Additional studies are being conducted to identify a live RSV vaccine that is slightly more attenuated and less transmissible than cpts530/1009. PMID- 9395352 TI - Adverse effects of maternal enterovirus infection on the fetus and placenta. AB - Gestational outcome in a murine model of congenital enterovirus infection was evaluated. Pregnant mice were inoculated intravenously with Theiler's murine encephalomyelitis virus (TMEV), a murine enterovirus, or with BHK 21 cell lysate (control) at 6-7 days of gestation (early) and sacrificed 6 or 12 days later, and their placentas and fetuses were studied. High rates of gross and histologic abnormalities (50%-87%) were seen in placentas and fetuses from dams infected with TMEV and sacrificed 6 days later. TMEV-infected dams sacrificed 12 days after inoculation had lower rates of placental-fetal abnormalities (25%-57%) but an additional 42% rate of complete pregnancy loss. Pregnancy loss (9%) and placental-fetal abnormalities (4%-7%) were uncommon in control animals. Rates of fetal abnormalities and placental infection in infected dams exceeded fetal viral infection, suggesting that TMEV infection adversely affects pregnancy either directly by fetal damage or indirectly by placental compromise. PMID- 9395354 TI - Anti-interleukin-10 antibodies in patients with chronic active Epstein-Barr virus infection. AB - The Epstein-Barr virus (EBV) genome encodes a protein in its BamHI C restriction fragment rightward open-reading frame-1 (designated BCRF1 or viral interleukin-10 [vIL-10]) that shares protein homology and biologic properties with human IL-10. Several EBV disorders are characterized by prolonged active EBV infection. Because continued EBV replication could allow for increased vIL-10, ELISA and immunoprecipitation were used to determine whether vIL-10 expression during chronic active EBV infection resulted in vIL-10 and IL-10 antibodies. IL-10 antibodies were assayed in patients diagnosed with chronic and acute infectious mononucleosis (CIM, AIM), nasopharyngeal carcinoma (NPC), and EBV-associated lymphoproliferative disease (LPD), as well as from healthy organ transplant patients and EBV-negative or EBV-positive persons. Whether anti-IL-10 antibodies could inhibit IL-10 biologic activity was determined. vIL-10 antibodies were found in CIM, NPC, and LPD patients and antibodies reactive to IL-10 were found in CIM patients. One CIM patient had IL-10 antibodies that neutralized IL-10 bioactivity in vitro. PMID- 9395353 TI - Recombinant bacille Calmette-Guerin expressing the measles virus nucleoprotein protects infant rhesus macaques from measles virus pneumonia. AB - Measles virus infection continues to be a major cause of infant mortality. There is a need for a measles vaccine that can be administered at birth in the presence of maternal neutralizing antibody. Infant rhesus monkeys were immunized with recombinant bacille Calmette-Guerin expressing the full-length measles virus nucleoprotein (BCG-N) and subsequently challenged with measles virus. Nucleoprotein-specific lymphocyte proliferative responses were detected in the absence of anti-N antibody after vaccination. Vaccination with BCG-N did not prevent systemic measles virus infection; however, there was a significant reduction of lung inflammation after challenge. Virus titers in lymph nodes were significantly lower, and the duration of nasopharyngeal viral shedding was shorter in some vaccinated monkeys after challenge. These results suggest that measles virus-specific T cells were primed by BCG-N vaccination and that they prevented virus-induced lung pathology. PMID- 9395355 TI - Posttransplant lymphoproliferative disease in primary Epstein-Barr virus infection after liver transplantation: the role of cytomegalovirus disease. AB - Epstein-Barr virus (EBV) plays a major role in the pathogenesis of posttransplant lymphoproliferative disease (PTLD). Patients who undergo primary EBV infection after transplantation are at greater risk of developing PTLD. In this retrospective study, the incidence of EBV infection and associated PTLD in 40 consecutive adult recipients who were seronegative for EBV at the time of liver transplantation were investigated, and risk factors for PTLD were analyzed. Of 37 patients with available timely posttransplant serum samples, 35 (95%) developed primary EBV infection. Of the 40 patients, 13 (33%) developed PTLD a median of 126 days (range, 48-776) after liver transplantation. The factor significantly associated with the development of PTLD was cytomegalovirus disease (relative risk, 7.3; 95% confidence interval, 2.36-22.6; P = .0006). Cytomegalovirus disease is a predictor for the development of PTLD in primary EBV infection after liver transplantation, and it may be a target for prophylactic intervention. PMID- 9395356 TI - Increased prevalence of infectious diseases and other adverse outcomes in human T lymphotropic virus types I- and II-infected blood donors. Retrovirus Epidemiology Donor Study (REDS) Study Group. AB - Disease associations of human T lymphotropic virus types I and II (HTLV-I and II) infection were studied in 154 HTLV-I-infected, 387 HTLV-II-infected, and 799 uninfected blood donors. Adjusted odds ratios (ORs) and 99% confidence intervals (CIs) were derived from logistic regression models controlling for demographics and relevant confounders. All subjects were human immunodeficiency virus type 1 seronegative. HTLV-II was significantly associated with a history of pneumonia (OR, 2.6; 99% CI, 1.2-5.3), minor fungal infection (OR, 2.9; 99% CI, 1.2-7.1), and bladder or kidney infection (OR, 1.6; 99% CI, 1.0-2.5) within the past 5 years and with a lifetime history of tuberculosis (OR, 3.9; 99% CI, 1.3-11.6) and arthritis (OR, 1.8; 99% CI, 1.2-2.9). Lymphadenopathy (> or =1 cm) was associated with both HTLV-I (OR, 6.6; 99% CI, 2.2-19.2) and HTLV-II (OR, 2.8; 99% CI, 1.1 7.1) infection, although no case of adult T cell leukemia/lymphoma was diagnosed. Urinary urgency and gait disturbance were associated with both viruses. This new finding of increased prevalence of a variety of infections in HTLV-II-positive donors suggests immunologic impairment. PMID- 9395358 TI - Interrelationships among quantity of human cytomegalovirus (HCMV) DNA in blood, donor-recipient serostatus, and administration of methylprednisolone as risk factors for HCMV disease following liver transplantation. AB - Longitudinal analysis of 162 liver transplant recipients identified 51 patients who were viremic. Virus load was determined in 47 of these patients using quantitative-competitive polymerase chain reaction. Peak virus load was significantly higher in 20 symptomatic patients than 27 asymptomatic patients (P < .0001). Elevated virus load, donor seropositivity, and total methylprednisolone dosage were risk factors for human cytomegalovirus (HCMV) disease (odds ratio [OR], 2.22/0.25 log10 increase in virus load, P = .001; OR, 4.11, P = .05; OR, 1.30/1-g increment in methylprednisolone, P = .01). Methylprednisolone and virus load were independent risk factors in a multivariate analysis (OR, 2.70/1-g increase, P = .003; OR, 1.61/0.25 log10 increase, P = .03, respectively). Virus loads of 10(4.75)-10(5.25) genomes/mL of blood were associated with an increased disease probability; the latter was shifted to lower virus loads with increasing quantities of methylprednisolone. These data illustrate the central role of virus load in HCMV pathogenesis. PMID- 9395357 TI - Distinguishing baboon cytomegalovirus from human cytomegalovirus: importance for xenotransplantation. AB - The severe shortage of human organs for transplantation is the driving force behind xenotransplant research. Nonhuman primates, particularly baboons, are potential sources of organs and tissues. Human cytomegalovirus (HCMV) is the most common donor-associated infection after allotransplantation. Baboon cytomegalovirus (BCMV) is endemic in baboon populations and therefore is a potential cause of donor-associated disease after xenotransplantation. Accordingly, the ability for BCMV to grow in human cells was determined and a sensitive method to distinguish BCMV from HCMV was developed. Human fibroblasts were permissive for BCMV, isolates exhibited cytopathology characteristic of HCMV, and herpesvirus-like virions were observed by electron microscopy. BCMV and HCMV could be distinguished by restriction fragment length polymorphism patterns and by polymerase chain reaction with primers targeting the BCMV major immediate early gene promoter. These methods can be used to evaluate BCMV pathogenicity in laboratory and clinical xenotransplant trials. PMID- 9395359 TI - Hepatitis G virus infection in American patients with cryptogenic cirrhosis: no evidence for liver replication. AB - It is unclear whether hepatitis G virus (HGV) can lead to chronic liver disease and cirrhosis. Eighty-nine patients with end-stage liver disease undergoing liver transplantation were studied: 50 were diagnosed as having cryptogenic cirrhosis while 39 had nonviral chronic liver disease. Five (10%) in the former and 1 (2.6%) in the latter group (not significantly different) were positive for HGV RNA in serum. All 6 HGV-infected patients were negative for the presence of the HGV RNA minus strand in the liver when tested with a strand-specific Tth-based reverse transcription-polymerase chain reaction, and 5 were positive for the presence of the plus strand, albeit at low levels. This implies that the liver is not the primary replication site for HGV, at least in a significant proportion of patients. Absence of liver replication explains the reported lack of association between HGV infection and liver pathology encountered in many clinical settings. PMID- 9395360 TI - Varicella-zoster virus infection in children with underlying human immunodeficiency virus infection. AB - This article describes a prospective longitudinal study of varicella-zoster virus (VZV) infections in human immunodeficiency virus (HIV)-infected children, designed to determine their natural history of VZV infection and possible effects of VZV on the progression of HIV infection. Varicella was usually not a serious acute problem, and it did not seem to precede clinical deterioration. The rate of zoster was high: 70% in children with low levels of CD4+ lymphocytes at the time of development of varicella. It is predicted that immunization with live attenuated varicella vaccine is unlikely to be deleterious to HIV-infected children. Moreover, if they are immunized when they still have relatively normal levels of CD4+ lymphocytes, they may have a lower rate of reactivation of VZV than if they were allowed to develop natural varicella when their CD4+ cell counts have fallen to low levels as a result of progressive HIV infection. PMID- 9395361 TI - DNA vaccination as anti-human immunodeficiency virus immunotherapy in infected chimpanzees. AB - The role of the immune response in controlling human immunodeficiency virus type 1 (HIV-1) replication is controversial. Immunotherapeutic strategies that have the ability to broaden immune responses might play a role in slowing disease progression. DNA immunization was studied as immunotherapy in infected chimpanzees. Two HIV-1-infected chimpanzees were vaccinated with DNA plasmid vaccines, one with plasmid pCMN160, which expresses the envelope glycoprotein of HIV-1MN and rev, and the other with a control plasmid. The chimpanzee immunized with pCMN160 demonstrated enhanced humoral responses. Virus load was monitored. Virus load in the chimpanzee receiving pCMN160 decreased at week 20 and has remained at background levels. The control chimpanzee was subsequently vaccinated with pCMN160. After immunization, the antibody responses increased and, as in the first animal, the virus load decreased. These results indicate the potential of the immune response to have a direct impact on HIV-1 replication in chimpanzees. PMID- 9395362 TI - Human immunodeficiency virus infection impairs hemopoiesis in long-term bone marrow cultures: nonreversal by nucleoside analogues. AB - Hematologic abnormalities are often seen in patients infected with human immunodeficiency virus (HIV). The effect of HIV infection of bone marrow stroma on support of uninfected CD34 progenitor cells in long-term bone marrow culture (LTBMC) was investigated. Results show that HIV-infected bone marrow stroma was unable to adequately support CD34 progenitor cells in vitro. Zidovudine or didanosine was added to cultures in an attempt to reverse the suppressive effects exerted by HIV and to determine whether such suppression was mediated by transfer of HIV infection to progenitor cells. Didanosine failed to reduce the suppressive effects of HIV, whereas zidovudine compounded the observed suppression. HIV infection of bone marrow stroma, while reducing the production of nonadherent cells, did not increase apoptosis and cell death in such cells. In contrast, zidovudine enhanced apoptosis and cell death in nonadherent cells produced by both HIV-infected and control LTBMC. PMID- 9395363 TI - The safety and efficacy of adefovir dipivoxil, a novel anti-human immunodeficiency virus (HIV) therapy, in HIV-infected adults: a randomized, double-blind, placebo-controlled trial. AB - Adefovir dipivoxil is a novel nucleotide analogue with several promising in vitro anti-human immunodeficiency virus (HIV) characteristics. To evaluate the safety and efficacy of adefovir dipivoxil monotherapy, a randomized, double-blind, placebo-controlled study was initiated involving 72 subjects with moderately advanced HIV disease. Subjects were randomly assigned in a 2:1 ratio to receive adefovir dipivoxil or placebo as a once-daily oral dose for 6 weeks, followed by 6 weeks of open-label adefovir dipivoxil. Two dose levels were studied (125 mg and 250 mg). Adefovir dipivoxil was determined to be safe and well-tolerated when administered for 12 weeks. At week 6, changes in absolute CD4 T cell levels and HIV-1 RNA levels were significantly greater with adefovir dipivoxil than with placebo. These effects were sustained through 12 weeks of treatment. As determined by standard RNA sequencing techniques, only 1 of the 24 subjects who received adefovir dipivoxil (125 mg/day) developed any genotypic change from baseline. PMID- 9395364 TI - Severe disease in children with trachoma is associated with persistent Chlamydia trachomatis infection. AB - The immediate study objective was to determine if variable disease severity in children with trachoma could be attributable in part to host variation in the ability to clear Chlamydia trachomatis infection. Identification of sibling cohorts with these variant phenotypes would be useful for immunogenetic studies. A weekly survey for 3 months in a trachoma-hyperendemic village using detection of chlamydial DNA and grading of disease severity indicated that 62% (33/53) of children had at least one infection episode. Of those, 64% (21/33) who were persistently infected had both significantly higher mean chlamydial DNA loads and more severe trachoma than did sporadically infected children. Of importance, duration of infection differed between siblings in 60% (6/10) of families. The results suggest that chlamydial load and duration of infection determine the chronic nature of severe disease in trachoma and that host variable efficiency for chlamydial clearance between siblings is in part determined by host variation. PMID- 9395365 TI - Delta-toxin from Staphylococcus aureus as a costimulator of human neutrophil oxidative burst. AB - Delta-toxin from Staphylococcus aureus is responsible for various pathophysiologic effects. By studying different cell types in binding of delta toxin in low, noncytotoxic concentrations, a specific binding of fluorescein labeled delta-toxin to neutrophils and monocytes was found. Studying direct effects of delta-toxin on neutrophils, a dose-dependent up-regulation of complement receptor 3 expression was found. Oxygen radical production, as determined by Luminol-enhanced chemiluminescence, was not directly induced by delta-toxin, and this toxin was also unable to prime neutrophils for an enhanced response to FMLP or complement-opsonized zymosan. However, the priming response induced by lipopolysaccharide or tumor necrosis factor-alpha (TNF-alpha) was significantly further enhanced in the presence of delta-toxin. Furthermore, as a direct effect on human monocytes, delta-toxin induced TNF-alpha production. These data provide evidence that delta-toxin has direct and indirect effects on the activity of neutrophils and monocytes with regard to its proinflammatory capacity. PMID- 9395366 TI - Predicting bacteremia in patients with sepsis syndrome. Academic Medical Center Consortium Sepsis Project Working Group. AB - The goal of this study was to develop and validate clinical prediction rules for bacteremia and subtypes of bacteremia in patients with sepsis syndrome. Thus, a prospective cohort study, including a stratified random sample of 1342 episodes of sepsis syndrome, was done in eight academic tertiary care hospitals. The derivation set included 881 episodes, and the validation set included 461. Main outcome measures were bacteremia caused by any organism, gram-negative rods, gram positive cocci, and fungal bloodstream infection. The spread in probability between low- and high-risk groups in the derivation sets was from 14.5% to 60.6% for bacteremia of any type, from 9.8% to 32.8% for gram-positive bacteremia, from 5.3% to 41.9% for gram-negative bacteremia, and from 0.6% to 26.1% for fungemia. Because the model for gram-positive bacteremia performed poorly, a model predicting Staphylococcus aureus bacteremia was developed; it performed better, with a low- to high-risk spread of from 2.6% to 21.0%. The prediction models allow stratification of patients according to risk of bloodstream infections; their clinical utility remains to be demonstrated. PMID- 9395367 TI - Inhibition of virulent Mycobacterium tuberculosis by Bcg(r) and Bcg(s) macrophages correlates with nitric oxide production. AB - The Nramp1 gene controls macrophage resistance or susceptibility to several intracellular microorganisms; however, there is conflicting evidence regarding its role during infection with virulent Mycobacterium tuberculosis. Nitric oxide (NO) is a potent antimycobacterial agent produced by macrophages, which is also regulated by Nramp1. The in vitro ability of B10R (resistant) and B10S (susceptible) murine macrophages to inhibit M. tuberculosis H37Rv and to produce NO in response to infection and interferon-gamma (IFN-gamma) was compared. Infected B10R macrophages inhibited [3H]uracil incorporation by M. tuberculosis and produced higher amounts of NO than did B10S macrophages. IFN-gamma increased the inhibitory activity of both cells. Inhibition of M. tuberculosis by IFN-gamma activated B10R macrophages was reversed by N(G)-monomethyl-L-arginine (N(G)MMA). L-arginine restored NO production and increased the antimycobacterial activity by IFN-gamma-stimulated N(G)MMA-treated macrophages. The Bcg/Nramp1 gene may regulate macrophage resistance or susceptibility to virulent M. tuberculosis by a differential capability of these cells to produce NO. PMID- 9395368 TI - Infection of swine with Mycobacterium bovis as a model of human tuberculosis. AB - Swine were infected with Mycobacterium bovis to develop a model for pulmonary and disseminated tuberculosis in humans. Pigs were inoculated with various doses of M. bovis by intravenous (i.v.), intratracheal (int), or tonsillar routes. Animals were euthanized between 17 and 60 days after inoculation, and tissues were collected for culture and histopathologic examination. Lesions of disseminated tuberculosis were found in pigs given 10(4) or 10(8) cfu of M. bovis i.v. or int; localized pulmonary disease was found in pigs given 10(2) or 10(3) cfu of M. bovis int. Lesions ranged from well-organized tubercles with coagulative necrosis, epithelioid macrophages, and fibrosis to large expansive tubercles with liquefactive necrosis and extracellular growth of M. bovis. Tuberculous meningitis was observed in animals given M. bovis i.v. Swine infected with M. bovis are a useful animal model for elucidating the mechanisms of pathogenesis and host defense to tuberculosis in humans. PMID- 9395370 TI - Aspergillus fumigatus conidia induce a Th1-type cytokine response. AB - The response of human peripheral blood mononuclear cells (MNC) to Aspergillus fumigatus in vitro was evaluated. In studies of the proliferative response of MNC from 18 healthy donors to heat-killed A. fumigatus conidia, 15 displayed a significant response, with a stimulation index (SI) between 4 and 193. In contrast, all donors displayed a positive response to Candida albicans blastoconidia (SI ranged from 10 to 224). Despite the variability in reactivity to A. fumigatus conidia, the response of a particular individual was stable when retested over periods of 1-2 weeks. Supernatant from cocultures of A. fumigatus conidia with MNC contained increased levels of interferon-gamma, granulocyte macrophage colony-stimulating factor, tumor necrosis factor-alpha, and interleukin (IL)-2, compared with unstimulated cells, but not IL-10 or IL-4. In addition, A. fumigatus induced lymphocyte surface expression of adhesion/activation-associated molecules. These results suggest that lymphocytes may contribute to host defense against Aspergillus by generating a Th1-type response. PMID- 9395369 TI - Mechanisms and target sites of damage in killing of Candida albicans hyphae by human polymorphonuclear neutrophils. AB - Target sites of fungal cell damage were studied to define mechanisms of neutrophil-mediated killing of Candida albicans hyphae. Neutrophils induced hyphal cell wall damage, as evidenced by release of cell wall glycoproteins and confocal microscopic changes. Damage occurred in the presence of neutrophil granule extracts and did not require oxidants. However, oxidation of hyphal surface glycoproteins correlated strongly with parallel increments in fungicidal activity, suggesting that oxidants did contribute to maximal cell wall damage. Neutrophil oxidants also induced hyphal DNA fragmentation, primarily single strand breakage, as shown by increased electrophoretic migration after nuclease S1 DNA digestion at single-strand break sites. The onset of damage to hyphal cell walls and DNA preceded detectable neutrophil-mediated fungicidal effects. Likewise, hyphal amino acid and nucleotide turnover as well as ATP initially rose, then declined as lethal effects became detectable. Thus, preceding detectable fungal cell death, neutrophil oxidative and oxygen-independent mechanisms damaged defined targets. PMID- 9395371 TI - Pathologic and clinical findings in patients with cyclosporiasis and a description of intracellular parasite life-cycle stages. AB - Cyclospora cayetanensis has been observed in the feces of persons with prolonged diarrhea. A description of the symptoms and histopathologic findings for patients with cyclosporiasis is presented. The intracellular life-cycle stages of these parasites in the enterocytes of patients will also be described. Seventeen Peruvian patients positive for Cyclospora organisms were surveyed and underwent endoscopy, and their symptoms were recorded. Patients presented with gastrointestinal symptoms, including diarrhea, flatulence, weight loss, abdominal discomfort, and nausea. Jejunal biopsies showed an altered mucosal architecture with shortening and widening of the intestinal villi due to diffuse edema and infiltration by a mixed inflammatory cell infiltrate. There was reactive hyperemia with vascular dilatation and congestion of villous capillaries. Parasitophorous vacuoles contained sexual and asexual forms. Type I and II meronts, with 8-12 and 4 fully differentiated merozoites, respectively, were found at the luminal end of epithelial cells. These findings demonstrate the complete developmental cycle associated with host changes due to Cyclospora organisms. PMID- 9395373 TI - Fibronectin-derived fragments as inducers of adhesion and chemotaxis of Entamoeba histolytica trophozoites. AB - Active migration of Entamoeba histolytica trophozoites through extracellular matrixes might play a role in host tissue destruction. Trophozoites degrade soluble fibronectin (FN) bound to their surface and adhere to substrate-bound FN, producing local degradation. FN proteolytic fragments were used to determine the nature of adhesion and motility-promoting domains within the protein. The 70-kDa fragment (amino-terminal end) promoted the highest adhesion, followed by the 120 kDa fragment, which contains the cell-binding domain. The 25-kDa fragment (carboxy-terminal end of the A chain) promoted half the adhesion, while two Hep II-binding fragments had no effect. The 70- and 120-kDa fragments also stimulated directed migration and chemokinesis. Intact FN and the 25-kDa fragment showed lower stimulation. The Hep II-binding fragments had no activity. Results support previous evidence for distinct cell-surface components as mediators of adhesion to FN and trophozoite motility and the potential importance of cell matrix recognition and degradation in their invasive behavior. PMID- 9395372 TI - Mutations in Plasmodium falciparum dihydrofolate reductase and dihydropteroate synthase and epidemiologic patterns of pyrimethamine-sulfadoxine use and resistance. AB - To assess the relationship between mutations in Plasmodium falciparum dihydrofolate reductase (DHFR) and dihydropteroate synthase (DHPS) and clinical pyrimethamine-sulfadoxine resistance, polymerase chain reaction surveys and analyses for new mutations were conducted in four countries with increasing levels of pyrimethamine-sulfadoxine resistance: Mali, Kenya, Malawi, and Bolivia. Prevalence of mutations at DHFR codon 108 and a new mutation at DHPS 540 correlated with increased pyrimethamine-sulfadoxine resistance (P < .05). Mutations at DHFR 51, DHFR 59, and DHPS 437 correlated with resistance without achieving statistical significance. Mutations at DHFR 164 and DHPS 581 were common in Bolivia, where pyrimethamine-sulfadoxine resistance is widespread, but absent in African sites. Two new DHFR mutations, a point mutation at codon 50 and an insert at codon 30, were found only in Bolivia. DHFR and DHPS mutations occur in a progressive, stepwise fashion. Identification of specific sets of mutations causing in vivo drug failure may lead to the development of molecular surveillance methods for pyrimethamine-sulfadoxine resistance. PMID- 9395374 TI - Review: JC virus infection of lymphocytes--revisited. AB - JC virus (JCV), the causative agent of the fatal human demyelinating disease progressive multifocal leukoencephalopathy (PML), is an opportunistic papovavirus that infects and destroys oligodendrocytes, the myelin-producing cells of the central nervous system. Since its isolation from the brain of a PML patient, JCV has long been classed as a neurotropic virus. Many studies, however, have demonstrated that JCV can infect various other cell types, including immune system cells. Moreover, several recent studies have focused specifically on lymphocytes as a target of JCV. This review chronicles the association of JCV with lymphocytes, including cell type localization, molecular regulation, and viral sequences, and discusses clinical implications of these findings. PMID- 9395375 TI - Hepatitis A virus infections in urban children--are preventive opportunities being missed? AB - To determine the prevalence of hepatitis A virus (HAV) infections in children in a large urban center, a point prevalence survey was conducted using a novel, ultrasensitive assay for HAV-specific IgG in saliva. A structured sample of 224 grade-six students (5.8% of grade registrants) was obtained from 23 schools throughout Vancouver. All students provided saliva samples adequate for testing. The anti-HAV prevalence rate was 7.1% (95% confidence interval, 4.1%-11.3%). Among 167 Canadian-born students, only 5 (3%) were positive, whereas among 57 students born elsewhere, 11 (19.3%) were positive (P < .001), with circumstances in the latter group supporting infection prior to emigration. No clustering of positive persons was evident. The cumulative risk of HAV infection in Canadian born children was low through age 11-12 years even in less affluent parts of the city, speaking against a need for routine use of HAV vaccine in this setting. PMID- 9395376 TI - High rate of hepatitis C virus clearance in hemodialysis patients after interferon-alpha therapy. AB - To gain insight into the long-term effect of interferon-alpha (IFN-alpha) therapy on hepatitis C virus (HCV) RNA-positive hemodialysis patients, 23 subjects were given 3 MU of IFN-alpha 3 times a week for 6 (n = 12) or 12 months (n = 11). They were followed for 19 months after cessation of therapy. Sustained serum HCV RNA clearance occurred in 42% of patients treated for 6 months and in 64% of those treated for 12 months. HCV was eradicated from 6 of 13 patients infected with HCV genotype 1b and from 2 of 6 patients also infected with hepatitis G virus. HCV RNA remained undetectable in both serum and a liver biopsy of 2 patients who were given cadaveric kidney transplants after IFN-alpha treatment. These data suggest that HCV RNA-positive dialysis patients can be considered for treatment while receiving dialysis, particularly those awaiting transplant. PMID- 9395377 TI - Investigation of human urine for genomic sequences of the primate polyomaviruses simian virus 40, BK virus, and JC virus. AB - Recent reports of the detection of simian virus 40 (SV40) nucleotide sequences in ependymomas, choroid plexus tumors, osteosarcomas, and mesotheliomas have raised the possibility that SV40, which naturally infects Asian macaques, is circulating among humans. This possibility was examined by performing polymerase chain reaction assays on urine samples of 166 homosexual men, 88 of them human immunodeficiency virus (HIV)-seropositive, for genomic sequences of SV40 as well as of human polyomaviruses BK virus (BKV) and JC virus (JCV). Tests with masked urine specimens spiked with SV40-transformed cells were included to monitor the SV40 assay. SV40, BKV, and JCV sequences were identified, respectively, in 0, 14%, and 34% of the urine specimens. JCV viruria was far more common (37%) than BKV viruria (5%) in HIV-seronegative persons. HIV infection and more severe immunosuppression were associated with a higher frequency of BKV viruria. In summary, SV40 viruria was not detected among homosexual men who shed human polyomaviruses at a high frequency. PMID- 9395378 TI - Plasma levels of monocyte chemoattractant protein-1 but not those of macrophage inhibitory protein-1alpha and RANTES correlate with virus load in human immunodeficiency virus infection. AB - Plasma levels of proinflammatory cytokines, cytokine inhibitors, and the beta chemokines RANTES, macrophage inhibitory protein (MIP)-1alpha, and monocyte chemoattractant protein (MCP)-1 were studied in relationship with virus load in 40 patients exhibiting plasma levels of HIV RNA ranging between undetectable and levels >10(6) copies/mL. Mean plasma levels of MCP-1 were increased in patients with high virus load compared with HIV-seropositive subjects with undetectable plasma viral RNA and healthy controls. MCP-1 levels were directly correlated with plasma levels of HIV RNA. No correlation was observed between virus load and plasma concentrations of MIP-1alpha and RANTES. The results suggest that low rates of viral replication in vivo are not dependent on increased production of the suppressive chemokines RANTES and MIP-1alpha. Since MCP-1 upregulates viral replication in vitro, the results may suggest a role for MCP-1 in triggering viral replication in HIV disease. PMID- 9395379 TI - An outbreak of foodborne illness caused by Escherichia coli O39:NM, an agent not fitting into the existing scheme for classifying diarrheogenic E. coli. AB - An outbreak of gastrointestinal illness with clinical and epidemiologic features of enterotoxigenic Escherichia coli (ETEC) occurred among patrons of a restaurant during April 1991. Illnesses among several groups of patrons were characterized by diarrhea (100%) and cramps (79%-88%) lasting a median of 3-5 days. Median incubation periods ranged from 50 to 56 h. A nonmotile strain of E. coli (E. coli O39), which was negative for heat-labile (LT) and heat-stable (STa, STb) ETEC toxins, was isolated only from ill patrons. This organism produced enteroaggregative E. coli heat-stable enterotoxin 1 and contained the enteropathogenic E. coli gene locus for enterocyte effacement; it did not display mannose-resistant adherence, but produced attaching and effacing lesions in the absence of mannose on cultured HEp-2 cells. E. coli that are not part of highly characterized but narrowly defined groups may be important causes of foodborne illness. PMID- 9395381 TI - Cytokine profiles in cerebrospinal fluid of human immunodeficiency virus-infected patients with cryptococcal meningitis: no leukocytosis despite high interleukin-8 levels. University of Zimbabwe Meningitis Group. AB - Cytokine levels were studied in the cerebrospinal fluid (CSF) of 16 adults with cryptococcal meningitis (CM). Low levels of tumor necrosis factor (TNF)-alpha and interferon-gamma, high levels of interleukin (IL)-1beta, IL-6, and IL-8, and the presence of IL-10 were documented. There were no significant differences in levels of TNF-alpha and interferon-gamma for CM and control patients. Mean CSF levels of IL-1beta (139.5 pg/mL), IL-6 (346 pg/mL), IL-8 (1160 pg/mL), and IL-10 (9.27 pg/mL) were significantly (P < .01) elevated in CM patients compared with levels in control patients. Despite the high CSF levels of IL-8, minimal leukocytosis was seen. Significant correlations between cryptococcal antigen titers and IL-10 levels (r = .8, P < .05), protein and cryptococcal antigen titer (r = .9, P < .05), and protein and IL-10 levels (r = .8, P < .05) were found. PMID- 9395380 TI - Presence of human immunodeficiency virus (HIV) type 1 subtype A infection in a New York community with high HIV prevalence: a sentinel site for monitoring HIV genetic diversity in North America. Centers for Disease Control and Prevention Bronx Lebanon HIV Serosurvey Team. AB - To determine whether US residents are infected with subtypes of human immunodeficiency virus (HIV) type 1 other than subtype B (Western), the predominant North American subtype with a unique GPGR genetic sequence in the V3 loop, viruses from 22 HIV-infected adults were serotyped and subtyped. Twenty patients had subtype B (Western), of whom 15 had serotype B (Western), 3 had serotype A/C, 1 had serotype B (Thai), and 1 had a nontypeable serotype. Two had subtype A, both serotype A/C. Both subtype A-infected patients, only 1 of whom had been outside the United States, reported sex with persons traveling abroad, suggesting possible acquisition in the United States. Because US residents are infected with non-subtype B (Western) strains, US surveillance for HIV-1 diversity is needed to elucidate subtype-specific transmission patterns and pathogenesis and to guide evaluation and development of HIV diagnostic tests and vaccines. PMID- 9395382 TI - Staphylococcus aureus stimulates urokinase-type plasminogen activator expression by bovine mammary cells. AB - Staphylococcus aureus commonly causes bovine mastitis, but bovine strains, unlike human isolates of S. aureus, do not produce the bacterial plasminogen activator, staphylokinase. By use of bovine mammary epithelial and myoepithelial cell lines, it was found that bovine S. aureus M60 and its culture filtrates induce a 3- to 10-fold increase in urokinase-type plasminogen activator activity in mammary cell conditioned media and cellular lysates. Furthermore, transcytosis of S. aureus M60 across a mammary epithelial cell monolayer was significantly enhanced by the addition of bovine plasminogen and inhibited by aprotinin. These findings provide evidence that S. aureus M60 can trigger superactivation of host plasminogen activator production and may then utilize the plasminogen activator-plasmin(ogen) system to facilitate tissue invasion without producing staphylokinase. PMID- 9395383 TI - Risk for gastric lymphoma in persons with CagA+ and CagA- Helicobacter pylori infection. AB - Infection with Helicobacter pylori increases the risk for gastric non-Hodgkin's lymphoma (GNHL). Strains that express CagA protein are thought to be particularly virulent. It was determined whether CagA+ H. pylori infection increased the risk for GNHL more than CagA infection. Thirty-two cases and 130 controls previously tested for H. pylori antibodies were tested for CagA antibodies by ELISA. The risk for GNHL was compared among CagA+, CagA-, and uninfected persons by use of conditional logistic regression. CagA+ subjects had 8.2 times the risk for GNHL than uninfected persons (95% confidence interval [CI], 2.5-26.7). CagA- subjects had 4.4 times the risk for GNHL than uninfected persons (95% CI, 1.2-16.5). Among infected subjects only, CagA+ infection was not associated with significantly increased risk for GNHL when compared with CagA- infection (odds ratio, 2.1; 95% CI, 0.8-5.4). This study does not support a major role for CagA in lymphomagenesis. PMID- 9395384 TI - Lymphocyte response to tetanus toxoid among Indonesian men immunized with tetanus diphtheria during extended chloroquine or primaquine prophylaxis. AB - Immune suppression, a potential side effect of long-term chemoprophylaxis, was evaluated as part of a randomized, placebo-controlled trial that compared daily primaquine against weekly chloroquine for malaria prevention. In the last month of the year-long trial, baseline in vitro lymphoproliferative responses to tetanus toxoid were measured, and a tetanus-diphtheria (Td) immunization was administered. Proliferative responses to tetanus toxoid in each Td-immunized group increased significantly over pre-Td baselines and those of the unvaccinated control. Highest initial responses were measured in the primaquine group. The proportion of responders and the magnitude of proliferation was consistently low in the chloroquine group, and end point responses in this group were significantly below those of the placebo. These results suggest that the development and duration of the cellular response to tetanus immunization was impaired by long-term weekly chloroquine prophylaxis, while daily primaquine prophylaxis over the same time period had no inhibitory effect. PMID- 9395385 TI - Gammadelta T cells are not essential for control of cutaneous Leishmania major infection in genetically resistant C57BL/6 mice. AB - As gammadelta T cells are believed to be involved in host defense against Leishmania major, the role of gammadelta T cells in immunity against this parasite was investigated. The growth of L. major was measured in alphabeta (T cell receptor [TCR] alpha -/-) and gammadelta (TCR delta -/-) TCR-deficient C57BL/6 mice and compared with growth in control (C57BL/6) mice. While TCR alpha /- mice developed nonhealing lesions containing large numbers of parasites following L. major infection, TCR delta -/- and C57BL/6 mice effectively controlled the infection. Following in vitro stimulation, lymph node cells from C57BL/6 mice produced significantly more interferon (IFN)-gamma than those from TCR delta -/- mice during early and late phases of infection; however, both produced similar levels of IFN-gamma at postinfection week 6. Culture supernatants from both TCR delta -/- and C57BL/6 mice contained interleukin-4, at postinfection week 2 only. These results indicate that gammadelta CD3 T cells are not essential for mediating protection against cutaneous L. major infection in C57BL/6 mice. PMID- 9395386 TI - Construction crew discovers grave of "Tiny Tim". PMID- 9395387 TI - An inverse compton process for the excess diffuse EUV emission from the virgo and coma galaxy clusters AB - The excess extreme-ultraviolet (EUV) emission detected in the Virgo and Coma clusters is explained by inverse Compton scattering of cosmic microwave background photons, which are scattered by the relativistic electrons that account for the extended radio synchrotron emission of these clusters. The lower limits of the average magnetic fields of these clusters estimated from the EUV excess are close to the equipartition magnetic fields derived from radio observations, indicating that the electron energies and magnetic field energies might be close to equipartition. The excess emission suggests energy reservoirs of approximately 10(61) and approximately 10(60) ergs for the Coma and Virgo clusters, respectively. PMID- 9395388 TI - A pulsar, the heliosphere, and pioneer 10: probable mimicking of a planet of PSR B1257+12 by solar rotation AB - Doppler data generated with the Pioneer 10 spacecraft's radio carrier wave between 1987 and 1995 show a 25.3-day periodicity which is related to the solar rotation. The timing data of the pulsar PSR B1257+12 also show a periodicity of 25.34 days, which has been explained as a signature of the pulsar's barycentric motion in response to the existence of a small moon-like object. However, because PSR B1257+12 is located close to the ecliptic and because the timing variations are in the range of microseconds, it is likely that the pulsar signal is affected by the same mechanism acting on the Pioneer 10 Doppler data. Hence, the hypothesized inner planet around PSR B1257+12 is probably an artifact of the heliosphere. PMID- 9395389 TI - Radar detection of the nucleus and coma of comet hyakutake AB - Radar observations of comet Hyakutake (C/1996 B2) made at the Goldstone Deep Space Communications Complex in California have detected echoes from the nucleus and from large grains in the inner coma. The nucleus of this bright comet was estimated to be only 2 to 3 kilometers in diameter. Models of the coma echo indicate backscatter from porous, centimeter-size grains ejected anisotropically at velocities of tens of meters per second. The radar observations suggest that a comet's activity may be a poor indicator of its size and provide evidence that large grains constitute an important component of the mass loss from a typical active comet. PMID- 9395390 TI - Spontaneous formation of macroscopic chiral domains in a fluid smectic phase of achiral molecules AB - A smectic liquid-crystal phase made from achiral molecules with bent cores was found to have fluid layers that exhibit two spontaneous symmetry-breaking instabilities: polar molecular orientational ordering about the layer normal and molecular tilt. These instabilities combine to form a chiral layer structure with a handedness that depends on the sign of the tilt. The bulk states are either antiferroelectric-racemic, with the layer polar direction and handedness alternating in sign from layer to layer, or antiferroelectric-chiral, which is of uniform layer handedness. Both states exhibit an electric field-induced transition from antiferroelectric to ferroelectric. PMID- 9395391 TI - "New view" of protein folding reconciled with the old through multiple unfolding simulations. AB - Twenty-four molecular dynamics trajectories of chymotrypsin inhibitor 2 provide a direct demonstration of the diversity of unfolding pathways. Comparison with experiments suggests that the transition state region for folding and unfolding occurs early with only 25 percent of the native contacts and that the root-mean square deviations between contributing structures can be as large as 15 angstroms. Nevertheless, a statistically preferred unfolding pathway emerges from the simulations; disruption of tertiary interactions between the helix and a two stranded portion of the beta sheet is the primary unfolding event. The results suggest a synthesis of the "new" and the classical view of protein folding with a preferred pathway on a funnel-like average energy surface. PMID- 9395392 TI - Atomic and macroscopic reaction rates of a surface-catalyzed reaction AB - The catalytic oxidation of carbon monoxide (CO) on a platinum (111) surface was studied by scanning tunneling microscopy. The adsorbed oxygen atoms and CO molecules were imaged with atomic resolution, and their reactions to carbon dioxide (CO2) were monitored as functions of time. The results allowed the formulation of a rate law that takes the distribution of the reactants in separate domains into account. From temperature-dependent measurements, the kinetic parameters were obtained. Their values agree well with data from macroscopic measurements. In this way, a kinetic description of a chemical reaction was achieved that is based solely on the statistics of the underlying atomic processes. PMID- 9395393 TI - Growth of SiO2 at room temperature with the use of catalyzed sequential half reactions AB - Films of silicon dioxide (SiO2) were deposited at room temperature by means of catalyzed binary reaction sequence chemistry. The binary reaction SiCl4 + 2H2O - > SiO2 + 4HCl was separated into SiCl4 and H2O half-reactions, and the half reactions were then performed in an ABAB ellipsis sequence and catalyzed with pyridine. The pyridine catalyst lowered the deposition temperature from >600 to 300 kelvin and reduced the reactant flux required for complete reactions from approximately 10(9) to approximately 10(4) Langmuirs. Growth rates of approximately 2.1 angstroms per AB reaction cycle were obtained at room temperature for reactant pressures of 15 millitorr and 60-second exposure times with 200 millitorr of pyridine. This catalytic technique may be general and should facilitate the chemical vapor deposition of other oxide and nitride materials. PMID- 9395394 TI - Cleavage of the BMP-4 antagonist chordin by zebrafish tolloid. AB - Dorsoventral patterning of vertebrate and Drosophila embryos requires bone morphogenetic proteins (BMPs) and antagonists of BMP activity. The Drosophila gene tolloid encodes a metalloprotease similar to BMP-1 that interacts genetically with decapentaplegic, the Drosophila homolog of vertebrate BMP-2/4. Zebrafish embryos overexpressing a zebrafish homolog of tolloid were shown to resemble loss-of-function mutations in chordino, the zebrafish homolog of the Xenopus BMP-4 antagonist Chordin. Furthermore, Chordin was degraded by COS cells expressing Tolloid. These data suggest that Tolloid antagonizes Chordin activity by proteolytically cleaving Chordin. A conserved function for zebrafish and Drosophila Tolloid during embryogenesis is proposed. PMID- 9395395 TI - Role of inositol 1,4,5-trisphosphate receptor in ventral signaling in Xenopus embryos. AB - The inositol 1,4,5-trisphosphate (IP3) receptor is a calcium ion channel involved in the release of free Ca2+ from intracellular stores. For analysis of the role of IP3-induced Ca2+ release (IICR) on patterning of the embryonic body, monoclonal antibodies that inhibit IICR were produced. Injection of these blocking antibodies into the ventral part of early Xenopus embryos induced modest dorsal differentiation. A close correlation between IICR blocking potencies and ectopic dorsal axis induction frequency suggests that an active IP3-Ca2+ signal may participate in the modulation of ventral differentiation. PMID- 9395396 TI - Crystal structure of the adenylyl cyclase activator Gsalpha. AB - The crystal structure of Gsalpha, the heterotrimeric G protein alpha subunit that stimulates adenylyl cyclase, was determined at 2.5 A in a complex with guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS). Gsalpha is the prototypic member of a family of GTP-binding proteins that regulate the activities of effectors in a hormone-dependent manner. Comparison of the structure of Gsalpha.GTPgammaS with that of Gialpha.GTPgammaS suggests that their effector specificity is primarily dictated by the shape of the binding surface formed by the switch II helix and the alpha3-beta5 loop, despite the high sequence homology of these elements. In contrast, sequence divergence explains the inability of regulators of G protein signaling to stimulate the GTPase activity of Gsalpha. The betagamma binding surface of Gsalpha is largely conserved in sequence and structure to that of Gialpha, whereas differences in the surface formed by the carboxyl-terminal helix and the alpha4-beta6 loop may mediate receptor specificity. PMID- 9395397 TI - Mediation of Sonic hedgehog-induced expression of COUP-TFII by a protein phosphatase. AB - A Sonic hedgehog (Shh) response element was identified in the chicken ovalbumin upstream promoter-transcription factor II (COUP-TFII) promoter that binds to a factor distinct from Gli, a gene known to mediate Shh signaling. Although this binding activity is specifically stimulated by Shh-N (amino-terminal signaling domain), it can also be unmasked with protein phosphatase treatment in the mouse cell line P19, and induction by Shh-N can be blocked by phosphatase inhibitors. Thus, Shh-N signaling may result in dephosphorylation of a target factor that is required for activation of COUP-TFII-, Islet1-, and Gli response element dependent gene expression. This finding identifies another step in the Shh-N signaling pathway. PMID- 9395398 TI - Spike synchronization and rate modulation differentially involved in motor cortical function. AB - It is now commonly accepted that planning and execution of movements are based on distributed processing by neuronal populations in motor cortical areas. It is less clear, though, how these populations organize dynamically to cope with the momentary computational demands. Simultaneously recorded activities of neurons in the primary motor cortex of monkeys during performance of a delayed-pointing task exhibited context-dependent, rapid changes in the patterns of coincident action potentials. Accurate spike synchronization occurred in relation to external events (stimuli, movements) and was commonly accompanied by discharge rate modulations but without precise time locking of the spikes to these external events. Spike synchronization also occurred in relation to purely internal events (stimulus expectancy), where firing rate modulations were distinctly absent. These findings indicate that internally generated synchronization of individual spike discharges may subserve the cortical organization of cognitive motor processes. PMID- 9395399 TI - Protein disulfide isomerase as a regulator of chloroplast translational activation. AB - Light-regulated translation of chloroplast messenger RNAs (mRNAs) requires trans acting factors that interact with the 5' untranslated region (UTR) of these mRNAs. Chloroplast polyadenylate-binding protein (cPABP) specifically binds to the 5'-UTR of the psbA mRNA and is essential for translation of this mRNA. A protein disulfide isomerase that is localized to the chloroplast and copurifies with cPABP was shown to modulate the binding of cPABP to the 5'-UTR of the psbA mRNA by reversibly changing the redox status of cPABP through redox potential or adenosine 5'-diphosphate-dependent phosphorylation. This mechanism allows for a simple reversible switch regulating gene expression in the chloroplast. PMID- 9395400 TI - Sequence-specific and phosphorylation-dependent proline isomerization: a potential mitotic regulatory mechanism. AB - Pin1 is an essential and conserved mitotic peptidyl-prolyl isomerase (PPIase) that is distinct from members of two other families of conventional PPIases, cyclophilins and FKBPs (FK-506 binding proteins). In response to their phosphorylation during mitosis, Pin1 binds and regulates members of a highly conserved set of proteins that overlaps with antigens recognized by the mitosis specific monoclonal antibody MPM-2. Pin1 is here shown to be a phosphorylation dependent PPIase that specifically recognizes the phosphoserine-proline or phosphothreonine-proline bonds present in mitotic phosphoproteins. Both Pin1 and MPM-2 selected similar phosphorylated serine-proline-containing peptides, providing the basis for the specific interaction between Pin1 and MPM-2 antigens. Pin1 preferentially isomerized proline residues preceded by phosphorylated serine or threonine with up to 1300-fold selectivity compared with unphosphorylated peptides. Pin1 may thus regulate mitotic progression by catalyzing sequence specific and phosphorylation-dependent proline isomerization. PMID- 9395401 TI - Mechanism of transcription through the nucleosome by eukaryotic RNA polymerase. AB - Nucleosomes, the nucleohistone subunits of chromatin, are present on transcribed eukaryotic genes but do not prevent transcription. It is shown here that the large yeast RNA polymerase III transcribes through a single nucleosome. This takes place through a direct internal nucleosome transfer in which histones never leave the DNA template. During this process, the polymerase pauses with a pronounced periodicity of 10 to 11 base pairs, which is consistent with restricted rotation in the DNA loop formed during transfer. Transcription through nucleosomes by the eukaryotic enzyme and by much smaller prokaryotic RNA polymerases thus shares many features, reflecting an important property of nucleosomes. PMID- 9395402 TI - NDR1, a pathogen-induced component required for Arabidopsis disease resistance. AB - Plant disease resistance (R) genes confer an ability to resist infection by pathogens expressing specific corresponding avirulence genes. In Arabidopsis thaliana, resistance to both bacterial and fungal pathogens, mediated by several R gene products, requires the NDR1 gene. Positional cloning was used to isolate NDR1, which encodes a 660-base pair open reading frame. The predicted 219-amino acid sequence suggests that NDR1 may be associated with a membrane. NDR1 expression is induced in response to pathogen challenge and may function to integrate various pathogen recognition signals. PMID- 9395403 TI - Conversion of Bcl-2 to a Bax-like death effector by caspases. AB - Caspases are a family of cysteine proteases implicated in the biochemical and morphological changes that occur during apoptosis (programmed cell death). The loop domain of Bcl-2 is cleaved at Asp34 by caspase-3 (CPP32) in vitro, in cells overexpressing caspase-3, and after induction of apoptosis by Fas ligation and interleukin-3 withdrawal. The carboxyl-terminal Bcl-2 cleavage product triggered cell death and accelerated Sindbis virus-induced apoptosis, which was dependent on the BH3 homology and transmembrane domains of Bcl-2. Inhibitor studies indicated that cleavage of Bcl-2 may further activate downstream caspases and contribute to amplification of the caspase cascade. Cleavage-resistant mutants of Bcl-2 had increased protection from interleukin-3 withdrawal and Sindbis virus induced apoptosis. Thus, cleavage of Bcl-2 by caspases may ensure the inevitability of cell death. PMID- 9395404 TI - A dual effect of 1-methyl-4-phenylpyridinium (MPP+)-analogs on the respiratory chain of bovine heart mitochondria. AB - We examined effects of several compounds, structurally related to 1-methyl-4 phenylpyridinium (MPP+), on the NADH-dependent respiration of bovine heart submitochondrial particles. 1-Methyl-4-(3 '-trimethylammoniophenyl)pyridinium (analog 8) as well as MPP+ completely inhibited O2 consumption, reduction of ubiquinone-10, and reduction of cytochrome b in a dose-dependent manner. The production of superoxide (O2-) induced by MPP+ or analog 8 was to the same extent as that by rotenone, an inhibitor of complex I of the mitochondrial respiratory chain. Rotenone had no additive effect on the maximal production of O2- induced by MPP+ or analog 8, suggesting that the production was mediated by the same way as rotenone. 1-Methyl-4-(4'-nitrophenyl) pyridinium (analog 1) induced about 20 fold more production of O2 than MPP+ and the production was additively increased by rotenone. Analog 1 only partially inhibited rotenone-sensitive O2 consumption. Paraquat induced the production of O2- as much as analog 1. Paraquat, however, did not inhibit rotenone-sensitive O2 consumption or reduction of cytochrome b. These results suggest that MPP+ and its analogs interact with the mitochondrial respiratory chain at two sites, the substrate side of the rotenone-binding site and the rotenone-binding site. The analogs may be reduced to produce O2- at the former site and inhibit the respiratory chain at the latter site. PMID- 9395405 TI - Evolutionary motif and its biological and structural significance. AB - We developed a method for multiple alignment of protein sequences. The main feature of this method is that it takes the evolutionary relationships of the proteins in question into account repeatedly for execution, until the relationships and alignment results are in agreement. We then applied this method to the data of the international DNA sequence databases, which are the most comprehensive and updated DNA databases in the world, in order to estimate the "evolutionary motif" by extensive use of a supercomputer. Though a few problems needed to be solved, we could estimate the length of the motifs in the range of 20 to 200 amino acids, with about 60 the most frequent length. We then discussed their biological and structural significance. We believe that we are now in a position to analyze DNA and protein not only in vivo and in vitro but also in silico. PMID- 9395406 TI - Genome plasticity as a paradigm of eubacteria evolution. AB - To test the hypotheses that eubacterial genomes leave evolutionarily stable structures and that the variety of genome size is brought about through genome doubling during evolution, the genome structures of Haemophilus influenzae, Mycoplasma genitalium, Escherichia coli, and Bacillus subtilis were compared using the DNA sequences of the entire genome or substantial portions of genome. In these comparisons, the locations of orthologous genes were examined among different genomes. Using orthologous genes for the comparisons guaranteed that differences revealed in physical location would reflect changes in genome structure after speciation. We found that dynamic rearrangements have so frequently occurred in eubacterial genomes as to break operon structures during evolution, even after the relatively recent divergence between E. coli and H. influenzae. Interestingly, in such eubacterial genomes of high plasticity, we could find several highly conservative regions with the longest conserved region comprising the S10, spc, and alpha operons. This suggests that such exceptional conservative regions have undergone strong structural constraints during evolution. PMID- 9395407 TI - Requirement for EGF receptor signalling in neural recruitment during formation of Drosophila chordotonal sense organ clusters. AB - BACKGROUND: Drosophila proneural genes act in the process of selecting neural precursors from undifferentiated ectoderm. The proneural gene atonal is required for the development of precursors of both chordotonal organs (stretch receptors) and photoreceptors. Although these types of sensory element are dissimilar in structure and function, they both occur as organized arrays of neurons. Previous studies have shown that clustering of photoreceptors involves local recruitment, and that signalling by the Drosophila epidermal growth factor receptor (DER) pathway is involved in the recruitment process. We present evidence that a similar mechanism is required for the clustering of embryonic chordotonal organs. RESULTS: We have examined the expression patterns of atonal and genes of the DER pathway in wild-type and mutant backgrounds. Expression of atonal was restricted to a subset of the atonal-requiring chordotonal precursors, which we call founder precursors. The remaining precursors required DER signalling for their selection. Signalling by the founder precursors was initiated by atonal activating, directly or indirectly, rhomboid expression in these cells. Signalling by these founder precursors then provoked a response in the surrounding ectodermal cells, as shown by the activation of expression of the DER target genes pointed and argos. The signal and response then led to recruitment of some of the ectodermal cells to the chordotonal precursor cell fate. DER hyperactivation by misexpression of rhomboid resulted in excessive chordotonal precursor recruitment. CONCLUSIONS: Increased numbers of chordotonal precursors are recruited by homeogenetic induction involving signalling via DER from founder precursors to surrounding ectodermal cells. We suggest that the reason chordotonal organs and photoreceptors share a requirement for the proneural gene atonal is that this gene activates a common pathway leading to neural aggregation. PMID- 9395409 TI - Movement of nuclei along microtubules in Xenopus egg extracts. AB - Microtubules are implicated in the movement and positioning of nuclei in many cell types. Nuclei can be moved by forces acting on microtubules nucleated at the spindle pole body, as in fungi [1], or microtubules nucleated at the centrosome, as during migration of the male (sperm) pronucleus towards the centre of the zygote after fertilization [2] [3] [4]. The dramatic movements of the female pronucleus towards the male pronucleus potentially involve another mechanism: movement along the microtubules of the sperm aster towards their slower growing, attached or 'minus' ends [3] [5]. Here, we have reconstituted this last type of nuclear movement in vitro. Synthetic nuclei assembled in cytoplasmic extracts made from interphase Xenopus eggs move along microtubules towards their minus ends. We provide strong experimental evidence that cytoplasmic dynein is the motor for nuclear movement in this in vitro system, and discuss our results in terms of current knowledge of motility of the endoplasmic reticulum. PMID- 9395408 TI - Phospholipase D2, a distinct phospholipase D isoform with novel regulatory properties that provokes cytoskeletal reorganization. AB - BACKGROUND: Activation of phospholipase D (PLD) is an important but poorly understood component of receptor-mediated signal transduction responses and regulated secretion. We recently reported the cloning of the human gene encoding PLD1; this enzyme has low basal activity and is activated by protein kinase C and the small GTP-binding proteins, ADP-ribosylation factor (ARF), Rho, Rac and Cdc42. Biochemical and cell biological studies suggest, however, that additional and distinct PLD activities exist in cells, so a search was carried out for novel mammalian genes related to PLD1. RESULTS: We have cloned the gene for a second PLD family member and characterized the protein product, which appears to be regulated differently from PLD1: PLD2 is constitutively active and may be modulated in vivo by inhibition. Unexpectedly, PLD2 localizes primarily to the plasma membrane, in contrast to PLD1 which localizes solely to peri-nuclear regions (the endoplasmic reticulum, Golgi apparatus and late endosomes), where PLD activity has been shown to promote ARF-mediated coated-vesicle formation. PLD2 provokes cortical reorganization and undergoes redistribution in serum stimulated cells, suggesting that it may have a role in signal-induced cytoskeletal regulation and/or endocytosis. CONCLUSIONS: PLD2 is a newly identified mammalian PLD isoform with novel regulatory properties. Our findings suggest that regulated secretion and morphological reorganization, the two most frequently proposed biological roles for PLD, are likely to be effected separately by PLD1 and PLD2. PMID- 9395411 TI - [Why not a meeting place for the readers of the L[kartidningen?]. PMID- 9395410 TI - [Impact of control measures on the course of an outbreak of methicillin-resistant Staphylococcus aureus]. AB - BACKGROUND: Outbreaks of nosocomial infection by methicillin resistent Staphylococcus aureus (MRSA) are a problem in many hospitals with the control measures to be adopted being controversial. An outbreak of MRSA in a 550-bed university hospital is herein described and the impact of the adopted control measures on the evolution of the epidemic in the general hospitalization area (GHA) was analyzed. PATIENTS AND METHODS: The adopted control measures in the GHA were: microbiologic surveillance, cutaneous isolation measures, treatment of nasal carrier, and the early discharge of the cases. Hand washing was reinforced and a study of carriers was carried out on detection of sporadic cases (not related to the ICU). A molecular study of 70 strains of MRSA was performed with analysis of total plasmids, plasmid restriction pattern and chromosomic DNA analysis by pulsed field gel electrophoresis (PFGE). RESULTS: From December 1990 to December 1993, 273 cases of MRSA were reported. One hundred seventy-two cases originated in the ICU and 101 cases in the GHA (sporadic cases). The incidence of MRSA in 1991-1993 was 13.6, 14.3, and 6.6% in the ICU and 0.17, 0.36, and 0.15% in the GHA, respectively. Molecular study of MRSA isolates (1991 and 1992) demonstrated two plasmid and two chromosomic patterns. The latter had a similarity coefficient > 0.90, probably belonging to the same "clone". CONCLUSIONS: Despite the control measures adopted in the GHA the outbreak of MRSA originated in the ICU thereafter extending to the GHA. The rates of colonization detected, however, remained stable during the 3 years studied. On the other hand, the observation of a single "clone", responsible for the epidemic, suggest that most of the sporadic cases were autoctonous and due to failure in fulfillment of the established norms. PMID- 9395412 TI - [Claves ilustradas para las subfamilias, tribus y generos de esfecidos neotropicales (Apoidea: Sphecidae) [Illustrated clues for subfamilies, tribes, and genera of neotropical Spheciade (Apoidea:Sphecidae)]. AB - 141 genera of Sphecidae, representing 1,628 species, are known from the Neotropical Region. Illustrated keys to genera, tribes, and subfamilies are presented in Spanish and English. These have been modified and updated from those in Bohart & Menke's 1976 book, Sphecid Wasps of the World. The validity of a few genera recognized by Bohart & Menke is now in question and the keys are annotated to alert users to these problems. A list of neotropical genera and higher taxa is included. Names in the list are appended with significant literature published since 1976. The history and current status of subfamilies are reviewed. Ten subfamilies are recognized. Family characters and biology are summarized. Morphological terms are illustrated and a glossary provided. PMID- 9395413 TI - [Directly addressing suicidal thoughts in crisis intervention. Interview by Dr. rer. nat. Anita Schweiger]. PMID- 9395414 TI - [Infectious endocarditis at the National Institute of Cardiology "Ignacio Chavez". Five year's experience (1990-1994)]. AB - BACKGROUND: Infective endocarditis (IE) is one of the most life threatening infections in both medical and surgical practices. In the last few years, changes in its epidemiology, diagnostic methods and therapeutical trends have appeared. We analysed our experience in the diagnosis and treatment of IE. METHODS: The clinical records of patients admitted to our hospital with definitive (Group I) and highly probable (Group II) diagnosis of IE, during a period of five years (1990-1994), were retrospectively reviewed. Age, sex, clinical features, risk, factors, echocardiographic abnormalities, microbiologic and surgical findings, as well as mortality were recorded. In addition, an evaluation was made of the accuracy of the diagnostic criteria proposed by Von Reyn versus those brought forward by Duke University. RESULTS: One hundred thirty one patients were included, 99 in Group I and 32 in Group II. The mean age was 35 years. Native valve endocarditis was present in 88 patients and prosthetic valve endocarditis in 43 patients. Streptococcus sp. (48%) was the most frequently causative german and 16.7% of cases were culture negative. The sensitivity of transesophageal echocardiography was higher than transthoracic echocardiography in the diagnosis of both vegetations (76% vs 55%) and abscesses (30% vs 16.5%), (p < 0.05). Vegetations (95%) were the most frequent surgical finding followed by abscesses (23%). Inpatient mortality was 22% in Group I and 45% in Group II (p < 0.05). The sensitivity of Von Reyn's diagnostic criteria and that of Duke's University group was 49% and 85.8% (p < 0.05). Mean follow up was 531 days. Two patients had a new event of IE and no outpatient deaths were recorded. CONCLUSION: IE is a medical and surgical emergency. Because of the high mortality rate, in the medically treated group, surgery should be considered in all cases as early as possible in the course of the disease. PMID- 9395415 TI - [[HPLC-determination of tamoxifen dihydrogen citrate with the base selective column UltraSep ES Pharm RP8 in tablets]. PMID- 9395416 TI - [The causes of perioperative mortality. A trial of the German "CEPOD study."]. AB - We performed a prospective multi-center study in order to determine the causes of 30-day perioperative mortality. METHODS: In accordance with the CEPOD-Study and with the kind permission of Dr. N. Lunn, we forwarded two different questionnaires to 135 hospitals. One questionnaire was to be answered by the anaesthetist and the other one by the surgeon involved in cases o perioperative death within the first 30 days after the operation. 12 out of 135 addressed hospitals agreed to participate in the study. These included four small hospitals, six medical centres of medium capacity (about 500 beds) and two University hospitals. In order to obtain an exact description of the events leading to perioperative death, the questionnaires consisted of approximately 60 questions for the collection of demographic data and the surgical as well as anaesthesiological perioperative management. RESULTS: From 1989 to 1993 more than 300 cases of perioperative death were reported. Only 200 cases could be analyzed due to incompletely answered or unreturned questionnaires. The mean risk classification (ASA) was 3.46, mean age 74.6 years. Approximately 40 percent of deaths occurred in patients older than 80 years. More than 80 percent of patients had at least one pre-existing cardiovascular disease with prevalence of 41% for pulmonary and gastrointestinal diseases. In the majority of cases abdominal operations were performed, followed by hip-surgery and surgery of the aorta. In 86% of the cases, the surgeon was experienced and had performed the respective operation more than 20 times. In 38.2% an anaesthetist in training was responsible for anaesthesia, but only 11.6% were without supervision of a specialist anaesthetist. The majority of patients received general anaesthesia (78%) and 8.5% had a combination of EDA and general anaesthesia. Regional anaesthesia was performed in 12.5%, local anaesthesia in only 1%. The average blood loss was approximately 1.600 ml (with a very wide range) and 42.5% of the patients needed a transfusion of blood components, primarily in the form of packed red blood cells. Seventeen serious incidents occurred intraoperatively, including three "exitus in tabula". Four patients died shortly after the operation in the ICU, the other ten incidents were managed in the operating room. In 11 of 17 incidents the patients suffered a cardiac arrest; nine patients were resuscitated. Two patients were not resuscitated in view of pre-existing diseases and inoperability. All of the hospitals had an ICU for postoperative care, but two of the smaller hospitals had no recovery rooms. In 22 cases of emergency operations, there was a delay due to a lack of personnel or to logistic problems. In five of these cases, the delay was described as a possible cofactor of perioperative mortality. The most frequent causes of perioperative death were myocardial failure (33.7%) and multi-organ-failure (19.2%), followed by respiratory insufficiency (13%) and septic shock in 9.3%. A necropsy was carried out in only 28 of 200 perioperative deaths (14%); 13% of the cases were discussed in a surgical and only 2.5% in an anaesthesiological mortality-conference. In 9 out of 12 hospitals no mortality-conferences were held. All surgeons and anaesthetists were asked for self-assessment on the basis of an analog scale ranging from 0 and 10 points. The average score was 8.52 points (surgical management) and 9.36 points (anaesthesiological management respectively), which is not always in correspondence with the information provided in the questionnaires. CONCLUSIONS: In order to further reduce perioperative mortality in critically ill patients, every hospital should aim to optimize the structure of the surgical and anaesthesiological departments. A delay due to logistical or personnel problems may be a co-factor in perioperative mortality. Recovery rooms with experienced personnel should be the standard in postoperative anaesthesiological care. (ABSTRACT TRUNCATED) PMID- 9395417 TI - [Institutionalization of community programs: review of theoretical models and proposal of a model-]. AB - Community-based health promotion programs to change lifestyle habits must remain in their host organizations for extended periods of time in order to have impact. Their effectiveness can be closely linked to their long term viability or institutionalization. To remain viable, these programs must survive beyond initial investment and support by external organizations. However, some programs disappear when external investment is withdrawn. This can be costly and in addition can generate resistance to the implementation of other health promotion programs in the future. Recently, interest in the processes involved in the institutionalization of these programs has increased. Based on 28 publications, this article reviews selected conceptual models that highlight environmental, organizational, community and marketing-related, variables possibly related to the institutionalization process. A new model is proposed to link these diverse models according to: characteristics of the program, characteristics of the host organization, characteristics related to the adoption, implementation and incorporation of the program, and finally characteristics related to the fit (mutual adjustment) between the host and the program. PMID- 9395418 TI - [[Brucellar orchiepididymitis with abscess]. AB - OBJECTIVE: To emphasize the need to consider Brucella infection in patients presenting with acute scrotum of a possible inflammatory etiology, in endemic areas, as in some Spanish regions. The abscess forming type, although rare, should be considered. One such case is described herein and the literature briefly reviewed. METHODS: A male patient consulted for subacute inflammation and left testicular pain. He had systemic brucellosis four months earlier that had completely remitted following specific therapy. The patient had a physical examination, analytical, blood and urine analyses, specific serological tests and testicular ultrasound evaluation. RESULTS: Physical examination disclosed left testicular pain and inflammation suggesting epididimo-orchitis. The laboratory findings were normal except for high titles against Brucella. Ultrasound disclosed diffuse enlargement of the left testis with several well-defined hypoechoic areas. The foregoing data suggested abscess forming orchitis, although a neoplasm could not be discarded. Empirical antibiotic therapy was instituted initially and specific therapy for Brucella was administered subsequently, but unilateral orchidectomy was decided because of the poor response to therapy. Histopathological analysis of the surgical specimen disclosed acute abscess forming epididimo-orchitis with multifocal chronic granulomatous involvement. CONCLUSION: Brucella epididimo-orchitis must be considered when making the differential diagnosis in acute inflammatory scrotum, particularly in endemic areas, even in the absence of suggestive clinical and/or US findings. Necrotizing orchitis is a rare form of Brucella infection which must be distinguished from necrotizing involvement arising from other pathogens (TB or Salmonella). Above all, this condition must be distinguished from a tumor. PMID- 9395419 TI - [Does breast feeding compromise the efficacy of sero-vaccination of infants borne to mothers who are chronic carriers of hepatitis B antigens]. PMID- 9395420 TI - [Are the products of CD44 exons v5 and v6 markers for metastasis of laryngeal carcinomas?]. AB - BACKGROUND: The transmembrane glycoprotein CD44 is referred to by many names, which are related to the polymorphism of this molecule. There are at least 10 different versions of the CD44 molecule. This polymorphism results from the insertion of extra domains into the extracellular part of the molecule and from different glycolization. These extra domains are coded by variable exons in the gene of CD44, which can be alternatively spliced. Some authors have postulated a link between expression of whole CD44 or some special molecule versions (often with the product of exon v6) on carcinoma cells and the potential of metastatic spread. The aim of our investigation was to look for this connection in larynx carcinomas. METHOD: We have tested 28 larynx carcinomas without metastases, 26 with metastases, and 20 lymph node metastases from larynx carcinomas with antibodies against the products of exon v5 and v6 in immunohistochemical studies of paraffin sections. RESULTS: In all cases we observed nearly the same staining intensity of exon v5 and v6 products. There was no significant difference between carcinomas with and without metastases or the lymph node metastases. However, a strong difference of reaction was found between the carcinoma cells of the outer proliferative tumor areas and the inner tumor areas, which were cornified in parts. Whereas the first mentioned cells generally stained very intensively, the latter showed only a slight reaction or none at all. CONCLUSIONS: In our opinion, CD44 v5 and v6 appear to be valuable markers of proliferation although we could not establish a strong connection to metastatic behavior. PMID- 9395421 TI - [Chronic fatigue syndrome: study of the clinical course of 28 cases]. AB - BACKGROUND: Chronic fatigue syndrome (CFS) is an entity of unknown etiopathogenesis without specific markers. The diagnosis is based on clinical criteria. There are few studies evaluating the natural evolution and prognosis related factors in CFS. OBJECTIVES: a) to evaluate the outcome of patients suffering from CFS, and b) to detect predictive factors associated with a better prognosis. MATERIAL AND METHODS: Clinical records of all patients diagnosed of CFS between January 1986 and December 1992 were retrospectively reviewed. Of these patients, we included those fulfilling the CDC criteria for CFS, with a follow-up period greater than one year. We evaluated epidemiological, clinical and evolutive data recorded by their usual physicians. Moreover, the patients were interviewed in order to know their own appreciation with respect to their current clinical status, as well as their present working situation. RESULTS: Twenty-eight patients were included in the present study. Their mean age was 38 +/- 7. Seventy-five percent of them were women. The mean time of clinical follow up was of 3.2 +/- 1.8 years. According to evaluation, 21% of patients improved or became asymptomatic. A similar percentage (28%) of improvement was obtained from the interview. Forty-eight percent of cases had transitory or definitive laboral incapacity. Regarding to prognostic factors, we could not find any statistical differences among the analyzed variables except for marital status. In this variable, married patients had better outcome than unmarried patients. CONCLUSION: CFS is an entity with a poor outcome, since it evolves towards to chronicity in an important number of cases. In addition, strong functional disability may be present, leading frequently to laboral incapacity. PMID- 9395422 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 27-1997. A 38-year-old man with the acquired immunodeficiency syndrome and cavitary pulmonary lesions. PMID- 9395423 TI - [Pancreatic hydatid cyst: review of a case]. PMID- 9395424 TI - Guidelines for the management of nausea and vomiting in pregnancy. PMID- 9395425 TI - [Diagnosis of achalasia using 99m-Tc pertechnetate scintigraphy]. AB - A 73-year-old patient presented a 99mTc scintiscan with a focal tracer accumulation left and caudal of the thyroid gland. Further investigations with sonography, CT, esophagoscopy and barium swallow provided the diagnosis of achalasia as the reason for focal 99mTc retention caudal of the thyroid gland. Explanation for 99mTc accumulation within the esophagus may be the nonspecific accumulation of 99mTc not only in the thyroid gland but also in the salivary glands. Excretion of the tracer follows with the saliva. Structural and motor disorders of the esophagus can thus lead to focal tracer retention within the esophagus simulating thyroid tissue. PMID- 9395426 TI - [Gastric anisakiasis diagnosed by endoscopy]. AB - Anisakiasis is a parasitic infestation of increasing incidence in Spain. The case of a 75-years-old male diagnosed with gastric anisakiasis by endoscopy is presented. This case presents some peculiarities such as the previous ingestion of undercooked microwaved fish and the posterior serologic study (determination of IgE specific for Anisakis simplex) which confirmed the diagnosis. The importance of the endoscopic techniques in the diagnosis and treatment of gastric anisakiasis is emphasized. PMID- 9395427 TI - [Distribution of natural parasites--fungi and microsporidia--in a population of malarial mosquito larva (Diptera: Culicidae) before and after infection by the entomopathogenic bacteria Bacillus thuringiensis israelensis]. AB - The distribution specificity of fungi and microsporidies in natural population of Anopheles messeae Fall. and A. beklemishevi Stegny et Kab. and those which survived after treatment them by Bacillus thuringiensis israelensis (Bti) were observed. Parasitic fungus nonselectively affected individuals of both species and all inversion genotypes of A. messeae. Microsporidia Parathelohania messeae affected males and it has not species and genotypic specificity. The 4-th instar larvae of both species infected by parasitic fungus after treatment them by Bti did not survive. The level of microsporidian infection of A. messeae and A. beklemishevi after Bti treatment was reduced from 1.1 +/- 0.5 to 0.5 +/- 0.3% and from 1.3 +/- 1.3 to 0.7 +/- 0.5% accordingly. A. beklemishevi was registered as Parathelohania messeae host for the first time. The harmonious relationships between malaria mosquitoes and their parasites in natural populations may be destructed by the Bti treatments. PMID- 9395428 TI - Postoperative radiotherapy in high-risk premenopausal women with breast cancer who receive adjuvant chemotherapy. Danish Breast Cancer Cooperative Group 82b Trial. AB - BACKGROUND: Irradiation after mastectomy can reduce locoregional recurrences in women with breast cancer, but whether it prolongs survival remains controversial. We conducted a randomized trial of radiotherapy after mastectomy in high-risk premenopausal women, all of whom also received adjuvant systemic chemotherapy with cyclophosphamide, methotrexate, and fluorouracil (CMF). METHODS: A total of 1708 women who had undergone mastectomy for pathological stage II or III breast cancer were randomly assigned to receive eight cycles of CMF plus irradiation of the chest wall and regional lymph nodes (852 women) or nine cycles of CMF alone (856 women). The median length of follow-up was 114 months. The end points were locoregional recurrence, distant metastases, disease-free survival, and overall survival. RESULTS: The frequency of locoregional recurrence alone or with distant metastases was 9 percent among the women who received radiotherapy plus CMF and 32 percent among those who received CMF alone (P<0.001). The probability of survival free of disease after 10 years was 48 percent among the women assigned to radiotherapy plus CMF and 34 percent among those treated only with CMF (P<0.001). Overall survival at 10 years was 54 percent among those given radiotherapy and CMF and 45 percent among those who received CMF alone (P<0.001). Multivariate analysis demonstrated that irradiation after mastectomy significantly improved disease-free survival and overall survival, irrespective of tumor size, the number of positive nodes, or the histopathological grade. CONCLUSIONS: The addition of postoperative irradiation to mastectomy and adjuvant chemotherapy reduces locoregional recurrences and prolongs survival in high-risk premenopausal women with breast cancer. PMID- 9395429 TI - Nutritional benefits of neonatal screening for cystic fibrosis. Wisconsin Cystic Fibrosis Neonatal Screening Study Group. AB - BACKGROUND: Many patients with cystic fibrosis are malnourished at the time of diagnosis. Whether newborn screening and early treatment may prevent the development of a nutritional deficiency is not known. METHODS: We compared the nutritional status of patients with cystic fibrosis identified by neonatal screening or by standard diagnostic methods. A total of 650,341 newborn infants were screened by measuring immunoreactive trypsinogen on dried blood spots (from April 1985 through June 1991) or by combining the trypsinogen test with DNA analysis (from July 1991 through June 1994). Of 325,171 infants assigned to an early-diagnosis group, cystic fibrosis was diagnosed in 74 infants, including 5 with negative screening tests. Excluding infants with meconium ileus, we evaluated nutritional status for up to 10 years by anthropometric and biochemical methods in 56 of the infants who received an early diagnosis and in 40 of the infants in whom the diagnosis was made by standard methods (the control group). Pancreatic insufficiency was managed with nutritional interventions that included high-calorie diets, pancreatic-enzyme therapy, and fat-soluble vitamin supplements. RESULTS: The diagnosis of cystic fibrosis was confirmed by a positive sweat test at a younger age in the early-diagnosis group than in the control group (mean age, 12 vs. 72 weeks). At the time of diagnosis, the early diagnosis group had significantly higher height and weight percentiles and a higher head-circumference percentile (52nd, vs. 32nd in the control group; P=0.003). The early-diagnosis group also had significantly higher anthropometric indexes during the follow-up period, especially the children with pancreatic insufficiency and those who were homozygous for the deltaF508 mutation. CONCLUSIONS: Neonatal screening provides the opportunity to prevent malnutrition in infants with cystic fibrosis. PMID- 9395430 TI - Bacterial meningitis in the United States in 1995. Active Surveillance Team. AB - BACKGROUND: Before the introduction of the conjugate vaccines, Haemophilus influenzae type b was the major cause of bacterial meningitis in the United States, and meningitis was primarily a disease of infants and young children. We describe the epidemiologic features of bacterial meningitis five years after the H. influenzae type b conjugate vaccines were licensed for routine immunization of infants. METHODS: Data were collected from active, population-based surveillance for culture-confirmed meningitis and other invasive bacterial disease during 1995 in laboratories serving all the acute care hospitals in 22 counties of four states (total population, more than 10 million). The rates were compared with those for 1986 obtained by similar surveillance. RESULTS: On the basis of 248 cases of bacterial meningitis in the surveillance areas, the rates of meningitis (per 100,000) for the major pathogens in 1995 were Streptococcus pneumoniae, 1.1; Neisseria meningitidis, 0.6; group B streptococcus, 0.3; Listeria monocytogenes, 0.2; and H. influenzae, 0.2. Group B streptococcus was the predominant pathogen among newborns, N. meningitidis among children 2 to 18 years old, and S. pneumoniae among adults. Pneumococcal meningitis had the highest case fatality rate (21 percent) and in 36 percent of cases was caused by organisms that were not susceptible to penicillin. From these data, we estimate that 5755 cases of bacterial meningitis were caused by these five pathogens in the United States in 1995, as compared with 12,920 cases in 1986, a reduction of 55 percent. The median age of persons with bacterial meningitis increased greatly, from 15 months in 1986 to 25 years in 1995, largely as a result of a 94 percent reduction in the number of cases of H. influenzae meningitis. CONCLUSIONS: Because of the vaccine related decline in meningitis due to H. influenzae type b, bacterial meningitis in the United States is now a disease predominantly of adults rather than of infants and young children. PMID- 9395431 TI - Surgical treatment for epilepsy: a retrospective Swedish multicenter study. AB - The characteristics of patients suffering from drug resistant epilepsy, including the results of the preoperative evaluation and epilepsy surgery were retrospectively analyzed in a Swedish multicenter 10-year cohort of children and adults. Altogether 152 patients (65 children and 87 adults) treated during the period 1980-1990 in three epilepsy centers were included and followed-up 2 years after surgery. Median age at onset of seizures was 4 years for the children and 12 years for the adults. A localization related epilepsy was present in 85% of the children and in 95% of the adults. The mean number of seizure types in the children was 1.7 (range 1-4) and in the adults 1.8 (range 1-4). The median monthly seizure frequency was 52 and 15 for children and adults respectively. Resective surgery was performed in 143 cases (94 temporal, 31 extratemporal, 9 multilobar and 9 major resection procedures) and palliative procedures in 16 cases (13 callosotomies and 3 stereotactic amygdalotomies). Postoperative neurological deficits were detected in 9% of the patients after temporal lobe resections and in 15% of the patients after extratemporal and multilobar resection procedures. Two years after resective surgery 53% of the children and 49% of the adults were seizure free. Another 25% of the patients had a more than 50% reduction of seizure frequency. In the postoperative non seizure free group of patients there was a negative correlation between decrease in weighted seizure severity and decrease in seizure frequency. This finding stresses the need for including other parameters than seizure frequency when evaluating the outcome of epilepsy surgery. PMID- 9395432 TI - [Dermatologic laser therapy]. PMID- 9395433 TI - DNA methylation directs a time-dependent repression of transcription initiation. AB - BACKGROUND: The regulation of DNA methylation is required for differential expression of imprinted genes during vertebrate development. Earlier studies that monitored the activity of the Herpes simplex virus (HSV) thymidine kinase (tk) gene after injection into rodent cells have suggested that assembly of chromatin influences the methylation-dependent repression of gene activity. Here, we examine the mechanism of methylation-dependent HSV tk gene regulation by direct determination of nucleoprotein organization during the establishment of a transcriptionally silenced state after microinjection of templates with defined methylation states into Xenopus oocyte nuclei. RESULTS: The transcriptional silencing conferred by a methylated DNA segment was not immediate, as methylated templates were initially assembled into active transcription complexes. The eventual loss of DNase I hypersenitive sites and inhibition of transcription at the HSV tk promoter only occurred after several hours. Flanking methylated vector DNA silenced the adjacent unmethylated HSV tk promoter, indicative of a dominant transmissible repression originating from a center of methylation. The resulting repressive nucleoprotein structure silenced transcription in the presence of activators that are able to overcome repression of transcription by nucleosomes. CONCLUSIONS: Silencing of transcription by DNA methylation is achieved at the level of transcription initiation and involves the removal of transcriptional machinery from active templates. This transcriptional repression can occur by indirect mechanisms involving the time-dependent assembly of repressive nucleoprotein complexes, which are able to inhibit transcription more effectively than nucleosomes alone. PMID- 9395434 TI - The lipid transfer activity of phosphatidylinositol transfer protein is sufficient to account for enhanced phospholipase C activity in turkey erythrocyte ghosts. AB - BACKGROUND: The minor membrane phospholipid phosphatidylinositol 4, 5 bisphosphate (PIP2) has been implicated in the control of a number of cellular processes. Efficient synthesis of this lipid from phosphatidylinositol has been proposed to require the presence of a phosphatidylinositol/phosphatidylcholine transfer protein (PITP), which transfers phosphatidylinositol and phosphatidylcholine between membranes, but the mechanism by which PITP exerts its effects is currently unknown. The simplest hypothesis is that PITP replenishes agonist-sensitive pools of inositol lipids by transferring phosphatidylinositol from its site of synthesis to sites of consumption. Recent cellular studies, however, led to the proposal that PITP may play a more active role as a co-factor which stimulates the activity of phosphoinositide kinases and phospholipase C (PLC) by presenting protein-bound lipid substrates to these enzymes. We have exploited turkey erythrocyte membranes as a model system in which it has proved possible to distinguish between the above hypotheses of PITP function. RESULTS: In turkey erythrocyte ghosts, agonist-stimulated PIP2 hydrolysis is initially rapid, but it declines and reaches a plateau when approximately 15% of the phosphatidylinositol has been consumed. PITP did not affect the initial rate of PIP2 hydrolysis, but greatly prolonged the linear phase of PLC activity until at least 70% of phosphatidylinositol was consumed. PITP did not enhance the initial rate of phosphatidylinositol 4-kinase activity but did increase the unstimulated steady-state levels of both phosphatidylinositol 4-phosphate and PIP2 by a catalytic mechanism, because the amount of polyphosphoinositides synthesized greatly exceeded the molar amount of PITP in the assay. Furthermore, when polyphosphoinositide synthesis was allowed to proceed in the presence of exogenous PITP, after washing ghosts to remove PITP before activation of PLC, enhanced inositol phosphate production was observed, whether or not PITP was present in the subsequent PLC assay. CONCLUSION: PITP acts by catalytically transferring phosphatidylinositol down a chemical gradient which is created as a result of the depletion of phosphatidylinositol at its site of use by the concerted actions of the phosphoinositide kinases and PLC. PITP is therefore not a co-factor for the phosphoinositide-metabolizing enzymes present in turkey erythrocyte ghosts. PMID- 9395435 TI - Human p21-activated kinase (Pak1) regulates actin organization in mammalian cells. AB - BACKGROUND: The Rho family GTPases Cdc42, Rac1 and RhoA regulate the reorganization of the actin cytoskeleton induced by extracellular signals such as growth factors. In mammalian cells, Cdc42 regulates the formation of filopodia, whereas Rac regulates lamellipodia formation and membrane ruffling, and RhoA regulates the formation of stress fibers. Recently, the serine/threonine protein kinase p65(pak) autophosphorylates, thereby increasing its catalytic activity towards exogenous substrates. This kinase is therefore a candidate effector for the changes in cell shape induced by growth factors. RESULTS: Here, we report that the microinjection of activated Pak1 protein into quiescent Swiss 3T3 cells induces the rapid formation of polarized filopodia and membrane ruffles. The prolonged overexpression of Pak1 amino-terminal mutants that are unable to bind Cdc42 or Rac1 results in the accumulation of filamentous actin in large, polarized membrane ruffles and the formation of vinculin-containing focal complexes within these structures. This phenotype resembles that seen in motile fibroblasts. The amino-terminal Pak1 mutant displays enhanced binding to the adaptor protein Nck, which contains three Src-homology 3 (SH3) domains. Mutation of a proline residue within a conserved SH3-binding region at the amino terminus of Pak1 interferes with SH3-protein binding and alters the effects of Pak1 on the cytoskeleton. CONCLUSIONS: These results indicate that Pak1, acting through a protein that contains an SH3 domain, regulates the structure of the actin cytoskeleton in mammalian cells, and may serve as an effector for Cdc42 and/or Rac1 in promoting cell motility. PMID- 9395436 TI - Ras signalling is required for inactivation of the tumour suppressor pRb cell cycle control protein. AB - Ras proteins act as molecular switches, responding to signals by entering the active GTP-bound, rather than the inactive GDP-bound, state. The inhibition of normal Ras proteins by microinjection of neutralizing antibody or expression of dominant-negative mutants has shown that Ras signalling is required for growth factors to stimulate DNA synthesis [1] [2], but the link between Ras and the cell cycle machinery is not clear. Regulation of the phosphorylation state of the retinoblastoma protein (pRb), the product of the tumour suppressor gene Rb, is a key event in the progression of cells from G1 phase into S phase. In growth arrested or early G1 cells, pRb is hypophosphorylated and binds to transcription factors of the E2F family [3]. These pRb-E2F complexes act to suppress gene transcription required for entry into DNA synthesis either by preventing E2F from stimulating transcription or by actively repressing transcription [4]. During G1, cyclin-dependent kinases (CDKs) become activated and phosphorylate pRb at multiple sites, leading to the dissolution of pRb-E2F complexes and gene transcription [5]. Here, we have tested the hypothesis that Ras signalling is required for the inactivation of pRb. A neutralizing antibody directed against p21Ras was microinjected into cells derived from mutant mouse embryos that lack Rb or CDK inhibitors (CDKIs). Cells without pRb or the p16 CDKI were more resistant to the inhibitory effects of the anti-Ras antibody. DNA synthesis in some tumour cell lines was completely resistant to the anti-Ras injection, indicating that p21Ras is required for pRb inactivation but also has other functions in cell-cycle progression. PMID- 9395437 TI - Images in clinical medicine. Werner's syndrome. PMID- 9395438 TI - Interprovincial data requirements for local health indicators: the British Columbia experience. AB - Indicators based on the registration of vital events are used to determine the health status of populations. The need for these indicators at the regional and community levels has grown with the trend toward decentralization in the delivery of health services. Such indicators are important because they affect funding and the types of service that are provided. Health status indicators tend to be associated with variables such as the level of urbanization or socioeconomic status. According to four indicators-mortality ratios for all causes of death, mortality ratios for external causes of death, infant mortality ratios, and low birth weight live birth ratios-some areas of British Columbia, specifically along the border with Alberta, have relatively good health, although the characteristics of these regions suggest that this should not be the case. However, a much different picture emerges when vital event data registered in Alberta for residents of these areas of British Columbia are considered. This article shows that for adequate health planning and program implementation, some communities need data from neighbouring provinces. It illustrates the effect of incorporating Alberta data into the development of health status indicators for British Columbia. It also suggests that similar adjustments may be necessary for data compiled in other provinces. PMID- 9395439 TI - The Health Utility Index: measuring health differences in Ontario by socioeconomic status. AB - The positive relationship between socioeconomic status (SES) and longevity has long been established. Comparable evidence exists for SES and morbidity, but observations of this relationship tend to be limited to specific health indicators. In this article, a comprehensive quantitative measure of health status, the Health Utility Index (HUI), is applied to an analysis of the relationship between SES and the health status of people aged 25 and over in Ontario. The HUI, based on a set of questions included in the 1990 Ontario Health Survey (OHS), provides a summary index of the health of each respondent. The OHS data show that lower levels of education, income, and occupation are associated with lower HUI values. Health status differences across SES groups are greater in late middle-age than at younger or older ages, a pattern consistent with the findings of other studies. The development of summary indicators like the HUI is part of a larger effort to construct measures for monitoring the health of Canadians. PMID- 9395440 TI - Causes of death: how the sexes differ. PMID- 9395441 TI - Accidents in Canada, 1988 and 1993. AB - Using data from Statistics Canada's 1988 and 1993 General Social Survey (GSS), this article examines the incidence and consequences of accidents in Canada and the characteristics of respondents aged 15 and over who were involved in them. In 1993, an estimated 3.9 million Canadians reported that they had been involved in 4.8 million accidents in the previous 12 months. Motor vehicle accidents and sports accidents were the most frequent, each accounting for about 27% of incidents, followed by accidents at work (21%) and at home (14%). Accidents were most common among young people, particularly men. However, from 1988 to 1993, there was a decline in the proportion of adults reporting accidents, and the sharpest drop was for the age group most at risk-15- to 24-year-olds. Most of the downturn was attributable to a decrease in the motor vehicle accident rate. Since alcohol is known to be associated with accidents, reduced consumption during the same period may have been partly responsible for the decline in accident rates. Other factors that may have contributed include stricter enforcement of impaired driving legislation and speed limits, and improvements in automobile safety. Nonetheless, despite the decline in accident rates, the toll taken by accidents reported in 1993 was considerable: 80% of accidents caused personal injury, and almost half of these resulted in medical attention in a hospital. Overall, 62% of accidents resulted in activity-loss days, and 29% involved bed-disability days. Hospital utilization costs associated with these accidents in 1993 were about $1.5 billion. As well, about one-third of accidents involved out-of-pocket expenses, totalling $791 million. Moreover, accidents continue to be the leading cause of death among persons under age 44. PMID- 9395442 TI - Mechanism of ion transport across membranes. Bacteriorhodopsin as a prototype for proton pumps. PMID- 9395443 TI - A caveolar complex between the cationic amino acid transporter 1 and endothelial nitric-oxide synthase may explain the "arginine paradox". AB - Immunohistochemistry of porcine pulmonary artery endothelial cells (PAEC) with antibodies specific for caveolin, endothelial nitric-oxide synthase (eNOS), and the arginine transporter (CAT1) demonstrates that all of these proteins co localize in plasma membrane caveolae. When incubated with solubilized PAEC plasma membrane proteins, eNOS-specific antibody immunoprecipitates CAT1-mediated arginine transport. These results document the existence of a caveolar complex between CAT1 and eNOS in PAEC that provides a mechanism for the directed delivery of substrate arginine to eNOS. Direct transfer of extracellular arginine to membrane-bound eNOS accounts for the "arginine paradox" and explains why caveolar localization of eNOS is required for optimal nitric oxide production by endothelial cells. PMID- 9395444 TI - Expression cloning of a novel scavenger receptor from human endothelial cells. AB - Scavenger receptors mediate the endocytosis of chemically modified lipoproteins, such as acetylated low density lipoprotein (Ac-LDL) and oxidized LDL (Ox-LDL), and have been implicated in the pathogenesis of atherosclerosis. The evidence that endothelial cells possess scavenger receptor activity is substantial, and this property is widely used in the isolation of endothelial cells from vascular tissues. In the current study, we have isolated, by expression cloning, the cDNA encoding a novel type of scavenger receptor expressed by endothelial cells (SREC), which mediates the binding and degradation of Ac-LDL. The primary structure of the molecule has no significant homology to other types of scavenger receptors, including the recently cloned endothelial cell Ox-LDL receptor, a member of the C-type lectin family. The cDNA encodes a protein of 830 amino acids with a calculated molecular mass of 85, 735 Da (mature peptide). Chinese hamster ovary cells stably expressing SREC bound 125I-labeled Ac-LDL with high affinity (Kd = 3.0 microg/ml, approximately 1.7 nM) and degraded them via an endocytic pathway. Association of DiII-Ac-LDL were effectively inhibited by Ox-LDL, malondialdehyde-modified LDL, dextran sulfate, and polyinosinic acid, but not by natural LDL and heparin. The cloned receptor has several characteristic domain structures, including an N-terminal extracellular domain with five epidermal growth factor-like cysteine pattern signatures and an unusually long C-terminal cytoplasmic domain (391 amino acids) composed of a Ser/Pro-rich region followed by a Gly-rich region. PMID- 9395445 TI - Sarcospan, the 25-kDa transmembrane component of the dystrophin-glycoprotein complex. AB - The dystrophin-glycoprotein complex is a multisubunit protein complex that spans the sarcolemma and forms a link between the subsarcolemmal cytoskeleton and the extracellular matrix. Primary mutations in the genes encoding the proteins of this complex are associated with several forms of muscular dystrophy. Here we report the cloning and characterization of sarcospan, a unique 25-kDa member of this complex. Topology algorithms predict that sarcospan contains four transmembrane spanning helices with both N- and C-terminal domains located intracellularly. Phylogenetic analysis reveals that sarcospan's arrangement in the membrane as well as its primary sequence are similar to that of the tetraspan superfamily of proteins. Sarcospan co-localizes and co-purifies with the dystrophin-glycoprotein complex, demonstrating that it is an integral component of the complex. We also show that sarcospan expression is dramatically reduced in muscle from patients with Duchenne muscular dystrophy. This suggests that localization of sarcospan to the membrane is dependent on proper dystrophin expression. The gene encoding sarcospan maps to human chromosome 12p11.2, which falls within the genetic locus for congenital fibrosis of the extraocular muscle, an autosomal dominant muscular dystrophy. PMID- 9395446 TI - Correlating Ca2+ responses and secretion in individual RBL-2H3 mucosal mast cells. AB - The role of Ca2+ in stimulus-response coupling in nonexcitable cells is still not well understood. The Ca2+ responses of individual cells are extremely diverse, often displaying marked oscillations, and almost nothing is known about the specific features of these Ca2+ signals that are important for the functional response of a cell. Using the RBL-2H3 mucosal mast cell as a model, we have studied the temporal relationship between changes in intracellular Ca2+ and serotonin secretion at the single-cell level using simultaneous indo-1 photometry and constant potential amperometry. Secretion in response to antigen never occurs until intracellular Ca2+ is elevated, nor is it seen during the first few oscillations in Ca2+. Exocytotic events tend to be clustered around the peaks of oscillations, but excellent secretion is also seen in cells with sustained elevations in Ca2+. Ca2+ release from stores in the absence of influx fails to elicit secretion. If refilling and continued release of Ca2+ from stores is prevented with thapsigargin, Ca2+ influx can still trigger secretion, suggesting that store-associated microdomains of Ca2+ are not required for exocytosis. Our findings demonstrate the importance of an amplitude-encoded Ca2+ signal and Ca2+ influx for stimulus-secretion coupling in these nonexcitable cells. PMID- 9395447 TI - An Eps homology (EH) domain protein that binds to the Ral-GTPase target, RalBP1. AB - Ral proteins constitute a family of small GTPases that can be activated by Ras in cells. In the GTP-bound state, Ral proteins bind to RalBP1, a GTPase-activating protein for CDC42 and Rac GTPases. We have used the two-hybrid system in yeast to clone a cDNA for a novel approximately 85-kDa protein that can bind to an additional site on RalBP1. This newly identified protein contains an Eps homology (EH) domain, which was first detected in the epidermal growth factor (EGF) receptor substrate Eps15. Recently, the EH domain of Eps15 has been shown to bind to proteins containing an asparagine-proline-phenylalanine motif. Moreover, EH domains have been found in proteins involved in endocytosis and/or actin cytoskeleton regulation. The RalBP1 associated Eps-homology domain protein, Reps1, is tyrosine-phosphorylated in response to EGF stimulation of cells. In addition, Reps1 has the capacity to form a complex with the SH3 domains of the adapter proteins Crk and Grb2, which may link Reps1 to an EGF-responsive tyrosine kinase. Thus, Reps1 may coordinate the cellular actions of activated EGF receptors and Ral-GTPases. PMID- 9395448 TI - Characterization of substrate phosphorylation and use of calmodulin mutants to address implications from the enzyme crystal structure of calmodulin-dependent protein kinase I. AB - Calcium/calmodulin (CaM) directly activates CaM-dependent protein kinase I (CaMKI) by binding to the enzyme and indirectly promotes the phosphorylation and synergistic activation of CaMKI by an exogenous kinase. We have evaluated the initial CaM-dependent activation of the unphosphorylated form of CaMKI. The kinetics of bacterially expressed human CaMKI show that the peptide syntide-2 is a relatively poor substrate, whereas the synapsin site-1 peptide is 17-fold more specific. The peptide ADR1G is 400-fold more specific than syntide-2, and its catalytic rate is among the highest reported for a kinase peptide substrate. To understand how CaM activates CaMKI, we have characterized the activation of the enzyme by CaM mutants with substitutions at hydrophobic residues. The point mutant M124Q located in the C-terminal domain of CaM produced a 57-fold increase in the CaM activation constant for CaMKI and suggests the involvement of methionine 124 in an important hydrophobic interaction with tryptophan 303 of CaMKI. Substituting two, three, and five hydrophobic residues in the N-terminal domain of CaM increased the CaM activation constant for CaMKI by 10-190-fold and lowered the maximal enzyme activity by more than 80%. Two of these N-terminal mutants of CaM do not affect the Km for peptide substrate but instead produce a 5 10-fold higher Km for ATP. This result demonstrates the critical role of the N terminal domain of CaM in regulating the access of ATP to CaMKI. PMID- 9395449 TI - COL1A1 transgene expression in stably transfected osteoblastic cells. Relative contributions of first intron, 3'-flanking sequences, and sequences derived from the body of the human COL1A1 minigene. AB - Collagen reporter gene constructs have be used to identify cell-specific sequences needed for transcriptional activation. The elements required for endogenous levels of COL1A1 expression, however, have not been elucidated. The human COL1A1 minigene is expressed at high levels and likely harbors sequence elements required for endogenous levels of activity. Using stably transfected osteoblastic Py1a cells, we studied a series of constructs (pOBColCAT) designed to characterize further the elements required for high level of expression. pOBColCAT, which contains the COL1A1 first intron, was expressed at 50-100-fold higher levels than ColCAT 3.6, which lacks the first intron. This difference is best explained by improved mRNA processing rather than a transcriptional effect. Furthermore, variation in activity observed with the intron deletion constructs is best explained by altered mRNA splicing. Two major regions of the human COL1A1 minigene, the 3'-flanking sequences and the minigene body, were introduced into pOBColCAT to assess both transcriptional enhancing activity and the effect on mRNA stability. Analysis of the minigene body, which includes the first five exons and introns fused with the terminal six introns and exons, revealed an orientation-independent 5-fold increase in CAT activity. In contrast the 3' flanking sequences gave rise to a modest 61% increase in CAT activity. Neither region increased the mRNA half-life of the parent construct, suggesting that CAT specific mRNA instability elements may serve as dominant negative regulators of stability. This study suggests that other sites within the body of the COL1A1 minigene are important for high expression, e.g. during periods of rapid extracellular matrix production. PMID- 9395450 TI - PKC-epsilon is required for mechano-sensitive activation of ERK1/2 in endothelial cells. AB - Mechano-sensitive regulation of endothelial cells (EC) function by shear stress is critical for flow-induced vasodilation and gene expression. Previous studies by our laboratory demonstrated that shear stress activates the 44- and 42-kDa extracellular signal-regulated kinases (ERK1/2) in EC in a time- and force dependent manner. ERK1/2 activation was inhibited by protein kinase C (PKC) down regulation with phorbol 12,13-dibutyrate (1 microM for 24 h) but not by calcium chelation with BAPTA-AM (acetoxymethyl ester of BAPTA) (75 microM for 30 min), suggesting that a novel PKC isoform (delta, epsilon, eta, theta) mediates shear stress-induced ERK1/2 activation. Western blotting with PKC isoform-specific antibodies demonstrated expression of PKC-alpha, -epsilon, and -zeta isoforms in EC. PKC-epsilon was specifically inhibited by transfection with antisense PKC epsilon phosphorothioate oligonucleotides (1,000 nM for 6 h). Antisense treatment decreased PKC-epsilon protein levels by 80 +/- 13% after 72 h and completely inhibited shear stress-stimulated ERK1/2 activation. Scrambled PKC-epsilon oligonucleotides and antisense PKC-alpha and PKC-zeta oligonucleotides had no effect on ERK1/2 activity. PKC-epsilon appeared specific for mechano-sensitive ERK1/2 activation, as antisense PKC-epsilon oligonucleotides did not inhibit ERK1/2 activation by EGF or bradykinin but did inhibit ERK1/2 activation upon EC adhesion to fibronectin. These results define a pathway for shear stress-mediated ERK1/2 activation and establish a new function for PKC-epsilon as part of a mechano-sensitive signal transduction pathway in EC. PMID- 9395451 TI - Molecular and biochemical characterization of an endo-beta-1,3- glucanase of the hyperthermophilic archaeon Pyrococcus furiosus. AB - We report here the first molecular characterization of an endo-beta-1,3-glucanase from an archaeon. Pyrococcus furiosus is a hyperthermophilic archaeon that is capable of saccharolytic growth. The isolated lamA gene encodes an extracellular enzyme that shares homology with both endo-beta-1,3- and endo-beta-1,3-1,4 glucanases of the glycosyl hydrolase family 16. After deletion of the N-terminal leader sequence, a lamA fragment encoding an active endo-beta-1,3-glucanase was overexpressed in Escherichia coli using the T7-expression system. The purified P. furiosus endoglucanase has highest hydrolytic activity on the beta-1,3-glucose polymer laminarin and has some hydrolytic activity on the beta-1,3-1,4 glucose polymers lichenan and barley beta-glucan. The enzyme is the most thermostable endo-beta-1,3-glucanase described up to now; it has optimal activity at 100-105 degrees C. In the predicted active site of glycosyl hydrolases of family 16 that show predominantly endo-beta-1,3-glucanase activity, an additional methionine residue is present. Deletion of this methionine did not change the substrate specificity of the endoglucanase, but it did cause a severe reduction in its catalytic activity, suggesting a structural role of this residue in constituting the active site. High performance liquid chromatography analysis showed in vitro hydrolysis of laminarin by the endo-beta-1,3-glucanase proceeds more efficiently in combination with an exo-beta-glycosidase from P. furiosus (CelB). This most probably reflects the physiological role of these enzymes: cooperation during growth of P. furiosus on beta-glucans. PMID- 9395452 TI - Purification and characterization of recombinant catalase-peroxidase, which confers isoniazid sensitivity in Mycobacterium tuberculosis. AB - The Mycobacterium tuberculosis katG gene encodes a dual-function enzyme called catalase-peroxidase, which confers sensitivity in M. tuberculosis to isonicotinic acid hydrazide. We have constructed a system for the high level expression of a recombinant form of this enzyme by amplifying the katG gene from the pYZ56 construct (1) and subcloning into a vector suitable for expression in Escherichia coli. The resulting plasmid, pTBCP, produced the catalase-peroxidase in large quantities, corresponding to 30% of total cell protein. The enzyme has been purified to homogeneity and appears to be a dimer in the native form. Using either hydrogen peroxide or t-butyl hydroperoxide and 2,2'-azino-bis(3 ethylbenzthiazoline-6-sulfonic acid) as substrates, kcat and Km values have been obtained for both catalatic and peroxidatic activities, respectively. The availability of significant quantities of an active, folded, recombinant form of M. tuberculosis catalase-peroxidase should thus facilitate future studies of its role in drug activation and antibiotic resistance. PMID- 9395453 TI - Muscarinic receptor-mediated dual regulation of ADP-ribosyl cyclase in NG108-15 neuronal cell membranes. AB - Cyclic ADP-ribose (cADP-ribose) is an endogenous modulator of ryanodine-sensitive Ca2+ release channels. An unsolved question is whether or not cADP-ribose mediates intracellular signals from hormone or neurotransmitter receptors. The first step in this study was to develop a TLC method to measure ADP-ribosyl cyclase, by which conversion of [3H]NAD+ to [3H]cADP-ribose was confirmed in COS 7 cells overexpressing human CD38. A membrane fraction of NG108-15 neuroblastoma x glioma hybrid cells possessed ADP-ribosyl cyclase activity measured by TLC. Carbamylcholine increased this activity by 2.6-fold in NG108-15 cells overexpressing m1 or m3 muscarinic acetylcholine receptors (mAChRs), but inhibited it by 30-52% in cells expressing m2 and/or m4 mAChRs. Both of these effects were mimicked by GTP. Pretreatment of cells with cholera toxin blocked the activation, whereas pertussis toxin blocked the inhibition. Application of carbamylcholine caused significant decreases in NAD+ concentrations in untreated m1-transformed NG108-15 cells, but an increase in cholera toxin-treated cells. These results suggest that mAChRs couple to ADP-ribosyl cyclase within cell membranes via trimeric G proteins and can thereby control cellular function by regulating cADP-ribose formation. PMID- 9395454 TI - Differential regulation of the transcriptional activity of the orphan nuclear receptor NGFI-B by membrane depolarization and nerve growth factor. AB - The immediate-early gene NGFI-B (also called nur77) encodes an orphan nuclear receptor that activates transcription through a unique response element (NBRE). NGFI-B is rapidly induced and modified via phosphorylation by a variety of stimuli that induce cells to differentiate or to proliferate. We have shown that the in vitro phosphorylation of Ser350 located within the "A-box," a motif necessary for DNA binding by NGFI-B, results in a decrease in the binding of NGFI B to its response element (Hirata, Y., Kiuchi, K., Chen, H.-C., Milbrandt, J., and Guroff, G. (1993) J. Biol. Chem. 268, 24808-24812). We show here that nerve growth factor (NGF)-induced changes in the in vivo phosphorylation of Ser350 accompany transcriptional deactivation of NGFI-B in PC12 cells, that membrane depolarization and NGF treatment cause differential phosphorylation of NGFI-B, and that the transcriptional activation caused by exogenous expression of NGFI-B or membrane depolarization can be inhibited by NGF treatment. In addition, the mutation of Ser350 to Ala abolished the inhibitory effect of NGF on the transcriptional activation of NGFI-B in PC12 cells. These data could provide new insights into the regulation of transcriptional activity required for some neurons to switch from activity-dependent survival to neurotrophin-dependent survival during development. PMID- 9395455 TI - Heparan sulfate proteoglycans participate in hepatic lipaseand apolipoprotein E mediated binding and uptake of plasma lipoproteins, including high density lipoproteins. AB - High density lipoprotein (HDL) particles and HDL cholesteryl esters are taken up by both receptor-mediated and non-receptor-mediated pathways. Here we show that cell surface heparan sulfate proteoglycans (HSPG) participate in hepatic lipase (HL)- and apolipoprotein (apo) E-mediated binding and uptake of mouse and human HDL by cultured hepatocytes. The HL secreted by HL-transfected McA-RH7777 cells enhanced both HDL binding at 4 degrees C (approximately 2-4-fold) and HDL uptake at 37 degrees C (approximately 2-5-fold). The enhanced binding and uptake of HDL were partially inhibited by the 39-kDa protein, an inhibitor of low density lipoprotein receptor-related protein (LRP), but were almost totally blocked by heparinase, which removes the sulfated glycosaminoglycan chains from HSPG. Therefore, HL may mediate the uptake of HDL by two pathways: an HSPG-dependent LRP pathway and an HSPG-dependent but LRP-independent pathway. The HL-mediated binding and uptake of HDL were only minimally reduced when catalytically inactive HL or LRP binding-defective HL was substituted for wild-type HL, indicating that much of the HDL uptake required neither HL binding to the LRP nor lipolytic processing. To study the role of HL in facilitating the selective uptake of cholesteryl esters, we used HDL into which radiolabeled cholesteryl ether had been incorporated. HL increased the selective uptake of HDL cholesteryl ether; this enhanced uptake was reduced by more than 80% by heparinase but was unaffected by the 39-kDa protein. Like HL, apoE enhanced the binding and uptake of HDL (approximately 2-fold) but had little effect on the selective uptake of HDL cholesteryl ether. In the presence of HL, apoE did not further increase the uptake of HDL, and at a high concentration apoE impaired or decreased the HL mediated uptake of HDL. Therefore, HL and apoE may utilize similar (but not identical) binding sites to mediate HDL uptake. Although the relative importance of cell surface HSPG in the overall metabolism of HDL in vivo remains to be determined, cultured hepatocytes clearly displayed an HSPG-dependent pathway that mediates the binding and uptake of HDL. This study also demonstrates the importance of HL in enhancing the binding and uptake of remnant and low density lipoproteins via an HSPG-dependent pathway. PMID- 9395456 TI - Bovine proenteropeptidase is activated by trypsin, and the specificity of enteropeptidase depends on the heavy chain. AB - Enteropeptidase, also known as enterokinase, initiates the activation of pancreatic hydrolases by cleaving and activating trypsinogen. Enteropeptidase is synthesized as a single-chain protein, whereas purified enteropeptidase contains a approximately 47-kDa serine protease domain (light chain) and a disulfide linked approximately 120-kDa heavy chain. The heavy chain contains an amino terminal membrane-spanning segment and several repeated structural motifs of unknown function. To study the role of heavy chain motifs in substrate recognition, secreted variants of recombinant bovine proenteropeptidase were constructed by replacing the transmembrane domain with a signal peptide. Secreted variants containing both the heavy chain (minus the transmembrane domain) and the catalytic light chain (pro-HL-BEK (where BEK is bovine enteropeptidase)) or only the catalytic domain (pro-L-BEK) were expressed in baby hamster kidney cells and purified. Single-chain pro-HL-BEK and pro-L-BEK were zymogens with extremely low catalytic activity, and both were activated readily by trypsin cleavage. Trypsinogen was activated efficiently by purified enteropeptidase from bovine intestine (Km = 5.6 microM and kcat = 4.0 s-1) and by HL-BEK (Km = 5.6 microM and kcat = 2.2 s-1), but not by L-BEK (Km = 133 microM and kcat = 0.1 s-1); HL-BEK cleaved trypsinogen at pH 5.6 with 520-fold greater catalytic efficiency than did L-BEK. Qualitatively similar results were obtained at pH 8.4. In contrast to this striking difference in trypsinogen recognition, the small synthetic substrate Gly Asp-Asp-Asp-Asp-Lys-beta-naphthylamide was cleaved with similar kinetic parameters by both HL-BEK (Km = 0.27 mM and kcat = 0.07 s-1) and L-BEK (Km = 0.60 mM and kcat = 0.06 s-1). The presence of the heavy chain also influenced the rate of reaction with protease inhibitors. Bovine pancreatic trypsin inhibitor preferred HL-BEK (initial Ki = 99 nM and final Ki* = 1.8 nM) over L-BEK (Ki = 698 nM and Ki* = 6.2 nM). Soybean trypsin inhibitor exhibited a reciprocal pattern, inhibiting L-BEK (Ki* = 1.6 nM), but not HL-BEK. These kinetic data indicate that the enteropeptidase heavy chain has little influence on the recognition of small peptides, but strongly influences macromolecular substrate recognition and inhibitor specificity. PMID- 9395457 TI - A novel white laccase from Pleurotus ostreatus. AB - Two laccase isoenzymes (POXA1 and POXA2) produced by Pleurotus ostreatus were purified and fully characterized. POXA1 and POXA2 are monomeric glycoproteins with 3 and 9% carbohydrate content, molecular masses of about 61 and 67 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis, of about 54 and 59 kDa by gel filtration in native conditions, and of 61 kDa by matrix-assisted laser desorption ionization mass spectrometry (only for POXA1) and pI values of 6.7 and 4.0, respectively. The N terminus and three tryptic peptides of POXA1 have been sequenced, revealing clear homology with laccases from other microorganisms, whereas POXA2 showed a blocked N terminus. The stability of POXA2 as a function of temperature was particularly low, whereas POXA1 showed remarkable high stability with respect to both pH and temperature. Both enzymes oxidize syringaldazine and ABTS (2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) together with a variety of different substituted phenols and aromatic amines with the concomitant reduction of oxygen, but POXA1 is unable to oxidize guaiacol. Both enzymes were strongly inhibited by sodium azide and thioglycolic acid but not by EDTA. UV/visible absorption spectra, atomic adsorption, and polarographic data indicated the presence of 4 copper atoms/mol of POXA2 but only one copper, two zinc, and one iron atoms were found/mol of POXA1. The neutral pI and the anomalous metal content of POXA1 laccase render this enzyme unique in its structural characteristics. The lack of typical absorbance at 600 nm allows its classification as a "white" laccase. PMID- 9395458 TI - Effects of intravesicular H+ and extracellular H+ and Zn2+ on insulin secretion in pancreatic beta cells. AB - The effects of extracellular Zn2+ and pH and intravesicular pH on insulin and 5 hydroxytryptamine (5-HT) secretion from pancreatic beta cells were investigated. Insulin and 5-HT secretion from single cells was detected by amperometry as a series of current spikes corresponding to detection of multimolecular packets secreted by exocytosis. Spike width was used as a measure of the kinetics of clearance from the cell and the area of spikes as a measure of amount released. Changes in extracellular pH from 6.9 to 7.9 caused insulin spikes to become narrower with no change in area, whereas the same treatments had no effect on 5 HT secretion. Treatment of cells with Bafilomycin A1 or N-ethylmaleimide, both of which are expected to increase intravesicular pH by inhibiting V-type H+-ATPase, had no effect on 5-HT secretion but caused insulin spikes to become more narrow. These results indicate that exposure to high pH, whether intravesicular or extracellular, accelerates release of insulin during exocytosis without affecting the amount of insulin released. Increasing extracellular Zn2+ concentration from 0 to 25 microM increased the width and decreased the area of insulin spikes without affecting 5-HT secretion. Zn2+ effects were likely exerted through a common-ion effect on Zn2+-insulin dissociation. It was concluded that intravesicular storage conditions and extracellular ions can affect free insulin concentration in the vicinity of beta cells during secretion. PMID- 9395459 TI - Identification and cloning of the membrane-associated serine protease, hepsin, from mouse preimplantation embryos. AB - Previous studies have suggested the existence of a membrane-associated serine protease expressed by mammalian preimplantation embryos. In this study, we have identified hepsin, a type II transmembrane serine protease, in early mouse blastocysts. Mouse hepsin was highly homologous to the previously identified human and rat cDNAs. Two isoforms, differing in their cytoplasmic domains, were detected. The tissue distribution of mouse hepsin was similar to that seen in humans, with prominent expression in liver and kidney. In mouse embryos, hepsin expression was observed in the two-cell stage, reached a maximal level at the early blastocyst stage, and decreased subsequent to blastocyst hatching. Expression of a soluble form of hepsin revealed its ability to autoactivate in a concentration-dependent manner. Catalytically inactive soluble hepsin was unable to autoactivate. These results suggest that hepsin may be the first serine protease expressed during mammalian development, making its ability to autoactivate critical to its function. PMID- 9395460 TI - Activation of PKCalpha downstream from caspases during apoptosis induced by 7 hydroxystaurosporine or the topoisomerase inhibitors, camptothecin and etoposide, in human myeloid leukemia HL60 cells. AB - We previously demonstrated that the anticancer agent and protein kinase C (PKC) inhibitor 7-hydroxystaurosporine (UCN-01) induces apoptosis independently of p53 and protein synthesis in HL60 cells. We now report the associated changes of PKC isoforms. PKCalpha, betaI, betaII, delta, and zeta activities were measured after immunoprecipitation of cytosols from UCN-01-treated HL60 cells. UCN-01 had no effect on PKCzeta and inhibited kinase activity of PKCbetaI, betaII, and delta. PKCalpha activity was initially inhibited at 1 h, and subsequently increased as cells underwent apoptosis 3 h after the beginning of UCN-01 treatment. Camptothecin (CPT) and etoposide (VP-16) also markedly enhanced PKCalpha activity during apoptosis in HL60 cells. However, CPT did not affect PKCbetaI, betaII and zeta, and activated PKCdelta. PKCalpha activation was not due to increased protein levels or proteolytic cleavage but was associated with PKCalpha autophosphorylation in vitro and increased phosphorylation in vivo. We also found that not only PKC delta but also PKC betaI was proteolytically activated in HL60 cells during apoptosis. The PKCalpha activation and hyperphosphorylation were abrogated by N-benzyloxycarbonyl-Val-Ala-Asp(O-methyl)-fluoromethylketone (z-VAD fmk) under conditions that abrogated apoptosis. z-VAD-fmk also prevented PKCdelta and betaI proteolytic activation. Together these findings suggest that caspases regulate PKC activity during apoptosis in HL60 cells. At least two modes of activation were observed: hyperphosphorylation for PKCalpha and proteolytic activation for PKC delta and betaI. PMID- 9395461 TI - Exchange of subunit interfaces between recombinant adult and fetal hemoglobins. Evidence for a functional inter-relationship among regions of the tetramer. AB - The inter-relationship between the interior subunit interfaces and the exterior diphosphoglycerate (DPG) binding region of the hemoglobin tetramer and the effects of a specific N-terminal acetylation on tetramer assembly have been evaluated. Tetrameric fetal hemoglobin F in the liganded state was found to dissociate to dimers much less than previously appreciated, i.e. about 70 times less than adult hemoglobin A (Kd = 0.01 microM and 0.68 microM, for HbF and HbA, at pH 7.5, respectively) over the pH range 6.2-7.5, whereas HbF1, in which the N termini of the gamma-chains are acetylated, dissociates like HbA. To determine whether this feature of HbF could be transferred to hemoglobin A, the single amino acid difference in their alpha1beta2/alpha1gamma2 interfaces and the 4 amino acid differences in their alpha1beta1/alpha1gamma1 interfaces have been substituted in HbA to those in HbF. This pentasubstituted recombinant HbA/F had the correct molecular weight as determined by mass spectrometry, the expected mobility on isoelectric focusing, the calculated amino acid composition, and normal circular dichroism properties, oxygen binding, and cooperativity. Although HbA/F has the same amino acid side chains that bind DPG as HbA, its diminished response to 2,3-DPG resembled that of HbF. However, its tetramer-dimer dissociation constant (Kd = 0.14 microM) was between that of HbA and HbF despite the fact that it was composed entirely of HbF subunit interfaces. The results indicate that regions of the tetramer distant from the tetramer-dimer interface influence its dissociation and, reciprocally, that the interfaces affect regions involved in the binding of allosteric regulators, suggesting flexible long range inter-relationships in hemoglobin. PMID- 9395462 TI - The effect of apolipoprotein A-II on the structure and function of apolipoprotein A-I in a homogeneous reconstituted high density lipoprotein particle. AB - In this study we examined the effects of apoA-II on the structure and function of apoA-I in homogeneous reconstituted HDL (rHDL). First, we measured the binding of apoA-II to apoA-I-rHDL, containing dipalmitoylphosphatidylcholine or palmitoyloleoylphosphatidylcholine, and the degree of apoA-I displacement at various ratios of apolipoproteins. Using fluorescence methods, we determined that apoA-II binding is rapid, irreversible, and associated with apoA-I displacement only when the molar ratio of apoA-II/apoA-I is greater than 1:2. Next, we used the stable apoA-II/apoA-I-rHDL complex at the apoA-II/apoA-I ratio of 1:2 to examine its physical properties, apoA-I structure, and reactivity with lecithin:cholesterol acyltransferase (LCAT). Using chemical cross-linking in conjunction with fluorescence and electrophoretic methods, we demonstrated that the conformation of apoA-I must be flexible to allow apoA-II binding to the apoA I-rHDL particles and showed that the hybrid particles have an unchanged Stokes diameter. Fluorescence and circular dichroism measurements revealed little or no change in the secondary structure or in the N-terminal domain of apoA-I, but showed a marked destabilization of apoA-I to denaturation by guanidine hydrochloride. Limited tryptic digestion indicated that the central region of apoA-I becomes accessible to proteolysis in the hybrid particles. Together, these results suggest that amphipathic alpha-helices of apoA-II replace four central helices of one apoA-I molecule (residues approximately 99-187) in the complex and in the process destabilize apoA-I. Thus, apoA-II binding at physiologic ratios may not completely displace apoA-I from HDL, but may provide a reservoir of easily exchangeable apoA-I. Finally, we showed that the reaction of the hybrid HDL with LCAT was inhibited 2-5-fold, relative to apoA-I-rHDL, due to a corresponding increase in the apparent Km value. This suggests that LCAT binding to the hybrid particles is sterically hindered by the excess protein (portions of apoA-I and apoA-II not bound to lipid). Therefore, apoA-II can modulate the reaction of HDL with LCAT by decreasing LCAT binding to hybrid particles and making the enzyme available for reaction with other substrates. PMID- 9395463 TI - Purification and specificity of beta1,2-xylosyltransferase, an enzyme that contributes to the allergenicity of some plant proteins. AB - The enzyme that transfers D-xylose from UDP-xylose to the beta-linked mannose of plant N-linked oligosaccharides was purified about 51,000-fold to apparent homogeneity from soybean microsomes. On SDS gels, two proteins of 56 and 59 kDa were detected and both were labeled to the same extent by the photoaffinity label, 5-N3-UDP-[32P]xylose. Labeling of both proteins was inhibited by cold UDP xylose, but not by UDP-glucose. The amount of 5-N3-UDP-[32P]xylose that bound to the two protein bands was greatly increased in the presence of oligosaccharide acceptors. The best acceptor for xylose transfer and for stimulation of UDP xylose binding was GlcNAc2Man3GlcNAc2-T, but GlcNAc1Man3GlcNAc2, with the GlcNAc on the 3-branch, was also a good acceptor and a good stimulator. A number of other N-linked oligosaccharides were poor acceptors, especially those with galactose units at the nonreducing termini. Many of the properties of this enzyme have been described, and the product of the reaction of UDP-xylose and GlcNAc2Man3(GlcNAc)2 was characterized as GlcNAcbeta1, 2Manalpha1, 6(GlcNAcbeta1,2Manalpha1,3)(Xylbeta1,2)Manbeta1, 4GlcNA c2-T by chemical and NMR methods. PMID- 9395464 TI - Inhibitory effects of specific apolipoprotein C-III isoforms on the binding of triglyceride-rich lipoproteins to the lipolysis-stimulated receptor. AB - ApoC-III overexpression in mice results in severe hypertriglyceridemia due primarily to a delay in the clearance of triglyceride-rich lipoproteins. We have, in primary cultures of rat hepatocytes, characterized a lipolysis-stimulated receptor (LSR). The apparent number of LSR that are available on rat liver plasma membranes is negatively correlated with plasma triglyceride concentrations measured in the fed state. We therefore proposed that the primary physiological role of the LSR is to contribute to the cellular uptake of triglyceride-rich lipoproteins. We have now tested the effect of apoC-III on the binding of triglyceride-rich lipoproteins to LSR. Supplementation of 125I-very low density lipoprotein (VLDL) with apoC-III inhibited the LSR-mediated binding, internalization, and degradation of 125I-VLDL in primary cultures of rat hepatocytes. Studies using isolated rat liver plasma membranes showed that enrichment of human VLDL and chylomicrons with synthetic or purified human apoC III decreased their binding to the LSR by about 40%. Supplementation of triglyceride-rich lipoproteins under the same conditions with human apoC-II had no such inhibitory effect, despite the fact that this apoprotein bound as efficiently as apoC-III to these particles. Preincubation of LDL with apoC-III did not modify its binding to LSR. Partitioning studies using 125I-apoC-III showed that this lack of effect was due to apoC-III's inability to efficiently associate with LDL. Purified human apoC-III1 was as efficient as the synthetic nonsialylated form of apoC-III in inhibiting binding of VLDL to LSR. However, despite a 2-fold greater binding of apoC-III2 to VLDL, this isoform was a less efficient inhibitor of the binding of VLDL to LSR than apoC-III1 or nonsialylated apoC-III. Desialylation of apoC-III2 by treatment with neuraminidase increased the inhibition of VLDL binding to LSR to a level similar to that observed with apoC-III1 and nonsialylated apoC-III. We propose that apoC-III regulates in part the rate of removal of triglyceride-rich particles by inhibiting their binding to the LSR, and that the level of inhibition is determined by the degree of apoC-III sialylation. PMID- 9395465 TI - The carboxyl terminus of the human calcium receptor. Requirements for cell surface expression and signal transduction. AB - The G-protein-coupled calcium receptor plays a key role in extracellular calcium homeostasis. To examine the role of the membrane-spanning domains and the approximately 200-residue cytoplasmic carboxyl terminus of the calcium receptor in cell-surface expression and signal transduction, we transfected HEK-293 cells with a series of truncation and carboxyl-terminal missense mutants and analyzed expression by immunoblotting, glycosidase digestion, intact cell immunoassay, and extracellular calcium-stimulated phosphoinositide hydrolysis assay. Two truncation mutants terminating at residues 706 and 802 within the second and third intracellular loops, respectively, were not properly glycosylated, failed to reach the cell-surface, and showed no calcium response, indicating that mutant receptors with the full extracellular domain but only three or five transmembrane domains are improperly folded and/or processed. Truncation mutants terminating at residues 888 and 903 within the carboxyl terminus were equivalent to the wild type in all assays, whereas mutants truncated at residues 865 and 874 showed no response to calcium, despite only approximately 25% reduction in cell-surface expression. Mutants with a full-length carboxyl terminus but with residues between positions 874 and 888 replaced with alanines showed either no (Ala875, Ala876, and Ala879) or significantly reduced (Ala881-Ala883) calcium response at levels of cell-surface expression equivalent to those of the wild-type receptor. These results indicate that deletion of the majority of the carboxyl terminus is compatible with normal processing, cell-surface expression, and signal transduction of the receptor. The truncation and alanine substitution mutants identify a small region between residues 874 and 888 critical for normal signal transduction by the receptor. PMID- 9395466 TI - Identification and quantitation of the fatty acids composing the CoA ester pool of bovine retina, heart, and liver. AB - Several proteins found in retinal photoreceptor cells (guanylate cyclase activating protein, protein kinase A, recoverin, and transducin) are N-terminally modified with the fatty acids 12:0, 14:0, 14:1n-9, and 14:2n-6, whereas similar proteins in other tissues contain only 14:0. It has been hypothesized that the acyl-CoA pool of the retina contains amounts of 12:0, 14:1n-9, and 14:2n-6 elevated over 14:0, in comparison to other tissues, and this accounts for the specificity of N-terminal fatty acylation. To test this hypothesis, we performed fatty acid analysis on total acyl-CoAs purified from bovine retina (light adapted), heart, and liver. We also examined the N- and S-linked fatty acid composition of the total protein pools from these tissues. Acyl-CoAs were prepared from heart, liver, and retina and separated by high performance liquid chromatography (HPLC). Identities of peaks were based on HPLC of standard 12:0, 14:0, 14:1n-9, and 14:2n-6 CoAs. Total protein was subjected to base hydrolysis followed by acidic methanolysis to release S- and N-linked fatty acids, respectively, and fatty acid phenacyl esters were prepared for HPLC analysis. Retina had levels of 12:0 (2.7 +/- 2.1%), 14:1n-9 (2.9 +/- 2.2%), and 14:2n-6 (1.6 +/- 0.7%) CoAs below that of 14:0 CoA (7.0 +/- 1.8%). Likewise, heart levels of 14:2n-6 CoA (3.7 +/- 0.1%) were near and 12:0 (2.6 +/- 0. 6%) and 14:1n-9 (0.7 +/- 0.3%) CoAs were below that of 14:0 CoA (3.8 +/- 1.0%). Liver had levels of 12:0 (16.1 +/- 5.7%) and 14:2n-6 (8.1 +/- 1.2%) CoAs above and 14:1n-9 CoA (1.2 +/- 0.6%) below that of 14:0 CoA (5.9 +/- 0.8%). Fatty acid analysis of total protein showed that all tissues contained S-linked 16:0, 18:0, and 18:1n-9. Retina proteins contained N-linked 14:0, 14:1n-9, and 14:2n-6, whereas heart and liver had only 14:0. Our findings do not support the hypothesis that the CoA ester pool of the retina is enriched with 12:0, 14:1n-9, and 14:2n-6 over 14:0, in comparison to other tissues. This suggests that alternative models must be considered for the regulation of N-terminal fatty acylation of proteins in photoreceptor cells. PMID- 9395467 TI - cea5, a structurally divergent member of the murine carcinoembryonic antigen gene family, is exclusively expressed during early placental development in trophoblast giant cells. AB - The carcinoembryonic antigen (CEA) gene family encodes a large family of glycoproteins. Some are probably involved in the homeostasis/development of epithelial cells and granulocyte activation, while others e.g. the pregnancy specific glycoproteins, are expressed in the placenta and are essential for a positive outcome of pregnancy. In this paper, we have characterized cea5, a member of the murine CEA gene family. RNase protection and in situ hybridization analyses revealed that Cea5 mRNA is exclusively synthesized in primary and secondary trophoblast giant cells of the placenta only during early stages of development. Full-length Cea5 cDNA was obtained by a reverse transcription polymerase chain reaction using day 10.5 post-coitum placental RNA. The 1.6 kilobase pair (kb) Cea5 mRNA encodes a secreted glycoprotein with a predicted size of 30 kDa. It is composed of a leader peptide (L), one immunoglobulin (Ig) variable or N, and one Ig constant-like or A domain. This domain organization is unique within the human and murine CEA families. Two overlapping cosmid clones covering 54 kb of the cea5 gene locus were mapped. cea5 consists of three exons (L, N, A/3'-untranslated region exon) located within a 4-kb region. rnCGM2, the rat cea5 counterpart, exhibits the same restricted expression pattern. This together with their exceptional conservation within the rat and murine CEA families and their absence from the human CEA family suggests that cea5 and rnCGM2 are of functional importance for rodent placental development. PMID- 9395468 TI - Developmental regulation of the sulfation profile of chondroitin sulfate chains in the chicken embryo brain. AB - Developmentally regulated and cell type-specific expression of distinct sulfated glycosaminoglycan structures on cell surface proteoglycans is increasingly recognized as providing information relevant to cell-cell interactions and differentiation in developing organisms. In this report, developmental regulation of both the sulfation profile of chondroitin sulfate chains and activities of chondroitin 4-sulfotransferase (C4ST) and chondroitin 6-sulfotransferase (C6ST) were evaluated in embryonic chicken brain. The results revealed that the sulfation profile and the sulfotransferase activities changed markedly with development, and these alterations were precisely coordinated. Specifically, the proportions of both chondroitin 6-sulfate to 4-sulfate and C6ST to C4ST activities progressively decreased with development. In addition, the total amounts of both chondroitin sulfate chains and the sulfotransferase activities were highest during early embryonic stages and decreased sharply as the development reached completion. The developmental expression of the C6ST gene was also found to parallel the developmental down-regulation of both the C6ST activity and the chondroitin 6-sulfate structure. These findings suggest that the developmentally regulated expression of the sulfotransferases is a predominant factor for stage-specific regulation of chondroitin sulfate structures. PMID- 9395469 TI - The carboxyl terminus of the Saccharomyces cerevisiae succinate dehydrogenase membrane subunit, SDH4p, is necessary for ubiquinone reduction and enzyme stability. AB - The succinate dehydrogenase (SDH) of Saccharomyces cerevisiae is composed of four nonidentical subunits encoded by the nuclear genes SDH1, SDH2, SDH3, and SDH4. The hydrophilic subunits, SDH1p and SDH2p, comprise the catalytic domain involved in succinate oxidation. They are anchored to the inner mitochondrial membrane by two small, hydrophobic subunits, SDH3p and SDH4p, which are required for electron transfer and ubiquinone reduction. Comparison of the deduced primary sequence of the yeast SDH4p subunit to SDH4p subunits from other species reveals the presence of an unusual 25-30 amino acid carboxyl-terminal extension following the last predicted transmembrane domain. The extension is predicted to be on the cytoplasmic side of the inner mitochondrial membrane. To investigate the extension's function, three truncations were created and characterized. The results reveal that the carboxyl-terminal extension is necessary for respiration and growth on nonfermentable carbon sources, for ubiquinone reduction, and for enzyme stability. Combined with inhibitor studies using a ubiquinone analog, our results suggest that the extension and more specifically, residues 128-135 are involved in the formation of a ubiquinone binding site. Our findings support a two-ubiquinone binding site model for the S. cerevisiae SDH. PMID- 9395470 TI - The chicken genome contains two functional nonallelic beta1,4 galactosyltransferase genes. Chromosomal assignment to syntenic regions tracks fate of the two gene lineages in the human genome. AB - Two distinct but related groups of cDNA clones, CKbeta4GT-I and CKbeta4GT-II, have been isolated by screening a chicken hepatoma cDNA library with a bovine beta1,4-galactosyltransferase (beta4GT) cDNA clone. CKbeta4GT-I is predicted to encode a type II transmembrane glycoprotein of 41 kDa with one consensus site for N-linked glycosylation. CKbeta4GT-II is predicted to encode a type II transmembrane glycoprotein of 43 kDa with five potential N-linked glycosylation sites. At the amino acid level, the coding regions of CKbeta4GT-I and CKbeta4GT II are 52% identical to each other and 62 and 49% identical, respectively, to bovine beta4GT. Despite this divergence in amino acid sequence, high levels of expression of each cDNA in Trichoplusia ni insect cells demonstrate that both CKbeta4GT-I and CKbeta4GT-II encode an alpha-lactalbumin-responsive, UDP galactose:N-acetylglucosamine beta4-galactosyltransferase. An analysis of CKbeta4GT-I and CKbeta4GT-II genomic clones established that the intron positions within the coding region are conserved when compared with each other, and these positions are identical to the mouse and human beta4GT genes. Thus CKbeta4GT-I and CKbeta4GT-II are the result of the duplication of an ancestral gene and subsequent divergence. CKbeta4GT-I maps to chicken chromosome Z in a region of conserved synteny with the centromeric region of mouse chromosome 4 and human chromosome 9p, where beta4-galactosyltransferase (EC 2.4.1.38) had previously been mapped. Consequently, during the evolution of mammals, it is the CKbeta4GT-I gene lineage that has been recruited for the biosynthesis of lactose. CKbeta4GT II maps to a region of chicken chromosome 8 that exhibits conserved synteny with human chromosome 1p. An inspection of the current human gene map of expressed sequence tags reveals that there is a gene noted to be highly similar to beta4GT located in this syntenic region on human chromosome 1p. Because both the CKbeta4GT-I and CKbeta4GT-II gene lineages are detectable in mammals, duplication of the ancestral beta4-galactosyltransferase gene occurred over 250 million years ago in an ancestral species common to both mammals and birds. PMID- 9395471 TI - Modulation of insulin receptor substrate-1 tyrosine phosphorylation and function by mitogen-activated protein kinase. AB - Increased serine phosphorylation of insulin receptor substrate-1 (IRS-1) has been observed in several systems to correlate with a decreased ability of the insulin receptor to tyrosine-phosphorylate this endogenous substrate and to inhibit its subsequent association with phosphatidylinositol 3-kinase. In the present studies we have examined the potential role of the mitogen-activated protein (MAP) kinase in the increased serine phosphorylation of IRS-1 observed in human embryonic kidney cells treated with an activator of protein kinase C, phorbol 12-myristate 13-acetate. First, recombinantly produced kinase was shown to phosphorylate intact IRS-1 in a way that decreased the ability of isolated insulin receptor to phosphorylate the tyrosines recognized by the SH2 domains of the phosphatidylinositol 3-kinase. Second, an inhibitor of MAP kinase activation, PD98059, blocked the phorbol 12-myristate 13-acetate-induced inhibition of the insulin-stimulated increase in IRS-1 associated phosphatidylinositol 3-kinase. Third, activation of MAP kinase in intact cells via a regulatable upstream kinase, a RAF:estrogen receptor construct, could also inhibit the insulin stimulated increase in IRS-1-associated phosphatidylinositol 3-kinase. Fourth, an in gel kinase assay showed that MAP kinase was the primary renaturable kinase in cell extracts capable of phosphorylating an IRS-1 fusion protein. Finally, IRS-1 was found to associate in coprecipitation studies with endogenous MAP kinase. These studies implicate MAP kinase as one of the kinases capable of phosphorylating and regulating IRS-1 tyrosine phosphorylation. PMID- 9395472 TI - Thermodynamic analysis of the interaction between the 0.5beta Fv fragment and the RP135 peptide antigen derived from the V3 loop of HIV-1 gp120. AB - The Fv fragment of the 0.5beta monoclonal antibody has recently been constructed, expressed, and purified. It binds with nanomolar affinity to the immunogenic RP135 peptide that is derived from the principal neutralizing determinant of HIV 1 in the third hypervariable region of gp120. Here, we analyzed the temperature dependence of binding of the 0.5beta Fv fragment to the RP135 peptide and a series of mutants thereof. Our results show that there is almost complete enthalpy-entropy compensation in the effects of mutations in the peptide on binding to the Fv, indicating that the mutations do not change the binding mechanism. There is good correlation, for residues within the antigenic epitope, between mutational effects on DeltaCp and calculated values of DeltaDeltaCp based on the extent of burial of polar and non-polar surface areas of amino acids. The value of DeltaCp for the binding of the 0.5beta Fv fragment to the wild-type RP135 peptide is found to be -5.0 (+/- 0.9) kcal K-1 mol-1 in the presence of 0.1% Tween-20 but only -0.1 (+/- 0.9) kcal K-1 mol-1 in its absence. This result has important implications for the successful application of the structural parameterization approach to predicting changes in heat capacity that accompany binding reactions carried out in the presence of detergent or protein-stabilizing agents. PMID- 9395473 TI - Site-directed disulfide mapping of helices M4 and M6 in the Ca2+ binding domain of SERCA1a, the Ca2+ ATPase of fast twitch skeletal muscle sarcoplasmic reticulum. AB - In an attempt to define the spatial relationships among SERCA1a transmembrane helices M4, M5, M6, and M8, involved in Ca2+ binding, all six cysteine residues were removed from predicted transmembrane sequences by substitution with Ser or Ala. The cysteine-depleted protein retained 44% of wild type Ca2+ transport activity. Pairs of cysteine residues were then reintroduced to determine whether their juxtaposition would result in the formation of disulfide cross-links between transmembrane helices. In initial studies designed to map the juxtaposition of Ca2+ binding residues, Cys was substituted for Glu309 or Gly310 in transmembrane sequence M4, in combination with the substitution of Cys for Glu771 in M5; for Asn796, Thr799, or Asp800 in M6; or for Glu908 in M8. These double mutants all retained the capacity to form a phosphoenzyme intermediate from Pi (but not from ATP in the presence of Ca2+), and in all but mutants E309C/N796C and G310C/N796C, phosphoenzyme formation was insensitive to 100 microM Ca2+. These results support the view that both Glu309 and Asn796 contribute to Ca2+ binding site II, which is not required for conversion of E2, the substrate for Pi phosphorylation, to E1. Cross-linking in mutants E309C/N796C and G310C/D800C established reference points for the orientation of M4 and M6 relative to each other and provided the basis for the prediction of potential additional cross-links. Strong links were formed with the pairs T317C/A804C and T317C/L807C near the cytoplasmic ends of the two helices and with A305C/L792C and A305C/L793C near the lumenal ends. These combined results support the conclusion that M4 and M6 form a right-handed coiled-coil structure that forms part of the pathway of Ca2+ translocation. In addition to providing a possible explanation for the mutation sensitivity of several pairs of residues in these helices, the proposed association of M4 and M6 supports a new model for the orientation of the two Ca2+ binding sites among transmembrane helices M4, M5, and M6. PMID- 9395474 TI - The molecular chaperone function of the secretory vesicle cysteine string proteins. AB - The "J" domains of eukaryotic DnaJ-like proteins specify interaction with various Hsp70s. The conserved tripeptide, HPD, present in all J domains has been shown to be important for the interaction between yeast and bacterial DnaJ/Hsp70 protein pairs. We have characterized mutations in the HPD motif of the synaptic vesicle protein cysteine-string protein (Csp). Mutation of the histidine (H43Q) or aspartic acid (D45A) residues of this motif reduced the ability of Csp to stimulate the ATPase activity of mammalian Hsc70. The H43Q and D45A mutant proteins were not able to stimulate the ATPase activity of Hsc70 to any significant extent. The mutant proteins were characterized by competition assays, tryptic digestion analysis, and direct binding analysis from which it was seen that these proteins were defective in binding to Hsc70. Thus, the HPD motif of Csp is required for binding to Hsc70. We also analyzed the interaction between Csp and a model substrate protein, denatured firefly luciferase. Both Csp1 and the C-terminally truncated isoform Csp2 were able to prevent aggregation of heat denatured luciferase, and they also cooperated with Hsc70 to prevent aggregation. In addition, complexes of Csp1 or Csp2 with Hsc70 and luciferase were isolated, confirming that these proteins interact and that Csps can bind directly to denatured proteins. Csp1 and Csp2 isoforms must differ in some aspect other than interaction with Hsc70 and substrate protein. These results show that both Csp1 and Csp2 can bind a partially unfolded protein and act as chaperones. This suggests that Csps may have a general chaperone function in regulated exocytosis. PMID- 9395475 TI - Two NFAT transcription factor binding sites participate in the regulation of CD95 (Fas) ligand expression in activated human T cells. AB - Antigen receptor engagement on T lymphocytes activates transcription factors important for stimulating cytokine gene expression. This is critical for clonal expansion of antigen-specific T cells and propagation of immune responses. Additionally, under some conditions antigen receptor stimulation initiates apoptosis of T lymphocytes through the induced expression of CD95 ligand and its receptor. Here we demonstrate that the transcription factor, NFAT, which is critical for the inducible expression of many cytokine genes, also plays a critical role in the regulation of T cell receptor-mediated CD95 ligand expression. Two sites within the CD95 ligand promoter, identified through DNase I footprinting, bind NFAT proteins from nuclear extracts of activated T cells. Although both sites appear important for optimal expression of CD95 ligand in activated T cells, mutational analysis suggests that the distal NFAT site plays a more significant role. Furthermore, these sites do not appear to be required for constitutive CD95 ligand expression in Sertoli cells. PMID- 9395476 TI - Acyl-coenzyme A causes Ca2+ release in pancreatic acinar cells. AB - The regulation of cytosolic Ca2+ is important for a variety of cell functions. One non-inositol 1,4,5-trisphosphate (IP3) compound that may regulate Ca2+ is palmitoyl-coenzyme A (CoA), a fatty acid-CoA that is reported to cause Ca2+ release from intracellular stores of oocytes, myocytes, and hepatocytes. To study the role of palmitoyl-CoA in the pancreatic acinar cell, rat pancreatic acini were isolated by collagenase digestion, permeablized with streptolysin O, and the release of Ca2+ from internal stores was measured with fura-2. Palmitoyl-CoA released Ca2+ from internal stores (EC50 = 14 microM). The palmitoyl-CoA sensitive pool was distinct from, and overlapping with the IP3-sensitive Ca2+ pool. The effects of submaximal doses of IP3 or cyclic ADP-ribose plus palmitoyl CoA were additive. Fatty acid-CoA derivatives with carbon chain lengths of 16-18 were the most potent and efficacious. Ryanodine and caffeine or elevated resting [Ca2+] sensitized the Ca2+ pool to the actions of palmitoyl-CoA. Fatty acid-CoA levels in pancreatic acini were measured by extraction with 2 propanol/acetonitrile, followed by separation and quantification using reverse phase high performance liquid chromatography, and were found to be 10.17 +/- 0.93 nmol/mg protein. These data suggest the presence of an IP3-insensitive palmitoyl CoA-sensitive Ca2+ store in pancreatic acinar cells and suggest that palmitoyl CoA may be needed for Ca2+-induced Ca2+ release. PMID- 9395477 TI - Positional preferences of ionizable residues in Gly-X-Y triplets of the collagen triple-helix. AB - Collagens contain a high amount of charged residues involved in triple-helix stability, fibril formation, and ligand binding. The contribution of charged residues to stability was analyzed utilizing a host-guest peptide system with a single Gly-X-Y triplet embedded within Ac(Gly-Pro-Hyp)3-Gly-X-Y-(Gly-Pro-Hyp)4 Gly-Gly-NH2. The ionizable residues Arg, Lys, Glu, and Asp were incorporated into the X position of Gly-X-Hyp; in the Y position of Gly-Pro-Y; or as pairs of oppositely charged residues occupying X and Y positions. The Gly-X-Hyp peptides had similar thermal stabilities, only marginally less stable than Gly-Pro-Hyp, whereas Gly-Pro-Y peptides showed a wide thermal stability range (Tm = 30-45 degrees C). The stability of peptides with oppositely charged residues in the X and Y positions appears to reflect simple additivity of the individual residues, except when X is occupied by a basic residue and Y = Asp. The side chains of Glu, Lys, and Arg have the potential to form hydrogen bonds with available peptide backbone carbonyl groups within the triple-helix, whereas the shorter Asp side chain does not. This may relate to the unique involvement of Asp residues in energetically favorable ion pair formation. These studies clarify the dependence of triple-helix stability on the identity, position, and ionization state of charged residues. PMID- 9395478 TI - Fibronectin type III repeats mediate RGD-independent adhesion and signaling through activated beta1 integrins. AB - Many cell-surface and extracellular matrix proteins contain multiple modular domains known as fibronectin type III (FNIII) repeats. Cells adhere to the extracellular matrix proteins fibronectin and tenascin in part by the interaction of certain integrins with the Arg-Gly-Asp (RGD) sequence, displayed on specific FNIII repeats. We have found that, after experimental activation of beta1 integrins, a number of cell types adhere and spread on FNIII repeats lacking RGD, derived from extracellular matrix proteins and cytokine receptors. Interaction between individual FNIII domains and beta1 integrins mediates focal adhesion kinase phosphorylation and subsequent stress fiber and focal contact formation. These data suggest that many FNIII-containing proteins may bind and signal through activated beta1 integrins, dramatically expanding the potential for integrin-dependent intercellular and cell-matrix communication. PMID- 9395479 TI - Structural analysis of the human BIN1 gene. Evidence for tissue-specific transcriptional regulation and alternate RNA splicing. AB - BIN1 is a putative tumor suppressor that was identified through its interaction with the MYC oncoprotein. To begin to identify elements of BIN1 whose alteration may contribute to malignancy, we cloned and characterized the human BIN1 gene and promoter. Nineteen exons were identified in a region of >54 kilobases, six of which were alternately spliced in a cell type-specific manner. One alternately spliced exon encodes part of the MYC-binding domain, suggesting that splicing controls the MYC-binding capacity of BIN1 polypeptides. Four other alternately spliced exons encode amphiphysin-related sequences that were included in brain specific BIN1 species, also termed amphiphysin isoforms or amphiphysin II. The 5' flanking region of BIN1 is GC-rich and lacks a TATA box but directs transcriptional initiation from a single site. A approximately 0. 9-kilobase fragment from this region was sufficient for basal transcription and transactivation by MyoD, which may account for the high levels of BIN1 observed in skeletal muscle. This study lays the foundation for genetic and epigenetic investigations into the role of BIN1 in normal and neoplastic cell regulation. PMID- 9395480 TI - Mints, Munc18-interacting proteins in synaptic vesicle exocytosis. AB - Munc18-1 is a neuronal protein that interacts with syntaxin 1 and is required for synaptic vesicle exocytosis. We have now identified two Munc18-1-interacting proteins called Mint1 and Mint2 that may mediate the function of Munc18-1. Mint proteins are detectable only in brain and are composed of an N-terminal sequence that binds Munc18-1, a middle phosphotyrosine-binding domain, and two C-terminal PDZ domains thought to attach proteins to the plasma membrane. In brain, Mint proteins are part of a multimeric complex containing Munc18-1 and syntaxin that likely functions as an intermediate in synaptic vesicle docking/fusion. The phosphotyrosine-binding domain specifically binds to phosphatidylinositol phosphates known to be produced during vesicle exocytosis (Hay, J. C., Fisette, P. L., Jenkins, G. H., Fukami, K., Takonawa, T., Anderson, R. A., and Martin, T. F. J. (1995) Nature 374, 173-177). Our data suggest a model whereby local production of phosphatidylinositol phosphates may trigger the binding of vesicles to the active zone via the Mint.Munc18-1 complex in conjunction with syntaxin 1. PMID- 9395481 TI - Chicken ovalbumin upstream promoter-transcription factor interacts with estrogen receptor, binds to estrogen response elements and half-sites, and inhibits estrogen-induced gene expression. AB - Chicken ovalbumin upstream promoter-transcription factor (COUP-TF) was identified as a low abundance protein in bovine uterus that co-purified with estrogen receptor (ER) in a ligand-independent manner and was separated from the ER by its lower retention on estrogen response element (ERE)-Sepharose. In gel mobility shift assays, COUP-TF bound as an apparent dimer to ERE and ERE half-sites. COUP TF bound to an ERE half-site with high affinity, Kd = 1.24 nM. In contrast, ER did not bind a single ERE half-site. None of the class II nuclear receptors analyzed, i.e. retinoic acid receptor, retinoid X receptor, thyroid receptor, peroxisome proliferator-activated receptor, or vitamin D receptor, were constituents of the COUP-TF.DNA binding complex detected in gel mobility shift assays. Direct interaction of COUP-TF with ER was indicated by GST "pull-down" and co-immunoprecipitation assays. The nature of the ER ligand influenced COUP-TF ERE half-site binding. When ER was liganded by the antiestrogen 4 hydroxytamoxifen (4-OHT), COUP-TF-half-site interaction decreased. Conversely, COUP-TF transcribed and translated in vitro enhanced the ERE binding of purified estradiol (E2)-liganded ER but not 4-OHT-liganded ER. Co-transfection of ER expressing MCF-7 human breast cancer cells with an expression vector for COUP-TFI resulted in a dose-dependent inhibition of E2-induced expression of a luciferase reporter gene under the control of three tandem copies of EREc38. The ability of COUP-TF to bind specifically to EREs and half-sites, to interact with ER, and to inhibit E2-induced gene expression suggests COUP-TF regulates ER action by both direct DNA binding competition and through protein-protein interactions. PMID- 9395482 TI - Phosphoenolpyruvate carboxykinase (GTP) gene transcription and hyperglycemia are regulated by glucocorticoids in genetically obese db/db transgenic mice. AB - The molecular mechanisms underlying increased hepatic phosphoenolpyruvate carboxykinase (PEPCK) gene transcription and gluconeogenesis in type II diabetes are largely unknown. To examine the involvement of glucocorticoids and the cis acting insulin response sequence (IRS, -416/-407) in the genetically obese db/db mouse model, we generated crosses between C57BL/KsJ-db/+ mice and transgenic mice that express -460 or -2000 base pairs of the rat PEPCK gene promoter containing an intact or mutated IRS, linked to a reporter gene. Transgenic mice expressing the intact PEPCK(460)-CRP (C-reactive protein) transgene bred to near homozygosity at the db locus were obese, hyperinsulinemic, and developed fasting hyperglycemia (389 +/- 26 mg/100 ml) between 4 and 10 weeks of age. Levels of CRP reporter gene expression were increased 2-fold despite severe hyperinsulinemia compared with non-diabetic non-obese transgenic mice. Reporter gene expression was also increased 2-fold in transgenic obese diabetic db/db mice bearing a mutation in the IRS, -2000(IRS)-hGx, compared with non-obese non-diabetic transgenic 2000(IRS)-hGx mice. Treatment of obese diabetic db/db transgenic mice with the glucocorticoid receptor blocker RU 486 decreased plasma glucose by 50% and reduced PEPCK, GLUT2, glucose-6-phosphatase, tyrosine aminotransferase, CRP, and hGx reporter gene expression to levels similar to those of non-obese normoglycemic transgenic mice. Taken together, these results establish that -460 bp of 5'-flanking sequence is sufficient to mediate the induction of PEPCK gene transcription in genetically obese db/db mice during the development of hyperglycemia. The results further demonstrate that the mechanism underlying increased expression of gluconeogenic enzymes in the db/db mouse requires the action of glucocorticoids and occurs independently of factors acting through the PEPCK IRS (-416/-407) promoter binding site. PMID- 9395483 TI - Heterodimerization-independent functions of cell death regulatory proteins Bax and Bcl-2 in yeast and mammalian cells. AB - The pro-apoptotic protein Bax can homodimerize with itself and heterodimerize with the anti-apoptotic protein Bcl-2, but the significance of these protein protein interactions remains unclear. Alanine substitution mutations were created in a well conserved IGDE motif found within the BH3 domain of Bax (residues 66 69) and the resulting mutant Bax proteins were tested for ability to homodimerize with themselves and to heterodimerize with Bcl-2. Correlations were made with cell death induction by these mutants of Bax both in mammalian cells where Bax may function through several mechanisms, and in yeast where Bax may exert its lethal actions through a more limited repertoire of mechanisms perhaps related to its ability to form ion channels in intracellular membranes. Two of the mutants, Bax(D68A) and Bax(E69A), retained the ability to homodimerize but failed to interact with Bcl-2 as determined by yeast two-hybrid assays and co immunoprecipitation analysis using transfected mammalian cells. The Bax(E69A) protein exhibited a lethal phenotype in yeast, which could be specifically suppressed by co-expression of Bcl-2, despite its failure to dimerize with Bcl-2. Both the Bax(D68A) and Bax(E69A) proteins induced apoptosis when overexpressed in human 293 cells, despite an inability to bind to Bcl-2. Moreover, co-expression of Bcl-2 with Bax(D68A) and Bax(E69A) rescued mammalian cells from apoptosis. In contrast, a mutant of Bax lacking the IGDE motif, Bax(DeltaIGDE), was incapable of either homodimerizing with itself or heterodimerizing with Bcl-2 and was inactive at promoting cell death in either yeast or mammalian cells. Although failing to interact with Bcl-2, the Bax(D68A) and Bax(E69A) mutants retained the ability to bind to Bid, a putative Bax-activating member of the Bcl-2 family, and collaborated with Bid in inducing apoptosis. When taken together with previous observations, these findings indicate that (i) Bax can induce apoptosis in mammalian cells irrespective of heterodimerization with Bcl-2 and (ii) Bcl-2 can rescue both mammalian cells and yeast from the lethal effects of Bax without heterodimerizing with it. However, these results do not exclude the possibility that BH3-dependent homodimerization of Bax or interactions with Bax activators such as Bid may either assist or be required for the cell death-inducing mechanism of this protein. PMID- 9395484 TI - Immunochemical visualization and quantitation of cyclic AMP in single cells. AB - Adenosine 3':5'-cyclic monophosphate (cAMP) is a key second messenger in signaling pathways governing many cellular processes. To define the subcellular localization and relative abundance of cAMP, we developed a novel immunochemical approach based on acrolein fixation to visualize cAMP within cells. We describe here the fixation and immobilization of cAMP within cells and the production of specific, high titer polyclonal antibodies that recognize cAMP. Relative levels of cAMP immunofluorescence were quantitated in glial cells (oligodendrocytes, astrocytes, Schwann cells, and glioma cells) that were either untreated or treated with activators of endogenous adenylyl cyclase to raise cAMP levels. In treated cells, cAMP immunofluorescence is strongly localized in the perinuclear cytoplasm. PMID- 9395485 TI - Cloning and targeted deletion of the mouse fetuin gene. AB - We proposed that the alpha2-Heremans Schmid glycoprotein/fetuin family of serum proteins inhibits unwanted mineralization. To test this hypothesis in animals, we cloned the mouse fetuin gene and generated mice lacking fetuin. The gene consists of seven exons and six introns. The cystatin-like domains D1 and D2 of mouse fetuin are encoded by three exons each, whereas a single terminal exon encodes the carboxyl-terminal domain D3. The promoter structure is well conserved between rat and mouse fetuin genes within the regions shown to bind transcription factors in the rat system. Expression studies demonstrated that mice homozygous for the gene deletion lacked fetuin protein and that mice heterozygous for the null mutation produced roughly half the amount of fetuin protein produced by wild-type mice. Fetuin-deficient mice were fertile and showed no gross anatomical abnormalities. However, the serum inhibition of apatite formation was compromised in these mice as well as in heterozygotes. In addition, some homozygous fetuin deficient female ex-breeders developed ectopic microcalcifications in soft tissues. These results corroborate a role for fetuin in serum calcium homeostasis. The fact that generalized ectopic calcification did not occur in fetuin-deficient mice proves that additional inhibitors of phase separation exist in serum. PMID- 9395486 TI - Matrix metalloproteinase-8 is expressed in rheumatoid synovial fibroblasts and endothelial cells. Regulation by tumor necrosis factor-alpha and doxycycline. AB - Neutrophil collagenase (matrix metalloproteinase-8 or MMP-8) is regarded as being synthesized exclusively by polymorphonuclear neutrophils (PMN). However, in vivo MMP-8 expression was observed in mononuclear fibroblast-like cells in the rheumatoid synovial membrane. In addition, we detected MMP-8 mRNA expression in cultured rheumatoid synovial fibroblasts and human endothelial cells. Up regulation of MMP-8 was observed after treatment of the cells with either tumor necrosis factor-alpha (10 ng/ml) or phorbol 12-myristate 13-acetate (10 nM). Western analysis showed a similar regulation at the protein level. The size of secreted MMP-8 was 50 kDa, which is about 30 kDa smaller than MMP-8 from PMN. Conditioned media from rheumatoid synovial fibroblasts contained both type I and II collagen degrading activity. However, degradation of type II collagen, but not that of type I collagen, was completely inhibited by 50 microM doxycycline, suggesting specific MMP-8 activity. In addition, doxycycline down-regulated MMP-8 induction, at both the mRNA and protein levels. Thus MMP-8 exerts markedly wider expression in human cells than had been thought previously, implying that PMN are not the only source of cartilage degrading activity at arthritic sites. The inhibition of both MMP-8 activity and synthesis by doxycycline provides an incentive for further studies on the clinical effects of doxycycline in the treatment of rheumatoid arthritis. PMID- 9395487 TI - Disruption of the gene encoding the mitogen-regulated translational modulator PHAS-I in mice. AB - PHAS-I is the prototype of a group of eIF4E-binding proteins that can regulate mRNA translation in response to hormones and growth factors. To investigate the importance of PHAS-I in the physiology of the intact animal, we disrupted the PHAS-I gene in mice. Tissues and cells derived from the knockout mice contained no detectable PHAS-I protein. A related protein, PHAS-II, and eIF4E were readily detectable in tissues from these animals, but neither appeared to be changed in a compensatory manner. Mice lacking PHAS-I appeared normal at birth. However, male knockout mice weighed approximately 10% less than controls at all ages, whereas female weights were similar to those of controls. Both males and females were fertile. Tissues from adult animals appeared to be normal by routine histological staining techniques, as were routine blood cell counts and chemistries. Fibroblasts derived from PHAS-I-deficient mouse embryos exhibited normal rates of growth and overall protein synthesis, responded normally to serum stimulation of ornithine decarboxylase activity and cell growth, and rapamycin inhibition of cell growth. Under these experimental conditions, PHAS-I is apparently not required for the normal development and reproductive behavior of female mice, but is required for normal body weight in male mice; the mechanisms responsible for this phenotype remain to be determined. PMID- 9395488 TI - Role of translocation in the activation and function of protein kinase B. AB - We have investigated the role of subcellular localization in the regulation of protein kinase B (PKB) activation. The myristoylation/palmitylation motif from the Lck tyrosine kinase was attached to the N terminus of protein kinase B to alter its subcellular location. Myristoylated/palmitylated (m/p)-PKBalpha was associated with the plasma membrane of transfected cells, whereas the wild-type kinase was mostly cytosolic. The activity of m/p-PKBalpha was 60-fold higher compared with the unstimulated wild-type enzyme, and could not be stimulated further by growth factors or phosphatase inhibitors. In vivo 32P labeling and mutagenesis demonstrated that m/p-PKBalpha activity was due to phosphorylation on Thr308 and Ser473, that are normally induced on PKB following stimulation of the cells with insulin or insulin-like growth factor-1 (IGF-1). A dominant negative form of phosphoinositide 3-kinase (PI3-K) did not affect m/p-PKBalpha activity. The pleckstrin homology (PH) domain of m/p-PKBalpha was not required for its activation or phosphorylation on Thr308 and Ser473, suggesting that this domain may serve as a membrane-targeting module. Consistent with this view, PKBalpha was translocated to the plasma membrane within minutes after stimulation with IGF-1. This translocation required the PH domain and was sensitive to wortmannin. Our results indicate that PI3-K activity is required for translocation of PKB to the plasma membrane, where its activation occurs through phosphorylation of the same sites that are induced by insulin or IGF-1. Following activation the kinase detached from the membrane and translocated to the nucleus. PMID- 9395489 TI - Self-assembly of laminin isoforms. AB - The alpha, beta, and gamma subunits of basement membrane laminins can combine into different heterotrimeric molecules with either three full short arms (e.g. laminins-1-4), or molecules containing one (laminins-6-9) or more (laminin-5) short arm truncations. Laminin-1 (alpha1beta1gamma1), self-assembles through a calcium-dependent thermal gelation requiring binding interactions between N terminal short arm domains, forming a meshwork polymer thought to contribute to basement membrane architecture (Yurchenco, P. D., and Cheng, Y. S. (1993) J. Biol. Chem. 268, 17286-17299). However, it has been unclear whether other isoforms share this property, and if so, which ones. To begin to address this, we evaluated laminin-2 (alpha2beta1gamma1), laminin-4 (alpha2beta2gamma1), laminin-5 (alpha3Abeta3gamma2), and laminin-6 (alpha3Abeta1gamma1). The first two isoforms were found to self-aggregate in a concentration- and temperature-dependent manner and a close self-assembly relationship among laminins-1, -2, and -4 were demonstrated by: (a) polymerization of all three proteins was inhibited by EDTA and laminin-1 short arm fragments, (b) polymerization of laminin-1 was inhibited by fragments of laminins-2 and -4, (c) laminin-2 and, to a lesser degree, laminin 4, even well below their own critical concentration, co-aggregated with laminin 1, evidence for co-polymerization. Laminin-5, on the other hand, neither polymerized nor co-polymerized with laminin-1. Laminin-6 failed to co-aggregate with laminin-1 at all concentrations evaluated, evidence for a lack of a related self-assembly activity. The data support the hypothesis that all three short arms are required for self-assembly and suggest that the short arm domain structure of laminin isoforms affect their architecture-forming properties in basement membranes. PMID- 9395490 TI - A new mechanism-based radical intermediate in a mutant R1 protein affecting the catalytically essential Glu441 in Escherichia coli ribonucleotide reductase. AB - The invariant active site residue Glu441 in protein R1 of ribonucleotide reductase from Escherichia coli has been engineered to alanine, aspartic acid, and glutamic acid. Each mutant protein was structurally and enzymatically characterized. Glu441 contributes to substrate binding, and a carboxylate side chain at position 441 is essential for catalysis. The most intriguing results are the suicidal mechanism-based reaction intermediates observed when R1 E441Q is incubated with protein R2 and natural substrates (CDP and GDP). In a consecutive reaction sequence, we observe at least three clearly discernible steps: (i) a rapid decay (k1 >/= 1.2 s-1) of the catalytically essential tyrosyl radical of protein R2 concomitant with formation of an early transient radical intermediate species, (ii) a slower decay (k2 = 0.03 s-1) of the early intermediate concomitant with formation of another intermediate with a triplet EPR signal, and (iii) decay (k3 = 0.004 s-1) of the latter concomitant with formation of a characteristic substrate degradation product. The characteristics of the triplet EPR signal are compatible with a substrate radical intermediate (most likely localized at the 3'-position of the ribose moiety of the substrate nucleotide) postulated to occur in the wild type reaction mechanism as well. PMID- 9395491 TI - Specific recognition of an rU2-N15-rU motif by VP55, the vaccinia virus poly(A) polymerase catalytic subunit. AB - VP55, the vaccinia poly(A) polymerase catalytic subunit, interacts with oligonucleotide primers via two uridylate recognition sites (Deng, L., and Gershon, P. D. (1997) EMBO J. 16, 1103-1113). Here, we show that the cognate RNA sequence comprises a 5'-rU2-N15-rU-3' motif (where N = any deoxyribo or ribonucleotide), embedded within oligonucleotide primers 29-30 nucleotides (nt), or greater, in length. Nine residues separate the 3'-most ribouridylate of the optimally positioned motif from the primer 3'-OH. A ribose sugar at the extreme 3'-terminal nucleotide of the primer is absolutely required for VP55's adenylyltransferase activity, but not for stable VP55-RNA interaction. A ribose at position -3 markedly stimulates both adenylyltransferase activity and stable binding. The use of uridine analogs indicated (i) those functional groups of the uracil base which contribute to stable VP55-primer interaction, and (ii) that VP55's ability to discriminate uracil from cytosine stems largely from the requirement for a protonated N3 nitrogen within the pyrimidine ring. The rU2-N15 rU motif was identified within the uridylate-rich 3' end of a naturally occurring vaccinia mRNA. However, oligonucleotides whose only internal uridylates comprised the motif supported only a 3-5-nt processive burst of oligo(A) tail addition, as opposed to the approximately 30-35-nt burst observed with the naturally occurring 3' end. PMID- 9395492 TI - Probing the G-protein regulation of GIRK1 and GIRK4, the two subunits of the KACh channel, using functional homomeric mutants. AB - In heart, G-protein-activated channels are complexes of two homologous proteins, GIRK1 and GIRK4. Expression of either protein alone results in barely active or non-active channels, making it difficult to assess the individual contribution of each subunit to the channel complex. The residue Phe137, located within the H5 region of GIRK1, is critical to the synergy between GIRK1 and GIRK4 (Chan, K. W., Sui, J. L., Vivaudou, M., and Logothetis, D. E. (1996) Proc. Natl. Acad. Sci. U. S. A. 93, 14193-14198). By modifying this residue or the matching residue of GIRK4, Ser143, we have been able to generate mutant proteins that produced large inwardly rectifying, G-protein-modulated currents when expressed alone in Xenopus oocytes. The enhanced activity of the heterologous expression of each of two active mutants, GIRK1(F137S) and GIRK4(S143T), was not caused by association with an endogenous oocyte channel subunit, and these mutants did not display apparent differences in the ability to localize to the cell surface compared with their wild-type counterparts. When these functional mutant channels were compared individually with wild-type heteromeric channels, they responded with only small differences to a number of maneuvers involving coexpression with muscarinic receptors, G-protein betagamma subunits, wild-type or mutated G-protein alpha subunits, and active protomers of pertussis toxin. These experiments, which confirmed the crucial, though not exclusive, role of Gbetagamma in regulating channel activity, demonstrated that GIRK1(F137S) and GIRK4(S143T), and by extrapolation their wild-type counterparts, interact in a qualitatively similar way with G-protein subunits. These findings suggest that functionally important sites of interaction with G-proteins are likely to be located within the homologous regions of GIRK1 and GIRK4 rather than within the divergent terminal regions. They also raise the question of the functional advantage of a heteromeric over homomeric design for G-protein-gated channels. PMID- 9395493 TI - beta-dystrobrevin, a new member of the dystrophin family. Identification, cloning, and protein associations. AB - Dystrophin, the protein disrupted in Duchenne muscular dystrophy, is one of several related proteins that are key components of the submembrane cytoskeleton. Three dystrophin-related proteins (utrophin, dystrophin-related protein-2 (DRP2), and dystrobrevin) have been described. Here, we identify a human gene on chromosome 2p22-23 that encodes a novel protein, beta-dystrobrevin, with significant homology to the other known dystrobrevin (now termed alpha dystrobrevin). Sequence alignments including this second dystrobrevin strongly support the concept that two distinct subfamilies exist within the dystrophin family, one composed of dystrophin, utrophin, and DRP2 and the other composed of alpha- and beta-dystrobrevin. The possibility that members of each subfamily form distinct protein complexes was examined by immunopurifying dystrobrevins and dystrophin. A beta-dystrobrevin antibody recognized a protein of the predicted size (71 kDa) that copurified with the dystrophin short form, Dp71. Thus, like alpha-dystrobrevin, beta-dystrobrevin is likely to associate directly with dystrophin. alpha- and beta-dystrobrevins failed to copurify with each other, however. These results suggest that members of the dystrobrevin subfamily form heterotypic associations with dystrophin and raise the possibility that pairing of a particular dystrobrevin with dystrophin may be regulated, thereby providing a mechanism for assembly of distinct submembrane protein complexes. PMID- 9395494 TI - Role of asymmetric phosphate neutralization in DNA bending by PU.1. AB - The PU.1 transcription factor is a member of the Ets family of DNA binding proteins. PU.1 binds to DNA via a loop-helix-loop domain and functions in the differentiation of hematopoietic cells. The structure of a PU.1-DNA complex was recently reported (Kodandapani, R., Pio, F., Ni, C.-Z., Piccialli, G., Klemsz, M., McKercher, S., Maki, R., and Ely, K. (1996) Nature 380, 456-460). The DNA in this complex is deformed by 8 degrees as it curves around the protein. The pattern of electrostatic contacts between PU.1, and its DNA binding site suggests that laterally asymmetric phosphate neutralization accompanies PU.1 binding. Because of our previous studies showing that such neutralization can induce bending in naked DNA, we have explored the effect of phosphate neutralization by substituting neutral methylphosphonate internucleoside linkages at relevant positions within DNA containing the PU.1 binding sequence. Consistent with the prediction that DNA will collapse toward its partially neutralized surface, DNA neutralized at seven positions to simulate PU.1 binding is observed to bend by 28 degrees . The directions of DNA curvature are slightly different in the co crystal versus the partially neutralized duplexes. The electrostatic component of the binding energy appears more than enough to account for the DNA bending observed in the PU.1-DNA complex. PMID- 9395495 TI - G protein beta1gamma2 subunits promote microtubule assembly. AB - alpha and betagamma subunits of G proteins are thought to transduce signals from cell surface receptors to intracellular effector molecules. Galpha and Gbetagamma have also been implicated in cell growth and differentiation, perhaps due to their association with cytoskeletal components. In this report Gbetagamma is shown to modulate the cytoskeleton by regulation of microtubule assembly. Specificity among betagamma species exists, as beta1gamma2 stimulates microtubule assembly, and beta1gamma1 is without any effect. Furthermore, a mutant beta1gamma2, beta1gamma2(C68S), which does not undergo prenylation and subsequent carboxyl-terminal processing on the gamma subunit, does not stimulate the formation of microtubules. beta immunoreactivity was detected exclusively in the microtubule fraction after assembly in the presence of beta1gamma2, suggesting a preferential association with microtubules rather than soluble tubulin. Crude microtubule fractions from ovine brain contain Gbetagamma, and electron microscopy reveals a specific association with microtubules. The decoration of microtubules by Gbetagamma appears to be strikingly similar to the periodic pattern observed for microtubule-associated proteins, suggesting a similar site of activation of microtubule assembly by both agents. It is suggested that reformation of the cytoskeleton represents an additional cellular process mediated by Gbetagamma. PMID- 9395496 TI - Inhibition of vascular endothelial growth factor (VEGF)-induced endothelial cell proliferation by a peptide corresponding to the exon 7-encoded domain of VEGF165. AB - Vascular endothelial growth factor (VEGF) is a potent mitogen for endothelial cells (EC) in vitro and a major regulator of angiogenesis in vivo. VEGF121 and VEGF165 are the most abundant of the five known VEGF isoforms. The structural difference between these two is the presence in VEGF165 of 44 amino acids encoded by exon 7 lacking in VEGF121. It was previously shown that VEGF165 and VEGF121 both bind to KDR/Flk-1 and Flt-1 but that VEGF165 binds in addition to a novel receptor (Soker, S., Fidder, H., Neufeld, G., and Klagsbrun, M. (1996) J. Biol. Chem. 271, 5761-5767). The binding of VEGF165 to this VEGF165-specific receptor (VEGF165R) is mediated by the exon 7-encoded domain. To investigate the biological role of this domain further, a glutathione S-transferase fusion protein corresponding to the VEGF165 exon 7-encoded domain was prepared. The fusion protein inhibited binding of 125I-VEGF165 to VEGF165R on human umbilical vein-derived EC (HUVEC) and MDA-MB-231 tumor cells. The fusion protein also inhibited significantly 125I-VEGF165 binding to KDR/Flk-1 on HUVEC but not on porcine EC which express KDR/Flk-1 alone. VEGF165 had a 2-fold higher mitogenic activity for HUVEC than did VEGF121. The exon 7 fusion protein inhibited VEGF165 induced HUVEC proliferation by 60% to about the level stimulated by VEGF121. Unexpectedly, the fusion protein also inhibited HUVEC proliferation in response to VEGF121. Deletion analysis revealed that a core inhibitory domain exists within the C-terminal 23-amino acid portion of the exon 7-encoded domain and that a cysteine residue at position 22 in exon 7 is critical for inhibition. It was concluded that the exon 7-encoded domain of VEGF165 enhances its mitogenic activity for HUVEC by interacting with VEGF165R and modulating KDR/Flk-1-mediated mitogenicity indirectly and that exon 7-derived peptides may be useful VEGF antagonists in angiogenesis-associated diseases. PMID- 9395497 TI - MAGI-1, a membrane-associated guanylate kinase with a unique arrangement of protein-protein interaction domains. AB - Membrane-associated guanylate kinase (MAGUK) proteins participate in the assembly of multiprotein complexes on the inner surface of the plasma membrane at regions of cell-cell contact. MAGUKs are characterized by three types of protein-protein interaction modules: the PDZ domain, the Src homology 3 (SH3) domain, and the guanylate kinase (GuK) domain. The arrangement of these domains is conserved in all previously known MAGUKs: either one or three PDZ domains in the NH2-terminal half, followed by the SH3 domain, followed by a COOH-terminal GuK domain. In this report, we describe the cDNA cloning and subcellular distribution of MAGI-1, a MAGUK with three unique structural features: 1) the GuK domain is at the NH2 terminus, 2) the SH3 domain is replaced by two WW domains, and 3) it contains five PDZ domains. MAGI-1 mRNA was detected in several adult mouse tissues. Sequence analysis of overlapping cDNAs revealed the existence of three splice variants that are predicted to encode MAGI-1 proteins with different COOH termini. The longest variant, MAGI-1c, contains three bipartite nuclear localization signals in its unique COOH-terminal sequence and was found predominantly in the nucleus of Madin-Darby canine kidney cells. A shorter form lacking these signals was found primarily in membrane and cytoplasmic fractions. This distribution, which is reminiscent of that seen for the tight junction protein ZO-1, suggests that MAGI-1 may participate in the transmission of regulatory signals from the cell surface to the nucleus. PMID- 9395498 TI - Suppression of neuronal and cardiac transient outward currents by viral gene transfer of dominant-negative Kv4.2 constructs. AB - To probe the molecular identity of transient outward (A-type) potassium currents, we expressed a truncated version of Kv4.2 in heart cells and neurons. The rat Kv4.2-coding sequence was truncated at a position just past the first transmembrane segment and subcloned into an adenoviral shuttle vector downstream of a cytomegalovirus promoter (pE1Kv4.2ST). We hypothesized that this construct would act as a dominant-negative suppressor of currents encoded by the Kv4 family by analogy to Kv1 channels. Cotransfection of wild-type Kv4.2 with a beta galactosidase expression vector in Chinese hamster ovary (CHO)-K1 cells produced robust transient outward currents (Ito) after two days (14.0 pA/pF at 50 mV, n = 5). Cotransfection with pE1Kv4.2ST markedly suppressed the Kv4.2 currents (0.8 pA/pF, n = 6, p < 0.02; cDNA ratio of 2:1 Kv4.2ST:wild type), but in parallel experiments, it did not alter the current density of coexpressed Kv1.4 or Kv1.5 channels. Kv4.2ST also effectively suppressed rat Kv4.3 current when coexpressed in CHO-K1 cells. We then engineered a recombinant adenovirus (AdKv4.2ST) designed to overexpress Kv4.2ST in infected cells. A-type currents in rat cerebellar granule cells were decreased two days after AdKv4. 2ST infection as compared with those infected by a beta-galactosidase reporter virus (116.0 pA/pF versus 281.4 pA/pF in Ad beta-galactosidase cells, n = 8 each group, p < 0.001). Likewise, Ito in adult rat ventricular myocytes was suppressed by AdKv4.2ST but not by Adbeta galactosidase (8.8 pA/pF versus 21.4 pA/pF in beta-galactosidase cells, n = 6 each group, p < 0.05). Expression of a GFP-Kv4.2ST fusion construct enabled imaging of subcellular protein localization by confocal microscopy. The protein was distributed throughout the surface membrane and intracellular membrane systems. We conclude that genes from the Kv4 family are the predominant contributors to the A-type currents in cerebellar granule cells and Ito in rat ventricle. Overexpression of dominant-negative constructs may be of general utility in dissecting the contributions of various ion channel genes to excitability. PMID- 9395499 TI - Functional coupling of NKR-P1 receptors to various heterotrimeric G proteins in rat interleukin-2-activated natural killer cells. AB - NKR-P1 molecules constitute a family of type II membrane receptors in natural killer (NK) cells that preferentially activate NK cell killing and release of interferon-gamma from these cells. Here, we demonstrate that anti-NKR-P1 enhances GTP binding in rat interleukin-2-activated NK cell membranes; GTP binding to Gi3alpha, Gsalpha, Gq,11alpha, and Gzalpha increased noticeably in these cell membranes after treatment with anti-NKR-P1. Western blot analysis of membrane proteins prepared from interleukin-2-activated NK cells reveals the presence of Gi1,2alpha, Gi3alpha, Goalpha, Gsalpha, Gq, 11alpha, Gzalpha, and G12alpha, but not G13alpha. However, only alphai3, alphas, alphaq,11, and alphaz, but not alphai1,2, alphao, alpha12, or alpha13 subunits when immunoprecipitated with the appropriate anti-G protein antibodies, are associated with NKR-P1 when immunoblotted with anti-NKR-P1. Reciprocally, NKR-P1 immunoprecipitated with anti NKR-P1 is associated with alphai3, alphas, alphaq,11, and alphaz immunoblotted with anti-G proteins. These results are the first to demonstrate the physical and functional coupling of NKR-P1 to the heterotrimeric G proteins in NK cells. PMID- 9395500 TI - Selective cytotoxicity of dermaseptin S3 toward intraerythrocytic Plasmodium falciparum and the underlying molecular basis. AB - The antimicrobial activity of various naturally occurring microbicidal peptides was reported to result from their interaction with microbial membrane. In this study, we investigated the cytotoxicity of the hemolytic peptide dermaseptin S4 (DS4) and the nonhemolytic peptide dermaseptin S3 (DS3) toward human erythrocytes infected by the malaria parasite Plasmodium falciparum. Both DS4 and DS3 inhibited the parasite's ability to incorporate [3H]hypoxanthine. However, while DS4 was toxic toward both the parasite and the host erythrocyte, DS3 was toxic only toward the intraerythrocytic parasite. To gain insight into the mechanism of this selective cytotoxicity, we labeled the peptides with fluorescent probes and investigated their organization in solution and in membranes. In Plasmodium infected cells, rhodamine-labeled peptides interacted directly with the intracellular parasite, in contrast to noninfected cells, where the peptides remained bound to the erythrocyte plasma membrane. Binding experiments to phospholipid membranes revealed that DS3 and DS4 had similar binding characteristics. Membrane permeation studies indicated that the peptides were equally potent in permeating phosphatidylserine/phosphatidylcholine vesicles, whereas DS4 was more permeative with phosphatidylcholine vesicles. In aqueous solutions, DS4 was found to be in a higher aggregation state. Nevertheless, both DS3 and DS4 spontaneously dissociated to monomers upon interaction with vesicles, albeit with different kinetics. In light of these results, we propose a mechanism by which dermaseptins permeate cells and affect intraerythrocytic parasites. PMID- 9395501 TI - Heparin-binding properties of the amyloidogenic peptides Abeta and amylin. Dependence on aggregation state and inhibition by Congo red. AB - Aggregation and deposition of the 40-42-residue amyloid beta-protein (Abeta) are early and necessary neuropathological events in Alzheimer's disease. An understanding of the molecular interactions that trigger these events is important for therapeutic strategies aimed at blocking Abeta plaque formation at the earliest stages. Heparan sulfate proteoglycans may play a fundamental role since they are invariably associated with Abeta and other amyloid deposits at all stages. However, the nature of the Abeta-heparan sulfate proteoglycan binding has been difficult to elucidate because of the strong tendency of Abeta to self aggregate. Affinity co-electrophoresis can measure the binding of proteoglycans or glycosaminoglycans to proteins without altering the physical state of the protein during the assay. We used affinity co-electrophoresis to study the interaction between Abeta and the glycosaminoglycan heparin and found that the aggregation state of Abeta governs its heparin-binding properties: heparin binds to fibrillar but not nonfibrillar Abeta. The amyloid binding dye, Congo red, inhibited the interaction in a specific and dose-dependent manner. The "Dutch" mutant AbetaE22Q peptide formed fibrils more readily than wild type Abeta and it also attained a heparin-binding state more readily, but, once formed, mutant and wild type fibrils bound heparin with similar affinities. The heparin-binding ability of aggregated AbetaE22Q was reversible with incubation in a solvent that promotes alpha-helical conformation, further suggesting that conformation of the peptide is important. Studies with another human amyloidogenic protein, amylin, suggested that its heparin-binding properties were also dependent on aggregation state. These results demonstrate the dependence of the Abeta-heparin interaction on the conformation and aggregation state of Abeta rather than primary sequence alone, and suggest methods of interfering with this association. PMID- 9395502 TI - Cloning, expression, and properties of the regulatory subunit of bovine pyruvate dehydrogenase phosphatase. AB - cDNA encoding the regulatory subunit of bovine mitochondrial pyruvate dehydrogenase phosphatase (PDPr) has been cloned. Overlapping cDNA fragments were generated by the polymerase chain reaction from bovine genomic DNA and from cDNA synthesized from bovine poly(A)+ RNA and total RNA. The complete cDNA (2885 base pairs) contains an open reading frame of 2634 nucleotides encoding a putative presequence of 31 amino acid residues and a mature protein of 847 residues with a calculated Mr of 95,656. This value is in agreement with the molecular mass of native PDPr (95,800 +/- 200 Da) determined by matrix-assisted laser desorption ionization mass spectrometry. The mature form of PDPr was expressed in Escherichia coli as a maltose-binding protein fusion, and the recombinant protein was purified to near homogeneity. It exhibited properties characteristic of the native PDPr, including recognition by antibodies against native bovine PDPr, ability to decrease the sensitivity of the catalytic subunit to Mg2+, and reversal of this inhibitory effect by the polyamine spermine. A BLAST search of protein data bases revealed that PDPr is distantly related to the mitochondrial flavoprotein dimethylglycine dehydrogenase, which functions in choline degradation. PMID- 9395503 TI - The type of basal promoter determines the regulated or constitutive mode of transcription in the common control region of the yeast gene pair GCY1/RIO1. AB - The yeast genes, GCY1 and RIO1, are transcribed divergently from the 869-base pair intergenic region. GCY1 is inducible by galactose about 25-fold due to Gal4p binding to a single UASGAL, whereas RIO1 is constitutively expressed. GCY1 has a TATA box obeying the consensus TATAAA, whereas the RIO1 5'-upstream region lacks such a motif. In vitro mutagenesis of the TATA motif of GCY1, on the one hand, and introduction of a TATA-element into the promoter of RIO1, on the other hand, as well as inversion of the intergenic region have revealed that transcription of GCY1 and RIO1 is only regulated by Gal4p when a consensus TATA motif is included in their core promoters but not in its absence. The data imply that only transcription complexes that assemble at a consensus TATA box are compatible with specific transactivators, such as Gal4p. As a result, the adjacent gene is subject to regulated expression. By contrast, if a consensus TATA sequence is absent, the initiation complex does not respond to regulatory transcription factors, and consequently, the respective gene is constitutively transcribed. On the other hand, we show that two blocks of homo-oligomeric (dA.dT) sequences do not function as boundary sequences that might confine regulatory action of Gal4p to GCY1. PMID- 9395504 TI - Hsp110 protects heat-denatured proteins and confers cellular thermoresistance. AB - The 110-kDa heat shock protein (hsp110) has long been recognized as one of the primary heat shock proteins in mammalian cells. It belongs to a recently described protein family that is a significantly diverged subgroup of the hsp70 family and has been found in organisms as diverse as yeast and mammals. We describe here the first analysis of the ability of hsp110 to protect cellular and molecular targets from heat damage. It was observed that the overexpression in vivo of hsp110 conferred substantial heat resistance to both Rat-1 and HeLa cells. In vitro heat denaturation and refolding assays demonstrate that hsp110 is highly efficient in selectively recognizing denatured proteins and maintaining them in a soluble, folding-competent state and is significantly more efficient in performing this function than is hsc70. hsp110-bound proteins can then be refolded by the addition of rabbit reticulocyte lysate or hsc70 and Hdj-1, whereas Hdj-1 does not itself function as a co-chaperone in folding with hsp110. hsp110 is one of the principal molecular chaperones of mammalian cells and represents a newly identified component of the primary protection/repair pathway for denatured proteins and thermotolerance expression in vivo. PMID- 9395505 TI - Negative regulation of the mouse aldolase A gene. A cell cycle-dependent DNA binding activity functions as a silencer of gene transcription. AB - The expression of aldolase A L-type mRNA is increased in growth-arrested mouse NIH3T3 cells and remarkably down-regulated in actively proliferating cells. Treatment of proliferating cells with cycloheximide abolished the down-regulation of L-type mRNA expression, thus indicating that a protein factor acts as repressor in proliferating cells. Transient transfection experiments in NIH3T3 cells showed that a negative regulatory cis-element (NRE) is involved in the modulation of the transcriptional activity of the distal L promoter. The repressor, which is a protein of approximately 97 kDa, binds the murine aldolase A NRE, revealing a much more intense DNA-protein complex in proliferating NIH3T3 cells than in serum-deprived cells. Mutations in the negative regulatory cis element showed that the GA-rich motif is required for protein binding and silencer function. We conclude that the expression of L-type mRNA is modulated by the interaction between a cell cycle-dependent DNA-binding protein and the murine aldolase A NRE. PMID- 9395506 TI - G protein-coupled receptors mediate two functionally distinct pathways of tyrosine phosphorylation in rat 1a fibroblasts. Shc phosphorylation and receptor endocytosis correlate with activation of Erk kinases. AB - The Ras-dependent activation of Erk kinases by G protein-coupled receptors (GPCRs) is thought to involve tyrosine phosphorylation of docking proteins that serve as scaffolds for the plasma membrane recruitment of Ras guanine nucleotide exchange factors, such as the Grb2-mSos complex. We have investigated the role of two GPCR-regulated tyrosine phosphoproteins, p125(FAK) (FAK) and Shc, in the Ras dependent activation of Erk kinases by endogenously expressed GPCRs in Rat 1a fibroblasts. Several lines of evidence suggest that tyrosine phosphorylation of FAK and Shc are independently regulated. The GPCRs for lysophosphatidic acid (LPA), thrombin, and bombesin mediate equivalent increases in FAK tyrosine phosphorylation and FAK-Grb2 association. In contrast, only LPA and thrombin receptors significantly stimulate Shc tyrosine phosphorylation and Shc-Grb2 complex formation. Tyrosine phosphorylation of FAK is pertussis toxin insensitive, can be mimicked by calcium ionophore, and is inhibited by treatment with cytochalasin D, which depolymerizes the actin cytoskeleton. In contrast, tyrosine phosphorylation of Shc is inhibited by pertussis toxin treatment, is not induced by calcium ionophore, and is insensitive to cytochalasin D. In each case, the rapid stimulation of Erk 1/2 correlates with tyrosine phosphorylation of Shc but not of FAK. The dissociation of FAK-Grb2 complex formation from receptor mediated activation of Erk 1/2 indicates that recruitment of Grb2-mSos to the plasma membrane is not sufficient to mediate rapid Erk activation. Using four mechanistically distinct inhibitors of clathrin-mediated endocytosis, concanavalin A, hypertonic medium, depletion of intracellular potassium, and monodansylcadaverine, we find that GPCR-mediated Erk 1/2 activation is also endocytosis-dependent. Thus, we propose that an additional step involving vesicle mediated endocytosis is required for the rapid, Ras-dependent activation of Erk kinases in fibroblasts. PMID- 9395507 TI - hsp27 as a switch between differentiation and apoptosis in murine embryonic stem cells. AB - Small stress proteins are developmentally regulated and linked to cell growth and differentiation. The early phase of murine embryonic stem (ES) cell differentiation, characterized by a gradual growth arrest, is accompanied with hsp27 transient accumulation. This differentiation process also correlated with changes in hsp27 phosphorylation and oligomerization. The role of hsp27 was investigated in ES clones stably transfected with murine or human hsp27 genes, placed in sense or antisense orientation. Several clones were obtained that either underexpressed endogenous murine hsp27 or overexpressed murine or human hsp27. Maintained undifferentiated, these clones showed similar growth rates. We report here that hsp27 constitutive overexpression enhanced the differentiation mediated decreased rate of ES cell proliferation but did not alter morphological changes. In contrast, hsp27 underexpression, which attenuated cell growth arrest, induced differentiation abortion because of an overall cell death by apoptosis. Recently, we showed that hsp27 interfered with cell death probably because of its ability to modulate intracellular glutathione. hsp27 accumulation during ES cell differentiation was also correlated with an increase in glutathione, which was attenuated by hsp27 down-expression. Hence, hsp27 transient expression seems essential for preventing differentiating ES cells from undergoing apoptosis, a switch that may be redox regulated. PMID- 9395508 TI - Structural analyses of gp45 sliding clamp interactions during assembly of the bacteriophage T4 DNA polymerase holoenzyme. I. Conformational changes within the gp44/62-gp45-ATP complex during clamp loading. AB - A multisubunit ring-shaped protein complex is used to tether the polymerase to the DNA at the primer-template junction in most DNA replication systems. This "sliding clamp" interacts with the polymerase, completely encircles the DNA duplex, and is assembled onto the DNA by a specific clamp loading complex in an ATP-driven process. Site-specific mutagenesis has been used to introduce single cysteine residues as reactive sites for adduct formation within each of the three subunits of the bacteriophage T4-coded sliding clamp complex (gp45). Two such mutants, gp45S19C and gp45K81C, are reacted with the cysteine-specific photoactivable cross-linker TFPAM-3 and used to track the changes in the relative positioning of the gp45 subunits with one another and with the other components of the clamp loading complex (gp44/62) in the various stages of the loading process. Cross-linking interactions performed in the presence of nucleotide cofactors show that ATP binding and hydrolysis, interaction with primer-template DNA, and release of ADP all result in significant conformational changes within the clamp loading cycle. A structural model is presented to account for the observed rearrangements of intersubunit contacts within the complex during the loading process. PMID- 9395509 TI - Structural analyses of gp45 sliding clamp interactions during assembly of the bacteriophage T4 DNA polymerase holoenzyme. II. The Gp44/62 clamp loader interacts with a single defined face of the sliding clamp ring. AB - The phage T4 gp45 sliding clamp is a ring-shaped replication accessory protein that is mounted onto DNA in an ATP-dependent manner by the gp44/62 clamp loader. In the preceding paper (Pietroni, P., Young, M. C., Latham, G. J., and von Hippel, P. H. (1997) J. Biol. Chem. 272, 31666-31676), two gp45 mutants were exploited to probe interactions of the sliding clamp ring with the gp44/62 loading machinery at various steps during the clamp loading process. In this report, these studies are extended to examine the polarity of the binding interaction between gp45 and gp44/62. Three different gp45 mutants containing a single cysteine in three topographically distinct positions were used. Several different reporter groups, including extrinsic fluorophores, a photo-cross linker, and a biotin linker for use in a novel "streptavidin interference assay," were covalently attached to these cysteine residues. Since gp45 is a trimeric protein, these three different mutations allowed us to probe up to nine distinct local environments along the surface of the sliding clamp in the presence and absence of other replication proteins. The results show that the gp44/62-ATP clamp loader complex binds exclusively to the C-terminal (S19C) face of the gp45 ring. PMID- 9395510 TI - Structural analyses of gp45 sliding clamp interactions during assembly of the bacteriophage T4 DNA polymerase holoenzyme. III. The Gp43 DNA polymerase binds to the same face of the sliding clamp as the clamp loader. AB - In the preceding paper (Latham, G. J., Bacheller, D. J., Pietroni, P. , and von Hippel, P. H. (1997) J. Biol. Chem. 272, 31677-31684), we demonstrated that the T4 gp44/62-ATP clamp loader binds to the C-terminal face of the gp45 sliding clamp. Here we extend these results by exploring the structural relationship between the gp43 polymerase and the gp45 sliding clamp. Using fluorescence intensity and polarization techniques, as well as photo-cross-linking methods, we present evidence that gp43, like gp44/62, binds to the C-terminal face of gp45. In addition, we show that g43 binds to the gp45 clamp in two distinct interaction modes, depending on the presence or absence of template-primer DNA. When template primer DNA is present, gp43 binds tightly to gp45 to form the highly processive DNA polymerase holoenzyme. Gp43 also binds to gp45 in the absence of template primer DNA, but this interaction is more than 100 times weaker than gp43-gp45 binding on DNA. Specific interactions between gp43 and the C-terminal face of gp45 are maintained in both modes of binding. These results underscore the pivotal role of template-primer DNA in modulating the strength of protein-protein interactions during DNA synthesis and provide additional insight into the structural requirements of the replication process. PMID- 9395511 TI - A role for estrogen-related receptor alpha in the control of mitochondrial fatty acid beta-oxidation during brown adipocyte differentiation. AB - Little is known about the factors involved in the brown adipocyte gene regulatory program. In contrast to the white adipocyte, the brown adipocyte is characterized by abundant mitochondria and high level expression of mitochondrial fatty acid beta-oxidation enzymes. Previous studies in transgenic mice have shown that the brown adipose-enriched expression of a key beta-oxidation enzyme, medium chain acyl-coenzyme A dehydrogenase (MCAD), requires cis-acting elements located within the proximal promoter region of the MCAD gene. The levels of mRNA encoding MCAD and several other beta-oxidation cycle enzymes were coordinately induced during differentiation of brown adipocytes in culture. Expression of transgenes comprised of MCAD gene promoter fragments fused to chloramphenicol acetyltransferase reporters in differentiating brown adipocytes revealed that a known nuclear receptor response element (NRRE-1) was required for the transcriptional induction of the MCAD gene during brown adipocyte differentiation. Electrophoretic mobility shift assays and antibody recognition studies identified distinct brown adipocyte differentiation stage-specific, NRRE 1-protein complexes; the orphan nuclear receptors, chicken ovalbumin upstream promoter transcription factors I and II, were identified as major the NRRE-1 binding proteins in the pre-adipocyte, whereas the estrogen-related receptor alpha (ERRalpha) bound NRRE-1 in extracts prepared from differentiated brown adipocytes. DNA binding studies performed with a series of NRRE-1 mutant probes indicated that ERRalpha was capable of binding two distinct sites within NRRE-1, each of which conform to the known ERRalpha monomeric binding consensus. The expression of ERRalpha paralleled NRRE-1 binding activities and MCAD expression during brown adipocyte differentiation, cardiac development, and among a variety of adult mouse tissues. These results identify a new class of ERRalpha target genes and implicate ERRalpha and chicken ovalbumin upstream promoter transcription factor in the control of a pivotal metabolic pathway during brown adipocyte differentiation. PMID- 9395512 TI - Aggregated low density lipoprotein induces and enters surface-connected compartments of human monocyte-macrophages. Uptake occurs independently of the low density lipoprotein receptor. AB - Aggregation of low density lipoprotein (LDL) stimulates its uptake by macrophages. We have now shown by electron microscopic and chemical experiments that aggregated LDL (produced by vortexing (VxLDL) or treatment with phospholipase C) induced and became sequestered in large amounts within surface connected compartments (SCC) of human monocyte-derived macrophages. This occurred through a process different from phagocytosis. Formation of SCC and accumulation of aggregated LDL in SCC are cell-mediated processes that were temperature dependent (10 x greater cell association at 37 degrees C than at 4 degrees C) and blocked by cytochalasin D but not by nocodazole. Because of the surface connections of SCC, trypsin could release aggregated LDL from SCC. Degradation of 125I-VxLDL through the SCC pathway showed delayed and a lower rate of degradation (10-55%) compared with nonaggregated 125I-acetylated LDL that did not enter SCC. However, similar to 125I-acetylated LDL degradation, 125I-VxLDL degradation occurred through a chloroquine-sensitive pathway. Uptake of VxLDL into SCC was not mediated by the LDL receptor. Methylation of LDL prevents its binding to the LDL receptor. However, methylated LDL still entered SCC after it was aggregated by vortexing. On the other hand, degradation of 125I-VxLDL was substantially decreased by methylation of LDL and by cholesterol enrichment of macrophages, which decreases macrophage LDL receptor expression. The results suggest that whereas uptake of aggregated LDL into SCC occurs independently of the LDL receptor, movement of aggregated LDL from SCC to lysosomes may depend in part on LDL receptor function. Sequestration into SCC is a novel endocytosis pathway for uptake of aggregated LDL that allows the macrophage to store large amounts of this lipoprotein before it is further processed. PMID- 9395513 TI - Furin-mediated cleavage of Pseudomonas exotoxin-derived chimeric toxins. AB - Pseudomonas exotoxin (PE) requires proteolytic cleavage to generate a 37-kDa C terminal fragment that translocates to the cytosol and ADP-ribosylates elongation factor 2. Cleavage within cells is mediated by furin, occurs between arginine 279 and glycine 280, and requires an arginine at both P1 and P4 residues. To study the proteolytic processing of PE-derived chimeric toxins, TGFalpha-PE38 (transforming growth factor fused to the domains II and III of PE) and a mutant form, TGFalpha-PE38gly279, were each produced in Escherichia coli. When assessed on various epidermal growth factor (EGF) receptor-positive cell lines, TGFalpha PE38 was 100-500-fold more toxic than TGFalpha-PE38gly279. In contrast to PE, where cleavage by furin is only evident at pH 5.5, furin cleaved TGFalpha-PE38 over a broad pH range, while TGFalpha-PE38gly279 was resistant to cleavage. TGFalpha-PE38 was poorly toxic for furin-deficient LoVo cells, unless it was first pretreated in vitro with furin. Furin treatment produced a nicked protein that was 30-fold more toxic than its unnicked counterpart. Using the single chain immunotoxin HB21scFv-PE40 as a substrate, furin-mediated processing of an antibody-based immunotoxin was also evaluated. HB21scFv-PE40, which targets cells expressing the transferrin receptor, was cleaved in a similar fashion to that of TGFalpha-PE38 and nicked HB21scFv-PE40 exhibited increased toxicity for LoVo cells. In short-term experiments, the rate of reduction in protein synthesis by furin-nicked immunotoxins was increased compared with unnicked protein, indicating that cleavage by furin can be a rate-limiting step. We conclude that furin-mediated cleavage of PE-derived immunotoxins is important for their cytotoxic activity. PMID- 9395514 TI - Specific interaction of eukaryotic translation initiation factor 5 (eIF5) with the beta-subunit of eIF2. AB - Eukaryotic translation initiation factor 5 (eIF5) interacts with the 40 S initiation complex (40 S.mRNA. eIF3.Met-tRNAf.eIF2.GTP) and mediates hydrolysis of the bound GTP. To characterize the molecular interactions involved in eIF5 function, we have used 32P-labeled recombinant rat eIF5 as a probe in filter overlay assay to identify eIF5-interacting proteins in crude initiation factor preparations. We observed that eIF5 specifically interacted with the beta subunit of initiation factor eIF2. No other initiation factors including the gamma subunit of eIF2 tested positive in this assay. Furthermore, both yeast and mammalian eIF5 bind to the beta subunit of either mammalian or yeast eIF2. Binding analysis with human eIF2beta deletion mutants expressed in Escherichia coli identified a 22-amino acid domain, between amino acids 68 and 89, as the primary eIF5-binding region of eIF2beta. These results along with our earlier observations that (a) eIF5 neither binds nor hydrolyzes free GTP or GTP bound as Met-tRNAf.eIF2.GTP ternary complex, and (b) eIF5 forms a specific complex with eIF2 suggests that the specific interaction between eIF5 and the beta subunit of eIF2 may be critical for the hydrolysis of GTP during translation initiation. PMID- 9395516 TI - Inhibition of 3-hydroxy-3-methylglutaryl (HMG)-CoA reductase blocks hypoxia mediated down-regulation of endothelial nitric oxide synthase. AB - Hypoxia induces vasoconstriction, in part, by down-regulating endothelial cell nitric oxide synthase (ecNOS) expression. Previous studies indicate that 3 hydroxy-3-methylglutaryl-coenzyme A (HMG CoA) reductase inhibitors improve endothelium-dependent relaxation by increasing ecNOS activity. To determine whether HMG CoA reductase inhibitors can prevent hypoxia-mediated down-regulation of ecNOS function and expression, human endothelial cells were exposed to hypoxia (3% O2) in the presence of HMG CoA reductase inhibitors simvastatin and lovastatin for various durations (0-48 h). Hypoxia decreased ecNOS protein and mRNA levels in a time-dependent manner, resulting in a 4- and 9-fold reduction after 48 h, respectively. In a concentration-dependent manner, simvastatin, and to a lesser extent, lovastatin, prevented the down-regulation of ecNOS expression by hypoxia. Simvastatin-induced changes in ecNOS expression correlated with changes in endothelial NO production and were reversed by treatment with L mevalonate. Actinomycin D studies revealed that under hypoxic conditions, simvastatin increased ecNOS mRNA half-life from 13 to 38 h. Nuclear run-on studies showed that simvastatin had no effect on repression of ecNOS gene transcription by hypoxia. These results indicate that HMG CoA reductase inhibitors regulate ecNOS function and expression through changes in ecNOS mRNA stability and suggest that treatment with HMG CoA reductase inhibitors may have beneficial effects in patients with hypoxia-mediated pulmonary hypertension. PMID- 9395515 TI - Nucleotide sequence context effect of a cyclobutane pyrimidine dimer upon RNA polymerase II transcription. AB - We have studied the role of sequence context upon RNA polymerase II arrest by a cyclobutane pyrimidine dimer using an in vitro transcription system consisting of templates containing a specifically located cyclobutane pyrimidine dimer (CPD) and purified RNA polymerase II (RNAP II) and initiation factors. We selected a model sequence containing a well characterized site for RNAP II arrest in vitro, the human histone H3.3 gene arrest site. The 13-base pair core of the arrest sequence contains two runs of T in the nontranscribed strand that impose a bend in the DNA. We hypothesized that arrest of RNAP II might be affected by the presence of a CPD, based upon the observation that a CPD located at the center of a dA6.dT6 tract eliminates bending (Wang, C.-I., and Taylor, J.-S. (1991) Proc. Natl. Acad. Sci. U. S. A. 88, 9072-9076). We examined the normal H3.3 sequence and a mutant sequence containing a T --> G transversion, which reduces bending and efficiency of arrest. We show that a CPD in the transcribed strand at either of two locations in the arrest site is a potent block to transcription. However, a CPD in the nontranscribed strand only transiently pauses RNAP II. The CPD in concert with a mutation in the arrest site can reduce the extent of bending of the DNA and improve readthrough efficiency. These results demonstrate the potential importance of sequence context for the effect of CPDs within transcribed sequences. PMID- 9395517 TI - Matrix metalloproteinase-3 releases active heparin-binding EGF-like growth factor by cleavage at a specific juxtamembrane site. AB - Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is synthesized as a membrane-anchored precursor that is cleaved to release the soluble mature growth factor. The two forms are active as juxtacrine and paracrine/autocrine growth factors, respectively. The enzymes that process the HB EGF transmembrane form are unknown. Accordingly, an in vitro assay was established using a fusion protein in which alkaline phosphatase (AP) replaced the transmembrane and cytoplasmic domains of HB-EGF (HB-EGF JM-AP). The fusion protein was anchored to agarose beads coated with anti-AP antibodies. Several matrix metalloproteinases (MMPs) were tested for the ability to release soluble HB-EGF in the in vitro system. MMP-3 released soluble 12-kDa immunoreactive and mitogenic HB-EGF within 30 min. On the other hand neither MMP-2 nor MMP-9 had any cleavage activities. A non-cleavable mutant was prepared by replacing the juxtamembrane (JM) region of HB-EGF with the JM region of CD4. The mutant HB-EGF, which in its full-length form was as active a juxtacrine growth factor as was the wild type HB-EGF in vivo, was not cleaved by MMP-3 in the in vitro assay. The C terminal portion of the cleaved HB-EGF JM-AP that remained attached to the anti AP beads was N-terminally sequenced and the MMP-3 cleavage site was determined to be Glu151-Asn152, a site within the JM domain. MMP-3 treatment also released soluble HB-EGF in vivo from MC2 cells expressing transmembrane HB-EGF precursor, at a level of about 2-fold above control. It was concluded that MMP-3 cleaves HB EGF at a specific site in the JM domain and that this enzyme might regulate the conversion of HB-EGF from being a juxtacrine to a paracrine/autocrine growth factor. PMID- 9395518 TI - Cig30, a mouse member of a novel membrane protein gene family, is involved in the recruitment of brown adipose tissue. AB - We have identified a previously uncharacterized gene that is implicated in the thermogenic function of brown adipose tissue of mice. This gene, termed Cig30, is the first mammalian member of a novel gene family comprising several nematode and yeast genes, such as SUR4 and FEN1, mutation of which is associated with highly pleiotropic phenotypes. It codes for a 30-kDa plasma membrane glycoprotein with five putative transmembrane domains. The Cig30 mRNA was readily detected only in brown fat and liver. When animals were exposed to a 3-day cold stress, the Cig30 expression was selectively elevated in brown fat more than 200-fold. Similar increases were brought about in two other conditions of brown fat recruitment, namely during perinatal development and after cafeteria diet. The magnitude of Cig30 mRNA induction in the cold could be mimicked by chronic norepinephrine treatment in vivo. However, in primary cultures of brown adipocytes, a synergistic action of norepinephrine and dexamethasone was required for full expression of the gene, indicating that both catecholamines and glucocorticoids are required for the induction of Cig30. We propose that the CIG30 protein is involved in a pathway connected with brown fat hyperplasia. PMID- 9395519 TI - Ccs1, a nuclear gene required for the post-translational assembly of chloroplast c-type cytochromes. AB - Nuclear genes play important regulatory roles in the biogenesis of the photosynthetic apparatus of eukaryotic cells by encoding factors that control steps ranging from chloroplast gene transcription to post-translational processes. However, the identities of these genes and the mechanisms by which they govern these processes are largely unknown. By using glass bead-mediated transformation to generate insertional mutations in the nuclear genome of Chlamydomonas reinhardtii, we have generated four mutants that are defective in the accumulation of the cytochrome b6f complex. One of them, strain abf3, also fails to accumulate holocytochrome c6. We have isolated a gene, Ccs1, from a C. reinhardtii genomic library that complements both the cytochrome b6f and cytochrome c6 deficiencies in abf3. The predicted protein product displays significant identity with Ycf44 from the brown alga Odontella sinensis, the red alga Porphyra purpurea, and the cyanobacterium Synechocystis strain PCC 6803 (25 33% identity). In addition, we note limited sequence similarity with ResB of Bacillus subtilis and an open reading frame in a homologous operon in Mycobacterium leprae (11-12% identity). On the basis of the pleiotropic c-type cytochrome deficiency in the ccs1 mutant, the predicted plastid localization of the protein, and its relationship to candidate cytochrome biosynthesis proteins in Gram-positive bacteria, we conclude that Ccs1 encodes a protein that is required for chloroplast c-type holocytochrome formation. PMID- 9395520 TI - Signal transduction-mediated activation of the aryl hydrocarbon receptor in rat hepatoma H4IIE cells. AB - We have investigated mechanisms of omeprazole (OME)-mediated induction of CYP1A1 and CYP3A, using the rat hepatoma H4IIE cell line, in comparison with mechanisms exerted by traditional aryl hydrocarbon receptor (AhR) ligands such as benso(a)pyrene (B(a)P) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). OME did not bind specifically to AhR, and it could not activate the AhR complex in rat cytosol to a xenobiotic-responsive element (XRE)-binding form in vitro. Genistein, a tyrosine kinase inhibitor, and daidzein, an inhibitor of casein kinase II, efficiently inhibited OME-mediated but not B(a)P- or TCDD-mediated induction of CYP1A1, as monitored at the transcriptional, mRNA, and protein levels as well as by analysis of activation of XRE-luciferase reporter constructs transfected into H4IIE cells. The protease inhibitor Nalpha-p-tosyl-L-lysine chloromethyl ketone (TLCK) and lavendustin A also had similar OME-specific effects. In addition, insulin pretreatment caused an almost complete inhibition of OME-dependent CYP1A1 induction but only partially affected TCDD and B(a)P mediated induction of CYP1A1. Staurosporine, an inhibitor of protein kinase C, impaired the induction by both B(a)P and OME. OME caused an approximately 2-fold increase in the level of CYP3A expression, but all inhibitors used were ineffective in preventing this induction. Gel shift analysis with radiolabeled XRE and specific peptide antibodies toward AhR and aryl hydrocarbon receptor nuclear translocator protein (Arnt) revealed an OME-mediated translocation of the AhR.Arnt complex into the nuclei. Genistein inhibited the specific nuclear XRE binding caused by OME, but it potentiated the formation of the TCDD-induced XRE.AhR complex. Although daidzein was able to effectively inhibit the OME stimulated CYP1A1 gene transcription, it did not influence the OME-dependent AhR.XRE complex formation. The data are consistent with a mechanism for OME mediated induction of CYP1A1 that involves activation of the AhR complex via intracellular signal transduction systems and that is distinct from induction mediated by AhR ligands. PMID- 9395521 TI - Role of metalloproteases in the release of the IL-1 type II decoy receptor. AB - The IL-1 type II receptor (decoy RII) is a nonsignaling molecule the only established function of which is to capture IL-1 and prevent it from interacting with signaling receptor. The decoy RII is released in a regulated way from the cell surface. Here, we reported that hydroxamic acid inhibitors of matrix metalloproteases inhibit different pathways of decoy RII release, including the following: (a) the slow (18 h) gene expression-dependent release from monocytes and polymorphonuclear cells exposed to dexamethasone; (b) rapid release (minutes) from myelomonocytic cells exposed to tumor necrosis factor, chemoattractants, or phorbol myristate acetate; (c) phorbol myristate acetate-induced release from decoy RII-transfected fibroblasts and B cells. Inhibition of release was associated with increased surface expression of decoy RII. Inhibitors of other protease classes did not substantially affect release. However, serine protease inhibitors increased the molecular mass of the decoy RII released from polymorphonuclear cells from 45 to 60 kDa. Thus, irrespective of the pathway responsible for release and of the cellular context, matrix metalloproteases, rather than differential splicing, play a key role in production of soluble decoy RII. PMID- 9395522 TI - Characterization and purification of human corneodesmosin, an epidermal basic glycoprotein associated with corneocyte-specific modified desmosomes. AB - Using monoclonal antibodies, we identified a new protein in mammalian epidermis, which we called corneodesmosin. It is located in the extracellular part of the modified desmosomes in the cornified layer of the tissue, and its proteolysis (from 52-56 to 33 kDa) is thought to be a major prerequisite of desquamation. We have now further characterized human corneodesmosin. Proteolysis of purified cornified cell envelopes produced immunoreactive fragments, confirming the covalent linkage of the protein to these structures. Sequential extraction of epidermal proteins indicated that the 52-56-kDa precursor form of the protein exists in two distinct pools, one extracted with a nondenaturing hypotonic buffer, and the other with urea. Two-dimensional gel analysis and reactivity with phosphoserine-specific antibodies showed that it is a basic phosphoprotein. Deglycosylation experiments, reactivity with lectins, and chromatography on concanavalin A-Sepharose indicated that corneodesmosin is N-glycosylated. Partial sequences, 10 and 15 amino acids long, of the purified 52-56-kDa corneodesmosin showed identity with sequences predicted from a previously cloned gene, proved to be expressed in the epidermis and designated S. This indicates that corneodesmosin is probably encoded by the S gene, the function of which was unknown until now. A model of corneodesmosin maturation during cornification is proposed. PMID- 9395523 TI - A transcription activator with restricted tissue distribution regulates cell specific expression of alpha1(XI) collagen. AB - Different regulatory programs are likely to control expression of the alpha1(XI) collagen (COL11A1) gene in cartilaginous and non-cartilaginous tissues and in coordination with different collagen genes. Here, we report the identification of a cis-acting element that is required for constitutive and tissue-specific activity of the proximal COL11A1 promoter. The element binds an apparently novel activator whose expression is restricted mostly, but not exclusively, to cells of mesenchymal origin. Transient transfection experiments using wild-type and mutant constructs demonstrated the critical contribution of a 45-base pair upstream element (FP9) to promoter activity. The same functional tests and DNA binding assays narrowed down the critical portion of FP9 to a 20-base pair sequence, which consists of an imperfect palindrome with strong homology to the GATA consensus motif. Despite being able to bind GATA proteins in vitro, FP9 is actually recognized by a distinct approximately 100-kDa polypeptide (FP9C) probably belonging to the zinc-finger family of transcription factors. FP9C binding was mostly identified in nuclei from cells of mesenchymal origin, including those actively engaged in COL11A1 transcription. A positive correlation was also established between the level of FP9C binding and the degree of cell differentiation in vitro. Thus, FP9C represents an unusual example of tissue specific and differentiation-related transcription factor with overlapping expression in hard and soft connective tissues. PMID- 9395524 TI - Platelet factor 4 binds to glycanated forms of thrombomodulin and to protein C. A potential mechanism for enhancing generation of activated protein C. AB - Platelet factor 4 (PF4) is an abundant platelet alpha-granule heparin-binding protein. We have previously shown that PF4 accelerates up to 25-fold the proteolytic conversion of protein C to activated protein C by the thrombin.thrombomodulin complex by increasing its affinity for protein C 30-fold. This stimulatory effect requires presence of the gamma-carboxyglutamic acid (Gla) domain in protein C and is enhanced by the presence of a chondroitin sulfate glycosaminoglycan (GAG) domain on thrombomodulin. We hypothesized that cationic PF4 binds to both protein C and thrombomodulin through these anionic domains. Qualitative SDS-polyacrylamide gel electrophoresis analysis of avidin extracts of solutions containing biotinylated PF4 and candidate ligands shows that PF4 binds to GAG+ but not GAG- forms of thrombomodulin and native but not Gla-domainless protein C. Quantitative analysis using the surface plasmon resonance-based BIAcoreTM biosensor system confirms the extremely high affinity of PF4 for heparin (KD = 4 nM) and shows that PF4 binds to GAG+ thrombomodulin with a KD of 31 nM and to protein C with a KD of 0.37 microM. In contrast, PF4 had no measurable interaction with GAG- thrombomodulin or Gla-domainless protein C. Western blot analysis of normal human plasma extracted with biotinylated PF4 demonstrates PF4 binding to protein C in a physiologic context. Thus, PF4 binds with relative specificity and high affinity to the GAG- domain of thrombomodulin and the Gla domain of protein C. These interactions may enhance the affinity of the thrombin.thrombomodulin complex for protein C and thereby promote the generation of activated protein C. PMID- 9395525 TI - CCAAT/enhancer-binding protein delta activates insulin-like growth factor-I gene transcription in osteoblasts. Identification of a novel cyclic AMP signaling pathway in bone. AB - Insulin-like growth factor-I (IGF-I) plays a key role in skeletal growth by stimulating bone cell replication and differentiation. We previously showed that prostaglandin E2 (PGE2) and other cAMP-activating agents enhanced IGF-I gene transcription in cultured primary rat osteoblasts through promoter 1, the major IGF-I promoter, and identified a short segment of the promoter, termed HS3D, that was essential for hormonal regulation of IGF-I gene expression. We now demonstrate that CCAAT/enhancer-binding protein (C/EBP) delta is a major component of a PGE2-stimulated DNA-protein complex involving HS3D and find that C/EBPdelta transactivates IGF-I promoter 1 through this site. Competition gel shift studies first indicated that a core C/EBP half-site (GCAAT) was required for binding of a labeled HS3D oligomer to osteoblast nuclear proteins. Southwestern blotting and UV-cross-linking studies showed that the HS3D probe recognized a approximately 35-kDa nuclear protein, and antibody supershift assays indicated that C/EBPdelta comprised most of the PGE2-activated gel-shifted complex. C/EBPdelta was detected by Western immunoblotting in osteoblast nuclear extracts after treatment of cells with PGE2. An HS3D oligonucleotide competed effectively with a high affinity C/EBP site from the rat albumin gene for binding to osteoblast nuclear proteins. Co-transfection of osteoblast cell cultures with a C/EBPdelta expression plasmid enhanced basal and PGE2-activated IGF-I promoter 1-luciferase activity but did not stimulate a reporter gene lacking an HS3D site. By contrast, an expression plasmid for the related protein, C/EBPbeta, did not alter basal IGF-I gene activity but did increase the response to PGE2. In osteoblasts and in COS-7 cells, C/EBPdelta, but not C/EBPbeta, transactivated a reporter gene containing four tandem copies of HS3D fused to a minimal promoter; neither transcription factor stimulated a gene with four copies of an HS3D mutant that was unable to bind osteoblast nuclear proteins. These results identify C/EBPdelta as a hormonally activated inducer of IGF-I gene transcription in osteoblasts and show that the HS3D element within IGF-I promoter 1 is a high affinity binding site for this protein. PMID- 9395526 TI - The recycling of ERGIC-53 in the early secretory pathway. ERGIC-53 carries a cytosolic endoplasmic reticulum-exit determinant interacting with COPII. AB - Further investigation of the targeting of the intracellular membrane lectin endoplasmic reticulum (ER)-Golgi intermediate compartment-53 (ERGIC-53) by site directed mutagenesis revealed that its lumenal and transmembrane domains together confer ER retention. In addition we show that the cytoplasmic domain is required for exit from the ER indicating that ERGIC-53 carries an ER-exit determinant. Two phenylalanines at the C terminus are essential for ER-exit. Thus, ERGIC-53 contains determinants for ER retention as well as anterograde transport which, in conjunction with a dilysine ER retrieval signal, control the continuous recycling of ERGIC-53 in the early secretory pathway. In vitro binding studies revealed a specific phenylalanine-dependent interaction between an ERGIC-53 cytosolic tail peptide and the COPII coat component Sec23p. These results suggest that the ER exit of ERGIC-53 is mediated by direct interaction of its cytosolic tail with the Sec23p.Sec24p complex of COPII and that protein sorting at the level of the ER occurs by a mechanism similar to receptor-mediated endocytosis or Golgi to ER retrograde transport. PMID- 9395527 TI - The Ca2+/calmodulin-dependent kinase type IV is involved in the CD5-mediated signaling pathway in human T lymphocytes. AB - The CD5 receptor on T lymphocytes is involved in T cell activation and T-B cell interactions. In the present study, we have characterized the signaling pathways induced by anti-CD5 stimulation in human T lymphocytes. In T lymphocytes, anti CD5 co-stimulation enhances the phytohemagglutinin/anti-CD28-induced interleukin 2 (IL-2) mRNA accumulation 1.6-fold and IL-2 protein secretion 2. 2-fold, whereby the up-regulation is mediated at both the transcriptional and post transcriptional level. The CD5 signaling pathway up-regulates the IL-2 gene expression by increasing the DNA binding and transactivation activity of activator protein 1 but affects none of the other transcription factors like nuclear factor of activated T cells, nuclear factor kappaB, Oct, and CD28 responsive complex/nuclear factor of mitogen-activated T cells involved in the regulation of the IL-2 promoter activity. The CD5-induced increase of the activator protein 1 activity is mediated through the activation of calcium/calmodulin-dependent (CaM) kinase type IV, and is independent of the activation of mitogen-activated protein kinases Jun N-terminal kinase, extracellular signal-regulated kinase, and p38/Mpk2, and calcium/calmodul independent kinase type II. The expression of a dominant negative mutant of CaM kinase IV in T lymphocytes transfected with an IL-2 promoter-driven reporter construct completely abrogates the response to CD5 stimulation, indicating that CaM kinase IV is essential to the CD5 signaling pathway. In addition, it is demonstrated that calcium/calmodulin-dependent kinase type IV is also involved in the stabilization of the IL-2 transcripts, which is observed after co-stimulation of phytohemagglutinin/anti-CD28 activated T lymphocytes with anti-CD5. PMID- 9395528 TI - Interaction of STAT5 dimers on two low affinity binding sites mediates interleukin 2 (IL-2) stimulation of IL-2 receptor alpha gene transcription. AB - Stimulation of the interleukin 2 receptor alpha (IL-2Ralpha) gene by IL-2 is important for the proliferation of antigen-activated T lymphocytes. IL-2 regulates IL-2Ralpha transcription via a conserved 51-nucleotide IL-2 responsive enhancer. Mouse enhancer function depends on cooperative activity of three distinct sites. Two of these are weak binding sites for IL-2-activated STAT5 (signal transducer and activator of transcription) proteins, and mutational analysis indicates that binding of STAT5 to both sites is required for IL-2 responsiveness of the enhancer. The STAT5 dimers interact to form a STAT5 tetramer. The efficiency of tetramerization depends on the relative rotational orientation of the two STAT motifs on the DNA helix. STAT5 tetramerization on enhancer mutants correlates well with the IL-2 responsiveness of these mutants. This provides strong evidence that interactions between STAT dimers binding to a pair of weak binding sites play a biological role by controlling the activity of a well characterized, complex cytokine-responsive enhancer. PMID- 9395529 TI - Differential ubiquitin-dependent degradation of the yeast apo-cytochrome c isozymes. AB - The yeast Saccharomyces cerevisiae contains two forms of cytochrome c, iso-1- and iso-2-cytochrome c, which are encoded by the nuclear genes CYC1 and CYC7, respectively. The cytochromes c are synthesized in the cytosol, imported into mitochondria, and subsequently modified by the covalent attachment of heme through the action of cytochrome c heme lyase, which is encoded by CYC3. Apo-iso 2-cytochrome c but not apo-iso-1-cytochrome c was observed in cyc3(-) mutants. Furthermore, pulse-chase experiments previously demonstrated that the lack of apo iso-1-cytochrome c was due to its rapid degradation. We report herein that this degradation of apo-iso-1-cytochrome c is dependent on ubiquitination and on the action of the proteasome. Diminished degradation of apo-iso-1-cytochrome c was observed in pre2-2 and pre1-1 mutants having altered proteasome subunits; in ubc1, ubc4, and ubc5 strains lacking one or more of the ubiquitin-conjugating enzymes; and in strains blocked in multi-ubiquitination by overproduction of the abnormal ubiquitin-K48R ubiquitin. In addition, we have used epitope-tagged ubiquitin to demonstrate that apo-iso-1-cytochrome c but not apo-iso-2-cytochrome c is ubiquitinated. Furthermore, the degradation of apo-iso-1-cytochrome c was diminished when the N-terminal region was replaced with the N-terminal region of apo-iso-2-cytochrome c, indicating that this region may be the target for degradation. We suggest that ubiquitin-dependent degradation of apo-iso-1 cytochrome c is part of the regulatory process controlling the preferential expression of the iso-cytochromes c. PMID- 9395530 TI - Variant exons v6 and v7 together expand the repertoire of glycosaminoglycans bound by CD44. AB - Isoforms of the glycoprotein CD44 are cell surface receptors for the glycosaminoglycan hyaluronate. They have been implicated in many biological processes, but their function in these is poorly understood and cannot be explained solely by hyaluronate binding. In the present work we examine the ligand binding properties of alternatively spliced CD44 variant isoforms which are functionally involved in the immune system, embryonic development, and tumor behavior. We show that these isoforms bind directly to the purified glycosaminoglycans chondroitin sulfate, heparin, and heparin sulfate, in addition to being able to bind to hyaluronate. Binding to this extended repertoire of glycosaminoglycans by CD44 depends on the inclusion of peptide sequences encoded by the alternatively spliced exons v6 and v7, and occurs both when the CD44 is solubilized from the plasma membrane and when it is expressed on intact cells. A single point mutation in the most N-terminal hyaluronate binding motif of CD44 ablates both hyaluronate and chondroitin sulfate binding, suggesting that glycosaminoglycans are bound through a common motif, and that only one of the hyaluronate binding motifs is responsible for the majority of glycosaminoglycan binding by CD44 on the cell surface. Taken together, these observations indicate that alternative splicing regulates the ligand binding specificity of CD44 and suggest that structural changes in the CD44 protein have a profound effect on the range of ligands to which this molecule can bind with potentially wide-ranging functional consequences. PMID- 9395531 TI - A mutation in the aryl hydrocarbon receptor (AHR) in a cultured mammalian cell line identifies a novel region of AHR that affects DNA binding. AB - Introduction of a retroviral expression vector for the aryl hydrocarbon receptor (AHR) restores CYP1A1 inducibility to a mutant derivative of the Hepa-1 cell line that is defective in induction of CYP1A1 by ligands for the receptor. An AHR protein with normal ligand binding activity is expressed in the mutant but ligand treatment of mutant cell extract fails to induce binding of the AHR. ARNT (aryl hydrocarbon receptor nuclear translocator) dimer to the xenobiotic responsive element (XRE). AHR cDNAs derived from the mutant encode a protein that is unimpaired in ligand-dependent dimerization with ARNT, but the AHR.ARNT dimer so formed is severely impaired in XRE binding activity. The mutant cDNAs contain a C to G mutation at base 648, causing a cysteine to tryptophan alteration at amino acid 216, located between the PER-ARNT-SIM homology region (PAS) A and PAS B repeats. Introduction of the same mutation in the wild-type AHR sequence by site directed mutagenesis similarity impaired XRE binding activity. Substitution with the conservative amino acid, serine, had no effect on XRE binding. The tryptophan mutation, but not the wild-type allele, was detectable in genomic DNA of the mutant. The implication that an amino acid within the PAS region may be involved in DNA binding indicates that the DNA binding behavior of AHR may be more anomalous than previously suspected. PMID- 9395532 TI - High affinity dimerization by Ski involves parallel pairing of a novel bipartite alpha-helical domain. AB - c-Ski protein possesses a C-terminal dimerization domain that was deleted during the generation of v-ski, and has been implicated in the increased potency of c ski in cellular transformation compared with the viral gene. The domain is predicted to consist of an extended alpha-helical segment made up of two motifs: a tandem repeat (TR) consisting of five imperfect repeats of 25 residues each and a leucine zipper (LZ) consisting of six heptad repeats. We have examined the structure and dimerization of TR or LZ individually or the entire TR-LZ domain. Using a quenched chemical cross-linking method, we show that the TR dimerizes with moderate efficiency (Kd = 4 x 10(-6) M), whereas LZ dimerizes poorly (Kd > 2 x 10(-5) M). However, the entire TR-LZ domain dimerizes efficiently (Kd = 2 x 10( 8) M), showing a cooperative effect of the two motifs. CD analyses indicate that all three proteins contain predominantly alpha-helices. Limited proteolysis of the TR-LZ dimer indicates that the two helical motifs are linked by a small loop. Interchain disulfide bond formation indicates that both the LZ and TR helices are oriented in parallel. We propose a model for the dimer interface in the TR region consisting of discontinuous clusters of hydrophobic residues forming "leucine buttons." PMID- 9395533 TI - Regulation of leukotriene A4 hydrolase activity in endothelial cells by phosphorylation. AB - Endothelial cells contain leukotriene (LT) A4 hydrolase (LTA-H) as detected by Northern and Western blotting, but several studies have been unable to detect the activity of this enzyme. Since LTA-H could play a key role in determining what biologically active lipids are generated by activated endothelium during the inflammatory process, we studied possible mechanisms by which this enzyme may be regulated. We find that LTA-H is phosphorylated under basal conditions in human endothelial cells and in this state does not exhibit epoxide hydrolase activity (i.e. conversion of LTA4 to LTB4). LTA-H purified from endothelial cells is efficiently dephosphorylated by incubation with protein phosphatase-1 in the presence of an LTA-H peptide substrate and not at all in the absence of substrate. Under conditions that lead to dephosphorylation, protein phosphatase-1 activates the epoxide hydrolase activity of LTA-H. Using peptide mapping and site directed mutagenesis, we have identified serine 415 as the site of phosphorylation of LTA-H by a kinase found in endothelial cell cytosol. In parallel, we have studied a human lung carcinoma cell line that expresses active LTA-H. Although these cells have cytosolic kinases that phosphorylate recombinant LTA-H, they do not target serine 415 and thus do not inhibit LTA-H activity. We believe that LTA-H is regulated in intact cells by a kinase/phosphatase cycle and further that the kinase in endothelial cells specifically recognizes and phosphorylates a regulatory site in the LTA-H. PMID- 9395534 TI - Identification of cysteine pairs within the amino-terminal 5% of apolipoprotein B essential for hepatic lipoprotein assembly and secretion. AB - There is growing evidence that the amino-terminal globular domain of apolipoprotein B (apoB) is essential for lipoprotein particle formation in the hepatic endoplasmic reticulum. To identify the structural requirements for its function in lipoprotein assembly, cysteine (Cys) pairs required to form the seven disulfide bonds within the amino-terminal 21% of apoB were replaced in groups or individually by serine. Substitution of Cys pairs required for formation of disulfide bonds 1-3 or 4-7 (numbered from amino to carboxyl terminus) completely blocked the secretion of apoB28 in transfected HepG2 cells. To identify the specific disulfide bonds required for secretion, Cys pairs were mutated individually. Substitution of Cys pairs required for disulfide bonds 1, 3, 5, 6, or 7 had little or no impact on apoB28 secretion or buoyant density. In contrast, individual substitution of Cys pair 2 (amino acid residues 51 and 70) or 4 (218 and 234) severely inhibited apoB28 secretion and its capacity to undergo intracellular assembly with lipid. The same assembly and secretion defects were observed when these mutations were expressed as part of apoB50. These studies provide direct evidence that the ability of the internal lipophilic regions of apoB to engage in the recruitment and sequestration of lipid during translation is critically dependent upon a structural configuration contained within or affected by the amino-terminal 5% of the protein. PMID- 9395535 TI - Yrb2p is a nuclear protein that interacts with Prp20p, a yeast Rcc1 homologue. AB - A conserved family of Ran binding proteins (RBPs) has been defined by their ability to bind to the Ran GTPase and the presence of a common region of approximately 100 amino acids (the Ran binding domain). The yeast Saccharomyces cerevisiae genome predicts only three proteins with canonical Ran binding domains. Mutation of one of these, YRB1, results in defects in transport of macromolecules across the nuclear envelope (Schlenstedt, G., Wong, D. H., Koepp, D. M., and Silver, P. A. (1995) EMBO J. 14, 5367-5378). The second one, encoded by YRB2, is a 327-amino acid protein with a Ran binding domain at its C terminus and an internal cluster of FXFG and FG repeats conserved in nucleoporins. Yrb2p is located inside the nucleus, and this localization relies on the N terminus. Results of both genetic and biochemical analyses show interactions of Yrb2p with the Ran nucleotide exchanger Prp20p/Rcc1. Yrb2p binding to Gsp1p (yeast Ran) as well as to a novel 150-kDa GTP-binding protein is also detected. The Ran binding domain of Yrb2p is essential for function and for its association with Prp20p and the GTP-binding proteins. Taken together, we suggest that Yrb2p may play a role in the Ran GTPase cycle distinct from nuclear transport. PMID- 9395536 TI - The tryptase, mouse mast cell protease 7, exhibits anticoagulant activity in vivo and in vitro due to its ability to degrade fibrinogen in the presence of the diverse array of protease inhibitors in plasma. AB - Mouse mast cell protease (mMCP) 7 is a tryptase of unknown function expressed by a subpopulation of mast cells that reside in numerous connective tissue sites. Because enzymatically active mMCP-7 is selectively released into the plasma of V3 mastocytosis mice undergoing passive systemic anaphylaxis, we used this in vivo model system to identify a physiologic substrate of the tryptase. Plasma samples taken from V3 mastocytosis mice that had been sensitized with immunoglobulin (Ig) E and challenged with antigen were found to contain substantial amounts of four 34-55-kDa peptides, all of which were derived from fibrinogen. To confirm the substrate specificity of mMCP-7, a pseudozymogen form of the recombinant tryptase was generated that could be activated after its purification. The resulting recombinant mMCP-7 exhibited potent anticoagulant activity in the presence of normal plasma and selectively cleaved the alpha-chain of fibrinogen to fragments of similar size as that seen in the plasma of the IgE/antigen-treated V3 mastocytosis mouse. Subsequent analysis of a tryptase-specific, phage display peptide library revealed that recombinant mMCP-7 preferentially cleaves an amino acid sequence that is nearly identical to that in the middle of the alpha-chain of rat fibrinogen. Because fibrinogen is a physiologic substrate of mMCP-7, this tryptase can regulate clot formation and fibrinogen/integrin-dependent cellular responses during mast cell-mediated inflammatory reactions. PMID- 9395537 TI - Dephosphorylation of the cadherin-associated p100/p120 proteins in response to activation of protein kinase C in epithelial cells. AB - Protein kinase C signaling pathways have been implicated in the disruption of intercellular junctions, but mechanisms are not clear. p100 and p120 are members of the Armadillo family of proteins and are localized to cellular adherens junctions. In strain I Madin-Darby canine kidney cells, protein kinase C activation leads to disruption of tight junctions and an increase in permeability of cell monolayers. We show that this permeability increase is accompanied by dephosphorylation of p100/p120 on serine and threonine residues. The dephosphorylation of these proteins can also be induced by the kinase inhibitors staurosporine, KT5926, and Go 6976. Treatment of cells with phosphatase inhibitors induced hyperphosphorylation of p100 and p120. Thus, p100 and p120 participate in a regulatable cycle of serine/threonine phosphorylation and dephosphorylation. Protein kinase C must act, directly or indirectly, by perturbing this phosphorylation cycle, by inhibition of a p100/p120 kinase and/or activation of a phosphatase. These data clearly show that p100 and p120 are targets of a novel protein kinase C signaling pathway. Dephosphorylation of these proteins precedes the permeability increase across epithelial cell monolayers seen in response to phorbol esters, raising the possibility that this pathway may play a role in the modulation of intercellular junctions. PMID- 9395538 TI - Drosophila factor 2, an RNA polymerase II transcript release factor, has DNA dependent ATPase activity. AB - Drosophila factor 2 has been identified as a component of negative transcription elongation factor (N-TEF) that causes the release of RNA polymerase II transcripts in an ATP-dependent manner (Xie, Z. and Price D. H. (1996) J. Biol. Chem. 271, 11043-11046). We show here that the transcript release activity of factor 2 requires ATP or dATP and that adenosine 5'-O-(thiotriphosphate) (ATPgammaS), adenosine 5'-(beta,gamma-imino)triphosphate (AMP-PNP), or other NTPs do not support the activity. Factor 2 demonstrated a strong DNA-dependent ATPase activity that correlated with its transcript release activity. At 20 microg/ml DNA, the ATPase activity of factor 2 had an apparent Km(ATP) of 28 microM and an estimated Kcat of 140 min-1. Factor 2 caused the release of nascent transcripts associated with elongation complexes generated by RNA polymerase II on a dC tailed template. Therefore, no other protein cofactors are required for the transcript release activity of factor 2. Using the dC-tailed template assay, it was found that renaturation of the template was required for factor 2 function. PMID- 9395539 TI - The VRG4 gene is required for GDP-mannose transport into the lumen of the Golgi in the yeast, Saccharomyces cerevisiae. AB - In the yeast Saccharomyces cerevisiae, glycoproteins and sphingolipids are modified in the Golgi by the addition of mannose residues. The critical mannosyl donor for these reactions is the nucleotide sugar, GDP-mannose, whose transport into the Golgi from the cytoplasm is required for mannosylation. This transport reaction has been well characterized, but the nucleotide sugar transporter has yet to be identified in yeast. VRG4 is an essential gene whose product is required for a number of Golgi-specific functions, including glycosylation and the organization of the endomembrane system. Here, data are presented that demonstrate that the primary role of Vrg4p is in the transport of GDP-mannose into the Golgi. The vrg4 mutation causes a general impairment in mannosylation, affecting N-linked and O-linked glycoprotein modifications as well as the mannosylation of sphingolipids. By using an in vitro assay, vrg4 mutants were shown to be specifically defective in the transport of GDP-mannose into Golgi vesicles. The Vrg4 protein localizes to the Golgi complex in a pattern that suggests a wide distribution throughout the Golgi. Vrg4p displays homology to other putative nucleotide sugar transporters, suggesting that the VRG4 gene encodes a Golgi GDP-mannose transporter. As Vrg4p is essential, these results suggest that a complete lack of mannosylation of glycoproteins in the Golgi leads to inviability. Alternatively, the essential function of Vrg4p in yeast involves its effect on sphingolipids, which would imply a critical role for mannosylinositol phosphorylceramides or mannosyl diphosphoinositol ceramides on growth and viability. PMID- 9395540 TI - S100beta inhibits alpha1-adrenergic induction of the hypertrophic phenotype in cardiac myocytes. AB - In an experimental rat model of myocardial infarction, surviving cardiac myocytes undergo hypertrophy in response to trophic effectors. This response involves gene reprogramming manifested by the re-expression of fetal genes, such as the previously reported isoform switch from adult alpha- to embryonic beta-myosin heavy chain. We now report the transient re-expression of a second fetal gene, skeletal alpha-actin in rat myocardium at 7 days post-infarction, and its subsequent down-regulation coincident with the delayed induction of S100beta, a protein normally expressed in brain. In cultured neonatal rat cardiac myocytes, co-transfection with an S100beta-expression vector inhibits a pathway associated with hypertrophy, namely, alpha1-adrenergic induction of beta-myosin heavy chain and skeletal alpha-actin promoters mediated by beta-protein kinase C. The induction of beta-myosin heavy chain by hypoxia was similarly blocked by forced expression of S100beta. Our results suggest that S100beta may be an intrinsic negative regulator of the hypertrophic response of surviving cardiac myocytes post-infarction. Such negative regulators may be important in limiting the adverse consequences of unchecked hypertrophy leading to ventricular remodeling and dysfunction. PMID- 9395541 TI - Activation by cyclic GMP binding causes an apparent conformational change in cGMP dependent protein kinase. AB - Cyclic nucleotide binding activates cyclic nucleotide-dependent protein kinases, but the molecular mechanism is unknown. In the present studies, cGMP binding to type Ialpha or type Ibeta cGMP-dependent protein kinase (PKG) caused (i) a large electronegative charge shift of each enzyme on ion exchange chromatography, (ii) an increase in the Stokes radius (>3 A) of each enzyme, and (iii) a decreased mobility of type Ibeta PKG on native gel electrophoresis. These physical changes were not detected in the monomeric form of type Ibeta PKG upon activation by cGMP. However, the results of partial proteolysis of type Ialpha PKG revealed some degree of cGMP-induced conformational change within the PKG-monomer, since cGMP binding protects the PKG-monomer against chymotryptic cleavage. The altered sensitivity to proteolysis occurs at Met-200, which is located between the B and C alpha-helices in the high affinity site (site A), and implies that the cGMP induced structural perturbations in this region may participate in activation of dimeric PKG. The cGMP-induced conformational effects observed using the physical separation methods are likely to reflect altered interactions within the dimeric PKG that are caused by structural alterations within the subunits. PMID- 9395542 TI - Progressive cyclic nucleotide-induced conformational changes in the cGMP dependent protein kinase studied by small angle X-ray scattering in solution. AB - Small angle scattering data from bovine lung type Ialpha cGMP-dependent protein kinase (PKG) in the absence of cGMP show the protein to have a highly asymmetric structure with a radius of gyration (Rg) of 45 A and a maximum linear dimension (dmax) of 165 A. The addition of cGMP induces a marked conformational change in PKG. The Rg and dmax increase 25-30%, and the protein's mass moves further away from the center of mass; this results in an even more asymmetric structure. Fourier transform infrared spectroscopy data suggest that the conformational change induced by cGMP binding is primarily due to a topographical movement of the structural domains of PKG rather than to secondary structural changes within one or more of the individual domains. Each monomer of the dimeric PKG contains one high and one low affinity cGMP-binding site. A prominent increase in the asymmetry of PKG occurs with binding to high affinity cGMP-binding sites alone, but the full domain movements require the binding to both sets of sites. These conformational changes occurring in PKG with the progressive binding of cGMP to both sets of cGMP-binding sites correlate with past data, which have indicated that cGMP binding to both sets of sites is required for the full activation of the enzyme. These results provide the first quantitative measurement of the overall PKG structure, as well as an assessment of the structural events that accompany the activation of a protein kinase upon binding a small molecular weight ligand. PMID- 9395543 TI - Ethnicity and health: the challenges ahead. PMID- 9395544 TI - Coronary heart disease risk factors in black and white patients with non-insulin dependent diabetes mellitus. AB - OBJECTIVE: To determine possible racial differences in risk factors for coronary heart disease (CHD) in black and white patients with noninsulin-dependent diabetes mellitus (NIDDM). METHODS: Study of risk factors for coronary heart disease among 308 subjects who met the WHO criteria for NIDDM. RESULTS: Both black and white patients were found to have a high prevalence of hypertension, obesity, low high density lipoprotein (HDL) cholesterol, low leisure-time physical activity levels, and an atherogenic dietary profile. Black males were more likely to have hypertension, reported a greater intake of dietary cholesterol, and had lower triglycerides, higher HDL cholesterol levels, a lower CHOL/HDL ratio, and a lower waist to hip ratio (WHR) than white males. Black females had higher mean arterial and diastolic blood pressures, had lower triglycerides, higher HDL cholesterol, a lower CHOL/HDL ratio, a higher subscapular/triceps ratio and lower reported leisure-time energy expenditure compared to white females. There were no racial differences found for obesity level. CONCLUSION: Our results indicate that racial differences in CHD risk factors exist among black and white patients with NIDDM. The complex genetic, sociocultural and environmental interactions involving CHD risk factors that contribute to the development of CHD may eventually provide clues to the etiology of the disease. PMID- 9395545 TI - Racial differences in the incidence of end-stage renal disease. AB - OBJECTIVE: To examine trends in the incidence of treated end-stage renal disease (ESRD) and variations between blacks and whites. DESIGN: Retrospective record reviews of all new patients > or = 15 years starting chronic dialysis during 1980 1988 at the Piedmont Dialysis Center, Forsyth County, North Carolina. RESULTS: The cumulative nine-year incidence rate for hypertensive ESRD was 570 per million, and for diabetic ESRD 497 per million. Among men, hypertensive ESRD accounted for the largest proportion of cases (39.2% and 28.4%, blacks and whites respectively), while diabetic ESRD contributed 33.9% of black female cases and 24.4% of white female cases. Compared to whites, blacks were at significantly increased risk, with an adjusted risk odds ratio (OR) of 4.4 (95% confidence interval (CI) 3.5-6.0) for all causes combined, 6.0 (CI 3.9-9.0) for hypertensive renal disease, 6.0 (CI 3.8-9.3) for renal disease due to insulin-dependent diabetes mellitus, and 12.2 (CI 6.9-21.7) due to non-insulin dependent diabetes mellitus (NIDDM). The greatest risk among blacks was seen in the 55-64 year age group, with ORs of 9.1 for all causes combined and 30.6 for hypertensive renal disease. The OR for renal disease due to NIDDM for black versus white women was 20.0 (CI 9.5-41.7). Compared to 1980, 1981, 1982 and 1983, increased incidence rates were seen in each year after 1984. CONCLUSION: These findings show even greater excess risk of ESRD among blacks than previously reported. The majority of the excess risk is seen for ESRD due to hypertension and diabetes, especially NIDDM. The reasons for the increased risk among blacks, and for the increasing incidence rates of ESRD are not known. PMID- 9395546 TI - Relationship of hypertension to socioeconomic status in a west African population. AB - OBJECTIVES: To determine whether hypertension rates were positively related to socioeconomic status (SES) in males in urban northern Nigerian civil servants in order to confirm this relationship previously observed in a southern Nigerian civil servant population which differed in tribal origin, religious practices and diet. METHODS: Civil servants were recruited from the Sokoto State ministries, Sokoto, Nigeria. Professionals and administrators were designated as higher SES, and clerks and laborers as lower SES. In addition to blood pressure, the height and weight of individuals, as well as their urinary sodium- and potassium creatinine, were also measured. RESULTS: The age-adjusted occurrence of hypertension (systolic pressure > or = 140 mmHg or diastolic pressure > or = 90 mmHg or current use of hypertension medication) was similar in male higher (n = 155) and lower (n = 255) SES groups aged 25-54, 19.3% and 19.8%, respectively. However, the age-adjusted rate of definite hypertension (systolic pressure > or = 160 mmHg or diastolic pressure > or = 95 mmHg or current use of hypertension medication) was considerably higher in the higher SES than in the lower SES men, 11.2% versus 3.6%. Age-adjusted body mass index (BMI, kg/m2) was higher among the higher than in the lower SES group, 21.4 versus 20.4. Over-night sodium excretion did not differ. Among female civil servants (n = 73) aged 20-44, there were few of higher SES (n = 19) precluding SES-specific analyses. Total and definite hypertension rates among women were 17.2% and 5.5%, respectively. Mean BMI was 22.2. In logistic regression, definite hypertensive status was related to age group, BMI tertile, sodium excretion and SES in men and to sodium excretion in women. CONCLUSION: Even in this very lean population, the higher risk for hypertension in males of higher SES was confirmed. This was explained, in part, by higher BMI. PMID- 9395547 TI - Ethnic differences in public health awareness, health perceptions and physical exercise: implications for heart disease prevention. AB - OBJECTIVES: To study whether the wide differences in heart disease incidence amongst ethnic groups in the UK, with the higher mortality and morbidity in peoples of south Asian descent, may be attributed to differences in public health awareness and life-style. DESIGN: Questionnaire-based survey of women from different ethnic groups attending an antenatal clinic in a city centre district general hospital. RESULTS: We surveyed 232 housewives from 3 different ethnic groups: 84 white (mean age 24.3 years +/- standard deviation (SD) 5.84), 76 Afro Caribbean (mean age 24.7 years +/- SD 4.46) and 72 Asians (defined as people of south Asian or Indian subcontinent descent; mean age 25.7 years +/- SD 5.5). The proportions of smokers amongst the whites, blacks and Asians were 38.1%, 27.6% and 2.8% respectively. Proportions consuming alcohol regularly were 31.0%, 10.5% and 4.2% respectively. A higher proportion of blacks reported a change in dietary fibre intake, whilst a higher proportion of whites reported a change in dietary fat intake and sugar intake as a result of public health campaigns, publicity or advertising. There was a significantly lower proportion of reported regular exercise activity amongst the Asian women and their husbands or partners. CONCLUSION: This study demonstrates that Asian families were the least likely to take regular exercise, and had a lower awareness of cholesterol or dietary content (fibre, sugar, salt) despite public health campaigns and publicity. They were however the least likely to smoke cigarettes. These ethnic differences may in part explain the higher prevalence of coronary heart disease amongst the Asian population in the UK. This ethnic group should be targeted for intense public health intervention, education and other preventative measures to reduce the risks of heart disease. PMID- 9395548 TI - Suicide patterns and trends in people of Indian subcontinent and Caribbean origin in England and Wales. AB - OBJECTIVES: To examine suicide rates and trends in people of Indian subcontinent, east African and Caribbean origin using the latest mortality data available for England and Wales. To compare suicide rates in these groups with the baseline and target rates for suicide in the Health of the Nation strategy. METHODS: Suicide data for England and Wales for 1988-1992, classified by the country of birth of the deceased, and population denominators from the 1991 Census were used for the analysis. Standardised mortality ratios (SMRs) for ages 15-64 and age-specific ratios were computed, using the age-sex specific rates for England and Wales as the standard. Trends over the preceding decade and suicide by burning were also analysed. Directly age-standardised suicide rates were derived to facilitate comparison with Health of the Nation baseline and target rates. RESULTS: Suicide ratios were significantly low (SMRs 32, 52 and 55 respectively) in Bangladeshi, Sri Lankan and Pakistani born men at all ages, but raised in young Indian and east African men. Ratios were significantly high in Indian and east African women (143 and 154), with a 2-3 fold excess at ages 15-34 years. Ratios were low in Pakistani and Bangladeshi women overall, but elevated at 15-24 years. For the Caribbean-born, ratios were low overall but raised at ages 25-34. 20% of Asian female suicides were by burning. Indians are a high risk group in terms of the Health of the Nation suicide targets. Suicide trends in the minority ethnic groups reflect national trends. CONCLUSIONS: This study confirms previous findings of high suicide rates in young Asian women. A new finding is the raised suicide rate in young Caribbeans. High suicide risks among young people from some ethnic minority communities are significant in the context of both the Health of the Nation strategy and recent governmental concern about the need to tackle health variations in the UK. Such deaths are indicative of larger numbers of young ethnic minority adults at risk of mental distress and self harm. PMID- 9395549 TI - Race, socioeconomic status and survival in three female cancers. AB - OBJECTIVES: Although many studies have reported that socioeconomic status (SES) and race affect cancer survival, researchers have not established whether SES and race affect survival independently. The research reported here addresses this question with special attention to cancers affecting large numbers of women in the US. METHODS: The authors analyzed data on survival among patients in the Centralized Cancer Patient Data System (CCPDS) with cancers of the breast (n = 6896), cervix (n = 2209) and uterine corpus (n = 1492). RESULTS: According to Cox proportional hazards models, race predicted survival in all three cancers, while socioeconomic status predicted survival for cancers of the breast and uterine corpus. Interaction effects between race and SES were generally not statistically significant. This study includes larger numbers of observations within specific forms of cancer and covers a broader patient population than most previous investigations. These features promote detectability of SES effects, comparability among disease sites, and generalizability to cancer patients throughout the US. CONCLUSIONS: Findings imply that SES and race affect cancer mortality risk independently of each other, and that the impact of SES and race may vary by malignancy. Survival disadvantages due to race-which may be more pronounced among women than men-should remain a continuing concern. PMID- 9395550 TI - Gender and ethnic differences in survival in a cohort of HIV positive clients. AB - OBJECTIVES: The purpose of this study was to examine gender and ethnic differences in survival of persons receiving treatment for HIV infection to determine if differences existed, and if they did, to assess the possibility of explaining these differences by examining other factors, such as age, disease severity when beginning treatment, alcohol, illicit drugs, tobacco, educational level, living arrangements, antiretroviral treatment, PCP prophylaxis, sexually transmitted diseases, mode of transmission and opportunistic infections. DESIGN: A retrospective cohort study of all clients receiving treatment at an HIV only clinic from its opening in early 1988 until the end of May 1993. Statistical methods used to examine the data included incidence density ratios, Kaplan-Meier survival curves, Breslow (generalized Wilcoxon) tests of equality of survival curves and Cox proportional hazards models both with and without time dependent covariates. RESULTS: In the cohort (37% African American, 7% Hispanic American and 25% female), 220 deaths occurred during 1223 person years of follow-up. Compared to European American males, the following incidence density ratios were observed: European American females: 0.50, Hispanic American females: 0.70, Hispanic American males: 0.96, African American females: 1.28 and African American males: 2.38. The differences were noted above for gender/ethnicity groups were significant at the p < 0.0001 level. After adjusting for disease stage (as measured by laboratory testing of CD4 positive T-lymphocytes), educational level, and age, no differences in survival by gender or ethnicity remained. Disease stage and educational level had the greatest prognostic significance. CONCLUSIONS: European Americans entered treatment at a much earlier disease stage (as measured by CD4 positive T-lymphocyte counts) and had higher educational levels (a surrogate for socioeconomic status) than African Americans. These factors may explain the longer survival in European Americans as compared to African Americans in this cohort. PMID- 9395551 TI - Race/ethnicity misclassification of persons reported with AIDS. The AIDS Mortality Project Group and The Supplement to HIV/AIDS Surveillance Project Group. AB - OBJECTIVES: To examine differences in race/ethnicity classifications of persons with AIDS among three reporting sources and to estimate the effect of these differences on calculated AIDS rates. METHODS: We reviewed case reports from the national AIDS surveillance database, interview (self-reported) data from 11 state/local health departments, and death certificate information from 16 state/local health departments for agreement in race/ethnicity coding among persons reported with AIDS. RESULTS: Race/ethnicity coding inconsistencies with AIDS case reports were greatest for persons identified as American Indians/Alaska natives on death certificates (46% [47/102] disagreement) and by self-report (57% 8/14 disagreement). Agreement with AIDS case reports was highest either for persons identified as white from death certificates (4% [1314/31,070] disagreement) and white from self-reports (2% [26/1068] disagreement) or black from death certificates (3% [440/13,592] disagreement) and black from self reports (3% [21/736] disagreement). For other racial/ethnic groups, disagreement with AIDS case reports was intermediate; for Asians/Pacific Islanders, 12% [45/377] disagreement with death certificates and 33% 4/12 disagreement with self reports; and for Hispanics, 14% [1151/8527] disagreement with death certificates and 24% [59/249] disagreement with self-reports. CONCLUSION: For certain racial/ethnic groups, classification by race/ethnicity can differ substantially by surveillance data source. Because allocation of public health resources may be determined by estimates of disease impact on different population groups, periodic evaluations of the accuracy of race and ethnicity reporting are needed to assure appropriate distribution of these resources. PMID- 9395552 TI - Use of the terms 'race', 'ethnicity', and 'national origins': a review of articles in the American Journal of Public Health, 1980-1989. AB - OBJECTIVES: To assess the use of the categories of race, ethnicity, and national origin in recent public health research. METHODS: We reviewed all research articles on human populations published in the American Journal of Public Health from January 1980 through December 1989. Articles were classified by (1) mention of the categories, (2) use of the categories, (3) presence of explicit definitions, and (4) definitional criteria. RESULTS: Specific categories (e.g. 'black', 'Chinese', 'Hispanic') or generic categories (e.g. 'race', 'ethnicity', 'national origin') were mentioned in 461 (50.4%) of 914 articles on human populations. In most studies (65.1%), single categories (e.g. race or ethnicity) were considered; in 1.3% of studies, two terms (e.g. both race and ethnicity) were examined independently; in 1.3%, categories were used interchangeably; in 5.6% of the studies, combined categories (e.g. race-ethnicity) were used; and in 27.5% of the studies, specific population groups were named without reference to a generic category. Explicit definitions of categories were present in only 8.4% of the articles in which the categories were considered. Absence of explicit definitions and use of combined and interchangeable categories suggest a lack of clarity and conceptual consistency in research on race, ethnicity, and national origin-related topics. CONCLUSION: To improve our assessment of differences in health status among racial, ethnic, and national origin groups, research involving these categories should assess their validity and should define concepts clearly, explicitly, and consistently. Such research would minimize misclassification, improve the interpretation of findings, facilitate comparison among studies, and enhance the understanding of causes underlying differences in health status among populations of different racial, ethnic, and national origins. PMID- 9395553 TI - Utilization of health care services by immigrants and other ethnic/cultural groups in Ontario. AB - OBJECTIVES: This study assesses the accessibility of health care services by immigrants and other ethnic/cultural groups in Ontario, using the 1990 Ontario Health Survey. METHODS: The population sample of 38,519 adults aged 16-64 is weighted to represent the entire non-institutionalized population of the province. Outcome measures were whether the study participants visited a general practitioner's office, a specialist's office, or a hospital's emergency department during the past 12 months. RESULTS: The results showed that while the percentages of participants who ever visited a general practitioner's office during the past 12 months were slightly higher in immigrants and other ethnic/cultural groups, the rates of visits to the specialist's office were quite similar, and the rates of hospital emergency department's visits were often lower (except for aboriginals), than for Canadians. These differences in the utilization of health services across different immigrant and ethnic/cultural groups remained unchanged after controlling for health status (as measured by self-reported health problems) and age differences. However, because the sample sizes in some immigrant and ethnic/cultural groups were small, many of the differences were not statistically significant. CONCLUSIONS: We conclude that while immigrants and other ethnic/cultural groups in Ontario usually had equal access to regular services (e.g., visits to general practitioner's office), they often had lower utilization of hospital emergency departments. However, general purpose surveys have limited utility in assessing reasons of health care utilization amongst different ethnic/cultural groups. PMID- 9395554 TI - Hypertension in African Americans: basic science initiatives of the National Heart, Lung and Blood Institute. PMID- 9395555 TI - Hereditary intermediate phenotypes in African American hypertension. AB - OBJECTIVE: Essential hypertension is a heterogeneous and multifactorial disorder and is at least twice as frequent among African Americans as in the general population. Inheritance of high blood pressure is complex, with the gene(s) responsible for hypertension still remaining elusive. A useful strategy for investigating the heritability of hypertension is to evaluate 'intermediate phenotypes'--simple Mendelian or monogenic traits that are associated with hypertension. These intermediate steps may identify potential pathophysiological factors that antedate the development of high blood pressure and suggest candidate genes. We are attempting to identify and characterize several such intermediate phenotypes, in particular as these might apply to hypertension in African Americans. METHODS: We studied several physiological and biochemical candidate intermediate phenotypes in untreated black and white patients with essential hypertension and in their normotensive counterparts stratified by genetic risk of hypertension. RESULTS AND CONCLUSIONS: Promising intermediate phenotypes, which may be useful for studies in African American families, include baroreceptor sensitivity to low and high pressure stimuli, cold pressor test responses, and biochemical markers such as plasma chromogranin A, dopamine-beta hydroxylase and urinary kallikrein excretion. Identification of genes involved in complex traits such as hypertension may be facilitated by the intermediate phenotype approach, combined with recent advances in quantitative genetics and linkage mapping. Further studies are needed to pinpoint the nature of genes in African American hypertension. PMID- 9395556 TI - New methods for maintaining human renal epithelial cells and analyzing their ion transport functions: potential analysis of genetic disease. AB - OBJECTIVES: New methods are available to immortalize parenchymal cells from exocrine glands and kidney with retention of differentiation. Adaptation of this technology to small, single-donor biopsy material or surgical specimens could provide genetically homogeneous cells for functional analyses and correlation with genetic background and underlying biochemistry. To develop a methodology useful for renal sodium metabolism, epithelial cell line generation was tested in a hypertensive rat model with features similar to salt-sensitive hypertension in humans. This form of hypertension has a large genetic component and is prevalent in African Americans. DESIGN: Protocols were designed to immortalize primary cultures of microdissected proximal tubule epithelial cells from spontaneously hypertensive (SHR) and control, normotensive Wistar-Kyoto (WKY) rats. Immortalization was based on a replication-defective retrovirus coding for SV40 large T-antigen as positive cell cycle regulator. Transport competent cells that grow on porous filters to form confluent monolayers were selected. RESULTS: Several proximal tubule cell lines have been developed from SHR and WKY rats. The cells retain important differentiated features, such as epithelial polarity, low monolayer conductance, and sodium-succinate cotransport. They are suitable for analyses of electrolyte transport by electrophysiology or imaging of intracellular fluorescent indicator dyes, such as sodium-binding benzofuran isophthalate. CONCLUSION: Feasibility of generating epithelial cell lines from defined renal segments was demonstrated. The cells retain important transport function so that analyses of sodium metabolism and the influence of genetic background on it are possible. The methodology is applicable to human specimens. PMID- 9395557 TI - Intracellular cations and hypertension in blacks. AB - Considerable attention has been focused in recent years on the role of intracellular ions in the pathophysiology of hypertension in African Americans. Following the identification of marked differences in red cell sodium content and sodium-lithium counter-transport between blacks and whites, the hypothesis emerged that cation metabolism at the cellular level might account for part of the ethnic difference in susceptibility to hypertension. Unfortunately, findings in the red cell have not significantly increased our understanding of the physiologic pathways and may prove to be anomalous. Interest has recently shifted to calcium metabolism, because of its greater physiologic relevance, and a new series of questions are being defined. With the development of fluoroprobes to measure sodium and calcium simultaneously, wider application of pharmacologic agonists and the use of single cell techniques have provided a new direction for this field. Whether the ethnic contrasts observed for sodium in the red cell can be reproduced in platelets remains to be seen. It may be that the earlier red cell differences were epiphenomena, unrelated to the control of blood pressure, and that the more closely one approaches basic physiologic mechanisms, the greater the cross-ethnic group similarity. Rather than drawing attention to unusual phenotypic characteristics, the study of hypertension in blacks may be more instructive for the insight it provides into the basic physiology of this disorder, common to all ethnic groups. PMID- 9395558 TI - The kidney in the hypertensive black. AB - Evidence suggests that the incidence of end-stage renal disease due to essential hypertension is five to six times more frequent in black than in white patients. The reason for this greater susceptibility is not clear. Several possibilities have been proposed, including socioeconomic factors, compliance with therapy, renal hemodynamic differences and anatomic differences. In this review, we propose that the greater propensity of black hypertensives to develop renal failure as a consequence of hypertension may be due to abnormal hemodynamic adaptation of the renal circulation to a rise in blood pressure caused by high dietary sodium intake. This would make the renal circulation of hypertensive blacks more susceptible to injury. PMID- 9395559 TI - Excess admixture proportion of extended major histocompatability complex haplotypes of Caucasian origin among rheumatoid arthritis associated haplotypes in African Americans and Afro-Caribbeans. AB - Several extended major histocompatability complex (MHC) haplotypes are associated with susceptibility to autoimmune disease in Caucasian populations. It is known that African Americans and Afro-Caribbeans are ethnic groups descended from west, central and southern black African populations which are admixed with Caucasians. To examine the possible association of some marker of Caucasian MHC genes and susceptibility to rheumatoid arthritis (RA) in African Americans, we studied extended MHC haplotypes (HLA-B, complement and DR) in a sample of 18 African American and Afro-Caribbean probands with RA, their first degree relatives and in 15 non-RA families. We defined 36 disease-associated RA haplotypes among the probands and 96 normal haplotypes in normal individuals. To obtain the most conservative estimate, we excluded recognized Caucasian, DR4-bearing, extended MHC haplotypes from the analysis. Admixture proportions for non-HLA-DR4 extended MHC haplotypes of known Caucasian origin among RA-associated and normal haplotypes were computed (0.40 versus 0.163 respectively). When we compared the difference in proportions between RA and normal haplotypes, the proportion of extended MHC haplotypes of known Caucasian origin was significantly increased among RA-associated haplotypes (Z = 3.16, p (one sided) < 0.001, p (adjusted) < 0.008). Our results suggest that racial admixture with Caucasian MHC genes may augment RA susceptibility and thus may be one mechanism to explain the higher prevalence of RA in African Americans and Afro-Caribbeans than in black African populations. PMID- 9395560 TI - Ethnicity and use of obstetrical analgesia: do Pakistani women receive inadequate pain relief in labour? AB - OBJECTIVE: To study whether the use of analgesic treatment in labour is influenced by ethnicity. DESIGN: A cross-sectional study of hospital patients. Setting; the two municipal hospitals, Ulleval and Aker, in Oslo, Norway. Subjects; a total of 137 obstetrical patients, 67 Pakistani women and 70 Norwegian women. Main outcome measure; use of analgesics in labour. RESULTS: 30% of the Pakistani and 9% of the Norwegian women received no analgesia in labour. Pethidine injection was the preferred analgesic administered to Pakistani women. Women of Pakistani origin received epidural infusion or nitrous oxide and oxygen gas less frequently than Norwegian women. They also received fewer combinations of other analgesic methods. When adjusted for the mothers' age, parity and duration of delivery, Pakistani origin was the only significant predictor for receiving no analgesia in labour. CONCLUSION: Women of Pakistani origin were more than three times as likely not to receive analgesia in labour as Norwegian women. The health services offered to Pakistani women in labour were different from those offered to Norwegian women. These results indicate that women of Pakistani origin may be offered insufficient obstetrical analgesia, or that Norwegian women received unnecessary pain relief in labour. PMID- 9395561 TI - The prevalence of Helicobacter pylori positivity in asymptomatic children of different ethnic backgrounds living in the same country. AB - OBJECTIVE: To measure the prevalence of Helicobacter pylori seropositivity in a population of apparently healthy children of different ethnic backgrounds, matched for age and socioeconomic background, who were born and continue to reside in the same country. DESIGN: The presence of Helicobacter pylori specific IgG antibodies was determined during pre-surgery blood analysis in 883 symptom free children, aged from 1 months to 17 years, who belong to different ethnic populations and were admitted for elective minor surgery. The different groups were matched for age and socioeconomic background. RESULTS: Seventy-two children (8.2%) had a positive titer for Helicobacter pylori. We observed a significant difference in the prevalence of Helicobacter pylori positivity between symptom free Caucasian and non-Caucasian children (p < 0.001). However, no difference could be observed between the non-Caucasian groups (p > 0.8). CONCLUSION: We conclude that significant differences exist in the prevalence of Helicobacter pylori infection between asymptomatic children of different ethnic backgrounds, despite the fact that all investigated subjects were born in Belgium and have been living in this country ever since. Whether this difference is caused by an unknown environmental factor or an, until now, unrecognized genetic predisposition still needs to be evaluated. PMID- 9395562 TI - Sexually transmitted diseases: experience and risk factors among urban, low income, African American and Hispanic youth. AB - OBJECTIVES: The objectives of this study were to assess: (1) ethnic and gender differences in reporting of diagnoses of sexually transmitted diseases (STDs), symptoms related to STDs and sexual behavior and (2) behavioral risk factors for STDs in a probability sample of low income African American and Hispanic youth. METHODS: Data were analyzed from a household probability sample of youth. The study collected data on self reported STDs, symptoms of STDs and sexual behavior. The sample was drawn from households in low income areas of urban Detroit: 1435 African American and Hispanic low income youth age 15-24 living in Detroit were interviewed face to face in 1991. RESULTS: Patterns of sexual activity as well as experience with STDs differ by ethnicity and gender. Within ethnic groups, women report more symptoms of STDs. Risk factors for diagnosed STDs included age, a young age at first intercourse, numbers of sexual partners, oral intercourse and anal intercourse. CONCLUSIONS: The data underscore the importance of development of effective safer sex intervention programs for these youth as well as careful assessment of STD risks in medical clinics serving these youth. PMID- 9395563 TI - Body mass index and cardiovascular risk factors in Pacific Island Polynesians and Europeans in New Zealand. AB - OBJECTIVES: To examine relationships between body mass index (BMI) and cardiovascular risk factors in 279 Europeans and 231 Polynesian Pacific Islanders in New Zealand. METHODS: Participants were recruited from Seventh-Day Adventist church meetings or camps, and were surveyed by self-administered questionnaire. Blood pressure, weight and height were measured. Fasting blood samples were analysed for lipids, glucose and fructosamine. RESULTS: Age-adjusted BMI was higher in Pacific Islanders than in Europeans: 32.8(0.3) versus 25.6(0.3); means(SE); p = 0.0001). In Europeans, BMI was positively associated with systolic and diastolic blood pressures, triglycerides, total cholesterol, LDL cholesterol and fasting blood glucose, and negatively associated with HDL cholesterol. In Pacific Islanders, BMI was associated only with systolic and diastolic blood pressures, and with HDL cholesterol. These associations were stronger in Europeans than in Pacific Islanders. CONCLUSIONS: In this group of Pacific Islanders, the association between BMI and cardiovascular risk factors was weaker than in Europeans. This suggests that either BMI is a poor measure of adiposity in Pacific Islanders, or that adiposity may be less strongly linked to cardiovascular disease in Pacific Islanders. PMID- 9395564 TI - Increased gallbladder-related mortality among Hispanics: does education play a role? AB - OBJECTIVE: Hispanics, particularly Mexican Americans, are known to have a higher incidence of mortalities whose underlying cause is a gallbladder-related disorder. These analyses evaluate the role of educational attainment in the differential mortality experiences of these populations. METHODS: US mortality data for 1989-1991 were examined to determine ethnically-specific death rates using 'any mention' of the disease on the death certificate. RESULTS: Age adjusted multiple cause mortality was found to be higher for all gallbladder related disorders among Hispanics, particularly Mexican Americans. Mortality due to gallbladder cancer, gallstones and 'other gallbladder diseases' were found to be inversely proportional to educational attainment in all ethnic groups. When both age and education were used to adjust mortality, the gallstone and other gallbladder disease mortality among Hispanics was non-significantly higher than white, non-Hispanics. However, mortality due to gallbladder cancer remained significantly higher among Hispanics. CONCLUSION: Gallbladder cancer mortality is elevated in Hispanic populations, especially Mexican Americans, independent of educational attainment. However, increased mortality associated with gallstones or other gallbladder diseases among Hispanics may be partially due to differences in factors associated with educational attainment. Research and public health efforts to address these educational-related factors may improve this mortality pattern among Hispanics. PMID- 9395565 TI - Black-white differences in factors influencing mammography use among employed female health maintenance organization members. AB - OBJECTIVE: This study examined racial differences in knowledge, attitudes and practices related to breast cancer screening of black and white women health maintenance organization members over age 40 who are employed at 75 worksites in Pennsylvania and New Jersey in the US. DESIGN: Data are from telephone interviews of 1677 women (20% black). The interviews queried background factors and concepts from the Health Belief Model. RESULTS: Compared to whites blacks were younger and less likely to be married or to have family history of breast cancer. They were also more likely to underestimate their cancer risk and to fear radiation, and less likely to have a doctor advise them to get mammograms. Black and white women did not differ in terms of self-reported mammography use. The results of multivariate modeling suggest that different set of knowledge and belief variables may explain mammography adherence among black and white women. CONCLUSION: These findings have implications for clinical prevention and for patient and community health education in minority populations. PMID- 9395566 TI - Barriers to follow-up of abnormal screening mammograms among low-income minority women. Cancer Control Center of Harlem. AB - OBJECTIVE: To describe factors related to compliance diagnostic follow-up among minority women of low socioeconomic status with abnormal screening mammograms. METHODS: A retrospective cross-sectional survey using a structured telephone interview. Three cancer screening clinics at an urban inner-city public hospital. All women with abnormal screening mammograms between September 1990 and January 1992 were eligible; women were interviewed in August 1992. Abnormal mammograms were those requiring specific, non-routine clinical follow-up; non-compliance was defined as delayed follow-up (four to six months after the date of the mammogram), or no follow-up at the time of interview (more than 6 months after abnormal). RESULTS: Sixty-two of 442 screened women had abnormal results; the overall rate of non-compliance with follow-up was 50%. Among the 42 (68%) women who agreed to be interviewed, non-compliers were less likely to state that they had been told to receive follow-up than compliers (65% versus 100%; p = 0.008). Non-compliant women were less likely to have suspicious mammography interpretations (p = 0.05), and more likely to report barriers to follow-up, such as cost of lost wages and medical care, system barriers, or fears, than compliant women (61.9% versus 9%, p = 0.01). There were no differences between the two groups for age, education, insurance, source of care, family history, knowledge or attitudes. CONCLUSIONS: These preliminary results suggest that follow-up of low income, minority women with abnormal screening mammograms could be enhanced by improved communication of results. Future studies should extend these findings with larger samples and in other settings and populations. PMID- 9395567 TI - Estimating breast cancer incidence in Hispanic women in Connecticut, 1989-1991. AB - OBJECTIVES: The objective of this study was to estimate incidence rates for breast cancer, the most commonly occurring cancer in women, in the growing Hispanic population of Connecticut. METHODS: The population-based Connecticut Tumor Registry (CTR) routinely obtains only limited information on Hispanic origin and maiden name. In this study, surnames of CTR breast cancer patients diagnosed in 1989-1991 were matched with a list of Spanish surnames. To assess misclassification, both surnames and maiden names (from death certificates) of female Connecticut residents who had died in 1989-1991 from any cause of death at 20 years of age and older were matched with the Spanish-surname list. RESULTS: Age-specific incidence rates (1989-1991) for 'Hispanic' women (with Spanish surname) were lower than those for 'non-Hispanic' white women (with non-Spanish surname) for age 35-39 years and older. Errors in these estimated rates were probably small because among decedents the number with a Spanish surname differed by only 9% from the number with a Spanish maiden name; false positives were almost balanced by false negatives. CONCLUSION: Matching of surnames in the cancer registry with a Spanish surname list provided reasonably accurate estimates of cancer incidence rates in Hispanic women, although individual women were misclassified as 'Hispanic' or 'non-Hispanic'. PMID- 9395568 TI - Appendicitis: higher risk in Mexican Americans? AB - OBJECTIVES: Mexican Americans (MAs), compared to white non-Hispanics (WNHs), have higher rates of biliary disease, noninsulin dependent diabetes, and endstage renal disease but lower rates of lung cancer, hip fractures, and mortality from coronary heart disease. Relatively little research has been done to identify other ethnic differences in disease incidence. We used surgical procedure rates to confirm known ethnic differences and to explore our clinical suspicion that MAs have higher rates of appendectomy than WNHs. METHODS: We used a registry of surgical procedures at two teaching hospitals in South Texas to calculate proportional operation ratios (PORs) for MAs versus WNHs. These two hospitals are the primary source of acute hospital care for the indigent in the area. The POR is arithmetically identical to proportional incidence and mortality ratios. RESULTS: MAs underwent appendectomy proportionally more often than WNHs at both hospitals (POR = 1.41 and 1.75, p < 0.0001). Other significant PORs were consistent with known ethnic disease differences in biliary tract operations, vascular access for chronic hemodialysis, lung cancer, and coronary artery bypass. CONCLUSIONS: These findings support the hypothesis that MAs may undergo appendectomy more often than WNHs and so may be at higher risk of appendicitis. PMID- 9395569 TI - Cultural aspects of African American eating patterns. AB - The high mortality from diet-related diseases among African Americans strongly suggests a need to adopt diets lower in total fat, saturated fat and salt and higher in fiber. However, such changes would be contrary to some traditional African American cultural practices. Focus group interviews were used to explore cultural aspects of eating patterns among low- and middle-income African Americans recruited from an urban community in Pennsylvania. In total, 21 males and 32 females, aged 13-65+ years were recruited using a networking technique. Participants identified eating practices commonly attributed to African Americans and felt that these were largely independent of socioeconomic status. They were uncertain about links between African American eating patterns and African origins but clear about influences of slavery and economic disadvantage. The perception that African American food patterns were characteristically adaptive to external conditions, suggest that, for effective dietary change in African American communities, changes in the food availability will need to precede or take place in parallel with changes recommended to individuals. Cultural attitudes about where and with whom food is eaten emerged as being equivalent in importance to attitudes about specific foods. These findings emphasize the importance of continued efforts to identify ways to increase the relevance of cultural context and meanings in dietary counseling so that health and nutrition interventions are anchored in values as perceived, in this case, by African Americans. PMID- 9395570 TI - Risk-taking behavior among Native American adolescents in Minnesota public schools: comparisons with black and white adolescents. AB - OBJECTIVES: To examine rates of risk-taking behavior among native American adolescents in comparison with blacks and whites, and then to compare our off reservation native American sample to available national statistics on reservation youth. METHODS: A secondary data analysis of a Minnesota public school health survey. Contingency table analyses were performed on a 10% random sample of over 6000 young people focussing on three categories of behavioral risk: antisocial behavior, sexual behavior and substance use. Comparisons were then made to a national convenience sample from reservations and adjacent rural areas. RESULTS: In general, native American adolescents have a significantly higher prevalence of risk behaviors across all indices of antisocial behavior and substance use relative to white and black peers. Native American females presented the most troubling picture. Comparisons to a national convenience sample from reservation lands indicated that native American adolescents in the sample often exceeded national rates of risk behavior. CONCLUSIONS: Residence and attendance at public schools outside reservation lands may make native American adolescents more likely to engage in risky behaviors which endanger their health. PMID- 9395571 TI - Hypertension awareness, treatment and control rates for an Asian population: results from a national survey in Korea. AB - OBJECTIVE: This observational study was performed in order to determine the hypertension awareness, treatment, and control rates for the country of Korea. METHODS: Rates were determined in conjunction with a national blood pressure survey in Korea in 1990. Through cluster sampling, individuals aged > 30 in 190/146,944 districts were selected for study. Among 25,567 eligible individuals, 21,242 had measurement of blood pressure (BP) and answered a standard questionnaire. BP was recorded as the mean of two measurements with a standard mercury manometer. Hypertension was defined either as BP > or = 160/95 mm Hg or on treatment (n = 2628), or as BP > or = 140/90 mm Hg or on treatment (n = 4219). Treatment was defined as any method of BP treatment, including dietary, traditional, or medication. RESULTS: Rates for BP > or = 160/95 mm Hg or on treatment: aware 1057 (40%), treated 696 (27%), controlled 367 (14%). Rates for BP > or = 140/90 mm Hg or on treatment: aware 1069 (25%), treated 696 (16%), controlled 221 (5%). CONCLUSIONS: Hypertension awareness, treatment, and control rates are relatively low in Korea. Blood-pressure control programs, including detection strategies, are needed here and worldwide. PMID- 9395572 TI - Cellular calcium and sodium regulation, salt-sensitivity and essential hypertension in African Americans. AB - The predisposition of African Americans to the salt sensitive form of essential hypertension may result from increased freely exchangeable Ca in intracellular Ca stores and a higher cellular Ca turnover (i.e., enhanced Ca entry into and accelerated Ca extrusion from the cytosol). These alterations entail higher activities of Ca extrusion transport systems, including the Na+/Ca2+ exchanger (NCE), which extrudes Ca in exchange for external Na+, and plasma membrane Ca ATPase (PMCA) that extrudes Ca in exchange for external protons. The higher activity of PMCA, coupled with a higher metabolic activity resulting from a rise in freely exchangeable Ca, increase cellular acid load. Adaptive cellular mechanisms must evolve under these conditions, whereby increased activity of the Na/H exchanger (NHE-1) maintains normal cytosolic pH by enhancing the extrusion of cytosolic protons in exchange for extracellular Na. Cells with increased cellular Ca stores and enhanced Ca turnover may be particularly vulnerable to the factors that inhibit the Na-pump. By inhibiting the Na-pump, these factors diminish the transmembrane Na gradient and consequently inhibit the forward mode of the NCE. Since cells from African Americans show increased Ca turnover, they should retain more Ca upon exposure to Na-pump inhibitors; a heightened sensitivity to Na-pump inhibitors could therefore underlie the propensity of African Americans and other individuals with accelerated cellular Ca turnover rate to the salt sensitive form of essential hypertension. Accelerated cellular Ca turnover in African Americans also explains their better response to Ca antagonists compared with other antihypertensive drugs. PMID- 9395573 TI - Trends in motor vehicle traffic fatalities among Hispanics, non-Hispanic whites and American Indians in New Mexico, 1958-1990. AB - OBJECTIVE: New Mexico has had the highest motor vehicle fatality rate in the nation for many years. Our objective was to examine ethnic differences and trends in motor vehicle fatality rates. DESIGN: Using death certificate data from the New Mexico Bureau of Vital Records and Health Statistics, we compiled age adjusted motor vehicle-related mortality rates from 1958-1990 among the three major ethnic groups in New Mexico--Hispanics, white non-Hispanics and American Indians. RESULTS: Over the 33-year study period, American Indians of both sexes had two to three times higher mortality rates than white non-Hispanics. Hispanic males also had higher motor vehicle death rates than white non-Hispanic males. During the 1970s fatality rates peaked, with age-adjusted death rates of 233/100,000 for American Indian males, 74.7 for Hispanic males and 49.3 for white non-Hispanics for the period 1973-1977. Evaluation of successive 5-year birth cohorts showed highest mortality rates for ages 15-29 years for each ethnic group and both sexes, and a dramatic decline in most ethnic, sex and age-specific rates during the last eight years of the study period. CONCLUSION: Although the recent trends indicate favorable changes in motor vehicle fatality rates, our data highlight the need for ethnic and age-specific interventions to further reduce rates of motor vehicle-related mortality in this state. PMID- 9395574 TI - Racial and ethnic disparities in self-assessed health status: evidence from the National Survey of Families and Households. AB - We examined racial and ethnic disparities in global health assessment and functional limitations of daily activities among whites, blacks and Hispanics, and within the Hispanic origin among Mexicans, Puerto Ricans, Cubans, and 'Others'. Logistic regression were employed to estimate the log odds of reporting 'poor health' and 'having functional limitations' among 12,814 respondents from the 1987-1988 National Survey of Families and Households. Compared with whites, blacks had an increased risk of reporting poor health and functional limitations. Hispanics had even a higher risk of reporting poor health, but did not have an increased risk of reporting functional limitations. Among Hispanics, Mexicans were more likely than whites to report poor health, whereas Puerto Ricans were more likely than whites to experience functional limitations. Both race and ethnicity remain important factors in explaining the disparities in self-assessed health status independent of socioeconomic status (SES). Meanwhile, the way self assessed health status varies with ethnicity is importantly stratified by SES as measured by income and education. These results suggest that future research should analyze the interplay between ethnicity and SES rather than assuming measuring either captures all the important variation. PMID- 9395575 TI - Increased rates of obesity among African Americans confirmed, but the question of why remains unanswered. PMID- 9395576 TI - Obesity in inner-city African Americans. AB - OBJECTIVE: Obesity, a risk factor for chronic diseases, has a high prevalence in African Americans and low-income individuals. However, little is known about perceptions of overweight, attempts to lose weight, and strategies used to lose weight among African Americans in inner cities. DESIGN: A 1990 cross-sectional telephone survey (n = 1445) of north St Louis and central Kansas City, USA. RESULTS: Obesity was common (44%) in this sample of inner-city African Americans. The obese perceived themselves as overweight (70%) and were trying to lose weight (66%). The majority of the obese (68%) were both dieting and exercising to lose weight. Smoking prevalence was not higher among the obese or those trying to lose weight. Many of the obese had received medical advice recently on low-fat diets (51%) and had been advised to lose weight (40%). Factors independently associated with perception, attempts to lose weight and medical advice differed, but included degree of obesity. CONCLUSIONS: These results corroborate US national data that obesity is a public health problem in this population and that obese inner-city African Americans perceive themselves as overweight and are trying to lose weight, especially as degree of obesity increases. It also appears that smoking is not being used as a weight loss strategy and that the obese, as a group, are receiving some medical advice on low-fat diets. This information is critical for designing culturally sensitive weight-control programmes. PMID- 9395577 TI - Differences in weight gain in relation to race, gender, age and education in young adults: the CARDIA Study. Coronary Artery Risk Development in Young Adults. AB - OBJECTIVE: To assess ethnic differences in weight gain in young adults. DESIGN: Five-year weight change was assessed in 4207 young adults initially aged 18-30 years at the CARDIA Study baseline examination (1985-1986). RESULTS: Weight gain was significantly (p < 0.0001) greater in black versus white men (13.2 versus 9.1 lb) and in black versus white women (13.2 versus 7.4 lb). Baseline weight and year-five weight in all race and gender groups were strongly associated, suggesting a high degree of tracking of adiposity during young adulthood. Greater weight gain was noted in participants reporting baseline education of high school or less versus college graduates in black women (14.4 versus 10.0 lb, p < 0.05), white women (10.2 versus 5.2 lb, p < 0.0001) and white men (10.2 versus 7.8 lb, p < 0.001). Significantly greater weight gain was observed in younger (18-24 years) versus older (25-30 years) men, but no age-related difference was seen in women. The racial differences in weight gain remained after adjustment for age and level of education. The above trends were confirmed for other measures of body size, i.e. body mass index and skinfold thickness. CONCLUSION: These data indicate that young adults are at high risk of weight gain, and that weight gain was greatest among African Americans and among less educated participants. These high-risk groups can be identified and targeted for primary prevention of adult obesity in addition to population wide efforts that will be required to counteract the secular trend of increased obesity observed in US adults. PMID- 9395578 TI - Ethnic differences in body composition and their relation to health and disease in women. AB - Differences in body composition between black and white women have been well established. Black women have more bone and muscle mass, but less fat, as a percentage of body weight, than white women, after controlling for ethnic differences in age, body weight, and height. In addition, black women have more upper-body fat than white women. These ethnic differences in body composition appear to be associated with disease risk in women. The greater skeletal and muscle mass in black compared to white women appears to protect them from osteoporosis. The relationship between fat distribution and cardiovascular disease also appears to be influenced by ethnicity. This review has two purposes: (1) To examine previous research investigating ethnic differences in body composition between black and white women; and (2) To demonstrate the relationship between body composition and disease in women as a function of ethnicity. PMID- 9395579 TI - Ethnicity, hypertension, coronary heart disease and renal diseases in South Africa. AB - This paper reviews the impact of race and environment upon hypertension, coronary heart disease and renal diseases in South Africa. Inequalities of socioeconomic status, lifestyle, and access to South African health care have produced striking differences in the prevalence and complications of hypertension. Coronary heart disease is 'epidemic' in the white and Indian population and is still relatively uncommon in blacks. There are different histological patterns of glomerulonephritis among the racial groups, which may lead to end-stage renal disease. Hypertension is an important cause of end-stage renal disease in the black population whilst analgesic nephropathy is important in the white population. Efforts are now being made to comprehend these daunting realities and to minimize the inequalities. PMID- 9395580 TI - Stigmatization, discrimination and fear of AIDS in Greece: implications for health policy. AB - A prospective health-education research project about AIDS knowledge and attitudes towards AIDS was conducted in Athens and nine adjacent municipalities in west Attica, Greece. Socioeconomic and demographic data, AIDS knowledge, and attitudinal information were collected from 1552 respondents and analysed treating the attitudes of stigmatization, discrimination and fear towards AIDS as the dependent variable. Statistically significant correlations were found between each of the three attitudinal variables and the independent ones; specifically, age, place of residence, marital status and level of AIDS knowledge. Our working hypothesis--that the higher the level of AIDS knowledge, the lower the level of discrimination and stigmatization--was supported by our data. The relationship between AIDS knowledge and fear was less clear. Fear probably inhibits a rational approach to screening for HIV, and more empirical research is needed about fear and its interaction with stigmatizing and discriminatory attitudes and behaviours. Such research should be aimed at identifying population groups 'at risk' of expressing high levels of negative social attitudes about AIDS so that educational programmes can be appropriately designed. PMID- 9395581 TI - Differential survival in blacks and Hispanics with AIDS. AB - OBJECTIVE: The object of this study was to investigate whether there are differences in survival by ethnicity in people with AIDS. DESIGN: The CDC Public Access Dataset was analysed. To estimate survival more accurately, a cohort of individuals diagnosed in 1987 was chosen from the dataset. Using this analysis, probabilities of survival were estimated. RESULTS: There were significant differences in survival in blacks and Hispanics as compared to whites diagnosed in 1987. Although there are differences in survival by transmission category, survival differences by ethnicity persisted when analysed within specific transmission categories. A model where the frequency distributions of survival were log-transformed suggests that disease progression per se may not be the most important factor, but time of diagnosis may be. In addition, in looking at median survival by year of diagnosis, it is clear that blacks and Hispanics have not shown the same magnitude of improvement in survival time, and lag behind whites. CONCLUSIONS: This study clearly shows differences in survival with AIDS by ethnicity. Differential access to health care may underlie such ethnic differences in survival. PMID- 9395582 TI - Language background and communication skills of medical students. AB - OBJECTIVE: The increasing proportion of medical students whose primary language is other than English and recent reports of poor communication skills of medical graduates has generated community concern about the methods of selection of students and their communication skills training. This paper examines the relationship between language background and examination performance in oral communication skills in Year 5 medical students. METHOD: Questionnaire data from all Year 5 students in the 1992 general practice terms were matched to examination results. RESULTS: Seventy percent of students responded. Most students whose primary language was not English passed, although some required remedial communication skills tuition. The most powerful predictors of poor performance were recent arrival in Australia and living in an environment where English was not spoken at home. CONCLUSION: Students with poor English oral communication skills should be encouraged to speak English away from the medical school and should be offered additional tuition so that their skills in other languages are not lost to the health-care system. PMID- 9395584 TI - Patterns of mortality among Bangladeshis in England and Wales. AB - OBJECTIVES: To investigate the patterns of mortality among Bangladeshis living in England and Wales. METHODS: An analysis of national mortality data, classified by country of birth, for the latest period (1988-1992), using the method of indirect standardization for deriving standardized mortality ratios (SMRs) with the age- and sex-specific rates for England and Wales as the standard (= 100). The SMRs were derived for Bangladeshi-born men and women aged 20-69 years for major disease entities. RESULTS: The mortality among Bangladeshi men was significantly higher (SMR 118 and 95% CI 111-126) than the levels prevalent in England and Wales. In contrast, the mortality among Bangladeshi women was significantly lower (SMR 71 and 95% CI 61-82). The cancer mortality overall was lower than expected in both sexes, with the exception of cancer of the liver and gall bladder. The mortality from breast cancer (SMR 16 and 95% CI 6-34) and cervical cancer (SMR 51 and 95% CI 14-131) was lower than expected. Bangladeshi men experienced high mortality from diabetes (SMR 685 and 95% CI 529-874), coronary heart disease (SMR 148 and 95% CI 134-163) and cerebrovascular disease (SMR 267 and 95% CI 222-319); they also experienced excess deaths from cirrhosis of the liver (SMR 254 and 95% CI 175-357). CONCLUSIONS: The findings establish significant variations in the recent health experiences of Bangladeshi men living in England and Wales, posing a major challenge for purchasers of care. If the Health of the Nation strategy is to ensure that equity in health and health care is to apply to all those living in this country, the Bangladeshi population needs special targeting. PMID- 9395585 TI - Cultural variation in health locus of control. AB - OBJECTIVE: To compare health locus of control scores in women from different ethnic backgrounds. METHOD: One-hundred and twenty-eight caucasian, South Asian and Afro-Caribbean women completed written or orally presented versions of the multidimensional health locus of control scale, as well as ratings of religiousness, health status and occupational status. RESULTS: South Asian women scored higher on 'chance' and 'powerful others' locus of control as predicted. They also had higher scores on internality. The ethnic differences persisted after controlling for occupation and health status. High religiousness among the South Asians appeared to explain some, but not all, of their higher scores. CONCLUSION: South Asians may differ from British caucasians in relation to their beliefs about internal and external influences on health. PMID- 9395586 TI - Has psychological distress among UK South Asians been under-estimated? A comparison of three measures in the west of Scotland population. AB - OBJECTIVES: Previous work has shown low levels of psychological distress among UK South Asians, but some argue that the distress is under-reported. The present paper assesses distress on one clinically validated measure (the 12-item General Health Questionnaire), a psychosomatic measure and a self-report measure. METHODS: Interviews of 159 South Asians in Glasgow aged 30-40 years, mean age 35 years and 319 from the general population, all aged 35 years. RESULTS: The three distress measures were moderately correlated and at the thresholds chosen there was no hierarchy of severity between them. Distress on the GHQ12 was at similar levels for all the social groups assessed, but distress on the psychosomatic measure and self-assessment was higher for women, Muslims and limited English speakers. CONCLUSIONS: Clinical measures may have under-estimated distress in several South Asian groups. The results may be due to a preference for a particular language of emotion in the affected groups or to a higher frequency of stressful situations which provoke distinctive reactions. PMID- 9395587 TI - Why are African Americans under-represented in medical research studies? Impediments to participation. AB - OBJECTIVES: In accordance with the NIH Revitalization Act of 1993, the National Institutes of Health and the Alcohol, Drug and Mental Health Administration require grant applicants and cooperative agreement participants to include minorities in human subject research. In an environment characterized by diminishing research dollars, this mandate has increased the pressure on investigators to determine factors that impede minority participation and to develop strategies to overcome these impediments. METHODS: An extensive review of the literature was conducted to identify the factors possibly responsible for the low participation levels of African Americans in medical research studies and to highlight areas for further research. The items examined included the historical relationship between African Americans and medical researchers and the attitudes, perceptions and beliefs of potential participants and researchers as they relate to the low representation of African Americans in medical research. RESULTS: The factors identified as possible impediments to African American participation included distrust of the medical/scientific community, poor access to primary medical care, the failure of researchers to recruit African Americans actively, the alienation of minority health professionals, lack of knowledge about clinical trials, language and cultural barriers. CONCLUSIONS: Well-designed, relevant, ethical research in conjunction with an appreciation of the many barriers to participation are paramount to increasing African American presence in clinical research. PMID- 9395588 TI - Lipoprotein (a) distribution in a Nigerian population. AB - OBJECTIVES: To determine the distribution and determinants of lipoprotein (a) (Lp(a)) concentration among Nigerians. METHODS: Subjects were recruited from civil servants living in Benin City, Nigeria. The height and weight of the individuals were measured and their use of alcohol and tobacco estimated by questionnaire. Laboratory analyses of blood samples involved Lp(a), total cholesterol (TC), high-density lipoprotein (HDLc), HDL2c, HDL3c, triglyceride (TG) and insulin. RESULTS: The analyses indicate that the Lp(a) concentrations are elevated among Nigerian populations and more skewed towards high levels than is observed for caucasian and oriental groups. The median levels for Lp(a) were 24.0 mg dl-1 and 19.0 mg dl-1 for women and men, respectively. This difference was significant (P < 0.05) but after stratifying by age, only the 45-54 year-old group of women (30.1 mg dl-1) had significantly higher (p < 0.001) median concentrations of Lp(a) than men (18.4 mg dl-1). Age, 20-64, had no influence on Lp(a) levels in men but in women Lp(a) concentrations increased significantly with age (p < 0.05). Among males alcohol consumption, smoking and body mass index (BMI) were not related to Lp(a) concentrations but a significant effect (p < 0.05) was noted for waist-hip ratio (WHR). Among females no relationship was observed between Lp(a) levels and alcohol consumption, BMI and WHR. All serum lipids measured (TC, HDLc, HDL2c, HDL3c, low-density lipoprotein (LDLc), and TG) were correlated with Lp(a) concentrations among men. A significant association with TC and LDLc remained after correcting for Lp(a) cholesterol. Among women, the Lp(a) levels were associated with TC, HDLc, HDL3c, and LDLc but not with HDL2c, and TG. The correlations with TC and LDLc were not significant after correcting for Lp(a) cholesterol. Insulin did not correlate with Lp(a) levels in either men or women. CONCLUSIONS: Lp(a) concentrations are high in Nigerians, particularly among women, and the association between the Lp(a) concentrations and other lipoproteins is stronger than in white populations. PMID- 9395589 TI - Knowledge, uptake and availability of health and social services among Asian Gujarati and white elderly persons. AB - OBJECTIVES: To investigate factors affecting the uptake of health and social services by elderly Asian Gujarati. METHODS: Four hundred and five Hindu Gujaratis and 381 whites aged 65 years and over residing in Leicester were randomly sampled from the Leicestershire District FHSA list by a computerized method based on linguistic analysis of the patient's name. One hundred and fifty Hindu Gujaratis and 152 whites were interviewed with response rates of 72% for the Asian Gujaratis and 80% for the white groups. The outcome measures were the activities of daily living (ADLs), incontinence, auditory/ visual deficits, cardiovascular disease, cognitive impairment (measured by the Mini-mental State Examination), depression, use of GP and hospital services, knowledge of community health and social services, willingness to use, suitability and cultural accessibility. RESULTS: The poorer uptake of services by elderly Asian Gujarati could not be explained by better health. They were significantly more likely to be dependent in six of the 14 ADLs and had higher rates of diabetes and impaired vision. Significantly more Asian Gujaratis than whites lived with others (84 versus 52%, p < 0.0001) with a greater availability of alternative sources of help and support. The knowledge and understanding of services were significantly poorer in the Gujarati group; fewer Asian Gujaratis knew how to apply for services and of those applying, fewer had been successful. Where services had been obtained, the levels of dissatisfaction were higher in the Gujarati group. The literacy rates were low in the Gujarati sample with 79% being unable to read or write in English and 27% unable to read or write in their mother tongue. CONCLUSIONS: The lower uptake of services by elderly Asian Gujarati is not the result of better health but may be explained by greater family support together with a lack of knowledge of and dissatisfaction with what is available. Health services will need to reappraise and revise some of their practices if they are to cater adequately for this growing population with many needs as yet unmet. PMID- 9395590 TI - Use of selected high-fat foods by Hispanic adults in the northeastern US. AB - OBJECTIVE: To add to the limited information of dietary fat intake of US Hispanic adults, in particular for subgroups other than Mexican Americans. METHODS: The frequency of eating 13 high-fat food items commonly consumed in the US was examined in 665 Hispanic adults 20-74 years old in Connecticut and Long Island, New York, sampled from Spanish-surname telephone listings and surveyed by telephone in 1992. RESULTS: Mean estimated fat intake from the 13 items was significantly greater for the 357 men than the 308 women; the largest gender differences were for hamburgers/cheeseburgers and french fries. Whole milk was an important contributor to the fat intake of persons with the highest fat intakes. In multiple linear regression analyses, age (negative association) and gender, but not education and acculturation (based on language spoken, read and written), were statistically significant predictors of fat intake from the 13 items. CONCLUSIONS: Longitudinal studies using diet diaries are needed in these Hispanic populations. PMID- 9395591 TI - Epidemiology of self-reported past heavy drinking in Hispanic adults. AB - OBJECTIVES: Self-reports of past heavy drinking correlate with the current drinking practices and with risk of mortality in non-Hispanic males. The prevalence of past heavy drinking has not been reported in Hispanic populations. METHODS: Using data from the Hispanic Health and Nutrition Examination Survey (HHANES) we (1) report on the prevalence, duration and severity of past heavy drinking in three Hispanic groups, (2) compare the current alcohol consumption patterns among past heavy drinkers and those who do not report a history of past heavy drinking and (3) compare the risk factor profiles and health indicators in these two groups. RESULTS: The prevalence of past heavy drinking among Mexican American and Puerto Rican males ranged from 28-35% while the rates for Cuban American males ranged from 7-16%. The rates for Hispanic women were much lower (1 8%). The average years of past heavy drinking ranged from 2.3-14.9 years, while the alcohol consumption during the past heavy drinking period ranged from 24.4 44.0 drinks per week. Past heavy drinkers tended to consume more alcohol at present than did never heavy drinkers with the greatest differences found for Mexican American females. Comparisons of the risk factors and health indicators by drinking status revealed a higher prevalence of smoking among past heavy drinkers (50-60%) versus never heavy drinkers (34-43%). Past heavy drinking Mexican American females also reported significantly more chronic conditions and depressive symptoms than did never heavy drinkers. CONCLUSIONS: Prevalence rates of past heavy drinking among Mexican American and Puerto Rican males are approximately three times higher than rates reported for non-Hispanic male populations. PMID- 9395592 TI - Longitudinal effects of an HIV testing and counseling programme for low-income Latina women. AB - OBJECTIVES: The purpose of this study was to assess the effects of an HIV antibody testing, counseling and education programme on the knowledge and practices of low-income Los Angeles Latina women. METHODS: The study design was prospective and longitudinal involving pre-test, post-test and retest measures over a 2-year period. The study employed an experimental group and a comparison group which did not receive the intervention. The study group was comprised of a convenience sample of 508 low-income Latina women who were recruited from the Public Health Service nutrition programme for women, infants and children (WIC). The comparison group (n = 51) was recruited from the same setting. A battery of instruments was selected to measure HIV knowledge and practices, the social support received, self-esteem, the level of acculturation and sociodemographic characteristics. The instruments were administered at pre-test, 2 weeks post-test and 1 year retest. The HIV antibody serostatus was assessed at pre-test and retest. An intervention protocol based on cultural competence, women as traditional health care givers and the major transmission categories was provided after the pre-test and was reinforced post-test. Finally, qualitative data were collected from the focus group participants (n = 55) to evaluate the intervention protocol. RESULTS: The participants in the study made significant improvements in HIV knowledge and reported condom use practices from pre-test to post-test that were retained on retest. The comparison group subjects did not make significant pre-test-post-test improvements on these measures. CONCLUSIONS: It should be noted that the changes in practices made by the study group did not necessarily reduce their risk of HIV infection or transmission and were not related to the demonstrated knowledge and skills improvement. Of special significance to programme planners, educators and researchers, both the quantitative and qualitative data revealed problem areas with the intervention protocol related to cultural norms and the possible fragmentation of information based on the behavioral transmission categories. PMID- 9395593 TI - Public policy analysis of Indiana's minority health initiatives. AB - Similar to national health trends in the US, racial/ethnic minorities in the state of Indiana continue to experience disparities in poor health status from preventable health conditions. To address this problem, people from minority communities across the state mobilized a broad base of individuals and organizations to facilitate the successful legislative enactment of a statewide minority health initiative. A case study of the initiative is presented for public policy analysis. The theoretical framework for the study is Etzioni's Societal Guidance Theory. The findings show that minority health advocates were able to impact favorably on public policy formulation and funding of the initiative by increasing knowledge about minority health status among grass-roots people, generating public consensus for public policy intervention, setting mutual goals via a 5-year strategic minority health plan, creating organizational structures to implement the plan and utilizing power to push the initiative through the legislative process. The weaknesses of the initiative efforts include a limited infrastructure development of minority health coalitions, restricted effective use of the legislative process and varying degrees of linkages among other advocacy groups. Improvements in these areas are discussed and recommendations are made for the implementation phase of the initiative. PMID- 9395594 TI - Racial differences in post-neonatal mortality in Chicago: what risk factors explain the black infant's disadvantage? AB - OBJECTIVES: To investigate the extent to which the place of residence affects the black to white differential in post-neonatal (28-365 days) mortality, we performed a univariate analysis and multivariate logistic regression of the 1982 1983 Illinois vital records. Chicago Police violent crime information and 1980 US Census income data. METHODS: Four environmental predictors of post-neonatal death were examined: a median family income of < $10,000 per year, a poverty prevalence of > 50%, violent crime rates of > 11/1000 and limited community access to primary medical care based on physician supply ratios. RESULTS: The post-neonatal mortality rate of black (n = 50,765) infants was three times that of white (n = 50,690) infants: 10/1000 versus 3/1000, respectively. Thirty-six percent of the white infants had none of the environmental risk factors, whereas only 13% of the black infants had none of the risk factors. For black infants, the presence of any one factor was associated with a slightly increased risk of post-neonatal mortality (9/1000 as compared to 7/1000 with no risk factors), whereas the presence of two or more risk factors was associated with a higher risk (11/1000). When the number of these environmental risk factors were taken into account, the OR for black infants declined from 3.0 (95% CI 2.5-3.6) to 1.7 (95% CI 1.5-1.9). When the differences in maternal age, education, marital status and infant birth weight were also taken into account the odds ratio of post-neonatal death for blacks was 1.5 (95% CI 1.3-1.7). CONCLUSIONS: We conclude that a substantial proportion of the black to white difference in post-neonatal mortality is associated with specific environmental conditions. PMID- 9395596 TI - Alternative and complementary medicine: part of human heritage. PMID- 9395597 TI - The importance of alternatives to health care reform. PMID- 9395595 TI - Cardiovascular and plasma catecholamine response to static exercise in normotensive blacks and whites. AB - OBJECTIVES: The objectives of the present study were (1) to evaluate the pressor response to an isometric handgrip exercise in normotensive black and white males; (2) to measure plasma catecholamine levels pre- and post-exercise, as an index of sympathetic nervous system activity; and (3) to quantify the pressor response to bolus intravenous injections of phenylephrine (an alpha-specific agonist). METHODS: Cardiovascular and catecholamine responses to an isometric handgrip exercise (3 minutes at 30% MVC) were measured in 15 normotensive blacks and whites. In another phase of the study, pressor responses to bolus injections of phenylephrine were assessed to evaluate alpha-adrenergic sensitivity. RESULTS: The blood pressure in the blacks increased from 119/69 to 160/120 mm HG during isometric exercise, while in the whites it increased from 118/67 to 153/110 mm HG. The blacks exhibited a greater diastolic blood pressure reactivity, as evidenced by a significant race x time interaction (p < 0.05). The heart rate responses were not significantly different between the two groups. The plasma levels of norepinephrine were similar at rest, but were 25% lower in the blacks than in the whites following isometric exercise (p < 0.01). Black subjects also demonstrated an increased pressor response to intravenous injections of phenylephrine at rest (p < 0.05). CONCLUSIONS: The enhanced vascular sensitivity to norepinephrine may have contributed to the greater exercise pressor response in the blacks. PMID- 9395598 TI - Assessment of plants as medicines: a tale of two tales. PMID- 9395599 TI - The regulation of acupuncture needles by the United States Food and Drug Administration. PMID- 9395600 TI - An image of distant contact: a blind Japanese massage practitioner. PMID- 9395601 TI - Social science theory and methods in the study of alternative and complementary medicine. AB - Anthropology is a holistic science with theory, data, and methods that can be of great service to researchers on alternative medicine. In this paper useful models and methodologic stances were identified that can help researchers to deal creatively with the stresses imposed on science by worldview preferences that differ among both scientists and healthcare systems. I have argued that rather than prefer one paradigm over another, researchers should select techniques based on a rationale featuring deep knowledge of the context of the healthcare issue they want to study. This will not only produce the most accurate and useful data, but should also help free science of its current strictures and allow expansion into a wider conversation about human and medical realities. PMID- 9395602 TI - Social context of complementary medicine in Western society, Part I. PMID- 9395603 TI - A critical analysis of acupuncture in pulmonary disease: efficacy and safety of the acupuncture needle. AB - Criteria for therapeutic efficacy and safety include significant amelioration of symptoms and, ideally, cure (i.e., patients' belief in effective improvement of symptoms and quality of life, durable impact on symptoms, verifiable subjective and objective changes); improved patient management (e.g., diminishing, or ceasing medication, physiotherapy, and other interventions); safety for patient and practitioner and an acceptable side effect profile; cost-effectiveness of the therapy in practice and to teach to others. There is evidence that in bronchial asthma, chronic bronchitis, and chronic disabling breathlessness the use of acupuncture fulfills these to varying degrees. It can facilitate reducing pharmacologic medication and is safe, suggesting that acupuncture as an adjuvant in the treatment of respiratory disease might be safer than prolonged pharmaceutical maintenance therapy alone. Its cost-effectiveness has yet to be adequately researched. Twenty-one papers in English were obtained and 16 were further evaluated; eight were double-blind, five single-blind, and three unblinded. The remaining five, and most of the Chinese literature, were excluded on account of their poor quality. Acupuncture was effective in four of eight of the double-blind, three of five single-blind, and three of three unblinded studies (i.e., 10 of 16 [62.5%] overall). A previously unreported confounding variable was identified and concerned the designation of sham points. Most sham points were believed to be inactive but, according to traditional Chinese principles, many are active in pulmonary disease. Reappraised accordingly, the unequivocally positive studies were summed with those in which "real" and "sham" acupuncture were not significantly different but in which the combined effect of all acupuncture (i.e., real + sham) on breathlessness was significantly different from baseline. This yielded 13 of 16 (81%) [corrected] studies in which acupuncture led to significant improvement. In most studies, current pharmacologic treatment had a greater effect than acupuncture alone. However, in the 11 studies in which it was evaluated, medication could be significantly reduced by acupuncture in 10 (91%). Twenty-three of the 320 patients in the 16 studies (7%) reported minimal side effects, none requiring intervention. Current published evidence reveals no reason to withhold acupuncture as a safe and potentially effective treatment in patients with bronchial asthma and chronic obstructive lung disease. Further, more appropriately designed studies are urgently required. This would be facilitated in the United States by licensing the acupuncture needle as a therapeutic agent and might lead to important new insights and therapeutic opportunities. PMID- 9395604 TI - Treatment of seasonal affective disorder with a high-output negative ionizer. AB - This study was designed to evaluate the antidepressant effect of negative ions in the ambient air as a potential treatment modality for seasonal affective disorder. Twenty-five subjects with winter depression underwent a double-blind controlled trial of negative ions at two exposure densities, 1 x 10(4) ions/cm3 or 2.7 x 10(6) ions/cm3, using an electronic negative ion generator with wire corona emitters. Home treatments were taken in the early morning for 30 min over 20 days, followed by withdrawals. The severity of depressive symptoms (prominently including the reverse neurovegetative symptoms of hypersomnia, hyperphagia, and fatigability) decreased selectively for the group receiving high density treatment. Standard depression rating scale assessments were corroborated by clinical impressions. When a remission criterion of 50% or greater reduction in symptom frequency/severity was used, 58% of subjects responded to high-density treatment while 15% responded to low-density treatment (chi 2 = 5.00, df = 1, p = 0.025). There were no side effects attributable to the treatment, and all subjects who responded showed subsequent relapse during withdrawal. Treatment with a high-density negative ionizer appears to act as a specific antidepressant for patients with seasonal affective disorder. The method may be useful as an alternative or supplement to light therapy and medications. PMID- 9395605 TI - Introducing alternative/complementary healing to allopathic medical students. AB - We have designed a senior elective, Introduction to Alternative Medicine, to prepare our students better to practice in multicultural environments, and to expand their views of health and healing. We combined didactic lecture, films, first-hand experience with some methods, and observation of alternative practitioners in their offices/clinics. Students explored hypnosis, chiropractic, therapeutic touch, meditation, biofeedback, acupuncture, homeopathy, naturopathy, and massage therapy. Discussions of scientific efficacy, legal and ethical considerations, and the role of spirituality in health and healing focused on limitations of science-based approaches and reasons why alternative/complementary methods are popular with patients and allopathic physicians. We conclude that allopathic medical schools have an important role in reducing the isolation of their students from alternative health beliefs, practices, and systems of care that are common in our communities. PMID- 9395606 TI - Characteristics of complementary medical systems: the ubiquitous use of plants. PMID- 9395607 TI - Alternative medicine. PMID- 9395608 TI - Incense over a medicine bundle: Hidatsa. PMID- 9395609 TI - Plants, food and civilization: the lessons of indigenous Americans. PMID- 9395610 TI - The role of plants in traditional medicine and current therapy. AB - Recent years have witnessed a renewed interest in plants as pharmaceuticals in the Western world. This interest is channeled into the discovery of new biologically-active molecules by the pharmaceutical industry and into the adoption of crude extracts of plants for self-medication by the general public. In both of these areas some attention is being paid to the investigation and use of ethnopharmacology, the traditional use of plants for medicinal purposes by particular cultural groups. Ethnopharmacologic leads have resulted in the introduction of new single molecule drugs but have a greater role to play if crude extracts are accepted for clinical use in the West. The problems confronting such usage are discussed. Considerable benefits for developing countries are possible when the local medicinal plants are subjected to scientific methods of validation of traditional use and quality control. This approach has met with success in some parts of the world but is not always appreciated by national governments and international agencies. Related areas of concern such as conservation of ecology and culture must be integrated with any such program. Plants used in traditional medicine therefore have an important role to play in the maintenance of health in all parts of the world and in the introduction of new treatments. PMID- 9395611 TI - Results of five randomized studies on the immunomodulatory activity of preparations of Echinacea. AB - This article describes and discusses five placebo-controlled randomized studies investigating the immunomodulatory activity of preparations containing extracts of Echinacea in healthy volunteers. A total of 134 (18 female and 116 male) healthy volunteers between 18 and 40 years of age were studied. Two studies tested intravenous homeopathic complex preparations containing Echinacea angustifolia D1 (study 1) and D4 (study 5). Two studies (2 and 3a) tested oral alcoholic extracts of roots of E. purpurea, one study an extract of E. pallida roots (study 3b), and one study an extract of E. purpurea herb (study 4). Test and placebo preparations were applied for four (study 5) or five (studies 1-4) consecutive days. The primary outcome measure for immunomodulatory activity was the relative phagocytic activity of polymorphonuclear neutrophil granulocytes (PNG), measured in studies 1 and 2 with a microscopic method and in studies 3, 4, and 5 with two different cytometric methods. The secondary outcome measure was the number of leukocytes in peripheral venous blood. Safety was assessed by a screening program of blood and other objective parameters as well as by documentation of all subjective side effects. In studies 1 and 2 the phagocytic activity of PNG was significantly enhanced compared with placebo [maximal stimulation 22.7% (95% confidence interval 17.5-27.9%) and 54.0% (8.4-99.6%), respectively], while in the other studies no significant effects were observed. Analysis of intragroup differences revealed significant changes in phagocytic activity during the observation periods in five test and three control groups. Leukocyte number was not influenced significantly in any study. Side effects due to the test preparations could not be detected. Our studies provide evidence for immunomodulatory activity of the homeopathic combination tested in study 1 and the E. purpureae radix extract tested in study 2. The negative results of the other three studies are difficult to interpret due to the different methods for measuring phagocytosis, the relevant changes in phagocytic activity within most placebo and treatment groups during the observation period, and the small sample sizes. Future studies should be performed on patients rather than healthy volunteers and use standardized or chemically defined monopreparations of Echinacea. PMID- 9395612 TI - Social context of complementary medicine in Western society, Part II: traditional Chinese medicine and HIV illness. PMID- 9395613 TI - A systems theory approach to an expanded medical model: a challenge for biomedicine. PMID- 9395614 TI - A model for a networked information resource on alternative medicine. PMID- 9395615 TI - The NIH Methodology Conference: the methodology debate in the United Kingdom during the past ten years. PMID- 9395616 TI - Acupuncture and stroke rehabilitation. PMID- 9395617 TI - Social science theory and methods in the study of alternative and complementary medicine. PMID- 9395618 TI - Women chiropractors, 1921. PMID- 9395619 TI - Traditional health systems: policy, biodiversity, and global interdependence. PMID- 9395620 TI - Phytomedicines: the greening of modern medicine. AB - Although available in the United States for little more than 10 years, phytomedicines represent an important class of natural therapeutics that possess significant scientific literature as well as broad use and acceptance worldwide. Until recently, such products were severely limited in the United States. Passage of the DSHEA has opened important new opportunities to communicate the use and benefits of phytomedicines as safe and useful natural health care products. Broader dissemination of the extant scientific literature on phytomedicines can and should have an important effect on the view of the conventional medical community that plant extracts specifically, and natural products generally, offer valuable and needed benefits to America's aging population, as well as to others. Although used since ancient times, plants are enjoying a renaissance of interest and use in the United States. As Pliny the Younger said, "Everything old is new again." PMID- 9395621 TI - An alternative medicine treatment for Parkinson's disease: results of a multicenter clinical trial. HP-200 in Parkinson's Disease Study Group. AB - The natural occurrence of antiparkinsonian drugs in plants--anticholinergics in Datura stramonium, levodopa in Mucuna pruriens and Vicia faba, dopamine agonist activity in Claviceps purpura, and MAO inhibitor activity in Banisteria caapi-are known. Our study examined the efficacy and tolerability of HP-200, derived from Mucuna prurient, in patients with Parkinson's disease. Sixty patients with Parkinson's disease (46 male and 14 female) with a mean (+/- SD) age of 59 +/- 9 years were treated in an open study for 12 weeks. Of these, 26 patients were taking synthetic levodopa/carbidopa formulations before treatment with HP-200, and the remaining 34 were levodopa naive. HP-200, a powder (supplied as a 7.5 g sachet), was mixed with water and given orally. The Unified Parkinson's Disease Rating Scale (UPDRS) was used at baseline and periodically during the 12-week evaluation. Statistically significant reductions in Hoehn and Yahr stage and UPDRS scores were seen from baseline to the end of the 12-week treatment (p < 0.0001, t-test). The group mean (+/- SD) dose for optimal control of symptoms was 6 +/- 3 sachets. Adverse effects were mild and were mainly gastrointestinal in nature. No adverse effects were seen in clinical laboratory reports. HP-200, developed from an alternative medicine source, Ayurveda, was found to be an effective treatment for patients with Parkinson's disease. PMID- 9395622 TI - Electroacupuncture reduces morphine-induced emesis in ferrets: a pilot study. AB - We treated morphine-induced emesis in ferrets with electroacupuncture (EA) for five minutes at the acupuncture point Neiguan (P6) using two different frequencies of stimulation: 1.0 Hz and 5.0 Hz (n = 5 ferrets per group). Both EA treatments reduced the number of emetic episodes (39% reduction of episodes, p < or = 0.05 at 1.0 Hz; 43% reduction of episodes, p < or = 0.05 at 5.0 Hz) and prolonged the onset time of emesis as compared with control receiving no EA. This model demonstrates that under these conditions EA is effective as an antiemetic against morphine in the ferret and may allow for further studies on the mechanisms of acupuncture and emesis. PMID- 9395623 TI - Stress reduction and preventing hypertension: preliminary support for a psychoneuroendocrine mechanism. AB - Our objective was to identify endocrine-related mechanisms capable of mediating preventive effects of stress reduction in hypertensive heart disease. Since beneficial effects of stress reduction accrue over time, this cross-sectional, descriptive study sought differences between healthy students not practicing a systematic technique for reducing stress (the average stress, or AS, group, n = 33) and a similar group who for 8.5 years had practiced the Transcendental Meditation (TM) technique, used widely to reduce stress (the low stress, or LS, group, n = 22). The two groups of students, matched for age and area of study, performed timed collections of urine that included (separately) the entire waking and sleeping portions of 1 day. They also completed the Profile of Mood States and the State-Trait Anxiety Inventory, self-report instruments sensitive to subjective level of stress. Urine samples were analyzed for adrenocortical steroids by radioimmunoassay, for Na+, K+, Mg2+, Ca2+, and Zn2+ by atomic absorption spectrometry, and for neurotransmitter metabolites by reverse-phase, high-performance, liquid chromatography, and spectrophotometry. The two groups differed significantly on most measures. Specifically, the LS group was lower in cortisol and aldosterone and higher in dehydroepiandrosterone sulfate (DS) and the serotonin metabolite, 5-hydroxyindoleacetic acid (5-HIAA). Excretion of sodium, calcium, zinc, and the norepinephrine metabolite, vanillylmandelic acid (VMA), was also lower in this group, as were Na+/K+ ratio, mood disturbance, and anxiety. In women practicing TM, cortisol correlated inversely and DS directly with number of months of TM practice. The results identify improvements in mood state, adrenocortical activity, and kidney function as probable factors in the preventive and treatment effects of stress reduction. Because suboptimal levels of these parameters result from chronic, subjective stress, the findings add mechanistic support to the contention that hypertensive heart disease is avoidable, even in modern industrialized societies. PMID- 9395624 TI - The effects of self-hypnosis on quality of life following coronary artery bypass surgery: preliminary results of a prospective, randomized trial. AB - The effects of complementary techniques and alternative medicine on allopathic therapies is generating much interest and research. To properly evaluate these techniques, well controlled studies are needed to corroborate the findings espoused by individuals practicing complementary medicine therapies. To this end, we evaluated the role of one of these therapies, self-hypnosis relaxation techniques, in a prospective, randomized trial to study its effects on quality of life after coronary artery bypass surgery. Subjects were randomized to a control group or a study group. Study group patients were taught self-hypnosis relaxation techniques the night prior to surgery. The control group received no such treatment. Patients then underwent routine cardiac management and care. The main endpoint of our study was quality of life, assessed by the Profile of Moods Scale. Results demonstrated that patients undergoing self-hypnosis the night prior to coronary artery bypass surgery were significantly more relaxed than the control group (p = 0.0317). Trends toward improvement were also noted in depression, anger, and fatigue. This study demonstrates the beneficial effects of self-hypnosis relaxation techniques on coronary surgery. This study also identifies endpoints and a study design that can be used to assess complementary medicine therapies. Results of this preliminary investigation are encouraging and demonstrate a need for further well-controlled studies. PMID- 9395625 TI - NIH Office of Alternative Medicine Conference: federal agencies explore the potential role of botanicals in U.S. health care. PMID- 9395626 TI - Homeopathic physician, 1855. PMID- 9395627 TI - The power of dance: health and healing. AB - Dance involves the culturally mediated body, emotion, and mind. So do illness and pain. Dance may promote wellness by strengthening the immune system through muscular action and physiological processes. Dance conditions an individual to moderate, eliminate, or avoid tension, chronic fatigue, and other disabling conditions that result from the effects of stress. Dance may help the healing process as a person gains a sense of control through (1) possession by the spiritual in dance, (2) mastery of movement, (3) escape or diversion from stress and pain through a change in emotion, states of consciousness, and/or physical capability, and (4) confronting stressors to work through ways of handling their effects. PMID- 9395628 TI - Hospital 2000: the interface of radiology within a combined "complementary allopathic" medicine framework. AB - The field of radiology can play a crucial role in the movement beyond allopathic and complementary medicine to a true combination approach. Radiologists are proponents of minimal invasiveness and are familiar with the overreliance on diagnostic imaging tests. Their experience can be used to design appropriate applications for technology and to plan research methods to explore the questions raised by a combination medicine paradigm. PMID- 9395629 TI - "Self" and addiction: the role of imagery in self-regulation. AB - In this paper we discuss the role of self-regulation and self-perception in sustaining addictive behavior. We suggest that self-regulation techniques are unlikely to have a sustained positive effect on changing addictive behavior in the absence of a change in "addict" self-perception. We describe treatment approaches that emphasize the use of guided imagery for facilitating a shift in self-perception that may be more compatible with self-regulation. PMID- 9395630 TI - The health beliefs and behaviors of three groups of complementary medicine and a general practice group of patients. AB - Patients (n = 256), consulting either a general practitioner (GP) or one of three complementary practitioners (osteopath, homeopath, or acupuncturist), completed a seven-part questionnaire that looked at demographic data, medical history, familiarization with complementary therapies, health beliefs and life-style, health locus of control, scientific health beliefs, and their perceptions of the consultation style of general and complementary practitioners. The four subject groups did not differ significantly on the demographic variables of sex, years of schooling, whether or not they had a degree, marital status, or income, but did differ on age and number of children. The effects of both the significant demographic variables and some aspects of patients medical history were controlled for in subsequent analyses. Acupuncture patients stood out as having the most different chronic medical history. They were also least satisfied with their GP, had least confidence in prescribed drugs, and were most concerned with leading a healthy life-style. The acupuncture patients were most skeptical about orthodox medicine. The main finding was that patients of complementary practitioners are not a homogeneous group, but do differ in their views on satisfaction with GPs, healthy life-style, global environmental issues, confidence in prescribed drugs, faith in medical science, importance of a "healthy mind," harmful effects of medical science, and scientific methodology. The results imply that patients consult different practitioners, general or alternative, on the basis of a combination of their level of skepticism about orthodox medicine, their life-style, and other health beliefs. To talk of patients of complementary practitioners as a homogeneous group is fundamentally wrong. PMID- 9395632 TI - A patient-centered paradigm: a model for chiropractic education and research. AB - Within the chiropractic profession there is concern over both the appropriate paradigm for educating the student and the appropriate research model for investigating health care and promotion. The purpose of this study was to identify, interpret, and describe a paradigm for chiropractic education and research. Chiropractic first principles and existing health care paradigms were identified and integrated with the characteristics of chiropractic practice described in sociological studies of the chiropractic profession. A patient centered paradigm emerged, incorporating the principles of vitalism, holism, humanism, conservatism, naturalism, and rationalism. Characteristics of a patient centered paradigm that were identified were then subjected to an eight-member consensus panel with representatives drawn from chiropractic education, research, and sociology. The characteristics of a patient-centered paradigm agreed upon include self-healing, recognition of the patient as a unified whole, respect for the patient's values, beliefs, and dignity, involvement of the patient as a partner in health promotion, and a natural and conservative approach to evidence based care. Patient-centered research must reach beyond the randomized controlled trial, involving designs where clinicians apply their own patient-centered therapy in a "real world" assessment. A pluralism of methods, including both qualitative and quantitative studies, needs to be designed and implemented. Patient-centered research is a process that is pragmatic, realistic, and grounded in the day-to-day experiences of both patients and chiropractors. A patient centered paradigm offers a useful model to critically study what benefits patients and to prepare chiropractic students to practice in the patient's interest. PMID- 9395631 TI - Inhibition of several strains of influenza virus in vitro and reduction of symptoms by an elderberry extract (Sambucus nigra L.) during an outbreak of influenza B Panama. AB - A standardized elderberry extract, Sambucol (SAM), reduced hemagglutination and inhibited replication of human influenza viruses type A/Shangdong 9/93 (H3N2), A/Beijing 32/92 (H3N2), A/Texas 36/91 (H1N1), A/Singapore 6/86 (H1N1), type B/Panama 45/90, B/Yamagata 16/88, B/Ann Arbor 1/86, and of animal strains from Northern European swine and turkeys, A/Sw/Ger 2/81, A/Tur/Ger 3/91, and A/Sw/Ger 8533/91 in Madin-Darby canine kidney cells. A placebo-controlled, double blind study was carried out on a group of individuals living in an agricultural community (kibbutz) during an outbreak of influenza B/Panama in 1993. Fever, feeling of improvement, and complete cure were recorded during 6 days. Sera obtained in the acute and convalescent phases were tested for the presence of antibodies to influenza A, B, respiratory syncytial, and adenoviruses. Convalescent phase serologies showed higher mean and mean geometric hemagglutination inhibition (HI) titers to influenza B in the group treated with SAM than in the control group. A significant improvement of the symptoms, including fever, was seen in 93.3% of the cases in the SAM-treated group within 2 days, whereas in the control group 91.7% of the patients showed an improvement within 6 days (p < 0.001). A complete cure was achieved within 2 to 3 days in nearly 90% of the SAM-treated group and within at least 6 days in the placebo group (p < 0.001). No satisfactory medication to cure influenza type A and B is available. Considering the efficacy of the extract in vitro on all strains of influenza virus tested, the clinical results, its low cost, and absence of side effects, this preparation could offer a possibility for safe treatment for influenza A and B. PMID- 9395633 TI - Exploring homeopathic resources on the Internet: HOMEOWEB. AB - Electronic mail and other forms of communication on the Internet are especially important for complementary medicine disciplines, like homeopathy, which lack an efficient academic network for research purposes. The main advantages of using the Internet for homeopathy are outlined in this article. The first step to developing an electronic communications network was to create a Homeopathic Internet Resources List, which is available on-line now, and is being updated constantly. This references the great homeopathic libraries and reference databases. Most of these have been made accessible electronically through the Internet or by e-mail, although many homeopathic physicians are still unaware of the power of Internet communication. Since the summer of 1995 the HOMEOWEB webserver has tried to bring together scientific information on all aspects of homeopathic therapy on the Internet. This service will be enlarged in the near future, and the aim is to promote the Internet as the primary means of worldwide communication among the homeopathic community, ensuring that homeopathy survives into the next century. PMID- 9395634 TI - Legal ramifications of homeopathy. AB - The article addresses four regulatory challenges faced by practitioners of homeopathy: (1) medical practice acts, which prohibit the unlicensed practice of "medicine," (2) scope of practice limitations, which restrict nonmedical providers' ability to diagnose and treat disease; (3) prohibitions against "unprofessional conduct;" and (4) malpractice rules. The article concludes with suggestions for regulatory reform. PMID- 9395635 TI - National Institutes of Health Office of Alternative Medicine-Food and Drug Administration Workshop on Acupuncture. PMID- 9395636 TI - A short history of acupuncture. PMID- 9395637 TI - Acupuncture techniques and devices. PMID- 9395638 TI - Safety issues in acupuncture. PMID- 9395639 TI - Educational and licensing requirements for acupuncturists. PMID- 9395640 TI - Testimony to the Office of Alternative Medicine workshop on acupuncture. PMID- 9395641 TI - Educational and licensing requirements for medical practitioners. PMID- 9395642 TI - Clinical trials in support of medical devices. AB - 1. Clearly specify the hypothesis to be investigated using clinically meaningful objective endpoints. 2. Collect all patients' data in a reliable and consistent manner. 3. Use statistically appropriate procedures to analyze the data. 4. Conclusions should be able to support all claims of safety and effectiveness that are to be made. PMID- 9395643 TI - Scientific research into acupuncture for the relief of pain. PMID- 9395644 TI - Auricular acupuncture in animals: effects of opiate withdrawal and involvement of endorphins. PMID- 9395646 TI - Review of medical and clinical literature. PMID- 9395645 TI - Veterinary clinical applications of acupuncture. PMID- 9395647 TI - On the evaluation of the clinical effects of acupuncture: a problem reassessed and a framework for future research. PMID- 9395648 TI - Acupuncture in the treatment of pain. PMID- 9395649 TI - Overview of acupuncture in chronic pain clinical research. PMID- 9395650 TI - Review of acute and chronic pain published studies. PMID- 9395652 TI - Acupuncture as an antiemetic treatment. PMID- 9395651 TI - Overview of substance abuse acupuncture treatment research. AB - The research on the efficacy of acupuncture substance abuse treatment is generally still in an early stage. The methodological weaknesses found in the acupuncture research can be found in most substance abuse research. Sufficient early trial, empirical findings suggest that there are positive treatment effects. Certainly, use of the treatment is popular and widespread. Overall, the research has progressed beyond early clinical trials, and the method has been documented to be safe and potentially useful. PMID- 9395653 TI - Acupuncture in asthma and pulmonary disease: an analysis of efficacy and safety. PMID- 9395654 TI - Acupuncture in the treatment of paralysis due to central nervous system damage. PMID- 9395656 TI - Adverse effects of acupuncture. PMID- 9395655 TI - History of the Food and Drug Administration's regulation of acupuncture devices. PMID- 9395657 TI - South Vietnamese healing ceremony. PMID- 9395658 TI - U.S. patent documents on the Internet: an information resource for research in alternative and complementary medicine. AB - The US patent literature is a valuable source of information and research data relevant to studies in alternative and complementary medicine. Many patent publications contain detailed historical reviews, case reports, clinical trials, pilot studies, and basic biological research, any of which may give scientific insights, notwithstanding the controversies surrounding the issues of some patents, such as those of products of traditional medicinal plants. Much of the scientific, efficacy, and safety data necessary to obtain a patent is unpublished elsewhere. This paper gives an account of the recent experiment to make the full text of U.S. patents documents openly and freely available on the Internet, gives an example of browsing such a document from the AIDS Patents database, and provides examples of searching on-line on the Patents Abstracts database together with a miscellany of recent abstracts relevant to alternative and complementary medicine. PMID- 9395659 TI - Beneficial effect of Aloe on wound healing in an excisional wound model. AB - Recent evidence from in vitro and in vivo experiments suggests that topical antimicrobials may be toxic to fibroblasts and keratinocytes and retard wound healing. The purpose of this study was to determine the effects of Aloe, a potential wound-healing agent, on wound contraction in excisional wounds treated with topical antimicrobials. Sprague-Dawley rats were prepared with four 1.5 cm2 dorsal defects through the skin and panniculus. The animals were divided into five groups (n = 10 per group): (1) Aloe, (2) NaOCl solution (0.025%), (3) mafenide acetate, (4) mafenide acetate + Aloe, and (5) control. Wounds were treated topically for 14 days 3 times a day. Serial standard photographs and serial wound planimetry were performed weekly. Following healing, the breaking strength of each resultant scar was determined using an Instron tensiometer. Kruskal-Wallis, ANOVA, and multiple comparison methods were used for data analysis. Aloe and NaOCl solution significantly accelerated wound contraction (p < 0.05). In the mafenide acetate + Aloe group, contraction was similar to the control, whereas the mafenide acetate alone retarded wound healing. The addition of Aloe in combination and alone in wounds increased the breaking energy when compared to controls (p < 0.05). Aloe appears to expedite wound contraction and neutralize the wound retardant effect seen with the topical mafenide acetate alone. This effect appears to be due to an increased collagen activity, which is enhanced by a lectin, consequently improving the collagen matrix and enhancing the breaking strength. PMID- 9395661 TI - Adverse effects of acupuncture: a study of the literature for the years 1981 1994. AB - This study presents the adverse effects of acupuncture as recorded in the Medline database for the years 1981-1994. A total of 125 papers were localized by the keywords acupuncture adverse effects. Articles without case reports were excluded, and 78 reports forms the basis for the present paper. A total of 193 patients were reported with adverse effects of acupuncture in 14 years. Pneumothorax is the most common mechanical organ injury, while hepatitis dominates among infections. Acupuncture treatment is claimed to be responsible in the death of three patients. One patient died from bilateral pneumothorax, another got endocarditis, and died of complications. The third patient died of severe asthma while under acupuncture treatment. Most adverse effects of acupuncture seem to rely on insufficient basic medical knowledge, low hygienic standard, and inadequate acupuncture education. The study confirms the adverse effects of acupuncture under certain circumstances. Serious adverse effects, however, are few, and acupuncture can generally be considered as a safe treatment. PMID- 9395660 TI - Characterization of differing effects caused by homeopathically prepared and conventional dilutions using cytochrome P450 2E1 and other enzymes as detection systems. AB - Determination of cytochrome P450 2E1 activity was carried out via hydroxylation of the synthetic substrate p-nitrophenol to p-nitrocatechol. Crude microsomal preparation isolated from rat liver served as source for cytochrome P450 2E1. Under assay conditions guaranteeing a linear course of the reaction the cytochrome P450 2E1 was stimulated in the presence of a 10(-6) dilution of As2O3 corresponding to 0.915 microM final concentration compared with control. All other concentrations of As2O3 used inhibited the enzyme activity more or less drastically. Furthermore, we used this enzyme system to study the influence of arsenicum album (As2O3) and potassium cyanatum (KCN) in homeopathically prepared (i.e., by consecutive 1:10 steps) and conventional dilutions. We found significant differences between the effects caused by homeopathic potencies (D) and equally concentrated dilutions on catalytic activity of cytochrome P450 2E1. Such differing effects were observed in the case of arsenicum album (As2O3) between D4/10(-4) and D6/10(-6) and in the case of potassium cyanatum (KCN) between D6/10(-6) and D12/10(-12). When we used glutathione-S-transferases and uricase we also found different effects mediated by potencies and conventional dilutions. The results obtained suggest that these three enzyme systems are appropriate detection systems to hunt out differing effects of differently prepared dilutions of specific test substances. PMID- 9395662 TI - New developments in Cuban holistic medicine: a personal view. PMID- 9395663 TI - Familiarization with complementary medicine: report of a new course for primary care physicians. AB - A course entitled "Familiarization in Complementary Medicine" has been established at Exeter University. The syllabus of the course is presented, together with significant issues for debate that were raised by the attending primary care physicians. The delegates started with a positive but questioning attitude toward complementary medicine (CM) and acknowledge that they gained useful information, leading to increased confidence in discussing CM with patients. The course to a large extent met their needs and expectations. Benefits and draw-backs of integrating CM within general practice were explored. The main advantage of CM, apart from the potential intrinsic value of the techniques themselves, was identified as the time to establish a good therapeutic relationship with the patient. The particular concerns about CM that were expressed by the doctors included poor dialogue with CM practitioners, doubts about competence, and lack of readily identifiable and recognized qualifications. The risk of holding out unrealistic hope of cure was their greatest concern, however, especially if patients were thereby denied an effective orthodox treatment. The course was popular and will be repeated on a regular basis; similar courses for other health professionals including nurses are being planned. PMID- 9395664 TI - At least 80% of the population of most developing countries relies on traditional medicine as its primary source of health care. PMID- 9395665 TI - Global initiative for traditional health systems. PMID- 9395666 TI - Traditional medical practitioner in Vietnam. PMID- 9395667 TI - An aqueous extract of the leaves of Chromolaena odorata (formerly Eupatorium odoratum) (Eupolin) inhibits hydrated collagen lattice contraction by normal human dermal fibroblasts. AB - Chromolaena odorata (formerly Eupatorium odoratum) is used as a traditional medicine in Vietnam (Nghiem, 1992), where its Vietnamese common name is "co hoi." While it has been widely considered a weed by agriculturalists (Holm et al., 1991), the aqueous extract and the decoction from the leaves of this plant have been used throughout Vietnam for the treatment of soft tissue wounds, burn wounds, and skin infections. A number of clinical studies done by Vietnamese as well as foreign medical workers has demonstrated the efficacy of this extract on the wound-healing process. In this article, the effect of the Eupolin extract on hydrated collagen lattice contraction by human dermal fibroblasts, an in vitro model of wound contraction, is described. The significant inhibition of collagen gel contraction by Eupolin extract at 50 to 200 micrograms/ml is demonstrated in various concentrations of collagen. When the extract at 50 to 150 micrograms/ml was washed out of the lattices and replaced by fresh medium without Eupolin, the contraction of collagen by cells was resumed. The visualization of cells in the lattices by incubation in a tetrazolium salt for 2 h showed live cells at 50 to 150 micrograms/ml of extract. In contrast, all cells were killed in the higher extract doses of 300 or 400 micrograms/ml. These preliminary results showing the inhibitory effect of Eupolin extract on collagen contraction suggest that a clinical evaluation of its effect on wound contraction and scar quality should be made. This work illustrates that traditional remedies that are used by folk practitioners to improve healing can be examined in a scientific manner using in vitro wound-healing models. It could be that the synergistic properties of components of the natural extract contribute to the positive effects demonstrated on various wound-healing mechanisms. PMID- 9395668 TI - Effect of honey versus thyme on Rubella virus survival in vitro. AB - In this paper, we assess the antiviral properties of honey solutions and thyme extracts at varying concentrations. This was done by testing these solutions in vitro using monkey kidney cell cultures that were infected with the Rubella virus. Our results indicated that honey had good anti-Rubella activity, while thyme did not. These results may justify the continuing use of honey in traditional medicines from different ethnic communities worldwide and in some modern medications such as cough syrups. PMID- 9395669 TI - Conserving the medicinal plants of India: the need for a biocultural perspective. PMID- 9395670 TI - Ginkgo biloba L.: history, current status, and future prospects. AB - In this paper, we describe the status of the exploration and use of the Ginkgo biloba leaves in China. We emphasize the need for careful studies of the intra specific genetic diversity of Ginkgo biloba since genetic variation within this species may result in significant differences between the chemical and pharmacological properties of the different sub-species. It is therefore imperative that we catalog the intraspecific diversity of Ginkgo biloba L., and we stress that it is important to conserve this diversity since different subspecies may have different pharmacochemical properties resulting in differential medical usages. PMID- 9395671 TI - Research on medicinal plants and traditional medicine in Africa. PMID- 9395672 TI - Bumetha Rukararwe: integrating modern and traditional health care in southwest Uganda. AB - Rukararwe-Bumetha in southwest Uganda is a community-based program which combines the services of traditional medical practitioners with modern medical services in providing sustainable rural health care. In five years, approximately 50,000 patients have been seen. The clinic is staffed by a trained nurse and a pharmaceutical technician, and visited by doctors from nearby Mbarara University Medical School, all of whom work with traditional herbalists in diagnosing, standardizing dosage and consistency of medicines, and dispensing herbal mixtures, many of which are cultivated in Rukararwe's medicinal plant garden. Conservation of medicinal plants and education are central to the integrated health care services at Bumetha. Community health education emphasizes hygiene and the appropriate use of local medicinal resources. Traditional health practitioners care for the emotional and spiritual, as well as the physical, well being of their patients. PMID- 9395673 TI - The formulation of a health research agenda by and for indigenous peoples: contesting the Western scientific paradigm. AB - The paper addresses concerns related to the impact of Western science and technology upon indigenous communities in the face of current heightened globalization trends. Elements of the Western scientific paradigm that are antagonistic to indigenous ways of life and can result in detrimental cultural disruption are identified. In examining the Andean health paradigm, conceptual elements inherent to indigenous forms of knowledge generation are uncovered. The paper offers recommendations for the formulation of a research agenda and health promotion strategies aimed at maximizing the benefits of cultural interaction while minimizing potential damage to indigenous peoples. PMID- 9395674 TI - Physical activity. PMID- 9395675 TI - The genesis of physical culture in America. PMID- 9395676 TI - The antioxidant and antiatherogenic effects of MAK-4 in WHHL rabbits. AB - Oxidized low-density lipoprotein (ox-LDL) plays an important role in atherogenesis. Atheroma formation is reduced significantly in Watanabe heritable hyperlipidemic (WHHL) rabbits by antioxidants such as probucol and vitamin E. The herbal mixture Maharishi Amrit Kalash-4 (MAK-4) inhibits Cu+2 -induced LDL oxidation, and enzymatic- and nonenzymatic-induced microsomal lipid peroxidation. We tested the effect of MAK-4 on the development of atheroma in WHHL rabbits. Eleven rabbits were divided into two groups: controls (n = 5) and a group fed 6% (w/w) MAK-4 (n = 6). Blood was drawn for biochemical analysis every two months and at necropsy, six months after the special diet was started. The aortas were preserved in formalin. The percentage area of aortic arch covered with visible plaque in the MAK-4 group (22.5 +/- 4.2%, mean +/- SE) was significantly reduced (p < 0.01) compared to the control group (47.6 +/- 6.8%, mean +/- SE). The MAK-4 group showed a significant decrease (p < 0.05) in lipid peroxide, and a significant increase (p < 0.05) in glutathione peroxidase and resistance of LDL to endothelial cell-induced and cupric ion-catalyzed oxidation (4.5 h and 5 h lag phase, respectively, for the MAK-4 group; 0 h lag phase for both for the controls). These findings suggest MAK-4 reduces atheroma formation through its antioxidant activity. PMID- 9395677 TI - Physiological measures of right nostril breathing. AB - This study was conducted to assess the physiological effects of a yoga breathing practice that involves breathing exclusively through the right nostril. This practice is called surya anuloma viloma pranayama (SAV). Twelve volunteers (average age 27.2 years +/- 3.3 years, four males) were assessed before and after test sessions conducted on two consecutive days. On one day the test session involved practicing SAV pranayama for 45 minutes (SAV session). During the test period of the other day, subjects were asked to breathe normally for 45 minutes (NB session). For half the patients (randomly chosen) the SAV session was on the first day and the NB session on the next day. For the remaining six patients, the order of the two sessions was reversed. After the SAV session (but not after the NB) there was a significant (P < .05, paired t test) increase in oxygen consumption (17%) and in systolic blood pressure (mean increase 9.4 mm Hg) and a significant decrease in digit pulse volume (45.7%). The latter two changes are interpreted to be the result of increased cutaneous vasoconstriction. After both SAV and NB sessions, there was a significant decrease in skin resistance (two factor ANOVA, Tukey test). These findings show that SAV has a sympathetic stimulating effect. This technique and other variations of unilateral forced nostril breathing deserve further study regarding therapeutic merits in a wide range of disorders. PMID- 9395678 TI - Electrostimulation: addiction treatment for the coming millennium. AB - At a period of fundamental review of the health care system, it is timely to re assess one of medicine's most intractable problems--the treatment of addictions. The apparently insoluble dilemmas posed by the acute and chronic withdrawal syndromes underlie universally high drop-out and relapse rates. In a decade of HIV and AIDS infection, poly-substance addiction, potent street drugs, and ossified treatment strategies, it is urgent that policy formulators investigate seriously a flexible system of non-pharmacological transcranial electrostimulation treatment, based on its record of rapid, safe, and cost effective detoxification in several countries, as one innovative contribution to the challenges presented by addiction in the 1990s. This is a brief report of the introduction of NeuroElectric Therapy (NET) into Germany, describing the responses of the first 22 cases. The daily progress of a heroin addict and a methadone addict are detailed: both were treated as outpatients for 8 hours daily, for 7 and 10 days respectively. PMID- 9395679 TI - Wound healing and complementary therapies: a review. AB - A series of five innovative experiments conducted by Wirth et al. which examined the effect of various complementary healing interventions on the reepithelialization rate of full thickness human dermal wounds was assessed as to specific methodological and related factors. The treatment interventions utilized in the series included experimental derivatives of the Therapeutic Touch (TT), Reiki, LeShan, and Intercessory Prayer techniques. The results of the series indicated statistical significance for the initial two experiments and nonsignificance or reverse significance for the remaining three studies. This review article examines the methodological designs of the series of studies, along with the TT practitioners' phenomenologically based journal reports, to provide potential contributing correlative factors for the differential results obtained. These factors include: (1) methodological design restrictions, (2) a transference/inhibitory effect (3) the influence of experimental assistants, (4) healer visualization /imagery techniques, (5) variations in subject populations, and (6) a potential cancellation effect. While the placebo controlled double blind methodological designs used in the series were as stringent as those used in other fields of scientific inquiry, the overall results of the experiments were inconclusive in establishing the efficacy of the treatment interventions for accelerating the rate of reepithelialization of full thickness dermal wounds. PMID- 9395680 TI - The integrative hospital explored via acupuncture. AB - To integrate complementary and allopathic medicine within a single hospital requires close attention to models of health care delivery, economics, and medical philosophy. The practice of acupuncture can be used as a heuristic device to examine these issues and how solutions may apply to other complementary modalities in the creation of such a hospital facility. PMID- 9395681 TI - Methodological issues in complementary and alternative medicine research: a personal reflection on 10 years of debate in the United Kingdom. AB - There are certain immediate and obvious problems in using conventional research techniques for the evaluation of complementary and alternative medicine (CAM). These have led some to argue that alternative medicine requires alternative methodologies. The experience in the United Kingdom has been that existing methodologies can be adapted and do not have to be discarded wholesale. Conventional research has developed a set of powerful techniques for generating reliable knowledge and these have been used with considerable success in a variety of different settings. In retrospect, the call for new methodologies appears to have been a reaction against the perceived dominance of the randomized controlled trial (RCT). This stemmed from the misperception that the RCT inevitably involves features, such as placebo controls or double-blinding, which are not feasible in many CAM therapies. There was also a desire to ask questions about medicine, the answers to which, it was felt, could not be decided by the RCT. A variety of different research designs need to be used to answer the variety of questions important in CAM. However, research designs do not have to be reinvented: appropriate methodologies can normally be found in one or another of the diverse branches of medical research. Successful research has been conducted in CAM using conventional research techniques and this refutes the claims that such techniques are inappropriate for CAM. Solving methodological problems in CAM is a matter of following simple guidelines, not the creation of complex and esoteric research designs. PMID- 9395682 TI - The traditional use of maggots in wound healing, and the development of larva therapy (biosurgery) in modern medicine. PMID- 9395683 TI - Post modern medicine. AB - Although there is general agreement that our current approach to health and healing is undergoing substantial change, there has been a lack of critical discussion regarding the extent, character, and direction of that change. It is important for us to articulate the values that we would like expressed in a reconfigured approach to health and healing, and to ensure that our efforts to initiate change are aligned with these values. An overview of western history suggests that the post-modern world view is characterized by four essential values: multidimensional realism, intentionality, holism, and personal authenticity. Our current efforts to change the direction of health care have failed to produce fundamental change. This has resulted from the compelling influence of existing perspectives and the promotion of parochial professional and institutional interests. To assure fundamental change, values that are consistent with a post-modern world must be articulated, fostered, and embedded into innovative programs. PMID- 9395684 TI - Challenges for the future: report on the First National Conference on Medical and Nursing Education in Complementary Therapies. PMID- 9395685 TI - Report of the National Workshop on Research Methodologies for Unconventional Therapies, sponsored by the Canadian Breast Cancer Research Initiative (CBCRI), held October 4-6, 1996, at the Delta Pacific Resort and Conference Center in Richmond, B.C. PMID- 9395686 TI - Report from the Cochrane Collaboration 4th International Colloquium, held in Adelaide, Australia, October 20-24, 1996. PMID- 9395687 TI - One man's meat is another man's poison: the challenge of psychic/intuitive diagnosis to the diagnostic paradigm of orthodox medical science. PMID- 9395688 TI - Phrenologist at work, 1856. PMID- 9395689 TI - A comparative survey of leguminous plants as sources of the isoflavones, genistein and daidzein: implications for human nutrition and health. AB - Over 80 taxa of mostly agriculturally important legumes were surveyed as sources of the metabolites, genistein and daidzein. Remarkably high concentrations (over 2 g.kg-1 dry weight) of the anticancer metabolite, genistein, were found in the leaves of Psoralea corylifolia (Indian bread root). All other legumes, with the exception of fermented soybean miso, had genistein levels < 400 mg.kg-1 dry weight. Concentrations of over 1 g.kg-1 dry weight and 0.95 g.kg-1 dry weight of the anticancer metabolite, daidzein, were found in the stems of the fava bean (Vicia faba) and roots of kudzu vine (Pueraria lobata), respectively. From this survey, our results indicate that the legumes, lupine (Lupinus spp.), fava bean, (Vicia faba), soybeans (Glycine max), kudzu (Pueraria lobata), and psoralea (Psoralea corylifolia), are excellent food sources for both genistein and daidzein. Miso, a fermented soybean product, is also a rich source of both isoflavones. PMID- 9395690 TI - Microbiological screening of Indian medicinal plants with special reference to enteropathogens. AB - The World Health Organisation (WHO) has recommended that all member states actively promote native medicines in their country. Ten Indian medicinal plants were screened for antibacterial activity specific to enteropathogens. Diffusion and dilution methods were used to measure the antibacterial activity. Allium sativum, Camellia sinensis, and Chamaesyce hirta showed higher activity when compared to the rest. They had a minimum bactericidal concentration (MBC) of < 100 micrograms/ml and gave inhibition zones of more than 2 cm. Among the pathogens studied, Vibrio cholerae and Shigella flexneri were found to be highly susceptible to the plant extracts. PMID- 9395691 TI - Double-blind study of pulsing magnetic field effects on multiple sclerosis. AB - We performed a double-blind study to measure the clinical and subclinical effects of an alternative medicine magnetic device on disease activity in multiple sclerosis (MS). The MS patients were exposed to a magnetic pulsing device (Enermed) where the frequency of the magnetic pulse was in the 4-13 Hz range (50 100 milliGauss). A total of 30 MS patients wore the device on preselected sites between 10 and 24 hours a day for 2 months. Half of the patients (15) randomly received an Enermed device that was magnetically inactive and the other half received an active device. Each MS patient received a set of tests to evaluate MS disease status before and after wearing the Enermed device. The tests included (1) a clinical rating (Kurtzke, EDSS), (2) patient-reported performance scales, and (3) quantitative electroencephalography (QEEG) during a language task. Although there was no significant change between pretreatment and posttreatment in the EDSS scale, there was a significant improvement in the performance scale (PS) combined rating for bladder control, cognitive function, fatigue level, mobility, spasticity, and vision (active group -3.83 +/- 1.08, p < 0.005; placebo group -0.17 +/- 1.07, change in PS scale). There was also a significant change between pretreatment and posttreatment in alpha EEG magnitude during the language task recorded at various electrode sites on the left side. In this double-blind, placebo-controlled study, we have demonstrated a statistically significant effect of the Enermed magnetic pulsing device on patient performance scales and on alpha EEG magnitude during a language task. PMID- 9395692 TI - Characteristics and complaints of patients seeking therapy at a hospital-based alternative medicine clinic. AB - BACKGROUND: Patient use of complementary and alternative medicine (CAM) is on the rise. With millions of Americans using CAM, it has become imperative from the public health point of view to undertake a coordinated research effort that will thoroughly evaluate the role and effectiveness of CAM modalities. METHODS: We developed a prospective data-collection system to capture presenting complaints, patient health histories, and demographic information on the patients of a hospital-based alternative medicine clinic. RESULTS: Of 760 patients in the present cohort, 248 different complaints or complaint combinations were entered. Of 16 major categories, the largest was musculoskeletal, followed by the addictions, psychiatric, and nonspecific categories. Slightly more than one in five patients requested treatment on the recommendation of their physician. Over two-thirds of patients in this study group were female. CONCLUSIONS: Given the growing interest and use of complementary and alternative therapies, a system such as described can demonstrate the types of patients presenting for treatment, a more detailed picture of their complaints, and, over time, measurable outcomes. PMID- 9395693 TI - An experimental test of psychic diagnosis of disease. AB - The authors, one a medical anthropologist and the other a medical doctor, investigated the claims of three acquaintances who include psychic diagnosis in the services they provide to clients. An experiment was designed that allowed each of the three psychics to diagnose five volunteers, patients of S. K. H. Aung, one at a time while hidden behind a screen. The results of the psychic diagnoses were then compared with the medical records of the patients. Results indicate some correspondence between the psychic diagnoses and the medical records, but the correspondence was not sufficiently impressive to warrant considering psychic diagnosis as a useful alternative method for diagnosing disease. It would appear that patients relying solely on psychic diagnosis as the basis for therapy are at risk of serious medical problems going undetected. Future research, with a larger sample of patients and psychic diagnosticians, is required to substantiate this conclusion. PMID- 9395694 TI - A review of research on acupuncture for the treatment of lumbar disk protrusions and associated neurological symptomatology. AB - The association between acupuncture (AP) and pain relief is so strong that it has tended to obscure any other potentially significant clinical results. This review indicates the wealth of data from around the world on various aspects of AP treatment for low back syndromes related to lumbar intervertebral disk prolapse (PID). Although plentiful, the research is variable in quality, especially with respect to design, consistency, and follow-up. Even so, the large number of patients who appear to have been treated successfully (i.e., given a high degree of symptomatic relief) supports a potential role for AP. This is further supported by studies on patients who had previously had unsuccessful treatment with conservative methods. The role envisaged for AP, in cases of lumbar PID and sciatica, is at least of a supplementary therapy capable of reducing the requirement of more invasive forms of treatment. No such role is envisaged in cases of cauda equina compression where surgery must remain the treatment of choice. AP should be explored more fully, using appropriate designs, so that this discipline may achieve its full therapeutic potential in the West. PMID- 9395696 TI - Redefining medicine for the 21st century: reflections on values and principles: a personal view. PMID- 9395695 TI - Two cases of hepatitis C treated with herbs and supplements. AB - The treatment of two cases of hepatitis C using herbal medicine and nutritional supplements is presented. The selection of medicinals was based upon both biomedical findings and traditional Chinese medical diagnosis. The text describes the course of each patient's illness documented both subjectively and objectively using blood values and traditional Chinese medicine analysis as parameters. Explanation and/or citations are given for each medicinal used. Both patients improved during the course of treatment; subjective signs and symptoms (especially fatigue) as well as liver enzyme levels demonstrated improvement. PMID- 9395697 TI - Maryland massage therapy bill passes after 10 years. PMID- 9395698 TI - Dramatic increase in the interest in, and, use of, alternative and complementary medicine. PMID- 9395699 TI - Hydrotherapy. PMID- 9395700 TI - Multisite electromyographic analysis of therapeutic touch and qigong therapy. AB - The influence of complementary healing treatment on paraspinal electromagnetic activity at specific neuromuscular sites was examined in an exploratory pilot study that used a multisite surface electromyographic (sEMG) assessment procedure. The study was a replication and extension of previous research that indicated that complementary healing had a significant effect in normalizing the activity of the "end organ" for the central nervous system (CNS). Multisite sEMG electrodes were placed on the frontalis, cervical (C4), thoracic (T6), and lumbosacral (L3) paraspinals of 44 subjects who were divided into three groups: (1) students/patients of a Qigong practitioner (n = 16); (2) students/patients of a therapeutic touch (TT) practitioner (n = 14); and (3) nonbelievers in complementary healing (n = 14). A traditional ABAC experimental design was used with each subject evaluated for one 20-minute session that included four 5-minute segments. The purpose of this study was to measure the variable energizing effect of Qigong therapy along with the anecdotally and experimentally established relaxation effect of TT therapy relative to patient belief and expectancy. Treatment sessions consisted of Qigong and a modified form of TT intervention for all three groups. Due to the double-blind nature of the study, however, group 1 subjects were aware of only the Qigong intervention; group 2 subjects were aware of only the TT intervention, and group 3 subjects were informed that the study was designed to assess the neuromuscular activity of individuals in a seated position. The results indicated a statistically significant rise in electromagnetic activity for group 1 during the Qigong intervention segment (p < .024). Group 2 demonstrated a modest although overall nonsignificant decrease in multisite sEMG levels for both treatment protocols, whereas group 3 exhibited relatively consistent neuromuscular activity for both control and treatment segments. The results of this study are considered preliminary in nature, however, due to the potential influence of several confounds including psychophysiological factors, established behavior patterns, and the possibility for information transfer due to sensory cues. PMID- 9395701 TI - Use of acupuncture by American physicians. AB - OBJECTIVES: Little is known about the characteristics of American physicians who currently practice acupuncture. We asked: (1) Do the demographics of physicians practicing acupuncture differ from the general physician population? (2) Do these physicians use or endorse other alternative therapies? (3) For which conditions is acupuncture most commonly used? (4) For which conditions is acupuncture perceived to be most efficacious? DESIGN: Mailed survey of physicians who incorporate acupuncture into their practice. PARTICIPANTS: Membership of the American Academy of Medical Acupuncture (AAMA). OUTCOME MEASURES: Demographic information regarding physicians and practice characteristics; specific illnesses treated, and perceived efficacy; use of other complementary modalities; personal reasons for practicing acupuncture. RESULTS: Compared with national data, respondents were more likely to be nonspecialists, in private practice, and age 35 to 54. There was an equal proportion of men and women. Most had been doing acupuncture for < 5 years; most use it on < 25% of their patients. Endorsement or use of other complementary methods (spinal manipulation, herbal medicine, supplements, homeopathy) was common. Acupuncture was more commonly used for pain conditions than general medical problems or addiction management. Reasons for use included: efficacy of the technique, an alternative in cases of inadequacy of standard medical approach, and a multidimensional approach to health care. CONCLUSIONS: Physicians surveyed in this study who incorporate acupuncture into their practice do so mainly to treat pain problems. They are more likely to be in the 35 to 54 age group, nonspecialists, and in private practice when compared with national averages. These physicians are also more likely to use or endorse other complementary modalities. PMID- 9395702 TI - Who seeks alternative health care? A profile of the users of five modes of treatment. AB - This article compares the social and health characteristics of patients of five kinds of practitioners: family physicians (used as a baseline group); chiropractors; acupuncturist/traditional Chinese medicine doctors; naturopaths; and Reiki practitioners. The data were gathered in a large Canadian city during the period 1994 to 1995. Face-to-face interviews were conducted with 300 patients (60 from each type of treatment group). While the most striking social and health differences occur between patients of family physicians and the patients of alternative practitioners, significant differences are also evident between the different groups of alternative patients. Reiki patients, for example, have a higher level of education and are more likely to be in managerial or professional positions than other alternative patients. The profiles presented here indicate that users of alternative care should not be regarded as a homogeneous population. The findings also show that almost all alternative patients also consult family physicians. The pattern revealed is one of multiple use: patients choose the kind of practitioner they believe can best help their particular problem. PMID- 9395703 TI - Researching complementary therapies: a Delphi study to identify the views of complementary and orthodox practitioners. AB - Twenty five participants were invited to a workshop looking at the role of research and development in complementary therapies. These participants took part in a modified Delphi study to ascertain a consensus from their broad spectrum of views and see how those views changed over time. Opinions were sought about respondents' perceptions of complementary therapies, the type and scope of research needed, and how such research should be coordinated. The workshop, arranged by the Regional Health Authority, included orthodox and complementary practitioners, purchasers, providers and academics. Questionnaires were administered at the beginning and end of the workshop and 6 months later. The group was small and potentially positively biased, having both pre-existing knowledge and interest in complementary therapies. In consensus, the more mainstream complementary therapies of osteopathy, acupuncture, and homeopathy were perceived to be the most beneficial to patients, and therefore, most likely to be recommended as treatments. These were also felt to be the priority areas for research. For some unknown reason homeopathy had lost its popularity in ranking 6 months later. The respondents agreed that research into complementary therapies was needed and ideally should be collaborative between practitioners, professional bodies, and academics. The workshop appeared to produce some changes in knowledge that were not always maintained over time. It was, however, formative in the decision of the Regional Health Authority to allocate research funding into complementary therapies. PMID- 9395704 TI - Effect of the combination of Aloe vera, nitroglycerin, and L-NAME on wound healing in the rat excisional model. AB - PURPOSE: Many systemic and topical therapeutic agents such as growth hormone, platelet-derived growth factor (PDGF), fibroblast growth factor (FGF), epidermal growth factor (EGF), and insulin-like growth factor (IGF) have been used as vulnerary agents. However, the role of nitric oxide (NO) as a wound-healing stimulant has been received with mixed reviews. NO is a potent vasodilator that is thought to be an endothelium-dependent relaxing factor, and a regulator of blood pressure and regional blood flow. It affects vascular smooth muscle proliferation and inhibits platelet aggregation and leukocyte adhesion. Therefore we compared the effects of several topical substances that have similar or reverse properties. METHODS: Using the excisional rat wound model, we evaluated the topical effects of Dermaide Aloe (D-Aloe, Dermaide Research Corp, Palos Heights, IL), nitroglycerin, Aquaphor (Beuersdorf, Inc., Norwalk, CT) alone, with D-Aloe with nitroglycerin, 2%, and L-NAME (NO inhibitor) with Aquaphor, and L NAME with Aquaphor and D-Aloe for a 21-day period. All wounds were measured by planimetry at 1, 7, 10, 13, 16, 18, and 21 days. RESULTS: At day 1, all wounds had an average wound size of 2.27 cm2 (SD +/- 0.372) with no significant difference in wound size among the groups. Topically applied D-Aloe appeared to promote wound healing faster than the remaining other topicals (p < .05, Student Newman-Keuls and Dunn's Method) over the study period. However, topicals combined with D-Aloe, the vehicle Aquaphor, and L-NAME improved the wound healing process when compared with nitroglycerin alone (p < .05). CONCLUSIONS: D-Aloe appears to have a wound-healing advancement factor that can reverse the effects of petrolatum- and nitroglycerin-based products as observed in the remaining groups when compared with nitroglycerin alone. It appears that D-Aloe's effect of preventing dermal ischemia by reversing the effects of thromboxane synthetase (TxA2) may act synergistically with NO or could be an oxygen radical scavenger. PMID- 9395705 TI - Successful treatment of herpetic infections by autohemotherapy. AB - Herpes zoster (shingles) affects a significant number of individuals over age 50. To date, no satisfactory treatment has been available. The clinician author (JHO) witnessed a dramatic response of a shingles patient to autohemotherapy: the pain was completely relieved and lesions gone within 5 days with no recurrence of either. Treatment of other herpetic patients then began with autohemotherapy. Twenty-five patients with herpes were given an autologous blood transfer of 10 mL of blood from the antecubital vein into the gluteal bundle and followed for clinical signs. A 100% favorable response occurred in 20 patients who received autohemotherapy within 7 weeks of the onset of clinical signs and 1 other who received autohemotherapy at a 9-week interval. No untoward signs or symptoms of the treatment occurred. Autohemotherapy has been demonstrated to be effective in elimination of clinical sequelae in these cases of herpes infections and these results justify further rigorous clinical investigation. PMID- 9395706 TI - When conventional treatment is not enough: a case of migraine without aura responding to homeopathy. AB - A case of migraine without aura is reported, unresponsive to five years of treatment with very well indicated conventional therapies, which are listed in detail. Consultation with a homeopathic physician, who also has extensive experience in diagnosis and treatment of headache disorders, leads to the prescription of a single homeopathic remedy which was absolutely effective for the condition. This case is offered as an open, admittedly retrospective study, comparing the best of conventional migraine therapy with appropriate homeopathic therapy in the same patient. PMID- 9395707 TI - Diarrhea and human immunodeficiency virus: Western and Eastern perspectives [corrected]. AB - Sixty percent of patients with human immunodeficiency virus (HIV) in the United States and 90% throughout the world will have diarrhea at some point in their illness. This article provides an introductory exploration and discussion of Western and Eastern perspectives of chronic diarrhea in patients with HIV. Western etiologies and treatment approaches, as well as Eastern views from traditional Chinese medicine pathogenesis and treatment principles involving acupuncture and moxibustion are presented. Whereas their interpretations of the causes of diarrhea are different, both the East and West have something to offer patients with this distressing symptom. Further exploration and clinical research is needed in this area. PMID- 9395709 TI - Segment therapy: the effects of ultrasound and benzocaine spray in the treatment of contractures and spasticity. AB - According to Krusen's Handbook of Physical Medicine and Rehabilitation, topical application of ultrasound exerts only an indirect and adjunctive effect as deep heat treatment. In support of this idea the extensibility of an isolated tendon in a heated water bath is described. The significance of this experiment as a biological phenomenon has never been critically examined. In this article the existence of a segmental reflex is proposed. Such a segmental paradigm is in keeping with the segmental organization and evolution of the locomotor system. It is further suggested that ultrasound and related therapy, if applied segmentally, may have a curative effect. PMID- 9395708 TI - HIV-related diarrhea: urgent need for a reasoned holistic response. PMID- 9395710 TI - Directory of Databases for research into alternative and complementary medicine. AB - This Directory of Databases with significant holdings of primarily bibliographic references to complementary and alternative medicine research resources has been compiled to facilitate access to the widely scattered data and literature. The Directory is directly accessible from the Web site of the Richard & Hinda Rosenthal Center for Complementary and Alternative Medicine at Columbia University's CPMCNet (http://cpmcnet.columbia.edu/dept/rosenthal/). General selection criteria and a brief description of content, access, or contact details are given for each of the 56 databases. Thirty-five of the databases are available online over the Internet and of these, 17 are freely, publicly available. There are 13 search services and a further 8 databases available in a variety of formats. PMID- 9395711 TI - The Cochrane Collaboration and evidence-based complementary medicine. PMID- 9395712 TI - Follistatin and its role as an activin-binding protein. AB - Follistatin (FS), a specific binding protein for activin, neutralizes the diverse actions of activin by forming an inactive complex with activin. FS is a monomer derived from two polypeptide core sequences of 288 (FS-288) and 315 (FS-315) amino acids originated from alternatively spliced mRNA. We purified six molecular forms of FS from porcine ovaries. Their structural differences were caused by truncation of the COOH-terminal region and/or the presence of carbohydrate chains, resulting in the formation of FS-288, FS-315 and FS composed of 303 amino acids (FS-303) in various forms of glycosylation on the two potential Asn-linked glycosylation sites. All six molecular species have almost the same activin binding activity (Kd = 540-680 pM). By contrast, the COOH-terminal truncated form, FS-288, showed much higher affinity for heparan sulfate proteoglycans of the cell surface than FS-303, whereas the intact form of FS, FS-315, had no affinity. Furthermore, FS-288 more effectively blocked the suppression of follicle-stimulating hormone (FSH) secretion from rat pituitary cells by activin. This implies that activin binds to the cell surface through FS-288 which adheres to the cell surface. To clarify the physiological role of cell-associated FS, we then investigated the binding of activin to cell-associated FS and the fate of cell surface-bound activin and FS using primary cultured rat pituitary and ovarian granuloma cells. When the cells were incubated with 125I-activin A in the presence of FS-288 or 315, the binding of activin A to the cell surface was promoted much more markedly by FS-288 than by FS-315. The amounts of radioactivity recovered in trichloroacetic acid-soluble fractions (degraded activin) from the incubation medium were greatly increased by the addition of FS 288. This increase was abolished by heparan sulfate, monensin (an endocytosis inhibitor), chloroquine (a lysosome function inhibitor) and several lysosomal enzyme inhibitors. These results suggest that cell-associated FS-288 accelerates the internalization of activin into the cells, leading to its degradation by lysosomal enzymes, and that cell surface-associated FS therefore plays a role in the clearance system of activin. PMID- 9395713 TI - Tumor suppressor genes in human lung cancer. AB - Lung cancer is the most common cause of cancer death in Japanese males, the incidence having increased markedly in recent years. Carcinogen exposure such as to tobacco-smoke and air pollution are associated with the probability of developing lung cancer. Aquired somatic mutations play an important role in the pathogenesis of environmentally induced lung cancers. Cytogenetic and molecular analysis of lung tumors has made it possible to examine this hypothesis and to search for candidate genes that may be targeted by chronic exposure to these carcinogens. Early studies implicate several distinct chromosomal loci (3p, 9p, 13q, 17p, and others) and suggest sequential genetic events occur during the initiation and progression of lung carcinogenesis. Several suppressor genes including Rb (13q), P53 (17p), and P16 (9p) have been identified and cloned at these chromosomal loci. The identification of putative tumor suppressor gene at chromosome 3p is still under work. Understanding the interaction of P53, RB, cyclins, and protein kinase inhibitors including P16 will be essential to the development of the next generation of diagnostic and therapeutic studies for lung cancer. PMID- 9395714 TI - Peripheral blood stem cell transplantation; an update. AB - Patients with a number of different malignancies have been treated with high-dose chemotherapy and peripheral blood stem cell transplantation (PBSCT). PBSC already replaced bone marrow as the source of autologous hematopoietic progenitor support. This is due to ease of collection, rapid engraftment and less possibility of tumor cell contamination in the graft. Furthermore, allogeneic transplantation of granulocyte colony-stimulating factor (G-CSF) mobilized PBSC is now being increasingly performed. Recent advance of clinical PBSCT and new strategies are stressed in this review. New strategies include CD 34+cell purification, ex vivo expansion of PBSC and PBSC as a target cell for gene therapy. Major future advance may occur better understanding of the mechanism of mobilization and the biology of PBSC. PMID- 9395715 TI - Cathepsin B-inhibitor promotes the development of Th1 type protective T cells in mice infected with Leishmania major. AB - BALB/c mice are genetically susceptible to infection with Leishmania major (L major). When such mice infected with L. major were treated with specific inhibitors of cathepsin B, a lysosomal cysteine protease that digests exogenous antigenic proteins, the mice acquired resistance against L. major infection. T cells from these mice produced large amounts of IFN-gamma and low amounts of IL-4 as compared with those of untreated BALB/c mice. In addition, the mice treated with cathepsin B inhibitor produced a high titer of IgG2a specific antibodies and only low titers of IgG1 and IgE antibodies. This type of response is in contrast with the high specific IgG1 or IgE antibody responses which are the usual antibody responses in BALB/c mice infected with L. major. These findings indicate that cathepsin B may be critically involved in processing antigens of L. major to promote exclusively the development of Th2 type CD 4+T cell responses. PMID- 9395716 TI - Visual evoked potential and electroencephalogram of healthy females during the menstrual cycle. AB - Flash visual evoked potential (VEP) and electroencephalogram (EEG) changes during the menstrual cycle were studied using healthy females having regular menstruation, with 21 at the follicular phase (FP) and 23 at the luteal phase (LP). The following results were obtained. (1) The waveforms of Group Mean VEPs of both groups had approximately similar triphasic contours, consisting of 16 components of P 1-N 8 up to 500 msec of latency. (2) Latencies tended to be longer in LP. (3) Interpeak amplitudes tended to be larger in LP, and one VEP interpeak amplitude (P 5-N 7) of long latency component was significantly larger at LP after eliminating the effect of body height by ANCOVA for 2 CH. (4) Quantitative analysis of EEGs between FP and LP resulted in a tendency for increased alpha, and decreased beta power % at LP. Since estrogen increases the VEP amplitude, and decreases the VEP latency and the alpha activity of EEGs, the large VEP amplitude, the tendency for prolonged VEP latency, and the tendency for increased alpha power % at LP observed in this study indicate that the VEP amplitude at LP reflects the effect of estrogen, and that the VEP latency and EEGs at LP reflect the effect of progesterone. PMID- 9395717 TI - Bile-induced DNA strand breaks and biochemical analysis of bile acids in an experimental model of anomalous arrangement of the pancreaticobiliary ducts. AB - A canine experimental model for the anomalous arrangement of the pancreaticobiliary ducts (APBD) was made to investigate the effects of bile acids on carcinogenesis. Seven adult mongrel dogs underwent dorsal pancreatico cholecystostomy to serve as a functional model for APBD, and six dogs underwent the same procedure with the pancreatic duct ligated as a control group. Bile from the gallbladder was taken 14 months after surgery for bile acid analysis by HPLC. DNA strand breaks in HeLa cells induced by the bile were also investigated in situ by nick translation method. As a result, the fraction of cholic acid tended to be lower, and that of deoxycholic acid slightly higher in APBD-dogs (N.S.). The ursodeoxycholic acid percentage in APBD-dogs significantly decreased compared with that in the control and normal dogs (p < 0.05). Extremely high frequency of DNA strand breaks was shown in only two out of seven APBD-dogs. In those two dogs, the cholic acid percentage decreased and that of deoxycholic acid increased extremely. These findings suggest that the alteration of the bile composition in APBD caused frequent DNA strand breaks and repair which might lead to gene mutation and biliary tract carcinoma. PMID- 9395718 TI - Interleukin-8 in bronchoalveolar lavage fluid of patients with diffuse panbronchiolitis or idiopathic pulmonary fibrosis. AB - This study was designed to clarify the contribution of IL-8 as a specific neutrophil chemotactic factor in the human respiratory tract in various pulmonary diseases. The neutrophil chemotactic activity (NCA), neutrophil counts and IL-8 concentration in the bronchoalveolar lavage fluid (BALF) obtained from normal volunteers (NV), control patients (CP), patients with diffuse panbronchiolitis (DPB) and patients with idiopathic pulmonary fibrosis (IPF) were examined. Neutrophil counts, NCA and IL-8 concentration in BALF obtained from patients with DPB or IPF was significantly higher than that from NV or CP. The IL-8 concentration correlated with neutrophil count and also correlated with NCA in BALF from patients with IPF, whereas there was no correlation between these factors in BALF from DPB. These results suggest that the contribution of IL-8 to neutrophil accumulation of the lower respiratory tract is different between IPF and DPB. PMID- 9395719 TI - Thrombin stimulates platelet-derived growth factor release by alveolar macrophages in rats--significance in bleomycin-induced pulmonary fibrosis. AB - Thrombin is a multifunctional enzyme generated at sites of vascular injury, and is known to be increased in the lungs in some types of fibrotic lung disease. In this study, to determine whether thrombin is associated with fibroblast growth and pulmonary fibrosis in these disorders, we examined whether a growth factor for fibroblasts (platelet-derived growth factor, PDGF) was released by thrombin stimulated alveolar macrophages (AM). The culture supernatants of rat AM stimulated with 1 or 10 U/ml of thrombin showed a significant increase in fibroblast growth-stimulating activity (FGA). Pretreatment of the AM supernatant with anti-PDGF-AA antibody significantly decreased the FGA, but pretreatment with anti-PDGF-BB antibody did not. The supernatants of AM stimulated with thrombin also increased the growth of fibroblasts from the lungs of rats with bleomycin induced lung injury. These results indicate that thrombin stimulates AM to release PDGF-AA, which is responsible, at least in part, for fibroblast growth and the development of pulmonary fibrosis in some types of fibrotic lung disease. PMID- 9395720 TI - Comparison of energy metabolism in insulin-dependent and non-insulin-dependent diabetes mellitus. AB - To compare the metabolic consequences of insulin-dependent diabetes mellitus (IDDM) and non-insulin-dependent diabetes mellitus (NIDDM), glycemic control and energy metabolism were evaluated in 18 children displaying IDDM and 19 NIDDM adult patients. With rising concentrations of fasting blood glucose (FBG), hemoglobin A1C and free fatty acid, the percentage of the ratio of resting energy expenditure (REE) to predicted REE expressed as %REE increased and the respiratory quotient (RQ) decreased. The linear regression between RQ and FBG showed the same gradient in IDDM and NIDDM although the RQ in IDDM was always 0.07 lower than that in NIDDM given various FBG concentrations. Those patients whose RQ values were less than 0.7, indicating ketone body production, included 8 (44%) IDDM and 2 (11%) NIDDM patients. These results may explain the relatively greater manifestation of ketoacidosis in IDDM. PMID- 9395721 TI - Effect of K+ channel openers on K+ channel in cultured human dermal papilla cells. AB - Minoxidil sulfate and pinacidil, well-known activators of the ATP-sensitive K+ (KATP) channel, induce hair growth in clinical studies. The opening of K+ channels is thought to be an important mechanism in the regulation of hair follicles. In the present study, we used the patch clamp technique to characterize the K+ channels and tested the effect of K+ channel openers on K+ channels in cultured human dermal papilla cells. In dermal papilla cells, the Ca(2+)-activated K+ (KCa) channel with large conductance (179.3 +/- 13.1 pS in symmetrical 150 mM K+ solutions, n = 9) was dominant and we could not observe KATP channels in cell-attached and inside-out patches. In addition, minoxidil and pinacidil failed to activate KATP or KCa channels. In inside-out membrane patches, the channel was blocked by 10 mM tetraethylammonium ion, 2 mM 4 aminopyridine to the cytosolic face of the membrane or by lowering Ca2+ using 10 mM EGTA, but not by glibenclamide. In the cell-attached patch configurations, extracellular application of 1 mM sodium nitroprusside, a nitrovasodilator, activated the KCa channel. Methylene blue (2 mM) inhibited channel activation by sodium nitroprusside. Extracellular application of 20 mM dibutyryl cGMP activated the KCa channel, suggesting that channel activation is mediated by cGMP. Nitrovasodilators, which have no effect on hair growth, now appear to activate KCa channels in dermal papilla cells. These results suggest that increased K+ permeability itself in dermal papilla cells may not be sufficient for promotion of hair growth. PMID- 9395722 TI - Effect of evodiamine on catecholamine secretion from bovine adrenal medulla. AB - The effect of evodiamine on catecholamine secretion from bovine adrenal medulla was investigated. Evodiamine, a bioactive component isolated from dry unripened fruit of Evodia rutaecarpa Bentham, was found to stimulate the secretion of catecholamine from perfused bovine adrenal medulla at a concentration of 10 microM and its effect persisted for at least 30 min. This stimulatory effect of evodiamine was abolished by omission of Ca2+ from the perfusion fluid. Evodiamine (0.1-10 microM) markedly enhanced the secretion of catecholamine from the adrenal medulla induced by acetylcholine (100 microM or high K+(56 mM). The secretion of catecholamine was promptly enhanced by acetylcholine or high K+, but returned to the control level on treatment for 20 min. However, when evodiamine was added to the perfusion fluid after acetylcholine or high K+ stimulation for 10 min, the secretion of catecholamine again increased greatly. These results indicate that evodiamine not only stimulated the secretion of catecholamine from bovine adrenal medulla but also reversed insensitivity of these cells to acetylcholine or high K+ stimulation. PMID- 9395723 TI - Estimation of free calcium levels after thyroidectomy. AB - Total calcium is routinely measured after thyroidectomy in a clinical setting, while the measurement or calculation of the free calcium level is not generally performed. We reviewed total and free calcium levels in patients who underwent lobectomy (n = 15), subtotal thyroidectomy (n = 15) and total thyroidectomy (n = 15). Postoperative total calcium levels decreased significantly in comparison to preoperative levels in all thyroidectomies (p < 0.01), and this fall was significantly related to the extent of surgery (p < 0.01). In contrast, there was no significant difference between preoperative and postoperative free calcium levels in patients undergoing lobectomy, although we found a decrease in free calcium levels after both subtotal and total thyroidectomy. Total protein levels decreased regardless of the type of operation. Serum total calcium levels were thought to be altered by serum protein levels through the change of protein-bound calcium levels. When examined for free calcium levels, some patients were administered unnecessary calcium supplementation because hypocalcemia had been judged from the total calcium level. Since the wrong diagnosis may be given with regard to hypoparathyroidism by measurement of total calcium levels alone, we propose that free calcium levels should be routinely measured or calculated after thyroidectomy. PMID- 9395724 TI - Age-related increase of autoantibodies to interleukin 1 alpha in healthy Japanese blood donors. AB - Although autoantibodies to interleukin-1 alpha (IL-1 alpha autoantibodies) are known to be present in sera of apparently healthy humans, their frequency of occurrence and significance are unclear. To determine the prevalence of detectable IL-1 alpha autoantibodies in normal human blood, we screened the plasma of blood donors (6290 subjects: 3977 men and 2313 women, ages 16 to 64 yr) by a radioimmununoassay which we developed using a method that could detect over 5 ng/ml. Moreover, we investigated immunoglobulin class of IL-1 alpha autoantibodies and also their function. IL-1 alpha autoantibodies were detected in 14.6% of the 6290 donors. Their frequency was higher in males than females (16.6% vs. 11.2%, p < 0.01) and increased with age in both sexes. The proportion of subjects with a high IL-1 alpha autoantibodies titers also increased with age. We showed that IL-1 alpha autoantibodies were of the IgG class and that they had neutralizing function to IL-1 alpha by receptor assay. Neutralizing activity was only shown in plasma with concentration of IL-1 alpha autoantibodies, the level of which was over 1000 ng/ml. The affinity of the IL-1 alpha autoantibodies in plasma was between 2.1 x 10(-10) and 1.2 x 10(-9) M (mean 6.4 x 10(-10)M). Our results provide a basis for comparison with IL-1 alpha autoantibodies prevalence between healthy states and disease states, and suggest that IL-1 alpha autoantibodies may play a significant role in modulating the effects of excessive IL-1 alpha at local site or in systemic regions. PMID- 9395725 TI - Two siblings with vitamin B6-nonresponsive cystathionine beta-synthase deficiency and differing blood methionine levels during the neonatal period. AB - We present two siblings with vitamin B6-nonresponsive homocystinuria due to a deficiency of cystathionine beta-synthase who had different levels of methionine in the blood during the neonatal period, even though they had the same genetic defect. One of them was missed in the screening of newborns for homocystinuria. Special care should be taken in screening neonates for homocystinuria using the blood level of methionine. PMID- 9395726 TI - Immunological functions of adult T cell leukemia cells of a patient complicated with synchronous double primary gynecologic cancer. AB - A patient with triple malignancies is reported, who presented cervical cancer, vulvar cancer and adult T cell leukemia (ATL). ATL was diagnosed as a smouldering type, because antibody to human T cell leukemia virus associated antigen (ATLA) was positive with a titer of 1:160. Although her malignant cells had an OKT 4+8 3+Tac+ phenotype, the cells did not display helper T cell functions. Namely they showed no response to Phytohemagglutinin (PHA) and Interleukin 2 (IL-2) and suppressed the PWM driven IgG synthesis of B cells obtained from healthy donor. They did not produce IL-2 by stimulation with PHA and phorbol myristate acetate (PMA). Furthermore, these ATL cells were producing IL-2 inhibitor like factors. As synchronous triple malignancies are extremely rare, two gynecologic cancers seem to ascribe to the suppressing state of the immunosurveillance mechanism by viral infection. PMID- 9395727 TI - A case of episodic angioedema associated with eosinophilia. AB - BACKGROUND: Gleich et al. first described 4 cases of episodic angioedema associated with eosinophilia as a distinct entity in 1984. Since then, several cases of this disorder have been reported in the United States, Europe and Japan. OBSERVATIONS: We report a case of a 22-year-old pregnant Japanese woman with this disorder. She had no fever and her general condition was good except the angioedema which was limited to her limbs. During an acute episode, her white blood cell count increased to 29,500/mm3 with 50% eosinophils, following an elevated serum interleukin-5 (IL-5) level. Spontaneous resolution occurred in 1 month after the onset. In a 5 month follow-up, no evidence of cardiac or other visceral organ involvement was found, and no recurrence occurred. CONCLUSIONS: Our case, combined with those reported in the literature, suggests that Japanese cases of episodic angioedema associated with eosinophilia differ from Caucasian cases in clinical symptoms and some other points. PMID- 9395728 TI - From molecular genetics to diagnosis and gene therapy. PMID- 9395729 TI - Genes involved in oncogenesis. PMID- 9395730 TI - Diagnostic molecular genetics. PMID- 9395731 TI - Gene therapy for the hemophilias. AB - There are many lines of evidence that suggest the eventual success of gene therapy as a treatment strategy for hemophilia. Because current treatment protocols using plasma-derived or recombinant proteins are far from ideal, the safe and efficient substitution of the defective gene by a normal copy of the gene, or at least its addition, would be of great benefit to the patient and may even be a potential cure. However, the construction of efficient gene therapy vehicles has proven quite difficult in the past and, so far, there is no system that promises to have all the desired features without any serious disadvantages. In general, either the levels of transgene expression are too low (because of the low titers achieved during the generation of the virus) or shortlived (e.g., because of the specific shut-off of the transferred promoter) as is often seen with retroviruses, or in the case of adenoviral vectors, expression is limited because of a strong immune response of the host. Clearly, much work remains to be done to optimize these promising though still imperfect vector systems. In the case of adenovirus, the development of less immunogenic vectors or in vivo modulation of the host immune system may hold promise for improvements. Reports by Yang et al. (1995) and Kay et al. (1995) are promising steps in the direction of immunomodulation. Both attenuate the immune reaction to the adenoviral vector by simultaneous application of either an interleukin or an immunoglobulin, respectively. When IL-2 was administered, the amounts of IgA were reduced and successful administration of a second dose of virus was possible. When CTLA4-Ig, an immunoglobulin that blocks the second signal during antigen presentation, was administered, a markedly prolonged expression of the transgene resulted. In vivo trials with AAV vectors have been carried out for some diseases (Flotte et al., 1993; Kaplitt et al., 1994) but not for hemophilia. Advances in high-titer AAV vector preparation will make this approach more feasible. The pace continues to quicken in the development of nonviral modes of gene delivery (Perales et al., 1994). Although these results are encouraging for the future of gene therapy as a treatment for genetic diseases, much work remains to be done to make this potential alternative a reality for treatment of hemophilia. PMID- 9395732 TI - Genes controlling retroviral virulence. PMID- 9395733 TI - Mapping animal genomes. PMID- 9395734 TI - The canine genome. PMID- 9395735 TI - The contribution of biological maturation to the strength and motor fitness of children. AB - The interrelationships among skeletal maturity, body size, strength and motor fitness were examined in American children 7-12 years of chronological age (CA). A total of 391 Black (184 boys, 207 girls) and 349 White (193 boys, 156 girls) children participated in the study. Biological maturity was assessed by the Tanner-Whitehouse II method, 20 bone skeletal ages (SA). Strength items included right and left grip strength, and pushing and pulling strength of the shoulders. Motor fitness items included a 35-yard dash, the standing long jump, and softball throw for distance. The standardized residuals of SA on CA (AG) were used to represent the effects of SA, independent of CA. Interaction terms were also computed by multiplying standardized values of stature (ST), body mass (MA), and AG together in all combinations. Regression analyses showed that the strongest predictor of strength was MA, while AG was the best predictor of motor performance. The interaction terms were also significant predictors of performance, explaining between 2% and 9% of the variance in 19 of the 41 significant regressions. The results highlight the complexity of the interrelationships among body size, biological maturation, strength and motor fitness. The effects of SA in children 7-12 years of age are expressed mainly through body size, but SA apparently influences motor fitness more so than muscular strength. PMID- 9395736 TI - Is there an independent association between parity and maternal weight gain? AB - The independent associations between parity and maternal body mass index (BMI), and between parity and maternal weight gain, were investigated using a combination of cross-sectional and longitudinal analyses based on a retrospective, repeat-pregnancy study that examined the change in maternal body weight from the beginning of one pregnancy to the beginning of the next. A group of 523 multiparous women who had been weighed regularly during pregnancy, and none of whom had fallen pregnant less than 12 months after the birth of their previous child, were examined. Sociodemographic, behavioural, medical, obstetric and perinatal data, together with antenatal measurements of maternal body weight and height, were abstracted from each mother's obstetric notes. Parity was found to be independently associated with maternal BMI (p < 0.001), gestational weight gain (p < 0.001) and interpregnancy weight gain (p = 0.032). Women of different parities were found to be at differential risk of long-term weight gain for two reasons. First, primiparous women are at risk of long-term weight gain because they gain the most weight during pregnancy, and high gestational weight gain is in itself a risk factor for long-term weight gain. Second, women of higher parity (4+) are at risk of long-term weight gain because they gain more weight in association with pregnancy, irrespective of the amount of weight they gain during their pregnancies. For women of parity 3 or less, the association between maternal body weight and parity appears to be the result of cumulative weight gained during successive pregnancies. For women of greater parity, the association between maternal body weight and parity is partly the result of cumulative excess gestational weight gained during successive pregnancies, and partly the result of gaining more weight from the beginning of one pregnancy to the next at later pregnancies. PMID- 9395737 TI - Nutritional status and age at menarche of Senegalese adolescents. AB - Growth and maturation during adolescence has not been well described in rural African populations, although it may represent the missing link between high levels of preschool stunting and nearly 'normal' adult heights. In 1995 the homes of subjects aged 10.3-17.5 years, living in a rural area of Senegal, were visited, and all adolescents present, 1527 boys and 1126 girls, were included in the analysis. A number of girls were absent because they worked in the capital city Dakar. Resident girls (n = 705) had significantly higher means than boys for all anthropometric variables (weight, body mass index, arm circumference and muscle arm circumference, triceps and subscapular skinfolds), except for height and head circumference. Girls who had just returned from seasonal migration to Dakar (n = 415) were, on average, 2 kg heavier, but not taller, than resident girls (p < 0.0001). The girls fell off in height from 11 to 13 years compared to the NCHS reference and then 'caught up' until the age of 17, while boys fell off during the entire age span. Mean age at menarche was estimated at 16.1 years (95% fiducial CI: 15.8-16.4) from status quo data by probit analysis. No significant difference was found between residents and migrants. Postmenarcheal girls had better nutritional status than premenarcheal girls in terms of height, weight, body mass index, percentage body fat and arm muscle circumference (p < 0.0001). In conclusion, puberty, as assessed by age at menarche, is delayed by about 3 years in this population, probably due to malnutrition. PMID- 9395738 TI - Body water distribution in highlanders versus lowlanders. AB - Acute exposure to high altitude produces characteristic changes in body water distribution from which acclimatized individuals seem to be spared. However, it has been suggested that body water distribution may be different in highlanders (HL) as compared to lowlanders (LL). We studied the distribution of total body water (TBW) between extracellular water (ECW) and intracellular water (ICW) in a group of 20 HL (3200 m above sea level) versus one of 20 LL (900 m above sea level). Subjects were matched for ethnic group (Kirghiz), sex (male), weight (Wt), height and body mass index. TBW:Wt and ECW:TBW were not different in HL as compared to LL (mean +/- SD, 58.5 +/- 5.0% versus 56.0 +/- 4.2% and 40.5 +/- 4.2% versus 40.7 +/- 2.2%; p = n.s. for both). This study does not support the hypothesis that body water distribution is different in HL as compared to LL. PMID- 9395739 TI - Methodological aspects of short-term knemometry in the assessment of exogenous glucocorticosteroid-induced growth suppression in children. AB - During recent years knemometry has been introduced for short-term assessment of the growth-suppressive effect of exogenous glucocorticosteroids in children. The aim of the present paper is to review methodological aspects of short-term knemometry used for this purpose. Knemometry has proven a highly accurate and reproducible method for assessment of short-term growth suppression in populations of children treated with exogenous glucocorticosteroids. Randomized, double-blind crossover and parallel designs applying consistent measurement intervals can be used. Confounding influences on the growth results from possible inter-group differences in spontaneous growth velocities are reduced in the crossover design. Glucocorticosteroid-induced knemometric growth suppression seems to reflect suppressive effects on soft tissue and bone components in the lower leg. In children treated with systemic glucocorticosteroids a shortening of the lower leg, which may be due to a reduction of the water content in the soft tissue, may confound the growth assessment. Suppressed short-term growth rates should be considered to have a poor correlation with long-term growth, though prospectively planned, controlled studies of the relation are needed in glucocorticosteroid-treated children. PMID- 9395740 TI - Anaemia- and malaria-attributable low birthweight in two populations in Papua New Guinea. AB - We studied 7300 singleton births in the highlands and 4881 in coastal Papua New Guinea in order to examine the separate contribution of anaemia or malaria to low birthweight. The highland sample was selected from a non-malarious area (Goroka) and the coastal sample from an area with perennial malaria transmission (Madang). There was an approximately three-fold increased risk of low birthweight (< 2500 g) in live-births in Madang compared to Goroka. The prevalence of anaemia in the two areas was strikingly different, with 29.2% of Goroka and 89.0% of Madang women anaemic. There was a trend towards increased low birthweight with decreasing haemoglobin levels in both areas, but this was significant only for Madang. It was assumed that for a given haemoglobin level the increased low birth weight percentage in Madang compared to Goroka was due to malaria exposure, and on this basis relative risk values were estimated for the effect of malaria exposure on low birthweight. Using this approach separate estimates for anaemia and malaria population-attributable risk for low birth weight in Madang were calculated. These indicated that up to 40% of low birthweight babies born in malarious areas may be attributable to malaria and less than 10% attributable to severe anaemia (Hb < 7.0 g dl-1). The magnitude of the malaria effect estimated in this analysis places a high priority on malaria control in pregnancy as a strategy for improving birthweight and child survival. PMID- 9395741 TI - Weight, height and arm circumference of children under 5 in the district of Mbarara, south-west Uganda. AB - This paper describes the growth of weight, height and arm circumference (MUAC) in children aged under 5 years and living in the south-west area of Uganda. The survey was carried out in 1988 and was based on a random sample of 31 villages of the Mbarara district. A total of 4320 children were measured by a team of 20 trained assessors. From these children a reference group was made up of the 3654 known to be still alive after 1 year. Growth charts were drawn by smoothing the non-parametric percentiles of the distribution of height, weight and MUAC for age and of weight for height. The anthropometric characteristics of children living in south-west Uganda differ considerably from those of children on which the FELS/NCHS/WHO references are based. Between 1 and 5 years of age, the median difference between Mbarara and American children increases from 1.5 to 3 kg for weight, from 4 to 7 cm for height, and from 1.5 to 2.5 cm for MUAC. These results imply that the use of the international reference may lead to low specificity and predictive values in screening malnourished children living in an underdeveloped country such as Uganda. The charts here proposed may apply to populations with a lifestyle similar to that of inhabitants of south-west Uganda, both from a nutritional and socioeconomic viewpoint. PMID- 9395742 TI - Longitudinal analysis of growth in children with idiopathic short stature. AB - In this study the growth curves of 229 children (145 boys, 84 girls ) with idiopathic short stature (ISS) were analysed in three ways: (1) we compared the results of longitudinal modelling by means of Karlberg's ICP model with those of a cross-sectional analysis; (2) we studied to what extent an individual changes his/her height standard deviation score (SDS) position during childhood; and (3) we constructed height velocity curves for children with ISS using Cole's LMS method, and compared these with the British references. During childhood the difference between the average longitudinal and the cross-sectional height curves was less than 1 cm and the spread was almost identical. The average change in height SDS during childhood was not different from zero, indicating that in general growth of children with ISS was well canalized. The individual change in height SDS position during childhood was within 0.6 SDS for a follow-up of 1 year, increasing to 1.9 SDS for a follow-up of 7 years. During childhood mean height velocity was about 1 cm/year lower than that of the British reference. With respect to the pubertal growth spurt the maximum height velocity took place 1 year later, and was 0.6 cm/year lower than the British reference in boys as well as girls. We conclude that spontaneous growth of children with ISS adequately described by a cross-sectional curve. PMID- 9395743 TI - Age variations in sibling correlations for height, sitting height and weight. AB - Familial correlations for height, sitting height and weight have been studied in a sample of 1278 siblings for the Biscay province (Basque Country), aged 4+ to 24+ years. The data have been internally standardized according to sex and age of individuals. The degree of resemblance among sibling has been expressed by intraclass correlation coefficients. The total sample has been divided into three age categories: < 12 years, 12-15 years, and > or = 15 years, in order to examine the effect of age on sibling correlations. In general, changes with age have been observed: sibling correlations for height show a clear upward trend through the considered growth period, reaching a value of 0.48 from 15 years of age. Intra correlations for weight show a slight downward trend with age. Sitting height shows a rather low correlation before 12 years of age, but equally high values in the other two ranges of age (0.48 and 0.47, respectively). This study confirms that the sibling resemblance for the analysed trait fluctuated through the growth period--height and sitting height showing similar patterns of variation with age- and that, after puberty, the degree of genetic determination is higher for bone measurements than for weight. PMID- 9395744 TI - Pathophysiology of portal hypertension. AB - Portal hypertension is a common clinical syndrome associated with chronic liver diseases and is characterized by a pathological increase in portal pressure. Increase in portal pressure is because of an increase in vascular resistance and an elevated portal blood flow. The site of increased intrahepatic resistance is variable and is dependent on the disease process. The site of obstruction may be: pre-hepatic, hepatic, and/or post-hepatic. In addition, part of the increased intrahepatic resistance is because of increased vascular tone. Another important factor contributing to increased portal pressure is elevated blood flow. Peripheral vasodilatation initiates the classical profile of decreased systemic resistance, expanded plasma volume, elevated splanchnic blood flow and elevated cardiac index. The elevated portal pressure leads to formation of portosystemic collaterals and oesophageal varices. Pharmacotherapy for portal hypertension is aimed at reducing both intrahepatic vascular tone and elevated splanchnic blood flow. PMID- 9395745 TI - Evaluation of patients with portal hypertension. AB - Patients with suspected portal hypertension must first be evaluated by physical examination, upper digestive endoscopy and ultrasonography with Doppler. Moreover, the evaluation of patients with portal hypertension depends on the cause of portal hypertension, the presence of complications and the specific treatment considered. Haemodynamic assessment with measurement of the hepatic venous pressure gradient is useful in confirming the origin of portal hypertension. This technique is the 'gold-standard' for evaluating haemodynamic treatments. Splanchnic and systemic circulation must also be measured. Quantitative evaluation of the splanchnic territory by Doppler sonography and other non-invasive investigations, may be performed. Further clinical studies are, however, needed to determine their interest in portal hypertension. PMID- 9395746 TI - Natural history. Clinical-haemodynamic correlations. Prediction of the risk of bleeding. AB - Promoting the development of oesophageal varices and ascites, portal hypertension dominates the clinical course of cirrhosis. Varices appear in patients with portal pressure gradient above 10 mmHg and enlarge in 10-20% within 1-2 years of their detection. Bleeding occurs in patients with portal pressure gradient above 12 mmHg when the wall tension causes the rupture of varices, with an incidence of about 10% per year. Indicators of bleeding risk are portal pressure gradient, variceal pressure, large varices and liver dysfunction. Mortality per bleeding episode is 30-50%. Among survivors 60% will rebleed and 30% will die in the following year. The risk of rebleeding decreases in patients with spontaneous or treatment induced reduction of portal pressure gradient or variceal pressure. Ascites develops in almost all patients along the course of the disease. Median survival after its appearance is less than 2 years. Less than 5% of cirrhotic patients die without ascites or without a previous bleeding. Thus portal hypertension is a major determinant of survival in cirrhosis. PMID- 9395747 TI - Portal Hypertensive gastropathy. AB - The term portal hypertensive gastropathy (PHG) defines a wide spectrum of diffuse macroscopic lesions that appear in the gastric mucosa of patients with portal hypertension. Histologically, these lesions correspond to dilated vessels in the mucosa and submucosa in the absence of erosions or inflammation. Endoscopically, the lesions are classified as mild when mosaic pattern or superficial reddening are present, and severe when gastric mucosa appear with diffuse cherry red spots. Mild lesions are highly prevalent (65-90%), whereas severe lesions are present in only 10-25% of cirrhotic patients. The pathogenesis of PHG is not well known, but both venous congestion related with raised portal pressure and increased gastric blood flow seem to be crucial factors for its development. Variceal sclerosis may contribute to the development or aggravation of the lesions. Bleeding is the unique clinical manifestation of PHG, and occurs only in those patients with severe lesions. During a 5-year follow-up, the risk of overt bleeding or chronic bleeding, which induces anaemia, is 60 % and 90%, respectively, for patients with severe PHG. Propranolol is the only pharmacological treatment that has been proven useful in preventing bleeding from PHG. Porto-systemic shunts and liver transplantation are also effective. PMID- 9395748 TI - Pharmacological prevention of variceal bleeding. New developments. AB - The introduction of pharmacological therapy has been one of the major advances in the treatment of the complications of portal hypertension. Many drugs have been shown to reduce portal hypertension in patients with cirrhosis. However, the most widely used drugs and the only ones for which there is sufficient evidence, are the beta-blockers. These drugs have been, up to now, the only accepted prophylactic therapy for oesophageal variceal bleeding and are also an alternative treatment to sclerotherapy or surgery to prevent variceal rebleeding. A reduction in portal pressure gradient by beta-blockers below 12 mmHg or by more than 20% of baseline values is associated with almost a total protection from oesophageal bleeding. Such a marked response in portal pressure is only achieved in some patients receiving propranolol. New pharmacological approaches with a greater portal pressure reducing effect may improve the beneficial effect of drugs in preventing variceal bleeding. The more promising approach is the combined administration of beta-blockers and isosorbide-5-mononitrate, which has been shown to potentiate the reduction in portal pressure and to be highly effective in initial randomized clinical trials. PMID- 9395749 TI - Endoscopic treatments for portal hypertension. AB - Endoscopic treatments for bleeding gastro-oesophageal varices include injection sclerotherapy, variceal obturation with tissue adhesives and variceal rubber band ligation. Today, endoscopic treatments are not recommended for the primary prophylaxis of variceal bleeding. Acute injection sclerotherapy remains a quick and simple technique for the control of active bleeding from oesophageal varices. Its efficacy may be improved by the early administration of vasoactive drugs. Banding ligation is the optimal endoscopic treatment for the prevention of rebleeding from oesophageal varices. The use of tissue adhesives and thrombin as injectates to treat bleeding fundal gastric varices and oesophageal varices not responding to vasoactive drugs or sclerotherapy is promising but needs further assessment by means of randomized controlled trials. PMID- 9395750 TI - Advances in drug therapy for acute variceal haemorrhage. AB - Recent advances in the pharmacology of portal hypertension are reviewed, against the background of existing knowledge and current clinical research. The most recent trials are analysed, and conclusions made about the use of drugs in acute variceal haemorrhage, as well as directions for further clinical trials and research. PMID- 9395751 TI - Transjugular intrahepatic portosystemic shunts (TIPS). AB - Transjugular intrahepatic portosystemic shunt (TIPS) is a procedure recently introduced for the management of complications of portal hypertension. TIPS can be placed in the liver with relative ease by a skilled radiologist with a low risk of mortality. The major complications following the procedure are infection, especially in patients undergoing emergency TIPS, intra-abdominal haemorrhage from capsular punctures, and long-term problems related to encephalopathy and stenosis of the shunt. Encephalopathy is more of a problem in older patients with wide diameter shunts. Stenosis of the shunt is related to pseudo-intimal hyperplasia, probably related to transection of bile ductules during placement of the shunt. In view of the high rate of encephalopathy and stenosis following the shunt, a careful follow-up of all patients, including ultrasonographic and angiographic examination of the shunt, is mandatory. TIPS is used predominantly for the control of acute variceal haemorrhage, prevention of recurrent variceal bleeding, and refractory ascites when conventional treatment has failed. However, the role of TIPS in the management of complications of portal hypertension still awaits the outcome of clinical trials. PMID- 9395752 TI - Surgery in portal hypertension. AB - The role of surgery in portal hypertension remains a topic of debate. For the past 100 years, various surgical procedures have been used to treat variceal bleeding, refractory ascites, and end-stage liver disease. The past decade has seen significant advances in pharmacotherapy, endoscopy, interventional radiology, and surgery for the management of patients with portal hypertension. Liver transplantation has come of age in the 1990s and is now an accepted therapy for patients with end-stage liver disease. The wide array of management options can complicate the decision making process and defines the need to evaluate these patients fully. Factors such as the aetiology and extent of liver disease, response to prior medical, endoscopic, and other interventional treatments, and possibility of future liver transplantation must be considered. This manuscript will review the history of surgical treatments of portal hypertension, describe the surgical procedures with their advantages and disadvantages, and evaluate their role in the elective and emergent settings. PMID- 9395753 TI - Ascites and renal functional abnormalities in cirrhosis. Pathogenesis and treatment. AB - In the past few years, there have been important advances in the field of pathogenesis and management of ascites and hepatorenal syndrome in cirrhosis. A new pathogenic theory of ascites and renal dysfunction in cirrhosis has been presented and previously ill-defined conditions, such as refractory ascites and hepatorenal syndrome, have been defined precisely. The link between the diseased liver and the disturbances in renal function and vasoactive systems is not completely known, but a large body of evidence indicates that it consists of a circulatory dysfunction that affects mainly the arterial circulation and is characterized by an inability to maintain an effective arterial blood volume within normal limits. The research on the mechanisms of this circulatory dysfunction will give valuable information in the design of more pathophysiologically oriented therapeutic approaches to the management of ascites. PMID- 9395754 TI - Hepatopulmonary syndrome: the paradigm of liver-induced hypoxaemia. AB - The current chapter deals with the concept, clinical manifestations and diagnostic tools of the hepatopulmonary syndrome (HPS) and highlights its most salient pathophysiological, mechanistic and therapeutic aspects. Defined as a clinical triad, including a chronic liver disorder, pulmonary gas exchange abnormalities and generalized pulmonary vascular dilatations, in the absence of intrinsic cardiopulmonary disease, this entity is currently growing in interest with both clinicians and surgeons. The combination of arterial hypoxaemia, high cardiac output with normal or low pulmonary artery pressure, and finger clubbing in a patient with advanced liver disease should strongly suggest the diagnosis of HPS. Its potential high prevalence together with failure of numerous therapeutic approaches depicts a life-threatening unique clinical condition that may dramatically benefit with an elective indication of liver transplantation (LT). A better orchestration of the concepts of the pathophysiology of this lung-liver interplay may foster our knowledge and improve the clinical management and indications of LT. PMID- 9395755 TI - Interferon therapy. PMID- 9395756 TI - Atypical esophageal ulceration. PMID- 9395757 TI - Kayexalate (sodium polystyrene sulphonate) in sorbitol associated with intestinal necrosis in uremic patients. AB - BACKGROUND: Kayexalate (sodium polystyrene sulphonate) in sorbitol is commonly used to treat hyperkalemia in patients with renal insufficiency. Isolated case reports and one recent large series have documented intestinal necrosis following administration of kayexalate in sorbitol. METHODS: Two patients with luminal kayexalate crystals associated with intestinal pathology were first identified in the pathology department, and clinicopathological correlation was carried out. RESULTS: Both patients were seriously ill, had prior cardiac surgery and were in renal failure (uremic). Examination of autopsy and colonic resection showed luminal kayexalate crystals associated with underlying mucosal necrosis, submucosal edema and transmural inflammation. CONCLUSION: Although occurring in complex clinical settings, the pathological findings provide additional evidence that kayexalate in sorbitol may be associated with intestinal necrosis and inflammation in uremic patients and that this may be a clinically and pathologically under-recognized iatrogenic bowel injury. PMID- 9395758 TI - A multicentre randomized controlled trial of recombinant interferon-alpha-2a in the treatment of patients with chronic hepatitis C. AB - Sixty-one chronic hepatitis C patients were randomly assigned to receive either 6 x 10(6) or 9 x 10(6) U of recombinant interferon-alpha-2a (IFN alpha-2a) six days a week for the first two weeks of treatment, followed in both cases by 6 x 10(6) U three days a week for the next 22 weeks. In the low dose group, 11 patients showed a complete response maintained for at least six months, 12 responded but then relapsed and nine did not respond; the corresponding figures in the high dose group were 10, 15 and five patients, respectively. The differences between groups are not statistically significant. Thus, this study provides no evidence of therapeutic benefit from increasing the initial dose of IFN alpha-2a. In both treatment groups, complete responders had significantly lower pretreatment viral titres than nonresponders and were significantly more likely to be infected by type 2a versus type 1b virus. PMID- 9395760 TI - Helicobacter pylori: from bench to bedside. AB - With the exponential increase in research in the field of Helicobacter pylori a paradigm shift has occurred. It is now recognized that H pylori is a chronic infection of the stomach causing inflammation. Some patients remain asymptomatic, while others may develop dyspepsia, duodenal or gastric ulcer, gastric cancer or a mucosa-associated lymphoid tissue lymphoma. However, the role of H pylori in contributing to nonulcer dyspepsia or nonsteroidal anti-inflammatory drug gastropathy remains controversial. An effective vaccine against H pylori is years away. Major interest has focused on the questions "who should be investigated and therefore treated" and "what is the latest gold standard for eradication of H pylori"? In Europe, guidelines have been developed to help the practitioner answer these important questions. Canadian guidelines will soon be available. For persons with known peptic ulcer disease there should be unequivocal acceptance that the good clinical practice of eradicating H pylori will result in substantial savings in health care expenses. The original 'classical triple therapy' (bismuth, metronidazole and tetracycline [BMT]) has now been surpassed by the combination of a proton pump inhibitor (PPI) plus two antibiotics (metronidazole plus clarithromycin; amoxicillin plus clarithromycin; or amoxicillin plus metronidazole), each given twice a day for one week. In Canada, the regimen of omeprazole plus one antibiotic (amoxicillin or clarithromycin) was approved recently but gives an eradication rate that is lower than the current target of 90%. According to the European (Maastricht) recommendations, if a single treatment attempt with PPI plus two antibiotics fails, PPI plus BMT is recommended. PMID- 9395759 TI - Intravenous cyclosporine for severe attacks of ulcerative colitis: a survey of Canadian gastroenterologists. AB - A single randomized trial evaluated the use of intravenous cyclosporine treatment for severe attacks of ulcerative colitis. The perceived efficacy and safety of this intervention were measured through a survey of the membership of the Canadian Association of Gastroenterology (CAG). METHODS: All CAG members were mailed a survey with questions regarding their familiarity with the data supporting the use of cyclosporine, their perception of the efficacy and toxicity of the drug, and whether patients who fail conventional treatment should receive this therapy. The proportion of respondents who had used cyclosporine to treat severe ulcerative colitis was determined. RESULTS: One hundred and sixty-one responses were received (34% response rate). Sixty-four per cent of respondents were academic faculty members and 82% treated patients with severe colitis. Using multivariate analyses, positive associations were found between the respondents' age (P = 0.004) and subspecialty training in gastroenterology (P = 0.001), and whether respondents treat patients with severe ulcerative colitis. Twenty-six per cent of individuals had prescribed cyclosporine for this indication, of whom 88% were in academic practice (P = 0.007). Over 90% of respondents believe that further clinical trials are needed before cyclosporine becomes accepted as standard therapy. CONCLUSIONS: Although the use of cyclosporine is measurable among Canadian gastroenterologists, the majority believe that further clinical trials are necessary before the drug is accepted as a standard therapy. PMID- 9395761 TI - Osteomyelitis and osteonecrosis in inflammatory bowel disease. AB - Osteomyelitis and osteonecrosis are skeletal disorders seen in patients with inflammatory bowel disease (IBD). Osteomyelitis usually occurs in the pelvic bones, especially in complicated Crohn's disease, presumably by direct extension from a pelvic inflammatory mass, abscess or fistulous tract. Diagnosis of osteomyelitis may be difficult and can lead to spinal extension of the septic process with a resultant neurological deficit, including paraplegia. Osteonecrosis or avascular necrosis has been reported in patients with either ulcerative colitis or Crohn's disease, often, but not exclusively, during or following steroid treatment. The disease is often multifocal, but its natural history is unknown, especially if diagnosed early with modern imaging methods, such as magnetic resonance. In IBD patients, the relationship between osteonecrosis and steroid use is unknown. An adverse steroid effect on bones, especially the femoral heads, may develop in some patients with IBD but, to date, this hypothesis remains unproven. Critical evaluation of published data reveals no consistent association between osteonecrosis and steroid treatment in IBD patients. PMID- 9395762 TI - Small bowel review: Part II. AB - Significant advances have been made in the study of the small bowel. Part II of this two-part review of the small bowel examines the early development and later ageing of the small bowel; the effect of diabetes, alcohol, radiation and HIV on the small bowel; enteral and parenteral nutrition; the brush border membrane and enterocyte proliferation; and peptide hormones (including transforming growth factors, motilin peptide YY and cholecystokinin). PMID- 9395763 TI - Hans Selye, inflammatory bowel disease and the placebo response. PMID- 9395764 TI - Colonic flora and fermentation: a new frontier for exploration. PMID- 9395765 TI - Liposome uptake by human retinal pigment epithelial cells in culture. AB - PURPOSE: To examine the internalization of liposomes containing various phospholipids by human retinal pigment epithelial (RPE) cells in culture. The internalization was compared to that exhibited by macrophages and fibroblast cells. METHODS: The uptake into cells was monitored by using a non-degradable, radioactive cholesterol-hexadecyl ether derivative and a pH-sensitive, water soluble fluorescent dye. Variables tested to determine their effect on uptake included time, phospholipid concentration, the presence or absence of serum in medium and the presence or absence of cholesterol in the liposome bilayer. The most avidly ingested liposome was tested to determine its ability to deliver a liposome-dependent drug. RESULTS: Liposome uptake was time and concentration dependent. Uptake for RPE was maximal in serum-free media, while the opposite was true for the other cells tested. Cell survival following exposure to fluoroorotic acid-containing liposomes correlated with the radioactive cholesterol-hexadecyl ether and fluorescent dye uptake data. CONCLUSIONS: No single liposome carrier or set of conditions tested appeared to be optimum for the delivery of a cytotoxic agent to these three cell types. This bears consideration when designing strategies for the prevention and treatment of intraocular proliferative diseases such as proliferative vitreoretinopathy (PVR). PMID- 9395766 TI - Quantitative study on regenerated retinal pigment epithelium and the effects of growth factor. AB - PURPOSE: To evaluate the integrity of damaged retinal pigment epithelium (RPE) with normal neural retina in a rabbit model, and to examine the effects of basic fibroblast growth factor (bFGF) on the damaged RPE. METHODS: A 3-port vitrectomy, a retinotomy, and a retinal detachment were made in pigmented rabbit eyes. The RPE was then abraded beneath the neural retina. In one group, 1 microgram(s) of bFGF was injected into the vitreous cavity (bFGF applied group). Zero, 7, 14, 21, and 28 days after the operation, fundus examination and fluorescein angiography were performed. Thereafter, the rabbits were killed, the eyes were enucleated, and the histological features were observed with light microscopy (LM), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The number of RPE cells in the treated area were counted in all experiments, and the effect of bFGF on the number of regenerated RPE cells was analyzed. RESULTS: Regeneration of the RPE was observed in the treated area at postoperative day 14. The number of regenerated RPE cells showed a linear increase. Regenerated cells observed by TEM were smaller than those of the normal RPE. The difference in the number of regenerated RPE cells between the bFGF-applied and non-applied groups was statistically significant on postoperative days 7 and 21. CONCLUSIONS: Repair of the RPE occurred in a short period of time. Similar RPE repair may be seen in humans. Intravitreal injection of bFGF increased the number of regenerated RPE in this model. PMID- 9395767 TI - Distribution of plasmalemmal Ca(2+)-pump and caveolin in the corneal epithelium during the wound healing process. AB - PURPOSE: Caveolae are small plasmalemmal invaginations which are assumed to play various physiological functions. In the present study, distribution of two caveolae-specific proteins, the plasmalemmal Ca(2+)-pump and caveolin, was examined in the corneal epithelium in the normal state and after artificial wounding. METHODS: A central epithelial ablation was made in the mouse cornea by a razor blade. After various intervals, the corneas were excised, fixed, and rapidly frozen. The specimens were subjected to immunofluorescence microscopy and immunoelectron microscopy, using antibodies against the plasmalemmal Ca(2+)-pump or caveolin. RESULTS: In the normal corneal epithelium, both plasmalemmal Ca(2+) pump and caveolin were observed along the cell surface by immunofluorescence microscopy, and were localized to caveolae by immunogold electron microscopy. In the regenerating epithelium, 12-18 h after injury, plasmalemmal Ca(2+)-pump was seen as many dots in the cytoplasm by immunofluorescence microscopy; in contrast, caveolin persisted along the cell surface. Immunoelectron microscopy revealed that the labeling for the plasmalemmal Ca(2+)-pump was located around membranous structures in the cytoplasm and was scarce along the plasma membrane, while caveolin remained in caveolae. The Ca(2+)-pump regained normal distribution when the wound was closed. By quantitation in electron micrographs, the number of caveolae per unit plasma membrane length was found to be decreased in the wounded corneal epithelium. CONCLUSIONS: The present results indicate that caveolae undergo compositional modification during the wound healing process of the corneal epithelium. Considering putative caveolar functions, the phenomenon may be related to possible fluctuations of the intracellular Ca(2+)-concentration in the regenerating epithelium. PMID- 9395768 TI - Computerized measurement of the three-dimensional distribution of optic disc pallor. AB - PURPOSE: To describe a method which provides quantitative measurements of the surface area of pallor in each quadrant of the three-dimensional optic cup, using photogrammetric measurements from simultaneous stereophotographs and computerized image analysis. METHODS: Simultaneous stereophotographs of one normal subject and two subjects with primary open angle glaucoma were digitized and analyzed for depth measurements. The boundaries of the optic disc, optic cup and region of pallor were identified. Pallor/disc and pallor/cup ratios were subsequently calculated for the superior, temporal, inferior and nasal walls. RESULTS: A digitized photograph and a Laplacian-filtered image were obtained for each eye to be studied. After processing each stereo pair through a similarity sequential detection-based algorithm, depth measurements are represented as a grey scale image, a contour plot, and a wire mesh, with the boundaries of the optic disc, optic cup and pallor superimposed. Ratios are given of the surface area of pallor to the surface area of the disc and the surface area of pallor to the surface area of the cup, by quadrant. CONCLUSIONS: Determination of surface area of pallor to cup may be useful in detecting early visual field loss in glaucoma and neurological disease. PMID- 9395769 TI - Effect of topical beta-blockers on tissue blood flow in the human optic nerve head. AB - PURPOSE: To study the effect of topical 0.5% timolol and 2% carteolol on tissue blood flow in the human optic nerve head (ONH). METHODS: Using a laser speckle tissue blood flow analyzer, normalized blur (NB), a quantitative index of tissue blood velocity, was measured every 0.125 s in the temporal site of the ONH free of visible surface vessels and averaged over 3 cardiac pulses (NBONH). To serve as a baseline, NBONH and intraocular pressure (IOP) in both eyes, blood pressure (BP) and pulse rate (PR) were recorded in healthy volunteers before, 1.5, 3 and 4.5 hrs after a 30L instillation of the vehicle of timolol or carteolol. From the following day and twice daily for 3 weeks, 30L of either 0.5% timolol or 2% carteolol was instilled into one eye and the respective vehicle into the fellow eye in a masked manner. NBONH, IOP, BP and PR were again recorded on the 21st and last experiment day. IOP was also recorded on the 7th and 14th days. Carteolol concentration in the plasma was also recorded after instillation of carteolol on the 21st day. RESULTS: During the baseline experiments, all the parameters recorded showed no significant change. After topical timolol, IOP was significantly reduced bilaterally with more reduction in the timolol-treated eye. Bilateral NBONH, BP and PR showed little change on the 21st day. After topical carteolol, IOP was significantly reduced bilaterally with more reduction in the carteolol-treated eyes on the 21st day. NBONH in the carteolol- and vehicle treated eyes was significantly higher on the 21st day than recorded in the same eye in the baseline experiment (P = 0.013 and 0.047), while BP and PR showed little change. The maximum carteolol concentration in plasma at 3 hrs on the 21st day averaged 1294 pg/ml. CONCLUSIONS: Results indicated that 3-week twice daily topical timolol treatment had no deleterious effect on the ONH tissue blood flow in the human eye, and that 3-week twice daily topical carteolol treatment may increase the tissue blood flow in the human ONH. PMID- 9395770 TI - Effect of shaking of corneal endothelial preservation. AB - PURPOSE: To investigate the effect of shaking on corneal endothelial preservation. METHODS: Thirty-six dog corneal buttons were obtained under standard eye bank procedure, after the animals had been sacrificed for cardiovascular experiments in the surgical department. They were equally divided into two groups. In Group I, buttons were put in Dexsol preservative medium and preserved at 4 degrees C. In Group II, buttons were put in Dexsol preservative medium and shaken in a shaking incubator at a speed of 5 rpm. at 4 degrees C for 10 h. After shaking, they were returned to a 4 degrees C refrigerator until examination. Three buttons from each group underwent specular microscopy, scanning electron microscopy (SEM), and alizarin red with trypan blue stain examinations on days 0, 1, 3, 5, 7 and 9, respectively. RESULTS: In Group I, intercellular borders became blurred under specular microscopy, beginning on day 5. However, endothelial cell count did not change significantly until day 9. Intercellular digitations and wrinkling of intercellular borders were seen under alizarin red with trypan blue stain and SEM examination, beginning on day 3. Pleomorphism and ill-defined intercellular junctions were seen under SEM on day 7. There was no obvious denudement of the endothelial sheet, but a few scattered exfoliations were seen in Group I. In Group II, endothelial cell count did not decrease on day 1, but an endothelial image could not be obtained by specular microscopy 3 days after treatment. Alizarin red with trypan blue stain and SEM examination revealed that the endothelial cells were denuded from the Descemet's membrane three days after shaking. Severe stromal edema was seen under SEM five days after shaking. CONCLUSIONS: The results showed that shaking had a detrimental effect on endothelial cell preservation. Vigorous shaking should be avoided during transportation of corneal buttons. It is advisable to perform penetrating keratoplasty as soon as possible upon receiving transported donor corneas. PMID- 9395772 TI - Immunodetection of membrane skeletal protein 4.2 in bovine and chicken eye lenses and erythrocytes. AB - PURPOSE: Protein 4.2 is a major erythrocyte membrane skeletal protein, playing an important role in maintaining the integrity and stability of the membrane. It is a transglutaminase-like molecule with no enzymatic cross-linking activity. Several protein 4.2-associated proteins (i.e. band 3, ankyrin, and protein 4.1) and transglutaminase activities have been detected in the lens. The purpose of this study is to find out if protein 4.2 is also expressed in lens fiber membranes. METHODS: Western blot analysis of cell membranes isolated from bovine and chicken lens fibers and erythrocytes, and immunocytochemistry of frozen sections of bovine and chicken lens fibers were carried out using two protein 4.2 specific antibodies. These two peptide antibodies have been used to identify two alternatively spliced protein 4.2 isoforms in human erythrocyte membranes: the short (P4.2S, or hP4.2(691)) and the long (P4.2L, or hP4.2(721)) isoforms. RESULTS: Western blot analysis using anti-P4.2(L) antibody demonstrated specific immunoreactive polypeptides in bovine and chicken lens fiber membranes and erythrocyte membranes, co-migrating with hP4.2(721). Immunofluorescence staining of bovine and chicken lenses, using anti-P4.2(L) antibody, revealed specific signals along the cell membranes of cortical fibers. The signals exhibited a unique, patchy pattern along the cortical fiber cell membranes in both cross sectional and longitudinal views. In cross sections, the labeling of anti-P4.2(L) along the entire cell membranes gave an appearance of a hexagonal shape of fiber cells. CONCLUSIONS: Protein 4.2, or its analogs, is present in the lens fiber membranes. Its specific staining pattern in the lens fibers suggests that it participates in the architecture of the lens fiber cell membranes, and may play a role in the lens mechanics and pathology. PMID- 9395771 TI - Inhibition of RPE cell-mediated matrix adhesion and collagen gel contraction by crovidisin, a collagen-binding snake venom protein. AB - PURPOSE: Cell-mediated collagen gel contraction plays an important role in the pathogenesis of proliferative vitreoretinopathy (PVR). Anti-adhesion therapy has been suggested as a promising strategy in the treatment of PVR. Crovidisin, a snake venom protein isolated from Crotalus viridis, has been shown to bind selectively to collagen and to inhibit collagen-induced platelet aggregation. In the present study, the effectiveness of crovidisin in inhibiting the attachment of retinal pigment epithelial (RPE) cells to collagen, and RPE cell-mediated collagen gel contraction, was evaluated. METHODS: Fluorescein isothiocyanate (FITC)-conjugated crovidisin was prepared and used to evaluate its binding affinity for collagen type I, fibronectin, vitronectin, and laminin. The inhibitory effect of crovidisin on RPE cell-mediated extracellular matrix attachment and collagen gel contraction was evaluated by cell adhesion and type I collagen gel contraction assays. The cytotoxic effect of crovidisin was examined with a cell proliferation assay, using the Alamar blue method. Flavoridin, an Arg Gly-Asp-containing peptide from viper venom, was used for comparison. RESULTS: FITC-conjugated crovidisin bound selectively to collagen type I with high affinity. It did not bind to other matrix proteins, including fibronectin, vitronectin and laminin, nor to RPE cells. Crovidisin inhibited RPE cell attachment to type I collagen in a dose-dependent manner. This inhibitory effect was enhanced by the presence of flavoridin. Crovidisin also dose-dependently inhibited RPE cell-mediated type I collagen gel contraction. Crovidisin was non toxic to RPE cells. CONCLUSIONS: Crovidisin, a snake venom-derived collagen binding protein, possessing an inhibitory activity on RPE cell-collagen interaction and RPE cell-mediated collagen gel contraction, may be a useful tool for studying cell-collagen interaction, and a potential anti-adhesion therapeutic agent for ocular disorders in which cell-collagen interaction in involved, such as PVR. PMID- 9395773 TI - A correlation between computer-predicted changes in secondary structure and the phenotype of retinal degeneration associated with mutations in peripherin/RDS. AB - PURPOSE: To investigate a molecular understanding of how mutations can lead to different phenotypes, we analyzed the relationship between altered secondary structures predicted by missense mutations in the peripherin/RDS and clinical severity of autosomal-dominant retinal degeneration. METHODS: We analyzed thirteen different kinds of missense mutations in the second intradiscal loop of peripherin/RDS, previously reported in peer review journals. Alteration of the secondary structure of peripherin/RDS was predicted by computer-assisted protein structure analysis. The number of amino acid residues that would be involved in the secondary structural change produced by a given missense mutation was scored as a grade of molecular change. Clinical severity was estimated by the impairment based on electroretinographic recordings of rods and cones, and was scored according to the severity of their recordings. Regression analysis was carried out between both scores of molecular change and clinical severity. Effects of patients' ages on clinical severity was also analyzed. RESULTS: Significant correlation was found between scores of molecular change and those of clinical severity (rods, r = 0.89, p < 0.001; cones, r = 0.76, p < 0.005) by regression analysis. There was no correlation between clinical severity and patients' ages. CONCLUSION: . These findings indicate that the degree of change in the secondary structure of peripherin/RDS can explain in part the correlation between genotype and phenotype in autosomal-dominant retinal degeneration associated with missense mutations in the peripherin/RDS gene. PMID- 9395774 TI - Radiation response of porcine RPE cells in vitro. AB - PURPOSE: To investigate the antiproliferative effect of ionizing radiation on retinal pigment epithelial (RPE) cells that are supposed to play a major role in the pathogenesis of proliferative vitreoretinopathy (PVR). METHODS: RPE cells from pig eyes were irradiated with doses ranging from 4 to 16 Gy (1 Gray = 1 Joule/kilogram). Cells were counted at 1, 2, 3, and 4 weeks (Experiment 1) or 1, 2, 4, and 6 weeks (Experiment 2) after treatment. In Experiment 3, cells were trypsinized 24 h after radiation and seeded again. Colonies were counted 10 days later, and the surviving fraction was determined. RESULTS: The numbers of cells and colonies were inversely correlated to the doses applied. In Experiment 2, cell numbers of radiated cultures remained stable during the time of follow-up, whereas, in Experiment 1, significant proliferation occurred in treated cultures as well as in controls. This may be due to the higher growing rate that was found in the cultures of Experiment 2, compared to those of Experiment 1, at the time of radiation. In Experiment 3, a D0 value of 0.72 Gy was found. CONCLUSIONS: Proliferation of RPE cells can be suppressed by irradiation in a dose-dependent manner. Therefore, radiotherapy may be useful in the treatment of PVR. Its effect probably depends on the stage or activity of PVR at the time of radiation. PMID- 9395776 TI - MHC-II but not MHC-I responses are required for vaccine-induced protection against ocular challenge with HSV-1. AB - PURPOSE: To determine the importance of major histocompatibility complex (MHC) class-I versus MHC class-II immune responses in protecting naive versus vaccinated mice against an ocular HSV-1 challenge. METHODS: Class-II deficient A beta o/o (CD4-CD8+ T cells) knockout mice, which are effectively CD4+ T cells negative, and class-I deficient beta(2)mo/o (CD4+CD8- T cells) knockout mice, which are effectively CD8+ T cells negative, were either vaccinated or mock vaccinated and examined for their ability to withstand HSV-1 ocular challenge. RESULTS: Unvaccinated A beta o/o and beta(2)mo/o mice were both more susceptible to lethal ocular HSV-1 infection than the parental wild type C57BL/6J mice, indicating that both MHC-I and MHC-II were required for optimal protection of naive mice against ocular HSV-1 challenge. Vaccinated beta(2)mo/o mice produced significant neutralizing antibody titers, and following ocular challenge, these mice were completely protected against death and corneal scarring. In contrast, vaccinated A beta o/o mice developed no neutralizing antibody titers and vaccination did not provide these mice with any protection against death or corneal scarring. Passive transfer of anti-HSV-1 antibody into A beta o/o mice up to 6 days post ocular challenge resulted in complete protection against death and corneal scarring. CONCLUSIONS: Passive antibody transfer, but not vaccination, protected A beta o/o mice against ocular challenge. In contrast, vaccination completely protected beta(2)mo/o mice. These results suggest for a vaccine to provide optimal protection against ocular HSV-1 challenge in this system, it is not only sufficient, but it is also required, that the vaccine induce an effective neutralizing antibody response. PMID- 9395777 TI - Fluorescein as a marker for subretinal transplantation of human fetal neural retina. AB - PURPOSE: To investigate the effect of fluorescein on human fetal neural retina and adult rat retina; and to use fluorescein to map the area of subretinal transplantation. METHODS: In vitro: Human fetal neural retina (8 to 14 weeks gestational age) was incubated in 0.03% fluorescein in Dulbecco's Modified Eagles Medium (DMEM) or DMEM alone for 30 min. Viability was determined using the trypan blue exclusion test, and results were compared. Effects of the fluorescein on cell morphology were assessed by observation of primary cultures for 1 week. In vivo: Human fetal neural retina was mechanically dissociated in 0.03% fluorescein in DMEM and transplanted to the subretinal space of immunosuppressed rats. To control for the effect of fluorescein on the grafted tissue, transplants were also performed in DMEM only. After transplantation, indirect ophthalmoscopy and true color fundus photography were performed to document the area covered by the transplant. One month after transplantation, the appearance of grafts exposed to fluorescein was compared to those that were not, at the light microscopic level. RESULTS: In vitro: Exposure of human fetal neural retina to fluorescein had no effect on viability. Similarly, in tissue culture, the fluorescein-exposed cells exhibited the same phenotype as the controls. In vivo: Immediately after transplantation the graft site was clearly outlined within the subretinal area and fluoresced intensely. There were no traces of the dye 2 h after transplantation. Cells that were transplanted with fluorescein survived transplantation, and one month after transplantation could be seen forming subretinal grafts. No differences were noted between these and control grafts. CONCLUSIONS: Fluorescein is an effective dye for immediate and transient localization of trans-scleral transplants to the subretinal space. It allows mapping of the area covered by the injection without interfering with the viability and differentiation of the transplanted cells. It allows unequivocal photo- and video-documentation in both the albino and pigmented fundi. It is already FDA approved for many other extra- and intraocular studies and now has directly been shown to be non-toxic to both human fetal neural retina and adult rodent retina. PMID- 9395775 TI - Induction of rabbit cyclooxygenase 2 in the anterior uvea following glaucoma filtration surgery. AB - PURPOSE: This study was undertaken to evaluate for the presence of cyclooxygenase 2 (COX2) gene expression in the anterior uvea of rabbits following glaucoma filtration surgery. METHODS: One of the following surgical procedures were performed on the right eye of New Zealand white albino rabbits: (1) paracentesis (2.5 mm limbal incision); (2) iridectomy through a 2.5 mm limbal incision; (3) lamellar scleral flap formation or (4) full glaucoma filtration surgery. The animals were sacrificed within 3 hours of post-surgery, and the anterior uveal tissues were isolated. Polymerase chain reaction-based techniques were employed to assay for the presence of COX2 transcript. RESULTS: A partial coding sequence of the previously unreported rabbit COX2 gene was obtained. COX2 mRNA was detected in the operated eyes of animals that underwent either full filtration surgery or iridectomy through a limbal incision. CONCLUSIONS: In normal rabbit anterior uveal tissue, there appears to be minimal expression of COX2 message. After experimental glaucoma filtration surgery, there is rapid induction of COX2 message. PMID- 9395778 TI - Physostigmine increases aqueous humor production in human eyes. AB - PURPOSE: To investigate if part of the progressive reduction of intraocular pressure (IOP), seen when physostigmine is applied on alternate hours, is due to a reduced aqueous flow. METHODS: In a randomized, open study, one drop of physostigmine salicylate, 8 mg/ml, was instilled at 7 AM in one randomly assigned eye in each of twenty healthy volunteers. Instillations were repeated on alternate hours throughout the day. Each subject's untreated eye served as control. Fluorophotometry of the anterior segment was performed hourly between 7 Am and 8 PM and aqueous flow was calculated. Subsequently, the subjects underwent tomography and tonometry. The change in anterior chamber depth and volume induced by physostigmine was assessed separately. RESULTS: The mean aqueous flow during the day was 25-28% higher in the physostigmine-treated eye than in the control eye. The difference was statistically significant from 9 AM (p < 0.05-p < 0.001). Each dose caused a further increase. The mean outflow facility increased by 0.14 microliters/min/mm Hg with 95% confidence interval (CI) 0.09-0.18. Although the increase in outflow facility was small, there was a marked reduction of IOP with a mean difference between treated and untreated eye of 3.2 mm Hg (95% CI: 2.3 4.0). CONCLUSIONS: Repeated administrations of physostigmine increase the aqueous flow and outflow facility. The combined effect is a marked reduction of IOP. PMID- 9395779 TI - Induction of donor-specific ACAID can prolong orthotopic corneal allograft survival in "high-risk" eyes. AB - PURPOSE: To examine the effect of donor-specific anterior chamber-associated immune deviation (ACAID) induction on the survival of orthotopic corneal allografts in neovascularized graft beds. METHODS: To induce donor-specific ACAID in recipients, peritoneal exudate cells (PEC) from C57BL/6 mice were incubated overnight with transforming growth factor (TGF)-beta. Cultured PEC were injected intravenously (i.v.) into BALB/c mice, and, 1 week later, these animals received orthotopic corneal allografts from C57BL/6 donors into neovascularized graft beds. Control mice received i.v. injection of syngeneic (BALB/c) PEC, cultured overnight with TGF-beta, and then received orthotopic corneal allografts from C57BL/6 donors. RESULTS: All corneal allografts (15 out of 15) were rejected within 2 weeks after grafting in the neovascularized graft beds of control animals. However, only 6 out of 16 (37.5%) of corneal allografts were rejected in recipients in which donor-specific ACAID had been induced by injection of allogeneic PEC cultured with TGF-beta. CONCLUSION: Previous studies revealed that rejection of orthotopic corneal allografts in neovascularized graft beds in mice correlated with acquisition of donor-specific delayed hypersensitivity (DH). The results of this study suggest that induction of donor-specific ACAID, which selectively impairs DH responses to donor antigens, effectively prolongs corneal allograft survival in "high-risk" eyes. PMID- 9395780 TI - Myocardial contrast echocardiography. AB - MCE has evolved from a laboratory tool to a clinical procedure. It would be wrong to consider it merely another tool for imaging of myocardial perfusion. As discussed, it allows physicians to bring physiology and pathophysiology to the bedside, providing a better understanding of the underlying mechanisms of abnormal findings in individual patients. MCE can provide quantitative measurements that can be repeated as often as necessary in a patient. Because of its complexity, large clinical studies are necessary to define the role of MCE in the general clinical milieu. Advances in MCE continue at a very rapid pace, and its potential for the study of endothelial function, site-specific targeting, and local delivery of drugs appears promising. Its role will continue to evolve into the early part of the next century. What we learn of the myocardium can be easily applied to other organ systems accessible to ultrasound. The future of MCE appears very exciting. PMID- 9395782 TI - The diaphragm in chronic obstructive pulmonary disease: how useful is it? PMID- 9395781 TI - Intestinal taurine transport: a review. AB - Intestinal uptake of dietary taurine is an important contributor to taurine homeostasis and may become crucial when taurine metabolism is impaired. This review aims to assess the literature documenting taurine transport and review what is currently known about the operation of the enterocyte taurine transport protein. Sources included MedLine searches from the last 10 years and references from original and review articles. The aim was to include human and animal studies directly addressing the subject of taurine uptake by enterocytes. Intestinal taurine transport has been well documented in in vivo studies using many different animal models. The mechanistic/kinetic aspects of the transport system have been extensively documented. However, little is known about what regulates the system. The recent development of a cell culture model of intestinal taurine transport will allow studies to explore the regulation of gut taurine uptake, which promises to be a very exciting area. PMID- 9395783 TI - Helicobacter pylori and non-Helicobacter pylori bacterial flora in gastric mucosal and tumour specimens of patients with primary gastric lymphoma. AB - There is an association between Helicobacter pylori (H. pylori) and gastric mucosa-associated lymphoid tissue (MALT) and MALT lymphoma. Histologically, mainly non-specific stains are used to detect H. pylori, such as haematoxylin eosin (HE) or modified Giemsa (MG). In this study, both a MG and a specific immunohistochemical stain (IMM) for H. pylori (Dako B471) were performed on sequential slides of resected material containing tumour and non-tumorous gastric mucosa from patients with primary gastric lymphoma (n = 52). Special attention was paid to the presence of non-H. pylori bacterial flora diagnosed by a positive MG (according to form and localization) and a subsequently negative IMM. On all slides, bacterial density was scored semiquantitatively (grades 0, 1, 2, 3). In total, 32 (61.5%) patients were H. pylori positive using IMM and 34 (65.4%) were non-H. pylori positive using MG. In 24 out of the 34 patients, the non-H. pylori flora consisted mainly of cocci in combination with rods in 15 patients, mostly in minor quantities; in another 10 patients, high numbers of both cocci and different types of rods were present. Most non-H. pylori bacteria were localized superficially, although in 22 patients minor quantities of non-H. pylori were also seen in the glandular lumina. After all of the patients had been analysed, no differences in the density of H. pylori and of non-H. pylori flora were found. Only when comparing patients who had a small-cell lymphoma with those who had a large-cell lymphoma was a significantly higher density of H. pylori found in the corpus mucosa of large-cell lymphomas and a higher prevalence of non-H. pylori was found in tumours, in antrum or corpus, of patients with large-cell lymphomas. In conclusion, with joint evaluation using MG and a H. pylori-specific immunohistochemical stains, the proportion of H. pylori-positive gastric lymphoma patients was lower than in most previous studies but other bacteria were found in a relatively high proportion. The role of the non-H. pylori intragastric bacterial flora identified in this study has to be further elucidated in the aetiopathogenesis of primary gastric lymphoma. PMID- 9395785 TI - Hyposensitivity to nerve stimulation in portal hypertensive rats: role of nitric oxide. AB - Portal hypertension goes along with vascular hyporeactivity, partly mediated by nitric oxide (NO). Interactions between the adrenergic nervous system and NO in portal hypertension are undetermined. We tested (1) whether superior mesenteric arterial beds of portal hypertensive rats have an altered sensitivity to periarterial nerve stimulation (PNS) and (2) the role of NO in modulating nerve stimulated responses. Vasopressor responses to PNS (Hz, 2-32) were similar in preparations of partial portal vein-ligated (PVL, n = 12) and control (CON, n = 12) rats (60.0 +/- 6.7 and 47.8 +/- 6.1 CmH2O respectively) for 24 Hz (NS), but sensitivity of vessels of portal hypertensive animals displayed a significant rightward shift [Hz needed for 50% of maximal response (HZ50) being 15.5 +/- 0.4 and 12.9 +/- 0.6 for PVL and CON respectively, P < 0.001]. NO formation inhibition by N omega-nitro-L-arginine (10(-4) mol L-1) significantly increased responses to PNS (P < 0.05), the absolute values for 24 Hz being 101.4 +/- 11.7 cmH2O for PVL (n = 8) and 86.4 +/- 11.4 cmH2O for CON (n = 7) (NS). NO formation inhibition reversed the hyposensitivity in preparations of PVL, Hz50 being 13.9 +/- 0.5 and 13.2 +/- 0.2 for PVL and CON respectively (NS). Adrenergic receptor antagonism with prazosin (10(-7) mol L-1) and yohimbine (10(-6) mol L-1) inhibited PNS-mediated vasopressor reactivity (n = 6 per group, P < 0.001), confirming the nervous origin of vasoconstrictor responses. It is concluded that (1) portal hypertension goes along with a significant hyposensitivity to PNS and (2) this hyposensitivity is reversed by NO-formation inhibition PMID- 9395784 TI - Signal transduction pathways of membrane expression of proteinase 3 (PR-3) in human endothelial cells. AB - At present, the exact mechanism of the pathogenic effect of anti-PR-3 antibodies remains unknown. Interaction of anti-neutrophil cytoplasmic antibodies (ANCAs) with human umbilical vein endothelial cells (HUVECs) may play a key role. Recently we were able to show that ANCAs recognize their target antigen, PR-3, translocated into the membrane of HUVECs. The objective of this study was to investigate regulation, i.e. signal transduction pathways, of PR-3 expression in endothelial cells. HUVECs were isolated according to the method of Jaffe et al. and cultured under standard conditions. A cyto-enzyme-linked immunosorbent assay (ELISA) with unfixed cells was performed. Membrane-expressed PR-3 was detected by affinity-purified and monoclonal anti-PR-3 Ab. Tumour necrosis factor alpha (TNF alpha)-induced membrane expression of PR-3 could be blocked with the RNA synthesis inhibitor actinomycin D, the protein kinase C (PKC) and proteinase A (PKA) inhibitor staurosporine, the specific PKA inhibitor calphostin C, the c-AMP dependent PKA inhibitor KT5720 and the tyrosine kinase inhibitor genistein in a dose-dependent manner. The effect of calphostin C was the most significant. In addition, the effect of phorbol 12-myristate 13-acetate (PMA), a mediator of intracellular second messengers, was investigated. In our study, pretreatment of cells with PMA for 48 h led to a down-regulation of PR-3 expression. This effect, however, could be overridden by TNF-alpha stimulation, i.e. TNF-alpha-induced membrane expression of PR-3 was resistant to down-regulation of PKC. In conclusion, our data suggest that translocation of PR-3 in HUVECs is an active process depending on protein synthesis. PR-3 expression by HUVECs may involve a PKC reactive to cytokines such as TNF-alpha which induces PR-3 expression at a transcriptional level. PMID- 9395786 TI - Phyllanthus amarus suppresses hepatitis B virus by interrupting interactions between HBV enhancer I and cellular transcription factors. AB - The Phyllanthus amarus plant suppresses HBV mRNA transcription in vitro and exhibits therapeutic potential in chronic HBV carriers, although further work is necessary to define its mechanism of action. Analysis in HuH-7 cells with transfected plasmids using a luciferase reporter showed that P. amarus specifically inhibited HBV enhancer I activity. To identify the mechanism of this HBV enhancer I inhibition, liver-enriched cellular transcription factors were co expressed in HuH-7 cells. The C/EBP alpha and beta, as well as HNF-3 alpha and beta transcription factors, significantly up-regulated the HBV enhancer I activity. In contrast, co-transfection of HNF-I alpha or beta had no effect upon the HBV enhancer I activity. Exposure to P. amarus inhibited C/EBP alpha- and beta-mediated up-regulation of HBV enhancer I activity in a dose-dependent manner, whereas HNF-3 alpha- and beta-mediated up-regulation of HBV enhancer I was unaffected. In vitro gel shifts showed that P. amarus inhibited complexing of C/EBP transcription factors to a consensus oligonucleotide sequence, whereas DNA binding of AP-1 and SP-1 transcription factors was unaffected. As P. amarus down regulates HBV mRNA transcription by a specific mechanism involving interactions between HBV enhancer I and C/EBP transcription factors, purification and further analysis of the active P. amarus component will advance insights into its antiviral activity. PMID- 9395787 TI - Circulating endothelial cell markers in peripheral vascular disease: relationship to the location and extent of atherosclerotic disease. AB - We examined the relationship between specific endothelial cell markers soluble E selectin, von Willebrand factor and soluble thrombomodulin and the location or extent of atherosclerosis by analysing plasma samples from 200 patients with symptomatic peripheral vascular disease and 213 age- and sex-matched asymptomatic control subjects. Using ELISAS, we found increased von Willebrand factor and thrombomodulin (both P < 0.0001) in the patients relative to the control subjects, but no significant change in soluble E-selectin. Soluble thrombomodulin was increased in patients with disease at one locus (i.e. of the carotid or iliac/femoral arteries), with an additional significant increase in patients with disease at multiple loci (i.e. any combination of carotid, coronary or iliac/femoral artery disease). No marker differentiated carotid artery disease from iliac/femoral artery disease. We conclude that von Willebrand factor is a marker of generalized atherosclerosis, but that soluble thrombomodulin is related to the extent of disease. Further research into these endothelial cell products are warranted to explore their diagnostic and/or prognostic potential. PMID- 9395788 TI - Detection and quantification of the control proteins of the alternative pathway of complement in 3T3-L1 adipocytes. AB - The complement peptide C3a desarg is identical to acylation-stimulating protein (ASP), a human plasma protein that potently stimulates adipocyte triacylglycerol synthesis and glucose transport. Both human and murine adipocytes express mRNA and/or protein for the complement components C3 and factors B and D (adipsin) required to generate ASP. However, the regulatory mechanisms controlling this process are unknown. We have established a semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR) technique to demonstrate the presence in mouse 3T3-L1 adipocytes of mRNA for all components of the alternative pathway, including the control proteins factors I and H, CR1 and properdin. On differentiation, mRNA for C3 (fivefold) and factor D (> 50-fold) increased, whereas stimulation with tumour necrosis factor (TNF)-alpha and interleukin (IL) 1 beta led to eightfold increases in factor B mRNA. Metabolic labelling followed by immunoprecipitation showed that factor B protein is normally present in small quantities, and is greatly increased by cytokine stimulation. The larger quantities of C3 and H proteins present were little affected, whereas levels of C3a increased on cytokine stimulation. These results suggest that the rate limiting step in the cytokine-induced production of ASP in adipocytes is factor B synthesis. PMID- 9395789 TI - The Asn-291-->Ser and Ser-477-->Stop mutations of the lipoprotein lipase gene and their significance for lipid metabolism in patients with hypertriglyceridaemia. AB - We examined 99 Finnish patients whose serum fasting triglycerides (TG) had exceeded 6.0 mmol L-1 with special interest to their lipid, lipoprotein and post heparin plasma lipase activities. The control group consisted of 75 healthy individuals. We also determined the frequency of the Asn-291-->Ser and Ser-447- >Stop mutations both in hypertriglyceridaemic (HTG) subjects and in control subjects. A total of 51 of the original 99 hypertriglyceridaemic patients still had TG > 6.0 mmol L-1 when measured a second time. They are referred to as persistently hypertriglyceridaemic subjects (pHTG). The remaining 48 subjects had TG < 6.0 mmol L-1 in the second measurement and are referred to as sporadically hypertriglyceridaemic subjects (sHTG). The allelic frequencies of the Ser-447- >Stop mutation in the total HTG and sHTG groups were similar to the frequencies present in the control group, but lower in pHTG patients compared with the control group (0.049 vs. 0.153, chi(2) = 6.63, P < 0.05). The Asn-291-->Ser mutation was more frequent in HTG group than in the control group (0.0606 vs. 0.013, chi(2) = 4.86, P < 0.05). This difference was due to the higher frequency of the minor allele of Asn-291-->Ser in the cohort with persistent hypertriglyceridaemia compared with the control group (0.088 vs. 0.013, chi(2) = 8.00, P < 0.01). The highest frequency (0.114) of the minor allele of Asn-291- >Ser was found in type 2 diabetic patients with persistent hypertriglyceridaemia. The carrier status of Asn-291-->Ser or Ser-447-->Stop did not predict either post heparin plasma lipoprotein lipase (LPL) activities or lipid and lipoprotein levels in any of the groups studied. Our data suggest that overproduction of very low-density lipoproteins (VLDL) is a more important cause of hypertriglyceridaemia in the Finns than is the LPL deficiency. PMID- 9395790 TI - The role of intravenous administration of dextran 70 in enteric bacterial translocation after partial hepatectomy in rats. AB - The aim of this study was to assess the effect of intravenous dextran on bacterial translocation and intestinal vascular endothelial and epithelial barrier function after experimental partial hepatectomy. We determined systemic arterial pressure, enteric bacterial growth (proximal and distal small intestine and colon) and bacterial translocation (BT) to mesenteric lymph nodes (MLN), liver, lungs, spleen, kidneys and blood, as well as intestinal vascular endothelial and epithelial barrier permeability, after sham operation or partial hepatectomy (50% and 90%) with preoperative intravenous administration of saline, albumin or dextran 70. Subtotal hepatectomy induced a significant decrease in arterial pressure and an increase in the number of Escherichia coli in the distal small intestine. BT was not observed in sham-operated animals or in rats with 50% hepatectomy administered dextran. The number of positive cultures of enteric bacteria was significantly increased after hepatectomy, whereas dextran treatment decreased the number of animals with BT. Increased permeability of the intestinal vascular endothelial and epithelial barriers was noted in hepatectomized animals, while dextran prevented hepatectomy-induced vascular endothelial barrier injury. Enteric bacterial translocation occurred following partial hepatectomy in the rat, associated with bacterial overgrowth in the distal small intestine. Intravenous administration of dextran 70 prevented bacterial overgrowth and translocation, at least in part, by maintaining gut vascular endothelial barrier integrity PMID- 9395791 TI - High doses of hydroxyethyl starch and human albumin have similar effects on monocyte function and oncotic pressure. AB - The accumulation of hydroxyethyl starches (HES) in monocytes/macrophages has raised concern over their potential detrimental effects on host defences. We assessed prospectively the function of circulating monocytes isolated from patients treated with plasma exchange (PE) using HES. The study was carried out in the medical intensive care unit of a university hospital. Eight patients underwent PE for neurological disorders. Each patient underwent three PEs, 48 h apart. The total exchange volume was 4 L per PE. Only 4% human albumin was used for the first PE. In the second and third PEs, the plasma substitute was 2 L of HES (200,000/6%/0.62) and 2 L of albumin. Mononuclear cells were collected before and immediately after each PE and 48 h after the last PE. They were placed in suspension culture and incubated with lipopolysaccharide (LPS). Monocyte function was assessed in terms of procoagulant activity (PCA) and tumour necrosis factor alpha (TNF-alpha) production. LPS-stimulated PCA increased after the first PE (P < 0.05). Stimulated TNF-alpha production increased, but not significantly so. Similar effects were observed after the second and third PE (P < 0.05 for stimulated TNF-alpha). Values 48 h after the last PE were similar to those obtained before the second PE, suggesting that repeated infusions of HES had no detrimental effect on monocyte function. Furthermore, plasma oncotic pressure was preserved after PE with HES. These results support the partial replacement of costly human albumin with HES during repetitive PE, and suggest that HES might be a safe plasma expander in septic patients. PMID- 9395792 TI - Specific activation of AP-1 but not Stat3 in regenerating liver in mice. AB - Liver regeneration following partial hepatectomy is regulated by hepatotrophic factors whose precise roles are still elusive. In cell culture studies, some of them have been shown to activate members of the family of the signal transducers and activators of transcription (Stat). To test this contention in vivo, nuclear extracts were isolated from livers of partially hepatectomized and sham-operated mice killed at 30 different time points between zero and 108 h after surgery. Stat3 DNA binding is rapidly induced after surgery in both partially hepatectomized and sham-operated mice. Maximum activation of Stat3 is achieved 4 6 h after resection, and elevated Stat3 activation is detected as late as 60 h after surgery in both groups. Activated Stat5 is found sporadically in both sham operated and resected mice but appears to be absent in the first 12 h after partial hepatectomy. Neither Stat1, Stat2, Stat4 nor Stat6 is induced during the time of observation. In contrast, AP-1 DNA binding activity is specifically induced in regenerating mouse liver. PMID- 9395793 TI - Rapid screening for amyloid-related variant forms of transthyretin is possible by electrospray ionization mass spectrometry. AB - We have used a new and rapid method to detect three variant forms of transthyretin (TTR): methionine for valine at position 30 (Met-30), serine for cysteine at position 6 (Ser-6) and methionine for leucine at position 111 (Met 111). By using an anti-transthyretin antibody and a centrifugal concentrator, transthyretin was isolated from plasma samples and analysed for variant forms by electrospray ionization mass spectrometry. Differentiation between homozygous and heterozygous transthyretin Met-30 and Met-111 posed no problems. However, a clear separation of transthyretin Ser-6 and Met-111 peaks in one patient with concordant Ser-6 and Met-111 mutations could not be achieved. The present method enables a quick and reliable detection of variant TTR. It can be used to screen families or small populations for abnormal TTR. Knowledge of TTR polymorphism and the variable expression of different amyloidogenic mutations should be taken into consideration when applying the methods in clinical practice. PMID- 9395794 TI - ATP-sensitive potassium channels mediate vasodilatation produced by calcitonin gene-related peptide in human internal mammary but not gastroepiploic arteries. AB - The purpose of this study was to elucidate the mechanism of action of calcitonin gene-related peptide-induced vasodilatation of human gastroepiploic and internal mammary arteries. Calcitonin gene-related peptide (0.1-100 nmol L-1) elicited relaxations of preconstricted vessels, with a significantly greater effect in the gastroepiploic artery (P < 0.05). This effect was independent of endothelium derived vasodilating substances. The response of the internal mammary artery but not the gastroepiploic artery to calcitonin gene-related peptide was attenuated by glybenclamide (1.0 mumol L-1) (P < 0.05). In vitro autoradiography indicated that [125I]-calcitonin gene-related peptide bound to the tunica media but not the endothelial cells in both types of artery, with a significantly higher degree of binding in the gastroepiploic artery. It is concluded that calcitonin gene related peptide acts directly on vascular smooth muscle via specific binding sites to induce vasodilatation. In addition, KATP channels are involved in the action of calcitonin gene-related peptide in the internal mammary artery but not in the gastroepiploic artery. PMID- 9395795 TI - Contribution of nitric oxide to exercise-induced changes in healthy volunteers: effects of acute exercise and long-term physical training. AB - Endothelium plays a central role in the regulation of regional blood flow through the release of certain vasoactive substances. We conducted this study to test whether an increase in the production of nitric oxide (NO) metabolites, atrial natriuretic peptide (ANP) and plasma and intraplatelet cyclic guanosine 3':5' monophosphate (cGMP) is involved in the adaptation to chronic exercise in physically trained people and in the vasodilatation induced by acute physical exercise. We studied one group of 10 trained athletes and another group of 10 untrained people. We measured plasma levels of nitrites, nitrates and cGMP and intraplatelet levels of cGMP, as an indicator of intracellular guanylate cyclase activity, and ANP before and after a maximal treadmill test. Resting cardiac rate (CR) and systolic blood pressure (SBP) were lower in the athlete group than in the control group (73.8 +/- 3.6 vs. 92 +/- 5.9; P < 0.02 and 110 +/- 2.58 vs. 118 +/- 3.27; P < 0.02 respectively). SBP did not show differences between groups after the exercise test. Diastolic blood pressure (DBP) at rest was lower in the athlete group (71 +/- 1.79 vs. 80.5 +/- 3.53; P < 0.03) and the decrease after maximal exercise was more pronounced in this group (64 +/- 2.67 vs. 74.5 +/- 3.2; P < 0.02). Basal plasma nitrites were 4.9 +/- 0.8 in the athlete group and 1.9 +/ 0.3 in the control group (P < 0.05). After exercise, test differences between groups remained (P < 0.05). Nitrates were significantly higher in the group of athletes and did not show exercise-related changes. Plasma levels of cGMP and ANP increased in both groups after the treadmill test, with no differences between groups. Among the athletes, cGMP increased from 1.11 +/- 0.1 to 2.6 +/- 0.4 (P < 0.001), whereas in the untrained group plasma cGMP rose from 1.14 +/- 0.09 to 1.86 +/- 0.2 (P < 0.01). There was a significant correlation between the increases in plasma cGMP and the atrial natriuretic peptide in both groups (r = 0.91, P < 0.0002, for athletes; and r= 0.68, P < 0.04, for control group). The intraplatelet concentration of cGMP did not show differences between groups and did not change after exercise. In conclusion, we have found increased basal levels of plasma nitrite and nitrate in trained subjects. Exercise does not produce differences in the increments of these metabolites. Therefore, we speculate the release of nitric oxide is not augmented by exercise in trained athletes. PMID- 9395796 TI - The impact of electronic publication and the E-journal on quality and the peer review process. PMID- 9395797 TI - Penetrating keratoplasty in the Singapore National Eye Centre and donor cornea acquisition in the Singapore Eye Bank. AB - We analyzed all penetrating keratoplasties performed in the Singapore National Eye Centre from 1 January 1991 to 31 December 1995, using records of the Singapore Eye Bank Registry, evaluating the indications, complications, causes of graft failure, visual outcome and graft survival rate. We also looked into donor cornea acquisition in the Singapore Eye Bank and its influence on the development of corneal transplantation in the Singapore National Eye Centre. A total of 327 penetrating keratoplasties were performed during the 5-year period. Bullous keratopathy was an indication in 26.3% of cases. Of these, aphakic bullous keratopathy accounted for 11.6% of all cases, while pseudophakic bullous keratopathy accounted for 11.3%. Other indications were regrafts (11.9%), corneal dystrophies (10.4%), traumatic corneal scarring (10.1%) and keratoconus (9.8%). Graft rejection was a complication in 20% of all cases. Of these, 40.9% led to graft failure. Other major complications were raised intraocular pressure (18%), epithelium-related problems (7.3%), wound dehiscence (4.3%), cataract (3.3%) and microbial keratitis (3.1%). The main causes of graft failure were graft rejection (8.2%), endothelial failure (2.4%), infection (2.4%) and glaucoma (2.1%). Of the 327 grafts, 40.3% achieved best corrected visual acuity of 6/12 or better; 70.8% achieved vision of 6/24 or better. The overall graft survival rate was 82.3% after a mean follow-up period of 2 years. Donor corneas for the penetrating keratoplasties were obtained from foreign eye banks as well as locally, with the local donation rate steadily increasing from 1991 to 1996, with the establishment of proper eye banking facilities and the Singapore Eye Bank. These results show that the indications and outcome of penetrating keratoplasty in the Singapore National Eye Centre are similar and comparable to that of other centres with established corneal grafting programmes. The establishment of the Singapore Eye Bank has ensured the proper co-ordination of acquisition of donor material which has been vital to the development of corneal transplantation in the Singapore National Eye Centre. PMID- 9395798 TI - Photorefractive keratectomy for the treatment of compound myopic astigmatism using the ablatable mask. AB - Eight eyes of 8 patients with compound myopic astigmatism were treated with excimer laser photorefractive keratectomy (PRK) using a hand-held ablatable mask in conjunction with the Summit excimer laser. The attempted correction ranged from -1.25 to -400 dioptres (D) of astigmatism and 0 to -8.00 D of myopia. All eyes had attained at least 6 months of postoperative follow-up. Five of the 8 eyes achieved an unaided visual acuity of 6/12 or better. Postoperative refractions ranged from -0.50 to -3.50 D of refractive cylinder and from +0.50 to -3.75 D of spherical error. Decentration of the ablation zone was encountered in 3 eyes due to shifts in patients' fixation. Technical difficulty with the use of the hand-held ablatable mask limited the widespread application of this procedure and it has now been superseded by newer excimer laser systems which can correct astigmatism without having to employ a mask. Despite this, because of the theoretical ability of the mask to correct any form of refractive error, the concept of the mask shape transfer process will remain as a potential alternative in refractive surgery, especially for correction of hyperopia and hyperopic astigmatism. PMID- 9395799 TI - Hydroxyapatite orbital implants--our local experience. AB - Hydroxyapatite orbital implants were first introduced by Arthur Perry in 1985 and approved by the Food and Drug Administration for general use in 1989. Since its inception, it has become a popular orbital implant as it combines biocompatibility with improved motility. At the Singapore National Eye Centre, twenty consecutive patients underwent hydroxyapatite implantation from June 1993 to June 1996. Seventeen patients who had a follow-up time of more than 6 months were analysed retrospectively. Sixteen had enucleation while one had evisceration performed. The implant size used was either 18 or 20 mm. Seven primary and 10 secondary hydroxyapatite implants were performed in 17 patients ranging in age from 20 to 65 years. Six consisted of coralline hydroxyapatite while the remaining 11 were made of artificial hydroxyapatite. In the follow-up time of 6 to 16 months (mean 10.2 months), there have been no cases of infection, extrusion or migration. Two patients who had implant exposure were managed surgically with fascia lata grafts and have healed well. It appears that hydroxyapatite is the implant of choice for an anophthalmic socket. It becomes incorporated by the patients own tissues and once vascularised, it is less likely to migrate or extrude. The complication rate is low and implant exposure, if it occurs, can be easily managed. PMID- 9395800 TI - Acute orbital cellulitis--a review of 17 cases. AB - This is a retrospective study of 17 patients who were admitted for orbital cellulitis between August 1989 and February 1994. The epidemiology, possible source of infection, the causative organisms and effectiveness of treatment were reviewed. The results showed that one-third of the cases occurred in the first decade of life. Paranasal sinusitis was the main source of infection in 11 (65%) patients. Thirteen patients (76.5%) developed orbital and periocular abscesses requiring surgical drainage. Two out of 17 eyes lost vision despite intensive treatment. Orbital cellulitis is a blinding ocular emergency associated with high morbidity. Immediate treatment is necessary to avoid devastating complications such as optic nerve compression, panophthalmitis or intracranial spread of infections. Cases with abscess formation required early surgical drainage. Combined orbito-otorhinolaryngologic approach is recommended for drainage of orbital abscess with associated paranasal sinus infection. PMID- 9395801 TI - Detection of low numbers of neuroblastoma cells in vitro. AB - Neuroblastoma is a tumour derived from the sympathoadrenal progenitors of the neural crest. It is one of the most malignant solid tumours in childhood with an annual incidence of 9.4 per 10(6) children under 15 years of age. Recent studies suggest that immunocytological detection of neuroblastoma cells in bone marrow and circulating neuroblasts during treatment may predict clinical outcome and correlate with tumour relapse. The present methods of diagnosing metastasis in neuroblastoma include histological, biochemical and immunohistological analysis. Morphological distinction between tumour cells and primitive lymphoblasts in bone marrow is often difficult, and these methods may also not always be sensitive enough for early detection of the residual and minimally circulating tumor cells. A sensitive assay for detection of such residual cells using two tissue-specific markers, NFM and SYN, by reverse transcriptase-polymerase chain reaction (RT-PCR) is reported here. Analysis of the specificity of this assay in three neuroblastoma cell lines, namely IMR 32, SK-N-SH and SY5Y showed positive expression while control peripheral blood mononuclear cells (HL 60) were negative. In reconstituted cell spiking tests, this method has the ability to detect 1-10(3) neuroblastoma cell in 10(7) normal peripheral blood mononuclear cells (PBMC), as shown by serial dilution and limiting dilution. The NFM marker was found to be a more sensitive marker. The specificity and sensitivity of this technique makes it suitable for future application in detection of minimally disseminated tumour cells in neuroblastoma patients. PMID- 9395802 TI - Group B streptococcus: maternal carriage rate and early neonatal septicaemia. AB - Between January 1984 and December 1994, 30 cases of early neonatal group B streptococcus (GBS) septicaemia were managed in the Neonatal Unit, University Hospital, Kuala Lumpur. Two neonates were outborn and 28 were inborn, giving an average annual incidence of neonatal GBS septicaemia of 0.4/1000 livebirths among inborn babies. In a separate survey over a three-month period, GBS genital carriage rate among 196 parturients was found to be 9.7%. Of the infants with GBS septicaemia, the mean gestational age was 37.5 +/- 3.8 weeks and the mean birthweight was 2540 +/- 716 g. Twelve (40%) were preterm infants and 14 (47%) were low birthweight infants. Male and female infants were almost equally affected. Prolonged rupture of membranes and maternal pyrexia accounted for only 5 (17%) and 3 (10%) of the cases respectively. Twenty-four (80%) neonates had onset of symptoms within 6 hours of life and respiratory symptoms were observed in 24 (80%) of the cases, while meningitis was uncommon. Six (20%) neonates died. Preterm and low birthweight infants had higher mortality than their term counterparts: 42% versus 6% and 36% versus 6% respectively. Of those who died, 4 (67%) required respiratory support right from birth and the mean time of onset of symptoms was 4 hours (range 0 to 21 hours) and the duration of survival was only 28.8 hours (range 12 to 38 hours). As the incidence of neonatal GBS septicaemia was low, mass screening and chemoprophylaxis for GBS were not recommended. All the GBS isolates were sensitive to penicillin and ampicillin, thus one of these antibiotics should be included in the antimicrobial therapy of septic neonates. PMID- 9395803 TI - Preoperative versus postoperative pethidine for extraction of impacted third molars. AB - We have studied the pre-emptive analgesic effects of pethidine by comparing its analgesic effects given before or immediately after operation in a randomized, double-blind study of 40 patients undergoing removal of bilateral impacted third molars under general anaesthesia. Group 1 patients received pethidine 50 mg as a 1 ml injection 1 to 2 hours before operation and normal saline 1 ml intramuscularly immediately after surgery. Group 2 patients received normal saline 1 ml intramuscularly before operation and pethidine 50 mg as a 1 ml injection immediately after surgery. Outcome measures included perception of pain on a visual analogue scale (VAS), the number of patients who required postoperative pethidine, time to first postoperative pethidine injection and total dose of pethidine given. Four patients in group 1 compared to 8 in group 2 required postoperative pethidine but this was not statistically significant. The VAS scores, time to first postoperative pethidine injection and total dose of pethidine also did not differ significantly between the 2 groups. We concluded that preoperative administration of pethidine intramuscularly did not confer additional analgesic effects compared with a similar dose given after surgery. PMID- 9395804 TI - Preoperative prediction of intra and postoperative red blood cell transfusion in surgical patients. AB - Anaesthetists were asked to predict the need for intra and postoperative red blood cell transfusion in 1706 patients before surgery. Each prediction was made using only the individual patient's medical history and physical examination, the results of routine preoperative laboratory investigations, and knowledge of the proposed surgical procedure. Only 159 patients (9.3%) received red blood cell transfusion. The sensitivity and specificity of this preoperative prediction were 85.5% and 96.6% respectively, whereas the positive and negative predictive values were 72.3% and 98.5% respectively. Using a stepwise logistic regression model, preoperative haemoglobin concentration, Surgical Table of the procedure, and age of patients were found to significantly determine the need for intra and postoperative red blood cell transfusion. It is recommended that type and screen should replace group and crossmatch procedures in surgical patients where no intra and postoperative red blood cell transfusion is predicted as necessary. PMID- 9395805 TI - Thyroid stimulating hormone receptor antibody levels in Singaporean patients with autoimmune thyroid disease. AB - Stimulating thyrotrophin receptor antibodies (TRAbs) have been identified as the antibodies responsible for the pathogenesis of Graves' disease (GD) while blocking TRAbs have been implicated as the cause of hypothyroidism in some patients with chronic lymphocytic thyroiditis (CLT). TRAb positivity in patients with other thyroid disorders such as silent thyroiditis, toxic multinodular goitre and subacute thyroiditis has been reported but the role of TRAb in these disorders is unclear. A study was carried out to determine the prevalence of TRAb positivity in Singaporean patients with a spectrum of thyroid diseases. TRAb levels were measured in 181 patients with GD, 54 patients with CLT (37 goitrous and 17 agoitrous), 16 patients with thyroid nodules, 11 patients with subacute thyroiditis, 1 patient with hyperthyroidism due to a human chorionic gonadotrophin (HCG)-secreting tumour, 2 patients with thyroid stimulating hormone secreting tumours and 2 patients with amiodarone-induced dysthyroidism. Using a cut-off of 10.0 U/L, TRAb levels were found to be positive in 79.0% of GD patients, 9.2% of CLT patients (euthyroid and hypothyroid) and no patients with other thyroid disorders. TRAb was a more sensitive marker of GD than anti thyroglobulin antibodies (53.2%) but not anti-microsomal (78.3%) antibodies. TRAb levels > 10.0 U/L appear to be highly specific for autoimmune thyroid disease. PMID- 9395806 TI - Usefulness of bacteriologic cultures in choice of antibiotics in patients with chemotherapy-induced neutropenic sepsis. AB - Neutropenic sepsis is a potential problem in cancer patients undergoing cytotoxic chemotherapy. Septic work-up including cultures from various sites is routinely done for these patients. To assess the usefulness of these cultures, a retrospective review of all patients admitted for neutropenic sepsis in the period from June 1994 to August 1995 was conducted. All patients included in the study had solid tumours which were being treated at our institution during the study period. All had fever and documented neutrophil count of < 1 x 10(9)/l on at least one occasion. There was a total of 41 patients with 52 episodes of neutropenic sepsis. Of the 52 episodes, there were positive cultures in 14 (27%) episodes, including 7 from blood, 2 from urine and 5 from skin. In the bacteriologic cultures, gram-negative bacteria were isolated in 11 episodes and gram-positive bacteria in 5 episodes (2 episodes had both gram-negative and gram positive bacteria isolated, and 1 episode had two gram-negative bacteria). Majority of the patients (96%) were treated with a third generation cephalosporin with/without an aminoglycoside. This empirical treatment was effective with resolution of fever in 39 (75%). Thirteen (25%) had change of antibiotics because of deteriorating clinical state or drug resistance. Nine patients with unabated sepsis had bacteriological cultures which grew organisms resistant to the empirical antibiotics. Eight responded to the change of antibiotics. One patient with pseudomonas bacteraemia failed to respond to empirical treatment with ceftriaxone and died before antibiotics could be changed. Of the patients with positive cultures, the results of drug sensitivity made a difference to treatment. None of the patients with negative cultures died from sepsis. It appears that even though the rate of positive culture is low (27%), it is still useful as a guide when change of antibiotics is required. PMID- 9395807 TI - Profile of patients seeking psychiatric treatment from the adult public mental health services in Singapore. AB - This study examined the characteristics of patients seeking help from public mental health services and changes in the utilisation of the services over time. The sample included 198 consecutive new referrals above the age of 15 years to the adult psychiatric services provided by Woodbridge Hospital and the Institute of Mental Health from 1 January 1995 and followed up over a one-year period. The most common reason for seeking psychiatric treatment was for abnormal behaviour (26%) and the most common diagnostic group was schizophrenia/paranoid disorder (21.1%). The medical sector was the most common source of referral (56%). The primary health care services referred significantly more patients with anxiety disorders while the police and relatives/friends referred significantly more patients with schizophrenia/paranoid disorder. About one-third of the sample made their first contact with the services at the 24-hour Admission Room. One third received inpatient treatment after their first consultation and those with schizophrenia/paranoid disorders were more likely to be admitted (P < 0.05). At the end of one year, 23.2% remained in the services. There were significantly more elderly patients and those suffering from schizophrenia/paranoid disorder maintaining contact with the services after one year. PMID- 9395808 TI - Hypercholesterolaemia and its treatment in Singapore with implications for screening. AB - The National University of Singapore Heart Study is a cross-sectional survey of the whole of Singapore from 1993 to 1995 with a random sample of 961 persons aged 30 to 69 years (response rate 71.2%). Serum lipids were measured enzymatically on an autoanalyser (Ektachem, kodak). Hypercholesterolaemia was diagnosed if the person was on lipid lowering drugs or had a serum cholesterol > or = 6.5 mmol/L (250 mg/dL). Results for the 40 to 64 age group are presented. The proportion and number in Singapore with hypercholesterolaemia were high, 23.7% and 186,544 persons respectively. However, they declined with the additional requirement of coronary heart disease (CHD) or risk factors (i.e. cigarette smoking, hypertension or diabetes mellitus), and were small if only hypercholesterolaemic persons with CHD were included, 1.6% and 13,126 persons respectively. For both genders, the proportions on drugs increased with the added options of CHD or risk factors, so that for persons with hypercholesterolaemia and CHD the proportion on drugs was very high i.e. 84.0%. However, 11.2% of persons with only hypercholesterolaemia (i.e. without CHD or other risk factors) had been prescribed drugs. The main conclusions were:1) The use of different criteria for the screening and treatment of hypercholesterolaemia would involve large differences in the number of Singaporeans on lipid lowering drugs and this should be considered when deciding policy and 2) Medical practitioners currently to some extent do consider the presence of CHD or other risk factors when prescribing lipid lowering drugs for hypercholesterolaemia. PMID- 9395809 TI - Predilection for supracondylar fractures in children with cubital recurvatum. AB - It has been a common observation that children with supracondylar fracture of the humerus tend to demonstrate hyperextensibility of their elbow joints. In this study, we measured and compared the degree of hyperextension of the uninjured elbow of children less than sixteen years old who sustained either a supracondylar or a distal radial fracture. We concluded that children with higher degree of hyperextension in the elbow are more likely to sustain a supracondylar fracture than a distal radius fracture during a fall on the outstretched hand. PMID- 9395810 TI - An epidemiological study of developmental dysplasia of the hip in infants in Singapore. AB - The incidence of congenital dislocation of the hip (CDH) in Singapore and Malaysia has been reported as being lower than in the West. In our hospital, we have seen an increasing number of congenital hip dislocation as well as dysplastic hips. We undertook a prospective study from December 1989 to December 1994 of 20,000 live births. The neonates were all screened by a consultant neonatologist and the findings were confirmed by a consultant paediatric orthopaedic surgeon. All babies had plain X-rays at 3 months and an acetabular index (AI) of 30 degrees or more was considered dysplastic. All babies with positive signs were followed up for 1 year and again had radiographs taken at 1 year. Comparison of plain X-rays and ultrasound assessment in a subgroup of 130 neonates showed that 64% of patients with AI > 20 degrees had hip dysplasia by ultrasonographic (alpha angle < 60 degrees) The incidence of dysplastic hips was 16.8 per 1000 live births. The overall incidence of neonates with dislocated hips was 4.7 per 1000 live births. The Malays were most affected with an incidence of 5.4 per 1000 live births. The incidence of developmental dysplasia of the hip in Singapore is higher than previously reported, with the Malays having the highest incidence. A significant number of babies with clicking hips have radiological evidence of acetabular dysplasia (AI > 30 degrees). One-third of the babies' hips were still dysplastic at 1 year of age. A well-organised screening programme with experienced examiners has proved to be useful in making early and accurate clinical diagnosis. PMID- 9395811 TI - Uncemented total hip replacements using the Mecron acetabular cup--a prospective study. AB - The uncemented threaded cup gained some popularity over the past decade, firstly as a revision and then as a primary procedure. We elected to test the Mecron acetabular threaded cup system as a routine hip replacement. As there was no matched femoral component, we decided to use an uncemented femoral stem of the Ring system of the hip prosthesis. All patients were logged on to the study at the outset although this is not a controlled comparison with another system. Patients were assessed annually for up to 4-years at the time of reporting. There were 104 primary total hip replacements in 99 patients between 1987 and 1991. The mean follow-up for this report was 22.5 months (range 12 to 48 months). The mean age was 76.3 years (range 24 to 88 years) and the female to male sex ratio was 3.3:1. Within the study period, 5 hips required revision, all for acetabular loosening. Patients were assessed by the modified Harris score annually and 60.6% of the cases had an excellent or good short-term result. Poor results were observed in 18.3% of cases. Radiographic studies of the acetabular component were undertaken annually and measured. Migration and tilting of the acetabular cup was observed in all cases. Heterotropic bone formation occurred in 10 patients. As a result of these poor results, the prosthesis was abandoned in 1991. The cohort of patients continues to be followed. Failure of the Mecron acetabular cup in our series was mainly due to tilting and migration and not untwisting. PMID- 9395812 TI - Pattern of proteinuria in tubular injury and glomerular hyperfiltration. AB - Proteinuria is one of the bad prognostic indices in renal disease. This study compares the pattern of protein excretion in 10 patients with IgA nephropathy (IgAN), 10 patients with chronic glomerulonephritis approaching end-stage renal failure (ESRF) who still had proteinuria and 10 other patients with diabetic nephropathy (DN) with proteinuria but normal renal function. The pattern of proteinuria was analysed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), isoelectric focusing (IEF) and assayed for orosomucoid, alpha-1-microglobulin, retinol-binding protein, lysozyme, beta-2 microglobulin and N-acetyl-beta-D-glucosaminidase activity. Our data showed much similarity in the pattern of proteinuria between the DN and ESRF groups but significant differences with the IgAN group. The pattern of proteinuria in the IgAN group reflects glomerulonephritis whereas the similar pattern between the ESRF and DN groups may reflect hyperfiltration as well as tubular injury. PMID- 9395813 TI - Refractory seizures in a young army cohort. AB - This survey covered male Singapore citizens born in 1974 who were medically screened at the age of 18 years before enlistment for compulsory military service. Suspected epileptics were referred to government hospitals for further management. Out of 20,542 men, there were 121 epileptics, giving a cumulative incidence of 5 per 1000 by age 18 years. We had information on 106 (87%) of these individuals and were able to interview them and review their hospital records. Seventy-three of the 106 (69%) epileptics had generalised seizures while 14 (13%) had refractory seizures. There was no statistically significant racial bias amongst these epileptics. Unprovoked afebrile seizures occurred early in these patients, half of whom had seizures onset before 7 years of age. Nine refractory epileptics had a history of febrile seizures, 4 of which were complex febrile seizures. Magnetic resonance imaging identified mesial temporal sclerosis in 2 patients and a hypothalamic lesion in 1 patient. Computed tomographic scans revealed focal cortical atrophy in 2 patients. Nine other patients had normal imaging studies. Nine out of 14 (64%) patients with refractory epilepsy had partial seizures; 4 (29%) had generalised seizures and 1 (7%) was unclassified. This is in contrast to the distribution of the entire cohort of epileptics studied. Two out of 9 patients with refractory partial seizures (gelastic epilepsy and mesial temporal sclerosis) had undergone surgery while 6 of the other 7 patients refused to consider surgery. PMID- 9395814 TI - Surgical excision of intracranial arteriovenous malformations after preoperative embolisation with N-butylcyanoacrylate. AB - The aim of this study was to determine the usefulness of preoperative embolisation of intracranial arteriovenous malformations (AVMs) with N Butylcyanoacrylate (NBCA) since the introduction of this interventional neuroradiology technique in March 1994 at Tan Tock Seng Hospital, Singapore. Twenty-one patients who underwent complete surgical excision of their AVMs (proven on postoperative angiograms) were studied. Eight patients had preoperative embolisation with NBCA prior to surgical excision of their AVMs. Thirteen patients had excision of their AVMs without preoperative embolisation and these were used as the control group. The parameters studied were the patient's AVM characteristics, the amount of blood loss and the length of operative time. Statistically significant reduction in blood loss occurred in Spetzler and Martin Grade 3 and 4 AVMs but not in Grade 1 and 2 AVMs undergoing preoperative embolisation with NBCA. Operative time was reduced in Grade 3 and 4 AVMs but not in Grade 1 and 2 AVMs, although this was not statistically significant. Preoperative embolisation of AVMs is hence a useful and important adjunct in the management of patients with Grade 3 and 4 AVMs of the brain undergoing conventional open microneurosurgery. PMID- 9395815 TI - Glycoprotein IIb/IIIa platelet receptor inhibitors: a new dimension in cardiology. AB - Platelets play a key role in the pathogenesis of atherosclerosis, acute coronary syndromes and ischaemic complications following percutaneous coronary intervention. More recently, the platelet glycoprotein (GP) IIb/IIIa receptor has been identified as the pivotal mediator of platelet aggregation, leading to thrombus formation. This has led to the emergence of a novel class of potent antiplatelet agent, the GP IIb/IIIa receptor antagonists. These agents show great promise in reducing ischaemic complications of coronary angioplasty and acute coronary syndromes. This review summarizes the current state of knowledge of the biology of GP IIb/IIIa receptor, the different classes of GP IIb/IIIa receptor antagonists, the results of the various trials, and their impact on current clinical practice. PMID- 9395816 TI - The epidemiology of obesity: a review. AB - Obesity is a major public health problem worldwide contributing to increased morbidity and mortality. There are several indices of obesity which include body mass index, waist-hip circumference ratio and skinfold thickness. The prevalence of obesity varies from 7% in France to 32.8% in Brazil. However, the comparison of obesity across different countries is difficult as there are different age structures of the population, measurement techniques are not the same and surveys may not be population-based. Trends in developed and developing countries suggest that the rates of obesity are increasing. This rising trend may be contributed by factors such as low levels of physical activity, high calorie intake and long hours of watching television. Cohort and cross-sectional studies have shown that obesity may be linked to an increased risk of coronary heart disease, hypertension, diabetes mellitus and gallstones. There is also a positive association between obesity and cancer. The mortality of obese adults, adolescents and children is higher than that of the general population. Multi prolonged intervention strategies are needed to prevent obesity and its associated complications. PMID- 9395817 TI - Vocal cord dysfunction: two case reports. AB - Vocal cord dysfunction is an uncommon condition characterised by adduction of vocal cords that can masquerade as or coexists with bronchial asthma. The glottic dysfunction is due to a functional (non-organic) cause. If unrecognised, incorrect diagnosis may result in patients being unnecessarily treated as refractory or severe asthma with high doses of corticosteroid. This may result in unwarranted steroid toxicity. Clues that should raise clinical suspicion to the diagnosis of vocal cord dysfunction include lack of response to bronchodilators, poor reproducibility of spirometric indices due to inconsistent effort and truncation of the inspiratory limb of the flow-volume loop. Definitive diagnosis is made by direct visualisation of the vocal cords during an attack. We report two patients with vocal cord dysfunction and review the literature on this disorder. PMID- 9395818 TI - A case of fluid embolism from transcervical endometrial resection. AB - Hysteroscopic transcervical endometrial resection (TCER) using a urological resectoscope offers an alternative to hysterectomy for some women with menorrhagia. The advantages of this technique are a shorter operating time and a rapid recovery and discharge. However, this procedure is not without risk of complications and one of them is related to dilutional hyponatraemia from absorption of the irrigation solution. This report describes a patient who experienced such a problem. PMID- 9395819 TI - Familial aplasia cutis congenita of the scalp: a case report and review. AB - We report three cases of aplasia cutis congenita of the scalp affecting two siblings and their mother, suggesting group 1, or autosomal dominant aplasia cutis congenita not associated with multiple abnormalities. A review of the clinical features and literature concerning this heterogenous and rare condition is presented. PMID- 9395820 TI - Familial papillary thyroid cancer: a case report. AB - Familial occurrence of medullary thyroid cancer is well known in families as an isolated malignancy or in association with multiple endocrine neoplasia syndrome type II. Conversely, papillary thyroid cancer almost always presents sporadically except for reports of familial clustering in individuals with radiation exposure, inherited syndromes of colonic polyposis or multiple harmatomas, and rarely in monozygotic twins. We report a case of papillary thyroid cancer diagnosed incidentally in a 53-year-old woman who underwent surgery for excision of an adenomatous nodule. It was noted that her mother suffered from a similar thyroid malignancy some 33 years ago, and several of her maternal relatives had either Graves' disease or hypothyroidism. The possible existence of this familial entity and its likely genetic basis is discussed. PMID- 9395821 TI - De novo interstitial deletion of chromosome 1p with absent corpus callosum--a case report. AB - Deletion of short arm of chromosome 1 is a rare clinical entity and there are no clearly defined phenotype. We report a case of deletion of the short arm of chromosome 1, which is believed to be the first case among the Chinese population. This baby was also found to have some Robinow Syndrome-like features as well as absent corpus callosum which have never been reported in deletion of chromosome 1p. PMID- 9395822 TI - Histoplasmosis in the head and neck. AB - Histoplasmosis is a fungal infection which when disseminated can be potentially fatal, especially in the immunocompromised patient. Disseminated histoplasmosis can present with orolaryngeal manifestations which often mimic carcinoma or tuberculosis. A high index of suspicion and diagnosis with biopsy is needed. Two recent cases are presented and the literature reviewed. As the number of immunocompromised patients increases, more cases of histoplasmosis in the head and neck will be encountered. Early treatment can reduce morbidity and mortality. PMID- 9395823 TI - Popliteal artery aneurysm: two case reports of acute lower limb ischaemia. AB - Two cases of thrombosed popliteal artery aneurysm causing acute ischaemia were described. This report emphasises the importance of early diagnosis and surgical intervention of this condition. We then present a brief review of clinical presentation and current management of popliteal artery aneurysm. Popliteal artery aneurysm commonly first present as acute lower limb ischaemia secondary to thrombo-embolism. Failure to recognise this limb-threatening condition can lead to delay in treatment and even limb loss. It is therefore important to increase awareness of this condition. Popliteal artery aneurysm warrants early surgical intervention, even in an asymptomatic stage because of its potential for serious complications. PMID- 9395824 TI - Cholesterol and atherosclerosis: a contemporary perspective. AB - Monumental advances in the field of lipid metabolism and its relationship to atherosclerotic cardiovascular disease have been achieved during the last half century. Epidemiologic studies have defined lipid disorders as highly significant independent risk factors for coronary heart disease, along with diabetes mellitus, hypertension and smoking. Primary and secondary prevention studies including the Coronary Primary Prevention Trial, Helsinki Heart Study, and the Coronary Drug Project have shown that lowering the atherogenic low density lipoproteins (LDL) and very low density lipoproteins (VLDL) whilst raising the high density lipoproteins (HDL) significantly decreases the risk for coronary disease. Striking evidence that aggressive therapy (to sharply lower LDL and raise HDL with newer drugs) prevents progression and induces regression of coronary narrowing has been obtained in numerous recent studies using quantitative coronary arteriography. An interesting and unexpected lesson learned from these arteriographic studies was that a highly significant reduction within months in several studies in coronary events was out of proportion to improvements in luminal narrowing. Recently, three major clinical trials to assess the effects of cholesterol reduction by the newly discovered HMG CoA reductase inhibitors (statins) have been published. Pravastatin significantly reduced coronary events in hypercholesterolemic patients [mean LDL-Chol. = 5.0 mM/L (192 mg/dl)] without a history of myocardial infarction. In a secondary prevention study, simvastatin also reduced coronary complications in hypercholesterolemic patients [mean LDL-Chol. = 4.9 mM/L (190 mg/dl)] with pre existing coronary disease. Very recently, pravastatin treatment significantly reduced coronary events and stroke in patients with a history of myocardial infarction and average cholesterol levels [mean LDL-Chol. = 3.6 mM/L (139 mg/dl)], representing the majority of patients with coronary disease. In all these studies, reduction in cardiovascular events was approximately one-third. In subgroup analyses, men, women, elderly, smokers and hypertensives benefited from cholesterol lowering. There was no significant increase in non-cardiovascular causes of death. In the United States of America, the National Cholesterol Education Program (NCEP) Adult Treatment Panel, representing major health organizations, developed national guidelines on the detection, evaluation and treatment of high blood cholesterol in adults. In a given patient, the Panel recognizes the importance of weighing all cardiovascular disease risk factors including age (men > 45 years, postmenopausal women), family history of premature coronary disease, smoking, hypertension, diabetes and HDL-Cholesterol (< 35 mg/dl) in determining how aggressive therapy should be. The patient with manifest coronary heart disease (CHD) is given a special position as such patients are at highest risk for recurrent events. Major goals of therapy are to lower the LDL Cholesterol to 2.6 mM/L (< 100 mg/dl) in the CHD patient. In non-CHD patients with two or more risk factors, the LDL-Cholesterol goal is 3.4 mM/L (130 mg/dl). In those with fewer risk factors, the goal is 4.2 mM/L (160 mg/dl). These guidelines should be modified as appropriate for Singapore. Patients with elevated triglycerides usually have low HDL-Cholesterol levels and often represent a heterogeneous group who may have other concurrent abnormalities including the presence of small dense LDL, insulin resistance, hypertension, obesity, overt diabetes and combined hyperlipidemia. Such patients merit individualized treatment. The prevalence of this syndrome may be more common in Singapore and requires further investigation. Current therapeutic guidelines emphasize the need for weight loss and dietary restriction of total and especially saturated fat (< 7% to 10% total calories), cholesterol (< 200 to 300 mg/day), and exercise. (ABSTRACT TRUNCATED) PMID- 9395825 TI - Joint lectureship of the Royal College of Surgeons of Edinburgh and the Academy of Medicine, Singapore--accurate long-term documentation in surgical audit and evaluation of outcomes. AB - The purpose of a surgical audit is to allow a critical assessment of patient care, carried out among peers and in an atmosphere of learning, open discussion, free of recriminations. The results of a surgical unit will hopefully lead to improvement in the quality of care of our patients by reducing the preventable causes of mortality and morbidity. Data from surgical audit can also be used to monitor the teaching programme in a department. It may even allow us to study the cost effectiveness of certain procedures which is obviously very important in this era of diminishing resources and cost containment. PMID- 9395826 TI - Knowledge and beliefs about common eye diseases. PMID- 9395827 TI - Intraocular lens materials and styles: a review. AB - Biomaterial science has lead to the development of a variety of foldable intraocular lens (IOL) biomaterials. This literature review examines these lenses from both a basic science and a clinical perspective. By most parameters, hydrogel, soft acrylic and silicone IOL are better than polymethylmethacrylate (PMMA) lenses. Plate haptic silicone IOL have the lowest incidence of cystoid macula oedema and posterior capsule opacification, but these lenses require an intact anterior capsularhexis and posterior capsule. Yttrium aluminium garnet (YAG) laser capsulotomy must be delayed at least 3 months to avoid posterior lens dislocation. Silicone has the lowest threshold for YAG laser damage of all IOL materials and also adheres irreversibly to silicone oil with subsequent optical impairment. Three piece silicone IOL with polypropylene haptics have a higher incidence of decentration, pigment adherence and capsule opacification compared with PMMA haptics. Hydrogel lenses are very biocompatible and resistant to YAG laser damage, but pigment adheres to the surface more readily than PMMA. Soft acrylic IOL unfold slowly, resulting in controlled insertion, but it is possible to crack the lens and some lenses develop glistenings due to water accumulation. There are significant socioeconomic implications to the large differences in posterior capsule opacification rates between the various biomaterials and the lens styles. PMID- 9395828 TI - Topical anaesthesia for cataract surgery. AB - First, do no harm. We believe that the analgesia provided by topical anaesthetic is adequate for small-incision cataract surgery and does not compromise the safety of the surgery. In addition, the lack of amaurosis is ideal for day-case surgery, which itself is increasingly popular. If preventable, why not prevented? The greatest attraction of topical anaesthesia is its complete absence of the complications described for injectional local anaesthetic techniques. We therefore recommend that our colleagues consider topical anaesthetic for patients undergoing small-incision cataract surgery under local anaesthesia. Our policy for the past 3 years has been to use only topical or general anaesthetics for cataract surgery. PMID- 9395829 TI - Retrospective study of ocular surface squamous neoplasia. AB - BACKGROUND: Ocular surface squamous neoplasia (OSSN) encompasses the conditions of simple dysplasia to carcinoma in situ to invasive squamous cell carcinoma. It has a high rate of recurrence after treatment and the potential to metastasize. The present retrospective study was aimed at further defining the characteristics and clinical course of OSSN. METHODS: With ethical approval, the records of all major pathology laboratories in Queensland were surveyed. Two hundred and eighty eight cases were identified: 155 dysplasia, 71 carcinomas in situ and 62 invasive squamous cell carcinoma. The records were analysed and an attempt was made to contact and re-examine the patients. RESULTS: Ocular surface squamous neoplasia occurs mainly in males (78.5%) with a mean age of 60.1 years (range 20-88 years). They present as irritation (40.1%) and are located usually at the limbus (87.8%). The majority of OSSN are treated by simple excision (87.5%), after which there is a high rate of recurrence (23.3%). The main predictors for recurrence include histological grade of the lesion, corneal location and larger size (> 2 mm). CONCLUSIONS: Management of OSSN requires adequate excision and careful follow up to monitor any recurrence. As with other ultraviolet light-related conditions, preventative measures must remain the key to disease control. PMID- 9395830 TI - Acute posterior multifocal placoid pigment epitheliopathy: a long-term study. AB - PURPOSE: To study the long-term visual outcome in patients with acute posterior multifocal placoid pigment epitheliopathy (APMPPE). METHODS: Fifteen patients (28 eyes) with APMPPE were reassessed at Westmead Hospital Eye Clinic (Westmead, NSW, Australia) up to 18 years (mean 6.4 years) after their initial presentation to one of the authors (PM) or to their private ophthalmologist. Stereo retinal photographs and visual acuities taken at the initial presentation were compared with those obtained at the most recent follow-up visit. RESULTS: The last recorded visual acuity was 6/6 or better in 16 eyes (57%), 6/9-6/18 in four eyes (14%), 6/24-6/60 in five eyes (18%) and worse than 6/60 in three eyes (11%). Nine patients (60%) were treated with oral prednisone during the acute episode with little therapeutic effect observed. Six patients (40%) described a prodromal flu like illness. CONCLUSION: This study suggests that the long-term visual outcome following an acute episode of APMPPE may not be as favourable as initially portrayed by Gass. It confirms other study findings suggesting that APMPPE may not be a benign self-limiting disease. Older age at onset and initial foveal involvement were associated with a worse visual outcome. PMID- 9395832 TI - A review of 24 cases of Mohs surgery and ophthalmic plastic reconstruction. AB - PURPOSE: Mohs surgery (micrographically controlled excision) has been advocated as an effective method of dealing with infiltrative periorbital skin tumours. It has been shown to have high rates of tumour clearance with minimal loss of normal tissue, thus making oculoplastic reconstruction easier and functional preservation better. The aim of the present study was to confirm this. Guidelines for the selection of patients for Mohs surgery are discussed. METHODS: We retrospectively reviewed 24 cases of primary (n = 18) and recurrent (n = 6) periorbital basal and squamous cell carcinomas managed by Mohs micrographic excision and plastic reconstruction who presented to the Royal Perth Hospital between 1992 and 1996. RESULTS: Our high rate of tumour clearance (100%) was similar to that of previous studies, although our follow-up period was only 14.6 months. The fact that 50% of our patients with lid involvement had an intact posterior lamella after Mohs excision correlates with the high level of normal tissue preservation. The low rate of postoperative symptomatic problems suggests good maintenance of function. The infiltrative nature of these tumours was highlighted by the substantial proportion of cases (37.5%) that had a much larger excision defect than what was expected prior to excision. CONCLUSIONS: Our analysis confirms that Mohs excision and subsequent oculoplastic reconstruction is an effective method to use when managing periorbital infiltrative skin tumours. PMID- 9395831 TI - Knowledge and beliefs about common eye diseases. AB - PURPOSE: To ascertain the level of knowledge of common causes of blindness in an adult Australian population and to relate this to use of eye care services. METHODS: A population-based study of common eye diseases in an urban population aged 49 years or older was conducted. The questions were concerned with the awareness and knowledge of and the ability to describe three common eye diseases, namely cataract, glaucoma and age-related macular degeneration (AMD). RESULTS: Awareness of cataract (98%) and glaucoma (93%) were high in this population, but awareness of AMD was low (20%). Among people who were aware of the target eye disease, only 29% showed some knowledge of glaucoma, 26% showed some knowledge of AMD and 20% showed some knowledge of cataract; this was also low in people who had previous eye treatment, such as cataract surgery. Knowledge was related to education level, occupational prestige and knowledge of other eye diseases. After excluding people with a previous eye disease diagnosis, those people who were aware and had some knowledge of eye disease accessed eyecare services more frequently. CONCLUSIONS: Knowledge of common eye diseases is generally lacking. Age-related macular degeneration is the leading cause of blindness in Australia, yet only 20% of the present study population had heard of it. As there are often no early symptoms for glaucoma, community awareness of this disease and the need for screening of people at risk may allow timely diagnosis and more effective therapy before advanced visual field loss has occurred. An informed public is more likely to present earlier with visual symptoms before irreversible visual loss has occurred and is more likely to comply better with recommended therapy. PMID- 9395833 TI - Georg Bartisch: Ophthalmodouleia, der Augendienst, 1583. A treatise on service of the eyes and a review of the chapter on strabismus. AB - In 1583, Georg Bartisch, oculist and cutter for bladder stones at the court of Duke August I of Saxony, published at his own expense a 273 page textbook of ophthalmology. It contained 91 wood cuts and, in the present volume, they are presented in brilliant colour as they were in the original books prepared for presentation. The book was one of the first medical treatises to be published in the German vernacular instead of traditional Latin. It has been translated into English and published in gothic type to simulate the original. Treatment of diseases of the eye by medicines or surgery are reported in great detail. It gives an account of ophthalmology at the time of the early Renaissance when enlightenment was beginning to overtake the darkness of the Middle Ages. PMID- 9395834 TI - Aeromonas sobria endophthalmitis. AB - BACKGROUND: Aeromonas sobria causes a rare Gram-negative bacterial water-borne infection. It has been found in waters of North Queensland and South-east Asia. Of all Aeromonas species, A. sobria is the most virulent and invasive and has been reported to cause soft tissue infection and corneal ulcer. METHODS: A 14 year-old Caucasian male from North Queensland presented following a penetrating eye injury in which a water bird (cormorant species) had pecked his eye while he was fishing. A fulminant endophthalmitis developed despite treatment with intravenous, intravitreal and topical antibiotics and initial wound repair. Enucleation was performed. RESULTS: Aeromonas sobria was isolated from the vitreous aspirate. CONCLUSION: Aeromonas sobria infection should be suspected in water-contaminated penetrating eye injuries. The prognosis in this case was poor. PMID- 9395835 TI - Cast-forming Actinomyces israelii canaliculitis. AB - BACKGROUND: Primary chronic canaliculitis is an uncommon disease usually caused by Actinomyces israelii (streptothrix). Actinomyces israelii is a cast-forming Gram-positive anaerobe that is difficult to isolate and identify. We present a case that demonstrates the typical clinicopathological presentation of this unusual condition and discuss management options. METHODS AND RESULTS: A 10-year old girl presented with a 6 month history of intermittent 'conjunctivitis' and discharge from her 'pouted' left lower punctum. Microbiology confirmed probable A. israelii infection, but topical treatment failed. Exploration under anaesthesia revealed a canalicular diverticulum and three canaliculiths. Histological examination of the canaliculiths demonstrated that they consisted of solid casts of Actinomyces. Punctoplasty, removal of the casts, and adjunct antibiotic therapy resulted in resolution of the canaliculitis. CONCLUSIONS: Primary chronic canaliculitis should be considered in any patient who presents with chronic or recurrent conjunctivitis and the eyelid should be inspected for a discharging and 'pouting' punctum. Failure of the condition to resolve on topical treatment requires surgical exploration of the canalicular system and removal of any casts. Extensive surgery is not always required. PMID- 9395836 TI - The pharmacokinetics of bumetanide in the newborn infant. AB - This study characterizes the pharmacokinetics of bumetanide after an intravenous dose of 0.05 or 0.10 mg/kg to 14 neonates (weight range 820-4,000 g; gestational age 26-40 weeks) during the first week of life. Blood samples and urine were collected for up to 12 h after dosing. Estimated serum clearance was 0.2-1.0 ml/min.kg (range), volume of distribution was 0.22 l/kg (range 0.11-0.32 l/kg), and the harmonic mean half-life was 6-7 h (range of 4-19 h). Nonrenal clearance accounted for 58-97% of the serum clearance with the presence of certain oxidative metabolites of bumetanide in the urine. These findings suggest higher dosing requirements and prolonged intervals as compared to adults. Utilizing these pharmacokinetic data, pharmacodynamic and ototoxicity studies should be conducted to establish a safe and effective neonatal dose. PMID- 9395837 TI - Interplay between glutathione-S-transferase and glucose-6-phosphate dehydrogenase in neonatal cord blood. AB - Three hundred neonatal cord blood samples were collected from two major cities in Jordan and assayed for glutathione-S-transferase (GST), glutathione, and glucose 6-phosphate dehydrogenase (G6PD). A significant positive correlation between the levels of G6PD and GST activities was detected. When the G6PD activity decreased, the GST activity declined. A similar concurrent decrease in the concentration of reduced glutathione was also observed. When the samples were divided into males and female samples, some significant variations in the levels of the two enzymes were observed. Female samples exhibited higher G6PD and GST activities as compared with male samples. Moreover, the incidence of severe and moderate G6PD deficiencies was higher in male as compared with female samples. Analyses of the samples for total bilirubin, red blood cells, hematocrit, and hemoglobin were also performed, and slight nonsignificant variations were noted. PMID- 9395838 TI - Hematological features of fetal triploidy: a report of 11 cases. AB - Using fetal blood sampling, the diagnosis of triploidy can be strongly suspected on the basis of the peculiar hematological picture. Triploid fetuses present with anemia, marked anisopoikilocytosis, and grossly increased mean corpuscular volume, associated with thrombocytopenia and significant platelet anisocytosis. These findings are of considerable immediate diagnostic value. They allow physicians to immediately counsel parents about the prognosis of the fetus. In case of fetal or neonatal distress, this information could orientate decisions about obstetrical and pediatric management while waiting for the definitive diagnosis. PMID- 9395839 TI - Response of hepatic mitochondrial and peroxisomal beta-oxidation to increasing palmitate concentrations in piglets. AB - Responses of total, mitochondrial, and peroxisomal beta-oxidation to increasing [1-14C]-palmitate concentrations (0.02-1.0 mM) were measured in liver homogenates from neonatal pigs. Incubations were conducted in the absence (total beta oxidation) or presence (peroxisomal beta-oxidation) of antimycin A and rotenone; mitochondrial beta-oxidation was calculated as total minus peroxisomal oxidation. Total and mitochondrial beta-oxidations were maximized at a palmitate concentration of 0.05 mM, whereas peroxisomal beta-oxidation was maximized at 0.50 mM palmitate. Across concentrations, peroxisomal beta-oxidation contributed 40-47% of total beta-oxidation. An increased rate of CO2 production and a greater ratio of CO2 production to total mitochondrial beta-oxidation as palmitate concentration increased suggested that the limited capacity for mitochondrial beta-oxidation was attributable primarily to limited ketogenic capacity. Comparative observations in liver from adult rats showed that peroxisomal beta oxidation was maximized at 0.1 mM palmitate, but total and mitochondrial beta oxidation rates were not maximized even at 1 mM palmitate. At 1 mM palmitate, peroxisomal beta-oxidation was 20% of total beta-oxidation in adult rats and 37% in adult pigs. Therefore, the contribution of peroxisomal beta-oxidation to total beta-oxidation is highly dependent on substrate concentration and appears to be greater in adult pigs than in adult rats. The greater proportional contribution of peroxisomal beta-oxidation in piglet liver might act as a compensatory mechanism for piglets to oxidize milk fatty acids. PMID- 9395840 TI - Chronic intermittent in utero exposure to morphine: effects on respiratory control in the neonatal guinea pig. AB - This study was done to determine if chronic intermittent in utero exposure to morphine (MOR) during the last half of gestation results in hypoventilation and decreased ventilatory sensitivity to CO2 in the neonate. Pregnant guinea pigs were randomly assigned to once-daily treatment with saline and 1.5, 5.0, or 15.0 mg/kg MOR. Neonates were studied for 3 weeks. Prenatal exposure to 5.0 and 15.0 mg/kg MOR significantly increased neonatal minute ventilation and central inspiratory drive on day 7 while breathing room air or 5% CO2. The increase in minute ventilation was part of a withdrawal syndrome that included increased locomotor activity, but was not due to an increase in metabolic rate or sensitivity to CO2. We conclude that neonatal guinea pigs exposed once daily to MOR during the last half of gestation hyperventilate during the 1st week after birth. These changes are neither permanent nor followed by hypoventilation or depressed sensitivity to CO2. PMID- 9395841 TI - Combined effects of fetal beta agonist stimulation and glucocorticoids on lung function of preterm lambs. AB - We asked whether a single-dose fetal treatment strategy using betamethasone plus either a long-acting beta 2 agonist (formoterol) or betamethasone plus agents that elevate intracellular cyclic adenosine monophosphate (isobutyl methylxanthine and dibutyryl-cyclic adenosine monophosphate) would augment the effects of prenatal betamethasone on postnatal lung function. Preterm lambs were treated with 0.5 mg/kg beta-methasone or betamethasone plus the other agents and delivered 48 h after treatment. The postnatal lung function as assessed by compliance, ventilatory efficiency, and lung volumes at 40 min of age was improved by prenatal betamethasone and improved further by combination treatment, although the augmented responses were not significantly greater than with betamethasone alone. Fatty acid synthase protein and enzymatic activity were not increased by betamethasone or combined treatments, in contrast to responses reported for other animal models. There were no effects of glucocorticoids or the combined treatments on surfactant. Stimulation of the beta 2 agonist system did not augment postnatal lung function significantly above that noted for betamethasone alone with the agents, doses, and duration of exposures tested. PMID- 9395842 TI - Estrogen-regulated uterine vascularization modulates the guinea pig intrauterine environment. AB - The effects of experimentally-induced, uterine vascular restriction on uterine blood flow (UBF) and uterine blood volume (UBV) capacity, as well as the dependent intrauterine oxygen tension (IUpO2) measurements used as an indication of luminal nutrient availability, were examined using ovariectomized, estrogen (E)-treated guinea pigs. Following 3 days of E treatment, both UBF and UBV measurements were found to be elevated and associated with a causally-related increase in intraluminal uterine oxygen availability levels. Following the acute clamping of the uterine arteries, both UBF and UBV levels decreased dramatically and induced a rapid fall in associated intrauterine luminal oxygen tension measurements. As a result of chronic (i.e., 6 h) restriction of segmental blood flow to the uterus by vascular cauterization, both UBV and IUPO2 levels were suppressed as compared with sham-operated control levels, whereas UBF rates were not significantly altered. The results of the present studies are the first quantitative demonstration that either acute or chronic reductions in uterine vascular capacity or competency can induce rapid and dramatic changes in the intrauterine nutritional environment recognized to be essential for the initiation, support and maintenance of nidation and subsequent fetal-placental development. PMID- 9395843 TI - Assessment of endogenous nitric oxide formation in newborns: measurement of urinary nitrite and nitrate concentrations. AB - We measured the urinary nitrite and nitrate (NOx-) excretion, an index of endogenous nitric oxide formation, in term and preterm newborns on the 1st and 4th days of age. In the infants of both groups, the urinary NOx- excretion significantly increased from the 1st to the 4th day. The urinary NOx- excretion in preterm infants was significantly higher as compared with term babies on both days. Furthermore, the urinary NOx- excretion was significantly elevated in preterm infants with respiratory distress syndrome as compared with those without cardiopulmonary complications on the 4th day. These changes of urinary NOx- excretion in newborns strongly suggest the presence of an active physiological role for nitric oxide in the circulatory adaptation to extrauterine life. PMID- 9395844 TI - Risk factors of sensorineural hearing loss in preterm infants. Biol Neonate 1997;71:1-10. PMID- 9395845 TI - Interference between two concurrent tasks is associated with activation of overlapping fields in the cortex. AB - Interference between two concurrent tasks can be measured as an increased reaction time during simultaneous performance compared to when each task is performed alone. We tested the hypothesis that two tasks interfere because they require activation of overlapping areas of the cerebral cortex. With positron emission tomography we measured cortical activation as fields with significant increase in regional cerebral blood flow during single task performance of an auditory and a visual go/no-go task and an auditory and a visual short-term memory (STM) task. In a separate experiment we measured the degree of interference between the two go/no-go tasks and between the two STM tasks during dual task performance. Both the two go/no-go tasks and the two STM tasks activated overlapping parts of the cortex and interfered significantly during dual task performance. The two STM tasks had a larger volume of overlap and also significantly larger increase in reaction time during dual task performance, compared to the go/no-go tasks. The results thus indicate that two concurrent tasks interfere, with a resulting increase in reaction time, if they require activation of overlapping parts of the cortex. PMID- 9395846 TI - Visual evoked cortical magnetic fields to pattern reversal stimulation. AB - We studied visual evoked magnetic fields to pattern reversal stimulation in six healthy subjects. Similar to the N75-P100-N145 components in visual evoked potentials, triphasic deflections, N75m-P100m-N145m, were clearly observed around the midoccipital position. A very small component, P50m, was occasionally observed preceding the N75m. Equivalent current dipoles (ECDs) of the main deflection, P100m, to quadrant-field stimulation were estimated near or around the calcarine fissure contralateral to the stimulation. The vertical ECD location of the P100m to the upper quadrant-field stimulation was estimated significantly lower (0.81 +/- 0.45 cm) than those to lower stimulation. These results were compatible with the retinotopic organization of the visual cortex (cruciform model) and suggested that the P100m originated in the striate cortex. The small P50m, although only a small number of ECDs could be estimated reliably, was located in the contralateral visual cortex. ECDs of the N75m were estimated mainly near or around the contralateral calcarine fissure. ECDs of the N145m were estimated also retinotopically, but with a greater vertical distance (2.90 +/- 1.09 cm) between upper and lower quadrant-field stimulation. MR-overlaid ECDs of the N145m suggested that these originated in the extrastriate cortex. No ECD was estimated when a probe was placed at the midfrontal position. PMID- 9395847 TI - Operant performance and cortical acetylcholine release: role of response rate, reward density, and non-contingent stimuli. AB - The relationship between acetylcholine (ACh) efflux in medial prefrontal cortex (mPFC) and performance in a visual discrimination task and a variable interval (VI) schedule of reinforcement was studied in rats. Animals were pretrained in one of the two tasks and then unilaterally implanted with microdialysis guide cannula into the mPFC. Animals were then dialyzed, during 12 min collection intervals, in the operant chambers prior to task onset and during and after task performance. Each animal was dialyzed for a total of four sessions: two standard task sessions, one session in which a houselight was flashed at 0.5 Hz during the third 12 min block, and an extinction session (always the last session) in which reinforcement was withheld during the final three blocks. Response accuracy in the discrimination task was very high (> 95% correct) and stable across the four blocks with a progressive increase in omissions. The flashing houselight did not affect performance whereas the loss of reinforcement led to an increase in omissions. VI performance was associated with a high number of lever presses and a high reward rate that declined over the four blocks. Again, the flashing houselight did not affect VI performance whereas lever pressing declined markedly during the extinction session. ACh efflux did not change, relative to baseline, during performance in either task, or with the presentation of the flashing houselight or the loss of reinforcement. These data contrast with the changes in cortical ACh efflux observed in situations characterized by the presentation of novel stimuli or changing demands on attentional processing and, therefore, assist in the specification of hypotheses on the cognitive functions of cortical ACh. PMID- 9395848 TI - How the brain processes complex words: an event-related potential study of German verb inflections. AB - Event-related brain potentials (ERPs) were recorded as German-speaking subjects read verbs in correct and incorrect participle forms. The critical words were presented in three different versions to three different groups of subjects, as part of a simple sentence, in a word list, and embedded in a story; for each version separate ERPs were recorded. Three types of verbs were investigated, regulars, irregulars and nonce verbs. We compared correct regular and irregular participles with incorrect ones; the latter had -(e)n on verbs that actually take -t participles (* getanz-en), or -(e)t on verbs that require -(e)n (* gelad-et). For the nonce verbs, we compared participles with the unexpected -(e)n ending with the expected -t participle forms. The ERP responses were very consistent across the three versions of the experiment: (i) incorrect irregular participles (* gelad-et) elicited a left frontotemporal negativity; (ii) incorrect regulars (* getanz-en) produced no differences to the correct ones; (iii) nonce verbs were associated with an N400 component but did not show a difference between expected and unexpected endings. We will interpret these findings with respect to psycholinguistic models of morphological processing and argue that the brain processes regularly inflected words differently from irregularly inflected ones, the latter by accessing full-form entries stored in memory and the former by a computational process that decomposes complex words into stems and affixes. PMID- 9395849 TI - Encoding behavioral context in recurrent networks of the fronto-striatal system: a simulation study. AB - This research addresses the hypothesis that behavioral context is encoded in recurrent networks of the fronto-striatal system. Behavioral context influences the processing of subsequent brain events, including responses to sensory inputs, thus providing a basis for context-dependent behavior. We define context dependent behavior as the adaptive ability to produce the appropriate response to a given stimulus, dependent upon the context in which it appears. Behavioral context can change with a time-scale on the order of seconds to tens of seconds or more. This suggests a flexible mechanism that encodes context via an ensemble of neural activation that will appropriately influence the processing of subsequent sensory stimuli. We present a functional model of context encoding in recurrent connections of the fronto-striatal system with simulation results that correspond closely to empirical data. Neuronal activity in monkeys that perform a context-dependent task indicate that the prefrontal cortex and striatum participate differentially in this kind of context encoding. Likewise, simulated neurons in our model of the fronto-striatal system, which performs the context dependent task, display task-related activity remarkably similar to that found in monkey frontal cortex and striatum, supporting our hypothesis. PMID- 9395850 TI - Loss of functional hemispheric asymmetry in Alzheimer's dementia assessed with near-infrared spectroscopy. AB - In a total of 10 patients with dementia of the Alzheimer-type (DAT) and in 10 healthy controls near-infrared spectroscopy (NIRS), a new non-invasive optical method, was used to measure the changes of concentrations of oxy- (O2HB) and deoxyhemoglobin (HHB) in left and right hemispheric prefrontal brain tissue areas during performance of the Verbal Fluency Test (VFT). On a neuropsychological level, the healthy subjects performed better in the VFT than patients with DAT. Statistical analysis of the relative concentrations of O2HB and HHB measured with NIRS during performance of the VFT revealed a significant interaction of the hemispheric effects with the diagnosis. A possible interpretation of this finding is that a good performance in the VFT relies on a predominantly left hemispheric activation observed in controls, whereas a low number of correct responses is associated with a loss of this asymmetric activation in patients with DAT. Although both, patients and controls, performed better in the category version of the VFT, the metabolic effects of this task were significantly less pronounced than in the letter version. This indicates that different energy demands, according to the type of access to the memory stores, may be interpreted as the result of a less energy-demanding access to categorically stored information and adds further evidence to the view that memory departments in humans are organized according to categorical principles. PMID- 9395851 TI - Comparative study of antifungal activity of sertaconazole, terbinafine, and bifonazole against clinical isolates of Candida spp., Cryptococcus neoformans and dermatophytes. AB - The in vitro activity of sertaconazole was compared with that of terbinafine and bifonazole against 180 Candida spp., Cryptococcus neoformans yeasts and 53 dermatophytes. Minimum inhibitory concentrations (MICs) were determined by a microdilution method in Sabouraud's buffered liquid medium (pH 5.6). Sertaconazole (arithmetic mean MIC 1.24 mg/l) was statistically more active than bifonazole (MIC 6.54 mg/l) and terbinafine (MIC 12.61 mg/l) against yeasts strains, MIC values for sertaconazole being generally and specifically lower for each tested yeast species. MIC for C. parapsilosis (0.26 mg/l) demonstrated a higher activity of sertaconazole against this species, in contrast to C. tropicalis (MIC 1.49 mg/l). Against dermatophytes, MIC for terbinafine (0.05 mg/l) was lower than sertaconazole (MIC 0.41 mg/l) and bifonazole (MIC 1.04 mg/l). These results, obtained under the same experimental conditions, confirm the good antifungal activity of sertaconazole against both yeasts and dermatophytes with lower MICs obtained in the topical application. This in vitro activity correlates with the clinical efficacy of sertaconazole compared with other antifungal agents. PMID- 9395852 TI - In vitro activity of biapenem against recent gram-negative and gram-positive clinical isolates. AB - The in vitro activity of biapenem, a new carbapenem, against 535 clinical recent isolates was compared with those of other antibiotics. Biapenem showed broad spectrum activity against gram-negative and gram-positive clinical isolates. The new carbapenem was more active than imipenem against members of the family Enterobacteriaceae with MIC90S ranging from 0.12 to 2 mg/l and from 0.25 to 4 mg/l, respectively. Moreover it was 2-fold more active than imipenem against Pseudomonas aeruginosa (MIC90, 8 and 16 mg/l, respectively). Taken together, these results indicate that biapenem shares the favorable in vitro activity properties of imipenem and merits further study in the treatment of infections caused by a wide range of pathogens. PMID- 9395853 TI - Bacterial colonization of the upper respiratory tract of patients with primary lung cancer and nonmalignant lung disease. AB - We examined the bacterial colonization of the upper respiratory tract of 110 patients with primary lung cancer (PLC), 75 patients with nonmalignant lung diseases (NMLD) and 45 healthy volunteers (HV), comparing the sensitivity of expectorated sputum, and throat and nasal swabs. The frequency of bacterial colonization of the upper respiratory tract was significantly higher in the PLC patients (59.1%) than in NMLD patients (37.3%, p < 0.01) and HV (37.8%, p < 0.01). The frequency of gram-negative colonization was significantly higher in PLC patients than in the other subjects (p < 0.01). Expectorated sputum and nasal swab were the most sensitive for detection of whole bacteria and methicillin resistant Staphylococcus aureus in the patients with PLC. Our results showed that PLC patients are significantly more frequently colonized by bacteria in their upper respiratory tracts and that a combination culture of expectorated sputum and nasal swab is suitable to estimate the bacterial colonization of the upper respiratory tract in the patients. PMID- 9395855 TI - Protein kinase C inhibitors augment tumor-necrosis-factor-induced apoptosis in normal human diploid cells. AB - In this report we show augmentation of tumor necrosis factor (TNF)-induced apoptosis by protein kinase C (PKC) inhibitors in human embryonic lung fibroblast (HEL) cells. Staurosporine (STS) reportedly potentiates TNF-mediated cytotoxicity in cancer cell lines via inhibition of PKC. We investigated whether STS or another potent PKC inhibitor, 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H7), augmented TNF-induced apoptosis in normal human diploid cells and examined the effects of the PKC inhibitors on the expression of endogenous TNF (enTNF) and manganous superoxide dismutase (MnSOD) as possible cell resistance factors opposing TNF-induced apoptosis. Both PKC inhibitors augmented TNF-mediated DNA fragmentation in HEL cells, which are normally TNF resistant and constitutively express high amounts of enTNF and MnSOD activity. Neither the number nor the affinity of TNF receptors were altered by STS and H7. The expression of enTNF and MnSOD was suppressed by the presence of STS or H7 along with TNF. The results indicate that PKC inhibitors augment TNF-induced apoptosis not only in tumor cells but also in normal cells by inhibitory effects on cellular resistance factors such as enTNF and MnSOD. PMID- 9395854 TI - Endogenous tumor necrosis factor functions as a resistant factor against hyperthermic cytotoxicity in pancreatic carcinoma cells via enhancement of the heart shock element-binding activity of heart shock factor 1. AB - To elucidate the relationship between two distinct resistant factors, endogenous tumor necrosis factor (enTNF) and heat shock proteins (HSPs), against hyperthermia, we assessed whether enTNF enhances HSP72 expression. Although there was a variability in the sensitivity of pancreatic carcinoma cell lines to heat, enTNF and HSP72 expression as well as MnSOD activity correlated positively with heat resistance. When MIAPaCa-2 pancreatic carcinoma cells, which had the lowest enTNF expression and highest heat sensitivity, were transfected with a nonsecretory-human TNF gene (pTNF delta pro), intracellular manganous superoxide dismutase (MnSOD) activity, HSP72 expression, and heat resistance were significantly increased. Furthermore, in these cells, enTNF expression correlated with the binding activity of heat shock factor 1 (HSF 1) to an oligonucleotide containing the human heat shock element. These results indicate that enTNF participates in the intrinsic resistance against heat via induction of MnSOD, enhances HSF1-binding activity, and augments of HSP72 expression. Therefore, inhibition of enTNF expression in pancreatic carcinoma cells would improve the efficacy of hyperthermia for pancreatic carcinoma. PMID- 9395856 TI - Inhibition of motility and adherence of Proteus mirabilis to uroepithelial cells by subinhibitory concentrations of amikacin. AB - The effect of subinhibitory concentrations of amikacin on Proteus mirabilis motility and adherence to human uroepithelial and to HeLa cells was compared with that of gentamicin. In addition, the effect of both antibiotics on cell surface hydrophobicity was also examined. Exposure of bacterial cells in the logarithmic phase to one fourth of amikacin or gentamicin at one fourth of their respective minimal inhibitory concentrations causes the inhibition of swarming and motility of Proteus strains. Amikacin significantly reduced adhesion of Proteus strains to human uroepithelial cells and gentamicin exerts the same effect to a lesser extent. Such inhibitory concentrations of amikacin or gentamicin had no significant effect on the attachment ability of these strains to HeLa cells compared to the nontreated cells. Treatment of the bacterial cells with amikacin or gentamicin changed significantly the cell surface hydrophobicity towards the hydrophilic state compared to nontreated cells, and it was found to be strain dependent. Since motility and attachment ability are considered as pathogenic traits, these data indicate the impact of amikacin on the virulence factors especially in urinary tract infections with Proteus strains. PMID- 9395857 TI - Improvement of albendazole efficacy against enteral, but not against parenteral stages of Trichinella spiralis by preparing solid dispersions in polyvinylpyrrolidone. AB - A comparison was made, in the Trichinella/mouse model, of the anthelmintic effects of albendazole (ABZ) and ricobendazole (RBZ) formulated as solid dispersions in polyvinylpyrrolidone with regard to ABZ formulated as a suspension in carboxymethylcellulose. A solid dispersion significantly increased (p < 0.01) the efficacy of the drugs against intestinal preadult but not against migrating and muscle stages of the parasite. The anthelmintic efficacy of RBZ given as a solid dispersion was equivalent to (against preadult and encysted larvae) or significantly lower than (against migrating larvae) that of ABZ with the same formulation. The pharmacokinetic profiles of ABZSO as measured by HPLC showed no significant differences in the Cmax and AUC following administration of ABZ formulated as a suspension or solid dispersion although the Tmax was significantly lower for the dispersion. PMID- 9395858 TI - Combination therapy with fluconazole and flucytosine for pulmonary cryptococcosis. AB - We investigated, in vitro, the combined effects of fluconazole (FLCZ) and flucytosine (5-FC) against different strains of Cryptococcus neoformans, and retrospectively analyzed the clinical efficacy of combination therapy of FLCZ and 5-FC in patients with pulmonary cryptococcosis. The minimum inhibitory concentrations (MICs) of antifungal agents and drug interaction were determined by the broth microdilution method and checkerboard titration. FLCZ and 5-FC showed synergistic activity against 8 (32%) of 25 strains of C. neoformans. The clinical efficacy of the 2 drugs when combined together was good in 9 (90%) patients and fair in 1 (10%) patient with pulmonary cryptococcosis. Renal dysfunction occurred in 1 patient. Our results suggest that a combination therapy using FLCZ and 5-FC is clinically useful in patients with pulmonary cryptococcosis who otherwise show a limited response to monotherapy. PMID- 9395859 TI - Experience with once daily dosing of gentamicin: considerations regarding dosing and monitoring. AB - Plasma concentrations of gentamicin following a fixed dose of 240 mg once daily to patients with normal renal function were measured. The purpose was to establish guidelines to achieve a sufficiently high peak concentration with an appropriately low risk of accumulation. In 40 patients, 1-hour concentrations of plasma gentamicin had a median of 9.3 mg/l (range: 4.5-19.0 mg/l) and 9.7 mg/l (range: 3.6-14.6 mg/l) on days 1 and 3 of gentamicin treatment, respectively. Thirty-nine patients had 1-hour concentrations > 5 mg/l. The 1-hour concentrations varied considerably intra- and interindividually but showed a significant inverse correlation with body weight, surface area and the estimated endogenous creatinine clearance. The plasma gentamicin elimination half-life correlated significantly with age and inversely with body weight and creatinine clearance. There was no increase in the mean plasma creatinine from day 0 to day 4. No patients showed signs of nephrotoxicity, although 2 patients, both elderly and with low body weight, showed signs of beginning gentamicin accumulation. In conclusion, gentamicin treatment with the dose of 240 mg once daily in 3 days to adults with normal kidney function generally does not require adjustment or monitoring. However, the dose should be increased in young patients with an excessive body weight, and decreased doses are needed for old and underweight patients. Monitoring of trough plasma gentamicin concentration is not necessary with treatment duration of 3 days or less. PMID- 9395860 TI - Improbability of selection for RIF resistance in Mycobacterium tuberculosis by accidental exposure during short-course therapy with Cotrifazid. PMID- 9395862 TI - Catastrophic metabolic encephalopathies in the newborn period. Evaluation and management. AB - The newborn who presents with neurologic symptoms such as seizures or lethargy due to inborn error of metabolism is an important problem. Although each inborn error that presents in this manner is rare, these conditions are not rare as a group, and more than one in 1000 babies is affected with one of the more than 100 different inborn errors that are now known. Many of these conditions present with much the same features seen in sepsis or asphyxia and, when untreated, can lead rapidly to death or permanent neurologic damage. Early diagnosis and management may prevent some or all of this morbidity, and also permits the parents to be informed about the chances of having other affected children. Despite the large number and complexity, most metabolic encephalopathies can be diagnosed by applying a few simple clinical principles and laboratory tests. These principles, and the typical features of some inborn errors that present in the neonate, are detailed in this article. PMID- 9395861 TI - Seizures in the newborn infant. Diagnosis, treatment, and outcome. AB - Neonatal seizures remain an emergent clinical problem in the neonatal intensive care unit that requires prompt diagnosis and treatment. The electroencephalogram is the preferred tool by which a surface-recorded seizure can be documented. While a number of etiologic possibilities may occur, overlapping mechanisms result in a lower seizure threshold. The susceptibility to seizures shortly after birth, efficacy of antiepileptic medications to control seizures, as well as the prediction of outcome in patients with neonatal seizures remains controversial. Prospective studies are required to assess interactions among maternal, fetal, and neonatal conditions that contribute to the occurrence of neonatal seizures. PMID- 9395863 TI - Infections of the central nervous system in the newborn. AB - Safe, effective vaccines and potent antimicrobial agents have diminished substantially the morbidity and mortality associated with neonatal infections of the central nervous system (CNS), and new molecular methods, such as the polymerase chain reaction, enable clinicians to detect micro-organisms rapidly. Despite these advances, CNS infections remain an important cause of death and neurodevelopmental sequelae. This article summarizes current concepts regarding infections of the developing CNS. PMID- 9395864 TI - Neurologic complications of cardiac disease in the newborn. AB - Advances in the management of infants with congenital heart disease have lead to a striking decrease in mortality. The most dramatic impact has been in the newborn infant with complex and previously lethal heart disease. These heart lesions have become amenable to corrective procedures in the newborn period because of the development of support techniques such as low-flow cardiopulmonary bypass and deep hypothermic circulatory arrest. However, these techniques have demonstrated their own inherit risk for neurologic injury. Consequently, in recent years there has emerged a growing population of infants surviving congenital heart disease only to manifest subsequent neurologic complications originating from injury in the hemodynamically unstable preoperative period or periods of intraoperative hypoperfusion. The preservation of neurology function has emerged as the next frontier in the management of congenital heart disease. Finally, neurologic dysfunction in the newborn with cardiac disease may reflect associated brain malformations or the combined cardiac and brain manifestations of inherited metabolic disease. The clinical features and mechanisms of brain injury are discussed for these structural and metabolic cardiac diseases presenting in the newborn infant. PMID- 9395865 TI - Neuromuscular disorders in the newborn. AB - The main manifestations of neuromuscular disease in the newborn period are hypotonia and weakness. Infants with severe hypotonia but only marginal weakness usually do not have a disorder of the lower motor unit. These infants may have genetic conditions, metabolic disturbances, congenital heart disease, hypothyroidism, sepsis, or other systemic disorders. Early on, neonates with central nervous system pathology may present with profound hypotonia, decreased reflexes, and moderate to severe but transient weakness. However, they also tend to have seizures, obtundation, cranial nerve signs, or history of perinatal asphyxia. PMID- 9395866 TI - Neurologic birth trauma. Intracranial, spinal cord, and brachial plexus injury. AB - The variety of perinatal neurologic injuries described in this article suggests that any region of the central or peripheral nervous system may be affected by the birth process. Fortunately, these injuries are infrequent and, in many instances, resolve without any intervention. PMID- 9395867 TI - Brain death in the newborn. Current perspectives. AB - Brain death can be diagnosed in the full-term newborn, even when less than 7 days of age. An observation period of 48 hours is recommended to confirm the diagnosis. If an EEG is isoelectric or if a CBF study shows no flow, the observation period can be shortened to 24 hours. Although there are few cases of preterm infants who are brain dead, it is likely that the same time frame would be applicable. Based on available data, the risk of misdiagnosis appears exceedingly low. There have been few instances of neonates or older infants who showed minimal transient clinical or EEG recovery but with no meaningful neurologic function and all died within brief periods of time. PMID- 9395868 TI - The Castroviejo Lecture. Prolonged eyelid closure is a risk to the cornea. AB - PURPOSE: The author introduces new concepts and summarizes published evidence suggesting that prolonged eyelid closure puts the cornea at risk. METHODS: Significant clinical and experimental publications are reviewed, and the author's experience is applied relating to the pathogenesis and treatment of a variety of clinical entities thought to be induced, to some degree, by prolonged eyelid closure. RESULTS: Evidence from the scientific literature suggests that the hypoxia or reduced tear volume or both that result after prolonged eyelid closure, especially during sleep and when a soft contact lens is in place, serve as risk factors in the "sucked-on" contact lens syndrome, recurrent corneal erosion, chronic corneal deepithelialization, Pseudomonas keratitis, filamentary keratitis, superior limbic keratoconjunctivitis, sterile midperipheral corneal infiltrates, and corneal vascularization. The limbal stem cells under the upper eyelid are subjected to continuous hypoxic stress and are at special risk to other insults. CONCLUSIONS: Prolonged eyelid closure, such as in patching and in sleep, is a risk factor in the pathogenesis of a variety of corneal conditions. PMID- 9395869 TI - A study of 530 patients referred for rigid gas permeable scleral contact lens assessment. AB - PURPOSE: The purpose of this study was to analyse the current application of scleral contact lenses in two specialist centres. METHODS: The case notes of 530 patients assessed for fitting or refitting with rigid gas permeable (RGP) scleral lenses were retrospectively analysed to determine the indication for contact lenses and the outcome. Scleral lenses had been offered as a conservative management option in suitable cases for a variety of visual and medical indications. RESULTS: Various types of primary corneal ectasia, ranging from low grade to advanced, including keratoconus, keratoglobus, and pellucid marginal degeneration, formed 53.0% of the total referred for assessment. The other principal indications for contact lenses were corneal transplant (15.8%), aphakia (10.3%), high myopia (8.9%), and various ocular surface disorders (8.2%). Sixty percent continued to use scleral lenses, 42.9% RGP, and 17.1% polymethylmethacrylate lenses. Twenty-two percent discontinued scleral lens wear or failed a trial of scleral lenses, with 9.3% in progress at the time of assessment and 8.7% lost to follow-up. CONCLUSIONS: In the authors' opinion, scleral lenses have retained their traditional role in the management of complex ametropia and ocular surface disease. That role has been further enhanced by the application of gas permeable materials. PMID- 9395870 TI - Indications for penetrating keratoplasty and associated procedures, 1989-1995. AB - PURPOSE: To identify changing trends in indications for penetrating keratoplasty and associated surgical procedures. METHODS: Review of charts from all patients who underwent penetrating keratoplasty at Wills Eye Hospital from January 1, 1989 through December 31, 1995. RESULTS: A total of 2,442 corneal transplants were performed in 2,186 patients. The leading indication for penetrating keratoplasty was pseudophakic corneal edema, accounting for 634 cases (26.0%); 54.7% of them were associated with anterior chamber intraocular lenses, 36.4% with posterior chamber intraocular lenses, and 3.1% with iris-fixated intraocular lenses. Regraft (17.8%), Fuchs' dystrophy (15.7%), and keratoconus (13.2%) followed pseudophakic corneal edema in frequency. Cataract extraction, with or without intraocular lens implantation, was combined with penetrating keratoplasty in 439 cases of 1,264 phakic eyes (34.7%). Intraocular lens exchange was performed in 285 of the 634 cases of pseudophakic corneal edema (44.9%). CONCLUSION: Pseudophakic corneal edema was the leading indication for penetrating keratoplasty, with an increasing number of cases associated with posterior chamber intraocular lenses during the study period (p = 0.001). The number of regrafts steadily increased between 1989 and 1995 (p = 0.001), being the second most common indication for corneal transplantation since 1992. PMID- 9395871 TI - Effects of artificial tear temperature on corneal sensation and subjective comfort. AB - PURPOSE: Cooling reduces acute inflammation and local nerve sensation. We investigated the relationship between artificial tear temperature, ocular surface sensation, and patient comfort. METHODS: We placed preservative-free artificial tears and eye mask stored at four temperatures (36 degrees C, 25.2 degrees C, 4 degrees C, and -10 degrees C) in the right eyes of 24 normal subjects, whose left eyes served as controls. Corneal and conjunctival sensations were measured and corneal temperature was recorded. Comfort was reported on a 7-point scale. RESULTS: Corneal temperature was significantly lowered with all temperature artificial tears and frozen eye mask (p < 0.001 for each temperature relative to the previous one). Aesthesiometer readings were inversely correlated with corneal temperature (r = -0.45, p = 0.0005), decreasing with lower temperatures, reaching 2.0 +/- 1.3 g/mm2 (p = 0.001) for the mask. Conjunctival sensation reacted similarly and was well correlated with both corneal temperature (r = 0.43, p = 0.0009) and corneal sensation (r = 0.39, p = 0.006). Treatments provided relief, with the 4 degrees C tears being the most comfortable (p = 0.0001). CONCLUSION: Although there may still be some biases, cooled artificial tears provide relief to the eye by the mechanism of reduced corneal and conjunctival sensation. PMID- 9395872 TI - Granular-lattice (Avellino) corneal dystrophy in Japanese patients. AB - PURPOSE: To determine amyloid deposition in the corneas of granular corneal dystrophy in Japanese patients. METHODS: Eight Japanese patients (10 eyes) with a clinical diagnosis of granular corneal dystrophy were investigated clinically and histologically. Each specimen obtained at surgery was stained with hematoxylin eosin, Masson trichrome or Mallory, and Congo red stain. Amyloid deposit was identified by birefringence and dichroism under cross-polarized light after staining with Congo red. RESULTS: Seven (70%) of the 10 corneal buttons (six of eight patients) had amyloid deposits, as shown by Congo red staining with birefringence and dichroism. Of the six amyloid-positive patients, two patients (who were siblings) showed discrete gray-white corneal deposits with additional linear deposits. This finding is typical of Avellino corneal dystrophy. The corneas of the remaining four patients showed the discrete deposits typical of granular dystrophy. Some of them showed a few whitish fusiform and stellate opacities in the mid stroma, suggestive of Avellino corneal dystrophy. CONCLUSION: The high frequency of amyloid deposits in Japanese patients with granular corneal dystrophy may be caused by an allelic heterogeneity of the gene. PMID- 9395873 TI - Evaluation of central and peripheral corneal thickness with ultrasound biomicroscopy in normal and keratoconic eyes. AB - PURPOSE: Our study was designed to calculate central and peripheral corneal thickness in patients affected with early stages of keratoconus and in normal subjects using ultrasound biomicroscopy (UBM). To obtain an objective and reliable assessment of the corneal thinning in affected eyes, we developed a keratoconus index (KI) by means of the UBM measurements. METHODS: By means of the commercial version of the ultrasound biomicroscope (system model 840; Zeiss Humphrey Instruments, San Leandro, CA, USA) using a 50-MHz probe, we studied 30 normal and affected eyes. In keratoconic eyes, we measured the thinnest corneal thickness (TCT) at the apex of the conus and at four peripheral sites at a distance of 2.5 mm from the central site (peripheral corneal thickness: PCT). The same procedure was performed in the normal eyes. To obtain an objective and reliable assessment of the corneal thinning, we calculated the ratio between the mean PCT and the mean TCT (Keratoconus Index: KI = PCT/TCT), in keratoconic eyes. In normal eyes, the KI was calculated on the basis of the ratio between the mean PCT and the mean central corneal thickness (CCT). RESULTS: In keratoconic eyes, the mean corneal thickness at the thinnest part of the conus was significantly different from the CCT in normal patients (Student's t test, p < 0.001). The peripheral measurements were not significantly different from keratoconic and normal eyes. The mean KI was 1.482 (SD, 0.095) in the keratoconic eyes, whereas it was 1.189 (SD, 0.086) in the normal subjects. The statistical analysis of the KI calculated on the basis of the UBM measurements showed that the KI values are significantly different from healthy subjects and from keratoconic patients (Student's t test, p < 0.001). CONCLUSIONS: UBM can be considered a useful tool in the study of keratoconus. We believe that calculation of the KI by means of UBM gives the possibility of obtaining an objective assessment of corneal thinning. Therefore this parameter can be useful in the staging and in the follow up of these patients. PMID- 9395874 TI - Nasolacrimal stimulation of aqueous tear production. AB - PURPOSE: Aqueous tear production decreases after anesthetizing the ocular surface. Loss of the nasolacrimal reflex is a risk factor for neurotrophic keratopathy and keratoconjunctivitis sicca. The purpose of this study was to evaluate the effect of nasal mucosal anesthesia on aqueous tear production. METHODS: Eleven healthy human volunteers with a normal ocular surface and Schirmer I tear-test scores > 10 mm participated in this study. Schirmer I values were obtained daily for 3 days to establish a normal baseline. On a separate day, the right nasal mucosa was anesthetized with aerosolized 10% lidocaine (Xylocaine). After a 10-min period to allow the anesthetic to take effect and reflex tearing to subside, the Schirmer I test was repeated. A saline nasal spray was used as a control. RESULTS: Baseline Schirmer I values for both eyes had a mean of 22.98 +/- 1.05 mm (SEM). There was no difference in Schirmer scores between the two eyes after nasal anesthesia (p > 0.6); however, when these were compared with the baseline Schirmer I values, a significant decrease in tear production was noted (p < 0.001). The mean Schirmer I value after nasal anesthesia was 15.18 +/- 1.38 mm (SEM), a 34% decrease from baseline. The difference between the baseline and the normal saline control values was not significant (p = 0.160). There was a significant difference in Schirmer test scores between the saline control and nasal anesthesia groups (p < 0.02). CONCLUSIONS: In addition to sensory neural stimulation from the ocular surface, sensory stimulation of the nasal mucosa also promotes aqueous tear production. These results may help explain the decreased tear production observed in patients who have nasal mucosal damage, disease, or denervation. PMID- 9395875 TI - Relationship between tear-meniscus parameters and tear-film breakup. AB - PURPOSE: Several flaws exist with the lipid-diffusion model for tear-film breakup. The aim of this study was to test an alternative model of tear-film rupture in which the negative hydrostatic pressure in each tear meniscus (related to the tear-meniscus radius of curvature) is proposed to influence the formation of breaks in the tear film. METHODS: Measurements of noninvasive breakup time (NIBUT) and tear-meniscus radius of curvature, height, width, and cross-sectional area (TMC, TMH, TMW, and XSA) were made for 15 aqueous-deficient dry-eye and 15 age-matched control subjects. An optic section of the inferior tear meniscus (colored with a minute volume of fluorescein) was photographed at x120 magnification, and images were computer analyzed. RESULTS: A significant positive correlation was found between log NIBUT and TMC (r2 = 0.141; p < 0.05). Furthermore, all subjects with TMC < 0.340 mm had NIBUT < 15 s, and two thirds of subjects with TMC > 0.340 mm had NIBUT > 15 s. There was a moderate linear relationship between TMH and log NIBUT, indicating an association between tear volume and tear stability. TMC, TMH, and tear meniscus XSA measurements all showed good reliability. CONCLUSIONS: The association between highly curved tear menisci and rapid tear-film breakup times is consistent with the meniscus model of tear-film rupture. However, a causal relationship has yet to be established. PMID- 9395876 TI - The antibacterial activity of topical anesthetics. AB - PURPOSE: Topical anesthetics are commonly used prior to obtaining bacterial cultures in ulcerative keratitis. We performed an in vitro study designed to test both the bacteriostatic and bactericidal effects of commercially available preserved topical anesthetic agents. METHODS: Proparacaine, tetracaine, cocaine, and sterile water solutions were applied to filter paper disks, which were then placed on Mueller-Hinton agar plates that had previously been inoculated with known quantities of Pseudomonas aeruginosa and Staphylococcus aureus. After 24 h of incubation, zones of inhibition were measured and recorded. RESULTS: Proparacaine strongly inhibited the growth of S. aureus at all concentrations (0.5%, 0.25%, 0.125%) and inhibited growth of P. aeruginosa at 0.5% and 0.25% but not at 0.125% concentration. Tetracaine also inhibited S. aureus at 0.5% and inhibited P. aeruginosa at 0.5% and 0.25% concentrations. Cocaine exhibited no inhibition of S. aureus and exhibited mild inhibition of P. aeruginosa growth only at the 4% concentration. CONCLUSIONS: The in vitro antibacterial effect of topical anesthetics suggests one possible reason why bacterial culture yields in clinical ulcerative keratitis are suboptimal. We propose that clinicians consider the use of a 1% or 2% cocaine solution instead of standard commercial topical anesthetics in the management of individual cases of ulcerative keratitis and in future clinical bacterial keratitis studies. PMID- 9395877 TI - Assessment of cornea viability by confocal laser scanning microscopy and MTT assay. AB - PURPOSE: Determination of excised cornea viability is of interest for transplant storage evaluation, but also for in vitro diffusion-study design and ocular toxicity assessment. By using simultaneous vital staining by calcein AM (CAM) and ethidium homodimer-1 (EH-1), as "live" and "dead" probes, respectively, we developed a confocal laser scanning microscopy (CLSM) assay to determine epithelial and endothelial viability and estimate cornea thickness. METHODS: New Zealand White rabbit corneas were stored in phosphate-buffered saline (PBS) or Optisol at 4 degrees C or at room temperature. At various times, corneas were stained with an EH-1/CAM solution and observed, without further treatment, by CLSM. Storage effects on the cornea were also assessed by using an MTT assay. RESULTS: Stromal swelling, shedding of the upper epithelial layers, and severe endothelial damage were observed after 4 h in PBS at room temperature. After 8 h, lower epithelial cell death was observed, along with loss of endothelial structure. Corneas stored in similar conditions in Optisol were indistinguishable from controls. Storage in Optisol at 4 degrees C affected the superficial layers of the corneal epithelium similarly at both 7 and 14 days. Extensive epithelial shedding and wing-cell death were observed at 25 days, but the basal layer remained approximately 50% healthy. Significant endothelial cell loss was observed at 25 days. MTT results were consistent with CLSM data in the medium term storage study only. CONCLUSIONS: This CAM/EH-1 CLSM fluorescence assay is a sensitive index of viability in cornea, and thus may prove useful in investigations in which maintenance of vital functions in different cell layers is critical. PMID- 9395878 TI - Alterations of corneal extracellular matrix after multiple refractive procedures: a clinical and immunohistochemical study. AB - PURPOSE: To describe the clinical course and alterations of the corneal extracellular matrix (ECM) and basement membrane (BM) in a cornea after hexagonal keratotomy, transverse keratotomies, and keratomileusis. METHODS: Frozen sections of this cornea and of 12 normal corneas were studied by immunofluorescence with specific antibodies. The patient history was analyzed to allow a clinical correlation. RESULTS: In the treated cornea, keratotomy scars and subepithelial fibrosis with neovascularization were seen. Around and beneath the epithelial plugs and along the keratotomy scars, deposits of types III, VI, VIII, and XIV collagen; fibrillin-1; fibronectin; and tenascin-C were found, together with short streaks of types IV (alpha 1-alpha 2) and VII collagen, laminin-1 and -5, entactin, and perlecan. alpha 3-alpha 4 Type IV collagen chains were abnormally absent from the BM around the epithelial plugs. At the edges of the keratomileusis flap, subepithelial fibrosis areas were found, with abnormal deposits of eight different collagen types, perlecan, fibronectin, fibrillin-1, and tenascin-C. The major part of the flap interface did not show ECM abnormalities. ECM alterations outside the scarred areas included the appearance of tenascin-C in the stroma and of alpha 1-alpha 2 type IV collagen in the epithelial BM, and the disappearance of fibronectin from Descemet's membrane. CONCLUSION: Five years after surgery, the treated cornea still presented BM abnormalities at sites of keratotomy scars and epithelial plugs. Several ECM components were abnormally expressed outside the scarred areas, consistent with an ongoing fibrosis in the treated cornea. PMID- 9395879 TI - Screening donor corneas that have undergone PRK. PMID- 9395880 TI - Posterior corneal protrusion after PRK. PMID- 9395881 TI - Corneal topography of human cadaver eyes donated months after radial and astigmatic keratotomy. PMID- 9395882 TI - Superior keratoconus. PMID- 9395883 TI - Delayed development of amiodarone keratopathy in a corneal graft. PMID- 9395884 TI - Beauveria bassiana keratitis cured by deep lamellar dissection. PMID- 9395886 TI - Analysis of DNA from subjects found to be heterozygous for the haptoglobin Johnson alpha gene. AB - We found two rare Hp Johnson variant families (Hp2-Johnson type and Hp1-Johnson type) by PAGE. It was confirmed by SDS-PAGE that the haptoglobin molecules with the Hp Johnson variant consisted of alpha-2 or alpha-1 and alpha-3 subunits. Direct gene analysis of Hp Johnson heterozygous individuals using three endonucleases: EcoRI, HindIII and BamHI, showed a three-fold tandem repeat of the same 1.7-kb DNA fragment implicated in the Hp alpha-2 gene duplication, and this Hp-Johnson-specific extension was identified in two generations of the Hp2 Johnson and Hp1-Johnson families. PMID- 9395885 TI - Fluconazole in filamentous fungal keratitis. PMID- 9395887 TI - Tumor necrosis factor-alpha gene polymorphism in psoriasis. AB - Psoriasis, an inflammatory autoimmune disease, is characterized by increased level and activity of the proinflammatory cytokine TNF-alpha in affected lesions. Two promoter region polymorphisms of the TNF-alpha locus--one at position -308 and the other at -238--were examined in 99 Caucasian patients (64 with type I and 35 with type II psoriasis) and in 123 controls. A highly significant difference in the distribution of the -238 polymorphism--the TNF (G,A) genotypes--was detected between the type I psoriasis patients and controls: compared to the controls, the frequency of the homozygous TNF-G genotype was decreased (55 vs. 91%; p = 0.0000000274; corrected p = 0.0000001644; odds ratio = 0.12), whereas that of TNF-G,A heterozygotes was increased (41 vs. 8%; p = 0.000000264; corrected p = 0.000001584; odds ratio = 7.73) in patients. No significant differences were observed in the distribution of the TNF-A homozygotes. These results suggest that homozygosity of the G allele is associated with a lower relative risk (resistance), whereas heterozygosity at this locus is associated with an increased risk (susceptibility) of type I psoriasis. PMID- 9395888 TI - Isolation of scFv fragments to polymorphic and monomorphic HLA-DR epitopes from a synthetic phage library. AB - The possibility of producing human recombinant antibodies by the phage display libraries technology has opened up new perspectives in the fields of immunological research and therapeutic applications. Despite the interest for a potential use of this technology in the HLA field, no information is so far available about selection and screening of libraries with HLA antigens. In this study we report our first experience with expression and characterization of human single-chain antibody fragments (scFvs) against HLA-DR epitopes selected from a synthetic phage library. PMID- 9395889 TI - Studies of the 6.7 family of dispersed genomic fragments within the MHC class I Region. AB - Searches for MHC-encoded disease susceptibility genes have led to considerable knowledge of the content of the class I region. In an effort to further understand the nature of the five 6.7 family members previously mapped to this region of the genome, we have further analyzed the cross-reactive members of the family and have observed additional genomic instability within the HLA-A subregion. Such genomic variation may underscore the slower evolutionary rates of the HLA-A allelic family and the extended linkage disequilibrium of markers distal to this locus. Moreover, one of the largest genes associated with a member of the 6.7 family, the 3.8-1 gene found proximal to HLA-B, was found to demonstrate limited, composite similarity to RAG2 and complement C4a gene sequences. A pancreas-specific transcript embedded in a 6.7 cross-reactive fragment was found distal to HLA-H and suggests that the fragments have remained linked to transcriptionally active chromatin comprised of both a major class I gene and a second novel coding sequence since the time of their dispersal. The absence of a 6.7 fragment in the HLA-B subregions of higher nonhuman primates lends credence to the possibility that the great apes have suffered a recent deletion event within this region following the emergence of Homo sapiens. PMID- 9395890 TI - Antibody to human MHC class I inhibits SIVsmmPBj1.9-induced proliferation of pigtailed macaque lymphocytes. AB - Previously we have shown that the simian immunodeficiency virus SIVsmmPBj1.9, a molecular clone of SIVsmmPBj14, induces proliferation of human peripheral blood mononuclear cells (PBMC). We have extended this observation to show that SIVsmmPBj1.9 induces proliferation of PBMC from pigtailed macaques. This proliferative response was markedly inhibited by mAbs against human class I MHC, class II MHC and CD4 antigens, and partially inhibited by mAbs against integrin beta 2 subunit (CD18) and LFA-1 (CD11a). However, these antibodies differed in their ability to inhibit in vitro viral infectivity of PBMC. While anti-CD4, MHC class II, and LFA-1 strongly inhibited viral infectivity, antibodies to MHC class I demonstrated little effect on viral infectivity. A control antibody (PLM2) against porcine CD18 inhibited neither virus-induced proliferation nor viral infectivity. Based on these results, we suggest that SIVsmmPBj1.9-induced proliferation requires the participation of class I MHC, class II MHC and CD4 molecules. In addition, the observation that anti-class I MHC Ab inhibited proliferation of macaque PBMC induced by mitogen (PHA) and bacterial superantigens, such as Staphylococcus enterotoxin A and toxin shock syndrome toxin-1, suggests that SIVsmmPBj1.9 also contains a viral superantigen similar to that previously demonstrated in SIVsmmPBj14. PMID- 9395891 TI - HLA class I and II genes in relation to the genetic structure and epidemiology of an Italian province. AB - The genetic structure of Pavia province is studied separately with HLA class I and II alleles. The analysis reveals that HLA class I genes reflect the same geographic barriers to migration found also with surnames, while class II gene distribution does not seem to be influenced by the presence of these barriers. The independence of HLA class II allelic distribution from the genetic structure of the area reinforces the hypothesis that environmental factors, rather than genetic drift, influence the frequency of these alleles. An example may be the significant correlation found between the distribution of the DR3-DQ2 haplotype and the prevalence of celiac disease in the province's districts. PMID- 9395892 TI - Tumor necrosis factor-alpha, interleukin-1-beta and immunoglobulin (GM and KM) polymorphisms in leprosy. A linkage study. AB - In order to determine the genetic components of susceptibility to leprosy in 6 multiplex French Polynesian families, linkage analysis was carried out between a putative disease gene and 6 polymorphic loci: G1M, G2M, KM, IL-1 beta, TNF-alpha (1, 2) and TNF-alpha (A, G) using the lod score method. The three modes of inheritance, assuming a full penetrance value or reduced penetrance values (80 and 40%) for the susceptible allele, as well as with affected ones only, were tested. The results of this study provide no evidence for linkage between leprosy and the markers tested. PMID- 9395893 TI - Nucleotide sequence comparison of the IgE constant region in patients with atopic dermatitis and non-atopic individuals. AB - In order to answer the question whether the Fc portion of the IgE molecule in patients with atopic dermatitis is altered by somatic replacement mutations, we amplified and sequenced the respective C epsilon 2 and C epsilon 3 domain genes. Five patients with atopic dermatitis and 6 non-atopic individuals were studied. Neither within the C epsilon 2 nor the C epsilon 3 domain could any common nucleotide substitutions be detected. Therefore, the conclusion can be drawn that, in patients with atopic dermatitis, there are no protein sequence differences within the Fc part of the IgE which could be responsible for distinct functional features with regard to Fc epsilon-receptor binding and signal transduction or could account for the frequent occurrence of anti-IgE autoantibodies in these individuals. PMID- 9395894 TI - The SCID mouse mutant: definition and potential use as a model for immune and hematological disorders. AB - Mice homozygous for a SCID mutation (SCID mice) are severely deficient in T and B lymphocytes. The absence of effector T and B cells has encouraged investigators to attempt engraftment of SCID mice with human fetal tissues, mature lymphocytes, hematopoietic progenitors and tumors. SCID mice can be reconstituted with human lymphocytes and are of interest for studying normal and abnormal lymphocyte development and function. SCID mice are also providing an in vivo model of infectious diseases. In addition, SCID mice readily support normal and pathologic human hematopoiesis differentiation and is useful for testing innovative hematological disease therapy. SCID mice with a fully functional human immune or hematopoietic system therefore seem to be extremely valuable for biomedical research. PMID- 9395895 TI - Sequential intravenous-oral ciprofloxacin plus amoxycillin/clavulanic acid shortens hospital stay in infected non severe neutropenic patients. AB - The objective of this study was to determine the efficacy and safety of the combination ciprofloxacin plus amoxicillin/clavulanic acid as an empirical treatment of infection in hematologic patients without severe neutropenia. These drugs allowed us to carry out a sequential therapy, first intravenously and later orally, so that the patient could be discharged as soon as there was a response. Serum concentrations of ciprofloxacin were monitored in this study. Forty seven of the sixty-six patients included (71%) responded to the treatment with no differences between the dosages of ciprofloxacin employed (600-900 mg daily in two or three divided doses). In the patients who responded, the signs and symptoms of infection lasted only three days, which could allow a short hospital stay (median of six days). In the first pre and post-dose serum samples, ciprofloxacin concentrations were significantly higher when the drug was administered every 8 h. Nevertheless, 72 h after the beginning of treatment, they had leveled out in either 8 and 12 h schemes. The toxicity of the treatment was very light, with only four cases with adverse effects, grades I and II. This data suggests that the employed combination is effective and safe and can considerably decrease costs incurred through the admission of hematologic patients with serious infections but without severe neutropenia. PMID- 9395896 TI - Fibrinolytic response to venous occlusion in patients with homozygous sickle cell disease. AB - The fibrinolytic potential was evaluated in 37 patients with homozygous sickle cell disease and compared to a control group of 30 age- and sex-matched healthy volunteers. In all individuals, the euglobulin clot lysis time and plasma antigen levels of t-Pa and PAI-1 were measured before and after venous occlusion (v.o) for 10 min. The global fibrinolytic activity was normal in 4 patients (good responders to v.o), while it was decreased in 33 patients (poor responders to v.o). Among the latter, 22 patients had significantly increased baseline levels of PAI-1 Ag (82.6 +/- 27.5 ng/ml, p < 0.001) and a normal release of t-Pa Ag after v.o. In contrast, 11 patients had basal values of PAI-1 Ag comparable to those in controls with a defective release of t-Pa Ag after v.o (11.4 +/- 5.2 ng/ml, p < 0.01). These data provide evidence for reduced fibrinolytic capacity resulting from either increased basal levels of PAI-1 or defective release of t PA. PMID- 9395898 TI - In vitro production of human antigen presenting cells issued from bone marrow of patients with cancer. AB - In the prospect of producing autologous antigen presenting cells (APC) to actively immunize patients with cancer against their own tumor we were interested in the in vitro generation of MC and/or dendritic cells. We observed that the best yielding in CD14+ cells was obtained by adding SCF and GM-CSF into RPMI 1640 completed medium and by using Teflon bags as culture-containers. The others growth factors tested (LIF, IL3 and M-CSF) were useless in term of production of macrophages. After a month of culture we usually obtained an average of 80% of CD14, CD33, CD64, CD11a, CD11b, CD11c and HLA-DR positive cells expressing the MGG staining phenotype of MC. For DC the best association of growth factors combined GM-CSF, IL-4 and SCF. Hence we could obtained at least 60% of CD1a+, CD14-, CD54+, CD58+, CD80+ and HLA DR+ dendritic cells. PMID- 9395897 TI - Red cell and platelet kinetics in chronic cytopenias following transplantation. AB - Thirty seven patients with unexplained anemia and/or thrombocytopenia after bone marrow, kidney, liver or heart transplantation were referred to the Department of Nuclear Medicine for erythrocyte or platelet kinetic studies in order to determine the mechanism of the cytopenia: accelerated destruction, or production defect. We observed only one definite case of thrombocytopenia due to accelerated autologous platelet destruction, while the life span was normal in the other 16 cases. Anemia was due to accelerated hemolysis in 7 cases, while the red blood cell life-span was normal in 12 other cases. Kinetic studies can therefore be useful, by demonstrating the mechanism of cytopenia observed after transplantation, and by facilitating the choice of appropriate treatment. PMID- 9395899 TI - Studies of T cell reconstitution after hematopoietic stem cell transplant. AB - Hematopoietic stem cell (HSC) transplantation, whatever its conditions, is associated with an increased risk of infections and tumoral complications, because of a delayed immune reconstitution. T-cell regeneration has been mostly investigated and appears to come more from graft and/or host mature T-cells, rather than from the differentiation/maturation of reinfused progenitors. In allogeneic setting, the immune defect is enhanced by the immune host/donor conflict and the use of prophylactic or curative immunosuppressive therapy. The tools used for studying post-transplant immunity are the following: immunophenotyping (kinetics and alterations of lymphocyte subset reconstitution), functional studies of T cell proliferation, cytokine production, cytotoxicity and signal transduction, as well as studies of T cell repertoire diversity. The CD4/CD8 cell immunophenotyping might be enough for routine clinical evaluation, allowing an adapted prophylaxis of opportunistic infections in those immune suppressed patients, while functional assays might be useful to evaluate the persistence overtime of defects in immune reconstitution. These overall assays are useful both for basic and clinical research and allow better understanding in the mechanisms for T cell regeneration in the diverse types of HSC transplants performed nowadays particularly after graft of purified HSC where immune reconstitution remains a key question. PMID- 9395900 TI - Immune reconstitution after blood cell transplantation. AB - Immune clinical events and pattern of recovery of total lymphocytes, T cells and CD4+/CD8+ T cell subsets, B cells and NK cells, were analysed after allogeneic blood cell transplantation (BCT) for 12 months following transplant (n = 33). Results were compared with immune recovery after allogeneic bone marrow transplantation (BMT) (n = 15) and autologous blood cell transplantation (n = 19). Rates of acute GVHD were comparable in the two allotransplantation groups. Fourteen out of 17 evaluable alloBCT group patients have presented an extensive cGVHD. Total T cell, CD4+T cells and CD56+NK cell reconstitution were improved in alloBCT group vs alloBMT group. CD4+/CD8+ ratio evolution were improved in alloBCT group vs alloBMT and autoBCT groups. These results confirm that rapid immune recovery is associated with allogeneic blood stem cell transplantation. PMID- 9395901 TI - Immune reconstitution following transplantation of selected hematopoietic stem cells. AB - The use of selected stem cell grafts has recently entered the era of clinical application. Its potential interest lies in the decreased contamination of the graft by tumor cells in the autologous graft setting and in the depletion of T lymphocytes from the graft in the allogeneic situation. Very few and only phenotypic studies of immunologic recovery post selected stem cell transplant have been performed. Most studies conclude an absence of influence of stem cell selection on quantitative neutrophil, NK, B-lymphocyte and CD8+ T-lymphocyte recovery. CD4+ T-lymphocyte recovery seems delayed in half of the studies, particularly in the context of allogeneic unrelated donor or HLA-mismatched transplants and this could explain viral infections frequently noted in this situation. Additional follow-up and comparative studies with non selected grafts including functional tests of immunity will be required to better assess the relationship of immune recovery post graft to the age of the patient, the type of donor and HLA compatibility between donor and recipient, the underlying pathology and the number of reinfused stem cells and accessory cells. PMID- 9395902 TI - Potential and limitations of HSV-TK-transduced donor peripheral blood lymphocytes after allo-BMT. PMID- 9395903 TI - [Usefulness of DNA ploidy, AgNORs, PCNA and c-erbB-2 as predictors of prognosis in patients with renal cell carcinoma]. AB - Seventy one patients with renal cell carcinomas were examined for a variety of markers associated with tumor malignancy: nuclear DNA ploidy, AgNORs, PCNA and c erbB-2. Usefulness of the markers in predicting the prognosis was studied by analyzing the relationship between each of these markers and the prognosis of the patients with renal cell carcinomas. DNA ploidy was analyzed by flow cytometry. AgNORs were stained by the silver colloid method. PCNA and c-erbB-2 were detected by immunohistochemistry. In all the patients examined, DNA ploidy, AgNORs, PCNA and c-erbB-2 were significant predictors of the prognosis. Of the patients with grade 2 carcinomas, the survival rate was significantly higher in the patients with the PCNA-positive cells of lower than 35.0% than in those with the positive cells of more than 35.0%. The patients without the expression of c-erbB-2 exhibited a significantly higher survival rate than those with the expression. In the patients with grade 2 carcinomas, however, neither DNA ploidy nor AgNoRs was a significant predictor of the prognosis. These findings suggest that PCNA and c erbB-2 provide more accurate information than the others to understand the biological characteristics of the grade 2 carcinomas and are useful in predicting the prognosis of the patients with grade 2 renal cell carcinomas. PMID- 9395904 TI - [A clinical study of intermittent hydronephrosis]. AB - A clinical study was conducted on the intermittent hydronephrosis in children. Of 78 children with hydronephrosis due to ureteropelvic junction obstruction operated between 1991 and 1995, 5 had intermittent hydronephrosis. All 5 patients were boys between 6 months and 6 years of age. Presenting symptoms were intermittent flank pain and vomiting. Ultrasonography performed during pain attack demonstrated dilation of the renal pelvis in all patients. Diuretic renography demonstrated obstruction in 4 of the 5 children and deterioration of renal function on the affected side in 2. The cause of ureteropelvic junction obstruction was aberrant vessels in 2 cases, fibrous band in 1, ureteral kinking within adventitia in 1 and a ureteral polyp in 1. Postoperatively, all patients have been relieved from the pain. In summary, ultrasonography at the time of symptom attack as well as diuretic renography are useful for the diagnosis and observation of intermittent hydronephrosis. PMID- 9395905 TI - [A clinicopathological study on carcinoma of the renal pelvis and ureter]. AB - We investigated the clinicopathological features of 62 patients with transitional cell carcinoma of the renal pelvis and ureter. Four patients had been treated for bladder cancer. Among 58 patients without precedent bladder cancer, 6 had coexistent bladder cancer and bladder cancer subsequently developed in 13. The 5 year cause-specific survival rate was 33% in cases with coexistent bladder cancer, 75% in those with subsequent bladder cancer and 62% in patients without association of bladder cancer. Distant metastases were found in 23 of 62 (37%) cases, the most frequent site being lymph nodes. The site of the primary tumor (renal pelvis and/or ureter) and the pathological findings such as grade, stage, type of infiltration, venous and lymphatic invasion, were significantly correlated to cause-specific survival. Multivariate analysis showed the most influential factors to be the type of infiltration and the site of the primary tumor. Therefore, patients with INF beta or gamma tumors both in the renal pelvis and ureter had a poor prognosis. However, association of bladder cancer was not related to survival. PMID- 9395906 TI - Two types of micturitions of ileal neobladder. AB - Eight patients were evaluated clinically, radiologically, and urodynamically to determine the outcome of continent urinary diversion with ileal neobladder performed to treat the recurrent superficial bladder cancer after cystectomy with subcapsular prostatectomy. The mean age of the patients was 55.3 years. After descending dissection of the urinary bladder without ligation or dissection of Santorini's plexus, the prostate was cut to the bladder neck distally for 2 cm under the subcapular prostatectomy. One patient who had a short 3 cm intestinal segment between the pouch and the urethra, had severely prolonged micturition with peristalsis in this short segment, and required a re-operation. Micturition was good in the other seven patients, all with detubularized neobladder directly to the prostate capsule in anastomosis. Pressure flow studies performed on these seven patients revealed two types of micturition; "fast bladder" and "intermittent flow", the latter resembling detrusor sphincter dyssynergia. PMID- 9395907 TI - [Cystic renal cell carcinoma: report of four cases]. AB - We report 4 cases of cystic renal cell carcinoma (RCC), one of simple cystic type (case 2) and three of multilocular cystic type (case 1, 3 and 4). All cases were diagnosed preoperatively as malignant neoplasms on the basis of radiological examinations, including CT scan and angiography. Pathological examination revealed that intrinsic cystic growth was the probable cause in the three cases of multilocular cystic RCC, while the simple cystic case was probably caused by secondary cyst formation as a result of tumor necrosis. Radical nephrectomy was performed in cases 1, 2 and 4 and partial nephrectomy in case 3. We recommend nephron-sparing surgery as an option in the management of select cystic RCC, given that many cystic RCCs are low grade and enveloped by distinct pseudocapsules with fibrous tissues. PMID- 9395908 TI - [A case of renal cell carcinoma found after skin metastasis]. AB - A case of renal cell carcinoma, found after skin metastasis is presented. A 76 year-old woman came to our hospital complaining of two painless subcutaneous tumors in her left chest. Histopathological diagnosis was clear cell carcinoma. She underwent left nephrectomy and histopathological findings revealed renal cell carcinoma, alveolar type, clear cell subtype, grade 1. Lung metastasis was proved soon after the operation. We removed skin metastasis several times after the nephrectomy. PMID- 9395909 TI - [Mucinous adenocarcinoma of the renal pelvis: a case report]. AB - A rare case of primary mucinous adenocarcinoma of the renal pelvis is reported. A 76-year-old woman was admitted to our hospital because of right abdominal fullness. Physical examination revealed a melon-sized (22 cm in diameter) tumor located in the middle and lower right quadrant of the abdomen. Computed tomography and transabdominal sonography revealed hydronephrosis and a renal stone. Retrograde pyelography was impossible because of ureteral obstruction on the right side. A diagnosis of severe hydronephrosis was made and a right nephrectomy was performed. The kidney measured 24 x 14 cm in size and contained 1,500 ml of mucinous material. The histological diagnosis was mucinous adenocarcinoma of the renal pelvis. The patient has had neither recurrence nor metastasis for 2 years following postoperative chemotherapy. PMID- 9395910 TI - [Microscopic foci of urachal carcinoma in an incidentally detected urachal cyst: a case report]. AB - A 33-year-old man who had been treated for chronic prostatitis was diagnosed to have urachal cysts by transabdominal ultrasonography. Cystoscopy revealed protuberance at the dome of the bladder. Computerized tomography scan and magnetic resonance imaging showed the mass to be mostly cystic but partly solid. Resection of the urachal cysts and partial cystectomy were performed. Histopathologically, most cysts had a normal cylindrical epithelium with retention of mucinous substance. However, several small cysts contained epithelial cells resembling tubulo-villous adenoma and showing mitotic figures. This case was concluded as urachal carcinoma detected in its very early stage. PMID- 9395911 TI - [A case of locally advanced squamous cell carcinoma in bladder diverticulum successfully treated by bladder-preserving therapy]. AB - A 62-year-old man visited our hospital with asymptomatic gross hematuria. On cystoscopy, two diverticula were identified at the dome of the urinary bladder and one of them was packed with a tumor. Biopsy showed squamous cell carcinoma (SCC), G1, but malignancy was not detected in the rest of the vesical mucosa. We selected bladder-preserving therapy. Pathological diagnosis after partial cystectomy was SCC, G1 > G2, pT3b. Postoperatively, 3 courses of adjuvant chemotherapy with methotrexate, bleomycin and cisplatin were performed. The patient has remained free of recurrence for 4.5 years. PMID- 9395912 TI - [Small cell carcinoma of the urinary bladder: a case report and review of the Japanese literature]. AB - A 50-year-old man presented with asymptomatic gross hematuria which he had first noticed 3 months earlier. Clinical examinations revealed a non-papillary, broad based tumor on the left lateral wall of the urinary bladder with a clinical stage of T3N0M0. The pathological diagnosis of a transurethral biopsy tissue specimen was small cell carcinoma. Neoadjuvant intraarterial infusion chemotherapy using cisplatin and adriamycin was initially administered but proved to be ineffective. Thus, we performed a radical cystectomy. The tumor tissue was apparently homogenous and composed of small cells arranged in sheets and solid patterns, and was staged to be pT3bR1L2V0N0. An electron microscopic study confirmed small cell carcinoma with neurosecretory granules. Postoperatively, 4 courses of adjuvant chemotherapy consisting of cisplatin, etoposide and ifosfamide were administered. The patient is alive without any evidence of tumor recurrence 26 months after the operation. PMID- 9395913 TI - [Adenocarcinoma occurring 37 years after augmentation ileocystoplasty for tuberculous bladder atrophy: report of a case]. AB - A 55-year-old woman was admitted with urinary frequency. She had undergone augmentation ileocystoplasty due to tuberculous bladder atrophy 37 years previously. Cystoscopy revealed a tumor on the posterior wall which had been augmented with the ileum. Partial cystectomy and bladder reconstruction using a segment of ileum and ascending colon were performed. Gross inspection showed a 15 x 10 mm, papillary tumor on the ileal mucosa near the vesico-ileal anastomosis. Histologically, moderately differentiated adenocarcinoma infiltrating into the muscle layer was surrounded by the normal ileal mucosa. She has been free of recurrence for 2 years postoperatively. This is the 8th case of adenocarcinoma following augmentation ileocystoplasty reported in the Japanese literature. PMID- 9395914 TI - [Bladder hemangioma in a child: a case report]. AB - We report a case of bladder hemangioma in a child. An 8-year-old girl was referred to our hospital for the evaluation of painless hematuria. Neither pyuria nor bacteriuria was detected. She had no dysuria. Intravenous pyelography showed deformity of the bladder without dilation of the upper urinary tract. Voiding cystourethrography revealed a filling defect of the bladder. Reddish blue masses were seen cystoscopically at the right wall and the dome of the bladder. Partial cystectomy was performed. From the histological findings, the diagnosis was cavernous hemangioma of the urinary bladder. Her postoperative course was uneventful and no recurrence was seen at the cystoscopical examination 3 months postoperatively. This is the 15th case of bladder hemangioma in children reported in Japan. PMID- 9395915 TI - A case of prostate cancer presenting as a symptomatic abdominal mass. AB - An 80-year-old man presented to our hospital complaining of an abdominal mass. On physical examination, a hard fist-sized mass was noted in the right lower abdomen. Needle biopsy of the prostate and abdominal mass showed moderately differentiated adenocarcinoma. The chest roentgenogram revealed multiple lung metastases. Clinical diagnosis was T3N3M1, stage D2. Serum prostate specific antigen (PSA) level (8,600 ng/ml) normalized after 3 months of anti-androgen therapy. Lung metastases disappeared after 11 months, while the abdominal mass was reduced to 25% of the pretreatment size after followup of 30 months. PMID- 9395916 TI - [Prostatic endometrioid carcinoma: a case report]. AB - An 89-year-old man presented with urinary retention. Digital rectal examination was benign despite elevated serum prostate-specific antigen (PSA) level. Suprapubic prostatectomy was performed under the diagnosis of benign prostatic hyperplasia. Histopathological diagnosis was endometrioid carcinoma showing positive immunohistochemical staining for PSA. Postoperatively, estrogen was administered for 9 months. After 37 months, serum PSA level began to increase and a palpable nodule was detected on digital rectal examination. Biopsy showed coexistence of endometrioid carcinoma and acinar adenocarcinoma. Hormonal treatment was resumed and the disease has been well controlled for 65 months postoperatively. PMID- 9395917 TI - [Statistics of the operation at Division of Urology, Shimada Municipal Hospital: 1992-1996]. PMID- 9395918 TI - High affinity human antibodies by phage display. AB - The development of phage display has now made it possible to consider the isolation of human antibodies directly without immunization. Recent advances in the field of human immunogenetics and in phage technology have led to the assembly of 'naive' human repertoires in vitro whose complexity approach that of the natural immune system. Screening of these libraries has allowed the isolation in one step of antibodies with affinities in the nanomolar range. PMID- 9395919 TI - Chimeric immunoglobulin E reactive with tumor-associated antigen activates human Fc epsilon RI bearing cells. AB - Crosslinking of immunoglobulin E molecules that are bound to the Fc epsilon receptors expressed on mast cells or basophils triggers activation of these cells, resulting in the development of a type I hypersensitivity. Targeting this potent immune reaction towards tumors by using IgE that reacts with a tumor associated antigen, may induce a local inflammation at the tumor site, and may therefore promote tumor regression. We have previously shown that murine IgE bound to tumor cells can activate murine mast cells to release TNF-alpha and histamine. To further investigate the therapeutic potential of IgE-mediated immunotherapy of carcinomas, we have developed human/murine chimeric versions, containing the murine variable regions and human constant regions, of both G250 and 323/A3 IgE. These chimeric IgEs are reactive respectively with the G250 renal cell carcinoma antigen and the Ep-CAM molecule, which is highly expressed by most carcinomas. Transfection of the respective chimeric heavy and light chain genes into recipient Sp2/0 myeloma cells yielded chimeric IgE-producing clones. Chimeric G250 and 323/A3 IgE reacted with tumor cells expressing the G250 antigen or Ep-CAM, respectively. To generate a cell line that expresses Fc receptors for human or chimeric IgE, the rat basophilic leukemia cell line RBL-7 was transfected with the human Fc epsilon RI alpha chain (RBL-7TZ) and subsequently tested for binding of chimeric IgE. Functional assays showed that both chimeric IgEs activated RBL-7TZ cells to release TNF-alpha when cultured with tumor cells that express the respective specific antigen. Furthermore, both chimeric IgEs were able to activate freshly isolated human basophils. PMID- 9395920 TI - Optimisation of human anti-tetanus toxoid antibody responses and location of human cells in SCID mice transplanted with human peripheral blood leucocytes. AB - A strategy for the production of human antibodies to tetanus toxoid (TT) is described. Human peripheral blood lymphocytes (PBL) were injected into severe combined immunodeficient (SCID) mice (termed hu-PBL-SCID) and the mice subsequently immunised with purified TT. By using low immunising doses of antigen, PHA activated PBL and PBL from donors who were recently immunised with TT, we established ongoing antibody responses to TT with specific recall IgG responses of up to 22 IU ml-1 in the murine plasma, which was greater than that in the donors' serum. Total IgG concentrations of up to 6 mg ml-1 were detected over a 32 week period. Lower levels of IgM and IgM anti-TT were also detected over this time. Large cellular infiltrations of human CD45+ and CD20+ cells were detected by immunocytochemistry in the mesenteric membranes, mesenteric lymph nodes and the pancreas 5 weeks after PBL were engrafted into a SCID mouse. Human cells were also observed in the lungs, liver, thymus and spleen. Cells isolated from the tissues were cultured with Epstein-Barr virus and the resulting B-cell lines produced Ig in vitro up to 7 weeks, with IgG and IgM anti-TT detected transiently in a culture of cells from the mesenteric membranes. PMID- 9395921 TI - Elevated expression levels of the 14-3-3 family of proteins in lung cancer tissues. AB - Immunochemical staining of lung cancer sections with a murine monoclonal anti-14 3-3 antibody showed a sharp discrimination of the cancer tissue from neighboring normal counterparts in 88 of 121 primary lung cancer tissue specimens of all four major lung cancer histologies; specifically, 32 of 48 adenocarcinomas, 36 of 44 squamous cell carcinomas, 10 of 13 large cell carcinomas, and 10 of 16 small cell carcinomas, respectively, were stained positively. Sets of the 10,000 x g supernatants of normal and cancerous lung tissue homogenates, each set prepared from surgically dissected tissues of the cancer and its surrounding normal part, were assayed for 14-3-3 proteins by the sandwich enzyme-linked immunosorbent assay using two different monoclonal antibodies to 14-3-3 proteins. The results of the assay demonstrated 7.2 times higher 14-3-3 protein content in the lung cancer tissue (378 +/- 200 ng ml-1) as compared with the normal lung (54 +/- 35 ng ml-1). These results indicate that the 14-3-3 family of proteins can be an effective marker for lung cancer diagnosis such as sputum cytodiagnosis and that 14-3-3 proteins might be involved in the development of lung cancers. PMID- 9395922 TI - Cytokeratin 8 and 19 as antigens recognized by adenocarcinoma-reactive human monoclonal antibody AE6F4. AB - The human monoclonal antibody (MAb) AE6F4 is secreted by a human-human hybridoma line established from the in vitro immunization of normal human peripheral blood lymphocytes with the human lung adenocarcinoma cell line, A549. This MAb is strongly reactive to lung cancer tissues. In the previous study, the antigens recognized by the MAb AE6F4 were purified from A549 cells and identified as 14-3 3 protein and 31 kDa cytosolic phospholipase A2 (cPLA2). The MAb AE6F4 also binds two kinds of antigens (53 kDa and 40 kDa), which are not related to 14-3-3 protein or 31 kDa cPLA2, in the human breast adenocarcinoma cell line, MCF-7. We purified a 38 kDa antigen, which is a degradation product of 53 kDa antigen from breast adenocarcinoma MCF-7 cells using ion-exchange and hydroxyapatite column chromatography. Two partial amino acid sequences of the purified 38 kDa antigen showed 95-100% homology to human cytokeratin 8 (CK8). Two-dimensional gel electrophoresis and immunoblot analysis of intermediate filament fraction separated from MCF-7 cells demonstrated that the 53 kDa and 40 kDa antigens were CK8 and CK19, respectively. Antigenic determinants on CK8 and CK19 recognized by the MAb AE6F4 were resistant to sodium periodate treatment, although antigenic determinant on 31 kDa antigen (14-3-3 protein and(or) cPLA2) was sensitive to this treatment. These results suggest that the MAb AE6F4 reacts with both carbohydrate and peptide antigenic determinants. PMID- 9395923 TI - Age differences in information-seeking among cancer patients. AB - Studies examining patient populations have found that information-seeking decreases with age. However, researchers usually define information-seeking as involving only the medical establishment, while they neglect other sources of information. The present study examined the use of two types of information sources, non-medical establishment (newspaper, television, and friends) and medical establishment (doctors and nurses), among seventy-five cancer patients aged eighteen to eighty-one years. Patients responded to questionnaires asking about information-seeking, desire for more cancer information, self-perception of their knowledge about cancer, and actual knowledge of facts about cancer. For the medical establishment source, information-seeking decreased with age; however, no age differences existed for seeking non-medical establishment information. In individuals with high levels of desire for information, older adults reported more information-seeking from non-medical sources than did younger adults. PMID- 9395924 TI - Relation between fluid intelligence and frontal lobe functioning in older adults. AB - This study reports the relations among normal aging, intelligence, and frontal lobe functioning. Intelligence tasks and frontal lobe functioning tasks were administered to 107 adults from two age groups (25 to 46 years and 70 to 99 years). Intelligence measures were assessed with two crystallized tests (WAIS Vocabulary and Information subtests), one fluid intelligence test (Cattell's Matrices), and one mixed, crystallized and fluid test (WAIS Similarities subtest). Frontal functioning was assessed using the Wisconsin Card Sorting Test (WCST) and two tests of verbal fluency. Significant age differences in favor of the young were found on the two intelligence tests with a fluid component and on all measures of frontal lobe functioning. Correlational analyses examining the relationship of intelligence measures to frontal variables indicated that these last measures were significantly correlated with only fluid intelligence tests in the elderly group. The implications for the relations among aging, fluid intelligence, and frontal lobe functioning are discussed. PMID- 9395925 TI - An exploration of attachment styles and personality traits in caregiving for dementia patients. AB - The present study examined the influence of caregivers' Attachment Styles (Anxious-ambivalent and Avoidant factors) and personality traits (Neuroticism, Extraversion, Agreeableness, and Conscientiousness) on their experiences of caring for dementia dependents. A total of 126 caregiver-dependent pairs participated in the study. Support was found for the contribution of the attachment style factors in explaining aspects of caregiver experiences. Those who chose to institutionalize dependents were higher on the Avoidance factor than those choosing to maintain them in the community. Less Anxious-ambivalent caregivers reported larger social support networks, and more satisfaction with the support received than those lower on this factor. The caregiver Anxious/ambivalence and Neuroticism dimensions seemed to function as generalized responses reflected in perceptions and appraisals of the stressful situation. PMID- 9395926 TI - Age differences in self-appraisal motivation. AB - One hundred and eight young (mean age 29 years) and 108 older adults (mean age 69 years) participated in a laboratory investigation of self-appraisal motivation. Participants were recruited via media advertisement to take part in a study of two novel and different thinking abilities and randomly assigned to either a similar others, dissimilar others, or temporal-self comparison referent condition. Each participant was administered two tests purported to measure different thinking abilities and received experimenter-controlled test feedback intended to manipulate participants' level of uncertainty about these abilities. Motivation to self-appraise was assessed via behavioral choice measures collected following the inducement of uncertainty about ability status. Results indicated that older adults were less likely than the young to initiate behaviors that would reduce uncertainty about ability. Subsequent post-hoc analyses suggested that self-appraisal motivation in young adulthood is not moderated by level of perceived efficacy, while in later adulthood an attenuation of self-appraisal motivation occurs as a result of low efficacy or heightened uncertainty about one's capabilities. PMID- 9395927 TI - An examination of the therapeutic benefits of focus groups on elderly worriers. AB - This study examines the effects on elderly worriers, of a focus group discussion about the topic of worry. All subjects (N = 21) were self-designated worriers, and at least seventy years of age. Pretest and-Posttest measures included questionnaires on worry, life satisfaction, and psychological symptom domains unrelated to the focus group topic. The percentage of the day spent worrying variable, which was the criterion variable for admittance into the groups, showed a significant reduction from pre to post. The focus group participants also evaluated the focus group experience as positive and beneficial. The value of focus groups for therapeutic effectiveness and data collection with the elderly are discussed. PMID- 9395928 TI - Appreciating background and culture: the South Asian elderly and mental health. AB - In the next decade there will be an increase in the number of elderly people from a South Asian background. All too often minority groups are treated as homogeneous, leading to inappropriate generalisations, unmet need, and unsuitable treatment and management. In order to understand and manage a person's illness it is necessary to appreciate the effects of their culture, experiences and environment. The South Asian community is well established in the UK and the attitudes of the growing elderly population towards mental illness, their expressions of distress, and views on management and treatment are only now being canvassed. Awareness of these issues is essential before epidemiological studies of depression and dementia and use of health services by this group will provide beneficial results. PMID- 9395929 TI - The Caregiver Activity Survey (CAS): development and validation of a new measure for caregivers of persons with Alzheimer's disease. AB - BACKGROUND: Most instruments that measure the impairments associated with Alzheimer's disease assess symptom severity. Little attention has been paid to the illness's impact on the time formal and informal caregivers spend caring for Alzheimer's individuals. A tool that measures the time spent caregiving would help to determine the economic impact of the illness. The Caregiver Activity Survey (CAS) was developed to measure the time caregivers spend aiding Alzheimer's patients with their day-to-day activities. METHODS: The test-retest reliability of the CAS was assessed during a 3-week study with 42 Alzheimer's patients and their caregivers. The CAS was validated with the Alzheimer's Disease Assessment Scale Cognitive Subscale (ADAS-Cog), the Mini Mental State Exam (MMSE) and the Physical Self Maintenance Scale (PSMS). RESULTS: The final version of the CAS consists of six items (communicating with the person, using transportation, eating, dressing, looking after one's appearance and supervising the person). The six-item CAS total score has high test-retest reliability, with ICC = 0.88 between weeks 1 and 3. The scale has strong convergent validity with the ADAS-Cog (r = 0.61), MMSE (r = -0.57) and PSMS (r = 0.43). Efforts to include a dimension that reflects caregiver burden were not successful, in part due to the reluctance of caregivers to acknowledge that caregiving is bothersome. CONCLUSIONS: The CAS provides a new tool that measures time spent caring for Alzheimer's individuals. The instrument may be used to augment existing clinical assessments that measure the efficacy of potentially therapeutic agents for persons with Alzheimer's disease. PMID- 9395930 TI - Psychogeriatrics in South-East Asia. AB - A common phenomenon in South-East Asia is ageing of the population. This article describes the various stages of development of psychogeriatrics in Hong Kong, Singapore, Malaysia, Thailand, Indonesia and the Philippines. It is only in the last few years that more systematic development of psychogeriatric services has begun under the pressure of an ageing population. The model of service delivery in Hong Kong can serve as an example of development of psychogeriatric services in South-East Asia. PMID- 9395931 TI - Cognitive impairment and social distress as different pathways to depression in the elderly: a cross-sectional study. AB - BACKGROUND: This study investigates the recent suggestion that some putative aetiological factors for depression, such as cerebral deterioration and social distress, may act differentially in the aetiology of depression in old age. METHOD: In a cross-sectional study, a community sample of 654 elderly subjects were interviewed with Short-CARE to assess the prevalence of depression and cognitive impairment. Information was collected for a variety of potential risk factors for depression such as exposure to social support deficit, threatening life events, impairment, disability and handicap. RESULTS: The prevalence of depression was 17% and that of a broad concept of cognitive impairment 23.9%. This analysis found associations between depression and exposure to social support deficits and threatening life events in the year prior to interview. These associations were considerably stronger for those subjects with no cognitive impairment than for those with cognitive impairment. We also found a progressive lowering in the strength of these associations the higher the chance of cognitive impairment measured as a longitudinal variable using both the Dementia Diagnostic Scale (DDS) and the Organic Brain Syndrome Scale (OBS) included in Short-CARE. CONCLUSIONS: The results of this theory-driven analysis lend some support to the notion of at least two differential pathways to depression in the elderly, one via social distress factors and another mediated by cerebral deterioration clinically expressed as cognitive impairment. PMID- 9395932 TI - Cognitive impairment in geriatric chronic schizophrenic patients: a cross national study in New York and London. AB - Severe cognitive impairment has been reported in large numbers of geriatric chronic schizophrenic patients in the US, with this impairment also being related to severe negative symptoms and adaptive deficits. It is not clear if this impairment is related to the particular environment of the American state hospitals and would not generalize to other countries. In this study, a sample composed of geriatric (age > 70) chronic schizophrenic patients in London, who were assessed by the Team for Assessment of Psychiatric Services (TAPS) (N = 137), and a group of geriatric chronic schizophrenic patients in a New York psychiatric center (N = 86) were compared for the severity of cognitive impairment and on measures of adaptive functioning. Patients received essentially identical Mini-Mental State Examination (MMSE) scores, but differed on 3/4 measures of adaptive functioning. The correlations among all four aspects of adaptive deficit and MMSE scores were very similar in the two samples, suggesting that cognitive deficits and their relationship with adaptive impairments are relatively invariant across different psychiatric systems of care, while adaptive functioning deficits are more variable and possibly more influenced by environmental factors. These data add to previous results suggesting that cognitive impairment is a common feature in poor outcome geriatric patients with schizophrenia. PMID- 9395933 TI - Age- and education-specific reference values for the Mini-Mental and modified Mini-Mental State Examinations derived from a non-demented elderly population. AB - MAIN OBJECTIVE: To report age- and education-specific reference values for the Mini-Mental State Examination (MMSE) and Modified Mini-Mental State (3MS) Examination. DESIGN: Cross-sectional study. SETTING: Community and institutional settings in five regions across Canada. PARTICIPANTS: 7754 subjects aged 65 and over randomly chosen to take part in the Canadian Study of Health and Aging. Subjects classified as cognitively impaired or demented following a clinical and neuropsychological examination were excluded. MEASUREMENTS: Total scores on the MMSE and 3MS, and the degree to which they are influenced by the age, sex, education, mother tongue and living environment of the subject. RESULTS: Reference values on the two tests are reported through various descriptive statistics for five age groups and four education levels. These values decrease with age and increase with years of schooling. Test scores are also influenced by the subject's sex and mother tongue, albeit to a lesser extent. These observations led to the development of predictive equations of the performance to be expected from a 'normal' elderly subject, given his/her socio-demographic characteristics. CONCLUSION: The use of the reference values and related predictive equations will allow the clinician to interpret a patient's performance on two widely used cognitive tests, in light of the value expected from a group of 'normal' subjects with the same sociodemographic profile. PMID- 9395934 TI - Clinical diagnoses and disability of cognitively impaired older persons as predictors of stress in their carers. AB - BACKGROUND: Aspects of the caring relationship are often promoted as more important than the clinical features of the care recipient in predicting caregiver wellbeing. However, studies of consequences of caring for cognitively impaired people seldom include detailed measures of the diagnostic profile and disability of the care recipient. METHODS: Ninety community-living elderly persons with cognitive impairment were clinically assessed for severity on a range of illnesses. Their disability was examined via informant reports. Informants (88% of whom were primary carers) provided information on the behaviour and personality of the subject and reports of their own (informant) wellbeing. Using multiple regression, features of the subjects' clinical profile (severity of diseases, disability, behavioural problems and personality change) were examined as predictors of informant wellbeing. After controlling for subject clinical profile, we explored the additional associations between informant stress measures and other descriptors of the subject, caregiver and their relationship. RESULTS: The subjects' clinical characteristics, in particular disability and disturbed behaviour, were strong predictors of caregiver wellbeing, accounting for most of the explained variance. After control for the subjects' clinical profile, few of the sociodemographic, caregiver or relationship variables examined had any influence on caregiver outcome measures. The exceptions were caregiver time demands, older subject age and self identification as primary carer. Coresidence was not associated with caregiver distress. CONCLUSION: Clinical characteristics of the care recipient are determinants of caregiver wellbeing, while socio-demographic, caregiver and relationship characteristics are less influential. PMID- 9395935 TI - Experience with a Swedish version of the Geriatric Depression Scale in primary care centres. AB - OBJECTIVE: The purpose of this study was to use a Swedish version of the Geriatric Depression Scale (GDS-20) for diagnosis of depression in the elderly in primary care. DESIGN: Elderly consecutive patients visiting two primary care centres (> or = 65 years of age; N = 1189) were rated by educated nurses using the GDS-20. SETTING: All elderly patients attending two primary care centres in an urban-based community in the south of Sweden. PATIENTS: Of the 1189 patients interviewed, 1002 were rated using the GDS-20. MEASURES: The GDS-20, and in 26 patients also the Geriatric Mental State Schedule--Depression Scale (GMSS-DS). RESULTS: Of 1002 rated patients, 93 had scores of 5 or above on the GDS-20. Further analysis showed that 158 (13.3%) suffered from affective disorders. CONCLUSION: Depression in the elderly is underdiagnosed in primary care centres. A screening instrument such as the GDS-20 is of value in identifying the patients. PMID- 9395936 TI - Concurrent validity of a telephone-administered version of the Gospel Oak instrument (including the SHORT-CARE). AB - OBJECTIVE: The aim of the study was to establish the concurrent validity of a telephone-administered version of the survey measures utilized in the Gospel Oak studies, the core of which was the SHORT-CARE. DESIGN: Comparisons were made between data obtained by administering the interview in its conventional, face-to face form with those generated by conducting it by telephone. SETTING: A UK teaching hospital. PATIENTS: Elderly subjects of both sexes, recruited from geriatric and psychogeriatric day hospitals. MEASURES: The Depression Diagnostic Scale, Dementia Diagnostic Scale and Organic Brain Syndrome scale (all taken from the SHORT-CARE) and the London Handicap Scale. RESULTS: For depressive symptomatology, cognitive impairment, subjective memory impairment and handicap, intraclass correlations were 0.86, 0.89, 0.83 and 0.70, respectively. The kappa coefficient for agreement on case-level diagnosis of pervasive depression was 0.79. CONCLUSIONS: These results indicate that the instrument is broadly reliable when administered by telephone. PMID- 9395937 TI - Cognitive function in community-dwelling elderly with chronic medical conditions. AB - OBJECTIVES: The aim of this study was to examine the effect of chronic medical conditions on cognitive function in a sample of community-dwelling elderly (N = 4528). METHODS: A checklist of 18 chronic medical conditions was used to determine whether respondents were suffering from specific disease states. The Mini Mental Status Examination (MMSE) was administered to assess cognitive functioning. RESULTS: Statistically controlling for the effects of age, education and depression, respondents with asthma/bronchitis and stroke had a tendency to perform worse on the MMSE than those without these conditions. None of the 18 medical conditions was associated with a greater proportion of respondents scoring below the cutoff for cognitive dysfunction. CONCLUSION: It appears that- with the possible exception of stroke and asthma/bronchitis-cognitive function in community-dwelling elderly is not consistently affected by specific disease states. PMID- 9395938 TI - Ethical issues. PMID- 9395939 TI - Depression in old age: questions concerning prevalence studies. PMID- 9395940 TI - Current awareness in geriatric psychiatry. PMID- 9395941 TI - Reperfusion injury: fact or myth? PMID- 9395942 TI - Repair of coarctation of the aorta in neonates and young infants. AB - In repair of coarctation in neonates or young infants, inadequate removal of ductal tissue, failure to address hypoplasia of the aortic arch, and suture line tension have been reported to be important factors of residual or early recurrent stenosis at the coarctation repair site. In a consecutive series of neonates and young infants with coarctation, who were all operated without delay with extended resection, the clinical outcome regarding the development of restenosis and hypertension was studied. In addition, the resected specimens were investigated regarding the completeness of resection of ductal tissue. Twenty-five consecutive neonates and young infants (median age 22 days, range 5 to 39 days) who underwent surgical correction of coarctation were reviewed; the resected specimens were examined histologically to document the extent of ductal tissue in the aortic wall. Fifteen patients had a preductal coarctation with associated cardiovascular anomalies including a hypoplastic aortic arch (n = 11). The remaining 10 patients had a paraductal coarctation without associated intracardiac anomalies. In all patients, the isthmus was bypassed and an end-to-side anastomosis was constructed between the descending aorta and the undersurface of the proximal aortic arch (n = 13) or the distal ascending aorta (n = 12). In 13 patients without marked hypoplasia of the aortic arch, the coarctation repair was performed through a left thoracotomy. In the remaining 12 patients, the coarctation was repaired through a median sternotomy with CPB and hypothermic circulatory arrest, on the basis of an associated hypoplastic aortic arch (n = 4), hypoplastic aortic arch with intracardiac anomalies (n = 7), or a "bovine" innominate artery (n = 1). There was no perioperative or late mortality. At a median follow-up of 15 months, 1 patient (4%) developed a recurrent stenosis at the coarctation repair site; in the remaining 24 patients, echocardiography showed a widely patent anastomosis with no evidence of a hemodynamically significant gradient. None of the patients had hypertension. Histologic examination of the resected specimens demonstrated the presence of ductal tissue in the descending aorta with maximal extension into its lateral wall (mean 5.2 mm). In all specimens of the paraductal subtype, there was also extension of ductal tissue into the lateral wall of tbe isthmus (mean 3.9 mm). We conclude that: (1) in the absence of marked hypoplasia of the proximal aortic arch, coarctation can be repaired with low mortality and morbidity via a left thoracotomy; (2) in the presence of marked hypoplasia of the proximal aortic arch and/or if associated intracardiac defects also need to be repaired, we advocate repair of the coarctation and associated defects through a median sternotomy with circulatory arrest; (3) in view of the absence of postoperative hypertension in this series, early repair of aortic coarctation is recommended; and (4) because ductal tissue may extend not only into the descending aorta but also into the isthmus, complete excision of the coarctation and bypass of the isthmus are valuable techniques to avoid secondary constriction of the aorta by ductal tissue. PMID- 9395943 TI - Twenty-year follow-up of acute type a dissection: the incidence and extent of distal aortic disease using magnetic resonance imaging. AB - A persistent distal false lumen (PDFL) after surgical repair of type A aortic dissection is the most important factor in determining long-term survival. It has been suggested that changes in surgical technique reduce the incidence of distal false lumen. We report the findings of a 20-year follow-up (mean 5.2 years) on 87 patients who have undergone surgical repair of type A aortic dissection with all survivors undergoing magnetic resonance (MR) scanning of the entire aorta. Early mortality was 27.5%, and actuarial 5-, 10-, and 15-year survival was 65%, 28% and 20% respectively. Early mortality had decreased to 18% in the last 5 years. The most common cause of late death was related to distal aortic disease, accounting for 47% of all late deaths with a peak incidence at 7-10 years after surgery. The incidence of PDFL in survivors was 72%, despite the fact that 82% of all intimal tears were resected at time of operation. Incidence was not affected by extension of the repair into the aortic arch nor by the use of the open technique or Gelatin-Resorcine-Formal tissue glue. In patients with a distal false lumen 6% had reached a maximum aortic diameter of 6 cm in at least one plane on MR scanning and 25% had reached 5 cm. We conclude that if dissection has extended beyond the arch at time of presentation then the choice of surgical technique does not prevent the persistance of a distal false lumen. MR scanning gives ideal anatomical and functional assessment of distal aortic disease and provides the surgeon with all the necessary information to plan the timing and indications for further surgery. PMID- 9395944 TI - Endarterectomy of the ascending aorta: an alternative method in patients with extensively calcified (porcelain) aorta requiring aortic valve replacement. AB - A variety of surgical techniques have been described to manage the extensively calcified (porcelain) aortic root in patients requiring aortic valve replacement. We report two cases in which endarterectomy of the proximal ascending aorta was successfully performed. Endarterectomy of the left main coronary artery ostium was also performed in the one patient, and aortic root enlargement with patch aortoplasty in another. The technical aspects of the procedure and alternative approaches are discussed. PMID- 9395945 TI - Coronary artery bypass grafting in patients with chronic congestive heart failure: a 10-year experience with 203 patients. AB - From 1983 to 1992, 203 patients with chronic congestive heart failure and no angina underwent primary coronary artery bypass. This represented 3% of patients undergoing coronary artery bypass grafting. Ninety-two percent of the patients were in New York Heart Association (NYHA) functional class III or IV prior to undergoing coronary artery bypass grafting. Thallium perfusion imaging was performed in 21% of the patients, with a reversible defect present in 88%. An internal mammary artery graft was used in 70% of the patients. The hospital mortality was 6.0% and the actuarial survival at 5 years was 59%. An improvement in NYHA functional class occurred in 75% of the surviving patients with a mean improvement of 1.6 +/- 0.6 functional classes. Univariate analysis identified risk factors for hospital death as emergency operation, recent myocardial infarction (< 30 days), and the need for an intra-aortic balloon pump. A trend emerged for nonuse of an internal mammary artery to predict hospital death. A positive thallium perfusion scan was not a predictor of early or late survival, nor did it influence NYHA functional class. The use of the internal mammary artery significantly enhanced late survival (p = 0.01), however, did not affect the functional class of survivors. We conclude that coronary artery bypass grafting is effective in ameliorating symptoms of chronic congestive heart failure in patients suffering from chronic ischemic cardiomyopathy and can be performed with acceptable early and late mortality. PMID- 9395946 TI - Noninvasive nasal mask BiPAP management for prolonged respiratory failure following cardiovascular surgery. AB - The purpose of this study was to assess the efficacy of nasal mas bi-level positive airway pressure (BiPAP) support in managing respiratory failure following cardiovascular surgery. A total of 20 patients requiring postoperative prolonged respiratory support of 72 hours or longer were studied. BiPAP support was used for eight patients (BiPAP group); the other 12 patients were managed using ordinary oxygen mask treatment (control group). The mean age of the BiPAP group and control group was 65 and 58 years of age, respectively. The mean period of postoperative endotracheal intubation of the BiPAP group and control group was 12 +/- 5 days and 7 +/- 1 days, respectively. Reintubation was necessary in two patients of the control group. The BiPAP group patients required no reintubation. BiPAP support was discontinued within 48 hours in 6 out of 8 patients. The respiratory rates of control group increased (p < 0.1) 24 hours after extubation, however, the respiratory rates of the BiPAP group remained unchanged. The values of the respiratory index of the BiPAP group improved significantly (p < 0.01) after BiPAP management (from 1.5 +/- 0.2 to 0.9 +/- 0.2). The values of the control group, however, remained unchanged. A-aDO2 and Qs/Qt decreased (p < 0.1) in the BiPAP group. There were no significant differences in central venous pressure or circulatory status between the two groups. In conclusion, BiPAP support is a noninvasive management technique for postoperative respiratory failure and may also prevent prolonged endotracheal intubation. PMID- 9395947 TI - Pulmonary valve reconstruction in absent pulmonary valve syndrome: a new technique. AB - BACKGROUND: In patients with absent pulmonary valve syndrome, the relief of the pulmonary regurgitation at the time of primary repair improves both the early and late results. Though homograft and heterograft valves and conduits have been used for this purpose, both are not easily available and are known for late failure. Monocusp and bicuspid semilunar valves made out of pericardium have their own problems. Hence, a technique of reconstructing an autologous competent 3-cusp valve from the native tissues was developed. METHODS: Two posterolateral semilunar cusps were fashioned from the anterior wall of the main pulmonary artery. The anterior cusp was made from autologous pericardium stitched to the autologous pericardial patch used to widen the right ventricular outflow tract. RESULTS: This method of reconstruction was used in two patients aged 9 and 22 years, respectively. Visual assessment and passive testing after reconstruction revealed well functioning neopulmonary valves in both patients. The second patient, who had an unevenful hospital course, showed only mild pulmonary regurgitation at 5 years postreconstruction. CONCLUSIONS: As 2 of the 3 cusps are fashioned from the pulmonary arterial wall as a pedicled graft, we believe that they will retain their viability and grow with the pulmonary artery. Simultaneous reduction in the size of the pulmonary arteries will relieve bronchial compression when present. The anterior pericardial cusp, even if it eventually shrivels up, is unlikely to produce serious hemodynamic derangements. PMID- 9395948 TI - A new instrument for immobilization and hemostasis during minimally invasive direct coronary artery bypass ("MIDCAB doughnut"): experimental study. AB - A new instrument for the immobilization and hemostasis of an anastomotic site is described for use during minimally invasive off-pump direct coronary artery bypass (MIDCAB). The mechanism is based on air suction of the heart surface. The instrument is unique in that it can avoid direct temporary of the distal end of the coronary artery to control bleeding. With this instrument, anastomosis of left internal thoracic artery to left anterior descending artery (LAD) was successfully performed on three pigs. Additionally, in six patients the LAD could be stably and securely immobilized in the beating heart by the present instrument. The instrument is hereinafter referred as "MIDCAB doughnut," which can make an operative field motionless and bloodless without distal snaring. PMID- 9395949 TI - Influence of two blood conservation techniques (cardiotomy reservoir versus cell saver) on biocompatibility of the heparin coated cardiopulmonary bypass circuit during coronary revascularization surgery. AB - Blood conservation during cardiac surgery is critically important because of the inherent risks in homologous blood transfusions. Two techniques for the intraoperative conservation of blood--retransfusion of the red cells using a cell saver (CS), or retransfusion of the blood using a cardiotomy suction (CTR) system -were compared using biocompatibility markers, granulocyte activation, and production of oxygen-free radicals (OFR). In the CTR group, heparin coated circuits with an uncoated cardiotomy reservoir were used. For the CS group, identical heparin coated cardiopulmonary bypass (CPB) sets, without a cardiotomy reservoir but with a CS, were used. In each group, eight patients had coronary artery bypass grafting performed. The capacity of the whole blood and the granulocytes to produce OFR was estimated by a chemiluminescence, and granulocyte activation was measured as release of the granulocyte granule proteins myeloperoxidase (MPO) and lactoferrin. A significantly reduced capacity to produce OFR by the whole blood was noted at 45 minutes of CPB in the CTR group (68% +/- 17% vs 94% +/- 16% in the CS group). MPO release was higher after 3 hours (p = 0.05) and 20 hours (p < 0.05), postoperatively, in the CTR group (417 +/- 77 micrograms/L and 257 +/- 31 micrograms/L vs 246 +/- 25 micrograms/L and 164 +/- 12 micrograms/L, respectively, in the CS group). We conclude that the heparin coated CPB circuit with the uncoated cardiotomy reservoir may be less biocompatible than the identical CPB set used together with the CS. PMID- 9395950 TI - Jarcho-Levin syndrome associated with atrial septal defect and partial anomalous pulmonary venous return: a case report. AB - A 61-year-old woman suffering from Jarcho-Levin syndrome (JLS) was associated with atrial septal defect and partial anomalous pulmonary venous return and underwent corrective surgery. Pressure controlled postoperative ventilator therapy is preferred in patients with JLS. PMID- 9395951 TI - Simple method to improve the effectiveness of brain retrograde perfusion during total circulatory arrest. PMID- 9395952 TI - Valvular closure of atrial septal defect and its extended use. PMID- 9395953 TI - [Measurement of plaque volume using three-dimensional intravascular ultrasound: in vitro study]. AB - The usefulness of three-dimensional echocardiography using intravascular ultrasound (3D-IVUS) for the measurement of plaque volume was evaluated by comparing plaque volume derived from 3D-IVUS with that directly measured in 10 autopsied iliac or femoral plaque models (5-15 mm long). Using IVUS (3.5 F, 30 MHz), sequential cross-sectional images for three-dimensional datasets were acquired with a motorized catheter pullback device connected to the three dimensional reconstruction system. Three-dimensional reconstruction was performed from the sum of the two-dimensional cross-sectional views. Plaque volumes were calculated using a summation of disks algorithm based on the reconstructed multiple short-axis cross-sections from the three-dimensional data. Three dimensional IVUS demonstrated a good correlation with direct measurement of plaque volume (y = 0.71x + 0.001, r = 0.80, SEE = 0.003 ml), so is useful for the measurement of plaque volumes in the experimental models. PMID- 9395954 TI - Hemodynamic effects of warm bathing in a Hubbard tank and exercise loading in patients after myocardial infarction. AB - Hemodynamic parameters were measured during bathing and exercise testing in 43 patients with myocardial infarction (mean age: 60.2 years) to investigate the predictive parameters to determine when patients could safely resume bathing. Patients took a fresh water bath at 42 degrees C in the supine position for 5 min in a Hubbard tank. Group A showed an elevation of pulmonary capillary wedge pressure (PCWP) during bathing of 10 mmHg or more (23 patients, mean age: 61.7 years) and group B showed an elevation of less than 10 mmHg (20 patients, mean age: 60.5 years). Continuous multistep exercise tests were performed with a bicycle ergometer in the supine position, and hemodynamic parameters were measured at up to 50 W for 3 min on the day before the warm bathing test. There were no significant differences in the changes of arterial pressure and heart rate between the two groups. The PCWP at 3 min with a load of 50 W was significantly higher in group A (26.9 +/- 9.0 mmHg) than in group B (16.7 +/- 9.1 mmHg, p < 0.01). The stroke index (SI) during exercise testing was significantly lower in group A than in group B. The difference in the stroke index from baseline values (delta SI) at 3 min with a load of 50 W was significantly lower in group A (3.5 +/- 5.5 ml/m2/beat) than in group B (10.6 +/- 7.0 ml/m2/beat, p < 0.01). Similarly, delta CI and delta oxygen pulse during testing were significantly lower in group A than in group B. The physical work capacity and ejection fraction of the left ventricle of group A were significantly lower than those of group B, whereas the left ventricular end-diastolic pressure was higher in group A than in group B. CI, delta CI, SI, delta SI, METs, oxygen pulse, and delta oxygen pulse were examined by regression analysis and multivariate analysis to predict a significant elevation of delta PCWP during bathing. delta SI (p = 0.0032), delta CI (p = 0.0094), delta SI + METs (p = 0.0051), delta CI + METs (p = 0.0061), delta CI + delta SI (p = 0.0084), and delta CI + delta SI + METs (p = 0.0093) showed the highest correlations with delta PCWP. These findings suggest that changes in delta CI, delta SI, and METs are good predictive parameters for determining when patients may safely resume bathing. We suggest that patients with myocardial infarction, reduced cardiac function and a physical work capacity of approximately 4.0 METs, delta SI: 5 ml/m2/beat and delta CI: 2.4 l/min/m2 resume bathing only after careful consideration. PMID- 9395955 TI - [Mechanism of increase in exercise tolerance in patients with acute myocardial infarction]. AB - The contribution of cardiac output reserve and the skeletal muscle to exercise capacity was investigated in 24 patients with acute myocardial infarction. Symptom-limited exercise tests with a cycle ergometer were performed at 1 week, 3 weeks, and 3 months after the onset of the first infarction. Ventilatory gas was analyzed throughout the testing, and peak oxygen uptake (peak VO2) and anaerobic threshold (AT) were determined. During the test, the cardiac index (CI) was measured by the dye dilution method and the change in CI during exercise (delta CI) was calculated as an index of cardiac output reserve. The cross-sectional area of the thigh muscles (CSA) at the level of 10 cm above the patella was measured using computed tomography. Peak VO2 and AT increased significantly from 1 week to 3 months after the onset of infarction. delta CI increased significantly from 1 week to 3 weeks, and CSA increased significantly from 3 weeks to 3 months. Peak VO2 correlated significantly with both delta CI and CSA at each measurement point, as was AT with delta CI and CSA. Change in peak VO2 correlated with change of delta CI from 1 week to 3 weeks, and also with both delta CI and CSA from 3 weeks to 3 months. These results suggest that both cardiac output reserve and peripheral factors contribute to the exercise capacity up to 3 months after the onset of myocardial infarction. In particular, peripheral factors such as muscle volume are important to improve exercise capacity from 3 weeks to 3 months. PMID- 9395956 TI - [Usefulness of magnetic resonance imaging for managing patients with prosthetic carbon valve in the mitral position]. AB - The safety, findings and clinical usefulness of magnetic resonance (MR) imaging were assessed in patients with a prosthetic carbon valve in the mitral position. In vitro deflection, heating and image distortion due to the magnetic field of a 1.5 tesla MR machine were examined in three carbon valves (CarboMedics, St. Jude Medical and Bjork-Shiley valves). In vivo MR imaging of the left ventricular horizontal long-axis, vertical long-axis and short-axis views was performed by electrocardiographically synchronized spin echo and field (gradient) echo techniques in eight patients with prosthetic mitral carbon valves, consisting of six CarboMedics valves, one St. Jude Medical valve and one Bjork-Shiley valve. No deflection and significant heating was seen in all three valves in vitro. Although little image distortion was shown in the CarboMedics and St. Jude Medical valves, a small distortion toward the frequency encoded direction was seen in the Bjork-Shiley valve but caused no difficulty in assessing the surrounding images. Four of the eight patients had normal sinus rhythm and the other four had atrial fibrillation. The prosthetic valves were depicted as signal voids in the images taken by both spin echo and field echo techniques in vivo. Clear structural information with little image distortion of the adjacent tissues of the prosthetic valves were obtained in all patients, although the image of the Bjork-Shiley valve which contained stainless steel in the frame had a slightly stronger distortion than those of the CarboMedics and St. Jude Medical valves which contained titanium. The stainless wire suture material used to close the sternal incision was depicted as a signal void, and the areas of the signal loss were larger in the images taken by the field echo technique than those by the spin echo technique. The images taken by the spin echo technique in patients with atrial fibrillation had reduced quality due to the irregularity of repetition time. Cine MR imaging by the field echo technique showed physiological mitral regurgitant jets as signal loss within the flowing blood, which appeared as high signal intensity, bidirectionally in the bileaflet mechanical valve and unidirectionally in the monoleaflet mechanical valve. An abnormal cavity was seen behind the basal left ventricular myocardium in one patient with a CarboMedics valve. The wall of the abnormal cavity was disrupted abruptly and the rest of the wall consisted of pericardium and adjacent tissue in the image taken by the spin echo technique. The image taken by the field echo technique showed an abnormal jet flow from the basal part of the left ventricular cavity into the abnormal cavity, which was compatible with left ventricular pseudoaneurysm. Two dimensional echocardiography and Doppler color flow mapping disclosed the abnormal cavity and the abnormal flow inside, but failed to show the connection between the left ventricle and the cavity due to reverberation of the ultrasound signal by the prosthetic valve. These findings suggest that MR imaging is a safe and promising method to assess the complications and valvular function in patients with a prosthetic carbon valve in the mitral position. PMID- 9395957 TI - Antegrade or retrograde catheterization across a ventricular septal defect. AB - The success rate and the most suitable catheter tip for crossing over various types of ventricular septal defects (VSDs) were examined as a preliminary study for transcatheter closure of VSD. The 18 consecutive patients with various types of VSD were aged from 1 to 95 (mean [+/- SD] 13 +/- 22) months. Body weight was 3.7 to 25.0 (8.0 +/- 5.1) kg. Two-dimensional echocardiography showed that the maximal diameter of the defects ranged from 1.0 to 12.0 (7.5 +/- 3.1) mm. There were 10 patients with perimembranous defects, 4 with outlet defects, 2 with muscular defects, and 2 with tetralogy of Fallot with perimembranous defects. An angiographic balloon catheter, or an original or modified Judkins right coronary catheter could be passed through the VSD antegradely or retrogradely in 16 of 18 patients. In only two patients, with a small VSD of 2.0 and 1.0 mm, the catheter could not cross over the defect. The catheter entered the ascending aorta antegradely in 12 cases. The Judkins right coronary catheter is most suitable for crossing a VSD either antegradely or retrogradely. PMID- 9395958 TI - [Evaluation of intramyocardial coronary flow velocity pattern before and after surgical repair of Bland-White-Garland syndrome by pulsed Doppler echocardiography]. AB - Anomalous origin of the left main coronary artery from the pulmonary artery (Bland-White-Garland syndrome) is a rare congenital anomaly. Intramyocardial coronary flow dynamics by pulsed Doppler echocardiography were studied in three patients with this syndrome who underwent surgical repair by Hamilton's method. Before surgery, the intramyocardial flow at the ventricular septum showed a retrograde velocity pattern which had two peaks in systole and diastole in all patients. After surgery, two patients with successful repair showed a biphasic intramyocardial flow pattern which consisted of retrograde and antegrade flows in systole and diastole, respectively. In contrast, one patient who had a residual shunt between the left coronary artery and the pulmonary artery showed a biphasic pattern which had antegrade flow in systole and retrograde flow in diastole. These results may suggest that the evaluation of postoperative intramyocardial coronary flow velocity pattern by pulsed Doppler echocardiography is useful for detecting a residual shunt after surgical repair of Bland-White-Garland syndrome. PMID- 9395959 TI - [Cardiovascular imaging in-a-month. A 60-year-old woman with systolic murmur after repeat mitral valve replacement]. PMID- 9395960 TI - The range of provider/insurer configurations. AB - In the past few years, health care providers have begun to seriously explore the possibilities of forming various kinds of provider/insurer entities. As most people in the health care industry now know, various states, as well as the federal government, have been working on one form or another of legislation or regulations that would permit health care providers to either (1) become insurers in their own right or (2) at least be able to contract more directly with consumers by taking on "full-risk" contracts from health maintenance organizations and insurance companies while, commensurably, assuming additional "delegated authorities" with respect to such contracts with minimal interference from state insurance departments. PMID- 9395961 TI - Physician equity in health care delivery systems: three alternative models. AB - The 1990s have seen many health care organizations attempting to merge, acquire, or affiliate with physician groups. Many have failed to provide physicians a stake in the success of the newly formed enterprise, frequently resulting in declining physician productivity, poor morale, and large operating losses. These problems warrant a reexamination of the traditional acquisition model of growth in favor of structures that retain a physician ownership component. This article examines three models of health care organization in which physicians share in the success of the enterprise and compares them in terms of ownership structure, governance, and funds flow. PMID- 9395962 TI - Doing it all in the public eye: a comparative analysis of California district hospital affiliations. AB - California contains 74 hospital districts, which are local governmental entities with publicly elected boards. Historically, they have been responsible for operating hospitals in remote areas to provide for the health care needs of their communities. Because the health care market has become increasingly competitive and because some of these once rural areas have grown into large cities, these districts are seeking partners for their hospitals in hopes of preserving their long-term financial viability. The terms reached in these transactions are discussed in the article. PMID- 9395963 TI - Merging HMOs into an all-payer system: a model to pursue? AB - An almost unending debate over the future shape of America's health care system increasingly focuses on the continuum between managed competition and government regulations, and the question: What is right for the United States? The Maryland all-payer, rate setting system for its 50 hospitals, where over a third of the state's population is enrolled in managed care plans, is used as an example of relatively successful blending of competitive and regulatory strategies. This article concludes with the theme that given America's penchant for compromise, competitive and regulatory approaches can coexist in a pluralistic health system that constrains the use of services and costs, and enhances access and the quality of patient care. PMID- 9395964 TI - HMO quality and financial performance: is there a connection? AB - Following the continuing expansion of managed care is a growing public interest in the quality of care provided by managed care organizations. Public and private organizations are actively developing systems for measuring and monitoring quality of care with the intention of holding plans more accountable for quality improvements and encouraging health care consumers to make responsible choices. This article examines correlations between a prevention-based quality measure and other commonly used measures of differentiating health plans, including financial performance and profit status. The findings are instructive in efforts to develop a quality measurement system that incorporates a variety of measures of plan performance. PMID- 9395965 TI - Unionism and professionalism--physician, do no harm: one person's view. PMID- 9395967 TI - Advanced practice nurses. PMID- 9395966 TI - Financing universal health care coverage for America's children. AB - The Clinton administration is about to engage in finding a solution to the much heralded bankruptcy of Medicare. Still, it is appropriate to consider that millions of this nation's children lack health insurance. A plan to provide such coverage through universal insurance would cover a set of defined benefits and be means tested at the upper end of incomes. It would create a rational replacement for a hodgepodge of entitlement programs based on actuarially sound principles. It would derive funds from current Medicaid allocations and private employer contributions for minor dependents. PMID- 9395968 TI - Sore nipples. PMID- 9395969 TI - Breastfeeding after early discharge. PMID- 9395970 TI - Patient self-testing. PMID- 9395971 TI - Electronic fetal heart rate monitoring: research guidelines for interpretation. The National Institute of Child Health and Human Development Research Planning Workshop. AB - The purpose of the National Institutes of Health (NIH) research planning workshops are to assess the research status of clinically important areas. This article reports on a workshop, whose meetings were held between May 1995 and November 1996, in Bethesda, MD, and Chicago, IL. Its specific purpose was to develop standardized and unambiguous definitions for fetal heart rate (FHR) tracings. Their recommendations for interpreting FHR patterns are being published here, in JOGNN, and simultaneously by the American Journal of Obstetrics and Gynecology. PMID- 9395972 TI - What nurses should know about natural family planning. AB - Two common natural family planning (NFP) methods are the ovulation method based on characteristics of cervical mucus and the symptothermal method based on changes in cervical mucus, basal body temperature, and the cervix. Both methods are effective when used correctly. Nurses should understand the principles of NFP and introduce these methods in discussions of family planning options. Interested clients should be referred to a certified NFP instructor for education and supervision. PMID- 9395973 TI - Addressing couples' sexuality concerns during the childbearing period: use of the PLISSIT model. AB - During pregnancy, a couple may benefit from discussing sexuality concerns with a nurse. Couples indicate they do not receive this support, and frequently nurses state they do not have the knowledge, time, or skills to provide patient education regarding sexuality. The PLISSIT model provides a framework for developing and implementing interventions to assist clients in maintaining their sexual relationship throughout the childbearing experience. PMID- 9395974 TI - Development of a pacifier for low-birth-weight infants' nonnutritive sucking. AB - Nonnutritive sucking can provide low-birth-weight infants with an opportunity to organize their behavior, an important component of developmental care. A pacifier specifically designed for low-birth-weight infants facilitates their nonnutritive sucking to more fully meet their needs. The research and development of a pacifier for low-birth-weight infants incorporated a naturalistic approach and used the best model, the infant thumb, in the design. Clinical trials with infants randomized to control and experimental groups were conducted to compare the prototype pacifier to a commercially available pacifier. Observations using the Anderson Behavioral State Scale demonstrated that infants using the prototype pacifier more often were found to be in an alert state. This pacifier may contribute to infants' state organization for optimum feeding and could be a component in developmental care planning. PMID- 9395975 TI - Caring for childbearing Korean women. AB - This article examines the traditional and modern cultural elements that may influence the health behaviors of the childbearing Korean woman and suggests ways to provide culturally sensitive care. The first author, born and raised in Korea, shares her reflections of culture and examples of clinical situations in Korea. Implications for nursing care are addressed through specific cultural prescriptions. Do's and don'ts are presented to foster culturally appropriate care for Korean childbearing women. PMID- 9395976 TI - The nurse-technology relationship: the case of ultrasonography. AB - OBJECTIVE: To examine the nurse-technology relationship via a case study of ultrasonography in women's health nursing practice. DESIGN: Exploratory case study using ethnographic methods. SETTING: Outpatient obstetric and gynecology clinic of a large teaching hospital. PARTICIPANTS: Three nurses, two technicians, and one physician. RESULTS: Two of the nurses and the physician were very "nurse like" in their use of the technology of ultrasonography to fulfill the traditional nursing purposes of observation, teaching, and comforting. The third nurse was nurse-like in talk, but at times "technician-like" in performance, allowing the technology to pull her away from nursing purposes. CONCLUSIONS: The expert nurses bent the technology to nursing purposes. Sonographers performed in accordance with professional training and experience, rather than with gender expectations. The subtle differences between expert nurse use and technical use (by novice nurses or technicians) of ultrasonography may provide a strong rationale against current efforts to substitute technically trained or technically oriented personnel for expert nurses. PMID- 9395977 TI - Social support, knowledge of infant development, and maternal confidence among adolescent and adult mothers. AB - OBJECTIVE: To explore the influence of social support and knowledge of infant development on maternal confidence in performing infant care tasks among adolescents and adults. DESIGN: Descriptive, exploratory. SETTING: A large urban teaching hospital in the midwestern United States. PARTICIPANTS: 116 adolescent mothers (ages 13 to 19 years); 101 adult mothers (ages 20 to 41 years). MAIN OUTCOME MEASURES: Adult mothers reported higher levels of knowledge about infant development than did adolescent mothers. For both adolescent and adult mothers, social support and knowledge of infant development were significantly correlated with confidence in providing infant care. For adolescents, knowledge of infant development explained 15% of the variance in maternal confidence scores, and the number of people in the adolescent's household explained an additional 5%. For adults, parity explained 10%, social support explained 9%, and knowledge of infant development explained 4% of the variance in maternal confidence scores. CONCLUSIONS: An adolescent's knowledge of infant development significantly affects her confidence in providing infant care. Given the learning needs of new mothers and the time constraints for in-hospital postpartum teaching, nurses may need to adjust content and teaching methods for all new mothers, but especially to meet the needs of adolescent mothers. PMID- 9395978 TI - In utero exposure to terbutaline: effects on infant behavior and maternal self esteem. AB - OBJECTIVE: Little is known about correlates of infant behavior after tocolytic intervention. This study investigated three related questions: (a) Are there any behavioral differences in infants exposed in utero to terbutaline and infants not exposed? (b) Is the mother's perception of her infant influenced by group status? (c) Is maternal self-esteem influenced by group status? DESIGN: Group comparison over time of infants exposed in utero to terbutaline and infants not exposed. Group mean cluster scores and recovery curves were compared. Mothers' responses to a series of questionnaires were compared according to group status. The first author remained blind to group status until all measures were completed on all participants. SETTING: Recruitment and first administration of the Brazelton Neonatal Behavioral Assessment Scale occurred in two regional hospitals, at Minneapolis and St. Paul. Subsequent administrations took place in the infants' homes. PARTICIPANTS: Forty-eight healthy, full-term (> or = 37 weeks gestational age) neonates and their mothers, 16 of whom had been given terbutaline, and 32 control subjects. MAIN OUTCOME MEASURES: Brazelton Neonatal Behavioral Assessment Scale was administered three times to each neonate. Mothers completed the Infant Characteristics Questionnaires, the Neonatal Perception Inventories, and the Maternal Self-Report Inventory. RESULTS: Significant differences were found between the two groups of infants on the habituation cluster at Time 1 and the autonomic stability cluster at Time 1 and Time 2. No significant group differences were found in the mothers' responses to the questionnaires. CONCLUSIONS: Nurses can assure parents that neonates exposed to terbutaline, although they may exhibit initial difficulties with behavioral organization, recover well over the course of the neonatal period. In addition, nurses can help mothers discover methods that will help calm their newborns. PMID- 9395979 TI - Strategies for designing a research utilization project with labor and delivery nurses. AB - The background and development of the second National AWHONN Research Utilization Project on Second Stage Labor Management that was conducted in multiple sites within the United States and Canada are presented. On the basis of the results of the project, recommendations for designing other research utilization projects are discussed. PMID- 9395980 TI - The process for initiating nursing practice changes in the intrapartum: findings from a multisite research utilization project. AB - The process of implementing a research-based protocol (the Second Stage Labor Nursing Management) at 40 sites in North America is described. Both positive and negative factors involved in implementing and adhering to the protocol are presented based on the reports of site coordinators. Key findings from the process data are: (a) the term "research utilization" causes confusion, (b) it is essential that nurses collaborate with other disciplines when attempting to change practice, (c) administrative endorsement of research utilization is important for practice change to occur, (d) nurses know their own practice sites and how to facilitate protocol acceptance, and (e) practice change may not need to occur all at once. PMID- 9395981 TI - Pushing techniques during labor: issues and controversies. AB - The lack of support for spontaneous bearing down versus directed pushing efforts, varying opinions on the determination of readiness for pushing, and the prevailing use of prolonged breath holding associated with pushing during labor are aspects of second-stage labor management that continue to be areas of contention among physicians and nurses. A discussion of current practice outcomes surrounding these controversies from the AWHONN Second Stage Labor Research Utilization Project conducted in 1994-1995 is presented in view of the available research literature. In addition, recommendations for future nursing research are identified. PMID- 9395982 TI - Positioning during the second stage of labor: moving back to basics. AB - The advantages of an upright position during labor are presented, with historic, physiologic, and psychosocial aspects discussed. The influences of modern obstetric practices such as electronic fetal monitoring and anesthesia practices are discussed with findings related to the use of upright positions from the Association of Women's Health, Obstetric, and Neonatal Nursing National Research Utilization Project on Second Stage Labor Management integrated. Recommendations for facilitating upright positions on the labor and delivery unit are presented. PMID- 9395983 TI - Safe closure of therapeutic relationships with abuse survivors. AB - 1 Safe closure of a therapeutic relationship is needed because neither healer nor client can be a "stranger" again. 2 A "failed termination" may overshadow the therapeutic gains that were made during the working phase, constituting a "retraumatization." 3 Closure, as opposed to termination, involves a unique, individualized mutual transformation of the therapeutic relationship into a "real relationship." PMID- 9395984 TI - Use of analogy to educate clients about the roles of neurotransmitters in addictions. AB - 1 The professional nurse who practices within the field of addictions is in an ideal position to serve as an educational conduit and catalyst for promoting understanding of the addictive process. 2 Historically, the use of analogy as a teaching device can be seen in the readings of the Bible to as far back as Plato. 3 The client's level of functioning must be assessed by the professional nurse so that any educational approach or intervention will be designed to meet the appropriate level of the client. PMID- 9395985 TI - Advanced practice psychiatric-mental health nursing in a community-based nurse managed primary care program. AB - In the evolving health care environment, advanced practice opportunities for psychiatric-mental health nurses may lie in managed care models of community oriented primary health care. As providers, care managers, health promoters and educators, and client advocates with family systems expertise, psychiatric-mental health advanced practice specialists are a necessary component of all primary health care. This article describes the development of a community-based primary care practice that integrates physical and mental health care on the same site. PMID- 9395986 TI - Benefits of a nurse-psychiatrist collaborative practice in an anxiety disorders unit. AB - 1 Only one out of four people receives the appropriate therapy for anxiety disorders, despite available and effective treatments. 2 Nurse-doctors collaborative practice is a working relationship with a sharing of responsibilities of the treatment program within the realm of respective roles. 3 Using an evidence-based care approach allowed the care providers to continually monitor, assess and revise the treatment program which proved to be cost effective and time efficient, yet provided premium patient care. PMID- 9395987 TI - Religion/spirituality and health among elderly African Americans and Hispanics. AB - 1 It is important to view elders in a multicultural sense and also understand that there may be great heterogeneity within cultural or ethnic groups. 2 Knowledge of the impact of religion and spiritual beliefs for ethnic groups can help health care professionals design interventions that are culture-specific to the beliefs of individuals. 3 The psychiatric nurse is in a unique position to encourage the patient to use healthy religious practices to deal with their illness, whether mental or physical. PMID- 9395988 TI - Relationship between tolerance/intolerance of ambiguity and perceived environmental uncertainty in hospitals. AB - 1 Many variables may contribute to a nurse's perceptions when working in a changing health care environment, especially in the field of psychiatry. 2 While this study did not find a relationship between tolerance for ambiguity and perceived environmental uncertainty in hospitals, age and educational background appear to be variable, which may warrant further study as ultimately impacting nursing. 3 Research which explores those personality variables which underlie perception would assist nurse administrators in designing interventions, opening new lines of communication, and increasing sensitivity to individual nurse's need in these changing times. PMID- 9395989 TI - The aesthetic aspects of psychoanalysis. PMID- 9395990 TI - In Electra's voice: Gilligan's debt to Freud. PMID- 9395991 TI - Malignant eroticized countertransference. AB - Gabbard (1994) divided the pathology of therapists, both male and female, who commit sexual boundary violations into those who are psychotic, those who are predatory psychopaths, those engaging in masochistic surrender, and those called "the lovesick therapist." Lovesick therapists are the most common type and manifest crucial narcissistic themes of "a desperate need for validation by their patients, a hunger to be loved and idealized, and a tendency to use patients to regulate their own self-esteem" (p. 127). Among the psychodynamic aspects of this curiously circumscribed area of loss of reality testing that makes it difficult for the therapist to see how self-destructive and harmful such enactment is, are an unconscious reenactment of incestuous longings, a misperception of the patient's wish for maternal nurturance as a sexual overture, enactments of rescue fantasies, a projected idealization of the self of the therapist, a confusion of the therapist's needs with the patient's needs, a fantasy that love is curative, acting out disavowed rage at the patient, or rage at an organization, an institute, or one's training analyst, a manic defense against mourning, a narcissistic fantasy that their sexual affair is an exception, insecurity regarding masculine identity, and assorted primitive preoedipal themes. Gabbard's (1991) erotized countertransference is one variety of what I have termed malignant eroticized countertransference. His variety is a development that occurs under the pressure of the patient's preemptive and compelling expressions of lust and love, the patient's erotic transference. But malignant eroticized countertransference can also occur without the patient having offered any such expressions; it can even occur on first meeting the patient when he or she walks into the office! This is akin to the romantic "love-at-first-sight" theme so favored in the movies and by novelists, but it is always pathological when it occurs in the therapeutic situation. Countertransference enactments are a creation between the patient and the therapist on a continuum from one pole, where the patient has just walked into the office and contributes almost nothing directly, to the other pole, where the therapist loses control of himself or herself as a response to the unbearable pressure of the patient's lust. In the treatment of malignant eroticized countertransference it seems clear from this discussion that every case should be evaluated psychodynamically and the treatment should be made to fit the patient, not the patient to the treatment. Each situation should be studied in psychodynamic depth without preconceptions based on generalizations or formulas. Therapists who are psychotic should of course be treated with antipsychotic drugs and usually should not be allowed to practice any further. Therapists who are psychopathic or sociopathic predators should certainly never be allowed to practice. Some of the individuals who are "lovesick," or, as I put it "love/lust obsessed," or those who have made a masochistic surrender to a sadistic destructive patient, are in need of reanalysis and have the potential to continue as effective therapists under careful supervision. Therapists like this do not deserve to be summarily dismissed from the profession but, like therapists who develop other serious neurotic problems, should receive appropriate help from us. PMID- 9395992 TI - Institutional countertransference: the matrix of social structure and psychic structure. PMID- 9395993 TI - Moral values: development and gender influences. PMID- 9395994 TI - Time compressed: psychoanalysis in the days of HIV and AIDS. PMID- 9395995 TI - The hospital ethics committee: a role for the contemporary psychoanalyst. PMID- 9395996 TI - A model for dealing with countertransference in first-year psychiatric residents. PMID- 9395997 TI - Interminable analysis? PMID- 9395998 TI - Further reflections on creativity and personal growth: sculpture and poetry. PMID- 9395999 TI - Preservation of pig liver allografts after warm ischemia: normothermic perfusion versus cold storage. AB - Warm ischemia is known to induce substantial damage to the liver parenchyma. With respect to clinical liver transplantation, the tolerance of the liver to warm ischemia and the preservation of these organs have not been studied in detail. In isolated reperfused pig livers we proceeded according to the following concept: Livers were subjected to 1 or 3 h of warm ischemia. Subsequently, these organs were preserved by either normothermic perfusion or cold storage (histidine tryptophan-alpha-ketoglutarate, HTK) for 3 h each. After storage, liver function was assessed in a reperfusion circuit for another 3 h. Parameters under evaluation were bile flow, perfusion flow, oxygen consumption, enzyme release into the perfusate (creatine kinase, glutamic oxaloacetic transaminase (GOT), lactic dehydrogenase, and glutamic pyruvic transaminase), and histomorphology. Damage to the liver was lowest after warm ischemia of 1 h. The results after cold storage were superior to those after normothermic perfusion (GOT: 3.2 +/- 0.3 and 2.6 +/- 0.2 U/g liver; cumulative bile production: 14.7 +/- 2.1 and 9.4 +/- 1 ml, respectively; P < 0.05). In contrast, we found substantial damage at the end of reperfusion in livers undergoing 3 h of warm ischemia under both preservation techniques with severe hepatocellular pyknoses and essentially altered nonparenchymal cells. The results suggest that pig livers undergoing 1 h of warm ischemia and cold storage for 3 h with HTK solution may lead to functioning after transplantation. PMID- 9396000 TI - Survival in primary soft tissue sarcoma of the extremities and trunk. AB - BACKGROUND: Soft tissue sarcomas (STS) of the extremities are rare. The purpose of this study was to identify prognostic risk factors associated with survival in patients with primary extremity and truncal STS. METHODS: Patient, tumor, and pathologic data from 149 consecutive patients with localized primary STS of the extremities and trunk were analyzed using Kaplan-Meier and Cox regression techniques to identify univariate and multivariate risk factors. A subgroup analysis was performed to compare factors predictive of survival in patients who received treatment before (n = 50) and after (n = 99) treatment was standardized in 1988. RESULTS: The 5-year survival rate was 76.5% with an average follow-up of 6 years. Local recurrence occurred in 23% of all patients, 40% before 1988 and 15% after 1988 (P < 0.0001). Risk factors associated with survival included resection quality (R0 vs. R1; P < 0.0001), era of operation (P = 0.002), local recurrence (P < 0.001), UICC stage (P < 0.0001), tumor size (P < 0.001), tumor depth (P = 0.002), regional lymph nodes (P < 0.0001), and histology (P < 0.0001). Multivariate analysis revealed that tumor size, tumor depth, and resection quality were independent risk factors of survival. CONCLUSIONS: These results indicate that management of STS in a specialized institution improves overall survival. Resection quality is the most important risk factor of survival. Therefore, effort should be made during primary treatment of STS to achieve wide, tumor-free resection margins. PMID- 9396001 TI - Quantitative measurements of released amines from individual exocytosis events. AB - Chemical analysis of single cells is an area of great interest in the biological sciences. Single-cell systems are being utilized as a model to understand in vivo processes better. One method that is moving to the forefront in cellular analysis is electrochemistry. Owing to their rapid response time and small dimensions, voltammetric microelectrode techniques, such as amperometry and fast-scan voltammetry, have made it possible to monitor minute amounts of biological compounds and transiently occurring chemical events in cellular systems. The application of these methods to the quantitation of individual vesicular release events from single cells is overviewed here. The application of electrochemical monitoring to several types of cultured cells, including bovine adrenal chromaffin cells, rat pheochromocytoma (PC12) cells, beige mouse mast cells, superior cervical ganglion neurons, and human pancreatic beta-cells, as well as to the invertebrate systems, the leech Hirudo medicinalis, and pond snail Planorbis corneus has provided a wealth of new information concerning exocytosis. Results obtained from the studies highlight the potential of electrochemical techniques in cellular analysis to contribute to our understanding of molecular and pharmacological effects on exocytosis. This article overviews work done on all the above cell types with an emphasis on PC12 cells. PMID- 9396002 TI - Acetylcholine and associative memory in the piriform cortex. AB - The significance of cholinergic modulation for associative memory performance in the piriform cortex was examined in a study combining cellular neurophysiology in brain slices with realistic biophysical network simulations. Three different physiological effects of acetylcholine were identified at the single-cell level: suppression of neuronal adaptation, suppression of synaptic transmission in the intrinsic fibers layer, and activity-dependent increase in synaptic strength. Biophysical simulations show how these three effects are joined together to enhance learning and recall performance of the cortical network. Furthermore, our data suggest that activity-dependent synaptic decay during learning is a crucial factor in determining learning capability of the cortical network. Accordingly, it is predicted that acetylcholine should also enhance long-term depression in the piriform cortex. PMID- 9396003 TI - Caffeine and the olfactory bulb. AB - Caffeine, a popular CNS stimulant, is the most widely used neuroactive drug. Present in coffee, tea, chocolate, and soft drinks as well as over-the-counter and prescription medications, it influences millions of users. This agent has achieved recent notoriety because its dependency consequences and addictive potential have been re-examined and emphasized. Caffeine's central actions are thought to be mediated through adenosine (A) receptors and monoamine neurotransmitters. The present article suggests that the olfactory bulb (OB) may be an important site in the brain that is responsible for caffeine's central actions in several species. This conclusion is based on the extraordinarily robust and selective effects of caffeine on norepinephrine (NE), dopamine (DA), and particularly serotonin (5HT) utilization in the OB of mice. We believe that these phenomena should be given appropriate consideration as a basis for caffeine's central actions, even in primates. Concurrently, we review a rich rodent literature concerned with A, 5HT, NE, and DA receptors in the OB and related structures along with other monoamine parameters. We also review a more limited literature concerned with the primate OB. Finally, we cite the literature that treats the dependency and addictive effects of caffeine in humans, and relate the findings to possible olfactory mechanisms. PMID- 9396004 TI - Current issues in invertebrate phototransduction. Second messengers and ion conductances. AB - Investigation of phototransduction in invertebrate photoreceptors has revealed many physiological and biochemical features of fundamental biological importance. Nonetheless, no complete picture of phototransduction has yet emerged. In most known cases, invertebrate phototransduction involves polyphosphoinositide and cyclic GMP (cGMP) intracellular biochemical signaling pathways leading to opening of plasma membrane ion channels. Excitation is Ca(2+)-dependent, as are adaptive feedback processes that regulate sensitivity to light. Transduction takes place in specialized subcellular regions, rich in microvilli and closely apposed to submicrovillar membrane systems. Thus, excitation is a highly localized process. This article focuses on the intracellular biochemical signaling pathways and the ion channels involved in invertebrate phototransduction. The coupling of signaling cascades with channel activation is not understood for any invertebrate species. Although photoreceptors have features that are common to most or all known invertebrate species, each species exhibits unique characteristics. Comparative electrophysiological, biochemical, morphological, and molecular biological approaches to studying phototransduction in these species lead to fundamental insights into cellular signaling. Several current controversies and proposed phototransduction models are evaluated. PMID- 9396005 TI - Alcohol, astroglia, and brain development. AB - Glial cells constitute one of the most common cell types in the brain. They play critical roles in central nervous system (CNS) development. Recent evidence demonstrates that glial cells are profoundly affected by prenatal alcohol exposure, suggesting that alterations in these cells may participate in CNS abnormalities associated with ethanol-induced teratogenesis. In vivo studies show that prenatal exposure to alcohol hampers myelinogenesis and is associated with neuroglial heterotopias and abnormal astrogliogenesis. Studies using primary cultures of rat cortical astrocytes show that ethanol affects DNA, RNA, and protein synthesis, decreases the number of mitotic cells, alters the content and distribution of several cytoskeletal proteins including the astroglial marker, glial fibrillary acidic protein (GFAP), and the levels of plasma-membrane glycoproteins, reduces the capacity of astrocytes to secrete growth factors, and induces oxidative stress. Furthermore, ethanol exposure during early embryogenesis alters the normal development of radial glia cells (the main astrocytic precursors), delays the onset of GFAP expression, and decreases mRNA GFAP levels in fetal and postnatal brains and in radial glia and astrocytes in primary culture. Recent evidence suggests that ethanol interferes with the transcription process of GFAP, thus leading to a reduction in GFAP-gene expression during astrogliogenesis. However, brief exposure of rats to high levels of ethanol during the neonatal period (the period of astrocyte differentiation) causes a transient gliosis, with an increase in GFAP and its mRNA levels. These findings indicate that astroglial cells are an important target of ethanol toxicity during central nervous system (CNS) development. PMID- 9396006 TI - Signals that initiate myelination in the developing mammalian nervous system. AB - The myelination of axons by oligodendrocytes in the central nervous system and Schwann cells in the peripheral nervous system is essential for the establishment of saltatory conduction. In the absence or destruction of the myelin sheath, as seen in demyelinating diseases, impulse conduction is impeded resulting in severe sensory and motor deficits. Axon myelination is the culmination of a sequence of events that begins with the differentiation of glial cells and proceeds to the transcription and translation of myelin genes, the elaboration of a myelin sheath, and the recognition and ensheathment of axons. This review examines the regulatory mechanisms for each of these steps and compares and contrasts the role of the axon in initiating myelination in the central and peripheral nervous system. PMID- 9396007 TI - Nucleotide receptors in the nervous system. An abundant component using diverse transduction mechanisms. AB - Extracellular nucleotides achieve their role as cell-to-cell communicators by acting at cell surface transmembrane receptors-the P2 receptors. Before molecular cloning led to the isolation of any P2-receptor sequence, a small number of receptor types had been proposed on the basis of pharmacological evidence. The application of molecular biology to this field of receptor research has indicated that a great underestimation of the number of receptor subtypes and of their abundance had occurred. There are now known to be seven characterized P2Y (G protein linked) receptors and the same number again of P2X receptors of the transmitter-gated ion channel type. In this review, we discuss the properties of these cloned receptors, their distribution within the nervous system, and their methods of signal transduction. PMID- 9396008 TI - RC3/neurogranin, a postsynaptic calpacitin for setting the response threshold to calcium influxes. AB - In this review, we attempt to cover the descriptive, biochemical and molecular biological work that has contributed to our current knowledge about RC3/neurogranin function and its role in dendritic spine development, long-term potentiation, long-term depression, learning, and memory. Based on the data reviewed here, we propose that RC3, GAP-43, and the small cerebellum-enriched peptide, PEP-19, belong to a protein family that we have named the calpacitins. Membership in this family is based on sequence homology and, we believe, a common biochemical function. We propose a model wherein RC3 and GAP-43 regulate calmodulin availability in dendritic spines and axons, respectively, and calmodulin regulates their ability to amplify the mobilization of Ca2+ in response to metabotropic glutamate receptor stimulation. PEP-19 may serve a similar function in the cerebellum, although biochemical characterization of this molecule has lagged behind that of RC3 and GAP-43. We suggest that these molecules release CaM rapidly in response to large influxes of Ca2+ and slowly in response to small increases. This nonlinear response is analogous to the behavior of a capacitor, hence the name calpacitin. Since CaM regulates the ability of RC3 to amplify the effects of metabotropic glutamate receptor agonists, this activity must, necessarily, exhibit nonlinear kinetics as well. The capacitance of the system is regulated by phosphorylation by protein kinase C, which abrogates interactions between calmodulin and RC3 or GAP-43. We further propose that the ratio of phosphorylated to unphosphorylated RC3 determines the sliding LTP/LTD threshold in concept with Ca2+/ calmodulin-dependent kinase II. Finally, we suggest that the close association between RC3 and a subset of mitochondria serves to couple energy production with the synthetic events that accompany dendritic spine development and remodeling. PMID- 9396013 TI - Decreased nocturnal secretion of melatonin in drug-free schizophrenics: no change after subchronic treatment with antipsychotics. AB - To evaluate the biosynthetic activity of the pineal gland in schizophrenia, the circadian rhythm of plasma melatonin was investigated in 9 drug-free chronic schizophrenics and in matched healthy subjects. In 7 of the patients, the 24-hour secretory pattern of the pineal hormone was reassessed after 10 weeks of treatment with antipsychotic drugs. In drug-free schizophrenics, the nocturnal increase in plasma melatonin levels was significantly blunted as compared to healthy subjects (p < 0.0001). Chronic treatment with antipsychotic drugs significantly improved psychotic symptomatology, but did not change the secretory pattern of melatonin. These data show that the biosynthetic activity of the pineal gland is impaired in chronic schizophrenia and that successful treatment with antipsychotic drugs does not go parallel with changes in the production of melatonin. PMID- 9396014 TI - Weak association between blood sodium, potassium, and calcium and intensity of symptoms in major depressed patients. AB - In previous reports, we showed that plasma and erythrocyte magnesium were increased in many drug-free hospitalized depressed patients. Furthermore, we observed that erythrocyte magnesium content was related to the intensity of the symptoms. Highly depressed patients had the highest magnesium values. Today, we report the results of plasma and erythrocyte sodium and potassium, and of total and ultrafilterable plasma calcium in 66 hospitalized patients with major depression compared to 58 healthy controls. No consistent differences in these biochemical parameters are observed between patients when separated according to intensity of anxiety, psychomotor retardation, and moral distress. Plasma sodium is higher and plasma potassium lower in female patients of all subgroups as compared to controls. Both male patients and controls have erythrocyte sodium and potassium levels that are significantly different from those of females. This clearly suggests a separation into genders in such studies. In conclusion--in contrast to blood magnesium--sodium, potassium, and calcium levels do not seem to be related to the intensity of the main clinical symptoms in hospitalized patients with major depression. PMID- 9396012 TI - Gene therapy for Parkinson's disease. AB - Gene therapy is a potentially powerful approach to the treatment of neurological diseases. The discovery of neurotrophic factors inhibiting neurodegenerative processes and neurotransmitter-synthesizing enzymes provides the basis for current gene therapy strategies for Parkinson's disease. Genes can be transferred by viral or nonviral vectors. Of the various possible vectors, recombinant retroviruses are the most efficient for genetic modification of cells in vitro that can thereafter be used for transplantation (ex vivo gene therapy approach). Recently, in vivo gene transfer to the brain has been developed using adenovirus vectors. One of the advantages of recombinant adenovirus is that it can transduce both quiescent and actively dividing cells, thereby allowing both direct in vivo gene transfer and ex vivo gene transfer to neural cells. Probably because the brain is partially protected from the immune system, the expression of adenoviral vectors persists for several months with little inflammation. Novel therapeutic tools, such as vectors for gene therapy have to be evaluated in terms of efficacy and safety for future clinical trials. These vectors still need to be improved to allow long-term and possibly regulatable expression of the transgene. PMID- 9396009 TI - Vesicular neurotransmitter transporters. Potential sites for the regulation of synaptic function. AB - Neurotransmission depends on the regulated release of chemical transmitter molecules. This requires the packaging of these substances into the specialized secretory vesicles of neurons and neuroendocrine cells, a process mediated by specific vesicular transporters. The family of genes encoding the vesicular transporters for biogenic amines and acetylcholine have recently been cloned. Direct comparison of their transport characteristics and pharmacology provides information about vesicular transport bioenergetics, substrate feature recognition by each transporter, and the role of vesicular amine storage in the mechanism of action of psychopharmacologic and neurotoxic agents. Regulation of vesicular transport activity may affect levels of neurotransmitter available for neurosecretion and be an important site for the regulation of synaptic function. Gene knockout studies have determined vesicular transport function is critical for survival and have enabled further evaluation of the role of vesicular neurotransmitter transporters in behavior and neurotoxicity. Molecular analysis is beginning to reveal the sites involved in vesicular transporter function and the sites that determine substrate specificity. In addition, the molecular basis for the selective targeting of these transporters to specific vesicle populations and the biogenesis of monoaminergic and cholinergic synaptic vesicles are areas of research that are currently being explored. This information provides new insights into the pharmacology and physiology of biogenic amine and acetylcholine vesicular storage in cardiovascular, endocrine, and central nervous system function and has important implications for neurodegenerative disease. PMID- 9396010 TI - Nicotinic acetylcholine receptors in health and disease. AB - Nicotinic acetylcholine receptors (AChRs) are a family of acetylcholine-gated cation channels that form the predominant excitatory neurotransmitter receptors on muscles and nerves in the peripheral nervous system. AChRs are also expressed on neurons in lower amounts throughout the central nervous system. AChRs are even being reported on unexpected cell types such as keratinocytes. Structures of these AChRs are being determined with increasing precision, but functions of some orphan subunits are just beginning to be established. Functional roles for postsynaptic AChRs in muscle are well known, but in neurons the post-, peri-, extra-, and presynaptic roles of AChRs are just being revealed. Pathogenic roles of AChRs are being discovered in many diseases involving mechanisms ranging from mutations, to autoimmune responses, to the unknown; involving cell types ranging from muscles, to neurons, to keratinocytes; and involving signs and symptoms ranging from muscle weakness to epilepsy, to neurodegenerative disease, to psychiatric disease, to nicotine addiction. Awareness of AChR involvement in some of these diseases has provoked new interests in development of therapeutic agonists for specific AChR subtypes and the use of expressed cloned AChR subunits as possible immunotherapeutic agents. Highlights of recent developments in these areas will be briefly reviewed. PMID- 9396011 TI - Neural roles of immunophilins and their ligands. AB - The immunophilins are a family of proteins that are receptors for immunosuppressant drugs, such as cyclosporin A, FK506, and rapamycin. They occur in two classes, the FK506-binding proteins (FKBPs), which bind FK506 and rapamycin, and the cyclophilins, which bind cyclosporin A. Immunosuppressant actions of cyclosporin A and FK506 derive from the drug-immunophilin complex binding to and inhibiting the phosphatase calcineurin. Rapamycin binds to FKBP and the complex binds to Rapamycin And FKBP-12 Target (RAFT). RAFT affects protein translation by phosphorylating p70-S6 kinase, which phosphorylates the ribosomal S6 protein, and 4E-BP1, a repressor of protein translation initiation. Immunophilin levels are much higher in the brain than in immune tissues, and levels of FKBP12 increase in regenerating neurons in parallel with GAP-43. Immunophilin ligands, including nonimmunosuppressants that do not inhibit calcineurin, stimulate regrowth of damaged peripheral and central neurons, including dopamine, serotonin, and cholinergic neurons in intact animals. FKPB12 is physiologically associated with the ryanodine and inositol 1,4,5-trisphosphate (IP3) receptors and regulates their calcium flux. By influencing phosphorylation of neuronal nitric oxide synthase, FKBP12 regulates nitric oxide formation, which is reduced by FK506. PMID- 9396016 TI - Differential development of the enhanced metabolic response during amphetamine sensitization. AB - Behavioral sensitization elicited by repeated administration of amphetamine does not fully develop until a period after discontinuation of amphetamine, but then persists undiminished for a long time. This experiment investigated the regional metabolic changes in rats pretreated with amphetamine and challenged after different abstinence periods (2, 7 and 28 days), using the 2-[14C]deoxyglucose method. The results demonstrated that chronic amphetamine administration enhanced rates of local cerebral glucose utilization in specific cerebral regions. The magnitude and distribution of effects varied with the abstinence period. A challenge dose of d-amphetamine 2 days after pretreatment was found to have no more, or only mildly elevated, local cerebral glucose utilization compared with that following a single acute dose. In rats challenged at the 7th and 28th day, a supersensitive metabolic response was found in dopaminergic and non-dopaminergic areas. This finding suggested regional differences in the development of sensitization and underscored the importance of an abstinence period in the study of sensitization and amphetamine psychosis. PMID- 9396015 TI - Side effect profile of azathioprine in the treatment of chronic schizophrenic patients. AB - Various findings suggest auto-immune changes in schizophrenia. We have recently demonstrated that platelets from schizophrenic patients bear autoantibodies (PAA) which cross-react with brain antigens. Accordingly, treatment of schizophrenia with an immunosuppressant might be of potential benefit. In a recent case study, a chronic schizophrenic patient treated with azathioprine has demonstrated a clear psychiatric improvement preceded by a decrease in PAA level. A phase I study designed for assessing side-effects of short-term azathioprine treatment in a group of schizophrenic patients is described here. From a group of 40 chronic non-responsive patients, 14 patients demonstrating high PAA level have entered the study and 11 have complied all along. Two groups were tested in parallel. In the first (6 patients) 150 mg/day was given for 7 weeks while in the second (5 patients) the same regimen was given for two periods of 7 weeks with an interval of 6 weeks. Blood biochemistry and cell count, as well as determination of PAA were carried out weekly, starting 3 weeks before the trial and continuing up to 7 weeks after the treatment. Two out of 11 patients developed leucopenia in week 4. No other side-effects were recorded in any of the patients. A substantial reduction in PAA was observed in 3 out of 6 patients in group I and 4 out of 5 in group II. Two patients showed improvement of psychiatric symptomatology. Our results demonstrate that short-term azathioprine treatment induces transient leucopenia in 18% of the patients receiving the drug, much alike the percentage reported for other patient populations. PMID- 9396018 TI - The effects of 6R-L-erythro-5,6,7,8-tetrahydrobiopterin on ethanol-induced sleep time and ethanol elimination in inbred strains of mice with different alcohol preference. AB - 6R-L-erythro-5,6,7,8-tetrahydrobiopterin (6R-BH4) is a coenzyme for tyrosine, tryptophan and phenylalanine hydroxylases. Male C57BL/6J and DBA/2J mice were given 6R-BH4 (0, 12.5, 25.0 and 50.0 mg/kg of body weight, i.v.) or saline, and then 4 h later, all animals were injected with ethanol (EtOH) (4.0 g/kg, i.p.), which causes them to lose the righting reflex, to investigate the differences in EtOH-induced sleeping time. 6R-BH4 pretreatment reduced EtOH-induced sleep time in DBA/2J mice with lower alcohol preference (p < 0.05), which showed no changes in the pharmacokinetics of blood EtOH. No changes in EtOH-induced sleep time were observed in C57BL/6J mice with higher alcohol preference. These results indicate that the sensitivity to EtOH in mice with lower alcohol preference was associated with 6R-BH4 activity in the CNS. PMID- 9396017 TI - Exercise avoidance and impaired endurance capacity in patients with panic disorder. AB - Exercise habits and indices of aerobic fitness as measured by spiroergometric testing were examined in 38 patients with panic disorder and/or agoraphobia and 24 untrained healthy controls. Maximal oxygen consumption, maximal power output and the power output at a lactate concentration of 4 mmol/l were significantly reduced in the patient group when compared to untrained controls. Other parameters like physical work capacity at a heart rate of 150/min, maximal lactate concentration, vital capacity, subjective exertion at maximal work load, and maximal heart rate did not differ between patients and controls. Patient interviews revealed that aerobic exercise is avoided by the vast majority of patients. Reduced aerobic fitness might contribute to the pathophysiology of panic disorder and/or agoraphobia. PMID- 9396019 TI - On the use of neural network techniques to analyse sleep EEG data. First communication: application of evolutionary and genetic algorithms to reduce the feature space and to develop classification rules. AB - To automate sleep stage scoring, the system sleep analysis system to challenge innovative artificial networks (SASCIA) has been developed and implemented. The aims of our investigation were twofold: In addition to automatic sleep stage scoring the hypothesis was tested that the information of only 1 EEG channel (C4 A2) should be sufficient to automatically generate sleep profiles which are comparable with profiles made by sleep experts on the basis of at least 3-channel EEG (C4-A2), EOG and EMG, as EOG and EMG are seen as epiphenomena during sleep and the full information about the sleep stage should--according to our hypothesis--be available in the EEG. The main components of the SASCIA sleep analysis system are designed to meet the requirements of flexible adaptation to the interindividual differences of the sleep EEG. The core of the SASCIA sleep analysis system consists of neural networks. Supervised learning was implemented and the experts' scorings were included into the learning set and test set. The feature selections out of a large number (118) are performed by genetic algorithms and the topologies of the networks are optimized by evolutionary algorithms. Different mathematical procedures were used to evaluate and optimize the efficiency of the system. The profiles generated by SASCIA are in reasonable agreement with the sleep stages scored by experts according to RKR. The development of the system is communicated in three parts: the first communication deals with the application of the neural network techniques using evolutionary and genetic algorithms and with the selection of feature space. The second communication shows the training of these evolutionary optimized network techniques with multiple subjects and the application of context rules, while the third communication shows an improvement in the robustness by the simultaneous application of 9 different networks obtained from 9 subject types which were used in combination with context rules. PMID- 9396020 TI - Melatonin effects in a patient with severe REM sleep behavior disorder: case report and theoretical considerations. AB - REM sleep behavior disorder (RBD) is so far a possibly underestimated yet well described sleep disorder. Its major impact is the vigorous sleep behavior that often results in injuries to the patient himself or to people sleeping nearby. We treated a 64-year-old male with a clinically and polysomnographically confirmed diagnosis of RBD with 3 mg melatonin, which led to a significant reduction of motor activity during sleep, as measured by actigraphy (p < 0.0001 in all analyzed movement parameters), and a full clinical recovery over a 5-month treatment period. RBD phenomena gradually returned after melatonin administration was stopped. After 2 months' treatment, polysomnography showed no major changes except an increase of REM sleep (13 vs. 17% of sleep period time) and a better preservation of REM-sleep-associated muscle atonia. Our results suggest that melatonin might be able to reinforce REM sleep in RBD patients by enhancing its active inhibition of motor activity. PMID- 9396021 TI - [New aspects of the mechanism of Ca2+ entry in non-excitable cells]. AB - The capacitative Ca2+ entry has been accepted to play a crucial role in Ca2+ signaling in non-excitable cells, and the mechanisms linking the depletion of intracellular Ca2+ stores to the activation of Ca2+ entry are presently the subject of intensive investigation. There are two hypotheses to explain the molecular mechanism of capacitative Ca2+ entry. One is that a diffusible second messenger released from intracellular Ca2+ stores may activate a Ca2+ channel at the plasma membrane. Numerous molecules, including CIF, cGMP, arachidonic acid, and a small G-protein, are proposed as the candidates for the diffusible messenger. In addition, the capacitative Ca2+ entry has been suggested to be modulated by protein phosphorylation/dephosphorylation. Another hypothesis is termed the "conformational coupling model". This model suggests that information between the IP3 receptor and a plasma membrane Ca2+ channel may be transferred directly through conformational protein-protein interactions. The plasma membrane Ca2+ channel involved in the capacitative Ca2+ entry is still neither purified nor cloned, but recent studies suggest that the mammalian homologue of the Drosophila protein Trp may function as a capacitative Ca2+ entry channel. PMID- 9396022 TI - [ATP receptors in the central nervous system]. AB - ATP receptors in the central nervous system (CNS) are divided into 2 major classes, ionotropic (P2Xn) and G protein-coupled (P2Yn) ATP receptors. P2Xn receptors, a member of the 2-transmembrane family, contain non-selective cation channels that may play a role in rapid synaptic transmission. Seven subtypes of P2Xn were reported so far. Although all of these subtypes are distributed in the CNS, P2X4 and P2X6 are most abundantly and widely distributed. P2X3 is distributed only in trigeminal ganglia neurons as well as in small-diameter DRG neurons, suggesting their relation to pain. P2Yn receptors, a member of the 7 transmembrane superfamily, are coupled with Gq/11 to activate PLC beta. These receptors are thought to play an important role in the modulation of synaptic efficacy. Seven subtypes of P2Yn were reported so far. P2Y1, P2Y2, P2Y3 and P2Y4 are distributed in the CNS. Neither selective agonists nor antagonists to P2Xn and P2Yn are known. PMID- 9396023 TI - [Pharmacological approach to control American foulbrood of honeybees]. AB - In this review, I will describe honeybee biology from my prospective as a veterinary pharmacologist and will provide a summary of my research project to search for effective drugs to control American foulbrood, a bacterial disease of honeybees. In conclusion, mirosamicin, a macrolide antibiotic, as a preventive and glutaral, an alkylating agent, as a disinfectant were the most promising drugs. PMID- 9396024 TI - [Measurement of receptor-operated Ca2+ influx by microspectrofluometry combined with the whole-cell patch clamp technique]. AB - Signal transduction from mouse bradykinin B2-receptors to Ca2+ influx was studied in single control or v-Ki-ras-transformed NIH/3T3 (DT) fibroblasts. Microspectrofluometry (fura-2) was combined with the whole-cell patch-clamp technique to study bradykinin-activated Ca2+ influx. Cytosolic Ca2+ oscillations observed at holding potentials of -20 to -80 mV were terminated by holding at -10 mV or more depolarized potentials. Bradykinin significantly enhanced the hyperpolarization-induced increases in the intracellular free Ca2+ concentration upon membrane hyperpolarization only in DT cells but not in control cells. Internal application of 10 microM inositol 1,3,4,5-tetrakisphosphate (InsP4) mimicked membrane potential-dependent Ca2+ entry. Activation of B2-receptors resulted in a decrease of cellular fluorescence at the excitation wavelength of 340 or 360 nm after MnCl2 application in DT cells. This Mn2+ entry through the Ca2+ influx pathway increased with membrane hyperpolarization below -20 mV. The results suggest that bradykinin-induced cytosolic Ca2+ oscillations in ras transformed NIH/3T3 cells are maintained by bradykinin-activated continuous Ca2+ influx, which may use Ins(1,3,4,5)P4 as an intracellular messenger. PMID- 9396025 TI - [Evaluation of marble burying behavior: induced alteration of monoamine metabolism in mouse brain]. AB - Evaluation of marble burying behavior (MBB) housing-induced alteration of monoamine metabolism in mouse brain was performed by measuring metabolite levels with HPLC-ECD. Isolated housing of mice, each in a cage (22 x 32 x 14 cm; sawdust, 1-mm diameter; 5 cm in thickness) with 15 evenly spaced glass marbles on the floor (2.5-cm diameter; control, without marbles) for 24-168 hr, 5-HIAA contents were decreased in three regions: the midbrain, thalamus and hypothalamus. 5-HT turnover was not inhibited except for in the hypothalamus due to the decreases of 5-HT in the other two regions. On the other hand, DOPAC content and DA turnover were decreased in four regions: striatum, midbrain, thalamus, hypothalamus. The decrease in hypothalamic monoamine neurons was observed notably after 72 hr of MBB housing. No alterations were observed in feeding, water-drinking, spontaneous locomotor activity, number of buried marbles, serum corticosterone and serum glucose concentrations during MBB keeping compared with the control mice. These results suggested that the isolated mouse housed in a cage with evenly spaced glass marbles for a long period may be a model animal for alteration of monoamine metabolism in brain regions without physical infringement. PMID- 9396026 TI - [Constructional apraxia in right brain-damaged patients]. PMID- 9396027 TI - [Clinical study on air in epidural hematomas]. AB - Air in epidural hematoma has previously been reported by some authors. With the advent of CT scan, the presence of air in epidural hematomas is not uncommon findings. In our series, 27 of 78 (34.6%) cases with acute epidural hematoma had air bubbles in epidural hematoma on CT scan. Acute epidural hematoma was located in the temporal in 18, frontal in 3, occipital in 2, parietal in one and posterior fossa in 3 cases. The air entrance was thought to be mastoid air cells in 13, open fracture in 5, frontal sinus in 3, sphenoid sinus in 3, and unknown in 3 cases. No patients in our series developed meningeal infection or tension pneumocephalus. There was no statistical difference of overall outcome or risk of increase in size of hematoma between acute epidural hematoma with and without air. PMID- 9396028 TI - [Electrophysiological evaluation of polyneuropathy in juvenile insulin dependent diabetics]. AB - Thirty patients with juvenile insulin dependent diabetes mellitus (IDDM) were electrophysiologically evaluated. In addition to the conventional motor and sensory nerve conduction studies, intrafascicular microneurography was performed in the median nerve. In this method a tungsten microelectrode was inserted into the median nerve trunk at the elbow, and a compound nerve action potential (CNAP) was recorded with supramaximal electrical stimulation at the wrist. The subjects' age ranged from 8 to 31 years with an average (SD) of 15.4 (6.2) years; the disease duration varied from 1 to 23 years with an average (SD) of 8.3 (5.8) years. Polyneuropathy index (PNI), expressed as a mean percentage of the normal for twelve indices over the four nerves obtained by motor conduction studies, was 93.9% on the average in patients with IDDM. The mean amplitude of CNAP obtained by intrafascicular microneurography was 417 microV. These results indicate that neuropathy in IDDM is milder than that in adult non-insulin dependent diabetes mellitus (NIDDM). The mean value of PNI decreased at a rate of 0.56% per year; the mean glycosylated hemoglobin (A1c) level was as high as 8.2 +/- 0.9%, findings consistent with those of the previous analysis of adult patients with NIDDM. The PNI value had a significant negative correlation with the duration of diabetes mellitus (p < 0.001) and with mean glycosylated hemoglobin (A1c) level (p < 0.01). CNAP amplitude had a tendency to correlate with duration of diabetes mellitus (p < 0.1). In patients with IDDM we can tell exactly when the disease occurred. Progression of neuropathy in juvenile IDDM was identical to that of adult NIDDM. Careful management of diabetes mellitus is of importance to prevent the progression of neuropathy. PMID- 9396029 TI - [Mitochondrial redox change in gerbil hippocampus before and after transient ischemia]. AB - To investigate the basis of neuronal vulnerability we studied mitochondrial redox changes in gerbil hippocampus before and after 5 minutes forebrain ischemia. The brain was frozen by in situ funnel freezing method, and grinned off coronally until exposure of hippocampus. Relative value of regional redox ratio (NAD+/NADH) was obtained from fluorescence signals of intrinsic fluorochromes, i.e., NADH (PN) and flavoproteins (Fp), using a high resolution fluorometer. We calculated a modified redox ratio MRR = FP/(Fp + PN). Each point is displayed in gray scales ranged 16 degrees corresponding to the MRR value of the point; black represents a low MRR value (reduced) and white represents a high value (oxidized). Pyramidal cell layers and the granule cell layers were seen as linear areas of high MRR. The stratum radiatum and stratum orience of the CA 1 subfield showed low MRR compared with other hippocampal regions. During ischemic period, MRR in all subfield of hippocampus had decreased but the decrease was more severe in CA 1 region than in another. Just after recirculation, MRR decreased transiently in dentate and CA 3 areas but was fully recovered in all hippocampal areas with the exception of CA 1 region, where the MRR decreased again 12 hours after recirculation. These results suggest that CA 1 area suffers more pronounced hyoxic condition (state V) than other less vulnerable regions during 5 minutes ischemia. The irreversible reduction of MRR in CA 1 area may result from continuing mitochondrial dysfunction, and this may cause lasting energy shortage in CA 1 neurons that eventually results in slowly progressive cell death. PMID- 9396030 TI - [Central nervous system involvement of leukemia and systemic lymphoma in children: CT and MR findings]. AB - The purpose of this paper is to retrospectively evaluate CT and MR findings of central nervous system (CNS) involvement of leukemia and systemic lymphoma in children. Over a 12-year period, sixty-five patients with leukemia and fifteen patients with systemic lymphoma underwent cerebral CT and/or MR imaging. Nine patients (11.3%) were diagnosed as CNS involvement of leukemia and lymphoma. The diagnostic criteria of CNS involvement were as follows; 1) Histological proof was confirmed by surgery, 2) Tumor cells were found in the cerebrospinal fluid examinations, 3) Increase in size of the lesion during observation without specific treatment, and 4) Response to the treatment for leukemia or lymphoma. All of nine patients fulfilled more than two criteria of 1)-4). The CT and MR abnormalities in these patients were correlated with the findings of histology, cerebrospinal fluid cytology, and/or treatment. The age of the patients ranged from 0 to 15 years old. They consisted of 6 boys and 3 girls. The CT examinations were performed before and after contrast administration. MR examinations were performed on a 1.5-T unit, and T1-weighted, T2-weighted, and proton density weighted images were obtained using spin-echo or fast spin-echo sequences. Tumor masses were present in seven with leukemia (acute lymphoblastic leukemia 4; acute myeloblastic leukemia 1; acute promyelocytic leukemia 1; acute monocytic leukemia 1), and in two with malignant lymphoma. On the CT scan, tumor masses were hyperdense with contrast enhancement. On the MR images, their signals were variable. In all of nine patients, tumor masses were contiguous with a meningeal surface. Postcontrast T1 weighted images were valuable in demonstrating meningeal infiltration. Tumoral hemorrhage was found in two patients. In a patient with tumor at the superior sagittal sinus, venous infarct was observed. CNS leukemic and lymphomatous masses are almost hyperdense on the CT and they are characteristically contiguous with a meningeal surface. MR imaging was valuable in demonstrating meningeal infiltration. Findings of CT and MR imaging, cerebrospinal fluid examinations, and response to the treatment are useful in the differentiation of CNS involvement of leukemia and lymphoma from other lesions such as infectious diseases and leukoencephalopathy. PMID- 9396031 TI - [Effect of arginine vasopressin (AVP) inhibitor in the inhibition of the peritumoral edema]. AB - We investigated the effect of RU599 Arginine Vasopressin (AVP) inhibitor on peritumoral edema of Wistar rats implanted with C6 glioma. C6 glioma was implanted into the right caudoputamen of 3 week-old rat's brain. Five weeks after the implantation, the changes of the brain water contents were measured by the dry-weight method. RU599 (1 mg/ml/kg body weight) was administered into Wistar rats via the tail vein every one hour (total 4 times), and the same dose of saline was used as control. Rats were sacrificed 1 hour after the last administration, and the brain of each rat was divided into 3 parts such as the anterior part without a tumor, the middle part with a tumor and the posterior part without a tumor. After that each part was separated into the right and left parts. The tumor was removed from the brain. The water contents of the brain were increased in the area around the tumor and the contralateral cerebral hemisphere excluding the anterior part. RU599 could reduce the increased water contents significantly in all areas except in the tumor itself and the area surrounding the tumor. Tumor-origin brain edema includes not only localized one but also diffuse one in the brain, and this study suggests RU599 has an effect of inhibiting the diffuse brain edema. PMID- 9396032 TI - [A Japanese family with Lesch-Nyhan syndrome resulting from a new point mutation in hypoxanthine-guanine phosphoribosyltransferase gene]. AB - Lesch-Nyhan syndrome is associated with complete deficiency of hypoxanthine guanine phosphoribosyltransferase (HPRT), characterized by hyperuricemia and severe neurological signs. The HPRT gene has been mapped to the q26 region on the long arm of the X-chromosome. We are taking care of a family of Lesch-Nyhan syndrome. A 14-year-old male was noted the growth disturbance at the age of 7 months and self-mutilation behavior characterized by compulsive biting of his lip and fingers at the age of 18 months. In 1987, at the age of 4, he was diagnosed as Lesch-Nyhan syndrome from neurologic signs and hyperuricemia (9.8 mg/dl). Neurological examination revealed mild mental and growth retardation, spasticity and hyperreflexia of lower extremities, choreoathetoid movements of extremities, and compulsive self-mutilation. The HPRT activity in erythrocytes of this patient was 0.02 nmol/min/mg hemoglobin (control value 1.76 +/- 0.06), and adenine phosphoribosyltransferase (APRT) activity was 1.08 nmol/min/mg hemoglobin (control value 0.43 +/- 0.06). Using polymerase chain reaction (PCR) method coupled with direct sequencing, we analyzed the nucleotide sequences of each exon from the genomic DNA as well as the entire HPRT coding region of the cDNA by RT PCR method. In the HPRT gene from the patient, a guanine to adenine substitution at base position 209 in exon 3 was identified, which resulted in a single amino acid substitution of glycine with glutamic acid at codon 70. The family studies indicated that his mother, sister and grandmother were heterozygotes. PCR restriction fragment length polymorphism (RFLP) utilizing Mnl I site which created by the mutation, was useful for detection of the mutant gene. We have identified a new missense mutation of the HPRT gene in a Japanese patient. This mutation was reported at the same codon as foreign mutants and mighty be indicative of a location of mutation activity in the HPRT gene. PMID- 9396033 TI - [Ibudilast prevents oligodendroglial excitotoxicity]. AB - Previously we have demonstrated that cells of oligodendroglial lineage express non-N-methyl-D-aspartate (NMDA) glutamate receptor (GluR) genes and are damaged by kainate induced Ca2+ influx via non NMDA GluR channels of the alpha-amino-3 hydroxy-5-methyl 4 isoxazole propionate (AMPA) type, representing oligodendroglial excitotoxicity. We here present the finding that ibudilast, which is used clinically for treat patients with asthma and cerebrovascular diseases, prevents oligodendroglia excitotoxicity. The oligodendrocyte like cells (OLC), differentiated from the CG-4 cell line established from rat oligodendrocyte-type 2 astrocyte (O-2A) progenitor cells, were exposed to 2 mM kainate for 24 h and cell death was evaluated by measuring activity of lactate dehydrogenase (LDH) released into the culture medium. Kainate induced cell death was prevented by 10 to 100 microM ibudilast, which increased intracellular cAMP. A 45Ca2+ influx study revealed that ibudilast attenuated kainate-induced Ca2+ influx. Inhibition of kainate-induced Ca2+ influx by ibudilast was decreased by H 89, a protein kinase A (PKA) inhibitor, but increased by okadaic acid, an inhibitor of phosphatase 1 and 2A. Therefore, we concluded that ibudilast prevented oligodendroglial excitotoxicity by a PKA-dependent phosphorylation process resulting in decreased kainate-induced Ca2+ influx. PMID- 9396034 TI - [A case of chronic encephalitis due to double infection with herpes simplex and measles viruses]. AB - In a healthy 49-year-old man, a decrease in job efficiency was noticed along with bizarre behavior. On admission, he was euphoric, childish, superficial and had increased libido. Neurological findings were normal. There were no abnormal findings on routine blood tests, hematochemistry or urine analysis. MRI showed no abnormal findings. However, single photon emission CT (SPECT) showed diffuse hypoaccummulation of tracer from the temporal to frontal regions. Lumbar puncture showed clear cerebrospinal fluid (CSF) with pleocytosis and an elevated protein level. Moreover, antibody IgG titers to herpes simplex virus (HSV) and measles virus were elevated, according to EIA [serum HSV -1,202.2x, measles virus 47.1x: CSF HSV-116.1x, measles virus 9.9x]. The ratio of serum to CSF antibody titers of HSV and measles virus were 12.5 and 4.75, respectively. The antibody index values of HSV and measles virus IgG titers were 8.42 and 22.22. The ratio of albumin was 105.7. Chronic, progressive HSV encephalitis is rare, and there have been very few reports of encephalitis due to double infection by HSV and another virus. Our patient was diagnosed as having encephalitis due to double infection with HSV and measles virus, because the ratio of serum to CSF antibody titers was less than 20 and the antibody index values were over 1.91. Moreover, since the IgG index was elevated and the ratio of albumin was not low, it was suggested that the blood brain-barrier had not been disrupted, and antibodies were being produced chronically in the medullary cavity. Hyperaccummulation of tracer on SPECT studies has been reported in the early stages of HSV encephalitis. In our case, while CT and MRI showed no abnormal findings, SPECT showed diffuse hypoaccummulation. SPECT appears to be a useful tool in the diagnosis of this disorder. In case of chronic, progressive personality change in middle-aged adults, we must be aware of double virus infection of the brain as a possible causal factor. PMID- 9396035 TI - [Myotonic dystrophy with marked megacolon: report of a case]. AB - A 45-year-old woman was incidentally suspected to have megacolon. Chest X-rays showed elevated left diaphragm due to colonic gas, and the heart was deviated to the midline. Barium enema revealed marked dilation of the sigmoid colon, confirming the diagnosis of megacolon. Maximal diameter of the sigmoid colon was 23 cm, but she had no gastrointestinal symptoms. During the work up for megacolon, the presence of myotonic dystrophy was suspected. She had hatchet face, but was not bald. Muscles of the neck and extremities were slightly atrophic. There was percussion myotonia of the tongue and both hands, and grip myotonia of the hands. Laboratory examinations showed impaired glucose tolerance and low level of serum IgG. EMG showed myotonic discharges and myopathic units in the limbs. Brain CT imaging revealed a thick skull. Cases of myotonic dystrophy associated with marked megacolon are rare in Japan. Megacolon presents a high risk for ileus, volvulus, and rupture, and myotonic dystrophy is associated with a high operative and anesthesic risk. Megacolon, therefore, is an important complication to look for in the management of myotonic dystrophy. PMID- 9396036 TI - [A case of midbrain infarction with ipsilateral hand tremor]. PMID- 9396037 TI - [Venous angioma]. PMID- 9396038 TI - [(Neurological CPC-59). A 65-year-old man with a history of gastric cancer who presented progressive loss of vision, memory loss and consciousness disturbance]. AB - We report a 65-year-old man with progressive loss of vision and consciousness disturbance. The patient was well until his age of 63 when he was found to have a gastric cancer. He was treated by the tumor resection and chemotherapy; he was apparently well, but hepatic metastases were found in the next year (1996). In June, 1996, he noted an onset of blurred vision more on the left. He was admitted to the ophthalmology service of our hospital on July 14, 1996. His vision was 0.8 on the right and 0.15 on the left. He was treated with oral prednisolone with slight improvement. He was also found to have IgM kappa-type monoclonal gammopathy; Bence-Jones protein was positive and a bone marrow aspiration revealed that approximately 10% of bone marrow cells were atypical plasma cells. His vision had progressively got worse and he became blind by the end of October 1996. A chest X-ray and cranial CT scan revealed multiple abnormal nodular densities. In the middle of November 1996, he became confused, disoriented and agitated. His mental symptoms had progressively became worse, and a neurologic consultation was asked on December 10, 1996. Neurologic examination revealed that he was somnolent with decreased attention to his surroundings. He showed marked disorientation and memory loss. Higher cerebral functions appeared intact. He was able to recognize only light and dark. Pupils were moderately dilated with very sluggish light reflex remained. Vertical gaze was moderately restricted and horizontal nystagmus was noted upon left and right lateral gaze. The remaining of the neurologic examination were unremarkable. General physical examination revealed hepatosplenomegaly; the liver was palpable by 3 cm below the right costal margin. Laboratory examination revealed anemia (Hb10.1 g/dl) and thrombocytopenia (43,000/microliter). A cranial CT scan and MRI revealed a mass lesion in involving the chiasmatic and bilateral hypothalamic areas. The tumor showed intense homogeneous enhancement after Gd-DTPA infusion. The patient was treated with dexamethasone and radiation. After 9 Gray radiation, he showed deterioration in the sensorium; a cranial CT scan revealed a hydrocephalus of the right ventricle with the midline shift towards left. The radiation was discontinued. The subsequent clinical course was complicated by aspiration pneumonia and thrombocytopenia. He expired on January 4, 1997. The patient was discussed in a neurological CPC and the chief discussant arrived at the conclusion that the patient had systemic malignant lymphoma with metastasis to the brain judging from the characteristics of MRI and CT findings. Opinions were divided between malignant lymphoma and metastatic brain tumor. Post-mortem examination revealed plasmacytoid lymphocytic infiltration in the bone marrow. Immunologically, these cells were positive for IgM and kappa-type light chain. These plasmacytic infiltrations were seen in the lungs and lymph nodes. These findings were consistent with the diagnosis of Waldenstrom's macroglobulinemia. In the liver metastatic cancer tissues were seen; microscopic pictures were essentially similar to those of resected gastric cancer. No local recidive was noted in the stomach. In the central nervous system, a necrotic tissue was involving the hypothalamic area bilaterally; no clear neoplastic cells were found, however, lymphocytic and plasmacytic infiltrations were seen in the perivascular space. In the optic nerves, loss of myelin and axons were seen. These findings most likely mass formation from macroglobulinemia which underwent necrotic change after radiation. Mass formation in the brain is rare for Waldenstrom's macroglobulinemia, although it has been reported. The relation between gastric cancer and macroglobulinemia in this patient is unclear. PMID- 9396040 TI - Tamoxifen alters the localization of F-actin and alpha 5/beta 1-integrin fibronectin receptors in human endometrial stromal cells and carcinoma cells. AB - We have investigated F-actin and the integrin fibronectin receptor as possible targets of tamoxifen (TAM) signaling in a cell-based model of the endometrium. Normal human endometrial stromal cells and RL95-2 human endometrial adenocarcinoma cells were treated for 1 h with TAM, a known antagonist of protein kinase C (PKC), or with staurosporine or HA1004, two broad-spectrum protein kinase antagonists capable of inhibiting PKC and PKA, respectively. We utilized fluorescein-phalloidin and confocal microscopy to visualize the cellular distribution of F-actin. Normal stromal cells and RL95-2 cells differed in the arrangement of F-actin in control cells and in their response to TAM. In control stromal cells, actin stress fibers were well organized throughout the cell, but in RL95-2 cells, they were disorganized and present mainly at the cell periphery. F-actin in RL95-2 cells treated with TAM (0.1 and 1.0 microM) or with staurosporine (0.7 and 7.0 nM) exhibited a reorganization into stress fibers consistent with a more stationary phenotype. In contrast, TAM- or staurosporine treated normal stromal cells exhibited an increase in the amount of organized F actin. Interestingly, in normal stromal cells treated with staurosporine but not TAM or HA1004, these F-actin fibers appeared to terminate in dense plaques proximal to the plasma membrane. The alpha 5/beta 1 integrin fibronectin receptor mediates between the extracellular matrix and the actin cytoskeleton. TAM induced clustering of the fibronectin receptor at the plasma membrane in normal stromal cells, but not in carcinoma cells. This study supports the importance of plasma membrane-cytoskeletal protein interactions in the response of normal and carcinoma cells to TAM. PMID- 9396039 TI - Long-term resistance to HIV infection in vertical HIV infection: cytokine production, HIV isolation, and HIV phenotype define long-term resistant hosts. AB - We analyzed immunologic (CD4 and CD8 slopes; interferon-gamma, interleukin-2, interleukin-10, and chemokines production; concentration of IgE; beta 2 microglobulin) and virologic (p24; HIV isolability and phenotype; plasma viremia) parameters in HIV vertically infected children > or = 8 years of age without disease progression or mild symptoms and an absolute CD4+ count > or = 500/microliter with CD4+ percentage > or = 25%. The results were compared to those of two control groups: (1) slow progressors, children > or = 8 years of age with moderate symptomatology and/or moderate CD4 depletion, and (2) progressors, children > or = 8 years of age with severe clinical disease and/or severe CD4 depletion. Pediatric long-term resistant hosts were characterized by higher production of interleukin-2 and interferon-gamma and lower production of interleukin-10, normal concentration of IgE, HIV isolates with a non-syncytium inducing phenotype, and lower plasma viremia. This condition was not associated with the concentration of beta 2-microglobulin, p24, and chemokines, or with HIV isolability. The IL-10/IL-2 ratio best correlated with both CD4 counts and disease progression. Thus, vertically infected children showing resistance to disease progression are immunologically and virologically distinct from those in whom progressive HIV infection is observed. PMID- 9396041 TI - Stage I and stage II infiltrating ductal carcinoma of the breast analyzed for chromosome 8 copy number using fluorescent in situ hybridization. AB - We previously reported the results of 30 informative samples (from a total of 34 specimens gathered) of archival breast cancer tissue, including infiltrating ductal carcinoma (NOS), ductal carcinoma in situ, lobular carcinoma, papillary carcinoma and benign lesions of the breast. The study was conducted using fluorescent in situ hybridization (FISH) and a chromosome 8 alpha-satellite probe. Subsequently, a total of 34 cases of infiltrating ductal carcinoma of the breast (NOS, 17 cases stage I and 17 cases stage II) were studied, again using interphase cytogenetics. The aim of the present study is to confirm and extend the results of our initial study of stage I and stage II disease. Towards this end, 36 additional specimens of formalin-fixed paraffin-embedded breast cancer tissue have been analyzed cytogenetically under blinded conditions for the frequency of abnormal chromosome 8 copy numbers using FISH and the previously described protocol optimized for our laboratory. Of these, 18 were stage I and 18 were stage II. The frequency of trisomy 8 among stage I tumors was found to be 28% (5 out of 18). The frequency of trisomy 8 among stage II tumors was found to be 61% (11 out of 18). These results, while less striking, are consistent with those reported in our initial study of stage I and stage II disease, where the frequencies of trisomy 8 among stage I and stage II tumors were 24% (4 out of 17) and 82% (14 out of 17). These results not only establish that chromosome 8 trisomy is a recurrent finding in breast cancer, but also confirm that a higher frequency of trisomy 8 was observed with a higher clinical stage (stage II) than with a lower stage (stage I). It will be of interest to extend the findings in stage I and stage II breast cancer to other stages as well. PMID- 9396042 TI - Luzindole, a melatonin receptor antagonist, suppresses experimental autoimmune encephalomyelitis. AB - Melatonin has immune-enhancing effects and can exacerbate autoimmunity. Pinealectomy or light exposure, which suppress melatonin, inhibit T cell autoimmunity. To investigate the involvement of melatonin in experimental autoimmune encephalomyelitis (EAE), a T-cell-mediated autoimmune demyelinating disease, we tested the effect of luzindole, a melatonin receptor antagonist, on EAE. Luzindole-treated mice did not develop EAE after immunization with spinal cord homogenate, whereas control mice developed EAE. This study suggests that pharmacological inhibition of the immunoenhancing effects of melatonin may prevent autoimmune demyelination. PMID- 9396044 TI - 'Autoantibody dominance' pattern following idiotypic manipulation of naive mice by immunization with anti-U1RNP antibodies. AB - OBJECTIVE: To study the immune response and clinical findings in mice immunized with different epitope-specific anti-U1RNP antibodies purified from the sera of mixed connective tissue disease (MCTD) patients with various clinical manifestations. METHODS: BALB/c mice were immunized with anti-U1RNP-IgG preparations from 3 patients with MCTD. Group 1 was immunized with U1 70 kD A positive IgG, group 2 with U1 70 kD-negative, U1A, U1C, B-B'-positive IgG and group 3 with U1 70 kD, U1A, U1C-positive IgG. The induced autoantibody response in the mice was studied by ELISA and immunoblots and the clinical findings of MCTD in humans were assessed. RESULTS: Immunoblot assays showed that mice immunized with different human anti-U1RNP antibodies developed predominantly autoantibodies directed against U1 68-70 kD epitope. This 'autoantibody dominance' pattern was not associated with clinical findings. CONCLUSIONS: The restricted murine autoimmune response may provide clues to the diversified autoantibody production in autoimmune diseases and explain in part the changing patterns of clinical findings in individuals with MCTD. PMID- 9396043 TI - Overexpression of protein tyrosine kinases in human esophageal cancer. AB - Using a PCR-based cloning technique, we isolated a series of protein tyrosine kinases (PTKs) expressed in a cell line of esophageal squamous cell carcinoma. Sequence analysis revealed 10 different kinds of PTKs of the receptor type [epidermal cell growth factor receptor, insulin-like growth factor I receptor, fibroblast growth factor receptor 4, eck, erk, discoidin domain receptor (DDR)/trkE/cell adhesion kinase (Cak), HEK2, HEK8, axl and sky] and one PTK of the nonreceptor type (tyk2). Subsequently, we examined the expression of the transcripts of these 11 genes in paired samples of normal and carcinomatous esophageal tissues obtained from 12 cases of esophageal cancer. We found that all 11 gene transcripts were expressed in both carcinomatous and normal tissues, and 6 of them were significantly overexpressed in carcinomatous tissues relative to adjacent normal tissues. Among these, the magnitude of mRNA expression of DDR/trkE/Cak PTK was positively correlated with the proliferative activity of carcinoma cells, but not with their degree of differentiation. Immunohistochemically, DDR was expressed in both normal and cancerous esophageal cells. The intensity of the expression was higher in cancer than normal tissue. In addition, we confirmed the expression of two isoforms of DDR/trkE/Cak in normal and cancerous esophagus. Our study suggests that DDR/trkE/Cak plays an important role in the regulation of proliferation of esophageal cancer. PMID- 9396045 TI - HLA-DQB1 markers associated with human immunodeficiency virus type I disease progression. AB - In a previous investigation, we demonstrated that certain human leukocyte antigens (HLA) may be associated with human immunodeficiency virus type I (HIV-1) infection or protection from infection among regional African Americans and Caucasians. We demonstrated that HLA-DQB1*0605 was associated with a possible increased risk of susceptibility to infection in African Americans and that DQB1*0602 was associated with a possible increased risk of infection in Caucasians. The present study was designed to demonstrate possible HLA associations with HIV-1 disease progression and AIDS in regional African American and Caucasian populations. To differentiate rapid from slow progressors, immune parameters of the HIV-1-positive patient population were monitored over a mean follow-up period of 23 +/- 2 months for African Americans (n = 30) and 25 +/- 5 months for Caucasians (n = 22). To determine significance, HLA allele frequencies among rapid progressors were compared to those of slow progressors, separated by race. Results were analyzed by chi 2 analysis, with Fisher's exact test where applicable, linear logistic regression and Kaplan-Meier survival analysis. In the HIV-1-positive African American group, a better prognosis was associated with HLA DQB1*0602. In the HIV-1-positive Caucasian group, HLA-DQB1*0302 was associated with rapid HIV disease progression, but no marker was associated with a more favorable prognosis. PMID- 9396047 TI - Translocation 2;19 in a patient with probable relapsed acute myeloid leukemia. AB - We report the cytogenetic and hematopathologic results from a patient diagnosed with acute myeloid leukemia. Although the initial specimen revealed an apparently normal male karyotype, a translocation, t(2;19)(q21;p13), was detected in the second specimen. It is not clear whether this was a primary or secondary and possibly chemotherapy-induced abnormality. In an extensive search of the recent medical literature database (Medline, 1966 to the present; CancerLit, 1983 to the present, MDX Health Digest, 1988 to the present; HealthSTAR, 1975 to the present, and CINAHL, 1982 to the present), we found no previous report of this specific translocation. This case is of interest not only because of its cytogenetic rarity and its unique clinical features, but also because of the fact that this patient worked in construction management, performing offshore drilling in oil fields for several years, and also worked with plastics and polymer film for about 4 years, although this past history of possible genotoxic exposure may or may not be of relevance. In addition, it is also of interest that one of the translocation breakpoints, 19p13, is apparently identical to that found in the 1;19 translocation associated with pre-B cell acute lymphocytic leukemia. PMID- 9396046 TI - Mature CD4 single positive thymocytes in human Thymoma: T cells may differentiate in the thymic epithelial cell tumor. AB - Human thymoma, which is occasionally associated with autoimmune disease, is a thymic epithelial cell tumor and often contains a large number of lymphocytes. In a previous study, we have shown that a proportion of CD4 single positive T cells in human thymomas lack CD3, suggesting immaturity. In this study, we focused on the rest of the CD4 single positive T cells in thymomas that expressed CD3/TcR alpha beta and investigated the maturity of single positive T cells by analyzing lymphocyte surface antigens and the cells' proliferative response to a mitogen. CD4 single positive cells that expressed CD3 or TcR alpha beta also expressed CD69 and had probably undergone positive selection in the tumor. Further, isolated CD4 or CD8 single positive cells from the thymomas responsed to a mitogen although at lower levels than the corresponding single positive cells in the peripheral blood. These results indicate that thymomas contain single positive T cells which have mature phenotype and proliferative ability, and suggest that T cells may differentiate in thymoma. PMID- 9396048 TI - Characterization of myomodulin-related peptides from the pulmonate snail Helix aspersa. AB - Three myomodulin-related peptides--pQLSMLRLamide, PMSMLRLamide, and SLGMLRLamide- have been purified and sequenced from extracts of whole snails. The level of immunoreactive myomodulin was shown by HPLC and RIA to be widely distributed among 26 different snail tissues, with the highest levels (higher even than those in the central ganglia) occurring in certain male reproductive organs. Synthetic pQLSMLRLamide modified either the spontaneous rhythmic activity or the resting tone of several isolated muscular organs: the aorta, ventricle, upper gut, epiphallus, flagellum, and spermatheca; but the retractor muscles of the pharynx, penis, and tentacle were unaffected. PMID- 9396049 TI - Neutral endopeptidase-24.11 (NEP)-like activity in molluscan hemocytes. AB - Using a spectrofluorimetric procedure, we found that the plasma membrane from hemocytes of two freshwater snails, Planorbarius corneus and Viviparus ater, shows neutral endopeptidase-24.11 (NEP)-like activity. Moreover, the addition of ACTH(1-24) to the hemolymph provokes an increase in NEP-like activity. This increased NEP-like activity is blocked by phosphoramidon, a potent inhibitor of mammalian NEP. These findings suggest that this peptidase has been well conserved in the course of evolution and plays an important role in immune-neuroendocrine mechanisms. PMID- 9396050 TI - Adrenomedullin binds with high affinity, elevates cyclic AMP, and stimulates c fos mRNA in C6 glioma cells. AB - The effects of adrenomedullin (ADM) on C6 glioma cells were investigated. [125I]ADM bound with high affinity (Kd = 24 nM) to a single class of sites (Bmax = 36,000/cell) in C6 cells. Specific [125I]ADM binding was inhibited with high affinity by ADM (IC50 value of 10 nM) but not ADM(22-52) or pro-adrenomedullin N terminal 20 peptide (PAMP). By RT-PCR, ADM receptors were detected in C6 cells. ADM elevated cAMP (ED50 value of 10 nM) whereas PAMP and ADM(22-52) did not. ADM stimulated transiently c-fos mRNA in a concentration-dependent manner. Monoclonal antibody G6, which neutralizes ADM, significantly inhibited C6 proliferation and decreased the ability of ADM to elevate c-fos mRNA. These data suggest that ADM is a regulatory peptide of C6 cells. PMID- 9396051 TI - Production and secretion of adrenomedullin from glial cell tumors and its effects on cAMP production. AB - The expression of adrenomedullin (ADM) and its mRNA was studied in human glial cell tumors and cultured glioblastoma cell lines, T98G and A172. Northern blot analysis showed that ADM mRNA was expressed in all brain tumors examined (three anaplastic astrocytomas and two glioblastomas multiforme) and in the glioblastoma cell lines. Immunoreactive (IR-) ADM was detectable in these brain tumors by radioimmunoassay (0.31-2.0 pmol/g wet weight), except for one anaplastic astrocytoma. Reverse phase high performance liquid chromatography of the tumor extracts showed a single peak eluting in the position of ADM-(1-52). IR-ADM concentrations in the cultured media of T98G cells were 205.5 +/- 8.4 fmol/10(5) cells/24 h (mean +/- SEM, n = 5). Treatment of T98G cells with interferon gamma or interleukin 1 beta increased the expression levels of ADM mRNA and the IR-ADM concentrations in the cultured media, whereas tumor necrosis factor alpha decreased them in a dose-dependent manner. Treatment with synthetic ADM-(1-52) (10(-8) or 10(-7) mol/l) increased the cAMP concentrations in the cultured media of T98G cells. These findings suggest that ADM is secreted from glial cell tumors and is related to the pathophysiology of these tumors. PMID- 9396052 TI - Specific adrenomedullin binding sites in the human brain. AB - Binding sites for adrenomedullin in human brain were investigated and characterized by radioligand binding. Specific binding sites for adrenomedullin were present in every region of human brain (cerebral cortex, cerebellum, thalamus, hypothalamus, pons and medulla oblongata) obtained at autopsy. Despite the homology with calcitonin gene-related peptide (CGRP), CGRP was a poor inhibitor of [125I]adrenomedullin binding (IC50 > 1 microM) compared with adrenomedullin(1-52) (IC50 = 1.2 +/- 0.5 nM, mean +/- SEM, n = 3). Three adrenomedullin fragments, adrenomedullin(1-12), adrenomedullin(22-52), and adrenomedullin(13-52), were also poor inhibitors of the binding (IC50 = 0.3 microM), suggesting that the whole molecule of adrenomedullin(1-52) is required for binding to the receptor. Scatchard plots of [125I]adrenomedullin binding in human brain (cerebral cortex) gave a dissociation constant of 0.17 +/- 0.03 nM and maximal binding of 99.3 +/- 1.9 fmol/mg protein (n = 5). These findings suggest that specific adrenomedullin binding sites that differ from the CGRP receptors exist in human brain. This indicates a possible novel neurotransmitter/neuromodulator role for adrenomedullin in human brain. PMID- 9396053 TI - Neurobehavioral development of neonatal mice following blockade of VIP during the early embryonic period. AB - Previous work has shown that blockade of VIP function in the early postimplantation embryo results in growth retardation and microcephaly. In the present work, the neurobehavioral development of neonatal mice was examined following treatment of dams with a VIP antagonist during this period. Inhibition of VIP functions during early embryogenesis impaired the performance of 5 of 10 developmental behaviors. These behaviors included developmental milestones (first appearance of ear twitch and eye opening) and complex motor behaviors (negative geotaxis, surface righting, and air righting). The retardation of neurobehavioral development produced by inhibition of VIP action indicates that this peptide is important to the progression of embryonic development. PMID- 9396054 TI - Vasoactive intestinal polypeptide (VIP) innervation of rat spleen, thymus, and lymph nodes. AB - In the thymus, VIP-positive (+) fibers were found in the capsular/septal system, cortex, and medulla. In the spleen, VIP+ nerves coursed along large arteries and central arterioles, and in the white pulp, venous/trabecular system, and red pulp. Splenic VIP innervation was more robust in Long-Evans hooded rats than in Fischer 344 rats. VIP+ nerves in mesenteric lymph nodes were found in the cortex, and along the cortical vasculature and medullary cords. No VIP innervation was observed in popliteal lymph nodes. Immunocytes also were VIP+, suggesting that both neural and cellular synthesis of VIP contributes to VIP concentration in lymphoid organs. Surgical sympathectomy did not alter splenic or thymic VIP content, respectively, and VIP innervation of these organs was not altered, suggesting an origin for VIP+ nerves other than the sympathetic nervous system. PMID- 9396055 TI - Vasoactive intestinal peptide stimulates p59fyn kinase activity in murine thymocytes. AB - The neuropeptide VIP has immunomodulatory properties, including the inhibition of cytokine production (IL-2, IL-4, and IL-10) in T lymphocytes stimulated through their TCR. The transduction pathways involved in the inhibitory effect of VIP on IL-2 expression are not known. Here we investigate the effect of VIP on the T cell-specific protein tyrosine kinases p56lck and p59fyn in resting and stimulated thymocytes. VIP does not affect lck or fyn activity in stimulated thymocytes and does not alter the general pattern of cellular tyrosine phosphorylation. However, VIP stimulates p59fyn, but not p56lck, kinase activity in resting thymocytes. The effect is dose dependent, exhibits a specific time course, and is reproduced by other cAMP-inducing agents such as forskolin, prostaglandin E2, and 8-bromo-cAMP, suggesting that cAMP may function as the intracellular mediator. PMID- 9396056 TI - Pretreatment with neurokinin substance P but not with cholecystokinin-8S can alleviate functional deficits of partial nigrostriatal 6-hydroxydopamine lesion. AB - The neuropeptide substance P (SP) has been implicated in the control of various neuro-behavioral functions including reinforcement and learning processes. It also exerts neurotrophic and regenerating effects in vitro and in vivo. A previous study indicated a potential therapeutic effect of SP in rats with partial 6-hydroxydopamine lesions of the nigrostriatal dopamine system when SP was administered after the lesion. The purpose of the present study was to determine whether prelesion treatment with SP would also interact with the effects of unilateral 6-hydroxydopamine lesion of the substantia nigra. Thus, SP (50 micrograms/kg) was administered i.p. on 8 consecutive days prior to unilateral lesion of the substantia nigra. Furthermore, we investigated the effects of prelesion treatment with cholecystokinin-8S (CCK; 1 microgram/kg), another neuropeptide, which is closely related to dopaminergic neurons, and which also can have neurotrophic and neuroprotective functions. Our results show that animals with partial neostriatal dopamine depletions (residual dopamine levels of more than 10%) did not show turning asymmetries when pretreated with SP, whereas animals pretreated with vehicle exhibited an initial ipsiversive asymmetry from which they recovered. In contrast, behavioral asymmetries were most pronounced in animals which had been pretreated with CCK. These peptide treatments did not affect the degree of neostriatal dopamine depletion; however, dihydroxyphenylacetic acid/dopamine ratios were enhanced in the neurostriatum of animals with partial dopamine damage after SP- and CCK-pretreatment, and in the ventral striatum of SP-pretreated animals. These data provide evidence that prelesion treatment with SP, but not with CCK, can alleviate functional deficits induced by a partial nigro-striatal dopamine lesion. This effect may be related to enhanced ventral striatal dopamine activity and/or to the peptide's known effects on learning, motivation, and emotion. PMID- 9396057 TI - Gastric and brain stem, effects of substance P on nucleus tractus solitarius unitary responses. AB - Subdiaphragmatic vagally evoked unitary responses were recorded in the medial subnucleus of the nucleus tractus solitarius in an in vitro neonatal rat brain stem-gastric preparation. Substance P was applied to the gastric compartment and/or the brain stem compartment of the bath chamber to evaluate the peripheral gastric and central brain stem effects of the peptide on the nucleus tractus solitarius unitary activity. After substance P application to the gastric or brain stem compartment, a concentration-related change in the nucleus tractus solitarius unitary activity was observed. The gastric effects of substance P, at a concentration of 30 nM (minimum concentration for maximum effect), produced a 31 +/- 6.5% (mean +/- SD) increase in neuronal discharge frequency compared to the control recording. Both the proximal and the distal stomach were important in the gastric effect of the peptide on subdiaphragmatic vagally evoked nucleus tractus solitarius unitary responses. The brain stem effects of substance P. at a concentration of 10 nM (minimum concentration for maximum effect), induced a 52 +/- 11.3% increase in discharge frequency. Application of the peptide into both the gastric and brain stem compartments resulted in a subadditive response of 73 +/- 5.9%. This suggest that, peripherally and centrally, there are two different mechanisms of substance P activation. Our results suggest that substance P may play a role in regulating the ingestive process in neonates. PMID- 9396058 TI - Orthodromic activation of peptidergic cells in the medial amygdala. AB - Electrical recordings from vasopressin-containing cells in the medial amygdala were obtained. Electrical stimulation of one major afferent structure, the accessory olfactory bulb, invariably elicited single unit discharge in the peptidergic cells and set up a field potential indicating widespread excitation in the structure. Pheromonal stimuli, normally borne into the brain by the accessory olfactory bulb, were ineffective in activating the medial amygdala. These results in combination with preexisting research suggest that the accessory olfactory bulb is an important influence, but not the only influence, on the activity of the peptidergic cells. PMID- 9396059 TI - Facilitation of AVP(4-8) on gene expression of BDNF and NGF in rat brain. AB - In situ hybridization and Northern blot assay were used to evaluate the effects of exogenous AVP(4-8) on the transcription of mRNAs for nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and neurotrophin 3 (NT-3) in the adult rat brain. NGF and BDNF expression was found to be significantly enhanced by AVP(4-8) administration in the cerebral cortex and hippocampus, but NT-3 expression was not changed. In the same conditions, behavior-active arginine vasopressin (AVP) showed a small effect and its behavior-inactive homologue, oxytocin did not. Our results suggest that selective regulation of neurotrophin gene expression by the peptides may be responsible for its memory-enhancing function. PMID- 9396060 TI - Naltrexone, naltrindole, and CTOP block cocaine-induced sensitization to seizures and death. AB - I.c.v. injection for 9 days of either naltexone (NTX) (5, 10, 20, 40 micrograms/rat) or a selective mu peptide (CTOP) (0.125, 0.25, 0.5, 1, 3, 6 micrograms/rat) or delta (naltrindole) (NLT) (5, 10, 20 micrograms/rat) subtype opioid receptor antagonist affected sensitization to cocaine (COC) (50 mg/kg, i.p.) administered 10 min after. NTX (5 and 40 micrograms/rat), NLT (10 and 20 micrograms/rat), and the peptide CTOP (0.25-0.5 microgram/rat) attenuated seizure parameters (percent of animals showing seizures, mean score and latency) in a day related manner. The DD50 (days to reach 50% of death) value for COC was 2.69, whereas it was 9.67 and 7.27 for NTX 5 and 40 micrograms/rat, 8.59 for NLT (10 micrograms/rat), and 6.11, 5.95, and 4.30 for CTOP (0.25, 0.5, and 1 microgram/rat respectively). These findings suggest a concurrent involvement of mu- and delta-opioid receptor subtype in COC-induced sensitization to toxic effects. PMID- 9396061 TI - Decreases in systemic arterial and hindquarters perfusion pressure in response to nociceptin are not inhibited by naloxone in the rat. AB - Nociceptin, the endogenous ligand for the ORL1 receptor, has been shown to decrease systemic arterial and hindquarters perfusion pressures in the rat. The present study was undertaken to determine if decreases in systemic arterial and hindquarters perfusion pressures, in response to nociceptin, are mediated by a naloxone-sensitive mechanism. Injections of nociceptin decreased systemic arterial and hindquarters perfusion pressures in a dose-related manner. The decreases in systemic arterial and hindquarters perfusion pressure in response to nociceptin were not altered by the administration of naloxone in a dose of 2 mg/kg i.v. Met-enkephalin decreased systemic arterial and hindquarters perfusion pressures and responses to the opioid receptor agonist were significantly reduced by naloxone, whereas decreases in systemic arterial pressure in response to the nitric oxide donor, DEA/NO, were not altered. The results of the present study show that decreases in systemic arterial and hindquarters perfusion pressure in response to nociceptin are not mediated by a naloxone-sensitive mechanism in the rat. PMID- 9396062 TI - Evaluation of chronic opioid receptor antagonist effects upon weight and intake measures in lean and obese Zucker rats. AB - Body weight and food intake are significantly reduced in rats during development of dietary obesity following chronic central administration of mu (beta funaltrexamine, BFNA), mu1 (naloxonazine), kappa1 (nor-binaltorphamine, NBNI), delta1 ([D-Ala2,Leu5,Cys6]-enkephalin, DALCE) and delta2 (naltrindole isothiocyanate, NTII) opioid receptor subtype antagonists. In contrast, rats made obese by maintainance on a 'cafeteria' diet failed to display weight loss following chronic mu1 receptor antagonism. To test the hypothesis that chronic administration of opioid antagonists are less effective in controlling intake and weight in obese animals, the present study assessed whether chronic, central administration of either BFNA (20 micrograms), naloxonazine (50 micrograms), NBNI (20 micrograms), DALCE (40 micrograms) or NTII (20 micrograms) altered weight and intake in lean and obese Zucker rats over seven days. Body weight was reduced following chronic mu (lean: 42 g; obese: 49 g), mu1 (lean: 71 g; obese: 38 g), kappa1 (lean: 30 g; obese 14 g), delta1 (lean: 43 g; obese: 22 g) or delta2 (lean: 37.5 g; obese: 36 g) antagonism. Overall food intake was reduced following chronic mu (lean: 8.8 g; obese: 16.1 g), mu1 (lean: 12.6 g; obese: 17.0 g), kappa1 (lean: 6.5 g; obese 7.0 g), delta1 (lean: 9.7 g; obese: 11.1 g) or delta2 (lean: 9.4 g; obese: 14.3 g) antagonism. Therefore, both lean and obese Zucker rats display weight loss and reduced intake following chronic central administration of opioid receptor subtype antagonists. PMID- 9396063 TI - Modulation of dopamine receptor agonist-induced rotational behavior in 6-OHDA lesioned rats by a peptidomimetic analogue of Pro-Leu-Gly-NH2 (PLG). AB - The present study was undertaken to determine if the previously reported in vitro interactions of the Pro-Leu-Gly-NH2 (PLG) peptidomimetic analogue 3(R)-[(2(S) pyrrolidinylcarbonyl)amino]-2-oxo-1-pyrrolidineacet amide (PAOPA) with the dopaminergic system could be exhibited in an in vivo animal model using 6 hydroxydopamine (6-OHDA)-lesioned rats. In this model, PAOPA was found to potentiate the contralateral rotational behavior induced by either apomorphine or L-DOPA. PAOPA was 100-fold more potent than PLG, and produced a fourfold greater response than PLG when administered i.p. PAOPA also potentiated contralateral rotations induced by SKF-38393 and quinpirole. In summary, the results of this study indicate that PAOPA, a conformationally constrained peptidomimetic analogue of PLG, can modulate dopaminergic activity in vivo with higher potency and efficacy than PLG. PMID- 9396064 TI - The detection of thyrotropin-releasing hormone (TRH) and TRH receptor gene expression in Siberian hamster testes. AB - Thyrotropin-releasing hormone (TRH) from the hypothalamus is the major regulator of TSH synthesis and secretion. Most recently, TRH and TRH receptors (TRH-R), as well as their mRNAs, have been identified in rat testis. To expand our knowledge on the testicular TRH and TRH receptor gene expression in different species, in the present study the mRNA levels of testicular TRH and TRH-R were investigated in Siberian hamsters. To further localize the cellular sites of the gene expression, the animal model was treated with a single injection of ethylene dimethane sulfonate (EDS) (i.p., 80 mg/kg body weight), a compound known as to specifically eliminate testicular Leydig cells. The elimination of Leydig cells induced by EDS treatment was confirmed by histological studies of the testis sections and by serum hormonal analyses, which showed a dramatic reduction of serum testosterone (T) levels and significantly elevated serum LH concentrations. Messenger RNA levels of TRH and TRH-R in the testes were determined by Northern blot analyses quantitated with densitometry scanning. The results showed that specific TRH-R mRNA, 3.8 kb in size, was identified in Siberian hamster testes and the mRNA levels were significantly elevated in the EDS-treated testes compared to the controls (p < 0.01). Testicular TRH mRNA was also detected; however, no significant differences in TRH mRNA levels were found between EDS treated and control groups. The size of TRH mRNA was characterized as about 1.2 kb in hamster testes, which was smaller than that observed in the rat hypothalamus (1.6 kb) and in the rat testis (2.0 kb). Further studies by RNase H digestion revealed the presence of smaller TRH transcripts in the hamster testes than those in the rat testis. No hybridization signal for TRH mRNA was detected by RNase protection assay, when a rat TRH riboprobe was applied to hamster testis RNA, suggesting the limited homology of TRH gene sequences between these two species. Our results demonstrate that both TRH and TRH-R genes are expressed in Siberian hamster testes, and a significant increase of TRH-R mRNA levels occurs in the Leydig cell eliminated hamster testes. Unlike the rat testicular TRH mRNA mainly detected in Leydig cells, in hamster TRH mRNA could also be detected in other testicular compartment. PMID- 9396065 TI - Comparative antipsychotic profiles of neurotensin and a related systemically active peptide agonist. AB - Several lines of evidence have shown that neurotensin can modulate dopamine neurotransmission. It has been suggested that neurotensin has potential antipsychotic activity because it reduces dopaminergic activity preferentially in the nucleus accumbens. In the present study, the effects of neurotensin and NT1 (N alpha Me-Arg-Lys-Pro-Trp-TIe-Leu or Eisai hexapeptide), a metabolically stable and systemically active neurotensin agonist, were examined in several models of antipsychotic activity and side effect liability in mice; analgesic and hypothermic effects of both compounds also were determined. Up to high doses, neurotensin (5.0 and 10.0 micrograms, i.c.v.) and NT1 (10.0 and 20.0 mg/kg, i.p.) did not produce catalepsy. A much lower dose of neurotensin (0.03 microgram, i.c.v.) significantly reduced amphetamine- and phencyclidine-stimulated locomotor activity; NT1 also diminished amphetamine- and phencyclidine-stimulated locomotion with ED50 values of 0.3 and 0.4 mg/kg, i.p., respectively. Neurotensin (0.01-0.3 microgram, i.c.v.) and NT1 (0.1-1.0 mg/kg, s.c.) also produced dose dependent analgesia in the paw pressure test and decreased body temperature; these effects were insensitive to pretreatment with naloxone (10.0 mg/kg, i.p.). Together, the results support the hypothesis that neurotensin agonists have antipsychotic and analgesic activity. Moreover, the data suggest that such compounds may not produce extrapyramidal side effects. PMID- 9396067 TI - About-half-weekly (circasemiseptan) component of the endothelin-1 (ET-1) chronome and vascular disease risk. AB - Plasma ET-1 was measured around the clock on different calendar dates in healthy subjects and in subjects with diabetes and/or with high blood pressure and/or a history of vascular complications (HVDR). A transverse approach, with each subject contributing a single 24-h mean, assessed any about-weekly or half-weekly variation in ET-1. A circasemiseptan component resolved by single cosinor for nondiabetic (but not for diabetic) HVDR subjects (p = 0.010) differs in its timing of overall high values (p < 0.050) from that found in healthy subjects (p = 0.006). The results are aligned with circasemiseptan patterns in other circulatory variables and morbidity/mortality statistics. PMID- 9396066 TI - Synthesis of recombinant atrial natriuretic peptide (rANP) using hybrid fusion protein-phage fr coat/ANP (CP/ANP). AB - Recombinant atrial natriuretic peptide (rANP) was expressed in and isolated from E. coli. rANP was purified using HPLC. Amino acid analysis, partial sequencing, and molecular mass were determined. Fused protein was used to rise polyclonal antibodies and to develop of immunoenzymatic assays of rANP and CP/ANP. Experiments were designed to study rANP effects on isolated rabbit aortic strips and to examine hypotensive, diuretic, and natriuretic activity, as well as renal creatinine clearance, in an in vivo rat model. Identity of recombinant and commercial ANP has been confirmed. Physiological activity of CP/ANP has allowed the investigators to predict the conformation of CP/ANP, pro-ANP processing, and the method by which fusion protein interacts with ANP receptors. PMID- 9396068 TI - Expression of corticotropin-releasing factor (CRF) in peritoneum and pericardium in the rat embryo by in situ hybridization histochemistry. AB - Corticotropin releasing factor (CRF) mRNA expression in the thorax and abdomen was studied by in situ hybridization in rat tissue sections obtained from embryonic day 12 (E12) to E16. On E12 no signal was observed in any region. On E13 the first signals were detected particularly in serous membranes such as pericardium and peritoneum. By E14, the signal was strong in these membranes. On E15 the signal markedly diminished, and on E16 no signal was evident in serous membranes. This stage-specific expression of CRF mRNA in developing pericardium and peritoneum suggests that this peptide acts in pericardial and peritoneal development. PMID- 9396069 TI - Comparison of the antisecretory effect of endogenous forms of peptide YY on fed and fasted rat jejunum. AB - It is intriguing that the antisecretory peptide YY is present in plasma in two forms: PYY1-36 and PYY3-36. PYY3-36 has been found in human and rabbit blood within 30 min of the beginning of the meal, when the peak of water and electrolyte secretion occurs in the duodeno-jejunum. The aim of this study was therefore to compare the antisecretory effect of PYY1-36 and PYY3-36 in fed and fasted rat jejunum. The variations in electrolyte secretion were assessed by measuring the variations in short-circuit current (delta Isc) and transepithelial isotopic chloride fluxes in jejunal mucosa isolated from fed and fasted animal, and mounted in Ussing Chambers. In fasted animals, 2 x 10(-7) M PYY3-36 induced a reduction in Isc of -0.50 +/- 0.01 microEq/hr.cm2, which was not statistically different from that induced by 2 x 10(-7) M PYY1-36 (-0.60 +/- 0.01 microEq/h cm2). In contrast, in fed animals, 2 x 10(-7) M PYY3-36 did not trigger a significant response on Isc and net chloride flux, while the response to PYY1-36 was present but blunted. The absence of response was probably not related to the presence of secretory peptides because PYY3-36 was still able to induce a reduction in Isc after stimulation by a series of 10 different secretory peptides. After 10(-8) M PYY3-36 addition to an epithelium from the fasted animal, response to 10(-7) M PYY3-36 was blunted for 30 min and returned to control value after 60 min. Plasma concentration of PYY was higher in the fed rats compared to fasted (213.78 +/- 38 vs. 53.62 +/- 11.47 pg/ml p < 0.01). After incubation of crypt cells with or without 0.1 microM of unlabeled PYY for 60 min, Scatchard analysis of equilibrium binding data show that binding capacity (Bmax) of receptors was reduced when crypt cells were previously incubated with unlabeled PYY without significant modification of dissociation constants. Bmax were 183 +/- 27 in control vs. 56 +/- 11 fmol/mg protein. These results confirm the antisecretory activity of PYY1-36 in the jejunum of fasted and fed rats. They further indicate that PYY3-36 displays similar activity to PYY1-36 in fasted animals, but lack of activity in fed animals. These results suggest that the two circulating forms of PYY act as antisecretory peptides by two different mechanisms, implying a C-terminal specificity. PMID- 9396070 TI - Selective, physiological transport of insulin across the blood-brain barrier: novel demonstration by species-specific radioimmunoassays. AB - Insulin in blood is thought to cross the blood-brain barrier (BBB) to act within the brain to help control appetite. We examined the ability of blood-borne insulin to cross the BBB. Human insulin was infused for 48 h subcutaneously at several doses into mice and the amount of human and murine insulin in serum and brain measured with species-specific radioimmunoassays. For the exogenous human insulin, both brain and blood concentrations increased with increasing doses of infused insulin, whereas for the endogenous murine insulin, brain and blood concentrations decreased. Since the mouse cannot make human insulin, blood was the only source for the human insulin in brain, demonstrating that insulin does indeed cross the BBB. The relationship between the concentrations of human insulin in brain and blood was nonlinear, showing that passage is by a saturable mechanism. Partial saturation of the transporter occurred at euglycemic concentrations of serum insulin. Thus, insulin enters the brain by a saturable transport system that is operational primarily at physiological levels of serum insulin. PMID- 9396071 TI - Leptin stimulates insulin secretion and synthesis in HIT-T 15 cells. AB - Leptin, an ob gene product, corrects hyperinsulinemia in ob/ob mice. The leptin receptor may exist in pancreatic islets. The present studies were undertaken to determine the direct effect of 1-100 ng/ml recombinant leptin on insulin secretion and synthesis in HIT-T 15 cells by using static culture system. The addition of recombinant leptin significantly increased insulin secretion for 20 min at the highest concentration (100 ng/ml). The addition of recombinant leptin dose-dependently increased insulin secretion for 24 h in the 7 mM glucose containing F-12 K medium. The incubation with recombinant leptin for 24 h increased preproinsulin mRNA expression, assessed with reverse transcription polymerase chain reaction (RT-PCR) method. It was furthermore demonstrated that HIT-T 15 cells possessed the specific binding site for [125I]-labeled leptin. The present study demonstrated the existence of the leptin-specific binding sites that mediate its stimulatory effect on insulin secretion and synthesis in HIT-T 15 cells. PMID- 9396072 TI - Leptin: a potent inhibitor of insulin secretion. AB - The hormone leptin is expressed and secreted by the adipose tissue and impacts on the central nervous system. Leptin is involved in the regulation of energy balance, satiety, and body composition. The lack of active leptin results in obesity, high food intake, hyperglycemia, and hyperinsulinemia. We present data supporting effects of leptin on the endocrine pancreas. We found the leptin receptor to be expressed in insulin- and glucagon-secretin cells derived from mouse, hamster, and rat pancreas. In the isolated perfused rat pancreas leptin is a potent inhibitor of basal and glucose-induced insulin secretion, especially during the first phase of the insulin response. At isolated mouse islets and insulin-secreting INS-1 cells leptin reduced promptly and persistently the intracellular Ca2+ levels. Cytoplasmic Ca2+ oscillation amplitude was decreased and the oscillation frequency increased. These findings suggest functional active receptors for leptin on insulin-secreting B-cells. Therefore, leptin is a metabolic hormone and not only a signal to the brain indicating filled fat stores. Our data suggest that leptin is also a signal back to the endocrine pancreas that no more insulin is required to replenish fat stores. Thus, an "adipo-insular axis" operating with two arms exists: insulin and glucagon are signals to the adipocyte. This releases leptin, which could be the mediator of the respective feedback to the pancreas. A defective leptin suppression of insulin secretion could contribute to hyperinsulinemia and disturbances of glucose metabolism. PMID- 9396073 TI - Synergy between leptin and cholecystokinin (CCK) to control daily caloric intake. AB - Both cholecystokinin (CCK), a short-term meal-related satiety signal, and the ob protein leptin, a postulated long-term adiposity hormone, are thought to be important signals in the multiple interacting systems that control appetite and adiposity. We hypothesized that these hormones may synergistically interact to suppress feeding. Following IP administration of leptin (two doses of 50 micrograms each) and CCK (2, 4, 8, or 16 micrograms) total daily caloric intake was significantly reduced by leptin and CCK compared to leptin alone. These results support the hypothesis that CCK and leptin may synergistically interact to control long-term feeding. PMID- 9396075 TI - Effects of acute and chronic administration of L-arginine on the antinociceptive action of morphine-6-beta-D-glucuronide. AB - Chronic administration of L-arginine (200 mg/kg i.p.) but not of D-arginine (200 mg/kg i.p.) twice a day for 4 days decreased the antinociceptive response to subcutaneously administered morphine-6-beta-D-glucuronide (M6G), a potent metabolite of morphine, in male Swiss-Webster mice as measured by the tail-flick test. However, the antinociceptive response to intracerebroventricularly administered M6G was unaffected by chronic treatment with L-arginine. The decreased antinociceptive response to M6G (s.c.) was reversed by concurrent administration of NG-nitro-L-arginine (5 mg/kg i.p.), an inhibitor of nitric oxide synthase. Acute administration of L-arginine (200 mg/kg i.p.) had no effect on M6G-induced antinociception, but higher doses (400 and 800 mg/kg i.p.) decreased it. Since the antinociceptive response to centrally administered M6G was unaffected by chronic L-arginine treatment, the decreased antinociceptive response of peripherally administered M6G may be related to a decrease of M6G entry into brain structures responsible for antinociceptive action. PMID- 9396074 TI - Brain melanocortin receptors: from cloning to function. AB - The cloning of brain melanocortin (MC) receptors, the mapping of their expression pattern and the identification of MC receptor selective ligands have opened a new avenue towards elucidating the role of the melanocortin system in the brain. MC receptors have now been implicated in melanocortin-induced grooming behavior in rats, in the melanocortin-induced lowering of blood pressure and in the control of weight homeostasis. Functional opioid antagonism and the anti-pyretic and anti inflammatory effects of melanocortins are probably also mediated via MC receptors. However, the effects of melanocortins on avoidance behavior and the effect of gamma 2-MSH on increasing blood pressure are not mediated via one of the cloned brain MC receptors. The structure of brain MC receptors, their expression pattern, the MC receptor selective ligands and the function of MC receptors are briefly reviewed. PMID- 9396076 TI - Anti-inflammatory action of methimazole. AB - The anti-inflammatory activity of 1-methylimidazole-2-thiol (methimazole), the most widely used antithyroid drug, was investigated. Methimazole had a marked inhibitory action on prostaglandin H synthase (IC50 = 10 mumol/l), inhibiting the peroxidase (IC50 = 330 mumol/l), although the cyclo-oxygenase was slightly activated. Methimazole was less potent than indometacin (IC50 = 1.7 mumol/l) on prostaglandin H synthase, but was more potent than acetylsalicylic acid (IC50 = 160 mumol/l). Methimazole has been found to trap superoxide (O2.-) radicals and to decrease the level of blood prostaglandin E2. PMID- 9396078 TI - Effect of mucosal resection on detrusor function in the disused rabbit bladder. AB - A new rabbit model of disused bladder was developed. Disused bladders were created by sagittal splitting of the bladders along the midline and closure of one side (reservoir side), leaving the remaining side unclosed (disused). The mucosa was dissected from some of the disused sides. Tissue elasticity and the weight of the detrusor muscle were decreased by bladder disuse. The responsiveness to field stimulation, bethanechol, ATP, KCl, or isoproterenol, was decreased by disuse. Tissue elasticity and responsiveness to stimuli were decreased to a lesser extent in disused tissue with intact mucosa. Results suggest that the bladder mucosa helps to preserve the function of the disused bladder. PMID- 9396077 TI - Lysine14galanin(1-15)-NH2: a partial agonist at galanin receptors in rat isolated gastric fundus. AB - The study was undertaken to characterize the effects of the porcine galanin [pGal(1-29)-NH2] analogue [Lys14]pGal(1-15)-NH2 on rat gastric fundus. [Lys14]pGal(1-15)-NH2 is a less potent contractile agent than pGal(1-29)-NH2 (EC50 74.1 vs. 43.7 nmol/l, respectively) and shows a significantly lower maximal response than pGal(1-29)-NH2. Concentration-contraction curves were constructed for pGal(1-29)-NH2 alone (control) and pGal(1-29)-NH2 in the presence of 10, 100, and 1,000 nmol/l of [Lys14]pGal(1-15)-NH2. [Lys14]pGal(1-15)-NH2 shifted the concentration-contraction curves of pGal(1-29)-NH2 significantly to the right, whereas their linear portions remained parallel to that for the pGal(1-29)-NH2 control. [Lys14]pGal(1-15)-NH2 markedly increased the EC50 of the respective pGal(1-29)-NH2 concentration-contraction curves. It did not substantially change the maximal response of the muscles to pGal(1-29)-NH2 and the form of the respective concentration-contraction curves. Schild's plot gave a straight line with a slope of 0.84. The pA2 value for [Lys14]pGal(1-15)-NH2 was 8.23. [Lys14]pGal(1-15)-NH2 seems to be a partial Gal receptor agonist. Since the lack of specific Gal receptor antagonists in the gastrointestinal tract makes a precise characterization of its role as a motility modulator difficult, the position 14 in the pGal(1-29)-NH2 molecule looks as an attractive target in the search of a pure Gal receptor antagonist in the smooth muscles of the gut. PMID- 9396079 TI - Effects of KW-3902, a selective and potent adenosine A1 receptor antagonist, on renal hemodynamics and urine formation in anesthetized dogs. AB - The purpose of the present investigation was to examine the effects of KW-3902 [8 (noradamantan-3-yl)-1,3-dipropyl-xanthine], a selective and potent adenosine A1 receptor antagonist, in order to clarify the role of adenosine in the control of renal hemodynamics and urine formation in anesthetized dogs. KW-3902 was directly infused into the renal artery to eliminate the systemic effects of the drug. KW 3902 (10 micrograms/kg/min) almost completely inhibited the renal vasoconstriction induced by adenosine via A1 receptors. Intrarenal infusion of KW 3902 did not affect mean arterial pressure, renal blood flow, creatinine clearance, or arterial plasma renin activity, but drastically increased urine flow, urinary excretion of sodium, and osmolar clearance. Inhibition of the renin angiotensin system using CV-11974 [2-ethoxy-1-((2'-(1-H-tetrazole-5-yl)biphenyl-4 yl) methyl)-1-H-benzimidazole-7-carboxylic acid], a selective AT1 antagonist, did not affect the diuretic action of KW-3902. These data suggest that endogenous adenosine does not play a significant role in the control of renal hemodynamics in whole kidney, but that it plays an important role in preserving body fluid via the A1 receptor independent of the renin-angiotensin system in anesthetized dogs. PMID- 9396080 TI - Nitric oxide reduces myocardial contractility in isoproterenol-stimulated rat hearts by a mechanism independent of cyclic GMP or cyclic AMP. AB - Nitric oxide has been shown to decrease myocardial contractility and O2 consumption. This study was designed to evaluate the hypothesis that nitric oxide mediated increases in cyclic GMP require elevated cyclic AMP to produce cardiac depression. Using isolated, Langendorff-perfused rat hearts, we determined the effects of intracoronary nitroprusside (NP, 1 and 10 mM) in the absence and presence of isoproterenol (ISO, 10(-8) M) on cardiac function, O2 consumption, cyclic GMP and cyclic AMP. ISO, with and without NP, increased cyclic AMP (from 287 +/- 21 to 477 +/- 33 pmol/g) without altering cyclic GMP. Left-ventricular pressure increased from 97 +/- 12 to 178 +/- 9 mm Hg and dP/dtmax from 1,786 +/- 275 to 4,049 +/- 354 mm Hg/s. NP increased cyclic GMP (from 4 to 30 pmol/g) in both the absence and presence of ISO, but NP did not alter cyclic AMP. Without ISO, NP insignificantly altered left-ventricular pressure; however, in the presence of ISO, NP significantly decreased left-ventricular pressure by -25 +/- 4 mm Hg and decreased dP/dtmax by -619 +/- 142 mm Hg/s. Isoproterenol increased O2 consumption, but the changes with NP were not significant. When this study was repeated in the presence of LY83583, a guanylate cyclase inhibitor, NP still produced cardiac depression in the presence of ISO. Therefore, cardiodepressant effects of NP were only observed against a background of inotropic stimulation with ISO. However, effects of NP on contractility were unrelated to increases in cyclic GMP or cyclic GMP-induced changes in cyclic AMP. PMID- 9396081 TI - Moxonidine-induced inhibition of norepinephrine release in monkey and rabbit ciliary bodies: role of cGMP. AB - This study was designed to determine whether in isolated rabbits iris-ciliary bodies and monkey ciliary bodies, cGMP plays a role in the action of moxonidine, an alpha 2- and imidazoline (I1) receptor agonist. In field-stimulated rabbit iris-ciliary bodies, dose-related inhibition of norepinephrine release was induced by 8-Br-cGMP, moxonidine or sodium nitroprusside; 8-Br-cGMP in combination with moxonidine did not enhance inhibition of norepinephrine release. Sodium nitroprusside at intermediate and high concentrations stimulated cGMP production in rabbit iris-ciliary bodies, whereas moxonidine stimulated cGMP production modestly only at a high concentration. When iris-ciliary bodies were pretreated with a low concentration of moxonidine, sodium nitroprusside stimulated cGMP production was enhanced from 1.6 to 2.2 pmol/mg protein. In field stimulated monkey ciliary bodies, both sodium nitroprusside and moxonidine inhibited norepinephrine release. Pretreatment of electrically stimulated monkey ciliary bodies with sodium nitroprusside enhanced the suppressive effect of moxonidine on norepinephrine release. In monkey ciliary bodies, moxonidine raised cGMP production more than sodium nitroprusside did, but there was no synergism in cGMP production by combined treatment with moxonidine and sodium nitroprusside. These results suggest that cGMP could play a role in the ocular action(s) of moxonidine in ciliary bodies; however, involvement of cGMP in the action of moxonidine in monkey ciliary bodies seems to be more pronounced than in rabbit iris-ciliary bodies. PMID- 9396082 TI - Intestinal fate of 5-aminosalicylic acid: regional and systemic kinetic studies in relation to inflammatory bowel disease. PMID- 9396083 TI - Effects of 3-deazaadenosine on apoptosis-related gene transcripts in HL-60 cells. AB - The effect of the transmethylation inhibitor 3-deazaadenosine on transcription levels of genes associated with apoptosis was investigated in HL-60 cells. After incubation of HL-60 cells with 100 microM 3-deazaadenosine for 45 min., a schedule known to perturb transmethylation metabolites and initiate apoptosis in these cells, a 50% decrease in c-myc and a 50% increase in bcl-2 RNA steady-state levels compared to control cells were observed. Transcription levels of c-myc continued to decrease after extended exposure to 3-deazaadenosine, while bcl-2 mRNA levels dropped to 25% and 30% below those in control cells after 1.5 hr and 3 hr, respectively. The expression levels of the bcl-2 related bax gene, showed a similar pattern as bcl-2; a 60% increase was initially measured, but after 1.5 and 3 hr, bax transcripts were 80% and 70% respectively, of those found in untreated cells. Another bcl-2 related gene, bcl-x, was previously reported to generate two transcripts in human cells. The long variant bcl-x1 acts as bcl-2, while the short form bcl-xs induces apoptosis. We were unable to detect bcl-xs transcripts in untreated and 3-deazaadenosine treated cells by the highly sensitive reverse transcriptase polymerase chain reaction method. This suggests that this gene product may not be involved in 3-deazaadenosine induced apoptosis in HL-60 cells. Bcl-x1 mRNA levels, however, slowly decreased with about 50% after 1.5 or 3 hr 3-deazaadenosine treatment. It is concluded that 3 deazaadenosine initiated apoptosis affects c-myc, bcl-2, bax and bcl-x1 mRNA levels. PMID- 9396084 TI - Analysis of native human plasma proteins and haemoglobin for the presence of bityrosine by high-performance liquid chromatography. AB - Generation of reactive oxygen species in vivo results in oxidative-damage to cellular components, including proteins. Due to the relatively long half-lives of several blood proteins the cumulative formation of oxidatively damaged proteins might serve as a biomarker for reactive oxygen species formation. The most prominent sources of reactive oxygen species in vivo are site-specific metal ion catalyzed reactions of the Fenton and Haber-Weiss types and the H2O2/peroxidase system. In vitro oxidation of L-tyrosine using a peroxidase or Cu++/H2O2 system gives rise to the formation of a highly fluorescent substance, bityrosine. High performance liquid chromatography (HPLC) analysis of acid hydrolyzed serum albumin after oxidation with peroxidase/H2O2 or with Cu++/H2O2 showed that bityrosine had been formed whereas oxidation of this protein with Fe(III)/ascorbate did not result in the formation of bityrosine. Bityrosine could not be detected in human plasma proteins or haemoglobin with the detection limit of one pmol per mg protein. PMID- 9396085 TI - Involvement of cholinergic and opioid receptor mechanisms in nicotine-induced antinociception. AB - In this work we have studied the influences of nicotinic agents on the antinociception of morphine in formalin test. Nicotine (0.001-0.1 mg/kg) induced antinociception in mice in a dose-dependent manner in the early phase of formalin test, and also potentiated the morphine effect. The nicotinic receptor antagonist, mecamylamine (0.5 mg/kg), but not hexamethonium decreased the antinociception induced by nicotine (0.1 mg/kg) in both phases. The muscarinic receptor antagonist atropine (5 and 10 mg/kg) also decreased the response of nicotine. Mecamylamine, hexamethonium or atropine did not alter morphine antinociceptive response, while naloxone decreased responses induced by nicotine or morphine. The antagonists by themselves did not elicit any response in formalin test, however, high does of mecamylamine tend to increase pain response. It is concluded that central cholinergic and opioid receptor mechanisms may be involved in nicotine-induced antinociception. PMID- 9396086 TI - Forskolin potentiates isoprenaline-induced glycerol output and local blood flow in human adipose tissue in vivo. AB - The synergistic action of forskolin on beta-adrenoceptor-mediated glycerol output and changes in local blood flow were investigated in situ, in human adipose tissue of healthy subjects, by the use of microdialysis. The addition of isoprenaline 0.1-1.0 microM or forskolin 10-100 microM to the perfusion solvent caused a concentration-dependent, marked and sustained increase in the levels of glycerol in the dialysate (lipolysis index) as compared to the solvent alone. On a molar basis, isoprenaline was almost one thousand times more potent than forskolin. Isoprenaline caused a rapid and concentration-dependent decrease in the ethanol clearance ratio (index of local blood flow, i.e. a decrease in ethanol ratio implies an increase in blood flow). Forskolin had no effect on the ethanol ratio at either 1.0 microM or 10 microM, while forskolin at 100 microM induced a significant decrease in the ethanol ratio. When adipose tissue was pre treated with forskolin, the subsequent addition of isoprenaline to the microdialysate resulted in a significantly higher glycerol output and a significantly more prominent decrease in the ethanol ratio than with isoprenaline alone. In conclusion, the data demonstrate that forskolin and the (beta adrenoceptor-agonist both stimulate lipolysis and local blood flow in human adipose tissue in vivo. Furthermore, forskolin, at concentrations that are ineffective alone, potentiates the actions of isoprenaline on lipolysis and blood flow. PMID- 9396087 TI - Actions of spermicidal and virucidal agents on electrogenic ion transfer across human vaginal epithelium in vitro. AB - Human ectocervical tissue was removed at operation over the menstrual cycle mounted as a sheet in vitro in an Ussing-style chamber and incubated in bicarbonate saline. The net electrogenic ion transport was measured as the short circuit current (Isc in muamps/cm2) and was characterised as mainly (60-86%) an amiloride-sensitive electrogenic Na+ transport (lumen to serosa). Serosal application of amiloride had no effect. Serosal application of ouabain, a selective Na(+)-pump inhibitor, reduced the Isc to near zero but neither theophylline (10 mM) nor furosemide (1 mM) had any action. The data are compatible with a model ectocervical vaginal cell having an amiloride-sensitive Na+ entry mechanism at the lumenal membrane and a Na(+)-pump at the basolateral membrane removing the ion from the cell. The effects of the putative virucides, chlorhexidine and benzalkonium chloride, were tested on the preparation. Mucosally added chlorhexidine (2 mg/ml) had no effect on the Isc or tissue resistance but benzalkonium chloride, at concentrations between 0.06-1.2%, caused a rapid fall in the Isc. At the highest concentration this was only partly reversible even after two washes with fresh buffer. At the lowest concentration (0.03%) benzalkonium chloride sometimes caused an initial increase in the Isc which then fell to zero. In all the tissues even after the Isc was reduced to near zero, nigrosin left in contact with the tissue for 5 min. did not enter and stain the cells, indicating the detergent had a selective membrane action rather than causing a non-specific increase in permeability. The preparation allows objective measurements to be made of the initial acute membrane actions of putative spermicides and virucides on human vaginal ectocervical epithelial cells and offers a new approach of assessing their pharmacological/toxicological actions. PMID- 9396088 TI - Comparison of key steps in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) neurotoxicity in rodents. AB - Three steps in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) neurotoxicity were compared with the neurodegenerative effects of the toxin in mice and rats. Firstly, we compared the neurotoxicity of MPTP, mediated by monoamine oxidase (MAO)-B, to that of 1-methyl-4-(2'-methylphenyl)-1,2,3,6-tetrahydropyridine (2' CH3-MPTP), an analogue oxidized by MAO-A and MAO-B. Both toxins caused degeneration of dopamine terminals in mice but not in rats. In NMRI mice noradrenaline terminals were also affected by both toxins. Pretreatment with deprenyl to prevent MAO-B-mediated oxidation in the capillary endothelium enhanced dopamine toxicity to 2'-CH3-MPTP in nucleus accumbens but no potentiation was seen in striatum and the olfactory tubercle. Secondly, synaptosomal uptake of the 1-methyl-4-phenylpyridinium ion (MPP+) was studied. Uptake in rats was not significantly different from that in the two mice strains. Thirdly, no significant differences were found in MPP(+)-induced lactate production in striatal slices or synaptosomes. We conclude that the lack of effect of MPTP in rats is not due to mechanisms specific for MPTP but probably to the ability of rat catecholamine neurons to cope with, and survive, impaired energy metabolism. PMID- 9396089 TI - Evaluation of three novel cholecystokinin-B/gastrin receptor antagonists: a study of their effects on rat stomach enterochromaffin-like cell activity. AB - Gastrin stimulates rat stomach enterochromaffin-like (ECL) cells via activation of cholecystokinin-B/gastrin receptors. The stimulation is manifested in the activation of the histamine-forming enzyme histidine decarboxylase and in the secretion of histamine and pancreastatin, a chromogranin A-derived peptide. We have examined the short-term effects of three novel cholecystokinin-B/gastrin receptor antagonists (YF476, JB93182 and AG041R) on the ECL cells in intact fasted rats. The drugs and/or gastrin were infused intravenously for 3 hr and the oxyntic mucosal histidine decarboxylase activity and the serum pancreastatin concentration were measured. We also studied the effects of the three drugs on gastric emptying in mice, a cholecystokinin-A receptor-mediated response. YF476, JB93182 and AG041R antagonized the gastrin-evoked histidine decarboxylase activation in a dose-dependent manner. YF476, JB93182 and AG041R induced maximal inhibition at 0.03, 0.1 and 0.1 mumol kg-1 hr-1, respectively; the corresponding ID50 values were 0.002, 0.008, and 0.01 mumol kg-1 hr-1. YF476 was selected for further analysis. It produced a rightward shift of the gastrin dose-response curve, consistent with competitive inhibition. Moreover, it antagonized the omeprazole-evoked histidine decarboxylase activation and the gastrin- and omeprazole-induced rise in the circulating pancreastatin concentration. None of the three drugs tested inhibited gastric emptying or prevented the cholecystokinin-8s-induced inhibition of gastric emptying at the doses tested. The results show that YF476, JB93182 and AG041R are potent and selective cholecystokinin-B/ gastrin receptor antagonists, and that YF476 is 4-5 times more potent than JB93182 and AG041R. PMID- 9396090 TI - The effects of glyceryl trinitrate and nicotinic acid ointments during cold exposure. AB - The aim of the present experimental study was to examine the effects of local application of glyceryl trinitrate and nicotinic acid on the cold-provoked haemodynamic responses, pain and hand dexterity. Ten young healthy volunteers participated in this randomized, cross-over study with three phases at least two days apart. Five cm of 2% glyceryl trinitrate ointment, 10% nicotinic acid ointment or placebo ointment was applied on the back of each subject's both hands 15 min. before the 7 min. cold exposure. Blood pressure and heart rate were measured prior to, during and after the cold exposure. In addition, the effect of cold on hand dexterity was evaluated by the Purdue pegboard test and the subjects assessed the pain in their hands during the cold exposure. Pretreatment with glyceryl trinitrate ointment counteracted the cold-induced haemodynamic response, as evidenced by a significantly (P < 0.05) smaller mean increase in the systolic blood pressure from the baseline compared with placebo. In contrast, the cold induced increase in the systolic blood pressure observed after pretreatment with nicotinic acid ointment did not differ from placebo. Both glyceryl trinitrate and nicotinic acid alleviated the cold-induced pain, but neither of them prevented the deterioration of hand dexterity. In conclusion, the haemodynamic response provoked by a brief cold exposure could to some extent be counteracted by pretreatment with glyceryl trinitrate ointment, but not with nicotinic acid ointment, compared with placebo. PMID- 9396091 TI - Combined oral treatment with racemic and meso-2,3-dimercaptosuccinic acid for removal of mercury in rats. AB - Racemic dimercaptosuccinic acid (DMSA) was found more efficient than the meso isoform in enhancing the removal of mercury in rats. However, racemic-DMSA has recently been found more toxic. The efficiency of combined oral treatment with the two isoforms of DMSA for removal of mercury has now been evaluated. Female albino rats were treated orally for four days with meso- (M) and/or racemic- (R) DMSA (1 mmol/kg each), five days after a single intraperitoneal administration of 203Hg with 0.5 mg HgCl2/kg. The animals were divided into six groups according to the number of treatments with each isomer: control (untreated), 4M, 1R + 3M, 2R + 2M, 3R + 1M, and 4R. Whole body, kidney, liver and brain mercury contents were measured nine days after 203Hg administration. In all treated groups retention in the whole body and kidneys was greatly reduced. The groups treated with racemic DMSA, regardless of the number of doses, showed a greater removal of mercury than the group treated with meso-DMSA alone (4M). All treatments were less efficient in reducing liver retention, and the brain retention was not affected. It was concluded that even a single application of the more toxic racemic-DMSA during a four-day oral treatment regimen is sufficient to improve the removal by meso-DMSA of mercury from rats. PMID- 9396092 TI - Vancomycin removal by plasmapheresis. PMID- 9396093 TI - Reducing tobacco harms among older adults: a critical agenda for tobacco control. PMID- 9396094 TI - Back to basics: getting smoke-free workplaces back on track. PMID- 9396095 TI - The Farmington consensus statement on editorial guidelines for addiction journals. PMID- 9396097 TI - Turkey: Camel gets through again. PMID- 9396096 TI - The pack as advertisement. PMID- 9396098 TI - India: tobacco toothpaste squeezed out. PMID- 9396099 TI - Blaming the children. PMID- 9396100 TI - Cigarette smoking and smoking cessation among older adults: United States, 1965 94. AB - OBJECTIVE: To characterise patterns of cigarette smoking and smoking cessation among older adults in the United States. DESIGN: Data from the National Health Interview Surveys (NHIS) 1965-94 were analysed. The NHIS is a cross-sectional survey using a representative national sample. SETTING: In most cases interviews were conducted in the home; telephone interviews were conducted when respondents could not be interviewed in person. PARTICIPANTS: Participants were from a representative sample of the American civilian, non-institutionalised population aged 18 and older. Sample sizes for the years analysed ranged from n = 19,738 to n = 138,988 overall, and n = 3806 to n = 12,491 for those aged 65 years and older. MAIN OUTCOME MEASURES: Using the NHIS data from 1965-94, trends in current smoking and the prevalence of smoking cessation by demographic characteristics among older adults (65 years and older) were assessed and compared with trends among younger adults. A logistic regression analysis was conducted to determine the demographic characteristics of former smokers compared with current smokers among those aged 65 and older. RESULTS: The prevalence of current smoking among 65 year olds and older declined from 1965 to 1994 (17.9% to 12.0%). Although smoking prevalence was lower among older adults than younger adults (aged 18-64), the rate of decline in smoking was slower among older adults. Among older adults, the prevalence of cessation rose with increasing educational attainment, and was consistently higher for men than for women and for whites compared with blacks. After adjustment for demographic factors among older adults who had ever smoked, increasing age and educational attainment were strongly related to the likelihood of being a former smoker. Although there were no racial differences among women, older white (OR = 2.6) and Hispanic (OR = 3.67) men were significantly more likely to be former smokers than older black men. Also, the gender difference in smoking cessation was noted only for whites. CONCLUSIONS: Given the projected increase in the elderly population, the medical and economic consequences of smoking will become a greater burden in the next decades. Therefore, focusing attention on cessation among the elderly is an immediate and urgent priority for public health professionals and clinicians. PMID- 9396101 TI - Predictors of smoking cessation among elderly smokers treated for nicotine dependence. AB - OBJECTIVE: To examine outcomes and predictors of smoking cessation among elderly patients treated for nicotine dependence. DESIGN: Retrospective analysis of patients aged 65-82 who received a nicotine dependence consultation at the Mayo Medical Center between 1 April 1988 and 30 May 1992. Patients were contacted by telephone by a trained interviewer six months after the consultation and were sent a follow-up survey in August 1993. SETTING: Mayo Medical Center, Rochester, Minnesota, United States. SUBJECTS: A total of 613 patients (310 men, 303 women) with a mean age of 69.0 (SD 3.5) years were seen during the study period. MAIN OUTCOME MEASURES: Point prevalence self-reported smoking status. Patients were considered abstinent if they self-reported not smoking (not even a puff) during the seven days before contact. RESULTS: At six-month follow up, 24.8% of the 613 patients reported abstinence from smoking. On multivariate analysis, smoking abstinence was more likely if patients were hospitalised at the time of the consultation, married to a non-smoking spouse, very motivated to stop smoking, and reported their longest time of previous abstinence to be less than a day or more than a month. The response rate to the mailed follow-up survey was 69.9% (429 of 613). The mean duration of follow up was 40.0 +/- 13.2 months following the consultation. Of the 429 patients, 103 (24.0%) reported abstinence from smoking and 326 (76.0%) were smoking at six-month follow up. Patients who reported abstinence at six months had a higher cessation rate at the last follow up (76.0%) compared with patients who were smoking at six-month follow up (33.0%, P < 0.001). For patients who were not smoking at six months, no factors were found to significantly predict abstinence at last follow up. For patients who were smoking at six months, factors associated with smoking cessation at last follow up were: more than a year as the longest time off cigarettes before the consultation; counsellor rating of less severe nicotine dependence; and older age at first regular smoking. CONCLUSIONS: Several predictors of smoking cessation were identified in this study which may be useful for tailoring smoking interventions for the elderly. PMID- 9396102 TI - Self-help interventions for older smokers. AB - OBJECTIVE: To evaluate the relative effectiveness of two self-help smoking interventions as adjuncts to a self-help manual and telephone support service (hotline) for older smokers. DESIGN: Subjects were stratified on baseline variables and randomised to one of two treatment conditions in a methods development study. SUBJECTS: 177 community-dwelling smokers aged 60 years and older. INTERVENTIONS: All subjects received a self-help manual and access to a smokers' telephone hotline. Subjects also received either mailings (Letters condition) or counselling telephone calls (Proactive condition) at four and eight weeks after enrollment. MAIN OUTCOME MEASURES: Use of the hotline and prevalence of abstinence lasting at least 48 hours (verified by a "significant other") were assessed at three and six months for the full sample. Seven-day abstinence was calculated for comparison with previous research. A subsample of 91 subjects was followed up at 12 months. RESULTS: Overall abstinence rates for the two conditions were in the range of typical self-help interventions. Men were more likely to be abstinent than women at follow up at three and six months. A significant gender x treatment interaction was found, with abstinence rates higher for men in the Letters condition, and women in the Proactive condition. Hotline use was high, with nearly half of subjects calling by 12 months. CONCLUSION: Both interventions appear promising for older smokers, but may be differentially effective for men and women. Older smokers will use a hotline; whether Letters and Proactive interventions can improve on manual and hotline effectiveness rates alone is being tested in a subsequent controlled trial. PMID- 9396103 TI - Non-smoking policies, tobacco education, and smoking cessation programmes in facilities serving the elderly in Michigan, United States. AB - OBJECTIVE: To determine the extent of and impetus for smoke-free policies in facilities serving Michigan's elderly, and the extent of tobacco education and smoking cessation programmes for elders and staff of these facilities. DESIGN: Telephone survey in February 1997 of three types of facilities serving Michigan's elderly population. SUBJECTS: Area Agencies on Aging (n = 12), Councils and Commissions on Aging (n = 31), and senior centres (n = 98) located in Michigan, USA. MAIN OUTCOME MEASURES: Prevalence of smoke-free policies, tobacco education, and smoking cessation programmes in facilities serving the elderly. RESULTS: 99% (95% confidence interval (CI) = 97% to 100%) of 141 facilities surveyed have an indoor smoke-free policy. Eighty-five per cent (95% CI = 79% to 91%) of these policies prohibit all smoking inside the facility. Forty-five per cent (95% CI = 37% to 54%) cited a law as requiring the smoke-free policy, whereas 38% (95% CI = 30% to 46%) indicated the policy was adopted voluntarily for health reasons. Forty-two per cent (95% CI = 34% to 50%) of the facilities provided some education on the dangers of tobacco, while 11% (95% CI = 6% to 16%) arranged smoking cessation programmes for staff or elders. CONCLUSIONS: In Michigan, a very high percentage of non-institutional facilities serving the elderly have smoke-free policies, which appear to increase participation at these facilities. Tobacco education programmes are provided in less than half the facilities, and very few arrange smoking cessation programmes for elders or staff. PMID- 9396104 TI - Workplace smoking policies in the United States: results from a national survey of more than 100,000 workers. AB - OBJECTIVE: To determine the prevalence of smoking policies in indoor work environments as reported by a nationally representative sample of workers in the United States. DESIGN: Cross-sectional survey of households within the United States. SETTING: All 50 state and the District of Columbia, 1992-93. PARTICIPANTS: Currently employed indoor workers 15 years of age and older who responded to the National Cancer Institute's Tobacco Use Supplement to the Current Population Survey (n = 100,561). MAIN OUTCOME MEASURES: The prevalence and restrictiveness of workplace smoking policies as reported by workers currently employed in indoor workplaces in the United States. RESULTS: Most of the indoor workers surveyed (81.6%) reported that their place of work had an official policy that addressed smoking in the workplace; 46.0% reported that their workplace policy did not permit smoking in either the public/common areas- for example, restrooms and cafeterias--or the work areas of the workplace. The reporting of these "smoke-free" policies varied significantly by gender, age, race/ethnicity, smoking status, and occupation of the worker. CONCLUSIONS: Although nearly half of all indoor workers in this survey reported that they had a smoke-free policy in their workplace, significant numbers of workers, especially those in blue-collar and service occupations, reported smoke-free rates well below the national average. If implemented, the US Occupational Safety and Health Administration's proposed regulation to require worksites to be smoke free has the potential to increase significantly the percentage of American workers covered by these policies and to eliminate most of the disparity currently found across occupational groups. PMID- 9396105 TI - A mass media programme to prevent smoking among adolescents: costs and cost effectiveness. AB - OBJECTIVE: To examine costs and cost-effectiveness ratios of a four-year mass media programme previously shown to prevent the onset of smoking among adolescents. DESIGN: A matched control design. SETTING: Two cities in Montana, one in New York and one in Vermont, USA. SUBJECTS: Students in grades 10-12 (ages 15-18). INTERVENTION: A four-year mass media campaign to prevent the onset of smoking. MAIN OUTCOME MEASURES: Cost per student potentially exposed to the mass media campaign; cost per student smoker potentially averted; and cost per life year gained. Cost estimates were also made for a similar campaign that would be broadcast nationally in the United States. RESULTS: In 1996 dollars, the cost of developing and broadcasting the mass media campaign was $759,436, and the cost per student potentially exposed to the campaign (n = 18,600) was $41. The cost per student smoker averted (n = 1023) was $754 (95% confidence interval (CI) = $531-$1296). The cost per life-year gained discounted at 3% over the life expectancy for young adult smokers was $696 (95% CI = $445-$1269). The estimated cost of developing and broadcasting a similar four-year mass media campaign in all 209 American media markets would be approximately $84.5 million, at a cost of $8 per student potentially exposed to a national campaign, $162 per student smoker averted, and $138 (95% CI = $88-$252) per life-year gained. CONCLUSION: Estimates of the cost-effectiveness ratios of this mass media campaign in preventing the onset of smoking showed it to be economically attractive and to compare favourably with other preventive and therapeutic strategies. PMID- 9396106 TI - Question 1 tobacco education expenditures in Massachusetts, USA. AB - BACKGROUND: In 1992, voters in Massachusetts (United States) approved Question 1, a state ballot initiative, which raised the state excise tax to provide funds for tobacco education. OBJECTIVE: To examine Question 1 expenditures for tobacco specific programmes in the 1994, 1995, 1996, and 1997 fiscal years. DESIGN: This study examined trends in Question 1 expenditures. Data were collected from the Massachusetts Department of Public Health and the Massachusetts Department of Revenue for the 1994, 1995, 1996, and 1997 fiscal years. MAIN OUTCOME MEASURES: The amount of spending on tobacco-specific programmes. RESULTS: Excluding the 1994 fiscal year because the state allocated 18 months of new revenues, from the 1995 fiscal year to the projected 1997 fiscal year, the state will have spent 22% of Question 1 funds for tobacco-specific programmes. Question 1 expenditures for tobacco-specific programmes have declined by 15%, whereas Question 1 expenditures for the other programmes decreased only 0.4%. CONCLUSIONS: The legislature has established a trend that has produced real reductions in Question 1 funding for tobacco education, which appears contrary to the mandate of the voters when they enacted Question 1 in 1992. These reductions undermine the effectiveness of tobacco-specific programmes that are an integral part of the Massachusetts Tobacco Control Programme. These results also highlight the fact that the initial compromises made after initiatives such as Question 1 are adopted have important long-term consequences for funding of tobacco control initiatives. PMID- 9396107 TI - Review of the evidence that pH is a determinant of nicotine dosage from oral use of smokeless tobacco. AB - OBJECTIVE: To determine whether manipulation of the pH of moist-snuff products by manufacturers could control the delivery of nicotine. DATA SOURCES: Medline database 1966-97 using the following subject headings and keywords: nicotine, absorption, mouth mucosa, skin, hydrogen-ion concentration, smokeless tobacco, biological transport, and membranes; computer database of the tobacco bibliography maintained by the US Centers for Disease Control and Prevention's Office on Smoking and Health; bibliographies of pertinent journal articles, books, and governmental reports; personal communications with experts in nicotine pharmacology and addiction; and Brown & Williamson Tobacco Corporation documents in the Tobacco Control Archives of the University of California, San Francisco. STUDY SELECTION: Included all relevant animal studies, in-vitro studies, nicotine replacement therapy trials, and human observational studies. DATA SYNTHESIS: We found that the effects of pH on drug absorption have been well established in animal models for nicotine and many other acidic or basic compounds. Increased alkalinity promotes the absorption of nicotine and increases its physiological effects. Human studies, which are more limited, confirm these processes. For example, nicotine absorption is directly related to the pH when nicotine is delivered in either tobacco smoke or nicotine polacrilex gum. CONCLUSIONS: Although other factors could influence the rate of nicotine absorption from oral tobacco, manipulating tobacco pH appears to be the primary means by which the speed of nicotine absorption is determined in moist-snuff products. PMID- 9396108 TI - Tobacco blindness. PMID- 9396109 TI - "Vast sums of money ... to keep the controversy alive"--the 1988 BAT memo. PMID- 9396110 TI - Cigar advertising: targeting "baby-boomers" and older adults. PMID- 9396111 TI - Cigarette marketing in Senegal, West Africa. PMID- 9396112 TI - Aussie facts, EZ-letters, blue mould, RJR's home page, and UST's "roar tour". PMID- 9396113 TI - Final report of the Advisory Committee on Tobacco Policy and Public Health. PMID- 9396114 TI - AHCPR smoking cessation guideline: its' goals and impact. PMID- 9396115 TI - AHCPR smoking cessation guideline: a fundamental review. PMID- 9396116 TI - Implementing smoking cessation protocols in medical and dental practices. PMID- 9396117 TI - Implementing the AHCPR guideline in health management organisations. PMID- 9396118 TI - Reaching the medically underserved with the AHCPR guideline. PMID- 9396119 TI - Primary-care physicians. PMID- 9396120 TI - Two pharmacy-practice models for implementing the AHCPR smoking cessation guideline. PMID- 9396121 TI - AHCPR smoking cessation guideline goals and impact: examples from the nursing field. PMID- 9396122 TI - Implications of the AHCPR guideline for psychological practice and research. PMID- 9396123 TI - Smoking cessation initiatives in Minnesota. PMID- 9396124 TI - Employers' and purchasers' roles in smoking cessation. PMID- 9396125 TI - An American Medical Association perspective on smoking cessation guidelines. PMID- 9396126 TI - Consumers and the AHCPR smoking cessation guideline. PMID- 9396127 TI - Changing provider behaviour: provider education and training. PMID- 9396128 TI - Changing physicians' practices. PMID- 9396130 TI - Defining benefits and payment for smoking cessation treatments. PMID- 9396129 TI - Healthcare report cards and tobacco measures. PMID- 9396131 TI - Selling smoking cessation to the states. PMID- 9396132 TI - Pregnant women, infants, and the cost savings of smoking cessation. PMID- 9396133 TI - A managed-care perspective on the AHCPR guideline. PMID- 9396134 TI - Lysosomal storage diseases of animals: an essay in comparative pathology. AB - A wide variety of inherited lysosomal hydrolase deficiencies have been reported in animals and are characterized by accumulation of sphingolipids, glycolipids, oligosaccharides, or mucopolysaccharides within lysosomes. Inhibitors of a lysosomal hydrolase, e.g., swainsonine, may also induce storage disease. Another group of lysosomal storage diseases, the ceroid-lipofuscinoses, involve the accumulation of hydrophobic proteins, but their pathogenesis is unclear. Some of these diseases are of veterinary importance, and those caused by a hydrolase deficiency can be controlled by detection of heterozygotes through the gene dosage phenomenon or by molecular genetic techniques. Other of these diseases are important to biomedical research either as models of the analogous human disease and/or through their ability to help elucidate specific aspects of cell biology. Some of these models have been used to explore possible therapeutic strategies and to define their limitations and expectations. PMID- 9396135 TI - Systemic AA amyloidosis in captive cheetahs (Acinonyx jubatus). AB - Ongoing disease surveillance of necropsied captive cheetahs (Acinonyx jubatus) (n = 141) revealed a high prevalence of renal amyloidosis (n = 54 [38%]; age 1 to 16 years). The prevalence increased from 20% in pre- 1990 necropsies to 70% of cheetahs necropsied in 1995. In 74% of the cheetahs with amyloidosis, renal failure was determined to be the sole or partial cause of death. Papillary necrosis was seen only in affected cheetahs and involved 25% of these animals. Amyloid was present predominantly in the medullary interstitium, with minimal glomerular involvement. The amyloid deposits were immunohistochemically identified as AA type using antisera to both human and canine protein AA. A high percentage (52%) of animals with renal amyloid also had subsinusoidal hepatic AA amyloid deposits. Inflammatory diseases were identified in 100% of affected cheetahs. The most common inflammatory disease was chronic lymphoplasmacytic gastritis. The prevalence and severity of gastritis was higher in cheetahs with amyloidosis, and the prevalence of severe gastritis increased from 16% to 43%, coinciding with the increase in prevalence of amyloidosis. These findings suggest that cheetahs have a high prevalence of systemic amyloidosis in response to inflammation and that renal amyloidosis is an increasingly significant cause of morbidity and mortality in captive cheetah populations. Factors of potential importance in the apparent high prevalence of AA amyloidosis in cheetahs are currently being investigated in our laboratories. PMID- 9396136 TI - Pathobiology of H5N2 Mexican avian influenza virus infections of chickens. AB - To determine the association between specific structural changes in the hemagglutinin gene and pathogenicity of avian influenza viruses (AIVs), groups of 4-week-old White Plymouth Rock chickens were inoculated intravenously or intranasally with AIVs of varying pathogenicities isolated from chickens in central Mexico during 1994-1995. Mildly pathogenic (MP) viruses had a common hemagglutinin-connecting peptide sequence of Pro-Gln-Arg-Glu-Thr-Arg decreases Gly and had restricted capability for replication and production of lesions in tissues. The principle targets for virus replication or lesion production were the lungs, lymphoid organs, and visceral organs containing epithelial cells, such as kidney and pancreas. Death was associated with respiratory and/or renal failure. By contrast, highly pathogenic (HP) AIVs had one substitution and the addition of two basic amino acids in the hemagglutinin connecting peptide, for a sequence of Pro-Gln-Arg-Lys-Arg-Lys-Thr-Arg decreases Gly. The HP AIVs were pantropic in virus replication and lesion production ability. However, the most severe histologic lesions were produced in the brain, heart, adrenal glands, and pancreas, and failure of multiple critical organs was responsible for disease pathogenesis and death. No differences in lesion distribution patterns or in sites of AIV replication were evident to explain the variation in mortality rates for different HP AIVs, but HP AIVs that produced the highest mortality rates had more severe necrosis in heart and pancreas. The ability of individual HP AIVs to produce low or high mortality rates could not be explained by changes in sequence of the hemagglutinin-connecting peptide alone, but probably required the addition of other undetermined genomic changes. PMID- 9396137 TI - Immunohistochemical demonstration of African horse sickness viral antigen in formalin-fixed equine tissues. AB - The distribution of viral antigen was studied in various tissues of three ponies, aged 3-4 years, infected experimentally with a virulent strain of African horse sickness virus (AHSV) serotype 4. Tissues were collected from the animals in the terminal stage of the peracute form of the disease and from one noninfected horse, included as a control. A polyclonal antibody with specificity for AHSV, plus the nonstructural protein NS2, was used in a sensitive avidin-biotin peroxidase-complex (ABC) method performed on formalin-fixed, paraffin-embedded tissue sections. AHSV antigen was located primarily in endothelial cells of capillaries and small venous and arteriolar vessels, particularly of cardiopulmonary tissues. Viral antigen was also identified in cells resembling macrophages and in reticular cells of spleen and lymph nodes. The pattern of viral antigen labeling in some lymph nodes along the mantle zone of lymphoid follicles was compatible with the morphology of cellular processes of follicular dendritic cells. In some tissues, viral antigen was detected occasionally in circulating cells, probably monocytes, within the larger vessels. These findings suggest that endothelial cells, and to a lesser extent mononuclear cells, are the main target cells of AHSV infection during the late stage of the peracute form of the disease. PMID- 9396138 TI - Histopathologic and ultrastructural alterations of white liver disease in sheep experimentally depleted of cobalt. AB - Many cobalt-deficient sheep develop liver lesions known as ovine "white liver" disease, but the etiology of these changes is controversial. It has been suggested that cofactors are required for development of liver damage in cobalt deficient sheep. In this study, one group of lambs (n = 5) was fed a diet low in cobalt (4.5 micrograms/kg) while a group of control lambs (n = 4) received the same diet after it had been supplemented with cobalt (1000 micrograms/kg). All cobalt-depleted lambs had reduced growth rate, anorexia, lacrimation, and alopecia, and they eventually became emaciated (mean body weight at end of study: 83% of initial body weight). Plasma concentrations of bilirubin and serum activity of glutamate-oxaloacetate transferase were elevated in these animals, while plasma concentrations of vitamin B12 were reduced (less than 220 pmol/L from day 42). Fatty degeneration of the liver associated with reduced concentrations of vitamin B12 (14.5 pmol/g) was seen in these animals at necropsy at 196 days. Microscopic liver lesions included accumulation of lipid droplets and lipofuscin particles in hepatocytes, dissociation and necrosis of hepatocytes, and sparse infiltration by neutrophils, macrophages, and lymphocytes. Ultrastructural hepatocytic alterations included swelling, condensation and proliferation of mitochondria, hypertrophy of smooth endoplasmic reticulum, vesiculation and loss of arrays of rough endoplasmic reticulum, and accumulation of lipid droplets and lipofuscin granules in cytoplasm of hepatocytes. No liver lesions were seen in control lambs. The results of this study indicate that cofactors are not a prerequisite to development of hepatic damage in cobalt-deficient sheep. Reduced activities of the vitamin B12-dependent enzymes, methylmalonyl CoA mutase and methionine synthase, and lipid peroxidation are of likely pathogenetic importance in the development of the lesions. PMID- 9396139 TI - In situ hybridization demonstration of albumin mRNA in B6C3F1 murine liver and hepatocellular neoplasms. AB - In situ hybridization was used to detect albumin mRNA in normal liver and hepatocellular neoplasms in 20 male B6C3F1 mice between 17 and 24 months of age. Positive signals for albumin were observed consistently in the cytoplasm of hepatocytes in normal liver, particularly in periportal areas. No signals were observed in other cells, such as Kupffer's cells, mesenchymal cells, or bile duct epithelium. Of hepatocellular adenomas, 11/11 (100%) stained positively for albumin mRNA, whereas 14/15 (93%) of primary hepatocellular carcinomas showed positive expression. Albumin mRNA was also detected in extrahepatic metastases of hepatocellular carcinoma, including 9/15 (60%) of pulmonary neoplasms and 5/12 (42%) of metastases at other sites. The pulmonary metastases of hepatocellular carcinoma frequently exhibited a glandular, papillary, or sarcomatous histologic appearance. The presence of albumin in these tumors, lacking characteristic hepatocellular phenotype, is a potential determinant of hepatic lineage. We conclude that in situ hybridization for albumin mRNA in mice is a useful tool in the differential diagnosis of hepatocellular carcinoma, particularly in the case of pulmonary metastasis. This technique may also enable recognition of hepatocyte differentiation in glandular structures with phenotypic features of biliary cells, as seen in mixed hepatocellular-cholangial neoplasms. PMID- 9396140 TI - Regression of pulmonary lesions produced by inhaled titanium dioxide in rats. AB - Inhaled ultrafine particles of TiO2 (TiO2-D, 20 nm particle size) lead to a greater pulmonary inflammatory response than larger pigment-grade particles (TiO2 F, 250 nm). Male Fisher 344 rats were exposed for 6 hours a day, 5 days a week, for 3 months to 1) filtered air (control); 2) TiO2-F, 22.3 mg/m3; 3) TiO2-D, 23.5 mg/m3; or 4) crystalline SiO2, a positive control particle (approximately 800 nm particle size, 1.3 mg/m3). Groups of 3-4 animals were sacrificed at 6 and 12 months following the completion of exposure. Pulmonary effects of exposure were evaluated using standard hematoxylin and eosin-stain sections, histochemical stains for collagen, and immunohistochemical assays for cell turnover. Six months after animals were exposed to SiO2, they had moderate focal interstitial fibrosis and moderately severe focal alveolitis. Animals exposed to TiO2-D had slightly less fibrosis. The least fibrosis was seen in the TiO2-F group. At 1 year after exposure, fibrosis was still present but decreased in the SiO2 group. The amount of interstitial fibrosis in the TiO2-D- and TiO2-F-treated animals had largely returned to untreated control levels, although an increased number of alveolar macrophages persisted, usually with retained particles. There was discordance between bromodeoxyuridine and proliferating cell nuclear antigen indices, most probably due to cytokine elaboration in the areas of inflammation, which may have altered the expression of proliferating cell nuclear antigens. There was no detectable fibroblast labeling at the 6-month observation and only very low levels at 12 months. Thus, although initially irritant, TiO2-induced lesions regressed during a 1-year period following cessation of exposure. PMID- 9396141 TI - Intercellular adhesion molecule-1 expression in dextran sodium sulfate-induced colitis in rats. AB - Orally administered dextran sodium sulfate (DSS) produces an acute colitis in rodents. The pathogenesis is unknown but may relate to DSS-mediated toxicity of colonic crypt epithelium and/or DSS-induced inflammation. The purpose of this study was to determine when colonic mucosal inflammation, as indicated by histopathology and intercellular adhesion molecule-1 (ICAM-1) expression, occurs relative to crypt epithelial damage. Groups of eight adult male Wistar rats were administered 5.0% DSS solution in the drinking water for 2-6 days. Clinical signs at 3 days consisted of loose stool, progressing to marked rectal hemorrhage by days 5 and 6 that correlated with marked intraluminal colonic hemorrhage at necropsy. Histological lesions of predominantly the distal colon consisted of multifocal areas of mucosal erosion, reduction in goblet cells, dilated crypts, crypt collapse, increased lamina propria neutrophils, and submucosal edema on days 2 and 3, progressing to locally extensive ulceration and marked mixed inflammatory infiltrates by days 4-6. Enhanced expression of ICAM-1, demonstrated by both immunohistochemical and northern blot analysis, was evident in colonic mucosa as early as day 2, with consistent increases through days 3-6. Results demonstrate that enhanced colonic mucosal endothelial cell ICAM-1 expression is an early event in the inflammatory cascade of DSS-induced colitis. PMID- 9396142 TI - Lesions of aryl-hydrocarbon receptor-deficient mice. AB - We have analyzed the possible role of the aryl-hydrocarbon receptor (AHR) in the aging process of mice using a homozygous null mouse (Ahr-/-) line as a model. We studied 52 male and female Ahr-/- mice aged from 6-13 months. Forty-six percent died or were ill by 13 months of age. Ahr-/- mice developed age-related lesions in several organs, some of which were apparent after only 9 months of age. Cardiovascular alterations included cardiomyopathy (100%) with hypertrophy and focal fibrosis. Vascular hypertrophy and mild fibrosis were found in the portal areas of the liver (81%), and vascular hypertrophy and mineralization were common in the uterus (70%). Gastric hyperplasia that progressed with age into polyps was evident in the pylorus of 71% of the mice over 9 months of age. Ahr-/- mice had T cell deficiency in their spleens but not in other lymphoid organs. The immune system deficiency described previously could be the origin for the rectal prolapse found in 48% of the null mice, associated with Helicobacter hepaticus infection. In the dorsal skin (53% incidence), severe, localized, interfollicular and follicular epidermal hyperplasia, with hyperkeratosis and acanthosis, and marked dermal fibrosis, associated with the presence of anagenic hair follicles, were also evident. None of these lesions were found in 42 control (Ahr +/+ or +/ ) mice of similar ages. These observations suggest that the AHR protein, in the absence of an apparent exogenous (xenobiotic) ligand, plays an important role in physiology and homeostasis in major organs in mice, and further supports an evolutionary conserved role for this transcription factor. PMID- 9396143 TI - S-100 immunoreactivity in melanomas of two marsupials, a bird, and a reptile. AB - S-100 proteins are abundant in melanocytes of the skin; thus, S-100 immunoreactivity has been used as a diagnostic criterion for melanoma in humans and other placental mammals. We tested cutaneous melanomas of two marsupials, a bird, and a snake for S-100 immunoreactivity, using a polyclonal rabbit antibovine S-100 antibody. The tumor from a Tasmanian Pademelon (Thylogale billaridierii) was composed of large epithelioid cells, most of which had S-100 positive cytoplasm. In general, there were only scattered individual spindle shaped S-100-positive cells or groups of cells in the primary mass from a Spotted tailed Quoll (Dasyurus maculatus); S-100 staining was primarily nuclear. Cells comprising the melanomas of the Australian Cormorant (Phalacrocorax carbo) and the Death Adder (Acanthophis antarcticus) were S-100-negative, although peripheral nerve bundles in both were S-100-positive. PMID- 9396144 TI - Botryoid-type embryonal rhabdomyosarcoma of liver in a young cat. AB - An unusual malignant mesenchymal tumor arising in the liver of a 2-year-old cat is described. Histologically, the tumor showed considerable variation in growth pattern, cellularity, and cell types. Phenotypical diversity was confirmed by immunohistochemistry, showing expression of desmin, vimentin, S-100, and neuron specific enolase in various areas of the tumor. On the basis of histopathology, immunohistochemistry, electron microscopy, and gross morphology, the tumor was classified as botryoid-type embryonal rhabdomyosarcoma. Differential diagnosis included so-called undifferentiated (embryonal) sarcoma of the liver, a rare tumor of the pediatric age group in humans. Problems of tumor heterogeneity and differentiation in mesenchymal tumors are discussed. PMID- 9396145 TI - Primary encephalic plasma cell tumor in a dog. AB - A 5-year-old female spayed Spitz dog had a 5-week history of right head tilt, seizures, and progressive quadriplegia. Analysis of cerebrospinal fluid revealed 27,600 white blood cells per microliter with 63% mononuclear phagocytes, 27% lymphocytes, 6% neutrophils, 3% plasmacytoid cells, and 1% eosinophils, and over 2000 mg/dl protein. On contrast-enhanced magnetic resonance images, a focal 1-cm oval lesion was identified in the right ventral brainstem. There was also marked contrast enhancement of the meninges in the following areas: surrounding the brainstem, outlining cerebellar folia, along the ventral floor of the brain and extending to the falx cerebri, and partially outlining the left frontal lobe. At necropsy, the areas of contrast enhancement corresponded to the presence of compact cellular sheets of pleomorphic, anisocytotic, oval to polygonal neoplastic cells with plasmacytoid differentiation. The smaller of these plasmacytoid cells stained predominantly for cytoplasmic immunoglobulin A using immunoperoxidase methodology. Ultrastructurally, the neoplastic cells had morphologic features typical of plasma cells, with large amounts of predominantly rough endoplasmic reticulum with variably prominent Golgi formation. This is the first report of a canine primary intracranial malignant plasma cell tumor. PMID- 9396146 TI - Two cyclohexanespiro-5'-hydantoin monohydrates. AB - Cyclohexanespiro-5'-hydantoin monohydrate, C8H12N2O2.H2O, has a chair-shaped cyclohexane ring with endocyclic torsion-angle magnitudes in the range 54.4 (2) 56.3 (2) degrees. All potential hydrogen-bond donors are involved in intermolecular hydrogen bonding, with lengths in the range 2.760 (2)-2.908 (2) A. In its indolyl adduct, 2-(3-indolyl)cyclohexanespiro-5'-hydantoin monohydrate, C16H17N3O2.H2O, the cyclohexane moiety adopts a chair conformation with the indolyl substituent in an equatorial position. The N-H portion of the hydantoin ring is cis to indolyl, while the C=O of the hydantoin is trans. Endocyclic torsion-angle magnitudes of the cyclohexane ring are in the range 54.2 (2)-56.7 (2) degrees. All potential hydrogen-bond donors are involved in intermolecular hydrogen bonds, with lengths 2.828 (2)-3.187 (2) A. PMID- 9396147 TI - Colocalization of microtubules and mitochondria in the yeast Schizosaccharomyces japonicus var. versatilis. AB - Both living and fixed cells of Schizosaccharomyces japonicus var. versatilis showed thread-like mitochondria when studied by phase-contrast and fluorescence microscopy. In the interphase cells, mitochondria extended from pole to pole and converged towards the growing tips. The mitochondrial threads did not disrupt but persisted during mitosis and, subsequently, their bundle was split between the two daughter cells by a concentrically growing septum. Mitochondria in the interphase cells were accompanied by cytoplasmic microtubules. These disappeared during mitosis and, instead, spindle microtubules were formed in the nucleus. The cytoplasmic microtubules reappeared after anaphase B, again in coaligment with mitochondria. Protoplasting as well as the action of microtubule inhibitors methyl-1-(butylcarbamoyl)-2-benzimidazolecarbamate (benomyl) and 2 methylbenzimidazole (MBC) resulted in rapid disintegration of microtubules and, suprisingly, also in disruption of mitochondria into small bodies. Removal of the inhibitors or a short regeneration of protoplasts allowed both the cytoplasmic microtubules and the thread-like mitochondria to reaggregate into the original pattern. Cytochalasin D treatment caused a complete disintegration of actin filaments, while the cytoplasmic microtubules and mitochondria remained intact. These findings of a transient close association of mitochondria and microtubules and their relative independence of the arrangement of actin filaments suggest that microtubules, but not actin cables, form supports for positioning or movement of mitochondria along the cylindrical cells. The persistence of mitochondria in the cell centre during mitosis may be accounted for by the fact that disrupted microtubules fail to provide support for mitochondrial movement towards the cell poles. PMID- 9396148 TI - Influence of altered plasma membrane fatty acid composition on cesium transport characteristics and toxicity in Saccharomyces cerevisiae. AB - The influence of altered plasma membrane fatty acid composition on cesium uptake and toxicity was investigated in Saccharomyces cerevisiae. Detailed kinetic studies revealed that both the Vmax and Km values for Cs+ transport increased (by approximately twofold in the latter case) when S. cerevisiae was grown in medium supplemented with the polyunsaturated fatty acid linoleate. In addition, Cs+ uptake by linoleate-enriched cells was considerably less sensitive to the competitive effects of other monovalent cations (K+, Rb+, and NH4+) than that by unsupplemented cells. Stimulation of Cs+ uptake in the presence of certain K+ and Rb+ concentrations was only evident in linoleate-enriched S. cerevisiae. At 100 mM CsCl, the initial rate of Cs+ uptake was greater in linoleate-supplemented cells than in unsupplemented cells and this was reflected in a more rapid displacement of cellular K+. However, little difference in net Cs+ accumulation between linoleate-supplemented and unsupplemented cells was evident during prolonged incubation in buffer or during growth. Thus, Cs+ toxicity was similar in linoleate-supplemented and unsupplemented cells. The results were consistent with the Cs+ (K+) transport mechanism adopting an altered conformational state in linoleate-enriched S. cerevisiae. PMID- 9396149 TI - Tyrosine is involved in protection from oxidative stress in Saccharomyces cerevisiae. AB - The phenotypic characterization of a Saccharomyces cerevisiae mutant unable to grow under agitated conditions is presented here. When this strain was incubated under aerobic conditions, it did not grow and the viability of the culture decreased. The loss in viability was prevented by the addition of antioxidants or chelating agents to the medium, indicating that this mutant was unable to withstand the oxidative stress generated by aerobic metabolism. This strain was complemented with plasmids from a yeast genomic library. The transformants that were obtained carried plasmids harbouring the TYR1 gene, which codes for one of the enzymes involved in tyrosine biosynthesis. A monogenic S. cerevisiae tyr1 mutant obtained from the Yeast Genetic Stock Center showed higher sensitivity to hydrogen peroxide than a TYR1 strain. This sensitivity was reverted when this strain was complemented with the TYR1 gene. Considering these results, we propose that tyrosine plays a role in the protection against oxidative stress. PMID- 9396150 TI - Functional analysis of sigma-70 consensus promoters in Pseudomonas aeruginosa and Escherichia coli. AB - A series of synthetic promoters, based upon the Escherichia coli sigma 70 consensus promoter sequence, was constructed upstream of the lacZ reporter gene in the modified broad-host-range vector pQF52. The role of the intervening spacer region in gene expression in Pseudomonas aeruginosa and E. coli was studied by insertions and deletions within this region. In P. aeruginosa and E. coli the patterns of gene expression were identical with maximum beta-galactosidase activity being measured from promoters possessing 19 bp in their intervening regions, presumably as a result of impeded promoter clearance with the consensus 17-bp promoter. In P. aeruginosa a second occurrence of enhanced activity, which could not be attributed to the involvement of the alternative sigma factor RpoN (sigma 54), was evident with the promoter having a 16-bp spacer. PMID- 9396151 TI - Change of mechanical activity to contraction from the relaxation induced by the intracellular Ca2+ antagonist KT-362; effects of alkylation of side chain, and substitution of 2,3,4,5-tetrahydro-1,5-benzothiazepine derivatives. AB - KT-362 (5-[3-[2-(3,4-Dimethoxyphenyl)ethyl]aminopropionyl]-2,3,4, 5-tetrahydro 1,5-benzothiazepine fumarate) is an intracellular Ca2+ antagonist. The compound obtained by introducing methyl groups onto the nitrogen (R2) of the side chain of KT-362 showed vasoconstrictive activity. Therefore we synthesized various derivatives, and examined their activities. Substitution at position R2 of the side chain resulted in potent contractile activity, and the optimal alkyl length was two or three carbons. The potency was further increased by the introduction of a chloro group at the R1 position of 2,3,4,5-tetrahydro-1,5-benzothiazepines. One of the synthesized compounds, 8-chloro-5-?N-ethyl-N-[2-(3,4 dimethoxyphenyl)ethyl]aminopropionyl?-2,3,4, 5-tetrahydro-1,5-benzothiazepine fumarate (9b), showed an EC50 value of 3.47 x 10(-8) M for contraction of rabbit iliac artery. The action of compound 9b was antagonized competitively by an H1 histamine receptor antagonist, diphenhydramine, and the pA2 value was 7.82. The maximum constriction was inhibited by a Ca2+ entry blocker, nicardipine, but not by an alpha 1-adrenoreceptor antagonist, prazosin. In a Ca(2+)-free medium, tonic constriction induced by 9b disappeared, and only a phasic constriction was observed. Though this phasic constriction was inhibited by diphenhydramine, it was not inhibited by prazosin or nicardipine. PMID- 9396153 TI - Synthesis and antiplatelet evaluation of alpha-methylene-gamma-butyrolactone bearing 2-methylquinoline and 8-hydroxyquinoline moieties. AB - In a search for inhibitors of platelet aggregation, some alpha-methylene-gamma butyrolactones bearing 2-methylquinoline and 8-hydroxyquinoline moieties were synthesized and evaluated for antiplatelet activities against thrombin (Thr)-, arachidonic acid (AA)-, collagen (Col)-, and platelet-activating factor (PAF) induced aggregation in washed rabbit platelets. With the exception of 2-[[2,3,4,5 tetrahydro-4-methylene-5-oxo-2-(4-phenylphenyl)-2 -furanyl]methoxy]-8 hydroxyquinoline (8f), these alpha-methylene-gamma-butyrolactones completely inhibited the platelet aggregation induced by AA and Col. The 2-methylquinoline derivatives were also active against Thr- and PAF-induced aggregation, while their 8-hydroxyquinoline counterparts were relatively inactive. PMID- 9396152 TI - Pyrrole butyric acid derivatives as inhibitors of steroid 5 alpha-reductase. AB - A series of pyrrole butyric acid derivatives was synthesized and evaluated for inhibitory activity on human and rat steroid 5 alpha-reductase in vitro and ex vivo. 3-Benzoyl-4-alkylpyrrole-1-butyric acids and 1-methyl-2-alkyl-3 benzoylpyrrole-5-butyric acid derivatives were effective inhibitors. Structure activity relationships were evaluated among the 37 compounds synthesized. Compound 37 (HQL-1069) shows potent inhibitory activities against both rat and human 5 alpha-reductase. PMID- 9396154 TI - Synthesis and antinociceptive activity of [D-Ala2]Leu-enkephalin derivatives conjugated with the adamantane moiety. AB - Based on the physicochemical and pharmacological properties of drugs having an adamantane skeleton, an adamantane-based moiety was evaluated as a drug carrier for poorly absorbed compounds, including peptides, active towards the central nervous system (CNS). Seven [D-Ala2]Leu-enkephalin derivatives conjugated with an adamantane-based moiety at the C-terminus or N-terminus were prepared by the solution-phase method and their biological activities were examined. The compounds derivatized at the C-terminus through an ester or amide linkage were much more lipophilic than the parent peptide and exhibited moderate in vitro opioid activity (guinea-pig ileum assay). Among them, four derivatives (1, 2, 4, 5), exhibited significant antinociceptive effects in an in vivo assay (mouse tail pressure test) after subcutaneous administration. This result suggests that the introduction of the lipophilic adamantane moiety into [D-Ala2]Leu-enkephalin would improve the permeation of the poorly absorbed parent peptide through the blood-brain-barrier (BBB) without loss of antinociceptive effect. PMID- 9396155 TI - Amino acids and peptides. XXX. Preparation of Arg-Gly-Asp (RGD) hybrids with poly(ethylene glycol) analogs and their antimetastatic effect. AB - Hybrids of a fibronectin-related peptide[Arg-Gly-Asp (RGD)] with poly(ethylene glycol) (PEG) analogs were prepared by a simple and easy procedure. Two amino-PEG analogs were used as carriers for hybrid formation of the RGD. One was poly(oxyethylene)dipropylamine and the other was Jeffamine ED type, which has branched chains. RGD peptides were formed stepwise on PEG analogs by the diisopropylcarbodiimide method. The synthetic intermediates were easily purified by molecular-sieve gel chromatography and the final products were purified by molecular-sieve gel chromatography, followed by HPLC. This simple and easy preparation procedure using molecular-sieve gel chromatography for purification of synthetic intermediates is advantageous for the preparation of peptide-polymer hybrids. We found that PEG is stable to HF treatment at 0 degree C for 1 h. The inhibitory effect of the RGD hybrids on experimental metastasis of B16-BL6 was examined in mice. The Jeffamine type hybrid showed no inhibitory effect at the dose of 1 mg/mouse, but poly(oxyethylene)dipropylamine type hybrid was inhibitory at the same dose. The effect of the latter hybrid was about the same as that of 1 mg of RGD. One mg of the hybrid contains 0.18 mumol of RGD and 1 mg of RGD is 2.38 mumol. Thus it can be said that the inhibitory effect of RGD was potentiated by hybrid formation with poly(oxyethylene)diisopropylamine. PMID- 9396156 TI - Synthesis and structure-activity relationships of novel 2',2'-difluoro analogues of docetaxel. AB - To investigate the role of the 2'-hydroxy group at the C-13 side chain of docetaxel in the antitumor activity, we prepared several 2',2'-difluoro derivatives of docetaxel and evaluated their cytotoxicity against mouse leukemia and human tumor cell lines and their microtubule disassembly-inhibitory activity. These analogues were prepared by esterification of protected 10-deacetylbaccatin III (21) with appropriate alpha, alpha-difluorinated carboxylic acids (Charts 1 and 2). Among these 2',2'-difluorodocetaxel derivatives, 2',2'-difluorodocetaxel (23b) was approximately 3-10 times as active as 2'-fluorodocetaxel (29a) in terms of cytotoxicity. In addition, the 3'-(2-furyl) (23h) and 3'-(2-pyrrolyl) (23p) analogues showed activity comparable or superior to that of docetaxel (2). PMID- 9396157 TI - Polyenylidene thiazolidinedione derivatives with retinoidal activities. AB - Several polyenylidene thiazolidine or 2-thioxo-4-thiazolidinone derivatives were synthesized and their retinoidal activities were examined in terms of the differentiation-inducing ability towards human promyelocytic leukemia HL-60 cells and inhibitory effect on interleukin (IL)-1 alpha-induced IL-6 production in MC3T3-E1 cells. Compounds containing a trimethylcyclohexenyl ring induced HL-60 cell differentiation with weaker activity than retinoic acid (1a) by one or two orders of magnitude. The thiazolidinedione derivatives (2, 5, 7) showed stronger activity than the corresponding 2-thioxo-4-thiazolidinone derivatives (3, 6, 8). The effects of a retinoid antagonist (LE540) and synergists (retinoid X receptor (RXR) agonists, HX600 or HX630) on the activities of thiazolidine derivatives indicate that these compounds elicit their activities through the nuclear retinoic acid receptors (RARs). All the thiazolidines examined also inhibited IL 1 alpha-induced IL-6 production with IC50 values of 10 nM order. The retinoidal activities of the thiazolidines are significant, considering that replacement of the carboxylic acid in retinoid structures with bioisosteric functional groups is generally ineffective, as seen in the structure-activity relationships of retinoidal benzoic acids. PMID- 9396158 TI - Fluorometric determination of 1,2,3,4-tetrahydro-6,7-dihydroxyisoquinoline in biological materials by HPLC. AB - In the belief that endogenous 1,2,3,4-tetrahydro-6,7-dihydroxyisoquinoline (DA Fp) could be a potential marker involved in the etiology of various diseases such as Parkinsonism, we attempted to develop a fluorescence method for DA-Fp. It was synthesized by condensation of dopamine with formaldehyde according to an established method. Periodate was identified by screening from various oxidation reagents as a fluorescence reagent to DA-Fp. Optimal reaction conditions were obtained with 0.25 mM NaIO4 in 0.1 M phosphate buffer (pH 8.0) at 37 degrees C for 15 min. The fluorescence spectrum of the derivative showed that we had found a new reaction specific for DA-Fp. This reaction was coupled on-line to high performance liquid chromatography (HPLC), which enabled us to achieve a highly sensitive method for determining DA-Fp. A working curve was linear from 2 to 800 pmol of DA-Fp per injection. To determine DA-Fp in biological materials, the pretreatment before HPLC was optimized by hydrolysis of its conjugate and suppression of the artifact with l-phenylephrine. Urinary excretion of DA-Fp in men was measured by this new present method. The urinary excretion of endogenous DA-Fp increased in a rabbit given L-DOPA. The DA-Fp concentration was determined in rat brain. The significance of DA-Fp in these biological materials is discussed and evaluated. We conclude that the present method will be useful for studying tetrahydroisoquinolines involved in many diseases. PMID- 9396159 TI - Nitroarene concentrations and direct-acting mutagenicity of diesel exhaust particulates fractionated by silica-gel column chromatography. AB - Diesel exhaust particulates were extracted with benzene-ethanol (3:1, v/v) and separated into five fractions by silica-gel column chromatography. Direct-acting mutagenic activity was assayed by the Ames test using the Salmonella typhimurium YG1024 strain. The total activity of five fractions was about four times greater than that of the crude extract, suggesting that the activities in the fractions were suppressed in the crude extract. Strong activity was observed in fraction 4 which was eluted with dichloromethane (61.5% of the total activity) and fraction 5 which was eluted with ethanol (35.3%). Nitropolycyclic aromatic hydrocarbons (NPAHs) were determined by high-performance liquid chromatography with chemiluminescence detection. They were found mainly in fraction 4, although one NPAH was in fraction 3 which was eluted with n-hexane-dichloromethane (3:1, v/v). Based on these results, 53.1% of the activity in fraction 4 was attributed to NPAHs. The contribution of 1-nitropyrene and 1,3-, 1,6- and 1,8-dinitropyrenes was great and that of the other NPAHs was small. The mutagenic compound in fraction 5 was not identified. Fractions 1 and 2, which were eluted with n hexane, and fraction 3 suppressed the activity of fraction 4. Polycyclic aromatic hydrocarbons in fractions 2 and 3 were considered as possible suppressors of NPAHs. PMID- 9396160 TI - Hepatoprotective principles of Swertia japonica Makino on D galactosamine/lipopolysaccharide-induced liver injury in mice. AB - The n-BuOH extract of Swertia japonica showed a significant hepatoprotective effect on D-galactosamine (D-GalN)/lipopolysaccharide (LPS)-induced liver injury in mice. The activity-guided fractionation led to the isolation of a new tetrahydroxanthone derivative, tetrahydroswertianolin (1), as well as two known iridoids, gentiopicroside (2) and sweroside (3). Their structures were elucidated by spectroscopic methods and chemical reactions. Of the three compounds, 2 and 3 possessed mild hepatoprotective activity at a dose range of 25-50 mg/kg, whereas, 1 exhibited potent activity in a dose-dependent manner. The hepatoprotective effect of tetrahydroswertianolin (1) was stronger than that of glycyrrhizin which was used as a positive control. PMID- 9396162 TI - Degradation of a novel tripeptide, tert-butoxycarbonyl-Tyr-Leu-Val-CH2Cl, with inhibitory effect on human leukocyte elastase in aqueous solution and in biological fluids. AB - The stability of tert-butoxycarbonyl-Tyr-Leu-Val-CH2Cl (YLV) with inhibitory effect on human leukocyte elastase was investigated in aqueous solution, alpha chymotrypsin solution and biological media. In all cases studied here, the degradation was observed as a pseudo-first order reaction. The half-life for the degradation of YLV in an aqueous solution of pH 7.4 at 37 degrees C was 35.9 h. YLV was most stable at about pH 3.8-5.8 and the effect of temperature was explained by the Arrhenius equation. The activation energies of the degradation in aqueous solutions at pH 2.0, 4.8, and 7.4 were 24.6, 22.1 and 23.4 kcal/mol, respectively. The degradation products in aqueous solution were analyzed by HPLC MS and were estimated as Boc-Tyr-Leu-Val-CH2OH at pH 7.4 and H2N-Tyr-Leu-Val CH2Cl at pH 2.0. In a bovine pancreas alpha-chymotrypsin solution at 37 degrees C, the half-life of YLV was 15 min at 25.6 micrograms/ml of alpha-chymotrypsin solution. In the rat plasma, the half-life of YLV was 42.4 min (YLV 26.7 micrograms/ml plasma), and in rat liver, lung and spleen homogenates, the degradation rate constants of YLV were 37.6, 10.3 and 23.5 times larger than that in plasma solution, respectively (all fluids containing 5 mg protein/ml). YLV was less stable than nafarelin acetate, secretin, adrenocorticotropic hormone (ACTH) and gonadorelin in an aqueous solution of pH 7.4. PMID- 9396163 TI - Nonpeptide arginine vasopressin antagonists for both V1A and V2 receptors: synthesis and pharmacological properties of 2-phenyl-4'-[(2,3,4,5-tetrahydro-1H-1 benzazepin-1-yl)carbonyl]benzanil ide derivatives. AB - A series of compounds structurally related to 4'-[(2,3,4,5-tetrahydro-1H-1 benzazepin-1-yl)carbonyl]benzanili de was synthesized and demonstrated to have arginine vasopressin (AVP) antagonist activity for both V1A and V2 receptors. The introduction of a phenyl or a 4-substituted phenyl group into the ortho position of the benzoyl moiety resulted in an increase in both binding affinity and antagonistic activity. The 2-(4-methylphenyl) derivative (1g) exhibited high antagonistic activities for both V1A (8.6-fold) and V2 (38-fold) receptors and high oral activity (8.6-fold) compared with the 2-methyl lead compound (1a). Detail of the synthesis and the pharmacological properties of this series are presented. PMID- 9396164 TI - Internal structure of phonetic categories: effects of speaking rate. AB - Many studies have shown that listeners process speech in a rate-dependent manner, altering the location of phonetic category boundaries in accord with the acoustic consequences of a change in rate during speech production. In a recent series of papers that focused on a voicing contrast, we reported that the perceptual adjustment for rate is not limited to the region of the category boundary, but extends to well within the category, producing a change in which stimuli are perceived to be the best category exemplars. In the current paper, we provide evidence for the generality of this effect by showing analogous results for a /b/ /w/ contrast, specified by transition duration. The implications of these findings for models of rate-dependent processing are discussed. PMID- 9396165 TI - Exploring the relationship of inspiration duration to utterance duration. AB - Previous work has indicated that there may be a positive relationship between the duration and extent of inspiration and the length of an upcoming utterance. However, none of that work has uniquely implied a role of planning. We attempted to avoid some of the alternative explanations by forcing subjects to utter single sentences ranging in length from 5 to 82 syllables (mean of 27), after inspiring fully and then expiring down to a set level before uttering the sentence. For all 3 subjects, there was a significant positive relationship between utterance length and inspiration duration, regardless of whether inspiration was measured physiologically or acoustically. The 2 subjects with the higher correlations in the articulatory measures also expended air more quickly during the shorter sentences than longer ones, while the other subject had no correlation with exhalation rate. Complexity of the sentence, calculated as the number of clauses in the sentence, did not affect inspiration duration. The individual differences need further investigation, but there is a positive correlation between the duration of the sentence to be said and the inspiration before it when the speaker is required to read sentences while using only one breath. PMID- 9396166 TI - Silent mandibular oscillations in vocal babbling. AB - Early babbling has been characterized as being fundamentally a mandibular oscillation: the infant's repeated lowering and raising of its mandible yields a perceived contrast between consonants produced in a closed vocal tract configuration and vowels produced with an open tract. We wondered whether babblers produce rhythmic mandibular oscillations without phonation and, if so, whether there might be a relationship between such 'jaw wags' and early speech. We report two studies: the first is a longitudinal, observational study of 14 infants, some of whom were hearing and some Deaf. Seven infants (3 hearing, 3 Deaf, and 1 hearing-impaired) produced numerous speech-like, rhythmic jaw wags without phonation; sometimes jaw wags formed a single utterance with phonated babbling. Most jaw wags reported here were produced when these infants were ages 8-13 months. The second study, a survey of 90 parents of 4- to 10-month-old hearing infants, suggests that silent babbles may be a widespread phenomenon of early speech development. PMID- 9396167 TI - Production constraints on utterance-final consonant characteristics in babbling. AB - In order to evaluate hypotheses regarding production constraints on final consonants in babbling, 721 utterance-final consonants produced by 6 infants in consonant-vowel-consonant (CVC) syllables were examined and compared with the preceding consonant in the CVC. Consistent with earlier studies, major patterns were observed for each of the three main consonantal properties--place and manner of articulation and voicing. These patterns included a strong tendency for final consonants to repeat the place of articulation of nonfinal consonants and a tendency for relatively more fricative, nasal and voiceless consonants to occur in final position than in nonfinal position. The high frequency with which final consonants shared place of articulation with the preceding consonant was considered to reflect 'frame dominance' or the tendency of a relatively constant mandibular cycle (the frame) to determine the structure of utterances with very little contribution from other active articulators. The manner and voicing effects were attributed to an overall terminal energy decrease in the vocal production system. PMID- 9396168 TI - Active compost biofiltration of toluene. AB - Composting of leaves and alfalfa (i.e. active compost) was used for the biofiltration of toluene-contaminated air in a 6-L biofilter (initial bed height: 180 mm). During the thermophilic phase (45 to 55 degrees C), toluene biodegradation rates reached 110 g toluene.m-3.h-1 at an inlet concentration of about 5 g.m-3 and a gas residence time of 90 seconds. The highest rates were obtained in the thermophilic phase suggesting a microbial adaptation was occurring. Biodegradation rates decreased rapidly (50% in 48 h) in the cooling stage. Under mesophilic conditions, the maximum biodegradation rates that could be obtained by increasing the inlet toluene concentration were near 89 g toluene.m-3.h-1 which is similar to that reported in the literature for mature compost biofilters. No volatile by-product was detected by gas chromatherapy. Mineralization of 14C-toluene and benzene showed that they were completely degraded into CO2 and H2O under both thermophilic and mesophilic conditions. Bacteria isolated from late mesophilic stage had the capacity to degrade all BTEX compounds but were not able to transform chlorinated compounds. No organisms were isolated which could use toluene as their sole source of carbon and energy at 50 degrees C. Active compost biofiltration should be an excellent process for the treatment of gaseous BTEX by biofiltration. This is the first report of thermophilic biofiltration of toluene. PMID- 9396169 TI - Water stress effects on toluene biodegradation by Pseudomonas putida. AB - We quantified the effects of matric and solute water potential on toluene biodegradation by Pseudomonas putida mt-2, a bacterial strain originally isolated from soil. Across the matric potential range of 0 to -1.5 MPa, growth rates were maximal for P. putida at -0.25 MPa and further reductions in the matric potential resulted in concomitant reductions in growth rates. Growth rates were constant over the solute potential range 0 to -1.0 MPa and lower at -1.5 MPa. First order toluene depletion rate coefficients were highest at 0.0 MPa as compared to other matric water potentials down to -1.5 MPa. Solute potentials down to -1.5 MPa did not affect first order toluene depletion rate coefficients. Total yield (protein) and carbon utilization efficiency were not affected by water potential, indicating that water potentials common to temperate soils were not sufficiently stressful to change cellular energy requirements. We conclude that for P. putida: (1) slightly negative matric potentials facilitate faster growth rates on toluene but more negative water potentials result in slower growth, (2) toluene utilization rate per cell mass is highest without matric water stress and is unaffected by solute potential, (3) growth efficiency did not differ across the range of matric water potentials 0.0 to -1.5 MPa. PMID- 9396170 TI - Secoiridoids from Gentiana siphonantha. AB - Repeated fractionations of the methanol extract of the subterranean parts (rhizomes and roots) of Gentiana siphonantha afforded two new and five known secoiridoids, in addition to the widespread plant constituents beta-sitiosterol, daucosterol and oleanolic acid. The structures of the new acyl secoiridoid glycosides were elucidated as 6'-gentisoyl 8-epikingiside and 2'-gentisoyl gelidoside mainly by a combination of high field NMR techniques. The known secoiridoids were identified as gentiolactone, gentiopicroside, sweroside, gelidoside and trifloroside. None of these constituents was active against human pathogenic fungi (Candida albican, Aspergillus flavus and Trichoderma viride). The chemotaxonomic significance of the isolates is discussed briefly. PMID- 9396171 TI - Marasmane sesquiterpenes from the basidiomycete Clitocybe hydrogramma. AB - Investigations of solid cultures of Clitocybe hydrogramma gave rise to the isolation of two new alpha, beta-unsaturated dialdehydes having the marasmane skeleton. Their structures have been determined on the basis of 1H and 13C NMR evidence. PMID- 9396172 TI - Helicobacter pylori and gastric cancer. An endoscopic series. AB - In a series of 92 patients with gastric cancer who had biopsies of the antrum and fundus, we compared the 65 patients with Helicobacter pylori (71%) with the 27 negative patients. No difference was observed for age, gender or histology (intestinal or diffuse). Significant differences concerned location (11% of H. pylori positive patients had a cancer at the cardia or fundus vs 44%), the presence of a normal mucosa (3% vs 30%) and atrophy in the antrum (53% vs 17%). Seven of the ten patients with a normal mucosa had a cancer located at the cardia (p < 0.05) and in nine of the eleven patients younger than fifty, the cancer was of the diffuse type (p < 0.005). Thus, patients with H. pylori and gastric cancer differ from those uninfected. Of future concern is the large increase in cancers of the cardia, a cancer unassociated with H. pylori. PMID- 9396173 TI - Omeprazole, nitrendipine, famotidine and stress-induced ulcers. AB - BACKGROUND: Omeprazole inhibits gastric acid secretion by blocking the proton pump of the gastric parietal cell. Nitrendipine is a derivative of the dihydropyridine group of calcium channel blockers and administrated for angina and hypertension. Famotidine is one of the newer histamine H2-receptor antagonists and heals gastric and duodenal ulcers by reducing gastric acid output. OBJECTIVES: The healing effects of omeprazole, nitrendipine and famotidine on stress-induced gastric ulcers were investigated in rats. METHODS: Forty male Wistar-albino rats were separated into five groups (n = 8), a control (non-stress) and four experimental (stress) groups. Experimental rats were treated with omeprazole, nitrendipine, famotidine or a placebo after the stresses of starvation and cold-restraint. RESULT: Macroscopically, the mean area of the affected lesional mucosa was 1/4 of the total gastric mucosa in the famotidine treated group and 1/5 of the total gastric mucosa in the nitrendipine treated group. A considerably decrease was observed in the omeprazole treated group in which the mean area of the lesional mucosa was only in 1/8 of the total gastric mucosa. On microscopic examination, congestive vessels and chronic inflammatory cell infiltrates were significantly reduced in the omeprazole treated group. Tissue regeneration was more prominent in the omeprazole group than the other groups. CONCLUSION: Omeprazole was found to be superior in terms of the effect on the healing process to nitrendipine and famotidine. Although therapeutic effects of nitrendipine and famotidine were observed, those were less than omeprazole. PMID- 9396174 TI - Short- and long-term efficacy of cyclosporin administration in patients with acute severe ulcerative colitis. Belgian IBD Group. AB - Cyclosporin (CsA) has been proposed in the management of patients with acute ulcerative colitis (UC) in whom standard therapy failed and who were candidates for colectomy. Seven academic hospitals contributed to this retrospective study that included 29 patients (median age: 33 y. (15-74 y.); 12 females and 17 males). The median duration of the disease was 4 y. (0.3 to 33 y.). Before initiating CsA, patients were unresponsive to treatment including i.v. corticosteroids (n = 29), 5-ASA or salazopyrine (n = 19), azathioprine (n = 3), antibiotics (n = 14). The i.v. mean dose was 4 mg/kg/day and was adapted to blood level. Concomitant treatment included corticosteroids (n = 27). The median duration of i.v. CsA administration was 10 days (4 to 41 days). At the end of CsA administration, a global improvement was described in 20 patients while a surgery had to be performed immediately in 8 patients because of exacerbation of symptoms (n = 7) or perforation (n = 1). One other patient (74 y.) died because of Pneumocystis carinii infection. For the responders, maintenance therapy included: tapering dose of steroids (n = 12), azathioprine (n = 12), 5-ASA or salazopyrine (n = 10), methotrexate (n = 1) or oral CsA (n = II). The median duration of follow-up was 12 months (4 to 48 months). Among the 20 responders, 7 were subsequently referred for colectomy either electively (n = 3) or because of recurrence of the disease (n = 4). Among the 12 patients treated by azathioprine as a maintenance therapy, only 3 had to be referred for surgery (25%). Among the 8 patients who did not receive azathioprine, 4 were subsequently referred for a colectomy (50%) (NS). In patients with acute refractory UC who received CsA, the short-term efficacy (avoidance of immediate colectomy) was obtained in 20 out of 29 patients (69%). However, after a median follow-up of 12 months, only 13 patients were colectomy free (45%). PMID- 9396175 TI - Genetics and inflammatory bowel disease: from association studies to wide genome screen. PMID- 9396176 TI - Treatment of chronic hepatitis C: practical aspects. AB - The initial enthusiasm about the potential of alpha-interferon therapy for chronic hepatitis C might be weaning in view of the low virus clearance rate of about 15%. At the same time, the potential of interferon to induce sustained remission of the disease is rapidly growing by 3 modalities: a) prolongation of therapy from 6 to 12 months in order to reduce relapse; b) combination of interferon with ribavirin to reduce early non-response, breakthrough and relapse; c) high dose induction therapy; Benefit-risks/cost ratios vary for the different stages of chronic hepatitis C. Consensus exists on the indication of therapy for chronic hepatitis with fibrosis, whereas benefits in very early and very late stages of chronic hepatitis C are doubtful; patients with cirrhosis are clearly in need of therapy but remission is less than 10% and such patients are encouraged to participate in controlled trials assessing combination and/or newer therapy modalities. For decompensated cirrhosis, liver transplantation is the treatment option of choice. PMID- 9396177 TI - Practical management of patients treated with alpha interferon. AB - Interferon alpha is currently used in chronic hepatitis and side effects are well known. They always must be kept in mind to start and to follow a patient under this therapy. A large number of autoantibodies may appear during interferon therapy, usually without clinical manifestations. The detection of dysthyroidism, requires measurement of antithyroid antibodies and TSH before and during interferon therapy. Exacerbation of chronic liver disease under IFN may be found in case of seroconversion in a patient with hepatitis B cirrhosis or in patient with a misdiagnosis of autoimmune hepatitis. Neurolopsychological disturbances are frequently reported; most of them spontaneously disappear. However, depression must be detected because of the risk of attempted or successful suicide. Worsening or sudden onset of psoriasis or lichen planus have been reported in patients treated with interferon. Appearance or aggravation of some clinical symptoms and biochemical tests may threaten life's patient under IFN therapy. The decision to maintain or to interrupt therapy should take into account the response to interferon and the severity of side effect. PMID- 9396178 TI - HCV infection and liver transplantation. AB - Acute and chronic liver diseases related to hepatitis viruses are the main indications for liver transplantation. The risk of viral reinfection after transplantation is the main limitating factor in these indications. HCV reinfection was demonstrated by demonstrating a sequence homology of the hypervariable region of HCV RNA in 2 patients before and after liver transplantation. HCV reinfection is almost constant, assessed by the persistence of HCV RNA in serum in 90% of cases. Acute lobular hepatitis appeared in 75% of patients at a median of 4 months post-transplantation with extremes between 23 days and 4 years. In our series, the 5 year actuarial rate of HCV acute hepatitis on the graft, chronic hepatitis and cirrhosis was 75%, 60% and 8%, respectively. HCV RNA level is dramatically increased after transplantation and seems to correlate with the occurrence of acute hepatitis on the graft. A positive relationship between genotype 1 b and prevalence and severity of HCV hepatitis on the graft have been suggested in European series. There is no demonstrated way to prevent HCV reinfection. The use of interferon for the treatment of HCV hepatitis on the graft was disappointing due to a poor antiviral effect and the occurrence of chronic rejection episodes in some patients. Promising results of the combination of interferon and ribavirin have been reported and need confirmation. The 5 year survival of patients transplanted for viral C cirrhosis in our Center is 78%. In conclusion, patients with endstage HCV cirrhosis are candidates for liver transplantation. Viral C reinfection is frequent, but medium term survival is good. However, longterm graft and patient survival remains unknown, and methods to prevent and treat HCV reinfection on the graft are needed. PMID- 9396179 TI - Ascorbic acid metabolism and cancer in the human stomach. AB - The high levels of ascorbic acid (vitamin C) which are maintained in the gastric mucosa and in normal gastric juice suggest that the vitamin has a metabolic role within the stomach. Epidemiological associations between foods containing vitamin C and a reduced risk of gastric cancer together with the ability of ascorbate to quench the mutagenic activity of reactive species produced in the gastric environment, indicate a potential role in the prevention of carcinogenesis. This short review assess the balance of the current evidence and indicates that gastric ascorbate could provide some protection against the development of gastric cancer. However, it is only depletion of ascorbate from the gastric lumen that is likely to contribute significantly to increased risk of malignancy. PMID- 9396180 TI - The role of nitric oxide in portal hypertensive systemic and portal vascular pathology. AB - Hypotension, low systemic vascular resistance and reduced sensitivity to vasoconstrictor are features of hyperdynamic syndrome in portal hypertension (PH) and are pathogenetic factors triggering most serious clinical complications of liver cirrhosis. Nitric oxide (NO) is a powerful vasodilating agent, released from vascular endothelium cell and effecting relaxation of vascular smooth muscle. An increased release of NO has been proposed to play a role in the pathogenesis of vasodilation and vascular hypocontractility associated with PH. In agreement with this hypothesis, the whole-body production of NO has been found to be increased in PH, and the measurement of NOS mRNA expression in different organs suggest that the splanchnic vascular system is a major source of NO release. Consequently, NO could play a role in the development of the splanchnic hyperaemia, collateral circulation and portal hypertensive gastropathy. Furthermore, increased generation of NO in central circulation likely accounts for pulmonary vasorelaxation and cardiac dysfunction found in cirrhosis. By contrast, PH-associated endothelial dysfunction seems to invalidate the capability of intrahepatic and intrarenal vasculature to produce NO. A deficient NO release in these vascular territories might contribute to enhancement of PH and development of the hepatorenal syndrome. Overall NO hyperproduction is either the cause (induction of iNOS) or the consequence (stimulation of ecNOS) of the hyperdynamic syndrome. This incertitude results from the yet undefined significance of mild and transitory activation of the endotoxin-cytokines axis for iNOS induction and contradictory data on specific iNOS and ecNOS activities. A contribution of each isoform of NOS to pathogenesis of the hyperdynamic syndrome probably depends on the model of PH in animal studies and the aetiology or severity of cirrhosis in human studies. PMID- 9396181 TI - The role of TIPS for the treatment of portal hypertension: effects and efficacy. AB - In patients with variceal bleedings TIPS is effective even if the portal pressure is reduced only partially and the reduction does not reach the threshold of 12 mmHg. Since the post-TIPS pressure gradient is closely correlated to the incidence of hepatic encephalopathy, higher gradients should be favoured in patients with a higher risk of hepatic encephalopathy, e.g. patients > age 65 years, Child-class C patients, and active alcoholics. An 8 mm diameter-shunt is probably the adequate size for most of these patients. Regarding patients with ascites, the effect of TIPS is partially due to an improvement of renal blood flow and function. The reasons for this are unknown. The systemic hemodynamic effects of the TIPS are probably not the cause since the shunt did not result in an improvement of the arterial filling and peripheral resistance. The experimentally proven hepatorenal baro-reflex may be an explanation. PMID- 9396182 TI - Cystic gastric leiomyoma--a diagnostic pitfall. AB - A case of a 74-year-old woman with a cystic calcified leiomyoma of the stomach is presented. The cyst was initially interpreted and treated as a pancreatic pseudocyst with repeated punctures and cystogastrostomy. Due to failure of elimination of the cyst, and because of infection, the patient underwent surgery. A discussion of the differential diagnoses and treatment is presented. PMID- 9396183 TI - Fundic gland polyps: three other case reports suggesting a possible association with acid suppressing therapy. AB - We describe three patients who developed fundic gland polyps after starting a treatment with acid suppressive agents. All three patients were treated for a long time with H2-blockers or proton pump inhibiting agents for reflux oesophagitis. In one patient the fundic gland polyps disappeared after discontinuation of the acid suppressing therapy. A possible causal role of these agents is suggested. A review of the literature about fundic gland polyps and their possible association with acid suppressive therapy and a short review about proton pump inhibiting agents and the appearance of gastric polyps in general is given. PMID- 9396184 TI - Increased transaminases in psychiatry: a case report. AB - We report the case of a patient admitted to the hospital with psychiatric troubles. Soon after admission, he presented severe hepatitis of unknown origin. Careful review of the charts, transvenous liver biopsy, right heart and hepatic pressure measurements, negative toxicologic and viral screenings were highly suggestive of hypoxic hepatitis. Indeed, the patient had previously been treated for a decompensated cardiomyopathy and medications stopped prior to the current admission. Without clear clinical evidence of heart failure he presented a brief malaise two days before the increase in liver enzymes. Holter heart recording showed afterwards bouts of ventricular tachycardia. Treatment with Dobutamine and antiarrythmics led to a rapid decrease of transaminase levels and recovery in liver function. Unfortunately, he died three weeks later from his cardiomyopathy. This case illustrates the need for cardiovascular work-up in the context of hepatitis from unknown origin. PMID- 9396185 TI - Preparing and interpreting meta-analysis in clinical research. AB - Meta-analysis is a procedure to systematically research and collect published and (ideally) unpublished randomized clinical trials (RCTs) of treatments and quantitatively summarise their results, in order to obtain an objective assessment of efficacy. Collaboration between statisticians and clinical experts in the field of pathology addressed by the treatment is needed for performing reliable meta-analyses. "Typical" meta-analysis can be refined as "cumulative" meta-analysis, where the pooled assessment of treatment effect is repeated every time a new trial is added to a set of trials, and by meta-analysis "on individual patient data", i.e. using information on each patient included in every trial. Due to the emergence of Evidence-Based Medicine (EBM), the role of meta-analysis is expanding. EBM is a way of practising medicine by integrating individual clinical expertise with the most reliable evidence from the medical literature. Meta-analysis is suggested as the most convenient and reliable source of data for practising EBM. EBM is taking advantage from the Cochrane Collaboration, an international network of experts performing, updating and disseminating meta analyses of important treatments, according to a common model and established procedures. PMID- 9396186 TI - Molecular biology in uro-oncology: clinical application. Prognostic factors in bladder cancer. AB - In the bladder cancer the most important prognostic factors are the stage, the grade, the presence or absence of lymph nodal metastasis, the response to therapy with B. C. G. etc.... In any case, even in the context of the same clinical stage, it is not possible to correctly evaluate the evolution of the disease. The Author did a literature revision and got a personal contribution about the effective utility of same biological prognostic factors. In a study about superficial bladder tumor using monoclonal antibody MIB-1 (Ki-67) a correlation between proliferation index (P.I.) and grade was noted. In particular the presence of a P.I. above 40% correlated with greater precocity and frequency of recurrences. A similar study showed that the expression of protein p21 correlated with a greater precociousness and with recurrence frequency. In conclusion, we have also carried out an evaluative study on the expression of oncosuppressor gene p53. In superficial bladder cancer this study showed up a correlation between the expression of protein p53 and a greater precociousness and frequency of recurrences. PMID- 9396188 TI - Molecular genetics of renal cell carcinoma. AB - Significant research progress over the last few years has identified several major genetics contributors to RCC. A new classification of RCC validated by cytogenetic and molecular studies has been proposed including nonpapillary, papillary, chromophobe and oncocytic tumors. The cytogenetic analysis of patients with familial RCC, VHL disease and sporadic RCC have shown that WHL gene located on chromosome 3P 25 is a tumor suppressor gene. Other genes may be involved in the development of RCC, however with a less important incidence than VHL gene. Mutations of Rb and P53 genes can be associated with metastatic disease, mutations of the ras gene is rare whereas elevated level of myc oncogene are frequent but of little prognostic value. Controversial the role of ploidy and proliferation markers as independent prognostic factors. PMID- 9396187 TI - Effects of LHRH agonists on the growth of human prostatic tumor cells: "in vitro" and "in vivo" studies. AB - Luteinizing Hormone Releasing Hormone (LHRH) agonists exert both "in vitro" and "in vivo" a direct inhibitory action on the growth of both androgen-dependent (LNCaP) and androgen-independent (DU 145) human prostatic cancer cell lines. The present experiments have been performed to investigate the mechanisms involved in this direct antiproliferative action of LHRH agonists. In particular, the aim was to study whether these compounds might exert their antiproliferative effect by interfering with the stimulatory action of epidermal growth factor (EGF) both "in vitro" and "in vivo". To this purpose, the effects of LHRH agonist, Zoladex (LHRH A), on the mitogenic action of EGF, on EGF-activated intracellular signaling mechanisms (tyrosine phosphorylation of EGF receptor and c-fos proto-oncogene expression), and on the concentration of EGF receptors have been evaluated in both LNCaP and DU 145 cells. The results of these "in vitro" studies show that in LNCaP cells LHRH-A counteracts the mitogenic action of EGF, abrogates the EGF induced c-fos expression and reduces the concentration of EGF-binding sites, without modifying the EGF induced tyrosine phosphorylation. In DU 145 cells, LHRH A antagonizes the proliferative action of EGF, inhibits tyrosine phosphorylation of EGF receptor induced by EGF and significantly reduces the number of EGF binding sites, without altering the stimulation of c-fos expression induced by EGF. For the "in vivo" experiments, male nude mice were s.c. injected in the flank with DU 145 cells and treated for 14 days with LHRH-A (100 micrograms/days). At the end of the treatment, the concentration of EGF receptors on membrane preparations as well as on tumor volume were found to be significantly lower in LHRH-A treated animals than in control mice. The mitotic index and the expression of the proliferation-associated antigen Ki67 were found similar in control as well as in treated animals. In addition no modification of apoptotic index (expression of p53) was observed. These data suggest that LHRH agonists may inhibit the proliferation of the tumor cells by interfering with the stimulatory actions of EGF. PMID- 9396189 TI - Simple renal cysts. AB - OBJECTIVES: To evaluate the incidence of simple renal cyst, the relationship between both size and location of cysts, and the effect on calyces; and to correlate symptomatology and effect on the pelvicalyceal system. METHODS: Abdominal ultrasound examination was performed in 2010 patients for different reasons. Simple renal cysts were demonstrated in 110 patients harbouring 198 cysts. The cysts were divided into 3 categories according to their size. The relation between the size of the cyst to the clinical symptoms and to the effect on the pelvicalyceal system is discussed. The clinical presentation in relation to effect on calyces is evaluated. RESULTS: The male to female ratio was 1.4:1. The majority of the simple renal cysts were encountered in patients above the age of 50 years. However, the incidence of renal cysts in children in this series was 2% which is considered high as compared to other studies. Thick wall, marginal calcification of cysts and multilocularity, each was seen in 1% of the cases. Associated liver cysts was seen in 2% of the cases. CONCLUSIONS: We can conclude that there is an important relationship between both size and location of cysts, and the effect on calyces; however, no noticeable correlation between symptomatology and effect on the pelvicalyceal system was observed. PMID- 9396190 TI - Clinicopathologic correlation in a case of metastatic uveal tumor. AB - A clinicopathologic correlation is reported of an ocular metastasis from an unknown primary tumor. The tumor appeared initially confined to the choroid. The diagnosis of metastatic adenocarcinoma was obtained by choroidal biopsy. The metastasis was uncontrollable with teleradiotherapy. Six months later the anterior segment also appeared infiltrated and the eye was enucleated. PMID- 9396191 TI - Indocyanine green angiography versus fluorescein angiography in the follow-up of choroidal melanomas treated with RU106/RH106. AB - Indocyanine-green angiography and fluorescein angiography may complement each other in the follow-up and evaluation of choroidal melanomas treated with brachytherapy and may give a better understanding in the process of response of choroidal melanomas to brachytherapie. We did a retrospective study on 18 patients treated for this pathology. PMID- 9396192 TI - Ocular burn caused by soft brown soap. AB - PURPOSE: To report the danger of serious chemical ocular burn caused by common household soap. CASE HISTORY: A 20 year old male developed an extensive burn of the right eye after the commonly used soft brown soap (also called floor soap) fell into his face and right eye. The burn caused conjunctival ischemia and necrosis. The cornea was oedematous and denuded of epithelium. The pH of the soap was 11.8. The patient received treatment for alkali burn of the eye. Stem cell transplantation was needed to heal the corneal surface defect. Scar formation and peripheral neovascularisation have reduced the visual acuity to counting fingers at 75 cm. CONCLUSION: This case of serious alkali burn caused by the common soft brown soap demonstrates the potential hazard to the eyes. The manufacturer was sought to write the pH and a warning on the exterior of the container that the "soft brown soap" may cause serious ocular injury. PMID- 9396194 TI - Primary acquired melanosis and melanoma of the conjunctiva. AB - Primary acquired conjunctival melanosis presents as a unilateral conjunctival pigmentation, mostly in middle-aged patients, with a strong tendency to progress to malignancy. The clinical picture is specific and doesn't cause major diagnostic problems. It is important to recognize the entity, to observe it closely and treat it early and adequately. PMID- 9396193 TI - Pelli-Robson contrast sensitivity test in Zaire. AB - The purpose of this study was to establish in this first report the age standards of the Pelli-Robson contrast sensitivity in Zaire. Contrast sensitivity using the Pelli-Robson chart was performed in 100 normal Zairian black subjects aged from 10 to 59 years and 36 patients (22 patients with open-angle glaucoma and 14 with optic nerve disease). Scores of normal subjects were age related (p < 0.05). The results of Zairian young subjects were similar to those found previously in young white subjects; scores for older subjects were lower when compared to those of whites. Scores of patients were lower than those of normals (p < 0.001). Contrast sensitivity using the Pelli-Robson chart can be useful in developing countries. PMID- 9396195 TI - Sneddon's syndrome in a patient with homonymous hemianopia with macular sparing. AB - A 48-year-old women is described with the infrequent association of generalized livedo reticularis and cerebrovascular accident of idiopathic origin (Sneddon's syndrome-SS). Visual field testing revealed a left homonymous hemianopia with macular sparing. Though visual field impairments in SS have been reported, the type could usually not be specified precisely because of the dementia and lack of cooperation of the patients. PMID- 9396196 TI - I-123-IDAB: a new tracer for scintigraphic visualisation of malignant melanoma. AB - I-123-N-(2-Diethylaminoethyl) 4-lodobenzamide (I-123IDAB) has recently been introduced as a radiopharmaceutical agent in the management of patients with malignant melanoma. We tested the diagnostic potential of this substance in 5 patients suspected of having an ocular malignant melanoma. I-123-IDAB scintigraphy identified 4 patients with high tracer uptake in the pathologic eye. Three cases were confirmed histologic as a malignant melanoma, the fourth patient was submitted to radiotherapy. The amelanotic (histologically confirmed) irismelanoma of the fifth patient could not be detected. From these preliminary results we conclude that I-123-IDAB scintigraphy may be a valuable tool for establishing the diagnosis of malignant melanotic melanoma. PMID- 9396197 TI - Indocyanine green angiographic findings in multifocal choroidopathies. AB - With high definition videoangiography (TOPCON IMAGEnet H1024) the Authors studied 41 patients affected by multifocal choroidopathies (MC) (68 eyes with ophthalmoscopic or indocyanine green angiographic evidences): 29 females and 12 males; age 21-51 years with a follow up of 6-29 months. In the light of the evidence provided by FA and ICG the Authors present a classification of MC in three stages: Stage 1 of subclinical choroidal activity (5 eyes): characterised by the presence of hypofluorescent or hyperfluorescent spots visible only in the late phases of ICGA; stage 2 of clinically evident choroidal activity (45 eyes): in FA the spots are hypofluorescent in the early phases and hyperfluorescent with a slight diffusion in the late phases, in ICGA either hypofluorescent spots or less frequently hyperfluorescent spots and choroidal permeability alterations can be observed; stage 3 or healed stage (18 eyes): in FA the spots are hyperfluorescent without late leakage, in ICGA hypofluorescence can be observed during all angiographic phases. In 5 patients in stage 1 of subclinical activity, a systemic steroid therapy induced the regression of the hypofluorescent sports in ICGA, in 2 cases the regression of hyperfluorescent spots in ICGA was observed after systemic antibiotic therapy. The authors underline that ICGA could be a particularly useful tool for an early diagnosis and clinical monitoring of MC. PMID- 9396198 TI - Steps toward discovering causes: divergence and convergence of epidemiology and clinical medicine. AB - Ways in which frames of reference about causation differ across disciplines are exemplified. The necessity for a multivariate concept of causation is emphasized. On the epidemiological side, divergences of clinical approaches from requirements for meeting preconditions for cause are each illustrated by several examples. Association is discussed in terms of comparability of groups, independent observation of associated variables, sampling and measurement. Securing time order among variables is a matter of sequential positioning by design, and the difficulties that inhere in the clinical situation are again illustrated by example. On the clinical side, strengths in generating hypotheses, in discovery of associations without conventional comparison and in potential precision in measuring outcomes are outlined. On the epidemiological side, the discipline adds to clinical medicine unique dimensions analogous with the basic biomedical sciences. These are essential to capturing environmental causes and the antecedent experience of sick individuals. Convergence between the two perspectives is bound to yield, and has yielded, much profit. PMID- 9396199 TI - Geriatric medicine. A new discipline for the coming century. PMID- 9396200 TI - Cardiovascular disease in the elderly. PMID- 9396201 TI - Isolated systolic hypertension in the elderly. PMID- 9396202 TI - Kidney diseases in the elderly. PMID- 9396203 TI - Infections in the elderly. PMID- 9396204 TI - Cancer in the elderly. A special and unique entity. PMID- 9396205 TI - Intracranial sewing needles in a 20-year-old patient. AB - A 20-year-old patient is reported with three intracranial sewing needles, which were located in the frontal lobes. The clinicoradiologic findings strongly suggested that they must have been placed into the brain through the anterior fontanelle during an unsuccessful homicide attempt in infancy. PMID- 9396206 TI - Paraganglioma of the cauda equina: MR findings. One case. AB - Paraganglioma of the filum terminale is a rare tumor but well described in the neurosurgery and pathology literature. Few MRI reports are mentioned. Paraganglioma, often misdiagnosed with ependymoma or schwannoma on MRI images, must be kept in mind, when a highly vascular lesion with serpentine vessels is observed. PMID- 9396207 TI - [One hundred years of sinusoidal cells in the liver]. AB - Kupffer (1898) reported that the stellate cells were phagocytic endothelial cells, retracting his earlier view that these cells were perivascular cells. His new concept stimulated studies on the vital staining and the RES theory, while it ensued several controversies in liver histology. The original stellate cells were rediscovered in 1971, and were proved to be identical with several perisinusoidal cells reported previously. For this cell the term hepatic stellate cell has recently been adopted as the standardized name. The space of Disse is newly defined as the space between the endothelial cell-stellate cell complex and the parenchymal cells. The stellate cells display vitamin A-storage, collagen synthesis and may contract to regulate the sinusoidal blood flow and the fluid exchange between the sinusoidal lumen and the space of Disse. The sinusoidal endothelial cells uptake injected lithium carmine and macromolecules by coated vesicles. Kupffer cells clear endotoxin, bacteria and apoptotic neutrophils and release various cytokines. The pit cells are the liver-associated NK cells and are activated by the administration of biological response modifiers, preventing tumor metastasis. Extrathymic pathways of T cell differentiation exist in the hepatic sinusoid. The dendritic cells differentiate in the sinusoid and translocate to the Glisson's sheath. Intralobular heterogeneity of sinusoidal cells have been observed. The sinusoidal cells communicate each other with autocrine and paracrine mechanisms to regulate various functions of the liver. PMID- 9396208 TI - [Historical notes on anatomy of the transversalis fascia]. AB - In the early 19th century, the tissue of the peritoneum was regarded as the duplicature of membrane. In the middle of the century, however, it was considered to be one of serous membranes, because histology had been developed. Between the peritoneum and the abdominal muscles, there are two fasciae, the subperitoneal and the transversalis fascia. But, Sir Astley Cooper reported that there was only the transversalis fascia, because he considered the subperitoneal fascia to be a double covering of the peritoneum, that is, a part of the duplicature of the peritoneum. In this century, Cooper's report has been interpreted through histology. Consequently, it is the general opinion that there is no other fascia than the transversalis fascia between the peritoneum and the abdominal muscles, in a view which disregards the subperitoneal fascia. PMID- 9396209 TI - [Integrins: their structures, functions and gene expressions in the central nervous system. Acta anat]. AB - More than ten years have passed since integrin was shown to function in cellular attachment. To date integrin research has been one of major fields in cell biology. Integrin, which functions as an integrator of both extra- and intracellular skeletal molecules, is regarded as one of the essential molecules for cellular signal transduction as well. Thus, integrin appears to be essential and indispensable for many cellular phenomena. Although every type of cell is thought to express a few kinds of integrin molecules, their expression and functional roles in neurons remain to be determined. Both intensive and extensive researches should reveal one by one how integrins are involved in the neural network formation in development, neuronal plasticity and regeneration, higher function of CNS, and also neuronal degeneration in both inflammation and degenerative diseases. PMID- 9396210 TI - Differential accumulation of calcium and phosphorus in aged human arteries. AB - To elucidate age-related changes of human arteries, relative contents (RCs) of minerals were analyzed by inductively coupled plasma atomic emission spectrometry on the thoracic aorta, basilar, coronary, femoral, and radial arteries from 27 subjects within the age range between 0 and 92 years old (Ys). Calcium and phosphorus never accumulated uniformly in any of the arteries such as the thoracic aorta, basilar, coronary, femoral, and radial arteries. The accumulations of calcium and phosphorus occurred earlier in the order of the femoral artery, thoracic aorta, coronary artery, and basilar or radial artery. PMID- 9396211 TI - Right superior bronchial artery arising from the right subclavian artery and accompanying nerve branches: an autopsy case. AB - In one out of 8 examined cadavers a solid, right superior bronchial artery was found to arise from the subclavian artery with its origin at the same level as those of the right vertebral and internal thoracic arteries. This bronchial artery was 1.5 mm in caliber and closely associated with the right sinal nerve arising from the vagus nerve and with the right stellate cardiopulmonary nerves arising from the right stellate ganglion. Such a close association was also observed between other sympathetic cardiac nerves and bronchial arteries and an extracoronary artery. On the basis of these observations it was deduced that the general course of the bronchial arteries serves to pave the way for the extension of the sympathetic cardiac nerves to the heart. PMID- 9396212 TI - Trends in cancer mortality: perspectives from Italy and the United States. AB - Cancer continues to be a major public health problem in Italy, as it is throughout the world. We analyzed age-adjusted mortality rates for all cancers combined and for specific cancer sites in Italy for five year periods from 1950 to 1989. We compared trends in Italian cancer mortality to those observed in the United States during the same time period. We also considered some ancillary data including age-specific mortality rates, as well as incidence and five year relative survival data from the Modena Province. Age-adjusted cancer mortality rates in Italy are increasing in males and, to a lesser extent, females. This finding is in contrast to a recent plateau in age-adjusted cancer mortality rates in the United States. In Italy, stomach cancer mortality has declined substantially, counteracting marked increases in lung cancer mortality, particularly in males, and breast and lung cancer in females. Changes in cancer mortality in Italy, as in the US, have been driven primarily by changes in disease incidence rather than advances in therapeutics. These data suggest a need for realignment of cancer control resources toward prevention, particularly with regard to lung cancer and tobacco usage. PMID- 9396213 TI - Introduction: history of the use of asbestos. AB - Discussion of the major milestones in the history of the modern uses of asbestos and the first knowledge of the health effects associated with such usage. Highlights of the studies associating exposure to asbestos with non-malignant lung diseases, lung cancer, and mesothelioma are discussed. PMID- 9396214 TI - Asbestos-related mortality in Italy: a geographical approach. AB - The present contribution describes two studies of asbestos-related cancer mortality in Italy: an analysis of the geographical distribution of mortality from pleural neoplasms and an investigation into the relationship between pleural and lung cancer mortality in an Italian region, Piedmont. Mortality from malignant pleural neoplasms (ICD-IX Revision 163.0-163.9) has been studied in Italy in the 20 regions and the over 8000 municipalities for the years 1988-92: restriction of analysis to municipalities with at least three observed deaths and statistically significant increases led to identification of areas where occupational and/or environmental exposure to asbestos can have occurred. The analysis of pleural and lung cancer mortality was carried out for the years 1980 87 in Piedmont using an empirical Bayes method for small-area disease mapping; the results showed that the proportion of lung cancer mortality attributable to asbestos exposure was 5.6 and 5.7 percent respectively in men and women. While analyses of routinely collected data are no substitute for ad hoc individual based studies, notwithstanding the limitations of geographical approaches to the study of asbestos-related mortality, investigations carried out at the level of small area populations appear to provide informative results, which might be of value in terms of public health. PMID- 9396215 TI - Pleural malignant mesothelioma and environmental asbestos exposure in Casale Monferrato, Piedmont. Preliminary analysis of a case-control study. AB - A case-control study on pleural malignant mesothelioma (MM) was conducted in Casale Monferrato, where the largest Italian asbestos cement (AC) factory had been operating from 1907 to 1985. In a previous study we observed a five to seven fold increase in the incidence of MM among people living in that city and never employed in the factory mentioned. The present study includes cases of MM with histological diagnosis over the period 1.1.1987-30.6.1993 among residents in the Local Health Unit (LHU) of Casale Monferrato. Population controls were randomly extracted from the list of the residents in the LHU, matched to cases on sex, date of birth, vital status and date of death. Cases and controls (or their closest relative) were interviewed with a standardised questionnaire focusing on asbestos exposure in the (life-long) residential and occupational histories and in leisure time activities as well as on occupational asbestos exposure of relatives and cohabitants, smoking and chest or occupational diseases. The interview was blind in respect to case or control status. For the analyses the addresses were coded on map grids with a 500 m. mesh size. Statistical analyses were conducted with conditional logistic regression in order to keep the matching between cases and controls. Eighty-eight cases and 244 controls were interviewed (95.6% of cases and 80.1% of controls): 26 and 11 respectively reported an activity in the AC industry. Seven cases and 7 controls were also exposed because of parental occupation. The main analyses are based on the conditional regression model including both occupational and residential exposure. The different modes of exposure are included on an ordinal scale: each subject is classified according to their highest level. Domestic exposure is included as an independent factor. Odds Ratios (OR) are estimated with reference to subjects without either occupational or residential exposure. The OR is 39.3 among subjects reporting occupational exposure, 11.9 among those never engaged in the AC industry but living within 1000 m. of the factory. A statistically significant risk is also observed for persons at some time living in the other areas of Casale Monferrato. PMID- 9396217 TI - Malignant mesothelioma of the pleura in the Trieste-Monfalcone area, with particular regard to shipyard workers. AB - A series of 421 malignant pleural mesotheliomas, diagnosed in the Trieste Monfalcone area, northeastern Italy, were reviewed. A large majority of the patients had been employed in "naval" work (shipbuilding, maritime trades, and dock work). Latency periods (time intervals between first exposure to asbestos and death), showed wide variations from one occupational category to another. Such variations were attributable, but only partly, to differences in the intensity of the exposure to asbestos. Various family cases were identified, including people with and without blood relationships. The data, obtained in the studies on Trieste-Monfalcone mesothelioma, suggest that interactions between asbestos and other factors play a considerable role in the pathogenesis of asbestos-related mesothelioma. PMID- 9396216 TI - The experience of the Mesothelioma Registry of Tuscany in assessing health hazard associated with asbestos exposure. AB - All cases of pleural malignant mesothelioma occurring in Tuscany were collected, backdated to 1980 (to 1970 for the provinces of Florence and Prato; to 1975 for the provinces of Pisa and Siena), in order to evaluate the incidence of occupational exposure to asbestos. The aim was to enhance primary prevention in those workplaces still at risk nowadays. To achieve information on the possible occupational, domestic or environmental exposure, an interview was conducted using a semi-structured questionnaire. An exposure classification was produced to focus preventive intervention. This surveillance system needs to be developed to contribute to epidemiological research, especially on the effects of low level exposures, and to primary prevention. PMID- 9396218 TI - Mesotheliomas in some selected Italian population groups. AB - The analysis of 335 cases of mesothelioma observed at the Ramazzini Foundation and the Bologna Institute of Oncology has shown: 1) a high percentage of correlation of these tumours with asbestos exposure; 2) a large number of population categories potentially exposed to asbestos fibres and therefore at risk of developing mesothelioma; and 3) the high risk of mesothelioma among people exposed in various circumstances to asbestos used in railroads and sugar refinery plants. PMID- 9396219 TI - The riddle of risk assessment in asbestos carcinogenicity. AB - While there appear to be some legitimate issues of scientific inquiry with regard to asbestos carcinogenicity, there also appear to be continuing areas of controversy which, given the state of scientific evidence, should be considered settled. There have been real costs, both monetarily and in human suffering, with regard to past and current uses of asbestos. With the continuing, often poorly controlled exposure, that workers still have, more disease can be expected. The fitting remembrance of Dr. Selikoff would be to remember the science that he contributed and the interest he took in the lives of workers and others. It then becomes incumbent upon those of us who continue to work in this area to carry on in the spirit of protecting the lives and well-being of workers from this hazardous material, as we would for other known hazards. PMID- 9396220 TI - Conclusion: prevention of asbestos-induced disease. PMID- 9396221 TI - DNA flow cytometry and 67Ki proliferating index as prognostic factors of early recurrence and progression in G1-G2/Ta-T1 and G3/Ta-T1 transitional cell carcinoma of the bladder. AB - Using flow cytometry, gross genomic alterations, defined as DNA ploidy, and the fraction of S-phase cells, can be calculated in bladder cancer cells. In aneuploid superficial bladder cancer the recurrence rate has been reported to be three times higher than in diploid forms. A correlation between the S-phase fraction and progression has been reported for G1-G2/Ta-T1 tumours, but not for G3/Ta-T1. The aim of our study is to evaluate whether the traditional cytometric parameters can be used as valid predictors of early recurrences and progression in G1-G2/Ta-T1 and G3/Ta-T1 bladder cancer patients and to compare the proliferation indexes as defined by S-phase fraction and 67Ki monoclonal antibody in the two groups of patients. PMID- 9396222 TI - Sarcomas of the kidney. AB - Sarcomas of the kidney are exceedingly uncommon. They represent between 1 and 3% of all malignant renal tumors. Several histological types have been reported. We have reviewed the renal tumors diagnosed in our department in the last 25 years and found 4 cases of sarcoma out of 390 malignant renal tumors (1.02%). They were a liposarcoma, a storiform-pleomorphic fibrous malignant histiocytoma, a rhabdomyosarcoma and a pleomorphic leiomyosarcoma. We report the clinical data and the outcome of the patients, together with the immunohistochemical and ultrastructural findings in these cases. PMID- 9396223 TI - Villous adenoma of the bladder. AB - Villous adenomas of the bladder are rare tumors and up to now they have not been seen to undergo malignant transformation. We report a case of villous adenoma of the bladder with areas of adenocarcinoma in a 72-year-old man. We describe all the morphological, histochemical and immunohistochemical features characterizing this tumor. We recommend adequate pathological sampling and a thorough follow-up of patients with villous adenoma. The prognosis and the behaviour of these adenomatous papillary lesions, morphologically similar to colonic adenomas, in the bladder is unclear. We report a case with focal area of adenocarcinoma and review the literature. PMID- 9396224 TI - [Acetaldehyde adducts in the cerebral cortex of ethanol-fed mice]. AB - To investigate the neurotoxicity of acetaldehyde covalent adducts, immunohistochemical staining for acetaldehyde adducts using the antibody against acetaldehyde adducts, was performed in the cerebral cortex of ethanol-fed (withdrawal) mice. In the ethanol-fed mice, the degeneration in the cerebral cortex was found, while the protein epitope related to acetaldehyde was found in the cerebral cortex, liver and adrenal cortex. No histochemical and immunohistochemical changes in the tissues from the control mice were found. It is possible that acetaldehyde adducts may effect on the cerebral cortex as the neurotoxicity which cause psychosis such as delirium and hallucination after alcohol drinking. PMID- 9396225 TI - [Mental and physical symptoms in alcoholics after alcohol withdrawal]. AB - Much of the recent interest in the risk factors of relapse in alcoholics has focused on the withdrawal depressive state. Although the clinical relevance of this syndrome is often acknowledged, definite empirical research on subjective symptoms has not been presented in detail. Therefore, the purpose of this paper is to explore the characteristics of mentally or physically subjective symptoms of alcoholics and to discuss the risk factors to "slip". To evaluate the subjective symptoms, the health questionnaire that is composed of 62 items was applied to 73 admitted alcoholics. The principal component analysis obtained for the questionnaire of mental symptoms extracted five factors as followed, 1) feeling of estrangement or emptiness, 2) anxiety or impatient, 3) depressive mood or sleeping disturbance, 4) endogenous depressive factor, 5) unstable emotion. Additionally, five factors on physical symptoms were extracted as follows, 1) autonomic dysregulation related to neurasthenic state, 2) autonomic dysregulation related to hysterical neurosis, 3) loss of libido, 4) appetite loss, 5) dry mouth or sweating. These findings emphasized that many alcoholics felt the feeling of maladjustment and emotionally unstable under the "sobriety" state. These also suggest the "slip" of alcoholics should depend on expectation to reduce their tension, conflict and frustration under the influence of alcohol. PMID- 9396226 TI - [Effect of nicotine-administration on tyrosine hydroxylase-containing neuron in the rat forebrain; immunohistochemical study with semiquantitative morphometric analysis]. AB - Nicotine is toxic substance that is absorbed by cigarette smoking and easily causes dependence. It is clear that smoking is bad for health, however, the effects of nicotine itself on the central nervous system have not been elucidated. We studied the effects of nicotine-administration (5 mg/kg x 2/day, 7 days) on immunoreactivity for tyrosine hydroxylase (TH), the rate-limiting enzyme of catecholamine synthesis, in the rat forebrain including the cerebral cortex, hippocampus, striatum and hypothalamus to investigate the influence on catecholaminergic neurons. In the nicotine-administration group, both the number and intensity of TH-immunoreactive fibers and terminals increased in the fronto parietal cortex, anterior cingulate cortex and hippocampus in comparison with those of the control group, and a significant difference was demonstrated by computer assisted morphometric analysis. These findings suggest that nicotine administration influences catecholaminergic neural systems in the forebrain, especially in the cerebral cortex and hippocampus and that these effects might be related to developing the dependence on smoking. PMID- 9396227 TI - [The ophthalmological application of functional brain imaging]. PMID- 9396228 TI - [Physical properties of an intraocular lens coated with diamond-like carbon film]. AB - I have invented an intraocular lens (IOL) coated with diamond-like carbon film and have investigated its physical properties. Diamond-like carbon film is an ultrathin film which is composed of many carbon atoms and very few hydrogen atoms, and has properties very similar to diamond. I have measured the following properties of the polymethylmethacrylate (PMMA) plates or IOLs coated with diamond-like carbon film: light transmission curve, resistance against Nd:YAG laser capsulotomy, peel-off test, and contact angle. The plates or lenses are more ultraviolet-absorbing than uncoated PMMA plates, and PMMA plates coated with 50 nm-thick diamond-like carbon film have a light transmission curve very similar to that of a 25-year-old human crystalline lens. The pits and cracks due to Nd:YAG laser emission were smaller in the coated IOLs were more hydrophobic and oleophilic than uncoated IOLs. Diamond-like carbon film coated IOLs are thought to be very promising. PMID- 9396230 TI - [Selective binding of fucose-recognizing lectin on the cone photoreceptor outer segments]. AB - The distribution of fucose-containing glycoconjugates in the photoreceptor cell layer of rat and human retinas was examined by lectin histochemistry using Aleuria aurantia lectin (AAL), which recognizes L-fucose alpha 1, 6 residue. In the rate retina, AAL diffusely bound to the apical outer segments and to the basal inner segments, whereas it bound to the entire outer segments of other photoreceptors, which were considered to be cones due to their proportion. In the human retina, AAL bound diffusely to the basal inner segments and to the retinal pigment epithelia, but it bound selectively to the outer segments of the cones. The present findings revealed that the glycoconjugates, whose sugar chains contain L-fucose alpha 1, 6 residue on their termini, are present in the cone outer segments. PMID- 9396229 TI - [Inhibition of experimental autoimmune uveoretinitis by transforming growth factor-beta 1 in B10. A mice]. AB - Experimental autoimmune uveoretinitis (EAU) in mice, an organ specific autoimmune disease, has been investigated as an animal model for human endogenous uveitis. In this study, we report on the immunosuppressive effect of transforming growth factor-beta 1 (TGF-beta 1) on the development of EAU in mice. Inhibition by TGF beta 1 of proliferation of interphotoreceptor retinoid-binding protein (IRBP) specific T cell lines in B10.A mice against IRBP antigen was dose-dependent. However, when spleen cells used as the antigen presenting cell were first cultured with TGF-beta 1, this anti-proliferation effect was abolished. When IRBP immunized mice were injected intraperitoneally with TGF-beta 1, dose-dependent suppression of EAU was obtained. The proliferation response of lymph node cells from TGF-beta 1 injected mice with IRBP-induced EAU was suppressed compared with phosphate buffered saline (PBS)-injected mice. These findings suggest that TGF beta 1 may be a cytokine that plays a role in suppressing IRBP induced EAU in mice. PMID- 9396231 TI - [Results of vitreous surgery for proliferative vitreoretinopathy]. AB - During the past four years, we performed vitreous surgery on 73 eyes with rhegmatogenous retinal detachments complicated with severe proliferative vitreoretinopathy (PVR). We analyzed the surgical outcome of PVR according to the revised classification of PVR Grade C (1991). After a mean follow-up period of 19 months, the retinas were successfully reattached in 62 of 73 eyes (85%). The reattachment rate in the eyes with only posterior proliferation was high (96%), regardless of the extent of posterior proliferation. However, the reattachment rate in the eyes associated with anterior proliferation was markedly low (57%), depending on the extent of anterior proliferation. Among 62 eyes with successfully reattached retinas, 39 eyes (63%) had an improved postoperative visual acuity. These results demonstrated that the eyes with anterior PVR have a worse reattachment rate than the eyes with only posterior PVR. Using the revised classification of PVR, we were able to analyze the surgical outcome of PVR which could not be classified by the old classification. PMID- 9396232 TI - [Long-term visual recovery after scleral buckling of macula-off retinal detachments]. AB - We retrospectively investigated long-term visual recovery in 32 macular reattached eyes which had been followed up for more than five years after surgery. In 17 eyes (53%), the best-corrected visual acuity at 5 years after surgery was > or = 2 lines better than best-corrected visual acuity at 3 months postoperatively. For these 17 eyes, the mean visual acuity continued to improve for up to 10 years after surgery. In the other 15 eyes, visual acuity changes were within 1 line. Improvement of long-term postoperative visual acuity was found to be statistically correlated with younger age, no or mild myopia (< -5D) and shorter duration of macular detachment (< or = 30 days). Surgeons should be aware that the visual function of reattached retinas may recover in the long term, especially in eyes with these features. PMID- 9396233 TI - [Immunohistochemical localization of MUC 1 and keratin 14 in the invasive regions of malignant eyelid tumors]. AB - The distributional patterns of MUC 1 (the mucin whose cDNA was first cloned) and Keratin 14 (K14) in the invasive regions of malignant eyelid tumors were immunohistochemically examined by comparing with other histochemical markers. The MUC 1-positive tumor cells were detected in several serial, small, invasive tumor masses in the deep subepithelial region of the low differentiated carcinoma. They were also continuously detected in the border region between accumulated lymphocytes including T cells and tumor masses of the sebaceous carcinoma. On the other hand, K14-positive tumor cells were detected in the marginal regions of large tumor masses or those with smooth edges, some of which overlapped the distribution of MUC 1-positive cells in the tissues of undifferentiated carcinoma, squamous cell carcinoma, and sebaceous carcinoma. In general, MUC 1 may be expressed in the invasive tumor cells, whereas K14 may be expressed in the marginal cells of the stable, proliferating tumor masses. PMID- 9396234 TI - [The long-term cystic bleb appearance and safety after trabeculectomy with mitomycin C]. AB - The clinical appearance of cystic blebs and the incidence of bleb infection were retrospectively evaluated in 215 trabeculectomies with mitomycin C. The incidence of cystic bleb formation in trabeculectomy was 79% (169/215). The cumulative incidence of the cystic bleb survival when using the Kaplan-Meier method was 73% in the 50th month after surgery. The incidence of large cystic bleb survival was better than that of small and localized cystic blebs. There was statistically significant difference. The incidence of cystic bleb survival without Seidel phenomenon and bleb infection was 96% in the first year after surgery and 91% in the third year. Bleb infections occurred in two of 169 eyes (1.2%). Bleb infection was successfully treated and there were no cases of endophthalmitis. The large cystic blebs had a higher incidence of bleb infection than the small ones. In one of 25 eyes treated with antibiotic eye drops after surgery, the Seidel test was positive and bleb infection occurred. The Seidel test was positive in 5 of 96 eyes in which antibiotic eye drops were not used and bleb infection occurred in one of these 5 eyes. PMID- 9396235 TI - [Detection of cortical activities on eye movement using functional magnetic resonance imaging]. AB - Cortical activity during eye movement was examined with functional magnetic resonance imaging. Horizontal saccadic eye movements and smooth pursuit eye movements were elicited in normal subjects. Activity in the frontal eye field was found during both saccadic and smooth pursuit eye movements at the posterior margin of the middle frontal gyrus and in parts of the precentral sulcus and precentral gyrus bordering the middle frontal gyrus (Brodmann's areas 8, 6, and 9). In addition, activity in the parietal eye field was found in the deep, upper margin of the angular gyrus and of the supramarginal gyrus (Brodmann's areas 39 and 40) during saccadic eye movement. Activity of V5 was found at the intersection of the ascending limb of the inferior temporal sulcus and the lateral occipital sulcus during smooth pursuit eye movement. Our results suggest that functional magnetic resonance imaging is useful for detecting cortical activity during eye movement. PMID- 9396236 TI - [Effects of onpi-to (TJ-8117) and (-)epicatechin-3-O-gallate on the proliferating changes in glomeruli of 5/6 nephrectomized rats]. AB - Onpi-to (TJ-8117) is a herbal medicine composed of five crude drugs (Rhei Rhizoma, Glycyrrhizae Radix, Ginseng Radix, Zingiberis Rhizoma and Aconiti Tuber). Our previous experiments have demonstrated that TJ-8117 suppressed the development of glomerulosclerosis and retarded the deterioration of renal function in 5/6 nephrectomised rats. In the present study, the effects of TJ-8117 and (-)Epicatechin-3-O-gallate (ECG), which is a component of Rhei Rhizoma, on glomerular cell proliferation, extracellular matrix accumulation and glomerular hypertrophy were investigated in 5/6 nephrectomized rats. Male Wistar rats (170 180 g) were subjected to 5/6 nephrectomy, and TJ-8117 (0.32%, 0.64%) or angiotensin-converting enzyme inhibitor, captopril (CAP 0.08%) was administered daily by mixing in normal chow and ECG (2 mg, 8 mg/100 ml) by drinking water from the day after 5/6 nephrectomy. Following 5/6 nephrectomy, glomerular cell poliferation was increased and reached a maximum at 1 week in the untreated control rats, but was suppressed significantly at 1 and 2 weeks after treatment with TJ-8117 and at only 1 week after treatment with CAP. Extracellular matrix accumulation was detected after 1 week and increased gradually until 4 weeks in the control rats, whereas it was significantly inhibited in both the TJ-8117- and CAP-treated rats. In addition, immunohistochemistry revealed that TJ-8117 significantly inhibited the increase of fibronectin, and tended to reduce type I and type IV collagen at 4 weeks. Furthermore, TJ-8117 suppressed glomerular hypertrophy at 4 weeks. Systolic blood pressure (SBP) and urinary protein excretion (UP) were higher in the control rats than sham-operated rats. TJ-8117 prevented this increase of SBP and UP at 1 week. ECG also suppressed glomerular cell proliferation and the increase of SBP and UP at 1 week after 5/6 nephrectomy. These findings suggest that ECG was one of active components of TJ 8117. These results suggest that TJ-8117 suppressed proliferating changes in glomeruli at an early stage in 5/6 nephrectomized rats, and inhibited the development of glomerulosclerosis. PMID- 9396237 TI - [Ouabain-containing Euro-Collins solution prevents ATN following renal cold storage]. AB - In view of the hypothesis that suppression of energy demand may prevent ischemic cell damage, it seemed possible that suppression of ATP utilization during ischemia might ameliorate the severity of renal failure following kidney preservation. To test this possibility, a short-term in situ kidney preservation model was prepared in dogs. Euro-Collins solution containing 10(-5) M ouabain (O EC) was used as the preservation solution. The kidney was preserved with cold O EC for two hours and reperfused with auto blood. As the control, the kidney was treated with Euro-Collins solution (EC) alone. Three hours after reperfusion, recovery of creatinine clearance was 47.4 +/- 8.0% in the control and 71.6 +/- 14.0% in the O-EC group (p < 0.02). The increase in urinary excretion of N-acetyl beta-D-glucosaminidase was significantly lower in the O-EC group. It was 21.3 +/- 4.5 nU/gr renal weight for three hours after reperfusion in the control group and 7.2 +/- 1.5 nU/gr renal weight in the O-EC group (p < 0.05). Fractional excretion of sodium three hours after reperfusion was 1.42 +/- 0.44% and 5.51 +/- 0.63% in the control and O-EC groups (p < 0.002), respectively. There were no significant differences in renal blood flow, urine volume and urine osmolality between the two groups. These results suggest that ouabain-containing EC was effective in protecting the kidney, especially renal proximal tubular cell, against ischemic damage. PMID- 9396238 TI - [Influence for cultured renal cell growth ability by toxins of Escherichia coli]. AB - There have been many reports indicating that Escherichia coli from pyelonephritis may exhibit several specific phenotypes. The purpose of the present study was to examine the effects of various toxins from Escherichia coli on cultured renal cell growth. alpha-hemolysin suppressed growth of Mardin Darby canine kidney (MDCK) cells in a dose-dependent manner. A low dose of O-antigen suppressed MDCK cell growth, while a high dose of the antigen increased the cell growth slightly. K:1-antigen had no effect on MDCK cell growth. Mesangial cell growth was not affected by alpha-hemolysin. A significant increase in mesangial cell growth was recognized by the O- or K:1-antigen. From these results it can be speculated that alpha-hemolysin from Escherichia coli may play a leading role, and O- or K antigen may play a supporting role in the pathogenesis of pyelonephritis. PMID- 9396239 TI - [Mechanism mediating hypertension induced by chronic inhibition of nitric oxide synthesis]. AB - Although the inhibition of nitric oxide (NO) synthesis is known to induce systemic hypertension, the underlying mechanisms mediating this type of hypertension are incompletely understood. In the present study we investigated the influence of sodium intake on the pressor effect of long-term administration of the NO synthesis inhibitor, NG-nitro-L-arginine methyl ester (L-NAME, 16 mg/dl in drinking fluid for 8 weeks), in conscious Sprague-Dawley rats. Urinary excretion rates of catecholamine during NO synthesis inhibition were also examined. Long-term administration of L-NAME produced a sustained elevation in tail-cuff pressure without altering urine flow, or sodium excretion rate. L-NAME induced hypertension was accompanied by a decreased urinary excretion of the stable NO metabolites, NO2- and NO3-, and was aggravated when rats drank 0.9% saline in place of tap water. Thus, inhibition of NO synthesis resulted in a rightward shift of the pressure natriuresis relationship and a significant decrease in the slope of this relationship. Urinary excretion of epinephrine and norepinephrine, but not that of dopamine, in L-NAME-treated rats significantly increased within the first week of the study when compared with those observed in control rats. A natriuretic index of the sympathetic nervous system, the ratio of dopamine to norepinephrine excretion, was significantly less in L-NAME-treated rats than in control rats. After 8-week treatment with L-NAME, renal morphologic evaluation revealed significant narrowing and obliteration of the arterioles. L arginine (2 g/dl in drinking fluid) completely reversed the elevation of blood pressure as well as the decrease in urinary NO2- and NO3- excretion and the increased urinary excretion of catecholamines associated with L-NAME treatment after 3 weeks of concomitant administration. These results suggest that the inhibition of chronic NO synthesis produces sodium-sensitive hypertension and that changes in sympathetic nerve activity may, at least in part, contribute to the sodium sensitivity in this type of hypertension. PMID- 9396240 TI - [Acquired resistance to acute renal failure in cisplatin-induced renal failure of rats]. AB - Studies were performed to investigate whether prior cisplatin-induced acute renal failure affords resistance to a second challenge with cisplatin in Sprague-Dawley rats. Both the increase in serum creatinine and tubular damage following a challenge with cisplatin (5 mg/kg, i. p.) were significantly less in rats which had received cisplatin (3 mg/kg, i. p.) 14 days prior to the rechallenge compared with the previously untreated animals. Attenuation of nephrotoxicity was more obvious in the histological index than in the increase in serum creatinine, and increase in serum creatinine concentration did not correlate with tubular necrosis. There were fewer tubular cells that expressed proliferating cell nuclear antigen in previously treated kidneys, suggesting that the enhanced regeneration of tubular cells plays a minor role in the decrease of tubular damage in kidneys recovering from prior acute renal failure. These findings suggest that rats recovering from previous acute renal failure are resistant to a second insult with cisplatin and that the attenuation of nephrotoxicity is more prominent in histological damage than in functional disturbance. PMID- 9396241 TI - [HLA-DQ region and TCR gene polymorphism associated with primary IgA nephropathy in Japanese children]. AB - We have investigated associations between HLA-DQ beta and TCRC beta gene polymorphisms and IgA nephropathy in Japanese children. DNA from 33 cases of IgA nephropathy, 29 cases of nephrotic syndrome and 29 cases of healthy volunteers were analyzed by Southern blotting using HLA-DQ beta and TCRC beta probes. A 2 kb (Taq I) polymorphic band that hybridized to the DQ beta probe was predominant in patients of IgA nephropathy (p < 0.05) compared to the controls. The 10 kb (Bgl II) polymorphic band of TCRC beta was also predominantly present (but not significantly) in the DNA of these patients. Proteinuria in IgA nephropathy patients was found to be associated with this 10 kb TCR polymorphism. We conclude that the HLA-DQ beta and TCR genes make major contributions to the genetic pathogenesis of IgA nephropathy in Japanese children. PMID- 9396242 TI - [Role of tumor necrosis factor-alpha in insulin sensitivity and effect of low protein diet on the TNF-alpha response in patients with diabetic renal failure]. AB - In patients with diabetic renal failure, attention must be paid to the prevention of atherosclerosis as well as the preservation of renal function. Insulin resistance is one of the important risk-factors of atherosclerosis and the involvement of tumor necrosis factor-alpha (TNF-alpha) has been shown in the pathogenesis of insulin resistance in some diseases. A low-protein diet (LPD) is recommended for patients with advanced renal disease, but a large proportion of the total caloric intake is supplied from carbohydrates and fat in LPD. Therefore, we designed a study to determine: (1) the effect of TNF-alpha on insulin sensitivity, and (2) the effect of LPD on the TNF-alpha response and the risk factors of atherosclerosis, such as insulin sensitivity and lipid metabolism, in patients with diabetic renal failure. Insulin sensitivity was measured by an euglycemic hyperinsulinemic clamp technique and serum TNF-alpha level and in vitro release of TNF-alpha from peripheral blood mononuclear cells (PBMCs) was measured in patients with diabetic renal failure. A significant negative correlation was observed between lipopolysaccharide-stimulated TNF-alpha release from PBMCs and insulin sensitivity (r = -0.58, p < 0.05). Secondly, risk factors of atherosclerosis were measured before and two weeks after the introduction of LPD in patients with diabetic renal failure. LPD did not have any significant effect on insulin sensitivity, the production of TNF-alpha by PBMCs, lipid metabolism and glucose metabolism. These results indicate that: (1) TNF alpha derived from PBMCs might affect insulin sensitivity in patients with diabetic renal failure, and (2) LPD does not have any significant effect on the risk factors of atherosclerosis. PMID- 9396243 TI - [Implications of obesity for target organ injuries and cardiovascular risk factors in hypertensive subjects]. AB - The effects of obesity on target organ injuries and cardiovascular risk factors were examined in hypertensive subjects. The subjects were 22 obese (OB-HT) and 54 nonobese (NO-HT) men with never-treated essential hypertension, and 37 obese (OB NT) and 50 nonobese (NO-NT) normotensive men. In these 4 groups with the average age of about 50 years, we evaluated serum lipids, glucose tolerance, and hypertensive organ injuries in the heart, kidney, and optic fundus. Although the fasting blood glucose levels were similar in the 4 groups, the area under the blood glucose curve after 75 g glucose ingestion (NO-NT 15.6, OB-NT 17.5, NO-HT 15.8, OB-HT 17.6 x 10(3) mg/dl.min; p < 0.02) and the fast serum insulin level (NO-NT 7.3, OB-NT 10.1, NO-HT 7.7, OB-HT 12.2 mU/l; p < 0.001) were increased in obese men. In OB-HT, serum HDL-cholesterol was decreased (-11%, p < 0.05) and triglycerides were increased (+ 58%, p < 0.01) comparing with NO-NT. The incidence of electrocardiographic left ventricular hypertrophy was not significantly different among the 4 groups, however, urinary albumin excretion was increased in OB-HT (NO-NT 3.0, OB-NT 3.4, NO-HT 3.6, OB-HT 4.3 mg/g creatinine; p < 0.05) and sclerotic lesions of the retinal arteries were observed even in normotensive OB-NT. These data suggest that obesity unfavorably alters lipid and glucose metabolism, and facilitates organ injuries such as arteriosclerosis and renal dysfunction in hypertensive subjects. PMID- 9396244 TI - [An elderly patient with purpura nephritis that appeared after extracorporeal shock wave lithotripsy]. AB - We report a 63-year-old male patient with purpura nephritis, which appeared 7 days after extracorporeal shock wave lithotripsy (ESWL). He was referred to our clinic because of a petechial rash on both lower extremities, pretibial edema and massive proteinuria. Urinalysis showed proteinuria and hematuria and some hyaline casts. A 24-hour urine sample contained 5.0 g of protein. Renal function on admission was decreased: serum creatinine was 1.5 mg/dl and creatinine clearance, 21 ml/min. Immunoserological tests demonstrated an increase in serum IgA (424.3 mg/dl). A skin biopsy revealed leukocytoclastic vasculitis. A renal biopsy showed endocapillary proliferation in a diffuse, but segmental fashion. However, no crescent formation was seen. Immunofluorescence microscopy disclosed mesangial staining for IgA and C3. Electron microscopy demonstrated severe injury to endothelial and epithelial cells: detachment of endothelial and epithelial cells, foot process effacement and macrophage infiltration. Electron-dense deposits were observed in the subendothelial and paramesangial areas. Because renal function was deteriorating rapidly, methylprednisolone pulse therapy and immunosuppressive treatment were implemented. Treatment was effective and the patient's renal function and proteinuria improved remarkably. The electron microscopic findings in this case of purpura nephritis seemed to be more severe than usual, suggesting that ESWL may aggravate glomerular damage. PMID- 9396245 TI - [A case of systemic lupus erythematosus associated with minimal change nephrotic syndrome]. AB - A case of systemic lupus erythematosus (SLE) associated with minimal change nephrotic syndrome (MCNS) in a 25-year-old female is described. The patient suddenly manifested butterfly rash and proteinuria was first pointed out on March, 1994. On admission, her skin biopsy indicated SLE. Subsequently, she developed nephrotic syndrome. Urinalysis showed heavy proteinuria (4.1 g/day), with no other abnormalities in the urinary sediment. Immunological examination revealed positive antinuclear antibody at a titer of 1:80 with a speckled pattern. Anti-ssDNA and anti-SS-A antibodies were positive, but other antibodies were negative. Serum complement (CH50) was within the normal range (30.5 U/ml). The renal biopsy showed no apparent cellular proliferation or increase of extracellular matrices in glomeruli by light microscopy. Slight deposition of IgG, IgM, C3 and C1q was focally seen in the mesangium and capillary wall by immunofluorescence. Electron microscopic examination revealed small and scattered dense deposits in the mesangium, subepithelium and subendothelium, associated with diffuse fusion of the foot processes of epithelial cells along the glomerular basement membrane. According to the WHO classification, the histological features were compatible with those of lupus nephritis (LN), class Ib. The patient was treated with PREDNISOLONE, Mizorbine and Dilazep, resulting in the disappearance of proteinuria and a normal serum level of total protein. The association of LN and MCNS is very rare. We also investigated the relationship between the intensity of proteinuria and histological types of 53 cases with LN examined in our laboratory. The cases with heavy proteinuria were mostly classified as WHO-Class IV and Class V. We report here a case of LN associated with MCNS and also review the literatures. PMID- 9396246 TI - [A case of rapidly progressive IgA nephropathy with transient hypocomplementemia at onset]. AB - We present a case of IgA nephropathy (IgAGN) which developed rapidly progressive glomerulonephritis and showed marked clinical improvement with treatment. The patient was a 7-year-old boy who initially presented with acute nephritic syndrome with hypocomplementemia. Although the renal function improved with normalization of the serum complement level, it deteriorated again progressively. The first renal biopsy revealed cellular crescents in about 70 percent of 43 glomeruli. Immunofluorescent microscopy demonstrated deposits of IgA, C3 and IgG in the mesangium; they were also deposited along the glomerular capillary walls. He was treated with plasma exchange associated with hemodialysis and methylprednisolone pulse therapy, followed by oral administration of prednisolone, cyclophosphamide and warfarin. Renal function recovered to the normal range about two months after the initiation of treatment. The second biopsy demonstrated a marked decrease in histological activity. In this case, transient hypocomplementemia at onset may indicate that acute glomerulonephritis caused exacerbation of clinically silent IgAGN. Aggressive therapy may be effective in patients with rapidly progressive IgAGN if treated at an early stage. PMID- 9396247 TI - [Selective transcatheter embolization for renal artery aneurysms: report of three cases]. AB - Selective transcatheter embolization using an interlocking detachable coil (IDC) or detachable balloon was performed in three patients with renal artery aneurysms. All aneurysms were of the saccular type, located on segmental branches of the renal artery. After the embolization procedures, the levels of LDH, GOT, WBC and the body temperature were transitionally elevated in all patients. Complete occlusion of the aneurysms were achieved in all cases. Good clinical results were achieved in two of these patients without any renal dysfunction. However, local infarction of ipsilateral renal parenchyma occurred in one patient immediately after the embolization and a Tc-99m-DMSA renal scintigraphy study suggested renal dysfunction after infarction, and the level of serum renin activity was slightly elevated. Selective transcatheter embolization may constitute an acceptable therapeutic approach for renal artery aneurysms because beside avoiding surgery, it has a low risk of complications. PMID- 9396248 TI - [Effects of a thromboxane-synthetase inhibitor in patients with chronic persistent coughing and no airwayhyperresponsiveness]. AB - We studied the effects of the thromboxane-synthetase inhibitor ozagrel in 22 patients with chronic persistent coughing who did not have airwayhyperresponsiveness. Treatment with ozagrel (400 mg/day for 2 weeks) reduced coughing in 12 patients. Sputum from the patients in whom ozagrel was effective had a higher percentage of lymphocytes and a lower percentage of neutrophils than did sputum from those in whom ozagrel was not effective. Furthermore, in the former group the capsaicin cough threshold increased but in the latter it did not change consistently. These data indicate that thromboxane A2 may contribute to coughing associated with lymphocytic airway inflammation. PMID- 9396249 TI - [Relationship between exercise-induced hypoxemia and long-term survival in patients with chronic obstructive pulmonary disease]. AB - In patients with chronic obstructive pulmonary disease (COPD), exercise-induced hypoxemia (EIH) is an important limiting factor of exercise performance, but there are very few reports of the prognostic value of EIH. We therefore examined whether EIH is related to long-term outcome in patients with COPD. We gathered data on survival of 77 patients with COPD who had undergone three-minute incremental treadmill exercise tests between 1979 and 1988. A plastic catheter was placed percutaneously into a brachial artery to measure arterial blood gases at each stage of exercise. Expired gas was analyzed continuously during exercise with a Respiromonitor RM-300 (Minato Medical Science). As an index of the severity of EIH we used the delta PaO2/delta VO2 (mmHg/L/min), PaO2-slope. Patients were assigned to two groups according to PaO2-slope, and actuarial survival curves were plotted for the two groups. The survival data were analyzed by generalized Wilcoxon analysis. Survival was significantly better in the group with low PaO2-slopes (< 20 mmHg/L/min) than in the group with high PaO2-slopes (> or = 20 mmHg/ L/min) (p = 0.0031). Also, among the patients in whom PaO2 during exercise was less than 60 mmHg, there was a significant difference in survival between those who received home oxygen therapy and those who did not. We conclude that the degree of EIH can be used to predict survival in patients with COPD, and that the degree of EIH should be considered when prescribing continuous home oxygen therapy for these patients. PMID- 9396250 TI - [Serum-soluble adhesion molecules in patients with idiopathic pulmonary fibrosis and acute respiratory distress syndrome]. AB - We measured the concentrations of soluble adhesion molecules in serum obtained from healthy volunteers and from patients with idiopathic pulmonary fibrosis (IPF) and the acute respiratory distress syndrome (ARDS). The concentrations of soluble intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), E-selectin, and P-selectin were significantly higher in serum from the patients than in serum from the healthy volunteers. In patients with IPF, the concentration of soluble ICAM-1 in serum was inversely related to the vital capacity (expressed as a percent of the predicted value). The concentrations of L-selectin in serum from patients with ARDS were significantly lower than those in serum from healthy volunteers and from patients with IPF. These data suggest that serum-soluble adhesion molecules may be useful as markers of disease activity, severity, and prognosis in ARDS and IPF. PMID- 9396251 TI - [Effect of cessation of erythromycin therapy on diffuse panbronchiolitis]. AB - The effect of cessation of erythromycin (EM) therapy against diffuse panbronchiolitis was studied. Nine cases were examined. After cessation of EM therapy, the manifestations of disease were stable in five cases, but worsened in the other four. In the former five, the period from the onset of disease until EM therapy began was relatively short; when EM therapy was stopped the manifestations of disease had almost completely disappeared and chest roentgenography revealed resolution of diffuse, small, nodular opacities without remarkable bronchiectasis. In contrast, in the latter four cases, the clinical manifestations of disease did not disappear, and chest-roentgenographic evidence of bronchiectasis was common before the cessation of EM therapy. In conclusion, EM therapy for diffuse panbronchiolitis may be stopped if the clinical manifestations of disease (especially purulent sputum) disappear, if diffuse, nodular opacities resolve almost completely and if there is no evidence of bronchiectasis. PMID- 9396252 TI - [Age-related decline in forced expiratory volume in one second and forced vital capacity based on a longitudinal observation of Japanese males]. AB - Forced expiratory volume in one second (FEV1) and forced vital capacity (FVC) of 243 healthy male Japanese workers were measured on as annual basis over seven years, and their longitudinal decline was compared with the age coefficient of cross-sectional prediction equations reported previously for Japanese adults. In this study, a man, assumed to be 1.65 meter tall, was expected to have a respective 22 ml and 11 ml decline annually in FEV1 and FVC. Age-related differences of those indices obtained from cross-sectional prediction equations, however, ranged from 22 ml to 31 ml a year in FEV1 and from 16 ml to 25 ml in FVC for men of the same height. Furthermore, in those equations, age was simply employed as a first-order explanatory variable for ages ranging from the later teens to over sixties, although age-related acceleration of FVC decline is suggested in this study. These results indicate that evaluations of measurements relating to time-series pulmonary functions on cross-sectional prediction equations might be biased. This is probably due to the influence of age-cohort. It seems necessary to build up the reference standards for longitudinal pulmonary function change for an appropriate evaluation of time-series data of FVC and EFV1. PMID- 9396253 TI - [Outcomes of noninvasive positive-pressure ventilation in patients with acute hypercapnia complicating chronic respiratory failure]. AB - We used noninvasive positive-pressure ventilation to treat hypercapnea due to acute exacerbations of chronic respiratory failure (21 episodes in 19 patients; COPD, 4; pulmonary tuberculosis sequelae, 4; silicosis, 3; silicotuberculosis, 3; bronchiectasis, 3; others, 2). All patients had acute onsets of severe hypercapnea (PaCO2 > 45 Torr), acute decreases in pH (< 7.35), and tachypnea, paradoxical breathing or both. During the first 2 to 4 hours of bi-level positive airway pressure, PaCO2 decreased from 72 to 61 Torr (p < 0.0005), pH increased from 7.26 to 7.31 (p < 0.001), and respiratory rate decreased from 30 to 25 breaths/min (p < 0.005). In three cases leakage of air through the mouth prevented improvement in the patients' conditions, but in two of those a face mask was then used successfully. In 17 of the 21 episodes (81%) gas exchange improved and intubation was not necessary. In those 17, the mean duration of noninvasive positive-pressure ventilation was 6.3 days. We conclude that noninvasive positive-pressure ventilation can improve gas exchange in patients with acute hypercapnea complicating chronic respiratory failure. PMID- 9396254 TI - [Inflammatory bronchial polyps with a foreign body-like substance]. AB - A 68-year-old male complaining of hemosputum was admitted to our hospital. Fiberoptic bronchoscopic examinations revealed bronchial polyps and a flat foreign body-like substance in the left main bronchus. A closer inspection of a biopsied bronchial inflammatory polyp showed inflammatory edema with hypervascularization in the submucosal space and inflammatory cell infiltration. Following complete removal of this foreign body-like substance, the polyps disappeared within six weeks. It is therefore feasible to assume that the polyps appeared as a reaction to this extrinsic substance. The origin of the foreign matter is not obvious because the patient had no history of aspiration and because the histological examination did not confirm that it was a foreign body. The substance could have been formed out of the organization and calcification of some secretes in the bronchus. PMID- 9396255 TI - [Fulminant tracheobronchitis caused by methicillin-resistant Staphylococcus aureus (MRSA)]. AB - A 56-year-old man was admitted to our hospital because of hemoptysis, and dyspnea. Breathing sounds decreased and rhonchi was audible in the right lung. Chest X-ray and Chest CT showed infiltrative shadows in the bilateral lower lobes. He was intubated and bronchofiberscopy revealed mucosal necrosis and hemorrhage of the lower trachea and the bilateral bronchi. MRSA was isolated from sputum and bronchial lavage fluids. He was diagnosed as NTB caused by MRSA. On the 7th day in hospital, he suffocated by the necrotic tissue and autopsy was performed. NTB is a very uncommon disease in adults, especially ones who are not under mechanical ventilation. PMID- 9396256 TI - [Two cases of acetaminophen-induced pneumonitis]. AB - This paper deals with two patients with acetaminophen-induced pneumonitis. A 64 year-old woman suffered from mastitis while being treated by corticosteroid therapy for phemphigoid. She was administered antibiotics and acetaminophen. However, her fever continued and she subsequently developed dyspnea and interstitial pneumonia. The other patient, a 70-year-old woman, was treated with corticosteroid for lower motor neuron disease. Anti-GM1-IgM antibodies were positive in her serum. She developed wet cough and mild fever. During treatment with antibiotics and acetaminophen, her illness was complicated by dyspnea and interstital pneumonia. As a result of histological findings of transbronchial lung biopsy specimens showing interstitial infiltration of mononuclear cells, as well as clinical courses in which cessation of acetaminophen directly lead to the improvement of interstitial pneumonia, both patients were diagnosed to have acetaminophen-induced pneumonitis. The peumonitis responded well to steroid therapy. In vitro culture of peripheral lymphocytes showed stimulated proliferation by acetaminophen in both patients. These findings suggest that allergic mechanism was involved in the pathogenesis of the pneumonitis. Underlying immunological disorders may have enhanced the occurrence. Although acetaminophen is one of the most popular drugs because of a very low incidence of side effects, this drug should be applied carefully, especially with patients who have such immunological disorders. PMID- 9396257 TI - [A case of Paragonimus Miyazaki with pleuritis and meningoencephalitis]. AB - A 35-year-old man was admitted to our hospital with fever and headache. Chest X ray revealed right pleural effusion. Lab tests revealed increase of eosinophils in his serum and pleural effusion. After admission he complained of doplopia and neck stiffness. Lumber puncture revealed eosinophilia in the cerbrospinal fluid. Brain CT and MRI showed characteristic images of meningoencephalitis. The patient had eaten raw Potamon dehaani and the case was diagnosed as paragonimus miyazaki after administration of intradermal reaction and Ouchterony's double diffusion test. The patient was successfully treated with praziqantel. It revealed that the pleural effusion and brain edema disappeared chest X-ray and brain MRI. This case can be considered as a characteristic example of Paragonimus Miyazaki with pleuritis and meningoencephalitis. PMID- 9396258 TI - [A case of dyskeratosis congenita with acute interstitial pneumonia]. AB - This is a rare case of Dyskeratosis Congenita (DC) with acute interstitial pneumonia. A 51-year-old man with DC was admitted to our hospital because of cough, sputum and fever. Chest X-ray film showed ground glass opacities in all lung fields for a while steroid's therapy proved effective, but about seven months later the patient's condition became serious. Methylprednisolone, cyclophosphamide and mechanical ventilation therapy were not effective. He died and an autopsy was performed. The lung specimen showed Organizing Diffuse Alveolar Damage, and some parts pointed to bacterial infection. But Pneumocystic carinii pneumonia and Fungal infections were not found. It is therefore necessary to conduct intensive examinations of lung involvement of patients with Dyskeratosis Congenita. PMID- 9396259 TI - [A case of chronic necrotizing pulmonary aspergillosis successfully treated with combination therapy of antifungal drugs and ulinastatin]. AB - A 64-year-old woman with a history of old tuberculosis, had a fungus ball shadow with meniscus sign in the upper right lung field on a chest X-ray film in 1991. Based on the chest X-ray findings, pulmonary aspergilloma was suspected. Because the size of the intracavitary fungus ball increased, the patient was treated with itraconazole over one year in 1995, but there was no improvement. One month later, she was admitted because of fever, hemoptysis and productive cough, and chest X-ray showed an enlargement of intracavitary mass and infiltrative shadow in the right lung. Chronic necrotizing aspergillosis was diagnosed on the basis of her clinical and radiographic features, and positive serological test. Although itraconazol and amphotericin B were given, cavity and intracavitary fungus ball shadow kept growing. Combination therapy of antifungal drugs and ulinastatin markedly improved symptoms and resulted in complete disappearance of the fungus ball on chest CT scan. PMID- 9396260 TI - [A case of malignant mesothelioma of the pleura and the peritoneum detected by sudden onset of back pain and pleural effusion and ascites]. AB - A case of malignant mesothelioma of the pleura and the peritoneum is reported. In April 1996, a 40-year-old men noticed sudden onset of back pain. Radiographic examinations and MRI revealed pleural effusions, ascites, ringed enhanced tumorous lesions in the right posterior diaphragm along the abdominal aorta, and marked thickening of the right diaphragm with moderate signal intensity. On thoracoscopic surgery, there were white small nodules on the intercostal parietal pleura. Tumor cells of a tubulopapillary pattern had large rounded nuclei and eosinophilic cytoplasms in a partially glandlike arrangement. Cytoplasms of tumor cells stained for alcian blue disappeared after hyaluronidase digestion. Immunohistochemical examinations showed positive staining for keratin but negative for CEA. Electron micrographs showed numerous long thin microvilli, desmosomes and intermediate tonofilaments. From these findings, malignant mesothelioma was diagnosed. The malignant mesothelioma cells of the pleura in this case were considered to disseminate the peritoneum directly through the diaphragm or its lymphatic canals. MRI and thoracoscopic surgery were useful for the demonstration of the pleural disseminations and abdominal invasions. PMID- 9396261 TI - [A case of thymic cyst differentiated from cardiac cyst with immunostaining]. AB - This is a case of a 70-year-old woman with thymic cyst which was found by chance on admission for treatment of acute bronchial asthma and diagnosed by pathological examination. After remission of the asthma she was asymptomatic and no pathological physical findings were found except of cervical thyroid tumor of adenomatous goiter. Chest X-ray films revealed a large mediastinal mass with a sharp margin at the right cardiophrenic angle. Chest CT and MRI revealed that it occupied a space from upper anterior mediastinum to right cardiophrenic angle, and that it showed low density area without enhancement, suggesting a mediastinal cyst. A multilocular cyst, 15 cm in diameter, containing serous liquid was excised using thoracotomy. Pathological and immunohistochemical examination of the cyst revealed that its inner surface was lined with benign cylindrical epithelium and that thymic tissues existed in the walls of the cyst, resulting in a thymic cyst. The thymic cyst, of which reports have increased, should be considered in the differential diagnosis of anterior mediastinal cysts. PMID- 9396262 TI - [Long-term survival after multimodal therapy for lung cancer with pleural dissemination]. AB - A 56-year-old man with peripheral T1N0M0 lung cancer underwent thoracotomy. Pleural dissemination was recognized intraoperatively. We performed wedge resection of the lung including the primary tumor, and applied hypotonic cisplatin treatment. We followed this up postoperatively with chemotherapy combined with cisplatin and irinotecan. One year later, a recurrent tumor occurred at the resected end of the lung, so we conducted stereotactic radiosurgery on the recurrent lung lesion, resulting in complete eradication. One year later, no relapse of cancer was observed. We conclude that implementing hypotonic cisplatin treatment in pleural dissemination of lung cancer followed by stereotactic radiosurgery on any recurrent tumor in the peripheral lung was effective for the present case. PMID- 9396263 TI - [A case of tuberculous abscess in the chest wall close to the thickening pleural lesion following tuberculous pleuritis]. AB - A 33-year-old woman with a history of right tuberculous pleuritis was successfully treated in December 1992 by administration of anti-tuberculous drugs, she demonstrated residual localized pleural thickening on chest computed tomography (CT) and gradually developed a subcutaneous mass in the right chest which became apparent in March 1993. In September, chest CT revealed a periocostal abscess in the right anterior chest wall close to the localized pleural thickening. The patient was diagnosed with tuberculous abscess in the right chest wall on confirmation of acid-fast bacilli in a needle aspiration material of the abscess, and was referred to our hospital. Anti-tuberculous chemotherapy was continued but the chest abscess grew, so on January 28, 1994 she underwent a resection of the abscess, the third costal cartilage and bone, and the parietal pleural lesion connected to the abscess. Histopathological examination showed that the abscess and parietal pleural lesion were compatible with tuberculosis, i.e. both lesions consisted of caseous necrosis and epitheloid cell granuloma, but acid-fast bacilli were not demonstrated in both lesions. After one year of postoperative anti-tuberculous chemotherapy, she was followed without any therapy for 3 years and there has been no recurrence to date. When a localized thickening pleural lesion remains after tuberculous pleuritis, complication of tuberculous abscess in the chest wall should be considered. PMID- 9396264 TI - [A case of primary mediastinal choriocarcinoma]. AB - A 27-year-old man with primary mediastinal choriocarcinoma was reported. He was admitted with complaint of right chest pain and dyspnea. Chest X-ray film and computed tomography of the chest revealed a bulky mass at anterior mediastinum and right pleural effusion. Physical examination revealed bilateral gynecomastia, and the serum beta-HCG level was cap at 500 ng/ml. The specimens obtained by percutaneous needle biopsy of the mediastinal mass showed cytotrophoblasts and syncytiotrophoblasts. He received 4 cycles of anti-cancer chemotherapy, and underwent resection for a residual mass, in which viable cancer cells remained in histological examination. In spite of additional chemotherapy, multiple lung metastasis developed rapidly. High dose chemotherapy, with carboplatine (200 mg/m2 x 4 days), etoposide (250 mg/m2 x 4 days) and cyclophosphamide (50 mg/kg x 2 days) was performed in combination with peripheral blood stem cell autotransplantation. However, brain metastasis set in and he died of respiratory failure 9 months after the onset of symptoms. PMID- 9396265 TI - [Resection of multiple thymoma: a case report]. AB - A 74-year-old male, who had been treated for hypertension at the out-patient clinic, was admitted to our hospital because of an abnormal shadow on a chest radiograph. After diagnosis of thymoma by needle biopsy surgery was carried out on July 6, 1995, when an extended thymectomy along with removal of the entire tumor was done. During the surgery it was noticed that there was not one, but two independent tumors located in the anterior mediastinum, the upper left portion was growing from the level of the crania to the left brachiocephalic vein, whereas the lower right portion was growing towards the right thoracic cavity in front of the pericardium. Both tumors were encapsulated firmly, and connected to each other by scantly loose connective tissues. There was no continuity or sarring between the two tumors. Histologically both were diagnosed to be lymphocytic thymoma. We believe that this case is a good example of multiple thymomas and provides evidence of the potential multicentricity of thymoma. It is possible that extended thymectomy may be seeded for a complete resection of thymomas. PMID- 9396266 TI - [Pulmonary sequestration associated with high levels of tumor markers in serum]. AB - A 20-year-old woman was admitted to our hospital because of an abnormal shadow on a chest X-ray film. Laboratory tests done on admission showed high levels of tumor markers in serum. A computed-tomographic scan of the chest showed a multilocular cystic mass in the S10 of the right lung. Angiography revealed an abnormal artery that branched from the abdominal aorta, and therefore pulmonary sequestration was diagnosed. A right lower lobectomy was done. Analysis of fluid from the cyst revealed very high levels of CA19-9, CEA, and SLX. In an immunohistochemical study, epithelial cells of the cyst's walls were stained for CA19-9, CEA, and SLX. After the operation the levels of these tumor markers in serum were almost normal. PMID- 9396268 TI - [Sexual disorder in men]. PMID- 9396267 TI - [Abnormal female reproductive function]. PMID- 9396269 TI - [The genes in the molecular cascade of the mammalian sexual development]. AB - The sex of an individual is established as a consequence of molecular events that occur in the complex genetic cascade. During sexual development, morphological changes are closely associated with the action of genes expression of which is tightly regulated according to the genetic program. Recently, a number of genes that are involved in this process have been isolated. WT-1 and SF-1 are required for the early formation of the gonadal primordium. SRY is the switch to start the male sexual differentiation by triggering the Sertoli cell lineage. An autosomal gene SOX9, which can cause the sex reversal, might also be involved in the male sex determination. SF-1 is a key factor for the male sexual differentiation, regulating the expression of MIS and the synthesis of testosterone in the fetal testis. DAX1 might be responsible for the female sexual development. The mutation or abnormal expression of these regulatory genes and their downstream genes can cause a variety types of the sex reversal and/or malformation of the gonad and the internal and external genitalia. PMID- 9396270 TI - [Organization and regulation of spermatogenesis]. AB - We briefly overview the process as well as hormonal and paracrine regulation of spermatogenesis. Initiation of sperm production occurs under stimulation by FSH and LH. Spermatogenesis can be maintained by testosterone alone at least qualitatively. Stimulatory function of gonadotropic hormones can be modified or modulated on the basis of the local environment present in the testis. Sertoli cells create the blood-testis barrier modulating the entry of substances into the seminiferous tubules. Sertoli cells play the main role in paracrine regulation of spermatogenesis by providing factors required for the development of germ cells. Substances regulating spermatogenesis as paracrine factors in the testis are summarized. PMID- 9396271 TI - [Folliculogenesis and regulating factors]. AB - Folliculogenesis and steroid production are the bases of fertility in the female mammal. Follicle stimulating hormones (FSH) plays an essential role in this process. It is clear that a variety of ovarian autocrine and paracrine factors can regulate this FSH receptor signal. In the past decade, it is established that growth factors, particularly insulin like growth factor and insulin like growth factor binding protein can modulate FSH receptor signal directly or indirectly. In addition to the folliculogenesis, it is possible that growth factors play a key role in follicle selection and atresia. With increased understanding of the intraovarian regulators in folliculogenesis, it becomes clear that several other autocrine paracrine factors may exist in intraovarian regulation of folliculogenesis, including steroid hormones, cytokines, inhibin, activin. PMID- 9396272 TI - [Mechanism on onset of secondary sexual characteristics in the female]. AB - Adrenal androgens begin to increase approximately 2 years before the increase in gonadotropin secretion. This contributes to the growth of pubic hair. The development of the breast is primarily under the control of estrogens by the ovary. Reactivation of hypothalamic LHRH pulse generator is the most important in the onset of puberty. This reactivation initially occurs during sleep, followed by sleep-associated LH release and the increase of estradiol during daytime hours. Thereafter, the secretion of gonadotropin and estradiol increases through puberty and the positive action of estradiol on gonadotropin release is established in the middle to late puberty. Recently, growth hormone has been suggested to excert direct ovarian actions and may be involved in onset of secondary sexual characteristics. PMID- 9396273 TI - [Endocrinological mechanism of puberty in men]. AB - Puberty is a developmental stage during which endocrinological, physical and psycho-social changes proceed. During prepubertal period, hypothalamic-pituitary gonadal axis is maintained in an inactive state by negative feedback regulation or inhibitory function of central nervous system. The onset of puberty and sexual maturation are gained by the alternation of these inhibitory system. In recent years, highly sensitive techniques for evaluating the hormones of hypothalamic pituitary-gonadal system have been developed. And many explorations of the hormonal mechanisms of puberty have been achieved. In this chapter, hormonal changes in puberty is reviewed, and mechanism of puberty in men is discussed. PMID- 9396274 TI - [The regulation mechanism of the female menstrual cycle]. AB - The hormonal patterns during menstrual cycle, which consist of cyclic alterations in gonadotropins, estradiol, and progesterone, are controlled by hypothalamic pituitary-ovarian feedback mechanism. GnRH produced in hypothalamus acts on the pituitary cells to secrete FSH and LH, which stimulate the follicular development. The developed follicles secrete estradiol, progesterone, inhibin, activin, and follistatin. Estradiol and progesterone, at different concentrations and/or ratios, either positively or negatively control the feedback of hypothalamic-pituitary axis in regulating the secretion of GnRH, FSH and LH. Inhibin and follistatin selectively suppress, whereas activin enhances the secretion of FSH in the pituitary. Recently, various additional factors produced by the ovary have been identified to contribute to the follicular development by paracrine and/or autocrine regulation as well as to feedback on hypothalamic pituitary unit. PMID- 9396275 TI - [Regulation of male sexual function]. AB - MECHANISM OF ERECTION: Commands transmitted from the brain to the spinal cord during audiovisual sexual stimulation and sexual fantasy cause excitation of the central erection-related nerves of the spinal cord, which transmit the commands to the corpora cavernosa. As a result, the following sequence of phenomena takes place: 1) The cavernosal and helical arterioles dilate and the volume of blood in the lacunar spaces increases. 2) The cavernosal smooth muscle relaxes and the vascular lacunae expand. 3) Extension of the lacunar spaces causes compression of the veins below the albuginea corpora cavernosa of the penis. MECHANISM OF EJACULATION: The mechanism of ejaculation can be divided broadly into two stages. In the first stage, seminal emission to the posterior urethra and closure of the internal urethral opening are occurred by stimulation of the hypogastric nerve. The second stage of ejaculation, which is projectile ejaculation of the semen from the posterior urethra, occur as the result of a spinal reflex via the pudendal nerve. For this sequence of events to occur, simultaneous stimulation from the hypogastric nerve is required. PMID- 9396276 TI - [Regulation of gonadotropin secretion]. AB - Physiological gonadotropin levels are modulated by complex relationships between sex steroids and the hypothalamic gonadotropin-releasing hormone (GnRH) pulses. The frequency and amplitude of GnRH secretion provide signals for the differential regulation of LH and FSH secretion. At higher GnRH pulse frequencies, LH secretion increases more than FSH secretion, whereas, at lower frequencies, FSH secretion is favored. Gonadotropin secretion and subunit gene expression are regulated by sex steroids acting either directly at the pituitary level or indirectly by alteration of GnRH pulses from the hypothalamus. And sex steroids have positive or negative actions, depending on the model system and physiological state. Three gonadally derived peptides, inhibin, activin, and follistatin have been isolated and shown to have specific effects on FSH gene expression. Although gonadotropin regulation and reproductive function have not been fully determined, current molecular technologies will probably identify additional targets of sex steroids and GnRH action and allow greater insight. PMID- 9396277 TI - [Androgen--biosynthesis, receptor and action]. AB - Androgens play a key role in promoting normal sex differentiation and development, pubertal masculinization, initiation of spermatogenesis, and maintenance of male sexual function. In addition to these classical function, androgens are significantly associated with cell differentiation, proliferation as well as carcinogenesis. The function of androgens can be mediated by the androgen receptor (AR), which transduces the steroid signal within cells. AR belongs to the superfamily of nuclear receptors that employ complex genetic mechanisms to control the development and physiological functions of target tissues. AR activates or represses gene transcription through association with specific DNA elements and/or proteins. Moreover, molecular investigations of AR gene structure have provided new insights towards defining a genetic basis for the relationship between the diseased conditions and androgen action. Recent data on androgen biosynthesis, action of androgens and molecular genetic analysis of gene structure have led to a new understanding of the pathology in affected men. This review summarizes briefly our current view of androgen action with a special emphasis on the alterations of AR. PMID- 9396278 TI - [Estrogen, progesterone--biosynthesis, receptor and action]. AB - Ovarian sex steroid hormones, estrogen and progesterone, have crucial effects on female reproductive functions such as the development and maturation of reproductive organs and the appearance of normal menstrual cycles. The precursor of steroid hormones is a cholesterol that is synthesized de novo from acetate or that is taken in as LDL-cholesterol via LDL-receptor. Members of cytochrome P450 superfamily and hydroxysteroid dehydrogenase (HSD) involved in the biosynthesis of the steroid hormones. In the corpus luteum, P450scc and 3 beta-HSD catalyze the pathway from cholesterol to progesterone and in the granulosa cells, P450arom (aromatase) and two HSDs catalyze the biosynthesis of estrogen from C19 steroid synthesized in the theca cells having P450(17) alpha. In the target cell, steroid binds to and activates its receptor located primarily in the nucleus. Binding of ligand results in dimerization of the receptor due to its conformational change. The steroid-receptor complex binds to specific hormone responsive element of the target gene at the site of DNA binding region and then activates transcription of the gene. Main action of estrogen is to stimulate the proliferation and development of cells due to enhancement of the synthesis of specific DNAs and proteins in the female reproductive organs. Progesterone regulates the action of estrogen through the decrease of estrogen receptor and down regulation of its own receptor. PMID- 9396279 TI - [Physiology and action of prolactin]. AB - Prolactin (PRL) is a peptide hormone secreted by the lactotroph of the anterior pituitary. Secretion of PRL is regulated negatively by hypothalamic hormone. Dopamine is the major physiologic prolactin inhibiting factor (PIF). PRFs as TRH, VIP and oxytosin have prolactin releasing activity. Prolactin secretion is induced by sleep, stress, sexual intercourse and suckling stimulus. Serum level of PRL is increased by estrogen. Therefore, PRL level is higher in women, especially in the late follicular phase and during pregnancy. Lactation is the major biologic function of PRL in humans and essential to reproduction. Other function of PRL, such as regulation of ovarian function, osmoregulation and immunoregulation is not fully established. Many action of PRL remained to be clarified. PMID- 9396280 TI - [Abnormal female reproductive function]. AB - Sex differentiation may be seen as a sequence of gonadal determination, gonadal differentiation, genital differentiation. Malfunction of this causes abnormal female reproductive function. SRY (sex-determining region Y) has been show to induce testis which is needed to bring about development of the other male sex organs. Absence or point or frame shift mutation in the SRY causes XY females. Secondary sex determination depends on testosterone produced by Leydig cells. Testicular feminization syndrome typically lack the normal androgen receptor protein and therefore, they are distinctly female phenotype. Rokitansky syndrome is a type of aplasia vaginae and is a malformation of the Mullerian duct. They all present primary amenorrhea. Secondary amenorrhea is a common type of abnormal female reproductive function and differential diagnosis depends on hormonal analysis. One of the topics includes polycystic ovarian syndrome which is recently treated by laparoscopic surgery. PMID- 9396281 TI - [Definition and classification of sexual disorder--in men]. AB - Sexual disorders include the disorder of sexual differntiation and male infertility and sexual dysfunction. Sex determination is concerned with the control of the development of the primary and or gonadal sex, sex differentiation encompasses the events subsequent to gonadal organogenesis. Classification of anomalous sexual development are recognized as disorders of gonadal differentiation, female pseudhermaphroditism, male pseudhermaphroditism and unclassified form. Upon completion of a history and physical examination and evaluation of semen parameters, the male infertility can usually classified into one of nine categories: azoospermia, ejaculatory dysfunction, varicocele, gonadotoxins, antisperm antibodies, infections, endocrinopathy, idiopathic infertility, or miscellaneous. Erectile dysfunction (ED) is classified into two categories: functional ED and organic ED. Vascular, neurological and endocrinological disorders are recognized as organic causes of ED. PMID- 9396282 TI - [Virilization syndrome and hirsutism]. AB - Severe virilization syndrome is seen in testosterone-producing ovarian or adrenal tumor while symptom in late-onset congenital adrenal hyperplasia (CAH) are usually mild hirsutism and amenorrhea. We determined the serum levels of 17 alpha hydroxy pregnenolone (17P5) and 17 alpha-hydroxyprogesterone (17P4) 60 min after the intravenous injection of 250 micrograms of ACTH in 10 normal Japanese females with age ranging from 18 to 29 years old. In this test, 1 mg of dexamethasone had been administered orally at 11 p.m. on the previous day to suppress the effect of endogenous ACTH. The ratio of 17P5 to 17P4 was 10.0 +/- 3.6 (mean +/- SD). Five females with hirsutism of no ovarian origin showed normal responses to this rapid ACTH test, suggesting that the incidence of the late-onset CAH is not so high as reported previously. PMID- 9396283 TI - [Feminizing syndrome and gynecomastia in males]. AB - Gynecomastia is one of the common symptoms of feminizing syndrome in males. Causes of feminizing syndrome are considered the increase of serum level of estrogen, prolactin, LH and hCG and testosterone deficiency. The condition of gynecomastia was commonly silent. Gynecomastia is classified three groups those are physiological, pathological and ideopathic gynecomastia. Physiological gynecomastia was reported to break out healthy males. Ideopathic gynecomastia is the main cause in medicine. Characterization of pathological gynecomastia are known side effect of drugs, testosterone deficiency and increased estrogen production. Gynecomastia is making satisfactory progress by treatment of the causes, but long term of the history merely have need resection. PMID- 9396284 TI - [Sexual precocity]. AB - Sexual precocity results from both GnRH-dependent and GnRH-independent mechanisms. The GnRH-dependent forms of precocious puberty can be treated effectively with long-acting agonist analogues of GnRH. However, for some children who have a poor growth rate during the analogue therapy, an additional growth hormone therapy should be considered to get them near to their normal final height. The analogue treatment could be continued until a bone age of 13 or more. The GnRH-independent forms of precocious puberty have unique mechanisms and unknown pathophysiology. For instance, even though it is known that testotoxicosis and McCune Albright syndrome are caused by the molecular mechanism disorders, further elucidation of their etiologic basis of sexual precocity is needed. Thus being, in treating the GnRH-independent forms they should be assessed for each particular disorder. PMID- 9396285 TI - [Gonadal dysfunction]. AB - Function of hypothalamic-pituitary-ovarian axis is an essential factor for the maintenance of regular cycles in mature women. The disturbance of function of those organs causes gonadal dysfunction such as anovulation, amenorrhea and menstrual disorders. Therefore, the correct diagnosis for the assessment of CNS and ovarian function is clinically important to treat the patients those who have an menstrual disorders. In this review, the mechanism of normal gonadal cycles and the diagnostic method and the treatment of gonadal dysfunction are described. PMID- 9396286 TI - [Male sexual insufficiency]. AB - There are patients who complaint the loss of sexual desire, loss of erection, absence of emission, absence of orgasm, and premature ejaculation. When the primary disturbance is in the producing or releasing process of testosterone in the testis, it increases the gonadotropic stimulation and results in the hypergonadotropic hypogonadism. Hypogonadotropic hypogonadism, in contrast, suggests that the disturbance in the hypophysis or hypothalamus. Patients who have the primary disturbance in the hypothalamus sometimes do not respond to LHRH, but repeated pulsatile LHRH stimulation induces the response. LH testosterone is known to be the main axis of sexual function. However, growth hormone (GH) also has an important role. GH is necessary for adults, not only for children. Its deficiency arises easy fatiguability, loss of sexual desire, loss of erection, and oligo- or azoospermia. Erectile dysfunction is frequently caused by vascular, neurological and psychosomatic disorders. A variety of drugs are known to cause male sexual insufficiency. Many environmental factors and stress may affect the release of LH and GH, and therefore disturb the male sexual function. PMID- 9396287 TI - [Galactorrhea]. AB - Galactorrhea syndromes are mainly caused by hyperprolactinemia, which has been defined by the basal prolactin level more than 15 ng/ml. However, normoprolactinemia can not be proved only by the basal prolactin level less than 15 ng/ml, which required the assessment of prolactin secreting capacity. Occulted hyperprolactinemia has be well known as the same syndrome as hyperprolactinemia, which shows basal prolactin level less than 15 ng/ml and the exceed response of prolactin to prolactin secreting stimulation like as thyroid releasing hormone. Women with occulted hyperprolactinemiais show temporary and intermitted hyperprolactincmia responding to a lot of atimulous states like as stress, sleep or elevated E2 level, which resulted in galactrrhea, menstrual disturbances or infertility. The elevated prolactin not only suppress the pituitary ganadotropin secretion, but also disturb follicular development and luteal function in the ovary. Dopamine agonists, bromocriptine and teruguride an usually indicate in the treatment of hyperptolactinemia and have brought the good results. PMID- 9396288 TI - [Male pseudohermaphroditism]. AB - Male pseudohermaphroditism is a condition of sex differentiation disorders in which the gonads are tests, but the genital ducts and/or external genitalia are incompletely masculinized. This syndrome is caused by a failure of the sequential process in embryonal development of the testis. In the presence of functioning testis the Mullerian ducts regress, while the mesonephric ducts and urogenital sinus differentiate into the internal and external male genitalia. Male pseudohermaphroditism is classified to subtypes according to etiological factors: (1) testicular unresponsiveness to hCG and LH; (2) defect in testosterone biosynthesis; (3) end-organ resistance to androgen; (4) defects in the intracellular metabolism of testosterone; (5) aberrations in testicular organogenesis; (6) defects in anti-Mullerian hormone. PMID- 9396289 TI - [Female pseudohermaphroditism]. AB - Patients with female pseudohermaphroditism have female internal genitalia and karyotype (XX) and various degree of external genitalia virilization. External genitalia is musculinized congenitally when female fetus is exposed to excess androgenic environment. Congenital adrenal hyperplasia (CAH), mostly 21 hydroxylase deficiency, is the most common cause. Maternal androgen excess due to maternal ovarian tumor or drug intake also causes female pseudohermaphroditism. Combination of hormonal therapy and surgical correction is required for CAH. When appropriate treatments were given, normal puberty, fertility and child bearing are possible. HLA typing in patient's family is useful for identifying heterozygote and homozygote, because of close linkage of 21-hydroxylase gene and HLA gene. Prenatal diagnosis and genetic diagnosis for female pseudohermaphroditism due to 21-hydroxylase deficiency can be performed, however prenatal treatment is not completely established. PMID- 9396290 TI - [Male sterility]. AB - Recently several studies have suggested a decline in the quality of semen. About half of the infertile causes are in men, the rate is increasing in the infertile couples. There are some therapy for the male sterility; medication, surgery or assisted reproductive technology (ART). Medicinal effects are not expected, and surgical cases are localized for indication. Moreover, since most of male sterility are idiopathic insufficiency of spermatogenesis, a recent tendency in the male sterile therapy is ART such as IVF-ET, ICSI, TESE, etc. PMID- 9396291 TI - [Female sterility]. AB - Recent progress in female sterility of Japanese women is reported in this study. The causes of sterility in the female side, the most popular sterile factor is a tubal factor due to the increase of the endometriosis and pelvic inflammatory disease. The main treatment of the tubal factor is an IVF-ET technique. The ovulatory disturbances occupy the second position of the female sterile factor. The precise endocrinological examinations must be done for the diagnosis of the ovulatory disturbances. In case of the hypergonadotropic hypogonadism, the ovulation induction is almost impossible. Generally, clomiphene citrate is used for moderate hypothalamic anovulations, and bromocriptine or terguride is for prolactin related diseases (hyperprolactinemia, occult hyperprolactinemia, galactorrhea) and endocrinological PCO (LH/FSH > 1) in the beginning. If these treatments are not effective, hMG-hCG therapy will be performed. During hMG-hCG therapy, we must take care for the occurrence of the ovarian hyperstimulation syndrome. For the treatment toward the cervical factor, the artificial insemination with husband semen will be done, however, if it is not effective, IVF-ET must be performed. For the uterine factor (such as after hysterectomy state, adhesion of uterine cavity, etc.) surrogate mothers are hired in some foreign countries, however, it is prohibited in Japan. If patients have anti phospholipid antibodies, low dose aspirin and/or heparin administration will be done for the treatment of infertility. Recently, the assisted reproductive technology (ART) have made much progress. IVF-ET is performed widely in Japan. Cryopreservation of the fertilized ovum and intracytoplasmic sperm injection (ICSI) are also performed. However, the legal and ethical problems are occurring, and much discussion must be done for the acceptance of such new technologies. PMID- 9396292 TI - [Erectile dysfunction]. PMID- 9396293 TI - [Idiopathic delayed puberty--female]. AB - The developmental changes during puberty occur over a period of 3 to 5 years, usually between ages 9 and 14. The age of onset of puberty is influenced by genetic and environmental factors. Delayed puberty is a rare condition in girls, and a genetic problem or hypothalamic-pituitary-ovarian disorder can be suspected. In addition, anatomic abnormalities of the uterus and ovaries are rare but important factors to consider. The history and physical examination are useful in the diagnostic work-up. Pelvic examination and U.S. should be completed. Also, hormonal analysis should be recommended with diagnostic plan. Although, the possibility of rare diagnoses should always be kept in mind. PMID- 9396294 TI - [Constitutional delay of growth and puberty in male]. AB - Constitutional delay of growth and puberty (CDGP), featuring short stature, delayed puberty and delayed bone age, is the most common condition in pediatric endocrine clinic and becomes an important differential diagnosis in boys with short stature. The conventional management is to assure eventual development of puberty and normal final stature. Treatment with androgens to induce secondary sex characteristics is given only when the boy is under psychosocial stress. Recent reports revealed poor final height outcomes. The results of growth hormone treatment in idiopathic short stature, including CDGP, has not been successful. New findings showed that adults with a history of delay puberty had significant osteopenia and may have a risk of fracture, and that suppression of spinal growth was closely related to pubertal delay, leading to short stature. These two facts suggest the necessity of induction of puberty in normal timing. PMID- 9396295 TI - [Familial male-limited precocious puberty]. AB - Familial male-limited precocious puberty (FMPP) is a gonadotropin-independent disorder that is inherited in an autosomal dominant, male-limited pattern. Affected males generally exhibit signs of puberty by the age of 4 years. Testosterone production and Leydig cell hyperplasia occur autonomously in context of prepubertal levels of luteinizing hormone (LH). The LH receptor is a member of the family of G-protein-coupled receptors. Thus far, eleven constitutively activating mutations of the LH receptor gene, which result in high basal cyclic AMP levels, have been identified in pedigree with FMPP and in sporadic cases. It has been suggested that some of the mutations have effects on the Gq coupling and phospholipase-C activation in addition to their effects on Gs coupling and activation of adenylate cyclase. PMID- 9396296 TI - [Disorders of sex chromosome]. AB - Disorders of sex chromosome, X and Y, consist of abnormality of the number or structure of the sex chromosome. Because sex chromosomes have a variety of genes related to sexual differentiation, disorders of sex chromosome induce a variety of disorders of sexual differentiation. At first, in this title, the process of normal sexual differentiation is shown. Classical disorders of sex chromosome are Klinefelter syndrome, XX male, XYY male, Turner syndrome, XXX female, and XY female. True hermphroiditism, mixed gonadal dysgenesis, and pure gonadal dysgenesis are also included, because most of these disorders have abnormal sex chromosome. Molecular analysis of sex chromosome is clarifying disorders with a minute abnormality of sex chromosome. They include male infertility, premature ovarian failure, and fragile X syndrome. Explanations of the above disorders are given briefly. PMID- 9396297 TI - [Disorders of gonadal function in hypothalamic-pituitary diseases]. AB - Hypothalmic-pituitary-gonadal hormones are regulated by number of factors including GnRH, LH and FSH, and gonadal hormones. They are regulated also by pituitary prolactin, activin and inhibin. Hypogonadism is divided into primary and secondary hypogonadism. Secondary hypogonadism is caused by various lesions involving either the hypothalmus or the pituitary. In this paper, etiology, diagnosis and treatment of the secondary hypogonadism are described. The most frequent causes of hypothamic hypogonadism are hypothalamic tumors, such as germinoma. As the pituitary hypogonadism, pituitary tumors, and Sheehan's syndrome are most common disorders. Diagnosis of the secondary hypogonadism is made based on the diagnostic criteria established by The Research Group of the Japanese Ministry of Health and Welfare. The criteria includes signs and symptoms in addition to laboratory examinations such as measurement of serum gonadotropin, sex steroid and LH-RH test. First choice of the treatment of hyothalmic hypogondism is continuous pulsatile injection of synthetic LH-RH. In the pituitary hypogonadism, hCG, hMG, estrogen and testosterone or the combination of these are applied depending upon the age and other factors of the patients. PMID- 9396298 TI - [Thyroid disease and reproduction dysfunction]. AB - Thyroid disorders have been implicated in a broad spectrum of reproductive disorders ranging from abnormal sexual development to menstrual irregularities and infertility. Hyperthyroidism in the male is thought to cause gynecomastia. In the female hypo and hyperthyroidism results in changes in cycles length and amount of bleeding. The hypothyroidism in the male has less clear-cut effect on the reproductive system. Long-standing, untreated hypothyroidism is associated with galactorrhea. These abnormalities are reversible with adequate thyroid supplementation or collection of hyperthyroidism. Thus during the investigation of hirsurism, menstrual irregularity, infertility, galactorrhea, and gynecomastia, the possibility of thyroid dysfunction must always be considered. PMID- 9396299 TI - [Sexual and gonadal dysfunction in adrenal disorders]. AB - Among various diseases of the adrenals, major disorders that cause sexual and gonadal disturbances are congenital adrenal hyperplasia(CAH) and Cushing's syndrome, and the others include virilizing or feminizing adrenocortical tumors. CAH was reviewed based on the recent advances in molecular genetics. The most striking discovery was steroidogenic acute regulatory protein, mutations of which produce lipoid adrenal hyperplasia that was previously attributed to P-450scc deficiency. Reversible amenorrhea or impotence is found in patients with Cushing's syndrome. Low plasma estrogen and testosterone levels are associated with female and male patients, respectively. Elevated adrenal androgen accounts for mild virilization in female patients with ACTH-dependent subtypes. The sites of action at which hypercortisolemia suppresses the hypothalamic-pituitary gonadal axis were discussed. PMID- 9396300 TI - [Gender issues and eating disorders]. AB - Sexual problems are not specific for eating disorders. The etiology is complex and no one single causal facter has been identified. However, clinical as well as epidemiological studies have shown that eating disorders occur more commonly in females than males. The evidence that eating disorders are more common in females has resulted in the postulation that socio-cultural factors may be important. An important aspect of the socio-cultural position of women which may contribute to eating disorders is the conflict in roles. Clinical experience and research have shown the important role of sexual problems and traumas in the development of anorexia nervosa and bulimia. When compared to anorexics, bulimics reported greater sexual interest and activity. PMID- 9396301 TI - [Sexual dysfunction in diabetes mellitus]. AB - Erectile dysfunction or impotence is a very common complication in diabetic male patients; the prevalence of which may be more than that of retinopathy. The cause of diabetic impotence has been thought to be neuropathy or angiopathy or both of them. The diagnosis of diabetic impotence is based on the exclusion of other causes of impotence including psychological, iatrogenic, endocrinological impotence. The treatment options for diabetic impotence such as vacuum device, intracavernous self-injection or surgical procedures are available and useful at present. In this article, other sexual dysfunction; retrograde ejaculation and female sexual dysfunction in diabetes mellitus are also discussed. PMID- 9396302 TI - [Sexual dysfunction in the male patients on hemodialysis]. AB - Several features of sexual dysfunction, such as infertility, decreased libido and potency, are frequently observed in male patients with uremia, and it usually worsens with time despite of hemodialysis(HD) therapy. Hormonal profile often demonstrates hypergonadotropic hypogonadism, and is suggestive of primary Leydig cell dysfunction. Hypotestosteronemia and hyperprolactinemia may partially participate in the pathogenesis of sexual dysfunction. Uremic toxins, renal anemia, hyperparathyroidism, zinc deficiency, vascular and neurologic abnormalities are also reported to be the causative factors of sexual dysfunction. Correction of anemia with recombinant human erythropoietin sometimes results in the amelioration of sexual potency, probably due to improvement of erectile performance by increased blood viscosity. Psychological derangement should be kept in mind as an another factor of sexual dysfunction. PMID- 9396303 TI - [Endocrine disturbances in liver cirrhosis--focused on sex hormones]. AB - Various endocrine disturbances are often observed in the patients with liver cirrhosis. We focused this paper on the sex hormones. Clinical features of male cirrhotic subjects are feminization(gynecomastia etc) and hypogonadism(testicular atrophy, reduced fertility, loss of libido, impotence etc). Chief abnormalities of sex hormones are a decrease in serum testosterone levels and an increase in serum estrogen levels accompanied by an increase in ratio of estrogen to testosterone in the patients with severe liver cirrhosis associated with the severity of hepatic dysfunction. Hyperestrogenization may be related with feminization of male cirrhotic subjects, whereas hypogonadism is the result of alcohol abuse per se, rather than the indirect consequence of liver cirrhosis. PMID- 9396304 TI - [Obesity and disturbance of sexual function]. AB - Insulin-resistance is a central character of the simple obesity, resulting from decrease of the insulin receptor and dysfunctions of the post receptor systems. It induces hyper-insulinemia. Excess insulin gives rise to the synthesis of androgens by binding to the receptor of the ovary, and suppresses the SHBG and IGFBG in the liver. As a result, in obese women, the sexual function is disturbed by hyperandrogenic milieu. The conversion of androgens to estrone in the lipocytes evokes overproduction of estrone, which stimulates LH release and induces overproduction of androgens. Hyperandrogenemia is a cause of disturbance of the sexual function in obese women. PMID- 9396305 TI - [Polycystic ovary syndrome: PCOS]. AB - Polycystic Ovary Syndrome(PCOS) was originally reported by Stein and Leventhal in 1935, as an syndrome with bilateral polycystic ovaries, menstrual abnormality, hirsutism and obesity etc. After this report the diagnosis of "Polycystic Ovary" was abused for the patient only with polycystic change of ovaries, so that, the concept or definition of PCOS has became unclear and controversial. In this review, a classification of PCOS according to the presence of hyperandrogenemia and/or hirsutism will be shown to reconstruct the concept or definition of PCOS. And usefulness of this classification will be revealed by showing endocrinological and morphological aspect of each class. Furthermore, new therapeutic approaches for PCOS with pure FSH injection in combination with GnRH analog against the onset of OHSS, or drillings of antral follicles with CO2 or KTP laser will be also shown in this review. PMID- 9396306 TI - [Cryptorchidism]. AB - Cryptorchidism represents the most common genital abnormalities in boys. Cryptorchidism means the abnormalities that lead to the empty scrotum and specifically refers to undescended testis. Testicular descent is multifactorial, and abnormalities of morphogenesis and hormonal aspects are considered to be the etiology of non-descent of the testis. Furthermore, hormonal abnormalities associated with cryptorchidism have the potential risks of infertility and malignancy that appear in later life. The modality of the management has undergone major refinements with the introduction of laparoscopy. Laparoscopy has become the most widely used and most useful diagnostic modality in the management of the impalpable undescended testes. PMID- 9396307 TI - [Sexual dysfunction]. AB - According to the definition of the ICD-10 or DSM-III, sexual dysfunction is not caused by organic disorder or disease, and psychogenic. But, the patient who is suffered from sexual dysfunction caused by organic disorder or disease, has many psycho-social problems. So, in this article, all types of sexual dysfunction are subjected. Especially sexual hypofunction is mentioned. And the new type of sexual dysfunctions such as computer sex are also explained. Sex therapy of the Masters and Johnson method and the new sex therapy of Kaplan, H. are explained. PMID- 9396308 TI - [Sexual dysfunction following pelvic surgery]. AB - In male, sexual dysfunction was a common complication that occurred after radical pelvic surgery: radical protectomy, radical cysto-, prostatectomy. Upon the recent pelvic neuroanatomical findings and preservation of these nerves, it is now possible to perform successful cancer operation on the rectum, prostate or bladder with preservation of sexual function in the group of early cancer patients. Depending on the location and severity of these nerve injury, this could result in temporary or permanent erectile and ejaculation dysfunction. In female, the total hysterectomy for cervical cancer sacrifices or injuries the faculty of pregnancy or sexual intercourse. The oophorectomies causes a deficiency of female hormones. But recently the numbers of patients with a small or early stages cancer of uterine or ovary are increasing and we have become to be able to save the functions of these organs in many patients well with minimum local excision or partial resection of them. PMID- 9396309 TI - [A study on sexual dysfunction in female patients with alcoholics]. AB - Few investigations have been made concerning hormonal changes and dyspareunia in fertile aged women with alcoholics experiencing sexual dysfunction. Twenty-seven Japanese woman with alcoholics under 40 years of age excluded with liver cirrhosis were studied to describe alcohol drinking related to sexual dysfunction. Among 21 sexually active women, 20(95.2%) had both symptoms of dyspareunia and vaginal dryness, and only one had neither symptom. Most of patients have lower estradiol levels and 92.0% of patients have the moderately elevated prolactin levels. Eleven of them were having the second grade amenorrhea associated with hyperprolactinemia and hypergonadotropic hypogonadism and 14 were having the first grade amenorrhea. In this study alcoholic abuse women may have deeply related to the hyperprolactinemia, dyspareunia, amenorrhoea, vaginal dryness, ovarian dysfunction and fetal alcohol syndrome. PMID- 9396310 TI - [Alcoholic effect on male sexual function]. AB - Though an adequate volume of ethanol relieves nervousness and enhances sexual desire, acute administration of a great deal of ethanol suppresses central nervous system and causes sensory torpor and penile erectile dysfunction. Long term and excessive intake of ethanol causes central and/or peripheral neuropathy and sexual dysfunction; atrophy of testicles, low serum level of testosterone, impaired spermatogenesis and penile erectile dysfunction. It also invades various organs in digestive tract, cardiovascular system, central and peripheral nervous system and causes functional disorders in these organs. Successful treatment of patients with penile erectile dysfunction should be performed with treatment of these underlying and associated disease. PMID- 9396311 TI - [Drug induced impotence]. AB - Medication induced impotence has been reported in the some papers. However, it is unusual to talk about the sexual history in a general clinical interview and there are few reports to show the possible mechanisms of induced sexual dysfunction in Japan. Recently, the average life span of the Japanese population has gradually increased, so now we have a chance to evaluate the sexual history from older people. Some drugs, such as diuretics, antiarrtythmic and antihypertensive drugs, were reported to induce impotence, however, there was not a detailed study in Japan. We studied the effect of cardiovascular medicine to sexual dysfunction in 1,000 patients. Almost all medication did not always reduce sexual activities, however, sexual activities in old people might decrease because of medications or sickness. It is important for the general physician to ask the patients about general conditions including sexual history while on medications. PMID- 9396312 TI - [Hepatitis C virus as a causative agent of carcinogenesis--recent trends for study]. AB - Hepatitis C virus (HCV) infection causes chronic hepatitis, cirrhosis and hepatocellular carcinoma. However, the mechanism of carcinogenesis by HCV is poorly understood. In this paper, the recent virological and molecular biological studies of HCV published in after 1996 were summarized including the past reported data. Major topics of the study are the biological functions of HCV proteins and the interaction between HCV proteins and cellular proteins. HCV replication and proliferation systems using human cultured cells were also described. The possible role of HCV for development of hepatocellular carcinoma is discussed. PMID- 9396313 TI - [Demonstration on Borna disease virus in patients with chronic fatigue syndrome]. AB - Chronic fatigue syndrome (CFS), a recently named heterogeneous disorder, is an illness of unknown etiology. The association between CFS and several viral infection has been suggested. Here, we centered on the possible link between CFS and Borna disease virus (BDV) infection. BDV is a neurotropic, nonsegmented negative-strand (NNS) RNA virus. Recent epidemiological data have suggested that BDV may be closely associated with depression and schizophrenia in humans. In Japanese patients with CFS, the prevalence of BDV infection was 34% (30/89) and 12% (7/57) by immunoblotting and PCR analysis, respectively. Furthermore, anti BDV antibodies and BDV RNA were detected in a family cluster with CFS. These results suggested that this virus contributes to or initiates CFS, although the single etiologic role of BDV is unlikely. PMID- 9396315 TI - [Diabetes mellitus in the elderly]. PMID- 9396314 TI - [Recent advance in research of Alzheimer's disease]. PMID- 9396316 TI - [Usefulness of dipyridamole thallium scintigraphy in the diagnosis of coronary artery disease in elderly patients]. AB - Patients suspected of having coronary artery disease (CAD) underwent dipyridamole thallium scintigraphy, exercise electrocardiography, and coronary angiography. Of the 500 patients studied, 163 were at least 65 years old, and 337 were less than 65 years old. Both CAD and multivessel CAD were more common in elderly than in younger patients (81% vs 69%, p < 0.01 for CAD; and 43% vs 26% p < 0.01 for multivessel CAD). In patients without myocardial infarction, the specificity of exercise electrocardiography was lower among elderly patients than among younger patients (52% vs 61%), but the sensitivity was higher (87% vs 75%). In contrast, both the sensitivity and the specificity of dipyridamole thallium scintigraphy were similar in the two age groups (86% vs 87% and 79% vs 74%). Among patients with myocardial infarction and a positive exercise test, reversible defects were equally common in the two age groups (60% vs 58%). The reversible defects were in areas remote from the area of infarction in half of the patients in both groups. Among patients with negative exercise-test results, reversible defects were more common in elderly patients than in younger patients (57% vs 38%). The reversible defects were in the infarcted area in 71% and 79% of these patients, respectively. These results indicate that dipyridamole thallium scintigraphy is a sensitive and specific method for detecting CAD, independent of age. It is particularly useful in identifying myocardial viability in the infarcted area. PMID- 9396317 TI - [Helicobacter pylori infection and endoscopic appearance of the gastric mucosa in elderly patients with peptic ulcer]. AB - I examined the characteristics of Helicobacter pylori (H. pylori) infection in elderly patients with peptic ulcer and its relation to the endoscopic appearance of the gastric mucosa. 1) Infection with H. pylori was more common in middle-aged patients (those over 40 and younger than 59 years old, 80.4%) than in younger patients (those less than 39 years old, 63.0%). Elderly patients were less likely than younger patients to be infected (60's: 77.7%, 70's: 70.8%, over 80 years old: 65.8%). The percentage was higher in men than in women, in all age groups. 2) Oshima's classification was used to divide the patients into 5 groups, according to the endoscopic appearance of blood vessels of the gastric mucosa. Infection was found in 71.7% of the patients without atrophy, in 86.3% of those with mild atrophy, and in 88.9% of those with moderate atrophy. In contrast, infection was found in only 78.4% of the patients with severe atrophy. Similar results were found in patients with peptic ulcer and in subjects with no lesion except atrophic gastritis. 4) The percentage of patients with gastric ulcer disease who had atrophic gastric mucosa was higher in those with ulcers above the middle of the stomach (46.3%) than in those with ulcers in the antrum (30.2%, p < 0.05). Almost all patients with gastric ulcers in the lower part of the stomach and in the angulus were found to be infected with H. pylori (93.3% and 94.0%, respectively). The percentage of patients with ulcers low in the stomach who were infected was lower (59.4%). All of the location-related differences in infection were significant (p < 0.001). PMID- 9396318 TI - [Evaluation by specialists of the Guidelines on Treatment of Hypertension in the Elderly--the second questionnaire survey]. AB - The Guidelines on Treatment of Hypertension in the Elderly, 1995 was published by the Research Group for Guidelines on Treatment of Hypertension in the Elderly Comprehensive Research Projects on Aging and Health, the Ministry of Health and Welfare. To assess the guidelines, and to further investigate the changes in the therapeutic policy for hypertension in the elderly, we mailed a questionnaire to 133 Japanese hypertension specialists who had replied to the first mailed questionnaire in 1993. We received 102 replies (77%). Overall, the guidelines were scored 4.3/5.0. More than 95% of the specialists agreed with levels of blood pressure (BP) indicative for treatment, as well as the goal of BP control. The guidelines propose that the rate of increase of the drug dose should be very slow, at least every four weeks, and that the target BP be reached after two months. The majority of the specialists agreed with this. However, 10% of them preferred to follow patients every two weeks. The guidelines propose long-acting Ca antagonists and ACE inhibitors as the first choice of drug for the treatment of uncomplicated hypertension in the elderly. Two-thirds of the specialists agreed. However, 17% of specialists proposed adding diuretics or beta-blockers to the first line therapy. Ten specialists (10%) expressed concerns about Ca antagonists and three (3%) insisted on withdrowing them from the first line of drugs. In the guidelines, alpha and beta blockers are designated as relatively contraindicated in elderly patients with hypertension, but half the specialists answered that these drugs can be used safely in elderly patients. These findings indicate that the therapeutic policy of Japanese specialists in hypertension in the elderly has not changed substantially for three years. PMID- 9396319 TI - [Deterrent factors in rehabilitation training following cerebral vascular disease -comparison of elderly and non-elderly patients]. AB - The purpose of the present study was to clarify the deterrent factors for rehabilitation training (rehab) of chronic cerebral vascular disease (CVD) patients, and also to evaluate the influence of age on these factors. Sixty-five CVD impatients with sequelae treated at the Cerebral Vascular Center of Nanasawa Hospital were included in the study. Patients were classified into two groups using the Barthel index score: good effect group (n = 21) or no effect group (n = 22). The following factors were compared between the two groups in order to investigate which factor most affects the results of rehab: age, sex, the site of brain damage, extent of motor paralysis, character (Type A character or not), aphasia, hemispacial neglect, depression, and positive attitude toward training. A possible association between depression, and the site of brain damage and Type A character was investigated. Also, the difference in mood disorders was compared between elderly and non-elderly stroke patients. In the elderly group, hemispacial neglect, a negative attitude toward training, and depression all adversely affected the outcome of the rehab. In the non-elderly group, aging, hemispacial neglect, and a negative attitude toward training influenced the effect of the rehab, but there was no correlation with depression. Depression was seen in 64% of the patients (38/59). Of the 38 patients in a depressed state, 24 (63%) had right hemisphere brain damage, 13 (34%) had left hemisphere brain damage, and 1 (3%) had brain stem damage. Twenty-seven of the 38 depressed patients (71%) were Type A character, significantly more than in the non depressed group (92/21, 43%). In addition, 14 of the 27 Type A patients were aged over 65 years (52%), which was more than in the non-depression group (11/38, 29%). PMID- 9396320 TI - [Investigation of autopsy cases of patients who had endoscopic injection sclerotherapy performed, classified by their age at the time of initial therapy]. AB - We performed endoscopic injection sclerotherapy (EIS) for esophageal varices on approximately 214 patients between October 1981 and July 1994 at our hospital. Of the 214 patients 114 have died, and we divided them into two groups according to their age when EIS was first performed: (i) group 1, less than 70 years old; and (ii) group 2, more than 70 years old. We investigated the efficacy of EIS for the group 2 patients with esophageal varices by comparing the two groups. EIS was considered effective in the group 2 patients because there was no difference between the two groups in the period of observation after EIS, but the time to re therapy in the autopsy cases of this age group was significantly less. As a result of investigating the surgical outcome and the direct cause of death, it was suggested that; in future, prevention of death by hepatocellular carcinoma and hepatic failure was necessary for both groups. PMID- 9396321 TI - [Ataxic neuropathy in the elderly--clinicopathological study of six cases]. AB - The combination of ataxia with peripheral neuropathy (ataxic neuropathy) is rare. Six elderly patients with peripheral neuropathy who developed ataxia were studied. Of the peripheral neuropathies, ataxic neuropathy was significantly more frequent in patients aged more than 65 years compared with younger patients. Ataxic neuropathy was associated with carcinoma (2 cases), Sjogren's syndrome (1 case), chronic inflammatory demyelinating polyradiculoneuropathy (CIDP, 1 case) and chronic idiopathic ataxic neuropathy (2 cases). The two cases of carcinomatous neuropathy initially showed ataxia, which preceded detection of the carcinomatous lesions in the lung by approximately 1 year. The study cases had many clinical features in common. In the nerve conduction study, sural nerve action potential could not be measured in five of the cases; sural nerve biopsy revealed a decreased density of myelinated fibers in all cases. In particular, the large myelinated fibers were markedly decreased. These findings were common, regardless of the underlying disease, except in the case of CIDP in which there was only a slight decrease in the number of large myelinated fibers. Differential diagnosis based on the clinicopathological features was difficult. Therefore, in cases of ataxic neuropathy, systemic evaluation is necessary to rule out the possibility of carcinoma or various systemic diseases, especially in elderly patients. PMID- 9396322 TI - [Relationship between knee pain and X-ray findings of osteoarthritis with reference to bone mineral density measured by computed X-ray densitometry]. AB - We studied the relation between performance of activities of daily living and X ray findings of osteoarthritis of the knee in aged persons. We made Covariance Structure Analysis models of knee pain, X-ray findings and bone mineral density as measured by computed X-ray densitometry. The subjects were 257 women aged from 47 to 88 years who were outpatients at an orthopedic clinic. The best-fit model indicated that loss of bone mineral density in metacarpals was associated with X ray findings of knee-joint degeneration, as well as with knee pain. No relationship was found between knee-joint degeneration and knee pain. These results suggest that bone mineral density should be taken into consideration when interpreting X-ray findings of knee pain. This model incorporates the effects of degeneration of the subchondral bone. In summary, we should use measurements of bone mineral density along with X-ray findings for the diagnosis and treatment of knee osteoarthritis. We may be able to precisely predict knee pain caused by osteoarthritis, by analysis of a model that includes a lesion in joint cartilage. PMID- 9396323 TI - [Relationship between cognitive deficits, behavioral disturbances and falls in patients with dementia]. AB - Prospective study was conducted in order to investigate the relationship between cognitive and behavioral disturbances and falls in dementia. Falls and fall related fractures were studied in 154 ambulatory patients who were admitted to a geriatric intermediate nursing facility with a diagnosis of Alzheimer's type dementia (DAT) or mixed Alzheimer and vascular dementia (MIX). The Mini-Mental State (MMS) was used for the evaluation of cognitive status, and the Dementia Behavior Disturbance Scale (DBD) was used as a behavioral parameter. In order to evaluate problematic behavior in walking, a derivative scale (DBD-W) was developed from the DBD by choosing three items relevant to wandering. During the 3 months of observation, 61 patients fell more than once; 15 of them experienced fractures. The mean MMS scores were higher in non fallers, fallers who did not sustain fractures and those complicated with fractures, in that order, and the difference between the non fallers and fallers with fractures was statistically significant. When the subjects were divided into three subgroups on the basis of the MMS scores, there was a trend toward a higher ratio of fallers or those complicated with fractures in the subgroup with the lower MMS score. The DBD and DBD-W scores were not associated with falls or fractures, although fallers had a slightly higher DBD or DBD-W score than non fallers. These findings suggest that the risk of falling or fracture becomes higher with the advance of cognitive impairment in institutionalized ambulatory patients with DAT or MIX. The findings also suggest that behavioral disturbances are not necessarily associated with falls or fractures. PMID- 9396324 TI - [A case of polymyalgia rheumatica with swelling and pitting edema of the distal lower extremities]. AB - We report a case of an 82-year-old woman with polymyalgia rheumatica (PMR) associated with swelling and pitting edema of the lower extremities. The patient had been previously admitted because of PMR in 1990, but there was no history of swollen extremities. In July 1996, at another hospital, she was again diagnosed as having PMR on the basis of pain in the neck, shoulders and lower back. Administration of prednisolone was followed by improvement of the symptoms. Four months later, similar pain recurred and swelling of the lower extremities was noted. On admission, the erythrocyte sedimentation rate was 86 mm/h, and C reactive protein was 15.5 mg/dl. Reviewing the previous treatment, it was ascertained that her clinical deterioration was due to premature reduction of the steroid dosage. The cause of the swelling of the lower extremities was unlikely to be heart, liver, kidney or endocrine disease. Prednisolone was increased from 2.5 mg to 10 mg daily with marked improvement in all the symptoms including the swelling and pitting edema. In 1996, a study reported distal extremity swelling with pitting edema as a manifestation of PMR, which mostly developed concurrently with proximal symptoms or during relapses of PMR. The swelling responded poorly to non-steroidal antiinflammatory drugs but promptly to corticosteroids. The distal swelling was reported to be tenosynovitis and synovitis of the surrounding structures. The present case appears similar to that report. More studies of PMR need to be done. PMID- 9396325 TI - [Septicemia due to methicillin-resistant Staphylococcus aureus from chronic prurigo in an elderly woman]. AB - An 80-year-old woman being treated with anti-hypertensive drugs developed eruption and itching of the skin. High fever and lymph node enlargement subsequently developed in spite of discontinuing all antihypertensive drugs, and she was admitted to our hospital. At the initial examination, multiple papules were noted over the entire body, and the skin showed thickening and lichenification with scratch marks. There was also generalized enlargement of the superficial lymph nodes. From these findings, her condition was diagnosed as chronic prurigo due to drug allergy. Laboratory tests showed inflammatory findings, anemia and a high serum level of IgE. Analysis of the surface marker of peripheral lymphocytes revealed no abnormalities. Bacteriologic cultures of blood revealed methicillin-resistant Staphylococcus aureus (MRSA). Histologic examination of the lymph nodes revealed chronic reactive lymphadenitis with a follicular pattern. She was strongly suspected of having MRSA septicemia, and so combination chemotherapy with vancomycin, minocycline and cefoperazone/sulbactam was started. However, 1 month after initiation of chemotherapy, the low-grade fever, eruption and moderate inflammatory findings persisted, and culture of the eruptions revealed MRSA. The prurigo was therefore considered to be the source of the septicemia, and daily application of diflucortolone ointment containing 3% acetic acid was started. Thereafter, the clinical and laboratory findings showed a rapid improvement. MRSA infections usually occur in compromised patients who are receiving antibiotics during prolonged hospitalization. The present case, who did not have any underlying disease, indicates that old-age is also an important factor for the development of MRSA septicemia. PMID- 9396326 TI - [Changes in serum anti-Helicobacter pylori IgG antibody, pepsinogen I, and pepsinogen II after eradication therapy of Helicobacter pylori]. AB - To investigate the changes in serum anti-Helicobacter pylori IgG antibody (HP Ab), pepsinogen I (PG I), pepsinogen II (PG II), and pepsinogen I/II ratio (PG I/II) after eradication therapy of Helicobacter pylori (HP), we studied 78 patients with HP-positive peptic diseases. They received combination therapy (proton pump inhibitor + amoxicillin: n = 17, proton pump inhibitor + amoxicillin + clarithromycin: n = 61). In the 68 patients in whom HP was eradicated, HP Ab, PG I, and PG II decreased and PG I/II increased significantly after eradication. Especially, the decrease in PG II and the increase in PG I/II were rapid and remarkable, found 2 months after the beginning of eradication therapy, and then continued. On the other hand, in the patients in whom HP was not eradicated, HP Ab and PG I/II did not change significantly, while PG I and PG II temporarily increased at the end of administration of proton pump inhibitor. In conclusion, it seems that the measurement of PG II and PG I/II is useful for the early detection of HP eradication. PMID- 9396327 TI - [Percutaneous ethanol injection therapy by ethanol mixed with CO2 microbubble for hepatocellular carcinoma]. AB - One of the shortcomings of percutaneous ethanol injection therapy (PEIT) for hepatocellular carcinoma (HCC) is that many sessions are necessary to accomplish the treatment. This may be caused by which the ultrasonography (US) image does not reflect correctly to the kinetics of injected ethanol into HCC nodule. It is considered that number of treatment sessions are able to be reduced if we just enough injected labelled ethanol under US into HCC nodule. Therefore, we tried PEIT by ethanol mixed with CO2 microbubble (CO2 ethanol). The injected CO2 ethanol was aquired as hyperechoic image without strong acoustic shadow to the end of injection. Consequently we could reduce the number of treatment sessions to almost 1 for lesions < or = 3 cm in diameter and markedly reduce total dose of injected ethanol. The detectable rate of CO2 ethanol leaked out HCC nodule was high. No serious complication occurred. There have been only 1 lesion of local recurrence and no case of intrahepatic and peritoneal dissemination for 11.5 months on average of observation after PEIT by CO2 ethanol (CO2PEIT). These findings suggest that CO2PEIT is useful method for reducing the number of treatment sessions and total dose of injected ethanol, moreover preventing complication by ethanol leakage. PMID- 9396328 TI - [Histopathological study in models of chronic pancreatitis]. AB - Histopathological findings were examined in the models of chronic pancreatitis. Using mongrel dogs, we prepared a control group (group C), a chronic ischemic group (group I), an alcohol administration group (group A), a duct obstruction group (group O), and an alcohol + obstruction group (group AO). Group I showed severe inflammatory cell infiltration, fibrosis, fat replacement and loss of acinar cells. Group A showed no change. In group O, mild periductal fibrosis was recognized. Group AO showed moderate inter-lobular fibrosis and inflammatory cell infiltration, resembling those of human chronic alcoholic pancreatitis. CONCLUSION: 1) Histological findings of chronic ischemic group is severer than that of group O and AO. 2) The model of alcohol administration with incomplete duct obstruction is a useful model of human chronic alcoholic pancreatitis. PMID- 9396329 TI - [A case of carbohydrate antigen 19-9 producing gastric mucinous adenocarcinoma with the appearance of a submucosal tumor]. PMID- 9396330 TI - [A case of PTH-rP producing gastric cancer with extragastric growth]. PMID- 9396331 TI - [A case of pseudomyxoma peritonei of appendiceal origin that metastasized to the spleen and complicated secondary systemic amyloidosis]. PMID- 9396332 TI - [A case of Crohn's disease associated with pyoderma gangrenosum]. PMID- 9396333 TI - [A case report of primary intestinal lymphangiectesia successfully treated with low fat diet]. PMID- 9396334 TI - [A case report of ulcerative colitis complicated with protein losing enteropathy and colon cancer in a young female]. PMID- 9396335 TI - [A case of refractory diarrhea treated with somatostatin analogue]. PMID- 9396336 TI - [A case of Meckel's diverticulum caused intestinal obstruction of loop formation and strunglated intestine by adhesion]. PMID- 9396337 TI - [A case of drug induced hepatitis and interstitial pneumonia caused by a herbal drug, Dai-saiko-to]. PMID- 9396338 TI - [A case of hemorrhagic retention cyst of the pancreas]. PMID- 9396339 TI - [Changes in serum lipoprotein fraction in patients with chronic hepatitis C during interferon beta treatment]. PMID- 9396341 TI - [Etiological analysis of thrombophilia]. AB - We have established a system for etiological analysis of thrombophilia which includes assays of antithrombin III, protein C, protein S, plasminogen, fibrinogen, heparin cofactor II and lupus anticoagulants as well as gene analysis. The analysis conducted on 115 patients with venous thrombosis, arterial thrombosis and small vessel thrombosis revealed that forty-one patients(36% of the examined patients) were accompanied with decreased activities of protein S, protein C, antithrombin III and plasminogen. Eleven candidate causal mutations were found by gene analysis. These studies indicate that a comprehensive examination is instrumental in identifying and confirming the etiology in patients with thrombophilia. PMID- 9396343 TI - [Apoptosis in Hashimoto's thyroiditis]. AB - It has been suggested that apoptosis plays a pivotal role in the pathogenesis of autoimmune diseases. In Hashimoto's thyroiditis which is a typical organ-specific autoimmune disease, Fas-FasL-mediated apoptosis has been demonstrated as the mechanism of follicular epithelial cell death in which Fas is expressed by IL-1 beta stimulation of FasL is constitutively expressed on follicular epithelial cells. The processes involved in this finding and some questions concerning epithelial cell death are presented. The thyroid tissue of Hashimoto's thyroiditis was examined for DNA fragmentation of follicular epithelial cells by the TUNEL method. DNA fragmentation was observed more frequently on thyroid follicles in the area adjacent to lymphoid cell follicles than on those in the central area. Electron microscopic study supported the results of TUNEL study. Immunohistochemical study on Fas and FasL expression on follicular epithelial cells of various thyroid diseases showed that Fas and FasL were strongly expressed on follicular epithelial cells in Hashimoto's thyroiditis and thyroid cancer. Epithelial cells of patients with Graves' disease and adenomatous goiter, however, were scarcely stained. Fas and FasL expression on follicular epithelial cells were well correlated. In vitro study on follicular epithelial cells clarified that FasL was constitutively expressed on epithelial cells not only in Hashimoto's thyroiditis but also in nontoxic goiter. Fas expression was induced by IL-1 beta stimulation. IL-1 beta stimulation also brought about apoptosis of epithelial cells and epithelial cells killed Fas-positive target cells. Therefore, it was concluded that FasL expressed constitutively on follicular epithelial cells interacts with Fas on epithelial cells expressed by IL-1 beta stimulation to induce apoptosis of epithelial cells. PMID- 9396342 TI - [Support system for diagnosing hematologic malignancies]. AB - We established a Southern blot hybridization using a DIG-labeled probe to detect monoclonal integration of HTLV-1 proviral genome. DIG was labeled by the PCR method and this probe was as sensitive as the 32P-labeled probe and able to detect only 1.6% of ATL cells. The clinical diagnoses of 44 patients with monoclonal band(s) were all ATL. In contrast, the clinical diagnoses of 39 patients without monoclonal band(s) were diseases other than ATL. We also performed a long PCR of HTLV-1 to characterize the integrated provirus. The method allowed us to find a defective provirus, which was frequently observed in aggressive forms of ATL; 14 of the 18 patients with acute type(78%), 6 of the 9 patients with lymphoma type(67%), and 2 of the 12 patients with chronic type(17%) had the defective provirus. We established a simultaneous PCR for each region of HTLV-1 for further examination of the defective provirus. To detect the monoclonality of IgH gene rearrangement of B-cells, we performed PCR according to the method described. None of 13 patients with T-lymphoproliferative disorders showed a monoclonal band. In contrast, 12 of 13 patients with CLL(92%), 22 of 25 patients with common ALL(88%), 18 of 24 patients with B-lymphoma(75%), and 3 of 3 patients with hairy cell leukemia(100%) showed a monoclonal band. We are now expanding this kind support system for the clinical diagnosis at a molecular biology level in the central laboratory. PMID- 9396344 TI - [Molecular pathomechanism of HTLV-I infectious diseases]. AB - Human T-cell lymphotropic virus type I(HTLV-I) is a type C retrovirus, closely associated with adult T-cell leukemia. In the past few years, studies have revealed an association between the virus and disorders of organ systems including myelopathy, polymyositis, alveolitis, and Sjogren's syndrome. In addition, we have proposed that HTLV-I-associated proliferative changes in nonlymphocyte synovial cells are termed HTLV-I-associated arthropathy(HAAP). To clarify the pathologic association of HTLV-I with this arthropathy, we attempted to detect HTLV-I proviral DNA in synovial tissue. Proviral HTLV-I DNA was detected not only in peripheral blood mononuclear cells but also in fresh synovial tissue cells and lymphocyte-depleted cultured synovial cells. We also detected mRNA for HTLV-I tax/rex in cultured synovial cells by reverse transcription polymerase chain reaction. Moreover, induction of chronic inflammatory arthropathy in mice transgenic for HTLV-I tax gene strongly suggested the pathogenic mechanism of HAAP. Histologic findings of affected joints in mice showed erosions of bones and pannus-like granulomtous change with infiltration of mononuclear cells. Thus, this novel mechanism might explain synovial proliferation caused by HTLV-I. Tax-expressing transgenic mouse lines also demonstrated that tax itself could serve as an oncogne in fibroblastic cells. Tumors occurred in 100% of the mice with reproducible time periods after wounding. We established cell lines, which expressed high levels of c-fos, c-myc, myb, PDGF, TGF-beta, Zif, and IL-6. Antisense ablation of the p65 subunits of NF kappa B profoundly inhibited tumor growth in vitro with no apparent affect on the growth of normal cells. These studies were successfully extended to tax transgenic animals. Intraperitoneal injections of NF-kappa B p65 antisense at the 40 micrograms/g weight dose led to growth arrest after 7 days, and apparent involution after 20 days of treatment. We think activation of NF-kappa B by Tax is important for tumor progression. This paper supports the importance of analyzing molecular pathomechanisms. PMID- 9396345 TI - [Determination of neutrophil function by measuring superoxide production with whole blood flow cytometry]. AB - The function of neutrophil can be evaluated by measuring oxidative metabolism using chemiluminescence, tetrazolium dye reduction or the others. Those results are not always satisfactory which would be caused by subtle difference in each preparation of the reagents and the lack of reproducibility. Recently, flow cytometric procedures for semi-quantitating superoxide production in neutrophils have been developed to evaluate their function. This procedure, which requires only small amount of whole blood, can easily and rapidly yield reproducible and reliable data. In this study, we optimized analytical conditions and then determined reference interval to evaluate neutrophil function of patients with various disorders. Optimal concentrations and incubation times of DCFH-DA and PMA were 5 mumol/l for 15 minutes and 25 micrograms/ml for 20 minutes, respectively. Production of superoxide in neutrophil was represented by relative fluorescence intensity(RFI) with assay coefficient of variance(CV) of 4.0-11.1%. Neutrophils had to be examined within 2 hours after venipuncture to obtain reliable data. Reference interval was determined as 170.4 +/- 58.7(mean +/- SD) RFI. Neutrophil function of patients with neutropenia, paroxysmal nocturnal hemoglobinuria(PNH), renal failure, systemic lupus erythematosus(SLE), myeloperoxidase deficiency, myelodysplastic syndrome(MDS), and diabetes mellitus were within the reference interval as evaluated by this method. Only neutrophils of chronic granulomatous disease, which is known to give clearly low superoxide production, showed actually decreased value. These results indicate that this procedure would be clinically useful for diagnosis of patient with impaired neutrophil function. PMID- 9396346 TI - [Determination of uric acid in scalp hair for non-invasive estimation of uricemic control in hyperuricemia]. AB - Uric acid in blood has been widely accepted as a reliable indicator of hyperuricemia and gout, and its assay method has been established. In the present study, we developed a simple and non-invasive rapid method for the determination of uric acid in hair. Concentration(nmol/mg hair) of uric acid extracted from 10 20 mg of hair(95% < extractability; 0.1N KOH, 37 degrees C, 2 hr) was determined by an enzymatic method using uricase. The concentration of uric acid(nmol/mg hair, mean +/- SD: 0.489 +/- 0.157, n = 16) in hair from hyperuricemic patients was significantly higher than that(0.258 +/- 0.107, n = 8) from healthy volunteers(p < 0.01). Within-run and between-day precisions(reproducibilities, CVs) for the assay were 9.6-10.3%(n = 10 each) and 11.6-16.3%(n = 7 each), respectively. The concentration(y, nmol/mg hair) of uric acid in hair correlated well with that in blood(x, g/l): y = 8.770x-0.123(r = 0.746, Syx = 0.122, n = 23). Changes in the concentration of uric acid in hair of hyperuricemic patient treated with allopurinol paralleled to those in blood. In conclusion, it was confirmed that the concentration of uric acid in hair reflected that in blood, suggesting that measuring uric acid in hair can be available for the metabolic control in hyperuricemia. PMID- 9396347 TI - [Angiotensin I-converting enzyme gene polymorphism and renal disease]. AB - ACE inhibitor is known to have a therapeutic efficacy in renal diseases by reducing proteinuria and maintaining renal function. However, the relationship between ACE gene polymorphism and renal disease has not been fully elucidated. In this study, a 287 base pair(bp) I/D polymorphism of the ACE gene was examined with polymerase chain reaction(PCR) in 100 healthy subjects, 34 patients with chronic glomerulonephritis(CGN), 29 with chronic renal failure(CRF) and 25 with diabetes mellitus(DM) with(13) and without(12) nephropathy. We also measured serum ACE activity of these patients. ACE genotype and derived allele frequencies in each disease group did not differ significantly from those in healthy subjects. In all disease groups, values of serum ACE activity were higher in genotype DD than in genotype II. These findings suggest no significant association between I/D polymorphism of the ACE gene and renal disease. Further studies are needed to clarify these findings, considering renal function and type of renal disease. PMID- 9396348 TI - [Establishment of the optimal conditions for detecting anti-M2 by western blotting in primary biliary cirrhosis]. AB - Anti-mitochondrial antibodies(AMA) are serodiagnostic markers for primary biliary cirrhosis(PBC) but heterogenous in antigen molecules which they recognize. A disease-specific AMA for PBC is anti-M2. The conventional examination methods are indirect immunofluorescence and ELISA. However, there are some problems in specificity, because the antigen preparations used are crude. Thus, analysis with Western-blotting(W-B) is needed, because it allows the identification of a molecule which the antibody reacts with. In this report, we established the optimal conditions for detecting anti-M2 with W-B in PBC. As antigen, we used mitochondrial fractions derived from beef hearts. Because a positive band at 74 kDa became negative after absorbing sera with PDH purified from porcine hearts, this band corresponded to major antigeneity of anti-M2. Titration experiments with SDS-PAGE showed that the optimal concentration of this antigen preparation for loading is 0.04 mg/ml. We also performed titration experiments to determine the optimal dilutions for second antibodies and serum samples. The results showed that the optimal dilution for second antibodies, anti-IgG and anti-IgM, were 1:3000 and 1:1000, respectively. The optimal dilution for serum samples was shown to be 1:10(2). Moreover, the W-B technique gave a positive result even for sera from AMA-negative PBC patients which had tested negative with conventional methods, if undiluted sera were examined or if anti-IgG or anti-IgM was used as a second antibody. Thus, the W-B technique is more sensitive than conventional methods of analysis. Based on these results, we will be able to detect anti-M2 with maximum efficiency, thus improving our ability to study the relationship between anti-M2 and the pathological conditions in PBC. PMID- 9396349 TI - [Western blot analysis of anti-M2 antibodies in anti-mitochondrial antibody negative primary biliary cirrhosis]. AB - One variety of anti-mitochondrial antibody(AMA) is characteristically found in sera from patients with primary biliary cirrhosis(PBC). The major target antigens of this type of AMA are M2s. It is well known, however, that AMA-negative PBC also exists. An alternative disease concept, called autoimmune cholangiopathy, recently has been advocated. This new concept is defined by the following criteria: 1)the failure to detect AMA and anti-M2, 2)the detection of a diffuse type of anti-nuclear antibody and anti-smooth muscle antibody, 3)pathological findings compatible with PBC, and 4)the effectiveness of prednisolone. However, the difference between AMA-negative PBC and autoimmune cholangiopathy is controversial. Therefore, we analyzed antibodies to four major M2 proteins with Western blotting in 34 cases of immunofluorescent AMA-negative PBC. In 31(91.2%) of these 34 AMA-negative sera, antibodies to at least one of these four major M2 proteins was detected. In serum samples from 34 control patients with AMA positive PBC, antibodies to at least one of these four proteins were detected in all cases. In addition, we studied the frequency of cases which satisfied the serological criteria of autoimmune cholangiopathy. In only one(0.7%) of 141 cases was the serological criteria met. We conclude that to clarify the serological differences between autoimmune cholangiopathy and AMA-negative PBC, the analysis of M2 proteins by Western blotting is essential. PMID- 9396350 TI - [Serum levels of soluble tumor necrosis factor receptors as markers for disease progression of human immunodeficiency virus infection]. AB - We studied whether the serum levels of soluble tumor necrosis factor receptor(sTNFR) type I or II correlate with clinical progression of human immunodeficiency virus type 1(HIV-1) infection. Serum levels of sTNFR type I and II were measured by an enzyme-linked immunosorbent assay in sixty five patients with HIV-1 infected hemophiliacs and 10 healthy controls. In a longitudinal study, we assessed whether the decline of CD4+ lymphocyte counts were associated with increased serum concentrations of sTNFRs. Elevated serum concentrations of sTNFRs were found among the HIV-1 infected patients and higher in patients with advanced clinical stage. We noticed there were two distinct patient groups in change of CD4+ lymphocyte count when twenty-eight patients were followed retrospectively for a median period of 65 months. 14 patients represented stable CD4+ lymphocyte counts, but another 14 patients had more than 40% decrease of CD4+ lymphocyte counts over the course of this study. Serum sTNFR type II levels were 3.49 +/- 0.71 to 3.11 +/- 0.31 ng/ml in the former group, and 4.88 +/- 0.91 to 4.26 +/- 0.71 ng/ml in the latter group during the study. Significantly higher levels of sTNFR type II were already revealed at early time in the latter group. There was, however, no significant difference in the level of sTNFR type I between the two. These results suggest that serum levels of sTNFR type II provided useful information as a predictor of disease progression related to the decline of CD4+ lymphocyte counts. PMID- 9396351 TI - [Analysis of genome-type, serovar and antibiotic susceptibility of Pseudomonas aeruginosa isolated in Beijing Hospital China in 1991 to 1993]. AB - We analyzed the in vitro antibiotic susceptibility pattern and serovar for 192 strains of Pseudomonas aeruginosa isolated at Beijing Hospital(China) from October 1991 to October 1993. The frequency of resistant strains was high, more than 15%, for ceftazidime, cefsulodin, cefoperazone, aztreonam, gentamicin, tobramycin, tosufloxacin, ofloxacin and fosfomycin. The incidence of resistants against piperacillin and imipenem was significantly low. Among the 192 strains, 16 were designated as being multi-drug resistant strains(i.e.; resistant to more than 8 drugs out of 11 drugs), and all of the multi-drug resistant strains were isolated from inpatients. Predominant serovar of 192 strains were 60(31.3%) for G, 28(14.7%) for E, and 24(12.5%) for I. Multi-drug resistant strains have no characteristic serovar. Restriction enzyme SpeI digestion analysis(using pulse field electrophoresis) of P. aeruginosa-genome yielded several common patterns, and was shown to be useful for tracing the route of nosocomial infection. Further, isolates with closely related genome type, in which the size of one digested band was different, showed a different minimum inhibitory concentration of fosfomycin in one genome type or new quinolones in the other genome type. Analysis of genome type and antibiotic susceptibility pattern may exhibit the antibiotic resistant gene. PMID- 9396352 TI - Differences in molar absorptivity of 4-NP with the reaction solution and apparatus affect ALP measurement. AB - We examined the differences in molar absorptivity of 4-NP obtained using different kits for ALP measurement and different instruments. The apparent molar absorptivity of 4-NP in the same reaction solution determined by six different instruments was 15.98, 16.72, 16.06, 17.00, 16.27, 17.62 and that using four different reaction solution kits for ALP with the same instrument was 16.90, 17.38, 17.72, 16.11. We measured ALP in three serum samples with six instruments using the same kit and in twelve serum samples with the same instrument using four kits. ALP activities measured using the same molar absorptivity value differed with the instrument(p < 0.01). However, those measured using the apparent molar absorptivity value for each instrument revealed no significant differences(p > 0.05). In conclusion, we suggest that standard material should be contained in each kit for enzyme measurement and the apparent epsilon for each kit and instrument should be obtained to minimize the systematic error caused by using the same epsilon in different laboratories. PMID- 9396353 TI - [Cytokine-secreting cells in relapsing multiple sclerosis patients treated with high-dose intravenous methylprednisolone]. AB - Interleukin-2 (IL-2), interleukin-2 receptor (IL-2R), interferon gamma (INF gamma), interleukin-4 (IL-4) secreting peripheral blood cells were enumerated by enzyme-linked immunospot (ELISPOT) assay in 9 relapsing multiple sclerosis (MS) patients treated with high-dose intravenous methylprednisolone (MP), in 7 MS patients with remission,and in 9 controls. IL-2, IL-2R, INF gamma, IL-4 secreting cells were significantly higher before MP therapy in relapsing MS patients as compared with after MP therapy in relapsing MS patients, with MS patients in remission, and with controls. IL-2, INF gamma secreting cells before MP in relapsing MS patients and those in remission significantly increased in response to myelin basic protein (MBP) 4-14, 68-84 and proteolipid protein (PLP). INF gamma secreting cells after MP therapy in relapsing MS patients significantly increased in response to MBP 4-14, 68-84, and PLP. IL-2R secreting cells in MS patients in remission significantly increased in response to MBP 4-14, 68-84, and PLP and were significantly higher after MP therapy in relapsing MS patients and in controls. This fact suggested that there was a systemic T-cell response to myelin antigens in MS patients in remission. The use of ELISPOT to investigate cytokine-secreting cells may play a role in the evaluation of clinical activity during treatment with high-dose intra-venous MP in relapsing MS patients. PMID- 9396354 TI - [A study of MRI and clinical neurology in acute cerebellar infarcts]. AB - We studied clinical manifestations of sixteen patients with cerebellar infarcts diagnosed by MRI. In fourteen of them, the stroke developed abruptly with vertigo, which continued for several days. At the early stage of illness, ataxia was obscure. But after vertigo and nausea disappeared, nine cases showed truncal ataxia, while limb ataxia was found in only five. Their vertigo was rotatory and aggravated by head movement. Gaze-evoked nystagmus was observed in only 5 cases. Four patients preferred to take unilateral posture since they experienced less vertigo. The side of their lesions was the lower side of their posture. Limb ataxia was more frequent in SCA-involving cases than in SCA-non involving cases (3 out of 6 vs 2 out of 10, respectively). On the other hand, headache was more frequent in PICA-involving cases than in PICA-non-involving cases (6 out of 11 vs 1 out of 5, respectively). Ataxic gait was seen more in medial branch-involving cases than medial branch non-involving cases (5 out of 6 vs 4 out of 10, respectively). One patient died due to obstructive hydrocephalus. PMID- 9396355 TI - [Efficacy of TRH-T for spinocerebellar degeneration--the relation between clinical features and effect of TRH therapy]. AB - Thyrotropin releasing hormone (TRH) therapy has been frequently attempted for the treatment of spinocerebellar degeneration (SCD) and its efficacy has been confirmed. However, effectiveness is considered to differ depending on disease type, severity and the method of evaluating clinical improvement. We investigated the efficacy of thyrotropin releasing hormone-tartrate (TRH-T) in 23 patients with SCD consisting of cerebellar form (cortical cerebellar atrophy (CCA) and hereditary cortical cerebellar atrophy (H-CCA)), and multiple system form (multiple system atrophy (MSA) and hereditary olivopontocerebellar atrophy (H OPCA)). TRH-T, 2 mg per day, was given intravenously for 20 days. The effect of TRH therapy was evaluated by assessing changes in balance function while lying, sitting, standing and walking, that may reflect the movement functions in active daily life (ADL) for the patients with SCD. The speech function was also evaluated qualitatively using acoustic analysis. The amine metabolites (HVA and 5 HIAA) in cerebrospinal fluid possibly reflecting the noradrenaline and serotonin metabolism in the central nervous system were measured before and after treatment. Although mild or moderate improvement of the balance function during the course of TRH therapy was seen in 16 of the 23 patients, patients with cerebellar forms (CCA and H-CCA) improved significantly as compared to patients with MSA. The effect persisted for a long time (mean; 3.8 months) after TRH therapy in nine of the 16 patients, and eight (88.9%) of the nine had the cerebellar form of SCD. The levels of HVA and 5-HIAA in CSF also increased in patients with CCA as compared to patients with MSA and H-OPCA. The disease severity before the treatment in 14 (87.5%) of 16 patients who showed improvement of balance functions by TRH therapy was mild or moderate; possible of walking without support, or occasionally with support. Considering these results together, TRH therapy may be effective in patients with the cerebellar form of SCD, whose illness severity is mild, and may be recommended for support of ADL in patients with the cerebellar form of SCD. PMID- 9396356 TI - [Bilateral vertebral artery occlusion]. AB - We studied 9 patients with bilateral vertebral artery occlusion (BVAO). BVAO was confirmed using angiography in order to clarify its clinical feature, mechanism, and long term prognosis. Three patients showed bilateral intra-cranial occlusion, 3 bilateral extra-cranial occlusion, and 3 intra and extra-cranial occlusion. The basilar artery was fed by the posterior communicating artery in 8 out of 9 patients. In one of the 8, reconstitution of the thyrocervical artery was seen. We divided the patients into 4 groups according to MRI findings, as follows: Group 1 with no abnormal finding on MRI (N = 2); Group 2 with deep pontine infarcts and non-territorial small cerebellar infarcts (N = 2); Group 3 with extended pontine infarcts (N = 3); and Group 4 with cerebellar cortical artery infarcts, deep pontine infarcts, and non-territorial small cerebellar infarcts (N = 2). Transient episodes were seen in all patients, 8 patients out of 9 had vertigo/dizziness, 3 tinnitus, 2 diplopia, 2 headache, 2 numbness, and 1 hearing disturbance. These episodes preceded a final attack or complete stroke 2 days to 5 months, and those who had a longer period of episodes in the preceding term tended to have less severe deficits. Six of the patients had vertebro-basiler symptoms after being in the upright position, including all the patients in Groups 2 and 4, which had cerebellar border zone/terminal zone infarcts. These results indicate that the hemodynamic mechanism plays an important role in BVAO. The prognosis was not always grave. Four of the patients could walk independently, 2 could walk with a cane, and 3 were bed ridden (2 of which died). Long-term follow-up data (a mean of 5 years) were obtained in all patients. In the patients who could walk, one had asymptomatic cerebellar infarcts, and one had TIAs frequently. Patients with BVAO often also have TIAs and/or preceding episode and show cerebellar border zone/terminal zone infarcts. This research strongly suggests that hemodynamic mechanism might play an important role in BVAO, and that paying attention to border zone infarction in MRI and transient episodes can lead to earlier diagnosis and treatment. PMID- 9396357 TI - [A pedigree of autosomal dominant limb-girdle myopathy with rimmed vacuole formation]. AB - We describe a kindred with autosomal dominant myopathy with preferential proximal limb muscle involvement. This disorder is characterized clinically by early adult onset, slow progression, normal life expectancy, weakness and atrophy of proximal limb muscles, especially in the lower limbs. Laboratory examinations showed myopathic changes mixed with neuropathic components on needle electromyography, slight elevation of serum creatine kinase, and absent cardiac involvement. In biopsied muscle findings of two patients, the presence of rimmed vacuoles was the most striking finding to explain muscle degeneration, though a few necrotic fibers were present. The pathologic and clinical findings in the present family are almost similar to those seen in "adult-onset autosomal dominant limb-girdle muscular dystrophy" reported by Chutkow et al. PMID- 9396358 TI - [Body fat loss in patients with Parkinson's disease]. AB - In order to investigate emaciation in patients with Parkinson's disease (PD), an anthropometric study was undertaken. In 59 out-patients (29 males, 30 females) with PD, the changes in body weight (delta BW) and in body mass index (delta BMI) were obtained and the ratio of body fat (% Fat) was measured with a bioelectrical impedance method. Twenty-two percent of the patients with PD had lost more than 10 Kg of BW (7% of males, 37% of females) and delta BW was -3.9 +/- 7.1 Kg, delta BMI -1.8 +/- 3.2 Kg/m2, and %Fat 25.9 +/- 5.6%. The reductions in both the BW and BMI of females with PD were significantly higher than for males (p < 0.003, p < 0.001, respectively). The disease duration had no correlation with delta BW or delta BMI, but a significant negative correlation with %Fat (all: p < 0.05; females: p < 0.03). However, no correlation was found between delta BW, delta BMI or %Fat on the one hand, and the disease severity or the total dosage of L-dopa with carbidopa on the other. We conclude that patients with PD, especially in females, experience gradual body fat loss according to disease duration. PMID- 9396359 TI - [Immunohistochemical localization of chymase; a mast cell marker and clinical significance in diseased human skeletal muscle]. AB - In the advanced stage of dystrophinopathy, cardiac dysfunction is a serious complication for prognosis. Recently, an angiotensin converting enzyme (ACE), which converts angiotensin (A) 1 to A 2, has been reported to be effective for cardiac insufficiency. The A 2 is produced more dominantly in the path via the production of a neutral serine protease, chymase (MW 25,000), secreted from the mast cell. We have observed localization of chymase in diseased human skeletal muscle tissues, and evaluated its clinical significance. The frozen muscle biopsied specimens from 91 neuromuscular disorders (muscular dystrophies, inflammatory myopathies and neurogenic muscular disorders) were stained by using monoclonal antibody against the chymase, and the positive cells in a whole sectional field were counted. In the serial sections, we also performed routine histochemistry and immunostainings of immunological markers (CD4, CD8 and others) as well as the apoptotic proteins for comparison. RESULTS: The chymase-positive mast cells were scattered mainly in the endomysium, partly in the perimysium and around small vessels. Although the positivity was not disease specific, more numerous strongly positive cells were observed in dystrophinopathy and inflammatory myopathies, but less in myotonic dystrophy and neurogenic muscle disorders. In the normal control muscle, however, strongly positive cells appeared less frequently than in the above mentioned diseased muscles. The chymase-positive cells partly corresponded to the ubiquitin-positive ones, but perforin, granzyme A, Fas and Bcl-2 did not. In conclusion, the chymase-positive mast cell may play a primary or secondary role in the diseased muscle, and their more abundant appearance in dystrophinopathy and some other myopathies suggest the effectiveness of an ACE blocker, an anti-chymase drug. PMID- 9396360 TI - [Two cases of encephalo-myelo-radiculoneuropathy, triggered by herpes simplex virus type-1 infection]. AB - We report two cases of encephalo-myelo-radiculoneuropathy, triggered by herpes simplex virus type-1 (HSV-1) infection. Patient 1 (a 25-year-old man) and patient 2 (a 52-year-old man) were admitted to the hospital because of fever, headache, abnormal behavior, and loss of consciousness. In each case, cerebrospinal fluid (CSF) showed lymphocytic pleocytosis with protein elevation, and serum and CSF IgG antibody titers to HSV-1 were elevated markedly. Although patient 1 was treated with aciclovir in the early phase of encephalitis, he developed severe quadriparesis as a sequela. Patient 2 was treated with a combination of aciclovir and corticosteroids, and he recovered completely about 4 months after the onset of the disease. There have been only a few reports of encephalo-myelo radiculoneuropathy triggered by HSV-1 infection. Early corticosteroid therapy was effective in our patients with post-HSV-1 infectious encephalo-myelo radiculoneuropathy. These two patients were studied with flow cytometry for peripheral blood lymphocyte subsets during the disease course. In the active stage of the disease, the helper-inducer (CD4 + CD29+), activated T cell (CD4 + CD25+), and cytotoxic/NK (CD8 Dull + CD11b Bright+) subsets were increased compared with subsets in controls. An interesting finding was mismatched responses with an increased suppressor-inducer (CD4 + Leu8+) subset and a decreased suppressor-effecter (CD8 Bright+ CD11b Dull+) subset, indicating a possible autoimmune character of encephalo-myelo-radiculoneuropathy triggered by viral infection. PMID- 9396361 TI - [A case of optic-spinal form of multiple sclerosis with lobar type large cerebral hemorrhage]. AB - We report a 57-year-old woman with optic-spinal form of active multiple sclerosis, who developed a large lobar type hemorrhage of the brain. She initially suffered from left visual loss, and three month later, she was hospitalized with paraplegia and total sensory loss up to the fourth thoracic level accompanied by sphincteric disturbance. Diagnosis of clinically probable multiple sclerosis was based on the relapsing-remitting clinical course and laboratory findings. Five months after admission, she developed sudden consciousness loss. Brain CT scan showed a massive hemorrhage in the right frontal to parietal lobe. The patient had no risk factors for cerebral hemorrhage including hypertension. Histopathological study of brain tissues obtained at surgical evacuation of hematoma did not reveal any malignancy, and congo-red staining of this specimen was negative. We analyzed coagulation, fibrinolytic, and endothelial parameters during the follow-up period. von Willebrand factor (vWF) as a marker for endothelial damages was elevated persistently. Moreover, thrombin-antithrombin III complex (TAT) as a marker for activation of coagulation was also elevated constantly throughout the clinical course. The findings suggest that fragility of the vascular walls and permeability changes associated with immunological mechanisms might have resulted in the cerebral hemorrhage. Although there are few reports of cerebral hemorrhage in patients with multiple sclerosis, it has been reported that vascular wall damage is an important aspect of the pathology of multiple sclerosis and acute cerebral vascular damage may sometimes occur in multiple sclerosis. We propose that coagulation studies including the endothelial marker such as vWF would provide a useful information regarding the risk of cerebrovascular complication in multiple sclerosis. PMID- 9396362 TI - [Metamorphopsia and transient increase in the cerebral blood flow of the left occipital pole on 123I-IMP SPECT: a case report]. AB - A 55-year-old right-handed man suddenly developed unformed visual hallucination of rainbow-colored balls coming out from the lower quadrant of the right visual field. Visual field examination revealed a right lower quadrant homonymous hemianopia. Metamorphopsia of the hand or face appeared 6 days later when he looked at his hands or at the face in the mirror, and persisted for about 10 minutes. 123I-IMP SPECT demonstrated a marked increase in CBF of the left occipital pole while the patient realized the visual symptoms, and a marked decrease in CBF after the symptoms disappeared. T1 and T2-weighted MRIs of the brain were unremarkable, but the Gd-DTPA-enhanced T1-weighted MRI showed high signal in the subcortical white matter of the left occipital pole. The metamorphopsia was induced probably by the activation of the left occipital lesion by the epileptogenic mechanism although the nature of the lesion remained unclarified. PMID- 9396363 TI - [A familial case of postural reflex disorder presenting high intensity area mainly in the globus pallidus on T1-weighted cerebral MRI without clear liver damage]. AB - A 56-year-old woman was admitted because of chronic postural reflex disorder. A cerebral MRI revealed symmetrical high intensity area mainly in the globus pallidus on T1-weighted image. The symptom became manifested as gait disturbance from the age of 2 and gradually progressed. Her condition has, however, remained stable since the age of 26. The only sign of parkinsonism was akinesia. There was clear retropulsion but cerebellar ataxia was minimal, and dystonia was negligible. She had no dementia. Her parents were cousins and similar symptoms and high intensity area were found in one of her sisters. Routine liver function tests were normal, with only ICG elevated. Serum copper and ceruloplasmin were normal. A hereditary factor was suspected. There are no similar cases reported in the literature, thus we thought it worth reporting. PMID- 9396364 TI - [A case of antiphospholipid syndrome associated with myasthenia gravis]. AB - We report a 40-year-old Japanese woman with antiphospholipid antibody syndrome (APS) associated with myasthenia gravis (MG). She had a history of miscarriage at the age of 27 followed by pulmonary embolism 3 weeks later. At the age of 40, she developed diplopia, bilateral ptosis and easy fatigability. Serum anti acetylcholine receptor antibody and tensilon test were positive. She was diagnosed as having MG. The laboratory test revealed mild thrombocytopenia, prolonged activated partial thromboplastin time (aPTT) and positive findings for both beta 2-glycoprotein I-dependent anticardiolipin antibody and lupus anticoagulant. She fulfilled the diagnostic criteria of APS, but did not the criteria proposed by American Rheumatism Association for SLE. An extended total thymectomy was performed after administration of oral prednisolone and low-dose aspirin. This is a patient who had APS associated with MGs: both are known to result from autoimmune abnormality. The clinical and laboratory manifestations of APS were ameliorated after removal of the thymus, suggesting that thymectomy alleviates APS symptoms. PMID- 9396365 TI - [A case of idiopathic orthostatic hypotension with selective involvement of postganglionic noradrenergic neurons]. AB - A 44-year-old man had a 30-year history of orthostatic hypotension and diarrhea. One month before admission, he suddenly lost consciousness by defecation, and was hospitalized. He became alert within two days, but he could not sit up due to severe orthostatic hypotension. At that point, he was transferred to our hospital. On admission, talipes and microdactylia were noted. Neurological examination revealed brisk patellar tendon reflexes. Anhidrosis or impotence was not present. Analyses of blood and urine yielded normal results. Cardiological examination revealed no abnormality that could be responsible for the hypotension. MR images of the brain were also normal. However, single photon emission tomography revealed diffuse hypoperfusion of the brain. A head-up tilt (50 degrees) test induced a remarkable fall in systolic blood pressure from 149 (heart rate; 65/min) to 64 mmHg (83). Immersion of hand in ice-cold water failed to increase blood pressure. Heart rate variation and cystometry results, which represent the function of parasympathetic nerves, were normal. Warming of his body caused normal sweating. Intravenous injection of low doses of norepinephrine and methoxamine increased blood pressure while isoproterenol remarkably increased heart rate, suggesting that both alpha- and beta-receptors developed supersensitivity. Instillation of 5% cocaine and 5% tyramine into the conjunctival sac failed to cause pupillary dilation. Clinical findings and pharmacological challenge test results suggested that the main lesion of his autonomic nervous system was selectively confined to the postganglionic sympathetic nerves and noradrenergic (not cholinergic) neurons. The autonomic failure of our patient can be classified as idiopathic orthostatic hypotension. However, most patients with idiopathic orthostatic hypotension or pure autonomic failure complain of anhidrosis and impotence, which were not noted in our patient. These symptomatic differences may be the result of the highly selective involvement of noradrenergic neurons in the postganglionic sympathetic system in our patient. PMID- 9396366 TI - [A case of paradoxical cerebral embolism with 'spectacular shrinking deficit']. AB - A 66-year-old woman was admitted to our hospital because of abrupt onset of left hemiparesis and confusional state. Ten hours prior to admission, she developed difficulty in left hand movement, and her family noticed cloudiness of consciousness and gait disturbance. Neurological examination revealed only a hollow hand sign of the left hand without other neurological deficits. Cranial CT showed a low density area in the right superior parietal region, and the follow up CT on 6th hospital day exhibited contrast enhancement and hemorrhagic transformation in this area. Cerebral angiography demonstrated capillary blush and early venous filling in the right posterior parietal region. Transesophageal color-coded Doppler echocardiography (TEE-CD) showed a shunt flow via patent foramen ovale (PFO). Thus, we diagnosed this case as paradoxical cerebral embolism through PFO, which is characterized by rapid recovery of clinical manifestation, so-called spectacular shrinking deficit. Even in elderly patients with embolic stroke of unknown origin, TEE-CD should be performed to investigate PFO. PMID- 9396367 TI - [A case of adult meningitis with bilateral sensorineural hearing loss at the onset]. AB - Here we report a case of pneumococcal meningitis with bilateral sensorineural hearing loss at the onset. The patient was a 60-year-old man who a few days before visiting our hospital experienced common cold-like symptoms, and then he suddenly developed bilateral hearing loss. Examination of the cerebrospinal fluid (CSF) on the day of admission revealed pleocytosis and his CSF culture demonstrated pneumococci. Otorhinolaryngological examinations disclosed bilateral severe sensorineural hearing loss due to cochlear impairment. Many cases of bacterial meningitis concomitant with hearing loss have been reported, but a case of meningitis starting with sudden hearing loss is rare. PMID- 9396368 TI - [Cerebrospinal fluid neuron-specific enolase as a useful indicator for the early diagnosis of Creutzfeldt-Jakob disease]. PMID- 9396369 TI - [The future of RA treatment]. PMID- 9396370 TI - [Effects of low dose methotrexate therapy in rheumatoid arthritis: a comparison of three different dosage regimens]. AB - The effectiveness of methotrexate (MTX) in treatment of rheumatoid arthritis (RA) was evaluated by following the course of 232 cases. The number of cases in which MTX treatment was assessed as effective amounted to 149 (64.2%), and among these the number of cases in which the effect was excellent amounted to 59 (25.4%). As for the time required to achieve the therapeutic effect, effectiveness was evident within one month in most cases and the more prominent was the result, the shorter was the time required to achieve the effect. In analysis of it's effectiveness, no relationship to the patient's background such as morbidity period, the extent of inflammation, or the period of administration of anti rheumatic drugs was observed and this drug seems promising for treatment of severe inflammation in cases of RA with prolonged course enabling application to a wide range of patients. Also, therapeutic effectiveness was observed in a group of elderly patients, who were more than 65 years old. However, in such cases, MTX should be administered carefully because of its reported strong adverse effects. Its effectiveness and adverse effects have been shown to be dose-dependent. Therefore, as the optimum dosage of MTX, it is recommended to administer a dose of 5.0 mg/week initially for an evaluation period of two months and in those cases in which no effect is observed, the dose is then increased to 7.5 mg/week. PMID- 9396371 TI - [Expression of matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinases (TIMPs) in joint tissues of rapidly destructive coxarthropathy (RDC), analyzed by immunohistochemical study]. AB - OBJECTIVE: Rapidly destructive coxarthropathy (RDC) is characterized by rapid destruction of hip joints, but its pathogenetic mechanism is still obscure. Matrix metalloproteinases (MMPs) are possibly one of the candidates concerning with this mechanism. We attempted histochemical investigation to demonstrate MMPs and tissue inhibitor of metalloproteinases (TIMPs) in joint tissues obtained from RDC patients to clarify their roles in the destruction mechanism. MATERIALS AND METHODS: Joint tissues including synovia and cartilage-bone tissues were obtained from RDC patients at total hip replacement (THR). After fixation with 4% paraformaldehyde, cartilage-bone tissues were partly decalcified. We performed histochemical study for paraffin sections of these tissues by using avidin-biotin method. Antibodies used in this study were monoclonal antibodies to MMP-1, MMP-2, MMP-7, MMP-8, MMP-9, TIMP-1, TIMP-2 and polyclonal antibody to MMP-3. RESULTS: Histological feature of RDC was severe destruction of cartilage and bone by invasion of non-specific granulation tissues composed of many small vessels, macrophages and fibroblastic cells. At the same time, RDC showed apparently fewer lymphocytic cells in these granulation tissues compare with rheumatoid arthritis. MMP-2 and MMP-9 were expressed most demonstrably in synovia and destructive regions of femoral heads, especially in osteoclasts, macrophages, and fibroblastic cells, while MMP-1, MMP-3, were slightly expressed only in the superficial layer of synovia in limited cases. MMP-8, usually contained in neutrophils, was not present in RDC. On the other hand TIMP-1 and TIMP-2 were presented throughout the synovia and destructive regions of femoral heads including fibroblastic cells, macrophages, osteoblasts and osteocytes. CONCLUSION: Immunohistochemical study revealed obvious presence of MMP-2 and MMP 9 in synovia and destructive regions of femoral heads in RDC. Those evidence suggest that MMP-2 and MMP-9 share very important role in the destructive mechanism of RDC, possibly under imbalance between TIMPs. PMID- 9396372 TI - [A case of polymyalgia rheumatica with excessive increase of rheumatoid factor]. AB - A 76 year-old woman suffered from muscle pain and stiffness of acute onset in her shoulder girdle and pelvic girdle, which were followed by mild left temporal headache and transient arthralgia. Neither joint swelling nor sicca symptom was observed. Laboratory data showed high ESR (128 mm/hr), positive CRP (12.9 mg/dl), increased fibrinogen (485 mg/dl) and high titer of rheumatoid factor (RF) (RAHA x 640). Other autoantibodies examined were negative. Muscle enzymes and electromyogram were within normal limits. Joint X ray didn't reveal the finding suggestive of RA. After the treatment with prednisolone (PSL) 15 mg/day, clinical symptoms and laboratory data improved dramatically. Though she had excessive increase of RF (RAHA x 10240) during therapy, no recurrence of articular symptoms were recognized. She continues to be well with PSL 5 mg/day after 1 year 5 months from onset. As for polymyalgia rheumatica (PMR) followed by RA, the appearance or exacerbation of arthritis corresponding to the elevation of RF occurred in all previously reported 17 cases. Recurrence of arthralgia corresponding to the elevation of RF was not recognized in this case. In addition, Hunder et al reported that PMR with little or no observable joint swelling after several weeks of symptoms is unlikely to develope RA. Therefore, it is speculated that this case in unlikely to develope RA and assessment of arthritis corresponding to the elevation of RF is important to differentiate PMR and elderly-onset RA. This case of PMR is the 5th case with excessive increase of RF in Japan. PMID- 9396373 TI - [Two cases with SLE and MCTD developed after a long period of chronic arthritis that was initially diagnosed as JRA]. AB - In order to discuss the diversity of clinical features and the difficulty in diagnosis of children with juvenile rheumatoid arthritis (JRA), we present two cases who have documented the development of systemic lupus erythematosus (SLE) and mixed connective tissue disease (MCTD) after a long period of disease characterized only by arthritis that was initially diagnosed as JRA. The first case was a girl diagnosed for her arthritic joints as polyarticular JRA at 15 years of age. At onset, she had Raynaud phenomenon but autoantibodies such as anti-nuclear antibody (ANA), anti-DNA antibody, and rheumatoid factor were negative. Five years after onset, she became ANA positive and 3 years later she became pregnant. During her pregnancy, she became positive for anti-DNA antibody without any signs of nephritis. One month after the delivery, however, she developed butterfly rash, carditis, nephritis, and was diagnosed as SLE. No destructive changes were observed in her joints though arthritis continued for 8 years form onset to pregnancy. The second case was a 3 years old girl who was diagnosed as polyarticular JRA. Treatment by aspirin induced complate remission after one year from the onset. However, 10 years after that remission, she developed Raynaud phenomenon and arthralgia in her knees and hip joints. Her laboratory findings showed hypergammaglobulinemia, positive ANA, positive anti DNA antibody, positive anti-RNP antibody. She was eventually diagnosed as MCTD when she was found to have polymyositis by EMG and serum CK. In the present paper, two cases imply the difficulty in diagnosing JRA and diversity of rheumatic diseases such as JRA, SLE and MCTD. Closer and longer period of observation is essential for the JRA patients with nondestructive arthritis. PMID- 9396374 TI - [A case of lupus myocarditis and nephritis with transient foramen jugular syndrome]. AB - A 46-year-old man was admitted to our clinic because of acute heart failure. Six years before admission he was pointed out cardiomegary and hematuria. One year later, he was diagnosed as having jugular foramen syndrome. On admission, he had a fever and dyspnea. Pansystolic blowing murmur was audible at the apex. The chest ratio on his chest X-ray was 52.5%. An electrocardiogram showed left ventricular hypertrophy. An echocardiogram showed marked dilatation and severe dysfunction of left ventricle. Radionuclide scanning with technetium 99 m pyrophosphate identified inflammatory change in the apex. Myocardial biopsy showed fibrotic degeneration and IgG deposits in myocardium. Blood examination showed anemia, lymphopenia. positive anti-nuclear antibody (1000 times, shaggy pattern), positive anti ds-DNA antibody and hypocomplementemia. Furthermore, proteinuria was pointed out. Renal biopsy showed focal segmental glomerulonephritis with active necrotizing lesion (type III nephritis). Lupus myocarditis and nephritis was diagnosed. After prednisolone (80 mg/day) was administered. left ventricular function and hypocomplementemia improved. The ACE inhibitor was also used for proteinuria. In spite of a little amount of blood transfusion, he showed hepatic hemosiderosis. We suspect that the cause of hemosiderosis was related chronic inflammation of active lupus. It was treated with Erythropoietin. PMID- 9396376 TI - [Successful treatment of interstitial pneumonia with lipo-PGE1 and pentoxifylline in a patient with dermatomyositis]. AB - Interstitial pneumonia complicated with dermatomyositis sometimes shows a resistance to high dose steroid therapy and a fatal course particularly in patients without showing the elevation of creatine kinase. We experienced a 48 year old woman who developed heliotrope rash, Gottron's sign, multiple cutaneous ulcers, and dyspnea on exertion. These symptoms were resistant to low dose steroid therapy. Serum levels of creatine kinase were normal. Anti-nuclear antibodies and anti-Jo-1 antibody were negative. High resolution CT scan of the chest showed areas of multiple air space consolidation and subpleural linear shadows. Lung biopsy performed under video-assist thoracosurgery revealed diffuse alveolitis with scattered lymphoid folicules and mild accumulation of macrophages in the alveolar spaces. There were no honey-combing. These features were compatible with "non-specific interstitial penumonia" proposed by Katzenstein, 1995. The patient was treated with 10 micrograms lipo-PGE1, PGE1 incorporated in lipid microspheres, and 300 mg pentoxifylline, which resulted in a dramatic improvement of both interstitial pneumonia and cutaneous ulcers. The present case suggested a novel strategy for the treatment of interstitial pneumonia. PMID- 9396375 TI - [A case of amyopathic dermatomyositis associated with interstitial pneumonitis]. AB - A patient (47-year old female) who had erythema similar to Gottron's sign on bilateral finger joints since two years ago, started to have polyarthralgia on bilateral knee and shoulder on the spring of 1993. Polyarthralgia was extended to both wrist and hand joints on Oct. of 1995. On the middle of Dec. 1995, she began to have exertional dyspnea and was referred and admitted in our hospital on 18th, Dec., 1995. Chest X-ray and CT scan showed the shadow for active interstitial pneumonitis on bilateral lower lung fields. Blood gas analysis indicated hypoxia (PaO2: 62.8 mmHg) and low % DLCO (64.7%). Skin eruption of face (heliotrope-like erythema) and hands (Gottron's sign) and skin biopsy (right hand) findings were compatible with that in dermatomyositis. The analysis in blood biochemistry showed no elevation for muscle enzymes. The diagnosis for amyopathic dermatomyositis (ADM) was made according to the criteria proposed by Euwer & Sontheimer (1993). The steroid pulse therapy and 50 mg per day of cyclophosphamide were immediately administered. The dyspnea and dermatitis were improved within two weeks after therapy. She is presently in remission until Jan. 1997 with the maintenance dosis of 10 mg per day of oral prednisolone. PMID- 9396377 TI - [The organ involvement and treatment in SLE]. PMID- 9396378 TI - [Cellular mechanisms of bone resorption in rheumatoid arthritis]. PMID- 9396379 TI - Particulate air pollution and daily mortality: can results be generalized to Latin American countries? AB - OBJECTIVE: Recently, a series of reports, based on ecological analyses of routinely collected data, have shown positive associations between measures of particle concentration and daily mortality counts in various cities of the US and Europe. MATERIAL AND METHODS: We reviewed the process of generalization of these results to Latin American countries addressing possible differences in air pollution mixtures, exposure profiles, and population susceptibility. RESULTS: A limitation to the process of generalization is the lack of a well-established biological mechanism by which particles may act on daily mortality. Also, sources and levels of ambient air pollution as well as population characteristics and habits vary widely between Northern communities of Europe and the US, and Latin American countries, which impairs the process of generalization. However, results of studies conducted in Latin American countries suggest a similar effect to that observed in Northern countries of Europe and the US. CONCLUSIONS: Despite uncertainty about the mechanism, there is sufficient evidence that particles are harmful for health. Control measures of particle emission are urgently needed in Latin American countries. Given the potential of misclassification of exposure, the dose-response relationship observed in Northern Europe and the US may not be adequate for Latin American populations. There is a need for a new generation of epidemiological studies including a specific assessment of exposure to fine particles and of events surrounding death. PMID- 9396380 TI - The challenge and prize for international health organizations in the Americas. PMID- 9396381 TI - The Soteria-concept. Theoretical bases and practical 13-year-experience with a milieu-therapeutic approach of acute schizophrenia. PMID- 9396382 TI - [The delusion of theft in elderly with dementia--(2) Pathogenesis of the delusion; dynamic and structural aspects]. PMID- 9396383 TI - [Clinical examination on localization-related epilepsies with elementary visual seizures--clinical, electroencephalographic and imaging diagnostic studies]. AB - The clinical, electroencephalographic and Imaging diagnostic features of the 45 patients of Localization-related Epilepsy who had elementary visual symptoms at seizure onset were investigated. There were 24 males and 21 females aged 19 to 78. Their ages at seizure onset ranged from 1 to 55 with a mean of 15.4. The patients were divided into 3 groups based on the pattern of elementary visual symptoms: Group 1; 23 patients (51%) with only elementary positive visual symptoms, Group 2; 9 patients (20%) with only elementary negative visual symptoms, Group 3; 13 patients (29%) with others. It came to our notice that 8 (18%) patients in Group 3 had positive + negative (P*N) visual seizures such as scintillation scotoma. It was quite difficult to draw a distinction between the P*N seizures and scintillation scotoma in migrainous patients in quality. Therefore, a question arises whether similarities between them are ascribed to the same underlying mechanism. It is possible that the mechanism of the P*N seizures is different from that of the scintillation scotoma, but the both produce the same condition. However, we are not competent to discuss this hypothesis. Further work along this line is necessary. In addition to the elementary visual symptoms, autonomic (69%), focal motor (29%), illusion (29%), vertiginous (22%) manifestations, etc. were also observed. These manifestations suggest that epileptic ictal discharges spread into many different brain areas. Regarding Group 1-3, occurrence of illusional seizures was more common in Group 2. In this group, in only one patient, occipital interictal discharges were observed. The subjects were subclassified into two groups depending on whether the seizures were well controlled (good outcome) or poorly controlled (poor outcome group). On these groups, comparative studies were performed. The poportion of the patients with a family history of convulsive disorder was higher in the latter than that in the former. To the contrary, the poportion of the patients with a etiologic episode was higher in the former than that in the latter. With respect to the imaging study, regardless of localizations, abnormal findings were detected in 25% on CT scan, in 29% on MRI, and in 71% on SPECT. Whether or not these abnormal findings completely or partially agreed with presumed epileptogenic brain region (occipital lobes) was investigated. CT scan was positive (correspondent) in 8%, MRI was positive in 13%, and SPECT was positive in 54%. The incidence of SPECT abnormalities was higher in the poor outcome group (70 %) than that in the good outcome one (43%). We may, therefore, conclude that SPECT seems to be useful for the detection of epileptogenic region, especially in intractable cases. But the incidence of these imaging studies' abnormalities located in occipital lobe was not so high, therefore, there seems to be no doubt that clinical symptoms and EEG findings are most important for diagnosis. PMID- 9396384 TI - Synthesis and pharmacological activity of two derivatives of the amide of valproic acid. AB - Valproic acid (VPA), a synthetic branched-chain fatty acid, and its pro-drug the primary amide (VPD) are effective and widely used anti-epileptic agents. Although the use of VPA has grown in recent years, major side effects are still associated with this drug. We presume that it is possible, without loosing the VPD pharmacological profile, to obtain new compounds by undertaking substitutions in the CONH group. N,N'-bis-(2-propylpentanoyl)- 1,2-ethanediamine (3) and N,N'-bis (2-propylpentanoyl)-1,3-propanediamine (4) were obtained from VPA (1) using a method reported in the literature. The chemical structures of the new compounds were demonstrated by elemental analysis, IR, and 1H NMR spectroscopy. Both compounds are less toxic and more effective in protecting the animals from death caused by PTZ than VPD after intraperitoneal administration to mice. PMID- 9396385 TI - Synthesis and antitumor activity of 9-anilino, phenylhydrazino, and sulphonamido analogs of 2- or 4-methoxy-6-nitroacridines. AB - Synthesis of several new 9-anilino, phenylhydrazino, and sulphonamido analogs of 2- or 4-methoxy-6-nitroacridine derivatives is described. The prepared compounds were tested for their in vitro antitumor activity against 60 human tumor cell lines by the National Cancer Institute (NCI) and showed a potential anticancer activity. Compounds 9-(phenylhydrazino)-2-methoxy-6-nitroacridine (8a) and 9-(4 chlorophenylhydrazino)-4-methoxy-6-nitroacridine (9b) exhibited a broad spectrum antitumor activity with full panel (MG-MID) median growth inhibition (GI50), of 16.1 and 10.9 microM and total growth inhibition (TGI) of 66.7 and 37.9 microM, respectively. Meanwhile, compounds 15a and 15b showed moderate selectivity toward leukemia cell lines. As a trial to explore the mode of action of their antitumor activity, the 6-nitroacridine analogs were evaluated for their inhibitory effect on major cell cycle control proteins cdc2 kinase and cdc25 phosphatase as possible molecular targets that may account for antimitotic potency. None of the tested compounds proved to exert their activity via this antimitotic mode of action. PMID- 9396386 TI - Studies in sugar chemistry. VII. Glucuronides of podophyllum derivatives. AB - The antitumor activities of several glucuronide methyl esters of podophyllum derivatives were tested in vitro against two human tumor cell lines and their drug resistant sublines. The most active compound studied was methyl (4' carbobenzoxy-4'-demethyl-epipodophyllotoxin-D-glucopyranoside)uronat e 19. Compound 19 was as potent in a colon carcinoma model and was twice as potent in a lung carcinoma model as etoposide 6. In vivo, however, in a mouse leukemia P388 model, it had only marginal activity, with a maximum T/C% value of 125 at 37 mg/kg (iv). PMID- 9396387 TI - Synthesis and calcium channel modulating effects of isopropyl 1,4-dihydro-2,6 dimethyl-3-nitro-4-(thienyl)-5-pyridinecarboxylates. AB - A group of racemic isopropyl 1,4-dihydro-2,6-dimethyl-3-nitro-4-(thienyl)-5 pyridinecarboxylates++ + 7a-f were prepared using a modified Hantzsch reaction that involved the condensation of a thienylcarboxaldehyde 4a-f with isopropyl 3 aminocrotonate 5 and nitroacetone 6. In vitro calcium channel antagonist activities were determined using a guinea pig ileum longitudinal smooth muscle (GPILSM) assay. Compounds 7a-f exhibited weaker calcium channel antagonist activity (IC50 = 10(-5) to 10(-7) M range) than the reference drug nifedipine (IC50 = 1.43 x 10(-8) M). The point of attachment of the C-4 thienyl ring system was a determinant of antagonist activity [3-thienyl (7b) > 2-thienyl (7a)]. A 5 substituent in the 2-thienyl moiety influenced antagonist activity where the potency order was 5-bromo-2-thienyl 7f > or = 5-methyl-2-thienyl 7c > 2-thienyl 7a. Although the 5-methyl-2-thienyl 7c and 3-methyl-2-thienyl 7d isomers are equipotent antagonists, the 5-bromo-2-thienyl compound 7f appears to be marginally more active than the 4-bromo-2-thienyl isomer 7e. The 2-thienyl compound 7a, unlike the 3-thienyl isomer 7b, exhibited an agonist effect on GPILSM in the absence of the muscarinic agonist carbachol. Effects of the 2 thienyl 7a and 3-thienyl 7b isomers on the magnitude of calcium current were determined in guinea pig ventricular myocytes with voltage clamp techniques. Results showed that 2-thienyl 7a inhibited calcium current (antagonist) when voltage steps were made from a potential of -40 mV. However, when voltage steps were made from -60 mV, 7a enhanced calcium current (agonist). The 3-thienyl isomer 7b had little, if any, effect on calcium current. PMID- 9396388 TI - Synthesis, uterotrophic, and antiuterotrophic activities of some estradiol derivatives containing thiadiazole, thiazoline, and thiazolidinone moieties. AB - The effect of structural modification on the biological activity of hormones has been studied on five novel series of estradiol analogs bearing a variety of substituents at the 2-position of the steroidal nucleus. The synthesized compounds include 2-[2-(5-substituted amino-1,3,4-thiadiazol-2-yl)vinyl]estradiol 17 beta-acetate 5-9, 2-aroylmethylestradiols 10-12, 2-[2-aryl-2-(substituted thiocarbamoylhydrazono)ethyl]estradiols 13-18 and their cyclic thiazoline 19-24, and thiazolidinone derivatives 25-30. Among the products, the p hydroxybenzolmethylestradiol 12 exhibited the highest antiestrogenic activity of 63%. It also elicited 34% of the uterotrophic activity of estradiol. PMID- 9396389 TI - Microsomal catalyzed N-hydroxylation of guanfacine and reduction of N hydroxyguanfacine. AB - Guanfacine 1 is a well known centrally acting alpha 2-adrenoceptor agonist with antihypertensive activity. The drug belongs to the guanidine class of compounds. The N-oxidative biotransformation of guanfacine was investigated in vitro in the presence of hepatic microsomal fractions from rabbits, pigs, and humans. The N hydroxylated derivative N-hydroxyguanfacine 2 had to be synthesized as reference for the identification of the metabolite. The corresponding in vitro retroreduction of N-hydroxyguanfacine 2 was also investigated using the same set of enzyme sources. A new HPLC analytical method was developed in order to isolate and quantify the metabolites. A complete metabolic cycle involving the oxidative metabolism of guanfacine could only be observed in the presence of microsomal fractions from rabbitt livers, whereas enzyme sources of all species under investigation catalyzed the N-reduction of N-hydroxyguanfacine 2. PMID- 9396390 TI - Synthesis and evaluation of a novel series of pyrrolizine derivatives as dual cyclooxygenase-1 and 5-lipoxygenase inhibitors. AB - The aim of our study was to investigate structure activity relationship following the replacement of the 6-phenyl substituent at the 6,7-diaryl-2,3 dihydropyrrolizine template by various heteroaromatic residues. In this context we developed a new, efficient, and highly sensitive test method for the screening of dual cyclooxygenase-1 (COX-1) and 5-lipoxygenase (5-LOX) inhibitors. We used human platelets as a source of COX-1 and human PMNLs as a source of 5-LOX. Both cell types were isolated from the same volume of blood. PGE2 and LTB4 respectively were determined by highly selective and sensitive ELISA kits, using monoclonal antibodies. For a single determination at most 0.5 mL whole blood is needed. PMID- 9396391 TI - An improved synthesis of the anti-picornavirus flavone 3-O-methylquercetin. PMID- 9396392 TI - Chemosensitive gliomas in adults: which ones and why? PMID- 9396393 TI - Salvage chemotherapy with paclitaxel for recurrent oligodendrogliomas. AB - PURPOSE: A prospective phase II study of paclitaxel was performed in adult patients with recurrent hemispheric oligodendrogliomas. PATIENTS AND METHODS: Twenty adult patients (14 men and six women), ages 18 to 52 years (median, 40.5), with recurrent supratentorial hemispheric oligodendrogliomas were treated. All patients had previously been treated with surgery, involved-field radiotherapy (median dose, 55 Gy; range 54 to 55 Gy) and nitrosourea-based (procarbazine, lomustine [CCNU], and vincristine [PCV-3 regimen]) chemotherapy (median number of cycles, five; range, four to six). Fourteen patients were treated adjuvantly with radiotherapy and nitrosourea-based chemotherapy; six were treated at recurrence following initial gross total resection with reoperation (subtotal resection in all), radiotherapy, and nitrosourea-based chemotherapy. Paclitaxel was administered intravenously at a dose of 175 mg/m2 every 3 to 4 weeks with neurologic and neuroradiographic evaluation every 8 weeks. RESULTS: A median of three cycles of paclitaxel (range, two to 10) were administered. All patients were assessable. Toxicity included partial alopecia (12 patients), thrombocytopenia (six), neutropenia (three), and anemia (one). One patient developed neutropenic fever without bacteriologic documentation and four required transfusion of blood products (RBCs, n = 2; platelet, n = 2). No treatment related deaths occurred. Ten patients (50%) demonstrated either a neuroradiographic partial response (n = 3) or stable disease (n = 7), with a median response and stable disease duration of 10 months (range, 5 to 14). CONCLUSION: Paclitaxel demonstrated modest efficacy with minimal toxicity in this pretreated cohort of adult patients with recurrent hemispheric oligodendrogliomas. PMID- 9396395 TI - Gemcitabine: a promising new agent in the treatment of advanced urothelial cancer. AB - PURPOSE: To evaluate the efficacy and toxicity of gemcitabine (2',2' difluorodeoxycytidine) in previously untreated patients with advanced transitional cell carcinoma. PATIENTS AND METHODS: Forty-one patients with measurable advanced transitional cell carcinoma who had received no prior chemotherapy for metastatic disease were scheduled to receive gemcitabine 1,200 mg/m2 intravenously over 30 minutes on days 1, 8, and 15 of a 28-day cycle. Prior adjuvant or neoadjuvant therapy for locally advanced disease was allowed if this was completed greater than 1 year prior to study entry. All patients were treated on an outpatient basis. RESULTS: There were three complete responses and six partial responses seen in 37 assessable patients, for an overall response rate of nine of 37 (24.3%; 95% confidence interval, 12 to 41). Four patients remain in remission at 14, 23, 24, and 31 months. The median survival was 8 months with 17% of patients alive at 2 years. Treatment generally was well-tolerated with three patients having > or = grade 3 nonhematologic toxicity, five having grade 3 neutropenia, two having grade 3 thrombocytopenia, and two episodes of febrile neutropenia. Most patients were able to receive the drug as scheduled with the primary reason for dose reduction or dose delay being neutropenia. CONCLUSION: Gemcitabine has promising single-agent activity against urothelial cancer with a favorable toxicity profile. Further studies in combination with other active agents are warranted. PMID- 9396394 TI - NB87 induction protocol for stage 4 neuroblastoma in children over 1 year of age: a report from the French Society of Pediatric Oncology. AB - PURPOSE: NB87 was designed to test the efficacy of a short, non cross-resistant, induction protocol for unselected patients over 1 year of age with stage 4 neuroblastoma. A secondary objective was to compare in a randomized study the toxicity of two modalities of cisplatin administration. PATIENTS AND METHODS: A total of 183 patients received two cycles of alternating sequences: cyclophosphamide 300 mg/m2/d on days 1 to 5, vincristine 1.5 mg/m2/d on days 1 and 5, and doxorubicin 60 mg/m2/d on day 5 (CADO); and cisplatin 40 mg/m2/d and etoposide 100 mg/m2/d on days 1 to 5 (CVP), followed by surgery of the primary tumor (126 patients). Ninety-one were randomized to receive cisplatin either as bolus (BO; n = 48) or continuous infusion (CI; n = 43). International Neuroblastoma Staging System (INSS) and Response Criteria (INRC) were used with emphasis on skeletal evaluation by meta-iodobenzylguanidine (MIBG). RESULTS: Hematotoxicity was predominant, with a higher incidence of neutropenia (P = .01) for CADO and of thrombocytopenia for CVP (P < .001). Severe infections, as well as nonhematologic toxicities, occurred more often after the first sequence. Gastrointestinal complications were predominant during both courses of CVP. The toxic death rate, including surgery, was 3%. Complete remissions (CRs) were less frequent on MIBG (45%) compared with marrow (66%) or other metastases (61%). Combining all metastatic sites resulted in a 39% CR rate. After surgery, the final CR rate was 42%. Nephrotoxicity was minimal in both arms (92% normal clearance for CI v 82% for BO). Hearing loss greater than 40 dB at 6,000 to 8,000 Hz was reported equally in both arms (n = 6 for CI v n = 5 for BO). CONCLUSION: Intensified chemotherapy using CADO/CVP increases CR rates despite a shorter induction duration. However, the rate of MIBG normalization remains unsatisfactory and could be raised through the dose-intensive use of agents such as cyclophosphamide. PMID- 9396396 TI - Serum tissue polypeptide antigen in bladder cancer as a tumor marker: a prospective study. AB - PURPOSE: Tissue polypeptide antigen (TPA) is a differentiation and proliferation tissue marker of epithelia. Increased serum levels were found in some patients with invasive bladder cancer. We present the results of a prospective study that evaluated the role of serum TPA (S-TPA) in bladder carcinoma. PATIENTS AND METHODS: The series included 167 patients treated for invasive bladder cancer between 1989 and 1996. S-TPA concentrations were measured by radioimmunoassay before treatment, at the end of treatment, and during follow-up evaluation. The upper normal limit of the test was set at 80 UI/L. RESULTS: With a specificity of 100%, the diagnostic sensitivity was 46%. Pretherapeutic S-TPA levels were significantly correlated with tumor stage (T2 v T3 and T4; P = .02), with nodal stage (N0 v N1 and N2; P = .00001), and with metastatic stage (M0 v M1; P = 10[ 6]), but not with histologic grading (grade 1 and 2 v 3). In the subset of patients with normal pretherapeutic S-TPA levels, 95% had no residual disease at the end of treatment, compared with 53% of patients with initial elevated S-TPA (P = 10[-8]). Among patients who achieved a complete response, 27% experienced a relapse, with an increase of S-TPA in 72% of cases. The mean follow-up time was 20 +/- 17 months. For patients with normal pretherapeutic S-TPA levels, 3-year overall survival and disease-free survival rates were 76% and 67% respectively. These were 46% (P = .001) and 25% (P = 10[-7]), respectively, for patients with high pretherapeutic S-TPA. Multivariate analysis showed that S-TPA was an independent prognostic factor for survival (P = .03). CONCLUSION: In invasive bladder cancer, S-TPA level is correlated with initial tumor stage. It is a valuable parameter for follow-up evaluation and appears to be a prognostic factor in multivariate analysis. PMID- 9396397 TI - Presence of circulating prostate cells in the bone marrow of patients undergoing radical prostatectomy is predictive of disease-free survival. AB - PURPOSE: To determine whether the presence of circulating prostate cells in the bone marrow is associated with disease-free survival in patients undergoing radical prostatectomy. MATERIALS AND METHODS: We evaluated the bone marrow of 86 patients with clinically localized prostate cancer treated by radical prostatectomy for the presence of circulating prostate cells using reverse transcriptase polymerase chain reaction (RT-PCR) amplification of prostate specific antigen (PSA) mRNA. Follow-up duration ranged from 1 to 43 months (mean, 15.4). RESULTS: Two of 47 patients (4%) with negative RT-PCR PSA results and 10 of 39 patients (26%) with positive RT-PCR PSA results have had disease recurrence. Patients whose RT-PCR PSA results were positive had a significantly shorter disease-free survival period than those patients with negative RT-PCR PSA results (P = .004). RT-PCR status correlated significantly with serum PSA level (P = .001) and pathologic stage (P = .003). Based on Cox's proportional hazards models, RT-PCR status was found to be a significant predictor of disease-free survival. However, after controlling for PSA level, RT-PCR status was not significant in predicting disease-free survival. CONCLUSION: RT-PCR PSA of bone marrow may be a useful pretreatment prognostic test for patients undergoing radical prostatectomy. Currently, this test should not be used to determine if patients receive definitive local therapy. PMID- 9396398 TI - Retinoblastoma protein and proliferating-cell nuclear antigen expression as predictors of recurrence in well-differentiated papillary thyroid carcinoma. AB - PURPOSE: We analyzed retinoblastoma protein (pRB) and proliferating-cell nuclear antigen (PCNA) expression in primary tumors and recurrent lesions of well differentiated papillary thyroid carcinoma (PTC) to clarify the relationship between their expression and recurrent disease. PATIENTS AND METHODS: The study included 93 patients with PTC. No recurrent disease had developed in 60 patients within 10 years after surgery (group N). Thirty patients in whom recurrent disease had developed after surgery were enrolled in group R. Levels of pRB and PCNA expression were quantified using the CAS 200 system (Cell Analysis Systems, Elmhurst, IL) following immunohistochemical staining. RESULTS: Mean pRB expression level in the primary tumors in group R was significantly lower than that in group N (P < .0001). pRB expression in the tumors with a diameter up to 20 mm was significantly lower than that in tumors larger than 20 mm in group R (P < .01). There were no significant differences in the levels of expression of PCNA in the primary tumors between group N and group R. Univariate analysis demonstrated that the disease-free survival was significantly correlated with pN category, pRB, and PCNA expression level. The subgroup with high-level expression of pRB (> 25%) showed significantly long disease-free survival (P < .001). Furthermore, the subgroup with low-level expression of PCNA (< 35%) showed significantly longer disease-free survival (P < .05). Multivariate analysis showed pRB expression and pN category to be independent prognostic factors for disease-free survival in PTC. CONCLUSION: pRB expression level can be used as a reliable predictor for recurrence of PTC. PMID- 9396400 TI - Chronologic changes in the clinicopathologic findings and survival of gastric cancer patients. AB - PURPOSE: To determine the chronologic changes in the clinicopathologic features of gastric cancer patients. PATIENTS AND METHODS: The clinicopathologic findings of 1,795 patients with gastric cancer were examined retrospectively from hospital records obtained between 1969 and 1995. The patients were divided into three generations on the basis of chronologic order. The first generation included patients treated over the period 1969 to 1977; the second generation, 1978 to 1986; and the third generation, 1987 to 1995. RESULTS: The chronologic changes in the clinicopathologic findings for all gastric cancers included increases in the superficial type based on macroscopic appearance (P < .005), small-sized tumor (P < .025), superficial depth of invasion (P < .005), and earlier histologic stages (P < .005), in addition to a decrease in lymph node metastasis (P < .005). Overall, the postoperative survival rate has improved over time in gastric cancer patients, with 5-year survival rates of 36.0%, 53.3%, and 68.6% in the first, second, and third generations, respectively. In stages 1,2, and 3, the survival rate in the third generation was the highest of the three generations, whereas in stage 4, the survival rate did not differ between the three generations. Patients who underwent a D2 dissection showed a higher survival rate than those with D1 or D3 dissections, but there was no statistical difference in the survival of patients with D1, D2, and D3 dissections when stage 4 patients were excluded. CONCLUSION: The chronologic changes in gastric cancer patients over the past 27 years have included an increase in the incidence of earlier-staged gastric cancers, which has had a significant impact on the improved postoperative survival rate. PMID- 9396399 TI - Paclitaxel, carboplatin, and extended-schedule etoposide in the treatment of small-cell lung cancer: comparison of sequential phase II trials using different dose-intensities. AB - PURPOSE: In two sequential phase II studies, we evaluate the feasibility and efficacy of adding paclitaxel to a standard platinum/etoposide regimen in the first-line treatment of small-cell lung cancer. PATIENTS AND METHODS: One hundred seventeen patients with small-cell lung cancer were treated between June 1993 and July 1996. The first 38 patients received a lower-dose regimen: paclitaxel 135 mg/m2 by 1-hour infusion, carboplatin at an area under the concentration-time curve (AUC) of 5.0, and etoposide 50 mg alternating with 100 mg orally on days 1 to 10. When only mild myelosuppression was observed, doses of paclitaxel and carboplatin were increased in the subsequent 79 patients (paclitaxel 200 mg/m2 by 1-hour infusion and carboplatin at an AUC of 6.0). All patients received four courses of treatment, administered at 21-day intervals. Patients with limited stage small-cell lung cancer also received thoracic radiation therapy (1.8 Gy/d; total dose, 45 Gy) administered concurrently with courses 3 and 4 of chemotherapy. RESULTS: Seventy-two of 79 patients (91%) who receive the higher dose regimen had major responses. Thirty-two of 38 (84%) with extensive-stage disease responded (21% complete response rate); median survival was 10 months for this group. With limited-stage disease, the overall response rate was 98%, with 71% complete responses; the median survival time has not been reached at 16 months. Median survival in extensive-stage patients was longer in patients who received the higher-dose regimen (10 months) than in the previous group treated with lower doses (7 months; P = .008). The higher-dose regimen was well tolerated, with myelosuppression being the major toxicity. Compared with the lower-dose regimen, grade 3/4 neutropenia increased from 8% to 38% of courses, but the incidence of hospitalization for neutropenia and fever did not increase. Other nonhematologic toxicities were uncommon, and did not increase substantially with the higher-dose regimen. CONCLUSION: Paclitaxel can be added at full dose (200 mg/m2) to a carboplatin/etoposide combination while maintaining a tolerable toxicity profile. Median survival times in both extensive- and limited-stage patients compare favorably with other reported regimens. This regimen merits further investigation, and a randomized trial to compare this regimen with a standard carboplatin/etoposide combination is underway. PMID- 9396401 TI - Preoperative chemotherapy for stage IIIB extremity soft tissue sarcoma: long-term results from a single institution. AB - PURPOSE: To review a single institution's long-term results with doxorubicin based preoperative chemotherapy for American Joint Committee on Cancer (AJCC) stage IIIB extremity soft tissue sarcoma (STS). PATIENTS AND METHODS: The records of all patients with AJCC stage IIIB extremity STS treated with preoperative chemotherapy between 1986 and 1990 at The University of Texas M.D. Anderson Cancer Center were reviewed to assess rates of response, local recurrence-free survival (LRFS), distant metastasis-free survival (DMFS), disease-free survival (DFS), and overall survival (OS). RESULTS: Seventy-six patients with stage IIIB disease received preoperative chemotherapy. The median sarcoma size was 10 cm. Seventy-two patients (95%) had tumors located deep to the muscular fascia. Most patients received a median of three preoperative cycles of doxorubicin and dacarbazine (ADIC), cyclophosphamide and ADIC (CyADIC), or other doxorubicin based regimens. Radiographic response rates were as follows: complete response (CR), 9%; partial response (PR), 19%; minor response, 13%; stable disease, 30%; and progression, 30%. The overall objective major response rate (CRs plus PRs) was 27%. At a median follow-up time of 85 months, 5-year actuarial rates of LRFS, DMFS, DFS, and OS with 95% confidence intervals (CIs) were 83% (CI, 73% to 94%), 52% (CI, 41% to 66%), 46% (CI, 35% to 60%), and 59% (CI, 48% to 72%), respectively. Comparison of responding patients (CRs plus PRs) and nonresponding patients did not show any significant differences in LRFS, DMFS, DFS, or OS. CONCLUSION: Preoperative doxorubicin-based chemotherapy was associated with response, DFS, and OS rates similar to those observed in randomized postoperative chemotherapy trials. Responding patients had rates of LRFS, DMFS, DFS, and OS comparable to those of nonresponders. PMID- 9396402 TI - Rapid recovery of spermatogenesis after mitoxantrone, vincristine, vinblastine, and prednisone chemotherapy for Hodgkin's disease. AB - PURPOSE: Because the effects of mitoxantrone on human male fertility were unknown, we determined prospectively the effects of three courses of mitoxantrone (Novantrone), vincristine (Oncovin), vinblastine, prednisone (NOVP) chemotherapy on the potential for fertility of men with Hodgkin's disease (HD). PATIENTS AND METHODS: Semen analyses were performed on 58 patients with stages I-III HD before, during, and after chemotherapy and after the sperm count recovered from the effects of abdominal radiotherapy that was given after chemotherapy. RESULTS: Before the initiation of treatment, 84% of the patients were normospermic. Sperm counts declined significantly within 1 month after the start of NOVP chemotherapy. In the month after chemotherapy, 38% of patients were azoospermic, 52% had counts < 1 million/ mL, and 10% had counts between 1 and 3 million/mL. Between 2.6 and 4.5 months after the completion of chemotherapy, sperm counts recovered rapidly to normospermic levels in 63% of patients. In the remaining patients who were followed up for at least 1 year after standard upper abdominal radiotherapy, counts also recovered to normospermic levels. CONCLUSION: NOVP chemotherapy, like most other regimens, produced marked temporary effects or spermatogenesis. However, sperm production recovered very rapidly, within 3 to 4 months after the end of NOVP chemotherapy. This pattern was caused by killing differentiating spermatogenic cells, but there was little cytotoxicity or inhibition of stem cells from mitoxantrone or the other drugs. After the combination of NOVP plus abdominal radiotherapy, sperm counts and motility were restored in most patients to pretreatment levels, which were compatible with normal fertility. PMID- 9396403 TI - Value of different modalities of granulocyte-macrophage colony-stimulating factor applied during or after induction therapy of acute myeloid leukemia. AB - PURPOSE: The hematopoietic growth factors (HGFs) introduced into induction chemotherapy (CT) of acute myeloid leukemia (AML) might be of benefit to treatment outcome by at least two mechanisms. HGFs given on days simultaneously with CT might sensitize the leukemic cells and enhance their susceptibility to CT. HGFs applied after CT might hasten hematopoietic recovery and reduce morbidity or mortality. MATERIALS AND METHODS: We set out to evaluate the use of granulocyte-macrophage colony-stimulating factor (GM-CSF; 5 microg/kg) in a prospective randomized study of factorial design (yes or no GM-CSF during CT, and yes or no GM-CSF after CT) in patients aged 15 to 60 years (mean, 42) with newly diagnosed AML. GM-CSF was applied as follows: during CT only (+/-, n = 64 assessable patients), GM-CSF during and following CT (+/+, n = 66), no GM-CSF (-/ , n = 63), or GM-CSF after CT only (-/+, n = 60). RESULTS: The complete response (CR) rate was 77%. At a median follow-up time of 42 months, probabilities of overall survival (OS) and disease-free survival (DFS) at 3 years were 38% and 37% in all patients. CR rates, OS, and DFS did not differ between the treatment groups (intention-to-treat analysis). Neutrophil recovery (1.0 x 10(9)/L) and monocyte recovery were significantly faster in patients who received GM-CSF after CT (26 days v 30 days; neutrophils, P < .001; monocytes, P < .005). Platelet regeneration, transfusion requirements, use of antibiotics, frequency of infections, and duration of hospitalization did not vary as a function of any of the therapeutic GM-CSF modalities. More frequent side effects (eg, fever and fluid retention) were noted in GM-CSF-treated patients predominantly related to the use of GM-CSF during CT. CONCLUSION: Priming of AML cells to the cytotoxic effects of CT by the use of GM-CSF during CT or accelerating myeloid recovery by the use of GM-CSF after CT does not significantly improve treatment outcome of young and middle-aged adults with newly diagnosed AML. PMID- 9396404 TI - Response rate accuracy in oncology trials: reasons for interobserver variability. Groupe Francais d'Immunotherapie of the Federation Nationale des Centres de Lutte Contre le Cancer. AB - PURPOSE: We evaluated the impact of an evaluation committee (EC) on patients' overall response status in a large multicenter trial in oncology. We identified reasons for disagreements between investigators and the EC. MATERIALS AND METHODS: The Cancer Renal Cytokine (CRECY) study was a French multicenter trial that tested cytokine therapy in 489 patients with metastatic renal cell carcinoma. Objective response (OR) evaluation included medical imaging and was studied according to international guidelines. A blinded peer review of all responders and litigious cases was performed by an EC. RESULTS: Major disagreements occurred in 40% and minor disagreements in 10.5% of the reviewed files. The number of significant tumor responses was reduced by 23.2% after review by the EC. Reasons for disagreements included errors in tumor measurements, errors in selection of measurable targets, intercurrent diseases, and radiologic technical problems. These reasons for disagreements are analyzed and discussed. CONCLUSION: We conclude that all therapeutic trial results should be reviewed by peer analysis of all presumed responders by an EC. International guidelines for response evaluation should be updated by including more reliable methods of measurements and definition of minimal imaging procedures. PMID- 9396405 TI - Breast cancer associated with palmar fasciitis and arthritis. PMID- 9396406 TI - Socioeconomic status versus survival in Ontario. PMID- 9396407 TI - Simplified regimen for the prevention of paclitaxel-associated hypersensitivity reactions. PMID- 9396408 TI - Does coronary endothelial dysfunction cause myocardial ischemia in the absence of obstructive coronary artery disease? PMID- 9396409 TI - When 'normal' cholesterol levels injure the endothelium. PMID- 9396410 TI - Selectins: vital vasculotropic vectors involved in vascular remodeling. PMID- 9396411 TI - Body iron stores and atherosclerosis. PMID- 9396412 TI - Oxidative stress and cardiovascular disease. PMID- 9396413 TI - Transient triglyceridemia decreases vascular reactivity in young, healthy men without risk factors for coronary heart disease. AB - BACKGROUND: Hypertriglyceridemia is now accepted as a risk factor for coronary heart disease, although the mechanism behind the increased risk is not well understood. The present study was undertaken to investigate the effects of triglyceridemia on endothelial function, because impaired endothelial function is considered a marker of atherogenesis. METHODS AND RESULTS: Flow- and nitroglycerin-induced dilatation of the brachial artery was investigated noninvasively by high-resolution ultrasound technique in seven young, healthy men without risk factors for coronary heart disease. Transient triglyceridemia was induced by infusion of a triglyceride emulsion, Intralipid, which raised free fatty acid concentrations twofold and triglyceride levels fourfold. Flow-induced vasodilatation decreased from 7.1+/-3.0% to 1.6+/-2.6% (P<.0002), whereas nitroglycerin-induced vasodilatation decreased from 20.5+/-5.8% to 11.5+/-3.2% (P<.002) before and after 1 hour of infusion of Intralipid, respectively. CONCLUSIONS: Transient triglyceridemia decreases vascular reactivity, presumably by both endothelium-dependent and endothelium-independent mechanisms. PMID- 9396414 TI - Impact of respiratory maneuvers on cardiac volume within left-breast radiation portals. AB - BACKGROUND: Late cardiac morbidity and mortality have been reported among left breast cancer survivors treated with radiation therapy. Radiation-induced cardiotoxicity is affected by the volume of myocardium included in the radiation portals. We hypothesize that simple respiratory maneuvers may alter the position of the heart relative to the portals without altering the radiation dose delivered to the breast. METHODS AND RESULTS: Fourteen healthy female adult volunteers underwent cardiac MRI to determine the cardiac volume included in the typical left-breast radiation field during respiratory maneuvers. Cardiac volume within the radiation portals was assessed from a transverse stack of 14 1-cm thick contiguous slices covering the entire heart, obtained during breath holding at end-tidal volume (baseline), deep inspiration, and forced expiration. Thirteen subjects (92%) had inclusion of a portion of the heart within the radiation portals at end-tidal volume (median, 20.9 cm3; range, 1.3 to 88.4 cm3). In these subjects, inspiration decreased the cardiac volume included within the radiation portals (median change: -10.7 cm3 [-40.2%], P<.001 versus end-tidal volume), whereas expiration increased the cardiac volume included (median change: 4.0 cm3 [21.5%]; P<.001 versus end-tidal volume). CONCLUSIONS: Inclusion of a portion of the heart in the left-breast radiation field is common. The use of simple inspiratory maneuvers significantly decreases cardiac volume within the radiation portals. Such an approach during delivery of radiation therapy may allow for preservation of radiation dosage to the breast while reducing cardiac involvement and subsequent mortality. PMID- 9396415 TI - Evaluation of different ventricular pacing sites in patients with severe heart failure: results of an acute hemodynamic study. AB - BACKGROUND: Multisite ventricular pacing has recently been proposed as an additional treatment for patients with severe congestive heart failure. To further assess the potential value of this technique, we compared the acute hemodynamic changes associated with pacing the right ventricular apex (RVA) or outflow tract (RVOT) alone, the left ventricle (LV) alone, or biventricular (BIV) pacing of the RVA and LV together. METHODS AND RESULTS: Acute hemodynamic findings were measured in 27 patients with severe heart failure despite optimal therapy and either first-degree AV block and/or an intraventricular conduction defect. In the 23 patients with a high pulmonary capillary wedge pressure (PCWP) (>15 mm Hg), data were collected after transvenous pacing at different ventricular sites in either the VDD mode (AV delay=100 ms) or the VVI mode in patients with atrial fibrillation (n=6). The mean baseline cardiac index was 1.82 L x min(-1) x m(-2). Mean+/-SD baseline systolic blood pressure (SBP) (118.5+/ 15.2 mm Hg), PCWP (26.4+/-6.6 mm Hg), and V-wave amplitude (39.1+/-14.6 mm Hg) were similar before and after either RVA or RVOT pacing. In contrast, LV-based pacing (either LV alone or BIV pacing) resulted in higher SBP (P<.03) and lower PCWP (P<.01) and V-wave amplitude (P<.001) than either baseline or RV pacing measurements. With LV pacing alone, SBP, PCWP, and V waves were 126.5+/-15.1, 20.7+/-5.9, and 25.5+/-8.1 mm Hg, respectively. The results with LV pacing alone were similar to those obtained with BIV pacing. CONCLUSIONS: In patients with severe congestive heart failure, both LV pacing alone and BIV pacing resulted in a similar and significant acute improvement in SBP, PCWP, and V-wave amplitude compared with baseline measurements and RV pacing alone. These results provide a strong basis for initiating long-term studies examining the chronic effects of LV based pacing in patients with medically refractory congestive heart failure. PMID- 9396416 TI - Changing features of anginal pain after PTCA suggest a stenosis on a different artery rather than restenosis. AB - BACKGROUND: We recently found that patients who had had two myocardial infarctions in different myocardial regions frequently reported different locations of infarct pain, whereas patients who had had two infarcts at the same site had a similar distribution of pain. The aim of this study was to assess whether a different location of anginal pain may help identify patients with a new stenosis on an artery perfusing another myocardial region as opposed to those with restenosis after coronary angioplasty (PTCA). METHODS AND RESULTS: We studied 38 patients (59+/-11 years old) who underwent PTCA for single-vessel disease, with recurrence of symptoms requiring repeat coronary angiography during a 3-year follow-up. According to our inclusion criteria, angiography showed either a significant restenosis of the dilated lesion, with no evidence of lesions in the other vessels (n=26), or a new stenosis in either of the other coronary arteries, with no restenosis in the dilated vessel (n= 12). Before each procedure, patients reported the location and radiation of anginal pain. There was no relation between location of pain and site of the coronary stenosis. However, none of the patients with restenosis reported a different location of pain after angioplasty, compared with 5 patients with new stenosis (0% versus 42%, P=.002). Radiation of pain involved different areas of the body in 1 patient with restenosis and in 6 with new stenosis (4% versus 50%, P=.002). Overall, location or radiation of pain in a different body area had a specificity of 96% and a sensitivity of 58% in detecting a stenosis on a new artery. CONCLUSIONS: A different location of anginal pain may distinguish patients with a new coronary stenosis from those with restenosis after PTCA for single-vessel disease. These findings suggest that in individual patients, differences in the location of cardiac pain may be indicative of the occurrence of ischemia in different myocardial regions. PMID- 9396417 TI - Coronary artery disease and polymorphisms in a receptor mediating shear stress dependent platelet activation. AB - BACKGROUND: Platelets play pivotal roles in coronary thrombosis, and antiplatelet therapies are widely used for coronary artery disease (CAD). However, the effects of genetic variation in platelets on CAD are poorly understood. We have assessed the association between CAD and polymorphisms in a platelet receptor for von Willebrand factor, the glycoprotein (GP) Ib/IX complex, which mediates shear stress-dependent platelet activation. METHODS AND RESULTS: Genotypes of the alpha chain of the receptor (GP Ib alpha, 145Thr/Met) were determined in 91 patients with myocardial infarction (MI) or angina pectoris whose lesions were confirmed by coronary angiography as well as in 105 individuals from the general population with no history of angina or other heart diseases and normal resting ECGs. There was no homozygote for Met/Met in either the control or patient groups. The prevalence of the Thr/Met genotype (T/M) in all patients was not significantly different from that in the control group. However, the frequency of T/M was significantly higher in patients aged < or = 60 years (31.8%) than in control subjects aged < or = 60 years (16.0%; P<.05, odds ratio=2.5). An association was also demonstrated between CAD and the other polymorphism of GP Ib alpha, a variable number of tandem repeats of a 13-amino acid sequence, which is known to be linked to the 145Thr/Met polymorphism. There was an association between the frequency of the T/M genotype and the angiographic severity of CAD: 11.1% for Gensini score < 40 versus 50.0% for Gensini score > or = 40 (P=.0015). There was no difference in the distribution of GP Ib alpha genotypes between patients with MI and those with angina pectoris. CONCLUSIONS: This study suggests that the presence of the Met allele in GP Ib alpha is a risk factor for the prevalence and severity of CAD in individuals aged < or = 60 years. The results need to be confirmed in a large-scale study of incident case subjects and matching control subjects. PMID- 9396418 TI - Endothelial dysfunction is associated with cholesterol levels in the high normal range in humans. AB - BACKGROUND: The purpose of this study was to test the hypothesis that cholesterol levels in the high normal range are associated with impaired endothelium dependent vasodilation. METHODS AND RESULTS: We studied leg blood flow (LBF) responses to graded intrafemoral artery infusions of the endothelium-dependent vasodilator methacholine chloride (MCh) or the endothelium-independent vasodilator sodium nitroprusside (SNP) in normal volunteers exhibiting a wide range of total cholesterol levels within the normal range (<75th percentile). LBF increased in a dose-dependent fashion in response to the femoral artery infusions of MCh and SNP (P<.001). LBF responses to MCh were significantly blunted (P<.001) in subjects with high normal cholesterol (195+/-6 mg/dL, n=13) compared with subjects with low normal cholesterol (146+/-5 mg/dL, n=20). Maximal endothelium dependent vasodilation in the high normal group was decreased by nearly 50% compared with the low normal group (146+/-13% versus 268+/-34%, P<.01). There was a negative correlation between total cholesterol levels and maximal endothelium dependent vasodilation (total cholesterol, r=-.41, P<.02; LDL cholesterol, r= .42, P<.02). On the other hand, LBF responses to the endothelium-independent vasodilator SNP did not differ between groups. CONCLUSIONS: These data suggest that an inverse and continuous relationship exists between the prevailing cholesterol level and endothelium-dependent vasodilation. Moreover, cholesterol levels even in the normal range may be associated with endothelial dysfunction, thus potentially contributing to the increased risk of macrovascular disease conferred by cholesterol elevations. PMID- 9396419 TI - Cardiovascular death and left ventricular remodeling two years after myocardial infarction: baseline predictors and impact of long-term use of captopril: information from the Survival and Ventricular Enlargement (SAVE) trial. AB - BACKGROUND: We quantified cardiovascular death and/or left ventricular (LV) dilatation in patients from the SAVE trial to determine whether dilatation continued beyond 1 year, whether ACE inhibitor therapy attenuated late LV dilatation, and whether any baseline descriptors predicted late dilatation. METHODS AND RESULTS: Two-dimensional echocardiograms were obtained in 512 patients at 11+/-3 days and 1 and 2 years postinfarction to assess LV size, percentage of the LV that was akinetic/dyskinetic (%AD), and LV shape index. LV function was assessed by radionuclide ejection fraction. Two hundred sixty-three patients (51.4%) sustained cardiovascular death and/or LV diastolic dilatation; 279 (54.5%) had cardiovascular death and/or systolic dilatation. In 373 patients with serial echocardiograms, LV end-diastolic and end-systolic sizes increased progressively from baseline to 2 years (both P<.01). More patients with LV dilatation had a decrease in ejection fraction: 24.8% versus 6.8% (P<.001) (diastole) and 25.7% versus 5.3% (P<.001) (systole). Captopril attenuated diastolic LV dilatation at 2 years (P=.048), but this effect was carried over from the first year of therapy because changes in LV size with captopril beyond 1 year were similar to those with placebo. Predictors of cardiovascular death and/or dilatation were age (P=.023), prior infarction (P<.001), lower ejection fraction (P<.001), angina (P=.007), heart failure (P=.002), LV size (P<.001), and infarct size (%AD) (P<.001). CONCLUSIONS: Cardiovascular death and/or LV dilatation occurred in >50% of patients by 2 years. LV dilatation is progressive, associated with chamber distortion and deteriorating function that is unaffected by captopril beyond 1 year. PMID- 9396420 TI - Body iron stores and the risk of carotid atherosclerosis: prospective results from the Bruneck study. AB - BACKGROUND: Fe2+ released from tissue iron stores may accelerate lipid peroxidation by virtue of its pro-oxidant properties and thus promote early atherogenesis. METHODS AND RESULTS: The present prospective survey addresses the potential association between serum ferritin concentrations and the 5-year progression of carotid atherosclerosis as assessed by ultrasonographic follow-up evaluations. The study population comprises a random sample of 826 men and women 40 to 79 years old. Serum ferritin was one of the strongest risk predictors of overall progression of atherosclerosis. The main part of this association appeared to act through modification of the atherogenic potential of LDL cholesterol (OR [95% CI] for a 1-SD unit increase in ferritin at LDL levels of 2.5, 3.6, and 4.9 mmol/L: 1.55 [1.30 to 1.85], 1.77 [1.40 to 2.24], and 2.05 [1.50 to 2.80]; P=.0012 for effect modification). Changes in iron stores during the follow-up period modified atherosclerosis risk, in that a lowering was beneficial and further iron accumulation exerted unfavorable effects. All these findings applied equally to incident atherosclerosis and the extension of preexisting atherosclerotic lesions. The significance of prominent iron stores in the development of carotid stenosis was clearly less pronounced. Finally, ferritin and LDL cholesterol showed a synergistic association with incident cardiovascular disease and death (n=59). CONCLUSIONS: The present study provided strong epidemiological evidence for a role of iron stores in early atherogenesis and suggests promotion of lipid peroxidation as the main underlying pathomechanism. This hypothesis could in part explain the sex difference in atherosclerotic vascular disease. PMID- 9396421 TI - Estimating effectiveness of cardiac arrest interventions: a logistic regression survival model. AB - BACKGROUND: The study objective was to develop a simple, generalizable predictive model for survival after out-of-hospital cardiac arrest due to ventricular fibrillation. METHODS AND RESULTS: Logistic regression analysis of two retrospective series (n=205 and n=1667, respectively) of out-of-hospital cardiac arrests was performed on data sets from a Southwestern city (population, 415,000; area, 406 km2) and a Northwestern county (population, 1,038,000; area, 1399 km2). Both are served by similar two-tiered emergency response systems. All arrests were witnessed and occurred before the arrival of emergency responders, and the initial cardiac rhythm observed was ventricular fibrillation. The main outcome measure was survival to hospital discharge. Patient age, initiation of CPR by bystanders, interval from collapse to CPR, interval from collapse to defibrillation, bystander CPR/collapse-to-CPR interval interaction, and collapse to-CPR/collapse-to-defibrillation interval interaction were significantly associated with survival. There was not a significant difference between observed survival rates at the two sites after control for significant predictors. A simplified predictive model retaining only collapse to CPR and collapse to defibrillation intervals performed comparably to the more complicated explanatory model. CONCLUSIONS: The effectiveness of prehospital interventions for out-of hospital cardiac arrest may be estimated from their influence on collapse to CPR and collapse to defibrillation intervals. A model derived from combined data from two geographically distinct populations did not identify site as a predictor of survival if clinically relevant predictor variables were controlled for. This model can be generalized to other US populations and used to project the local effectiveness of interventions to improve cardiac arrest survival. PMID- 9396422 TI - Increased formation of the isoprostanes IPF2alpha-I and 8-epi-prostaglandin F2alpha in acute coronary angioplasty: evidence for oxidant stress during coronary reperfusion in humans. AB - BACKGROUND: The role of oxidant stress in cardiac ischemia/reperfusion injury in humans remains controversial. This is due, in part, to the limitations of available indices of oxidant stress in vivo. Isoprostanes are stable, free radical-catalyzed products of arachidonic acid. We assessed their formation in patients undergoing coronary reperfusion via percutaneous transluminal coronary angioplasty (PTCA). METHODS AND RESULTS: We developed specific, mass spectrometry assays for two structurally distinct F2 isoprostanes, 8-epi-PGF2alpha and IPF2alpha-I. Urine samples for isoprostane determination were collected in patients undergoing coronary arteriography (n=11), elective PTCA (n=15), and angiography after thrombolysis for acute myocardial infarction (MI) (n=10). Urinary levels (pmol/mmol creatinine) of both isoprostanes were markedly increased from baseline in the first 6 hours after PTCA for acute MI (105+/-17.8 versus 230+/-66 for 8-epi-PGF2alpha [P=.009] and 466+/-91 versus 833+/-153 for IPF2alpha-I [P=.001]) and returned toward preprocedural values by 24 hours (122+/ 18 for 8-epi-PGF2alpha and 457+/-102 for IPF2alpha-I). There was a slight increase in urinary 8-epi-PGF2alpha levels (64.7+/-9.5 versus 84.9+/-10.6; P=.02) after diagnostic coronary arteriography and elective PTCA (88.7+/-7.5 versus 114.3+/-16.1; P=.01). A striking correlation was observed (r=.68, P<.0001; n=33) between urinary 8-epi-PGF2alpha and IPF2alpha-I levels in patients receiving thrombolytic agents for acute MI. CONCLUSIONS: Urinary F2 isoprostane levels are elevated in patients after treatments resulting in reperfusion for acute MI. These findings provide evidence consistent with increased oxidant stress in vivo in this setting. Measurement of urinary isoprostanes may offer a noninvasive approach to the assessment of oxidant stress and the efficacy of antioxidant therapies in these syndromes. PMID- 9396423 TI - Nonuniform nighttime distribution of acute cardiac events: a possible effect of sleep states. AB - BACKGROUND: Although 250,000 myocardial infarctions and 38,000 sudden cardiac deaths occur at night annually, this public health problem is underappreciated and poorly understood. We examined whether the incidence of myocardial infarction, sudden cardiac death, and automatic implantable cardioverter defibrillator (AICD) discharge was nonuniform, a result that may implicate physiological triggers such as sleep-state dependent changes in autonomic nervous system activity. METHODS AND RESULTS: We conducted a review of the circadian pattern of the onset of myocardial infarction (n=19), sudden cardiac death (n=12), and AICD discharge (n=7). The nighttime period was chosen a priori as midnight to 5:59 AM. These reports documented 11,633 nocturnal myocardial infarctions (20% of the total myocardial infarctions), 1981 nocturnal sudden cardiac deaths (14.6% of the total sudden cardiac deaths), and 1200 nocturnal AICD discharges (15.0% of the total discharges). The distributions of myocardial infarction, sudden cardiac death, and AICD discharge were each significantly nonuniform (P<.001). The peak incidence of myocardial infarction and AICD discharge occurred between midnight and 0:59 AM, whereas the peak incidence of sudden cardiac death was between 1:00 and 1:59 AM. The trough in incidence occurred between 4:00 and 4:59 AM for sudden cardiac death and between 3:00 and 3:59 AM for myocardial infarction and AICD discharge. CONCLUSIONS: Nocturnal myocardial infarction, sudden cardiac death, and AICD discharge exhibit nonuniform distributions. This finding is consistent with the hypothesis that sleep-state dependent fluctuations in autonomic nervous system activity may trigger the onset of major cardiovascular events and provides further impetus for more directly testing this hypothesis at population, individual, and mechanistic levels. A better understanding of nocturnal triggers may make it possible to reduce the incidence of myocardial infarction, ventricular tachyarrhythmias, and sudden cardiac death during the nocturnal period. PMID- 9396424 TI - Upregulation of angiotensin-converting enzyme during the healing process after injury at the site of percutaneous transluminal coronary angioplasty in humans. AB - BACKGROUND: Balloon injury models in rat have shown enhanced expression of ACE in the developing neointima. However, neointimal lesions in human coronary arteries are complex due to atherosclerosis and different types of wall laceration. This study was designed to investigate whether ACE is present in the neointima of humans, including patients with restenosis after percutaneous transluminal coronary angioplasty (PTCA). METHODS AND RESULTS: Thirty-seven sites with angioplasty injury, obtained at autopsy, were studied using immunocytochemical techniques. Sites with injury limited to a fibrous plaque and those with injury extending into the media (<2 months after PTCA) showed fibrocellular repair tissue composed mainly of smooth muscle cells that were distinctly positive for ACE. In cellular reactions at the site of injury limited to the atheromatous plaque (<2 months after PTCA), the expression of ACE appeared first in accumulated macrophages; once smooth muscle cells appeared in the repair tissue, they also expressed ACE. At a later stage (3 months after PTCA), the number of cells with ACE expression decreased markedly; from 7 months on, ACE was no longer expressed within the repair tissue. Basically, there were no differences with regard to ACE expression during the healing process after PTCA between segments with and those without angiographic evidence of restenosis. CONCLUSIONS: These results show that PTCA injury in humans results in upregulation of ACE at sites of active repair and, therefore, ACE could play an important role as one of the mediators of the healing process after PTCA. PMID- 9396425 TI - Albumin excretion rate increases during acute myocardial infarction and strongly predicts early mortality. AB - BACKGROUND: This study was undertaken to assess whether albumin excretion rate (AER) increases during acute myocardial infarction (AMI) and whether it predicts in-hospital mortality. METHODS AND RESULTS: The study was carried out in 496 subjects admitted to hospital for suspected AMI. Of these, 360 had evidence of AMI. The other 136 were studied as control subjects. AER was assessed by radioimmunoassay in three 24-hour urine collections performed on the first, third, and seventh days after admission. Left ventricular ejection fraction was measured by two-dimensional echocardiography in 254 subjects. AER adjusted for several confounders was higher in the AMI than the non-AMI group on the first (69.2+/-5.2 versus 27.3+/-8.5 mg/24 h, P<.0001) and third (30.3+/-2.7 versus 12.5+/-4.4 mg/24 h, P=.001) days, whereas no difference was present on the seventh day. When the subjects with heart failure were excluded, the difference between the two groups remained significant (first day, P<.0001; third day, P=.001). On the basis of classification of the 26 AMI patients who died in hospital according to whether they had normal AER, microalbuminuria, or overt albuminuria, mortality rate progressively increased with increasing levels of AER (P<.0001). In a Cox's proportional hazards model, AER was a better predictor of in-hospital mortality than Killip class or echocardiographic left ventricular ejection fraction. A cutoff value of 50 mg/24 h for first-day AER and 30 mg/24 h for third-day AER yielded a sensitivity of 92.3% and of 88.5% and a specificity of 72.4% and of 79.3%, respectively, for mortality. Adjusted relative risks for the two cutoff values were 17.3 (confidence limits, 4.6 to 112.7) and 8.4 (confidence limits, 2.4 to 39.3), respectively. CONCLUSIONS: These data show that AER increases during AMI and that it yields prognostic information additional to that provided by clinical or echocardiographic evaluation of left ventricular performance. PMID- 9396426 TI - Effects of dobutamine at maximally tolerated dose on myocardial blood flow in humans with ischemic heart disease. AB - BACKGROUND: This study tests the hypothesis in humans with ischemic heart disease that myocardial blood flow response to dobutamine is linearly correlated with blood flow response to adenosine. METHODS AND RESULTS: PET with [13N]ammonia was used to measure myocardial blood flow at rest and during adenosine and dobutamine at the maximally tolerated dose. Myocardial segments were defined physiologically on the basis of blood flow response to adenosine: normal, > or = 2 mL x min(-1) x g(-1); abnormal, < 2 mL x min(-1) x g(-1); and "steal," decline versus baseline > or = 0.15 mL x min(-1) x g(-1). The patient population consisted of 11 men and 2 women. Dobutamine increased heart rate (79+/-22 to 115+/-28 bpm) and rate pressure product (9748+/-2862 to 15,157+/-3433 mm Hg/min) significantly (both P<.01). Myocardial blood flow at rest in abnormal segments (0.50+/-0.23 mL x min( 1) x g(-1)) was reduced (P<.001) versus normal (0.90+/-0.45) and steal (0.92+/ 0.60). Nevertheless, in abnormal segments, blood flow increased versus rest (P<.001) with dobutamine (0.83+/-0.43) and adenosine (0.90+/-0.49). In steal segments, myocardial blood flow declined versus baseline (P<.001) with dobutamine (0.68+/-0.46) and adenosine (0.50+/-0.45). In normal segments, myocardial blood flow increased (P<.001) with dobutamine (2.16+/-0.99) and adenosine (3.10+/ 0.90). Over the range of flows, the correlation between adenosine and dobutamine was good (r=.78, P<.0001). Although flow with dobutamine in normal segments correlated with rate-pressure product (r=.81, P<.05), the slope of the line was 2.7+/-0.8 (P<.02), and normalized blood flow (3.3+/-2.5 x rest) exceeded normalized rate-pressure product (1.9+/-0.8 x rest; P<.05). CONCLUSIONS: In humans with ischemic heart disease, myocardial blood flow responses to dobutamine and adenosine are linearly correlated over a wide range. The hyperemic response to dobutamine is in excess of that predicted by rate-pressure product and reflects the unmeasured inotropic, oxygen-wasting, and beta2-agonist effects of the drug. Dobutamine induces coronary steal with a frequency approaching that of adenosine. PMID- 9396427 TI - Influence of infarct-zone viability on left ventricular remodeling after acute myocardial infarction. AB - BACKGROUND: The relation between residual myocardial viability after acute myocardial infarction (AMI) and ventricular remodeling has yet to be fully elucidated. We hypothesized that the presence of residual viability would favorably influence left ventricular remodeling after AMI and that serial changes in left ventricular dimensions might be related to the extent of myocardial viability in the infarct zone. METHODS AND RESULTS: Ninety-three patients with a first AMI successfully treated with primary coronary angioplasty underwent two dimensional echocardiography within 24 hours of admission and low-dose dobutamine echocardiography at a mean of 3 days after AMI. Two-dimensional echocardiography and coronary angiography were obtained in all patients 1 and 6 months after coronary angioplasty. On the basis of dobutamine echocardiography responses, patients were divided in two subsets: those with (n=48; group I) and those without (n=45; group II) infarct-zone viability. There was no difference in minimal lesion diameter and infarct-related artery patency at 1 and 6 months between the two groups. Group II patients had significantly greater end-diastolic (76+/-18 versus 53+/-14 mL/m2; P<.0003) and end-systolic (42+/-17 versus 22+/-11 mL/m2; P<.0003) volumes at 6 months than did patients in group 1. The extent of infarct-zone viability was significantly inversely correlated with percent changes in end-diastolic volumes at 6 months (r=-.66; P<.000001) and was the most powerful independent predictor of late left ventricular dilation. CONCLUSIONS: After reperfused AMI, the degree of left ventricular dilation, when it occurs, is inversely related to the extent of residual myocardial viability in the infarct zone. Thus, the absence of residual infarct-zone viability discriminates patients who develop progressive left ventricular dilation after reperfused AMI from those who maintain normal left ventricular geometry. PMID- 9396428 TI - Topographic analysis of proliferative activity in carotid endarterectomy specimens by immunocytochemical detection of the cell cycle-related antigen Ki 67. AB - BACKGROUND: On the basis of contradictory results found in animal experiments and coronary atherectomy tissue, there is an ongoing debate about the significance of cellular proliferation in human atherosclerosis. In the present prospective study, the cell cycle-related antigen Ki-67 was detected for topographic determination of cell turnover in distinct regions of human carotid endarterectomy specimens harvested en bloc by surgical biopsy. METHODS AND RESULTS: After en bloc resection, serial sections of 26 consecutive carotid lesions were analyzed by histomorphological examination and immunohistochemistry. Thereby, 319 high-power fields were attributed to separate plaque regions defined as follows: distal boundary of the lesion with normal intima, plaque shoulder, core region, and diffuse intimal thickening. Endothelial cells, smooth muscle cells, T cells, and macrophages were identified by immunostaining of factor VIII related protein, alpha-actin, CD68, and CD45R0. An overall proliferation index of 0.49+/-1.05% was yielded by positive anti-Ki-67 immunolabeling, predominantly in macrophage-rich areas characterized by high cell density (>1000 cells/mm2) as well as in reparative sites in the perimeter of atheroma, intramural thrombosis, plaque hemorrhage, and neovascularization (P<.01). Few or no signs of proliferation activity were found in normal intima, in areas of dense alpha-actin positivity, or adjacent media. As shown by double immunostaining, macrophages and unspecified mesenchymal cells represented the prevailing proliferating cell type. CONCLUSIONS: Our results suggest that proliferation in advanced human carotid lesions is confined to the intima and focally concentrated in central plaque regions negative for alpha-actin. Furthermore, it apparently occurs primarily as part of inflammatory processes and structural repair predominantly involving macrophages, as well as unspecific mesenchymal cells. PMID- 9396430 TI - Endothelium-dependent dilatation is impaired in young healthy subjects with a family history of premature coronary disease. AB - BACKGROUND: A family history of premature coronary artery disease (CAD) in a first-degree relative is an independent risk factor for coronary disease. Both genetic and environmental influences are likely to be responsible and may interact, but their relative importance is unclear. METHODS AND RESULTS: We studied endothelial function in 50 first-degree relatives (31 men, 19 women; mean age, 25+/-8 years) of patients (men < or = 45 years, women < or = 55 years) with proven CAD. All subjects were well, lifelong nonsmokers, not diabetic, and not hypertensive and took no medications. Using high-resolution external vascular ultrasound, we measured brachial artery diameter at rest and in response to reactive hyperemia (with increased flow causing an endothelium-dependent vasodilatation) and to sublingual glyceryltrinitrate (GTN, an endothelium independent dilator). Vascular responses were compared with those of 50 healthy control subjects matched for age and sex. Flow-mediated dilatation (FMD) was impaired in the family history group (4.9+/-4.6% versus 8.3+/-3.5% in control subjects, P<.005). In contrast, GTN caused dilatation in all subjects (family history, 17.1+/-8.8%; control subjects, 19.0+/-6.3%; P=NS), suggesting that reduced FMD was due to endothelial dysfunction. When the family history subjects were subdivided, those found to have a serum cholesterol > 4.2 mmol/L (group A, n=10) had mildly impaired FMD compared with control subjects (5.5+/-5.1% versus 8.3+/-3.5%). In others whose affected relative had coronary risk factors (group B, n=24), FMD was also only slightly reduced (6.2+/-4.8% versus 8.3+/-3.5%). In contrast, subjects with no risk factors and whose affected relative had a normal cardiovascular risk factor profile (group C, n=16) had markedly impaired FMD (2.9+/-3.7% versus 8.3+/-3.5%). Although ANOVA of the three family history subgroups did not reach statistical significance (F=2.55, P=.09), pairwise analysis showed that FMD in group C was significantly impaired compared with group B (P=.026). CONCLUSIONS: Healthy young adults with a family history of premature coronary disease may have impaired endothelium-dependent dilatation, even in the absence of other cardiovascular risk factors. Those subjects, who were free of risk factors and whose affected first-degree relative was free of risk factors, had the most impaired endothelial function, suggesting a genetic influence on early arterial physiology that may be relevant to later clinical disease. PMID- 9396429 TI - Prognostic value of intracoronary flow velocity and diameter stenosis in assessing the short- and long-term outcomes of coronary balloon angioplasty: the DEBATE Study (Doppler Endpoints Balloon Angioplasty Trial Europe). AB - BACKGROUND: The aim of this prospective, multicenter study was the identification of Doppler flow velocity measurements predictive of clinical outcome of patients undergoing single-vessel balloon angioplasty with no previous Q-wave myocardial infarction. METHODS AND RESULTS: In 297 patients, a Doppler guidewire was used to measure basal and maximal hyperemic flow velocities proximal and distal to the stenosis before and after angioplasty. In 225 patients with an angiographically successful percutaneous transluminal coronary angioplasty (PTCA), postprocedural distal coronary flow reserve (CFR) and percent diameter stenosis (DS%) were correlated with symptoms and/or ischemia at 1 and 6 months, with the need for target lesion revascularization, and with angiographic restenosis (defined as DS > or = 50% at follow-up). Logistic regression and receiver operator characteristic curve analyses were applied to determine the prognostic cutoff value of CFR and DS separately and in combination. Optimal cutoff criteria for predictors of these clinical events were DS, 35%; CFR, 2.5. A distal CFR after angioplasty > 2.5 with a residual DS < or = 35% identified lesions with a low incidence of recurrence of symptoms at 1 month (10% versus 19%, P=.149) and at 6 months (23% versus 47%, P=.005), a low need for reintervention (16% versus 34%, P=.024), and a low restenosis rate (16% versus 41%, P=.002) compared with patients who did not meet these criteria. CONCLUSIONS: Measurements of distal CFR after PTCA, in combination with DS%, have a predictive value, albeit modest for the short- and long-term outcomes after PTCA, and thus may be used to identify patients who will or will not benefit from additional therapy such as stent implantation. PMID- 9396431 TI - Silent myocardial ischemia in Kawasaki disease: evaluation of percutaneous transluminal coronary angioplasty by dobutamine stress testing. AB - BACKGROUND: Myocardial ischemia and myocardial infarction are the most serious complications of coronary artery lesions in children with Kawasaki disease (KD). Therefore, early detection and treatment of myocardial ischemia in patients with KD is essential. We studied the effectiveness of percutaneous transluminal coronary angioplasty (PTCA) in patients with silent myocardial ischemia detected by dobutamine stress 99mTc myocardial scintigraphy (TMS), body surface mapping (BMS), and signal-averaged ECG late potentials (ELP). METHODS AND RESULTS: Eight of 76 asymptomatic patients with a coronary stenosis >25% and a positive dobutamine stress test were considered to have silent myocardial ischemia. All eight patients had >95% stenoses demonstrated by coronary angiography (CAG) just before PTCA. After PTCA, CAG showed that all of the coronary artery stenoses had been reduced to <50%. Additionally, intravascular ultrasonography (IVUS) performed in five patients before and after PTCA demonstrated adequate dilation of the coronary stenosis after PTCA. All eight patients underwent dobutamine stress TMS, BMS, and ELP 2 to 3 months after PTCA, which demonstrated no regions of myocardial ischemia. Approximately 6 months later, CAG was performed in all eight patients, and only one patient had developed restenosis. CONCLUSIONS: PTCA effectively dilates stenotic coronary arteries in children with KD. Moreover, dobutamine stress TMS, BMS, and ELP are useful for detecting silent myocardial ischemia and estimating the effectiveness of PTCA. Furthermore, IVUS is useful for evaluating the severity of coronary artery lesions before and after PTCA in patients with KD. PMID- 9396432 TI - Coronary endothelial dysfunction in humans is associated with myocardial perfusion defects. AB - BACKGROUND: Coronary endothelial dysfunction may occur in patients with minimally obstructive coronary artery disease and angina, and potentially may cause myocardial ischemia. METHODS AND RESULTS: Coronary endothelium-dependent vasodilation was examined in patients with angina and <50% coronary artery diameter (CAD) stenosis by selectively infusing acetylcholine (10(-6) mol/L to 10(-4) mol/L) into the left anterior descending coronary artery (LAD). Percent change in CAD (%deltaCAD) was measured by quantitative coronary angiography, and percent change in coronary blood flow (%deltaCBF) was calculated using intracoronary flow Doppler. Coronary endothelium-independent vasodilation was examined using intracoronary adenosine and nitroglycerin. 99mTc sestamibi was injected intravenously just prior to the infusion of the highest dose of acetylcholine. Patients were divided blindly into three groups: Perfusion defects in non-LAD territory (group 1, n=6), no perfusion defects (group 2, n=7), and perfusion defects in the LAD territory (group 3, n=7). All patients had intact endothelium-independent vasodilation. In group 1, perfusion defects outside the LAD territory reflected an increase in %deltaCAD and %deltaCBF by 24+/-5% and 241+/-46% in the LAD. In group 2, %deltaCAD decreased by 26+/-5%, but %deltaCBF increased by 54+/-17%. In group 3, perfusion defects were within the LAD territory, reflecting a decrease in %deltaCAD and %deltaCBF by 35+/-5% and 51+/ 14%, respectively. CONCLUSIONS: This study demonstrates that coronary endothelial dysfunction in humans may be temporally associated with myocardial perfusion defects and supports a role for the coronary epicardial and microcirculation endothelium in regulating myocardial perfusion. Myocardial ischemia may occur in humans with impaired endothelium-dependent coronary flow reserve of the coronary epicardial and microcirculation. PMID- 9396433 TI - Effect of nadroparin, a low-molecular-weight heparin, on clinical and angiographic restenosis after coronary balloon angioplasty: the FACT study. Fraxiparine Angioplastie Coronaire Transluminale. AB - BACKGROUND: Experimental studies suggest that the antiproliferative effect of heparin after arterial injury is maximized by pretreatment. No previous studies of restenosis have used a pretreatment strategy. We designed this study to determine whether treatment with nadroparin, a low-molecular-weight heparin, started 3 days before the procedure and continued for 3 months, affected angiographic restenosis or clinical outcome after coronary angioplasty. METHODS AND RESULTS: In a prospective multicenter, double-blind, randomized trial, elective coronary angioplasty was performed on 354 patients who were treated with daily subcutaneous nadroparin (0.6 mL of 10,250 anti-Xa IU/mL) or placebo injections started 3 days before angioplasty and continued for 3 months. Angiography was performed just before and immediately after angioplasty and at follow-up. The primary study end point was angiographic restenosis, assessed by quantitative coronary angiography 3 months after balloon angioplasty. Clinical follow-up was continued up to 6 months. Clinical and procedural variables and the occurrence of periprocedural complications did not differ between groups. At angiographic follow-up, the mean minimal lumen diameter and the mean residual stenosis in the nadroparin group (1.37+/-0.66 mm, 51.9+/-21.0%) did not differ from the corresponding values in the control group (1.48+/-0.59 mm, 48.8+/ 18.9%). Combined major cardiac-related clinical events (death, myocardial infarction, target lesion revascularization) did not differ between groups (30.3% versus 29.6%). CONCLUSIONS: Pretreatment with the low-molecular-weight heparin nadroparin continued for 3 months after balloon angioplasty had no beneficial effect on angiographic restenosis or on adverse clinical outcomes. PMID- 9396434 TI - Aortic valve replacement in patients 80 years of age and older: survival and cause of death based on 1100 cases: collective results from the UK Heart Valve Registry. AB - BACKGROUND: Aging of the population and advances in preoperative and postoperative care are reflected in an increasing number of patients > or = 80 years of age undergoing aortic valve replacement (AVR) in the United Kingdom. The present study presents data on postoperative 30-day mortality, actuarial survival, and cause of death based on a large collective patient population. METHODS AND RESULTS: Data were extracted from the UK Heart Valve Registry. From January 1986 to December 1995, 1100 patients > or = 80 years of age underwent AVR and were reported to the registry. Six hundred eleven patients (55.5%) were women. The mean follow-up time was 38.9 months. The 30-day mortality was 6.6%. Of the 73 early deaths, 42 were due to cardiac reasons. The actuarial survival was 89%, 79.3%, 68.7%, and 45.8% at 1, 3, 5, and 8 years, respectively. After the first 30 postoperative days, 144 of the 205 deaths were due to noncardiac reasons. Malignancy, stroke, and pneumonia were the most common causes of late death. Bioprosthetic valves were implanted in 969 patients (88%) and mechanical valves in 131 (12%) patients. There was no difference in early mortality and actuarial survival between the two groups (P>.05). CONCLUSIONS: The above results suggest that under the selection criteria for AVR currently applied in the United Kingdom, patients > or = 80 years of age show a satisfactory early postoperative outcome and moderate medium-term survival benefit. PMID- 9396435 TI - Grading of mitral regurgitation by quantitative Doppler echocardiography: calibration by left ventricular angiography in routine clinical practice. AB - BACKGROUND: Quantitative Doppler echocardiography and proximal flow convergence methods are validated techniques for quantifying mitral regurgitation. However, the clinical interpretation of the values calculated is hindered by the absence of calibration of ranges of severity in large numbers of patients. METHODS AND RESULTS: In 180 consecutive patients (men, 62%; mean age+/-SD, 66+/-11 years), the results of Doppler quantification of isolated mitral regurgitation were calibrated by use of left ventricular angiographic grading performed within 3 months in routine practice and without intervening events. The thresholds of the quantitative variables corresponding to the angiographic grades were identified by maximizing the sum of sensitivity and specificity and minimizing their difference. The mitral regurgitation grade by angiography was 2.7+/-1.3. The mean value and correlation with angiographic grades for effective regurgitant orifice were 43+/-37 mm and r=.79 (P<.0001); for regurgitant volume, 62+/-45 mL and r=.80 (P<.0001); and for regurgitant fraction, 45+/-17% and r=.78 (P<.0001). Despite some overlap, differences between mitral regurgitation grades were all significant (all P<.05). The thresholds for severe mitral regurgitation (grade 4) were 60 mL, 50%, and 40 mm2 for regurgitant volume, regurgitant fraction, and orifice, respectively. CONCLUSIONS: In routine practice in large numbers of patients in a clinical laboratory, Doppler echocardiographic quantification of mitral regurgitation shows highly significant correlation with qualitative angiographic grades. Despite an expected overlap between classes, the calibration by angiography of grading ranges for the quantitative variables provides a framework for their interpretation and allows the definition in clinical practice of thresholds for severe mitral regurgitation. PMID- 9396436 TI - Increased myocardial muscarinic receptor density in idiopathic dilated cardiomyopathy: an in vivo PET study. AB - BACKGROUND: Congestive heart failure is associated with decreased stimulated myocardial adenylate cyclase activity, increased Gi-binding protein, attenuated parasympathetic tone, and increased modulation of beta-adrenergic inotropic left ventricular stimulation by parasympathetic agonists. Despite these abnormalities, changes in the density or affinity of ventricular muscarinic receptors have not been demonstrated in patients. METHODS AND RESULTS: The density and affinity constants of myocardial muscarinic receptors were evaluated noninvasively by means of positron emission tomography with 11C-MQNB (methylquinuclidinyl benzilate), a specific hydrophilic antagonist, in 20 patients with congestive heart failure due to idiopathic dilated cardiomyopathy (mean left ventricular ejection fraction, 22+/-9%) and compared with values in 12 normal subjects. The mean receptor concentration was significantly higher in patients than in control subjects (B'max, 34.5+/-8.9 versus 25+/-7.7 pmol/mL, P<.005), with no changes in affinity constants. The change in heart rate after injection of 0.6 mg of cold MQNB was lower in patients than in control subjects (34+/-20% versus 55+/-36%, P<.05), and receptor density correlated negatively with maximal heart rate in the patients (r=.45, P<.05). CONCLUSIONS: Congestive heart failure is associated with an upregulation of myocardial muscarinic receptors. This may be an adaptive mechanism to beta-agonist stimulation and should increase the number of potential targets for pharmacological intervention. PMID- 9396437 TI - Regional sympathetic nervous activity and oxygen consumption in obese normotensive human subjects. AB - BACKGROUND: Disturbed sympathetic nervous function may be of importance in obesity; sympathetic underactivity could contribute to deficient thermogenesis, positive energy balance, and weight gain, while in contrast, sympathetic nervous overactivity would predispose to the development of obesity-related hypertension. Global indices of sympathetic nervous system (SNS) function such as plasma or urinary norepinephrine (NE) have been unable to define SNS status in obesity. Since regional SNS activity can be altered in the absence of global changes, we investigated SNS activity in the heart, kidneys, and hepatomesenteric bed in healthy human subjects across a wide body mass index (BMI) range of between 19.6 and 35.5. METHODS AND RESULTS: Whole-body and regional plasma NE kinetics using [3H]-labeled NE were assessed. Regional oxygen consumption was measured by combining arteriovenous differences in oxygen content and regional blood flow. Arterial plasma NE and whole-body plasma NE spillover were unrelated to BMI. With a BMI cutoff of 27, mean cardiac NE spillover was 46% lower in the obese subjects when compared with the lean subjects (P=.017). Renal NE spillover was significantly correlated with BMI (r=.668, P=.001), the mean value in the obese subjects being more than twice that in the lean subjects. Hepatomesenteric NE spillover was comparable in lean and obese subjects. Renal and hepatomesenteric oxygen consumption were both significantly higher in the obese subjects compared with lean subjects. CONCLUSIONS: Regional SNS activity is heterogeneous in the obese state. Important regional alterations, which may be clinically relevant, occur in the absence of changes in global indices of sympathetic nervous function. The enhanced renal NE spillover in obesity may have implications for the development of hypertension in this group, whereas the low cardiac sympathetic tone would be expected to be cardioprotective. Enhanced visceral oxygen consumption evident in the kidneys and hepatomesenteric circulation in proportion to body mass contributes to the greater resting oxygen consumption in obesity. PMID- 9396438 TI - Left ventricular mechanics and geometry in patients with congenital complete atrioventricular block. AB - BACKGROUND: Radiographic evidence of cardiomegaly is common in patients with congenital complete atrioventricular block (CCAVB). It has been speculated that left ventricular (LV) remodeling and increased stroke volume counteract the bradycardia, but the effects of slow heart rate and atrioventricular asynchrony on LV dimensions, geometry, wall stress, and function have not been examined in detail. METHODS AND RESULTS: Thirty patients with CCAVB without associated congenital heart disease (mean age, 8.5+/-5.3 years; range, 0.2 to 20 years) were included in a cross-sectional two-institution study. Thirty-five echocardiograms were performed using standard techniques. ECG and 24-hour ECG recordings were reviewed. Seven patients did not receive a pacemaker, whereas 23 patients underwent pacemaker implantation after the echocardiogram. Compared with normal control subjects, LV volume (Z score=1.5+/-1.3) and LV mass (Z=1.2+/-1.5) were significantly increased, whereas LV mass-to-volume ratio (1.1+/-0.3) and geometry (short-axis diameter/length ratio=0.65+/-0.09) were normal. LV end-systolic stress (ESS) (a measure of afterload) was normal (Z score=0.2+/-2.3), whereas shortening fraction (Z=3+/-2.9) and velocity of circumferential fiber shortening (VCF) (Z=3+/-3.1) were increased. The relationship between VCF and ESS (a preload insensitive and afterload-adjusted index of contractility) was increased (Z=2.2+/ 2) with only small increase in preload (Z=1.02+/-1.1). Regression analyses showed no significant change over age in LV mass, volume, geometry, loading conditions, or systolic function. Patients who ultimately met criteria for pacemaker implantation did not differ from those who did not in terms of heart rate or LV function but did have increased LV volume (Z score=1.8+/-1.4 versus 0.4+/-0.9, P=.03) and LV mass (Z score=1.7+/-1.2 versus 0.2+/-1.7, P=.001) compared to the unpaced group. CONCLUSIONS: In most patients with CCAVB, the LV was enlarged with normal geometry and enhanced systolic function during the first two decades of life. The degree of LV dilation and enhanced function did not significantly change with age. In patients who ultimately underwent pacemaker implantation LV function did not differ from those who remained unpaced, but evidence of a slightly increased load manifested as increased end-diastolic volume and mass. PMID- 9396439 TI - Left ventricular contractile effects of inducible nitric oxide synthase in the human allograft. AB - BACKGROUND: Myocardial expression of inducible (i) nitric oxide (NO) synthase (iNOS) gene has been reported in transplant recipients and in dilated cardiomyopathy. NO derived from NO donor or from coronary endothelium has previously been shown in the human heart to reduce end-systolic left ventricular (LV) pressure, especially during beta-adrenoreceptor stimulation, because of earlier onset of LV relaxation. The present study investigated in transplant recipients whether a similar cardiodepressant effect could be attributed to NO derived from iNOS. METHODS AND RESULTS: In 16 transplant recipients who were free of rejection or graft vasculopathy, microtip LV pressure recordings, LV angiograms, and endomyocardial biopsies were obtained at annual coronary angiography. In 8 transplant recipients, microtip LV pressure recordings were obtained during intravenous dobutamine (5 microg x kg(-1) x min(-1)). Competitive reverse transcription-polymerase chain reaction of iNOS mRNA was performed on the endomyocardial biopsies, and the intensity of iNOS mRNA expression was quantified on a scale ranging from 0 to 5+. All measures of baseline LV function were comparable in transplant recipients with low (< or = 2+) or high myocardial iNOS mRNA. During intravenous dobutamine infusion, there was a significant correlation between the abbreviation of LV electromechanical systole time (LVEST is the time from onset of QRS to dP/dt(min)) and the rise of LV dP/dt(max) (r=.79; P<.02). By use of a multiple regression analysis, addition of the intensity of iNOS mRNA expression as an independent variable significantly (P<.005) improved the correlation between deltaLVEST and deltaLV dP/dt(max) (P<.001; r=.97), implying a larger abbreviation of LV contraction for a similar rise in LV dP/dt(max), when myocardial iNOS mRNA was higher. The larger abbreviation of LV contraction in patients with high iNOS mRNA was associated with a decrease in LV end-systolic pressure (-31+/-16 mm Hg). CONCLUSIONS: Myocardial iNOS gene expression in the human allograft influences the LV contractile response to beta-adrenergic stimulation through earlier onset of LV relaxation and reduction of LV end systolic pressure. These effects are similar to the LV contractile effects of NO derived from NO donor or from coronary endothelium. PMID- 9396440 TI - Percutaneous transluminal mitral valvuloplasty normalizes baroreflex sensitivity and sympathetic activity in patients with mitral stenosis. AB - BACKGROUND: In patients with mitral stenosis, reduced cardiac output or altered pulmonary hemodynamics may increase sympathetic nerve activity. However, the magnitude of the increase in sympathetic activity in such patients and the effect of valvuloplasty on this activity are unknown. METHODS AND RESULTS: We microneurographically measured muscle sympathetic nerve activity before and after mitral valvuloplasty in 10 patients (mean+/-SEM age, 48+/-2 years) with mitral stenosis and in 10 healthy volunteers (47+/-4 years); hemodynamic variables were also measured. Baroreflex sensitivity was assessed on the basis of the ratio of the change in heart rate or muscle sympathetic activity to the change in mean arterial pressure during intravenous infusion of sodium nitroprusside or phenylephrine. At baseline, muscle sympathetic activity was significantly higher in the patients with mitral stenosis than in the control subjects (42.1+/-3.2 versus 26.1+/-3.7 bursts/min, P<.05). However, there was no significant difference between the groups in sympathetic activity at 1 week after valvuloplasty. The reduction in sympathetic activity after valvuloplasty was maintained for > or = 6 months and correlated with the increase in cardiac index (r=.74, P<.05). Baroreflex sensitivity was significantly lower in the patients than in the control subjects, but after valvuloplasty there was no significant difference in baroreflex sensitivity between the groups. CONCLUSIONS: Sympathetic activity is increased in patients with mitral stenosis. Mitral valvuloplasty in such patients results in early and long-lasting normalization of sympathetic nerve activity, possibly because of an improvement in arterial baroreflex sensitivity. PMID- 9396441 TI - Arterial baroreflex modulation of heart rate in chronic heart failure: clinical and hemodynamic correlates and prognostic implications. AB - BACKGROUND: In chronic heart failure (CHF), arterial baroreflex regulation of cardiac function is impaired, leading to a reduction in the tonic restraining influence on the sympathetic nervous system. Because baroreflex sensitivity (BRS), as assessed by the phenylephrine technique, significantly contributes to postinfarction risk stratification, the aim of the present study was to evaluate whether in CHF patients a depressed BRS is associated with a worse clinical hemodynamic status and unfavorable outcome. METHODS AND RESULTS: BRS was assessed in 282 CHF patients in sinus rhythm receiving stable medical therapy (age, 52+/-9 years; New York Heart Association [NYHA] class, 2.4+/-0.6; left ventricular ejection fraction [LVEF], 23+/-6%). The BRS of the entire population averaged 3.9+/-4.0 ms/mm Hg (mean+/-SD) and was significantly related to LVEF and hemodynamic parameters (LVEF, P<.005; cardiac index and pulmonary wedge pressure, P<.001 by regression analysis). Patients in NYHA classes III or IV and those with severe mitral regurgitation had markedly depressed vagal reflexes. The association of BRS with survival was described after its categorization in three groups: below the lowest quartile (<1.3 ms/mm Hg), between the lowest quartile and the median (1.3 to 3 ms/mm Hg), and above the median (>3 ms/mm Hg). During a mean follow-up of 15+/-12 months, 78 primary events (cardiac death, nonfatal cardiac arrest, and status 1 priority transplantation) occurred (27.6%). BRS was significantly related to outcome (log rank, 9.1; P<.01), with a relative risk of 2.7 (95% confidence interval, 1.6 to 4.7) for patients with the major derangement in BRS (<1.3 ms/mm Hg). At multivariate analysis, BRS was an independent predictor of death after adjustment for noninvasive known risk factors but not when hemodynamic indexes were also considered. In CHF patients with severe mitral regurgitation, however, BRS remained a strong prognostic marker independent of hemodynamic function. CONCLUSIONS: In moderate to severe CHF, a depressed sensitivity of vagal reflexes parallels the deterioration of clinical and hemodynamic status and is significantly associated with poor survival. Particularly in patients with severe mitral regurgitation the baroreceptor modulation of heart rate provides prognostic information of incremental value to hemodynamic parameters. PMID- 9396442 TI - Knowledge of perfusion and contractile reserve improves the predictive value of recovery of regional myocardial function postrevascularization: a study using the combination of myocardial contrast echocardiography and dobutamine echocardiography. AB - BACKGROUND: This study was designed to determine the value of myocardial contrast echocardiography (MCE) and dobutamine echocardiography (DE), alone or in combination, in predicting functional recovery in patients with resting wall motion abnormalities due to CAD. MCE and DE have been independently shown to be useful in detecting myocardial viability in the post-myocardial infarction setting. METHODS AND RESULTS: Thirty-nine patients with significant coronary artery disease and resting wall motion abnormalities underwent DE (2.5 to 20 microg x kg(-1) x min(-1)) and wall motion analysis (16-segment model). MCE was performed with selective intracoronary injections of sonicated meglumine (2 cm3). Myocardial viability was defined as presence of contrast effect by MCE and contractile reserve or an ischemic response by DE. Functional recovery (improvement in wall motion) was assessed after revascularization (percutaneous transluminal coronary angioplasty, n=20; coronary artery bypass surgery, n=19). When the two groups of patients were analyzed, MCE was associated with excellent sensitivities (84%) yet poor specificities (19% to 26%); DE had lower sensitivities (79% to 80%) but also poor specificities (30% to 36%). The combination of both was associated with excellent sensitivities (90% to 93%) and modest specificities (48% to 50%) for predicting functional recovery. A biphasic response with DE was infrequent (14% to 42%) but highly specific of functional recovery (84% to 94%). MCE had an excellent negative predictive value for functional recovery (83%). CONCLUSIONS: The prediction of functional recovery post-revascularization can be enhanced by combining MCE and DE. PMID- 9396443 TI - Abundance and location of the small heat shock proteins HSP25 and alphaB crystallin in rat and human heart. AB - BACKGROUND: In the heart, there are high constitutive levels of the two related small heat shock proteins, HSP25 and alphaB-crystallin. To gain insight into their functional role, we have analyzed abundance and location of both proteins in rat and human hearts at different stages of development and in diseased state. METHODS AND RESULTS: Immunoblotting analysis of rat ventricular tissue at fetal, neonatal, and adult stages reveals the level of HSP25 to decline strongly during development, whereas the level of alphaB-crystallin remains nearly constant. In parallel, the portion of phosphorylated isoforms of HSP25 decreases as shown by two-dimensional polyacrylamide gel electrophoresis. HSP25 is detected in cardiomyocytes and endothelial and vascular smooth muscle cells, whereas alphaB crystallin is detected in cardiomyocytes only by immunofluorescence and immunoelectron microscopy. Both proteins colocalize in the I-band and M-line region of myofibrils in cardiomyocytes. In diseased and transplanted adult human hearts, HSP25 and alphaB-crystallin levels are considerably elevated compared with fetal hearts. In failing adult human hearts, phosphorylated isoforms of HSP25 predominate, and cardiomyocytes with a partial dislocation of HSP25 and alphaB-crystallin are observed. CONCLUSIONS: Differential accumulation and location of HSP25 and alphaB-crystallin in heart tissue during development imply distinct functions of both proteins, which seem to be involved in organization of cytoskeletal structures. As judged by level, phosphorylation state, and location of both small heat shock proteins, diseased adult human hearts share features with fetal hearts. PMID- 9396444 TI - Progressive anterior ablation in the coronary sinus region: evidence to support the presence of a 'slow pathway' input in normal patients? AB - BACKGROUND: AV node modification is an emerging approach to rate control in patients with medically refractory atrial fibrillation. The mechanism of benefit of this procedure is not completely understood. METHODS AND RESULTS: Twenty-two patients (age, 65+/-11 years; 16 women) with medically refractory paroxysmal atrial fibrillation referred for complete AV node ablation underwent serial ablations beginning at the level of the coronary sinus os progressing in a superior and anterior direction toward the His bundle. Serial atrial extrastimulus testing was performed to determine the effect of the progressive posteroseptal ablation in the region of the coronary sinus on the AV node antegrade refractory curve. Two of 22 patients had antegrade dual AV node pathways before ablation. Three patterns of response to serial ablation were noted. In 10 patients (45%), loss of the terminal portion of the AV node antegrade refractory curve occurred without evidence of fast pathway injury. In 7 patients (32%) the curve was shifted upward and to the left, consistent with nonspecific AV node damage. In 5 patients (23%), no effect could be attained before induction of complete AV block at superior and anterior ablation sites. Clinical variables and site of ablation did not predict response to serial ablations. CONCLUSIONS: These data suggest that the mechanism of benefit of AV node modification in this population may be through elimination of "slow pathway" tissue in half of patients and nonspecific injury in the remainder. Modification without complete AV block may not be possible in a minority of patients, as the response to progressive ablation appears to be "all or none" conduction. PMID- 9396445 TI - Relationship between atrial fibrillation and typical atrial flutter in humans: activation sequence changes during spontaneous conversion. AB - BACKGROUND: A transitional rhythm precedes the spontaneous onset of atrial flutter in an animal model, but few data are available in man. METHODS AND RESULTS: In 10 patients, 16 episodes of atrial fibrillation (166+/-236 seconds) converting into atrial flutter during electrophysiological evaluation were analyzed. A 20-pole catheter was used for mapping the right atrial free wall. Preceding the conversion was a characteristic sequence of events: (1) a gradual increase in atrial fibrillation cycle length (150+/-25 ms after onset, 166+/-28 ms before conversion, P<.01); (2) an electrically silent period (267+/-45 ms); (3) "organized atrial fibrillation" (cycle length, 184+/-24 ms) with the same right atrial free wall activation direction as during atrial flutter; (4) another delay on the lateral right atrium (283+/-52 ms); and (5) typical atrial flutter (cycle length, 245+/-38 ms). The coronary sinus generally had a different rate than the right atrial free wall until the beat that initiated flutter, when right atrium and coronary sinus were activated in sequence. During 1313 seconds of fibrillation, there were 171 episodes of "organized atrial fibrillation." An additional activation delay at least 30 ms longer than the mean organized atrial fibrillation cycle length was sensitive (100%) and specific (99%) for impending organization into atrial flutter. During organized atrial fibrillation, right atrial free wall activation was craniocaudal in 70% and caudocranial in 30%, which may explain why counterclockwise flutter is a more common clinical rhythm than clockwise flutter. Atrial flutter never degenerated into fibrillation, even after adenosine infusion. CONCLUSIONS: Anatomic barriers, along with statistical properties of conduction and refractoriness during atrial fibrillation, may explain the remarkably stereotypical pattern of endocardial activation during the initiation of atrial flutter via fibrillation and the rarity of degeneration of flutter to fibrillation once it stabilizes. PMID- 9396446 TI - Absence of parasympathetic control of heart rate after human orthotopic cardiac transplantation. AB - BACKGROUND: Partial reinnervation of cardiac sympathetic nerves has been observed after heart transplantation; we hypothesized that parasympathetic control to the heart after transplantation may return as well. To test this hypothesis, we examined heart rate responses produced by two cardiovascular reflexes whose efferent limbs are subserved by vagal fibers to the heart: (1) trigeminal reflex (simulated diving reflex) and (2) arterial baroreflex with phenylephrine injection. METHODS AND RESULTS: An "early" group (n=31, <24 months after transplantation) and a "late group" (n=27, >45 months after transplantation) were studied and compared with a control group with intact cardiac innervation (n=32) and a renal transplant group with similar transplant immunosuppressive regimen (n=11). For trigeminal reflex testing, responses of the donor sinus node (DSN) (sinus node controlling heart rate) and recipient sinus node (RSN) in the innervated remnant right atrium in cardiac transplant patients were compared with heart rate responses in the control groups. For arterial baroreflex testing, baroreflex gains for the DSN and RSN in the cardiac transplant groups were compared with those of the control group. With engagement of the trigeminal reflex, the DSN rate of both transplant groups changed minimally (early, 1.2+/ 1.2 bpm; late, 1.8+/-2.5 bpm) compared with the expected decrease in control subjects (-19.8+/-3.0 bpm) and renal transplant patients (-23.9+/-4.9 bpm) (P<.001 versus cardiac transplants). Changes in the RSN rate of both cardiac transplant groups (early, -13.0+/-4.0 bpm; late, -10.0+/-3.7 bpm) were similar to the control groups. Arterial baroreflex gains for the DSN were also depressed (early, 0.1+/-0.2 ms/mm Hg; late, 0.2+/-0.2 ms/mm Hg) compared with control (14.9+/-1.8 ms/mm Hg) and RSN (early, 9.9+/-1.3 ms/mm Hg; late, 10.9+/-1.3 ms/mm Hg; P<.001 versus DSN transplant). CONCLUSIONS: These data suggest that parasympathetic influences on donor heart rate are absent in the majority of patients up to 96 months after cardiac transplantation. PMID- 9396447 TI - Radiofrequency catheter ablation of postinfarction ventricular tachycardia: long term success and the significance of inducible nonclinical arrhythmias. AB - BACKGROUND: Radiofrequency (RF) catheter ablation is effective therapy for monomorphic ventricular tachycardia (VT) in patients without structural heart disease. In patients with postinfarction VT; however, this procedure has been used predominantly as adjunctive therapy, targeting only the patient's clinically documented arrhythmia. By targeting all inducible, sustained VT morphologies, we sought to determine the utility of RF catheter ablation as a primary cure in patients who present with hemodynamically tolerated VT. METHODS AND RESULTS: RF ablation was attempted in 35 patients with a previous myocardial infarction and recurrent, hemodynamically tolerated VT. A mean of 3.9+/-2.7 VTs were induced per patient (range, 1 to 10). The clinically documented arrhythmia was successfully ablated in 30 of 35 patients (86%), and on follow-up electrophysiological testing, 11 patients had no inducible VT and were discharged without other therapy. Nineteen patients had inducible "nonclinical" arrhythmias on follow-up testing, and the majority underwent cardiac defibrillator implantation. Freedom from recurrent arrhythmias, including sudden death, was 91% in patients without inducible VT and 53% in patients with persistently inducible "nonclinical" arrhythmias (P<.05; mean follow-up, 17+/-12 and 12+/-11 months, respectively). CONCLUSIONS: In patients with well-tolerated VT, RF catheter ablation may be useful as a primary cure if no other ventricular arrhythmias are inducible on follow-up testing. Ablation of all hemodynamically tolerated arrhythmias should be attempted in patients with multiple inducible VT morphologies because of the high rate of recurrence of unablated VTs in these patients. PMID- 9396448 TI - Analysis of the degree of QRS fusion necessary for its visual detection: importance for the recognition of transient entrainment. AB - BACKGROUND: Fixed fusion is the hallmark for the demonstration of transient entrainment. However, the degree of accuracy of its recognition on the surface ECG is unknown. The purpose of the present study was to evaluate the ability to detect fusion in the QRS complex. METHODS AND RESULTS: While pacing the ventricles at a fixed rate, a model of ventricular fusion was created by introducing late extra stimuli at a second site. In this model, the presence and degree of fusion are known. Pacing sites were the RV apex, outflow tract, and left ventricle in various configurations. We analyzed 433 QRS complexes with different degrees of fusion (or no fusion) in 21 patients. Each QRS was "read" by three investigators blinded to intracardiac recordings but having a reference QRS with no fusion. There was a statistically significant correlation between the degree of fusion and its recognition. Fusion was detected with a sensitivity of 75% and a specificity of 87%. Fusion was accurately detected in all configurations only when >22% of the QRS was fused. In patients with organic left ventricular disease, fusion was better recognized when the driving pacing site was the left ventricle than when it was a right ventricular site. The interobserver agreement was moderate between two pairs of observers and only fair between the remaining pair. CONCLUSIONS: Our results suggest that an accurate detection of ventricular fusion can only be accomplished when fusion occurs during a significant proportion of the QRS duration. The potential lack of recognition of minor degrees of fusion may produce underdetection of transient entrainment. PMID- 9396449 TI - Strength-duration relationship for human transvenous defibrillation. AB - BACKGROUND: One of the basic characteristics of electrical defibrillation is the strength-duration relationship, or the effect of pulse width on defibrillation efficacy. This relationship is important for understanding the mechanism of defibrillation and for the design of optimal waveforms. However, a detailed evaluation of the strength-duration relationship for human transvenous defibrillation has not been performed previously. METHODS AND RESULTS: This was a prospective study of 29 patients undergoing initial defibrillator implantation with a uniform dual coil, transvenous lead. In each patient defibrillation thresholds were measured for either short (2, 3, 4, 6 ms) or long (6, 12, 18 ms) pulse durations, with the order of testing randomized. The shock waveform was a truncated monophasic pulse from a capacitor of 150 microF. The leading edge voltage at defibrillation threshold was 566+/-100 V for 2-ms pulses. Voltages declined exponentially with increasing pulse width reaching an asymptote by 6 ms (451+/-68 V, P<.05). Defibrillation threshold voltage was insensitive to longer pulse widths. Stored energy at defibrillation threshold showed a similar relationship with pulse width. In contrast, mean current decreased monotonically over the full range of pulse durations evaluated, and there was no evidence of a rheobase. CONCLUSIONS: The shape of the strength-duration curve and the lack of rheobase current indicate a fundamental difference between cardiac stimulation and defibrillation. The relationship between pulse duration and defibrillation threshold voltage or stored energy is well modeled by a parallel capacitor resistor circuit with a time constant of 5.3 ms. PMID- 9396450 TI - Instant power spectrum analysis of heart rate variability during orthostatic tilt using a time-/frequency-domain method. AB - BACKGROUND: Spectral analysis of heart rate (HR) variability (HRV) requires, as a rule, some level of stationarity and, as a result, is inadequate to quantify biological transients. A time-/frequency-domain method (TF) was developed to obtain an instant spectral power (SP) of HRV during tilt. METHODS AND RESULTS: HR was recorded by Holter monitoring in volunteers and analyzed with a TF, the smoothed pseudo-Wigner-Ville transformation (SPWVT), with the table inclination randomly set or continuously increased while the table rotated in head-up position. (1) The SPWVT assesses, beat by beat, the instant center frequency (ICF) of the SP. ICF correlates better with instant HR than the ratio of low- (LF) to high-frequency (HF) oscillations. The transient effect of tilt is better characterized as a shift of SP toward lower frequencies than by changes in amplitudes. (2) The method evidences variations of HR from one second to another. During the passage to head-up position, the vagal withdrawal and the sympathetic activation occur nearly simultaneously, as indicated by the instant changes in both LF and HF amplitudes and ICF. (3) The averaged results of the SPWVT give results similar to those previously obtained with autoregressive algorithms. CONCLUSIONS: The SPWVT is a new tool to explore HR transitions such as periods before episodes of arrhythmias on a time scale of one beat and allows quantification of an instant frequency index (ICF) that closely reflects the instantaneous relationship between sympathetic and vagal modulations. PMID- 9396451 TI - Evaluation of the specificity of morphological electrocardiographic criteria for the differential diagnosis of wide QRS complex tachycardia in patients with intraventricular conduction defects. AB - BACKGROUND: Although several ECG criteria have been described for the differential diagnosis of tachycardias with a wide QRS complex, their applicability in patients with preexisting intraventricular conduction defects (IVCDs) has been questioned. The specificity of previously described criteria in this context is unknown. METHODS AND RESULTS: We analyzed prospectively the specificity of the QRS morphological criteria previously described in ECGs during sinus rhythm of 232 patients with IVCD. Only 5 of 12 analyzed criteria had a specificity > or = 0.90 among our patients: (1) a triphasic configuration (Rsr' or Rr') QRS complex in V1 in the presence of a right bundle-branch block morphology (BBBM); (2) a QS, QR, or R QRS pattern in V6 in the presence of a right BBBM; (3) any Q in V6 in the presence of a left BBBM; (4) a concordant pattern in all precordial leads; and (5) the absence of an RS complex in all precordial leads (particularly useful for left BBBM). The following criteria--QRS duration > 140 ms; a left axis with right BBBM, right superior axis with right BBBM, monophasic or biphasic R wave in V1 with right BBBM, and a relation R/S < 1 with right BBBM; an R > 30 ms in lead V1 or V2 with left BBBM, > 60 ms from QRS onset to S nadir with left BBBM, a notched downstroke S wave with left BBBM, and an R-to-S interval > 100 ms in one precordial lead--had a specificity of 0.43, 0.54, 0.87, 0.80, 0.85, 0.78, 0.66, 0.69, and 0.63 (0.84 in right BBBM), respectively. CONCLUSIONS: Most of the previously described morphological criteria favoring ventricular tachycardia are present in a substantial percentage of patients with IVCD during sinus rhythm. These findings suggest a limited applicability of these criteria in this subset of patients. PMID- 9396452 TI - Cell-derived microparticles generated in patients during cardiopulmonary bypass are highly procoagulant. AB - BACKGROUND: Microparticles from platelets and other cells have been extensively studied and characterized in vitro. Although the level of platelet-derived microparticles is elevated in a variety of diseases, including cardiac surgery, virtually nothing is known about their functions in vivo. The aim of the present study was to investigate the procoagulant properties of microparticles generated in vivo. METHODS AND RESULTS: In 6 patients at the end of cardiopulmonary bypass, 14.8 x 10(9)/L (median; range, 9.7 to 27.4 x 10(9)/L) platelet-derived microparticles were present in pericardial blood, whereas blood obtained from the systemic circulation contained 1.6 x 10(9)/L (median; range, 0.4 to 8.9 x 10(9)/L) of such microparticles, as determined by flow cytometry. Microparticles stained positively for phosphatidylserine as determined with labeled annexin V. In contrast to systemic blood, pericardial blood contained not only microparticles of platelet origin but also microparticles that originated from erythrocytes, monocytes, or granulocytes, and other hitherto unknown cellular sources. Plasma prepared from pericardial blood and to a lesser extent plasma from systemic blood obtained at the same time, stimulated formation of thrombin in vitro. This activity of pericardial plasma was lost after removal of its microparticles by high-speed centrifugation, whereas the corresponding microparticle pellet was strongly procoagulant. The generation of thrombin in vitro involved a tissue factor/factor VII-dependent and factor XII-independent pathway. CONCLUSIONS: This study is the first to demonstrate that microparticles generated in vivo can stimulate coagulation. PMID- 9396453 TI - Activation of the complement system during and after cardiopulmonary bypass surgery: postsurgery activation involves C-reactive protein and is associated with postoperative arrhythmia. AB - BACKGROUND: Complement activation during cardiopulmonary bypass (CPB) surgery is considered to result from interaction of blood with the extracorporeal circuit. We investigated whether additional mechanisms may contribute to complement activation during and after CPB and, in particular, focused on a possible role of the acute-phase protein C-reactive protein (CRP). METHODS AND RESULTS: In 19 patients enrolled for myocardial revascularization, perioperative and postoperative levels of complement activation products, interleukin-6 (IL-6), CRP, and complement-CRP complexes, reflecting CRP-mediated complement activation in vivo, were measured and related to clinical symptoms. A biphasic activation of complement was observed. The ratio between the areas under the curve of perioperative and postoperative C3b/c and C4b/c were 3:2 and 1:46, respectively. IL-6 levels reached a maximum at 6 hours post-surgery. CRP levels peaked on the second postoperative day. Each complement-CRP complex had peak levels on the second or third postoperative day. By multivariate analysis, maximum levels of CRP on the second postoperative day were mainly explained by C4b/c levels after protamine administration, leukocyte count on the second postoperative day, and preoperative levels of CRP. Peak levels of C4b/c after protamine administration (P=.0073) and on the second postoperative day correlated with the occurrence of arrhythmia on the same day (P=.0065). CONCLUSIONS: Cardiac surgery with CPB causes a biphasic complement activation. The first phase occurs during CPB and results from the interaction of blood with the extracorporeal circuit. The second phase, which occurs during the first 5 days after surgery, involves CRP, is related to baseline CRP levels, and is associated with clinical symptoms such as arrhythmia. PMID- 9396454 TI - Association of parvovirus B19 genome in children with myocarditis and cardiac allograft rejection: diagnosis using the polymerase chain reaction. AB - BACKGROUND: Inflammatory diseases of the heart, including myocarditis and cardiac transplant rejection, are important causes of morbidity and mortality in children. Although viral infection may be suspected in either of these clinical conditions, the definitive etiology is often difficult to ascertain. Furthermore, the histology is identical for both disorders. Coxsackievirus has long been considered the most common cause of viral myocarditis; however, we previously demonstrated by polymerase chain reaction (PCR) analysis that many different, and sometimes unexpected, viruses may be responsible for myocarditis and cardiac rejection. In this study, we describe the association of parvovirus genome identified through PCR analysis of cardiac tissue in the clinical setting of myocarditis and cardiac allograft rejection. METHODS AND RESULTS: Myocardial tissue from endomyocardial biopsy, explant, or autopsy was analyzed for parvovirus B19 using primers designed to amplify a 699-base pair PCR product from the VP1 gene region. Samples tested included those obtained from patients with suspected myocarditis (n=360) or transplant rejection (n=200) or control subjects (n=250). Parvoviral genome was identified through PCR in 9 patients (3 myocarditis; 6 transplant) and no control patients. Of the 3 patients with myocarditis, 1 presented with cardiac arrest leading to death, 1 developed dilated cardiomyopathy, and the other gradually improved. Four of the 6 transplant patients had evidence of significant rejection on the basis of endomyocardial biopsy histology. All transplant patients survived the infection. CONCLUSIONS: Parvovirus is associated with myocarditis in a small percentage of children and may be a potential contributor to cardiac transplant rejection. PCR may provide a rapid and sensitive method of diagnosis. PMID- 9396455 TI - Primate smooth muscle cell migration from aortic explants is mediated by endogenous platelet-derived growth factor and basic fibroblast growth factor acting through matrix metalloproteinases 2 and 9. AB - BACKGROUND: Migration of arterial smooth muscle cells (SMCs) is regulated by basic fibroblast growth factor (bFGF), platelet-derived growth factor (PDGF), and matrix metalloproteinases (MMPs) in the injured rat carotid artery. We have recently shown that migration of SMCs from baboon aortic explants depends on the activity of MMPs, but the identity of the stimulatory MMPs and the role of bFGF and PDGF in this primate system are not known. METHODS AND RESULTS: These experiments were designed to determine whether MMP2, MMP9, bFGF, or PDGF plays a role in SMC migration from medial explants of baboon aorta. Explants were cultured in serum-free medium with insulin, transferrin, and ovalbumin. Neutralizing antibodies to MMP2 and antibodies that inhibit activation of proMMP9 decreased SMC migration from the aortic explants. Antibodies to bFGF and to the alpha- and beta-subunits of the PDGF receptor also inhibited migration from the explants. Addition of bFGF and PDGF-BB but not PDGF-AA increased migration. The antibodies to bFGF but not the antibodies to the PDGF receptor subunits decreased the levels of MMP9, whereas all the antibodies decreased activated MMP2. CONCLUSIONS: These data demonstrate that SMC migration from primate aortic explants is dependent on endogenous MMP2, MMP9, PDGF, and bFGF. The data also suggest that PDGF-induced (PDGF-BB or possibly PDGF-AB) migration is dependent on MMP2, whereas bFGF-induced migration depends on both MMP2 and MMP9. PMID- 9396456 TI - In utero cardiac gene transfer via intraplacental delivery of recombinant adenovirus. AB - BACKGROUND: The relationship among the maternal, placental, and uniquely shunted embryonic circulation was explored to provide access to the embryonic cardiovascular system in utero. Manipulation of gene expression in the developing heart would be particularly useful for studying the effects of altered gene expression on cardiac development and in the etiology of congenital cardiac anomalies. METHODS AND RESULTS: Dye studies demonstrated that intraplacental injection allows direct access to the embryonic cardiac and systemic circulation. To evaluate the efficacy of cardiac gene transfer using this approach, replication-deficient recombinant adenoviral vectors encoding luciferase or beta galactosidase as reporter genes were injected intraplacentally into embryonic day (E)12.5 murine embryos, an age at which the mass of the heart was observed to be large compared with other organs. Embryos were assayed for transgene expression at E15.5 and at birth. Survival rates at these times were similar among vector injected and control groups. At E15.5 and at birth, luciferase activity within the heart was 9- and 23-fold higher, respectively, than in the remainder of the embryo, although levels of expression were generally lower at birth than during embryonic life. Beta-galactosidase expression was observed within all regions of the embryonic heart and was localized to approximately 15% of atrial and ventricular cells. CONCLUSIONS: Intraplacental delivery of adenovirus at embryonic day 12.5 results in somatic gene transfer to the murine embryonic heart, which persists at least until birth. The combination of intraplacental injection to directly access the fetal coronary circulation and injection at E12.5 when the mass of the heart is large compared with other organs results in transgene expression in cardiac cells. Intraplacental injections early in embryonic life may thus be useful to study the effects of temporal manipulation of gene expression on cardiac development and disease. PMID- 9396457 TI - Cardiovascular phenotype of a mouse strain with disruption of bradykinin B2 receptor gene. AB - BACKGROUND: To evaluate the role of kinins in the regulation of cardiovascular function, we studied the phenotype of a mouse strain with disruption of the bradykinin B2-receptor gene (Bk 2r-/-). METHODS AND RESULTS: Under basal conditions, tail-cuff blood pressure was higher in Bk2r-/- than in wild-type Bk2r+/+ and heterozygous Bk2r+/- mice (124+/-1 versus 109+/-1 and 111+/-2 mm Hg, respectively; P<.01 for both comparisons), a difference that was confirmed by measurements of intra-arterial blood pressure in unanesthetized mice. Heart weight was greater in Bk2r-/- than in Bk2r+/+ and Bk2r+/- mice (505+/-10 versus 449+/-12 and 477+/-10 mg/100 g body wt, P<.05). Chronic blockade of B2-receptors by Icatibant (50 nmol/100 g body wt twice a day S.C.) or inhibition of nitric oxide synthase by nitro-L-arginine-methyl ester (0.14 mmol/100 g body wt orally) increased the blood pressure of Bk2r+/+ to the levels of Bk2r-/- mice. Compared with the wild-type strain, both Bk2r-/- and Bk2r+/- mice showed exaggerated vasopressor responses to angiotensin II. In addition, chronic administration of an angiotensin AT1-receptor antagonist reduced the basal blood pressure of Bk2r-/ by 21+/-3 mmHg (P<.05) to the levels of Bk2r+/+. No difference was detected between strains as far as plasma renin activity and the expression of renin and AT1-receptor genes are concerned. Chronic salt loading (0.84 mmol/g chow for 15 days) increased the blood pressure of Bk2r-/- and Bk2r+/- by 34+/-3 and 14+/-6 mm Hg, respectively, whereas it was ineffective in Bk2r+/+. CONCLUSIONS: Our results suggest that a normally functioning B2-receptor is essential for the maintenance of cardiovascular homeostasis in mice. Dysfunction of the kallikrein-kinin system could contribute to increase blood pressure levels by leaving the activity of vasoconstrictor agents unbalanced. PMID- 9396459 TI - Intimal hyperplasia after balloon injury is attenuated by blocking selectins. AB - BACKGROUND: Cell adhesion molecules facilitate the adherence of platelets and leukocytes to the vascular endothelium in response to injury. Restenosis after balloon angioplasty is thought to represent the response to vascular injury. The role of cell adhesion in this process is unclear. METHODS AND RESULTS: This study was performed in New Zealand White rabbits that underwent balloon angioplasty of the iliac artery. Expression of the cell adhesion molecule E-selectin on endothelium was determined by immunohistochemistry and increased at 6 hours with a peak expression 24 to 48 hours after balloon injury, returning to baseline by 1 week. The expression of L-selectin on circulating leukocytes, measured by flow cytometry, was significantly increased at 48 hours, with return to baseline by 1 week. In seven animals, the selectins were blocked with an analogue of sialyl Lewis(x) given as an I.V. bolus of 10 mg/kg followed by 2 mg x kg(-1) x h(-1) I.P. infusion for 7 days. After 4 weeks, compared with control animals, the study group had a larger lumen area (57.7 versus 44.7 mm2, P<.05), smaller intima area (9.0 versus 19.2 mm2, P<.01), smaller intima/media ratio (0.4 versus 1.0, P<.01), and a smaller percent area stenosis (15.6% versus 34.3%, P<.01). CONCLUSIONS: The cell adhesion molecules E-selectin and L-selectin are expressed after balloon injury. Blockade of the selectins has a favorable effect on the response to vascular injury. PMID- 9396458 TI - Gap junction uncoupler heptanol prevents cell-to-cell progression of hypercontracture and limits necrosis during myocardial reperfusion. AB - BACKGROUND: The objective of this study was to test the hypothesis that chemical interaction through gap junctions may result in cell-to-cell progression of hypercontracture and that this phenomenon contributes to the final extent of reperfused infarcts. METHODS AND RESULTS: Cell-to-cell transmission of hypercontracture was studied in pairs of freshly isolated adult rat cardiomyocytes. Hypercontracture induced by microinjection of a solution containing 1 mmol/L Ca2+ and 2% lucifer yellow (LY) was transmitted to the adjacent cell (11 of 11 pairs), and the gap junction uncoupler heptanol (2 mmol/L) prevented transmission in 6 of 8 pairs (P=.003), with a perfect association between passage of the LY and transmission of hypercontracture. In the isolated, perfused rat heart submitted to 30 minutes of hypoxia, addition of heptanol to the perfusion media during the first 15 minutes of reoxygenation had a dose-related protective effect against the oxygen paradox, as demonstrated by a reduction of diastolic pressure and marked recovery of developed pressure (P<.001), as well as less lactate dehydrogenase release during reoxygenation (P<.001) and less contraction band necrosis (P<.001) than controls. In the in situ pig heart submitted to 48 minutes of coronary occlusion, the intracoronary infusion of heptanol during the first 15 minutes of reperfusion at a final concentration of 1 mmol/L limited myocardial shrinkage, reflecting hypercontracture (P<.05), reduced infarct size after 5 hours of reperfusion by 54% (P=.04), and modified infarct geometry with a characteristic fragmentation of the area of necrosis. Heptanol at 1 mmol/L had no significant effect on contractility of nonischemic myocardium. CONCLUSIONS: These results demonstrate that hypercontracture may be transmitted to adjacent myocytes through gap junctions and that heptanol may interfere with this transmission and reduce the final extent of myocardial necrosis during reoxygenation or reperfusion. These findings are consistent with the hypothesis tested and open a new approach to limitation of infarct size by pharmacological control of gap junction conductance. PMID- 9396460 TI - Vitamins C and E inhibit O2- production in the pig coronary artery. AB - BACKGROUND: We previously found in a pig coronary balloon injury model that vitamins C and E as well as probucol had beneficial effects on the vessel response to injury measured by morphometry These effects correlated with an inhibition in the ability to oxidize LDLs ex vivo, suggesting that the morphological response was due to the antioxidant effect of the treatments. METHODS AND RESULTS: In the present study, the production of O2- by vessels 14 days after balloon injury was determined and correlated with circulating and tissue levels of vitamins C and E. Twenty-five domestic pigs were divided into four groups: control (n=7), vitamin C (500 mg/d, group C, n=6), vitamin E (1000 IU/d, group E, n=6), and vitamins C and E (500 mg/d and 1000 IU/d, group C+E, n=6). Vitamins were administered 7 days before oversized balloon injury of the left anterior descending coronary artery (LAD) and continued for 14 days after injury. Vitamin C and E concentrations were determined in plasma and lymphocytes as an index for tissue levels. Vessels were harvested after animals were killed, and O2- production was measured by lucigenin chemiluminescence. O2- production by the injured LAD was 2.5-fold greater than O2- production by the uninjured LAD or right coronary artery (RCA). The increase in O2- was caused primarily by cells present in the media and neointima. All vitamin-treated groups showed significantly decreased O2- production in both the RCA and LAD (approximately 45% inhibition) relative to vessels in the control, untreated group. There was a significant correlation between LAD O2- production and lymphocyte vitamin E levels. CONCLUSIONS: The present study is the first to show increased O2- production in injured vessels and to demonstrate that antioxidant vitamins reduce O2- production. These results suggest that beneficial effects of antioxidant vitamins in coronary artery disease are related, in part, to alterations in vessel redox state. PMID- 9396461 TI - Differential effect of hydrogen peroxide and superoxide anion on apoptosis and proliferation of vascular smooth muscle cells. AB - BACKGROUND: Proliferation and apoptosis of vascular smooth muscle cells (VSMCs) are two important components of atherosclerosis, restenosis, and hypertension. Although reactive oxygen species have been demonstrated to participate in the pathogenesis of these diseases, their precise involvement has not been fully understood. We hypothesized that different reactive oxygen species exert distinct effects on proliferation and apoptosis of VSMCs. METHODS AND RESULTS: Cultured rat VSMCs were exposed to xanthine oxidase/xanthine (XO/X) or H2O2-Fe(II). A single exposure to XO/X predominantly resulted in cell proliferation, whereas frequent exposures to high levels of XO/X predominantly resulted in cell death. Administration of superoxide dismutase and catalase revealed that O2- but not H2O2, was mitogenic to VSMCs, whereas H2O2 was responsible for VSMC death. Treatment with H2O2-Fe(II) alone or in the presence of different hydroxyl radical scavengers showed that VSMC death occurred in a dose-dependent manner and was mediated by the formation of hydroxyl radicals. Cell death caused by XO/X or H2O2 Fe(II) occurred by apoptosis as revealed by condensation of nuclei, appearance of a "DNA ladder," increases in DNA fragmentation, and positive in situ nick-end labeling. Northern blot analysis indicated that bcl-2 and c-fos but not p53 and c myc may participate in mediating H2O2-Fe(II)-induced VSMC apoptosis. CONCLUSIONS: Different reactive oxygen species exert distinct effects on VSMCs, with O2- inducing proliferation and H2O2 causing apoptosis. Thus, reactive oxygen species might participate in atherosclerosis, restenosis, and hypertension in a dual manner by stimulating proliferation and triggering apoptosis of VSMCs. PMID- 9396462 TI - Pulsatile stretch stimulates superoxide production in human aortic endothelial cells. AB - BACKGROUND: Free radicals such as superoxide and nitric oxide (NO) play a key role in the pathophysiology of atherosclerosis. Mechanical forces such as pulsatile stretch may be involved in free radical production. We studied superoxide production by pulsatile stretch in human endothelial cells. METHODS AND RESULTS: Human cultured aortic endothelial cells were exposed to pulsatile stretch up to 24 hours, and superoxide production was examined. Short-term stretch for 1 hour (10% average elongation, 50 cycles per minute) increased superoxide production 2.2-fold. This effect was reduced by diphenyleneiodonium chloride, an NADPH oxidase inhibitor, but not by the xanthine oxidase inhibitor oxypurinol or the cyclooxygenase inhibitor indomethacin. Prolonged stretch up to 6 hours increased superoxide production, but it returned to near the control level after 24 hours of stretch. However, after blockade of NO production, 24 hours of stretch did increase superoxide production 2.4-fold compared with 24 hours of stretch alone. Moreover, 24-hour stretch doubled NO synthase (NOS) (III) protein and mRNA expression. The tetrahydrobiopterin synthesis inhibitor 2,4 diamino-6-hydroxypyrimidine had no effect on unstretched cells but doubled superoxide production compared with 24-hour stretch alone; this increase was halved by cotreatment with 6-methyl-5,6,7,8-tetrahydropterine, a lipid-soluble form of tetrahydrobiopterine. CONCLUSIONS: Short-term stretch increased superoxide production from human aortic endothelial cells via NADPH oxidase and NOS (III), whereas prolonged stretch increased both superoxide and NO production. The increase in NOS (III) protein with prolonged stretch acts as a scavenger mechanism whereby NO inactivates superoxide. Tetrahydrobiopterin determines the balance of superoxide and NO production from NOS (III) after prolonged stretch in which NOS (III) level is upregulated. PMID- 9396463 TI - Na+/H+ exchanger activity does not contribute to protection by ischemic preconditioning in the isolated rat heart. AB - BACKGROUND: Despite evidence that pharmacological inhibition of the Na+/H+ exchanger (NHE) is cardioprotective, activation of NHE has been proposed as a protective mechanism of ischemic preconditioning (PC). METHODS AND RESULTS: In isolated rat ventricular myocytes (n=8 to 11 per group) loaded with the fluorescent pH indicator C-SNARF-1, we showed that HOE-642 (HOE) was a potent inhibitor of the sarcolemmal NHE (80% inhibition at 1 micromol/L); such inhibition was readily reversible by washout of the drug. We confirmed that 1 micromol/L HOE produces significant and reversible inhibition of NHE activity in isolated rat hearts as well (n=4), and in this model, we tested (n=8 per group) whether the presence of the drug during (1) the prolonged period of ischemia (40 or 60 minutes) or (2) the preceding brief periods of PC ischemia (3 minutes plus 5 minutes) modulates the protective efficacy of PC. In protocol 1, HOE was infused for 5 minutes immediately before the prolonged ischemic period. With 40 minutes of prolonged ischemia, the postischemic recovery of left ventricular developed pressure (LVDP) was 15+/-2% in controls and was improved to 45+/-7% with HOE (P<.05), 55+/-5% with PC (P<.05), and 68+/-2% with PC+HOE (P<.05 versus all groups). When the prolonged ischemic period was extended to 60 minutes, an additive effect of PC and HOE was readily apparent and LVDP recovery with PC+HOE (66+/-2%) was almost double that observed with HOE (37+/-4%) or PC (34+/-5%) alone (P<.05). In protocol 2, HOE was infused for 3 minutes immediately before each episode of PC ischemia and was subsequently washed out before a 40-minute prolonged ischemic period (HOE+PC). LVDP recovery was 34+/-4% in controls and was improved to 57+/-2% with PC (P<.05) and 55+/-3% with HOE+PC (P<.05). Improved recovery of LVDP was matched by reduced creatine kinase leakage in all cases. CONCLUSIONS: Because coadministration of HOE (at a concentration sufficient to inhibit NHE activity) did not reduce the efficacy of PC in either protocol, we conclude that NHE activity does not contribute to the cardioprotective actions of PC. On the contrary, NHE inhibition during the prolonged ischemic period may enhance the protection afforded by PC. PMID- 9396464 TI - Physiological concentrations of estradiol attenuate endothelin 1-induced coronary vasoconstriction in vivo. AB - BACKGROUND: Estrogens are cardioprotective hormones and are reported to have antianginal properties. We examined the effect of physiological concentrations of 17beta-estradiol on coronary reactivity in anesthetized female farm pigs. METHODS AND RESULTS: Epicardial coronary cross-sectional area (CSA) was assessed by two dimensional intravascular ultrasound, average coronary peak flow velocity (APV) by intravascular Doppler velocimetry, and coronary blood flow (CBF) was calculated. Dose-response curves to intracoronary endothelin-1 (ET-1, 1 pmol/L to 10 nmol/L), the selective ET(B) receptor agonist sarafotoxin (1 pmol/L to 10 nmol/L), and serotonin (0.1 nmol/L to 1 micromol/L) were assessed before and after a 10-minute infusion of intracoronary estradiol (1 nmol/L). Before estradiol administration, ET-1 induced significant dose-dependent decreases in CSA, APV, and CBF. Estradiol attenuated ET-1-induced epicardial vasoconstriction (P<.001) as well as ET-1-induced decreases in APV (P=.05) and CBF (P=.012). In an additional five pigs, vehicle (DMSO) had no effect on ET-1-induced coronary vasoconstriction. Before estradiol administration, sarafotoxin induced no net change in CSA but induced increases in APV and CBF, the extent of which did not change significantly after estradiol. Serotonin induced small decreases in CSA but increased APV and CBF. Estradiol did not influence serotonin-induced changes in CSA, APV, or CBF. CONCLUSIONS: We conclude that estradiol attenuates ET-1 induced vasoconstriction, possibly through effects on the ET(A) receptor, because selective ET(B) receptor-induced stimulation with sarafotoxin remained unchanged. Such an effect on the ET(A) receptor may relate to the antianginal properties of estrogens. PMID- 9396465 TI - Human protein C receptor is present primarily on endothelium of large blood vessels: implications for the control of the protein C pathway. AB - BACKGROUND: The protein C anticoagulant pathway is critical to the control of hemostasis. Thrombomodulin and a newly identified receptor for protein C/activated protein C, EPCR, are both present on endothelium. EPCR augments activation of protein C by the thrombin-thrombomodulin complex. METHODS AND RESULTS: To gain a better understanding of the relationship between thrombomodulin and EPCR, we compared the cellular specificity and tissue distributions of these two receptors by using immunohistochemistry. EPCR expression was detected almost exclusively on endothelium in human and baboon tissues. In most organs, EPCR was expressed relatively intensely on the endothelium of all arteries and veins, most arterioles, and some postcapillary venules. EPCR staining was usually negative on capillary endothelial cells. In contrast, thrombomodulin was detected at high concentrations in both large vessels and capillary endothelium. Both thrombomodulin and EPCR were expressed poorly on brain capillaries. The liver sinusoids were the only capillaries in which EPCR was expressed at moderate levels and thrombomodulin was low. EPCR and thrombomodulin were both expressed on the endothelium of vasa recta in the renal medulla, the lymph node subcapsular and medullary sinuses, and some capillaries within the adrenal gland. Even in these organs the majority of capillaries were EPCR negative or stained weakly. CONCLUSIONS: These studies suggest that EPCR may be important in enhancing protein C activation on large vessels. The presence of high levels of EPCR on arterial vessels may help explain why partial protein C deficiency is a weak risk factor for arterial thrombosis. PMID- 9396466 TI - Differential effects of endothelin receptor activation on cyclic flow variations in rat mesenteric arteries. AB - BACKGROUND: Cyclic flow variations (CFVs) represent repetitive cycles of platelet adherence-aggregation and vasoconstriction, followed by dislodgment of platelet thrombi and restoration of blood flow at the site of vascular injury. Although activation of endothelin A (ETA) and endothelin B (ETB) receptors leads to vasoconstriction and nitric oxide release, respectively, the roles of endogenous endothelin-1 (ET-1) and its receptors in CFVs are unknown. METHODS AND RESULTS: A side branch of a mesenteric artery of male Wistar rats was cannulated and a short segment of the artery was mechanically injured to induce CFVs. After 20 minutes of saline infusion, either saline (negative control), BQ-123 (ETA receptor antagonist, 10 microg/min), BQ-788 (ETB receptor antagonist, 10 microg/min), or sarafotoxin S6c (ETB receptor agonist, 10 ng/min) was infused for 20 minutes from the side branch into the injured arterial segment. Percent (%) luminal stenosis as well as proximal and distal vessel diameters were observed and quantitatively measured every minute using intravital video microscopy and a micrometer calibrated video screen. Both BQ-123 and sarafotoxin S6c significantly reduced CFVs represented by the mean luminal stenosis (BQ-123=29+/-13% and sarafotoxin S6c=27+/-11% reduction, respectively; P<.05 for both, compared with saline). In contrast, BQ-788 significantly increased CFVs (33+/-6% increase, P<.05 compared with saline). Moreover, the inhibitory effect of sarafotoxin S6c on CFVs was completely abolished in the presence of N(omega)-nitro-L-arginine methyl ester (L NAME) (a nitric oxide synthase inhibitor, 10(-5) mol/L) in superfusate over the arteries (16.1+/-5% increase, P=NS compared with saline in the presence of L NAME). In addition, BQ-123 caused a significant increase in the diameter of the vessel distal to the injured segment (12+/-4% increase, P<.05 compared with saline). CONCLUSIONS: Endogenous ET-1 release from sites of vascular injury contributes to CFVs and vasomotor tone in the rat mesenteric artery CFV model. ETA and ETB receptors have differential roles in CFVs: ETA receptor antagonism and ETB receptor stimulation reduce CFVs, the latter at least partially through increased nitric oxide formation. PMID- 9396468 TI - Impaired nitric oxide-mediated renal vasodilation in rats with experimental heart failure: role of angiotensin II. AB - BACKGROUND: Congestive heart failure (CHF) is associated with a decrease in renal perfusion. Because endothelium-derived NO is important in the regulation of renal blood flow (RBF), we tested the hypothesis that an impairment in the NO system may contribute to the decrease in RBF in rats with experimental CHF. METHODS AND RESULTS: Studies were performed in rats with experimental high-output CHF induced by aortocaval (AV) fistula and sham-operated controls. In controls, incremental doses of acetylcholine (ACh, 1 to 100 microg x kg(-1) x min(-1)) increased RBF and caused a dose-related decrease in renal vascular resistance (RVR). However, the increase in RBF and decrease in RVR were markedly attenuated in rats with CHF. Likewise, the effects of ACh on urinary sodium and cGMP excretion were also diminished in CHF rats, as was the renal vasodilatory effect of the NO donor S nitroso-N-acetylpenicillamine (SNAP). These attenuated responses to endothelium dependent and -independent renal vasodilators in CHF rats occurred despite a normal baseline and stimulated NO2+NO3 excretion and normal expression of renal endothelial NO synthase (eNOS), as determined by eNOS mRNA levels and immunoreactive protein. Infusion of the NO precursor L-arginine did not affect baseline RBF or the response to ACh in rats with CHF. However, administration of the nonpeptide angiotensin II receptor antagonist A81988 before ACh completely restored the renal vasodilatory response to ACh in CHF rats. CONCLUSIONS: This study demonstrates that despite a significant attenuation in the NO-related renal vasodilatory responses, the integrity of the renal NO system is preserved in rats with chronic AV fistula. This impairment in NO-mediated renal vasodilation in experimental CHF appears to be related to increased activity of the renin angiotensin system and may contribute further to the decrease in renal perfusion seen in CHF. PMID- 9396467 TI - Ca2+ release from intracellular stores is an initial step in hypoxic pulmonary vasoconstriction of rat pulmonary artery resistance vessels. AB - BACKGROUND: A reduction in oxygen tension in the lungs is believed to inhibit a voltage-dependent K+ (Kv) current, which is thought to result in membrane depolarization leading to hypoxic pulmonary vasoconstriction (HPV). However, the direct mechanism by which hypoxia inhibits Kv current is not understood. METHODS AND RESULTS: Experiments were performed on rat pulmonary artery resistance vessels and single smooth muscle cells isolated from these vessels to examine the role of Ca2+ release from intracellular stores in initiating HPV. In contractile experiments, hypoxic challenge of endothelium-denuded rat pulmonary artery resistance vessels caused either a sustained or transient contraction in Ca2+ containing or Ca2+-free solution, respectively (n=44 vessels from 11 animals). When the ring segments were treated with either thapsigargin (5 micromol/L), ryanodine (5 micromol/L), or cyclopiazonic acid (5 micromol/L) in Ca2+-containing or Ca2+-free solution, a significant increase in pulmonary arterial tone was observed (n=44 vessels from 11 animals). Subsequent hypoxic challenge in the presence of each agent produced no further increase in tone (n=44 vessels from 11 animals). In isolated pulmonary resistance artery cells loaded with fura 2, hypoxic challenge, thapsigargin, ryanodine, and cyclopiazonic acid resulted in a significant increase in [Ca2+]i (n=18 cells from 6 animals) and depolarization of the resting membrane potential (n=22 cells from 6 animals). However, with prior application of thapsigargin, ryanodine, or cyclopiazonic acid, a hypoxic challenge produced no further change in [Ca2+]i (n=18 from 6 animals) or membrane potential (n=22 from 6 animals). Finally, application of an anti-Kv1.5 antibody increased [Ca2+]i and caused membrane depolarization. Subsequent hypoxic challenge resulted in a further increase in [Ca2+]i with no effect on membrane potential (n=16 cells from 4 animals). CONCLUSIONS: In rat pulmonary artery resistance vessels, an initial event in HPV is a release of Ca2+ from intracellular stores. This rise in [Ca2+]i causes inhibition of voltage-dependent K+ channels (possibly Kv1.5), membrane depolarization, and an increase in pulmonary artery tone. PMID- 9396469 TI - Stabilization of chronic remodeling by asynchronous cardiomyoplasty in dilated cardiomyopathy: effects of a conditioned muscle wrap. AB - BACKGROUND: Dynamic cardiomyoplasty is a promising new therapy for dilated cardiomyopathy. The girdling effects of a conditioned muscle wrap alone have recently been postulated to partly explain its mechanism. We investigated this effect in a canine model of chronic dilated cardiomyopathy. METHODS AND RESULTS: Twenty dogs underwent rapid ventricular pacing (RVP) for 4 weeks to create a model of dilated cardiomyopathy. Seven dogs were then randomly selected to undergo subsequent cardiomyoplasty, and all dogs had 6 weeks of additional RVP. The cardiomyoplasty group also received 6 weeks of concurrent skeletal muscle stimulation consisting of single twitches delivered asynchronously at 2 Hz to transform the wrap without active assistance. All dogs were studied by pressure volume analysis and echocardiography at baseline and after 4 and 10 weeks of pacing. Systolic indices, including ejection fraction (EF), end-systolic elastance (Ees), and preload-recruitable stroke work (PRSW) were all increased at 10 weeks in the wrap versus controls (EF, 34.0 versus 27.1, P=.008; Ees, 1.65 versus 1.26, P=.09; PRSW, 35.9 versus 25.5, P=.001). Ventricular volumes, diastolic relaxation, and left ventricular end-diastolic pressures stabilized in the cardiomyoplasty group but continued to deteriorate in controls. Both the end systolic and end-diastolic pressure-volume relationships shifted farther rightward in controls but remained stable in the cardiomyoplasty group. CONCLUSIONS: In addition to potential benefits from active systolic assistance, benefits from dynamic cardiomyoplasty appear to be partially accounted for by the presence of a conditioned muscle wrap alone. This conditioned wrap stabilizes the remodeling process of heart failure, arresting progressive deterioration of systolic and diastolic function. PMID- 9396470 TI - Hemodynamic evaluation of the heart with a nonfluoroscopic electromechanical mapping technique. AB - BACKGROUND: Clinical cardiac volumetric measurement techniques are essential for assessing cardiac performance but produce significant inaccuracies in extrapolation of the volume of a three-dimensional (3D) object from two dimensional images and lack the ability to associate cardiac electrical and mechanical activities. In this study, we tested the accuracy of cardiac volumetric measurements using a new catheter-based system. METHODS AND RESULTS: The system uses magnetic technology to accurately locate a special catheter at a frequency of 125 Hz and is currently used in the field of electrophysiology, in which activation maps are superimposed on the 3D geometry of the cardiac chamber. The mapping procedure is based on sequentially acquiring the location of the tip and local electrogram while in contact with the endocardium. The 3D geometry of the chamber is reconstructed in real time, and its volume could be calculated at every time step (8 ms). The volumetric measurements of the system were found to be highly accurate for simple phantoms (mean+/-SEM deviation, 2.3+/-1.1%), left ventricular casts (9.6+/-1.3%), and a dynamic test jig. In addition, left ventricular volumes of 12 swine were measured. Intraobserver and interobserver variabilities were found to be minimal (ejection fraction, 6.5+/-1.9% and 7.1+/ 2.0%; stroke volume, 4.5+/-1.0% and 11.3+/-2.4%). Comparison with the thermodilution method for measuring stroke volume showed an average deviation of 8.1+/-2.2%. Typical pressure-volume loops were also obtained. CONCLUSIONS: The new mapping image provides, for the first time, simultaneous information regarding cardiac mechanics, hemodynamics, and electrical properties. Furthermore, all this information is achieved without the use of fluoroscopy, contrast medium, or complicated image processing. PMID- 9396471 TI - Etomoxir improves left ventricular performance of pressure-overloaded rat heart. AB - BACKGROUND: Numerous studies have demonstrated diverse abnormalities in subcellular structures of pressure-overloaded hypertrophied and failing heart. Long-term administration of etomoxir, a carnitine palmitoyltransferase-1 inhibitor, partially normalized the proportion of myosin isozyme V1 and number of active Ca2+ pumps in hypertrophied rat myocardium. METHODS AND RESULTS: To test the hypothesis that long-term etomoxir treatment improves the performance of hypertrophied ventricle, sham-operated rats and rats with ascending aorta constriction were treated with racemic etomoxir (15 mg/kg per day) for 12 weeks. Left ventricular geometry, dynamics of isovolumic contractions, as well as myosin isozymes as marker of etomoxir-induced phenotype changes were assessed. Etomoxir stimulated (P<.05) slight hypertrophic growth in right and left ventricles of sham-operated rats as well as in right ventricles but not in overloaded left ventricles of rats with aortic constriction. In all treated rats, etomoxir increased (P<.05) maximal developed pressure, left ventricular pressure-volume area, and +/- dP/dt(max). Enhanced values (P<.05) of derived indexes of myocardial performance (normalized stress-length area, maximal rate of wall stress rise, and decline) indicated that myocardial changes were responsible for the improved performance. The etomoxir treatment increased selectively (P<.05) the proportion of myosin V1 in pressure-overloaded left ventricles. CONCLUSIONS: The long-term treatment with etomoxir improved functional capacity of pressure overloaded left ventricle, which can be attributed to an enhanced myocardial performance. Chronic carnitine palmitoyltransferase-1 inhibition may thus represent a candidate approach for developing novel agents that are useful in the prevention of undesirable consequences of pressure overload-induced cardiac hypertrophy. PMID- 9396472 TI - Direct measurement of three-dimensionally reconstructed flow convergence surface area and regurgitant flow in aortic regurgitation: in vitro and chronic animal model studies. AB - BACKGROUND: Evaluation of flow convergence (FC) with two-dimensional (2D) imaging systems may not be sufficiently accurate to characterize these often asymmetric, complex phenomena. The aim of this study was to validate a three-dimensional (3D) method for determining the severity of aortic regurgitation (AR) in an experimental animal model. METHODS AND RESULTS: In six sheep with surgically induced chronic AR, 20 hemodynamically different states were studied. Instantaneous regurgitant flow rates were obtained by aortic and pulmonary electromagnetic flow meters. Video composite data of color Doppler flow mapping images were transferred into a TomTec computer after computer-controlled 180 degrees rotational acquisition. Direct measurement of the 3D reconstructed FC surface areas as well as measurements of FC areas estimated with 2D methods with hemispherical and hemielliptical assumptions were performed, and values were multiplied by the aliasing velocity to obtain peak regurgitant flow rates. There was better agreement between 3D and electromagnetically derived flow rates than there was between the 2D and the reference values (r=.94, y=1.0x-0.16, difference=0.02 L/min for the 3D method; r=.80, y=1.6x-0.3, difference=1.2 L/min for the 2D hemispherical method; r=.75, y=0.90x+0.2, difference=-0.20 L/min for the 2D hemielliptical method). CONCLUSIONS: Without any geometrical assumption, the 3D method provided better delineation of the FC zones and direct measurements of FC surface areas, permitting more accurate quantification of the severity of AR than the 2D methods. PMID- 9396473 TI - Vesnarinone prolongs action potential duration without reverse frequency dependence in rabbit ventricular muscle by blocking the delayed rectifier K+ current. AB - BACKGROUND: Methanesulfonanilide derivatives, selective inhibitors of the rapidly activating component (I(Kr)) of the delayed rectifier potassium current (I(K)), prolong action potential duration (APD) of cardiac muscles with reverse frequency dependence, which limits their clinical use because of proarrhythmia. Vesnarinone, a quinolinone derivative developed as a cardiotonic agent, has complex pharmacological properties, but its clinical efficacy is explained in part by I(K) reduction. Therefore, we investigated the mode of I(K) block by vesnarinone. METHODS AND RESULTS: I(K) of the rabbit ventricular myocyte was activated by voltage-clamp steps applied from a holding potential to various depolarizing levels. The development of I(K) block at depolarization (+10 mV) and its recovery process at hyperpolarization (-75 mV) were compared between vesnarinone and E-4031. The I(K) block by vesnarinone (3 micromol/L) developed and recovered monoexponentially, with time constants of 361 ms (n=5) and 1.87 seconds (n=4), respectively. I(K) block by E-4031 (0.3 micromol/L) developed instantaneously, with no recovery from the block at hyperpolarization. The I(K) block by vesnarinone, estimated by I(K) tail after a train of depolarizing pulses (for 30 seconds at 0.2 to 2 Hz), was increased with increasing frequency (twofold at 2 from 0.2 Hz), but that by E-4031 was unchanged. In rabbit papillary muscles, vesnarinone (10 micromol/L) prolonged APD at stimulation frequencies >0.2 Hz, whereas E-4031 (0.3 micromol/L) prolonged that in a reverse frequency-dependent manner. CONCLUSIONS: Vesnarinone may prolong the repolarization of human cardiac muscle without reverse frequency dependence, because I(Kr) is expressed in humans as well as in the rabbit. Thus, this drug may be a model for an ideal class III drug without the risk of proarrhythmia. PMID- 9396474 TI - Positive chronotropic actions of parathyroid hormone and parathyroid hormone related peptide are associated with increases in the current, I(f), and the slope of the pacemaker potential. AB - BACKGROUND: The classic calciotropic hormone parathyroid hormone (PTH) and its paracrine factor parathyroid hormone-related protein (PTHrP) both increase heart rate. METHODS AND RESULTS: We used standard electrophysiological techniques to study the effects of PTH and PTHrP on isolated rabbit sinus node, isolated canine Purkinje fibers, and disaggregated rabbit sinus node myocytes. Sinus node maximum diastolic potential, activation voltage, and amplitude were unchanged by PTH or PTHrP (P>.05). However, the slope of phase 4 and the automatic rate were increased at PTH and PTHrP > or = 10 nmol/L (P<.05). Comparable results were seen in canine Purkinje fibers. We then used the perforated-patch technique to study the I(f) pacemaker current in sinus node. PTH 12.5 nmol/L and PTHrP 12.5 to 18 nmol/L increased I(f) at -65 mV by 68+/-41% (n=5) and 69+/-50% (n=5), respectively. Actions of both agents were reversible. The increase in I(f) appeared to result from a change in maximal conductance and not a shift in the voltage dependence of activation. CONCLUSIONS: These observations provide, for the first time, direct electrophysiological support for the chronotropic actions of PTH and PTHrP. They suggest that classic hormones and paracrine factors can have multiple functions and that in the case of PTH and PTHrP, a newly recognized action is to alter automaticity directly. PMID- 9396475 TI - Electrical remodeling due to atrial fibrillation in chronically instrumented conscious goats: roles of neurohumoral changes, ischemia, atrial stretch, and high rate of electrical activation. AB - BACKGROUND: Recently, we developed a goat model of chronic atrial fibrillation (AF). Due to AF, the atrial effective refractory period (AERP) shortened and its physiological rate adaptation inversed, whereas the rate and stability of AF increased. The goal of the present study was to evaluate the role of (1) the autonomic nervous system, (2) ischemia, (3) stretch, (4) atrial natriuretic factor (ANF), and (5) rapid atrial pacing in this process of electrical remodeling. METHODS AND RESULTS: Twenty-five goats were chronically instrumented with multiple epicardial atrial electrodes. Infusion of atropine (1.0 mg/kg; n=6) or propranolol (0.6 mg/kg; n=6) did not abolish the AF-induced shortening of AERP or interval (AFI). Blockade of K+(ATP) channels by glibenclamide (10 micromol/kg; n=6) slightly increased the AFI from 95+/-4 to 101+/-5 ms, but AFI remained considerably shorter than during acute AF (145 ms). Glibenclamide had no significant effect on AERP after electrical cardioversion of AF (69+/-14 versus 75+/-15 ms). Volume loading by 0.5 to 1.0 L of Hemaccel (n=12) did not shorten AERP. The median plasma level of ANF increased from 42 to 99 pg/mL after 1 to 4 weeks of AF (n=6), but ANF infusion (0.1 to 3.1 microg/min, n=4) did not shorten AERP. Rapid atrial pacing (24 to 48 hours; n=10) progressively shortened AERP from 134+/-10 to 105+/-6 ms and inversed its physiological rate adaptation. CONCLUSIONS: Electrical remodeling by AF is not mediated by changes in autonomic tone, ischemia, stretch, or ANF. The high rate of electrical activation itself provides the stimulus for the AF-induced changes in AERP. PMID- 9396476 TI - Mechanisms causing sustained ventricular tachycardia with multiple QRS morphologies: results of mapping studies in the infarcted canine heart. AB - BACKGROUND: Sustained reentrant ventricular tachycardias (VTs) with different QRS morphologies have been observed to occur spontaneously and during programmed stimulation in human hearts. We determined mechanisms that can cause tachycardias with multiple morphologies in a canine model of myocardial infarction by mapping reentrant circuits. METHODS AND RESULTS: Reentrant VT with multiple QRS morphologies was induced in 11 canine hearts with 4-day-old infarcts. Comparison of activation maps of the reentrant circuits in the epicardial border zone associated with each morphology indicated two basic mechanisms. Less frequently, VTs of different morphologies in the same heart were caused by reentrant circuits in different regions of the infarct. Most commonly, the reentrant circuits associated with different morphologies were in the same region. Three different factors caused different exit routes from circuits in the same region, leading to the multiple morphologies. (1) The reentrant wave front for each morphology rotated around the same line of block but in different directions. (2) Reentrant circuits associated with each morphology were similar, but there were small changes in the extent of the central line of block. (3) Reentrant circuits with completely different sizes and shapes caused different morphologies. CONCLUSIONS: In this canine model, tachycardias with multiple morphologies most commonly arise from reentrant circuits in the same region of the infarct, suggesting that most often only one area has electrophysiological properties necessary to sustain reentry. PMID- 9396477 TI - Iridium oxide-coated defibrillation electrode: reduced shock polarization and improved defibrillation efficacy. AB - BACKGROUND: Transvenous implantable cardioverter-defibrillator (ICD) leads are designed to deliver electric shocks to the heart for termination of ventricular dysrhythmias. However, the efficiency of different lead materials has not been well studied. This study compares an ICD lead coated with iridium oxide (IROX), a material that reduces shock-induced polarization, with an otherwise identical, uncoated lead. METHODS AND RESULTS: The defibrillation threshold (DFT) was determined in 13 swine with both IROX-coated and uncoated ICD leads paired with an uncoated "can" electrode. The leads were exchanged through a Teflon sheath to reproduce the intracardiac position. The delivered energy DFT of the IROX-coated lead was 15.9+/-5.4 J and was significantly lower than the delivered energy DFT of the uncoated lead (19.1+/-5.1 J; P<.006). The initial lead impedance was equivalent in both leads (IROX, 41.7+/-5.8 omega; uncoated, 41.3+/-4.7 omega; P=NS) at DFT. However, the impedance rose by 7.3+/-2.0 omega during the first phase and by 3.7+/-2 omega during the second phase with the uncoated lead, whereas the corresponding impedance change was 1.0+/-0.3 omega during phase 1 and 1.6+/-0.5 omega during phase 2 (P<.01 each phase) when the IROX-coated lead was used. CONCLUSIONS: This study shows that an IROX coating of this lead system significantly lowers the DFT energy in the swine model. The blunting of the impedance rise by the IROX coating that is seen is consistent with a reduction in electrode polarization. PMID- 9396479 TI - Myocardial contrast echocardiography: 15 years of research and development. PMID- 9396478 TI - Significance of ventricular myocytes and nonmyocytes interaction during cardiocyte hypertrophy: evidence for endothelin-1 as a paracrine hypertrophic factor from cardiac nonmyocytes. AB - BACKGROUND: In cardiac hypertrophy, both excessive enlargement of cardiac myocytes and progressive interstitial fibrosis are well known to occur simultaneously. In the present study, to investigate the interaction between ventricular myocytes (MCs) and cardiac nonmyocytes (NMCs), mostly fibroblasts, during cardiocytes hypertrophy, we examined the change in cell size and gene expression of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) in cultured MCs as markers for hypertrophy in the neonatal rat ventricular cardiac cell culture system. METHODS AND RESULTS: The size of cultured MCs significantly increased in the MC-NMC coculture. Concomitantly, secretions of ANP and BNP into culture media were significantly increased in the MC-NMC coculture compared with in the MC culture (with the possible contamination of NMC <1% of MC). Moreover, in the MC culture, enlargement of MC and an increase in ANP and BNP secretions were induced by treatment with conditioned media of the NMC culture. A considerable amount of endothelin (ET)-1 production was detected in the NMC-conditioned media. BQ-123, an ET-A receptor antagonist, and bosentan, a nonselective ET receptor antagonist, significantly blocked the hypertrophic response of MCs induced by treatment with NMC-conditioned media. Angiotensin II (Ang II) (10(-10) to 10(-6) mol/L) and transforming growth factor-beta1 (TGF beta1) (10(-13) to 10(-9) mol/L), both of which are known to be cardiac hypertrophic factors, did not induce hypertrophy in MC culture, but both Ang II and TGF-beta1 increased the size of MCs and augmented ANP and BNP productions in the MC-NMC coculture. This hypertrophic activity of Ang II and TGF-beta1 was associated with the potentiation of ET-1 production in the MC-NMC coculture, and the effect of Ang II or TGF-beta1 on the secretions of ANP and BNP in the coculture was significantly suppressed by pretreatment with BQ-123. CONCLUSIONS: These results demonstrate that NMCs regulate MC hypertrophy at least partially via ET-1 secretion and that the interaction between MCs and NMCs plays a critical role during the process of Ang II- or TGF-beta1-induced cardiocyte hypertrophy. PMID- 9396480 TI - Role of the diadic cleft in myocardial contractile control. PMID- 9396481 TI - New look to an old symptom: angina pectoris. AB - At the turn of this century, it was proposed that ischemic cardiac pain might be related to distension of the ventricular wall ("mechanical hypothesis"). Three decades later, it was hypothesized that ischemic pain might be elicited by the intramyocardial release of pain-producing substances induced by ischemia ("chemical hypothesis"). Studies carried out in the past 10 years have given strong support to the chemical hypothesis, because they have consistently shown that adenosine is a mediator of ischemic cardiac pain. Adenosine-induced ischemic cardiac pain is mediated primarily by stimulation of A1 receptors located in cardiac nerve endings and is potentiated by substance P. Conversely, the magnitude and rate of left ventricular dilation during ischemia do not predict the severity of angina. It is worth noting, however, that stretching of epicardial coronary arteries appears to potentiate the severity of angina caused by myocardial ischemia. The nervous activity generated by myocardial ischemia is modulated in intrinsic cardiac, mediastinal, and thoracic ganglia. Then it is further modulated in the central nervous system and projects bilaterally to the cortex, as demonstrated in humans by positron emission tomography, where it is decoded as a painful sensation. The causes responsible for the lack of angina during myocardial ischemia are probably different in patients who present both pain-free and painful myocardial ischemia, in patients with predominantly painless ischemia, and in diabetic patients. PMID- 9396483 TI - Images in cardiovascular medicine. False aortic aneurysm due to rupture of an aortocoronary saphenous vein bypass graft. PMID- 9396482 TI - Premature coronary artery disease associated with a disruptive mutation in the estrogen receptor gene in a man. AB - BACKGROUND: While estrogens protect against coronary artery disease in women, it is unclear whether they influence cardiovascular function in men. The present report describes coronary vascular abnormalities and the lipoprotein profile of a male patient with estrogen insensitivity caused by a disruptive mutation in the estrogen-receptor gene. METHODS AND RESULTS: Stress thallium scintigraphy, echocardiography, and electron-beam computed tomography (CT) scanning of the coronary arteries and detailed lipoprotein analysis were performed. Electron-beam CT scanning of the coronary arteries showed calcium in the left anterior descending artery. Lipoprotein analysis showed relatively low levels of total (130 mg/dL), LDL (97 mg/dL), and HDL (34 mg/dL) cholesterol; apolipoprotein A-I (91.7 mg/dL); and lipoprotein(a) (4.1 nmol/L), but normal levels of triglycerides (97 mg/dL) and pre-beta-1-HDL cholesterol (61 microg/mL). CONCLUSIONS: The absence of functional estrogen receptors may be a novel risk factor for coronary artery disease in men. PMID- 9396484 TI - Images in cardiovascular medicine. Left anterior descending coronary artery-to right ventricle fistula. PMID- 9396485 TI - Does the early hazard phenomenon exist? Do the GUSTO findings prove or disprove? PMID- 9396486 TI - Anger and myocardial infarction. PMID- 9396487 TI - DD genotype of the ACE gene as an atherosclerosis-independent risk factor for myocardial infarction. PMID- 9396488 TI - Aortic regurgitation in women. PMID- 9396489 TI - Minimally invasive heart surgery. PMID- 9396490 TI - Physical activity and plasma viscosity. PMID- 9396491 TI - Coronary heart disease in India. PMID- 9396492 TI - Preserved vasodilatory response to nitrates in saphenous vein bypass grafts. PMID- 9396493 TI - Effects of dobutamine stimulation. PMID- 9396494 TI - Does estrogen replacement prevent coronary heart disease? PMID- 9396495 TI - Pathogenesis of calcification of native and bioprosthetic valves is different. PMID- 9396496 TI - Antimineralization effect of ethanol and experimental model of accelerated calcification study in heart valve bioprostheses. PMID- 9396497 TI - In vitro effects of the platelet glycoprotein IIb/IIIa receptor antagonists c7E3 Fab on the activated clotting time. PMID- 9396498 TI - Nonsustained ventricular tachycardia in severe heart failure. PMID- 9396499 TI - Evidence for the existence of a functional cardiac renin-angiotensin system in humans. PMID- 9396500 TI - Plasminogen activator inhibitor-1 gene and myocardial infarction. PMID- 9396501 TI - Effects of hypoglycemic agents on patients with diabetes. PMID- 9396502 TI - Licensed to prescribe, but....'prior approval required'. PMID- 9396503 TI - Concerning the units for the QT interval corrected by Bazett's formula. PMID- 9396504 TI - Doppler features of constrictive pericarditis. PMID- 9396505 TI - Worry and coronary heart disease. PMID- 9396506 TI - Cholesterol-lowering therapy. PMID- 9396507 TI - Collagen metabolism and restenosis. PMID- 9396508 TI - Common mutation in the methylenetetrahydrofolate reductase gene offers no support for mild hyperhomocysteinemia being a causal risk factor for cardiovascular disease. PMID- 9396509 TI - Oral anticoagulants in patients with mechanical cardiac valve replacement: the role of aspirin. PMID- 9396510 TI - Acoustic assessment of heart valves. PMID- 9396511 TI - Acute profound thrombocytopenia after c7E3 Fab therapy. PMID- 9396512 TI - Ischemia and glucose metabolism. PMID- 9396513 TI - Hypocholesterolemia is associated with low levels of hemostatic risk factors. PMID- 9396514 TI - Electrical remodeling. PMID- 9396515 TI - Endothelin receptor antagonists. PMID- 9396516 TI - Trials in myocardial infarction. PMID- 9396517 TI - Partial ventriculectomy: a promising treatment for patients with end-stage congestive heart failure. PMID- 9396518 TI - The first Stephen L. Gans Memorial Lecture. An international strong personal friendship in pediatric surgery. PMID- 9396519 TI - Total colonic aganglionosis with or without small bowel involvement: a changing profile. AB - BACKGROUND/PURPOSE: To identify recent trends in the diagnosis and treatment of total colonic aganglionosis with or without small bowel involvement (TCSA), the authors analyzed the findings in 107 patients who had TCSA seen between 1988 and 1992 at 147 medical institutions throughout Japan and compared the results with those of a previous survey conducted between 1978 and 1982. RESULTS: The estimated incidence of total colonic aganglionosis was 1 in 58,084 live births, the male to female ratio was 1.5:1, and the incidence of associated anomalies was 15%. These findings were all very similar to those of the previous survey. Ten years ago, 83.6% of all patients underwent Martin's procedure. In the recent survey, this rate had fallen to 52.1%, and a right colon patch method has also been developed as a new procedure. A marked decrease in the overall mortality rate from 40.9% to 21.5% was observed. However, a high mortality rate persists in those cases with small bowel involvement (33.3%); most such cases are complicated with enterocolitis. CONCLUSIONS: Although the general features of TCSA were similar to those in the previous survey, a substantial improvement in the treatment results for TCSA has occurred. However, further efforts are still required both to prevent and more effectively to treat enterocolitis, especially in cases involving any aganglionosis extending into small bowel. PMID- 9396520 TI - Long-term follow-up of patients treated with ileoendorectal pull-through and right colon onlay patch for total colonic aganglionosis. AB - BACKGROUND/PURPOSE: This study was performed to assess the long-term follow-up of five patients who underwent one-stage ileoendorectal pull-through with right colon onlay patch for total colonic aganglionosis (TCA) at Kaiser Permanente Medical Center. METHODS: A retrospective review of inpatient and outpatient charts and telephone follow-up of all patients were conducted to obtain current data regarding growth, development, bowel function, and postoperative and late complications. RESULTS: Follow-up has ranged from 2 to 11 years. All patients are at or above the 50th percentile for weight by age and are continent with 1 to 5 daily bowel movements. Only two patients required reoperation. A perirectal abscess developed in one patient 2 months postoperatively. In the second patient a functional obstruction was relieved by sphincterotomy. CONCLUSIONS: Ileoendorectal pull-through with right colon onlay patch is associated with few early and late postoperative complications; it appears to be superior to other procedures in the early postoperative period because of the more rapid return to acceptable stooling patterns. This method of reconstruction provides an excellent opportunity for normal growth, development, and long-term bowel function. PMID- 9396521 TI - Cytomegalovirus enteritis in a premature infant. AB - BACKGROUND/PURPOSE: Up to 2.5% of newborn infants are cytomegalovirus (CMV) positive at birth. Five percent will be symptomatic at birth, including cytomegalic inclusion disease. Symptoms such as hearing loss and mental retardation will ultimately develop in 15%. METHODS: The authors describe a case of CMV enteritis in a 2.2-kg newborn that presented as necrotizing enterocolitis (NEC) and subsequently developed a colonic stricture. RESULTS: There are four reports of neonatal CMV enteritis in the nonEnglish-language literature. Cytomegalovirus enteritis has become prevalent among the immunosuppressed pediatric and adult patient population. CONCLUSIONS: We propose the addition of CMV to the list of pathogens responsible for NEC. A review of neonatal CMV infection is provided. PMID- 9396522 TI - Factors influencing the outcome of liver transplantation for biliary atresia. AB - BACKGROUND/PURPOSE: This study examined the factors present before liver transplantation (LTx) influencing the outcome in 14 patients who had biliary atresia (BA) who underwent LTx. RESULTS: Nine patients survived (Group A), whereas five died primarily of infection (Group B). Rate of the attempted multiple hepatic portoenterostomy (HPE) and existence of intestinal stoma was significantly higher in Group B than in Group A. Pre-LTx parameters showed significant difference between the two groups as follows: total bilirubin, 15.9 +/- 7.9 versus 29.1 +/- 14.5 mg/dL (P = .0446); gamma-glutamyl transpeptidase, 170.0 +/- 97.6 versus 65.2 +/- 38.8 IU/L (P = .0425); the body weight deviation score, 0.17 +/- 0.88 SD versus -1.46 +/- 0.30 SD (P = .0029); total cholesterol, 129.4 +/- 33.5 versus 52.2 +/- 20.4 mg/dL (P = .0008) in Group A versus Group B. Total cholesterol level and body weight for age remained within normal range until the advanced stage and rapidly decreased according to deterioration of the general condition before LTx. CONCLUSIONS: From these results, avoidance of multiple HPE and closure of stoma before LTx may be preferable. LTx should be performed before failure to thrive or hypocholesterolemia develops. PMID- 9396523 TI - A new hepatic portoenterostomy with division of the ligamentum venosum for treatment of biliary atresia: a preliminary report. AB - BACKGROUND/PURPOSE: Kasai's operation consists of transection of the fibrous portal cord including the bile duct remnant. However, it is difficult to transect the fibrous cord at the level of the posterior surface of the portal vein, because the portal vein is fixed at the porta hepatis. METHODS: The authors described a new hepatic portoenterostomy to transect the fibrous cord under an appropriate visual field by division of the ligamentum venosum (Arantius' canal). Between February and December 1996, six patients who had biliary atresia underwent this procedure. RESULTS: Jaundice resolved completely (TB < or = 1.0 mg/dL) in all six patients within 40 days. Postoperative cholangitis did not occur and good bile drainage was obtained. CONCLUSIONS: Using this procedure, the portal vein becomes fully mobile after dividing the ligamentum venosum, and the porta hepatis can be widely exposed. The fibrous cord of the porta hepatis can be easily dissected off posteriorly and laterally. PMID- 9396524 TI - A new diagnostic approach to biliary atresia with emphasis on the ultrasonographic triangular cord sign: comparison of ultrasonography, hepatobiliary scintigraphy, and liver needle biopsy in the evaluation of infantile cholestasis. AB - BACKGROUND/PURPOSE: The authors evaluated prospectively the utility of ultrasonography, Tc-99m-DISIDA hepatobiliary scintigraphy, and liver needle biopsy in differentiating biliary atresia (BA) from intrahepatic cholestasis in 73 consecutive infants who had cholestasis. METHODS: Sixty three ultrasonographic examinations of 61 infants with 7.0-MHz transducer were carried out, focusing on the fibrous tissue at the porta hepatis. The authors defined the triangular cord (TC) as visualization of a triangular or tubular shaped echogenic density just cranial to the portal vein bifurcation on a transverse or longitudinal scan. RESULTS: Although 17 of 20 ultrasonographic examinations from infants who had BA denoted TC, 43 ultrasonographic examinations from infants with either neonatal hepatitis (NH) or other causes of cholestasis denoted no TC, showing a diagnostic accuracy of 95% with 85% sensitivity and 100% specificity. Investigation with Tc 99m-DISIDA hepatobiliary scintigraphy showed that 24 of 25 infants who had BA had no gut excretion, and 16 of 46 infants who had either NH or other causes of cholestasis had gut excretion, showing a diagnostic accuracy of 56% with 96% sensitivity and 35% specificity. Therefore, gut excretion of tracer excluded BA, but no gut excretion of tracer needed further investigations as liver needle biopsy. Forty-four liver needle biopsies were carried out in 19 infants who had BA and 24 infants who had either NH or other causes of cholestasis. Although 18 of 20 biopsy findings in infants who had BA were correctly interpreted as having BA, 23 of 24 biopsy results in infants who had either NH or other causes of cholestasis were correctly diagnosed, showing a diagnostic accuracy of 93% with 90% sensitivity and 96% specificity. CONCLUSIONS: Since the introduction of ultrasonographic TC sign in the diagnosis of BA by our institution, we have found that it seemed to be a simple, time-saving, highly reliable, and non-invasive tool in the diagnosis of BA from other causes of cholestasis. The authors propose a new diagnostic strategy in the evaluation of infantile cholestasis with emphasis on ultrasonographic TC sign as first priority of investigations. When the TC is visualized, prompt exploratory laparotomy is mandatory without further investigations. When the TC is not visualized, hepatobiliary scintigraphy is the next step. Excretion of tracer into the small bowel actually rules out BA. Liver needle biopsy is reserved only for the infants with no excretion of tracer. The authors believe that a correct decision regarding the need for surgery can be made in almost all cases with infantile cholestasis by this multidisciplinary approach. PMID- 9396525 TI - Initial experience with non-breath-hold magnetic resonance cholangiopancreatography: a new noninvasive technique for the diagnosis of choledochal cyst in children. AB - BACKGROUND/PURPOSE: Magnetic resonance cholangiopancreatography (MRCP) is an emerging tool for the noninvasive evaluation of the pancreaticobiliary tree. METHODS: Non-breath-hold MRCP was used in three children to evaluate choledochal cyst; a first for this new modality of diagnostic imaging. In all cases, the intrahepatic and extrahepatic bile ducts, and the pancreatic duct were clearly visualized. RESULTS: Two cases were found to have a fusiform choledochal cyst, and non-breath-hold MRCP demonstrated pancreaticobiliary malunion and a long common channel. In the remaining case, the size and location of the huge cyst prevented visualization of any pancreaticobiliary malunion. Endoscopic retrograde cholangiopancreatography (ERCP) in this patient failed to provide any additional information. All patients underwent cyst excision with hepaticoenterostomy, and made an uneventful recovery. CONCLUSIONS: Our initial experience suggests that non-breath-hold MRCP is a reliable method for the diagnosis of choledochal cyst in children and could replace ERCP. PMID- 9396526 TI - Early and late results of excision of choledochal cysts. AB - BACKGROUND/PURPOSE: Reports on the late results of choledochal cyst excision with hepaticojejunostomy in children are relatively few. METHODS: Of the 84 patients who had choledochal cyst who came under our care, 79 have had definitive surgery, three are awaiting surgery, one is being observed with Caroli's disease, and the parents of one child have refused surgery. Thirty-eight patients treated decades ago had internal drainage procedures. Since 1972, 41 patients have had cyst excision with hepaticojejunostomy using a 40-cm Roux loop without an antireflux procedure. Early complications in those who underwent cyst excision with hepaticojejunostomy included anastomotic leak in three patients who required reoperation, cholangitis in two, and fluid collection in the gall-bladder bed that required no intervention in one. RESULTS: During a follow-up period ranging from 4 months to 17 years (mean, 8.5 years), anastomotic stricture, cholangitis, and intrahepatic stone formation developed in two children after being well for 8 years and over 11 years. These children required additional surgical procedures to overcome their problems. Asymptomatic intrahepatic stones 2 years after cyst excision with hepaticojejunostomy developed in a third child. There was no mortality in the entire group that underwent cyst excision and they are all enjoying a good quality of life. CONCLUSIONS: Careful, long-term follow-up is important in children who have choledochal cyst excision with hepaticojejunostomy. PMID- 9396527 TI - Two cases of torsion of the gallbladder diagnosed preoperatively. AB - BACKGROUND: The authors experienced two cases of torsion of the gallbladder, both of which were correctly diagnosed preoperatively. METHODS: Two boys, aged 4 and 5 years, were transferred to Yokohama City Seibu Hospital because of their acute abdominal disorders. Diagnostic imaging demonstrated acute inflammatory changes in the gallbladder with an abnormal arrangement of the gallbladder and common bile duct in both cases. Laparotomy findings showed torsion of the gallbladder by 180 degrees and 360 degrees, respectively, at the cystic ducts, resulting in gangrenous change. RESULTS: Both children recovered uneventfully after cholecystectomy. The diagnostic imaging criteria are (1) collection of fluid between the gallbladder and the gallbladder fossa of the liver, (2) a horizontal rather than vertical arrangement of the long axis of the gallbladder, (3) the presence of a well-enhanced cystic duct located on the right side of the gallbladder, and (4) signs of inflammation including marked edema with thickening of the wall. CONCLUSIONS: The authors report the clinical characteristics of this uncommon condition, and discuss the significance of accurate preoperative diagnosis of these acute surgical disorders. PMID- 9396528 TI - Esophageal atresia in Osaka: a review of 39 years' experience. AB - BACKGROUND: One hundred fifty-nine patients who had esophageal atresia with or without tracheoesophageal fistula have been treated at Osaka University Medical School and its affiliated hospitals since the initial (Japanese) experience of Dr T. Ueda in 1957. METHODS: These cases were divided chronologically into three groups. With earlier recognition of surgical neonates and the development of perinatal care, the long-term survival of these patients has steadily improved over 39 years from 28% in the first period (1957 to 1967) to 80% in the third period (1980 to 1995). Of 141 patients treated in the second and third periods (1968 to 1995), 92 (65.2%) had associated anomalies. Cardiovascular and gastrointestinal malformations were the most frequently seen major anomalies. VATER or VACTER association was seen in 12.8% (18 of 141) of these patients. Survival of these cases according to Waterston risk factors was 100% for group A, 100% for group B, and 50% for group C, whereas the new classification proposed by Spitz showed survival of 92% for group 1, 50% for group 2, and 0% for group 3, showing better differentiation among the three groups. RESULTS: There was a long gap between the proximal and distal esophageal ends in seven patients (type A), in all of whom primary anastomosis was possible after 28 to 128 days of elongation by bouginage. Although the survival of esophageal atresia patients dramatically improved in recent years, there is still a high incidence of early and long-term postoperative complications, ie, anastomotic leakage (26.5%), recurrent fistula (7.2%), anastomotic stricture (49.1%), postoperative pneumonia or atelectasis (57.0%), tracheomalacia (25.8%), and gastroesophageal reflux (52.0%). CONCLUSIONS: Recently, there have been changing patterns in the occurrence of complications, which are mainly attributed to technical improvement, better perinatal care and early recognition of pathophysiologic conditions such as tracheomalacia and gastroesophageal reflux. PMID- 9396529 TI - Tracheomalacia with esophageal atresia and tracheoesophageal fistula in fetal rats. AB - BACKGROUND: Many patients who have esophageal atresia and tracheoesophageal fistula (EA-TEF) have associated tracheomalacia, which is thought to be one of the reasons for respiratory complications after surgical correction of the abnormality. METHODS: In this study, tracheas from Adriamycin-induced EA-TEF fetal rats were examined histologically and relevant cross-sectional parameters of the tracheas were measured. RESULTS: The tracheal lumen in tracheomalacia was small and irregular, losing its normal "D" shape. In most rats, the cartilaginous ring was broken into two to four segments, making the trachea lose its rigid support. The submucosa was thickened with prominent bulging of its membranous part into the tracheal lumen. The ratio of the inner luminal cross-sectional area to the outer tracheal cross-sectional area in EA-TEF rats was 15.7%, compared with a control ratio of 47.2%. In EA-TEF rats, the length of the cartilaginous ring was significantly shortened (P < .001), but not the length of membranous trachea, thus resulting in a cartilaginous/membranous (C/M) ratio of 1.55:1, markedly lower than that of normal rats (4.34:1, P < .001). The reduction of anterior-posterior diameter of the tracheal lumen was more marked than that of the transverse diameter. CONCLUSIONS: These observations suggest that the trachea in EA-TEF rats has a smaller lumen and is more flaccid than normal, making it prone to airway obstruction. The fact that tracheomalacia developed only in fetuses who had EA-TEF indicates that the factors that result in EA-TEF also cause tracheomalacia. PMID- 9396530 TI - The vagus and recurrent laryngeal nerves in the rodent experimental model of esophageal atresia. AB - BACKGROUND: After surgical correction of their esophageal atresia and tracheoesophageal fistula (EA-TEF), many patients exhibit evidence of esophageal dysmotility. Controversy exists as to whether the esophageal motility disorders result from denervation caused by surgery or from an inherent abnormal innervation of the esophagus. METHODS: The present study used an Adriamycin induced EA-TEF fetal rat model to trace the course and branching of both the vagus and recurrent laryngeal nerves. Abnormalities observed in EA-TEF rat fetuses include: (1) fewer branches from both recurrent laryngeal nerves; (2) deviation of the left vagus from its normal course below the aorta, passing behind the fistula to approach and join with the right vagus to form a single nerve trunk on the right side of the esophagus; (3) relatively few branches from the single vagal nerve trunk (composed of fibers of the left and the right vagus) on the surface of the lower esophagus. CONCLUSIONS: Fetuses affected by EA-TEF have inherent abnormalities in the course and branching pattern of the vagus nerves as they descend through the thorax, culminating in a deficient extrinsic nerve fiber plexus in the lower esophagus. These observations may account for the esophageal motility disorders seen in patients who have EA-TEF even before surgical intervention. PMID- 9396531 TI - Aortic arch anomalies associated with long gap esophageal atresia and tracheoesophageal fistula. AB - PURPOSE: The purpose of this study was to determine whether aortic arch anomalies are associated with long gap esophageal atresia and tracheoesophageal fistula (EA TEF). METHODS: The authors performed a retrospective review of all infants who had EA-TEF from 1980 to 1996 at two pediatric surgery centers. Two hundred three infants who had EA-TEF were identified. RESULTS: Twelve infants were noted to have both long gap EA-TEF defined as a gap length greater than 3 cm and aortic arch anomalies. Of these 12, 7 had aberrant right subclavian arteries originating from the descending aorta. Four of the seven infants who had aberrant right subclavian artery (SCA) had gap lengths greater than 4 cm. All four had their fistulae divided initially through a right thoracotomy with primary repair performed at a later date. The remaining five infants who had long gap EA-TEF had right-sided aortic arch with aberrant left subclavian arteries. All five initially underwent exploration through the right chest. On discovery of the long gap EA and concurrent vascular anomaly, the thoracotomies were closed, and the infants underwent definitive repair of both their EA-TEF and their vascular anomaly through a left thoracotomy. CONCLUSIONS: The authors find that aortic arch anomalies are associated with long gap EA-TEF. Patients who have these two anomalies tend to have a long gap. Preoperative diagnosis of these anomalies may alter the timing and technique of surgical intervention. The embryogenesis of these vascular lesions may account for this more severe form of esophageal atresia. PMID- 9396532 TI - Congenital true diverticula of the esophagus: a case report. AB - Pseudodiverticulosis secondary to gastroesophageal reflux is a common disease in adults, but true esophageal diverticula are rare in infants and children. A 5 year-old boy was well until the age of 1 1/2 years when he started vomiting. An upper gastrointestinal series showed two diverticula bulging from the posterior right side of the middle esophagus associated with slight hiatal hernia and short esophagus. Diverticulectomy, the Collis-Nissen antireflux procedure, and pyloroplasty were performed simultaneously through a left thoracoabdominal incision. Histological examination of the diverticula showed that the wall of each diverticulum consisted of a full-thickness of esophageal wall. Because there was no tracheal remnant in the diverticula, this lesion is more likely to be a true diverticulum than a duplication. PMID- 9396533 TI - Selecting the surgical procedure for simple and complicated esophageal achalasia in children. AB - BACKGROUND/PURPOSE: Surgical experience in children who have achalasia is limited. Surgical treatment requires esophagocardiomyotomy and an antireflux procedure. However, when these operations fail, other procedures are needed. To summarize the experience treating children who have this condition, the authors reviewed retrospectively all case histories of patients treated from 1971 to 1996 at their hospital. METHODS: Three boys and a girl, ranging in age from 18 months to 11 years, were treated. All had multiple previous dilatations. Two then underwent operation using an abdominal approach for a Heller procedure and a posterior fundoplasty (Guarner operation). Two children were previously treated in another hospital. One underwent a Heller operation complicated by perforation of the anterior mucosa. The other had undergone three previous abdominal approaches for esophagocardiomyotomy and a Nissen fundoplication. Symptoms persisted and imaging and endoscopy showed stenosis in both patients. In the first patient an esophagocardioplasty with transverse closure (Wendel procedure) and a posterior fundoplasty was performed. In the second child, the three previous abdominal surgical approaches mandated a transthoracic approach with transdiaphragmatic latero-lateral esophagogastric anastomosis (Heyrowsky operation) and a modified Guarner operation using the remaining fundus and gastric body. RESULTS: There were no intraoperative or postoperative complications. Follow-up time ranged from 3 months to 17 years. All patients experienced dramatic relief of symptoms and satisfactory weight gain. No recurrence of symptomatology has occurred. CONCLUSIONS: Esophagocardiotomy associated with an antireflux procedure may be the first option in the surgical treatment of children who have achalasia. However, if this fails, esophagocardioplasty and the latero-lateral esophagogastric anastomosis associated with antireflux procedure may be successful alternatives. PMID- 9396534 TI - Growth factor enhancement of intestinal function: dramatic response but lack of synergism. AB - BACKGROUND/PURPOSE: The authors have shown that gastrin and epidermal growth factor (EGF) exert a trophic effect on intestinal epithelial cells. Because these peptides may have different mechanisms by which they stimulate these cells, this study was designed to determine the effect of gastrin and EGF on the intestinal epithelial cell and to evaluate their potential synergistic effect. METHODS: Twenty young adult male Sprague-Dawley rats underwent placement of jugular venous catheters that were connected to subcutaneously placed osmotic minipumps. The rats were divided into four groups based on the content of the osmotic pump: Group 1, saline (control, n = 5); Group 2, EGF at 150 microg/kg/d (n = 5); Group 3, gastrin at 13.5 nmol/kg/d (n = 5); and Group 4, EGF at 150 microg/kg/d plus gastrin at 13.5 nmol/kg/d (n = 5). After a 14-day intravenous infusion, [C14] galactose and [C14] glycine absorption (pmol/cm2 intestine), mucosal DNA content (microg/mg mucosa), and protein content (microg/mg mucosa) were measured in the small intestine of each rat. RESULTS: The galactose absorption, glycine absorption, DNA content, and protein content were significantly increased by EGF (69%, 28%, 64%, and 55%, respectively) and gastrin (72%, 60%, 93%, and 48%, respectively) when compared with control. Combining EGF and gastrin also significantly increased these parameters (61%, 44%, 96%, and 70%, respectively) when compared with control. However, these data demonstrate no further enhancement than the effect of each peptide alone. CONCLUSION: EGF and gastrin individually may be useful for patients who have inadequate intestinal function, but when combined did not exert a synergistic benefit. PMID- 9396535 TI - Should laparoscopic appendectomy be avoided for complicated appendicitis in children? AB - BACKGROUND/PURPOSE: Laparoscopic appendectomy is becoming the preferred technique for treating acute appendicitis. However, recent literature on adults suggests that laparoscopic appendectomy may increase the risk for postoperative infectious complications in complicated (gangrenous or perforated) cases. This study was undertaken to compare the results of open versus laparoscopic appendectomy for complicated appendicitis in children. METHODS: A retrospective review from two institutions was performed for all children treated operatively for complicated appendicitis from January 1994 through November 1996. RESULTS: Fifty-six cases were identified. Twenty-seven children underwent laparoscopic appendectomy, whereas 22 underwent open appendectomy. Seven children underwent conversion from laparoscopic to open surgery. Operating times and length of hospital stay did not differ significantly between the laparoscopic and open groups. Postoperative complications developed in 24 children (42.8%). Complications were more frequent after laparoscopic appendectomy compared with open appendectomy (56% v 18%, P = .002). A postoperative intraabdominal abscess (IAA) developed in 14 children (25%). An IAA occurred in two children after open appendectomy compared with 11 children after laparoscopic appendectomy (9% v 41%, P = .01). CONCLUSION: The findings suggest that laparoscopic appendectomy should be avoided in children who have complicated appendicitis because of the increased risk for postoperative intraabdominal abscesses. The authors propose a prospective, randomized trial to verify this finding. PMID- 9396536 TI - Cost-effectiveness in diagnosing infantile hypertrophic pyloric stenosis. AB - PURPOSE: The purpose of this study was to determine which imaging study, upper gastrointestinal series (UGI) or abdominal ultrasonography (US), is more cost effective in diagnosing infantile hypertrophic pyloric stenosis (IHPS) using a decision analysis model. METHODS: Probabilities were calculated from a review of the records of all infants less than 6 months of age referred for UGI or US to rule out IHPS over a 3-year period from January 1992 to December 1995. Cost effectiveness was determined from hospital charges for each imaging study and its possible outcomes. RESULTS: The positive predictive value of UGI was 1.0 and US was 0.98 in the 246 infants evaluated for possible IHPS. In patients who had an initially normal study finding (UGI or US), 25% of patients undergoing US first required a second study for persistent symptoms, whereas only 6% of patients who had a negative initial UGI finding required a second study. CONCLUSIONS: Cost analysis found UGI to be more cost-effective than US because fewer secondary studies were required. UGI provides information regarding other pathological conditions as compared with US. PMID- 9396537 TI - Long-term follow-up of childhood duodenal ulcers. AB - PURPOSE: This study reports the long-term results in children who have duodenal ulcers diagnosed by endoscopy who were treated with H2-receptor antagonist. METHODS: The medical records of 32 children admitted into The Queen Mary Hospital with endoscopically proven duodenal ulcers between 1975 and 1988 were reviewed to evaluate the long-term outcome of childhood duodenal ulcers after initial treatment with H2-receptor antagonist (H2RA). Follow-up details were updated and patients who had been lost to follow-up were recalled. The age of the 22 boys and 10 girls at the time of diagnosis of the ulcers ranged from 3 to 16 years (mean, 11.8 yrs). The duration of follow-up ranged from 8.5 to 21 years (mean, 11.6 yrs). RESULTS: Their primary presentations included epigastric pain (n = 9, 28.0%); nonsteroidal antiinflammatory drug (NSAID)-induced gastrointestinal bleeding (GIB, n = 6, 18.7%); unprovoked GIB (n = 12, 37.5%); perforation (n = 4, 12.5%); and pyloric obstruction (n = 1, 3.0%). All 13 patients who had NSAID induced ulcers (pain and bleeding) responded to H2RA therapy and required no further treatment. All 14 patients who had unprovoked ulcers who presented with pain or bleeding did not respond to H2RA treatment. Ulcer healing was achieved only after eradication of Helicobacter pylori with antibiotics (n = 8) or definitive surgery involving either truncal vagotomy and pyloroplasty (VP, n = 4) or proximal gastric vagotomy (PGV, n = 2). The patient who had gastric outlet obstruction had vagotomy and antrectomy. All four patients who had perforation were initially treated with patch repair, but two had persistent ulceration despite H2RA treatment and required PGV. Complications developed in none of the four patients who had PGV, whereas two of the four patients with VP had problems (diarrhea, n = 1; bezoar obstruction, n = 1). CONCLUSIONS: Unprovoked childhood duodenal ulcer is associated with significant long-term morbidity and requires continued follow-up. The majority of the ulcers are resistant to H2RA treatment alone and ultimately require either eradication of H. pylori or surgery. In the absence of obstruction, PGV may be enough to resolve the ulcer diathesis. PMID- 9396538 TI - Pancreatoblastoma. AB - BACKGROUND: Pancreatoblastoma is a rare pancreatic tumor with distinct acinar and squamoid cell differentiation that generally affects infants and young children. Just over 50 cases have been reported in the literature. METHODS: Five cases of pathologically proven pancreatoblastoma treated at Seoul National University Hospital from 1984 to 1994 were reviewed. There were three girls and two boys who were 2 years to 5 years of age. All cases came to medical attention because of an abdominal mass. RESULTS: Abdominal pain was observed in one case and anorexia, vomiting, and weight loss in one case. There was marked elevation of serum alpha fetoprotein (27,000 ng/mL) in one case of liver metastases. Complete excision was performed in two cases in which the tumors were located in the tail of the pancreas. Partial excision was performed in two patients who had unresectable tumors of the head of the pancreas. One patient had an unresectable tumor at diagnosis and needle aspiration biopsy was carried out under ultrasound guidance. Electron microscopy was performed on pathological specimens of three cases and showed zymogen granules but not neuroendocrine granules. Immunocytochemical studies for alpha-fetoprotein, insulin, glucagon and somatostatin were performed in one patient, and results were all negative. Of two patients who underwent complete excision, one patient presented with liver metastases 4 months after operation and received chemotherapy, but died of tumor 6 months after operation. The other patient had local recurrence 1 year after operation. Reoperation and chemotherapy were performed, and the child is now alive without evidence of disease for 32 months. All three patients who had unresectable tumor died of tumor despite adjuvant radiotherapy and chemotherapy. CONCLUSIONS: The authors emphasize that the diagnosis of pancreatoblastoma in childhood should be suspected with palpation of an abdominal mass, and the chance for cure may be determined by complete excision of the tumor. PMID- 9396539 TI - Is extensive surgery required for treatment of advanced neuroblastoma? AB - BACKGROUND: Prognosis of advanced neuroblastoma is still disappointing although recently slightly improving because of more intensive chemotherapy supported with stem cell transfusion. Before 1985, all patients at University of Tsukuba over the age of 1 year who had stage III and IV neuroblastoma died regardless of extensive resection of the primary tumor. METHODS: Since the treatment protocol of the Study Group of Japan for Advanced Neuroblastoma was introduced in 1985, the authors treated 14 consecutive patients over the age of 1 year who had advanced neuroblastoma with six to eight cycles of the intensive induction chemotherapy regimens followed by resection of the primary and local lymph node metastasis combined with intraoperative irradiation. The resection of the primary tumor and the lymph node metastasis was much less extensive preserving adventitia with perivascular nerves to avoid postoperative vascular occlusion, intestinal dysmotility, and massive lymphorrhea, which interfere with postoperative intensive chemotherapy. If the dissection of lymph nodes from major vessels was difficult, the authors intentionally left the tumor-containing lymph nodes. RESULTS: There were macroscopic residual tumors in 8 of 14 patients. Electron beam irradiation, 10 to 15 Gy, was given to the tumor bed intraoperatively. Overall survival of these 14 patients was 63% (eight patients) at 5 years with six patients surviving without recurrence for more than 5 years. Five patients died of tumors, two of whom died before surgery. Local tumor control failed in only two patients. In one patient, the tumor recurred twice 47 months and 61 months after therapy. After undergoing a second resection at 69 months, this child has survived tumor free for 12 months after the second recurrence. The other patient who had tumor recurrence had an N-myc-amplified tumor that recurred 4 months postoperatively in the hepatoduodenal ligament locally with massive bone metastasis. The 5-year local relapse-free probability for patients with stage III and IV tumors who had an operation was 79% by the Kaplan-Meier method. CONCLUSION: Systematic extensive surgery for advanced or metastatic neuroblastoma is no longer required if supplemented with intensive pre- and postoperative chemotherapy with intraoperative radiotherapy. PMID- 9396540 TI - Histopathologic findings of advanced neuroblastoma after intensive induction chemotherapy. AB - BACKGROUND: Histopathologic findings of advanced neuroblastoma after intensive induction chemotherapy have not been studied well. METHODS: In the present study, all of the surgical specimens from 19 patients who had advanced abdominal neuroblastoma and were pretreated intensively with the protocol of the Study Group of Japan were reviewed. The authors found that dissection of the contralateral lymph nodes is mandatory in advanced neuroblastoma when the goal is the complete dissection of the abdominal disease. Effects of chemotherapy were graded histologically according to the ratio of viable residual neuroblastoma tissue to total areas of the tumor, including neuroblastoma, ganglioneuroblastoma, ganglioneuroma, hemorrhage, necrosis and fibrosis, in five ranks from ( ) to (-). CONCLUSIONS: The newly introduced, highly cytotoxic regimen of the Japanese protocol, designated "A3," appears to be more effective histologically than the conventional regimen, designated "A1" or "new A1." Effects designated ( ) or (++) were prerequisites for survival in stage IV disease, but some stage III patients with the (+) effect survived. PMID- 9396541 TI - Needle localization for thoracoscopic resection of small pulmonary nodules in children. AB - BACKGROUND: Children who have malignant disease and pulmonary nodules frequently need a tissue diagnosis to direct therapy. Computed tomography (CT)-guided needle localization and methylene blue marking allow thoracoscopic resection of nonvisible nodules. METHODS: Malignant disease was diagnosed in three patients aged 2, 2.5, and 11 years. Pulmonary nodules seen on chest CT, representing either metastatic disease or infection developed in each patient. All lesions were 1 to 2 cm deep to the pleural surface, precluding thoracoscopic visualization. A Homer mammographic needle was placed near the lesion using CT guidance under general anesthesia. The pleura overlying the lesion was also marked with methylene blue. Under the same anesthetic, patients went to the operating room where the lesions were thoracoscopically resected. RESULTS: Needle localization and methylene blue staining accurately localized the lesion in all cases. Thoracoscopic resection provided a diagnosis of metastatic disease or infection in all cases. There were no complications. CONCLUSION: CT-guided needle localization of pulmonary lesions deep to the pleural surface, is a safe, accurate method for allowing thoracoscopic resection in these children who would otherwise need open thoracotomy for diagnosis. PMID- 9396542 TI - Is a new biofeedback therapy effective for fecal incontinence in patients who have anorectal malformations? AB - PURPOSE: The authors devised computerized equipment for use in the biofeedback therapy in the management of fecal continence after surgery for anorectal malformations. METHODS: The therapy was used for two to eight sessions in 14 children (11 who had high-type anomalies and three who had intermediate-type anomalies). The ages ranged 5 to 14 years. A control group of 17 children, aged 5 to 11 years, who had encopresis, was also treated with the same biofeedback therapy. RESULTS: Clinical improvement was noted in 5 of the 14 (36%) children who had fecal incontinence, and in 15 of the 17 children (88%) who had encopresis. Both in patients who had fecal incontinence and in those who had encopresis, anal resting pressures were not affected by biofeedback therapy. Furthermore, the anal resting pressure in children who had fecal incontinence was significantly lower than that in children who had encopresis. However, anorectal manometry showed that the biofeedback therapy improved voluntary sphincter function and rectal sensation in both groups. CONCLUSION: Biofeedback therapy appears to be effective in most children who have encopresis whose sphincter function is intact, and in some children who have fecal incontinence after surgery for anorectal malformations. PMID- 9396543 TI - Continent appendicostomy in the bowel management of fecally incontinent children. AB - BACKGROUND: Fecal incontinence is common in children who have anorectal malformations, Hirschsprung's Disease, and spina bifida and can negatively impact their emotional and social development. Enemas have been used as an artificial way to keep children clean and to improve their quality of life. This method is unpleasant for many children, particularly when they reach adolescence. Malone in 1990 described an alternative method in which the appendix is used as a conduit to administer an antegrade enema. METHODS: The authors describe their experience with this new procedure, modified by them, and used in 20 patients. In the original procedure, the base of the appendix is divided, inverted, and reimplanted into the cecum with an antireflux technique. The authors simplify this by plicating the cecum around the appendix to create a one-way valve mechanism but leaving the appendix in its original position. The authors also mobilize the cecum and exteriorize the appendix at the umbilicus to create an inconspicuous stoma. If the native appendix is absent a neoappendix was created from a flap of cecum. RESULTS: Nineteen of 20 patients (95%) are now completely clean 24 hours a day. Stricture of the stoma occurred in two patients and required revision. Leakage at the appendicostomy site occurred in three patients, and two required a tighter plication. CONCLUSIONS: The technique is used to change the route of enema administration, and is only used in patients for whom bowel management with rectal enemas has been proven successful. The appendix must be preserved whenever possible in patients at risk for fecal incontinence. PMID- 9396544 TI - Prospective analysis of lung-to-head ratio predicts survival for patients with prenatally diagnosed congenital diaphragmatic hernia. AB - BACKGROUND: Accurate prenatal prediction of outcome for fetuses who have congenital diaphragmatic hernia (CDH) is very difficult. The authors previously reported a retrospective analysis of risk factors for fetal CDH and proposed a new index of severity: the lung-to-head ratio (LHR). The authors now report a prospective study to test whether this new index predicts neonatal outcome. METHODS: Fifteen patients who had left-sided CDH were sonographically evaluated at the University of California, San Francisco (UCSF) and followed prenatally and postnatally. LHR was measured at 24 to 26 weeks' gestation. Outcome variables included survival and the need for extracorporeal membrane oxygenation (ECMO). RESULTS: Overall survival was 47%. LHR ranged from 0.62 to 1.86. No patient with an LHR of less than 1.0 (n = 3) survived despite ECMO, whereas all patients with an LHR greater than 1.4 survived (n = 4), one requiring ECMO. LHR values between 1.0 to 1.4 were associated with 38% survival (n = 8), 75% requiring ECMO. Overall, survivors had a mean LHR of 1.4 +/- 0.33 and nonsurvivors, 1.05 +/- 0.3 (P < .05). CONCLUSION: The LHR is a useful index to help predict neonatal outcome in patients who have left-sided CDH. PMID- 9396545 TI - Correction of congenital diaphragmatic hernia in utero VII: a prospective trial. AB - BACKGROUND: Congenital diaphragmatic hernia (CDH) remains an unsolved problem. Despite optimal postnatal care, up to 60% of CDH babies die. Experimental evidence and clinical experience have shown that in utero repair of CDH is feasible and can reverse pulmonary hypoplasia, but only in fetuses without liver herniation. For this subgroup, the safety and efficacy of repair before birth has not been compared with standard care after birth. METHODS: Four fetuses in whom CDH without liver herniation was diagnosed underwent open fetal surgery for repair of the CDH. Seven comparison fetuses were treated conventionally. Neonatal mortality was the principle outcome variable. Secondary outcome variables included death of all causes until 2 years of age, number of days of ventilatory support, length of hospital stay, requirement for extracorporeal membrane oxygenation (ECMO), and total hospital charges. RESULTS: There was no difference in survival between the fetal surgery group and the postnatally treated comparison group (75% v 86%). Fetal surgery patients were born more prematurely than the comparison group (32 weeks v 38 weeks' gestation). Length of ventilatory support and requirement for ECMO were equivalent in the fetal surgery group and the postnatally treated comparison group. Length of hospital stay and hospital charges did not differ between the groups. CONCLUSIONS: Open fetal surgery is physiologically sound and technically feasible, but does not improve survival over standard postnatal treatment in the subgroup of CDH fetuses without liver herniation, primarily because overall survival in this subgroup is favorable with or without prenatal intervention. These data suggest that fetuses who have prenatally diagnosed CDH and without evidence of liver herniation should be treated postnatally. PMID- 9396546 TI - Diaphragmatic eventration in infants and children: is conservative treatment justified? AB - PURPOSE: The purpose of this study is to examine the justification of diaphragmatic plication to treat diaphragmatic eventration. A retrospective review of 50 patients who underwent diaphragmatic plication for phrenic nerve injury (PNI) or congenital muscular deficiency (CMD) of the diaphragm was conducted. METHODS: During the last 26 years, 50 patients, aged 4 days to 7 years, were surgically treated for diaphragmatic eventration. Twenty-five patients had iatrogenic PNI and another 25 had CMD. Respiratory distress developed in all patients who had PNI and 10 required mechanical ventilatory support for 13 to 78 days (mean, 41 days) before operation. Respiratory symptoms developed in 17 of 25 patients who had CMD, and four required ventilatory support. In those who were asymptomatic, we justified surgical repair to optimize future lung growth. All patients underwent diaphragmatic plication by a thoracic approach. Reefing mattress sutures on pledgets were used for the plication. RESULTS: In patients who had PNI, ventilatory support could be discontinued within 0 to 6 days (mean, 3 days) after operation, with a dramatic improvement in their respiratory status. Two patients required reoperation because the plication was not tight enough. Seven patients died in this series, but none because of the diaphragmatic plication. CONCLUSION: This study suggests that symptomatic patients who have diaphragmatic eventration should be operated on immediately with an expected dramatic resolution of their respiratory problems. PMID- 9396547 TI - Extracorporeal life support for nonimmune hydrops fetalis. AB - BACKGROUND/PURPOSE: Most babies born with idiopathic nonimmune hydrops fetalis (NIHF) suffer generalized cardiopulmonary collapse and die despite maximal medical therapy. With reported survival rates of less than 10%, many centers consider NIHF an unsalvageable situation and the babies who have this condition, untreatable. In this study, the authors questioned if the aggressive use of extracorporeal life support (ECLS) could salvage this condition and improve the chances of survival for babies born with NIHF. METHODS: The Extracorporeal Life Support Organization's (ELSO) neonatal registry was searched for all available information on babies treated for hydrops fetalis. The ELSO records of all hydropic babies were then reviewed to exclude those babies who had identifiable causes of hydrops. Survival statistics were then calculated for the remaining core group of idiopathic NIHF babies before separating them into two groups based on survival. A detailed analysis comparing the survivors with nonsurvivors was then performed. RESULTS: A total of 28 hydropic babies were identified in the ELSO registry. Four babies were excluded from analysis because of identifiable causes of hydrops (two with congenital diaphragmatic hernia, one with Rh incompatibility, and one with fetal anemia). Of the remaining 24 babies who had NIHF, 54% (13 babies) survived the neonatal period and were discharged from the hospital. Analysis comparing the survivors with the nonsurvivors in our study showed that the groups were similar in their gestational ages, birth weights, Apgar scores and the time to initial intubation. The most distinguishing factor of survival in our study was that the survivors, on average, received ECLS support 3 days sooner than nonsurvivors (mean, 17.5 +/- 1.3 hours of life for survivors v 105 +/- 36.6 hours for nonsurvivors, P < or = .05). CONCLUSION: Idiopathic NIHF should no longer be considered an untreatable condition but a new indication for ECLS that, when begun early, may significantly improve the chances of survival for these babies previously considered "unsalvageable." PMID- 9396548 TI - Ultrasonographic study of the sternocleidomastoid muscle in the management of congenital muscular torticollis. AB - BACKGROUND: Congenital muscular torticollis (CMT) in infancy is caused by the fibrotic change of the sternocleidomastoid muscle (SCMM). The etiology and management strategies remain controversial. METHODS: One hundred ninety-seven infants and children aged 1 month to 16 years who had CMT were examined by real time ultrasonography of the SCMM between June 1995 and September 1996 in a prospective and longitudinal study. A total of 362 examinations were performed. There were 122 boys and 75 girls. RESULTS: The right side was involved in 117 patients (59.3%), the left side in 79 patients (40.1%), and both sides in one patient. The sonographic findings were homogeneous or heterogeneous (patchy) hyperechoic lesion within the SCMM, and all were diagnostic. The ultrasonographic appearance of the SCMM in this study has a close resemblance to the clinical course of CMT. The extent of fibrosis as represented by the cross section of lesion to muscle ratio (L/M ratio) decreased from 83.6% at 2 months to 59.9% at 9 months of age and further decreased to 40% beyond 1 year of age. This consistent decrease in fibrosis was caused by the increased normal muscle volume at the periphery and by the regenerated muscle fibers within the lesion. In this series of 197 patients, 32 (16.2%) eventually underwent surgery to release the SCMM because of persistent head tilt, chin deviation and limited range of neck motion beyond 1 year of age. The L/M ratio of the operative group was 62.7 +/- 16.0% compared with an L/M ratio of 54.5 +/- 14.2% (P = .035) for the nonoperative group at 1 year of age. The extent of fibrotic change in the cross section of the muscle was a significant factor in determining prognosis. In the longitudinal section, the fibrotic change was limited to only the lower third of the SCMM in 27 patients, and all of them recovered without operation. In 95 patients, the fibrotic lesion was limited to the middle and lower third or middle third only, and only six (6.3%) underwent operation. However, in 75 cases the entire length of muscle was involved, and 26 (34.7%) required surgical release of the contracted muscle. Whole-length muscle involvement was also important for predicting recovery without operative intervention. CONCLUSIONS: Ultrasonographic study of the SCMM is not only a valuable diagnostic tool but can also serve as a useful guideline for the treatment of infants who have congenital muscular torticollis. PMID- 9396549 TI - A comparison of the effect of growth factors on intestinal function and structure in short bowel syndrome. AB - BACKGROUND/PURPOSE: Epidermal growth factor (EGF) and Insulin like growth factor 1 (IGF-1) increase substrate absorption beyond the normal adaptive response after massive small bowel resection in the rat. However, the mechanism for this response is unknown. This study was designed to evaluate the ultrastructural features of the rat small intestine epithelium after exposure to EGF and IGF-1 and correlate any changes with a possible hypothesis regarding the mechanism for the increased absorption. METHODS: Male Sprague-Dawley rats underwent an 80% small bowel resection and jejunostomy tube placement. Seven days later an osmotic pump placed subcutaneously and containing the test substance was connected to the jejunostomy tube. The rats were assigned to one of three groups: group 1 received normal saline (control, n = 5); group 2 received EGF at 150 microg/kg/d (n = 5); and group 3 received IGF-1 at 20 mg/kg/d (n = 5). After a 14-day infusion, a portion of mid-small bowel was resected for light and electron microscopic evaluation from each of the animals. The following features were compared between the groups: villous length, crypt length, villous-crypt ratio, villi per millimeter mucosa, goblet cell distribution, eosinophilic infiltrates, number and distribution of organelles, length of microvilli, and completeness of microvillous surface. RESULTS: Ultrastructurally, the bowel epithelium was well preserved in all animals. There were no objective ultrastructural differences between the controls and growth factor-exposed animals. The mean villous-crypt ratio, mean number of villi per millimeter of mucosa (cross section), and mean microvillous height were not significantly different among the groups. However, there was a subjective increase in the number of lysosomes in the enterocytes exposed to EGF and IGF-1. CONCLUSIONS: Administration of EGF and IGF-1 after massive small bowel resection does not appear to significantly alter the small intestine epithelial ultrastructure when compared with the control group. The increase in lysosomes in some of the enterocytes of the animals exposed to growth factors may be important because this finding was not seen in any of the control electron photomicrographs. Studies to evaluate enterocyte gene and protein expression are necessary to determine the mechanism of EGF and IGF-1 enhancement of substrate absorption beyond intestinal adaptation. PMID- 9396550 TI - Excitation-contraction coupling in the day 15 embryonic chick heart with persistent truncus arteriosus. AB - Ca2+ transients were examined in embryonic chick hearts with an experimentally induced cardiac neural crest-related outflow tract defect known as persistent truncus arteriosus (PTA). In all of the animal models of neural crest-related heart defects, prenatal mortality is too high to be attributed to structural defects of the heart alone, suggesting that there is altered development of the myocardium. Earlier reports indicating reduced L-type Ca2+ current in hearts with PTA suggest that poor viability may be related to impairment of cardiac excitation-contraction coupling. To test this hypothesis, direct measurements of the systolic Ca2+ transient in fura-2-loaded myocytes from normal hearts and hearts with PTA were carried out. We found that Ca2+ transients were severely depressed in hearts with PTA and difficult to measure above background noise unless signal averaged or treated with isoproterenol (ISO). We confirmed that the reduced Ca2+ transients were due, at least partly, to a reduction in L-type Ca2+ current. In addition we found that although ISO could raise the L-type current in hearts with PTA to the level found in normal hearts in the absence of ISO, it could not fully restore the Ca2+ transient. Furthermore, caffeine-stimulated Ca2+ transients were diminished in size and the time-to-peak and the decaying phase were significantly slowed. Interestingly, these observations were not accompanied by a reduction in the number of Ca2+ release channels. These results indicated an impairment of SR function in addition to the reduction in L-type Ca2+ current. These results strongly support our hypothesis that the poor viability of embryos with PTA is due to impaired cardiac excitation-contraction coupling. PMID- 9396551 TI - Relative effects of cyclooxygenase and nitric oxide synthase inhibition on vascular resistances in neonatal lamb lungs. AB - Effective attenuation of pulmonary vasoconstriction is essential during early postnatal development when increased pulmonary vascular resistance (PVR) may lead to a resumption of right-to-left shunting across fetal channels. In addition, modulation of venous resistance contributes to normal lung fluid balance. This study was designed to identify the relative modulating effects of endothelium derived nitric oxide (EDNO) and dilator prostaglandins (PG) on normoxic and hypoxic pulmonary vasomotor tone in young newborns. Total and segmental PVR were measured using inflow-outflow and double occlusion techniques in isolated lungs of 6-h-old lambs studied under control conditions or after blocking PG and/or EDNO synthesis with indomethacin and/or N omega-nitro-L-arginine, respectively. During normoxia, both indomethacin and N omega-nitro-L-arginine were required to increase total PVR, but EDNO appeared to have the greater modulating effect. Indomethacin markedly enhanced hypoxic pulmonary vasoconstriction of large and small arteries and small veins, whereas N omega-nitro-L-arginine caused a lesser, but significant, increase in hypoxic pulmonary vasoconstriction of small arteries and veins, suggesting that dilator PG played the dominant modulating role during hypoxia. In addition, PG synthesis appeared to be enhanced after inhibition of EDNO synthesis. In contrast, indomethacin caused a decrease in venous resistance, suggesting that constrictor prostanoids had a greater effect than dilator PG on this segment. EDNO had a modest modulating effect on venous resistance in these lungs. These data suggest that dilator PG and EDNO exert complementary effects in attenuating total and segmental PVR during normoxia and hypoxia in 6-hold lamb lungs. PMID- 9396552 TI - Vascular endothelial growth factor is expressed in ovine pulmonary vascular smooth muscle cells in vitro and regulated by hypoxia and dexamethasone. AB - Chronic lung disease in neonates results from both lung injury and inadequate repair processes. Little is known about the growth factors involved in lung injury and repair, but vascular endothelial growth factor (VEGF) has recently been reported in several animal models of lung injury. VEGF is an endothelial cell-specific mitogen, which is also known as vascular permeability factor because of its ability to induce vascular leak in some tissues. Chronic lung disease is complicated by increased vascular permeability, which can be improved by avoidance of hypoxia and in some cases by dexamethasone therapy. In many cells, hypoxia stimulates VEGF expression. Also, in some cases, dexamethasone blocks VEGF expression. This study examined the role of hypoxia and dexamethasone in regulating the expression of VEGF in pulmonary artery smooth muscle cells. An ovine VEGF cDNA fragment (453 bp) was cloned and found to be highly homologous to known human sequences for VEGF165. Sheep pulmonary artery smooth muscle cells were cultured and exposed to room air, hypoxia, and dexamethasone, alone or in combination for 6 h. At baseline these cells expressed VEGF mRNA at approximately 3.9 kb. The half-life of VEGF mRNA in the smooth muscle cells was 171 min, more than 3-fold longer than previous reports for epithelial cells. Exposure to hypoxia caused a 3-fold increase in mRNA abundance, primarily through transcriptional up-regulation. Dexamethasone blocked the hypoxia-induced increase in VEGF mRNA. The results demonstrate that hypoxia and dexamethasone are regulators of VEGF expression in ovine pulmonary artery smooth muscle cells. It is not known whether VEGF derived from these cells is involved in lung injury and/or normal homeostatsis. PMID- 9396553 TI - Developmental changes in the effect of acidosis on contraction, intracellular pH, and calcium in the rabbit mesenteric small artery. AB - The purpose of the present study was to determine developmental changes in the effect of respiratory acidosis on vascular smooth muscle contraction. Vessel diameter, intracellular pH (pHi), and calcium concentration ([Ca]i) were measured in a cannulated preparation of the small mesenteric artery of newborn and adult rabbits. In the artery precontracted by high KCl, acidosis caused a vasorelaxation both in the newborn and the adult; the vasorelaxation was greater in the newborn than in the adult. The fura-2 fluorescence ratio, an indicator of [Ca]i, decreased transiently during acidosis and the decrease was similar in the two age groups. In the artery precontracted by norepinephrine, acidosis caused a transient vasoconstriction in the adult and a vasorelaxation in the newborn. In these vessels, the fura-2 fluorescence ratio increased transiently during acidosis; the increase was similar in the two groups. Upon induction of acidosis, pHi fell rapidly in the artery precontracted by norepinephrine or high KCl, and the depression of pHi was similar in the two groups. In the skinned smooth muscle preparation, a tension-[Ca] relationship curve at pH 7.1 was not significantly different from that at pH 6.8 in the adult. In the newborn, the tension-[Ca] curve at pH 6.8 was shifted to the right, compared with that at pH 7.1. These data suggest that the vasorelaxant effect of respiratory acidosis in the premature vessel is greater than in the adult. The greater vasorelaxation in the newborn cannot be explained by the age-related difference in pHi or [Ca]i during acidosis. The greater sensitivity of myofibrils to low pHi in the newborn may, at least in part, be responsible for the greater vasorelaxation in this age group. PMID- 9396554 TI - Perinatal growth disturbance in the spontaneously hypertensive rat. AB - Disproportionate fetal and placental growth are associated with the development of hypertension in the rat and human. Here we report differences in fetal, neonatal, and placental growth, and in metabolism and endocrinology, between the spontaneously hypertensive rat (SHR), a genetic model for human essential hypertension, and the control Wistar-Kyoto (WKY) strain. Gestation in SHR (23 d) was longer than in WKY by 20 h. Body weights were lower in the SHR from fetal d 16 to 20 and on postnatal d 15. However, on fetal d 22 and postnatal d 1, there was no significant difference in body weight between SHR and WKY. SHR placentas were larger than those of WKY at d 20, and by term there was a difference of 30% (p < 0.01). Other indices of disproportionate growth were hypertrophy of the fetal heart and kidney and decreased ponderal index in the SHR neonate. Blood glucose in SHR fetuses was lower than in WKY fetuses (p < 0.05), whereas blood lactate was higher (p < 0.05) and fetal hematocrit was reduced (p < 0.001). These findings suggest undernutrition and placental insufficiency may occur in SHR fetuses. Plasma IGF-II was increased on the last day of gestation in both strains, whereas IGF-I was unaltered. Fetal liver IGFBP-2 mRNA and plasma IGFBP-2 levels were reduced in SHR on fetal d 20 and 22 (p < 0.01). Differences in growth and endocrine and metabolic parameters suggest abnormal perinatal physiology in the SHR, which may influence the later development of hypertension. PMID- 9396555 TI - Fetal atrioventricular, venous, and arterial flow velocity waveforms in the small for gestational age fetus. AB - Arterial, venous, and intracardiac Doppler flow velocity waveforms were studied in 50 women with a small for gestational age (SGA) fetus according to a cross sectional study design. No Doppler signals could be obtained in five women for technical reasons. The remaining 45 women were compared with normal control subjects matched for gestational age and maternal parity. The 45 SGA fetuses were divided into birth weight below the 5th centile for gestational age (group I, n = 35) and birth weight between the 5th and 10th centile for gestational age (group II, n = 10). A significant difference in baseline characteristics was found between both SGA subsets and normal controls. In SGA I fetuses, the pulsatility index in the umbilical artery and descending aorta was significantly higher, but lower in the middle cerebral artery when compared with normal controls. At the atrioventricular and venous level (umbilical vein, ductus venosus, and inferior vena cava) reduced time-averaged velocities were established. PIV in the ductus venosus and IVC showed a significant increase. Within the same SGA subset, no relationship could be established between arterial downstream impedance and 1) atrioventricular flow velocities and 2) pulsatility index in the ductus venosus and inferior vena cava. Also, no relationship existed between flow velocity waveforms and pregnancy-induced hypertension and admission to the neonatal intensive care unit. Umbilical venous pulsations and absent/reverse flow in the umbilical artery were associated with a high intrauterine mortality rate and low birth weights. In SGA II fetuses, the pulsatility index in the umbilical artery and descending aorta was significantly higher than in normal controls. It can be concluded that fetuses with a birth weight below the 5th centile demonstrate marked changes in arterial, atrioventricular, and venous flow velocity waveforms. Atrioventricular and venous flow velocity waveforms change independently from arterial downstream impedance, suggesting that other factors, such as reduced volume flow and myocardial contraction force, may play a role in the observed changes. PMID- 9396557 TI - The role of recombinant platelet-activating factor acetylhydrolase in a neonatal rat model of necrotizing enterocolitis. AB - Previous studies have shown that the endogenous inflammatory mediator platelet activating factor (PAF) plays an important role in the pathophysiology of neonatal necrotizing enterocolitis (NEC). This study was designed to investigate the role of the PAF-degrading enzyme acetylhydrolase (PAF-AH) in a neonatal rat model of NEC. To study the absorption, localization, and activity of human recombinant PAF-AH (rPAF-AH), newborn rats were treated with enteral rPAF-AH, and plasma and intestines were sampled at 8 and 24 h for determination of PAF-AH enzyme activity and rPAF-AH concentration using a specific enzyme-linked immunoassay. To study the effect of rPAF-AH on neonatal NEC, rats were treated with rPAF-AH via the enteral route every 3 h, and then subjected to formula feeding and asphyxia per an established neonatal rat protocol for NEC. Pretreatment with enteral rPAF-AH significantly reduced the incidence of NEC compared with controls (6/26 versus 19/26, p < 0.001). We found that enteral rPAF AH administration resulted in significant intestinal PAF-AH activity but no circulating PAF-AH activity despite immunohistochemical localization of the administered rPAF-AH to the intestinal epithelial cells. These findings suggest that rPAF-AH is functional and stable in the gut of neonatal rats. We conclude that enteral administration of rPAF-AH remains locally active and reduces the incidence of NEC in our experimental animal model. PMID- 9396556 TI - Correlation between the functional assay for activated protein C resistance and factor V Leiden in the neonate. AB - A factor V506 Arg-Gln mutation is the most common inherited cause of thrombophilia in adults. To date, there are no data regarding the detection of this mutation in neonatal blood or the relationship of this dysfunctional factor V to neonatal thrombosis. This study compared a modified activated protein C resistance functional assay with the PCR-based DNA assay for the factor V mutation in 115 prospectively collected umbilical cord blood samples. The incidence of activated protein C resistance in cord blood was 6%. The sensitivity and specificity of the modified assay for the factor V Leiden mutation was 100%. PMID- 9396558 TI - Hepatitis G virus infection in normal and prospectively followed posttransfusion children. AB - A recently identified RNA virus, hepatitis G virus (HGV), has been investigated for its role in causing non-A-E hepatitis. The frequency and clinical outcome of HGV infection in children was studied. Two hundred apparently healthy children aged 6 mo to 12 y, and 90 children who had undergone open heart surgery in a prospective study for posttransfusion hepatitis were included in this study. The serum samples were tested for HGV RNA by nested reverse transcription-PCR with primers from the 5'-untranslated region. The HGV RNA viremic rate was found to be 1% (2/200) in apparently healthy children, 30% in children after open heart surgery. Among the 90 children, three were HGV-infected before the surgery. Twenty-four (28%) of the remaining 87 children tested positive for HGV RNA within 6 mo after the surgery. Sixty-five percents of these viremic children eventually became persistently infected at 1 y after surgery. No HGV RNA-positive children exhibited elevated alanine aminotransferase levels during the follow-up period. No coinfections of HGV with the hepatitis C virus or hepatitis B virus were found. Patients of younger age appeared more likely to become chronic carriers. Anti-HCV screening did not reduce the prevalence of HGV infection. In conclusion, in children with open heart surgery, the risk of transfusion-transmitted HGV infection and the chronicity rate have been found to be high. Young age is a risk factor of persistent infection. PMID- 9396559 TI - The production of macrophage inflammatory protein-1alpha in the cerebrospinal fluid at the initial stage of meningitis in children. AB - Neutrophils in the cerebrospinal fluid (CSF) increase during the initial stage of meningitis. Some cytokines induce the accumulation of such neutrophils, and we and other investigators have revealed transient increases in the levels of granulocyte-colony stimulating factor (G-csf) and IL-8 in the CSF of patients with meningitis. To explore the coordination of other cytokines with G-csf and IL 8 in the neutrophil accumulation in the CSF, we herein investigated macrophage inflammatory protein-1alpha (MIP-1alpha), which can induce the infiltration of neutrophils. The modulation of MIP-1alpha levels in the CSF in children with bacterial (n = 10) and aseptic (n = 22) meningitis was examined using an ELISA. MIP-1alpha levels in the CSF were detectable at the stage with symptoms of meningitis: 289.9 +/- 270.7 ng/L in the bacterial meningitis group and 16.1 +/- 12.5 ng/L in the aseptic meningitis group. These levels decreased with the improvement of symptoms. MIP-1alpha was not detectable (<6 ng/L) in all of the control patients without meningitis (n = 19). The MIP-1alpha levels in the CSF showed a significant correlation with the CSF neutrophil counts (r = 0.750, p < 0.0001; n = 80) of meningitis, and the values of MIP-1alpha (log ng/L)/neutrophil counts (log/L) ratio were calculated (1.003 +/- 0.576). The MIP-1alpha levels in the serum were significantly lower than those in the CSF (p = 0.0464). We found MIP-1alpha mRNA in the CSF cells by the reverse transcriptase-PCR method, and high levels of MIP-1alpha protein in the culture media from mononuclear cells in the CSF in vitro. In summary, The MIP-1alpha level increases in the CSF at the symptomatic stage of meningitis in children, and its cellular source is, in part, mononuclear cells which have infiltrated the CSF. We propose that MIP-1alpha, in addition to G-csf and IL-8, plays an important role in the accumulation of neutrophils in the CSF of patients with meningitis. PMID- 9396560 TI - Immune abnormalities in guinea pigs with asymptomatic congenital syphilis. AB - Spleens from 1-20-wk-old guinea pigs infected in utero with Treponema pallidum and age-matched controls, born to normal and heat-killed (56 degrees C, 2 h.) T. pallidum-injected mothers, were examined for their in vitro lymphoproliferative response to phytohemagglutinin, concanavalin A, and lipopolysaccharide. Additionally, T cell surface markers (mu-chain, pan T, CD4, and CD8) were determined in spleen, lymph node, and peripheral blood from 10-wk infected and normal pups by single and dual parameter fluorescence-activated cell sorter analysis. Compared with control animals, congenitally infected animals showed a remarkable prolonged naive-type of immune response as reflected by the higher (p < 0.01) proliferative responses to both T cell mitogens (up to 20 wk of age), and the weaker response to the B cell mitogen, significantly different (p < 0.01) at 10 wk of age. As opposed to controls, in all organs examined the level of CD8+ (cytotoxic/suppressor) T cells was significantly diminished (p < 0.01); consequently, the CD4/CD8 ratio was significantly elevated (p < 0.05). The role of C4 complement component and the nature and potential role of the immature T and B lymphocyte responses in asymptomatic congenital syphilis is discussed. PMID- 9396561 TI - Penetration of group B streptococci through polarized Madin-Darby canine kidney cells. AB - Group B streptococci (GBS) are one of the major causes of invasive neonatal infection. The pathogenesis of early onset disease is a multistep process. Adhesion of GBS to eucaryotic cells is considered to be an important step for the establishment of infection. Subsequent to adhesion, GBS invade cells and give rise to septicemia and meningitis. To investigate passage of GBS across epithelial cell linings we examined the interaction between bacteria and Madin Darby canine kidney (MDCK) cells. When grown on permeable support, these cells form a polarized epithelial monolayer with an apical-to-basolateral orientation, which more reflects the in vivo situation compared with conventionally cultured cells. Our results show that GBS are translocated in vacuoles from the apical to the basolateral surface of MDCK cells in a temperature-dependent process. The passage of GBS through the cells is selective with only small numbers of bacteria penetrating in the basolateral-to-apical direction. Transcytosis of GBS starts before decrease in transepithelial resistance of the monolayer. These data suggest a mechanism for traversal of GBS over intact chorioamniotic membranes and from alveoli into the circulation of the fetus. PMID- 9396562 TI - Impact of placental restriction on the development of the sympathoadrenal system. AB - We have investigated the impact of chronic restriction of placental function on circulating catecholamine concentrations and responses to the indirectly acting, sympathomimetic amine, tyramine, in the fetal sheep in late gestation. In 10 ewes, endometrial caruncles or placental placentation sites were removed before conception (placental restriction (PR) group). Fetal sheep in the PR group were hypoxemic throughout late gestation and growth-restricted (3.02 +/- 0.35 kg) when compared with control fetal sheep (4.30 +/- 0.29 kg; n = 8) at 140 d of gestation. Fetal plasma concentrations of noradrenaline and adrenaline were higher (p < 0.05) in the PR (7.06 +/- 3.17 pmol/mL and 2.89 +/- 2.01 pmol/mL, respectively) than in the control group (3.55 +/- 0.54 pmol/mL and 1.30 +/- 0.48 pmol/mL, respectively) throughout late gestation. Plasma noradrenaline, but not adrenaline concentrations, increased significantly between 110 and 140 d of gestation in both the PR and control group, and there was a significant inverse relationship between plasma noradrenaline and arterial PO2 in the PR and control groups (plasma noradrenaline = 12.34 - 0.40 PO2). In the PR group, plasma noradrenaline increased (p < 0.05) after tyramine infusion from 4.51 +/- 1.28 pmol/mL to a peak of 19.40 +/- 3.56 pmol/mL. In the control group, noradrenaline increased from 2.08 +/- 0.30 pmol/mL to a peak of 12.23 +/- 1.67 pmol/mL after tyramine infusion. There was no difference, however, in the maximal proportional changes in plasma noradrenaline concentrations in the PR (319 +/- 55%) and control (449 +/- 100%) groups after tyramine. We conclude that the most likely source of the increased plasma catecholamines in the PR group is enhanced catecholamine synthesis and secretion from developing sympathetic neurons. PMID- 9396563 TI - Tissue metabolism and plasma levels of thyroid hormones in critically ill very premature infants. AB - Thyroid status was characterized in very preterm infants (gestational age < or =32 wk; n = 61) from birth through d 14, and in infants who died within 16 d after delivery (n = 10), where it was also correlated with metabolism of iodothyronines in peripheral tissues (brain, liver, kidney, skeletal muscle, and adipose tissue). At 3 d of life, mean plasma levels of thyroxine, triiodothyronine, and TSH started to decrease, being lower in the critically ill compared with healthy premature neonates. Activities of the three iodothyronine deiodinases enzymes (type I, II, and III, respectively) were detected in all postmortem tissue samples, except for absence of the type II activity in kidney. All activities were the highest in liver and differed in other tissues. Lack of correlation between the type I activity in liver (and kidney), and plasma levels of thyroid hormones suggested that the thyroid was the primary source of circulating triiodothyronine. On the other hand, namely in brain, correlations between activity of the deiodinases and plasma hormone levels were found which suggested a complex control by thyroid hormones of their own metabolism. High activity of type III in liver, adipose tissue, and skeletal muscle demonstrated a role of these tissues in thyroid hormones degradation. Results support the view that peripheral tissues of very preterm infants are engaged in local generation of triiodothyronine, and inactivation of thyroid hormones, but do not represent a major source of circulating triiodothyronine. PMID- 9396564 TI - Assessment of the efficacious dose of arachidonic and docosahexaenoic acids in preterm infant formulas: fatty acid composition of erythrocyte membrane lipids. AB - The nutritional requirements of preterm infants for the long chain polyunsaturated essential fatty acids, arachidonic acid (AA) and docosahexaenoic acid (DHA), have not been clearly defined. The present study evaluated preterm infants of less than 2.3 kg birth weight fed a commercial formula (Preemie SMA) devoid of AA and DHA and compared this control group with similar infant groups fed one of three formulas containing a range of 0.32 to 1.1% AA and 0.24 to 0.76% DHA. An analogous group of infants fed their mothers' breast milk and a breast milk fortifier (when indicated) was also studied. Erythrocyte membrane phospholipids were isolated from blood samples collected at 12 d of age and after a further 4 wk of feeding. Infants fed the formula without AA and DHA showed a reduction in AA level in erythrocyte phosphatidylcholine, and a reduced level of DHA in phosphatidylethanolamine in comparison with infants fed breast milk or infant formula containing AA and DHA. Supplementing infant formula with increasing levels of AA and DHA produced a clear dose response in the levels of AA and DHA found in erythrocyte membrane phospholipids. From comparison of membrane phospholipid fatty acid composition it appears that a formula level of 0.32-1.1% AA and 0.24-0.76% DHA provides sufficient levels of these fatty acids to achieve a similar fatty acid composition to that of infants fed human milk for most of the lipid fractions examined. PMID- 9396565 TI - Brain docosahexaenoate accretion in fetal baboons: bioequivalence of dietary alpha-linolenic and docosahexaenoic acids. AB - The dietary bioequivalence during the brain growth spurt of alpha-linolenic (LNA) and docosahexaenoic acids (DHA) as substrates for brain and retinal n-3 fatty acid accretion is reported for the fetal baboons, whose mothers consumed a long chain polyunsaturate-free diet with a n-6/n-3 ratio of 10:1. Pregnant baboons received i.v. doses of U-13C-labeled fatty acids (LNA or DHA), plasma was collected from mother and fetus, and fetal brain (occipital cortex), retina, and liver were analyzed at various times post-dose. Fetal brain DHA plateaued 15-35 d post-dose with 1.6% of the preformed [U-13C-]DHA dose recovered in the brain. In contrast, LNA-derived DHA accretion also plateaued but was 20-fold lower. Liver and retinal results were of the same order of magnitude, but showed evidence of peaks and decline. Conversion products to n-3 long chain polyunsaturate were observed in the maternal circulation at 1 h after administration, as was transfer of both fatty acids to the fetus. From these measurements we estimate that a dietary level of about 0.45% of energy as LNA is sufficient to meet the requirements of the growing fetal brain, whereas 0.03% of energy as DHA would suffice. These data are the first direct measurements of the bioequivalence of DHA and LNA in developing primates and imply that n-3 fatty acid requirements for the developing fetal brain can be met by attainable dietary LNA for diets low in long chain polyunsaturates. PMID- 9396566 TI - Reducing cell membrane n-6 fatty acids attenuate mucosal damage in food-sensitive enteropathy in mice. AB - Mucosal damage is commonly observed in food-sensitive enteropathy in infants, and the generation of leukotrienes is involved in the pathogenesis of this enteropathy. Because supplementing n-3 fatty acids is known to modify the production of leukotrienes, we investigated whether a change of dietary fatty acid composition affects leukotriene synthesis and food hypersensitivity reactions in the intestine by using a mouse model of food-sensitive enteropathy. The model was prepared by feeding ovalbumin to BALB/c mice after intraperitoneal injection of cyclophosphamide. Diets were prepared from soybean oil (control), perilla oil, lard, corn oil, and 0.125 volume of corn oil (low fat diet) and given to mice for 4 wk. Villous heights, crypt depths, leukotriene B4 and C4 production in the intestine were measured. Crypt hyperplasia and villous atrophy were severer in the corn oil-fed group than those of control group, whereas mucosal damage in the perilla oil and low fat diet groups was minimal. In the corn oil-fed group, red blood cell membrane levels of n-3 fatty acids were lower than the control, and the synthesis of leukotrienes was highest among all groups. In the perilla oil and low fat diet groups, n-6 fatty acids were lower than those of control group and leukotriene production was significantly suppressed. These results indicate that reducing cell membrane levels of n-6 fatty acids by feeding less n-6 fatty acids or supplementing n-3 fatty acids, is important to suppress leukotriene biosynthesis for prevention from mucosal damage in food-sensitive enteropathy. PMID- 9396567 TI - Mutations in the human biotinidase gene that cause profound biotinidase deficiency in symptomatic children: molecular, biochemical, and clinical analysis. AB - Biotinidase deficiency is an autosomal recessively inherited disorder that results in the inability to recycle the vitamin biotin. The disorder can cause neurologic and cutaneous abnormalities that can be treated effectively with pharmacologic doses of biotin. We identified 21 mutations that cause profound biotinidase deficiency in 37 symptomatic children (30 different probands and 7 siblings), as well as provide relevant biochemical and clinical information for each child. The two most common mutations (G98:d7i3 and R538C) were found in 31 of 60 alleles (52%), whereas the remainder of the alleles are accounted for by the 19 other unique mutations. Serum samples were available from 18 children, of these 11 had no detectable cross-reacting material (CRM) to antibody prepared against normal human serum biotinidase, three had reduced quantities of CRM and four had normal quantities of CRM in serum. All of these mutations result in complete absence of biotinyl-transferase activity in serum. Two polymorphisms were also identified in normal individuals. It is apparent that a child who inherits any of these mutations, either in the homozygous state or in combination, can develop the clinical features of the disorder if untreated. There are, however, no clear genotype/phenotype correlations that would allow for the prediction of the type, severity, or age of onset of symptoms. PMID- 9396568 TI - Characterization of mutant holocarboxylase synthetase (HCS): a Km for biotin was not elevated in a patient with HCS deficiency. AB - Holocarboxylase synthetase (HCS) is an essential enzyme for the biotinylation of several mammalian carboxylases. A deficiency of HCS is accountable for early onset biotin-responsive multiple carboxylase deficiency. To address the mechanism of biotin responsiveness, we analyzed the kinetic properties of the previously identified mutant, L237P, and another mutant, V550M, described in this report. The V550M mutant contains a G to A transition at position 1935, which is within the putative biotin binding site, whereas the mutation in L237P occurs outside the biotin binding site. Km and Vmax values for the mutant proteins were determined by overexpressing cDNAs encoding the mutants in transformed fibroblasts from an HCS-deficient patient. Enzyme activity assays were performed using apo-carboxyl carrier protein as a substrate. A Km for biotin that was larger than the value found for the wild-type cDNA was observed in fibroblasts transfected with the V550M cDNA, but not the L237P cDNA. The Vmax for the expressed L237P cDNA was 4.3% of that observed for the wild-type cDNA. Biotin responsiveness in the patient with the L237P mutation was neither due to an increased affinity for biotin nor a restoration of stability of the mutant by biotin treatment. A new mechanism of biotin responsiveness in HCS deficiency is presented. PMID- 9396569 TI - Urinary galactonate in patients with galactosemia: quantitation by nuclear magnetic resonance spectroscopy. AB - Although numerous reports have appeared showing high levels of galactitol in the urine of patients with galactose-1-phosphate uridylyltransferase deficiency, little attention has been paid to measurement of urinary galactonate. Herein we explored the use of 1H and 13C nuclear magnetic resonance, which required only the concentration of urine without derivatization, to detect and quantitate urinary galactonate. We report that transferase deficient infants, as well as adults on galactose restricted diets excrete significant amounts of galactonate, whereas none is detected in the urine of normal subjects. Galactose-toxic infants were found to excrete large amounts of galactonate, which decreased when the lactose-free diet was instituted. We also found that normal individuals subjected to an oral galactose load also excrete high levels of galactonate for at least 4 h after galactose ingestion. Our data provide evidence that the first reaction in the oxidative pathway of galactose metabolism described in rat liver in 1966 is activated in patients with a variety of galactose-1-phosphate uridylyltransferase gene mutations even while on a lactose-restricted diet. In both patients and normal individuals, flux through the alternate galactonate pathway appears to be related to the body galactose burden. PMID- 9396570 TI - In vitro copper stimulation of plasma peptidylglycine alpha-amidating monooxygenase in Menkes disease variant with occipital horns. AB - We determined the concentrations of copper, the activities of ceruloplasmin and peptidylglycine alpha-amidating monooxygenase (PAM), and the stimulation index of PAM by the in vitro addition of copper in plasma samples obtained from three male patients with occipital horns and a milder Menkes disease phenotype, having severe copper deficiency due to the defect in copper transport. We found a decreased plasma ceruloplasmin activity and an increased copper stimulation index of plasma PAM in these patients compared with healthy control subjects. The combination of these two determinations may provide a means for the assessment of copper nutriture in humans using blood samples obtained in a single microhematocrit tube. Further investigation is warranted to evaluate whether these noninvasive measurements can be used for the diagnosis of mild copper deficiency in humans with sufficient specificity and sensitivity. PMID- 9396571 TI - Deficit of brain and skeletal muscle bioenergetics and low brain magnesium in juvenile migraine: an in vivo 31P magnetic resonance spectroscopy interictal study. AB - We used phosphorus magnetic resonance spectroscopy (31P MRS) to investigate in vivo the brain and skeletal muscle energy metabolism of 15 children with migraine with aura in interictal periods. Brain 31P MRS disclosed low phosphocreatine and high inorganic phosphate contents, and high intracellular pH in all patients. Calculated [ADP] and the relative rate of mitochondrial oxidation were higher in the brain of patients than in control subjects, whereas the phosphorylation potential was lower. Brain intracellular free Mg2+ concentration was reduced by 25% in patients. Abnormal skeletal muscle mitochondrial respiration was also disclosed in 7 of 15 patients as shown by the slow rate of phosphocreatine postexercise recovery. The multisystem bioenergetic failure found in patients with juvenile migraine is comparable to that found in adults with different types of migraine. PMID- 9396572 TI - Maturation of anoxia-induced gasping in the rat: potential role for N-methyl-D aspartate glutamate receptors. AB - After anoxia-induced apnea, gasping remains the last operative mechanism for survival. In developing rats, the gasping response to anoxia exhibits triphasic characteristics. Because anoxia is associated with enhanced release of glutamate, we hypothesized that N-methyl-D-aspartate (NMDA) glutamate receptors may underlie components of the gasping response. Rat pups aged 2 d (n = 50), 5 d (n = 43), 10 d (n = 42), and 15 d (n = 45) underwent anoxic challenges with 100% N2 in a whole body plethysmograph, 30 min after intraperitoneal administration of MK801 [(+)-5 methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine hydrogen maleate; dizocilpine] (3 mg/kg), a noncompetitive NMDA glutamate receptor channel antagonist, or normal saline. In control pups, after primary apnea onset, a triphasic gasping pattern was apparent at all postnatal ages and included two distinct types of gasps (I and II). In 2- and 5-d MK801-treated animals, phase 1 and type I gasps were absent, leading to marked prolongations of the gasp latency and phase 2, the latter displaying type II gasps only. In addition, phase 3 duration was also prolonged with increased type II gasp frequencies. In contrast, in some 10-d-old (40%) and in all 15-d-old MK801-treated pups, although overall gasping duration was prolonged, the triphasic gasping pattern seen in matched controls was also present. We conclude that NMDA glutamate receptors mediate particular phasic components of the gasping response during early postnatal life but not at later stages of development. We speculate that developmental changes occur in both function and expression of NMDA and other neurotransmitters within brainstem regions underlying the neural substrate for gasp generation. PMID- 9396573 TI - Mixed venous oxygen saturation and biochemical parameters of hypoxia during progressive hypoxemia in 10- to 14-day-old piglets. AB - In this study we wanted to assess the relationship between mixed venous oxygen saturation (SVO2) and tissue oxygenation. For that, we compared the values of SVO2 with oxygen delivery (DO2), oxygen consumption (VO2), and markers of tissue hypoxia such as lactate and pyruvate during progressive hypoxemia. Eight 10-14-d old piglets were anesthetized, tracheotomized, intubated, and ventilated. A fiberoptic catheter was placed in the carotid artery to monitor arterial oxygen saturation (SaO2). A thoracotomy was performed, and a fiberoptic catheter was placed in the pulmonary artery to monitor SVO2. A transit time ultrasound flow probe was positioned around the ascending aorta to measure aorta flow. Progressive graded hypoxemia was induced by decreasing fractional inspiratory oxygen concentration (FIO2) from 1.0 to 0.30, 0.21, 0.15, and 0.10. After each FIO2 interval blood samples were taken for blood gases, lactate, and pyruvate. DO2 and VO2 were calculated. SVO2 decreased similarly to SaO2. A value of SVO2 of more than 40% excluded oxygen restricted metabolism. When DO2 decreased below a critical range (8.4-12.8 mL/kg x min), SVO2 decreased below 15%, and lactate and the lactate/pyruvate ratio increased. We conclude 1) that baseline SVO2 values excluded oxygen-restricted metabolism, 2) that SVO2 values between 15 and 40% were not a marker for oxygen-restricted metabolism, and 3) that SVO2 values below 15% were associated with oxygen-restricted metabolism. Reduced SVO2 values must be interpreted as a change of the factors that determine the balance between DO2 and VO2 and as a warning that, with further reduction of SVO2, oxygen restricted metabolism can develop. PMID- 9396574 TI - Postnatal lung function after prenatal steroid treatment in sheep: effect of gender. AB - The effect of fetal gender on postnatal lung function and response to prenatal steroid exposure were examined retrospectively in a group of 115 preterm lambs. Fetuses received a single intramuscular injection of 0.5 mg/kg betamethasone alone or in conjunction with L-thyroxine 48 h before delivery at 128-d gestational age. Control animals received an equivalent volume of saline. After delivery, respiratory mechanics and blood gas parameters were recorded for 40 min. Deflation pressure volume curves were constructed in excised lungs. Right upper lobes from a randomly selected subgroup of control animals were examined morphometrically. Control (saline-treated) females were able to be ventilated at lower ventilatory pressures with equivalent tidal volumes and more efficient gas exchange. There were no gender differences in compliance, conductance, or excised lung volumes for saline-treated animals. More efficient gas exchange in females could not be explained by thinner alveolar septa or greater alveolar surface area. After hormone treatment, both males and females exhibited significant improvements in respiratory mechanics, gas exchange, and an increase in alveolar surfactant concentration. However, female exhibited a significantly greater improvement than males for compliance, conductance, excised lung volume, and arterial oxygen partial pressure. These data provide a comprehensive description of gender differences in postnatal lung function and response to steroid treatment in preterm animals, and support clinical findings of sexual dimorphism. PMID- 9396575 TI - Repeated doses of the perfluorocarbon FC-100 improve lung function of preterm lambs. AB - Intratracheal administration of a single dose of the perfluorocarbon FC-100 improves lung function in surfactant-deficient animals. In this study we compared the response to repeated doses of FC-100 (3 mL/kg 3% solution, n = 5) with that observed after administration of Exosurf (5 mL/kg, n = 5) to mechanically ventilated preterm lambs of 125 d of gestation. The initial dose of FC-100 rapidly increased arterial PO2, decreased arterial PCO2, and improved arterial pH. Also dynamic lung compliance markedly improved with this agent. Administration of an additional dose of FC-100 resulted in relatively similar changes, albeit of lesser magnitude than those observed with the initial dose. In contrast, Exosurf did not improve these variables even after three doses. All lambs treated with FC-100 survived the 6-h study period, whereas one of the five Exosurf-treated lambs survived (p < 0.05). Mean arterial blood pressure and heart rate decreased in those lambs that received FC-100, but not in surviving lambs that received Exosurf. Our data demonstrate that repeated intratracheal administration of the perfluorocarbon FC-100 improves lung function and survival of surfactant-deficient lambs better than the synthetic surfactant Exosurf. We speculate that tensio-active agents with properties different from surfactant, such as FC-100, might improve lung function in preterm neonates with diseases due to surfactant deficiency. PMID- 9396576 TI - Low birth weight and subsequent male subfertility. AB - Male subfertility often remains unexplained. Severe intrauterine growth retardation has previously been linked to hypergonadotropic hypogonadism. We examined whether reduced fetal growth, as judged by low birth weight, is associated with unexplained male subfertility later in life. Birth weight and gestational age were obtained by questionnaire from male partners of couples consulting for subfertility, and were transformed into birth weight SD scores. Men with normal semen analysis (n = 128) had a median birth weight SD score of 0.0 (P25-P75 range: -0.7 to 1.0), comparable to that of men with explained subfertility (n = 28), and higher (p = 0.012) than that of men with unexplained subfertility (n = 32; median -0.5 SD score; P25-P75 range: -0.9 to -0.1). These results extend the link between reduced fetal growth and male subfertility to a range of birth weight that is well within normality. The pathophysiologic mechanism governing this association now remains to be unraveled. PMID- 9396577 TI - American Pediatric Society John Howland Award 1997: presentation and acceptance. PMID- 9396578 TI - American Pediatric Society Presidential Address 1997: the ever-whirling wheel of change. PMID- 9396579 TI - Society for Pediatric Research Presidential Address 1997: multiculturalism in research. PMID- 9396580 TI - Editors and ethics. PMID- 9396581 TI - Natural painkillers. PMID- 9396582 TI - Effects of RU486 on HIV-1 replication. PMID- 9396583 TI - Black market drugs are only the tip of the iceberg. PMID- 9396584 TI - The most dangerous woman in the world? PMID- 9396585 TI - Does Sampson see any alternative? PMID- 9396586 TI - NIH says "yes" to acupuncture. PMID- 9396587 TI - UNAIDS expands HIV drug access in developing countries. Joint United Nations Programme on HIV/AIDS. PMID- 9396588 TI - Bill may double funds for NIH bone research. PMID- 9396589 TI - Japan to legalize the pill. PMID- 9396590 TI - Reducing embryo implants in IVF. PMID- 9396591 TI - Kornberg calls for scientists' association. PMID- 9396592 TI - Medical research: the national interest. PMID- 9396593 TI - Plasmodium falciparum malaria--sticky jams and PECAM pie. PMID- 9396594 TI - Cystic fibrosis lung infection cleared up? PMID- 9396595 TI - The multi-faceted personality of HIV. PMID- 9396596 TI - Rats go with the (urine) flow. PMID- 9396597 TI - More mayhem from molecular mimics. PMID- 9396598 TI - Puzzling over prion partners. PMID- 9396599 TI - Act locally, think globally. PMID- 9396600 TI - Rotavirus vaccines at the threshold. PMID- 9396601 TI - Prostaglandins and reproduction--what do knockouts really tell us? PMID- 9396602 TI - Regeneration in the adult central nervous system: experimental repair strategies. PMID- 9396603 TI - Dye efflux studies suggest that hematopoietic stem cells expressing low or undetectable levels of CD34 antigen exist in multiple species. AB - We previously described a method for isolating murine hematopoietic stem cells capable of reconstituting lethally irradiated recipients, which depends solely on dual-wavelength flow cytometric analysis of murine bone marrow cells stained with the fluorescent DNA-binding dye Hoechst 33342. This method, which appears to rely on the differential ability of stem cells to efflux the Hoechst dye, defines an extremely small and homogeneous population of cells (termed SP cells). We show here that dual-wavelength analysis of Hoechst dye-stained human, rhesus and miniature swine bone marrow cells reveals a small, distinct population of cells that efflux the dye in a manner identical to murine SP cells. Like the murine SP cells, both human and rhesus SP cells are primarily CD34-negative and lineage marker-negative. In vitro culture studies demonstrated that rhesus SP cells are highly enriched for long-term culture-initiating cells (LTC-ICs), an indicator of primitive hematopoietic cells, and have the capacity for differentiation into T cells. Although rhesus SP cells do not initially possess any hematopoietic colony forming capability, they acquire the ability to form colonies after long-term culture on bone marrow stroma, coincident with their conversion to a CD34 positive phenotype. These studies suggest the existence of a hitherto unrecognized population of hematopoietic stem cells that lack the CD34 surface marker classically associated with primitive hematopoietic cells. PMID- 9396604 TI - Murine gamma-herpesvirus 68 causes severe large-vessel arteritis in mice lacking interferon-gamma responsiveness: a new model for virus-induced vascular disease. AB - Fundamental issues remain unresolved regarding the possible contribution of viruses to vascular pathology, as well as the role of the immune system in regulating these processes. Here we demonstrate that infection of mice with gamma herpesvirus 68 (gammaHV68) provides a novel model for addressing these issues. Interferon-gamma receptor-deficient (IFNgammaR-/-) mice died weeks to months after gammaHV68 infection from a severe large-vessel panarteritis. GammaHV68 infected B cell-deficient and normal weanling mice exhibited milder large-vessel arteritis. Immunohistochemical analyses demonstrated gammaHV68 antigen in arteritic lesions and revealed a striking tropism of gammaHV68 for smooth muscle cells. These studies demonstrate that IFN-gamma is essential for control of chronic vascular pathology induced by gammaHV68 and suggest gamma-herpesviruses as candidate etiologic agents for human vasculitis. PMID- 9396605 TI - Therapy of malignant brain tumors by intratumoral implantation of retroviral vector-producing cells. AB - Intratumoral implantation of murine cells modified to produce retroviral vectors containing the herpes simplex virus-thymidine kinase (HSV-TK) gene induces regression of experimental brain tumors in rodents after ganciclovir (GCV) administration. We evaluated this approach in 15 patients with progressive growth of recurrent malignant brain tumors. Antitumor activity was detected in five of the smaller tumors (1.4 +/- 0.5 ml). In situ hybridization for HSV-TK demonstrated survival of vector-producing cells (VPCs) at 7 days but indicated limited gene transfer to tumors, suggesting that indirect, "bystander," mechanisms provide local antitumor activity in human tumors. However, the response of only very small tumors in which a high density of vector-producing cells had been placed suggests that techniques to improve delivery and distribution of the therapeutic gene will need to be developed if clinical utility is to be achieved with this approach. PMID- 9396606 TI - Tumor regression with regional distribution of the targeted toxin TF-CRM107 in patients with malignant brain tumors. AB - We investigated regional therapy of recurrent malignant brain tumors with transferrin-CRM107, a conjugate of human transferrin (Tf) and a genetic mutant of diphtheria toxin (CRM107) that lacks native toxin binding. Physiological barriers to delivering proteins to tumor and surrounding infiltrated brain were circumvented with high-flow interstitial microinfusion. At least a 50% reduction in tumor volume on magnetic resonance imaging (MRI) occurred in 9 of 15 patients who could be evaluated (60%), including two complete responses. Peritumoral toxicity developed 1-4 weeks after treatment in three of three patients at 1.0 microg/ml, but in zero of nine patients treated at lower concentrations. No symptomatic systemic toxicity occurred. Regional perfusion with Tf-CRM107 produces tumor responses without systemic toxicity in patients with malignant brain tumors refractory to conventional therapy. Direct interstitial infusion can be used successfully to distribute a large protein in the tumor and infiltrated brain surrounding the tumor. PMID- 9396607 TI - Expression and function of CCR5 and CXCR4 on human Langerhans cells and macrophages: implications for HIV primary infection. AB - Transmission of HIV-1 is predominantly restricted to macrophage (Mphi)-tropic strains. Langerhans cells (LCs) in mucosal epithelium, as well as macrophages located in the submucosal tissues, may be initial targets for HIV-1. This study was designed to determine whether restricted transmission of HIV-1 correlates with expression and function of HIV-1 co-receptors on LCs and macrophages. Using polyclonal rabbit IgGs specific for the HIV co-receptors cytokines CXCR4 and CCR5, we found that freshly isolated epidermal LCs (resembling resident mucosal LCs) expressed CCR5, but not CXCR, on their surfaces. In concordance with surface expression, fresh LCs fused with Mphi-tropic but not with T-tropic HIV-1 envelopes. However, fresh LCs did contain intracellular CXCR4 protein that was transported to the surface during in vitro culture. Macrophages expressed high levels of both co-receptors on their surfaces, but only CCR5 was functional in a fusion assay. These data provide several possible explanations for the selective transmission of Mphi-tropic HIV variants and for the resistance to infection conferred by the CCR5 deletion. PMID- 9396608 TI - Complementary hydropathy identifies a cellular prion protein receptor. AB - Prions, the etiological agents for infectious degenerative encephalopathies, act by entering the cell and inducing conformational changes in PrPC (a normal cell membrane sialoglycoprotein), which result in cell death. A specific cell-surface receptor to mediate PrPC and prion endocytosis has been predicted. Complementary hydropathy let us generate a hypothetical peptide mimicking the receptor binding site. Antibodies raised against this peptide stain the surface of mouse neurons and recognize a 66-kDa membrane protein that binds PrPC both in vitro and in vivo. Furthermore, both the complementary prion peptide and antiserum against it inhibit the toxicity of a prion-derived peptide toward neuronal cells in culture. Such reagents might therefore have therapeutic applications. PMID- 9396609 TI - The human 37-kDa laminin receptor precursor interacts with the prion protein in eukaryotic cells. AB - Prions are thought to consist of infectious proteins that cause transmissible spongiform encephalopathies. According to overwhelming evidence, the pathogenic prion protein PrPSc converts its host encoded isoform PrPC into insoluble aggregates of PrPSc, concomitant with pathological modifications (for review, see refs. 1-3). Although the physiological role of PrPC is poorly understood, studies with PrP knockout mice demonstrated that PrPC is required for the development of prion diseases. Using the yeast two-hybrid technology in Saccharomyces cerevisiae, we identified the 37-kDa laminin receptor precursor (LRP) as interacting with the cellular prion protein PrPC. Mapping analysis of the LRP-PrP interaction site in S. cerevisiae revealed that PrP and laminin share the same binding domain (amino acids 161 to 180) on LRP. The LRP-PrP interaction was confirmed in vivo in insect (Sf9) and mammalian cells (COS-7). The LRP level was increased in scrapie-infected murine N2a cells and in brain and spleen of scrapie infected mice. In contrast, the LRP concentration was not significantly altered in these organs from mice infected with the bovine spongiform encephalopathic agent (BSE), which have a lower PrPSc accumulation. LRP levels, however, were dramatically increased in brain and pancreas, slightly increased in the spleen and not altered in the liver of crapie-infected hamsters. These data show that enhanced LRP concentrations are correlated with PrPSc accumulation in organs from mice and hamsters. The laminin receptor precursor, which is highly conserved among mammals and is located on the cell surface, may act as a receptor or co receptor for the prion protein on mammalian cells. PMID- 9396610 TI - Passive immunization with a human monoclonal antibody protects hu-PBL-SCID mice against challenge by primary isolates of HIV-1. AB - How well antibodies can protect against disease due to HIV-1 infection remains a pivotal but unresolved issue with important implications for vaccine design and the use of prophylactic antibody to prevent infection after accidental exposure to the virus and to interrupt transmission of virus from mother to child. Strong doubts about the possible utility of antibodies in vivo have been raised because of the relative resistance of primary viruses to antibody neutralization in vitro. Primary viruses are likely to be close to the viruses transmitted during natural infection in humans. Vaccine studies have been of little value in assessing antibody efficacy in vivo because none of the strategies described to date have elicited significant neutralizing antibody responses to primary viruses. Passive immunization studies are similarly hindered by the paucity of reagents able to neutralize primary viruses effectively and a single study has suggested some benefit. Here we describe experiments to explore the ability of passive antibody to protect against primary virus challenge in hu-PBL-SCID mice. In this model, severe combined immunodeficient (SCID) mice are populated with human peripheral blood mononuclear cells (PBMCs) and infected with HIV-1. We find that the potent neutralizing human monoclonal antibody IgG1b12 at high dose is able to completely protect even when given several hours after viral challenge. The results are encouraging for antibody-based postexposure prophylaxis and support the notion that antibody induction could contribute to an effective vaccine. PMID- 9396611 TI - Association of human herpes virus 6 (HHV-6) with multiple sclerosis: increased IgM response to HHV-6 early antigen and detection of serum HHV-6 DNA. AB - Viruses have long been suggested to be involved in the etiology of multiple sclerosis (MS). This suggestion is based on (1) epidemiological evidence of childhood exposure to infectious agents and increase in disease exacerbations with viral infection; (2) geographic association of disease susceptibility with evidence of MS clustering; (3) evidence that migration to and from high-risk areas influences the likelihood of developing MS; (4) abnormal immune responses to a variety of viruses; and (5) analogy with animal models and other human diseases in which viruses can cause diseases with long incubation periods, a relapsing-remitting course, and demyelination. Many of these studies involve the demonstration of increased antibody titers to a particular virus, whereas some describe isolation of virus from MS material. However, no virus to date has been definitively associated with this disease. Recently, human herpesvirus 6 (HHV-6), a newly described beta-herpes virus that shares homology with cytomegalovirus (CMV), has been reported to be present in active MS plaques. In order to extend these observations, we have demonstrated increased IgM serum antibody responses to HHV-6 early antigen (p41/38) in patients with relapsing-remitting MS (RRMS), compared with patients with chronic progressive MS (CPMS), patients with other neurologic disease (OND), patients with other autoimmune disease (OID), and normal controls. Given the ubiquitous nature of this virus and the challenging precedent of correlating antiviral antibodies with disease association, these antibody studies have been supported by the detection of HHV-6 DNA from samples of MS serum as a marker of active viral infection. PMID- 9396612 TI - Semaphorin III can repulse and inhibit adult sensory afferents in vivo. AB - During development, semaphorins (collapsin, fasciclin) mediate repulsive and inhibitory guidance of neurons. Semaphorin III, a secretable member of this family, is expressed by the ventral spinal cord at the time corresponding to projection of sensory afferents from the dorsal root ganglion (DRG) into the spinal cord. The inhibitory effect of E14 ventral cord is active only on nerve growth factor (NGF)-responsive sensory afferents (small-diameter A-delta and C fibers subserving sensations of temperature and pain). Similarly, COS cells secreting recombinant semaphorin III are able to selectively repel DRG afferents whose growth is stimulated by NGF and not NT-3. However, it is not known whether these molecules can exert a functional role in the fully developed adult peripheral nervous system. In this study, we demonstrated that gene gun transfection and production of semaphorin III in corneal epithelial cells in adult rabbits in vivo can cause repulsion of established A-delta and C fiber trigeminal sensory afferents. In addition, it is shown that, following epithelial wounding and denervation, semaphorin III is able to inhibit collateral nerve sprouts from innervating the reepithelialized tissue. These findings are significant in that they provide direct evidence that small-diameter adult sensory neurons retain the ability to respond to semaphorin III. In addition, the corneal gene gun technique may be generally used to study the in vivo effects of neural growth modulators by quantifying the amount of sensory nerve growth. PMID- 9396613 TI - Long-term gene therapy in the CNS: reversal of hypothalamic diabetes insipidus in the Brattleboro rat by using an adenovirus expressing arginine vasopressin. AB - The ability of adenovirus (Ad) to transfect most cell types efficiently has already resulted in human gene therapy trials involving the systemic administration of adenoviral constructs. However, because of the complexity of brain function and the difficulty in noninvasively monitoring alterations in neuronal gene expression, the potential of Ad gene therapy strategies for treating disorders of the CNS has been difficult to assess. In the present study, we have used an Ad encoding the arginine vasopressin cDNA (AdAVP) in an AVP deficient animal model of diabetes insipidus (the Brattleboro rat), which allowed us to monitor chronically the success of the gene therapy treatment by noninvasive assays. Injection of AdAVP into the supraoptic nuclei (SON) of the hypothalamus resulted in expression of AVP in magnocellular neurons. This was accompanied by reduced daily water intake and urine volume, as well as increased urine osmolality lasting 4 months. These data show that a single gene defect leading to a neurological disorder can be corrected with an adenovirus-based strategy. This study highlights the potential of using Ad gene therapy for the long-term treatment of disorders of the CNS. PMID- 9396615 TI - Mass spectrometry from miniaturized arrays for full comparative DNA analysis. PMID- 9396614 TI - PECAM-1/CD31, an endothelial receptor for binding Plasmodium falciparum-infected erythrocytes. AB - Excessive binding of Plasmodium falciparum-infected red blood cells (pRBCs) to the vascular endothelium (cytoadherence) and to uninfected erythrocytes (rosetting) may lead to occlusion of the microvasculature and thereby contribute directly to the acute pathology of severe human malaria. A number of endothelial receptors have been identified as targets for the pRBCs, including CD36, intercellular adhesion molecule-1 (ICAM-1) and chondroitin-4-sulfate (CSA). In vitro, CD36 is the most frequent target of strains from patients with mild as well as severe P. falciparum malaria, but is expressed at low levels on the cerebral microvasculature and therefore seems unlikely to be involved in the evolution of cerebral disease. Strains of P. falciparum that form rosettes are associated both with the occurrence of cerebral malaria and severe anemia. Here we report that malaria-infected RBCs adhere to platelet/endothelial cell adhesion molecule-1 (PECAM-1/CD31) on the vascular endothelium. pRBCs bind to endothelial cells, to PECAM-1/CD31 transfected cells, and directly to recombinant PECAM 1/CD31 absorbed onto plastic. Soluble PECAM-1/CD31 and monoclonal antibodies specific for the amino-terminal segment of PECAM-1/CD31 (domains 1-4) blocked the binding. Interferon-gamma (IFN-gamma)-essential for the development of cerebral malaria in the mouse-was found to augment adhesion of human pRBCs to PECAM-1/CD31 on endothelial cell monolayers. Our results suggest that PECAM-1/CD31 is a virulence-associated endothelial receptor of P. falciparum-infected RBCs. PMID- 9396616 TI - Cancer vaccines. AB - A better understanding of immune recognition of cells has led to identification of potential new targets on tumor cells. Noticeable successes in melanoma have been immunization with the GM2 ganglioside vaccine, and the identification of novel antigens such as MAGE, BAGE and GAGE recognized by T cells cloned from cancer patients with regressing disease. However, the unexpected finding that other antigens recognized by these T cells were overexpressed normal differentiation antigens such as tyrosinase. Pmel 17 and Melan A have led to vaccines developed against differentiation antigens expressed in other solid tumors. Monoclonal antibody, anti-idiotype and antigen based vaccines for colorectal target antigens 17-1A, CEA and 791Tgp72 are all in clinical development. Similarly HER2/neu and mucin overexpression in breast cancer represent promising targets. Mutations in tumor oncogenes or suppressor genes which lead to malignant transformation can also present tumor-specific antigens. The most effective vaccines against infectious disease are live viruses. The development of DNA vaccines which act like viruses in entering cells and show continuous production of antigens offers great potential for the future. PMID- 9396617 TI - Role of vindesine as neoadjuvant chemotherapy for non-small cell lung and head and neck cancers. AB - Although surgery is the only therapeutic intervention with potential for cure of non-small cell lung cancer (NSCLC) and head and neck cancer, most patients present with tumors that are too far advanced for resection. For this reason, neoadjuvant chemotherapy is being increasingly used to down-stage primary tumor burden before surgery. The procedure offers the possibility of enhancing resectability and thereby improves the chances of achieving complete eradication. In NSCLC, the most successful results have been obtained in patients with disease that is localized to the thorax and in those achieving complete responses to chemotherapy. A number of different combination regimens have been studied with cisplatin as the key component. Response rates of up to 88% have been reported together with complete sections in up to 74% of patients. The impact on survival is still to be determined. Although there is ample data showing that combination regimens containing vindesine have a favorable effect on survival in patients with inoperable NSCLC, relatively few of the studies have evaluated these regimens for use as neoadjuvant therapy. New studies should focus on increasing the complete response rate above the 20% level that is currently attainable. The results using neoadjuvant chemotherapy in head and neck cancer are disappointing with no survival benefit demonstrated. However, neoadjuvant chemotherapy may help preserve organ function and thus improve quality of life of patients. PMID- 9396618 TI - Eriochrome Black T, structurally related to suramin, inhibits angiogenesis and tumor growth in vivo. AB - The polyanionic species suramin is a potential anti-cancer agent of narrow therapeutic index. Among other pharmacological characteristics, suramin is an inhibitor of angiogenesis. We have targeted its angiostatic properties as part of a program to discover less toxic analogs. From screening a series of commercially available compounds, structurally related to suramin and containing a sulfonic acid substituted naphthylamine moiety, we discovered a new lead, Eriochrome Black T (EBT). EBT is a novel inhibitor of angiogenesis, more potent and less toxic than suramin in the chick chorioallantoic membrane assay. EBT was more active than suramin in inhibiting endothelial cell proliferation in primary culture and in inhibiting proliferation of three tumor cell lines, A431, L1210 and M5076 (IC50 10-100 microM). Cell cycle studies on the A431 line showed that both EBT and suramin caused an accumulation of cells in the S phase, EBT being 10-fold more potent. We suggest that this cell cycle perturbation is linked to inhibition of topoisomerase II catalytic activity. EBT was found to be a moderate but significant inhibitor of matrix metalloproteinases (10 microM range), more efficient than suramin. In a s.c. M5076 sarcoma model in mice, EBT had similar efficacy to suramin both by the i.p. or s.c. route and was moreover better tolerated. Combined pharmacological results show that EBT compared favorably with suramin in all assays, and that in ovo and in vivo, EBT is an analog of suramin with diminished toxicity. PMID- 9396619 TI - The ex vivo chemosensitivity profile of choroidal melanoma. AB - Uveal melanoma has a high mortality rate and responds poorly to existing chemotherapy. We have therefore examined the sensitivity of uveal melanoma to cytotoxic agents using an ex vivo chemosensitivity assay to determine whether some agents which have not been used for this tumor might have activity. An ATP based tumor chemosensitivity assay (ATP-TCA) was used to determine the effect of nine cytotoxic drugs at multiple dilutions in 28 primary uveal melanoma specimens. Evaluable assay results from up to 16 tumors with each drug showed variable sensitivity to alkylating agents (three of nine with mitomycin C, one of 15 with cisplatin and seven of 15 with treosulfan), cytosine arabinoside (seven of 16), paclitaxel (one of five) and doxorubicin (two of 16). No tumors were sensitive to temozolomide, or 5-fluorouracil, and only one of 14 to vincristine. The combination of treosulfan with cytosine arabinoside resulted in enhanced tumor cell inhibition in three of five tumors tested. Combinations containing paclitaxel combined with doxorubicin or cisplatin showed some activity, but none achieved 100% inhibition and the results were similar to those obtained with paclitaxel alone. Uveal melanoma shows considerable heterogeneity of sensitivity to cytotoxic drugs, with considerable resistance to most agents, matching clinical experience. The results suggest that cytosine arabinoside or gemcitabine plus treosulfan may be an active combination. Clinical trials will commence shortly. The use of the ATP-TCA provides a method of testing multiple agents and combinations in a way which would be otherwise impossible in rare tumors such as uveal melanoma. PMID- 9396620 TI - Schedule-dependent paclitaxel tolerance/activity: data from a 7 day infusion phase I study with pharmacokinetics in paclitaxel refractory ovarian cancer. AB - Our objective was to determine the maximum tolerated dose (MTD) of paclitaxel when given as a 7 day continuous i.v. infusion, repeated every 3 weeks, and to evaluate the toxicity and the efficacy of such a schedule of administration as a salvage treatment in ovarian cancer patients pretreated and refractory to 3 or 24 h paclitaxel. Thirteen women were enrolled in this phase I trial. Four dose levels ranging from 105 to 157.5 mg/m2/cycle were explored. Two of four patients experienced dose-limiting febrile neutropenia at the dose of 157.5 mg/m2. No objective response was observed, although three patients experienced disease stabilization (five to six cycles), with regression of disease symptoms, two of them having sustained 50% or greater decrease in CA 125. We conclude that the MTD in this population was paclitaxel 140 mg/m2/7 days. Schedule-dependent mechanisms of resistance to paclitaxel could not be demonstrated in this clinical setting of heavily pretreated ovarian cancer patients. PMID- 9396621 TI - Evaluation of methotrexate sensitivity in human leukemia cell lines by an adenosine triphosphate bioluminescence assay. AB - To verify a recently developed in vitro tumor chemosensitivity assay (TCA) based on adenosine triphosphate (ATP) measurement for detection of methotrexate (MTX) sensitivity or resistance in human leukemias and solid tumors in which the antifol is indicated, we investigated the ability of the assay to discriminate between sensitivity and resistance to this antifolate in human leukemia cell lines sensitive (parental CCRF-CEM/S) and resistant (CCRF-CEM/E, CCRF-CEM/T and CCRF-CEM/P sublines) to MTX by virtue of known biochemical mechanisms. Correlation experiments with a standard cell growth inhibition assay and a radiometric method for measurement of thymidylate (dTMP) synthesis ([5-3H]-2' deoxyuridine tritium release assay) were performed. No significant differences were observed in the IC50 values for the four cell lines tested as determined by cell growth evaluation (cell number counts) and the ATP-TCA after a 72 h MTX exposure. After short-term (4 h) high-dose MTX exposure, no significant correlation between ATP-TCA and the classic [5-3H]-2'-deoxyuridine tritium release assay was observed in both CCRF-CEM/S and CCRF-CEM/P cells. CCRF-CEM/T and CCRF-CEM/E displayed, instead, complete resistance with both methods. When using conditions proposed for clinical application (long-term exposure, i.e. 144 h) the ATP-TCA permitted the identification of cell lines highly resistant to MTX (CCRF-CEM/F and CCRF-CEM/E), while intermediate MTX resistance due to altered polyglutamylation was not detectable. Detection of this kind of resistance was obtained, as expected, using a short-term exposure (4 h) to MTX followed by a long-term efflux (72 h) in drug-free medium. On the basis of these results, ATP TCA appears to be a suitable method for the evaluation of cytotoxicity induced by MTX. PMID- 9396622 TI - Potentiation of the cytotoxicity of carboquone by flavone acetic acid combined with hyperthermia. AB - Flavone acetic acid (FAA) has shown the effectiveness of vasoactive drugs in the selective reduction of tumor blood flow. A FAA-mediated decrease in tumor blood flow may produce sufficient hypoxic conditions within the tumor. Carboquone (CQ), a naturally occurring prototype bioreductive alkylating agent like mitomycin C (MMC), has been shown to be selectively more cytotoxic toward hypoxic tumor cells. We have reported enhancement of the combined antitumor effects of MMC plus FAA and hyperthermia (HT). In this study, we examined the combined effects of FAA, CQ and HT. In vitro, although HT (43 degrees C, 60 min) reduced the colonogenicity to 0.58 in CQ (0.01 microg/ml) alone, the combined cytotoxicity of CQ and HT was not enhanced with exposure to FAA at a concentration of 100 microg/ml. In vivo, the tumor growth time, calculated as the time required to reach twice the initial tumor volume, for CQ (2 mg/kg) alone, FAA (150 mg/kg) alone, CQ+FAA, CQ+HT (43 degrees C, 15 min), FAA+HT and FAA+CQ+HT was 6.1, 5.1, 7.1, 8.0, 7.6 and 13.4 days, respectively. A significant enhancement of antitumor effects by trimodality therapy with CQ, FAA and HT was observed, when compared to the treatment with CQ and FAA (p < 0.05). The possible mechanisms of an increased antitumor response achieved with the combination of CQ, FAA and HT may be explained in the following way: the FAA-mediated decrease in tumor blood flow produced sufficient hypoxic conditions within the tumor, and these resulted in a significant increase of the antitumor effects of CQ and HT. PMID- 9396623 TI - Activation of murine peritoneal macrophages after cisplatin and taxol combination. AB - Cisplatin and paclitaxel are potent antineoplastic agents. Their distinctly different mechanisms of action have prompted laboratory and clinical research into their use in combination therapies. Murine peritoneal macrophages treated with cisplatin and paclitaxel in combination elicit an increase in their number of lysosomes. Drug-treated macrophages, when co-incubated with sarcoma 180 cells, establish cytoplasmic contact and transfer lysosomes into tumor cells causing tumor cell lysis. In addition, analysis of tissue culture supernatants show increased levels of interleukin-1alpha and tumor necrosis factor-alpha. Our study shows that cisplatin and paclitaxel in combination enhance elements of the immune system with greater efficacy and potency than when used alone. PMID- 9396624 TI - 5-Fluorouracil + cisplatin + mitomycin C is a relatively most effective combination against xenograft lines of human colorectal cancer. AB - 5-Fluorouracil (5-FU) has been an accepted effective against colorectal cancer, but combination regimens resulted in a lesser effect than 5-FU alone. The present study was designed to evaluate the efficiency of various combination chemotherapies, including 5-FU, on human colorectal cancer xenograft lines (CC-KK and RC-TK), which had been passed by transplantation in nude mice. Eight anticancer agents [5-FU, mitomycin C (MMC), adriamycin (ADR), 4'-epirubicin (EPIR), carboquone (CQ), cisplatin, carboplatin and etoposide (VP-16)] and their combinations were evaluated at 2-4 times the clinical dose. When the agents were administered singly, 5-FU, EPIR and CQ were effective against CC-KK, and 5-FU, MMC, EPIR, ADR, CQ and carboplatin were effective against RC-TK. Of the two-agent combinations including 5-FU, cisplatin + 5-FU was the most effective against both CC-KK and RC-TK. However, of the three-agent combinations, only cisplatin + 5-FU + MMC was more effective against both lines than cisplatin + 5-FU. These results suggest that cisplatin + 5-FU + MMC may be the most useful regimen against colorectal cancers in clinical application. PMID- 9396625 TI - Cisplatin and interferon-gamma treated murine macrophages induce apoptosis in tumor cell lines. AB - Macrophages, treated in vitro with a combination of cisplatin and interferon (IFN)-gamma, have been shown to develop enhanced tumoricidal activity against a number of tumor cell types, through mechanisms which remain largely unknown. In the present study, we have investigated the mechanism involved in the tumor cell cytotoxicity mediated by cisplatin and lFN-gamma treated macrophages, and the effector molecules involved therein. Peritoneal macrophages treated with cisplatin and IFN-gamma, when co-cultured with different tumor cell types, caused tumor cell death by induction of apoptosis. Evidence for this was provided by percent specific DNA fragmentation assay, by specific pattern of internucleosomal DNA fragmentation detected by agarose gel electrophoresis and by microscopic examination of the cells, which revealed nuclear alterations, characteristic of apoptosis. The time kinetics studies of DNA fragmentation, loss in cell viability and apoptotic cell population showed linearity with time; most of the cells that underwent apoptosis were found to be viable even after 24 h co-culture. Macrophages induced apoptosis in tumor targets even in the absence of cell-to cell contact, i.e. via diffusible effector molecules. In P815 cells, NO produced by cisplatin and IFN-gamma treated macrophages was found to induce apoptosis as addition of N(G)-monomethyl L-arginine (L-NMMA), a specific inhibitor of NO synthase to the co-culture, prevented apoptosis in P815 cells. Further, direct treatment of P815 cells with the NO donor, sodium nitroprusside (SNP), resulted in apoptosis. In L929 cells, the effector molecule was found to be tumor necrosis factor (TNF)-alpha as apoptosis was blocked by the addition of anti-TNF-alpha antibodies to the co-culture but the addition of L-NMMA or SNP had no effect. The study thus shows that cisplatin and IFN-gamma treated macrophages can kill tumor cells by extracellular release of effector molecules which act by inducing apoptosis in a target cell-specific manner. PMID- 9396626 TI - Molecular characterization of a cDNA encoding a novel small GTP-binding protein from Arabidopsis thaliana. AB - A full-length cDNA clone encoding a novel Rab protein AtRab alpha of the monomeric small GTP-binding protein family has been isolated from Arabidopsis thaliana. AtRab alpha has 210 amino acids with a calculated molecular mass of 23.3 kDa. The highest homology was found to Rab1x and Rab1y from Lotus japonicus. Southern blot analysis of genomic DNA indicated that AtRab alpha was encoded by a single copy gene. Northern blot analysis showed that expression of the AtRab alpha mRNA was rich in stems and roots, but poor in leaves and flowers, which is different from the expression pattern of other Arabidopsis Rab genes. PMID- 9396627 TI - One allele of the major histone gene cluster is enough for cell proliferation of the DT40 chicken B cell line. AB - Thirty-nine of the 44 chicken histone genes are located in a major histone gene cluster of 110 kb, the others residing in four separate regions. We generated a heterozygous chicken DT40 mutant, 1/2 delta110 kb, devoid of one allele of the cluster, using gene targeting techniques. Analyses of the mutant revealed that the growth rate of DT40 cells was unchanged even in the absence of one allele of the cluster. Moreover, analyses involving a RNase protection assay, SDS-PAGE or Triton-acid-urea-PAGE revealed not only that in the 1/2 delta110 kb mutant the steady-state levels of total mRNAs of gene families H1, H2A, H2B, H3 and H4 remained constant, but also that the amounts of histones H1, H2A, H2B, H3 and H4 were not changed. A comparison by 2D-PAGE revealed no changes in total cellular protein patterns of the mutant. These observations demonstrate that all the histone gene families have the inherent ability to compensate for the disruption of one allele of the gene cluster, with no influence on cell functions. PMID- 9396628 TI - Expression of Tetrahymena histone H4 in yeast. AB - Histone H4 is one of the most conserved proteins known. The very low rate of nonsynonymous substitution in H4 suggests that it fulfills an essential function in virtually all eukaryotes. While the majority of histone H4 sequences differ only slightly from the general consensus H4 sequence, yeast and Tetrahymena sequences diverge substantially from both the consensus and from each other. This study demonstrates that despite this divergence, when Saccharomyces cerevisiae cells are forced to use the Tetrahymena thermophila histone H4 protein, they are viable although they have a reduced growth rate, are temperature-sensitive relative to wild-type, have a lengthened G2 phase, and show a dramatic repression of mating. An amino acid replacement at position 33 in the protein improves the growth rate of these cells growing at temperatures above 28 degrees C. This replacement changes a proline to a serine and is a further divergence from both the Tetrahymena thermophila and Saccharomyces cerevisiae histone H4 sequences. Thus, the replacement and expression of a non wild-type histone H4 in yeast offers measurable effects on cell growth, identifying amino acids required for optimal yeast functioning. PMID- 9396629 TI - Head-to-head linkage of carp alpha- and beta-globin genes. AB - The alpha- and beta-globin gene variants are believed to have diverged from a single ancestral globin gene, and the divergence was primed by the duplication of the ancestral globin gene. To understand the process of gene duplication, we investigated the alpha- and beta-globin gene arrangement of a bony fish (carp). From a Southern analysis of seven previously prepared lambda phage clones (lambdaCG1-7) using radio-labelled alpha- or beta-globin gene probes, it was found that the clones included both the alpha- and beta-globin genes, and that they were located within a distance of 1 kb. Additionally, the linkage of two alpha-globin genes and two beta-globin genes in the clone lambdaCG1, 5 and 7 (e.g., alpha-beta-alpha-beta in lambdaCG5) revealed an arrangement that is different from the arrangement in higher vertebrates in which the alpha-globin and beta-globin genes generally occur at different loci. The distances between the detached alpha- to beta-globin genes were approximately 5 to 10 kb. DNA sequencing of the adjacently linked alpha- and beta-globin genes in lambdaCG3 showed that they were arranged in a head-to-head orientation. PCR amplification using primers for the internal region between the carp alpha- and beta-globin genes gave approximately 0.9-kb products from each of the clones lambdaCG1, 3, 4, 5, 6, and 7, and from the chromosomal DNA of German mirror carp, Saku carp, Suwa carp, and Yamato carp. This demonstrates the alternative arrangement of carp alpha- and beta-genes in the globin gene locus (i.e., 3'alpha5'-5'beta3' 3'alpha5'-5'beta3' in lambdaCG5), and the widespread distribution of head-to-head linked alpha- and beta-globin genes in carp. Based on the above results, we hypothesize that the duplication of the ancestral globin gene (prior to the divergence of the a and beta forms) occurred in a head-to-head direction. PMID- 9396630 TI - The human zinc finger protein EGR-4 acts as autoregulatory transcriptional repressor. AB - The human EGR-4 (AT133) gene represents one member of a family of four related zinc finger proteins, that are simultaneously and coordinately induced in resting cells upon growth stimulation. In order to characterise the function of the EGR-4 zinc finger protein, we have expressed the protein in the eukaryotic baculovirus system. The recombinant EGR-4 protein has a molecular mass of 78 kDa, as demonstrated by SDS-PAGE and Western blotting. DNA binding studies revealed that the EGR-4 protein binds to the EGR consensus motif GCGTGGGCG, but not to the G rich regulatory ZIP-element of the human IL-2 gene, that is a binding site for EGR-1. EGR-4 functions as transcriptional repressor. Overexpression of EGR-4 mediates repression of a minimal c-fos promoter through a threefold EGR consensus site. Furthermore the EGR-4 protein displays autoregulatory activities. This protein downregulates expression of its own gene promoter in a dose dependent manner. A G-rich region in the EGR-4 promoter, located at position -106 to -82, could be identified as binding site for the recombinant EGR-4 protein. A comparison of the two related zinc finger proteins EGR-4 and EGR-1 revealed for each protein distinct and specific DNA binding- and transcriptional regulatory activities. PMID- 9396631 TI - Fluorescence and NMR studies of intramolecular stacking of mRNA cap-analogues. AB - Intramolecular stacking of a series of new synthesized dinucleotide mRNA cap analogues has been investigated in aqueous buffers by means of fluorescence and 1H-NMR at various pH and temperatures, and compared with that for 7 methylguanosine(5')ppp(5')guanosine (m7GpppG), as well as its hypermethylated derivative m(3)2,2,7GpppG. Thermodynamic parameters for intramolecular self association stabilized by stacking were established by temperature-dependent fluorescence quenching, taking into account collisional deactivation of the excited states. Relative orientations of the stacked bases in the cap analogues were determined with the aid of a program GEOSHIFT (Stolarski et al., Biochim. Biophys. Acta (1996) 1293, 97), based on ring-current anisotropy. 1D-soft-TOCSY experiments were applied to extract the exact values of vicinal coupling constants, and hence to resolve solution conformation of the cap molecules. Stacking interaction has been discussed in detail in terms of the cap structural features, e.g., types of bases and length of the 5',5'-phosphate bridges, and regarding the interactions stabilizing intramolecular stacking. PMID- 9396632 TI - Chinese hamster ovary cells lacking GM1 and GD1a synthesize gangliosides upon transfection with human GM2 synthase. AB - GM3-positive Chinese hamster ovary cells (CHO-K1 cells) lack the ability to synthesize GM2 and the complex gangliosides GM1 and GD1a from [3H]Gal added to the culture medium. However, they acquire the ability to synthesize GM2 and to synthesize and immunoexpress complex gangliosides upon transient transfection with a cDNA encoding the human GM3:N-acetylgalactosaminyl transferase (GM2 synthase). The activities of endogenous GM1- and GD1a-synthases in the parental cell line and in cells transfected with the plasmid with or without the GM2 synthase cDNA were essentially identical and comparable in terms of specific activity with the endogenous GM3 synthase. Results indicate that glycosyltransferases acting on GM2 to produce GM1 and GD1a are constitutively present in CHO-K1 cells, and that the expression of their activities depend on the supply of the acceptor GM2. In addition, these results lend support to the notion that GM2 synthase is a key regulatory enzyme influencing the balance between simple and complex gangliosides. PMID- 9396633 TI - Identification of soluble type of membrane-type matrix metalloproteinase-3 formed by alternatively spliced mRNA. AB - Homology screening for human membrane-type MMP (MT-MMP) was carried out, and cDNA encoding a soluble type of MT3-MMP (SM3), which is considered to be an alternatively spliced variant of MT3-MMP, was obtained. SM3 had a novel sequence consisting of 50 amino acids after Lys407 instead of amino acids containing the transmembrane domain of MT3-MMP. When SM3 tagged with a FLAG epitope (SM3-flag) was expressed in COS-7 cells, SM3-flag was present in the conditioned medium in its activated form. The enzymatic activity of SM3 was studied using a recombinant enzyme expressed in E. coli (SM3-e). The fluorogenic peptide substrate hydrolyzing activity of SM3-e was inhibited by EDTA and by the tissue inhibitor of metalloproteinase-2 (TIMP-2), whereas TIMP-1 had only relatively weak inhibitory ability. SM3-e was able to activate proMMP-2, and this activity was also inhibited by TIMP-2 but not by TIMP-1. SM3-e was able to cleave type III collagen, and also digested fibronectin. In view of the homology of the primary structures, MT3-MMP was considered to have the same catalytic activity as SM3. The results of studies of SM3's activity on extracellular matrix (ECM) protein suggests that MT3-MMP plays a role in ECM turnover not only by activating proMMP 2 but also by acting directly on ECM macromolecules. PMID- 9396635 TI - Triggering and the pathophysiology of acute coronary syndromes. AB - Insight into the pathophysiology of acute coronary syndromes can be gained by studying the role of triggering activities, such as heavy physical exertion and episodes of anger, in promoting plaque rupture and thrombosis, and the determinants of plaque vulnerability to disruption, which include lipid-rich plaque, a thin fibrous cap, and increased macrophage activity. This article focuses on the contribution of triggering activities and the potential mechanisms by which they may lead to acute myocardial infarction and sudden cardiac death. PMID- 9396634 TI - Transfection of TTF-1 gene induces thyroglobulin gene expression in undifferentiated FRT cells. AB - The thyroglobulin gene, the substrate for thyroid hormone biosynthesis, is not expressed in the FRT cell line, which, even though it manifests the polarised epithelial phenotype, does not express any of the thyroid functional properties. Two transcription factors, TTF-1 and Pax-8, have been implicated in thyroid specific expression of the thyroglobulin gene. FRT cells contain Pax-8 but they lack TTF-1. In this paper, we show that transfection of TTF-1 expression vectors in FRT cells results in activation of thyroglobulin gene expression. If the expression vector encoded for TTF-1-ER, a fusion gene coding for the entire TTF-1 protein fused to the hormone-binding domain of the steroid receptor, under the control of the RSV promoter, thyroglobulin gene expression was controlled by estrogen. These data provide a direct demonstration that TTF-1 activates the chromosomal thyroglobulin promoter. Since transfection of TTF-1 expression vectors in non-thyroid cell types did not result in thyroglobulin gene expression, it is suggested that Pax-8, in addition, perhaps, to a specific cellular environment, might be required for thyroid specific expression of the thyroglobulin gene. PMID- 9396636 TI - Antiplatelet therapy: do the new platelet inhibitors add significantly to the clinical benefits of aspirin? AB - The inhibitors of the platelet membrane glycoprotein IIb/IIIa show considerable promise as antiplatelet agents. These drugs are easily titrated when administered intravenously and are associated with less frequent and serious bleeding than initially feared. They add significant benefit to that attributable to aspirin in preventing the complications associated with coronary angioplasty. Pilot studies have suggested that benefits could also be realized in acute myocardial infarction and unstable angina. The most effective means of administering these agents, their relative efficacy, and the consequences of long-term modulation of the glycoprotein IIb/IIIa receptor by oral agents are challenging areas for clinical investigation. PMID- 9396637 TI - Successors to heparin: new antithrombotic agents. AB - New advances in antithrombotic therapy include low-molecular weight heparins (LMWHs), heparinoids, and direct thrombin inhibitors. LMWHs and heparinoids have improved pharmacologic and pharmacokinetic properties compared with standard, unfractionated heparin. LMWHs are effective in preventing venous thromboembolism in general surgical patients, orthopedic patients, and general medical patients. At equipotent antithrombotic doses, LMWHs produce less bleeding than does unfractionated heparin. When given in fixed doses subcutaneously in the treatment of venous thromboembolic disease, LMWHs have been shown to be as effective as or more effective than and safer than standard heparin given intravenously with regular monitoring. Recent studies have demonstrated that LMWHs are effective in reducing the risk of death and myocardial infarction in patients with unstable angina; they are as effective as intravenous heparin when given subcutaneously without monitoring. Preliminary data indicate that LMWHs also may be effective in improving outcomes in patients with ischemic stroke. The pharmacokinetic characteristics of LMWHs permit their use in a fixed dose administered subcutaneously without monitoring, resulting in greater clinical utility than standard heparin. The direct thrombin inhibitors have a narrow safety window, and in doses that do not produce excessive bleeding have not been shown to have greater efficacy than that of unfractionated heparin. PMID- 9396638 TI - The evolution of antithrombotic therapy in coronary stenting. AB - A rare but important side effect associated with the use of coronary stents is the development of subacute thrombotic occlusion within the first week after implantation. Recent studies have indicated that the incidence of subacute thrombosis can be reduced with a combination regimen consisting of aspirin and ticlopidine instead of the standard regimen of aspirin and warfarin. The evolution in our understanding of the factors causing subacute stent thrombosis, coupled with the development of new implantation procedures, has permitted the development of a new treatment strategy with less aggressive anticoagulation therapy, fewer side effects, and, it is hoped, lower costs. PMID- 9396639 TI - Rationale for low-molecular weight heparin in coronary stenting. AB - Stents have been as revolutionary for the practice of coronary revascularization in recent years as was the coronary angioplasty balloon 15 years ago, but they have also been associated with a high rate of stent thrombosis. The Enoxaparin and Ticlopidine After Elective Stenting (ENTICES) trial is designed to determine the impact of a reduced anticoagulation regimen on clinical outcomes after stent deployment. Patients are randomly assigned 2:1 to enoxaparin-ticlopidine-aspirin versus the conventional warfarin regimen, and surrogate markers of platelet activation and thrombin activity are measured after 3 days. Three factors underpin ENTICES: (1) a desire to eliminate stent thrombosis, (2) a desire to reduce length of stay after stent placement by avoiding the prolonged hospitalization required with the five-drug regimen of heparin, aspirin, dipyridamole, dextran, and warfarin, and (3) a desire to reduce the bleeding complications associated with the intense anticoagulation typically used in patients receiving stents. Patients are enrolled at seven sites in the United States and include patients with recent infarctions, restenotic lesions, and lesions as large as 30 mm in length. Other trials have also addressed issues concerning anticoagulation in patients undergoing stenting. The Stent Antithrombotic Regimen Study (STARS) trial compared aspirin, aspirin plus ticlopidine, and aspirin plus warfarin in 1650 patients receiving stents. The Aspirin/Ticlopidine vs Low-Molecular Weight Heparin/Aspirin/Ticlopidine High-Risk Stent Trial (ATLAST) is comparing aspirin plus ticlopidine with enoxaparin, aspirin, and ticlopidine in a group of patients at high risk undergoing stenting. The Intracoronary Stenting and Antithrombotic Regimen (ISAR) trial, a trial of ticlopidine, aspirin, and 12 hours of postprocedural heparin versus phenocoumaron on and aspirin after stenting in 517 patients, found a significantly lower incidence of the combined end point of death, myocardial infarction, bypass surgery, or repeated percutaneous transluminal angioplasty in the patients who received antiplatelet therapy, but the patients enrolled were not representative of the usual population undergoing stenting. New trials of stents and their sequelae should include low-molecular weight heparins and should gather cost and outcome data to satisfy capitated systems and managed care. Innovative stent designs may also permit changes in antithrombotic regimens. PMID- 9396640 TI - Issues of cost-effectiveness in the use of antithrombotic therapy for ischemic heart disease. PMID- 9396641 TI - The biology of the immune system. AB - Intact immunity is fundamental for survival. The human immune system has evolved with the sophisticated biologic capacity to distinguish self from nonself and for memory through the process of clonal expansion. The ability to distinguish even subtle differences from self, and among myriad antigens, is possible by the rearrangement of genes that encode immunoglobulins and T-cell receptors, as well as by the requirement for T cells to recognize antigens in the context of presentation by HLA molecules encoded within the major histocompatibility complex. Modulation of immune function initiated by antigenic stimulation and cell-cell interactions is facilitated by a plethora of soluble mediators such as cytokines. This overview of the biology of the immune system provides a framework for understanding physiologic immune responses and how lacunar defects within the immune system explain the pathogenesis of immunologic disorders. Through such understanding, potential targets can be identified for therapeutic modulation of the immune system. PMID- 9396642 TI - The cells of the allergic response: mast cells, basophils, and eosinophils. AB - Mast cells, basophils, and eosinophils have long been regarded as important effector cells in allergic disorders. Indeed, it is thought that the cells' cytoplasmic granule-associated or lipid mediators contribute to many of the signs and symptoms that are characteristic of these diseases. Mast cells, basophils, and eosinophils also probably contribute to protective host responses, especially to parasites. In addition, recent evidence shows that mast cells, basophils, and eosinophils can secrete a wide spectrum of cytokines and, in some cases, express functions that may permit them to regulate the development or perpetuation of allergic responses. Thus, mast cells, basophils, and eosinophils may express immunoregulatory activities, as well as serve as effector cells. PMID- 9396643 TI - Introduction to diagnostic laboratory immunology. AB - Assays performed in the diagnostic immunology laboratory support the diagnosis and management of a wide spectrum of clinical conditions. This chapter reviews immunologic principles as they apply to diagnostic laboratory assays. Most of the determinations are based on well-established principles of antigen-antibody reactions. Some of the specific areas discussed include flow cytometric analyses, critical in the care of patients with hematologic malignancies, with the acquired immunodeficiency syndrome, or undergoing transplantation, and protein electrophoretic assays to identify the presence of monoclonal gammopathies. We also discuss the use of molecular techniques in the diagnosis of hematologic malignancies and primary immunodeficiencies, characterization of the major histocompatibility complex, and enumeration of viral burden. PMID- 9396644 TI - Primary immunodeficiency diseases. AB - Primary immunodeficiencies are rare, but important for 3 reasons. First, a high index of suspicion and prompt diagnosis can lead to lifesaving treatment or significant improvement in quality of life. Second, appreciation of the genetic nature of a host defense defect makes possible family counseling and carrier and prenatal diagnosis. Finally, the large and growing list of human genetic defects in immune pathways provides an important tool for understanding human immunoregulation. Many inherited immunodeficiency diseases have had their genetic cause proven with the discovery of their disease genes within the past 5 years. These diseases provide a framework into which additional diseases and disease gene discoveries can be added as the rapid progress in molecular immunology and genetics continues. PMID- 9396645 TI - Rhinitis and inhalant allergens. AB - Allergic rhinitis affects about 20% of the US population. The diagnosis is based on patterns of symptoms, physical examination, and assessment of IgE antibodies by skin or in vitro testing. The most common offending allergens are pollens of grasses, trees, and weeds; fungi; animal allergens; and dust mites. In an individual with nasal allergy, exposure leads to rapid release of mast cell derived mediators. This immediate response is followed by a cell-dominated response, including eosinophils and lymphocytes. Cytokines from T(H)2 lymphocytes, such as interleukin 4 and interleukin 5, orchestrate allergic inflammation. Resulting tissue changes produce symptoms of the disease and augment responses on subsequent exposure to allergens and irritants. Strategies for avoiding offending agents are important in management. In intermittent disease, antihistamines and/or decongestants are first prescribed. More continuous symptoms may mandate intranasal steroids. Immunotherapy is often helpful for patients who respond poorly to pharmacotherapy and avoidance. PMID- 9396646 TI - Nasal polyps and sinusitis. AB - Despite the prevalence and long history of nasal polyps, many questions still exist with respect to incidence and pathogenesis. Although allergy has been commonly thought to be a major cause, much compelling evidence argues against this. Medical therapy consists of a short course of systemic steroids followed by intranasal steroids. Sinusitis is the most commonly reported chronic disease in the United States. Decrease in ostial size, retention of secretions, and decrease in mucociliary action all contribute to the pathogenesis of sinusitis. The clinical presentation of chronic sinusitis is generally subtle and the clinical index of suspicion must be high. Limited coronal computed tomography is regarded as the most definitive and cost-effective imaging technique for the diagnosis of sinusitis. Appropriate antibiotics must be administered for a sufficient period. In medically resistant sinusitis, functional endoscopic sinus surgery has emerged as the procedure of choice. PMID- 9396647 TI - Asthma. AB - For unknown reasons, the morbidity and mortality from asthma are increasing in many parts of the world, making it a global health concern. The heterogeneous nature of the clinical manifestations and therapeutic responses of asthma in both adult and pediatric patients indicate that it may be more of a syndrome rather than a specific disease entity. Numerous factors, including viral infections, allergen and irritant exposure, and exercise, among others, complicate both the short- and long-term management of asthma. Therapeutic intervention has focused on the appreciation that airway obstruction in asthma consists of bronchial smooth muscle spasm and variable degrees of airway inflammation characterized by edema, mucous secretion, and the influx of a variety of inflammatory cells. Choosing appropriate medications depends on the disease severity (intermittent, mild persistent, moderate persistent, severe persistent), patterns of disease activity (exacerbations related to viruses, allergens, exercise, etc), and the age at onset (infancy, childhood, adulthood). PMID- 9396648 TI - Outcomes analysis in asthma. AB - Physicians, patients, employers, managed care organizations, insurance companies, and government all want to know how different approaches to management of asthma are improving care. To this end, the field of outcomes analysis in asthma is playing a major role. Clinical, physiologic, humanistic, and economic outcomes are being assessed using different types of general and asthma-specific instruments. Historically, clinical and physiologic outcomes have been of most concern to clinicians. However, humanistic outcomes, such as health-related quality of life and patient satisfaction, shift the focus to the patient. Economic outcomes, especially cost-effectiveness, evaluate how to achieve the best outcomes at the lowest cost. These outcomes have been used to evaluate asthma intervention programs. Several large asthma outcomes research projects, which should define the future of outcomes analysis in asthma, are under way. PMID- 9396649 TI - Immunotherapy with allergens. AB - Allergen immunotherapy has been shown to be efficacious in numerous studies for the clinical indications of allergic asthma and rhinitis, as well as hymenoptera venom hypersensitivity. How allergen immunotherapy improves clinical symptoms is still not entirely clear. Decreases in specific IgE follow a complex cascade of effects: a shifting of the cytokine milieu from T(H)2 to T(H)1 predominance, with resultant decrease in interleukin 4, decreased recruitment and activation of eosinophils, and decreased proliferation of mast cells. Allergen exposure has a lessened ability to stimulate an inflammatory cell response, with decreased target organ hyperreactivity. Since allergen immunotherapy is not without risk, the decision needs to be made whether injection therapy is safe and provides benefit not achievable by medical management. The continued clarification of optimal allergen concentrations through careful studies of standardized extracts will allow better control of adverse events by limiting unnecessarily potent mixtures. PMID- 9396650 TI - Food allergy. AB - The evaluation of adverse reactions to foods involving abnormal immune responses to food allergens remains an important part of the practice of allergy and immunology. Approximately 5% of children younger than 3 years and 1.5% of the general population experience food allergic disorders, indicating that about 4 million Americans suffer from food allergies. The evaluation of adverse reactions to foods depends on a careful clinical history, diagnostic studies including appropriate skin testing or in vitro testing with food extracts, and/or endoscopy and biopsy. The mainstay of therapy remains avoidance of incriminated foods and education to deal with inadvertent exposures. Experience over the past decade suggests that the ready availability and early introduction of highly allergenic foods (eg, peanuts and nuts) into the diet will only increase the number of individuals suffering from hypersensitivity reactions to foods. Research has focused on the identification and characterization of allergenic proteins and the development of new therapeutic strategies, eg, plasmid DNA vaccines, to treat these disorders. PMID- 9396651 TI - Allergic reactions to drugs and biologic agents. AB - Drug allergies are adverse reactions resulting from immunologic responses to drugs or their metabolites. These reactions result in predictable patterns of organ-specific or systemic hypersensitivity that usually recur on subsequent exposure to the same drug. Although diagnostic testing for drug allergy is available for only a few drugs, protocols have been developed to assist in management of allergic reactions to many drugs and biologic agents. Other protocols assist in the management of patients who develop drug reactions while undergoing multiple drug therapy or those with a history of adverse drug reactions who again require treatment for the same condition. PMID- 9396652 TI - Allergic reactions to workplace allergens. AB - Allergic sensitization to workplace allergens can result in occupational asthma (OA), rhinitis, and dermatoses. Occupational asthma, accounting for 2% to 15% of all new cases of asthma, is caused by more than 240 reactive chemicals or natural proteins. Diisocyanates, used in urethane production and spray painting, are the leading causes of OA. Occupational asthma must be objectively confirmed by demonstrating significant decreases in lung function associated with exposure to a causative agent. An early diagnosis of OA followed by elimination of exposure to a causative agent may be curative and prevent progression to chronic asthma. In the last decade, protein allergens in natural rubber latex gloves have emerged as the leading cause of work-related cutaneous and respiratory allergic disorders in health care workers. In the workplace, occupational allergic contact dermatitis is almost always caused by chemicals, including nickel, chromates, and epoxy resins, whereas contact urticarial reactions are most often due to protein allergens. The primary treatment of occupational allergic disorders is strict avoidance of exposure to the inciting agent. PMID- 9396653 TI - Allergic and immunologic skin disorders. AB - The skin represents a unique immunologic organ poised to protect the host from invading organisms and environmental antigens. The skin is also an important target for a variety of allergic and autoimmune responses. Mast cells are key to the pathogenesis of urticaria, angioedema, and mastocytosis. Atopic dermatitis is the consequence of an immunoregulatory abnormality resulting in a skin-directed T helper type 2 response. Allergic contact dermatitis is an example of classic delayed type hypersensitivity. Circulating autoantibodies against the epidermis are a key mechanism by which bullous skin diseases occur. PMID- 9396654 TI - Immunologic aspects of lung diseases and cystic fibrosis. AB - Immunologic lung disorders are accompanied by an array of laboratory abnormalities, some of which contribute to disease pathogenesis. Allergic bronchopulmonary aspergillosis, which complicates asthma and cystic fibrosis, causes mucous plugging of airways, eosinophilic pneumonia, and bronchiolitis obliterans. Aspergillus fumigatus, growing saprophytically in bronchial mucus, is responsible for most cases, and prednisone, not antifungal agents, is the drug of choice because it controls the immunologic responses of the lung. In cystic fibrosis, epithelial surface fluid from the lung does not kill Pseudomonas aeruginosa, in part because antibodies to P aeruginosa are plentiful but ineffective in opsonizing bacteria. Neutrophil-derived elastase cleaves immunoglobulins and digests the C3b receptor on neutrophils, which limits phagocytosis of pathogens. In helminth infections and infestations, pulmonary and peripheral blood eosinophilia can be accompanied by increases in total and antiparasite IgE concentrations and generate T(H)2 CD4+ T-lymphocyte responses. Understanding the immunologic abnormalities of lung disorders may lead to more effective therapies. PMID- 9396655 TI - Autoimmune endocrine disease. AB - Autoimmune endocrine diseases are serious disorders that utilize immense health care resources and cause tremendous disability. They include type 1 diabetes mellitus, thyroiditis, Graves disease, Addison disease, and polyglandular syndromes. Analysis of the basis of autoimmune diseases has been aided by the application of new knowledge in immunologic physiology. Recent investigations using these techniques have revealed complicated disorders that have varied pathogenesis and complex genetic predispositions. While the mainstay of treatment for these diverse diseases remains the replacement of hormones produced by the damaged endocrine organ, investigations into the pathogenesis of these disorders provide hope for the development of specific therapeutic measures to block their pathologic basis. PMID- 9396656 TI - Immunologic aspects of renal disease. AB - The kidney can become involved in immune-mediated diseases through 3 mechanisms. It can be the primary target of antibody-mediated injury. Good-pasture syndrome, with antibodies directed at the glomerular basement membrane, is an example of this type. The kidney can be injured by immune complexes that are trapped in the kidney and cause local inflammation. Examples include systemic lupus erythematosus and IgA nephropathies. Finally, the kidney can be injured by immune responses initiated in other organs. In Wegener granulomatosis, inflammation begins in the airway but results in glomerulonephritis. Currently available therapies lack efficacy and specificity. New therapies based on pathophysiology are being developed in animal models. PMID- 9396657 TI - Immunopathogenesis of gastrointestinal and hepatobiliary diseases. AB - The largest lymphoid organ in the body is the gut and the gut-associated lymphoid tissue. The mucosal immune system faces many challenges in protecting the body from microbial invasion. Its chief function is to maintain a diverse population of mature lymphocytes capable of responding to foreign antigens. This task is accomplished with a variety of unique features that distinguish the mucosal from the systemic immune system. In addition, the mucosal immune system plays a role in inflammatory bowel disease, Whipple disease, autoimmune gastritis, Helicobacter pylori infection, immunoproliferative small intestinal disease, hepatitis A, B, C, D, E, F, and G, autoimmune hepatitis, primary biliary cirrhosis, progressive sclerosing cholangitis, and vanishing bile duct syndrome. PMID- 9396658 TI - Immunologic aspects of neurologic and neuromuscular diseases. AB - Inflammatory disorders of the nervous and neuromuscular system are not uncommon despite the fact that immune privilege exists in much of the nervous system. Common immune-mediated neurologic diseases include multiple sclerosis, myasthenia gravis, chronic inflammatory demyelinating polyneuropathy, and idiopathic polymyositis. Environmental, genetic, and immunologic factors have been postulated to be involved in the pathogenesis of these diseases, but much remains to be elucidated about the specific identity and relative contributions of these factors. Several new therapies have become available for these diseases in the past few years, and many others are under investigation. Strategies that enhance the normal tolerance mechanisms of the immune system are being developed. In particular, strategies to block T(H)1-type responses or enhance T(H)2/3-type responses have generated interest. PMID- 9396659 TI - Immunologic aspects of vasculitis and cardiovascular disease. AB - The immunologic cardiovascular diseases are a heterogeneous group of conditions. Many of these entities are associated with serious morbidity and mortality resulting from cardiac impairment, ischemic complications, or organ dysfunction, particularly of the kidneys, lung, and nervous system. The systemic nature of these conditions, coupled with vague symptoms and nonspecific initial physical findings, makes the differential diagnosis complicated. Research has identified specific mediators and primary origins in some conditions, with infections often being responsible. Immunosuppressive therapy, despite the potential complications, has improved the prognosis of some of the more serious immunologic cardiovascular diseases. Improvement in the treatment of immunologic cardiovascular diseases awaits identification of additional causes, improved definition of host factors that predispose an individual to develop these conditions, and better understanding of the immune dysregulation responsible for the progression of disease. The potential pathophysiologic role of immunologic mechanisms in common disorders such as congestive heart failure and atherosclerosis is intriguing and offers the possibility of novel therapies in these prevalent conditions. PMID- 9396660 TI - Tumor immunology. AB - Malignant tumors express antigens that may stimulate and serve as targets for antitumor immunity. Virally induced tumors usually contain integrated proviral genomes in theircellulargenomes and often express viral genome-encoded proteins that may stimulate specific host immune responses. Antigens unique to individual tumors that stimulate specific rejection of transplanted tumors have been demonstrated only in experimental animals. Other tumor antigens that potentially can stimulate immune responses are shared by different tumors. These include products of mutated or rearranged oncogenes or tumor-suppressor genes. Tumors may also overexpress tissue differentiation antigens or embryonic antigens, which also have the potential to be recognized by the immune system. The recent identification of tumor antigens recognized by cytotoxic T cells opens up new possibilities for constructing chemically defined antigens for specific immunotherapy. Treatment of malignant tumors in humans by immunologic approaches, although theoretically attractive, has not yet succeeded on a large scale. Important progress in immunotherapy of cancer is emerging with several different treatment modalities. PMID- 9396661 TI - Immunohematologic disorders. AB - Immunohematology encompasses a broad array of clinical disorders in which immune reactions are involved in the pathogenesis of hematologic diseases. Immune reactions can involve the formed elements of the blood, producing hemolytic anemia, thrombocytopenia, or neutropenia. Autoimmune phenomena and drug-induced reactions are the most common mechanisms. In newborns, maternal antibodies can cross the placenta and destroy red blood cells, platelets, or neutrophils. Immune reactions can also occur during transfusion of blood products, leading to hemolysis, febrile reactions, allergic reactions, and lung injury. The role of leukocytes and cytokines released during blood component storage in mediating febrile transfusion reactions has prompted the increased use of leukocyte-reduced components. Immune reactions can occur to soluble clotting factors and can produce bleeding or thrombosis. Finally, immunohematologic features of B-cell disorders are considered. PMID- 9396662 TI - Transplantation immunology. AB - The practice of clinical and experimental transplantation continues to evolve at a rapid pace. To appreciate the current transplant practices, it is first necessary to review transplant immunology in its proper context, ie, as a component of the complex series of events that promote the repair of damaged tissues. These processes are generally categorized as inflammation, immunity, and tissue repair/reinforcement. In general, there are 3 forms of graft rejection: hyperacute, acute, and chronic rejection. All 3 forms of graft rejection represent pathologic consequences of one or more of these repair-related processes. The various graft rejection responses also illustrate several complex immunologic principles that need to be considered. These include the definition of an alloantigen, the structure and function of major histocompatibility complex molecules, and the behavior of antigen-presenting cells and alloreactive T cells. This review combines these concepts and principles into a discussion of the 3 forms of graft rejection, each of which is addressed at the level of histopathology, pathobiology, incidence, and clinical strategies. PMID- 9396663 TI - Immunization. AB - Immunization is undergoing important changes, with improved vaccines replacing less immunogenic or less safe vaccines, new vaccines for common diseases such as chickenpox and hepatitis A infection, and improved immunization schedules. Immunization is also being transformed by basic work in molecular medicine. Vaccines made of DNA are being developed as a form of gene therapy that use the patient's own cellular machinery to make foreign proteins that stimulate an immune response. Currently immunization is used to protect patients prior to exposure to an infectious agent or during the incubation phase after exposure, but before disease has occurred. New technologies are being investigated to induce the immune system to fight infections that have already produced chronic disease such as acquired immunodeficiency syndrome and chronic hepatitis B virus infection. PMID- 9396664 TI - Immunopharmacology: immunomodulation and immunotherapy. AB - Immunopharmacology has changed dramatically over the past 25 years. Although a variety of traditional nonspecific immunosuppressive drug therapies are available for the treatment of autoimmune disease and organ transplantation rejection, with advances in cell biology and monoclonal antibody technology, a highly specific antibody can be engineered to cell surface determinants on immune cells or tumors or to neutralize inflammatory and immune mediators from an immune response. Many of these modalities are still in early phases of study for the treatment of autoimmune disease. In addition to therapies that suppress immune responses, advances in molecular biology have led to new agents and methods to enhance immune responses and correct immune deficits, such as growth factor replacement and cytokine therapies. Finally, gene therapy is a method for the long-term treatment of disorders in which a defective gene leads to disease. PMID- 9396665 TI - Outcome analysis and cost assessment in immunologic disorders. AB - A number of novel biologic agents are being introduced to replace, enhance, or modulate immune responses in medical illnesses. The use of these therapies has become crucial in treating some of these diseases, yet there is relatively little available information about their cost-effectiveness. Two examples are presented. Interferon gamma, used in chronic granulomatous disease, costs about $140 for a 100-microg vial; yearly costs average $21840 per patient. Study data estimated a 69% to 76% reduction in serious illness with interferon gamma treatment; a reduced incidence of infections could cover drug costs. Intravenous immunoglobulin is used lifelong in antibody deficiency and clearly reduces the number of serious illnesses. Projected savings derive from fewer hospital admissions and reduced organ damage, but infusion costs vary widely because of the prices charged for the drug and infusion services. PMID- 9396666 TI - Future trends in allergy and immunology. PMID- 9396667 TI - Allergy and immunology on the Internet. PMID- 9396668 TI - Acute sinusitis in the rabbit: a new rhinogenic model. AB - The objective of this study was to create a rhinogenic model of sinusitis using an implanted foreign body in the nasal cavity of the rabbit. The study design was a prospective controlled trial of the experimental design. In this model an obstructing foreign body was placed into the nasal cavity and then impregnated with pathogenic bacteria. This model was studied histologically using whole-mount techniques. Quantitative assessment of the degree of inflammation was made for the maxillary and ethmoid sinus and for overall effect for each animal. Bacteriologic study was performed in a limited number of animals. The results indicated that a significant infection develops in about half of animals. This peaks in intensity between 1 and 2 weeks after implantation and subsequently subsides with some evidence of long-term changes present after 4 weeks. It is concluded that this is a viable and perhaps preferable animal model to study sinusitis. Further investigation is necessary to completely characterize this model. PMID- 9396669 TI - Role of middle meatus aspiration culture in the diagnosis of chronic sinusitis. AB - Although empiric antibiotic therapy is often used for sinusitis, the emergence of antibiotic resistance has increased the failure rate of this approach. Culture directed therapy usually increases treatment success, but traditional antral puncture is often accompanied by poor patient and physician acceptance. Endoscopically directed middle meatal aspiration culture is increasingly used in this setting, but studies have not convincingly demonstrated the validity of this technique. Both endoscopic middle meatal and direct antral cultures were performed during endoscopic sinus surgery. Cytologic examination was performed to confirm the presence of inflammatory cells. When culture results were compared in 21 specimen pairs, exact correlation was found in 18 (85.7%). Based on this study, endoscopically directed middle meatus aspiration culture appears to be a valuable alternative to antral puncture for guiding organism-specific antibiotic therapy in sinusitis. PMID- 9396670 TI - Isolated sphenoid sinus disease: an analysis of 132 cases. AB - Solitary involvement of the sphenoid sinus is a relatively uncommon entity. A series of 132 patients with isolated sphenoid disease accumulated over a 22-year period is reported. A retrospective chart review was performed with special attention to the patients' presenting signs, symptoms, and radiographic findings. There were 80 patients with inflammatory disease, 38 with neoplasms, four with fibroosseous disorders, and 10 with traumatic and developmental lesions. The most common presenting symptom was headache, followed by visual changes and cranial nerve palsies. Cranial nerve abnormalities were encountered in 12% of the inflammatory cases, 60% of the benign tumors, and 57% of the malignant tumors. Radiographically, bone remodeling was associated with chronic inflammatory disease, especially mucoceles. Bone erosion was found principally with neoplastic disease, occurring rarely with mucoceles. Extension was associated with malignant tumors. PMID- 9396671 TI - Interaction between hypertension and diabetes mellitus in the pathogenesis of sensorineural hearing loss. AB - The purpose of this study is to support the hypothesis that diabetic end-organ damage of the cochlea is augmented in the setting of hypertension. A historical perspective reviewing the effects of diabetes and hypertension as causative factors in the development of sensorineural hearing loss, as well as the basic epidemiology and pathophysiology of the renal and vascular effects of diabetes and hypertension, is presented. The results of audiologic findings in insulin dependent diabetic patients, both normotensive and hypertensive, were analyzed and correlated with the results of animal studies to support the hypothesis that sensorineural hearing loss in patients and cochlear hair cell loss in animal studies result from the effects of hypertension in conjunction with insulin dependent diabetes mellitus. PMID- 9396672 TI - Nucleus 22 cochlear implantation results in postmeningitic deafness. AB - Cochlear implant surgery was performed on 13 patients with postmeningitic deafness (seven adults, six children). Two adults and two children (30.8%) had severe labyrinthitis ossificans requiring radical "drill-out." Five of 13 (38.5%) had some bone growth requiring partial drill-out, and four of 13 (30.8%) had normal insertion with no drill-out. Hearing results for patients with no bone growth were similar to nonmeningitic patients; three of four (75%) had open-set speech recognition. Performance of patients with total drill-out was poor; "auditory only" performance was limited to detection and pattern perception of speech, and no patients had open-set speech recognition. Results for patients with partial drill-out were similar to results in patients with no bone growth. Labyrinthitis ossificans not only presents surgical challenges to cochlear implantation but may also adversely affect hearing outcome. PMID- 9396673 TI - Cerebellopontine angle tumor outcomes in patients with hazardous occupations. AB - Cerebellopontine angle (CPA) tumor surgery can produce a spectrum of sensory and motor deficits that can alter a patient's lifestyle and occupation. An important consideration is whether patients can return to full occupational status especially when hazardous duties are involved. Outcome data in CPA tumor surgery usually report that most patients are able to return to preoperative function; however, whether these patients can return to hazardous duties is not specified. A retrospective study of 380 consecutive, operated CPA tumor patients was performed and 37 were identified who engaged in hazardous occupations including active military service, working at dangerous heights, piloting jet aircraft, aircraft navigating, and factory work with high-impact machinery. Overall, patients were able to resume full-time work in their previous occupations and 86% of patients reported that they were able to resume hazardous duties. CPA tumor surgery is compatible with continued full occupational duties after surgery, even for patients employed in hazardous situations. PMID- 9396674 TI - Incus and stapes footplate simulator. AB - Prosthesis placement in stapes surgery is difficult to master. Although temporal bone dissection is an important adjunct to operative experience and anatomic knowledge when training residents to perform this procedure, the high cost and scarcity of temporal bones available for teaching purposes limit their convenience as teaching tools. Therefore, to augment training of residents, the authors developed an inexpensive, easily constructed middle ear simulator made from a disposable drinking cup, toothpicks, and a tongue depressor. The device is used with a microscope and ear instruments to manipulate, place, and crimp stapes prostheses. Improved surgical skills can lead to better surgical results, and this middle ear simulator can help to improve otologic surgical skills by giving residents the opportunity to practice techniques necessary to master stapes surgery. PMID- 9396675 TI - Complications of gold weight eyelid implants for treatment of fifth and seventh nerve paralysis. AB - Complications occurred in six patients after gold weights were implanted into the upper eyelid tissues for fifth and seventh nerve palsies. These complications included implant infection without extrusion (in one patient); entropion with trichiasis and presumed inflammatory reaction to the gold weight material (in one patient); upper eyelid distortion and poor eyelid contour with corneal ulceration and scarring (in one patient); significant residual lagophthalmos with exposure keratitis (in one patient); and blepharoptosis obscuring the pupillary access (in two patients). Resolution of the complications required 1. implant removal in four of six patients without reinsertion of a second weight, 2. recession of the retractors of the upper eyelids with medial and lateral canthoplasty (in four patients), and 3. permanent tarsorrhaphy (in one patient). The authors conclude that complications may be minimized by careful preoperative determination of the optimum implant size, weight, and placement within the eyelid as well as meticulous attention to the surgical technique of implantation. The use of other eyelid protective procedures is often necessary to augment corneal protection especially in patients with combined fifth and seventh cranial nerve palsies. Endogenous implant infection without extrusion of the gold weight may be distinguished from presumed inflammation due to gold allergy by clinical response to antibiotics in the former and requirements of steroids or removal of the implant in the latter. PMID- 9396676 TI - Extended canine laryngeal preservation for transplantation. AB - The goal of successfully transplanting the larynx has motivated researchers since the 1960s. Early laryngeal transplant techniques limited the donor larynx to 45 minutes of ischemia. In this study, a method of prolonged laryngeal preservation is employed in three canines. In vivo cold laryngeal perfusion with University of Wisconsin Solution (UWS) was performed. The larynx was removed and placed into cold storage in 4 degrees C UWS. After 24 hours of storage, the same canines underwent laryngeal reimplantation. The animals were sacrificed 7 days after reimplantation. No evidence of necrosis or vascular insufficiency was identified histologically. The results indicate that canine larynges can be successfully reimplanted after 24 hours of preservation. Future studies will assess the application of this technique to laryngeal transplantation. PMID- 9396677 TI - Deep neck infections in children: a new approach to diagnosis and treatment. AB - Forty-seven children presented with the diagnosis of a deep neck infection- either cellulitis or abscess--between January 1991 and July 1996. Forty-four (94%) had contrast-enhanced computed tomography (CT) imaging consistent with this diagnosis. Three patients with no CT scan had confirmation of an abscess at surgical drainage. Parenteral antibiotics alone were effective in the treatment of 24 of 47 infections (51%): seven parapharyngeal, one retropharyngeal, and 16 combined. By CT scan these infections represented cellulitis in 17 of 24 (71%), an abscess in three of 24 (13%), and incomplete abscess in four of 24 (17%). The average duration of hospitalization for this group was 4.8 days, with symptomatic improvement usually seen within 24 hours. Surgical drainage was performed on 23 of 47 infections (49%): three parapharyngeal, 17 combined, and three of unknown specific location. In 22 of these 23 children (96%), transoral drainage of the abscess was used as the primary surgical approach. In 21 of these 22 (95%) there was complete resolution without complications or recurrence; one abscess required a subsequent external approach. CT scanning with contrast revealed that all deep neck infections were located medial (usually anteromedial) to the great vessels. Abscesses with volumes estimated to be greater than 2000 mm3 were more likely to undergo surgery, but these differences were not statistically significant. The use of contrast-enhanced CT scanning provides information regarding abscess size, location, and relative position of the great vessels for safe and successful transoral drainage. Thus we recommend CT-assisted transoral drainage for combined retropharyngeal/parapharyngeal abscesses and selected isolated parapharyngeal abscesses that do not respond to parenteral antibiotics. PMID- 9396678 TI - Characterization of smoking-induced nasopharyngeal lymphoid hyperplasia. AB - The frequency of smoking-induced nasopharyngeal lymphoid hyperplasia in heavy smokers and its potential clinical implications are still unknown. Precise criteria to differentiate this entity from other types of nasopharyngeal lymphoid hyperplasia are needed. A prospective clinicopathological study of smoking induced nasopharyngeal lymphoid hyperplasia was conducted in 17 heavy smokers. Ten nonsmoking patients, five of them with chronic sinusitis, three with adult onset adenoid hypertrophy, and two children with adenoidal hypertrophy served as a control group. Both in smokers and in nonsmokers, lymphocytic infiltration of the mucosa was characterized immunohistochemically as T cells. In smokers, semithin (1 micron) sections revealed deformed and migrating cytotoxic lymphocytes in the nasopharyngeal mucosa. The lymphocytes were attached to epithelial, ciliated, and goblet cells, resulting in cell damage. Transmission electron microscopy of biopsies from smokers revealed emperipolesis, characterized by mucosal invasion and epithelial cell damage by an unusual population of migrating T lymphocytes that penetrate them. These findings confirm a direct effect of smoking on the nasopharyngeal lymphoid tissue, which forms part of the immune system. It is concluded that the diagnostic evaluation and therapeutic approach of heavy smokers with otological and airway symptoms should be based on thorough endoscopic examination of the nasopharynx. When the diagnosis is not clear-cut, selective tele-endoscopic biopsy and electron microscopic examination are recommended. This entity should be added to the list of known clinical manifestations of the smoking habit. PMID- 9396679 TI - A clinical and histologic comparison of percutaneous dilational versus conventional surgical tracheostomy. AB - To directly compare percutaneous dilational tracheostomy (PDT) with conventional surgical tracheostomy, a prospective study was performed in 83 patients requiring tracheostomy for prolonged mechanical ventilation in the intensive care unit or after surgery for a large tumor in the upper respirodigestive tract. Median follow-up was 355 days after PDT and 338 days after conventional tracheostomy. The overall morbidity rate was significantly lower with PDT than with conventional tracheostomy (6.4% vs 36.1%; P < 0.001). Compared with conventional tracheostomy, PDT was also associated with a significantly lower incidence of postoperative bleeding (2.1% vs 13.9%; P < 0.05) and postoperative wound infection (0% vs 22.2%; P < 0.001). There were no clinical signs of laryngotracheal stenosis in either group. In conclusion, PDT is a simple, fast, safe bedside procedure that is associated with significantly lower morbidity than standard surgical tracheostomy. PMID- 9396680 TI - Tumors of the nasal columella treated by Mohs micrographic surgery. AB - Primary nasal columellar tumors are rare but can exhibit aggressive clinical behavior, often leading to devastating outcomes if these tumors are not completely removed. Eleven patients with nonmelanoma nasal columellar lesions were evaluated. All lesions were excised by obtaining tumor-free margins with Mohs micrographic surgery. Seven of eight cases with basal cell carcinoma have been tumor free for more than 5 years, the eighth case remains tumor free at 4 years. The primary squamous cell carcinoma lesions treated in this study have fared well; one patient has been tumor free for the past 4.3 years. No local recurrence or distant metastasis has occurred to date for all cases. We conclude that intervention with Mohs micrographic surgery for the treatment of early nasal columellar tumors may lead to improved outcomes. PMID- 9396681 TI - Fibrosing inflammatory pseudotumors of the central skull base. AB - The most important step in the differential diagnosis of mass lesions of the central skull base is to rule out malignant neoplasms. However, nonneoplastic lesions, such as infections or nonspecific inflammatory lesions of the skull base, can mimic malignant processes. In this study, the authors analyzed seven cases of nonneoplastic noninfectious mass-forming lesions involving the central skull base. In most cases, malignant processes were suspected at the initial phase of diagnostic work-up, but subsequent histologic examinations revealed that these lesions consisted of inflammatory cells and fibrosis without neoplastic cells. Common manifestations were pain and other neurological symptoms related to the involved anatomical sites. A variety of neurological dysfunctions of the cranial nerves not including the olfactory and spinal accessory nerves were observed. No patient developed separate lesions outside the head and neck region. After the pathologic diagnosis, most of the patients were treated with oral steroid therapy, with initial doses of prednisolone, 60 to 100 mg/d. It was difficult to relate responsiveness to steroid therapy with the histologic degree of sclerosis, fibrosis, or chronicity of the disease in these cases. Otolaryngologists should be aware of this disease when making treatment decisions for their patients with skull base lesions. PMID- 9396682 TI - Analysis of the mechanical behavior of the Nijdam voice prosthesis. AB - The valveless Nijdam prosthesis is a new voice prosthesis for laryngectomized patients using tracheoesophageal speech. An "umbrella-like hat" covers the esophageal side of the tracheoesophageal fistula and is deformed during speech by air pressure. To decrease pressure loss during speech, a good understanding of the mechanical behavior is essential. In the present study, the Finite Element Method (FEM), used in engineering to analyze the mechanical behavior of complex structures, was applied to analyze eight possible improvements of the Nijdam prosthesis. This study found that, during speech, deformation of hat and soft tissue occur. Distinct differences in the hat's deformation of the eight models also were found. It is concluded that complex structures like the Nijdam prosthesis can be analyzed by FEM. An optimal model was found to decrease pressure loss while stresses in the device remain safe. PMID- 9396684 TI - Evaluation of the bone resistance of the sphenoid and ethmoid sinuses. AB - Functional endoscopic sinus surgery can be associated with iatrogenic complications. Some anatomical structures have been identified this clinically as specific danger sites. The aim of this study was to confirm and to compare these clinical data with measurements of the bone resistance of the sphenoid and ethmoid walls and to point out regions that have increased bony fragility. Critical dynamometric evaluation of the resistance to breakage of these bony structures was made on 21 anatomic specimens. This study allowed the authors to localization of surgical complications, to demonstrate that other regions renowned for their fragility can be relatively robust, and to point out that robust sites can be dangerously fragile as a result of individual morphological variations. Results from this study provide a supplement to the guidelines for novice surgeons to use in identifying dangerous areas. PMID- 9396683 TI - Inhibitory effect of macrolides on interleukin-8 secretion from cultured human nasal epithelial cells. AB - The mechanism of macrolide therapy in chronic sinusitis patients is unclear. The authors studied the effect of macrolides on interleukin (IL)-8 secretion from cultured human nasal epithelial cells. Epithelial cells harvested from the nasal polyps of patients with chronic sinusitis were primary-cultured, and secreted IL 8 in culture media was measured by enzyme immunoassay. The cells secreted considerable amounts of IL-8 constitutively and in response to lipopolysaccharide. The secretion was significantly inhibited by 10(-5) M of erythromycin, clarithromycin, roxithromycin, and josamycin. 10(-6) M erythromycin still showed the inhibitory effect, whereas the same concentration of josamycin did not. These results indicate that macrolide antibiotics may act as an immunomodulator to reduce IL-8 in inflammatory sites and, at least partially, account for the clinically discrepant effects between 14- and 16-membered ring macrolides in long-term low-dose therapy for chronic sinusitis. PMID- 9396685 TI - Needle placement with transtympanic electrocochleography. AB - Electrocochleography (ECoG) is an objective, electrophysiologic test useful in the clinical diagnosis of endolymphatic hydrops, or Meniere's disease. The purpose of this study was to determine if the position of the needle, using transtympanic methodology, gives a variable SP/AP (summating potential/action potential) response. SP/AP ratios were obtained during routine tympanoplasty procedures. After the tympanic membrane remnant was removed using a lateral graft technique, precise needle placement was obtained at the medial and lateral round window niches, as well as on the promontory. SP/AP ratios were obtained in these three needle positions. There was no significant difference in the SP/AP ratio responses despite the location of needle placement. The use of transtympanic electrocochleography can give very good wave form morphology and consistent results. Therefore, if elevated SP/AP ratios do occur, they are thought to be due to a pathologic process of the ear and not needle placement. PMID- 9396686 TI - Mastoid obliteration and lining using the temporoparietal fascial flap. PMID- 9396687 TI - Correlation between preoperative computed tomography and preoperative findings in functional endoscopy sinus surgery. PMID- 9396689 TI - Clinical and epidemiologic features of Guillain-Barre syndrome. AB - Guillain-Barre syndrome (GBS) is defined clinically as a peripheral neuropathy causing limb weakness that progresses for up to 4 weeks before reaching a plateau. The symptoms may be caused by inflammatory demyelination, axonal degeneration, or both. GBS occurs throughout the world, with a median incidence of 1.3 cases/100,000 population (range, 0.4-4.0). Males are more commonly affected than females, and there are peaks in young adults and the elderly. There is no clear seasonal association in Western countries, although this may be because the most frequent antecedent events, respiratory and enteric infections, have opposite seasonality. The most frequently identified cause of GBS is Campylobacter jejuni infection, which has been identified in up to 41% of patients and is associated with more severe disease and prolonged disability. Summer epidemics of GBS occur among children and young adults in Northern China and are particularly likely to be associated with C. jejuni infection. PMID- 9396690 TI - Guillain-Barre syndrome: multifactorial mechanisms versus defined subgroups. AB - The clinical spectrum of Guillain-Barre syndrome (GBS) is summarized in relation to antecedent infections and anti-ganglioside antibodies. Associations exist between a pure motor form of GBS, diarrhea, Campylobacter jejuni infection, and anti-GM1 antibodies; between cranial nerve involvement and Miller Fisher syndrome, C. jejuni infection, and anti-GQ1b antibodies; and between variants, such as severe sensory involvement and cytomegalovirus infection. These three clinical variants are suggested to form the extremes of a continuous spectrum; they are discussed in relation to the more pathologically defined patterns of acute motor axonal neuropathy and acute motor-sensory axonal neuropathy. In particular, patients with a clinically pure motor variant of GBS, diarrhea, anti GM1 antibodies, or C. jejuni infection seem to respond better to early treatment with high-dose immunoglobulins than to plasma exchange. PMID- 9396691 TI - Epidemiologic and clinical features of Campylobacter jejuni infections. AB - Gram-negative bacteria of the genus Campylobacter and of related genera frequently colonize the gastrointestinal tracts of humans, other mammals, and birds. One organism, Campylobacter jejuni, has been recognized as an important human pathogen, usually causing a diarrheal illness. Infection is common throughout the world, but clinical and epidemiologic features differ in developed and developing countries. The high incidence of C. jejuni infections and their propensity to invade tissue and to induce inflammation are compatible with a role in the causation of Guillain-Barre syndrome. PMID- 9396692 TI - Microbiologic approaches for studying Campylobacter species in patients with Guillain-Barre syndrome. AB - Campylobacter jejuni is now considered to be the most common cause of bacterial diarrheal disease in the United States. Sufficient evidence exists to support the hypothesis that C. jejuni induces Guillain-Barre syndrome (GBS); however, many questions about the biology of the organism and host factors need to be answered. In order to study the role of C. jejuni and other Campylobacter species as a cause of GBS, isolates from patients with different forms of GBS and appropriate control populations must be obtained. To continue to study this association, research teams must have laboratory support for isolating and characterizing Campylobacter strains. This review summarizes current knowledge about the laboratory aspects of Campylobacter infection that may be pertinent to studies on GBS. PMID- 9396693 TI - Structure and conserved characteristics of Campylobacter jejuni lipopolysaccharides. AB - The lipid A component of Campylobacter jejuni lipopolysaccharides (LPSs) contains the same architectural principle as that found in other bacterial species; however, unlike the case with other bacterial species, the lipid A backbone of C. jejuni strains is composed of a phosphorylated beta(1'-6)-linked disaccharide containing 2,3-diamino-2,3-dideoxy-D-glucose and D-glucosamine as the major molecular species. Despite a backbone that differs structurally from that of classic enterobacterial lipid A, C. jejuni lipid A is antigenically similar to enterobacterial lipid A, and the respective LPSs have comparable endotoxic activities. Structural variability is greater in the core oligosaccharide than in lipid A of C. jejuni LPS. Nevertheless, the inner core oligosaccharides of C. jejuni strains share a common unique tetrasaccharide and also possess a trisaccharide that occurs in the inner core of other bacterial species. Ganglioside mimicry in the outer core is a common feature shared by a number of C. jejuni serotypes, but this mimicry is not conserved in all serotypes. PMID- 9396694 TI - Nonlipopolysaccharide surface antigens of Campylobacter species. AB - Among the protein antigens of Campylobacter species, flagellin, the subunit of the flagellar filament, is the best characterized. The motility imparted by this locomotory organelle is absolutely essential for Campylobacter organisms to colonize the gastrointestinal tract and to cause diarrheal disease. Flagellin is the immunodominant protein recognized during infection and has been suggested to be involved in the protective immune response. Campylobacter flagellins are glycosylated, which is an unusual posttranslational modification for prokaryotic proteins. Although the chemical structure of the glycosylated moiety is undetermined, the posttranslational modification includes sialic acid. The association of glycosylated flagellin with development of Guillain-Barre syndrome remains speculative, but the possibility of molecular mimicry between glycosylated flagellin and eukaryotic glycoproteins exists. PMID- 9396695 TI - Association between Campylobacter infection and Guillain-Barre syndrome. AB - Guillain-Barre syndrome (GBS), a neurologic disease that produces ascending paralysis, affects people all over the world. Acute infectious illnesses precede 50%-75% of the GBS cases. Although many infectious agents have been associated with GBS, the strongest documented association is with Campylobacter infection. The first line of evidence supporting Campylobacter infection as a trigger of GBS is anecdotal reports. The second line of evidence is serologic surveys, which have demonstrated that sera from GBS patients contain anti-Campylobacter jejuni antibodies, consistent with recent infection. Finally, culture studies have proven that a high proportion of GBS patients have C. jejuni in their stools at the time of onset of neurologic symptoms. Neurologic symptoms are more severe and more likely to be irreversible when GBS is preceded by C. jejuni infection. One of every 1058 Campylobacter infections results in GBS, and 1 of 158 Campylobacter type O:19 infections results in GBS. PMID- 9396696 TI - Campylobacter jejuni isolates from Japanese patients with Guillain-Barre syndrome. AB - Serologic evidence of recent Campylobacter jejuni infection was found in 92 (45%) of 205 Japanese patients with Guillain-Barre syndrome (GBS), and 49% of those 92 patients also had antibodies to GM1. Sixteen independent clinical isolates from GBS patients were serotyped: 12 belonged to Penner's heat-stable (HS) O serotype HS-19, 3 to HS-2, and 1 to HS-4. Of the patients whose C. jejuni isolates belonged to HS-19, 80% had elevated anti-GM1 antibodies. Although the correlation was significant between C. jejuni and GM1 antibody, anti-GM1 also was detected in 25% of patients without C. jejuni infection. Polymerase chain reaction-based restriction fragment length polymorphism analysis of an flaA gene showed that all HS-19 isolates, regardless of a GBS association, had an identical and distinguishable pattern, Cj-1, suggesting that HS-19:Cj-1 isolates are distinctive among C. jejuni isolates. Lectin typing showed that all GBS associated HS-19 isolates contained terminal beta-N-acetylglucosamine residues on their cell surface, but HS-19 isolates from patients with enteritis did not. PMID- 9396697 TI - Diversity of lipopolysaccharide structures in Campylobacter jejuni. AB - Immune blots of electrophoresed lipopolysaccharides extracted from 38 Campylobacter jejuni serostrains suggested the presence of O chains in 16 strains and their absence in 22. Structural analysis confirmed the presence of O chains in serostrains O:19, O:23, and O:36 and the absence of O chains in serostrains O:1, O:2, and O:3. The O:19 strain has O repeat units of beta-D-glucuronic acid amidated with 2-amino-2-deoxyglycerol and N-acetylglucosamine. The 0:36 O chain has four different but closely related repeat units that each consist of N acetylglucosamine, galactose, and a heptose that is varied in structure from one repeat unit to the next. The O:23 O chain has three different repeat units identical to three of O:36. Except for O:3, core oligosaccharides of strains with or without O chains contain sialic acid (Neu5Ac) in the terminal regions that in many cases mimic the structures of human gangliosides. Three neuropathic isolates were found to have a core terminal trisaccharide (Neu5Ac alpha2-->8Neu5Ac alpha2- >3Galbeta1) that was not found in nonneuropathic strains. PMID- 9396698 TI - Guillain-Barre syndrome in South Africa associated with Campylobacter jejuni O:41 strains. AB - Over a 20-month period, 3 adult and 6 pediatric patients were diagnosed with Guillain-Barre syndrome (GBS) at Groote Schuur and Red Cross Hospitals in Cape Town. All 9 GBS patients had Campylobacter jejuni biotype 2, serotype O:41 in their stools. C. jejuni infection was confirmed by ELISA testing of patient sera. Strains of this sero-biotype are rare: Only 12 such strains, including the GBS associated strains, were recognized among 776 Campylobacter strains isolated and identified at Red Cross Hospital from March 1994 to October 1995. This is the first known association of C. jejuni biotype 2, serotype O:41 with GBS. Patients infected with this Campylobacter strain had a particularly severe form of the infection, requiring hospitalization and ventilation much longer than GBS patients infected with other Campylobacter species and patients with Campylobacter-negative stools. The O:41 Campylobacter isolates from the GBS patients are identical by phenotypic, serologic, and molecular criteria, and they are clonal. PMID- 9396699 TI - Mechanisms of action of anti-GM1 and anti-GQ1b ganglioside antibodies in Guillain Barre syndrome. AB - Anti-GM1 and anti-GQ1b ganglioside antibodies are found in association with acute and chronic peripheral neuropathies, including Guillain-Barre syndrome. They are believed to arise as a result of molecular mimicry with immunogenic microbial polysaccharides. Although anti-ganglioside antibodies are suspected to play a causal role in neuropathy pathogenesis, the details of this have yet to be proven. The approach in this laboratory to solving this issue has been to generate anti-GM1 and anti-GQ1b monoclonal antibodies from peripheral blood lymphocytes of affected patients and to study their immunolocalization in peripheral nerve and their electrophysiologic effects in animal models in which peripheral nerve sites are exposed to anti-ganglioside antibodies. These data show that anti-ganglioside antibody-reactive epitopes are widely distributed in peripheral nerve and can cause electrophysiologic abnormalities in a variety of model systems; thus, these data support the view that anti-ganglioside antibody reactive epitopes may directly contribute to neuropathy pathogenesis. PMID- 9396700 TI - Molecular mimicry between gangliosides and lipopolysaccharides of Campylobacter jejuni isolated from patients with Guillain-Barre syndrome and Miller Fisher syndrome. AB - Some patients developed Guillain-Barre syndrome (GBS) after being given bovine gangliosides. Patients with GBS subsequent to Campylobacter jejuni enteritis frequently have IgG antibody to GM1 ganglioside. Miller Fisher syndrome (MFS), a variant of GBS, is associated with IgG antibody to GQ1b ganglioside. The existence of molecular mimicry between GM1 and lipopolysaccharide of C. jejuni isolated from a GBS patient and that between GQ1b and C. jejuni lipopolysaccharides from patients with MFS are shown herein. The molecular mimicry between infectious agents and gangliosides may function in the production of anti-ganglioside antibodies and the development of GBS and MFS. PMID- 9396701 TI - Acute immune polyneuropathies: correlations of serum antibodies to Campylobacter jejuni and Helicobacter pylori with anti-GM1 antibodies and clinical patterns of disease. AB - Antecedent Campylobacter jejuni infection, detected by serologic tests, has been implicated in some acute immune polyneuropathies (AIP). Antibodies to Helicobacter pylori, C. jejuni, and GM1 ganglioside were measured in sera from 35 Chinese patients with AIP. Anti-GM1 antibodies were found in 54% of C. jejuni seropositive, H. pylori-seronegative patients. In contrast, anti-GM1 antibodies were rare in sera that were either seropositive for both C. jejuni and H. pylori (P = .04) or seronegative for C. jejuni (P = .01). Motor axonal AIP was more common in the C. jejuni-seropositive, H. pylori-seronegative patients (82%) than in the bacterial antibody-negative group (38%). It was concluded that in AIP patients, C. jejuni-positive sera may be polyreactive, in that it may also react with H. pylori. In this situation, the specificity for either infection requires further validation. In contrast, sera with specific C. jejuni seropositivity are associated with both motor axonal AIP and selective serum IgG anti-GM1 antibodies. PMID- 9396702 TI - Anti-tubulin autoantibodies in acquired demyelinating polyneuropathies. AB - Selective high-titer anti-tubulin autoantibodies are associated with Guillain Barre syndrome and chronic inflammatory demyelinating polyneuropathy (CIDP). Low titer anti-tubulin autoantibodies are a normal component of some human sera. Analysis of 7 human sera with monoclonal anti-tubulin autoantibodies showed that the epitopes recognized by these antibodies are within central, conserved regions of tubulin. Sera from 3 patients with IgM monoclonal antibodies and CIDP reacted to an epitope spanning aa 301-314 of beta-tubulin, which has some sequence homology to several human viruses. Natural polyclonal anti-tubulin antibodies also bind to central regions of tubulin. In contrast, polyclonal tubulin antibodies induced in mice react to epitopes on the amino- or carboxy-terminal. Selective polyclonal anti-tubulin autoantibodies with low reactivity to other neural antigens are found in about one-half of patients with CIDP. PMID- 9396703 TI - P2-reactive T cells in inflammatory demyelination of the peripheral nerve. AB - Lewis rats immunized with P2 protein, a 14.5-kDa protein of the peripheral nerve myelin, develop experimental allergic neuritis, a paralytic disorder with clinical, histologic, and electrophysiologic features similar to those of human Guillain-Barre syndrome (GBS). T cells reactive to P2 protein or a peptide corresponding to 53-78 residues of the protein can transfer the disease to naive animals. The mechanisms by which these T cells induce demyelination are not well understood; however, they may induce inflammation and demyelination in the nerves by production of Th1 cytokines. Th2 cytokines may lead to suppression of the inflammation and eventual recovery. There is no conclusive evidence that P2 protein plays a role in the pathogenesis of GBS, with or without association with Campylobacter jejuni; however, studies of the immunopathogenesis of P2 protein induced experimental allergic neuritis are important for understanding the pathogenesis of inflammatory demyelination in the peripheral nerves, the hallmark of GBS. PMID- 9396704 TI - Insights into Campylobacter jejuni-induced Guillain-Barre syndrome from the Lewis rat model of experimental allergic neuritis. AB - Experimental allergic neuritis (EAN) is considered the in vivo model of Guillain Barre syndrome (GBS) and has been extensively studied in the Lewis rat. Both cellular and humoral components of the immune response are implicated in the inflammatory demyelination of peripheral nerves that characterizes EAN. The recognition of Campylobacter jejuni infection as a frequent antecedent event in GBS, and in particular its association with anti-ganglioside antibodies and a primary axonal neuropathy, has raised many questions about the specific disease mechanisms involved. While C. jejuni can produce an acute motor neuropathy in chickens, confirming the relationship between C. jejuni infection and acute neuropathy, no detailed information is available from this animal model. Insights from experimental studies relating to the effector phase of Lewis rat EAN that may be relevant to C. jejuni-induced GBS are discussed. PMID- 9396705 TI - Mechanisms of Schwann cell damage in inflammatory neuropathy. AB - Inflammatory demyelination of nerve in Guillain-Barre syndrome is triggered in most patients by prior infection with one of a series of organisms, including Campylobacter jejuni. The resulting inflammatory cascade, involving T cells, macrophages, complement, and cytokines, disrupts physiologic function of the peripheral nerve in part by targeting Schwann cells, the multipotential glial cells that synthesize multilamellar, compacted myelin and secrete growth factors. In vitro evidence suggests that the Schwann cell may itself be able to modulate the cascade by serving as an antigen-presenting cell and by producing cytokines and other acute-phase reactants. PMID- 9396706 TI - The role of cytokines in Schwann cell damage, protection, and repair. AB - Cytokines, proteins that are secreted by many cells, including inflammatory and glial cells, mediate interactions between cells, generally through paracrine and autocrine networks. Their effects are highly pleiotropic, with overlap of some activities. The pathogenesis of Guillain-Barre syndrome (GBS), especially the classic inflammatory demyelinating polyneuropathy form, seems to involve lymphocytes and macrophages, which are rich sources of cytokines. Macrophages likely have a role in the pathogenesis of the primarily axonal, less inflammatory forms of GBS. Cytokines appear to be involved in damage to Schwann cells, myelin, and axons, although the exact roles of the different cytokines is uncertain. There is increasing evidence that cytokines, including some proinflammatory cytokines that ordinarily cause damage, may also protect the cells of the peripheral nervous system and aid in its repair. The evolution of inflammatory and demyelinating disorders, including the degree of recovery, is probably dependent on the interactions of the different cytokines. PMID- 9396707 TI - Differential distribution of HLA alleles in two forms of Guillain-Barre syndrome. AB - Guillain-Barre syndrome in northern China occurs in two forms: acute inflammatory demyelinating polyneuropathy (AIDP) and acute motor axonal neuropathy (AMAN). AMAN and AIDP have an immunologic basis, and some cases are associated with preceding Campylobacter jejuni infection. The distribution of allelic forms of the histocompatibility genes HLA-DPB1, DQB1, DRB1, DRB3, DRB4, and DRB5 was examined by DNA-based technology in 34 control, 12 AIDP, and 31 AMAN cases. In AIDP patients, the DRB1*1301 allele showed a significant increase (18% vs. 0%, P = .055). In AMAN patients, alleles DRB1*1301-03 and DRB1*1312, taken collectively, were increased (19% vs. 0%, P = .009), but by itself, the DRB1*1301 allele was not increased, as in AIDP patients. With a larger number of persons, more definitive statements will be possible; however, the differential distribution of DR13 allelic forms between AIDP and AMAN cases may suggest that there are different immunologic mechanisms operating at the molecular level of these diseases. PMID- 9396708 TI - Vaccines against Campylobacter jejuni. AB - Campylobacter jejuni is one of the most common causes of diarrhea worldwide. The gastrointestinal manifestations of Campylobacter infection range from watery diarrhea to severe dysentery. Campylobacter infection has also been linked to the postinfectious sequelae of reactive arthritis and Guillain-Barre syndrome. Evidence from epidemiologic and volunteer studies suggests that development of a vaccine to prevent gastrointestinal disease and limit colonization is possible. Efforts to develop live attenuated or subunit vaccines are limited by the finite knowledge of Campylobacter pathogenesis and lack of a conserved protective antigen, respectively. An oral killed, whole-cell vaccine has been shown to be safe and effective in animal models and is currently being tested in phase I volunteer studies. PMID- 9396709 TI - Regulatory considerations for Campylobacter vaccine development. AB - The high, worldwide incidence of Campylobacter jejuni-associated diarrheal disease has recently prompted the development of anti-Campylobacter vaccines. However, the association of C. jejuni infections with subsequent development of Guillain-Barre syndrome has increased concerns from a pathogenesis standpoint and from a vaccine development and regulation standpoint. This brief overview describes the purpose and process of Food and Drug Administration review of vaccine products and highlights some important considerations pertinent to Campylobacter vaccine development. PMID- 9396710 TI - The economic burden of Campylobacter-associated Guillain-Barre syndrome. AB - Guillain-Barre syndrome (GBS) is an autoimmune disease characterized by acute neuromuscular paralysis. Of an estimated annual number of 2628-9575 US cases, 526 3830 are triggered by Campylobacter infection. Research objectives were to identify the lifetime consequences of GBS and, when possible, to quantify their economic burden. The cost-of-illness method was used to calculate annual societal resources spent on medical care and lost productivity due to illness or premature death from Campylobacter-associated GBS. Estimated total costs (in US$) of Campylobacter-associated GBS ($0.2-$1.8 billion) were added to previously estimated costs of campylobacteriosis ($1.3-$6.2 billion) for a total annual cost from Campylobacter of $1.5-$8.0 billion (1995 dollars). It is concluded that up to $8.0 billion in US human illness costs are spent annually because of Campylobacter infection. Economic evaluation of the other costs associated with GBS, such as physical and psychological costs, would increase these estimates. PMID- 9396711 TI - Workshop summary and recommendations regarding the development of Guillain-Barre syndrome following Campylobacter infection. PMID- 9396713 TI - Endothelin attenuates apoptosis in human smooth muscle cells. AB - An imbalance between proliferation and apoptosis is an important causal factor for disorders involving abnormal cell accumulation. Endothelin (ET)-1, a 21-amino acid peptide with mitogenic and vasoconstricting activities, not only acts as a mitogen, but also attenuates paclitaxel-induced apoptosis in smooth muscle cells. In both human pericardial and prostatic smooth muscle cells, addition of ET-1 reduced paclitaxel-induced DNA fragmentation and phosphatidylserine on the cell surface, two characteristics of apoptosis. By comparison, angiotensin II, another vasoactive peptide, did not have a significant effect on apoptosis. The effect of ET-1 was dose-dependent with an EC50 of 1 nM. These results suggest that ET is a potential survival factor for smooth muscle cells, and that altered activity of the ET system in disease states has potential to contribute to aberrant cell growth. PMID- 9396712 TI - Structure and physiological function of calpains. AB - For a long time now, two ubiquitously expressed mammalian calpain isoenzymes have been used to explore the structure and function of calpain. Although these two calpains, mu- and m-calpains, still attract intensive interest because of their unique characteristics, various distinct homologues to the protease domain of mu- and m-calpains have been identified in a variety of organisms. Some of these 'novel' calpain homologues are involved in important biological functions. For example, p94 (also called calpain 3), a mammalian calpain homologue predominantly expressed in skeletal muscle, is genetically proved to be responsible for limb girdle muscular dystrophy type 2A. Tra-3, a calpain homologue in nematodes, is involved in the sex determination cascade during early development. PalB, a key gene product involved in the alkaline adaptation of Aspergillus nidulans, is the first example of a calpain homologue present in fungi. These findings indicate various important functional roles for intracellular proteases belonging to the calpain superfamily. PMID- 9396714 TI - Molecular cloning and characterization of a nitrobenzylthioinosine-insensitive (ei) equilibrative nucleoside transporter from human placenta. AB - Mammalian equilibrative nucleoside transporters are typically divided into two classes, es and ei, based on their sensitivity or resistance respectively to inhibition by nitrobenzylthioinosine (NBMPR). Previously, we have reported the isolation of a cDNA clone encoding a prototypic es-type transporter, hENT1 (human equilibrative nucleoside transporter 1), from human placenta. We now report the molecular cloning and functional expression in Xenopus oocytes of a cDNA from the same tissue encoding a homologous ei-type transporter, which we designate hENT2. This 456-residue protein is 46% identical in amino acid sequence with hENT1 and corresponds to a full-length form of the delayed-early proliferative response gene product HNP36, a protein of unknown function previously cloned in a form bearing a sequence deletion. In addition to placenta, hENT2 is found in brain, heart and ovarian tissue. Like hENT1, hENT2 mediates saturable transport of the pyrimidine nucleoside uridine (Km 0.2+/-0.03 mM) and also transports the purine nucleoside adenosine. However, in contrast with hENT1, which is potently inhibited by NBMPR (Ki 2 nM), hENT2 is NBMPR-insensitive (IC50<1 microM). It is also much less sensitive to inhibition by the coronary vasoactive drugs dipyridamole and dilazep and to the lidoflazine analogue draflazine, properties that closely resemble those reported for classical ei-type transport in studies with intact cells. PMID- 9396715 TI - Nascent very-low-density lipoprotein triacylglycerol hydrolysis by lipoprotein lipase is inhibited by apolipoprotein E in a dose-dependent manner. AB - In the present study it was investigated whether apolipoprotein (apoE) can inhibit the lipoprotein lipase (LPL)-mediated hydrolysis of very-low-density lipoprotein (VLDL) triacylglycerols (TAGs). Previous studies have suggested such an inhibitory role for apoE by using as a substrate for LPL either plasma VLDL or artificial TAG emulsions. To mimic the in vivo situation more fully, we decided to investigate the effect of apoE on the LPL-mediated TAG hydrolysis by using VLDL from apoE-deficient mice that had been enriched with increasing amounts of apoE. Furthermore, since plasma VLDL isolated from apoE-deficient mice was relatively poor in TAGs and strongly enriched in cholesterol as compared with VLDL from wild-type mice, we used nascent VLDL obtained by liver perfusions. Nascent VLDL (d<1. 006) isolated from the perfusate of the apoE-deficient mouse liver was rich in TAGs. Addition of increasing amounts of apoE to apoE-deficient nascent VLDL effectively decreased TAG lipolysis as compared with that of apoE deficient nascent VLDL without the addition of apoE (63.1+/-6.3 and 20.8+/-1.8% of the control value at 2.7 microg and 29.6 microg of apoE/mg of TAG added respectively). Since, in vivo, LPL is attached to heparan sulphate proteoglycans (HSPG) at the endothelial matrix, we also performed lipolysis assays with LPL bound to HSPG in order to preserve the interaction of the lipoprotein particle with the HSPG-LPL complex. In this lipolysis system a concentration-dependent decrease in the TAG lipolysis was also observed with increasing amounts of apoE on nascent VLDL, although to a lesser extent than with LPL in solution (72.3+/ 3.6% and 56.6+/-1.7% of control value at 2.7 microg and 29.6 microg of apoE/mg TAGs added respectively). In conclusion, the enrichment of the VLDL particle with apoE decreases its suitability as a substrate for LPL in a dose-dependent manner. PMID- 9396717 TI - Nuclear localization domain of thyroid transcription factor-1 in respiratory epithelial cells. AB - Thyroid transcription factor-1 (TITF-1) is a homeodomain containing transcription factor that binds to and selectively activates the expression of genes in thyroid and pulmonary epithelial cells. TITF-1 plays a critical role in gene expression and in organogenesis of lung and thyroid. In the present work, epitope-tagged TITF-1 proteins were used to identify the regions of the TITF-1 polypeptide that mediate nuclear localization and transcriptional activity in human lung adenocarcinoma cells. A series of TITF-1-flag deletion mutants was generated and transfected into H441 cells to determine amino acid sequences involved in translocation to the nucleus. Transfection of the TITF-1-flag mutants demonstrated that a nuclear localization signal (NLS) sequence, located at the N terminus of the homeodomain, is critical for nuclear targeting. The NLS was essential but not sufficient for translocation of TITF-1 to the nucleus, since deletion of the homeodomain itself also blocked nuclear translocation in the presence of NLS. Deletion of the N-terminal transactivation domain of TITF-1 completely abolished its transcriptional activation on the human surfactant protein-B promoter, and deletion of the C-terminal domain partially reduced its stimulatory activity. Nuclear translocation of TITF-1 depends on both an NLS and the homeodomain of the polypeptide. Both C- and N-terminal regions of TITF-1 are involved in transactivation of surfactant protein B gene expression in pulmonary cells. PMID- 9396716 TI - Separate bisphosphatase domain of chicken liver 6-phosphofructo-2-kinase/fructose 2,6-bisphosphatase: the role of the C-terminal tail in modulating enzyme activity. AB - The separate bisphosphatase domain (amino acid residues 243-468) of the chicken liver bifunctional enzyme 6-phosphofructo-2-kinase-fructose-2,6-bisphosphatase was expressed in Escherichia coli and purified to homogeneity. The fructose-2, 6 bisphosphatase activity of the separate domain was 7-fold higher than that of the native bifunctional enzyme, and exhibited substrate inhibition characteristic of the native enzyme. The inhibition of the enzymes by fructose 2,6-bisphosphate could be overcome by Pi, glycerol 3-phosphate and GTP. Deletion of 30 amino acid residues from the C-terminus of the separate domain resulted in around a 5-fold increase in the Vmax of the bisphosphatase. Also, the truncated form was more accessible to chemical modification by diethyl pyrocarbonate and N ethylmaleimide, suggesting a more open structure than the wild-type form. In addition, the mutation of cysteine-389 to alanine increased bisphosphatase activity by 20% and the Km value for fructose 2,6-bisphosphate by 3-fold, and both the point mutation at cysteine-389 and the deletional mutation led to the predominantly insoluble expression of the enzyme. The results indicated that the C-terminal tail plays a role in modulating the enzyme activity and suggested that the difference in the C-terminal tail sequence is responsible for the difference in activity of the chicken and rat liver fructose-2,6-bisphosphatases. It is postulated that an interaction between the C-terminal tail and the active site might be present. PMID- 9396718 TI - Effect of aging on the chaperone-like function of human alpha-crystallin assessed by three methods. AB - alpha-Crystallin can function as a molecular chaperone by preventing unwanted interactions. This paper presents the effects of aging and cataract on the chaperone-like properties of alpha-crystallin from soluble fractions from the cortex and nucleus of human lenses by using three assays: enzyme inactivation and two turbidity experiments. The three methods complemented each other. There was no decrease with age of chaperone-like function of cortical alpha-low and alpha high crystallin. Nuclear alpha-low crystallin showed a decrease, whereas alpha high crystallin showed no age-related change but its protective effect was diminished. Results from the nucleus of 40-year-old cataractous lenses seemed similar to those for clear lenses of equivalent age, whereas 80-year-old cataractous lenses showed decreased chaperone-like behaviour. PMID- 9396719 TI - Integrin-dependent translocation of p160ROCK to cytoskeletal complex in thrombin stimulated human platelets. AB - p160(ROCK) is a protein serine/threonine kinase that binds to GTP-Rho and is activated by this binding. We have recently found that the expression of p160(ROCK) induces focal adhesions and stress fibres in HeLa cells, whereas a dominant-negative form of this kinase suppresses Rho-induced formation of these structures, suggesting that this kinase is a downstream target of Rho in this process [Ishizaki, Naito, Fujisawa, Maekawa, Watanabe, Saito and Narumiya (1997) FEBS Lett. 404, 118-124]. To find out the mode of action of p160(ROCK), we developed immunoblotting with an anti-p160(ROCK) antibody and investigated the subcellular localization of p160(ROCK) during platelet aggregation. In resting human platelets, more than 90% of p160(ROCK) was present in the Triton X-100 soluble fraction. When platelets were stimulated with thrombin, approx. 10% of p160(ROCK) was translocated to the Triton X-100-insoluble fraction. This translocation was detected as early as 20 s after stimulation and reached a maximum at 5 min; it was suppressed by the addition of EDTA or an Arg-Gly-Asp-Ser peptide (RGDS), both of which inhibit integrin alphaIIbbeta3-mediated platelet aggregation. Using [32P]Pi-loaded platelets, we found that p160(ROCK) was phosphorylated in response to stimulation by thrombin. This phosphorylation, however, was not affected by the addition of EDTA and RGDS. These results suggest that p160(ROCK) translocates to cytoskeleton in a manner dependent on integrin ligation and works in an early stage of cytoskeletal reorganization in thrombin stimulated platelets. PMID- 9396720 TI - Metal-ion-binding properties of synthetic conantokin-G. AB - The secondary structure of the synthetic 17-residue peptide, conantokin-G (con G), a gamma-carboxyglutamate-containing marine cone snail neuroactive protein, is altered from a random conformation to one containing a very high level (>70%) of alpha-helix on binding of multivalent cations. The proportion of alpha-helix formed correlated well with the size of the cation and ranged from a low of approx. 7% with large cations, such as Ba2+, to more than 70% with smaller cations, such as Mn2+, Mg2+ and Zn2+. The valency of the multivalent cation was not as important, since tervalent lanthanides (Eu3+, Gd3+ and Tb3+) of ionic radius 106-109 pm induced similar levels (50-60%) of helix to those induced by Ca2+ and Cd2+ (ionic radii 109 and 114 pm respectively). Although the correlation was not as tight, smaller cations of the same valency allowed the helical transition to occur at lower concentrations than the larger cations. The spectroscopic and spectrometric properties of some of these cations permitted a more detailed analysis of the molecular nature of the cation-con-G binding. EPR based titrations with Mn2+ provided a binding isotherm that was deconvoluted to a single class of 2-3 Mn2+ sites of average Kd 3.9 microM. This number of sites was similar to that for Ca2+ [Prorok, Warder, Blandl and Castellino (1996) Biochemistry 35, 16528-16534], but a much lower Kd was displayed with Mn2+. Determinations by 1H NMR of the longitudinal relaxation rates of the water protons in Mn2+/con-G solutions at different magnetic field strengths corresponding to the proton Langmuir frequencies of 24, 300 and 500 MHz permitted calculation of the hydration number of Mn2+ in the complex, which was found to be 1.0. This indicates that five of the six co-ordination sites of Mn2+ are occupied by peptide atoms, probably oxygens. Titrations of the changes in Tb3+ fluorescence as a result of its binding to con-G gave an EC50 of 58 microM, a value nearly identical with that obtained by titration of the change in helicity of the peptide as a function of Tb3+ concentration. This shows that the macroscopic binding of Tb3+ to con-G is directly responsible for the alteration in secondary structure of the peptide. Finally, Cd2+ was found to be an extremely suitable cation for an NMR-based investigation of the amino acid residues of apo con-G that are perturbed by cation binding. In a limited example of the results of this study, it was discovered that originally equivalent CH2(delta) protons of Arg13 became distinctly magnetically non-equivalent in the Cd2+-bound helical form of con-G. This indicates that Arg13 is situated in the helix in such a way that the mobility of its side chain is highly restricted. In conclusion, the data show that a variety of multivalent cations with measurable spectroscopic and spectrometric properties interact similarly with con-G and generate extensive alpha-helical conformation in this peptide. PMID- 9396721 TI - Differential regulation of types-1 and -3 inositol trisphosphate receptors by cytosolic Ca2+. AB - Biphasic regulation of inositol trisphosphate (IP3)-stimulated Ca2+ mobilization by cytosolic Ca2+ is believed to contribute to regenerative intracellular Ca2+ signals. Since cells typically express several IP3 receptor isoforms and the effects of cytosolic Ca2+ are not mediated by a single mechanism, it is important to resolve the properties of each receptor subtype. Full-length rat types-1 and 3 IP3 receptors were expressed in insect Sf9 cells at levels 10-40-fold higher than the endogenous receptors. The expressed receptors were glycosylated and assembled into tetramers, and binding of [3H]IP3 to each subtype was regulated by cytosolic Ca2+. The effects of increased [Ca2+] on native cerebellar and type-1 receptors expressed in Sf9 cells were indistinguishable. A maximally effective increase in [Ca2+] reversibly inhibited [3H]IP3 binding by approx. 50% by decreasing the number of IP3-binding sites (Bmax) without affecting their affinity for IP3. The effects of Ca2+ on type-3 receptors were more complex: increasing [Ca2+] first stimulated [3H]IP3 binding by increasing Bmax, and then inhibited it by causing a substantial decrease in the affinity of the receptor for IP3. The different effects of Ca2+ on the receptor subtypes were not a consequence of limitations in the availability of accessory proteins or of artifactual effects of Ca2+ on membrane structure. We conclude that Ca2+ can inhibit IP3 binding to types-1 and -3 IP3 receptors although by different mechanisms, and that IP3 binding to type-3 receptors is stimulated at intermediate [Ca2+]. A consequence of these differences is that, at resting cytosolic [Ca2+], type-3 receptors are more sensitive than type-1 receptors to IP3, but the situation reverses at higher cytosolic [Ca2+]. Such differences may be important in generating the spatially and temporally complex changes in cytosolic [Ca2+] evoked by receptors linked to IP3 formation. PMID- 9396722 TI - Alternative mRNA splicing of 3'-terminal exons generates ascorbate peroxidase isoenzymes in spinach (Spinacia oleracea) chloroplasts. AB - We have isolated two cDNA clones encoding spinach (Spinacia oleracea) stromal and thylakoid-bound ascorbate peroxidase isoenzymes [Ishikawa, Sakai, Yoshimura, Takeda and Shigeoka (1996) FEBS Lett. 384, 289-293]. The gene (ApxII) encoding both chloroplastic ascorbate peroxidase isoenzymes was isolated and the organization of the gene was determined. Alignment between the cDNAs and the gene for chloroplastic ascorbate peroxidase isoenzymes indicates that both enzymes arise from a common pre-mRNA by alternative splicing of two 3'-terminal exons. Genomic Southern-blot analysis supported this finding. The gene spanned nearly 8.5 kbp and contained 13 exons split by 12 introns. The penultimate exon 12 (residues 7376-7530) for the stromal ascorbate peroxidase mRNA consisted of one codon for Asp365 before the TAA termination codon, and the entire 3'-untranslated region, including a potential polyadenylation signal (AATAAA). The final exon 13 (residues 7545-7756) for the thylakoid-bound ascorbate peroxidase mRNA consisted of the corresponding coding sequence of the hydrophobic C-terminal region, the TGA termination codon and the entire 3'-untranslated region, including a potential polyadenylation signal (AATATA). Both exons were interrupted by a 14 bp non-coding sequence. Northern-blot and reverse transcription-PCR analysis showed that the transcripts for stromal and thylakoid-bound ascorbate peroxidase are present in spinach leaves. PMID- 9396723 TI - Mechanism of the antimycin A-mediated enhancement of t-butylhydroperoxide-induced single-strand breakage in DNA. AB - Inhibitors of complex III increased the DNA strand scission induced by t butylhydroperoxide (tB-OOH) and cumene hydroperoxide but did not affect DNA damage induced by H2O2. The hypothesis that these effects are selectively linked to inhibition of the electron transport from cytochrome b to cytochrome c1 is validated by the following observations: (1) two complex III inhibitors, antimycin A and 2-heptyl-4-hydroxyquinoline N-oxide, enhanced the tB-OOH-induced DNA cleavage over the same concentration range as that in which inhibition of oxygen consumption was observed; (2) the complex III inhibitor-mediated enhancement of tB-OOH-induced DNA damage was abolished by the complex I inhibitor rotenone or by glucose omission, and (3) the enhancing effects of antimycin A were not observed in respiration-deficient cells. The mechanism whereby the complex III inhibitors potentiate DNA cleavage promoted by tB-OOH was subsequently investigated with intact as well as permeabilized cells. H2O2, produced at the level of mitochondria via a Ca2+-dependent process, was found to account for the enhancing effects of antimycin A. PMID- 9396724 TI - Mammalian cells express two differently localized Bag-1 isoforms generated by alternative translation initiation. AB - The Bcl-2 oncoprotein is a key regulator of apoptosis and the Bag-1 protein interacts with Bcl-2 and cooperates with Bcl-2 to suppress apoptosis. The human Bag-1 cDNA is essentially identical with a previously described cDNA encoding RAP46, which interacts with activated steroid hormone receptors. However, there is considerable confusion over the structure of Bag-1/RAP46 proteins and their relationship to endogenous Bag-1 proteins. Here we have characterized Bag-1 expression in mammalian cells. We demonstrate that, in addition to the previously identified 32 kDa murine and 36 kDa human Bag-1 proteins, cells express a second 50 kDa Bag-1 isoform. In some murine cell lines p50 is expressed at the same level as p32 Bag-1, and p50 and p32 Bag-1 proteins have distinct subcellular localizations, suggesting that they are functionally distinct. The published mouse Bag-1 cDNA is partial, and sequencing of additional murine Bag-1 RNA 5' sequences demonstrated that human and murine Bag-1 cDNAs contain longer open reading frames than originally suspected. We determined which open reading frames gave rise to the Bag-1 isoforms in human cells. Surprisingly, translation of neither protein initiated at the first in-frame methionine, and cells do not express Bag-1/RAP46 proteins with the previously proposed structures; p50 Bag-1 initiates at an upstream CUG codon, whereas p36 Bag-1 initiates at a downstream AUG codon. Therefore, cells express two differently localized Bag-1 isoforms generated by alternative translation initiation, and Bag-1 proteins may play a dual role in regulating apoptosis and steroid hormone-dependent transcription. PMID- 9396725 TI - Reconstitution, morphology and crystallization of a fatty acid beta-oxidation multienzyme complex from Pseudomonas fragi. AB - The fatty acid beta-oxidation multienzyme complex from Pseudomonas fragi, HDT, exhibits predominantly the three enzymic activities of 2-enoyl-CoA hydratase (EC 4.2.1.17), 3-hydroxyacyl-CoA dehydrogenase (EC 1.1.1.35) and 3-oxoacyl-CoA thiolase (EC 2.3.1.16). The HDT complex is encoded by the faoAB operon, consisting of the faoA and faoB genes that encode two individual constituents, the alpha-subunit and the beta-subunit. We have constructed Escherichia coli overexpression systems for the faoAB gene product (coexpression of the alpha- and beta-subunits), the alpha-subunit alone and the beta-subunit alone, and have purified the three respective products. Gel-filtration analysis revealed that the faoAB gene product forms a heterotetrameric structure, alpha2beta2, identical with the native HDT oligomeric state from P. fragi, whereas the alpha-subunit and beta-subunit individually form dimers. Electron microscopy demonstrated that each protein morphologically adopts the above oligomeric structures. The HDT complex, reconstituted in vitro from the isolated alpha- and beta-subunits, exhibits the three original enzymic activities and yields the same crystal as those from the native enzyme. CD measurements indicated that the alpha- and beta-dimers hardly alter their global conformations upon the formation of the HDT complex. Interestingly, the beta-dimer alone does not exhibit 3-oxoacyl-CoA thiolase activity, whereas the alpha-dimer alone exhibits both the 2-enoyl-CoA hydratase and 3-hydroxyacyl-CoA dehydrogenase activities. These results suggest that the contact between the alpha- and beta-subunits is essential for the thiolase activity. We have identified several structurally important proteolytic sites within each subunit, which are protected in the intact heterotetrameric molecule. These findings allow the possible location of the interface between the two subunits, which should be crucial for the exhibition of thiolase activity. PMID- 9396726 TI - Characterization of a Drosophila phosphorylation-dependent nuclear-localization signal-binding protein. AB - A 94 kDa nuclear-localization-signal (NLS)-binding protein was purified from Drosophila embryos. The NLS of the simian-virus-40 T-antigen is specifically bound by the dephosphorylated form of the protein. After phosphorylation, the affinity of the protein for the NLS is sharply decreased. In the dephosphorylated form, p94 (protein of 94 kDa) is the major NLS-binding protein in Drosophila embryos. Immunoprecipitation confirmed the ATP-dependent phosphorylation of p94, and co-precipitation of two additional phosphorylated proteins, indicated that the NLS-binding protein is part of a larger complex in Drosophila embryos. In agreement with the immunoprecipitation results, cross-linking experiments demonstrated the interaction of p94 with three additional proteins. These protein protein interactions were also phosphorylation-dependent. PMID- 9396727 TI - Haem precursor delta-aminolaevulinic acid induces activation of the cytosolic iron regulatory protein 1. AB - Control of cellular iron homoeostasis is performed by iron regulatory protein 1 (IRP1) through post-transcriptional modifications. This protein is sensitive to intracellular iron availability, being activated at low iron levels and inactivated at high iron levels, conditions that signal the increased expression of the transferrin receptor or of ferritin respectively. IRP1 is known to be activated by some oxidants such as H2O2 and NO. delta-Aminolaevulinic acid (ALA), previously found to produce reactive oxygen species and a carbon-centred radical, to release iron from ferritin, and to increase rat liver and brain non-haem iron and ferritin, was investigated for its effects on IRP1 activity in cultured hamster pulmonary fibroblasts. We have found that 1-2 mM ALA produced a 2-3-fold activation of IRP. On incubation with 1-4 mM succinylacetone methyl ester, a potent ALA dehydratase inhibitor, a 3-4-fold activation of the protein was observed, accompanied by a 40% increase in the intracellular ALA concentration. When cells were incubated in the presence of ALA or succinylacetone methyl ester, N-acetylcysteine inhibited IRP1 activation, suggesting that the observed effect is mediated by an oxidative process. We surmise that ALA-induced IRP1 activation might act as a co-sensor of iron homoeostasis. PMID- 9396728 TI - Covalent complex of microperoxidase with a 21-residue synthetic peptide as a maquette for low-molecular-mass redox proteins. AB - Here we report the structural and functional characterization of a covalent complex (MKP) obtained by cross-linking microperoxidase (Mp), the haem undecapeptide obtained by the peptic digestion of cytochrome c, with a 21-residue synthetic peptide (P21) analogous to the S-peptide of the RNase A. The covalent complex has been prepared by introducing a disulphide bond between Cys-1 of P21 and Lys-13 of Mp, previously modified with a thiol-containing reagent. On formation of the complex (which is a monomer), the helical content of P21 increases significantly. The results obtained indicate that His-13 of P21 co ordinates to the sixth co-ordination position of the haem iron, thus leading to the formation of a complex characterized by an equilibrium between an 'open' and a 'closed' structure, as confirmed by molecular dynamics simulations. Under acidic pH conditions, where His-13 of P21 is loosely bound to the haem iron ('open' conformation), MKP displays appreciable, quasi-reversible electrochemical activity; in contrast, at neutral pH ('closed' conformation) electrochemical behaviour is negligible, indicating that P21 interferes with the electron transfer properties typical of Mp. On the whole, MKP is a suitable starting material for building a miniature haem system, with interesting potential for application to biosensor technology. PMID- 9396729 TI - Dependence of the anti-chaperone activity of protein disulphide isomerase on its chaperone activity. AB - Protein disulphide isomerase (PDI) shows chaperone and anti-chaperone activities in assisting refolding of denatured and reduced lysozyme in redox Hepes buffer, but only chaperone activity in phosphate buffer and redox Hepes buffer containing 0.1 M NaCl. In non-redox Hepes buffer its anti-chaperone activity is very weak. PDI displays its anti-chaperone activity only for those substrates showing relatively low aggregation during refolding, and is strongly dependent on refolding conditions, of which ionic strength appears to be an important factor. The S-methylated PDI, fully active as a chaperone but devoid of isomerase activity, by itself shows only anti-chaperone activity, but reinforces rather than suppresses the chaperone activity of native PDI in the refolding of lysozyme. A fragment of PDI with the C-terminal peptide-binding sequence removed and devoid of chaperone activity does not show anti-chaperone activity in lysozyme refolding. It appears that the anti-chaperone activity of PDI is dependent on its chaperone activity. PMID- 9396730 TI - Excess putrescine accumulation inhibits the formation of modified eukaryotic initiation factor 5A (eIF-5A) and induces apoptosis. AB - DH23A cells, an alpha-difluoromethylornithine-resistant variant of the parental hepatoma tissue culture cells, express high levels of stable ornithine decarboxylase. Aberrantly high expression of ornithine decarboxylase results in a large accumulation of endogenous putrescine and increased apoptosis in DH23A cells when alpha-difluoromethylornithine is removed from the culture. Treatment of DH23A cells with exogenous putrescine in the presence of alpha difluoromethylornithine mimics the effect of drug removal, suggesting that putrescine is a causative agent or trigger of apoptosis. Accumulation of excess intracellular putrescine inhibits the formation of hypusine in vivo, a reaction that proceeds by the transfer of the butylamine moiety of spermidine to a lysine residue in eukaryotic initiation factor 5A (eIF-5A). Treatment of DH23A cells with diaminoheptane, a competitive inhibitor of the post-translational modification of eIF-5A, causes both the suppression of eIF-5A modification in vivo and induction of apoptosis. These data support the hypothesis that rapid degradation of ornithine decarboxylase is a protective mechanism to avoid cell toxicity from putrescine accumulation. Further, these data suggest that suppression of modified eIF-5A formation is one mechanism by which cells may be induced to undergo apoptosis. PMID- 9396732 TI - Purification of feline lysosomal alpha-mannosidase, determination of its cDNA sequence and identification of a mutation causing alpha-mannosidosis in Persian cats. AB - alpha-Mannosidosis is a lysosomal storage disorder that is caused by the deficiency of lysosomal alpha-mannosidase. Feline alpha-mannosidosis is a well characterized animal model used for studying pathological and therapeutic aspects of lysosomal storage disorders. We here report the purification of feline liver lysosomal alpha-mannosidase and determination of its cDNA sequence. The active enzyme consisted of three polypeptides, with molecular masses of 72, 41 and 12 kDa, joined by non-covalent forces. The cDNA sequence of feline lysosomal alpha mannosidase was determined from reverse transcriptase PCR products obtained from skin fibroblast mRNA. The deduced amino acid sequence contained the N-terminal sequences of the 72 and 41 kDa peptides. This indicated that the enzyme is synthesized as a single-chain precursor with a putative signal peptide of 50 amino acids followed by a polypeptide chain of 957 amino acids, which is cleaved into the three polypeptides of the mature enzyme. The deduced amino acid sequence was 81.1 and 83.2% identical with the human and bovine lysosomal alpha mannosidases sequences respectively. A 4 bp deletion was identified in an alpha mannosidosis-affected Persian cat by DNA sequencing of reverse transcriptase PCR products. The deletion resulted in a frame shift from codon 583 and premature termination at codon 645. No lysosomal alpha-mannosidase activity could be detected in the liver of this cat. A domestic long-haired cat expressing a milder alpha-mannosidosis phenotype than the Persian cat had a lysosomal alpha mannosidase activity of 2% of normal. This domestic long-haired cat did not possess the 4 bp deletion, proving molecular heterogeneity for feline alpha mannosidosis. PMID- 9396731 TI - Mechanism of acylphosphatase inactivation by Woodward's reagent K. AB - The organ common-type (CT) isoenzyme of acylphosphatase is inactivated by Woodward's reagent K (WRK) (N-ethyl-5-phenylisoxazolium-3'-sulphonate) at pH6.0. The inactivation reaction follows apparent pseudo first-order kinetics. The dependence of the reciprocal of the pseudo first-order kinetic constant (kobs) on the reciprocal WRK concentration reveals saturation kinetics, suggesting that the WRK forms a reversible complex with the enzyme before causing inactivation. Competitive inhibitors, such as inorganic phosphate and ATP, protect the enzyme from WRK inactivation, suggesting that this reagent acts at or near to the enzyme active site. The reagent-enzyme adduct, which elicits a strong absorption band with lambdamax at 346 nm, was separated from unreacted enzyme by reverse phase HPLC and the modified protein was cleaved with endoproteinase Glu-C to produce fragments. The HPLC fractionation gave two reagent-labelled peptides (peak 1 and peak 2) that were analysed by ion-spray MS and sequenced. The former is VFFRKHTQAE (residues 20-29 of human CT acylphosphatase) and the latter IFGKVQGVFFRKHTQAE (residues 13-29). MS demonstrated that both peptides are WRK adducts. A fragment ion with m/z of 1171, which is present in the mass spectrum of peak 1, has been identified as a WRK adduct of the peptide fragment 20-26. The lambdamax at 346 nm of WRK adduct suggests that the modified residue is His-25. Five recombinant enzymes mutated in residues included in the 20-29 polypeptide stretch have been produced. Analysis of their reactivities with WRK demonstrates that His-25 is the molecular target of the reagent as its modification causes the inactivation of the enzyme. Since both His-25-->Gln and His-25-->Phe mutants maintain high catalytic activity, we suggest that the observed enzyme inactivation is caused by the reagent (covalently bound to His-25), which shields the active site. PMID- 9396733 TI - Human endothelin-converting enzyme (ECE-1): three isoforms with distinct subcellular localizations. AB - Endothelin-converting enzyme 1 (ECE-1) is a membrane-bound metalloprotease that catalyses the conversion of inactive big endothelins into active endothelins. Two different isoforms (ECE-1a and ECE-1b) have previously been identified for human ECE-1. In the present study we have cloned a novel human ECE-1 isoform, termed ECE-1c, and have thus shown for the first time the existence of three distinct ECE-1 isoforms. The three isoforms differ only in their N-terminal regions and are derived from a single gene through the use of alternative promoters. Ribonuclease protection experiments revealed that, although the relative levels of the three isoform mRNA species vary between human tissues, ECE-1c mRNA is generally the predominant isoform messenger. Immunofluorescence microscopy analysis showed distinct subcellular localizations for the three isoforms: whereas ECE-1a and ECE-1c are localized at the cell surface, ECE-1b was found to be intracellular and showed significant co-localization with a marker protein for the trans-Golgi network. We determined that the three isoforms have similar kinetic rate constants (Km, kcat and Vmax) for the processing of big endothelin 1 and that the big endothelin isoforms 1, 2 and 3 are cleaved with similar relative velocities of 1.0:0.1:0.1 by the three isoenzymes. PMID- 9396734 TI - Structure of the mouse leukaemia inhibitory factor receptor gene: regulated expression of mRNA encoding a soluble receptor isoform from an alternative 5' untranslated region. AB - The low-affinity leukaemia inhibitory factor receptor (LIF-R) is a component of cell-surface receptor complexes for the multifunctional cytokines leukaemia inhibitory factor, ciliary neurotrophic factor, oncostatin M and cardiotrophin-1. Both soluble and transmembrane forms of the protein have been described and several LIF-R mRNAs have been reported previously. In order to determine the coding potential of LIF-R mRNAs we have isolated and characterized the mouse LIF R gene. mRNA encoding soluble LIF-R (sLIF-R) is formed by inclusion of an exon in which polyadenylation signals are provided by a B2 repeat. This exon is located centrally within the LIF-R gene but is excluded from the transmembrane LIF-R mRNA by alternative splicing. The transmembrane receptor is encoded by 19 exons distributed over 38 kb. Two distinct 5' non-coding exons have been identified, indicating the existence of alternative promoters. One of these is G/C rich and possesses a consensus initiator sequence as well as potential Sp1 binding sites. Expression of exon 1 from this promoter occurs in a wide variety of tissues, whereas expression of the alternative 5' untranslated region (exon 1a) is normally restricted to liver, the principal source of sLIF-R. During pregnancy expression of exon 1a becomes detectable also in the uterus. Expression of exon 1a increases dramatically during gestation and is accompanied by a similar quantitative rise in expression of sLIF-R mRNA. These findings establish that expression of LIF-R is under complex transcriptional control and indicate that regulated expression of the soluble cytokine receptor isoform may be due principally to an increase in the activity of a dedicated promoter. PMID- 9396735 TI - Selective labelling of cell-surface polyamine-binding proteins on leukaemic and solid-tumour cell types using a new polyamine photoprobe. AB - Polyamine transport is an active process which contributes to the regulation and maintenance of intracellular polyamine pools. Although the biochemical properties of polyamine transport in mammalian cells have been extensively studied, attempts to isolate and characterize the actual protein(s) have met with limited success. As one approach, photoaffinity labelling of cell surface proteins using a polyamine-conjugated photoprobe may lead to the identification of polyamine binding proteins (pbps) associated with the transport apparatus and/or other regulatory responses. In a previous study [Felschow, MacDiarmid, Bardos, Wu, Woster and Porter (1995) J. Biol. Chem. 270, 28705-28711], we demonstrated that the photoprobes N4-ASA-spermidine and N1-ASA-norspermine [where the ASA (azidosalicylamidoethyl) group represents the photoreactive moiety] competed effectively with polyamines for transport and selectively labelled two major pbps at 118 and 50 kDa on the surface of murine and human leukaemia cells. In the present study, a new and more potent polyamine-conjugated photoprobe, N1-ASA spermine, has been synthesized and used to develop a method based on detergent lysis for identifying putative cell-surface pbps on solid-tumour cell types. Transport kinetic assays showed that the new photoprobe competed with spermidine uptake with an apparent Ki of 1 microM, a value 20-50-fold lower than those of earlier probes. In L1210 cells, the new probe identified pbp50 and pbp118 thus reaffirming their identity as pbps. Two new bands were also detected. In A549 human lung adenocarcinoma cells, N1-ASA-spermine identified pbps at 39, 62, 73 and 130 kDa, the latter believed to be a size variant of pbp118. The presence of pbp130/118 in two very different cell types suggests the generality of the protein among mammalian cell types as well as its importance for further study. The high affinity of the photoprobe for the polyamine-transport system strongly suggests that at least some of the identified pbps may be associated with that function. PMID- 9396736 TI - Conformational changes of P-glycoprotein by nucleotide binding. AB - P-glycoprotein (Pgp) is a membrane protein that transports chemotherapeutic drugs, causing multidrug resistance in human cancer cells. Pgp is a member of the ATP-binding cassette superfamily and functions as a transport ATPase. It has been suggested that the conformation of Pgp changes in the catalytic cycle. In this study, we tested this hypothesis by using limited proteolysis as a tool to detect different conformational states trapped by binding of nucleotide ligands and inhibitors. Pgp has high basal ATPase activity; that is, ATP hydrolysis by Pgp is not rigidly associated with drug transport. This activity provides a convenient method for studying the conformational change of Pgp induced by nucleotide ligands, in the absence of drug substrates which may generate complications due to their own binding. Inside-out membrane vesicles containing human Pgp were isolated from multidrug-resistant SKOV/VLB cells and treated with trypsin in the absence or presence of MgATP, Mg-adenosine 5'-[beta,gamma-imido]triphosphate (Mg p[NH]ppA) and MgADP. Changes in the proteolysis profile of Pgp owing to binding of nucleotides were used to indicate the conformational changes in Pgp. We found that generation of tryptic fragments, including the loop linking transmembrane (TM) regions TM8 and TM9 of Pgp, were stimulated by the binding of Mg-p[NH]ppA, MgATP and MgADP, indicating that the Pgp conformation was changed by the binding of these nucleotides. The effects of nucleotides on Pgp conformation are directly associated with the binding and/or hydrolysis of these ligands. Four conformational states of Pgp were stabilized under different conditions with various ligands and inhibitors. We propose that cycling through these four states couples the Pgp-mediated MgATP hydrolysis to drug transport. PMID- 9396737 TI - Glycosylphosphatidylinositols of Plasmodium chabaudi chabaudi: a basis for the study of malarial glycolipid toxins in a rodent model. AB - Free and protein-bound glycosylphosphatidylinositols (GPIs) of the blood stages of the rodent malarial parasite Plasmodium chabaudi chabaudi AS were identified and characterized. TLC analysis of material extracted by organic solvents from metabolically labelled parasites revealed a distinct set of glycolipids. These glycolipids were identified as GPIs by specific chemical and enzymic treatments and by structural analysis of their glycan and hydrophobic parts. These analyses revealed that P.c.chabaudi AS synthesizes a set of GPI-biosynthesis intermediates and two potential GPI-anchor precursors exhibiting the following structures: ethanolamine-phosphate [(alpha1-2)mannose]mannose (alpha 1-2) mannose (alpha 1-6) mannose (alpha 1-4) glucosamine - (acyl) inositol-phosphate-diacylglycerol (P.ch. alpha) and ethanolamine-phosphate - mannose (alpha 1-2) mannose (alpha 1-6) mannose (alpha 1-4) glucosamine-(acyl)inositol-phosphate-diacylglycerol (P.ch. beta). One of these GPI-anchor precursors (P.ch. alpha) possesses the same carbohydrate structure as the GPI membrane anchor of merozoite surface protein-1 from P.c.chabaudi AS. PMID- 9396738 TI - Effects of adrenaline on triacylglycerol synthesis and turnover in ventricular myocytes from adult rats. AB - Ca2+-tolerant myocytes were isolated with endogenous triacylglycerol (TAG) stores prelabelled with [3H]palmitate and subsequently incubated for a 1h chase period with [14C]palmitate, 2% albumin and 5mM glucose. Measurements were then made of [14C]palmitate conversion into TAG and phospholipids, of loss of [3H]TAG, of glycerol release and of change in the total TAG content. Rates of de novo synthesis of TAG were calculated by a balance method. With 0. 5mM palmitate present, 5 microM adrenaline increased de novo synthesis of TAG by 81% and incorporation of [14C]palmitate into phospholipids by 59%. Significant increases in these processes with adrenaline were also seen with 0.08, 0.14 and 0.26 mM palmitate. The beta-agonist isoprenaline had little effect on de novo synthesis of TAG and had no effect on [14C]palmitate conversion into phospholipids. The alpha1-agonist phenylephrine mimicked adrenaline in increasing [14C]palmitate conversion into phospholipids but had no effect on de novo synthesis of TAG. Adrenaline did not significantly alter the myocyte glycerol 3-phosphate content but caused a persistent 40% increase in the activity of the form of glycerolphosphate acyltransferase found predominantly in the sarcoplasmic reticulum. With 0.5 mM palmitate present, the value [14C]TAG formed -decrease in [3H]TAG consistently exceeded the enzymically measured change in cell TAG content. From this it was suggested that the specific radioactivity of [3H]TAG pool(s) mobilized during the chase period was lower than that of the overall cell TAG. In the basal state, complete mobilization of TAG measured as glycerol release was low, but cycling of TAG to diacylglycerol or monoacylglycerol and back to TAG appeared to be high. With adrenaline present, glycerol release was increased 5-6-fold but recycling of lower acylglycerols to TAG was abolished. Glycerol release was inhibited by increasing extracellular palmitate from 0.08 to 0.5 mM. Adrenaline partially over-rode this effect. PMID- 9396739 TI - Role of membrane-anchored heparin-binding epidermal growth factor-like growth factor and CD9 on macrophages. AB - Heparin-binding epidermal-growth-factor-like growth factor (HB-EGF) is a potent mitogen for smooth-muscle cells (SMCs) belonging to the EGF family. We have previously determined that HB-EGF is expressed in macrophages and SMCs of human atherosclerotic lesions and that its membrane-anchored precursor, proHB-EGF, also has a juxtacrine mitogenic activity which is markedly enhanced by CD9, a surface marker of lymphohaemopoietic cells. Therefore, when both proHB-EGF and CD9 are expressed on macrophages, they may strongly promote the development of atherosclerosis. In the present study we have investigated the changes in proHB EGF and CD9 in THP-1 cells during differentiation into macrophages and by the addition of oxidized low-density lipoproteins (OxLDL) and assessed juxtacrine growth activity of THP-1 macrophages for human aortic SMCs. HB-EGF and CD9 at both the mRNA and the protein level were up-regulated after differentiation into macrophages, and further expression of HB-EGF was induced by the addition of OxLDL or lysophosphatidylcholine. Juxtacrine induction by formalin-fixed growth was suppressed to control levels by an inhibitor of HB-EGF and was partially decreased by anti-CD9 antibodies. These results suggest that co-expression of proHB-EGF and CD9 on macrophages plays an important role in the development of atherosclerosis by a juxtacrine mechanism. PMID- 9396740 TI - Zeta, a novel class of glutathione transferases in a range of species from plants to humans. AB - Sequence alignment and phylogenetic analysis has identified a new subgroup of glutathione S-transferase (GST)-like proteins from a range of species extending from plants to humans. This group has been termed the Zeta class. An atomic model of the N-terminal domain suggests that the members of the Zeta class have a similar structure to that of other GSTs, binding glutathione in a similar orientation in the G site. Recombinant human GSTZ1-1 has been expressed in Escherichia coli and characterized. The protein is a dimer composed of 24.2 kDa subunits and has minimal glutathione-conjugating activity with ethacrynic acid and 7-chloro-4-nitrobenz-2-oxa-1, 3-diazole. Although low in comparison with other GSTs, GSTZ1-1 has glutathione peroxidase activity with t-butyl and cumene hydroperoxides. The members of the Zeta class have been conserved over a long evolutionary period, suggesting that they might have a role in the metabolism of a compound that is common in many living cells. PMID- 9396741 TI - Temperature-sensitive mRNA degradation is an early event in hepatocyte de differentiation. AB - The isolation and culture of metabolically active hepatocytes by proteolytic digestion of the extracellular matrix of the liver results in the transcriptional silencing of liver-specific genes encoding cytochromes P-450 (CYP) and albumin together with an induction of cellular RNase activity. The levels of albumin mRNA are maintained in cultured hepatocytes at similar levels to that present in the intact liver for at least 24 h, whereas the major constitutively expressed CYP2C11 mRNA is rapidly degraded. Hepatocytes heat-shocked at 40 degrees C during the isolation procedure (which results in an induction of heat-shock protein mRNA species) blocks the increase in RNase activity and abrogates the loss of CYP2C11 mRNA for at least 4 h. Cycloheximide-dependent inhibition of protein synthesis blocks the temperature-dependent induction of heat-shock proteins without affecting the protection afforded to CYP2C11 mRNA, indicating that CYP2C11 mRNA levels are not directly dependent on heat-shock protein induction and suggesting that the induction of RNase activity might be responsible for the specific loss of CYP2C11 mRNA in hepatocytes isolated at 37 degrees C. Differential rates of degradation of CYP2C11 transcribed in vitro and of albumin mRNA are observed in the presence of cellular extracts from cultured hepatocytes isolated at 37 degrees C (which have maximally induced levels of cellular RNase activity) but not in comparable extracts from cultured hepatocytes isolated at 40 degrees C, supporting the hypothesis that an RNase activity is induced in culture that specifically degrades CYP2C11 mRNA but not albumin mRNA. These results suggest that an early event in hepatocyte de-differentiation involves the induction of RNase activity in addition to transcriptional silencing of liver-specific genes and that the induced RNase activity demonstrates specificity within liver specific gene products. PMID- 9396742 TI - Serratia marcescens chitobiase is a retaining glycosidase utilizing substrate acetamido group participation. AB - The stereochemistry of the reaction catalysed by Serratia marcescens chitobiase was determined by HPLC separation of the anomers of N-acetylglucosamine produced during the hydrolysis of p-nitrophenyl N-acetyl-beta-d-glucosaminide (PNP GlcNAc). In the early stages of the reaction, the beta-anomer was found to prevail, whereas the alpha-anomer dominated at mutarotation equilibrium. This established that chitobiase hydrolyses glycosidic bonds with overall retention of the anomeric configuration. Chitobiase-catalysed hydrolysis of PNP-GlcNAc was competitively inhibited by a series of chito-oligosaccharides (degree of polymerization 2-5) that were selectively de-N-acetylated at their non-reducing end. The results are in accord with the participation of the acetamido group at C 2 of the substrate in the catalytic mechanism of chitobiase and related enzymes. PMID- 9396743 TI - A chromatin-associated kinesin-related protein required for normal mitotic chromosome segregation in Drosophila. AB - The tiovivo (tio) gene of Drosophila encodes a kinesin-related protein, KLP38B, that colocalizes with condensed chromatin during cell division. Wild-type function of the tio gene product KLP38B is required for normal chromosome segregation during mitosis. Mitotic cells in tio larval brains displayed circular mitotic figures, increased ploidy, and abnormal anaphase figures. KLP38B mRNA is maternally provided and expressed in cells about to undergo division. We propose that KLP38B, perhaps redundantly with other chromosome-associated microtubule motor proteins, contributes to interactions between chromosome arms and microtubules important for establishing bipolar attachment of chromosomes and assembly of stable bipolar spindles. PMID- 9396744 TI - CENP-E function at kinetochores is essential for chromosome alignment. AB - CENP-E is a kinesin-like protein that binds to kinetochores and may provide functions that are critical for normal chromosome motility during mitosis. To directly test the in vivo function of CENP-E, we microinjected affinity-purified antibodies to block the assembly of CENP-E onto kinetochores and then examined the behavior of these chromosomes. Chromosomes lacking CENP-E at their kinetochores consistently exhibited two types of defects that blocked their alignment at the spindle equator. Chromosomes positioned near a pole remained mono-oriented as they were unable to establish bipolar microtubule connections with the opposite pole. Chromosomes within the spindle established bipolar connections that supported oscillations and normal velocities of kinetochore movement between the poles, but these bipolar connections were defective because they failed to align the chromosomes into a metaphase plate. Overexpression of a mutant that lacked the amino-terminal 803 amino acids of CENP-E was found to saturate limiting binding sites on kinetochores and competitively blocked endogenous CENP-E from assembling onto kinetochores. Chromosomes saturated with the truncated CENP-E mutant were never found to be aligned but accumulated at the poles or were strewn within the spindle as was the case when cells were microinjected with CENP-E antibodies. As the motor domain was contained within the portion of CENP-E that was deleted, the chromosomal defect is likely attributed to the loss of motor function. The combined data show that CENP-E provides kinetochore functions that are essential for monopolar chromosomes to establish bipolar connections and for chromosomes with connections to both spindle poles to align at the spindle equator. Both of these events rely on activities that are provided by CENP-E's motor domain. PMID- 9396745 TI - Probing the architecture of a simple kinetochore using DNA-protein crosslinking. AB - In budding yeast, accurate chromosome segregation requires that one and only one kinetochore assemble per chromosome. In this paper, we report the use of DNA protein crosslinking and nondenaturing gel analysis to study the structure of CBF3, a four-protein complex that binds to the essential CDEIII region of Saccharomyces cerevisiae centromeres. We find that three subunits of CBF3 are in direct contact with CDEIII over a region of DNA that spans 80 bp. A highly asymmetric core complex containing p58(CTF13) p64(CEP3) and p110(NDC10) in direct contact with DNA forms at the genetically defined center of CDEIII. This core complex spans approximately 56 bp of CEN3. An extended complex comprising the core complex and additional DNA-bound p110(NDC10) also forms. It spans approximately 80 bp of DNA. CBF3 makes sequence-specific and -nonspecific contacts with DNA. Both contribute significantly to the energy of CBF3-DNA interaction. Moreover, important sequence-specific contacts are made with bases that are not conserved among yeast centromeres. These findings provide a foundation for understanding the organization of the CBF3-centromere complex, a structure that appears to initiate the formation of microtubule attachment sites at yeast kinetochores. These results also have implications for understanding centromere-binding proteins in higher cells. PMID- 9396746 TI - Export of cellubrevin from the endoplasmic reticulum is controlled by BAP31. AB - Cellubrevin is a ubiquitously expressed membrane protein that is localized to endosomes throughout the endocytotic pathway and functions in constitutive exocytosis. We report that cellubrevin binds with high specificity to BAP31, a representative of a highly conserved family of integral membrane proteins that has recently been discovered to be binding proteins of membrane immunoglobulins. The interaction between BAP31 and cellubrevin is sensitive to high ionic strength and appears to require the transmembrane regions of both proteins. No other proteins of liver membrane extracts copurified with BAP31 on immobilized recombinant cellubrevin, demonstrating that the interaction is specific. Synaptobrevin I bound to BAP31 with comparable affinity, whereas only weak binding was detectable with synaptobrevin II. Furthermore, a fraction of BAP31 and cellubrevin was complexed when each of them was quantitatively immunoprecipitated from detergent extracts of fibroblasts (BHK 21 cells). During purification of clathrin-coated vesicles or early endosomes, BAP31 did not cofractionate with cellubrevin. Rather, the protein was enriched in ER-containing fractions. When BHK cells were analyzed by immunocytochemistry, BAP31 did not overlap with cellubrevin, but rather colocalized with resident proteins of the ER. In addition, immunoreactive vesicles were clustered in a paranuclear region close to the microtubule organizing center, but different from the Golgi apparatus. When microtubules were depolymerized with nocodazole, this accumulation disappeared and BAP31 was confined to the ER. Truncation of the cytoplasmic tail of BAP31 prevented export of cellubrevin, but not of the transferrin receptor from the ER. We conclude that BAP31 represents a novel class of sorting proteins that controls anterograde transport of certain membrane proteins from the ER to the Golgi complex. PMID- 9396747 TI - Ceramide accumulation uncovers a cycling pathway for the cis-Golgi network marker, infectious bronchitis virus M protein. AB - The M glycoprotein from the avian coronavirus, infectious bronchitis virus (IBV), contains information for localization to the cis-Golgi network in its first transmembrane domain. We hypothesize that localization to the Golgi complex may depend in part on specific interactions between protein transmembrane domains and membrane lipids. Because the site of sphingolipid synthesis overlaps the localization of IBV M, we asked whether perturbation of sphingolipids affected localization of IBV M. Short-term treatment with two inhibitors of sphingolipid synthesis had no effect on localization of IBV M or other Golgi markers. Thus, ongoing synthesis of these lipids was not required for proper localization. Surprisingly, a third inhibitor, d,l-threo-1-phenyl-2-decanoylamino-3-morpholino- 1-propanol (PDMP), shifted the steady-state distribution of IBV M from the Golgi complex to the ER. This effect was rapid and reversible and was also observed for ERGIC-53 but not for Golgi stack proteins. At the concentration of PDMP used, conversion of ceramide into both glucosylceramide and sphingomyelin was inhibited. Pretreatment with upstream inhibitors partially reversed the effects of PDMP, suggesting that ceramide accumulation mediates the PDMP-induced alterations. Indeed, an increase in cellular ceramide was measured in PDMP treated cells. We propose that IBV M is at least in part localized by retrieval mechanisms. Further, ceramide accumulation reveals this cycle by upsetting the balance of anterograde and retrograde traffic and/ or disrupting retention by altering bilayer dynamics. PMID- 9396748 TI - Peroxisomal targeting, import, and assembly of alcohol oxidase in Pichia pastoris. AB - Alcohol oxidase (AOX), the first enzyme in the yeast methanol utilization pathway is a homooctameric peroxisomal matrix protein. In peroxisome biogenesis-defective (pex) mutants of the yeast Pichia pastoris, AOX fails to assemble into active octamers and instead forms inactive cytoplasmic aggregates. The apparent inability of AOX to assemble in the cytoplasm contrasts with other peroxisomal proteins that are able to oligomerize before import. To further investigate the import of AOX, we first identified its peroxisomal targeting signal (PTS). We found that sequences essential for targeting AOX are primarily located within the four COOH-terminal amino acids of the protein leucine-alanine-arginine phenylalanine COOH (LARF). To examine whether AOX can oligomerize before import, we coexpressed AOX without its PTS along with wild-type AOX and determined whether the mutant AOX could be coimported into peroxisomes. To identify the mutant form of AOX, the COOH-terminal LARF sequence of the protein was replaced with a hemagglutinin epitope tag (AOX-HA). Coexpression of AOX-HA with wild-type AOX (AOX-WT) did not result in an increase in the proportion of AOX-HA present in octameric active AOX, suggesting that newly synthesized AOX-HA cannot oligomerize with AOX-WT in the cytoplasm. Thus, AOX cannot initiate oligomerization in the cytoplasm, but must first be targeted to the organelle before assembly begins. PMID- 9396750 TI - Compartmentalized IgE receptor-mediated signal transduction in living cells. AB - Several receptor-mediated signal transduction pathways, including EGF and IgE receptor pathways, have been proposed to be spatially restricted to plasma membrane microdomains. However, the experimental evidence for signaling events in these microdomains is largely based on biochemical fractionation and immunocytochemical studies and only little is known about their spatial dynamics in living cells. Here we constructed green fluorescent protein-tagged SH2 domains to investigate where and when IgE receptor (FcepsilonRI)-mediated tyrosine phosphorylation occurs in living tumor mast cells. Strikingly, within minutes after antigen addition, tandem SH2 domains from Syk or PLC-gamma1 translocated from a uniform cytosolic distribution to punctuate plasma membrane microdomains. Colocalization experiments showed that the microdomains where tyrosine phosphorylation occurred were indistinguishable from those stained by cholera toxin B, a marker for glycosphingolipids. Competitive binding studies with coelectroporated unlabeled Syk, PLC-gamma1, and other SH2 domains selectively suppressed the induction of IgE receptor-mediated calcium signals as well as the binding of the fluorescent SH2 domains. This supports the hypothesis that PLC gamma1 and Syk SH2 domains selectively bind to Syk and IgE receptors, respectively. Unlike the predicted prelocalization of EGF receptors to caveolae microdomains, fluorescently labeled IgE receptors were found to be uniformly distributed in the plasma membrane of unstimulated cells and only transiently translocated to glycosphingolipid rich microdomains after antigen addition. Thus, these in vivo studies support a plasma membrane signaling mechanism by which IgE receptors transiently associate with microdomains and induce the spatially restricted activation of Syk and PLC-gamma1. PMID- 9396749 TI - Distinct intracellular compartments involved in invariant chain degradation and antigenic peptide loading of major histocompatibility complex (MHC) class II molecules. AB - Major histocompatibility complex (MHC) class II molecules are transported to intracellular MHC class II compartments via a transient association with the invariant chain (Ii). After removal of the invariant chain, peptides can be loaded onto class II molecules, a process catalyzed by human leukocyte antigen-DM (HLA-DM) molecules. Here we show that MHC class II compartments consist of two physically and functionally distinct organelles. Newly synthesized MHC class II/Ii complexes were targeted to endocytic organelles lacking HLA-DM molecules, where Ii degradation occurred. From these organelles, class II molecules were transported to a distinct organelle containing HLA-DM, in which peptides were loaded onto class II molecules. This latter organelle was not directly accessible via fluid phase endocytosis, suggesting that it is not part of the endosomal pathway. Uptake via antigen-specific membrane immunoglobulin resulted however in small amounts of antigen in the HLA-DM positive organelles. From this peptide loading compartment, class II-peptide complexes were transported to the plasma membrane, in part after transit through endocytic organelles. The existence of two separate compartments, one involved in Ii removal and the other functioning in HLA-DM-dependent peptide loading of class II molecules, may contribute to the efficiency of antigen presentation by the selective recruitment of peptide receptive MHC class II molecules and HLA-DM to the same subcellular location. PMID- 9396751 TI - Activation of a retroviral membrane fusion protein: soluble receptor-induced liposome binding of the ALSV envelope glycoprotein. AB - It is not known how membrane fusion proteins that function at neutral pH, for example the human immunodeficiency virus envelope (Env) glycoprotein and intracellular fusion machines, are activated for target bilayer binding. We have addressed this question using a soluble oligomeric form of an avian retroviral Env glycoprotein (API) and soluble forms of its receptor. Binding of soluble receptor to API induces API to bind to liposomes composed of phosphatidylcholine and cholesterol at neutral pH. Liposome binding only occurs at fusion permissive temperatures (T > 20 degrees C), is complete between 2 to 5 min at 37 degrees C, and is stable to high salt, carbonate, and urea. Liposome binding is mediated by the ectodomain of the transmembrane subunit of API, and a mutant with a Val to Glu substitution in the Env fusion peptide (located in the ectodomain of the transmembrane subunit) shows significantly reduced liposome binding. Moreover, under conditions of equivalent binding to API, a mutant receptor that does not support infection (Zingler, K., and J.A.T. Young. 1996. J. Virol. 70:7510-7516) does not induce significant liposome binding. Our results indicate that a highly specific interaction between an avian retroviral Env and its receptor activates the retroviral glycoprotein for target bilayer binding at neutral pH in much the same way as low pH activates the influenza hemagglutinin. Our findings are discussed in terms of the mechanisms of viral and cellular fusion proteins that function at neutral pH. PMID- 9396752 TI - Direct visualization of the translocation of the gamma-subspecies of protein kinase C in living cells using fusion proteins with green fluorescent protein. AB - We expressed the gamma-subspecies of protein kinase C (gamma-PKC) fused with green fluorescent protein (GFP) in various cell lines and observed the movement of this fusion protein in living cells under a confocal laser scanning fluorescent microscope. gamma-PKC-GFP fusion protein had enzymological properties very similar to that of native gamma-PKC. The fluorescence of gamma-PKC- GFP was observed throughout the cytoplasm in transiently transfected COS-7 cells. Stimulation by an active phorbol ester (12-O-tetradecanoylphorbol 13-acetate [TPA]) but not by an inactive phorbol ester (4alpha-phorbol 12, 13-didecanoate) induced a significant translocation of gamma-PKC-GFP from cytoplasm to the plasma membrane. A23187, a Ca2+ ionophore, induced a more rapid translocation of gamma PKC-GFP than TPA. The A23187-induced translocation was abolished by elimination of extracellular and intracellular Ca2+. TPA- induced translocation of gamma-PKC GFP was unidirected, while Ca2+ ionophore-induced translocation was reversible; that is, gamma-PKC-GFP translocated to the membrane returned to the cytosol and finally accumulated as patchy dots on the plasma membrane. To investigate the significance of C1 and C2 domains of gamma-PKC in translocation, we expressed mutant gamma-PKC-GFP fusion protein in which the two cysteine rich regions in the C1 region were disrupted (designated as BS 238) or the C2 region was deleted (BS 239). BS 238 mutant was translocated by Ca2+ ionophore but not by TPA. In contrast, BS 239 mutant was translocated by TPA but not by Ca2+ ionophore. To examine the translocation of gamma-PKC-GFP under physiological conditions, we expressed it in NG-108 cells, N-methyl-D-aspartate (NMDA) receptor-transfected COS-7 cells, or CHO cells expressing metabotropic glutamate receptor 1 (CHO/mGluR1 cells). In NG-108 cells , K+ depolarization induced rapid translocation of gamma-PKC-GFP. In NMDA receptor-transfected COS-7 cells, application of NMDA plus glycine also translocated gamma-PKC-GFP. Furthermore, rapid translocation and sequential retranslocation of gamma-PKC-GFP were observed in CHO/ mGluR1 cells on stimulation with the receptor. Neither cytochalasin D nor colchicine affected the translocation of gamma-PKC-GFP, indicating that translocation of gamma-PKC was independent of actin and microtubule. gamma-PKC GFP fusion protein is a useful tool for investigating the molecular mechanism of gamma-PKC translocation and the role of gamma-PKC in the central nervous system. PMID- 9396754 TI - The Arabidopsis KNOLLE protein is a cytokinesis-specific syntaxin. AB - In higher plant cytokinesis, plasma membrane and cell wall originate by vesicle fusion in the plane of cell division. The Arabidopsis KNOLLE gene, which is required for cytokinesis, encodes a protein related to vesicle-docking syntaxins. We have raised specific rabbit antiserum against purified recombinant KNOLLE protein to show biochemically and by immunoelectron microscopy that KNOLLE protein is membrane associated. Using immunofluorescence microscopy, KNOLLE protein was found to be specifically expressed during mitosis and, unlike the plasma membrane H+-ATPase, to localize to the plane of division during cytokinesis. Arabidopsis dynamin-like protein ADL1 accumulates at the plane of cell plate formation in knolle mutant cells as in wild-type cells, suggesting that cytokinetic vesicle traffic is not affected. Furthermore, electron microscopic analysis indicates that vesicle fusion is impaired. KNOLLE protein was detected in mitotically dividing cells of various parts of the developing plant, including seedling root, inflorescence meristem, floral meristems and ovules, and the cellularizing endosperm, but not during cytokinesis after the male second meiotic division. Thus, KNOLLE is the first syntaxin-like protein that appears to be involved specifically in cytokinetic vesicle fusion. PMID- 9396753 TI - Sarcomeric gene expression and contractility in myofibroblasts. AB - Myofibroblasts are unusual cells that share morphological and functional features of muscle and nonmuscle cells. Such cells are thought to control liver blood flow and kidney glomerular filtration rate by having unique contractile properties. To determine how these cells achieve their contractile properties and their resemblance to muscle cells, we have characterized two myofibroblast cell lines. Here, we demonstrate that myofibroblast cell lines from kidney mesangial cells (BHK) and liver stellate cells activate extensive programs of muscle gene expression including a wide variety of muscle structural proteins. In BHK cells, six different striated myosin heavy chain isoforms and many thin filament proteins, including troponin T and tropomyosin are expressed. Liver stellate cells express a limited subset of the muscle thick filament proteins expressed in BHK cells. Although these cells are mitotically active and do not morphologically differentiate into myotubes, we show that MyoD and myogenin are expressed and functional in both cell types. Finally, these cells contract in response to endothelin-1 (ET-1); and we show that ET-1 treatment increases the expression of sarcomeric myosin. PMID- 9396755 TI - The axonal membrane protein Caspr, a homologue of neurexin IV, is a component of the septate-like paranodal junctions that assemble during myelination. AB - We have investigated the potential role of contactin and contactin-associated protein (Caspr) in the axonal-glial interactions of myelination. In the nervous system, contactin is expressed by neurons, oligodendrocytes, and their progenitors, but not by Schwann cells. Expression of Caspr, a homologue of Neurexin IV, is restricted to neurons. Both contactin and Caspr are uniformly expressed at high levels on the surface of unensheathed neurites and are downregulated during myelination in vitro and in vivo. Contactin is downregulated along the entire myelinated nerve fiber. In contrast, Caspr expression initially remains elevated along segments of neurites associated with nascent myelin sheaths. With further maturation, Caspr is downregulated in the internode and becomes strikingly concentrated in the paranodal regions of the axon, suggesting that it redistributes from the internode to these sites. Caspr expression is similarly restricted to the paranodes of mature myelinated axons in the peripheral and central nervous systems; it is more diffusely and persistently expressed in gray matter and on unmyelinated axons. Immunoelectron microscopy demonstrated that Caspr is localized to the septate-like junctions that form between axons and the paranodal loops of myelinating cells. Caspr is poorly extracted by nonionic detergents, suggesting that it is associated with the axon cytoskeleton at these junctions. These results indicate that contactin and Caspr function independently during myelination and that their expression is regulated by glial ensheathment. They strongly implicate Caspr as a major transmembrane component of the paranodal junctions, whose molecular composition has previously been unknown, and suggest its role in the reciprocal signaling between axons and glia. PMID- 9396756 TI - Distribution and function of laminins in the neuromuscular system of developing, adult, and mutant mice. AB - Laminins, heterotrimers of alpha, beta, and gamma chains, are prominent constituents of basal laminae (BLs) throughout the body. Previous studies have shown that laminins affect both myogenesis and synaptogenesis in skeletal muscle. Here we have studied the distribution of the 10 known laminin chains in muscle and peripheral nerve, and assayed the ability of several heterotrimers to affect the outgrowth of motor axons. We show that cultured muscle cells express four different alpha chains (alpha1, alpha2, alpha4, and alpha5), and that developing muscles incorporate all four into BLs. The portion of the muscle's BL that occupies the synaptic cleft contains at least three alpha chains and two beta chains, but each is regulated differently. Initially, the alpha2, alpha4, alpha5, and beta1 chains are present both extrasynaptically and synaptically, whereas beta2 is restricted to synaptic BL from its first appearance. As development proceeds, alpha2 remains broadly distributed, whereas alpha4 and alpha5 are lost from extrasynaptic BL and beta1 from synaptic BL. In adults, alpha4 is restricted to primary synaptic clefts whereas alpha5 is present in both primary and secondary clefts. Thus, adult extrasynaptic BL is rich in laminin 2 (alpha2beta1gamma1), and synaptic BL contains laminins 4 (alpha2beta2gamma1), 9 (alpha4beta2gamma1), and 11 (alpha5beta2gamma1). Likewise, in cultured muscle cells, alpha2 and beta1 are broadly distributed but alpha5 and beta2 are concentrated at acetylcholine receptor-rich "hot spots," even in the absence of nerves. The endoneurial and perineurial BLs of peripheral nerve also contain distinct laminin chains: alpha2, beta1, gamma1, and alpha4, alpha5, beta2, gamma1, respectively. Mutation of the laminin alpha2 or beta2 genes in mice not only leads to loss of the respective chains in both nerve and muscle, but also to coordinate loss and compensatory upregulation of other chains. Notably, loss of beta2 from synaptic BL in beta2(-/-) "knockout" mice is accompanied by loss of alpha5, and decreased levels of alpha2 in dystrophic alpha2(dy/dy) mice are accompanied by compensatory retention of alpha4. Finally, we show that motor axons respond in distinct ways to different laminin heterotrimers: they grow freely between laminin 1 (alpha1beta1gamma1) and laminin 2, fail to cross from laminin 4 to laminin 1, and stop upon contacting laminin 11. The ability of laminin 11 to serve as a stop signal for growing axons explains, in part, axonal behaviors observed at developing and regenerating synapses in vivo. PMID- 9396757 TI - Ligation of major histocompatability complex (MHC) class I molecules on human T cells induces cell death through PI-3 kinase-induced c-Jun NH2-terminal kinase activity: a novel apoptotic pathway distinct from Fas-induced apoptosis. AB - Ligation of major histocompatability complex class I (MHC-I) molecules expressed on T cells leads to both growth arrest and apoptosis. The aim of the current study was to investigate the intracellular signal pathways that mediate these effects. MHC-I ligation of human Jurkat T cells induced a morphologically distinct form of apoptosis within 6 h. A specific caspase inhibitor, which inhibited Fas-induced apoptosis, did not affect apoptosis induced by MHC-I ligation. Furthermore, MHC-I-induced apoptosis did not involve cleavage and activation of the poly(ADP- ribose) polymerase (PARP) endonuclease or degradation of genomic DNA into the typical fragmentation ladder, both prominent events of Fas-induced apoptosis. These results suggest that MHC-I ligation of Jurkat T cells induce apoptosis through a signal pathway distinct from the Fas molecule. In our search for other signal pathways leading to apoptosis, we found that the regulatory 85-kD subunit of the phosphoinositide-3 kinase (PI-3) kinase was tyrosine phosphorylated after ligation of MHC-I and the PI-3 kinase inhibitor wortmannin selectively blocked MHC-I-, but not Fas-induced, apoptosis. As the c Jun NH2-terminal kinase (JNK) can be activated by PI-3 kinase activity, and has been shown to be involved in apoptosis of lymphocytes, we examined JNK activation after MHC-I ligation. Strong JNK activity was observed after MHC-I ligation and the activity was completely blocked by wortmannin. Inhibition of JNK activity, by transfecting cells with a dominant-negative JNKK- MKK4 construct, led to a strong reduction of apoptosis after MHC-I ligation. These results suggest a critical engagement of PI-3 kinase-induced JNK activity in apoptosis induced by MHC-I ligation. PMID- 9396758 TI - Thyroid hormone induces apoptosis in primary cell cultures of tadpole intestine: cell type specificity and effects of extracellular matrix. AB - Thyroid hormone (T3 or 3,5,3'-triiodothyronine) plays a causative role during amphibian metamorphosis. To investigate how T3 induces some cells to die and others to proliferate and differentiate during this process, we have chosen the model system of intestinal remodeling, which involves apoptotic degeneration of larval epithelial cells and proliferation and differentiation of other cells, such as the fibroblasts and adult epithelial cells, to form the adult intestine. We have established in vitro culture conditions for intestinal epithelial cells and fibroblasts. With this system, we show that T3 can enhance the proliferation of both cell types. However, T3 also concurrently induces larval epithelial apoptosis, which can be inhibited by the extracellular matrix (ECM). Our studies with known inhibitors of mammalian cell death reveal both similarities and differences between amphibian and mammalian cell death. These, together with gene expression analysis, reveal that T3 appears to simultaneously induce different pathways that lead to specific gene regulation, proliferation, and apoptotic degeneration of the epithelial cells. Thus, our data provide an important molecular and cellular basis for the differential responses of different cell types to the endogenous T3 during metamorphosis and support a role of ECM during frog metamorphosis. PMID- 9396759 TI - Embryonic expression of the putative gamma subunit of the sodium pump is required for acquisition of fluid transport capacity during mouse blastocyst development. AB - The sodium/potassium pump, Na+,K+-ATPase, is generally understood to function as a heterodimer of two subunits, a catalytic alpha subunit and a noncatalytic, glycosylated beta subunit. Recently, a putative third subunit, the gamma subunit, was cloned. This small protein (6.5 kD) coimmunoprecipitates with the alpha and beta subunits and is closely associated with the ouabain binding site on the holoenzyme, but its function is unknown. We have investigated the expression of the gamma subunit in preimplantation mouse development, where Na+, K+-ATPase plays a critical role as the driving force for blastocoel formation (cavitation). Using reverse transcriptase-polymerase chain reaction, we demonstrated that the gamma subunit mRNA accumulates continuously from the eight-cell stage onward and that it cosediments with polyribosomes from its time of first appearance. Confocal immunofluorescence microscopy revealed that the gamma subunit itself accumulates and is localized at the blastomere surfaces up to the blastocyst stage. In contrast with the alpha and beta subunits, the gamma subunit is not concentrated in the basolateral surface of the polarized trophectoderm layer, but is strongly expressed at the apical surface as well. When embryos were treated with antisense oligodeoxynucleotide complementary to the gamma subunit mRNA, ouabain-sensitive K+ transport (as indicated by 86Rb+ uptake) was reduced and cavitation delayed. However, Na+, K+-ATPase enzymatic activity was unaffected as determined by a direct phosphorylation assay ("back door" phosphorylation) applied to plasma membrane preparations. These results indicate that the gamma subunit, although not an integral component of Na+,K+-ATPase, is an important determinant of active cation transport and that, as such, its embryonic expression is essential for blastocoel formation in the mouse. PMID- 9396760 TI - Key role of the Cdx2 homeobox gene in extracellular matrix-mediated intestinal cell differentiation. AB - To explore the role of homeobox genes in the intestine, the human colon adenocarcinoma cell line Caco2-TC7 has been stably transfected with plasmids synthesizing Cdx1 and Cdx2 sense and antisense RNAs. Cdx1 overexpression or inhibition by antisense RNA does not markedly modify the cell differentiation markers analyzed in this study. In contrast, Cdx2 overexpression stimulates two typical markers of enterocytic differentiation: sucrase-isomaltase and lactase. Cells in which the endogenous expression of Cdx2 is reduced by antisense RNA attach poorly to the substratum. Conversely, Cdx2 overexpression modifies the expression of molecules involved in cell-cell and cell-substratum interactions and in transduction process: indeed, E-cadherin, integrin-beta4 subunit, laminin gamma2 chain, hemidesmosomal protein, APC, and alpha-actinin are upregulated. Interestingly, most of these molecules are preferentially expressed in vivo in the differentiated villi enterocytes rather than in crypt cells. Cdx2 overexpression also results in the stimulation of HoxA-9 mRNA expression, an homeobox gene selectively expressed in the colon. In contrast, Cdx2 overexpressing cells display a decline of Cdx1 mRNA, which is mostly found in vivo in crypt cells. When implanted in nude mice, Cdx2-overexpressing cells produce larger tumors than control cells, and form glandular and villus-like structures. Laminin-1 is known to stimulate intestinal cell differentiation in vitro. In the present study, we demonstrate that the differentiating effect of laminin-1 coatings on Caco2-TC7 cells is accompanied by an upregulation of Cdx2. To further document this observation, we analyzed a series of Caco2 clones in which the production of laminin-alpha1 chain is differentially inhibited by antisense RNA. We found a positive correlation between the level of Cdx2 expression, that of endogenous laminin-alpha1 chain mRNA and that of sucrase isomaltase expression in these cell lines. Taken together, these results suggest (a) that Cdx1 and Cdx2 homeobox genes play distinct roles in the intestinal epithelium, (b) that Cdx2 provokes pleiotropic effects triggering cells towards the phenotype of differentiated villus enterocytes, and (c) that Cdx2 expression is modulated by basement membrane components. Hence, we conclude that Cdx2 plays a key role in the extracellular matrix-mediated intestinal cell differentiation. PMID- 9396761 TI - A single immunoglobulin-like domain of the human neural cell adhesion molecule L1 supports adhesion by multiple vascular and platelet integrins. AB - The neural cell adhesion molecule L1 has been shown to function as a homophilic ligand in a variety of dynamic neurological processes. Here we demonstrate that the sixth immunoglobulin-like domain of human L1 (L1-Ig6) can function as a heterophilic ligand for multiple members of the integrin superfamily including alphavbeta3, alphavbeta1, alpha5beta1, and alphaIIbbeta3. The interaction between L1-Ig6 and alphaIIbbeta3 was found to support the rapid attachment of activated human platelets, whereas a corresponding interaction with alphavbeta3 and alphavbeta1 supported the adhesion of umbilical vein endothelial cells. Mutation of the single Arg-Gly-Asp (RGD) motif in human L1-Ig6 effectively abrogated binding by the aforementioned integrins. A L1 peptide containing this RGD motif and corresponding flanking amino acids (PSITWRGDGRDLQEL) effectively blocked L1 integrin interactions and, as an immobilized ligand, supported adhesion via alphavbeta3, alphavbeta1, alpha5beta1, and alphaIIbbeta3. Whereas beta3 integrin binding to L1-Ig6 was evident in the presence of either Ca2+, Mg2+, or Mn2+, a corresponding interaction with the beta1 integrins was only observed in the presence of Mn2+. Furthermore, such Mn2+-dependent binding by alpha5beta1 and alphavbeta1 was significantly inhibited by exogenous Ca2+. Our findings suggest that physiological levels of calcium will impose a hierarchy of integrin binding to L1 such that alphavbeta3 or active alphaIIbbeta3 > alphavbeta1 > alpha5beta1. Given that L1 can interact with multiple vascular or platelet integrins it is significant that we also present evidence for de novo L1 expression on blood vessels associated with certain neoplastic or inflammatory diseases. Together these findings suggest an expanded and novel role for L1 in vascular and thrombogenic processes. PMID- 9396762 TI - Muscle beta1D integrin reinforces the cytoskeleton-matrix link: modulation of integrin adhesive function by alternative splicing. AB - Expression of muscle-specific beta1D integrin with an alternatively spliced cytoplasmic domain in CHO and GD25, beta1 integrin-minus cells leads to their phenotypic conversion. beta1D-transfected nonmuscle cells display rounded morphology, lack of pseudopodial activity, retarded spreading, reduced migration, and significantly enhanced contractility compared with their beta1A-expressing counterparts. The transfected beta1D is targeted to focal adhesions and efficiently displaces the endogenous beta1A and alphavbeta3 integrins from the sites of cell-matrix contact. This displacement is observed on several types of extracellular matrix substrata and leads to elevated stability of focal adhesions in beta1D transfectants. Whereas a significant part of cellular beta1A integrin is extractable in digitonin, the majority of the transfected beta1D is digitonin insoluble and is strongly associated with the detergent-insoluble cytoskeleton. Increased interaction of beta1D integrin with the actin cytoskeleton is consistent with and might be mediated by its enhanced binding to talin. In contrast, beta1A interacts more strongly with alpha-actinin, than beta1D. Inside out driven activation of the beta1D ectodomain increases ligand binding and fibronectin matrix assembly by beta1D transfectants. Phenotypic effects of beta1D integrin expression in nonmuscle cells are due to its enhanced interactions with both cytoskeletal and extracellular ligands. They parallel the transitions that muscle cells undergo during differentiation. Modulation of beta1 integrin adhesive function by alternative splicing serves as a physiological mechanism reinforcing the cytoskeleton- matrix link in muscle cells. This reflects the major role for beta1D integrin in muscle, where extremely stable association is required for contraction. PMID- 9396763 TI - The thrombopoietin receptor can mediate proliferation without activation of the Jak-STAT pathway. AB - Cytokine receptors of the hematopoietic receptor superfamily lack intrinsic tyrosine kinase domains for the intracellular transmission of their signals. Instead all members of this family associate with Jak family nonreceptor tyrosine kinases. Upon ligand stimulation of the receptors, Jaks are activated to phosphorylate target substrates. These include STAT (signal transducers and activators of transcription) proteins, which after phosphorylation translocate to the nucleus and modulate gene expression. The exact role of the Jak-STAT pathway in conveying growth and differentiation signals remains unclear. Here we describe a deletion mutant of the thrombopoietin receptor (c-mpl) that has completely lost the capacity to activate Jaks and STATs but retains its ability to induce proliferation. This mutant still mediates TPO-induced phosphorylation of Shc, Vav, mitogen-activated protein kinase (MAPK) and Raf-1 as well as induction of c fos and c-myc, although at somewhat reduced levels. Furthermore, we show that both wild-type and mutant receptors activate phosphatidylinositol (PI) 3-kinase upon thrombopoietin stimulation and that thrombopoietin-induced proliferation is inhibited in the presence of the PI 3-kinase inhibitor wortmannin. These results demonstrate that the Jak-STAT pathway is dispensable for the generation of mitogenic signals by a cytokine receptor. PMID- 9396764 TI - Association with FcRgamma is essential for activation signal through NKR-P1 (CD161) in natural killer (NK) cells and NK1.1+ T cells. AB - Natural killer (NK) cells exhibit cytotoxicity against variety of tumor cells and virus-infected cells without prior sensitization and represent unique lymphocytes involved in primary host defense. NKR-P1 is thought to be one of NK receptors mediating activation signals because cross-linking of NKR-P1 activates NK cells to exhibit cytotoxicity and IFN-gamma production. However, molecular mechanism of NK cell activation via NKR-P1 is not well elucidated. In this study, we analyzed the cell surface complex associated with NKR-P1 on NK cells and found that NKR-P1 associates with the FcRgamma chain which is an essential component of Fc receptors for IgG and IgE. The association between FcRgamma and NKR-P1 is independent of Fc receptor complexes. Furthermore, NK cells from FcRgamma deficient mice did not show cytotoxicity or IFN-gamma production upon NKR-P1 cross-linking. Similarly, NK1.1+ T cells from FcRgamma-deficient mice did not produce IFN-gamma upon NKR-P1 crosslinking. These findings demonstrate that the FcRgamma chain plays an important role in activation of NK cells via the NKR-P1 molecule. PMID- 9396765 TI - Functional role for Syk tyrosine kinase in natural killer cell-mediated natural cytotoxicity. AB - Natural killer (NK) cells are named based on their natural cytotoxic activity against a variety of target cells. However, the mechanisms by which sensitive targets activate killing have been difficult to study due to the lack of a prototypic NK cell triggering receptor. Pharmacologic evidence has implicated protein tyrosine kinases (PTKs) in natural killing; however, Lck-deficient, Fyn deficient, and ZAP-70-deficient mice do not exhibit defects in natural killing despite demonstrable defects in T cell function. This discrepancy implies the involvement of other tyrosine kinases. Here, using combined biochemical, pharmacologic, and genetic approaches, we demonstrate a central role for the PTK Syk in natural cytotoxicity. Biochemical analyses indicate that Syk is tyrosine phosphorylated after stimulation with a panel of NK-sensitive target cells. Pharmacologic exposure to piceatannol, a known Syk family kinase inhibitor, inhibits natural cytotoxicity. In addition, gene transfer of dominant-negative forms of Syk to NK cells inhibits natural cytotoxicity. Furthermore, sensitive targets that are rendered NK-resistant by major histocompatibility complex (MHC) class I transfection no longer activate Syk. These data suggest that Syk activation is an early and requisite signaling event in the development of natural cytotoxicity directed against a variety of cellular targets. PMID- 9396766 TI - The E1B 19K/Bcl-2-binding protein Nip3 is a dimeric mitochondrial protein that activates apoptosis. AB - Nip3 (nineteen kD interacting protein-3) is an E1B 19K and Bcl-2 binding protein of unknown function. Nip3 is detected as both a 60- and 30-kD protein in vivo and in vitro and exhibits strong homologous interaction in a yeast two-hybrid system indicating that it can homodimerize. Nip3 is expressed in mitochondria and a mutant (Nip3(163)) lacking the putative transmembrane domain and COOH terminus does not dimerize or localize to mitochondria. Transient transfection of epitope tagged Nip3 in Rat-1 fibroblasts and MCF-7 breast carcinoma induces apoptosis within 12 h while cells transfected with the Nip3(163) mutant have a normal phenotype, suggesting that mitochondrial localization is necessary for induction of cell death. Nip3 overexpression increases the sensitivity to apoptosis induced by granzyme B and topoisomerase I and II inhibitors. After transfection, both Nip3 and Nip3(163) protein levels decrease steadily over 48 h indicating that the protein is rapidly degraded and this occurs in the absence of cell death. Bcl-2 overexpression initially delays the onset of apoptosis induced by Nip3 but the resistance is completely overcome in longer periods of incubation. Nip3 protein levels are much higher and persist longer in Bcl-2 expressing cells. In conclusion, Nip3 is an apoptosis-inducing dimeric mitochondrial protein that can overcome Bcl-2 suppression. PMID- 9396767 TI - Induction of apoptosis of metastatic mammary carcinoma cells in vivo by disruption of tumor cell surface CD44 function. AB - To understand how the hyaluronan receptor CD44 regulates tumor metastasis, the murine mammary carcinoma TA3/St, which constitutively expresses cell surface CD44, was transfected with cDNAs encoding soluble isoforms of CD44 and the transfectants (TA3sCD44) were compared with parental cells (transfected with expression vector only) for growth in vivo and in vitro. Local release of soluble CD44 by the transfectants inhibited the ability of endogenous cell surface CD44 to bind and internalize hyaluronan and to mediate TA3 cell invasion of hyaluronan producing cell monolayers. Mice intravenously injected with parental TA3/St cells developed massive pulmonary metastases within 21-28 d, whereas animals injected with TA3sCD44 cells developed few or no tumors. Tracing of labeled parental and transfectant tumor cells revealed that both cell types initially adhered to pulmonary endothelium and penetrated the interstitial stroma. However, although parental cells were dividing and forming clusters within lung tissue 48 h following injection, >80% of TA3sCD44 cells underwent apoptosis. Although sCD44 transfectants displayed a marked reduction in their ability to internalize and degrade hyaluronan, they elicited abundant local hyaluronan production within invaded lung tissue, comparable to that induced by parental cells. These observations provide direct evidence that cell surface CD44 function promotes tumor cell survival in invaded tissue and that its suppression can induce apoptosis of the invading tumor cells, possibly as a result of impairing their ability to penetrate the host tissue hyaluronan barrier. PMID- 9396768 TI - Distinct roles of lymphotoxin alpha and the type I tumor necrosis factor (TNF) receptor in the establishment of follicular dendritic cells from non-bone marrow derived cells. AB - In mice deficient in either lymphotoxin alpha (LT-alpha) or type I tumor necrosis factor receptor (TNFR-I), organized clusters of follicular dendritic cells (FDC) and germinal centers (GC) are absent from the spleen. We investigated the role of LT-alpha and TNFR-I in the establishment of spleen FDC and GC structure by using reciprocal bone marrow (BM) transfer. When LT-alpha-deficient mice were reconstituted with wild-type BM, FDC organization and the ability to form GC were restored, indicating that the LT-alpha-expressing cells required to establish organized FDC are derived from BM. The role of LT-alpha in establishing organized FDC structure was further investigated by the transfer of complement receptor 1 and 2 (CR1/2)-deficient BM cells into LT-alpha-deficient mice. Organized FDC were identified with both the FDC-M1 and anti-CR1 monoclonal antibodies in these BM chimeric mice, indicating that these cells were derived from the LT-alpha deficient recipient. Thus, expression of LT-alpha in the BM-derived cells, but not in the non-BM-derived cells, is required for the maturation of FDC from non BM precursor cells. In contrast, when TNFR-I-deficient mice were reconstituted with wild-type BM, they showed no detectable FDC clusters or GC formation. This indicates that TNFR-I expression on non-BM-derived cellular components is necessary for the establishment of these lymphoid structures. TNFR-I-deficient BM was able to restore FDC organization and GC formation in LT-alpha-deficient mice, indicating that formation of these structures does not require TNFR-I expression on BM-derived cells. The data in this study demonstrate that FDC organization and GC formation are controlled by both LT-alpha-expressing BM-derived cells and by TNFR-I-expressing non-BM-derived cells. PMID- 9396769 TI - Self-reactive B cells are not eliminated or inactivated by autoantigen expressed on thyroid epithelial cells. AB - Graves' Disease results from the production of autoantibodies against receptors for thyroid stimulating hormone (TSH) on thyroid epithelial cells, and represents the prototype for numerous autoimmune diseases caused by autoantibodies that bind to organ-specific cell membrane antigens. To study how humoral tolerance is normally maintained to organ-specific membrane antigens, transgenic mice were generated selectively expressing membrane-bound hen egg lysozyme (mHEL) on the thyroid epithelium. In contrast to the deletion of autoreactive B cells triggered by systemic mHEL (Hartley, S.B., J. Crosbie, R. Brink, A.B. Kantor, A. Basten, and C.C. Goodnow. 1991. Nature. 353:765-769), selective expression of mHEL autoantigen on thyroid cells did not trigger elimination or inactivation of circulating HEL-reactive B cells. These results provide evidence that tolerance is not actively acquired to organ-specific antigens in the preimmune B cell repertoire, underscoring the importance of maintaining tolerance to such antigens by other mechanisms. The role of an intact endothelial barrier in sequestering organ-specific antigens from circulating preimmune B cells is discussed. PMID- 9396771 TI - Interferon gamma gene expression in sensory neurons: evidence for autocrine gene regulation. AB - We explored expression and possible function of interferon-gamma (IFN-gamma) in cultured fetal (E15) rat dorsal root ganglion neurons combining whole cell patch clamp electrophysiology with single cell reverse transcriptase polymerase chain reaction and confocal laser immunocytochemistry. Morphologically, we located IFN gamma protein in the cytoplasm of the neurons in culture as well as in situ during peri- and postnatal development. Transcripts for classic IFN-gamma and for its receptor were determined in probes of cytoplasm sampled from individual cultured neurons, which had been identified by patch clamp electrophysiology. In addition, the cultured neurons expressed both chains of the IFN-gamma receptor. Locally produced IFN-gamma acts back on its cellular source. Phosphorylation and nuclear translocation of the IFN-inducible transcriptional factor STAT1 as well as IFN-gamma-dependent expression of major histocompatibility complex class I molecules on the neuronal membrane were noted in untreated cultures. However, both processes were substantially blocked in the presence of antibodies neutralizing IFN-gamma. Our findings indicate a role of IFN-gamma in autocrine regulation of sensory neurons. PMID- 9396770 TI - Syk tyrosine kinase is required for the positive selection of immature B cells into the recirculating B cell pool. AB - The tyrosine kinase Syk has been implicated as a key signal transducer from the B cell antigen receptor (BCR). We show here that mutation of the Syk gene completely blocks the maturation of immature B cells into recirculating cells and stops their entry into B cell follicles. Furthermore, using radiation chimeras we demonstrate that this developmental block is due to the absence of Syk in the B cells themselves. Syk-deficient B cells are shown to have the life span of normal immature B cells. If this is extended by over-expression of Bcl-2, they accumulate in the T zone and red pulp of the spleen in increased numbers, but still fail to mature to become recirculating follicular B cells. Despite this defect in maturation, Syk-deficient B cells were seen to give rise to switched as well as nonswitched splenic plasma cells. Normally only a proportion of immature B cells is recruited into the recirculating pool. Our results suggest that Syk transduces a BCR signal that is absolutely required for the positive selection of immature B cells into the recirculating B cell pool. PMID- 9396772 TI - Role of CD8 in aberrant function of cytotoxic T lymphocytes. AB - Using H-2Kd-restricted photoprobe-specific cytotoxic T lymphocyte (CTL) clones, which permit assessment of T cell receptor (TCR)-ligand interactions by TCR photoaffinity labeling, we observed that the efficiency of antigen recognition by CTL was critically dependent on the half-life of TCR-ligand complexes. We show here that antigen recognition by CTL is essentially determined by the frequency of serial TCR engagement, except for very rapid dissociations, which resulted in aberrant TCR signaling and antagonism. Thus agonists that were efficiently recognized exhibited rapid TCR-ligand complex dissociation, and hence a high frequency of serial TCR engagement, whereas the opposite was true for weak agonists. Surprisingly, these differences were largely accounted for by the coreceptor CD8. While it was known that CD8 substantially decreases TCR-ligand complex dissociation, we observed in this study that this effect varied considerably among ligand variants, indicating that epitope modifications can alter the CD8 contribution to TCR-ligand binding, and hence the efficiency of antigen recognition by CTL. PMID- 9396773 TI - Determinant spreading of T helper cell 2 (Th2) responses to pancreatic islet autoantigens. AB - The nature (Th1 versus Th2) and dynamics of the autoimmune response during the development of insulin-dependent diabetes mellitus (IDDM) and after immunotherapy are unclear. Here, we show in nonobese diabetic (NOD) mice that the autoreactive T cell response starts and spreads as a pure Th1 type autoimmunity, suggesting that a spontaneous Th1 cascade underlies disease progression. Surprisingly, induction of antiinflammatory Th2 responses to a single beta cell antigen (betaCA) resulted in the spreading of Th2 cellular and humoral immunity to unrelated betaCAs in an infectious manner and protection from IDDM. The data suggest that both Th1 and Th2 autoimmunity evolve in amplificatory cascades by generating site-specific, but not antigen-specific, positive feedback circuits. Determinant spreading of Th2 responses may be a fundamental mechanism underlying antigen-based immunotherapeutics, explaining observations of infectious tolerance and providing a new theoretical framework for therapeutic intervention. PMID- 9396774 TI - Membrane Fas ligand kills human peripheral blood T lymphocytes, and soluble Fas ligand blocks the killing. AB - It has been believed that the Fas expressed on human peripheral blood T cells (PBT) is nonfunctional, because these cells are insensitive to agonistic anti Fas/Apo-1 mAbs that efficiently kill in vitro-activated T cells and many Fas expressing cell lines. Here, we demonstrate that membrane-bound Fas ligand (FasL) kills both fresh and in vitro-activated PBT, indicating that the Fas expressed on fresh PBT is functional. In contrast, soluble FasL kills only the latter. Naive T cells in umbilical cord blood do not express Fas, but can be induced to express Fas by IFN-gamma or by a combination of IL-2 and anti-CD28 mAb, after which they acquire sensitivity to membrane but not to soluble FasL. Soluble FasL inhibited the killing of fresh PBT by membrane FasL. These results indicate that the shedding of FasL from the membrane is a mechanism for downregulating at least part of its killing activity. PMID- 9396775 TI - Targeted deletion of the lipopolysaccharide (LPS)-binding protein gene leads to profound suppression of LPS responses ex vivo, whereas in vivo responses remain intact. AB - Gram-negative bacterial lipopolysaccharide (LPS) stimulates phagocytic leukocytes by interacting with the cell surface protein CD14. Cellular responses to LPS are markedly potentiated by the LPS-binding protein (LBP), a lipid-transfer protein that binds LPS aggregates and transfers LPS monomers to CD14. LBP also transfers LPS to lipoproteins, thereby promoting the neutralization of LPS. LBP present in normal plasma has been shown to enhance the LPS responsiveness of cells in vitro. The role of LBP in promoting LPS responsiveness in vivo was tested in LBP deficient mice produced by gene targeting in embryonic stem cells. Whole blood from LBP-deficient animals was 1,000-fold less responsive to LPS as assessed by the release of tumor necrosis factor (TNF)-alpha. Blood from gene-targeted mice was devoid of immunoreactive LBP, essentially incapable of transferring LPS to CD14 in vitro, and failed to support cellular responses to LPS. These activities were restored by the addition of exogenous recombinant murine LBP to the plasma. Despite these striking in vitro findings, no significant differences in TNF-alpha levels were observed in plasma from wild-type and LBP-deficient mice injected with LPS. These data suggest the presence of an LBP-independent mechanism for responding to LPS. These LBP knockout mice may provide a tool for discovering the nature of the presumed second mechanism for transferring LPS to responsive cells. PMID- 9396776 TI - CD4+ T cell help impairs CD8+ T cell deletion induced by cross-presentation of self-antigens and favors autoimmunity. AB - Self-antigens expressed in extrathymic tissues such as the pancreas can be transported to draining lymph nodes and presented in a class I-restricted manner by bone marrow-derived antigen-presenting cells. Such cross-presentation of self antigens leads to CD8+ T cell tolerance induction via deletion. In this report, we investigate the influence of CD4+ T cell help on this process. Small numbers of autoreactive OVA-specific CD8+ T cells were unable to cause diabetes when adoptively transferred into mice expressing ovalbumin in the pancreatic beta cells. Coinjection of OVA-specific CD4+ helper T cells, however, led to diabetes in a large proportion of mice (68%), suggesting that provision of help favored induction of autoimmunity. Analysis of the fate of CD8+ T cells indicated that CD4(+) T cell help impaired their deletion. These data indicate that control of such help is critical for the maintenance of CD8+ T cell tolerance induced by cross-presentation. PMID- 9396777 TI - Resistance to and recovery from lethal influenza virus infection in B lymphocyte deficient mice. AB - In the adaptive immune response to most viruses, both the cellular and humoral arms of the immune system play complementary roles in eliminating virus and virus infected cells and in promoting recovery. To evaluate the relative contribution of CD4+ and CD8+ effector T lymphocytes in virus clearance and recovery, we have examined the host response to lethal type A influenza virus infection in B lymphocyte-deficient mice with a targeted disruption in the immunoglobulin mu heavy chain. Our results indicate that naive B cell-deficient mice have a 50- 100 fold greater susceptibility to lethal type A influenza virus infection than do wild type mice. However, after priming with sublethal doses of influenza, immune B cell-deficient animals show an enhanced resistance to lethal virus infection. This finding indicates that an antibody-independent immune-mediated antiviral mechanism accounts for the increased resistance to lethal virus challenge. To assess the contribution of influenza-specific CD4+ and CD8+ effector T cells in this process, defined clonal populations of influenza-specific CD4+ and CD8+ effector T cells were adoptively transferred into lethally infected B cell deficient mice. Cloned CD8+ effectors efficiently promoted recovery from lethal infection, whereas cloned CD4+ T cells conferred only partial protection. These results suggest that memory T lymphocytes can act independently of a humoral immune response in order to confer resistance to influenza infection in immune individuals. The potential implications of these results for vaccination against human influenza infection are discussed. PMID- 9396778 TI - Itk and Fyn make independent contributions to T cell activation. AB - Itk is a member of the Btk/Tec/Itk family of nonreceptor protein tyrosine kinases (PTKs), and has been implicated in T cell antigen receptor (TCR) signal transduction. Lck and Fyn are the Src-family nonreceptor PTKs that are involved in TCR signaling. To address the question of how these members of different families of PTKs functionally contribute to T cell development and to T cell activation, mice deficient for both Itk and either Lck or Fyn were generated. The Itk/Lck doubly deficient mice exhibited a phenotype similar to that of Lck deficient mice. The phenotype of the Itk/Fyn doubly deficient mice was similar to that of Itk deficient mice. However the Itk/Fyn doubly deficient mice exhibited a more severe defect in TCR-induced proliferation of thymocytes and peripheral T cells than did mice deficient in either kinase alone. These data support the notion that Itk and Fyn both make independent contributions to TCR-induced T cell activation. PMID- 9396779 TI - TRANCE (tumor necrosis factor [TNF]-related activation-induced cytokine), a new TNF family member predominantly expressed in T cells, is a dendritic cell specific survival factor. AB - TRANCE (tumor necrosis factor [TNF]-related activation-induced cytokine) is a new member of the TNF family that is induced upon T cell receptor engagement and activates c-Jun N-terminal kinase (JNK) after interaction with its putative receptor (TRANCE-R). In addition, TRANCE expression is restricted to lymphoid organs and T cells. Here, we show that high levels of TRANCE-R are detected on mature dendritic cells (DCs) but not on freshly isolated B cells, T cells, or macrophages. Signaling by TRANCE-R appears to be dependent on TNF receptor associated factor 2 (TRAF2), since JNK induction is impaired in cells from transgenic mice overexpressing a dominant negative TRAF2 protein. TRANCE inhibits apoptosis of mouse bone marrow-derived DCs and human monocyte-derived DCs in vitro. The resulting increase in DC survival is accompanied by a proportional increase in DC-mediated T cell proliferation in a mixed leukocyte reaction. TRANCE upregulates Bcl-xL expression, suggesting a potential mechanism for enhanced DC survival. TRANCE does not induce the proliferation of or increase the survival of T or B cells. Therefore, TRANCE is a new DC-restricted survival factor that mediates T cell-DC communication and may provide a tool to selectively enhance DC activity. PMID- 9396780 TI - Agents that down-regulate or inhibit protein kinase C circumvent resistance to 1 beta-D-arabinofuranosylcytosine-induced apoptosis in human leukemia cells that overexpress Bcl-2. AB - The effects of the non-tumor-promoting protein kinase C (PKC) activator bryostatin 1 and the PKC inhibitors staurosporine and UCN-01 were examined with respect to modulation of 1-[beta-D-arabinofuranosyl]cytosine (ara-C)-induced apoptosis in human myeloid leukemia cells (HL-60) overexpressing the antiapoptotic protein Bcl-2. HL-60/Bcl-2 cells displayed a 5-fold increase in Bcl 2 protein compared with empty-vector counter-parts (HL-60/pCEP4) but comparable levels of Bax, Mcl-1, and Bcl-xL. After exposure to an equimolar concentration of ara-C (10 microM for 6 hr), HL-60/Bcl-2 cells were significantly less susceptible to apoptosis, DNA fragmentation, and loss of clonogenicity than HL-60/pCEP4 cells. The protective effect of increased Bcl-2 expression was manifested by a failure of ara-C to induce activation/cleavage of the Yama protease (CPP32; caspase-3) and degradation of one of its substrates, poly(ADP-ribose)polymerase to an 85-kDa cleavage product. When HL-60/Bcl-2 cells were preincubated with bryostatin 1 (10 nM; 24 hr) or coincubated with either staurosporine (50 nM; 6 hr) or UCN-01 (300 nM; 6 hr) after a 1-hr preincubation, exposures that exerted minimal effects alone, ara-C-induced apoptosis and DNA fragmentation were restored to levels equivalent to, or greater than, those observed in empty-vector controls. These events were accompanied by restoration of the ability of ara-C to induce CPP32 cleavage and activation, poly(ADP-ribose) polymerase degradation, and inhibition of colony formation. Western analysis of Bcl-2 protein obtained from overexpressing cells treated with bryostatin 1, staurosporine, or UCN-01 revealed the appearance of a slowly migrating species and a general broadening of the protein band, effects that were insensitive to the protein synthesis inhibitor cycloheximide. Alterations in Bcl-2 protein mobility on sodium dodecyl sulfate-polyacrylamide gel electrophoresis were reversed by treatment of lysates with alkaline phosphatase or protein phosphatase 2A; actions of the latter were blocked by the specific phosphatase inhibitor okadaic acid. In vivo labeling studies of Bcl-2 protein demonstrated increased incorporation of [32PO4]orthophosphate in drug-treated cells. Last, phosphorylated Bcl-2 failed to display decreased binding to the proapoptotic protein Bax. Collectively, these findings indicate that bryostatin 1, which down-regulates PKC, and staurosporine and UCN-01, which directly inhibit the enzyme, circumvent resistance of Bcl-2 overexpressing leukemic cells to ara-C-induced apoptosis and activation of the protease cascade. They also raise the possibility that modulation of Bcl-2 phosphorylation status contributes to this effect. PMID- 9396782 TI - Ca2+ feedback on "quantal" Ca2+ release involving ryanodine receptors. AB - The influence of luminal and cytoplasmic Ca2+ on the ability of ryanodine sensitive stores to undergo multiple partial ("quantal") releases has been assessed. Increased luminal Ca2+ levels do indeed modulate sarcoplasmic reticulum Ca2+ release by lowering the threshold agonist concentration required to elicit release, but the decrease in luminal Ca2+ that accompanies a partial release is not sufficient by itself to terminate release. Similarly, an increase in cytoplasmic Ca2+ lowers the threshold agonist concentration required to elicit release; thus, the bulk cytoplasmic Ca2+ levels attained during a release would only stimulate further release, not terminate it before it reached completion. Very high cytoplasmic Ca2+ levels (1-3 mM) also triggered release but were unable to terminate release before reaching completion. Thus, even the high local cytoplasmic Ca2+ concentration that might accompany release would also not terminate release. It is concluded that Ca2+ feedback can modulate release through ryanodine receptors but that it does not account for the properties of quantal release. The low affinity inhibitor tetracaine induces a decrease in the extent of release that cannot be explained solely by heterogeneous caffeine sensitivity of the stores. The results are interpreted in terms of a scheme that includes (i) heterogeneous sensitivity of stores, conferred in part by differences in luminal Ca2+ content and (ii) adaptive behavior on the part of individual ryanodine receptors. PMID- 9396781 TI - Distinct sites for inverse modulation of N-methyl-D-aspartate receptors by sulfated steroids. AB - Steroid sulfation occurs in nervous tissue and endogenous sulfated steroids can act as positive or negative modulators of N-methyl-D-aspartate (NMDA) receptor function. In the current study, structure-activity relationships for sulfated steroids were examined in voltage-clamped chick spinal cord and rat hippocampal neurons in culture and in Xenopus laevis oocytes expressing NR1(100) and NR2A subunits. The ability of pregnenolone sulfate (a positive modulator) and epipregnanolone sulfate (a negative modulator) to compete with each another, as well as with other known classes of NMDA receptor modulators, was examined. The results show that steroid positive and negative modulators act at specific, extracellularly directed sites that are distinct from one another and from the spermine, redox, glycine, Mg2+, MK-801, and arachidonic acid sites. Sulfated steroids are effective as modulators of ongoing glutamate-mediated synaptic transmission, which is consistent with their possible role as endogenous neuromodulators in the CNS. PMID- 9396783 TI - KvLQT1 potassium channel but not IsK is the molecular target for trans-6-cyano-4 (N-ethylsulfonyl-N-methylamino)-3-hydroxy-2,2-dimethyl- chromane. AB - Mutations in the KvLQT1 gene are the cause for the long QT syndrome [Circulation 94:1996-2012 (1996)]. Coexpression of KvLQT1 in association with the channel regulator protein IsK produces a K+ current with characteristics reminiscent of the slow component of the delayed rectifier in cardiac myocytes. We explored the pharmacological properties of trans-6-cyano-4-(N-ethylsulfonyl-N-methylamino)-3 hydroxy-2,2-dime thyl- chromane (293B), a chromanol compound, on the K+ current produced by direct intranuclear injection of KvLQT1 and IsK cDNA plasmids in COS 7 cells. Injected cells were recorded by means of the whole-cell and cell attached patch-clamp configurations under chloride-free conditions. Cells injected with KvLQT1 cDNA alone exhibited a fast-activating outward K+ current, whereas cells coinjected with KvLQT1 plus IsK cDNAs exhibited a time-dependent outward current with slower activation kinetics. The chromanol 293B blocked the K+ current related to KvLQT1 expression in both the absence or presence of IsK. The IC50 value for 293B to block KvLQT1-related current was not significantly modified by the presence of IsK (9.9 microM in the absence of IsK versus 9.8 microM in its presence). The block produced by 293B was strongly voltage dependent inasmuch as it was close to 0 at -80 mV and occurred during a depolarizing voltage step. The time constants for the drug to block the current were in the same order of magnitude as activation kinetics of the current. Kinetics for drug unblock at the holding potential were much faster, in the order of a few tenths of a msec. KvLQT1 currents recorded in the cell-attached configuration were also blocked by externally applied 293B, suggesting that the compound penetrated the cell to block the channel. Cromakalim, another chromanol compound, also blocked KvLQT1 currents. Our results show that the chromanol compound 293B is targeted to KvLQT1 channels but not to the IsK regulator. PMID- 9396784 TI - Constitutive activity of a chimeric D2/D1 dopamine receptor. AB - Chimeric D1/D2 receptors were constructed to identify structural determinants of drug affinity and efficacy. We previously reported that chimeras that had D1 receptor transmembrane domain VII together with amino-terminal sequence from the D2 receptor were nonfunctional. D2/D1 chimeras were constructed that contained D2 receptor sequence at the amino- and carboxyl-terminal ends and D1 receptor sequence in the intervening region. Chimeric receptors with D2 sequence from transmembrane domain 7 to the carboxyl terminus together with D2 receptor sequence from the amino terminus through transmembrane helix 4 (D2[1-4,7]) and 5 (D2[1-5,7]) bound [3H]spiperone with high affinity, consistent with the hypothesis that D2 receptor transmembrane domain I or II is incompatible with D1 receptor transmembrane domain VII. D2[1-4,7] and D2[1-5,7] had affinities similar to D1 and D2 receptors for most nonselective dopamine antagonists and had affinities for most of the selective antagonists that were intermediate between those of the parent receptors. D2[1-4,7] and D2[1-5,7] mediated dopamine receptor agonist-induced stimulation and inhibition, respectively, of cAMP accumulation. The more efficient coupling of D2[1-5,7] to inhibition of cAMP accumulation, compared with the coupling of D2[5-7] and D2[3-7], supports the view that multiple D2 receptor cytoplasmic domains acting in concert are necessary for receptor activation of Gi. In contrast, D2[1-4,7], which contains only one cytoplasmic loop (the third) from the D1 receptor, is capable of activating Gs. D2[1-4,7] exhibited several characteristics of a constitutively active receptor, including enhanced basal (unliganded) stimulation of cAMP accumulation, high affinity for agonists even in the presence of GTP, and blunted agonist-stimulated cAMP accumulation. A number of dopamine receptor antagonists were inverse agonists at D2[1-4,7], inhibiting basal cAMP accumulation. Some of these drugs were also inverse agonists at the D1 receptor. Interestingly, several antagonists also potentiated forskolin-stimulated cAMP accumulation via D2[1-5,7] and via the D2 receptor, which could reflect inverse agonist inhibition of native constitutive activity of this receptor. PMID- 9396785 TI - Differences in agonist/antagonist binding affinity and receptor transduction using recombinant human gamma-aminobutyric acid type A receptors. AB - Using human gamma-aminobutyric acid type A (GABAA) receptor subunit combinations, expressed in cell lines and Xenopus laevis oocytes, the pharmacology of a number of ligands interacting directly with the GABA recognition site has been studied in [3H]muscimol binding and electrophysiologically. The binding affinity of GABAA agonist and antagonist ligands showed small but statistically significant dependence on the subunit composition of receptors that include gamma 2 and different alpha and beta subunits. The potency of antagonist ligands was largely independent of receptor subunit composition, whereas the composition of receptors expressed in oocytes strongly influenced the EC50 value of agonists. An apparent reciprocal correlation between subunits favoring agonist binding and antagonist binding, respectively, was observed. Whereas antagonists showed comparable potencies in binding and functional studies, the potency of agonists in binding studies was generally two to three orders of magnitude higher than the agonist potencies measured electrophysiologically. 5-(4-Piperidyl)isothiazol-3-ol, which behaves as a low efficacy partial agonist at GABAA receptors in cultured cortical neurons, showed no efficacy in oocytes, but produced pure antagonist effects with a binding/functional affinity ratio between those observed for the agonists and antagonists. It is concluded that the GABAA receptor mechanisms transducing binding into physiological response, but not the binding per se, is dependent on the receptor subunit composition. PMID- 9396786 TI - S-adenosylhomocysteine hydrolase inhibitors interfere with the replication of human immunodeficiency virus type 1 through inhibition of the LTR transactivation. AB - Various analogues of adenosine have been described as inhibitors of S adenosylhomocysteine (AdoHcy) hydrolase, and some of these AdoHcy hydrolase inhibitors (e.g., 3-deazaadenosine, 3-deazaaristeromycin, and 3-deazaneplanocin A) have also been reported to inhibit the replication of human immunodeficiency virus type 1 (HIV-1). When evaluated against HIV-1 replication in MT-4 cells, macrophages, or phytohemagglutinin-stimulated peripheral blood lymphocytes infected acutely or chronically with HIV-1IIIB or HIVBaL strains, a wide range of adenosine analogues did not inhibit HIV-1IIIB replication for 50% at subtoxic concentrations. However, they inhibited HIV-1 replication in HeLa CD4+ LTR-LacZ cells at concentrations well below cytotoxicity threshold. A close correlation was found among the inhibitory effect of the compounds on AdoHcy hydrolase activity, their inhibition of HIV-1 replication in Hela CD4+ LTR-LacZ cells, and their inhibition of the HIV-1 Tat-dependent and -independent transactivation of the long terminal repeat, whereas no inhibitory effect was seen on HIV-1 reverse transcription or a Tat-independent cytomegalovirus promoter. Our results suggest that AdoHcy hydrolase and the associated S-adenosylmethionine-dependent methylation mechanism play a role in the process of long terminal repeat transactivation and, hence, HIV replication. PMID- 9396787 TI - Expression cloning and receptor pharmacology of human calcitonin receptors from MCF-7 cells and their relationship to amylin receptors. AB - Human breast cell carcinoma MCF-7 cells were found to bind 125I-labeled rat amylin (rAmylin) and the peptide amylin antagonist radioligand 125I-AC512 with high affinity. This high affinity binding possessed characteristics unique to the already defined high affinity binding site for amylin in the rat nucleus accumbens [Mol. Pharmacol. 44:493-497 (1993); J. Pharmacol. Exp. Ther. 270:779 787 (1994); Eur. J. Pharmacol. 262:133-141 (1994)]. To further define this receptor, we report results of expression cloning studies from an MCF-7 cell library. We isolated two variants of a seven-transmembrane receptor that were identical to two previously described human calcitonin receptors (hCTR1 and hCTR2). These receptors were characterized by expression in different surrogate host cell systems. Transient expression of hCTR1 in COS cells yielded membranes that bound 125I-AC512 and 125I-salmon calcitonin with high affinity, but no high affinity binding was observed with 125I-human calcitonin (hCAL) or 125I-rAmylin. Stable expression of hCTR1 in HEK 293 cells produced similar data. In contrast, expression of hCTR2 in COS cells yielded membranes that bound 125I-AC512, 125I hCAL, and 125I-rAmylin with high affinity. The agonists 125I-hCAL and 125I rAmylin bound 65% and 1.5%, respectively, of the sites bound by the antagonist radioligand 125I-AC512 in this expression system. This pattern of binding was repeated in HEK 293 cells stably transfected with hCTR2 (125I-hCAL = 24.8% Bmax, 125I-rAmylin = 8% Bmax). In both expression systems, the agonists hCAL and rAmylin were much more potent in displacing their radioligand counterparts than was the antagonist radioligand 125I-AC512. For example, the pKi value for displacement of 125I-AC512 by rAmylin was 7.2 in HEK 293 cells but rose to 9.1 when displacing 125I-rAmylin. Finally, hCTR2 was expressed in baculovirus infected Ti ni cells. In this system, only specific binding to the antagonist 125I-AC512 and agonist 125I-hCAL was observed; no binding to 125I-rAmylin could be detected. These data are discussed in terms of two working hypotheses. The first is that amylin is a weak agonist for hCTR2 and that this receptor is unrelated to the amylin receptor found in this cell line. The second is that hCTR2 couples to different G proteins for calcitonin and amylin function in different cells. At present, these data cannot be used to disprove conclusively either hypothesis. PMID- 9396788 TI - A multiplicity of mediators: alternative forms of transcription complexes communicate with transcriptional regulators. AB - The already complex process of transcription by RNA polymerase II has become even more complicated in the last few years with the identification of auxiliary factors in addition to the essential general initiation factors. In many cases these factors, which have been termed mediators or co-activators, are only required for activated or repressed transcription. In some cases the effects are specific for certain activators and repressors. Recently some of these auxiliary factors have been found in large complexes with either TBP, as TBP-associated factors (TAFs) in the general factor TFIID, or with pol II and a subset of the general factors, referred to as the 'holoenzyme'. Although the exact composition of these huge assemblies is still a matter of some debate, it is becoming clear that the complexes themselves come in more than one form. In particular, at least four forms of TFIID have been described, including one that contains a tissue specific TAF and another with a cell type-specific form of TBP. In addition, in yeast there are at least two forms of the 'holoenzyme' distinguished by their mediator composition and by the spectrum of transcripts whose expression they affect. Genetic and biochemical analyses have begun to identify the interactions between the components of these complexes and the ever increasing family of DNA binding regulatory factors. These studies are complicated by the fact that individual regulatory factors often appear to have redundant interactions with multiple mediators. The existence of these different forms of transcription complexes defines a new target for regulation of subsets of eukaryotic genes. PMID- 9396789 TI - Template directed incorporation of nucleotide mixtures using azole-nucleobase analogs. AB - DNA that encodes elements for degenerate replication events by use of artificial nucleobases offers a versatile approach to manipulating sequences for applications in biotechnology. We have designed a family of artificial nucleobases that are capable of assuming multiple hydrogen bonding orientations through internal bond rotations to provide a means for degenerate molecular recognition. Incorporation of these analogs into a single position of a PCR primer allowed for analysis of their template effects on DNA amplification catalyzed by Thermus aquaticus (Taq) DNA polymerase. All of the nucleobase surrogates have similar shapes but differ by structural alterations that influence their electronic character. These subtle distinctions were able to influence the Taq DNA polymerase dependent incorporation of the four natural deoxyribonucleotides and thus, significantly expand the molecular design possibilities for biochemically functional nucleic acid analogs. PMID- 9396790 TI - A new nomenclature for the cytoplasmic ribosomal proteins of Saccharomyces cerevisiae. AB - The availability of the complete sequence of the Saccharomyces cerevisiae genome has allowed a comprehensive analysis of the genes encoding cytoplasmic ribosomal proteins in this organism. On the basis of this complete inventory a new nomenclature for the yeast ribosomal proteins is presented. PMID- 9396791 TI - The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. AB - CLUSTAL X is a new windows interface for the widely-used progressive multiple sequence alignment program CLUSTAL W. The new system is easy to use, providing an integrated system for performing multiple sequence and profile alignments and analysing the results. CLUSTAL X displays the sequence alignment in a window on the screen. A versatile sequence colouring scheme allows the user to highlight conserved features in the alignment. Pull-down menus provide all the options required for traditional multiple sequence and profile alignment. New features include: the ability to cut-and-paste sequences to change the order of the alignment, selection of a subset of the sequences to be realigned, and selection of a sub-range of the alignment to be realigned and inserted back into the original alignment. Alignment quality analysis can be performed and low-scoring segments or exceptional residues can be highlighted. Quality analysis and realignment of selected residue ranges provide the user with a powerful tool to improve and refine difficult alignments and to trap errors in input sequences. CLUSTAL X has been compiled on SUN Solaris, IRIX5.3 on Silicon Graphics, Digital UNIX on DECstations, Microsoft Windows (32 bit) for PCs, Linux ELF for x86 PCs, and Macintosh PowerMac. PMID- 9396792 TI - Role of acceptor stem conformation in tRNAVal recognition by its cognate synthetase. AB - Although the anticodon is the primary element in Escherichia coli tRNAValfor recognition by valyl-tRNA synthetase (ValRS), nucleotides in the acceptor stem and other parts of the tRNA modulate recognition. Study of the steady state aminoacylation kinetics of acceptor stem mutants of E.coli tRNAValdemonstrates that replacing any base pair in the acceptor helix with another Watson-Crick base pair has little effect on aminoacylation efficiency. The absence of essential recognition nucleotides in the acceptor helix was confirmed by converting E.coli tRNAAlaand yeast tRNAPhe, whose acceptor stem sequences differ significantly from that of tRNAVal, to efficient valine acceptors. This transformation requires, in addition to a valine anticodon, replacement of the G:U base pair in the acceptor stem of these tRNAs. Mutational analysis of tRNAValverifies that G:U base pairs in the acceptor helix act as negative determinants of synthetase recognition. Insertion of G:U in place of the conserved U4:A69 in tRNAValreduces the efficiency of aminoacylation, due largely to an increase in K m. A smaller but significant decline in aminoacylation efficiency occurs when G:U is located at position 3:70; lesser effects are observed for G:U at other positions in the acceptor helix. The negative effects of G:U base pairs are strongly correlated with changes in helix structure in the vicinity of position 4:69 as monitored by19F NMR spectroscopy of 5-fluorouracil-substituted tRNAVal. This suggests that maintaining regular A-type RNA helix geometry in the acceptor stem is important for proper recognition of tRNAValby valyl-tRNA synthetase.19F NMR also shows that formation of the tRNAVal-valyl-tRNA synthetase complex does not disrupt the first base pair in the acceptor stem, a result different from that reported for the tRNAGln-glutaminyl-tRNA synthetase complex. PMID- 9396793 TI - Recognition of GC base pairs by triplex forming oligonucleotides containing nucleosides derived from 2-aminopyridine. AB - We have attempted to alleviate the pH dependency of triplex recognition of guanine by using intermolecular triplexes containing 2-amino-5-(2-deoxy-d ribofuranosyl)pyridine (AP) as an analogue of 2'-deoxycytidine (dC). We find that for the beta-anomer of AP, the complex between (AP)6T6and the target site G6A6*T6C6is stable, generating a clear DNase I footprint at oligonucleotide concentrations as low as 0.25 microM at pH 5.0, in contrast to 50 microM C6T6which has no effect on the cleavage pattern. This complex is still stable at pH 6.5 producing a footprint with 1 microM oligonucleotide. Oligonucleotides containing the alpha-anomer of AP are much less effective than the beta-anomer, though in some instances they are more stable than the unmodified oligonucleotides. The results of molecular dynamics studies on a range of AP containing triplexes has rationalized the observed stability behaviour in terms of hydrogen-bonding behaviour. PMID- 9396794 TI - Mirror image alternative interaction patterns of the same tRNA with either class I arginyl-tRNA synthetase or class II aspartyl-tRNA synthetase. AB - Gene cloning, overproduction and an efficient purification protocol of yeast arginyl-tRNA synthetase (ArgRS) as well as the interaction patterns of this protein with cognate tRNAArgand non-cognate tRNAAspare described. This work was motivated by the fact that the in vitro transcript of tRNAAspis of dual aminoacylation specificity and is not only aspartylated but also efficiently arginylated. The crystal structure of the complex between class II aspartyl-tRNA synthetase (AspRS) and tRNAAsp, as well as early biochemical data, have shown that tRNAAspis recognized by its variable region side. Here we show by footprinting with enzymatic and chemical probes that transcribed tRNAAspis contacted by class I ArgRS along the opposite D arm side, as is homologous tRNAArg, but with idiosyncratic interaction patterns. Besides protection, footprints also show enhanced accessibility of the tRNAs to the structural probes, indicative of conformational changes in the complexed tRNAs. These different patterns are interpreted in relation to the alternative arginine identity sets found in the anticodon loops of tRNAArgand tRNAAsp. The mirror image alternative interaction patterns of unmodified tRNAAspwith either class I ArgRS or class II AspRS, accounting for the dual identity of this tRNA, are discussed in relation to the class defining features of the synthetases. This study indicates that complex formation between unmodified tRNAAspand either ArgRS and AspRS is solely governed by the proteins. PMID- 9396795 TI - Electric field directed nucleic acid hybridization on microchips. AB - Selection and adjustment of proper physical parameters enables rapid DNA transport, site selective concentration, and accelerated hybridization reactions to be carried out on active microelectronic arrays. These physical parameters include DC current, voltage, solution conductivity and buffer species. Generally, at any given current and voltage level, the transport or mobility of DNA is inversely proportional to electrolyte or buffer conductivity. However, only a subset of buffer species produce both rapid transport, site specific concentration and accelerated hybridization. These buffers include zwitterionic and low conductivity species such as: d- and l-histidine; 1- and 3 methylhistidines; carnosine; imidazole; pyridine; and collidine. In contrast, buffers such as glycine, beta-alanine and gamma-amino-butyric acid (GABA) produce rapid transport and site selective concentration but do not facilitate hybridization. Our results suggest that the ability of these buffers (histidine, etc.) to facilitate hybridization appears linked to their ability to provide electric field concentration of DNA; to buffer acidic conditions present at the anode; and in this process acquire a net positive charge which then shields or diminishes repulsion between the DNA strands, thus promoting hybridization. PMID- 9396796 TI - CDF-1, a novel E2F-unrelated factor, interacts with cell cycle-regulated repressor elements in multiple promoters. AB - The cdc25C , cdc2 and cyclin A promoters are controlled by transcriptional repression through two contiguous protein binding sites, termed the CDE and CHR. In the present study we have identified a factor, CDF-1, which interacts with the cdc25C CDE-CHR module. CDF-1 binds to the CDE in the major groove and to the CHR in the minor grove in a cooperative fashion in vitro , in a manner similar to that seen by genomic footprinting. In agreement with in vivo binding data and its putative function as a periodic repressor, DNA binding by CDF-1 in nuclear extracts is down-regulated during cell cycle progression. CDF-1 also binds avidly to the CDE-CHR modules of the cdc2 and cyclin A promoters, but not to the E2F site in the B- myb promoter. Conversely, E2F complexes do not recognize the cdc25C CDE-CHR and CDF-1 is immunologically unrelated to all known E2F and DP family members. This indicates that E2F- and CDF-mediated repression is controlled by different factors acting at different stages during the cell cycle. While E2F-mediated repression seems to be associated with genes that are up regulated early (around mid G1), such as B- myb , CDE-CHR-controlled genes, such as cdc25C , cdc2 and cyclin A , become derepressed later. Finally, the fractionation of native nuclear extracts on glycerol gradients leads to separation of CDF-1 from both E2F complexes and pocket proteins of the pRb family. This emphasizes the conclusion that CDF-1 is not an E2F family member and points to profound differences in the cell cycle regulation of CDF-1 and E2F. PMID- 9396797 TI - The differential binding of E2F and CDF repressor complexes contributes to the timing of cell cycle-regulated transcription. AB - B- myb and cdc25C exemplify different groups of genes whose transcription is consecutively up-regulated during the cell cycle. Both promoters are controlled by transcriptional repression via modules consisting of an E2F binding site (E2FBS) or the related CDE plus a contiguous CHR co-repressor element. We now show that the B- myb repressor module, which is derepressed early (mid G1), is preferentially recognized by E2F-DP complexes and that a mutation selectively abolishing E2F binding impairs regulation. In contrast, the cdc25C repressor module, which is derepressed late (S/G2), interacts selectively with CDE-CHR binding factor-1 (CDF-1). E2F binding, but not CDF-1 binding, requires specific nucleotides flanking the E2FBS/CDE core, while CDF-1 binding, but not E2F binding, depends on specific nucleotides in the CHR. Swapping these nucleotides between the two promoters profoundly changes protein binding patterns and alters expression kinetics. Thus predominant CDF-1 binding leads to derepression in late S, predominant E2F binding results in up-regulation in late G1, while promoters binding both E2F and CDF-1 with high efficiency show intermediate kinetics. Our results support a model where the differential binding of E2F and CDF-1 repressor complexes contributes to the timing of promoter activity during the cell cycle. PMID- 9396798 TI - CDF-1-mediated repression of cell cycle genes targets a specific subset of transactivators. AB - The cdc25C , cyclin A and cdc2 genes are regulated during the cell cycle through two contiguous repressor binding sites, the CDE and CHR, located in the region of transcription initiation and interacting with a factor termed CDF-1. The target of this repression seems to be transcriptional activation of these promoters by transcription factors bound upstream. The majority of these factors falls into the class of glutamine-rich activators, suggesting that CDF-1-mediated repression might be activation domain specific. In the present study we have used chimeric promoter constructs to demonstrate that the cdc25C UAS, but not the core promoter, is crucial for repression. In addition, we show that only specific transcription factors and activation domains are responsive to CDE-CHR-mediated cell cycle regulation. These observations clearly indicate that CDF-1 interferes with activation of transcription by a specific subset of transactivators. The repressible activation domains belong to the same class of glutamine-rich activators, pointing to specific interactions of CDF-1 with components of the transcription machinery. In agreement with this conclusion we find that a simple inversion of the CDE-CHR module completely abrogates cell cycle-regulated repression. PMID- 9396799 TI - Characterization of the TATA-less core promoter of the cell cycle-regulated cdc25C gene. AB - The TATA- and Inr-less promoter of the human cdc25C gene is regulated during the cell cycle through binding of a repressor to two contiguous promoter-proximal elements, the CDE and CHR. In this study we have characterized in detail the region of the cdc25C promoter immediately downstream of these elements. Several lines of evidence suggest that this region of approximately 60 bp acts as the core promoter. This sequence: (i) harbors most of the transcription initiation sites; (ii) possesses basal promoter activity in vivo ; (iii) shows no stable protein binding in vivo as indicated by genomic dimethyl sulfate and phenanthroline copper footprinting; (iv) contains single-stranded regions in vivo as shown by potassium permanganate footprinting; (v) is hypersensitive to DNase I cleavage in permeabilized cells. Mutational analysis of the core promoter revealed the presence of three sites which play a role in transcription. Two of these sites were found to represent low affinity binding sites for transcription factors of the Sp1 family. Mutation of these sites led to decreased levels of transcription, while their alteration to canonical Sp1 sites impaired cell cycle regulation. Thus the transient interaction of Sp1 with the core promoter appears to be necessary for maximal transcription without perturbing cell cycle regulation. PMID- 9396800 TI - NUCPLOT: a program to generate schematic diagrams of protein-nucleic acid interactions. AB - Proteins that bind to DNA are found in all areas of genetic activity within the cell. To help understand how these proteins perform their various functions, it is useful to analyse which residues are involved in binding to the DNA and how they interact with the bases and sugar-phosphate backbone of nucleic acids. Here we describe a program called NUCPLOT which can automatically identify these interactions from the 3D atomic coordinates of the complex from a PDB file and generate a plot that shows all the interactions in a schematic manner. The program produces a PostScript output file representing hydrogen, van der Waals and covalent bonds between the protein and the DNA. The resulting diagram is both clear and simple and allows immediate identification of important interactions within the structure. It also facilitates comparison of binding found in different structures. NUCPLOT is a completely automatic program, which can be used for any protein-DNA complex and will also work for certain protein-RNA structures. PMID- 9396802 TI - Inhibitory properties of double-helix-forming circular oligonucleotides. AB - Several circular oligonucleotides were synthesized and characterized by electrospray ionization mass spectrometry. Experiments on termination of primer extension catalysed by DNA polymerases, Klenow fragment and Tth have demonstrated that a double helix forming circular 2'-deoxyribooligomer containing a 25mer sequence complementary to the target single-stranded DNA along with a 34mer random mismatching stretch appears to be a potent inhibitor of replication in vitro. Studies on inhibition of luciferase gene expression in a cell-free transcription-translation system have shown that a duplex forming circular 2' deoxyribooligonucleotide containing a 25mer sequence complementary to the target mRNA and a 14mer random mismatching stretch can serve as an effective antisense compound as a standard linear complementary oligomer. Features of double helix forming circular oligonucleotides composed of 2'-deoxyribonucleosides seem to be useful for the design of new antigene and antisense agents. PMID- 9396801 TI - A novel nucleic acid-binding protein that interacts with human rad51 recombinase. AB - Using the yeast two-hybrid system, we isolated a cDNA encoding a novel human protein, named Pir51, that strongly interacts with human Rad51 recombinase. Analysis in vitro confirmed the interaction between Rad51 and Pir51. Pir51 mRNA is expressed in a number of human organs, most notably in testis, thymus, colon and small intestine. The Pir51 gene locus was mapped to chromosome 12p13.1-13. 2 by fluorescence in situ hybridization. The Pir51 protein was expressed in Escherichia coli and purified to near homogeneity. Biochemical analysis shows that the Pir51 protein binds both single- and double-stranded DNA, and is capable of aggregating DNA. The protein also binds RNA. The Pir51 protein may represent a new member of the multiprotein complexes postulated to carry out homologous recombination and DNA repair in mammalian cells. PMID- 9396803 TI - Interaction of minor groove binding ligands with long AT tracts. AB - We have used quantitative DNase I footprinting to examine the ability of distamycin and Hoechst 33258 to discriminate between different arrangements of AT residues, using synthetic DNA fragments containing multiple blocks of (A/T)6or (A/T)10in identical sequence environments. Previous studies have shown that these ligands bind less well to (A/T)4sites containing TpA steps. We find that in (A/T)6tracts distamycin shows little discrimination between the various sites, binding approximately 2-fold stronger to TAATTA than (TA)3, T3A3and GAATTC. In contrast, Hoechst 33258 binds approximately 20-fold more tightly to GAATTC and TAATTA than T3A3and (TA)3. Hydroxyl radical footprinting reveals that both ligands bind in similar locations at the centre of each AT tract. At (A/T)10sites distamycin binds with similar affinity to T5A5, (TA)5and AATT, though bands in the centre of (TA)5are protected at approximately 50-fold lower concentration than those towards the edges. Hoechst 33258 shows a similar pattern of preference, with strong binding to AATT, T5A5and the centre of (TA)5. Hydroxyl radical footprinting reveals that at low concentrations both ligands bind at the centre of (TA)5and A5T5, while at higher concentrations ligand molecules bind to each end of the (A/T)10tracts. At T5A5two ligand molecules bind at either end of the site, even at the lowest ligand concentration, consistent with the suggestion that these compounds avoid the TpA step. Similar DNase I footprinting experiments with a DNA fragment containing T n (n = 3-6) tracts reveals that both ligands bind in the order T3< T4 << T5 = T6. PMID- 9396804 TI - A structure-specific endonuclease from cauliflower (Brassica oleracea var. botrytis) inflorescence. AB - A protein with structure-specific endonuclease activity has been purified to near homogeneity from cauliflower ( Brassica oleracea var. botrytis) inflorescence through five successive column chromatographies. The protein is a single polypeptide with a molecular mass of 40 kDa. Using three different branched DNA structures (flap, pseudo-Y and stem-loop) we found that the enzyme, a cauliflower structure-specific endonuclease, cleaved the single-stranded tail in the 5'-flap and 5'-pseudo-Y structures, whereas it could not incise the 3'-flap and 3'-pseudo Y structures. The incision points occur around the single strand-duplex junction in these DNA substrates and the enzyme leaves 5'-PO4 and 3'-OH termini on DNA. The protein also endonucleolytically cleaves on the 3'-side of the single stranded region at the junction of unpaired and duplex DNA in the stem-loop structure. The structure-specific endonuclease activity is stimulated by Mg2+ and by Mn2+, but not by Ca2+. Like mammalian FEN-1, the protein has weak 5'-->3' double-stranded DNA-specific exonuclease activity. These results indicate that the cauliflower protein is a plant structure-specific endonuclease like mammalian FEN-1 or may be the plant alternative. PMID- 9396805 TI - The trypanosomatid Leptomonas collosoma 7SL RNA gene. Analysis of elements controlling its expression. AB - We have previously reported the co-purification of a tRNA-like molecule with the Trypanosoma brucei SRP complex [Beja et al . (1993) Mol. Biochem. Parasitol . 57, 223-230]. To examine whether the trypanosome SRP has a unique composition compared with that of other eukaryotes, we analyzed the 7SL RNA and the SRP complex of the monogenetic trypanosomatid Leptomonas collosoma. The 7SL RNA from L. collosoma was cloned, and its gene was sequenced. In contrast to T. brucei , two 7SL RNA transcripts were detected in L.collosoma that originate from a single copy gene. Using stable cell lines expressing tagged 7SL RNA, we demonstrate that the tRNAArggene located 98 bp upstream to the 7SL RNA serves as part of the 7SL RNA extragenic promoter. The steady-state level of 7SL RNA was found to be tightly regulated, since the presence of the gene on the multi-copy plasmid repressed the synthesis of the chromosomal gene. Cell lines carrying truncated 7SL RNA genes were established and their expression indicated that domain I is essential for expressing the 7SL RNA. No constructs carrying portions of the 7SL RNA were expressed, except for a construct which lacked 23 nt from the 3'end of the RNA. This suggests that 90% of the 7SL RNA molecule is important for its maintenance as a stable small RNA. We propose that the repression phenomenon may originate from a regulatory mechanism that coordinates the level of the 7SL RNA by its binding proteins. PMID- 9396806 TI - The effect of structure in a long target RNA on ribozyme cleavage efficiency. AB - Inhibition of gene expression by catalytic RNA (ribozymes) requires that ribozymes efficiently cleave specific sites within large target RNAs. However, the cleavage of long target RNAs by ribozymes is much less efficient than cleavage of short oligonucleotide substrates because of higher order structure in the long target RNA. To further study the effects of long target RNA structure on ribozyme cleavage efficiency, we determined the accessibility of seven hammerhead ribozyme cleavage sites in a target RNA that contained human immunodeficiency virus type 1 (HIV-1) vif - vpr . The base pairing-availability of individual nucleotides at each cleavage site was then assessed by chemical modification mapping. The ability of hammerhead ribozymes to cleave the long target RNA was most strongly correlated with the availability of nucleotides near the cleavage site for base pairing with the ribozyme. Moreover, the accessibility of the seven hammerhead ribozyme cleavage sites in the long target RNA varied by up to 400 fold but was directly determined by the availability of cleavage sites for base pairing with the ribozyme. It is therefore unlikely that steric interference affected hammerhead ribozyme cleavage. Chemical modification mapping of cleavage site structure may therefore provide a means to identify efficient hammerhead ribozyme cleavage sites in long target RNAs. PMID- 9396808 TI - Differential effect of H1 variant overproduction on gene expression is due to differences in the central globular domain. AB - The in vivo overproduction of two mouse histone H1 variants in homologous mouse fibroblasts has opposite effects on gene expression. Overproduction of H1(0) results in repression of transcript levels of all polymerase II genes tested. In contrast, overproduction of H1c results in elevated levels of transcripts. We created a series of chimeric H1 genes in which the regions encoding the three structural domains common to this family of these proteins were systematically switched. Overexpression of these genes in vivo resulted in the accumulation of large amounts of the chimeric H1 in chromatin. Analysis of the effects of overproduction of these proteins revealed that the differential effect of H1 variant overproduction on gene expression is due to differences in the central globular domain. PMID- 9396807 TI - Information analysis of Fis binding sites. AB - Originally discovered in the bacteriophage Mu DNA inversion system gin, Fis (Factor for Inversion Stimulation) regulates many genetic systems. To determine the base frequency conservation required for Fis to locate its binding sites, we collected a set of 60 experimentally defined wild-type Fis DNA binding sequences. The sequence logo for Fis binding sites showed the significance and likely kinds of base contacts, and these are consistent with available experimental data. Scanning with an information theory based weight matrix within fis, nrd, tgt/sec and gin revealed Fis sites not previously identified, but for which there are published footprinting and biochemical data. DNA mobility shift experiments showed that a site predicted to be 11 bases from the proximal Salmonella typhimurium hin site and a site predicted to be 7 bases from the proximal P1 cin site are bound by Fis in vitro. Two predicted sites separated by 11 bp found within the nrd promoter region, and one in the tgt/sec promoter, were also confirmed by gel shift analysis. A sequence in aldB previously reported to be a Fis site, for which information theory predicts no site, did not shift. These results demonstrate that information analysis is useful for predicting Fis DNA binding. PMID- 9396809 TI - A rapid in vitro method for obtaining RNA accessibility patterns for complementary DNA probes: correlation with an intracellular pattern and known RNA structures. AB - A technique is described to identify the rare sequences within an RNA molecule that are available for efficient interaction with complementary DNA probes: the target RNA is digested by RNase H in the presence of a random pool of complementary DNA fragments generated from the same DNA preparation that was used for target RNA synthesis. The DNA region was amplified by PCR, partially digested with DNase and denatured prior to RNA binding. In the presence of single-stranded DNA fragments the RNA was digested with RNase H such that, on average, each molecule was cut once. Cleavage sites were detected by gel electrophoresis either directly with end-labeled RNA or by primer extension. The pattern of accessible sites on c- raf mRNA was determined and compared with the known profile of activity of oligonucleotides found in cells, showing the merit of the method for predicting oligonucleotides which are efficient for in vivo antisense targeting. New susceptible sites in the 3'-untranslated region of c- raf mRNA were identified. Also, four RNAs were probed to ascertain to what extent structure predicts accessibility: the P4-P6 domain of the Tetrahymena group I intron, yeast tRNAAsp, Escherichia coli tmRNA and a part of rat 18S rRNA. PMID- 9396810 TI - Effects of heterologous downstream sequences on the activity of the HIV-1 promoter and its response to Tat. AB - In HIV-1 infection, Tat acts at least in part to control transcriptional elongation by overcoming premature transcriptional termination. In some other genes this process is governed by DNA elements called attenuators in concert with cellular transcription factors. To understand the action of Tat more fully and explore its role as an anti-attenuator, we examined the ability of several natural and synthetic attenuation sequences to modulate transcription initiated at the HIV LTR. Fragments containing these signals were inserted downstream of the TAR element in an HIV-CAT chimera and their effects on transcription were assessed both in vitro and in vivo. Runoff transcription assays in HeLa cell extracts demonstrated that the attenuators give rise to premature termination of transcripts initiated from the heterologous HIV-LTR promoter in vitro. When transiently expressed following transfection into Cos cells, however, premature transcript termination at the attenuation site was not observed. Nevertheless, many of the inserted sequences exerted marked effects on CAT gene expression and on transactivation by Tat at both the RNA and protein levels. The nature and magnitude of the effects depended upon the identity of the attenuator and its orientation but only one of 16 sequences tested met the criteria for a Tat suppressible attenuator in vivo. One other sequence, in contrast, severely reduced Tat-activated transcription without inhibiting basal transcription These results indicate that sequences downstream of the HIV LTR can influence its function as a promoter and its response to Tat transactivation, but lend little support to their role as attenuators in vivo. PMID- 9396811 TI - HUB1, a novel Kruppel type zinc finger protein, represses the human T cell leukemia virus type I long terminal repeat-mediated expression. AB - We have shown that human T-cell leukemia virus type I (HTLV-I) gene expression is negatively regulated by the U5 repressive element (U5RE) of its long terminal repeat (LTR). To isolate factors binding to U5RE, we screened a cDNA expression library by south-western blotting with a U5RE probe. Screening 2 x10(6) clones gave a positive clone with a 3.8 kb insert encoding a novel 671 residue polypeptide, named HTLV-I U5RE binding protein 1 (HUB1), with five zinc finger domains and a Kruppel-associated box like domain; HUB1 may be related to a repressor belonging to the Kruppel type zinc finger protein. A 4.0 kb mRNA for HUB1 is ubiquitously expressed among all human tissues tested. HUB1 recognizes the TCCACCCC sequence as a core motif and exerts a strong repressive effect on HTLV-I LTR-mediated expression. A new repressive domain, named HUB1 repressive (HUR) domain, was identified, rather than the Kruppel-associated box like domain. The N-terminal region upstream of HUR domain seemed to be also indispensable to the repression. Thus, we propose that HUB1 is a new type repressor and plays an important role in the HTLV-I U5-mediated repression. PMID- 9396812 TI - Genetic interactions of conserved regions in the DEAD-box protein Prp28p. AB - The yeast PRP28 g ene has been implicated in nuclear precursor messenger RNA (pre mRNA) splicing, a two-step reaction involved in a multitude of RNA structural alterations. Prp28p, the gene product of PRP28 , is a member of the evolutionarily conserved DEAD-box proteins (DBPs). Members of DBPs are involved in a variety of RNA-related biochemical processes, presumably by their putative RNA helicase activities. Prp28p has been speculated to play a role in melting the duplex between U4 and U6 small nuclear RNAs (snRNAs), leading to the formation of an active spliceosome. To study the function of Prp28p and its interactions with other components of the splicing machinery, we have isolated and characterized a large number of prp28 conditional mutants. Strikingly, many of these prp28 mutations are localized in the highly conserved motifs found in all the DBPs. Intragenic reversion analysis suggests that regions of motifs II, III and V, as well as of motifs I and IV, in Prp28p are likely to be in close proximity to each other. Our results thus provide the first hint of the local structural arrangement for Prp28p, and perhaps for other DBPs as well. PMID- 9396813 TI - Identification of DNA replication and cell cycle proteins that interact with PCNA. AB - The identity of DNA replication proteins and cell cycle regulatory proteins which can be found in complexes involving PCNA were investigated by the use of PCNA immobilized on Sepharose 4B. A column containing bovine serum albumin (BSA) bound to Sepharose was used as a control. Fetal calf thymus extracts were chromatographed on PCNA-Sepharose and BSA-Sepharose. The columns were washed and then eluted with 0.5 M KCl. The salt eluates were examined for the presence of both DNA replication proteins (Pol alpha, delta, straightepsilon, PCNA, RFC, RFA, DNA ligase I, NDH II, Topo I and Topo II) and cell cycle proteins (Cyclins A, B1, D1, D2, D3, E, CDK2, CDK4, CDK5 and p21) by western blotting with specific antibodies. The DNA replication proteins which bound to PCNA-Sepharose included DNA polymerase delta and straightepsilon, PCNA, the 37 and 40 kDa subunits of RFC, the 70 kDa subunit of RPA, NDH II and topoisomerase I. No evidence for the binding of DNA polymerase alpha, DNA ligase I or topoisomerase II was obtained. Of the cell cycle proteins investigated, CDK2, CDK4 and CDK5 were bound. This study presents strong evidence that PCNA is a component of protein complexes containing DNA replication, repair and cell cycle regulatory proteins. PMID- 9396814 TI - Reactivation of denatured proteins by domain V of bacterial 23S rRNA. AB - In vitro transcripts containing domain V of the 23S rRNA of Escherichia coli and Bacillus subtilis can reactivate denatured proteins almost as efficiently as the total 23S rRNA. Here we show that almost the full length of domain V is required for reactivation of denatured pig muscle lactate dehydrogenase and pig heart cytoplasmic malate dehydrogenase: the central loop of this domain alone is not enough for this purpose. The antibiotic chloramphenicol, which binds to domain V of 23S rRNA, can inhibit reactivation of these proteins completely. Activity is eliminated by EDTA at a concentration of <1 mM, even in the presence of 4 mM MgCl2, suggesting that the three-dimensional conformation of the RNA should be maintained for this activity. PMID- 9396815 TI - The methylated DNA binding protein-2-H1 (MDBP-2-H1) consists of histone H1 subtypes which are truncated at the C-terminus. AB - The methylated DNA binding protein-2-H1 (MDBP-2-H1), present in rooster liver, is a member of the histone H1 family which inhibits transcription by binding selectively to methylated promoters. Here we have determined the primary structure of MDBP-2-H1. A comparison between histone H1 and MDBP-2-H1 was achieved by analyzing reversed phase HPLC-purified and V8-digested proteins by mass spectrometry and/or microsequencing. In rooster liver the most abundant histone H1 subtypes are H1 01 and H1 11L. Similarly, MDBP-2-H1 contains the same subtypes of histone H1. The histone H1 subtype H1 01 in MDBP-2-H1 has 150 amino acids, whereas the full-size histone H1 01 is 218 amino acids. The difference in mass between the two proteins is explained by C-terminal truncation of histone H1 01. PMID- 9396816 TI - Functional analysis of a replication origin from Saccharomyces cerevisiae: identification of a new replication enhancer. AB - Yeast replication origins have a modular arrangement of essential DNA sequences containing the ARS consensus sequence (ACS) flanked by auxiliary DNA elements which stimulate origin function. One of the auxiliary elements identified at several origins is a DNA replication enhancer that binds the Abf1p protein. We have isolated an ARS sequence from Saccharomyces cerevisiae based on its ability to bind Abf1p. Here we present a detailed molecular dissection of this ARS, designated ARS 1501, and we demonstrate that it functions as a genomic replication origin on chromosome XV . Mutagenesis of the Abf1p DNA-binding sites revealed that these sequences did not contribute significantly to ARS function. Instead, a new DNA element important for replication, designated REN1501, has been located 5' to the T-rich strand of the ACS. We show that REN1501 functions in either orientation and at variable distances from the ACS, defining this element as a DNA replication enhancer. Most significantly, point mutations within this element decreased the stability of plasmids bearing ARS 1501, suggesting that REN1501 binds a protein important for replication initiation. Only three elements found at origins are known to specifically bind proteins. These include the ARS essential sequences and the Abf1p and Rap1p DNA-binding sites. We show that the function of REN1501 at the origin cannot be replaced by a Rap1p DNA binding site or a site that binds the transcriptional factor Gal4p and can only be partially substituted for by an Abf1p recognition sequence. This implies that the role of the REN1501 element at the ARS 1501 origin is specific, and suggest that the frequency of origin firing in eukaryotic cells may be regulated by origin-specific enhancers. PMID- 9396817 TI - Nested genetic bit analysis (N-GBA) for mutation detection in the p53 tumor suppressor gene. AB - There is a growing and significant demand for reliable, simple and sensitive methods for repeated scanning of a given gene or gene fragment for detection and characterization of mutations. Solid-phase sequencing by single base primer extension of nested GBATM primers on miniaturized DNA arrays can be used to effectively scan targeted sequences for missense, insertion and deletion mutations. This paper describes the use of N-GBA arrays designed to scan the sequence of a 33 base region of exon 8 of the p53 gene (codons 272-282) encompassing a hot spot for mutations associated with the development of cancer. Synthetic DNA templates containing various missense, insertion and deletion mutations, as well as DNA prepared from pancreatic and biliary tumor cells, were genotyped using the exon 8 arrays. PMID- 9396819 TI - Minor groove DNA alkylation directed by major groove triplex forming oligodeoxyribonucleotides. AB - We describe sequence-specific alkylation in the minor groove of double-stranded DNA by a hybridization-triggered reactive group conjugated to a triplex forming oligodeoxyribonucleotide (TFO) that binds in the major groove. The 24 nt TFOs (G/A motif) were designed to form triplexes with a homopurine tract within a 65 bp target duplex. They were conjugated to an N 5-methyl-cyclopropapyrroloindole (MCPI) residue, a structural analog of cyclopropapyrroloindole (CPI), the reactive subunit of the potent antibiotic CC-1065. These moieties react in the DNA minor groove, alkylating adenines at their N3 position. In order to optimize alkylation efficiency, linkers between the TFO and the MCPI were varied both in length and composition. Quantitative alkylation of target DNA was achieved when the dihydropyrroloindole (DPI) subunit of CC-1065 was incorporated between an octa(propylene phosphate) linker and MCPI. The required long linker traversed one strand of the target duplex from the major groove-bound TFO to deliver the reactive group to the minor groove. Alkylation was directed by relative positioning of the TFOs. Sites in the minor groove within 4-8 nt from the end of the TFO bearing the reactive group were selectively alkylated. PMID- 9396818 TI - Test of the potential of a dATP surrogate for sequencing via MALDI-MS. AB - 1-(2'-Deoxy-beta-d-ribofuranosyl)-3-nitropyrrole phosphate was incorporated into a DNA decamer and analyzed via matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS). The extent and composition of the various fragment peaks were compared with those in the MALDI-MS spectrum of dT4AT5. The nitropyrrole containing oligomer proved to be more robust. Two different DNA template assays were then used to attempt to identify DNA replicating enzymes that would incorporate the corresponding triphosphate, i.e. 1-(2'-deoxy-beta-d ribofuranosyl)-3-nitropyrrole triphosphate (dXTP). It was shown that dXTP was not incorporated by some enzymes and it inhibited others. However, DNA polymerase I Klenow fragment and avian myeloblastosis virus reverse transcriptase incorporated dXTP in place of dATP and then replicated the template overhang in the usual way. The potential of dXTP as a surrogate for dATP in DNA sequencing with MALDI-MS analysis is discussed. PMID- 9396820 TI - Use of an engineered ribozyme to produce a circular human exon. AB - We report the use of an engineered ribozyme to produce a circular human exon in vitro. Specifically, we have designed a derivative of a yeast self-splicing group II intron that is able to catalyze the formation of a circular exon encoding the first kringle domain (K1) of the human tissue plasminogen activator protein. We show that the circular K1 exon is formed with high fidelity in vitro. Furthermore, the system is designed such that the circular exon that is produced consists entirely of human exon sequence. Thus, our results demonstrate that all yeast exon sequences are dispensable for group II intron catalyzed inverse splicing. This is the first demonstration that an engineered ribozyme can be used to create a circular exon containing only human sequences, linked together at a precise desired ligation point. We expect these results to be generalizable, so that similar ribozymes can be designed to precisely create circular derivatives of any nucleotide sequence. PMID- 9396821 TI - Retinoblastoma protein expression facilitates chromatin remodeling at the HLA-DRA promoter. AB - The major histocompatibility complex (MHC) class II genes encode a series of heterodimeric cell surface glycoproteins that bind peptide antigen. The MHC class II/peptide complex is bound by the T-cell receptor of CD4(+) T cells, thereby stimulating an immune response. The MHC class II genes are coordinately regulated by conserved promoter elements and are inducible by IFN-gamma. Furthermore, IFN gamma induction of the MHC class II genes in solid human tumor lines requires retinoblastoma protein (Rb). In vivo footprinting analyses of the HLA-DRA gene, which encodes the heavy chain subunit of the human MHC class II molecule, HLA-DR, revealed that Rb facilitates occupancy of multiple HLA-DRA promoter elements. Detecting the effect of Rb on HLA-DRA promoter occupancy in vivo required IFN gamma treatment. However, use of a variation on the in vivo footprinting technique, nuclei footprinting, which assays for promoter occupancy in isolated nuclei, revealed that expression of Rb facilitates promoter occupancy even in the absence of IFN-gamma. These results indicate that expression of Rb leads to modification of the chromatin environment of the HLA-DRA promoter independently of transcription. PMID- 9396822 TI - In vivo analyses of RNA polymerase I termination in Schizosaccharomyces pombe. AB - Recent studies on the termination of rDNA transcription by RNA polymerase I in Saccharomyces cerevisiae and Schizosaccharomyces pombe have suggested a more complex mechanism then previously described in higher eukaryotes. Termination appears to occur when a DNA-bound Reb1 protein molecule induces polymerase to pause in the context of a release element [see Reeder,R.H. and Lang,W. (1994) Mol. Microbiol ., 12, 11-15]. Because these conclusions in yeast were based entirely on in vitro analyses, we have examined the same termination process in S.pombe by expressing targeted mutations in vivo . S1nuclease protection studies indicate three tandemly arranged termination sites with most transcripts very efficiently terminated at the first site, 267 nt after the 3' end of the mature 25S rRNA sequence. Termination at each site is mediated by conserved terminator elements which bear limited sequence homology with that of mouse and also can be identified in S.cerevisiae . Removal of the first terminator element transfers dominance to the second site and construction of a new single terminator element at +150 still results in efficient termination and rRNA processing without a need for an additional upstream element. Genomic 'footprint' analyses and gel retardation assays confirm a process mediated by a strongly interacting protein factor but implicate an alternate binding site. Removal of the 5' flanking sequence or structure also had no effect on the site or efficiency of termination. Taken together the results in vivo suggest that the termination process in this fission yeast more strongly resembles the single element-mediated mechanism initially reported in mouse and is not dependent on additional upstream sequence as first reported in S.cerevisiae and postulated to function in general. PMID- 9396824 TI - Single substitutions of phosphorothioates in the HDV ribozyme G73 define regions necessary for optimal self-cleaving activity. AB - Phosphorothioate (NTPalphaS) analogues were incorporated into the HDV genomic ribozyme by transcription with T7 polymerase. The introduction of a sulfur in place of the pro-Rp oxygen at the phosphate 5'to positions A64, A63, A43, U27, G62, C61, C44, C41, C22and C21appeared to inhibit self-cleavage activity of the G73 genomic ribozyme. Except for position C22, elevated levels of Mg2+rescued the reaction to various extents. When the sites were identified in the RNA sequence, they were clustered in three distinct regions that, in the secondary structure models, are predicted to be primarily single-stranded. Two of these regions have been proposed to form extensive interactions that are thought to involve a homopurine base pair. The third region is thought to be directly associated with assembly of the cleavage site. PMID- 9396825 TI - Detection of a single base exchange in PCR-amplified DNA fragments using agarose gel electrophoresis containing bisbenzimide-PEG. AB - Using PCR fragments of known sequences derived from isolates of two related fungal species, simple submarine electrophoresis in agarose gels containing a bisbenzimide-PEG conjugate (H.A.-Yellow) has been shown to be capable of distinguishing DNA fragments 567 bp long which differ by as little as a single base change. However, only changes affecting bisbenzimide binding sites (which consist of at least four consecutive A/T bases) alter mobility; other changes are ineffective. A second ligand (H.A.-Red) with high G/C specificity is suggested which may be as effective in detecting other sequence changes. PMID- 9396823 TI - The proofreading domain of Escherichia coli DNA polymerase I and other DNA and/or RNA exonuclease domains. AB - Prior sequence analysis studies have suggested that bacterial ribonuclease (RNase) Ds comprise a complete domain that is found also in Homo sapiens polymyositis-scleroderma overlap syndrome 100 kDa autoantigen and Werner syndrome protein. This RNase D 3'-->5' exoribonuclease domain was predicted to have a structure and mechanism of action similar to the 3'-->5' exodeoxyibonuclease (proofreading) domain of DNA polymerases. Here, hidden Markov model (HMM) and phylogenetic studies have been used to identify and characterise other sequences that may possess this exonuclease domain. Results indicate that it is also present in the RNase T family; Borrelia burgdorferi P93 protein, an immunodominant antigen in Lyme disease; bacteriophage T4 dexA and Escherichia coli exonuclease I, processive 3'-->5' exodeoxyribonucleases that degrade single stranded DNA; Bacillus subtilis dinG, a probable helicase involved in DNA repair and possibly replication, and peptide synthase 1; Saccharomyces cerevisiae Pab1p dependent poly(A) nuclease PAN2 subunit, required for shortening mRNA poly(A) tails; Caenorhabditis elegans and Mus musculus CAF1, transcription factor CCR4 associated factor 1; Xenopus laevis XPMC2, prevention of mitotic catastrophe in fission yeast; Drosophila melanogaster egalitarian, oocyte specification and axis determination, and exuperantia, establishment of oocyte polarity; H.sapiens HEM45, expressed in tumour cell lines and uterus and regulated by oestrogen; and 31 open reading frames including one in Methanococcus jannaschii . Examination of a multiple sequence alignment and two three-dimensional structures of proofreading domains has allowed definition of the core sequence, structural and functional elements of this exonuclease domain. PMID- 9396826 TI - Microwave-assisted rapid deprotection of oligodeoxyribonucleotides. AB - A novel method for the deprotection of oligodeoxyribonucleotides under microwave irradiation has been developed. The oligodeoxynucleotides having base labile, phenoxyacetyl (pac), protection for exocyclic amino functions were fully deprotected in 0. 2 M sodium hydroxide (methanol:water : : 1:1, v/v) = A and 1 M sodium hydroxide (methanol:water : : 1:1, v/v) = B using microwaves in 4 and 2 min, respectively. The deprotection of oligodeoxyribonucleotides carrying conventional protecting groups, dAbz, dCbzand dGpac, for exocyclic amino functions was achieved in 4 min in B without any side product formation. The deprotected oligonucleotides were compared with the oligomers deprotected using standard deprotection conditions (29% aq. ammonia, 16 h, 55 degrees C) with respect to their retention time on HPLC and biological activity. PMID- 9396827 TI - RAPD-based screening of genomic libraries for positional cloning. AB - RAPD markers are frequently used for positional cloning. However, RAPD markers often contain repeated sequences which prevent genomic library screening by hybridisation. We have developed a simple RAPD analysis of genomic libraries based on the identification of cosmid pools and clones amplifying the RAPD marker of interest. Our method does not require the cloning or characterisation of the RAPD marker as it relies on the analysis of cosmid pools or clones using a simple RAPD protocol. We applied this strategy using four RAPD markers composed of single copy or repeated sequences linked to avirulence genes of the rice blast fungus Magnaporthe grisea . Cosmids containing these RAPD markers were easily and rapidly identified allowing the construction of physical contigs at these loci. PMID- 9396828 TI - Antibody-ribosome-mRNA (ARM) complexes as efficient selection particles for in vitro display and evolution of antibody combining sites. AB - We describe a rapid, eukaryotic, in vitro method for selection and evolution of antibody combining sites using antibody-ribosome-mRNA (ARM) complexes as selection particles. ARMs carrying single-chain (VH/K) binding fragments specific for progesterone were selected using antigen-coupled magnetic beads; selection simultaneously captured the genetic information as mRNA, making it possible to generate and amplify cDNA by single-step RT-PCR on the ribosome-bound mRNA for further manipulation. Using mutant libraries, antigen-binding ARMs were enriched by a factor of 10(4)-10(5)-fold in a single cycle, with further enrichment in repeated cycles. While demonstrated here for antibodies, the method has the potential to be applied equally for selection of receptors or peptides from libraries. PMID- 9396829 TI - A 71-kDa protein from Halobacterium salinarium belongs to a ubiquitous P-loop ATPase superfamily with head-rod-tail structure. AB - The nucleotide sequence of a genomic fragment from Halobacterium salinarium containing an open reading frame encoding a protein with a calculated molecular mass of 71 kDa was determined. Database searches revealed that this protein, Hp71, has similarities to eukaryotic cytoskeletal proteins. Heterologous production of Hp71 in Escherichia coli allowed the isolation of anti-Hp71 antibodies. The antibodies were used (1) to verify the production of Hp71 in H. salinarium and (2) to determine its cytoplasmic localization by immune electron microscopy. Homologous overproduction of Hp71 in H. salinarium and heterologous production in Haloferax volcanii resulted in modifications of cell morphology from rods to extended rods, and from pleiomorphic cells to rods, respectively. Structure prediction methods indicated that Hp71 has a head-rod-tail configuration, including an N-terminal domain with a nucleotide binding motif (P loop), and an extended discontinuous coiled-coil domain of 330 amino acids. To identify related proteins, the complete genomes of Haemophilus influenzae, Mycoplasma genitalium, and Methanococcus jannaschii were searched for deduced proteins with extended coiled-coil domains. Only one or two proteins were found for each organism, showing that Hp71 is one of only a few prokaryotic intracellular proteins with extended coiled-coil domains. The phenotype upon overproduction and the similarity of Hp71 to the SMC superfamily of P-loop head rod-tail proteins (named after SMC1, which is involved in the "stability of minichromosomes" in yeast) indicate that Hp71 might be involved in cytoskeleton formation and/or chromosome partitioning in H. salinarium. PMID- 9396830 TI - Growth at low temperature causes nitrogen limitation in the cyanobacterium Synechococcus sp. PCC 7002. AB - The coloration of cells of the cyanobacterium Synechococcus sp. PCC 7002 changed from normal blue-green to yellow-green when cells were grown at 15 degrees C in a medium containing nitrate as the sole nitrogen source. This change of coloration was similar to a general response to nutrient deprivation (chlorosis). For the chlorotic cells at 15 degrees C, the total amounts of phycobiliproteins and chlorophyll a decreased, high levels of glycogen accumulated, and growth was arithmetic rather than exponential. These changes in composition and growth occurred in cells grown at low (50 microE m-2 s-1) as well as high (250 microE m 2 s-1) light intensity. After a temperature shift-up to 38 degrees C, chlorotic cells rapidly regained their normal blue-green coloration and normal exponential growth rate within 7 h. When cells were grown at 15 degrees C in a medium containing urea as the reduced nitrogen source, cells grew exponentially and the symptoms of chlorosis were not observed. The decrease in photosynthetic oxygen evolution activity at low temperature was much smaller than the decrease in growth rate for cells grown on nitrate as the nitrogen source. These studies demonstrate that low-temperature-induced chlorosis of Synechococcus sp. PCC 7002 is caused by nitrogen limitation and is not the result of limited photosynthetic activity or photodamage to the photosynthetic apparatus, and that nitrogen assimilation is an important aspect of the low-temperature physiology of cyanobacteria. PMID- 9396832 TI - Purification and characterization of 9-hexadecenoic acid cis-trans isomerase from pseudomonas sp. strain E-3 AB - A 9-hexadecenoic acid cis-trans isomerase (9-isomerase) that catalyzed the cis-to trans isomerization of the double bond of free 9-cis-hexadecenoic acid [16:1(9c)] was purified to homogeneity from an extract of Pseudomonas sp. strain E-3 and characterized. Electrophoresis of the purified enzyme on both incompletely denaturing and denaturing polyacrylamide gels yielded a single band of a protein with a molecular mass of 80 kDa, suggesting that the isomerase is a monomeric protein of 80 kDa. The 9-isomerase, assayed with 16:1(9c) as a substrate, had a specific activity of 22.8 &mgr;mol h-1 (mg protein)-1 and a Km of 117.6 mM. The optimal pH and temperature for catalysis were approximately pH 7-8 and 30 degrees C, respectively. The 9-isomerase catalyzed the cis-to-trans conversion of a double bond at positions 9, 10, or 11, but not that of a double bond at position 6 or 7 of cis-mono-unsaturated fatty acids with carbon chain lengths of 14, 15, 16, and 17. Octadecenoic acids with a double bond at position 9 or 11 were not susceptible to isomerization. These results suggest that 9-isomerase has a strict specificity for both the position of the double bond and the chain length of the fatty acid. The enzyme catalyzed the cis-to-trans isomerization of fatty acids in a free form, and in the presence of a membrane fraction it was also able to isomerize 16:1(9c) esterified to phosphatidylethanolamine. The 9-isomerase was strongly inhibited by catecholic antioxidants such as alpha-tocopherol and nordihydroguaiaretic acid, but was not inhibited by 1, 10-phenanthroline or EDTA or under anoxic conditions. Based on these results, the possible mechanism of catalysis by this enzyme is discussed. PMID- 9396831 TI - Alteration of low-temperature susceptibility of the cyanobacterium Synechococcus sp. PCC 7002 by genetic manipulation of membrane lipid unsaturation. AB - Cyanobacteria acclimate to low temperature by desaturating their membrane lipids. Mutant strains of Synechococcus sp. PCC 7002 containing insertionally inactivated desA (Delta12 acyl-lipid desaturase) and desB (omega3 acyl-lipid desaturase) genes were produced, and their low-temperature susceptibility was characterized. The desA mutant synthesized no linoleic acid or alpha-linolenic acid, and the desB mutant did not produce alpha-linolenic acid. The desA mutant grew more slowly than the wild-type at 22 degrees C and could not grow at 15 degrees C. The desB mutant could not continuously grow at 15 degrees C, although no observable phenotype appeared at higher temperatures. It has been shown that expression of the desA gene occurs at 38 degrees C and is up-regulated at 22 degrees C, and that the desB gene is only expressed at 22 degrees C. These results indicate that the expression of the desA and desB genes occurs at higher temperatures than those at which a significant decline in physiological activities is caused by the absence of their products. The temperature dependency of photosynthesis was not affected by these mutations. Since chlorosis and inability to grow at 15 degrees C with nitrate was suppressed by the substitution of urea as a nitrogen source, it is very likely that the chilling susceptibility of the desaturase mutants is attributable to nutrient limitation. PMID- 9396833 TI - Degradation of p-nitrophenol by the phototrophic bacterium Rhodobacter capsulatus. AB - The phototrophic bacterium Rhodobacter capsulatus detoxified p-nitrophenol and 4 nitrocatechol. The bacterium tolerated moderate concentrations of p-nitrophenol (up to 0.5 mM) and degraded it under light at an optimal O2 pressure of 20 kPa. The bacterium did not metabolize the xenobiotic in the dark or under strictly anoxic conditions or high O2 pressure. Bacterial growth with acetate in the presence of p-nitrophenol took place with the simultaneous release of nonstoichiometric amounts of 4-nitrocatechol, which can also be degraded by the bacterium. Crude extracts from R. capsulatus produced 4-nitrocatechol from p nitrophenol upon the addition of NAD(P)H, although at a very low rate. A constitutive catechol 1, 2-dioxygenase activity yielding cis,cis-muconate was also detected in crude extracts of R. capsulatus. Further degradation of 4 nitrocatechol included both nitrite- and CO2-releasing steps since: (1) a strain of R. capsulatus (B10) unable to assimilate nitrate and nitrite released nitrite into the medium when grown with p-nitrophenol or 4-nitrocatechol, and the nitrite concentration was stoichiometric with the 4-nitrocatechol degraded, and (2) cultures of R. capsulatus growing microaerobically produced low amounts of 14CO2 from radiolabeled p-nitrophenol. The radioactivity was also incorporated into cellular compounds from cells grown with uniformly labeled 14C-p-nitrophenol. From these results we concluded that the xenobiotic is used as a carbon source by R. capsulatus, but that only the strain able to assimilate nitrite (E1F1) can use p-nitrophenol as a nitrogen source. PMID- 9396834 TI - Skew or third moment of bacterial generation times. AB - We studied two statistical hypotheses for the occurrence of cellular division and compared these hypotheses to available data. The two models were tested by observed distributions of cellular size during steady-state growth. The 30-year old sloppy size model could be rejected, whereas the recently developed incremental size proposal could not. The latter proposition was accepted by default. We concluded that the time between successive divisions is not simply derived from extant size at cellular division, but rather from interdivisional size increment. We therefore propose that cellular division is regulated by the need of cells at birth to accumulate a certain amount of mass or something related to mass before division. PMID- 9396835 TI - The Alcaligenes eutrophus hemN gene encoding the oxygen-independent coproporphyrinogen III oxidase, is required for heme biosynthesis during anaerobic growth. AB - The insertion mutant HF231 of Alcaligenes eutrophus H16 failed to grow anaerobically on nitrate and nitrite. When grown under oxygen limitation, mutant HF231 specifically excreted coproporphyrin III, an intermediate of heme biosynthesis. With the help of a Tn5-labeled fragment, we identified and cloned the corresponding wild-type fragment. Sequence analysis of the mutant locus revealed an open reading frame consisting of 1,473 bp, predicting a protein of 491 amino acids that corresponds to a size of 54.2 kDa. In the non-coding upstream region, consensus elements that are indicative for binding sites of the anaerobic transcriptional regulator Fnr were identified. The deduced polypeptide showed extensive sequence similarity with various bacterial oxygen-independent coproporphyrinogen III oxidases designated HemN. HemN catalyzes the oxidative decarboxylation of coproporphyrinogen III to yield protoporphyrinogen IX. Anaerobic growth on nitrate and nitrite of mutant HF231 was restored by introducing the hemN gene of A. eutrophus or of Pseudomonas aeruginosa on a broad host-range vector. Likewise, the A. eutrophus hemN complemented heme biosynthesis of a Salmonella typhimurium hemF/hemN double mutant during anaerobic and aerobic growth. Analysis of a transcriptional lacZ gene fusion showed that expression of hemN in A. eutrophus is nitrate-independent and repressed by oxygen. PMID- 9396836 TI - Analysis of changes in congener selectivity during PCB degradation by Burkholderia sp. strain TSN101 with increasing concentrations of PCB and characterization of the bphBCD genes and gene products. AB - We isolated and characterized a gram-negative bacterium, Burkholderia sp. strain TSN101, that can degrade polychlorinated biphenyls (PCBs) at concentrations as high as 150 microg Kaneclor 300/ml, a PCB mixture equivalent to Aroclor 1242. Growing cells of strain TSN101 degraded most of the tri- and tetrachlorobiphenyls in medium containing 25 microg Kaneclor 300/ml. Using PCB concentrations of 50 150 microg of Kaneclor 300/ml, the congener selectivity pattern was different and the pattern of chlorine substitution strongly affected degradation of some congeners. At 25 microg Kaneclor 300/ml, strain TSN101 degraded di- and trichlorinated congeners with chlorine substitutions at both the ortho and the para positions. At higher concentrations of Kaneclor 300, di- and trichlorobiphenyls with ortho substituents in both phenyl rings were not degraded well. Trichlorobiphenyls with para and meta substitutents were degraded equally well at all concentrations studied. The ability of strain TSN101 to degrade ortho and para-substituted congeners was confirmed using a defined PCB mixture with chlorine substituents at 2'- and 4'-positions. A 5-kb DNA fragment containing the bphBCD genes was cloned and sequenced. Comparison of the deduced amino acid sequences of these genes with related proteins indicated 99 and 98% sequence similarity to the BphB and BphD of Comamonas testosteroni strain B-356, respectively. The bphC gene product showed 74% sequence similarity to the BphC of Burkholderia cepacia strain LB400 and exhibited a narrow substrate specificity with strong affinity for 2, 3-dihydroxybiphenyl. A bphC-disrupted mutant of Burkholderia sp. strain TSN101, constructed by gene replacement, lost the ability to utilize biphenyl, thus supporting the role of the cloned bph gene in biphenyl metabolism. PMID- 9396838 TI - Amino acid degradation by the mesophilic sulfate-reducing bacterium desulfobacterium vacuolatum AB - Desulfobacterium vacuolatum strain IbRM was able to grow using casamino acids as a source of carbon, energy and nitrogen. Growth was accompanied by utilization of several amino acids and sulfide production. Proline and glutamate were used preferentially and to the greatest extent. Glycine, serine and alanine were used more slowly and only after proline and glutamate were used. Isoleucine, valine, leucine and aspartate decrease was slowest and occurred in a linear fashion throughout the growth phase. Amino acids used from casamino acids, excluding aspartate, were also used as single carbon, energy and nitrogen sources. As a single amino acid, aspartate could only be used as a nitrogen source. Aspartate was not used as an electron acceptor. No growth occurred on any amino acid in the absence of sulfate. As single substrates, isoleucine, proline and glutamate were oxidized without formation of acetate and with molar yields of 13.1, 9.4 and 7.7 g mol-1, respectively. PMID- 9396837 TI - Functional expression in Escherichia coli of the Haemophilus influenzae gene coding for selenocysteine-containing selenophosphate synthetase. AB - The selenophosphate synthetases from several organisms contain a selenocysteine residue in their active site where the Escherichia coli enzyme contains a cysteine. The synthesis of these enzymes, therefore, depends on their own reaction product. To analyse how this self-dependence is correlated with the selenium status, e.g. after recovery from severe selenium starvation, we expressed the gene for the selenocysteine-containing selenophosphate synthetase from Haemophilus influenzae (selDHI) in an E. coli DeltaselD strain. Gene selDHI gave rise to a selenium-containing gene product and also supported - via its activity - the formation of E. coli selenoproteins. The results provide evidence either for the suppression of the UGASec codon with the insertion of an amino acid allowing the formation of a functional product or for a bypass of the selenophosphate requirement. We also show that the selenocysteine synthesis and the insertion systems of the two organisms are fully compatible despite conspicuous differences in the mRNA recognition motif. PMID- 9396839 TI - Spontaneous mutation in a thermoacidophilic archaeon: evaluation of genetic and physiological factors AB - We used direct selection of pyrE and pyrF mutants to estimate the rates of spontaneous mutation in Sulfolobus acidocaldarius as a function of genetic background and culture conditions. Fluctuation tests were applied to several genetically marked strains, including one isolated as a putative mutator strain, and to cultures grown over a wide range of temperature and other physiological conditions. The results suggested some impact of auxotrophic markers on the apparent rate of mutation, but no obvious pattern of effect of growth conditions, including those that gave evidence of being physiologically stressful. PMID- 9396840 TI - Tetrahydrofolate serves as a methyl acceptor in the demethylation of dimethylsulfoniopropionate in cell extracts of sulfate-reducing bacteria. AB - Tetrahydrofolate was shown to function as a methyl acceptor in the anaerobic demethylation of dimethylsulfoniopropionate to methylthiopropionate in cell extracts of the sulfate-reducing bacterium strain WN. Dimethylsulfoniopropionate dependent activities were 0.56 micromol methyltetrahydrofolate min-1 (mg protein) 1 and were higher than required to explain the growth rate of strain WN on dimethylsulfoniopropionate. The reaction did not require ATP or reductive activation by titanium(III)-nitrilotriacetic acid. Preincubation of the extract under air significantly decreased the activity (35% loss in 3 h). Three other dimethylsulfoniopropionate-demethylating sulfate reducers, Desulfobacterium niacini, Desulfobacterium vacuolatum, and Desulfobacterium strain PM4, had dimethylsulfoniopropionate:tetrahydrofolate methyltransferase activities of 0.16, 0.05, and 0.24 micromol min-1 (mg protein)-1, respectively. No methyltransferase activity to tetrahydrofolate was found with betaine as a substrate, not even in extracts of betaine-grown cells of these sulfate reducers. Dimethylsulfoniopropionate demethylation in cell extracts of strain WN was completely inhibited by 0.5 mM propyl iodide; in the light, the inhibition was far less strong, indicating involvement of a corrinoid-dependent methyltransferase. PMID- 9396841 TI - Desulfovibrio inopinatus, sp. nov., A new sulfate-reducing bacterium that degrades hydroxyhydroquinone (1,2,4-trihydroxybenzene) PMID- 9396842 TI - Rashkind dedication. PMID- 9396843 TI - History of pediatric interventional catheterization: pediatric therapeutic cardiac catheterizations. PMID- 9396844 TI - "Physicians must abandon the illusion of autonomy . . .". AB - Radical changes are occurring in medicine which are impacting the practicing physician. This is especially true for specialists caring for very acutely ill patients such as those with congenital heart disease. This article explores developments such as the reversal of fiscal and medical paradigms, risk-averse behavior, price-based costing, and competing physician priorities. Special emphasis is directed to issues in pediatric cardiology. Suggestions for action are made, the primary of which is that physicians must abandon the illusion of autonomy in order to acquire the reality of collective influence on the practice of medicine. PMID- 9396845 TI - Balloon pulmonary valvuloplasty. PMID- 9396846 TI - Innovative approaches to interventional pediatric cardiology: different pokes for different folks. PMID- 9396847 TI - Transcatheter treatment of coarctation of the aorta: a review. PMID- 9396848 TI - Catheterization treatment of stenosis and hypoplasia of pulmonary arteries. AB - Stenosis of pulmonary arteries is one of the most challenging problems requiring treatment in the care of patients with congenital and acquired cardiopulmonary disease. Surgical approaches have been met with difficulty over the years, and may themselves lead to further distortion of the treated arteries. Balloon dilation first came into use in the 1980s, and has proved moderately effective. Its use has been extended to proximal pulmonary valve stenosis in order to improve distal flow and artery growth in some variants of tetralogy of Fallot. More recently, the judicious application of stent implantation has improved the outlook for pulmonary artery stenosis. The etiology, treatment (with balloon dilation and stent placement), and prognosis of pulmonary arterial stenosis will be discussed. PMID- 9396849 TI - Transcatheter management of venous stenosis. PMID- 9396850 TI - Oops-the balloon burst before the stent was deployed! PMID- 9396852 TI - Pediatric cardiovascular embolization therapy. AB - Transcatheter embolization of superfluous vascular structures has assumed an important role in pediatric interventional cardiology. A variety of devices and materials are being used to treat an increasing number of unwanted arterial, venous, and surgically created vascular connections. In general, the occlusion techniques are simple, the results are good, and the complication rates are low. The current indications, devices, materials, methods, applications, and results of pediatric cardiovascular embolization therapy are described. PMID- 9396851 TI - Transcatheter management of patent ductus arteriosus. PMID- 9396853 TI - Per-catheter ASD closure. AB - Per-catheter devices for atrial septal defect (ASD) closure have been evolving since 1974. The four major devices available for use on a limited basis in early 1997 are reviewed. These include (in alphabetical order) the Angel Wing device, the ASDOS device, the Buttoned device, and the CardioSeal device (successor to the Clamshell). Sufficient data have been collected to indicate that transcatheter ASD closure is a viable alternative to surgery in selected patients. The advantages of the concept of per-catheter closure over surgical closure should lead to the continued development of devices and techniques for per-catheter treatment of ASD and other septal defects in the years to come. PMID- 9396854 TI - Electrical/ablational therapeutic cardiac catheterization. PMID- 9396855 TI - Interventional pediatric cardiology: state of the art and future directions. AB - Although the interventional pediatric cardiology began in the early 1950s, it was not until the mid-1980s that a full spectrum of transcatheter interventions in children could be undertaken including balloon atrial septostomy which has been in usage since 1966. Enormous developments have occurred even from the mid-1980s to date. In this review, current state-of-the-art for each broad area of therapeutic catheterization is presented. A large variety of lesions could be opened-up or closed, as the case may be and the results of these interventions were either similar to or better than those reported for the alternative surgical therapy. Indeed, therapeutic catheterization techniques have replaced the conventional surgery for many lesions and are threatening to do so for others. However, long-term follow-up results are scanty and are needed. Further miniaturization of catheters/sheaths used in interventional pediatric cardiology and development of new technology for the lesions which are not amenable to currently available transcatheter methods are awaited. The future seems to be bright for interventional pediatric cardiology. PMID- 9396856 TI - Molecular analysis of skewed Tcra-V gene use in T-cell receptor beta-chain transgenic mice. AB - The influence of beta-chain diversity on the expressed T-cell receptor (TCR) alpha-chain repertoire was investigated using transgenic mice which exclusively express a single rearranged TCR beta-chain gene. Analysis of these mice using alpha-chain-specific recombinant cDNA libraries showed that expression of the transgene-encoded beta chain results in significant skewing in Tcra-V gene segment usage vs nontransgenic mice. Skewing was most pronounced towards alpha chains using TCRA-V segments. Sequence analysis of Tcra-V8-containing genes from transgenic T cells revealed predominant use of a single Tcra-J segment (Tcra J24), which was not detected in Tcra-V8 containing genes isolated from nontransgenic T cells. Further analysis revealed that co-expression of Tcra-V8 with Tcra-J24 in beta-transgenic mice is exhibited almost exclusively by CD4+ T cells, and is associated with a limited number of closely related N-regions. Analysis of transgenic CD8+ T cells demonstrated predominant co-expression of Tcra-V8 with another Tcra-J (Tcra-J30), together with a different, limited N region sequence. We conclude that the composition of expressed beta chains can profoundly influence the selection of companion alpha chains expressed in the periphery, and that alpha-chain N and J regions play a crucial role in discriminating between class I vs class II major histocompatibility complex (MHC) restricted recognition. Further, these results are in agreement with recent data concerning the crystal structure of the TCR, and most consistent with a model for TCR structure in which the complementarity determining region (CDR)3alpha domain participates in direct contact with the MHC. PMID- 9396857 TI - Characterization of chimpanzee TCRV gene polymorphism: how old are human TCRV alleles? AB - The functional relevance of the majority of human T-cell receptor A and B variable region gene polymorphisms is controversial. Studies of human and nonhuman primate major histocompatibility complex (MHC) class I and II polymorphisms show that allelic lineages predate human speciation and indicate that selection favors the long-term maintenance of these advantageous mutations. We investigated at the DNA level whether 15 human TCRA and B polymorphisms exist in contemporary chimpanzee populations. Polymorphisms consisted of variable region replacements, a recombination signal sequence base change, and silent mutations. With one exception, none of these human TCR polymorphisms were observed in contemporary chimpanzees. Investigation of the same polymorphisms in a range of other nonhuman primates showed little evidence of the existence of human polymorphism prespeciation. Chimpanzee TCRAV and BV regions were however polymorphic for variation so far not observed in human groups. Levels of mitochondrial and nuclear DNA sequence variation in contemporary chimpanzees suggest that population bottlenecks have not been a feature of chimpanzee evolution and it is therefore probable that most human TCR polymorphisms have evolved in the estimated five million years since the speciation of human and chimpanzees. Thus, over the evolutionary time period studied, ancient TCRA and B polymorphisms have not been maintained by selection to the same degree as postulated for MHC polymorphisms. PMID- 9396858 TI - T-cell receptor variable alpha (TCRAV) polymorphisms in European, Chinese, South American, AfroCaribbean, and Gambian populations. AB - Interactions involving the T-cell receptor (TCR) and major histocompatibility complex (MHC) are fundamental to the generation of a specific immune response. The study of interpopulation differences in TCR genes may identify those genes which are subject to selection, and also provides useful information for future genetic studies in these populations. In this study we present analysis of five TCRAV polymorphisms, for V5S1, V6S1, V8S1, V17S1, and V21S1 loci in five human populations by single-strand conformational polymorphism (SSCP) analysis. Caucasian, Chinese, Gambian, AfroCaribbean, and South American Indians (Mapuches) showed marked interpopulation variation for both the silent (V5S1, V17S1, and V21S1) and coding (V6S1 and V8S1) polymorphisms. In general the alleles were conserved in the different populations, but new, additional variants were found for V5S1 and V17S1 in Gambians and Caucasians. V6S1 overall showed the highest nucleotide diversity, and V6S1 genotype distributions were skewed away from expected values in Chinese and Mapuches. Analysis of allelic associations showed a general lack of linkage disequilibrium between the loci, which was reflected by the absence of strong population-specific haplotypes. PMID- 9396859 TI - Characterization of 12 microsatellite loci of the human MHC in a panel of reference cell lines. AB - The human genome contains a large number of interspersed microsatellite repeats which exhibit a high degree of polymorphism and are inherited in a Mendelian fashion, making them extremely useful genetic markers. Several microsatellites have been described in the HLA region, but allele nomenclature, a set of broadly distributed controls, and typing methods have not been standardized, which has resulted in discrepant microsatellite data between laboratories. In this report we present a detailed protocol for genotyping microsatellites using a semi automated fluorescence-based method. Twelve microsatellites within or near the major histocompatibility complex (MHC) were typed in the 10th International Histocompatibility Workshop homozygous typing cell lines (HTCs) and alleles were designated based on size. All loci were sequenced in two HTCs providing some information on the level of complexity of the repeat sequence. A comparison of allele size obtained by genotyping versus that obtained by direct sequencing showed minor discrepancies in some cases, but these were not unexpected given the technical differences in the methodologies. Fluorescence-based typing of microsatellites in the MHC described herein is highly efficient, accurate, and reproducible, and will allow comparison of results between laboratories. PMID- 9396860 TI - MICA, a new polymorphic HLA-related antigen, is expressed mainly by keratinocytes, endothelial cells, and monocytes. AB - MICA is a new polymorphic gene in the HLA region expressed in epithelial cell lines and gastrointestinal epithelium. Little is yet known about the MICA protein, and the pattern of its expression by freshly isolated cells has not been established. In the present experiments, we used antibodies raised in rabbits against alpha1 and alpha2 domain-peptides to study the expression of MICA. By western blot and immunoprecipitation, we detected a band of 62 000 Mr in various cell lines (THP-1, U937, HeLa, A431, Raji, MOLT-4, and HUV-EC-C) and in freshly isolated keratinocytes, endothelial cells, and monocytes but not in CD4+ and CD8+ T cells, and CD19+ cells (B lymphocytes). It was not possible to up-regulate the expression of MICA in different cells by stimulation with gamma-interferon, but the expression of MICA was induced in phytohemagglutinin-stimulated T cells. We confirmed that MICA is expressed at the cell surface by flow cytometry. Results of immunoprecipitation studies of beta2-microglobulin (beta2m)- or MICA-depleted, metabolically labeled HeLa cells indicated that MICA was not associated with beta2m. Although the function of MICA is still unknown, its restricted pattern of tissue expression, the fact that it is expressed on the cell surface, and its polymorphic nature suggest that this new molecule, encoded close to HLA class I, may play a role in the interaction between epithelial cells and cells of the immune system. PMID- 9396861 TI - Crucial role of N-terminal residue of binding peptides in recognition of the monoclonal antibody specific for the peptide-HLA-B5, -B35 complex. AB - The monoclonal antibody (mAb) 4D12 specific for the HLA-B5, -B35 cross-reacting group (CREG) bound to a fraction of HLA-B*3501 and HLA-B*5101 molecules carrying self-peptides. Analysis of the binding of mAb 4D12 to HLA-B*3501 and -B*5101 molecules pulsed with chemically synthesized peptides revealed that this mAb recognizes a restricted number of peptides and that P1 of the bound peptides critically influences its binding. The 4D12 mAb bound only to HLA-B*3501 molecules carrying peptides with Asn, Asp, Glu, Ser, and Val at P1. Analysis using an HLA-B*3501 crystallographic model suggested that 4D12 may recognize the side chain of the P1 residue that is pointing to the solvent. On the other hand, 4D12 bound only to HLA-B*5101 molecules carrying peptides with Asn or Asp at P1, suggesting that the 4D12 epitope formed by Glu, Ser, or Val at P1 and the A pocket was changed by the substitution of His for Tyr at residue 171 of HLA B*3501 molecules. This was confirmed by testing the binding of mAb 4D12 to HLA B*3501 mutant molecules at residue 171 carrying these peptides. These results together suggest that the conformation of the A-pocket and its hydrogen bound network with the P1 residue is also critical for the binding of mAb 4D12. The present study shows the molecular basis of the specificity of 4D12 for the peptide-HLA class I complex. PMID- 9396862 TI - Four dominant loci for the vascular responses by the antitumor polysaccharide, lentinan. AB - Lentinan, beta-1,6;1,3-glucan, showing an antitumor effect against mouse solid type tumors, can induce marked vascular dilation and hemorrhage (VDH) in very localized areas such as the ears, feet, and tails of mice in the early stages after its administration (Maeda et al. 1984). VDH has been found to be one of the T-cell-mediated responses triggered by lentinan. We reported previously that the responsiveness of mice to lentinan with respect to VDH induction is controlled by a dominant gene(s), Ltn2 (formerly), and that no sex difference was observed (Maeda et al. 1991). To determine the chromosomal location of the Ltn2 gene(s), we typed genomic DNAs of 193 N2 segregants of crosses between a high responder MA/MyJ and a low responder AKR/J by the polymerase chain reaction-simple sequence length polymorphism technique using 83 chromosome-specific microsatellite markers. We identified one major gene (Ltnr3) and three minor genes (Ltnr4, Ltnr5, and Ltnr6) responsible for the VDH induction. Ltnr3 was closely linked to D6Mit135 on chromosome 6 (P <0.00000) and Ltnr4, Ltnr5, and Ltnr6 to D9Mit161 on chromosome 9 (P <0.00032), D15Mit147 on chromosome 15 (P <0.00014) and D16Mit4 on chromosome 16 (P <0.00014), respectively. PMID- 9396863 TI - Cytogenetic orientation of the rat major histocompatibility complex (MHC) on chromosome 20. PMID- 9396864 TI - Molecular cloning of major histocompatibility complex class I cDNAs from the pufferfish Fugu rubripes. PMID- 9396865 TI - The mouse HFE gene. PMID- 9396866 TI - Conserved structure and tissue expression of rat eotaxin. PMID- 9396867 TI - The future of bone density measurements. PMID- 9396868 TI - Cardiofocal collimators for gated single-photon emission tomographic myocardial perfusion imaging. AB - Cardiofocal collimators (CFCs) are more sensitive than parallel-hole collimators (PHCs) of the same resolution because the rays converge in the centre of the field of view. After reconstruction a useful field of view with a 10 cm radius in which both sensitivity and resolution are homogeneous is obtained. In this article the feasibility and accuracy of gated single-photon emission tomographic (gated SPET) myocardial perfusion imaging using a triple-head camera equipped with CFC, is evaluated. Twenty patients with a history of myocardial infarction were studied. SPET myocardial perfusion images, gated in eight time bins, were acquired in a random sequence with a PHC and a CFC for each patient. Imaging was started 60 min after the injection of 925 MBq of technetium-99m tetrofosmin at rest. Ninety-six projections over 360 degrees were acquired, with 32 stops of 40 s for the PHC and 20 s for the CFC in order to obtain similar count densities. The extent (EXT) and severity (SEV) of perfusion defects were quantified on polar maps using the non-gated data. Left ventricular volumes [end-diastolic volume (EDV), end-systolic volume (ESV)] and ejection fraction (LVEF) were calculated from gated data using the Cedars-Sinai program. In 17 of 20 patients the complete left ventricle was positioned within the useful field of view of the CFC. The results in respect of perfusion, volumes and ejection fraction were almost identical to those obtained with the PHC. The mean difference+/-SD between the CFC and the PHC was -2.30+/-7.16 (% of LV area) for EXT, -0.48+/-2.90 for SEV (arbitrary units), -1,50+/-5.25 (ml) for EDV and 0.53+/-4.10 (%) for LVEF. The largest differences in EXT and LV volumes were observed in patients in whom a part of the LV was not positioned within the useful field of view. We conclude that, for the majority of patients, identical information with regard to both perfusion and function can be derived from gated SPET myocardial perfusion studies obtained with PHCs or with CFCs. Because of the greater sensitivity, however, a much shorter acquisition time is required with CFCs. PMID- 9396869 TI - Dual matrix ordered subsets reconstruction for accelerated 3D scatter compensation in single-photon emission tomography. AB - Three-dimensional (3D) iterative maximum likelihood expectation maximization (ML EM) algorithms for single-photon emission tomography (SPET) are capable of correcting image-degrading effects of non-uniform attenuation, distance-dependent camera response and patient shape-dependent scatter. However, the resulting improvements in quantitation, resolution and signal-to-noise ratio (SNR) are obtained at the cost of a huge computational burden. This paper presents a new acceleration method for ML-EM: dual matrix ordered subsets (DM-OS). DM-OS combines two acceleration methods: (a) different matrices for projection and back projection and (b) ordered subsets of projections. DM-OS was compared with ML-EM on simulated data and on physical thorax phantom data, for both 180 degrees and 360 degrees orbits. Contrast, normalized standard deviation and mean squared error were calculated for the digital phantom experiment. DM-OS resulted in similar image quality to ML-EM, even for speed-up factors of 200 compared to ML EM in the case of 120 projections. The thorax phantom data could be reconstructed 50 times faster (60 projections) using DM-OS with preservation of image quality. ML-EM and DM-OS with scatter compensation showed significant improvement of SNR compared to ML-EM without scatter compensation. Furthermore, inclusion of complex image formation models in the computer code is simplified in the case of DM-OS. It is thus shown that DM-OS is a fast and relatively simple algorithm for 3D iterative scatter compensation, with similar results to conventional ML-EM, for both 180 degrees and 360 degrees acquired data. PMID- 9396870 TI - Iodine-123 labeled nor-beta-CIT as a potential tracer for serotonin transporter imaging in the human brain with single-photon emission tomography. AB - Iodine-123 labelled 2beta-carbomethoxy-3beta-(4-iodophenyl) (nor-beta-CIT) is an analogue of beta-CIT, which has high affinity to the serotonin transporter. Initial single-photon emission tomography (SPET) studies with [123I]nor-beta-CIT were performed in five healthy volunteers. In addition, its metabolism in plasma was investigated with gradient high performance liquid chromatography. [123I]nor beta-CIT was prepared by a method which gave a specific radioactivity of more than 180 GBq/micromol. Unchanged [123I]nor-beta-CIT in plasma accounted for 43% and 19% of total radioactivity after 30 and 180 min, respectively. The dynamic SPET studies demonstrated a high and rapid uptake of radioactivity in the brain (6%/ID at 30 min). Highest accumulation was observed in the striatum, the mid brain and the thalamus. The specific binding in the mid-brain was 33% higher compared with that of [123I]beta-CIT. The high radioactivity in the mid-brain is assumed to represent the accumulation of [123I]nor-beta-CIT in the serotonin transporter-rich regions, which indicates that [123I]nor-beta-CIT might be a potential tracer for visualization of serotonin transporter sites in the human brain with SPET. PMID- 9396871 TI - Human biodistribution and dosimetry of [123I]FP-CIT: a potent radioligand for imaging of dopamine transporters. AB - This study reports on the biodistribution and radiation dosimetry of iodine-123 labelled N-omega-(flu- oropropyl)-2beta-carbomethoxy-3beta-(4-iodophenyl)tropane ([123I]FP-CIT), a promising radioligand for the imaging of dopamine transporters. In 12 healthy volunteers, conjugate whole-body scans were performed up to 48 h following intravenous injection of approximately 100 MBq [123I]FP-CIT. Attenuation correction was performed using a transmission whole-body scan obtained prior to injection of the radioligand, employing a 123I flood source. Blood samples were taken and urine was freely collected up to 48 h after injection of the radiotracer. For each subject, the percentage of injected activity measured in regions of interest over brain, striatum, lungs and liver were fitted to a multicompartmental model to give time-activity curves. The cumulative urine activity curve was used to model the urinary excretion rate and, indirectly, to predict faecal excretion. Using the MIRD method, nine source organs were considered in estimating absorbed radiation doses for organs of the body. The images showed rapid lung uptake and hepatobiliary excretion. Diffuse uptake and retention of activity was seen in the brain, especially in the striatum. At 48 h following the injection of [123I]FP-CIT, mean measured urine excretion was 60%+/-9% (SD), and mean predicted excretion in faeces was 14%+/-1%. In general, the striatum received the highest absorbed dose (average 0.23 mGy/MBq), followed by the urinary bladder wall (average 0.054 mGy/MBq) and lungs (average 0.043 mGy/MBq). The average effective dose equivalent of [123I]FP-CIT was estimated to be 0.024 mSv/MBq. The amount of [123I]FP-CIT required for adequate dopamine transporter imaging results in an acceptable effective dose equivalent to the patient. PMID- 9396872 TI - Pharmacological effects of dopaminergic drugs on in vivo binding of [99mTc]TRODAT 1 to the central dopamine transporters in rats. AB - The purpose of this study was to investigate the influence of drugs competing for the dopamine transporter (DAT) or changing intra- and/or extracellular dopamine levels on the binding of a novel technetium-99m labeled tropane derivative, technetium, [2-[[2-[[[3-(4-chloro- phenyl)-8-methyl-8-azabicyclo[3, 2, 1]oct-2 yl]methyl] (2-mercaptoethyl)amino]ethyl]amino]ethanethiolato(3)]-oxo-[1R-(exo exo)]-, [99mTc]TRODAT-1, to DAT. This paper describes the further characterization of [99mTc]TRODAT-1 binding sites in rats under conditions which may exist in patients receiving various drug treatments. All experiments were carried out using an i.v. injection of [99mTc]TRODAT-1 into male Sprague-Dawley rats. Measurements of % dose/gram ratio of (striatum-cerebellum)/cerebellum at 1 h post injection were used as an indicator for specific DAT binding. The biodistribution studies were performed in the presence of drugs which compete for the binding site, such as CFT (WIN 35,428) and methylphenidate, drugs which influence dopamine levels, such as L-DOPA, gamma-hydroxybutyrolactone, and alpha methyl-p-tyrosine, and d-amphetamine, which both acts as a competitor for DAT binding and increases dopamine levels. Additionally, the influence of dopamine receptor agonists, such as apomorphine and (+)bromocriptine, on biodistribution was tested. Binding of [99mTc]TRODAT-1 to DAT was found to be inhibited by CFT, methylphenidate, and d-amphetamine in a dose-dependent manner. The specific binding of [99mTc]TRODAT-1 was not altered by dopamine receptor agonists or by drugs which cause minor changes in dopamine levels. When administered in high doses (634 micromol/kg), L-DOPA also decreased the binding of [99mTc]TRODAT-1. It is likely that a low dose of L-DOPA (normally needed in the treatment of Parkinson's disease) will not affect the results on [99mTc]TRODAT-1 single-photon emission tomographic (SPET) imaging studies. In conclusion, the results clearly demonstrate the specificity of [99mTc]TRODAT-1 binding to DAT in vivo. Competition for [99mTc]TRODAT-1 binding was observed only with drug treatment that significantly increases dopamine levels or actively competes for binding at DAT. The results suggest that prior knowledge of whether patients are receiving various drug treatments may assist in the interpretation of DAT status as assessed by SPET imaging studies using [99mTc]TRODAT-1. PMID- 9396873 TI - Biodistribution and dosimetry of iodine-123-labelled Z-MIVE: an oestrogen receptor radioligand for breast cancer imaging. AB - This study reports on the distribution and radiation dosimetry of iodine-123 labelled cis-11beta-methoxy-17alpha-iodovinyloestradiol (Z-[123I]MIVE), a promising radioligand for imaging of oestrogen receptors (ERs) in human breast cancer. Whole-body scans were performed up to 24 h after intravenous injection of 138-193 MBq Z-[123I]MIVE in five healthy female volunteers, four with and one without thyroid blockade. Blood samples were taken at various times up to 24 h after injection. Urine was collected up to 24 h after injection in order to calculate renal clearance and to aid in the interpretation of whole-body clearance, including faecal excretion. Time-activity curves were generated for the thyroid, heart, brain, breasts and liver, by fitting the organ-specific geometric mean counts, obtained from regions of interest, to a multicompartmental model. The MIRD formulation, using 11 source organs, was applied to calculate the absorbed radiation doses for various organs upon administration of Z-[123I]MIVE. The images showed rapid hepatobiliary excretion which resulted in good imaging conditions for the thoracic region. Imaging of the abdominal region was impeded due to extensive bowel activity. Diffuse uptake and retention of activity was seen in breast tissue, the breast-to-non-specific uptake ratio increasing over time. Z-[123I]MIVE was cleared by both the kidneys and the gastrointestinal tract. At 50 h p.i. the mean excretion in urine was predicted to be 58%+/-14% (SD) and that in faeces 31%+/-19%. If the thyroid was not blocked, it was the most critical organ (0.33 mGy/MBq). In general, the excretory organs received the highest absorbed doses, i.e. the lower and upper large intestinal walls (0.11 and 0.098 mGy/MBq, respectively), the urinary bladder wall (0.090 mGy/MBq), the gallbladder wall (0.087 mGy/MBq) and the small intestine (0.043 mGy/MBq). The average effective dose equivalent of Z-[123I]MIVE was estimated to be 0.033 mSv/MBq. The amount of Z-[123I]MIVE required for adequate breast cancer ER imaging results in an acceptable effective dose equivalent to the patient. PMID- 9396874 TI - Relative renal uptake and transit time measurements using functional factor images and fuzzy regions of interest. AB - The aim of the study was a quantitative comparison of relative renal uptake and both the whole-kidney and the parenchymal transit time derived from factor analysis of image sequences and provided by standard clinical procedues. In order to extract the stable, well-interpretable factors, factor analysis was performed locally in the problem-specific time and spatial windows and the resulting factor images either evaluated directly as functional images or used as fuzzy regions of interest (ROIs) for the subsequent extraction of time-activity curves from the analysed data. The values of relative renal uptake of the left kidney measured in the functional factor images, which demonstrate the initial accumulation of activity in renal parenchyma (mean 51.0%), did not differ significantly from the values obtained by a standard method (mean 51.5%, r = 0.98, P<0.001). Whole kidney transit time calculated using fuzzy ROI curves correlated well with the reference values (r = 0.84, P<0.001); however, both its mean value (336.5 s) and the standard deviation (151.5 s) were substantially greater than those of the values provided by a standard procedure (262.8+/-86.9 s). Parenchymal transit time calculated using ROI curves correlated better with the transit time through a wider corticomedullary region rather than through a narrow cortical region, which is decisive in a differential diagnosis of renal disorders. In general, values of transit times provided by factor analysis correlated well with those provided by reference methods but with a shift towards the higher numerical values. This may have been a consequence of a greater extent of the automatically extracted fuzzy ROIs, or of occasionally delayed accumulation in the upper calyces. Results of the study provide quantitative evidence that the factor analysis of dynamic data, even without the introduction of prior physiological information, may yield clinically relevant information. However, some basic requirements, such as sufficiently high sampling frequency and count rate, adaption of the method to a specific clinical task, and proper selection of time and spatial windows for locally performed analysis, have to be fullfilled if the method is to be successfully applied clinically. PMID- 9396875 TI - Semi-automated renal region of interest selection method using the double threshold technique: inter-operator variability in quantitating 99mTc-MAG3 renal uptake. AB - In calculating the relative and absolute renal uptake of technetium-99m mercaptoacetyltriglycine (MAG3), inter-operator variability in the assignment of the renal region of interest (ROI) is a critical factor. Our goal was to develop a semi-automated method of assigning the renal ROI and then to compare the inter operator variability in calculating the percent injected dose (%ID) in the kidney at 1-2 min, using semi-automated versus manual ROIs. The manual ROIs were drawn independently by three operators (A, B and C). Operator A had about 20 years, experience in nuclear medicine, while operators B and C respectively had 3 years and 1 year of experience. In the semi-automated renal ROI selection method using the double-threshold technique, the operators only click around the centre of each kidney. The same three operators processed the ROIs using this double threshold method on 1-2 min images. The semi-automated method failed in three kidneys with very markedly reduced function owing to superimposition by liver or spleen. Inter-operator reproducibility in the remaining 59 kidneys was estimated using manual and semi-automated ROIs. With manual ROIs, the %ID (mean+/-standard error of mean) was 4.32+/-0.167 for A, 4. 14+/-0.165 for B and 3.28+/-0.139 for C. Although there was good correlation among them, these values were significantly different (P<0.0001). Using semi-automated ROIs, the %ID was 4.38+/ 0.160 for three operators. No significant difference was observed. Complete reproducibility was shown in 58 of 59 kidneys; the %ID difference of the remaining kidney was only 1.2%. The lowest %ID of all the kidneys successfully detected using the semi-automated method was 0. 77%. The semi-automated renal ROI selection method using the double-threshold technique displays good detectability of the renal contour. The renal uptake calculated using this method is reproducible and acceptable in routine clinical practice. PMID- 9396876 TI - Successful coronary revascularization improves prognosis in patients with previous myocardial infarction and evidence of viable myocardium at thallium-201 imaging. AB - The role of coronary revascularization of dysfunctional myocardium with preserved thallium-201 uptake in determining the prognosis in patients after myocardial infarction remains to be defined. This study was designed to evaluate the effects of successful revascularization on survival and left ventricular (LV) function in patients with previous myocardial infarction and evidence of dysfunctional but still viable myocardium at rest-redistribution 201Tl imaging. Seventy-six consecutive patients with LV dysfunction related to previous myocardial infarction and evidence of viable myocardium at rest-redistribution 201Tl tomography were followed for 17+/-8 months. LV ejection fraction (EF) was assessed by radionuclide angiography at baseline and after 13+/-2 months. Thirty nine patients were revascularized (group A) and 37 treated medically (group B). During the follow-up there were nine cardiac deaths. Survival rate was 97% in group A and 66% in group B (P<0.01). By Cox multivariate analysis, the extent of viable myocardium was the best predictor of cardiac death (chi2=8.67, P<0.01) and provided additional information to clinical and functional data (P<0.01). The inclusion of revascularization as a variable improved the global chi2 of the model from 14.1 to 21.9 (P<0.01). At follow-up, EF had improved by >/=5% in 16 patients. By multivariate logistic analysis, the extent of viable myocardium was the best predictor of EF improvement (chi2=15.49, P<0.001) and provided additional information to clinical and functional data (P<0.01). The inclusion of revascularization as a variable improved the global chi2 of the model from 16.8 to 22.5 (P<0.01). These results demonstrate that the total extent of dysfunctional myocardium with preserved 201Tl uptake is the strongest predictor of cardiac death in patients after myocardial infarction. Successful revascularization of dysfunctional but viable myocardium improves survival and LVEF in such patients. PMID- 9396877 TI - Comparison of dobutamine stress echocardiography and technetium-99m sestamibi single-photon emission tomography for the diagnosis of coronary artery disease in hypertensive patients with and without left ventricular hypertrophy. AB - Stress echocardiography has been considered an accurate method for the diagnosis of coronary artery disease in hypertensive patients and in patients with left ventricular hypertrophy. In contrast, the specificity of myocardial perfusion scintigraphy in these patients has been questioned. The aim of this study was to compare the accuracy of these two imaging modalities in conjunction with dobutamine stress test for the diagnosis of coronary artery disease in hypertensive patients with and without left ventricular hypertrophy. Dobutamine (up to 40 microg kg-1min-1) stress echocardiography in conjunction with sestamibi (MIBI) single-photon emission tomography (SPET) was performed in 84 patients with the diagnosis of systemic hypertension who had been referred for evaluation of myocardial ischaemia. Ischaemia was defined as new or worsened wall motion abnormalities at echocardiography and reversible perfusion defects at SPET. Significant coronary artery disease (>/=50% luminal diameter stenosis) was detected in 66 patients (79%). The sensitivity, specificity and accuracy of the ischaemic pattern at echocardiography for the diagnosis of coronary artery disease were 73% (CI 63%-82%), 83% (CI 75%-91%) and 75% (CI 66%-84%), those for MIBI were 67% (CI 57%-77%), 83% (CI 75%-91%) and 70% (CI 60%-80%) respectively (P = NS vs echocardiography). Significant stenosis was detected in 123 (49%) of the 252 analysed coronary arteries. The sensitivity, specificity and accuracy of echocardiography for the regional diagnosis of coronary artery disease were 63% (CI 56%-69%), 90% (CI 86%-94%) and 77% (CI 72%-82%). Those for MIBI were 58% (CI 51%-64%), 91% (CI 87%-94%) and 75% (CI 69%-80) respectively (P = NS vs echocardiography). Left ventricular hypertrophy was detected in 59 patients (70%) by echocardiography and did not influence the overall or regional specificity of echocardiography or MIBI SPET. It is concluded that in hypertensive patients, dobutamine stress echocardiography and MIBI SPET have a comparable accuracy for the overall and regional diagnosis of coronary artery disease. Hypertensive patients with or without left ventricular hypertrophy should not be considered unsuitable candidates for stress myocardial perfusion scintigraphy. PMID- 9396878 TI - Limited value of fluorine-18 fluorodeoxyglucose positron emission tomography for the imaging of neuroendocrine tumours. AB - Scintigraphy using [111In-DTPA-d-Phe1]-pentetreotide or pentavalent technetium 99m-dimercaptosuccinic acid [99mTc(V)-DMSA] has been shown to localize well differentiated and slowly growing neuroendocrine tumours, whereas increased fluorodeoxyglucose (FDG) uptake is associated with malignancy. The aim of this study was to compare the value of fluorine-18 FDG positron emission tomography (PET) with that of somatostatin receptor scintigraphy (SS-R) and dual radionuclide scintigraphy [SS-R and 99mTc(V)-DMSA = DNS] in detecting malignant neuroendocrine tumours. Fifteen patients with metastasizing gastroenteropancreatic tumours (GEP tumours; n = 7), medullary thyroid carcinomas (MTCs; n = 8) and elevated tumour markers [GEP tumours: 5-hydroxyindoleacetic acid, insulin; MTCs: calcitonin, carcinoembryonic antigen (CEA)] were studied. Prior to PET, all patients with GEP tumours underwent SS-R. DNS was performed in all patients with MTC. Patients had been fasting for at least 12 h and normal glucose plasma levels were confirmed. Sixty minutes after intravenous administration of 18F-FDG (mean: 374 MBq) whole-body PET and regional scans were performed. In addition, the resected tissues were prepared for immunocytochemistry examination (cell cycle-associated Ki-67 antigen). In two patients with less-differentiated GEP tumours associated with high proliferative activity and increased FDG uptake, SS-R failed to detect any lesion. In comparison, in four patients with well-differentiated GEP tumours showing low proliferative activity, SS-R localized four primary tumours, 22 lymph node metastases and 18 malignant liver lesions, whereas 18F-FDG PET demonstrated normal distribution. In one patient with a metastasizing carcinoid (medium proliferative activity) SS-R localized multiple metastases, whereas PET demonstrated low FDG uptake in all known metastases. In patients with recurrent MTC and rapidly increasing CEA levels DNS detected only three lesions in two patients, whereas PET demonstrated one pulmonary, three osseous, 20 mediastinal, ten locoregional, and four liver metastases in seven patients. Twenty-nine malignant lesions were confirmed by follow-up and nine lymph node metastases could be surgically removed. In conclusion, PET imaging of gastroenteropancreatic tumours revealed increased glucose metabolism only in less-differentiated GEP tumours with high proliferative activity and metastasizing MTC associated with rapidly increasing CEA levels. Therefore, additional 18F-FDG PET should be performed only if SS-R or DNS is negative. PMID- 9396879 TI - Design, construction and six years' experience of an integrated system for automated handling of discrete blood samples. AB - The present paper describes the design of an integrated system to aid in the taking and measurement of manual blood samples during nuclear medical examinations requiring blood sampling. In contrast to previously published systems, the present system is not used in the actual sampling of the blood, but aims to aid in all other aspects of handling and measurement. It consists of two main parts. One part is a distributed software system running on the scanner host computer used to register sample times, to display information pertaining to the ongoing examination and to collect data from a number of well crystals. The other main part consists of an industrial robot used to perform the actual weighing, centrifugation, pipetting and measurement of the samples. The system has been operational for 6 years, during which time it has had an "up-time" in excess of 95% and has handled and measured the blood samples from more than 5000 examinations, each comprising an average of 15 blood samples. The throughput of the system is 50 whole blood samples or 21 plasma samples per hour. In addition it has to a large extent removed the "human factor" from the process, thereby increasing the reliability of the data. PMID- 9396880 TI - Alpha-fetoprotein, free beta-human chorionic gonadotropin, and dimeric inhibin A produce the best results in a three-analyte, multiple-marker screening test for fetal Down syndrome. AB - OBJECTIVE: The purpose of this study was to determine, among six second-trimester maternal serum analytes, the best three-analyte combination for fetal Down syndrome detection. STUDY DESIGN: With use of commercially available assay kits, medians for free beta-human chorionic gonadotropin, CA 125, and dimeric inhibin A were established in stored sera from 45 to 50 euploid pregnancies at each week of gestation from 14 to 22 weeks and from 33 Down syndrome pregnancies. Maternal serum alpha-fetoprotein, unconjugated estriol, and intact human chorionic gonadotropin levels measured in each sample before storage were retrieved. All 20 possible three-analyte combinations were evaluated in the multiple-marker screening test for Down syndrome. RESULTS: The mean maternal age of the study population was 35.6 +/- 5.3 years. The best three-analyte combination was maternal serum alpha-fetoprotein, free beta-human chorionic gonadotropin, and dimeric inhibin A: 97% of Down syndrome cases were detected at a false-positive rate of 16%. At a slightly higher false-positive rate (18%) maternal serum alpha fetoprotein, estriol, and intact human chorionic gonadotropin detected only 79% of cases. CONCLUSIONS: Of six second-trimester maternal serum analytes, the best three-analyte combination for fetal Down syndrome detection was maternal serum alpha-fetoprotein, free beta-human chorionic gonadotropin, and dimeric inhibin A. This retrospective analysis should now be confirmed prospectively. PMID- 9396881 TI - Elevated second-trimester dimeric inhibin A levels identify Down syndrome pregnancies. AB - OBJECTIVE: Our purpose was to determine whether second-trimester dimeric inhibin A levels distinguish Down syndrome pregnancies from euploid pregnancies. STUDY DESIGN: With use of a commercially available enzyme-linked immunosorbent assay (Serotec, Oxford) inhibin A medians were established in stored sera from 40 to 50 euploid pregnancies at each week of gestation from 14 to 20 weeks and from 33 Down syndrome pregnancies. Maternal serum alpha-fetoprotein, unconjugated estriol, and human chorionic gonadotropin levels measured in each sample before storage were retrieved. The performance of inhibin A in the multiple-marker screening test was evaluated. RESULTS: The mean inhibin A multiple of the median was significantly higher in the Down syndrome group than in the euploid group (2.84 +/- 2.0 vs 1.22 +/- 1.0, p = 0.0001). An inhibin A level > or = 1.6 multiples of the median identified 70% of all Down syndrome pregnancies at a false-positive rate of 22%. Replacing estriol with inhibin A in the multiple marker screening test resulted in a higher Down syndrome detection rate at a lower screen-positive rate. CONCLUSION: Elevated second-trimester maternal serum inhibin A levels identify Down syndrome pregnancies; replacing estriol with inhibin A in the multiple-marker screening test improves test performance. PMID- 9396882 TI - Relationship between pregnancy-induced hypertension and placenta previa: a population-based study. AB - OBJECTIVE: Our purpose was to investigate, in a large population-based cohort, the hypothesis that the risk of pregnancy-induced hypertension is lower among pregnancies complicated by placenta previa compared with pregnancies occurring in women with fundally implanted placentas. STUDY DESIGN: Data for this retrospective cohort study were derived from the computerized Atlee perinatal database of the Reproductive Care Program, Nova Scotia, Canada. Women who were delivered in the province between 1980 and 1993 were included in the study. Patients with pregnancy-induced hypertension were clinically diagnosed by the presence of elevated blood pressure, proteinuria, or edema. The risk of pregnancy induced hypertension was compared between women diagnosed with placenta previa and those with a normally implanted placenta, after adjustment for potential confounders through multivariable logistic regression models based on the method of generalized estimating equations. RESULTS: During the 14 years (1980 to 1993), 121,082 singleton pregnancies were registered in the program, 416 (0.4%) of which had a confirmed diagnosis of placenta previa. Women with chronic hypertension had a relative risk of 1.2 (95% confidence interval 0.4 to 3.7) for placenta previa compared with normotensive women. However, the risk of pregnancy-induced hypertension was reduced by half among those with placenta previa (relative risk 0.5, 95% confidence interval 0.3 to 0.7). Adjustments for potential confounders, including maternal age, parity, prepregnancy body weight, prior cesarean delivery, prior spontaneous or induced abortions, and cigarette smoking, had no influence on this association. Analyses on the basis of stratification of women by parity (nulliparous vs multiparous), cigarette smoking (smoker vs nonsmoker), and gestational duration (< 28, 28 to 32, 33 to 36, and > 37 completed weeks) consistently showed reduced risks for pregnancy-induced hypertension among women with placenta previa, indicating that the association was not a result of shortened duration of gestation among women with placenta previa. CONCLUSIONS: The results from this study clearly show a decreased frequency of pregnancy induced hypertension among those pregnancies with placenta previa. We speculate that the pathophysiologic mechanisms for this finding may be due to altered placental perfusion seen among women diagnosed with placenta previa. PMID- 9396883 TI - Risk factors associated with preeclampsia in healthy nulliparous women. The Calcium for Preeclampsia Prevention (CPEP) Study Group. AB - OBJECTIVE: Our goal was to identify risk factors for the development of preeclampsia in nulliparous women enrolled in a multicenter trial comparing calcium supplementation to a placebo. STUDY DESIGN: A total of 4589 women from five centers was studied. Analysis of risk factors for preeclampsia was performed in 4314 who carried the pregnancy to > 20 weeks. Baseline systolic and diastolic blood pressure, demographic characteristics, and findings after randomization were examined for the prediction of preeclampsia. Preeclampsia was defined as hypertension (diastolic blood pressure > or = 90 mm Hg on two occasions 4 hours to 1 week apart) and proteinuria (> or = 300 mg/24 hours, a protein/creatinine ratio > or = 0.35, one dipstick measurement > or = 2+ or two dipstick measurements > or = 1+ at an interval as specified for diastolic blood pressure). RESULTS: Preeclampsia developed in 326 women (7.6%). The first analysis treated each risk factor as a categoric variable in a univariate regression. Maternal age, blood group and Rh factor, alcohol use, previous abortion or miscarriage, private insurance, and calcium supplementation were not statistically significant. Risk factors initially found to be significant were body mass index, systolic blood pressure, diastolic blood pressure, non-white race (African American and other), clinical center, and smoking. Adjusted odds ratios computed with a logistic regression model revealed that body mass index (odds ratio 3.22 for > or = 35 kg/m2 vs < 19.8 kg/m2), systolic blood pressure (odds ratio 2.66 for > or = 120 vs < 101 mm Hg), diastolic blood pressure (odds ratio 1.72 for > or = 61 mm Hg vs < 60 mm Hg), and clinical center (odds ratio 1.85 for Memphis vs the other clinical centers) were statistically significant predictors of preeclampsia. Results of the final model fit revealed that preeclampsia risk increases significantly (p < 0.0001) with increased body mass index at randomization, as well as with increased systolic and diastolic blood pressure at randomization. Calcium supplementation had no effect on the risks posed by body mass index and blood pressure. Among risk factors developing after randomization, an abnormal results of a glucose screen (plasma glucose > or = 140 mg/dl 1 hour after a 50 gm glucose challenge) was not found to be associated with a significant risk of preeclampsia. CONCLUSION: These risk factors should be of value in counseling women regarding preeclampsia and should aid in understanding the pathophysiologic characteristics of this syndrome. PMID- 9396884 TI - Better maternal outcomes are achieved with dexamethasone therapy for postpartum HELLP (hemolysis, elevated liver enzymes, and thrombocytopenia) syndrome. AB - OBJECTIVE: Our purpose was to determine whether the routine initiation of dexamethasone therapy in patients with postpartum HELLP (hemolysis, elevated liver enzymes, and thrombocytopenia) syndrome produces specific and general therapeutic benefits. STUDY DESIGN: In this retrospective, analytic study the puerperal courses of 43 women with postpartum HELLP syndrome who were treated with dexamethasone were compared with those of 237 similar patients who did not receive corticosteroids. Dexamethasone 10 mg intravenously at 12-hour intervals was given until disease remission was noted in treated patients, at which time up to two additional 5 mg intravenous doses were given at 12-hour intervals. RESULTS: The two patient groups were similar in regard to mode of delivery, gestational age, parity, and frequency of eclampsia. Compared with control subjects, dexamethasone-treated postpartum patients were more ill with significantly higher (p < 0.05) admission mean arterial blood pressure, higher serum uric acid level, and severe proteinuria. Dexamethasone administration was associated with a more rapid normalization of platelet counts and lactic dehydrogenase values. Most impressive was a clinically significant reduction of indicated transfusion and respiratory therapy, invasive hemodynamic monitoring, infectious or bleeding-related morbidity, and length of postpartum hospital course. CONCLUSIONS: Patients who received dexamethasone for postpartum-onset HELLP syndrome experienced a shorter disease course, faster recovery, less morbidity, and less need for other interventionist therapy compared with patients with HELLP syndrome who did not receive dexamethasone. PMID- 9396885 TI - A double-blind comparison of the safety and efficacy of intravaginal misoprostol and prostaglandin E2 to induce labor. AB - OBJECTIVE: Our purpose was to compare the safety and efficacy of intravaginally administered misoprostol versus prostaglandin E2 for labor induction in a double blind, randomized trial. STUDY DESIGN: One hundred three patients with indications for labor induction (including prelabor rupture of membranes) were randomized and received either misoprostol 50 micrograms or prostaglandin E2 (dinoprostone) 3 mg intravaginally. The dose was repeated 6, 24, and 30 hours after the first dose until active labor was achieved. For proper blinding, the drugs were prepared as identical-looking vaginal tablets. RESULTS: With use of a random number-generated table 52 patients were allocated to the misoprostol group and 51 to the prostaglandin E2 group. After exclusion of 3 patients, 50 in each group were evaluated. Delivery within 24 hours after administration occurred more often in the misoprostol group (70% vs 46% in the prostaglandin E2 group, p = 0.009), and fewer patients in this group needed more than two doses (12% vs 30%, p = 0.027). No difference in cesarean section rate (12% vs 14%, p = 0.67), fetal heart rate anomalies (33% vs 34%, p = 0.89), tachysystole (8% vs 14%, p = 0.37), hyperstimulation syndrome (0% vs 2%, not significant), meconium passage (28% vs 18%, p = 0.22), and fetal outcome (Apgar score at 1 and 5 minutes, arterial and venous umbilical cord blood pH, transfer to neonatal intensive care unit) was noted between the two groups. CONCLUSION: Intravaginal misoprostol is a safe drug for labor induction with superior effectiveness compared with intravaginal prostaglandin E2. PMID- 9396886 TI - International Multicentre Term Prelabor Rupture of Membranes Study: evaluation of predictors of clinical chorioamnionitis and postpartum fever in patients with prelabor rupture of membranes at term. AB - OBJECTIVES: Our purpose was to determine significant predictors for the development of clinical chorioamnionitis and postpartum fever in patients with prelabor rupture of membranes at term. STUDY DESIGN: Logistic regression analysis with odds ratios and 95% confidence intervals was used to determine the significant predictors of clinical chorioamnionitis and postpartum fever in women with prelabor rupture of membranes at term enrolled in this study. The study recently compared in a randomized controlled trial four strategies of management: induction with oxytocin, induction with prostaglandin, expectant management, and, if failed, induction with oxytocin or prostaglandin. RESULTS: The following variables were significantly associated with clinical chorioamnionitis: (1) number of digital vaginal examinations: > 8, 7 to 8, 5 to 6, 3 to 4 (vs 0 to 2) (odds ratio 5.07, 3.80, 2.62, 2.06); (2) duration of active labor: > or = 12, 9 to < 12, 6 to < 9 hours (vs < 3 hours) (odds ratio 4.12, 2.94, 1.97); (3) meconium-stained amniotic fluid (odds ratio 2.28); (4) parity of 0 (odds ratio 1.80); (5) time from membrane rupture to active labor: > or = 48, 24 to < 48 hours (vs < 12 hours) (odds ratio 1.76, 1.77); and (6) group B streptococcal colonization (odds ratio 1.71). Variables significantly associated with postpartum fever were (1) clinical chorioamnionitis (odds ratio 5.37), (2) duration of active labor: > or = 12, 9 to < 12, 6 to < 9, 2 to < 6 hours (vs < 3 hours) (odds ratio 4.86, 3.53, 3.46, 3.04), (3) cesarean section, operative vaginal delivery (odds ratio 3.97, 1.86), (4) group B streptococcal colonization (odds ratio 1.88), and (5) maternal antibiotics before delivery (odds ratio 1.94). CONCLUSIONS: Increasing numbers of digital vaginal examinations, longer duration of active labor, and meconium staining of the amniotic fluid were the most important risk factors for the development of clinical chorioamnionitis in women with prelabor rupture of membranes at term. The most important risk factors for the development of postpartum fever were clinical chorioamnionitis, increasing duration of active labor, and cesarean section delivery. PMID- 9396887 TI - Predictive factors for neonatal morbidity in neonates with an umbilical arterial cord pH less than 7.00. AB - OBJECTIVE: Fewer than 50% of neonates with an umbilical arterial pH < 7.00 have neonatal complications. Our objective was to identify clinical predictive factors for adverse outcomes in this group of neonates. STUDY DESIGN: In this case control study both cases and controls had an umbilical arterial cord pH < 7.00. Cases were defined as those neonates who had seizures, grade 3 to 4 intraventricular hemorrhage, gastrointestinal dysfunction, respiratory distress syndrome requiring intubation, sepsis, or death. Controls had an umbilical arterial cord pH < 7.00 and no complications. A multivariable prediction model was created, with variables having an association with adverse outcome by bivariate analyses, attempting to predict which neonates in this umbilical arterial pH range are at greatest risk for adverse outcomes. RESULTS: We identified 73 of 10,705 neonates born between July 1992 and October 1996 with an umbilical arterial cord pH < 7.00. Thirty-five neonates met our case definition, and the remaining 38 composed the control group. Cases had significantly lower arterial pH values and 1- and 5-minute Apgar scores, greater arterial base deficit values, and a higher incidence of abruptio placentae and maternal cocaine use. More cases were delivered before 34 weeks. There were three neonatal deaths, two cases of grade 3 or 4 intraventricular hemorrhage, five cases of gastrointestinal dysfunction, and four cases of neonatal seizures. In our predictive model for adverse neonatal outcome, an arterial base deficit > or = 16 mmol/L and a 5-minute Apgar score < 7 had a sensitivity and a specificity of 79% and 80.8%, respectively. CONCLUSION: Neonatal morbidity in neonates with an umbilical arterial cord pH < 7.00 can be predicted by a high arterial base deficit value and low 5-minute Apgar score. PMID- 9396888 TI - Managed care does not lower costs but may result in poorer outcomes for patients with gestational diabetes. AB - OBJECTIVE: Our purpose was to compare the costs of prenatal care and subsequent maternal and neonatal outcomes in patients with gestational diabetes cared for in an inner-city university hospital house staff clinic versus an inner-city managed care organization. STUDY DESIGN: A retrospective cohort study was conducted. The groups consisted of 115 patients with gestational diabetes who were cared for in a house staff clinic and a demographically similar group of 85 patients cared for in a neighborhood managed care organization. The groups were examined regarding baseline demographics, intensity of prenatal care, maternal and neonatal outcomes, and total cost of the provision of care. RESULTS: There was no difference between groups in the total cost of maternal-infant care. A larger percentage of patients in the house staff group saw the physician frequently. In contrast, patients cared for in the managed care organization underwent more tests of fetal well-being. There was a greater rate of neonatal macrosomia in the managed care organization group compared with the house staff group. CONCLUSIONS: Managed care does not decrease the cost of caring for patients with gestational diabetes but does lead to a greater rate of neonatal macrosomia, which may reflect poorer glucose control. PMID- 9396889 TI - Fractured clavicle and Erb's palsy unrelated to birth trauma. AB - OBJECTIVES: Our purpose was to determine the perinatal factors associated with clavicular fracture or Erb's palsy in neonates and to document the percentage of cases where no risk factors were involved. STUDY DESIGN: We reviewed the medical records of all live-born singleton infants admitted to the newborn nurseries between 1992 and 1995. Mothers and infants with clavicular fracture or Erb's palsy were compared with those without these birth injuries. RESULTS: Of 11,636 neonates, there were 236 (2.03%) with clavicular fracture and 51 (0.44%) with Erb's palsy. Clavicular fracture was significantly associated with shoulder dystocia and high birth weight. Significant factors associated with Erb's palsy were shoulder dystocia, high birth weight, prolonged second stage, instrumental delivery, fetal distress, use of oxytocin, and epidural anesthesia. A total of 51.7% of the neonates with clavicular fracture and 29.4% of those with Erb's palsy had none of the risk factors examined. CONCLUSIONS: Although macrosomic fetuses and instrumental or difficult deliveries are risk factors for clavicular fracture and Erb's palsy, > 50% and 25%, respectively, occur without the risk factors examined. PMID- 9396890 TI - The accuracy of the summated amniotic fluid index in evaluating amniotic fluid volume in twin pregnancies. AB - OBJECTIVE: Our purpose was to determine the accuracy of the summated amniotic fluid index designed to estimate the total amniotic fluid volume in twin pregnancies. STUDY DESIGN: The summated amniotic fluid index was measured in 62 normal diamniotic twin pregnancies by adding the deepest vertical pockets in the four quadrants. Actual amniotic fluid volume was then determined in all 124 amniotic sacs by amniocentesis and a dye-dilution technique. For data analysis, amniotic fluid volumes were classified by percentile with use of previously reported norms. RESULTS: There were significant differences in the percentile distribution of amniotic fluid volume as estimated by the summated amniotic fluid index and the actual volume as determined by dye dilution (p < 0.001). The summated amniotic fluid index has a sensitivity of only 13% in predicting amniotic sac volume. CONCLUSION: The summated amniotic fluid index is a poor predictor of intertwin differences in amniotic fluid volume and cannot identify twin pairs at risk for oligohydramnios and hydramnios. PMID- 9396891 TI - Monoamniotic twins: improved perinatal survival with accurate prenatal diagnosis and antenatal fetal surveillance. AB - OBJECTIVE: Our goal was to report our 10-year experience with monoamniotic twins and to compare that experience with cases reported in the literature. STUDY DESIGN: Records of all monoamniotic twin pregnancies managed at the University of Connecticut Health Center from March 1986 to August 1996 were reviewed. A MEDLINE search from January 1966 to August 1996 was performed, and each report was screened for accuracy of diagnosis. Only cases with umbilical cord entanglement of nonconjoined like-sex twins, the obstetrician's confirmation at delivery, or pathologic confirmation of monoamniotic placentation were included. Data collected were as follows: birth outcome, gestational age at delivery, birth weight, gender, Apgar scores, hematocrit, cord knotting, and neonatal complications. Cases from the literature were divided into those with prenatal diagnosis and those without. RESULTS: Thirteen monoamniotic pregnancies resulting in 26 infants who were born alive were managed at our center. The average gestational age at diagnosis was 16.3 weeks. All had antenatal fetal surveillance including serial sonograms and nonstress tests. The average gestational age and birth weight at delivery were 32.9 weeks and 1669 gm, respectively. Cord entanglement was noted in all cases, with knotting in 8 of 13. Two pairs of 26 newborns had evidence of twin-twin transfusion syndrome. Eight of 13 monoamniotic pregnancies were delivered because of nonreassuring results of nonstress test, two because of preterm labor, two electively because of lung maturity, and one because of intrauterine growth restriction. Two of the 26 infants died in the neonatal period, one of congenital heart disease and one of sepsis and asphyxia. The MEDLINE search revealed 96 articles with a total of 202 sets of monoamniotic twins. Comparison of cases (13 sets) with the historic control group without prenatal diagnosis (77 sets) showed a 71% reduction in relative risk of perinatal mortality. CONCLUSIONS: With accurate prenatal diagnosis, intensive fetal surveillance, and appropriately timed delivery, perinatal survival of monoamniotic twins is improved; it was 92% in this series. PMID- 9396892 TI - Is size discordancy an indication for delivery of preterm twins? AB - OBJECTIVE: Our goal was to determine the clinical significance of size discordancy in preterm twins. STUDY DESIGN: A retrospective study was performed to review outcomes of twins delivered between Jan. 1, 1988, and June 30, 1995. Maternal and neonatal records were assessed for demographic data, maternal medical history, and neonatal mortality and morbidity outcomes. Discordancy was defined as > or = 20% difference in birth weight. The chi 2 analysis was performed. RESULTS: There were 42 sets of discordant twins and 77 sets of concordant twins in the final analysis. The distribution of gestational ages in both groups was similar. We found no difference in maternal morbidity between the groups. Discordant sets had a significantly longer hospital stay (p = 0.003) and more cases of hyperbilirubinemia (p = 0.01), but there were no other differences in morbid outcomes. There were no differences in outcome variables between the two twins within discordant sets with respect to gender, size, birth order, growth restriction, or route of delivery. There were no stillbirths among any of the 238 infants. Of the 15 neonatal deaths, none occurred in infants delivered after 32 weeks' gestation or in infants weighing > 2000 gm at birth. Infants who were small for gestational age had a higher incidence of sepsis (p = 0.043) and longer hospital stays (p = 0.005) compared with infants who were appropriate for gestational age. CONCLUSIONS: Size discordancy alone does not appear to be an indication for preterm delivery of twins. When results of antenatal testing are normal and growth restriction is absent, attempts should be made to achieve a gestational age > 32 weeks and weight > 2000 gm before delivery is considered. PMID- 9396893 TI - Critical periods of maternal weight gain: effect on twin birth weight. AB - OBJECTIVE: Our purpose was to evaluate the association between maternal weight gain and twin birth weight. STUDY DESIGN: This historic cohort study was based on 646 live-born twin births of > or = 28 weeks from Baltimore, Maryland, Miami, Florida, and Ann Arbor, Michigan. The sum of twin-pair birth weight was modeled as a function of either net maternal weight at delivery or rates of maternal weight gain with use of multiple regression. RESULTS: Birth weight was significantly associated with weight gain before 20 weeks in underweight women, before 20 weeks and after 28 weeks in overweight women, and during all three gestational periods in normal-weight women. Weight gain before 20 weeks had the largest effect on infants of underweight women, less of an effect on infants of normal-weight women, and half as much effect on infants of overweight women. Weight gain after 28 weeks significantly affected the infant birth weights of normal-weight and overweight women, but the effect was half as great among infants of the latter group. CONCLUSIONS: These findings suggest that weight gain during critical periods of gestation significantly influences twin birth weight; these critical periods vary by maternal pregravid weight status. PMID- 9396894 TI - The relationship of infection to method of delivery in twin pregnancy. AB - OBJECTIVE: Our purpose was to determine whether manipulation of the second twin increases the risk of postpartum infection. STUDY DESIGN: Medical records of all twin deliveries between January 1991 and December 1994 were reviewed. The route of delivery (vaginal vs cesarean section) was examined. The vaginal group was further divided into those delivered in the vertex/vertex position (i.e., no uterine manipulation) versus those delivered vertex/breech extraction (i.e., manipulation). The chi 2 and Student t test were used where appropriate. RESULTS: A total of 718 twins were identified, and maternal age, parity, gestational age at delivery (36 weeks), and birth weight (2278 gm) were similar among groups. The metritis rate was higher in the cesarean group (74/447 or 18%) than in the vaginal group (17/299 or 5.7%, p < 0.001). In comparing the vaginal group delivered without uterine manipulation with the vaginal group delivered with manipulation (i.e., breech extraction), there was no difference in the incidence of metritis (10/147 or 6.8% vs 7/152 or 4.6%, not significant). The length of time between delivery of twin A and twin B did not affect the metritis rate. Neonatal outcomes including sepsis, neonatal death, and length of hospitalization were similar among groups (not significant). CONCLUSION: Uterine manipulation of the second twin does not increase the risk of postpartum metritis or neonatal sepsis. In addition, the time interval between delivery of twins A and B has no effect on the rate of metritis. Although the rate of endometritis has been reported to be higher with twins delivered by cesarean section compared to singletons, the 18% rate of endometritis in twins delivered by cesarean section in this study is slightly lower than in our general population of cesarean deliveries (22%). PMID- 9396895 TI - A randomized clinical trial comparing misoprostol with prostaglandin E2 gel for preinduction cervical ripening. AB - OBJECTIVE: Our purpose was to perform a randomized trial comparing intravaginal misoprostol to intravaginal prostaglandin E2 gel for preinduction cervical ripening evaluating efficacy and side effects. STUDY DESIGN: Seventy-five women seen for induction of labor were randomized to receive 100 micrograms of intravaginal misoprostol or 5 mg of pharmacy-prepared intravaginal prostaglandin E2 gel for cervical ripening before oxytocin induction. Six hours after placement of the study agent, patients were given oxytocin if they were not in labor. The primary outcome measure was induction-to-delivery time; secondary measures were change in Bishop score, delivery mode, and side effects. Results were analyzed by the Student t test and Fisher's exact test, with p < 0.05 considered significant. RESULTS: There was no difference in the incidence of primiparity or the median initial Bishop score between the two study groups. The mean time to delivery and the need for oxytocin was significantly less for subjects receiving misoprostol. There was no difference in the incidence of uterine hyperstimulation syndrome or cesarean delivery between the groups. CONCLUSIONS: This randomized clinical trial indicates that misoprostol is efficacious for preinduction cervical ripening. Misoprostol use resulted in a significantly shorter induction-to-delivery time compared with prostaglandin E2 gel use. The side effects associated with misoprostol may be dose related, and further studies to identify the optimum dosage and interval are needed. PMID- 9396896 TI - The association of placenta previa with history of cesarean delivery and abortion: a metaanalysis. AB - OBJECTIVE: Our purpose was to determine the incidence of placenta previa based on the available epidemiologic evidence and to quantify the risk of placenta previa based on the presence and number of cesarean deliveries and a history of spontaneous and induced abortion. STUDY DESIGN: We reviewed studies on placenta previa published between 1950 and 1996 on the basis of a comprehensive literature search with use of MEDLINE and by identifying studies cited in the references of published reports. Studies were chosen for inclusion in the metaanalysis if the incidence of placenta previa and its cross-classification with either prior cesarean delivery or abortions (both spontaneous and induced) or both were available. We also extracted details about the study design (case-control or cohort study) and place where they were conducted (United States or other countries). Published case reports dealing with placenta previa and studies relating to abruptio placentae were excluded from this review. We also restricted the search to studies published in English. No attempts were made to locate any unpublished studies. Data from studies identified during the literature search were reviewed and abstracted by a single author. In case of discrepancies or when the information presented in a study was unclear, abstraction by a (blinded) second reviewer was sought to resolve the discrepancy. RESULTS: Data on the incidence of placenta previa and its associations with previous cesarean delivery and abortions were abstracted. Subgroup analyses were performed to identify potential sources of heterogeneity by study design and place where they were conducted. Statistical methods used for the metaanalysis included the fixed effects logistic regression model, whereas potential sources of heterogeneity among studies were evaluated by fitting random-effects models. The tabulation of 36 studies identified a total of 3.7 million pregnant women, of whom 13,992 patients were diagnosed with placenta previa. The reported incidence of placenta previa ranged between 0.28% and 2.0%, or approximately 1 in 200 deliveries. Women with at least one prior cesarean delivery were 2.6 (95% confidence interval 2.3 to 3.0) times at greater risk for development of placenta previa in a subsequent pregnancy. The results varied by study design, with case-control studies showing a stronger relative risk (relative risk 3.8, 95% confidence interval 2.3 to 6.4) than cohort studies did (relative risk 2.4, 95% confidence interval 2.1 to 2.8). Four studies, encompassing 170,640 pregnant women, provided data on the number of previous cesarean deliveries. These studies showed a dose-response pattern for the risk of previa on the basis of the number of prior cesarean deliveries. Relative risks were 4.5 (95% confidence interval 3.6 to 5.5) for one, 7.4 (95% confidence interval 7.1 to 7.7) for two, 6.5 (95% confidence interval 3.6 to 11.6) for three, and 44.9 (95% confidence interval 13.5 to 149.5) for four or more prior cesarean deliveries. Women with a history of spontaneous or induced abortion had a relative risk of placenta previa of 1.6 (95% confidence interval 1.0 to 2.6) and 1.7 (95% confidence interval 1.0 to 2.9), respectively. Substantial heterogeneity in the results of the metaanalysis was noted among studies. CONCLUSION: There is a strong association between having a previous cesarean delivery, spontaneous or induced abortion, and the subsequent development of placenta previa. The risk increases with number of prior cesarean deliveries. Pregnant women with a history of cesarean delivery or abortion must be regarded as high risk for placenta previa and must be monitored carefully. This study provides yet another reason for reducing the rate of primary cesarean delivery and for advocating vaginal birth for women with prior cesarean delivery. PMID- 9396897 TI - Neonatal nucleated red blood cell counts in small-for-gestational age fetuses with abnormal umbilical artery Doppler studies. AB - OBJECTIVE: The presence of elevated nucleated red blood cell counts in neonatal blood has been associated with fetal hypoxia. We sought to determine whether small-for-gestational-age fetuses with abnormal umbilical artery Doppler velocity waveforms have elevated nucleated red blood cell counts. STUDY DESIGN: Hospital charts of neonates with the discharge diagnosis of small for gestational age (birth weight < 10th percentile) who were delivered between October 1988 and June 1995 were reviewed for antepartum testing, delivery conditions, and neonatal outcome. We studied fetuses who had an umbilical artery systolic/diastolic ratio within 3 days of delivery and a complete blood cell count on the first day of life. Multiple gestations, anomalous fetuses, and infants of diabetic mothers were excluded. Statistical analysis included the Student t test, chi 2 analysis, analysis of variance, and simple and stepwise regression. RESULTS: Fifty-two infants met the inclusion criteria. Those with absent or reversed end-diastolic velocity (n = 19) had significantly greater nucleated red blood cell counts than did those with end-diastolic velocity present (n = 33) (nucleated red blood cells/100 nucleated cells +/- SD: 135.5 +/- 138 vs 17.4 +/- 23.7, p < 0.0001). These infants exhibited significantly longer time intervals for clearance of nucleated red blood cells from their circulation (p < 0.0001). They also had lower birth weights (p < 0.05), lower initial platelet count (p = 0.0006), lower arterial cord blood pH (p < 0.05), higher cord blood base deficit (p < 0.05), and an increased likelihood of cesarean section for "fetal distress" (p < 0.05). Multivariate analysis demonstrated that absent or reversed end-diastolic velocity (p < 0.0001) and low birth weight (p < 0.0001) contributed to the elevation of the nucleated red blood cell count, whereas gestational age at delivery was not a significant contributor. CONCLUSION: We observed significantly greater nucleated red blood cell counts and lower platelet counts in small-for-gestational-age fetuses with abnormal umbilical artery Doppler studies. This may suggest that antenatal thrombotic events lead to an increased placental impedance. Fetal response to this chronic condition may result in an increased nucleated red blood cell count. PMID- 9396898 TI - Elevated maternal serum midtrimester alpha-fetoprotein levels are associated with fetoplacental ischemia. AB - OBJECTIVE: Elevation of maternal serum alpha-fetoprotein in the second trimester is associated with poor pregnancy outcome, including fetal death, preterm delivery, and fetal growth restriction. We hypothesized that placental ischemia may be the common underlying pathogenesis of these outcomes. Thus we tested angiogenin, a potent inducer of neovascularization, in midtrimester amniotic fluid of patients with elevated maternal serum alpha-fetoprotein values to determine whether alpha-fetoprotein elevation is due to ischemia with subsequent stimulation of angiogenesis. STUDY DESIGN: In this case-control study, patients with elevated maternal serum alpha-fetoprotein levels (> or = 2.0 multiples of the median, n = 9) at triple screen were matched with two controls (n = 18) on the basis of year of amniocentesis and maternal age, race, and parity. The median elevation of maternal serum alpha-fetoprotein in the study population was 4.01 multiples of the median (range 2.65 to 7.24 multiples of the median). Inclusion criteria were (1) singleton gestation, (2) no evidence of fetal structural or chromosomal anomalies, and (3) genetic amniocentesis. Amniotic fluid was immunoassayed for angiogenin (Quantikine, R&D Systems; sensitivity 0.026 ng/ml, interassay and intraassay coefficients of variation 4.6% and 2.9%, respectively). Statistical analysis included one-way analysis of variance and regression with p < 0.05 significant. Angiogenin and maternal serum alpha-fetoprotein values were normalized with use of natural log transformation for statistical analysis. RESULTS: Angiogenin values were significantly elevated in patients with high maternal serum alpha-fetoprotein levels (median 31.1 [range 9.2 to 54.6] vs 17.1 [range 9.0 to 29.2] ng/ml, p = 0.02). Mean gestational age at sampling, maternal age, and year of amniocentesis were not significantly different between the study and control groups (each p > 0.05). As anticipated, there was a significant increase in preterm deliveries and small-for-gestational-age neonates in the patients with elevated maternal serum alpha-fetoprotein levels (each p < 0.01). CONCLUSIONS: Midtrimester amniotic fluid angiogenin levels are significantly elevated in patients with elevated midtrimester maternal serum alpha-fetoprotein levels. Because angiogenin is a known marker of tissue ischemia, resulting in neovascularization, we hypothesize that elevation of maternal serum alpha fetoprotein levels at triple screen is due to placental ischemia. PMID- 9396899 TI - Fetoplacental vascular tone during fetal circuit acidosis and acidosis with hypoxia in the ex vivo perfused human placental cotyledon. AB - OBJECTIVES: Our purpose was to determine the effects of acidosis and acidosis hypoxia on fetoplacental perfusion pressure and its response to angiotensin II. STUDY DESIGN: Perfused cotyledons from 14 placentas were studied with either an acidotic fetal circuit perfusate (n = 7) or an acidotic-hypoxic fetal circuit perfusate (n = 7). Each cotyledon's fetal vasculature was initially perfused under standard conditions and bolus injected with 1 x 10(-10) moles of angiotensin II. Fetoplacental perfusate was then replaced with either an acidotic medium (pH 6.90 to 7.00 and Po2 516 to 613 mm Hg) or an acidotic-hypoxic medium (pH 6.90 to 7.00 and Po2 20 to 25 mm Hg) followed by an angiotensin II injection. The vasculature was subsequently recovered with standard perfusate and again injected with angiotensin II. Perfusion pressures within each group were compared by one-way analysis of variance, and results were expressed as mean pressure +/- SEM. RESULTS: Resting fetoplacental perfusion pressure did not change when the fetal circuit perfusate was made acidotic (28 +/- 1 mm Hg vs 25 +/- 2 mm Hg) or acidotic-hypoxic (26 +/- 2 mm Hg vs 25 +/- 2 mm Hg). The maximal fetoplacental perfusion pressure achieved in response to angiotensin II did not differ with an acidotic perfusate (41 +/- 2 mm Hg vs 38 +/- 1 mm Hg) or with an acidotic-hypoxic perfusate (39 +/- 2 mm Hg vs 36 +/- 2 mm Hg). CONCLUSIONS: In the perfused placental cotyledon fetoplacental perfusion pressure and pressor response to angiotensin II are not affected by fetal circuit acidosis or acidosis-hypoxia. This suggests that neither fetal acidosis nor fetal acidosis combined with hypoxia has a direct effect on fetoplacental vascular tone. PMID- 9396900 TI - The fetoplacental pressor effects of low-dose acetylsalicylic acid and angiotensin II in the ex vivo cotyledon model. AB - OBJECTIVE: Our purpose was to investigate perfusion pressure changes ex vivo induced by angiotensin II on fetoplacental vasculature pretreated with low-dose acetylsalicylic acid. STUDY DESIGN: Two cotyledons from each of 12 placentas were perfused. The intervillous space of one cotyledon was infused with acetylsalicylic acid (5 x 10(-5) mol/L) similar to the serum concentration of women receiving daily low-dose aspirin therapy (60 to 81 mg). The control cotyledon was infused with an equivalent amount of normal saline solution. Two doses of angiotensin II, 1 x 10(-11.5) and 1 x 10(-10) moles, were injected as boluses into the chorionic arteries of each cotyledon. A 3 x 10(-7) mole dose of angiotensin II was also injected into the intervillous space. Statistical analysis was performed with analysis of variance, and results are expressed as mean pressure change in millimeters of mercury +/- SEM. RESULTS: Perfusion pressure response did not vary between cotyledons pretreated with acetylsalicylic acid and control cotyledons when 3 x 10(-7) moles of angiotensin II was injected into the intervillous space (8.0 +/- 1.9 mm Hg vs 9.8 +/- 1.6 mm Hg, p = 0.59). There were no differences between cotyledons in pressure response to 1 x 10( 11.5) moles of angiotensin II injected into the fetal circuit (5.9 +/- 0.8 mm Hg vs 6.7 +/- 0.9 mm Hg, p = 0.51). However, in the cotyledons pretreated with acetylsalicylic acid there was a decrease in the pressor response to 1 x 10(-10) moles of angiotensin II (14.1 +/- 1.4 mm Hg vs 21.5 +/- 3.3 mm Hg, p = 0.05). CONCLUSIONS: Low-dose aspirin infused into the intervillous space decreases vasoconstriction elicited by angiotensin II in the fetoplacental compartment. This suggests that maternal low-dose aspirin therapy has effects in the fetoplacental circulation in addition to its effects in the maternal circulation. PMID- 9396901 TI - The association between maternal cocaine use and placenta previa. AB - OBJECTIVE: Our aim was to determine whether maternal cocaine exposure is a risk factor for placenta previa. STUDY DESIGN: In this case-control study, cases of placenta previa confirmed at delivery (ascertained by International Classification of Diseases, ninth revision, Clinical Modification, code-based search, N = 40) were compared with a random sample of patients without placenta previa (N = 80) in a ratio of two controls per case. Data on antecedent maternal cocaine use, as well as other potential risk factors for placenta previa, were obtained from a review of the prenatal chart and the hospital record. Categorization of cocaine use was based on either patient self-report or urine toxicologic testing, or both. Multiple logistic regression was performed to assess the association between cocaine and placenta previa while we controlled for other variables. RESULTS: After the effects of other variables were adjusted for, maternal cocaine use was an independent risk factor for placenta previa (adjusted odds ratio = 4.39, 95% confidence interval 1.17 to 16.4). Other significant risk factors included a history of cesarean section and prior elective abortion. CONCLUSION: These results suggest that cocaine use, as well as prior cesarean section, prior elective abortion, and parity, are associated with placenta previa. PMID- 9396902 TI - New birth weight nomograms for twin gestation on the basis of accurate gestational age. AB - OBJECTIVE: Our purpose was to establish new nomograms for the birth weight of twins on the basis of accurate methods to validate gestational age. STUDY DESIGN: The medical records of 1632 consecutive twin gestations delivered between 1984 and 1996 were reviewed. Only pregnancies induced by ovulation induction techniques or that were measured ultrasonographically for crown-rump length during the first trimester were included. Excluded were those whose fetuses (one or both) were stillborn, or if the mother smoked, had a significant chronic illness, or was prescribed any regular medications. The study comprised 520 twin pregnancies at 28 to 41 gestational weeks at delivery. RESULTS: The median and 10th and 90th percentile birth weight curves were calculated for the studied twins and plotted against previously reported singleton nomograms. Fetuses of twin pregnancies were found to be growth restricted in comparison with previously reported singletons throughout the third trimester. This trend became more evident after the thirty-fourth to thirty-sixth weeks. CONCLUSIONS: We recommend these novel birth weight nomograms for clinical use in the management of twin pregnancies. PMID- 9396903 TI - Anticholinergic suppression of ovine fetal swallowing activity. AB - OBJECTIVE: Fetal swallowing contributes importantly to amniotic fluid volume regulation as the primary route of fluid resorption, reaching 500 to 1000 ml/day near term. Near-term ovine fetal swallowing activity occurs predominantly during low-voltage electrocortical activity. In view of the potential to pharmacologically alter electrocortical activity, we hypothesized that fetal administration of a centrally acting cholinergic antagonist may be used to modulate fetal swallowing activity. To explore cholinergic modulation of swallowing activity, we examined fetal swallowing and electrocortical activity in response to central and peripheral cholinergic suppression by atropine sulfate. STUDY DESIGN: Singleton ovine fetuses (n = 6) were chronically prepared with vascular catheters and thyrohyoid, nuchal, and thoracic esophageal electromyogram and biparietal electrocortical electrodes. Swallowing and electrocortical activity were monitored for 2 hours before and after intravenous injection (1 ml of 0.15 mol/L sodium chloride) of atropine sulfate (1 mg/kg). On a subsequent day an identical study was performed with use off atropine methyl nitrate (3 mg/kg), an atropine analog that does not cross the blood-brain barrier. RESULTS: Atropine sulfate decreased low-voltage electrocortical activity (56% +/- 5% to 14% +/- 4%), increased high-voltage electrocortical activity (40% +/- 5% to 81% +/- 5%), and did not change intermediate electrocortical activity (4% +/- 1% to 5% +/- 1%). Fetal swallowing activity decreased from 46 +/- 12 to 12 +/- 2 swallows per hour after atropine sulfate administration. Atropine methyl nitrate had no discernible effect on either fetal electrocortical or swallowing activity. Fetal arterial pressure, plasma osmolality, pH, PCO2, and PO2 did not change. CONCLUSIONS: Central cholinergic antagonism suppresses low-voltage fetal electrocortical and swallowing activity in the ovine fetus. Studies exploring spontaneous or induced fetal swallowing should consider the behavioral state of the fetus when conclusions are drawn about changes in the swallowing activity. PMID- 9396904 TI - Should the same glucose values be targeted for type 1 as for type 2 diabetics in pregnancy? AB - OBJECTIVE: Our purpose was to determine whether the same maternal glycemic control is necessary to achieve similar perinatal outcomes for type 1 as for type 2 diabetics. STUDY DESIGN: The subjects were all women with pregestational diabetes mellitus delivered of live-born singletons. Glycemic control was achieved with diet and insulin. Self-monitoring of blood glucose was performed before meals and at bedtime. Target glucose values were 60 to 90 mg/dl fasting and 60 to 105 mg/dl at other times. RESULTS: Of 60,628 deliveries, 46 type 1 and 113 type 2 diabetic women met inclusion criteria. Respective differences were found between type 1 and type 2 diabetics in average daily glucose levels (112 mg/dl vs 97 mg/dl, p < 0.001), percent of values within target ranges (35% vs 57%, p < 0.001), and mean amplitude of glycemic excursion (48.1 mg/dl vs 24.9 mg/dl, p < 0.001). At least one daily glucose value was < 50 mg/dl during 19% of observation days for type 1 vs 2% of observation days for type 2 pregnancies (p < 0.001). There were no statistically significant differences between type 1 and type 2 diabetic pregnancies in neonatal macrosomia (30% vs 34%), proportion of cesarean deliveries during labor for arrest disorders (67% vs 69%), shoulder dystocia (2% vs 6%), and neonatal hypoglycemia (18% vs 26%). CONCLUSIONS: Less stringent maternal glycemic control may permit comparable maternal and neonatal outcomes for type 1 compared with type 2 diabetics. Higher target values for type 1 diabetics may decrease the frequency of maternal hypoglycemic episodes. PMID- 9396905 TI - Glucometer analysis of one-hour glucose challenge samples. AB - OBJECTIVE: Our purpose was to explore a cost-saving measure for diabetes screening by using glucometer testing on venous whole blood obtained after a 1 hour glucose challenge test. Glucometer results falling above an upper threshold would predict abnormal plasma values and mandate a glucose tolerance test; results below the lower threshold would predict normal plasma values and avoid further testing. Results between the thresholds would require traditional plasma analysis. STUDY DESIGN: We performed a prospective cohort study on 222 consecutive pregnant women. A standard 50 gm glucose screen was performed with venous blood drawn at 1 hour. We immediately removed a drop of whole blood from the venous sample and analyzed it on a portable glucometer, Accu-Chek III. The remaining sample was submitted immediately for routine plasma analysis. All values were obtained on the same glucometer, which was calibrated daily in our clinic laboratory. Regression analysis was performed on 129 samples to select the two thresholds. The selected thresholds were then applied prospectively to the next 93 consecutive samples for validation. RESULTS: Excellent correlation (r = 0.9045) exists between the glucometer and laboratory values. Glucometer threshold values of 110 mg/dl and 155 mg/dl were selected because they predicted plasma values < 135 mg/dl or > 135 mg/dl with 95% certainty, respectively. Prospectively, the thresholds were completely accurate in classifying the values. CONCLUSION: Venous whole blood assayed by glucometer can reliably predict an elevated or normal automated plasma glucose value. By applying thresholds, three fourths of all patients can immediately receive reassuring information, whereas the patients with poorest glucose tolerance are immediately identified and diagnostic testing is scheduled. Additionally, our model reduces the number of automated laboratory studies by 80% and reduces the cost of diabetic screening. PMID- 9396906 TI - The effect of smoking tobacco on neonatal body composition. AB - OBJECTIVE: Our purpose was to examine the differences in body composition in infants of women who smoke compared with those of nonsmokers. STUDY DESIGN: Within 24 hours of birth anthropometric measurements and total body electrical conductivity estimates of body composition were obtained on 129 term infants (30 born to women who smoked tobacco during pregnancy and 99 born to women who did not smoke). Anthropometric measurements included weight, skinfolds, circumferences, and crown-heel and extremity lengths. RESULTS: There was a significant decrease in weight in the infants of smokers (3145 +/- 414 gm vs 3354 +/- 525 gm, p = 0.05). There was a decrease in crown-heel length (49.2 +/- 2.0 cm vs 50.1 +/- 2.2 cm, p = 0.03), the length of the lower leg (7.9 +/- 0.6 cm vs 8.4 +/- 0.6 cm, p = 0.0001), upper leg (9.1 +/- 0.7 cm vs 9.9 +/- 0.8 cm, p = 0.0001), and the lower arm (7.2 +/- 0.5 cm vs 7.5 +/- 0.4 cm, p = 0.0001). There were no significant differences in the skinfold and limb circumference measurements. Infants of smokers had significantly decreased fat-free mass as estimated by total body electrical conductivity (2799 +/- 292 gm vs 2965 +/- 359 gm, p = 0.02) but no significant difference in fat mass (343 +/- 164 gm vs 387 +/ 216 gm, p = 0.32). CONCLUSIONS: The decrease in birth weight in infants of women who smoked is primarily due to a decrease in fat-free mass. PMID- 9396907 TI - Peripartum cardiomyopathy: a longitudinal echocardiographic study. AB - OBJECTIVE: Our purpose was to determine echocardiographic trends after initial diagnosis of peripartum cardiomyopathy. STUDY DESIGN: Nine women diagnosed with peripartum cardiomyopathy were prospectively recruited for a longitudinal echocardiographic study. Severe myocardial dysfunction was defined as left ventricular end-diastolic dimension > or = 60 mm + fractional shortening < or = 21%, and mild dysfunction was defined as left ventricular end-diastolic dimension < 60 mm + fractional shortening 22% to 24%. Unpaired t tests were used to compare sample means and Fisher's exact test used to compare discrete variables. RESULTS: All women were seen initially for pulmonary edema. Echocardiography showed decreased systolic function in all women. The mean age at diagnosis was 33.0 +/- 6.9 years. All but one woman had a diagnosis of either chronic hypertension (n = 6) or preeclampsia (n = 2). Four women were first seen ante partum and five post partum (range 1 day to 2 months). Repeat echocardiography was performed in all nine women (median 8 months, range 6 weeks to 5 years). There was no correlation between antepartum or postpartum presentation and cardiovascular status on follow up (p = 0.3). Values for initial left ventricular end-diastolic dimension, severe versus mild dysfunction (68.3 +/- 7.2 mm vs 55.0 +/- 4.2 mm, p = 0.046), follow up left ventricular end-diastolic dimension, severe versus mild (68.7 +/- 4.1 mm vs 52.0 +/- 5.7 mm, p = 0.002), and follow-up fractional shortening, severe versus mild (14.6% +/- 5.0% vs 28.5% +/- 9.2%, p = 0.02) are significant. Six of the seven women with severe dysfunction had stable disease in follow-up and one is awaiting heart transplant. One of the two women with mild dysfunction had disease resolution and one had stable disease. CONCLUSION: Patients with severe myocardial dysfunction due to peripartum cardiomyopathy are unlikely to regain normal cardiac function on follow-up. PMID- 9396908 TI - A randomized trial of epidural anesthesia to improve external cephalic version success. AB - OBJECTIVE: This study was designed to determine whether epidural anesthesia would improve external cephalic version success in a safe and effective manner. STUDY DESIGN: All women > 37 weeks' gestation with breech presentation scheduled for external cephalic version at the medical center from Dec. 1, 1993, to July 31, 1996, were randomized to receive an epidural or no epidural anesthesia. Under ultrasonographic guidance up to three version attempts were performed. RESULTS: Sixty-nine women were randomized to receive epidural (n = 35) versus no epidural (n = 34) anesthesia for external cephalic version. There were no statistically significant differences in maternal age, parity, maternal weight, gestational age, estimated fetal weight, or station of the presenting part. The success rate was better for the epidural group (relative risk 2.12, 95% confidence interval 1.24 to 3.62). Neither anterior placentation or oligohydramnios affected the success rate. CONCLUSION: Epidural anesthesia increases success of external cephalic version without any apparent detrimental effect on the maternal-fetal unit. PMID- 9396909 TI - Impact of multiple antenatal doses of betamethasone on growth and development of mice offspring. AB - OBJECTIVE: Our purpose was to determine in a randomized, placebo-controlled manner whether multiple antenatal doses of betamethasone affect long-term growth and development of exposed mouse offspring. STUDY DESIGN: Sixty pregnant CD-1 mice received either two, four, or eight antepartum doses of 0.1 mg betamethasone or placebo. Perinatal outcomes, growth, and development of the offspring were compared in a blinded manner. Variables were compared by analysis of variance or chi 2 testing. RESULTS: Betamethasone-exposed subjects gained less weight during pregnancy and were delivered of fewer live pups, with fewer male survivors and lower birth weights. These trends were dose related. Growth measurements were similar after the neonatal period. No differences in functional development and physical maturation in the offspring were noted. The reproductive capability, perinatal outcomes, and growth and development of the second-generation offspring were unaffected by betamethasone exposure. CONCLUSION: Multiple antenatal dosings of betamethasone, reaching toxic levels, did not have an impact on the long-term growth and development of the surviving mouse offspring. PMID- 9396910 TI - Induction of labor with use of a Foley catheter and extraamniotic corticosteroids. AB - OBJECTIVE: Our purpose was to examine the hypothesis that corticosteroids, when administered extraamniotically, can enhance the labor process and reduce the induction-to-delivery interval. STUDY DESIGN: A double-blind, randomized study was conducted on 98 women with a gestational age of 36 to 42 weeks, an unfavorable cervix, and medical indications for delivery, who were assigned to receive either 20 mg of dexamethasone in saline solution (study group, n = 50) or saline solution only (control group, n = 48) administered extraamniotically through an intracervical inflated Foley balloon catheter. The net effect of steroids was assessed with use of multivariant logistic regression analysis. RESULTS: The mean time intervals between induction of labor to the active phase and between induction of labor to delivery were significantly shorter in the study group compared with those of the control group (7.8 +/- 3.1 hours vs 9.9 +/ 3.9 hours, p < 0.03, 11.9 +/- 3.0 hours vs 14.5 +/- 4.8 hours, p < 0.01, respectively). Those not receiving steroids were at a 3.2 higher risk of having a longer time interval of induction to delivery (95% confidence interval 1.1 to 9.5). The general success rate in achieving vaginal delivery was, however, similar between the groups. CONCLUSIONS: Induction of labor with use of an intracervical Foley balloon catheter and extraamniotic corticosteroids reduces the time interval from induction of labor to delivery. This may indicate a possible role for corticosteroids in the parturition process. PMID- 9396911 TI - Identifying twin gestations at low risk for preterm birth with a transvaginal ultrasonographic cervical measurement at 24 to 26 weeks' gestation. AB - OBJECTIVE: Because twins are a high-risk group for preterm birth, many clinicians routinely use prophylactic interventions such as home bed rest, hospital bed rest, oral tocolytics, or home uterine activity monitoring to prevent preterm delivery. We sought to identify twin gestations at low risk for spontaneous preterm birth with transvaginal ultrasonography of the cervix to avoid the unnecessary use of prophylactic interventions in these pregnancies. STUDY DESIGN: We measured cervical length at 24 to 26 weeks' gestation by transvaginal ultrasonography in women with twin gestations referred to our prematurity prevention clinic. Each delivery was classified as (1) spontaneous preterm birth < 34 weeks' gestation, (2) delivery at > or = 34 weeks' gestation with intervention, or (3) delivery at > or = 34 weeks' gestation without intervention. Intervention included strict bed rest at home or in the hospital, either parenteral or oral tocolysis, or both, or home uterine activity monitoring. Indicated preterm deliveries and patients with cerclage were excluded from this analysis. The ability of transvaginal cervical length to predict women who would deliver at > or = 34 weeks without intervention was evaluated. A cervical length of 35 mm was chosen by scatter diagram as the best cutoff to discriminate between the group delivered at term without intervention and the other two groups. RESULTS: Of 85 women with twin gestations who underwent ultrasonographic cervical length measurements at 24 to 26 weeks' gestation, 17 had spontaneous preterm birth at < 34 weeks, 23 were delivered at > or = 34 weeks but required intervention, and 45 were delivered at > or = 34 weeks without intervention. The mean cervical length for those delivered at > or = 34 weeks' gestation without intervention (36.4 +/- 5.8 mm) was significantly greater (p < 0.0001) than the mean for those delivered preterm (27.4 +/- 8.5) and those delivered at > or = 34 weeks' gestation who required intervention (27.7 +/- 10.5 mm). The sensitivity, specificity, and positive and negative predictive values of a cervical length > 35 mm for predicting delivery at > or = 34 weeks' gestation are 49%, 94%, 97%, and 31%, respectively. CONCLUSION: A transvaginal ultrasonographic measurement of the cervix of > 35 mm at 24 to 26 weeks in twin gestations can identify patients who are at low risk for delivery before 34 weeks' gestation. PMID- 9396912 TI - The challenge of complementary and alternative medicine. AB - Complementary and alternative medicine can be defined as those medical systems, practices, interventions, and applications that currently are not part of the dominant or conventional medical system. There are more than 300 different topics under the term complementary and alternative medicine that can be divided into seven major categories on the basis of philosophy, approach to the patient, and orientation. Most patients seeking care from complementary and alternative medicine providers do so for the relief of signs and symptoms related to chronic illness while they are under the care of a physician. Clinical data derived from appropriately conducted clinical trials support the use and value of complementary and alternative medicine for selected indications. The challenge for both conventional medicine and complementary and alternative medicine is to fulfill the role of patient advocate by engaging in reciprocal open communication, facilitating the patient's informed choice, avoiding harmful or useless practices, and implementing an integrated evidence-based care plan. PMID- 9396913 TI - Brachial plexus palsy associated with cesarean section: an in utero injury? AB - OBJECTIVE: Brachial plexus injury may be unrelated to manipulations performed at the time of delivery, occurring in the absence of shoulder dystocia and in the posterior arm of infants with anterior shoulder dystocia. To further support the hypothesis that some of these nerve injuries appear to be of intrauterine origin, we present a series of brachial plexus palsies associated with atraumatic cesarean delivery among fetuses presenting in the vertex position. STUDY DESIGN: We performed a computerized search of all deliveries from 1991 to 1995 for the discharge diagnoses of brachial plexus injury and cesarean section. Inclusion criteria included cephalic presentation at the time of delivery and the absence of traumatic delivery. RESULTS: We noted six cases of Erb's palsy, with four palsies in the anterior shoulder and two in the posterior arm. Among those five patients undergoing cesarean section because of labor abnormalities, two had uterine cavity abnormalities whereas one had a prolonged second stage of labor. One brachial plexus palsy occurred in the absence of active labor. All nerve injuries were persistent at age 1 year. CONCLUSIONS: Brachial plexus palsy can be associated with cesarean delivery. Such palsies appear to be of intrauterine origin and are more likely to persist. PMID- 9396914 TI - Congenital malformations in offspring of women with hyperglycemia first detected during pregnancy. AB - OBJECTIVES: Our aim was to determine risk factors for congenital malformations in offspring of women with hyperglycemia first detected during pregnancy (i.e., women with gestational diabetes). STUDY DESIGN: A total of 3743 pregnancies complicated by gestational diabetes mellitus delivered at > 20 weeks of gestation were reviewed for the presence of congenital malformations diagnosed before hospital discharge. Anomalies were categorized as major, minor, or absent. Pregnancies with genetic syndromes and aneuploidies were excluded. In addition to maternal clinical and historic parameters, diagnostic glycemic parameters (fasting and post-glucose-challenge levels from the diagnostic glucose tolerance test, highest fasting serum glucose level, and hemoglobin A1c level before insulin therapy) were examined by logistic regression for predictive risk of major anomalies. RESULTS: One or more major congenital anomalies were present in 108 (2.9%) of the newborns; an additional 91 (2.4%) had only minor anomalies. None of the maternal variables were associated with the risk of minor anomalies. By contrast, parity, a history of gestational diabetes mellitus, and several glycemic parameters were associated with the risk of major anomalies. The highest fasting serum glucose level was the best independent predictor (odds ratio 1.13/10 mg/dl, 95% confidence interval 1.09 to 1.34). The fasting serum glucose level at diagnosis, a parameter that is almost uniformly available to clinicians, gave similar predictive information about the risk of major anomalies (odds ratio 1.13, 95% confidence interval 1.08 to 1.14). Stratification of women into subgroups of fasting serum glucose level at diagnosis revealed the incidence of major anomalies to be as follows: 2.1% with a fasting serum glucose level < 120 mg/dl (2973 pregnancies), 5.2% with a fasting serum glucose level of 121 to 260 mg/dl (747 pregnancies), and 30.4% with a fasting serum glucose level > 260 mg/dl (23 pregnancies). CONCLUSION: In a large population of women without a diagnosis of diabetes before pregnancy, the maternal fasting serum glucose concentration at diagnosis was a useful predictor of the risk of major but not minor anomalies. The rate of major anomalies doubled with a fasting glucose level > 120 mg/dl. Thus a fasting glucose level below that of overt diabetes outside of pregnancy carries an important risk of major anomalies that must be considered in the counseling and management of these patients. PMID- 9396915 TI - Fetal plasma iron and restoration of red blood cell mass after hemorrhage of the ovine fetus. AB - OBJECTIVE: Our purpose was to determine whether the restoration of fetal red blood cell mass after acute hemorrhage of 40% of the fetal blood volume is related to fetal plasma iron concentration. STUDY DESIGN: Ten chronically catheterized ovine fetuses were monitored for 10 days beginning at 125 +/- 1 (SE) days of gestation. After a 3-day control period 40% of the fetal blood was removed over 2 hours at a rate of approximately 1 ml/min. Fetal plasma iron and erythropoietin concentrations, hematocrit, blood volume, and red blood cell mass were measured daily before and for 7 days after fetal hemorrhage. Statistical analysis was by analysis of variance, correlation, and regression. RESULTS: Although blood volume was restored within 3 days of the hemorrhage (101.0% +/- 1.4% of prehemorrhage volume), red blood cell mass was not (81.8% +/- 2.8%). Only 6 of 10 fetuses restored their red blood cell mass to prehemorrhage levels by the end of the 7-day posthemorrhage period. On day 10 red blood cell mass correlated positively with prehemorrhage (r = 0.74, p = 0.015) and posthemorrhage (r = 0.69, p = 0.045) plasma iron concentration and negatively with posthemorrhage erythropoietin concentration (r = -0.68, p = 0.047). CONCLUSION: Fetal plasma iron concentration is an important factor in restoration of fetal red blood cell mass after loss of blood. The negative correlation of erythropoietin concentration with posthemorrhagic red blood cell mass suggests that iron, not erythropoietin, may be the limiting factor in recovery from hemorrhage-induced anemia. Thus iron supplementation of the fetus may be of benefit in the treatment of some types of fetal anemia. PMID- 9396916 TI - Is bleeding a predictor of endometrial hyperplasia in postmenopausal women receiving hormone replacement therapy? Menopause Study Group (United States, Italy, Netherlands, Switzerland, Belgium, Germany, and Finland. AB - OBJECTIVE: We evaluated bleeding as a predictor of endometrial hyperplasia in postmenopausal women receiving conjugated estrogens (Premarin) alone or with medroxyprogesterone acetate. STUDY DESIGN: This study was a retrospective assessment of bleeding and endometrial histologic data gathered during the prospective Menopause Study Group trial. RESULTS: Approximately 20% of women (n = 57) who received unopposed conjugated estrogens for 1 year had endometrial hyperplasia. These women (n = 56) had more bleeding days than did women who did not have hyperplasia (p < 0.001). For users of unopposed conjugated estrogens, the predictive value of amenorrhea to indicate a nonhyperplasia diagnosis was 95%. Irregular bleeding did not appear to be indicative of hyperplasia when continuous or cyclic medroxyprogesterone acetate was added to conjugated estrogens. CONCLUSION: Unanticipated prolonged bleeding in postmenopausal women receiving unopposed estrogen is suggestive of endometrial hyperplasia. Some irregular bleeding is anticipated in women who receive conjugated estrogens plus medroxyprogesterone acetate as they acclimate to therapy, and this was not associated with hyperplasia. PMID- 9396917 TI - Bacterial vaginosis as a risk factor for upper genital tract infection. AB - OBJECTIVE: The objective of this study was to determine whether the clinical diagnosis of bacterial vaginosis is associated with objective evidence of acute upper genital tract infection. STUDY DESIGN: Women who either met the Centers for Disease Control and Prevention's minimal criteria for acute pelvic inflammatory disease or had other "nonclassic" signs of upper genital tract infection (i.e., atypical pelvic pain, abnormal uterine bleeding, or cervicitis) were evaluated with either an endometrial biopsy or a laparoscopy with endometrial and fimbrial biopsies for objective evidence of upper genital tract infection. Bacterial vaginosis was considered present if three of the four following criteria were found: (1) homogeneous gray-white vaginal discharge, (2) vaginal pH > 4.5, (3) positive "whiff" test result, and (4) the presence of > 20% of epithelial cells classified as clue cells. Patients were considered to have upper genital tract infection if they had histologic, microbiologic, or laparoscopic evidence of upper tract infection. RESULTS: One hundred sixteen women were evaluated between August 1993 and March 1997 with complete evaluations. Objective evidence of upper tract infection was present in 56% (14/25) of women with the clinical diagnosis of bacterial vaginosis compared with 30% of women (27/91) who did not meet the clinical criteria (p = 0.015). Using logistic regression to control for confounding variables, we found that the presence of bacterial vaginosis was associated with a threefold increased risk of upper genital tract infection (adjusted odds ratio = 3.0, 95% confidence interval 1.2 to 7.6). CONCLUSIONS: Bacterial vaginosis is associated with an increased risk of objective evidence of acute upper genital tract infection. Future prospective studies are needed to determine whether treatment of bacterial vaginosis can reduce the risk of ascending infection. PMID- 9396918 TI - Biopsy findings in five hundred thirty-one patients with atypical glandular cells of uncertain significance as defined by the Bethesda system. AB - OBJECTIVES: Histologic findings in biopsy specimens obtained from patients with atypical glandular cells of uncertain significance were studied to define the utility and limitations of this category. STUDY DESIGN: Computerized records over a 3-year period were retrospectively analyzed. The most significant histologic diagnosis from all biopsy specimens submitted was compared with the subcategory of the first Papanicolaou smear obtained showing atypical glandular cells of uncertain significance. RESULTS: Biopsy results were available for 531 of 1117 patients with atypical glandular cells of uncertain significance (48%). Biopsy proved preinvasive (83%) or invasive (17%) lesions were present in 191 patients (36%). Eighty-nine percent of the preinvasive lesions were squamous, whereas 97% of the invasive lesions were glandular. Glandular lesions were more likely to be invasive, whereas squamous lesions were more likely to be preinvasive (p < 0.001). Twenty-eight patients had endometrial carcinoma, which represents 88% of all invasive carcinomas detected. CONCLUSIONS: Almost three fourths of patients with atypical glandular cells of uncertain significance and with lesions have squamous lesions, not glandular as suggested by the name of the category. Unlike patients with atypical squamous cells of uncertain significance, patients with atypical glandular cells of uncertain significance have a significant risk of malignant lesions, which are nearly all glandular and predominantly arise from the endometrium. PMID- 9396920 TI - Heart rate and blood pressure variabilities are increased in pregnancy-induced hypertension. AB - OBJECTIVE: Our purpose was to study whether cardiovascular changes in pregnancy induced hypertension are associated with the increase in sympathetic control of hemodynamics and change in sympathovagal balance. STUDY DESIGN: Fourteen women with pregnancy-induced hypertension and 16 women with uncomplicated pregnancies of similar duration were studied. Electrocardiographic signals and arterial blood pressure (Finapres monitor, Ohmeda) were continuously measured noninvasively throughout the study. Heart rate and blood pressure were measured while the subject was breathing (1) with her normal tidal volume at a frequency of 15 breaths per minute and (2) as deeply as possible at a frequency of six breaths per minute. Heart rate and systolic blood pressure variability were calculated with use of the autoregressive model of spectral analysis. RESULTS: Heart rate and systolic blood pressure variabilities were significantly increased in women with pregnancy-induced hypertension compared with normotensive pregnant women. This increase was greatest in the high frequency component of heart rate variability (p = 0.02) while the women were breathing with a normal tidal volume. Further, the medium frequency (p = 0.03) and high-frequency variabilities (p = 0.03) of systolic blood pressure were significantly increased in women with preeclampsia compared with normotensive pregnant subjects. CONCLUSIONS: Neural control of the heart rate and blood pressure are disturbed in pregnancy-induced hypertension, as shown by increased heart rate and blood pressure variability. Both the sympathetic and parasympathetic control of the heart rate and blood pressure appear to be increased. The maladaptation of the cardiovascular system in women with pregnancy-induced hypertension is manifested as a lack of the physiologic decline in cardiovascular oscillations. PMID- 9396919 TI - Strategies to reduce the incidence of endometrial cancer in postmenopausal women. AB - OBJECTIVE: The purpose of this study was to develop strategies to reduce the incidence of endometrial cancer in postmenopausal women by reviewing methods to diagnose women at risk and providing therapies that are effective. STUDY DESIGN: A literature search for studies between 1975 and 1995 of the association between hormone therapy and endometrial cancer was performed with MEDLINE. There were nine reports with a total of 66 cases of endometrial cancer developing during use of estrogen-progestogen replacement therapy. RESULTS: The cases of endometrial cancer diagnosed during use of estrogen-progestogen therapy because of abnormal bleeding occurred when the dosage or duration of the progestogen was less than that considered optimum. The minimum effective dosage of medroxyprogesterone acetate is 10 mg and the minimum effective dosage of norethindrone acetate to 2.5 mg. The minimum effective duration of the progestogen is 12 to 14 days when it is used sequentially. Continuous combined therapy with low dosage estrogen and progestogen may not be fully endometrial protective. CONCLUSIONS: The progestogen challenge test is an effective test to identify women at increased risk for endometrial cancer. There are no adverse effects on lipids and lipoproteins from added progestogen when adequate dosages of estrogen are given. Side effects of progestogens can be managed with mild diuretics or by changing the type, dosage, or route of administration. Cyclic combined therapy may be more endometrial protective than continuous combined hormone replacement is. PMID- 9396921 TI - Persistent cerebrovascular changes in postpartum preeclamptic women: a Doppler evaluation. AB - OBJECTIVE: Our goal was to evaluate the cerebral vasculature in postpartum normotensive and preeclamptic women. STUDY DESIGN: Nineteen previously preeclamptic women and 19 matched normotensive controls were studied at 6 weeks, and 8 preeclamptic women and 28 normotensive controls were studied at 12 weeks post partum. Systolic, diastolic, and mean velocities, as well as resistance and pulsatility indexes, of the middle cerebral, ophthalmic, and central retinal arteries were recorded. Data are presented as median and range. Statistical significance was set at p < 0.05. RESULTS: There were no differences in maternal age, parity, heart rate, mean arterial pressure, and proteinuria between the two groups at 6 and 12 weeks post partum. At 6 weeks post partum the preeclamptic group had higher ophthalmic artery diastolic velocity (9.0 cm/sec, 3.1 to 22.3, vs 5.4 cm/sec, 3.0 to 20.1; p = 0.008), ophthalmic artery mean velocity (6.0 cm/sec, 8.8 to 34.8, vs 12.5 cm/sec, 6.8 to 35.4; p = 0.03), and central retinal artery systolic velocity (10.0 cm/sec, 7.6 to 28.0, vs 8.4 cm/sec, 5.2 to 18.3; p = 0.02). The ophthalmic artery resistance index (0.72, 0.43 to 0.88, vs 0.79, 0.66 to 0.90; p = 0.03) and ophthalmic artery pulsatility index (1.56, 0.94 to 2.82, vs 2.03, vs 1.13 to 3.10; p = 0.04) were lower in the preeclamptic group. At 12 weeks post partum the preeclamptic group had elevated ophthalmic artery mean velocity (14.5 cm/sec, 7.9 to 20.2, vs 10.9 cm/sec, 5.5 to 15.4 p = 0.01), central retinal artery systolic velocity (11.1 cm/sec, 6.8 to 15.9, vs 8.5, 5.1 to 15.3; p = 0.02), and central retinal artery diastolic velocity (3.9 cm/sec, 1.2 to 5.2, vs 3.0, 1.4 to 5.8; p < 0.05). CONCLUSION: In the postpartum period preeclamptic women show persistently elevated central retinal artery systolic velocity, which suggests distal vasoconstriction. PMID- 9396922 TI - The role of deoxyribonucleic acid image cytometric and interphase cytogenetic analyses in the differential diagnosis, prognosis, and clinical follow-up of hydatidiform moles. A report from the Central Molar Registration in The Netherlands. AB - OBJECTIVES: To assess the value of deoxyribonucleic acid ploidy in the differential diagnosis and clinical follow-up of hydatidiform moles, the histopathologic features, deoxyribonucleic acid ploidy, and clinical follow-up were compared in 347 cases: 143 complete moles, 52 partial moles, and 152 abortions, of which 56 cases were hydropic abortions with histologic features of triploidy but lacked trophoblastic hyperplasia. STUDY DESIGN: In all cases deoxyribonucleic acid image cytometry was performed, and in 85 of these cases interphase cytogenetics was also performed. RESULTS: With use of deoxyribonucleic acid image cytometry and interphase cytogenetics, a bimodal polyploid deoxyribonucleic acid pattern was present in 97% of complete moles, 27% of partial moles, and 4% of abortions. All these cases of partial mole were reclassified to complete mole on the basis of this deoxyribonucleic acid pattern and the histopathologic features in spite of the presence of fetal blood cells, amnion, or yolk sac. Deoxyribonucleic acid triploidy was found in 95% of the remaining partial moles, in 77% of hydropic abortions with histologic features of triploidy, and in 14% of the remaining abortions. Reliable differentiation between deoxyribonucleic acid triploid partial moles and hydropic abortions with histologic features of triploidy was not possible on basis of the histopathologic features (trophoblastic hyperplasia) or 3.5c exceeding rates. Deoxyribonucleic acid diploidy was found in 1% of complete moles, 23% of hydropic abortions with features of triploidy, and 78% of the remaining abortions. Deoxyribonucleic acid tetraploidy was rarely found (1% of complete moles, 2% of partial moles, 1% of abortions). Persistent gestational trophoblastic disease developed in 33% of the bimodal deoxyribonucleic acid polyploid cases (all complete moles), in 1% of the diploid cases (concerning one of the two diploid complete moles), and in 1% of the triploid cases (partial moles). CONCLUSION: Deoxyribonucleic acid analysis is essential in the diagnosis of hydatidiform moles to decide on clinical follow-up. PMID- 9396923 TI - Lipopolysaccharide binding protein and soluble CD14 receptor protein in amniotic fluid and cord blood in patients at term. AB - OBJECTIVES: Our purpose was to examine whether lipopolysaccharide binding protein and soluble CD14 are present in amniotic fluid and to determine whether the lipopolysaccharide binding protein and soluble CD14 concentrations are associated with indicators of infection or labor at term. A lipopolysaccharide lipopolysaccharide binding protein complex activates macrophages through soluble CD14 at lipopolysaccharide concentrations up to 100 times lower than required with lipopolysaccharide alone. Thus lipopolysaccharide binding protein and soluble CD14 in amniotic fluid could explain the high concentrations of cytokines found in amniotic fluid of culture-positive patients and may even explain the presence of cytokines in some culture-negative patients. STUDY DESIGN: Healthy women at term undergoing cesarean section had amniotic fluid, chorioamnion, decidua, and cord blood obtained. Lipopolysaccharide binding protein was measured by enzyme-linked immunosorbent assay. Amniotic fluid was cultured and assayed for cytokines, and the chorioamnion and decidua were cultured and examined histologically. RESULTS: Lipopolysaccharide binding protein and soluble CD14 were present in all amniotic fluids and fetal cord blood. An elevated level of lipopolysaccharide binding protein (270 ng/ml/mg of protein) was present in the amniotic fluid of 12 (36%) of the 33 patients. An elevated level was associated with microorganisms in the chorioamnion and decidua, cytokines (tumor necrosis factor-alpha, interleukin-6, and interleukin-8) in amniotic fluid, histologic chorioamnionitis, and labor. Among patients in labor, the concentration of lipopolysaccharide binding protein appeared independent of microorganisms in the amniotic fluid. CONCLUSIONS: Lipopolysaccharide binding protein and soluble CD14 are present in amniotic fluid, and concentrations of lipopolysaccharide binding protein are elevated in patients in labor with and without evidence of infection. Lipopolysaccharide binding protein and soluble CD14 may mediate intrauterine inflammatory responses at term. PMID- 9396924 TI - Multicenter study on the clinical value of fetal pulse oximetry. I. Methodologic evaluation. The French Study Group on Fetal Pulse Oximetry. AB - OBJECTIVE: Our purpose was to evaluate the feasibility of intrapartum fetal pulse oximetry, the distribution of fetal oxygen saturation values, and the relationship with the neonatal outcome in a population with an abnormal fetal heart rate. STUDY DESIGN: A prospective multicenter observational study was performed from June 1994 to November 1995. Fetal oxygen saturation was continuously recorded with use of a Nellcor N-400 fetal pulse oximeter in case of an abnormal fetal heart rate during labor. Simultaneous readings of fetal oxygen saturation and fetal blood analysis were obtained at inclusion and before birth. Feasibility, adverse effects, distribution of fetal oxygen saturation values, and relationship with neonatal outcome were assessed. RESULTS: One hundred seventy four patients were included. From 172 attempted sensor placements, the procedure was impossible in three cases and fetal oxygen saturation values were obtained in 164 cases (95.3%). Physicians considered sensor placement an easier task than an attempt at fetal blood analysis (easy in 87.5% vs 78.9% for fetal blood analysis, p = 0.03). The mean reliable signal time (+/- SD) was 64.7% +/- 32% during the first stage. There were no serious adverse effects in the study population. The mean fetal oxygen saturation during the first stage of labor was 42.2% +/- 8.0% (10th to 90th percentile range 30% to 53%). Fetal oxygen saturation was significantly correlated with scalp pH (r = 0.29, p = 0.01) but not with neonatal umbilical artery pH or gas values. There was a significant association between low fetal oxygen saturation (< 30%) and poor neonatal condition. CONCLUSION: The feasibility of fetal pulse oximetry is satisfactory in clinical practice. It is easy to use and provides a fair rate of recorded values, even in a population with suspicion of fetal distress. A low fetal oxygen saturation is significantly associated with an abnormal neonatal outcome. PMID- 9396925 TI - Circulating bioactive tumor necrosis factor-alpha, tumor necrosis factor-alpha receptors, fibronectin, and tumor necrosis factor-alpha inducible cell adhesion molecule VCAM-1 in uncomplicated pregnancy. AB - OBJECTIVES: Our goal was to assess in a longitudinal study of uncomplicated pregnancy the course of maternal plasma concentrations of the bioactive cytokine tumor necrosis factor-alpha, the soluble tumor necrosis factor-alpha receptors sTNFRI and sTNFRII, the soluble cell adhesion molecule sVCAM-1, and circulating fibronectin. STUDY DESIGN: Blood was collected from 22 healthy pregnant women at 7 to 17, 18 to 22, 23 to 28, and 30 to 36 weeks' gestation and post partum. Plasma samples were measured by bioassay for bioactive tumor necrosis factor alpha, by immunoassay for sTNFRI, sTNFRII, and VCAM-1, and by radial immunodiffusion for circulating fibronectin, and data were statistically analyzed. RESULTS: Plasma concentrations of all variables were significantly linked with gestational age. Levels of bioactive tumor necrosis factor-alpha and sTNFRII showed a parallel rise in the second trimester and a decrease thereafter. Values for sTNFRI and sTNFRII and for these receptors and VCAM-1 were correlated, a weak correlation between bioactive tumor necrosis factor-alpha and sTNFRII was observed, and no correlation between circulating fibronectin and other variables was apparent. CONCLUSIONS: All variables studied exhibited a characteristic pattern depending on gestational age, which supports the concept of a physiologic role of tumor necrosis factor-alpha in pregnancy. PMID- 9396926 TI - Coordinate expression of inducible nitric oxide synthase and cyclooxygenase-2 genes in uterine tissues of endotoxin-treated pregnant mice. AB - OBJECTIVES: Our purpose was to investigate the relationship between expression of cyclooxygenase-2 and inducible nitric oxide synthase genes after labor induction with bacterial lipopolysaccharide in a murine model of preterm parturition. STUDY DESIGN: Pregnant C57B1/6 mice were given Escherichia coli lipopolysaccharide (20 micrograms per mouse) by intraperitoneal injection on day 16 of gestation, and the animals were followed up for signs of labor. Control mice received an equivalent volume of 0.9% saline solution. The latency from lipopolysaccharide injections until appearance of the first pup was recorded. Two separate groups of mice were given either aminoguanidine or indomethacin (5 mg/kg intragastric) 24 hours before induction of preterm labor. In a separate set of experiments mice were treated with lipopolysaccharide as described and were killed at intervals from 0.5 to 72 hours and intrauterine tissues (uterus, placenta, and fetal membranes) were removed and snap frozen in liquid nitrogen. Total protein and ribonucleic acid were extracted for Western and Northern blot analysis of cyclooxygenase-2 and inducible nitric oxide synthase protein and messenger ribonucleic acid, respectively. RESULTS: Northern blots from uterine, placental, and fetal membrane tissues of lipopolysaccharide- and saline solution-treated mice revealed that cyclooxygenase-2 and inducible nitric oxide synthase messenger ribonucleic acid transcripts were rapidly (within 0.5 to 2 hours) up-regulated after lipopolysaccharide administration but were unchanged in mice injected with saline solution. Immunoblot analysis with isoform-specific antibodies revealed that both enzymes were expressed in uterus, placenta, and fetal membranes in a coordinated fashion with peak expression seen at 6 to 8 hours. Although the steady-state accumulation of messenger ribonucleic acid transcripts encoding cyclooxygenase-2 and inducible nitric oxide synthase peaked at 6 hours and declined to baseline by 16 hours after injection with lipopolysaccharide, expression of cyclooxygenase-2 and inducible nitric oxide synthase was sustained through the period when premature delivery was observed. Nitric oxide-dependent cyclooxygenase-2 and inducible nitric oxide synthase expression was demonstrated by the elimination of accumulation of both messenger ribonucleic acid transcripts in mice pretreated with aminoguanidine before injection with lipopolysaccharide. CONCLUSIONS: These data indicate that nitric oxide synthesis may be a prerequisite for subsequent stimulation of cyclooxygenase-2 and inducible nitric oxide synthase gene expression. Taken together, the data suggest that cyclooxygenase-2 and inducible nitric oxide synthase are expressed in a coordinated manner in the uterus of endotoxin-challenged pregnant mice and that their enzymatic products may contribute to the signaling of uterine activity or cervical changes culminating in expulsion of the fetus. PMID- 9396927 TI - Role of nitric oxide in the regulation of vascular tone in pressurized and perfused resistance myometrial arteries from term pregnant women. AB - OBJECTIVE: Our purpose was to evaluate flow-induced responses, myogenic tone, and norepinephrine-induced constriction in myometrial resistance arteries from normal term pregnant women and the role that nitric oxide and prostanoids may play in these responses. STUDY DESIGN: Arteries (approximately 200 microns, n = 14, at 40 mm Hg) were dissected from myometrial biopsy specimens from women undergoing cesarean section and then were mounted in a pressure arteriograph. Responses to intraluminal flow, pressure, and a constrictor agonist (norepinephrine 10(-6) mol/L) were studied in the absence and presence of N omega-nitro-L-arginine methyl ester (n = 7) or indomethacin (n = 5). Myogenic and norepinephrine-induced tone were calculated after the determination of artery diameter in the absence of extracellular calcium. RESULTS: Arteries developed myogenic tone (80 mm Hg) that was not modulated by nitric oxide or prostanoid release, whereas norepinephrine induced tone was significantly enhanced by the nitric oxide inhibitor. An increase in intraluminal flow led to dilatation in physiologic salt solution and indomethacin, but to constriction in the presence of N omega-nitro-L-arginine methyl ester (percent increase in diameter at flow rate of 184.6 microliters/min, 24% +/- 8% in physiologic salt solution and 20% +/- 4% in the presence of indomethacin versus -27% +/- 12% in N omega-nitro-L-arginine methyl ester alone and -21% +/- 10% in indomethacin and N omega-nitro-L-arginine methyl ester, respectively, analysis of variance, p < 0.05). CONCLUSIONS: Flow-induced shear stress is a physiologic modulator of vascular tone in myometrial arteries from pregnant women. Nitric oxide, but not prostanoids, mediates this response and also blunts norepinephrine constriction. Nitric oxide may play a fundamental role in the maintenance of adequate blood supply to the fetus during human pregnancy. PMID- 9396928 TI - New technologies? PMID- 9396929 TI - Fetal sex and cesarean section. PMID- 9396930 TI - Public health approach to activated protein C resistance assay. PMID- 9396931 TI - Effect of hormone replacement therapy on aortic compliance in postmenopausal women. PMID- 9396932 TI - Flawed study alleging prevention of diabetic embryopathy. PMID- 9396933 TI - Intrahepatic cholestasis of pregnancy: are we really able to predict fetal outcome? PMID- 9396934 TI - Endometriosis and non-Hodgkin's lymphoma. PMID- 9396935 TI - Postmenopausal female hormone use and venous thromboembolic disease in black women. PMID- 9396936 TI - Cost of the treatment of unruptured ectopic pregnancy: is there still a place for laparotomy? PMID- 9396937 TI - Umbilical cord compression, persistent pulmonary hypertension, and placental villous hypervascularity. PMID- 9396938 TI - What shape are we in? Gender, psychopathology, and the brain. PMID- 9396939 TI - Linkage and association in complex genetic diseases. PMID- 9396940 TI - Psychopathology in women and men: focus on female hormones. AB - OBJECTIVE: The goal of this overview is to examine male/female differences in psychopathology in light of the known effects of gonadal steroids, especially estradiol, on neural function. METHOD: The epidemiology of specific psychopathological syndromes is highlighted with respect to male/female differences and discussed against the backdrop of recent neuroendocrine findings. RESULTS: A number of differences between the sexes in rates of illness and course of illness are documented, with Alzheimer's disease, schizophrenia, alcoholism, and mood and anxiety disorders each illustrating slightly different hormone mediated risks and buffers. CONCLUSIONS: Estrogens are neuroprotective with respect to neuronal degeneration, growth, and susceptibility to toxins. The cyclic fluctuations of estrogens and progesterone enhance the response to stress, which confers susceptibility to depression and anxiety. PMID- 9396941 TI - Sex differences in brain morphology in schizophrenia. AB - OBJECTIVE: The current literature on sex differences in schizophrenia with regard to structural brain abnormalities is inconsistent. Several studies have suggested that male and female patients may differ in severity of brain abnormalities. Efforts to explore this issue have been hindered by small study groups, unbalanced groups (i.e., those with many more men than women), or both. The relatively smaller number of female schizophrenic patients in most studies may have made it more difficult to detect differences between patients and comparison subjects. This study was designed to evaluate brain morphology in a carefully selected group of patients with schizophrenia and healthy comparison subjects who were balanced by sex. METHOD: Eighty patients (40 male and 40 female) and 80 healthy volunteers matched by sex and age were studied. Magnetic resonance imaging scans were analyzed with the use of an automated method that yields volumes of major brain regions. RESULTS: There was a significant sex-by-diagnosis interaction for ventricular volume, with male patients having significantly larger ventricles than male comparison subjects but female patients showing no significant enlargement in comparison with healthy female subjects. Although the overall distribution of structural brain differences was very similar in the male and female patients, the male patients had a greater number of significant abnormalities than the female patients. CONCLUSIONS: These findings indicate that male and female patients with schizophrenia have the same pattern of structural brain abnormalities, but male patients appear to manifest greater severity, especially with regard to ventricular enlargement. PMID- 9396942 TI - Sex-specific expression of Heschl's gyrus functional and structural abnormalities in paranoid schizophrenia. AB - OBJECTIVE: Evidence supports abnormal temporal lobe structure and function in schizophrenia. Some abnormalities, particularly involving the auditory cortex, appear to be sex specific. These findings were extended to anatomical and physiological descriptors. METHOD: The authors quantified the volume, surface area, and three-dimensional location of Heschl's gyri on magnetic resonance imaging (MRI) images of 21 patients with paranoid schizophrenia and 24 healthy comparison subjects. Neuromagnetic localizations of the 100-msec latency auditory evoked field (M100) were compared with MRI-determined locations of Heschl's gyri, computed as the geometric center of mass of the volume. RESULTS: Volumetric measures revealed small Heschl's gyri only in male patients. Asymmetry was found in the location of the Heschl's gyrus centroid (more anterior on the right) across all groups. Male comparison subjects had M100 locations posterior to the Heschl's gyrus centroid in the left hemisphere and close to the Heschl's gyrus centroid on the right, while male patients had M100 sources anterior to the Heschl's gyrus centroid on the left. All women had M100 locations posterior to the Heschl's gyrus centroid on the left and anterior to it on the right. CONCLUSIONS: These results demonstrate that some temporal lobe abnormalities in schizophrenia are sex specific. They also suggest that the anomalous lateralization of the auditory evoked field cannot be explained by a shift in the underlying anatomy, since the anatomical substrate is lateralized in both comparison subjects and patients of both sexes. These findings may indicate a sex specific functional reorganization in the auditory cortex in schizophrenia. PMID- 9396943 TI - Quantitative morphology of the cerebellum and fourth ventricle in childhood-onset schizophrenia. AB - OBJECTIVE: Studies have suggested that the maldeveloped neural circuitry producing schizophrenic symptoms may include the cerebellum. The authors found further support for this hypothesis by examining cerebellar morphology in severely ill children and adolescents with childhood-onset schizophrenia. METHOD: Anatomic brain scans were acquired with a 1.5-T magnetic resonance imaging scanner for 24 patients (mean age = 14.1 years, SD = 2.2) with onset of schizophrenia by age 12 (mean age at onset = 10.0 years, SD = 1.9) and 52 healthy children. Volumes of the vermis, inferior posterior lobe, fourth ventricle, and total cerebellum and the midsagittal area of the vermis were measured manually. RESULTS: After adjustment for total cerebral volume, the volume of the vermis and the midsagittal area and volume of the inferior posterior lobe remained significantly smaller in the schizophrenic patients. There was no group difference in total cerebellar or fourth ventricle volume. CONCLUSIONS: These findings are consistent with observations of small vermal size in adult schizophrenia and provide further support for abnormal cerebellar function in childhood- and adult-onset schizophrenia. PMID- 9396944 TI - Anterior cingulate gyrus dysfunction and selective attention deficits in schizophrenia: [15O]H2O PET study during single-trial Stroop task performance. AB - OBJECTIVE: Attentional deficits are a prominent aspect of cognitive dysfunction in schizophrenia. The anterior cingulate gyrus is proposed to be an important component of frontal attentional control systems. Structural and functional abnormalities have been reported in this region in schizophrenia, but their relationship to attentional deficits is unknown. The authors investigated the function of the anterior cingulate gyrus and the related neural systems that are associated with selective attention in patients with schizophrenia. METHOD: While subjects performed multiple blocks of a single-trial Stroop task, [15O]H2O positron emission tomography scans were obtained. Fourteen patients with schizophrenia were compared with 15 normal subjects matched for age, gender, and parental education. RESULTS: The patients with schizophrenia responded at the same rate but made more errors in color naming during the color-incongruent condition. Consistent with the authors' hypothesis, patients with schizophrenia showed significantly less anterior cingulate gyrus activation while naming the color of color-incongruent stimuli. CONCLUSIONS: Patients with schizophrenia fail to activate the anterior cingulate gyrus during selective attention performance. This finding adds to the understanding of the functional significance of the structural and metabolic abnormalities in schizophrenia that have been previously reported in this region of the brain. PMID- 9396945 TI - Auditory hallucinations and the temporal cortical response to speech in schizophrenia: a functional magnetic resonance imaging study. AB - OBJECTIVE: The authors explored whether abnormal functional lateralization of temporal cortical language areas in schizophrenia was associated with a predisposition to auditory hallucinations and whether the auditory hallucinatory state would reduce the temporal cortical response to external speech. METHOD: Functional magnetic resonance imaging was used to measure the blood-oxygenation level-dependent signal induced by auditory perception of speech in three groups of male subjects: eight schizophrenic patients with a history of auditory hallucinations (trait-positive), none of whom was currently hallucinating; seven schizophrenic patients without such a history (trait-negative); and eight healthy volunteers. Seven schizophrenic patients were also examined while they were actually experiencing severe auditory verbal hallucinations and again after their hallucinations had diminished. RESULTS: Voxel-by-voxel comparison of the median power of subjects' responses to periodic external speech revealed that this measure was reduced in the left superior temporal gyrus but increased in the right middle temporal gyrus in the combined schizophrenic groups relative to the healthy comparison group. Comparison of the trait-positive and trait-negative patients revealed no clear difference in the power of temporal cortical activation. Comparison of patients when experiencing severe hallucinations and when hallucinations were mild revealed reduced responsivity of the temporal cortex, especially the right middle temporal gyrus, to external speech during the former state. CONCLUSIONS: These results suggest that schizophrenia is associated with a reduced left and increased right temporal cortical response to auditory perception of speech, with little distinction between patients who differ in their vulnerability to hallucinations. The auditory hallucinatory state is associated with reduced activity in temporal cortical regions that overlap with those that normally process external speech, possibly because of competition for common neurophysiological resources. PMID- 9396946 TI - Synaptic elimination, neurodevelopment, and the mechanism of hallucinated "voices" in schizophrenia. AB - OBJECTIVE: After peaking during childhood, synaptic density in the human frontal cortex declines by 30%-40% during adolescence because of progressive elimination of synaptic connections. The characteristic age at onset of schizophrenia--late adolescence and early adulthood--suggests that the disorder could arise from irregularities involving this neurodevelopmental process. METHOD: A computer simulation of a speech perception neural network was developed. Connections within the working memory component of the network were eliminated on the basis of a "Darwinian rule" in order to model loss of synapses. As a comparison, neuronal cell death, also postulated as being linked to both neurodevelopment and schizophrenia, was simulated. The authors determined whether these alterations at low levels could enhance perceptual capacity and at high levels produce spontaneous speech percepts that simulate hallucinated speech or "voices." RESULTS: Eliminating up to 65% of working memory connections improved perceptual ability; beyond that point, network performance declined and speech hallucinations emerged. Simulating excitotoxic neuronal loss at low levels also improved network performance, but in excess it did not produce hallucinations. CONCLUSIONS: The model demonstrates perceptual advantages of selective synaptic elimination as well as selective neuronal loss, suggesting a functional explanation for these aspects of neurodevelopment. The model predicts that psychosis arises from a pathological extension of one of these neurodevelopmental trends, namely, synaptic elimination. PMID- 9396947 TI - Posttraumatic stress disorder and functioning and quality of life outcomes in a nationally representative sample of male Vietnam veterans. AB - OBJECTIVE: Although posttraumatic stress disorder (PTSD) is a highly prevalent and often chronic condition, the relationship between PTSD and functioning and quality of life remains incompletely understood. METHOD: The authors undertook an archival analysis of data from the National Vietnam Veterans Readjustment Study. The study subjects consisted of the nationally representative sample of male Vietnam veterans who participated in the National Vietnam Veterans Readjustment Study. The authors estimated PTSD at the time of the interview with the Mississippi Scale for Combat-Related Posttraumatic Stress Disorder. They examined the following outcomes: diminished well-being, physical limitations, bed day in the past 2 weeks, compromised physical health status, currently not working, and perpetration of violence. Logistic models were used to determine the association between PTSD and outcome; adjustment was made for demographic characteristics and comorbid psychiatric and other medical conditions. RESULTS: The risks of poorer outcome were significantly higher in subjects with PTSD than in subjects without PTSD in five of the six domains. For the outcome domains of physical limitations, not working, compromised physical health, and diminished well-being, these significantly higher risks persisted even in the most conservative logistic models that removed the shared effects of comorbid psychiatric and other medical disorders. CONCLUSIONS: The suffering associated with combat related-PTSD extends beyond the signs and symptoms of the disorder to broader areas of functional and social morbidity. The significantly higher risk of impaired functioning and diminished quality of life uniquely attributable to PTSD suggests that PTSD may well be the core problem in this group of difficult to treat and multiply afflicted patients. PMID- 9396948 TI - One-year follow-up of survivors of a mass shooting. AB - OBJECTIVE: This report describes a 1-year follow-up study of survivors of a mass shooting incident. Acute-phase data from this incident were previously reported in this journal. METHOD: The Diagnostic Interview Schedule/Disaster Supplement was used to assess 136 survivors at 1-2 months and again a year later, with a 91% reinterview rate. RESULTS: In the acute postdisaster period, 28% of subjects met criteria for posttraumatic stress disorder (PTSD), and 18% of subjects qualified for another active psychiatric diagnosis. At follow-up, 24% of subjects reported a history of postdisaster PTSD (17% were currently symptomatic), and 12% another current psychiatric disorder. Half (54%) of all 46 individuals identified as having had PTSD at either interview were recovered at follow-up, and no index predictors of recovery were identified. There were no cases of delayed-onset PTSD (beyond 6 months). Considerable discrepancy in identified PTSD cases was apparent between index and follow-up. Inconsistency in reporting, rather than report of true delayed-onset, was responsible for all PTSD cases newly identified at 1 year. The majority of subjects with PTSD at index who were recovered at follow-up reported no history of postdisaster PTSD at follow-up, suggesting considerable influence of fading memory. CONCLUSIONS: This study's findings suggest that disaster research that conducts single interviews at index or a year later may overlook a significant portion of PTSD. The considerable diagnostic comorbidity found in this study was the one robust predictor of PTSD at any time after the disaster. Disaster survivors with a psychiatric history, especially depression, may be most vulnerable to developing PTSD and therefore may deserve special attention from disaster mental health workers. PMID- 9396949 TI - Objective documentation of child abuse and dissociation in 12 murderers with dissociative identity disorder. AB - OBJECTIVE: The skepticism regarding the existence of dissociative identity disorder as well as the abuse that engenders it persists for lack of objective documentation. This is doubly so for the disorder in murderers because of issues of suspected malingering. This article presents objective verification of both dissociative symptoms and severe abuse during childhood in a series of adult murderers with dissociative identity disorder. METHOD: This study consisted of a review of the clinical records of 11 men and one woman with DSM-IV-defined dissociative identity disorder who had committed murder. Data were gathered from medical, psychiatric, social service, school, military, and prison records and from records of interviews with subjects' family members and others. Handwriting samples were also examined. Data were analyzed qualitatively. RESULTS: Signs and symptoms of dissociative identity disorder in childhood and adulthood were corroborated independently and from several sources in all 12 cases; objective evidence of severe abuse was obtained in 11 cases. The subjects had amnesia for most of the abuse and underreported it. Marked changes in writing style and/or signatures were documented in 10 cases. CONCLUSIONS: This study establishes, once and for all, the linkage between early severe abuse and dissociative identity disorder. Further, the data demonstrate that the disorder can be distinguished from malingering and from other disorders. The study shows that it is possible, with great effort, to obtain objective evidence of both the symptoms of dissociative identity disorder and the abuse that engenders it. PMID- 9396950 TI - Seasonal variation in the occurrence of homicide in Finland. AB - OBJECTIVE: Although seasonal variation in impulsive aggression related to circannual rhythms of central serotonin neurotransmission is a topic of current interest, there is little firm knowledge on seasonality in the occurrence of homicide. Longitudinal studies on the seasonal rhythms of platelet imipramine binding and L-tryptophan levels have placed the circannual peaks around January and February and the nadirs around May and August. The aim of this study was to test the hypothesis that the number of homicides is the lowest during winter and the highest during spring and summer. A secondary hypothesis was that the seasonal variations in homicides and violent suicides are correlated. METHOD: The largest database on the monthly occurrence of homicide thus far (N = 4,553) was used in this study, in which the monthly occurrence of all murders and manslaughters in Finland during the years 1957-1995 was analyzed. RESULTS: During winter the homicide rate was 6% below the expected rate. Correspondingly, during summer there was a 6% elevation above the expected homicide rate, but no significant peak was observed in spring. There was a significant association between the monthly occurrence of homicides and violent suicides but not between homicides and nonviolent suicides. CONCLUSIONS: The results suggest that a seasonal variation in the occurrence of homicide exists. On the basis of current literature, it could be hypothesized that this seasonal variation and the correlation between the monthly occurrence of homicides and violent suicides are associated with the observed circannual rhythms of serotonin transmission. PMID- 9396951 TI - Characteristics of borderline personality disorder associated with suicidal behavior. AB - OBJECTIVE: This study examined the relationship between characteristics of borderline personality disorder and suicidal behavior. The authors hypothesized that a specific feature of borderline personality disorder, impulsivity, and childhood trauma, a possible etiological factor in the development of impulsivity, would be associated with suicidal behavior. METHOD: Information on lifetime history of suicidal behavior was obtained from 214 inpatients diagnosed with borderline personality disorder by structured clinical interview. The authors examined the relationship between DSM-III-R criteria met and the following measures of suicidal behavior: presence or absence of a previous suicide attempt, number of previous attempts, and lethality and intent to die associated with the most lethal lifetime attempt. RESULTS: Impulsivity was the only characteristic of borderline personality disorder (excluding the self destructive criterion) that was associated with a higher number of previous suicide attempts after control for lifetime diagnoses of depression and substance abuse. Global severity of pathology of borderline personality disorder was not associated with suicidal behavior. History of childhood abuse correlated significantly with number of lifetime suicide attempts. CONCLUSIONS: The trait of impulsivity is associated with number of lifetime suicide attempts and may therefore be a putative risk factor for a future suicide attempt. If so, impulsivity is a potential target therapeutically for prevention of future suicide attempts. The association between childhood abuse and number of lifetime suicide attempts is consistent with the hypothesis that childhood abuse is an etiological factor in the development of self-destructive behaviors. PMID- 9396952 TI - HIV seroprevalence among suicide victims in New York City, 1991-1993. AB - OBJECTIVE: The authors sought to determine the HIV seroprevalence among suicide victims in New York City. METHOD: All suicides of city residents from 1991 through 1993 were studied. The crude proportion of all suicide victims who were HIV positive and the proportion adjusted to the age, gender, and racial/ethnic characteristics of the New York City population were determined. The demographically adjusted proportion was then contrasted with HIV seroprevalence estimates for the New York City general population. HIV-seropositive suicide victims were assessed for pathological findings suggestive of HIV-related illnesses. RESULTS: The crude proportion of all suicide victims who were HIV seropositive was 0.088, and the demographically adjusted proportion was 0.049. Over 90% of all HIV-positive suicide victims were aged 25 to 54 years, and almost 90% were men. Among black and Hispanic men aged 35 to 54 years who committed suicide, the proportion who were HIV seropositive was 0.252--the highest seropositive rate of any demographic group. More than two-thirds of HIV seropositive suicide victims had no HIV-related pathology or AIDS-indicator conditions at autopsy. CONCLUSIONS: The demographically adjusted proportion of suicide victims who were HIV positive (approximately 0.038 to 0.059), contrasted with the HIV seroprevalence estimates for the New York City general population (approximately 0.014 to 0.032), the absence of HIV-related pathology among suicide victims, and the likelihood that many HIV-positive individuals had other risk factors for suicide, such as substance abuse, suggests that a positive HIV serostatus is associated, at most, with a modest elevation in suicide risk. PMID- 9396953 TI - Clinical and methodological factors related to reliability of the best-estimate diagnostic procedure. AB - OBJECTIVE: The reliability and accuracy of the best-estimate diagnostic procedure were examined, and factors associated with reliability were determined. METHOD: The subjects were 134 members of large multigenerational pedigrees densely affected by bipolar disorders or schizophrenia. Three best-estimate diagnoses were derived: first, by a research psychiatrist and research assistant unblind to the relatives' diagnoses; second, by two blind independent psychiatrists; third, by a panel of four blind psychiatrists. The subjects were characterized on several clinical and methodological variables, which were used to compare the agreements of two types of best-estimate diagnoses with the disagreements. RESULTS: There was satisfactory agreement between the unblind and blind consensus best-estimate diagnoses and between the two blind independent psychiatrists. Latent class analyses revealed that limited sensitivity was the main source of imperfect reliability. Confusability analyses revealed that the most problematic diagnostic distinctions involved schizoaffective disorder, which was confused with schizophrenia, bipolar I disorder, and schizophreniform disorder. Blindness significantly affected diagnostic outcome in latent class analyses. Moreover, for diagnostic disagreements, unblind diagnoses had greater continuity with the most predominant diagnosis in the pedigree than did blind diagnoses. Diagnostic disagreements were associated with the presence of mixed affective and psychotic symptoms, less diagnostic certainty, and shorter duration of illness. CONCLUSIONS: These results suggest that it is possible to identify cases that are more likely to lead to diagnostic disagreements in family and epidemiological studies and that blind diagnoses may help to prevent false positive diagnoses, which may be particularly detrimental to genetic linkage analyses. PMID- 9396954 TI - Mental disorders and disability among patients in a primary care group practice. AB - OBJECTIVE: This article examines social and occupational disability associated with several DSM-IV mental disorders in a group of adult primary care outpatients. METHOD: The subjects were 1,001 primary care patients (aged 18-70 years) in a large health maintenance organization. Data on each patient's sociodemographic characteristics and functional disability, including scores on the Sheehan Disability Scale, were collected at the time of a medical visit. A structured diagnostic interview for current DSM-IV disorders was then completed by a mental health professional over the telephone within 4 days of the visit. RESULTS: The most prevalent disorders were phobias (7.7%), major depressive disorder (7.3%), alcohol use disorders (5.2%), generalized anxiety disorder (3.7%), and panic disorder (3.0%). A total of 8.3% of the patients met the criteria for more than one mental disorder. The proportion of patients with co occurring mental disorders varied by index disorder from 50.0% (alcohol use disorder) to 89.2% (generalized anxiety disorder). Compared with patients who had a single mental disorder, patients with co-occurring disorders reported significantly more disability in social and occupational functioning. After adjustment for other mental disorders and demographic and general health factors, compared with patients with no mental disorder, only patients with major depressive disorder, bipolar disorder, phobias, and substance use disorders had significantly increased disability, as measured by the Sheehan Disability Scale. CONCLUSIONS: Primary care patients with more than one mental disorder are common and highly disabled. Individual mental disorders have distinct patterns of psychiatric comorbidity and disability. PMID- 9396955 TI - Premenstrual syndrome: evidence for symptom stability across cycles. AB - OBJECTIVE: This study assessed cycle to cycle symptom stability in women with premenstrual syndrome (PMS). METHOD: Symptom ratings obtained prospectively over three or more symptomatic cycles from 16 women with PMS were analyzed. Measures of symptom severity and change were used to generate a coefficient of variation and an intraclass correlation coefficient (ICC) for each one of 14 symptoms across all cycles. In addition, symptoms were divided into three clusters, and the stability of the rank order of severity of symptoms within clusters and the correlation between symptom clusters were also calculated. RESULTS: In the 65 cycles studied, mood symptoms were the most prevalent. Mood symptoms--anxiety, irritability, and mood lability--had the lowest coefficients of variation but also the lowest ICCs of all symptoms. Within their respective symptom clusters, both physical and mood symptoms showed remarkable stability across cycles of their rank order of severity, but only the mood symptom cluster was highly correlated with functional impairment. CONCLUSIONS: Three mood symptoms--anxiety, irritability, and mood lability--were the most stable symptoms in this group of women with PMS. Mood and not somatic symptoms accounted for most of the functional impairment in this group of women. It is concluded that PMS is a stable syndrome that may best be viewed as part of the spectrum of recurrent mood disorders. PMID- 9396956 TI - Evaluation of a complex case: the value of a drug washout period. PMID- 9396957 TI - Images in psychiatry. The Payne Whitney Clinic. PMID- 9396958 TI - Mood improvement following daily left prefrontal repetitive transcranial magnetic stimulation in patients with depression: a placebo-controlled crossover trial. AB - OBJECTIVE: Preliminary studies have indicated that daily left prefrontal repetitive transcranial magnetic stimulation might have antidepressant activity. The authors sought to confirm this finding by using a double-blind crossover design. METHOD: Twelve depressed adults received in random order 2 weeks of active treatment (repetitive transcranial magnetic stimulation, 20 Hz at 80% motor threshold) and 2 weeks of sham treatment. RESULTS: Changes from the relevant phase baseline in scores on the 21-item Hamilton depression scale showed that repetitive transcranial magnetic stimulation significantly improved mood over sham treatment. During the active-treatment phase, Hamilton depression scale scores decreased 5 points, while during sham treatment the scores increased or worsened by 3 points. No adverse effects were noted. CONCLUSIONS: These placebo controlled results suggest that daily left prefrontal repetitive transcranial magnetic stimulation has antidepressant activity when administered at these parameters. Further controlled studies are indicated to explore optimal stimulation characteristics and location, potential clinical applications, and possible mechanisms of action. PMID- 9396959 TI - Sexual functioning in chronically depressed patients treated with SSRI antidepressants: a pilot study. AB - OBJECTIVE: This prospective study assessed changes in depression and sexual functioning in chronically depressed men and women during treatment with selective serotonin reuptake inhibitors (SSRIs). METHOD: Twenty-five subjects (14 women, 11 men) with DSM-III-R dysthymia, chronic major depression, or double depression were administered the Arizona Sexual Experience Scale and the Hamilton Depression Rating Scale before and after 6 weeks of treatment with sertraline or paroxetine. RESULTS: As measured by scores on the Arizona Sexual Experience Scale, desire, psychological arousal, and overall sexual functioning significantly improved in women; orgasm delay, orgasm satisfaction, and overall sexual functioning significantly worsened in men. CONCLUSIONS: This study suggests that after SSRI treatment, difficulties with desire and psychological arousal in depressed women tend to remit, whereas in men orgasmic dysfunction appears to be a side effect to medication. PMID- 9396960 TI - Emergence of adverse events following discontinuation of treatment with extended release venlafaxine. AB - OBJECTIVE: The rate of adverse events following discontinuation of treatment with extended-release venlafaxine was compared with the rate associated with discontinuation of placebo administration. METHOD: The subjects were 20 outpatients with major depressive disorder who had participated in a multicenter, double-blind, placebo-controlled study of the efficacy of the new extended release formulation of venlafaxine. RESULTS: During the 3 days after discontinuation of treatment with the study drug, seven (78%) of the nine venlafaxine-treated subjects and two (22%) of the nine placebo-treated patients reported the emergence of adverse events, a statistically significant difference. CONCLUSIONS: These results suggest that clinicians discontinuing venlafaxine treatment should consider tapering the medication dose gradually. PMID- 9396961 TI - Plasma levels of cytokines and soluble cytokine receptors during treatment with haloperidol. AB - OBJECTIVE: Clozapine increases the levels of cytokines and soluble cytokine receptors. The authors investigated whether haloperidol has similar effects. METHOD: Rectal temperature, white blood cell counts, and plasma levels of cytokines and soluble cytokine receptors were assessed before and during 6 weeks of haloperidol treatment in 10 psychiatric patients. RESULTS: Haloperidol at mean doses of 7.0 mg/day (SD = 3.4), 6.9 mg/day (SD = 3.4), and 5.0 mg/day (SD = 3.1) at the end of the 1st, 2nd, and 6th weeks of treatment, respectively, did not affect rectal temperature, white blood cell counts, or plasma level of interleukin-1 receptor antagonist, interleukin-6, tumor necrosis factor-alpha (TNF-alpha), soluble TNF receptor p55 or p75, or soluble interleukin-2 receptor. CONCLUSIONS: Haloperidol is unlikely to confound the results of studies investigating disease-related alterations in the levels of a broad range of cytokines and soluble cytokine receptors in schizophrenia. PMID- 9396962 TI - Self-reported anxiety, general medical conditions, and disability bed days. AB - OBJECTIVE: This study examined the effect of self-reported anxiety on the number of days persons with various general medical conditions spend in bed owing to disability. METHOD: Self-reported medical illness and disability data from a nationally representative household survey sample (N = 20,884) were analyzed. RESULTS: Among respondents with general medical conditions, those with self reported anxiety had a nearly fourfold greater length of disability (mean = 18.0 bed days) than the nonanxious respondents (mean = 4.8 bed days). After adjustment for differences in demographic characteristics and burden of general medical illness, anxiety was associated with an additional 3.8 bed days. CONCLUSIONS: Self-reported anxiety in combination with general medical conditions may be associated with extensive functional impairment. PMID- 9396963 TI - Dispositional predictors of problem gambling in male adolescents. AB - OBJECTIVE: This study investigated the possible relationship between impulsivity in early adolescence and gambler status in late adolescence. METHOD: Impulsivity measures consisting of self-reports and teacher ratings were gathered from 754 boys in early adolescence, and their gambling status in late adolescence was assessed with a self-report measure. RESULTS: On both measures of impulsivity, nongamblers had the lowest scores, recreational gamblers had the next higher scores, low problem gamblers had still higher scores, and high problem gamblers had the highest scores. CONCLUSIONS: These findings support the DSM-IV classification of problem gambling as a deficit in impulse control. PMID- 9396964 TI - CSF 5-HIAA and family history of antisocial personality disorder in newborns. AB - OBJECTIVE: The relationship between genetic liability for antisocial behavior and CSF 5-hydroxyindoleacetic acid (5-HIAA) in newborns was explored. METHOD: The authors assayed 5-HIAA in "leftover" CSF from 193 neurologically normal newborns and obtained family psychiatric histories of the newborns' first- and second degree relatives. RESULTS: Levels of 5-HIAA were significantly lower in the infants with family histories of antisocial personality disorder than in the newborns without such family histories. CONCLUSIONS: These findings support the possibility that serotonin mediates one component of genetic liability to antisocial outcome, but the magnitude of that component may be less than what has been inferred from previously published reports. PMID- 9396965 TI - Pindolol-paroxetine combination. PMID- 9396966 TI - Bulimia outcome. PMID- 9396967 TI - Schizophrenia practice guideline. PMID- 9396968 TI - Schizophrenia practice guideline. PMID- 9396969 TI - Genetic and immunologic aspects of cystic fibrosis. AB - LEARNING OBJECTIVES: Reading this article will enable the readers to reinforce their knowledge of the pathophysiology of cystic fibrosis (CF), the pathogenesis of the lung disease, the criteria for diagnosis, and CF genotype/phenotype relationships. The focus of this review is on the genetic and immunologic aspects of CF. DATA SOURCE: Relevant articles, current texts, data presented at the annual North American Cystic Fibrosis Conferences and distributed to the Directors of CF Centers by the CF Foundation were reviewed. A MEDLINE database using subject keywords was searched from 1987 to date. Background information derived from the author's 33 years of clinical experience at three of the CF Foundation's CF Care, Teaching and Resource Centers was also included. STUDY SELECTION: Since CF is an inherited disorder, the genetic aspects are emphasized. With the cloning of the CF gene, DNA analysis has assumed an important role in confirming the clinical diagnosis and in the improved understanding of the pathophysiology of this disorder. Although DNA testing is highly specific, it is not very sensitive. RESULTS: Cystic fibrosis gene structure and function are described briefly. The pathophysiology of CF, as it relates to the CF gene defect, and the current knowledge of the pathogenesis of the lung disease are reviewed. The criteria for the diagnosis proposed by the Clinical Practice Guidelines for CF are discussed. Problems of establishing the diagnosis and the importance of correlations of laboratory and clinical findings in CF are emphasized. CONCLUSIONS: As a multisystem disorder, CF can masquerade as other disorders, including allergic respiratory disease. Primary care physicians often refer patients to allergists/immunologists because of recurrent respiratory problems. This review discusses the genetic heterogeneity of CF. PMID- 9396970 TI - Recurrent facial angioedema with elevated antinuclear antibodies. PMID- 9396971 TI - IgE-binding components of staphylococcal enterotoxins in patients with atopic dermatitis. AB - BACKGROUND: The exacerbation of atopic dermatitis may be associated with infection of the skin with Staphylococcus aureus (S.aureus). S. aureus isolated from the skin of patients with atopic dermatitis secretes enterotoxin A, B, and toxic shock syndrome toxin 1. This is of interest because these patients may develop specific IgE antibodies against components from staphylococci. OBJECTIVE: The objective was to demonstrate IgE-sensitization to components of Staphylococcus aureus enterotoxins A and B (purified and partially purified), toxic shock syndrome toxin 1, and the bacterial cell component lipoteichoic acid, in patients with atopic dermatitis. METHODS: Blood samples from 34 patients with atopic dermatitis and 10 controls were tested by leukocyte histamine release to the enterotoxins and lipoteichoic acid. The toxins were separated by sodium dodecylsulfate polyacrylamide gel electrophoresis and analyzed by IgE immunoblotting with sera from the same patients. RESULTS: The majority of patients (96%) with clinical signs of skin infection produced specific IgE antibodies to all three toxins. Nearly half of the patients produced IgE to enterotoxin A and B. Only 63% of the patients with atopic dermatitis showed cellular response judged by the release of histamine from patient basophils when challenged in vitro with the toxins. This may indicate clinically unimportant sensitization in a number of patients. The immunoblotting revealed that the major allergens of the toxins were 24 and 28 kD proteins. Partially purified toxins showed a higher frequency of leukocyte histamine release responses than purified toxin. The only obvious difference was a difference in the content of pure toxin of the two preparations. Lipoteichoic acid showed nonspecific activity. CONCLUSION: These findings suggest that staphylococcal enterotoxins may act as specific allergens and induce IgE-antibodies to enterotoxins that may exacerbate the skin inflammation in some patients with atopic dermatitis. PMID- 9396972 TI - Trimethoprim/sulfamethoxazole incremental dose regimen in human immunodeficiency virus-infected persons. AB - BACKGROUND: The mechanism of tolerance to incremental doses of trimethoprim sulfamethoxazole given to human immunodeficiency virus-infected persons who have had a prior intolerance to this agent has not been studied. OBJECTIVE: We prospectively evaluated a regimen of incremental doses of oral trimethoprim sulfamethoxazole in human immunodeficiency virus-infected persons who had a prior trimethoprim-sulfamethoxazole-induced fever and nonexfoliative skin rash to investigate the mechanism by which it permits tolerance. METHODS: Oral trimethoprim (0.00004 mg)/sulfamethoxazole (0.00002 mg) was given to 22 human immunodeficiency virus-infected persons on day 1 and gradually increased over eight days to 1 double strength (DS) tablet/day in an outpatient setting. At study entry, skin tests and IgG antibodies to sulfa were performed; the latter was repeated at study week 4. RESULTS: Nineteen patients tolerated trimethoprim/sulfamethoxazole at the completion of the 8-day protocol (86% effective). Moderate toxicities occurred in eight persons during the desensitization protocol; five of these were able to continue trimethoprim/sulfamethoxazole with adjunctive prednisone. Skin tests to sulfa antigen were negative in all persons. Eleven patients at study entry had antibodies to sulfamethoxazole; IgG antibodies appeared at week 4 in 8 of the 11 patients who initially had no antibody detected. CONCLUSIONS: The mechanism of tolerance to the incremental doses of trimethoprim/sulfamethoxazole given to previously intolerant human immunodeficiency virus-infected persons is not due to desensitization and remains undetermined. PMID- 9396973 TI - Popsicle-induced anaphylaxis due to carmine dye allergy. AB - BACKGROUND: IgE-mediated hypersensitivity is a suggested mechanism to explain adverse reactions from carmine-containing products. OBJECTIVE: To describe a patient who experienced anaphylaxis after ingestion of a popsicle colored with carmine and to provide additional evidence that the adverse reaction was IgE mediated. METHODS: The patient and her husband underwent skin prick tests to the popsicle and carmine. The patient also received skin prick tests and/or open oral challenge to each of the other components of the incriminated food. Topical application of cosmetics with and without carmine to the patient's forearm was also performed. To confirm carmine-specific IgE, a Prausnitz-Kustner (P-K) test was performed using the patient's husband as recipient. Twenty control subjects also were tested to carmine by skin prick test. RESULTS: The patient showed 4+ skin prick test responses to the popsicle and carmine. Skin prick tests and/or open oral challenge to each of the other components of the popsicle were negative. The patient's husband's and 20 control subjects' skin prick tests to carmine were negative as was the patient's husband's skin prick test to the popsicle. Skin prick test reactivity to the popsicle and carmine were successfully transferred to the patient's husband in P-K format. Cosmetics applied to the patient's forearm elicited no immediate response. CONCLUSION: The positive skin prick tests to the popsicle and carmine and the successful (P-K) transfer of skin prick test reactivity support a carmine-specific, IgE-mediated mechanism in explaining our patient's popsicle-induced anaphylaxis. PMID- 9396974 TI - Localized unilateral periorbital edema induced by aspirin. AB - BACKGROUND: Aspirin intolerance manifested as bronchospasm or urticaria/angioedema has been observed since the beginning of this century. OBJECTIVE: To report a novel case of intolerance to aspirin ingestion. METHODS: Case report; routine skin testing; pulmonary function testing; aspirin challenge. RESULTS: A 30-year-old man with a history of left ocular trauma at the age of 10 noted a 3-year history of left periorbital angioedema after aspirin but not other nonsteroidal anti-inflammatory drugs. Incremental oral aspirin challenge resulted in this unilateral symptomatology at a dose of 673 mg. CONCLUSION: To the best of our knowledge, this is the first reported case of unilateral periorbital edema following aspirin ingestion. PMID- 9396975 TI - Asthma mortality in Mauritius: 1982-1991. AB - BACKGROUND: Bronchial asthma is a common problem in the island of Mauritius and its prevalence seems to be increasing. OBJECTIVE: In order to appreciate the magnitude of the problem, patterns of asthma mortality were studied during a period of 10 years. METHOD: All death certificates issued in the island from 1982 to 1991 were reviewed and all cases of asthma deaths were selected. RESULTS: The global asthma mortality rate was found to be 20/100,000 in 1982, and it decreased to 12/100,000 in 1991. Similarly the asthma death rate for the 0 to 4 year age group decreased from 20/100,000 in 1982 to 5/100,000 in 1991. For the 5 to 34 year age group, it decreased from 2.6/100,000 in 1982 to 1.02/100,000 in 1991. There was no statistically significant difference between the various ethnic groups. CONCLUSION: Our study showed that in a developing country such as Mauritius asthma death rates may be high but may show decreasing trends. Nevertheless, it is generally perceived that the prevalence of the disease is increasing. PMID- 9396976 TI - Skin prick reaction and nasal provocation response in diagnosis of nasal allergy to the house dust mite. AB - BACKGROUND: The allergen skin test is commonly used to ensure the diagnosis of allergic rhinitis even though positive results do not necessarily indicate that rhinitis is of allergic origin. OBJECTIVE: To determine the association between skin prick reactions and nasal provocation responses to Dermatophagoides pteronyssinus (Der p) allergen extract. METHODS: Twenty-six patients with perennial rhinitis and 25 controls underwent skin prick and nasal provocation tests to standardized Der p allergen extract. With the use of allergen extract titration delivered by a metered dose pump, nasal stuffiness, itching, and sneezing were noted, the amount of secretions measured, and nasal airway resistance was recorded by active anterior rhinomanometry. RESULTS: The majority of the patients with rhinitis (20/26), but none of the controls, exhibited strong skin test positivity (4+) to Der p allergen extract. In addition, the majority of the patients with 4+ skin reactions (16/20) had moderate to severe rhinitis. Significantly increased nasal reactivity to the allergen was also observed among those with 4+ skin test positivity. The controls exhibited nasal provocation responses only with significantly higher end-point doses of the allergen extract regardless of the skin test results. CONCLUSION: Only 4+ skin test positivity was closely associated with increased nasal reactivity to Der p allergen among the patients with perennial rhinitis. The nasal provocation technique would be a useful adjunct testing to ensure the diagnosis of nasal allergy to the Der p mite, particularly among those patients with rhinitis with only mild to moderate skin test positivity. PMID- 9396977 TI - Allergenic fungi in allergic fungal sinusitis. AB - BACKGROUND: Patients with allergic fungal sinusitis demonstrate skin test reactivity to many fungal extracts. Various fungi have been isolated from the characteristic allergic mucin. OBJECTIVE: This study was designed to identify allergens in allergic mucin and to compare them to those found in commercial fungal extracts. METHODS: Allergic mucin was collected during functional endoscopic sinus surgery from 11 patients meeting strict diagnostic criteria for allergic fungal sinusitis and from three allergic rhinitis patients with chronic sinusitis. A portion was solubilized in saline and centrifuged. To identify allergens, proteins in allergic mucin and fungal extracts were separated by SDS polyacrylamide gel electrophoresis, transferred to nitrocellulose and immunostained using patient sera and enzyme-labeled anti-human IgE. RESULTS: All patient sera recognized numerous bands ranging from 18 to 90 kD. In mucin, bands were consistently found in the 35 to 50 kD range. Corresponding bands in fungal extracts were found in only 1/11 patients with allergic fungal sinusitis. Sera from 4/11 patients detected an 18-kD protein in allergic mucin, but sera from all patients with allergic fungal sinusitis recognized an 18-kD protein in commercial fungal extracts. Sera from selected patients with allergic fungal sinusitis detected human epithelial proteins in the 35 to 50 kD range. CONCLUSIONS: Fungal allergens were not detected in allergic mucin of all patients with allergic fungal sinusitis. The 18-kD allergen appears to be shared by many fungi, and may be a fungal panallergen. The source of the apparent allergens in the 3550-kD range warrants further study. PMID- 9396978 TI - Dust mite allergen avoidance in the treatment of hospitalized children with asthma. AB - BACKGROUND: Asthma is a leading cause of hospital admission in children. The majority of children with asthma are sensitized and exposed to inhalant allergens that may contribute to chronic airway inflammation. OBJECTIVE: To evaluate the practicality and effects of dust mite (D. farinae and D. pteronyssinus) allergen avoidance in homes of children hospitalized with acute asthma. METHODS: Children 5 to 18 years of age who were admitted with asthma to a suburban Atlanta hospital were randomly assigned, without knowledge of allergen sensitization or exposure in their houses, to active (n = 13) or placebo (n = 10) treatment group. Active treatment included encasing mattress, box springs, and pillows in allergen impermeable covers; weekly hot water wash of bed linens; replacement of bedroom carpet with polished flooring; and 3% tannic acid spray to living room carpet. Placebo treatment included permeable encasing for bedding, cold water wash, and water spray for carpet. Dust samples were analyzed for dust mite, cockroach, and cat allergens, while serum samples were analyzed for IgE antibodies to the same allergens. Outcome measures included daily peak expiratory flow rates, spirometry, methacholine inhalation challenge, and hospital readmission. RESULTS: Children in both groups were similar by demographics, sensitization, and exposure to dust mite allergen. Allergen levels fell > 3-fold in many active and placebo homes. Children in the active group had improved PEFR at 3 and 6 months after intervention (P < .04, P < .05, respectively). Six of seven children in the study who were sensitized and exposed to dust mite allergen demonstrated improved PEFR at 3 months when allergen levels fell in both bedding and bedroom floor. There was no difference in FEV1 or methacholine challenge, although a few children in either group could tolerate methacholine because of bronchial hyperreactivity. Six children (four active and two placebo) were readmitted to hospital during the study. CONCLUSION: Increases in PEFR were recorded among children in the active treatment group and also among sensitized patients whose dust mite allergens fell. These results support the hypothesis that avoidance can be effective even among children admitted to hospital. The study was complicated by insufficient numbers of mite-allergic children and poor compliance with diaries and the protocol. Recruitment from the hospital resulted in participants with more severe asthma than anticipated. The results also suggest that many of the patients in this group will continue to have exacerbations triggered by upper or lower respiratory tract infections. PMID- 9396979 TI - Efficacy and safety of fexofenadine hydrochloride for treatment of seasonal allergic rhinitis. AB - BACKGROUND: H1-receptor antagonists are effective for the treatment of seasonal allergic rhinitis. In rare circumstances, some second-generation H1-receptor antagonists have been associated with prolongation of the corrected QT interval (QTc), thus increasing the risk of ventricular arrhythmias. Fexofendine HCl, the carboxylic acid metabolite of terfenadine, is a new second-generation antihistamine that is nonsedating and does not cause electrocardiographic effects. OBJECTIVE: To investigate the clinical efficacy and safety of fexofenadine HCl in the treatment of ragweed seasonal allergic rhinitis and to characterize the dose-response relationship of fexofenadine HCl at dosages of 60, 120, and 240 mg bid. METHODS: A multicenter, 14-day, placebo-controlled, double blind trial was conducted with patients suffering from moderate to severe ragweed seasonal allergic rhinitis who met symptom severity criteria after a 3-day placebo baseline period. Patients with minimal or very severe symptoms during the baseline period were excluded. Patients were randomized to receive fexofenadine HCl (60, 120, or 240 mg bid) or placebo at 12-hour dosing intervals (7:00 AM and 7:00 PM). The primary efficacy measure was patient-assessed 12-hour reflective total symptom score before the evening dose (trough). RESULTS: Five hundred seventy patients completed the trial. Fexofenadine HCl at each dosage provided significant improvement in total symptom score (P < or = .003) and in all individual nasal symptoms compared with placebo. The frequency of adverse events was similar among fexofenadine HCl and placebo groups, with no dose-related trends. No sedative effects or electrocardiographic abnormalities, including prolongations in QTc were detected. CONCLUSIONS: Fexofenadine HCl is both effective and safe for the treatment of ragweed seasonal allergic rhinitis. Because there was no additional efficacy at higher dosages, 60 mg bid appears to be the optimal therapeutic dosage for these patients. PMID- 9396980 TI - Occupational respiratory allergic disease induced by Passiflora alata and Rhamnus purshiana. AB - BACKGROUND: There has been an increase in the incidence of occupational asthma along with better understanding of its pathophysiologic mechanisms and etiologic factors. There are no reports of patients with asthma and rhinitis to Passiflora alata (passion flower) and Rhamnus purshiana (cascara sagrada). METHODS: We describe two substances of plant origin as causal agents of occupational allergic respiratory diseases in a patient who worked in a pharmacy devoted to the manual preparation of products. RESULTS: Skin testing and Western blot confirmed the sensitization of the patient to these plant extracts in vivo and in vitro, respectively. Bronchial challenge confirmed the cause-effect relationship between the allergen exposure and the diseases. CONCLUSIONS: We conclude that Passiflora and cascara sagrada are two new etiologic agents of IgE-mediated occupational asthma and rhinitis. The present study also serves to alert physicians to the risks associated with work in pharmacies devoted to manual preparation of plant extracts, emphasizing the importance of the use of protective measures in these environments. PMID- 9396981 TI - Sensitization to indoor allergens and the risk for asthma hospitalization in children. AB - BACKGROUND: Hospitalization for asthma continues to present a major health problem despite advances in our understanding that asthma is an inflammatory disease of the bronchi and that exposure to specific allergens can induce and worsen this inflammation. The role of sensitization to specific indoor allergens and hospitalization for acute asthma in children is unclear. OBJECTIVE: The purpose of this study was to evaluate the independent contributions of sensitization to specific indoor allergens among children with asthma to the risk of hospitalization for asthma. METHODS: The charts of 138 consecutive children with asthma, aged 5 to 18 years, seen at pediatric allergy clinics were reviewed to obtain the results of skin tests to cat, dog, cockroach, and dust mite allergens and the history of hospitalization for asthma within the year prior to the clinic visit. Logistic regression analysis was used to examine the association between indoor allergen sensitivity and other factors, and the risk of hospitalization for asthma. RESULTS: In univariate analyses, hospitalization for asthma was significantly associated with cockroach sensitivity (odds ratio [OR] = 2.2; 95% confidence interval [CI] = 1.1, 4.3); cat sensitivity (OR = 2.9; CI = 1.3, 6.4); black race (OR = 2.4; CI = 1.1, 5.1); public aid/self pay (OR = 2.3; CI 1.1, 4.9) and age, (OR [per year increase in age] = 0.8; CI = 0.7, 0.9). In a stepwise multiple logistic regression analysis, only cat sensitivity (OR = 3.8; CI = 1.5, 9.2), age (OR = 0.8; CI = 0.7, 0.9) and race (OR = 3.2; CI = 1.4, 7.5) entered into the model as significant independent predictors. CONCLUSION: Sensitivity to cat allergen may be an important determinant for asthma hospitalization in children. Sensitization to cockroach allergen per se was not found to be an independent risk factor. As observed in previous studies, younger and black children were at increased risk of hospitalization for asthma. PMID- 9396982 TI - Follow-up of pediatric patients with recurrent infection and mild serologic immune abnormalities. AB - BACKGROUND: Children with recurrent infections significant enough to warrant referral to an immunologist frequently have mild abnormalities of the humoral immune system. Parents of these children are generally reassured that their child will outgrow the clinical problems that prompted their referral. OBJECTIVE: This is a retrospective study with the objective being to evaluate changes in immune measurements and clinical status of children with recurrent infections followed in an immunology clinic. METHODS: Forty-two patients (mean age 60 months) previously evaluated for recurrent infections were re-evaluated after at least 12 months (mean 37 months). Initial evaluation studies included quantitative immunoglobulins in all patients and IgG subclass determinations and pre- and postvaccination pneumococcal polysaccharide antibody titers in a subpopulation of patients. RESULTS: Patients were assigned to one of two categories: those with initial laboratory abnormalities (27 patients) and those with normal initial studies (15 patients). Among the patients with initial abnormalities, partial IgA deficiency was most commonly seen (20/27). It persisted in 15. Only 6/27 patients had studies that were completely normal on follow-up. Among patients with no initial abnormality, 9/15 developed a partial deficiency of at least one immunoglobulin isotype or IgG subclass. Eighty-six percent of the patients were clinically improved at the time of their last follow-up visit regardless of their laboratory values. CONCLUSIONS: A high proportion of children with recurrent infection have persistent, partial immunoglobulin deficiencies lasting in some cases for years. Despite this finding almost all patients demonstrate clinical improvement with time. PMID- 9396983 TI - Comparison of ebastine with cetirizine. PMID- 9396984 TI - Hepatitis C after intravenous immunoglobulin. PMID- 9396985 TI - Lack of ragweed in northwestern US. PMID- 9396986 TI - Clarification of Richard C. Ahren's editorial commenting on article by Pallares DE et al. PMID- 9396987 TI - Quality of life after surgery for early breast cancer. PMID- 9396988 TI - Quality of life after breast conservation or mastectomy: a systematic review. AB - BACKGROUND: For early breast cancer, survival after breast conservation is similar to that after mastectomy. Some women may not have a clear preference and wish to have further information about quality of life experienced after the alternative treatments. This paper describes a systematic review of randomized trials on mastectomy versus breast conservation for which there are data on quality of life or psychosocial outcomes. METHODS: Literature was reviewed to find all randomized controlled trials comparing breast conservation to mastectomy, with quality of life or psychological effects as an outcome. Studies were then critically appraised by two reviewers independently and any disagreements about their quality and results resolved by discussion. RESULTS: A total of six randomized trials met our inclusion criteria. In general, they are of poor quality. Women who had breast conservation had a more favourable body image of themselves than those who had mastectomy in all five studies in which it was examined. The evidence was statistically inconclusive for all the other dimensions measured, namely perceptions of psychological health, sexual health, physical health, fear of the future and global quality of life. Radiotherapy may be a determinant of poorer psychological health and body image. CONCLUSIONS: Apart from body image, it is unclear whether breast conservation or mastectomy results in better psychosocial outcomes. Moreover, the studies were done before evidence was available to inform women about the equivalence of survival with these alternative treatments. Therefore there is inadequate information available to help many women decide about their choice of treatment in the future. Preference trials should be conducted, using standardized quality-of-life measures, in which women who are uncertain about which treatment to choose are randomized to breast conservation or mastectomy. PMID- 9396989 TI - Breast cancer: aetiological factors and associations (a possible protective role of phytoestrogens). AB - BACKGROUND: In spite of many known and suspected factors associated with the risk of breast cancer there has until recently been no explanation for its continuing increase in women of Western societies over recent decades or why there has not been an equivalent increase in women of most Asian and other less Westernized societies. It has long been suspected that a significant factor has been an increasing change of diet in Western societies from one predominantly vegetarian to one with a high content of meat and dairy products as well as 'refined' foods. Although diet has long been suspected there has otherwise been no real explanation as to the mechanism of the change in incidence of breast cancer. METHODS: A comprehensive literature review has been made of aetiological factors and associations concerning breast cancer to determine whether any consistent trend can explain the rising incidence in Western societies. RESULTS: There are a number of likely contributory factors but there is now accumulating evidence that the single most important difference is that people having a vegetarian diet have a high intake of legumes and other plant foods containing a variety of lignans and isoflavonoids. These appear to have an important role as nature's sex hormone modulators. These agents appear to be biologically active in a number of ways not yet completely understood but they do have both a weak oestrogenic effect and an anti-oestrogenic competitive effect, thus reducing the potential carcinogenic action of prolonged oestrogen activity. A probable additional benefit of such diets could be the role of dietary fibre. CONCLUSIONS: A major problem of Western diets may not be the presence of meat or dairy products in the diet but the absence of desirable ingredients of vegetarian diets, namely dietary fibre and certain plant lignans and isoflavonoids. A modification of diet to include a greater proportion of fibre and soy or other leguminous plant food should be studied. Alternatively addition of more fibre and lignans and especially isoflavonoids to traditional Western diets would seem worthy of serious investigation. Such influences appear to have their greatest impact early in life and therefore could be especially important for girls and young women in Western societies. PMID- 9396990 TI - Breast cancer in young women. AB - BACKGROUND: It is believed that cancer of the breast is more difficult to diagnose in young women and it has long been disputed whether breast cancer occurring in women aged < or = 40 years is more aggressive than that occurring later in life. A number of reports in the literature suggest that the disease is of similar aggressiveness in the young patients and older age groups, while other reports suggest that it is more aggressive and carries a higher mortality in young women. METHODS: To address these aspects of breast cancer we have undertaken a review of the cases treated at The Strathfield Breast Centre between 1989 and 1996 and compared the disease in the young and old groups with particular reference to the modes of diagnosis, the pathological staging and types of tumour and the outcomes of treatment. RESULTS: The accuracy of ultrasound and fine needle aspiration biopsy were similar in both groups, but the false negative rate of mammography in the young patients was 15% or 50% greater than that which was observed in the older patients. The incidence of histopathological type, bilaterality, size of lesion and receptor positivity were the same in both groups. In the young group, 40% had Grade 3 tumours compared with 27% in the older group. Nineteen per cent of young patients had 4 or more lymph nodes involved while only 10% of the older patients had similar lymph node involvement. Overall 5-year survival was 79% in the older patients compared with 90% in the young patients. CONCLUSIONS: The spectrum of disease is similar in both the young and older patient and the prognosis is no worse for the young group but mammography is less effective in the diagnosis of the young patient. PMID- 9396991 TI - A simple index to predict prognosis independent of axillary node information in breast cancer. AB - BACKGROUND: Since the course of breast cancer is often unpredictable, we wished to develop a model using characteristics of the primary tumour alone to predict prognosis. METHODS: Several tumour features were determined, and after a median follow-up duration of 65 months, multivariate analysis identified tumour size and grade, oestrogen receptor concentration, axillary lymph node metastasis and tumour cell proliferation fraction (MIB-1 count) as being independently associated with increases in risk for both relapse and death from breast cancer. A prognostic model was constructed using tumour size and grade, oestrogen receptor concentration and MIB-1 count only. A score of 1 for each was given to tumour size > 20 mm, tumour grade 2 or 3, oestrogen receptor concentration < 10 fmol/mg cytosol protein and MIB-1 count > 9%. Five groups established by assigning a combined score of 0, 1, 2, 3 or 4 for each patient were analysed for their associations with disease-free and overall survivals. RESULTS: This preliminary model predicted 5-year survival rates of 97, 91, 85, 68 and 50% for the five groups. The model was further simplified by excluding tumour grade from the analysis. The revised model identified four risk groups with predicted 5-year survival rates of 91, 86, 66 and 52%. This model, the Adelaide prognostic index, was also able to identify four risk groups in both node-negative and node positive patients. CONCLUSIONS: The Adelaide prognostic index can be used to predict prognosis even in the absence of axillary lymph node information. PMID- 9396993 TI - Communicating with patients: surgeons' perceptions of their skills and need for training. AB - BACKGROUND: This study assessed surgeons' current perceived level of competence in a number of interactional skills, their perceptions of the need for training and assessment in interactional skills, and their perceptions of the appropriateness of the format and content of two existing communication skills training packages. METHODS: Of 267 surgeons who were sent the survey, 63% (n = 143) of eligible respondents completed and returned it. RESULTS: More than three quarters of the sample identified the following skills as being important or very important in being a good surgeon: breaking bad news; preparing patients for surgical procedures; educating patients about their diagnosis and treatment, and increasing the likelihood that they will remember what they have been told; detecting anxiety and depression in patients, encouraging patients to express these and listening to their anxieties. More than half the sample felt at least competent at seven of the 10 interactional skills, but almost one-third of the sample reported being 'not or not at all competent' at increasing patients' ability to remember what they have been told and at encouraging patients to express anxieties about their condition, and a further 13.3% reported a lack of competence at breaking bad news to patients about their diagnosis/prognosis. A higher proportion reported a lack of competence in providing bereavement counselling (59.6%), and gaining consent for organ donation (56.6%) and for autopsy (48.9%). The majority rated different aspects of the two communication skills training packages as either 'good' or 'excellent'. CONCLUSIONS: The survey identified a number of communication skills which are perceived by surgeons to be important and to require formal training and assessment. PMID- 9396992 TI - Cosmetic outcome of breast-conserving therapy in Chinese patients with early breast cancer. AB - BACKGROUND: Breast-conserving surgery combined with radiotherapy has emerged as an alternative to mastectomy for women with early breast cancer, and cosmetic outcome has correlated closely with the psychosocial and physical well-being of the patient. Cosmetic outcome assessment after breast-conserving therapy in Chinese patients has so far not been conducted among the clinicians, the patients or their spouses. METHODS: The cosmetic results from breast-conserving therapy were evaluated in a group of 33 patients who had been selected as suitable for undergoing local excision, axillary dissection and postoperative radiation therapy for early stage breast cancer. The success of the procedures was assessed by the patients, the clinicians and the patient's spouse, and their ratings were compared with each other. RESULTS: Eighty per cent of the patients and their spouse were satisfied with the cosmetic outcome. Using McNemar's test, when the groups were evaluated on a case-by-case basis, there was a good level of concordance between the patients and their spouses, and that of the patients and the clinicians. CONCLUSIONS: Evaluation of the cosmetic and psychosocial sequelae of breast cancer patients is essential when new approaches to treatment are introduced; our data suggest that cosmetically successful breast conservation is feasible in a selected group of Chinese women with early breast cancer. PMID- 9396994 TI - Prostate cancer mortality in New Zealand: the past and projections for the future. AB - BACKGROUND: Ageing male populations and improved diagnosis of early stage disease have contributed to the increasing incidence of prostate cancer observed in many Western countries. The clinical significance of these diagnosed cancers is, however, currently unclear. The aim of this study is to examine trends over time in prostate cancer mortality as an indicative measure of clinically significant disease during a 29-year period (1965-93) which preceded the extensive use of early cancer diagnostic techniques or radical therapy protocols. METHODS: Age specific and age-standardized rates were calculated for each year during the study period, using routinely collected mortality and demographic data. A Poisson regression model was used to describe trends in the age-specific rates over time to predict numbers of prostate cancer deaths and the lifetime risk of death over the next 20 years. RESULTS: Significant annual increases ranging from 1 to 2.6% were found for age-specific prostate cancer mortality rates over the 29-year time period, with the largest increases experienced in the younger age groups at risk. Based on projected population ageing and growth alone, annual numbers of prostate cancer deaths are predicted to increase from 487 in 1993 to 664 by the year 2006 and then to 833 by the year 2016. Continuation of the observed increases in age specific mortality rates would result in a predicted 797 deaths by the year 2006, while an expected 1115 deaths is calculated for the year 2016. This would correspond with an increase in the lifetime risk of death from prostate cancer from a present 3.7 to 4.5% in 10 years and 6.2% in 20 years. CONCLUSIONS: The changing pattern of prostate cancer mortality described in this study is likely to represent a significant increase in the incidence of clinically significant disease. This will have a significant impact on the ageing New Zealand male population, and important implications for the provision of effective treatment and preventive strategies. PMID- 9396995 TI - Fine needle aspiration biopsy in children. AB - BACKGROUND: Fine needle biopsy (FNB) in children has been slow to gain acceptance compared with the use of the technique in adults where it is regarded as standard clinical practice in screening significant lymphadenopathy and suspicious masses. We report our early experience with FNB in the paediatric population. METHODS: Fifty-two biopsies were performed between June 1991 and June 1993. The age of the children ranged from 6 months to 14 years (median 2 years, mean 5 years). RESULTS: A definite diagnosis on cytology alone was obtained in 67%. The pathologist was certain of malignant or nonmalignant potential in 79% (67% benign and 12% malignant) and unsure in 21% (17% benign and 4% malignant). There were no false positive or false negative diagnoses of malignancy. Surgical excision or biopsy was performed in 33%. Fine needle biopsy assisted in planning surgery in 12%. Surgery was necessary for a definite diagnosis in 21% and FNB assisted 42% of the patients to avoid surgery altogether. CONCLUSIONS: Fine needle biopsy is simple, minimally invasive and useful in the evaluation of children with suspicious lymph nodes and masses. PMID- 9396996 TI - Diagnostic laparoscopy for chronic right iliac fossa pain: a pilot study. AB - BACKGROUND: The aim of this study was to determine the value of diagnostic laparoscopy in patients with chronic right iliac fossa pain. METHODS: A retrospective study at Echuca Hospital involving case-note review and telephone questionnaire of patients who had undergone diagnostic laparoscopy for chronic right iliac fossa pain at least 12 months earlier (September 1992 to August 1995) was carried out. RESULTS: Forty-one cases were identified and followed up 12-40 months postoperatively (median 21 months). Eleven cases had positive findings at laparoscopy, of whom eight obtained lasting relief after treatment. Of the remaining 30 patients 17 had a normal-looking appendix removed and 12 were cured; these were younger patients with episodic symptoms and localized signs. Of eight patients who had adhesions divided, four with adhesions beneath old scars obtained relief. Altogether 32 of the 41 patients considered the laparoscopy worthwhile even though in some cases it did not relieve their chronic pain. CONCLUSIONS: Diagnostic laparoscopy is worthwhile for patients with chronic right iliac fossa pain. Concurrent appendicectomy should be considered in young patients with episodic, well-localized symptoms associated with systemic malaise while adhesiolysis may be beneficial for viscero-parietal adhesions beneath abdominal wall scars. PMID- 9396997 TI - Predicting poor outcome in perforated peptic ulcer disease. AB - BACKGROUND: Despite modern medications for peptic ulcers, patients frequently require emergency surgery for complications of ulcer disease. Many of these patients have coexisting medical problems which not only predispose to perforated ulcer disease, but also influence the clinical outcome. This study reviews the outcome of a group of patients with perforated ulcer disease and examines the influence of a range of comorbidity factors on the outcome. METHODS: A retrospective chart review of all cases of perforated peptic occurring over a period of 9 years. RESULTS: One hundred and forty-nine perforated peptic ulcers in 147 patients were diagnosed between 1987 and 1996. Coexisting malignancy, use of immunosuppressives or corticosteroids, pre-operative shock and admission to intensive care were all significantly associated with reperforation by univariate analysis. However, logistic regression analysis indicated that none of these factors independently predicted reperforation which, therefore, occurs as a multifactorial event with all the above factors contributing. Death from perforated ulcer disease was related to pre-operative shock, malignancy, admission to intensive care and reperforation when examined by univariate analysis. Furthermore, logistic regression analysis showed that coexisting malignancy and reperforation were significant predictors of mortality. CONCLUSIONS: Perforated peptic ulcer disease remains a frequent clinical problem in patients with short dyspeptic histories, who may or may not have been using ulcerogenic medications. It is a significant cause of morbidity and mortality among an often aged and otherwise unwell group of patients. Patients with underlying malignant disease, who may be immunosuppressed with corticosteroids or cytotoxics, are at increased risk of dying from perforated ulcer disease. Reperforation of an ulcer, following simple closure or conservative treatment, is also highly predictive of increased mortality. PMID- 9396998 TI - Previous intravenous chemotherapy does not alter response rate or survival time of patients with hepatic metastases from colorectal cancer treated by hepatic artery chemotherapy. AB - BACKGROUND: The present paper addressed the issue of whether pretreatment with intravenous (i.v.) chemotherapy affects response rate or survival in patients receiving hepatic artery chemotherapy (HAC). METHODS: Case note reviews of 164 patients treated in a teaching hospital from June 1990 to July 1996 were carried out. RESULTS: The response rate and carcino-embryonic antigen (CEA) fall in the two groups was almost identical. There was a nonsignificant survival advantage in the non-pretreatment group. CONCLUSIONS: Previous administration of i.v. chemotherapy did not affect the CEA response of patients receiving HAC. PMID- 9396999 TI - Benign anterior knee cyst in early childhood. AB - BACKGROUND: Anterolateral knee cysts are not uncommon in the first 7 years of life, but have not been described in the literature. METHODS: Four patients presenting with an asymptomatic lump on the anterolateral joint line were reviewed. RESULTS: The lump remains asymptomatic. CONCLUSIONS: Anterolateral knee cysts of childhood are benign and do not require treatment. Their cause is conjectural. PMID- 9397000 TI - Laser photocoagulation in the treatment of malignant dysphagia. AB - BACKGROUND: Dysphagia secondary to carcinoma of the oesophagus and gastric cardia is the principal symptom requiring palliation in those patients who present with late-stage disease or who are unfit for surgery. The primary aim of the present study was to determine the safety and efficacy of laser photocoagulation in the palliation of malignant dysphagia. Secondary aims were to look at reasons for failure and predictors of outcome; to determine the most appropriate second line therapy for treatment failures; and to look at the results of treatment for early stage disease. METHODS: Sixty-seven patients treated over a 6-year period with endoscopic Nd:YAG laser photocoagulation were evaluated and the quality of swallowing assessed before and at intervals after treatment. RESULTS: Ninety per cent of patients achieved successful initial palliation. This was sustained in 76% after 3 months of treatment. Within a month before death 71% of patients were palliated but 29% required the addition of second-line treatment to achieve this. Complications were infrequent. There were no deaths attributable to laser treatment. Five of 10 patients treated with radiotherapy developed fibrous stricturing that required endoscopic dilatation. No variables were independently predictive for treatment failure. Six patients with early stage disease experienced prolonged survival. CONCLUSIONS: We conclude that laser photocoagulation offers safe and effective palliation of malignant dysphagia in this group of patients and is appropriate as first-line therapy. PMID- 9397001 TI - The management of colorectal perforation and peritonitis. AB - BACKGROUND: Surgical outcomes in patients presenting with colonic perforation or peritonitis tend to be poor. This study was undertaken to determine outcomes in such patients at a time before multiple re-laparotomies were performed. METHODS: Retrospective analysis of computer records of all patients presenting acutely to the University Surgical Unit (Wellington School of Medicine) with colonic perforation or peritonitis over a 15-year period. RESULTS: Seventy-three patients, 33 males and 40 females were admitted with either perforation or localized peritonitis of colorectal origin. Of these, 78% were managed as emergencies, but six were admitted electively and found incidentally. Consultant surgeons performed surgery slightly more frequently than registrars. Two patients were managed non-operatively. Forty-one per cent received peri-operative blood transfusion and 22% peri-operative total parenteral nutrition. The majority of patients presented with either peritonitis or free perforation in association with diverticular disease. The site of perforation was either ileocolic or sigmoid colonic in the majority of patients. Hartmann's operation was the most commonly performed resection. Respiratory, urinary and wound infections were the most commonly observed postoperative complications. Two patients developed anastomotic leaks (6.3%). The overall persistent intra-abdominal infection rate was 5.5%. Seven patients died following surgery. CONCLUSIONS: Resection of the perforated bowel is mandatory and this should be followed by anastomoses in the case of right-sided lesions and a Hartmann's operation or resection, colostomy and mucous fistula in distally situated lesions. PMID- 9397002 TI - Thirty years experience with heart valve surgery: isolated mitral valve replacement: comment. PMID- 9397003 TI - Thirty years experience with heart valve surgery: isolated aortic valve replacement: comment. PMID- 9397004 TI - Gastrointestinal lavage reduces total body water: comment. PMID- 9397005 TI - Psychiatric disturbance and acute stress responses in surgical patients: comment. PMID- 9397006 TI - The making of a rural surgeon: comment. PMID- 9397007 TI - Ganglioneuroma of the adrenal medulla. PMID- 9397008 TI - Haemorrhage into bronchopulmonary sequestration following cardiac surgery. PMID- 9397009 TI - Localized fibrous tumour of the pleura: two case reviews. PMID- 9397010 TI - MEN type 2a presenting as an intra-abdominal emergency. PMID- 9397011 TI - Perfusion patterns during temporal lobe seizures: relationship to surgical outcome. AB - We sought to determine whether patterns of ictal hyperfusion demonstrated using [99mTC]HMPAO (hexamethylpropylene amine oxime) single photon emission computed tomography (SPECT) predict outcome of temporal lobectomy; in particular, whether the more extensive patterns of ictal hyperperfusion are associated with poor outcome. We studied 63 patients who had ictal SPECT studies prior to temporal lobectomy. Hyperperfusion on ictal SPECT scans was lateralized, and classified into: (i) 'typical', (ii) 'typical with posterior extension', (iii) 'bilateral' and (iv) 'atypical' patterns. Outcome (minimum of 2 years follow-up) was classified as either seizure free, or not seizure free. Actuarial analysis was used to test the relationship of SPECT patterns with outcome. There were 35 cases with the typical ictal SPECT pattern, 13 posterior, nine bilateral and six atypical cases. The atypical pattern was associated with lack of pathology in the surgical specimen. Outcome was similar for the typical, posterior and bilateral with 60%, 69% and 67% seizure free, respectively. In contrast, the atypical group had a worse outcome with only 33% seizure free. Actuarial analysis showed a significant difference in outcome between patients with the typical pattern, and patients with the atypical pattern (P = 0.04). We conclude that extended patterns of ictal perfusion in temporal lobe epilepsy do not predict poor outcome, indicating that extended hyperperfusion probably represents seizure propagation pathways rather than intrinsically epileptogenic tissue. Atypical patterns of hyperperfusion are associated with poor outcome and may indicate diffuse or extra temporal epileptogenicity. PMID- 9397012 TI - Results of surgical treatment in temporal lobe epilepsy with chronic psychosis. AB - The combination of psychosis and refractory temporal lobe epilepsy is not rare. However, patients with chronic interictal psychosis and refractory epilepsy are rejected from many epilepsy surgery programmes purely on psychiatric grounds. It is often assumed that disturbed behaviour will prevent adequate preoperative evaluation or that the patients are unable to provide informed consent for preoperative investigations and for surgery. The observation that the psychosis usually does not improve after operation and fears of an exacerbation of psychosis with post-surgical seizure remission, analogous to 'forced normalization', are further deterrents to surgery in these patients. We describe five patients with the dual diagnoses of medically intractable temporal lobe epilepsy and chronic psychosis who underwent temporal lobe resection. The patients were able to provide informed consent and were easily managed during preoperative investigation. Seizure outcome has been excellent in all. Neither temporal lobe resection nor remission of seizures influenced the nature or evolution of the psychosis. Subjectively the patients functioned better in activities of daily living and freedom from seizures improved integration into psychiatric treatment facilities. With appropriate psychiatric intervention, patients with chronic psychosis and refractory epilepsy can participate in presurgical investigation successfully, and can undergo surgery uneventfully. PMID- 9397013 TI - Human hippocampal AMPA and NMDA mRNA levels in temporal lobe epilepsy patients. AB - This study was designed to determine whether hippocampal neuronal AMPA (alpha amino-3-hydroxy-5-methylisoxazole-4-propionic acid) and NMDA (N-methyl-D aspartate) mRNA levels were differentially increased in temporal lobe epilepsy patients compared with those measured in control tissue from non-seizure autopsies. Hippocampi from hippocampal sclerosis patients (n = 28) and temporal mass lesion cases (n = 12) were compared with those from the autopsies (n = 4), and studied for AMPA GluR1-3 and NMDAR1-2 mRNAs using semi-quantitative in situ hybridization, along with fascia dentata and Ammon's horn neuron densities. Compared with the autopsies, and without correction for neuron counts, the mass lesion cases with neuron densities similar to autopsies showed: (i) significantly increased NMDAR2 hybridization densities for fascia dentata granule cells; (ii) increased AMPA GluR3 mRNA densities for Ammon's horn pyramids; and (iii) similar or numerically increased mRNAs for all other subunits and hippocampal subfields. Compared with the autopsies, hippocampal sclerosis cases with decreased neuron densities showed: (i) significantly decreased AMPA GluR1-2 and NMDAR1-2 hybridization densities for Ammon's horn pyramids and (ii) similar or numerically decreased mRNAs for all other subunits and subfields. However, correcting for changes in neuron densities showed that hippocampal sclerosis patients had increased AMPA and NMDA mRNA levels per neuron compared with autopsies, and in the CA2 resistant sector GluR2 mRNA levels were numerically greater than autopsies and mass lesion cases. Furthermore, relative to autopsies both sclerosis and mass lesion hippocampi showed that, in the stratum granulosum, the greatest mRNA increases were in AMPA GluR1 and NMDAR2 compared with the other mRNAs. In chronic temporal lobe seizure patients these results indicate that mass lesion and sclerosis cases show differential increases in hippocampal AMPA and NMDA mRNA levels per neuron compared with autopsies, especially for AMPA GluR1 and NMDAR2 in fascia dentata granule cells. These findings support the hypothesis that temporal lobe seizures are associated with increased ionotropic glutamate receptor mRNA levels and alterations in receptor subunit composition that probably contribute to neuronal hyperexcitability, synchronization and seizure generation. PMID- 9397015 TI - Clinical and neurophysiological features of tick paralysis. AB - The clinical and neurophysiological findings in six Australian children with generalized tick paralysis are described. Paralysis is usually caused by the mature female of the species Ixodes holocyclus. It most frequently occurs in the spring and summer months but can be seen at any time of year. Children aged 1-5 years are most commonly affected. The tick is usually found in the scalp, often behind the ear. The typical presentation is a prodrome followed by the development of an unsteady gait, and then ascending, symmetrical, flaccid paralysis. Early cranial nerve involvement is a feature, particularly the presence of both internal and external ophthalmoplegia. In contrast to the experience with North American ticks, worsening of paralysis in the 24-48 h following tick removal is common and the child must be carefully observed over this period. Death from respiratory failure was relatively common in the first half of the century and tick paralysis remains a potentially fatal condition. Respiratory support may be required for > 1 week but full recovery occurs. This is slow with several weeks passing before the child can walk unaided. Anti-toxin has a role in the treatment of seriously ill children but there is a high incidence of acute allergy and serum sickness. Neurophysiological studies reveal low-amplitude compound muscle action potentials with normal motor conduction velocities, normal sensory studies and normal response to repetitive stimulation. The biochemical structure of the toxin of I. holocyclus has not been fully characterized but there are many clinical, neurophysiological and experimental similarities to botulinum toxin. PMID- 9397014 TI - Cortical grey matter and benzodiazepine receptors in malformations of cortical development. A voxel-based comparison of structural and functional imaging data. AB - Using [11C]flumazenil-PET and statistical parametric mapping (SPM), we have shown recently that regions of increased and decreased benzodiazepine receptor density may be seen in patients with localization-related epilepsy due to malformations of cortical development. These abnormalities were seen both within and beyond lesions visually apparent on high-resolution MRI. We have also shown, using an interactive anatomical segmentation technique and volume-of-interest measurements, that subtle and unsuspected abnormalities of cortical grey matter volume were found in the same group of patients on high-resolution MRI, beyond the lesions visually apparent. In 10 patients with localization-related epilepsy and malformations of cortical development, we have now applied the automated and objective technique of SPM to the analysis of high-resolution structural MRI. Each individual patient was compared with 16 normal control subjects. We have then simultaneously compared the structural and functional data obtained for each individual patient with normal control high-resolution MRI and [11C]flumazenil PET image using a novel technique. This comparison allowed the detection of functional abnormalities that were not accounted for by either visible or unsuspected structural abnormalities, in an automated and statistically rigorous manner. Five patients had abnormalities of cortical grey matter volume detected using SPM; only these five patients had been found abnormal using the previous volume-of-interest technique. Six of the 10 patients showed regions of cerebral cortex with disproportionate flumazenil binding compared with local grey matter volume. This included regions not found to have abnormal flumazenil binding on analysis of the PET data alone. Furthermore, regions found to have abnormal binding on examination of the PET data alone were, in some instances, shown to be accounted for by abnormalities of cortical grey matter volume. We conclude that the analysis of ligand PET data should always include a comparison with structural MRI; such comparisons are greatly facilitated by the novel approach described. PMID- 9397016 TI - Miyoshi-type distal muscular dystrophy. Clinical spectrum in 24 Dutch patients. AB - Miyoshi-type distal muscular dystrophy has now been found to be more frequent outside Japan than was previously thought. We studied 24 Dutch patients with Miyoshi-type distal muscular dystrophy and focused on its clinical expression and natural history, muscle CT-scans and muscle biopsy findings. Our study shows that Miyoshi myopathy is a heterogeneous, slowly progressive disorder. The disease starts with weakness and atrophy of the calves and progressively involves the proximal leg and hip muscles and, in a later stage the shoulder and upper arm muscles. After 10 years disease duration, one-third of the patients are dependent on wheelchairs for out-of-door transportation. Disease progression is related to disease duration and not to early age of onset of symptoms. Onset may be at any age and is asymmetrical in roughly half of the cases. Four cases had been initially diagnosed as idiopathic hyper-CK-aemia. PMID- 9397017 TI - A PET study of voluntary movement in schizophrenic patients experiencing passivity phenomena (delusions of alien control). AB - Schizophrenic patients experiencing passivity phenomena believe their thoughts and actions to be those of external, or alien, entities. We wished to test the hypothesis that voluntary motor action in such patients would be associated with aberrant patterns of activation within the cerebral motor system. We used H2(15)O PET to study patients while they performed paced joystick movements on two occasions 4-6 weeks apart. During the first scan passivity symptoms were maximal, while by the second scan these symptoms had significantly improved in five of the seven patients. Two control groups were also scanned on two occasions: deluded schizophrenic patients without passivity phenomena and normal subjects. In normal subjects, performance of freely selected joystick movements with the right hand, compared with rest, revealed relative activation of prefrontal, premotor, motor and parietal cortical regions. Schizophrenic patients with passivity showed hyperactivation of parietal and cingulate cortices. This hyperactivation remitted in those subjects in whom passivity decreased over time. This reversible hyperactivity was not a feature of schizophrenics without passivity. Given that these hyperactive cerebral regions subserve attention to internal and external bodily space, and the attribution of significance to sensory information, they provide a plausible anatomical substrate for the misattribution of internally generated acts to external entities: the cardinal feature of delusions of passivity (alien control). PMID- 9397019 TI - Impaired recognition of disgust in Huntington's disease gene carriers. AB - Face processing and facial expression recognition were investigated in the earliest stages of Huntington's disease, by studying 40 people who presented for genetic testing. Twenty-three of these 'at risk' individuals turned out not to carry the gene for Huntington's disease (the AR- group). Seventeen were found to be gene carriers (the AR+ group); 15 from genetic testing, and two who showed signs of early stages of Huntington's disease. A number of standard tasks were used to provide background information, including recognition memory for words, picture naming, verbal fluency, and figure copying; none revealed significant differences between the AR+ and AR- groups. Face processing abilities were investigated using tests of identification of familiar (famous) faces, unfamiliar face matching, recognition memory for faces, and recognition of facial expressions of emotion. No statistically significant differences between the AR+ and AR- groups were found for any of these tests, but the AR+ group showed a borderline overall impairment in recognizing facial expressions of emotion (0.05 < P < 0.1). When recognition of each of the six basic emotions used was examined separately, only disgust was found to be significantly impaired. This highly selective deficit in the recognition of disgust was confirmed in the subgroup of 15 individuals shown by genetic testing to be Huntington's gene carriers; it was therefore found in people who were free from clinical symptoms and did not perform significantly more poorly than non-carriers on any of the background tests, on any of the other face processing tasks, and even for recognition of any other basic emotion. This points strongly to the importance of the basal ganglia in the emotion of disgust. PMID- 9397018 TI - Space-based and object-based visual attention: shared and specific neural domains. AB - Visual attention can be primarily allocated to either where an object is in space (with little emphasis on the structure of the object itself) or to the structure of the object (with little emphasis on where in space the object is located). Using PET measures of regional cerebral blood flow (rCBF) to index neural activity, we investigated the shared and specific functional anatomy underlying both of these types of visual attention in a controlled non-cueing non-blocked paradigm that involved identical stimuli across the conditions of interest. The interaction of eye movements with these attentional systems was studied by introducing fixation or free vision as an additional factor. Relative to the control condition, object-based and space-based attention showed significant activations of the left and right medial superior parietal cortex and the left lateral inferior parietal cortex, the left prefrontal cortex and the cerebellar vermis. Significant differential activations were observed during object-based attention in the left striate and prestriate cortex. Space-based attention activated the right prefrontal cortex and the right inferior temporal-occipital cortex. Differential neural activity due to free vision or fixation was observed in occipital areas only. Significant interactions of free vision/fixation on activations due to object-based and space-based attention were observed in the right medial superior parietal cortex and left lateral inferior parietal cortex, respectively. The study provides direct evidence for the importance of the parietal cortex in the control of object-based and space-based visual attention. The results show that object-based and space-based attention share common neural mechanisms in the parietal lobes, in addition to task specific mechanisms in early visual processing areas of temporal and occipital cortices. PMID- 9397020 TI - Working memory impairment in early multiple sclerosis. Evidence from an event related potential study of patients with clinically isolated myelopathy. AB - Auditory and visual event-related potentials were recorded during a short-term memory task in 24 patients who had recently presented with symptomatically and clinically isolated spinal cord syndromes suspected to be due to multiple sclerosis, and in 24 matched control subjects. Event-related potentials (ERPs) were recorded during two sequential components of the working memory task, first the temporary active memorizing of sets of digits and secondly, their subsequent manipulation, namely digit-probe recognition and matching. The patients' reaction times were slower and showed larger increments than those of the control subjects as the number of items to be memorized was increased. The patients' ERPs during both memorizing and probe matching/recognition phases differed significantly from control subjects for both auditory and visual presentations. The more marked changes were seen in a subgroup of eight patients who had the lowest levels of performance on a battery of general tests of memory and who also made significantly more errors in the working memory task as the memory load increased. In this subgroup, the abnormalities of the ERPs during recognition and matching tests occurred in the component of the response that has been shown to be sensitive to memory loading in healthy control subjects. This study provides objective evidence of subclinical working memory dysfunction in patients at an early stage of demyelinating disease, i.e. when they first present with clinically isolated spinal cord lesions and before they have developed symptoms of cognitive or memory dysfunction. The defect at this early stage is either restricted to processes involved in the formation of a memory trace or, more probably, involves both trace formation and the mechanisms that underly recognition ('retrieval') and matching of memory traces in working memory. PMID- 9397021 TI - Comparison of MRI criteria at first presentation to predict conversion to clinically definite multiple sclerosis. AB - We compared MRI criteria used to predict conversion of suspected multiple sclerosis to clinically definite multiple sclerosis. Seventy-four patients with clinically isolated neurological symptoms suggestive of multiple sclerosis were studied with MRI. Logistic regression analysis was used to remove redundant information, and a diagnostic model was built after each MRI parameter was dichotomized according to maximum accuracy using receiver operating characteristic analysis. Clinically definite multiple sclerosis developed in 33 patients (prevalence 45%). The optimum cut-off point (number of lesions) was one for most MRI criteria (including gadolinium-enhancement and juxta-cortical lesions), but three for periventricular lesions, and nine for the total number of T2-lesions. Only gadolinium-enhancement and juxta-cortical lesions provided independent information. A final model which, in addition, included infratentorial and periventricular lesions, had an accuracy of 80%, and having more abnormal criteria, predicted conversion to clinically definite multiple sclerosis strongly. The model performed better than the criteria of Paty et al. (Neurology 1988; 38: 180-5) and of Fazekas et al. (Neurology 1988; 38: 1822-5). We concluded that a four-parameter dichotomized MRI model including gadolinium enhancement, juxtacortical, infratentorial and periventricular lesions best predicts conversion to clinically definite multiple sclerosis. PMID- 9397022 TI - Direct observation of myelination in vivo in the mature human central nervous system. A model for the behaviour of oligodendrocyte progenitors and their progeny. AB - We studied patches of CNS myelin in human retina in vivo to determine the pattern of myelination and the local influence of axons. We analysed the position, area and thickness of the nerve-fibre layer in 60 patches of retinal myelin in 47 eyes (in 37 adults and two adolescents). Five patches in four eyes were studied serially over 6-11 years. Nerve-fibre layer thickness was obtained from an atlas of primate retina, and volumes of myelinated tissue were then estimated for each patch. Retinal myelination occurred in three patterns: thick patches contiguous with the optic disc (type I); thin, striated patches detached from the disc (type II); or massive myelination of the posterior pole associated with severe amblyopia (type III). The papillomacular bundle did not myelinate in types I and II and was relatively spared in type III patches, suggesting that migratory oligodendrocyte progenitors are not supported by these axons. The local nerve fibre layer determined patch size, and quantal myelination was evident with modal peaks of patch volume at 0.16 and 0.64 mm3. Myelination advanced at patch edges when observed over time, consistent with the hypothesis that new oligodendrocytes are produced in adulthood. We propose a theoretical model where patches of retinal myelination are the clonal progeny of a few oligodendrocyte progenitors exhibiting two different behaviours. First, a highly migratory, nonmyelinating progenitor uses larger, phylogenetically older axons as the substrate for movement. Secondly, a more mature progenitor generates myelinating oligodendrocytes well into adult life, but traverses only short distances. Using this data, we can estimate the number of oligodendrocytes in these clones and population doubling-time. This study supports a role for axon-derived signals in the regulation of human oligodendrocyte progenitor behaviour and myelination in vivo. PMID- 9397023 TI - Granulocyte-macrophage colony-stimulating factor crosses the blood--brain and blood--spinal cord barriers. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF), a glycoprotein with hormonal properties, is produced by several cell types, most of which exist outside the CNS. GM-CSF, however, affects the CNS. If capable of crossing from blood to CNS, GM-CSF might be an important signalling molecule between the CNS and periphery. We used an established in vivo method in mice and rats to study passage of radioactively labelled GM-CSF from blood to CNS. We found that GM-CSF crossed the blood-brain barrier and blood-spinal cord barrier significantly faster than the control substance, albumin. Labelled GM-CSF was recovered in intact form by high performance liquid chromatography from brain after peripheral injection, and passage was not significantly reduced by simultaneous injection of unlabelled L-tryptophan. Both findings indicate that the observed passage of radioactivity was intact protein. Capillary depletion experiments showed that most of the GM-CSF was deposited in brain parenchyma rather than cerebral capillary endothelium. Co-injection of unlabelled GM-CSF significantly reduced the passage rate of labelled cytokine across the blood-brain and blood-spinal cord barriers, demonstrating that passage was mediated by a saturable system. In summary, a saturable mechanism transports GM-CSF intact from blood to CNS. PMID- 9397024 TI - Stability of reach-to-grasp movement patterns in Parkinson's disease. AB - The performance of patients with Parkinson's disease on two reach-to-grasp tasks was compared with that of age-matched control subjects. The aim of the study was to determine whether Parkinson's disease patients have problems coordinating concurrently executed tasks within the same system of effectors in a natural context and whether such problems would be exacerbated by increases in task difficulty. We examined how subjects concurrently executed the transport and grasp components of reach-to-grasp movements in the presence of two types of change in task demands: (i) increases in demands for accurate digit pad placement and (ii) use of two reach-to-grasp tasks, i.e. the standard unimanual task and a bimanual task which increased the control and coordination demands relative to the unimanual task. If Parkinson's disease patients have coordination problems they should demonstrate increased impairment with increasing accuracy demands and in the bimanual task; any such differences should be absent or much smaller in the control group. The Parkinson's disease group showed substantial impairments in all conditions, moving about 30% slower than the control group, with much increased jerking and with signs of difficulty controlling the speed of movement. However, there were no consistent indications that the Parkinson's disease group were differentially impaired on the bimanual task nor that movement deficits increased with increasing accuracy requirements. Grasp and transport components were coordinated similarly by Parkinson's disease and control groups in both reach-to-grasp tasks, and the Parkinson's disease group co-ordinated the two limbs in the bimanual task effectively and in a fashion similar to that of the control group. These results are interpreted to mean that higher levels (effector independent levels) of motor programming are preserved in Parkinson's disease and that execution of a motor programme need not be compromised by increasing the number of muscle-/joint-level degrees of freedom which are used. PMID- 9397025 TI - Hereditary demyelinating neuropathy of infancy: a genetically complex syndrome. PMID- 9397026 TI - Transient middle cerebral artery occlusion by intraluminal suture: I. Three dimensional autoradiographic image-analysis of local cerebral glucose metabolism blood flow interrelationships during ischemia and early recirculation. AB - Using autoradiographic image-averaging strategies, we studied the relationship between local glucose utilization (LCMRglc) and blood flow (LCBF) in a highly reproducible model of transient (2-hour) middle cerebral artery occlusion (MCAO) produced in Sprague-Dawley rats by insertion of an intraluminal suture coated with poly-L-lysine. Neurobehavioral examination at 60 minutes after occlusion substantiated a high-grade deficit in all animals. In two subgroups, LCBF was measured with 14C-iodoantipyrine at either 1.5 hours of MCAO, or at 1 hour of recirculation after suture removal. In two other matched subgroups, LCMRglc was measured with 14C-2-deoxyglucose at 1.5 to 2.25 hours of MCAO, and at 0.75 to 1.5 hours of recirculation after 2 hours of MCAO. Average image data sets were generated for LCBF, LCMRglc, and the LCMRglc/LCBF ratio for each study time. Middle cerebral artery occlusion for 2 hours induced graded LCBF decrements affecting ipsilateral cortical and basal ganglionic regions. After 1 hour of recirculation, LCBF in previously ischemic neocortical regions increased by 40% to 200% above ischemic levels, but remained depressed, on average, at about 40% of control. By contrast, frank hyperemia was noted in the previously ischemic caudoputamen. Mean cortical LCBF values during MCAO correlated highly with their respective LCBF values after 1 hour of recirculation (R = 0.93), suggesting that post-ischemic LCBF recovery is related to the depth of ischemia. Despite focal ischemia, LCMRglc during approximately 2 hours of MCAO was preserved, on average, at near-normal levels; but following approximately 1 h of recirculation, LCMRglc became markedly depressed (on average, 55% of control in previously densely ischemic cortical regions). Regression analysis indicated that this depressed glucose utilization was determined largely by the intensity of antecedent ischemia. By pixel analysis, the ischemic core (defined as LCBF 0% to 20% of control) comprised 33% of the ischemic hemisphere, and the penumbra (LCBF 20% to 40%) accounted for 26%. The penumbra was concentrated at the coronal poles of the ischemic lesion and formed a thin shell around the central ischemic core. During 2 hours of MCAO, the LCMRglc/LCBF ratio within the ischemic penumbra was increased four-fold above normal (average, 179 umol/100 mL). In marked contrast, after approximately 1 h recirculation, this uncoupling had almost completely subsided. The companion study (Zhao et al., 1997) further analyzes these findings in relation to patterns of infarctive histopathology. PMID- 9397027 TI - Transient middle cerebral artery occlusion by intraluminal suture: II. Neurological deficits, and pixel-based correlation of histopathology with local blood flow and glucose utilization. AB - We conducted a pixel-based analysis of the acute hemodynamic and metabolic determinants of infarctive histopathology in a reproducible model of temporary (2 hour) middle cerebral artery occlusion (MCAO) produced in rats by an intraluminal suture. Three-dimensional averaged image data sets of local cerebral blood flow (LCBF) and glucose utilization (LCMRglc) acquired in the companion study (Belayev et al., 1997) either at the end of a 2-hour period of MCAO or after 1 hour of recirculation were comapped (using digitized atlas-templates) with data sets depicting the frequency of histological infarction in a matched animal group (n = 8) in which 2 hours of MCAO was followed by 3-day survival, sequential neuro behavioral examinations, and perfusion-fixation and paraffin-embedding of brains for light-microscopic analysis. All rats developed marked postural-reflex and forelimb-placing deficits at 60 minutes of MCAO, signifying high-grade ischemia. Tactile placing deficits persisted during the 72-hour observation period while visual placing and postural-reflex abnormalities variably improved. Comapping of LCBF and histopathology showed that in those pixels destined to undergo infarction, LCBF measured at 2 hours of MCAO showed a sharp distributional peak centered at 0.14 mL/g/min. In 70% of pixels destined to infarct, LCBF at 2 hours of MCAO was 0.24 mL/g/min or below, and in 89% LCBF was below 0.47 mL/g/min (the upper limits of the ischemic core and penumbra, respectively, as defined in the companion study [Belayev et al., 1997]). Local cerebral glucose utilization measured at approximately 1 hour after 2 hours of MCAO was distributed bimodally in the previously ischemic hemisphere. The major peak, at 22 mumol/100g/min, coincided exactly with the distribution peak of pixels destined to undergo infarction, while in pixels with a zero probability of infarction, LCMRglc was higher by 12 to 13 mumol/100g/min. These results indicate that local blood flow at 2 hours of MCAO is a robust predictor of eventual infarction. Pixels with ischemic-core levels of LCBF (0% to 20% of control) have a 96% probability of infarction, while the fate of the penumbra is more heterogeneous: below LCBF of 0.35 mL/g/min, the probability of infarction is 92%, while approximately 20% pixels in the upper-penumbral LCBF range (30% to 40% of control) escape infarction. Our data strongly support the view that the likelihood of infarction within the ischemic penumbra is highly influenced by very subtle differences in early perfusion. PMID- 9397028 TI - Effect of brain ischemia and reperfusion on the localization of phosphorylated eukaryotic initiation factor 2 alpha. AB - Postischemic brain reperfusion is associated with a substantial and long-lasting reduction of protein synthesis in selectively vulnerable neurons. Because the overall translation initiation rate is typically regulated by altering the phosphorylation of serine 51 on the alpha-subunit of eukaryotic initiation factor 2 (eIF-2 alpha), we used an antibody specific to phosphorylated eIF-2 alpha [eIF 2(alpha P)] to study the regional and cellular distribution of eIF-2(alpha P) in normal, ischemic, and reperfused rat brains. Western blots of brain postmitochondrial supernatants revealed that approximately 1% of all eIF-2 alpha is phosphorylated in controls, eIF-2(alpha P) is not reduced by up to 30 minutes of ischemia, and eIF-2(alpha P) is increased approximately 20-fold after 10 and 90 minutes of reperfusion. Immunohistochemistry shows localization of eIF-2(alpha P) to astrocytes in normal brains, a massive increase in eIF-2(alpha P) in the cytoplasm of neurons within the first 10 minutes of reperfusion, accumulation of eIF-2(alpha P) in the nuclei of selectively vulnerable neurons after 1 hour of reperfusion, and morphology suggesting pyknosis or apoptosis in neuronal nuclei that continue to display eIF-2(alpha P) after 4 hours of reperfusion. These observations, together with the fact that eIF-2(alpha P) inhibits translation initiation, make a compelling case that eIF-2(alpha P) is responsible for reperfusion-induced inhibition of protein synthesis in vulnerable neurons. PMID- 9397029 TI - Hyperglycemia and hypercapnia suppress BDNF gene expression in vulnerable regions after transient forebrain ischemia in the rat. AB - Preischemic hyperglycemia or superimposed hypercapnia exaggerates brain damage caused by transient forebrain ischemia. Because high regional levels of brain derived neurotrophic factor (BDNF) protein correlate with resistance to ischemic damage, we studied the expression of BDNF mRNA using in situ hybridization in rats subjected to 10 minutes of forebrain ischemia under normoglycemic, hyperglycemic, or hypercapnic conditions. Compared with normoglycemic animals, the increase of BDNF mRNA using in situ hybridization in rats subjected to 10 minutes of forebrain ischemia under normoglycemic, or hypercapnic conditions. Compared with normoglycemic animals, the increase of BDNF mRNA in dentate granule cells was attenuated and that in CA3 pyramidal neurons completely prevented in hyperglycemic rats. No ischemia-induced increases of BDNF mRNA levels in the hippocampal formation were detected in hypercapnic animals. Hyperglycemic and hypercapnic rats showed transiently decreased expression of BDNF mRNA levels in the cingulate cortex, which was not observed in normoglycemic animals. The results suggest that suppression of the BDNF gene might contribute to the increased vulnerability of the CA3 region and cingulate cortex in hyperglycemic and hypercapnic animals. PMID- 9397030 TI - Blockade of cerebral blood flow response to insulin-induced hypoglycemia by caffeine and glibenclamide in conscious rats. AB - The possibility that adenosine and ATP-sensitive potassium channels (KATP) might be involved in the mechanisms of the increases in cerebral blood flow (CBF) that occur in insulin-induced hypoglycemia was examined. Cerebral blood flow was measured by the [14C]iodoantipyrine method in conscious rats during insulin induced, moderate hypoglycemia (2 to 3 mmol/L glucose in arterial plasma) after intravenous injections of 10 to 20 mg/kg of caffeine, an adenosine receptor antagonist, or intracisternal infusion of 1 to 2 mumol/L glibenclamide, a KATP channel inhibitor. Cerebral blood flow was also measured in corresponding normoglycemic and drug-free control groups. Cerebral blood flow was 51% higher in untreated hypoglycemic than in untreated normoglycemic rats (P < 0.01). Caffeine had a small, statistically insignificant effect on CBF in normoglycemic rats, but reduced the CBF response to hypoglycemia in a dose-dependent manner, i.e., 27% increase with 10 mg/kg and complete elimination with 20 mg/kg. Chemical determinations by HPLC in extracts of freeze-blown brains showed significant increases in the levels of adenosine and its degradation products, inosine and hypoxanthine, during hypoglycemia (P < 0.05). Intracisternal glibenclamide had little effect on CBF in normoglycemia, but, like caffeine, produced dose dependent reductions in the magnitude of the increases in CBF during hypoglycemia, i.e., +66% with glibenclamide-free artificial CSF administration, +25% with 1 mumol/L glibenclamide, and almost complete blockade (+5%) with 2 mumol/L glibenclamide. These results suggest that adenosine and KATP channels may play a role in the increases in CBF during hypoglycemia. PMID- 9397031 TI - Cerebral blood flow during hypoxemia and hemodilution in rabbits: different roles for nitric oxide? AB - Hypoxemia and anemia are associated with increased CBF, but the mechanisms that link the changes in PaO2 or arterial O2 content (CaO2) with CBF are unclear. These experiments were intended to examine the contribution of nitric oxide. CaO2 in pentobarbital-anesthetized rabbits was reduced to approximately 6.5 mL O2/dL by hypoxemia (PaO2 approximately 24 to 26 mm Hg) or hemodilution with hetastarch (hematocrit approximately 14% to 15%). Animals with normal CaO2 (approximately 17.5 to 18 mL O2/dL) served as controls. In part I, each animal was given 3, 10, and 30 mg/kg N omega-nitro-L-arginine methyl ester (L-NAME) intravenously (total 43 mg/kg) to inhibit production of nitric oxide. Forebrain CBF was measured with radioactive microspheres approximately 15 to 20 minutes after each dose. Baseline CBF was greater in hypoxemic rabbits (111 +/- 31 mL x 100 g-1 x min-1, mean +/- SD) than in hemodiluted (70 +/- 22 mL x 100 g-1 min-1) or control animals (39 +/- 12 mL x 100 g-1 min-1). L-NAME (which reduced brain tissue nitric oxide synthase activity by approximately 65%) reduced CBF in hypoxemic animals to 80 +/- 23 mL x 100 g-1 x min-1 (P < 0.0001), but had no significant effect on CBF in either anemic or control animals. In four additional rabbits, further hemodilution to a CaO2 of approximately 3.5 mL O2/dL increased baseline CBF to 126 +/- 21 mL x 100 g-1 min-1, but again there was no effect of L-NAME. In part II, animals were anesthetized as above, and a close cranial window was prepared. The cyclic GMP (cGMP) content of the artificial CSF superfusate was measured under baseline conditions, and then after the reduction of CaO2 to approximately 6.5 mL O2/dL by either hypoxemia or hemodilution. Concentrations of cGMP did not change during either control conditions or after hemodilution. However, cGMP increased significantly with the induction of hypoxemia. The cGMP increase in hypoxemic animals could be blocked with L-NAME. These results suggest that nitric oxide plays some role in hypoxemic vasodilation, but not during hemodilution. PMID- 9397032 TI - Laminar analysis of cerebral blood flow in cortex of rats by laser-Doppler flowmetry: a pilot study. AB - Laser-Doppler flowmetry (LDF) is a reliable method for estimation of relative changes of CBF. The measurement depth depends on wavelength of the laser light and the separation distance of transmitting and recording optical fibers. We designed an LDF probe using two wavelengths of laser light (543 nm and 780 nm), and three separation distances of optical fibers to measure CBF in four layers of the cerebral cortex at the same time. In vitro comparison with electromagnetic flow measurements showed linear relationship between LDF and blood flow velocity at four depths within the range relevant to physiologic measurements. Using artificial brain tissue slices we showed that the signal for each channel decreased in a theoretically predictable fashion as a function of slice thickness. Application of adenosine at various depths in neocortex of halothane anesthetized rats showed a predominant CBF increase at the level of application. Electrical stimulation at the surface of the cerebellar cortex demonstrated superficial predominance of increased CBF as predicted from the distribution of neuronal activity. In the cerebellum, hypercapnia increased CBF in a heterogeneous fashion, the major increase being at apparent depths of approximately 300 and 600 microns, whereas in the cerebral cortex, hypercapnia induced a uniform increase. In contrast, the CBF response to cortical spreading depression in the cerebral cortex was markedly heterogeneous. Thus, real-time laminar analysis of CBF with spatial resolution of 200 to 300 microns may be achieved by LDF. The real-time in depth resolution may give insight into the functional organization of the cortical microcirculation and adaptive features of CBF regulation in response to physiologic and pathophysiologic stimuli. PMID- 9397033 TI - Vasodilator effects on canine basilar artery induced by intracisternal interleukin-1 beta. AB - The effect of interleukin-1 beta (IL-1 beta) on a cerebral artery was investigated in anesthetized dogs. Intracisternal administration of IL-1 beta (0.03 and 0.3 micrograms) dilated the canine basilar artery in a dose-dependent manner, without affecting systemic blood pressure or heart rate. The increase in diameter induced by 0.3 micrograms of IL-1 beta was 28.4% +/- 13.4% of control at 2 hours and was inhibited by 30 micrograms of the IL-1 beta receptor antagonist, zinc protoporphyrin (4.5% +/- 13.5%, P < 0.05). Interleukin-1 beta did not affect the concentration of nitric oxide metabolites in CSF. However, there was an increase in the concentration of eicosanoids in CSF, and the elevation of 6-keto PGF1 alpha paralleled the vasodilation. Pretreatment with 30 micrograms of the selective inducible cyclooxygenase (COX-2) inhibitor NS-398 also inhibited the IL 1 beta-induced vasodilation significantly (5.9% +/- 9.4% at 2 hours, P < 0.01). Western blot analysis revealed the expression of a 68-kD COX-2-like protein in basilar artery extracts. These findings suggest that the IL-1 beta-induced vasodilator effect is linked to the prostaglandin cascade, predominantly to prostaglandin I2, by induction of COX-2, but not to the stimulation of nitric oxide metabolism. PMID- 9397034 TI - Primary prevention of skin cancer: where to now in reducing sunlight exposure? PMID- 9397035 TI - Mammographic screening in Australia. PMID- 9397036 TI - Surgery for severe emphysema. PMID- 9397037 TI - "Single use only": obfuscation or the necessary attainment of zero risk? PMID- 9397038 TI - Mammographic screening: results from the 1996 National Breast Health Survey. AB - OBJECTIVE: To establish the extent of women's knowledge of mammographic screening, particularly in relation to the national screening program, BreastScreen Australia, and to estimate the proportion of women who are participating in screening both within and outside BreastScreen Australia. DESIGN AND SETTING: Validated prospective telephone survey of women aged 30-69 years selected at random from across Australia. PARTICIPANTS: 2935 women with no previous breast cancer diagnosis. RESULTS: The adjusted response rate was 64%. Almost 90% of women had heard of the national program; only 1% correctly stated that screening is for asymptomatic women. 60% correctly identified the current recommended age of starting screening is about 50 years of age; 26% thought screening should begin at about 40 years of age. Approximately 60% correctly reported that the recommended screening interval is every two years; 27% thought screening should be done annually. 55% reported ever having had a mammogram, and 37% reported having had at least one screening mammogram. Among women in the target age group (50-69 years) about 70% reported ever having had a screening mammogram, and about 50% reported having had a screening mammogram within the national program in the last two years. Among women aged 40-49 years, 29% reported ever having had a screening mammogram, and 22% reported having been screened in the last two years. CONCLUSIONS: Awareness of the national screening program is high, but some women do not know the purpose of screening, the target age group and the recommended screening interval. Compliance with screening is good among women in the target age group; many women in their 40s are also participating in screening. PMID- 9397039 TI - Self-reported morbidity of Barmah Forest virus infection on the north coast of New South Wales. AB - OBJECTIVE: To describe the clinical features and disability associated with Barmah Forest virus (BFV) infection. DESIGN: Retrospective postal survey. SETTING: North Coast Public Health Unit, Lismore, New South Wales, January to October 1995. SUBJECTS: All 84 subjects notified by mandatory laboratory reporting as positive for BFV IgM by enzyme-linked immunosorbent assay. OUTCOME MEASURES: Demographic information, self-reported symptoms, disability and treatment. RESULTS: Response rate was 77%. Peak incidence was in the 30-50 years age group, with almost identical numbers of men and women affected. The most common symptoms were lethargy (89%), joint pain (82%) and rash (68%). These were also generally the first symptoms to appear. Thirty of 54 respondents (56%) reported time off work and 27 of 53 (51%) reported illness lasting more than six months. Those who had a rash were significantly more likely to have recovered by the time of the survey than those who had no rash (odds ratio, 10.3; 95% confidence interval, 1.8-76.6). No treatment led to more than slight relief of symptoms. CONCLUSION: Symptoms of BFV infection appear similar to those of the better-known Ross River virus infection, and clinicians should consider both in patients with symptoms of arboviral disease. The wide distribution and long duration of illness make BFV a potentially significant cause of morbidity in Australia. A possible association between the presence of a rash and improved prognosis needs further investigation. PMID- 9397041 TI - Eye injuries caused by elasticated "octopus" straps. AB - "Octopus" elasticated straps are a common cause of severe accidental eye injuries, which we believe are largely preventable. In a retrospective study (January 1990 to August 1996), we identified 42 patients with such injuries severe enough to warrant admission to hospital. The injuries included hyphaema, vitreous haemorrhage, retinal detachment, and choroid and globe rupture, with 28% (12/42) of the injuries resulting in permanent visual loss. We believe octopus straps should not be available for sale in their current form. PMID- 9397040 TI - Lung volume reduction surgery for emphysema. AB - OBJECTIVE: To report the results of lung volume reduction surgery (LVRS) for severe emphysema in Australia. SETTING: A tertiary teaching hospital. DESIGN: A prospective study of a consecutive case series. PARTICIPANTS: 20 patients (mean age, 56 years) with severe emphysema--mean forced expiratory volume in one second (FEV1), 0.72 L (28% of predicted) and severe gas trapping (mean residual volume, 286% of predicted). INTERVENTION: Bilateral apical LVRS was performed via a median sternotomy with a linear stapler; bovine pericardial strips were used to reinforce the staple line. RESULTS: There was a 95% survival, and a mean (range) inpatient stay of 17 (8-45) days. No complications occurred in nine patients; a further six patients had only minor complications. Five patients had major complications (sputum retention requiring reintubation, persistent air leak requiring reoperation, duodenal perforation, and epidural haemorrhage); one patient died from multiorgan failure at 28 days. Intercostal drainage was left in situ for a mean of eight days. The results of FEV1, Medical Research Council (MRC) Dyspnoea Score and six-minute walk test improved in more than 90% of patients. FEV1 improved an average of 0.35 L (54% over baseline) (P < 0.001). Mean MRC Dyspnoea Score decreased from 3.4 to 2.1 (P < 0.001). Mean distance for the six-minute walk test increased from 306 to 431 metres (P < 0.001). CONCLUSION: Our experience confirms that LVRS produces worthwhile early outcomes for a subgroup of patients with severe emphysema. The clinical, economic and ethical questions raised by this new therapy will need to be assessed. PMID- 9397042 TI - "Single use only" labelling of medical devices: always essential or sometimes spurious? PMID- 9397043 TI - Specific allergen immunotherapy for asthma. A position paper of the Thoracic Society of Australia and New Zealand and the Australasian Society of Clinical Immunology and Allergy. PMID- 9397044 TI - MJA practice essentials. 6. Stress management and counselling in primary care. PMID- 9397045 TI - Malnutrition and microcephaly in Australian aboriginal children. PMID- 9397046 TI - The prevalence of hookworm infection, iron deficiency and anaemia in an aboriginal community in north-west Australia. PMID- 9397047 TI - Politicised Aborigine health research. PMID- 9397048 TI - Incompletely excised basal cell carcinomas of the skin. PMID- 9397049 TI - Incompletely excised basal cell carcinomas of the skin. PMID- 9397050 TI - "Best practice" in surgical management of breast cancer. PMID- 9397051 TI - Vaccination--is the evidence good enough to say there is no risk? PMID- 9397052 TI - "On-the-spot" vaccination. PMID- 9397053 TI - Rapid identification of Plasmodium falciparum malaria. PMID- 9397054 TI - Kava and alcohol. PMID- 9397055 TI - Crohn's and colitis: science progresses steadily. PMID- 9397056 TI - Automation in cervical cytology: whose cost and whose benefit? PMID- 9397057 TI - How can we best achieve optimal transfusion practice? PMID- 9397058 TI - Huntington's disease: a challenge for out times. PMID- 9397059 TI - Evaluation of the ThinPrep Pap test as an adjunct to the conventional Pap smear. AB - OBJECTIVE: To evaluate the ThinPrep Pap test as an adjunct to the conventional Pap smear. DESIGN AND SETTING: Prospectively collected cervical samples were split for independent screening at a large specialised private gynaecological pathology practice in Sydney. MAIN OUTCOME MEASURES: Detection of additional significant abnormalities (cervical intraepithelial neoplasia 1, or more severe); changed management recommendations from "repeat smear in 12 months" or "...six months" to "colposcopy", a reduction in unsatisfactory reports. RESULTS: 35,560 paired (split-sample) conventional and ThinPrep slides were prepared. Significant abnormalities were detected in 724 conventional smears (2%). Additional significant abnormalities were found in 85 ThinPrep slides whose corresponding conventional smear was negative or unsatisfactory even after review, representing a 12% increase in the detection of significant abnormalities. As a result of the addition of ThinPrep, management recommendations were changed from "repeat smear in 12 months" or "...six months" to "colposcopy" for 89 of 1669 women whose conventional Pap smears showed minor non-specific changes or papillomavirus. There were 1258 conventional smears (3.5%) that were unsatisfactory compared with 235 ThinPrep slides (0.7%); for only 74 samples (0.2%) were both slides unsatisfactory. CONCLUSIONS: The addition of the ThinPrep Pap test improves detection and clinical management of cervical abnormalities, and reduces the number of unsatisfactory samples which would otherwise require repeat tests. PMID- 9397060 TI - Prevalence of hepatitis C infection in pregnant women in South Australia. AB - OBJECTIVES: To estimate the prevalence of hepatitis C virus (HCV) seropositivity and known risk factors for HCV infection in a group of pregnant women. DESIGN: Cross-sectional survey. SETTING: Lyell McEwin Health Service, Elizabeth, South Australia (a general public hospital with an annual average of about 2000 deliveries). SUBJECTS: 1537 consecutive women who delivered at the Lyell McEwin Health Service from February 1995 to December 1995. OUTCOME MEASURES: Presence of HCV antibodies; and associations between HCV-antibody status and known risk factors. RESULTS: 17 women (1.1%) were HCV-seropositive. Risk factors significantly more prevalent among HCV-seropositive patients were: a history of injecting drug use, a past or present sexual partner who had injected drugs, having a tattoo and having been incarcerated. The proportions who had received a blood transfusion, had acquired a sexually transmitted disease or were positive for hepatitis B virus surface antigen were not significantly different between seropositive and seronegative women. Multivariate analysis showed that only injecting drug use remained a strong independent predictor of HCV-seropositivity (odds ratio [OR], 50.1; P < 0.001), while having a tattoo approached significance (OR, 3.5; P = 0.07). CONCLUSION: As only 1.1% of this sample of women were HCV seropositive, screening of all pregnant women does not seem warranted. Testing on the basis of a history of risk factors, such as injecting drug use and having a tattoo, would detect undiagnosed HCV infections more efficiently. PMID- 9397061 TI - Reduction of inappropriate use of blood products by prospective monitoring of transfusion request forms. AB - OBJECTIVE: To determine the effect of prospective monitoring on appropriateness of transfusions of red cells, platelets and fresh frozen plasma (FFP). DESIGN: Prospective interventional study. SETTING: Royal Melbourne Hospital (a tertiary teaching hospital), Melbourne, Victoria, March-May 1996. INTERVENTION: The blood product request form was modified to incorporate indications for transfusion and clinical and laboratory data. Requests were monitored by blood bank laboratory staff for conformation with hospital transfusion guidelines; non-conforming requests were discussed with the requesting medical practitioner by the Haematology Registrar before blood products were issued. In case of disagreement, blood products were always issued. SUBJECTS: 200 consecutive transfusion episodes for each product (red cells, platelets and FFP). OUTCOME MEASURES: Appropriateness of transfusion, assessed by a Consultant Haematologist according to hospital guidelines. Rates of inappropriate transfusion episodes after intervention were compared with rates in a previous study. RESULTS: After intervention, rates of inappropriate transfusion episodes fell significantly (red cells, 16% to 3% [P = 0.004]; platelets, 13% to 2.5% [P = 0.02]; and FFP, 31% to 15% [P = 0.02]). Almost all inappropriate FFP transfusion episodes post intervention were due to failure to demonstrate prolongation of prothrombin or activated partial thromboplastin times more than 1.5 times the control value. CONCLUSION: Prospective monitoring of request forms can reduce rates of inappropriate transfusions. High rates of inappropriate FFP transfusions possibly reflect uncertainty about appropriate laboratory criteria for FFP transfusion. While results of large prospective randomised controlled clinical trials of FFP transfusions are awaited, currently laboratory criteria can be retained, but should be applied with flexibility. PMID- 9397062 TI - Leptospirosis presenting as a haemorrhagic fever in a traveller from Africa. AB - Leptospirosis is usually a mild illness, although the severity of clinical manifestations may vary between the serovars of leptospires. In May 1993, a 48 year-old man from Ghana presented with severe icteric leptospirosis, initially managed as viral haemorrhagic fever. The causative serovar, bataviae, had not been previously diagnosed in human infection in Australia. PMID- 9397063 TI - The Australian health care system: John Hunter's long shadow. PMID- 9397064 TI - New drugs, old drugs. Calcium antagonists. PMID- 9397065 TI - Benzodiazepines in anxiety disorders: managing therapeutics and dependence. PMID- 9397066 TI - Occupational lung disease. PMID- 9397067 TI - Post-traumatic stress disorder: what's in a name? PMID- 9397068 TI - Dantrolene and "ecstasy". PMID- 9397069 TI - Increasing the accuracy of the Pap test. PMID- 9397070 TI - Preventing brain damage in boxers. PMID- 9397071 TI - Confidentiality and the AMA's new code of ethics. PMID- 9397072 TI - An inexpensive "orthosis" for plantar fasciitis. PMID- 9397073 TI - Dwindling supplies of anti-D. PMID- 9397074 TI - Sexually transmitted diseases and contact tracing. PMID- 9397075 TI - Clinical practice guidelines: to what end? PMID- 9397076 TI - Parents' views on reduced length of stay for their asthmatic children. PMID- 9397077 TI - Reduction in length of hospital stay for acute childhood asthma associated with the introduction of casemix funding. PMID- 9397078 TI - Medical treatment of caustic burns. PMID- 9397079 TI - Peritoneal catheter fracture caused by a seatbelt. PMID- 9397080 TI - Efficacy and tolerability of ketoprofen 200 mg controlled-release cps vs indomethacin 50 mg cps in patients with symptomatic hip osteoarthritis. A multicentre study. AB - BACKGROUND: An open-label, randomised, multicentre study was carried out to compare the efficacy and tolerability of indomethacin capsules and ketoprofen controlled-release capsules in the symptomatic treatment of coxarthrosis. MATERIALS AND METHODS: 113 out-patients were enrolled: 57 were assigned to receive indomethacin 50 mg twice daily and 56 ketoprofen 200 mg once daily for 4 weeks. RESULTS: Indomethacin and ketoprofen proved equally effective in relieving osteoarticular pain and stiffness and in improving the quality of life of patients. There was essentially no difference as to gastrointestinal adverse events which occurred in 25% of patients on indomethacin and in 27% of those on ketoprofen. Indomethacin caused more non-gastrointestinal untoward effects, especially CNS effects (headache and dizziness: 11%) which were not observed with ketoprofen. Indomethacin was discontinued because of adverse events in a larger proportion of patients (20%) than ketoprofen (11%). PMID- 9397081 TI - Occult coeliac disease and iron-deficiency anaemia. AB - Coeliac disease can cause selective malabsorption and, therefore, a poorly specific clinical pattern. In the two cases of iron-deficiency anaemia described, targeted diagnostic procedures enabled to find an effective therapy. The identification of patients affected by coeliac disease in a subclinical phase may reduce the risk of autoimmune and lymphoproliferative diseases. PMID- 9397082 TI - Maori/non-Maori patterns of contact, expressed morbidity and resource use in general practice: data from the Waikato Medical Care Survey 1991-2. AB - AIMS: To compare patterns of contact, expressed morbidity and resource use in primary care for a representative sample of patients of Maori and non-Maori background. METHODS: The data are drawn from a survey of general practice in the Waikato region representing a one per cent sample of all week day encounters. The data were recorded by participating general practitioners in four collection weeks spaced over the period of a year. In total, 12,833 patient encounter forms were completed. RESULTS: Annual rates of general practitioner contact for Maori are slightly lower than those for patients of non-Maori background. The case-mix pattern of general practitioner contact is very similar between the two groups. There is a limited correspondence between ethnic patterns of general practitioner usage and health need (as measured by mortality levels and rates of public hospital discharge). CONCLUSIONS: The near equivalence in ethnic rates of general practitioner contact revealed in this study contrasts strikingly both with the level of hospitalisation for Maori, which is nearly double that of non-Maori, and with the difference in mortality rates (30% higher for Maori). Attention devoted to improving access to general practitioner services among Maori may be necessary if important areas of ill health and hospital resource use are to be addressed effectively. PMID- 9397083 TI - The use of behavioral methods of contraception in women seeking abortion. AB - AIMS: To determine the extent to which behavioural methods, used alone or with other methods of contraception, contribute to contraceptive failure in women seeking termination of pregnancy. METHOD: A clinical audit was conducted of the use of behavioural methods of contraception, in 1342 women attending the Parkview Clinic for termination of pregnancy, over a four year period 1992-6. The information was obtained through standard interview and patient records. RESULTS: The women seen were representative of New Zealand women undergoing termination of pregnancy. Approximately one third of women (34.9%) used one or more behavioural methods in the month prior to conception. The two most common methods were periodic abstinence and coitus interruptus, both used by approximately 17%. Other methods used less frequently and not solely for contraceptive purposes, were cleansing of the vagina and breastfeeding. Pacific Island and Asian women were more likely to use behavioural methods than European and Maori women. Many women (45.6%) using behavioural methods also used other methods of contraception, especially the condom. CONCLUSIONS: Behavioural methods were found more commonly than previously reported. Greater understanding of the use of behavioural methods will assist in the implementation of preventive programmes to reduce the number of terminations. PMID- 9397084 TI - Attitudes to the use of dummies in New Zealand; a qualitative study. AB - AIMS: To explore experiences with, and attitudes to, the use of dummies (pacifiers). METHODS: Seven focus group discussions were held with groups of mothers and of health professionals. RESULTS: Most mothers and health care workers had a generally negative view of dummy use. This related particularly to dislike of toddlers with them and practical issues such as getting lost or dirty. All would allow their use in a very unsettled baby. No mothers had personally experienced problems with breastfeeding due to the use of a dummy, but concern about this possibility was expressed by some health care workers. Recommendations varied about the length of time that dummies need to be avoided. CONCLUSIONS: Mothers in New Zealand use dummies selectively for their infants and were concerned with issues of weaning the baby from the dummy, keeping it clean and not losing it. In analysing the relationships between dummy use and breastfeeding it is important to take into consideration the context of dummy use. PMID- 9397085 TI - Consensus viewpoint on the treatment of postmenopausal osteoporosis. The Ad Hoc Group on Osteoporosis. AB - Treatment for postmenopausal osteoporosis should be offered to those with a history of fractures following minimal trauma or with a bone density significantly below the range seen in young normal adults. Underlying diseases contributing to the reduced bone density should be sought and treated appropriately. Lifestyle issues such as smoking, alcohol intake and exercise should be addressed. A calcium intake of at least 1.5 g/day should be achieved. Hormone replacement therapy is the first line pharmacological intervention. The bisphosphonates provide a satisfactory alternative for those unable or unwilling to take hormone replacement therapy. PMID- 9397086 TI - Changing to injectable polio vaccine. PMID- 9397087 TI - Long acting bronchodilators in chronic obstructive pulmonary disease. PMID- 9397088 TI - Fluvastatin. PMID- 9397089 TI - Fluvastatin. PMID- 9397090 TI - Medical evidence for aegrotats in School Certificate and Bursary examinations. PMID- 9397091 TI - Cyclospora cayetanensis diarrhoea in a traveller. PMID- 9397092 TI - Pregnancy outcomes in women with gestational diabetes compared with the general obstetric population. AB - OBJECTIVE: To compare pregnancy outcome in a homogeneous group of women with glucose intolerance with that of women without this disorder. METHODS: This was a retrospective cohort study of all women with singleton cephalic-presenting pregnancies delivered at University of Texas Southwestern Medical Center during the period January 1, 1991, through December 31, 1995. During this period, women were screened selectively for glucose intolerance and National Diabetes Data Group thresholds were used to diagnose gestational diabetes. Women with class A1 gestational diabetes were compared with nondiabetic women within the cohort. Effects of confounding variables were analyzed using multiple logistic regression and a matched-control comparison. Controls were matched according to ethnicity, maternal age, maternal weight, and parity. RESULTS: A total of 61,209 nondiabetic women with singleton cephalic pregnancies were delivered during the study period, and 874 were diagnosed with class A1 gestational diabetes. Women with class A1 gestational diabetes were significantly older, heavier, of greater parity, and more often of Hispanic ethnicity. Hypertension (17 versus 12%), cesarean delivery (30 versus 17%), and shoulder dystocia (3 versus 1%) were significantly increased (all P < .001) in these women compared with the general obstetric population. Infants born to women with class A1 gestational diabetes were significantly larger (mean birth weight 3581 +/- 616 versus 3290 +/- 546 g, P < .001), and this accounted for the increased incidence of dystocia. The attributable risk for large for gestational age (LGA) infants due to class A1 gestational diabetes was 12%. CONCLUSION: The main consequence of class A1 gestational diabetes is excessive fetal size leading to increased risk of difficult labor and delivery. We estimate that approximately one of eight women with class A1 gestational diabetes mellitus delivers an LGA infant attributable to glucose intolerance. PMID- 9397093 TI - Circulating cell adhesion molecule concentrations in diabetic women during pregnancy. AB - OBJECTIVE: To determine whether circulating concentrations of defined cell adhesion molecules, which are thought to reflect endothelial expression, are increased in insulin-dependent diabetic women during pregnancy. METHODS: Pregnant diabetic women demonstrating good glycemic control and without major complications before pregnancy were studied at 8-12 (n = 15), 18 (n = 15), 28 (n = 16), 32 (n = 16), and 36 (n = 16) weeks' gestation. A subgroup of ten diabetic women was sampled longitudinally through all five gestational ages. The diabetic women were compared with healthy nondiabetic women sampled cross sectionally at 12 (n = 20), 28 (n = 19), and 36 (n = 19) weeks' gestation. Nonpregnant diabetic (n = 22) and nonpregnant nondiabetic women (n = 28) also were studied. Plasma concentrations of the cell adhesion molecules E-selectin, intercellular adhesion molecule-1 (ICAM-1), and vascular endothelial cell adhesion molecule-1 (VCAM-1) were measured by enzyme-linked immunosorbent assay. RESULTS: Significantly higher median (range) concentrations of E-selectin (63.0 [20.2-107.0] ng/mL) and ICAM-1 (281.5 [171.6-778.4] ng/mL) but not VCAM-1 (459.7 [301.0-909.7] ng/mL) were found in nonpregnant diabetic women compared with nonpregnant nondiabetic women (43.5 [18.1-93.2], 243.6 [174.4-329.2], and 476.0 [253.8-929.4] ng/mL, respectively). During pregnancy these significant differences between diabetic and control groups were lost. The median (range) concentration of E-selectin (50.0 [21.2 96.3] ng/mL) was significantly lower in pregnant compared with nonpregnant diabetic women. The plasma concentrations of E-selectin and ICAM-1 did not change significantly with gestation in either diabetic or nondiabetic pregnant groups. Vascular endothelial cell adhesion molecule-1 concentration changed significantly with gestation in the diabetic pregnant group only. CONCLUSION: Circulating concentrations of defined vascular cell adhesion molecules are not increased abnormally in diabetic women with good glycemic control during otherwise uncomplicated pregnancy. PMID- 9397094 TI - Gestational diabetes mellitus in women receiving beta-adrenergics and corticosteroids for threatened preterm delivery. AB - OBJECTIVE: To determine whether the incidence of gestational diabetes mellitus (GDM) is increased in patients receiving corticosteroids with or without beta adrenergic agents for threatened preterm delivery. METHODS: We reviewed the laboratory records of 3396 patients undergoing screening (1-hour glucose) and diagnostic testing (3-hour glucose tolerance test [GTT]) for GDM over 2 years. Patients with antepartum admissions during which they received corticosteroids with or without beta-adrenergic agents for threatened preterm delivery were compared with a control group during the same period. Differences between the study and control groups were analyzed using chi 2, Student t test, or Fisher exact test where appropriate. P < .05 was considered significant. RESULTS: Fifty patients in the study group were compared with 1985 control patients. The remaining 1361 patients failed to meet inclusion criteria. The overall incidence of diagnosed GDM was significantly greater in the corticosteroid-beta-adrenergic agents study group, in which five (23.8%) of 21 patients screened had abnormal 3 hour GTT results, compared with 79 (4.0%) of 1985 controls (P = .001). One-hour glucose screening test results were abnormal in 60% of the study group compared with 25% of the controls (P < .001). CONCLUSION: Patients treated with beta adrenergic agents and corticosteroids for threatened preterm delivery are at a significantly increased risk for developing GDM. The high rate of abnormal results in response to the 1-hour glucose screen suggests that this test is of limited value in patients exposed to these medications. PMID- 9397095 TI - Risk of preeclampsia in second-trimester triploid pregnancies. AB - OBJECTIVE: To determine the magnitude of the risk and the predictive clinical characteristics for development of preeclampsia when triploidy is diagnosed in the second trimester. METHODS: A retrospective analysis of databases maintained by the cytogenetics laboratories at the University of Iowa and University of North Carolina was performed to identify all cases of triploidy. We examined the karyotype, maternal serum screening (particularly the hCG level), ultrasound results, and evidence of maternal hypertensive disease. RESULTS: Seventeen cases of triploidy were identified between 1987 and 1996. Preeclampsia or hypertension complicated six of these cases with onset between 15 and 22.5 weeks' gestation. In these six cases, the serum hCG level was extremely high. Serum screening results were available in seven cases in which preeclampsia did not develop, and the hCG levels were under 0.09 multiples of the median in five of the seven cases. In all six cases in which preeclampsia or hypertension developed, there was sonographic evidence of placentomegaly. Sonographic findings in 16 of 17 cases revealed fetal growth restriction, oligohydramnios, fetal anomalies, placentomegaly, or a combination of these. CONCLUSION: In our series of pregnancies complicated by triploidy, the risk of developing preeclampsia or hypertension in the second trimester was 35%. It appears that elevated serum hCG levels and placentomegaly are associated with a higher risk of preeclampsia but low hCG levels are not. This information is important in counseling patients who are hesitant to terminate a pregnancy purely for a fetal abnormality, even if the anomaly is lethal. PMID- 9397096 TI - Elevated serum nicked and urinary beta-core fragment hCG in preeclamptic pregnancies. AB - OBJECTIVE: To determine whether different molecular forms of hCG in serum and urine are elevated in preeclamptic pregnancies. METHODS: Forty-three pregnant women were studied: 25 preeclamptic women and 18 normotensive women. Immediately after blood and urine samples were collected, the protease inhibitors leupeptin (0.35 mM) and phenanthroline (22 mM) were added. Various molecular forms of hCG in serum (complete hCG, nonnicked hCG, complete free beta hCG) and in urine (complete hCG, beta-core fragment hCG) were measured by matched immunoassays with a common enzyme-labeled tracer antibody. The nicked hCG assay used a coating of beta-subunit monoclonal antibody with the addition of scavenger antibody to remove nonnicked hCG. Mann-Whitney U test and chi 2 test were used for statistical analyses. RESULTS: Preeclamptic women had significantly higher median (range) levels of serum complete and nicked hCG than did normotensive women (3620 [850-12,000] versus 2420 [310-4840] ng/mL, P = .024; and 102 [45-275] versus 71 [11-143] ng/mL, P = .010, respectively). Both median (range) urinary complete hCG creatinine and beta-core fragment-creatinine ratios were significantly higher in preeclamptic women than in normotensive women (37.6 [0.5-185] versus 11.3 [1.9 54], P = .013; and 11.8 [2-67] versus 5.3 [0.3-29], P = .009, respectively). CONCLUSIONS: Various molecular forms of hCG in serum and urine were significantly higher in preeclamptic than in normotensive pregnancies. PMID- 9397097 TI - Random protein-creatinine ratio for the quantitation of proteinuria in pregnancy. AB - OBJECTIVE: To compare random urine protein-creatinine ratios with 24-hour urine protein excretion rates in patients hospitalized with hypertensive disorders in pregnancy. METHODS: All hospitalized, hypertensive patients requiring 24-hour urine protein excretion collections were eligible for the study. During the 24 hour urine collection a separate 2-mL aliquot was taken for a protein and creatinine determination. RESULTS: Seventy-one samples were collected from patients with the following diagnoses: gestational hypertension (n = 56), preexisting hypertension and superimposed gestational hypertension (n = 11), and syndrome of hemolysis, elevated liver enzymes and low platelets (n = 4). The correlation coefficient between the random protein-creatinine ratio and the 24 hour urine protein excretion was 0.94. Calculated excretion rates with at least 300 mg protein in 24 hours had a sensitivity of 0.93, specificity of 0.90, and positive and negative predictive values of 0.87 and 0.95, respectively. For those samples with calculated excretion rates at least 5 g protein in 24 hours, the sensitivity was 1.00, specificity was 0.99, and positive and negative predictive values were 0.75 and 0.99, respectively. CONCLUSION: In nonambulatory hypertensive pregnant patients, there is a strong correlation between random voided protein-creatinine ratios and 24-hour urine protein excretions. PMID- 9397098 TI - A comparison between two doses of intravaginal misoprostol and gemeprost for induction of second-trimester abortion. AB - OBJECTIVE: To compare the abortifacient efficacies of two intravaginally administered misoprostol doses and gemeprost in termination of second-trimester pregnancy. METHODS: Eighty-one women between 12 and 24 weeks' gestation requesting abortion were randomized to receive intravaginally either 100 micrograms of misoprostol at 6-hour intervals (n = 27), 200 micrograms of misoprostol at 12-hour intervals (n = 26), or 1.0 mg of gemeprost at 3-hour intervals (n = 28). The regimen was continued until abortion, or for 36 hours, with assessment of the rate of complete and incomplete abortions as well as side effects within 48 hours from the start of the treatment. RESULTS: The final rates of terminations were 74% in the 100-microgram misoprostol group, 92% in the 200 microgram misoprostol group, and 89% in the gemeprost group. Abortion was complete in 37%, 61%, and 32% in each group, respectively (P = .03, when the 200 microgram misoprostol group was compared with the two other groups). The induction-to-abortion interval was longer (P = .001) in the misoprostol groups (mean 23.1 hours for the 100-microgram and 27.8 hours for the 200-microgram dose) than in the gemeprost group (14.5 hours). There was less pain (P = .01), diarrhea (P = .001), and vomiting (P = .01) in the misoprostol groups than in the gemeprost group. The mean blood loss in the misoprostol groups was lower than in the gemeprost group (P = .001). CONCLUSION: Intravaginal application of 200 micrograms of misoprostol at 12-hour intervals in induction of second-trimester abortion is equally effective to a standard gemeprost regimen. Misoprostol causes fewer side effects and is cheaper and more practical to use. PMID- 9397099 TI - Opportunities for prevention of perinatal group B streptococcal disease: a multistate surveillance analysis. The Neonatal Group B Streptococcal Disease Study Group. AB - OBJECTIVE: To evaluate the potential impact of ACOG and Centers for Disease Control and Prevention (CDC) consensus strategies for the prevention of perinatal group B streptococcal disease. METHODS: We evaluated cases of early-onset group B streptococcal disease identified by active surveillance during 1995, in four areas in North America with an aggregate 186,000 births per year. We reviewed the hospital records of mothers and infants and any prenatal records available on site. Cases were determined to be preventable based on whether group B streptococcal screening could have been performed prenatally, sensitivity of screening, presence of obstetric complications, and opportunity to administer antibiotics. RESULTS: We reviewed records for 245 of 246 infants with early-onset group B streptococcal disease in the surveillance areas. Most of the 53 case mothers who delivered preterm and 192 who delivered full-term had had at least one prenatal visit (83% and 99%, respectively). Few case-mothers had prenatal group B streptococcal screening cultures, although compliance was high for other prenatal screening tests. Fifty-four percent of case-mothers had a recognized obstetric risk factor for group B streptococcal disease: labor or rupture of membranes at less than 37 weeks, rupture of membranes for 18 hours on longer, or temperature 38C or greater. The estimated preventable portion of early-onset group B streptococcal cases was 78% for the screening-based approach (range 74% to 82% by area), compared with 41% for the risk-based approach (range 39% to 53% by area). CONCLUSION: Comprehensive implementation of either of the recommended prevention strategies could potentially prevent a substantial proportion of early onset group B streptococcal disease. PMID- 9397101 TI - Extemporaneous preparation of misoprostol gel for cervical ripening: a randomized trial. AB - OBJECTIVE: To compare the safety and efficacy of intravaginal misoprostol gel with that of tablets for ripening the cervix and inducing labor in women with unfavorable cervices. METHODS: Four hundred sixty-seven gravidas were randomized to receive misoprostol tablets (n = 234) or misoprostol gel (n = 233). The gel was prepared in the antepartum unit immediately before use by dissolving the tablet in 1 mL normal saline and mixing with 4 mL hydroxyethylcellulose gel. In both groups, a 50-microgram dose was applied intravaginally every 8 hours for two doses, then increased to 100-microgram for a total of six applications or 500 micrograms. RESULTS: The mean interval in hours from drug administration to start induction or labor (13.8 versus 18.2) and delivery (22.4 versus 29.0) was significantly less in the tablet group than in the gel group (P < .01 for both). Oxytocin and epidural use and the mean number of misoprostol insertions (1.4 versus 1.9) were lower in the tablet group than in the gel group (P < .05 for all). The incidences of tachysystole (13.7 versus 7.3%) and hyperstimulation (15.8 versus 7.7%) were significantly higher in the tablet group than in the gel group. Cesarean delivery rates and neonatal outcomes were similar between the groups. CONCLUSION: Intravaginal misoprostol gel is associated with fewer uterine contractile abnormalities than the tablet form of the drug but results in a slower time to labor or delivery. PMID- 9397100 TI - Vaginal birth after cesarean delivery: an admission scoring system. AB - OBJECTIVE: To develop a scoring system to predict the likelihood of vaginal birth in patients undergoing a trial of labor after previous cesarean delivery using factors known at the time of hospital admission. METHODS: Trial of labor was attempted in 5022 patients who were assigned randomly to score derivation and score testing groups. Multivariate logistic regression modeling was used in the score derivation group to develop a predictive scoring system for vaginal birth. The scoring system was then applied to the testing group to evaluate its predictive ability. RESULTS: Five variables significantly affected the mode of birth and were incorporated into a weighted scoring system. Rates of successful vaginal birth after cesarean ranged from 49% in patients scoring 0-2 to 95% in patients scoring 8-10. Increasing score was associated linearly with increasing probability of vaginal birth after cesarean. CONCLUSION: Increasing scores correlate with increasing probability of vaginal birth after cesarean. The admission vaginal birth after cesarean scoring system may be useful in counseling patients regarding the option of vaginal birth or repeat cesarean delivery. This information could be particularly valuable for the patient who opts for trial of labor but has second thoughts about her mode of birth when labor begins. PMID- 9397102 TI - The effect of pregnancy on cyclosporine levels in renal allograft patients. AB - OBJECTIVE: To assess the effects of pregnancy on cyclosporine levels in six renal allograft patients. METHODS: Maternal demographic, laboratory, clinical, and perinatal outcome data were recorded in six pregnant women with previous renal allografts receiving cyclosporine immunosuppression. The cyclosporine and serum creatinine levels were measured before pregnancy, during each trimester, and postpartum. RESULTS: The mean (standard deviation [SD]) maternal age was 29.1 (3.8) years. Parity ranged from 0 to 3. Mean serum creatinine levels tended to be lower during pregnancy than before or after, as did the mean cyclosporine levels. After adjusting for dose, five of six patients had declines in cyclosporine level during pregnancy. The mean (SD) gestational age at delivery was 37.5 (2.8) weeks with a mean (SD) birth weight of 2837 (538) g. CONCLUSION: Pregnancy in patients with renal allografts can lead to a substantial decline in cyclosporine levels. PMID- 9397103 TI - Maternal deaths due to homicide and other injuries in North Carolina: 1992-1994. AB - OBJECTIVE: To determine the role of homicide and other injuries in maternal deaths in North Carolina over the three-year period from 1992 through 1994. METHODS: Maternal deaths were identified from death certificates that indicated a maternal death and through an enhanced surveillance system that matches death certificates with live-birth and fetal-death certificates. Deaths were classified as direct, indirect, medically unrelated, or injury-related. Patterns of prenatal care were ascertained from the matching live-birth or fetal-death certificates. Maternal death rates for whites and nonwhites were calculated. RESULTS: The most common cause of maternal death was injury, accounting for 62 of the 167 deaths (37%). Homicide was the most common cause of injury-related death (35.5%). The relative risk of maternal death for nonwhites compared with whites was 1.8 (95% confidence interval [CI] 1.6, 2.1). Similarly, their relative risk for injury related maternal death was 1.7 (95% CI 1.4, 2.2). CONCLUSION: It is essential to include an analysis of injury-related deaths in maternal mortality reporting. As the most common cause of maternal deaths, injury is not limited to densely populated, metropolitan areas. Counseling regarding injury prevention, domestic violence, and depression should be a part of both prenatal and postpartum care. PMID- 9397104 TI - Internal and external anal sphincter anatomy as it relates to midline obstetric lacerations. AB - OBJECTIVE: To examine the anatomy of the internal and external anal sphincters in the area of midline obstetric lacerations, to gain insight into sphincter damage and repair. METHODS: The length, craniocaudal extent, and overlap of the internal and external anal sphincters in the perineal body were measured in 17 cadavers. Further anatomic observations were made in four sets of whole pelvis cross sections taken in the sagittal, coronal, and transverse planes. During the repair of 20 acute fourth-degree lacerations, observations were made to determine the internal sphincter visibility following birth. RESULTS: The external and internal and sphincters overlap by 17.0 mm (standard deviation [SD] 6.9), with the internal sphincter lying between the external sphincter and the anal canal. The internal sphincter extends an additional 12.2 mm (SD 5.9) cranial to the proximal extent of the external sphincter, whereas the caudal margin of the internal sphincter lies 3.7 mm (SD 7.2) cranial to the distal margin of the external sphincter. In pregnant women who sustained a fourth-degree laceration, we found that the internal sphincter can be identified as a rubbery white layer adjacent to the anal submucosa lying between the external sphincter and the anal canal. CONCLUSION: The internal anal sphincter lies between the anal mucosa and the external anal sphincter and extends more than a centimeter above the cranial margin of the external sphincter, a region where it is disrupted when a fourth degree obstetric laceration has occurred. PMID- 9397105 TI - Paracrine and intracellular signaling mechanisms of calcium waves in cultured human uterine myocytes. AB - OBJECTIVE: To establish mechanisms of intercellular communication in human myometrium other than action potential propagation. METHODS: Monolayer cultured human myometrium was used as a model system. The calcium-sensitive fluorescent dye, calcium green-1, was used as a probe for the concentration of intracellular free calcium. Intercellular calcium waves were initiated by mechanical stimulation and observed with video spectrofluorimetry. This technique allowed initiation of calcium waves from a known location at a known time while simultaneously controlling the flow rate of the bathing solution across the surface of the cells. Intercellular calcium waves were observed at bath flow rates between 0 and 5.1 mL/minute through a 0.4 mL chamber. Experiments were performed using low calcium, high potassium bathing solutions to eliminate the possibility of action potential signaling. RESULTS: In still bathing solution, calcium waves radiated symmetrically from the site of initiation. With the bathing solution flowing, two mechanisms of intercellular calcium wave propagation were observed-one dependent on and one independent of the direction of bath flow. The calcium waves that were independent of bath flow used an intracellular mechanism for intercellular communication, were only observed within 100 microns of the site of wave initiation, and demonstrated mean (+/- standard deviation [SD]) wave speeds of 14.1 +/- 2.6 microns/second. The waves dependent on bath flow used an extracellular signaling mechanism, were observed at distances much greater than 100 microns, and exhibited downstream biasing. Mean (+/- SD) wave speeds of flow-dependent calcium waves were faster under flow conditions than still bath conditions (36.0 +/- 4.7 versus 6.2 +/- 1.3 microns/second; P < .001). By exposing the cells to flurbiprofen, a water soluble prostaglandin synthetase inhibitor, both types of calcium waves were inhibited. CONCLUSION: Human myocytes demonstrate paracrine and intracellular signaling mechanisms for intercellular communication that are distinctly different from action potential propagation. PMID- 9397106 TI - Amniotic fluid glycine-valine ratio and neonatal morbidity in fetal growth restriction. AB - OBJECTIVE: To test the hypothesis that an elevated amniotic fluid glycine-valine ratio predicts neonatal morbidity in growth-restricted newborns. METHODS: Amniotic fluid (AF) was collected from 122 third-trimester pregnancies (range 31 39 weeks), 49 of which were complicated by fetal growth restriction. Amino acid analysis was performed by high-pressure liquid chromatography. Glycine-valine ratios were compared between normal and growth-restricted fetuses. Neonatal morbidity within the group of growth-restricted fetuses was characterized by evaluation of neonatal hypoglycemia, arterial cord blood gas analysis, and birth weight percentile. We also examined the correlation of AF glycine-valine ratio to the umbilical artery resistance index. The median interval between AF sampling and delivery was 1 day (range 0-8 days). Analyses were performed by Student t test, chi 2 with Yates correction, or simple correlation when appropriate. P < .05 was considered significant. RESULTS: Growth-restricted fetuses have a significantly elevated AF glycine-valine ratio compared with control subjects (3.31 +/- 1.06 versus 2.61 +/- 0.77, respectively, P < .001). There was no association of the glycine-valine ratio with gestational age for either group. An elevated glycine-valine ratio was not associated with neonatal hypoglycemia within the growth-restricted group (hypoglycemia: [n = 16] 3.19 +/- 1.07; no hypoglycemia: (n = 30) 3.44 +/- 1.09). There were no significant correlations of glycine-valine ratio with arterial cord blood pH (r = -0.10), oxygen pressure (r = 0.04), or base deficit (r = 0.12). There were no significant correlations of glycine-valine ratio and birth weight percentile (r = -.24) or umbilical artery resistance index (r = -.14). CONCLUSION: Amniotic fluid glycine-valine ratio is elevated in growth-restricted fetuses compared with control fetuses. However, the level of glycine-valine elevation is not associated with neonatal morbidity related to hypoglycemia, arterial cord blood gas abnormalities, or birth weight percentile. PMID- 9397107 TI - Significance of a false-positive trisomy 18 multiple-marker screening test. AB - OBJECTIVE: To determine if a false-positive trisomy 18 multiple-marker screening test (all three analytes low: maternal serum alpha-fetoprotein [AFP] at most 0.75 multiples of the median [MoM], unconjugated estriol at most 0.60 MoM, and hCG at most 0.55 MoM) indicates increased risk for obstetric complications or is related to maternal weight. METHODS: We accessed our genetic database to obtain multiple marker screening test results, fetal karyotypes, and pregnancy outcomes from all patients with a normal multiple-marker screening test (n = 3900) and from all patients with a positive trisomy 18 screening test (n = 103) seen in the prenatal diagnosis clinic from 1992 to 1996. During this period, only maternal serum AFP was adjusted for maternal weight. RESULTS: A positive trisomy 18 screen identified five of 12 trisomy 18 fetuses. Women with a false-positive trisomy 18 screen were heavier (175.6 +/- 43.8 lb versus 159.9 +/- 37.9 lb, P < .001) and younger (29.7 +/- 6.5 years versus 32.3 +/- 6.5 years, P < .001) than women with a normal multiple-marker screening test, but were not at increased risk for pregnancy complications. Weight-adjusting all three analytes reduced the false positive trisomy 18 screen rate by 42% (from 1.9% to 1.1%) but did not change the trisomy 18 detection rate. CONCLUSION: A false-positive trisomy 18 screening test does not indicate increased risk to develop pregnancy complications and may be related to inadequate correction for increased maternal weight. PMID- 9397108 TI - Fetal nuchal translucency thickness at 10-14 weeks' gestation and congenital diaphragmatic hernia. AB - OBJECTIVE: To examine the possible association between increased fetal nuchal translucency thickness at 10-14 weeks and congenital diaphragmatic hernia. METHODS: This was a multicenter ultrasound screening study for chromosomal defects in singleton pregnancies by a combination of maternal age and fetal nuchal translucency at 10-14 weeks' gestation. The prevalence of diaphragmatic hernia diagnosed prenatally or postnatally was calculated in the chromosomally normal group and in those pregnancies resulting in live births with no dysmorphic features suggestive of a chromosomal abnormality. We calculated the sensitivity of nuchal translucency above the 95th centile of the normal range in the detection of diaphragmatic hernia and the possible prognostic value of increased nuchal translucency in the prediction of outcome. RESULTS: There were 78,639 pregnancies presumed to be normal chromosomally, including 19 with diaphragmatic hernia. In four cases, the parents opted for termination of the pregnancy. The other 15 pregnancies resulted in live births; nine infants survived after successful surgical repair of the hernia, but six neonates died because of pulmonary hypoplasia. At the 10- to 14-week scan, the fetal nuchal translucency was above the 95th centile for crown-rump length in seven (37%) cases of diaphragmatic hernia. The translucency was increased in five of the six cases that resulted in neonatal death, compared with two of the nine survivors (Z = 2.32, P < .05). CONCLUSION: The prevalence of diaphragmatic hernia in chromosomally normal fetuses is about one in 4000, and nearly 40% of affected fetuses have increased nuchal translucency at 10-14 weeks' gestation. Increased nuchal translucency may be a marker of intrathoracic compression-related pulmonary hypoplasia. PMID- 9397109 TI - Cerebral and umbilical vascular resistance response to vibroacoustic stimulation in growth-restricted fetuses. AB - OBJECTIVE: To test the hypothesis that after vibroacoustic stimulation the ratio between cerebral vascular and umbilical vascular resistance in the growth restricted fetus is different from that in the normal fetus. METHODS: The pulsatility index (PI) of the middle cerebral artery and that of the umbilical artery (UA) were measured by pulsed Doppler velocimetry in 30 normal and 14 growth-restricted fetuses before and after vibroacoustic stimulation. The ratios of cerebral PI to UA PI and the changes in PI after vibroacoustic stimulation were calculated. Comparisons were made using the Wilcoxon rank-sum test or signed rank test. The statistical power of the study was 80%. RESULTS: Mean (+/- standard deviation) cerebral PI values before vibroacoustic stimulation (1.50 +/- 0.29) in normals and 1.29 +/- 0.26 in the fetal growth restriction [FGR] group) and UA PI values (1.00 +/- 0.18 in normals and 1.15 +/- 0.24 in the FGR group) were significantly different between groups (P < .04) and significantly decreased after vibroacoustic stimulation (P < .05). Although the cerebral to UA PI ratios (1.50 +/- 0.38 in normals and 1.13 +/- 0.33 in the FGR group) were significantly different between groups (P < .008), the values remained the same after vibroacoustic stimulation (P = .39 and .80, respectively). In all fetuses the fetal heart rate accelerated after vibroacoustic stimulation. CONCLUSION: Cerebral vascular resistance was lower and umbilical vascular resistance higher in the growth-restricted fetuses than in normals. The vascular resistance response after vibroacoustic stimulation in the growth-restricted fetus was not significantly different from the response of the normal fetus, suggesting preservation of regulation of resistance. PMID- 9397110 TI - Middle cerebral artery velocimetry: different clinical relevance depending on umbilical velocimetry. AB - OBJECTIVE: To evaluate the role of cerebral velocimetry as a predictor of perinatal outcome in high-risk pregnancies. METHODS: Middle cerebral artery pulsatility index was measured in 576 high-risk pregnancies undergoing umbilical velocimetry. The results of both tests were evaluated with respect to the birth of small for gestational age (SGA) infants and adverse perinatal outcome, defined as perinatal death, cesarean delivery for fetal distress, or low Apgar score. RESULTS: Once umbilical velocimetry was taken into account, cerebral velocimetry did not improve the prediction of fetal growth restriction or adverse perinatal outcome. Neither test was able to predict adverse perinatal outcome in normally grown fetuses. As for SGA fetuses with adverse perinatal outcome, the simultaneous assessment of both umbilical and cerebral velocimetry did not improve diagnostic accuracy (kappa index 0.37 versus 0.41 for umbilical velocimetry only). However, within the group of high-risk pregnancies with abnormal umbilical velocimetry, the risk of being SGA and having an adverse perinatal outcome was doubled (relative risk 2.1, 95% confidence interval 1.1, 4.3) if cerebral velocimetry also was abnormal. CONCLUSION: The routine use of cerebral velocimetry in high-risk pregnancies adds little information beyond that obtained from umbilical velocimetry; however, it is useful in predicting SGA infants with adverse perinatal outcome when umbilical velocimetry is abnormal. PMID- 9397111 TI - A prospective evaluation of fetal pericardial fluid in 506 second-trimester low risk pregnancies. AB - OBJECTIVE: To measure fetal pericardial fluid in low-risk second-trimester pregnancies and to evaluate outcome for those with measurements greater than 2 mm. METHODS: Five hundred and six women were referred for sonography between 16 and 25 weeks' gestation for common obstetric indications (dating, fetal survey, and placental location) unrelated to an increased risk of anomalies. All cases were evaluated with two-dimensional and M-mode real-time ultrasonography with the use of a mechanical sector transducer. The maximum distance of the fetal hypoechoic cardiac rim was recorded. We reviewed maternal and infant charts for those with measurements greater than 2 mm. RESULTS: Median (range) maternal age was 25 (15-42) years. Median gravidity and parity were two (1-14) and one (0-11), respectively. Median estimated gestational age was 20.4 (16.3-24.9) weeks. Fetal pericardial fluid was seen in 360 of 506 (71%) fetuses. Of these 360 fetuses, the mean distance (+/- 2 standard deviation) of the fetal hypoechoic cardiac rim was 1.20 mm +/- 0.91 mm (95% confidence interval 1.15, 1.25). Among the 506 cases, the maximum measurement was 3 mm. Ten of the 506 (2%) cases had measurements greater than 2 mm. None of these ten fetuses had a cardiac structural abnormality or arrhythmia, and perinatal outcome was unremarkable. CONCLUSION: During second trimester fetal ultrasonographic examination, visualization of pericardial fluid up to 2 mm in the fetus with current high-resolution technology is common and should not be regarded as pathologic. PMID- 9397112 TI - Anti-M isoimmunization: management and outcome at the Ohio State University from 1969 to 1995. AB - OBJECTIVE: To review the management strategies and outcome in gravidas with anti M isoimmunization over the past 26 years at The Ohio State University. METHODS: Data collected from 115 pregnancies found to have anti-M antibody at The Ohio State University from September 1969 through February 1996 were reviewed retrospectively. We analyzed indirect antiglobulin tests, amniotic fluid with spectrophotometric examination, direct antiglobulin tests, M antigen status, antepartum course, and perinatal outcome. RESULTS: Anti-M antibody was found in 90 women who had 115 pregnancies over 26 years. Among those with positive indirect antiglobulin tests, 104 pregnancies had titers at or below 1:4. Only one patient with an initial low titer experienced more than a three-fold increase to 1:64. Two women underwent a total of eight amniocenteses when titers were at or above 1:128. Forty-two (60%) of the 70 infants tested were positive for M antigen. Nine infants required phototherapy. Eight of these infants were delivered preterm. There was an increase in the number of women seen with anti-M antibody in pregnancy at our institution, with nearly 10% of all gravidas with a positive antibody screen having anti-M alloantibodies. There were no cases of hemolytic disease of the newborn, mild or severe. CONCLUSION: The prevalence of anti-M isoimmunization may be increasing. The incidence of severe hemolytic disease of the newborn due to anti-M is extremely low. We found no cases in our review of 115 pregnancies, although there have been several cases of severe hemolytic disease of the newborn reported. If anti-M is detected in pregnancy, the titer is low (no more than 1:4), and there is no history of prior pregnancy complications suggesting a hemolytic disease process, we recommend no further testing other than an indirect antiglobulin test at 28 weeks to look for the emergence of other alloantibodies. However, if the initial titer is elevated or there is a concerning obstetric history, serial titers should be performed and amniocenteses reserved for rising titers. PMID- 9397113 TI - Variation in the incidence of uterine leiomyoma among premenopausal women by age and race. AB - OBJECTIVE: To quantify the incidence of uterine leiomyoma confirmed by hysterectomy, ultrasound, or pelvic examination according to age and race among premenopausal women. METHODS: From September 1989 through May 1993, 95,061 premenopausal nurses age 25-44 with intact uteri and no history of uterine leiomyoma were followed to determine incidence rates of uterine leiomyoma. The self-reported diagnosis was confirmed in 93% of the medical records obtained for a sample of cases. Using pooled logistic regression, we estimated relative risks (RRs) of uterine leiomyoma according to race and examined whether adjustment for other potential risk factors could explain the variation in the race-specific rates. RESULTS: During 327,065 woman-years, 4181 new cases of uterine leiomyoma were reported. The incidence rates increased with age, and the age-standardized rates of ultrasound- or hysterectomy-confirmed diagnoses per 1000 woman-years were 8.9 among white women and 30.6 among black women. After further adjustment for marital status, body mass index, age at first birth, years since last birth, history of infertility, age at first oral contraceptive use, and current alcohol consumption, the rates among black women were significantly greater for diagnoses confirmed by ultrasound or hysterectomy (RR 3.25; 95% confidence interval [CI] 2.71, 3.88) and by hysterectomy (RR 1.82; 95% CI 1.17, 2.82) compared with rates among white women. We observed similar RRs when the cohort was restricted to participants who reported undergoing a screening physical examination within the 2 years before baseline. CONCLUSION: A higher prevalence of known risk factors did not explain the excess rate of uterine leiomyoma among premenopausal black women. PMID- 9397114 TI - The efficacy and complications of laparoscopic presacral neurectomy in pelvic pain. AB - OBJECTIVE: To evaluate the efficacy and complications of laparoscopic presacral neurectomy in pelvic pain. METHODS: We reviewed records of 655 patients receiving laparoscopic conservative surgery and laparoscopic presacral neurectomy for diagnoses including adenomyosis with dysmenorrhea (n = 55), moderate and severe endometriosis with dysmenorrhea (n = 127), minimal and mild endometriosis with dysmenorrhea (n = 208), primary dysmenorrhea (n = 99), and chronic pelvic pain with or without pathologic disease (n = 166). Pain relief was evaluated at least 12 months postoperatively. RESULTS: Pain relief was evaluated in 527 patients. Significant pain relief (no pain or mild pain requiring no medication) was found in 22 (52%) of 42 women with adenomyosis, in 75 (73%) of 103 with moderate to severe endometriosis with dysmenorrhea, in 123 (75%) of 164 with minimal to mild endometriosis with dysmenorrhea, in 64 (77%) of 83 with primary dysmenorrhea, and in 84 (62%) of 135 with chronic pelvic pain. There were four major complications (0.6%) that required further surgery, including injury of the right internal iliac artery (n = 1) and chylous ascites (n = 3). Three cases (0.5%) had laceration of the middle sacral vein controlled during laparoscopy. In addition, 485 (74%) of the 655 patients complained of constipation after laparoscopic presacral neurectomy, which was relieved easily by medication. CONCLUSION: Presacral neurectomy can be performed safely and efficiently by laparoscopy and is a valuable alternative treatment for pelvic pain. PMID- 9397115 TI - Duration of pregnancy after carbon dioxide laser conization of the cervix: influence of cone height. AB - OBJECTIVE: To determine if carbon dioxide laser conization of the cervix is a risk factor for preterm delivery in subsequent gestations and to evaluate whether there is any relationship between cone height and duration of pregnancy. METHODS: Patients of fertile age who had carbon dioxide laser conization were followed for reproductive events. Cases were matched one-to-one with controls for known risk factors for preterm delivery. Pregnancy duration, rate of preterm birth, and mode of delivery were studied. Parametric and nonparametric tests were used for statistical analysis. Logistic regression analysis and Cox proportional hazard modeling were used to investigate the relationship between cone height and subsequent preterm delivery. RESULTS: Sixty-four women with singleton pregnancies after carbon dioxide laser conization and 64 controls were included in the study. Overall, no difference was found in the rate of preterm delivery and duration of pregnancy. However, women with cone height of at least 10 mm had a higher rate of preterm delivery than either those with cone height less than 10 mm (five of 23 versus one of 41, P = .01) or the controls (five of 23 versus three of 64, P < .05). Cone height of at least 10 mm remained significant in predicting the occurrence of preterm delivery and the duration of pregnancy after adjusting for known risk factors (odds ratio 11.1, P < .05). CONCLUSION: Cone height of at least 10 mm is an independent risk factor for the duration of pregnancy and for the occurrence of preterm delivery in the subsequent gestation. PMID- 9397116 TI - Evaluation of parturition and other reproductive variables as risk factors for urinary incontinence in later life. AB - OBJECTIVE: To assess specific parturition and reproductive variables as potential risk factors for urinary incontinence in later life. METHODS: A mail survey was conducted with a random sample of 1922 women members of a large health maintenance organization. Multivariate analysis was used to estimate the independent association between parturition factors, hysterectomy, hormone use, and incontinence. RESULTS: Completed surveys were returned by 939 women (49%), 682 of whom reported at least one episode of incontinence in the past 12 months or ever having been treated for incontinence. On univariate analysis, women with incontinence were more likely to be white and heavier and to have had a hysterectomy before age 45, at least one live birth, a postdate (at least 42 weeks' gestation) birth, a labor lasting longer than 24 hours, and exposure to oxytocin. The risk of incontinence increased significantly with the number of exposures to oxytocin. In a multivariate model including age, there was a significant association between incontinence and white race (odds ratio [OR] 1.8, 95% confidence interval [CI] 1.2, 2.8), body mass (OR for fourth quartile 3.0, 95% CI 1.8, 5.0), estrogen replacement (OR 1.9, 95% CI 1.3, 2.8) and oxytocin (OR 1.9, 95% CI 1.0, 3.6). Parity was also associated with incontinence (P < .05). CONCLUSION: This study supports previous findings of a positive association between urinary incontinence and body mass, parity, and use of estrogen. In addition, we found a significant independent association between exposure to oxytocin during labor and incontinence in later life. PMID- 9397117 TI - A simplified protocol for pessary management. AB - OBJECTIVE: To evaluate a simplified protocol for pessary management. METHODS: Women with symptomatic pelvic organ prolapse who opted for pessaries were enrolled in a prospective simplified protocol for pessary management. After the initial pessary fitting, they were seen at 2 weeks for reexamination and thereafter at 3- to 6-month intervals. RESULTS: One hundred ten women (mean age 65 years) were enrolled, and 81 (74%) of them were fitted successfully with a pessary. Life-table analysis showed that 66% of those who used a pessary for more than 1 month were still users after 12 months and 53% were still users after 36 months. The severity of pelvic prolapse did not predict the likelihood of pessary failure except in cases of complete vaginal eversion. Patients complaining of stress incontinence were less likely to have a successful pessary fitting and more likely to opt for surgery. Current hormone use and substantial perineal support do not predict greater likelihood of pessary fitting success. No serious complications from using the pessary were observed in the study sample. CONCLUSION: Stringent guidelines calling for frequent pelvic examinations during pessary use can be relaxed safely. Pessaries can be offered as a safe long-term option for the management of pelvic prolapse. PMID- 9397118 TI - Effect of postmenopausal estrogen replacement on circulating androgens. AB - OBJECTIVE: To determine the effect of estrogen replacement therapy (ERT) on serum androgen levels in postmenopausal women. METHODS: We measured serum dehydroepiandrosterone (DHEA), DHEA-sulfate, testosterone, estradiol (E2), LH, FSH, and sex hormone binding globulin in 8:00 AM fasting serum samples from a previous randomized, blinded, placebo-controlled crossover study in which 28 postmenopausal women (27 naturally menopausal) were given 2 mg/day of oral micronized estradiol. The treatment arms were 12 weeks with a 6-week washout. RESULTS: Estrogen replacement therapy raised mean (+/- standard error of the mean [SEM]) serum E2 from 8.7 +/- 1.0 to 117 +/- 18.7 pg/mL (P < .001 from baseline). Concurrently, mean (+/- SEM) DHEA-sulfate fell from 67.3 +/- 9.6 to 52.1 +/- 6.4 micrograms/dL (P < .001), and mean (+/- SEM) testosterone fell from 16.1 +/- 2.4 to 9.4 +/- 1.4 ng/dL (P = .006). Both FSH and LH declined significantly. Sex hormone binding globulin increased by 160% with ERT (P < .001). CONCLUSION: Menopausal ERT decreases serum androgen levels, decreasing DHEA-sulfate and testosterone by 23% and 42%, respectively. Whereas the decline in testosterone is likely due to decreased LH-driven ovarian stromal steroidogenesis, the declining levels of DHEA-sulfate also may imply a direct adrenal effect of estrogen. Bioavailable testosterone likely is reduced even more profoundly because sex hormone binding globulin is increased 160% by estrogen. Thus, menopausal ERT may induce relative ovarian and adrenal androgen deficiency, creating a rationale for concurrent physiologic androgen replacement. PMID- 9397120 TI - The 4-S modification of the Roeder knot: how to tie it. AB - BACKGROUND: The 4-S modification of the Roeder knot may be tied laparoscopically as a single-throw knot. TECHNIQUE: It is tied by adding a fourth wrap around the suture loop and securing the loop in place with a square knot rather than a single half-hitch. EXPERIENCE: We have used this knot in laparoscopic surgeries for more than 2 years and have not observed knot slippage. CONCLUSION: This modification results in a knot comparable in strength to the strongest laparoscopic multiple-throw square knots. PMID- 9397119 TI - Papanicolaou smears by the Bethesda system in endometrial malignancy: utility and prognostic importance. AB - OBJECTIVE: To evaluate the prognostic significance of the Bethesda system's cytologic categories in patients with endometrial malignancy. METHODS: Patients with biopsy or hysterectomy-proven endometrial malignancy and a Papanicolaou smear result reported using the Bethesda system within 1 year of diagnosis were identified through retrospective review of our computerized database. RESULTS: After introduction of the Bethesda system in our laboratory on November 1, 1992, until January 1, 1997, 112 eligible patients were identified (108 with carcinomas and four with carcinosarcomas). Patients with cytologic diagnoses of malignancy (n = 17) were significantly more likely to have International Federation of Gynecology and Obstetrics (FIGO) grade 3 tumors and high-risk histology (serous, clear cell, and adenosquamous carcinoma and carcinosarcoma) than those with atypical glandular cells of uncertain significance (n = 33) or those with cytology not suspicious for malignancy (n = 63). Patients with malignant smears were also significantly more likely to have cervical extension, malignant peritoneal cytology, and FIGO stage II, III, or IV than those with atypical glandular cells of uncertain significance or those with cytology not suspicious for malignancy. CONCLUSION: Papanicolaou smears obtained within 1 year of histologic diagnosis of endometrial malignancy and interpreted using the Bethesda system were suspicious for (atypical glandular cells of uncertain significance) or diagnostic of malignancy in nearly half of all cases (29 and 15%, respectively). Patients having malignant glandular cells were more likely to have poor prognostic pathologic findings. PMID- 9397121 TI - Three-dimensional method for determination of amniotic fluid volume in intrauterine pockets. AB - BACKGROUND: Although current ultrasound techniques provide a linear (amniotic fluid index; AFI) or two-dimensional area index of amniotic fluid (AF), these indices have limited correlation with actual AF volume. We sought to quantify the three-dimensional volume of ultrasound-identified AF pockets, as assessed by the AFI and two-dimensional area methods. The BVI 2500 (Bladder Volume Instrument 2500; Diagnostic Ultrasound Corp., Redmond, WA) has been used to quantify the volume of residual urine in the bladder. INSTRUMENT AND METHOD: The BVI 2500 (Diagnostic Ultrasound Corp.) ultrasound uses a rotating 2-MHz transducer, computer-defined fluid interface, and computer integration of 12 cross-sectional images to calculate three-dimensional fluid volume. After providing written informed consent, 14 term pregnant patients (36-42 weeks) were evaluated using the BVI 2500 and an Ultramark 8 sector scan (Advanced Technology Laboratory, Bothell, WA). The largest vertical fluid pocket in each quadrant of the abdomen was identified with the sector scan, and vertical and horizontal measurements for AFI and two-dimensional area were recorded. Simultaneous AF volume measurements of each pocket were performed three times with the bladder volume instrument, and maximum values were used. Three-dimensional volume, two-dimensional area, and AFI values were compared by correlation analysis, with P < or = .05 considered statistically significant. EXPERIENCE: Among all patients, the average (+/- standard deviation) AFI was 7.6 +/- 4.1 (range 1.5-16.4) cm, and the average two dimensional area was 30.9 +/- 21.1 (range 4.3-81.3) cm2. This corresponded to an average three-dimensional volume of 215 +/- 134 (range 23-497) cm3. Three dimensional volume correlated highly with both AFI (r = 0.9; P < .001) and two dimensional area (r = 0.86; P < .001). One AFI centimeter was equivalent to a volume of 30 cm3. CONCLUSION: There are highly significant linear correlations of three-dimensional amniotic fluid volumes with AFI and two-dimensional area. The four pockets used in AFI determination account for only 50% of total AF volume. Three-dimensional determinations may aid in clinical assessments of AF volume. PMID- 9397122 TI - A model clinical faculty workshop program for a multisite clerkship. AB - Over the past 20 years, the University of Washington Department of Obstetrics and Gynecology has conducted twice-yearly faculty development workshops at the University for clinical faculty at community sites spread over three time zones in Washington, Alaska, Montana, and Idaho. These workshops consist of three separate parts: 1) a session to report on student clerkship performance and faculty ratings, 2) a session on curriculum and/or teaching, and 3) a session to update medical topics. Faculty found the workshops useful for improving their clinical teaching, keeping in touch with the full-time faculty, and developing a sense of cohesiveness. The department found the workshops useful in maintaining the teaching of consistent content and ensuring that all sites provide students with comparable experiences and evaluation. PMID- 9397123 TI - Telomerase in gynecologic cancers. AB - OBJECTIVE: To provide the obstetrician-gynecologist with the following: 1) basic concepts of telomere shortening and telomerase activation and their relation to cellular immortalization and cancer, 2) an overview of potential uses of telomerase activity in cancer diagnosis, assessment of prognosis, and the development of new anticancer therapeutic approaches, and 3) a review of the literature on telomerase activity in gynecologic cancers. DATA SOURCES: A computerized search for articles published in which telomerase or telomerases were included as a subject heading or a textword was performed using the Ovid Search Software (Ovid Technologies Inc., New York, NY). The search was conducted of the following databases of the National Library of Medicine and the National Cancer Institute: MEDLINE January 1966 to May 1997, HealthSTAR January 1975 to May 1997, AIDSline January 1980 to May 1997, and CancerLit January 1980 to May 1997. Additional sources were identified through cross-referencing. METHODS OF STUDY SELECTION: All sources identified were reviewed with particular attention to human application, specifically in gynecologic cancers. TABULATION, INTEGRATION, AND RESULTS: A total of 304 references was identified. Each reference was reviewed to determine the relevant contribution to the basic understanding of the role of telomerase in cellular immortalization and the development of cancer, potential uses of telomerase measurement in cancer, gynecologic applications, and potential use of telomerase inhibitors in cancer therapy. CONCLUSION: Telomerase activity might be a valuable diagnostic and prognostic tool in gynecologic and other types of cancer, and telomerase inhibition might prove to be a significant therapeutic approach for some types of cancer. Better understanding of the relation between telomerase activation, tumor suppressor genes, and oncogenes might clarify several aspects of early tumorigenesis and result in development of novel approaches to early cancer detection and prevention. PMID- 9397124 TI - Obstetrics & Gynecology in 1996: marking the progress toward evidence-based medicine by classifying studies based on methodology. AB - OBJECTIVE: To test the hypothesis that researchers in obstetrics and gynecology favor an observational type of study design. DATA SOURCES: The 12 regular issues of Obstetrics & Gynecology published during 1996 were analyzed. METHOD OF STUDY SELECTION: All articles in the journal Obstetrics & Gynecology were reviewed for the year 1996, except that separate issues covering case reports, case condensations, and reviews were excluded. TABULATION, INTEGRATION, AND RESULTS: Studies were classified as observational or experimental. Observational studies were subclassified as either descriptive, case-control, or cohort. Experimental studies were subclassified as either randomized controlled trial (RCT) or uncontrolled trial. Other study designs were noted. Of the 316 studies published during 1996, 241 (76%) were observational, 43 (14%) experimental, and 32 (10%) other. There were 162 (51%) descriptive studies, 44 (14%) case-control studies, 35 (11%) cohort studies, 35 (11%) RCTs, and eight (3%) uncontrolled trials. CONCLUSION: Researchers who publish in Obstetrics & Gynecology favor an observational study design. With evidence-based medicine growing in popularity as a new standard or paradigm, the reliance on observational studies may have implications. PMID- 9397125 TI - Nifedipine and ritodrine in the management of preterm labor: a randomized multicenter trial. PMID- 9397126 TI - Mullerian agenesis: an update. PMID- 9397127 TI - Stress-point intervention for parents of repeatedly hospitalized children with chronic conditions. AB - Little is known about how to assist children with chronic conditions and their families cope with repeated hospitalizations. A two-group, pretest-posttest study was done to determine whether a community-based, stress-point nursing intervention for parents could decrease distress and improve child and family functioning. Fifty participants were randomly assigned to intervention or usual care control groups. The intervention focused on specific, parent-verified child and family issues. Three months after hospitalization, intervention parents had better coping and family functioning than those in the usual care group. Intervention parents' anxiety was initially higher and then lower. There were no child behavior differences between the groups after hospitalization. Intervention children had no developmental regression at 2 weeks and better developmental gains 3 months after discharge than the usual care children. Stress-point intervention for families and their children with chronic conditions improved family coping and functioning, and eliminated hospitalization-induced developmental regression. PMID- 9397128 TI - Attention, coping, and activity in children undergoing orthopaedic surgery. AB - The purpose of this study was to determine how children's preoperative focus of attention on the stresses of surgery related to their preoperative coping and return to usual activities during recovery. Children's attention was classified according to three different foci: concrete-objective, emotion, and vague. Children (N = 97) between the ages of 8 and 17 years who were undergoing major orthopaedic surgery participated in the study. Data were collected the day before surgery, and at 3, 6, and 9 months postoperatively. Children who focused on the concrete-objective aspects of surgery had the most positive activity outcomes, followed by the emotion-focused attention group. Children who were classified as having vague focused attentions had the least favorable activity outcomes. When there were significant coping by attention interaction effects, vigilant copers who had a concrete-objective focus of attention had the most favorable activity outcomes at each time of measurement. Children who were able to focus their attention on concrete aspects of the experience tended to use vigilant coping and were able to return to their usual activities sooner. PMID- 9397129 TI - Living with fibromyalgia: sense of coherence, perception of well-being, and stress in daily life. AB - Fibromyalgia (FM) is a chronic pain syndrome that has a considerable impact on the ill person's daily life. The purpose of this study was to describe levels of sense of coherence (SOC), perceptions of well-being, and stress in daily life in women with FM in comparison with healthy women, and to determine whether SOC is related to perceived levels of stress and well-being. Thirty women with FM were compared with 30 healthy women matched for Type A behavior. The results revealed a complex picture of the women with FM. On the one hand, they reported many symptoms but, on the other, they rated themselves as feeling quite well and experiencing an SOC in life, despite severe problems. The FM women with a stronger SOC perceived greater well-being than those with a weaker SOC. They felt more hopeful, more free, more valuable, and more like others. Results suggest that women with a weaker SOC may need extra support. More research is needed to investigate the experience of living with FM in order to discover what it is that makes life worthwhile despite high symptom levels. PMID- 9397130 TI - Nicotine dependence and smoking topography among black and white women. AB - Nicotine dependence is a complex phenomenon involving behavioral, biological, and pharmacological components that influence smoking cessation rates. The purpose of this study was to characterize the multidimensional aspects of nicotine dependence and cigarette smoking topography behaviors among Black and White women smokers. Thirty-seven women participated in a 2-hr protocol in the General Clinical Research Center. Plasma cotinine to cigarette ratio was significantly associated with three topography variables: total puff duration, total cigarette time, and carbon monoxide (CO) boost. Black women scored higher on plasma cotinine levels, cotinine per cigarette ratio, and CO increase pre- to postcigarette than White women. Implications for clinical practice include assessing nicotine dependence beyond self-reported cigarettes per day to develop more appropriate smoking cessation interventions. PMID- 9397131 TI - A causal functional explanation of maintaining a dependent elder in the community. AB - A causal functional structure demonstrates how a perpetual process, like caregiving, maintains itself. The caregiver functional learning model was used to identify factors influencing caregivers when maintaining dependent elders in the community. These factors include seriousness of illness, power, burden, care quality, self-assurance, an understanding of the elder's needs, and time spent in caregiving. This five-stage theoretical model was tested using a sample of 70 caregivers. Findings supported two of the theoretical relationships: seriousness of illness (beta = .60) with burden (R2 = 0.35), and self-assurance (beta = .36) with time (R2 = 0.18). Three new relationships were added with the empirical model: power (beta = .48) with burden (R2 = 0.52), seriousness of illness (beta = .36) with self-assurance (R2 = 0.11), and burden (beta = .33) with time (R2 = 0.24). A causal functional structure can be used to explain how the phenomenon of caregiving maintains itself. PMID- 9397132 TI - Family involvement in the nursing home: family-oriented practices and staff family relationships. AB - Staff-family cooperation in caring for elders in nursing homes is recommended but poorly understood. Family involvement and staff-family interactions in nursing homes with differing family orientations were investigated. Friedemann's (1995) system-based family theory guided the study. Of all 208 licensed nursing homes in southern Michigan, 143 completed a survey about their family-oriented practices. Family orientation was ranked accordingly. Twenty-four nursing homes were randomly selected to conduct semistructured telephone interviews with 177 family members. Data were analyzed by thematic interpretation. Findings showed a wide range of involvement patterns that promoted family connectedness, maintenance of control, growth, and learning. Families desired various types of staff cooperation and were given such opportunities in homes with high family orientation. PMID- 9397134 TI - Endings, secrets, and silences: overreading in narrative inquiry. AB - Qualitative researchers seek to understand the words of the people they interview and may use narrative inquiry to find meaning in the stories told by research participants. Narrative inquiry entails "overreading," a sensitivity to unspoken or indirect statement, which is central to interpretation. Some of the tools of the literary overreader are applied to two research interviews, particularly as they direct readers to attend to inconsistencies, endings, repetitions, and silence. Because overreading is also intrusive, we conclude with some considerations about how far the overreader of the research may legitimately go. PMID- 9397133 TI - Determination of reliability and validity in home monitoring data of pulmonary function tests following lung transplantation. AB - Electronic spirometry units were used to monitor lung transplantation recipients upon their return home. The data from 77 participants were used to develop methods to verify that the pulmonary function measurements, forced vital capacity (FVC) and forced expiratory volume in 1 s (FEV1), were reliable and valid. The standard deviation was calculated for the best daily effort on consecutive days of home spirometry. An acceptable upper limit for the standard deviation, as the measure of day-to-day reliability, was 0.20 for FVC and 0.15 for FEV1. Validity was determined by examining the mean difference (bias) between the spirometry done in the pulmonary function laboratory and the home monitoring results. The clinic values were slightly higher, with an average difference of 0.15 for FVC and 0.12 for FEV1. Therefore, the home spirometry measurements have a high degree of reliability and validity and can now be used for early detection of serious complications. PMID- 9397135 TI - Excretion of epidermal growth factor in human adult polycystic kidney disease. AB - In chronic renal failure, epidermal growth factor (EGF) excretion is decreased. In this study, asymptomatic adult polycystic kidney disease (APKD) patients with a relatively preserved glomerular filtration rate were examined. Excretion of EGF was studied in 6 patients with APKD (median age 42 years; serum creatinine [median] 95 [range-80-133] mumol/l) and compared with that of 28 healthy controls. EGF was determined in a spot morning urine by using a specific radioimmunoassay, and expressed in relation to creatinine excretion. Excretion of EGF in APKD was (median) 157 (range-13-359) and in the controls (median) 546 (range-238-1199) pmol/mmol creatinine (p < 0.001). Low excretion of EGF in APKD patients with preserved kidney function suggests a distal abnormality at an early stage of the disease, prior to the development of renal failure. PMID- 9397136 TI - The Israeli Diagnostic Center for Malignant Hyperthermia: 7-years' accumulated experience. AB - Malignant hyperthermia (MH), a rare pharmacogenetic trait, can be lethal when susceptible individuals are exposed to triggering agents during general anesthesia. We present our experience with the caffeine-halothane in vitro contracture test (IVCT) for the diagnosis of malignant hyperthermia susceptibility. Out of 75 patients that were referred for consultation to the MH diagnostic center over a period of 7 years, we performed muscle biopsies and IVCT in 21 patients. A total of 6 patients were found to be MH-positive. Appropriate recommendations for future anesthetic management, and additionally, for testing the immediate family were made following a positive diagnosis. Improved familiarity with the syndrome of MH, and performance of IVCT when family or clinical history suggest malignant hyperthermia susceptibility, are imperative measures to prevent the potential fatality associated with this syndrome. PMID- 9397137 TI - Flow parameters of the normal arterial duct in the fetus. AB - The increasing interest in arterial duct flow patterns in the fetus warrants the establishment of an accurate range of normal flow parameters throughout gestation. We therefore undertook a prospective echocardiographic study of 181 normal fetuses from the 16th to the 40th week of gestation. Adequate Doppler interrogation of the duct was obtained in 71% of the fetuses examined. Peak gradient, mean gradient and flow velocity integral in systole and diastole were digitized. The peak systolic gradient throughout pregnancy measured 2.7 +/- 1.4 mmHg with a slight tendency to increase with gestational age (r = 0.58). The peak ratio, defined as peak systolic gradient divided by peak diastolic gradient (28.1 +/- 14.9), did not vary significantly with gestational age. This time-independent index complements peak systolic flow in the assessment of normal and abnormal ductal flow. The definition of the normal range for ductal flow parameters, based on this relatively large fetal population, should facilitate the accurate diagnosis of fetal duct constriction. PMID- 9397138 TI - Alveolar macrophages fat stain in early diagnosis of fat embolism syndrome. AB - The aim of this study was to assess the contribution of bronchoalveolar lavage (BAL) in the diagnosis of fat embolism syndrome (FES). The presence of fat droplets in alveolar macrophages was addressed in 13 trauma patients with bone fractures and 10 non-trauma patients with acute respiratory distress syndrome (ARDS). The control group was composed of 5 anesthesized patients with ischemic heart disease, immediately prior to cardiac surgery. Two patients with suggestive clinical and laboratory signs of FES had 40% and 24% fat-containing alveolar cells, respectively. The trauma patients without signs of FES displayed a wide variation in the percentage of fat-containing macrophages (from 3% to 95%). Most of the patients with ARDS who were receiving lipid emulsion as part of their parenteral nutrition, had a high percentage (> 85%) of fat-containing macrophages. Patients with normal lungs had no fat-containing macrophages. Our findings suggest that BAL Oil Red O-positive macrophages are frequently observed in trauma patients irrespective of the presence of FES. Therefore, estimation of the percentage of fat-containing macrophages from BAL is an unreliable marker of FES. PMID- 9397139 TI - Pediatric cholesterol screening: missed opportunities. AB - The current recommendations for childhood cholesterol screening include screening children in whom 1) a parent/grandparent has premature heart or vascular disease or died suddenly; 2) a parent has an abnormal lipid profile; 3) the family history is unobtainable. Over a 3-year period, 256 children referred for hypercholesterolemia were evaluated for heritable hyperlipidemia. We reviewed their family histories and obtained lipoprotein profiles of all of their immediate family members. Of these families, 89 parents had unsuspected hypercholesterolemia of whom 38, whose average age was 36 years, died of a myocardial infarction. In addition, 83 children with no family history of premature coronary artery disease or hypercholesterolemia, were diagnosed with inherited hyperlipidemia (25 with hetrozygous familial hypercholesterolemia, and 58 with familial combined hyperlipidemia). Thus, many adults have no awareness of hyperlipidemia prior to a fatal heart attack, nor of their children as having hyperlipidemia, and a large percentage of children with inherited hyperlipidemia would not have been diagnosed if all of their immediate family members (parents and siblings) had not been screened for a complete lipid profile. These results suggest that in addition to screening, all family members of hypercholesterolemic children, pediatricians and family practitioners should urge parents who may be unaware of their cholesterol levels or have no knowledge of their family history to undergo cholesterol screening in order to comply with NCEP guidelines calling for serum cholesterol measurements in all adults above the age of twenty. PMID- 9397140 TI - Speech disorders in Israeli Arab children. AB - The aim of this work was to study the frequency of speech disorders in Israeli Arab children and its association with parental consanguinity. A questionnaire was sent to the parents of 1,495 Arab children attending kindergarten and the first two grades of the seven primary schools in the town of Taibe. Eight-six percent (1,282 parents) responded. The answers to the questionnaire revealed that 25% of the children reportedly had a speech and language disorder. Of the children identified by their parents as having a speech disorder, 44 were selected randomly for examination by a speech specialist. The disorders noted in this subgroup included errors in articulation (48.0%), poor language (18%), poor voice quality (15.9%); stuttering (13.6%), and other problems (4.5%). Rates of affected children of consanguineous and non-consanguineous marriages were 31% and 22.4%, respectively (p < 0.01). We conclude that speech disorders are an important problem among Israeli Arab schoolchildren. More comprehensive programs are needed to facilitate diagnosis and treatment. PMID- 9397141 TI - A community hospital experience with colonoscopic polypectomies. AB - This study analyzed 432 consecutive polypectomies performed in 279 patients in the gastroenterology unit of a community hospital. The patients were separated into 2 groups; group I--symptomatic patients considered suitable for colonoscopic examination, and group II--asymptomatic high-risk patients. The mean number of detected polyps was similar in both groups, the vast majority of the polyps in both groups were small (< 5 mm), and were mainly of tubular histology. Polyps in the rectosigmoid area were more common (56.6%) in the symptomatic patients than in the asymptomatic patients (44.1%). Fourteen percent of patients in group I and 33% in group II had no polyps within 60 cm from the anal verge. Carcinoma in situ was found in large polyps mainly in group I. Flat adenomas were not found in the studied population. The incidence of hyperplastic polyps was similar in both groups and did not predict the concomitant existence of adenomatous polyps. The male:female ratio was the same in both groups. The percent of detected polyps increased with age. A strong right shift in the location of the polyps was evident with increasing age. Multiple polyps were a common event in this Israeli population of symptomatic and high-risk asymptomatic patients. More than 30% of the polyps were found outside the reach of the sigmoidoscope, with this proportion increasing with age. These data provide further support to the claim that colonoscopy should therefore serve as the choice diagnostic tool in these high-risk populations. PMID- 9397142 TI - Squamous cell carcinoma of the cervix with psoas abscess-like metastasis in an HIV-negative patient. AB - A psoas abscess-like metastasis of squamous cell carcinoma of uterine cervix was diagnosed in a 50-year-old, HIV-negative woman. While there was complete regression of the primary tumor, the psoas cystic lesion grew in size during chemotherapy, with iliac bone destruction and invasion to the gluteus muscle. A computed tomography-guided bone biopsy was done and histopathologic examination revealed squamous cell carcinoma consistent with the primary lesion. Intratumoral injection of cisplatin and radiotherapy resulted in partial relief of symptoms. This form of aggressive, bone destruction metastasis was hitherto reported only in association with AIDS. PMID- 9397143 TI - Littoral cell angioma: a vascular tumor mimicking a solid tumor on a Tc-99m-red blood cell spleen scan. AB - A splenic space occupying lesion, in a 45-year-old woman, was negative in a Tc 99m-RBC spleen scan. A diagnostic splenectomy was performed and the lesion was found to be a vascular tumor, lately identified as littoral cell angioma. The histological and immunohistochemical findings are discussed in correlation with the imaging results. PMID- 9397144 TI - Primary biliary cirrhosis associated with antiphospholipid syndrome. AB - A 47-year-old female was admitted for severe pain of 1 month's duration in the third and fourth toes of the right foot, culminating in gangrene. Laboratory findings revealed liver enzyme abnormalities, and anti-mitochondrial, anti phospholipid and antinuclear and doubtful anti-DNA antibodies. Systemic lupus erythematosus (SLE) was excluded on clinical grounds after a 6-year follow-up. Therefore, a diagnosis was made of the primary antiphospholipid syndrome, complicated by microvasculopathy, and associated with primary biliary cirrhosis. PMID- 9397145 TI - Bilateral brachial plexopathy after E. coli sepsis. AB - A 67-year old patient, operated for closure of ileostomy 3 days before, developed E. coli sepsis with suspected peritonitis. Two days later, pain and weakness in the shoulder girdle, scapula and proximal upper limb muscle appeared simultaneously, followed by marked atrophy. Electromyography (EMG) examination manifested bilateral active denervation of upper brachial plexus. Any trial to isolate a pathogenic factor other than the E. coli failed. Due to the bilateral proximal upper limb's paralysis, grooming and upper-body dressing obliged the patient to ask for complete assistance. In other daily living activities he was partially independent. PMID- 9397147 TI - Gastrointestinal events related to the use of non-steroidal anti-inflammatory drugs: some clinical and economic aspects. PMID- 9397146 TI - Glucagon-like peptide-1 structure, function and potential use for NIDDM. AB - Basic research on the cellular mechanisms that control the expression of the gene encoding glucagon has led to the discovery of proglucagon. This precursor is processed by tissue-specific proteolysis to produce glucagon in pancreatic alpha cells and a glucagon-like peptide-1 (GLP-1) in the intestine. GLP-1 is a hormone that is released by intestinal cells into the circulation in response to food intake. GLP-1 and gastric inhibitory peptide (GIP) which has also been termed glucose-dependent insulinotropic peptide appear to account for most of the incretin effect in the augmentation of glucose-stimulated insulin secretion. These two hormones have specific beta-cell receptors that are coupled to GTP binding proteins to induce production of cyclic AMP and activation of cyclic AMP dependent protein kinase. It is proposed that at least one factor contributing to the pathogenesis of non-insulin-dependent diabetes mellitus (NIDDM) is desensitization of the GLP-1 receptor on beta-cells. At pharmacological doses, infusion of GLP-1, but not of GLP, can improve and enhance postprandial insulin response in NIDDM patients. Agonists of GLP-1 receptor have been proposed as new potential therapeutic agents in NIDDM patients. The observations that GLP-1 induces both secretion and production of insulin, and that its activities are mainly glucose-dependent, led to the suggestion that GLP-1 may present a unique advantage over sulfonylurea drugs in the treatment of NIDDM. PMID- 9397148 TI - Obesity epidemic puts millions at risk from related diseases. PMID- 9397149 TI - Polio eradication effort marches on despite wars. PMID- 9397150 TI - All-trans retinoic acid blocks the antiproliferative prodifferentiating actions of 1,25-dihydroxyvitamin D3 in normal human keratinocytes. AB - 1,25-Dihydroxyvitamin D3 [1,25(OH)2D3] and all-trans retinoic acid (RA), the active metabolites of vitamins D and A respectively, regulate the proliferation and differentiation of keratinocytes. Both the vitamin D receptor (VDR) and the retinoic acid receptor family (RAR) bind to DNA response elements as heterodimers with the retinoic X receptor (RXR), suggesting that there are pathways of action that are shared by both compounds. Therefore, we examined the interactions of 1,25(OH)2D3 and RA upon the proliferation and differentiation of normal human keratinocytes (NHK) and of a squamous cell carcinoma cell line, SCC4. Although both 1,25(OH)2D3 and RA were each able to inhibit NHK proliferation in a dose dependent manner, when they were administered in combination, proliferation was stimulated, suggesting mutual antagonism. In contrast, SCC4 cells proved insensitive in terms of proliferation to 1,25(OH)2D3 and to all but the highest concentration (10(-6) M) of RA. 1,25(OH)2D3 exerted a biphasic effect on transglutaminase (TGase) and involucrin (INV) mRNA levels, with maximal stimulation at 10(-9) M. RA inhibited TGase and INV mRNA levels and antagonized the stimulation by 1,25(OH)2D3. A similar pattern was observed for TGase protein, but, RA, which, by itself, reduced INV, markedly enhanced the ability of 1,25(OH)2D3 to raise INV levels, possibly by inhibiting 1,25(OH)2D3-stimulated TGase activity and cross-linking of soluble INV into the insoluble cornified envelope (CE). Thus, in NHK cells, RA antagonizes the antiproliferative prodifferentiating actions of 1,25(OH)2D3, but assessment of a single marker, such as INV protein, may be misleading. PMID- 9397151 TI - Epidermal growth factor (EGF) receptor density controls mitogenic activation of normal rat kidney (NRK) cells by EGF. AB - Normal rat kidney (NRK) fibroblasts are immortalized cells that are strictly dependent on externally added growth factors for proliferation. When cultured in the presence of epidermal growth factor (EGF) as the only growth stimulating hormone, these cells have a normal phenotype and undergo density-dependent growth inhibition. It has been postulated that this density-arrest results from a decrease of EGF receptor levels below a threshold level which makes these cells unresponsive to stimulation by EGF. In the present study, we show that NRK cells, made quiescent by serum-deprivation at submaximum density, are mitogenically still responsive to EGF, but show enhanced mitogenic stimulation after 8 hr pre treatment with either transforming growth factor beta (TGF beta) or retinoic acid (RA), while prostaglandin F2 alpha (PGF2 alpha) and bradykinin (BK) enhance the mitogenic stimulation by EGF only slightly under these conditions. Addition of TGF beta or RA results in an increase of both 125I-EGF-binding capacity and EGF receptor mRNA levels. Using flow cytometric analysis, we show that pre-treatment with TGF beta or RA increases the percentage of cells entering the cell cycle as a function of time. Furthermore, pre-treatment of the cells with TGF beta or RA increases the rate of mitogen-activated protein kinase (MAPK) phosphorylation by EGF. PGF2 alpha and BK also increase EGF receptor levels, but only with delayed kinetics. These results show that already in serum-deprived quiescent NRK cells, EGF receptor levels limit EGF-induced mitogenic stimulation. This observation provides further evidence for the regulating role of the EGF receptor in density dependent growth control of NRK cells. PMID- 9397152 TI - Analysis of the transcriptional activity of the 5'-flanking region of the rat catalase gene in transiently transfected cells and in transgenic mice. AB - Transiently transfected cell lines and transgenic mice were used to study the transcriptional activity of the 5'-flanking region of the catalase gene. Fragments of the 5'-flanking region of the rat catalase gene ranging in length from 3,421 base pairs (bp) to 69 bp were fused to the chloramphenicol acetyltransferase (CAT) reporter gene, and the transcriptional activity of the reporter gene was measured following transient transfection in three cell lines: a human hepatoma cell line (HepG2), a porcine kidney epithelial cell line (LLCPK1), and a human glioma cell line (U-138 MG). The 3,421-bp fragment of the 5'-flanking region resulted in a high level of expression of the reporter gene in all three cell lines. Shorter fragments of the 5'-flanking region resulted in a decrease in the level of CAT reporter expression that varied among the three cell lines, implying the presence of tissue-specific regulatory sites. To study the tissue-specific regulation of the catalase promoter, transgenic mice containing the 3,421-bp 5'-flanking sequence attached to the CAT reporter gene were produced, and CAT expression was measured in various tissues of three independent transgenic lines. CAT activity was consistently high in muscle tissue (heart, skeletal muscle, and diaphragm) and low in most other tissues studied, particularly in liver and kidney. In contrast, the endogenous expression of catalase is low in muscle and high in liver and kidney; thus, the tissue-specific expression of the reporter gene driven by the 3,421-bp fragment of the 5' flanking region of the catalase gene was not similar to the expression of the endogenous catalase gene. PMID- 9397153 TI - Overexpression of HSP25 reduces the level of TNF alpha-induced oxidative DNA damage biomarker, 8-hydroxy-2'-deoxyguanosine, in L929 cells. AB - Previously we and others have demonstrated that oxidative stress involving generation of reactive oxygen species (ROS) is responsible for the cytotoxic action of TNF alpha. Protective effect of small heat shock proteins (HSP) against diverse oxidative stress conditions has been suggested. Although overexpression of small HSP was shown to provide an enhanced survival of TNF alpha-sensitive cells when challenged with TNF alpha, neither the nature of TNF alpha-induced cytotoxicity nor the protective mechanism of small HSP has been completely understood. In this study, we have attempted to determine whether TNF alpha induces oxidative DNA damage in TNF alpha-sensitive L929 cells. We chose to measure the level of 8-hydroxy-2'-deoxyguanosine (8 ohdG), which has been increasingly recognized as one of the most sensitive markers of oxidative DNA damage. Our results clearly demonstrated that the level of 8 ohdG increased in L929 cells in a TNF alpha dose-dependent manner. Subsequently, we asked whether small HSP has a protective effect on TNF alpha-induced oxidative DNA damage. To accomplish this goal, we have stably transfected into L929 cells, which are devoid of endogenous small HSP, with the mouse small hsp cDNA (hsp25). We found that TNF alpha-induced 8 ohdG was decreased in cells overexpressing exogenous small HSP25. We also found that the cell-killing activity of TNF alpha was decreased in these cells as measured by clonogenic survival. Taken together, results from the current study show that a cytotoxic mechanism of TNF alpha involves oxidative damage of DNA, and that overexpression of the small HSP25 reduces this oxidative damage. We suggest that the reduction of oxidative DNA damage is an important protective mechanisms of small HSP against TNF alpha. PMID- 9397155 TI - Selection of highly metastatic rat MTLn2 mammary adenocarcinoma cell variants using in vitro growth response to transferrin. AB - We previously found that the proliferative response to transferrin and the expression of transferrin receptors (TfR) on the cell surface of various rat 13762NF mammary adenocarcinoma cell sublines correlated with their spontaneous metastatic capability. To further assess the involvement of transferrin and TfR in metastasis, transferrin-responsive cells were selected from the poorly metastatic, low-transfferin responsive 13762NF MTLn2 subline. When maintained in low serum (0.3%) conditions, MTLn2 cells failed to survive. However, if like medium was supplemented with 0.5 microgram/ml rat transferrin, some colonies emerged, presumably due to their ability to proliferate in response to the added transferrin. The surviving cells were expanded and exposed to ten or 20 similar cycles of transferrin growth selection to obtain the sublines MTLn2-Tf10 and MTLn2-Tf20, respectively. The MTLn2-Tf20 cells proliferated in response to transferrin at a rate similar to that of the high metastatic 13762NF sublines. Using immunofluorescent staining, Scatchard analysis, and affinity isolation of TfR, we discovered that the MTLn2-Tf20 cells had 5 to 6 times more TfR than did the parental MTLn2 line. When injected into the mammary fat pads of rats, the MTLn2-Tf20 line metastasized to the axillary lymph node in seven out of ten animals and to the lungs in six out of ten (median number = 13). No metastases were seen in the MTLn2 parental line. The MTLn2-Tf10 cells showed intermediate properties compared with the MTLn2 and MTLn2-Tf20 cells. The results indicate that variant cells with a high response to transferrin may be more metastatic than the bulk cells in a poorly metastatic population. The selection of cells with high levels of TfR and a higher proliferative response to transferrin results in sublines with greater potentials for spontaneous metastasis. PMID- 9397154 TI - Repression of mitogen-activated protein kinases ERK1/ERK2 activity by a protein tyrosine phosphatase in rat fibroblasts transformed by upstream oncoproteins. AB - The observation that mitogen-activated protein (MAP) kinases ERK1 and ERK2 are constitutively activated in a number of oncogene-transformed cell lines has led to the hypothesis that prolonged activation of these enzymes is required for the transformation process. To investigate this question, we have examined the regulation of the ERK pathway in Rat1 fibroblasts transformed with activated c Raf-1 (Raf22W), v-Ha-Ras, and v-Src. Expression of these oncoproteins had no effect on the enzymatic activity of ERK1 and ERK2 in either serum-starved or exponentially growing cells. Moreover, the stimulatory effect of serum on ERK1/ERK2 activity was substantially reduced or abrogated in these cells; this impairment was associated with a strong attenuation of c-fos gene induction. In contrast, expression of Raf22w, v-Ha-Ras, or v-Src resulted in the constitutive activation of the upstream kinases MEK1 and MEK2. Treatment of the cells with vanadate completely restored the activation of ERK1/ERK2 in oncogene-transformed cells, suggesting the involvement of a vanadate-sensitive tyrosine phosphatase. Northern blot analysis of VH1-like dual-specificity MAP kinase phosphatases did not reveal any significant difference in the mRNA expression pattern of these genes between parental and transformed Rat1 cells. Phosphoamino acid analysis indicated that ERK1 is phosphorylated on threonine, but not on tyrosine, in oncogene-transformed cells and that vanadate treatment restores tyrosine phosphorylation. We conclude from these results that ERK1/ERK2 activity is repressed by a single-specificity tyrosine phosphatase in oncogene-transformed rat fibroblasts. PMID- 9397156 TI - Un-cross-linked fibrin substrates inhibit keratinocyte spreading and replication: correction with fibronectin and factor XIII cross-linking. AB - Wound repair is characterized by the presence of a fibrin-rich matrix, but the effect of fibrin on re-epithelialization remains unclear. In this study, we determined the effects of different fibrin matrices on cultured human neonatal keratinocytes. Using purified fibrinogen and fibrin gels generated by the enzymatic action of thrombin, batroxobin (it leads to retention of fibrinopeptide B), or Agkistrodon contortrix thrombin-like enzyme (ACTE; it leads to retention of fibrinopeptide A), we determined the effect of each of these matrices on keratinocyte morphology, attachment, spreading, and replication as compared to tissue culture plastic. Morphologically, keratinocytes seeded on fibrin surfaces were more rounded and formed three-dimensional structures. Specific cell attachment, as measured at either 37 degrees C or 4 degrees C, was not altered on the different fibrin substrates (P > .05) but was increased on fibrinogen and factor XIII cross-linked fibrin (P < .01). However, keratinocytes seeded on fibrin, regardless of the presence or absence of fibrinopeptides A or B, showed a marked decrease (up to 71%) in cell numbers by days 5 (P = .0357) and 10 (P = .0114). Keratinocyte spreading was decreased by 78.8% (P = .0006), 80.3% (P = .0001), and 89.2% (P = .0001) on thrombin-, batroxobin-, and ACTE-generated fibrin, respectively, but not on fibrinogen-coated dishes. However, either the addition of fibronectin or cross-linking of fibrin with factor XIII allowed full keratinocyte spreading to occur (P = .0002 and P = .0013, respectively). We conclude that fibrin inhibits keratinocyte spreading in the absence of other matrix or plasma proteins or cross-linking by factor XIII. PMID- 9397157 TI - Rapid disruption of gap junctional communication and phosphorylation of connexin43 by platelet-derived growth factor in T51B rat liver epithelial cells expressing platelet-derived growth factor receptor. AB - Gap junctional communication (GJC) between contacting cells has been postulated to be involved in the regulation of cell proliferation. This suggestion stems from numerous studies showing modulation of GJC by agents that influence cellular proliferation. Platelet-derived growth factor (PDGF), a strong mitogen, inhibits GJC in many cell types. To understand the molecular nature of the signal transduction pathway responsible for the GJC blockade, T51B rat liver epithelial cells, which lack endogenous PDGF receptor (PDGFr), were infected with a retrovirus containing either wild-type full-length cDNA of human PDGFr beta (Kin+) or a mutant PDGFr beta lacking receptor tyrosine kinase activity (Kin-). PDGF caused a complete but transient interruption of cell communication in Kin+ cells within 15-20 min of addition. This interruption of GJC was not associated with a gross destabilization of gap junction plaques but with the phosphorylation of connexin43 (Cx43), the only known gap junction protein expressed in these cells. These effects were exhibited in either control T51B cells or in Kin- cells, indicating a requirement of the receptor tyrosine kinase activity. Further examination revealed that the newly phosphorylated Cx43 then undergoes a rapid degradation utilizing the lysosomal pathway resulting in a decreased total Cx43 protein level. The re-establishment of GJC following PDGF treatment was dependent on protein synthesis. This report describes a suitable cell system which is currently being utilized for the characterization of the PDGF signaling pathway responsible for the inhibition of GJC. PMID- 9397158 TI - Endothelial and serum factors which include apolipoprotein A1 tether elastin to smooth muscle cells inducing serine elastase activity via tyrosine kinase mediated transcription and translation. AB - We previously reported that serine elastase activity is induced in cultured porcine pulmonary artery (PA) smooth muscle cells (SMC) following serum stimulation by a mechanism involving adhesion of elastin to an elastin binding protein and tyrosine kinase activity. The present study demonstrates that a PA endothelial cell factor also promotes a fourfold increase in elastin adhesion to PA SMC and a twofold increase in serine elastase activity. The mechanism involves tethering of the factor to SMC, since [3H]-elastin pre-incubated with serum or endothelial cell (EC)-conditioned medium or SMC pre-treated with serum accelerates binding of elastin and tyrosine-kinase related elastase activity. The serum factor appears to interact with integrins as elastase induction is partially inhibited by RGD peptides. The elastase-inducing properties of serum could not, however, be attributed to several RGD-containing proteins. While a 120 kD fibronectin fragment partially reproduced the effect, it was not found in the serum fraction containing elastase-inducing activity. Instead, a 27 kD serum protein was enriched by elastin affinity chromatography, identified as apolipoprotein (Apo) A1 by microsequence analysis, and found to have about 50% of the elastase-inducing activity of serum. Elastase induction is inhibited by actinomycin and cycloheximide, suggesting a requirement for mRNA transcription and protein synthesis. Our results suggest a novel cell-extracellular matrix interaction whereby a soluble factor, in this case a lipoprotein, binds and tethers a matrix component to the cell surface and induces tyrosine kinase dependent transcription of mRNA culminating in substrate proteolysis. PMID- 9397159 TI - Increased transcription and modified growth state-dependent expression of the plasminogen activator inhibitor type-1 gene characterize the senescent phenotype in human diploid fibroblasts. AB - The type-1 inhibitor of plasminogen activator (PAI-1) is a major physiologic regulator of pericellular proteolytic activity and, as such, influences matrix integrity, cell-to-substrate adhesion, and cellular proliferation. Excessive accumulation of both PAI-1 mRNA and protein correlates with the progressive acquisition of morphological and growth traits characteristic of the senescent phenotype (Mu and Higgins, 1995, J. Cell. Physiol., 165:647-657). Compared to early-passage IMR-90 human diploid fibroblasts, a late-passage senescence associated 11-fold elevation in steady-state PAI-1 mRNA content reflected a 15 fold increase in constitutive PAI-1 gene transcription. Differential mRNA stability was not a factor in age-associated PAI-1 overexpression in IMR-90 cells. Upon removal of serum, early-passage human fibroblasts enter into a state of growth arrest with marked down-regulation of PAI-1 synthesis. Rapid induction of both the 3.0- and 2.2-kb PAI-1 mRNA species was evident upon serum-induced "activation" of quiescent early-passage fibroblasts; induced PAI-1 transcripts were maximal at 2 hr post-serum stimulation and declined in late G1 prior to entry into S phase. In contrast, late-passage (p32) fibroblasts maintained a significant level of PAI-1 expression under serum-free culture conditions. Although the PAI-1 gene was further responsive to serum in senescent cells, transcript abundance remained elevated and actually increased over the 12 to 16 hr post-serum addition period (a time when early-passage fibroblasts down regulate PAI-1 mRNA content). Development of the senescent phenotype in human fibroblasts is associated, therefore, with significant changes in PAI-1 gene regulation. Such reprogramming involves predominantly transcriptional events and results in a marked increase in steady-state PAI-1 transcript abundance involving both the 3.0- and 2.2-kb mRNA species. PMID- 9397160 TI - Retinoic acid-induced differentiation in a human neuroblastoma cell line is associated with an increase in nitric oxide synthesis. AB - The human neuroblastoma cell line SK-N-BE, after incubation with 10 microM retinoic acid (RA) or 20 nM phorbol 12-myristate 13-acetate (PMA), underwent biochemical and morphological signs of differentiation within 10-14 days. In parallel, SK-N-BE cells produced significantly higher amounts of nitric oxide (NO) in comparison with controls, as assessed by the measurement of nitrite and nitrate in the culture supernatant and of NO synthase (NOS) activity in the cell lysates (measured as ability to convert [3H]arginine into [3H]citrulline and as NADPH diaphorase activity). Nitrite/nitrate production was abolished by adding the NO scavenger hemoglobin in the culture medium and was inhibited by aminoguanidine (AG, a selective inhibitor of the inducible NOS isoform) but not by the less selective inhibitor NG-nitro-L-arginine methylester (NAME). Western blotting experiments with monoclonal antibodies against the ncNOS and iNOS isoforms suggest that RA-elicited NOS activation is not attributable to an increased expression of the protein. NAME and AG were not able to revert inhibition of proliferation induced by RA, and the NO donor sodium nitroprusside did not mimic the effect of RA and PMA. These data indicate that increased NO synthesis does not mediate RA- or PMA-induced differentiation but may be an additional marker of differentiation into sympathetic-like neuronal cells. PMID- 9397161 TI - Induction of hepatocyte growth factor/scatter factor by interferon-gamma in human leukemia cells. AB - Induction of hepatocyte growth factor/scatter factor (HGF/SF) may be one of the critical steps in organ regeneration, wound healing, and embryogenesis. We previously reported the production of HGF/SF from various human leukemia cell lines and a high level of the growth factor in blood and bone marrow plasma from patients with various types of leukemia. We determined here the effects of hematopoietic cytokines on HGF/SF production in human leukemia cell lines, KG-1, a myeloid cell line, and RPMI-8226, a B cell line. Interferon (IFN)-gamma remarkably stimulated HGF/SF production in both cell lines at concentrations of more than 0.1 or 1 IU/ml. IFN-alpha and IFN-beta were as effective as IFN-gamma in RPMI-8226 cells, but less than IFN-gamma in KG-1 cells. HGF/SF gene expression in KG-1 cells was also up-regulated by IFN-gamma. Granulocyte colony-stimulating factor (G-CSF), granulocyte/macrophage colony-stimulating factor (GM-CSF), interleukin (IL)-5 and IL-6 had no effect on HGF/SF production in the 2 leukemia cell lines. We also determined the effects of HGF/SF inducers known for human fibroblasts on the growth factor production in leukemia cells. Out of phorbol 12 myristate 13-acetate (PMA), cholera toxin, IL-1 beta, and tumor necrosis factor (TNF)-alpha, the former three were as effective as IFN-gamma in KG-1 cells, but only TNF-alpha stimulated HGF/SF production in RPMI-8226 cells, whose effect was less than those of IFN-alpha, IFN-beta, and IFN-gamma. The effect of IFN-gamma in KG-1 cells was synergistic with that of PMA. In contrast with the effect in leukemia cells, HGF/SF induction by IFN-gamma in human skin fibroblasts was much less than that by PMA or cholera toxin. These results indicated that IFN-gamma is a potent inducer of HGF/SF in human leukemia cells. This finding suggests the presence of a homeostatic control mechanism in liver regeneration and repair: hepatic injury, DNA synthesis inhibition, or apoptosis caused by IFN-gamma is subsequently overcome by cytokine-induced HGF/SF, a potent promoter of liver DNA synthesis. PMID- 9397162 TI - Co-regulation of pituitary tumor cell adhesion and prolactin gene expression by glucocorticoid. AB - Rat 235-1 pituitary tumor cells are lactotrophs producing high levels of prolactin (PRL). Dexamethasone (Dex, 100 nM) inhibits PRL gene expression in 235 1 cells by 50%, while simultaneously decreasing cell replication and cell-cell aggregation. To determine the time course of Dex action, we used a quantitative assay for cell-cell interaction, based on the number of single cells present before and after re-aggregation of dispersed cells. 235-1 cells were cultured in growth medium or medium plus 100 nM Dex for 1-4 days before assay. Control cells had 90% re-aggregation on all days of assay. Aggregation of Dex-treated cells decreased to 55% by day 4. Dex treatment also reduced cell numbers by 40%, but this decrease did not contribute to reduced aggregation. To determine the mechanism of Dex-inhibited cell-cell adhesion, we examined the expression of cadherins and catenins. Cadherin-related mRNAs (P- and N-cadherin probes) were detectable in 235-1 cells, but their levels were unchanged by Dex. A pancadherin antibody was unable to detect classical cadherins in these cells. Both alpha- and beta-catenins were detected by Western blotting and their levels were decreased by Dex. Unlike control aggregates, aggregates of Dex-treated cells were able to inhibit expression of PRL mRNA when added to monolayers of 235-1 cells. These data suggest that Dex influences cadherin function by inhibiting catenin expression and that this has the functional consequence of altering 235-1 cell cell interactions. Overall the data show that Dex affects important aspects of lactotroph function other than PRL gene expression. These changes may include physical alterations in pituitary cell contacts that further support a change in functional state. PMID- 9397163 TI - Regulation of spermidine/spermine N1-acetyltransferase expression by cytokines and polyamines in human hepatocarcinoma cells (HepG2). AB - Spermidine/spermine N1-acetyltransferase (cSAT), a key enzyme in polyamine degradation, is induced by various hepatotoxins and liver tumor promoters. In this paper we demonstrate that physiological factors, such as cytokines, control cSAT expression in HepG2 human hepatocarcinoma cells. Hepatocyte growth factor (HGF) induced the cSAT mRNA precursor (3.5 kb) at 4 h. The mature form of mRNA (1.3 kb) increased 6-8-fold between 8 and 10 h, and remained elevated until 18 h. An increase in cSAT activity (2-fold) and high levels of N1-acetylspermidine were observed concomitantly. Interleukin-1 beta (IL-1 beta) enhanced cSAT expression (both mRNA and enzyme activity) similar to HGF, while tumor necrosis factor-alpha (TNF-alpha) was less effective. This system also provides a useful means for examining the involvement of negative and positive changes of polyamines in the induction of cSAT and c-jun, a gene that participates in the control of cSAT expression. alpha-Difluoromethylornithine (DFMO) pretreatment, by lowering putrescine and spermidine in HGF- or IL-1 beta-treated cells, prevented the induction of cSAT. This effect was reversed by exogenous putrescine or spermidine. IL-1 beta induced c-jun mRNA more than HGF. DFMO prevented almost completely the enhancement of c-jun mRNA expression by IL-1 beta, and this effect was reversed by exogenous putrescine or spermidine. Therefore, we suggest that cSAT and c-jun expression is specifically regulated by polyamine-mediated mechanisms in IL-1 beta treated HepG2 cells. Since cSAT is inducibile by cytokines that control tumor metabolism and growth as well as tumor-host interaction, we hypothesize an involvement of cSAT in hepatoma growth. PMID- 9397164 TI - Potent inhibition of cell density-dependent apoptosis and enhancement of survival by dimethyl sulfoxide in human myeloblastic HL-60 cells. AB - Human myeloblastic cell line HL-60 cells undergo apoptosis during in vitro culture in a cell density-dependent manner, and this cell density-dependent apoptosis was observed when the concentration of cultured cells exceeded 8-10 x 10(5) cells/ml. Dimethyl sulfoxide (DMSO), a differentiation inducer of HL-60 cells, did not amplify, but rather potently inhibited, this apoptosis. In a low density culture condition, DMSO attenuated proliferation of HL-60 cells in spite of its inhibition of apoptosis. In contrast, DMSO did support cell survival under high cell density conditions, and DMSO-treated HL-60 cells reached an extremely high concentration of 2-3 x 10(6) cells/ml, a condition which could never be possible in a usual culture environment. Thus, DMSO exerted dual effects on cell proliferation, i.e., growth inhibition and apoptosis inhibition, and the sum of these effects resulted in an apparently distinct phenomenon according to the culture conditions including cell density. PMID- 9397165 TI - 3 Beta-(4-ethyl-3-iodophenyl)nortropane-2 beta-carboxylic acid methyl ester as a high-affinity selective ligand for the serotonin transporter. PMID- 9397167 TI - Potent and selective inhibition of neuronal nitric oxide synthase by N omega propyl-L-arginine. PMID- 9397166 TI - Class III antiarrhythmic activity in vivo by selective blockade of the slowly activating cardiac delayed rectifier potassium current IKs by (R)-2-(2,4 trifluoromethyl)-N-[2-oxo-5-phenyl-1-(2,2,2-trifluoroethyl)- 2, 3-dihydro-1H benzo[e][1,4]diazepin-3-yl]acetamide. PMID- 9397168 TI - A new approach to docking in the beta 2-adrenergic receptor that exploits the domain structure of G-protein-coupled receptors. AB - A novel technique for docking ligands to the beta 2-adrenergic receptor is described which exploits the domain structure of this class of receptors. The ligands (norepinephrine, an agonist; pindolol, a partial agonist; and propranolol, an antagonist) were docked into the receptor using the key conserved aspartate on helix 3 (D113) as an initial guide to the placement of the amino group and GRID maps (Goodford, P. J. J. Med. Chem, 1985, 28, 849) to identify the likely binding regions of the hydrophobic (and hydroxyl) moieties on the A domain (comprising of helices 1-5). The essence of the new approach involved pulling the B domain, which includes helices 6 and 7, away from the other domain by 5-7 A. During the subsequent minimization and molecular dynamics, the receptor ligand complex reformed to yield structures which were very well supported by site directed mutagenesis data. In particular, the model predicted a number of important interactions between the antagonist and key residues on helix 7 (notably Leu311 and Asn312) which have not been described in many previous computer simulation studies. The justification for this new approach is discussed in terms of (a) phase space sampling and (b) mimicking the natural domain dynamics which may include domain swapping and dimerization to form a 5,6-domain swapped dimer. The observed structural changes in the receptor when pindolol, the partial agonist, was docked were midway between those observed for propranolol and norepinephrine. These structural changes, particularly the changes in helix helix interactions at the dimer interface, support the idea that the receptors have a very dynamic structure and may shed some light on the activation process. The receptor model used in these studies is well supported by experiment, including site-directed mutagenesis (helices 1-7), zinc binding studies (helices 2, 3, 5, and 6), the substituted cysteine accessibility method (helices 3, 5, and 7), and site-directed spin-labeling studies (helices 3-6). PMID- 9397170 TI - Syntheses and photodynamic activities of novel trisulfonated zinc phthalocyanine derivatives. AB - The synthesis of water-soluble, unsymmetrical, trisulfonated zinc phthalocyanines (ZnPcS3) as single products of the ring expansion of boron tri(4 sulfo)subphthalocyanine (SubPc) is reported. The novel, water-soluble trisulfo SubPcB(OH) was prepared via hydrolysis of the tris(4-chlorosulfonyl)SubPcB(Br) which in turn was obtained from the condensation of 4 (chlorosulfonyl)phthalonitrile with BBr3 in 1-chlorobenzene. A number of ZnPcS3 analogues were prepared via the reaction of S3SubPcB (OH) with different diiminoisoindoline derivatives of increasing hydrophobicity. The reaction proceeds at relative low temperature with acceptable yields. Metalation of free base Pc's with zinc acetate dihydrate afforded the corresponding zinc complexes. Photodynamic activities were measured against the EMT-6 mouse mammary tumor cell line and compared to those of the known ZnPcS3 and ZnPcS4. Added (t-Bu)benzo and (t-Bu)naphtho groups increased the in vitro cell photoinactivation efficacy of the ZnPcS3, whereas addition of a fourth sulfobenzo or bulky diphenylpyrazino group decreased the activity of the parent molecule. The (t Bu)naphthotrisulfobenzoporphyrazine induced the best in vivo photodynamic tumor control which, combined with its good solubility and broad absorption spectrum, renders this compound an interesting dye for photodynamic applications in medicine. PMID- 9397169 TI - Boron-containing polyamines as DNA targeting agents for neutron capture therapy of brain tumors: synthesis and biological evaluation. AB - Three series of new boron-containing spermidine/spermine (SPD/SPM) analogues have been synthesized: N1- and N5-(4-carboranylbutyl) SPD/SPM derivatives (SPD-1, SPD 5, SPM-1, SPM-5); N1,N10-diethyl-N5-(4-carboranylbutyl)spermidine (DESPD-5), N1,N14-diethyl-N5-(4-carboranylbutyl)spermine (DESPM-5); and N5,N10-bis(4 carboranylbutyl)spermine (SPM-5,10). In vitro studies using rat F98 glioma cells have shown that these polyamines retain the ability to displace ethidium bromide from calf thymus DNA and are rapidly taken up by F98 glioma cells. However, their cytotoxicities, especially those with terminal N-substituted (SPD-1, SPM-1) boron compounds, are greater than those of SPD/SPM. Nevertheless, the groundwork has been created for a new class of boron-containing compounds that maybe useful for boron neutron capture therapy of tumors. PMID- 9397171 TI - N-hydroxyalkyl derivatives of 3 beta-phenyltropane and 1-methylspiro[1H-indoline 3,4'-piperidine]: vesamicol analogues with affinity for monoamine transporters. AB - As part of our ongoing structure-activity studies of the vesicular acetylcholine transporter ligand 2-(4-phenylpiperidino)cyclohexanol (vesamicol, 1), 22 N hydroxy(phenyl)alkyl derivatives of 3 beta-phenyltropane, 6, and 1-methylspiro[1H indoline-3,4'-piperidine], 7, were synthesized and tested for binding in vitro. Although a few compounds displayed moderately high affinity for the vesicular acetylcholine transporter, no compound was more potent than the prototypical vesicular acetylcholine transporter ligand vesamicol. However, a few derivatives of 6 displayed higher affinity for the dopamine transporter than cocaine. We conclude that modification of the piperidyl fragment of 1 will not lead to more potent vesicular acetylcholine transporter ligands. PMID- 9397172 TI - Tyrosine kinase inhibitors. 13. Structure-activity relationships for soluble 7 substituted 4-[(3-bromophenyl)amino]pyrido[4,3-d]pyrimidines designed as inhibitors of the tyrosine kinase activity of the epidermal growth factor receptor. AB - The general class of 4-(phenylamino)quinazolines are potent (some members with IC50 values << 1 nM) and selective inhibitors of the tyrosine kinase activity of the epidermal growth factor receptor (EGFR), via competitive binding at the ATP site of the enzyme, but many of the early analogues had poor aqueous solubility (<< 1 mM). A series of 7-substituted 4-[(3-bromophenyl)-amino]pyrido[4,3 d]pyrimidines, together with selected (3-methylphenyl)amino analogues, were prepared by reaction of the analogous 7-fluoro derivatives with appropriate amine nucleophiles in 2-BuOH or aqueous 1-PrOH. All of the compounds were evaluated for their ability to inhibit the tyrosine-phosphorylating action of EGF-stimulated full-length EGFR enzyme. Selected analogues were also evaluated for their inhibition of autophosphorylation of the EGF receptor in A431 human epidermoid carcinoma cells in culture and against A431 tumor xenografts in mice. Analogues bearing a wide variety of polyol, cationic, and anionic solubilizing substituents retained activity, but the most effective in terms of both increased aqueous solubility (> 40 mM) and retention of overall inhibitory activity (IC50's of 0.5 10 nM against isolated enzyme and 8-40 nM for inhibition of EGFR autophosphorylation in A431 cells) were weakly basic amine derivatives. These results are broadly consistent with a proposed model for the binding of these compounds to EGFR, in which the 6- and 7-positions of the pyridopyrimidine ring are in a largely hydrophobic binding region of considerable steric freedom, at the entrance of the adenine binding cleft. The most active cationic analogues have a weakly basic side chain where the amine moiety is three or more carbon atoms away from the nucleus. Two of the compounds (bearing weakly basic morpholinopropyl and strongly basic (dimethylamino)butyl solubilizing groups) produced in vivo tumor growth delays of 13-21 days against advanced stage A431 epidermoid xenografts in nude mice, when administered i.p. twice per day on days 7-21 posttumor implant. Treated tumors did not increase in size during therapy and resumed growth at the termination of therapy, indicating an apparent cytostatic effect for these compounds under these treatment conditions. The data suggest that continuous long-term therapy with these compounds may result in substantial tumor growth inhibition. PMID- 9397173 TI - New methodology for profiling combinatorial libraries and screening sets: cleaning up the design process with HARPick. AB - Combinatorial chemistry is a tool of increasing importance in the field of ligand design, as it can yield huge increases in the number of compounds available for screening. Unfortunately, it is often the case that the number of molecules which could theoretically be constructed greatly exceeds potential synthesis and screening capacity. For this new technology to be fully exploited, it will become vital to design libraries with reference to the properties of compounds already in existence, if the added value of each new molecular collection is truly to be maximized. Similarly, if we are to take full advantage of the potential of combinatorial chemistry in lead optimization, it is important that our library design paradigms are flexible, with diversity scoring functions that can be modified to suit particular projects. Here these challenges are addressed through the introduction of a novel computer-aided library design tool known as HARPick (heuristic algorithm for reagent picking). The program is accessible to the bench chemist, and incorporates several significant advances over currently available approaches. These include product-based diversity calculations that can be constrained at the reagent level; diversity measures constructed from multiple descriptors; improved pharmacophore key information and full pharmacophore profiling of entire molecular databases. The potential of these improvements to aid in diversity profiling is illustrated through comparison with established methodology, and possible further enhancements are discussed. PMID- 9397174 TI - Antirhino/enteroviral vinylacetylene benzimidazoles: a study of their activity and oral plasma levels in mice. AB - In an effort to find an orally bioavailable antiviral for the treatment of rhino/enteroviral infections, a series of vinylacetylene benzimidazoles (11a-o, 12, and 18a) was made. Initial studies of this class of antivirals showed that fluorine substitution on the left-hand phenyl ring in combination with the vinylacetylene moiety gave the requisite mix of physical properties to achieve good in vitro antiviral activity as well as respectable oral bioavailability in rhesus monkeys. To ascertain the generality of this finding and to broaden the scope of the structure-activity relationship (SAR), the present study concentrated on fluoro substitution of this class of molecules. The initial antiviral activity for each analogue was measured using human rhinovirus 14 (HRV 14). This served as an indicator of general antiviral activity for SAR purposes. Subsequently, the spectrum of antirhino/enteroviral activity of the more interesting analogues was evaluated through testing against a panel of seven additional rhino/enteroviruses. Broad-spectrum activity was present and consistent for all analogues tested, and it tracked closely with the antiviral activity observed against HRV-14. A simple screening protocol for oral bioavailability was established whereby compounds were administered orally to mice and plasma levels were measured. This procedure facilitated the evaluation of numerous analogues in a rapid manner. The Cmax was used as a measure of oral bioavailability to allow relative ranking of compounds. In general, fluorine substitution directly on the left-hand aromatic ring does give good oral blood levels. However, fluorine incorporation at other positions in the molecule was not as effective at maintaining either the activity or the oral plasma levels. The constructive combination of activity and oral plasma levels was maximized in three derivatives: 11a,e,g. PMID- 9397175 TI - Synthesis and biological properties of new constrained CCK-B antagonists: discrimination of two affinity states of the CCK-B receptor on transfected CHO cells. AB - To improve our knowledge of the bioactive conformation of CCK-B antagonists, we have developed a new series of constrained dipeptoids whose synthesis and biochemical properties are reported here. These compounds, of general structure N alpha-[(2-adamantyloxy)carbonyl]-alpha-methyltryptophanyl-(4 -X)-proline, were designed by introducing a cyclization in the structure of the previously described CCK-B/peptoid antagonist RB 210, N-[N-[(2-adamantyloxy)carbonyl]-DL alpha-methyltryptophanyl] -N-(2-phenylethyl)glycine (Blommaert et al. J. Med. Chem. 1993, 36, 2868-2877), by means of a five-membered ring. Structure-affinity relationship studies showed that an R configuration of Trp-C alpha and a cis configuration of the pyrrolidine substituents were favorable for receptor recognition. The most potent compounds of this new series had similar affinities for the CCK-B receptor as RB 210 and proved to be far more efficient in inhibiting inositol phosphate production in CHO cells stably transfected with rat brain CCK-B receptor, with IC50 values approaching those of the commonly used antagonists L-365,260 and PD-134,308. Moreover, binding studies performed using transfected CHO cells showed that two affinity states of the CCK-B receptor can be discriminated by some of these compounds which also have different biological profiles and are therefore highly interesting tools for the biochemical and pharmacological characterization of CCK-B receptor heterogeneity. PMID- 9397176 TI - Cyclic enkephalin analogues with exceptional potency and selectivity for delta opioid receptors. AB - Superpotent and highly delta-opioid receptor selective cyclic peptides of the general formula H-Tyr-c[D-Pen-Gly-Phe(p-X)-Pen]-Phe-OH (where X = hydrogen or halogen) have been synthesized. In the binding assays the most selective and most potent compound is the p-bromophenyl-alanine-4 analogue (IC50 value = 0.19 nM, selectivity ratio = 21,000 for delta vs mu). In the GPI and MVD bioassays the most selective and most potent analogue is the p-fluoro-substituted analogue Tyr [D-Pen-Gly-Phe(p-F)-Pen]-Phe-OH. In the MVD assay it has an exceptionally low IC50 value of 0.016 nM and a delta vs mu selectivity ratio of 45,000. PMID- 9397177 TI - NAcSDKP analogues resistant to angiotensin-converting enzyme. AB - Two series of analogues of the tetrapeptide NAcSDKP, an inhibitor of hematopoietic stem cell proliferation, were prepared, and their enzymatic stability toward rabbit lung angiotensin-converting enzyme (ACE) was evaluated as well as their capacity to inhibit NAcSDKP hydrolysis by this enzyme. In the first series, each of the peptide bonds has been successively replaced by an aminomethylene bond. In the second one, the C-terminus of the peptide has been modified by decarboxylation or amidation. The results reported here indicate that all of these molecules but one have good stability toward the enzyme but none of the compounds is able to inhibit NAcSDKP hydrolysis by ACE. PMID- 9397178 TI - Enzymatic properties of the unnatural beta-L-enantiomers of 2',3' dideoxyadenosine and 2',3'-didehydro-2',3'-dideoxyadenosine. AB - The beta-L-enantiomers of 2',3'-dideoxyadenosine and 2',3'-didehydro-2',3' dideoxyadenosine have been stereospecifically synthesized. In an attempt to explain the previously reported antiviral activities of these compounds, their enzymatic properties were studied with respect to adenosine kinase, deoxycytidine kinase, adenosine deaminase, and purine nucleoside phosphorylase. Adenosine deaminase was strictly enantioselective and favored beta-D-ddA and beta-D-d4A, whereas adenosine kinase and purine nucleoside phosphorylase had no apparent substrate properties for the D- or L-enantiomers of beta-ddA or beta-d4A. Human deoxycytidine kinase showed a remarkable inversion of the expected enantioselectivity, with beta-L-ddA and beta-L-d4A having better substrate efficiencies than their corresponding beta-D-enantiomers. Our results demonstrate the potential of beta-L-adenosine analogues as antiviral agents and suggest that deoxycytidine kinase has a strategic importance in their cellular activation. PMID- 9397179 TI - 5-HT1B receptor antagonist properties of novel arylpiperazide derivatives of 1 naphthylpiperazine. AB - A new series of arylpiperazide derivatives of 1-naphthylpiperazine of general formula 4 has been prepared and evaluated as 5-HT1B antagonists. Binding experiments at cloned human 5-HT1A, 5-HT1B, and 5-HT1D receptors show that these derivatives are potent and selective ligands for 5-HT1B/1D subtypes with increased binding selectivity versus the 5-HT1A receptor when compared to 1 naphthylpiperazine (1-NP). Studies of inhibition of the forskolin-stimulated cAMP formation mediated by the human 5-HT1B receptor demonstrate that the nature of the arylpiperazide substituent modulates the intrinsic activity of these 1-NP derivatives. Among them, 2-[[8-(4-methylpiperazin-1-yl)naphthalen-2-yl]oxy] -1-(4 o-tolylpiperazin-1-yl)ethanone (4a) was identified as a potent neutral 5-HT1B antagonist able to antagonize the inhibition of 5-HT release induced by 5-CT (5 carbamoyltryptamine) in guinea pig hypothalamus slices. Moreover, 4a was found to potently antagonize the hypothermia induced by a selective 5-HT1B/1D agonist in vivo in the guinea pig following oral administration (ED50 = 0.13 mg/kg). PMID- 9397180 TI - Viracept (nelfinavir mesylate, AG1343): a potent, orally bioavailable inhibitor of HIV-1 protease. AB - Using a combination of iterative structure-based design and an analysis of oral pharmacokinetics and antiviral activity, AG1343 (Viracept, nelfinavir mesylate), a nonpeptidic inhibitor of HIV-1 protease, was identified. AG1343 is a potent enzyme inhibitor (Ki = 2 nM) and antiviral agent (HIV-1 ED50 = 14 nM). An X-ray cocrystal structure of the enzyme-AG1343 complex reveals how the novel thiophenyl ether and phenol-amide substituents of the inhibitor interact with the S1 and S2 subsites of HIV-1 protease, respectively. In vivo studies indicate that AG1343 is well absorbed orally in a variety of species and possesses favorable pharmacokinetic properties in humans. AG1343 (Viracept) has recently been approved for marketing for the treatment of AIDS. PMID- 9397181 TI - Effect of stereochemistry on the clearance mechanism of 111In(III)-labeled D- or L-benzyldiethylenetriaminepentaacetic acid. AB - The diethylenetriaminepentaacetic acid (DTPA) derivatives L-Bn-DTPA and D-Bn-DTPA were synthesized and radiolabeled with 111In3+. The uptake and clearance of the compounds were determined through biodistribution and excretion studies in Wistar rats. Both isomers readily cleared from the animal. The D isomer showed relatively high kidney uptake and predominantly renal clearance. The L isomer showed substantial kidney and liver uptake with equal biliary and renal clearance. Clearance was also evaluated in TR- Wistar rats, which are defective in the liver canalicular multispecific organic anion transporter (cMOAT) protein. cMOAT mediates hepatobiliary clearance of many organic anions. Both compounds were excreted through the urine in TR- Wistar rats, suggesting that cMOAT is important in the clearance of the compounds from the liver. PMID- 9397182 TI - Investigation of the potential impact of benchmark dose and pharmacokinetic modeling in noncancer risk assessment. AB - There has been relatively little attention given to incorporating knowledge of mode of action or of dosimetry of active toxic chemical to target tissue sites in the calculation of noncancer exposure guidelines. One exception is the focus in the revised reference concentration (RfC) process on delivered dose adjustments for inhaled materials. The studies reported here attempt to continue in the spirit of the new RfC guidelines by incorporating both mechanistic and delivered dose information using a physiologically based pharmacokinetic (PBPK) model, along with quantitative dose-response information using the benchmark dose (BMD) method, into the noncancer risk assessment paradigm. Two examples of the use of PBPK and BMD techniques in noncancer risk assessment are described: methylene chloride, and trichloroethylene. Minimal risk levels (MRLs) based on PBPK analysis of these chemicals were generally similar to those based on the traditional process, but individual MRLs ranged from roughly 10-fold higher to more than 10-fold lower than existing MRLs that were not based on PBPK modeling. Only two MRLs were based on critical studies that presented adequate data for BMD modeling, and in these two cases the BMD models were unable to provide an acceptable fit to the overall dose-response of the data, even using pharmacokinetic dose metrics. A review of 10 additional chemicals indicated that data reporting in the toxicological literature is often inadequate to support BMD modeling. Three general observations regarding the use of PBPK and BMD modeling in noncancer risk assessment were noted. First, a full PBPK model may not be necessary to support a more accurate risk assessment; often only a simple pharmacokinetic description, or an understanding of basic pharmacokinetic principles, is needed. Second, pharmacokinetic and mode of action considerations are a crucial factor in conducting noncancer risk assessments that involve animal to-human extrapolation. Third, to support the application of BMD modeling in noncancer risk assessment, reporting of toxicity results in the toxicological literature should include both means and standard deviations for each dose group in the case of quantitative endpoints, such as relative organ weights or testing scores, and should report the number of animals affected in the case of qualitative endpoints. PMID- 9397183 TI - Comparison of the binding potential of various diisocyanates on DNA in vitro. AB - Inhalation of diisocyanate vapors is associated with immediate-type hypersensitivity reactions and direct toxic responses. The genotoxic effects of diisocyanates have not been clarified. The aim of this study was to examine the changes in DNA following in vitro exposure to three most commonly used diisocyanates (toluene diisocyanate, TDI; methylenediphenyl-4,4'-diisocyanate, MDI; and hexamethylene diisocyanate, HDI) and to compare their binding potential using melting behavior of DNA and electrophoresis studies in DNA. Following incubation of DNA with MDI (pure and mix) and HDI we found no differences in the melting behavior compared to the control calf thymus DNA. However, DNA treated with TDI showed differences in the shape of the native DNA curves due to changes in hyperchromicity and exhibited 14% more DNA reconstitution after renaturation. The small changes in the melting behavior of native DNA do not suggest the formation of DNA intrastrand cross-links but rather conformational changes of single- and double-stranded DNA. These conformational changes were further explored by agarose electrophoresis of native and denatured calf thymus DNA. Control and all diisocyanate-exposed DNA showed no differences in the size of native DNA fragments. Conversely, electrophoresis of TDI mix-incubated DNA, following denaturation, showed a distinct reduction in the double-stranded DNA fragment size compared to the control, MDI-denatured (pure and mix), and HDI denatured DNA. These findings may help to better understand the mechanisms of the genotoxic effect of TDI. PMID- 9397184 TI - Cadmium toxicity and distribution in metallothionein-I and -II deficient transgenic mice. AB - To date, numerous correlative studies have implicated metallothionein in the detoxification of heavy metals and in the regulation of metal distribution within an organism. In the present study cadmium-binding proteins (metallothionein equivalents), cadmium acute toxicity, and cadmium distribution in tissues and subcellular fractions were compared in metallothionein-I and -II deficient (MT-/ ) mice and the parental strain carrying intact metallothionein genes (MT+/+) to determine if the absence of metallothionein altered any of these parameters. In an uninduced state, MT-/- mice expressed lower levels of cadmium-binding proteins relative to MT+/+ mice in several tissues. Administration of zinc enhanced the levels of cadmium-binding proteins in liver, small intestine, kidney, pancreas, and male sex organs, but not in cecum or brain of MT+/+ mice compared to zinc pretreated MT-/- mice. The cadmium LD50 was similar for MT-/-, MT+/+, and zinc pretreated MT-/- mice (15-17 mumol CdCl2/kg body weight delivered i.p.). However, zinc-pretreated MT+/+ mice had a cadmium LD50 of 58-63 mumol CdCl2/kg body weight. Over two-thirds of cadmium was found in liver, cecum, small intestine, and kidney in both MT+/+ and MT-/- mice; therefore, metallothionein levels do not appear to play a major role in the tissue distribution of cadmium. However, after zinc pretreatment, MT+/+ mice accumulated more cadmium in the liver and less in other tissues, whereas the amount of cadmium in the liver was not altered by zinc pretreatment in MT-/- mice. In general, the cytosolic/particulate ratio of cadmium was significantly higher in tissues of noninduced MT+/+ mice relative to MT-/- mice. This difference was accentuated after zinc pretreatment. Together these results indicate that basal levels of metallothionein do not protect from the acute toxicity of a single i.p. cadmium challenge. Furthermore, it does not appear that the cytosolic compartmentalization of cadmium is correlated with reduced toxicity. PMID- 9397185 TI - Alterations of male Wistar rat jejunum induced by Dodine (n-dodecylguanidine acetate). AB - The effect of Dodine on the intestine was studied after a single administration of 1000 mg/kg, which corresponds to the LD50 in male Wistar rats. At this dose, a significant decrease in body weight was observed, accompanied by diarrhea, which may be associated with intestinal alterations. The chemical induced a significant reduction of the protein content and in sucrase activity in the jejunum. Morphological alterations included a significant decrease in crypt height and in villus length and depth. The intestinal modifications observed in animals after Dodine administration may explain the observed loss in body weight and diarrhea. PMID- 9397186 TI - Gene knockout and transgenic technologies in risk assessment: the next generation. AB - Transgenic and knockout mice have been proposed as substitutes for one of the standard 2-yr rodent assays. The advantages of using genetically engineered mouse models is that fewer mice are needed, the time to develop disease is greatly reduced, and the mice are predisposed to developing cancer by virtue of gain or loss of functions. The models currently being used have yielded a large amount of data and have proved to be informative for risk assessment; however, they are still far from ideal. In fact, they inherently do not reflect the complexity of mutation and carcinogenesis in humans. Recent advances in technology and the creation of new knockout mice may produce more useful and more sensitive models. This review covers two recent advances in technology--inducible and regulatable gene expression and targeted genetic modifications in the genome--that will allow us to make better models. I also discuss new gene deletion and transgenic mouse models and their potential impact on risk-assessment assays. These models are presented in the context of four basic components or events that occur in the multistep process leading to cancer: maintenance of gene expression patterns, genome stability and DNA repair, cell-cell communication and signaling, and cell cycle regulation. Finally, surrogate markers and utility in risk assessment are also discussed. This review is meant to stimulate further discussion in the field and to generate excitement about working toward the next generation of risk assessment models. PMID- 9397187 TI - New directions for predicting carcinogenesis. AB - Carcinogenicity testing today normally includes conducting 2-yr studies of rats and mice of both sexes and following widely accepted procedures for husbandry, selection of dose levels, pathology and toxicity observations, and statistical interpretation of tumor data. These studies are usually preceded by tests for genetic toxicity and subchronic toxicity studies to select dose levels for the 2 yr studies. While these data are used for quantitative risk assessment, the mechanistic basis for effects is usually unknown, and such series of studies are very expensive and require five or more years to conduct. Alternate approaches are being developed that would provide more mechanistic information and perhaps would permit decisions to be made about carcinogenic potential without the need to conduct 2-yr studies of rats and mice of both sexes. Decisions could be based on a profile of data rather than the result of one test. Regulatory acceptance of new approaches for carcinogenicity testing is critical to future progress in the field of carcinogenesis. PMID- 9397188 TI - Potency grading in carcinogen classification. AB - In 1992 the United Nations Conference on Environment and Development decided to harmonize carcinogen classification systems. A proposal for a harmonized classification system is currently being considered by the Organization for Economic Cooperation and Development (OECD). In many countries, classification of a chemical as carcinogenic triggers labeling requirements. Implicit in the labeling requirements are often restrictions on the sale of consumer products and workplace regulations. Many of the current classification systems for carcinogens use a single concentration limit for the minimum concentration of a carcinogen in a preparation (mixture) that requires labeling. For high-potency carcinogens, one concentration limit may not adequately express the hazard, whereas for low potency carcinogens, one limit may overestimate the hazard caused by the carcinogen in the preparation (mixture). The potency grading system discussed consists of three potency groups: high-, medium-, and low-potency carcinogens. It is envisioned that the different classes will trigger different labeling requirements. In the process of potency grading, a preliminary conclusion as to whether a substance shows high, medium, or low potency is initially based on a tumorigenic dose descriptor. The preliminary potency evaluation may then be modified after due consideration of a number of additional elements. These may include evaluation of the dose-response curve; site-, species-, strain-, and sex specific activity; mechanisms including genotoxicity; mechanistic relevance to humans; toxicokinetics; and other factors. The potency grading system discussed is applicable to most carcinogen classification systems, including that currently being considered by the OECD. PMID- 9397189 TI - Differential display identified changes in mRNA levels in regenerating livers from chloroform-treated mice. AB - The nongenotoxic-cytotoxic carcinogen chloroform induces liver necrosis, regenerative cell proliferation, and, eventually, liver tumors in female B6C3F1 mice when administered by gavage at doses of 238 or 477 mg/kg/d. Administration of 1800 ppm of chloroform in the drinking water results in similar daily doses but does not produce liver toxicity or cancer. The differential-display technique was used to compare the expression of a subset of mRNAs in normal (control) and regenerating liver after chloroform-induced toxicity to define the proportion of genes whose expression changes under hepatotoxic conditions and to identify the genes that might play a role in regeneration and perhaps cancer. RNA was purified from the livers of female B6C3F1 mice after 4 d or 3 wk of gavage treatment with 3, 238, or 477 mg/kg/d of chloroform or treatment with 1800 ppm chloroform in drinking water. There was a remarkably high degree of consistency of gene expression among the animals and across dose and treatment groups as visualized by the differential-display technique. Of the 387 bands observed, only four (about 1%) changed expression in regenerating liver. The genes were assigned locus names by GenBank after sequence submission. The genes with increased mRNA levels as confirmed by northern blot analysis were MUSTIS21, a mouse primary response gene induced by growth factors and tumor promoters; MUSMRNAH, a gene highly homologous to a human gene isolated from a prostate carcinoma cell line; and MUSFRA, a novel gene. The novel gene MUSFRB exhibited decreased mRNA levels. No change in expression was seen among control mice given the nontoxic regimens of 3 mg/kg/d chloroform or 1800 ppm chloroform in drinking water, indicating that changes in expression were associated with toxicity and regeneration rather than chloroform per se. These genes and others that may be identified by expanding this approach may play a role in regeneration and perhaps in the process of chloroform-induced carcinogenesis in rodent liver. PMID- 9397190 TI - Susceptibility of transgenic mice carrying human prototype c-Ha-ras gene in a short-term carcinogenicity study of vinyl carbamate and ras gene analyses of the induced tumors. AB - To determine if hemizygous transgenic mice carrying the human c-Ha-ras gene (CB6F1-Tg Hras2 mice (Hras2 mice)) are susceptible to the carcinogenic potential of known murine carcinogens, male and female Hras2 mice and their non-transgenic CB6F1 littermates (non-Tg mice) were each given a single intraperitoneal injection of 60 mg of vinyl carbamate (VC)/kg body weight or saline (vehicle control) and monitored for 16 wk without further treatment. At necropsy, grossly visible tumors were fixed for histopathologic diagnosis and, when of sufficient size, portions were frozen for subsequent molecular analysis. Nine of 31 male and nine of 29 female Hras2 mice treated with VC died within 16 wk as a result of lung tumor burden. At the termination of the study, lung tumors (alveolar bronchiolar epithelial neoplasms and hemangiosarcomas) and focal alveolar bronchiolar hyperplasias were present in both sexes of Hras2 and non-Tg mice treated with VC; there were significantly more proliferative lung lesions in Hras2 than non-Tg mice. Splenic hemangiosarcomas and squamous cell tumors of the forestomach were induced in male and female VC-treated Hras2 mice but not in VC treated non-Tg mice. Polymerase chain reaction-single-strand conformation polymorphism analysis and DNA sequencing of the induced lung tumors revealed point mutations at codon 61 of the transgene in two of 29 lung tumors (one of 16 in males and one of 13 in females) from VC-treated Hras2 mice; no mutations in murine Ki-ras were found in these tumors. Point mutations at codons 12 and 61 of the murine Ki-ras gene were observed, however, in one of 10 and six of 10 lung tumors respectively, from VC-treated non-Tg mice. These findings indicate that Hras2 mice are highly sensitive to pulmonary neoplasms and splenic and lung hemangiosarcomas after treatment with VC. The molecular analyses suggest that point mutations of the transgene and the murine Ki-ras gene do not play a major role in VC induction of pulmonary neoplasms in these transgenic mice. PMID- 9397191 TI - Induction of mouse cytochrome P450 2B enzymes by amine metabolites of musk xylene: contribution of microsomal enzyme induction to the hepatocarcinogenicity of musk xylene. AB - Musk xylene (MX) is a synthetic nitromusk perfume ingredient that, although uniformly negative in genotoxicity testing, causes liver tumors in B6C3F1 mice. MX is also capable of inducing cytochrome P450 enzymes in a manner similar to that of phenobarbital (PB), which suggests that epigenetic mechanisms may be involved in the carcinogenic response. At the same time, MX is metabolized in vivo by nitroreduction, a reaction catalyzed by intestinal flora that yields aromatic amine metabolites. These amine metabolites are also capable of inactivating CYP2B10, the major cytochrome P450 enzyme induced by MX treatment. In the study reported here, the monoamine metabolites of MX, o- and p-NH2-MX, were evaluated for their potential to induce CYP2B10 and CYP1A2 mRNAs. Northern blot analyses indicated that both amines markedly induced CYP2B10 mRNA, whereas CYP1A2 mRNA, the enzyme implicated in the bioactivation of aromatic amines and frequently induced by aromatic amines, was induced only slightly, a response that was not different from that seen with PB. Induction of CYP2B10 mRNA suggested that the amine metabolites may contribute to the enzyme induction profile seen with MX treatment. To test this hypothesis, mice were treated with broad-spectrum antibiotics (neomycin, tetracycline, and bacitracin) to eliminate the intestinal flora and prevent formation of o- and p-NH2-MX. In antibiotic-treated mice treated with MX (200 mg/kg) for 4 d, no evidence of microsomal enzyme induction was observed, including no increases in liver weight, total cytochrome P450 content, or CYP2B protein levels. These results indicate that the amine metabolites of MX are responsible for the enzyme induction seen after MX administration. Thus, the biochemical and molecular effects of amine metabolites of MX are markedly different from those of other aromatic amines but very similar to those of PB. Therefore, it appears that MX is a non-genotoxic chemical that may cause mouse liver tumors in a manner analogous to that of PB. PMID- 9397192 TI - Reverse transcription-polymerase chain reaction-based methodology to quantify differential gene expression directly from microdissected regions of frozen tissue sections. AB - Quantitative differences in the expression of oncogenes are a critical feature of the cancer process. Several methods are currently available for assessing differential gene expression, but none can be used to determine quantitative changes in gene expression from small numbers of cells. The ability to conduct this type of quantitative analysis would be useful in the study of definable, early stages of carcinogenesis when very few cells are involved. We therefore developed a highly sensitive, slide-based technique that incorporates the benefits of in situ polymerase chain reaction (PCR) and reverse transcription-PCR (RT-PCR) to quantify differential c-myc gene expression from liver tissue sections having either low or high levels of proliferating hepatocytes. To eliminate the need for isolating and quantifying mRNA, cells of interest were microdissected from frozen histological sections and their RNA directly subjected to RT-PCR amplification. These reactions were conducted in the presence of an internal RNA standard that was specifically designed to normalize differential RT and PCR efficiencies between samples. GENESCAN software analysis was used to determine the ratios of the RT-PCR products of the target gene to the RNA standard. These ratios were then normalized to the numbers of cells isolated, as quantified by image analysis, and comparative gene expression values were determined between sample groups. We conclude that this technology can be adapted to study any gene of interest in any type of frozen tissue or isolated cells. This methodology is particularly applicable to the molecular analysis of histopathologically distinct preneoplastic and neoplastic lesions identified on tissue sections. PMID- 9397194 TI - MHC class I and II expression in prostate carcinoma and modulation by interferon alpha and -gamma. AB - BACKGROUND: Expression of Major Histocompatibility Complex (MHC) class I and II antigens are critical for the cellular immune response. Loss of MHC expression represents one mechanism by which cancer cells escape immune recognition. PURPOSE: To define MHC class I and II expression by prostate cancer (PCa) in vivo and in vitro and the ability to modulate MHC expression in vitro with IFN-alpha and -gamma. METHODS: Frozen tissue sections of 25 benign prostatic hyperplasia (BPH) and 18 PCa specimens were studied by immunohistochemistry. PCa cell lines LNCaP, PC-3, and DU-145 were studied by FACS, ELISA, and cytospin. Class I was detected by monoclonal antibody (mAb) W6/32, and class II by mAb 13.17. The effects of IFN-alpha and -gamma were assessed by testing the three cell lines in the presence or absence of varying concentrations of the cytokine for varying incubation times. RESULTS: Class I was strongly expressed by 24/25 BPH specimens; 4/18 (22%) PCa were homogeneously class I-positive, while 5/18 (28%) were heterogeneously positive and 9/18 (50%) were class I-negative. PC-3 and DU-145 expressed normal levels of class I, while LNCaP expressed only low levels. All line except LNCaP demonstrated significant up-regulation of class I with either IFN-alpha or -gamma. Class II expression was not seen in BPH epithelium nor in 17/18 PCa. Class II could be only weakly induced in the three PCa lines. CONCLUSIONS: These findings confirm prior studies demonstrating that class I expression is commonly lost or diminished in PCa. In addition, class II up regulation by IFN-gamma appears very limited in relation to other normal or neoplastic epithelium. IMPLICATIONS: The present findings, taken together with previous studies, are most consistent with the expression of neoantigens by PCa, which are recognized and appropriately eliminated by the cellular immune system. This selective pressure favors outgrowth of cells which down-regulate or lose class I and/or class II expression. Understanding PCa immunobiology will help in the development of effective immunotherapy for this disease. PMID- 9397193 TI - Expression and localization of aminopeptidase A, aminopeptidase N, and dipeptidyl peptidase IV in benign and malignant human prostate tissue. AB - BACKGROUND: Cell-surface peptidases are ectoenzymes which regulate the access of bioactive peptides to their receptors on cell membranes. Abnormalities in their expression and function result in altered peptide activity which contribute to neoplastic transformation and/or progression. METHODS: Expression of aminopeptidase A (APA), aminopeptidase N (APN, CD13), and dipeptidyl peptidase IV (DPP IV, CD26) was immunohistochemically examined in 20 benign and 33 malignant prostate tissues (19 primaries and 14 metastases). RESULTS: Benign prostatic stroma exhibited no APA, APN, or DPP IV immunoreactivity. Stromal cells surrounding prostatic carcinoma cells demonstrated increased APA expression in 24/33 (73%) of tumors. Benign prostatic epithelial cells strongly expressed APN and DPP IV but not APA. In contrast, APN was expressed in > 80% of tumor cells in 5/33 (15%) of specimens, heterogeneously expressed (20-80% of cells positive) in 4/33 (12%) of specimens, and minimally expressed or absent in 24/33 (73%) of tumor specimens, with a similar pattern of expression in primary and metastatic tumors. DPP IV was expressed by > 80% of tumor cells in 18/19 (95%) of primary prostate cancer specimens, but in only 7/14 (50%) of metastases. CONCLUSIONS: These data show that cell-surface peptidases are differentially expressed by normal prostatic stromal and epithelial cells, with increased expression of APA in the stroma surrounding prostate cancer cells, absent APN expression in most tumor cells, and a decreased frequency of DPP IV expression in metastatic tumors. Further studies will elucidate the biological effects of the presence or loss of cell-surface peptidases in the benign and malignant prostate. PMID- 9397196 TI - Underutilization of health services among patients with urinary symptoms: results of a population-based survey in Israel. AB - BACKGROUND: Although there is firm evidence concerning the relatively-high rates of overutilization of prostate surgery among elderly men, only minimal efforts have been made to evaluate the existence and extent of underutilization. This assessment, accomplished by our study, may have a significant impact on health services planning and budgeting. METHODS: The study population comprised a nationwide representative sample of 960 Israeli men, aged between 45 and 75 years. Data were accumulated by personal interviews conducted at the homes of the individuals by trained staff. The questions included in the questionnaire aimed at describing the sociodemographic and clinical status. The responses to questions regarding male urinary symptoms were obtained by personal reports. RESULTS: Forty-three percent of the subjects reported having experienced urinary symptoms, but only 4.6% were severely bothered by the symptoms on a daily basis, and 8.9% were moderately bothered. Only 55.4% of patients with bothersome urinary symptoms visited their general practitioners, while only 39.7% were referred to a urologist because of their complaints. CONCLUSIONS: Elderly men bothered by urinary symptoms frequently do not seek health care. An educational program regarding the potential benefit of medical interventions for benign prostatic hypertrophy may significantly improve their quality of life. PMID- 9397195 TI - Heredity and prostate cancer: a study of World War II veteran twins. AB - BACKGROUND: Increased risk of prostate cancer among men with a family history of the disease has been observed in several epidemiological studies, and family studies have identified hereditary prostate cancer characterized by early onset and autosomal dominant inheritance. METHODS: In this study, we examine prostate cancer heritability among twins in the NAS-NRC Twin Registry, with cases ascertained from a number of sources: recent telephone interviews, Medicare and Department of Veterans Affairs hospitalizations, previous mail questionnaires, and death certificates. A total of 1,009 prostate cancer cases were identified among the cohort of 31,848 veteran twins born in the years 1917-1927. RESULTS: Probandwise concordance for prostate cancer was substantially higher among monozygous twin pairs, 27.1%, than among dizygous twin pairs, 7.1% (P < 0.001). CONCLUSIONS: These data suggest that genetic influences account for approximately 57%, and environmental influences for 43%, of the variability in twin liability for prostate cancer. PMID- 9397197 TI - Pamidronate administration improves the secondary hyperparathyroidism due to "Bone Hunger Syndrome" in a patient with osteoblastic metastases from prostate cancer. AB - BACKGROUND: The so-called Bone Hunger Syndrome is a metabolic derangement that sometimes complicates the natural history of prostate cancer patients with osteoblastic bone metastases. An excessive bone formation leads to calcium entrapment in bone and the subsequent increase of parathyroid hormone (PTH) levels, in response to calcium demand. PTH elevation stimulates the osteoclasts in sites distant from those involving the tumor, leading to osteomalacia. METHODS: PTH and markers of bone turnover were monitored every 3 weeks, from the start of pamidronate treatment in a prostate cancer patient with progressive disease, to luteinizing hormone releasing hormone analog (LHRH-A) administration, developing hyperparathyroidism, hypophosphatemia, and albumin corrected serum calcium close to the lower limit of normality. Serum bone alkaline phosphatase (BALP), assessed by two different methods: electrophoretic and immunoradiometric, and urinary levels of markers of bone collagen breakdown were also remarkably elevated. RESULTS: As a consequence of pamidronate infusion (60 mg e.v. every 3 weeks for a total of four times), BALP and PTH decreased consistently, serum calcium and phosphorus returned within the normal range, while markers of collagen resorption showed a significant decrease at the 9th week, preceded by a transient rise. CONCLUSIONS: This case report indicates that bisphosphonates could inhibit both osteoclast activity. The anti-osteoblastic effect is mainly responsible for the improvement of the pretreatment calcium imbalance of our patient towards hypocalcemia and the consequent hyperparathyroidism. PMID- 9397198 TI - Caloric restriction diminishes the age-associated loss of immunoreactive catalase in rat prostate. AB - BACKGROUND: Caloric restriction (CR) retards aging and diseases in mice, rats, and other animals by unknown mechanisms. A popular hypothesis is that CR acts by opposing age-associated increases in oxidative stress. METHODS: Because influences of CR on antioxidant enzymes in the prostate have not been previously investigated, immunohistologic methods (light and electron microscopy) were used to determine the prostatic localization of catalase (CAT) in rats of diverse ages (3-32 months) fed either normally or subjected to CR from age 16 months. RESULTS: In 20-month-old rats fed either diet, CAT appeared as dense deposits at the apical poles of the epithelium in the lateral lobes, and within the ductular lumens, suggesting that CAT is secreted. Confirmation of both liver peroxisomal and prostatic apical cytoplasmic localization of CAT was provided by electron microscopic immunogold staining. The amount of CAT was reduced at 30 months in normally fed rats but not in those on CR. CONCLUSIONS: CAT appears to be a secretory product of the epithelial cells in the lateral lobes of the rat prostate. Further, CR from late-middle age opposed the age-associated loss of this intracellular enzyme activity. PMID- 9397199 TI - Effects of Ca++ mobilization on expression of androgen-regulated genes: interference with androgen receptor-mediated transactivation by AP-I proteins. AB - BACKGROUND: The adult prostate is maintained through an equilibrium between cell growth and death rates. Androgen deprivation induces an increase in intracellular Ca++, AP-1 gene expression of androgen-inducible genes. METHODS: Northern blot analysis, band-shift assays, and transient cotransfection assays were used to study the effects of Ca++ mobilizer A23187 on gene expression in human prostate cancer cells. RESULTS: A23187 repressed androgen-upregulated mRNAs for prostate specific antigen (PSA) and hKLK2, and rapidly induced mRNA levels for c-fos and c jun. AP-1 protein-DNA binding activities were elevated after A23187 treatments. Androgen receptor (AR)-mediated induction of chloramphenicol acetyltransferase (CAT) reporter was repressed by AP-1 proteins. CONCLUSIONS: The repression of AR mediated induction of PSA and hKLK2 genes by Ca++ mobilizers is due to the interference of AR transactivation activity by AP-1 proteins. PMID- 9397200 TI - Localization of prostate cancer metastasis-suppressor activity on human chromosome 17. AB - BACKGROUND: Prostate cancer is the most commonly diagnosed malignancy in American men. Currently, it is difficult to accurately predict the clinical course of histologically localized prostatic cancer in the individual patient. Identification of markers for metastatic potential of prostate cancer may improve the diagnosis and treatment of this disease. We have previously demonstrated that human chromosome 17 (17pter-q23) suppresses the metastatic ability of AT6.1 rat prostatic cancer cells. In this study we report on the further localization of the metastasis suppressor activity encoded by human chromosome 17. METHODS: A series of AT6.1-17 microcell hybrids was constructed using microcell-mediated chromosomal transfer of human chromosome 17 into highly metastatic AT6.1 cells. Hybrids which had spontaneously deleted regions of chromosome 17 were analyzed by PCR for the presence of 32 sequence-tagged sites (STS) markers as well as the prostate cancer tumor-suppressor loci reported on 17q. In addition, we examined a number of candidate genes and markers that previously have been mapped to chromosome 17. The in vivo metastatic potential of these AT6.1-17 deletion hybrids was determined. RESULTS: We have localized metastasis-suppressor activity to a approximately 70-centiMorgan (cM) portion of chromosome 17, consisting of three distinct regions of 30 cM (D17S952-->D17S805), 6 cM (D17S930-->D17S797), and 34 cM (D17S944-->D17S784). Three of the four markers on 17p13, including HIC1 and TP53, and 12 of the 13 markers in 17q21-23, including BRCA1 (D17S855) and NM23 (NME1), were not retained in the conserved approximately 70-cM metastasis suppressor region. CONCLUSIONS: These results support a role for a novel metastasis-suppressor gene(s) or a novel metastasis-suppressor function on chromosome 17. Complementary candidate gene and positional cloning approaches are being used to identify the gene(s) within the approximately 70-cM conserved region responsible for metastasis suppression. PMID- 9397201 TI - Comparison of serum PSMA, PSA levels with results of Cytogen-356 ProstaScint scanning in prostatic cancer patients. AB - BACKGROUND: Stored serum from clinical trial cases undergoing ProstaScint (CYT 356) scanning were available for Prostate Specific Membrane Antigen (PSMA) assay. Prostate Specific Antigen (PSA) levels had already been determined. This provided an opportunity to see what correlations existed between the serum markers and the ProstaScint scan. A group of patients had the studies preprostatectomy, whereas another group had the studies postprostatectomy. METHODS: The scan results, serum PSA, serum PSMA, and clinical data were separately analyzed. PSMA serum levels were determined by Western blot. RESULTS: Preoperatively, radical prostatectomy patients showed a correlation between serum PSA or PSMA levels and the ProstaScint scan in the total group (n = 86), or in an untreated group (n = 38). Preoperatively, PSMA correlated with the pathological stage, whereas PSA correlated with the scan. Postoperatively, only PSMA serum levels correlated with the scan in an untreated group (n = 40). CONCLUSIONS: Preoperatively or postoperatively, Western blot PSMA serum levels predict the stage of disease or local, regional, or distant metastases, as shown by ProstaScint scan. Both the scan and the serum tests provide prognostic information and evaluate the extent of disease to a more significant degree than previously possible. PMID- 9397203 TI - Intrathoracic gastric volvulus from blunt abdominal trauma. A case report. PMID- 9397202 TI - HHV-8 (KSHV) does not establish latency in prostate cancer cell lines. AB - BACKGROUND: HHV-8 is a new herpesvirus found in lesions of Kaposi's sarcoma and some lymphoproliferative diseases. More recently, a report stated that normal prostate tissue also contains the virus. METHODS: The expression of HHV-8 was examined by a sensitive reverse-transcriptase PCR for the viral genes ORF 72, ORF 73, ORF 74, and ORF 75. In coculture experiments we attempted to infect 3 commonly studied prostate cancer cell lines using induced and uninduced lymphoid cell lines harboring HHV-8 (KS-1, BC-1, and BC-2). For induction of viral genes, butyrate and phorbol esters were used. RESULTS AND CONCLUSIONS: At baseline, prostate cancer cell lines LNCaP, DU-145, and PC-3 did not express viral gene products. Extensive coculture experiments were also negative. In no instance could latency for the virus be established. Our results argue against the involvement of HHV-8 in prostate cancer and for a limited tissue tropism of HHV 8. PMID- 9397204 TI - Polygalacturonase inhibitors in bean pods. AB - The amount of polygalacturonase-inhibiting protein (PGIP) was 14 times higher in bean pods than in etiolated hypocotyls. The PGIP was extracted from bean pods and partially purified by chromatography on columns of S-Sepharose. DEAE-Sephadex A 50, and Sephadex G-75. Further purification by ion-exchange chromatography on a Mono Q column separated two isoforms of the inhibitor. The two PGIPs were similar in most properties but differed slightly in pI values. They also differed in one residue of the N-terminal amino acid sequences. Both bean pod PGIPs differed in two and possibly three residues of the deduced N-terminal amino acid sequence for hypocotyl PGIP. Small alterations in the structure of PGIP may represent a strategy in bean plants for resistance to a variety of pathogens. PMID- 9397205 TI - Acyl secoiridoids and antifungal constituents from Gentiana macrophylla. AB - LC-UV-mass spectrometry and bioassay co-directed fractionation of an aqueous acetone extract of the roots of Gentiana macrophylla gave three new chromene derivatives and two novel and six known secoiridoids, along with kurarinone, kushenol I, beta-sitosterol, stigmasterol, daucosterol, beta-sitosterol-3-O gentiobioside, alpha-amyrin, oleanolic acid, isovitexin, gentiobiose and methyl 2 hydroxy-3-(1-beta-D-glucopyranosyl)oxybenzoate. The structures of the new products were established from spectral and chemical evidence as 2 methoxyanofinic acid and macrophyllosides A-D. The six known secoiridoids were gentiopicroside, sweroside, 6'-O-beta-D-glucosylgentiopicroside, 6'-O-beta-D glucosylsweroside, trifloroside and rindoside. The new acid (2-methoxyanofinic acid), its methyl ester, kurarinone and kushenol I were shown to be active against the plant pathogenic fungus Cladosporium cucumerinum. The methyl ester and kurarinone inhibited also the growth of the human pathogenic yeast Candida albicans. Structure-activity relationships were studied. Thus, addition of a methoxyl group to the benzene nucleus of anofinic acid (2,2-dimethyl-2H-1 benzopyran-6-carboxylic acid) increased the antifungal activity remarkably whereas glycosylation at the carboxylic moiety was found to remove the activity. Esterification of the new acid induced its activity against C. albicans, but decreased its growth inhibition properties against C. cucumerinum. Hydroxylation of kurarinone at the 3 beta-position removed its activity against C. albicans and decreased the inhibition of C. cucumerinum. In addition, the chemotaxonomic significance of the identified constituents is discussed. PMID- 9397206 TI - Antifungal and antibacterial naphthoquinones from Newbouldia laevis roots. AB - From a dichloromethane extract of Newbouldia laevis roots, four new (6 hydroxydehydroiso-alpha-lapachone, 7-hydroxydehydroiso-alpha-lapachone, 5,7 dihydroxydehydroiso-alpha-lapachone and 3-hydroxy-5-methoxydehydroiso-alpha lapachone) and six known naphthoquinones have been isolated. Their structures were established by spectroscopic methods (UV, EI mass spectrometry, 1H and 13C NMR) and that of 7-hydroxydehydroiso-alpha-lapachone was confirmed by X-ray crystallography. All naphthoquinones showed antifungal activity against Cladosporium cucumerinum and Candida albicans, and activity against the bacteria Bacillus subtilis and Escherichia coli. PMID- 9397207 TI - Steroid saponins from Solanum laxum. AB - Two new steroid saponins, named laxumins A and B, were isolated from the ethanolic extract of the aerial parts of Solanum laxum. These compounds were characterized, using mainly NMR spectroscopy, mass spectrometry and chemical methods, as (23S,25S)-spirost-5-en-3 beta, 15 alpha, 23-triol 3-O-{beta-D glucopyranosyl-(1-->2)- beta-D-glucopyranosyl-(1-->4)-[alpha-L-rhamnopyranosyl-(1 ->2)]-beta-D- galactopyranoside} and 3-O-{beta-D-glucopyranosyl-(1--> 4)-alpha-L rhamnopyranosyl-(1-->2)]-beta-D-galactopyranoside}, respectively. PMID- 9397208 TI - Steroidal saponins from fruits of Tribulus terrestris. AB - Further studies on the constituents of the fruits of Tribulus terrestris led to the isolation of five new steroidal saponins (terrestrosin A-E), (25R,S)-5 alpha spirostan-3 beta-ol-3 -O-beta-D-galactopyranosyl(1-2)-beta-D- glucopyranosyl(1-4) beta-D-galactopyranoside, (25R,S)-5 alpha-spirostan-3 beta-ol-3-O-beta-D glucopyranosyl(1-4)-[alpha-L- rhamnopyranosyl(1-2)]-beta-D-galactopyranoside, (25R,S)-5 alpha-spirostan-12-on-3 beta-ol-3-O-beta-D-galactopyranosyl (1-2)-beta D-glucopyranosyl(1-4)-beta-D-galactopyranoside, hecogenin 3-O-beta-D galactopyranosyl)1-2)-[beta-D- xylopyranosyl(1-3)]-beta-D-glucopyranosyl(1-4) beta-D-galactopyranoside and (25R,S)-5 alpha-spirostane-2 alpha, 3 beta-diol-3- O beta-D-galactopyranosyl(1-2)-beta-D-glucopyranosyl(1-4)-beta-D- galactopyranoside, together with five known steroidal saponins, desgalactotigonin, F-gitonin, desglucolanatigonin, gitonin and tigogenin 3-O-beta D- xylopyranosyl)1-2)-[beta-D-xylopyranosyl)1-3)]-beta-D-glucopyranosyl)1-4 )- [alpha-L-rhamnopyranosyl(1-2)]-beta-D-galactopyranoside. The structures of the new saponins were elucidated on the basis of spectroscopic analyses, including two-dimensional NMR techniques, and chemical reactions. PMID- 9397209 TI - Tiagabine approved for partial seizures. PMID- 9397210 TI - Antipsychotic choices now include quetiapine. PMID- 9397211 TI - Growth hormone-releasing hormone product cleared for marketing. PMID- 9397212 TI - NCQA data show wide variability in health plan performance. PMID- 9397213 TI - HMO patients receive less costly end-of-life care. PMID- 9397214 TI - APhA approves Guidelines for Pharmacy-Based Immunization Advocacy and Administration. PMID- 9397215 TI - Free vaccines, insurance status influence immunization by physicians. PMID- 9397217 TI - Benefits of a board-certified technician. PMID- 9397216 TI - New rules will standardize labeling for dietary supplements. PMID- 9397218 TI - Levofloxacin and trovafloxacin: the next generation of fluoroquinolones? AB - The pharmacology, spectrum of activity, pharmacokinetics, clinical efficacy, and adverse effects of levofloxacin, recently approved by FDA, and trovafloxacin, currently undergoing clinical trials, are reviewed. Compared with quinolones in current use, levofloxacin is more potent against gram-negative bacteria and exhibits better antipseudomonal activity as well as greater oral bioavailability. Trovafloxacin is more potent than existing quinolones against gram-positive bacteria. Both agents exert their antibacterial effects by inhibiting bacterial DNA synthesis. Compared with other quinolones, levofloxacin and trovafloxacin both demonstrate superior activity against the Bacteroides fragilis group, Chlamydia spp., Mycoplasma pneumoniae, and Mycobacterium spp. The half-life (t1/2) of levofloxacin is nearly eight hours. Levofloxacin can therefore be administered once daily for mild to moderate infections and twice daily for more serious infections. The recommended daily dose is 500 mg. Trovafloxacin has a t1/2 of 12 hours, which allows for single daily doses, and is extensively metabolized. Levofloxacin has demonstrated clinical efficacy in the treatment of community-acquired respiratory-tract infections, genitourinary infections, skin and skin-structure infections, acute bacterial sinusitis, and infections of the head and neck. Trovafloxacin may have a role in treating skin and skin-structure or soft-tissue infections respiratory-tract infections, sexually transmitted diseases, and meningitis. Both agents are well tolerated, with central-nervous system and gastrointestinal adverse effects reported most frequently. Concomitant administration of antacids or compounds containing meal cations decreases absorption of these quinolones. Levofloxacin and trovafloxacin have favorable antimicrobial and pharmacokinetic profiles, offering the advantages of once-daily doses as well as superior potency and spectrum of activity compared with currently available quinolones. PMID- 9397219 TI - Influence of patient information leaflets on anticonvulsant drug compliance in prison. AB - The effect of patient information leaflets on anticonvulsant drug compliance in a prison system was studied. Starting in January 1996, patient information leaflets were given to inmates in 18 prison units within the Texas Department of Criminal Justice who were receiving anticonvulsant medications. Nine of the units had pharmacist-operated chronic care clinics (POCs); the other nine did not. The units were matched on the basis of demographic characteristics and patient chronicity levels. Leaflet distribution continued until two weeks into March 1996. Certified medication aides (CMAs) watched inmates take doses and documented administration. A report on the anticonvulsant medications prescribed, dosage frequencies, number of patients taking each drug, and cumulative rate of compliance for each prison unit was generated monthly. The leaflets appeared to have a positive effect on compliance. The largest percent increases in compliance occurred on POC units, possibly because of the reinforcement provided by pharmacist counseling. The findings may have been affected by the integrity of the CMAs, the possibility that some patients on POCs did not receive pharmacist counseling, exploitation of the system by inmates for secondary gain, and other factors. Patient information leaflets may be of some use in increasing drug compliance in the prison population, but expectations must be tempered by the realities of this setting. PMID- 9397220 TI - Stability of thiotepa (lyophilized) in 0.9% sodium chloride injection. AB - The stability of thiotepa in a new formulation of the drug was studied. Vials of Thioplex (Immunex), a relatively new lyophilized formulation of thiotepa, were reconstituted with sterile water and diluted with 0.9% sodium chloride injection in polyvinyl chloride infusion bags to thiotepa concentrations of 0.5, 1, and 3 mg/mL. The solutions were stored at 8 and 25 degrees C in ambient light and analyzed at 0, 8, 24, and in most cases 48 hours for thiotepa concentration and chloro-adduct formation by stability-indicating high-performance liquid chromatography. Thiotepa 1 and 3 mg/mL was stable for 48 hours at 8 degrees C and for 24 hours at 25 degrees C. Thiotepa 0.5 mg/mL was not stable at either temperature. Storage at 8 degrees C slowed but did not prevent chloro-adduct formation and loss of potency. The pH tended to increase with time; turbidity remained low. Thiotepa (lyophilized) 1 and 3 mg/mL in 0.9% sodium chloride injection was stable for 48 hours at 8 degrees C and for 24 hours at 25 degrees C; the drug was unstable when diluted to 0.5 mg/mL and stored under the same conditions. PMID- 9397221 TI - Evaluation of electronic drug information resources for answering questions received by decentralized pharmacists. PMID- 9397222 TI - References for health-related quality-of-life claims in prescription drug advertisements. PMID- 9397223 TI - Visual compatibility of warfarin sodium injection with selected medications and solutions. PMID- 9397224 TI - Data sources for pharmacoeconomic and health services research. AB - Different types of databases available for health-related research, the data contained in these databases, and potential applications for pharmacists or researchers are discussed. Case studies that demonstrate uses for health databases are presented. Databases can be organized by facility, by health care provider, by disease or organ, or by sector. The types of data they contain include financial data, utilization data, demographic data, and outcomes data. Data can be obtained from the public sector, the private sector, or the researcher's own health system. The costs and time associated with using existing databases are often less than those required to collect data, but the quality and accessibility of the data must also be considered. The researcher's choice of database will depend on the research question. Health care databases can be used for health management and decision-making, quality review and evaluation, outcomes research, episode-of-illness studies, and evaluation of treatment protocols. Researchers must comply with patient-confidentiality and other agreements when accessing data. The format of the data needs to be matched with the hardware and software to be used in the analysis, and the data need to be loaded, verified, and cleaned before use. In deciding which of the many available data sources to use, researchers must determine the appropriate balance between external data and data available within their own health systems. The decision on whether to use existing data sources or to collect data prospectively will depend on the research question, the available resources, and the scope of the study. PMID- 9397225 TI - Gastrointestinal prokinetic agents for enhancing drug response in gastroparesis. PMID- 9397226 TI - Epoprostenol in the treatment of congestive heart failure. PMID- 9397227 TI - Anticonvulsant cross-sensitivity. PMID- 9397228 TI - Somatropin for treating AIDS-related wasting syndrome. PMID- 9397230 TI - The pharmacist's perspective on newer thrombolytic therapies for acute myocardial infarction.Introduction. PMID- 9397229 TI - Two cases of possible cefepime-induced neutropenia. PMID- 9397231 TI - Decision analysis in the formulary process. AB - The use of decision analysis as a tool in making formulary decisions is discussed. Decision analysis is best applied in formulary decisions when factors other than acquisition costs are important in determining overall treatment costs for two products. The decision-analysis process assigns probabilities and costs to various treatments and outcomes. In the case of acute myocardial infarction, the decision analyst would gather data on angioplasty and thrombolysis and assign probabilities and costs for each treatment and subsequent endpoints on the basis of clinical trial data. When such data do not exist, estimates may be generated by expert panels. Applying clinical trial data to an individual hospital is not straightforward because of differences between clinical trials and clinical practice. Analysts and clinicians should evaluate any proposed model for its robustness and adaptability to local conditions and practitioner variation. Access to internal hospital data is essential in developing the model. An ideal decision-analysis model includes all important available interventions and defines and discloses the analyst's time frame and financial perspective. After implementation of the formulary decision, the results can be monitored and, if necessary, adjustments can be made in the allocation of resources. Barriers to effective decision analysis include lack of data and differences in sources of cost and outcome data. Despite the current limitations of decision analysis, clinicians and policymakers may find this technique increasingly useful in the complex formulary process. PMID- 9397232 TI - Strategies for the management of acute myocardial infarction: selecting patients for thrombolytic therapy. AB - Strategies for managing acute myocardial infarction (AMI), with a focus on thrombolytics, are reviewed. Revised guidelines published by the American College of Cardiology and the American Heart Association strongly recommend the use of thrombolytic therapy in carefully selected patients to promote reperfusion of ischemic myocardium. Thrombolytics reduce in-hospital mortality, and the mortality benefit is maintained for at least one year. Which patients are the best candidates for thrombolytics has been debated; variables that have been analyzed include infarct location, time after onset of symptoms, age, sex, blood pressure, and prior AMI. Clinicians should be thoroughly familiar with the absolute and relative contraindications to thrombolytic therapy to minimize potential hemorrhagic complications. The diagnosis of AMI should be clearly established. All patients should receive thrombolytic therapy if they arrive for treatment within 12 hours of the onset of symptoms of AMI and have appropriate ECG changes. Aspirin should be given to all patients, and beta-blockers should also be given if there are no contraindications. Heparin may be given as antithrombotic therapy in patients not receiving thrombolytics or as adjuvant therapy in those receiving thrombolytics. Other adjuvant treatments, notably angiotensin-converting-enzyme inhibitors, are used as indicated. Primary angioplasty may have a role in selected patients. Long-term interventions are intended to prevent recurrence of AMI. Thrombolytic therapy can substantially improve survival and function in patients with AMI, especially when it is given within six hours of the onset of symptoms. PMID- 9397233 TI - A fresh look at the molecular pharmacology of plasminogen activators: from theory to test tube to clinical outcomes. AB - The molecular pharmacology of plasminogen activators and its implications for thrombolytic therapy are discussed. The benefits of coronary thrombolysis were first demonstrated with intracoronary and i.v. streptokinase. Tissue plasminogen activator (t-PA) or recombinant t-PA (alteplase) proved to be superior to streptokinase with respect to speed of reperfusion and reperfusion efficacy. Since alteplase neither lessened the risk of bleeding found with streptokinase nor generated Thrombolysis in Myocardial Infarction (TIMI) grade 3 flow rates above about 50%, the quest for faster-acting, safer, and more effective thrombolytic agents has continued. The ideal agent would be highly efficient at converting plasminogen to plasmin, have an intermediate half-life, have a low affinity for fibrin, and be of reasonable cost. Genetic engineering of the wild type t-PA molecule resulted in reteplase, which has a longer half-life than alteplase and may be superior in terms of lytic activity, myocardial salvage, and survival. Also under investigation are TNK-t-PA and n-PA, which have longer half lives and, in animal models, seem to produce more rapid and complete thrombolysis, at less risk of intracranial bleeding, than alteplase. The risk of intracranial bleeding remains a problem with all thrombolytics; the risk versus the benefit will have to be assessed in large randomized trials. An understanding of the functions of various regions of the t-PA molecule has led to genetic engineering of new and promising plasminogen activators. PMID- 9397234 TI - Clinical trials in thrombolytic therapy, Part 1: Outcome markers that go beyond mortality reduction. AB - The use of outcome markers other than mortality reduction alone for evaluating thrombolytic agents in patients with acute myocardial infarction (AMI) is discussed. Mortality has been a primary endpoint in clinical trials evaluating thrombolytic agents for treatment of AMI. However, differences in mortality rates among thrombolytics are 1% or less and require tens of thousands of patients to detect. Broadening the endpoints studied will allow for more extensive data collection and more comprehensive cost-effectiveness analysis, enabling clinicians to make better decisions. The Global Utilization of Streptokinase and Tissue Plasminogen Activator for Occluded Coronary Arteries (GUSTO-1) trial measured not only mortality but issues related to the patency of the infarct related artery and complications. Other potentially important outcome markers after AMI are left ventricular function; markers of reperfusion, such as early resolution of ST-segment elevation; and resolution of chest pain. Available long term data show that the mortality benefit from alteplase is sustained over time and is correlated with enzymatically determined infarct size, left ventricular function, the number of diseased vessels, and Thrombolysis in Myocardial Infarction flow grade at the time of discharge from the hospital. Clinicians must also consider risk factors for stroke. Outcome measures other than mortality alone may help in determining which thrombolytic agent is most effective clinically and in financial decision-making without requiring large, expensive trials. PMID- 9397235 TI - Clinical trials in thrombolytic therapy, Part 2: The open-artery hypothesis and RAPID-1 and RAPID-2. AB - The open-artery hypothesis as supported by thrombolytic study results is discussed. The open-artery hypothesis states that survival after acute myocardial infarction (AMI) is maximized by achieving early and sustained patency of the infarct-related artery. However, two large multicenter trials did not detect any difference in mortality between patients given alteplase and patients given streptokinase, despite previous evidence that alteplase led to earlier recanalization of infarct-related arteries. The Global Utilization of Streptokinase and Tissue Plasminogen Activator for Occluded Coronary Arteries (GUSTO-1) trial suggested that early and complete patency is essential for short term survival after AMI. Subsequent observations indicated that an open infarct related artery at the time of hospital discharge is associated with improved long term survival. In the Reteplase Angiographic Phase II International Dose-Finding (RAPID-1) trial, complete patency was more frequent in patients who received a double-bolus regimen of reteplase than in patients who received standard-dose alteplase. Similar results were obtained in the Reteplase versus Alteplase Patency Investigation during Myocardial Infarction (RAPID-2) trial, which compared the same double-bolus reteplase regimen with an accelerated regimen of alteplase. In both RAPID studies, mortality was lower and other outcomes were more favorable in reteplase recipients. Reteplase seems more likely to produce normal blood flow soon after AMI than either standard-dose or accelerated alteplase and may be associated with a lower mortality rate. This lends further support to the open-artery hypothesis. PMID- 9397236 TI - A prospective comparison of four study designs used in assessing safety and effectiveness of drug therapy in hypertension management. AB - The objective of the study was to compare prospectively the impact of study design on drug therapy safety and effectiveness data obtained in hypertension management. The main study was a randomized controlled clinical trial of four different prospective study designs used in postmarketing assessment involving 1008 primary care practices in nine Canadian provinces. Two thousand nine hundred sixty-four patients with mild to moderate hypertension received an angiotensin converting enzyme (ACE) inhibitor daily for 14 weeks in one of four postmarketing studies--a randomized double-blind clinical trial (RCT) (10 to 40 mg fosinopril daily v 5 to 20 mg enalapril daily), two structured open label trials of 10 to 40 mg fosinopril daily (one with free drugs), or an unstructured open label trial of 10 to 40 mg fosinopril daily. Patient demographic and baseline characteristics, systolic and diastolic blood pressures, adverse events reported, and data quality were recorded as the outcome measures. The results showed that the RCT patients were titrated to higher doses of ACE inhibitor than patients in the open studies, P < .008; patients in the open studies were more likely to receive adjuvant diuretic therapy, P < .008. The decrease in blood pressure was similar for patients in all four studies, mean decrease in systolic BP was between 18 and 20 mm Hg, mean decrease in diastolic BP was between 11 and 13 mm Hg. Fewer patients in the unstructured open trial reported adverse events than patients in the RCT; a 55% relative reduction in reported adverse events (P < .008) was associated with the unstructured trial. There were also fewer drug-related adverse events per patient reported in the unstructured study (17 per 100 patients) than in the other studies (27 to 41 per 100 patients), P < .008. Physician preference for rounding off blood pressure measurements to 0 or 5 occurred most often in the unstructured open trial (P < .008). In conclusion, despite differences in dose titration and in the use of adjuvant therapy, antihypertensive drug therapy effectiveness observed in an RCT may be similar to uncontrolled postmarketing studies. Open trials with scheduled follow-up visits are as effective in detecting severe adverse events as RCT, but postmarketing studies with unstructured schedules of follow-up are insufficient in identifying drug-related adverse events and have poorer quality data. PMID- 9397237 TI - Relation between nocturnal decline in blood pressure and mortality. The Ohasama Study. AB - To investigate the relation between nocturnal decline in blood pressure and mortality, we obtained ambulatory blood pressures in 1542 residents aged 40 years or over of a rural Japanese community. Subjects were followed-up for a mean of 5.1 years and were then subdivided into four groups according to the percent decline in nocturnal blood pressure: 1) extreme dippers: percent decline in nocturnal blood pressure > or = 20% of the daytime blood pressure; 2) dippers: decline of > or = 10% but < 20%; 3) nondippers: decline of > or = 0% but < 10%; and 4) inverted dippers: no decline. The relationship between the decline in nocturnal blood pressure and mortality was examined by the Cox proportional hazards regression model adjusted for age, sex, smoking status, previous history of cardiovascular disease, and the use of antihypertensive medication. The mortality risk was highest in inverted dippers, followed by nondippers. There was no difference in mortality between extreme dippers and dippers. This relationship was observed for both treated and untreated subjects, was more pronounced for cardiovascular than for noncardiovascular mortality, and did not change after the data were adjusted for 24-h, daytime, and nighttime blood pressure levels. PMID- 9397238 TI - Effects of N omega-nitro-L-arginine and N-acetyl-L-cysteine on the reversal of one-kidney, one-clip hypertension. AB - The present study evaluated whether nitric oxide (NO) synthesis blockade or potentiation (with N omega-nitro-L-arginine or N-acetyl-L-cysteine, respectively) modulates the systemic and renal responses to unclipping in anesthetized one kidney, one-clip hypertensive rats (1K-1C). Cardiac output was measured by thermodilution. In time-control rats, mean arterial pressure (MAP) decreased from 197 +/- 8 mm Hg to 139 +/- 4 mm Hg 3 h after unclipping, and cardiac index (CI) decreased by 35%, with a transient rise in sodium and water excretion and no changes in total peripheral resistance (TPR), glomerular filtration rate (GFR), or renal plasma flow (RPF). Administration of N omega-nitro-L-arginine methyl ester (NAME, 10 micrograms/kg/ min) blunted the hypotensive (from 190 +/- 6 mm Hg to 157 +/- 3 mm Hg), diuretic and natriuretic responses and potentiated the decrease in CI (40%) observed after unclipping, whereas TPR increased by 103%. Also, in rats given NAME, GFR and RPF decreased by 20% and 45%, respectively, at the end of the experiment. The effect of N-acetyl-L-cysteine (NAC, 300 mg/kg), a sulfhydryl group donor that may protect NO from free radical destruction by forming an S-nitrosothiol compound, was also evaluated. NAC potentiated the depressor response to unclipping (from 180 +/- 5 mm Hg to 97 +/- 3 mm Hg), and GFR and RPF increased by 80% and 35%, respectively. These effects of NAC appear to be NO dependent, as they were blocked by simultaneous administration of NAME. However, no significant differences were observed among groups in cumulative excretion of sodium and water, demonstrating that the hemodynamic effects of NAME and NAC after unclipping are due to mechanisms other than renal excretory changes. The results of the present study indicate that the cardiovascular depressor effects of unclipping are modulated by endothelium-derived nitric oxide. PMID- 9397239 TI - Synergistic effect of angiotensin II and a high sodium diet on the vascular glycosaminoglycan synthesis of rats. AB - The effect of dietary sodium supplementation on angiotensin II (ANG II)-induced stimulation of vascular glycosaminoglycan (GAG) synthesis of rats was investigated. Measurements were performed both ex vivo and in vivo to validate the in vivo measurements and to estimate the relative contribution of intrinsic and extrinsic influences to ANG II-stimulated GAG synthesis in sodium-fed rats. Male Sprague-Dawley rats on normal (0.7% NaCl) and high sodium (4.0% NaCl) diets were infused with 100 ng/kg/min ANG II intraperitoneally for 48 h, or sham operated (controls). To measure tissue GAG synthesis, 35SO4 was added to the incubation medium (ex vivo) or injected intravenously into rats (in vivo). Systolic blood pressure of ANG II-treated rats on normal sodium diet was unchanged, but it increased by 13 mm Hg (P < .05) in the rats on the combined treatment of ANG II and a high sodium diet. ANG II or high sodium diet by itself had no effect on GAG synthesis. On the combined treatment, GAG synthesis of the aorta was increased by 17% (P < .05) when measured ex vivo, and by 38% (P < .01) when measured in vivo. In vivo, there was also increased GAG synthesis of mesenteric arteries (P < .01) and of vena cava (P < .02); GAG synthesis of nonvascular tissue (diaphragm, bladder, and kidney) was unchanged. The synergistic effect of ANG II treatment and high sodium diet on GAG synthesis appears to be, in part, arterial pressure independent and vascular tissue specific. The greater effect of combined treatment in vivo than ex vivo suggests that humoral or neural stimuli may be contributing to the interaction. PMID- 9397240 TI - Glucose metabolism and insulin receptor binding and mRNA levels in tissues of Dahl hypertensive rats. AB - Increased insulinemic response to an oral glucose load has been demonstrated in Dahl salt-sensitive hypertensive rats. To determine whether this abnormality is mediated at the level of the insulin receptor, we compared insulin receptor binding and mRNA levels in tissues of Dahl salt-sensitive rats (DS) and in their normotensive controls, Dahl salt-resistant rats (DR). To evaluate possible influences of dietary sodium intake, rats were fed either low (0.07% NaCl) or high salt (7.5% NaCl) chow until the DS became hypertensive, and then were killed by decapitation. Fasting plasma glucose and plasma insulin levels did not differ between DR and DS rats and were not affected by salt intake. In response to an oral glucose load, plasma glucose had a similar increase in DR and DS rats, but the increase in plasma insulin was significantly greater in DS rats. Scatchard analysis of binding was obtained from in situ autoradiographic studies performed in frozen skeletal muscle and kidney sections, and insulin receptor mRNA levels were measured by slot-blot hybridization. Number and affinity of insulin receptors were comparable in skeletal muscle and kidney of DR and DS rats and, in both groups, binding parameters were not affected by dietary sodium chloride. Hepatic and renal insulin receptor mRNA levels were also comparable in DR and DS rats fed either low or high salt chow. Thus, increased plasma insulin response to oral glucose load is associated with normal insulin receptor binding and gene expression in peripheral tissues in rats with Dahl hypertension. A postreceptor defect is likely responsible for the decreased sensitivity to insulin in this model of genetic hypertension. PMID- 9397241 TI - Augmented contributions of voltage-gated Ca2+ channels to contractile responses in spontaneously hypertensive rat mesenteric arteries. AB - The observation that organic Ca2+ channel blockers are more effective in lowering blood pressure and peripheral resistance in hypertensive compared to normotensive subjects suggests that there is a greater contribution from voltage-gated Ca2+ channels (CaL) to vascular force maintenance in hypertensive arteries. This study tests this hypothesis by comparing the effects of Bay k 8644 and nisoldipine on basal force development, contractile responses to norepinephrine and serotonin, and Ca2+ currents (ICa) in mesenteric artery (MA) from Wistar-Kyoto rats (WKY) and spontaneously hypertensive rats (SHR). MA rings were used to record isometric contractions at Lmax. Single cells were isolated by collagenase plus elastase for measurement of CaL properties by patch-clamp methods. Contractile responses to Bay k 8644 were larger and more sensitive in SHR than WKY, and were larger in endothelium-denuded compared to intact rings. In SHR, the addition of 10 nmol/L Bay k 8644 increased contractile sensitivity to norepinephrine (NE) and serotonin (5HT), and increased maximum response to 5HT. In WKY, 10 nmol/L Bay k 8644 produced a small increase in 5HT sensitivity with no effect on maximum response, and had no effect on NE responses. In the presence of 1 mumol/L nisoldipine, the maximum response and the sensitivity to both NE and 5HT were decreased in both WKY and SHR with the inhibitory effects of nisoldipine being larger in SHR than WKY. Peak ICa was larger in SHR, and current-voltage curves were shifted toward more negative voltages compared to WKY. Bay k 8644 increased ICa in both WKY and SHR myocytes with no apparent difference in the magnitude of its effect when expressed as a percent of control ICa. These results suggest that CaL contribute significantly to tonic force maintenance as well as to agonist responses in MA from both WKY and SHR, but with a much larger contribution in SHR. Differences in the sensitivity of CaL to Bay k 8644 were not responsible for the differences in contractile responses to this agonist. PMID- 9397242 TI - The role of vasopressin in essential hypertension. Plasma levels and effects of the V1 receptor antagonist OPC-21268 during different dietary sodium intakes. AB - To study the role of vasopressin (VP) in essential hypertension, we examined plasma levels of VP and blood pressure (BP) response to an orally active V1 receptor antagonist, OPC-21268, in hypertensive patients on diets with different sodium contents. Plasma VP was determined in 12 normotensive subjects and 12 patients with mild essential hypertension on a regular sodium diet, and in eight hypertensive patients on a high sodium (250 mmol/day) and a low sodium (25 mmol/day) diet. BP response was examined for 4 h after single oral administration of OPC-21268 (100 mg) or placebo in eight patients on the regular diet, and in six patients on the high and low sodium diets. In four patients on the regular diet, the effects of OPC-21268 on the baroreflex control of heart rate were also examined with intravenous injections of methoxamine. Plasma VP did not differ between the normotensive and hypertensive subjects. Levels of VP in the plasma was higher in the high sodium than in the low sodium period, but the difference was not significant. BP and heart rate did not change significantly after administration of OPC-21268 or placebo under either condition. OPC-21268 also failed to lower BP in salt-sensitive patients on the high sodium diet. The baroreceptor reflex sensitivity was not modified by the administration of OPC 21268. Our results suggest that VP does not play an important role in mild essential hypertension through its action on the V1 receptors regardless of dietary sodium intake. PMID- 9397243 TI - Insulin resistance is related to silent cerebral infarction in patients with essential hypertension. AB - Recently, hyperinsulinemia or insulin resistance has been suggested to be a risk factor for cardiovascular diseases. We evaluated the role of insulin resistance in the occurrence of silent cerebral infarction in 28 patients with essential hypertension (40 to 75 years, 157 +/- 4/89 +/- 2 mm Hg). Patients with diabetes mellitus or obesity (BMI > or = 30) were excluded. Insulin resistance was evaluated by means of constant glucose infusion rate (M value) during euglycemic hyperinsulinemic glucose clamp test. Infarction was defined as a focal area with prolonged T1 and T2 relaxation times that was > 5 mm in diameter on brain magnetic resonance imaging. The severity of periventricular hyperlucency was evaluated by the distribution of the high intensity area. The number of silent infarctions significantly correlated only with the M value (F = 7.58, R2 = 0.23, P = .01) in multiple regression analysis using all variables: age, blood pressure, smoking history, lipid profile, levels of plasma glucose and insulin on fasting, and total amounts during 75-g OGTT. However, the severity of periventricular hyperlucency did not show a correlation with any factors. The occurrence of cerebral infarction was significantly correlated with thickening of the intima-media complex (IMC) of the common carotid artery on B-mode ultrasonography (F = 8.43, R2 = 0.25, P < .01). In conclusion, insulin resistance and thickening of IMC show a close relationship with the occurrence of silent cerebral infarction. Therefore, it may be important to improve insulin resistance for prevention of cerebral infarction in essential hypertensives. PMID- 9397244 TI - Myocardial wall thickness and left ventricular geometry in hypertensives. Relationship with insulin. AB - In hypertensive patients the presence of left ventricular (LV) hypertrophy has been associated with a more severe degree of insulin resistance. Whether myocardial wall thickness or LV geometry are associated with a different degree of insulin resistance is still unknown in essential hypertensives. For this reason 26 men with new diagnosed essential hypertension were enrolled. All patients underwent echocardiographic examination and euglycemic hyperinsulinemic glucose clamp combined with indirect calorimetry. According to LV mass and relative wall thickness data, all patients were categorized in four groups: 1) patients with a normal geometric LV pattern (n = 8) (PAT = 0); 2) patients with concentric remodeling LV mass (n = 8) (PAT = 1); 3) patients with eccentric LV hypertrophy (n = 3) (PAT = 2); and 4) patients with concentric LV hypertrophy (n = 7) (PAT = 3). All groups were similar for anthropometric characteristics. Patients with normal echocardiographic LV pattern (PAT = 0) had higher whole body glucose disposal (WBGD), oxidative and nonoxidative glucose metabolism, and lower lipid oxidation than patients with abnormal echocardiographic LV patterns (PAT = 1 to 3). Nevertheless, no significant differences among the groups with abnormal echocardiographic patterns were found. After controlling for age, body mass index (BMI), waist/hip ratio (WHR), and mean arterial blood pressure, only sum of the wall thickness was significantly correlated with fasting plasma insulin (r = 0.38, P < .05), WBGD (r = - 0.50, P < .009), and NOGM (r = - 0.48, P < .02). In multivariate analysis, a model made by age, BMI, WHR, systolic and diastolic blood pressure, and WBGD explained 38% of the echocardiographic pattern variability. In this model, WBGD (P < .02) was significantly and independently associated with echocardiographic patterns explaining 19% of the echocardiographic pattern variability. In conclusion, our data demonstrate that in arterial hypertension hyperinsulinemia/insulin resistance mainly affects myocardial wall thickness, whereas only a trivial association with LV geometry occurs. PMID- 9397245 TI - Effect of doxazosin on endothelial dysfunction in hypercholesterolemic/antioxidant-deficient rats. AB - Hypertension, hypercholesterolemia, atherosclerosis, and coronary heart disease are associated with abnormal endothelium-dependent, nitric oxide-mediated vasorelaxation. In rats, hypercholesterolemia in combination with deficiencies of vitamin E and selenium results in increased endogenous lipid oxidation and endothelial dysfunction. Two hydroxymetabolites of doxazosin, an alpha 1 adrenergic blocking antihypertensive agent, inhibit human lipid oxidation in vitro in a dose-dependent fashion. The present studies were performed to determine the effect of in vivo treatment with doxazosin on endothelial dysfunction in hypercholesterolemic/ antioxidant-deficient rats. Dahl rats were fed 1) a standard diet, 2) a high cholesterol (4%) diet, or 3) a high cholesterol, vitamin E- and selenium-deficient diet. A subgroup of animals in each group were administered doxazosin (3.5 mg/100 g/day) for 16 weeks. In the aortas, vascular relaxations induced by acetylcholine were significantly decreased (P < .05) in high cholesterol/antioxidant-deficient rats compared with normal and high cholesterol animals. Doxazosin treatment prevented the impairment in endothelium-dependent vascular relaxation in the high cholesterol/antioxidant deficient group. Vasorelaxation in response to the exogenous nitric oxide donor diethylamine nanoate, which was significantly impaired (P < .05) in aortas from high cholesterol/antioxidant-deficient animals compared with normal and high cholesterol animals, was normalized in aortas from high cholesterol/ antioxidant deficient animals that had received doxazosin. The antioxidant effect of doxazosin may have therapeutic implications in diseases associated with endothelial dysfunction linked to products of lipid oxidation. PMID- 9397246 TI - Comparison of amlodipine and long-acting diltiazem in the treatment of mild or moderate hypertension. AB - The comparative effects of the once a day calcium channel antagonists amlodipine and long-acting diltiazem were assessed in a parallel design, investigator blinded, multicenter trial in 123 patients with diastolic blood pressures ranging from 95 to 114 mm Hg before treatment. Patients were randomized to one of the two drugs and titrated at 2-week intervals to 5 or 10 mg of amlodipine or 180, 240, or 360 mg of long-acting diltiazem during a 10-week treatment period. Both drugs significantly reduced resting, sitting, standing, and 24-h ambulatory systolic and diastolic pressures. Amlodipine caused significantly greater reductions in sitting and standing systolic pressures, standing diastolic pressures, and 24-h ambulatory systolic and diastolic pressures versus diltiazem. Sitting systolic pressures were reduced from 151.9 +/- 2.0 (SE) at baseline to 137.9 +/- 1.8 mm Hg with amlodipine treatment and from 149.0 +/- 2.1 to 145.1 +/- 2.5 mm Hg with diltiazem. Sitting diastolic pressures were reduced from 100.2 +/- 0.6 to 87.8 +/ 1.0 mm Hg with amlodipine and from 101.1 +/- 1.0 to 91.9 +/- 1.1 mm Hg with diltiazem. Reductions in standing systolic pressures after treatment were -12.1 +/- 1.5 mm Hg amlodipine v -4.6 +/- 1.5 mm Hg diltiazem (P < .01), and reductions in standing diastolic pressures were -11.8 +/- 0.9 mm Hg amlodipine v -8.6 +/- 0.9 mm Hg diltiazem (P < .02). Heart rates did not change significantly with either drug during the study. Two subjects in each group dropped out because of adverse experiences. Although both agents were well tolerated and reduced blood pressures consistently over the 10-week test period, amlodipine was more effective than diltiazem in reducing systolic and diastolic blood pressures to the target pressures of < 140 mm Hg systolic and < 90 mm Hg diastolic over a range of doses widely used in clinical practice. PMID- 9397247 TI - Home blood pressure as a predictor of future blood pressure stability in borderline hypertension. The Tecumseh Study. AB - We evaluated time-related blood pressure trends in the Tecumseh study participants, none of whom received antihypertensive treatment. At baseline the blood pressures were measured in the field clinic and by self measurement at home (twice daily for 7 days). After a mean of 3.2 +/- 0.42 years, the clinic and home pressure readings were repeated. Nine hundred forty-six subjects had clinic and home blood pressure readings at baseline. Of these 735 (380 men, 355 women; average age, 32 years) also completed the second examination. Blood pressure, morphometric data, and biochemical measures at the first examination were used as predictors of future clinic blood pressures. Five hundred ninety-six subjects were normotensive on both examinations (81%). Of 79 subjects (10.7%) with clinic hypertension (> 140 mg Hg systolic or 90 mm Hg diastolic) at baseline, 38 remained hypertensive ("sustained hypertension") and 41 became normotensive ("transient hypertension") after 3 years. Another 60 normotensives at baseline (10.4%) became hypertensive on second examination ("de novo hypertensives"; incidence; 8.1%). The home blood pressure readings on both examinations were reproducible. The three hypertensive groups had elevated home blood pressure, were overweight, had dyslipidemia, and higher insulin values. Only the home blood pressure proved predictive of subsequent blood pressure trends. A home blood pressure of 128 and 83 mm Hg or higher detected "sustained" hypertension with a 48% sensitivity and 93% specificity. Readings of 120 and 80 mm Hg or lower predicted future normotension with a 45% sensitivity and a 91% specificity. We conclude that self determination of the blood pressure at home is useful in the management of borderline hypertension. An algorithm for the management of these patients is proposed. PMID- 9397248 TI - Factors that affect blood pressure variability. A community-based study in Ohasama, Japan. AB - Factors that affect blood pressure (BP) variability, ie, standard deviation (SD) and variation coefficient (VC: SD/average ambulatory BP) of ambulatory BP, were examined in a community-based sample in northeastern Japan. Screening and ambulatory BPs were measured in 823 subjects > or = 20 years of age, and the effects of age and BP on the SD and the VC were examined. In bivariate regression analysis, the SD of ambulatory BP was positively correlated with age and the ambulatory BP. The VC was also correlated with age. Both the SD and the VC were strongly correlated with the magnitude of the nocturnal decline in BP. Ambulatory BP was positively correlated with age and negatively correlated with heart rate and the SD of heart rate. Multivariate analysis demonstrated that the nocturnal decline in BP showed the strongest association with the SD and the VC of 24-h BP. However, age and BP were still independently and positively associated with the SD and the VC of ambulatory BP. Furthermore, pulse pressure and BMI were independently and positively associated with the SD and the VC of ambulatory BP. Since the SD and the VC of 24-h ambulatory BP were determined mainly by the nocturnal decline in BP, this variable appears to be an index of the circadian variation in BP and not an index of short-term BP variability. Pulse pressure, an index of arterial stiffness, was a relatively strong predictor of the SD and the VC of BP. In addition, the SD of heart rate, an index of baroreflex function, decreased with increasing age. Findings suggest that the increase in BP variability in hypertensive and elderly subjects may be explained, in part, by a disturbance of baroreflex function associated with an increase in arterial stiffness due to aging and hypertension. PMID- 9397249 TI - Captopril test and renal duplex scanning for the primary screening of renovascular disease. AB - To evaluate the utility of renal duplex scanning and the captopril test in the detection and functional assessment of renovascular disease, by comparing their results with those of angiography and captopril isotopic renography (CIR). Sixty hypertensive patients with aortoiliac disease and 16 with clinically suspected renovascular hypertension (RVH) were included. All the patients underwent renal duplex scanning prior to angiography. In addition, isotopic renograms and a determination of peripheral plasma renin activity (PRA) at baseline and 60 min after oral intake of 50 mg of captopril were both performed. A postcaptopril PRA > 5.7 ng/mL/h was considered as diagnostic of a positive captopril test. On the basis of the results of the angiography and isotopic renograms, all the patients were classified into three groups: group I (n = 33), essential hypertension (EHT); group II (n = 20), hypertension and angiographic RAS > 60% but negative CIR; and group III (n = 24), RAS > 60% and positive CIR. This last condition was considered as highly suspicious for RVH. Renal duplex scanning showed greater accuracy than captopril PRA or CIR for detecting RAS > 60% (groups II and III) with 87.3% versus 52.4% and 45.3% sensitivity (S), and 91.5% versus 84.4% and 92.8% specificity (Sp), respectively. The captopril test correctly identified 44 of 51 EHT patients (groups I and II) and 20 of 23 highly suspected of RVH (group III) with 87% S, 86.5% Sp, 74.1% PPV, and 93.6% NPV. Accuracy was further increased when a combined approach (renal duplex scanning and captopril test) was followed (82.6% S, 93.7% Sp, 86.4 PPV, and 91.8 NPV). In our study, renal duplex scanning was a useful screening method for detecting anatomical RAS. A combination of both renal duplex scanning and captopril test may be an appropriate approach to the primary screening for RVH, thereby permitting the selection of those patients indicated for angiography. PMID- 9397250 TI - Pressure modulates monocyte migration. AB - Migration of monocytes into the subendothelial space of the aorta has been considered to be an important event in the development of atherosclerosis. Because hypertension is commonly associated with atherosclerosis, we studied the effect of applied pressure on the migration of monocytes. Direct applied pressure increased the migration (P < .001) of monocytes across a filter when compared with normal atmospheric pressure. The migration of monocytes was found to be directly related to the amount of the applied pressure. Amlodipine, a calcium channel blocker, attenuated the migration of monocytes under normal as well as increased pressure conditions in a dose-dependent manner. These studies provide a basis to speculate on the role of direct pressure in the migration of monocytes into the subendothelial space and the possibility that vasoactive agents may modulate the migration of monocytes independent of their pressure-lowering effect. PMID- 9397251 TI - White-coat resistant hypertension. AB - The aim of this study was to evaluate whether sustained hypertensives with high clinic blood pressure, despite multiple drug treatment, show a true resistant hypertension or a "white-coat effect," and whether the pretreatment white-coat effect is maintained despite pharmacological therapy. The occurrence of resistant hypertension was determined in 250 consecutive essential hypertensives who had had an ambulatory blood pressure monitoring before treatment assignment. Twenty seven of 250 hypertensives with persistently high clinic blood pressure despite 3 months of adequate pharmacological therapy underwent further ambulatory blood pressure monitoring. Using our internal standards, seven patients had a true resistant hypertension whereas 20 subjects showed a large white-coat effect (white-coat resistant hypertension), ie, high clinic blood pressure (> 140/90) but "normal" ambulatory daytime (< 139/90 mm Hg) and 24 h (135/85 mm Hg) blood pressure. Using other cutoff points for ambulatory blood pressure, 134/90 and 135/85 mm Hg for daytime blood pressure, 10 and 13 patients, respectively, were reclassified as true resistant hypertensives and 17 and 14, respectively, were white-coat resistant hypertensives. Interestingly, in white-coat resistant hypertensives the large differences between clinic and ambulatory daytime blood pressure (white-coat effect), recorded before treatment assignment, were not affected by drugs and remained constant over time. Left ventricular mass index in white-coat resistant hypertensives was significantly lower than in truly resistant hypertensives, suggesting that prognosis could differ between these groups. In this study, using either our internal standards or some other cutoffs reported in the literature, the white-coat phenomenon was an important cause of resistant hypertension. The use of ambulatory blood pressure monitoring in these patients may avoid misdiagnosis of resistant hypertension, unnecessary overtreatment, and expensive procedures to look for possible secondary hypertension. PMID- 9397252 TI - Association study between the ANF gene and hypertension in a Gulf Arab population. AB - We have studied an insertion/deletion (I/D) dimorphism located in the second intron of the human atrial natriuretic factor (ANF) gene among 232 United Arab Emirates (UAE) nationals (112 normotensives and 120 hypertensives) from the Abu Dhabi Emirate, with a view to evaluating the value of this marker in relation to hypertension. Our findings show that genotype frequencies of this I/D marker occur in Hardy-Weinberg proportions (respective genotype frequencies in the overall sample population are: II, 51%; ID, 42%; DD, 7%). No association, however, was evidenced between this dimorphic site and clinical diagnosis of essential hypertension. This suggests that: 1) this I/D dimorphism is not a useful marker to study the relationship between the ANF gene and hypertension in the UAE; and 2) variations of the ANF gene that may be in linkage disequilibrium with this marker do not play a major role in the determination of hypertension in this Arab population. PMID- 9397254 TI - Blood pressure measurement in a patient with thoracic outlet syndrome: a case report. PMID- 9397253 TI - Magnesium concentrations in plasma, erythrocytes, and platelets in hypertensive and normotensive obese patients. PMID- 9397255 TI - Do ambulatory blood pressure patterns of isolated systolic hypertension predict atherosclerotic complications? PMID- 9397256 TI - HpaII-polymorphism of the atrial-natriuretic-peptide gene and essential hypertension in whites. PMID- 9397257 TI - Subspecialization. PMID- 9397258 TI - Presidential address of the American Orthopaedic Society for Sports Medicine. Who are we? The past present, and future. PMID- 9397259 TI - A prospective study of prognostic factors concerning the outcome of arthroscopic surgery for anterior ankle impingement. AB - Sixty-two consecutive patients with painful limited dorsiflexion of the ankle not responding to nonoperative treatment participated in a prospective study. All 42 men and 20 women (average age, 31 years) underwent arthroscopic surgery. Preoperative radiographs were graded according to an osteoarthritic and an impingement classification. Standardized followup took place at 4 months and 1 and 2 years after surgery. Results showed that the degree of osteoarthritic changes is a better prognostic factor for the outcome of arthroscopic surgery for anterior ankle impingement than size and location of the spurs. The hypothesis is that osteophytes without joint space narrowing are not a manifestation of osteoarthritic changes but rather the result of local (micro)trauma. After 2 years, 73% of the patients experienced overall excellent or good results; 90% of those without joint space narrowing had good or excellent results, and 50% of those with joint space narrowing had good or excellent results. At the 2-year followup, the group without joint space narrowing showed significantly better scores in pain, swelling, ability to work, and engagement in sports. This study also revealed that patients with less than 2 years of ankle pain before surgery and spurs located anteromedially were more satisfied with the outcome than when longer periods of preoperative pain were involved and when spurs were located anterolaterally. PMID- 9397260 TI - Salvage surgery for lateral tennis elbow. AB - We undertook a retrospective analysis of 34 patients (35 elbows) who had prior failed surgical intervention for lateral tennis elbow. Revision surgeries were performed between 1979 and 1994. Each patient's non-operative and operative history was recorded before our salvage revision surgery. At revision surgery, findings included residual tendinosis of the extensor carpi radialis brevis tendon in 34 of 35 elbows. In 27 elbows, the pathologic changes in the extensor carpi radialis brevis tendon had not been previously addressed at all, and in 7 elbows the damaged tissue had not been completely excised. Salvage surgery included excision of pathologic tissue in the extensor carpi radialis brevis tendon origin combined with excision of excessive scar tissue and repair of the extensor aponeurosis when necessary. Based on a 40-point functional rating scale proposed here, 83% of the elbows (29 of 35) had good or excellent results at an average followup of 64 months (range, 17 months to 17 years). To prevent failure of surgical treatment for tennis elbow, the pathologic tissue usually present in the extensor carpi radialis brevis tendon should be resected. Release operations, which weaken the extensor aponeurosis but fail to address the pathoanatomic changes, are not recommended. PMID- 9397261 TI - The natural history of the anterior cruciate ligament-deficient knee. Changes in synovial fluid cytokine and keratan sulfate concentrations. AB - Restoring knee stability through reconstruction, while providing symptomatic relief, has not been shown to decrease the incidence of degenerative changes after rupture of the anterior cruciate ligament. This suggests that posttraumatic osteoarthritis may not be purely biomechanical in origin, but also biochemical. To test this, we measured the levels of seven cytokine modulators of cartilage metabolism in knee joint synovial fluid after anterior cruciate ligament rupture. We also measured keratan sulfate, a product of articular cartilage catabolism. The sample population consisted of patients with uninjured knee joints (N = 10), and patients with acute (N = 60), subacute (N = 18), and chronic (N = 8) anterior cruciate ligament-deficient knees. Synovial fluid samples were analyzed by enzyme linked immunosorbent assays. Normal synovial fluids contained high levels of the interleukin-1 receptor antagonist but low concentrations of other cytokines. Immediately after ligament rupture there were large increases in interleukins 6 and 8, tumor necrosis factor alpha, and keratan sulfate. Interleukin-1 levels remained low throughout the course. As the injury became subacute and then chronic, interleukin-6, tumor necrosis factor-alpha, and keratan sulfate levels fell but remained considerably elevated 3 months after injury. Concentrations of interleukin-1Ra fell dramatically. Granulocyte-macrophage colony-stimulating factor concentrations were normal acutely and subacutely but by 3 months after injury were elevated 10-fold. Our data reveal a persistent and evolving disturbance in cytokine and keratan sulfate profiles within the anterior cruciate ligament-deficient knee, suggesting an important biochemical dimension to the development of osteoarthritis there. PMID- 9397262 TI - The effects of hyaluronan on the meniscus and on the articular cartilage after partial meniscectomy. AB - The effect of hyaluronan (molecular weight = 8 x 10(5)) on the meniscus and on the articular cartilage was assessed after partial meniscectomy in a rabbit model. On gross examination, remodeled meniscus appeared as newly synthesized translucent tissue, and was seen in both vehicle- and hyaluronan-treated menisci. Histologically, safranin O staining revealed the strong presence of glycosaminoglycans in the newly remodeled tissue, and polarized light demonstrated the absence of mature collagen architecture. Hydration of the hyaluronan-treated menisci was significantly less than that of the vehicle treated menisci, and the reducible collagen cross-link dihydroxylysinonorleucine was significantly increased in the hyaluronan-treated menisci compared with the vehicle-treated menisci, indicative of a greater degree of collagen remodeling. In situ hybridization of vehicle- and hyaluronan-treated menisci revealed a high level of type I procollagen mRNA expression and minor expressions of types II and III mRNA. Expression of the type I collagen gene appeared to be more pronounced in the hyaluronan-treated menisci than in the vehicle-treated menisci. The tibial plateaus revealed mild cartilage fibrillation after partial meniscectomy. A statistically significant difference between vehicle- and hyaluronan-treated cartilage was not demonstrated in the present study because of the slow development (i.e., 12 weeks) of osteoarthritis after partial meniscectomy in the rabbit model. These results suggest that in the rabbit model, hyaluronan enhances collagen remodeling and inhibits meniscal swelling after partial meniscectomy in the avascular region. PMID- 9397263 TI - Distal radial growth plate injury and positive ulnar variance in nonelite gymnasts. AB - To assess the prevalence of stress injury to the distal radial growth plate and of positive ulnar variance in a nonelite gymnast population, we administered a radiographic survey and questionnaire to 44 skeletally immature nonelite gymnasts (27 girls and 17 boys). The subjects trained an average of 11.9 hours per week. Radiographic findings consistent with stress injury of the distal radial physis were found in 25% (11 of 44) of participants. Ulnar variance was found to be more positive in the gymnasts when compared with age-predicted norms. An average side to-side difference in ulnar variance of 0.9 mm was observed. Radiographic findings of stress injury to the growth plate and the amount of ulnar variance were not associated with age, sex, training intensity, wrist pain, height, or weight. There was also no significant relationship between ulnar variance and radiographic findings. The mean ulnar variance in nonelite gymnasts was between that measured for elite gymnasts and nongymnasts. These results indicate that stress injury of the distal radial growth plate occurs in a significant percentage of nonelite gymnasts. It also appears that ulnar variance is more positive than would otherwise be predicted, suggesting growth inhibition of the distal radius, a growth stimulation of the ulna, or a combination of both. PMID- 9397264 TI - Reconstruction of the anterior and posterior cruciate ligaments after knee dislocation. Use of early protected postoperative motion to decrease arthrofibrosis. AB - We report a critical rating of results for 11 patients with bicruciate ligament reconstructions and immediate protected knee motion after knee dislocations (seven acute and four chronic). Six patients had concurrent repair or reconstruction of medial ligamentous structures, and six had reconstruction of the lateral and posterolateral ligaments. All patients returned for followup at a mean of 4.8 years postoperatively. Follow-up arthrometric testing at 20 degrees of flexion showed 10 knees had less than 3 mm of increased total anterior posterior displacement and 1 knee had 7 mm of increase. At 70 degrees of flexion, 9 knees had less than 3 mm of increased displacement and 2 knees had more than 6 mm of increase. The failure rates were as follows: 18% of posterior cruciate ligament reconstructions (2 of 11), 9% of anterior cruciate ligament reconstructions (1 of 11), 17% of lateral and posterolateral procedures, and 0% of medial collateral ligament procedures. At followup, five of the seven patients with acute injuries had no limitations with daily or sports activities. Three of the four patients with chronic ruptures were asymptomatic with daily activities, but only one was asymptomatic with light sports. Five patients (all acute injuries) required treatment for knee motion limitations. Nine patients had full range of motion at followup. We concluded that simultaneous bicruciate ligament reconstructions, performed with associated medial or lateral procedures, are warranted to restore function to all ligament structures. Even though immediate motion was used, several patients required early manipulation or arthroscopic debridement, which restored full motion and prevented permanent arthrofibrosis. PMID- 9397265 TI - The reharvested central third of the patellar tendon. A histologic and biomechanical analysis. AB - We assessed the histologic, mechanical, and structural properties of the reharvested central-third patellar tendon in greyhounds. Twelve dogs had the central third of the patellar tendon (5 mm) removed with corresponding bone blocks from the patella and tibia; the remaining tendon defect was loosely closed. Six dogs were sacrificed at 6 months and six at 12 months, and the central third of the patellar tendon was harvested from both the operative and the contralateral control knees. Analysis of the structural changes in the tendons revealed a significant increase in thickness for reharvested tendons at both 6 and 12 months when compared with controls. The entire residual tendons were narrower at 6 months and were shorter at 12 months compared with controls. Mechanical testing showed that the average failure load, ultimate tensile strength, strain at failure, and average modulus for the reharvested central third of the patellar tendon were significantly less than that of controls at both 6 and 12 months. Analysis of collagen fiber size by electron microscopy revealed a significant increase in collagen fiber diameter at 6 months (135 +/- 41 nm versus 49 +/- 4 nm) but no difference between the operative limbs and controls at 12 months. The reharvested bone-patellar tendon-bone complex does not have the same properties as the primary patellar tendon graft up to 1 year after harvest in a canine model, and its use for revision cruciate ligament reconstruction must be carefully reexamined. PMID- 9397266 TI - Anterior cruciate ligament reconstruction with autogenous patellar tendon graft followed by accelerated rehabilitation. A two- to nine-year followup. AB - We sought to determine the long-term results of 1057 consecutive patients who underwent an anterior cruciate ligament reconstruction with an autogenous patellar tendon graft from 1987 through 1993 and who followed an accelerated rehabilitation program. The patients were followed prospectively and objective physical examination data were obtained on 806 patients at a mean of 4.0 years postoperatively. Subjective follow-up data were obtained on 948 patients at a mean of 4.4 years postoperatively. The mean final range of motion was 5 degree/0 degrees/140 degrees. The mean manual maximum KT-1000 arthrometer score was 2.0 +/ 1.5 mm. Isokinetic quadriceps muscle strength testing revealed a mean of 94% strength after acute reconstructions and 91% strength after chronic reconstructions. International Knee Documentation Committee evaluation after acute reconstruction rated 42% of knees as normal, 47% as near normal, 10% as abnormal, and 1% as severely abnormal. The same evaluation after chronic reconstruction rated 41% of knees as normal, 44% as near normal, 14% as abnormal, and 1% as severely abnormal. Radiographically, 94% of acute knees and 89% of chronic knees had no joint space narrowing. Subjective modified Noyes questionnaire results showed a mean score of 93.2 +/- 7.9 points. The mean time for patients to return to sport-specific activities was 6.2 weeks and to athletic competition at full capacity was 6.2 months postoperatively. In the long-term, patients exhibited full range of motion, excellent stability, good strength, and a return of full function in most cases. PMID- 9397267 TI - Experimental study on external tibial rotation of the knee. AB - Using biplanar roentgenographic photogrammetry, we investigated posterolateral rotatory instability of the knee joint both before and after sectioning of posterolateral structures, the posterior cruciate ligament, and the lateral collateral ligament. Fifteen fresh-frozen cadaveric knees were used. Compared with the intact state, sectioning of the posterior cruciate ligament alone did not increase the amount of external tibial rotation, but the axis of external tibial rotation shifted when the anterolateral bundle of the posterior cruciate ligament was cut. When the posterior cruciate ligament was cut after sectioning of the posterolateral structures and the lateral collateral ligament, external tibial rotation increased and the axis of external rotation shifted. The results demonstrated that sectioning of the anterolateral bundle of the posterior cruciate ligament is associated with a change in the location of the axis of tibial rotation. Therefore an isolated posterior cruciate ligament injury can alter the kinematics of the knee joint by changing the axis of external tibial rotation. The present results also demonstrate that the posterior cruciate ligament serves as a kind of secondary restraint to posterolateral rotatory instability in knees with injured posterolateral structures. Helical motion analysis using biplanar roentgenographic photogrammetry is a useful method for evaluating knee kinematics. PMID- 9397268 TI - The combined dynamic and static contributions to subacromial impingement. A biomechanical analysis. AB - Ten human cadaveric shoulders were tested with a dynamic shoulder model simulating physiologic rotator cuff, deltoid, and biceps muscle forces. The combined effect of the muscle forces and acromial structure on subacromial impingement was measured with minimally invasive, miniature pressure transducers. Shoulders with large acromial spurs had significantly greater impingement pressures at the anterolateral acromion in neutral, internal, and external rotation compared with those with flatter acromia. Application of a biceps muscle force reduced anterolateral acromial pressures by 10%. Failure to simulate a supraspinatus force decreased acromial pressure 52% in shoulders with type III acromia in neutral rotation. Without rotator cuff forces applied, the maximum deltoid muscle force required to elevate the arm increased by 17%. Acromial pressures were increased when no rotator cuff forces were applied, but the increases were not significant. After an anterior acromioplasty, pressures decreased by 99% anteriorly. However, failure to achieve a flat surface posteriorly increased pressures in this location, especially with the shoulder in external rotation. Modeling the rotator cuff and deltoid muscle forces demonstrated the importance of the muscular force couple to center the humeral head during elevation of the arm. The inferior forces of the infraspinatus, teres minor, and subscapularis muscles were necessary to neutralize the superior shear force produced by the deltoid and supraspinatus muscles. PMID- 9397269 TI - Two- to five-year followup of arthroscopic Bankart reconstruction using a suture anchor technique. AB - This is a retrospective review of 27 patients (27 shoulders) with recurrent anterior instability who underwent arthroscopic Bankart reconstruction with a suture anchor technique between 1990 and 1993. Average length of followup was 40 months (range, 26 to 64). The average Bankart rating score was 88 (range, 45 to 100) with 70% good-to-excellent results and 30% fair-to-poor results. The average University of California (Los Angeles) shoulder function score was 32 (range, 27 to 35). The average loss of external rotation in abduction was 1 degree. Eight patients (30%) failed the procedure and had recurrent anterior shoulder instability; seven of these had repeat traumatic events. A Pearson chi-square analysis of multiple variables was performed to determine which variables correlated with a successful result. A higher success rate was obtained if the patient had five or fewer dislocations before surgical reconstruction. This technique should be limited to patients not returning to contact sports, or in whom the improved cosmetic results or increased postoperative external rotation of the arthroscopic procedure are valued. PMID- 9397270 TI - Assessment of failed arthroscopic anterior labral repairs. Findings at open surgery. AB - To assess capsulolabral lesions present in patients after unsuccessful arthroscopic procedures, we reviewed the records of 20 patients who had undergone open shoulder procedures after unsuccessful arthroscopic Bankart procedures for chronic shoulder instability. The Bankart lesion had initially been repaired arthroscopically by transglenoid sutures (N = 10), bioabsorbable tacks (N = 7), suture anchors (N = 2), or arthroscopic screws (N = 1). Five of the 20 patients (25%) had reinjuries to the shoulder after the arthroscopic procedure. The average time from the arthroscopic to the open procedure was 17.9 months. Overall, 12 of the 20 patients (60%) had healed Bankart lesions at the time of open surgery. Eight of the 20 patients (40%) were found to have persistent Bankart lesions, and 15 of the 20 patients (75%) were found to have redundant anterior capsules. The presence of a persistent Bankart lesion significantly correlated with postarthroscopic dislocation, and the presence of capsular laxity significantly correlated with postarthroscopic subluxation. We concluded that capsular laxity is difficult to quantify arthroscopically and is present in a significant percentage of patients with chronic traumatic shoulder instability. Failure to successfully treat either the Bankart lesion or capsular laxity at the time of an arthroscopic Bankart procedure may lead to postoperative instability. PMID- 9397271 TI - Posterior tibial tunnel placement to avoid anterior cruciate ligament graft impingement by the intercondylar roof. An in vitro and in vivo study. AB - Recent recommendations to "customize" tibial tunnel placement based on the slope of the intercondylar roof and the amount knee hyperextension were derived from a series of cases with graft impingement by the intercondylar roof. We believe that this impingement is caused by anterior placement of the graft and not by variations of notch anatomy among individual patients. In Phase 1 of this study, we drilled tibial tunnels in the posteromedial aspect of the anterior cruciate ligament "footprint" after the ligament was excised in cadaveric knees. We then passed an impingement rod into the back of the knee joint. Lateral radiographs with the knee in hyperextension were taken of each specimen, and the distance between the superior border of the rod and intercondylar roof was measured. In Phase 2, we prospectively obtained lateral hyperextension radiographs of 75 consecutive knees with anterior cruciate ligament reconstructions and evaluated them for graft impingement based on recently published guidelines. In Phase 1, we found no cases of impingement and an average roof clearance of 8.3 mm. In Phase 2, we noted no cases of severe impingement, 3 cases of moderate impingement (4%), and 72 cases (96%) with no impingement. We conclude that posteromedial tibial tunnel placement alone is adequate to avoid graft impingement in almost all patients. Individualized tibial tunnel placement with specialized tibial guidance systems is not necessary. PMID- 9397272 TI - The strain behavior of the anterior cruciate ligament during squatting and active flexion-extension. A comparison of an open and a closed kinetic chain exercise. AB - The effects of weightbearing (closed kinetic chain) and nonweightbearing (open kinetic chain) exercises on the biomechanical behavior of an injured anterior cruciate ligament or a healing anterior cruciate ligament graft are unknown. To understand the effects of these exercises on the healing graft, we measured the strain behavior of the normal anterior cruciate ligament in human subjects while they performed squatting, a closed kinetic chain exercise, and active flexion extension of the leg, an open kinetic chain exercise. The maximum anterior cruciate ligament strain values obtained during squatting did not differ from those obtained during active flexion-extension. Also, anterior cruciate ligament strain values obtained during squatting were unaffected by the application of elastic resistance intended to increase muscle activity. These findings indicate that squatting, which produces a substantial compressive joint force, does not necessarily protect the anterior cruciate ligament more than active flexion extension of the leg, which is characterized primarily by contraction of the dominant quadriceps muscle. These findings also demonstrate that increasing resistance during the squat exercise does not produce a significant increase in anterior cruciate ligament strain values, unlike increased resistance during active flexion-extension exercise. PMID- 9397273 TI - Osteochondritis dissecans of the femoral condyles. Long-term results of excision of the fragment. AB - Nineteen patients with 20 osteochondritis dissecans lesions were evaluated between 2 and 20 years after excision of a partially detached (grade III) or loose (grade IV) fragment from the femoral condyles. Evaluation with the Hughston rating scale for osteochondritis dissecans revealed one excellent result, four good, four fair, six poor, and five failure results. Eleven patients had developed osteochondritis dissecans before skeletal maturity. In contrast to what has been stated in the literature, the results in these patients were no better than in those who developed osteochondritis dissecans as adults. The short-term results of excision are good, but the long-term results are extremely poor. Consequently, we recommend bone grafting and replacement of the fragment when it is technically possible because the long-term results are better than those after excision. PMID- 9397274 TI - Results of percutaneous longitudinal tenotomy for Achilles tendinopathy in middle and long-distance runners. AB - From August 1989 to January 1995 we performed multiple percutaneous longitudinal tenotomies under local anesthetic on 52 middle- and long-distance runners with unilateral Achilles tendinitis or peritendinitis that had failed conservative treatment. Forty-eight patients were reviewed at an average of 22.1 months (SD, 6.5) after surgery. Results were rated as excellent in 25 patients, good in 12, fair in 7, and poor in 4. Four patients developed subcutaneous hematomas. One patient developed a superficial infection at one of the incision sites, which was managed by oral antibiotics with full recovery. Three patients complained of over sensitivity to the incisions; this was resolved by rubbing hand cream over the incisions several times a day. One patient developed hypertrophic painful scars on three of the five incisions, but corticosteroid injections yielded good functional and cosmetic results. Isometric strength and endurance of the gastrocsoleus complex was measured just before the procedure, and at 6 weeks and 6 months later. Both were within 10% of the normal contralateral limb by the 6th postoperative month. Percutaneous longitudinal tenotomy is simple, can be performed on an outpatient basis, requires minimal follow-up care, and, in our experience, has produced no significant complications. We use this procedure as the operative treatment of choice for cases of chronic tendinitis that have failed conservative treatment. PMID- 9397275 TI - Tissue shrinkage with the holmium:yttrium aluminum garnet laser. A postoperative assessment of tissue length, stiffness, and structure. AB - The effect of laser energy on the length, stiffness, and structure of connective tissue was examined in a rabbit patellar tendon model. A holmium:yttrium-aluminum garnet laser was used to deliver a calculated dose of laser energy (300 J/cm2) to one randomly selected patellar tendon in each of 13 adult New Zealand White rabbits. The contralateral patellar tendon was used as a control. Radiopaque markers were placed in the patella and tibial tuberosity to allow for patellar tendon length measurements (via standard lateral radiographs) before and after laser application and at 4 and 8 weeks. Limbs were not immobilized during the postoperative period. The tendons were harvested at 0 weeks (N = 7) and 8 weeks (N = 6) and evaluated for tensile, stiffness, cross-sectional area, histologic changes, and electron microscopic appearance. The results demonstrated significant tendon shrinkage (6.6% +/- 1.4%) after application of the calculated laser energy dose. However, tendon length had increased significantly beyond the immediate postlaser length at 4 weeks and beyond its original length by 8 weeks. At 8 weeks, the lased tendons were significantly less stiff with significantly greater cross-sectional areas than contralateral controls. There was generalized fibroblastic response throughout the entire lased tendon characterized by a marked increase in cellularity. There was also a change from the normal bimodal pattern of large- and small-diameter collagen fibers to a unimodal pattern with predominantly small-diameter fibers in the lased tendons. The tissue alterations seen in this study suggest that the biologic response of connective tissue to laser energy causes a further compromise in tissue integrity, beyond that attributed to the initial physical effects of the laser. These alterations must be taken into consideration when determining postoperative rehabilitation of laser-modified tissues. PMID- 9397276 TI - Popliteomeniscal fasciculi and lateral meniscal stability. AB - In an attempt to understand better the contribution of the anteroinferior and posterosuperior popliteomeniscal fasciculi to lateral meniscus stability, we objectively evaluated the stability of the lateral meniscus before and after sequentially sectioning these fasciculi. In the biomechanical model, we attempted to account for the inherent limitations of arthroscopic evaluation of lateral meniscal stability. When the fasciculi were intact, the average lateral meniscal motion with a 10-N load was 3.6 mm. When the anteroinferior fascicle was disrupted, the average lateral meniscal motion with a 10-N load was 5.4 mm. The mean increase in motion from the intact state was 1.8 mm or 50%, which was significant. When both fasciculi were disrupted, the average lateral meniscal motion with 10-N load was 6.4 mm. The mean increase in motion from the intact state was 2.8 mm or 78% and from the single fascicle disruption state was 1.0 mm or 18%, both differences were significant. The meniscus did not become locked with any of these loading trials, and it spontaneously reduced to the original position when unloaded. Both fasciculi make significant contributions to meniscal stability. Even though the meniscus never became locked in the joint when loaded during this study, with the variable loads seen with normal activities mechanical symptoms might be expected when meniscal motion is almost double. An increase in lateral meniscal motion at the time of surgery may aid in the diagnosis of fasciculi disruption, despite normal meniscal structure on magnetic resonance images and at arthroscopic visualization. PMID- 9397277 TI - Adventitial cystic disease of the popliteal artery in a triathlete. A case report. PMID- 9397278 TI - Functional evaluation of the ligaments at the acromioclavicular joint during anteroposterior and superoinferior translation. AB - We examined the anatomy and measured the in situ force in ligaments at the acromioclavicular joint using a universal force-moment sensor. The in situ force in the coracoacromial, conoid, trapezoid, superior acromioclavicular capsular, and inferior acromioclavicular capsular ligaments of 10 fresh-frozen cadaveric shoulders was determined for a load of 70 N applied to the clavicle in anteroposterior and superoinferior directions. The lengths of the conoid and trapezoid ligaments were found to be 15.1 +/- 4.1 and 11.5 +/- 2.2 mm, respectively; the widths of the conoid and trapezoid ligaments were 10.7 +/- 1.5 and 11.0 +/- 2.8 mm, respectively. The in situ force of the trapezoid (42.9 +/- 15.4 N) was significantly greater than that for the other ligaments during posterior displacement. Otherwise, no statistically significant differences could be found between any of the in situ forces in each ligament during all other motions examined. During anterior displacement, the inferior acromioclavicular capsular ligament appeared to be the major restraint. The trapezoid ligament was the primary restraint during posterior displacement and provided 55.8% +/- 20.0% of the resisting force. Our results suggest that the coracoclavicular and other acromioclavicular joint capsular ligaments should be considered for reconstruction to restore normal joint function, especially in the anterior, posterior, and superior directions. PMID- 9397279 TI - The clinical importance of the anaerobic energy system and its assessment in human performance. AB - The anaerobic energy system is involved in providing energy for all forms of physical activity. The relevance of this system to human performance and physical fitness throughout the age spectrum is underscored here and contrasted with the aerobic energy system. The anaerobic system responds to high-intensity training with biochemical, neural, and anatomic adaptations. Unlike the aerobic system, this response tends to be primarily a local phenomenon with little systemic adaptation. An important factor distinguishing anaerobic training from aerobic training is the intensity of the exercise dose. For anaerobic training to occur, the dose must be of high intensity and performed to near-exhaustion. The anaerobic system can be indirectly assessed by performance tests, such as a vertical jump or stair climb, or more directly by supramaximal bicycle tests. The impact of recent research regarding the trainability of the anaerobic system, particularly in the elderly population, is encouraging. The elderly respond to anaerobic training and, as a result, their independence, quality of life, and safety from falls can be improved. While little is known about anaerobic rehabilitation after injury, it is known that isokinetic and performance tests may be considered normal after rehabilitation, despite incomplete rehabilitation of the anaerobic system. Thus, appropriate application of the anaerobic system assessments and training principles is an important aspect of sports medicine practice. PMID- 9397280 TI - Athletics and osteoarthritis. AB - Athletes, and an increasing number of middle aged and older people who want to participate in athletics, may question whether regular vigorous physical activity increases their risk of developing osteoarthritis. To answer this, the clinical syndrome of osteoarthritis must be distinguished from periarticular soft tissue pain associated with activity and from the development of osteophytes. Sports that subject joints to repetitive high levels of impact and torsional loading increase the risk of articular cartilage degeneration and the resulting clinical syndrome of osteoarthritis. However, moderate habitual exercise does not increase the risk of osteoarthritis; selected sports improve strength and mobility in older people and people with mild and moderate osteoarthritis. People with abnormal joint anatomy or alignment, previous significant joint injury or surgery, joint instability, above-average body weight, disturbances of joint or muscle innervation or inadequate muscle strength probably have increased risk of osteoarthritis. These people and those with early osteoarthritis can benefit from regular physical activity, but they should have a careful evaluation of their joint structure and function before participation. They should consider measures that decrease the intensity and frequency of impact and torsional loading of joints, including use of sports equipment that decreases joint impact loading, maintaining or improving muscle strength, tone, and general conditioning so that muscle contractions help protect joints from injury and high impact, and decreasing body weight. PMID- 9397281 TI - A statistics primer. Tests for continuous data. PMID- 9397283 TI - Perceived threat of illness and health protective behaviors in offspring of adults with non-insulin-dependent diabetes mellitus. AB - Self-reported measures of perceived threat of illness, health protective behaviors, psychological well-being, and family modeling of health behaviors of 30 adults with a parental history of non-insulin-dependent diabetes mellitus (NIDDM) were compared with responses from 29 adults with a parental history of hypertension and 30 adults with no parental history of chronic illness. The NIDDM risk group reported significantly more perceived threats of NIDDM and hypertension and more weight-control efforts than the controls did. Reports of the NIDDM risk respondents concerning physician screening, healthy diet, and exercise did not differ from reports of individuals without a family history of NIDDM. Perceived threat, psychological well-being, and family modeling did not correlate with health-protective behaviors. The findings suggest that offspring of adults diagnosed with NIDDM perceive themselves to be at risk of NIDDM and engage in health behaviors, such as weight control, to protect themselves from NIDDM onset. PMID- 9397282 TI - The relationship between alcohol, stress, and depression in Mexican Americans and non-Hispanic whites. AB - The effect of alcohol use on the relationship between stress and depression in US born Mexican American men, Mexican Americans born in Mexico, and non-Hispanic Whites born in the United States was examined in a sample obtained from the Los Angeles Epidemiological Catchment Area study. Chronic stress, measured by financial strain, and acute stress, measured by negative life events, were included in the analysis. Alcohol use was measured through a combination of frequency, quantity, and binging behavior. Non-Hispanic Whites were found to have a U-shaped relationship in which moderate drinkers, in the presence of stress, had lower levels of depression than did heavy drinkers and abstainers. No such U shaped relationship for Mexican Americans born in the United States was indicated. Mexican Americans born in Mexico had a more J-shaped relationship, with abstainers through moderate drinkers having lower mean depression scores than did heavy drinkers. PMID- 9397284 TI - A field study of stress and fatigue in long-distance bus drivers. AB - Psychophysiological changes during long-distance driving may be associated with driving fatigue and morbidity. Measures of stress and arousal, including heart rate, blood pressure, catecholamines, cortisol, state anxiety, and self-ratings of stress and arousal were collected from 10 long-distance bus drivers during 12 hour driving shifts and at matched times on nondriving rest days. Cardiovascular and catecholamine data were elevated across the entire work day, compared with rest days. Self-reported stress and state anxiety were elevated only at the preshift measure, and these elevations were interpreted as the result of anticipatory anxiety and additional work demands at the beginning of the shift. Decelerating activation from the 9th to the 12th hours of driving were reflected in slower heart rate and lower subjective arousal ratings. Suggested explanations for these findings are that drivers experience a release of tension when they anticipate the end of the shift and therefore deactivation is a signal or precursor to the onset of fatigue in physiological adjustment mechanisms. PMID- 9397285 TI - Maintaining attendance at a fitness center: an application of the decision balance sheet. AB - The effect of a decision balance sheet intervention on attendance at a university fitness facility was examined. Facility members were randomly assigned to control, placebo, and experimental conditions. The control condition received no intervention, whereas the placebo and experimental conditions were called by telephone and asked to complete either an irrelevant (smoking) or relevant (exercising at the fitness facility) decision balance sheet. Attendance was monitored surreptitiously for 4 weeks baseline and 8 weeks post intervention. Statistical analyses indicated that the control and placebo conditions showed a significant decrease in attendance from baseline to intervention, whereas those in the experimental condition maintained attendance levels. Discussion focused on broadening the application of the decision balance sheet, determining its theoretical boundaries, and the necessity and appropriateness of decision alternatives for the decision balance sheet in the exercise domain. PMID- 9397286 TI - Self-efficacy and adjustment in cancer patients: a preliminary report. AB - The relation between cancer self-efficacy and patient cancer adjustment, depression, psychological distress, and behavioral dysfunction in 42 cancer patients was studied in a preliminary investigation. Participants were male cancer outpatients recruited from a Veterans Administration Medical Center who completed a Cancer Self-Efficacy Scale, the Center for Epidemiological Studies Depression Scale, the Affect Balance Scale, and the Sickness Impact Profile. Correlational analyses indicated that self-efficacy was related to all adjustment measures. Regression analyses revealed that when age, education, time since initial diagnosis, and current disease status were controlled, the relationships between patient self-efficacy expectations and cancer adjustment, psychological distress, negative affect, positive affect, and behavioral dysfunction remained statistically significant. Taken together, the results of the analyses suggested that patient expectancies about control over cancer-related symptoms were related to several important aspects of patient functioning. The results underscored the need for further investigation of this construct in cancer patients. PMID- 9397287 TI - Safety of calcium channel blockers: perspective on the controversy. PMID- 9397288 TI - The endothelium in coronary artery disease. AB - An increasing body of evidence indicates that the endothelium is crucially involved in the regulation of coronary blood flow and cardiac function. Injury to the endothelium precipitates atherosclerosis by leading to smooth-muscle-cell migration and proliferation, induction of expression of growth factors and impairment in the plasmatic coagulation and endogenous fibrinolysis system. Strategically located between the circulating blood and the vascular smooth muscle, endothelial cells release numerous vasoactive substances regulating the function of vascular smooth muscle and trafficking blood cells. Important endothelium-derived vasodilators are prostacyclin, bradykinin, nitric oxide and, independent of the former, endothelium-derived hyperpolarizing factor. In particular, nitric oxide inhibits cellular growth and migration. In concert with prostacyclin, nitric oxide exerts potent antiatherogenic and thromboresistant properties by preventing platelet aggregation and cell adhesion. These effects are counterbalanced by endothelial vasoconstrictors, such as angiotensin II and endothelin-1, both of which exert prothrombotic and growth-promoting properties. Modern therapeutic strategies in coronary artery disease focus on preserving or restoring endothelial integrity. Whereas nitrates partly substitute deficient endogenous nitric oxide, calcium antagonists counteract angiotensin II and endothelin-1 at the level of vascular smooth muscle by reducing Ca2+ inflow and facilitating the vasodilator effects of nitric oxide. Beyond inhibiting the renin angiotensin system, angiotensin-converting enzyme inhibitors diminish the inactivation of bradykinin, thus leading to an augmentation of nitric oxide release. Furthermore, newly developed specific endothelin antagonists will provide us with greater insight into the beneficial effects of restoring endothelial dysfunction in cardiovascular disease. Thus, drugs can directly affect endothelial function, prevent the action of endothelial mediators, substitute for deficient endothelial factors or indirectly exert protective effects by interfering with cardiovascular risk factors. PMID- 9397289 TI - Structure and function of small arteries of essential hypertensive patients following chronic treatment with once-a-day nifedipine. AB - In view of the important impact of small-artery structural and functional abnormalities on complications of hypertension and recent data suggesting that some antihypertensive agents may correct some of these abnormalities, a study of resistance artery structure and function in 20 well-controlled essential hypertensive patients who had received for a prolonged period of time monotherapy with the once-a-day extended release formulation of the calcium channel antagonist nifedipine (nidefipine GITS) or with the beta-blocker atenolol is reviewed. Resistance-size small arteries (standardized lumen diameter of 247 +/- 8 microns) were studied after dissection from a gluteal subcutaneous biopsy. Small arteries were investigated on a wire myograph and as pressurized vessels. On the myograph, the media width-to-lumen diameter ratio of arteries was 5.37 +/- 0.09% in normotensive subjects, 5.38 +/- 0.18% in patients treated with nifedipine GITS, 6.81 +/- 0.18% in patients treated with atenolol and 7.08 +/- 0.12% in untreated hypertensives (p < 0.001, untreated or atenolol-treated patients vs. normotensives or nifedipine-GITS-treated hypertensives), and similar results were found in pressurized arteries. Contractility and endothelium dependent relaxation were impaired in small arteries from untreated or atenolol treated patients in comparison to those from normotensive subjects or nifedipine GITS-treated patients. In conclusion, hypertensive patients with well-controlled blood pressure under treatment for more than 1 year with nifedipine GITS exhibit normal structure and function of small arteries, whereas similar patients whose blood pressure is as well controlled by the beta-blocker atenolol present abnormally thick small arteries with impaired contractility and endothelium dependent relaxation. It will be important to determine whether small arteries of other vascular beds are also improved by nifedipine GITS treatment of elevated blood pressure and whether this results in reduced morbidity and mortality in hypertensive patients. PMID- 9397291 TI - Primary prevention of cardiovascular disease. AB - Antihypertensive drug treatment has been shown to significantly decrease cardiovascular morbidity and mortality rates in hypertensive subjects and to reduce the occurrence of major hypertension-related complications, such as stroke, coronary artery disease, congestive heart failure and renal insufficiency. Despite these favorable effects, several issues related to antihypertensive treatment remain to be clarified. This paper will discuss some of these issues, with particular reference to the effects of blood pressure lowering on renal diseases and coronary heart disease. It will also discuss the ongoing clinical trials aimed at clarifying some unsolved issues of antihypertensive treatment. The objective of the International Nifedipine GITS Study Intervention as a Goal in Hypertensive Treatment was to provide information on the effects of calcium antagonist treatment on high blood pressure values and on hypertension-related cardiovascular complications in a high-risk hypertensive population. PMID- 9397290 TI - Optimal treatment of stable angina. AB - Angina pectoris due to coronary artery disease is a common manifestation of myocardial ischemia. Reduction of oxygen demand (beta-blockers) and relief of coronary vasoconstriction (calcium blocker or nitrate) are additive approaches to controlling ischemia. Risk factor reduction may improve coronary vascular physiology, and ASA reduces the likelihood of thrombosis and myocardial infarction. It is still unclear whether reduction of angina reduces cardiac morbidity and/or mortality. In the Asymptomatic Cardiac Ischemia Pilot Study (ACIP) and Total Ischemic Burden Bisoprolol Study (TIBBS) trials, data suggest benefit from reducing myocardial ischemia. Thus control of angina pectoris is a major goal of the treatment of coronary artery disease. PMID- 9397292 TI - Equivalent reduction of proteinuria in hypertensives by either nifedipine GITS or enalapril: disparate effects on neurohormones and ambulatory blood pressure and the influence of salt. AB - OBJECTIVE: We compared the efficacy of two classes of antihypertensive therapy on ambulatory blood pressure control and proteinuria in patients with hypertension. Furthermore, we determined the effects of the interaction of these therapies on neurohormonal activation and of the patients' ambient sodium intake on the outcomes. METHODS: Sustained-release nifedipine (nifedipine gastrointestinal therapeutic system, GITS) 30-120 mg/day was compared in a double-blind sequential randomized placebo-controlled trial with enalapril 5-30 mg/day regarding office and 24-hour blood pressure control, plasma renin activity, noradrenaline and adrenaline levels and 24-hour urinary protein and sodium in 46 elderly nondiabetic hypertensive patients in a 16- to 18-week trial. RESULTS: Both nifedipine GITS and enalapril controlled ambulatory blood pressure during the day and at peak effect. Nifedipine GITS controlled ambulatory blood pressure during the early morning surge and at night time as well. Nifedipine GITS increased plasma renin activity and noradrenaline by 50 and 20%, respectively, compared to the 150 and 0% change produced by enalapril. Both nifedipine GITS and enalapril reduced proteinuria by 37%. Patients had increasing levels or proteinuria proportional to higher ambient sodium intake (r = 0.48; p < 0.01). This effect was accentuated during nifedipine GITS therapy as compared to enalapril. CONCLUSION: Nifedipine GITS was superior to enalapril in controlling ambulatory blood pressure, but they were equivalent in reducing proteinuria (37%). They had disparate effects on neural activation and the duration of action. Raised protein excretion appears to be associated with raised sodium intake. This was apparent especially during nifedipine XL therapy. PMID- 9397293 TI - Nifedipine GITS replacing nifedipine SR: ambulatory blood pressure assessment of efficacy. AB - The relative efficacy of two formulations of nifedipine, slow release (SR) and gastrointestinal therapeutic system (GITS), to lower blood pressure in hypertensive patients was evaluated in a prospective study. Nifedipine GITS 30 mg/day replaced nifedipine SR, 20 mg b.i.d. in 38 patients, 23 monitored by routine blood pressure measurements and 15 by ambulatory monitoring. Nifedipine GITS achieved a marked reduction in blood pressure with a smaller dose than nifedipine SR: 30 versus 40 mg/day, respectively. It is concluded that patients with hypertension controlled on a twice-daily dose of nifedipine prolonged action can be converted to a lower once-daily dose of nifedipine GITS without experiencing any increase in blood pressure. Tolerability and compliance improved when switching to the GITS formulation. PMID- 9397294 TI - Interaction of antihypertensive drugs with anti-inflammatory drugs. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) can induce an increase in blood pressure (BP) and may potentially reduce the efficacy of several antihypertensive drugs. Probably the main mechanism of action is inhibition of prostaglandin (PG) synthesis since NSAIDs have higher propensity to increase BP as the regulation of BP (and renal function) is more PG-dependent and to interact with drugs (diuretics, beta-blockers and ACE inhibitors) that may act through the increase of PG formation. In contrast, NSAIDs do not interact with calcium antagonists and central acting drugs which actions are apparently unrelated with renal/extrarenal production of PG. It has been claimed that inhibition of natriuretic PGs could explain the pressure effects of NSAIDs in treated hypertensive patients, but sodium retention may be not the single explanation for such an interaction. We found that despite indomethacin produced sodium retention after being added either to enalapril or nifedipine-GITS, it only attenuated (by 45%) the antihypertensive effects of enalapril. In alternative, since PG enhances vasodilatation and attenuates vasoconstrictor influences, some NSAIDs may counteract the PG-dependent vasodilatory tone in renal and extrarenal vascular beds that mediate the antihypertensive action of some drugs. Thus, since calcium antagonists are probably not affected by NSAIDs, they may be preferable to drugs like diuretics, beta-blockers and ACE inhibitors for the treatment of high blood pressure control in hypertensive patients who are clinically suitable for NSAIDs therapy. PMID- 9397295 TI - Anti-atherosclerotic properties of nifedipine. Benefit of early intervention to prevent cardiovascular complications. AB - Early stages of the atherosclerotic plaque are considered to be responsible for about 2/3 of all acute coronary syndromes. Therefore, suppression of this initial stage of plaque formation constitutes a major target to reduce the incidence of the disease itself as well as its complications. Ca antagonists, like Nifedipine, interfere with Ca++ ions crucially involved in atherogenesis. Consequently, interval angiographic trials with Nifedipine in patients afflicted with coronary artery disease assessed a significant reduction of coronary lesions. Clinical outcome trials will establish the prognostic importance of the anti atherosclerotic properties of Nifedipine. PMID- 9397296 TI - Renal protection in diabetes--an emerging role for calcium antagonists. AB - The combination of diabetes and hypertension increases the changes of progressive renal disorder and ultimately renal failure. Roughly 40% of all diabetics, whether insulin dependent or not, develop diabetic nephropathy. Diabetic nephropathy is the single most important cause of end-stage renal disease in the western world and accounts for more than a quarter of all end-stage renal diseases. It is also a major cause of increased morbidity and mortality in diabetic patients. Increased arterial blood pressure is an early and common phenomenon in incipient and overt diabetic nephropathy. The relationship between arterial blood pressure and diabetic nephropathy is a complex one, diabetic nephropathy increasing blood pressure and blood pressure accelerating the course of nephropathy. Calcium antagonists antagonize preglomerular vasoconstriction. Furthermore, additional putative mechanisms include the ability to retard renal growth and possibly to attenuate mesangial entrapment of macromolecules and to attenuate the mitogenic effects of diverse growth factors. Calcium antagonists (except the old short-acting dihydropyridine drugs) reduce microalbuminuria and preserve kidney function in diabetic patients with incipient diabetic nephropathy. Long-term trials using the new long-acting dihydropyridine calcium antagonists for the treatment of patients with incipient nephropathy are still lacking. A recent 1-year randomized double-blind study in hypertensive insulin dependent diabetic patients with diabetic nephropathy showed a more beneficial effect on the decline rate in the glomerular filtration rate of nisoldipine (long acting dihydropyridine) than angiotensin-converting-enzyme (ACE) inhibition. The mean arterial blood pressure during the study based on 24-hour recordings was nearly identical, 103 (SD 9) and 101 (SD 11) mm Hg in the two groups. Furthermore, a recent 5-year randomized open study in hypertensive noninsulin dependent patients with diabetic nephropathy has revealed the same beneficial effect of a calcium antagonist and ACE inhibition on the progression of nephropathy. In a third group treated with sympatholytic drugs, more than 50% of the subjects had a doubling of their creatinine as compared to less than 10% in the two other groups mentioned above. However, long-term studies are needed to consolidate these findings and expand them to insulin-dependent diabetic patients with diabetic kidney disease. PMID- 9397297 TI - Evaluating the association between drug use and outcome--should the information from observational studies affect therapeutic use? PMID- 9397298 TI - Calcium channel blockers and cancer: is there preclinical evidence for an association? AB - The preclinical evidence for a potential influence of calcium channel blockers (CCB) on carcinogenesis is discussed in the light of rodent carcinogenicity studies as well as mechanistic data. In the bioassays performed in rats and mice on the dihydropyridine CCB nifedipine, nimodipine, nisoldipine and nitrendipine, no evidence was found for a carcinogenic potential of these compounds. Moreover, the mechanistic knowledge on the influence of CCB on the fundamental processes of cell proliferation and apoptosis is not in favor of a tumor-promoting activity of these compounds. It is, therefore, concluded that there is no preclinical evidence that the therapeutic use of CCB of the dihydropyridine class is associated with an increased risk of cancer. PMID- 9397299 TI - Elevated serum cardiac markers in asymptomatic marathon runners after competition: is the myocardium stunned? AB - Prolonged strenuous exercise may trigger acute myocardial infarction (AMI), as exemplified by the occurrence of sudden cardiac death during marathon running. Serum creatine kinase MB (CK-MB) may be elevated in asymptomatic marathon runners after competition from exertional rhabdomyolysis of skeletal muscle altered by training, limiting its utility for evaluating acute cardiac injury in such athletes. Myoglobin and CK-MB2 isoform levels are emerging as earlier markers of AMI and troponin subunits as more specific than serum CK-MB mass. We tested runners before and sequentially after the 1995 Boston Marathon for conventional and newer markers including myoglobin, CK-MB mass and isoforms, cardiac troponin T, and cardiac troponin I using standard laboratory methods and rapid format assays if available. The mean serum values for myoglobin, CK-MB mass, CK MB/myoglobin rapid panel tests, and CK-MB2 isoforms were normal or negative pre race and elevated or positive 4 and 24 h after competition. These markers lack specificity for acute cardiac injury in trained runners. While the mean serum values for cardiac troponins T and I remained normal, 9 of 45 runners (20%) showed an increase in subunits by first-generation assays. All runners remained asymptomatic for cardiac disease and completed subsequent marathons 1 year later, making reversible myocardial injury or stunning unlikely. Elevated values of serum markers for AMI, including first-generation assays for both troponin subunits should be interpreted with caution in trained runners. PMID- 9397300 TI - Short- and long-term prognosis after coronary artery bypass grafting in relation to smoking habits. AB - We describe the 2- and 5-year prognoses following coronary artery bypass grafting (CABG) in relation to smoking habits among consecutive patients being operated on in western Sweden during a 3-year period. Among the 2,121 patients, 10.2% admitted smoking at coronary angiography as compared with 7.5% 2 years after CABG (NS). Among smokers, the mortality during the subsequent 2 years was 8.9% as compared with 6.5% for exsmokers and 7.3% for never smokers (NS). During the 5 year follow-up, smokers had a mortality of 18.8% as compared with 13.6% for exsmokers and 12.5% for never smokers (p = 0.03). When correcting for dissimilarities in previous history, smoking was a strongly significant independent (p < 0.0001) predictor of 5-year mortality. PMID- 9397301 TI - Doppler echocardiographic assessment of left ventricular diastolic function in myotonic dystrophy. AB - We utilized Doppler echocardiography to characterize left ventricular diastolic function in 42 patients with myotonic dystrophy (mean age 37 +/- 12 years, 64% male) who had no symptoms of heart failure and had normal left ventricular systolic function. Data were compared with those in 41 normal control subjects of similar age and gender. Heart rate, systemic blood pressure, and cardiac dimensions (wall thickness, left atrial and left ventricular cavity dimensions) were similar and not significantly different in patients and controls. As a group, patients showed significantly increased deceleration time and decreased rate of decline of flow velocity in early diastole (p < 0.0001 and p < 0.01, respectively) when compared to controls. Individual patient analysis showed that 10 (24%) of the 42 patients with myotonic dystrophy had 2 or more abnormal Doppler indexes of diastolic function consistent with a pattern of impaired left ventricular relaxation. The most common abnormalities were increased deceleration time (> 224 ms; 9 patients), prolonged isovolumic relaxation time (> 103 ms; 8 patients) and reduced rate of decline of flow velocity in early diastole (< 2.1 m/s2; 5 patients). In addition, peak early diastolic flow velocity was reduced (< 43 cm/s) in 3 patients and early to atrial peak flow velocity ratio was reduced (< 1) in 2 patients. Comparison of subgroups of patients with and without abnormal Doppler indexes showed no significant differences with regard to age, gender, heart rate, systemic blood pressure, severity of neuromuscular disease, and cardiac dimensions. After study, patients were clinically followed up for a mean period of 20 +/- 7 months (range 12-35). During observation no patients died and none experienced symptoms of heart failure. This Doppler echocardiographic analysis demonstrates that diastolic abnormalities may be present in patients with myotonic dystrophy, even in the absence of symptoms of cardiac failure or left ventricular systolic dysfunction. These diastolic abnormalities suggest an intrinsic myocardial abnormality in patients with myotonic dystrophy; however, whether they represent a preclinical phase of myocardial involvement or an intrinsic feature of the primary myocardial disease process in myotonic dystrophy remains to be elucidated. PMID- 9397302 TI - Hyperinsulinemia in patients with spastic angina pectoris. AB - We evaluated the association between coronary spasm and hyperinsulinemia (high immunoreactive insulin, IRI) in patients with angina pectoris. The study cohort comprised 30 patients with spastic angina pectoris, 30 patients with angina pectoris showing fixed-obstructive coronary sclerosis and 30 control subjects who were matched for body mass index, age and sex. A 75-gram oral glucose test was performed, and blood sugar and IRI were serially measured concomitant with serum total cholesterol, triglyceride and HDL cholesterol. The IRI level at 60 min, the peak IRI during the test, sigma IRI and sigma IRI/sigma blood sugar were significantly higher in the patients than in the controls. Total cholesterol and LDL cholesterol levels were significantly increased in patients showing fixed obstructive coronary sclerosis compared to controls. PMID- 9397303 TI - Daily distribution of episodes of acute cardiogenic pulmonary edema. AB - Many fatal or potentially fatal cardio-cerebrovascular diseases present a definite circadian distribution in their onset. In order to verify if episodes of acute cardiogenic pulmonary edema have a significant daily periodicity in their occurrence, a retrospective analysis of 1,204 episodes has been conducted. In all cases, the hour of the day of onset has been identified with certainty; all episodes occurred in hospitalized patients. The rhythmometric circadian inferential statistical analysis by means of the single cosinor method demonstrates that the episodes of acute cardiogenic pulmonary edema present a significant (p < 0.002) circadian distribution, with a peak at 1:00 a.m. (from 10:00 p.m. to 4:00 a.m.). No significant differences (p > 0.05) were found in the circadian distribution regarding sex, age (less or more than 60 years), absence or presence of arterial hypertension and coronary artery disease. Several factors may contribute to this behavior, especially the relationships between several endogenous circadian rhythms, sleep and disease. The knowledge that acute pulmonary edema is a high chronorisk disease could be of interest for the better understanding of its pathophysiology and for a better causative control and prevention. PMID- 9397304 TI - Cardiac involvement in Behcet's disease. AB - To assess the prevalence and the extent of cardiac involvement in patients with Behcet's disease and to investigate the possible causes that may predispose to this involvement, 30 patients affected by Behcet's disease and 30 normal control subjects were submitted to M-mode, two-dimensional, and Doppler echocardiographic evaluation. Moreover, antinuclear and anticardiolipin autoantibodies were determined in the sera of both patients and control subjects. Finally, HLA-B51 positivity was assessed in the patients and in a historical control group. Mitral valve prolapse was observed in 50% and proximal aorta dilatation in 30% of the patients. There was a significant difference in the rate of these abnormalities in comparison with the control group. Left ventricular function parameters were similar between the two groups. The positivity rate of antinuclear and anticardiolipin autoantibodies was very low (7%), without differences between the groups. HLA-B51 was detected in 82.7% of the patients versus 21.7% in the control group (p < 0.00001). In conclusion, this study demonstrates a high rate of cardiac abnormalities in patients with Behcet's disease. PMID- 9397305 TI - Protein turnover in compensated chronic aortic regurgitation. AB - We recently demonstrated dynamic alterations in protein turnover 3 days and 1 month after surgical induction of aortic regurgitation (AR). To characterize protein synthesis and degradation during the long-term plateau phase, we performed [3H]-leucine infusion 2.5 years after induction of AR in 10 New Zealand White rabbits and 12 sham-operated controls. Protein fractional synthesis rates were obtained by analyses of plasma and protein hydrolysates, growth rates from protein concentration and heart weight measurements, and degradation rates by subtraction of growth from synthesis rates. AR (regurgitant fraction 25 +/- 11%) caused a 57% increase in left ventricular (LV) weight in comparison with controls (7.4 +/- 1.7 vs. 4.7 +/- 0.6 g, p < 0.001) and no evidence of heart failure. Although concentrations of total cardiac protein, myosin heavy chain and actin were similar, the enlarged AR hearts had increased amounts of total cardiac protein (1,009 +/- 312 vs. 682 +/- 120 mg/LV, p < 0.05), myosin heavy chain (148 +/- 91 vs. 81 +/- 29 mg/LV, p < 0.05), and actin (73 +/- 42 vs. 44 +/- 16 mg/LV, p < 0.06). Individual protein fractional synthesis and degradation rates were closely balanced. However, myosin fractional synthesis rates were 152% (p < 0.01) greater than those of total cardiac protein in AR animals, while only 52% (p < 0.05) greater in controls (AR vs. controls, p = 0.05). Variations in actin turnover between AR and control animals did not attain statistical significance. Myosin and actin fractional synthesis rates correlated closely in AR rabbits (R = 0.81, p < 0.02), but not among controls (R = 0.41, NS). Thus, selective alterations in myofibrillar protein turnover contribute to the maintenance of increased myofibrillar protein content in the 'compensatory' LV hypertrophy of chronic AR. PMID- 9397306 TI - Effects of antegrade versus combined antegrade/retrograde cardioplegia on postoperative septal wall motion in patients undergoing open heart surgery. AB - To evaluate the influence of two techniques of myocardial protection on septal wall motion (SWM) and left ventricular ejection fraction, 21 patients with a normal SWM underwent surgery using either conventional antegrade cardioplegia (group I, n = 9) or combined antegrade/retrograde cardioplegia (group II, n = 12). The patients were assessed pre- and postoperatively by radionuclide ventriculography. A resting thallium-201 study was performed in patients (n = 6) with a postoperatively abnormal SWM: in 2 of 9 (22%) in group I and in 4 of 12 (33%) in group II (p = NS). The left ventricular ejection fraction was similar in both groups before surgery (57 +/- 3% in group I vs. 57 +/- 8% in group II; p = NS) and did not change significantly after surgery. All 6 patients with an abnormal SWM had a normal septal thallium-201 uptake. Thus, (1) an abnormal SWM after cardiac surgery is common: (2) it is not due to perioperative ischemia or infarction, and (3) neither the incidence of an abnormal SWM not the global left ventricular function is influenced by the addition of retrograde cardioplegia during open heart surgery. PMID- 9397307 TI - Dislodgement of a Wiktor stent during intracoronary ultrasound examination. AB - We report a case of stent dislodgement complicating adjuvant intracoronary ultrasound (ICUS) imaging that required emergency coronary bypass grafting. This probably very rare complication gains importance since ICUS is increasingly used to confirm adequate stent expansion and full coverage of the lesion. PMID- 9397308 TI - Early detection of myocardial ischemia after successful percutaneous coronary angioplasty. AB - We evaluated the functional significance of angiographically successful percutaneous transluminal coronary angioplasty (PTCA) in 50 patients before and after PTCA using an atrial pacing stress test. Before balloon angioplasty, 40/50 patients had transient ST-segment changes on the intracoronary (IC) ECG. After PTCA 14/50 patients continued to have ischemic changes on IC-ECG. Atrial pacing stress tests can be performed easily in the cardiac catheterization laboratory. Despite angiographically successful dilatation, 28% of the patients have inducible ischemia indicating functionally inadequate dilatation. Inadequate functional dilatation may contribute to early return of symptoms in some patients. PMID- 9397309 TI - Clinical significance of the urinary oxygen tension in patients with ischemic heart disease. AB - The clinical significance of the urinary oxygen tension (PuO2) was evaluated in 60 patients with ischemic heart disease. The PuO2 had fair relations to cardiac index and serum creatinine level (r = 0.73 and r = 0.73, respectively). Although the PuO2 had a fair relation to serum creatinine in patients with a low cardiac index, there was no relation to the cardiac index. In patients with increases in PuO2 from day 1 to day 2, the cardiac index increased, and the serum creatinine level decreased on the 2nd day, whereas a sustained decrease in cardiac index and an increase in serum creatinine were observed in patients with a decrease in PuO2 from day 1 to day 2. Thus, PuO2 can be used as an indicator of the renal function in patients with ischemic heart disease. PMID- 9397310 TI - Pericardial effusion after streptokinase for acute myocardial infarction: an echocardiographic 1-year follow-up study. AB - Since the reported incidence of pericardial effusion following thrombolysis is highly variable, we have evaluated 80 consecutive patients with first acute myocardial infarction treated with streptokinase. Two-dimensional echocardiographic studies were performed on days 1, 2, 3, and 7, at 3 and 6 weeks, and 3, 6, and 12 months following acute myocardial infarction. Throughout the study, pericardial effusion was found in 7 of 80 (8.75%) patients, being small in 5 patients, moderate in 1, and large in 1 patient. No clinical, angiographic, or echocardiographic variable was associated with pericardial effusion formation. The incidence of pericardial effusion found in our study is almost three times lower than in other echocardiographic studies on pericardial effusion in thrombolysed patients. Whether this differences results from the beneficial effects of streptokinase is not clear. PMID- 9397311 TI - Early administration of ramipril in acute myocardial infarction: neurohormonal and hemodynamic effects and tolerability. AB - Although several large studies indicate a beneficial effect of angiotensin converting enzyme (ACE) inhibitors after myocardial infarction, the optimal timing of therapy in terms of safety and the effects on neurohormones during myocardial infarction are less well known. In order to investigate the effect of ramipril, administered within 24 h after myocardial infarction, on hemodynamics and neurohormones and its safety, 20 patients with a myocardial infarction were studied. Nine patients had an anterior, 10 an inferior, and 1 a non-Q-wave infarction. Fourteen patients received thrombolytic therapy, whereas 6 did not. The initial dose of ramipril was 1.25 mg, but was gradually increased to 5 mg during the next 4 days. Side effects did not occur. The mean arterial pressure decreased 8 h after the first dose from 84 +/- 2 mm Hg (control) to 77 +/- 2 mm Hg (p < 0.05) and remained decreased thereafter. This was accompanied by a reduction in systemic resistance of 8% after 8 h and of 12% on day 2. Heart rate, cardiac and stroke indexes, and pulmonary artery and wedge pressures did not change. The ACE activity decreased within 1 h of ramipril administration with a maximum of 71% at 4 h after the second dose and remained at this level throughout the study. Angiotensin II decreased by 34% (day 2) and by 41% (day 5). The renin activity gradually increased from 33 +/- 7.5 to 75.4 +/- 11.5 microM/ml on day 5, whereas epinephrine was reduced from day 2 onwards, with a maximal reduction of 71% on day 5. Arginine vasopressin was significantly reduced 5 h after ramipril administration until the end of the study, with a maximum of 77% on day 3. Moreover, a late but significant decrease in norepinephrine occurred on day 5. Thus, oral ramipril results in early ACE inhibition, followed by progressive attenuation of the neuroendocrine activation and a reduction in afterload during the acute phase of myocardial infarction. It is well tolerated, also in combination with nitroglycerin and thrombolytic therapy. PMID- 9397312 TI - Simultaneous dobutamine stress echocardiography and thallium-201 perfusion imaging for the detection of coronary artery disease. AB - To compare the diagnostic value of dobutamine stress echocardiography with dobutamine thallium-201 single-photon-emission computed tomography (SPECT) in detecting coronary artery disease, we performed both tests simultaneously on 93 patients who also underwent coronary arteriography. Dobutamine was infused at rates of 5, 10, 20, 30 and 40 microns/kg/min in 3-min stages. The left ventricle was divided into anteroseptal, posterolateral and inferior regions. Within each region, wall motion or perfusion abnormalities were classified as normal, ischemia or fixed defect. The response to stress was concordantly classified by both tests in 67 patients (72%, kappa = 0.48). Regional agreement for abnormalities was observed in 79% (kappa = 0.56) of the 279 regions analyzed. Dobutamine echocardiography detected 62 (93%) and thallium SPECT 60 (90%, p = NS) of the 67 patients with significant coronary artery disease (> or = 50% diameter stenosis). The specificity was 77 (20 of 26) and 81% (21 of 26), respectively. The accuracy was 88 and 87%, respectively. Combined the two tests gave a sensitivity of 97%, a specificity of 65% and an accuracy of 88%. The accuracy for detecting individual coronary stenosis with echocardiography was 83% for the left anterior descending artery, 84% for the right coronary artery and 73% for the left circumflex artery. With SPECT, it was 83, 87 and and 76%, respectively. In conclusion, dobutamine stress echocardiography and thallium SPECT provide a comparable accuracy for detection and localization of coronary artery disease, and for identification of regional myocardial abnormalities. Performing the two tests simultaneous is feasible but it adds limited value in detecting coronary artery disease. PMID- 9397313 TI - Influence of left ventricular diastole on left atrial appendage blood flow in patients with nonrheumatic atrial fibrillation. AB - The function of the left atrial appendage (LAA) reflected by the Doppler flow velocity at the outlet of the left atrial appendage has been reported to be correlated with spontaneous echo contrast and thrombus formation. To evaluate the influence of left ventricular diastole on LAA flow during atrial fibrillation (AF), 81 patients with chronic nonrheumatic AF were studied by transesophageal echocardiography. The peak outflow velocity of LAA during ventricular diastole was higher than that during ventricular systole (0.23 +/- 0.14 vs. 0.15 +/- 0.13 m/s, p < 0.001). The peak inflow velocity of LAA during ventricular diastole was also higher than that during ventricular systole (0.22 +/- 0.15 vs. 0.18 +/- 0.11 m/s, p < 0.01). Patients with a good left ventricular ejection fraction have a significantly higher peak LAA outflow velocity and a larger diastolic augmentation of LAA outflow (defined by the difference of LAA outflow between systolic and diastolic phases) than patients with an impaired left ventricular function. Thus, left ventricular diastole might have an influence on LAA flow during AF. In addition, the left ventricular function might be considered a predictor of subsequent thromboembolism from the viewpoint of its effect on LAA flow in patients with nonrheumatic AF. PMID- 9397314 TI - Risk stratification prior to vascular surgery: does the location of a dipyridamole thallium scintigram defect provide prognostic information? AB - BACKGROUND: While the value of myocardial scintigraphy using dipyridamole thallium is accepted for risk assessment prior to vascular surgery, it is unknown whether the location of the thallium abnormalities provides prognostic information. METHODS: Records from 435 consecutive patients scheduled for vascular surgery were reviewed and patients with dipyridamole thallium abnormalities involving the anterior distribution (ANTERIOR n = 62), or inferior or inferolateral distribution (INFERIOR n = 105) were assessed for cardiac complications of surgery (death, myocardial infarction, unstable angina or ischemic congestive heart failure). RESULTS: Patients with a normal dipyridamole thallium scintigraphy had few surgical cardiac complications: 2/86 (2%). Patients with an ANTERIOR dipyridamole thallium defect had a 12% incidence of surgical cardiac complications (7/57) without any cardiac deaths, while patients with an INFERIOR dipyridamole thallium defect had a similar incidence of surgical cardiac complications, 18% (18/100; p = 0.65 vs. ANTERIOR) including 4 cardiac deaths. CONCLUSIONS: Significant inferior or inferolateral dipyridamole thallium scintigraphy abnormalities are not associated with a lower risk of cardiac complications following vascular surgery than anterior abnormalities. Rather, any clearly abnormal dipyridamole thallium study is a marker for increased risk of perioperative events and may warrant further evaluation and treatment. PMID- 9397315 TI - On-line computerized vectorcardiography: influence of body position, heart rate, radiographic contrast fluid and myocardial ischemia. AB - On-line computerized vectorcardiography (cVCG) is increasingly being used for continuous monitoring of myocardial ischemia, however, little is known about factors other than ischemia causing electrocardiographic abnormalities. This paper describes how three important cVCG parameters, STC-VM, ST-VM and QRS-VD, are affected by different body positions, myocardial ischemia, contrast injection and increasing heart rate in patients with and without coronary artery disease. The main findings of the study are: contrast injection and different body positions caused major changes in QRS-VD but affected ST-VM and STC-VM to a minor degree. Increasing heart rate by atrial pacing produced substantial changes in all three parameters. Ischemia during angioplasty also produced changes in all three parameters, STC-VM being the most sensitive parameter. IN CONCLUSION: (1) STC-VM (> or = 50 microV) is the most valuable parameter for monitoring ischemia; (2) we propose ST-VM > or = 50 microV as criterion instead of previously used 25 microV; (3) QRS-VD cannot be used as a single marker of ischemia, and (4) electrocardiographic changes induced by increased heart rate should be taken into account during interpretation. PMID- 9397316 TI - PVF velocity pattern in patients with heart failure: transesophageal echocardiographic assessment. AB - In order to assess the role of the pulmonary venous flow (PVF) velocity pattern in the evaluation of patients with congestive heart failure (CHF), we studied 41 CHF patients by means of transthoracic echocardiography (TTE) and multiplane transesophageal echocardiography (TEE). The etiology of CHF was idiopathic or ischemic dilated cardiomyopathy in 19 patients and hypertensive heart disease in 22. Sixteen subjects without cardiovascular disease were selected as normal controls. PVF peak systolic and peak early diastolic (D) velocities were recorded by TEE and TTE and the systolic fraction (SF) was measured (i.e., the systolic velocity-time integral-VTI-expressed as a fraction of the sum of systolic and early diastolic (VTI). TEE tracings were obtained in all patients and had more laminar-appearing spectral signals, thus were used for analysis. By TEE the mitral flow velocity patterns were also evaluated: peak early diastolic velocity (E), peak velocity at atrial contraction, E velocity normalized for VTI (E/VTI), deceleration time (DT), and left ventricular isovolumic relaxation time (LVIRT). The left ventricular ejection fraction (LVEF) was calculated by two-dimensional echocardiographic images using the modified Simpson method. The SF was lower in CHF patients as compared with normal controls (p < 0.0001). The E/VTI ratio was higher, and DT and LVIRT were shorter (p < 0.0001) in CHF patients. A significant correlation was observed between SF and LVEF in CHF patients (r = 0.76, p < 0.001). Two different PVF velocity patterns (type A:SF << 50%, D > 50 cm/s; type B:SF approximately 50%, D > 50 cm/s) were recognized in patients with a low LVEF (type A) and a nearly normal or normal LVEF (type B). Patients with LVEF < 40% showed mean SF values significantly lower than patients with LVEF > 40% (33.26 +/ 10.84 vs. 51.00 +/- 4.00%, p < 0.0001). Mean DT and LVIRT values were not significantly different in patients with LVEF < 40% and > 40%. Thus in CHF patients TEE PVF velocity patterns help in distinguishing patients with systolic dysfunction (low LVEF and SF) from patients with predominant diastolic impairment (normal or nearly normal LVEF, high D velocities). PMID- 9397317 TI - Effect of controlled exercise training in coronary artery disease patients with and without left ventricular dysfunction assessed by cardiopulmonary indices. AB - Cardiopulmonary indices were used to evaluate the effect of controlled exercise training prescribed on the basis of the heart rate at the ventilatory anaerobic threshold in coronary artery disease patients with and without impaired left ventricular function. Fifty-two patients aged 38-75 years were divided into four groups. The first three groups included patients with a left ventricular ejection fraction of > 45% at rest, as follows: group 1, 10 patients with single-vessel disease; group 2, 12 patients with two-vessel disease; group 3, 10 patients with three-vessel disease. Group 4 comprised 20 patients with left ventricular dysfunction (ejection fraction < 35%). The left ventricular ejection fraction was assessed by multigated acquisition radionuclear study. All patients underwent a cardiopulmonary exercise test before and after the program which lasted 6-9 months. The variables measured were oxygen consumption (VO2), CO2 output, minute ventilation, O2 pulse, and ventilatory anaerobic threshold. Significant improvements in maximal VO2, maximal O2 pulse, and ventilatory anaerobic threshold level were observed in groups 1, 2, and 4 (p < 0.1-0.0001), but not in group 3. These findings indicate that the overall cardiac function, as evaluated by cardiopulmonary indices, improves in patients with one- or two-vessel disease with good left ventricular function and in patients with impaired left ventricular function following an exercise training program. Severe coronary disease seems to limit improvement, even in the presence of a good left ventricular function. The results validate the heart rate at the ventilatory anaerobic threshold as the optimal training heart rate in coronary artery disease patients and the cardiopulmonary exercise test as a sensitive tool for evaluating exercise training results. PMID- 9397318 TI - Spongy myocardium. AB - Spongy myocardium is a rare congenital anomaly. We report a 35-year-old patient in whom diagnosis of spongy myocardium had been made by angiocardiography 20 years before. The disorder eventually resulted in progressive right and left heart failure. PMID- 9397319 TI - Anomalous origin of all coronary arteries from the pulmonary trunk. AB - The origin of both coronary arteries from the pulmonary artery is a rare cardiac malformation. We report a baby who presented with an echocardiographically diagnosed perimembranous ventricular septal defect and normal left ventricular (LV) function. Later on the boy developed failure to thrive and increasing tachypnea. At the age of 5 weeks the ECG showed that LV strain and echocardiographic LV function had worsened (FS 18%). Echocardiography and heart catheterization showed that all coronary arteries originated from the pulmonary trunk. Intraoperative inspection revealed a single ostium for the right and left coronary artery in the nonfacing sinus of the pulmonary trunk. A tube was constructed connecting the coronary artery to the ascending aorta. Coronary perfusion was sufficient and the sinus rhythm was restored. However, in the early postoperative period there was a sudden deterioration of cardiac output followed by cardiac arrest. Reanimation was not successful. PMID- 9397320 TI - Viable but denervated right ventricular myocardium: a case of Eisenmenger reaction. AB - We report on a 47-year-old woman who experienced an Eisenmenger reaction (mean pulmonary artery pressure: 86 mm Hg) induced by atrial septal defect. Radionuclide myocardial scans with 99mTc-MIBI and 123I-BMIPP showed increased uptake to the right ventricular myocardium, whereas the 123I-MIBG scan disclosed no uptake to the right ventricular myocardium. The scans showed no apparent abnormality in the left ventricular myocardium. These findings suggest that the right ventricular myocardium was viable but denervated due to severe pulmonary hypertension. The mechanism of right ventricular failure may be closely related to sympathetic denervation. PMID- 9397321 TI - Primary cardiac lymphoma with infiltration of the atrioventricular node: remission with reversal of the atrioventricular block induced by chemotherapy. AB - We describe a rare case of primary cardiac lymphoma associated with complete atrioventricular block, with recovery following systemic chemotherapy. A 61 year old woman was admitted because of a massive pericardial effusion and complete atrioventricular block. Echocardiography disclosed several polypoid masses in the right atrium. A diagnosis of primary cardiac lymphoma was made. After three cycles of chemotherapy, the patient achieved a complete remission, the ECG showed first-degree atrioventricular block, and echocardiography revealed disappearance of the right atrial masses. This is the first report of a patient recovering from primary cardiac lymphoma and complete atrioventricular block following systemic chemotherapy. We conclude that aggressive treatment of this malignancy can improve cardiac function and the patient's prognosis. PMID- 9397322 TI - Interaction between cortisol and tumour necrosis factor with concurrent resistance and endurance training. AB - OBJECTIVE: To determine the effect of concurrent resistance and endurance training on tumor necrosis factor alpha (TNF alpha), urinary free cortisol, strength [one-repetition maximum (1 RM)], and maximal oxygen consumption (Vo2max). DESIGN: Randomized control trial of 12 weeks' duration. SETTING: University of Alberta, Edmonton, Alberta, Canada. PARTICIPANTS: Forty-five healthy female (n = 18) and male (n = 27) subjects who had not formally trained for at least 6 months prior to the study but were physically active. The mean +/- SD age, height, and body mass for all subjects were 22.3 +/- 3.3 years, 1.76 +/- 9.32 m, and 73.4 +/- 11.6 kg, respectively. INTERVENTION: The subjects were randomly assigned to four groups: strength training only (S), n = 10; endurance training only (E), n = 11; combined strength and endurance training (SE), n = 13; and a control group (C), n = 10. The S and E groups performed progressively overloaded training sessions three times per week for 12 weeks. The SE group completed the same strength and endurance training programs on different days (i.e., 6 days/week) for 12 weeks. MAIN OUTCOME MEASURES: Serum levels of TNF alpha, urinary free cortisol, 1 RM, and Vo2max were measured before and after 6 and 12 weeks of training. RESULTS: Significant increases in leg press and knee extension 1 RM occurred after training in both S and SE groups, but the relative gains in knee extension 1 RM were greater in the S group. Similar increases in Vo2max were observed in groups E and SE (p < 0.05). Cortisol was significantly increased in the SE group for women and decreased in the E group for men after training. TNF alpha was significantly elevated in the women of group E after training. No correlation was observed between urinary free cortisol and TNF alpha with training. CONCLUSION: These results indicate that a partial interference effect of compromised strength gains in unilateral knee extension of the men occurred after concurrent strength and endurance training that could not be attributed to an interaction between cortisol and TNF alpha in response to this type of exercise. PMID- 9397323 TI - Injuries of young elite female basketball players over a six-year period. AB - OBJECTIVE: To analyze injuries retrospectively among female basketball players at the Australian Institute of Sport (AIS) from 1990 to 1995 inclusive. DESIGN: The medical records of all the female basketball players on AIS (residential) scholarships were examined, and all injuries were recorded. SETTING: The Sports Medicine Department at the Australian Institute of Sport in Canberra, Australia. PARTICIPANTS: The participants were 49 elite female basketball players, holding full scholarships at the AIS, with an average age of 17.6 years at the time of injury presentation. MAIN OUTCOME MEASURES: Injury presentation according to region involved, nature of injury, and most common specific injuries (diagnoses). RESULTS: A total of 223 injuries were recorded: 139 were acute and 84 were chronic. The regions most frequently injured were the knee (18.8%), ankle (16.6%), lumbar spine (11.7%), and lower legs (10.8%). The most frequent diagnoses were ankle lateral ligament sprain (12.1%), patellar tendinitis (6.7%), lower limb stress fractures (5.4%), finger sprains (4.9%), and mechanical low back pain (4.5%). CONCLUSIONS: There was a high incidence of knee and ankle injury in this group of young elite female basketball players, and stress fractures were not uncommon. The incidence of injury in female basketball players may be increasing. Further research in this area may help reduce the risk of stress fractures and serious ankle and knee injuries. PMID- 9397324 TI - Spirometry and airway reactivity in elite track and field athletes. AB - OBJECTIVES: To characterize spirometry and to document the incidence of exercise induced bronchospasm (EIB) during competition in elite track and field athletes. DESIGN: Spirometry was performed in 120 men and 69 women athletes before competition and peak expiratory flows in 50 men and 23 women athletes before and after competition. SETTING: The 1991 (Randalls Island, NY, U.S.A.) and the 1993 (Eugene, OR, U.S.A.) National Track and Field Championships (World Championship team-qualifying meet). PARTICIPANTS: American track and field athletes who met World Championship qualifying standards. MEASUREMENTS: Spirometry (Cybermedic, Inc., Boulder, CO, U.S.A.) and peak expiratory flows (Personal Best, Healthscan Products, Cedar Grove, NJ, U.S.A.)--the best of three reproducible efforts. RESULTS: Male sprinters had lower vital capacities than other track athletes, whereas both male and female field (throwing) athletes had larger vital capacities than both runners and other field athletes. Decreases of 10% peak expiratory flows were found in 10% of men and 26% of women track athletes within 15 min after competition. The incidence was higher in longer-distance events. Most participants did not have a history of asthma. CONCLUSIONS: A higher-than expected prevalence of EIB was found in high-level track athletes. The results suggest that spirometry and/or peak flows should be measured in track athletes because small decreases in airflow may impair training or performance, a condition that is easily treated. PMID- 9397325 TI - Lifestyles and health risks of collegiate athletes: a multi-center study. AB - OBJECTIVE: To determine whether college athletes are at greater risk for maladaptive lifestyle and health-risk behaviors than their nonathletic peers and to identify high risk taking groups by gender, sport, and other identifiers. DESIGN: Multicenter, cross-sectional study. SETTING: Seven major geographically represented collegiate institutions in the United States. PARTICIPANTS: A total of 2,298 college athletes and 683 randomized nonathlete controls completed a confidential survey questionnaire between the summer of 1993 and winter of 1994, assessing lifestyle and health-risk behaviors over the previous 12 months. MAIN OUTCOME MEASURES: Self-reports of lifestyle behaviors and health risks in the following areas: motor-vehicle safety, substance abuse, sexually transmitted diseases and contraception, mental health, cancer prevention, nutrition, exercise and general preventive health issues. RESULTS: Athletes demonstrated significantly higher risk-taking behaviors (p < 0.05) than their nonathlete peers in the following areas: less likely always to use seatbelts; less likely always to use helmets with motorcycles, mopeds, and bicycles; more often drive as a passenger with a driver under the influence of alcohol or drugs; greater quantity and frequency of alcoholic beverages; greater frequency of smokeless tobacco and anabolic steroid use; less-safe sex; greater number of sexual partners; less contraceptive use; and more involvement in physical fights. Female athletes reported a higher prevalence of irregular menses, amenorrhea, and stress fractures compared with female nonathletes. Male athletes had more risk-taking behaviors than did female athletes (p < 0.05), and athletes in contact sports demonstrated more risk-taking behaviors than did athletes in noncontact sports (p < 0.05). Athletes with one risk-taking behavior were likely to have multiple risk taking behaviors (p < 0.05). CONCLUSIONS: College athletes appear to be at higher risk than their nonathletic peers for certain maladaptive lifestyle behaviors. Athlete subgroups at highest risk include male athletes and athletes participating in contact sports. Athletes at risk for one high-risk behavior demonstrated an increased risk for multiple risk-taking behaviors. Preventive health interventions deserve further study to determine strategies for risk reduction in high-risk groups. PMID- 9397326 TI - Management guidelines for participation in collision activities with congenital, developmental, or postinjury lesions involving the cervical spine. AB - OBJECTIVE: Conditions involving the cervical spine in athletes requiring a management decision are numerous. This report presents appropriate guidelines for return to collision activities in those with congenital, developmental, or postinjury lesions. DATA SOURCES: Information was compiled from > 1,200 cervical spine lesions documented by the National Football Head and Neck Injury Registry and an extensive literature review. DATA SYNTHESIS: Available data as well as a clinical understanding of injury mechanisms have resulted in the development of reasonable management guidelines. Each of the congenital, developmental, and posttraumatic conditions presented are determined to present either no contraindication, relative contraindication, or an absolute contraindication to sport participation on the basis of a variety of parameters. Conditions included in the discussion are odontoid anomalies; spina bifida occulta; atlanto-occipital fusion; Klippel-Feil anomalies; cervical canal stenosis; spear tackler's spine; traumatic conditions of the upper, middle, and lower cervical spine, including ligamentous injuries and fractures; intervertebral disc injuries; and postcervical spine fusion. CONCLUSION: The proposed guidelines should be used in the decision-making process in conjunction with other such factors as the age, experience, ability of the individual, level of participation, and position played, as well as the attitude and desires of the athlete and his or her parents following an informed discussion of the problem with particular regard to potential risk. PMID- 9397327 TI - Collapsed ultraendurance athlete: proposed mechanisms and an approach to management. AB - OBJECTIVE: To review the common conditions causing collapse in ultraendurance athletes, to propose appropriate treatment protocols for the more common medical conditions that are encountered, and to provide practical guidelines for the management of the medical facilities at these endurance events. DATA SOURCES: Books published on the subject, abstracts of the American College of Sports Medicine Annual Congress dealing with the subject over the last 5 years, and electronic search of the Medline and Current Contents databases (last searched May 1995). German articles were included in the search criteria. STUDY SELECTION: Articles dealing generally with the management of medical facilities at endurance events were chosen. Articles dealing more specifically with the common medical conditions encountered at these events were then sourced and included in this review. DATA EXTRACTION: Multiple reader extraction of relevant data. DATA SYNTHESIS: A practical guideline to the management of the medical facility at an endurance event is detailed, expanding specifically on the nature of the conditions causing collapse during or after endurance exercise with a proposed classification of causes of collapse for optimizing immediate management and management protocols for specific conditions such as heatstroke, hyponatremia, hypoglycemia, exercise-associated collapse, and muscle cramps. CONCLUSION: The most commonly encountered medical condition at endurance athletic events is collapse after the event. The popular view that all persons who collapse have dehydration-induced hyperthermia has been challenged. Here we extend that argument and provide diagnostic and management protocols and propose a triage system that considers the most common causes of collapse in ultraendurance athletes. These protocols can optimize the efficient and safe management of large numbers of collapsed ultraendurance athletes. It is proposed that these protocols can optimize the efficient and safe management of large numbers of collapsed ultra endurance athletes. PMID- 9397328 TI - Does repetitive physical loading inhibit radial growth in female gymnasts? AB - OBJECTIVE: Stress-related injuries to the distal radius have been noted in female gymnasts with potential for resultant premature closure and abnormal growth at this site. The purpose of this study was comprehensively to review and critically to appraise the available literature to examine the evidence related to this question: does repetitive physical loading inhibit growth of the radius in female gymnasts? DATA SOURCES: MEDLINE and SPORT Discuss were searched from 1975 to the present by using "gymnast" in combination with injury, growth plate, epiphyseal, and ulnar variance. Additional references were retrieved from the bibliographies of the retrieved articles. STUDY SELECTION: All descriptive and analytic studies that included data related to stress-related injuries affecting the distal radius of competitive female gymnasts were included. Conclusions regarding the effects of gymnastics training on radial growth of female gymnasts were limited to data from case reports, clinical series, cross-sectional studies, and descriptive cohort studies. Data from relevant experimental animal studies also were included. DATA EXTRACTION AND SYNTHESIS: In reviewing the literature, particular attention was paid to the relative strengths of the different study designs. From these data, information associated with growth inhibition at the distal radius was examined. MAIN RESULTS: The descriptive research reviewed included clinical, cross-sectional, and cohort studies that establish the existence of stress related injuries affecting one or more constituent parts of the epiphyseal physeal-metaphyseal (EPM) complex of the distal radius, symptomatic ulna-radial length difference (URLD), and distal radius physeal arrest among female gymnasts. Five cross-sectional studies showed radiographic abnormalities consistent with distal radius physeal-stress reaction in 10-85% of gymnasts studied. Two cross sectional studies indicated "abnormal" positive URLD in 8-20% of wrists radiographed. Four cross-sectional studies showed significant correlations between training intensity and URLD, suggesting a dose-response relation. Three cross-sectional studies indicate greater URLD in gymnasts compared with nongymnasts. Radiographic evidence of distal radius physeal arrest involving physically immature female gymnasts is presented in four studies (two clinical series, one cross-sectional, and one descriptive cohort). In animal studies, prolonged physical training has also been shown to inhibit or stop growth in weight-bearing long bones. However, there were no rigorous studies (i.e., randomized control trials or analytic cohorts) examining the question. CONCLUSION: The results of this critical review of the scientific literature support the plausibility of stress-related distal radius physeal arrest with secondary URLD. However, the strength of evidence is inadequate to be conclusive. PMID- 9397329 TI - Pulmonary contusion secondary to blunt trauma in a collegiate football player. PMID- 9397330 TI - Putting principles into practice: the evolution of evidence-based medicine. PMID- 9397331 TI - Assessment of childhood phobias. AB - Childhood phobias can be successfully treated using a variety of behavioral strategies, provided there has been a psychometrically sound assessment. Measures are also important for the evaluation of treatment efficacy and the testing of hypotheses generated by new ideas and theories of children's phobias. This paper outlines broad-based assessment procedures used in the evaluation of children's phobias, including the behavioral or problem-focused interview, the diagnostic interview, self-report inventories, caregiver completed instruments, behavioral observations, self-monitoring and physiological assessment. Reflecting recent theoretical and clinical advances in the study of childhood internalizing disorders, we also explore laboratory-based measures and family assessment measures. Particular attention is given to psychometric issues and developmental sensitivity in our discussion of these assessment procedures. PMID- 9397332 TI - Treatment approaches for pathological gamblers. AB - Outcome literature on the treatment of pathological gambling is reviewed, encompassing psychodynamic, behavioral, cognitive, cognitive-behavioral, multimodal, pharmacotherapeutic, and 12-step approaches. No properly controlled research has been conducted with psychodynamic or 12-step methods, and pharmacotherapies require replications with larger samples to determine their efficacy. Multimodal approaches have been tested most often in inpatient settings, and given the range of methods combined it is difficult to infer specific efficacy for treatment components. The largest volume of outcome research has focused on behavioral, cognitive, and combined cognitive-behavioral treatment methods, and findings from controlled and uncontrolled trials provide support for efficacy of these approaches. As a whole, the literature indicates that pathological gambling can be treated with highly successful outcomes. Needs for further research are considered. PMID- 9397333 TI - Meta-analysis of cognitive-behavioral treatment studies for bulimia. AB - A meta-analysis was performed to systematically assess the effect of cognitive behavioral treatments for bulimia. To protect against past criticisms of meta analyses, this study focused on well-defined hypotheses with clearly articulated conceptual foundations. Twenty-six studies of the cognitive-behavioral treatment of bulimia were selected through computer searches. Effect sizes were calculated for changes in behavioral outcome measures (25 independent hypothesis tests) and cognitive-attitudinal outcome measures (17 independent hypothesis tests). Additionally, two effect sizes were generated for within and between group comparisons. The analysis revealed an effect size of average r = 0.69 for behavioral outcome measures (average r = 0.64 for between group and average r = 0.74 for within group) and average r = 0.67 for cognitive-attitudinal outcome measures (average r = 0.64 for between group and average r = 0.69 for within group). Follow-up effect sizes were less favorable; however, the diversity of time spans and outcome measures used to calculate follow-up effect sizes limit their utility. Overall, results suggest that the use of a cognitive-behavioral therapy will result in favorable treatment outcomes and implications for future research are discussed. PMID- 9397334 TI - A biopsychosocial model of metaphor therapy with holistic cultures. AB - For centuries western cultures have adopted a dualistic perspective toward people's health. The "self" has emerged as an independent entity from others as well from the body. Human distress has been psychologized and depression and anxiety have been attributed to intrapsychic structures and processes. Nevertheless, many nonwestern cultures still adopt holistic perspectives. Within these cultures, distress is manifested through physical rather than psychological complains. Therefore, psychological approaches, based on the independence of the self, may not be fitting for these societies. Instead, based on the assumption that nonwestern cultures are holistic and less psychologized and their problems are social rather than intrapsychic, a biopsychosocial approach is suggested. In addition, nonwesterners have a different concept of reality. For instance, within some communities fantasies and delusions are appreciated, constitute part of a normal life, and are considered to be the "real reality." Furthermore, complains are often described in metaphoric language. Accordingly, a biopsychosocial model of metaphoric therapy is proposed in which therapists would incorporate their clients' metaphoric imaginative culture. Metaphoric intervention also allow for changes in the biological, psychological, and sociocultural reality of the client. PMID- 9397335 TI - "The myth of mental illness:" continuing controversies and their implications for mental health professionals. AB - Since the publication of "The Myth of Mental Illness" in 1960, there has been an ongoing debate about Thomas Szasz's ideas concerning mental illness. In this paper, Szasz's views are summarized, as are the views of Szasz's critics. Specifically, the following areas are addressed: Szasz's definition of disease, his notions regarding the unconscious and rationality, his beliefs regarding culpability, his proposed differences between psychiatry and other branches of medicine, the uses of the term "mental illness," and the possibility of implicating physical lesions in some mental illnesses. With this discussion as a backdrop, the importance of these issues to mental health practitioners is addressed. PMID- 9397336 TI - Acute stress disorder: a critical review of diagnostic issues. AB - Acute stress disorder (ASD) is a recently developed diagnosis that describes posttraumatic stress reactions that occur in the first month following a trauma. Diagnostic criteria include dissociative, intrusive, avoidance, and arousal symptoms. ASD was driven by the proposal that trauma leads to dissociative reactions, and these are predictive of longer-term psychopathology. This paper reviews a series of anomalies in the diagnostic criteria, highlights discrepancies between criteria for ASD and posttraumatic stress disorder (PTSD), and illustrates the lack of empirical evidence for some assumptions inherent in the conceptualization of ASD. It is argued that future revisions of ASD criteria need to be based on empirical evidence of acutely traumatized individuals. PMID- 9397337 TI - Staff beliefs about the challenging behaviors of children and adults with mental retardation. AB - From both theoretical and practical perspectives, staff beliefs are likely to have a significant impact on the process of care for children and adults with mental retardation who engage in challenging behaviors. This paper reviews research addressing three domains of staff beliefs: definitions of challenging behavior, causal attributions, and beliefs about appropriate intervention. In general, staff definitions were found to be at odds with formal definitions. According to care staff, challenging behaviors are actions that are difficult to manage. Staff causal attributions appear congruent with current theory, when measured with little specificity. However, when staff are asked to suggest causes of challenging behavior with clearly described functions they often fail to make appropriate attributions. Beliefs about appropriate short-term interventions suggest responses likely to develop and/or maintain challenging behavior, but beliefs about longer-term planned intervention appear to be more closely matched to contemporary practice. Reasons for this long-term/short-term distinction, based on the demands of the immediate situation, are proposed. Suggestions for future research on staff beliefs are discussed in detail. Finally, implications for staff training, referral practice, and for analysis and intervention with challenging behavior, are outlined. PMID- 9397338 TI - Individual differences in the experience of emotions. AB - The purpose of this paper is to highlight the important role of individual difference factors in the experience of emotion. We begin by describing several commonalties across two major approaches to the study of emotion, namely, the neuropsychological and cognitive perspectives. Both approaches provide some degree of support for the role of individual differences and cognitive factors in the experience of emotion. This paper builds on these commonalities by reviewing personality and psychopathology findings that indicate the contribution of both positive and negative personality characteristics (e.g., extraversion, optimism vs. neuroticism, trait anxiety) to the types of cognitive appraisals and emotional responses exhibited by different individuals. A self-schema model of emotion is presented as a means of integrating more fully this individual differences perspective with a theory of emotion. In this model, self-schema content provides the basis for individual differences in underlying core themes and self-evaluative beliefs. The model describes how self-schema content distinctions across individuals may have a differential impact on the initial processing of an event, evaluation of this event with respect to the self, and emotional and behavioral output. Several examples are then presented to illustrate the increased predictability afforded by this individual differences based self-schema model of emotion. The application of this model to treatment and prevention issues in clinical and health psychology is also briefly considered. Finally, the model is integrated with other theoretical perspectives on emotion by describing a number of additional research and theoretical implications. Emphasis is placed on the need for further clarification of both cognitive and emotional components of an individual differences perspective on the study of emotions. PMID- 9397339 TI - Temporal reasoning and temporal data maintenance in medicine: issues and challenges. AB - We present a brief, nonexhaustive overview of research efforts in designing and developing time-oriented systems in medicine. The growing volume of research on time-oriented systems in medicine can be viewed from either an application point of view, focusing on different generic tasks (e.g. diagnosis) and clinical areas (e.g. cardiology), or from a methodological point of view, distinguishing between different theoretical approaches. In this overview, we focus on highlighting methodological and theoretical choices, and conclude with suggestions for new research directions. Two main research directions can be noted: temporal reasoning, which supports various temporal inference tasks (e.g. temporal abstraction, time-oriented decision support, forecasting, data validation), and temporal data maintenance, which deals with storage and retrieval of data that have heterogeneous temporal dimensions. Efforts common to both research areas include the modeling of time, of temporal entities, and of temporal queries. We suggest that tasks such as abstraction of time-oriented data and the handling of different temporal-granularity levels should provide common ground for collaboration between the two research directions and fruitful areas for future research. PMID- 9397340 TI - Modeling medical trials in pharmacoeconomics using a temporal object model. AB - Time is an inherent feature of many medical applications. These applications can also benefit from the support of object database management systems which better capture the semantics of the complex objects that arise in the medical domain. In this paper, we present a uniform behavioral temporal object model which includes a rich and extensible set of types and behaviors to support the various features of a medical application. We concentrate here on the application of pharmacoeconomic medical trials. Pharmacoeconomics is a field of medical economics in which the costs and outcomes of alternative treatments are assessed and compared, in order to establish which is the most appropriate treatment for a particular illness in a particular setting. We describe in detail the histories and timelines features of our temporal model and show how they can effectively be used to model a pharmacoeconomic trial. We then give an instance of a pharmacoeconomic trial as it would appear in the temporal object model and show, using queries, how a series of different behaviors could be used to retrieve various components of the instance. These components could then be used to assess the alternative treatments involved in the trial and determine their cost effectiveness. PMID- 9397341 TI - Effective data validation of high-frequency data: time-point-, time-interval-, and trend-based methods. AB - Real-time systems for monitoring and therapy planning, which receive their data from on-line monitoring equipment and computer-based patient records, require reliable data. Data validation has to utilize and combine a set of fast methods to detect, eliminate, and repair faulty data, which may lead to life-threatening conclusions. The strength of data validation results from the combination of numerical and knowledge-based methods applied to both continuously-assessed high frequency data and discontinuously-assessed data. Dealing with high-frequency data, examining single measurements is not sufficient. It is essential to take into account the behavior of parameters over time. We present time-point-, time interval-, and trend-based methods for validation and repair. These are complemented by time-independent methods for determining an overall reliability of measurements. The data validation benefits from the temporal data-abstraction process, which provides automatically derived qualitative values and patterns. The temporal abstraction is oriented on a context-sensitive and expectation guided principle. Additional knowledge derived from domain experts forms an essential part for all of these methods. The methods are applied in the field of artificial ventilation of newborn infants. Examples from the real-time monitoring and therapy-planning system VIE-VENT illustrate the usefulness and effectiveness of the methods. PMID- 9397342 TI - Representing and developing temporally abstracted knowledge as a means towards facilitating time modeling in medical decision-support systems. AB - The utilization of the appropriate level of temporal abstraction is an important aspect of time modeling. We discuss some aspects of the relation of temporal abstraction to important knowledge engineering parameters such as model correctness, ease of model specification, knowledge availability, query completeness, inference tractability, and semantic clarity. We propose that versatile and efficient time-modeling formalisms should encompass ways to represent and reason at more than one level of abstraction, and we discuss such a hybrid formalism. Although many research efforts have concentrated on the automation of specific temporal abstractions, much research needs to be done in understanding and developing provably optimal abstractions. We provide an initial framework for studying this problem in a manner that is independent of the particular problem domain and knowledge representation, and suggest several research challenges that appear worth pursuing. PMID- 9397343 TI - Using a general theory of time and change in patient monitoring: experiment and evaluation. AB - In this paper, we propose to use one of the well-known general theories of time and change, namely the Event Calculus (Kowalski and Sergot, New Generation Computing 4, 67-95, 1986), to represent temporal aspects in intelligent medical monitoring systems. In particular, we explore the application of CEC (Chittaro and Montanari, Computational Intelligence 12, 359-382, 1996) (an efficient implementation of the Event Calculus) to the management of mechanical ventilation. First, we present the prototype we have built, which has been extensively tested on patient's data from real clinical cases. Then, we provide a thorough evaluation of the obtained results, pointing out both strengths and weaknesses of the approach, and identifying a number of extensions which can be extremely useful to scale up the medical application of the approach. PMID- 9397344 TI - Efficient temporal probabilistic reasoning via context-sensitive model construction. AB - We present a language for representing context-sensitive temporal probabilistic knowledge. Context constraints allow inference to be focused on only the relevant portions of the probabilistic knowledge. We provide a declarative semantics for our language. We present a sound and complete algorithm for computing posterior probabilities of temporal queries, as well as an efficient implementation of the algorithm. Throughout we illustrate the approach with the problem of reasoning about the effects of medications and interventions on the state of a patient in cardiac arrest. We empirically evaluate the efficiency of our system by comparing its inference times on problems in this domain with those of standard Bayesian network representations of the problems. PMID- 9397345 TI - The prophenoloxidase activating system of the shrimp Penaeus paulensis and associated factors. AB - In the present study we investigated the proPO activating system of the penaeid Penaeus paulensis, focusing on its role in the shrimp immune system. The great majority of PO activity (more than 90%) was found in the shrimp hemocytes. The enzyme activity was greatly enhanced by components of microorganism cell walls, such as LPS and beta-1,3-glucans, suggesting its involvement in non-self recognition. PO activity was also found in the shrimp serum and trypsin, and LPS were able to increase the enzyme activity. Thus, serum can be used as an alternative for the study of the shrimp proPO activating system, as it is much more readily obtained than HLS. PO activity was cation-dependent, and 5 mM of calcium and 10 mM of magnesium were the optimal concentrations for the enzyme activity. An immune factor was found in the shrimp HLS, capable of inducing cell adhesion and degranulation of the penaeid hemocytes. PMID- 9397346 TI - Molecular cloning of double-stranded RNA inducible Mx genes from Atlantic salmon (Salmo salar L.). AB - Mx proteins are induced by type I interferons in mice and humans and inhibit the replication of several negative-stranded RNA viruses. In this work Mx genes in Atlantic salmon were studied using the double stranded RNA, polyinosinic: polycytidylic acid (poly I:C), to induce interferon production. Northern blot analysis showed Mx mRNA in liver, head kidney and gills 2 days after poly I:C injection of fish, but not in untreated fish or fish injected with saline or LPS. Mx transcripts of 2.4 and 1.9 kb were detected in the liver. By screening of a cDNA library, three different full length Mx cDNA clones, ASMx1, ASMx2 and ASMx3, were identified and sequenced. The deduced ASMx proteins all contain 623 amino acids and show the tripartite GTP-binding motif typical of vertebrate Mx proteins. ASMx1 and ASMx2 may represent alleles of the same gene whereas ASMx3 represents a different gene. The deduced ASMx proteins showed 96 to 98% sequence identity with rainbow trout Mx1 and Mx3 and about 88% identity with rainbow trout Mx2 protein. PMID- 9397347 TI - Identification of a differential display product associated with apoptosis in chicken thymocytes. AB - To further elucidate the cellular mechanisms that mediate programmed cell death in avian immune cells, differential display analysis was employed to identify differentially expressed genes in chicken thymocytes undergoing apoptosis. Primary cultures of thymocytes were treated with dexamethasone to activate apoptosis and RNA was isolated for differential display analysis. A differential display product designated A1 (479 bp) was identified. This display product was subcloned and induced expression of the genes was confirmed by ribonuclease protection analysis. Nucleotide sequence analysis of A1 revealed a putative 82 amino acid open reading frame that demonstrated limited homology with Bad, an apoptotic regulatory protein. Thus, A1 may represent the avian homolog of Bad. PMID- 9397348 TI - Phenotyping of beluga whale blood lymphocytes using monoclonal antibodies. AB - Widespread efforts are currently made to classify morphologically indistinguishable lymphocyte subpopulations in several species. In order to increase the knowledge in cetacean immunology, cross-reactivity of antibodies against bovine, human, ovine and mouse cell surface proteins was tested on beluga whale (Delphinapterus leucas) peripheral blood lymphocytes using flow cytometry. Anti-MHC class I and II as well as anti-CD2 reacted with virtually all peripheral blood lymphocytes. Anti-TCR gamma delta and anti-CD4 reacted with respectively 31% and 30% of peripheral blood lymphocytes. B lymphocytes were identified by an anti-surface IgM which was present on 6% of blood lymphocytes. Specificity of these antibodies was demonstrated by immunoprecipitation of beluga proteins with similar molecular weight to that of other species. These results could be useful for further immunotoxicological evaluation of highly versus mildly contaminated populations of belugas. PMID- 9397349 TI - T cell progenitors in the murine small intestine. AB - Lymphocytes in the murine small intestine epithelium are known to have a high proportion of extrathymic T cells. To explore the possibility that small intestine intraepithelial lymphocytes (IELs) are derived from T cell progenitors present within the intestine, intestine-derived cells with characteristics of early-stage T cell precursors were studied for their ability to regenerate IEL T cell populations following transfer into irradiated recipient mice. Cells within this population lacked markers of mature T cells but expressed heat-stable antigen, the c-kit receptor for stem cell factor, and/or the pre-T cell alpha gene. Upon adoptive transfer, donor cells preferentially homed to the intestine and did not repopulate the thymus or extraintestinal peripheral lymphoid tissues. IELs derived from the donor precursor pool included both (alpha beta and gamma delta T subsets and consisted of phenotypically heterogeneous cell populations defined by CD4 and CD8. These findings provide evidence that T cell progenitors located in the intestinal mucosa are the likely source of most intestinal IELs. PMID- 9397350 TI - Age-related changes of naive and memory CD4 rat lymphocyte subsets in mucosal and systemic lymphoid organs. AB - The purpose of this study was to investigate in rats, by double-label immunofluorescence and flow cytometric analysis, the age related changes in the CD4 subset of gut-associated lymphoid tissues and spleen. We found that the percentage of CD4+ T cells in Peyer's patches (PP) and spleen (SP) increased during the first 6 weeks after weaning. An age-related decrease of the CD4 subset was observed in SP of aged rats, but not in their PP. In all lymphoid tissues studied, an age-related decrease of the Thy-1+ subset was observed from weaning to 2 years of age. Analysis of the naive CD4 subset (CD45RC+) showed that in SP this subset increased during the first 9 weeks of age, and declined in aged rats. However, in PP this subset presented a slow decrease from weaning until 2 years of age. Together with the decrease of the naive subset, a sharp increase of the memory/activated CD4+ cells (CD45RC- Thy-1-) was observed in PP, and to a lesser extent in SP. When the maturation of the CD4 T cells in PP was followed during the first week after weaning, we found that an important proportion of this subset changes its phenotype at this time, from recent thymic emigrant (CD45RC- Thy-1+) to naive T cell (CD45RC+ Thy-1-) and then to activated/memory cell (CD45RC- Thy-1-). Therefore it appeared that CD4 T cells from PP mature faster than SP CD4 T cells, and they are not subject to the deleterious effect of aging. One surprising point was the different kinetics of the CD4 T cells observed in mesenteric lymph nodes (MLN). No age-related changes were observed in the CD4 subset at this site. Furthermore, the percentage of the CD45RC+ cells did not decrease in aged rats, and in the first 9 weeks of life an increase of this subset was observed. PMID- 9397351 TI - Is world activism a reasonable perspective? PMID- 9397352 TI - Work while you learn or learn while you work? PMID- 9397353 TI - An integrated, evidence-based medicine program for FP residents. PMID- 9397354 TI - The invisible faculty. PMID- 9397355 TI - WIN5. PMID- 9397356 TI - The resident's perspective on community rotations. PMID- 9397357 TI - Twenty years of consulting for excellence: the Residency Assistance Program. AB - BACKGROUND AND OBJECTIVES: The Residency Assistance Program (RAP) in family practice was established in 1975 to provide consultative assistance to family practice residency program directors interested in enhancing the quality of their training programs. Since its inception, RAP activities have been monitored and policies approved by a project board, with representation from all the national family medicine/practice organizations. The voluntary, confidential, nonpunitive, collaborative problem-solving process has provided more than 800 RAP consultations in RAP's 20 years of operation. This paper reviews the historical development, current status, and future directions of the program. PMID- 9397358 TI - Competency-based education in family practice. AB - BACKGROUND AND OBJECTIVES: Since their inception, family practice residency programs have been designed on a rotation-based format. It has been assumed that by having residents rotate through a series of educational experiences, they would assimilate the skills necessary to effectively serve as a family physician. An alternative approach is based on the attainment of competency, rather than on the completion of a set of experiences. This method of education is known as competency-based education, mastery learning, or, more recently, outcomes-based assessment. Within family medicine, there is a strong interest in the application of competency-based education to family practice residency training. In response to the growing need to discuss these and other related issues, the Society of Teachers of Family Medicine (STFM) Board created the Task Force on Competency based Education. Its mission is to disseminate this educational theory to STFM's membership. This article reviews the theory of competency-based education, describes development of a competency-based curriculum model, and discusses the academic issues surrounding adaptation of this form of education to family practice residency programs. PMID- 9397359 TI - Resident partnerships: a tool for enhancing ambulatory training. AB - BACKGROUND AND OBJECTIVES: This study examined resident partnerships and their effect on graduates' practice patterns. METHODS: The study authors surveyed graduates from a residency program that used resident partnerships. We also surveyed the graduates' current practice partners, and they served as a comparison group. RESULTS: The graduates' response rate was 86%, and their current practice partners' response rate was 61%. Graduates from a partnership program rated themselves better trained for outpatient medicine and more comfortable communicating with other physicians and working within a patient care team; they were also slightly less likely to practice inpatient medicine. Reported benefits during residency included enhanced availability for continuity clinics, more emotional and intellectual support, and more flexible work schedules. CONCLUSIONS: Graduates valued partnerships during their training and reported being better prepared to work with other physicians in ambulatory settings. PMID- 9397360 TI - Perceived characteristics of successful family practice residency maternity care training programs. AB - BACKGROUND AND OBJECTIVES: This study determined the perceived characteristics of family practice residency training programs that produce a high percentage of graduates who provide maternity care. METHODS: We surveyed a Delphi panel of 28 family practice maternity care experts. RESULTS: Consensus was reached after the third survey. The characteristics of the family medicine faculty and teaching service were rated as most important. Other essential characteristics were an adequate obstetrical training volume; mutual respect between obstetric and family medicine faculty and residents; support for family practice maternity care from obstetricians, administration, and nursing staff; and family physicians being accepted in the community as maternity care providers. CONCLUSIONS: Family practice residency programs that produce a high percentage of graduates who provide maternity care have a unique, family practice maternity care-friendly environment. Residency programs wishing to increase the percentage of their graduates who provide maternity care should ensure that their faculty support family practice maternity care, are competent in maternity care, and model maternity care in their own practices. They should strive to ensure an adequate volume of obstetrical cases for resident education and work toward educating patients and local obstetricians, nursing staff, and hospital administration regarding family practice maternity care. PMID- 9397361 TI - Combined residency training in family practice and other specialties. AB - BACKGROUND AND OBJECTIVES: The University of California, Davis residencies in family practice and psychiatry are entering the third year of a combined training track. Graduates of the combined program will be eligible for board certification in both specialties. The combined program has created opportunities for collaborative training with other specialties, such as obstetrics and medical informatics. Barriers and challenges to collaborative training include issues of curriculum integration, establishment of an identity for residents and future graduates, resident stress, acceptance of combined programs, counting of generalist physicians, hospital privileges, fulfillment of unmet societal needs, and improved collaboration at academic health centers. PMID- 9397362 TI - A curriculum for multicultural education in family medicine. AB - BACKGROUND AND OBJECTIVES: To deliver effective medical care to patients from all cultural backgrounds, family physicians need to be culturally sensitive and culturally competent. Our department implemented and evaluated a 3-year curriculum to increase residents' knowledge, skills, and attitudes in multicultural medicine. Our three curricular goals were to increase self awareness about cultural influences on physicians, increase awareness about cultural influences on patients, and improve multicultural communication in clinical settings. Curricular objectives were arranged into five levels of cultural competence. Content was presented in didactic sessions, clinical settings, and community medicine projects. METHODS AND RESULTS: Residents did self-assessments at the beginning of the second year and at the end of the third year of the curriculum about their achievement and their level of cultural competence. Faculty's evaluations of residents' levels of cultural competence correlated significantly with the residents' final self-evaluations. Residents and faculty rated the overall curriculum as 4.26 on a 5-point scale (with 5 as the highest rating). CONCLUSIONS: Family practice residents' cultural knowledge, cross-cultural communication skills, and level of cultural competence increased significantly after participating in a multicultural curriculum. PMID- 9397363 TI - A hypothetical model of the effect of medical education on specialty choice. AB - BACKGROUND AND OBJECTIVES: Using the Theory of Reasoned Action, we propose a model that diagrams medical school characteristics known or hypothesized to influence the process of specialty choice. The medical school characteristics we consider are administrative support, special programs, primary care funding, number and quality of primary care faculty, faculty influence, primary care residencies, committee representation, primary care environment, required time, and student contact. This model provides explicit hypotheses to be tested in future research on specialty choice. PMID- 9397366 TI - Pediatric nosocomial infections: children are not little adults. PMID- 9397365 TI - Assessing prenatal care in a family practice residency clinic. AB - BACKGROUND AND OBJECTIVES: This study demonstrates how one family practice residency clinic characterized obstetric clinic patients and assessed obstetric care using birth certificate data (demographic characteristics and risk factors) and birth outcome indicators. METHODS: We compared clinical characteristics and birth outcomes for 901 patients who were delivered by family physicians from the family practice residency clinic with a matched and unmatched group of patients who were delivered by other physicians in the county during 1990-1993. RESULTS: The study clinic patients were at higher risk and had lower use of prenatal care. However, the outcomes of the study clinic patients were significantly better (fewer labor and delivery complications, procedures, Cesarean deliveries, abnormal conditions of newborn, low birth weight deliveries, and preterm birth) or no different from the comparison group of non-clinic patients. CONCLUSIONS: The analysis of birth certificate data provided a favorable assessment of prenatal care provided by a family practice residency clinic. This type of analysis permits comparisons of birth outcomes with other local or regional providers, statewide providers, and the year 2000 national objectives established by the National Center for Health Statistics. PMID- 9397364 TI - Factors associated with primary care residents' satisfaction with their training. AB - BACKGROUND AND OBJECTIVES: Satisfaction is known to impact work performance, learning, recruitment, and retention. This study identifies the factors associated with primary care residents' satisfaction with their training. METHODS: We used a cross-sectional survey based on the Price-Mueller model of job satisfaction. The model included 14 job characteristics, four personal characteristics, and four demographic factors. Data were collected in February and March 1996 from residents in three primary care training programs (family practice, pediatrics, and internal medicine) at a large academic medical center. The same standardized, self-administered questionnaires were used in all three departments. RESULTS: Seventy-five percent (n = 119) of the residents returned questionnaires. Five job characteristics were positively associated with resident satisfaction: continuity of care, autonomy, collegiality, work that encourages professional growth, and work group loyalty. Role conflict, a sixth job characteristic, was negatively associated with satisfaction. The personal characteristic of having an optimistic outlook on life was also positively associated with satisfaction. The model explained 66% of the variation in self reported satisfaction. CONCLUSIONS: The satisfaction of the residents was significantly associated with six job characteristics and one personal factor. Interventions based on these job characteristics may increase resident satisfaction and may lead to better patient outcomes, better work performance, greater patient satisfaction, and more success in recruiting top students into a residency. PMID- 9397367 TI - Molecular markers for differentiation of multiresistant Klebsiella pneumoniae isolates in a pediatric hospital. AB - OBJECTIVE: To study the spread of extended-spectrum beta-lactamase-producing, but aminoglycoside-susceptible, Klebsiella pneumoniae strains in our hospital over an 8-month period, by using two genotypic markers. DESIGN: Ribotyping (using two endonucleases) and randomly amplified polymorphic DNA analysis (RAPD; using two different 10-mer primers) were applied to the epidemiological typing of clinical K pneumoniae isolates from stools, ileal fluid, or urine of hospitalized children. SETTING AND PATIENTS: The surgical intensive-care ward (S1: 9 patients, 17 isolates), surgical unit (S2: 2 patients, 2 isolates), and gastroenterology ward (GE: 1 patient, 1 isolate) of the Robert Debre Hospital of Paris, France. RESULTS: Ribotyping of the 20 clinical isolates, the type strain of the species, and two epidemiologically unrelated isolates with EcoRI and HindIII revealed 6 and 5 different patterns, respectively. Six ribotypes were identified by using these two enzymes. RAPD generated 6 distinct patterns, in complete agreement with ribotyping. Our genotypic results showed that 11 patients from wards S1, S2, and GE harbored genotypically related strains, suggesting nosocomial transmission and cross-colonization between and within the three wards. CONCLUSIONS: Ribotyping and RAPD appear to be reliable methods for distinguishing K pneumoniae strains. The spread of one strain of K pneumoniae in different units of our hospital was demonstrated by both methods. However, RAPD has the advantage of simplicity and rapidity conferred by polymerase chain reaction. PMID- 9397368 TI - Nosocomial pneumonia on general medical and surgical wards in a tertiary-care hospital. AB - OBJECTIVE: To describe the demographic, clinical, and microbiologic characteristics of patients who develop nosocomial pneumonia on general medical and surgical wards of a tertiary-care hospital. DESIGN: A 1-year, prospective, descriptive study. SETTING: A 1,100-bed, tertiary-care, urban hospital. POPULATION: Patients experiencing nosocomial pneumonia were identified through surveillance on general medical and surgical wards, using a standard case definition. RESULTS: 92 pneumonias in 85 patients on general wards were identified. The mean age of patients was 63 +/- 17 years, 55 patients (65%) were male, and 75 cases of pneumonia (81%) were acquired on surgical wards. Bacteremia was identified in 8 (13%) of 62 episodes, and 48 (52%) grew potential pathogens from respiratory specimens. Twenty-six patients (28%) required transfer to the intensive-care unit (ICU), and 20 (22%) received mechanical ventilation. By multivariate analysis, patients with a thoracic surgical procedure or with Staphylococcus aureus isolated from respiratory secretions were more likely to require ICU admission. The overall mortality rate was 20% (17/85), with a directly associated mortality of 14% (12/85). Patients who died were older, more frequently resided on a medical ward, and had a greater mean number of comorbidities. These patients often were treated nonaggressively and were not considered candidates for ICU admission due to advanced age and poor underlying clinical status. CONCLUSIONS: Although the morbidity of nosocomial pneumonia in this population was high, as evidenced by high rates of transfer to ICU, the directly associated mortality was relatively low. Those requiring ICU admission require further study to identify preventive measures that could decrease the morbidity in this group. Interventions to prevent pneumonia or to improve prognosis may not be feasible for the majority of these patients who die from nosocomial pneumonia. PMID- 9397369 TI - Risk factors associated with permanent access-site infections in chronic hemodialysis patients. AB - OBJECTIVE: To determine the epidemiological risk factors associated with permanent access-site (PAS) infection in a population of chronic hemodialysis patients. DESIGN: Retrospective cohort analysis. SETTING: Hemodialysis unit of a 400-bed Department of Veterans' Affairs hospital. RESULTS: A cohort of 94 males (1,316 patient months) was studied. Fifty-one PAS infections in 31 patients were observed. Six patients had two PAS infections, four patients had three infections, and two patients had four infections. Twenty-nine of the 31 patients with PAS infections were bacteremic at least once. Univariate analysis identified seven factors significantly associated with PAS infection in this population: location of PAS other than forearm, type of vascular access (polytetrafluoroethylene [PTFE] versus endogenous arteriovenous [AV] fistula), limited ambulatory status, residence in a nursing home, bacterial infection at a distant site, number of access-site revisions, and number of hospitalizations. In a logistic regression analysis, only graft type and number of PTFE graft revisions were associated independently with PAS infection. The odds ratio for PAS infection in PTFE grafts compared to endogenous AV fistula was 7.8; the odds ratio for PAS infection with each PTFE graft revision was 1.5. CONCLUSIONS: PAS infections were associated independently with the type of graft and the number of PTFE graft surgical revisions. PMID- 9397370 TI - The epidemiology of fecal carriage of vancomycin-resistant enterococci. AB - An outbreak of vancomycin-resistant enterococci (VRE) began at the University of Massachusetts Medical Center in May 1993. As of September 1995, we had a total of 253 patients infected or colonized with VRE, with consequent increasing demand for private rooms. We analyzed results of surveillance cultures for VRE of 49 patients known to be colonized or infected with VRE. Of these, 34 (70%) were classified as persistent carriers, defined as patients with at least three consecutively positive cultures from any site taken over at least a 2-week period. The length of carriage varied from 19 to 303 days (median, 41 days); 11 patients were converters, defined as patients with three consecutive negative cultures from all previously colonized sites taken over a 3-week period. These patients were free of VRE for 39 to 421 days (median, 142 days). Four were recolonizers after they were documented to be clear of VRE for 33 to 106 days. VRE carriage tends to be prolonged, and hospitalization of patients with VRE will require continued isolation and contact precautions for control of transmission. PMID- 9397371 TI - Living with methicillin-resistant Staphylococcus aureus: a 7-year experience with endemic MRSA in a university hospital. AB - A review of our infection control records revealed 3,159 new isolations of methicillin-resistant Staphylococcus aureus (MRSA) from 1988 to 1994. Prior to this period, our approach to MRSA had changed from eradication to containment measures. We found a decline in MRSA rates from 11.4 to 5.2 first isolations per 1,000 deaths and discharges over the study period. PMID- 9397372 TI - Prescribing pattern of vancomycin in a community teaching hospital with low prevalence of vancomycin-resistant enterococci. AB - A 1-month prospective survey of all inpatients given vancomycin was performed in a community teaching hospital with a low prevalence of vancomycin-resistant enterococci. Only 20 of the 97 vancomycin orders written from August 1 to September 1, 1996, were consistent with Hospital Infection Control Practices Advisory Committee (HICPAC) guidelines. Surgical prophylaxis accounted for 37 of the 77 orders inconsistent with HICPAC guidelines. PMID- 9397373 TI - Ribotyping and random amplification of polymorphic DNA for nosocomial Enterobacter cloacae isolates in a pediatric intensive-care unit. AB - Between 1987 and 1989, two sequential outbreaks of nosocomial infection caused by Enterobacter cloacae occur-red in the pediatric intensive-care unit of a tertiary care teaching hospital. Seventeen strains retrieved from the outbreaks and two control strains identified in other wards were typed by ribotyping and random amplification of polymorphic DNA (RAPD). The results indicated that the genomic pattern of strains identified between the first and second outbreaks was different. We conclude that both ribotyping and RAPD are highly discriminatory and reproducible methods for typing E cloacae. RAPD seems to be more efficacious and cost-effective. PMID- 9397374 TI - Epidemiological study of hospital-acquired infection with vancomycin-resistant Enterococcus faecium: possible transmission by an electronic ear-probe thermometer. AB - Clonal spread of vancomycin-resistant Enterococcus faecium among seven patients on one ward of a community teaching hospital was identified by contour-clamped homogeneous electric-field gel electrophoresis. Environmental cultures isolated the same strain from the handle of a shared electronic ear-probe thermometer. Cross-contamination of the clonal strain between two geographically separate units on this ward, sharing equipment but not personnel, suggests the possibility of an environmental source. PMID- 9397375 TI - A cluster of serious Escherichia coli infections in a neonatal intensive-care unit. AB - A cluster of serious Escherichia coli infections was identified among patients in a neonatal intensive-care unit. Infection control staff identified the outbreak because they realized that E coli rarely caused infections in this unit. Pulsed field gel electrophoresis confirmed that one strain of E coli was transmitted among patients. PMID- 9397376 TI - Cumulative yield from patient surveillance cultures for methicillin-resistant Staphylococcus aureus during a hospital outbreak. AB - The cumulative yield from cultures of separate sites was determined during the investigation of a methicillin-resistant Staphylococcus aureus (MRSA) outbreak. Surveillance cultures were submitted from clinical sites, nose, groin, and axilla of 421 patients on two different occasions. MRSA was recovered most often from various clinical sites, including lower respiratory tract, surgical wounds, urinary tract, and decubitus ulcers (total, 13 patients). Four additional patients were identified as positive from the first nasal swab, one patient from the second nasal swab, and two others from swabs of the groin. The submission of axillary swabs or a second groin swab did not identify additional MRSA-colonized patients and resulted in additional costs of $4,525. PMID- 9397377 TI - Rate of seasonal spread of respiratory syncytial virus in a pediatric hospital. AB - The rate of nosocomial respiratory syncytial virus (RSV) infection was measured in a large pediatric hospital using an incidence density method. The at-risk days for nosocomial RSV were summed during a defined winter period in which there were 54 admissions with community-acquired RSV infection giving a rate of 2.9 cases per 1,000 at-risk days (95% confidence interval, 0.3-5.4 per 1,000). PMID- 9397378 TI - Evaluation of vancomycin use in a pediatric teaching hospital based on CDC criteria. AB - This study was performed to determine whether vancomycin use at our pediatric hospital was consistent with modified Centers for Disease Control and Prevention guidelines. Vancomycin use was inappropriate in 54% of patients. Inappropriate use briefly decreased by 14% after educational efforts. Further education regarding vancomycin use was deemed necessary and is continuing. PMID- 9397379 TI - Hospital-acquired pneumonia: perspectives for the healthcare epidemiologist. AB - Nosocomial pneumonia is defined as an infection of lung parenchyma that was neither present nor incubating at the time of the patient's admission to the hospital. In the United States, hospital-acquired pneumonia is the second most common nosocomial infection and accounts for the most deaths from nosocomial infection. We describe how infection control personnel can use targeted surveillance to identify clusters of cases and to prevent pneumonia. We also discuss common pathogens that cause nosocomial pneumonia; ventilator-associated pneumonia; and strategies for prevention of hospital-acquired pneumonia. PMID- 9397380 TI - Outbreak of highly contagious tuberculosis. PMID- 9397381 TI - Disease transmitted through food supply. PMID- 9397382 TI - Benefits of hormone replacement therapy--overview and update. AB - Postmenopausal estrogen deficiency may result in a wide variety of physiologic disorders, including vasomotor symptoms, urogenital atrophy, an increase in the risk of coronary heart disease, osteoporotic fractures, and Alzheimer's disease. The growing body of evidence, including much that is newly published, demonstrating that hormone replacement therapy (HRT) can largely prevent or mitigate these sequelae, will be reviewed in this paper. The efficacy of HRT in alleviating vasomotor and urogenital discomfort, the most common symptoms of postmenopausal estrogen deficiency, is well established. Evidence from over 30 epidemiologic studies indicates that estrogen reduces the risk of coronary heart disease (CHD) by 50%. The risk of major CHD has been found to be markedly reduced in women who receive combined estrogen/progestogen therapy compared to nonusers (or estrogen-alone users). Estrogen is recommended as the modality of choice to prevent bone loss: data supporting a positive effect of estrogen on the risk of wrist and vertebral fracture are quite favorable. Similarly, outcomes of recent investigations have demonstrated a positive impact of HRT on both psychological function and the risk of osteoarthritis. In addition, HRT substantially reduces the risk of colon cancer. Moreover, the potential for HRT to delay the progression or reduce the risk for developing Alzheimer's disease is a new area of research that shows promise. Improvements in quality-of-life assessments have also been reported in conjunction with the relief of menopausal symptoms by HRT. Clinicians should be aware of the large amount of new evidence that strengthens the case for wider use of HRT. Based on these new data, physicians may conclude that HRT would benefit the majority of their postmenopausal patients and thus encourage HRT use in the absence of known risk factors. PMID- 9397383 TI - Evaluation and management of the hormone replacement therapy (HRT) candidate. AB - As knowledge about menopause and hormone replacement therapy (HRT) has increased, it has become evident that a considerably higher percentage of postmenopausal women than the 20% to 30% currently treated could receive HRT. It is equally clear, however, that HRT is not appropriate for every woman and that "one size fits all" management of menopausal women is not always suitable. This paper, therefore, reviews published prescribing and management guidelines for instituting and maintaining HRT and summarizes current information concerning factors to be considered before recommending HRT. When choosing candidates, special attention should be placed on individual patient factors. A thorough history is required to determine the presence of contraindications and the likelihood of potential benefits and risk factors. The hormone replacement regimen, hormone preparations, and dosage forms that best meet the specific needs of the individual woman should be offered. Before undertaking long-term therapy, the candidate should be informed of the established and likely benefits and the relative risks of hormone therapy, and that the magnitude of some risks has not yet been definitively determined. The final decision to use therapy should be made by the patient, guided by her physician, and based on her current symptoms and her relative likelihood of developing coronary artery disease, osteoporotic fractures, and cancer. The ancillary benefits, the common side effects of each regimen, the bleeding patterns to expect, and the type and frequency of clinical monitoring that will be necessary during therapy should also be considered. PMID- 9397384 TI - Cross-national study of women's use of hormone replacement therapy (HRT) in Europe. AB - The benefits of hormone replacement therapy (HRT) to perimenopausal and postmenopausal women are well established. Women from France, Germany, Spain, and the United Kingdom were interviewed to determine: (1) knowledge and views of menopause and HRT; (2) history of HRT among current and lapsed users; and (3) reasons of non-users for never having taken HRT. In 1996, nearly 1,500 women aged 40 to 65 answered a short series of questions concerning menopause and HRT as part of a consumer omnibus survey; others (n = 929) participated in a focused survey of attitudes toward and use of HRT. Only one-third of perimenopausal and 13% of postmenopausal women currently were taking HRT. About 25% of postmenopausal women reported having taken HRT at some time. The proportion of perimenopausal women using HRT varied by country, and ranged from 18% in Spain to 55% in France. Importantly, about half (range across countries, 38% to 61%) of the women interviewed had not discussed menopause or its symptoms with their doctors. While levels of HRT knowledge varied by country, two-thirds of respondents overall believed they needed more information about HRT. Decisions about beginning HRT and choosing a formulation were viewed by most women as matters of personal choice, to be made with advice from a physician. In summary, despite the benefits of HRT and available choices among drug delivery options, a fairly small proportion of European women use it, largely because most remain poorly informed about the therapy. Increased physician-patient communication and public education programs are needed to provide women with the information they need to make judicious decisions concerning HRT. PMID- 9397385 TI - Comparison of continuous and sequential transdermal progestogen with sequential oral progestogen in postmenopausal women using continuous transdermal estrogen: vasomotor symptoms, bleeding patterns, and serum lipids. AB - OBJECTIVE: The efficacy, bleeding patterns, and safety of continuous transdermal and sequential transdermal progestogen therapy were compared with those of oral progestogen therapy in postmenopausal women receiving transdermal estrogen. METHODS: In an open-label, 1-year (13 treatment periods, 28 days each), randomized study, 774 postmenopausal women were assigned to receive 50 micrograms/day of continuous trans dermal estradiol with either continuous or sequential transdermal norethisterone acetate (NETA) in daily doses of 170 or 350 micrograms in a single transdermal patch or sequential oral progestogen (1 mg norethisterone [NET] or 20 mg dydrogesterone/day). RESULTS: The average number of hot flushes/day decreased from prestudy by over 90% (P < .001), and this reduction was unaffected by different progestogen regimens. With sequential progestogen, bleeding incidence and the number of bleeding days did not change over the course of the study but were lower in the low-dose transdermal progestogen group. With continuous progestogen, the incidence of bleeding decreased in both the low- and high-dose groups, from 35% and 45% in treatment period 1 to 25% and 15%, respectively, at the end of treatment. Adverse event incidence was similar in all groups, with 23% to 36% of subjects reporting events possibly or probably related to HRT (excluding vaginal bleeding). Two cases of simple hyperplasia were reported (one in each low-dose progestogen group). Beneficial effects on coronary heart disease risk factors, such as reductions in total cholesterol and low-density lipoprotein cholesterol and increases in high density lipoprotein-2 cholesterol levels, were measured in all treatment groups. Lipoprotein (a) was reduced in all but the oral progestogen group. CONCLUSIONS: Continuous and sequential transdermal estrogen/progestogen treatments with estradiol/NETA appear to be effective and safe alternatives to continuous transdermal estrogen and oral sequential progestogen for the treatment of menopausal symptoms. Continuous transdermal therapy with estradiol/NETA may be more acceptable for a majority of patients, i.e., those who wish to avoid monthly bleeds, whereas the sequential regimen may be preferable when the clinician and/or patient believes monthly bleeding to be appropriate. PMID- 9397386 TI - Transdermal sequential and continuous hormone replacement regimens with estradiol and norethisterone acetate in postmenopausal women: effects on the endometrium. AB - OBJECTIVE: To evaluate, in postmenopausal women, the endometrial safety and histologic effects of two doses of transdermal norethisterone acetate (NETA) administered in sequential and continuous treatment regimens added to continuous transdermal estradiol, against a reference regimen consisting of sequential oral progestogen and continuous transdermal estradiol. METHODS: A total of 774 postmenopausal women were enrolled in the open-label study of 13 treatment cycles of 28 days each and randomly assigned evenly to regimens consisting of transdermal estradiol, 50 micrograms/day and NETA, given sequentially (last 14 days of each treatment cycle) or continuously at two doses (170 and 350 micrograms/day). Estradiol and NETA were delivered from a transdermal system containing both hormones. The reference group received estradiol, 50 micrograms/day transdermally and either 1 mg/day NET or 20 mg/day dydrogesterone orally during the last 14 days of each treatment cycle. Endometrial biopsies were taken pre-study and at the end of the treatment, if treatment had lasted at least 3 months. Safety was to be assessed in terms of the incidence of hyperplasia. Endometrial biopsies were assigned to one of the following histological classes: proliferative (predominant estrogen effect), suppressed proliferation (slightly, moderately, strongly progestogenic effect), progestational atrophy (predominant progestogenic effect), hyperplastic, cancerous, or other. RESULTS: No case of hyperplasia was recorded in any of the treatment groups, each with > 150 subjects enrolled, after one year of treatment. One case of serous carcinoma was found in the LP-C group. Progestational atrophy was seen frequently in women receiving continuous transdermal HRT (84%, high-dose NETA, 66%, low-dose NETA); it was much rarer with the sequential regimens (i.e., between 32% and 38%). The proportion of estrogen-dominated endometria was low, 0.9% and 2.6% with the high- and low-dose NETA continuous regimens, respectively, 6.2% and 12.5% with the high- and low dose NETA sequential regimens, respectively; and, 4.5% in the oral progestogen group. CONCLUSION: Transdermal administration of either dose of NETA, whether given sequentially or continuously, prevents the emergence of hyperplasia expected with unopposed estradiol. Since differences in outcomes of endometrial histology between the two NETA doses were minor for both continuous and sequential regimens, use of the lower NETA dose is considered sufficient for a safe transdermal combination hormone replacement therapy. PMID- 9397387 TI - Short-term and working memory differences in language/learning disabled and normal adults. AB - Fifteen adults who reported a childhood history of speech-language and/or learning disability (L/LD) were tested on two verbal memory tasks. Their performance on sentence repetition and reading span measures was compared with that of a matched control group who reported no childhood history of L/LD. Results indicated statistically significant group performance differences on both short-term and working memory tasks. This suggests that verbal memory difficulties may be a longterm component of L/LD. PMID- 9397388 TI - Effect of sign task on speech timing in simultaneous communication. AB - The purpose of this investigation was to study the effect of the signing task on temporal features of speech during simultaneous communication (SC). The effects of three independent variables: (a) communication mode (speech only vs. SC); (b) sign task demand (base vs. elaborated signs); and (c) type of sign movement (kinetic vs. morphokinetic) were studied on five dependent variables: (a) word duration; (b) sentence duration; (c) diphthong duration; (d) interword interval before signed experimental word (IWIB); and (e) interword interval after signed experimental word (IWIA). Audio recordings were made of 12 normal hearing, experienced sign language users speaking experimental words that varied in sign task demand and movement under SC and speech only (SO) conditions. Results indicated longer sentence durations for SC than SO and longer anticipatory durations of IWIB and diphthong before signed words, especially those using signs with greater task demand or with movements including hand shape change. IWIA only lengthened for SC vs. SO with no further effect of sign task demand or movement. These results indicate finite effects of sign task demand and movement on pause and segment durations before the sign but not as strong an effect as has been reported for increased finger spelling task length. PMID- 9397389 TI - Maintaining acceptably low referral rates in TEOAE-based newborn hearing screening programs. AB - This article describes factors that can affect the refer rate for otoacoustic emission (OAE) based newborn hearing screening, including the population of infants being screened, the adequacy of probe fit, software options used, external ear conditions, screener training, and baby handling. The effect of the infant's age on screening outcomes is also discussed using results of screening for 1328 regular nursery newborns, ranging in age from 6 to 60 hours, who were screened with transient evoked otoaoustic emissions (TEOAE) prior to hospital discharge. The youngest infants (6-9 hours old) were as likely to pass (90% pass rate) as the infants who were 24-27 hours old (94% pass rate). The results of this study are consistent with reports from many TEOAE-based screening programs that have demonstrated that acceptably low refer rates (mean = 6.9%) can be obtained when appropriate screening procedures are followed. PMID- 9397390 TI - Cost analysis of TEOAE-based universal newborn hearing screening. AB - Although more and more hospitals are implementing universal newborn hearing screening programs, there is still very little information available about the costs of newborn hearing screening programs. The few articles which have been published evaluate technologies or protocols which are no longer used, are incomplete, or are based on hypothetical estimates of the costs and time necessary to do screening. After briefly reviewing the extant literature, this article describes a cost analysis of a TEOAE-based universal newborn hearing screening program. Reasons why the cost per baby ($7.42) is lower than in previous reports are explained, and the benefits of having accurate cost analysis data are summarized. PMID- 9397391 TI - Production and perception of final consonant voicing in speech during simultaneous communication. AB - Simultaneous communication combines both spoken and manual modes to produce each word of an utterance. This study investigated the potential influence of alterations in the temporal structure of speech produced during simultaneous communication on the perception of final consonant voicing. Experienced signers recorded words that differed only in the voicing characteristic of the final consonant under two conditions: (a) speech alone and (b) simultaneous communication. The words were digitally edited to remove the final consonant and played to 20 listeners who, in a forced-choice paradigm, circled the word they thought they heard. Results indicated that accurate perception of final consonant voicing was not impaired by changes in the temporal structure of speech that accompany simultaneous communication. PMID- 9397392 TI - Objective and subjective time courses of recovery from motion sickness assessed by repeated motion challenges. AB - The aim of this study was to determine whether the time course of recovery of tolerance, as assessed objectively by rechallenge with motion, paralleled the subjective recovery from motion sickness. Subjects (n = 20) were exposed to 5 pairs of nauseogenic motion challenges in which the time interval between the end of the first and the start of the second of each pair ranged from 15 min to 2 h. The cross-coupled motion challenge had an incrementing profile of rotational velocity from 4 degrees to 92 degrees.s-1 in steps of 4 degrees.s-1 every 30 s, with 8 head movements per 30 s, of approximately 45 degrees, and was continued to the point of moderate nausea. Objective loss of tolerance decreased from 15 min to 60 min after the first challenge, but increased again at 2 h. By contrast, most individuals reported subjective recovery by 15 min to 30 min. It was concluded that there is an underlying effect of motion sickness that sensitizes the response to subsequent motion for a period of at least 2 h. This underlying objective effect can occur in the absence of subjective symptoms, has a slower time course than the subjective recovery from symptoms, and appears to be non monotonic. PMID- 9397393 TI - Orientation illusions in spaceflight. AB - Investigations of spontaneous illusory reactions were carried out during space flights of various durations by ANKETA questionnaires (104 cosmonauts). From a total of 104 cosmonauts, 24, in addition, used a dictaphone to record a verbal description of the illusions and their sensations on tape. Analysis of data generated by ANKETA and the tapes thus recorded have shown that during adaptation to weightlessness, 98% of cosmonauts have noted the occurrence of various illusions of orientation (coordinate and kinetic): illusions pertaining to their position or illusions of self- and surround-motion. The development of spatial orientation illusions during and after flight is not limited to certain individuals, but is a general response of the sensory system to microgravity. These responses differ to some extent among individuals in severity, nature, time and duration of occurrence, and the dynamics of the process. Perceptual disorders may occur even if the cosmonaut feels well and experiences no anomalous autonomic reactions. The nature of spatial illusions was determined by the role and relative contribution of various types of sensory input to spatial orientation. After completion of the initial stage of adaptation to weightlessness, the perceptual disorders disappear. However, spontaneous illusory reactions were often observed after 50 days of exposure to weightlessness. The adaptation process during long-term spaceflight had an undulating course, in which adaptation and de-adaptation alternated. A classification of weightlessness illusions is proposed. PMID- 9397394 TI - The velocity storage mechanism of the optokinetic nystagmus under apparent stimulus movements in squirrel monkeys. AB - Experiments in two awake untrained squirrel monkeys were performed to study the velocity storage mechanism during fast rise of OKN slow phase velocity. This was done by testing the monkey's capability to perform OKN in response to a stationary-appearing stroboscopically illuminated stripe pattern of a horizontally rotating drum. Nystagmus was initially elicited during constant illumination lasting between 0.6 and 25 s. The periodicity of the stripe pattern was 2.37 degrees. When after the constant light the flash illumination was switched on again, two types of behavior could occur, depending on the length of the constant light interval (CLI): 1) when the CLI was shorter than a threshold value of 6.2 seconds, the OKN ceased under the flash stimulation. Then a "post OKN" occurred that increased with the length of the CLIs, indicating that the intermittently illuminated pattern did not provoke fixation suppression of OKN aftereffects. 2) when the CLI was above threshold, the OKN continued under the flash light; it will be called "apparent movement OKN." The threshold CLI between the type 1 and the type 2 response did not depend on drum velocities between 21.5 degrees/s and 71.3 degrees/s. The average gain of the apparent movement OKN was 0.83 +/- 0.04; gain and stability of slow phase eye movement velocity did not deviate systematically from the usually elicited OKN. OKAN after apparent movement OKN did not deviate from OKAN after constantly illuminated moving patterns. In response to the OKN initiation by a constantly illuminated pattern up to pattern velocities of 100 degrees/s, the OKN steady state gain was reached within the first 2 or 3 nystagmus beats. We ascribe the increase of the post-OKN with CLI and the existence of a threshold constant light interval to activity accumulation in the common velocity-to-position integrator (velocity storage) of the brain stem. Loading of the velocity storage takes place after the OKN gain has already reached the steady-state value. Apparent movement OKN could also be elicited in guinea pigs that lack an effective smooth pursuit system. We suggest that apparent movement OKN is produced by mechanisms located in the brain stem. PMID- 9397395 TI - Microgravity vestibular investigations: perception of self-orientation and self motion. AB - Four astronauts experienced passive whole-body rotation in a number of test sessions during a 7-day orbital mission. Pitch (Y-axis) and roll (X-axis) rotation required subject orientations on the rotator in which the otolith system was at radius of 0.5 m. Thus subjects experienced a constant -0.22 Gz stimulus to the otoliths during the 60 s constant-velocity segments of "pitch" and "roll" ramp profiles. The Gz stimulus, a radius-dependent vector ranging from -0.22 Gz at the otoliths to +0.36 Gz at the feet, generated sensory information that was not interpreted as inversion in any of the 16 tests carried out in flight (12 in pitch and 4 in roll orientation). None of the subjects was rotated with head off center during the first 33 h of the mission. In the state of orbital adaptation of these subjects, a -0.22 Gz otolith stimulus did not provide a vertical reference in the presence of a gradient of +Gz stimuli to the trunk and legs. PMID- 9397396 TI - Vestibular bibliography. PMID- 9397397 TI - The effect of dietary docosahexaenoic acid on platelet function, platelet fatty acid composition, and blood coagulation in humans. AB - The effect of dietary docosahexaenoic acid (DHA) in the absence of eicosapentaenoic acid (EPA) has been studied infrequently in humans under controlled conditions. This 120-d study followed healthy, adult male volunteers who lived in the metabolic research unit (MRU) of the Western Human Nutrition Research Center for the entire study. The basal (low-DHA) diet consisted of natural foods (30 en% fat, 15 en% protein, and 55 en% carbohydrate), containing < 50 mg/d of DHA, and met the recommended daily intake for all essential nutrients. The high-DHA (intervention) diet was similar except that 6 g/d of DHA in the form of a triglyceride containing 40% DHA replaced an equal amount of safflower oil in the basal diet. The subjects (ages 20 to 39) were within -10 to +20% of ideal body weight, nonsmoking, and not allowed alcohol in the MRU. Their exercise level was constant, and their body weights were maintained within 2% of entry level. They were initially fed the low-DHA diet for 30 d. On day 31, six subjects (intervention, group A) were placed on the high-DHA diet; the other four subjects (controls, group B) remained on the low-DHA diet. Platelet aggregation in platelet-rich plasma was determined using ADP, collagen, and arachidonic acid. No statistical differences could be detected between the amount of agonist required to produce 50% aggregation of platelet-rich plasma before and after the subjects consumed the high-DHA diet. The prothrombin time, activated partial thromboplastin time, and the antithrombin-III levels in the subjects were determined, and, again, there were no statistically significant differences in these three parameters when their values were compared before and after the subjects consumed the high-DHA diet. In addition, the in vivo bleeding times did not show any significant difference before and after the subjects consumed the high-DHA diet (9.4 +/- 3.1 min before and 8.0 +/- 3.4 min after). Platelets from the volunteers exhibited more than a threefold increase in their DHA content from 1.54 +/- 0.16 to 5.48 +/- 1.21 (wt%) during the DHA feeding period. The EPA content of the subjects' platelets increased from 0.34 +/- 0.12 to 2.67 +/- 0.91 (wt%) during the high-DHA diet despite the absence of EPA in the subjects' diets. The results from this study on blood clotting parameters and in vitro platelet aggregation suggest that adding 6 g/d of dietary DHA for 90 d to a typical Western diet containing less than 50 mg/d of DHA produces no observable physiological changes in blood coagulation, platelet function, or thrombotic tendencies in healthy, adult males. PMID- 9397399 TI - Pancreatic bile salt-dependent lipase activity in serum of normolipidemic patients. AB - Bile salt-dependent lipase (BSDL, E.C. 3.1.1.-) is a digestive enzyme secreted by the pancreatic acinar cell. Once in the duodenum, the enzyme, upon activation by primary bile salts, hydrolyzes dietary lipid esters such as cholesteryl esters and lipid-soluble vitamin esters. This enzyme is partially transferred from the duodenum or pancreas to the circulation where it has been postulated to exert a systemic action on atheroma-generating oxidized-low density lipoprotein (LDL). In the present study, sera from 40 healthy normolipidemic volunteers were used to investigate the possible linkage between circulating BSDL, lipids, and lipoproteins. We showed, firstly, that pancreatic-like BSDL activity can be detected in these serums. Secondly, BSDL activity increased significantly with the level of LDL-cholesterol and was also positively linked to the serum concentration of Apo B100 and Apo A-I. Thirdly, we also established that BSDL was associated with LDL, in part by a specific interaction with Apo B100, while no interaction was found with Apo A-I. No linkage with other recorded parameters (triglycerides, phospholipids, and high density lipoprotein-cholesterol) was detected. Because an increase in LDL-cholesterol represents an important risk factor for atheroma, the concomitant increase in BSDL, which can metabolize atherogenic LDL, suggests for the first time that this circulating enzyme may exert a positive effect against atherosclerosis. PMID- 9397398 TI - The effect of dietary docosahexaenoic acid on plasma lipoproteins and tissue fatty acid composition in humans. AB - Normal, healthy male volunteers (n = 6) were fed diets [high docosahexaenoic acid DHA] containing 6 g/d of DHA for 90 d. The stabilization (low-DHA) diet contained less than 50 mg/d of DHA. A control group (n = 4) remained on the low-DHA diet for the duration of the study (120 d). Blood samples were drawn on study days 30 (end of the stabilization period), 75 (midpoint of the intervention period), and 120 (end of the intervention period). Adipose tissue (AT) samples were taken on days 30 and 120. The plasma cholesterol (C), low density lipoprotein (LDL)-C and apolipoproteins (apo) [Al, B, and lipoprotein (a)] were unchanged after 90 d, but the triglycerides (TAG) were reduced from a mean value of 76.67 +/- 24.32 to 63.83 +/- 16.99 mg/dL (n = 6, P < 0.007 using a paired t-test) and the high density lipoprotein (HDL)-C increased from 34.83 +/- 4.38 mg/dL to 37.83 +/- 3.32 mg/dL (n = 6, P < 0.017 using a paired t-test). The control group showed no significant reduction in plasma TAG levels. Apo-E, however, showed a marked increase in the volunteers' plasma after 90 d on the high-DHA diet, from 7.06 +/- 4.47 mg/dL on study day 30 to 12.01 +/- 4.96 mg/dL on study day 120 (P < 0.002 using a paired t-test). The control subjects showed no significant change in the apo-E in their plasma (8.46 +/- 2.90 on day 30 vs. 8.59 +/- 2.97 on day 120). The weight percentage of plasma DHA rose from 1.83 +/- 0.22 to 8.12 +/- 0.76 after 90 d on the high-DHA diet. Although these volunteers were eating a diet free of eicosapentaenoic acid (EPA), plasma EPA levels rose from 0.38 +/- 0.05 to 3.39 +/ 0.52 (wt%) after consuming the high-DHA diet. The fatty acid composition of plasma lipid fractions--cholesterol esters, TAG, and phospholipid--showed marked similarity in the enrichment of DHA, about 10%, after the subjects consumed the high-DHA diet. The DHA content of these plasma lipid fractions varied from less than 1% (TAG) to 3.5% (phospholipids) at baseline, study day 30. EPA also increased in all plasma lipid fractions after the subjects consumed the high-DHA diet. There were no changes in the plasma DHA or EPA levels in the control group. Consumption of DHA also caused an increase in AT levels of DHA, from 0.10 +/- 0.02 to 0.31 +/- 0.07 (wt%) (n = 6, P < 0.001 using a paired t-test), but the amount of EPA in their AT did not change. Thus, dietary DHA will lower plasma TAG without EPA, and DHA is retroconverted to EPA in significant amounts. Dietary DHA appears to enhance apo-E synthesis in the liver. It appears that DHA can be a safe and perhaps beneficial supplement to human diets. PMID- 9397400 TI - Epicoprostanol found in adipocere from five human autopsies. AB - Adipocere formation is well known as a later postmortem change. We collected adipocere from five male victims which had been submerged under the sea or fresh water for 1 mon to 4 yr. Fresh subcutaneous fat of a male victim who died from a cerebral contusion was used as the control. The samples were homogenized, and the lipids were extracted with chloroform and methanol followed by injection into a gas chromatograph and a gas chromatograph-mass spectrometer. We detected hydroxy fatty acids (10-hydroxyoctadecanoic acid and 10-hydroxyhexadecanoic acid) as well as 10-ketooctadecanoic acid in adipocere, but not in the control. In addition, we found for the first time a cholesterol-related peak with a molecular ion of 388 in adipocere and identified it as epicoprostanol, suggesting not only oxidation but also reduction had occurred during the formation of adipocere. In addition, we showed the time-course of epicoprostanol accumulation. The relationship between the time of adipocere formation and the characteristic lipid composition is discussed. PMID- 9397401 TI - Prevention of ischemia-induced cardiac sudden death by n-3 polyunsaturated fatty acids in dogs. AB - The objective of this study was to obtain functional information associated with the prevention by n-3 polyunsaturated fatty acids (PUFA) of ischemia-induced fatal cardiac ventricular arrhythmias in the intact, conscious, exercising dog. Thirteen dogs susceptible to ischemia-induced ventricular fibrillation were prepared surgically by ligation of their anterior descending left coronary artery and placement of an inflatable cuff around their left circumflex artery. After 4 wk of recovery, exercise-plus-ischemia tests were performed without and then with an intravenous infusion of an emulsion of free n-3 PUFA just prior to occluding the left circumflex artery while the animals were running on a treadmill. One week later the exercise-plus-ischemia test was repeated but with a control infusion replacing the emulsion of n-3 PUFA. The infusion of the free n-3 PUFA in quantities of 1.0 to 10 g prevented ventricular fibrillation in 10 of the 13 dogs tested (P < 0.005), apparently without esterification of the PUFA into membrane phospholipids. The antiarrhythmic effect of the n-3 PUFA was associated with slowing of the heart rate, shortening of the QT-interval (electrical action potential duration), reduction of left ventricular systolic pressure, and prolongation of the electrocardiographic atrial-ventricular conduction time (P-R interval). These effects are comparable with those we have reported in studies with cultured neonatal rat cardiac myocytes. PMID- 9397402 TI - Biliary excretion of dolichols and beta-hexosaminidase--effect of ethanol and glucagon. AB - Alcohol has been reported to increase the urinary excretion of dolichols, and urinary dolichols are suggested to be derived from the lysosomes of the renal cells. In the present study we examined the effects of alcohol and glucagon on the biliary excretion of dolichols in rats. Chronic ethanol treatment decreased both biliary dolichol and beta-hexosaminidase excretion. The absolute amount of dolichol excreted into the bile correlated highly significantly with the absolute amount of biliary beta-hexosaminidase. Our results indicate that biliary dolichols are--at least in part--derived from hepatic lysosomes. Decreased biliary dolichol output during chronic alcohol administration suggests that urinary and biliary dolichol excretions are regulated independently of each other. PMID- 9397403 TI - Effect of curcumin and capsaicin on arachidonic acid metabolism and lysosomal enzyme secretion by rat peritoneal macrophages. AB - The inflammatory mediators secreted by macrophages play an important role in autoimmune diseases. Spice components, such as curcumin from turmeric and capsaicin from red pepper, are shown to exhibit antiinflammatory properties. The influence of these spice components on arachidonic acid metabolism and secretion of lysosomal enzymes by macrophages was investigated. Rat peritoneal macrophages preincubated with 10 microM curcumin or capsaicin for 1 h inhibited the incorporation of arachidonic acid into membrane lipids by 82 and 76%: prostaglandin E2 by 45 and 48%; leukotriene B4 by 61 and 46%, and leukotriene C4 by 34 and 48%, respectively, but did not affect the release of arachidonic acid from macrophages stimulated by phorbol myristate acetate. However, the secretion of 6-keto PG F1 alpha was enhanced by 40 and 29% from macrophages preincubated with 10 microM curcumin or capsaicin, respectively, as compared to those produced by control cells. Curcumin and capsaicin also inhibited the secretion of collagenase, elastase, and hyaluronidase to the maximum extent of 57, 61, 66%, and 46, 69, 67%, respectively. These results demonstrated that curcumin and capsaicin can control the release of inflammatory mediators such as eicosanoids and hydrolytic enzymes secreted by macrophages and thereby may exhibit antiinflammatory properties. PMID- 9397404 TI - Generation and remodeling of highly polyunsaturated molecular species of rat hepatocyte phospholipids. AB - Freshly isolated rat hepatocytes were incubated for 20 min with [U-14C]glycerol in the presence or absence of unlabeled linoleic (18:2n-6), arachidonic (20:4n 6), or docosahexaenoic (22:6n-3) acid, added as albumin complex in 10% ethanol. Most of the radioactivity (approximately 95%) recovered in hepatocyte lipids was present in phosphatidylcholine (PC), phosphatidylethanolamine (PE), and triacylglycerol (TAG). The presence of exogenous fatty acids resulted in (i) higher incorporation of [U-14C]glycerol, (ii) higher percentage of label in TAG, and (iii) enhanced formation of PC and PE molecular species bearing the exogenous fatty acid at both the sn-1 and sn-2 positions of glycerol. In each case, these molecular species contained 60 to 70% of the label in that lipid class. Further incubation of the cells for 40 and 80 min in the absence of labeled substrate and exogenous fatty acids resulted in a redistribution of label among PC and PE molecular species due to deacylation-reacylation at the sn-1 position of glycerol. PMID- 9397405 TI - Analysis of the seed oil of Heisteria silvanii (Olacaceae)--a rich source of a novel C18 acetylenic fatty acid. AB - Besides some usual fatty acids (FA), two conjugated ene-yne acetylenic FA [trans 10-heptadecen-8-ynoic acid (pyrulic acid) (7.4%), and trans-11-octadecen-9-ynoic acid (ximenynic acid) (3.5%)], a novel ene-yne-ene acetylenic FA [cis-7, trans-11 octadecadiene-9-ynoic acid (heisteric acid) (22.6%)], and 9,10-epoxystearic acid (0.6%) could be identified in the seed oil of Heisteria silvanii (Olacaceae). Two further conjugated acetylenic FA [9,11-octadecadiynoic acid (0.1%) and 13 octadecene-9,11-diynoic acid (0.4%)] were identified tentatively by their mass spectra. The FA mixture has been analyzed by gas chromatography/mass spectrometry (GC/MS) of their methyl ester and 4,4-dimethyloxazoline derivatives. The structure of heisteric acid was elucidated after isolation via preparative silver ion thin-layer chromatography and by various spectroscopic methods [ultraviolet; infrared; 1H, 13C nuclear magnetic resonance (NMR); 1H-1H and 1H-13C correlation spectroscopy]. To determine the position of the conjugated ene-yne-ene system, the NMR spectra were also measured after addition of the lanthanide shift reagent Resolve-Al EuFOD. Furthermore, the triyglyceride mixture was analyzed by high temperature GC and high-temperature GC coupled with negative chemical ionization MS. A glass capillary column coated with a methoxy-terminated 50%-diphenyl-50% dimethylpolysiloxane was used for the separation of the triacylglycerol (TAG) species. No evidence of decomposition of the TAG species containing conjugated ene-yne-ene FA was observed. Twenty-six species of the separated TAG were identified by means of their abundant quasi molecular ion [M - H]- and their corresponding carboxylate anions [RCOO]- of the fatty acids, respectively. The major molecular species of the TAG were found to be 16:0/18:1/18:1, 16:0/18:1/18:3 (heisteric acid), 17:2 (pyrulic acid)/18:1/18:1, 18:1/18:1/18:3 (heisteric acid). The TAG containing acetylenic FA showed an unexpected increase of the retention time in comparison to the TAG containing usual FA, thus making the prediction of the elution order of lipid samples containing acetylenic FA difficult. PMID- 9397406 TI - Lipid specificity and location of the sterol carrier protein-2 fatty acid-binding site: a fluorescence displacement and energy transfer study. AB - Although it was recently recognized that sterol carrier protein-2 (SCP-2) interacts with fatty acids, little is known regarding the specificity of SCP-2 for long-chain fatty acids or branched-chain fatty-acid-like molecules. Likewise the location of the fatty-acid binding site within SCP-2 is unresolved. A fluorescent cis-parinaric acid displacement assay was used to show that SCP-2 optimally interacted with 14-22 carbon chain lipidic molecules: polyunsaturated fatty acids > monounsaturated, saturated > branched-chain isoprenoids > branched chain phytol-derived fatty acids. In contrast, the other major fatty-acid binding protein in liver, fatty-acid binding protein (L-FABP), displayed a much narrower carbon chain preference in general: polyunsaturated fatty acids > branched-chain phytol-derived fatty acids > 14- and 16-carbon saturated > branched-chain isoprenoids. However, both SCP-2 and L-FABP displayed a very similar unsaturated fatty-acid specificity profile. The presence and location of the SCP-2 lipid binding site were investigated by fluorescence energy transfer. The distance between the SCP-2 Trp50 and bound cis-parinaric acid was determined to be 40 A. Thus, the SCP-2 fatty-acid binding site appeared to be located on the opposite side of the SCP-2 Trp50. These findings not only contribute to our understanding of the SCP-2 ligand binding site but also provide evidence suggesting a potential role for SCP-2 and/or L-FABP in metabolism of branched-chain fatty acids and isoprenoids. PMID- 9397407 TI - Elution factors of synthetic oxotriacylglycerols as an aid in identification of peroxidized natural triacylglycerols by reverse-phase high-performance liquid chromatography with electrospray mass spectrometry. AB - Selected elution factors were determined for model oxotriacylglycerols as an aid in identification of the peroxidation products of natural triacylglycerols by reverse-phase high-performance liquid chromatography (HPLC) with electrospray mass spectrometry (LC/ES/MS). For this purpose synthetic triacylglycerols of known structure were converted to hydroperoxides, hydroxides, epoxides, and core aldehydes and their dinitrophenylhydrazones by published procedures. The oxotriacylglycerols were resolved by normal-phase thin-layer chromatography and reverse-phase HPLC, and the identities of the oxotriacylglycerols confirmed by LC/ES/MS. Elution factors of oxotriacylglycerols were determined in relation to a homologous series of saturated triacylglycerols, ranging from 24 to 54 acyl carbons, and analyzed by reverse-phase HPLC, using a gradient of 20-80% isopropanol in methanol as eluting solvent and an evaporative light-scattering detector. It was shown that the elution times varied with the nature of the functional group and its regiolocation in the triacylglycerol molecule. A total of 31 incremental elution factors were calculated from chromatography of 33 oxygenated and nonoxygenated triacylglycerol species, ranging in carbon number from 36 to 54 and in double-bond number from 0 to 6. PMID- 9397409 TI - Effects of high pressure and temperature on micelle formation of sodium deoxycholate and sodium dodecylsulfate. PMID- 9397408 TI - Evaluating acid and base catalysts in the methylation of milk and rumen fatty acids with special emphasis on conjugated dienes and total trans fatty acids. AB - Milk analysis is receiving increased attention. Milk contains conjugated octadecadienoic acids (18:2) purported to be anticarcinogenic, low levels of essential fatty acids, and trans fatty acids that increase when essential fatty acids are increased in dairy rations. Milk and rumen fatty acid methyl esters (FAME) were prepared using several acid- (HCl, BF3, acetyl chloride, H2SO4) or base-catalysts (NaOCH3, tetramethylguanidine, diazomethane), or combinations thereof. All acid-catalyzed procedures resulted in decreased cis/trans (delta 9c,11t-18:2) and increased trans/trans (delta 9t,11t-18:2) conjugated dienes and the production of allylic methoxy artifacts. The methoxy artifacts were identified by gas-liquid chromatography (Gl.C)-mass spectroscopy. The base catalyzed procedures gave no isomerization of conjugated dienes and no methoxy artifacts, but they did not transesterify N-acyl lipids such as sphingomyelin, and NaOCH3 did not methylate free fatty acids. In addition, reaction with tetramethylguanidine coextracted material with hexane that interfered with the determination of the short-chain FAME by GLC. Acid-catalyzed methylation resulted in the loss of about 12% total conjugated dienes, 42% recovery of the delta 9c,11t-18:2 isomer, a fourfold increase in delta 9t,11t-18:2, and the formation of methoxy artifacts, compared with the base-catalyzed reactions. Total milk FAME showed significant infrared (IR) absorption due to conjugated dienes at 985 and 948 cm-1. The IR determination of total trans content of milk FAME was not fully satisfactory because the 966 cm-1 trans band overlapped with the conjugated diene bands. IR accuracy was limited by the fact that the absorptivity of methyl elaidate, used as calibration standard, was different from those of the other minor trans fatty acids (e.g., dienes) found in milk. In addition, acid-catalyzed reactions produced interfering material that absorbed extensively in the trans IR region. No single method or combination of methods could adequately prepare FAME from all lipid classes in milk or rumen lipids, and not affect the conjugated dienes. The best compromise for milk fatty acids was obtained with NaOCH3 followed by HCl or BF3, or diazomethane followed by NaOCH3, being aware that sphingomyelins are ignored. For rumen samples, the best method was diazomethane followed by NaOCH3. PMID- 9397410 TI - Isomers in commercial samples of conjugated linoleic acid. PMID- 9397411 TI - Determinants of protein turnover in health and disease. AB - Protein synthesis, protein degradation, and amino acid oxidation are tightly regulated to preserve lean body mass in healthy individuals. An adaptative response to a reduction in dietary protein in normal adults is decreased branched chain amino acid oxidation which increases the availability of amino acids. In nephrosis, reduced branched-chain amino acid oxidation decreases amino acid requirements and helps to compensate for urinary protein loss. Conversely, uremia and other catabolic diseases are associated with muscle wasting resulting from activation of the ubiquitin-proteasome proteolytic pathway and branched-chain ketoacid dehydrogenase, the rate-limiting enzyme for branched-chain amino acid catabolism. By understanding the processes responsible for muscle wasting in catabolic states, therapeutic interventions may be designed to improve protein balance. PMID- 9397412 TI - Metabolic acidosis and protein catabolism: mechanisms and clinical implications. AB - Metabolic acidosis increases protein degradation resulting in muscle wasting and a negative nitrogen balance. The branched-chain amino acids serve as useful markers of these changes and their catabolism is increased in acidosis, particularly for the spontaneous acidosis associated with renal failure. As a result, the neutral nitrogen balance is compromised and malnutrition results. Glucocorticoids mediate these changes through the recently discovered ATP dependent ubiquitin-proteasome pathway. Therapy necessitates correction of the underlying acidosis either through adjustment of the alkalinity of the dialysate for the patient on dialysis or through dietary protein restriction and sodium bicarbonate supplements for the predialysis patient. PMID- 9397413 TI - The impact of malnutrition on kidney function. AB - Malnutrition is the most common cause of mortality in the world. It affects underdeveloped as well as industrialized societies, in the latter demonstrating a prevalence in hospitalized patients of between 30 and 50%. Although the prevalence has decreased in recent studies, the problem is still significant among a selected group of patients. The clinical manifestations of malnutrition may be evident on physical examination but alterations in renal function may not show up at the initial exam. Clinical and experimental models of protein-calorie malnutrition have confirmed significant alterations in renal hemodynamics, renal concentration capacity, and renal acid excretion. Children and adults with malnutrition have been shown to have a decreased glomerular filtration rate and renal plasma flow (RPF), as well as a lowered capacity to concentrate the urine and excrete an acid load. Moreover, clinical and experimental models of protein calorie malnutrition have unravelled the roles of the renin-angiotensin system, renal prostaglandins, and urea production in the renal function changes associated with malnutrition. We have reviewed the most pertinent and recent studies from our and other laboratories which have improved our understanding of renal functional alterations in malnutrition. PMID- 9397414 TI - Growth factors: future prospects in renal failure. AB - Chronic renal failure is associated with an abnormal growth hormone/insulin-like growth factor-1 axis. In addition, nutritional status strongly regulates this axis. Because these hormones are involved in growth in children and maintenance of a normal body composition in adults, experimental and clinical studies have tested the metabolic effects of these recombinant growth factors. Various conditions in which these growth factors have been administered have been reported, such as the recovery of acute renal failure, protein metabolism in chronic renal failure, growth improvement in uremic children, the increase in renal function in nondialyzed uremic patients, and the potential treatment of malnutrition in adult maintenance dialysis patients. PMID- 9397415 TI - Impact of chronic renal failure on nitrogen metabolism. AB - Evidence indicates that both nephrotic and nonnephrotic chronic renal failure (CRF) patients can activate normal compensatory responses when dietary protein intake is restricted and that their protein and energy requirements are similar to normal subjects. When properly implemented, low-protein diets are safe and the benefits include the amelioration of uremic symptoms and some of their metabolic complications and possibly a reduction in the rate of progression of renal failure. To ensure dietary adequacy and compliance, patients should be monitored when treated with low-protein diets. Recent evidence that the protein intake of patients with progressive CRF declines when they consume unrestricted diets should not be considered as an argument against the use of low-protein diets. Rather, it is a persuasive argument in favor of restricting dietary protein intake to minimize the complications of renal failure. PMID- 9397416 TI - Derangements in protein metabolism induced by type I diabetes mellitus. AB - In poorly controlled diabetics, the whole-body protein flux is increased by 20 30% in comparison to well-controlled type I diabetes (IDDM) and normal subjects. Intensive insulin administration completely reverses these abnormalities. In poorly controlled IDDM, the primary effect of insulin administration is to reduce the increased protein catabolic rate by suppressing the accelerated rate of protein breakdown. Studies in humans have demonstrated that the increased rate of protein synthesis observed in these patients is the consequence of elevated plasma amino acid levels. When IDDM subjects develop renal complications, a protein-restricted diet may be recommended to preserve the remnant kidney function. However, it has been demonstrated that in IDDM patients, metabolic adaptation to protein restriction is incomplete because suppression of endogenous proteolysis is impaired. Since this component of protein metabolism is very sensitive to insulin action, maintaining strict metabolic control during the protein restriction regimen has been suggested. The major studies on the effects of amino acid and insulin on protein metabolism in patients with diabetes mellitus are reviewed. PMID- 9397417 TI - Protein metabolism in acute renal failure. AB - The hallmark of metabolic alterations in acute renal failure (ARF) is accelerated protein breakdown which, unfortunately, cannot be suppressed effectively by provision of exogenous nutritional substrates. Causes of excessive protein catabolism are manifold and present a combination of unspecific mechanisms induced by the acute disease process and underlying illness or associated complications, effects induced by the acute loss of renal function and, finally, the type and intensity of renal replacement therapy. Specific uremic toxic effects, insulin resistance, hormonal derangements, metabolic acidosis, circulating proteases, inflammatory mediators, and dialysis-related losses of nutritional substrates all contribute to the activation of protein degradation. Metabolism in ARF is also affected by an impairment of the multiple metabolic and endocrine functions of the kidney. Various amino acids are synthesized or interconverted by the kidneys and may become conditionally indispensable. The kidney is also an important organ in the degradation of peptides, such as peptide hormones. As a consequence of these metabolic aberrations, imbalances in amino acid pools in plasma and in the intracellular compartment occur in ARF and elimination and utilization of infused amino acids is altered. Protein or amino acids requirements are influenced more by the nature of the illness causing ARF, by the extent of hypercatabolism, by associated complications, and by the type and frequency of renal replacement therapy than by renal dysfunction per se. In noncatabolic patients, an intake of 1 g/kg body weight per day may be sufficient, while in critically ill hypercatabolic patients undergoing continuous renal replacement therapy, 1.5 g amino acids/kg body weight per day should be provided to minimize nitrogen losses. For the future, we need to identify safe methods to control the accelerated catabolism in order to improve the efficiency of nutritional interventions in patients with ARF. PMID- 9397418 TI - Albumin turnover in renal disease. AB - Hypoalbuminemia is found in patients both with the nephrotic syndrome and with end-stage renal disease (ESRD) treated either with continuous ambulatory peritoneal dialysis (CAPD) or hemodialysis. In nephrotic patients the primary causes of hypoalbuminemia are urinary albumin losses, an inappropriate increase in the fractional catabolic rate (FCR) of albumin and an insufficient increase in the rate of albumin synthesis to replace these losses. Nevertheless, the albumin synthetic rate is increased significantly. In patients on CAPD, albumin losses into the urine and across the peritoneal membrane contribute significantly to hypoalbuminemia. In contrast to nephrotic patients, albumin FCR decreases as serum albumin falls and serum albumin levels are significantly greater than in nephrotic patients with the same external losses of albumin. CAPD patients, like nephrotic patients with normal renal function, can increase albumin synthesis to replace losses. Thus ESRD does not directly suppress albumin synthesis. In contrast, hypoalbuminemia in hemodialysis patients results from reduced albumin synthesis. The cause of decreased albumin synthesis is a combination of response to inflammation (acute-phase response) and, to a lesser extent, inadequate nutrition. There is no evidence that shifts of albumin to the extravascular space or that dilution of the plasma by volume expansion play any role in causing hypoalbuminemia in ESRD or nephrotic patients. PMID- 9397419 TI - Effects of a supplemented very low protein diet in predialysis patients on the serum albumin level, proteinuria, and subsequent survival on dialysis. AB - A very low protein diet (0.3 g/kg ideal body weight) supplemented with essential amino acids (or ketoanalogues) is seldom employed at present in chronic renal failure for fear of inducing protein deficiency, especially in patients who also have the nephrotic syndrome. Nevertheless, we have used this dietary regimen in predialysis patients for a number of years. We have shown that when these patients reach the end stage, they rarely exhibit hypoalbuminemia, in contrast to the reported 25-50% hypoalbuminemia at the onset of dialysis nationwide. Furthermore, their survival for the first 2 years on dialysis is much improved, in comparison with the national experience, adjusted for age, sex, and cause of renal disease. When nephrotic patients are given this regimen, they exhibit some improvement in parameters of the nephrotic state, but nevertheless progress to dialysis, provided their initial glomerular filtration rate (GFR) is < 30 ml/min. However, if their initial GFR is > 30 ml/min, they may show gradual but complete remission of the nephrotic syndrome, even when the underlying disease is diabetic nephropathy or focal segmental glomerulosclerosis. We conclude that this dietary regimen is not only safe in patients with renal failure, with or without the nephrotic syndrome, but may be of substantial benefit. The mechanism remains to be explained. PMID- 9397420 TI - Nutritional status and survival in end-stage renal disease patients. AB - Several reports have emphasized that putative laboratory surrogates of nutrition, such as serum albumin, creatinine, and cholesterol concentrations are statistically more powerful independent predictors of odds risk of death for dialysis patients than is the delivered dose of dialysis. In view of the relative simplicity with which these blood tests can be obtained, their lack of expense, and simplicity in interpretation, the dialysis community has greatly escalated their importance as performance measures for the processes of patient care, arguably without full consideration of their meaning. If malnutrition in dialysis patients is a powerful predictor of death risk, and is amenable to corrective interventions that result in a reduction in the odds risk of death, then the zeal with which these laboratory tests have been embraced is appropriate. However, the assumption that a statistical correlation between laboratory surrogates of malnutrition, or other measures of inadequate nutrition, such as body mass index or a subjective global assessment, indicate a direct causal relationship between nutritional intake, nutritional status, and outcome may be incorrect. Such apparent linkages may be a consequence of the statistical model selected alone, i.e., another unappreciated medical condition may be the proximate cause of death in addition to resulting in malnutrition. The mechanism(s) by which malnutrition may adversely impact the survival of end-stage renal disease (ESRD) patients is unclear. The impact of milder degrees of malnutrition on patient survival, their proximate effect on survival, and the reality of their independent effect on patient survival are also inadequately defined. Clearly, there is a statistical link between the putative laboratory surrogates of nutrition and patient survival. Regardless of the pathobiology of such a causal link, it is valid to enquire if an intervention that results in a positive change in nutritional parameters enhances patient survival. These issues surrounding nutritional status and survival in patients with ESRD are reviewed here in detail. The conclusion of this critique is that additional studies are needed to determine if malnutrition is truly an independent and responsive predictor of outcome for ESRD patients. PMID- 9397421 TI - Strategies for nutritional intervention in patients with renal failure. AB - This review article discusses the evidence documenting the interrelationship between nutritional status and clinical outcome of the renal patient population. The limitations of accurately assessing the nutritional status of this patient group with commonly used indices are detailed. An overview of the history of nutrition supplementation as an intervention for the malnourished renal patient reveals that the efficacy of both enteral and parenteral methods has not been adequately explored to draw any firm conclusions. Based on available data, recommendations for providing nutrition care for the malnourished patient are provided. Therapeutics that are currently under investigation as future interventions for malnutrition are identified. PMID- 9397422 TI - Factors contributing to catabolism in end-stage renal disease patients. AB - End-stage renal disease (ESRD) patients, whether they are treated with hemodialysis or continuous ambulatory peritoneal dialysis, frequently suffer from protein-energy malnutrition, which is associated with increased morbidity and mortality. The protein requirements in dialysis patients are increased compared to those of healthy individuals and nondialyzed patients with chronic renal failure. The intake of protein and energy is frequently reduced because of the underlying disease, comorbidity, psychosocial factors, and uremic anorexia (underdialysis). There are several factors in ESRD patients that may enhance protein catabolism and increase protein requirements, such as low energy intake, amino acid abnormalities, metabolic acidosis, endocrine abnormalities (insulin resistance, hyperglucagonemia, hyperparathyroidism, insensitivity to growth hormone and insulin-like growth factor-1, cardiac failure, infection and inflammation, anemia, and physical inactivity. The dialytic procedures per se may enhance protein catabolism due to dialytic losses of protein and amino acids and, in hemodialysis, an inflammatory response to blood-dialyzer interaction. The relative importance of the various factors which cause anorexia and stimulate protein catabolism is still not well understood. PMID- 9397423 TI - Motivational properties of oxytocin in the conditioned place preference paradigm. AB - We hypothesized that oxytocin might have intrinsic reinforcing properties and studied it using a conditioned place preference. Three studies examining motivational properties of oxytocin in nonpreferred, preferred, and balance designs were performed utilizing two compartment apparatus. On alternate days, compartments were paired with subcutaneously injected oxytocin (6 mg/kg) or saline, and animal pre- and post-conditioning place preference was compared. Whereas in animals paired with saline there was a shift to a lack of preference, oxytocin-treated animals reversed their preference, spending more time in a previously unpreferred, compartment. In preferred compartment design, oxytocin treated animals further increased their preference, whereas saline-treated animals decreased their preference toward a nonpreference for either compartment. Our results demonstrate that oxytocin produces a reliable and robust preference for the environment with which it is repeatedly associated, and has rewarding or potentially anti-aversive properties. Future studies are needed to distinguish among these possibilities. PMID- 9397424 TI - Effects of the 5-HT3 antagonist, ondansetron, on the behavioral and physiological effects of pentagastrin in patients with panic disorder and social phobia. AB - Pentagastrin, a cholecystokinin (CCK) agonist, produces anxiety and panic in patients with panic disorder and social phobia. Preclinical data suggests that pentagastrin-induced anxiogenesis may be mediated via 5-HT3 receptors. In the present study, 14 patients with panic disorder or social phobia underwent pharmacological challenge in three conditions: (1) pretreatment with saline followed by pentagastrin infusion; (2) pretreatment with ondansetron followed by pentagastrin infusion; and (3) pretreatment with saline followed by saline infusion. As expected, pentagastrin administration led to increased anxiety, physical symptoms of panic attacks, pulse, plasma adrenocorticotropic hormone (ACTH), and cortisol. Pentagastrin's behavioral effects were not blocked by ondansetron, and in fact, tended to be exaggerated. Ondansetron pretreatment did not alter the pentagastrin-induced cortisol increase but significantly prolonged the pentagastrin-induced increase in ACTH. These findings suggest that pentagastrin's behavioral effects are not mediated by 5HT3 receptors. Mechanisms by which peripherally administered CCK agonists lead to anxiety remain to be elucidated. PMID- 9397425 TI - Patients with premenstrual syndrome have reduced sensitivity to midazolam compared to control subjects. AB - Premenstrual syndrome (PMS) depends on gonadal hormones produced by the corpus luteum. Given the facilitory actions on GABAergic inhibitory neurotransmission exerted by certain progesterone metabolites, further studies on the GABAA receptor system in premenstrual syndrome are warranted. This study evaluated the benzodiazepine sensitivity in PMS patients and control subjects, using saccadic eye velocity (SEV) and visual analogue ratings of sedation as dependent measures. PMS patients displayed a significantly reduced SEV responsiveness to benzodiazepines compared to control subjects in the follicular phase, whereas there was no difference between groups in the luteal phase. In the luteal phase, the sedation response to benzodiazepines was significantly reduced in PMS patients compared to control subjects. There was also an influence of PMS symptom severity on these measures, as high-severity PMS patients displayed blunted SEV and sedation responses to benzodiazepines compared to low-severity patients. These results indicate that PMS patients have a reduced functional sensitivity at the GABAA/benzodiazepine receptor complex throughout the menstrual cycle. PMID- 9397426 TI - Effects of divalproex sodium on 5-HT1A receptor function in healthy human males: hypothermic, hormonal, and behavioral responses to ipsapirone. AB - Hypothermic and hormonal responses to a challenge with a selective 5-HT1A receptor agonist ipsapirone are considered to provide an index of 5-HT1A receptor function in humans. To examine the effects of divalproex sodium (DVP) on 5-HT1A receptor function in humans, we measured the hypothermic, adrenocorticotropic hormone (ACTH) cortisol, and behavioral responses to ipsapirone in 10 healthy male volunteers. After obtaining a blood sample for baseline hormone levels and measuring body temperature, a single dose of 0.3 mg/kg of ipsapirone was given orally to all the subjects and further bloods and temperature reading were obtained at regular intervals for three hours. The ipsapirone challenge tests were repeated after the subjects had been treated with DVP (1000 mg/day) for one week. The results showed that the hypothermia induced by ipsapirone was significantly attenuated by the DVP treatment, whereas the ACTH/cortisol release and the behavioral responses following ipsapirone challenges were not altered. Our findings suggest that DVP may enhance 5-HT neurotransmission in humans via a subsensitization of 5-HT1A autoreceptors but does not appear to affect postsynaptic 5-HT1A receptors. PMID- 9397427 TI - Intravenous dextroamphetamine and brain glucose metabolism. AB - This study reports the effects of intravenous dextroamphetamine on cerebral glucose metabolism assayed by positron emission tomography (PET) and [fluorine 18]fluorodeoxyglucose (FDG) in 13 healthy adults during the performance of a continuous visual attention task. Two FDG PET scans were performed within a single experimental session. The first scan was preceded by the injection of placebo and the second scan by the injection of 0.15 mg/kg dextroamphetamine. Global and normalized regional glucose metabolic rates (rCMRglc) were examined as a function of pharmacological challenge and subjective experience. Subcortical, limbic, frontal, and cerebellar rCMRglc significantly increased after dextroamphetamine, whereas rCMRglc of the temporal cortex significantly decreased. Physiological and self-report measures of subjective states showed the expected alterations. These rCMRglc changes reflect both the direct pharmacological effect of dextroamphetamine on monoaminergic neurotransmitter systems as well as enhancement of the activation of the neural network mediating the performance of the continuous attention task. PMID- 9397428 TI - Decreasing striatal 6-FDOPA uptake with increasing duration of cocaine withdrawal. AB - It has been hypothesized that a decrease in dopaminergic presynaptic activity during abstinence or withdrawal is related to relapse in cocaine-dependent subjects (Dackis and Gold 1985; Markou and Koob 1991). This study measured striatal 6-fluorodopa (6-FDOPA) uptake, an index of dopaminergic presynaptic activity, using positron emission tomography (PET) in 11 drug-free cocaine addicts compared to eight normal subjects. Middle abstinence cocaine addicts (n = 5, off cocaine 11-30 days) had significantly lower striatal 6-FDOPA uptake compared to normal controls or early abstinence cocaine addicts (n = 6, off cocaine 1-10 days). The cocaine-dependent subjects (n = 11) showed a significant negative correlation between days off cocaine and striatal 6-FDOPA uptake. The results suggest that during abstinence from cocaine there is a delayed decrease in dopamine terminal activity in the striatum. PMID- 9397429 TI - The rise and fall of HMOs. PMID- 9397430 TI - The clinical utility of duplicate readings for musculoskeletal radiographs. AB - It is a common practice in many hospitals to have all skeletal radiographs read by a second physician, usually a radiologist, as well as by the treating physician. A two-part study was performed in order to examine the cost and clinical benefit of this practice for plain films ordered by orthopedists. In the first part of this study, the attending orthopedic surgeons were surveyed about the clinical usefulness and effect on patient care of 1000 radiologic reports from plain films ordered on orthopedic patients. In the second part, the charts of 272 patients who had 704 radiographs were reviewed with the goal of identifying any discrepancies between the orthopedic interpretation and the radiologic reading. Thirty-eight reports were discarded because they were not reports of plain skeletal films. One hundred twenty-nine of the remaining 962 radiologic reports were never read by the attending orthopedist. The average time between the taking of the film and an orthopedic attending reading the printed report was 6.1 days. Three radiology reports contained findings that were incorrect. Only one report contained findings that the orthopedist was unaware, and one report may have led to an alteration in treatment. No reports resulted in an unplanned trip to the operating room or a patient being called back to the clinic. Of the 272 chart reviews (704 reports), 70 had no orthopedic interpretation recorded and 94 had no radiologic report in the chart. Twelve discrepancies were noted in the cases that had both reports. Four fracture displacements were identified by orthopedists, but not on the written radiology report; three of these required a return to the operating room. Four instances of hardware displacement or breakage were noted by orthopedists, but not commented on by the radiologists. Three incidental injuries (two fractures and an acromioclavicular injury) were noted on printed reports of films taken for other reasons, but not commented on by the orthopedist, and not treated. A dorsal bunion was noted on one film by the orthopedist, but not by the radiologist. From this study, one can conclude that the benefit of routine duplicate radiograph interpretation by a second physician does not justify its cost. PMID- 9397431 TI - Clinical results of the modular porous-coated anatomic (PCA) total knee arthroplasty with cement: a 5-year prospective study. AB - Our study examines the clinical, radiographic, and patient satisfaction outcome of the cemented Modular Porous-Coated Anatomic (PCA) total knee arthroplasty with a minimum 5-year follow up. All data were gathered prospectively and consecutively. Patient satisfaction was assessed with a self-administered survey. Statistical analysis examined the effect of 17 patient factors, 19 surgical factors, and postoperative continuous passive motion use on range of motion (ROM) and HSS scores at 2 years. Seventy-eight Modular PCA arthroplasties performed by 9 orthopedic surgeons on 71 patients between January 1988 and November 1989 are reported in this study. Preoperative HSS scores averaged 51.2 and improved to an average of 89 at 1 and 2 years, and 86 at 5 years after surgery (90% good or excellent). ROM changed after surgery through improvement in preoperative knee flexion contracture, but not in increased knee flexion. One patient underwent reoperation for patellar instability, and one patient's arthroplasty was revised at 53 months for late instability. The total reoperation rate for any reason was 7.7%. Zonal analysis for progressive radiolucency at the bone-cement interface showed increasing frequency of narrow (< 1 mm) radiolucencies concentrated on the anterior and medial aspect of the tibial tray. Ninety-eight percent of patients responded to an outcome questionnaire, and 96% rated themselves improved. The Kaplan-Meier probability of an implant surviving without loosening at 5 years was 100%. The Modular PCA TKA has a low incidence of patellofemoral problems, is clinically successful, and results are stable at a minimum 5-year follow-up examination. PMID- 9397432 TI - Freeze-dried cortical allograft in posterior spinal arthrodesis: use with segmental instrumentation for idiopathic adolescent scoliosis. AB - Many surgeons use cancellous or corticocancellous allogeneic bone grafts to augment spinal arthrodeses. The efficacy of onlay freeze-dried cortical allografts for posterior spinal fusions has been questioned in the past. In this report, freeze-dried, crushed cortical bone allograft was used as the sole fusion material in 32 consecutive posterior spinal fusions for idiopathic adolescent scoliosis. Absence of clinical or radiographic pseudarthrosis in all patients at an average follow up of 34 months suggests that freeze-dried, crushed cortical bone allograft is a suitable material for augmentation of the fusion mass during posterior spinal fusion with segmental instrumentation. PMID- 9397433 TI - Repair of soft tissue to bone using a biodegradable suture anchor. AB - The medial collateral ligaments of 18 New Zealand rabbits were surgically detached from bone. In one knee, the ligament was repaired using a biodegradable suture anchor composed of a co-polymer of lactic and glycolic acid. The contralateral medial collateral ligament was not repaired. Animals were sacrificed at 4, 8, and 12 weeks after the operation, and the knee that had the ligament repair was compared with the contralateral control knee. All knees were tested manually tested for stability to valgus stress and then prepared for histologic examination. Medial collateral ligaments repaired using the biodegradable suture anchor demonstrated stability to valgus stress and anatomic healing at the bone-tendon junction. Resorption of the implant was virtually complete by 12 weeks. All specimens demonstrated less inflammatory reaction to the suture anchor than to the attached Vicryl suture. This contrasts with the control group, which was grossly unstable and demonstrated scarring in this nonanatomic position. These results demonstrate efficacy of this particular material of biodegradable implant and justify further investigative efforts. PMID- 9397434 TI - Limited range of motion after total knee arthroplasty: etiology, treatment, and prognosis. PMID- 9397435 TI - Percutaneous pinning of proximal humerus fractures: a biomechanical study. AB - Mechanical testing of two-part surgical neck fractures fixed with four different pin configurations was performed. Ten fresh, frozen, unembalmed humeri stripped of all soft tissues were used; the surgical neck was osteotomized perpendicular to the humerus long axis. Terminally threaded 2.5-mm AO pins were used to fix the fracture. Humeri then were tested in both torsion and bending on a custom-made jig using Instron 1331 to assess the rigidity of pinning constructs. In torsion, two lateral pin construct was significantly less rigid than all other pin configurations. The addition of an anterior pin to two lateral pins did not increase bending rigidity, but significantly increased torsional stiffness. The addition of two bicortical tuberosity pins or two bicortical tuberosity pins and one anterior pin to two lateral pins significantly increased rotational and bending rigidity. Results confirm clinical data, and the authors conclude that multiplanar pins are needed to augment torsional stiffness, and that the addition of two bicortical tuberosity pins enhances bending rigidity. PMID- 9397436 TI - Treatment of isolated fractures of the ulnar shaft. PMID- 9397437 TI - Intraosseous gout: an "aggressive" solitary lesion. PMID- 9397438 TI - Pseudoaneurysm following femoral fracture. AB - If the physician is aware of this diagnosis and maintains an appropriate level of suspicion and low threshold to commence duplex evaluation. The potential morbidity of a fracture-induced traumatic pseudoaneurysm can be minimized. PMID- 9397439 TI - Removal of a broken solid-core intramedullary femoral nail using both antegrade and retrograde starting points. PMID- 9397440 TI - Traumatic rupture of the transverse carpal ligament associated with compartment syndrome of the hand. PMID- 9397441 TI - Radiologic case study. Pes anserine bursitis. PMID- 9397442 TI - On patient compliance with medication doses. PMID- 9397443 TI - A pediatrician's view. A goal-oriented approach to managing children with special healthcare needs. PMID- 9397444 TI - Profiling the health service needs of populations: description and uses of the NACHRI Classification of Congenital and Chronic Health Conditions. PMID- 9397445 TI - Pediatricians partnering with states to assure that children with special health needs are provided appropriate services: the Vermont experience with managed Medicaid. PMID- 9397446 TI - Managed care for children with special healthcare needs: Michigan's approach. PMID- 9397447 TI - Paradigms of care for children with special healthcare needs. PMID- 9397448 TI - Community resources for children with special healthcare needs. PMID- 9397449 TI - The care of children with chronic illness in primary care practice: implications for the pediatric generalist. PMID- 9397450 TI - Bronchioloalveolar carcinoma: clinical, histopathologic, and radiologic findings. AB - Bronchioloalveolar carcinoma is characterized pathologically by a pulmonary neoplasm showing lepidic growth. More than half of all patients with bronchioloalveolar carcinoma are asymptomatic. The most frequent symptoms and signs are cough, sputum, shortness of breath, weight loss, hemoptysis, and fever. Bronchorrhea is unusual and a late manifestation. Nonmucinous bronchioloalveolar carcinoma tends to be more localized and has a lower frequency of bronchogenic spread than mucinous bronchioloalveolar carcinoma. Bronchioloalveolar carcinoma appears radiographically as a single nodule, segmental or lobar consolidation, or diffuse nodules. At computed tomography (CT), the single nodular form appears as a peripheral nodule or localized ground-glass attenuation with or without consolidation, frequently associated with bubblelike areas of low attenuation and open bronchus signs. The lobar consolidative form may demonstrate the CT angiogram and open bronchus signs. The diffuse nodular form appears as multiple nodules or areas of ground-glass attenuation or consolidation. The single nodular form has a better prognosis than the others but may show false-negative results for malignancy at 2-(fluorine-18) fluoro-2-deoxy-D-glucose positron emission tomography. PMID- 9397451 TI - Pulmonary complications after bone marrow transplantation: high-resolution CT and pathologic findings. AB - A wide variety of pulmonary complications occur in bone marrow transplant (BMT) recipients and are a major cause of morbidity and death. High-resolution computed tomography (CT) is excellent in the detection of pulmonary abnormalities, but these findings are generally nonspecific. However, the different complications, which reflect the immunologic status of the patients, occur in three phases. This pattern can be used to interpret CT scans. The neutropenic phase (up to 3 weeks after BMT) is characterized by fungal infections, notably angioinvasive aspergillosis, alveolar hemorrhage, pulmonary edema, and drug reactions. At CT, angioinvasive aspergillosis appears as a nodule surrounded by a halo of ground glass attenuation; alveolar hemorrhage and drug reactions, as bilateral areas of ground-glass attenuation or consolidation; and pulmonary edema, as prominent pulmonary vessels, interlobar septal thickening, ground-glass attenuation, and pleural effusions. The second phase (3 weeks to 100 days after BMT) is dominated by cytomegalovirus pneumonia, which appears as multiple small nodules with associated areas of consolidation or ground-glass attenuation, and Pneumocystis carinii pneumonia, which appears predominantly as ground-glass attenuation. The late phase (more than 100 days after BMT) is characterized by bronchiolitis obliterans, bronchiolitis obliterans with organizing pneumonia (BOOP), and chronic graft-versus-host disease. In bronchiolitis obliterans, CT reveals bronchial dilatation and a mosaic pattern of attenuation; in BOOP, CT findings usually consist of patchy consolidation or ground-glass attenuation. If CT findings are considered in relation to the time elapsed after BMT, diagnostic options can be narrowed sufficiently to enable accurate diagnosis. PMID- 9397452 TI - Imaging and pathologic features of myelolipoma. AB - Myelolipoma is a benign tumor consisting of mature fat interspersed with hematopoietic elements resembling bone marrow. Imaging findings in a large series of pathologically proved cases of myelolipoma were correlated with the pathologic and histologic features of the lesions. Myelolipoma manifests in four distinct clinicopathologic patterns: isolated adrenal myelolipoma, adrenal myelolipoma with hemorrhage, extraadrenal myelolipoma, and myelolipoma associated with other adrenal disease. Myelolipoma is difficult or impossible to detect at plain radiography unless the lesion is large and predominantly fatty. At ultrasound, myelolipoma often has heterogeneous echogenicity due to its typically nonuniform architecture. Computed tomography (CT) frequently demonstrates large amounts of fat with areas of interspersed higher-attenuation tissue. At magnetic resonance imaging, predominantly fatty areas usually have increased signal intensity on T1 weighted images and moderate hyperintensity complicated by the presence of marrowlike elements in the corresponding regions on T2-weighted images. The imaging appearance of myelolipoma is altered by the presence of hemorrhage. In such cases, CT is the most accurate method for evaluation. Knowledge of the imaging characteristics of myelolipoma usually allows presumptive diagnosis, although percutaneous needle biopsy may be needed to confirm the diagnosis in cases of extraadrenal myelolipoma. Surgical excision is unnecessary unless the diagnosis is unclear or the lesion is symptomatic. Asymptomatic, nonhemorrhagic myelolipomas do not require therapy. PMID- 9397453 TI - In vitro and in vivo MR imaging of hyaline cartilage: zonal anatomy, imaging pitfalls, and pathologic conditions. AB - Hyaline cartilage plays an essential role in the maintenance of normal synovial joint function by reducing friction and distributing loads. Histologic analysis of hyaline cartilage reveals zonal variation in cellular morphology, proteoglycan concentration, and collagen fiber size and orientation. High-resolution magnetic resonance (MR) imaging reveals an analogous laminar anatomy that is often visible on clinical images obtained with proper attention to technique. In vitro and in vivo pulse sequences show three distinct laminae: a hypointense superficial lamina, a hyperintense intermediate lamina, and a heterogeneous deep lamina that consists of alternating hyperintense and hypointense bands perpendicular to the subchondral bone. Imaging pitfalls include magic angle effects, truncation artifact, partial volume effect, regional anatomic variation, chemical shift, and magnetic susceptibility effects. Pathologic conditions that affect articular cartilage include chondromalacia patellae, osteoarthritis, and localized traumatic lesions. Although detection of early cartilage disease remains elusive, MR imaging can demonstrate intermediate and advanced lesions. PMID- 9397454 TI - MR arthrography of the shoulder: variants and pitfalls. AB - Use of magnetic resonance arthrography to evaluate pathologic conditions of the shoulder is becoming widespread. However, normal anatomy or anatomic variations can cause interpretive errors. The most common variations occur at the origins of the glenohumeral ligaments (GHLs) and the insertion of the joint capsule. Among the GHL variants, common origin of the superior and middle ligaments is the most frequent followed by thinning, thickening, or absence of a ligament, most often the middle one. Absence or thinning of one ligament is sometimes associated with thickening of another or changes in the size and shape of the anterior capsular recesses. Common normal variants of the labrum include foramen sublabrum (detachment of the anterosuperior labrum from the glenoid margin) and the Buford complex (absence of the anterosuperior labrum in association with a thick middle GHL). Pitfalls related to the arthrographic technique include (a) visualization of a deep sulcus between the insertion of the long head of the biceps tendon and the superior labrum and (b) an apparent type III capsular insertion due to overdistention of the capsule by injected contrast material. PMID- 9397455 TI - Characterization of atherosclerotic plaques at the carotid bifurcation: correlation of high-resolution MR imaging with histologic analysis--preliminary study. AB - The clinical symptoms and morbidity that result from carotid artery disease, the primary cause of stroke, are mainly due to plaque ulceration, thrombosis, intraplaque hemorrhage, and thinned fibrous caps. The contents of atherosclerotic plaques of the carotid artery can be determined with in vivo high-resolution magnetic resonance imaging with flow suppression. Eight patients scheduled to undergo endarterectomy and four healthy volunteers were imaged with a 1.5-T imager and custom-made carotid phased-array coils. T1-weighted spin-echo images and cardiac-gated proton-density--weighted fast spin-echo images were acquired. In vivo imaging findings as determined by three radiologists were correlated with ex vivo imaging and histologic findings. Among the eight plaque specimens, regions of hemorrhage, calcium, lipid deposits, and fibrous plaques were identified on T1- and proton-density-weighted images. Calcium and lipid deposits were detectable on both T1- and proton-density--weighted images. Hemorrhage and fibrous plaques were better demonstrated on proton-density--weighted images. PMID- 9397456 TI - MR angiography of the portal venous system: techniques, interpretation, and clinical applications. AB - Magnetic resonance (MR) angiography is a noninvasive means of assessing the portal venous system that has potential advantages over currently used modalities. Time-of-flight and phase-contrast MR angiography are useful techniques that differ fundamentally in their means of data acquisition but are comparable in their ability to demonstrate normal anatomy as well as abnormalities of the portal venous system. Occasionally, artifacts caused by respiratory motion, implanted metallic devices or surgical clips, in-plane saturation, or areas of complex flow are seen at MR angiography of the portal venous system. However, most artifacts can easily be identified as such and either remedied or ignored. In addition, the suppression of signal from surrounding soft tissues may result in poor detection of parenchymal lesions. The utility of standard projection angiograms and source images can be increased through the use of intravenously administered contrast material and postprocessing techniques such as partial-volume maximum intensity projection reconstructions and shaded surface renderings. In addition to providing information on portal venous anatomy and portosystemic collateral vessels, MR angiography of the portal vein has clinical application in portal venous thrombosis and stenosis, liver transplantation, and the evaluation and planning of surgical and transjugular intrahepatic portosystemic shunts. PMID- 9397457 TI - CT appearances of ossicular injuries. AB - Trauma of the ossicular chain is a frequent complication of temporal bone injury. Skull trauma from blows to the temporal, parietal, or occipital region (with or without fracture of the temporal bone) is the main cause of ossicular injury; other modes of injury are rare. Ossicular injury usually occurs as a dislocation, of which there are five types: incudostapedial joint separation, incudomalleolar joint separation, dislocation of the incus, dislocation of the malleoincudal complex, and stapediovestibular dislocation. Fracture of the malleus, incus, or stapes is uncommon. High-resolution computed tomography is the method of choice for evaluation of ossicular trauma. Joint separation and fracture of the stapes are seen on axial images; coronal images may aid visualization. Both axial and coronal images are needed for evaluation of a dislocated malleus or incus. Fracture of the malleus or incus is detected with axial or coronal images; reformatted images may also be useful. PMID- 9397458 TI - Imaging features of radiation-induced changes in the abdomen. AB - After external-beam radiation therapy, radiation-induced changes may be observed in abdominal and pelvic organs at imaging. In the liver, an area of low attenuation corresponding to the radiation port (or an area of hyperattenuation if the underlying liver tissue shows fatty change) can be seen at computed tomography (CT) performed within 3-6 months after therapy. Later, the liver may be fibrotic and contracted. In the stomach, small intestine, and colon, wall thickening and edema are early manifestations. Ulcers may also be observed. Long term complications include strictures and fistulas. After irradiation of the kidneys, altered attenuation of the renal parenchyma may be seen at CT. Ureteral strictures, typically involving the distal ureter, may be observed after pelvic irradiation. The bladder may be small and contracted with a thickened wall after radiation exposure. Fistulas between the bladder and other pelvic organs sometimes occur. Typical musculoskeletal changes include growth abnormalities in skeletally immature patients, fatty replacement of bone marrow, and radiation osteitis. Radiation-induced neoplasms are also recognized after therapy. PMID- 9397459 TI - Hilar cholangiocarcinoma: thin-section spiral CT findings with cholangiographic correlation. AB - Hilar cholangiocarcinoma, a highly lethal tumor, is difficult to diagnose with conventional computed tomography (CT) because of its small size. Spiral CT allows more effective evaluation of these small lesions and better demonstrates the status of the hepatic arterial or portal venous circulation. Among 27 patients with hilar cholangiocarcinoma (infiltrative in 21, exophytic in two, polypoid in one, diffuse in three), thin-section spiral CT allowed identification of each tumor as an area of focal wall thickening that obliterated the lumen. Seventeen of the infiltrative tumors (81%) showed high attenuation. Identification of the level of biliary obstruction was possible in 63% of the patients (17 of 27). The level of obstruction was underestimated in six patients and overestimated in four. Spiral CT is a valuable method for diagnosis of hilar cholangiocarcinoma; however, spiral CT is less accurate in evaluation of intraductal tumor extent because of the limited z-axis resolution. PMID- 9397460 TI - Use of computed radiography in the study of an historic painting. AB - The authors demonstrate the use of radiography in the investigation of an historic painting and describe the potential benefits of computed radiography compared with conventional screen-film radiography. The subject for the comparison was a 16 x 19-foot oil-on-canvas painting, Scipio Africanus Freeing Massiva, by Giovanni Battista Tiepolo. Radiographs of the painting were obtained by using a portable, industrial radiographic unit and both conventional screen film and photostimulable phosphor plate cassettes. For this investigation, computed radiography had a number of advantages over screen-film radiography, largely due to its wider dynamic range and its capabilities for enhancing the digital images with image processing tools such as magnification, edge enhancement, colorization, and airbrushing. The ability to electronically combine images from the large painting into a single composite image file was extremely valuable, as this technique was much less cumbersome and resulted in much higher quality composite images than could be achieved with conventional radiography. An additional advantage of computed radiography includes the capability to easily archive and transmit these images in a digital format for subsequent review. PMID- 9397461 TI - Primary central nervous system lymphoma: radiologic-pathologic correlation. AB - Once an extremely rare neoplasm, primary lymphoma of the central nervous system (CNS) now ranks behind only meningiomas and low-grade astrocytomas in prevalence. Understanding of primary CNS lymphoma has increased greatly in recent years as a result of special immunohistochemical stains. Virtually all primary CNS lymphomas are composed of B cells. Although a viral cause has been suggested in some cases, the exact cause of the disease is still under investigation. Primary CNS lymphoma has a distinct affinity for perivascular extension. Although granular nodules may be seen at gross pathologic inspection, diffuse microscopic spread is always present, which accounts for the ability of this tumor to produce distant disease and local recurrences. The dense cellularity of the tumor and its predilection for the periventricular region also explains its typical hyperattenuated appearance on unenhanced computed tomographic scans and hypointensity on T2 weighted magnetic resonance images. Virtually all lesions enhance with contrast material. Although the overall prognosis for patients with primary CNS lymphoma remains poor, some advances have been made with radiation therapy and chemotherapy for this once uniformly fatal disease. PMID- 9397462 TI - The AAPM/RSNA physics tutorial for residents. X-ray generators. AB - The x-ray generator delivers the electrical power to energize the x-ray tube and permits the selection of x-ray energy, x-ray quantity, and exposure time. Major internal components of the generator include transformers, diodes and rectifier circuits, filament and stator circuits, timer switches, and kilovolt and milliampere meters. Single-phase, three-phase, high-frequency, and constant potential generators produce different voltage waveforms (ripple) and x-ray beam spectra. Phototimer and automatic brightness control subsystems measure radiation exposure incident on the image receptor to give instantaneous feedback for optimal radiographic film densities and fluoroscopic image brightness, respectively. At the generator control console, the operator sets the tube voltage, tube current, exposure time, phototimer film density, spot film acquisition, and fluoroscopic parameters. Selection of generator power and options depends on the intended clinical use. X-ray tube focal spot size and power loading capability should be matched to the x-ray generator and clinical imaging requirements. Single and multiple exposure rating charts as well as anode and housing thermal characteristic charts indicate power input and dissipation rates specific to a generator and x-ray tube target and housing. PMID- 9397463 TI - Value of high-frequency US for preoperative assessment of skin tumors. AB - The purpose of this study was to evaluate the accuracy of high-frequency ultrasound (US) in the preoperative assessment of skin tumors. A US scanner with a 20-MHz probe was used to visualize and evaluate 70 skin lesions (38 clinically suspected melanomas and 32 suspected basilar cell carcinomas [BCCs]) before surgical resection. A US morphologic study and a Doppler analysis of vascularity were performed for each tumor. Of the 70 tumors, 62 were clearly visualized, including 19 melanomas, 12 nonmalignant nevi, and 31 BCCs. Most lesions were hypoechoic. In 13 of 19 proved melanomas, the difference between the histologic and US measurements was equal to or less than 0.2 mm. Vessels were visualized in melanomas with thicknesses greater than 3 mm. All BCCs were visualized, and in 29% of cases of BCC, tumor size at US was greater than that at clinical examination. High-frequency, high-resolution US is a simple, reliable, noninvasive method for accurate preoperative assessment of skin tumor dimensions. This technique allows surgical planning to be adapted and reexcision to be avoided. However, its role is limited in the differential diagnosis of malignant and benign skin lesions. PMID- 9397464 TI - Improving the interactivity and functionality of Web-based radiology teaching files with the Java programming language. AB - Java is a programming language that runs on a "virtual machine" built into World Wide Web (WWW)-browsing programs on multiple hardware platforms. Web pages were developed with Java to enable Web-browsing programs to overlay transparent graphics and text on displayed images so that the user could control the display of labels and annotations on the images, a key feature not available with standard Web pages. This feature was extended to include the presentation of normal radiologic anatomy. Java programming was also used to make Web browsers compatible with the Digital Imaging and Communications in Medicine (DICOM) file format. By enhancing the functionality of Web pages, Java technology should provide greater incentive for using a Web-based approach in the development of radiology teaching material. PMID- 9397465 TI - Content preauthoring: preparing medical imaging information for multimedia authoring and quizzing. AB - Recent advances in multimedia development software and related hardware have given professionals and nonprofessionals tremendous power and flexibility to create multimedia education and training programs. Nevertheless, content organization remains a key and often neglected component of program development. Content preauthoring puts findings, diagnoses, differential diagnoses, and other standard radiologic concepts into a format that fosters logical program layout, centralized remediation, record keeping, decreased data entry, a variety of user levels, easy addition of cases, and linkage to a question-generating program. The goal of content preauthoring is to organize radiologic material into a hierarchical or spreadsheet-based structure that provides a logical basis for software design. By separating content design from software authoring, both processes become more manageable. This approach is applicable to visually oriented topics that focus on identification. The highly structured, goal oriented nature of the method makes it particularly suitable for newcomers to multimedia authoring. PMID- 9397466 TI - Intracranial neurenteric cysts: a differential diagnosis and review. PMID- 9397468 TI - US case of the day. Emphysematous pyelonephritis. PMID- 9397469 TI - Pediatric case of the day. Rubinstein-Taybi syndrome. PMID- 9397467 TI - General case of the day. Allergic (or hypersensitivity) bronchopulmonary aspergillosis (ABPA). PMID- 9397470 TI - Breast imaging case of the day. Invasive papillary carcinoma. PMID- 9397471 TI - Position, projection, method or view? PMID- 9397472 TI - Focal film distance in chest radiography. PMID- 9397473 TI - 3-D imaging: basic concepts for radiologic technologists. AB - This article describes the physical principles and clinical applications of three dimensional imaging in diagnostic radiology. It explores the history of 3-D imaging in medicine and reviews basic 3-D concepts. In addition, it discusses the technical aspects of medical 3-D imaging, including data sources, creation of 3-D space and rendering techniques. The article concludes with an overview of the clinical applications of 3-D imaging in computed tomography and magnetic resonance imaging, as well as a commentary on the future of 3-D imaging in radiology. PMID- 9397474 TI - The weighted abduction Grashey shoulder method. AB - The Grashey shoulder position is used to demonstrate damage to the glenohumeral joint caused by osteoarthritis, sclerosis, tumors, fractures, osteophytes and cystic changes. However, it can be difficult to assess loss of articular cartilage using the Grashey shoulder position because little axial load is applied to the glenohumeral joint. This article describes a method of creating a loading force across the glenohumeral joint by adding weighted arm abduction during the Grashey position to demonstrate loss of the articular cartilage. Case studies and radiographs are presented to discuss the advantages of using the Grashey position with weighted abduction. PMID- 9397475 TI - Gonadal protection methods in neonatal chest radiography. AB - The ideal method of gonadal shielding in the neonatal unit provides the greatest radiation protection while minimizing the potential for cross-infection. This study evaluated two common methods of gonadal shielding used during neonatal chest radiography--direct shielding and shadow shielding. For direct shielding, lead was placed over the gonadal region of a phantom. For shadow shielding, lead was placed on top of the crib. Results showed that direct shielding provided a greater reduction in gonadal dose than shadow shielding. PMID- 9397476 TI - 1997 Ed. C. Jerman Memorial Lecture. Charting our own course. PMID- 9397477 TI - A sneak preview of the sonography exam. PMID- 9397478 TI - Reflections on what makes a good teacher. PMID- 9397479 TI - Framed! PMID- 9397480 TI - Radiography of shoulder dislocations. PMID- 9397481 TI - Hydrocephalus: diagnosis and treatment. PMID- 9397482 TI - Need to justify a project? Write a proposal. PMID- 9397483 TI - Experts look at health care in 21st century. PMID- 9397485 TI - [Pathogenicity of Phylloporia chrysita (Aphyllophorales: Hymenochaetaceae) on Erythrochiton gymnanthus (Rutaceae)]. AB - The pathogenicity of Phylloporia chrysita (Berk.) Ryv. on Erythrochiton gymnanthus K. (Rutaceae) was studied in Carara Biological Reserve, seasonal Pacific of Costa Rica. Growth rate and distribution of basidiocarps were determined on health and diseased plants. P. chrysita caused 52% growth reduction on diseased plants. Fungal hyphae were observed on epidermis, parenchyma and vascular tissue, where they caused cellular breakdown. PMID- 9397486 TI - Intracellular calcium signalling in striated muscle cells. AB - Calcium signalling in cells is dependent on a communication between channels/ transporters in two membrane structures: the cell membrane and the membranes of endo- and sarcoplasmic reticula (ER/SR). In general, cytosolic Ca2+ can be raised by influx of calcium over the cell membrane through three types of channels: voltage-, receptor-, and store-operated channels (VOCs, ROCs and SOCs). This small Ca2+ influx is most often amplified by a Ca2+ release from the ER/SR through two types of channels: the IP3-receptor and the ryanodine receptor (RyR), which are huge proteins identified and cloned in recent years. We focus on the 'synaptic' connection between VOCs (L-type calcium channels) and RyRs of the SR in heart and skeletal muscle. Depolarization of the cell membrane (an action potential) opens the VOC and moves it in the membrane. One VOC triggers opening of a certain number of underlying RyRs that together release a quantum of calcium from the SR, a calcium spark. The communication between the VOC and RyRs is probably achieved primarily by a mechanical link in skeletal muscle (voltage controlled calcium release), and by the small inward calcium flux through the VOC in the heart (calcium-induced calcium release, CICR). Conditions as different as heart failure, myasthenia gravis, malignant hyperthermia, and skeletal muscle fatigue, may be examples of deteriorated control or function of the RyR. PMID- 9397487 TI - Ryanodine binding sites measured in small skeletal muscle biopsies. AB - A method allowing measurement of the concentration of [3H]ryanodine binding sites in small skeletal muscle specimens (> 10-20 mg) was developed. A membrane fraction containing 87% of the [3H]ryanodine binding sites of the tissue and exhibiting one single KD of 18-27 nmol l-1 in rat and 8 nmol l-1 in human muscles (p < 0.05) was obtained. Maximum binding to rat EDL and soleus muscles equalled 59.1 and 16.2 pmol g-1 wet wt, whereas in human gluteus muscles binding was 12.3 pmol g-1 wet wt. The [3H]ryanodine binding showed a dependency on Mg2+ and pH similar to previously published results. As measured by Ca2+ selective mini electrodes, the [Ca2+] causing 50% of maximum [3H]ryanodine binding (K0.5) was 200-400 nmol l-1 for different muscles. [Ca2+] higher than 1 mmol l-1 caused strong inhibition of the [3H]ryanodine binding, and both high and low [Ca2+] caused rapid dissociation of the complex. At ionic strength lower than 100 mmol l 1, more than 50% of the [3H]ryanodine was bound to particles with size less than 1.2 microns which were not retained by GF/C filters. Thus, we have obtained an almost complete quantitative recovery of functional RyRs from small muscle specimens exhibiting high affinity for Ca2+, which stimulated ligand binding. PMID- 9397488 TI - Pamidronate and biochemical markers of bone turnover. AB - We have examined the response of different biochemical bone turnover markers to intravenous pamidronate administration (15 mg for 5 days) in 14 patients with Paget's disease, on days 8, 15 and 30 after pamidronate treatment. Urinary parameters of bone resorption, free pyridinolines (Pyr) and hydroxyproline (OHP), as well as serum tartrate-resistant acid phosphatase (TRAP) were measured. Two serum biochemical osteoblastic markers, alkaline phosphatase (AP) and osteocalcin (OC), were also analysed. In addition, ionic calcium (Ca2+) was measured in blood, and parathyroid hormone and calcitriol were measured in serum. All the biochemical markers of bone resorption tested decreased throughout the study. TRAP levels decreased slowly, meanwhile Pyr decreased maximally, below OHP values on day 8. However, the latter were lowest and were lower than those of Pyr on days 15 and 30. AP serum values also decreased during the study. Conversely, OC serum levels increased on days 8 and 15, decreasing to baseline levels on day 30. Ca2+ blood levels decreased while PTH plasma levels increased at all times during the period studied. Calcitriol serum levels increased on day 15. In conclusion, intravenous pamidronate administration was found to modify several biochemical parameters of bone turnover, including Pyr. Moreover, the changes in these parameters were different in intensity and "time course" during the study. PMID- 9397489 TI - Effects of adrenergic and muscarinic agonist stimulation on IP3 and cyclic nucleotide levels in the pressure overloaded rat heart. AB - In this study, the dynamic interrelationships between myocardial functional state and changes in the second messenger content in pressure-overloaded hypertrophied hearts were investigated. Forty-three rat hearts were used after partial clamping of the abdominal aorta. The isolated hearts were perfused with Krebs-Henseleit buffer and allocated to perfusion for 20 s or 40 min as controls (n = 12); or with noradrenaline (10(-6) mol l-1, n = 11); carbachol (3 x 10(-7) mol l-1, n = 9); or noradrenaline plus carbachol (10(-6) mol l-1 + 3 x 10(-7) mol l-1, respectively, n = 11). maxdP/dt increased more than 2-fold already after 20 s on noradrenaline stimulation, followed by a significant increase in cAMP. After 40 min, maxdP/dt was lower than the maximal value, although higher than controls. cAMP was also decreased, but still significantly higher than controls. Perfusion with noradrenaline plus carbachol produced the same changes in maxdP/dt as those seen after noradrenaline stimulation alone, but failed to increase cAMP content after both 20 s and 40 min. The inositol trisphosphate (IP3) content was increased 40 min of control perfusion (p < 0.05). Noradrenaline and carbachol, separately, produced an increase in IP3 content already after 20 s (p < 0.05). The combination of noradrenaline plus carbachol also produced an increase of IP3 (p < 0.05; compared to controls), but to a lesser extent when compared either to noradrenaline or carbachol (p < 0.05). After 40 min of perfusion, IP3 was in the same range regardless of added agonist(s) and still slightly above control level (p < 0.05). The early increase in maxdP/dt induced by noradrenaline or the combination of noradrenaline plus carbachol was not paralleled by a decrease in ATP content. This was also the case upon addition of carbachol alone. However, after 40 min of agonistic perfusion, ATP levels were substantially decreased. In conclusion, myocardial IP3 content in pressure-overloaded hypertrophied hearts was not different from that of sham-operated hearts. After agonistic stimulation, an early increase in IP3 formation was seen. Attenuation of the IP3 response by combined stimulation with noradrenaline and carbachol was initially present in pressure-overloaded hypertrophied hearts. After 40 min no attenuation was found for either IP3 or for cAMP content, suggestive of induction of a desensitization. PMID- 9397490 TI - Poor metabolic control, early age at onset, and marginal folate deficiency are associated with increasing levels of plasma homocysteine in insulin-dependent diabetes mellitus. A five-year follow-up study. AB - In a previous study, we showed that diabetic patients exhibited significantly increased concentrations of total plasma homocysteine (tHcy), but not until the onset of nephropathy. It was suggested that the hyperhomocysteinaemia might contribute to the accelerated atherosclerotic process in diabetic patients. In the present study, we have analysed the main determinants of plasma homocysteine (i.e. serum cobalamin, blood folate and serum creatinine), and also some other parameters related to diabetes mellitus, such as medical history, metabolic and renal quantities, on two occasions with a 5-year interval in 50 patients with insulin-dependent diabetes mellitus, in order to further elucidate the relation between plasma tHcy and diabetes mellitus. The result of the present study shows that diabetic patients with the lowest age at onset and with the poorest metabolic control are those most prone to a rapid increase in plasma tHcy concentration. The increment in plasma tHcy concentration in this group of patients may at least partly be explained by a marginal deficiency of blood folate concentrations. PMID- 9397491 TI - Detection of tumour DNA in serum of colorectal cancer patients. AB - Circulating tumour DNA has previously been detected in serum and plasma of patients with lung cancer and head and neck cancer. These observations could potentially lead to new, specific and non-invasive tools for diagnosis, prognosis and follow-up in neoplastic disease, if found to be a more general phenomenon. To test if tumour DNA is also present in serum of patients with colorectal cancer, we selected 14 colorectal cancer patients with advanced disease. In seven patients, K-ras mutations were detected in the primary tumour, using mutant specific primers for point mutations in codon 12 or 13 of the K-ras gene. All patients were analysed for mutant DNA in serum. Tumour-specific point mutations, corresponding to the K-ras mutations found in the primary tumour were detected in the serum of all patients but one. No mutant K-ras could be detected in the serum of seven patients without K-ras mutations in the primary tumour. These results may be useful in assessing tumour burden in patients with neoplastic disease. Moreover, consecutive testing of serum tumour DNA after surgery or chemotherapy may be used as a tumour marker for recurrent disease. PMID- 9397492 TI - Multicentre evaluation of the urine analyser Miditron Junior. AB - A multicentre evaluation of the urine analyser Miditron Junior was performed at four laboratories. The Miditron Junior analyser provides semi-quantitative results for erythrocytes, bilirubin, urobilinogen, ketone bodies, glucose, protein, nitrite, leukocytes, pH and relative density. Accuracy of the Miditron Junior analyser was evaluated by comparison to quantitative analytical chemistry methods (glucose, total protein), physical methods (pH, relative density), and microscopic methods (erythrocytes, leukocytes). Agreement was defined as identical or neighbouring concentration block. The level of agreement found with chemical, physical or microscopic methods for the six analytes tested varied from 79 to 99%. The within-run precision was determined as repeatability by using 31 native urines. The results of repeated measurements (n = 10) fell in the same concentration block, or in case of borderline concentration were spaced between two adjacent colour blocks. Day-to-day precision covering a minimum of 20 days using commercially available control solutions yielded results within +/- one colour block from the mean. PMID- 9397493 TI - An external quality assessment study on the analysis of methylmalonic acid and total homocysteine in plasma. AB - In spite of the increasing interest in the analysis of methylmalonic acid and total homocysteine in plasma, data on interlaboratory variation is lacking. We report the results of an external quality assessment study with the participation of 15 laboratories in the Scandinavian countries performing these analyses on a regular basis. For methylmalonic acid, using serum, heparin fluoride plasma and an aqueous sample, CVs were found in the range of 11-17%. For total homocysteine using EDTA plasma, heparin fluoride plasma and an aqueous sample, CVs were in the range of 6-12%. For both analytes, a significant correlation between the individual recoveries of added analyte and the results for the aqueous sample was found, suggesting that the use of inconsistent calibrations in the participating laboratories are contributing to the interlaboratory variation. An acceptable range for results from the individual laboratory was calculated using data on the biological within-subject and between-subject variations reported in the literature. These ranges were violated by several laboratories when using the consensus mean or median as target values. Even if the results of the present study document a reasonable standard in the measurement of methylmalonic acid and total homocysteine in plasma in the participating laboratories there is room for improvement and a permanent scheme of external quality assessment using relevant samples is essential. From 1997, a regular scheme has been available from our laboratory. PMID- 9397494 TI - 99mTc-labelled immunoglobulin scintigraphy in arthritis: an analysis of synovial fluid activity. AB - The distribution of 99mTc-labelled human polyclonal non-specific immunoglobulin G (HIG) in the synovial fluid was studied in 14 patients with rheumatoid and non rheumatoid arthritides. Analysis included the determination of the total activity per ml synovial fluid 6 h post-injection (p.i.) of the tracer as well as of the protein- and cell-bound fractions. At 6 h p.i., > 60% of the injected dose remained in plasma as protein-bound radioactivity. Values in the synovial fluid ranged between 0.001 and 0.009% of the injected dose per ml. Importantly, the synovial fluid to plasma ratio was consistently < 1 (range: 0.09-0.43), which is in the range of ratios observed for endogenous proteins in vivo. Similar values were obtained in samples of synovial tissue obtained at surgery in two patients. These data are consistent with the hypothesis that labelled HIG accumulates in the extracellular fluid (both within the synovial tissue and fluid) by non specific mechanisms (such as increased blood pool and capillary permeability) and does not equilibrate with circulating plasma proteins in accordance with basic knowledge of synovial physiology. In addition, it was found that most of the activity remained bound to the proteins in the fluid and that cell-binding occurred to a very low degree that cannot be considered an important mechanism of uptake of this radiolabelled agent in vivo. These results provide the first evidence in an in vivo human setting that radiolabelled HIG accumulates mainly by non-specific mechanisms in inflamed joints. PMID- 9397495 TI - Screening for EDTA-dependent deviations in platelet counts and abnormalities in platelet distribution histograms in pseudothrombocytopenia. AB - Screening for pseudothrombocytopenia caused by in vitro platelet clumping has been performed in 45,000 subjects attending a general hospital. In our region, the observed prevalence of EDTA-induced pseudothrombocytopenia in blood samples with an initial platelet count below 150 x 10(9)/l was estimated to amount to 0.1%. EDTA-induced pseudothrombocytopenia was confirmed by detection of platelet aggregates by means of microscopic evaluation from the blood smear. In routine investigations, pseudothrombocytopenia could be highly suspected when the Sysmex NE 8000 showed characteristic peculiarities in the white blood cell (WBC) scattergram and histogram. Platelet aggregation is avoided in such cases by the use of citrate as an anticoagulant instead of EDTA. Pseudothrombocytopenia was detected in 46 subjects. As a screening test for pseudothrombocytopenia, increased cut-off values derived from the WBC histogram demonstrated 90% sensitivity and 100% specificity. Automated flagging for platelet clumps, deviations reflecting MPV, or PDW abnormalities revealed lower scores with respect to sensitivity. PMID- 9397496 TI - Preconditioning the globally ischaemic, isolated rat heart: the impact of the preconditioning model on post-ischaemic systolic and diastolic function. AB - In studies of preconditioning, a variety of models have been used. The aim of the present study was to find the optimal preconditioning model for preservation of cardiac function during reperfusion of globally ischaemic, Langendorff-perfused rat hearts. Cardiac function was assessed by the occurrence of severe reperfusion arrhythmias (ventricular fibrillation or asystolia), heart rate (HR), left ventricular systolic (LVSP), end diastolic (LVEDP), and developed pressures (LVDP = LVSP - LVEDP), as well as coronary flow (CF). Series 1 (n = 17) in each group: control perfusion for 20 min without preconditioning or 2 episodes of 2, 3, 4, or 5 min of ischaemia, each followed by 5 min reperfusion, before 25 min ischaemia and 60 min reperfusion. Preconditioning reduced the incidence of reperfusion arrhythmias, attenuated the reperfusion-induced increase of LVEDP, and increased CF, but did not influence LVSP, LVDP, or rate x pressure-product (RPP = LVSP x HR) during reperfusion. The greatest effect was found by 2 min ischaemia and 5 min reperfusion. In series 2 (n = 17 in each group) control perfusion for 7 or 28 min, or preconditioning with 1-4 episodes of 2 min ischaemia and 5 min reperfusion before 35 min ischaemia and 60 min reperfusion were compared. Reduction of severe reperfusion arrhythmias and LVEDP elevation, as well as improvement of CF, LVDP, and HR in preconditioned hearts were observed in series 2. Optimal cardioprotection was achieved by only one episode of preconditioning. In conclusion, preconditioning before global ischaemia improved cardiac function during reperfusion of isolated rat hearts. The most marked effects were reduction of severe reperfusion arrhythmias and attenuation of diastolic dysfunction. Although all preconditioning models employed were cardioprotective, 1 episode of 2 min ischaemia provided optimal protection. PMID- 9397497 TI - Use and usefulness of laboratory handbooks. AB - The aim of this study was to investigate how reference handbooks distributed by hospital laboratories are used by medical doctors, and to what extent this kind of information can influence or change doctors' work habits. We also wanted to see if books with various contents of information are valued differently by the users, and we asked for preferences for an ideal book. A questionnaire was sent to 2075 medical doctors served by five Norwegian hospital laboratories. The overall response rate was 66%, of whom 76% had received a handbook. Seventy-eight percent of respondents who stated that they had received a handbook kept it in their consulting room and 45% used it once or more weekly. The majority (89%) found the books beneficial in their everyday work. Many doctors (36%) claimed that they had changed their routines as a result of the information in the book. The way of interpreting test results was influenced most often, followed by indications for ordering laboratory tests, sample collection and specimen handling, and patient preparation. Nearly all respondents (97%) felt that handbooks of this kind are beneficial to their technical and nursing staff. The results show that handbooks distributed by medical laboratories are well received, frequently used and highly appreciated by medical doctors. Comprehensive books are rated higher than smaller books. PMID- 9397498 TI - Rectal administration of N-acetylcysteine in swine: a pilot study. AB - The purpose of this pilot study was to determine if N-acetylcysteine (NAC) administered via the rectal route in swine is absorbed into the systemic circulation. Fasting swine were anesthetized, intubated, monitored and i.v. access was obtained by femoral cutdown. NAC was administered into the rectal vault (2.0 g/kg) via a balloon-tipped Foley catheter inserted into the animals' rectum. NAC administered via the rectal route resulted in systemic absorption as determined by spectrophotometric methods in 5 of the 7 study animals. This study provides important information regarding the development of a potential alternative route for the administration of NAC. PMID- 9397499 TI - Toxicity of waterproofing spray is influenced by the mist particle size. AB - In a previous study, we showed that waterproofing sprays that are toxic generate mists with smaller particles than do nontoxic products. In this study, we made 4 waterproofing sprays (A, B, C and D) with identical ingredients but with different mist particle sizes and compared the pathological changes produced in the lungs of mice. The mist particle diameters were 32.8 +/- 3.2, 62.0 +/- 3.8, 89.1 +/- 4.1 and 143.2 +/- 5.0 microns for sprays A, B, C and D, respectively. Pathological lung changes were evaluated by a 6-criteria grading system (thickening of the alveolar septum, cellular infiltrations in the alveolar septum, alteration of the bronchial mucous membrane, hyperemia of the alveolar wall, transudative hemorrhage, and alveolar collapse). Sprays A and B caused significantly greater scores as compared to the control group for all the criteria except mucosal changes, whereas the changes from sprays C and D were slight and the differences in scores were not significant. These results suggest that toxicity of waterproofing spray is influenced by the mist particle size generated and may help manufacture safer waterproofing spray products. PMID- 9397500 TI - Dietary fumonisins disrupt sphingolipid metabolism in mink and increase the free sphinganine to sphingosine ratio in urine but not in hair. AB - This study was conducted to investigate the effects of dietary Fusarium moniliforme culture material (M-1325) containing known concentrations of fumonisins B1, B2 and B3 on sphingolipids in urine and hair of mink (Mustela vison) for use as potential, non-invasive biomarkers of exposure to fumonisins in this species. Feeding mink diets containing 86, 22, and 7 ppm or 200, 42, and 12 ppm of fumonisins B1, B2 and B3, respectively, yielded marked increases in urinary free sphinganine (Sa) and free sphingosine (So) concentrations, and free Sa/free So ratios (2 to 11-fold) within 7 d, compared to controls. Free Sa and free So concentrations and Sa/So ratios in hair samples from mink fed the control or high dose fumonisin diets for 100 days were similar and were not apparently altered by exposure to these mycotoxins. These results suggest that Sa/So ratios in urine, but not in hair of mink can serve as an early indicator of exposure to fumonisins in this species. PMID- 9397501 TI - A modified electrometric method for measurement of erythrocyte acetylcholinesterase activity in sheep. AB - A modified method was compared with an original electrometric method for measurement of erythrocyte acetylcholinesterase (EChE) activity in sheep. The mean +/- SD (pH/30 min) of EChE activity of 8 sheep measured by the modified procedure (0.70 +/- 0.15) was not significantly different from that of the original method (0.64 +/- 0.12). The inherently low plasma cholinesterase activity of the sheep as measured by the 2 methods were also not significantly different from each other (0.09 +/- 0.04 vs 0.10 +/- 0.04). The coefficient of variation of the modified method in measuring EChE activity was 8%. The method was used to demonstrate in vitro inhibition of sheep EChE activity by the organophosphorus and carbamate insecticides dichlorvos and methomyl, respectively. The method could be well-suited for rapid measurement of EChE activity in sheep, especially in cases of organophosphate and possibly carbamate poisoning. PMID- 9397502 TI - Influence of alkalinization of glutaraldehyde biocidal solutions on acute toxicity, primary irritancy, and skin sensitization. AB - Aqueous glutaraldehyde (GA) is used at a concentration around 2% for the cold sterilization of endoscopy and dental instruments. Stock GA solution (pH 3.1-4.5) is alkalinized (pH 7.8-8.0) before use to optimize biocidal activity. The possible differential handling hazards between acidic unbuffered GA (UGA) and alkaline buffered GA (BGA) were compared for acute toxicity, primary irritancy and skin sensitizing potential using a 2.2% GA solution. Peroral LD5.0 values (with 95% confidence limits) in rats (combined sexes) were 3.45 (3.13-3.80) g/kg for UGA and 4.16 (3.13-5.52) g/kg for BGA; signs and gross pathology were similar. A 24-h occluded cutaneous application of 16.0 g/kg in the rabbit did not produce mortality; moderate skin irritancy was observed. No systemic effects occurred with UGA and only a few with BGA (unsteady gait, sluggishness, rapid breathing). Local skin irritation from a 4-h occluded contact with 0.5 ml was relatively minor and slightly more marked with BGA than UGA. Rats exposed to a statistically generated saturated vapor atmosphere for 6 h did not show any signs or gross pathology, and only slight weight loss occurred (UGA females). Rabbit eye irritation studies (0.1 ml) showed slightly more marked conjunctival reactions with BGA, but corneal injury was marked and persistent with BGA and only slight and transient with UGA. With 0.01 ml, no corneal injury occurred, but conjunctival reaction was more marked with UGA. A guinea pig maximization study showed UGA to produce a higher sensitizing index (68% at challenge, 32% at rechallenge) than BGA (30% at challenge, 5% at rechallenge). Severity indices at challenge was also higher for UGA [0.84 (24 h), 0.47 (48 h)] than BGA [0.45 (24 h), 0.18 (48 h)]. Both UGA and BGA have generally similar acute toxicity and skin irritancy; BGA has greater corneal injuring potential, and UGA has a greater skin sensitizing potential. PMID- 9397503 TI - New treatment regimens in organophosphate (diazinon) and carbamate (methomyl) insecticide-induced toxicosis in fowl. AB - The objective of this work was to determine optimal treatment regimens for organophosphate (OP) or carbamate insecticide toxicoses in fowl using the antidotes atropine sulfate and pralidoxime chloride (2-PAM). Broiler chicks in treatment groups, each comprising 3 replicates of 6-7 birds/replicate, were gavaged on a body weight (BW) basis with the OP and carbamate insecticides, diazinon and methomyl, respectively, at lethal dosages. Treatment groups were injected with either or both of the antidotes at various dosages as soon as clinical signs appeared. Birds appearing healthy 24 h thereafter were regarded as having been treated successfully. At a dosage of 100 mg/kg BW, atropine was mildly toxic and at 200 mg/kg 2-PAM was severely toxic (but not lethal), whereas at dosages of 50 and 100 mg/kg BW, respectively, the antidotes were at their most effective. With diazinon, atropine alone was only partially effective (12/20 survivors), whereas 2-PAM was extremely efficacious. (20/20 survivors); the combination of the 2 antidotes at 2 dosages was slightly less effective (19/20 survivors) than 2-PAM alone. For methomyl toxicity, atropine was largely successful (18/20 survivors), whereas 2-PAM was mostly unsuccessful (10/20 survivors); the combination at high dosage was less effective (15/20 survivors) than atropine alone, but at a low dosage the combination was the most successful (20/20 survivors). The results indicate that anticholinesterase insecticide toxicoses in fowl should not be treated according to textbook recommendations, and antidotal dosage with atropine should be up to 100 times greater than is commonly recommended. The specific cause of the toxicoses should ideally be determined before treatment is given, but as this is often unknown, a combination of antidotes may be the optimal treatment protocol. PMID- 9397504 TI - Marijuana (Cannabis sativa) toxicosis in cattle. PMID- 9397505 TI - Flurazepam toxicosis in two dogs. AB - Flurazepam is a benzodiazepine (BZD) derivative and a category IV controlled substance. It is a widely prescribed hypnotic drug for use in sleep disorders. Two dogs were maliciously poisoned with this drug and died. Flurazepam was detected in the urine of 1 dog by thin-layer chromatography. Flumazenil, an antagonist for BZD receptors, is currently used in humans to reverse the effects of intoxication with BZD. It may also be of use in treating companion animals. PMID- 9397506 TI - Neurotoxicity and secondary metabolic problems associated with low to moderate levels of exposure to excess dietary sulphur in ruminants: a review. AB - Problems associated with a low to moderate excess in dietary sulphur (S) intake in ruminants are being increasingly recognized. Comparing more recent reports with older data, there is an evident decrease in tolerance of cattle and sheep for even moderately elevated levels of dietary S, and an apparent drastic change in the clinical picture of chronic dietary S toxicoses. Outbreaks of polioencephalomalacia (PEM) in ruminants in association with excess dietary S have been reported in recent years throughout the world. Excessive levels of S containing compounds in domestic ruminant animals' rations, and clinical problems associated with low to moderate levels of exposure to dietary S may be more common than previously thought. This review presents a comprehensive evaluation of the problems associated with excessive levels of S in ruminants' rations. Emphasis is placed on the recently increasing incidence of S-induced PEM. Secondary metabolic disorders associated with excessive intake of S are also discussed. PMID- 9397507 TI - Poisonings in animals: the 1993-1994 report of the American Association of Poison Control Centers. AB - More than 82% of the 140,614 animal poison exposures reported in 1993 and 1994 occurred in dogs and almost 14% occurred in cats. Almost all reported were acute exposures to a single product. Tables of detailed data are provided. PMID- 9397508 TI - Instant methods to spot-check poisonous podophyllum root in herb samples of clematis root. PMID- 9397509 TI - Antihistamine-containing cough/cold medications present a low hazard in pediatric accidental exposure incidents: analysis of Poison Control Center data. AB - We studied accidental exposure to pediatric cough/cold medications in children under 6-y-of-age to determine whether the presence of an antihistamine (chlorpheniramine) in the product increased the likelihood for adverse outcomes. General accidental exposure cases reported to the American Association of Poison Control Centers (AAPCC) during 1988-1992 were analyzed for specific over-the counter cough/cold pediatric products containing identical concentrations of active ingredients except for the presence or absence of chlorpheniramine. These case reports were evaluated for differences in medical outcome, symptom assessment, management site and therapy, as coded by poison control centers participating in the AAPCC Toxic Exposure Surveillance System. A total of 10,289 cases of accidental exposures were evaluated for the specific products included in this analysis. While these cases represented a small percentage of total exposures to these products (approximately 3% of total cases in children under 6 y-of-age reported to the AAPCC for all cough/cold medications during 1988-1992), they provided a unique opportunity to evaluate the impact of the antihistamine component of the formulation. This study demonstrated that the presence of chlorpheniramine did not affect the medical outcome or the extent to which symptoms were reported. Additionally, similar percentages of exposures for these 2 products were managed at the site of the incident, and required either no therapy or only required fluids and observation. There were no notable differences in the percentage of cases which involved more aggressive treatment procedures (activated charcoal, cathartic, lavage). This analysis demonstrates that over-the-counter cough/cold medications containing chlorpheniramine present low potential for hazard in cases of accidental ingestions in young children and do not show an increased likelihood for adverse outcomes of accidental exposures compared to cough/cold medications not containing this antihistamine. PMID- 9397510 TI - Trends in hospitalizations and mortality due to medicinal or non-medicinal poisonings in Hong Kong. AB - Previous epidemiologic studies of poisonings in Hong Kong are regional hospital or poison information center-based and have focused on either adults or children. This paper reports on the territory-wide hospitalization and mortality rates, comparing medicinal and non-medicinal poisonings in the general population. Between 1980 and 1995, the figures for hospitalizations and mortality due to medicinal (ICD codes 960-977) or non-medicinal (ICD codes 980-989) poisonings were obtained from the Annual Reports of the Department of Health, Hong Kong Government. Rates of medicinal poisonings increased between 1980/81 (57.3/100,000) and 1987/88 (80.9/100,000), but then declined (59.1/100,000) in 1993/94. Rates of non-medicinal poisonings were rather static (49-53/100,000) between 1980/81 and 1988/89, but then declined (22.0/100,000) in 1994/95. Between 1980/81 and 1988/89, rates of fatal medicinal poisonings (0.73-1.31/100,000) were similar to those of fatal non-medicinal poisonings (0.98-1.70/100,000). However, from 1989/90, there was an increase in the rates of fatal medicinal poisonings (1.94-2.80/100,000), although rates for non-medicinal poisonings remained much the same (0.80-1.38/100,000). Hospitalizations due to poisonings are now less common in Hong Kong than before, due largely to a greater decline in non medicinal poisonings. PMID- 9397511 TI - Catnip and the alteration of human consciousness. AB - Uncertainty exists regarding the ability of catnip (Nepeta cataria) to affect human consciousness. We report a case of a toddler exhibiting central nervous system depression after consuming a large quantity of catnip. His obtundation was not attributable to another cause. We review the published literature describing the alleged psychoactive capabilities of catnip and present our case as further information for use in this ongoing controversy. PMID- 9397512 TI - Photosensitization and crystal-association cholangiohepatopathy in cattle grazing Brachiaria decumbens in Brazil. PMID- 9397513 TI - Structural factors contributing to insecticidal and selective actions of neonicotinoids. AB - Nicotinoids and neonicotinoids are characterized by the presence of the 3 pyridylmethylamine moiety in their structure. In the former, the amino nitrogen atom is ionized, while in the latter the corresponding nitrogen atom is not ionized but bears a partial positive charge. Both types of insecticides interact with nicotinic acetylcholine receptor (nAChR) of insect origin. The poor interaction of neonicotinoids with vertebrate nAChR was shown by its poor binding affinity to the nAChR from Torpedo electric organ and rat brain and poor activation with nAChR expressed in Xenopus oocytes. The full positive charge was essential to interact with the vertebrate nAChR, while the 3-pyridylmethylamine moiety with a partial positive charge was enough to interact with the insect nAChR. For penetration into the insect central nervous system, hydrophobicity seemed to play an important role, as indicated by the binding of the injected compounds to the housefly head nAChR. The ionization reduced hydrophobicity and limited the penetration of nicotinoids, resulting in less insecticidal activity. Among neonicotinoids, nitromethylene type compounds, though far higher in binding affinity, were less hydrophobic than the corresponding nitroimine type, and the net result was better or inferior insecticidal activity. A chlorine atom at the 6 position of the 3-pyridyl group found in commercialized neonicotinoids contributes to increased binding affinity and more importantly hydrophobicity, thus increasing insecticidal activity. N-Me-imidacloprid was found to be a propesticide of imidacloprid. PMID- 9397514 TI - CYP6D1 protects thoracic ganglia of houseflies from the neurotoxic insecticide cypermethrin. AB - CYP6D1 is a housefly cytochrome P450 known to metabolize neurotoxic pyrethroid insecticides. To determine if the nervous system was capable of metabolizing pyrethroids, we examined CYP6D1-mediated in vitro metabolism in thoracic ganglia from pyrethroid-resistant (LPR) and -susceptible (CS) strains of housefly. SDS PAGE/immunoblotting revealed that CYP6D1 was expressed in all tagmata and in thoracic ganglia of both strains, but in all cases the levels of CYP6D1 were higher in the LPR strain. Using a CYP6D1-specific antiserum, we found CYP6D1 to be the major, and possibly the only, P450 isozyme involved in cypermethrin metabolism in thoracic ganglia homogenates. Additionally, thoracic ganglia homogenates from LPR houseflies metabolize more cypermethrin than preparations from susceptible flies. This metabolism was inhibited by piperonyl butoxide and a CYP6D1-specific antibody. Our results indicate that thoracic ganglia of LPR houseflies are protected from the neurotoxin cypermethrin by virtue of the higher levels of CYP6D1 compared to the susceptible houseflies. This P450-mediated detoxification of an insecticide at the level of the target tissue helps to explain the high levels of resistance to pyrethroids in the LPR strain. PMID- 9397515 TI - Effects of exposure to cypermethrin on saxitoxin binding in susceptible and pyrethroid-resistant houseflies. AB - Saxitoxin (STX) binding was measured in susceptible (SBO) and pyrethroid resistant (KDR) female houseflies having only target site insensitivity as a resistance mechanism. In KDR flies, there was a quantitative decrease in STX binding capacity (Bmax) relative to SBO flies coupled with an increase in binding affinity (Kd). Treatment of SBO flies with sublethal doses of cypermethrin resulted in a large decrease in the number of STX binding sites and an increase in STX binding affinity. In KDR flies, identical treatments had the opposite effects. Treatment of both strains with higher doses of cypermethrin resulted in smaller decreases in Bmax values coupled with decreases in binding affinities. The results show that physiological changes in STX binding occur upon exposure to extremely low doses of cypermethrin. The data suggest that the kdr resistant gene may be expressed as changes in STX binding kinetics and that measurements of STX binding in pyrethroid-treated insects may be a useful approach for studying pyrethroid's mode of action and resistance. PMID- 9397516 TI - Isolation of the V-ATPase A and c subunit cDNAs from mosquito midgut and Malpighian tubules. AB - Using conserved amino acid sequences for the design of oligonucleotide primers, we isolated cDNA clones for two subunits of the V-ATPase from the midgut and Malpighian tubules of Aedes aegypti larvae. The 3.1 kb cDNA of the A subunit of the peripheral catalytic V1 sector codes for a protein of 68.6 kDa. The protein contains conserved motifs, including an ATP/GTP binding site, found in all other A subunits. Southern analysis using the A subunit as a probe suggests the presence of only a single copy of gene in the Aedes aegypti. The 0.85 kb cDNA of the c subunit of the membrane H+ conducting V0 sector codes for a protein of kDa. This protein has four transmembrane domains and contains a conserved glutamic acid that serves as the binding site for dicyclohexylcarbodiimide. Southern analysis using the c subunit as a probe suggests the presence of more than one related gene in the genome of Aedes aegypti. Pileup analysis of various A and c subunits shows that these subunits fall into distinct clusters, including one in which all arthropod proteins are clustered. PMID- 9397517 TI - Two-photon excitation of ethidium bromide labeled DNA. AB - We examined the steady state and time-resolved emission of DNA stained with ethidium bromide (EB) when excited with 90 fs pulses from a mode-locked titanium sapphire laser. Over the wavelength range from 840 to 880 nm EB-DNA was found to display two-photon excitation, with a cross-section near 7 x 10(-50) cm4s/photon. Frequency-domain intensity decay measurements revealed similar multi-exponential intensity decays for one- and two-photon excitation. Time-resolved anisotropy decay measurements revealed similar correlation times, but different amplitudes as has been observed previously for two- versus one-photon excitation. These results indicate that two-photon excitation of EB-DNA can be accomplished with the fundamental output of a Ti:sapphire laser without obvious heating or perturbation of the DNA. PMID- 9397518 TI - A study of the dielectric properties of E. coli ribosomal RNA and proteins in solution. AB - The permittivity of ribosomal proteins and ribosomal RNA (rRNA) in solution was measured in the range 100 kHz to 1 GHz at four different temperatures (5, 15, 25 and 35 degrees C). The experimental dielectric relaxation was analysed by the Cole-Cole equation and, from the best-fit parameters, the average values of the dipole moment and molecular radius of the proteins were obtained. The activation enthalpy was calculated from an Arrhenius plot of the relaxation time. The energy involved in the dielectric polarization of free proteins has a magnitude of about one hydrogen bond. The data on RNA were analysed according to the Mandel model. This analysis allowed the calculation of the "subunit b" as defined by Mandel. This parameter is dependent on the temperature and therefore the relaxation time does not follow the Arrhenius law. Our data thus show that, in solution, the rRNA structure is thermally rather unstable and highly flexible. PMID- 9397519 TI - Interaction of DAPI with pepsin as a function of pH and ionic strength. AB - The fluorescent probe 4',6-diamidino-2-phenylindole (DAPI), extensively used with nucleic acids, has also recently been used with membranes and proteins (Favilla et al., Biophys. Chem., 46 (1993) 217-226 and Mazzini et al., Biophys. Chem. 52 (1994) 145-156, respectively). The spectroscopic changes of DAPI observed, namely a considerable enhancement of fluorescence, induced circular dichroism (CD) and absorbance spectral shifts, have been exploited to study the binding of this dye to both native and alkali denatured pepsin. Fluorescence and CD titrations show that nearly two molecules of DAPI bind to either native or alkali denatured pepsin with pH and ionic strength dependent Kd values, whereas absorbance titrations evidentiate an interaction characterized by a lower affinity and a larger number of binding sites. Therefore two kinds of interaction are proposed: a specific one, involving both hydrophobic and electrostatic forces; and a non specific one, involving surface protein negative charges only. Finally, the behaviour of DAPI as a competitive inhibitor and the remarkable effect of pepstatin A, a specific inhibitor of pepsin, on both the CD and fluorescence spectra of DAPI in the presence of pepsin, show the involvement of the protein active site in the interaction. PMID- 9397520 TI - Alkaline denaturation and partial refolding of pepsin investigated with DAPI as an extrinsic probe. AB - The binding parameters of DAPI to porcine stomach pepsin have been described in the previous article in this issue (A. Mazzini et al.). Here we exploit the differences in the spectroscopic (fluorescence and circular dichroism) properties of DAPI bound to either native or alkali denatured pepsin. We follow the kinetics of pepsin denaturation around neutrality (pH range 6.8-7.4), at several phosphate buffer ionic strengths (range 0.02-0.25). The dependence of the apparent dissociation rate constant on pH clearly shows that the rate limiting step follows the dissociation of about three acidic protein residues. The accelerating effect by ionic strength we observed can be accounted for by a simple treatment based on both transition state theory and Debye-Hueckel's limiting law. Furthermore, when a solution of pepsin, rapidly denatured at pH 7, is reacidified to a pH between 4.5 and 5.5, a substantial recovery of protein secondary structure, with no enzymatic activity, is observed, judging by the far UV circular dichroism of the protein. This process of partial refolding can easily be followed using DAPI as an extrinsic reporter group, able to monitor the kinetics of formation and decay of a highly fluorescent intermediate. This process becomes faster at a lower pH, at least in the limited range investigated (pH 4.5-5.5), in which the refolded protein does not aggregate, but, in contrast to unfolding, is almost independent in ionic strength. PMID- 9397521 TI - Salt and pH effects on electrochemistry of myoglobin in thick films of a bilayer forming surfactant. AB - Salt concentration and pH of external solutions were shown to control the electrochemistry of the heme protein myoglobin (MbFe(III)-H2O) in stable, ordered films of didodecyldimethylammonium bromide (DDAB). Protonation of aquometmyoglobin (MbFe(III)-H2O) in these films precedes electron transfer from electrodes, causing formal potentials to shift negative as pH increases from 5 to 8. At pH > 8, MbFe(III)-H2O dissociates to MbFe(III)-OH, which is reduced directly at the electrode at higher rates than MbFe(III)-H2O. Correlations of voltammetric data with FT-IR spectra suggested that at pH < 4.6, an unfolded form of Mb resides in the films and is reduced directly. The concentration of salt in solution influences electrochemical properties of Mb-DDAB films by its influence on Mb conformation and by effects on interfacial Donnan potentials. NMR indicated strong binding of anions to Mb within DDAB films. Bound anions may neutralize positive charge on Mb's surface so that it can reside in a partly hydrophobic environment, as postulated on the basis of previous ESR and linear dichroism studies. PMID- 9397522 TI - Effect of inhalation anaesthetics on the phase behaviour, permeability and order of phosphatidylcholine bilayers. AB - We have used differential scanning calorimetry and fluorescence anisotropy measurements to investigate the effect of five inhalation anaesthetics of diverse chemical structure (halothane, enflurane, n-pentane, chloroform and diethylether) on the phase behaviour of liposomes prepared from dimyristoylphosphatidylcholine (DMPC) and dipalmitoylphosphatidylcholine (DPPC), respectively. The incorporation of these anaesthetics induced a decrease of the phase transition temperature and/or a broadening of the phase transition peak depending on the transverse localisation of the investigated anaesthetic. At high anaesthetic concentrations we observed the disappearance of the pretransition peak and the appearance of a shoulder on the main phase transition peak due to the domain formation of the anaesthetics. An anaesthetic induced carboxyfluorescein efflux from the vesicle lumen was completed within a few minutes after the addition of the anaesthetics, probably resulting from a transient formation of membrane holes. All results are discussed with regard to the physicochemical properties of the anaesthetics applied. PMID- 9397523 TI - Melanotropic peptides-lipid bilayer interaction. Comparison of the hormone alpha MSH to a biologically more potent analog. AB - The interaction of the native peptide alpha-melanocyte stimulating hormone (alpha MSH) and the biologically more active analog [Nle4, D-Phe7]-alpha-MSH(MSH-I) with lipid vesicles was studied by spin label electron spin resonance (ESR) spectroscopy and circular dichroism (CD). Using spin labels located at the membrane interface and at different depths along the acyl chain, it was shown that the binding of both peptides to the membrane induces tighter lipid packing at all the monitored positions. However, the effect of the analog on the spin label ESR parameters was much more evident, and suggested that it penetrates farthest into the lipid matrix than the native molecule. Lipid partition coefficients were calculated based on the effect the peptides cause on the ESR spectra of spin labels incorporated in the membrane. For the biologically more potent peptide, the partition coefficient was found to be about 4-times greater than that of the native hormone. For the same concentration of peptide bound to the membrane, MSH-I was found to cause a slightly greater effect on the membrane structure than alpha-MSH, in accord with its possible deeper penetration into the bilayer. CD spectra in aqueous solution and in the alpha-helix inducing solvent 2,2,2-trifluoroethanol showed that the two peptides have somewhat different structures in solution, though similar conformational changes occur in both peptides as a result of their interaction with negatively charged vesicles or micelles. The higher peptide-lipid association constant and the deeper penetration of the analog into lipid bilayers could be related to its greater activity and/or prolonged action. PMID- 9397524 TI - Multiple nucleosome positioning with unique rotational phasing on multimers of the light-responsive elements of pea rbcS-3A and rbcS-3.6 genes: comparison between experimental and theoretical mapping. AB - Nucleosome positioning along two DNA tracts, corresponding to tetramers of the light-responsive elements of pea rbcS-3A and rbcS-3.6 genes, were studied by experimental (exonuclease III mapping and band shift electrophoresis) as well as theoretical methods. Multiple nucleosome positioning with unique rotational phase was derived from both methods in satisfactorily good agreement, if nucleosome dyad axis positions are considered. Theoretical and experimental distributions of nucleosome frequencies appear different, probably on account of DNA sequence dependent digestion kinetics of exonuclease III. PMID- 9397525 TI - Delayed dissociation of in vitro moving actin filaments from heavy meromyosin induced by low concentrations of Triton X-100. AB - The in vitro motility of fluorescent actin filaments over heavy meromyosin (HMM) was studied in the presence of the nonionic detergent Triton X-100. Below 0.004% Triton X-100 concentration, motility was not affected. Above 0.007%, motility was not observed because actin filaments were dissociated from HMM. In the Triton X 100 concentration range of 0.004-0.007%, the sliding actin filaments dissociated from HMM with a delay. The dissociation delay time decreased with increasing Triton X-100 concentration, increasing ATP (adenosine-5'-triphosphate) concentration, and increasing temperature. The delayed acto-HMM dissociation was absent when weak-binding kinetic intermediates of the myosin ATPase cycle (M.ATP and M.ADP-Pi) were used. The presence of sliding movement was necessary to evoke the delayed acto-HMM dissociation. The acto-HMM dissociation delay was independent of actin filament length. For a given Triton X-100 concentration, the dissociation delay time was found to be inversely proportional to sliding velocity, indicating that actin filaments travel a more or less constant distance prior to dissociation from HMM. The actin-activated HMM ATPase activity was not inhibited by Triton X-100; rather, it was slightly enhanced. The results imply the presence of a motility-associated conformational change in acto-HMM. PMID- 9397526 TI - Molecular acoustic as a new tool for the study of biophysical properties of lipoproteins. AB - The method of measurement of velocity and absorption of ultrasound at a fixed frequency (7.2 MHz) and measurement of density were used to study the physical properties of high- (HDL3) and low- (LDL) density lipoproteins. We found substantial changes in velocity number [u] and absorption number [alpha lambda] on temperature, which reflect structural changes in the hydrophobic core of LDL at the thermotropic-phase transition. The absorption number revealed broad changes in temperature for both classes of lipoproteins (LP). The density of LP also depends on temperature but in considerably less degree than the acoustic parameters. The values of acoustic parameters were determined, showing that LDL and HDL3 greatly differ with respect to adiabatic compressibility. PMID- 9397527 TI - The reaction of oxygen with radicals from oxidation of tryptophan and indole-3 acetic acid. AB - The oxidation of tryptophan and indole-3-acetic acid (IAA) by the dibromine radical anion or peroxidase from horseradish in aqueous solution was investigated and compared, especially with respect to the involvement of oxygen and superoxide. Using EPR with spin-trapping, the tryptophanyl radical, generated by either method was found to react with oxygen, although this reaction is too slow to be observed by pulse radiolysis (k < 5 x 10(6) dm3 mol-1 s-1). No superoxide results from this reaction, thus excluding an electron-transfer mechanism and suggesting the formation of a tryptophan peroxyl radical, possibly in a reversible process. These observations imply that in proteins where the tryptophanyl radical exists as a stable species it must either have its reactivity modified by the protein environment or be inaccessible to oxygen. The related molecule LAA is oxidized by either peroxidase or Br2.- to a radical cation that decarboxylates to yield a skatolyl radical. The latter reacts with oxygen to give a peroxyl radical that does not release superoxide. However, O2.- is formed during the peroxidase-catalyzed oxidation of indoleacetic acid. This supports the hypothesis that the peroxidase can act in an oxidase cycle involving ferrous enzyme and compound III, with superoxide as a product. PMID- 9397528 TI - Surface electric properties of thylakoid membranes from Arabidopsis thaliana mutants. AB - Electric light scattering measurements of thylakoid membranes from wild type and two mutant forms (JB67 and LK3) of Arabidopsis thaliana have shown that application of external electric pulses induces electric dipole moments of different origin. The asymmetric surface charge distribution and electric polarizability are significantly altered by the lipid modification. Mild trypsin treatment of Arabidopsis thylakoids leading to digestion of small polypeptides from the light-harvesting chlorophyll a/b protein complex of photosystem II (LHCP II) gives evidence for a lower content of LHCP II in the mutant forms. The results demonstrate the significance of the level of thylakoid lipid unsaturation in determining the surface charge distribution through changes either in the pigment-protein content and membrane appression induced by the lipid modification or in the exposure of charged polypeptides on the thylakoid membrane surface(s) arising from alteration of the lipid geometry. PMID- 9397529 TI - Role of phosphatidylethanolamine lipids in the stabilization of protein-lipid contacts. AB - We have investigated the effect of lipids with phosphatidylethanolamine (PE) head groups on the stabilization of contacts between the tryptophan side chains of gramicidin and the lipid head groups. We initially developed two fluorescence methods that can be correlated to the spontaneous curvature of DOPC/DOPE and DOPC/DOPEme. One is based on bilayer structure and measures the rotational motion of a probe located close to the membrane surface relative to a more deeply-buried probe. The second is based on surface hydration/polarity and measures the emission energy of a polarity-sensitive probe located on the membrane surface. We used these methods to estimate the pseudo-curvature (i.e., curvature obtained by fluorescence measurements) of lipids with dimyristyl chains, and their pressure and temperature dependence. We then investigated the stability of gramicidin tryptophan-lipid contacts in DMPC/DMPE as a function of temperature and pressure. Stability was assessed by tryptophan rotational motion as determined by fluorescence anisotropy, since rotational motion is limited when the indoles are hydrogen bonded to the lipid head groups. The results suggest that the presence of PE lipids destabilizes these contacts due to either their smaller size relative to PC head groups, or their tendency to self-interact. Fluorescence quenching studies support these results. PMID- 9397530 TI - Dynamic light scattering study of fine semiflexible fibrin networks. AB - Fine fibrin networks have been investigated using the dynamic light scattering (DLS) technique. At the shortest delay times, t, the dynamic structure factor s(q,t) is found to depend on time according to an exponential function and, at intermediate delay times (up to 1 ms), to a stretched exponential. At longer times (t > 1 ms), a progressively increasing deviation from the stretched exponential behaviour has been observed. These results are in agreement with the theoretical predictions of a recently forwarded model for semiflexible polymers in semidilute solutions [K. Kroy and E. Frey, Physical Review E 55 (1996) p. 3092.], despite the fact that fibrin networks are made up of crosslinked branched polymers. The model, moreover, allows the calculation from the initial decay rate gamma q(0) of the average diameter of the fibrin fibres, a. The value of a = 30 +/- 2 nm, at fibrinogen concentration c(f) = 1676 nM and ionic strength 0.5, fits well into the data reported in electron microscopy studies. A concentration dependence of the average diameter of the fibrin fibres has been observed which saturates at the highest concentrations. The diameter of fibrin fibres is an important component in determining the physical properties of the fibrin networks, since the radial growth of fibrin fibres is limited by twisting during protofibrils aggregation. Our results indicate the importance of taking into account intrinsic semiflexibility in studying the physical properties of 'real' polymers and emphasize the high sensitivity of the DLS technique to investigate biological polymers also at the lowest concentrations where the systems are very fragile. PMID- 9397531 TI - The molecular biology of secreted enzyme production by fungi. AB - Enzymes from filamentous fungi are already widely exploited, but new applications for known enzymes and new enzymic activities continue to be found. In addition, enzymes from less amenable non-fungal sources require heterologous production and fungi are being used as the production hosts. In each case there is a need to improve production and to ensure quality of product. While conventional, mutagenesis-based, strain improvement methods will continue to be applied to enzyme production from filamentous fungi the application of recombinant DNA techniques is beginning to reveal important information on the molecular basis of fungal enzyme production and this knowledge is now being applied both in the laboratory and commercially. We review the current state of knowledge on the molecular basis of enzyme production by filamentous fungi. We focus on transcriptional and post-transcriptional regulation of protein production, the transit of proteins through the secretory pathway and the structure of the proteins produced including glycosylation. PMID- 9397532 TI - Human gene therapy. AB - Human gene therapy and its application for the treatment of human genetic disorders, such as cystic fibrosis, cancer, and other diseases, are discussed. Gene therapy is a technique in which a functioning gene is inserted into a human cell to correct a genetic error or to introduce a new function to the cell. Many methods, including retroviral vectors and non-viral vectors, have been developed for both ex vivo and in vivo gene transfer into cells. Vectors need to be developed that efficiently transfer genes to target cells, and promoter systems are required that regulate gene expression according to physiologic needs of the host cell. There are several safety and ethical issues related to manipulating the human genome that need to be resolved. Current gene therapy efforts focus on gene insertion into somatic cells only. Gene therapy has potential for the effective treatment of genetic disorders, and gene transfer techniques are being used for basic research, for example, in cancer, to examine the underlying mechanism of disease. There are still many technical obstacles to be overcome before human gene therapy can become a routine procedure. The current human genome project provides the sequences of a vast number of human genes, leading to the identification, characterization, and understanding of genes that are responsible for many human diseases. PMID- 9397533 TI - Alginate production by Azotobacter vinelandii. AB - Although all commercial alginates are today of algal origin, there is interest in the production of alginate-like polymers from bacteria. The species Azotobacter vinelandii seems to be the best candidate for the industrial production of alginate molecules characterized by a chemical composition, molecular mass and molecular mass distribution suited to a well defined application, especially required in the biotechnological, biomedical and pharmaceutical fields. The production of alginate by A. vinelandii has been to date widely investigated both in batch (mainly in the shaken flask scale) and in continuous cultures. This article summarizes current knowledge on the structure and properties of alginates and their applications and presents an overview of up-dated research on the physiology, genetics and kinetics of the production of alginate by Azotobacter vinelandii and its rheology, including the results of our recent studies. PMID- 9397534 TI - Consequences of disrupting the gene that encodes alpha-glucosidase II in the N linked oligosaccharide biosynthesis pathway of Dictyostelium discoideum. AB - We have identified and disrupted the gene coding for alpha-glucosidase II in Dictyostelium discoideum. This enzyme is responsible for removing two alpha 1,3 linked glucose residues from N-linked oligosaccharides on newly synthesized glycoproteins. Mutagenesis by restriction enzyme-mediated integration (REMI) generated a clone, DG1033, which grows well but forms abnormal fruiting bodies with short, thick stalks. The strain lacks alpha-glucosidase II activity and makes incompletely processed N-linked oligosaccharides that are abnormally large and have fewer sulfate and phosphate esters. The morphological, enzymatic, and oligosaccharide profile phenotypes of the disruption mutant are all recapitulated by a targeted disruption of the normal gene. Furthermore, all of these defects are corrected in cells transformed with a normal, full-length copy of the gene. The phenotypic characteristics of DG1033 as well as chromosomal mapping of the disrupted gene indicate that it is the site of the previously characterized modA mutation. The Dictyostelium gene is highly homologous to alpha-glucosidase II genes in the human and the pig, C. elegans, and yeast. Although various cell lines have been reported to be defective in alpha-glucosidase II activity, disruption of the Dictyostelium gene gives the first example of a clear developmental phenotype associated with loss of this enzyme. PMID- 9397535 TI - Differential expression of gap junctions in neurons and astrocytes derived from P19 embryonal carcinoma cells. AB - The P19 embryonal carcinoma cell line represents a pluripotential stem cell that can differentiate along the neural or muscle cell lineage when exposed to different environments. Exposure to retinoic acid induces P19 cells to differentiate into neurons and astrocytes that express similar developmental markers as their embryonic counterparts. We examined the expression of gap junction genes during differentiation of these stem cells into neurons and astrocytes. Untreated P19 cells express at least two gap junction proteins, connexins 26 and 43. Connexin32 could not be detected in these cells. Treatment for 96 hr with 0.3 mM retinoic acid induced the P19 cells to differentiate first into neurons followed by astrocytes. Retinoic acid produced a decrease in connexin43 mRNA, protein, and functional gap junctions. Connexin26 message was not affected by retinoic acid treatment. The neurons that developed consisted of small round cell bodies extending two to three neurites and expressed MAP2. Connexin26 was detected at sites of cell-cell and cell-neurite contact within 3 days following differentiation with retinoic acid. The astrocytes were examined for production of their intermediate filament marker, glial fibrillary acidic protein (GFAP). GFAP was first detected at 8 days by Western blotting. In culture, astrocytes co-expressed GFAP and connexin43 similar to primary cultures of mouse brain astrocytes. These results suggest that differentiation of neurons and glial cells involves specific connexin expression in each cell type. The P19 cell line will provide a valuable model with which to examine the role gap junctions play during differentiation events of developing neurons and astrocytes. PMID- 9397536 TI - Characterization of a family of repetitive sequences that is eliminated late during macronuclear development of the hypotrichous ciliate Stylonychia lemnae. AB - A repetitive element from the hypotrichous ciliate Stylonychia lemnae was characterized by restriction and hybridization analysis. This repetitive element is present in about 5,000-7,000 copies per haploid genome in the micronucleus and the macronuclear anlagen. Its DNA sequence is very conserved, but the length of the repetitive sequence blocs is variable. In some cases, it is associated with telomeric sequences and macronucleus-homologous sequences. Restriction analysis of genomic micronuclear and macronuclear anlagen DNA and in situ hybridization showed that the repetitive sequences are amplified during the formation of polytene chromosomes. They are localized in many bands of the polytene chromosomes and are eliminated during the degradation of the polytene chromosomes. Possible functions of the repetitive sequences during macronuclear differentiation are discussed. PMID- 9397537 TI - Identification of a growth arrest specific (gas 5) gene by differential display as a candidate gene for determining susceptibility to hyperthermia-induced exencephaly in mice. AB - Neural tube defects (NTDs) are among the most common congenital malformations, affecting approximately 1 per 1,000 liveborn infants in the United States [Nakano, 1973; Richards et al., 1972]. Maternal exposure to hyperthermia, either through recreational sources or due to an infectious agent, is thought to account for approximately 10% of observed NTD cases. The specific genes conferring susceptibility or resistance to hyperthermia-induced NTDs have not been identified. This study used differential display-polymerase chain reaction (DD PCR) to characterize alterations in gene expression in the anterior embryonic neural tube of two highly inbred murine strains (SWV/Fnn, LM/Bc/Fnn) known to differ in their genetically determined susceptibility to heat-induced NTDs. Herein, we report the neural tube-specific differential expression of the growth arrest specific (gas 5) gene in the highly susceptible SWV/Fnn strain during neural tube closure (NTC). Although the expression of gas 5 did not appear to be altered by the teratogenic heat treatment, its spatial and strain-specific pattern of expression makes it an excellent candidate gene responsible for the observed genetic differences in NTD susceptibility between these two inbred murine strains. PMID- 9397538 TI - Molecular characterization of a heat shock cognate cDNA of zebrafish, hsc70, and developmental expression of the corresponding transcripts. AB - To elucidate the potential role of the hsp70 gene family in developmental processes in vertebrates, we chose to study the expression of one of these genes in zebrafish. A zebrafish gastrula cDNA library was screened with a Pleurodeles waltl hsp70 cDNA probe. A 2.3-kb cDNA was thus isolated and sequenced. The predicted amino acid sequence contained an open reading frame encoding for a 649 amino acid polypeptide. Sequence analysis showed strong homology with hsp70 related gene sequences in other species; in particular, the strongest homology was found with the cognate members of this family. Tests of heat inducibility revealed that transcripts were expressed at normal temperature, but the level of transcript expression increased after heat shock. Moreover, experiments of the neosynthesis of total proteins in heat shock conditions and corresponding immunoblotting assays showed that 24-h-stage embryos are able to respond to heat shock. The quantity of 70 kDa proteins, recognized by a specific antibody of the HSP/C70 protein family, is expressed in control condition and increased significantly after heat shock. Furthermore, Northern blot analysis of transcript expression showed that the corresponding mRNAs were detected throughout embryonic development in the absence of any heat shock. Our clone, named hsc70, thus corresponded to a cognate member of the hsp70 gene family, expressed under normal conditions during development, but also heat inducible. The spatio-temporal pattern of transcripts during development was determined by in situ hybridization on wholemount embryos at different stages. As a maternal RNA, hsc70 mRNA was uniformly present in the embryo, up to the end of gastrulation. Later, a tissue specific enrichment of hsc70 transcripts was detected in the central nervous system (CNS) and in a fraction of the somites. These results suggest that the hsc70 gene may be involved in developmental differentiation events. PMID- 9397539 TI - Three subsets of genes whose tissue specific expression is sex and age-dependent can be identified within the rat alpha 2u-globulin family. AB - The rat alpha 2u-globulins are encoded by a multigene family whose 20-25 members are subjected to multihormonal regulation that is dependent upon the sex of the animal, the developmental stage and the tissue being examined. Using RT-PCR and diagnostic restriction analysis of the products, we have examined the specificity of the expression of different members of the gene family. All family members can be classified into three subsets, depending on how the amplified cDNA responds to digestion with ApaLI, SstI and VspI. Subset A contains the restriction sites for both ApaLI and SstI but not VspI and typifies the genes expressed in the salivary glands of both mature and juvenile animals of both sexes, where it is the only subset expressed. This subset of genes also accounts for all the transcripts observed in the kidneys and mammary glands of juvenile males. Although subset A was represented in the transcript populations of all the other tissues examined, its proportion relative to the total varied greatly. The two other subsets were subset V, which contains only the restriction site for VspI, and subset N, which lacks all three restriction sites. In all the other tissues examined, two or all three of the subsets were expressed, usually in a manner that was unique to the sex and age of the tissue in question. The proportion of each of the three alpha 2u-globulin subsets in the alpha 2u-globulin gene family was determined by quantitation of the restriction products of amplified genomic DNA. Interestingly, the most prevalent subset in the genome (N) has the most limited tissue expression pattern, but is found in liver and preputial glands, the tissues expressing the most substantial quantities of alpha 2u-globulin. These results indicate the complexity of the regulation of the alpha 2u-globulins and point to the necessity for gene specific analyses if the expression of the family is to be understood in molecular terms. PMID- 9397540 TI - Hyphenated liquid chromatographic method for the determination of colistin residues in bovine tissues. AB - A selective and sensitive high-performance liquid chromatographic (HPLC) method has been developed for the measurement of colistin residues in milk and in four bovine tissues (i.e., muscle, liver, kidney, and fat). The sample treatment consists of protein precipitation using 10% (w/v) trichloroacetic acid, solid phase purification on C18 cartridges, and precolumn derivatization of colistin with ortho-phthalaldehyde and 2-mercaptoethanol in borate buffer (pH 10.5). This latter step is performed automatically, and the resulting reaction mixture is injected into a switching HPLC system including a precolumn and an analytical column packed with end-capped LiChrospher RP18 (5 microns). Washing the precolumn and final elution onto the analytical column are conducted using acetonitrile 0.01M phosphate buffer (pH 7.0) mixtures with respective proportions of 65:35 and 68:32 (v/v). Detection is carried out by spectrofluorometry (excitation wavelength, 340 nm; emission wavelength, 440 nm). The retention times of the derivatives corresponding to the two main components of colistin (i.e., polymyxins E2 and E1) are approximately 14 and 18 min, respectively. The structural study of the derivatives corresponding to polymyxins E1 and E2 is carried out by HPLC coupled with electrospray mass spectrometry; data obtained confirms that the derivatization process occurs with the five amino groups of the analytes. Selectivity is obtained in the HPLC system versus other coadministered anti-infective drugs (beta-lactams, aminoglycosides, tetracyclines, and sulphonamides) and endogenous compounds. Quantitation is performed using the sum of the peak areas of polymyxin E1 and polymyxin E2 derivatives. Testing linearity affords correlation coefficients greater than 0.990 for calibration curves in the range of 10-500 microL/L for milk, 50-1000 micrograms/kg for muscle and fat, and 100-1000 micrograms/kg for kidney and liver. Relative standard deviation values are less than 10% at a concentration of 25 micrograms/L in milk and 100 micrograms/kg in tissues (six replicates); recoveries are higher than 60%. PMID- 9397541 TI - Determination of phloxine B and uranine in water by capillary zone electrophoresis. AB - Phloxine B and uranine are color additives in drugs and cosmetics as well as potential photoactive insecticides. A capillary zone electrophoretic (CZE) method is developed to determine phloxine B and uranine in water. A fused-silica capillary (67 cm, 75-micron i.d.) and borate buffer are used. Migration of phloxine B and uranine increases slightly as the pH of the running buffer increases between the range of 8-9. Although there are only slight effects of ionic strength on the analyte migration in the range of 0-20 mM NaCl in the running buffer, the migration of phloxine B and uranine increases as the percentage of methanol in the samples increases. Methanol shows little effect on the quantitation of phloxine B and uranine. The CZE procedure is applied to determine phloxine B and uranine fortified in tap and stream water samples. Solid phase extraction is employed to recover the analytes spiked in the water samples. Recoveries range from 87-112% for phloxine B spiked at 10-200 ppb in the tap and stream water. Uranine recoveries are 86-91% at fortification levels of 10-50 ppb in the water samples. PMID- 9397542 TI - High-performance liquid chromatographic determination of 2-furaldehyde and 5 hydroxymethyl-2-furaldehyde in fruit juices. AB - A method for the determination of 2-furaldehyde (F) and 5-hydroxymethyl-2 furaldehyde (HMF) in fruit juices by high-performance liquid chromatography (HPLC) is described. The method is based on the formation of the 2,4 dinitrophenylhydrazones of carbonyl compounds and subsequent separation of these derivatives. Derivatization is carried out by utilizing an acidic solution of 2,4 dinitrophenylhydrazine in acetonitrile. Precipitation of the derivatives of carbonyl compounds is thus avoided, and direct injection of the sample into the HPLC system is allowed. The procedure offers a high specificity and a detection limit of the order of 10(-8) mol/L. Recoveries of 95-98% are obtained from apple juice spiked at different levels with both analytes. The reproducibility (mean of six determinations) is +/- 2% for F and +/- 3% for HMF. PMID- 9397543 TI - Injuries due to letter bombs. AB - In Austria in late 1993 ten letter bombs were sent to outstanding persons who have been engaged in the care of foreigners. Four of these bombs detonated, when they were opened by the addressee. The remaining six bombs were discovered in time and could be deactivated by specialists. The construction of these bombs and the lesions sustained by the four victims will be discussed. The injuries mainly concerned the left hand, i.e., the hand used by right-handed persons to hold a letter when opening it. The way holding the letter was of crucial influence on the degree of injury, as with the same explosive charge (which can be assumed deducing from the investigation of the deactivated bombs) injuries varied considerably. They ranged from minor tissue-lesions to mutilated fingers and the risk of exsanguination. PMID- 9397544 TI - Extraction of single-copy nuclear DNA from forensic specimens with a variety of postmortem histories. AB - Specimens of human bone, teeth and dried blood spots from 3 months to 91 years old, with a variety of postmortem histories, were used in a comparative study of recovery of single-copy nuclear DNA sequences from forensic material. Sequences of the amelogenin and HLA-DPB1 genes were chosen for their value in sexing and identification. Sequences of the mitochondrial non-coding region V were also amplified to compare the recovery of mitochondrial and single-copy nuclear DNA. A variation of the silica method for DNA extraction was refined for application to the forensic specimens in this sample. Single-copy nuclear DNA was amplified from 100% of recent postoperative bone specimens (n = 6), 80% of forensic teeth and bone specimens (n = 10), 78% of recently extracted teeth (n = 18), 78% of exhumed bone up to 91 years old (n = 37) and 69% of 15 year old bone specimens fixed in 10% formalin (n = 20). Amelogenin sexing was correct in 85% of cases (n = 74) in which the sex of the donor had been recorded. There was no correlation between the age of the specimen and the extent of DNA preservation. PMID- 9397545 TI - Genetic individualization of domestic cats using feline STR loci for forensic applications. AB - A group of ten short tandem repeat (STR) loci suitable for PCR typing from DNA of domestic cats is evaluated for genetic individualization using blinded samples of eight putative feline blood specimens. The ten loci were also typed in a 70 member cat pedigree to demonstrate Mendelian inheritance and independent assortment. A "match window" or measurement precision estimate was empirically established by determining the maximum gel migration difference among alleles identical by descent in different individuals of the pedigree. Hardy-Weinberg equilibrium and abundant heterozygosity was observed for each locus in cat population samples from Canada and the USA. The probabilities of two unrelated individuals matching by chance (Pm) at all ten loci was estimated as 1.35 x 10( 10). We present a conservative approach to compute, for forensic consideration, the mathematical likelihood of a chance genotypic match between DNA evidence from a crime scene and the suspect composite STR genotypes for species or populations when genotype frequency information is not available. PMID- 9397546 TI - Violent behaviors associated with the antichrist delusion. AB - Delusions involving the antichrist concept have been occasionally reported. Some cases of the antichrist delusion have been associated with violent behavior. In this article we describe the case of a man who suffered from a chronic antichrist delusion and who also displayed repeated and serious violent behaviors. We also discuss the role of the antichrist delusion as well as other psychotic symptoms in the genesis of violence in the present case. PMID- 9397547 TI - Sex estimation from the metatarsals. AB - Discriminant-function analysis of osteometric data from the metatarsals of 200 individuals in the Terry Collection provides a reliable method for estimating sex. Functions derived from individual metatarsals and from complete sets of metatarsals are tested on the sample used to generate the functions and on two independent samples: one comprising 25 additional individuals from the Terry Collection and the other comprising 12 cadavers donated to the University of Missouri, Functions based on race-specific samples (blacks and whites) and on the pooled-race sample correctly classify 83 to 100% of each sample (including a jackknifed study sample), with a few exceptions. These results are similar to sex estimation methods from other regions of the appendicular skeleton. PMID- 9397548 TI - HIV seroprevalence rates among homicide victims in New York City: 1991-1993. AB - This study assessed HIV seroprevalence in homicide victims killed in New York City in 1991-1993, using data from the Office of Chief Medical Examiner. Among 5852 homicide victims there were 344 (5.9%) victims who were HIV positive. Females were just as likely as males to be HIV positive. For females, the highest rates were in the 25-34 year (11.7%) and 35-44 year (12.6%) age categories. For males the highest rates were in the 35-44 year (13.7%) and 45-54 year (11.5%) age categories. Other than there being no HIV positive Asian victims, there were no differences in HIV rates among racial/ethnic groups. The highest rates of HIV infection for homicide victims were among those using both opiates and cocaine (males: 23.0%; females: 27.3%). Women, not men, using cocaine alone had a high HIV positive rate (18.4%). Victims not using these drugs had rates of HIV around 2%. The authors believe that the high risk of HIV among homicide victims, may be due to the use of cocaine and associated risky use of needles and risky sex practices. PMID- 9397549 TI - Midge larvae (Diptera: Chironomidae) as indicators of postmortem submersion interval of carcasses in a woodland stream: a preliminary report. AB - Data on colonization of rat carcasses by aquatic insects in riffle and pool areas of a small woodland stream were obtained to elucidate patterns potentially useful for determining the postmortem submersion interval of corpses in flowing water habitats. After 39 days, the carcasses had no visual signs of deterioration in the absence of large scavenging animals. Midge larvae (Diptera: Chironomidae) were the dominant insects colonizing the carcasses. No patterns in numbers of larvae over time were evident, but the diversity of genera increased after 29 days in the riffle. Also, Orthocladius larvae did not begin to colonize the carcasses until after 13 days of submersion in the riffle and after 20 days of submersion in the pool. Although separated only by 20 m, the riffle and pool rats had dissimilar faunal assemblages. This suggests that different indices for determining the postmortem submersion interval of corpses based on midge larvae colonization should be developed for these two habitats. This investigation does not provide replicated data, but does shed light on what may happen to mammalian carcasses placed in a stream at a particular time of the year. PMID- 9397550 TI - The response of the Intoxilyzer 5000 to five potential interfering substances. AB - A study was conducted of potential vapor phase interferents which could be present on human breath and also be capable of inducing a false-positive response for ethanol on the evidential infrared-based breath testing device, the Intoxilyzer-5000. This involved preparation and validation of a range of vapor standards, which were introduced to the instrument using a dynamic flow double bubbler system. Potential interferents were chosen on the basis of both their infrared signatures and their general availability, and included toluene, m xylene, o-xylene, methanol and isopropanol. All compounds tested were found to be capable of inducing false-positive readings for ethanol in a highly reproducible manner, as a result of which it has been possible to derive precise least-squares equations describing the ethanol readout expected for any given blood concentration of toluene, m-xylene, o-xylene, methanol and isopropanol. The likelihood of an interference compromising the integrity of the analysis is related to both the toxicological significance and prevalence of a given blood concentration of each solvent, and the point at which the instrumental interference light is triggered in each case. PMID- 9397551 TI - Concentration-time profiles of ethanol in arterial and venous blood and end expired breath during and after intravenous infusion. AB - Ethanol (0.40 g/kg) was administered to 13 healthy men by intravenous (i.v.) infusion at a constant rate for 30 min. The concentrations of ethanol in arterial blood (ABAC), venous blood (VBAC), and end-expired breath (BrAC) were measured at 17 exactly timed intervals. Blood-ethanol was determined by headspace gas chromatography and breath-ethanol was measured with a quantitative infrared analyzer (DataMaster). BrAC was multiplied by 2300 to estimate the concentrations of alcohol in blood. During the infusion of ethanol, ABAC exceeded VBAC by about 10 mg/dL on the average and ABAC was also higher than BrAC x 2300 by about 4 mg/dL on average. When infusion of alcohol ended, ABAC, VBAC, and BrAC were 94.8 +/- 2.06 (+/- SE), 84.7 +/- 1.54, and 89.3 +/- 2.10 mg/dL, respectively. The concentrations of alcohol in blood (ABAC and VBAC) and breath decreased abruptly after the administration of alcohol stopped and by 5 min postinfusion, the A-V differences in concentration of ethanol were small or negligible. The mean apparent half-life of the distribution plunge was 7 to 8 min, being about the same for ABAC, VBAC, and BrAC. The disappearance rate of ethanol was 15.5 +/- 0.55 mg/ dL/h (mean +/- SE) for arterial blood, 15.2 +/- 0.49 mg/dL/h for venous blood, and 16.3 +/- 0.73 mg/230 L/h for breath; no significant differences were noted (p > 0.05). We conclude that A-V differences in the concentration of ethanol exist during the loading phase but are rapidly abolished when the administration of ethanol terminates. In the post-absorptive phase of ethanol kinetics, when alcohol has mixed with the total body water, VBAC exceeds ABAC by about 1-2 mg/100 mL on average. PMID- 9397552 TI - Influence of pigmentation on the codeine content of hair fibers in guinea pigs. AB - Tortoise shell guinea pigs (n = 7) were administered codeine (1 mg/mL codeine base) in their drinking water for 3 weeks. Black, reddish-brown and white hair was collected separately from each animal before and after treatment. The hair samples were analyzed by GC/MS. The experiment showed positive results for all hair fibers with large individual variability of drug incorporation. Low drug intake resulted in small differences of the drug content in hair fibers different in color, whereas in cases of high drug intake a strong influence of hair pigmentation on the analytical results was observed. The highest drug content was always found in black hair samples, non-pigmented hair showed the lowest drug concentrations and the drug content in reddish-brown fibers was less than in black hair samples from the same animal. From the results it was concluded, that eumelanins rather than phenomelanins are the decisive factor for codeine-melanin binding in hair and the amount of drug intake was suggested to determine the relevance of hair pigmentation on the analytical results. PMID- 9397553 TI - Ethyl glucuronide concentration in serum of human volunteers, teetotalers, and suspected drinking drivers. AB - The kinetic profile of ethanol and ethyl glucuronide (EtG) in serum was investigated in three subject groups: 1) Healthy, moderately drinking volunteers (daily intake less than 30 g ethanol) who ingested a single dose of ethanol. In this group the maximum of serum ethyl glucuronide concentration (SEtGC) and of serum ethanol concentration (SEC) did not exceed 3.7 mg/L and 1.5 g/L respectively. EtG peaked 2 to 3.5 h later than ethanol. EtG was eliminated with a terminal half-life of 2 to 3 h. EtG decreased slower than ethanol--the metabolite could still be determined in serum up to 8 h after complete ethanol elimination. 2) In serum samples of teetotalers neither ethanol nor EtG could be found. 3) In 37 of 50 serum samples of drivers suspected of driving under the influence of ethanol, SEtGC was found between the limit of detection (0.1 mg/L) and 20 mg/L. If the SEC is less than 1 g/L and the SEtGC is significantly higher than 5 mg/L, we assume alcohol misuse. PMID- 9397554 TI - Who does the family court refer for psychiatric services? AB - Demographic differences between adolescents referred for psychiatric services by the Family Court and by facility staff at a state-run juvenile justice evaluation center are examined. Those groups are then compared to the facility's general population. It is concluded that race, gender, age, and judicial discretion are the factors that distinguish court-referred adolescents from their counterparts referred by facility staff and in the general population. PMID- 9397555 TI - Forensic analysis and psycholegal implications of parricide and attempted parricide. AB - In a retrospective archive study, 64 adjudicated adult cases involving the murder or attempted murder of at least one parent, referred for forensic evaluations are described. Biographic, demographic, diagnostic, crime scene, psycholoegal opinion, and disposition data are presented. Results indicated a 40% rate of insanity acquitees. Attempted parricide subjects were more likely to have inpatient psychiatric histories, witnesses present during the criminal act, nonresponsiveness towards their actions, competency raised, and a hospital disposition. Gender and ethnicity were found to have a significant effect on ultimate disposition. Fifty-four percent of the sample opined psychotic were sentenced to prison, suggesting other factors considered by judge and jury. Profile characteristics and typologies are presented. The findings are compared to studies involving parricide and legal strategies involving similar cases. PMID- 9397556 TI - A pilot study using the first cervical vertebra as an indicator of race. AB - The articular surfaces and vertebral foramen of the first cervical vertebra can be used to estimate race from complete and fragmentary specimens. Eight measurements taken from 200 vertebrae from the Terry and Hamann-Todd collections (Smithsonian Institution and Cleveland Museum of Natural History, respectively) were used to construct 13 discriminant functions that predict race with 76-60% accuracy. PMID- 9397557 TI - Comparison of three DNA extraction methods on bone and blood stains up to 43 years old and amplification of three different gene sequences. AB - Extraction of amplifiable DNA-from degraded human material in the forensic context remains a problem, and maximization of yield and elimination of inhibitors of the Polymerase Chain Reaction (PCR) are important issues which rarely feature in comparative studies. The present work used PCR amplification of three DNA sequences (HLA DPB1, amelogenin and mitochondrial) to assess the efficiency of three methods for extracting DNA (sodium acetate, magnetic beads and glass-milk) from 32 skeletal samples and 25 blood stains up to 43 years old. The results, analyzed using multivariate statistics, confirmed that the extraction method was crucial to the subsequent detection of amplification products; the glass-milk protocol performed better than sodium acetate, which was better than magnetic beads. Successful amplification also depended on gene sequence, multiple copy mitochondrial sequences performing best; however, with the singly copy sequences, the longer HLA DPB1 (327 bp) being detected just as often as the shorter amelogenin (106/112 bp). Amplification products were obtained more frequently from blood stains than bone, perhaps reflecting differences inherent in the material, and from younger compared with older specimens, though plateauing seemed to occur after 10 years. PCR inhibitors were more frequent in sodium acetate extracts. PMID- 9397558 TI - Austrian Caucasian population data for the quadruplex plus amelogenin: refined mutation rate for HumvWFA31/A. AB - Human identification of biological specimens has undergone immense change since the development of PCR typing systems for forensic casework. In contrast to RFLP and VNTRs, STRs are the method of choice when the investigated genomic DNA is present in low quantity or in degraded shape. In the current study, the X-Y homologous gene Amelogenin has been added to a widely used multiplex PCR amplification system consisting of four tetrameric STR loci (Quadruplex-HumTH01, HumvWFA31/A, HumFES/FPS, and HumF13A1). The modified Quadruplex was used to type 382 unrelated Caucasians from Western Austria. The population data meet Hardy Weinberg and linkage equilibrium expectations, and do not show significant deviations from either US, German, and Turkish Caucasian databases. In an investigation of 382 meioses, two mutations were revealed at the HumvWFA31/A locus. Consequently, the data in this paper provide the conditions for adding Amelogenin to the Quadruplex, and suggest that when doing paternity testing, the mutation rate for the HumvWFA31/A locus must be considered. PMID- 9397559 TI - Validation studies for the genetic typing of the D1S80 locus for implementation into forensic casework. AB - A series of validation experiments were designed to evaluate, according to the Technical Working Group on DNA Analysis Methods (TWGDAM) guidelines, the analysis of the D1S80 locus for casework implementation. Approximately 400 samples from three different populations (Minnesota Caucasian, Minnesota African Americans, and Minnesota Native Americans) were typed to determine allele frequencies. Simulated forensic type specimens (blood, saliva, hair and semen, or vaginal secretions) were typed to demonstrate that deoxyribonucleic acid (DNA) extracted from various tissues of an individual yield the same D1S80 type. Dilution studies were performed and it was determined that a wide range of input DNA (0.5 ng to 40.0 ng) will consistently yield typeable results. The evaluation of DNA from various animals showed that the D1S80 locus is specific to human DNA within the limits of the parameters tested. The reproducibility of the system was tested by duplicate analysis of approximately 200 population samples. Duplicate samples were analyzed on both horizontal and vertical gel systems. In addition, simulated forensic specimens were analyzed by two independent laboratories: the Minnesota Forensic Science Laboratory (MFSL) and the Roche Biomedical Laboratories (RBL). All analyses, including extraction, quantitation, amplification and typing, were performed independently. All typing results for both laboratories were in agreement. By the analysis of mixtures from various simulated casework type mixtures, it was demonstrated that the D1S80 typing system is suitable for analyzing mixtures. In addition to the simulated casework, evidentiary samples from several adjudicated cases previously analyzed by restriction fragment length polymorphism (RFLP) analysis and/or DQA1 were typed at the D1S80 locus. The D1S80 results were consistent with previous RFLP and/or DQA1 results regarding inclusions/exclusions. PMID- 9397560 TI - Analysis of allele distribution for six short tandem repeat loci in the French Canadian population of Quebec. AB - Short tandem repeat (STR) loci represent a rich source of highly polymorphic markers in the human genome which are useful for the purposes of forensic identification and determination of biological relatedness of individuals. Here, as a part of an ongoing extensive study, we report the analysis of a multilocus genotype survey of 642 to 870 chromosomes in the French Canadian Caucasian population of Quebec at six STR loci. The loci HUMCSF1PO, HUMTPOX, HUMTH01, HUMF13A01, HUMFESFPS, and HUMvWA were typed using two multiplex polymerase chain reactions (PCR). Amplified DNA samples were subsequently analyzed by polyacrylamide gel electrophoresis followed by silver staining. The heterozygote frequencies of the loci range from 0.614 to 0.820 (0.661 to 0.818 expected) and the number of alleles from 7 to 12 per locus. Although statistically significant deviation from Hardy-Weinberg expectations of genotype frequencies was noted at some loci by one or more tests, in general, the genotype frequencies are well estimated from the product of allele frequencies at all loci. The most frequent six-locus genotype is expected to occur in the French Canadian population with a frequency of 3.50 by 10(-5) and together, these six loci have an average probability of discrimination of 0.9999985. The study presented here indicates that these six STR loci are informative genetic markers for identity testing purposes in the French Canadian Caucasian population of Quebec. PMID- 9397561 TI - The effects of blood transfusions on PCR DNA typing at the CSF1P0, TP0X, TH01, D1S80, HLA-DQA1, LDLR, GYPA, HBGG, D7S8 and GC loci. AB - Pre-transfusion and post-transfusion blood samples from eight individuals were typed at 10 PCR amplified loci. In no case did the PCR DNA profile of the post transfusion blood sample differ from that of the pre-transfusion profile. PMID- 9397562 TI - Molecular detection of JC virus in embalmed, formalin-fixed, paraffin-embedded brain tissue. AB - Embalmed tissues are adequate for the detection of JC virus in lesions of progressive multifocal leukoencephalopathy (PML) by immunohistologic and molecular methods. JC virus was readily detected in embalmed brain tissue using immunohistochemistry (IHC), in situ hybridization (ISH), and the polymerase chain reaction (PCR). Two brains were removed from bodies that had been embalmed at least 24 h prior to autopsy. They were subsequently post fixed in 10% buffered formalin for 10-14 days before dissection and molecular studies were performed. Though these techniques are not novel, their use in embalmed tissues is. Routine embalming should not eliminate these diagnostic procedures from consideration. PMID- 9397563 TI - GC/MS confirmation of barbiturates in blood and urine. AB - A gas chromatography-mass spectrometric method is described for the quantitative measurement of 6 commonly used barbiturates in blood and urine specimens. The targeted barbiturates are butalbital, amobarbital, pentobarbital, secobarbital, mephobarbital and phenobarbital. They are recovered along with the internal standard, tolybarb, from blood and urine using liquid extraction then alkalated to form the N-ethyl derivatives. The ethylated barbiturates have symmetrical peaks which are well separated from each other on a non-polar methylsilicone capillary column. The derivatives on a non-polar methylsilicone capillary column. The derivatives facilitate quantitations between 50 and 10,000 ng/mL. The day-to day CVs for all 6 barbiturates were between 4 and 9% at 200 and 5000 ng/mL. The method has been extended for identifying other acidic drugs and drug metabolites. They are mainly non-steroidal anti-inflammatory drugs, diuretics, and anticonvulsants. An additional 83 compounds can be qualitatively identified. PMID- 9397564 TI - A false report of product tampering involving a rodent and soft drink can: light microscopy, image analysis and scanning electron microscopy/energy dispersive X ray analysis. AB - The "Pepsi Tamperings" of 1993 resulted in a large number of cases involving foreign objects reportedly found inside canned soft drinks. Although the majority of cases involved medical syringes and metallic objects, one case involved the report of a mouse found inside a can of Caffeine-Free Diet Pepsi. Using light and polarized light microscopy and computer-assisted image analysis, trace evidence and tooth structure from the suspect mouse were matched to scratches and indentions on the suspect can. Scanning electron microscopy and energy dispersive X-ray analysis were used to compare and match particles of gnawed metal from the lid of the suspect can to other particles recovered from the muzzle and stomach of the suspect mouse. The forensic analyses in this case proved the mouse could not have been canned in the soft drink product and refuted the defendant's sworn statements. PMID- 9397565 TI - Estimation of postmortem interval based on colony development time for Anoplolepsis longipes (Hymenoptera: Formicidae). AB - The postmortem interval for a set of human remains discovered inside a metal tool box was estimated using the development time required for a stratiomyid fly (Diptera: Stratiomyidae), Hermetia illucens, in combination with the time required to establish a colony of the ant Anoplolepsis longipes (Hymenoptera: Formicidae) capable of producing alate (winged) reproductives. This analysis resulted in a postmortem interval estimate of 14 + months, with a period of 14-18 months being the most probable time interval. The victim had been missing for approximately 18 months. PMID- 9397566 TI - A killing of Baby Doe. AB - An anencephalic infant died in the Neonatal Intensive Care unit six hours after birth. Eighteen months later, in a discussion of intrusive federal "Baby Doe" regulations with co-workers at the hospital, a registered nurse mentioned that he had found a way to avoid these provisions, and that he had in fact done so on one occasion by killing this particular infant. A co-worker related his story to police, and the "wheels of justice" were set into motion. I describe the chronology of events and the pathologic findings in this case of infanticide, purportedly committed "with mercyaforethought." PMID- 9397567 TI - Tissue distribution of ketamine in a mixed drug fatality. AB - While reports of ketamine abuse are increasing, reports of ketamine deaths and tissue concentrations associated with fatalities are rare. We report here a case of a mixed drug fatality involving ketamine and ethanol. Ketamine analysis was carried out by gas chromatography with a nitrogen-phosphorus detector (NPD). We found the following tissue concentrations: blood 1.8 mg/L; urine 2.0 mg/L; brain 4.3 mg/kg; spleen 6.1 mg/kg; liver 4.9 mg/kg, and kidney 3.6 mg/kg. The blood ethanol concentration was 170 mg/dL. Because an empty nalbuphine ampule was found in the possession of the deceased, the blood was assayed for this opioid compound using a gas chromatography/mass spectrometry (GC/MS) method. None was detected at a limit of detection of 0.02 mg/L. PMID- 9397568 TI - Two traffic fatalities related to the use of difluoroethane. AB - The 18-year-old white male driver and 17-year-old white male passenger of an automobile were killed when their vehicle crossed the median of a 4-lane highway and collided with a minivan. A can of airbrush propellant was found in the automobile of the deceased. The only drug detected during initial toxicological analyses was 130 mg/L ethanol in the blood of the driver. When performing ethanol analysis by headspace gas chromatography, an unidentified peak was observed in the blood of both deceased. This peak was identified as difuoroethane (Freon 152), the propellant in the aerosol can found in the automobile. The concentrations of difluoroethane in the blood of the driver and passenger were 78 mg/L and 35 mg/L, respectively. Based on a literature search we believe that this is the first report of the quantitation of difluoroethane in biological samples. PMID- 9397569 TI - Effects of 3,4-methelenedioxymethamphetamine in decomposing tissues on the development of Parasarcophaga ruficornis (Diptera: Sarcophagidae) and detection of the drug in postmortem blood, liver tissue, larvae and pupae. PMID- 9397570 TI - Findings in gunshot wounds from tandem projectiles. PMID- 9397571 TI - Barbiturates and analgesics in Calliphora vicina larvae. PMID- 9397572 TI - An examination of the binding of aluminum to protein and mineral components of bone and teeth. AB - Aluminum binding by a variety of noncollagenous proteins associated with bone and teeth (osteopontin, osteocalcin and phosphophoryn) and by hydroxyapatite are examined. The proteins bound aluminum with a dissociation constant, KD, of the 10(-7) M or greater at pH = 7. The number of atoms of aluminum bound was found to be related to but not equivalent to the number of phosphorylated serines in osteopontin and phosphophoryn. Osteocalcin bound one aluminum tightly presumably due to its gamma-carboxyl glutamate residues. Hydroxyapatite bound Al3+ tightly releasing 1.5 equivalent Ca2+ per Al3+ bound. Addition of 3 mM Ca2+, close to the total concentration found in animal circulating fluids, did not effect noticeably the amount of Al3+ bound to bone which must have a KD for Al3+ < 10(-6). Silicic acid added after equilibration with all these materials has little effect but neither the proteins nor hydroxyapatite removed much Al3+ from pre-equilibrated Al3+ solution with silicic acid. The results are discussed with regard to Al3+ poisoning. PMID- 9397573 TI - Interaction of iron(II) with bile salts. AB - Iron(II) ions react with small aggregates of cholate, glycocholate, chenodeoxycholate, and deoxycholate to form soluble and colloidal compounds. Taurocholate under conditions used does not react with the Fe2+ ion. Small aggregates of dihydroxy bile salts (predominating in the premicellar region, at concentrations of the bile salt above 1 mmol dm-3) have a larger affinity for Fe2+ compared to those formed from cholate anions. In their interactions with small aggregates of cholate anions, the Fe2+ ion shows an affinity comparable to that of Cu2+ and Cd2+ and somewhat larger than that of Zn2+. Small aggregates of cholate show a higher ability to mask Fe2+ than those of taurocholate and glycocholate. Interaction of glycocholic acid anions with Fe2+ ions is sufficient to prevent iron(II) precipitation. PMID- 9397574 TI - Effects of additional iron-chelators on Fe(2+)-initiated lipid peroxidation: evidence to support the Fe2+ ... Fe3+ complex as the initiator. AB - The addition of chelated Fe2+ ions in a liposomal system often results in a short lag period before peroxidation starts. The addition of a second chelator at the end of the lag period results in an inhibition of the lipid peroxidation. The degree of inhibition depends on the stability constants of the chelator in ligating Fe2+ and/or Fe3+. A more striking inhibitory effect was observed for the chelators with higher stability constant for either or both Fe(2+)- and Fe(3+) complex, but much less inhibition was found for those with lower stability constants for both complexes. Assuming that the "initiator" for iron-dependent lipid peroxidation is formed through the redox process of iron ion and finally emerged at the end of the latent period, the inhibitory effect of the second chelator may be explained as the abstraction of either Fe2+ or Fe3+ from the initiator by an additional free chelator, which results in the decomposition of the initiator. This study supports the hypothesis that a Fe2+ ... Fe3+ complex is responsible for iron-initiated lipid peroxidation. PMID- 9397575 TI - Evidence for a novel heme adduct generated by the in vitro reaction of 2,4,6 trinitrotoluene with human hemoglobin using electrospray ionization mass spectrometry. AB - The bioactivation of nitroaromatic compounds to highly reactive intermediates is responsible for the genotoxic and cytotoxic effects by reaction with DNA and proteins. Due to its continued use as a secondary explosive and its prevalence at contaminated sites, the mechanism of covalent binding of 2,4,6-trinitrotoluene (TNT), or its metabolites, to critical cellular proteins has been of interest. Herein, we report the in vitro reaction of TNT with human hemoglobin under anaerobic and reductive (using sodium hydrosulfite) conditions, yielding a novel adduct between a putative nitrosodinitrotoluene (MW = 211 Da) and the prosthetic heme group (iron protoporphyrin-IX or heme b). While the covalent modification of hemoglobin polypeptide chains by TNT has been established, to our knowledge, this is the first example of a heme-TNT related adduct. This finding could be of relevance in investigation of biotransformation of TNT in subjects exposed to TNT via skin exposure or inhalation. PMID- 9397576 TI - The berenil ligand directs the DNA binding of the cytotoxic drug Pt-berenil. AB - To determine the affinity towards DNA sequences of novel antitumor drugs in comparison with their parental compounds may lead to the design of new analogous drugs with improved antitumor activity. Thus, the affinities of Pt-berenil towards different DNA sites relative to cis-DDP and berenil drugs were analysed using DNase I footprinting and restriction endonuclease analysis. The data show that the Pt-berenil drug inhibits the cutting activity of Hind III enzyme to the same extent as the berenil ligand. In contrast, inhibition by Pt-berenil of the cutting activity of Bam HI enzyme is significantly lower than that of cis-DDP. These results indicate that although the cis-Pt(II) centres of Pt-berenil maintain certain affinity toward G + C regions, which are the main binding sequences of cis-DDP, however, the berenil ligand seems to direct the Pt-berenil molecule towards A + T regions, which are the binding sequences preferred by berenil. In fact, 1H- and 195Pt-NMR spectra of Pt-berenil:nucleoside complexes show that Pt-berenil not only covalently binds to N7 of guanosine but also to N1/N7 of adenosine. PMID- 9397577 TI - Synthesis and antitumor activity of (diamine)platinum(II) complexes of benzylmalonate derivatives. AB - (Diamine)platinum(II) complexes of benzylmalonate derivatives as a leaving group designed in a wide range of lipophilicity and water-solubility were prepared and their antitumor activities were attempted to correlate to their lipophilicity or solubility. A good relationship was observed between their in vitro toxicity and solubility of the title complexes with the same carrier ligand, DACH (trans-(+/-) 1,2-diaminocyclohexane): The most soluble complexes are most cytotoxic whereas the least soluble complexes are least cytotoxic. However, no relationship could be established between their in vivo activity and their lipophilicity or solubility presumably due to other pharmacokinetic factors involved in vivo. The molecular structure of (IPA)2Pt(DBM).2CH3OH (IPA = isopropylamine; DBM = dibenzylmalonate) was determined by X-ray diffraction: space group P2(1)/n, a = 11.433 (3), b = 14.461 (4), c = 17.478 (4) A, beta = 97.25 (3) degrees, z = 4, R = 0.0437. PMID- 9397578 TI - Organotin complexes with pyrrole-2-carboxaldehyde monoacylhydrazones. Synthesis, spectroscopic properties, antimicrobial activity, and genotoxicity. AB - A series of organotin complexes with pyrrole-2-carboxaldehyde 2 hydroxybenzoylhydrazone (H3mfps) and pyrrole-2-carboxaldehyde 2 picolinoylhydrazone (H2mfpp) was investigated. The IR, 1H, and 119Sn nuclear magnetic resonance spectroscopic characterization of all the compounds is reported and discussed in connection with the ligand behaviour of the hydrazone and the structure of the organotin complex. Complexes exhibit antibacterial properties higher than those of the corresponding ligands but they turn out to be less potent than the parent organotin compounds. Sn(H3mfps) (C6H5)2Cl2.2H2O and Sn(Hmfpp)(n-C4H9)2Cl are the most active antibacterial compounds showing MIC values between 3-6 micrograms/ml against Bacillus subtilis and Staphylococcus aureus and between 6-25 micrograms/ml against Escherichia coli; the first compound also strongly inhibits the growth of Aspergillus niger. All the ligands and complexes are devoid of DNA-damaging activity in the Bacillus subtilis rec assay. H2mfpp and its complexes Sn(Hmfpp)(C2H5)2Cl and Sn3(Hmfpp)(mfpp) (C6H5)3Cl6 are shown by the Salmonella-microsome assay to be mutagenic substances in the presence of a metabolic activation system. The obtained results are discussed on the basis of structure-activity relationships. PMID- 9397579 TI - The effects of muscular dystrophy on craniofacial growth in mice: a study of heterochrony and ontogenetic allometry. AB - Mechanical loading of muscles on bones at their sites of attachment can regulate skeletal morphology. The present study examined the effects of muscle degeneration on craniofacial growth, using two strains of muscular dystrophic mice, Mus musculus, differing in pathological severity. We collected radiographic and weight data longitudinally and digitized radiographs to obtain distances between anatomical landmarks in different functional regions of the skull. We then quantified heterochronic and allometric differences among genotypes and between sexes. Because growth is nonlinear with respect to time, we first used the Gompertz model to obtain heterochronic growth parameters, which were then tested with ANOVA. Ontogenetic allometric analyses examined the scaling relationships between various measurements with linear regressions. For most measurements the severely dystrophic mice are significantly smaller in final size than both the control and the mildly dystrophic mice, which are statistically indistinguishable. Measures of total growth and the neurocranium exhibit more differences among groups in heterochronic parameters of early ontogeny because growth in these regions is controlled primarily by brain expansion that ceases early in development. In contrast, the face and mandible exhibit more differences in later growth parameters possibly because of the increased influence of muscles on these regions as growth progresses. The severely dystrophic mice have flatter, more elongate skulls and mandibles than those of the other two genotypes, concurrent with an absence of muscular forces to stimulate growth in a superior inferior direction. PMID- 9397580 TI - The caudal neurosecretory system and its afferent synapses in the goldfish, Carassius auratus: morphology, immunohistochemistry, and fine structure. AB - Morphological features of the goldfish caudal neurosecretory system were investigated by means of immunohistochemical localization of urotensins I and II (UI and UII) and electron microscopic examination of the caudal neurosecretory neurons, the urophysis, and the synaptic neuropil. The aim of the work is to provide a detailed morphological description of the afferent synapses to the caudal neurons and to analyze their distribution through the rostrocaudal extension of the caudal neurosecretory system. Three morphologically different types of neurosecretory cells have been identified according to size and shape: large, medium, and small Dahlgren cells. The three different-sized cells share similar patterns of immunoreactivity with the UI (or oCRF) and the UII antisera. Electron microscopic examination of the synaptic neuropil throughout the caudal system revealed the presence of four types of terminals: dense-cored-vesicle end bulbs (DC), spherical-vesicle end bulbs (S), flattened-vesicle end bulbs (F), and granular-vesicle end bulbs (G). The present study demonstrates that the small Dahlgren cells receive different synaptic inputs from the large and the medium neurosecretory cells. Indeed, G terminals are only found on the small Dahlgren cells, whereas DC, S, and F terminals are distributed on the large, medium, and small Dahlgren cell bodies and proximal processes. PMID- 9397581 TI - Differential distribution of elastic system fibers in control and bronchoconstricted intraparenchymatous airways in the guinea-pig lung. AB - The elastic system fibers were studied at the light microscopic level by using Weigert's resorcin-fuchsin method after oxidation. This study was designed to describe the distribution of these fibers in intrapulmonary guinea-pig airways and to characterize their conformational changes during bronchoconstriction induced by methacholine aerosol. Airways present a palisade of elastic system fibers just beneath the epithelial basement membrane; these fibers are also present in the adventitial connective tissue. Thin fibers link the fibers located in the palisade among themselves and also connect them to those fibers located in the bronchial adventitial tissue, by traversing the airway smooth muscle. During bronchoconstriction, the fibers located beneath the epithelial basement membrane are divided into two components: one follows the epithelial invaginations towards airway lumen, while the other population remains attached through airway smooth muscle to the fibers located in the adventitial connective tissue. At the ultrastructural level, the findings corroborated those of the light microscopy and in addition, disclosed that typical mature elastic fibers and also elaunin fibers attach directly to the basal lamina, a feature that has not been reported previously in other tissues studied. This configuration is compatible with the idea that fibers of the elastic system restrict the mucosal folding during bronchoconstriction, and may also provide energy to restore airway configuration to its normal status after contraction. PMID- 9397582 TI - Sperm ultrastructure of six Australian hylid frogs from two genera (Litoria and Cyclorana): phylogenetic implications. AB - The spermatozoa of four fossorial (Litoria alboguttata, Cyclorana brevipes, Cyclorana novaehollandiae and Cyclorana cryptotis) and two non-fossorial australian hylid frogs (Litoria aurea and Litoria moorei) together with previously examined Litoria (Hylidae: Anura) are compared. In spermatozoal ultrastructure (in particular the structure of the sperm tail) Cyclorana includes species which appear derived (apomorphic) relative to non-fossorial species of Litoria while the fossorial L. alboguttata groups with Cyclorana. All hylid species examined here are united by the bufonoid synapomorphy of a conical subacrosomal cone consisting of separate sheaves and the eubufonoid synapomorphy of a mitochondrial sheath or collar separated by a cytoplasmic canal from the centriolar region and tail. Spermatozoal symplesiomorphies for the Eubufonoidea, seen in Litoria (with the exception of L. alboguttata), are the well developed thin undulating membrane with juxta-axonemal and axial fibre. L. alboguttata, C. novaehollandiae and C. brevipes appear monophyletic in the apomorphic modification of the undulating membrane as a thick, dense structure. In L. alboguttata and C. novaehollandiae this structure retains a swelling, at the free edge, homologized with the axial rod. C. brevipes has a further apomorphy as the undulating membrane forms a parallel-sided dense structure with no separate differentiation of an axial fibre. C. cryptotis, however, retains the plesiomorphic sperm tail with a thin undulating membrane, juxta-axonemal and axial fibre. That these differences in spermatozoal ultrastructure have phylogenetic significance is endorsed by the similarity of the fertilization biology of the species examined. On the basis of sperm ultrastructure three separate lineages are discerned within Cyclorana s. lat.: 1) C. cryptotis; 2) L. alboguttata and C. novaehollandiae; and 3) C. brevipes. The evidence of sperm ultrastructure, supported by previously published molecular, morphological and karyological data, clearly places Litoria alboguttata within the genus Cyclorana. PMID- 9397583 TI - Calcific chronic lateral epicondylitis: a histological and ultrastructural study. AB - Fragments of insertion tissue from right arm common extensor muscle have been collected from a 25-year patient with chronic lateral epicondylitis. Specimens, processed for light (LM) and electron (EM) microscopy, evidentiated a variety of degenerative alterations, such as focal hyalinosis, lipoidosis, collagen fiber redistribution, calcifications and vascular changes. Evidence of collagen normal function maintenance and turnover have been also observed in tenocytes. PMID- 9397584 TI - Interfollicular fibroblasts in the human thyroid gland: recognition of a CD34 positive stromal cell network communicated by gap junctions and terminated by autonomic nerve endings. AB - While epithelial structure and functions have been substantially investigated in many organs, the mesenchymal elements have received less attention. Compared with follicular epithelial cells, there are a few morphological studies on the stroma of human thyroid gland. In order to characterize more fully and assess its possible functions, 15 samples of surgical and autopsy human thyroid tissue were studied by classical histology, immunohistochemistry, transmission electron microscopy, electron microscopic immunohistochemistry, and scanning electron microscopy. In human thyroid gland, the interfollicular connective tissue surrounding the follicles contained collagenous matrix, fibroblasts, unmyelinated nerve fibers with Schwann cells, small blood vessels, lymphatics, lymphocytes, plasma cells, macrophages, and mast cells. At the ultrastructural level, gap junctions between the cytoplasmic processes of interfollicular fibroblasts constituted a novel observation. Immunohistochemistry using a monoclonal antibody against Cx43 confirmed the distribution of gap junctions between stromal fibroblastic cells, which was compatible with the ultrastructural findings. The frequent and intimate association of fibroblastic processes with nerve terminals was also shown. Interfollicular stromal fibroblasts also stained with CD34. The main constituent of the human thyroid stromal tissue was a CD34 positive reticular network involving fibroblasts, mononuclear cells and nerve terminals. It represents a highly ordered stroma, with potential structural and functional similarities to the stroma of bone marrow (Yamazaki and Allen, 1990). PMID- 9397585 TI - The effect of pentoxifylline on ultrastructural picture of type II alveolar epithelial cells and generation of reactive oxygen species during cyclophosphamide-induced lung injury. AB - The aim of the study was to evaluate the effect of pentoxifylline (PTXF) on the production of reactive oxygen species in the rat lungs after intraperitoneal (i.p.) cyclophosphamide (CP) administration (150 mg/kg b.w.) and to draw a correlation between the morphological changes and biochemical findings. Morphological examinations were based on ultrastructural analysis in the transmission electron microscope. It was found that single i.p. CP administration caused destructive changes, particularly within mitochondria and lamellar bodies of type II alveolar epithelial cells (EP II), and within blood vessels of interalveolar septa. The changes were accompanied by an increase in the MDA (malondialdehyde) homogenates. The animals receiving PTXF + CP showed normalization of MDA level and minor damage to EP II confined to alterations in lamellar bodies of EP II. The vascular changes observed after i.p. CP administration were manifested in endothelial swelling and accumulation of neutrophilic granulocytes, monocytes and in the later period blood platelets within the capillaries of interalveolar septa. The cells were tightly attached to endothelium. The changes in the vascular system were accompanied by a significant decrease in the activity of antioxidant enzymes (Cu,Zn-SOD and GSSG-R) in the blood serum collected from the left ventricle of the heart. In the animals given PTXF + CP, a higher decrease was revealed in the activity of these enzymes. Ultrastructural examinations found morphological signs of a facilitated flow, mainly of cellular elements, through the vessels of the interalveolar septa of the lungs. PMID- 9397586 TI - Cell body volume of spinal ganglion neurons: estimation by three different methods. AB - We estimated the mean volumes of two series of nerve cell bodies, one from rabbit and one from rat spinal ganglia by three different methods: a procedure we devised 25 years ago (the circle-fitting method), one of the new stereological methods (the nucleator method) and the method of serial sectioning--the most direct and accurate procedure presently available for estimating cell size. In the case of the rabbit, in which most spinal ganglion neurons have a single nucleolus, the mean volumes estimated by the first two methods are closely similar and deviate by less than 2% from the mean obtained by serial sectioning. In the case of the rat, in which approximately half of the spinal ganglion neurons have more than one nucleolus, the mean volumes estimated by the first two methods are again closely similar, but deviate by about 12% from the mean obtained by serial sectioning. These findings show that: a) both the nucleator method and the circle-fitting procedure are more accurate when applied to neurons with a single nucleolus; b) if certain conditions are respected, not all the methods previously used to estimate cell size give biased results. However, the new stereological procedures are easier and quicker to use than the earlier methods. These findings also show that our previous results obtained by the circle-fitting method are to be considered valid. PMID- 9397587 TI - Cytotoxic effect against HeLa cells of polysaccharides from the lichen Ramalina celastri. AB - The most active polysaccharides which show anti-tumoral activity are (1-->3)-beta D-glucans, branched or not at O-6. Since these structures are sometimes poorly soluble in aqueous media, alpha-D-glucans and their chemical derivatives, which are more soluble, were also studied. The present object is to observe morphological alterations in HeLa cells caused by two different polysaccharides obtained from the lichen Ramalina celastri, which are (1-->3),(1-->4)-linked alpha-D-glucan and its sulphated derivative. The cells were incubated in Eagle's medium in the absence or presence of each polysaccharide and routinely processed and analysed by light and electron microscopy. Even though the alpha-D-glucan altered the cellular volume, cytoplasmic densities, and mitosis, the resulting monolayer was similar to the control. TEM analysis showed cytoplasmic blebbing and the presence of an amorphous electron-dense material free in the cytoplasm and interior membranes. The enhanced injury caused by the sulphated derivative was apparent, altering cell adhesion and causing cell aggregation. Nuclear modifications such as fragmentation and condensation of chromatin under the nuclear envelope, which showed to be convoluted, suggested the occurrence of cell death by apoptosis. PMID- 9397588 TI - Intramural coronary artery disease in swine with naturally occurring hypertrophic cardiomyopathy. AB - Intramural coronary artery disease (ICAD) has been reported in myocardium affected with hypertrophic cardiomyopathy (HCM), but has never been studied in detail with respect to the cell type or lipid infiltration involved in the wall thickening. The lack of heart samples may be one of the rationales to hamper the progress in investigating this disease. Recently, the discovery of naturally occurring HCM in swine has provided an excellent opportunity for the study of ICAD because of the high prevalence of ICAD in this animal. The present study provides a detailed structure feature in the thickened arterial wall of ICAD by both histologic and electron microscopic means. Morphologically, the feature of ICAD is due primarily to the neointimal thickening. Smooth muscle cells (SMC) and extracellular matrix (collagen and elastic fibers) are the major components responsible for the thickened neointima. Fragmentation of the internal elastic membrane is associated with the migration and proliferation of SMC from the media to the intima. Therefore, pigs with HCM may be a potential animal model not only for the study of the mechanism by which SMC migrate and proliferate into intima, but also for the future investigation of interventions in coronary artery occlusion. PMID- 9397589 TI - Gender difference in noise stress-induced ultrastructural changes in rat myocardium. AB - Male and female rats were exposed to noise 6 h daily for 7 days running and the effect of stress was evaluated both on myocardium ultrastructure and plasma corticosterone level. Both sexes showed subcellular alterations of cardiomyocytes; mitochondria, in particular, which resulted the most affected organelles, exhibited diluted matrix and cristolysis; in some areas, the sarcoplasmic reticulum appeared vesiculated. Quantitative analysis of altered mitochondria revealed that atrial myocardium was more affected in males than in females. Moreover, corticosterone plasma assay showed, in exposed animals, a growing increase without significant gender differences. The present findings suggest a more marked involvement of male sex when challenged with noise stressor. PMID- 9397590 TI - A morphometric ultrastructural study of the seminal vesicle of rats submitted to experimental chronic alcoholism. AB - The effects of chronic alcohol ingestion on the secretory epithelium of the seminal vesicle were studied in rats (Rattus norvegicus). Male adult albino Wistar rats were divided into two groups: alcoholic and control. Tips of the seminal vesicle were removed and prepared for light and electron microscopy. Ultrastructural observations on the epithelial cells of the seminal vesicle showed reduced epithelial cell size, decreased apical secretory vacuoles, irregularly shaped nuclei with deep infoldings, increased lipid droplets and dense bodies, a small number of microvilli covering the cell surface, and signs of degeneration. In addition to the hormonal effects, alcohol may act on the secretory epithelium of the seminal vesicle. PMID- 9397591 TI - Exposure to hypercholesterolemic serum modifies the expression of cytoskeletal proteins in cultured endothelia. AB - Arterial endothelial layer dysfunction is considered to be one of the most important events which initiate the development of the atherosclerotic plaque and the cell cytoskeleton plays an essential role in maintaining the integrity of the endothelium exposed continuously to haemodynamic forces. The aim of this work was to study the modifications of the cytoskeletal proteins in the vascular endothelium exposed to atherogenic conditions. A hamster aortic endothelial cell line (HAEC) grown on glass coverslips was exposed for 24 h to hypercholesterolemic or normal homologous serum. Upon staining with Oil Red O and examination by phase contrast and fluorescence microscopy, HAEC incubated with hypercholesterolemic serum appeared heavily loaded with lipid droplets that showed a yellow autofluorescence in UV light and the general aspect of a foam cell. HAEC were incubated with: a) anti-actin serum; b) anti-vinculin monoclonal antibody (MoAb); c) anti-alpha actinin MoAb, and d) anti-talin MoAb, followed by appropriate secondary antibodies coupled with FITC or rhodamine. As compared to normal HAEC, the cells exposed to hypercholesterolemic serum showed a modified pattern for actin and vinculin localization. Actin appeared as a weakly stained network around the nuclear zone whereas vinculin was distributed as small granules throughout the cell cytoplasm. These experimental data suggest that in advanced atherosclerosis, some of the endothelial cytoskeletal proteins undergo modifications which could represent one of the important factors involved in further development of the atheromatous plaque. In addition they indicate that HAEC exposed to hypercholesterolemic serum could represent an in vitro working model for studying the events occurring in the endothelium at advanced stages of atherosclerosis. PMID- 9397592 TI - Structural peculiarities of vascular dendritic cell tubulovesicular system in human atherosclerotic aorta. AB - The principal function of vascular dendritic cells has been suggested to be antigen processing/presentation but how this might occur is not clear. To find out whether the organisation of vascular dendritic cells might allow them to internalise antigens, we used electron microscopy to examine their fine cytoarchitectonics. Tubular and vesicular structures were observed to form continuous nets through which extracellular molecules might gain access to lysosomes. This suggests that vascular dendritic cells might be able to degrade antigens using the same mechanisms as occur in macrophages. We also speculate that antigen processing might be different between type I vascular dendritic cells which possess a hypertrophied tubulovesicular system and type II vascular dendritic cells where the tubulovesicular system is less prominent. The present work indicates that the tubulovesicular system is functionally associated with the extracellular space. This connection with the extracellular space through the system of cisterns and vesicles can allow the transport of different substances entering the cistern nets. PMID- 9397593 TI - Submicroscopic mathematical evaluation of spermatozoa in assisted reproduction. 4. The bovine fertilization (Notulae seminologicae 10). AB - In this paper we apply a modification of the formula of Baccetti et al. (1995) in the evaluation of submicroscopical characteristics of bull spermatozoa used in assisted reproduction. In the present experiment sperm quality is proposed as a useful parameter in predicting the success of fertilization. Our results demonstrate that the percentage of spermatozoa devoid of submicroscopic defects, according to the particular Bayesan formula proposed by us, is clearly correlated with the result of artificial insemination. In fact, the parameter concerning sperm quality obtained in variously successful donors shows a large correlation with fertility power. The synthetic parameters observed are therefore a good tool in the prediction of sperm power in artificial fertilization. The evaluation is mainly concerned with the quality of the acrosomal characters, the status of the chromatin, the shape of mitochondria, the position of the postacrosomal sheath, the perinuclear space and the axonemal pattern. All these characters are expressed with different means in ejaculates. All these data confirm that submicroscopic-mathematical evaluation offers a convincing and reliable diagnosis based upon sperm structure and functions such as acrosomal reaction and cell motility. It has been also demonstrated that sperm quality is a major factor in the success of artificial insemination and it is clearly revealed in the integrity of most of sperm organelles. PMID- 9397594 TI - Microanatomy of the epididymis and vas deferens. AB - In this review, the structural and functional characteristics of the human epididymis, including vas efferents and ductus deferens, are revisited under the morphological and ultrastructural point of view. New surface sperm antigens, enabling the fertilizing power during the epididymal transit, various epididymal microenvironments and their activities on the spermatozoa are reported. This revision is important because the recent procedures of assisted reproduction, aimed at overcoming the cases of obstructive azoospermia, use the spermatozoa aspirated from the proximal segments of the epididymis and suggest a flexibility of the epididymal function with regard to the fertilizing sperm capacity. In my opinion these procedures must be based on an accurate investigation on the function of the different epididymal districts. Moreover, in patients affected by congenital absence of the vas deferens (CAV), the ultrastructural examination of the spermatozoa aspirated from the proximal segments of the epididymis revealed that most of them are defective: in fact the absence of the vas deferens seems to affect the development and the functional properties of the epididymal spermatozoa. PMID- 9397595 TI - Chromium contamination in sediment, vegetation and fish caused by tanneries in the state of Minas Gerais, Brazil. AB - In order to evaluate the chromium contamination from tannery discharges into rivers in the State of Minas Gerais, Brazil, samples of fluvial sediment, vegetation and fish were collected and submitted to chemical analysis. The chromium content in the samples was measured by atomic absorption spectrophotometry. Metal inputs were related to effluent discharges into the rivers. High concentrations of chromium were found in samples when compared with controls. Sediment investigations indicated strong enrichment and high geoaccumulation indices, while chromium concentrations in the analyzed vegetation were higher than those normally found in these materials. Chromium levels in fish exceeded 35 times the Brazilian recommendation value for human intake. PMID- 9397596 TI - Speciation of aluminium in tea infusions studied by size exclusion chromatography with detection by post-column reaction. AB - Aluminium-organic species in tea infusions were studied by size exclusion chromatography, using a Superose 12 HR column, a mobile phase of 0.12 mol l-1 tris(hydroxymethyl)aminomethane adjusted to pH 5.5 and detection by UV-visible spectrophotometry. Aluminium was detected after post-column reaction with pyrocatechol violet at 585 nm and the organic material was detected at 280 nm. Teas of different origin were analyzed; the results indicate that aluminium is bound to the same relatively narrow size-range of large organic molecules in the tea infusions, irrespective of the origin of the tea. PMID- 9397597 TI - Statistical evaluation of ecosystem properties influencing the uptake of As, Cd, Co, Cu, Hg, Mn, Ni, Pb and Zn in seaweed (Eucus vesiculosus) and common mussel (Mytilus edulis). AB - In order to evaluate the influence of geographically varying marine ecosystem properties on the uptake of trace elements in bioindicators, samples were taken of seaweed (Fucus vesiculosus) and common mussel (Mytilus edulis) along the North Sea and Baltic Sea coast. Seasonal variations of the bioindicator status were minimized by sampling within 1 month. Ecosystem properties considered were the geographical position, the salinity and the concentrations of the macroelements Ca, Fe, K, Mg, Na, P and S in the bioindicators. Trace elements studied were As, Cd, Co, Cu, Hg, Mn, Ni, Pb and Zn. Factor analysis of the concentration patterns in the bioindicators and of salinity as a function of location confirmed the influence of the geographically varying salinity on the biological uptake of macroelements and trace elements. This influence of salinity was higher in the case of seaweed than in the case of mussel. Comparison of the geographical courses of the macroelement and trace-element concentrations by cluster analysis revealed corresponding courses for As and Hg in both bioindicators. All other elements showed different courses in seaweed and mussel. Subsequent cluster analysis comparing locations with respect to the macroelement or trace-element concentration patterns in the bioindicators, indicated a clear separation of North and Baltic Sea locations. However, the trace-element concentration patterns provided a regionally less distinctive ecosystem arrangement than those of the macroelement ones. The results of the cluster analysis were verified by discriminant analysis forming groups of locations with respect to geographical position and salinity. Results of discriminant analysis demonstrated, both for seaweed and for mussel as bioindicators, that the location groups formed according to the macroelement concentration patterns corresponded well with the geographical regions in the order of salinity. On the other hand, location groups based on the trace-element concentration patterns again showed a modified less distinctive ecosystem arrangement than the location groups based on macroelement concentration patterns. This confirms modified conditions for the uptake of trace elements in seaweed or mussel in comparison to the uptake of macroelements. PMID- 9397598 TI - Lead levels in roadside soils and vegetation of Damascus city. AB - This study concentrates on seasonal variations of lead levels in roadside soils, vegetables and plants of Damascus city. Lead concentrations in soil samples varied from 78.4 ppm to 832 ppm while mean lead levels in plants ranged from 2.6 ppm to 19.3 ppm; the highest levels were found to be in grass (44 ppm). In addition, lead levels were lower in soil samples during the wet period (December to April) whereas, it is higher in plants during the same period. Moreover, traffic density, distance from traffic roads, climate and area topography (up- and down hill) were influencing factors in lead levels in both the soil and plant samples. The results have also shown that lead concentration in most of the analyzed vegetable samples was high and normal washing does not decrease it to an acceptable level. PMID- 9397599 TI - Heavy metals: leaching from glazed surfaces of tea mugs. AB - Heavy metals (zinc, lead, cadmium, iron, chromium, copper, manganese, nickel and cobalt) were found to leach from the glazed surfaces of tea mugs collected from 13 different pottery units of Khurja (U.P.) and one each from Ghaziabad (U.P.) and Calcutta (West Bengal) determined under different conditions. The leachates used were: tea at 80 degrees C, orange juice at room temperature and 4% acetic acid at room temperature, 40 degrees C and 60 degrees C, respectively. The volume (capacity) of mugs ranged between 200 and 250 ml. The duration for leaching was 24 h in each case without stirring. The concentrations of metals leached in tea at 80 degrees C were found in the range (in microgram/l): Zn, 236-730; Fe, 98 925; Cr, 62-119; Cu, 63-299; Mn, 710-2670; and Ni, 70-80 micrograms/l. The concentrations of metals leached in orange juice at room temperature were in the range (in microgram/l): Zn, 393-1262; Cd, 25-349; Fe, 122-342; Cr, 66-945; Cu, 135-853; Mn, 166-424; and Ni, 70-134 micrograms/l. The concentrations of heavy metals extracted by 4% acetic acid at room temperature were found in the range (in microgram/l): Zn, 18-192; Fe, 143-372; Cu, 51-190; and Mn, 0-48 micrograms/l; at 40 degrees C (in microgram/l): Zn, 118-837; Fe, 124-639; Cu, 230-722; and Mn, 30-63 micrograms/l and at 60 degrees: Zn, 33-900; Fe, 83-576; Cu, 90-685; and Mn, 43-778 micrograms/l, respectively. PMID- 9397600 TI - The impact of age, lactation and dietary habits on PCB in plasma in Swedish women. AB - The mean concentration of the chlorinated biphenyl 2,2',4,4',5,5' hexachlorobiphenyl (CB-153) in plasma from 192 fishermen's wives from the Swedish east coast was on a fresh weight basis 960 (range 80-4300) pg/g and lipid adjusted 160 (range 20-780) ng/g lipid. The concentration of CB-153 in plasma was significantly influenced by age, total lactation time and place of living during childhood and adolescence (fishing village vs. other place). The residential variable probably reflects early life consumption of fish from the Baltic Sea (at the Swedish east coast) contaminated with persistent organochlorine compounds. PMID- 9397601 TI - Isoprene from expired air inside a private car. AB - The concentration of isoprene inside a small-sized parked private car with one person was found to be of the order of 20 micrograms/m3. Isoprene was then the major non-methane volatile hydrocarbon except in strongly traffic-polluted parking places. On driving, with intermediate fan ventilation, the isoprene levels were one order of magnitude lower. In the empty car, the concentrations were still much lower, proving that isoprene originates predominantly from expired air. Air samples were taken on triple-layer adsorbent cartridges and were analysed for volatile hydrocarbons by gas chromatography after thermal desorption. The analytical aluminium oxide column permitted simultaneous determination of a range of reported traffic-emitted hydrocarbons including the carcinogenic 1,3-butadiene and benzene. PMID- 9397602 TI - Hepatitis A outbreak in a socially-contained religious community in rural southern Ontario. PMID- 9397603 TI - Outbreak of cyclosporiasis--northern Virginia-Washington, D.C.-Baltimore, Maryland, Metropolitan area, 1997. PMID- 9397604 TI - The risk and prevention of tuberculosis in travellers. PMID- 9397605 TI - New era of reconstructive microsurgery. AB - Hopfner performed the first successful experimental limb replantation in 1903, and Malt performed the first successful clinical replantation for an above-elbow amputation in 1962. Since then the range of applications for reconstructive microsurgery has expanded rapidly, and it is now widely used for repair of nerves, vessels and lymphatics. In 1972 the first successful case of free flap transfer was performed. Many new applications followed including: (1) coverage of extensive wound defects with exposure of bones, joints, tendons and major vessels which can not be covered with local tissue; (2) vascularized bone transfer for bone defects; (3) vascularized joint transfer for hand joints and the temporomandibular joints; (4) functioning muscle transfer to replace the muscles of the upper limbs and face; (5) toe transfer for missing fingers and thumbs; (6) reconstruction following tumor ablation; (7) reconstruction of congenital anomalies; (8) reconstruction of chest, pharynx and cervical esophagus; (9) transfer of gliding tissue, fascia and sensory flaps for certain injuries. The goals of future reconstructive microsurgery include refinement of procedures, enhancement of functional and aesthetic results, and minimization of the morbidity of donor sites. Improvements of instruments, sutures and computer imaging systems will enable surgeons to perform more accurate reconstructions. Endoscopic harvesting of flaps will be widely used. Surgery will be performed on newborns with certain congenital anomalies. Microsurgery will be increasingly used in conjunction with a prosthesis to improve the function of the prosthesis. Microarthroscopy will be used in operations involving small joints. Advances in microsurgery techniques may also change the results of vascular surgery and tumor resection. PMID- 9397606 TI - Modification of left ventricular isovolumic relaxation time during dobutamine echocardiography as a diagnostic method for ischemic heart disease. AB - BACKGROUND: The diagnostic ability of dobutamine stress two-dimensional echocardiography is limited by the clarity of the echocardiographic image. In this study, left ventricular isovolumic relaxation time (IVRT) was modified and the effect of this modification on the diagnostic accuracy of dobutamine echocardiography (DE) in detecting ischemia was assessed. METHODS: DE was performed in 59 subjects (34 and 25 in the control and patient group, respectively). The results were compared with coronary angiography and stress scintigraphy. We focused on the changes of the IVRT, the corrected IVRT which was the IVRT corrected for heart rate, and the modified IVRT which is the corrected IVRT modified by a new-designed equation. RESULTS: These isovolumic relaxation variables shortened with the increment of dobutamine dosage and were markedly prolonged when ischemia developed. The sensitivity and specificity of DE were 84% and 74%. However, using the prolongation of corrected IVRT and modified IVRT as indicators of ischemia, the specificity increased (from 74% to 91% and to 94%, respectively), without a significant reduction in sensitivity (from 84% to 76% and to 84%, respectively). Patients with positive results for these variables, as compared with negative results, had a significant result of more ischemic segments and a tendency of more diseased vessels. CONCLUSION: The prolongation of these isovolumic relaxation variables during myocardial ischemia improves the diagnostic accuracy of DE and is also well correlated with the severity of ischemia. PMID- 9397607 TI - Malignant thymoma: a review of 44 cases. AB - BACKGROUND: Malignant thymomas are rare neoplasms. Factors affecting prognosis and survival of patients with this neoplasm have been intensively discussed, but the results vary among different studies. To find possible prognostic factors, we designed this retrospective study. METHODS: Forty-four cases of malignant thymomas diagnosed and treated in Chang Gung Memorial Hospital, Kaohsiung, from 1986 to 1996 were reviewed. RESULTS: Of the 44 cases, 24 were male and 20 were female (M:F = 1.2:1). Patient age ranged from 25 to 73 years (median 48 years). Thirty-four cases (77%) belonged to type I malignant thymoma (invasive thymoma) and 10 cases (23%) belonged to type II malignant thymomas (thymic carcinoma). The most frequent histologic type was predominantly epithelial (43%), followed by mixed lymphoepithelial (27%). Six patients had myasthenia gravis. Eleven (25%) patients, including 4 cases of invasive thymoma and 7 cases of thymic carcinoma, showed tumor metastasis to lung, bone, liver, spleen and omentum. The 5-year survival was 73% for patients who underwent total tumor excision and 18% for those who received partial tumor excision or biopsy only. The influence of histologic types on prognosis is not statistically significant (P = 0.434). CONCLUSION: Completeness of tumor excision at initial operation is the most important prognostic factor. Predominantly epithelial and mixed lymphoepithelial types are more aggressive forms with a higher tendency to invasion. PMID- 9397608 TI - Influence of maternal age and weight on second-trimester serum alpha-fetoprotein, total and free beta human chorionic gonadotropin levels. AB - BACKGROUND: The purpose of this study was to assess the relation of maternal age and weight on the maternal serum alpha-fetoprotein (AFP), total human chorionic gonadotropin (hCG) and free beta-hCG levels during the second trimester. METHODS: We collected 419 serum samples from normal singleton pregnancies to assay serum marker levels of AFP, total hCG and free beta-hCG between 14 and 21 weeks of gestation. Maternal age at the day of delivery and maternal weight at the time of sampling were recorded in all cases. The relationship between maternal weight and multiple of the median (MoM) levels of serum markers was analysed by regression models. RESULTS: There was an inverse trend in median MoM levels of serum markers in relation to maternal weight. No significant association between maternal age and serum marker levels was found. CONCLUSION: Because of its impact on serum marker levels, weight correction may be mandatory for further refinement in the maternal serum screening for Down's syndrome. PMID- 9397609 TI - Concurrent 5-fluorouracil, leucovorin, etoposide, cisplatin and radiotherapy for locally advanced non-small cell lung cancer. AB - BACKGROUND: The prognosis of Stage III unresectable non-small cell lung cancer may be improved by concurrent chemoradiotherapy. In this study, we attempted to evaluate the feasibility, tolerance, efficacy and toxicities of the combination of thoracic radiation and chemotherapy with a novel regimen that included 5 fluorouracil, leucovorin, etoposide and cisplatin (the FLEP regimen) in the treatment of this group of patients. PATIENTS AND METHODS: From July 1995 to September 1996, 20 untreated patients with locally advanced non-small cell lung cancer were enrolled in the study. Radiation at a dose of 44 Gy was initially delivered in daily fractions of 2 Gy 5 days a week to the tumor and mediastinum, followed by a boost to the tumor (20 to 26 Gy according to patients tolerance). Concurrently with thoracic irradiation, patients were treated with chemotherapy consisting of cisplatin at the dose of 60 mg/m2/d for 1 day, etoposide at the dose of 60 mg/m2/d for 2 days, and 5-FU 500 mg/m2/d plus leucovorin 50 mg/d infusion for 48 hours. Cycles of chemotherapy were repeated every 3 weeks for a maximum of 3 cycles. RESULTS: Seventeen of 20 patients were assessable. The overall response rate was 70.6% (95% confidence interval = 49-92%). No complete response was achieved. The median response duration for all responding patients is not yet estimable, with a range of 3.5 to 15.5+ months. Eleven patients remain progression-free for 4 to 15 months. The median survival for the entire group is not estimable. The major toxicity was esophagitis. Other grade 3 or 4 toxicities were not frequently observed. CONCLUSION: Combined-modality therapy with FLEP regimen and radiation is a promising treatment with a high response rate and acceptable toxicity for locally advanced non-small cell lung cancer. PMID- 9397610 TI - Monthly measurements of intraocular pressure in normal, ocular hypertensive, and glaucoma male subjects of same age group. AB - BACKGROUND: Recently it has been shown that environmental conditions have a significant influence on intraocular pressure (IOP). Due to differences in inherent constitution, diet and environmental conditions, there is a clear need for well collected IOP data in different countries and ethnic groups. The seasonal variation of IOP has never been described in Pakistani subjects. METHODS: IOP was measured each month over the course of 12 months with the Goldmann applanation tonometer in normal, ocular hypertensive, and glaucoma male subjects. RESULTS: In all groups, the average intraocular pressures in the winter months were highest, while lowest in summer months. The intraocular pressures of spring and autumn months were nearly the same. The intraocular pressure levels in these seasons were between the IOP levels in summer and winter seasons. The difference between highest and lowest IOP was 1.4 +/- 0.2, 3.1 +/- 1.4, and 2.3 +/- 1.1 mmHg, in normal, ocular hypertensive, and glaucoma subjects, respectively. The ups and downs of intraocular pressure were greater in the ocular hypertensive subjects than in the glaucoma patients. CONCLUSIONS: This study confirms that season influences IOP, and concludes that seasonal influence is highest in ocular hypertensive than in normal and glaucoma subjects. As compared to other nations, effect of seasons on IOP seems to be somewhat less pronounced in Pakistan. PMID- 9397611 TI - Application of cryoablation in the management of prostate cancer. AB - BACKGROUND: Radical prostatectomy is the most common and effective therapy for localized prostate cancer. But in addition to its surgical complications, even highly selected series carry a positive margin rate of 35 to 50%. Radiotherapy is another alternative for prostate cancer, but following radiotherapy there have been high positive biopsies reported. Cryosurgery, defined as in situ freezing and hence, devitalization of neoplastic tissues, has currently raised the interest of urologists in the management of localized prostate cancer or failed radiotherapy. MATERIAL: Five patients underwent transperineal cryosurgery of prostate in Chang Gung Memorial Hospital. Among them, three cases were stage D, one stage B and another failed radiotherapy of stage C prostate cancer. All patients received hormone therapy too. RESULTS: PSA declined in 3 patients and biopsies showed intraductal neoplasia. All 5 patients suffered from urine incontinence and one persisted. No mortality has been reported. CONCLUSION: Cryoablation of the prostate is an alternative for treatment of prostate cancer. PMID- 9397612 TI - Prenatal diagnosis of congenital cystic adenomatoid malformation of the lung: four cases report. AB - Congenital cystic adenomatoid malformation of the lung (CCAML) is a rare pulmonary lesion, characterized by excessive overgrowth of the terminal respiratory bronchioles. Prenatal detection and serial sonographic study of fetuses with CCAML can provide information about the natural history of these lesions and reveal most of the nature history of pathophysiologic features which are likely to affect the clinical outcome. This information is crucial to the formulation of a prognosis and a management strategy. We report on four cases of CCAML, three of which involved macrocystic lesions including two cases of type I and one case of type II. Only one microcystic lesion, a type III CCAML, was identified in these patients. All of the cases were diagnosed by ultrasound between the 21 and 24 weeks of gestation. Fetal hydropic change was noted in all four cases. All of the parents opted for termination of pregnancy before fetal viability. Post-mortem examination confirmed the diagnosis in all four cases. PMID- 9397613 TI - Sustained complete remission after incomplete chemoradiation complicated with pelvic cellulitis in a patient with recurrent cervical carcinoma. AB - We report on a 52-year-old female patient with a bulky, recurrent cervical carcinoma involving the vagina and bladder, who developed entero-recto vesicovaginal fistulas and sepsis with pelvic cellulitis after external radiation of 40 Gy and 2 courses of concurrent chemotherapy. Chemoradiation was interrupted and an ileostomy was performed. After recovery, no residual tumor was detectable. Thirteen months after ceasation of chemoradiation, repair of vesicovaginal and rectovaginal fistulas via posterior sagittal approach was performed. Revision of double bowel ileostomy and ileo-T-colostomy was performed 17 months later. The patient enjoyed the restoration of enteral and urinary function only temporarily. She developed rectovesical fistula and underwent an ileostomy again 6 months later. She had another episode of peritonitis and upper gastrointestinal bleeding and expired at 4 years from initiation of salvage therapy. She had no evidence of cancer recurrence during a series of laparotomies and biopsies. The dramatic regression of the tumor may be attributed to its extraordinary radiosensitivity or chemosensitivity. The acute pelvic inflammatory complications may also contribute to the tumor cell killing. The prognosis of recurrent cervical carcinoma is invariably poor except in small tumors confined to vagina. This case gives support to the efficacy of chemoradiation and the potential role of biologic therapy in treatment of this dismal disease. PMID- 9397614 TI - Salmonella typhimurium brain abscess in a six-month-old infant: a case report and review of the literature. AB - We report the case of a six-month-old male infant with brain abscess caused by Salmonella typhimurium. Upon admission, he was suffering from fever, diarrhea, drowsiness and convulsion. Salmonella meningitis was identified by CSF examination. Following failure of antibiotic therapy to control his fever, brain computerized tomography (CT) was ordered 5 days later and revealed a brain abscess. He received surgical excision of the abscess and recovered completely after receiving ceftriaxone therapy for 8 weeks. The case of our patient, together with 11 cases of Salmonella brain abscess from the English literature are reviewed. There was a male preponderance among these patients (male: female = 2.67 : 1) and the majority were less than one year old. Salmonella typhimurium, typhi, and enteritidis occurred most frequently. Fever, seizure, signs and symptoms of increased intracranial pressure and change in mental status were the most common clinical features. Purulent meningitis was a major predisposing factor. Successful treatment was associated with early identification, prompt surgical intervention, high dose, long-term antibiotic therapy, and close follow up for possible recurrence and to determine the presence of neurological sequelae. PMID- 9397615 TI - Internal iliac artery aneurysmo-rectal fistula associated with multiple aortoiliac aneurysms. AB - Fistular communication between an internal iliac artery aneurysm and rectum presenting as massive lower gastrointestinal tract bleeding is a rare entity in clinical practice. Prompt diagnosis and experienced therapeutic application determine the outcome. Herein we report the successful management of such a complication. A 68-year-old male had multiple aneurysms over the abdominal aorta and bilateral iliac arteries. It was the largest aneurysm arising from the right internal iliac artery which ruptured into the rectum and resulted in massive hematochezia. After extraanatomical bypass with right axillo-femoral and femoro femoral crossover grafts to restore the circulation to the bilateral lower limbs, the infrarenal abdominal aorta just immediately above the proximal aneurysm was transected and closed as a blind stump. All the aneurysms were included in this resection and as much of the infected aneurysm tissue was debrided as possible. The rectum was exteriorized using Hartmann's procedure. The patient survived the operation and was discharged in good condition. PMID- 9397616 TI - Cyclopia in one of discordant twins: a case report. AB - Cyclopia is an uncommon congenital anomaly resulting from arrest of the development of the anterior end of the neural plate. It is always associated with abnormalities of the brain. Cyclopia has never been reported in one twin only. In this report, we describe a case of cyclopia in a female infant with normal karyotype who was one of discordant twins. The infant died perinatally. The parents were healthy and not sanguineously related. Prenatal imaging studies revealed cyclopia, holoprosencephaly and hypognathia. Postmortem examinations revealed one orbit, one eyeball and absence of the nose. The central nervous system included rudimentary cerebral hemispheres with fusion of the two lateral ventricles and the third ventricle forming a single large cavity. The superior aspect of each cerebral hemisphere consisted of only a transparent membrane. The olfactory bulb nad tract was absent. The optic nerves were not identified. PMID- 9397617 TI - Cardiac perforation and tamponade induced by external cardiac massage: a case report. AB - A 76-year-old woman suffered from sudden loss of consciousness while sitting in a chair. She was sent to a local hospital and found to be in shock. After a brief period of external cardiac massage, she was transferred to our hospital. In our emergency department she was lethargic with cool, clammy extremities. Her blood pressure dropped from 113/53 mmHg on arrival to 72/42 mmHg 2 hours later. Echocardiography showed massive pericardial effusion, fair left ventricular contractility and no abnormal segmental motion. The echocardiographic appearance suggested fibrin-like substance in the pericardial space, which was felt to indicate the presence of blood. Enhanced chest computerized tomography showed extravasation of contrast medium from the right ventricular outflow tract. At surgery, a small perforation was found at the infundibular area of the right ventricle, and a total of 500 mL of blood had accumulated in the pericardial space. She was discharged 7 days postoperatively, having made an uneventful recovery. External cardiac massage may cause cardiac disruption, and this should be considered in patients who have secondary hemodynamic instability following successful cardiopulmonary resuscitation. PMID- 9397618 TI - Total knee arthroplasty in a rheumatoid arthritic knee with large geode: a case report. AB - Geodes (subchondral cysts) are a well-known manifestation of rheumatoid arthritis. Solitary cysts or cysts larger than 2 cm are not generally found in the knee joint of patients with rheumatoid arthritis (RA). We report a case of RA involving both knees with a giant geode over the right proximal tibia. Surgical treatment was performed including synovectomy, cyst enucleation and packing of autogenous bone chips followed by primary total knee arthroplasty. The postsurgical result was excellent with the knee restored to good function and complete healing of the cystic lesion. PMID- 9397619 TI - Fatal Haemophilus influenzae pneumonia: two cases report. AB - Haemophilus influenzae is a common cause of acute childhood pneumonia. Most Haemophilus pneumonia generally follow a benign course with occasional complications of pleural effusion, pneumothorax or pneumatocele. Deaths following invasive Haemophilus pneumonia have rarely been reported in children older than 3 years of age. We report 2 deaths in children presenting with fulminant pneumonia, complicated by sepsis and adult respiratory distress syndrome despite vigorous antibiotic therapy and full resuscitative measures. PMID- 9397620 TI - Antibiotic prophylaxis and surgery. PMID- 9397621 TI - Attic cholesteatoma and extensive tympanosclerosis. PMID- 9397622 TI - Vocal fold scar. PMID- 9397623 TI - Oral-facial-digital syndrome type I. PMID- 9397624 TI - Mucocele of the pterygoid recess of the sphenoid sinus: endoscopic microdebrider excision via a transnasal sphenoidotomy approach. PMID- 9397625 TI - Practice guidelines and liability implications. AB - It should be clear that the authors of this article are not enamored of practice guidelines, thinking them of much more potential harm than good. On the other hand, if practice guidelines are deemed essential, then they must be written with the goal of quality patient care advocacy and with an eye toward the possible damage they may do. PMID- 9397626 TI - Wound infection in head and neck surgery: implications for perioperative antibiotic treatment. AB - Perioperative antibiotic treatment significantly reduces the risk of postoperative wound infection and is cost-effective in clean-contaminated head and neck operations. A clear consensus on the most suitable single agent or combination is, however, lacking. Most surgical wound infections involve both gram-positive and gram-negative aerobes and anaerobes; some organisms may exhibit antibiotic resistance through beta-lactamase production. Comparative trials have indicated that combinations with both aerobic and anaerobic activity provide protection superior to that achieved with single agents active against only aerobic pathogens. Recent results suggest that the beta-lactam/beta-lactamase inhibitor combination ampicillin/sulbactam is cost-effective for perioperative treatment of patients undergoing head and neck surgery. PMID- 9397627 TI - Inner ear immunology and allergy: an overview of current day concepts. PMID- 9397628 TI - p53 immunostaining of surgical margins as a predictor of local recurrence in squamous cell carcinoma of the oral cavity and oropharynx. AB - Local and regional recurrence is the principal reason for treatment failure in squamous cell carcinoma (SCC) of the head and neck. The conventional method of evaluating surgical margins for cellular atypia does not always predict risk of local recurrence accurately. Immunostaining of surgical margins for tumor markers may provide a more precise evaluation of risk of local recurrence. Paraffin embedded tissue blocks of surgical margins from 24 patients with oral cavity and oropharyngeal squamous cell carcinoma were immunostained for p53 protein. Fifty eight percent of the patients had at least one margin stain positive for p53, including eight of ten patients whose SCC recurred locally. The sample odds ratio test predicted a 5.333 times higher chance of local recurrence with at least one p53 positive surgical margin. The implications of these results for patient management and further investigations will be discussed. PMID- 9397629 TI - Mucocele of the sphenoid sinus due to an osteoma. PMID- 9397630 TI - Osteoma of the base of the tongue. AB - A 14-year-old girl developed a firm mass at the base of the tongue. Computed tomography indicated marked density suggesting either a foreign body or bony tissue. A thyroid scan confirmed the presence of a normally sized and positioned gland. The mass was removed in toto and found to be an osteoma. This is the first report of a case in which the diagnosis of this rare developmental lesion of the tongue was achieved preoperatively based on clinical and radiologic information. This experience should lead to greater awareness of this entity in the future. Recognition of this entity in the pediatric age group is especially useful in avoiding misdiagnosis of other, potentially more aggressive types of tongue mass lesions. Our case demonstrates that it is possible to detect this entity using computed tomography. The dense calcification is truly characteristic of the tumor and may be relied upon to exclude alternative soft tissue mass lesions. While other forms of osseous and cartilagenous neoplasms, such as extraskeletal osteosarcoma and chondrosarcoma, have been reported arising in the tongue, their malignant nature should otherwise be readily apparent. Osteoma of the tongue is the favored diagnosis when mature bone tissue is imaged at the base of the tongue. PMID- 9397631 TI - Development and optimization of a purification strategy for the venom of the scorpion Parabuthus transvaalicus. AB - In search for the toxic components constituting the venom of the southern African scorpion Parabuthus transvaalicus we compared and optimized several chromatographic separation protocols resulting in an efficient venom purification strategy. We found that the sequential combination of gelfiltration (Superdex) together with reversed phase chromatography (C2/C18)-but not cation exchange (Mono S)-stood surety for the best purification. PMID- 9397632 TI - Patient satisfaction with nurse practitioner care in primary care. PMID- 9397633 TI - Are you linking competence to outcomes? PMID- 9397634 TI - What do you call your patients? PMID- 9397635 TI - Four steps to creating quality indicators across sites. AB - As multi-hospital systems develop, the demand to compare performance across sites has become an organizational imperative. This article describes the process a six hospital system used to create clinically reliable and valid quality assessment indicators. PMID- 9397636 TI - Functional teams lead to Joint Commission success. AB - As the Joint Commission on Accreditation of Healthcare Organizations (Joint Commission) has shifted the focus of its survey process to the organization as a whole, new techniques are needed to prepare for surveys. Staten Island University Hospital used teams based on the eleven important functions to achieve a successful survey. PMID- 9397638 TI - Does employee involvement work? Yes, sometimes. AB - Employee involvement per se is not always effective for improving performance and/or employee attitudes. Rather, there are several different forms of employee involvement, some of which are effective, while others are not. This article describes seven forms of employee involvement, giving examples, and summarizes research findings for each form, concluding with a summary of which are the best and which are worst. This article also describes what is necessary for effective employee involvement, focusing on management commitment and training for both management and employees. PMID- 9397637 TI - Quality plan for a product line. AB - Continuous Quality improvement (CQI) has undergone radical change as health care facilities merge, expand, and modify their existing services. CQI has shifted from a centralized position in health care organizations, to unit based, to product lines. This paper describes one product line's endeavors to develop a Quality Plan to direct CQI activities. One particular strength of our innovation is that the Quality Plan was developed with attention to the important balance of interdisciplinary cooperation and maintenance of appropriate discipline boundaries. PMID- 9397639 TI - The future of the quality professional in health care. AB - Radical changes in health care, organizations, and the quality science have converged, radically altering the role of many, including the quality professional. The very existence of this role remains in question. Should it continue, radical changes are in store for its conduct. It is critical that quality professionals today remain aware of the issues, and strategically move forward to chart a quality direction for both their organizations and their own careers. PMID- 9397640 TI - Verifying nursing home care quality using minimum data set quality indicators and other quality measures. AB - Researchers, providers and government agencies have devoted time and resources to the development of a set of Quality Indicators derived from Minimum Data Set (MDS) data. Little effort has been directed toward verifying that Quality Indicators derived from MDS data accurately measure nursing home quality. Researchers at the University of Missouri-Columbia have independently verified the accuracy of QI derived from MDS data using four different methods; 1) structured participative observation, 2) QI Observation Scoring Instrument, 3) Independent Observable Indicators of Quality Instrument, and 4) survey citations. Our team was able to determine that QIs derived from MDS data did differentiate nursing homes of good quality from those of poorer quality. PMID- 9397641 TI - Conducting unit-based research to improve quality of care. AB - How does a tertiary care medical center without a primary academic nursing affiliation improve quality of patient care by conducting unit-based nursing research? The organization's strengths and opportunities include top management support, an infrastructure for development of nurse researchers, a specific patient care coordinator role, research supporting organizational decision making, and success in obtaining nursing research grants. The effects of limited consultative support, statistical avoidance, reorganization, competing staff demands, and lack of funding must be minimized. Strategies include: continue communication with top management, provide meaningful development opportunities and technical research support, build on completed research, use multidisciplinary teams, and change thinking to maximize the research opportunities that already exist. PMID- 9397642 TI - Anatomy of an OPTI dissected: structure, function, and the impact of budget reconciliation legislation. PMID- 9397643 TI - A snapshot of osteopathic medical education in 1997. PMID- 9397644 TI - Forecast for osteopathic medical education programs in the for-profit hospital environs. PMID- 9397645 TI - Examining quality improvement vs cost containment. PMID- 9397646 TI - History of osteopathic medical education accreditation. PMID- 9397647 TI - Undergraduate osteopathic medical education. PMID- 9397648 TI - Income and expenditures of osteopathic medical colleges. PMID- 9397649 TI - Osteopathic graduate medical education. PMID- 9397650 TI - Certification of osteopathic physicians. PMID- 9397651 TI - AOA continuing medical education. PMID- 9397652 TI - Research programs of the AOA and their role in osteopathic medical education. PMID- 9397653 TI - The anatomy of an OPTI: Part 2. The CORE system. Ohio Osteopathic Hospital Association. Ohio Association of Osteopathic Medical Directors. Ohio Osteopathic Association. AB - In July 1995, the American Osteopathic Association (AOA) Board of Trustees passed new regulations regarding the accreditation of osteopathic graduate medical education (GME) by establishing the Osteopathic Postdoctoral Training Institutions (OPTI) system. This system must be phased in by July 1999. The principal changes resulting from the OPTI system include establishing requirements for college cosponsorship of GME programs and for the number of residency programs, interns, and residents to be trained by the OPTI. In essence, OPTI is an osteopathic acronym for consortium. Each OPTI must include at least one college of osteopathic medicine (COM) and one AOA-accredited hospital. The OPTIs will be subject to interval AOA inspections and will be required to demonstrate a governing system, mission statement, organizational structure, and the presence of faculty development programs. The first article in this two-part series, published in the October JAOA, provided a general blueprint for OPTI building and presented both positive and negative issues germane to the formation of OPTIs. Part 2 reinforces the considerations outlined in Part 1 by describing the formation of a large OPTI--the Ohio University College of Osteopathic Medicine (OU-COM) Centers of Osteopathic Regional Education (CORE) system. Key features are described, including the mission statement, organizational structure, committee system, governance, GME programs, operations, and budget. PMID- 9397654 TI - Pathologic and diagnostic considerations in onychomycosis. AB - Understanding the physiology and function of the nail unit and its potential avenues of invasion, and properly identifying invading organisms are two key aspects of using the newer therapies available for the treatment of onychomycosis. This article discusses the most common pathologies of onychomycosis, as classified by the sites of entry of the invading fungi. Susceptibility factors leading to infection are also discussed. Obtaining proper tissue samples, using appropriate tests and culture media, and accurately interpreting test results are all paramount to correct identification of the invading organism and, in turn, to effective prescribing. When fungal-growth results do not support the clinical symptoms, or if a more specific identification of the organism is required, additional diagnostic tests are available and are outlined here. PMID- 9397655 TI - Impact of onychomycosis on quality of life. AB - The author reviews the current knowledge of the impact of onychomycosis on quality of life. Like other visible disorders of the integumentary system, onychomycosis affects many aspects of life, including physical functioning, interpersonal interactions, and emotional state. After examining the nature of quality of life and the study instruments used to measure it, the author reviews several studies that have examined the relationship between onychomycosis and quality of life. The author concludes that onychomycosis is a significant medical problem that has a great impact on patients' lives and should therefore be treated as definitively as possible. PMID- 9397656 TI - Onychomycosis. Baseline results of an observational study. AB - The investigators present an analysis of baseline quality-of-life and patient management approaches from an observational study of 150 patients being treated by podiatric physicians and dermatologists for onychomycosis. The majority (73%) made the initial office visit specifically because of their onychomycosis. Both men and women indicated that they had substantial physical discomfort as well as concerns related to appearance. Women reported significantly more problems than did men as a result of their onychomycosis. Physicians reported that 54% of patients suffered from toenail discomfort, 36% had pain while walking, 40% reported that their condition limited wearing of shoes, and 67% were embarrassed by the condition. The results of this study suggest that the treatment approach of podiatric physicians is more likely to address the palliative concerns of patients with onychomycosis, while the approach of dermatologists is more likely to attempt a definitive cure. PMID- 9397657 TI - Oral treatment options for onychomycosis. AB - The author discusses the new oral antifungal agents for the treatment of onychomycosis. The history, mechanisms of action, efficacies, dosing, safety profiles, and costs of itraconazole, terbinafine, and fluconazole are reviewed. The author emphasizes that use of these effective antifungals represents an important paradigm shift for podiatric physicians away from the palliative therapy of nail debridement to a potentially curative treatment. PMID- 9397658 TI - Practice management and managed-care issues in onychomycosis. AB - The author gives a general overview of recommendations for practice success in a managed-care environment and describes a new practice paradigm for the treatment of onychomycosis. Recommendations are provided for ensuring optimal reimbursement, particularly in the treatment of managed-care and Medicare patients. PMID- 9397659 TI - ECG of the month. Questions. Ventricular tachycardia. PMID- 9397660 TI - Comparison of conventional and deep plane facelift. AB - Revitalization of the aging face is a complex process that must include several components all working together in harmony to create a natural, youthful appearance. As more details of the facial anatomy have been described, the facelift operation has expanded tremendously to include deeper layers of the face with a goal of achieving both a longer lasting effect from surgery and a more complete recontouring effect of the melolabial fold. A comprehensive understanding of the anatomy of the face and the different procedures available is necessary in order to perform facelift surgery effectively and safely. PMID- 9397661 TI - Radiology case of the month. Delayed onset of muscular soreness in the right hip. Delayed onset muscle soreness (DOMS). PMID- 9397662 TI - The journal 100 & 150 years ago. New Orleans Medical and Surgical Journal. November 1847 & 1897. PMID- 9397663 TI - Lafayette Community Health Care Clinic and the Lafayette Parish Medical Society Alliance ... a working partnership. PMID- 9397664 TI - Nicotine addiction and smoking cessation. AB - Cigarette smoking remains the greatest threat to public health in our society and is responsible for one of every five deaths in the United States. While the prevalence of smoking has declined dramatically over the past 30 years, over 26% of Americans continue to smoke. In Louisiana, one third of adult men and one quarter of adult women smoke. The irony of this problem is that smoking is a modifiable behavior. To better counsel their patients regarding smoking cessation, health care practitioners should be aware of the health risks of smoking, the benefits of smoking cessation, and therapies available to assist their patients in breaking the habit. This article reviews the current literature and addresses these issues. PMID- 9397665 TI - Heritable arrhythmias, cardiomyopathies, and valvulopathies. AB - Arrhythmias, cardiomyopathies, and valvulopathies are the most frequent forms of heart disease that occur during the years of peak productivity. They have interrelated pathogenetic bases and may occur in combinations. Cases in which there is involvement of other body systems are so frequent as to suggest a need for careful cardiological evaluation of any patient with dysmorphic features or an inborn error of metabolism. Recent advances in embryology and molecular genetics have increased our understanding of the heritable arrhythmias, cardiomyopathies, and valvulopathies but clinical delineation of all of them predated those developments. Symptomatology may be confusing or absent, so electrocardiogram and echocardiogram are required for diagnosis. Holter monitoring is productive if echocardiogram and electrocardiogram are not diagnostic. For some varieties of heritable arrhythmias, cardiomyopathies, and valvulopathies, other diagnostic modalities are required. Differentiation between heritable and non-heritable varieties may require testing of relatives. Members of the immediate family are most likely to be affected. For X-linked and recessive types, more extensive genealogical investigation may be required. Medical management, prognosis, and genetic counseling are highly dependent on thorough elucidation. PMID- 9397666 TI - Treatment evaluation of a patient with Hodgkin's lymphoma: a case report. PMID- 9397667 TI - A pregnant woman with lupus Tulane Renal Biopsy Conference. PMID- 9397668 TI - Managed care tort liability. PMID- 9397669 TI - The art of medicine. How to get the most out of hospice care. PMID- 9397670 TI - Making the hospice decision. PMID- 9397671 TI - Michael H. Dawson, MD. Injury helps physician appreciate patient's pain. PMID- 9397672 TI - But it's not the money. The real reasons why membership is so important. PMID- 9397673 TI - Heads up--here's what's coming at you in economics and legislation. PMID- 9397674 TI - James L. McGauley, MD. Getting it all together on patient information. PMID- 9397675 TI - Truths and myths. Narcotic medication usage for pain management. PMID- 9397676 TI - The spontaneous ultraweak luminescence of living systems. AB - The results of experimental research on ultraweak photon emission carried out at the Institute of Physics, Catania University, are presented here. A correlation has been demonstrated to exist between photon emission and growth activity in the germination of soya seeds and also during the exponential phase of the growth of yeast cell cultures. In addition an experiment on the self irradiation of yeast cells is reported that confirms the mitogenic effect hypothesized by Gurwitsch. PMID- 9397677 TI - Oncogenous micro-organisms are confined to Mycoplasmas and Helicobacter pylori. AB - Among the various micro-organisms isolated from man's cancerous tissues including the blood of such patients several species were described as new to science. Their identity is being discussed and the conclusion made that none of the scientific names proposed as new for these micro-organisms can be utilized taxonomically. Aberrant red cells have been commonly confounded with some supposed blood parasites. Current research of micro-organisms suspected of playing a role in oncogenesis is confined to two categories of true or opportunistic pathogens: Mycoplasmas with respect to unspecified cancers and Helicobacter pylori in regard to stomach neoplasms. PMID- 9397678 TI - EH: a novel protein-protein interaction domain potentially involved in intracellular sorting. PMID- 9397679 TI - A CTD function linking transcription to splicing. AB - Since its discovery in 1985, the function of the C-terminal domain (CTD) of RNA polymerase II has been a puzzle. Recent studies suggest that the CTD functions as a linear platform for assembly of complexes that splice, cleave and polyadenylate pre-mRNA. A new set of CTD-associated SR-like proteins (CASPs) have been implicated in pre-mRNA processing and transcription elongation as a component of the emerging 'transcriptosome'. PMID- 9397680 TI - A putative nucleic acid-binding domain in Bloom's and Werner's syndrome helicases. PMID- 9397681 TI - A eukaryotic XPB/ERCC3-like helicase in Mycobacterium leprae? PMID- 9397682 TI - ATP synthase: an electrochemical transducer with rotatory mechanics. AB - ATP synthase (F0F1-ATPase) uses proton- or sodium-motive force to produce ATP form ADP and P(i). Three lines of experiment have recently demonstrated large scale intersubunit rotation during ATP hydrolysis by F1. We discuss how ion flow through the membrane-intrinsic portion, F0, may generate torque and how this might be transmitted between stator and rotor to finally expel spontaneously formed ATP from F1 into water. PMID- 9397683 TI - The DNA polymerase alpha-primase complex couples DNA replication, cell-cycle progression and DNA-damage response. AB - The highly conserved DNA polymerase alpha-primase complex (pol-prism) is the only eukaryotic DNA polymerase that can initiate DNA synthesis de novo. It is required both for the initiation of DNA replication at chromosomal origins and for the discontinuous synthesis of Okazaki fragments on the lagging strand of the replication fork. The dual role of pol-prim makes it a likely target for mechanisms that control cell-cycle S-phase entry and progression. PMID- 9397684 TI - Inositol lipid 5-phosphatases--traffic signals and signal traffic. AB - Several novel phosphoinositide 5-phosphatases have been identified and characterised, revealing a growing family of regulators of inositol lipid dependent processes. The features of these proteins, their likely partners and their involvement in signal transduction and membrane traffic is discussed. PMID- 9397685 TI - Dynamic mutation: possible mechanisms and significance in human disease. AB - Increases in repeat-DNA copy number are the molecular basis of a growing list of human genetic diseases, including fragile X syndrome, myotonic dystrophy, Huntington disease and a form of epilepsy. Repeat-DNA sequences undergo a unique process of dynamic mutation, the common properties of which probably reflect common molecular events. This form of mutation is no longer restricted to trinucleotide repeats, because repeats of different length have been found to undergo expansion. PMID- 9397686 TI - The Maf transcription factors: regulators of differentiation. AB - Since the identification of the v-maf oncogene in an avian tumor virus, the Maf protein family has grown rapidly, forming a unique subclass of basic-leucine zipper transcription (bZIP) factors. Maf family members appear to play important roles in the regulation of differentiation. PMID- 9397687 TI - Linking databases and organisms: GenomeNet resources in Japan. PMID- 9397688 TI - The protein kinase resource. PMID- 9397689 TI - Tritium-labeled thymidine and early insights into DNA replication and chromosome structure. PMID- 9397690 TI - Certified occupational health nursing. Job analysis in the United States. AB - Specialty nursing certification programs, such as that administered by the American Board for Occupational Health Nurses, Inc. (ABOHN), must be firmly based on current practice to maintain validity. To determine this, ABOHN performed its most recent job analysis and role delineation study between 1992 and 1994. A comprehensive survey tool was developed by ABOHN Board members, and administered to all 3,805 certified occupational health nurses in practice at the time of the study. With a final return rate of 42.7%, the results were believed to be representative of the knowledge, skills, and abilities needed to practice occupational health nursing in the United States at the proficient level of practice. The results of the study formed the basis for the ABOHN test blueprints and the creation of two credentials for occupational health nurses: the Certified Occupational Health Nurse (COHN) and the Certified Occupational Health Nurse Specialist (COHN-S). PMID- 9397691 TI - Varicella vaccination. An overview for the occupational health nurse. AB - 1. Varicella-zoster virus is highly communicable. Non-immune adults are susceptible to infection and, if infected, have increased risk of complications. 2. Adult varicella infection poses significant economic loss for the infected employee and potentially for exposed coworkers and the employer. Health care employers also must be concerned about transmission of varicella infection to clients. 3. Varicella vaccine is now available, well tolerated, and prevents infection in approximately 94% of vaccinees. The occupational health nurse needs to consider varicella vaccination for employees susceptible to infection. PMID- 9397692 TI - Psychological effects of stress from restructuring and reorganization. Assessment, intervention, and prevention strategies. AB - 1. Demands from the economy, market competition, and the political arena have led to restructuring, reorganization, and change in the workplace. 2. Work behaviors indicative of stress are costly to organizations. 3. Strategies for intervention and prevention should be directed at both managers and workers and can result in cost savings for the organizations. 4. A collaborative approach in which occupational health nurses work with other services inside the organization, and with community organizations and services outside the organization, is important. PMID- 9397693 TI - Project management for occupational health nurses. PMID- 9397694 TI - Workers' compensation litigation review: Part I. PMID- 9397695 TI - AAOHN and ACOEM consensus statement for confidentiality of employee health information. PMID- 9397696 TI - Team building. A powerful learning organization approach. PMID- 9397697 TI - Managing staff transitions. Change is never easy. PMID- 9397698 TI - The integrated evaluation of effectiveness and efficiency. PMID- 9397700 TI - Impact of managed care decisions on cancer care. PMID- 9397699 TI - Celebrating our triumphs. Moving forward with hope. PMID- 9397701 TI - The value of a well-placed stoma. AB - PURPOSE: The author describes the purpose, procedure, and practice of stoma-site marking. Patient anxiety and other emotional implications of ostomy surgery are explored, and expenses related to supplies and reimbursement issues are discussed. OVERVIEW: Patients with specific advanced colorectal, bladder, ovarian, or endometrial cancers sometimes require surgery that controls malignancies but necessitates ostomy creation. Despite a well-placed and well constructed stoma, adjustment to an ostomy is difficult and lengthy. Maladaptive behaviors are exacerbated when an ostomy is constructed poorly and positioned poorly on the abdominal wall. Additional expenses are incurred when this stoma requires a customized and complex pouching system. CLINICAL IMPLICATIONS: Malformed ostomies, leaking ostomy pouches, or skin irritations impact negatively on the patient and family members and may result in treatment delays. The patient's body structure, fat stores, occupation, and clothing style must be considered when determining the appropriate stoma site. Attention also must be paid to psychosocial support so that both emotional and physical healing occur. PMID- 9397702 TI - Women's perception. Breast cancer treatment and sexuality. AB - PURPOSE: The authors describe women's perceptions of how their sexuality was affected by treatment for breast cancer. This description is part of a larger study that tested the psychometric properties of a sexuality measure. DESCRIPTION OF STUDY: This exploratory study analyzed 105 women's responses to one question that asked them to describe how their sexuality had changed since their breast cancer diagnosis. Two researchers independently used content analytic procedures to identify themes that emerged from the data. RESULTS: Four themes emerged: Physical Sexual Functioning, Relationship Quality, Psychological Self, and Self as Female. Women who had reconstructive surgery indicated that the loss of feeling in the reconstructed breast was sexually troublesome and disappointing. Comparison of the qualitative data with quantitative data (triangulation) provided support for the finding of no significant differences in sexual functioning before women treated by lumpectomy versus mastectomy. CLINICAL IMPLICATIONS: Breast cancer affects many aspects of a woman's sexuality, including changes in physical functioning and in perception of femaleness. Informed healthcare decisions cannot be made until patients are knowledgeable about the sexual outcomes of cancer therapies. Healthcare professionals must acknowledge the scope of the impact of cancer treatment on sexuality, include such information as part of the informed consent process, and provide appropriate education and referral. PMID- 9397703 TI - Breast cancer detection by daughter of women with breast cancer. PMID- 9397704 TI - Transportation as a barrier to cancer treatment. AB - PURPOSE: Patients with cancer must overcome many psychological, social, and economic barriers to obtain needed treatment. Because of the need for repeated visits for cancer treatment on either an outpatient or an inpatient basis, one of the major issues that patients with cancer must confront is that of arranging for transportation to care. METHODS: This study compares the distance and mode of transportation to radiotherapy and chemotherapy and perceptions of transportation as a barrier to care among white, black, and Hispanic cancer patients receiving treatment from a consortium of cancer treatment facilities in Texas. A mail questionnaire was developed to assess the perceived barriers to cancer treatment for patients who had been diagnosed clinically with breast, colon, cervical, or prostate cancer, or lymphoma between 1989 and 1993. A total of 910 surveys were mailed to prospective participants. Of the surveys mailed, 593 were returned, yielding a 65.2% response rate. By race, the respondents included whites (42%), blacks (40%), Hispanics (15%), and Asian-Pacific Islanders (3%). Two respondents were 17 years of age; the remaining respondents were 18 years or older. RESULTS: This study shows that some patients may forgo needed treatment because of problems with transportation. This was perceived as an issue more for minority patients than for white patients. Black and Hispanic patients consistently reported that barriers such as distance, access to an automobile, and availability of someone to drive them to the treatment center were potential major problems. The distance to the facilities was farther for whites than for blacks and Hispanics. Patients generally had to travel farther for chemotherapy than for radiotherapy. CLINICAL IMPLICATIONS: Patients, particularly minorities, may opt to forgo needed care in the absence of available and affordable means of transportation to treatment facilities. These findings demonstrate the need for healthcare providers to be aware of the transportation problems that patients with cancer experience in obtaining treatment. Healthcare providers must work with patients, their families, and volunteer agencies in the community to facilitate transportation to cancer treatment services. PMID- 9397705 TI - What providers should know about community cancer control. AB - PURPOSE: The authors describe a framework for developing an effective, community focused cancer control program. OVERVIEW: Progress in the application of cancer control interventions has proven to be quite variable across different populations and communities. The Kentucky Cancer Program, developed under joint sponsorship of cancer centers at two state universities, has been using a model that appears to provide a high degree of sensitivity to community-specific problems and solutions. The Kentucky four-step model includes 1) using data from a population-based cancer registry and other sources to identify problems; 2) ensuring community involvement with providers in selecting the target population and developing the intervention strategy; 3) implementing the intervention plan; and 4) using cancer registry and other data to evaluate the impact of this intervention. CLINICAL IMPLICATIONS: This framework may be useful to providers as they try to balance the goals of their clinical practice and the goals of community cancer control. Developing an effective, community-focused cancer control program requires that providers gain a solid knowledge about their community. The depth and richness of that knowledge base is enhanced by the active participation of community members as they collaborate with the providers on planning and implementing cancer control activities. PMID- 9397707 TI - Cancer resources for providers in the rural community. AB - Rural residents with cancer face many challenges--lack of specialized local care, cost, and travel requirements--in receiving high-quality cancer care. Rural physicians, nurses, social workers, and pharmacists have limited resources to provide or coordinate the quality of care needed for these patients. Although resources are sparse, creative collaboration and capitalization on community cohesiveness may provide patients and healthcare professionals with needed resources in rural areas. Transportation for cancer patients, for example, might be obtained through local community organizations. Healthcare professionals may use various national cancer and rural health organizations and Internet sites as resources for updated oncology literature and for information on upcoming oncology conferences and partnerships with urban cancer care providers. PMID- 9397706 TI - An office systems approach to cancer prevention in primary care. AB - PURPOSE: The provision of preventive services holds a central place in primary care. Achievement of prevention standards offers a challenge. The authors address the efficacy of an office systems approach to improving cancer prevention and early detection services, provide a guide to assessing the appropriateness of office systems dissemination in practices targeted for improvement, and describe the range of dissemination strategies available. OVERVIEW: Preventive service office systems depend on establishing practice routines, using tools such as flow sheets, and sharing responsibilities among practice clinicians, staff, and patients. Systems have been shown to be efficacious in a variety of settings. Computers provide a significant enhancement to paper-based tools. Some practices develop office systems themselves, whereas others require external support. Before attempting to disseminate preventive services offices systems, disseminators should ensure that adequate assistance can be provided, that assistance follows a format that is acceptable to target practices, and that target practices are receptive to assistance and able to cooperate. Dissemination strategies include journal articles, continuing education programs, manuals and tool kits, facilitation, and academic detailing. The relative expense and efficacy of these approaches require further assessment. CLINICAL IMPLICATIONS: Office systems hold promise in enhancing provision of cancer prevention services in primary care. The practice should be approached as a team, and should include practice clinicians as well as nonclinical staff members. Current research should provide answers over the next few years about the cost-effectiveness of various approaches and the most feasible ways to promote dissemination to practices that need it. PMID- 9397708 TI - Collaborative practice with children and parents. Enhancing preparation for and management of cancer treatment. PMID- 9397709 TI - Anastrozole. An effective, second-line hormonal treatment for advanced breast cancer. AB - Because advanced breast cancer is not curable, the goal of treatment is to prolong survival yet maintain an acceptable quality of life. Anastrozole offers another option for second-line hormonal therapy in postmenopausal women with advanced breast cancer. This recently approved selective aromatase inhibitor is as efficacious as other available hormonal therapies for this indication and appears to offer an advantage with regard to adverse effects and ease of administration. PMID- 9397710 TI - Shareholders take stock in nursing's future. PMID- 9397711 TI - Balancing calcium and phosphorus levels in chronic renal failure patients. AB - Patients with chronic renal failure (CRF) can develop problems such as lethargy, tetany, and muscle spasms, which can increase their morbidity and mortality. Because of their non-functioning kidneys, patients with CRF require in-depth and comprehensive monitoring of calcium (Ca) and phosphorus (P) levels. This article presents advanced nursing actions and critical thinking strategies for use by the critical care nurse when caring for patients with CRF. PMID- 9397712 TI - Application of a transactional model of stress and coping with critically ill patients. AB - Critically ill patients are exposed to many physiologic and environmental stressors, which can result in deleterious physiological and psychological effects. Stress and coping within Lazarus and Folkman's transactional model is used as a basis for describing patient responses in critical care. The authors propose specific strategies to reduce stress and maximize coping in the critically ill patient. PMID- 9397713 TI - New neurointerventional therapies for stroke patients. AB - New neurointerventional procedures affect patient outcomes following stroke. Intraarterial thrombolysis, cerebral angioplasty, coils placed inside aneurysms, and vessel occlusion with embolic agents serve as options for stroke patients with conditions not as amenable to traditional surgical or medical management. By understanding these new neurointerventional therapies, critical care nurses can utilize a focused neurological assessment to intervene and maintain perfusion to the brain following the procedures. PMID- 9397714 TI - Recognizing the potential for violence in the ICU. AB - Violence in health care institutions, specifically critical care units, throughout the country is increasing at an alarming rate. This problem is not confined to the urban areas but also encompasses rural regions. This article assists the nurse in identifying the potential for violence by patients, family, or visitors while they are in the critical care unit. The author describes strategies that have been successful in controlling violence in the ICU and suggests steps nurse educators and managers can take to reduce the risk of violence in their units. The article is followed by Education STATPack material that can be used by current subscribers for an inservice. PMID- 9397716 TI - In the final analysis, people ... not$$$'s provide "care". PMID- 9397717 TI - Abbreviations that cause injury, complicate communication and may kill!. PMID- 9397715 TI - Developing a multidisciplinary brochure to teach patients and families about life sustaining treatments. AB - In an effort to comply with the Patient Self-Determination Act hospitals have scurried to develop patient education emphasizing patients' rights to accept or refuse medical treatments and to complete Advance Directives available in their state. Despite efforts at patient and family education, nurses report family difficulty in making end-of-life decisions because they have not fully clarified their values with regard to life-sustaining treatments, making decisions in crisis situations difficult. A brochure that discusses life-sustaining treatments along with Advance Directives is needed to help families understand this information before they reach a critical situation where anxiety makes decision making difficult. The critical care nurse, clinical nurse specialist, or manager can initiate a multidisciplinary task force to coordinate the development of a brochure used to increase community awareness. The purpose of this article is to help staff develop a brochure that can address these issues. PMID- 9397718 TI - Heads up nurses! "Prospective payment" is coming to your Medicare skilled nursing facility. PMID- 9397719 TI - Teaching the snail to fly: affecting change in long-term care nursing staff. PMID- 9397720 TI - Developing/implementing subacute programs in long-term care. AB - Subacute care is a new and rapidly growing area. The industry is still constantly changing. Prior experience in hospitals or long term care is not enough to ensure success in this new arena. Success depends on giving careful consideration to the corporate mission, staff selection and orientation, and to staff training. As the push continues toward providing comprehensive care programs delivered at a negotiated cost and within a set time frame, new roles for SAC facilities will evolve. In developing their roles, SAC facilities have much to gain from the prior experiences of their staff. However, staff members must also be willing to adapt, and to learn new ways of doing their jobs. Success goes to those who master the intricacies of this complex, challenging and dynamic industry. PMID- 9397721 TI - [Responsibility matters--registration is no threat]. PMID- 9397722 TI - [Midwives' matters in the Liability Committee]. PMID- 9397723 TI - [Responsibility matters--this is a scientific council]. PMID- 9397724 TI - [Responsibility matters--pride--accountability--report]. PMID- 9397725 TI - [Responsibility matters--a liability case in Denmark]. PMID- 9397726 TI - [Alternative Birth-Center Clinic in South Hospital]. PMID- 9397727 TI - Professionalism and ministry. Can we keep our balance. PMID- 9397728 TI - Nursing's moral standard. PMID- 9397729 TI - The price of autonomy. PMID- 9397730 TI - A call to servanthood. PMID- 9397731 TI - Spiritual debridement. PMID- 9397732 TI - Managing to care under managed care. PMID- 9397733 TI - Sadness, survival & hope. PMID- 9397734 TI - Not on my shift. PMID- 9397735 TI - The towel the bath. PMID- 9397736 TI - I'm afraid I'll cry. PMID- 9397737 TI - Fleshing out a nursing theory. A Christian conceptual model. PMID- 9397739 TI - Influencing our profession through continuing education. PMID- 9397738 TI - My search for meaning & purpose. PMID- 9397740 TI - Encouraging spiritual care. PMID- 9397741 TI - Spiritual care. Responding to an invitation. PMID- 9397742 TI - The gift of time. PMID- 9397743 TI - Mentored to mentor. PMID- 9397744 TI - Affiliated-individuation as a mediator of stress and burden in caregivers of adults with dementia. AB - The purpose of this study was to test a midrange model based on Modeling and Role Modeling Theory. The self-care resource of affiliated-individuation was tested to determine its mediational qualities between the stress and burden of caregiving and caregiver life satisfaction. The sample of 107 family caregivers of adults with dementia completed measures of stress, burden, affiliation, individuation, and life satisfaction. All variables were correlated in the expected direction. Affiliated-individuation decreased the effects of stress and burden on caregiver life satisfaction, lending support for the theory-based model. PMID- 9397745 TI - Potential benefits of pet ownership in health promotion. AB - Pet ownership provides an opportunity to improve health. A pet may become a stimulus for exercise, reduce anxiety, and provide an external focus of attention. Pets are also a source of physical contact and comfort and may decrease loneliness and depression while promoting an interesting lifestyle. The benefits of pet ownership are consistent with the health promotion and disease prevention goals outlined in Healthy People 2000. These goals include (a) increasing physical activity and fitness and (b) improving mental health and preventing mental disorders. Assessment of pet ownership and discussion of potential health benefits facilitates a holistic understanding of our patients and ourselves. PMID- 9397746 TI - Cultural meanings of childbirth: Muslim women living in Jordan. AB - This descriptive, ethnographic study focuses on the experience of childbirth for Muslim women living in Jordan. Thirty-two childbearing women were interviewed in the early postpartum weeks. The audiotaped interviews were transcribed and translated. Themes were identified from the rich, narrative data. Motivations for having children, as well as what constitutes the motherhood feeling, were described. Themes also included the importance of relying on God or Allah for support in childbearing and child rearing. A strong sense of the spiritual dimensions of giving birth within women's traditional, religious, and cultural context was identified. Findings from this study provide insight into the meanings of childbirth for Muslim women living in Jordan. These meanings assist nurses in providing culturally competent care. PMID- 9397747 TI - Weight and weight-related distress after childbirth: relationships to stress, social support, and depressive symptoms. AB - The aim of this study was to explore whether women's psychosocial context was related to weight status 1 year after childbirth. A survey sample of 149 women provided data on life-event stress, social support, and depressive symptoms; and three weight variables: body mass index, weight gain, and weight-related distress. Of the women, 32 (22%) reported gains of > or = 5 kg and 50 (34%) met the criterion for high depressive symptoms. Low social support, low income, and high depressive symptoms were related to higher weight gain. Women with gains > or = 5 kg reported high depressive symptoms (53% vs. 28%) more often than women with lesser gains. Women who reported lowered self-esteem because of weight also had higher depressive symptoms, body mass indexes, and weight gains than women with increased or unaffected self-esteem. This study points to the importance of incorporating women's psychosocial context into counseling about weight management after childbirth. PMID- 9397748 TI - Expanding self-awareness through exploration of meaningful experience. AB - In a phenomenological investigation of nurses' meaningful experiences with patients, the author continued investigation of caregiver/patient interaction reported in two previous studies of confirmation and exclusion. Exploration of meaningful experience is presented as an avenue for greater self-awareness for the caregiver. Findings include description of study participants' lived meaningful experiences and the essential structures of experiencing meaningfulness. The lived meaningful experiences were characterized by intense emotion, implicit experiencing, and relating. The essential structures of experiencing meaningfulness were found to be a temporal process of reflecting and articulating the discovery of self and the function of meaningful experience as a template for both future and past experiences. Expanded self-understanding through exploration of meaningful experience is discussed for its significance in nursing practice and education. PMID- 9397749 TI - The case of Baby M: nursing care in an ethical wilderness. AB - This is a reflective case study of an infant with Down Syndrome and a potentially fatal cardiac defect. It is a story of hope and loss, of silence and learning to speak, and of relinquishing space and standing ground. The purpose of this article is to explore the conflicting claims a neonatal intensive care (NICU) nurse faces in caring for critically ill infants. The questions of "Who speaks?" and "Who listens?" are addressed. The concepts of women's moral development and a nursing definition of voice are included. It is proposed that the conventional feminine voice and the embodied knowledge so integral to expert nursing actually draw strength away from the voice that needs to be permitted into the circle of decision makers when ethical issues are raised in the NICU. PMID- 9397750 TI - [It is great to be a midwife]. PMID- 9397751 TI - [What is the main principle when new employees are appointed--the title and formal qualifications or the skillful competence?]. PMID- 9397752 TI - [On titles and qualifications]. PMID- 9397753 TI - [Finnish maternity care and Chinese medicine]. PMID- 9397754 TI - [Zonal therapy courses are popular]. PMID- 9397755 TI - [Promises on the way to a better humanity]. PMID- 9397756 TI - [From the shadow to the light--the history of giving birth]. PMID- 9397757 TI - [Abortion--a subject kept in the dark]. PMID- 9397758 TI - [The rights of fetus and mother--conflict or harmony?]. PMID- 9397759 TI - [Finnish prenatal care and Chinese medicine. Acupuncture and other treatments]. PMID- 9397760 TI - [Ideal entrance of the child into the world]. PMID- 9397761 TI - [Breast feeding--secondary performance or a matter of course?]. PMID- 9397762 TI - [Confidence on every level]. PMID- 9397763 TI - [Depression after childbirth]. PMID- 9397764 TI - [Supportive measures for tired and depressed mothers in a Helsinki home for unmarried mothers]. PMID- 9397765 TI - [HIV-infections in Finland]. PMID- 9397766 TI - [Finnish prenatal care and Chinese medicine]. PMID- 9397767 TI - [Private midwife as the muse of the woman giving birth]. PMID- 9397768 TI - [The New Zealand Model--the 10-year path from a "doctor-nurse" to a self-employed professional]. PMID- 9397769 TI - [Breast program as the basic program for early recognition of breast cancer]. PMID- 9397771 TI - [Active birth and birth in water in Finland]. PMID- 9397770 TI - [Successful guidance?]. PMID- 9397772 TI - [Coli and children]. PMID- 9397773 TI - [Unsatisfactory sucking behavior at the breast. Detecting--preventing--treating]. PMID- 9397774 TI - [Money does not make us happy, gold does. An initiative by the coworkers in the neonatal nursery]. PMID- 9397775 TI - [Nasal continuous positive pressure respiration as a respiratory aid in small premature infants. From the physician's viewpoint]. PMID- 9397776 TI - [Nasal continuous positive pressure respiration as a respiratory aid in small premature infants. From the nurse's viewpoint]. PMID- 9397777 TI - [Studying to be a nurse--report on the nursing diploma program in Osnabruck]. PMID- 9397778 TI - [Experiences with nursing education in Frankfurt/Main]. PMID- 9397779 TI - [The importance of kinesthetic infant handling in pediatric nursing]. PMID- 9397780 TI - [160 years of pediatrics in Austria]. PMID- 9397781 TI - [Development of pediatric nursing in Austria. An historical retrospective]. PMID- 9397782 TI - [The parents' work at the Glanzing Pediatric Department]. PMID- 9397783 TI - [Sudden infant death]. PMID- 9397784 TI - [Terminal care is obligatory for physicians and nurses]. PMID- 9397785 TI - [Special features in the postnatal adaptation of newborns after cesarean section]. PMID- 9397786 TI - [Special features in the postnatal adaptation of newborns after cesarean section- nursing aspects]. PMID- 9397787 TI - [Psychosocial care of the parents in neonatology. Report of experiences]. PMID- 9397788 TI - [In the shadow of a sick sibling]. PMID- 9397789 TI - [Caring for a newborn with meningomyelocele]. PMID- 9397790 TI - [Treatment of persistent pulmonary hypertension of the newborn by nitrogen monoxide inhalation]. PMID- 9397791 TI - [Outpatient oncological nursing service. Description of a project]. PMID- 9397792 TI - [Interview by the Nursing Society with Elisabeth Oltrogge. Profiles in pediatric nursing--personalities and challenges]. PMID- 9397793 TI - [10 years Charlottenburg Birthing Center. Report on 10 years of births in the 1. European birthing center in Berlin, Klausenerplatz]. PMID- 9397794 TI - [Accident insurance legislation. Regulations for accident prevention contain important rules of behavior]. PMID- 9397796 TI - [Kneipp year 1997]. PMID- 9397795 TI - [Management in the nursing service--workshop for nursing personnel in the European Surgical Institute]. PMID- 9397797 TI - Sexuality and the spinal cord injured. PMID- 9397798 TI - Sexual health: every nurse's concern. PMID- 9397799 TI - Sexual healing. PMID- 9397800 TI - Straight talk and humour adolescent sexual issues. PMID- 9397801 TI - Learning about sexual health. PMID- 9397802 TI - Respoking the wheels of health. PMID- 9397803 TI - Sex in residential care facilities. PMID- 9397804 TI - Save your back! PMID- 9397805 TI - Breast cancer is the most common cause of cancer death in women. PMID- 9397806 TI - Legal issues for nurses. PMID- 9397807 TI - A project to obtain clearer work-related injury and illness data for the health industry. PMID- 9397808 TI - Lighting the Way. The Enrolled Nurse Professional Association of NSW Inc. PMID- 9397809 TI - Sexuality is a highly complex phenomenon. PMID- 9397810 TI - Nurse in profile. Robyn Hately. Interview by Kimberly O'Sullivan. PMID- 9397811 TI - Ethical issues in nursing education. PMID- 9397812 TI - Writing and evaluating computer-based training programs. AB - Publishers are preparing for the hitech future and authors and editors need to also. Publishers know that book sales may be down in the future, while computer program sales are expected to increase. Most book publishers have added media divisions to position themselves for the future. Authors and editors can also shift from writing or editing books to developing computer-based training programs. This author tells you how to write, edit, or evaluate the new computer based training programs. PMID- 9397813 TI - Book reviewing: keeping the audience in mind. AB - Writing book reviews is an excellent way to start your publication experience and develop a relationship with a journal editor. Most nursing journals include book reviews, and some even select editorial board members from book reviewers who have demonstrated their ability to write concise, honest reviews and submit them on time. This author includes tips on writing a high-quality review by keeping the audience in mind. PMID- 9397814 TI - Fax etiquette for nurse authors and editors. AB - Is the facsimile (fax) machine really as great as it seems? Yes, but there is a potential for its misuse. Like all equipment, the fax machine is a tool that needs to be used wisely. This article describes the Do's and Don'ts of using the fax machine to communicate between authors and editors. Tips in this article will help authors and editors to correspond smoothly by fax and use new fax equipment options. PMID- 9397815 TI - Is your co-author relationship on the rocks? How to know and what to do. AB - Co-authorship can be successful, but there are pitfalls to avoid so that a positive relationship does not become a disastrous one. Even experienced co authors have to review and work on their relationship to make sure they complete the project and retain a collegial relationship. In a past article this author described how to develop the co-authorship relationship (Heinrich). In this article she describes how to identify and deal with co-authorship problems when they occur. PMID- 9397816 TI - Checking references: tips for reviewers. AB - When I was a new reviewer, I was uncertain if I should check some, all, or none of the references in the manuscript. I was equally uncertain, however, if anyone was ultimately accountable for this responsibility. Reviewers must assume responsibility for checking references because "the most critical part of the review process is to check the accuracy of the content." (Brooks-Brunn, 1993, p. 4). The reviewer's three main responsibilities include checking for (1) a reference for every citation (2) accuracy of reference content (3) accuracy of cited material (Kirchhoff, 1995). PMID- 9397817 TI - Successful publishing in nursing: debunking some common myths. AB - Myths can serve useful functions for a group or society. The myths related to success in publishing, however, do not seem to serve a useful function, and may operate as barriers or excuses that unnecessarily hinder success in publishing. In this article, some of the common myths associated with publishing in nursing are examined for their validity and for evidence of support for the myth in the literature. PMID- 9397818 TI - Submitting your manuscript on disk: a steep learning curve for authors and editors. AB - Many nursing journals are switching to state-of-the-art manuscript production systems. Most of these new systems include computerized typesetting, so some journals are requesting disk submission of manuscripts. However, computer disk submission is not without difficulties. Editors and authors alike are experiencing a fairly steep learning curve before developing a system where the author's disk can work in the publisher's production system. This article describes how authors and editors can collaborate on the disk submission system for nursing publications. PMID- 9397819 TI - Nurse writers internet and World Wide Web sites. AB - Nurse writers can use the Internet to find vast amounts of current information on topics for professional articles. The following excerpts are based on concepts in the Second Edition of the authors' book, How to Write and Publish Articles in Nursing, published by Springer Publishing Company. The Second Edition will is now available. PMID- 9397820 TI - Discovering medical products on the Web. PMID- 9397821 TI - Myths & facts ... about aging. PMID- 9397822 TI - Performing Allen's test. PMID- 9397823 TI - Taking a risk for Ms. Stinson. PMID- 9397824 TI - Managing pain during chest tube insertion. PMID- 9397825 TI - Removing catheters. Managing freezes and fractures. PMID- 9397826 TI - Actionstat. Acute pulmonary edema. PMID- 9397827 TI - Looking out for adverse drug reactions. PMID- 9397828 TI - Taking diabetes new look at an old adversary. PMID- 9397829 TI - Ventricular tachycardia. PMID- 9397830 TI - The art of nursing. PMID- 9397831 TI - Pediatric patient care. PMID- 9397832 TI - Caring for patients with benign prostatic hyperplasia. PMID- 9397833 TI - Nurse's guide to common postoperative complications. PMID- 9397834 TI - Closed suction wound drainage. PMID- 9397835 TI - Looking the other way. PMID- 9397836 TI - Breakfast in the old country. PMID- 9397837 TI - The nurse executive: leading beyond traditional boundaries. PMID- 9397838 TI - Profiles of leadership: a dialogue with two nursing revolutionaries. AB - Rebels break the rules and revolutionaries rewrite them because the old rules are no longer adequate. These revolutionary leaders examine their life journeys and highlight issues and events that prepared them for nontraditional nursing roles; that of consultant and CEO. Patterns of thought and behavior nested within their stories provide a rich map for the contemporary nurse seeking to expand their view of the health care landscape. PMID- 9397839 TI - My labyrinth. AB - Each of us is taking a professional journey. We use our scholarship and our relationship building skills to learn and mentor, experience and contribute, and lead and follow. Our creativity, flexibility, and adaptability provide us with a "centering" that enhances our success, but our only reason to pursue the journey with vigor is to create a better environment for professionals to practice and for patients to receive care. Let us know ourselves so we can ground the journey with integrity. PMID- 9397840 TI - Nursing journalism leadership. AB - How is leadership exemplified in the role of nurse editor? In this article, the editor of two nursing journals describes the evolution of her editorial role with a focus on job analysis and leadership requirements. Highlighted are catalyzing events that shaped the author's editorial vision and leadership style. Factors for success in the role are presented to help those who want to assume editorships. PMID- 9397841 TI - The private practice of nursing: the gift of entrepreneurialism. AB - The demands of the entrepreneur extend the opportunities and creativity of the nurse. The characteristics of independent practice are now the expectations of the role of every nurse. Understanding the gifts of the entrepreneurial experience helps facilitate the growth of nurses and their practice. PMID- 9397842 TI - Leading beyond traditional boundaries: a community nursing perspective. AB - As a nursing leader, I frame my personal and professional life from the perspective of community. I place high value on health, relationship and the art of nursing. As Director of Community Outreach for an integrated delivery network, I use these values to strengthen nursing's contribution to an integrated delivery network and managed care contracts. Most recently, my personal and professional life has been shaped by traditional Indian medicine and my 10-year experience in Carondelet's community-focused nursing practices. PMID- 9397843 TI - Nurse leaders: roles driving organizational transition. AB - Transition leaders have developed in our organizations. These roles allow nurse leaders to combine the knowledge of the customer, the art of nursing, and the chaos of health care in order to pioneer new strategies. These roles are flexible, either focused or broad, and change in scope and direction. The teams we lead are diverse in team experience, seen as disconnected from operations, and struggle with exceedingly defined or hardly evident boundaries. Methodologies add structure and foster active participation; they are fast, decisive, compressed, creating results. "Change-magic" assists in making decisions and influencing the organization to "get the job done." PMID- 9397844 TI - Determining if shared leadership is being practiced: evaluation methodology. AB - This article explores the use of a cognitive information-processing structure, mental models, to assess an organizational management change to shared leadership (SL). The purpose of this study was to describe the mental models of critical care nurses after the implementation of an SL management model. Scenarios based on daily clinical situations and measuring three SL concepts (empowerment, accountability, and partnership in decision making) were posed to critical care nurses in personal interviews. By using this evaluation methodology, nurse administrators can determine to what extent a new management change has been incorporated into daily nursing practice and in what areas to target further education and resources. PMID- 9397845 TI - Executive journey: from 1,300 FTEs to none. AB - The story of how one woman, Mary Jane Mastorovich, traveled the journey from the traditional role of a nurse executive, responsible for 1,300 full-time equivalents (FTEs) and an annual budget of $166 million, to that of a major change agent in a system-wide redesign effort. Her travels and the lessons she learned along the way give insight into how to develop strategies and methods for personal and professional transformation. Her story also illustrates the balance between positional power and the power of influence as we face the roller coaster ride that will be health care delivery in the next century. PMID- 9397846 TI - Nurse executives in the 1990s: empowered or oppressed? AB - Nursing executives are at the center of change. They are involved in the reenginnering of the workplace and often implement new procedures, in addition to downsizing staff. This article explores oppressed group behavior and its implications for nursing executives at this time of change in the health care system. The purpose of this exploration is to encourage a dialogue about its relevance in order to enhance empowerment and to break the cycle of oppression. PMID- 9397847 TI - Is nursing under new management or is it a matter of semantics? AB - Managed care, in addition to being driven by the hegemony of market concerns, has truly bitten into the clinical hegemony. Nursing has recognized this new order and is attempting to manage the situation with new paradigms before other non nursing entities do. The issue for nursing has become how to allow the patient to drive the health care delivery system while preparing for change and not fearing nursing's loss of preeminence as a clinical authority in its own right. At the same time, nursing must protect a valued and needed profession. PMID- 9397848 TI - The nurse executive in the 21st century: how do we prepare? AB - A revolution is occurring in health care. Hospital restructuring, mergers, and closures are occurring at an unprecedented rate. The role of the nurse executive is being scrutinized as never before. There is considerable debate on what is the best educational preparation for nurse executives given their increasing management and fiscal responsibilities in organizations today. This article reviews the evolution of graduate education in nursing administration as well as current research on graduate education for nurse executives. Job trends for nurse executives in industry and health care are also explored. PMID- 9397849 TI - New RNABC president seen as a mentor for nurses. PMID- 9397851 TI - New ways of doing things. PMID- 9397850 TI - The VON at 100. A century of caring. PMID- 9397852 TI - Sharing the challenge as volunteers. PMID- 9397854 TI - Care for your community. PMID- 9397853 TI - Partners in health. PMID- 9397855 TI - New Code provides guidance for ethical decision-making. PMID- 9397856 TI - Nurse clinician as learning resource. Interview by Helen Griffiths. PMID- 9397857 TI - Continual learning. Interview by Helen Griffiths. PMID- 9397859 TI - The disaster at Green Park. PMID- 9397858 TI - How far is too far. PMID- 9397860 TI - Home care of persons with AIDS. PMID- 9397861 TI - The role of nurses in crisis intervention. PMID- 9397862 TI - Nursing management in critical care. PMID- 9397863 TI - New trends and old traditions in educational technology: video and beyond. PMID- 9397864 TI - Broadening the staff development department. PMID- 9397865 TI - The evolving age-related competency: a potential type-one trap. PMID- 9397866 TI - Beating the mandatory blues: using the FOCUS PDCA process. AB - By October 1995, a total of 863 nursing employees fulfilled their annual requirements, for a compliance rate of 94.4%. Additionally, 178 employees from various ancillary departments such as radiology, dietary and food service, rehabilitation, and social service also attended. Therefore, we were able to use our program beyond the nursing department and provide a hospital-wide educational service. Overall, the poster presentations were highly successful and the benefits to the institution included the following: 1. An increase in employee satisfaction because the average time spent away from the patient care unit was substantially decreased. 2. An increase in instructor availability to meet the educational needs of the nursing staff on individually assigned units. 3. An increase in instructor accessibility to identify and meet the learning needs of new staff members. PMID- 9397867 TI - Student support. PMID- 9397868 TI - Competition in health care calls for creativity. PMID- 9397869 TI - Innovative collaboration to prevent repeated adolescent pregnancies. AB - Nurse educators from a university setting and staff from the county health department collaborated to establish an innovative program to prevent repeated pregnancy in adolescents. Called Dollar-A-Day and patterned after the original in Denver, CO, the program was operated jointly for 5 years and today continues to operate under the auspices of the health department. Success of the venture is attributed to use of skills in assessment, building, managing, and evaluating, as described by Loxley (1997). These elements were used to construct a context for collaboration. PMID- 9397870 TI - Why must a clinical specialist be a nurse practitioner? PMID- 9397871 TI - Concerns of graduating baccalaureate nursing students. AB - Senior baccalaureate nursing students (N = 397) just weeks away from graduation were asked to complete the "Worry Clinic Form." This form invites subjective responses from the students' perspectives about their "current" and "greatest-to date" professional concerns. Content analysis was used to determine the categories of these concerns. Categories of both current and greatest-to-date professional concerns were (1) functioning as a competent registered professional nurse and (2) the environment of work. PMID- 9397872 TI - Academic misconduct in schools of nursing. PMID- 9397873 TI - Cooperative learning in the clinical setting. AB - The modern clinical practice setting presents nurses with challenges about which they must think critically and develop increasingly autonomous problem-solving approaches. It is essential to provide nursing students with opportunities to practice critical thinking so that they can develop this crucial skill. Cooperative learning strategies are interactive teaching methods that stimulate students to think critically, communicate effectively with peers, and accept responsibility for learning through group process activities. Group care planning is one such cooperative strategy that also promotes a positive attitude about care planning and sharpens time management skills. Cooperative assessment and care planning foster the development of critical thinking and effective problem resolution, preparing students for patient care problems they will likely encounter in future positions. PMID- 9397874 TI - Outcomes of occupational stressors on nurses: chronic fatigue syndrome--related symptoms. AB - Considering the types and number of occupational stressors involved in caring for patients, nurses may represent a population at high risk for physical illnesses. A sample of 3400 nurses who belong to a statewide or a national nurses organization were randomly chosen for participation. Of this group, 202 reported 6 months or more of debilitating fatigue and completed a three-page questionnaire assessing symptoms related to chronic fatigue syndrome (CFS) and comorbid medical conditions. This group (N = 202) was mailed a follow-up questionnaire 1 year later that reassessed symptoms of CFS and occupational stressors. Many sampled nurses reported a high degree of occupationally related stress but did not report CFS symptoms; however, perceived exposure to the threat of an accident as a nurse and poor physical working conditions were significantly related to symptoms reported. These findings are consistent with previous research. PMID- 9397875 TI - Clinical process learning to improve critical thinking. AB - A six-step process-learning strategy model serves as a framework for nursing students to critically analyze situations encountered during their clinical practice experience. Stephen Brookfield's four components of critical thinking and culturalization themes relate well to how nurses learn and experience critical thinking and serves as the model's organizing framework. This learning strategy has implications for all nurse educators because it incorporates the realities of nursing practice, merges nursing education with practice, involves students in affective, cognitive, and psychomotor domains of learning, and prepares graduates to function in dynamic and complex health care systems. PMID- 9397876 TI - Symbols of our trade: examining our relationships with work. PMID- 9397877 TI - Cleveland cancer advocates develop regional resource directory. PMID- 9397878 TI - Risk assessment and risk-based programs of prevention in various settings. AB - An assortment of screening tools exist to estimate risk for pressure ulcer development. The Braden Scale has undergone testing in several settings which guide users in answering the following questions: Who should assess risk? Does cut-off score differ by setting? Does timing of assessment vary by setting? Is formal risk assessment necessary? How does risk assessment fit into a program of prevention? Based on these studies, RNs are able to use the Braden Scale more reliably than Nurse Aides and LPNs. While the Braden Scale does not replace clinical judgement, its use will help caregivers of all levels to identify at risk patients and intervene for specific risk factors. Generally, all patients should be assessed upon admission and 24 to 48 hours later, followed by ongoing assessment. A formal risk-based program is effective in both reducing the incidence of pressure ulcers and the costs associated with prevention. Protocols can be developed to address each level of risk, with each level requiring more aggressive preventive modalities. Some investigators have also tied interventions to specific subscale scores. It is important that risk assessment and risk-based protocols such as the Braden Scale become a standard of practice in all healthcare settings. PMID- 9397879 TI - Bandaging in the treatment of venous ulcers: a European view. AB - The principal treatment of venous ulcers in ambulatory patients in bandaging. The physiological rationale for this treatment is to improve the venous hemodynamic abnormality caused by prolonged venous hypertension due to limb venous valvular incompetency. Correct bandaging results in the reduction of limb edema and ulcer healing. A number of different bandages and combined bandaging regimens are used in the treatment of venous ulceration. Recently the European Tissue Repair Society has defined different types of bandages based on their function into four groups: (1) Extensible bandage; (2) Elastic bandage; (3) Compression bandage; and (4) Support bandage. The ability of a bandage to produce a gradient compression starting at 35 to 45 mmHG at the ankle and reducing as the bandage approaches the knee is stated in the literature as a goal of bandaging. Our studies and others have shown that routine measurements of limb circumference as an indicator of edema reduction is another way of monitoring the efficacy of the results of bandaging since edema reduction and control is associated with improved ulcer healing. PMID- 9397880 TI - The non-healing leg ulcer: peripheral vascular disease, chronic venous insufficiency, and ischemic vasculitis. AB - The non-healing leg ulcer is examined by discussing three disease processes: peripheral vascular occlusive disease (PVOD), chronic venous insufficiency (CVI), and vasculitis. For PVOD, management decisions are based on risk factors and disease history. Comprehensive management includes the discontinuation of smoking, exercise conditioning and regulation of diabetes, hyperlipidemia, hypertension, and the appropriate application of anticoagulant/antiplatelet drugs. Methods of surgical management include bypass with autogenous or synthetic material in addition to reconstructive surgery with patch angioplasty or extra anatomic bypass, amputation, percutaneous transluminal angioplasty/stents, thrombolytic infusion, atherectomy, intraluminal ultrasound, and angioscopy. The optimal healing environment for all ulcers prevents contamination, pain, and fluid loss. In CVI, higher venous pressure in the veins of the lower limb during exercise results in ambulatory venous hypertension and ulceration. Various theories are associated with the disease and ulceration process; the classic treatment of elevation, ambulation, and compression for venous disease remains unchallenged. Diagnosis is based on history, physical examination, invasive venography, and/or non-invasive studies. Two groups of vasculitic disorders that share varying degrees of vascular inflammation and necrosis are arteritis (lupus, erythematosus, periarteritis nodosa, dermatomyositis) and blood dyscrasias (sickle cell disease, thalassemia). Leg ulcers associated with vasculitis are due to inadequate tissue oxygenation at the local level, are typically chronic, slow to heal, and commonly recur. PMID- 9397881 TI - Clinical evaluation: outcomes, benchmarking, introspection, and quality improvement. AB - Transforming the current event-driven reimbursement system into a quality powered marketplace will require clinical evaluation of how care is delivered. The managed care marketplace is evolving in three stages, from an event-driven, cost avoidance model, to which the concepts of "value" and "quality" are added, with the final addition of a more "public health" focus. Clinical evaluation is a scientific process of outcomes assessment, clinical guidelines, and benchmarking. This process was applied to a hospital-based outpatient wound clinic, leading to a determination that the overall clinic Kaplan-Meier median time to healing could be improved. Two groups of patients were studied, 141 retrospectively from 1993 to 1994 and 57 prospectively in 1995. While there was no significant difference in the percentage healed between the groups, a significant difference in the median times to healing was revealed, which was linked to antibiotic use. Even when antibiotics were used prophylactically, the median times for healing were elevated from those without infections. Introspection led to fewer patients receiving preventive antibiotics. The overall lower median time to healing curve in 1995 can be explained by this change in clinical practice. This quality improvement demonstrates the utility of the clinical evaluation process as the healthcare marketplace evolves. PMID- 9397882 TI - Timing issues for wound care research. AB - While certain issues related to time, such as time to healing, are frequently discussed in the wound care literature, other important time-related issues for conducting and interpreting wound research are only rarely addressed. Time presents serious philosophical and methodological challenges that can have a significant impact on the validity, reliability or rigor of a study and are, therefore, worthy of consideration. This article discusses four commonly encountered time-related problems for wound research (time-congruent research questions and designs, time specific measurement tools and protocols, retrospective versus prospective studies, and time sampling/sequential triangulation), presents examples to illustrate these problems, and posits some solutions. Five questions are offered as a guide for determining whether wound research tools and protocols adequately address time-related research issues. PMID- 9397883 TI - The Wound Intelligence System: early issues and findings from multi-site tests. AB - The purpose of this manuscript is to address a gap in our efforts to incrementally improve wound care practice through evidence-based practice. The Pressure Sore Status Tool (PSST) provides data to extend evidence-based practice beyond clinical trials and into the clinical area itself. The computerized PSST was evaluated over one year through over 70 beta sites. Two studies which were part of that evaluation period are described which give a comparative analysis of wound stage and PSST scores, and similarities and differences in wound characteristics of four types of wounds: arterial/ischemic ulcers, neuropathic ulcers, pressure ulcers, and venous ulcers. In the first study, a relationship between PSST scores and staging scores for the presenting wound was present, indicating promise for the utilization of the PSST as an alternative to staging scores for describing changes in wound status. However, in the second study, clear difference was not noticeable between the four wound types, suggesting that discriminations regarding wound type may not be able to be made from PSST assessments. The goal of these studies was to provide feedback on the use of the computerized PSST, thereby providing feedback based on objective outcomes of the practice of clinicians themselves. PMID- 9397884 TI - The road to developing standards for the diagnosis and treatment of venous leg ulcers. AB - In early 1996 the Venous Leg Ulcer Guideline was developed for the diagnosis and treatment of venous leg ulcers. In order to discuss the development of standards in general, and the Venous Leg Ulcer Guideline in particular, we first need to understand the difference between the following terms: Critical pathway, consensus statement, guideline, and standard. There are advantages and disadvantages to the use of guidelines. In the development of a guideline, endorsement by a respected colleague is important. Development of the Venous Leg Ulcer Guideline began with a consensus statement and then underwent review by a national advisory panel and national peer review through publication. A revised guideline has now been developed which will be tested in a pilot study for clinical efficacy, effect on cost, and impact on quality of life. Validation will require implementation in a prospective clinical trial. Diagnostic and Treatment Algorithm forms for the diagnosis and treatment of venous leg ulcers were developed as part of the preliminary testing of the guideline. Although guidelines do not substitute for good clinical judgement, they can encourage clinical judgement and help reduce fragmented care and the costs associated with inappropriate treatments. PMID- 9397885 TI - Issues in chronic wound care: where do we go from here? AB - Understanding of the healing process and factors that may affect it, has increased dramatically during recent years. Clinical application of this knowledge has improved care for many patients, and helped focus attention on information deficits. To better meet patient needs and increase awareness of the value of prevention, wound care clinicians and researchers are encouraged to include Quality of Life outcomes in their goals of chronic wound care. In the absence of the results of reliability, validity and feasibility studies of existing chronic wound care guidelines, clinicians need to be aware of their potential usefulness and limitations. When reviewing wound care costs or conducting studies, it is important to define cost-effectiveness as a function of outcomes and all relevant costs. Because chronic wounds occur in every care environment, these issues require the attention of all healthcare professionals, educators, payors and patients. Wound care professionals are encouraged to disseminate existing knowledge to all providers and recipients of care. PMID- 9397887 TI - Charge nurses entitled to vote in Union elections. PMID- 9397886 TI - Nurses have the courage to care. PMID- 9397888 TI - Nurse's duty to guard against sexual predators on staff. PMID- 9397889 TI - LA: improper administration of injection: nurse's failure to record "site & mode". PMID- 9397890 TI - PA: drug conviction triggers suspension: nurse charge board violated ADA. PMID- 9397892 TI - A familiar tune. PMID- 9397891 TI - Does workers' compensation cover on the job heart attack? PMID- 9397893 TI - Ethics in action. Your cancer patient's pain. PMID- 9397894 TI - The 1997 earnings survey. More benefits, with a price. PMID- 9397895 TI - Analyzing the Chem 7. PMID- 9397896 TI - Healing venous ulcers. PMID- 9397897 TI - PA catheter numbers made easy. PMID- 9397898 TI - Vitamins and minerals. PMID- 9397899 TI - Subacute care: yesterday's med/surg. PMID- 9397900 TI - When you go toe-to-toe over doctor's orders. PMID- 9397901 TI - Breaking free from sedative addiction. PMID- 9397902 TI - Who was I to judge? PMID- 9397903 TI - [Challenge for professional group? Treatment of chronically ill especially important]. PMID- 9397904 TI - [Are we really getting those new colleagues? Annual survey health care 1998]. PMID- 9397905 TI - [Needs of the people are determinant for nursing care--learning to think globally. Interview by Willem Zandbergen and Tonny van de Pasch]. PMID- 9397906 TI - [Hospital and university work together on quality--autonomous nursing in the NDU (Nursing Development Unit). Interview by Tonny van de Pasch]. PMID- 9397907 TI - [A week of chronic diseases. 2nd European Nursing Congress]. PMID- 9397908 TI - [Karin Spaink lives with MS for 10 years already--'When I am stable' I don't worry about anything. Interview by Toine de Graaf]. PMID- 9397909 TI - [To be chronically ill, life style and nurse's contribution]. PMID- 9397910 TI - [Nursing care of chronically ill still not optimal--rising above the disease image. Interview by Tonny van de Pasch]. PMID- 9397911 TI - [A special responsibility for nurses--the chronically ill in the hospital]. PMID- 9397912 TI - [2nd European Nursing Congress 5 through 8 October]. PMID- 9397913 TI - [Depression in the center--National Day for Public Mental Health]. PMID- 9397914 TI - [Nurses have trouble with inhalation therapy. 'Nurse, can you show us that puff?']. PMID- 9397915 TI - [Nurses can point out to parents the adverse effects for young children. Campaign against passive smoking]. PMID- 9397916 TI - [Pilot project limited to The Netherlands. Debate on Internet about nursing diagnosis]. PMID- 9397917 TI - [Not extending life is different from ending life. When the remedy against death only extends the dying process]. PMID- 9397918 TI - [Searching for sense and meaning together with the patient. Giving meaning and professional nursing practice]. PMID- 9397919 TI - [It remains different for the patient than for the nurse--the world behind the bed bath]. PMID- 9397920 TI - [Treatment compliance is the key word in combination therapy. Current image of AIDS (2)]. PMID- 9397921 TI - [Many requests for aid in the nurse's field. Nursing office hours for MS patients]. PMID- 9397922 TI - [Diagnosis in nursing. Prognosis: from expectation to goal-setting]. PMID- 9397923 TI - [Nursing care and total health care costs]. PMID- 9397924 TI - [Pressure sores in ICU patients--a literature review]. AB - The purpose of this literature study is to gain an insight into the information available on epidemiological aspects, specific factors of risk, risk assessment instruments and preventive measures with respect to pressure sores with adult ICU patients. There are indications that the incidence of pressure sores in the ICU is higher than the average incidence of 3-10% in hospital populations. This higher incidence may be accounted for by specific risk factors, such as history before admission to the ICU, severity of the disease, the risk of prevention measures, the use of special medicine and the nutritional state. Also structural factors, such as the accessibility of prevention means and the unambiguous usage of the registration of pressure sores, play a part. Risk assessment instruments are used to quantify the risk of pressure sores in order to support or evaluate the decision concerning preventive measures. For the ICU risk assessment instruments have to be developed, in which the specific factors of risk are processed. Related to preventive measures the effect of turning and re positioning the patient on the vital functions should further be investigated. If turning and re-positioning could be started as early as possible and in a responsible way, the demand for expensive measures, like special beds en special mattresses, will possibly decrease. The positive effect of these special beds on the prevention of pressure sores seams to be irrefutable. PMID- 9397925 TI - [The use of isolation in psychiatry--a literature study]. AB - This article reports on a literature review into the publications on the practise of seclusion. Attention will be paid to the moral debate on seclusion. The major part of the publications consider seclusion as a necessary intervention to manage problem behaviour. The first part of the article will consider definitional aspects and will result in concept clarification. The review shows differences at definitional aspects, motives for seclusion and patient characteristics. Data on frequencies, incidence and duration appear to be different. The experience of patients who are secluded are mostly negative, but positive reactions are also reported. Publications on the influence of the hospital characteristics to the use of seclusion seem to increase during time. Finally it is concluded that seclusion is an effective way to manage (potential) dangerous behaviour and that seclusion is an intervention which may create therapeutic conditions. PMID- 9397926 TI - [Ethics of nursing practice--ethics of care and ethics of rule?]. AB - Nurses' daily care for patients is imbued with moral questions. Ethics distinguish various movements that intend to solve the problem of how to deal with such questions. This article discusses two variants, namely ethics of rules and ethics of care. Ethics of rules present a rational model of thinking in which universal principles are applied to practical moral problems. According to ethics of care, an attitude of responsibility and involvement and, accordingly, the attention to the complex situation of a patient is considered moral. Both kinds of ethics are attached to a case from the practice of nursing care for oncology patients. The thinking of the nurses concerned is in line with ethics of care. It also involves principles of ethics of rules. These principles, however, only make sense in the complex situation of the patient. By virtue of this quality they are taken into consideration by nurses as one of the details from the context. This makes ethics of care function as a kind of 'breeding ground' for ethics of rules. PMID- 9397927 TI - [Breathing-related nursing care problems--a descriptive study]. AB - Research into nursing problems related to breathing that can occur by pulmonary patients and their relationship with literature on Nursing Diagnoses (further referred to as diagnoses) on this subject has been done on two nursing wards. Interviews have been held with eleven nurses working in direct patient care. From the acquired data twelve problems with their signs and symptoms as well as the possible etiology were found, each showing one or more similarities with diagnoses from the North American Nursing Diagnosis Association (NANDA). Similarities and differences between the NANDA diagnoses and the acquired data have been analysed. PMID- 9397928 TI - [Does the tension-filled relationship between theory and practice concern an interactional and/or a translational process?]. PMID- 9397929 TI - Thyrotropin-releasing hormone (TRH)-related peptides. PMID- 9397930 TI - PACAP, PACAP receptors, and intracellular signalling. PMID- 9397931 TI - Isolation and characterization of the Xenopus laevis cDNA and genomic homologs of neuropeptide Y. AB - We have isolated Xenopus laevis cDNA and genomic clones encoding the neuropeptide Y (NPY) mRNA and gene using a probe from the human NPY gene. The longest open reading frame in the cDNA encodes a peptide 76% identical to human prepro-NPY and 73% identical to rat prepro-NPY. The putative mature Xenopus NPY (XNPY) peptide is 94% identical to both human and rat peptides. A genomic clone containing 422 base pairs of 5'-flanking sequences and the 5'-end of the mRNA was also isolated. Primer extension analysis was used to map the transcription initiation site of the Xenopus NPY gene. Comparison of the 5'-flanking sequences of the Xenopus laevis, human, and rat NPY genes resulted in areas of high conservation, including the TATA box and the CT box previously shown to interact with Sp1-like proteins. Distribution of the Xenopus NPY message was analyzed by Northern analysis and RNAse protection. XNPY transcripts were not detected in whole developing embryo RNA, but were detected in adult frog brain RNA. We have also conducted preliminary studies of the XNPY promoter, utilizing an XNPY/chloramphenicol acetyl-transferase fusion construct. This study has demonstrated that Xenopus NPY shares a high degree of identity to its human and rat counterparts and that this homology extends to the gene, which contains similar cis-elements positioned near the transcription start site. PMID- 9397932 TI - Immunochemical mapping of human lutropin: I. Delineation of a conformational antigenic determinant. AB - Lutropin (LH), follitropin (FSH), thyrotropin (TSH) and choriogonadotropin (CG) are assembled of two non-covalently alpha (alpha) and beta (beta) subunits. We studied the discontinuous antigenic regions recognized by a monoclonal anti-hLH antibody designated as LH05 which binds to hLH, hCG and hTSH and does not cross react with either the free subunits or hFSH. LH05 and an antibody designated HT13, recognizing an epitope partly comprizing the alpha64-76 region, did not bind simultaneously to hCG. Furthermore, LH05 was unable to combine with an anti peptide antibody (LHP03) directed to residues 43-52 of hLHbeta. Thus, LH05 recognizes an epitope partly overlapping with those recognized by HT13 and LHP03. Using various hybrid molecules, we showed that the human alpha-subunit plays a critical role in the assembly of the epitope that, in contrast, contains amino acid residues conserved in the various beta-subunit of several species. Together, our results suggest that the amino acids Leu49-Pro50, Tyr59-Arg60 and Leu86-Ser87 in the hLHbeta and the alpha64-76 region are probably included in the epitope recognized by LH05 which appeared to be not accessible on the CG/LH receptor. PMID- 9397933 TI - Immunochemical mapping of human lutropin: II. Characterization of two monoclonal antipeptide antibodies reacting with the native beta-subunit. AB - To investigate the epitopes present on the beta-subunit of the human lutropin (hLHbeta) and their topographical relationship at the surface of the molecule, we produced two monoclonal antipeptide antibodies, designated LHP03 and LHP04, capable of binding to the radiolabeled 125I-hLHbeta and directed to the 43-52 and 110-117 regions of the hLHbeta, respectively. Analysis of the accessibility of the epitopes on hLH and on the beta-subunit of human chorionic gonadotropin (hCGbeta), equine LH (eLHbeta) and ovine LH (oLHbeta) indicated that: (i) LHP03 binds to both the free hLHbeta subunit and dimeric hLH whereas LHP04 binds preferentially to the free hLHbeta, (ii) LHP03 recognizes weakly the hCGbeta and oLHbeta in comparison to hLHbeta and (iii) LHP04 binds oLHbeta as well as hLHbeta but does not bind to hCGbeta and eLHbeta. The topographical relationship of epitopes recognized by LHP03 and monoclonal antibodies recognizing dimer specific epitopes on hLH allowed us to localize discontinuous antigenic sites that overlaps or are located outside the hHLbeta(43-52) region. Together, our results demonstrated that the hHLbeta(43-52) portion is accessible on both the free hLHbeta subunit and hLH whereas the COOH-terminal portion, hHLbeta(110-117), is probably buried at the alpha/beta interface of the hormone. PMID- 9397934 TI - The Hep G2 cell line in the study of growth hormone receptor/binding protein. AB - This study identifies specific, high affinity GH-receptors (GH-R) in human hepatoma Hep G2 cells. The binding characteristics of GH-R in the Hep G2 cells are similar to those of human liver membranes, such as the high specificity for hGH, the binding affinity (Ka = 1.7 +/- 0.5 x 10[9] M[-1]) and the molecular weight of the membrane bound GH-R (apparent 125,000 and 71,000). In addition, lower molecular weight forms (approximately 94,000 and approximately 58,000) were identified as GH-binding protein (GH-BP) in Hep G2 conditioned medium, or following incubation of Hep G2 cells, in the presence of 10 mM N-ethylmaleimide for 90 min at 30 degrees C; the latter are presumed to be shed by a proteolytic cleavage of the GH-R. Exposure of Hep G2 cells to physiologic concentrations of hGH resulted in a concentration-dependent increase in 3H-thymidine incorporation, up to 48.4 +/- 7.9% above control. In summary, the demonstration of specific, high affinity GH-R in Hep G2 cells, as well as shedding of GH-BP, suggest these cells may provide a homologous human system to study the receptor-effector interrelationship of hGH and to further our understanding of hepatocyte production of soluble GH-BP. PMID- 9397935 TI - Transcriptional and translational regulation of LH, prolactin and their testicular receptors by hCG and bromocriptine treatments in adult and neonatal rats. AB - Effects of altered gonadotropin and prolactin (PRL) secretion on luteinizing hormone (LH), PRL and their testicular receptors (R) were studied in neonatal and adult rats. Changes in gene expression were monitored by measurements of steady state mRNA levels. Five-day and 90-day-old male rats received a single s.c. injection of hCG (600 IU/kg), 1 mg/kg bromocriptine (BR) twice daily, or their combination. After 2 or 8 days, the responses of LH, PRL, their testicular R, and testosterone (T) were assessed, including measurements of the appropriate mRNA levels. Vehicle-treated age-matched animals served as controls. hCG suppressed serum LH in 2 days in adult rats from 0.85 +/- 0.16 to 0.04 +/- 0.01 microg/l, and in neonates from 0.59 +/- 0.29 to levels below 0.01 microg/l (p < 0.01 for both). This was accompanied at both ages by a 60% decrease in pituitary content of the LH beta-subunit mRNA (p < 0.01), but a decrease in the alpha-chain (40%, p < 0.05) occurred only in neonates. hCG increased serum PRL in adult rats in 8 days over 2-fold (p < 0.01); this did not occur in neonates. In neonates, BR increased the LH subunit mRNAs 2-fold in 8 days (p < 0.01) without a concomitant effect on serum LH; no BR effects on the LH parameters were seen in adult animals. BR decreased pituitary PRL protein and mRNA levels at both ages (p < 0.01-0.05), but serum PRL decreased only in the adults. The homologous down regulation of testicular LHR (near 100%) was accompanied in adults by a 30% decrease in LHR mRNA (p < 0.05). Also BR at this age decreased LHR binding (75% in 8 days, p < 0.01), but in this case no change occurred in the cognate mRNA. hCG and BR slightly up-regulated in adults PRLR binding, but only the 2-day effect of BR was accompanied by a 60% increase in PRLR mRNA (p < 0.05). In neonates, both hCG and BR increased testicular LHR and PRLR mRNA levels (p < 0.01 0.05). In adult animals, both hCG and BR suppressed testicular and serum T levels after 8 days (40-70%, p < 0.01-0.05); only BR was inhibitory to T by 8 days in the neonates (p < 0.05). In conclusion, the homologous and heterologous regulatory effects of hCG and BR on LH, PRL and their testicular R levels were only partly explained by changes in steady-state levels of the respective mRNAs. In general, the autoregulatory effects on LHR and PRLR appeared to affect steady state levels of cognate mRNAs, whereas heteroregulation predominately involved changes at the protein level. The responses of the neonatal pituitary-gonadal axis to hCG and/or BR differed greatly from those observed in the adult, indicating that the mechanisms involved in these regulatory events in adult animals are a result of gradual postnatal development. PMID- 9397936 TI - Catecholestrogen modulation of steroid production by rat luteal cells: mechanism of action. AB - This study investigated the mechanisms underlying 2-hydroxyestradiol (2-OHE2) effect on luteal steroidogenesis using serum-free cultures of mixed luteal cells from day 8 pseudopregnant rats. Initially, interactions between 2-OHE2 and LH or dibutyryl (db)cAMP on progesterone production were investigated. LH (250 ng/ml) and 2-OHE2 (2.5 microg/ml) had comparable effects on progesterone accumulation, while dbcAMP (5 mM) was more stimulatory. When applied together, 2-OHE2 did not synergize with LH or dbcAMP to further enhance progesterone accumulation. Furthermore, in time course experiments, the dose-dependent effect of 2-OHE2 was to reduce and eventually abolish the time-dependent increase in cAMP accumulation. In contrast LH stimulated cAMP accumulation at all times. Experiments in which cells were co-treated with 2-OHE2, 22-OH-cholesterol and cyanoketone, or with 2-OHE2 and 22-OH-cholesterol or pregnenolone indicated that 2-OHE2 not only had a stimulatory effect on the cholesterol side-chain cleavage and 3beta-hydroxysteroid dehydrogenase enzymes, but it also appeared to inhibit the 20alpha-hydroxysteroid dehydrogenase leading to a relative increase in progesterone accumulation. Experiments with hormone antagonists suggested that the actions of 2-OHE2 were not mediated by the estrogen, alpha- or beta adrenergic receptors. The results of this study support the concept of a physiological role for catecholestrogens in rat luteal steroidogenesis. PMID- 9397937 TI - Role of specific isoforms of protein kinase C in angiotensin II and lipoxygenase action in rat adrenal glomerulosa cells. AB - Evidence indicates that the lipoxygenase (LO) pathway of arachidonic acid is a key mediator of angiotensin II (AII)-induced aldosterone synthesis in adrenal glomerulosa cells. Although protein kinase C (PKC) may play a role in AII action, the precise PKC isoforms involved and whether LO products can activate PKC is not clear. We therefore evaluated the effect of AII and LO products such as 12- and 15-hydroxyeicosatetraenoic acids (HETEs) on PKC activation in isolated rat adrenal glomerulosa cells. PKC activity was measured by the phosphorylation of a PKC specific peptide while the PKC isoforms were identified by Western immunoblotting using antibodies that recognize the alpha, beta, gamma or epsilon isoforms of PKC. Treatment of the cells for 15 min with AII (10[-8]M) or the LO products 12- or 15-HETE caused a marked increase in PKC activity in membrane fractions with reciprocal decreases in the cytosolic PKC activity. Rat glomerulosa cells expressed only the alpha, and epsilon isoforms of PKC. AII increased membrane bound levels of both PKC-alpha and -epsilon (1.9- and 1.5 fold, respectively), whereas the LO products predominantly activated PKC-epsilon. Reciprocal decreases in immunoreactive cytosolic PKC levels were seen. AII induced aldosterone synthesis was blocked by H-7 and retinal as well as by a PKC specific pseudosubstrate inhibitor, PKC(19-36). These results suggest that AII and LO pathway-induced actions in the adrenal glomerulosa may be mediated by specific PKC isoforms. PMID- 9397938 TI - Regulation of uterine collagenase gene expression: interactions between serotonin and progesterone. AB - This report seeks to further define the requirements for the previously established induction of collagenase gene expression by serotonin and inhibition by progesterone in primary cultures of rat uterine smooth muscle cells. Detectable increases in collagenase production were observed after as little as 3 h exposure of cells to 5 microM serotonin, with maximal induction occurring after approximately 8 h of exposure. The apparent half-life of collagenase mRNA upon removal of serotonin was estimated to be approximately 12 h, and was not dependent on the duration of induction. Inhibition by either cycloheximide or progesterone showed similar half lives for collagenase mRNA, however a much shorter half-life (6 h) was obtained in the presence of actinomycin D. These experiments suggest that neither serotonin induction nor progesterone inhibition of collagenase synthesis represents a primary effect on collagenase gene transcription. Rather they appear to be secondary to changes that occur at one or more primary intermediate genes whose induction or decay must occur prior to changes in collagenase transcription. The progesterone receptor antagonist, RU 486, abrogates the ability of progesterone to inhibit serotonin-induced collagenase gene expression, indicating that the effects of progestins in this system likely are receptor-mediated. Finally, the present studies demonstrate that pretreatment of cells for times as long as 5 days with medroxyprogesterone in the absence of serotonin is unable to prevent subsequent serotonin-induced collagenase mRNA increases. These data suggest the possibility of a unique interaction between the molecular pathways of inducer and inhibitor, one in which serotonin may help mediate the progesterone-dependent repression of the levels of collagenase mRNA. PMID- 9397939 TI - The 5'-untranslated region of the IGF-I receptor gene modulates reporter gene expression by both pre- and post-transcriptional mechanisms. AB - The insulin-like growth factor I (IGF-I) receptor gene has a large, complex 5' untranslated region (UTR). In order to examine the role that this region plays in regulating IGF-I receptor expression, we isolated fragments of the human IGF-I receptor 5'-UTR and interposed them between the SV40 early promoter and the chloramphenicol acetyl transferase reporter gene. Fragments of the IGF-I receptor gene 5'-UTR were found to enhance reporter gene expression by increasing gene transcription. In addition, the increased transcription and mRNA levels were in excess of the increase in enzyme activity, providing indirect evidence that these fragments inhibit translation, consistent with predictions. PMID- 9397940 TI - Expression of insulin-like growth factor-I (IGF-I) receptor gene in rat brain and liver during development and in regenerating adult rat liver. AB - Insulin-like growth factor-I (IGF-I) is involved in the growth and development of liver and brain during fetal life by acting through specific plasma membrane receptors. In an attempt to determine how the changes in IGF-I receptor number are regulated during development, we have compared [125I]IGF-I binding to membrane fractions with the concentration of IGF-I receptor mRNA in rat liver and brain. IGF-I binding to liver membranes was 4.5 times higher in 20-day-old fetuses than in adult rats. After partial hepatectomy (70%) in adult rats a transient 2-fold increase of IGF-I binding to liver membranes was observed. In fetal and regenerating liver increases similar to those observed for IGF-I binding were observed in IGF-I receptor mRNA concentrations. In brain microsomes IGF-I binding was 3.5 times higher in 20-day-old fetuses than in adults. This difference reflects a similar change in the number of IGF-I receptors without modifications in the affinity of the receptor for the ligand. In contrast to the liver, no significant changes in the concentration of IGF-I receptor mRNA were observed in the developing brain when determined by hybridization solution, Northern blot or RNase protection analysis. These findings suggest that in the liver, the IGF-I receptor gene is regulated at pretranslational level during development and regeneration, while in brain it is preferentially regulated at translational or posttranslational level. PMID- 9397941 TI - Basic fibroblast growth factor induces luteinizing hormone receptor expression in the presence of insulin-like growth factor-I in ovarian granulosa cells. AB - Effect of basic fibroblast growth factor (bFGF) on the expression of receptors for luteinizing hormone (LH), a marker of differentiation, was studied using estrogen-primed rat ovarian granulosa cells in primary culture. bFGF had no effect by itself but dose-dependently induced expression of functional LH receptors in the presence of insulin-like growth factor-I (IGF-I). The effect of a combination of bFGF and IGF-I was delayed in onset and the magnitude of the response was smaller when compared to the action of follicle-stimulating hormone (FSH). Scatchard analysis revealed that dissociation constant (Kd) and number of LH receptors induced by bFGF and IGF-I were 0.47 nM and 6.48 fmol/10(6) cells, respectively. Unlike FSH, bFGF plus IGF-I did not cause an immediate increase in cAMP release, however, considerable amount of cAMP release was observed in cells incubated for 72 h with bFGF plus IGF-I. Indomethacin, an inhibitor of cyclooxygenase, attenuated both LH receptor expression and cAMP release induced by bFGF plus IGF-I but had little effect on the action of FSH. Finally, a combination of bFGF and IGF-I increased production of prostaglandin E2 in granulosa cells. These results indicate that bFGF is capable of inducing LH receptor in the presence of IGF-I by a mechanism involving production of prostaglandin E2. PMID- 9397942 TI - Characterization of estrogen receptor cDNAs from human uterus: identification of a novel PvuII polymorphism. AB - Using reverse transcription and the polymerase chain reaction, we have cloned estrogen receptor complementary DNAs from normal human uterine tissue. Restriction endonuclease analysis identified a polymorphic PvuII recognition site within half of the receptor cDNAs. Sequence analysis revealed a number of differences with the sequence previously reported for the ER cDNA isolated from MCF7 cells and confirmed that the codon for amino acid 400 was erroneously assigned as valine (GTG) rather than glycine (GGG). Sequencing also defined the nature of the PvuII polymorphism, with allele A coding for Glu22 and allele B (with an additional PvuII site) coding for Gln22. We demonstrate that both alleles of this receptor activate transcription of an estrogen-responsive gene to the same extent. This selective cloning method should have wide application in the investigation of naturally occurring cDNA variants from diseased tissues, such as breast cancer cell lines and primary tumor specimens. PMID- 9397943 TI - Recombinant murine tumor necrosis factor-alpha inhibits cholesterol side-chain cleavage cytochrome P450 and insulin-like growth factor-I gene expression in rat Leydig cells. AB - The purpose of the present study was to evaluate the effects of murine recombinant tumor necrosis factor-alpha (TNF-alpha) on rat Leydig cell function. In primary cultures of Leydig cells, we found that in the presence of hCG (10 ng/ml), testosterone levels were markedly elevated, 69.3 +/- 3.1 ng/10(6) cells/h (mean + SE). TNF-alpha in a concentration of 1 ng/ml markedly inhibited testosterone biosynthesis (a 69% reduction; p < 0.01) and 100 ng/ml of TNF-alpha almost completely inhibited testosterone formation (p < 0.001). TNF-alpha (10 ng/ml) inhibited hCG (0.1, 1 and 10 ng/ml)-induced testosterone formation by 63%, 67% and 61%, respectively. TNF-alpha (10 ng/ml) also markedly inhibited 8-bromo cAMP-induced testosterone formation from 76 +/- 9 ng/10(6) cells/h to 4.9 ng/10(6) cells/h. This indicates that the major effect of TNF-alpha is at steps beyond LH receptor site. To further evaluate the site(s) of action of TNF-alpha, we evaluated its effect on the conversion of precursor steroids to testosterone. We found that the addition of 20-hydroxy-cholesterol could not reverse inhibitory effects of TNF-alpha on hCG-induced testosterone formation. TNF-alpha had no effect on the conversions of pregnenolone, 17-OH-pregnenolone, DHEA and androstenedione to testosterone. This indicates that the major effect of TNF alpha is at the key steroidogenic enzyme, P450scc. We reported previously that human recombinant TNF-alpha had no effect on hCG-induced testosterone formation but did enhance the inhibitory effects of human recombinant IL-1beta. In the present study, we demonstrated that both murine TNF-alpha and human IL-1beta were potent inhibitors of hCG-induced testosterone formation. IL-1beta alone in concentrations of 0.1, 1 and 10 ng/ml inhibited testosterone formation by 45%, 62% and 91%, respectively, in the presence of TNF-alpha (10 ng/ml), IL-1beta in a concentration as low as 0.1 ng/ml completely blocked hCG-induced testosterone formation. We next evaluated the effect of TNF-alpha on P450scc gene expression. There was no constitutively expressed P450scc mRNA in Leydig cells after 24 h in culture. In response to hCG, there was a 33-fold increase in the P450scc mRNA level. Both TNF-alpha and IL-1beta inhibited hCG-induced expression of P450scc mRNA. Finally, the effect of TNF-alpha on IGF-I gene expression was investigated since IGF-I enhances Leydig cell androgen formation and IGF-I gene is expressed in high levels in Leydig cells. TNF-alpha inhibited both large (7.4 kb) and small species (0.8-1.2 kb) IGF-I mRNA levels in a dose-dependent manner. In conclusion, murine TNF-alpha is a potent inhibitor of Leydig cell function. TNF-alpha inhibited both P450scc and IGF-I mRNA gene expression. PMID- 9397944 TI - Role of the exon 11 of the insulin receptor gene on insulin binding identified by anti-peptide antibodies. AB - The insulin receptor exists in two isoforms differing by the absence (HIR-A) or presence (HIR-B) of 12 amino acids in the C-terminus of the alpha-subunit as a consequence of alternative splicing of exon 11. It was shown that the two isoforms exhibit different binding affinities for insulin, thus suggesting that the sequence encoded by exon 11 may be important for insulin binding. To further investigate this issue, we generated polyclonal antibodies against C-terminal peptides of the two HIR alpha-subunit variants. Herein, we characterized two antibodies, PA-11 and PA-12, directed against the C-terminus or the N-terminus of the sequence encoded by exon 11, respectively, and one (PA-13) directed against a sequence in the carboxy-terminal region of the alpha-subunit which is common to HIR-A and HIR-B. Antibodies were characterized for their ability to immunoprecipitate the receptor and to inhibit [125I]insulin binding to both isoforms. We found that PA-13 immunoprecipitates both the HIR-A and the HIR-B, PA 12 immunoprecipitates exclusively the HIR-B, and PA-11 fails to precipitate both isoforms. Interestingly, PA-12 inhibits specifically insulin to the HIR-B, whereas other PAs fail to affect insulin binding to either isoforms. Furthermore, PA-12 linearises the Scatchard plot of binding data, and retards the dissociation rate of insulin, thus suggesting that antibody affects cooperative interactions among binding sites. We conclude that the sequence encoded by exon 11 may play a role in modulating the binding of insulin to the receptor and negative cooperativity. PMID- 9397945 TI - Identification of a liver-specific promoter for the ovine growth hormone receptor. AB - Growth hormone (GH) receptor cDNA clones from several species are characterized by heterogeneity in the 5' untranslated region (5'UT). This has been attributed to different promoters directing the expression of the gene from exons encoding 5'UT's which are alternatively spliced onto a common splice acceptor 11 basepairs (bp) upstream of the initiating AUG on exon 2. The following study identifies exon 1A of the ovine (o) GH receptor gene, corresponding to the 5'UT of a developmentally regulated, liver-specific transcript. Exon 1A spans 206 bp at a position 17 kilobases (kb) upstream of exon 2. Sequencing of the 669 bp region 5' to the transcription initiation site (+1) reveals a TATA box at -31, a CCAAT box at -88, and putative binding sites for several transcription factors involved in liver-specific gene expression. Two repetitive sequence elements are located in the 5' and 3' flanking regions of exon 1A. Functional analysis of the 4.5 kb region upstream of exon 1A was performed by transfecting the human hepatoma cell line HuH7 with luciferase reporter gene constructs. Positive and negative regulatory regions are identified, with basal promoter activity within 473 bp of the transcription initiation site. A 47 bp region containing putative binding sites for the activated glucocorticoid receptor and C/EBP-like proteins, between 180 and -133, is essential for transcriptional activation. PMID- 9397946 TI - Human colon carcinoma cells (CaCo-2) synthesize IGF-II and express IGF-I receptors and IGF-II/M6P receptors. AB - The IGFs have been implicated in the development of the intestinal tract. We have studied the human colon carcinoma cell line CaCo-2 to gain more insight into the function of the IGFs in the gut. [125I]IGF-I and -II bound specifically to CaCo-2 cells as measured in competitive binding experiments. The existence of IGF-I receptors was further demonstrated by affinity crosslinking studies using DSS as the crosslinking agent. Western blotting of CaCo-2 cell extracts using an anti IGF-II/M6P receptor antiserum provided additional evidence for the expression of the IGF-II/M6P receptor. In addition, Northern blotting experiments showed specific IGF-I receptor and IGF-II/M6P receptor gene expression in CaCo-2 cells. An 11 kb band was visualized with a 614 bp PstI IGF-I receptor probe on autoradiographs. Hybridization with a 663 bp IGF-II/M6P receptor probe yielded a 9 kb RNA species. Analysis of CaCo-2 cell RNA using solution hybridization/RNase protection assays yielded two protected fragments, approximately 379 bases in length, with a 394 base IGF-I receptor riboprobe and a 250 base protected fragment with a 260 base IGF-II/M6P receptor riboprobe. In a subset of experiments a PstI 700 base fragment of the IGF-I cDNA and a 554 base SalI fragment of the IGF-II cDNA were used for hybridization: no hybridization was detected with the IGF-I probe. However, using the [32P]IGF-II probe bands at 6.0 and 5.0 kb were labeled in Northern blotting experiments. Analysis of CaCo-2 cell RNA using solution hybridization/RNase protection assays yielded a 289 base protected fragment and a faint 534 base species with a 556 base human IGF-II riboprobe. In addition, IGF-II immunoreactivity was measured in CaCo-2 cell conditioned medium using an IGF-binding protein blocked radioimmunoassay. CaCo-2 cell-conditioned medium contained 5-15 ng/ml IGF-II immunoreactivity. In conclusion, (1) CaCo-2 cells express both IGF-I receptor mRNA and IGF-II/M6P receptor mRNA and contain functional IGF-I receptor and IGF-II/M6P receptor protein. (2) CaCo-2 cells express IGF-II mRNA and secrete IGF-II immunoreactivity. We hypothesize that in human colon carcinoma cells IGF-II could act as an autocrine growth factor or alternatively could serve as a regulatory factor during differentiation. PMID- 9397947 TI - Rat gonadotropin-releasing hormone (GnRH) receptor: tissue expression and hormonal regulation of its mRNA. AB - The binding of gonadotropin-releasing hormone (GnRH) to its receptor in the anterior pituitary gland is the key molecular interaction regulating the reproductive process of mammals. Here, we report the isolation of a cDNA representing this receptor from rat anterior pituitary and the regulation of expression of its mRNA. The rat GnRH receptor cDNA was composed of 2909 nucleotides and encoded a protein containing 327 amino acids having a seven transmembrane topology. Northern blot analysis on RNA from rat pituitary, ovary and testis showed four different transcripts (5.0, 4.5, 2.5 and 1.3 kb) of which the 5.0 kb form was most abundant. The levels of expression of the transcripts were found to be highest in the pituitary followed by the ovary and the testis (about 40% and 5% compared to pituitary, respectively). Using the more sensitive reverse transcriptase/PCR technique, we also detected GnRH receptor mRNA in the adrenal and the hypothalamus. Measurement of pituitary GnRH receptor mRNA levels (the 5.0 kb form) during the estrous cycle showed the lowest levels at estrus (1.0-fold), a 2.2 +/- 0.57 (mean +/- SEM) -fold increase at diestrus I, a 3.5 +/- 0.41-fold increase at diestrus II, a 2.6 +/- 0.34-fold increase on the morning of proestrus, and a 1.9 +/- 0.25-fold on the afternoon of proestrus. Removal of the ovaries led to a 2.7 +/- 0.29-fold increase in GnRH receptor mRNA levels in the pituitary gland; treatment of ovariectomized rats with estrogen resulted in a significant decrease in GnRH receptor mRNA levels. Our studies demonstrate ovarian regulation of GnRH receptor mRNA expression in the anterior pituitary gland. PMID- 9397948 TI - Phorbol 12-myristate 13-acetate and 1,25-dihydroxyvitamin D3 regulate 1,25 dihydroxyvitamin D3 receptors synergistically in rat osteosarcoma cells. AB - In this study, we examined the effect of activation of protein kinase C (PKC) pathways on the regulation of 1,25-dihydroxyvitamin D receptors (VDR) in rat osteosarcoma (ROS) 17/2.8 cells. Activation of PKC with phorbol 12-myristate 13 acetate (PMA) resulted in a time- and dose-dependent increase in VDR expression in ROS cells. Treatment of ROS cells with 4alpha-phorbol 12,13-dedeconate, a PKC inactive phorbol ester, had no effect on VDR expression. Oleoyl acetyl glycerol (OAG), a synthetic diacylglycerol, stimulated VDR up-regulation in ROS cells. The PKC inhibitors (H-7, staurosporin, and sphingosine) all blocked PMA-mediated up regulation of VDR in a dose-dependent manner. We next examined the interaction of 1,25(OH)2D3 and PKC activation by PMA on the regulation of VDR in ROS cells. We found that PMA or 1,25(OH)2D3 treatment alone resulted in a 50 and 200% increase in VDR, respectively. PMA treatment alone resulted in a 50% increase in VDR protein and a marginal 20% increase in VDR mRNA. 1,25(OH)2D3 up-regulation of VDR was associated with a 2-fold increase in VDR mRNA. In contrast, co-treatment of ROS cells with PMA and 1,25(OH)2D3 resulted in a synergistic 10-fold induction of VDR mRNA and the appearance of a 7.2 kb VDR transcript. VDR protein was also synergistically up-regulated by combined PMA and 1,25(OH)2D3 treatment of ROS cells. Scatchard analysis demonstrated that the synergistic effect of PMA and 1,25(OH)2D3 on VDR protein expression was not associated with any change in the affinity of VDR for 1,25(OH)2D3. The synergistic effect of 1,25(OH)2D3 and PMA on VDR expression supports a link between PKC signal pathways and the function of VDR. PMID- 9397949 TI - Identification of the estrogen sensitive marker in human endometrial carcinoma RL95-2 cells. AB - Estrogen exerts a variety of biological effects on human reproductive tissues. However, little is understood about the estrogenic effect on human endometrial cells in vitro. This study was designed to investigate estrogen action on c-myc and c-fos oncogenes and lactoferrin gene expression in human endometrial carcinoma RL95-2 cells. The results indicate that estrogen can induce c-myc oncogene expression in 4 h. Neither c-fos nor the lactoferrin messenger was detectable, nor could they be induced by estrogen. Transfection with human estrogen receptor expression vector to the RL95-2 cells does not restore the estrogen responsiveness. In addition to estrogen, epidermal growth factor (EGF) and tumor promoter 12-O-tetradecanoylphorbol 13-acetate (TPA) can also induce c myc expression with no effect on c-fos or lactoferrin expression. Our data suggest that the c-myc oncogene in human endometrial carcinoma RL95-2 cells is the sensitive target gene for steroid hormone and growth factor action. PMID- 9397950 TI - A comparative study of the role of adenylate cyclase in the release of adrenocorticotropin from the ovine and rat anterior pituitary. AB - The interaction between corticotropin-releasing factor (CRF) and arginine vasopressin (AVP) is important in the regulation of adrenocorticotropin (ACTH) release from the anterior pituitary (AP). CRF exerts its effect on the AP by activating the adenylate cyclase (AC) complex whereas AVP increases the turnover of phosphatidylinositol. In the rat and in man, CRF is the most potent ACTH secretagogue whereas AVP alone is only a weak agonist. Since recent studies in the sheep indicate a reversal of this order of potency, these studies were undertaken to test the hypothesis that a functional alteration of the AC in the ovine corticotrope might limit the ability of CRF to release ACTH from these cells. When rat AP cells were incubated with CRF, a dose-dependent increase in AC activity was observed. This effect was potentiated either by AVP or PMA, although neither agent alone altered AC activity. In contrast, CRF alone, or in combination with AVP or PMA, did not increase AC activity in ovine AP cells. Both cholera toxin (CT) and pertussis toxin (PT) caused a dose-dependent release of ACTH from rat and ovine AP cells, but the amount of ACTH released from the ovine AP cells by both agents was relatively reduced. In the ovine cells, however, AVP acted synergistically with CT or PT to markedly increase the release of ACTH to levels which approached those obtained when the rat AP cells were exposed to CT or PT alone. Forskolin increased AC activity in AP cells of both species, but to a much lower extent in ovine cells than in the rat cells. However, when the ovine cells were exposed to AVP, the AC response to forskolin became similar to the response observed in the rat cells when incubated with forskolin alone. Forskolin also released significantly less ACTH from the ovine AP cells, but AVP also acted synergistically with forskolin to greatly enhance the amount of ACTH released from these cells. Finally, 8-bromo-cyclic AMP produced a similar release of ACTH from both ovine and rat AP cells. We conclude that: (1) the decreased ability of CRF to increase ACTH release from the ovine AP reflects a net decrease in AC activity and cannot be ascribed to an ovine corticotropic resistance to cAMP; (2) the decreased activity of the ovine corticotropic AC complex may in turn reflect functional alterations at the level of both the G proteins and the catalytic subunit; (3) since AVP causes protein kinase C substrate phosphorylation in the ovine AP, AVP may increase AC activity in this tissue by phosphorylating the G proteins and/or the catalytic subunit. PMID- 9397951 TI - Calcitriol attenuates the basal and vasoactive intestinal peptide-stimulated cAMP production in prolactin-secreting rat pituitary (GH4C1) cells. AB - A clonal strain of prolactin-producing rat pituitary tumour cells (GH4C1 cells) was used to study the effect of calcitriol on cyclic adenosine monophosphate (cAMP) production. Calcitriol (10 nM) attenuated both the basal and vasoactive intestinal peptide (VIP)-stimulated cAMP production after 2 days' pretreatment of the cells. The effect was detectable at 1 nM and maximal at about 10 nM. Calcitriol was at least 100 times more potent than calcidiol and 24 hydroxycalcidiol. Calcitriol (10 nM, 4 days) did not affect the specific binding of 125I-VIP, but attenuated the guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS) stimulated (100 microM) adenylyl cyclase activity by 25%. Calcitriol (10 nM, 4 days) also attenuated both the Mn2+ (1 mM) and the forskolin-stimulated (10 microM) adenylyl cyclase activity by 43 and 41%, respectively. In conclusion, these data suggest that calcitriol attenuates the basal and VIP-stimulated cAMP production by inhibiting the catalytic subunit of the adenylyl cyclase as well as the amount of the G protein Gs alpha. PMID- 9397952 TI - Pineal perfusion with calcium channel blockers inhibits differently daytime and nighttime melatonin production in rat. AB - In a previous study we have shown that the response of perifused pineal glands to calcium was different according to the circadian stage at which the glands were removed. This difference may be explained by circadian changes in calcium channel function. Therefore in the present study we documented the effects of calcium channel blockers in perifused rat pineal glands removed in the middle of the light and dark spans (7 and 19 HALO (hours after light onset), in a L/D 12:12 regimen). Moreover, we have studied the effect of calcium channel blockers on adrenergically stimulated pineal glands removed 7 HALO. Inorganic (Co2+ and Cd2+) and organic (nifedipine and diltiazem) calcium channel blockers at 10(-4) mol/l all significantly reduced melatonin production and this inhibition was more effective with the glands removed 7 HALO. In a concentration of 10(-)5 mol/l, only Cd2+ and diltiazem reduced melatonin production significantly in pineal glands removed 7 HALO. Verapamil at 10(-4) and 10(-5) mol/l showed no significant effect on melatonin production in glands removed both during the light and dark spans. Mn2+ at 10(-4) mol/l (but not at 10[-5] mol/l) appeared to stimulate melatonin production in glands removed both during the light and the dark (significant increase only with glands removed during the dark). Cobalt showed an immediate short inhibitory effect on both isoproterenol and norepinephrine stimulated melatonin release, whereas nifedipine showed a significant inhibition only on isoproterenol-stimulated melatonin release. These results strongly suggest a circadian stage dependence of the pineal gland response to some calcium channel blockers and the involvement of calcium in the release of melatonin from pinealocytes. PMID- 9397953 TI - Follicle-stimulating hormone receptor mRNA in the mouse ovary during post-natal development in the normal mouse and in the adult hypogonadal (hpg) mouse: structure of alternate transcripts. AB - The structure of RNA encoding the mouse ovarian follicle-stimulating hormone (FSH) receptor was studied during post-natal development and in the adult hypogonadal (hpg) mouse which lacks circulating gonadotrophins. Using reverse transcription and the polymerase chain reaction (PCR) four major transcripts of the FSH receptor were found in the normal adult ovary. The largest transcript was the expected size from the position of the PCR primers (on exons 1 and 10) and sequencing confirmed that it was derived from FSH receptor mRNA. The three other transcripts were also derived from FSH receptor mRNA but they contained deletions corresponding to one or more complete exons. Each transcript lacked exon 2 while exons 5 and/or 6 were lacking in the smaller species. All four transcripts were present in ovaries of hpg mice showing that expression of receptor mRNA and development of alternate splicing are not gonadotrophin-dependent. During development in the mouse full-length FSH receptor transcripts were not detected in the ovary until day 5 although shorter transcripts were present at days 1 and 3. Results confirm that the FSH receptor primary transcript undergoes alternate splicing in the ovary and that the pattern of splicing changes as the ovary develops, probably as a result of follicular development. PMID- 9397954 TI - Interactions between transport of triiodothyronine and tryptophan in JAR cells. AB - We studied the effect of a number of amino acids on uptake of L-triiodothyronine (T3) in the human choriocarcinoma cell line, JAR. Tryptophan inhibited saturable T3 uptake by about 57% without any significant effect on the non-saturable uptake. Michaelis constant (Km) for T3 uptake was 1.06 +/- 0.15 microM (n = 15) with the corresponding maximum velocity (Vmax) of 24.2 +/- 3.1 pmol/min/mg cellular protein. For tryptophan uptake the Km was 1.31 +/- 0.26 microM (n = 7) and Vmax was 166.4 +/- 35.7 pmol/min/mg protein. The kinetic parameters for both uptake processes were similar to those reported in normal placenta. Uptake of T3 was inhibited by tryptophan but not phenylalanine, but tryptophan uptake was inhibited both by T3 and phenylalanine. Inhibition of T3 uptake by tryptophan was dose dependent, with an inhibition constant (Ki) of 2.9 +/- 0.5 mM. Similarly, tryptophan uptake was inhibited by T3 and phenylalanine in a dose dependent way with Ki values of 4.9 +/- 0.5 microM and 15.6 +/- 4.8 microM respectively. Km for T3 uptake was significantly increased to 1.86 +/- 0.42 microM (n = 4) in the presence of 3 mM unlabelled tryptophan and, similarly, Km for tryptophan uptake was significantly increased to 9.91 +/- 2.57 microM (n = 3) in the presence of 5 microM unlabelled T3. Efflux of T3 was progressively inhibited by increasing concentrations of both ligands, i.e. was saturable. We conclude that there is mutual competitive inhibition between uptake systems for T3 and tryptophan in JAR cells, but the kinetic parameters of cross-inhibition of uptake by the substrates suggest that the carriers are distinct. T3 may be transported in JAR cells by at least two transport systems with differing substrate specificities. We also demonstrated the presence of a saturable membrane carrier mediating the efflux of T3 from the cells which was subject to trans-inhibition by T3 and tryptophan. PMID- 9397955 TI - Development of a bioassay for FSH using a recombinant human FSH receptor and a cAMP responsive luciferase reporter gene. AB - FSH exerts its actions primarily by increasing cAMP levels via a G protein-linked transmembrane receptor. We report the development of a bioassay for FSH using a cell line that stably expresses the human FSH receptor and a cAMP responsive human glycoprotein hormone alpha subunit luciferase reporter construct. Receptor activation by FSH was measured by changes in luciferase activity. The cell line was shown to express 1.6 x 10(4) receptors per cell which bound FSH with high affinity (Kd 2.76 x 10[-9] M). Human pituitary FSH caused a dose-dependent increase in cAMP (ED50, 190 mIU/ml) and luciferase (ED50, 31.5 mIU/ml) activity. The sensitivity of the bioassay was less than 0.6 mIU/well. Postmenopausal serum, rat, ovine and bovine FSH elicited a dose-dependent increase in luciferase activity. There was no significant stimulation by highly purified human LH or recombinant human TSH. This cell line should be useful in the determination of bioactive FSH and characterization of serum FSH inhibitors. PMID- 9397956 TI - Progesterone initiation of the human sperm acrosome reaction: the obligatory increase in intracellular calcium is independent of the chloride requirement. AB - The progesterone-initiated human sperm acrosome reaction (AR) requires a rise in intracellular Ca2+ ([Ca2+]i), extracellular Cl- and apparently increased Cl- flux through a unique steroid receptor/Cl- channel resembling but not identical to a GABA(A)/Cl- channel complex. The present study uses fura-2 loaded human sperm, GABA(A)/Cl- channel blockers (picrotoxin and pregnenolone sulfate) and Cl(-) containing and Cl(-)-deficient media to determine whether the progesterone mediated increase in [Ca2+]i is dependent on the Cl- requirement. There was no significant difference between the progesterone-mediated increases of [Ca2+]i obtained in Cl(-)-containing and Cl(-)-deficient media. Picrotoxin did not significantly inhibit the progesterone-mediated increase in [Ca2+]i, and pregnenolone sulfate increased [Ca2+]i to the same extent as progesterone. These results strongly suggest that the increase in [Ca2+]i essential to the AR is independent of the AR Cl- requirement and could be explained by the existence of two different sperm plasma membrane progesterone receptors. PMID- 9397957 TI - Gene transfer into Xenopus hepatocytes: transcriptional regulation by members of the nuclear receptor superfamily. AB - A procedure to culture Xenopus laevis hepatocytes that allows the cells in primary culture to be subjected to gene transfer experiments has been developed. The cultured cells continue to present tissue-specific markers such as expression of the albumin gene or estrogen-controlled vitellogenin gene expression, which are both restricted to liver. Two efficient and reproducible gene transfer procedures have been adapted to the Xenopus hepatocytes, namely lipofection and calcium phosphate-mediated precipitation. The transcription of transfected reporter genes controlled by estrogen-, glucocorticoid- or peroxisome proliferator-response elements was stimulated by endogenous or co-transfected receptor in a ligand-dependent manner. Furthermore, the expression of a reporter gene under the control of the entire promoter of the vitellogenin B1 gene mimicked the expression of the chromosomal vitellogenin gene with respect to basal and estrogen-induced activity. Thus, this culture-transfection system will prove very useful to study the regulation of genes expressed in the liver under the control of various hormones or xenobiotics. PMID- 9397958 TI - Functional analysis of an alternatively spliced estrogen receptor lacking exon 4 isolated from MCF-7 breast cancer cells and meningioma tissue. AB - An alternatively spliced mRNA coding for a variant estrogen receptor (ER) missing exon 4 (ERdelta4) was detected in the breast tumor cell line MCF7 and meningioma tissue by using the reversed transcriptase PCR technique. The trans-activational properties of this mutant ER were assessed in embryo carcinoma P19EC and human choriocarcinoma JEG3 cells by co-transfection of the ERdelta4 expression vector with an oxytocin promoter construct containing an estrogen-responsive element. ERdelta4 did not trans-activate the oxytocin promoter in either a hormone dependent or -independent manner. Co-transfection of ERdelta4 together with the wtER did not show any interference of ERdelta4 on the stimulation of the oxytocin promoter by the wtER. ERdelta4 was translated in vitro. Its capacity to bind estradiol, and the binding of the variant to a synthetic estrogen-responsive element were compared to those of the wild-type receptor. ERdelta4 did not bind to a synthetic estrogen-responsive element, nor did it bind estradiol. Hence, ERdelta4 appears to be a silent variant and we speculate that it is without any role in tumor progression. PMID- 9397959 TI - Arginine vasopressin (AVP) causes the reversible phosphorylation of the myristoylated alanine-rich C kinase substrate (MARCKS) protein in the ovine anterior pituitary: evidence that MARCKS phosphorylation is associated with adrenocorticotropin (ACTH) secretion. AB - We have recently shown that AVP causes a protein kinase C (PKC)-dependent increase in ACTH release and biosynthesis in ovine anterior pituitary cells. In these cells, AVP also causes the translocation of PKC from the cytosol to the cell membrane which is maximal at 5 min, but the intracellular events distal to protein kinase C activation that underlie ACTH secretion have not been well characterized to date. Since the MARCKS protein has been implicated in neurosecretion and is phosphorylated by PKC in synaptosomes, studies were carried out to determine whether AVP might cause MARCKS phosphorylation in the ovine anterior pituitary, and to determine whether this phenomenon might be temporally correlated with PKC translocation and the release of ACTH. When cytosolic fractions of rat brain, ovine anterior pituitary, and cultured ovine anterior pituitary cells were incubated with purified PKC, several proteins were phosphorylated including those in the region of 83-85 kDa. After precipitation of the proteins with 40% acetic acid, the 83-85 kDa phosphoproteins were selectively recovered in the acid soluble phase. Phosphopeptide maps of either the 83 or 85 kDa proteins were generated with Staphylococcus aureus V8 protease and revealed 13 and 9 kDa phosphopeptides, which are characteristic of the authentic MARCKS protein. An identical phosphopeptide map was also obtained when the MARCKS protein was selectively extracted from intact 32P-labeled anterior pituitary cells. MARCKS phosphorylation was markedly increased when ovine anterior pituitary cells were exposed to 1 microM phorbol 12-myristate 13-acetate (PMA). When the cells were exposed to 1 microM AVP, MARCKS phosphorylation increased at 15 s and reached the maximal plateau value at 30 s. MARCKS phosphorylation then started to diminish at 2 min, and baseline levels were attained by 10 min. In the same cells, AVP stimulated ACTH release in a biphasic manner - during the first 30 s, there resulted a rapid burst of ACTH secretion that was followed by a slower, but sustained rate of secretion. We conclude that: (1) AVP causes a rapid, and reversible, phosphorylation of the MARCKS protein in the ovine anterior pituitary; (2) since the AVP-induced increase in MARCKS phosphorylation occurs much earlier in these cells than does PKC trans-location, MARCKS phosphorylation may provide a more sensitive index of the onset of PKC activation than the translocation assay; (3) the close temporal association between MARCKS phosphorylation and the rapid early release of ACTH suggests that MARCKS phosphorylation may be involved in the initial intracellular events that underly exocytosis of the hormone. PMID- 9397960 TI - Identification and characterization of the glucagon receptor from adipose tissue. AB - 125I-glucagon was directly cross-linked to its receptor in isolated adipocyte plasma membranes using a UV irradiation procedure. This investigation resulted in identification of an adipocyte glucagon receptor complex of 62 kDa, present both in white and brown adipose tissues. The specificity of labeling was shown by interference of unlabeled hormone with incorporation of radioactive glucagon into 62 kDa species. Treatment of adipose plasma membranes with N-glycanase resulted in appearance of intermediate species, indicating that the glucagon receptor is modified with several N-linked oligosaccharide chains similarly to the hepatic glucagon receptor. Peptide mapping of the affinity labeled adipose membranes with Staphylococcus aureus V8 protease generated three distinct receptor fragments identical to that of the hepatic receptor. Overall, the biochemical characterization of the rat adipocyte glucagon receptor indicates that it closely resembles the hepatic glucagon receptor. PMID- 9397961 TI - Different cells and cell lines produce factors that modulate Sertoli cell function. AB - Peritubular myoid cells derived from immature rat testes produce factors that modulate Sertoli cell function (P-Mod-S). The secretion of these factors is controlled in part by androgens. Cultured prostatic stromal cells strongly resemble peritubular myoid cells and produce mediators with similar activity. Here we investigated whether myoid cell lines can be used as a source of P-Mod-S like factors. Rat kidney fibroblast (NRK) and mouse fibroblast (3T3) cell lines were used as non-myoid controls. Surprisingly, serum-free media conditioned by all cell lines studied modulated Sertoli cell function in a similar fashion as media conditioned by peritubular cells (PTCM) or stromal cells (STCM). Using Sertoli cell transferrin secretion as an endpoint for P-Mod-S-like activity, the nature of the active principles involved was further explored. The observed activity could not be explained by residual contamination with fetal calf serum. Moreover, the effects of the conditioned media could not be mimicked by classical growth factors (IGF-I, bFGF, EGF, TGF-beta, NGF, PDGF-BB) added singly or in combination with submaximally effective concentrations of PTCM. Finally, the possibility that conditioned media might indirectly enhance Sertoli cell function by promoting the production or deposition of extracellular matrix elements was made unlikely by the demonstration that the observed effects were not mimicked by Matrigel and were unaffected when Sertoli cells were seeded on Matrigel. Superdex 75 chromatography after analytical reversed-phase chromatography indicates that the factors from different origin have a similar size (45-50 kDa). It is concluded that mediators with P-Mod-S-like activity are produced by various cells and cell lines both with and without smooth muscle cell characteristics. Whether the active principles involved are really identical requires further investigation. PMID- 9397962 TI - Processing of proenkephalin in bovine chromaffin cells occurs in two phases. AB - The processing of proenkephalin was studied in primary cultures of bovine adrenal medullary chromaffin cells by pulse-chase radiolabeling, immunopurification of proenkephalin and derivative peptides and quantitation following gel electrophoresis and Western blotting. Proenkephalin was processed with a t1/2 of approximately 1.1 h. Processing of proenkephalin-derived peptides of 15-25 kDa was essentially complete by 1 h. Treatment of chromaffin cells with brefeldin A to block the intracellular transport of proteins or with ammonium chloride to neutralize acidic intracellular compartments had only minor effects on the initial processing of proenkephalin. In contrast, both of these agents prevented a second, slower phase of proenkephalin processing. These studies suggest that proteolytic processing of proenkephalin in bovine adrenal medullary cells starts before transport to the trans-Golgi network and packaging into the chromaffin granules. A second phase of processing that requires an acidic environment occurs in or distal to the trans-Golgi network. PMID- 9397963 TI - Thyroxine control of pancreatic amylase gene expression: modulation of PTF1 binding activity. AB - The role of pancreas specific transcription factor (PTF1) in thyroxine (T4) modulation of amylase gene expression in suckling rats was evaluated. Electrophoretic mobility shift assay (EMSA) was used to determine the PTF1 binding activity by the amount of a synthetic oligonucleotide containing the amylase enhancer sequence bound by nuclear protein extracts. Nuclear protein from rat pancreata showed a developmental increase of PTFI activity correlated with age. To study the action of T4, pups were made hyperthyroid by T4 injection and hypothyroid by feeding propylthiouracil (PTU) to the lactating dams. EMSA of nuclear proteins isolated from these groups showed an increase in PTF1 binding activity in the T4 group and a decrease in the PTU group. Concomitantly, T4 increased, while PTU decreased both amylase enzyme and mRNA concentrations. T4 replacement reversed the effect of PTU on PTF1 binding, amylase enzyme activity and mRNA levels. To examine the age dependence of T4 effects, T4 was injected to pups for 5 days prior to killing at the age of 15, and 25 days. T4 was effective when given at an earlier age (15 days) but not at a later stage (25 days) in increasing amylase activity and amylase mRNA levels. Nuclear proteins isolated from pancreata of these groups showed an increase in PTF1 binding activity in the T4-treated 15-day-olds but not in the 25-day-olds in comparison to their corresponding age matched littermates. These results suggest that PTF1 is an important intermediary in T4 modulation of amylase gene expression during ontogeny of the rat exocrine pancreas. PMID- 9397964 TI - Dexamethasone regulation of parathyroid hormone-related protein (PTHrP) expression in a squamous cancer cell line. AB - Dexamethasone regulation of PTHrP expression has been studied in an epidermal squamous cancer cell line COLO 16, which secretes immunoreactive PTHrP into conditioned medium. Dexamethasone was found to suppress PTHrP expression in a time- and dose-dependent manner, which was reversible upon removal of dexamethasone. The half-maximal effective concentration of dexamethasone was 1 nM and an effect of dexamethasone on PTHrP mRNA was first observed after 2 h of treatment, with maximal inhibition by 6 h. Dexamethasone action on PTHrP expression was steroid specific since progestin, 5alpha-dihydroxytestosterone and oestrogen did not regulate PTHrP expression in COLO 16 cells. The gluocorticoid/progesterone receptor antagonist RU486 inhibited the dexamethasone effect, indicating glucocorticoid receptor-mediated regulation of PTHrP expression. The half-life of PTHrP mRNA in COLO 16 cells was approximately 120 min and was not altered by treatment of cells with dexamethasone. Nuclear run-on assays revealed that dexamethasone reduced PTHrP gene transcription in COLO 16 cells. Transient transfection assays with a series of reporter gene constructs encompassing 3.5 kb of the 5' end of the PTHrP gene failed to identify a region of the gene responsible for glucocorticoid down-regulation. PCR of reverse transcribed RNA from COLO 16 cells revealed that dexamethasone down-regulated transcripts driven from all three promoters (i.e., the TATA promoters 5' to exons I and IV and the GC-rich promoter 5' to exon III) of the human PTHrP gene. PMID- 9397965 TI - Cytokine-mediated regulation of rat ovarian function: interleukin-1 inhibits plasminogen activator activity through the induction of plasminogen activator inhibitor-1 (PAI-1). AB - Intraovarian IL-1 has recently been implicated as a mediator in the ovulatory process. Since PA activation is an established component of the ovulatory cascade, consideration was given in this report to the possibility that IL-1 may modulate ovarian PA economy. Whole ovarian dispersates from immature rats (25-27 days-old) were cultured under serum-free conditions for 48 h in the absence or presence of IL-1beta. Cellular PA activity was measured by plasminogen-dependent cleavage of 14C-labeled globin. Cells grown in the absence of IL-1 exhibited appreciable PA activity, as assessed by the cleavage of 0.074 +/- 0.026 mg [14C] globin/5 x 10(5) cells (mean +/- SD). Exposure to IL-1 (10 ng/ml) led to a 30% reduction in cell-associated PA activity (p < 0.001). The IL-1-mediated inhibition occurred concurrently with a 10-fold increase in the ability of the corresponding conditioned media to inhibit exogenous urokinase activity. At similar cell densities of 5 x 10(5) cells/well, isolated cultures of theca and granulosa cells exhibited comparable PA activity in the absence of IL-1. However, only theca cells responded to IL-1 with inhibition of plasminogen activation and enhancement of urokinase inhibitory activity. Granulosa cells in turn failed to respond to IL-1. Both the inhibition of PA activity and the increase in urokinase inhibitory activity proved cell-density- and IL-1 dose-dependent. The IL-1 induced inhibition of urokinase was abolished by the administration of a polyclonal anti-rat PAI-1 IgG. Both effects of IL-1 were counteracted in a dose dependent fashion by the soluble IL-1 receptor (which specifically complexes with IL-1), and by a highly-specific IL-1 receptor antagonist suggesting that the IL-1 effects are receptor-mediated. The present observations indicate that ovarian PA activity is subject to inhibition by IL-1 probably by way of PAI-1 of theca interstitial origin. Inasmuch as IL-1 may be involved in initiating and maintaining the preovulatory cascade, the periovulatory activation of plasminogen must be accomplished by agents other than IL-1. PMID- 9397966 TI - Alteration of basal release of anterior pituitary hormones by pretreatment of primary cultured cells with trypsin. AB - The anterior pituitary (AP) gland secretes 6 different hormones. Prolactin (PRL) is secreted at a relatively high level without stimulation by the hypothalamus, while secretion of the others requires the action of stimulatory factors from the hypothalamus. In order to gain an insight into the mechanism underlying the different spontaneous release patterns of these hormones, we investigated their spontaneous release rate after pretreating rat anterior pituitary cells with trypsin. Rat AP cells were cultured on Cytodex microcarrier beads for 4 days and were then superfused with either control medium or medium containing trypsin (0.25%) for 5 min. The subsequent release rates of the AP hormones were monitored. The basal release of PRL was severely reduced to almost undetectable level and began to recover 120 min after the trypsin-pretreatment. Full recovery was attained over the next 100 min and was delayed by treatment with a protein synthesis inhibitor, cycloheximide (7 microM). In the trypsin-pretreated cells, basal release of PRL and growth hormone (GH) was severely reduced, while that of thyroid stimulating hormone (TSH) and adrenocorticotropic hormone (ACTH) was enhanced and luteinizing hormone (LH) and follicle stimulating hormone (FSH) was not markedly affected by the treatment, suggesting that the suppression of PRL release was not caused by nonspecific damage to the cells. Since trypsin does not readily enter cells, the altered secretion of AP hormones seems to be the result of restricted digestion of the external components of the cells. On the bases of these observations, we predicted that the mechanism of spontaneous release of hormones involves trypsin sensitive proteins (TSMP) on the plasma membranes of the anterior pituitary cells. PMID- 9397967 TI - Growth hormone (GH) and insulin-like growth factor-I (IGF-I) treatment of the GH deficient dwarf rat: differential effects on IGF-I transcription start site expression in hepatic and extrahepatic tissues and lack of effect on type I IGF receptor mRNA expression. AB - The rat IGF-I gene consists of six exons, with exons 3 and 4 forming a 'core' mature IGF-I coding region to which alternate 5' and 3' regions are spliced. Transcription occurs from four dispersed start sites (ss) approximately 382 (ss 1), approximately 343 (ss 2), approximately 245 (ss 3) and approximately 30-40 (ss 4) basepairs (bp) from the 3' end of exon 1, and from a region 50-70 bp from the 3' end of exon 2. The expression of ss mRNAs displays tissue-specific and ontogenic regulation. Alternate splicing of exon 5 produces E-peptide coding domain variants (Ea and Eb mRNAs), with the Eb form found predominantly in the liver. The regulation of IGF-I mRNA expression by GH and IGF-I in the GH deficient dwarf (dw/dw) rat was investigated using antisense RNA probes in a solution hybridization RNase protection assay to detect leader exon and E domain variant mRNAs. GH treatment of dw/dw and normal Lewis rats increased the expression of all liver leader exon ss and E domain variants coordinately (1.6 1.9-fold increase, p < 0.01), although the increase observed in Eb transcripts was significantly higher in the dw/dw compared to the normal rat (p < 0.05). In kidney, GH treatment significantly increased exon 1 ss 3 and ss 4 transcripts by approximately 40% (p < 0.05). The expression of the other start sites was not affected by GH, suggesting that transcription factors may regulate start site usage independently. GH treatment was associated with a significant increase in IGF-I mRNA expression in skeletal muscle (p < 0.05) but not cardiac muscle or spleen. IGF-I treatment was associated with minor (approximately 20%) but significant (p < 0.05) reductions in IGF-I mRNA expression in the liver and kidney of dw/dw rats, suggesting that IGF-I can suppress IGF-I mRNA expression. IGF-I treatment did not affect IGF-I mRNA expression in cardiac and skeletal muscle of dw/dw rats. IGF-I receptor mRNA was detected in extrahepatic tissues only, and was not affected by either GH or IGF-I treatment. In summary, start site-specific regulation by GH was observed in kidney. GH increased IGF-I mRNA expression in muscle, kidney and liver, but had no effect in heart or spleen in the dw/dw rat. Our data suggest that systemic IGF-I can feedback on hepatic and renal IGF-I mRNA expression in the GH-deficient state. PMID- 9397968 TI - Transforming growth factor-beta1 regulates steady-state PTH/PTHrP receptor mRNA levels and PTHrP binding in ROS 17/2.8 osteosarcoma cells. AB - The effect of transforming growth factor beta1 (TGF-beta1) on the expression of mRNA for the parathyroid hormone receptor and binding of iodinated parathyroid hormone-related protein in ROS 17/2.8 osteosarcoma cells was evaluated. TGF-beta1 stimulated a 2-7-fold increase in steady state mRNA levels for the parathyroid hormone receptor at a maximal dose of 5 ng/ml, with increased levels of expression at 6 h of TGF-beta1-incubation, and peak levels at 8-24 h. Receptor binding studies revealed a significant increase in PTHrP-specific binding with TGF-beta1 doses as low as 0.5 ng/ml and a 55% increase in numbers of receptors with no alteration in binding affinity with 5.0 ng/ml TGF-beta1. Time course studies indicated that receptor binding was increased at 24 h with peak levels reached at 48 h of treatment. PTH-stimulated cAMP levels were significantly increased in ROS 17/2.8 cells treated with TGF-beta1 (0.5 ng/ml) for 48 h. These data indicate that TGF-beta1 upregulates steady-state mRNA, ligand binding and PTH/PTHrP receptor signaling in rat osteosarcoma cells. The effects of TGF-beta1 on bone may be attributed in part to regulation of the PTH/PTHrP receptor at the molecular level. PMID- 9397969 TI - Multiple levels of control of insulin-like growth factor gene expression. PMID- 9397970 TI - Antisense oligonucleotide strategies in physiology. AB - Antisense oligonucleotides can inhibit gene expression in living cells by binding to complementary sequences of DNA, RNA or mRNA. The mechanisms include inhibition of RNA synthesis, RNA splicing, mRNA export, binding of initiation factors, assembly of ribosome subunits and of sliding of the ribosome along the mRNA coding sequence. The most efficient antisense oligonucleotides also activate RNAse H, an ubiquitous enzyme that cleaves the mRNA at sites of mRNA/oligonucleotide duplex formation. A staggering number of oligonucleotide modifications have been proposed to retard degradation by nucleases, enhance cellular uptake, increase binding to the target sequence, and minimize non specific binding to related nucleic acid sequences. Phosphorothioates are the most popular oligonucleotides used in cell culture and in vivo, although sequence non-specificity remains an underreported problem. Recently developed chimeras between methylphosphonates and phosphodiester oligonucleotides appear to combine the advantages of water solubility, nuclease resistance, enhanced cellular uptake, activation of RNAse H, and high sequence selectivity. Antigene oligonucleotides are also promising, because they can inhibit gene expression by triple helix formation with DNA or by binding to one of the DNA strands. They have so far been little used in physiological studies. Cost is still a prohibitive factor, especially for suppressing the expression of a hormone or hormone receptor gene in rats, for example. However, patch-clamp dialysis of single cells or nuclear microinjections in culture, exposure of cultures to extracellular oligonucleotides, and intra-cerebral microinjections of oligonucleotides are feasible and highly rewarding approaches in physiology. PMID- 9397971 TI - A constitutively active form of CREB can activate expression of the rat prolactin promoter in non-pituitary cells. AB - The pituitary cell-specific transcription factor Pit-1 has been show to trans activate expression of the prolactin (PRL) promoter in non-pituitary cells. However, the cyclic AMP response element (CRE)-binding protein CREB is known to play a major role in cell-specific expression of hepatocyte-specific genes. Since the PRL promoter contains an asymmetrical form of a cyclic AMP response element (termed the CLE), we investigated whether CREB could also induce PRL promoter activity in non-pituitary cells. Transient expression in rat glial C6 cells of a constitutively active CREB-VP16 fusion protein strongly trans-activated expression of a co-transfected rat PRL promoter construct, (-187)PRL-CAT. Analysis by 5'-deletion showed that this response requires PRL promoter sequences between positions -113/-75. CREB-VP16 did not stimulate expression in C6 cells of any of three control promoter-CAT constructs, implying that the strong response of the PRL promoter to activated CREB is both promoter-specific, and is not due to non-specific transcriptional effects of the potent VP16 moiety of CREB-VP16. Surprisingly, mutations in the CLE only slightly reduced activation by CREB-VP16 of construct (-204)PRL-CAT, implying that the major action of CREB-VP16 on the PRL promoter does not involve a direct interaction with the CLE. CREB-VP16 stimulated PRL-CAT activity in C6 cells as strongly as, and synergistically with, Pit-1. These results imply that CREB can strongly and specifically activate expression of the PRL promoter in non-pituitary cells, via a mechanism different from that employed by Pit-1. PMID- 9397972 TI - Cytokines modulate type I iodothyronine deiodinase mRNA levels and enzyme activity in FRTL-5 rat thyroid cells. AB - Tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), and interferon-gamma (INF-gamma) have inhibitory effects on thyroid function both in vivo and in vitro. We have studied the effects of these cytokines on type I 5' deiodinase (5'-DI) mRNA expression and enzyme activity in FRTL-5 cells maintained in standard cell culture medium containing 0 (5H) or 2 mIU/ml bovine TSH (6H). Northern blots were hybridized with 5'-DI cDNA. 5'-DI mRNA levels were reduced to 20% of control values after treating cells with 100 ng/ml TNF-alpha in 6H for 2 days while the corresponding enzyme activity was reduced 50%. Other cytokines, including IL-1beta and interferon-gamma, also significantly inhibited expression of 5'-DI in FRTL-5 cells grown in 6H medium. Because the majority of circulating T3 in the rat is secreted by the thyroid gland, the highly significant decline in the serum T3/T4 ratio following in vivo administration of cytokines may be due to their direct inhibitory effect on thyroidal 5'-DI expression. PMID- 9397973 TI - Hepatitis C virus infection and type II cryoglobulinemia: an immunological perspective. PMID- 9397974 TI - Blunted responsiveness of the neuronal activation marker Fos in brainstem cardiovascular nuclei of cirrhotic rats. AB - Cardiovascular function in cirrhosis is deranged, with indirect evidence of abnormal central cardiovascular regulation. We aimed to elucidate the role of brainstem cardiovascular nuclei in hemodynamic regulation by examining the protein product, Fos, of the immediate-early gene c-fos, in cirrhotic rats. Cirrhosis was induced by chronic bile duct ligation (BDL) of 25-days duration, while controls underwent a sham operation. To examine the effects of jaundice per se in the absence of cirrhosis, a third group of 5-day BDL rats was also studied. All rats were anesthetized with pentobarbital, and catheters were inserted to measure baseline blood pressure and heart rate. Separate groups were then subjected to volume manipulation by a hypotensive hemorrhage or isotonic saline infusion, or no challenge. Ninety minutes after the volume manipulation, the animals were killed and the medulla sectioned and stained for Fos by immunohistochemisty. The nucleus tractus solitarius (NTS) of the sham-operated unchallenged rats showed scant Fos immunoreactivity (27.8 +/- 3.3 cells), but both hemorrhage and volume infusion significantly increased Fos staining (86.0 +/ 3.7 and 95.2 +/- 8.5, respectively). In contrast, the unchallenged cirrhotic rats showed markedly increased Fos in the NTS (154.6 +/- 27.0), but neither hemorrhage nor volume infusion significantly changed the amount of Fos staining. Fos staining in the ventrolateral medulla (VLM) followed a similar pattern with low staining in the unchallenged sham rats and increased staining in the other groups, but no differences between the unchallenged and the volume-manipulated cirrhotic groups. The 5-day BDL jaundiced rats showed no baseline increase in Fos staining, nor any significant increase after hemorrhage. These results showing baseline activation of central neuronal regions responsible for blood pressure homeostasis, but completely blunted responsiveness in cirrhotic rats, confirm a central origin of disordered cardiovascular regulation. The presence of jaundice may also contribute to the central cardiovascular hyporesponsiveness. PMID- 9397975 TI - The effects of low dietary levels of polyunsaturates on alcohol-induced liver disease in rhesus monkeys. AB - Rhesus monkeys that were maintained on a diet containing low, yet adequate, amounts of vitamins C and E and in which linoleate and linolenate represented 1.4% and 0.08% of the total caloric intake, respectively, developed liver fibrosis after consuming alcohol (mean, 2.6 g kg(-1) d[-1]) over a period of 3 years. In the liver, several polyunsaturated fatty acids including 18:2n6, 20:4n6, and 22:6n3 decreased compared with dietary controls, and similar findings were also observed in plasma lipoproteins and erythrocytes. The amount of alcohol consumed correlated positively with plasma lipid peroxidation products, 4 hydroxynonenal (4-HNE) and 8-isoprostane F2alpha, and negatively with 20:4n6 and 22:6n3 levels. These findings imply that alcoholics who also have a marginal intake of essential fatty acids and antioxidants in their diets may be at an increased risk of developing liver disease. PMID- 9397976 TI - Clinical, histological, and virological features of hepatitis C virus carriers with persistently normal or abnormal alanine transaminase levels. AB - This study was aimed to evaluate demographic, clinical, histological, and virological characteristics of 46 hepatitis C virus (HCV) carriers with persistently normal alanine transaminase (ALT) levels and to compare the results with those obtained in a group of 52 HCV-RNA-positive patients with elevated ALT levels. Subjects with normal ALT were more often females (P < .001), were more likely to be asymptomatic (P < .001), and have a lower incidence of risk factors for HCV transmission (P < .01). All patients with normal ALT had significant histological liver damage. The mean grading and staging did not differ between patients with normal and those with raised ALT concentrations. Moderate to severe hepatitis was more frequently found among subjects with normal than with elevated ALT. HCV genotype 2a was far more common in subjects with normal (43%) than with abnormal ALT levels (6%; P < .002), genotype 1b being more frequent in these latter (50% vs. 17%; P < .001). Patients with normal ALT levels had similar serum HCV-RNA titers than subjects with raised ALT. Neither HCV genotype distribution nor viral load correlated with the severity of liver damage. We conclude that significant liver disease may occur irrespective of clinical symptoms, ALT levels, HCV genotypes, and viral load. PMID- 9397977 TI - Enhanced apoptosis relates to bile duct loss in primary biliary cirrhosis. AB - Primary biliary cirrhosis (PBC) is characterized by an immune-mediated destruction of intrahepatic small bile ducts. Apoptosis, a unique pattern of cell death, has been suggested to be responsible for the biliary destruction in PBC. To address this issue, we attempted to detect the apoptosis of biliary epithelial cells by in situ nick-end labeling and by the expression of apoptosis-related proteins using immunohistochemistry in patients with various hepatobiliary diseases, including PBC. The data was noteworthy for several reasons. First, apoptosis was occasionally detected on biliary cells in all liver specimens; however, the positive rate was high in PBC and relatively low in other livers. Strong expression of CD95 was frequently observed in the epithelial cells of the injured bile ducts of PBC, which accompanied high intensity CD95 ligand expressing mononuclear cells. Perforin and granzyme B immunoreactivities were occasionally found on the bile ducts in control liver diseases as well as PBC, but granzyme B-positive biliary cells were prominent in PBC. In contrast, Lewis Y expression, as detected using BM-1 antibody, was consistently present in the injured bile ducts of PBC. These data suggest that apoptosis, via the perforin/granzyme B pathway, may be associated with the degrading fraction of cell cycle regulation in the small-sized biliary tree under physiological and pathological liver conditions. Moreover, enhanced apoptosis, mediated by CD95/CD95 ligand interaction, may contribute to the bile duct injury and loss observed in PBC. PMID- 9397978 TI - Infection complicating percutaneous liver biopsy in liver transplant recipients. AB - There is controversy about the frequency of and risk factors for infectious complications of percutaneous liver biopsy in liver transplant recipients. The aim of this study was to identify the incidence and nature of complications associated with liver biopsy after orthotopic liver transplantation (OLT), with particular emphasis on infection. The medical records of all patients undergoing OLT between January 1990 and August 1994 were reviewed retrospectively to identify complications requiring hospitalization within one week of percutaneous liver biopsy. The nature and severity of complications were recorded and possible risk factors for infectious complications were examined. One hundred ninety-eight patients underwent 1,136 percutaneous liver biopsies. There were eleven complications (0.96%), including as follows: 7 infections, 3 bleeding episodes, and 1 vasovagal reaction. Infections after percutaneous liver biopsy included fever and bacteremia (n = 6), and fever without bacteremia (n = 1). All infections developed only in patients with underlying biliary tract abnormalities; the frequency of infection was higher (9.8%) in patients with choledochojejunostomy when compared with those with choledochocholedochostomy (1.4%). Bacteremia was more likely caused by skin flora in patients with choledochocholedochostomy (CDC) and by enteric bacteria in patients with choledochojejunostomy (CDJ). All infections were treated successfully with parenteral antibiotics. We conclude that biliary tract abnormalities are the primary risk factors for infection after percutaneous liver biopsy, although the risk is higher in patients with CDJ than with CDC. These data support the use of antibiotic prophylaxis before percutaneous liver biopsy in OLT recipients with biliary tract abnormalities. PMID- 9397979 TI - Clinical efficacy of lactulose in cirrhotic patients with and without subclinical hepatic encephalopathy. AB - Seventy-five cirrhotic patients with hyperammonemia in the past or at the time of the study were randomly divided into two groups (treated with lactulose or nontreated) in 14 hospitals in Japan. Thirty-six cirrhotic patients were diagnosed as having subclinical hepatic encephalopathy (SHE), and 39 were diagnosed as non-SHE. SHE was diagnosed when the results of all three of the quantitative psychometric tests used (number connection test, and symbol digit and block design tests of the Wechsler adult intelligence scale [revised]) were abnormal as compared with age-matched normal values. The mean number of abnormal psychometric test results and the prevalence of SHE were used for a quantitative evaluation of the efficacy of the lactulose treatment. Twenty-two of the SHE patients were treated with lactulose (45 mL/d) for 8 weeks, and the other 14 SHE patients did not receive lactulose. In the SHE patients administered lactulose, the results of the quantitative psychometric evaluation were significantly improved at 4 and 8 weeks after the beginning of the lactulose administration. The SHE had disappeared in 10 (50%) of the 20 treated patients at week 8, but it persisted in 11 (85%) of the untreated 13 patients. We concluded that lactulose treatment in cirrhotic patients with SHE is effective with respect to psychometric tests. PMID- 9397980 TI - Large cell change (liver cell dysplasia) and hepatocellular carcinoma in cirrhosis: matched case-control study, pathological analysis, and pathogenetic hypothesis. AB - Large cell change (LCC), characterized by cellular enlargement, nuclear pleomorphism and hyperchromasia, and multinucleation of hepatocytes, is a common lesion in cirrhotic livers, but its nature, significance, and pathogenesis remain uncertain. Therefore, we assessed the prognostic value of LCC as a marker of subsequent hepatocellular carcinoma (HCC) through a case-control study that compared pretransplant liver biopsy specimens from 37 cirrhotic liver transplant recipients with HCC to specimens from a control group of recipients without HCC, matched for sex, age (+/-5 years), and cause of cirrhosis. LCC was identified in 16 (43%) of the study and 7 (19%) of the control group biopsy specimens. By matched-pair analysis, LCC conveyed a moderately increased risk of later HCC with an estimated odds ratio of 3.3 (95% CI, 1.2-15; P = .038). However, a pathology review of 45 HCCs showed adjoining LCC in only 12 (27%) and did not suggest a morphological transition or a histogenetic association between the two lesions. LCC hepatocytes displayed a low proliferative rate by Ki-67 or proliferating cell nuclear antigen immunostaining (labeling indices of 0.27 and 0.73) but showed a greater degree of apoptosis than normal hepatocytes (labeling indices of 1.9 and 0.23; P = .03) To reconcile these findings, we propose that LCC derives from derangements in the hepatocyte's normal process of polyploidization. Such derangements, possibly caused by chronic inflammation-induced DNA damage, could yield a population of enlarged liver cells with nuclear atypia and pleomorphism, frequent binuclearity, and minimal proliferation. According to this hypothesis, LCC would be a habitual feature of cirrhosis and a regular accompaniment of HCC but would not represent a direct malignant precursor. PMID- 9397981 TI - Indomethacin normalizes intracranial pressure in acute liver failure: a twenty three-year-old woman treated with indomethacin. AB - In patients with acute liver failure, cerebral herniation is a common cause of death. The present study reports the effect of indomethacin on four occasions of intracranial hypertension, in a 23-year old previously healthy woman with severe acetaminophen poisoning. During each episode of intracranial hypertension, the patient was treated with 25 mg of indomethacin, and each time the intracranial pressure normalized. We recommend further controlled studies to determine the exact effect of indomethacin on cerebral blood flow and metabolism before it is recommended for treatment of intracranial hypertension in patients with acute liver failure. PMID- 9397982 TI - Quantitative liver function tests as surrogate markers for end-points in controlled clinical trials: a retrospective feasibility study. AB - Quantitative liver function tests such as the determination of galactose elimination capacity (GEC) or the aminopyrine breath test (ABT) may have the potential to serve as refined entry criteria and surrogate markers for end-points in controlled clinical trials. The magnitude of a statistically detectable difference in test results and the period of observation required to document such a difference must be known to properly design such trials. Therefore, we explored retrospectively the time course of changes in GEC and ABT and their reproducibility from a cohort of patients with alcoholic cirrhosis followed for 12 to 42 months, with a median of 34 months. In 15 patients who stopped drinking, GEC improved significantly by 0.64 mg/min/kg within 1 year (mean; 95% confidence interval [CI]: 0.42; 0.86). In contrast, it deteriorated by 0.53 mg/min/kg within 1 year (95% CI: 0.32; 0.74) in another 17 patients who continued to drink (P < .01). The residual standard deviation of the changes in GEC with respect to the patients' initial values was 0.43 mg/min/kg (95% CI: 0.32; 0.52). In addition, ABT improved significantly by 0.14% dose x kg/mmol CO2 (95% CI: 0.09; 0.18) in the abstinent group, and deteriorated by 0.09% dose x kg/mmol CO2 (95% CI: 0.06; 0.13) in the nonabstinent group (P < .01). The residual standard deviation in the above sense for ABT was 0.08% dose x kg/mmol CO2 (95% CI: 0.06; 0.10). These data indicate that clinical trials with a sample size of n = 20 in each group must achieve absolute differences (ADs) in GEC of 0.6 mg/min/kg and of 0.7 mg/min/kg to reach statistical significance at the 5% and 1% level, respectively. In the present study, a period of 11 and 12 months was necessary to observe such differences. The corresponding results for the ABT are 0.11% dose x kg/mmol CO2 (9 months of follow-up; 5% level) and 0.13% dose x kg/mmol CO2 (11 months of observation; 1% level), respectively. Provided that patients with liver diseases treated with drugs are similar to the abstinent and nonabstinent patients with alcoholic liver disease investigated in this study, such numbers could serve for the planning of controlled clinical trials, in which the control group is likely to deteriorate and the treated group is expected to improve. Trials based on GEC or ABT would require only 37 or 30 patient years of observation compared with a median of 444 patient years (range, 50-2,100 patient years) reported for various published controlled clinical trials using survival analysis. PMID- 9397983 TI - Coexpression of C-myc and transforming growth factor alfa in the liver promotes early replicative senescence and diminishes regenerative capacity after partial hepatectomy in transgenic mice. AB - We have recently shown that overexpression of c-myc and transforming growth factor alpha (TGF-alpha) in the liver of double-transgenic mice results in severe DNA damage, aberrant hepatic growth, and development of tumors at a much younger age than that observed in c-myc single-transgenic mice. We now report that double transgenic TGF-alpha/c-myc hepatocytes rapidly lose their ability to proliferate upon mitogenic stimulation following partial hepatectomy (PH). At 4 weeks of age, the overall rate of bromodeoxyuridine (BrdU) incorporation following PH was comparable in c-myc and TGF-alpha/c-myc livers and exceeded that seen in wild type (WT) mice. However, by 10 weeks of age, c-myc single-transgenic hepatocytes showed proliferative advantages over the WT cells, whereas TGF-alpha/c-myc double transgenic hepatocytes had a decreased capacity to proliferate upon mitogenic stimulation. This decreased proliferative response was accompanied by a reduction in the total fraction of proliferating hepatocytes, as well as by a decline in the induction of cyclin A, cyclin B, and cdc2 gene expression. These data show that constitutive coexpression of c-myc and TGF-alpha accelerates age-related loss in the regenerative potential following PH, and suggest that early replicative senescence of differentiated hepatocytes may have a role in providing a selective growth advantage to initiated cell populations in this model. PMID- 9397984 TI - Exposure of primary rat hepatocytes in long-term DMSO culture to selected transition metals induces hepatocyte proliferation and formation of duct-like structures. AB - We previously showed that primary rat hepatocytes plated on a rat-tail collagen coated dish and fed a chemically-defined medium supplemented with 2% dimethylsulfoxide (DMSO) can be maintained in a well-differentiated, non replicating state for periods of several months. In this study, we show that the addition of copper, iron, and zinc to the DMSO-containing chemically defined medium induced DNA synthesis and cell replication during the first two months in culture without loss of hepatic differentiation. DNA synthesis occurred throughout the hepatocyte population without regard to cellular size. No changes were observed in properties indicative of well-differentiated hepatocytes, including cellular morphology, ultrastructure, albumin, or cytokeratin-8 expression. During the third month in culture, after the hepatocytes had become confluent, pseudoduct structures became apparent. Examination of cells lining the ducts by immunohistochemistry showed that these cells lost the ability to express albumin and stained more intensely for cytokeratin 19 and laminin. The ultrastructure of the cells lining the ducts was altered and became more characteristic of bile duct cells. Immunoelectron microscopy revealed that connexin 43, a marker of bile-duct proliferation, was expressed in the duct-like cells. We conclude that under these specific nutritive conditions, primary rat hepatocytes proliferate and, with time, begin to form duct-like structures with altered gene expression and ultrastructural properties. PMID- 9397985 TI - Human hepatic myofibroblasts increase invasiveness of hepatocellular carcinoma cells: evidence for a role of hepatocyte growth factor. AB - The stroma of hepatocellular carcinomas (HCC) is infiltrated with myofibroblasts (MFs). Preliminary in vivo data have suggested that liver MF express hepatocyte growth factor (HGF), a cytokine that has been implicated in several tumor models. Our aim was to investigate the role of MF and HGF in HCC. Cultured liver MF expressed HGF messenger RNA (mRNA) and secreted HGF in their medium, as shown by Western blot, immunoprecipitation, and enzyme-linked immunosorbent assay (ELISA). Addition of MF-conditioned medium to the HepG2 HCC cell line induced cell scattering. This was associated with a decrease in cell proliferation. MF also increased about 100-fold the ability of HepG2 to invade Matrigel. Increased invasiveness was also shown for HuH7 cells, but no scattering was observed and cell proliferation was stimulated. All the effects of MF on both tumor cell types were blocked by addition of an antibody to HGF and they all could be reproduced by adding recombinant HGF to the tumor cells. RT-PCR and Western blot analysis confirmed that both tumor cell lines expressed c-met, the receptor for HGF. The effects of MF-conditioned medium were not reproduced by acidic fibroblast growth factor, basic fibroblast growth factor, epidermal growth factor (EGF), transforming growth factor-beta1 (TGF-beta1), or platelet-derived growth factor (PDGF-BB). Reverse transcription-polymerase chain reaction (RT-PCR) analysis confirmed that HGF was expressed in human HCC. Our data show that human liver MF act on HCC cells to increase their invasiveness and suggest that MF-derived HGF could be involved in the pathogenesis of HCC. PMID- 9397986 TI - Modulation of liver canalicular transport processes by the tyrosine-kinase inhibitor genistein: implications of genistein metabolism in the rat. AB - Rat liver cells express the multispecific organic anion transporter (cmoat, cmrp, mrp2) and P-glycoprotein (Pgp) in their canalicular membranes, proteins that are homologous to the multidrug-resistance related protein (MRP) and multidrug resistance (MDR) gene products in multidrug resistant tumor cells. We tested whether genistein, a modulator of drug resistance in tumor cells, affects biliary secretion of substrates of canalicular multispecific organic anion transporter (cmoat) (glucuronides of bilirubin and rhodamine, glutathione conjugate of bromsulphthalein) and of P-glycoprotein (Pgp) (rhodamine), respectively. Using the isolated perfused rat liver of control Wistar rats (TR+) and of a mutant strain (TR-) that expresses Pgp but not cmoat, we show that genistein effectively inhibits the secretion of anionic substrates of cmoat in Wistar rats but stimulates secretion of cationic rhodamine in TR- rats. Genistein is subject to glucuronidation and sulfatation and secretion of genistein and its metabolites stimulates bile flow in Wistar rats, but secretion is nearly absent in TR- rats. Because genistein and its metabolites are substrates for cmoat, inhibition of anion secretion by genistein is partially explained by competition for this transporter. Genistein is also a substrate of uridindiphosphate (UDP) glucuronyltransferase isoenzyme(s). Inhibition of glucuronidation reduces the availability of bilirubin and rhodamine glucuronates for transport via cmoat, but unconjugated cationic rhodamine becomes available for transport via Pgp at an increased cellular concentration. Daidzein, a genistein analogue with no effect on protein tyrosine kinase (PTK) shows Similar effects on secretion of organic anions and cations supporting the conclusion that genistein affects transport in liver mainly through competition with other substrates at the sites of glucuronidation and transport via cmoat. PMID- 9397987 TI - Presence of distinct AP-1 dimers in normal and transformed rat hepatocytes under basal conditions and after epidermal growth factor stimulation. AB - Activation of the transcriptional regulator AP-1, a dimeric complex formed of various combinations of Fos and Jun proteins, is a key step in the cellular response to mitogens. Because different dimers are believed to display different regulatory functions, we hypothesized that transformed cells that lack normal growth constraints might display AP-1 dimers that are different from those of normal cells. We therefore compared in primary and transformed rat hepatocytes (1) the composition of AP-1 dimers under basal conditions and (2) AP-1 induction by epidermal growth factor (EGF). Under basal conditions, AP-1 contained predominantly Jun homodimers in both cell types. However, whereas normal hepatocytes contained only JunD, both JunD and JunB were present in the AP-1 complex of 7777 cells. EGF treatment triggered almost identical programs of fos and jun gene activation at the messenger RNA (mRNA) level in both cell types, with an early accumulation of c-fos, c-jun, and junB mRNAs, but no change in junD mRNA levels. In both cell types, c-Fos and Fra-1 proteins increased after EGF treatment, but differences in the induction of Jun proteins were noted, with an increase of c-Jun in hepatocytes and an increase of JunB in 7777 cells. In both cell types, activation of AP-1 DNA binding activity by EGF was accompanied by the recruitment of Fra-1 into AP-1, detected earlier in 7777 cells than in hepatocytes, and with the transient appearance of c-Fos in 7777 cells only. Finally, EGF activated AP-1-dependent transcription in 7777 cells but not in hepatocytes. These data indicate important differences in the functional activity of AP-1 in transformed hepatocytes. PMID- 9397988 TI - Altered expression of mitogen-activated protein kinases in a rat model of experimental hepatocellular carcinoma. AB - The mitogen-activated protein kinase (MAPK) cascade acts as a focal point for signal transduction following activation of both G-protein-linked and tyrosine kinase growth factor receptors. A common intermediate between both of these diverse receptor subtypes includes the small guanosine triphosphate (GTP)-binding protein, p21ras. Point mutations of p21ras have been identified in various tumor types and lead to constitutive activation of this protein and subsequent activation of downstream pathways including the MAPK cascade. Using an in vivo model of hepatocellular carcinoma (HCC), we investigated the abundance and function of individual components of the MAPK cascade and the presence of specific p21ras mutations in this model. Expression of components of the MAPK cascade were determined in tumor and adjacent, non-neoplastic liver specimens by Western blot analysis and functional activity confirmed by substrate phosphorylation assays. Mutations in p21ras were analyzed using an enzyme-linked immunosorbent assay. In tumor, extracellular regulated kinases (ERKs) ERK1, ERK2, and mitogen-activated ERK-regulated kinase-1 (MEK1) were elevated by three- to fourfold as compared with adjacent nontumorigenic normal liver. In contrast, MEK2 was elevated by only 28%. Substrate phosphorylation and detection of phosphorylated ERK1/2 proteins showed increased functional activity of these proteins of the same magnitude as that observed for protein expression. Mutations in p21ras were not detected in this experimental model of HCC. We conclude that HCC is associated with marked changes in expression and function of components of the MAPK cascade independent of common p21ras mutations. PMID- 9397989 TI - Chromosomal aberrations in hepatocellular carcinomas: relationship with pathological features. AB - Fluorescence in situ hybridization performed on tissue sections can reveal chromosomal abnormalities related to histopathological features. This technique was performed on serial frozen sections from seven normal livers and 29 hepatocellular carcinomas (HCCs) using pericentromeric repeat-specific probes for chromosomes 1, 4, 6, 7, 8, 16, and 17. For each HCC and each probe, the percentage of cells showing one, two, or more than two signals was counted and compared with the distribution in the normal liver. According to these results, HCCs were categorized as monosomic, disomic, or polysomic (more than two signals) for the chromosome tested. These data were compared with the main histopathological characteristics of HCC. Chromosome gains were very common, preferentially affecting chromosome 1 (23 of 27 cases, 85%), chromosome 16 (16 of 27 cases, 59%), chromosome 7 (16 of 29 cases, 55%), chromosome 6 (15 of 29 cases, 52%) and chromosome 8 (14 of 29 cases, 48%). Monosomy was seen more rarely, affecting preferentially chromosome 16 (19%), chromosome 17 (14%), and chromosome 4 (10%). A significant correlation was observed between aneusomy of chromosome 4 and tumor size (P < .05) or the presence of vascular embolism (P < .05). In conclusion, chromosomal gains are frequent genetic events in human HCC. A significant association between a gain in chromosome 4 and large tumor size or vascular embolism suggests that this genetic abnormality is a late event in liver carcinogenesis. PMID- 9397990 TI - Role of Kupffer cells in gut ischemia/reperfusion-induced hepatic microvascular dysfunction in mice. AB - Kupffer cells (KCs) have been implicated in the leukocyte recruitment and microvascular dysfunction associated with liver inflammation. The overall objective of this study was to assess the role of KCs in the leukocyte adhesion and oxidative stress elicited in the liver by gut ischemia/reperfusion (I/R). The accumulation of rhodamine-6G-labeled leukocytes and the number of nonperfused sinusoids (NPS) were monitored (by intravital microscopy) in mouse liver for 1 hour after a 15-minute period of normothermic intestinal ischemia. Autofluorescence of pyridine nucleotide [NAD(P)H] was measured as an index of mitochondrial O2 consumption and redox status. Leukostasis, as well as increases in NPS and NAD(P)H autofluorescence (indicating hypoxia), were observed in the liver at 60 minutes after gut I/R. Pretreatment with gadolinium chloride (GdCl3), which reduces KC function, attenuated the liver leukostasis and NPS elicited by gut I/R. The platelet activating factor (PAF) antagonist, WEB2086, and a tumor necrosis factor (TNF)-alpha-specific antibody were also effective in attenuating the gut I/R-induced leukostasis and NAD(P)H autofluorescence. The findings of this study suggest that KCs play an important role in mediating the leukocyte recruitment, impaired sinusoidal perfusion, and tissue hypoxia elicited in the liver after gut I/R. These KC-mediated responses appear to involve the participation of both PAF and TNF-alpha. PMID- 9397991 TI - Hepatic expression of c-Myb in chronic human liver disease. AB - C-Myb is a sequence-specific DNA binding protein that regulates the expression of genes involved in cell proliferation and differentiation. The present study was designed to elucidate the role of c-Myb in the pathogenesis of chronic liver disease by an immunohistochemical approach. In normal (control) livers, few or no hepatic cells were positive for c-Myb. In livers from patients with chronic viral hepatitis, positive staining for c-Myb was found not only in spindle-shaped mesenchymal cells in expanded portal areas, but also in perisinusoidal cells (PSCs) within liver lobules. In cirrhotic livers, a few PSCs within lobules were positive for c-Myb, while no staining was seen in fibrous septa. Immunoelectron microscopy revealed that c-Myb-positive PSCs displayed morphological features of hepatic stellate cells. Other sinusoidal lining cells including Kupffer cells and sinusoidal endothelial cells, as well as hepatocytes, were all negative for c Myb. Dual c-Myb/alpha-smooth muscle actin (alphaSMA) staining revealed that more than 97% of c-Myb-positive cells were alphaSMA-positive. Moreover, dual c Myb/proliferating cell nuclear antigen (PCNA) staining showed that approximately 70% of c-Myb-positive cells also expressed PCNA. The labeling index (LI) (number of c-Myb-positive cells/0.1 mm2) significantly correlated with serum transaminase concentrations and increased in parallel with the disease activity. However, the LI showed no correlation with the degree of fibrosis. These results suggest that c-Myb may be involved in stellate cell activation and proliferation in chronically diseased human livers, and that the level of c-Myb expression is associated with the activity of chronic hepatitis. PMID- 9397992 TI - Interleukin-6 protects liver against warm ischemia/reperfusion injury and promotes hepatocyte proliferation in the rodent. AB - Interleukin-6 (IL-6) is an acute reactant cytokine with anti-inflammatory properties, which has been found to prevent injury in a model of acute hepatitis in mice through downregulation of tumor necrosis factor alpha (TNF-alpha); to correlate inversely with markers of hepatocellular injury in patients with liver ischemia; and to initiate liver regeneration in mice. In this study, we investigated the role of IL-6 in rodent models of hepatic warm ischemia/reperfusion (WI/Rp) injury. IL-6-deficient mice (-/-) were subjected to hepatic WI and compared with C57BL/6 mice, as well as IL-6 -/- mice pretreated with recombinant IL-6 (rIL-6). The effects of rIL-6 following various periods of ischemia were further studied in models of hepatic ischemia in rats. IL-6 -/- mice had increased reperfusion injury as assessed by transaminase levels and a tissue necrosis scoring system when compared with controls, an effect prevented by pretreatment with rIL-6. Similarly, rats pretreated with rIL-6 had reduced reperfusion injury and better survival than controls in each respective WI group. Tissue TNF-alpha expression measured by Northern blot analysis and serum C reactive protein (CRP) levels, a marker of inflammation, were significantly reduced in animals pretreated with rIL-6. Administration of antibodies to TNF alpha reproduced the beneficial effect of rIL-6. Hepatocyte proliferation, as assessed by a scoring method for mitotic index and proliferating nuclear cell antigen staining, was markedly increased in rIL-6-treated rats when compared with controls. In conclusion, this study suggests that IL-6 could play an important role in limiting hepatic warm ischemia/reperfusion (WI/Rp) injury, probably through its anti-inflammatory properties, modulation of TNF-alpha, and/or promotion of liver regeneration. rIL-6 might become an important cytokine in clinical situations associated with WI/Rp injury. PMID- 9397993 TI - Dual expression of matrix metalloproteinase-2 and membrane-type 1-matrix metalloproteinase in fibrotic human livers. AB - We have previously reported increased expression of matrix metalloproteinase-2 (MMP-2) using a rat model of liver fibrosis. However we did not clarify how the precursor of MMP-2 (proMMP-2) was activated. Therefore, we used human liver specimens with chronic hepatitis (CH) and liver cirrhosis (LC) to examine expression of membrane-type-1-MMP (MT1-MMP), which has recently been determined to activate proMMP-2. Northern hybridization studies showed a 5.4- and 1.4-fold increase in MMP-2 expression in CH and LC, respectively, as compared with normal liver. MT1-MMP gene expression simultaneously increased 4.0- and 1.4-fold in CH and LC, respectively. In situ hybridization using 35S-cRNA probes of MMP-2 and MT1-MMP showed prominent silver granules in elongated cells found in the lobules, periportal areas, and fibrous septa of CH and LC samples. These elongated cells expressed alpha-smooth muscle actin by immunohistochemistry. Immunoelectron microscopic examination localized MMP-2 and MT1-MMP to the rough endoplasmic reticulum of stellate cells located in the lobules and periportal areas, or to fibroblasts in the fibrous septa, suggesting that MMP-2 and MT1-MMP were produced by these cells. In addition, cytoplasmic and membranous immunodeposits of both MMPs were found in endothelial cells, Kupffer cells, capillary endothelial cells, and lymphocytes, indicating that activation of proMMP-2 occurs locally. Increased expression of MMP-2 and MT1-MMP was detected in CH and LC, while dual over expression was found in stellate cells and fibroblasts, possibly resulting in the increase of active MMP-2 in and around these cells. These findings suggest that activated MMP-2 may remodel liver parenchyma during the process of liver fibrosis. PMID- 9397994 TI - Antibodies to tumor necrosis factor alfa attenuate hepatic necrosis and inflammation caused by chronic exposure to ethanol in the rat. AB - Tumor necrosis factor (TNF)alpha, a pivotal cytokine involved in inflammation, is produced primarily by Kupffer cells in the liver. It has been shown that inactivation of Kupffer cells prevents alcohol-induced liver injury; therefore, the purpose of this study was to determine if neutralizing anti-TNF-alpha antibody is also effective. Male Wistar rats were exposed to ethanol (11 to 12 g x kg(-1) x d[-1]) continuously for up to 4 weeks via intragastric feeding using an enteral feeding model. Before ethanol exposure, polyclonal anti-mouse TNF alpha rabbit serum was injected (2.0 mg/kg intravenously). There were no significant differences in body weight, mean ethanol concentration, or cyclic patterns of ethanol in urine when ethanol- and ethanol plus antibody-treated groups were compared. Expression of TNF-alpha and macrophage inflammatory protein 2 (MIP-2) messenger RNA (mRNA), determined using reverse transcription-polymerase chain reaction, was three- to four-fold higher in livers of ethanol-treated rats than in those of rats fed an ethanol-free, high-fat control diet. In addition, MIP-2 levels were also elevated when detected by Northern blot analysis. Anti-TNF alpha antibody did not affect expression of mRNA for interleukin (IL) 1alpha, IL 6, transforming growth factor beta1, or TNF-alpha. However, MIP-2 mRNA expression, which is regulated by TNF-alpha, was decreased significantly by anti TNF-alpha antibody treatment. Serum aspartate transaminase levels were elevated in ethanol-treated rats to 136 +/- 12 IU/L after 4 weeks but only reached 90 +/- 5 IU/L (P < .05) in rats treated with anti-TNF-alpha antibody. The hepatic inflammation and necrosis observed in ethanol-fed rats were attenuated significantly by antibody treatment, and steatosis was not. These results support the hypothesis that TNF-alpha plays an important role in inflammation and necrosis in alcohol-induced liver injury and that treatment with anti-TNF-alpha antibody may be therapeutically useful in this disease. PMID- 9397995 TI - Dietary saturated fatty acids down-regulate cyclooxygenase-2 and tumor necrosis factor alfa and reverse fibrosis in alcohol-induced liver disease in the rat. AB - We investigated the potential of dietary saturated fatty acids to decrease endotoxemia and suppress expression of cyclooxygenase 2 (Cox-2) and tumor necrosis factor alpha (TNF-alpha) in established alcohol-induced liver injury. Six groups (five rats/group) of male Wistar rats were studied. Rats in group 1 were fed a fish oil-ethanol diet for 6 weeks. Rats in groups 2, 3, and 4 were fed fish oil and ethanol for 6 weeks. Ethanol administration was stopped at this time, and the rats were switched to isocaloric diets containing dextrose with fish oil (group 2), palm oil (group 3), or medium-chain triglycerides (group 4) as the source of fat for an additional 2 weeks. Rats in groups 5 and 6 were fed fish oil-ethanol and fish oil-dextrose, respectively, for 8 weeks. Liver samples were analyzed for histopathology, lipid peroxidation, and levels of messenger RNA (mRNA) for Cox-2 and TNF-alpha. Concentrations of endotoxin were determined in plasma. The most severe inflammation and fibrosis were detected in groups 1 and 5, as were the highest levels of endotoxin, lipid peroxidation, and mRNA for Cox 2 and TNF-alpha. After ethanol was discontinued, there was minimal histological improvement in group 2 but near normalization of the histology, including regression of fibrosis, in groups 3 and 4. Histological improvement was associated with decreased levels of endotoxin, lipid peroxidation, and reduced expression of Cox-2 and TNF-alpha. The data indicate that a diet enriched in saturated fatty acids (groups 3 and 4) effectively reverses alcohol-induced liver injury, including fibrosis. The therapeutic effects of saturated fatty acids may be explained, at least in part, by reduced endotoxemia and lipid peroxidation, which in turn result in decreased levels of TNF-alpha and Cox-2. PMID- 9397996 TI - Enhanced expression of cytokine-induced neutrophil chemoattractant in rat hepatic allografts during acute rejection. AB - The kinetics of messenger RNA (mRNA) and protein levels of cytokine-induced neutrophil chemoattractant (CINC) in rat hepatic allografts during acute rejection were investigated. Infiltrating cells were identified by double immunostaining with anti-CINC and anti-macrophage monoclonal antibodies, ED1 and ED2. The serum CINC concentration in untreated hepatic allograft recipients increased significantly at a constant rate over time after transplantation. No significant increases in serum CINC concentrations were observed in hepatic isografts or allografts treated with the immunosuppressant FK506. The number of neutrophils in untreated hepatic allografts increased significantly at a constant rate. Conversely, neutrophil accumulation in isografts or allografts treated with FK506 was much less than in untreated hepaticallografts. Immunostaining revealed that in the portal areas, mononuclear cells infiltrating untreated allograft liver were mainly positive for CINC and that CINC+ cells represented a subpopulation (approximately 25%) of the ED1+ cells. On the other hand, in the sinusoidal areas CINC+ cells were scattered and mainly positive for ED2. Levels of CINC mRNA in liver tissues taken from untreated hepatic allografts increased after transplantation, peaked on day 5, and decreased thereafter. Hepatic allografts treated with FK506 or isografts showed much lower levels of CINC mRNA than untreated allografts. Allogeneic mixed lymphocyte reactions induced CINC production. The cellular source of CINC was mononuclear cells. CINC production in mixed lymphocyte reactions was inhibited in the presence of anti-tumor necrosis factor alpha (TNF-alpha) antibody. These results suggest that enhanced expression of CINC mRNA and prominent accumulation of neutrophils in the liver grafts are characteristic features of the immune response during acute rejection. PMID- 9397997 TI - Targeting naproxen coupled to human serum albumin to nonparenchymal cells reduces endotoxin-induced mortality in rats with biliary cirrhosis. AB - Endotoxin is thought to play a major role in cirrhotic liver disease. Cyclo oxygenase inhibitors were shown to be partially protective against endotoxin but cannot be used in cirrhotic patients because of renal side-effects. We argued that administration of naproxen (NAP) linked to human serum albumin (HSA), which results in specific delivery of NAP to endothelial cells (EC) and Kupffer cells (KC) and exhibited hepatoprotective effects against lipopolysaccharide (LPS) in vitro, could protect cirrhotic rats from LPS toxicity while preserving renal function. The studies were performed in rats rendered cirrhotic by bile duct ligation (BDL); animals received LPS (Escherichia coli, 800 microg/kg) intravenously. Five groups were studied: LPS alone, rats pretreated with a conventional dose of NAP (50 mg/kg), NAP-HSA (22 mg/kg), NAP equimolar to NAP-HSA (1.5 mg/kg), or the HSA carrier. LPS induced significant mortality (55%); this was not affected by equimolar NAP (57%) but accentuated by conventional NAP (88%). In contrast, NAP-HSA provided significant protection (9%; P < .05). After conventional NAP treatment, significant renal toxicity was observed as evidenced by a marked reduction in sodium excretion (LPS vs. NAP-HSA vs. NAP [50 mg/kg] 33 +/- 22 vs. 50 +/- 39 vs. 4 +/- 3 micromol/h; P < .05). Renal prostaglandin E2 (PGE2) excretion was reduced by NAP in all groups, but most markedly at the conventional dosage (LPS vs. NAP-HSA vs. NAP [50 mg/kg] 132 +/- 115 vs. 39 +/- 19 vs. 9 +/- 8 ng/mL; P < .05). Successful targeting was evidenced by a significant hepatic enrichment of NAP in the NAP-HSA group compared with the equimolar untargeted group (30.16 +/- 9.33 vs. 1.13 +/- 1.95 nmol/g liver). Thus, targeting NAP to EC/KC results in improved survival, higher efficacy, and sparing of renal function in cirrhotic rats. PMID- 9397998 TI - Cytoprotection by the osmolytes betaine and taurine in ischemia-reoxygenation injury in the perfused rat liver. AB - Medium osmolarity sensitively regulates Kupffer cell functions like phagocytosis and prostaglandin (PG) and cytokine production. Betaine and taurine, recently identified as osmolytes in liver cells, interfere with these effects. Because Kupffer cell activation is an important pathogenic mechanism in ischemia reoxygenation injury, the influence of osmolarity and osmolytes was investigated in a rat liver perfusion model of warm ischemia. Livers were perfused with different medium osmolarities for 60 to 90 minutes in the absence of oxygen, followed by another 90 minutes of reoxygenation. Lactate dehydrogenase (LDH) leakage into the effluent perfusate during the hypoxic and the reoxygenation period was eight- to 10-fold higher with a medium osmolarity of 385 mosmol/L than in normo-osmolarity, and further decreased with hypo-osmolar perfusion buffer. Betaine and taurine addition to the perfusate in near physiological concentrations decreased hypoxia-reoxygenation-induced LDH leakage, aspartate transaminase (AST) leakage, and perfusion pressure increase in hyperosmolar and normo-osmolar perfusions. Stimulation of PGD2, PGE2, thromboxane B2 (TXB2), and tumor necrosis factor alpha (TNF-alpha) release, as well as induction of carbon uptake by the liver during reoxygenation, were suppressed by betaine and taurine, pointing to an interference of these osmolytes with Kupffer cell function. In contrast, endothelial cell function as assessed by hyaluronic acid (HA) uptake was not influenced. It is concluded that warm ischemia-reoxygenation injury in rat liver is aggravated by hyperosmolarity and attenuated by hypo-osmolarity. The osmolytes betaine and taurine have a protective effect, presumably by inhibition of Kupffer cell activation. PMID- 9397999 TI - Neonatal granulocytosis is a postpartum event which is seen in the liver as well as in the blood. AB - In a recent series of studies, we demonstrated that stress in humans and animals, with resultant sympathetic nerve strain, induces severe granulocytosis, because granulocytes carry adrenergic receptors on the surface. Because activated granulocytes produce free radicals and superoxides, they sometimes induce tissue damage if the stress is too strong or continuous. Human neonates are also known to show high levels of granulocytes in the peripheral blood. In this study, we investigated whether such neonatal granulocytosis are a stress-associated response at birth. Both human and mouse materials, before and after birth, were used. The number of leukocytes in the blood, as well as some other factors in the serum, were measured. Although levels of granulocytes were found to be low in fetal humans and mice, they increased sharply after birth. In parallel with this postpartal granulocytosis, transaminases in sera increased transiently. In reference to results of a transient elevation in the levels of catecholamines at birth in mice, all these phenomena resemble stress-associated responses. Indeed, fatty liver and hematopoietic destruction in the liver were also observed in mice and humans. At this time, the production of inducible nitric oxide synthase (iNOS) by granulocytes in the liver was evident. These results suggest that neonatal granulocytosis is a postpartum event which results from various stresses (e.g., oxygen stress) at birth. This event may be responsible for such well-known neonatal phenomena as the termination of fetal hematopoiesis in the liver and as neonatal jaundice. PMID- 9398000 TI - Progesterone metabolites and bile acids in serum of patients with intrahepatic cholestasis of pregnancy: effect of ursodeoxycholic acid therapy. AB - The concentrations in serum of sulfated metabolites of progesterone are known to be elevated in patients with intrahepatic cholestasis of pregnancy (ICP). The profiles of these metabolites and conjugated bile acids were analyzed in serum from 11 patients with ICP before and during administration of ursodeoxycholic acid (UDCA) (8 patients) or placebo (3 patients). The clinical condition of 7 of the patients given UDCA improved markedly, and 1 patient given placebo had a spontaneous remission of the disease. The total concentration of conjugated bile acids in the 11 patients was 25 +/- 6 micromol/L (mean +/- SEM) and decreased to 6.3 +/- 3.5 micromol/L in the 7 patients responding to treatment with UDCA. The level of 7alpha-hydroxy-4-cholesten-3-one was significantly lower (7.2 +/- 2.2 ng/mL) in patients with ICP than in healthy pregnancy (18 +/- 4.6 ng/mL) (P < .05). The concentrations of 5alpha-pregnane-3alpha,20alpha-diol mono- and disulfates decreased by 52% +/- 7.9% and 68% +/- 5.5%, respectively, in the patients responding to treatment. Similar decreases were observed for the mono- and disulfates of 5alpha-pregnane-3alpha,20alpha,21-triol and 5beta-pregnane 3alpha,20alpha-diol. The disulfate of 5alpha-pregnane-3beta,20alpha-diol showed a smaller decrease, while glucuronidated steroids were not affected. The 3alpha /3beta-hydroxysteroid ratio and di-/monosulfate ratio decreased significantly during UDCA. The magnitudes of the changes of bile acid and steroid concentrations during UDCA were not correlated to each other. The results suggest that UDCA stimulates the biliary excretion of steroids with a 3alpha-sulfoxy group and disulfates. This effect seems to be independent of the effect on bile acid excretion, indicating the use of different transport proteins. The possibility of an effect of UDCA on the formation of the steroid sulfates cannot be excluded. PMID- 9398001 TI - Modulation of circadian expression of D-site binding protein by the schedule of parenteral nutrition in rat liver. AB - The aim of this study was to investigate the changes in the circadian rhythm of the expression of liver-specific genes caused by different schedules of parenteral nutrition (PN). Rats received PN continuously throughout the day or intermittently during the night or day for 7 days. They were examined for gene expression of D-site binding protein (DBP), albumin, and cytochrome P450 cholesterol 7alpha-hydroxylase (CYP7) in the liver. The nocturnal PN group showed circadian expression of DBP messenger RNA (mRNA) and protein with a peak at 10 PM, in the same manner as the control rats receiving normal chow feeding. However, the diurnal PN group showed inverted expression of DBP mRNA and protein with a peak at 10 AM. CYP7 mRNA levels exhibited good synchronization with the levels of DBP mRNA in all groups, whereas albumin mRNA levels did not show such synchronization. Gel mobility-shift assay disclosed that the binding activity of the nuclear extracts to the CYP7 gene promoter was changed by the PN schedule in accordance with the expression of CYP7 mRNA. The PN schedule modulates the circadian rhythm of DBP expression and may have an effect on hepatic bile acid formation through transcriptional regulation of the CYP7 gene. PMID- 9398002 TI - Effect of cytochrome P450 induction on phosphorus metabolites and proton relaxation times measured by in vivo 31P-magnetic resonance spectroscopy and 1H magnetic resonance relaxometry in human liver. AB - Experimental and clinical studies have led to the hypothesis that the phosphodiester signal obtained by 31P magnetic resonance (MR) spectroscopy may be a specific marker for the hepatic induction of oxidative metabolism (P450 induction) by phenobarbitone or ethanol. Systematic studies in humans are lacking. Therefore, we studied 10 volunteers who received rifampin (600 mg/d) for 6 days, resulting in a documented induction of oxidative metabolism as measured by an increase in urinary 6-beta-hydroxycortisol output in all volunteers (P = .0004). 31P-MR spectroscopy and 1H-MR relaxometry were performed before and after hepatic P450 induction. As shown by 31P-MR spectroscopy, the median phosphomonoester concentration (PME) relative to nucleoside triphosphate (NTP) increased by 21% from 0.63 (range, 0.40-0.89) before induction to 0.76 (0.49 1.67) after induction (P = .0451). The median level of phosphodiesters (PDE) relative to NTP increased by 28% from 4.82 (3.41-6.67) before induction to 6.18 (4.63-11.63) after induction (P = .0091). An increase in the level of inorganic phosphates (Pi) relative to NTP was observed, but changes were not significant. As shown by 1H-MR relaxometry, a nonsignificant trend of the liver parenchyma to shorter relaxation times was observed after P-450 induction. In conclusion, both PME/NTP and PDE/NTP ratios (measured by in vivo 31P-MR spectroscopy) increased significantly after hepatic induction with rifampin. Further clinical studies with 31P-MR spectroscopy must take into account the potential effects of P450 inducing agents. PMID- 9398003 TI - An improved digitonin-collagenase perfusion technique for the isolation of periportal and perivenous hepatocytes from a single rat liver: physiological implications for lobular heterogeneity. AB - Morphological and functional heterogeneity of hepatocytes according to their position in the liver lobule has been known for many years. The digitonin collagenase perfusion technique is widely used to study hepatocyte heterogeneity and has yielded reliable data. However, with this procedure, periportal (PP) or perivenous (PV) hepatocytes are isolated from different livers, allowing only comparison between cell populations issued from two separate animals. To overcome this drawback, we have modified this technique by perfusing the two main rat liver lobes of a single animal in succession. The procedure involved alternate clamping of the median and the left lateral lobes, restricting digitonin infusion to one lobe via the portal vein, and to the other via the caudal vena cava. Lobe exclusion during digitonin perfusion, and zonal restriction of digitonin-induced damage, were monitored using macroscopic and histological controls. We compared our results with previous data on PP and PV hepatocytes issued from two different livers using the conventional digitonin-collagenase perfusion technique. First, we found that the cellular sensitivity to angiotensin II, a calcium-mobilizing agonist, was 60% to 80% higher in PV than in PP hepatocytes, whereas, previously, no difference had been recorded. Second, we found that albumin messenger RNAs (mRNAs) were 35% more abundant in PP than in PV hepatocytes, whereas, previously, larger differences had been reported. Our results show that PP and PV hepatocytes may be isolated from a single liver using an improved digitonin-collagenase perfusion technique. Furthermore, we suggest that zonal differences can be artificially masked or amplified when comparing PP and PV cell populations from two different livers, indicating that it is preferable to use a single liver for accurate zonal comparisons between hepatocytes. PMID- 9398004 TI - Hepatic growth hormone receptor, insulin-like growth factor I, and insulin-like growth factor-binding protein messenger RNA expression in pediatric liver disease. AB - Major changes in serum levels of insulin-like growth factor I (IGF-I) and IGF binding proteins (IGFBPs) occur in children with end-stage liver disease in association with changes in body composition. We hypothesized that these changes would be associated with changes in hepatic messenger RNA (mRNA) expression. Eleven children with end-stage extrahepatic biliary atresia and 11 controls (liver donors) were studied. Serum samples were obtained from the children with biliary atresia immediately before orthotopic liver transplantation. Serum IGF-I, IGFBP-1, and IGFBP-2 levels were measured by radioimmunoassay, and IGFBP-3 by immunoradiometric assay. In both groups, growth hormone receptor mRNA expression was examined by quantitative reverse transcription-polymerase chain reaction, IGF I mRNA expression by ribonuclease protection assay, and IGFBP-1 to -4 mRNA expression by Northern analysis. Growth hormone receptor and IGF-I mRNA levels were reduced 1.7-fold (P = .003) and 9.6-fold (P = .0001) in biliary atresia compared with levels in controls. Despite increased serum IGFBP-1 levels and reduced IGFBP-3 levels in biliary atresia, there was no change in either IGFBP-1 or IGFBP-3 mRNA expression. In contrast, serum levels and mRNA expression of IGFBP-2 were increased 1.6-fold (P = .003) and twofold (P = .0001), respectively, compared with controls. Gene expression did not correlate with liver dysfunction or body composition. Changes in growth hormone receptor and IGF-I mRNA expression may account for the reduction in serum IGF-I found in pediatric liver disease. In contrast, the marked alteration in circulating IGFBP levels was not accompanied by changes in hepatic IGFBP gene expression, suggesting that posttranslational mechanisms may be important. PMID- 9398005 TI - Hepatic expression of the woodchuck hepatitis virus X-antigen during acute and chronic infection and detection of a woodchuck hepatitis virus X-antigen antibody response. AB - The expression and localization of the woodchuck hepatitis virus X-antigen (WHxAg) was examined and compared with other markers of a woodchuck hepatitis virus (WHV) infection using rabbit antisera generated against recombinant WHxAg produced in bacteria. Cellular fractionation studies showed that WHxAg was localized to the soluble and cytoskeletal fractions of the cell when assayed by immunoprecipitation of [35S]-met-cys labeled extracts derived from primary cultures of acute WHV-infected hepatocytes. Immunohistochemical examination of liver from chronic WHV-infected animals showed WHV core antigen (WHcAg) and WHxAg expression in non-neoplastic tissue. The WHxAg was found localized to the cytoplasm of infected cells, similar to WHcAg. WHxAg expression was diminished in the foci of altered hepatocytes and in hepatocellular adenomas but was found in only 1 of 11 hepatocellular carcinomas (HCC). Hepatic biopsies from woodchucks experimentally inoculated with WHV were examined during the acute phase of infection and during convalescence for WHcAg and WHxAg expression by immunohistochemistry. Concurrent expression of WHcAg and WHxAg was observed during the viremic phase of infection. The two antigens exhibited similar localization to the cell cytoplasm, similar distribution within the liver lobule, and similar patterns of clearance during convalescence. An immune response to WHxAg was documented in some woodchucks following acute WHV infection. These studies further define the woodchuck model of HBV infection and should allow for the investigation of the role of hepadnaviral X-antigen expression in the pathogenesis of chronic hepatitis and HCC. PMID- 9398006 TI - Nucleotide sequence variations in the internal ribosome entry site of hepatitis C virus-1b: no association with efficacy of interferon therapy or serum HCV-RNA levels. AB - The extreme 5'-proximal sequences of the hepatitis C virus (HCV) genome including the 5'untranslated region (5'UTR) and the first 30 nucleotides of the core region are highly conserved, and serve as an internal ribosome entry site (IRES) that initiates the cap-independent translation of HCV polyprotein. Mutations in the IRES sequence have been shown to cause changes in the efficiency of protein translation in vitro. However, the significance of genetic variations in the IRES is not fully known in clinical settings. Pretreatment sera of 25 patients with HCV-1b infection who were treated with interferon were amplified by polymerase chain reaction (PCR), and the IRES sequence was directly sequenced. Correlation of interferon responses or other clinical features with IRES sequence variability was studied. Eleven of 25 patients were sustained responders (SR) of interferon treatment (negative serum HCV RNA and normal alanine transaminase levels for 6 months after the end of interferon treatment), and the other 14 patients were nonresponders ([NR], defined as any patient with positive serum HCV RNA within 6 months after the end of interferon therapy). In each patient, one to four nucleotide substitutions were found compared with the consensus sequence of HCV 1b genotype. There were no differences in the number of nucleotide substitutions between either SR and NR (mean, 1.8 in SR, 2.1 in NR; P = .30), and no specific variations associated with SR or NR were observed. Although NR had significantly higher serum levels of pretreatment HCV RNA than SR (median, 16 vs. <0.5 Meq/mL; P = .02), there was no correlation between the HCV-RNA level and the number of nucleotide substitutions in the IRES (mean, 1.9 nucleotide substitutions in 12 patients with HCV RNA <0.5 Meq/ mL vs. 2.1 nucleotide substitutions in 13 patients with HCV RNA >0.5 Meq/mL; P = .61). Sequence variability of the IRES has no influence on interferon efficacy or serum HCV-RNA concentrations in patients with chronic HCV-1b infection. PMID- 9398007 TI - A randomized, controlled trial of a 24-month course of interferon alfa 2b in patients with chronic hepatitis B who had hepatitis B virus DNA without hepatitis B e antigen in serum. AB - Short-term interferon treatment of serum hepatitis B e antigen (HBeAg)-negative carriers with serum hepatitis B virus (HBV) DNA and histological features of chronic hepatitis B has been largely unsuccessful. In a pilot study of long-term treatment, 42 such patients were randomly assigned to 6 million units of interferon alfa 2b (IFN-alpha2b) three times per week for 24 consecutive months (n = 21, 4 with cirrhosis) or to no therapy (n = 21, 3 with cirrhosis). Five patients (24%) discontinued therapy because of treatment-related adverse reactions. Serum levels of alanine transaminase (ALT) became persistently normal and HBV DNA undetectable by dot-blot assay in 8 patients receiving interferon and in 2 untreated controls (38% vs. 10%; P = .03). Hepatitis flare-ups disappeared in 17 patients during therapy compared with 6 controls (81% vs. 29%; P < .001). During a median period of 22 months after interferon was stopped, 2 treated patients (10%) lost serum hepatitis B surface antigen (HBsAg) and seroconverted to antibodies to hepatitis B surface antigen (anti-HBs). Serum ALT remained persistently normal and HBV DNA undetectable by dot-blot assay in 6 initial responders and 1 initial nonresponder, compared with none of the 21 untreated controls (sustained response: 33% vs. 0; P < .001). Comparative analysis of pre- and posttreatment liver biopsies showed that mean Knodell scores dropped in the treated group (10.3 to 5.3; P = .01), but not in the untreated group (9.3 to 9.8; not significant). In conclusion, a 24-month course of treatment with 6 MU IFN alpha2b was well tolerated by most patients, led to sustained suppression of HBV in one third, and attenuated hepatitis in 81% of patients. PMID- 9398008 TI - Humoral immune response to the E2 protein of hepatitis G virus is associated with long-term recovery from infection and reveals a high frequency of hepatitis G virus exposure among healthy blood donors. AB - The second envelope protein (E2) of the hepatitis G virus (HGV) was expressed in Chinese hamster ovary (CHO) cells and showed a molecular weight of approximately 60 to 70 kd, with 15 to 25 kd of the size contributed by N-linked glycosylation. An enzyme-linked immunosorbent assay (ELISA) using HGV-E2 was developed to test for antibodies to this protein (anti-E2) in human sera. High sensitivity was achieved by developing monoclonal antibodies (mAbs) to HGV-E2, which were used as capture antibodies in the ELISA. Our studies revealed that 16% of healthy Spanish blood donors were exposed to HGV, indicating that additional routes of viral transmission besides parenteral exposure might exist. An even higher prevalence of exposure to HGV (52%-73%) was found in several groups at risk of parenteral exposure to infectious agents, i.e., intravenous drug users, transfusion history, hemophiliacs, and hepatitis C virus (HCV)-positive patients. Most anti-E2 positive patients were HGV-RNA-negative and vice versa, indicating an inverse correlation of these two viral markers. A panel of 16 posttransfusion patients followed for up to 16 years revealed that patients who develop an anti-E2 response become HGV-RNA-negative, while patients who do not develop anti-E2 are persistently infected. Immunity to HGV seems to be long-lasting, because circulating antibody to E2 could still be detected 14 years after seroconversion. Sequence comparisons showed that E2 is highly conserved among isolates collected worldwide, indicating that immune escape variants are not common in HGV infections. This reflects on a molecular level why HGV infections usually are cleared spontaneously by the host. However, possible mechanisms of HGV persistence, as found in some patients, remain to be elucidated. PMID- 9398009 TI - Frequent detection of hepatitis A viral RNA in serum during the early convalescent phase of acute hepatitis A. AB - The diagnosis of type A hepatitis is performed mainly by immunoglobulin M (IgM) anti-hepatitis A antibody assay, but it has not been established whether there is a correlation between changes in viremia and the clinical course of type A hepatitis. We examined hepatitis A virus (HAV) RNA in the sera from type A and non-A, non-B, non-C acute hepatitis and analyzed the relation of HAV viremia with alanine aminotransferase (ALT) and IgM-HA levels. Two hundred sera from 38 patients with type A acute hepatitis and 20 patients with non-A, non-B, non-C acute hepatitis were examined for the presence of HAV RNA. HAV RNA was detected by nested reverse transcription-polymerase chain reaction (RT-PCR) with primers located at the 5' non-translated region of HAV. HAV RNA was detected in 35 of 38 (92%) type A hepatitis patients and in 60 of 156 (38%) serum samples. In contrast, it was detected in none of 44 serum specimens from 20 non-A, non-B, non C acute hepatitis patients. In type A hepatitis, the mean ALT level in HAV RNA positive specimens was 1,481 +/- 2,042 (range, 20-10,370) IU/L and that in HAV RNA negative specimens 186 +/- 330 (range, 8-1,698). The positivity of HAV RNA was correlated with the level of transaminase at the time of sample collection. The mean duration from the onset of symptoms to disappearance of HAV RNA was 18 +/- 14 days. The mean titer of IgM-HA in HAV RNA positive cases was 5.0 +/- 1.4, in negative cases 5.7 +/- 1.1, with no statistical difference. Our results indicate that HAV RNA in serum is detectable in the majority of type A hepatitis cases in their early convalescent phase by nested RT-PCR. PMID- 9398010 TI - Prediction of response during interferon alfa 2b therapy in chronic hepatitis C patients using viral and biochemical characteristics: a comparison. AB - Patients with chronic hepatitis C (n = 103) were treated for 24 weeks with interferon alfa 2b and followed up for 24 weeks after cessation of therapy (week 48). When hepatitis C virus (HCV) RNA at week 48 was used to assess interferon response, 15 (14.6%) were virological complete responders, and all have remained HCV RNA negative for a mean of 3 years. At week 48, 3 of 15 virological complete responders had elevated alanine transaminase (ALT) values. When serum ALT level was used at week 48 to determine response to interferon, 20 (19.4%) were biochemical complete responders. However, 8 of the 20 patients with normal ALT levels were HCV RNA positive at week 48, and 7 of these individuals have had a recurrence of elevated ALT levels within 3 years after cessation of treatment. These findings indicate that measurement of HCV RNA was more accurate than ALT in determining true responses to interferon therapy. Identification of nonresponders early during the course of interferon treatment showed that an elevated ALT level at week 12 was 92% predictive (odds ratio 3.7) but misidentified 33% (5 of 15) of the patients who were virological complete responders at week 48. In contrast, a positive HCV RNA at week 12 of treatment was 98% predictive (odds ratio 35.5) and misidentified only 6.7% (1 of 15) of the virological complete responders. Thus, positive HCV RNA at week 12 of therapy was more accurate in identifying eventual virological nonresponders than measurement of ALT at this time. Termination of interferon therapy in patients who were HCV RNA positive at week 12 would result in a 27% reduction in the direct medical costs and keep patients from undergoing unnecessary treatment. Therefore, testing for HCV RNA at week 12 to identify nonresponders and then discontinuing their treatment is practical, cost-efficient and beneficial both to patients and to third-party payers. PMID- 9398011 TI - Histological and clinical outcome after liver transplantation for hepatitis C. AB - Hepatitis frequently recurs after liver transplantation for hepatitis C. However, the histological progression of disease, predictors of recurrence and disease severity, and patient survival remain uncertain. Fifty-five patients with cirrhosis caused by chronic hepatitis C underwent liver transplantation between January 1990 and December 1993. Hepatitis C genotype was determined, and liver biopsies were performed at frequent intervals posttransplantation. The median follow-up time was 40.4 months. The cumulative rate of survival was no different in liver transplant recipients for hepatitis C than in liver transplant recipients for other chronic liver diseases (P = .62). Histological recurrent hepatitis C developed in 33 of 50 patients assessable for disease recurrence; the median recurrence-free survival time was 13.4 months. Histological activity and stage were mild in most cases. Only 2 patients developed cirrhosis, and no patient required a second transplantation for recurrent disease. Patients with acute cellular rejection had a shorter recurrence-free survival (P = .0141). In patients with recurrent hepatitis, rejection also was correlated with increased histological grade 2 years after transplantation (P = .0061). Recurrence-free survival was decreased in patients infected with genotype 1 (1a and 1b combined) compared with genotypes 2 and 3 combined (P = .02), whereas there was no difference between genotypes 1a and 1b (P > .80). Only patients infected with genotype 1a or 1b developed bridging fibrosis or cirrhosis. In addition, patients who had an early recurrence had a greater risk of progressing to bridging fibrosis or cirrhosis (hazard ratio, 5.1; P = .0473). In our experience, recurrent hepatitiS C after liver transplantation in most cases is mild and survival is unaffected. Both acute cellular rejection and infection with genotype 1 are independent risk factors for reduced recurrence-free survival, and early recurrence is associated with a higher risk of disease progression. PMID- 9398012 TI - A case-control study on GB virus C/hepatitis G virus infection and hepatocellular carcinoma. Brescia HCC Study. AB - A new hepatitis-associated RNA virus of the Flaviviridae family has been identified and named GB virus C/ hepatitis G virus (HGV). We carried out a case control study to evaluate the association of HGV infection with hepatocellular carcinoma (HCC). We recruited 170 patients hospitalized for HCC (143 male and 27 female, mean age 64 years) and 306 patients hospitalized for nonliver diseases (controls) in Brescia, Italy. HGV RNA was detected by reverse transcription polymerase chain reaction (RT-PCR) and antibodies against HGV E2 protein (anti E2) by an immunoassay test. HGV RNA was found in 8 cases (4.7%) and 4 controls (1.3%). The relative risk (RR) for HGV RNA positivity adjusted for demographic variables and hepatitis B surface antigen (HBsAg), hepatitis C virus (HCV) RNA, and alcohol was 7.3 (95% confidence interval, 1.7-30.6; P = .009). No HGV RNA positive subject was also positive for anti-E2. Anti-E2 prevalence did not differ significantly between cases (20%) and controls (15.3%), and no RR increase was found by this marker. Among subjects with HGV exposure (HGV RNA plus anti-E2 positive), a greater proportion of cases (40%) than controls (14%) had transfusion history. The possible role of HGV in HCC etiology seems modest because the population-attributable risk is lower (4%) than those for HBsAg (22%), HCV RNA (36%), and heavy alcohol intake (52%). This study supports the hypothesis of an association between HGV infection and HCC, although at present there are insufficient data on the causality of the association. PMID- 9398013 TI - The prevalence of surface antigen variants of hepatitis B virus in Papua New Guinea, South Africa, and Sardinia. AB - Three assays, one based on monoclonal antibodies and the others on polyclonal antibodies, were employed to detect hepatitis B surface antigen (HBsAg)-reactive samples in both vaccinated and unvaccinated populations in areas of the world where hepatitis B virus (HBV) is endemic. Any discordant sera were tested by polymerase chain reaction (PCR) to confirm current infection, and sequence data were obtained from the DNA coding for the major hydrophilic region (MHR) of HBsAg of those samples positive for PCR. In all countries studied, samples that reacted in one HBsAg assay but not another were found. In the most extreme case, about 5% of viremic sera in Papua New Guinea were nonreactive in the monoclonal HBsAg assay; 9 of the 13 PCR-positive samples had novel or once-described variants, or a variant out of its usual genotype context. In South Africa, samples with sequences of subtype ayw2 reacted poorly, particularly in the polyclonal assay. Two had novel variants. In Sardinia, antibody to hepatitis B core antigen (anti HBc) was analyzed as a marker of infection. A significant proportion of anti-HBc positive, but monoclonal HBsAg-negative, vaccinees and unvaccinated persons were found to be PCR positive, as were some individuals without any markers of hepatitis B virus infection. Five more novel variants were found in these groups. There are implications for the design of HBsAg assays, which may have to be modified according to local sequence variability. Not all discordant samples were explained by variants, indicating that assay sensitivity is fundamental to diagnostic efficacy. Overall, this study defined 16 novel variants and 2 new potential epitope clusters. PMID- 9398015 TI - Surrogates, survival, and subversion. PMID- 9398014 TI - Substrate specificity of sinusoidal bile acid and organic anion uptake systems in rat and human liver. AB - The Na+-dependent bile salt uptake systems Ntcp (rodents) and NTCP (human), and the Na+-independent organic anion transporters oatpl (rat) and OATP (human) mediate sinusoidal uptake of a variety of amphipathic organic compounds into hepatocytes. Their properties indicate that an overall hepatic clearance of albumin-bound compounds is mediated by a limited number of multispecific transporters with partially overlapping substrate specificities. PMID- 9398016 TI - Selective hepatic drug delivery: from basic concept to clinical practice. PMID- 9398017 TI - A molecular basis for jaundice in intrahepatic and extrahepatic cholestasis. PMID- 9398018 TI - Traffic jam for hemochromatosis mutant protein. PMID- 9398019 TI - Lipoproteins: are they important components of host defense? PMID- 9398020 TI - Immune complexes in hepatitis C. PMID- 9398021 TI - Aplastic anemia and viral hepatitis: a second look. PMID- 9398022 TI - Predicting the risk of tumor recurrence following transplantation for hepatocellular carcinoma. PMID- 9398023 TI - Hepatitis C: viral kinetics. PMID- 9398024 TI - Prevention of variceal bleeding with band ligation. PMID- 9398025 TI - Nitric oxide-inhibitory effect of aminoguanidine on renal function in rats. AB - Inhibition of nitric oxide (NO) synthesis by structural analogues of L-arginine reduces glomerular filtration, renal blood flow, sodium excretion, and urine output. N(G)-nitro-L-arginine methyl ester (L-NAME) inhibits constitutive and inducible isoforms of NO synthase, while aminoguanidine (AG) selectively inhibits inducible isoforms of NO synthase. We assessed the NO-inhibitory activity of AG on renal function. Rats were treated with aminoguanidine 50 mg/kg daily for 2 months, followed by L-NAME (25 mg/kg/day) for 1 week to inhibit all NO synthase isoforms. After treatment with L-NAME, we performed baseline renal function measurements, then infused L-arginine (2.5 mg/100 g BW x min) to reverse NO inhibition and assessed whether AG exerted NO-inhibitory activity independently of L-NAME. Prior to L-arginine infusion, AG-treated rats did not differ from controls with respect to body weight, kidney weight, systolic blood pressure, urine flow rate, urinary protein or albumin excretion, or urinary excretion of NO metabolites. After L-arginine infusion, all animals showed a 10-15% decrease in mean arterial blood pressure. L-Arginine-induced increases in urine flow, inulin clearance, PAH clearance, sodium excretion, and NO metabolite excretion were blunted in aminoguanidine-treated animals. To assess long-term effects of aminoguanidine, rats were treated for 12 months. Urinary excretion of NO metabolites was lower than controls. Inulin clearance was higher than controls. Aminoguanidine blunts the effect of L-ariginine on renal hemodynamics independently of the nitric oxide synthase inhibitor, L-NAME. However, the use of aminoguanidine for 12 months in rats did not adversely affect renal function. PMID- 9398026 TI - Glomerular basement membrane polyanionic sites and nitric oxide in genetically salt-sensitive and resistant hypertensive rats. AB - Cationic colloid gold, a polycationic histochemical probe, was used to analyze the distribution of glomerular basement membrane (GBM) polyanions, including heparan sulfate protoglycan in genetic salt-sensitive (SBH/Y) and resistant (SBN/Y) hypertensive rats, with or without high dietary salt intake. GBM morphology, renal function and nitric oxide, as measured by plasma and urine nitrite (NO2) and nitrate (NO3) were also determined. In the salt-sensitive rats the high-salt dietary intake resulted in severe hypertension, proteinuria and decreased glomerular filtration rate. After 1 month of high-salt intake, the average width of the GBM of salt-sensitive rats was higher by 27% than that of salt-resistant rats. The number of GBM anionic sites (lamina rata externa and interna) was much lower in both salt-sensitive and salt-resistant groups after 1 month of salt loading, 8.04+/-0.36 and 7.8+/-0.25 counts/cm, respectively, compared to the respective values of non-salt-loaded animals, 20.58+/-1.08 counts/cm in the SBH/Y (p < 0.001) and 21+/-1.86 counts/cm in the SBN/Y (p < 0.001). A decreased nitric oxide production was found in the salt-sensitive rats before and after salt loading compared with the salt-resistant group. No correlation was found between the nitric oxide changes and the GBM modifications. It is concluded that high-salt intake may be deleterious to the permselectivity of the GBM. It is suggested that salt restriction in hypertension may have a beneficial effect in preventing GBM permselectivity changes and proteinuria. PMID- 9398027 TI - Acute effects of angiotensin II receptor antagonist on autoregulation of zonal glomerular filtration rate in renovascular hypertensive rats. AB - This study was designed to assess the renal capability to autoregulate total blood flow, glomerular filtration rate (GFR) and local GFR in outer, middle and inner cortical layers (OC, MC, IC) in the two-kidney, one-clip (2K-1C) renovascular hypertensive rat, with or without acute infusion of the angiotensin II receptor antagonist losartan (5 mg/kg, i.v.). Age-matched, sham-operated Wistar rats were used as controls. The hemodynamic study in all animals was performed 4 weeks after clipping. The clipping increased blood pressure significantly, whereas losartan reduced the renal arterial pressure (RAP) from 165+/-8 to 125+/-6 mm Hg (p < 0.01) in 2K-1C hypertensive rats and reduced the RAP from 107+/-2 to 101+/-1 mm Hg (p < 0.05) in normotensive animals. Renal blood flow (RBF), total and local GFR were decreased in the nonclipped kidney of 2K-1C hypertensive rats compared with sham-operated rats, but losartan significantly increased the RBF and GFR. RBF was well maintained in response to reduction in RAP in the nonclipped kidneys with and without losartan treatment. The capability of total GFR autoregulation was impaired in untreated 2K-1C hypertensive rats and losartan-treated sham-operated rats, whereas losartan completely abolished GFR autoregulation in the nonclipped kidney of 2K-1C hypertensive rats. Losartan impaired autoregulation of zonal GFR to the same extent in all three cortical layers of sham-operated rats, whereas in the nonclipped kidney of 2K-1C hypertensive rats losartan had a more pronounced effect on the superficial GFR autoregulation than in middle and inner cortex, indicating that angiotensin II plays a major role in regulating the GFR response to the acute changes of renal arterial pressure. PMID- 9398029 TI - Microinjection studies on the effect of furosemide on the rat nephron. AB - The tubular site of furosemide (F) action was studied by the technique of diuretic microinjection (MIJ) into proximal tubules of the rat nephron. F was injected into the last proximal superficial loop of 51 proximal tubules, at the concentration of 3 x 10(-4) mol/l in an infusate that contained 14C-inulin. Collections were performed at the early distal tubule before and after MIJ. Single nephron filtration rate (SNGFR) remained unchanged, while the percent of filtered volume reabsorbed up to the site of collection was 85+/-2 before, 79+/ 2% after MIJ, p < 0.005. The calculated concentration of F in the collected distal tubular fluid during the post-MIJ measurements averaged 3 x 10(-5) mol/l. This experiment was repeated by injecting F into the early proximal convolutions of 43 nephrons, while the collections were performed at the last proximal segments. In these studies of proximal volume absorption, SNGFR was 34+/-3 before, 35+/-4 nl/min after F (p > 0.6). The respective percent reabsorptions were 70+/-3 and 73+/-3% (p +/- 0.3). In order to determine whether the technique per se was suitable to detect changes in reabsorption, the proximal MIJ study was repeated by using the carbonic anhydrase inhibitor dichlorphenamide 3 x 10(-5) mol/l in the microinjectate: while SNGFR remained unchanged, percent reabsorption fell from 63+/-5 to 45+/-7% during injection of the diuretic, p < 0.03. We conclude that the technique is adequate to examine the effects of drugs, and that F does not reduce proximal volume absorption at concentrations of 3 x 10(-5) mol/l. The loop diuretic decreases distal volume absorption by abolishing the osmotic gradient between blood and tubular fluid along the early distal convoluted tubule. PMID- 9398028 TI - Effect of intratubular application of angiotensin 1-7 on nephron function. AB - Cleavage of the C-terminal tripeptide of angiotensin I (Ang I) by neutral endopeptidase 24.11 releases angiotensin 1-7 (Ang 1-7). Because Ang I and neutral endopeptidase 24.11 are present in proximal tubular fluid and brush border, respectively, Ang 1-7 could be released into proximal tubular fluid to affect nephron function. Therefore, we studied the effect of intratubular Ang 1-7 (10( 12) to 10(-8) M) on nephron function employing in vivo renal micropuncture in inactin-anesthetized Munich-Wistar-Fromter rats. We observed that: (i) Intratubular application of Ang 1-7 for 3, 15, or 30 min did not affect reabsorption in the microperfused proximal convoluted tubule determined as net fluid reabsorption. (ii) During perfusion of Henle's loop for 15 min with artificial tubular fluid (time control), we observed a decline in fluid, potassium and sodium reabsorption by 20, 18 and 5%, respectively. A similar decline in reabsorption was seen with intratubular application of Ang 1-7 in a concentration of 10(-12) or 10(-10) M. In contrast, intratubular application of Ang 1-7 in a concentration of 10(-8) M increased fluid, potassium and sodium reabsorption in that nephron segment by 11, 9 and 3%, respectively. The latter response was completely abolished by AT1 angiotensin II receptor antagonist losartan (10[-6] M). (iii) Intratubular application of Ang 1-7 did not affect net sodium, potassium, or fluid reabsorption in the distal tubule. (iv) TGF response assessed by measuring proximal tubular stop-flow pressure or single nephron filtration rate during orthograde open-loop perfusion of Henle's loop was not significantly altered by intratubular application of Ang 1-7. These findings show that intratubular application of Ang 1-7 in concentrations which possibly cover the physiological range does not significantly alter (i) tubular reabsorption in proximal convoluted or distal tubule, or (ii) TGF response. Intratubular Ang 1-7 at a concentration of 10(-8) M appears to increase reabsorption in Henle's loop by an AT1 angiotensin II receptor-mediated mechanism, the physiological relevance of which remains to be established. PMID- 9398030 TI - Urinary calcium excretion and renal calbindin-D28k. AB - The present investigation examined the possible influence of urinary calcium excretion on the concentration of renal calbindin-D28k. Thiazide diuretics stimulate calcium transport across the epithelial cells of the distal tubule, which express calbindin-D28k in high concentrations. Calbindin-D28k is assumed to facilitate transcellular Ca diffusion. Reduced urine calcium excretion and increased urine output were induced in Wistar rats by infusion of bendroflume thiazide 1 mg/kg/day. The two control groups had infusions of either furosemide 20 mg/kg/day or vehicle, n = 8 in each group. Urinary Ca excretion was reduced to 10% in the thiazide group and increased by 50% in the furosemide group. Renal concentrations of calbindin-D28 showed no difference between vehicle, thiazide- and furosemide-treated rats. No differences in plasma concentrations of calcium, magnesium, phosphorus, urea, PTH, calcitonin and 1,25-(OH)2D were found between the groups. The present study describes that urine calcium excretion selectively can be manipulated without accompanying changes in renal calbindin-D28k concentrations. The data, therefore, suggest that urinary calcium excretion is not a significant determinator of cytosolic concentrations of renal calbindin D28k. PMID- 9398031 TI - Alterations of the renal handling of H+ in diabetic rats. AB - Renal acid excretion and proximal and distal nephron acidification were evaluated 20 days after induction of diabetes, in rats, by intraperitoneal injection of streptozotocin (45 mg/kg). Titratable acidity in urine was measured by microtitration and ammonium excretion (NH4+) by spectrophotometry. Proximal tubular acidification was evaluated by the kinetics of reabsorption of perfused HCO3-. Distal nephron acidification was evaluated by measuring urine - blood pCO2 differences under alkaline overload. The net acid excretion (titratable acidity + NH4+ - HCO3-) was higher (p < 0.001) in diabetic rats (9.82+/-0.65 micromol/min/kg, n = 26) than in the control group (6.34+/-0.14, n = 24). Proximal HCO3- reabsorption was also higher (p < 0.001) in diabetic rats (8.38+/ 0.11 nmol/cm2/s, n = 12) than in the control group (2.30+/-0.10, n = 22); however, evaluation of distal nephron H+ secretion by urine - blood pCO2 methodology was similar in both groups. We concluded that in rats with induced diabetes mellitus there is an increased rate of proximal HCO3- reabsorption, possibly effected by a higher density of Na+/H+ antiporter in the luminal membrane of the proximal tubule and by an increased proton-motive force of the H+ secretory mechanism. The higher rates of H+ secretion generate lower stationary proximal luminal pH and probably maintain the blood pH within the physiological range. PMID- 9398032 TI - Inhibition and restimulation by insulin of cellular autophagy in distal tubular cells of the kidney in early diabetic rats. AB - Cellular autophagy in the cells of distal tubular segments (DT cells) of the kidney cortex in streptozotocin-diabetic rats was evaluated by quantitative electron microscopy. Five days after streptozotocin administration, volume and numerical densities of autophagic vacuoles (AVs) in DT cells were reduced by 56 and 57%, respectively. The correction of the diabetic state by insulin injection reversed the inhibition of cellular autophagy. Volume and numerical densities of AVs increased by 97 and 53%, respectively. Endogenous insulin replacement by islet transplantation showed the same effect on cellular autophagy. Volume and numerical densities of AVs increased by 82 and 34%, respectively, as compared with sham-operated diabetic animals. The data show that, during diabetic kidney hypertrophy, the cellular autophagy is inhibited in DT cells to the same extent as in proximal tubular cells, suggesting that DT cells contribute in a balanced manner to hypertrophic growth of the kidney cortex. Similarly, DT cells are involved in the catabolic reaction, i.e., stimulation of autophagy, after metabolic correction of the diabetic state by insulin. PMID- 9398033 TI - Advantages of a two-chamber culture system to test drug nephrotoxicity: the example of cephaloridine. AB - Rabbit renal proximal tubular cells, cultured to confluency on a permeable collagen film in a two-chamber system, were exposed for 72 h to various concentrations of the nephrotoxic antibiotic, cephaloridine (CLD). A decrease in cellular proteins, leakage of lactate dehydrogenase and morphological changes appeared at CLD concentrations of 0.1, 1.0, and 0.5 mg/ml, respectively. The permeability of the monolayer to Lucifer yellow (LY), a dye that does not cross cell membranes, was increased by 1 or 2 mg/ml but not by lower concentrations of CLD. The large basolateral/apical glucose concentration gradient established by the cells was decreased by CLD. However, the fact that, at the CLD concentration of 1 mg/ml, LY totally equilibrated by diffusion across the monolayer, whereas the injured monolayer was still able to maintain a detectable glucose gradient, shows that damage caused by CLD to the diffusion barrier prevails over that affecting glucose uptake. Consistent with the data in the literature concerning the mechanism of CLD accumulation in renal cells, our results show that CLD was more toxic when it was added at the basolateral than at the apical side of the cultured cells. These results illustrate the advantages of using a two-chamber system of cell culture in nephrotoxicity studies. PMID- 9398034 TI - Renal changes induced by envenomation with Africanized bee venom in female Wistar rats. AB - Human envenomation caused by bee or wasp stings has been reported to cause acute renal failure (ARF), usually due to acute tubular necrosis (ATN), as a frequent complication. The pathogenetic mechanisms of ATN occurring in these accidents are still unclear. In the present study, female Wistar rats weighing 150-200 g were injected intravenously with Africanized bee venom at a dose of 0.4 microl/100 g body weight and used in functional and light microscopy studies. The animals were divided into two groups: the early group was studied 3-8 h after inoculation, and the late group was studied 24-30 h thereafter. The animals showed ARF characterized by reduction of glomerular filtration rate with increasing levels of plasma creatinine. They also showed increased fractional sodium and potassium excretions, suggesting changes in the proximal portion of the nephron. The water transport through collecting tubules was reduced, with consequent diuresis, indicating functional changes in the distal portion of the nephron. These functional changes were more marked in the early group, with recovery tending to occur after 24 h. Albuminuria was also observed in this group. Light microscopy showed ATN mainly in cortex and outer medulla, with isolated necrosis in cells or small groups of cells and cast formation in the distal and collecting tubules. After 24 h frequent mitotic figures were found in the tubular epithelium. The observed ARF was due to ATN which in turn was probably caused by multiple effects, mainly hemodynamic changes secondary to cardiotoxicity and systemic vasodilation caused by the venom, myohemoglobinuria, and the direct action of the venom on tubular cells. PMID- 9398035 TI - HPV detection in children prior to sexual debut. AB - Knowledge of the epidemiology of infection with human papillomavirus (HPV) in childhood is important, since HPV infection early in life could represent a risk factor for later development of anogenital cancer. A random sample of Danish children aged 0 to 17 years was tested for the presence of HPV in the anal region and the oral cavity by the polymerase chain reaction using a consensus HPV L1 primer. Only 4 of 249 anal beta-globin-positive samples and one of 392 oral beta globin-positive samples were HPV-positive. All HPV-positive samples were of unknown types. We conclude that the prevalence of anogenital HPV infection in childhood is very low indeed and that the oral cavity does not seem to act as a reservoir for HPV infection in childhood. This indicates that anogenital types of HPV are not transmitted to any measureable degree by non-sexual routes and further supports the notion that HPV infection takes place mainly later in life. PMID- 9398036 TI - Cancer incidence in female smokers: a 26-year follow-up. AB - A random sample of 26,000 Swedish women who were asked about their smoking habits in the early 1960s have now been followed for 26 years with respect to cancer incidence. Most findings regarding tobacco smoking and cancer from studies of men were confirmed also among the women. Elevated relative risk for current smokers compared with women who never smoked regularly were seen for cancers of the lung, upper aerodigestive sites, pancreas, bladder, cervix and all cancers combined, as well as a notably high relative risk for cancers of organs of the urinary tract other than kidney and bladder. Relative risk increased with dose, measured as grams of tobacco smoked per day, for cancers of the upper aerodigestive sites, lung, cervix, bladder, organs of the urinary tract other than kidney and bladder and all cancers combined. For cancers of the lung, bladder and cervix, there was an inverse relationship with age when starting to smoke tobacco. The reported inverse relationship between smoking and endometrial cancer could not be corroborated, nor was there any significant relationship between smoking and colorectal or breast cancer. PMID- 9398037 TI - Incidence of non-melanocytic skin cancer in Geraldton, Western Australia. AB - To measure the rate at which non-melanocytic skin cancers develop, we conducted a population-based, longitudinal study in Geraldton, Western Australia. Subjects were residents of Geraldton, Western Australia, who were between 40 and 64 years of age and registered on the electoral roll in 1987. In 1987 and again in 1992, dermatologists examined participants for skin cancers. They examined all skin areas, apart from those covered by underwear or hair. Subjects were asked about skin cancers that they had had treated between the 2 surveys. When all skin cancers were counted, the incidence rates of basal cell carcinoma were 3,379 per 100,000 person-years in women and 7,067 per 100,000 in men; those of squamous cell carcinoma were 501 per 100,000 in women and 775 per 100,000 in men. Sixteen percent of men and 14% of women developed at least one basal cell carcinoma; 2.8% of men and 2.2% of women had at least one squamous cell carcinoma. Most incident skin cancers were diagnosed at the second examination. More than half of the subjects who had a skin cancer at the first examination developed another. Squamous cell carcinomas occurred almost exclusively on parts of the body that are usually exposed. Basal cell carcinomas were common on the head, neck and trunk but not on the forearms and backs of hands. A quarter of people with a skin cancer on an exposed site also had one on the trunk. Our results show that skin cancer is extremely common in this population and frequently undiagnosed. Multiple skin cancers occur commonly, and skin cancers on exposed sites often are associated with skin cancers on less exposed sites. PMID- 9398038 TI - Dietary fat intake and risk of prostate cancer: a prospective study of 25,708 Norwegian men. AB - The relationship between incidence of prostate cancer and intake of dietary fat and foods rich in fat was studied in 25,708 men aged 16-56 years attending a Norwegian health screening in 1977-1983. Linkage to the Cancer Registry of Norway and the Central Bureau of Statistics of Norway ensured a complete follow-up until December 31, 1992. Diet was recorded on a semi-quantitative food-frequency questionnaire at the time of screening, and 72 cases of prostate cancer were identified during follow-up. At the end of follow-up, mean age of the total study sample was 56 years (range 19-68), while mean age at diagnosis of prostate cancer was 60 years (range 47-67). No association was found between energy-adjusted intake of total fat, saturated fat, mono-unsaturated fat or poly-unsaturated fat and the incidence of prostate cancer. Significant positive associations were found for body mass index (BMI) and consumption of hamburgers/meatballs, while no association was found with consumption of frankfurters/sausages and a significant negative association with the weekly number of main meals with meat. A significantly increased risk of prostate cancer was associated with skim milk as compared to whole milk. Milk preference (skim vs. whole) was associated significantly positively with BMI. Our study of a relatively young cohort does not confirm previous case-control and cohort studies suggesting that dietary fat, especially from animal sources, is associated positively with risk of prostate cancer. PMID- 9398039 TI - p16INK4, pRB, p53 and cyclin D1 expression and hypermethylation of CDKN2 gene in thymoma and thymic carcinoma. AB - There have been few reports on genetic alterations in thymomas. To investigate the expression of p16INK4A, RB, p53 and cyclin D1 in thymomas, we first examined 36 thymomas (non-invasive type, 16 cases; invasive type, 20 cases) and 3 thymic carcinomas, using immunohistochemistry. Abnormal expression of p16INK4A, RB, p53 and cyclin D1 was observed in 18, 8, 10 and 7 cases, respectively. Only a subgroup of invasive thymomas and thymic carcinomas showed an inverse correlation between p16INK4A and RB expression. Subsequently, we examined the 36 thymomas and 4 thymic carcinomas for mutations in p53 and CDKN2 genes, using PCR-SSCP and direct-sequencing analyses. No mutation of these genes was detected in the thymomas and thymic carcinomas examined. A polymorphism in the 3' untranslated region of exon 3 of CDKN2 was detected in 5 cases of thymoma. We searched for hypermethylation in the promoter region of CDKN2, observing it in 4 thymomas and 1 thymic carcinoma. Our data suggest that, unlike other more common cancers, alteration of the p53 gene may not play a significant role in the tumorigenesis of thymoma. However, inactivation of p16INK4A and RB may play a role in the progression of thymoma and thymic carcinoma. PMID- 9398040 TI - Non-Hodgkin's lymphoma among people with AIDS: incidence, presentation and public health burden. AIDS/Cancer Study Group. AB - We describe the anatomic and histologic presentation and prognosis of non Hodgkin's lymphoma (NHL) among people with AIDS (PWA) and determine their contribution to the NHL burden. We linked AIDS and cancer registries in selected areas of the United States and compared NHL sites and histologies in PWA and non PWA, after adjusting for age, sex and ethnicity. Among 51,033 PWA, we found 2,156 cases of NHL (4.3%). Half of NHL cases occurring post-AIDS were not reported to AIDS registries. NHL was part of an AIDS-defining condition for 3.2% of all PWA; the relative risk of NHL with 3.5 years of another AIDS diagnosis was 165-fold compared to non-PWA within the cancer surveillance system. Of NHLs, 39% were high grade (vs. 12% among non-PWA), 60% were nodal (vs. 74% among non-PWA) and 15% had brain primaries (vs. 1% among non-PWA). Excluding brain sites, extranodal sites were still 20% more common than expected. Relative risk was elevated for all histologic types, with the risk ranging from 652-fold for high-grade diffuse immunoblastic tumors and 261-fold for Burkitt's lymphomas to 113 for intermediate grade lymphoma to 14-fold for low-grade lymphoma. Survival among PWA with NHL was poor, and tumor grade had little impact. In high-risk AIDS areas, AIDS-related NHLs constitute a major share of the NHL burden. We conclude that NHL risk is considerably under-estimated in AIDS registry data. The major differences between PWA and non-PWA were the high frequency of brain lymphoma and the increase in high-grade lymphomas in PWA. However, the grade of NHL did not influence the prognosis among PWA with lymphoma. The increasing risk of NHL in PWA has contributed substantially to the general increase in NHL rates in the United States since 1981. PMID- 9398041 TI - Analysis of E2 amino acid variants of human papillomavirus types 16 and 18 and their associations with lesion grade and HLA DR/DQ type. AB - Human papillomavirus (HPV) 16 and HPV 18 E2 amino acid variants and host HLA DR/DQ haplotypes have been identified by direct nucleic acid sequencing from cervical scrapes. HPV16 E2 variants co-segregate with a nucleotide variant at nt350 (in E6), which previously has been associated with persistent infections. Both HPV16 and HPV18 E2 variants occur relatively more frequently in individuals with HLA DR/DQ haplotypes 0401/0301 and 1101/0301 but are not related to lesion grade. PMID- 9398042 TI - Reduced expression of the CXC chemokine hIRH/SDF-1alpha mRNA in hepatoma and digestive tract cancer. AB - We isolated human intercrine reduced in hepatomas (hIRH) as a mRNA whose expression was reduced in differential displays from human hepatocellular carcinoma. hIRH is equivalent to the alpha-chemokine SDF-1alpha/PBSF. We have previously demonstrated on Northern blot analysis that although hIRH mRNA expression is common in human normal tissues, it is absent from pre-malignant colonic adenomas and from 27 human malignant cell lines. However, there are no reports on the mRNA status of hIRH in other human cancers. The present study was designed to investigate semi-quantitatively the expression of hIRH/SDF-1alpha mRNA in hepatocellular carcinoma and digestive tract cancers by reverse transcription-polymerase chain reaction (RT-PCR). The expression of hIRH/SDF 1alpha in the majority of cancer tissues analyzed was markedly reduced compared with that in adjacent non-cancer tissue. RT-PCR was more sensitive than Northern blots in the detection of hIRH mRNA. The average (mean +/- SE) tumor/normal (T/N) ratio determined by RT-PCR was 0.40 +/- 0.07 in 10 pairs of hepatoma, 0.38 +/- 0.09 in 14 pairs of colon cancers, 0.43 +/- 0.07 in 10 pairs of esophageal cancers and 0.70 +/- 0.09 in 26 pairs of gastric cancers. As a control, the mean G3PDH T/N ratio was 1.16 +/- 0.06. The distribution of T/N ratios was significantly different between gastric cancer and the other cancers, but there was no correlation between hIRH/SDF-1alpha expression and clinicopathological characteristics in gastric cancer. Our findings demonstrate that hIRH/SDF-1alpha expression is reduced in the majority of gastrointestinal tumors. PMID- 9398043 TI - Increased presence of CD34+ cells in the peripheral blood of head and neck cancer patients and their differentiation into dendritic cells. AB - Patients with head and neck squamous cell carcinoma (HNSCC) have profound immune deficiencies. In 65% of these patients, there is an increased intra-tumoral presence of immune-suppressive CD34+ progenitor cells. The goal of the present study was to determine whether CD34+ cell levels were also increased in the peripheral blood of HNSCC patients and if these immune-suppressive cells could be differentiated into dendritic cells. Our results showed that CD34+ cell levels are increased in the peripheral blood of HNSCC patients. To assess if these CD34+ cells could differentiate into dendritic cells, they were isolated from the blood of HNSCC patients and cultured for 12 days with various cytokine combinations. Culturing CD34+ cells with stem cell factor (c-kit ligand) plus granulocyte macrophage colony-stimulating factor resulted in the appearance of a significant proportion of cells expressing phenotypic markers characteristic of dendritic cells. Also, including tumor necrosis factor-alpha yielded a significant proportion of cells resembling the bipotential precursor cells for dendritic cells and monocytes (CD14+CD1a+), in addition to the dendritic-like cells. When the differentiation inducer 1alpha,25-dihydroxyvitamin D3 [1,25(OH)2D3] was added along with the cytokine combinations, the yield of cells having characteristics of dendritic cells was further increased. Cells that were derived from CD34+ cell cultures containing 1,25(OH)2D3 had a more potent capacity to present the recall antigen tetanus toxoid to autologous peripheral blood leukocytes and to stimulate a mixed leukocyte response compared to cultures to which 1,25(OH)2D3 had not been added. Our results show that CD34+ cells, whose frequency is increased in HNSCC patients, can be differentiated into cells that phenotypically and functionally resemble dendritic cells and that 1,25(OH)2D3 accentuates this differentiation. PMID- 9398044 TI - Dietary fats and colon cancer: assessment of risk associated with specific fatty acids. AB - There are many biological mechanisms whereby dietary fat and specific dietary fatty acids may alter risk of colon cancer in addition to their contribution to total energy intake. To evaluate these potential associations, we used detailed dietary intake data collected in a population-based study of 1,993 incident colon cancer cases and 2,410 controls conducted in 3 areas of the United States. The most commonly consumed fatty acid in the study population was oleic acid. One third of dietary fats consumed came from additions to other foods at the table or from the preparation of other foods. After adjusting for total energy intake, physical activity and body size, neither total dietary fat nor specific fatty acids was associated with risk of colon cancer. However, among older women, fats from food preparation were associated with increased risk of colon cancer (OR 1.84, 95% CI 1.20-2.80), while fats from foods themselves or from additions to other foods were not. While dietary fats were not associated with colon cancer risk in the total population, subgroups of the population appeared to be at slightly greater risk if they consumed a high-fat diet. Women who consumed a diet high in mono-unsaturated fatty acids (MFAs) and poly-unsaturated fatty acids (PFAs) and who had a family history of colorectal cancer were at greater risk of colon cancer than those with similar intakes but without a family history of colorectal cancer. Similar associations with family history were noted among men diagnosed at younger ages for MFA, linolenic acid and 20-carbon PFA. PMID- 9398045 TI - Phenotypic analysis of ovarian carcinoma: polypeptide expression in benign, borderline and malignant tumors. AB - Studies of multiple markers in tumors are required for adequate biological characterization. We have characterized the expression of multiple proteins in human ovarian tumors using the technique of 2-dimensional gel electrophoresis (2 DE/PDQUEST). Tumor cells were prepared from the tissue of 22 ovarian tumors. Large variations were observed between tumors in the expression of various polypeptides, indicating heterogeneity in gene expression. An increase in the spot density of 2 cell-cycle-related proteins, PCNA and OP18/stathmin, was observed in carcinomas. Borderline tumors expressed low levels of these proteins. Significant increases in the levels of nm23, GST-pi, elongation factor 2 and triose phosphate isomerase were recorded in ovarian carcinomas. Furthermore, decreases in the levels of tropomyosin-2 and lamin C were observed in malignant as compared with benign tumors. The pattern of expression of 9 protein markers was examined in individual tumors. All malignant tumors showed simultaneous alterations in the expression of 5 or more of these proteins, whereas no benign tumor showed alterations in the expression of more than 3 polypeptides. Borderline tumors showed alterations in 0 to 6 markers. We conclude that the simultaneous analysis of multiple polypeptides, which can be achieved by 2-DE, is useful for characterization of gene expression and diagnostic studies in ovarian tumors. PMID- 9398046 TI - Gastric cancer, gastritis and plasma vitamin C: results from an international correlation and cross-sectional study. The Eurogast Study Group. AB - Low intake of foods rich in vitamin C is associated with an increased risk of gastric cancer, and geographic variation in average vitamin C intake, therefore, could explain some of the wide international variation in gastric cancer rates. This multicentre study investigated the relationships between plasma levels of vitamin C, as an indicator of vitamin C intake, and gastric cancer rates, markers of gastritis and other socio-demographic variables. Fasting plasma samples from about 1,400 individuals from 9 centres in 7 countries worldwide were assayed for total vitamin C using a fluorometric assay. There was no association between average plasma vitamin C levels and either gastric cancer mortality or incidence rates in the populations studied. Therefore, variation in fasting plasma vitamin C levels, as an indicator of consumption of vitamin C, does not appear to explain any of the wide geographic variation in gastric cancer rates. Furthermore, there was no association between plasma vitamin C levels and Helicobacter pylori infection, low serum levels of pepsinogen A (as a marker of severe chronic atrophic gastritis) or the presence of DNA adducts in blood leukocyte DNA. Multivariate models showed that fasting plasma vitamin C levels were associated positively with female sex, higher levels of education, never having smoked and increasing height and negatively with number of cigarettes smoked per day and increasing weight. This suggests not only that gender and tobacco smoking, in particular, are important predictors of plasma vitamin C levels but also that their effects are consistent throughout the developed world. PMID- 9398047 TI - Investigation of mammary epithelial cell-bone marrow stroma interactions using primary human cell culture as a model of metastasis. AB - A model has been established using primary human cell culture to study the cell biology of breast cancer metastasis to bone marrow. Mammary epithelia were obtained in single cell suspension from tumour (macroscopically involved), benign (macroscopically uninvolved) and normal (reduction mammoplasty) breast tissue as well as from locally involved lymph nodes. Stromal layers were generated from long-term cultures of human bone marrow or from mammary fibroblasts derived from normal or malignant tissue. The interaction between epithelia and stroma has been studied in terms of adhesion of the epithelia to the stroma and their subsequent growth in co-culture. Our results show that when assayed up to 9 hr after plating, epithelial cells from malignant tissue (14 primary tumours and 9 metastases in lymph nodes) displayed a significant preference for adhesion to bone marrow stroma compared with mammary fibroblasts. In contrast, epithelial cells from 4 normal and 2 of 4 benign samples showed no significant preferential adherence. Subsequent co-culture of mammary epithelia with each of the 3 stromal layers revealed that under serum-free, in vitro conditions, bone marrow stromal layers did not provide an advantageous environment for colony growth, in contrast to their ability to provide a preferential substratum for adhesion. PMID- 9398048 TI - Cell-transforming activity and genotoxicity of phenolphthalein in cultured Syrian hamster embryo cells. AB - Phenolphthalein is a cathartic agent widely used in non-prescription laxatives. For the simultaneous assessment of in vitro carcinogenicity and mutagenicity of phenolphthalein, the ability of this chemical to induce cell transformation and genetic effects was examined using the Syrian hamster embryo (SHE) cell model. Cell growth was reduced by treatment with phenolphthalein at 10-40 microM in a dose-related manner. Treatment with phenolphthalein for 48 hr induced a dose dependent increase in morphological transformation of SHE cells. Over the dose range that resulted in cell transformation ( 10-40 microM), treatment of SHE cells with phenolphthalein induced gene mutations at the hprt locus but not at the Na+/K+ ATPase locus. A statistically significant level of chromosomal aberrations was elicited in SHE cells treated with phenolphthalein at the highest dose (40 microM). Meanwhile, neither numerical chromosomal changes nor DNA adduct formation, analyzed by the nuclease P1 enhancement version of 32P-post-labeling, were induced by treatment with phenolphthalein at any concentrations examined. We thus report cell-transforming activity and mutagenicity of phenolphthalein assessed with the same mammalian cells in culture. Our results provide evidence that phenolphthalein has cell-transforming and genotoxic activity in cultured mammalian cells. The mutagenic and clastogenic activities of phenolphthalein could be a causal mechanism for carcinogenicity in rodents. PMID- 9398049 TI - Modulation of p53 expression in cultured colonic adenoma cell lines by the naturally occurring lumenal factors butyrate and deoxycholate. AB - The high incidence of colorectal cancer in Western society is believed to be strongly related to diet. Mutation of the p53 gene is a late event in colorectal carcinogenesis, and thus, the majority of pre-malignant adenomas express wild type p53. As loss of p53 protein function is an important step in colorectal carcinogenesis, we investigated whether naturally occurring lumenal factors can modulate the expression of p53 in non-tumorigenic human colonic adenoma cell lines. Levels of p53 protein and mRNA were measured in adherent cells which had been incubated with growth-inhibitory concentrations of sodium butyrate (a by product of dietary fibre fermentation) or sodium deoxycholate (a bile acid) for up to 48 hr. We report that both butyrate and deoxycholate can down-regulate the expression of wild-type and mutant p53. In contrast, incubation for 48 hr with the endogenous inhibitory growth factor TGFbeta1 did not alter p53 protein expression. Thus, in addition to cellular mechanisms which regulate p53 function, such as post-translational stabilisation, nuclear exclusion, negative feedback inhibition of p53 mRNA translation or binding of p53 by cellular proteins, p53 protein levels also may be regulated by changes in the level of p53 gene transcription. Furthermore, we show that lumenal factors are able to affect directly the expression of p53 protein in colonic epithelial cells. PMID- 9398050 TI - Topoisomerase-I inhibitors for human malignant glioma: differential modulation of p53, p21, bax and bcl-2 expression and of CD95-mediated apoptosis by camptothecin and beta-lapachone. AB - Beta-lapachone and camptothecin are structurally unrelated agents thought to inhibit topoisomerase-I activity through distinct mechanisms. We find that beta lapachone is much more potent than camptothecin in inducing acute cytotoxic effects on human malignant glioma cells. Acute cytotoxicity induced by both drugs is apoptotic by electron microscopy, but not blocked by inhibitors of RNA or protein synthesis and not associated with changes in the expression of bcl-2, bax, p53, p21 or GADD45 proteins. In contrast, prolonged exposure of glioma cells to both drugs for 72 hr results in growth inhibition and apoptosis, with EC50 values around 1 microM. None of 7 glioma cell lines tested were resistant to either drug. LN-229 cells which have partial p53-wild-type activity show enhanced expression of p53, p21 and bax protein, whereas bcl-2 levels decrease, after exposure to camptothecin. In contrast, beta-lapachone increases bax protein expression in the absence of p53 activation. T98G cells are mutant for p53. In these cells, p53 levels do not change and p21 is not induced. bax accumulation in T98G cells is induced by both drugs, with bcl-2 levels unaltered. Surprisingly, ectopic expression of murine bcl-2 fails to abrogate the toxicity of either drug. Camptothecin, but not beta-lapachone, sensitizes human malignant glioma cells to apoptosis induced by the cytotoxic cytokines, tumor necrosis factor-alpha and CD95 ligand. Thus, both drugs have potent anti-glioma activity that may be mediated by enhanced bax expression but is not inhibited by ectopic bcl-2 expression. Camptothecin-like agents are particularly promising for immunochemotherapy of malignant glioma using cytotoxic drugs and CD95 ligand. PMID- 9398051 TI - Abundance of BRCA1 transcripts in human cancer and lymphoblastoid cell lines carrying BRCA1 germ-line alterations. AB - A competitive polymerase chain reaction has been developed for quantitation of BRCA1 mRNA. In human cancer cell lines, the amount of BRCA1 mRNA is relatively low, ranging from 6 to 38 copies per cell. The decay rate of these transcripts in actinomycin-treated cells indicates that the half-life of these molecules is about 4 hr, suggesting that the low concentration of BRCA1 messages is not due to molecular unstability. In human lymphoblastoid cell lines derived from patients carrying germ-line alterations of BRCA1, the amount of BRCA1 mRNA per cell is lowered only in cell lines exhibiting alterations leading to specific loss of transcripts from the mutated allele. These data indicate that the amount of BRCA1 available in these cells can be related directly to the number of "active" allele. PMID- 9398052 TI - Chemopreventive effects of diosmin and hesperidin on N-butyl-N-(4 hydroxybutyl)nitrosamine-induced urinary-bladder carcinogenesis in male ICR mice. AB - The chemopreventive effects of 2 flavonoids (diosmin and hesperidin) on N-butyl-N (4-hydroxybutyl)nitrosamine (OH-BBN)-induced urinary-bladder carcinogenesis were examined in male ICR mice. Animals were divided into 11 groups, and groups 1 to 7 were given OH-BBN (500 ppm) in the drinking water for 6 weeks. Groups 2 to 4 were fed diets containing the test compounds (group 2, 1000 ppm diosmin; group 3, 1000 ppm hesperidin; group 4,900 ppm diosmin + 100 ppm hesperidin) for 8 weeks during the initiation phase, while groups 5 to 7 were fed these diets, respectively, for 24 weeks during the post-initiation phase. Groups 8 to 11 were controls, given only the test compounds or untreated basal diets throughout the experiment (weeks 1 to 32). The incidence of bladder lesions and cell-proliferation activity estimated by enumeration of silver-stained nucleolar-organizer-region-associated proteins (AgNORs) and by the 5-bromodeoxyuridine (BUdR)-labeling index was compared among the groups. Feeding of the test compounds, singly or in combination, during both phases caused a significant reduction in the frequency of bladder carcinoma and preneoplasia. Dietary administration of these compounds significantly decreased the AgNOR count and the BUdR-labeling index of various bladder lesions. These findings suggest that the flavonoids diosmin and hesperidin, individually and in combination, are effective in inhibiting chemical carcinogenesis of the bladder, and that such inhibition might be partly related to suppression of cell proliferation. PMID- 9398053 TI - Intracellular Ca2+ release mediates ursolic acid-induced apoptosis in human leukemic HL-60 cells. AB - The effect of ursolic acid (UA) on tumor cell apoptosis was investigated using HL 60 human promyelocytic leukemia cells as a model cellular system. Treatment with UA resulted in a concentration-dependent decreased cell viability assessed by MTT assay. UA also induced genomic DNA fragmentation, a hallmark of apoptosis, indicating that the mechanism by which UA induced cell death was through apoptosis. The intracellular Ca2+ level was increased by treatment with UA. Intracellular Ca2+ inhibitors, such as intracellular Ca2+-release blockers (dantrolene, TMB-8 and ruthenium red) and an intracellular Ca2+ chelator (BAPTA/AM), significantly blocked the UA-induced increased intracellular Ca+ concentration. These inhibitors also blocked the effects of UA on cell viability and apoptosis. These results suggest that enhanced intracellular Ca2+ signals may be involved in UA-induced apoptosis in HL-60 cells. PMID- 9398054 TI - Sequence-dependent activity of the irinotecan-5FU combination in human colon cancer model HT-29 in vitro and in vivo. AB - Irinotecan, a DNA-topoisomerase-I inhibitor, is active against metastatic colon carcinoma. We investigated the effects of irinotecan and 5FU combinations in human colon-carcinoma cell line HT-29, both in vitro and in vivo. Cytotoxicity of 24-hr exposure was evaluated by SRB technique and the nature of interactions were determined by median-effect analysis. Strong synergism between irinotecan and 5FU was observed after sequential exposure, and only additivity after simultaneous exposure. At 50% level of kill, the mean sums of fractional effects were 0.13 +/- 0.05 and 0.18 +/- 0.02 respectively for the 2 sequential schedules, indicating that the combined amount of the 2 drugs necessary to kill 50% cells was only 0.18 and 0.13 times respectively, as much as would be required if they demonstrated purely additive behavior. In nude-mice xenografts, schedule-dependent toxicity was observed: the schedule in which irinotecan was administered i.v. 6 hours before 5FU was the most toxic. Higher anti-tumoral activity was noted when 20 mg/kg/day of each drug was administered sequentially (a delay of 6 hr between the 2 drugs) to mice over 5 days, in comparison with simultaneous administration. In vivo data confirmed those obtained in vitro in the same human colon-cancer model. These results suggest that irinotecan and 5FU combinations are of clinical interest and that the schedule of administration is a critical parameter for chemotherapeutic efficacy. PMID- 9398055 TI - Long-term inhibitory effects of a novel anti-estrogen on the growth of ZR-75-1 and MCF-7 human breast cancer tumors in nude mice. AB - The effects of the novel anti-estrogen EM-343 on the growth of 2 hormone responsive human breast cancer tumors have been examined in athymic nude mice. At the low daily dose of 5 microg, EM-343 administered subcutaneously for 6 months completely blocked the stimulatory effect of endogenous estrogens on the growth of ZR-75-1 and MCF-7 tumors implanted in nude mice. In addition, uterine weight decreased by 60% while ovarian weight increased by 37%. Estrogen receptor (ER) levels measured by [3H]-labeled estrogen binding were markedly reduced (by 96%, 96% and 92%) in ZR-75-1 and MCF-7 tumors, and in the mouse uterus, respectively. Accompanying the decrease in ER, progesterone receptor levels were reduced by 79%, 87% and 76%, respectively, in the above-mentioned tissues following EM-343 treatment. Our data show the pure anti-estrogenic properties of EM-343 and its high potency as an inhibitor of growth of human ZR-75-1 and MCF-7 breast tumors in nude mice. PMID- 9398056 TI - Retinoic acid-enhanced invasion through reconstituted basement membrane by human SK-N-SH neuroblastoma cells involves membrane-associated tissue-type plasminogen activator. AB - Al-trans retinoic acid (RA) enhanced human, S-type, SK-N-SH neuroblastoma cell invasion of reconstituted basement membrane in vitro but did not induce terminal differentiation of this cell line. In contrast to basal invasion, which was urokinase (uPA)- and plasmin-dependent, RA-enhanced invasion was dependent on tissue-type plasminogen activator (t-PA) and plasmin activity. Neither basal nor RA-enhanced invasion involved TIMP-2 inhibitable metalloproteinases. Enhanced invasion was associated with the induction of t-PA expression, increased expression of the putative t-PA receptor amphoterin, increased association of t PA with cell membranes and increased net membrane-associated PA activity. Enhanced invasion was not associated with significant changes in the expression of uPA or its membrane receptor UPAR; plasminogen activator inhibitors PAI-1 and PAI-2; metalloproteinases MMP-1, MMP-2, MMP-3, MMP-9 and membrane type MMP1; or tissue inhibitors of metalloproteinases TIMP-1 and TIMP-2. RA stimulated the association of t-PA with the external cell membrane surface, which could be inhibited by heparin sulphate but not by mannose sugars or chelators of divalent cations, consistent with a role for amphoterin. Our data indicate that RA can promote the malignant behavior of S-type neuroblastoma cells refractory to RA mediated terminal differentiation by enhancing their basement membrane invasive capacity. We suggest that this results from the action of a novel, RA-regulated mechanism involving stimulation of t-PA expression and its association with the cell membrane leading to increased PA-dependent matrix degradation. PMID- 9398057 TI - In vivo effects of activated H-ras oncogene expressed in the liver and in urogenital tissues. AB - Transgenic mouse technology provides a direct genetic approach to in vivo carcinogenesis. In order to determine the oncogenic potential of an activated ras gene in liver, kidney and intestine, we created transgenic mice expressing the human H-ras oncogene under control of the L-type pyruvate-kinase gene. This gene is expressed in hepatocytes, enterocytes, proximal tubular cells of the kidney and endocrine pancreatic cells. Depending on lines, we observed hepatocarcinoma, polycystic kidney disease and an unexpected epididymis hyperplasia. These transgenic mice are an interesting model of polycystic kidney disease, and complete our study of the tissue-specificity of oncogene action. PMID- 9398058 TI - Up-regulation of Fas (CD95) in human p53wild-type cancer cells treated with ionizing radiation. AB - Fas is a cell-surface protein which belongs to the tumor-necrosis-factor-receptor family. Signals through Fas are able to induce apoptosis in sensitive cells, and thus modalities for regulating the level of Fas expression on tumor cells are needed. We have studied cellular responses to gamma irradiation. The level of p53 tumor-suppressor protein was found to be elevated 3 hr after irradiation of p53wild-type MCF-7 breast-carcinoma cells. Interestingly, accumulation of p53 was followed by up-regulation of surface Fas levels between 4 and 8 hr after irradiation. The level of Fas up-regulation was dependent on dose and, whereas elevation in the level of p53 was transient, enhancement of Fas expression was stable. Fas up-regulation occurred coincidentally with induction of G1 cell-cycle arrest, a post-irradiation phenomenon known to be dependent on wild-type-p53 activity. We studied 9 other tumor lines, 2 with wild-type p53, 5 with mutant p53, and 2 expressing no p53. All lines expressing wild-type p53 were found to arrest in G1 and to up-regulate Fas after irradiation. In contrast, all 7 p53null and p53mutant lines failed not only to arrest their cell cycles in G1 phase, but also to up-regulate Fas levels in response to treatment. These findings demonstrate a direct correlation between wild-type-p53 activity and Fas up regulation after treatment with ionizing radiation, strongly suggesting that post irradiation Fas up-regulation is dependent on wild-type-p53 activity. Since low doses of radiation were sufficient to modulate Fas expression, up-regulation of the Fas death receptor may have clinical implications following radiotherapy. PMID- 9398059 TI - Characterisation of novel human lung carcinoma cell lines selected for resistance to anti-neoplastic analogues of staurosporine. AB - The staurosporine analogues CGP 41251, UCN-01 and Ro 31-8220 are specific inhibitors of protein kinase C (PKC). CGP 41251 and UCN-01 exert anti-neoplastic activity against human tumours grown in rodents, and CGP 41251 reverses multidrug resistance. The hypothesis was tested that these agents can induce drug resistance and alter cellular levels of target kinases. Human-derived A549 lung carcinoma cells were exposed for 6 months to CGP 41251, UCN-01 or Ro 31-8220 at gradually increasing concentrations. Cells acquired resistance against these agents, 4.3-fold against CGP 41251 (A549/CGP cells), 4.0-fold against UCN-01 (A549/UCN cells) and 14-fold against Ro 31-8220 (A549/Ro cells). Cells were neither collaterally cross-resistant towards the PKC inhibitors nor resistant against the growth-inhibitory properties of 12-O-tetradecanoylphorbol-13-acetate. However, cross-resistance was observed in A549/CGP cells against staurosporine (13-fold) and in A549/Ro cells against doxorubicin (26-fold). All 3 cell types expressed multidrug resistance-associated protein, and A549/Ro cells expressed P glycoprotein, as adjudged by Western blot analysis. Phorbol ester-stimulated PKC activity in these cells was decreased by between 57% and 96% compared to wild type A549 cells. Levels of the PKC isoenzymes alpha and theta in all 3 resistant cell types and of PKC-epsilon in A549/UCN cells were concomitantly reduced. Cells regained drug sensitivity after culture in the absence of drug for 6 (A549/Ro cells), 5 (A549/CGP cells) and 1 (A549/UCN cells) months. Our results suggest the following features of this type of anti-signalling drug: (i) they can induce drug resistance, (ii) they may be potentially useful in combination because of the lack of cross-resistance between them and (iii) they can down-regulate PKC, which may have pharmacological or toxicological consequences. PMID- 9398060 TI - Melanoma progression-associated glycoprotein MUC18/MCAM mediates homotypic cell adhesion through interaction with a heterophilic ligand. AB - MUC18/MCAM is a cell-surface glycoprotein that is strongly expressed on advanced human melanomas. Transfection of 3 MCAM-negative melanoma cell lines with MCAM cDNA led to cell-surface expression and to a MCAM-dependent homotypic adhesion. This adhesion was independent of divalent cations and was inhibited at 4 degrees C. Mixed aggregation assays with MCAM-expressing and non-expressing cells revealed that MCAM can function as a heterophilic cell adhesion molecule interacting with a non-MCAM ligand. Although MCAM contains a potential glycosaminoglycan-binding site, cell-surface glycosaminoglycans do not appear to be involved in the heterophilic adhesion observed here since these molecules were not able to influence the adhesion. Using a functional adhesion assay, 4/4 melanoma cell lines examined were found to express an MCAM ligand. In contrast, no evidence for an MCAM ligand was found on the 2 carcinoma or 2 hematopoietic cell lines examined. Stable transfection of an MCAM ligand-negative colorectal cell line resulted in MCAM surface expression but not in homotypic adhesion, indicating that homophilic MCAM-MCAM adhesive interactions may not occur. Our results suggest that MCAM expression by melanoma cells is associated with increased homotypic adhesion, an event that may support tumor cell survival and growth in vivo. PMID- 9398061 TI - Human papillomavirus types 52 and 58 are prevalent in cervical cancer from Chinese women. PMID- 9398063 TI - Advances in apoptosis research. AB - Apoptosis, also called programmed cell death, has attracted great attention in recent years. After its discovery by Carl Vogt in 1842, apoptosis research was dormant for more than a century. Its rediscovery in the second half of this century, and the coining of the term apoptosis in 1972 by Kerr, Wyllie, and Currie, ignited an unparalleled interest in this field of science. The number of publications related to apoptosis has been growing exponentially every year ever since. This is mainly due to three major advances, two of which have been made recently and one that is currently seen. First, studies with the small nematode Caenorhabditis elegans have identified a number of apoptosis regulating genes- the first evidence that cell death is an active process under genetic control. Many of these genes have mammalian homologs that, like their worm counterparts, seem to regulate mammalian apoptosis. Second, elucidation of the signal transduction pathways of apoptosis has lead especially to the identification of specific death signaling molecules such as a new family of cysteine proteases, the caspases. Third, it has now become clear that many diseases are characterized by dysregulation of apoptotic programs. Many of these programs involve a family of receptors and their ligands, the death receptor/ligand family. The hope now is to interfere with apoptosis regulation in these systems and to develop new therapeutic concepts. PMID- 9398064 TI - Encoding strategies in combinatorial chemistry. PMID- 9398065 TI - Neural codes: firing rates and beyond. AB - Computational neuroscience has contributed significantly to our understanding of higher brain function by combining experimental neurobiology, psychophysics, modeling, and mathematical analysis. This article reviews recent advances in a key area: neural coding and information processing. It is shown that synapses are capable of supporting computations based on highly structured temporal codes. Such codes could provide a substrate for unambiguous representations of complex stimuli and be used to solve difficult cognitive tasks, such as the binding problem. Unsupervised learning rules could generate the circuitry required for precise temporal codes. Together, these results indicate that neural systems perform a rich repertoire of computations based on action potential timing. PMID- 9398067 TI - Gene therapy. AB - In recent years, there have been a number of technological breakthroughs that have allowed for clinical trials in gene therapy to be initiated. In combination with the genome initiative, the potential for new therapeutics is limitless. Although an enormous amount of information has been obtained in a relatively short period of time, gene therapy is not yet ready for wide-scale practice. Some of the successes and obstacles that remain are summarized in this report. PMID- 9398068 TI - Motor proteins of the eukaryotic cytoskeleton. PMID- 9398069 TI - Scanning-probe-based science and technology. PMID- 9398070 TI - Fun with genealogy. PMID- 9398071 TI - Life without DNA repair. PMID- 9398072 TI - Origins of mRNA identity: capping enzymes bind to the phosphorylated C-terminal domain of RNA polymerase II. PMID- 9398074 TI - The orange carotenoid protein of Synechocystis PCC 6803. AB - A water soluble protein with the carotenoid 3'-hydroxyequinenone bound to it has been purified from the cyanobacterium Synechocystis PCC 6803. Based on partial amino acid sequencing of the protein, oligonucleotides were synthesized and used as primers for PCR to obtain a substantial fragment of the gene. This DNA was sequenced and the sequence data and the size of the protein indicate that the protein is encoded by gene slr 1963 in the Kazusa DNA sequence data bank containing the Synechocystis 6803 genome. This protein is very similar to 3' hydroxyechinenone proteins found in several other cyanobacteria but it shows very little resemblance in its amino acid or gene sequence to other carotenoid binding proteins. The protein binds 1-2 molecules of 3'-hydroxyechinenone and is slowly cleaved by proteases in the cell extract to give a molecule of approximately half the original mass which retains the carotenoid and which shows a striking change in color. PMID- 9398075 TI - Contribution to the physiological characterization of glycerol active uptake in Saccharomyces cerevisiae. AB - Evidence is presented here that in Saccharomyces cerevisiae IGC 3507, grown either on glycerol, ethanol or acetate, glycerol is transported by a high affinity uptake system of the electrogenic proton symport type, with Km of 1.7 +/ 0.7 mM, Vmax 441 +/- 19 micromolh(-1) g(-1) dry weight and a stoichiometry of 1:1 proton per molecule of glycerol, at 30 degrees C and pH 5.0. No competitors were found among other polyols and sugars. Glycerol maximum accumulation ratios followed p.m.f. with extracellular pH. CCCP prevented glycerol accumulation, and inhibited uptake. NaCl did not interfere with H+/glycerol kinetics and energetics. This transport system was shown to be under glucose repression and inactivation. Glucose-grown cells presented, instead, a lower affinity permease for glycerol, probably a facilitated diffusion. Growth on glucose in the presence of NaCl did not induce the high affinity carrier. The stringent control of cell physiological condition over induction suggests for glycerol proton symport rather a physiological role connected with growth under gluconeogenic conditions. PMID- 9398076 TI - The pH dependences of reactions catalyzed by the complete proton-translocating transhydrogenase from Rhodospirillum rubrum, and by the complex formed from its recombinant nucleotide-binding domains. AB - Transhydrogenase couples the translocation of protons across a membrane to the transfer of reducing equivalents between NAD(H) and NADP(H). Using transhydrogenase from Rhodospirillum rubrum we have examined the pH dependences of the 'forward' and 'reverse' reactions, and of the 'cyclic' reaction (NADP(H) dependent reduction of the analogue, acetyl pyridine adenine dinucleotide, by NADH). In the case of the membrane-bound protein in chromatophores, the imposition of a protonmotive force through the action of the light-driven electron-transport system, stimulated forward transhydrogenation, inhibited reverse transhydrogenation, but had no effect on the cyclic reaction. The differential response at a range of pH values provides evidence that hydride transfer per se is not coupled to proton translocation and supports the view that energy transduction occurs at the level of NADP(H) binding. Chromatophore transhydrogenase and the detergent-dispersed enzyme both have bell-shaped pH dependences for forward and reverse transhydrogenation. The cyclic reaction, however, is rapid at low and neutral pH, and is attenuated only at high pH. A mixture of recombinant purified NAD(H)-binding domain I, and NADP(H)-binding domain III, of R. rubrum transhydrogenase carry out the cyclic reaction with a similar pH profile to that of the complete enzyme, but the forward and reverse reactions were much less pH dependent. The rates of release of NADP+ and of NADPH from isolated domain III were pH independent. The results are consistent with a model for transhydrogenation, in which proton binding from one side of the membrane is consequent upon the binding of NADP+ to the enzyme, and then proton release on the other side of the membrane precedes NADPH release. PMID- 9398077 TI - Exchangeability of the b subunit of the Cl(-)-translocating ATPase of Acetabularia acetabulum with the beta subunit of E. coli F1-ATPase: construction of the chimeric beta subunits and complementation studies. AB - The gene encoding the b subunit of the Cl(-)-translocating ATPase (aclB) was isolated from total RNA and poly(A)+ RNA of Acetabularia acetabulum and sequenced (total nucleotides of 3038 bp and an open reading frame with 478 amino acids). The deduced amino acid sequence showed high similarity to the beta subunit of the F type ATPases, but was different in the N-terminal 120 amino acids. The role of the N-terminal region was investigated using an F -ATPase beta-less mutant of E. coli, JP17. The JP17 strain expressing the aclB could not grow under conditions permitting oxidative phosphorylation, although ACLB was detected in the membrane fraction. The beta subunit was divided into three portions: amino acid position from 1 to 95 (portion A), 96 to 161 (portion B) and 162 to the C-terminus (portion C). The corresponding regions of ACLB were designated as portions A' (from 1 to 106), B' (from 107 to 172) and C' (from 173 to 478). Chimeric proteins with combinations of A-B'-C', A-B-C' and A'-B-C restored the function as the beta subunit in E. coli F0F1-complex, but those with combinations of A'-B'-C and A-B' C had no function as the beta subunit. These findings suggested that portion B plays an important role in the assembly and function of the beta subunit in the F0F1-complex, while portion B' of ACLB exhibited inhibitory effects on assembly and function. In addition, portion A was also important for interaction of the beta subunit with the alpha subunit in E. coli F0F1-complex. These findings also suggested that the b subunit of the Cl(-)-translocating ATPase of A. acetabulum has a different function in the Cl(-)-translocating ATPase complex, although the primary structure resembled to the beta subunit of the F1-ATPase. PMID- 9398078 TI - Study of regulation of mitochondrial respiration in vivo. An analysis of influence of ADP diffusion and possible role of cytoskeleton. AB - The purpose of this work was to investigate the mechanism of regulation of mitochondrial respiration in vivo in different muscles of normal rat and mice, and in transgenic mice deficient in desmin. Skinned fiber technique was used to study the mitochondrial respiration in the cells in vivo in the heart, soleus and white gastrocnemius skeletal muscles of these animals. Also, cardiomyocytes were isolated from the normal rat heart, permeabilized by saponin and the "ghost" (phantom) cardiomyocytes were produced by extraction of myosin with 800 mM KCl. Use of confocal immunofluorescent microscopy and anti-desmin antibodies showed good preservation of mitochondria and cytoskeletal system in these phantom cells. Kinetics of respiration regulation by ADP was also studied in these cells in detail before and after binding of anti-desmine antibodies with intermediate filaments. In skinned cardiac or soleus skeletal muscle fibers but not in fibers from fast twitch skeletal muscle the kinetics of mitochondrial respiration regulation by ADP was characterized by very high apparent Km (low affinity) equal to 300-400 microM, exceeding that for isolated mitochondria by factor of 25. In skinned fibers from m. soleus, partial inhibition of respiration by NaN3 did not decrease the apparent Km for ADP significantly, this excluding the possible explanation of low apparent affinity of mitochondria to ADP in these cells by its rapid consumption due to high oxidative activity and by intracellular diffusion problems. However, short treatment of fibers with trypsin decreased this constant value to 40-70 microM, confirming the earlier proposition that mitochondrial sensitivity to ADP in vivo is controlled by some cytoplasmic protein. Phantom cardiomyocytes which contain mostly mitochondria and cytoskeleton and retain the normal shape, showed also high apparent Km values for ADP. Therefore, they are probably the most suitable system for studies of cellular factors which control mitochondrial function in the cells in vivo. In these phantom cells anti-desmin antibodies did not change the kinetics of respiration regulation by ADP. However, in skinned fibers from the heart and m. soleus of transgenic desmin-deficient mice some changes in kinetics of respiration regulation by ADP were observed: in these fibers two populations of mitochondria were observed, one with usually high apparent Km for ADP and the second one with very low apparent Km for ADP. Morphological observations by electron microscopy confirmed the existence of two distinct cellular populations in the muscle cells of desmin-deficient mice. The results conform to the conclusion that the reason for observed high apparent Km for ADP in regulation of oxidative phosphorylation in heart and slow twitch skeletal muscle cells in vivo is low permeability of mitochondrial outer membrane porins but not diffusion problems of ADP into and inside the cells. Most probably, in these cells there is a protein associated with cytoskeleton, which controls the permeability of the outer mitochondrial porin pores (VDAC) for ADP. Desmin itself does not display this type of control of mitochondrial porin pores, but its absence results in appearance of cells with disorganised structure and of altered mitochondrial population probably lacking this unknown VDAC controlling protein. Thus, there may be functional connection between mitochondria, cellular structural organisation and cytoskeleton in the cells in vivo due to the existence of still unidentified protein factor(s). PMID- 9398079 TI - Modification of the photosystem II acceptor side function in a D1 mutant (arginine-269-glycine) of Chlamydomonas reinhardti. AB - Bicarbonate anions have a strong positive influence on the electron and proton transfers in photosystem II (PS II). It has been suggested that bicarbonate binds to the non-heme iron and the QB binding niche of the PS II reaction center. To investigate the potential amino acid binding environment of bicarbonate, an arginine residue (R269) of the D1 protein of PS II of Chlamydomonas reinhardtii was mutated into a glycine; our characterization of the resultant mutant (D1 R269G) shows that both the TyrD+ and QA- Fe2+ EPR signals are substantially reduced and assembly of the tetranuclear Mn is lost (R.S. Hutchison, J. Xiong, R.T. Sayre, Govindjee, Biochim. Biophys. Acta 1277 (1996) 83-92). In order to understand the molecular implications of this mutation on the electron acceptor side of PS II, we used chlorophyll (Chl) a fluorescence as a probe of PS II structure and function, and herbicide binding as a probe for changes in the QB binding niche of PS II. Chl fluorescence measurements with the heterotrophically grown D1-R269G mutant cells (or thylakoids), as compared to that of the wild type, show that: rate of electron transfer from QA to the plastoquinone pool, measured by flash-induced Chl a fluorescence decay kinetics, is reduced by - 17 fold; the minimum Chl a fluorescence yield when all QA- is oxidized, is elevated by 2 fold; the level of stable charge separation as inferred from variable Chl fluorescence is reduced by 44%; binary oscillation pattern of variable Chl a fluorescence obtained after a series of light flashes is absent, indicative of the loss of functioning of the two-electron gate on the PS II acceptor side; 77 K PS II Chl a fluorescence emission bands (F685 and F695) are reduced by 20-30% (assuming no change in the PS I emission band). Thermoluminescence data with thylakoids show the absence of the S2QA- and S2QB- bands in the mutant. Herbicide 14C-terbutryn binding measurements, also with thylakoids, show that the QB niche of the mutant is significantly modified, at least 7-8 fold increased terbutryn dissociation constant is shown (220 nM in the mutant versus 29 nM in the wild type); the PS II sensitivity to bicarbonate-reversible formate inhibition is reduced by 5 fold in the mutant, although the formate/bicarbonate binding site still exists in the mutant. This suggests that D1-R269 must play some role in the binding niche of bicarbonate. On the basis of the above observations, we conclude that the D1-R269G mutation has not only altered the structure and function of PS II (QB niche being abnormal), but may also have a decreased net excitation energy transfer from the PS II core to the reaction center and/or an increased number of inactivated reaction center II. We also discuss a possible scenario for these effects using a recently constructed three dimensional model of the PS II reaction center. PMID- 9398080 TI - The subcellular sites of sphingomyelin synthesis in BHK cells. AB - The subcellular distributions of the enzymes which synthesise sphingomyelin (SM) and glucosylceramide (GluCer) from ceramide have been assessed in BHK cells. On a sucrose density gradient GluCer synthase (a marker of the cis/medial Golgi apparatus) and the trans-Golgi marker galactosyltransferase showed an similar monotonic distribution. In contrast, SM synthase showed two peaks of activity, a minor one which migrated with the Golgi markers and a major one which had a density close to that of plasma membrane markers (sphingomyelin, cholesterol, PtdSer, ganglioside GM3 and alkaline phosphodiesterase). When cell homogenates were treated with digitonin, the sedimentation characteristics of the Golgi markers was largely unaffected whereas the plasma membrane markers and the main peak of SM synthase activity were shifted to higher density. In contrast, when cells were treated with brefeldin A (BFA) the Golgi markers were shifted to higher density but not the plasma membrane markers or the main peak of SM synthase. These results suggest that the bulk of SM synthase activity in BHK cells is not associated with the Golgi cisternae but with a cell compartment which is relatively rich in cholesterol (e.g., plasma membrane, endosomes or trans-Golgi network.) Further experiments in which cells were treated with sphingomyelinase provided evidence that SM synthase activity was in an internal compartment and not at the plasma membrane. PMID- 9398081 TI - Lovastatin induces apoptosis by inhibiting mitotic and post-mitotic events in cultured mesangial cells. AB - Lovastatin, an inhibitor of protein prenylation, was reported to inhibit DNA synthesis and induce apoptosis in cultured cells. This report describes the morphological consequences of lovastatin treatment. Lovastatin (50 microM) induced mesangial cell rounding and disassembly of actin stress fibers within 24 to 48 h. After 48 to 72 h of lovastatin treatment, the cells detached from the substratum and underwent apoptotic cell death as evidenced by condensed nuclear chromatin, nuclear fragmentation, cell blebbing and decrease in cell size. Time lapse cinematography revealed that lovastatin caused cell rounding by either inhibiting cytokinesis or cell spreading following cytokinesis. Lovastatin induced cell rounding, detachment, and apoptosis were dependent upon cell proliferation. These effects were prevented by serum deprivation to inhibit cell proliferation or by plating cells at densities which resulted in contact inhibition of cell growth. Lovastatin-induced mesangial cell rounding and apoptosis were also prevented by the inclusion of the isoprenoids all-trans farnesol or all-trans-geranylgeraniol in the incubation medium. These results indicate that the effects of lovastatin were mediated by inhibition of protein isoprenylation because exogenous all-trans-geranylgeraniol can be used only in protein prenylation. The small GTP-binding protein RhoA, which may be important for cell spreading and cytokinesis, accumulated in the cytosol following treatment with lovastatin, suggestive of its inactivation. This effect was also prevented by the inclusion of either farnesol or geranylgeraniol in the incubation medium. Thus, lovastatin-induced apoptosis in mesangial cells occurs by interfering with prenylation dependent mitotic and post-mitotic events. PMID- 9398082 TI - Replenishment of selenium deficient rats with selenium results in redistribution of the selenocysteine tRNA population in a tissue specific manner. AB - We reported previously that the selenium status of rats influences both the steady-state levels and distributions of two selenocysteine tRNA isoacceptors and that these isoacceptors differ by a single methyl group attached to the ribosyl moiety at position 34. In this study, we demonstrate that repletion of selenium deficient rats results in a gradual, tissue-dependent shift in the distribution of these isoacceptors. Rats fed a selenium-deficient diet possess a greater abundance of the species unmethylated on the ribosyl moiety at position 34 compared to the form methylated at this position. A redistribution of the Sec tRNA isoacceptors occurred in tissues of selenium-supplemented rats whereby the unmethylated form gradually shifted toward the methylated form. This was true in each of four tissues examined, muscle, kidney, liver and heart, although the rate of redistribution was tissue-specific. Muscle manifested a predominance of two minor serine isoacceptors under conditions of extreme selenium-deficiency which also appeared to respond to selenium. Ribosomal binding studies revealed that one of the two additional isoacceptors decodes the serine codeword, AGU, and the second decodes the serine codeword, UCU. Interestingly, muscle and heart were the slower tissues to return to a 'selenium adequate' tRNA distribution pattern. PMID- 9398083 TI - Pepsinogen synthesis during long-term culture of porcine chief cells. AB - The purpose of this study was to characterize time-dependent changes in pepsinogen (PG) synthesis of porcine gastric chief cells during long-term monolayer culture. Porcine chief cells were isolated by pronase/collagenase treatment of fundic mucosa and enriched by density gradient and counterflow centrifugation. PG isoenzymes were identified in [L-35S]methionine-labelled cultured chief cells by native polyacrylamide gel electrophoresis followed by phosphor imager analysis, protease detection and immunoblots with specific PG A and C antibodies. The obtained results suggest that porcine chief cell cultures, after an initial settling period, reached an approximate steady state in total protein content and synthesis as well as in PG content and isoenzyme pattern from days 3 to 9 of culture. The latter was characterized by the presence of at least two PG A and two PG C isoenzymes. During the supposed steady-state total PG synthesis averaged out at 34 +/- 2% of total protein synthesis, as detected by [L 35S]methionine incorporation, due to the synthesis of, mainly, PG A2 and, to a much lesser extent, PG C and A1. In line with an active secretion, PG A2 proportion was on average significantly higher in released (44 +/- 3%) than in intracellular labelled proteins (19 +/- 2%). In addition, PG release from chief cells cultured for 6 and 9 days could be stimulated by cholecystokinin octapeptide. These data suggest that porcine chief cells in monolayer culture are a model well suited for the quantitative and qualitative characterization of PG isoenzyme synthesis and release during long-term investigations, for which an establishment of a culture steady state appears to be a useful prerequisite. PMID- 9398084 TI - Estrogen-stimulated transcytosis of desialylated ligands and alpha2 macroglobulin in rat liver. AB - Previous studies have shown that treatment of rats with 17 alpha-ethynylestradiol (EE) causes the appearance in bile of intravenously injected, desialylated ligands, including asialofetuin and low density lipoprotein (LDL). Here we show that activated alpha2-macroglobulin (alpha2-M*), but not insulin, transferrin or acetylated LDL, shows the same phenomenon. Alpha2-M* appearance in bile in EE treated rats was inhibited by receptor associated protein, but not unlabelled asialofetuin, strongly implicating the alpha2-macroglobulin receptor (alpha2MR/LRP) receptor in this process. Asialofetuin, apolipoprotein B (ApoB) of LDL and alpha2-M* appeared undegraded in the bile of EE-treated but not control rats. When LDL was injected, not only was intact apolipoprotein B detected in bile, but the profile of cholesterol esters appearing in bile was characteristic of the injected human LDL rather than rat lipoproteins. After floatation of the bile on KBr gradients, intact Apo B and cholesterol esters characteristic of human LDL were found at the normal density of LDL suggesting that the majority of the lipoprotein particle remains intact. Stimulation of transcytosis was specific to estrogens, and was highest with 17alpha-ethynylestradiol. After subcutaneous injection of 0.05 mg/kg body weight of ethynylestradiol, sufficient to give a measurable increase in transcytosis, the plasma concentration of ethynylestradiol rose to 2.2 nM. Thus estrogen-stimulated transcytosis of desialylated ligands and alpha2-M* would be expected at physiological estrogen concentrations. PMID- 9398085 TI - Activation of inducible nitric oxide synthase by human choriogonadotropin in RAW 264.7 cells. AB - Although human choriogonadotropin (hCG) plays a crucial role in the regulation of the menstrual cycle and maintenance of pregnancy, little is known about the other functions. However, recently hCG receptors have been identified in nongonadal cells. The objective of the current study was to determine the effect of hCG on the production of nitric oxide (NO). Stimulation of RAW 264.7 cells with hCG after treatment with recombinant interferon-gamma (rIFNgamma) resulted in increased NO synthesis. hCG had no effect on NO synthesis itself. NO production was inhibited by N(G)-monomethyl-L-arginine. rIFNgamma in combination with hCG showed marked increase of the expression of the inducible nitric oxide synthase (iNOS) gene. In addition, synergy between rIFNgamma and hCG was mainly dependent on hCG-induced tumor necrosis factor-alpha (TNF-alpha) secretion. All the preparations of hCG were endotoxin free. These results suggest that the capacity of hCG to increase NO production from rIFNgamma-primed RAW 264.7 cells is the result of hCG-induced TNF-alpha secretion. PMID- 9398086 TI - Induction of thioredoxin in human lymphocytes with low-dose ionizing radiation. AB - Induction of the expression of the thioredoxin (TRX) gene, producing a key protein in regulating cellular functions through redox reaction as well as being a radioprotector, was followed after ionizing irradiation of lymphocytes from human donors. The TRX mRNA level increased to a peak, 5.7-fold higher than the control at maximum, 6 h after irradiation, and then decreased. The optimum radiation dose for enhancement of induction of the TRX mRNA was 0.25 Gy. The TRX protein also increased to a peak, a 3-fold increase at maximum, with the same timing as that for TRX mRNA. PMID- 9398087 TI - Differanisole A, a novel antitumor antibiotic, enhances growth inhibition and differentiation of human myeloid leukemia cells induced by 9-cis retinoic acid. AB - Differanisole A, 3,5-dichloro-2-hydroxy-4-methoxy-6-n-propylbenzoic acid, inhibited growth of human myeloid leukemia cells. The compound induced G1 arrest and granulocytic differentiation of HL-60 cells, although the differentiation inducing effect was modest. Differanisole A and 9-cis retinoic acid (9cisRA) synergistically inhibited the growth and induced functional and morphologic differentiation of HL-60 and NB4 cells, whereas the combined treatment with differanisole A and all-trans retinoic acid or 1alpha,25-dihydroxyvitamin D3 was less effective. Similar results were obtained in primary culture of leukemia cells from a patient with acute promyelocytic leukemia. The synergistic effect on growth inhibition and induction of differentiation required simultaneous treatment with differanisole A and 9cisRA. Differanisole A and an RXR-specific ligand (Ro47-5944) cooperatively inhibited the cell growth, while the combined effect of differanisole A and an RAR-specific ligand Am80 was just additive. Differanisole A in combination with 9cisRA may have implications for therapy of acute promyelocytic leukemia patients. PMID- 9398088 TI - Leukotriene B4 stimulates the release of arachidonate in human neutrophils via the action of cytosolic phospholipase A2. AB - Leukotriene B4 (LTB4) is a potent lipid mediator of inflammation and is involved in the receptor-mediated activation of a number of leukocyte responses including degranulation, superoxide formation, and chemotaxis. In the present research, stimulation of unprimed polymorphonuclear leukocytes (neutrophils) with LTB4 results in the transient release of arachidonate as measured by mass. This release of arachidonate was maximal at an LTB4 concentration of 50-75 nM and peaked at 45 s after stimulation with LTB4. The transient nature of this release can be attributed, in part, to a fast (< 60 s) metabolism of the added LTB4. Moreover, the inhibition of the reacylation of the released arachidonate with thimerosal results in greater than 4-times as much arachidonate released. Thus, a rapid reacylation of the released arachidonate also contributes to the transient nature of its measured release. Multiple additions of LTB4, which would be expected to more closely resemble the situation in vivo where the cell may come into contact with an environment where LTB4 is in near constant supply, yielded a more sustained release of arachidonate. No release of [3H]arachidonate was observed when using [3H]arachidonate-labeled cells. This indicates that the release of arachidonate as measured by mass is most probably the result of hydrolysis of arachidonate-containing phosphatidylethanolamine within the cell since the radiolabeled arachidonate is almost exclusively incorporated into phosphatidylcholine and phosphatidylinositol pools under the non-equilibrium radiolabeling conditions used. Consistent with the role of cytosolic phospholipase A2 (cPLA2) in the release of arachidonate, potent inhibition of the LTB4-stimulated release was observed with methylarachidonylfluorophosphonate, an inhibitor of cPLA2 (IC50 of 1 microM). The bromoenol lactone of the calcium independent phosphospholipase A2. failed to affect LTB4-stimulated release of arachidonate in these cells. PMID- 9398089 TI - Expression of calmodulin and calmodulin binding proteins in rat fibroblasts stably transfected with protein kinase C and oncogenes. AB - Molecular mechanisms leading to elevated calmodulin (CaM) expression in cancer have not yet been discovered. We have quantitated the levels of transcripts derived from all three CaM genes in a variety of the same origin rat fibroblasts transformed with oncogenes in combination with gene for protein kinase C using Northern blot analysis with three CaM gene specific cDNA probes. Five species of CaM mRNA were detected in all these cells. Surprisingly many of the investigated cell lines exhibited a decreased content of all CaM mRNAs as compared to control cells with CaMI and CaMII transcripts showing the most pronounced alterations. In contrast, CaM protein levels were increased in all these cell lines as determined by a radioimmunoassay. These results suggest that oncogenic up-regulation of CaM synthesis takes place posttranscriptionally. Several CaM binding proteins were found at different concentrations in the studied cell lines depending on the oncogenes used for transformation. However, CaM overexpression does not seem to affect the overall levels of CaM binding proteins. PMID- 9398090 TI - Baseline characteristics of patients in the coronary artery bypass graft (CABG) Patch Trial. AB - BACKGROUND: Patients with left ventricular dysfunction who undergo coronary artery bypass graft (CABG) surgery frequently have late sudden cardiac death. The CABG Patch Trial is a prospective, randomized, multicenter clinical trial that randomized patients at high risk at the completion of CABG surgery to implantation of an epicardial implantable cardioverter defibrillator (ICD) or to no antiarrhythmic treatment. The trial was designed to determine whether prophylactic implantation of an ICD at the time of CABG surgery would result in a lower total mortality in long-term follow-up. METHODS: Patients undergoing CABG surgery were eligible for the trial if they were younger than 80 years, had a left ventricular ejection fraction less than 0.36, and had an abnormal signal averaged electrocardiogram. Patients with a history of sustained ventricular tachycardia or ventricular fibrillation were excluded from the trial. All patients were scheduled to undergo follow-up at 3-month intervals until 42 months after surgery. RESULTS: Randomization of patients in the trial ended in February 1996. During the recruitment period 71,855 patients were screened, 1,422 were eligible, 1,055 were enrolled (signed consent forms), and 900 patients (76% of eligible patients) were randomized. The mean age of the 446 patients in the ICD group was 64 years versus 63 years for the 454 patients in the control group. A total of 87% of the participants in the ICD group were men versus 82% in the control group (p = NS). Most of the patients had a history of hypertension (55%), smoking (78%), and hypercholesterolemia (54%). Half of the patients had clinical heart failure, and the mean ejection fraction for both patient groups was 0.27 +/ 0.06. No difference was seen in the history of myocardial infarction (83%), congestive heart failure (50%), or atrial (11%) or ventricular (17%) arrhythmias between the two groups. Major clinical characteristics (age, sex, number of previous infarctions, incidence of heart failure, and mean left ventricular ejection fraction) were almost identical to those found in another ICD primary prevention trial, the Multicenter Automatic Defibrillator Implantation Trial (MADIT). CONCLUSIONS: A high risk sample of patients was enrolled in The CABG Patch Trial, as shown by examination of their baseline characteristics. PMID- 9398091 TI - Differential effects of adenosine on antegrade and retrograde fast pathway conduction in atrioventricular nodal reentry. AB - Although adenosine depresses antegrade atrioventricular (AV) nodal conduction, the effects of adenosine on antegrade and retrograde fast pathway conduction in AV nodal reentry have not been determined. In 17 patients (five men, 12 women, mean age 49 +/- 12 years) with common slow-fast AV nodal reentrant tachycardia, the antegrade slow pathway conduction was selectively and completely ablated by radiofrequency catheter ablation while the antegrade and retrograde fast pathway conduction remained intact. During high right atrial pacing at a mean pacing cycle length of 474 +/- 36 msec, adenosine was rapidly injected intravenously at an initial dose of 0.5 mg followed by stepwise increases of 0.5 mg or 1.0 mg given at 5-minute intervals until second-degree AV block developed. During right ventricular apical pacing at the same pacing cycle lengths (mean 474 +/- 36 msec) as those in the study of antegrade conduction, intravenous injection of incremental doses of adenosine was repeated until ventriculoatrial (VA) block occurred. The adenosine-induced prolongation of VA conduction was also determined in the presence of verapamil (loading dose 0.15 mg/kg, maintenance dose 0.005 mg/kg/min) in seven of 17 patients. The dose of adenosine required to produce AV block, the increase in the atrio-His interval by 50% and the maximal response were 3.4 +/- 1.4 mg, 1.8 +/- 0.6 mg, and 58% +/- 5%, respectively. On the other hand, the dose of adenosine required to produce VA block, the increase in the VA interval by 50%, and the maximal response were 8.2 +/- 2.9 mg, 3.4 +/- 0.6 mg, and 20% +/- 5%, respectively, in the control and 3.7 +/- 0.5 mg, 3.5 +/- 0.7 mg, and 23% +/- 5%, respectively, in the presence of verapamil. In conclusion, adenosine has a differential potency to depress AV and VA conduction in patients with AV nodal reentry, with greater potency for slowing antegrade fast than retrograde fast pathway conduction. Verapamil had an additive effect to adenosine on slowing retrograde VA conduction, which further supports the evidence that the retrograde fast pathway in part involves an AV nodal-like structure. PMID- 9398092 TI - Sex and diagnostic evaluation of possible coronary artery disease after exercise treadmill testing at one academic teaching center. AB - Controversy exists as to whether a sex bias exists that affects the diagnostic approach to suspected coronary artery disease: previous studies have used coronary angiography, but not other noninvasive testing, as a primary end point. This investigation examined posttest sex differences in diagnostic evaluation after exercise treadmill testing according to a broader end point than just coronary angiography alone. The design was a cohort analytic study with a 90-day follow-up. The study was done at the Cleveland Clinic Foundation, an academic group practice. Patients included consecutive adults (1023 men and 579 women) with chest pain but no documented coronary disease who were referred for symptom limited exercise treadmill testing without adjunctive imaging; none had undergone prior invasive cardiac procedures. Main outcome measures included (1) performance of any subsequent diagnostic study (invasive or noninvasive) and (2) performance of coronary angiography as the next diagnostic study. During follow-up, 89 (8.7%) men and 48 (8.3%) women underwent a second diagnostic study (odds ratio 0.95; 95% confidence interval 0.66 to 1.37; p > 0.8), whereas 64 (6.3%) men and 21 (3.6%) women went straight to coronary angiography (odds ratio 0.56; 95% confidence interval 0.34 to 0.93; p = 0.02). In multivariable logistic regression analyses, which considered baseline clinical characteristics, the ST-segment response, and other prognostically important exercise responses, women tended to be less likely than men to be referred to any second test (adjusted odds ratio 0.70; 95% confidence interval 0.42 to 1.19; p > 0.1) but were markedly and significantly less likely to be referred straight to coronary angiography (adjusted odds ratio 0.33; 95% confidence interval 0.17 to 0.65). After exercise treadmill testing, women were only slightly less likely than men to be referred for subsequent diagnostic testing; they were, however, much less likely to be referred straight to coronary angiography as opposed to another noninvasive study. PMID- 9398093 TI - Two-dimensional echocardiographic assessment of the progression of aortic root size in 127 patients with chronic aortic regurgitation: role of the supraaortic ridge and relation to the progression of the lesion. AB - Although aortic root dilation has etiologic and prognostic significance in patients with chronic aortic regurgitation (AR), no information is available regarding changes over time in aortic root size in patients with the entire spectrum of AR severity or how such changes relate to progression of the AR or to left ventricular (LV) overload. To analyze this, a total of 127 patients with chronic AR who had more than 6 months of follow-up by two-dimensional and Doppler echocardiography were included in the study (69 men and 58 women; mean age 59.3 +/- 21.2 years [range 14 to 94 years]; 67 cases of mild, 45 moderate, 15 severe, and 21 bicuspid aortic valve disease). The aortic anulus, sinuses of Valsalva, supraaortic ridge, and ascending aorta were measured in the parasternal long-axis view, LV volumes were calculated (biplane Simpson's approach), and the severity of AR was quantified based on proximal jet size and graded according to an algorithm that takes into account major color Doppler criteria. At entry to the study, significant differences between patients with mild, moderate, and severe AR were noted only in supraaortic ridge size (1.46 +/- 0.29 cm/m2 vs 1.63 +/- 0.33 cm/m2 [p < 0.006]; vs 1.67 +/- 0.43 cm/m2 [p < 0.03]). A significant increase in aortic root size at all levels was observed during the follow-up period in all three groups of severity of AR. The rate of change of the supraaortic ridge, the upper support structure of the anulus and cusps, was faster in patients with more severe degrees of AR (p = 0.013); this was not the case at the other aortic levels. No differences were observed in aortic root size or rate of progression between patients with bicuspid or tricuspid aortic valves. Patients were considered "progressive" if they lay on the steepest positive segment of the curve representing the rank order in the rate of aortic root progression. Compared with "nonprogressive" patients, patients who were progressive in supraaortic ridge size (rate >0.12 cm/yr; n = 23) had a faster rate of progression in the degree of regurgitation as assessed by the regurgitant jet area/LV outflow tract area ratio measured in the parasternal short-axis view (0.48 +/- 0.45 vs 0.24 +/- 0.5/yr; p < 0.03) and a foster rate of progression of LV end-diastolic volume (30 +/- 22.8 vs 14.4 +/- 15.6 ml/yr; p < 0.0002) and LV mass (70.8 +/- 74.4 vs 16.8 +/- 19.2 gm/yr; p < 0.0004). In conclusion, there is progressive dilation of the aortic root at all levels, even in patients with mild AR. More rapid progression in aortic root size is associated with more rapid progression of the underlying aortic insufficiency, as well as more rapid increases in LV volume and mass. PMID- 9398094 TI - Low dose dobutamine echocardiography is more predictive of reversible dysfunction after acute myocardial infarction than resting single photon emission computed tomographic thallium-201 scintigraphy. AB - To directly compare dobutamine echocardiography and resting single photon emission computed tomographic (SPECT) thallium-201 (Tl-201) scintigraphy for the detection of reversible dysfunction, 64 patients underwent dobutomine echocardiography (baseline, low dose 5 and 10 mg/kg/min, and peak dose), rest Tl 201 scintigraphy (3 mCi - 15 minute and 3- to 4-hour SPECT imaging), and coronary angiography during the first week after acute myocardial infarction. Follow-up echocardiography was performed 4 to 8 weeks after discharge. Wall thickening improved at follow-up in 52% (207 of 399) of the dysfunctional segments. By receiver operating characteristic analysis, biphasic responses and sustained improvement during dobutamine echocardiography were more accurate (p < 0.01) than Tl-201 uptake by SPECT scintigraphy for reversible dysfunction. The greater accuracy of dobutamine echocardiography resulted from higher accuracy in akinetic segments, Q wave infarction, and multivessel coronary artery disease. In conclusion, dobutamine echocardiography was more accurate than resting SPECT Tl 201 scintigraphy for reversible dysfunction after acute myocardial infarction. PMID- 9398095 TI - Myocardial viability in patients with chronic coronary artery disease and previous myocardial infarction: comparison of myocardial contrast echocardiography and myocardial perfusion scintigraphy. AB - The aim of this study was to compare perfusion patterns on myocardial contrast echocardiography with those on myocardial perfusion scintigraphy for the assessment of myocardial viability in patients with previous myocardial infarction. Accordingly, perfusion scores with the two techniques were compared in 91 ventricular regions in 21 patients with previous (>6 weeks old) myocardial infarction. Complete concordance between the two techniques was found in 63 (69%) regions; 25 (27%) regions were discordant by only 1 grade, and complete discordance (2 grades) was found in only 3 (3%) regions. A kappa statistic of 0.65 indicated good concordance between the two techniques. Although the scores on both techniques demonstrated a relation with the wall motion score, the correlation between the myocardial contrast echocardiography and wall motion scores was closer (r = -0.63 vs r = -0.50, p = 0.05). It is concluded that myocardial contrast echocardiography provides similar information regarding myocardial viability as myocardial perfusion scintigraphy in patients with coronary artery disease and previous myocardial infarction. PMID- 9398096 TI - Dichloroacetate as metabolic therapy for myocardial ischemia and failure. AB - This article critically reviews the pharmacologic effects of the investigational drug dichloroacetate (DCA), which activates the mitochondrial pyruvate dehydrogenase enzyme complex in cardiac tissue and thus preferentially facilitates aerobic oxidation of carbohydrate over fatty acids. The pharmacologic effects of DCA are compared with other interventions, such as glucose plus insulin, inhibitors of long chain fatty acid oxidation and adenosine, that are also thought to exert their therapeutic effects by altering myocardial energy metabolism. Short-term clinical and laboratory experiments demonstrate that intravenous DCA rapidly stimulates pyruvate dehydrogenase enzyme complex activity and, therefore, aerobic glucose oxidation in myocardial cells. Typically these effects are associated with suppression of myocardial long chain fatty acid metabolism and increased left ventricular stroke work and cardiac output without changes in coronary blood flow or myocardial oxygen consumption. Although long term studies are lacking, short-term parenteral administration of DCA appears to be safe and capable of significantly improving myocardial function in conditions of limited oxygen availability by increasing the efficient conversion of myocardial substrate fuels into energy. PMID- 9398097 TI - Coronary artery bypass grafting in the elderly. PMID- 9398098 TI - Risk factors for life-threatening cavopulmonary thrombosis in patients undergoing bidirectional superior cavopulmonary shunt: an exploratory study. AB - We have observed six patients with life-threatening superior vena caval or pulmonary thrombosis after bidirectional superior cavopulmonary shunt. With the use of a case control study we sought to identify perioperative risk factors for this thrombotic complication. Medical records of six patients with cavopulmonary thrombosis and those of 24 patients in a control group were reviewed to abstract data for potential risk factors. Contingency tables and univariate logistic regression were used to determine associations between various perioperative parameters and occurrence of cavopulmonary thrombosis. Preoperative variables associated with thrombosis included bilateral superior vena cavae, odds ratio: 23, p = 0.02, increased age at surgery (p = 0.05), and female sex (odds ratio: 7, p = 0.05). The McGoon Ratio (index of relative pulmonary artery branch diameter) was inversely related to thrombosis risk (p = 0.08). Two torr increases in mean right atrial (p = 0.08) or ventricular end-diastolic (p = 0.05) pressures were associated with approximately 70% increases in thrombosis risk. Intraoperative prolongation of aortic cross-clamp time related directly to thrombosis risk (p = 0.06). Postoperative variables associated with thrombosis included increased superior vena caval pressure within 12 hours after surgery (odds ratio > or = 10 for 5 torr increase in pressure, p = 0.02) and poor ventricular function (odds ratio: 9, p = 0.06) We conclude that high risk variables for patients undergoing a cavopulmonary shunt include bilateral superior vena cavae, female sex, increasing age, decreased McGoon Ratio, and elevated right atrial and ventricular end-diastolic pressure (before surgery), patients with prolonged aortic cross clamp time (during surgery), and patients with elevated superior vena caval pressure and poor ventricular function (after surgery). PMID- 9398099 TI - Effects of amlodipine in patients with chronic heart failure. AB - The role of calcium antagonists in patients with ischemic heart failure is currently unclear. We examined the effects of amlodipine on exercise capacity and central and regional hemodynamics in 32 patients with mild to moderate chronic heart failure in a single-center, double-blind, randomized placebo-controlled trial. All were taking at least 40 mg of furosemide daily with an angiotensin converting enzyme inhibitor. Ischemic heart disease was the most common cause of heart failure, but no patient had symptom-limiting angina. Mean treadmill exercise capacity in patients taking amlodipine increased by 96 seconds (95% confidence interval -23 to 215) and 50 seconds (-34 to 135) in the placebo group; mean difference in change between treatments was 70 seconds (-90 to 233), p = 0.38. Active treatment with amlodipine did not affect self-paced corridor walking times. Similarly, there were no significant effects on cardiac output, oxygen uptake, heart rate, and mean arterial pressure at rest or during exercise. Calf and renal blood flow were also unchanged by treatment. The lack of significant effect demonstrated by these data suggests a limited role for amlodipine in patients with ischemic cardiomyopathy, although it may prove beneficial in those with nonischemic disease. More data are required before amlodipine can be recommended for all patients with chronic heart failure. PMID- 9398100 TI - Depressed arterial baroreflex sensitivity and not reduced heart rate variability identifies patients with chronic heart failure and nonsustained ventricular tachycardia: the effect of high ventricular filling pressure. AB - In chronic heart failure (CHF) the contributing role of increased sympathetic activity and hemodynamic dysfunction in the genesis of ventricular arrhythmias has not been well established. To assess the relation between severe ventricular arrhythmias, hemodynamic impairment, and autonomic nervous system derangement, 142 patients with CHF in sinus rhythm underwent 24-hour electrocardiographic recording, right-sided heart catheterization, and evaluation of sympathovagal balance by heart rate variability (HRV) and baroreflex sensitivity (BRS). Patients were grouped according to the absence (without nonsustained ventricular tachycardia [NSVT]; n = 87) or presence (with NSVT; n = 55) of NSVT. Patients with NSVT had higher pulmonary artery and capillary pressures and more pronounced signs of sympathetic activation and parasympathetic withdrawal compared with those without NSVT. However, logistic regression analysis revealed that depressed BRS but not reduced HRV was significantly associated with the presence of NSVT, at both univariate analysis and after adjustment for clinical and hemodynamic variables. Moreover, it was found that when depressed BRS was associated with high pulmonary capillary pressure, the odds ratio for having NSVT rose markedly from 3.8 to 6.5. In conclusion, this study indicates that in stable CHF the assessment of arterial baroreflex function, but not HRV analysis, allows identification of patients at high risk of NSVT. It is suggested that the effect of depressed BRS is strengthened by the simultaneous presence of increased myocardial wall stress. These data support the hypothesis of a contributory role of autonomic nervous system dysfunction as expressed by the inability to activate effective vagal reflexes and an indirect index of ventricular stretch in the genesis of life-threatening arrhythmias. PMID- 9398101 TI - Continuous home ambulatory intravenous inotropic drug therapy in severe heart failure: safety and cost efficacy. AB - Some patients with dilated cardiomyopathy who are inotrope dependent but remain well by undergoing infusions can be managed by ambulatory infusions at home. We report our results in 20 patients awaiting heart transplantation, unable to be weaned from intravenous inotropic therapy on 2 or more occasions, but who were well while receiving inotropes and received home ambulatory infusions. The patients were treated with ACE inhibitors, digoxin, diuretics, vasodilators, close electrolyte management, and low-dose amiodarone for those with more than four-beat ventricular tachycardia. Infusions were delivered by a tunneled subclavian catheter and syringe driver. Thirteen patients received dopamine, four received dobutamine, and three received both. Mean duration of inotropic therapy was 5 months with 70% of the time spent as an outpatient. Eleven patients received transplants, two remain on the waiting list, and seven died after being removed from the list because of general deterioration or renal dysfunction. There were no sudden deaths. Actuarial survival was 71% at 3 months, which is not less than that expected for an inotrope-dependent population. All patients with idiopathic dilated cardiomyopathy survived to transplantation. In contrast, all three with right heart failure caused by pulmonary vascular disease and four of seven with ischemic cardiomyopathy died. Inpatient days were reduced by 70%, leading to considerable cost savings. Home ambulatory inotropic therapy is safe, cost-effective, best suited to those with idiopathic dilated cardiomyopathy, and dramatically reduces inpatient hospital duration. PMID- 9398102 TI - Heart failure between 1986 and 1994: temporal trends in drug-prescribing practices, hospital readmissions, and survival at an academic medical center. AB - Since 1987, publications in widely circulated medical journals have reported improved survival and lower hospital readmission rates when patients with heart failure and systolic dysfunction are treated with angiotensin-converting enzyme (ACE) inhibitors. We describe changes in ACE inhibitor use among patients hospitalized with heart failure between 1986 and 1993. Simultaneous trends in readmissions and survival rates are reported. Subjects were 612 consecutive patients hospitalized with a principal diagnosis of heart failure at an academic medical center during the period of Sept. 1, 1986, to Dec. 31, 1987 (interval I) or during the period Aug. 1, 1992, to Nov. 30, 1993 (interval II). Medical records were reviewed for 434 patients, consisting of all patients hospitalized with heart failure during interval II and a randomly selected 50% subset of patients hospitalized during interval I. Among 145 patients with systolic dysfunction whose medical records were reviewed, ACE inhibitor prescriptions significantly increased between interval I and interval II (43% vs 71%, p < 0.01, odds ratio 3.22, 95% confidence interval 1.62 to 6.42). Prescriptions of ACE inhibitors combined with digoxin and a diuretic also increased (37% vs 56%, p = 0.02, odds ratio 2.22, 95% confidence interval 1.14 to 4.32). Among all 612 patients, 6-month heart failure readmission rates increased from 13% to 21% (p = 0.02, odds ratio 1.79, 95% confidence interval 1.10 to 2.82). There was no significant change in survival rate between interval I and interval II, however, survival rate was marginally significantly improved among patients with systolic dysfunction. Our results suggest that drug-prescribing practices have significantly changed between 1986 and 1993. The absence of observed improvement in outcomes may result from changes in hospital admission criteria for heart failure. PMID- 9398103 TI - Digitalis increases brain natriuretic peptide in patients with severe congestive heart failure. AB - Ouabain can cause increased secretion of atrial natriuretic peptide (ANP) from atrial cardiocyte culture, but the effects of digitalis in a therapeutic range on the secretion of cardiac natriuretic peptide including ANP and brain natriuretic peptide (BNP), mainly from the ventricle, in patients with congestive heart failure remain to be investigated. Therefore we studied the acute effects of intravenous infusion of a relatively low dose of digitalis or placebo on hemodynamics and neurohumoral factors including the plasma levels of ANP and BNP and cyclic guanosine monophosphate, a second messenger of cardiac natriuretic peptide, in 13 patients with severe congestive heart failure. No significant change in the hemodynamic parameters or neurohumoral factors was observed with placebo. After 1 hour of intravenous administration of deslanoside (0.01 mg/kg), there was a significant decrease of plasma renin activity and angiotensin II, aldosterone, and norepinephrine levels but no significant change of plasma levels of vasopressin and a significant decrease of the pulmonary capillary wedge pressure but no significant change in cardiac index. In addition, plasma levels of ANP (217 +/- 47 vs 281 +/- 70 pg/ml, p < 0.05), BNP (628 +/- 116 vs 689 +/- 132 pg/ml, p < 0.05), and cyclic guanosine monophosphate (9.7 +/- 1.1 vs 10.9 +/- 1.5 pmol/ml, p < 0.05) increased despite the decrease of pulmonary capillary wedge pressure (19.7 +/- 2.3 vs 16.8 +/- 2.3 mm Hg, p < 0.05). These results indicate the acute intravenous low dose of digitalis resulted in a significant increase in plasma levels of ANP, BNP, and cyclic guanosine monophosphate concomitant with the significant decrease of pulmonary capillary wedge pressure, suggesting the acute direct action of digitalis on the cardiac natriuretic peptides released from the heart in patients with severe congestive heart failure. PMID- 9398104 TI - Balloon angioplasty of native aortic coarctation in infants 3 months of age and younger. AB - The use of balloon dilation to treat native aortic coarctation is controversial, particularly in infants. Between January 1991 and September 1996, 12 patients < or = 3 months of age with native coarctation of the aorta (CoA) underwent balloon angioplasty (BA). All 12 lesions were dilated successfully with a mean reduction in peak systolic gradient from 49.3 +/- 16.5 mm Hg to 6.8 +/- 4.0 mm Hg (p < 0.001) and a mean increase in minimum CoA diameter from 2.4 +/- 0.6 mm to 5.5 +/- 1.3 mm (p < 0.001). Intimal flaps or tears were detected immediately after BA in 4 (33%) of 12 patients by angiography and in 8 (89%) of 9 patients by intravascular ultrasonography. No deaths or major complications related to the BA occurred. One patient had documented asymptomatic femoral artery obstruction, and one patient with hydrops fetalis and congenital pleural effusions died with gram negative sepsis 1 week after the procedure. Follow-up was available for 10 patients (1 was lost to follow-up) between 2 months and 4.1 years (mean 2.4 +/- 1.3 years) after BA. No patient had an aortic aneurysm. Restenosis occurred in 5 (50%) of 10 patients, requiring reintervention a mean of 2.6 +/- 2.1 months after BA. One patient underwent surgical repair. Repeat BAs were performed in the other four patients; three were successful, and one with partial gradient relief required surgical repair. Five patients have not required reintervention a mean of 2.9 +/- 1.0 years after the initial BA. Among these five patients, follow-up intravascular ultrasound performed in three patients a mean of 2.0 +/- 1.9 years after BA showed favorable endovascular remodeling. There was a tendency for early reintervention in patients < 1 month of age and coexistence of a patent ductus arteriosus at the time of BA. In conclusion, selected infants < or = 3 months of age with discrete native CoA may be treated initially with balloon dilation. Most patients who have restenosis respond successfully to repeat BA. PMID- 9398105 TI - Aortic valve resistance in aortic stenosis: Doppler echocardiographic study and surgical correlation. AB - Four hundred seven patients with aortic stenosis who had Doppler echocardiography before surgery were studied to determine the feasibility of Doppler-derived valve resistance calculation and its clinical value. Patients with milder aortic stenosis had lower mean gradient, larger valve area, and lower maximal resistance than those with severe stenosis. Maximal resistance was related strongly to aortic stenosis severity but did not add any information after valve area and gradient were known and was not related to surgical mortality. PMID- 9398106 TI - Coronary artery stenting in cardiac allograft vascular disease. AB - Cardiac allograft vascular disease is characterized by diffuse and multifocal heterogeneous myointimal hyperplasia with or without vascular remodeling. Catheter-based interventions are indicated in selected patients. This study documents our experience with percutaneous transluminal coronary angioplasty and coronary stents (n = 48) in a group of 27 patients 5.7 +/- 2.9 years after heart transplantation. Early and intermediate results were controlled by angiography and intravascular ultrasound. Conventional percutaneous transluminal coronary angioplasty resulted in a mild and mostly inadequate gain in luminal dimensions (lumen area: 3.17 +/- 0.92 mm2 to 3.70 +/- 1.21 mm2; minimal lumen diameter: 1.84 +/- 0.23 mm to 2.04 +/- 0.36 mm). Coronary stenting led to a further improvement of luminal gain (lumen area: 3.70 +/- 1.21 mm2 to 5.86 +/- 1.76 mm2; minimal lumen diameter: 2.04 +/- 0.36 mm to 2.53 +/- 0.38 mm). These results were stabilized by application of aspirin and ticlopidine only. There were no stent thromboses or bleeding complications, and early hospital discharge of the patients was possible. At follow-up (mean follow-up period 7.72 +/- 5.45 months (range 0.50 to 23.13 months) all patients were clinically event free. In six of 24 stented vessels (25%) in 16 patients, significant restenosis (>50%) was found by intravascular ultrasound (n = 20) or by angiography (n = 4) 6 months after stent placement. We conclude that in eligible cardiac allograft vascular disease lesions primary stenting may be the method of choice. However, further evaluation of the modalities of stent application and different stent designs with respect to long-term survival is necessary. PMID- 9398107 TI - Feasibility and applicability of coronary stent implantation with the direct brachial approach: results of a single-center study. AB - Implantation of stents in selected patients improves outcome after coronary angioplasty. Newer antiplatelet regimes limit access site complications associated with stenting by the percutaneous femoral approach, but a substantial proportion of patients will require anticoagulant therapy for concomitant disease or will have peripheral vascular disease that prevents access from the leg. We investigated procedural success rates and outcome in consecutive patients undergoing elective stent implantation in our institution. In 73 patients who were receiving anticoagulation therapy and were stented by a direct approach to the left brachial artery, 98.6% of stents were successfully deployed, with a major vascular access site complication rate of 1.4%. Equipment consumption, procedural success rate, and fluoroscopy time were similar in patients stented by the direct brachial or percutaneous femoral approach. Where the percutaneous femoral approach is precluded or patients are anticoagulated, stent procedures can be successfully performed by the direct brachial approach with a low rate of access site complications, even when large-caliber guiding catheters are required. PMID- 9398108 TI - Effect of carteolol on silent myocardial ischemia, variability in heart rate, and the pain-modulating system. AB - To investigate the effects of carteolol, which is a nonselective beta-adrenergic agent with intrinsic sympathomimetic activity, on silent myocardial ischemia, exercise-induced myocardial ischemia, indexes of heart rate variability, and pain modulating system, 20 patients (mean 60 +/- 9 years) with chronic stable angina underwent exercise treadmill testing and 24-hour ambulatory electrocardiographic monitoring during 2 weeks of carteolol administration (15 mg/day) in a double blind, placebo-controlled design. Plasma levels of beta-endorphin and bradykinin and electrical pain stimulation to the skin were measured at rest and peak exercise. Indexes of heart rate variability of both time-domain and frequency domain analysis were derived from 24-hour ambulatory electrocardiographic monitoring. Carteolol decreased maximal heart rate responses to daily activities during ambulatory monitoring and significantly reduced the median frequency and duration of silent myocardial ischemic episodes (from 1.0 to 0.0 events/24 hr and from 16 to 0 min/24 hr, respectively). Carteolol significantly decreased the rate pressure product at rest and during exercise with improving maximal ST segment depression, suggesting amelioration of exercise-induced myocardial ischemia. Carteolol did not significantly affect plasma levels of beta-endorphin and bradykinin or pain threshold. It significantly decreased some indexes (standard deviation of all normal sinus R-R intervals in the entire 24-hour recording and standard deviation of the mean of all 5-minute segments of normal R-R intervals of a 24-hour recording) of heart rate variability. These results suggest that carteolol may reduce total myocardial ischemic burden by the reduction of cardiac oxygen demand during daily activities and exercise stress, while not affecting plasma levels of beta-endorphin, bradykinin, and pain threshold. Because carteolol tended to decrease indexes of heart rate variability, significant caution might be necessary in prescribing the beta-blocking agents with intrinsic sympathomimetic activity like carteolol to patients with potential serious arrhythmia. PMID- 9398109 TI - Clinical implications of cigarette smoking in acute myocardial infarction: acute angiographic findings and long-term prognosis. AB - This study was undertaken to assess whether reperfusion in smokers could be achieved spontaneously or therapeutically and to assess whether favorable outcome in smokers could be sustained for years after infarction. We studied 260 patients with anterior myocardial infarction who underwent coronary angiography and thrombolysis within 24 hours after the onset of chest pain. There were 158 smokers and 102 nonsmokers. Smoking was associated more with men, younger age, and less multivessel disease. On initial angiography, the distribution of Thrombolysis in Myocardial Infarction grade was similar between smokers and nonsmokers. After thrombolysis, Thrombolysis in Myocardial Infarction grade 3 was more frequent in smokers (32% vs 18%; p = 0.004). In-hospital mortality rates were lower (8% vs 18%; p = 0.022) and long-term cardiac survival was better in smokers (5-year survival: 82% vs 70%; p = 0.022). Our data demonstrated that the infarct artery of smokers responded more efficiently to thrombolysis and favorable outcome in smokers was sustained throughout 5 years. PMID- 9398110 TI - The effects of the angiotensin-converting enzyme inhibitor imidapril on plasma plasminogen activator inhibitor activity in patients with acute myocardial infarction. AB - This study sought to determine whether early treatment with angiotensin converting enzyme (ACE) inhibitors in patients with acute myocardial infarction (AMI) is useful for the improvement of fibrinolytic function, as well as left ventricular function. This study was designed to examine the levels of plasma plasminogen activator inhibitor (PAI) activity and serum ACE activity during the course of 2 weeks in 40 patients with AMI within 12 hours after the onset of the symptom and who randomly received early treatment with either the ACE inhibitor imidapril or a placebo (20 patients in the imidapril group and 20 in the placebo group). The levels of serum ACE activity in the imidapril group decreased significantly (p < 0.01) 8 hours after the administration of imidapril, and the levels 24 hours after administration were significantly lower than those in the placebo group (3.6 +/- 0.6 IU/L vs 7.4 +/- 0.8 IU/L; p < 0.001). The plasma PAI activity increased gradually to peak levels 16 hours after the administration of imidapril and placebo. The levels in the placebo group decreased gradually but remained high during the study period. On the other hand, the levels of PAI activity in the imidapril group decreased rapidly and those 48 hours after administration were significantly lower than those in the placebo group (7.9 +/- 1.9 IU/ml vs 18.4 +/- 3.5 IU/ml; p < 0.01). The levels of left ventricular ejection fraction about 2 weeks after admission were significantly higher in the imidapril group than in the placebo group (65.9% +/- 2.5% vs 49.1% +/- 4.4%; p < 0.01). This study showed that imidapril, an ACE inhibitor, might be useful for the improvement of fibrinolytic function and left ventricular function in the acute phase of myocardial infarction. PMID- 9398111 TI - Role of nitric oxide in coronary vasomotion during handgrip exercise. AB - BACKGROUND: Endothelium-dependent modulation of coronary vasomotion during increased sympathetic tone remains unclear in normal and atherosclerotic human coronory arteries. METHODS AND RESULTS: We evaluated the role of endothelium derived nitric oxide in vasomotion during isometric exercise in normal subjects (n = 7) and in patients with coronary artery disease (CAD) (n = 10). Coronary blood flow and epicardial coronary artery diameter to the handgrip test were measured before and after intracoronary administration of 100 micromol/min of N(G)-monomethyl L-arginine (L-NMMA). Heart rate and aortic blood pressure increased during handgrip test. Handgrip test caused a significant dilation in the diameter of the epicardial coronary artery in normal subjects (9.9% +/- 3.9%, mean +/- SD) and in the diameter of smooth segments of patients with CAD (5% +/- 3.7%, p < 0.05 vs normal subjects). In contrast, the diameter of irregular segments in patients with CAD decreased during handgrip test (-9.8 +/- 3.9%). After L-NMMA, the epicardial coronary artery significantly increased during handgrip test compared with before L-NMMA in normal subjects. L-NMMA did not have any effect on handgrip test induced vasodilation in the smooth segments and vasoconstriction in the irregular segments in the patients with CAD. Handgrip test-induced increases in coronary blood flow did not change after L-NMMA in both groups. CONCLUSIONS: Nitric oxide does not play a major role in HNG-induced vasodilation in epicardial and microcirculatory vessels in normal human coronary circulation. Although the decreased release in nitric oxide may modulate the abnormal response of the epicardial coronary artery to handgrip test, this does not explain the paradoxic constrictive response from the depressed but still dilatory response in the patients with CAD. PMID- 9398112 TI - Hanshin-Awaji earthquake as a trigger for acute myocardial infarction. AB - On Jan. 17, 1995, the Hanshin-Awaji district was struck by the most destructive earthquake ever to occur in Japan. It is commonly believed that acute emotional stress such as that caused by an earthquake can trigger acute myocardial infarction (AMI). The objective of this study was to evaluate the effect of the quake stress on the onset of AMI in our district. The number of patients with AMI during the first 4 weeks after the quake increased by about 3.5-fold. The mean age of patients was 72.5 +/- 2.8 years, and the proportion of women (53%) was significantly greater than in the preceding years. The proportion of patients without prodromal angina pectoris was 53%. The mean post-traumatic stress disorder reaction index score (n = 14) was 40.1 +/- 4.1, which indicates a severe stress level. The mean score in the women (45.9 +/- 4.7; n = 7) was significantly higher than that in the men (34.3 +/- 6.4; n = 7). We concluded that after an earthquake, severe emotional stress can trigger AMI, more often than normal in women. PMID- 9398113 TI - Association of hemostatic factors with peripheral vascular disease. AB - Hemostatic risk factors have been well established in coronary artery disease but less well studied in peripheral vascular disease. The relationship of coagulation and fibrinolytic proteins to lower limb arterial occlusive disease and other vascular risk factors remains poorly defined. Fibrinogen, factor VII coagulant activity, von Willebrand factor (vWf) antigen, and plasminogen activator inhibitor-1 (PAI-1) activity were measured in 46 adult participants in the Arterial Disease Multiple Intervention Trial (ADMIT) and in 76 control subjects and related to ankle-brachial systolic pressure index (ABI), a measure of lower limb arterial stenosis. The primary inclusion criterion for the ADMIT study population was an average of two ABIs <0.85. Fibrinogen and PAI-1 in ADMIT subjects were significantly higher than in control subjects (331 +/- 52 mg/dl vs 273 +/- 46 mg/dl, p < 0.0001; 18.7 +/- 10 units/ml vs 13.5 +/- 8.9 units/ml, p < 0.04). There were significant correlations of fibrinogen with ABI, factor VII coagulant activity, and systolic and diastolic blood pressures; PAI-1 with body mass index and age; and factor VII coagulant activity with cholesterol levels. Logistic regression analysis, considering hemostatic variables and several known nonhemostatic risk factors of peripheral arterial disease, showed that fibrinogen and systolic blood pressure were independently associated with ABI status in this population. The results demonstrate a strong independent correlation between fibrinogen levels and the presence of lower limb arterial stenosis. PAI-1 levels were elevated in ADMIT participants, but multivariate analysis did not demonstrate an independent relationship between PAI-1 and ABI. PMID- 9398114 TI - Gravitational forces and bone metabolism. PMID- 9398115 TI - Association of gender and access to cadaveric renal transplantation. AB - Previous studies have revealed that females are less likely than males to receive a renal transplant, the most successful form of treatment of end-stage renal disease (ESRD). The purpose of this study was to determine whether the barrier is to inclusion on the transplant waiting list or to transplantation after being placed on the transplant waiting list. An existing data set was used that included data from the Michigan Kidney Registry, supplemented with data received from the Organ Procurement Agency of Michigan. White and black patients less than 65 years of age and starting ESRD treatment between January 1, 1984, and December 31, 1989, were included. Cox proportional hazards models were used to determine the effect of gender on (1) time to transplantation among all ESRD patients, (2) time from diagnosis of ESRD to inclusion on the transplant waiting list among all ESRD patients, and (3) time from inclusion on the waiting list to transplantation among those patients on the waiting list. Patients were censored at the time of living-related transplantation or death, and were monitored until December 31, 1989. In all, 5,026 incident ESRD patients were included in the study (44.3% female). Of these, 1,626 patients were included on the waiting list (40.1% female); 823 of these received a transplant (37.7% female). Adjusting for age, race, and diagnosis, females were 25% less likely to receive a cadaveric transplant than males (female to male relative rate ratio [RR], 0.75; P < 0.001). Females with ESRD aged 46 to 55 years and 56 to 65 years were 33% (RR, 0.67; P < 0.001) and 29% (RR, 0.71; P < 0.05) less likely to be included on the transplant waiting list, respectively, than their male counterparts. There was no difference in the rate of wait list inclusion among ESRD patients younger than 46 years. Females with ESRD who were included on the transplant waiting list were 26% (RR, 0.74; P < 0.001) less likely to receive a transplant than males on the waiting list. These results indicate that females are both less likely to be on the transplant waiting list (ages over 45 years) and, once on the list, less likely to receive a transplant (all ages) than males. Further study is necessary to determine the factors contributing to these important barriers to transplantation among females with ESRD. PMID- 9398116 TI - Factors determining the rate of referral, transplantation, and survival on dialysis in women with ESRD. AB - The determinants of referral for transplantation in women have not been well studied. Similarly, factors determining survival on dialysis and the rate of transplantation in women remain controversial. Women have been reported to have lower rates of transplantation than men, and black women have the lowest rates of all groups. We questioned whether black women were referred at lower rates than whites and if race and other socioeconomic factors predicted referral, rate of renal transplantation, and patient survival on dialysis. All women in Allegheny or Philadelphia counties in Pennsylvania initiating dialysis between January 1, 1990, and December 31, 1992, were eligible for this study. Information was requested by questionnaire from each dialysis unit in these areas. Of the 383 eligible patients, completed questionnaires were obtained for 276 (72%). Ninety three (54.7%) of the black patients and 57 (53.8%) of the white patients were referred for transplantation (P = 0.8). Declining the transplant option was the most common reason for nonreferral in both races. Patients with high school or greater education were approximately twice as likely to be referred than those with grade school educations (odds ratio [OR], 2.2; P = 0.04). Patients with coexisting illness were 67% (OR, 0.33; P = 0.004) less likely to be referred compared with patients with no other illnesses. Each additional year of age reduced the chances of being referred by 6% (OR, 0.94; P = 0.0001). Homemakers were 83% (P = 0.0008) and others 55% (P = 0.07) less likely to be referred compared with employed patients. Patients in Philadelphia County were 56% less likely to be referred compared with those in Allegheny County (P = 0.024). Race was not significantly associated with referral. Predictors of transplantation included age (RR, 0.96; confidence interval [CI], 0.93 to 0.99; P = 0.13), white race (RR, 2.2; CI, 1.3 to 4.0; P = 0.0053), presence of other illnesses (RR, 0.37; CI, 0.21 to 0.65; P = 0.0006), and employment status. White homemakers were 86% (RR, 0.14; CI, 0.03 to 0.6; P = 0.0082) less likely than those with other employment situations to receive a transplant. Factors predicting patient survival on dialysis included race, educational status, and presence of comorbid illnesses. White patients were approximately four times (RR, 3.7; CI, 1.7 to 8.1; P = 0.002) more likely to die than black patients. Patients with high school or greater education were 56% (RR, 0.44; CI, 0.2 to 0.92; P = 0.008) less likely to die than those with grade school education alone. Patients with at least one coexisting illness were approximately 1.7 times (RR, 1.68; CI, 1.1 to 2.4; P = 0.001) more likely to die than those without other illnesses. In summary, race was not a factor in referral for transplantation, but was predictive of transplantation and patient survival on dialysis. Socioeconomic factors such as educational status, age, and employment status were highly predictive of transplantation and long-term survival on hemodialysis. White homemakers unexpectedly received transplants less than any other group of dialysis patients. Further study is needed to determine why these potential transplant patients have declined or deferred transplantation. PMID- 9398117 TI - Minimizing racial disparity regarding receipt of a cadaver kidney transplant. AB - This report describes the impact of race on waiting list entry and receipt of a cadaver kidney transplant, after accounting for self-reported income, health and functional status, and patients' attitudes about dialysis and transplantation as treatment alternatives. Previous studies did not account for these race-related factors and therefore produced biased estimates of the impact of race on waiting list entry and receipt of a transplant. Data for this investigation came from a telephone survey of a national sample of 456 end-stage renal disease patients and from files maintained by the United Network for Organ Sharing and the Health Care Financing Administration. Proportional hazard models were estimated with these data. The results indicated that approximately 60% of the differences between black and white waiting list entry rates and 52% of the black-white differences in transplantation rates were due to race-related differences in socioeconomic status, health and functional status, severity of illness, biological factors, the existence of contraindications to transplantation, transplant center characteristics, and patients' attitudes about dialysis and transplantation. Potential ways to narrow racial differences further include better education about treatment alternatives for black patients, more vigorous efforts to obtain donor organs from minorities, continued research and thoughtful policy on the access-related impacts of United Network for Organ Sharing point system variances, and consolidation of some smaller waiting lists into larger regional lists. PMID- 9398118 TI - Renal transplantation in adults with autosomal recessive inheritance of hemolytic uremic syndrome. AB - When hemolytic uremic syndrome (HUS) is occasionally inherited in an autosomal recessive mode, this occurs mainly in infants and children. We describe four families in which two adult siblings were affected with HUS in each kindred. HUS first occurred between the ages of 19 to 36 years, and the intervals between the onset of HUS in each sibling pair ranged from 6 months to 6 years. None of the patients had a typical prodrome of bloody diarrhea, and one had a recurrence of HUS before transplantation. All eight patients developed renal failure requiring dialysis and transplantation, and seven patients received kidney transplants. Donor kidneys were from parents, siblings, and cadavers. The initial renal transplants were performed from 6 months to 6 years after the onset of the syndrome. HUS recurred in six of the seven patients 2 weeks to 6.5 years after transplantation regardless of the interval between the onset of HUS and transplantation, the origin of the allograft, or the use of cyclosporin A. The only marker for autosomal recessive HUS is the occurrence of the syndrome in a second sibling several months to many years after its occurrence in the proband. In patients with the autosomal recessive form of HUS, the risk for a recurrence in an allograft is high regardless of the source of the kidney. PMID- 9398119 TI - Cytomegalovirus and HLA-A, B, and DR locus interactions: impact on renal transplant graft survival. AB - Graft failure rates for renal transplantations performed between 1989 and 1994 and recorded in the US Renal Data System database were retrospectively evaluated for interactions between cytomegalovirus and HLA-A, B, and DR loci. Twelve significant interactions were observed. There were significantly greater risks of graft failure for the total effect of cytomegalovirus and donor or matched HLA DR9, recipient or matched HLA-B-51, and matched HLA-B13. We conclude that further study of renal transplants with these combinations of cytomegalovirus and HLA loci is needed to determine whether the observed interactions should be taken into consideration when matching donors with recipients. PMID- 9398120 TI - Intradialytic hypotension: is midodrine beneficial in symptomatic hemodialysis patients? AB - Symptomatic hypotension during hemodialysis is a disabling complication in end stage renal disease (ESRD) patients, especially in certain groups of patients who are at higher risk for this problem. Autonomic dysfunction is thought to play a significant role. We evaluated the efficacy of midodrine, an oral agent with selective alpha-adrenergic agonist activity used in the treatment of neurogenic orthostatic hypotension, on 10 hemodialysis patients with persistent intradialytic hypotension. The patients were given a dose of midodrine (mean dose, 5.5 mg; range, 5 to 10 mg) 30 minutes before each hemodialysis session. We compared blood pressure, pulse, body weight, and laboratory values for 10 consecutive dialysis sessions off and on midodrine therapy. There was a statistically significant improvement in lowest intradialytic systolic blood pressure (from 96.6 to 114.7 mm Hg; P < 0.001), lowest intradialytic diastolic blood pressure (from 53.2 to 59.0 mm Hg; P = 0.002), lowest intradialytic mean arterial pressure (from 67.7 to 77.6 mm Hg; P < 0.001), posthemodialysis systolic blood pressure (from 116.5 to 127.1 mm Hg; P < 0.001), posthemodialysis diastolic blood pressure (from 66.6 to 69.7 mm Hg; P = 0.040), and posthemodialysis mean arterial pressure (from 83.2 to 88.8 mm Hg; P = 0.001) after patients were placed on midodrine. There also was a small but statistically significant decrease in intradialytic pulse rate (from 86.3 to 81 beats/min; P = 0.021) and posthemodialysis pulse rate (from 87.4 to 81.7 beats/min; P = 0.024) after initiation of midodrine therapy. There was no significant difference in any of the prehemodialysis blood pressure measurements or pulse rate off or on midodrine therapy. The improvements in intradialytic and posthemodialysis blood pressure were associated with a uniform subjective improvement in symptoms associated with dialysis hypotension, such as cramps, fatigue, dizziness, and weakness. Other than scalp paresthesia in one patient, no adverse effects were noted. Our results suggest that the administration of a single dose of midodrine before hemodialysis is an effective therapy for intradialytic hypotension. A prospective trial with adequate patient numbers and long-term follow-up would be useful to evaluate this drug's efficacy and safety profile in patients with ESRD. PMID- 9398121 TI - Relationship between left ventricular hypertrophy, myocardial contractility, and load conditions in hemodialysis patients: an echocardiographic study. AB - Left ventricular hypertrophy (LVH) is common and is an independent cardiac risk factor in dialysis patients. The aim of this study was to assess hemodynamic determinants of LVH and, more particularly, the relationship between left ventricular mass, myocardial contractility, and load conditions. Eighty dialysis patients aged 51 +/- 15 years were prospectively studied by echocardiography. LVH was detected in 62 patients (78%). Left ventricular mass was significantly correlated to both end-diastolic volume (r = 0.54; P < 0.001) and end-systolic stress/end-systolic volume, an index of contractility (r = -0.66; P < 0.001), but not to systolic blood pressure or end-systolic stress, both indexes of afterload. Thus, in dialysis patients, the degree of LVH is significantly correlated with the severity of both left ventricular dilatation and contractile myocardial failure, but not with left ventricular afterload. PMID- 9398122 TI - Hemodialysis: an appropriate therapy in myeloma-induced renal failure. AB - To determine whether vigorous treatment with dialysis is of benefit to patients with myeloma-induced renal failure at presentation, we retrospectively reviewed outcomes in a group of patients diagnosed with multiple myeloma between January 1986 and September 1993. Increased age (P = 0.003), presence of renal impairment (P = 0.006), and failure to enter plateau phase (P < 0.001) were independently associated with shortened survival. However, there was no difference in outcome between patients with severe renal failure, those treated with dialysis, and those with milder renal impairment (median survival, 22 months in both groups), nor was reversibility of renal failure associated with any survival advantage. The lack of correlation between severity or reversibility of the renal failure and survival suggests that there may be characteristics of some patients or their underlying myeloma that are responsible both for renal impairment and for adverse prognosis. In this study, neither age, clinical stage, labeling index, nor response to treatment was able to account for the difference in outcome between patients with and without renal failure. The prolongation of life achieved in the dialysis patients such that their median survival was identical with that of the group with milder renal impairment was considered to represent a significant benefit to these patients and to justify the offer of dialysis to all patients requiring it. PMID- 9398123 TI - Patient's view of dialysis care: development of a taxonomy and rating of importance of different aspects of care. CHOICE study. Choices for Healthy Outcomes in Caring for ESRD. AB - Quality assessment efforts to enhance public accountability in dialysis care and to support provider efforts to improve care have lacked patient input. To develop brief patient evaluation or satisfaction surveys suitable for busy clinical settings, knowing patients' priorities can be helpful in deciding which aspects of care should be tracked. We conducted a study to identify salient attributes of dialysis care and to rank the importance of these attributes from the perspective of dialysis patients. We analyzed the content of patient focus group transcripts to characterize dialysis care from the patients' perspective. We then surveyed 86 patients to determine how patients would rank the importance of each aspect to quality of dialysis care. The 18 broad aspects of care identified in the focus group included: (1) care provided by nephrologists, (2) care provided by other physicians (nonnephrologists), (3) care provided by dialysis center nurses, (4) care provided by social workers and psychologists, (5) care provided by dieticians, (6) clergy, (7) care provided by technicians and physician assistants/nurse practitioners, (8) care provided by dialysis center staff in general, (9) supplies, (10) treatment choice and effectiveness, (11) patient education and training, (12) self-care, (13) dialysis machines, (14) unit environment and policies, (15) cost containment, (16) billing, (17) cost of care, and (18) health outcomes. Items ranked in the top 10 by both groups of patients included issues related to nephrologists, other doctors, nurses, and patient education and training. Compared with hemodialysis patients, peritoneal dialysis patients gave higher ratings to hospital doctors' and nurses' attention to cleanliness when working with access sites, how correct the nephrologist's instructions to patients are, whether emergency room doctors check with nephrologists, the amount of information patients get about their diet, and how well nurses answer patients' questions. Patients value certain aspects of dialysis care highly, and these aspects differed in some respects for the relatively small number of hemodialysis and peritoneal dialysis patients studied. Construction of brief questionnaires for quality assessment and assurance requires thoughtful consideration of what questions to include. Knowing patients' priorities regarding the most important aspects of care that have high potential for dissatisfaction may be helpful to continuous quality improvement of end-stage renal disease care. PMID- 9398124 TI - Supraclavicular approach to the subclavian/innominate vein for large-bore central venous catheters. AB - Infraclavicular and internal jugular catheterization are commonly used techniques for hemodialysis access, but may at times be impeded in patients whose anatomy makes cannulation difficult. In an effort to enlarge the spectrum of alternative access sites, we evaluated the supraclavicular approach for large-bore catheters. During an 18-month period we prospectively collected data on success rate and major and minor complications of the supraclavicular access for conventional dialysis catheters as well as Dacron-cuffed tunneled devices in 175 adult patients admitted for various extracorporeal therapies and bone marrow transplantation. Two hundred eight large-bore catheters (99 conventional dialysis catheters, 63 semirigid tunneled Dacron-cuffed catheters, and 46 Hickman catheters) were successfully placed in 164 patients (success rate, 93.8%), 58 (33.1%) of whom had been previously catheterized. Complications included pneumothorax (one patient), arterial puncture (seven patients), and puncture of the thoracic duct (two patients) without sequelae. Postinsertional chest radiographs demonstrated impressive coaxial lie of most catheters. Catheter malpositions occurred only sporadically (1%). Difficulty of introducing the catheter via a placed sheath was rarely observed. There was no clinically significant evidence of catheter-induced venous thrombosis or stenosis. We conclude that the supraclavicular route is an easy and safe first approach for large-bore catheters, as well as a useful alternative to traditional puncture sites for precatheterized and anatomically problematic patients. PMID- 9398125 TI - Evidence that serum phosphate is independently associated with serum PTH in patients with chronic renal failure. AB - There has been controversy regarding the initial pathogenic events involved with the hyperparathyroidism of chronic renal failure (CRF). Low serum levels of 1,25 dihydroxyvitamin D in uremic patients are postulated by some as having a role in permitting higher parathyroid hormone (PTH) secretion. However, recent animal and in vitro studies strongly suggest that phosphate has a direct effect on parathyroid cells to enhance PTH secretion. To evaluate the relationships among serum phosphate, calcium, PTH, and 1,25-dihydroxyvitamin D in uremic humans, we performed a cross-sectional analysis of 84 patients with varying levels of CRF. Using stepwise regression analysis after adjusting for multiple comparisons, we found that serum phosphate correlated directly with serum PTH (r = 0.62, P < 0.01) in patients with mild to moderate CRF (creatinine < or = 3.0 mg/dL), independent of serum calcium and 1,25-dihydroxyvitamin D levels. In patients with more severe renal failure (creatinine > 3.0 mg/dL), only the serum calcium correlated with serum PTH (r = -0.47, P < 0.01). While serum 1 ,25 dihydroxyvitamin D showed no correlations with PTH, phosphate, or calcium at any stage of renal failure, the mean 1,25-dihydroxyvitamin D level in patients with mild CRF was lower than that in age-matched controls (24 +/- 3 pg/mL v 37 +/- 2 pg/mL; P < 0.01), suggesting that low 1,25-dihydroxyvitamin D was permissive for enhanced PTH secretion. These data demonstrate an independent association of serum phosphate with PTH in patients with CRF and suggest that phosphate may directly enhance PTH secretion in this setting. This study supports recent animal studies showing a direct parathyroid cell effect of phosphate on PTH secretion. PMID- 9398126 TI - Evaluation of RBC ferritin and reticulocyte measurements in monitoring response to intravenous iron therapy. AB - Intravenous (IV) iron therapy can reduce erythropoietin (EPO) requirements in dialysis patients. Monitoring this response accurately is difficult. Estimation of red blood cell ferritin (RBCFer) and reticulocyte indices may give additional valuable information about iron availability to the erythroblasts (erythron). We evaluated the use of RBCFer, mean hemoglobin content of reticulocytes (CHr), and mean hemoglobin concentration of reticulocytes (CHCMr) in a prospective, nonblinded study of 22 hemodialysis patients (16 men and six women with a mean age of 62 years [range, 24 to 80 years]). All patients had an initial serum ferritin of < or = 60 microg/L. Patients with features known to produce EPO resistance and underlying bleeding/hematologic disorders were excluded. Patients were established on subcutaneous EPO and given IV iron therapy. The mean hemoglobin level remained constant throughout the study (P = 0.087). Serum ferritin and RBCFer increased significantly (P < 0.001 and 0.015, respectively) while a reduction in transferrin saturation became significant at the end of the study (P = 0.0047). A sharp increase in reticulocytes occurred in the first 14 days after commencement of IV iron, and there was an initial decrease in the percentage of hypochromic RBCs. An early decline in RBCFer was apparent. CHr increased with IV iron, indicative of increased iron supply to the developing erythron. Measurement of RBCFer and CHr provide evidence of increased iron supply for erythropoiesis during IV iron therapy. These measures help identify patients with functional iron deficiency and allow more accurate monitoring of response to IV iron therapy. PMID- 9398127 TI - Oncotic pressure and edema formation in hypoalbuminemic HIV-infected patients with proteinuria. AB - Human immunodeficiency virus nephropathy (HIVN) continues to challenge nephrologic consultative services at major urban institutions. Although noted in the literature, the decreased incidence of peripheral edema in HIVN has been unexplained to date. In HIV patients, total proteins frequently are found to be elevated due to an elevated globulin fraction. The impact that plasma proteins, specifically globulins, have on the total oncotic pressure has not been reported in HIVN, but may play a role in the paucity of edema noted in this proteinuric population. To evaluate the contributions of serum globulin to the total oncotic pressure and the presence or absence of edema in HIVN, we randomly selected 27 patients with proteinuria greater than 2.5 g/24 hr and serum albumin less than 3.1 g/dL from patients presenting to the nephrology outpatient clinic at the University of Miami/Jackson Memorial Hospital. Seventeen of the patients (63%) had a known diagnosis of HIV infection (group 1). These patients were subdivided into two subgroups: those presenting with clinically evident edema on physical examination (n = 7 [41%]; group 1A) and those who had an absence of edema (n = 10 [59%]; group 1B). Conversely, group 2 comprised 10 patients without known HIV infection, of whom six (60%) had edema (group 2A) and four (40%) did not (group 2B). Blood pressures were noted, and mean arterial pressure was calculated using standard formulas. Serum albumin, serum total proteins, and urine total proteins were measured using standard laboratory methods. Oncotic pressures for albumin (alpha), globulin (beta), and total protein (c) were calculated using the following formula: COPpl = alpha(2.8c + 0.18c2 + 0.012c3) + beta(0.9c + 0.12c2 + 0.004c3). We used Student's t-test to analyze the data. There is no significant difference between the albumin concentrations of HIV patients without edema (group 1B) and non-HIV patients with edema (group 2A), with mean concentrations of 2.3 +/- 0.1 g/dL versus 2.3 +/- 0.15 g/dL, respectively (P = NS). Group 1B, however, has a total oncotic pressure of 17.1 +/- 1.5 mm Hg, whereas both groups with edema (groups 1A and 2A) have statistically significant lower total oncotic pressures (12.1 +/- 2.3 mm Hg and 12.9 +/- 1.1 mm Hg, respectively; P < 0.05). The globulin oncotic pressures may account for some of the differences in total oncotic pressures, being significantly higher for those patients without edema in group 1B compared with group 2A (7.1 +/- 0.9 mm Hg v 3.9 +/- 0.4 mm Hg, respectively; P < 0.05). In patients with HIV, however, the presence or absence of edema is mandated by albumin concentration because both groups have similar globulin concentrations (group 1A 3.1 +/- 0.1 g/dL v group 1B 3.8 +/- 0.3 g/dL; P = NS). Mean arterial pressure does not play a role in edema formation in this study because the HIV patients without edema had the higher blood pressures (group 1B 97.8 +/- 4.7 mm Hg v group 2A 84.7 +/- 5.5 mm Hg; P < 0.05). We conclude that globulins play an important role in maintaining oncotic pressure in low albumin states. HIVN patients with increased serum immune globulin may benefit from higher globulin oncotic pressure, delaying the onset of clinical edema in the setting of proteinuria. PMID- 9398128 TI - Factors contributing to the degree of polyuria in a patient with poorly controlled diabetes mellitus. AB - Polyuria due to a glucose-induced osmotic diuresis is common in patients with hyperglycemia. This diuresis usually abates when the plasma glucose level approaches its renal threshold; the usual time course is less than 8 hours after commencing therapy. A 69-year-old man with non-insulin-dependent diabetes mellitus maintained hyperglycemia (540 mg/dL) and polyuria (4.7 L/24 hr) for 40 hours. Because there was no external supply of glucose, a balance study was conducted between the third and 40th hour after commencing treatment. In this interval, the overall concentration of glucose in the urine was less than 100 mmol/L and the urine osmolality was 378 mOsm/kg H2O. To evaluate the expected composition of the urine during a glucose-induced osmotic diuresis, urine was analyzed in normal rats infused with glucose plus urea and in untreated BB diabetic rats (plasma glucose and urea similar to that in our patient) as well as in 29 patients with hyperglycemia and polyuria. Glucose accounted for 60% of the urinary osmoles in rats and humans. Two subgroups of patients had a much lower urine glucose: one had an impaired concentrating ability (n = 6) and the other had an increased rate of renal glucose reabsorption (n = 5). In conclusion, in polyuria caused by hyperglycemia, the urine glucose should be 300 to 400 mmol/L with normal renal function. In the case we report, both the concentration of glucose and its excretion rate were much lower than expected with steady-state hyperglycemia (540 mg/dL) due to the high rate of excretion of NaCl, a concentrating defect, and excessive renal reabsorption of glucose. PMID- 9398129 TI - Secondary collapsing glomerulopathy associated with Loa loa filariasis. AB - This is the first case of nonprimary collapsing focal segmental glomerulosclerosis (FSGS) associated with Loa loa filariasis. Loa loa micofilariae were detected on a blood smear after a patient presented with nephrotic syndrome (NS), microhematuria, and renal failure. The renal biopsy showed a collapsing glomerulopathy variant of FSGS. Microfilariae also were identified in renal microvasculature, including the afferent arterioles and the glomerular and peritubular capillaries. PMID- 9398131 TI - Xanthogranulomatous pyelonephritis in a renal allograft recipient. AB - Xanthogranulomatous pyelonephritis rarely occurs in renal allografts. This is the fifth reported case. Diagnosis was made by renal biopsy, which is usually performed to evaluate an elevated serum creatinine. Associated patient symptomology is nonspecific, and graft imaging with ultrasonography and computed tomography was not helpful as it would be with native kidney xanthogranulomatous pyelonephritis. Successful treatment with antibiotics may depend on the serum creatinine at presentation. Prognosis, therefore, is guarded, with a common outcome of irreversible renal dysfunction. PMID- 9398130 TI - Leptospirosis: an ignored cause of acute renal failure in Taiwan. AB - Leptospirosis, caused by a spirochete, is the most common zoonosis in domestic or wild animals. Animals excrete infected urine in soil or water and may cause human infections through abrased wound, mucosa, conjunctiva, or by swallowing contaminated water. Clinical presentations of leptospirosis are mostly subclinical. Five to ten percent of leptospirosis are fatal, causing fever, hemorrhage, jaundice, and acute renal failure (Weil's syndrome). Leptospirosis has been ignored as a cause of acute renal failure in Taiwan. We report two patients with leptospirosis who presented with high fever, abdominal pain, jaundice, and acute renal failure. Patient 1 died on day 12 of admission of multiple organ failure associated with pancytopenia, hypogammaglobulinemia, and reactive hemophagocytosis. Leptospirosis was recognized after death. Patient 2 was admitted with similar presentations 2 weeks later. Penicillin and doxycycline were given early in the course, and azotemia, jaundice, respiratory failure, and aseptic meningitis gradually improved. Renal biopsy showed interstitial nephritis. Several tubular clearance tests showed proximal tubular defect with severe bicarbonate wasting (FeHCO3- 20.9%) and incomplete type II renal tubular acidosis without affecting the distal nephron. After 80 days of treatment, this patient was discharged with recovery of conscious level and renal function. This is the first leptospirosis patient with detailed tubular functional and morphological studies of the kidney. Diagnosis of leptospirosis was made by microscopic agglutination test (MAT) for antibody to leptospira and by polymerase chain reaction (PCR) for leptospira DNA in blood and urine (interrogans serogroup australis in case 1 and Leptospira borgpetersenii serogroup ballum in case 2). Because active surveillance has resulted in 13 cases diagnosed as leptospirosis islandwide thereafter, underestimation and ignorance of leptospirosis as a cause of acute renal failure may occur in Taiwan. Therefore, an area with a low leptospirosis incidence may actually have a very high incidence. Leptospirosis should be suspected in febrile patients with jaundice and renal failure when pathogens cannot be identified by traditional culture for microorganisms. PMID- 9398132 TI - Systemic thrombolysis for acute myocardial infarction in a renal transplant recipient. AB - This study describes the successful management of an acute myocardial infarction occurring in a renal transplant recipient with thrombolytic therapy. Although primary coronary angioplasty has been addressed as an alternative therapeutic approach, this approach raises concern for angiography-related contrast media renal toxicity. However, pharmacological therapy with thrombolytics is effective and relatively safe and should be considered as the first-choice treatment in today's clinical setting. PMID- 9398133 TI - Renal artery dissection causing renal infarction in otherwise healthy men. AB - Arterial dissection is usually associated with pathological states such as malignant hypertension, severe atherosclerosis, severe trauma, Marfan syndrome, or Ehlers-Danlos syndrome. However, we report three cases in which renal artery dissection occurred in otherwise healthy, normotensive men. In two cases, the onset of symptoms of renal artery dissection was coincident with an unusual degree of physical activity. In the third case, the symptoms occurred while the patient was sitting but during a stressful business meeting. In each case, the patient experienced severe unilateral flank pain. Urolithiasis was suspected, but intravenous pyelography showed only ipsilateral impaired renal cortical perfusion, and the urinalyses showed no hematuria. The diagnosis of renal artery dissection was established by arteriography in two cases and by nephrectomy in one case. The latter case showed fibromuscular dysplasia by arteriography performed after the nephrectomy. The other two cases showed no evidence of fibromuscular dysplasia. We conclude that spontaneous renal artery dissection can occur in otherwise healthy individuals. Our experience and the reports of others indicate that this condition occurs mainly in men, conservative (nonsurgical) management is generally indicated, and the long-term prognosis is generally excellent. In some patients, an unusual degree of physical exertion might be the cause of renal artery dissection. PMID- 9398134 TI - Influence of gender and race on therapeutic options for ESRD patients. PMID- 9398135 TI - National Kidney Foundation report on dialyzer reuse. Task Force on Reuse of Dialyzers, Council on Dialysis, National Kidney Foundation. AB - The Council on Dialysis of the National Kidney Foundation convened an expert panel to evaluate the current practice and literature related to the reuse of hemodialyzers. The panel reviewed and evaluated literature related to reuse since the last report of the National Kidney Foundation recommendations on reuse was published in 1988. The group sought to develop a consensus concerning the effect of reuse of hemodialyzers on mortality; the efficiency of delivered hemodialysis when reused hemodialyzers are used in the clinical setting; the clinical effects of reused dialyzers as compared with dialyzers not reused on intradialytic symptoms; infections in patients using reused dialyzers; and the effect of reused dialyzers on complement activation, cytokine production, and beta2-microglobulin metabolism and clearance. In addition, the panel reviewed the literature on the potential toxicity of germicides used in the processing of dialyzers for reuse as well as recent changes in federally mandated regulations concerning labeling of dialyzers for reuse, the monitoring of the reuse process, and the effectiveness of reused dialyzers to achieve a prescribed delivered clearance as estimated by urea kinetic modeling or by percent urea reduction. The National Kidney Foundation takes no position for or against dialyzer reuse. The principal reason for the practice of reuse is economical. In view of the uncertainties related to the safety and biological impact of reuse procedures, the task force recommends that a full discussion of the issue of reuse and its potential beneficial and detrimental effects be undertaken with each patient. There is no conclusive evidence to substantiate the notion that either morbidity or mortality associated with single use or reuse is different. Microbial contamination of the water used for dialyzer reprocessing increases patient morbidity. The chemical quality of water used for dialyzer reprocessing should, at least, fall within the same standards as those recommended for product water intended for hemodialysis. Dialyzers should not be reprocessed from patients who have tested positive for hepatitis B surface antigen. The effects of reprocessing high-flux dialyzers on beta2-microglobulin clearance are dependent on the reprocessing technique, the number of reuses, and the nature of the dialyzer membrane used. There are insufficient data on the effects of reuse on beta2-microglobulin behavior to make uniform recommendations. Untoward effects of reused dialyzers may still occur in spite of rigorous adherence to the AAMI guidelines. For example, use of the total cell volume method for assessing changes in small molecule clearances will not show the loss of performance attributable to dialysate shunting. For this reason, the measurement of Kt/V for urea as recommended by the AAMI or the determination of the urea reduction ratio (URR) is strongly recommended at least monthly to gauge the adequacy of the dialysis procedure. Given the significant fall in dialyzer efficiency for urea removal that can occur after repeated uses of a dialyzer, dialysis prescriptions in units practicing reuse should be designed to deliver a Kt/V or URR value that exceeds the dose used for patients treated with single-use dialyzers to make allowance for any possible reuse-induced reduction in dialyzer efficiency. Technicians and other personnel responsible for the reprocessing of dialyzers should receive proper training. These health care providers should be certified in reprocessing by an examining body so that professional competency can be assured. PMID- 9398136 TI - Cyclophosphamide therapy of severe lupus nephritis. PMID- 9398137 TI - Sickle cell nephropathy during the postpartum period in a patient with SLE. PMID- 9398138 TI - Expression and function of calcineurin in the mammalian nephron: physiological roles, receptor signaling, and ion transport. AB - Protein phosphorylation is central to the regulation of sodium transport and other cellular processes in the nephron. Complex interactions between protein kinases and phosphatases catalyze the reversible phosphorylation of ion transporting proteins on the apical and basolateral surfaces of renal epithelia. Although the role of protein kinases in regulating sodium transport has been extensively studied, the function of phosphatases in the nephron is less well understood. Calcineurin is a serine-threonine phosphatase that was shown to be the target of cyclosporin A (CsA) and FK-506 in lymphocytes. Calcineurin exists in the cytosol as a heterotrimeric protein composed of an alpha-catalytic subunit, beta-regulatory subunit, and calmodulin; its activity depends on calcium and calmodulin. Three isoforms of the alpha-subunit (alpha-1, alpha-2, alpha-3) and two isoforms of the beta-subunit (beta-1 and beta-2) of calcineurin have been identified. In proximal tubules, alpha-1 isoforms are predominant and exceed alpha-2 expression by fourfold. In the CCD, alpha-1 and alpha-2 expression are approximately equal, whereas alpha-2 subunit expression is greatest in medullary thick ascending limbs (mTAL). Alpha-3 was not detected in any nephron segment. Calcineurin phosphatase activity in the proximal tubule is approximately 10-fold higher than in the connecting tubules (CNT), cortical collecting ducts (CCD), or the mTAL. Protein phosphatases 1 and 2a are also expressed in CCD, and only protein phosphatase 1 can be detected in the proximal tubule. Calcineurin influences basal and stimulated Na/ K-ATPase activity in the proximal and distal nephron. In the CCD, CsA or FK-506 decrease Na/K-ATPase activity by 35% and 85%, respectively; Na/K-ATPase activity in mTAL is decreased by 53% and 56%. Activation of membrane receptors, including adrenergic, dopamanergic, and angiotensin I receptors, also regulates Na/K-ATPAse activity through processes that involve calcineurin. Lastly, steroid hormones including glucocorticoids and mineralocorticoids appear to activate calcineurin phosphatase activity. The mechanism is independent of transcription and appears to involve mechanisms involving heat shock proteins associated with the steroid receptor complex. PMID- 9398139 TI - An evidence-based approach to earlier initiation of dialysis. AB - The objective was to review evidence addressing the optimal time to initiate dialysis treatment. The database was derived from an evidence-based review of the medical literature and from the Canada-United States peritoneal dialysis study. The publications were divided into (1) those addressing the clinical impact of early versus late referral to a dialysis program; (2) those evaluating the association between residual renal function at initiation of dialysis and the concurrent nutritional status; (3) those evaluating the association between residual renal function at initiation of dialysis and subsequent clinical outcomes, including patient survival. There were five studies evaluating early versus late referral, three cohort design and two case-control design. Late referrals had worse outcomes than early referrals. The former had more serious comorbidity and many had been noncompliant with follow-up. The latter were more likely to have hereditary renal disease. Renal function was slightly worse at initiation among those referred late. Three studies addressed the association between renal function at initiation of dialysis and concurrent nutritional status. Two showed decreased protein intake with diminished glomerular filtration rate (GFR). Poor nutritional status is associated with decreased patient survival among both incident and prevalent dialysis patients. The third study reported excellent patient survival among patients with late initiation of dialysis. These patients had received a supplemented low-protein diet and were not malnourished at initiation of dialysis. Three groups have studied the association between GFR at initiation of dialysis and clinical outcomes. Decreased GFR at initiation of dialysis is associated with a increased probability of hospitalization and death. None of these studies has used the rigorous randomized clinical trial design, and they are therefore subject to bias. Referral time bias, comorbidity, patient compliance, and starting time bias are potential confounders. A randomized clinical trial is required to resolve this important issue. However, there is sufficient evidence to justify initiation of dialysis at a Ccr of 9 to 14 mL/min if there is any clinical or laboratory evidence of malnutrition. PMID- 9398140 TI - Achieving target hematocrit in dialysis patients: new concepts in iron management. AB - The management of anemia in dialysis patients involves a comprehensive understanding of the role of erythropoietin deficiency and of the importance of adequate available iron. It is clear that iron and recombinant human erythropoietin (rHuEPO) in concert allow the clinician to achieve a given target hematocrit in dialysis patients. By first repleting and then maintaining iron stores, and with an appreciation of the concept of functional iron deficiency, the nephrologist can achieve target hematocrits with the lowest necessary dose of rHuEPO. Iron repletion and maintenance is difficult to achieve with oral iron, and parenteral iron is needed in most cases. New protocols for ongoing parenteral maintenance therapy with iron dextran or iron gluconate, a form of iron likely to be available soon in the United States, should lead to achievement of target hematocrits in a greater number of patients and be cost-effective in improving patient outcomes. PMID- 9398141 TI - Reticulocyte hemoglobin content predicts functional iron deficiency in hemodialysis patients receiving rHuEPO. AB - Early detection of iron sufficiency at the level of the erythropoietic cell is necessary to optimize management of uremic anemia with recombinant human erythropoietin (rHuEPO). "Absolute" and "functional" iron deficiency are the most important factors causing resistance to administered rHuEPO. Transferrin saturation and serum ferritin measurements have been noted to be insensitive and inaccurate measures to detect functional iron deficiency. Recently, the reticulocyte hemoglobin content (CHr) has been shown to be a sensitive and specific indicator of functional iron deficiency in nondialysis patients treated with rHuEPO. The purpose of this study is to compare CHr with currently used indices of iron sufficiency in rHuEPO-treated hemodialysis (HD) patients. In study 1, 364 stable HD patients were studied at two outpatient dialysis centers. CHr was normally distributed, with a mean value of 28.3 pg, and was consistent over two consecutive monthly samples in each center. CHr was weakly but consistently correlated with transferrin saturation and serum ferritin. CHr and reticulocyte number were inversely correlated with red blood cell (RBC) number, suggesting that the erythropoietic stimulus of routinely administered rHuEPO may have resulted in functional iron deficiency. Month-to-month changes in CHr correlated weakly with changes in serum iron and percent transferrin saturation, but not at all with changes in serum ferritin. When we analyzed those patients with baseline CHr less than 26 pg, a level strongly suggestive of functional iron deficiency, these correlations strengthened, and in addition, month-to-month changes in CHr correlated strongly and directly with concomitant changes in RBC count, hemoglobin, and hematocrit, suggesting that rising CHr was indicative of an erythropoietic response. In study 2, 79 patients received a single-dose infusion of 500 mg iron dextran. After intravenous iron, CHr rose within 48 hours, peaked at 96 hours, and then fell toward baseline. Patients who were iron deficient by standard measures (serum ferritin < 100 ng/mL or transferrin saturation less than 20%) had a greater and a sustained CHr response to intravenous iron dextran. A CHr less than 28 pg at baseline predicted functional iron deficiency, defined as a corrected reticulocyte increase of greater than 1% to iron dextran, more accurately than transferrin saturation, ferritin, or their combination. Eighty-two percent of individuals who were iron deficient at baseline responded to intravenous iron with an increase in CHr of greater than 2 pg. Sixty percent of patients who were iron sufficient by usual iron indices also responded to intravenous iron with a CHr rise of greater than 2 pg, suggesting that they were, in fact, functionally iron deficient despite "normal" conventional iron parameters. We conclude that CHr may be a more sensitive marker of functional iron deficiency in rHuEPO-treated hemodialysis patients than percent transferrin saturation and ferritin, particularly in those with "normal" conventional iron parameters. PMID- 9398142 TI - Acute phase proteins and peritoneal dialysate albumin loss are the main determinants of serum albumin in peritoneal dialysis patients. AB - Hypoalbuminemia predicts mortality in dialysis patients. It has been postulated that hypoalbuminemia in the dialysis population is a consequence of poor protein intake resulting from inadequate dialysis. To establish the cause of hypoalbuminemia in a group of 27 patients on peritoneal dialysis (PD), we determined the relationship between serum albumin concentration and a group of parameters including dialysis dose delivered (Kt/V), normalized protein catabolic rate (PCRn), transperitoneal and urinary albumin losses, and the serum concentration of two acute-phase proteins, C-reactive protein (CRP), and serum amyloid A (SAA). Serum albumin concentration could be predicted by a combination of transperitoneal albumin loss and either the serum concentration of CRP or of SAA. There was no relationship between weekly Kt/V or PCRn and serum albumin concentration. CRP and SAA significantly correlated with one another, but neither correlated with transperitoneal albumin losses. Hypoalbuminemia in PD patients is a consequence of transperitoneal albumin losses and of the acute phase response. PMID- 9398143 TI - The kidney: an unwilling accomplice in syndrome X. AB - The ability of insulin to stimulate glucose disposal by muscle varies widely within the population at large. Individuals with muscle insulin resistance develop type 2 diabetes if they cannot compensate for this defect by secreting large amounts of insulin. Although this philanthropic effort on the part of the pancreatic B-cell may prevent gross decompensation of glucose homeostasis, it renders such individuals at increased risk to develop a cluster of abnormalities (syndrome X) associated with coronary heart disease. Although the kidney is not considered to be an insulin sensitive tissue, two features of syndrome X, hyperuricemia and hypertension, are likely to be dependent on the retention of normal insulin action on the kidney. More specifically, there is evidence to support the hypothesis that elevated plasma insulin concentrations may enhance renal sodium retention and decrease urinary uric acid clearance. As such, it is possible that a normal kidney response to the compensatory hyperinsulinemia associated with insulin resistance in nondiabetic subjects contributes to the development of hyperuricemia and hypertension in such individuals. PMID- 9398144 TI - Ambulatory blood pressure monitoring in dialysis patients. PMID- 9398145 TI - Determinants of lipid profile in renal transplant recipients. PMID- 9398146 TI - Androgens, erythropoietin, iron stores, and lipoprotein (a) in hemodialysis patients. PMID- 9398147 TI - Modulation of tryptophan environment in membrane-bound melittin by negatively charged phospholipids: implications in membrane organization and function. AB - Melittin is a cationic hemolytic peptide isolated from the European honey bee, Apis mellifera. Since the association of the peptide in the membrane is linked with its physiological effects, a detailed understanding of the interaction of melittin with membranes is crucial. We have investigated the interaction of melittin with membranes of varying surface charge in the context of recent studies which show that the presence of negatively charged lipids in the membrane inhibits membrane lysis by melittin. The sole tryptophan residue in melittin has previously been shown to be critical for its hemolytic activity. The organization and dynamics of the tryptophan residue thus become important to understand the peptide activity in membranes of different charge types. Wavelength-selective fluorescence was utilized to monitor the tryptophan environment of membrane-bound melittin. Melittin exhibits a red edge excitation shift (REES) of 5 nm when bound to zwitterionic membranes while in negatively charged membranes, the magnitude of REES is reduced to 2-3 nm. Further, wavelength dependence of fluorescence polarization and near-UV circular dichroism spectra reveal characteristic differences in the tryptophan environment for melittin bound to zwitterionic and anionic membranes. These studies are supported by time-resolved fluorescence measurements of membrane-bound melittin. Tryptophan penetration depths for melittin bound to zwitterionic and anionic membranes were analyzed by the parallax method [Chattopadhyay, A., and London, E. (1987) Biochemistry 26, 39-45] utilizing differential fluorescence quenching obtained with phospholipids spin labeled at two different depths. Our results provide further insight into molecular details of membrane lysis by melittin and the modulation of lytic activity by negatively charged lipids. PMID- 9398148 TI - Chemical shift mapping of the RNA-binding interface of the multiple-RBD protein sex-lethal. AB - The Drosophila protein Sex-lethal (Sxl) contains two RNP consensus-type RNA binding domains (RBDs) separated by a short linker sequence. Both domains are essential for high-affinity binding to the single-stranded polypyrimidine tract (PPT) within the regulated 3' splice site of the transformer (tra) pre-mRNA. In this paper, the effect of RNA binding to a protein fragment containing both RBDs from Sxl (Sxl-RBD1 + 2) has been characterized by heteronuclear NMR. Nearly complete (85-90%) backbone resonance assignments have been obtained for unbound and RNA-bound states of Sxl-RBD1 + 2. A comparison of amide 1H and 15N chemical shifts between free and bound states has highlighted residues which respond to RNA binding. The beta-sheets in both RBDs (RBD1 and RBD2) form an RNA interaction surface, as has been observed in other RBDs. A significant number of residues display different behavior when comparing RBD1 and RBD2. This argues for a model in which RBD1 and RBD2 of Sxl have different or nonanalogous points of interaction with the tra PPT. R142 (in RBD2) exhibits the largest chemical shift change upon RNA binding. The role of R142 in RNA binding was tested by measuring the Kd of a mutant of Sxl-RBD1 + 2 in which R142 was replaced by alanine. This mutant lost the ability to bind RNA, showing a correlation with the chemical shift difference data. The RNA-binding affinities of two other mutants, F146A and T138I, were also shown to correlate with the NMR observations. PMID- 9398150 TI - Mapping the interfacial binding surface of human secretory group IIa phospholipase A2. AB - Human secretory group IIa phospholipase A2 (hIIa-PLA2) contains a large number of prominent cationic patches on its molecular surface and has exceptionally high affinity for anionic surfaces, including anionic membranes. To identify the cationic amino acid residues that support binding of hIIa-PLA2 to anionic membranes, we have performed extensive site-directed mutagenesis of this protein and measured vesicle binding and interfacial kinetic properties of the mutants using polymerized liposomes and nonpolymerized anionic vesicles. Unlike other secretory PLA2s, which have a few cationic residues that support binding of enzyme to anionic membranes, interfacial binding of hIIa-PLA2 is driven in part by electrostatic interactions involving a number of cationic residues forming patches on the putative interfacial binding surface. Among these residues, the amino-terminal patch composed of Arg-7, Lys-10, and Lys-16 makes the most significant contribution to interfacial adsorption, and this is supplemented by contributions from other patches, most notably Lys-74/Lys-87/Arg-92 and Lys 124/Arg-127. For these mutants, complete vesicle binding occurs in the presence of high vesicle concentrations, and under these conditions the mutants display specific activities comparable to that of wild-type enzyme. These studies indicate that electrostatic interactions between surface lysine and arginine residues and the interface contribute to interfacial binding of hIIa-PLA2 to anionic vesicles and that cationic residues closest to the opening of the active site slot make the most important interactions with the membrane. However, because the wild type binds extremely tightly to anionic vesicles, it was not possible to exactly determine what fraction of the total interfacial binding energy is due to electrostatics. PMID- 9398151 TI - Differential stabilization of the three FMN redox forms by tyrosine 94 and tryptophan 57 in flavodoxin from Anabaena and its influence on the redox potentials. AB - Flavodoxins are electron transfer proteins that carry a noncovalently bound flavin mononucleotide molecule as the redox-active center. The redox potentials of the flavin nucleotide are profoundly altered upon interaction with the protein. In Anabaena flavodoxin, as in many flavodoxins, the flavin is sandwiched between two aromatic residues (Trp57 and Tyr94) thought to be implicated in the alteration of the redox potentials. We have individually replaced these two residues by each of the other aromatic residues, by alanine and by leucine. For each mutant, we have determined the redox potentials and the binding energies of the oxidized FMN--apoflavodoxin complexes. From these data, the binding energies of the semireduced and reduced complexes have been calculated. Comparison of the binding energies of wild-type and mutant flavodoxins at the three redox states suggests that the interaction between Tyr94 and FMN stabilizes the apoflavodoxin- FMN complex in all redox states. The oxidized and semireduced complexes are, however, more strongly stabilized than the reduced complex, making the semiquinone/hydroquinone midpoint potential more negative in flavodoxin than in unbound FMN. Trp57 also stabilizes all redox forms of FMN, thus cooperating with Tyr94 in strong FMN binding. On the other hand, Trp57 seems to slightly destabilize the semireduced complex relative to the oxidized one. Finally, we have observed that reduction of mutants lacking Trp57 is slow relative to that of wild-type or mutants lacking Tyr94, which suggests that Trp57 could play a role in the kinetics of flavodoxin redox reactions. PMID- 9398152 TI - pH and temperature-induced molten globule-like denatured states of equinatoxin II: a study by UV-melting, DSC, far- and near-UV CD spectroscopy, and ANS fluorescence. AB - Thermal denaturation of equinatoxin II (EqTxII) in glycine buffer solutions (pH 1.1, 2.0, 3.0, and 3.5) and in triple distilled water (pH 5.5-6.0) was examined by differential scanning calorimetry, UV and CD spectroscopy and fluorescence emission spectroscopy of the added hydrophobic fluorescent probe ANS. At pH 5.5 6.0 and at temperatures below 60 degrees C, the protein exists in a native state characterized by a pronounced tertiary structure, a beta-rich secondary structure and a low degree of ANS-binding. At higher temperatures, it undergoes a two-state conformational transition, (delta H degree)VH = (delta H degree)DSC, into an unfolded state, which is characterized by a complete collapse of its tertiary structure and an incomplete denaturation of its secondary structure. At acidic pH, the EqTxII temperature-induced conformational transition appears at lower temperatures as non-two-state transition accompanied by the formation of an intermediate state which shows characteristics of molten globules, i.e., absence of defined tertiary structure, increase in alpha-rich secondary structure, and high affinity for ANS. At pH 2.0, the low-temperature initial state of EqTxII is already partially denatured; the tertiary structure is partially disrupted, and a pronounced inequality (delta H degree)VH > (delta H degree)DSC is observed. At pH value of 1.1 and below 60 degrees C, EqTxII exists in a stable acid-denatured compact state which shows all the characteristics of a molten globule, which even at 95 degrees C is not completely denatured. According to numerous studies on the pore forming toxins, such acid-denatured compact states may contribute to the protein's ability to penetrate into biological membranes. PMID- 9398153 TI - Kinetic hole burning, hole filling, and conformational relaxation in heme proteins: direct evidence for the functional significance of a hierarchy of dynamical processes. AB - Band III is a disorder and conformation-sensitive near-infrared (approximately 760 nm) charge transfer absorption band characteristic of equilibrium and nonequilibrium five coordinate ferrous high-spin hemes. The time evolution of this absorption band subsequent to photodissociation of six coordinate ferrous hemoglobin or myoglobin can provide detailed information regarding conformational relaxation, including the thermally driven fluctuations that result in the transition from inhomogeneous to homogeneous ligand rebinding kinetic. Such time resolved measurements over a range of temperatures are difficult due to long sample recovery times at cryogenic temperatures. A new restoring technique that allows for the rapid movement of a large optically accessible cryostat is used in combination with nanosecond time-resolved near-infrared absorption spectroscopy to generate band III as a function of time for the photoproducts of the carbon monoxide derivative of adult human hemoglobin (COHbA) and, to a more limited extent, horse myoglobin (COMb). The measurements are made over a wide range of temperatures extending from well below the solvent (75% glycerol:water) glass transition at approximately 180 K to ambient temperatures. Three temperature- and/or viscosity-dependent phenomena are observed. At the highest temperatures, only conformational relaxation is observed for the 75% glycerol sample. At very high viscosity (> or = 400 cp), conformational relaxation slows dramatically, and both kinetic hole burning followed by the filling in of the "hole" (dynamic hole filling) are observed. As the temperature is lowered, conformational relaxation slows and finally ceases. Kinetic hole burning and dynamic hole filling as well as additional broadening of band III are observed down to 140 K. The observation of kinetic hole burning (KHB) is indicative of the sample being inhomogeneous on the time scale of the ligand rebinding giving rise to KHB. The onset of hole filling is a direct manifestation of the thermal homogenization of the initial inhomogeneous distribution of conformational substates responsible for KHB. The observed dynamics are used to explain the inverse temperature effect associated with the non-Arrhenius slow down of geminate rebinding above approximately 180 K. The inverse temperature effect appears to arise not only from the onset of conformational relaxation but also from the increase in the rate on thermal averaging of the initial inhomogeneous distribution of conformational substates. PMID- 9398154 TI - Organophosphorus hydrolase is a remarkably stable enzyme that unfolds through a homodimeric intermediate. AB - Organophosphorus hydrolase (OPH, EC 8.1.3.1) is a homodimeric enzyme that catalyzes the hydrolysis of organophosphorus pesticides and nerve agents. We have analyzed the urea- and guanidinium chloride-induced equilibrium unfolding of OPH as monitored by far-ultraviolet circular dichroism and intrinsic tryptophan fluorescence. These spectral methods, which monitor primarily the disruption of protein secondary structure and tertiary structure, respectively, reveal biphasic unfolding transitions with evidence for an intermediate form of OPH. By investigating the protein concentration dependence of the unfolding curves, it is clear that the second transition involves dissociation of the monomeric polypeptide chains and that the intermediate is clearly dimeric. The dimeric intermediate form of OPH is devoid of enzymatic activity, yet clearly behaves as a partially folded, dimeric protein by gel filtration. Therefore, we propose an unfolding mechanism in which the native dimer converts to an inactive, well populated dimeric intermediate which finally dissociates and completely unfolds to individual monomeric polypeptides. The denaturant-induced unfolding data are described well by a three-state mechanism with delta G for the interconversion between the native homodimer (N2) and the inactive dimeric intermediate (I2) of 4.3 kcal/mol while the overall standard state stability of the native homodimer relative to the unfolded monomers (2U) is more than 40 kcal/mol. Thus, OPH is a remarkably stable protein that folds through an inactive, dimeric intermediate and will serve as a good model system for investigating the energetics of protein association and folding in a system where we can clearly resolve these two steps. PMID- 9398155 TI - Structure of pressure-assisted cold denatured lysozyme and comparison with lysozyme folding intermediates. AB - At high (> 3.5 kbar) pressures and low (< -10 degrees C) temperatures, hen egg white lysozyme denatures readily and reversibly. Amide hydrogen exchange methods were used to investigate the structure of the pressure-assisted cold-denatured state of lysozyme. Protection factors were obtained for 52 backbone amide protons. The extent of protection of many of these protons is markedly different from that in lysozyme denatured by high temperature, high urea concentration, or chemical modification; specifically, the protection factors are higher and are strongly correlated with elements of secondary structure present in the native state. Furthermore, the pattern of protection factors is similar to that observed in lysozyme during refolding from highly denatured states, particularly during the early stages (< 3.5 ms) of refolding [Gladwin, S. T., & Evans, P. A. (1996) Folding Des. 1, 407]. Previous data on cold-denatured ribonuclease A were reevaluated and compared to known folding intermediates [Houry, W. A. & Scheraga, H. A. (1996) Biochemistry 35, 11734; Udgaonkar, J. B., & Baldwin, R. L. (1990) Proc. Natl. Acad. Sci. U.S.A. 87, 8197] to further test the supposition that the pressure-assisted cold-denatured states of proteins resemble the early folding stages. PMID- 9398156 TI - Identification and localization of slow, natural, cooperative unfolding in the hematopoietic cell kinase SH3 domain by amide hydrogen exchange and mass spectrometry. AB - Protein unfolding on a fast time scale (milliseconds-minutes) has been widely reported, but slower unfolding events (10 min-hours) have received less attention. Amide hydrogen exchange (HX) and mass spectrometry (MS) were used to investigate the unfolding dynamics of the hematopoietic cell kinase (Hck) SH3 domain. Analysis of mass spectra after deuterium exchange into intact Hck SH3 indicates a cooperative unfolding event involving 24-61% of the domain and occurring with a half-life of approximately 20 min under physiological conditions. To identify the unfolding region, SH3 was incubated in D2O and proteolytically fragmented into peptides that were analyzed by mass spectrometry. Correlation of HX rates and isotope patterns reveals cooperative unfolding in several regions, including the C-terminal half of the RT-loop and a beta-sheet flanking the binding site. Binding of a prolyl-rich segment from the HIV Nef protein slows unfolding by a factor of 3. Further analysis yields a KD of 25 microM for the Nef peptide. These results demonstrate that an inherent flexibility in the SH3 domain may assist interconversion of the closed, intramolecularly ligated state and the open, active state of Src family kinases. Furthermore, this type of previously undetectable, slow unfolding process may provide the basis for new mechanisms in which kinetics of local unfolding combines with thermodynamics to regulate enzymatic activity. The combination of hydrogen exchange and mass spectrometry appears to be the only general method capable of examining these slow unfolding processes. PMID- 9398157 TI - VO2+(IV) complexes with pyruvate carboxylase: activation of oxaloacetate decarboxylation and EPR properties of enzyme-VO2+ complexes. AB - Chicken liver pyruvate carboxylase catalyzes a nonclassical ping-pong mechanism in which the carboxylation of biotin at subsite 1 of the active site is coupled to the biotin-dependent carboxylation of pyruvate at subsite 2. The functions of two divalent cation cofactors and at least one monovalent cation cofactor in catalysis are not well understood. The oxyvanadyl cation, VO2+ does not support phosphoryl transfer at the first subsite, and uncouples the decarboxylation of oxaloacetate at subsite 2 from the formation of ATP at subsite 1. Stimulation of this oxaloacetate decarboxylase activity in the presence of substrates and cofactors of the first subsite, including VO2+, VOADP-, Pi, and acetyl CoA, suggests that these cofactors and substrates induce the movement of carboxybiotin from the second subsite to the first subsite, where it is decarboxylated. VO2+ EPR has provided evidence for enzymic and nucleotide divalent cation binding sites within the first subsite. The EPR properties of enzyme bound VO2+ were altered by bicarbonate, suggesting that this substrate ligands directly to VO2+ at the enzymic metal site. Fluorescence quenching experiments suggest that a monovalent cation may interact with bicarbonate at the first subsite as well. The results of this study provide evidence that (i) the extrinsic metal ion cofactors interact with the substrates at the first subsite, and that (ii) divalent cations play a role in coupling catalysis at the two nonoverlapping subsites by inducing the decarboxylation of carboxybiotin at the first subsite. PMID- 9398158 TI - From ribonuclease A toward bovine seminal ribonuclease: a step by step thermodynamic analysis. AB - A proline, a leucine, and two cysteine residues, introduced at positions 19, 28, 31, and 32 of bovine pancreatic RNase A, i.e. the positions occupied by these residues in the subunit of bovine seminal RNase, the only dimeric RNase of the pancreatic-type superfamily, transform monomeric RNase A into a dimeric RNase, endowed with the same ability of BS-RNase of swapping its N-terminal segments. The thermodynamic consequences of the progressive introduction of these four residues into RNase A polypeptide chain have been studied by comparing the temperature- and urea-induced denaturation of three mutants of RNase A with that of a stable monomeric derivative of BS-RNase. The denaturation processes proved reversible for all proteins, and well represented by the two-state N<-->D transition model. The progressive introduction of the four residues into RNase A led to a gradual shift of the protein stability toward that characteristic of monomeric BS-RNase, which, in turn, is markedly less stable than RNase A with respect to both temperature- and urea-induced denaturation. On the other hand, the thermal stability of a dimeric active mutant of RNase A is found to approach that of wild-type seminal RNase. PMID- 9398159 TI - Dissociation of bacteriophage T4 DNA polymerase and its processivity clamp after completion of Okazaki fragment synthesis. AB - The mechanism of bacteriophage T4 DNA polymerase (gp43) and clamp (gp45) protein dissociation from the holoenzyme DNA complex was investigated under conditions simulating the environment encountered upon completion of an Okazaki fragment. Lagging strand DNA synthesis was approximated using a synthetic construct comprised of a doubly biotinylated, streptavidin-bound 62-mer DNA template, paired with complementary primers to generate an internal 12-base gap where the 5'-end primer contained either a 5'-OH (DNA primer) or a 5'-triphosphate (RNA primer) group. Rapid kinetic measurements revealed that upon encountering the blocking primer, the holoenzyme either dissociates from DNA (approximately 40%) or strand-displaces the blocking strand (approximately 60%). The two blocking oligonucleotides (DNA or RNA) induce a 30-50-fold increase in the rate of holoenzyme dissociation, with both polymerase and clamp proteins dissociating simultaneously. Inhibition of ATP hydrolysis by ATP-gamma-S did not have a measurable effect upon holoenzyme dissociation from DNA. The presence of gp32, the single-strand binding protein, caused a small (3-fold) increase in the rate constant for dissociation. PMID- 9398160 TI - Rapid deubiquitination of nucleosomal histones in human tumor cells caused by proteasome inhibitors and stress response inducers: effects on replication, transcription, translation, and the cellular stress response. AB - The proteasome inhibitors, lactacystin and N-acetyl-leucyl-leucyl-norlucinal, caused a rapid and near-complete loss of approximately 22-23-kDa ubiquitinated nucleoproteins, which we have identified as monoubiquitinated nucleosomal histones H2A and H2B by immunological and two-dimensional electrophoretic techniques. In human SKBr3 breast tumor cells, depletion of monoubiquitinated histones by the proteasome inhibitors coincided with the accumulation of high molecular weight ubiquitinated proteins in both nucleoprotein and cytosolic fractions and decreased unconjugated ubiquitin in the cytosol, without changes in the nonubiquitinated core histones. Unconjugated ubiquitin was not detected in isolated tumor cell nuclei. A similar loss in monoubiquitinated histones occurred in cells harboring a defective, temperature-sensitive mutation of the ubiquitin activating E1 enzyme, after these cells were elevated from 33 degrees C to the non-permissive temperature of 39 degrees C. DNA replication and RNA transcription were decreased by the proteasome inhibitors most strongly after 90% of the ubiquitin had been removed from ubiquitinated histones H2A and H2B, suggesting a relationship between the nucleosomal histone ubiquitin status and the processing of genetic information. Interestingly, although both proteasome inhibitors caused a generalized decrease in methionine incorporation into proteins, they strongly induced the synthesis of the hsp72 and hsp90 stress proteins. Finally, treating cells with heat-shock at 43 degrees C, with stress response-provoking chemicals or with several other proteasome inhibitors caused ubiquitinated proteins to accumulate, depleted free ubiquitin, and concomitantly decreased nucleosomal monoubiquitinated histones. These results suggest that deubiquitination of nucleosomal histones H2A and H2B may play a previously unrecognized role in the cellular stress response, as well as in the processing of chromatin, and emphasize the important role of the proteasome in cellular homeostasis. PMID- 9398161 TI - Role of glutamine-151 of human immunodeficiency virus type-1 reverse transcriptase in RNA-directed DNA synthesis. AB - Glutamine-151 of HIV-1 RT has been shown to be a catalytically important residue through the characterization of its mutant phenotype Glu151Ala (Sarafianos et al., 1995a). To further understand the role of this residue, we have extended this analysis to include polymerization on natural RNA template in addition to DNA template. We find that Q151A mutant exhibited a severe reduction in the polymerase activity without any significant effect on the affinity for dNTP substrate. Unlike DNA-directed reactions, the rate-limiting step for RNA-directed reactions does not appear to be either at the dNTP binding step or the chemical step. Analysis of the products formed on natural heteromeric HIV-genomic RNA template annealed with an 18-mer DNA primer with a sequence complementary to the primer binding site (PBS) has shown that addition of nucleotides is nonlinear with time since the enzyme appears to stall on the RNA template following the incorporation of the first nucleotide. The Q151A mutant was found to be nearly devoid of pyrophosphorolytic activity on a RNA-PBS template-primer. Similar properties have been previously reported for a mutant of R72 (R72A) of HIV-1 RT (Sarafianos et al., 1995b). However, R72 was implicated in stabilizing the transition state ternary complex before and after the phosphodiester bond formation (Kaushik et al., 1996; Sarafianos et al., 1995b). Our results with Q151A suggest that the side chain of Q151 may help stabilize the side chain of R72, and the loss of pyrophosphorolysis activity observed with the Q151 mutant may be the indirect manifestation of this stabilizing effect on R72. These observations point to the functional interdependence of residues Q151 and R72 in the polymerase function of the enzyme. An analysis of the 3D model structure of HIV-1 RT bound to DNA-DNA and RNA-DNA template-primer reveals that the guanidine hydrogen of R72 seems to stabilize Q151 by hydrogen bonding with its amide oxygen. A systematic conformational search of the side chain of Q151 also suggests a stable orientation where its specific interaction with the base of the RNA template may aid in stabilizing it. PMID- 9398162 TI - Phosphorylation of the acidic ribosomal P proteins in Saccharomyces cerevisiae: a reappraisal. AB - Previous reports had pointed to serines 62 and 71/79 as possible phosphorylation sites in the yeast acidic ribosomal proteins YP1 alpha and YP2 alpha, respectively. However, it has been found that mutation of these serine residues did not affect the phosphorylation level of the proteins. A detailed examination of the YP2 alpha tryptic digest from the in vivo labeled protein demonstrates the existence of a totally trypsin-insensitive site at lysine 88 that led to a misinterpretation of previous results. The unique YP2 alpha tryptic phosphopeptide obtained contains, in addition to serines 71 and 79, a serine at position 96 near the carboxyl end, which automatic Edman degradation confirmed as the phosphorylated residue. In addition, by using Staphyloccocus protease V8, it was possible to obtain phosphopeptides containing only serine 96, whose phosphorylation has likewise been confirmed by radioactive labeling as well as by chemical methods. A similar analysis of the other 12 kDa acidic proteins, YP1 alpha, YP1 beta, and YP2 beta, has shown the presence of equivalent phosphorylation sites in the four P proteins, which correspond to position 96 in proteins YP1 alpha, YP1 beta, and YP2 alpha and position 100 in YP2 beta. This conclusion has been confirmed by the fact that mutation of serine 96 in proteins YP1 alpha and YP2 alpha abolishes their capacity to be phosphorylated in vivo. The mutation of the phosphorylation site of the individual acidic proteins seems not to alter their interaction with the ribosome. However, it has been found that the level of phosphorylation of the stalk proteins has an effect on the response of the cells to some specific metabolic conditions, indicating that it may modulate the translation of specific proteins. PMID- 9398163 TI - ATF1 and CREB trans-activate a cell cycle regulated histone H4 gene at a distal nuclear matrix associated promoter element. AB - Proteins of the ATF/CREB class of transcription factors stimulate gene expression of several cell growth-related genes through protein kinase A-related cAMP response elements. The promoter activity of cell cycle regulated histone H4 genes is regulated by at least four principal cis-acting elements which mediate G1/S phase control and/or enhancement of transcription during the cell cycle. Using protein-DNA interaction assays we show that the H4 promoter contains two ATF/CREB recognition motifs which interact with CREB, ATF1, and ATF2 but not with ATF4/CREB2. One ATF/CRE motif is located in the distal promoter at the nuclear matrix-associated Site IV, and the second motif is present in the proximal promoter at Site I. Both ATF/CRE motifs overlap binding sequences for the multifunctional YY1 transcription factor, which has previously been shown to be nuclear matrix associated. Subnuclear fractionation reveals that there are two ATF1 isoforms which appear to differ with respect to DNA binding activity and partition selectively between nuclear matrix and nonmatrix compartments, consistent with the role of the nuclear matrix in regulating gene expression. Site-directed mutational studies demonstrate that Site I and Site IV together support ATF1- and CREB-induced trans-activation of the H4 promoter. Thus, our data establish that ATF/CREB factors functionally modulate histone H4 gene transcription at distal and proximal promoter elements. PMID- 9398164 TI - Effects of mutation of the conserved lysine-362 in cytochrome c oxidase from Rhodobacter sphaeroides. AB - We describe the effects of a mutation, K362M, of the conserved lysine in cytochrome c oxidase from Rhodobacter sphaeroides, a residue located in a putative proton channel that may convey substrate protons to the binuclear center. Spectra of the "as prepared", ferricyanide-oxidized, and dithionite reduced forms of the mutant protein confirm that the redox centers remain intact. Ligand binding kinetics of the ferricyanide-oxidized enzyme and of the dithionite reducible fraction are similar to those of the wild type, indicating that the K channel is not the major route for CO, cyanide, formate, or peroxide entry into the structure. Protonation of the lysine residue is not redox-linked to heme a or CuB as judged from the essentially unaltered midpoint potentials of these centers in the cyanide-ligated enzyme. A difficulty in electron transfer from heme a to the binuclear center is indicated by the slow and only partial reduction of heme a3 by dithionite or ferrocytochrome c and by the presence of some reduced heme a in the as prepared mutant enzyme and under steady-state conditions. Further characterization of the K362M enzyme in the steady state shows that up to one electron, but not two, can enter the binuclear center easily. It is this inability to form the two-electron-reduced, oxygen-reactive R state that prevents activity. A model is proposed where the K channel serves as a dielectric well of high dielectric strength and low proton conductivity, rather than as a pathway for proton entry to the binuclear center. The function of this structure would be to decrease the cost of introducing a transiently uncompensated charge into the binuclear center prior to formation of a stable, charge-compensated R-state. PMID- 9398165 TI - Formation of N delta-cyanoornithine from NG-hydroxy-L-arginine and hydrogen peroxide by neuronal nitric oxide synthase: implications for mechanism. AB - Neuronal nitric oxide synthase (nNOS) catalyzes the oxidation of NG-hydroxy-L arginine (NHA) by hydrogen peroxide. The amino acid products were characterized by high-performance liquid chromatography/mass spectrometry of the o phthalaldehyde/2-mercaptoethanol derivatives and identified as citrulline and N delta-cyanoornithine (CN-orn). The assignment of CN-orn was confirmed by independent chemical synthesis and comparison of the properties of the enzyme derived product with those of synthetic CN-orn. The inorganic products detected in the enzymatic reaction were NO2- and NO3-, presumably from oxidation of NO-. The reaction of H2O2 and NHA with nNOS was at least 10-fold slower than the reaction of NADPH, O2, and NHA (Vmax,app = 49 +/- 2 nmol min-1 mg-1 for the reactions with 10 microM added H4B). The reaction exhibited saturation kinetics with respect to hydrogen peroxide [K(m,app)(H2O2) = 10 +/- 1 mM for the reactions with 10 microM added H4B]. No H2O2-dependent reaction was observed with L arginine as the amino acid substrate. The different products for the NADPH- and H2O2-dependent transformations of NHA are of mechanistic significance in the NOS reaction. PMID- 9398166 TI - Yz. reduction kinetics in the absence of the manganese-stabilizing protein of photosystem II. AB - Decay of Signal IIvf of photosystem II (PSII), under repetitive flash conditions, was examined in whole cells of wild-type Synechocystis sp. PCC6803 and in cells of an engineered strain, delta psbO, which lacks the extrinsic 33 kDa manganese stabilizing protein (MSP). Previous polarographic analysis had shown that O2 release during the S3-->[S4]-->S0 transition of the catalytic cycle is significantly retarded in the delta psbO strain relative to the wild-type [Burnap et al. (1992) Biochemistry 31, 7404-7410]. The present experiments provide evidence that a parallel retardation in the rate of reduction of photooxidized Yz by the H2O oxidation complex is due to the absence of MSP. The half-time of the Signal IIvf component, corresponding to Yz. reduction during the S3-->[S4]-->S0 transition, was estimated to be 1.2 and 6.0 ms in the wild-type and delta psbO cells, respectively. PMID- 9398167 TI - Solution structure of the aminofluorene-intercalated conformer of the syn [AF]-C8 dG adduct opposite a--2 deletion site in the NarI hot spot sequence context. AB - This paper addresses structural issues related to the capacity of aminofluorene [AF] for frameshift mutations of the -2 type on C8 covalent adduct formation at the G3 site in the d(C-G1-G2-C-G3-C-C) NarI hot spot sequence. This problem has been approached from a combined NMR and relaxation matrix analysis computational structural study of the [AF]dG adduct in the d(C-G-G-C-[AF]G-C-C).d(G-G-C-C-G) sequence context at the 12/10-mer adduct level (designated [AF]dG.del(-2) 12/10 mer). The proton spectra of this system are of exceptional quality and are consistent with the formation of an AF-intercalated conformer with the modified guanine in a syn alignment displaced along with the 5'-flanking cytosine residue into the major groove. The solution structure has been determined by initially incorporating intramolecular and intermolecular proton-proton distances defined by lower and upper bound deduced from NOESY spectra as restraints in molecular mechanics computations in torsion angle space and subsequently refined through restrainted molecular dynamics calculations based on a NOE distance and intensity refinement protocol. Strikingly, the [AF]dG.del(-2) 12/10-mer duplex adopts only one of two potential AF-intercalation alignments for the [AF]dG adduct opposite the -2 deletion site in the NarI sequence context with the extrusion of the dC [AF]dG step favored completely over extrusion of the [AF]dG-dC step at the lesion site. This polarity establishes that the structural perturbation extends 5' rather than 3' to the [AF]dG lesion site in the adduct duplex. This structure of the [AF]dG adduct opposite a -2 deletion site shows distinct differences with conclusions reported on the alignment of the related acetylaminofluorene [AAF]dG adduct opposite a -2 deletion site in the identical NarI sequence context [Milhe, C., Fuchs, R. P. P., and Lefevre, J. F. (1996) Eur. J. Biochem. 235, 120-127]. In that study, qualitative NMR data without computational analysis were employed to conclude that the extrusion at the lesion site occurs at the [AAF]dG-dC step for the AAF-intercalated conformer of the adduct duplex. The structure of the [AF]dG adduct opposite a -2 deletion site determined in our group provides molecular insights into the architecture of extended slipped mutagenic intermediates involving aromatic amine intercalation and base-displaced syn modified guanines in AF and, by analogy, AAF-induced mutagenesis in the NarI hot spot sequence context. PMID- 9398168 TI - Solution structure of the aminofluorene-stacked conformer of the syn [AF]-C8-dG adduct positioned at a template-primer junction. AB - A solution structural study has been undertaken on the aminofluorene-C8-dG ([AF]dG) adduct located at a single strand-double strand d(A1-A2-C3-[AF]G4-C5-T6 A7-C8-C9-A10-T11-C12-C13).d (G14-G15-A16-T17-G18-G19-T20-A 21-G22) 13/9-mer junction (designated [AF]dG 13/9-mer) using proton-proton distance and intensity restraints derived from NMR data in combination with a computational protocol, which includes intensity refinement. This single strand-double strand junction models one arm of a replication fork composed of a 13-mer template strand, which contains the [AF]dG modification site, and a 9-mer primer strand, which has been elongated up to, but not including, the modified guanine. The NMR data establish that the duplex segment retains a minimally perturbed B-DNA conformation including Watson-Crick hydrogen-bonding at the junctional dC5.dG22 base pair. The NMR spectra are consistent with the guanine ring of the [AF]dG4 adduct adopting a syn glycosidic torsion angle and being displaced into the major groove with the adjacent dC3 residue displaced into the minor groove. Such a base displacement of the modified guanine is accompanied by stacking of one face of the fluorene ring of [AF]dG4 with the dC5.dG22 base pair, while the other face of the flourene ring is stacked with the purine ring of the nonadjacent dA2 residue in the intensity refined solution structures of the [AF]dG 13/9-mer. A comparison of structural features of the C8-[AF]dG adduct (this study) with those of the (+)-trans-anti-N2 [BP]dG adduct [Cosman et al. (1995) Biochemistry 34, 15334-15350] in the same 13/9-mer junctional sequence context has identified common features associated with the alignment of the modified guanine adducts at the template-primer junction. Thus, despite differences in the covalent linkage site for the C8 [AF]dG and (+)-trans-anti-N2-[BP]dG adducts, one face of the aromatic ring of the carcinogen stacks over the junctional base pair and in so doing displaces the modified guanine in a syn alignment into the major groove. These results lend credence to earlier proposals that such an adduct alignment may represent a common mutagenic conformer at a template-primer junction associated with a replication fork. PMID- 9398169 TI - Solution conformation of an intramolecular DNA triplex containing a nonnucleotide linker: comparison with the DNA duplex. AB - The solution properties of the parallel intramolecular DNA triplex d(GAGAGA-oct TCTCTC-oct-CTCTCT) (oct = -O-(CH2)8-O-PO2-O-(CH2)8-O-PO2-) and the duplex d(GAGAGA-oct-TCTCTC) have been examined by UV melting and high-resolution nuclear magnetic resonance spectroscopy (NMR). All nucleotides were primarily in the S conformation (i.e. near C2'-endo) in both the duplex and the triplex. However, the sugars of the Hoogsteen pyrimidine strand had a lower fraction of the S state than the Watson-Crick strands. Glycosidic torsion angles derived from nuclear Overhauser effect (NOE) build-up curves were found in the range -103 degrees to 133 degrees, with a clear alternation in magnitude along the GAGAGA strand in the triplex, whereas the glycosidic torsion angles were more similar in the duplex. Internucleotide NOEs were also consistent with an overall B-like geometry, rather than the A family. However, particularly in the Hoogsteen strand, some sequential NOE intensities were intermediate between those of the B and A forms. Distance and torsion constraints derived from NMR experiments were used to generate structures and were refined by restrained molecular dynamics. Extensive chemical shift differences between residues in the triplex and duplex were found for the purine strand, and there were remarkable differences in the pattern of shift differences for the A and G residues that correlated with differences in glycosidic torsion angles. Although there are differences in structure between the free duplex and that in the triplex, they are in important respects similar, indicating that only small conformational adjustments are needed to make parallel triple helices. PMID- 9398170 TI - Thermodynamic and kinetic basis of interfacial activation: resolution of binding and allosteric effects on pancreatic phospholipase A2 at zwitterionic interfaces. AB - A general kinetic model for catalysis by interfacial enzymes is developed. It couples the Michaelis-Menten catalytic turnover cycle at the interface with that in the aqueous phase through the distribution equilibria between the interface and the surrounding aqueous phase. Analysis under two limiting conditions fully describes the steady-state kinetics of hydrolysis and resolves the allosteric effects from apparent modes of interfacial activation in terms of the primary rate and equilibrium parameters for pig pancreatic phospholipase A2 (PLA2). One limit is observed in dispersions of anionic phospholipid vesicles, in which intervesicle exchange of enzyme, substrate, and hydrolysis products is absent and reaction occurs only on vesicles containing enzyme. A complete analysis at this highly processive limit, called kinetics in the scooting mode, has been published [Berg et al. (1991) Biochemistry 30, 7283]. Here is reported the analysis in the other limit, PLA2-catalyzed hydrolysis of zwitterionic micelles of short-chain phosphatidylcholines, at which substrate and products are in rapid exchange. Hydrolysis occurs either in bulk aqueous solution with phospholipid monomers or at the micellar interface. Above the critical micelle concentration (cmc), the hydrolysis rate shows a hyperbolic dependence on the bulk substrate concentration present as micelles. This dependence, characterized by the fitting parameters KMapp and VMapp, is analyzed in terms of the primary rate and equilibrium constants. The kinetic analysis is based on the assumption that the microscopic steady-state condition is satisfied because substrate replenishment in the micro environment of the enzyme is fast relative to the catalytic turnover time. Added NaCl and anionic interface increase the hydrolysis rate in zwitterionic micelles dramatically. The overall interfacial rate enhancement is attributed to three factors: (a) promotion of PLA2 binding by net anionic charge of the interface, (b) enhancement of substrate affinity of PLA2 at the interface (Ks* allostery), and (c) stimulation of the rate-limiting chemical step (kcat* allostery). PMID- 9398171 TI - Infrared spectroscopy of human apolipoprotein fragments in SDS/D2O: relative lipid-binding affinities and a novel amide I assignment. AB - Infrared absorption spectra are reported for six apolipoprotein fragments in SDS/D2O. Five of the peptides correspond to proposed lipid-binding domains of human apolipoproteins [apoC-I(7-24), apoC-I(35-53), apoA-II(18-30)+, apoA-I(166 185), apoE(267-289)], and the sixth is the de novo lipid associating peptide LAP 20. The amide I infrared absorption patterns are generally consistent with predominantly helical structures (as determined previously by NMR spectroscopy and distance geometry calculations) and further suggest that apoA-I(166-185) and apoE(267-289) are bound to SDS relatively weakly in comparison to the other four peptides. The latter conclusion is also supported by the temperature dependence of the infrared spectra, as increasing temperature promotes a distinct increase in random coil structure only for apoA-I(166-185) and apoE(267-289). In addition to features readily ascribed to helices, the infrared spectra of all the peptides show absorptions in the spectral region 1630-1635 cm-1 that is usually associated with beta-structure, a motif that is clearly absent from the NMR-derived structures. Parallel difficulties also arose in the analyses of the circular dichroism spectra. We suggest that both the low-frequency infrared absorptions and the ambiguities in interpreting the CD spectra may be due to unusual structures at the peptide C-termini, involving C=O groups that form hydrogen bonds simultaneously either with two solvent molecules or with donors from the backbone (NH) and the solvent (OH). Analogous absorptions may be a general feature of solvent-exposed helices, which suggests a need for caution in assigning amide I bands below 1640 cm-1. PMID- 9398172 TI - Dissecting the catalytic mechanism of staphylococcal lipases using carbamate substrates: chain length selectivity, interfacial activation, and cofactor dependence. AB - p-Nitrophenyl N-alkylcarbamates with different alkyl chains were used as substrates to determine separately the carbamylation and decarbamylation rates of the lipases from Staphylococcus hyicus and S. aureus. Both enzymes are reversibly inhibited by these compounds due to a rapid carbamylation of their active site serines followed by a slow decarbamylation. The carbamylation reaction is strongly pH-dependent and the pH profile suggests that an unprotonated histidine is required for this reaction. In contrast, the decarbamylation is pH-independent suggesting the presence of a hydrogen bond between the active site histidine and the carbamyl moiety. S. hyicus lipase preferably reacts with medium to long chain carbamates with an optimum for eight carbon atoms. In contrast, S. aureus lipase is highly specific for short chain carbamates. These results are in agreement with the respective substrate preferences of both lipases toward natural lipids. The decarbamylation rates of both enzymes hardly depend on the alkyl chain length, and from this it is concluded that chain length selectivity is expressed in the first step of catalysis. Both the carbamylation and decarbamylation reaction rates of S. hyicus lipase are enhanced in the presence of micelles, the activation effect being most pronounced in the first step. For the S. aureus lipase only a small influence of interfaces on both reaction steps was observed. These results are discussed in view of a possible role of a lid covering the active site. Kinetic experiments in the presence and absence of calcium strongly suggest that calcium ions are important for the structural stabilization of the unmodified as well as of the carbamylated enzymes. This structural function of calcium was supported by urea unfolding experiments, from which it appeared that for both enzymes the free energy for unfolding is significantly lower in the absence of calcium. In conclusion our results show that kinetic differences between both lipases reside in the acylation step, and that calcium is important for the structural stabilization of the unmodified, and moreover, the acylated enzymes. PMID- 9398173 TI - Kinetic and structural effects of mutations of the catalytic amino-terminal proline in 4-oxalocrotonate tautomerase. AB - The catalytic general base, Pro-1, of the enzyme 4-oxalocrotonate tautomerase has been mutated to Gly, Ala, Val, and Leu, residues with aliphatic side chains. The Val mutant was partially (55%) processed by removal of the amino-terminal methionine to yield P1V/M1P2V, while the Leu mutant was not processed and completely retained methionine (M1P2L). The M1P2L mutant lost 2300-fold in kcat with no change in Km, and the residual activity of the unresolvable P1V/M1P2V mixture could be explained by the summation of two activities, one equal to that of M1P2L and the other equal to that of the P1G mutant. The P1G and P1A mutants showed 76- and 58-fold decreases in kcat and much smaller decreases in Km of 4- and 2.8-fold, respectively. The dissociation constant of the substrate analog cis,cis-muconate decreased 1.7-fold in the P1G mutant as determined by NMR titration. 2D 1H-15N HSQC spectra and 3D 1H-15N NOESY HSQC spectra of the 15N labeled P1G mutant showed no structural differences from the wild-type enzyme except for small changes in backbone 15N and NH chemical shifts at the active site. Both the P1G and P1A mutants showed no change in overall conformation by circular dichroic spectroscopy. Both mutants and the wild-type enzyme generate the S-enantiomer of the product [5-2H]-2-oxo-3-hexenedioate with comparable stereoselectivities indicating a largely intact active site. The P1G and P1A mutants showed 10- and 4-fold decreases, respectively, in catalysis of exchange of the C3 proton of the substrate 2-oxo-1,6-hexanedioate, consistent with the lower basicities of Gly-1 and Ala-1 compared to Pro-1. The pH dependences of kcat/Km for the P1G and P1A mutants revealed pKa values of the general base of 5.3 and 5.9, respectively. NMR titration of the uniformly 15N-labeled P1G mutant showed the pKa of Gly-1 to be < or = 5.6, in agreement with the kinetic data. As with the wild-type enzyme, the active site environments on the P1G and P1A mutants lower the pKa of the general base by at least 2.5 units. It is concluded that the 2 order of magnitude decreases in kcat in the P1G and P1A mutants result from both a decrease in basicity and an increase in flexibility of the general base. The greater 10(3.4)-fold decrease in kcat found with the presence of an additional residue at the amino-terminus is ascribed to either the complete blockage or the drastically altered position of the general base. PMID- 9398174 TI - Heteronuclear NMR studies of the combined Src homology domains 2 and 3 of pp60 c Src: effects of phosphopeptide binding. AB - The results of heteronuclear NMR studies on the combined Src homology domains 2 and 3 (SH3-SH2) of pp60 c-Src are presented. Resonance assignments were obtained using heteronuclear triple-resonance experiments in conjunction with 15N separated nuclear Overhauser effect spectroscopy (NOESY) data. A modified three dimensional 13CO-15N-1H spectral correlation experiment [(HACA)CO(CA)-NH] with improved sensitivity is presented that provided additional sequential information and resolved several ambiguities. Chemical shifts and sequential- and medium range NOE cross peaks indicate that the structures of both the SH3 and SH2 portions of the polypeptide are very similar to those of the isolated SH3 and SH2 domains. Binding of a high-affinity phosphopeptide, EPQpYEEIPIYL, induces large chemical shift changes at several locations in the SH2 domain. Comparison with known results for peptide binding to SH2 domains shows that the residues displaying the largest effects are all involved in peptide binding or undergo significant conformational changes upon binding. However, subtle changes of both 1H and 15N chemical shifts are observed for residues within the SH3 domain and the connecting linker region, indicating possible cross-domain communication. PMID- 9398175 TI - Probing the conformation of NhaA, a Na+/H+ antiporter from Escherichia coli, with trypsin. AB - One of the most striking features of NhaA, an Escherichia coli Na+/H+ antiporter, is its extreme sensitivity to pH. The activity of NhaA increases 2000-fold between pH 6.5 and 8.5. In this work, we investigated whether the activation of NhaA by pH is accompanied by conformational changes which can be detected using trypsin as a probe. We have found that NhaA is susceptible to proteolytic digestion at the pH range where it is activated, suggesting that these two events may be related; at alkaline pH, the protein becomes active and adopts an "open" conformational state in which more domains are exposed to the enzyme. This idea was further supported by results from two mutants of NhaA in which His-225, a residue involved in pH sensing, has been replaced by either Arg or Asp. The mutant H225R is activated at more acidic pH values, while H225D at more alkaline pH. In accordance with the results described for the wild-type protein, H225R was susceptible to digestion by trypsin at the pH at which it undergoes main activation. NhaA has many potential tryptic cleavage sites. However, analysis of the tryptic digestion fragments suggests that at alkaline pH, the protein is exposed to cleavage mainly at hydrophilic loops 6, 7, and 8. Thus, upon activation, NhaA appears to undergo a change in conformation that is reflected in specific regions of the protein. PMID- 9398176 TI - An early step in Pseudomonas exotoxin action is removal of the terminal lysine residue, which allows binding to the KDEL receptor. AB - During the intoxication process, Pseudomonas exotoxin (PE) and immunotoxins containing PE internalize into the target cell and become processed into two fragments, and the carboxyl fragment translocates into the cytosol where it inactivates elongation factor 2. We have proposed that after internalization into cells the carboxyl-terminal fragment of PE (amino acids 280-613), which ends in REDLK, binds to the KDEL receptor (ERD2) which carries it to the endoplasmic reticulum, from which the PE fragment translocates to the cytosol. Earlier experiments showing that REDL but not REDLK binds to the KDEL receptor suggested that the terminal lysine is removed sometime during the intoxication process. To determine if and where this occurs, we exposed a peptide ending in REDLK to malignant cells in culture and found that binding to the KDEL receptor was restored. Restoration of receptor binding also occurred if a peptide or toxin ending in REDLK at its carboxyl terminus was incubated with plasma, indicating that the terminal lysine is removed prior to entry of the toxin into the cell. We conclude that plasma carboxypeptidase(s) cleave(s) the lysine residue from the carboxyl terminus of PE and PE-containing immunotoxins as an early and essential step in their cellular intoxication pathway. PMID- 9398177 TI - Proteolysis of the carboxyl-terminal GPI signal independent of GPI modification as a mechanism for selective protein secretion. AB - Variable amounts of soluble forms of a variety of glycosyl-phosphatidylinositol (GPI)-anchored proteins occur extracellularly, but the molecular mechanisms governing their release are not entirely clear. When the GPI-anchored folate receptor (FR) type beta was expressed transiently in human 293 fibroblasts, there was a roughly equal distribution of [3H]folic acid binding protein between the cell surface and the medium after 24 h over a wide range of expression levels of FR-beta. The difference in apparent molecular masses between the soluble FR-beta and the PI-PLC-treated membrane protein indicated that the former was not released from the membrane by the action of phospholipase. Brefeldin A inhibited the release of soluble FR-beta from both the transfected 293 cells and stable recombinant CHO (CHO-FR-beta) cells while pre-existing levels of cell surface FR were unaltered suggesting the absence of a precursor-product relationship between the membrane-associated FR-beta and the soluble protein in the medium. [35S]Cysteine pulse-chase analysis was consistent with this finding. Interchanging of carboxyl-terminal peptides between FR-beta and FR-alpha revealed that the nature of the processed signal for GPI modification was responsible for the quantitative membrane anchoring of FR-alpha and the production of soluble FR beta. When total cell lysates were analyzed by Western blot, a diffuse band of apparent 41 kDa and three additional sharp bands of apparent 35, 33, and 29.3 kDa were seen. The 41 kDa band was identified as the PI-PLC sensitive cell surface receptor. Several mutant constructs of FR-beta, in which the carboxyl-terminal signal for GPI modification was either disrupted or deleted only gave the three lower bands. The three sharp bands from the wild-type and the mutant forms of FR beta were identified as nonglycosylated (29.3 kDa) or glycosylated polypeptides in which the carboxyl-terminal peptide was at least partially proteolyzed without GPI modification. All of the mutations in the GPI signal resulted in the recovery of [3H]folic acid binding protein in the media which, similar to the wild-type FR recovered from the media, were converted to the 29.3 kDa band by N-glycanase. The results from this study indicate that a carboxyl-terminal peptide in FR-beta is efficiently proteolyzed intracellularly by a pathway that is independent of GPI signal recognition resulting in proper protein folding and secretion. Such carboxyl-terminal sequences could represent a simple adaptation for proteins whose physiologic functions reside both at the cell surface and in extracellular fluids, allowing their selective and tissue-specific release. PMID- 9398178 TI - Early photolysis intermediates of gecko and bovine artificial visual pigments. AB - Nanosecond laser photolysis measurements were conducted on digitonin extracts of artificial pigments prepared from the cone-type visual pigment, P521, of the Tokay gecko (Gekko gekko) retina. Artificial pigments were prepared by regeneration of bleached gecko photoreceptor membranes with 9-cis-retinal, 9-cis 14-methylretinal, or 9-cis-alpha-retinal. Absorbance difference spectra were recorded at a sequence of time delays from 30 ns to 60 microseconds following excitation with a pulse of 477-nm actinic light. Global analysis showed the kinetic data for all three artificial gecko pigments to be best fit by two exponential processes. These two-exponential decays correspond to similar decays observed after photolysis of P521 itself, with the first process being the decay of the equilibrated P521 Batho<-->P521 BSI mixture to P521 Lumi and the second process being the decay of P521 Lumi to P521 Meta I. In spite of its large blue shift relative to P521, iso-P521 displays a normal chloride depletion induced blue shift. Iso-P521's early intermediates up to Lumi were also blue-shifted, with the P521 Batho<-->P521 BSI equilibrated mixture being 15 nm blue-shifted and P521 Lumi being 8 nm blue-shifted relative to the intermediates formed after P521 photolysis. The blue shift associated with the iso-pigment is reduced or disappears entirely by P521 Meta I. Similar blue shifts were observed for the early intermediates observed after photolysis of bovine isorhodopsin, with the Lumi intermediate blue-shifted 5 nm compared to the Lumi intermediate formed after photolysis of bovine rhodopsin. These shifts indicate that a difference exists between the binding sites of 9- and 11-cis pigments which persists for microseconds at 20 degrees C. PMID- 9398179 TI - Role of the P6-P3' region of the serpin reactive loop in the formation and breakdown of the inhibitory complex. AB - Serpins have a large external peptide loop known as the reactive loop. Part of the reactive loop functions as the primary recognition site for target proteases; however, the complete role of the reactive loop in determining serpin specificity is unclear. In the current study, we investigated the reactive loop region that could potentially interact with the extended binding site of target proteases; the P6-P3' region. We utilized a reactive loop switching strategy to determine the extent to which the inhibitory activity of alpha-1-protease inhibitor (PI) against human neutrophil elastase (HNE) could be transferred to alpha-1 antichymotrypsin (ACT), a serpin that does not inhibit HNE. A series of ACT-PI chimeras were constructed in which segments of increasing length taken from the P6-P3' region of PI replaced the corresponding residues of ACT. The effectiveness of each chimera as an inhibitor of HNE was assessed by measuring (1) the rate of inhibitory complex formation and (2) the rate of complex breakdown (complex stability). Although all the ACT-PI chimeras were fully functional against chymotrypsin-like proteases, the series of chimeras showed no consistent progress toward the production of an inhibitor with the inhibitory properties of PI. The most rapid complex formation and most stable complexes were observed for chimeras with the P3-P1 residues of PI, whereas extending the replacement region to the P6 residue resulted in a considerable decrease in both inhibitory parameters. In order to study two additional features of the PI reactive loop that may play a role in the presentation of the P6-P3' region to HNE, we constructed variants that contained a P4' proline and deleted the P6'-P9' residues. Changes on the prime side appeared to have little effect on rates of inhibition or complex stability. Overall, even the most effective chimeras demonstrated an inhibition rate constant at least 60-fold less than that observed for PI inhibition of HNE and the most long lived chimera-HNE complexes broke down more rapidly than PI-HNE complexes. These results indicate that residues in the reactive loop region predicted to contact a specific target protease cannot fully transfer inhibitory activity from one serpin to another, suggesting that specific reactive loop serpin body and serpin body-protease body interactions play a significant role in determining serpin inhibitory activity against target proteases. PMID- 9398180 TI - Hydrogen bonding at the active site of delta 5-3-ketosteroid isomerase. AB - The solution secondary structure of the highly active Y55F/Y88F "Tyr-14-only" mutant of delta 5-3-ketosteroid isomerase complexed with 19-nortestosterone hemisuccinate has been shown to consist of three helices, a six-stranded mixed beta-sheet, and five turns. The steroid binds near the general acid, Tyr-14, on helix 1, near the general base, Asp-38, on the first strand of the beta-sheet, and on the hydrophobic face of the beta-sheet [Zhao, Q., Abeygunawardana, C., & Mildvan, A. S. (1997) Biochemistry 36, 3458-3472]. On this hydrophobic face, Asp 99 is the only polar residue. Free isomerase shows a deshielded exchangeable proton resonance at 13.1 ppm assigned to the N epsilon H of neutral His-100. Its fractionation factor (phi = 0.79) and slow exchange with solvent suggest it to be buried or involved in an H-bond. The binding of dihydroequilenin or estradiol to isomerase induces the appearance of two additional deshielded proton resonances, one at 18.2 ppm assigned to the gamma-carboxyl proton of Asp-99, and the other, at 11.6 ppm, assigned to the zeta-OH proton of Tyr-14. While mutation of Asp-99 to Ala results in the disappearance of only the resonance near 18 ppm [Wu, R. W., Ebrahemian, S., Zwrotny, M. E., Thornberg, L. D., Perez-Alverado, G. C., Brothers, P., Pollack, R. M., & Summers, M. F. (1997) Science 276, 415-418], both of these resonances disappear in mutants lacking Tyr-14, suggesting an H-bonded catalytic diad, Asp-99-COOH--Tyr14-OH--O-steroid enolate. The catalytic diad is further supported by NOEs from the beta 1 and beta 2 protons of Asp-99 to the epsilon protons of Tyr-14, and from the zeta-OH proton of Tyr-14 to the gamma carboxyl proton of Asp-99, indicating close proximity of these two residues, and by other data from the literature. A strong, low-barrier H-bond between Asp-99 and Tyr-14 is indicated by the 6.2 ppm deshielding, low fractionation factor (phi = 0.34) and slow exchange of the resonance at 18.2 ppm. A normal H-bond between Tyr-14 and the steroid is indicated by the 1.8 ppm deshielding, fractionation factor of 0.97 and the slow exchange of the resonance at 11.6 ppm. It is suggested that the 10(4.7)-fold contribution of Tyr-14 to catalysis is made possible by strong H-bonding from Asp-99 in the catalytic diad which strengthens general acid catalysis by Tyr-14. It is also noted that highly deshielded proton resonance on enzymes between 15 and 20 ppm, assigned to low-barrier H-bonds, generally involve carboxyl groups. PMID- 9398181 TI - Site-directed spin-labeling study of the structure and subunit interactions along a conserved sequence in the alpha-crystallin domain of heat-shock protein 27. Evidence of a conserved subunit interface. AB - Site-directed spin-labeling (SDSL) was used to investigate the secondary structure, solvent accessibility, and tertiary and quaternary interactions along the sequence located between residues 133 and 144 in the alpha-crystallin domain of human heat-shock protein 27 (HSP 27). The sequence is conserved among mammalian sHSP and shows similarity to the region of highest homology between alpha A- and alpha B-crystallins. Eleven sequential single cysteine mutants were prepared and reacted with a sulfhydryl-specific spin-label. The accessibilities of attached nitroxide side chains to a paramagnetic probe in the aqueous solution were determined. Spectral line shapes were analyzed in terms of side-chain mobility and spatial proximity to nearby nitroxides. The sequence-specific mobilities and accessibilities varied with a period of 2, consistent with the presence of a beta-strand along the sequence. At even sites, the nitroxide environment is highly ordered with virtually no accessibility to the hydrophilic probe, indicating that one face of the strand is buried. Furthermore, spin-spin interactions between nitroxides in the oligomeric structure strongly suggest that equivalent strands from different subunits are in close spatial proximity. These structural characteristics are remarkably similar to those of the equivalent sequence in alpha A-crystallin [Berengian, A. R., Bova, M. P., and Mchaourab, H. S. (1997) Biochemistry 36, 9951-9957]. In both proteins, a beta-strand spans the sequence and is located at a subunit interface, indicating that one set of interactions between subunits, and its associated symmetry, is conserved. This is the first report of sequence-specific structural similarity between alpha A crystallin and HSP 27 and the first identification of a conserved secondary structural element in the alpha-crystallin domain. PMID- 9398182 TI - Comparison of the hemolytic activity and solution structures of two snake venom cardiotoxin analogues which only differ in their N-terminal amino acid. AB - Cardiotoxin analogues IV (CTX IV) and II (CTX II) isolated from the venom of Taiwan Cobra (Naja naja atra) differ in their amino acid sequence by a single amino acid at the N-terminal end. Leucine at the N-terminal end in CTX II is replaced by arginine in CTX IV. CTX IV is an unique snake venom cardiotoxin as it is the only cardiotoxin isoform known so far which possesses a positively charged residue at the N-terminal amino acid. All other cardiotoxins have a hydrophobic amino acid (leucine or isoleucine) at their N-terminal end. The aim of the present study is to understand the effect(s) of the presence of a cationic residue on the structure and functional properties of cardiotoxin(s). Comparison of the hemolytic activities of CTX IV and CTX II shows that lytic activity of the former is at least twice as that shown by the latter. Comparison of the solution structures of CTX IV and CTX II using two-dimensional NMR spectroscopy and dynamical simulated annealing technique reveals that the backbone fold of both the toxin isoforms is almost similar. The secondary structural elements in these two cardiotoxin isoforms consist of long, triple-stranded, as well as short, double-stranded, antiparallel beta-sheets. Thermal denaturation experiments showed that the structure of CTX IV is more stable than that of CTX II. Critical analysis of the three-dimensional structures of CTX IV and CTX II reveals the presence of a "cationic" cluster comprising of positively charged residues on the concave side of the CTX IV molecule. Similar clusters consisting of positively charged residues are not found in CTX II. The differential erythrocyte lytic activities of these two cardiotoxins are attributed to the difference(s) in the distribution of the positively charged residues in their three-dimensional structures. PMID- 9398183 TI - Differences in active site gorge dimensions of cholinesterases revealed by binding of inhibitors to human butyrylcholinesterase. AB - Amino acid sequence alignments of cholinesterases revealed that 6 of 14 aromatic amino acid residues lining the active center gorge of acetylcholinesterase are replaced by aliphatic amino acid residues in butyrylcholinesterase. The Y337 (F330) in mammalian acetylcholinesterase, which is replaced by A328 in human butyrylcholinesterase, is implicated in the binding of ligands such as huperzine A, edrophonium, and acridines and one end of bisquaternary compounds such as BW284C51 and decamethonium. Y337 may sterically hinder the binding of phenothiazines such as ethopropazine, which contains a bulky exocyclic substitution. Inhibition studies of (-)-huperzine A with human butyrylcholinesterase mutants, where A328 (KI = 194.6 microM) was modified to either F (KI = 0.6 microM, as in Torpedo acetylcholinesterase) or Y (KI = 0.032 microM, as in mammalian acetylcholinesterase), confirmed previous observations made with acetylcholinesterase mutants that this residue is important for binding huperzine A. Inhibition studies of ethopropazine with butyrylcholinesterase mutants, where A328 (KI = 0.18 microM) was modified to either F (KI = 0.82 microM) or Y (KI = 0.28 microM), suggested that A328 was not solely responsible for the selectivity of ethopropazine. Volume calculations for the active site gorge showed that the poor inhibitory activity of ethopropazine toward acetylcholinesterase was due to the smaller dimension of the active site gorge which was unable to accommodate the bulky inhibitor molecule. The volume of the butyrylcholinesterase active site gorge is approximately 200 A3 larger than that of the acetylcholinesterase gorge, which allows the accommodation of ethopropazine in two different orientations as demonstrated by rigid-body refinement and molecular dynamics calculations. PMID- 9398184 TI - A diminished role for hydrogen bonds in antifreeze protein binding to ice. AB - The most abundant isoform (HPLC-6) of type I antifreeze protein (AFP1) in winter flounder is a 37-amino-acid-long, alanine-rich, alpha-helical peptide, containing four Thr spaced 11 amino acids apart. It is generally assumed that HPLC-6 binds ice through a hydrogen-bonding match between the Thr and neighboring Asx residues to oxygens atoms on the {2021} plane of the ice lattice. The result is a lowering of the nonequilibrium freezing point below the melting point (thermal hysteresis). HPLC-6, and two variants in which the central two Thr were replaced with either Ser or Val, were synthesized. The Ser variant was virtually inactive, while only a minor loss of activity was observed in the Val variant. CD, ultracentrifugation, and NMR studies indicated no significant structural changes or aggregation of the variants compared to HPLC-6. These results call into question the role of hydrogen bonds and suggest a much more significant role for entropic effects and van der Waals interactions in binding AFP to ice. PMID- 9398185 TI - NMR studies of the role of hydrogen bonding in the mechanism of triosephosphate isomerase. AB - Triosephosphate isomerase (TIM) catalyzes the reversible interconversion of dihydroxyacetone phosphate (DHAP) and glyceraldehyde-3-phosphate (GAP), with Glu 165 removing the pro-R proton from C1 of DHAP and neutral His-95 polarizing the carbonyl group of the substrate. TIM and its complexes with the reactive intermediate analogs, phosphoglycolic acid (PGA) and phosphoglycolohydroxamic acid (PGH), were studied by 1H NMR at 600 MHz and at low temperature (-4.8 degrees C). His-95 shows an N epsilon H resonance at 13.1 ppm which shifts to 13.3 ppm in the TIM-PGA complex and to 13.5 ppm in the TIM-PGH complex. In the TIM-PGH complex, His-95 N epsilon H shows a slow, pH-independent exchange rate with water (kex = 80 s-1 at 30 degrees C, Eact = 19 kcal/mol), which is 44-fold slower than that of an exposed histidine suggesting partial shielding from bulk solvent, and a fractionation factor phi = 0.71 +/- 0.02 consistent with its donation of a normal hydrogen bond. The formation of the TIM-PGH complex results in the appearance of several deshielded proton resonances, including one at 14.9 ppm and one at 10.9 ppm which overlaps with another resonance. The resonance at 14.9 ppm is absent and the resonance at 10.9 ppm is much weaker in the TIM complex of PGA, which lacks the hydroxamic acid (-NHOH) moiety. 15N-labeled PGH was synthesized and the NH proton of free [15N]PGH shows a single 1H-15N HMQC cross peak with delta (1H) = 10.3 ppm and delta (15N) = 168 ppm which shifts to delta (1H) = 10.9 ppm and delta (15N) = 174 ppm in the TIM complex of [15N]PGH. The 15N-1H coupling in the complex indicates covalent N-H bonding, and the deshielded delta (15N) indicates a significant contribution of the imidate resonance form of PGH. The 14.9 ppm resonance is assigned to the NOH proton of bound PGH. This resonance shows a pH-independent exchange rate with water (kex = 3900 s-1 at 30 degrees C, Eact = 8.9 kcal/mol) which may reflect the dissociation of the TIM-PGH complex, and meets the criteria for a low-barrier hydrogen bond on the basis of the significant downfield shift of 6.2 ppm from the NOH proton of the model compound acetohydroxamic acid, and a very low fractionation factor phi = 0.38 +/- 0.06. In the X-ray structure of the TIM-PGH complex [Davenport, R. C., Bash, P. A., Seaton, B. A., Karplus, M., Petsko, G. A., and Ringe, D. (1991) Biochemistry 30, 5821], the NOH proton of bound PGH is hydrogen bonded to Glu 165. A low-barrier hydrogen bond from PGH NOH to Glu-165 suggests a dual role for Glu-165 in catalysis of proton transfer not only between the C1 and C2 carbons but also between the O1 and O2 oxygens in the interconversion of DHAP and GAP in wild type TIM. Such a mechanism, together with the measured exchange rate of the His-95 N epsilon H proton with solvent protons can accommodate the classical measurements of tritium incorporation from DHAP into GAP. PMID- 9398186 TI - Absence of observable biotin-protein interactions in the 1.3S subunit of transcarboxylase: an NMR study. AB - Transcarboxylase (TC) is a biotin-containing enzyme catalyzing the transfer of a carboxyl group from methylmalonyl-CoA to pyruvate to form propionyl-CoA and oxalacetate. The transfer is achieved via carboxylated biotin bound to a 1.3S subunit within the multisubunit enzyme complex. The 1.3S subunit of TC is a 123 amino acid polypeptide, to which biotin is covalently attached at Lys 89. We have overexpressed 1.3S in Escherichia coli and characterized the biotinylated and apo forms by 1D- and 2D-NMR spectroscopy. To search for protein-biotin interactions, which could modulate the reactivity of the biotin ring on the 1.3S subunit, we have compared the chemical shifts, relaxation parameters, and NH exchange rates of the ureido ring protons of free and 1.3S-bound biotin. These properties are similar for both forms of the biotin. Further, NOE experiments on 1.3S revealed no detectable cross peaks between biotin and the protein. Consistent with these findings, the 2D NMR data for holo- and apo-1.3S are essentially identical indicating little or no changes in conformation between the two forms of the protein. The conclusion that strong protein-biotin interactions do not exist in 1.3S contrasts with the findings for the biotin carboxylase carrier protein from E. coli acetyl-CoA carboxylase, which reveal significant biotin-protein contacts [Athappilly, F. K., and Hendrickson, W. A. (1995) Structure 3, 1407-1419]. Further, the biotin NH1' exchange rates determined for 1.3S show that in the region of optimal activity for TC (pH 5.5-6.5) acid-catalyzed exchange predominates. In this pH range the base-catalyzed rate is too small (< 1 s-1) to account for the turnover rate of the enzyme. Thus, the means by which the N1' atom is activated for nucleophilic attack of the carboxyl group in methylmalonyl CoA does not appear to depend on interactions within the 1.3S subunit alone; rather activation must occur at the interfaces of the subunits in the holoenzyme. PMID- 9398187 TI - 13C- and 15N-labeled peptide substrates as mechanistic probes of oligosaccharyltransferase. AB - The carboxamide moiety that links the carbohydrate and protein moieties in N linked glycoproteins has been unambiguously determined to arise intact from asparagine by the use of chemically synthesized Bz-[4-13C, 15N]Asn-Leu-Thr-NH2 as an oligosaccharyltransferase (OST) substrate. Bz-[4-13C]Asn-Leu-Thr-NH2 was also synthesized and used to evaluate a proposed mechanism of OST catalysis similar to that of glutamine-dependent amidotransferases using 15NH4OAc as a potential external nucleophile. Analysis of NMR and MS spectra of the isotopically labeled peptides and the resulting biosynthesized glycopeptides indicates that free 15NH3 is not lost from the doubly labeled substrate during catalysis nor can exogenous 15NH3 intercept any of several postulated enzyme-bound species. These results indicate that OST-catalyzed glycosylation does not follow a mechanism involving the transient generation of exchangeable "NH3". Thus, in contrast to several glutamine-dependent amidotransferases, OST catalysis does not lead to transient scission of the asparagine beta-carboxamide C-N bond. Together with previously published results, these data argue against nucleophilic activation of the asparagine beta-carboxamide moiety being the underlying chemical mechanism for OST-catalyzed glycosylation of peptides. PMID- 9398188 TI - Proton nuclear magnetic resonance investigation of the [2Fe-2S](1-)-containing "Rieske-type" protein from Xanthobacter strain Py2. AB - Proton NMR spectra of the Rieske-type ferredoxin from Xanthobacter strain Py2 were recorded in both H2O and D2O buffered solutions at pH 7.2. Several well resolved hyperfine-shifted 1H NMR signals were observed in the 90 to -20 ppm chemical shift range. Comparison of spectra recorded in H2O and D2O buffered solutions indicated that the signals at -11.4 (L) and -15.5 (M) ppm were solvent exchangeable and thus were assigned to the two histidine N epsilon 2H protons. The remaining observed signals were assigned based upon chemical shift, T1 values, and one-dimensional nuclear Overhauser effect (nOe) saturation transfer experiments to either C beta H or C alpha H protons of cluster cysteinyl or histidine ligands. Upon oxidation of the [2Fe-2S] cluster, only two broad resonances were observed, indicating that the two Fe(III) ions are strongly antiferromagnetically coupled. In addition, the temperature dependence of each observed hyperfine-shifted signal in the reduced state was determined, providing information about the magnetic properties of the [2Fe-2S]1- cluster. Fits of the temperature data observed for each resonance to equations describing the hyperfine shift with their Boltzmann weighting factors provided a delta EL value of 185 +/- 26 cm-1 which, in turn, gives -2J as 124 cm-1. These data indicate that the two iron centers in the reduced [2Fe-2S] Rieske-type center are moderately antiferromagnetically coupled. The combination of these data with the available spectroscopic and crystallographic results for Rieske-type proteins has provided new insights into the role of the Rieske-type protein from Xanthobacter strain Py2 in alkene oxidation. PMID- 9398189 TI - Resonance Raman characterization of reaction centers in which bacteriochlorophyll replaces the photoactive bacteriopheophytin. AB - Qy-excitation resonance Raman (RR) spectra are reported for two mutant reactions centers (RCs) from Rhodobacter sphaeroides in which the photoactive bacteriopheophytin (BPhL) is replaced by a bacteriochlorophyll (BChl) molecule, designated by beta L. One mutation, (M)L214H, yields the pigment change via introduction of a histidine residue at position M214. The other mutation, (M)L214H/(L)-E104V, removes the putative hydrogen bond between beta L and the native glutamic acid residue at position L104. The vibrational signatures of the beta L cofactors of the mutants are compared with one another and with those of the accessory BChls (BChlL,M) in both beta-mutant and wild-type RCs. The spectroscopic data reveal the following: (1) The beta L cofactor is a five coordinate BChl molecule with a histidine axial ligand. The conformation of beta L and the strength of the Mg-histidine bond are very similar to that of BChlL,M. (2) The beta L cofactor is oriented in the protein pocket in a manner similar to that of BPhL of wild-type. (3) The beta L cofactor of the (M)L214H mutant forms a hydrogen bond with glutamic acid L104 via the C9-keto group of the macrocycle. The strength of this hydrogen bond is identical to that formed between this protein residue and the C9-keto group of BPhL in wild-type. (4) The hydrogen bonding interaction at the C9-keto site induces secondary cofactor-protein interactions which involve the C2a-acetyl and Cb-alkyl substituent groups. Collectively, the vibrational signatures of beta L indicate that its intrinsic physicochemical properties are very similar to those of BChlL. Consequently, the initial charge-separated intermediate in beta-type RCs is best characterized as a thermal/quantum mechanical admixture of P+ beta L- and P+ BChlL-(P is the primary electron donor), as originally proposed by Kirmaier et al. [(1995) J. Phys. Chem. 99, 8903-8909]. PMID- 9398190 TI - Structural coupling between the oxygen-evolving Mn cluster and a tyrosine residue in photosystem II as revealed by Fourier transform infrared spectroscopy. AB - The flash-induced Fourier transform infrared (FTIR) difference spectrum of the oxygen-evolving Mn cluster upon S1-to-S2 transition (S2/S1 spectrum) was measured using photosystem II (PS II) core complexes of Synechocystis 6803 in which tyrosine residues were specifically labeled with 13C at the ring-4 position. The double-difference spectrum between the unlabeled and labeled S2/S1 spectra showed that the bands at 1254 and 1521 cm-1 downshifted by 25 and 15 cm-1, respectively, upon ring-4-13C-Tyr labeling. This observation indicates that there is a tyrosine residue coupled to the Mn cluster, and the vibrational modes of this tyrosine are affected upon S2 formation. From a comparison of the above band positions and isotopic shifts in the S2/S1 spectrum with those of the FTIR spectra of tyrosine in aqueous solution at pH 0.6 (Tyr-OH) and pH 13.4 (Tyr-O-) and of the YD./YD FTIR difference spectrum, the 1254 and 1521 cm-1 bands were assigned to the CO stretching and ring CC stretching modes of tyrosine, respectively, and this tyrosine was suggested to be protonated in PS II. The observation that the effect of the S2 formation on the tyrosine bands appeared as a decrease in intensity with little frequency change could not be explained by a simple electrostatic effect by Mn oxidation, suggesting that the Mn cluster and a tyrosine are linked via chemical and/or hydrogen bonds and the structural changes of the Mn cluster are transmitted to the tyrosine through these bonds. On the basis of previous EPR studies that showed close proximity of YZ to the Mn cluster, YZ was proposed as the most probable candidate for the above tyrosine. This is the first demonstration of the structural coupling between YZ and the Mn cluster in an intact oxygen-evolving complex. This structural coupling may facilitate electron transfer from the Mn cluster to YZ. Our observation also provides an experimental support in favor of the proton or hydrogen atom abstraction model for the YZ function. PMID- 9398191 TI - Fourier transform infrared difference spectroscopy of photosystem II tyrosine D using site-directed mutagenesis and specific isotope labeling. AB - Tyrosine D (TyrD), a side path electron carrier of photosystem II (PS II), has been studied by light-induced Fourier transform infrared (FTIR) difference spectroscopy in PS II core complexes of Synechocystis sp. PCC 6803 using the experimental conditions previously optimized to generate the pure TyrD./TyrD FTIR difference spectrum in PS II-enriched membranes of spinach [Hienerwadel, R., Boussac, A., Breton, J., and Berthomieu, C. (1996) Biochemistry 35, 115447 115460]. IR modes of TyrD and TyrD. have been identified by specific 2H- or 13C labeling of the tyrosine side chains. The v8a(CC) and v19(CC) IR modes of TyrD are identified at 1615 and 1513-1510 cm-1, respectively. These frequencies show that TyrD is protonated. Comparison of isotope-sensitive signals in situ with those of the model compound p-methylphenol dissolved in different solvents leads to the assignment of the v7'a(CO) and delta(COH) modes of TyrD at 1275 and 1250 cm-1, respectively. It is shown that these modes and in particular the delta(COH) IR mode are very sensitive to the formation of hydrogen-bonded complexes with amide C=O or with imidazole nitrogen atoms. The frequencies observed in situ show that TyrD is hydrogen-bonded to the imidazole ring of a neutral histidine. For the radical TyrD., isotope-sensitive IR modes are identified at 1532 and 1503 cm 1. The signal at 1503 cm-1 is assigned to the v(CO) mode of TyrD. since it is sensitive to 13C-labeling at the ring carbon involved in the C4-O bond. The perturbation of TyrD and TyrD. IR modes upon site-directed replacement of D2 His189 by Gln confirms that a hydrogen bond exists between both TyrD and TyrD. and D2-His189. In the D2-His189Gln mutant, the v7'a(CO) mode of TyrD at 1267 cm-1 and the delta(COH) mode at approximately 1228 cm-1 show that a hydrogen bond is formed between TyrD and an amide carbonyl, probably that of the D2-Gln189 side chain. Electron nuclear double resonance (ENDOR) measurements have shown that TyrD. is hydrogen-bonded in the wild type but not in the mutant [Tang, X.-S., Chrisholm, D. A., Dismukes, G. C., Brudwig, G. W., and Diner, B. A. (1993) Biochemistry 32, 13742-13748]. The v(CO) mode of TyrD. at 1497 cm-1 is downshifted by 6 cm-1 compared to WT PS II, indicating that hydrogen bonding induces a frequency upshift of the v(CO) IR mode of Tyr.. IR signals from the Gln side chain v(C=O) mode are proposed to contribute at 1659 and 1692 cm-1 in the TyrD and TyrD. states, respectively. These frequencies are consistent with the rupture of a hydrogen bond upon TyrD. formation in the mutant. The frequency of the v(CO) mode of TyrD., observed at 1503 cm-1 for WT PS II, is intermediate between that observed at 1497 cm-1 in the D2-His189Gln mutant and at 1513 cm-1 for Tyr. formed by UV irradiation in borate buffer, suggesting weaker or fewer hydrogen bonds for TyrD. in PS II than in solution. The role of D2-His189 in proton uptake upon TyrD. formation is also investigated. PMID- 9398192 TI - Surface of cytochrome c: infrared spectroscopy of carboxyl groups. AB - The carboxylate groups of organic acids give strong absorption in the infrared between approximately 1550 and 1650 cm-1. For acetate and chloroacetate derivatives, the infrared (IR) frequency of the carboxylate antisymmetric stretching mode (v(a)OCO) is related to the square root of the pK of the acid, with a shift of approximately 20 cm-1 to higher frequency for a pK drop in the range 5-3. It follows that v(a)OCO may respond to conditions on the protein surface. In this paper, the IR amide I' and carboxylate absorptions of cytochrome c from horse, yeast, and tuna are compared with model compounds such as Val-Glu and microperoxidase-11, the 11 amino acid fragment of horse cytochrome c containing the covalently bound heme. For microperoxidase-11, the contribution from all four carboxylates can be accounted for and the 1567 cm-1 absorption is assigned to the heme propionates. For the proteins, the carboxylate absorption band is inhomogeneous, i.e., there is a distribution of frequencies. Both the amide I' and carboxylate bands are sensitive to protein conformation as shown by their different pH, salt, and redox dependence. PMID- 9398193 TI - A rationale for the absolute conservation of Asn70 and Pro71 in mitochondrial cytochromes c suggested by protein engineering. AB - The absolutely conserved residues Asn70 and Pro71 of mitochondrial cytochrome c have been targeted for protein engineering by semisynthesis. Neither residue has even been implicated in mechanistic schemes, and we reasoned that the conservation of this dipeptide was to fulfill a crucial structural role. Semisynthesis was through condensation by autocatalytic fragment religation of natural fragment 1-65 (H) of the horse protein and synthetic 39-residue peptides containing noncoded amino acids prepared by solid-phase methods. High yields of the purified analogs, homoserine70 and norvaline71 cytochromes c, were obtained. Functional tests revealed minor destabilization of the Hse70-containing structure, with little adverse effect in in vitro assays, but [Nva71] cytochrome c was essentially devoid of activity in these systems. This appeared to be a consequence of a shift, more pronounced than any yet reported, in the conformational equilibrium between the active state III conformer and the inactive, 'alkaline' state IV. The results support our view that this dipeptide is optimal for, and rigidifies, the right-angle bend between two alpha-helices, thus determining the conformation of the 70s loop that terminates in the sixth ligand Met80, and 'forcing' the coordination of iron by thioether sulfur in the presence of the adjacent more avid amine ligands of state IV. Not only is [Nva71] cytochrome c inactive at pH 7, but it also proves to be an extremely potent inhibitor of electron transfer by native state III, thus providing the rationale for the evolutionary conservation of a high pK for the ligand exchange reaction. PMID- 9398194 TI - Kinetics of ferric cytochrome P450 reduction by NADPH-cytochrome P450 reductase: rapid reduction in the absence of substrate and variations among cytochrome P450 systems. AB - The reduction of ferric cytochrome P450 (P450) to ferrous is the first chemical step in almost all P450 reactions, and many characteristics of this step have been reported. Reduction kinetics of rabbit and human P450s were measured in a variety of systems. As reported earlier, P450 reduction is biphasic in microsomes and some purified P450 systems. However, this is not an inherent property of P450s, and some low- and high-spin iron P450s were reduced with single exponential kinetics. Contrary to a generalized view, the presence of substrate is not necessary for rapid reduction of all P450s. Also, low-spin heme can be reduced as rapidly as high-spin in several P450s. P450s varied considerably in their reduction behavior, and even a single P450 showed remarkably different reduction kinetics when placed in various environments. P450 3A4 reduction was examined in liver microsomes, a reconstituted system, a fusion protein in which it was linked to NADPH-P450 reductase, and baculovirus and bacterial membranes in which P450 3A4 and NADPH-P450 reductase were coexpressed; the systems differed considerably in terms of the need for the substrate testosterone and cytochrome b5 (b5) for reduction and as to whether reduction was rate-limiting in the overall catalytic cycle. When b5 was included in reconstituted systems, it reduction kinetics were linked with those of some P450s. This behavior could be simulated in kinetic models in which electrons flowed from the ferrous P450.CO complex to oxidized b5. Overall, the kinetics of ferric P450 reduction cannot be generalized among different P450s in various systems, and concepts regarding influence of substrate, reaction sequence, and a rate-limiting step are not very universal. PMID- 9398195 TI - Identification of a critical phenylalanine residue in horseradish peroxidase, Phe179, by site-directed mutagenesis and 1H-NMR: implications for complex formation with aromatic donor molecules. AB - The functional and structural significance of Phe179 of horseradish peroxidase isoenzyme C (HRP C) has been investigated by site-directed mutagenesis. This residue is located in a structurally variable insertion between helices F and G, a motif unique to peroxidases of higher plants. Results obtained for three recombinant enzymes, with Phe179 substituted by Ala, His, or Ser, provide the first demonstration of the importance of this side chain for the binding of aromatic donor molecules. Experimental parameters for direct comparison with the wild-type enzyme were obtained by extensive solution state characterization using both optical and 1H-NMR spectroscopy. Significant chemical shift variations for resonances associated with the exposed heme edge, notably heme methyl C18H3 and heme propionate C17(1)H2, were recorded in NMR spectra of both the resting and cyanide-ligated states of the three Phe179 mutants. Furthermore, comparison of NOE connectivities in NOESY spectra of cyanide-ligated wild-type and mutant enzymes enabled the elusive assignment of the aromatic side chain in close proximity to heme methyl C18H3 to be made to Phe179. Replacement of Phe179 by Ala resulted in an 80-fold decrease in the binding affinity of the cyanide-ligated enzyme for benzhydroxamic acid, with a Kd value similar to that determined for cyanide-ligated HRP A2 (an acidic isoenzyme with valine at position 179). The binding affinity of Phe179-->Ser was similarly decreased, while that of Phe179- >His was partially restored relative to wild-type HRP C. Cyanide-ligated Phe179- >His HRP C exhibited a unique pH-dependent spectral transition associated with a pKa value of 6.5 +/- 0.2, assigned to the His179 side chain. Two closely related enzyme forms exhibiting different affinities for benzhydroxamic acid were observed at neutral pH and above, indicating that the protonation state of His179 gave rise to microheterogeneity in the aromatic donor molecule binding site. PMID- 9398196 TI - Role of methionine-239, an amino acid residue in the mobile-loop region of the NADH-binding domain (domain I) of proton-translocating transhydrogenase. AB - Transhydrogenase couples the transfer of hydride equivalents between NAD(H) and NADP(H) to proton translocation across a membrane. The one-dimensional proton NMR spectrum of the recombinant NAD(H)-binding domain (domain I) of transhydrogenase from Rhodospirillum rubrum reveals well-defined resonances, several of which arise from a mobile loop at the protein surface. Four have been assigned to Met residues (MetA-MetD). Substitution of Met239 with either Ile (dI.M239I) or Phe (dI.M239F) resulted in loss of MetA from the NMR spectrum. Broadening and shifting of the mobile loop resonances consequent on NAD(H) binding indicate that the loop closes down on the protein surface. More NAD(H) had to be added to mutant domain I than to wild type to give comparable resonance broadening. The Kd of domain I for NADH, measured by equilibrium dialysis, was increased about three fold by the Met239 mutations. Mutant and wild-type domain I were reconstituted with domain I-depleted membranes from R. rubrum, and with recombinant domain III of transhydrogenase. With membranes, the Km for acetylpyridine adenine dinucleotide during reverse transhydrogenation was 5x and > 6x greater in dI.M239I and dI.M239F, respectively, than in wild-type. Cyclic transhydrogenation (in membranes and the recombinant system) was substantially more inhibited (70% in dI.M239I, and 84% in dI.M239F) than either forward or reverse transhydrogenation. The docking affinities of dI.M239I and dI.M239F to the depleted membranes were similar to those of wild-type. It is concluded that Met239 is MetA in the mobile loop of domain I, and that in proteins with amino acid substitutions at this position, the binding affinity of NAD(H) is decreased, and the hydride transfer step is inhibited. PMID- 9398197 TI - A three-domain iron-sulfur flavoprotein obtained through gene fusion of ferredoxin and ferredoxin-NADP+ reductase from spinach leaves. AB - Ferredoxin and ferredoxin-NADP+ reductase are the two last partners of the photosynthetic electron-transfer chain, responsible for the final reduction of NADP+ to NADPH. Herein, we report the engineering and characterization of a novel protein molecule in which the electron-carrier protein (ferredoxin I) and the reductase (a flavoprotein) were covalently linked in a single polypeptide chain by gene fusion. The gene was obtained by joining the cDNAs encoding the respective proteins and subsequently by deleting the intervening sequence between them by site-directed mutagenesis. No extra amino acid residues were introduced between the C-terminus of ferredoxin I and the N-terminus of the flavoenzyme. The chimera was purified to homogeneity and characterized. The M(r) of the chimera apoprotein was 45,800 as determined by mass spectrometry, in agreement with the expected value of 45,846. Both flavin and iron-sulfur cluster were in 1:1 ratio with respect to the apoprotein. The chimera was found active as a diaphorase and, more interestingly, highly efficient as a cytochrome c reductase, without need for free ferredoxin addition in the assay medium. Several lines of evidence indicate that the ferredoxin and the reductase in the chimera assume a configuration quite similar to that in the dissociable physiological complex. Thus, the fusion protein could be a useful tool for studying the mechanism of protein-protein recognition and electron transfer in the ferredoxin-ferredoxin NADP+ reductase system. PMID- 9398198 TI - RNA secondary structure switching during DNA synthesis catalyzed by HIV-1 reverse transcriptase. AB - Changes in RNA secondary structure have been found to play important roles in translational regulation, protein synthesis, and mRNA splicing. In studies utilizing a 66 nucleotide RNA template with a stable hairpin structure, we have examined the effects of RNA secondary structure on HIV-1 reverse transcriptase activity. We identify several pause sites in the stem of the hairpin and show that these pause sites are correlated with the free energy of melting the next base pair in the stem. We also identify a pause site appearing in the loop of the hairpin and show that this is due to the rapid formation of a new hairpin structure occurring during the progress of DNA polymerization through the hairpin. The rapid change in RNA secondary structure to form the new hairpin selectively destabilizes the major hairpin and thereby accelerates the rate at which reverse transcriptase reads through RNA secondary structure. PMID- 9398199 TI - Decreased Stability of Transforming Growth Factor beta Type II Receptor mRNA in RER+ Human Colon Carcinoma Cells AB - Transforming growth factor beta (TGF-beta) is a potent inhibitor of cell growth and tumor progression. Previous work has shown that loss of functional TGF-beta type II receptor (RII) due to a frameshift mutation in the 5' half of the RII gene leads to TGF-beta resistance in a highly progressed, RER+ human colon carcinoma cell line designated HCT116. Expression of this mutated RII gene was highly repressed in RER+ cell lines such as HCT116 and RKO, as analyzed by RNase protection assays. Nuclear run-on and RII promoter-reporter (CAT) assays showed that the transcriptional levels of the RII gene in these RER+ cells were not reduced, compared to RII-expressing cells. However, the half-lives of the RII mRNA, as analyzed by RNase protection assays following actinomycin D treatment, were significantly decreased. This suggested that the decreased expression of the RII gene mutant was due to decreased mRNA stability. Furthermore, RII mRNA from HCT116 transfected with wild-type RII had a longer half-life than the endogenous mutated RII mRNA. A dominant negative RII mutant, which encodes a similarly truncated RII protein as HCT116 but lacks the extensive 3' untranslated region of RII mRNA, gave the same half-life as endogenous wild-type RII mRNA. We conclude that the frameshift mutation which results in a premature stop codon in the 5' half of the mRNA transcript accounts for the reduced RII mRNA levels in RER+ cells. PMID- 9398200 TI - Specificity of an Escherichia coli RNA polymerase-associated NTPase. AB - Standard preparations of Escherichia coli RNA polymerase harbor a 70 kDa protein with NTPase (beta-gamma cleavage) activity that is not a recognized polymerase subunit. The NTPase activity of this component, before and after separation from the polymerase, is strongly dependent on the presence of DNA; single-stranded polydeoxynucleotides are more effective than double-stranded. ATP and GTP are cleaved, the latter much less readily. The NTPase as it occurs with the polymerase displays cleavage preference for NTPs that are not complementary to the DNA, a fact that has led to proposals for involvement of the NTPase in transcriptional error prevention [Volloch, V. Z., Rits, L. & Tumerman, L. (1979) Nucleic Acids Res. 6, 1535-1546; Libby, R. T., Nelson, J. L., Calvo, J. M., & Gallant, J. A. (1989) EMBO J. 8, 3253-3158]. We find, however, that the lesser cleavage in the presence of complementary DNA results from competition for the NTP between the processes of incorporation by the polymerase and of cleavage by the NTPase, operating on the same substrate pool. The greater cleavage with noncomplementary DNA occurs because of the lack of incorporation by the polymerase, which then does not compete with the NTPase for the substrate pool. Thus, these findings indicate that the cleavage preference of the NTPase for noncomplementary NTPs is not part of a mechanism for error prevention during transcription. PMID- 9398201 TI - A unique transcription factor for the A alpha fibrinogen gene is related to the mitochondrial single-stranded DNA binding protein P16. AB - Although stimulation of hepatic cells with interleukin-6 induces the expression of fibrinogen, the molecular basis for this regulation remains largely uncharacterized. A recent examination of the A alpha fibrinogen gene promoter identified a protein, termed the A alpha-core protein, that bound constitutively to the IL-6 response element [Liu, Z. & Fuller, G. M. (1995) J. Biol. Chem. 270, 7580-7586]. This current study provides further characterization of this regulatory protein. The data presented show the following: (i) The A alpha-core protein has a similar molecular weight and identical N-terminal sequence to that of the mitochondrial single-stranded DNA binding protein P16. (ii) The A alpha core protein and P16 have similar characteristics in terms of DNA binding preference and antigenic properties. (iii) Overexpression of P16 gene in the hepatoma cell lines Hep G2 and Hep 3B enhances the IL-6-induced expression of A alpha fibrinogen. These results demonstrate that the A alpha-core protein is closely related to P16 and involved in the IL-6-regulated transcription of A alpha fibrinogen. PMID- 9398202 TI - Binding of tsHMG, a mouse testis-specific HMG-domain protein, to cisplatin-DNA adducts. AB - The anticancer drug cisplatin is particularly effective against testicular tumors. Although the clinical consequences of cisplatin chemotherapy are well known, the precise mechanism of action remains elusive. Specific recognition of cisplatin-damaged DNA by a class of proteins containing the high-mobility group (HMG) domain DNA-binding motif could play a role in mediating the cytotoxicity of the drug. This study presents a quantitative investigation of binding of the murine testis-specific high-mobility group protein tsHMG to DNA modified by cisplatin. The binding affinity and specificity of this protein to a site specific 1,2-d(GpG) cisplatin-DNA intrastrand cross-link in a 20 bp probe were determined. A value for the apparent dissociation constant, Kd(app), of 24 +/- 5 nM was obtained by gel mobility shift assays. Binding competition assays with the corresponding unmodified 20 bp probe gave a ratio (rho) of nonspecific to specific Kd(app) values of 230. A polypeptide containing tsHMG domain A (residues 1-82) was expressed and purified to homogeneity. This domain alone was sufficient for specific recognition of cisplatin-modified DNA with a Kd(app) of 300 +/- 50 nM and a rho of 20, a comparatively high discrimination factor. DNase I interference analysis of the adduct-containing strand revealed that tsHMG binding extends over 14 nucleotides, centered around the platinated bases. The domain A polypeptide protection pattern covers a slightly smaller area of 13 nucleotides. The binding affinity and specificity of tsHMG for cisplatin-modified DNA are exceptional compared to those of other HMG-domain proteins studied previously. The possible relevance of these findings to the mechanism of action of cisplatin is discussed. PMID- 9398203 TI - Modulation of Cm/T, G/A, and G/T triplex stability by conjugate groups in the presence and absence of KCl. AB - Apparent equilibrium association constants were determined by gel mobility shift analysis for triple strand formation between a duplex target containing a 21 base long A-rich homopurine run and several end-modified C(m)/T (pyrimidine motif; C(m) = 5-methylcytosine), G/A (purine motif), and G/T (purine-pyrimidine motif) triplex-forming oligonucleotides (TFOs). Incubations were carried out for 24 h at 37 degrees C in 20 mM HEPES, pH 7.2, 10 mM MgCl2, and 1 mM spermine. The purine motif triplex was the most stable (Ka = 6.2 x 10(8) M-1) even though the TFO self associated as a linear duplex. Conjugation of a terminal hexanol or cholesterol group to the G/A-containing TFO reduced triplex stability by 1.6- or 13-fold, whereas an aminohexyl group or intercalating agent (acridine or psoralen) increased triplex stability by 1.3- or 13-fold. These end groups produced similar effects in C(m)/T and G/T triplexes, although the magnitude of the effect sometimes differed. Addition of 140 mM KCl to mimic physiological conditions decreased stability of the G/A triplex by 1900-fold, making it less stable than the C(m)/T triplex. The inhibitory effect of KCl on G/A triplex formation could be partially compensated for by conjugating the TFO to an intercalating agent (30 350-fold stabilization) or by adding the triplex selective intercalator coralyne (1000-fold stabilization). Although the G/T triplex responded similarly to these agents, the stability of the C(m)/T triplex was unaffected by the presence of coralyne and was only enhanced 1.4-2.8-fold when the TFO was linked to an intercalating agent. In physiological buffer supplemented with 40 microM coralyne, the G/A triplex (Ka = 3.0 x 10(8) M-1) was more stable than the C(m)/T and G/T triplexes by factors of 300 and 12, respectively. PMID- 9398204 TI - Structural and mechanistic studies on chloroplast translational initiation factor 3 from Euglena gracilis. AB - Chloroplast translational initiation factor 3 (IF3chl) from Euglena gracilis contains a central region (homology domain) that is homologous to prokaryotic IF3. The homology domain is preceded by a long NH2-terminal extension (head), and followed by a 64 amino acid COOH-terminal extension (tail). Sequences in these extensions reduce the activity of the homology domain. To gain insight into these effects, a possible three-dimensional structure for the homology region of IF3chl has been modeled using the X-ray coordinates from the N- and C-domains of Bacillus stearothermophilus IF3. In B. stearothermophilus IF3, these two compact domains are thought to fold independently and are separated by a helical lysine rich linker. The modeled structure suggests that IF3chl has a similar overall fold although some subtle differences are predicted to occur. Both the head and tail regions of IF3chl are oriented toward the linker region in the homology domain where they may potentially interfere with its function. Circular dichroism spectropolarimetry (CD) indicates that the lysine-rich linker region in IF3chl is not in a helical conformation and is probably a random coil. CD analysis indicates that a portion of the tail region of IF3chl is helical and that the tail has a direct interaction with the linker region in the homology domain. Site directed mutagenesis of the linker indicates that two conserved lysine residues are important for the function of IF3chl and play a role in the binding of IF3chl to the 30S ribosomal subunit. Mutation of these residues affects the interaction of the homology domain with the tail. PMID- 9398205 TI - Triple helix formation and homologous strand exchange in pyrene-labeled oligonucleotides. AB - The orientations of the symmetrical third strands (G3A4G3) and (G3T4G3) within the triplexes (C3T4C3) - (G3A4G3) x (G3A4G3) and (C3T4C3) - (G3A4G3) x (G3T4G3) were investigated by fluorescence spectroscopy and thermal denaturation using pyrene-labeled oligodeoxynucleotides. In the two triplex structures, both parallel and antiparallel orientations of the third strand with respects to the purine Watson-Crick one were identified by means of pyrene excimer formation. The pyrene labels do not modify the melting temperature of the (C3T4C3) - (G3A4G3) x (G3T4G3) triplex but somewhat stabilize the corresponding duplex against thermal denaturation. The absorption melting profiles of the (C3T4C3) - (G3A4G3) x (G3A4G3) triplex are monophasic in agreement with previous reports. In contrast, the melting of this structure, when monitored by the pyrene excimer band, reveals a biphasic behavior. These data, together with kinetics measurements, strongly suggest exchange mechanisms between the homologous oligomers (G3A4G3), Hoogsteen, and Watson-Crick strands. PMID- 9398206 TI - Interaction of Alzheimer beta-amyloid peptide(1-40) with lipid membranes. AB - The beta-amyloid peptide beta AP(1-40), a 40-amino acid residues peptide, is one of the major components of Alzheimer's amyloid deposits. beta AP(1-40) exhibits only a limited solubility in aqueous solution and undergoes a concentration dependent, cooperative random coil reversible beta-structure transition for Cpep > 10 microM [Terzi, E., Holzemann, G., and Seelig, J. (1995) J. Mol. Biol. 252, 633-642]. In the presence of acidic lipid, the equilibrium is shifted further toward beta-structured aggregates. We have now characterized the lipid-peptide interaction using circular dichroism (CD) spectroscopy, lipid monolayers, and deuterium and phosphorus-31 solid-state nuclear magnetic resonance (NMR). CD spectroscopy revealed a distinct interaction between beta AP(1-40) and negatively charged unilamellar vesicles. In addition to the random coil reversible beta structured aggregate equilibrium at low lipid-to-peptide (L/P) ratios, a beta structure -->alpha-helix transition was observed at L/P > 55. beta AP(1-40) was found to insert into acidic monolayers provided the lateral pressure was low (20 mN/m). The extent of incorporation increased distinctly with the content of acidic lipid in the monolayer. However, at a lipid packing density equivalent to that of a bilayer (lateral pressure > or = 32 mN/m), no insertion of beta AP(1 40) was observed. The lipid molecular structure in the presence of beta AP(1-40) was studied with NMR. Phosphatidylcholine (PC) was selectively deuterated at the choline headgroup and at the cis-double bond of the oleic acyl chain and mixed with phosphatidylglycerol (PG). Phosphorus-31 NMR showed that the lipid phase retained the bilayer structure at all lipid-to-protein ratios. Deuterium NMR revealed no change in the headgroup conformation of the choline moiety or in the flexibility and ordering of the hydrocarbon chains upon the addition of beta AP (1-40). It can be concluded that beta AP(1-40) binds electrostatically to the outer envelope of the polar headgroup region without penetrating between the polar groups. The data suggest a new mechanism of helix formation induced by the proper alignment of five positive charges of beta AP(1-40) on the negatively charged membrane template. PMID- 9398207 TI - Inequivalence of the two tyrosine ligands in the N-lobe of human serum transferrin. AB - Human serum transferrin N-lobe (hTF/2N) has four iron-binding ligands, including one histidine, one aspartate, and two tyrosines. The present report elucidates the inequivalence of the two tyrosine ligands (Tyr 95 and Tyr 188) on the metal binding properties of hTF/2N by means of site-directed mutagenesis, metal release kinetics, and absorption and electron paramagnetic resonance (EPR) spectroscopies. When the liganding tyrosines were mutated individually to phenylalanine, the resulting mutant Y95F showed a weak binding affinity for iron and no affinity for copper, whereas, mutant Y188F completely lost the ability to bind iron but formed a stable complex with copper. Since other studies have demonstrated that mutations of the other two ligands, histidine and aspartate, did not completely abolish iron binding, the present findings suggest that the tyrosine ligand at position 188 is essential for binding of iron to occur. Replacement of Tyr 188 with phenylalanine created a favorable chemical environment for copper coordination but a fatal situation for iron binding. The positions of the two liganding tyrosines in the metal-binding cleft suggest a reason for the inequivalence. PMID- 9398209 TI - Cytosine methylation enhances DNA damage produced by groove binding and intercalating enediynes: studies with esperamicins A1 and C. AB - Methylation of the C5 position of cytosine in CG dinucleotides represents an important element in the control of gene expression in eukaryotic cells. This major groove modification of DNA causes changes in DNA conformation that are recognized by DNA-binding proteins and DNA-damaging chemicals. We have observed that CG methylation affects the DNA damage produced by the enediyne esperamicin A1 and its analog lacking the intercalating anthranilate, esperamicin C. Fragments of the human phosphoglycerate kinase gene (PGK1) and the plasmid pUC19 were methylated with SssI methylase and subjected to damage by esperamicins A1 and C. Damage produced by esperamicin A1 was enhanced 1.5-2-fold near a single CG sequence at position -101 in PGK1 and in a region containing several methylated CG dinucleotides between positions -120 and -131. Esperamicin C-induced damage was enhanced to a similar degree in PGK1 but only at the site that contained multiple CG dinucleotides. There was enhancement of damage for both drugs in the pUC19 fragment at several sites near CG sequences. Analysis of the chemistry of esperamicin-induced DNA damage suggests that cytosine methylation does not affect the identity of drug-abstracted hydrogen atoms. The damage chemistry was also used to identify the DNA binding orientation of the esperamicins. The anthranilate of esperamicin A1 is predicted to intercalate in a CT step four base pairs in a 3'-direction to the CG sequence at -101 in PGK1 and in a CG dinucleotide at the site containing multiple CGs (positions -120 to -131). These observations are consistent with other studies that indicate a long range effect of cytosine methylation on DNA conformation. PMID- 9398208 TI - Characterization of a covalent monoadduct of neocarzinostatin chromophore at a DNA bulge. AB - Neocarzinostatin chromophore (NCS-Chrom) induces highly efficient site-specific strand cleavage at the bulge of a folded single-stranded 31-mer DNA in the presence of oxygen [Kappen, L. S., and Goldberg, I. H. (1993) Science 261, 1319 1321]. Under anaerobic conditions, the major product is a material having gel mobility slower than that of the parent 31-mer. In order to characterize this product, it was stabilized by reduction with borane/pyridine, labeled with 32P at its 5' or 3' end, and subjected to chemical cleavage dependent on base elimination or modification, and the cleavage products were analyzed on a sequencing gel. A cleavage pattern comparable to that of the 31-mer was obtained until the bases on either side of T22 at the bulge. Cleaved fragments inclusive of T22 from the 5' or the 3' end had retarded and anomalous mobilities and appeared as a smear of bands closer to the starting material, presumably due to the presence of the covalently bound drug. Pyrimidine-specific agents such as hydrazine and potassium permanganate, but not the DNA sugar-specific probe thiol activated NCS-Chrom, induced strand cleavage at T22. Mass spectral analysis of the presumed adduct isolated from anaerobic reactions containing NCS-Chrom and a bulge duplex substrate made up of a 10-mer and an 8-mer showed that the adduct contains one molecule of the drug and one molecule of the 10-mer. Taken together, the results show that (i) drug adduction is at T22 on the full-length substrate; (ii) the pyrimidine ring is accessible to base-specific chemical modifications, hence, presumably free of the drug; (iii) it is most likely that drug adduction is via its C6 position to the 5' carbon of T22, based on the current results and the known chemistry of the hydrogen abstraction by the drug in the presence or absence of oxygen; (iv) there is no involvement of the neighboring bases by way of inter- or intrastrand cross-linking; and (v) the product is a monoadduct. PMID- 9398210 TI - Expression, Purification, and Inhibitory Properties of Human Proteinase Inhibitor 8 AB - In a previous report, the cDNA for human proteinase inhibitor 8 (PI8) was first identified, isolated, and subcloned into a mammalian expression vector and expressed in baby hamster kidney cells. Initial studies indicated that PI8 was able to inhibit the amidolytic activity of trypsin and form an SDS-stable approximately 67-kDa complex with human thrombin [Sprecher, C. A., et al. (1995) J. Biol Chem. 270, 29854-29861]. In the present study, we have expressed recombinant PI8 in the methylotropic yeast Pichia pastoris, purified the inhibitor to homogeneity, and investigated its ability to inhibit a variety of proteinases. PI8 inhibited the amidolytic activities of porcine trypsin, human thrombin, human coagulation factor Xa, and the Bacillus subtilis dibasic endoproteinase subtilisin A through different mechanisms but failed to inhibit the Staphylococcus aureus endoproteinase Glu-C. PI8 inhibited trypsin in a purely competitive manner, with an equilibrium inhibition constant (Ki) of less than 3.8 nM. The interaction between PI8 and thrombin occurred with a second-order association rate constant (kassoc) of 1.0 x 10(5) M-1 s-1 and a Ki of 350 pM. A slow-binding kinetics approach was used to determine the kinetic constants for the interactions of PI8 with factor Xa and subtilisin A. PI8 inhibited factor Xa via a two-step mechanism with a kassoc of 7.5 x 10(4) M-1 s-1 and an overall Ki of 272 pM. PI8 was a potent inhibitor of subtilisin A via a single-step mechanism with a kassoc of 1.16 x 10(6) M-1 s-1 and an overall Ki of 8.4 pM. The interaction between PI8 and subtilisin A may be of physiological significance, since subtilisin A is an evolutionary precursor to the intracellular mammalian dibasic processing endoproteinases. PMID- 9398211 TI - Human alpha 2-macroglobulin is an osteogenic growth peptide-binding protein. AB - The osteogenic growth peptide (OGP) is a 14mer mitogen of osteoblastic and fibroblastic cells. Physiologically, OGP is present in high abundance in human and other mammalian sera. Most of the serum OGP is complexed noncovalently to heat sensitive, high molecular weight OGP-binding proteins (OGPBPs). Changes in serum OGP levels that follow bone marrow ablation and the low doses of exogenous OGP required for the stimulation of bone formation suggest a regulatory role for the OGPBPs. In the present work, the OGP binding activity was monitored by competitive binding to [3-125I(Tyr10)]-sOGP and the corresponding complexes were demonstrated on nondenaturing cathodic polyacrylamide gel electrophoresis. We show that OGP binds to both native and activated human plasma alpha 2 macroglobulin (alpha 2M). alpha 2M was also immunoidentified in reduced and nonreduced SDS-polyacrylamide gel electrophoresis of OGP-affinity purified plasma derived proteins. Immunoreactive OGP was detected in commercial preparations of both forms of alpha 2M; OGP was purified to homogeneity from the commercial preparation of activated alpha 2M. In MC3T3 E1 cells, native alpha 2M, at concentrations < 50 ng/mL, had a substantially increased mitogenic effect in the presence of synthetic, native-like, OGP (sOGP). Similar amounts of activated alpha 2M inhibited the sOGP proliferative effect. These results suggest that the native alpha 2M enhances the immediate availability of OGP to its target cells. Activated alpha 2M may participate in the removal of OGP from the system. PMID- 9398212 TI - Persistence of dimerization and enzymatic activity of epidermal growth factor receptor in the absence of epidermal growth factor. AB - Dimerized and enzymatically active epidermal growth factor receptor, in the presence of saturating epidermal growth factor (EGF), was passed over a column containing an immunoadsorbent for EGF. The immunoadsorbent removed both EGF free in solution and EGF bound to EGF receptor, while EGF receptor passed through unimpeded. Both the dimerization of EGF receptor and the activation of its tyrosine kinase were arrested in samples containing EGF receptor that had been mixed with saturating EGF and then immediately passed over the immunoadsorbent for EGF. The EGF receptor in these samples, however, dimerized completely, and its tyrosine kinase became fully active upon readdition of EGF. Samples of EGF receptor that were fully dimerized and enzymatically active prior to being passed over the immunoadsorbent for EGF remained dimerized and enzymatically active even in the absence of bound EGF. The first-order rate constant for the inactivation of the tyrosine kinase of EGF receptor depleted of EGF was estimated by subtracting the rate constant of inactivation of the tyrosine kinase in samples replenished with EGF from the rate constant of inactivation of the tyrosine kinase in samples that had been depleted of EGF. The rate constant of inactivation was found to be 0.26 +/- 0.06 h-1. PMID- 9398213 TI - Altered ligand binding properties and enhanced stability of a constitutively active estrogen receptor: evidence that an open pocket conformation is required for ligand interaction. AB - To elucidate the ligand binding properties of the estrogen receptor (ER) and how ligand access to and release from the ligand binding pocket is affected by the conformational state of the receptor, we have measured the rates of estradiol association and dissociation, the equilibrium binding, and the stability of estradiol binding to denaturants, comparing wild-type human ER and a point mutant (Y537S ER) that shows full constitutive activity, i.e., the same full transcriptional activity in the absence or presence of estrogen. Ligand binding kinetics and affinity were measured with the full-length (1-595) ERs and with truncated forms of both receptors containing domains C through F (including the DNA binding, hinge, and ligand binding domains, amino acids 175-595) or domains E and F (the ligand binding domain; amino acids 304-595). With all ERs, the rates of ligand association and dissociation were considerably slower with the Y537S mutant ER than with wild-type ER (6-fold and 3-4-fold, respectively). These marked differences in ligand on and off rates for the wild-type and Y537S receptors result in a predicted (k-1/k+1) and measured Kd that is 2-fold lower for Y537S ER compared to wild-type ER. The binding of estradiol by wild-type ER was disrupted by high concentrations of urea (above 2 M), whereas the Y537S ER was distinctly more resistant to this disruption. These results are consistent with a model in which wild-type ER in the absence of ligand adopts a transcriptionally inactive collapsed pocket conformation, stabilized by specific interactions of Y537 with nearby regions of ER. When estradiol is bound, the wild type ER adopts a transcriptionally active, closed pocket (ligand occupied) conformation. By contrast, the Y537S mutant ER favors the transcriptionally active closed pocket conformation, whether occupied by ligand or not, the latter state (closed pocket but unoccupied) accounting for its constitutive activity. Our findings suggest that the entry or exit of ligand from the binding pocket requires that ER adopt an open pocket conformation. The reduced rates of ligand association and dissociation in the constitutively active form of the ER, as well as its greater resistance to disruption of ligand binding by urea, support the supposition that the rate at which this open pocket conformation can be accessed from the unoccupied or ligand-occupied Y537S ER is slower than from the unoccupied or occupied forms of wild-type ER. Thus, the binding and release of ligand by ER require that the receptor access an open pocket state, and the ease with which this state can be accessed is affected by mutations that alter receptor conformation. PMID- 9398214 TI - Structure and interaction of PA63 and EF (edema toxin) of Bacillus anthracis with lipid membrane. AB - The secondary structures of the two components of the Bacillus anthracis edema toxin, protective antigen (PA63) and edema factor (EF), as well as the two EF mutants: CYA30 (containing the N-terminal PA63-binding domain) and CYA62 (containing the C-terminal catalytic domain) were investigated as a function of pH in the absence and in the presence of phospholipid vesicles using attenuated total reflection Fourier transform infrared spectroscopy. Secondary structures were independent of pH, whereas, in all cases, structural modifications were observed upon lipid binding. The ability of PA63 and EF to undergo hydrogen/deuterium exchange was evaluated. The binding of these proteins and the mutants to the lipid membrane was also characterized and it was demonstrated that the association of PA63 to the lipid bilayer was pH-dependent, while the binding of EF to the lipid membrane took place at both neutral and acidic pH. Interestingly, the two EF mutants are showing different lipid binding properties in response to pH: CYA30 has a strong pH-dependence whereas CYA62, as EF, binds to the lipid vesicles at all pHs. For the two proteins characterized by a pH dependent lipid binding, the reversibility of binding upon neutralization was tested and binding of PA63 to the membrane was found to be irreversible whereas that of CYA30 was reversible. PMID- 9398215 TI - Determination of tumor necrosis factor binding protein disulfide structure: deviation of the fourth domain structure from the TNFR/NGFR family cysteine-rich region signature. AB - Tumor necrosis factor binding protein is a soluble molecule derived from the extracellular domain of the 55 kDa human tumor necrosis factor receptor, which can block the biological function of tumor necrosis factor by binding to the growth factor. This cysteine-rich molecule is subdivided into four domains, each containing six conserved cysteines that form three intrachain disulfide linkages known as the tumor necrosis factor receptor/nerve growth factor receptor family cysteine-rich region signature structure. In an effort to elucidate the molecular integrity of the molecule, we performed detailed analysis and searched for strategies to elucidate the complete disulfide structure of the E. coli-derived tumor necrosis factor binding protein and to determine the disulfide arrangement in the fourth domain of Chinese hamster ovary cell-derived molecule. The methods employed included various proteolytic digestions, peptide mapping, partial reduction, and assignment of disulfides by N-terminal sequencing and matrix assisted laser desorption ionization mass spectrometry with post-source decay. The first three domains of the molecule were confirmed to have disulfide structures identical to the cysteine-rich region signature structure found in the above-mentioned receptor superfamily. The fourth domain has a different structure from the first three domains where the last four cysteines form two disulfide bonds in opposite positions. PMID- 9398216 TI - The low-affinity ATP binding site of the Escherichia coli SecA dimer is localized at the subunit interface. AB - The homodimeric SecA protein is the ATP-dependent force generator in the Escherichia coli precursor protein translocation cascade. SecA contains two essential nucleotide binding sites (NBSs), i.e., NBS1 and NBS2 that bind ATP with high and low affinity, respectively. The photoactivatable bifunctional cross linking agent 3'-arylazido-8-azidoadenosine 5'-triphosphate (diN3ATP) was used to investigate the spatial arrangement of the nucleotide binding sites of SecA. DiN3ATP is an authentic ATP analogue as it supports SecA-dependent precursor protein translocation and translocation ATPase. UV-induced photo-cross-linking of the diN3ATP-bound SecA results in the formation of stable dimeric species of SecA. D209N SecA, a mutant unable to bind nucleotides at NBS1, was also photo cross-linked by diN3ATP, whereas no cross-linking occurred with the NBS2 mutant R509K SecA. We concluded that the low-affinity NBS2, which is located in the carboxyl-terminal half of SecA, is the site of crosslinking and that NBS2 binds nucleotides at or near the subunit interface of the SecA dimer. PMID- 9398217 TI - Evidence for a glutathionyl-enzyme intermediate in the amidase activity of the bifunctional glutathionylspermidine synthetase/amidase from Escherichia coli. AB - Glutathionylspermidine (Gsp) is a metabolite common to Escherichia coli and protozoal parasites of the Trypanosoma family. Though its role in E. coli is unknown, Gsp is known to be an intermediate in the biosynthesis of N1,N8 bis(glutathionyl)spermidine (trypanothione), a metabolite unique to trypanosomatids that may allow the parasites to overcome oxidative stresses induced by host defense mechanisms. The bifunctional Gsp-synthetase/amidase from E. coli catalyzes both amide bond formation and breakdown between the N1-amine of spermidine [N-(3-aminopropyl)-1,4-diaminobutane] and the glycine carboxylate of glutathione (gamma-Glu-Cys-Gly), with net hydrolysis of ATP [Bollinger et al. (1995) J. Biol. Chem. 270 (23), 14031-14041]. Synthetase and amidase activities reside in separate domains of the protein, and liberation of the amidase domain from the synthetase domain activates the amidase activity as much as 70-fold in kcat/K(m) for a chromogenic substrate gamma-Glu-Ala-Gly-pNA [Kwon et al., (1997) J. Biol. Chem. 272 (4), 2429-2436]. When substrates for the Gsp-synthetase activity are present (GSH, ATP-Mg2+), Gsp-amidase is highly activated (15-fold). We provide kinetic and mutagenesis evidence suggesting that the amidase operates by a nucleophilic attack mechanism involving cysteine as the catalytic nucleophile. Stopped-flow studies on the 25 kDa Gsp-amidase fragment and the 70 kDa full-length Gsp-synthetase/amidase with gamma-Glu-Ala-Gly-ONp demonstrate burst kinetics characteristic of a covalent acyl-enzyme intermediate. Studies using various group-specific protease inhibitors, such as iodoacetamide, suggest an active-site cysteine or histidine as being relevant to amidase activity, and site-directed mutagenesis indicates that Cys-59 is essential for amidase activity. PMID- 9398218 TI - Adenosylcobalamin-dependent glutamate mutase: examination of substrate and coenzyme binding in an engineered fusion protein possessing simplified subunit structure and kinetic properties. AB - Glutamate mutase is comprised of two weakly associating subunits; E and S, that combine to form the coenzyme binding site. The active holoenzyme assembles in a kinetically complex process in which both the stoichiometry and apparent Kd for adenosylcobalamin (AdoCbl) are dependent upon the relative concentrations of the two subunits, as is the enzyme's specific activity. To facilitate mechanistic and structural studies on this enzyme we have genetically fused the S subunit to the C-terminus of the E subunit through an 11 amino acid (Gly-Gln)5-Gly linker segment. This protein, GlmES, binds AdoCbl stoichiometrically and neither the affinity for AdoCbl nor the turnover number depends upon protein concentration. The kcat and Km for both substrate and coenzyme, together with the deuterium isotope effects on Vmax and Vmax/Km, have been determined for the GlmES-catalyzed reaction proceeding in both directions. Compared with wild type, the affinity for AdoCbl is unchanged, but for the conversion of L-glutamate to (2S,3S)-3 methylaspartate both kcat and Km for L-glutamate are decreased by about a third and the isotope effects are reduced, suggesting product release to be more rate limiting. To test hypotheses concerning the activation of the coenzyme, we examined the binding of adenosylcobalamin, methylcobalamin, and cob(II)alamin to the enzyme. Each of these is bound with essentially the same affinity (2 microM), suggesting that, contrary to expectations, interactions between the protein and the adenosyl moiety do not serve to weaken the cobalt-carbon bond in the ground state. PMID- 9398219 TI - Minimizing nonproductive substrate binding: a new look at glucoamylase subsite affinities. AB - A subsite model as proposed by Hiromi [Hiromi, K. (1970) Biochem. Biophys. Res. Commun. 40, 1-6] has been applied to various hydrolases including glucoamylase (GA). The model assumes a single enzyme complex, a hydrolytic rate constant which is independent of substrate length, and a ratelimiting hydrolytic step. Recent kinetic studies with GA contradict these assumptions. Here we reevaluate the substrate binding of GA studying the pre-steady-state kinetics with glucose, which is reported here for the first time, and maltose. The association equilibrium constants for glucose and maltose interactions with wild-type and Trp120-->Phe GA from Aspergillus awamori in H2O and D2O buffers were obtained. Kinetic results indicate that a single glucose molecule binds to GA weakly by a single-step mechanism, E + G1<-->EG1, under the conditions studied. Similar fluorescence intensities of the GA-glucose and GA-maltose complexes, the high tryptophan concentration around subsite 1, crystal structures of various inhibitor complexes, pre-steady-state and steady-state modeling, feasibility of condensation reactions, and other evidence strongly suggest that glucose binds at subsite 1. These results conflict with the high subsite 2 and low subsite 1 affinities obtained using Hiromi's model. Using the substrate association constants for glucose and maltose obtained by pre-steady-state kinetics, the affinity of alpha-glucose for subsite 1 is shown to be substantially higher than the apparent affinity of glucose for subsite 2. We propose a GA catalytic mechanism whereby substrate binding is initiated by subsite 1 interactions with the nonreducing end of the oligosaccharide substrate, minimizing nonproductive substrate binding. Through conformational changes, entropic contributions, and increased local concentration, subsite 2 subsequently has enhanced affinity for the second covalently linked glucosyl residue. PMID- 9398220 TI - Role of mitochondrial and cytoplasmic serine hydroxymethyltransferase isozymes in de novo purine synthesis in Saccharomyces cerevisiae. AB - One-carbon units are essential to a variety of anabolic processes which yield necessary cellular components including purines, pyrimidines, amino acids, and lipids. Serine hydroxymethyltransferase (SHMT) is the major provider of one carbon units in the cell. The other product of this reaction is glycine. Both of these metabolites are required in de novo purine biosynthesis. In Saccharomyces cerevisiae, mitochondrial and cytoplasmic SHMT isozymes are encoded by distinct nuclear genes (SHM1 and SHM2). Molecular genetic analyses have begun to define the roles of these two isozymes in folate-mediated one-carbon metabolism [McNeil, J. B., et al. (1996) Genetics 142, 371-381]. In our study, the SHM1 and SHM2 genes were disrupted singly and in combination to investigate the contributions of the two SHMT isozymes to the production of glycine and one-carbon units required in purine biosynthesis. Cell subfractionation experiments indicated that while only 5% of total activity was localized in the mitochondria, the specific activity in that compartment was much higher than in the cytoplasm. Growth and 13C NMR experiments indicate that the two isozymes function in different directions, depending on the nutritional conditions of the cell. When yeast was grown on serine as the primary one-carbon source, the cytoplasmic isozyme was the main provider of glycine and one-carbon groups for purine synthesis. When grown on glycine, the mitochondrial SHMT was the predominant isozyme catalyzing the synthesis of serine from glycine and one-carbon units. However, when both serine and glycine were present, the mitochondrial SHMT made a significant contribution of one-carbon units, but not glycine, for purine synthesis. Finally, NMR data are presented that suggest the existence of at least two sites of de novo purine biosynthesis in growing yeast cells, each being fed by distinct pools of precursors. PMID- 9398221 TI - Catalytic mechanism of phosphorylation and dephosphorylation of CheY: kinetic characterization of imidazole phosphates as phosphodonors and the role of acid catalysis. AB - Kinetic and equilibrium measurements of phosphotransfer events involving CheY carried out over a range of pH conditions elucidated several features of the phosphotransfer mechanism. Using tryptophan fluorescence intensity measurements as a monitor of phosphorylation, we showed that phosphorylation using small molecule phosphodonors occurred by fast association of CheY with the phosphodonor, followed by rate-limiting phosphotransfer. Two previously uncharacterized phosphodonors, monophosphoimidazole and diphosphoimdazole, were able to phosphorylate CheY at a concentration about 6-fold lower than that of the previously described phosphodonors acetyl phosphate and phosphoramidate. This was shown to be due to tighter binding of the imidazole phosphates to CheY and implied the presence of binding interactions between CheY and the imidazole group. The ability of CheY to autophosphorylate through the pH range of 5-10 differed for various phosphodonors. Acetyl phosphate and diphosphoimidazole were unaffected by pH over this range, whereas phosphoramidate and monophosphoimidazole showed a steep dependence on pH with a loss of phosphorylation ability at about pH 7.4 (midpoint) for monophosphoimidazole and pH 7.8 (midpoint) for phosphoramidate. This behavior correlated with the loss of the positive charge on the nitrogen atom in the nitrogen-phosphorus bond in both monophosphoimidazole and phosphoramidate and implied that CheY was not capable of donating a proton to the leaving group in phosphotransfer with small molecules. The rate of phosphotransfer from [32P]CheA-phosphate to wild type CheY also decreased markedly (> 150 times) between pH 7.5 and 10. Because the mutant CheY proteins K109R and T87A showed the same pH dependence as the wild type, the loss of activity in the alkaline range could not be attributed to deprotonation of either of these active site residues. This observation, combined with the moderate decreases in phosphotransfer rates for these mutants relative to that of wild type CheY, indicated that it is unlikely that either Thr87 or Lys109 plays a direct role in the catalysis of phosphotransfer. Finally, we showed that the rate of autodephosphorylation of CheY was independent of pH over the range of 4.5-11. Together, these studies led to a model with CheY playing a largely entropic role in its own phosphorylation and dephosphorylation. PMID- 9398222 TI - Mechanism of Formation of Novel Covalent Drug·DNA Interstrand Cross-Links and Monoadducts by Enediyne Antitumor Antibiotics AB - The potent enediyne antitumor antibiotic C1027 has been previously reported to induce novel DNA interstrand cross-links and drug monoadducts under anaerobic conditions [Xu et al. (1997) J. Am. Chem. Soc. 119, 1133-1134]. In the present study, we explored the mechanism of formation of these anaerobic DNA lesions. We found that, similar to the aerobic reaction, the diradical species of the activated drug initiates anaerobic DNA damage by abstracting hydrogen atoms from the C4', C1', and C5' positions of the A1, A2, and A3 nucleotides, respectively, in the most preferred 5'GTTA1T/5'ATA2A3C binding sequence. It is proposed that the newly generated deoxyribosyl radicals, which cannot undergo oxidation, likely add back onto the nearby unsaturated ring system of the postactivated enediyne core, inducing the formation of interstrand cross-links, connecting either A1 to A2 or A1 to A3, or drug monoadducts mainly on A2 or A3. Comparative studies with other enediynes, such as neocarzinostatin and calicheamicin gamma1I under similar reaction conditions indicate that the anaerobic reaction process is a kinetically competitive one, depending on the proximity of the drug unsaturated ring system or dioxygen to the sugar radicals and their quenching by other hydrogen sources such as solvent or thiols. It was found that C1027 mainly generates interstrand cross-links, whereas most of the anaerobic lesions produced by neocarzinostatin are drug monoadducts. Calicheamicin gamma1I was found to be less efficient in producing both lesions. The anaerobic DNA lesions induced by enediyne antitumor antibiotics may have important implications for their potent cytotoxicity in the central regions of large tumors, where relative anaerobic conditions prevail. PMID- 9398223 TI - Microscopic pKa values of Escherichia coli thioredoxin. AB - Thiol:disulfide oxidoreductases have a CXXC motif within their active sites. To initiate the reduction of a substrate disulfide bond, the thiolate form of the N terminal cysteine residue (CXXC) of this motif performs a nucleophilic attack. Escherichia coli thioredoxin [Trx (CGPC)] is the best characterized thiol:disulfide oxidoreductase. Previous determinations of the active-site pKa values of Trx have led to conflicting interpretations. Here, 13C-NMR spectroscopy, site-specific isotopic labeling, and site-directed mutagenesis were used to demonstrate that analysis of the titration behavior of wild-type Trx requires the invocation of microscopic pKa values for two interacting active-site residues: Asp26 (7.5 and 9.2) and Cys32 (CXXC; 7.5 and 9.2). By contrast, in two Trx variants, D26N Trx and D26L Trx, Cys32 exhibits a pKa near 7.5 and has a well defined, single-pKa titration curve. Similarly, in oxidized wild-type Trx, Asp26 has a pKa near 7.5. In CVWC and CWGC Trx, Cys32 exhibits a single pKa near 6.2. In all five enzymes studied here, there is no evidence for a Cys35 (CXXC) pKa of < 11. This study demonstrates that a comprehensive approach must be used to unravel complex titration behavior of the functional groups in a protein. PMID- 9398224 TI - Kinetics of cytochrome c folding examined by hydrogen exchange and mass spectrometry. AB - Pulsed hydrogen exchange/mass spectrometry, a new method for studying protein folding, has been used to investigate folding of cytochrome c on the 5 ms to 15 s time scale. Cytochrome c, unfolded in guanidine hydrochloride/D2O, was allowed to refold in a high-speed quenched-flow apparatus and pulse-labeled with protium to identify unfolded regions. Intact, labeled cytochrome c was digested into fragments which were analyzed by HPLC electrospray ionization mass spectrometry to determine the level of deuterium in each fragment. Bimodal distributions of deuterium were found for most segments, indicating that regions represented by these segments were either unfolded or completely folded in the intact polypeptide prior to labeling. This behavior is consistent with cooperative, localized folding which occurs in less than 10 ms in individual molecules. Deuterium levels found in the fragments were normalized to levels found in the same fragments derived from folded cytochrome c, pulse-labeled in the same manner, to indicate the percentage of cytochrome c that was folded. These results show that the N/C-terminal regions fold cooperatively on a time scale extending from less than the mixing time of the apparatus (5 ms) to as long as 15 s, and that the other regions also fold cooperatively. However, these regions do not begin to fold until 30 ms after mixing. In addition to providing new information on cytochrome c folding, these results demonstrate that pulse-hydrogen exchange/mass spectrometry is complementary to NMR in some respects and advantageous in others. Results of this study form the foundation required to extend the pulsed hydrogen exchange approach to folding studies of proteins too large to be analyzed by NMR. PMID- 9398225 TI - Intersubunit communication in hybrid hexamers of K89L/A163G/S380A and C320S mutants of glutamate dehydrogenase from Clostridium symbiosum. AB - The triple mutant K89L/A163G/S380A (inactive with glutamate but active with L-Nle and L-Met) and C320S (fully active with glutamate, entirely inactive with L-Nle and L-Met, and also lacking reactive cysteine) mutant of glutamate dehydrogenase (EC 1.4.1.2) of Clostridium symbiosum could be completely denatured by urea with the loss of structure and activity. The mutants denatured by urea could be reassociated to give stable hexamers with recovery of activity of approximately 67% by dilution in 0.1 M potassium phosphate buffer (pH 7.0) containing 2 mM NAD+. The native, urea-denatured, and renatured states of mutant enzymes were characterized by size exclusion chromatography on FPLC and native PAGE. Intersubunit hybrid hexamers containing five subunits of triple mutant and one subunit of C320S mutant were constructed by in vitro subunit hybridization followed by affinity chromatography. Kinetic analysis showed that a 5:1 hybrid hexamer, with only one C320S subunit able to bind NAD+ after DTNB modification, shows classical Michaelis-Menten kinetics with regard to NAD+. This contrasts with the apparent negative co-operativity shown by pure C320S hexamers and suggests that the interaction in NAD+ binding among subunits is eliminated in the hybrid. After removal of thionitrobenzoate, however, all of the subunits in the hybrid are able to bind NAD+. In this state the hybrid enzyme showed slight deviation from classical behavior with regard to NAD+, indicating reintroduction of some level of allosteric interaction. The hybrid hexamer also showed much reduced co-operativity with glutamate at pH 8.8, with a Hill coefficient of 3 for DTNB-treated hybrid (as compared to 5.2 for the pure C320S mutant) and 2.2 for the untreated hybrid. The fact that co-operativity in glutamate binding is not entirely eliminated correlates with evidence that the triple mutant subunits, though inactive toward glutamate, can nevertheless still bind this amino acid. PMID- 9398226 TI - Fast events in protein folding: relaxation dynamics and structure of the I form of apomyoglobin. AB - The fast relaxation dynamics of the acid destabilized I form of apomyoglobin (pH* 3, 0.15 M NaCl; apoMb-I) following a laser-induced temperature-jump have been probed using time-resolved infrared spectroscopy. Only a fast, single exponential phase is observed (bleach centered at v = 1633 cm-1 and transient absorbance at 1666 cm-1) with relaxation times of 38 ns at 30 degrees C and 36 ns at 57 degrees C; no additional slow (microsecond) phase is observed as previously found in the native form of apomyoglobin. Folding times of approximately 66 ns are derived from the observed rates based on a simple two-state model. The equilibrium melting of the 1633 cm-1 component shows noncooperative linear behavior over the temperature range studied (10-60 degrees C). The low amide I' frequency, the fast relaxation dynamics, and the noncooperative melting behavior are characteristic of isolated solvated helix. The analysis of the amide-I' band reveals another major component at 1650 cm-1 assigned to native-like structure stabilized by tertiary contacts involving the AGH core, which does not show dynamic or static melting under our conditions. ApoMb-I has generally been taken to be a "molten globule" species. The present results indicate a heterogeneous structure consisting of separate regions of native-like unit(s), solvated helices, and disordered coil, excluding a homogeneous molten globule as a model for apoMb-I. From the current studies and other results, a detailed model of the folding of apomyoglobin is presented. PMID- 9398227 TI - Novel application of 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole to identify cysteine sulfenic acid in the AhpC component of alkyl hydroperoxide reductase. AB - The trapping of a sulfenic acid within the fully active C165S mutant of the AhpC peroxidase protein from Salmonella typhimurium was investigated. The electrophilic reagent employed in these studies, 7-chloro-4-nitrobenz-2-oxa-1,3 diazole (NBD-Cl), has previously been used to modify thiol, amino, and tyrosine hydroxyl groups in proteins; at neutral pH only cysteinyl residues of AhpC proteins are modified. The peroxide-oxidized C165S mutant of AhpC incubated with NBD-Cl gave a product with an absorbance maximum at 347 nm, whereas the thiol-NBD conjugate formed from the reduced protein absorbed maximally at 420 nm. Electrospray ionization mass spectrometry of the modified proteins allowed identification of the species absorbing at 347 nm as a Cys-S(O)-NBD derivative containing one additional oxygen relative to the Cys-S-NBD product. The C165S conjugates with Cys-S(O)-NBD and Cys-S-NBD had no peroxidase activity when compared to unreacted C165S and wild-type AhpC, but were both reactivated through removal of NBD by DTT. Oxidized C165S was also modified by dimedone, a common sulfenic acid reagent, to give the expected inactivated conjugate of higher mass. This reagent was not removed by DTT and blocked any further reaction of the protein with NBD-Cl. NBD modification of Enterococcus faecalis NADH peroxidase, a well-characterized flavoprotein with an active-site sulfenic acid (Cys-SOH), also yielded the spectrally-distinguishable NBD conjugates following incubation of NBD Cl with oxidized and reduced forms of the denatured peroxidase, indicating a general utility for this reagent with other sulfenic acid-containing proteins. A significant advantage of NBD-Cl over previously-used sulfenic acid reagents such as dimedone is in the retention of the sulfenic acid oxygen in the modified product; differentiation between protein-associated thiols and sulfenic acids is therefore now possible by means of both visible absorbance properties and mass analyses of the NBD-modified proteins. PMID- 9398228 TI - Identification of residues essential for differential fatty acyl specificity of Geotrichum candidum lipases I and II. AB - The fungus Geotrichum candidum produces two lipase isoenzymes, GCL I and GCL II, with distinct differences in substrate specificity despite their 86% identical primary structure. GCL I prefers ester substrates with long-chain cis (delta-9) unsaturated fatty acid moieties, whereas GCL II also accepts medium-length (C8 C14) acyl moieties in the substrate. To reveal structural elements responsible for differences in substrate differentiating ability of these isoenzymes, we designed, expressed, and characterized 12 recombinant lipase variants. Three chimeric lipases containing unique portions of the N-terminal and the C-terminal part of GCL I and GCL II, respectively, were constructed and enzymatically characterized. Activities were measured against mixed triglyceride-poly(dimethyl siloxane) particles. Our results indicate that residues within sequence positions 349-406 are essential for GCL I's high triolein/trioctanoin activity ratio of 20. The substitution of that segment in the specific GCL I to the corresponding residues in the nonspecific GCL II resulted in an enzyme with a triolein/trioctanoin activity ratio of 1.4, identical to that of GCL II. The reverse mutation in GCL II increased its specificity for triolein by a factor of 2, thus only in part restoring the high specificity seen with GCL I. In further experiments, the point mutations at the active site entrance of the GCL I, Leu358Phe and Ile357Ala/Leu358Phe, lowered the triolein/trioctanoin activity ratio from 20 to 4 and 2.5, respectively. The substitutions Cys379Phe/Ser380Tyr at the bottom of the active site cavity of GCL I decreased its specificity to a value of 3.6. Measurements of lipase activity with substrate particles composed of pure triglycerides or ethyl esters of oleic and octanoic acids resulted in qualitatively similar results as reported above. Our data reveal for the first time the identity of residues essential for the unusual substrate preference of GCL I and show that the anatomy, both at the entrance and the bottom of the active site cavity, plays a key role in substrate discrimination. PMID- 9398229 TI - Calmodulin binds to caldesmon in an antiparallel manner. AB - Two of the five tryptophan residues (W659 and W692) in chicken gizzard smooth muscle caldesmon (CaD) are located within the calmodulin (CaM) binding sites in the C-terminal region of the molecule. When these Trp residues are replaced with Gly in either recombinant fragments or synthetic peptides of CaD, the affinity for CaM is decreased by at least 10-fold, suggesting that both of these residues are important for the interaction of CaD with CaM. To gain information about the topography of the CaM-CaD complex, we have carried out fluorescence titrations of CaM with Tb3+ as a substitute for Ca2+ in the presence of wild-type or mutated CaD variants. By exciting Trp residues of CaD fragments or peptides while monitoring the enhanced luminescence of CaM-bound Tb3+ ions via resonance energy transfer, we were able to estimate the relative proximity between the bound metal ions in the two domains of CaM and the Trp residues of CaD. Our results suggest that in the CaM-CaD complex the metal-binding sites III and IV in the C-terminal domain of CaM are very close to W659 of CaD; the N-terminal domain of CaM appears associated with the region of CaD in the vicinity of W692, although sites I and II are relatively far away from this Trp residue. These findings are consistent with a model in which CaM binds to CaD in an antiparallel manner. Such a binding mode, however, may be flexible enough to accommodate alternative spatial arrangements when the preferred binding sites are either altered or rendered unavailable. PMID- 9398230 TI - Reconstruction of quaternary structures of class II tRNA synthetases by rational mutagenensis of a conserved domain. AB - Class II tRNA synthetases have long been known to have quaternary structures of alpha, alpha2, alpha2beta2, and alpha4, depending on the amino acid specificity and the organism from which the synthetase was isolated. Even the quaternary structures of enzymes for the same amino acid show variations in evolution. The basis for these variations has not been understood. We report here that sequence manipulations of a structural motif (motif 1) characteristic of all class II tRNA synthetases can generate most of the evolutionary diversity of quaternary forms of class II synthetases. Thus, the principles elucidated here for quaternary structure assembly may be general. PMID- 9398231 TI - Remarkable ability of horse spleen apoferritin to demetallate hemin and to metallate protoporphyrin IX as a function of pH. AB - In previous studies it has been shown that reaction of crystalline horse spleen apoferritin with hemin leads to a protoporphyrin IX-apoferritin complex [Precigoux et al. (1994) Acta Crystallogr. D50, 739-743]. We show here the following. (i) Hemin binds to two classes of sites in horse spleen apoferritin at pH 8, each with a binding stoichiometry of 0.5 hemin/subunit; protoporphyrin IX also binds to horse spleen apoferritin with an apparent binding stoichiometry of 1 molecule of protoporphyrin IX/subunit. (ii) When Fe(III)-protoporphyrin IX binds to apoferritin, there is a pH-dependent loss of the metal ion, extremely slow at alkaline pH values (half-time of weeks) and much more rapid at acidic pH values (half-time of seconds below pH 5.0); maximum rates of demetallation are found at pH 4.0, and at lower pH values they decrease. (iii) Chemical modification of 11 carboxyl groups/subunit in horse spleen apoferritin does not affect hemin binding at alkaline pH values; however, it prevents hemin demetallation at acidic pH values. (iv) Hemin that has been demetallated at acidic pH values can be remetallated by increasing the pH; the rate of remetallation is greater at more alkaline pH values. (v) When around 20 atoms of iron/molecule are incorporated into horse spleen apoferritin and protoporphyrin IX is then bound, iron can subsequently be transferred to the porphyrin at pH 8.0. A mechanism is proposed to explain demetallation of heme, involving attack on the tetrapyrrole nitrogens of the protoporphyrin IX-Fe by protons derived from protein carboxylic acid groups and subsequent complexation of the iron by the corresponding carboxylates and binding of protoporphyrin IX to a preformed pocket in the inner surface of the apoferritin protein shell. The cluster of carboxylates involved is situated at the entrance to the pocket in which the protoporphyrin IX molecule is bound and has been previously identified as the site of iron incorporation into L-chain apoferritins. This appears to be the first example of iron removal and incorporation into porphyrins under relatively mild physiological conditions. PMID- 9398232 TI - Site-directed spin-labeling of transmembrane domain VII and the 4B1 antibody epitope in the lactose permease of Escherichia coli. AB - Functional lactose permease mutants containing single Cys residues at positions 233-255 and a biotin acceptor domain at the C terminus were solubilized in dodecyl beta-d-maltopyranoside and purified by avidin affinity chromatography. Each mutant protein was derivatized with a thiol-selective nitroxide reagent and examined by conventional and power saturation electron paramagnetic resonance spectroscopy (EPR). The EPR spectral line shapes and the influence of nonpolar O2 or polar potassium chromium oxalate relaxation agents on the saturation behavior of the spin-labeled proteins were measured in order to obtain information on the mobility of the spin-labeled side chains and their accessibility to the relaxation agents, respectively. The results provide evidence that residues Ser233-Asn246 are within the hydrophobic core of the membrane and that Phe247 is at the lipid headgroup-solvent interface. Along with Phe247, Phe250 and Gly254 are also surface-exposed, as indicated by studies on the epitope for monoclonal antibody 4B1 [Sun, J., Wu, J., Carasco, N., and Kaback, H. R. (1996) Biochemistry 35, 990-998]. Furthermore, the nitroxide-labeled intramembrane Cys replacements exhibit variations in mobility and accessibility that are consistent with the conclusion that TM VII is an alpha-helix in contact with surrounding helices in the tertiary structure of the permease. PMID- 9398233 TI - Three-dimensional structure of leucocin A in trifluoroethanol and dodecylphosphocholine micelles: spatial location of residues critical for biological activity in type IIa bacteriocins from lactic acid bacteria. AB - The first three-dimensional structure of a type IIa bacteriocin from lactic acid bacteria is reported. Complete 1H resonance assignments of leucocin A, a 37 amino acid antimicrobial peptide isolated from the lactic acid bacterium Leuconostoc gelidum UAL187, were determined in 90% trifluoroethanol (TFE)-water and in aqueous dodecylphosphocholine (DPC) micelles (1:40 ratio of leucocin A:DPC) using two-dimensional NMR techniques (e.g., DQF-COSY, TOCSY, NOESY). Circular dichroism spectra, NMR chemical shift indices, amide hydrogen exchange rates, and long range nuclear Overhauser effects indicate that leucocin A adopts a reasonably well defined structure in both TFE and DPC micelle environments but exists as a random coil in water or aqueous DMSO. Distance geometry and simulated annealing calculations were employed to generate structures for leucocin A in both lipophilic media. While some differences were noted between the structures calculated for the two different solvent systems, in both, the region encompassing residues 17-31 assumes an essentially identical amphiphilic alpha helix conformation. A three-strand antiparallel beta-sheet domain (residues 2 16), anchored by the disulfide bridge, is also observed in both media. In TFE, these two regions have a more defined relationship relative to each other, while, in DPC micelles, the C-terminus is folded back onto the alpha-helix. The implications of these structural features with regard to the antimicrobial mechanism of action and target recognition are discussed. PMID- 9398234 TI - X-ray crystallographic studies of unique cross-linked lattices between four isomeric biantennary oligosaccharides and soybean agglutinin. AB - Soybean agglutinin (SBA) (Glycine max) is a tetrameric GalNAc/Gal-specific lectin which forms unique cross-linked complexes with a series of naturally occurring and synthetic multiantennary carbohydrates with terminal GalNAc or Gal residues [Gupta et al. (1994) Biochemistry 33, 7495-7504]. We recently reported the X-ray crystal structure of SBA cross-linked with a biantennary analog of the blood group I carbohydrate antigen [Dessen et al. (1995) Biochemistry 34, 4933-4942]. In order to determine the molecular basis of different carbohydrate-lectin cross linked lattices, a comparison has been made of the X-ray crystallographic structures of SBA cross-linked with four isomeric analogs of the biantennary blood group I carbohydrate antigen. The four pentasaccharides possess the common structure of (beta-LacNAc)2Gal-beta-R, where R is -O(CH2)5COOCH3. The beta-LacNAc moieties in the four carbohydrates are linked to the 2,3-, 2,4-, 3,6-, and 2,6 positions of the core Gal residue(s), respectively. The structures of all four complexes have been refined to approximately 2.4-2.8 A. Noncovalent lattice formation in all four complexes is promoted uniquely by the bridging action of the two arms of each bivalent carbohydrate. Association between SBA tetramers involves binding of the terminal Gal residues of the pentasaccharides at identical sites in each monomer, with the sugar(s) cross-linking to a symmetry related neighbor molecule. While the 2,4-, 3,6-, and 2,6-pentasaccharide complexes possess a common P6422 space group, their unit cell dimensions differ. The 2, 3-pentasaccharide cross-linked complex, on the other hand, possesses the space group I4122. Thus, all four complexes are crystallographically distinct. The four cross-linking carbohydrates are in similar conformations, possessing a pseudo-2-fold axis of symmetry which lies on a crystallographic 2-fold axis of symmetry in each lattice. In the case of the 3,6- and 2,6-pentasaccharides, the symmetry of their cross-linked lattices requires different rotamer orientations about their beta(1,6) glycosidic bonds. The results demonstrate that crystal packing interactions are the molecular basis for the formation of distinct cross linked lattices between SBA and four isomeric pentasaccharides. The present findings are discussed in terms of lectins forming unique cross-linked complexes with glycoconjugate receptors in biological systems. PMID- 9398235 TI - Three-dimensional structure of Escherichia coli dihydrodipicolinate reductase in complex with NADH and the inhibitor 2,6-pyridinedicarboxylate. AB - Dihydrodipicolinate reductase catalyzes the NAD(P)H-dependent reduction of the alpha,beta-unsaturated cyclic imine dihydrodipicolinate to form the cyclic imine tetrahydrodipicolinate. The enzyme is a component of the biosynthetic pathway that leads to diaminopimelate and lysine in bacteria and higher plants. Because these pathways are unique to microorganisms and plants, they may represent attractive targets for new antimicrobial or herbicidal compounds. The three dimensional structure of the ternary complex of Escherichia coli dihydrodipicolinate reductase with NADH and the inhibitor 2,6 pyridinedicarboxylate has been solved using a combination of molecular replacement and noncrystallographic symmetry averaging procedures and refined against 2.6 A resolution data to a crystallographic R-factor of 21.4% (Rfree is 29.7%). The native enzyme is a 120 000 molecular weight tetramer of identical subunits. The refined crystallographic model contains a tetramer, three molecules of NADH, three molecules of inhibitor, one phosphate ion, and 186 water molecules per asymmetric unit. Each subunit consists of two domains connected by two flexible hinge regions. While three of the four subunits of the tetramer have a closed conformation, in which the nicotinamide ring of the cofactor bound to the N-terminal domain and the reducible carbon of the substrate bound to the substrate binding domain are about 3.5 A away, the fourth subunit is unliganded and shows an open conformation, suggesting that the enzyme undergoes a major conformational change upon binding of both substrates. The residues involved in binding of the inhibitor and the residues involved in catalysis have been identified on the basis of the three-dimensional structure. Site-directed mutants have been used to further characterize the role of these residues in binding and catalysis. A chemical mechanism for the enzyme, based on these and previously reported data, is proposed. PMID- 9398236 TI - Structure of the carboxy-terminal fragment of the apo-biotin carboxyl carrier subunit of Escherichia coli acetyl-CoA carboxylase. AB - The biotin carboxyl carrier protein (BCCP) is a subunit of acetyl-CoA carboxylase, a biotin-dependent enzyme that catalyzes the first committed step of fatty acid biosynthesis. In its functional cycle the biotin carboxyl carrier protein engages in heterologous protein-protein interactions with three distinct partners, depending on its state of posttranslational modification. Apo-BCCP interacts specifically with the biotin holoenzyme synthetase, BirA, which results in the posttranslational attachment of biotin to an essential lysine residue on BCCP. Holo-BCCP then interacts with the biotin carboxylase subunit, which leads to the addition of the carboxylate group of bicarbonate to biotin. Finally, the carboxybiotinylated form of BCCP interacts with transcarboxylase in the conversion of acetyl-CoA to malonyl-CoA. The determinants of protein-protein interaction specificity in this system are unknown. One hypothesis is that posttranslational modification of BCCP may result in conformational changes that regulate specific protein-protein interactions. To test this hypothesis, we have determined the NMR solution structure of the unbiotinylated form of an 87 residue C-terminal domain fragment of BCCP (apoBCCP87) from Escherichia coli acetyl-CoA carboxylase and compared this structure with the high-resolution structure of the biotinylated form that was recently solved by X-ray crystallographic techniques. Although the overall folding of the two proteins is highly similar, small structural differences are apparent for residues of the biotin-binding loop that may be important for mediating specific protein-protein interactions. PMID- 9398237 TI - Identification and structural and functional characterization of human enamelysin (MMP-20). AB - A cDNA encoding a new human matrix metalloproteinase (MMP) has been cloned from RNA prepared from odontoblastic cells. The open reading frame of the cloned cDNA codes for a polypeptide of 483 amino acids and is extensively similar to the sequence of recently described porcine enamelysin, suggesting that the isolated cDNA codes for the human homologue of this enzyme. Human enamelysin (MMP-20) has a domain organization similar to other MMPs, including a signal peptide, a prodomain with the conserved motif PRCGVPD involved in maintaining enzyme latency, a catalytic domain with a Zn-binding site, and a COOH-terminal fragment similar to the sequence of hemopexin. The calculated molecular mass of human enamelysin is about 54 kDa, which is similar to that of collagenases or stromelysins. However, this human MMP lacks a series of structural features distinctive of these subfamilies of MMPs. The full-length human enamelysin cDNA has been expressed in Escherichia coli, and the purified and refolded recombinant protein is able to degrade synthetic peptides used as substrates of MMPs, confirming that human enamelysin belongs to this family of proteases. Furthermore, the recombinant human enamelysin is able to degrade amelogenin, the major protein component of the enamel matrix. On the basis of its degrading activity on amelogenin, and its highly restricted expression to dental tissues, we suggest that human enamelysin plays a central role in the process of tooth enamel formation. Finally, we have found that the human enamelysin gene (MMP-20) maps to chromosome 11q22, clustered to at least seven other members of the MMP gene family. PMID- 9398238 TI - Iron-sulfur cluster cysteine-to-serine mutants of Anabaena -2Fe-2S- ferredoxin exhibit unexpected redox properties and are competent in electron transfer to ferredoxin:NADP+ reductase. AB - The reduction potentials and the rate constants for electron transfer (et) to ferredoxin:NADP+ reductase (FNR) are reported for site-directed mutants of the [2Fe-2S] vegetative cell ferredoxin (Fd) from Anabaena PCC 7120, each of which has a cluster ligating cysteine residue mutated to serine (C41S, C46S, and C49S). The X-ray crystal structure of the C49S mutant has also been determined. The UV visible optical and CD spectra of the mutants differ from each other and from wild-type (wt) Fd. This is a consequence of oxygen replacing one of the ligating cysteine sulfur atoms, thus altering the ligand --> Fe charge transfer transition energies and the chiro-optical properties of the chromophore. Each mutant is able to rapidly accept an electron from deazariboflavin semiquinone (dRfH.) and to transfer an electron from its reduced form to oxidized FNR although all are somewhat less reactive (30-50%) toward FNR and are appreciably less stable in solution than is wt Fd. Whereas the reduction potential of C46S (-381 mV) is not significantly altered from that of wt Fd (-384 mV), the potential of the C49S mutant (-329 mV) is shifted positively by 55 mV, demonstrating that the cluster potential is sensitive to mutations made at the ferric iron in reduced [2Fe-2S] Fds with localized valences. Despite the decrease in thermodynamic driving force for et from C49S to FNR, the et rate constant is similar to that measured for C46S. Thus, the et reactivity of the mutants does not correlate with altered reduction potentials. The et rate constants of the mutants also do not correlate with the apparent binding constants of the intermediate (Fdred:FNRox) complexes or with the ability of the prosthetic group to be reduced by dRfH.. Furthermore, the X-ray crystal structure of the C49S mutant is virtually identical to that of wt Fd. We conclude from these data that cysteine sulfur d-orbitals are not essential for et into or out of the iron atoms of the cluster and that the decreased et reactivity of these Fd mutants toward FNR may be due to small changes in the mutual orientation of the proteins within the intermediate complex and/or alterations in the electronic structure of the [2Fe-2S] cluster. PMID- 9398239 TI - Substitution of the urease active site carbamate by dithiocarbamate and vanadate. AB - Urease possesses a dinuclear nickel active site with the metals bridged by a carbamylated lysine residue. In vitro activation of apoprotein (Apo) is achieved by incubation with Ni(II) and bicarbonate as a source of CO2. Analogues of CO2 and bicarbonate were examined for their effects on the Apo activation process. While SO2 had little effect, CS2 was shown to inhibit Apo activation via its ability to substitute for CO2 to yield an inactive dithiocarbamate-containing protein. Sulfur-to-Ni charge-transfer transitions arising from this species yielded an electronic absorption band at 324 nm with a shoulder at 382 nm. Borate, sulfate, phosphate, and molybdate had essentially no effect on Apo activation and did not substitute for bicarbonate, while treatment of Apo with Ni(II) plus vanadate led to the production of active urease containing two Ni and one V per active site. Vanadate-dependent activation of Apo resembled the normal activation process in terms of concentration of anion required, optimal pH, and incubation time needed. Furthermore, the UV-visible spectrum, maximal specific activity [386 +/- 26 U.(mg of protein)-1], Km (1.83 +/- 0.20 mM urea), and pH dependence for the vanadate-containing urease were essentially identical to properties observed for bicarbonate-activated enzyme. Vanadate-activated Apo is proposed to possess a vanadylated lysine that bridges the two Ni ions comprising its metallocenter. PMID- 9398240 TI - Characterization of alanine-rich peptides, Ac-(AAKAA)n-GY-NH2 (n = 1-4), using vibrational circular dichroism and Fourier transform infrared. Conformational determination and thermal unfolding. AB - Vibrational circular dichroism (VCD) and Fourier transform IR (FTIR) were measured for a series of short alanine-based peptides having the general formula Ac-(AAKAA)n-GY-NH2 (n = 1-4) from 5 to 50 degrees C in D2O and at room temperature in both TFE and H2O. In both of these latter solvents, the dominant structural form at the lowest temperature for the longest oligomers is alpha helical. The same is true for the n = 4 peptide in D2O, but under these more dilute aqueous conditions, the shorter (n = 3) peptides have mixed helix-coil structures and the n = 1 and 2 peptides are random coils. The VCD data do not support the 310-helix as a dominant contributor to the conformation of these oligomers in any of these solvents. These vibrational spectral data are consistent with lower-concentration electronic CD results and additionally indicate increased helical stability at higher concentrations. VCD amide I data for the 22mer (n = 4) in D2O indicate that the peptide undergoes a transition from a highly helical conformation at 5 degrees C to a dominant random coil structure at approximately 45 degrees C with a Tm of approximately 25 degrees C (effective midpoint). Factor analysis of the thermal data showed that three principal components were required to describe both the VCD and FTIR data for the n = 4 peptide in D2O. The transition is characterized by a gradual loss of contribution from a spectral component representing the alpha-helical fraction. The third component is evidence of an optically detected intermediate conformation best viewed as a mixed coil-helix structure resulting from end fraying of the helical peptide as the temperature is increased. The nature of the junction between the interior helix and frayed ends is not determined by these data and could involve local (phi and psi) angles mimicking a 310-helix that would provide consistency with ESR and NMR results from Millhauser and co workers. PMID- 9398241 TI - Effect of prolyl isomerase on the folding reactions of staphylococcal nuclease. AB - The low-temperature fluorescence-detected refolding of staphylococcal nuclease (SNase) can be described by three slow kinetic phases. The slowest phase is absent in the P117G mutant of SNase. Peptidyl prolyl cis-trans isomerase (cyclophilin), which has been shown to catalyze the slow folding reactions of some proteins, was employed to determine which of the refolding reactions of SNase and P117G SNase involve proline isomerization. We report here that all three folding phases of the wild type and the slower phase of P117G SNase are catalyzed by prolyl isomerase, indicating that proline isomerization is involved in these fluorescence-detected phases in the refolding of SNase. Since the rates of these phases are denaturant-dependent, we conclude that the slow folding steps involve isomerization of non-native cis proline peptide bonds and are tightly coupled to denaturant-sensitive structural changes. PMID- 9398242 TI - The conformation of the alpha-helical coiled coil domain of macrophage scavenger receptor is pH dependent. AB - Macrophage scavenger receptor is a trimerized membrane protein that binds ligands and undergoes internalization by endocytosis. The receptor releases the ligands in the endosome, and then is recycled. The mechanisms of the ligand release and the recycling of the receptor have not been clearly determined. We analyzed the structure of the alpha-helical coiled coil domain considered to be responsible for acid-mediated ligand dissociation, by chemical cross-linking, sedimentation equilibrium, Western blot, and circular dichroism analyses. This domain has 22 heptad repeats, which are characteristic of the sequence of an alpha-helical coiled coil structure, with a discontinuity in the middle. We prepared three peptides, corresponding to the entire alpha-helical coiled coil domain (alpha), its N-terminal half (alpha-N), and its C-terminal half (alpha-C), by expression of each gene in Escherichia coli. The alpha and alpha-N peptides show triple stranded alpha-helical coiled coil structures, but in contrast, the alpha-C peptide shows a random structure. When connected to the N-terminus by a chemical ligation method, the alpha-C peptide also shows an alpha-helical coiled coil structure, but only at an acidic pH. These results suggest that the N-terminus of the alpha-helical coiled coil domain is responsible for the formation of a stable trimer and the C-terminus exhibits the pH-dependent conformational change that might be involved in the ligand release by the macrophage scavenger receptor. PMID- 9398243 TI - Characterization of the reaction mechanism for Trypanosoma brucei ornithine decarboxylase by multiwavelength stopped-flow spectroscopy. AB - Ornithine decarboxylase (ODC), a pyridoxal 5'-phosphate (PLP)-dependent enzyme, catalyzes the first committed step in the biosynthesis of polyamines. The UV visible spectra of PLP (300-500 nm) was used to monitor the formation and breakdown of ODC reaction intermediates by multiwavelength stopped-flow spectroscopy to determine the reaction mechanism. Global kinetic analysis of the spectral data acquired after mixing ODC with saturating substrate (S) or product (P) (10 mM ornithine or 10 mM putrescine at 4 degrees C) suggests that ODC catalyzed decarboxylation proceeds by the following reaction mechanism: ODC + S if A --> B --> C --> D --> E/F if ODC + P, where A-F are intermediates along the reaction path. Species B, which has absorbance maxima of 350 and 450 nm, is spectrally distinct from the other intermediates. On the basis of the calculated spectral characteristics, species B is likely to represent a quinoid intermediate which would be formed directly upon decarboxylation of ornithine. Thus, the data suggest that the reaction proceeds via formation of a Schiff base intermediate (species A) during the dead time of the stopped-flow instrument, followed by formation of a quinoid intermediate with a rate constant of 21 s-1. The quinoid intermediate decays in two steps (with rates of 145 and 1.0 s-1, respectively) to a Schiff base with putrescine (species D). Protonation of the Calpha carbon is required for the formation of species D, suggesting that the first of these events represents this step. The decay of species D to free enzyme and product occurs via a minimum of two intermediates and at an overall rate constant of 1-3 s-1. By comparison to the steady-state turnover number (kcat = 0.5 s-1 at 4 degrees C), these data identify product release as a rate-determining step in the overall reaction. PMID- 9398244 TI - A biophysical study of integral membrane protein folding. AB - In order to characterize the thermodynamic constraints on the process of integral membrane protein folding and assembly, we have conducted a biophysical dissection of the structure of bacteriorhodopsin (BR), a prototypical alpha-helical integral membrane protein. Seven polypeptides were synthesized, corresponding to each of the seven transmembrane alpha-helices in BR, and the structure of each individual polypeptide was characterized in reconstituted phospholipid vesicles. Five of the seven polypeptides form stable transmembrane alpha-helices in isolation from the remainder of the tertiary structure of BR. However, using our reconstitution protocols, the polypeptide corresponding to the F helix in BR does not form any stable secondary structure in reconstituted vesicles, and the polypeptide corresponding to the G helix forms a hyperstable beta-sheet structure with its strands oriented perpendicular to the plane of the membrane. [The polypeptide corresponding to the C helix spontaneously equilibrates in a pH-dependent manner between a transmembrane alpha-helical conformation, a peripherally bound nonhelical conformation, and a fully water soluble conformation; the conformational properties of this polypeptide are the subject of the accompanying paper: Hunt et al. (1997) Biochemistry 36, 15177-15192.] Our observations suggest that the folding of alpha-helical integral membrane proteins may proceed spontaneously. However, the preference for a non-native conformation exhibited by two of the polypeptides suggests that the formation of some transmembrane substructures could require external constraints such as the links between the helices, interactions with the rest of the protein, or the involvement of cellular chaperones or translocases. Our results also suggest a strategy for improving the thermodynamic stability of alpha-helical integral membrane proteins, a goal that could facilitate attempts to overexpress and/or refold them. PMID- 9398245 TI - Spontaneous, pH-dependent membrane insertion of a transbilayer alpha-helix. AB - A question of fundamental importance concerning the biosynthesis of integral membrane proteins is whether transmembrane secondary structure can insert spontaneously into a lipid bilayer. It has proven to be difficult to address this issue experimentally because of the poor solubility in aqueous solution of peptides and proteins containing these extremely hydrophobic sequences. We have identified a system in which the kinetics and thermodynamics of alpha-helix insertion into lipid bilayers can be studied systematically and quantitatively using simple spectroscopic assays. Specifically, we have discovered that a 36 residue polypeptide containing the sequence of the C-helix of the integral membrane protein bacteriorhodopsin exhibits significant solubility in aqueous buffers free of both detergents and denaturants. This helix contains two aspartic acid residues in the membrane-spanning region. At neutral pH, the peptide associates with lipid bilayers in a nonhelical and presumably peripheral conformation. With a pKa of 6.0, the peptide inserts into the bilayer as a transbilayer alpha-helix. The insertion reaction proceeds rapidly at room temperature and is fully reversible. PMID- 9398246 TI - Interruption of G protein-coupling in CXCR2 does not alter ligand binding, but eliminates ligand-activation of GTPgamma35S binding, calcium mobilization, and chemotaxis. AB - CXCR2 is a seven-transmembrane receptor that transduces intracellular signals in response to the chemokines IL-8, MGSA/GRO, and other ELR motif-containing CXC chemokines by coupling to heterotrimeric GTP-binding proteins. In this study, we have mutated two putative G protein-coupling regions of CXCR2 and characterized the effects of these mutations on ligand-activated signal transductions: aspartic acid 89 in the second transmembrane domain and the HRAMR sequence (BBXXB motif, found in the third intracellular loop where B indicates a basic amino acid and X represents any amino acid). The Asp89 was replaced by either asparagine (D89N) or glutamic acid (D89E). For the BBXXB motif, the first two basic amino acids were mutated to two neutral isoleucines (HR-II), or alternatively, two isoleucines were inserted between alanine and methionine (II-insert). When expressed in human embryonic kidney 293 cells, the D89E mutant was localized intracellularly with no detectable cell surface expression. In contrast, D89N, HR-II, and II-insert mutants displayed cell surface expression, with Kd values and expression levels similar to that of the wild-type transfectant. The ability of the mutants to transduce signal was assessed by ligand-stimulated GTPgamma35S binding, mobilization of intracellular free Ca2+, and chemotaxis assays. Both D89N and HR II mutants signaled similarly to a wild-type receptor in all three assays. However, the II-insert mutant exhibited a loss of ligand-stimulated GTPgamma35S binding, calcium mobilization, and chemotaxis. Unexpectedly, this receptor underwent ligand-induced sequestration comparable to wild-type CXCR2. These data indicate that Asp89 and the basic amino acids in the third intracellular domain do not play essential roles in ligand-induced signal transduction through CXCR2. However, proper secondary structure and orientation of the third intracellular loop of CXCR2 are essential for ligand-mediated signal transduction but not for receptor sequestration. PMID- 9398247 TI - Oligomerization of a 45 kilodalton fragment of diphtheria toxin at pH 5.0 to a molecule of 20-24 subunits. AB - Diphtheria toxin (DT) is a 58 kDa protein, secreted by lysogenic strains of Corynebacterium diphtheriae, that causes the disease diphtheria in humans. The catalytic (C) domain of DT kills host cells by gaining entry into the cytoplasm and inhibiting protein synthesis. The translocation of the C domain across the endosomal membrane and into the cytoplasm of a host cell is mediated by the translocation (T) domain of DT. This process is triggered by acidification from pH approximately 7 to pH approximately 5 within the endosome. Here we show that crm45 (cross-reacting material of 45 kDa), a 45 kDa deletion mutant of DT which contains the C and T domains but lacks the C-terminal receptor-binding (R) domain, undergoes a transition from a monomer to a large oligomer upon acidification from pH 7.0 to pH 5.0. Dynamic light scattering analysis of crm45 at pH 5.0 results in a polydispersity value of only 8-17%, suggesting that the oligomer is uniformly sized. Using analytical ultracentrifugation, measurements of the sedimentation rate and diffusion coefficient of crm45 at pH 5.0 result in a molecular mass determination of 890 +/- 40 kDa (20 +/- 1 subunits) for the oligomer. Equilibrium sedimentation data on crm45 at pH 5.0 are best fit by a single species with a mass of 1000 +/- 50 kDa (24 +/- 1 subunits). These results reveal the pH-dependent formation of a uniformly sized, 20-24 subunit oligomer of the C and T domains of DT, in solution. Because the oligomer of crm45 forms at the pH of the acidified endosome, it could be relevant to the translocation of the C domain of DT across the endosomal membrane and into the cytoplasm of host cells. The possible relevance of this oligomer of crm45 to the membrane translocation of the C domain of DT correlates with earlier kinetic studies of DT intoxication of Vero cells, which inferred the transfer of approximately 20 C domains of DT to the cytoplasm of host cells, in a single event. PMID- 9398248 TI - Binding of steroid modulators to recombinant cytosolic domain from mouse P glycoprotein in close proximity to the ATP site. AB - We recently found that recombinant NBD1 cytosolic domain corresponding to segment 395-581 of mouse mdr1 P-glycoprotein bound fluorescent 2'(3')-N methylanthraniloyl-ATP (MANT-ATP) with high affinity [Dayan, G., Baubichon Cortay, H., Jault, J.-M., Cortay, J. -C., Deleage, G., & Di Pietro, A. (1996) J. Biol. Chem. 271, 11652-11658]. The present work shows that a longer 371-705 domain (extended-NBD1), including tryptophan-696 as an intrinsic probe, which bound MANT-ATP with identical affinity, also interacted with steroids known to modulate anticancer drug efflux from P-glycoprotein-positive multidrug-resistant cells. Progesterone, which is not transported, its hydrophobic derivatives medroxyprogesterone acetate and megestrol acetate, and Delta6-progesterone produced nearly a 50% saturating quenching of the domain intrinsic fluorescence, with dissociation constants ranging from 53 to 18 microM. The even more hydrophobic antiprogestin RU 486 produced a complete quenching of tryptophan-696 fluorescence, in contrast to more hydrophilic derivatives of progesterone containing hydroxyl groups at positions 11, 16, 17, and 21 and known to be transported, which produced very little quenching. A similar differential interaction was observed with full-length purified P-glycoprotein. The steroid binding region within extended-NBD1 appeared distinct from the nucleotide-binding site as the RU 486-induced quenching was neither prevented nor reversed by high ATP concentrations. In contrast, MANT-ATP binding was efficiently prevented or displaced by RU 486, suggesting that the hydrophobic MANT group of the bound nucleotide analogue overlaps, at least partially, the adjacent steroid-binding region revealed by RU 486. PMID- 9398249 TI - Nitric oxide inhibition of lipid peroxidation: kinetics of reaction with lipid peroxyl radicals and comparison with alpha-tocopherol. AB - The reaction between nitric oxide (*NO) and lipid peroxyl radicals (LOO*) has been proposed to account for the potent inhibitory properties of *NO toward lipid peroxidation processes; however, the mechanisms of this reaction, including kinetic parameters and nature of termination products, have not been defined. Here, the reaction between linoleate peroxyl radicals and *NO was examined using 2, 2'-azobis(2-amidinopropane) hydrochloride-dependent oxidation of linoleate. Addition of *NO (0.5-20 microM) to peroxidizing lipid led to cessation of oxygen uptake, which resumed at original rates when all *NO had been consumed. At high *NO concentrations (>3 microM), the time of inhibition (Tinh) of chain propagation became increasingly dependent on oxygen concentration, due to the competing reaction of oxygen with *NO. Kinetic analysis revealed that a simple radical-radical termination reaction (*NO:ROO* = 1:1) does not account for the inhibition of lipid oxidation by *NO, and at least two molecules of *NO are consumed per termination reaction. A mechanism is proposed whereby *NO first reacts with LOO* (k = 2 x 10(9) M-1 s-1) to form LOONO. Following decomposition of LOONO to LO* and *NO2, a second *NO is consumed via reaction with LO*, with the composite rate constant for this reaction being k = 7 x 10(4) M-1 s-1. At equal concentrations, greater inhibition of oxidation was observed with *NO than with alpha-tocopherol. Since *NO reacts with LOO* at an almost diffusion-limited rate, steady state concentrations of 30 nM *NO would effectively compete with endogenous alpha-tocopherol concentrations (about 20 microM) as a scavenger of LOO* in the lipid phase. This indicates that biological *NO concentrations (up to 2 microM) will significantly influence peroxidation reactions in vivo. PMID- 9398250 TI - Conformational changes in aerolysin during the transition from the water-soluble protoxin to the membrane channel. AB - Proteolytic activation, oligomerization, and membrane insertion are three steps that precede channel formation by the bacterial toxin aerolysin. Using attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) and hydrogen deuterium exchange, the structural changes associated with each step were analyzed. Our results show that activation induces a significant change in secondary structure, characterized by a decrease in random structure and an increase in beta-sheet content. We show that release of the propeptide is essential for this conformational change to occur and that changes are not restricted to the vicinity of the cleavage site but appear to propagate along the molecule. In contrast, subsequent oligomerization of the mature toxin does not involve any change in overall secondary structure but does involve a modification of the tertiary interactions. Finally, insertion of the heptameric complex into dimyristoylphosphatidylcholine vesicles also occurs without major modification of the secondary structure. Studies on the orientations of the secondary structures of the heptamer in the lipid bilayer have also been performed. PMID- 9398251 TI - Effects of clusterin overexpression on TNFalpha- and TGFbeta-mediated death of L929 cells. AB - Clusterin is a widely distributed and highly conserved protein for which many functions have been proposed. We used transfected L929 cells to study the effect of clusterin expression on the regulation of cell death signals. We showed that high levels of clusterin expression, about 0.2 pg clusterin secreted per cell per 48 h period, specifically protected L929 cells from TNFalpha-mediated cytotoxicity, while low expression (about 4 fg/cell/48 h) had no effect. However, clusterin expression did not provide transfected L929 cells with protection against death mediated by colchicine, staurosporine or azide. High level expression of clusterin in transfected L929 cells also potentiated the cytotoxicity of TGFbeta. It had previously been shown that exposure of L929 cells to TGFbeta provides protection against TNFalpha. We showed that this protective effect is not additive to that of clusterin expression. One interpretation of this data is that it suggests that clusterin and TGFbeta may act via a common mechanism to provide protection against the cytotoxicity of TNFalpha. Our results indicate that an intracellular action of clusterin protein is responsible for protection against TNFalpha cytotoxicity. Exposure to TNFalpha induces an increase in the level of cell-associated clusterin and specifically in the level of a novel clusterin molecule, which when analyzed under reducing conditions by SDS/PAGE and immunoblotting appears as two closely spaced bands at about 36 and 38.5 kDa. When analyzed under the same conditions, the normal form of intracellular clusterin, which is present with or without exposure to TNFalpha, appears as two poorly resolved bands at about 43-45 kDa. Since the novel form of clusterin is also expressed in cells exposed to TGFbeta, colchicine, staurosporine, and azide, it may result from toxin-induced disruption of processes of normal cellular protein production. PMID- 9398252 TI - Photoassembly of the photosystem II (Mn)4 cluster in site-directed mutants impaired in the binding of the manganese-stabilizing protein. AB - Photoactivation is the light-dependent ligation of Mn2+ into the H2O oxidation complex of photosystem II (PSII) and culminates in the formation of an enzymatically active complex containing Ca2+ and four Mn>/=3+. Previous kinetic analysis demonstrated that the genetic removal of the extrinsic manganese stabilizing protein (MSP) increases the quantum yield of photoactivation 4-fold relative to that of the wild type, consistent with the hypothesis that MSP hinders access of Mn2+ to a site of photoligation [Burnap, R. L., et al. (1996) Biochemistry35, 874-882]. In this report, several Synechocystis sp. PCC6803 mutants with defined amino acid substitutions in the N-terminal region of MSP or the e-loop of intrinsic PSII protein CP47 [Putnam-Evans, C., et al. (1996) Biochemistry 35, 4046-4053] were characterized in terms of the binding of MSP to the intrinsic portion of the PSII complex and in terms of photoactivation kinetics. The charge-pair switch mutation, Arg384Arg385 --> Glu384Glu385 in the lumenal e-loop of CP47 (CP47 RR384385EE), exhibited the most severe impairment of MSP binding, whereas the Arg384Arg385 --> Gly384Gly385 (CP47 RR384385GG) mutation caused a more moderate impairment in binding. Single-substitution mutations at the highly conserved Asp9 or Asp10 positions in the amino-terminal region of MSP also resulted in a reduced binding affinity, but not as severe as that in CP47 RR384385EE. The relative quantum yield of photoactivation of hydroxylamine extracted mutant PSII was generally found to correlate with the degree of MSP binding impairment, with the CP47 RR384385 mutants exhibiting the highest quantum yields. A two-locus, double-mutant construct involving deletion of MSP in the CP47 RR384385EE background was found to be only slightly more impaired in H2O oxidation activity than either of the corresponding single-locus mutant derivatives, indicating that mutations at these genetically separate loci encode physically interacting products affecting the same reaction parameter during H2O oxidation. Taken together, the results reinforce the concept that MSP interacts with the e-loop of CP47 at Arg384Arg385 and that disruption of this interaction causes significant alterations of the site of H2O oxidation in terms of assembly and enzymatic activity of the Mn cluster. PMID- 9398253 TI - Design of fluorescent substrates and potent inhibitors of CYP73As, P450s that catalyze 4-hydroxylation of cinnamic acid in higher plants. AB - CYP73As are the major functional cytochromes P450 in higher plants. Several of them have been shown to catalyze the 4-hydroxylation of cinnamic acid, the first oxidative step in the synthesis of lignin, flavonoids, coumarins, and other phenylpropanoids. The coding sequence for CYP73A1, the enzyme from Helianthus tuberosus, has been isolated and expressed in yeast. Previous studies indicate that the yeast-expressed enzyme is capable of metabolizing cinnamic acid and several small, planar molecules but with low efficiency. Using this we further examined how CYP73A1 could bind and metabolize a set of possible alternate substrates. We show here that naphthalenes, quinolines, and indoles substituted with an aldehyde, a carboxylic, or a sulfonic acid group make good ligands and substrates for CYP73A1. The best ligands are hydroxynaphthoic acids, which show higher affinity than cinnamate. Naphthalene, 2-naphthol, and molecules with two carbon side chains, such as natural and synthetic auxins, are not substrates of this enzyme. Methyl-2-naphthoate and 2-hydroxy-1-naphthoic acid are strong ligands of CYP73A1 but are not metabolized. Uncoupling and low spin conversion induced by these compounds suggest that their positioning in the heme pocket is inadequate for catalysis. These compounds can act as potent inhibitors of the second step of the phenylpropanoid pathway, the first described so far. The molecule which most closely mimics cinnamic acid, 2-naphthoic acid, is metabolized with a catalytic turnover and efficiency similar to those measured with the physiological substrate. Using this compound we designed a fluorometric assay to measure the catalytic activity of CYP73As. This assay was then used to monitor the CYP73As activity in microsomes from transgenic yeast and several plant species. PMID- 9398254 TI - Energy transfer in LHCII monomers at 77K studied by sub-picosecond transient absorption spectroscopy. AB - Energy transfer from chlorophyll b (Chl b) to chlorophyll a (Chl a) in monomeric preparations of light-harvesting complex II (LHCII) from spinach was studied at 77 K using pump-probe experiments. Sub-picosecond excitation pulses centered at 650 nm were used to excite preferentially Chl b and difference absorption spectra were detected from 630 to 700 nm. Two distinct Chl b to Chl a transfer times, approximately 200 fs and 3 ps, were found. A clearly distinguishable energy transfer process between Chl a molecules occurred with a time constant of 18 ps. The LHCII monomer data are compared to previously obtained LHCII trimer data, and both data sets are fitted simultaneously using a global analysis fitting routine. Both sets could be described with the following time constants: 140 fs, 600 fs, 8 ps, 20 ps, and 2.9 ns. In both monomers and trimers 50% of the Chl b to Chl a transfer is ultrafast (<200 fs). However, for monomers this transfer occurs to Chl a molecules that absorb significantly more toward shorter wavelengths than for trimers. Part of the transfer from Chl b to Chl a that occurs with a time constant of 600 fs in trimers is slowed down to several picoseconds in monomers. However, it is argued that observed differences between monomers and trimers should be ascribed to the loss of some Chl a upon monomerization or a shift of the absorption maximum of one or several Chl a molecules. It is concluded that Chl b to Chl a transfer occurs only within monomeric subunits of the trimers and not between different subunits. PMID- 9398255 TI - Conformation-activated protonation in reaction centers of the photosynthetic bacterium Rhodobacter sphaeroides. AB - Kinetics and stoichiometry of proton binding/unbinding induced by intense (1 W cm 2) and continuous illumination were measured in the isolated reaction center (RC) protein from photosynthetic purple bacterium Rhodobacter sphaeroides in the absence of an external electron donor. At high ionic strength (100 mM), large proton release (approximately 6 H+ per RC) was observed at pH 6 and substoichiometric H+-ion binding (approximately 0.3 H+ per RC) at pH 8. These observations together with optical spectroscopy on the oxidized dimer indicate that, at room temperature, two distinct conformations of the RC can be obtained depending on the pH, Eh, and illumination. Acidic pH, a large redox gap between the actual Eh of the solution and the midpoint potential of the acceptor quinone, and strong illumination favor the conversion of the RC from the dark-adapted state to the light-adapted state. These conformations differ greatly in the rates of primary photochemistry, the reoxidation of semiquinone and the rereduction of the oxidized dimer, and the protonation states of the amino acids of the protein. Whereas substoichiometric proton unbinding is observed in the P+Q redox state of the protein in the dark-adapted conformation, much larger H+-ion release is detected in the light-adapted conformation. From the pH dependence of the key processes in the conformational change and reoxidation of semiquinone, we concluded that they are controlled by protonatable groups available in the protein. A simple phenomenological model is presented that relates the rates and equilibrium constants of the electron transfer reactions and the conformational change of the RC. PMID- 9398256 TI - Involvement of the reductase domain of neuronal nitric oxide synthase in superoxide anion production. AB - Neuronal nitric oxide synthase (nNOS) is a modular enzyme which consists of a flavin-containing reductase domain and a heme-containing oxygenase domain, linked by a stretch of amino acids which contains a calmodulin (CaM) binding site. CaM binding to nNOS facilitates the transfer of NADPH-derived electrons from the reductase domain to the oxygenase domain, resulting in the conversion of L arginine to L-citrulline with the concomitant formation of a guanylate cyclase activating factor, putatively nitric oxide. Numerous studies have established that peroxynitrite-derived nitrogen oxides are present following nNOS turnover. Since peroxynitrite is formed by the diffusion-limited reaction between the two radical species, nitric oxide and O2.-, we employed the adrenochrome assay to examine whether nNOS was capable of producing O2.- during catalytic turnover in the presence of L-arginine. To differentiate between the role played by the reductase domain and that of the oxygenase domain in O2.- production, we compared its production by nNOS against that of a nNOS mutant (CYS-331), which was unable to transfer NADPH-derived electrons efficiently to the heme iron under special conditions, and against that of a flavoprotein module construct of nNOS. We report that O2.- production by nNOS and the CYS-331 mutant is CaM-dependent and that O2.- production can be modulated by substrates and inhibitors of nNOS. O2.- was also produced by the reductase domain of nNOS; however, it did not display the same CaM dependency. We conclude that both the reductase and oxygenase domains of nNOS produce O2.-, but that the reductase domain is both necessary and sufficient for O2.- production. PMID- 9398257 TI - Cross-linking and N-(1-pyrenyl)maleimide labeling of cysteine mutants of proton pumping pyridine nucleotide transhydrogenase of Escherichia coli. AB - The pyridine nucleotide transhydrogenase of Escherichiacoli is a proton pump composed of two subunits (alpha and beta) organized as an alpha2beta2 tetramer. The enzyme contains seven cysteine residues, five in the alpha-subunit and two in the beta-subunit. The reaction of these residues with the cross-linking agent cupric 1, 10-phenanthrolinate and with the fluorescent thiol reagent N-(1 pyrenyl)maleimide was investigated in mutants in which one or more of these cysteine residues had been mutated to serine or threonine residues. Mutation of alphaCys395 and alphaCys397 prevented disulfide bond formation to give the cross linked alpha2 dimer. We concluded that the two alpha-subunits of the holoenzyme interface in the region of these two cysteine residues. Pyrenylmaleimide reacted with detergent-washed cytoplasmic membrane vesicles containing high levels of transhydrogenase protein to show characteristic fluorescence emission bands at 378-379, 397-398, and 419-420 nm. At higher ratios of pyrenylmaleimide:transhydrogenase (>5:1) and longer times of reaction, an eximer band at 470 nm was formed. This was attributed to interaction between noncovalently bound molecules of pyrenylmaleimide. The cysteine residues of the beta-subunit (betaCys147 and betaCys260) were covalently modified by pyrenylmaleimide. betaCys147 reacted more strongly than betaCys260 with the fluorophore, and the pyrene derivative of betaCys147 was more accessible to quenching by 5-doxylstearate, suggesting a proximity to the surface of the membrane. Covalent modification of betaCys260 resulted in inhibition of enzyme activity. The inhibition was attributed to the introduction of the bulky pyrene group into the enzyme. PMID- 9398258 TI - Charge recombination and proton transfer in manganese-depleted photosystem II. AB - The proton transfer reactions induced by the oxidation and reduction of the secondary donor, tyrosine YZ, have been studied in photosystem II after inactivation (Mn-depletion) of the oxygen-evolving complex. The rate of the recombination reaction of YZox with the reduced primary acceptor QA- appears modulated by a protonatable group with pK approximately 6 in the presence of YZox. The finding of monophasic recombination kinetics requires that the proton equilibration of this group is faster than the recombination rate. The same group modulates the extent of proton release, from 0 below pH 5 to 1 per center above pH 7. The kinetics of proton appearance and disappearance in the bulk medium are markedly dependent on the material used. In PSII core particles, the release is observed in the 100 micros range and the uptake accompanies the recombination reaction. In PSII membranes, both of these reactions are markedly delayed, so that the uptake considerably lags behind the completion of the recombination reaction. An electrochromic shift of a chlorophyll is present during the whole lifetime of YZox, suggesting a charged character of this species. A fast decreasing phase of this signal was observed in particles in the same time range as proton release. These results are discussed in the framework of a model where the proton originating from the formation of the neutral oxidized tyrosine radical (YZ.) remains locally trapped. In turn, this proton shifts the pK of a nearby group from a value >/=9 to a value of 6. PMID- 9398259 TI - A Pneumocystis carinii group I intron ribozyme that does not require 2' OH groups on its 5' exon mimic for binding to the catalytic core. AB - The recent increase in the population of immunocompromised patients has led to an insurgence of opportunistic human fungal infections. The lack of effective treatments against some of these pathogens makes it important to develop new therapeutic strategies. One such strategy is to target key RNAs with antisense compounds. We report the development of a model system for studying the potential for antisense targeting of group I self-splicing introns in fungal pathogens. The group I intron from the large ribosomal subunit RNA of mouse-derived Pneumocystis carinii has been isolated and characterized. This intron self-splices in vitro. A catalytically active ribozyme, P-8/4x, has been constructed from this intron to allow measurement of dissociation constants for potential antisense agents. At 37 degrees C, in 50 mM Hepes (25 mM Na+), 15 mM MgCl2, and 135 mM KCl at pH 7.5, the exogenous 5' exon mimic r(AUGACU) binds about 60 000 times more tightly to this ribozyme than to r(GGUCAU), a mimic of its complementary binding site on the ribozyme. This enhanced binding is due to tertiary interactions. This tertiary stabilization is increased by single deoxynucleotide substitutions in the exon mimic at every position except for the internal A, which is essentially unchanged. Thus 2' OH groups of the 5' exon mimic do not form stabilizing tertiary interactions with the P-8/4x ribozyme, in contrast to the Tetrahymena L 21 ScaI ribozyme. Furthermore, at 37 degrees C, the exogenous 5' exon mimic d(ATGACT) binds nearly 32 000 times more tightly to the P-8/4x ribozyme than to r(GGUCAU). Therefore, oligonucleotides without 2' OH groups can exploit tertiary stabilization to bind dramatically more tightly and with more specificity than possible from base pairing. These results suggest a new paradigm for antisense targeting: targeting the tertiary interactions of structural RNAs with short antisense oligonucleotides. PMID- 9398260 TI - A new type of DNA minor-groove complex: carbazole dication-DNA interactions. AB - The effect of opportunistic infections (OI) on immune-compromised populations has been known for decades, but the recent AIDS epidemic has sparked renewed interest in the development of new anti-OI agents. The mechanism of action of a series of cationic unfused-aromatic anti-OI drugs is believed to involve binding of the drug to AT sequences in the minor groove of DNA. Some new anti-OI drug candidates have been synthesized with fused aromatic ring systems (e.g. carbazoles) that do not resemble the classical paradigm for minor-groove interactions at AT sequences in DNA. To characterize the DNA interactions of these compounds, we have used UV vis absorbance, fluorescence, kinetic measurements, and circular dichroism in conjunction with NMR spectroscopy to evaluate the structure of the complexes formed between the carbazoles and DNA. Application of these methods to carbazoles substituted at either the 3,6 or 2,7 positions with cationic imidazoline groups gave conclusive, but very surprising, evidence that both compounds bind strongly in the minor groove at AT DNA sequences. NMR and molecular modeling of the complexes formed between the 3,6- and 2,7-carbazoles and the self-complementary oligomer d(GCGAATTCGC) have been used to establish structural details for the minor-groove complex. These results have been used as constraints for molecular modeling calculations to construct models of the minor-groove-carbazole complexes and to draw conclusions regarding the molecular basis for the effects of substituent position on carbazole-DNA affinities. The surprising result is that the 2,7 carbazole binds in AT sequences with hydrogen bonds involving one imidazoline group and the carbazole NH. The 3,6-carbazole compound binds in a more "classical" model that uses both imidazoline groups for H-bonding while the carbazole NH points out of the minor groove. The carbazoles thus form a new type of DNA minor groove complex and their excellent biological activities indicate that a variety of fused-ring minor-groove binding agents should be investigated. PMID- 9398261 TI - The relative stabilities of base pair stacking interactions and single mismatches in long RNA measured by temperature gradient gel electrophoresis. AB - The thermal stability of RNA duplexes differing by a single base pair (bp) substitution or mismatch were investigated by temperature gradient gel electrophoresis (TGGE). All base pair substitutions and mismatches were examined at six sites, and limited changes were investigated at three other sites. DNA templates for in vitro transcription were generated by the polymerase chain reaction (PCR). Transcribed forward and reverse single stranded RNAs were annealed to form 345 bp dupex RNA. Solution melting curves of selected RNAs were in good agreement with the predicted three step transitions. Parallel TGGE was used to determine the relative stabilities of the RNAs, and perpendicular TGGE was employed to obtain mobility transitions and midpoint transition temperatures (Tmu) of the RNAs' first melting domain. The gel solvent included formamide and urea. The Tmu values of the first melting domain were influenced by the identity of the base pair substitution or mismatch as well as by the site's neighboring base pairs. The difference in the transition temperatures (deltaTmu) between pairs of RNA ranged from 0 to 5 degrees C. deltaTmu values were used to determine free energy differences (deltaDeltaG). For RNA pairs distinguished by a base pair substitution, the deltaDeltaG values were closely correlated with free energy differences calculated from stacking free energies determined from melting studies in 1 M Na+ [Serra, M. J., and Turner, D. H. (1995) Methods Enzymol. 259, 242-261.] An algorithm was developed using the free energies of terminal mismatches [Serra, M. J., and Turner, D. H. (1995) Methods Enzymol. 259, 242-261] that provided very good agreement with experimental free energies for the single internal mismatches. PMID- 9398262 TI - Kinetics of DNA polymerase I (Klenow fragment exo-) activity on damaged DNA templates: effect of proximal and distal template damage on DNA synthesis. AB - Mutagenic DNA adducts have been analyzed with respect to the rate of nucleotide insertion opposite the modified base, extension from that "mispair", and nucleotide insertion preference. To complement and extend these studies we have investigated the long-range effects of DNA adducts on DNA polymerase activity. To address this question, primer extension reactions were performed using DNA polymerase I, Klenow fragment exo-. Templates containing 7,8-dihydro-8 oxoguanine, dG-C8-aminofluorene, dG-C8-(acetylamino)fluorene, and the model abasic site, tetrahydrofuran, were used for these studies, and the steady-state kinetics of correct nucleotide insertion were determined at positions (-2), (-1), (+1), (+2), (+3), and (+5) with respect to the template lesion. The kinetics of primer extension by Klenow fragment exo- at template positions 3' to the lesion showed only a small inhibitory effect, <3-fold, even for the strongly blocking lesion, dG-C8-(acetylamino)fluorene, indicating that Klenow fragment exo- activity is not greatly affected by lesions in the single-stranded portion of the template-primer. In contrast, a dramatic decrease in the frequency of primer extension was observed at template sites 5' to the site of adduction. Inhibition of polymerase activity decreased as the distance from the lesion increased; however, a relatively large effect was seen at the (+2) and (+3) positions for dG C8-(acetylamino)fluorene and tetrahydrofuran. For these blocking lesions, the effect on extension 5 bases from the lesion was greatly reduced. We conclude from these studies that DNA damage at positions remote from the site of the lesion affects DNA polymerase function. PMID- 9398263 TI - The cyanobacterial repressor SmtB is predominantly a dimer and binds two Zn2+ ions per subunit. AB - The Synechococcus PCC7942 metallothionein repressor gene smtB has been cloned into a high expression vector and the protein purified to near homogeneity (>/=98%). Analytical ultracentrifugation studies demonstrate that the protein is predominantly dimeric in 0.1 M NaCl, pH 7.4, and 22 degrees C, exhibiting a monomer-dimer-tetramer equilibrium. The monomer-dimer (Ka(1,2)) and the dimer tetramer (Ka(2,4)) association constants are 3.24 x 10(5) and 9.90 x 10(2) M-1, respectively. The repressor binds two Zn2+ ions per subunit with an overall Kd of 3.49 x 10(-6) M. In the presence of Zn2+, Ka(1, 2) increases by 2 orders of magnitude to 1.25 x 10(7) M-1 and the apparent weight-averaged sedimentation coefficient increases from 2. 00 to 2.22 S. The fact that the increase in sedimentation coefficient is greater than that predicted by increased dimerization is interpreted as caused by compaction of the structure in the presence of metal ions. At pH 6.0, 0.1 M NaCl, and 22 degrees C, the protein exhibits only a monomer-dimer equilibrium, with Ka(1,2) = 1.52 x 10(7) M-1 which is almost identical to that seen upon binding Zn2+ at pH 7.4. The compaction and conformational change in SmtB caused by Zn2+ is consistent with a role for this altered quaternary state in derepression of smtA in Synechococcus challenged with heavy metal ions. PMID- 9398264 TI - Mechanism of DNA binding enhancement by hepatitis B virus protein pX. AB - At least three hundred million people worldwide are infected with the hepatitis B virus (HBV), and epidemiological studies show a clear correlation between chronic HBV infection and the development of hepatocellular carcinoma. HBV encodes a protein, pX, which abducts the cellular transcriptional machinery in several ways including direct interactions with bZIP transcription factors. These interactions increase the DNA affinities of target bZIP proteins in a DNA sequence-dependent manner. Here we use a series of bZIP peptide models to explore the mechanism by which pX interacts with bZIP proteins. Our results suggest that pX increases bZIP.DNA stability by increasing the stability of the bZIP dimer as well as the affinity of the dimer for DNA. Additional experiments provide evidence for a mechanism in which pX recognizes the composite structure of the peptide.DNA complex, not simply the primary peptide sequence. These experiments provide a framework for understanding how pX alters the patterns of transcription within the nucleus. The similarities between the mechanism proposed for pX and the mechanism previously proposed for the human T-cell leukemia virus protein Tax are discussed. PMID- 9398265 TI - Biochemical and mutational investigations of the enzymatic activity of macrophage migration inhibitory factor. AB - The protein mediator MIF has been identified as being released from immune cells by glucocorticoid stimulation and to counter-regulate glucocorticoid action. MIF also has been described recently to exhibit dopachrome tautomerase activity and to be structurally homologous to the bacterial enzymes 4-oxalocrotonate tautomerase (4-OT) and 5-carboxymethyl-2-hydroxymuconate isomerase (CHMI). We performed site-directed mutagenesis and biochemical analyses of mouse MIF in order to identify amino acid residues and protein domains that are essential for enzymatic reactivity. Mutant proteins which lacked a free N-terminal proline residue were enzymatically inactive, as was a preparation of native MIF modified covalently at its N terminus by 3-bromopyruvate, suggesting that this proline has a catalytic function. Substitutions of the internal histidine residues 42 and 63 did not affect enzymatic activity, indicating that these basic residues are not involved in dopachrome tautomerization. Carboxy-truncated forms of MIF (residues 1-110 and 1-104) also were inactive, affirming the role of the carboxy terminus in stable trimer formation and the importance of the trimer for enzymatic activity. Additional evidence for the homotrimeric structure of MIF under native solution conditions was obtained by SDS-PAGE analysis of MIF after chemical cross linking at low protein concentrations. The enzymatic activity of MIF was found to be reversibly inhibited by micromolar concentrations of fatty acids with chain lengths of at least 16 carbon atoms. Of note, molecular modeling of the substrate L-dopachrome methyl ester into the active site of MIF suggests an acid-catalyzed enzymatic mechanism that is different from that deduced from studies of the enzymes 4-OT and CHMI. Finally, in vitro analysis of an enzymatically inactive MIF species (P2 --> S) indicates that the glucocorticoid counter-regulatory activity of MIF can be functionally dissociated from its tautomerization activity. PMID- 9398266 TI - Activation of the 43 kDa inositol polyphosphate 5-phosphatase by 14-3-3zeta. AB - The 43 kDa inositol polyphosphate 5-phosphatase (5-phosphatase) hydrolyzes and thereby inactivates the second messenger molecules inositol 1,4,5-trisphosphate Ins(1,4,5)P3- and inositol 1,3,4,5-tetrakisphosphate in a signal terminating reaction. Recent studies have shown that the platelet protein pleckstrin forms a complex with the 43 kDa 5-phosphatase and activates Ins(1,4,5)P3 hydrolysis 2 fold [Auethavekiat, V., Abrams, C. S., & Majerus, P. W. (1997) J. Biol. Chem. 272, 1786-1790]. We now show that another platelet protein, 14-3-3zeta, forms a complex with the 43 kDa 5-phosphatase and thereby activates the hydrolysis of Ins(1,4,5)P3. Both pleckstrin and 14-3-3zeta contain one or more pleckstrin homology domains, both are present in platelet cytosol, and both dimerize and form complexes with other signalling proteins. Purified platelet pleckstrin and 14-3-3zeta enhanced the rate of the hydrolysis of Ins(1,4,5)P3 by the 43 kDa 5 phosphatase 1.9- and 3.8-fold, respectively, but did not activate the 75 kDa 5 phosphatase. We have demonstrated that the mechanism of 5-phosphatase activation by 14-3-3zeta results from specific complex formation between the 43 kDa 5 phosphatase and 14-3-3zeta. Recombinant 43 kDa 5-phosphatase bound to recombinant glutathione S-transferase (GST)/14-3-3zeta fusion protein, but not GST alone, immobilized on glutathione-Sepharose. A potential 14-3-3 binding motif was located in the 43 kDa, but not the 75 kDa, 5-phosphatase. The motif "363RSESEE" is present in close proximity to the proposed catalytic domain of the 43 kDa 5 phosphatase. A synthetic peptide corresponding to the putative 14-3-3 binding motif demonstrated specific, saturable binding to purified 125I-14-3-3, with a Kd of 92 nM. In addition, platelet cytosolic 5-phosphatase bound to recombinant 14-3 3zeta immobilized on glutathione-Sepharose. Thus, 14-3-3zeta serves in human platelets to activate the 43 kDa 5-phosphatase and may thereby function to prevent generation of Ins(1,4,5)P3 -mediated calcium release in unstimulated platelets. PMID- 9398267 TI - Cloning and characterization of cDNAs encoding chicken mitogen-activated protein kinase kinase type 2, MEK2: downregulation of MEK2 in response to inhibition of mitochondrial DNA expression. AB - The present work was initiated with the aim of identifying nuclear genes whose expression is sensitive to the mitochondrial DNA (mtDNA) status of transformed chicken DU24 cells. We cloned and sequenced cDNAs for the mitogen-activated protein kinase kinase type 2, MEK2, a protein involved in the mitogenic growth factor signal transduction pathway in vertebrates. Sequence comparisons between the chicken protein and its mammalian counterparts indicated that MEK2 proteins are highly conserved among vertebrates. Southern blot analysis of endonuclease digested genomic DNA from primary chick embryo fibroblasts (CEF) suggested that MEK2 is a single-copy gene in this vertebrate species. The steady-state level of MEK2 transcripts is decreased in DUS3 mtDNA-less (rho0) cells developed by long term exposure of DU24 rho+ cells to ethidium bromide (EtdBr). Run-on in vitro transcription assays and mRNA stability studies indicated that the decrease in MEK2 mRNA content is associated with post-transcriptional regulation. In parental DU24 cells, MEK2 mRNA content decreased after inhibition of mtDNA transcription by EtdBr and inhibition of translation on mitoribosomes by chloramphenicol (CAM). Cytoplasmic hybrids (cybrids) constructed by fusion of chicken rho0 cells with enucleated parental cells and CEF recovered a basal level of MEK2 expression. The MEK2 protein content is decreased in DUS3 rho0 cells and in parental DU24 rho+ cells treated with EtdBr and CAM for 6 days, while that of MEK1, a closely related kinase, remained unchanged. On the basis of these observations, we propose that mitochondria participate in the mitogenic signal transduction pathway in chicken cells through regulation of MEK2 expression. PMID- 9398268 TI - Demonstration of an extracellular ATP-binding site in NCAM: functional implications of nucleotide binding. AB - A minor fraction of the total ecto-type (E-type) ATPase activity of rat synaptosomes has been detected in immunoprecipitates of the neural cell adhesion molecule, NCAM, indicating that this either is an intrinsic enzymatic activity of NCAM or of an ATPase tightly associated to NCAM [Dzhandzhugazyan & Bock (1993) FEBS Lett. 336, 279-283]. We here demonstrate ATPase activity in preparations of the lipid-anchored as well as the transmembrane NCAM isoforms immunoisolated from transfected L-cells. A fraction of the E-type ATPase activity is spontaneously released from synaptosomes. Released material was fractionated by various chromatographic procedures and an extracellular fragment of NCAM was shown to co elute with the major part of the enzymatic activity. Furthermore, it was shown that agarose-coupled NCAM-antibodies retained 85% of the ATPase activity released from synaptosomes after treatment with phosphatidylinositol-specific phospholipase C. These findings restricted the association or expression of the enzymatic activity to the extracellular part of NCAM. An affinity reagent, 5'-p fluorosulfonylbenzoyl adenosine, FSBA, was shown to inhibit ATPase activity of immunoisolated NCAM, and incorporation of FSBA was detected in all three major NCAM isoforms (A, B, and C). An excess of ATP prevented both inactivation of the enzyme and affinity labeling of NCAM. Thus, NCAM contains an ATP-binding site, and this site is localized extracellularly and probably has the catalytic function. Binding of the substrate or FSBA protected a proteolytic cleavage site in NCAM localized close to the membrane presumably by induction of a local conformational change in NCAM, indicating a mechanism by which ATP may regulate NCAM adhesion and adhesion-triggered processes. A possible role of this mechanism in synaptic plasticity and memory consolidation is proposed. PMID- 9398269 TI - Mutations in the transmembrane domain of APP altering gamma-secretase specificity. AB - Alzheimer's disease (AD) beta-amyloid peptide (Abeta and betaA4) is derived from the amyloid precursor protein (APP) by the subsequent action of the so-far unidentified beta- and gamma-secretases. gamma-secretase, which generates the C terminus of Abeta, cleaves within the transmembrane domain of APP, preferentially after Abeta-residue 40 (Abeta 40) but also after residue 42 (Abeta 42). This Abeta 42 represents the major subunit of the plaques in AD. Since the position of gamma-secretase cleavage is crucial for understanding the pathogenic pathway, we investigated the effect of different point mutations at Thr43 on gamma-secretase specificity in SPA4CT (SPC99)-expressing COS7 cells. These constructs only require gamma-cleavage for Abeta release. We observed that all Thr43 mutations altered the specificity of gamma-secretase. Small hydrophobic residues favored the generation of Abeta 42, leading to an increase in the 42/40 ratio of Abeta (1.6-2.8-fold). The increase was even stronger (5.6-5.8-fold) when combined with the familial mutation Val46Phe. Thus, these constructs might be highly valuable for the generation of animal models for AD. Processing of full-length APP or SPA4CT yielded the same 42/40 ratio of Abeta (4. 7%). Both constructs, bearing the familial AD mutation Val46Phe, led to a similar increase in the 42/40 ratio (3.3- versus 3.6-fold). The p3 fragment, produced by alpha- and gamma-secretase, showed 42/40 ratios similar to Abeta when derived from wild-type and mutant proteins. These results suggest that the different Abeta- and p3-species are generated by gamma-cleavage activities with a similar enzymatic mechanism. PMID- 9398270 TI - An all-D amino acid peptide model of alpha1(IV)531-543 from type IV collagen binds the alpha3beta1 integrin and mediates tumor cell adhesion, spreading, and motility. AB - Type IV collagen promotes integrin-mediated cell adhesion, spreading, and motility. Several regions within the triple-helical domain of type IV collagen have been identified as tumor cellular recognition sites. Among these regions, the alpha1(IV)531-543 sequence, designated L-Hep-III, promotes integrin-mediated tumor cell adhesion and directly binds to the alpha3beta1 integrin [Miles, A. J., et al. (1994) J. Biol. Chem. 269, 30939-30945; Miles, A. J., et al. (1995) J. Biol. Chem. 270, 29047-29050]. We have presently compared the activities of the all-d enantiomeric peptide model of alpha1(IV)531-543, designated D-Hep-III, with L-Hep-III, for promoting the adhesion, spreading, and motility of metastatic melanoma and breast carcinoma cells. D-Hep-III was found to support melanoma and breast carcinoma cell adhesion, spreading, and motility in a dose-dependent fashion similar to that of L-Hep-III. The adhesions of melanoma and breast carcinoma cells to both type IV collagen and fibronectin were effectively inhibited by L-Hep-III and D-Hep-III. Melanoma cell invasion of the basement membrane was also inhibited by D-Hep-III. Characterization of the cell surface receptor for D-Hep-III was acheived via cell adhesion assays and affinity chromatography using monoclonal antibodies against integrin subunits. Immunoprecipitation analysis following EDTA elution from a D-Hep-III affinity column indicated that D-Hep-III binds to the alpha3beta1 integrin but not to the alpha2 or alpha6 integrin subunits. In summary, these studies demonstrate that an all-D model of the alpha1(IV)531-543 sequence mimics the biological activities of the all-L peptide. D-Hep-III is the first all-D peptide that has been shown to promote tumor cell adhesion, spreading, and migration, inhibit tumor cell adhesion and migration on type IV collagen and invasion of the basement membrane, and bind directly to an integrin. Due to the resistance to proteolysis, all-D receptor-binding peptides such as D-Hep-III have great potential for in vivo studies and as therapeutic agents. PMID- 9398271 TI - Evidence for a common active site for cleavage of an AP site and the benzene derived exocyclic adduct, 3,N4-benzetheno-dC, in the major human AP endonuclease. AB - We have previously reported that the 3,N4-benzetheno-dC (p-BQ-dC) endonuclease activity found in HeLa cells is a novel function of the major human AP endonuclease (HAP1) [Hang et al. (1996) Proc. Natl. Acad. Sci. U.S.A. 93, 13737 13741]. In this study, we compare the enzymatic and biochemical properties of the enzyme toward p-BQ-dC and an AP site in a defined oligonucleotide. A comparative analysis of the specificity constants (Kcat./Km) for p-BQ-dC and an AP site indicates that the AP site is the preferred substrate. The enzyme does not cleave other structurally related exocyclic adducts and modified nucleosides such as 1,N6-etheno-dA, 3,N4-etheno-dC, 1, N2-etheno-dG, 1,N2-propano-dG, 8-oxo-dG, and thymine glycol. The p-BQ-dC activity requires a double-stranded DNA substrate and is affected by the base in the opposite strand, with maximal activity for a p-BQ dC.G pair and minimal activity for a p-BQ-dC.C pair. The p-BQ-dC activity also requires Mg2+, Mn2+, or Zn2+ with optimal concentration spectra similar to those for the AP function. The optimal pH ranges for these two functions are also similar to each other (5.5-6.5). Six mutant HAP1 proteins containing single amino acid substitutions were assayed in parallel for comparison of their activities toward p-BQ-dC and the AP site. These mutants either concomitantly lost (N212A, D210N) or had reduced (D219A, E96A, and N212Q) or unchanged (H116N) p-BQ-dC and AP activities. This parallelism strongly supports the hypothesis that cleavage of p-BQ-dC requires the same catalytic active site as that proposed for the AP function. This dual activity toward two structurally unrelated substrates, an AP site and a bulky exocyclic adduct, has implications for substrate recognition. The AP site and p-BQ-dC cause different changes in the local conformation around the lesion as it is visualized by molecular modeling. PMID- 9398272 TI - Time-resolved fluorescence of O-acetylserine sulfhydrylase catalytic intermediates. AB - The reaction of the substrate O-acetyl-L-serine (OAS) with the pyridoxal 5' phosphate (PLP)-dependent enzyme O-acetylserine sulfhydrylase-A (OASS-A) proceeds via the transient formation of an external aldimine absorbing at 420 nm and a stable alpha-aminoacrylate intermediate absorbing at 330 and 465 nm. Stable external aldimine species are obtained by reaction of the enzyme with either the reaction product L-cysteine or the product analog L-serine. Static and time resolved fluorescence emission properties of the coenzyme in the above catalytic intermediates have been used to directly probe the active site conformation at different stages of the catalytic pathway. Upon excitation at either 420 or 330 nm, the external aldimines with L-cysteine and L-serine exhibit a structured emission centered at 490 nm with a shoulder at 530 nm. Fluorescence decays upon excitation at 420 nm are best fitted using two components with lifetimes of 1.1 and 3.8 ns, with the fractional intensity of the slow component being 0.92 with L cysteine and 0.75 with L-serine, respectively. The fast component, emitting at 530 nm, is attributed to a dipolar species formed in the excited state by proton dissociation, and the slow component, emitting at 490 nm, is attributed to a ketoenamine tautomer of the external aldimine. The slow component for external aldimine fluorescence decay is characterized by the same lifetime value as that of the internal aldimine with an increased fractional intensity, indicating that the distribution between the ketoenamine and the dipolar species is shifted toward the ketoenamine tautomer in the external aldimine, compared to the internal aldimine. Differences in equilibrium distribution of ketoenamine and enolimine tautomers can also account for differences in the emission properties of the external aldimines of L-cysteine and L-serine. The alpha-aminoacrylate species is characterized by a relatively weak emission. Upon excitation at 330 nm, the emission exhibits two bands centered at 420 and 540 nm, whereas upon excitation at 420 nm the emission bands are centered at 500 and 540 nm, and upon excitation at 465 nm, the main absorbance peak of the alpha-aminoacrylate species, the emission spectrum shows a band at 540 nm. The fluorescence decays, upon excitation at 330 nm, are best fitted using three components with lifetime values similar to those found for the internal aldimine, with the slow component predominating. Species-associated spectra, collected between 400 and 520 nm upon excitation at 350 nm, indicate the presence of a fast component overlapping the slow component on the blue side of the emission spectrum, as detected for the internal aldimine. When the excitation wavelength is 420 nm, there are only two components with the fast one predominating. A further increase in the fractional intensity of the fast component is observed upon excitation at 465 nm. The weak emission and the short lifetime of the emission excited at 465 nm indicate that this alpha-aminoacrylate tautomer interacts significantly with neighboring groups of the protein matrix and may be endowed with a higher mobility than the external aldimine. PMID- 9398273 TI - Allosteric interactions between DNA strands and monovalent cations in DNA quadruplex assembly: thermodynamic evidence for three linked association pathways. AB - The series of cooperative transitions that lead to [d(TG4)4.(K+)m] quadruplex assembly upon rapid addition of KCl to d(TG4) strands were studied. Quadruplex samples were dialyzed against KCl then Li-EDTA and found to retain between three and five strongly bound potassiums with affinities >10(6) M-2. Absorbance thermal denaturation (melt) and circular dichroism (CD) equilibrium binding data were obtained. The latter were analyzed using two classes of binding models to simulate the effects of the assumed intermolecular interactions on the binding curves (isotherms). The melt experiments yielded equilibrium dissociation constants (Kd) ranging from 10(-11) to 10(-12) M3 at the melting temperatures. Extrapolating these values to 23 degrees C predicts Kd values in the 10(-28) M3 range if the heat capacity (Cp) is not strongly dependent upon temperature changes over this range. Assuming Ka is equal to 1/Kd (from melting analyses), very large association free energies stabilize the quadruplex at 23 degrees C in 100 mM KCl (DeltaGa = -43 kcal mol-1). Plots of the differential melt curve peak half-widths, a measure of cooperativity, versus d(TG4) concentration showed that quadruplex dissociation is much more cooperative at 400 mM KCl than at 100 mM KCl. Forty-eight hour quadruplex assembly time courses were monitored by CD at 264 nm. Equilibrium quadruplex accumulation generally required over 10 h, and net reaction extents were in the 10-85% range. Hill plots of the data show that initial steps in the multistep pathway are positively cooperative, presumably due to strong strand-cation and strand-strand binding interactions in duplex and triplex assembly reactions, then negatively cooperative in quadruplex formation. Models were developed to rationalize the experimental observations in terms of consecutive cooperative allosteric transitions from cation-deficient relaxed (R) strand-aggregates to cation-containing tense (T) structures, driven by the allosteric effector K+. Quantitative mappings of positive and then negative cooperativity were obtained by fitting the results as a function of strand number incorporated during quadruplex assembly. Surprisingly, models for reactions involving incorporation of five and six strands fit the data better than models involving only four strands. The 5-step "induced fit" model fits the data as well as or better than 3- and 4-step models and better than all of the strand aggregation models that were devised and investigated. Net association free energies (summation operatori=1,n) ranged from -20 to -26 kcal mol-1, approximately half the magnitude of the apparent stabilities measured by absorbance melts. Likely explanations for this discrepancy involve hysteresis and errors due to inadequate equilibration in the melt experiments. Hysteresis is thought to be produced by irreversibility due to different predominant mechanisms in absorbance (dissociation) and CD (association) experiments. The kinetic block to quadruplex assembly can be unambiguously attributed to quadruplex formation and not intermediate steps in the assembly mechanism. On the basis of these results we propose that, in addition to the more conventional assembly mechanisms involving duplex dimerization and stepwise strand addition, quadruplex formation can also proceed by triplex-triplex disproportionation. Interaction statistics arguments that support the energetic feasibility of the disproportionation pathway are presented. The allosteric quadruplex assembly model provides a mechanism which could be used by the cell to simultaneously modulate DNA structure and activity within telomeres, transcriptional promoters, recombination prone chromatin, and other G-rich DNAs. As a result of this allosterism, cation and strand availability and strand-pairing capabilities could profoundly influence the functional capacity of a particular strand over a relatively narrow range of effector concentration changes. (ABSTRACT TRUNCATED) PMID- 9398274 TI - A leucine zipper motif in the ectodomain of Sendai virus fusion protein assembles in solution and in membranes and specifically binds biologically-active peptides and the virus. AB - We have detected a leucine zipper-like motif in the ectodomain of the Sendai virus fusion protein (aa 269-307) which is extremely conserved in the family of Sendai viruses. To find a possible role for this motif, we synthesized SV-269, a 39 amino acid peptide corresponding to this domain, and a mutant peptide, MuSV 269, with an amino acid pair interchanged their positions. The peptides were labeled with fluorescent probes at their N-terminal amino acid and functionally and structurally characterized. The data show that SV-269, but not MuSV-269, specifically binds Sendai virus. Expectedly, SV-269 is more active than the mutant MuSV-269 in inhibiting Sendai virus-mediated hemolysis. Fluorescence studies reveal that SV-269 assembles in aqueous solution, binds to zwitterionic PC and negatively-charged PS/PC vesicles, and assembles therein. Although MuSV 269 similarly binds to both types of vesicles, it only slightly assembles in solution and not at all in membranes. Moreover, SV-269, but not MuSV-269, coassembles with the biologically-active heptad repeats SV-150 and SV-473 (Rapaport et al. , 1995) in solution as revealed by fluorescence and circular dichroism (CD) spectroscopy, and with SV-150 within negatively-charged PS/PC and zwitterionic PC vesicles. Despite these differences, both SV-269 and MuSV-269 adopt similar secondary structures in 40% TFE and 1% SDS as revealed by CD spectroscopy, and disrupt the packing of the lipid bilayers to the same extent, as shown by the dissipation of diffusion potential. The role of this leucine zipper motif is discussed in terms of the assembly of the Sendai virus fusion protein in solution and within membranes. Since most of the heptadic leucines are also conserved in the corresponding domains of other paramyxoviruses such as rinderpest, measles, SV5, and parainfluenza, it may indicate a similar role of this domain in these viruses as well. PMID- 9398275 TI - FTIR spectroscopic studies of oligonucleotides that model a triple-helical domain in self-splicing group I introns. AB - Fourier Transform infrared (FTIR) spectroscopy was used to characterize the Mg2+ dependent association of a 23-mer mixed ribo-deoxyribonucleotide (23-mer RNA) and a 7-mer oligoribonucleotide (7-mer RNA) that models the triple-helical domain of a self-splicing group I intron [Sarkar et al. (1996) Biochemistry 35, 4678-4688]. To elucidate the effect of deoxyribose substitution in the entire backbone, as well as at specific positions, in the assembly of the triple-helical domain, parallel studies were carried out on the association of pure deoxyribonucleotides having base sequences corresponding to the oligoribonucleotides and also between 23-mer RNA and two 7-mer RNA variants. In the variants, either the ribose attached to G451 or the ribose attached to U453 was changed to deoxyribose. FTIR monitored thermal denaturation of the two 23-mer hairpins shows two distinct melting regions in 1 M NaCl, in case of the RNA hairpin but not for the 23-mer DNA. Triple-helix association between the two strands (7-mer and 23-mer) studied by FTIR show that only when both strands are RNA, association takes place with the formation of the P6 helix. Our results also show that the interactions between the two RNA strands involve some participation of the riboses, which could also involve the 2'-OH groups of the RNA backbone. The assembly of the triple-helical domain is not possible with a deoxyribose backbone and is completely perturbed even when only one ribose at either G451 or U453 position is substituted by deoxyribose. PMID- 9398276 TI - Detection and identification of transient enzyme intermediates using rapid mixing, pulsed-flow electrospray mass spectrometry. AB - Rapid chemical quench methods coupled with off-line detection have proven to be very useful in identifying enzyme reaction intermediates. However, a limitation to this approach involves enzyme intermediates which are too labile under the chemical quenching conditions to allow detection and characterization. In this report, we describe the development of a novel approach for the detection and characterization of enzyme intermediates on the subsecond time scale using a "pulsed flow" method which employs a direct interface between a rapid-mixing device and electrospray ionization mass spectrometry. The application of this technique with the enzyme 5-enolpyruvoyl-shikimate-3-phosphate (EPSP) synthase is demonstrated. This enzyme converts shikimate-3-phosphate (S3P) and phosphoenol pyruvate (PEP) to EPSP and inorganic phosphate. Previous rapid chemical quench studies have shown that this reaction proceeds through a tetrahedral intermediate [Anderson, K. S., et al. (1988) J. Am.Chem. Soc. 110, 6577-6579] formed transiently at the enzyme active site. We have shown that this tetrahedral intermediate can be directly detected on a subsecond time scale without chemical quenching by interfacing a rapid mixing apparatus directly with an on-line electrospray ionization ion trap mass spectrometer. Negative ion mass spectra collected by electrospray ionization indicate peaks for S3P (m/z 253), PEP (m/z 167), EPSP (m/z323), and the tetrahedral intermediate (m/z 421). Further confirmation was provided by performing the same experiment with [13C-1]-labeled PEP. These spectra confirmed the anticipated shift of 1 atomic mass unit for PEP (m/z 168), EPSP (m/z 324), and the tetrahedral intermediate (m/z 422) with no change in S3P (m/z 253). The collision-induced dissociation of the unlabeled tetrahedral intermediate peak (m/z421) produced a daughter ion at m/z 323, which is most likely EPSP resulting from the loss of phosphate and is consistent with previous studies which have examined the chemical breakdown of the tetrahedral intermediate in solution [Anderson, K. S., et al. (1990) J. Biol. Chem. 265, 5567 6672]. This technique is under development and should be a useful method to study the transient formation of enzyme intermediates. PMID- 9398277 TI - Kinetic and spectroscopic investigations of wild-type and mutant forms of apple 1 aminocyclopropane-1-carboxylate synthase. AB - Two catalytically inactive mutant forms of 1-aminocyclopropane-1-carboxylate (ACC) synthase, Y85A and K273A, were mixed in low concentrations of guanidine hydrochloride (GdnHCl). About 15% of the wild-type activity was recovered (theoretical 25% for a binomial distribution), proving that the functional unit of the enzyme is a dimer, or theoretically, a higher order oligomer. The enzyme catalyzes the conversion of S-adenosyl-L-methionine (SAM) to ACC. The value of kcat/KM is 1.2 x 10(6) M-1 s-1 at pH 8.3. Viscosity variation experiments with glycerol and sucrose as viscosogenic reagents showed that this reaction is nearly 100% diffusion controlled. The sensitivity to viscosity for the corresponding reaction of the less reactive Y233F mutant is much reduced, thus the latter reaction serves as a control for that of the wild-type enzyme. The kcat/KM vs pH profile for wild-type enzyme exhibits pKa values of 7.5 and 8.9. The former is assigned to the pKa of the alpha-amino group of SAM, while the latter corresponds to the independently determined spectrophotometric pKa of the internal aldimine. The kcat vs pH profile exhibits similar pKas, which means that the above pKa values are not perturbed in the Michaelis complex. The phenolic hydroxyl group of Tyr233 forms a hydrogen bond to the 3'-O- of PLP. The spectral and kinetic pKa (kcat/KM) values of the Y233F mutant are not identical (spectral 10.2, kinetic 8.7). A model that accounts quantitatively for these data posits two parallel pathways to the external aldimine for this mutant, the minor one has the alpha amino group free base form of SAM reacting with the protonated imine form of the enzyme with kcat/KM approximately 6.0 x 10(3) M-1 s-1, while the major pathway involves reaction of the aldehyde form of PLP with SAM with kcat/KM approximately 7.0 x 10(5) M-1 s-1. The spectral pKa is defined only by the less reactive species. PMID- 9398278 TI - Evidence that galactanase A from Pseudomonas fluorescens subspecies cellulosa is a retaining family 53 glycosyl hydrolase in which E161 and E270 are the catalytic residues. AB - A genomic library of Pseudomonas fluorescens subsp. cellulosa DNA was screened for galactanase-positive recombinants. The nine galactanase positive phage isolated contained the same galactanase gene designated galA. The deduced primary structure of the enzyme (galactanase A; GalA) encoded by galA had a Mr of 42 130 and exhibited significant sequence identity with a galactanase from Aspergillus aculeatus, placing GalA in glycosyl hydrolase family 53. The enzyme displayed properties typical of an endo-beta1, 4-galactanase and exhibited no activity against the other plant structural polysaccharides evaluated. Analysis of the stereochemical course of 2,4-dinitrophenyl-beta-galactobioside (2,4-DNPG2) hydrolysis by GalA indicated that the galactanase catalyzes the hydrolysis of glycosidic bonds by a double displacement general acid-base mechanism. Hydrophobic cluster analysis (HCA) suggested that family 53 enzymes are related to the GH-A clan of glycosyl hydrolases, which have an (alpha/beta)8 barrel structure. HCA also predicted that E161 and E270 were the acid-base and nucleophilic residues, respectively. Mutants of GalA in which E161 and E270 had been replaced with alanine residues were essentially inactive against galactan. Against 2,4-DNPG2, E161A exhibited a much lower Km and kcat than native GalA, while E270A was inactive against the substrate. Analysis of the pre-steady-state kinetics of 2,4-DNPG2 hydrolysis by E161A showed that there was an initial rapid release of 2,4-dinitrophenol (2,4-DNP), which then decayed to a slow steady-state rate of product formation. No pre-steady-state burst of 2,4-DNP release was observed with the wild-type enzyme. These data are consistent with the HCA prediction that E161 and E270 are the acid-base and nucleophilic catalytic residues of GalA, respectively. PMID- 9398279 TI - Evidence for interaction between transmembrane segments in assembly of Kv1.3. AB - Previously, we showed that the N-terminal recognition domain (T1) of Kv1.3 was not required for assembly of functional channels [Tu et al. (1996) J. Biol. Chem. 271, 18904-18911]. Moreover, specific Kv1.3 peptide fragments including regions of the central core are able to inhibit expression of current produced from a channel lacking the T1 domain, Kv1.3(T1-). To elucidate the mechanism whereby Kv1.3 peptide fragments suppress Kv1.3(T1-) current, we have studied the ability of peptide fragments containing the transmembrane segments S1, S1-S2, or S1-S2-S3 to physically associate with the Kv1.3(T1-) polypeptide subunit in vitro in microsomal membranes. Using c-myc (9E10) epitope-labeled peptide fragments and anti-myc antibody as well as antisera to the Kv1.3 C-terminus, we now demonstrate specific association of these peptide fragments with Kv1.3(T1-). Association of peptide fragments with Kv1.3(T1-) was correlated with integration of both proteins into the membrane. Furthermore, the relative strength and kinetics of this association directly correlated with the ability of fragments to suppress Kv1.3(T1-) current. The rate-limiting step in the sequential synthesis, integration, and formation of a complex was the association of integrated polypeptides within the plane of the lipid bilayer. These results strongly suggest that the physical association of transmembrane segments provides the basis for suppression of K+ channel function by K+ channel peptide fragments in vivo. Moreover, the S1-S2-S3 peptide fragment potently suppressed full-length Kv1.3, thus implicating a role for the S1-S2-S3 region of Kv1.3 in the assembly of the Kv1.3 channel. We refer to these putative association sites as IMA (intramembrane association) sites. PMID- 9398280 TI - Affinity cleavage at the metal-binding site of phosphoenolpyruvate carboxykinase. AB - Chicken liver phosphoenolpyruvate carboxykinase (PEPCK) was rapidly inactivated by micromolar concentrations of ferrous sulfate in the presence of ascorbate at pH 7.4. Omitting ascorbate or replacing the Fe2+ with Mn2+ or Mg2+ gives no inactivation. Mn2+, Mg2+, or Co2+ at 100-fold molar excess over Fe2+ offered complete protection from Fe2+/ascorbate-induced inactivation. The substrates PEP and GTP, but not OAA, GDP, or CO2, offered full protection from inactivation. The addition of 5 mM EDTA stopped further inactivation of the enzyme. Thermodynamic studies indicate that the inactive enzyme no longer binds Mn2+ but still had high affinity for GTP indicating that the inactivation process was specific for the metal site. A decrease in cysteine content was observed over time following PEPCK treatment with Fe2+ and ascorbate. The apparent first-order rate constant for free sulfhydryl loss (0.085 +/- 0.005 min-1) is similar to the apparent first order rate constant for inactivation (0.067 +/- 0.005 min-1). Amino acid composition analysis revealed that cysteic acid was generated upon Fe2+/ascorbate addition to PEPCK. Native chicken liver PEPCK has an Mr of 67 kDa. SDS-PAGE of the inactivated enzyme showed the presence of two new bands at 31.7 and 35.3 kDa indicating that PEPCK was specifically cleaved at a single site. The rate of cleavage was slower than the rate of inactivation and fully inactivated enzyme was only 50% cleaved. The Fe2+/ascorbate-catalyzed inactivation was not solely due to protein cleavage. The protein fragments generated by cleavage were separated by C4 reverse phase HPLC. The cleavage exposed a new N-terminus which was identified to be the 35.3 kDa C-terminal half of PEPCK. Sequencing of the fragments indicated that the site of cleavage was between Asp296 and Ile297. These results indicate that Asp296 is involved in metal chelation. This agrees with previous studies [Hlavaty, J. J., & Nowak, T. (1997) Biochemistry 36, 3389 3403] that suggested that Asp295 and Asp296 are involved in metal binding. PMID- 9398281 TI - One- and two-dimensional ESEEM spectroscopy of flavoproteins. AB - One- and two-dimensional (1D and 2D) electron spin echo envelope modulation (ESEEM) spectroscopy was applied to study the flavin cofactors in the neutral semiquinone states of flavodoxin and ferredoxin-NADP+ reductase (FNR) from the cyanobacterium Anabaena PCC 7119, and the anionic semiquinone state of cholesterol oxidase from Brevibacterium sterolicum. High-resolution crystal structures are available for all these proteins. Three- and 4-pulse ESEEM and hyperfine sublevel correlation spectroscopy (HYSCORE) techniques at X-band were used. HYSCORE spectra showed correlations between transitions caused by interaction of the isoalloxazine unpaired electronic spin present in the semiquinone state with several nitrogen and hydrogen nuclei. Measurements of isotopic labeled samples ([15N]FMN flavodoxin and [2H]flavodoxin) allowed the assignment of all the detected transitions to nuclei belonging to the FMN cofactor group. Interactions of nitrogens in positions 1 and 3 of the isoalloxazine ring were determined to have isotropic hyperfine coupling constants in the 1-2 and 0.5-1 MHz ranges for all the different flavoprotein semiquinones studied. Information about the quadrupolar term of these nuclei was also obtained. An intense correlation in the negative quadrant was detected. It has been associated to the strongly interacting N(10) nucleus. The complete hyperfine term parameters (including the sign) were obtained from detailed analysis of this signal, being the quadrupolar parameter, K, also estimated. Another correlation in the HYSCORE spectra, corresponding to hydrogen bound to the N(5) position in neutral flavin semiquinones, was detected. Its interaction parameters were also determined. This study demonstrates that ESEEM spectroscopy, and in particular the HYSCORE technique, are of particular utility for detecting and assigning nuclear transition frequencies in flavoprotein semiquinones. Moreover, the results reported here are complementary to ENDOR studies, and both techniques together provide an important tool for obtaining information about spin distribution in the flavin ring of flavoproteins in the semiquinone state. PMID- 9398282 TI - Special folding pathway to tetramer only through the micelle state of the corticotropin-releasing factor. AB - The ovine corticotropin-releasing factor (CRF), a peptide hormone of 41 residues stimulating the secretion of adrenocorticotropic hormone, was thermodynamically investigated. By means of size exclusion chromatography and/or ultrafiltration, the CRF solution could be separated into random coil monomers and highly alpha helical tetramers, which seem to have amphipathic helix bundle structure. Circular dichroism measurements along with diluting or concentrating the CRF solution revealed that there exists the micelle state above the concentration of 0.1 mM, which would be the critical micelle concentration (cmc). The micelle state was also proved by binding ability for 8-anilino-1-naphthalenesulfonate and endothermic change by dilution across the cmc. The tetramer showed the cooperative thermal transition at about 55 degrees C in the buffer solution (pH 7.5), so that it would have native-protein-like folding. On the other hand, the micelle undergoes gradual change to dissociated state by heating, regardless of the similar alpha-helicity to the tetramer. Above the cmc the equilibrium between the tetramer and the micelle takes place as well as that between the monomer and the micelle. Whereas, the direct conversion between the tetramer and the monomer scarcely occurred below the cmc. The titration experiment with 2,2,2 trifluoroethanol (TFE) revealed that the cmc decreases with increasing the concentration of TFE. This tendency is the same as that of general surfactants. Most of experimental results can be well explained by this three-phase model involving the monomer, the tetramer, and the micelle. The lack of the equilibrium between the monomer and the tetramer indicates that the folding pathway of the tetramer is the transformation only through the micelle state and not from the monomer. This pathway resembles the collapse model among the phenomenological models for thermodynamic protein folding. By the mathematical consideration for the dissociation of micelle, we have demonstrated that the expected content of undegradable k-mer is 2/(k + 1), which agreed well with the observed tetramer content of CRF (40%). PMID- 9398283 TI - Bacillus thuringiensis cytolytic toxin associates specifically with its synthetic helices A and C in the membrane bound state. Implications for the assembly of oligomeric transmembrane pores. AB - The CytA toxin exerts its activity by the formation of pores within target cell membranes. However, the exact mechanism of pore formation and the structural elements that are involved in the toxic activity are yet to be determined. Recently, the structure of the highly similar CytB toxin was solved (Li et al., 1996), and a beta-barrel was suggested as a possible structure of the pores. Due to the similarity between the toxins, the existence and positioning of alpha helices and beta-sheets in CytA were predicted from the alignment of the sequences. Here peptides corresponding to beta5, beta6, and beta7 strands, to a conserved nonhelical region of the CytA toxin (P149-170), to helices B and D, and to an analogue of helix A were synthesized, fluorescently labeled, and characterized. We found that, unlike helices A and C (Gazit and Shai, 1993), neither the beta-strand peptides nor helix B could interact with lipid membranes, whereas P149-170 and helix D bind the membrane weakly. Membrane permeation experiments suggested that CytA toxin exerts its activity by aggregation of several monomers. To learn about the structural elements that may mediate CytA oligomerization, the ability of the synthetic peptides to interact with membrane bound CytA was studied. Helices A and C, but not the beta-strands, helix D, or a control peptide, caused a large increase in the fluorescence of membrane-bound fluorescein-labeled CytA, whereas helix B had only a slight effect. Moreover, the addition of rearranged helix A, a peptide with the same composition as helix A, but with only two pairs of amino acids rearranged, did not affect the fluorescence. The addition of unlabeled CytA also caused an increase in the fluorescence intensity, further demonstrating the interaction between CytA monomers within the membrane. Taken together, our results provide further support for the suggestion that the CytA toxin self-assembles within membrane and that helices A and C are major structural elements involved in the membrane interaction and intermolecular assembly of the toxin. PMID- 9398284 TI - Purealin blocks the sliding movement of sea urchin flagellar axonemes by selective inhibition of half the ATPase activity of axonemal dyneins. AB - Ciliary and flagellar movements are explained by active sliding between the outer doublet microtubules of an axoneme via their inner and outer dynein arms. Purealin, a novel bioactive principle of a sea sponge Psammaplysilla purea, blocked the motility of Triton-demembranated sea urchin sperm flagella within 5 min at concentrations above 20 microM. In a similar concentration range, purealin blocked the sliding movement of the flagellar axonemes in vitro within a few minutes judging from the turbidity measurements. The ATPase activity of axonemes was partially inhibited by purealin in a concentration-dependent manner. The maximum inhibition reached approximately 50% at concentrations above 20 microM, indicating that half the axonemal ATPase activity is sensitive to purealin. Similar results were observed on the ATPase activity of outer-arm-depleted axonemes and that of a mixture of 21S dynein and salt-extracted axonemes. On the other hand, ATPase activity of isolated 21S dynein was not inhibited by purealin. The inhibitory action of purealin on the axonemal ATPases was reversed by dilution of purealin. The effect of purealin on the double-reciprocal plot of the ATPase activity as a function of ATP concentrations showed that the inhibition was not a competitive type. In accord with this finding, purealin did not affect the vanadate-mediated UV photocleavage of axonemal dyneins. These results suggest that purealin binds reversibly to a site other than the catalytic ATP-binding site and inhibits half the ATPase activity of axonemes. Taken together, our results suggest that purealin-sensitive ATPase activity of the dynein arms plays an essential role in generating the sliding movement of flagellar axonemes. PMID- 9398285 TI - Participation of the N-terminal region of Cepsilon3 in the binding of human IgE to its high-affinity receptor FcepsilonRI. AB - The binding of immunoglobulin E (IgE) to its high-affinity receptor (FcepsilonRI) expressed on mast cells and basophils is central to the development of an allergic reaction. Previous studies have implicated the third constant domain of IgE-Fc (Cepsilon3) as the site of the interaction with FcepsilonRI. We have prepared a series of site-directed mutants of human IgE-Fc, particularly focusing on the N-terminal "linker" region and AB loop of Cepsilon3. The kinetics of binding IgE and its Fc fragments to the immobilized receptor were determined by surface plasmon resonance (SPR), and two phases of binding were observed. We identified one mutation in the N-terminal linker region, R334S, that has a dramatic effect on binding. R334S lowers the affinity of IgE-Fc for FcepsilonRI by 120-fold, principally through an increase in the dissociation rate of the slower phase of the interaction. This mutation has a similar effect in Fcepsilon3 4, a truncated form of IgE-Fc which lacks the Cepsilon2 domain pair, and thus it does not exert its effect through altering the quaternary structure of IgE-Fc, firmly implicating Arg334 as a contact residue in the complex. However R334S has no effect on the binding of FcepsilonRII (CD23), the low-affinity receptor for IgE, demonstrating the structural integrity of the mutated IgE-Fc. Circular dichroism spectroscopy and thermal stability studies further indicate that the R334S mutation does not disorder or destabilize the structure of IgE-Fc or Fcepsilon3-4. These results demonstrate the importance of the N-terminal linker region of Cepsilon3 in the interaction of IgE with FcepsilonRI. PMID- 9398286 TI - Identification of contact residues in the IgE binding site of human FcepsilonRIalpha. AB - The high-affinity receptor for immunoglobulin E (IgE), FcepsilonRI, is an alphabetagamma2 tetramer found on mast cells, basophils, and several other types of immune effector cells. The interaction of IgE with the alpha-subunit of FcepsilonRI is central to the pathogenesis of allergy. Detailed knowledge of the mode of interaction of FcepsilonRI with IgE may facilitate the development of inhibitors for general use in the treatment of allergic disease. To this end we have performed site-directed mutagenesis on a soluble form of the FcepsilonRI alpha-chain (sFcepsilonRIalpha). The effects of four mutations in the second immunoglobulin-like domain of sFcepsilonRIalpha upon the kinetics of binding to IgE and fragments of IgE have been analyzed using surface plasmon resonance. As described in the preceding paper of this issue [Henry, A. J., et al. (1997) Biochemistry 36, 15568-15578], biphasic binding kinetics was observed. Two of the mutations had significant effects on binding: K117D reduced the affinity of sFcepsilonRIalpha for IgE by a factor of 30, while D159K increased the affinity for IgE by a factor of 7, both principally through changes in the rates of dissociation of the slower phase of the interaction. Circular dichroism spectra of sFcepsilonRIalpha incorporating either of these mutations were indistinguishable from those of wild-type sFcepsilonRIalpha, demonstrating that the native conformation had not been disrupted. Our results, together with those from site-directed mutagenesis on fragments of IgE presented in the accompanying paper, define the contact surfaces in the IgE:sFcepsilonRIalpha complex. PMID- 9398287 TI - Ionic properties of membrane association by vitamin K-dependent proteins: the case for univalency. AB - Ionic properties of membrane interaction by prothrombin, protein Z, and other vitamin K-dependent proteins were studied to determine the relevance of a monovalent membrane contact mechanism between one phospholipid headgroup and a calcium-lined pore in the protein [McDonald, J. F., Shah, A. M., Schwalbe, R. A., Kisiel, W., Dahlback, B., and Nelsestuen, G. L. (1997) Biochemistry 36, 5120 5127]. For comparison, multivalent ionic interaction was illustrated by peptides of +3 to +5 net charge and by blood clotting factor V. As expected, the peptides were easily dissociated by salt and gave nominal charge-charge interactions (zazb values) of -13 to -17. Factor V showed much higher binding affinity despite nominal zazb values of about 9. Membrane-bound prothrombin and protein Z showed very low sensitivity to salt as long as calcium was at saturating levels (zazb values of approximately -1.3 to -1.4), appropriate for univalent ionic attraction. Prothrombin contains +3 charge groups (Lys-2, Lys-11, Arg-10) that are absent from the GLA domain (residues 1-35) of protein Z, while protein Z contains -4 charge groups (Gla-11, Asp-34, Asp-35) that are absent in prothrombin. Thus, similar zazb relationships indicated little role for these surface charges in direct membrane contact. Calcium-saturated protein Z bound to phosphatidylcholine (PC) in a manner which indicated the addition of one calcium ion, bringing the total calcium stoichiometry in the protein-membrane complex to at least 8. Protein Z bound to phosphatidic acid (PA) in a manner suggesting the need for a fully ionized phosphate headgroup, a property expected by ion pairing in an isolated environment. Electrostatic calculations showed that the proposed protein site for phosphate interaction was electropositive. The cluster of hydrophobic amino acids (Phe-5, Leu-6, and Val-9) on the surface of prothrombin was electronegative, suggesting a role in the electrostatic architecture of the GLA domain. Overall, membrane binding by vitamin K-dependent proteins appeared consistent with the formation of an ion pair in an isolated environment. PMID- 9398288 TI - pH dependence of antibody/lysozyme complexation. AB - Association between proteins often depends on the pH and ionic strength conditions of the medium in which it takes place. This is especially true in complexation involving titratable residues at the complex interface. Continuum electrostatics methods were used to calculate the pH-dependent energetics of association of hen egg lysozyme with two closely related monoclonal antibodies raised against it and the association of these antibodies against an avian species variant. A detailed analysis of the energetic contributions reveals that even though the hallmark of association in the two complexes is the presence of conserved charged-residue interactions, the environment of these interactions significantly influences the titration behavior and concomitantly the energetics. The contributing factors include minor structural rearrangements, buried interfacial area, dielectric environment of the key titratable residues, and geometry of the residue dispositions. Modeled structures of several mutant complexes were also studied so as to further delineate the contribution of individual factors to the titration behavior. PMID- 9398289 TI - Regulation of protein kinase C betaII by its C2 domain. AB - The C2 domain serves as a membrane-targeting module in a diverse group of proteins that includes the conventional protein kinase Cs. This work examines the mechanism by which the C2 domain targets protein kinase C to membranes. Molecular modeling identified two highly-charged surfaces on the C2 domain of protein kinase C betaIotaIota: the Ca2+ binding site which contains five aspartates and a basic face positioned behind the Ca2+ site that contains seven lysine residues. Both surfaces were mutated to assess their role in Ca2+-dependent membrane binding. Surprisingly, removal of four positive charges on the basic face had no effect on protein kinase C's lipid or Ca2+ sensitivity, revealing that the basic face does not provide determinants involved in lipid binding, nor is it positioned close enough to the membrane to enhance nonspecific recruitment by its electropositive face. In contrast, replacement of two negative charges with two positive charges in the Ca2+ binding site decreased protein kinase C's affinity both for Ca2+ and for anionic lipids by several orders of magnitude. The dramatic reduction in electronegative potential resulting from this mutation did not increase protein kinase C's affinity for acidic membranes in the absence of Ca2+, revealing that simple charge neutralization does not account for how Ca2+ increases protein kinase C's affinity for anionic membranes. Our data suggest that (1) the membrane interaction surface of the C2 domain is localized to the Ca2+-binding site, with the positive face positioned away from the membrane, and (2) the Ca2+ site does not serve as a simple electrostatic switch. PMID- 9398290 TI - Mapping the suramin-binding sites of human neutrophil elastase: investigation by fluorescence resonance energy transfer and molecular modeling. AB - Neutrophil elastase (NE), a mediator of inflammation, binds with high affinity numerous anionic molecules including suramin, a polysulfated naphthylurea, which inhibits it with a Ki of 0.2 microM and a 4:1 suramin:NE stoichiometry and thus constitutes a potential therapeutic agent. In an attempt to locate the suramin molecules on NE, we investigated the NE-suramin interaction using steady-state and time-resolved fluorescence spectroscopy. The time-resolved intensity decay of NE, a protein with three Trp residues, in positions 27, 141, and 237 (chymotrypsin numbering system) was best described by a three-exponential function with lifetimes ranging from 0.22 to 2.28 ns. Comparison of the accessibility of the three lifetime classes to the fluorescence quenchers acrylamide and iodide with the computed solvent accessibility of the three Trp residues in the crystal structure of NE indicates that the main, if not the sole, contribution to the 2.28 ns lifetime class is brought about by the fully buried Trp 141 residue. The addition of suramin to NE induces a sharp decrease in NE fluorescence and a corresponding increase in suramin fluorescence due to an efficient fluorescence resonance energy transfer (FRET) between the Trp residues of NE, acting as donors, and the naphthalene rings of suramin, behaving as acceptors. From the fate of the longest lifetime class in the presence of variable suramin concentrations, we deduce that two suramins are bound at less than 17 A from Trp 141, whereas the two others are located at least 29 A from Trp 141. Moreover, neither the binding of suramin to NE nor the FRET process was modified when NE was complexed with a peptide chloromethylketone inhibitor, suggesting that suramin does not directly interfere with the substrate binding site of NE. These data were used as constraints to model the NE-suramin complex. PMID- 9398291 TI - Ligand-induced conformational changes in lactose repressor: a fluorescence study of single tryptophan mutants. AB - A key element in the ability of lac repressor protein to control transcription reversibly is the capacity to assume different conformations in response to ligand binding. To investigate regions of the protein involved in these conformational changes, mutant repressor proteins containing single tryptophans were created by mutating each of the two native tryptophan residues to tyrosine and changing the residue of interest to tryptophan. Tryptophans substituted in the following locations were highly accessible to quenchers with no changes in fluorescence or quenching properties in the presence of ligands: in the N terminal helix-turn-helix for Y7, at the junction between the N-terminus and N subdomain for L62, in the N-subdomain of the monomer-monomer interface for residue E100 or Q117, or at the C-terminal region for K325. Tryptophan at position F226 in the C-subdomain subunit interface was only moderately exposed to quenchers and unresponsive to ligands. In contrast, the fluorescence and quenching properties of single tryptophans placed in the central region of the protein were affected by ligands. Inducer binding altered the accessibility to quencher for tryptophan at H74 or F293, but no changes were detected upon binding operator. Exposure of tryptophan at the position occupied by Y273 was affected by both inducer and operator, indicating alterations in this region by both ligands. These results suggest that, in the areas of the lac repressor probed by these substitutions, the inducer-bound form differs from the conformation of the unliganded form. PMID- 9398292 TI - Identification of catalytically important residues in yeast transketolase. AB - The possible roles of four histidine residues in the active site of yeast transketolase were examined by site-directed mutagenesis. Replacement of the invariant His69 with alanine yielded a mutant enzyme with 1.5% of the specific activity of the wild-type enzyme and with an increased KM for the donor. This residue is located at the bottom of the substrate cleft close to the C1 hydroxyl group of the donor substrate, and the side chain of His69 might be required for recognition of this hydroxyl group and possibly for maintenance of the proper orientation of the reaction intermediate, (alpha, beta-dihydroxyethyl)thiamin diphosphate. Amino acid replacements of His481 by alanine, serine, and glutamine resulted in mutant enzymes with significantly increased KM values for the donor substrate and specific activities of 4.4%, 1.9%, and 5.5% of the wild-type enzyme. The kinetic data suggest that this residue, although close to the C2 carbonyl oxygen of the substrate, is not absolutely required for stabilization of the negative charge that develops at this oxygen in the transition state. This points toward the 4'-NH2 group of the pyrimidine ring of thiamin diphosphate as the major source of charge stabilization. Mutations at positions His30 and His263 result in mutant enzymes severely impaired in catalytic activity (1.5% and less of the activity of wild-type transketolase). The KM value for the donor substrate was increased for the His30Ala mutant but remained unchanged in the His263Ala enzyme. The side chains of both residues interact with the C3 hydroxyl group of the donor substrate, and the results indicate that the two residues act in concert during proton abstraction of the C3 hydroxyl proton during catalysis. PMID- 9398293 TI - Acyl-adenylate motif of the acyl-adenylate/thioester-forming enzyme superfamily: a site-directed mutagenesis study with the Pseudomonas sp. strain CBS3 4 chlorobenzoate:coenzyme A ligase. AB - 4-Chlorobenzoate:coenzyme A (4-CBA:CoA) ligase catalyzes 4-chlorobenzoyl-coenzyme A formation in a two-step reaction consisting of the adenylation of 4 chlorobenzoate with adenosine 5'-triphosphate followed by acyl transfer from the 4-chlorobenzoyl adenosine 5'-monophosphate diester intermediate to coenzyme A. In this study, two core motifs present in the Pseudomonas sp. strain CBS3 4-CBA:CoA ligase (motif I, 161T-S-G-T-T-G-L-P-K-G170, and motif II, 302Y-G-T-T-E306) and conserved among the sequences representing the acyl-adenylate/thioester-forming enzyme family (to which the ligase belongs) were tested for their possible role in substrate binding and/or catalysis. The site-directed mutants G163I, G166I, P168A, K169M, and E306Q were prepared and then subjected to steady-state and transient kinetic studies. The results, which indicate reduced catalysis of the adenylation of 4-chlorobenzoate in the mutant enzymes, are interpreted within the context of the three-dimensional structure of the acyl-adenylate/thioester forming enzyme family member, firefly luciferase. PMID- 9398294 TI - Structural and biochemical characterization of the GTPgammaS-, GDP.Pi-, and GDP bound forms of a GTPase-deficient Gly42 --> Val mutant of Gialpha1. AB - The Gly42 --> Val mutant of Gialpha1 was characterized structurally and biochemically to elucidate two important features of Gialpha1-catalyzed GTP hydrolysis. The crystal structure of the GTPgammaS-bound G42VGialpha1 protein demonstrates that the steric bulk of Val42 pushes the Gln204 residue into a catalytically incompetent conformation, providing a rationale for the diminished GTPase activity of this mutant. The same phenomenon may also account for the diminished GTPase activity of the homologous transforming Gly42 --> Val mutation in p21(ras). Similarly, the steric bulk of the unique Ser42 residue in Gzalpha may account for the comparatively slower rate of GTP hydrolysis by this Galpha subunit. The G42VGialpha1 subunit was also characterized structurally in its GDP.Pi- and GDP-bound states, providing a unique opportunity to view three "snapshots" of GTP hydrolysis. Hydrolysis of GTP to a transient GDP.Pi-bound intermediate is associated with substantial conformational changes in the switch II segment of the protein. Eventual release of Pi results in further removal of switch I from the active site and a highly mobile switch II segment. Despite their disparate biochemical properties, the structural similarity of G42VGialpha1 to the G203AGialpha1 mutant in the GDP.Pi-bound form suggests that both mutations stabilize a conformation of the GDP. Pi-bound protein that occurs only transiently in the wild-type protein. The structures of the GDP-bound forms of the wild-type and mutant proteins are similar. PMID- 9398295 TI - Role of the extracellular loops of the thyrotropin-releasing hormone receptor: evidence for an initial interaction with thyrotropin-releasing hormone. AB - Thyrotropin-releasing hormone (TRH), like most small ligands, appears to bind within the seven transmembrane-spanning helices (TMs) of its G protein-coupled receptor (TRH-R). A role for the extracellular loops (ECLs) of TRH-R has not been established. We substituted residues in the ECLs of TRH-R and show that Tyr-181 is important for high-affinity binding because its substitution leads to a 3700 fold lowering of the estimated affinity compared to wild-type TRH-R. Using TRH analogues, we provide evidence that there is a specific interaction between Tyr 181 in ECL-2 and the pyroGlu moiety of TRH. It was previously suggested that the pyroGlu of TRH may interact with Asn-110 in TM-3 and with Asn-289 in ECL-3; N110A and N289A TRH-Rs exhibit similar apparent affinities that are only 20-30-fold lower than wild-type TRH-R. To better understand these findings, we analyzed a computer-generated model which predicts that the ECLs form an entry channel into the TRH-R TM bundle, that Tyr-181 projects into this channel and that the pyroGlu of TRH cannot simultaneously interact with residues in the TMs and ECLs. Kinetic analysis showed that the association rate of [Ntau-methyl-His]TRH with N289A TRH R is slower than with wild-type TRH-R and largely accounts for the lower apparent affinity; the association rate with N110A TRH-R is similar to that of wild-type TRH-R. These data are consistent with the idea that there are initial interactions between TRH and the residues of a putative entry channel of TRH-R. We suggest that a role of the ECLs in all G protein-coupled receptors for small ligands may be to initially contact the ligand and allow entry into a TM binding pocket. PMID- 9398296 TI - Role of gamma-carboxyglutamic acid in the calcium-induced structural transition of conantokin G, a conotoxin from the marine snail Conus geographus. AB - Conantokin G is a gamma-carboxyglutamic acid- (Gla-) containing conotoxin isolated from the venom of the marine cone snail Conus geographus. This 17 residue polypeptide, which contains five gamma-carboxyglutamic acid residues, is a N-methyl-d-aspartate- (NMDA-) type glutamate receptor antagonist. To investigate the role of gamma-carboxyglutamic acid in the calcium-induced structural transition of conantokin G, we determined the three-dimensional structure of the conantokin G/Ca2+ complex by two-dimensional 1H NMR spectroscopy and compared it to the high-resolution structure of conantokin G in the absence of metal ions [Rigby et al. (1997) Biochemistry 36, 6906]. Complete resonance assignments were made by two dimensional 1H NMR spectroscopy at pH 5.6 in the presence of saturating amounts of Ca2+. Distance geometry and simulated annealing methods were used to derive 23 convergent structures from a set of 302 interproton distance restraints and two torsion angle measurements. A high resolution structure, with the backbone root mean square deviation to the geometric average of the 23 structures of 0.6 +/- 0.1 A, contains a linear alpha helix from Gla 3 to Lys 15. Gla residues 3, 7, 10, and 14 are aligned in a linear array on one face of the helix. A genetic algorithm was applied to determine the calcium positions in conantokin G, and the conantokin G/Ca2+ complex refined by molecular simulation. Upon binding of Ca2+ to gamma-carboxyglutamic acid, conantokin G undergoes a conformational transition from a distorted curvilinear 310 helix to a linear alpha-helix. Occupancy of the metal binding sites, defined by gamma-carboxyglutamic acids, results in formation of a calcium-carboxylate network that linearizes the helix and exposes the hydrophobic amino acids on the opposite face of the helix. PMID- 9398297 TI - Folding dynamics of the src SH3 domain. AB - The thermodynamics and kinetics of folding of the chicken src SH3 domain were characterized using equilibrium and stopped-flow fluorescence, circular dichroism (CD), and nuclear magnetic resonance (NMR) hydrogen exchange experiments. As found for other SH3 domains, guanidinium chloride (GdmCl) denaturation melts followed by both fluorescence and circular dichroism were nearly superimposable, indicating the concerted formation of secondary and tertiary structure. Kinetic studies confirmed the two-state character of the folding reaction. Except for a very slow refolding phase due to proline isomerization, both folding and unfolding traces fit well to single exponentials over a wide range of GdmCl concentrations, and no burst phase in amplitude was observed during the dead time of the stopped-flow instrument. The entropy, enthalpy, and heat capacity changes upon unfolding were determined by global fitting of temperature melts at varying GdmCl concentrations (0.4-3.7 M). Estimates of the free energy of unfolding, DeltaGUH2O, from guanidine denaturation, thermal denaturation, and kinetic experiments were in good agreement. To complement these data on the global characteristics of src SH3 folding, individual hydrogen-deuterium (HD) exchange rates were measured for approximately half of the backbone amides in 0 and 0.7 M GdmCl. The calculated free energies of the opening reaction leading to exchange (DeltaGHD) indicated that unfolding is highly cooperative--slowly exchanging protons were distributed throughout the core of the protein. The slowly exchanging protons exhibited DeltaGHD values higher than the global DeltaGUH2O by approximately 1 kcal/mol, suggesting that the denatured state might be somewhat compact under native conditions. Comparison of the src SH3 with homologous SH3 domains as well as with other small well-characterized beta-sheet proteins provides insights into the determinants of folding kinetics and protein stability. PMID- 9398298 TI - Three-dimensional solution structure of alpha-conotoxin MII, an alpha3beta2 neuronal nicotinic acetylcholine receptor-targeted ligand. AB - alpha-Conotoxin MII, isolated from Conus magus, is a potent peptidic toxin which specifically targets the mammalian neuronal nicotinic acetylcholine receptor, alpha3beta2 subtype. The three-dimensional structure of alpha-conotoxin MII in aqueous solution has been determined by two-dimensional 1H NMR spectroscopy. NOE derived distances, refined by an iterative relaxation matrix approach, as well as dihedral and chirality restraints were used in high-temperature biphasic simulated annealing calculations. Fourteen minimum energy structures out of 50 subjected to the SA simulations were chosen for evaluation; these 14 structures have a final RMS deviation of 0.76 +/- 0.31 and 1.35 +/- 0.34 A for the backbone and heavy atoms, respectively. The overall structure is unusually well-defined due to a large helical component around the two disulfide bridges. The principal backbone folding motif may be common to a subclass of alpha-conotoxins. There are two distinct surfaces on the molecule almost at right angles to one another. One entirely consists of the hydrophobic residues Gly1, Cys2, Cys3, Leu15, and Cys16. The second comprises the hydrophilic residues Glu11, His12, Ser13, and Asn14. These surfaces on the ligand could be essential for the subtype-specific recognition of the receptor. PMID- 9398299 TI - Tyrosine and tryptophan structure markers in hemoglobin ultraviolet resonance Raman spectra: mode assignments via subunit-specific isotope labeling of recombinant protein. AB - Phenyl-deuterated tyrosine (Tyr-d4) and indole-deuterated tryptophan (Trp-d5) have been selectively incorporated into hemoglobin (Hb) by expressing the gene in auxotrophic strains of Escherichia coli. Ultraviolet resonance Raman (UVRR) spectra, using 229-nm excitation, show that difference features characteristic of the Hb quaternary R --> T transition are not perturbed by the incorporation of the isotopes. All the UVRR bands between 800 and 1700 cm-1 are assigned to either Tyr or Trp except for the 1511 cm-1 band, which had been thought to arise from the Trp 2 x W18 overtone. This band does not shift upon Trp or Tyr labeling but does shift 5 cm-1 in D2O, suggesting assignment to a histidine (His) residue. Its intensification in the T-state is consistent with His protonation. The alpha- and beta-subunits were selectively labeled, by reconstitution of labeled subunits with unlabeled subunits, to make isotope hybrids. Selective Tyr labeling identified the alpha subunits as the locus of the Y8a upshift observed in Hb, supporting the previous inference that this shift is associated with the T-state H-bond involving the interfacial Tyr alpha42 [Rodgers, Su, Subramaniam, & Spiro (1992) J. Am. Chem. Soc. 114, 3697]. Selective Trp labeling showed the Trp alpha14 contributions to the T - R difference spectrum to be negligible and confirmed Trp beta37 as the locus of the W3 difference signal, and probably of the remaining Trp signals as well. The observed downshift of W17 and upshift of Wd5 in the T-state are consistent with a stronger T-state H-bond between Trp beta37 and Asp alpha94; the resulting excitation profile red shift accounts for the dominance of the Trp beta37 contribution to the T - R difference UVRR spectrum. PMID- 9398300 TI - Flavin conformational changes in the catalytic cycle of p-hydroxybenzoate hydroxylase substituted with 6-azido- and 6-aminoflavin adenine dinucleotide. AB - Crystallographic studies have demonstrated two flavin conformations for p hydroxybenzoate hydroxylase (PHBH) [Gatti, D. L., Palfey, B. A. , Lah, M. S., Entsch, B., Massey, V., Ballou, D. P., & Ludwig, M. L. (1994) Science 266, 110 114. Schreuder, H. A., Mattevi, A., Obmolova, G., Kalk, K. H., Hol, W. G. J., van der Bolt, F. J. T., & van Berkel, W. J. H. (1994) Biochemistry 33, 10161-10170]. The isoalloxazine ring system of one conformation (the "out" conformation) is significantly more exposed to solvent and is not in position for necessary catalytic reactions, but when the natural substrate is bound to the enzyme, the isoalloxazine is in the correct position (the "in" conformation) for its chemical function. In this study, several aspects of the function of the conformational change in catalysis were explored using the wild-type and Tyr222Phe forms of PHBH substituted with 6-azido FAD. This flavin served as both a spectral probe and a photolabel. The enzyme containing 6-azido FAD was a relatively effective catalyst for the hydroxylation of p-hydroxybenzoate. However, the intermediate reduced 6 azido enzyme was chemically unstable, and a small fraction converted to 6-amino PHBH by the elimination of N2 during each catalytic cycle. The reduction of 6 azido FAD PHBH by NADPH was almost as fast as the reduction of the natural enzyme. The characteristic spectral change caused by NADPH binding prior to hydride transfer strongly suggests that flavin movement from the "in" to the "out" conformation precedes flavin reduction. Irradiation of 6-azido PHBH with visible light covalently labeled proline 293, an active site residue, under conditions in which the flavin adopted the "in" conformation, while no protein labeling occurred under conditions in which the flavin was "out". The labeled protein exchanged substrate and was reduced by NADPH much more slowly than before photolysis. It is therefore concluded that isoalloxazine movement is required for pyridine nucleotide to gain access to the active site and for the exchange of aromatic ligands. PMID- 9398301 TI - Inactivation of 4-oxalocrotonate tautomerase by 2-oxo-3-pentynoate. AB - The compound, 2-oxo-3-pentynoate, has been synthesized and tested as an inhibitor of the enzyme 4-oxalocrotonate tautomerase. The enzyme is rapidly and irreversibly inactivated by the acetylenic product analogue in a time-dependent fashion. The enzyme displays saturation kinetics and is protected from inactivation by the presence of substrate. These observations are consistent with inactivation taking place at the active site. Partial reactivation ( approximately 18%) occurs by incubating the inactivated enzyme with 10 mM hydroxylamine (pH 7.3). The partition ratio, determined to be approximately 0.4, suggests that the inactivation of 4-OT by 2-oxo-3-pentynoate shows half-of-the sites stoichiometry. The same phenomenon is observed in the inactivation of 4-OT by 3-bromopyruvate and can be explained by examination of the crystal structure. Mass spectral analysis shows that a single residue is modified on the enzyme which has been localized to the nine residue amino-terminal fragment Pro-1 to Glu 9. It can be reasonably concluded that Pro-1 is the site of covalent attachment. Inactivation of 4-OT can occur by either a Michael addition of 4-OT to C-4 of 2 oxo-3-pentynoate or by the enzyme-catalyzed rearrangement of 2-oxo-3-pentynoate to an allene derivative which alkylates Pro-1. These results provide the foundation for the use of 2-oxo-3-pentynoate in future mechanistic studies and as a ligand in an inactivated 4-OT complex that can be studied by X-ray crystallography. Finally, 2-oxo-3-pentynoate is an acetylene analogue of a variety of 2-oxo acids and as such may have general utility as an inhibitor of reactions that bind and process these compounds. PMID- 9398302 TI - Participation of ADP dissociation in the rate-determining step in cAMP-dependent protein kinase. AB - Pre-steady-state kinetic analyses of the catalytic subunit of cAMP-dependent protein kinase showed that the rate constant for phosphoryl transfer is fast and either the release of one or both of the products or a conformational change controls turnover [Grant, B., & Adams, J. A. (1996) Biochemistry 35, 2022-2029]. To determine which step or steps control turnover in the wild-type enzyme, we used a catalytic trapping technique to measure directly the dissociation rate constant for ADP. The phosphorylation of two peptide substrates, LRRASLG and GRTGRRNSI, was monitored using a rapid quench flow technique under conditions where saturating concentrations of ADP were preequilibrated with the enzyme before excess ATP and one of the substrates were added to trap the free enzyme and to start the phosphorylation reaction. Under ADP preequilibration conditions, no 'burst' phase was observed, and although the rate of linear, steady-state turnover was unaffected, the net production of phosphopeptide lagged behind the non-preequilibrated control. This phenomenon occurs due to the slow release of the product, and kinetic modeling suggests that this effect can be explained if the dissociation rate constant for ADP is 24 s-1 and solely limits turnover (kcat = 23 s-1) for the phosphorylation of LRRASLG. Using GRTGRRNSI, the dissociation rate constant for ADP is 35 s-1 and limits turnover (kcat = 29 s-1) if the reaction is initiated by the addition of enzyme. Under preequilibration conditions with either ATP or GRTGRRNSI, turnover is approximately 50% lower, suggesting that ADP release may partially control this parameter. This preequilibration effect can be explained by slowly interconverting enzyme forms with specific peptide-induced turnover properties. These studies indicate that ADP release is an essential rate-limiting component for turnover but also suggests that other factors contribute subtly when the structure of the substrate is altered. PMID- 9398303 TI - Changes in protonation associated with substrate binding and Cob(I)alamin formation in cobalamin-dependent methionine synthase. AB - Methionine synthase catalyzes the transfer of a methyl group from methylcobalamin enzyme to homocysteine, generating methionine and cob(I)alamin enzyme, and then from methyltetrahydrofolate to cob(I)alamin enzyme, generating tetrahydrofolate and regenerating the methylcobalamin enzyme. The reactions catalyzed by methionine synthase require deprotonation of the substrate, homocysteine, and protonation of the product tetrahydrofolate, with no net change in proton stoichiometry for a complete turnover cycle. In addition, formation of the intermediate cob(I)alamin enzyme requires a change in the cobalt ligand geometry from 6-coordinate to 4-coordinate, and this rearrangement may require the transient protonation of protein residues to stabilize the cob(I)alamin enzyme. In the E. coli enzyme, the lower face of the methylcobalamin cofactor is coordinated by histidine 759, which is hydrogen bonded to aspartate 757 and then to serine 810, forming a "ligand triad". It has previously been shown that reduction of cob(II)alamin enzyme to cob(I)alamin is associated with the uptake of a proton from solution, and it has been postulated that this proton resides within the His759-Asp757 pair. Cob(I)alamin can also be generated by demethylation of methylcobalamin enzyme by homocysteine; it was not known whether this mode of cob(I)alamin formation was associated with proton uptake. In this paper, we use equilibrium titrations and kinetic analyses in the presence of the pH indicator dye phenol red, along with studies of the pH dependence of oxidation/reduction equilibria, to identify and characterize mechanistic steps associated with proton uptake and release in both the turnover and reactivation of the enzyme. We confirm that cob(I)alamin formation by reduction of cob(II)alamin enzyme is associated with proton uptake and show that mutation of Asp757 to Glu abolishes the pH dependence of this reduction. Demethylation of methylcobalamin enzyme also leads to cob(I)alamin formation and is also shown to be associated with proton uptake. By observing pre-steady-state reactions with homocysteine and methyltetrahydrofolate in the presence of phenol red, we show that this proton uptake occurs at a rate that is equal to the rate of formation of the cob(I)alamin enzyme. In addition, we show that binding of homocysteine to the enzyme results in the rapid release of a proton, presumably the homocysteine thiol proton. In contrast, binding methyltetrahydrofolate to the enzyme does not result in proton uptake, suggesting that the proton destined for the product tetrahydrofolate is already present on the free methylcobalamin enzyme. PMID- 9398304 TI - Cobalamin-dependent methionine synthase from Escherichia coli: involvement of zinc in homocysteine activation. AB - Methionine synthase (MetH) is a modular protein with at least four distinct regions; amino acids 2-353 comprise a region responsible for binding and activation of homocysteine, amino acids 345-649 are thought to be involved in the binding and activation of methyltetrahydrofolate, amino acids 650-896 are responsible for binding of the prosthetic group methylcobalamin, and amino acids 897-1227 are involved in binding adensylmethionine and are required for reductive activation of enzyme in the cob(II)alamin form. Previous studies have shown that mutations of Cys310 or Cys311 to either alanine or serine result in loss of all detectable catalytic activity. These mutant proteins retain the ability to catalyze methyl transfer from methyltetrahydrofolate to exogenous cob(I)alamin, but have lost the ability to transfer methyl groups from exogenous methylcobalamin to homocysteine [Goulding, C. W., Postigo, D., and Matthews, R. G. (1997) Biochemistry 36, 8082-8091]. We now demonstrate that both MetH holoenzyme and a truncated MetH(2-649) protein, which lacks a cobalamin prosthetic group, contain 0.9 equiv of zinc, while the Cys310Ser and Cys311Ser mutant proteins contain less than 0.05 equiv of zinc. Addition of l-homocysteine to MetH(2-649) is accompanied by release of 1 equiv of protons/mol of protein, while addition of l-homocysteine to the Cys310Ser and Cys311Ser mutant truncated proteins does not result in proton release. The Cys310Ala and Cys311Ala mutant methylcobalamin holoenzymes have completely lost the ability to transfer the methyl group from methylcobalamin to homocysteine, suggesting that zinc is required for this reaction. Further evidence that zinc is required for catalytic activity comes from experiments in which the zinc is removed from MetH(2-1227). Removal of zinc from methylated wild-type holoenzyme by treatment with methyl methanethiolsulfonate and then with dithiothreitol and EDTA results in loss of the ability of the protein to catalyze methyl transfer from methyltetrahydrofolate to homocysteine. Reconstitution of the zinc-depleted holoenzyme results in incorporation of 0.4 equiv of zinc/mol of protein and partial restoration of the ability of the protein to catalyze homocysteine methylation. PMID- 9398305 TI - Glutamate and aspartate as proton shuttles in mutants of carbonic anhydrase. AB - Maximal turnover rates for the hydration of CO2 and the depletion of 18O from CO2 catalyzed by carbonic anhydrase III (CA III) and carbonic anhydrase V (CA V) are limited by proton transfer involving zinc-bound water or hydroxide in the active site. We have investigated the capacity of glutamic and aspartic acids at position 64 in human CA III and murine CA V to act as proton shuttles in this pathway. The distance from the Calpha of position 64 to the zinc is near 9.5 A in the crystal structures of both CA III and CA V. Rates of intramolecular proton transfer between these proton shuttle groups and the zinc-bound water molecule were estimated as the predominant rate-contributing step in the catalytic turnover kcat in the hydration of CO2 measured by stopped flow and in the 18O exchange between CO2 and water measured by mass spectrometry. We found that both glutamate and aspartate residues at position 64 are efficient proton shuttles in HCA III. The rate constant for intramolecular proton transfer from either residue to zinc-bound hydroxide is 4 x 10(4) s-1, about 20-fold greater than that of the wild type which has lysine at position 64. When the active site residue Phe 198 in human CA III was replaced with Leu, measurement of catalysis showed that Glu 64 retained but Asp 64 lost its capacity to act as a proton shuttle. These observations were supported in studies of catalysis by murine CA V which contains Leu 198; here again, Glu 64 acted as a proton shuttle, but Asp 64 did not. Phe 198 in HCA III is thus a significant factor in the capacity of the active site to sustain proton transfer, possibly through its stabilization of hydrogen-bonded water bridges that enhance proton translocation from both Glu and Asp at position 64 to the zinc-bound hydroxide. PMID- 9398306 TI - Feed-back inhibition of oxidative stress by oxidized lipid/amino acid reaction products. AB - Three oxidized lipid/amino acid reaction products (OLAARPs): 1-methyl-4-pentyl 1,4-dihydropyridine-3,5-dicarbaldehyde, 1-(5-amino-1-carboxypentyl)pyrrole, and N (carbobenzyloxy)-1(3)-[1-(formylmethyl)hexyl]-l-histidine dihydrate, were prepared and tested for antioxidative activity in a microsomal system in order to investigate the effect that OLAARP formation may be playing in the oxidative stress process. The microsomal system consisted of freshly prepared trout muscle microsomes, which were oxidized in the presence of 5 microM Cu2+, 1 mM Fe3+/5 mM ascorbate, or 1 mM Cu2+/10 mM H2O2, and the compound to be tested as antioxidant added at 50 microM. At different periods of time, samples were tested for lipid peroxidation, assessed by the formation of thiobarbituric acid reactive substances (TBARS), and protein damage, which was evaluated by the formation of protein carbonyls and amino acid analysis. The three OLAARPs and butylated hydroxytoluene significantly (p < 0.05) protected against lipid peroxidation and protein damage for the three systems assayed. On the contrary, neither the amino acids used in the preparation of OLAARPs nor alpha-tocopherol, mannitol, aminoguanidine, or 4, 5-dihydroxy-1,3-benzenedisulfonic acid exhibited this constant protection. Because OLAARPs were produced at inhibitory levels during microsomal lipid peroxidation, these results suggest that OLAARP formation may be an antioxidative defense mechanism by which oxidative stress is feed-back inhibited, delaying the damage caused by reactive oxygen species. PMID- 9398307 TI - Molecular mechanism of regulation of the pyruvate dehydrogenase complex from E. coli. AB - The pyruvate dehydrogenase multienzyme complex from E. coli shows a sigmoidal dependency of the reaction rate on the substrate concentration when product formation is followed in the presence of physiological concentrations of the cofactor thiamin diphosphate. To elucidate the molecular mechanism of this regulation, the influence of the substrate pyruvate on the coenzyme-protein interaction has been investigated using several coenzyme analogues. The observed binding constants of all coenzymatically active analogues are increased in the presence of the substrate pyruvate, whereas those of all coenzymatically inactive analogues are not altered in the presence of pyruvate. This points to an increased binding affinity of a reaction-intermediate-coenzyme complex to the protein. Since cofactor binding and dissociation at physiological concentrations of thiamin diphosphate are slow compared to the catalytic reaction, a slow transition to the active state of the enzyme occurs. After lowering the pyruvate concentration, the opposite effect, a dissociation of the thiamin diphosphate from the enzyme is observed. This slow substrate dependent enhancement of cofactor binding enables efficient regulation of the pyruvate dehydrogenase complex by its substrate pyruvate. PMID- 9398308 TI - Histidine --> carboxamide ligand substitutions in the zinc binding site of carbonic anhydrase II alter metal coordination geometry but retain catalytic activity. AB - The catalytic zinc ion of human carbonic anhydrase II (CAII) is coordinated by three histidine ligands (H94, H96, and H119) and a hydroxide ion with tetrahedral geometry. Structural and functional analysis of variants in which the zinc ligands H94 and H119 are substituted with asparagine and glutamine, and comparison with results obtained with aspartate and glutamate substitutions indicate that the neutral ligand field provided by the protein optimizes the electrostatic environment for the catalytic function of the metal ion, including stabilization of bound anions. This is demonstrated by catalytic activity measurements for ester hydrolysis and CO2 hydration, as well as sulfonamide inhibitor affinity assays. High-resolution X-ray crystal structure determinations of H94N, H119N, and H119Q CAIIs reveal that the engineered carboxamide side chains coordinate to zinc with optimal stereochemistry. However, zinc coordination geometry remains tetrahedral only in H119Q CAII. Metal geometry changes to trigonal bipyramidal in H119N CAII due to the addition of a second water molecule to the zinc coordination polyhedron and also in H94N CAII due to the displacement of zinc-bound hydroxide by the bidentate coordination of a Tris molecule. Possibly, the bulky histidine imidazole ligands of the native enzyme play a role in disfavoring trigonal bipyramidal coordination geometry for zinc. Protein-metal affinity is significantly compromised by all histidine --> carboxamide ligand substitutions. Diminished affinity may result from significant movements (up to 1 A) of the metal ion from its position in the wild-type enzyme, as well as the associated, minor conformational changes of metal ligands and their neighboring residues. PMID- 9398309 TI - Evidence for a functional role of the dynamics of glycine-121 of Escherichia coli dihydrofolate reductase obtained from kinetic analysis of a site-directed mutant. AB - Two-dimensional heteronuclear (1H-15N) nuclear magnetic relaxation studies of dihydrofolate reductase (DHFR) from Escherichia coli have demonstrated that glycine-121 which is 19 A from the catalytic center of the enzyme has large amplitude backbone motions on the nanosecond time scale [Epstein, D. M., Benkovic, S. J., and Wright, P. E. (1995) Biochemistry 34, 11037-11048]. In order to probe the dynamic-function relationships of this residue, we constructed a mutant enzyme in which this glycine was changed to valine. Equilibrium binding studies indicated that the Val-121 mutant retained wild-type binding properties with respect to dihydrofolate and tetrahydrofolate; however, binding to NADPH and NADP+ was decreased by 40-fold and 2-fold, respectively, relative to wild-type DHFR. Single-turnover experiments indicated that hydride transfer was reduced by 200-fold to a rate of 1.3 s-1 and was the rate-limiting step in the steady state. Interestingly, pre-steady-state kinetic analysis of the Val-121 mutant revealed a conformational change which preceded chemistry that occurred at a rate of 3.5 s 1. If this step exists in the kinetic mechanism of the wild-type enzyme, then it would be predicted to occur at a rate of approximately 2000 s-1. Glycine-121 was also changed to alanine, serine, leucine, and proline. While the Ala-121 and Ser 121 mutants behaved similar to wild-type DHFR, the Leu-121 and Pro-121 mutants behaved like Val-121 DHFR in that hydride transfer was the rate-limiting step in the steady state and a conformational change preceding chemistry was observed. Finally, insertion of a glycine or valine between amino acids 121 and 122 produced mutant enzymes with properties similar to wild-type or Val-121 DHFRs, respectively. Taken together, these results provide compelling evidence for dynamic coupling of a remote residue to kinetic events at the active site of DHFR. PMID- 9398310 TI - Modification of aldose reductase by S-nitrosoglutathione. AB - Kinetic and structural changes in recombinant human aldose reductase (AR) due to modification by S-nitrosoglutathione (GSNO) were investigated. Incubation of the enzyme with 10-50 microM GSNO led to a time- and concentration-dependent inactivation of the enzyme, with a second-order rate constant of 0.087 +/- 0.009 M-1 min-1. However, upon exhaustive modification, 30-40% of the enzyme activity was retained. The non-inactivated enzyme displayed a 2-3-fold change in Km for NADPH and Km fordl-glyceraldehyde, whereas the Km for the lipid peroxidation product, 4-hydroxy-2-trans nonenal (HNE), was comparable to that of the untreated enzyme. The residual activity of the enzyme after GSNO treatment was less sensitive to inhibition by the active site inhibitor sorbinil or to activation by sulfate. Significantly higher catalytic activity was retained when the enzyme was modified in the presence of NADPH, suggesting relatively low reactivity of the E NADPH complex with GSNO. The modification site was identified using site-directed mutants in which each of the solvent-exposed cysteines of the enzyme was replaced individually by serine. The mutant C298S was insensitive to GSNO, whereas the sensitivity of the mutants C303S and C80S was comparable to that of the wild-type enzyme. Electrospray ionization mass spectroscopy of the GSNO-modified enzyme revealed a major modified species (70% of the protein) with a molecular mass that was 306 Da higher than that of the untreated enzyme, which is consistent with the addition of a single glutathione molecule to the enzyme. The remaining 30% of the protein displayed a molecular mass that was not significantly different from that of the native enzyme. No nitrosated forms of the enzyme were observed. These results suggest that inactivation of AR by GSNO is due to the selective formation of a single mixed disulfide between glutathione and Cys-298 located at the NADP(H)-binding site of the enzyme. PMID- 9398311 TI - General acid/base catalysis in the active site of Escherichia coli thioredoxin. AB - Enzymic catalysts of thiol:disulfide oxidoreduction contain two cysteine residues in their active sites. Another common residue is an aspartate (or glutamate), the role of which has been unclear. Escherichia coli thioredoxin (Trx) is the best characterized thiol:disulfide oxidoreductase, and in Trx these three active-site residues are Cys32, Cys35, and Asp26. Structural analyses had indicated that the carboxylate of Asp26 is positioned properly for the deprotonation of the thiol of Cys35, which would facilitate its attack on Cys32 in enzyme-substrate mixed disulfides. Here, Asp26 of Trx was replaced with isologous asparagine and leucine residues. D26N Trx and D26L Trx are reduced and oxidized more slowly than is wild type Trx during catalysis by E.coli thioredoxin reductase. Stopped-flow spectroscopy demonstrated that the cleavage of the mixed disulfide between Trx and a substrate is slower in the D26N and D26L enzymes. Buffers increase the rate of mixed disulfide cleavage in these variants but not in wild-type Trx. These results indicate that Asp26 serves as an acid/base in the oxidation/reduction reactions catalyzed by Trx. Specifically, Asp26 protonates (during substrate oxidation) or deprotonates (during substrate reduction) the thiol of Cys35. A similar role is likely filled by the analogous aspartate (or glutamate) residue in protein disulfide isomerase, DsbA, and other thiol:disulfide oxidoreductases. Moreover, these results provide the first evidence for general acid/base catalysis in a thiol:disulfide interchange reaction. PMID- 9398312 TI - Steady-state kinetics and inhibitor binding of 3-deoxy-D-arabino-heptulosonate-7 phosphate synthase (tryptophan sensitive) from Escherichia coli. AB - The tryptophan-inhibited 3-deoxy-d-arabino-heptulosonate-7-phosphate synthase [DAHPS(Trp)] of Escherichia coli was analyzed with respect to steady-state kinetics and tryptophan binding. DAHPS(Trp) is one of three differentially regulated isoforms that catalyze the first step of aromatic biosynthesis, the condensation of phosphoenolpyruvate and erythrose-4-phosphate to form 3-deoxy-D arabino-heptulosonate-7-phosphate. The DAHP synthase isozymes are metalloproteins, being activated in vitro by a variety of divalent metals. Both catalytic activity and substrate affinity are dependent on the species of activating metal ion. We report here kinetic and binding studies of metal homogeneous (Mn2+-activated) DAHPS(Trp). The homodimeric enzyme had an apparent kcat of 21 s-1 and displayed sigmoidal kinetics with respect to both substrates. The S0.5 was 35 microM for erythrose-4-phosphate and 5.3 microM for phosphoenolpyruvate. Equilibrium binding studies with radiolabeled tryptophan demonstrated two independent inhibitor binding sites per enzyme dimer, with KdTrp of 1 microM. L-Tryptophan binding decreased kcat, increased affinity for both substrates, decreased positive homotropic cooperativity for both substrates and activated the enzyme at low concentrations of erythrose-4-phosphate. The results suggest an inhibition mechanism analogous to system C5 hyperbolic mixed-type inhibition with respect to erythrose-4-phosphate and partial noncompetitive inhibition with respect to phosphoenolpyruvate. PMID- 9398313 TI - Use of thiouredopyrenetrisulfonate photochemistry for driving electron transfer reactions in aqueous solutions. AB - Photoexcitation of 1-thiouredopyrene-3,6,8-trisulfonic adducts (TUPS) of amino acids by the third harmonic frequency of a Nd:YAG laser (355 nm) generates the triplet state of the dye with high quantum efficiency. Relaxation of the triplet proceeds in anaerobiosis with a half decay time of 0.5 ms. The relaxation rate increases 100-fold in the presence of dioxygen. A radiative transition between the triplet and the ground state of the dye results in phosphorescent emission centered at 658 nm. The excited state of TUPS, being a strong reductant, can donate its electron to a variety of acceptors. Transient absorption spectroscopy was used to directly measure the photoinduced electron transfer from the excited dye to rhodamine B (RB) and cytochrome c. The reaction with RB was followed by monitoring the oxidation of the triplet state of TUPS at 487 nm (epsilon = 25 000 +/- 5 000 M-1 cm-1) or the reduction of RB at 553 nm. The second order rate constant for the reaction was found to be (2.5 +/- 0.2) x 10(9) M-1 s-1, a value compatible with that for diffusion controlled reactions. When directed to cytochrome c the photoinduced perturbation causes rapid reduction of the protein's heme group, seen as a monophasic increase of absorbance at 550 nm. The combination of appropriate redox properties with the capability of covalent protein modification makes the dye useful for initiation and analysis of electron transfer reactions in chemical and biological systems. PMID- 9398314 TI - Photoinduced electron transfer in singly labeled thiouredopyrenetrisulfonate cytochrome c derivatives. AB - A novel method for initiation of intramolecular electron transfer reactions in cytochrome c is reported. The method is based on photoexcitation of covalently attached thiouredopyrenetrisulfonate (TUPS) by the third harmonic frequency of a Nd:YAG laser (355 nm), the reaction that generates the low-potential triplet state of the dye with high quantum efficiency. TUPS derivatives of horse heart cytochrome c singly labeled at specific lysine residues were prepared and purified to homogeneity by ion-exchange high-pressure liquid chromatography. Eight derivatives were characterized by determination of the location of the modification, reduction potentials, and measurement of enzymatic activity with cytochrome oxidase. Transient absorption spectroscopy was used to directly measure the rate constants for the electron transfer reaction from the photoexcited triplet state of TUPS to the oxidized heme group and the back reaction from the ferrous heme to the oxidized dye. For all singly labeled derivatives, the rate constants for heme reduction were 1 or 2 orders of magnitude larger than for its reoxidation, consistent with the greater thermodynamic driving force for the oxidation reaction. Analysis of the variation of electron-transfer rates with the distance separating the dye and the heme reveals a value of coupling decay constant (beta) of 0.46 A-1. Rapid and effective photoreduction of cytochrome c makes it a useful tool for fast initiation of electron transfer in the reductive direction within complexes of cytochrome c with other redox proteins. PMID- 9398315 TI - Cooperativity and regulation of scallop myosin and myosin fragments. AB - Scallop heavy meromyosin (HMM) preparation obtained by a new improved method showed a Mg-ATPase activity that was activated 15-fold by calcium. The ATPase activity depended on ionic strength and reached maximum at 0.1 M without altering calcium sensitivity. The highly regulated HMM and myosin preparations showed cooperative properties not seen with unregulated subfragment 1 (S1). ATPase activity of myosin and HMM increased steeply with calcium concentration, yielding Hill coefficients about 3 and 4, respectively. Calcium binding by HMM and myosin became cooperative in the presence of ADP, AMP-PNP, or ADP.Vi yielding Hill coefficients of 1.8 and 2.8, respectively. Binding of calcium by HMM in the presence of ATP was also cooperative at physiological ionic strength, whereas at low ionic strength the data fit best to a simple binding curve. In contrast, calcium binding by unregulated S1 followed a normal binding curve and was not affected by the presence of nucleotide analogues. Calcium decreased the affinity of ADP and ADP-PNP to myosin and HMM, but had no effect on the nucleotide binding to S1. The results indicate that communication between the nucleotide and calcium binding sites requires the presence of two heads and exists only in the "off" state. We propose that in the presence of calcium, interaction between the two heads is disrupted and they act independently. PMID- 9398316 TI - Alignment of fibrillin molecules in elastic microfibrils is defined by transglutaminase-derived cross-links. AB - Microfibrils were extracted from human amnion in the form of a beaded filament and analyzed for the presence of transglutaminase-derived cross-links using acrylonitrile derivatization. The cross-link structure was isolated from protease hydrolysates of beaded filaments and identified as a phenylthiocarbamyl amino acid derivative by comparison to a standard. Acid hydrolysis of the isolated cross-link gave the expected lysine and glutamic acid in a 1:1 ratio. The beaded filaments were also treated with trypsin to produce a fraction that contained the bead structure and a fraction containing fragments of the interbead filaments. Cross-links were detected in the interbead filaments but not in the beads. A large tryptic peptide that contained a cross-link was isolated and sequenced. The two amino acid sequences obtained identified both of the cross-linked molecules as fibrillin-1 and enabled the approximate localization of the cross-link sites within the molecule. The locations of cross-link sites on two adjacent molecules fixed the relative positions of fibrillin monomers within the microfibrils, providing insight into the spatial organization of fibrillin within the elastic microfibrils. PMID- 9398317 TI - Probing the effects of calcium on gelsolin. AB - Gelsolin is a calcium-regulated actin severing and capping protein that binds two calcium ions and has three sites for actin; two recognize monomeric actin and one attaches to the sides of filaments. It contains six repeating sequence segments (G1-6). Here, we have analyzed the effects of calcium ions on (i) limited proteolysis of bacterially expressed human gelsolin by plasmin and (ii) dynamic light scattering and circular dichroism of gelsolin and various of its subdomains. Following cleavage of gelsolin in the absence of calcium between Lys150 and His151 (the junction between G1 and G2), the molecule does not fall apart, nor does it bind actin without added calcium. This same molecule can be reconstituted by mixing an excess of G1 with G2-6 in EGTA. The noncovalently linked form of gelsolin shows three actin binding sites in calcium and requires 3 microM calcium for 50% activation of actin binding. Measurements of light scattering and circular dichroism revealed structural changes in response to calcium for intact gelsolin and a number of its actin-binding subdomains. Many of these changes occurred at calcium concentrations below 100 nM. These results are discussed in relation to the calcium control of gelsolin function and its three dimensional structure (Burtnick et al.(1997) Cell 90, 661-670). Nanomolar concentrations of calcium initiate the unlatching of structural constraints that maintain the inaccessibility of the actin binding sites, but actin binding occurs only after additional micromolar calcium sites in both the N-terminal and C terminal halves of the molecule are occupied. PMID- 9398318 TI - Identification of acetylcholine receptor channel-lining residues in the M1 segment of the beta-subunit. AB - The substituted cysteine accessibility method (SCAM) was applied to the first membrane-spanning segment (M1) of the mouse-muscle acetylcholine (ACh) receptor beta subunit. One at a time, each residue from betaR219 to betaP247, except betaC233, was mutated to Cys, and the mutant beta subunits were expressed together with wild-type alpha, gamma, and delta in Xenopus oocytes. All 28 mutants yielded functional receptors. The accessibility of the substituted Cys to the methanethiosulfonate (MTS) derivatives, MTS ethylammonium (MTSEA), MTS ethyltrimethylammonium (MTSET), and MTS ethylsulfonate (MTSES), added extracellularly in the absence or the presence of ACh, was inferred from their irreversible effects on ACh-induced current. Three consecutive residues close to the extracellular end of M1, betaF224C, betaY225C, and betaL226C, reacted both in the absence and presence of ACh, and one deeper residue, betaV229C reacted only in the presence of ACh. betaV229C also reacted with 2-aminoethyl-2 aminoethanethiosulfonate (AEAETS) and with 2-hydroxyethyl MTS (MTSEH). The rate constants for the reactions of betaV229C with MTSEA, which permeates the open channel, and with MTSEH, which is uncharged, were independent of membrane potential. The rate constant for the reaction of the doubly positively charged AEAETS, however, was dependent on membrane potential, consistent with the exposure of betaV229C in the open channel. The N-terminal third of betaM1, like that of alphaM1, contributes to the lining of the channel and undergoes structural changes during gating. PMID- 9398319 TI - Orientation in lipid bilayers of a synthetic peptide representing the C-terminus of the A1 domain of shiga toxin. A polarized ATR-FTIR study. AB - The interaction of a synthetic peptide representing the C-terminal 27 amino acids of the A1 domain of Shiga toxin (residues 220-246) with acidic phospholipid model membranes was characterized by FTIR spectroscopy. This peptide resembles a signal sequence and may mediate the translocation of the catalytic A1 chain of Shiga toxin to the cytoplasm following its retrograde transport to the lumenal compartment of the endoplasmic reticulum (ER). At pH 7 and 5, the peptide underwent a conformational change from random coil to alpha-helix upon addition of negatively charged phospholipids. Examination of the amide II band in the spectrum of the complex at pH 7 and pH 5 showed that in both cases, the N-H groups in the peptide backbone are largely protected from H/D exchange. Using polarized attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) measurements, the orientation of the alpha-helical portion of the peptide was found to be almost perpendicular with respect to the membrane plane at pH 7. However, at pH 5.0-5.4, the alpha-helix axis was preferentially oriented parallel to the membrane plane. The results suggest that at the neutral pH of the ER lumen, the peptide may insert into the membrane, while at the lower pH levels present in earlier endocytic compartments, the peptide would be less likely to traverse the bilayer. In summary, this putative signal peptide may not be able to cause a significant translocation of the A1 domain of Shiga toxin to the cytosol until it reaches the neutral pH of the ER compartment. PMID- 9398320 TI - MAP4 is the in vivo substrate for CDC2 kinase in HeLa cells: identification of an M-phase specific and a cell cycle-independent phosphorylation site in MAP4. AB - We reported previously that cdc2 kinase decreased the microtubule-stabilizing ability of a major HeLa cell microtubule-associated protein, MAP4, by phosphorylation in vitro [Ookata, K., et al. (1995) J. Cell Biol. 128, 849-862]. An important question raised by this study is whether MAP4 is indeed phosphorylated by cdc2 kinase at mitosis in vivo. We present here evidence that cdc2 kinase is the major M-phase MAP4 kinase, and, further, we identify two phosphorylation sites within the proline-rich domain of MAP4. Metabolic 32P labeling showed the increased phosphorylation of MAP4 at mitosis. A specific inhibitor of cdc2 kinase, butyrolactone I, inhibited phosphorylation of MAP4 both in mitotic HeLa cells and in the mitotic HeLa cell extract. The phosphopeptide map analysis revealed the high similarity of in vivo labeled mitotic MAP4 to that phosphorylated by cdc2 kinase in vitro. Ser-696 and Ser-787, both of which lie within SPXK consensus sequences for cdc2 kinase, were identified as phosphorylation sites in the proline-rich region of MAP4 in vivo and in vitro. Immunoblotting with antibodies that recognize the phosphorylation state of Ser 696 or Ser-787 showed that Ser-787 in the SPSK sequence was specifically phosphorylated at mitosis while Ser-696 in the SPEK sequence was phosphorylated both at mitosis and in interphase. These results suggest that cdc2 kinase directly regulates microtubule dynamics at mitosis through phosphorylation of MAP4 at a number of sites, including Ser-787. PMID- 9398321 TI - Covalent attachment of ethidium to DNA results in enhanced topoisomerase II mediated DNA cleavage. AB - The classic DNA intercalator, ethidium, was used to probe the effects of (i) intercalation and (ii) covalent modification of the DNA on the catalytic activity of topoisomerase II. Ethidium bromide, which binds reversibly to DNA via intercalation, does not stimulate topoisomerase II-mediated DNA cleavage at concentrations up to 100 microM, indicating that the intercalative binding of this molecule to DNA is not sufficient to alter the activity of the enzyme. In contrast, covalent attachment of the photoreactive ethidium analog to DNA resulted in marked enhancement of topoisomerase II-mediated single- and double stranded DNA cleavage. This increase in DNA cleavage was observed at very low drug binding densities (<1 drug per 10-80 base pairs) which correspond to nanomolar concentrations, as compared with other topoisomerase II poisons such as etoposide or m-AMSA which require micromolar concentrations to elicit comparable DNA cleavage levels. Over the past decade, topoisomerase II has been an important target for a variety of clinically relevant anticancer agents due to the abilities of these agents to convert this enzyme to a cellular toxin resulting in an increase in the levels of enzyme-mediated DNA breaks. Modification of DNA by covalently attaching a DNA-targeting intercalating agent (i.e., ethidium bromide) resulted in a marked shift of the cleavage/religation equilibrium of the enzyme toward the cleaved state "poison" topoisomerase II as observed by the enhancement in single- and double-stranded cleavage; thus, key insight was gained into the mechanism(s) through which DNA binding agents may influence the catalytic properties of topoisomerase II. These data demonstrate that conversion of a reversible ethidium-DNA complex to an irreversible adduct results in the transformation of an ineffective intercalating drug into a potent topoisomerase II-targeted agent. Finally, they provide support for the recently proposed "positional poisoning model" for the actions of DNA lesions and anticancer drugs on the type II enzyme. PMID- 9398322 TI - Elongation properties of vaccinia virus RNA polymerase: pausing, slippage, 3' end addition, and termination site choice. AB - We have analyzed the elongation properties of vaccinia virus RNA polymerase during a single round of transcription in vitro. RNA-labeled ternary complexes were halted at a unique template position located upstream of a T-run (TTTTTTTTT) in the nontemplate strand; this element encodes an RNA signal for factor dependent transcription termination at distal sites on the template. The halted ternary complexes were purified and allowed to resume elongation under a variety of conditions. We found that the T-run constituted a strong elongation block, even at high nucleotide concentrations. The principal sites of pausing were at a C position situated two nucleotides upstream of the first T in the T-run and at the first three to four T positions within the T-run. There was relatively little pausing at the five downstream Ts. Intrinsic pausing was exacerbated at suboptimal nucleotide concentrations. Ternary complexes arrested by the T-run at 10 microM NTPs rapidly traversed the T-run when the NTP pool was increased to 1 mM. Limiting GTP (1 microM) resulted in polymerase stuttering at the 3' margin of the T-run, immediately prior to a templated G position; this generated a ladder of slippage synthesis products. We found that vaccinia ternary complexes remained intact after elongating to the very end of a linear DNA template and that such complexes catalyzed the addition of extra nucleotides to the 3' end of the RNA chain. The 3' end addition required much higher concentrations of NTPs than did templated chain elongation. Finally, we report that factor-dependent transcription termination by vaccinia RNA polymerase downstream of the T-run was affected by nucleotide concentration. Limiting UTP caused the polymerase to terminate at sites closer to the UUUUUNU termination signal. This is consistent with the kinetic coupling model for factor-dependent termination. PMID- 9398323 TI - DNA-Binding properties of the replication telomere protein. AB - The replication Telomere Protein, rTP, is a nuclear protein from the ciliate Euplotes crassus that appears to be a novel telomere replication factor. rTP shares extensive amino acid sequence identity with the two proteins that bind and protect the macronuclear telomeres from the ciliates Oxytricha and Euplotes. Since the most extended regions of conservation fall within the DNA-binding domains of the telomere-binding proteins, when rTP was first identified it was predicted to be another structural telomere-binding protein. However, subsequent research demonstrated that rTP transcripts accumulate only during DNA replication and the rTP protein localizes to the sites of DNA replication within Euplotes macronuclei. We have now expressed rTP in a heterologous expression system and have examined the DNA-binding properties of the recombinant protein. We show that rTP binds specifically to the G-strand of Euplotes telomeric DNA and hence has some of the same DNA-binding characteristics as the Euplotes and Oxytricha telomere-binding proteins. However, other aspects of rTP binding are unique. In particular, the protein exhibits a very high off-rate and can bind double stranded DNA as well as internal tracts of telomeric sequence. We conclude that rTP and the telomere-binding proteins are members of a class of proteins that have a conserved DNA-binding motif tailored to bind the G-strand of telomeric DNA. However, the unique DNA-binding characteristics of rTP indicate that the protein has evolved to fulfil a specialized role during telomere replication. PMID- 9398324 TI - A novel dCMP methylase by engineering thymidylate synthase. AB - X-ray crystal structures of binary complexes of dUMP or dCMP with the Lactobacillus caseiTS mutant N229D, a dCMP methylase, revealed that there is a steric clash between the 4-NH2 of dCMP and His 199, a residue which normally H bonds to the 4-O of dUMP but is not essential for activity. As a result, the cytosine moiety of dCMP is displaced from the active site and the catalytic thiol is moved from the C6 of the substrate about 0.5 A further than in the wild-type TS-dUMP complex. We reasoned that combining the N229D mutation with mutations at residue 199 which did not impinge on the 4-NH2 of dCMP should correct the displacements and further favor methylation of dCMP. We therefore prepared several TS N229D mutants and characterized their steady state kinetic parameters. TS H199A/N229D showed a 10(11) change in specificity for methylation of dCMP versus dUMP. The structures of TS H199A/N229D in complex with dCMP and dUMP confirmed that the position and orientation of bound dCMP closely approaches that of dUMP in wild-type TS, whereas dUMP was displaced from the optimal catalytic binding site. PMID- 9398325 TI - Human histone acetyltransferase GCN5 exists in a stable macromolecular complex lacking the adapter ADA2. AB - Acetylation of core histones is an important regulatory step in transcriptional activation from chromatin templates. The yeast transcriptional coactivator protein GCN5 was recently shown to be a nuclear histone acetyltransferase (HAT). Genetic and biochemical studies in yeast suggest that GCN5 functions with the adapter proteins ADA1, ADA2, ADA3, and ADA5 in a heteromeric complex. We have established conditions for chromatographic fractionation of HATs and ADA2 from human K562 erythroleukemia cells. Gel-filtration chromatography revealed two populations of GCN5 with Stokes' radii of 67 and 33 A, consistent with a large macromolecular complex and a monomer, respectively. The GCN5-related HAT, PCAF, was resolved as a stable complex with a Stokes' radius of 74 A. The HAT complexes were resistant to 0.3 M NaCl and DNase I. ADA2 was characterized by a Stokes' radius of 35 A, consistent with a monomer. Thus, in contrast to the stable GCN5 adapter complex in yeast, human GCN5 and ADA2 are not stably associated with each other. The implications of this result are discussed vis-a-vis the mechanism of recruitment of GCN5 to regulatory regions of genes. PMID- 9398326 TI - Short-chain phosphatidylinositol conformation and its relevance to phosphatidylinositol-specific phospholipase C. AB - The solution conformation of chiral diheptanoylphosphatidylinositol (D- and L inositol isomers) has been characterized by NMR spectroscopy. A positive NOE between the inositol C2 proton and an sn-3 glycerol CH2 proton has been observed in the D- but not in the L-inositol isomer of diheptanoylphosphatidylinositol (PI). Computer modeling using QUANTA constrained by this NOE and ring coupling constants suggests that the inositol ring is nearly parallel to the chain packing direction, leaving the phosphate ester accessible to attack by phosphatidylinositol-specific phospholipase C enzymes. In this model, the hydroxyl groups in the 2- and 6-positions of inositol form hydrogen bonds with the pro-R and ester oxygens, respectively. Chemical shifts and 13C spin-lattice relaxation times were also used to assess conformation and lipid dynamics in monomer and micelle states. The 13C T1's of inositol C2 and C6 in monomeric phosphatidylinositol were markedly less than for other inositol ring carbons. These results are consistent with the hydrogen bonds to the phosphate constraining the motions of C2 and C6. Diheptanoylphosphatidyl-2-O-methylinositol is a good inhibitor of PI-specific phospholipase C because it blocks the initial phosphotransferase step in PI hydrolysis. Introduction of the methyl group on the C-2 hydroxyl group lowers the CMC of the derivative compared to diheptanoylphosphatidylinositol. However, an NOE between an sn-3 glycerol proton and the inositol C2 proton constrains the orientation of the inositol ring with respect to the glycerol backbone in a conformation similar to diheptanoylphosphatidylinositol. Modeling of the 2-O-methylinositol derivative suggests that the methyl group blocks one side of the phosphate, consistent with the observation that nonspecific phospholipase C enzymes which are able to hydrolyze PI, albeit poorly, are unable to hydrolyze diheptanoylphosphatidyl-2-O methylinositol. PMID- 9398327 TI - Functional consequence of mutating conserved residues of the yeast farnesyl protein transferase beta-subunit Ram1(Dpr1). AB - Ras proteins, fungal mating pheromones, and other proteins terminating in the sequence CaaX (where C is Cys, a is any aliphatic amino acid, and X is the C terminal residue) are posttranslationally prenylated. Farnesyl-protein transferase (FPTase) transfers the farnesyl moiety of farnesyl pyrophosphate (FPP) to the thiol of the CaaX box cysteine in a reaction that requires Zn2+ and Mg2+. We have created mutations in conserved amino acids of the yeast Ram1 protein to identify residues important for Zn2+-dependent FPTase activity. Wild type and mutant Ram1 proteins were expressed as operon fusions in bacteria, and FPTase activity was measured. Mutations in conserved residues Glu256, His258, Asp307, Cys309, Asp360, and His363 reduce FPTase activity. Asp307, Cys309, and His363 correspond to the residues that have been shown to coordinate Zn2+ in mammalian FPTase. The H258N mutant enzyme exhibited an increased sensitivity to the Zn2+ chelator 1,10-phenanthroline, required higher concentrations of Zn2+ to restore activity to the apoenzyme, and had a 10-fold reduction in catalytic efficiency. The decreases in FPTase activity observed do not appear to be caused by major structural perturbations because the mutants were stably expressed and retained the ability to interact with Ram2p during purification. The FPTase activity of the mutants measured in vitro correlated well with their ability to complement the mating and growth defects of a ram1Delta strain in vivo. PMID- 9398328 TI - Structural studies of detergent-solubilized and vesicle-reconstituted low-density lipoprotein (LDL) receptor. AB - The low-density lipoprotein (LDL) receptor plays a key role in maintaining circulating and cellular cholesterol homeostasis. The LDL receptor is a transmembrane glycoprotein whose biochemical and genetic properties have been extensively studied notably by Brown, Goldstein and colleagues [Brown, M. S., & Goldstein, J. L., (1986) Science 232, 34-47]. However, few if any structural studies of the LDL receptor have been reported, and details of its secondary and tertiary structure are lacking. In an attempt to determine the low-resolution structure of the LDL receptor, we have purified the receptor from bovine adrenal cortices using modifications of the method of Schneider et al. [Schneider, W. J., Goldstein, J. L., & Brown, M. S. (1985) Methods in Enzymol.109, 405-417]. Using circular dichroism, the secondary structure of the detergent-solubilized bovine LDL receptor at 25 degrees C was shown to be 19% alpha-helix, 42% beta-sheet, and 39% random coil. Interestingly, the detergent-solubilized receptor appeared to be quite resistant to changes in secondary structure over the temperature range 10 90 degrees C, with only minor but reversible changes being observed. In contrast, a more pronounced unfolding of the detergent-solubilized receptor was observed in the presence of guanidinium hydrochloride. Using the complete sequence of the human LDL receptor, sequence analysis by the Chou-Fasman prediction algorithm showed quite good agreement with the experimentally determined secondary structure of the bovine LDL receptor at 25 degrees C. Finally, the purified, bovine LDL receptor was reconstituted into large unilamellar vesicles of egg yolk phosphatidylcholine using a procedure exploiting preformed vesicles and detergent dialysis. We showed previously using negative stain electron microscopy that reconstituted vesicles bind LDL. Now, using cryoelectron microscopy of frozen hydrated reconstituted vesicles evidence of an extended, stick-like morphology (length approximately 120 A) for the extracellular domain of the LDL receptor has been obtained. Successful purification of the receptor, its incorporation into single bilayer vesicles, and its direct visualization by cryoelectron microscopy pave the way for more detailed structural studies of the LDL receptor and the receptor-LDL complex. PMID- 9398329 TI - Suppression of phospholipase C beta, gamma, and delta families alters cell growth and phosphatidylinositol 4,5-bisphosphate levels. AB - Phosphatidylinositol-specific phospholipase C (PLC) activity reflects a summation of the activities of three families, beta, gamma, and delta, each of which is regulated differently. In order to understand the contribution of each family to cell proliferation signaling, expression of each family was suppressed by use of an inducible expression vector for antisense PLC sequences in a single cell line, FTO-2B rat hepatocytes. Activation of second messengers of PLC [diacylglycerol (DAG) and inositol 1,4,5-tris(phosphate) (IP3)] was dramatically reduced, providing a strategy for probing the consequences of PLC deficiency on cell function. Importantly, while one PLC family was suppressed, the other PLCs actively responded to specific stimuli, suggesting parallel and independent signaling pathways for each PLC family in FTO-2B cells. Selective suppression of each PLC family altered cell growth markedly and differentially. The rank order for suppression of cell growth by loss of a PLC family was gamma > delta > beta. Exploration of down-stream growth regulators revealed that loss of beta and gamma, but not delta, families was associated with markedly reduced basal ras and protein kinase C activity. Moreover, suppression of each of the three PLC families caused remarkably reduced basal and stimulated MAP kinase activities. Interestingly, cellular levels of PIP2 were increased and dramatically correlated with growth inhibition rate in the clones with suppressed PLC activity, suggesting that PIP2 itself can serve as a second messenger of cell growth regulation. PMID- 9398330 TI - Localization of the heme binding region in soluble guanylate cyclase. AB - Soluble guanylate cyclase (sGC) is a heterodimeric hemoprotein composed of alpha1 and beta1 subunits. sGC is activated by nitric oxide (NO) and therefore plays a central role in NO signal transduction. Activation of sGC by NO is believed to be mediated by the interaction between NO and the heme of sGC. Spectroscopic and kinetic studies have shown that the heme of sGC is in a unique environment. Characterization of the heme environment is critical to the understanding of the mechanism of NO activation. To approach this goal, the beta1 N-terminal fragment consisting of residues 1-385 [beta1(1-385)] of sGC was expressed in E. coli. beta1(1-385) was then purified to homogeneity in two steps by DEAE ion exchange and gel filtration chromatography. Purified beta1(1-385) was found to contain a stoichiometric amount of heme. The UV-visible spectrum of beta1(1-385) is almost identical to that of the native heterodimeric sGC purified from bovine lung. beta1(1-385) binds both NO and CO, leading to a shift in the Soret maximum from 431 nm to 398 and 423 nm, respectively. These spectral shifts are identical to those observed with heterodimeric sGC purified from bovine lung. These results suggest that the heme in the beta1(1-385) is similar to that in the heterodimeric sGC. Therefore, for the first time, the heme binding region of sGC has been unambiguously localized to the N-terminal region of the beta1 subunit. Our data also suggest that the N-terminal region of the beta1 subunit of sGC is itself sufficient for heme binding. PMID- 9398331 TI - A targeted library of small-molecule, tyrosine, and dual-specificity phosphatase inhibitors derived from a rational core design and random side chain variation. AB - Tyrosine phosphatases (PTPases) dephosphorylate phosphotyrosines while dual specificity phosphatases (DSPases) dephosphorylate contiguous and semicontiguous phosphothreonine and phosphotyrosine on cyclin dependent kinases and mitogen activated protein kinases. Consequently, PTPases and DSPases have a central role controlling signal transduction and cell cycle progression. Currently, there are few readily available potent inhibitors of PTPases or DSPases other than vanadate. Using a pharmacophore modeled on natural product inhibitors of phosphothreonine phosphatases, we generated a refined library of novel, phosphate free, small-molecule compounds synthesized by a parallel, solid-phase combinatorial-based approach. Among the initial 18 members of this targeted diversity library, we identified several inhibitors of DSPases: Cdc25A, -B, and C and the PTPase PTP1B. These compounds at 100 microM did not significantly inhibit the protein serine/threonine phosphatases PP1 and PP2A. Kinetic studies with two members of this library indicated competitive inhibition for Cdc25 DSPases and noncompetitive inhibition for PTP1B. Compound AC-alphaalpha69 had a Ki of approximately 10 microM for recombinant human Cdc25A, -B, and -C, and a Ki of 0.85 microM for the PTP1B. The marked differences in Cdc25 inhibition as compared to PTP1B inhibition seen with relatively modest chemical modifications in the modular side chains demonstrate the structurally demanding nature of the DSPase catalytic site distinct from the PTPase catalytic site. These results represent the first fundamental advance toward a readily modifiable pharmacophore for synthetic PTPase and DSPase inhibitors and illustrate the significant potential of a combinatorial-based strategy that supplements the rational design of a core structure by a randomized variation of peripheral substituents. PMID- 9398332 TI - Interaction of the cytoplasmic tail of CTLA-4 (CD152) with a clathrin-associated protein is negatively regulated by tyrosine phosphorylation. AB - CTLA-4 (CD152), high-avidity receptor for CD80 and CD86, is a powerful regulator of T cell activation. While CTLA-4 functions at the cell surface, it is primarily localized in intracellular vesicles and cycles to the cell surface. The CTLA-4 cytoplasmic domain contains sequences that direct its intracellular localization and regulate its signaling. Here we demonstrate that effector molecules involved in receptor trafficking and signaling interact with distinct, but overlapping, sequences in the CTLA-4 cytoplasmic domain. Using the yeast two-hybrid method, we demonstrate association of the mu2 subunit of AP-2, the clathrin-associated complex found in plasma membrane-associated coated pits, with the cytoplasmic tail of CTLA-4, but not CD28. The mu1 subunit of AP-1, found in Golgi-associated coated pits, associated with neither CTLA-4 nor CD28. Sequences required for interaction of mu2 and CTLA-4 were localized to residues, 161TTGVY in CTLA-4; this sequence is N-terminal to, but overlaps with, a previously identified SH2 binding motif, 165YVKM, involved in CTLA-4 signaling. Mu2 interacted preferentially with CTLA-4 when residue 165Y was nonphosphorylated, whereas a PI3 kinase SH2 domain interacted preferentially when 165Y was phosphorylated. In co transfection experiments, both tyrosine residues in the cytoplasmic tail of CTLA 4 (165Y and 182Y) were phosphorylated by the T lymphocyte-associated tyrosine kinase, p56lck. Thus, phosphorylation of CTLA-4 residue 165Y may reciprocally regulate signaling and trafficking of CTLA-4 by determining which effector molecules bind to its cytoplasmic tail. PMID- 9398333 TI - Site-directed mutagenesis of rubredoxin reveals the molecular basis of its electron transfer properties. AB - Rubredoxins contain a single non-heme iron atom coordinated by four cysteines. This iron is redox active and confers a role to these proteins in electron transfer chains. The structural features responsible for setting the values of the reduction potential and of the electron self-exchange rate constant have been probed by site-directed mutagenesis. Replacements of the highly conserved residues in positions 8, 10, and 11 (valine, glycine, and tyrosine, respectively) all lead to shifts of the reduction potential, up to 75 mV. These cannot be explained by simple considerations about the physicochemical properties of the substituting side chains but rather indicate that the value of the reduction potential is finely tuned by a variety of interactions. In contrast, the electron self exchange rate constant measured by nuclear magnetic resonance does not vary much, except when a charged residue is included in position 8 or 10, at the surface of the protein closest to the iron atom. Analysis of the data with a model for electrostatic interactions, including both monopolar and dipolar terms, indicates that the presence of a charge in this region not only increases the repulsion between molecules but also affects the electron transfer efficiency of the bimolecular complexes formed. The studies presented constitute a first step toward probing the structural elements modulating the reactivity of the FeS4 unit in a protein and defining the electron transfer active site(s) of rubredoxin. PMID- 9398334 TI - Role of beta87 Thr in the beta6 Val acceptor site during deoxy Hb S polymerization. AB - Three new Hb S variants containing beta87 Leu, Trp, or Asp instead of Thr were expressed in yeast in order to further define the role of the beta87 position in stability and polymerization of deoxy Hb S. Previous studies showed that hydrophobicity at beta85 Phe and beta88 Leu is critical for stabilization of hemoglobin. Results with the three Hb S beta87 variants, however, showed minimal differences in stability, suggesting that beta87 amino acid hydrophobicity is not critical for stabilization of hemoglobin. Polymerization properties of the variants in the deoxy form, however, were affected by the beta87 amino acid. Polymerization of Hb S beta87 Thr --> Leu and Hb S beta87 Thr --> Trp was preceded by a delay time like Hb S, while Hb S beta87 Thr --> Asp did not show a delay time. In addition, changes in time required for half polymer formation (T1/2) as a function of hemoglobin concentration for Hb S beta87 Thr --> Asp were similar to that for beta87 Thr --> Gln. Hb S beta87 Thr --> Leu polymerized at a lower hemoglobin concentration than Hb S while beta87 Thr --> Trp and Hb S beta87 Thr --> Asp required much higher hemoglobin concentrations for polymer formation. Critical concentration required for deoxy Hb S beta87 Thr --> Asp polymerization was 6- and 2.3-fold greater than that for Hb S beta85 Phe --> Glu and Hb S beta88 Leu --> Glu, respectively. These results suggest that even though beta87 Thr is not a direct interaction site for beta6 Val in deoxy Hb S polymers, it does play a critical role in formation of the hydrophobic acceptor pocket which then promotes protein-protein interactions facilitating formation of stable nuclei and polymers of deoxy Hb S. PMID- 9398335 TI - Metal-catalyzed oxidation and mutagenesis studies on the iron(II) binding site of 1-aminocyclopropane-1-carboxylate oxidase. AB - The final step in the biosynthesis of the plant signaling molecule ethylene is catalyzed by 1-aminocyclopropane-1-carboxylate (ACC) oxidase, a member of the non heme iron(II) dependent family of oxygenases and oxidases, which has a requirement for ascorbate as a co-substrate and carbon dioxide as an activator. ACC oxidase (tomato) has a particularly short half-life under catalytic conditions undergoing metal-catalyzed oxidative (MCO) fragmentation. Sequence comparisons of ACC oxidases with isopenicillin N synthase (IPNS) and members of the 2-oxoglutarate Fe(II) dependent dioxygenases show an aspartate and two of six ACC oxidase conserved histidine residues are completely conserved throughout this subfamily of Fe(II) dependent oxygenases/oxidases. Previous mutagenesis, spectroscopic, and crystallographic studies on IPNS indicate that the two completely conserved histidine and aspartate residues act as Fe(II) ligands. To investigate the role of the conserved aspartate and histidine residues in ACC oxidase (tomato fruit), they were substituted via site-directed mutagenesis. Modified ACC oxidases produced were H39Q, H56Q, H94Q, H177Q, H177D, H177E, D179E, D179N, H177D&D179E, H211Q, H234Q, H234D, and H234E. Among those histidine mutants replaced by glutamine, H39Q, H56Q, H94Q, and H211Q were catalytically active, indicating these histidines are not essential for catalysis. Mutant enzymes H177D, H177Q, D179N, H177D&D179E, H234Q, H234D, and H234E were catalytically inactive consistent with the assignment of H177, D179, and H234 as iron ligands. Replacement of H177 with glutamate or D179 with glutamate resulted in modified ACC oxidases which still effected the conversion of ACC to ethylene, albeit at a very low level of activity, which was stimulated by bicarbonate. The H177D (inactive), H177E (low activity), D179E (low activity), and H234Q (inactive) modified ACC oxidases all underwent MCO fragmentation, indicating that they can bind iron, dioxygen, ACC, and ascorbate. The results suggest that MCO cleavage results from active site-mediated reactions and imply that, while H177, D179, and H234 are all involved in metal ligation during catalysis, ligation to H234 is not required for fragmentation. It is possible that MCO fragmentation results from reaction of incorrectly folded or "primed" ACC oxidase. PMID- 9398336 TI - Mutagenesis of some positive and negative residues occurring in repeat triad residues in the ADP/ATP carrier from yeast. AB - In AAC2 from Saccharomyces cerevisiae, nine additional charged residues (six positive, three negative) were neutralized by mutagenesis following the previous mutation of six arginines. Oxidative phosphorylation (OxPhos) in cells and mitochondria, the expression level of AAC protein, and the various transport modes of AAC in the reconstituted system were measured. Mutations are: within the first helix at K38A which is exclusive for AAC; K48A, and R152A, part of a positive triad occurring in the matrix portion of each repeat; two matrix lysines, K179M and K182I, and the negative triad helix-terminating residues, E45G, D149S, D249S. Cellular ATP synthesis (OxPhos) is nearly completely inhibited in K48A, R152A, D149S, and D249S, but still amounts to 10% in K38A and between 30% and 90% in the gly+ mutants K179M, K179I + K182I, and E45G. Comparison of the AAC content measured by ELISA and the binding of [3H]CAT and [3H]BKA reveals discrepancies in K48A, D149S, and D249S mitochondria, which provide evidence that these mutations largely abolish inhibitor binding. Also these mitochondria have undetectable OxPhos. Differently in K38A, CAT and BKA binding are retained at high AAC levels but OxPhos is very low. This reveals a special functional role of K38, different from the more structural role of R152, K48, D149, and D249. Transport activity was measured with reconstituted AAC. The electroneutral ADP/ADP exchange of gly- mutants is largely or fully suppressed in K48A, D149S, and D249S. K38A and R152A are still active at 18% and 30% of wt. The other three exchange modes, ATP/ADP, ADP/ATP, and ATP/ATP, are nearly suppressed in all gly- mutants but remain high in gly+ mutants. ATP-linked modes are higher than the ADP/ADP mode in gly+ but lower in gly- mutants, resulting in an exchange mode inversion (EMI). In the competition for AAC2 transport capacity, the weak ATP exporting modes are suppressed by the much stronger unproductive ADP/ADP mode causing inhibition of OxPhos. Together with previous results all members of three charge triads are now mutagenized, revealing drastic functional rotatory asymmetries within the three repeat domains. In the intrahelical arginine triad the third (R294A), in the positive matrix triad the second (R152A), and in the helix-terminating negative triad the first (E45G) still show high activity. PMID- 9398337 TI - Site-directed mutagenesis of the active site glutamate in human matrilysin: investigation of its role in catalysis. AB - Glu-198 of human matrilysin is a conserved residue in the matrix metalloproteinases and is considered to play an important role in catalysis by acting as a general base catalyst toward the zinc-bound water molecule, on the basis of mechanistic proposals for other zinc proteases. In the present study, Glu-198 is mutated into Asp, Cys, Gln, and Ala, and the zinc binding properties, kinetic parameters, and pH dependence of each mutant are determined in order to examine the role of Glu-198 in catalysis. The mutations chosen either modify (C and D) or eliminate (A and Q) the general base properties of residue-198. All the mutants bind 2 mol of zinc per mol of enzyme, indicating that Glu-198 is not crucial to the binding of the catalytic zinc to the enzyme. The value of kcat/Km for the E198D mutant is only 4-fold lower than that of wild-type enzyme at the pH optimum of 7.5, while that for the E198C mutant is decreased by 160-fold. The E198Q and E198A enzymes containing the mutations that have eliminated the nucleophilic and acid/base properties of the residue are still active, having lower kcat/Km values of 590- and 1900-fold, respectively. The decrease in activity of all the mutants is essentially due to a decrease in kcat. The kcat/Km values of the mutants as a function of pH display broad bell-shaped curves that are similar to the wild-type enzyme. The acidic pKa value is not greatly affected by the change in the chemical properties of residue-198. The similarity in the pH profiles for the mutant and wild-type enzymes indicates that the ionization of Glu-198 is not responsible for the acidic pKa. Ionization of the zinc-bound water may be responsible for this pKa since the three His ligands and the scaffolding of the matrilysin catalytic zinc site are different from that observed in carboxypeptidase A and would predict a lower pKa for the metal-bound water. If the zinc-bound water is the nucleophile in the reaction, the role of Glu-198 in catalysis may be to stabilize the transition state or act as a general acid catalyst after the rate-determining step. PMID- 9398338 TI - In vivo footprinting using N-ethyl,N-nitrosourea: improved resolution of the DNA protein interactions in the human gamma-globin gene promoter region. AB - N-Ethyl, N-nitrosourea (ENU) was used as a probing agent in conjunction with a modified ligation-mediated polymerase chain reaction in a new in vivo footprinting procedure. In the present work, we examined the promoter region of the human gamma-globin gene under both uninduced and hemin-induced conditions in K562 cells. In the course of comparing this method with the standard dimethyl sulfate (DMS) in vivo method and previously reported results, we were able to verify our new method. However, discrepancies between these methods were observed at the stage selector element, -50 region, of gamma-globin promoter. Our in vivo footprinting result showed DNA-protein interaction at this region under the hemin induced condition which was not revealed by the conventional DMS in vivo footprinting method. This approach, using ENU-modified in vivo footprinting, is now being applied to clarify the mechanism of cytotoxic drug-induced fetal hemoglobin augmentation. PMID- 9398339 TI - Determination of the dissociation constant of phosvitin-anti-phosphoserine interaction by affinity capillary electrophoresis. AB - We used affinity capillary electrophoresis (ACE) to study the interaction of a monoclonal anti-phosphoserine antibody (mAb) to a homopolyvalent antigen (hpAg), phosvitin. A model system, which allows the measurement of the true dissociation constant (Kd) in Ag excess based on measurement of migration shifts of mAb-hpAg complexes at different Ag concentrations in solution, is presented for the study of the interactions between a mAb and an Ag that has identical determinants. The experimental value of Kd (22.4 x 10(-6) M) obtained by ACE is shown to be in close agreement with the value (17.8 x 10(-6) M) obtained by the conventional immunoassay based on indirect competition enzyme-linked immunosorbent assay (ELISA). Moreover, the Kds of mAb-hpAg complexes were measured and shown to be independent of the applied electrical field strength. Thus, under conditions where the total Ag concentration is in large excess over the total Ab concentration and when certain requirements are fulfilled, this method offers the advantage of dealing with the determination of Kd for unlabeled mAb and homopolymeric Ag molecules in free solution rather than at the liquid-solid interface. PMID- 9398340 TI - A colorimetric assay for phosphate to measure amplicon accumulation in polymerase chain reaction. AB - We describe a rapid colorimetric method for the detection of PCR products. Color generation is complete within 5 min of mixing reagents with a PCR. The method is simple and does not require affinity capture steps or special labeling to be carried out. The color development can be monitored by eye or a simple spectrophotometer can be used to read sample absorbance. We demonstrate the method by its use in the amplification refractory mutation system (ARMS) analysis [C. R. Newton, A. Graham, L. E. Heptinstall, S. J. Powell, C. Summers, N. Kalsheker, et al. (1989) Nucleic Acids Res. 17, 2503-2515] of cystic fibrosis [J. R. Riordan, J. M. Rommens, B. Kerem, N. Alon, R. Rozmahel, Z. Grzelczak, et al. (1989) Science 245, 1066-1073] and factor V [R. M. Bertina, B. P. C. Koeleman, T. Koster, F. R. Rosendaal, R. J. Dirven, H. de Ronde, P. A. van der Velden, and P. H. Reitsma (1994) Nature 369, 64-67] allelic variants. PMID- 9398341 TI - A large-scale in situ hybridization system using an equalized cDNA library. AB - We have developed a large-scale in situ hybridization system in which all the procedures are carried out on a 96-well format: digoxigenin-labeled probes were synthesized from PCR-amplified templates, sections were mounted on 96-well plates, and hybridization and immunohistochemistry protocols were carried out in each well of the plates. This system in combination with equalized (normalized) cDNA library as the source for the probes enables us to identify the cellular distribution of many mRNAs in various tissues rapidly and efficiently. Thus, this system may be a novel cloning method of identify genes differentially expressed in many tissues. In addition, this system has a potential to be automated. PMID- 9398343 TI - A set of constructed type spectra for the practical estimation of peptide secondary structure from circular dichroism. AB - While reliable methods have been developed for the estimation of globular protein secondary structure content from their far UV circular dichroism spectra, these are not suitable for the analysis of simple peptides. Model peptides have been measured in purely alpha-helical, beta-sheet, and coil form, and are often used for fitting the CD spectra of peptides, but these are mainly derived from homopolymers and exhibit side-chain-dependent characteristics that do not accurately represent the situation in natural sequences. We have attempted to reduce the side-chain bias inherent in these spectra by constructing a series of frequency-weighted "average" spectra from all available data on model peptides. These have proved quite satisfactory in estimating the secondary structure of a number of peptides. A computer program incorporating these spectra has been developed for practical use. PMID- 9398342 TI - Detection and analyses of ascorbyl radical in cerebrospinal fluid and serum of acute lymphoblastic leukemia. AB - We have detected and analyzed a free radical in human cerebrospinal fluid (CSF) of acute lymphoblastic leukemia (ALL) for the first time using electron paramagnetic resonance (EPR) at ambient temperature. We have also introduced an alternative capillary method to measure the radical. EPR spectra of the radical show a characteristic doublet with hyperfine coupling value of 1.8 G and g = 2.005. Based on EPR measurements, computer simulation, and literature values, we have determined that the species is ascorbyl radical (AsR). The radical has been investigated in CSF samples from ALL patients having no therapy, undergoing chemotherapy, and following therapy. Determination of the ascorbyl radical concentrations in CSF and serum was attempted using known concentrations of a nitroxyl radical. In addition, comparison in CSF and serum for ALL has been made along with statistical analyses of the data obtained. We found that AsR in CSF and serum has a strong correlation in patients undergoing chemotherapy (n = 57, r = 0.57, P < 0.0001). Ascorbate in CSF and serum show good correlation in patients having therapy but not for patients after therapy. PMID- 9398344 TI - Fluorometric assay of binding specificity of plant lectins to yeast cells by biotin-avidin system and its application to the classification of yeast cells. AB - A fluorometric assay of lectin binding to yeast cells is reported. The relative amount of biotinylated lectins bound to the yeast cells was estimated by enzyme activity using 4-methylumbelliferyl-beta-D-galactoside as a substrate for the lectin-bound beta-galactosidase through biotin-avidin interaction. Binding properties of 4 mannose-specific and 3 glucose/mannose-specific lectins to 22 different species of yeast cells were studied. The binding reaction of biotinylated lectins to the yeast cells was rapid and became constant within 10 min. Each lectin showed its characteristic binding specificity to each yeast species. The relative fluorescent intensities observed for 4-methylumbelliferone released by the action of bound beta-galactosidase were good indicators for the classification of yeast cells in quantitative base. We found that the yeast cells of the Saccharomyces genus can be classified into three groups, and those of Pichia were grouped into two groups. The present method can examine many samples simultaneously and be completed within 3 h. PMID- 9398345 TI - Reassessment of stereochemical configuration of natural phosphatidylglycerols by chiral-phase high-performance liquid chromatography and electrospray mass spectrometry. AB - Using chiral-phase high-performance liquid chromatography (HPLC) and electrospray ionization-mass spectrometry (ESI/MS), we have redetermined the stereochemical configuration of some natural and synthetic phosphatidylglycerols (PG). For this purpose, the synthetic and natural PG were converted to their bis-3,5 dinitrophenylurethanes (DNPU), which were separated by HPLC using two columns having chiral phases of opposite configuration, (R)-(+)- and (S)-(-)-1-(1 naphthyl)ethylamine polymers. The molecular species were identified by on-line negative-ion ESI/MS. Absolute configurations of the resolved peaks were assigned by comparison with the elution order of the corresponding 1(3)-monoacyl-sn glycerol enantiomers as bis-DNPU derivatives on the same column. The results clearly showed that the PG from cabbage leaf lipids and soybean phospholipids consisted of single R,S isomers (1,2-diacyl-sn-glycero-3-phospho-1'-sn glycerols), despite the presence of nonstereospecific phospholipase D in the tissues. On the other hand, the PG derived from egg yolk phosphatidylcholine and glycerol by transphosphatidylation with cabbage phospholipase D was a mixture of 45% R,S isomers (1, 2-diacyl-sn-glycero-3-phospho-1'-sn-glycerols) and 55% R,R isomers (1,2-diacyl-sn-glycero-3-phospho-3'-sn-glycerols). The PG from Escherichia coli lipids was a mixture of 89% R,S and 11% R,R isomers. The present study demonstrates that chiral-phase HPLC and negative-ion ESI/MS provide direct and unambiguous information about the configuration, identity, and quantity of molecular species in natural and synthetic PG. PMID- 9398346 TI - A colorimetric assay method for 3beta-hydroxy-delta5-steroid dehydrogenase. AB - 3beta-Hydroxy-Delta5-steroid dehydrogenase is an important enzyme of steroid hormone biosynthesis present in steroidogenic tissues like adrenal, testis, and ovary of vertebrates. The enzyme is assayed mainly by radiochemical substrates. Spectrophotometric assay is not adequately sensitive to detect the enzyme activity since it is often present in low levels. We have developed a simple colorimetric assay based on formazan formation due to the reduction of the tetrazolium salt. The reaction mixture containing the substrate, pregnenolone or dehydroepiandrosterone, NAD and iodonitrotetrazolium in 0.1 M Tris-HCl buffer (pH 7.8), and the enzyme extract is incubated for 1 h at 37 degrees C. Absorbance at 490 nm is read in a spectrophotometer. The enzyme activity was linear with time and protein concentration. The assay works well with adrenal tissue extract, whereas in the case of testis, Leydig cell preparation may be required. We have assayed the enzyme activity in the adrenal of rat, mouse, and gerbil. The method is two- to threefold more sensitive than the spectrophotometric assay. PMID- 9398347 TI - Interaction of immobilized avidin with an aequorin-biotin conjugate: an aequorin linked assay for biotin. AB - Biotinylated recombinant aequorin was used in the development of a heterogeneous bioluminescence binding assay for biotin. This assay is based on a competition between a biotinylated aequorin conjugate and biotin for the binding sites of avidin immobilized on solid particles. Dose-response curves were obtained that relate solid-phase aequorin activity to the concentration of biotin. Under certain experimental conditions these curves were biphasic; i.e., as the biotin concentration increased, the solid-phase aequorin activity first increased reaching a maximum and then decreased at higher biotin concentrations. This "hook" effect was observed with four different types of immobilization supports. The effect was more pronounced when low concentrations of aequorin-biotin conjugate were used, and diminished at a high conjugate concentration. This behavior indicates a possible positive cooperativity in the interaction between the immobilized avidin and biotin. Scatchard plot analysis was also consistent with a positive cooperativity mechanism. By using the ascending portion of the dose-response curve, the detection limit of the assay for biotin was 1 x 10(-15) M (100 zmol of biotin in the sample). PMID- 9398348 TI - Transfection of myoblasts in primary culture with isomeric cationic cholesterol derivatives. AB - Transfection of satellite cells from dog muscle (myoblasts) in primary culture has been optimized with respect to the position of the cholesteryl moiety along the polyamine chain of spermidine or spermine. Spermidine or spermine were derivatized with cholesterylchloroformate giving rise to three isomers in the case of spermidine and two isomers for spermine that were separated by reversed phase high-performance liquid chromatography (rp-HPLC). The position of the cholesteryl moiety was assigned by 13C-NMR and coelution with synthetic isomers of defined structure. The isomeric cationic lipids were evaluated for their transfection activity in myoblasts from dog muscle and a human lung epithelial cell line (A549) using plasmid DNA expressing the luciferase reporter gene. The results showed that the position of the cholesteryl moiety is of critical importance for efficient transfection of myoblasts in primary culture with isomers having a derivatized secondary amine being significantly more effective than those with a derivatized primary amine. On the contrary, differences in the A549 cell line were less pronounced and did not follow the same pattern. The results show that slight structural differences between cationic lipids lead to significantly different transfection efficiencies for myoblasts in primary culture. This may also represent an advantage in view of cell or organ targeting. PMID- 9398349 TI - Quantitation of the Escherichia coli methionine repressor-operator interaction by surface plasmon resonance is not affected by the presence of a dextran matrix. AB - The effect of a dextran matrix on the apparent rate constants measured for the interaction of the Escherichia coli methionine repressor, MetJ, with its immobilized consensus operator has been studied using surface plasmon resonance (SPR) in a commercial biosensor, BIACORE (Biacore AB). Based on the results of computer simulations, it has been proposed that such data can deviate from the expected simple kinetic behavior due to effects generated by the dextran matrix, used at the biosensor surface to anchor one of the interacting molecules. We have tested this possibility experimentally by measuring the apparent rate constants for the interaction of MetJ with its operator DNA on sensor chip surfaces with no dextran matrix or having matrices 30 or 100 nm thick. The data show that for the MetJ-operator interaction, the dextran matrix has no significant effect on the apparent rate constants measured and that comparative measurements using this technique are informative. PMID- 9398351 TI - Measurement of biochemical affinities with a Gill titration calorimeter. AB - A Gill titration calorimeter is evaluated as an instrument to determine in one experiment the equilibrium constant and the enthalpy change of a biochemical reaction. The dimensionless parameter kc (the product of the association equilibrium constant and the concentration of the reagent to be titrated; Wiseman et al., Anal. Biochem. 179, 131-137, 1989) is used to analyze the instrument performance. The analysis of simulated titration data corresponding to a simple model case shows that association equilibrium constants in the 10(2)-10(7) M-1 range may be determined when the kc parameter is between 1 and 1000. In addition we use a Monte Carlo approach to estimate the precision in the thermodynamic parameters of the reaction under study. The relative precision in the calculated constants ranges from 3 to 80% depending on the macromolecule concentration and kc value in the experiment. These results were checked with the study of the reactions of beta-trypsin with its inhibitor and ribonuclease A with cytidine 2' monophosphate and cytidine 3'-monophosphate. PMID- 9398350 TI - Site-specific fluorescence labeling of the beta2 adrenergic receptor amino terminus. AB - A modified human beta2 receptor, designated 0K-beta2, was developed for site specific labeling at the amino terminus with amine reactive fluorescent probes. 0K-beta2 has the following modifications: (1) all 16 lysines in the wild-type beta2 receptor were mutated to arginines, (2) a FLAG epitope preceded by a cleaved hemagglutinin signal sequence was fused to the amino terminus, and (3) a hexahistidine tail was added to the carboxyl terminus. The FLAG epitope and hexahistidine tail were added to facilitate purification while lysine to arginine mutations eliminate potential labeling sites for amine-reactive fluorescent probes. The remaining primary amines in the 0K-beta2 receptor, the amino terminal amine and the epsilon-amine of Lys3, both reside in the amino-terminal FLAG epitope. The 0K-beta2 receptor expressed in Sf9 insect cells exhibited ligand binding and G-protein coupling characteristics similar to the wild-type beta2 receptor. The modified receptor was labeled with fluorescamine, an amine-reactive fluorescent probe. Proteolysis with factor Xa showed that labeling was confined to the amino terminus of the 0K-beta2 receptor. Our results demonstrate site specific fluorescamine labeling at the amino terminus of the 0K-beta2 receptor, a lysine-depleted beta2 receptor that retains functional characteristics of the wild-type receptor. PMID- 9398352 TI - Radiolabeling of the lipids of chinese hamster ovary cells with the probe [3 (trifluoromethyl)-3-(m-[125I]iodophenyl)diazirine]. AB - [125I]TID [3-(trifluoromethyl)-3-(m-[125I]iodophenyl)diazirine] is a commercially available, hydrophobic, photoactivatable, gamma-emitting reagent mostly used to label protein hydrophobic domains. It has also been used to radiolabel the phospholipids of lung surfactant (Gilliard et al., Anal. Biochem. 193, 310-315, 1991). Since a nonspecific, highly sensitive, lipid-labeling probe would be a very useful tool to investigate lipid-protein interactions in biological membranes, we characterized further the [125I]TID-labeling products of lipids from cultured Chinese hamster ovary cells (IR-CHO). After labeling of whole cells, TLC analysis followed by autoradiography enabled detection of sphingomyelin, phosphatidylcholine, phosphatidylinositol, phosphatidylserine, phosphatidylethanolamine, cardiolipin, diglycerides, cholesterol and its esters, and triglycerides. Analysis of the radioactivity associated with the saponification products of different lipids showed that [125I]TID was mostly (80%) extracted with the fatty acid moiety of the lipids whereas 20% remained associated with the hydrosoluble moiety. Similar radioactivity profiles were observed after labeling of whole cells or extracted and liposome-reconstituted lipids; the [125I]TID probe was able to diffuse in all intracellular organelles. Labeling was not equivalent between the different lipid classes, and it appeared that the amount of associated radioactivity correlated well with the degree of lipid unsaturation. This was confirmed by studying [125I]TID incorporation in phosphatidylcholines of different chain length and unsaturation. Taken together, our data demonstrate that [125I]TID can be used as a radiolabel for lipids in cultured cells. It is rapidly incorporated in the hydrophobic part of membranes, diffuses into all cellular compartments, and labels all lipid classes, including phospholipids, cholesterol, and glycerides, with a sensitivity in the nanomolar range. PMID- 9398353 TI - Optimized alcoholytic deacetylation of N-acetyl-blocked polypeptides for subsequent Edman degradation. AB - N-terminal protein acetylation is a common posttranslational modification, blocking Edman degradation during sequencer analysis. Use of mass spectrometry allows the analysis also of acetyl-blocked polypeptides; however, for large proteins mass spectrometry is not always informative, and deacetylation by chemical pretreatments is desirable for making direct sequencer analysis possible. For this purpose, alcoholytic deacetylation is attractive. In the present work, we have studied the optimal conditions for specific removal of the acetyl group without extensive cleavage of peptide bonds in general. We find that incubation with trifluoroacetic acid in methanol (1:1, by volume) at an elevated temperature ( approximately 47 degrees C) for 2-3 days results in efficient deacetylation allowing direct application to sequencer analysis with initial yields up to approximately 50% of the amount applied for deblocking. Deacetylation compared to internal peptide bond cleavage is often high, as evaluated by recoveries of residues from the deblocked sequence over those from the background, and this applies to both peptides (up to the order of 10:1 for the specific residue versus the background) and proteins (>2:1). Although yields may still vary and some sequences be only partly susceptible to the chemistry, this deblocking can in many cases allow unambiguous interpretation of N terminally acetyl-blocked sequences. PMID- 9398354 TI - Fluorescence detection of calcium-binding proteins with quinoline Ca-indicator quin2. AB - We have established a fluorescence method to detect calcium-binding proteins by making use of the quinoline Ca indicator quin2. Authentic calcium-binding proteins were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and then electrophoretically transferred onto polyvinylidene difluoride membranes. Transfers were incubated with nonradioactive calcium ions, then with quin2 to detect the calcium-binding proteins as fluorescent bands by illumination with UV light. Calmodulin and parvalbumin of EF hand conformation calcium-binding type were clearly identified. Quin2 distinguished smooth muscle alpha-actinin from skeletal muscle alpha-actinin; the former was faintly fluorescent, having a low affinity for calcium ions. In whole myofibril preparations from skeletal muscles, troponin-C, connectin (titin), and nebulin were intensely fluorescent, being shown to have calcium-binding ability. The fluorescence method is an accurate, safe, and simple procedure to detect the binding of calcium ions to proteins following electrophoresis. The overlay technique described can be completed within 15 h and detects as little as 38 ng/well of troponin-C in the starting sample. PMID- 9398355 TI - Macroscopic orientation of natural and model membranes for structural studies. AB - One approach for obtaining high-resolution structural and functional information for biomembranes and their proteins is by static solid-state NMR of oriented systems. Here, a general procedure to align fully functional biological membranes containing large membrane proteins (Mr >30,000) is described. The method, based on the isopotential spin-dry ultracentrifugation technique, relies on the centrifugation of membrane fragments onto a support with simultaneous, or subsequent, partial evaporation of the solvent which aids alignment. The quality of orientation, as shown by the mosaic spread of the samples, was monitored by static solid-state 31P NMR for the phospholipids and by 2H NMR for a deuterated retinal in bovine rhodopsin. The generality of this method is demonstrated with three different membranes containing bovine rhodopsin in reconstituted bilayers, natural membranes with the red cell anion exchange transport protein in erythrocytes, band 3, and the nicotinic acetylcholine receptor. PMID- 9398356 TI - A packaging system for SV40 vectors without viral coding sequences. AB - SV40 vectors have been used as expression vectors for mammalian cells since the early 1980s. More recently, they have been used as gene transfer vectors in mice and in human peripheral blood cells. Here we described a system for packaging SV40 vectors without viral coding sequences. Recombinant adenovirus-expressing SV40 capsids can effectively package plasmids that contain the SV40 replication origin. The final yield of infectious SV40 vector is about 3 x 10(5), with a SV40 to adenoviral vector ratio of about 1000:1. Helper adenoviruses can be effectively heat-inactivated with no effect on the infectivity of SV40 vectors. PMID- 9398357 TI - Continuous assay of proteases using a microtiter plate fluorescence reader. PMID- 9398358 TI - A modified pGEX vector with a C-terminal histidine tag: recombinant double-tagged protein obtained in greater yield and purity. PMID- 9398359 TI - A set of pBR322-compatible plasmids allowing the testing of chaperone-assisted folding of proteins overexpressed in Escherichia coli. PMID- 9398360 TI - Assay of protein phosphatase activities with phosphopeptide-magnetic particle conjugates. PMID- 9398361 TI - Feeding chases and food allocation in Adelie penguins, Pygoscelis adeliae AB - Among penguins, well-developed feeding chases where chicks run after adults and beg for food are found only in the genera PygoscelisAptenodytesFeeding chases involving adult Adelie penguins, with two chicks, compared to feeding chases involving those with one chick, were longer, occurred further from the breeding group and included more runs between each feed. Feeding chases were more likely when chicks were close together, and which chick was fed was more likely to switch after a feeding chase. These results suggest that feeding chases serve to minimize harassment from other chicks during feeding and to reduce the effects of sibling competition.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9398362 TI - A field test of threat sensitivity in a marine gastropod AB - In the Mingan Islands, the whelk Buccinum undatum displays defensive manoeuvres to both contact and water-borne chemical cues from the predatory asteroid Leptasterias polarisIn spite of this, whelks occasionally aggregate in great numbers near L. polaris while it is ingesting a prey; they then attempt to steal food from their predator and also wait for leftovers. In this study, the response of whelks in different types of encounters with L. polaris was examined to test the hypothesis that whelks are sensitive to the magnitude of the threat their predator represents. In a field experiment, whelks consistently fled both non feeding and feeding L. polaris (asteroids used were consuming small prey items that were unlikely to provide food for whelks). When current flow was stable, whelks fled more directly down current and more frequently displayed violent defensive behaviours, in response to non-feeding L. polariswhich presented a higher risk, than in response to feeding asteroids (lower risk; 47% versus 2%). Consequently, whelks tested with non-feeding asteroids more rapidly distanced themselves from the predators than did whelks tested with feeding asteroids. In a field survey, there were more active whelks in the vicinity of cruising (higher risk) than stationary (lower risk) L. polaris (53% versus 14%). Among those whelks that were active, defensive behaviour patterns such as shell rocking and leaping escape movements were frequently shown by whelks near cruising predators (69%), but never by whelks near stationary predators (0%). The discriminative capabilities apparent in these results are likely to be adaptive, because they enable whelks to limit the cost of escape responses while still keeping predation risk low, and also because they facilitate a close association with L. polaris from which the whelks receive feeding benefits.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9398363 TI - Does brood reduction provide nestling survivors with a food bonus? AB - Siblicide is common in many asynchronously hatching birds, including brown pelicans, Pelecanus occidentalisMost adaptive models of siblicide, and of brood reduction in general, tacitly assume that parental deliveries of food remain fixed, and therefore that supplies to seniors remain unchanged or increase after a junior's death. If parents match deliveries to brood size, however, then seniors may get less food following brood reduction. To test the assumption that parental deliveries remain constant, and to determine how brood reduction affected seniors, food deliveries to control (three-chick) and experimentally reduced (two-chick) broods of brown pelicans were compared. Parents brought less food to reduced than to control broods. Similarly, a literature review revealed that avian parents generally delivered less food to smaller than to larger broods of experimentally altered sizes. In brown pelicans, when food deliveries decreased following brood reduction, second-hatched ('B') chicks received less food, but first-hatched ('A') chicks' supplies remained unchanged. B-chicks may have gained more in control broods by appropriating the extra food that last hatched ('C') chicks otherwise would have gained. If brood reduction decreases fighting, nestlings may gain more net energy even if they receive less food. Fighting did not decrease significantly in reduced broods, perhaps because fledging averaged one chick/brood, so A and B-chicks still competed intensely. Alternatively, small sample sizes may have prevented detection of differences in fighting. Seniors did not concentrate their attacks on C-chicks, perhaps because seniors may benefit by delaying the decreases in parental food deliveries that follow brood reduction.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9398364 TI - Responses to conspecific and heterospecific olfactory cues in the swordtail Xiphophorus cortezi AB - Female Xiphophorus cortezi responded to olfactory cues from both conspecific males and heterospecific X. nigrensisX. montezumae males when given a choice between the stimulus and water. When given a choice between the conspecific and heterospecific cues, however, females demonstrated a strong preference for the conspecific stimulus. Of the two heterospecific species, females responded more strongly to their close relative X. nigrensis than they did to the more distantly related X. montezumaeMate recognition in northern swordtails is evidently not a simple process based upon a response to one variable, but the outcome of complex responses to information from at least both visual and olfactory cues.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9398365 TI - Social mating system affects the frequency of extra-pair paternity in house wrens AB - We tested two hypotheses regarding the effect of the social mating system on extra-pair paternity in an Illinois population of house wrens, Troglodytes aedonAccording to the genetic-quality hypothesis, polygynous males are of higher quality than monogamists, and monogamously paired females, in an attempt to obtain high-quality genes, should have a greater proportion of extra-pair nestlings in their nests than polygynously paired females. According to the trade off hypothesis, polygynists, with temporally overlapping nests, will have a greater proportion of extra-pair nestlings in their nests than monogamists, because polygynists have difficulty guarding one or both of their social mates. DNA fingerprinting revealed that extra-pair paternity was most frequent in secondary nests of polygynists. The proportion of secondary broods with extra pair nestlings increased with the temporal overlap of polygynists' nests, although this trend was not significant. Both results are consistent with the trade-off hypothesis but not with the genetic-quality hypothesis. We did not address the effects of genetic quality on male success at siring nestlings in the nests of other males. Although the trade-off hypothesis focuses on male mate guarding, female behaviours may also affect frequencies of extra-pair paternity. Secondary females may compensate for reduced male defence by engaging in extra pair copulations with neighbours to reduce the likelihood that neighbours destroy their nests. Thus, in house wrens, female participation in extra-pair copulations in combination with male mate-guarding constraints may generate higher levels of extra-pair paternity in secondary broods than in primary polygynous or monogamous broods.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9398366 TI - Parent-offspring recognition in tree swallows, Tachycineta bicolor AB - Parent-offspring recognition appears to be highly developed in species in which the risk of misdirecting care is high (e.g. colonial species). Some of the best evidence for this relationship comes from comparative work on swallows of the family Hirundinidae. Using methods followed in earlier studies, we determined whether parent-offspring recognition occurs in the tree swallow, Tachycineta bicolora non-colonial species closely related to the highly colonial bank swallow, Riparia ripariaand the solitary rough-winged swallow, Stelgidopteryx ruficollisParents did not discriminate between playbacks of the calls of their own versus non-related nestlings. However, older nestlings called more in response to playback of parental calls than non-parental calls, suggesting that they recognized their own parents. Despite significant individual variation in parental and nestling calls, variation in tree swallow nestling calls was lower than analogous calls in the bank swallow. Our results provide further support for a positive relationship between recognition, individual variation in call structure and coloniality.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9398367 TI - Hormonal dynamics during mate choice in the northern pintail: a test of the 'challenge' hypothesis AB - In previous mate choice experiments, we found no relationship between dominance rank and pairing success in male northern pintails, Anas acutaOnce chosen by a female, however, males became aggressive, initiated fights with higher-ranked males and quickly established dominance. In the present study, we tested a variation of the 'challenge' hypothesis, that the behavioural stimuli associated with acquiring and defending a mate induce an increase in testosterone level, which in turn facilitates aggressive behaviours required for males to establish dominance. We measured plasma hormone levels (testosterone, dihydrotestosterone, luteinizing hormone and corticosterone) before and after mate choice in two experiments in which males competed for a single female (experiments 1 and 2) and in a control experiment in which no female was introduced (experiment 3). We used groups of either three adult males (experiment 1) or one adult and two yearling males (experiments 2 and 3). Contrary to expectation, in experiment 1, plasma levels of corticosterone increased significantly and testosterone levels decreased in chosen males following mate choice. The magnitude of change in corticosterone was positively correlated with the rate of aggression by males. Chosen adult males in experiment 2 showed similar patterns of hormone change (corticosterone increase and testosterone decrease), although not all changes were significant. Hormone levels of unchosen yearlings in experiment 2 and control adults and yearlings in experiment 3 showed no changes. The results are consistent with the hypothesis that behavioural stimuli associated with successful pair formation induce a transitory increase in circulating levels of corticosterone, which in turn mediates the behavioural response of increased aggression leading to the establishment of dominance following mate choice. A short-term increase in corticosterone may be adaptive in this situation because it would mobilize energy stores needed by the male to defend the new pair bond and establish dominance.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9398368 TI - Signalling displays during predator-prey interactions in a Puerto Rican anole, Anolis cristatellus AB - We examined conspicuous signalling displays in the context of predator-prey interactions. To determine in which context Puerto Rican crested anoles, Anolis cristatellusperform conspicuous signals, we exposed wild lizards to a model of a natural snake predator. The lizards gave six behavioural responses to the model: immobility, predator inspection, flight, lateral face-off, dewlapping and push ups. They displayed significantly more push-ups and push-up bouts in the presence of the snake model. Alternative theories regarding the function of conspicuous signals in A. cristatellusthe flash concealment and predator deterrent hypotheses, were also tested. The flash concealment hypothesis proposes that the sudden display exhibition of signalling behaviour followed by the flight of the animal may confuse the predator about the position of the prey, thus causing the predator to abort the attack. The pursuit deterrent hypothesis contends that because the chances of the predator successfully attacking its prey decrease when the prey is aware of the incoming predator, prey have evolved signalling behaviours that communicate to the predator that it has been detected, therefore discouraging the attack. Results supported the use of push-ups, dewlapping, lateral face-off and predator inspection as predator deterrent signals. During the recognition phase of a predatory encounter, A. cristatellus may rely more on behavioural signals than on flight to avoid predation. Because the predator deterrent signals are the same as the signals used in social interactions, it is suggested that predation pressure may have reinforced the effects of sexual selection in the evolution of Anolis signalling displays.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9398369 TI - Scent-marking by coyotes, Canis latrans: the influence of social and ecological factors AB - We observed 49 coyotes, Canis latransfrom five resident packs for 2456 h and five transient coyotes for 51 h from January 1991 to June 1993 in the Lamar River Valley, Yellowstone National Park, Wyoming, U.S.A. During these observations we recorded 3042 urinations, 451 defecations, 446 ground scratches and 743 double marks. The rate of scent-marking (via urination) was influenced by the social organization (resident versus transient) to which the coyote belonged, the social class (alpha, beta or pup) of the animal and the time of the year. Transient coyotes scent-marked at a lower rate than did members of a resident pack. Within the resident packs, alpha coyotes scent-marked at a higher rate than beta coyotes (adults and yearlings subordinant to alphas, but dominant over pups) and pups. Alpha coyotes increased their rate of marking during the breeding season; beta and pup coyotes performed scent-marks at a relatively constant rate throughout the year. There was no influence of social class or time of year on the rate of defecation. The rate of double-marking was highest among alpha coyotes with a peak during the breeding season. Alpha coyotes ground-scratched at a higher rate than did beta and pup coyotes. Alpha and beta coyotes scent-marked more than expected along the periphery of the territory compared to the interior; pups marked in the interior and edge in proportion to expected frequencies. Double marking and ground-scratching were higher than expected along the periphery of the territory. The distribution of defecations was not different from expected along the edge versus the interior of the territory. Pack size did not influence the rate of scent-marking performed by individuals in the pack or by the alpha pair. We concluded that alpha coyotes were the primary members of the resident pack involved in scent-marking. The large coyote packs and the high rate of marking by the alpha pairs were parallel to the scent-marking behaviour displayed by wolves, C. lupusto a greater extent than previously reported. Scent-marks appear to provide internal information to the members of the resident pack (internal map of territory, breeding condition, reproductive synchrony) and enhance demarcation of territorial boundaries.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9398370 TI - Species-specific footdrumming in kangaroo rats: Dipodomys ingens, D. deserti, D. spectabilis AB - Footdrumming was compared in three allopatric species of kangaroo rat, Dipodomysfrom three habitats. Analysis of footdrumming recordings revealed species-specific patterns of drumming ranging from single thumps to individual footdrumming signatures. The desert kangaroo rat, D. desertidrums single thumps spaced 0.25-0.30 s apart that are sometimes introduced with a short footroll. The giant kangaroo rat, D. ingensdrums long footrolls that can average over 100 drums at 18 drums/s. The banner-tailed kangaroo rat, D. spectabilisdrums three to 38 footdrums in a footroll combined into sequences of two to 12 footrolls at a rate of 17 drums/s. In playback tests, all three species stood in alert postures and entered the burrow in response to footdrumming of their own and the other species. The rats also responded in species-specific ways. Dipodomys spectabilisdrummed to its own species' footdrumming, but not to playbacks of the single drums of D. desertiInstead of footdrumming to playbacks of its own species, D. deserti approached the speaker more frequently than did either of the other two species. Dipodomys ingens footdrummed equally to all footdrumming playbacks. The species' differences reflect differences in social tolerance and spacing. Dipodomys deserti rarely engages in footdrumming exchanges and chases visitors from the burrow. Dipodomys spectabilis engages in frequent footdrumming exchanges and some chases, and D. ingens seems to tolerate close neighbours and footdrums periodically.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9398371 TI - Testosterone implants increase song but not aggression in male Lapland longspurs AB - Lapland longspurs, Calcarius lapponicusare tundra-nesting arctic birds with an extremely short breeding season. Males show a pronounced brief peak of plasma testosterone early in the season. We studied the effect of exogenous testosterone on song and aggression in free-living male Lapland longspurs. We gave high testosterone implants to 16 birds, low-testosterone implants to seven birds, and empty or no implants to 33 birds. The implants resulted in significantly different plasma testosterone levels during incubation (high-testosterone mean=20.79±3.4 ng/ml, low-testosterone mean=2.54±0.5 ng/ml, control mean=0.53±0.1 ng/ml). High-testosterone birds had larger cloacal protuberances than controls, but low-testosterone birds did not. We measured aggression and song with simulated territorial intrusions during incubation, a mean of 12.03±1.45 days after the insertion of implants. High-testosterone birds and low-testosterone birds were no more aggressive than controls. However, high-testosterone birds, but not low-testosterone birds, were significantly more likely than controls to sing. These results suggest that the high, brief testosterone peak of Lapland longspurs may be related more to song than to territorial aggression. Alternatively, both aggression and song may be testosterone-influenced, but aggression may be more readily suppressed during incubation. The hormone-behaviour patterns demonstrated in this study may be adaptations for breeding in the short arctic summer.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9398372 TI - Non-nutritional maternal support in the brown long-eared bat AB - Adult female brown long-eared bats, Plecotus aurituswere taken into captivity over a 3-year period and housed in two free-flight enclosures. They were maintained in small groups, each roosting in a single, heated roost box while monitored by an infra-red sensitive video camera. The predicted percentage of records spent by lactating females in direct contact with the young on day 1 of lactation did not differ significantly from 100%. This declined to 13% on day 50 of lactation. Over time, the mothers groomed the young less. Lactating females visited the roost more times per night, but spent less time self grooming than non-reproductive females. The total amount of grooming behaviour (estimated as the percentage of records spent in self grooming plus those allocated to grooming of the young) for lactating females was 50% of the value for non-reproductive females. In general, care-giving behaviours declined with the progress of lactation. The temporal expression of these behaviours was opposite in direction to that of the expected energetic demands of milk production.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9398373 TI - The role of bright plumage in male-male interactions in the ring-necked pheasant AB - The brightness and colour of birds' plumages have been considered sexually selected traits, indicating health, condition or parasite resistance. However, recent studies with pheasants, Phasianus colchicussuggest they are not signals used by females in mate choice. Instead, males might rely on plumage when assessing the quality of competitors. In this study, bright and experimentally dulled males were presented to a group of captive male pheasants to determine the response to differences in plumage brightness of the intruder. Males in the group directed more aggression to the experimentally dulled males than they did to any other males. This may be partly because they considered the dull males as novel males. When the bright and the dull males were both unknown, both still received more aggression than the average for any individual in the group, but dull males were attacked by more males. Bright males were attacked more by the dominants and dull males by the subordinates. The results show that plumage brightness may affect individual recognition, but also that it is used by males to assess the quality of competitors. Male-male interactions, therefore, may have played a role in the evolution of plumage brightness, either in the context of competition for mates or for resources when males gather into unisexual groups.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9398374 TI - Capuchin monkeys, Cebus apella fail to understand a cooperative task AB - We investigated whether capuchin monkeys cooperate to solve a task and to what extent they take into account the behaviour of another individual when cooperating. Two groups of capuchin monkeys (N=5 and 6) were tested in a task whose solution required simultaneous pulling of two handles which were too far from one another to be pulled by one monkey. Before carrying out the cooperation study, individual monkeys were trained to pull one handle (training phase 1) and to pull two handles simultaneously (training phase 2) for a food reward. Nine subjects were successful in training phase 1, and five in training phase 2. In the cooperation study seven subjects were successful, that is, pulled one handle while a companion pulled the other. Further analyses revealed that capuchins did not increase their pulling actions when a partner was close to or at the other handle, that is, when cooperation might occur. These data suggest that capuchin monkeys acted together at the task and got the reward without understanding the role of the partner and without taking its behaviour into consideration. Social tolerance, as well as their tendency to explore and their manual dexterity, were the major factors accounting for the capuchins' success.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9398375 TI - Great spotted cuckoos improve their reproductive success by damaging magpie host eggs AB - Adult great spotted cuckoos, Clamator glandariusdamage the eggs of their magpie, Pica picahost without removing or eating them. The number of damaged magpie eggs was recorded in 360 parasitized nests of which 62.2% contained between one and eight damaged magpie eggs. Egg-destroying behaviour may be adaptive if it reduces nestling competition and/or enhances the hatching success of the cuckoo. To clarify the role of egg destruction for the reproductive success of great spotted cuckoos, unparasitized magpie nests were experimentally parasitized (without egg damage) by introducing cuckoo eggs or chicks. Egg damage was common in parasitized nests but the eggs were not damaged by the hosts. Egg damage increased the breeding success of the cuckoos, by both reducing the number of competing host chicks in the nest and increasing the likelihood that late-laid cuckoo eggs would hatch.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9398376 TI - Cohort size and the allocation of social effort by female mountain baboons AB - Dunbar (1992,Behav. Ecol. Sociobiol.33, 35-49) argued that constraints on social time limited the size to which savannah baboon, Papio cynocephalustroops in any given population could grow before fissioning. Since this should be reflected in population structure, we have elsewhere (Henzi et al. 1997a, Anim. Behav. 53, 525 535) constructed a model, based on a rising probability of fission, that fits the observed distribution of troop sizes of mountain baboons, P. c. ursinusin the Drakensberg mountains of South Africa and which predicts that the probability of fission will rapidly increase once a troop has more than 23 members (or 8.7 females). We test this prediction in this paper. Since Dunbar argued that females will drive fission once they cannot engage in the grooming necessary to sustain alliances, we compared the grooming interactions of adult females from four troops in the Drakensberg mountains. The mean female grooming clique size reached an asymptote at 7.4 females, so that females in cohorts of eight or more no longer attempted to groom all other females, and mean grooming bout length declined as the cohort grew to 7.9 females and then increased again. These values are coincident with the female cohort size predicted by our model of troop growth and fission. We argue that females attempt to groom all other females as well as sustain closer relationships with a few females through longer bouts of reciprocated grooming. When the demands of grooming all other females reduce bout length to a point when no reciprocated bouts are possible, female clique size is capped. As a troop continues to grow, the mechanical difficulties involved in gaining access to grooming partners leads to a reduction in the diversity of grooming relationships. This weakening of the total female network, as cliques become more differentiated, is likely to facilitate fission. We conclude that our data provide the first within-population validation of Dunbar's hypothesis concerning the mechanism underpinning fission. In the Drakensberg, where there is no advantage to female coalitions, we propose, as an amendment, that females will leave a troop not to escape local competition, but to follow a male with whom they have a close friendship.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9398377 TI - Motivational aspects of individual variation in response to nestboxes by laying hens AB - Laying hens, Gallus gallus domesticusshow individual variation in pre-laying behaviour including their ultimate choice of nest site. In housing systems with nestboxes, the majority of hens make a small number of long visits to nestboxes and lay their eggs therein, but some hens make many short visits and occasionally lay outside the nestbox. We investigated the motivational basis of this individual variation using six consistent hens which always laid in nestboxes and six inconsistent hens which sometimes laid outside nestboxes. Each hen was housed in a pen (containing either no nestbox, a semi-enclosed nestbox or an enclosed nestbox) with access to a ring-shaped tunnel which increased the opportunity to perform locomotor activity. Access to the tunnel could be restricted by narrowing the doorway to 140, 125, 110 or 95 mm (compared with a mean hen width of 114 mm). In trials with no nestbox, there was no difference in the pre-laying behaviour of consistent and inconsistent hens. Narrowing the doorway reduced the number of visits to the tunnel, but all hens persisted in visiting the tunnel and doorwidth had no effect on time spent therein. With both designs of nestbox, however, inconsistent hens visited the tunnel more often than consistent hens prior to oviposition, and continued to pass the narrowest doors to enter the tunnel, whilst consistent hens would not pass doors of 110 or 95 mm. After oviposition, there was no difference in the two groups' behaviour in any treatment and no hens would pass doors of either 110 or 95 mm to visit the tunnel. Individual variation in nest-site choice, therefore, appeared to result from different perception of nestboxes rather than lower nesting motivation. Inconsistent hens worked as hard as consistent hens to perform pre-laying locomotion, but appeared to be less responsive to the cues provided by nestboxes than consistent hens, because they persisted with pre-laying locomotion when provided with either design of nestbox.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9398378 TI - Mole-rat harderian gland secretions inhibit aggression AB - The harderian gland secretions of mole-rats, Spalax ehrenbergiusually released by self-grooming, include odorous substances which are sex dependent. Male secretions were the most attractive to both sexes, while female secretions were attractive to males but not to other females. The rate of attacks by females towards intact males was higher than towards males whose harderian gland had been removed. However, grooming by intact male mole-rats decreased the rate of attacks by their opponents, while grooming by males without harderian glands did not; thus the male harderian secretions appear to have appeasement qualities. Grooming by females with and without harderian glands failed to reduce aggression. Unlike intact males, those without harderian glands had almost no volatiles on their fur, and thus are probably not considered to be a threat to conspecifics. Gas chromatography spectra showed that substances of harderian origin were added to the fur during grooming. Some of these substances remained on the fur long after the animal ceased grooming, and appear to give the animal its specific odour, but some volatile substances peaked briefly after grooming, and were probably responsible for the decline of aggression that occurred after grooming. Although grooming has long been considered to be a displacement activity, we suggest that in the mole-rat its performance is too risky to be merely this, and it has acquired the meaning of appeasement through the release of chemical cues.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9398379 TI - Territorial intrusions and copulation behaviour in the great skua, Catharacta skua AB - Birds of many species intrude on to territories in order to obtain extra-pair copulations, and frequent within-pair copulations are thought to be a response to cuckoldry where mate guarding is not possible. Great skuas are colonial birds in which females are left alone in the breeding territory while males forage for the pair. Opportunities for cuckoldry are therefore numerous, and it could be predicted that sperm competition should be intense in this species. We tested several hypotheses that attempt to explain territorial intrusions by female great skuas. Few intrusions coincided with the main fertile period and extra-pair copulations were almost never solicited, and therefore the sperm competition hypothesis was rejected. Only 0.9% of the copulations observed (N=339) were extra pair. Thus opportunity for cuckoldry is a very poor predictor of the intensity of sperm competition, in spite of the relevance given to this factor in the literature. Of three extra-pair copulations observed, two involved unpaired territorial males. This suggests that genetic benefits were not the aim of unfaithful females. All three were preceded by courtship feeding, while only 26% of within-pair copulations followed successful food begging. This statistically significant difference constitutes evidence for a trade of copulations for food in a monogamous bird. Evidence is presented supporting hypotheses that females intrude on to territories in order to induce males to give away some food, and to become familiarized with potential partners for future breeding seasons.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9398380 TI - Laterality in detour behaviour: interspecific variation in poeciliid fish AB - We measured whether males of five species of poeciliid fish made detours to the right or left of a vertical-bar obstacle in order to approach a group of females. Three of these species, Gambusia holbrookiGambusia nicaraguensis and Poecilia reticulata showed a significant bias to the left, whereas Brachyrhaphis roseni and Girardinus falcatus showed a significant bias to the right. When tested for direction of turning in front of an opaque barrier, or when a dummy predator was used as a target in a detour test, G. holbrooki and G. falcatus showed similar biases to the right (opaque barrier) and left (predator), thus suggesting that the difference observed when females were used as a target could arise from species differences in the degree of sexual motivation in a novel environment. The two species that showed bias to the right with the females were less likely to exhibit sexual behaviour when placed in a novel environment. Moreover, manipulation of the factors affecting the relative strength of sexual motivation and of fear of a novel environment, such as how long fish were maintained in captivity or in the test apparatus before being tested, caused shifts in the direction of the lateral asymmetries. These results suggest that the presence of functional asymmetries in behaviour could be widespread among vertebrates and that the direction of such asymmetries tends to be strikingly similar in closely related species, thus supporting the hypothesis of an early evolution of laterality in brain and behaviour.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9398381 TI - Behavioural responses of Eurasian treecreepers, Certhia familiaris, to competition with ants AB - Competition for a specific resource that is essential for the survival of both the competitors may be intense even between very dissimilar taxa. However, the importance of the effects caused by such interspecific competition has seldom been emphasized. These effects can appear as differences in individual foraging behaviour during the breeding season, which can result in critical variation in fitness. In this study we examined the effects of wood ants (Formica rufa group) on the abundance of other invertebrates on tree trunks and on the foraging site selection of breeding Eurasian treecreepers, which use the same habitat as wood ants. Arthropods were scarcer on the trunks with ants present; the treecreepers avoided these trunks and foraged for a shorter time on trunks with ants than on trunks without ants. We also tested experimentally the existence of competition between ants and treecreepers by comparing the foraging behaviour of breeding treecreepers on spruce trunks with ants, without ants and with experimentally reduced numbers of ants. On average arthropods were scarcest on trunks with ants present. Male treecreepers also foraged for a shorter time on spruce trunks with ants. The reduction in ant numbers allowed food resources on trunks to recover over a week and led to longer foraging times of the treecreepers on these trunks than on trunks with ants present. The longest treecreeper visits were on trunks without ants. Our results suggest that competition between two very different taxa may be effective in determining the behaviour of foraging individuals.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9398382 TI - Male aerial display and reversed sexual size dimorphism in the dunlin AB - Reversed sexual size dimorphism, with males smaller than females, is common in waders. The aerial display hypothesis suggests that sexual selection in males favours aerial agility, and hence small size, in species with male display flights. We tested this hypothesis in the dunlin, Calidris alpinaDisplay flights were uncommon in the early breeding season but increased markedly when females began laying. Male display areas were largely overlapping, and display flight seemed to be mainly an advertising signal to potential mates. Display rate, as well as proportion of time spent in aerial display, increased with decreasing male size. During aerial display, small males also performed costly hovering flights more often and for relatively longer than large males. These results support the aerial display hypothesis.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9398383 TI - Size and pairing success in Gammarus duebeni: can females be too big? AB - The crustacean Gammarus duebeni exhibits precopula mate guarding and size assortative pairing, in which larger males tend to pair with larger females. Size assortative pairing may result from sexual selection or natural selection (mechanical or loading constraints limiting the size of female that can be carried by the male). If loading constraints are important, large females should have lower pairing success than females of intermediate size as they will be less likely to encounter sufficiently large males capable of carrying them in precopula. We tested this hypothesis in a laboratory study. Female pairing success was dependent on size; however, the relationship was curvilinear: pairing success increased with size up to a point, but larger females suffered decreased pairing success. This supports the hypothesis that loading constraints play a part in structuring size-assortative pairing in this species. We found no evidence for size-related female resistance in structuring the pattern of pairing. We considered size-related pairing success with regard to environmental sex determination and parasitic sex-ratio distortion in G. duebeni1997 The Association for the Study of Animal BehaviourCopyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9398384 TI - Cooperative signalling between opponents in fish fights AB - Cichlids of the species Nannacara anomala employ several colour displays during fights which do not seem to signal either fighting ability or motivation. How should these colour displays be interpreted when winning is reliably predicted by weight asymmetries? Medial Line colour displays were associated with, and predicted, tail-beating, while Vertical Bar colour displays were associated with mouth-wrestling. I suggest that these colour displays are used to facilitate the transmission of assessment information within a fight, and that they are an example of cooperative signalling between opponents. The results support the idea that the structure of fights contains strong cooperative aspects.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9398385 TI - Foraging costs in social carnivores AB - No abstractCopyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9398386 TI - Cooperative hunting and group size: assumptions and currencies AB - No abstractCopyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9398387 TI - A perspective on the neurobiology of language. AB - This paper provides a perspective on current issues and challenges in the investigation of the neurobiology of language. It is proposed that the speech/language deficits of aphasic patients reflect impairments in the processing components involved in accessing language. More specifically, it is hypothesized that many of these deficits result from changes in the activation level of word candidates in the lexicon. Because word recognition and lexical access processes are crucially involved in virtually all aspects of language processing, such an impairment has repercussions throughout the components of the linguistic grammar. It is suggested that the intersection of such language behaviors with the identification of underlying neural systems will define future research directions. Methodological and technological issues are discussed as they impact on current and future research. PMID- 9398388 TI - Phonological, semantic, and mediated priming in aphasia. AB - An auditory lexical decision task was conducted to examine rhyme, semantic, and mediated priming in nonfluent and fluent aphasic patients and normal controls. Overall, monosyllabic word targets were responded to faster when preceded by rhyming word and nonword primes than unrelated primes. Similarly, semantically related primes facilitated lexical decisions to word targets. No evidence of mediated priming emerged. Results for individual subjects suggest differences in patterns across the subject groups. Implications of the findings for the integrity of lexical access in aphasic patients are considered. PMID- 9398389 TI - Levels of morphological deficit: indications from inflectional regularity. AB - A language impairment that affects the production of inflected and/or derived words may result from a deficit that specifically affects morphological processing mechanisms, but it might also arise from whole-word processing failures as well (Badecker & Caramazza, 1987; Funnell, 1987). However, to motivate a true morphological impairment, the deficit must be understood in terms of one or more different levels of morphological structure. Minimally, we can distinguish a word's morphosyntactic representation from its morphophonological representation. In the single-case study reported here a deficit affecting the representation or processing of morphosyntactic representations is motivated. A critical part of the argument is that the deficit affects both regular and irregular inflection, and that no whole-word processing deficit can account for the particular pattern observed in this patient. PMID- 9398390 TI - Primary progressive aphasia: a review of 112 cases. AB - Primary progressive aphasia (PPA) was first recognized by Mesulam in 1982. Although dozens of cases have since been described, it has been difficult to place these cases into a coherent framework due to the wide variation in measures which have been reported. We review 170 contacts with 112 patients to provide a clinical, neuroanatomical, and neuropsychological profile of patients with the disorder. The progression of the disease is analyzed over a 10-year reporting period starting from symptom onset to show how progression affects five general linguistic skills: oral and written naming, reading, repetition, and general comprehension. The pattern of functional and neurological deficits in PPA is heterogeneous. Differences in the distribution of neurological anomalies between patients with bilateral and unilateral changes suggest that there may be two separate disease processes involved. PMID- 9398391 TI - Category-specific naming deficit for medical terms after dominant thalamic/capsular hemorrhage. AB - Postmortem, retrograde degeneration, and electrical stimulation studies have implicated the anterior pulvinar in language processing. We examined a patient who, after a hemorrhage affecting the dominant pulvinar and internal capsule, exhibited a circumscribed anomia for medical items and conditions. No other language disturbance was noted. Five category-specific word lists, matched for word frequency, were administered in a naming-to-definition format. Results indicated that the patient exhibited a significant category-specific naming deficit for medical items and conditions compared to matched control subjects. Although medical item lists were found to differ from nonmedical item lists in imageability and abstractness, B.C.'s category-specific deficit did not seem to be caused by word frequency, concept familiarity, imageability, or abstractness. Nor could the patient's performance be explained on the basis of deficits in broader semantic classifications (i.e., animate vs inanimate or man-made vs natural). The patient was unable to retrieve medical items even when given phonemic cues for those he could not name. Findings indicate that subtotal damage in the dominant pulvinar may create category-specific deficits. PMID- 9398392 TI - Lexical acquisition in probable Alzheimer's disease. AB - Patients with probable Alzheimer's disease (pAD) were exposed to a new verb in a naturalistic fashion. We probed their knowledge of the word's semantic and grammatical characteristics for several minutes following this exposure, and compared this with their performance on parallel measures assessing known words. Significant differences were seen between pAD patients and controls in the acquisition of the new verb's semantic meaning and its argument structure, but pAD patients did not differ from controls in the acquisition of the new word's grammatical form class. Individual patient analyses demonstrated parallel deficits restricted to the semantic meaning and argument structure of the new word and known words in several pAD patients, suggesting that a selective language impairment contributed to their word learning deficit. This pattern is consistent with an intimate relationship between semantic meaning and argument structure in semantic memory. Other pAD patients had difficulty learning about all aspects of the new word, despite good performance with known words, suggesting that compromised memory may have limited their lexical acquisition. PMID- 9398393 TI - Cerebral asymmetries in processing strategies for letter and symbol trigrams. AB - Several studies have shown that laterally presented consonant-vowel-consonant (CVC) strings produce both superior performance, and a more wholistic processing strategy in the right visual field/left hemisphere (RVF/LHEM), and a more sequential strategy in the inferior left visual field (LVF). To determine whether these strategies are applied to other types of trigrams subjects (n = 30) were asked to identify consonant and symbol trigrams briefly projected unilaterally to the LVF or RVF, or bilaterally (the same trigram in both fields--BVF). A second group of subjects (n = 30) first practiced pronouncing consonant trigrams and then viewed them tachistoscopically. Both tasks yield RVF advantages. Symbols are processed more wholistically in the LVF, more sequentially in the RVF and in an intermediate pattern when presented bilaterally. In contrast, subjects seem to chunk letters as bigrams, and do so equally well in all fields, and visual field differences in strategies emerge for consonants only when they are pronounced. Pronounceability of consonant trigrams, assessed with ratings and vocal reaction times, was predicted by orthographic regularity. Since the RHEM has limited phonetic skills, but it, like the LHEM, is privy to information on orthographic regularity, the error pattern on consonant strings indicates non-phonetic processing, whereas the RVF wholistic strategy for consonant-vowel-consonant strings appears to reflect phonetic processing. PMID- 9398394 TI - Image and language in human reasoning: a syllogistic illustration. AB - Existing accounts of syllogistic reasoning oppose rule-based and model-based methods. Stenning and Oberlander (1995) show that the latter are isomorphic to well-known graphical methods, when these are correctly interpreted. We here extend these results by showing that equivalent sentential implementations exist, thus revealing that all these theories are members of a family of abstract individual identification algorithms variously implemented in diagrams or sentences. This abstract logical analysis suggests a novel individual identification task for observing syllogistic reasoning processes. Comparison of the results of this task with the Standard Task confirms that the tasks are psychologically closely related, throwing light on sources of error, on subjects' sensitivity to metalogical properties, and on term-ordering phenomena. Since it avoids posing the subtask of formulating a quantified conclusion, the new task allows comparison of explanations of problem difficulty in terms of the number of models (e.g., Johnson-Laird & Bara, 1984) with alternatives in terms of the difficulty of choosing a quantifier for the conclusion. Logical concepts of source and conditional premisses provide a comprehensive account of term order data, including figural effects, at a level abstract with regard to imagistic or sentential representations. These results argue that much richer empirical evidence will be required to discriminate phenomenologically distinct reasoning processes than has hitherto been supposed. PMID- 9398396 TI - What Does Nonword Repetition Measure? A Reply to Gathercole and Baddeley PMID- 9398395 TI - Symmetry and asymmetry of human spatial memory. AB - Six experiments investigated the limiting conditions on and the causes of asymmetries in estimates of euclidean distance. Participants estimated distances between locations on recently learned maps or between buildings on their college campus. Estimates between landmarks and neighboring nonlandmarks were often asymmetric, but estimates between other pairs of locations were typically symmetric. These and other results were inconsistent with the predictions of models that attribute asymmetries to stimulus or to retrieval bias. A contextual scaling model of asymmetry is proposed. According to this model, asymmetries in proximity judgments are caused by general principles of human memory and judgment: (a) Stimuli differ in the contexts they establish in working memory and (b) magnitude estimates are scaled by the context in which they are made. PMID- 9398397 TI - A new concept of immune specificity emerges from a consideration of the self nonself discrimination. AB - The necessity to make a Self(S)-NonSelf(NS) discrimination is the evolutionary selection pressure for specificity of the immune response. A new definition of paratopic specificity, which is heuristic and generalizable, can be derived from an understanding of this selection pressure. Specificity of the paratope is defined by a Specificity Constant, K, which is the probability that a functional change in recognition will be anti-Self. In an antigen-unselected population, K is the proportion of cells that are anti-Self. This definition is unique in that it is derived from the function upon which evolution selects, namely the effector output. This paper describes how the concept of a Specificity Constant was derived, how it is estimated, what it can be used to explain, and how it impacts on repertoire and effectiveness of response. PMID- 9398398 TI - Circulating levels of IL-10 are critically related to growth and rejection patterns of murine mastocytoma cells. AB - Previously tumorigenic P815 tumor cells are rejected by histocompatible mice after transfection with a mutated retroviral gene, and the host is made resistant to subsequent challenge with tumorigenic (control) cells transfected with the nonmutated sequence. To functionally characterize the class I-restricted response to the tumor cell vaccine, we have assessed the in vitro (by CD8+ cells) and in vivo production of type 1 or type 2 cytokines in mice injected with either type of transfected P815 derivative. IL-12 and IL-10 were selectively or preferentially expressed by the regressor mice, and this correlated with the detection of functional type 1 reactivity in vivo (i.e., delayed-type hypersensitivity). Other cytokines were produced by the regressor mice only in vitro (IFN-gamma) or were not detected at all with either type of tumor recipient (IL-4). By means of monoclonal antibody-mediated neutralization or enhancement of endogenous cytokine levels, IL-10 was found to serve an important role in the growth and rejection patterns of the transfected P815 derivatives. In addition to previous evidence for an IL-12 requirement in promoting anti-P815 reactivity, these data establish IL-10 as an important cytokine in permitting optimal expression of this reactivity, which apparently develops in the absence of a strong bias toward a type 1 or type 2 cytokine response. PMID- 9398399 TI - Cross-linking of protein S bound to lymphocytes promotes aggregation and inhibits proliferation. AB - Recently, we reported that expression of the anticoagulant protein S is IL-4 inducible in primary T cells, and that protein S inhibits lymphoid cell procoagulant activity. Here, using a flow cytometric assay, we demonstrate that protein S binds to the surface of B and T lymphocytes. In addition, we show that cross-linking of protein S bound to lymphocytes induces aggregation and inhibits growth in cultures of primary B and T lymphocytes. Thus, our studies suggest that protein S is an IL-4-inducible T cell product that can affect B and T cell growth and aggregation via a lymphocyte protein S receptor. Interestingly, protein S joins thrombin and factor Xa as coagulation factors that modulate lymphocyte activation, suggesting that the clotting pathway may regulate wound-related inflammatory responses. PMID- 9398400 TI - Prolonged allogeneic and xenogeneic microchimerism in unmatched primates without immunosuppression by intrathymic implantation of CD34+ donor marrow cells. AB - Engraftment of stem cell-enriched donor marrow implanted in the thymus of a foreign host might facilitate acceptance of donor-specific organ or tissue grafts. To test this hypothesis, allogeneic and xenogeneic CD34+ marrow cells from unrelated adult male baboons and humans were injected intrathymically in eight infant female baboons, both with and without standard cyclosporine-based immunosuppression. In allogeneic experiments, male (donor) cells, of both T- and B-cell lineages, were detected by PCR in the peripheral blood of all six recipients and persisted for at least 15 months in 2/4 recipients studied longtutudinally. Donor-derived skin grafts survived twice as long as third party grafts in unimmunosuppressed recipients. In xenogeneic protocols, human male (donor) cells were demonstrable for 7 and 15 months, respectively, in two baboon recipients with evidence that implanted human CD34+ cells had produced lymphoid progeny. Survival of donor-specific skin xenografts was prolonged in one of two recipients. These experiments demonstrate that the intrathymic injection of CD34+ marrow cells can result in long-lasting lymphohematopoietic microchimerism in unrelated primates even without immunosuppression and can alter donor-specific skin graft survival. PMID- 9398401 TI - Ligation of CD40 potentiates Fas-mediated activation of the cysteine protease CPP32, cleavage of its death substrate PARP, and apoptosis in Ramos-Burkitt lymphoma B cells. AB - The proliferation and survival of a B cell population is necessarily tightly controlled to prevent the arisal of malignancy or autoimmunity. The expansion or elimination of a B cell clone is determined through a complex interaction of the tumour necrosis factor receptor/nerve growth factor receptor family members CD40 and Fas, which are expressed on the B cell surface, with their respective physiological ligands (CD40L and FasL) expressed on the surface of CD4+ T cells. The regulation of B cell growth by signals transduced through CD40 and Fas contributes to the maintenance of peripheral tolerance and likely takes place and in the germinal centres (GC) of secondary lymphoid tissues. In this study, we investigate the relationship between the expression of Fas and B cell survival following engagement of CD40 and Fas in the Epstein-Barr virus-genome-negative Ramos-Burkitt lymphoma (Ramos-BL) B cell line model of GC B lymphocyte selection during maturation of the humoral immune response. We now present evidence that Ramos-BL B cells constitutively express both Fas and FasL on their surface and that expression of Fas, but not FasL, is enhanced following ligation of CD40. Coligation of CD40 and Fas, triggers for growth inhibition, activation of the interleukin-1 beta-converting enzyme, now caspase, family member CPP32 (caspase 3) but not Ich-1L (caspase-2), cleavage of its death substrate poly(ADP-ribose) polymerase, and apoptosis from the G1 phase of cell cycle; engagement of Fas alone fails to trigger for growth inhibition and apoptosis but enhances AgR mediated cellular death. This CD40-potentiated Fas-triggered growth inhibition and apoptosis occurs in the presence of CD40-induced expression of the anti apoptotic proteins Bcl-xL and Bcl-2. Taken together, these data indicate that ligation of CD40 facilitates efficient coupling of Fas to the caspase-mediated pathway of apoptosis. PMID- 9398402 TI - Influence of retinoic acid on the differentiation pathway of T cells in the thymus. AB - This study investigated the ability of retinoic acid (RA) to influence T cell differentiation. All-trans-RA had marked effects on T cell differentiation in murine fetal thymic organ cultures (FTOCs). The time course of the effect of all trans-RA in FTOC of day 14 C57BL/6 embryos revealed a twofold increase in the frequency of CD4 single-positive (SP) cells and a high level of CD3-bearing cells (CD3high cells) at a later stage of T cell development. At an earlier stage, all trans-RA induced a twofold increase in the frequency of CD4 SP cells, but significantly suppressed the upregulation of CD3 and TCR. Reverse transcription PCR using RA receptor (RAR) subtype-specific primers showed that RAR alpha but not beta and gamma is expressed during T cell development in the thymus and that its expression was associated with the generation of CD4/CD8 double-positive (DP) cells. In FTOC of day 16 BALB/c embryos, the level of V beta 3high cells was greatly reduced (1.4% of the CD3high cells) in response to the mouse mammary tumor virus-6-encoded superantigen, but V beta 3-bearing cells were rescued from the deletion in the presence of all-trans-RA (5.6% of the CD3high cells). Further, the inhibitory effect of all-trans-RA on thymocyte deletion was observed when the deletion was induced by a low concentration of staphylococcal enterotoxin B in FTOC. Taken together, these data suggest that RA increases the frequency of mature and self-reactive T cells in the thymus, possibly by inhibiting the process of negative selection at the DP stage of T cell differentiation. PMID- 9398403 TI - Selective antigen presenting activity of cortical thymic epithelial cells against CD4+ T cells associated with both lack of costimulatory molecules and inefficient presentation of MHC-peptide ligands. AB - Selective activation among several effector functions of a T cell clone, DB14, specific for pigeon cytochrome c 43-58 (p43-58) and restricted to I-Ab/d was induced by antigen (Ag) presentation with nonprofessional Ag-presenting cells (APC), cortical thymic epithelial cells (c-TEC) (B7-1- CD40-), whereas full activation of the DB14 was induced with another nonprofessional APC, I-Ab L cell (B7-1+ CD40+). In the present study, to elucidate the mechanism underlying the selective activation of DB14 cells by c-TEC, we established c-TEC transfected with human CD40 alone (huCD40-c-TEC) or both human CD40 and murine B7-1 (huCD40/mB7-1-c-TEC), and compared the APC functions with those of the original c TEC and I-Ab L cell. IFN-gamma production but not the proliferative response of DB14 was elevated by Ag presentation with huCD40-c-TEC as compared with unmanipulated c-TEC. On the other hand, upon stimulation with Ag plus huCD40/mB7 1-c-TEC both a significant proliferative response and IFN-gamma production were induced in DB14. However, the level of these responses did not reach that induced in the presence of I-Ab L cell. A similar pattern of APC functions was demonstrated with the other B7-independent T cell clone, PAB3, or T cell hybridomas (DBhy22 and BD7-5) which are basically independent of costimulation for the activation. The present finding along with our previous report that several structural differences of I-Ab molecules are present between c-TEC and I Ab L cell suggests that the distinct APC activity of c-TEC is attributable not only to a lack of B7-1 and CD40 but also to inefficient presentation of MHC peptide complex on the c-TEC. PMID- 9398404 TI - IL-2 and IL-7 induce heterodimerization of STAT5 isoforms in human peripheral blood T lymphoblasts. AB - Despite differences in T cell responses induced by interleukin (IL)-2 and IL-7, both cytokines modulate T cell functions by activation of signal transducers and activators of transcription (STAT) proteins. We examined the contribution of the two isoforms of STAT5, STAT5A and STAT5B, to IL-2- and IL-7-induced activation of human peripheral blood T lymphoblasts. Both cytokines induced assembly of STAT5A and STAT5B containing complexes capable of binding to the interferon-gamma activation sequence (GAS), and these complexes rapidly translocated (within 1 min) into the nucleus of IL-2- or IL-7-treated cells. The kinetics of this translocation were delayed in IL-7-treated as compared to IL-2-treated cells. IL 2 and IL-7 were equivalent in their ability to induce tyrosine phosphorylation of STAT5A and STAT5B and to facilitate binding of these STATs to an immobilized GAS element. Both IL-2 and IL-7 induced substantial amounts of STAT5A/STAT5B heterodimerization. Moreover, we observed constitutive association of STAT3 with each STAT5 isomer. These data suggest that IL-2 and IL-7 induce assembly of STAT heterodimers in a similar manner and that subsequent cellular responses may be driven by induction of similar sets of genes. PMID- 9398405 TI - Suppression of human B cell responsiveness by CD4+ T cells does not involve CD95 CD95 ligand interactions. AB - Although human CD4+ T cells have been shown to regulate humoral immune responses by directly inhibiting B cells, the precise sequelae for the mechanism of suppression have not yet been delineated. The present study was therefore designed to explore the nature of T cell-B cell collaboration to suppress B cell responses. Special attention was directed to the roles of Fas (CD95)-Fas ligand (FasL) interactions. The suppressive activity was assessed by the effects of mitomycin C-untreated CD4+ T cells activated by immobilized anti-CD3 for 72 h (CD4+ suppressors) on the production of IgM and IgG of B cells stimulated for 72 h with immobilized anti-CD3-activated mitomycin C-treated CD4+ T cells. In this model system, B cells stimulated with anti-CD3-activated mitomycin C-treated CD4+ T cells expressed functional Fas receptors. Accordingly, addition of anti-Fas mAb CH-11 inhibited the cluster formation and differentiation of activated B cells as a result of apoptotic cell death in a manner that was completely reversed by a neutralizing anti-Fas mAb ZB4. However, neither ZB4 nor anti-FasL mAb reversed the suppression of B cell responses by anti-CD3-induced CD4+ suppressors. Of interest, ZB4 significantly enhanced the production of IgM and IgG induced by anti-CD3-activated mitomycin C-treated CD4+ T cells in the absence of CD4+ suppressors. Consistently, mitomycin C-treated CD4+ T cells as well as mitomycin C-untreated CD4+ T cells expressed comparable levels of FasL upon activation with immobilized anti-CD3, although their intensities were very modest. These results indicate that B cells activated with anti-CD3-stimulated CD4+ T cells express functional Fas receptors and are sensitive to Fas-mediated apoptosis. However, the data also suggest that interactions other than Fas-FasL may play a critical role in direct cellular collaboration between activated B cells and anti-CD3 induced CD4+ suppressors to inhibit B cell responses. PMID- 9398406 TI - Surface Coverage and Its Determination: Role of Acid-Base Interactions in the Surface Treatment of Mineral Fillers AB - The role of acid-base interactions in the nonreactive surface treatment of mineral fillers was studied by a dissolution method. The effect of treatment on the surface properties of CaCO3 covered with stearic acid was determined by ESCA measurements and by the calculation of the basicity of the treated fillers. The dissolution measurements were carried out in twelve different solvents. The maximum amount of surfactant adsorbed on the filler surface (cmax) proved to be independent of the acid-base character of the solvent, while the amount bonded proportionally (c100) depended strongly on it. The results showed that competitive processes determine the surface coverage of the filler: the adsorption of the surfactant and the solvent on the filler surface competes with the solution of the treating material in the solvent. Sedimentation experiments gave valuable information about the strength of solvent/filler interaction. The surface characteristics of the filler changed with surface coverage; the surfactant concentration at which basicity attains its minimum agrees well with the monolayer coverage determined by ESCA and with the c100 value obtained in apolar solvents. The results have practical relevance as well, similar competitive solution/adsorption processes may take place also in composites containing surface treated fillers. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9398407 TI - Studies on Interaction of Dodecyltrimethylammonium Bromide with Na- and Al Montmorillonite AB - The adsorption isotherms and adsorption enthalpies of dodecyltrimethylammonium bromide (DDTMAB) on Na- and Al-montmorillonite were determined in the pH range 2 12. The basal spacings of the clays were also studied by X-ray diffraction. Interactions of DDTMAB with the two montmorillonites predominate mostly through cation exchange in the pH range studied. In the case of Na-montmorillonite, the amount of adsorption increases with pH, but adsorption enthalpy decreases with pH. For Al-montmorillonite, adsorption enthalpy exhibits nonmonotonic variation with pH, although the amount of adsorption increases with pH monotonously. The adsorption enthalpies of DDTMAB on the clays are negative. The adsorbed DDTMAB molecules in clay interlayers present a bilayer arrangement at saturation. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9398408 TI - Determination of the Surface Tension of Microporous Membranes Using Contact Angle Measurements AB - In this paper, a new method of determining the surface tension of the solid material that a microporous membrane is made from is introduced. The method is based on the well known determination of the so-called contact angle that is formed on the solid/liquid/gaseous three phase line. A nonideal state of the solid phase leads to a deviation of the contact angle that can be observed experimentally from the equilibrium angle that arises from the thermodynamically state of lowest energy, as it must be used to calculate the solid surface tension via the Young equation. The deviation caused from the porous structure of the solid material will be taken into account in this work. Doing so, we derived an equation that connects the surface porosity, the measured contact angle, and the equilibrium contact angle. Using this equation, the measured and therefore deviated contact angles can be corrected for the porosity of the solid material, yielding the contact angle observable on a surface made from the same but nonporous material. The equation derived was tested on different microporous membranes made from expanded poly(tetrafluoroethylene). The surface porosity needed was determined using scanning electron microscopy followed by computerized image analysis. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9398409 TI - Static Drops on an Inclined Plane: Equilibrium Modeling and Numerical Analysis AB - The continuum description of the equilibrium of small liquid drops located on a sloping plane is still discussed. The effect of drop holdup on the contact surface is modeled by describing the counteraction of a possible rolling liquid flow. This paper studies numerically the effect of the contact angle hysteresis, the critical slope angle at which the drop flows out. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9398410 TI - Nonideal Mixing in Adsorbed Film and Micelle of Ionic-Nonionic Surfactant Mixtures AB - The surface tension of aqueous solutions of dodecylammonium chloride-octyl methyl sulfoxide (OMS) and dodecyltrimethylammonium bromide-OMS mixtures was measured as a function of the total molality of surfactants and the composition of OMS. The results were analyzed according to our thermodynamic procedure; the phase diagrams of adsorption and micelle formation and the diagram illustrating the composition relation between the micelle and the adsorbed film at the critical micelle concentration were drawn. The activity coefficient and excess Gibbs energy in the adsorbed film and the micelle were defined and then evaluated from the phase diagrams. They were found to give useful information on the deviation from ideal mixing and intermolecular interaction in the adsorbed film and micelle. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9398411 TI - Small-Angle X-Ray Scattering Study of the Formation of Colloidal Silica Particles from Alkoxides: Primary Particles or Not? AB - The formation of colloidal silica particles and the dynamics of the nanostructure of the particles are investigated by small-angle X-ray scattering (SAXS) technique. Solute concentrations of 0.5 M tetraethylorthosilicate (TEOS), 1.1 or 2.2 M water (H2O), and 0.04 or 0.1 M ammonia base (NH3) in ethanol were used to obtain reaction conditions as close to those of the Stober method as possible and to have reaction kinetics that were slow enough to probe the changes in the nanostructure of the growing particles and to obtain good statistics from the SAXS measurements. We measured the changes in the radius of gyration and the fractal dimension as a function of time during growth. Remarkably, we find that, after an induction period, the first particles to appear in the solution have a radius of gyration of approximately 10 nm and are mass fractals characterized by their polymeric, open structure. This stage is followed by an intraparticle densification process and smoothing of the interface, leading to the usual compact nonfractal, stable structures. The growth models proposed so far cannot account for the observed continuous changes of stages during the formation and growth of the particles. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9398412 TI - Spectroscopic Studies of Dextrin Adsorption onto Colloidal ZnS AB - The interaction of dextrin with colloidal ZnS has been investigated through adsorption studies and FT-IR spectroscopy in the 4000-400 cm-1 range. The adsorption capacity is estimated to be around 1 mg/m2. Maximum adsorption is found to be constant below pH approximately 7 and to increase with pH at least up to pH 11. Eighty percent of maximum adsorption is achieved within 3 min after addition of the dextrin. Based upon FT-IR studies and titration data, an adsorption mechanism is proposed. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9398413 TI - Structural and Dynamic Properties of Lecithin-Alcohol Based w/o Microemulsions: A Luminescence Quenching Study AB - Water-in-oil microemulsions have been made with lecithin in the presence of some alcohols. The structure of the microemulsions has been studied by steady-state and time-resolved analysis of the luminescence quenching of Ru(bipy)32+ by Fe(CN)63-. We found that well-defined microemulsions can be made only in the presence of short-chain alcohols such as propanol-1 and butanol-1. There exists a threshold for water content in order to obtain typical reverse micelles. Thus in the case of propanol-1, water/surfactant ratio wo should be above 20. By varying water content in the range 20 < wo IP(O) > IP(N), though in the case of CO adsorption, the amounts adsorbed on IP(A) and IP(O) were not greatly different. In all cases the amounts adsorbed represented only fractional coverage. Adsorption of the more acidic CO2 is thought to be favored more by basic Ox-2 than by O2- sites on both IP(O) and IP(A), but with surface hydroxyl groups also playing a role (particularly on IP(A)). The CO2 adsorption should result in the formation of mono-, di-, and polydentate carbonate and bicarbonate species, with increasing degassing temperatures favoring the polydentate species and the decomposition of the bicarbonate and carbonate to form undissociated CO2. The adsorption of CO (a weak base) is postulated to take place on strong Lewis acid, highly coordinated, metal sites to form metal carbonyl species, on strong base sites (O2-) to form carbonite, oxalate, and ketenic species, and, to a lesser degree, on surface hydroxyl groups to form formyl and formate species. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9398432 TI - Mixed cultures of avian blastoderm cells and the quail mesoderm cell line QCE-6 provide evidence for the pluripotentiality of early mesoderm. AB - During the early stages of embryogenesis, the mesoderm gives rise to cells of the cardiovascular system which include cardiac myocytes and vascular endothelial and red blood cells. We have investigated the development of these cell phenotypes using aggregate cultures of avian blastoderm cells, which replicated mesodermal cell diversification. The cell phenotypes expressed by the blastoderm cells were dependent upon the age of the blastoderm cells, with Hamburger-Hamilton stage 3 or 4 cells giving rise to endothelial and red blood cells and stage 5 cells producing endothelial and myocardial cells. To begin to understand the stage dependency of the cellular diversification of these aggregate cultures, we treated the cultures with various signaling factors that have been shown to be present in the early avian embryo. These experiments showed that stem cell factor and TGF alpha altered cell phenotypes by stimulating red blood cell and myocardial differentiation, respectively. The ability of these growth factors to shift the differentiation profile of aggregate cultures demonstrated the plasticity of early embryonic cells. To explore the diversification of individual mesodermal cells, labeled QCE-6 cells were incorporated within these blastoderm aggregate cultures. Previous studies have shown that this quail mesodermal cell line possesses characteristics of early nondifferentiated mesodermal cells and can be induced to express either myocardial or endothelial cell phenotypes (C. A. Eisenberg and D. M. Bader, 1996, Circ. Res. 78, 205-216). In the present study, we show that when these cells were cultured as a component of blastoderm cell aggregates, they differentiated into fully contractile cardiomyocytes or endothelial or red blood cells. Moreover, QCE-6 cell differentiation was in accordance with that displayed by the blastoderm cells. Specifically, QCE-6 cells differentiated into red blood cells when cultured within stage 3 or stage 4, but not stage 5, blastoderm cell aggregates. Accordingly, the differentiation of QCE 6 cells into beating cardiomyocytes only occurred when these cells were incorporated into stage 5 blastoderm cell aggregates. The identical sorting and differentiation patterns that were exhibited by QCE-6 and blastoderm cells suggest that expression of differentiated cell types within the early mesoderm is directed by the surrounding environment without immediate cellular commitment. In addition, these results provide further evidence that QCE-6 cells are representative of a multipotential mesodermal stem cell and that they possess the potential to exhibit fully differentiated cell phenotypes. PMID- 9398433 TI - MAP and cdc2 kinase activities at germinal vesicle breakdown in Chaetopterus. AB - In frog oocytes, activation of mitogen-activated protein kinase (MAPK, ERK) leads to activation of cdc2 and germinal vesicle breakdown (GVBD). By contrast, in starfish, MAPK is activated after GVBD. Here we have examined the relative involvements of MAPK and cdc2 in GVBD of Chaetopterus oocytes. MAPK was rapidly tyrosine-phosphorylated and activated (within 1-2 min) in response to exposure of the oocytes either to natural seawater (the normal trigger of GVBD in this organism) or to the tumor-promoting phorbol ester 12-O-tetradecanoylphorbol 13 acetate (TPA), which can also elicit GVBD. This response preceded the tyrosine dephosphorylation and activation of cdc2 by several minutes. MAPK phosphorylation and activation were transient, lasting only until GVBD occurred and the spindle migrated to the cortex. The enzyme was not phosphorylated again as a result of egg activation. These results are consistent with the hypothesis that the activation of MAPK has a role in GVBD. However, PD 98059, a potent and selective inhibitor of MEK, the protein kinase that phosphorylates and activates MAPK, blocked the phosphorylation of MAPK but did not block GVBD, the dephosphorylation and activation of cdc2, or spindle formation and migration. Oocytes that underwent GVBD in PD 98059 could be fertilized and cleaved normally. Ionophore A23187, although it caused germinal vesicles to disappear and caused transient phosphorylation of MAPK, did not cause dephosphorylation of cdc2, and therefore this disappearance is artifactual. These results suggest that MAPK activation is neither obligatory nor sufficient for either GVBD or meiotic metaphase arrest in Chaetopterus and that activation of MAPK and cdc2 occur on independent, parallel pathways. PMID- 9398434 TI - A critical period of ear development controlled by distinct populations of ciliated cells in the zebrafish. AB - The zebrafish (Danio rerio) is a useful model system for analyzing development of the inner ear. A number of mutations affecting the inner ear have been identified. Here we investigate the initial stages of otolith morphogenesis in wild-type embryos as well as in monolith (mnl) mutant embryos, which fail to form anterior otoliths but otherwise appear normal. Otolith growth is initiated at 18 18.5 h by localized accretion of free-moving precursor particles. This process, referred to as otolith seeding, is regulated by two classes of cilia: First, kinocilia of precociously forming hair cells (tether cells) bind seeding particles, thereby localizing otolith formation. Tether cells usually occur in pairs at the anterior and posterior ends of the ear. Despite the presence of functional kinocilia, tether cells initially appear immature and do not acquire the characteristics of mature hair cells until approximately 21.5 h. Second, beating cilia distributed throughout the ear agitate seeding particles, thereby inhibiting premature agglutination. Constraining particles with laser tweezers caused them to fuse into large untethered masses. Bringing such masses into contact with tethered otoliths caused them to fuse, greatly enhancing otolith growth. Selectively enhancing one otolith greatly inhibited growth of the second, creating an imbalance that persisted for many days. Seeding particles and beating cilia disappear soon after 24 h, and the rate of otolith growth decreases by nearly 90%. In mnl mutant embryos, tethers and beating cilia are distributed normally, but anterior otoliths fail to form in 80-85% of mutant ears. The binding properties of seeding particles appear normal, as shown by their ability to fuse when entrapped by laser tweezers and their binding to posterior tethers. We infer that anterior tethers have a weakened ability to bind seeding particles in mnl embryos. Immobilizing mnl embryos with the anterior end of the ear oriented downward effectively concentrated the dense seeding particles near the anterior tethers and permitted all to form anterior otoliths. However, immobilizing mnl embryos after 24 h when seeding particles were depleted did not facilitate anterior otolith formation. Together, these data demonstrate that the ability to initiate otolith formation is limited to a critical period, from 18.5 to 24 h, and that interfering with the functions of tether cell kinocilia or beating cilia impairs otolith seeding and subsequent otolith morphogenesis. PMID- 9398435 TI - Mesoderm is required for the formation of a segmented endodermal cell layer in the leech Helobdella. AB - The homeobox gene Lox3 is expressed in a segmentally iterated pattern within the endoderm of the leech Helobdella. We use that expression here to study endoderm differentiation following experimental ablations of mesoderm. Lox3 RNA was first detected by in situ hybridization at the stage when a definitive cellular endoderm is formed from its syncytial precursor and was never observed in derivatives of other germ layers. Expression is initially distributed throughout the endoderm, but rapidly disappears from specific regions of the nascent gut wall so as to produce a pattern of segmental stripes. The stripe pattern differs markedly between midgut organs, with thin stripes of Lox3 expression in the intercaecal constrictions of the crop and wide stripes of Lox3 expression marking the caecal bulges of the intestine. Lox3 expression in the rectum is not obviously segmental. Ablation of segmental mesoderm in the early Helobdella embryo prevents the formation of definitive endoderm and the expression of Lox3 RNA and leads to abnormalities in the morphogenesis of the gut tube. These endodermal deficits are precisely coextensive with the zone of mesodermal deficiency, suggesting that the mesoderm normally acts to promote the formation of the endodermal cell layer via local cell interactions. The segmental pattern of Lox3 expression is largely unaffected in portions of the endoderm surrounding such deficits, suggesting that endodermal segmentation is not established by lateral interactions within that tissue layer. Rather, we propose that the segmental organization of the endoderm is imprinted by vertical interactions with the segmental mesoderm. PMID- 9398436 TI - The expression domain of two related homeobox genes defines a compartment in the chicken inner ear that may be involved in semicircular canal formation. AB - Homeobox-containing genes encode a class of proteins that control patterning in developing systems, in some cases by acting as selector genes that define compartment identity. In an effort to demonstrate a similar role for such genes during ear development in the chicken, we present a detailed expression study of two related homeobox-containing genes, SOHo-1 and GH6, using in situ hybridization. At otocyst stages the two genes define a broad lateral domain of expression, which may represent a developmental compartment. Three-dimensional computer reconstructions of SOHo-1 expression at these and later stages revealed that the lateral domain becomes progressively restricted to the three semicircular canals. Thus, SOHo-1 and GH6 are among a small group of markers for a specific structural component of the inner ear. The gene expression domain initially includes the sensory regions of the semicircular canals, known as the cristae ampullaris, but none of the other four sensory organs which were recognizable by BMP4 expression during early morphogenesis (stages 19-24). Significantly, two of the sensory organs (the superior and posterior cristae) were found at the limits, or boundaries, of the SOHo-1/GH6 expression domain, suggesting that compartment boundaries may be involved in specifying sensory organ location as well as identity. Maintained expression at the boundaries may aid in specifying the location of canal outgrowth. These concepts are presented as a formal model which emphasizes that patterning information could be provided at the boundaries of gene expression domains in the inner ear. PMID- 9398437 TI - Altered forebrain and hindbrain development in mice mutant for the Gsh-2 homeobox gene. AB - The patterning of the mammalian brain is orchestrated by a large battery of regulatory genes. Here we examine the developmental function of the Gsh-2 nonclustered homeobox gene. Whole-mount and serial section in situ hybridizations have been used to better define Gsh-2 expression domains within the developing forebrain, midbrain, and hindbrain. Gsh-2 transcripts are shown to be particularly abundant in the hindbrain and within the developing ganglionic eminences of the forebrain. In addition, mice carrying a targeted mutation of Gsh 2 have been generated and characterized. Homozygous mutants uniformly failed to survive more than 1 day following birth. At the physiologic level the mutants experienced apnea and reduced levels of hemoglobin oxygenation. Histologically, the mutant brains had striking alterations of discrete components. In the forebrain the lateral ganglionic eminence was reduced in size. In the hindbrain, the area postrema, an important cardiorespiratory chemosensory center, was absent. The contiguous nucleus tractus solitarius, involved in integrating sensory input to maintain homeostasis, was also severely malformed in mutants. Immunohistochemistry was used to examine the mutant brains for alterations in the distribution of markers specific for serotonergic and cholinergic neurons. In addition, in situ hybridizations were used to define expression patterns of the Dlx 2 and Nkx 2.1 homeobox genes in Gsh-2 mutant mice. The mutant lateral ganglionic eminences showed an abnormal absence of Dlx 2 expression. These results better define the genetic program of development of the mammalian brain, support neuromeric models of brain development, and further suggest similar patterning function for homeobox genes in phylogenetically diverse organisms. PMID- 9398438 TI - Cellular mechanism underlying neural convergent extension in Xenopus laevis embryos. AB - Convergent extension, the simultaneous narrowing and lengthening of a tissue, plays a major role in shaping and patterning the neural ectoderm in vertebrate embryos. In this paper, we characterize the cellular mechanism underlying convergent extension of the neural ectoderm in the Xenopus laevis late gastrula and neurula embryo. Neural ectoderm in X. laevis consists of two components, a superficial layer of epithelial cells overlying deep mesenchymal cells. To investigate the force contribution of the deep cells to convergent extension, we explanted single layers of neural deep cells from late gastrula stage embryos. These "neural deep cell explants" undergo active convergent extension autonomously, implying that these cells contribute force for neural convergent extension in vivo. Using time-lapse videorecording of these explants, we observed the neural deep cell behaviors (previously hidden behind an opaque epithelium) underlying convergent extension. We show that neural deep cells mediolaterally intercalate to form a longer, narrower tissue and that cell shape change and cell division contribute little to their convergent extension. Moreover, we characterize the neural deep cell motility driving mediolateral intercalation, also using time-lapse videorecordings. Analyses of these videos revealed that, on average, neural deep cells exhibit mediolaterally biased protrusive activity which is expressed in an episodic fashion. We propose that neural deep cells accomplish mediolateral intercalation by applying their protrusions upon one another, exerting traction, and pulling themselves between one another. This mechanism is similar to that previously described for convergent extension of the mesodermal cells. However, because the neural deep cells do not mediolaterally elongate during their convergent extension as the mesodermal cells do, we predict that a given intercalation will result in more extension for neural deep cells than for the mesodermal cells. Intercalation of neural cells also likely occurs in a more episodic manner than that of the mesodermal cells because the neural cells' mediolateral protrusive activity is episodic, whereas the protrusive activity of mesodermal cells is more continuous. These differences in protrusive activity and cell shape changes between the neural and mesodermal regions may reflect specializations of the same basic mechanism of mediolateral intercalation, tailored to accommodate other aspects of patterning and development of each tissue. These descriptions of the active cell motility underlying neural convergent extension in X. laevis are the first high-resolution video documentation of protrusive activity during neural convergent extension in any system. Our findings provide an important step in the investigation of neural convergent extension in X. laevis and further our understanding of convergent extension in general. PMID- 9398439 TI - Heterogeneous expression of multiple putative patterning genes by single cells from the chick hindbrain. AB - The metameric organization of the vertebrate hindbrain into rhombomeres appears to result from the patterned expression of several transcription factors and putative signaling molecules. We have applied a refined single-cell reverse transcription-polymerase chain reaction strategy to examine the molecular logic proposed to pattern the hindbrain at the single-cell level. This technique allows analysis of the concurrent expression of several genes within an individual cell at higher sensitivity than by in situ hybridization. Our results demonstrate that cells in rhombomere (r) 4 and r5 are heterogeneous in their expression of Hoxa-3, Hoxb-2, Sek-1, and Krox-20, suggesting that single cells are dynamically regulating their rhombomere-specific gene-expression profiles. Furthermore, the strong correlation between Sek-1 and Krox-20 expression at stage 12 was greatly diminished by stage 16, suggesting that the proposed interdependence of these two genes is present only at early stages of hindbrain development. PMID- 9398440 TI - Single-cell analysis of regulatory gene expression in quiescent and activated mouse skeletal muscle satellite cells. AB - Repair and regeneration of adult skeletal muscle are mediated by satellite cells. In healthy muscle these rare mononucleate muscle precursor cells are mitotically quiescent. Upon muscle injury or degeneration, members of this self-renewing pool are activated to proliferate and then differentiate. Here we analyzed in single satellite cells the expression of a set of regulatory genes that are candidates for causal roles in satellite cell activation, maturation, and differentiation. Individual cells were identified as satellite cells and selected for analysis based on their physical association with single explanted myofibers or their position beneath the basal lamina in unperturbed muscle tissue. Using a multiplex single-cell RT-PCR assay we simultaneously monitored expression of all four MyoD family regulators of muscle determination and differentiation (MRFs) together with two candidate markers of satellite cell identity, c-met and m-cadherin. By making these measurements on large numbers of individual cells during the time course of satellite cell activation, we were able to define which expression states (possible combinations of the six genes) were represented and to specify how the representation of each state changed with time. Activated satellite cells began to express either MyoD or myf5 first among the MRFs; most cells then expressed both myf-5 and MyoD simultaneously; myogenin came on later in cells expressing both MyoD and myf5; and many cells ultimately expressed all four MRFs simultaneously. The results for fiber-associated satellite cells from either predominantly fast or slow muscles were indistinguishable from each other. The c met receptor tyrosine kinase was also monitored because it is a candidate for mediating activation of quiescent satellite cells (Allen et al., 1995) and because it might also be a candidate molecular marker for satellite cells. A significant difficulty in studying mouse satellite cells has been the absence of molecular markers that could identify them in the quiescent state before expression of MRFs or desmin and distinguish them from fibroblasts. We show here that c-met receptor is present beneath the basal lamina on presumptive satellite cells in intact muscle and that c-met mRNA and protein are expressed by all myofiber-associated satellite cells from the time of explant through the course of activation, proliferation, and differentiation. c-met was not detected in muscle-derived fibroblasts or in other mononucleate cells from healthy muscle explants. When compared directly with m-cadherin, which has previously been suggested as a marker for quiescent satellite cells, m-cadherin mRNA was detected only in a small subset of satellite cells at early times after myofiber explant. However, at late times following activation (by 96 hr in this fiber culture system), c-met and m-cadherin were uniformly coexpressed. From the individual satellite cell expression types observed, a model of the satellite cell population at rest and during the time course of activation was generated. PMID- 9398441 TI - A Drosophila kinesin-like protein, Klp38B, functions during meiosis, mitosis, and segmentation. AB - We show that klp38B, isolated as a mutation that dominantly prolongs blastoderm mitotic cycles in Drosophila, encodes a Drosophila kinesin-like protein. Further genetic analyses show that Klp38B not only functions during mitosis, but is also required for meiosis and abdominal segmentation. Sequence comparisons suggest that Klp38B encodes an amino-terminal microtubule motor domain, a central alpha helical coiled-coil domain, and a C-terminal globular domain. Evidence that Klp38B is required during meiosis is that flies transheterozygous for mutations in both klp38B and nod have a high frequency of 4th chromosome meiotic nondisjunction. Nod is a chromokinesin, a chromosome binding kinesin, that is believed to provide astral-exclusion forces during the metaphase stage of meiosis. Evidence that Klp38B is required during mitosis is that embryos from female germline clones of klp38B mutations have holes in the cuticle similar to a zygotic string (dCDC25) phenotype. Also, anti-Klp38B antibody injection into precellularization blastoderm embryos causes developmental arrest and the formation of circular mitotic figures. We speculate, based on these phenotypes, that Klp38B is a chromokinesin that provides astral-exclusion forces on the chromosomes during meiosis and mitosis. Consistent with this hypothesis, we have identified an HMG-1 homologous region on Klp38B that could potentially bind AT rich DNA sequences. Finally, we show that klp38B mutations have defects in abdominal segmentation, suggesting that Klp38B, like Xenopus chromokinesin Xklp1, might be involved in polar granule formation. PMID- 9398442 TI - ARNT-deficient mice and placental differentiation. AB - We used homologous recombination in embryonic stem cells to generate mice heterozygous for an aryl hydrocarbon nuclear translocator (ARNT) null mutation. These mice were intercrossed, but no live homozygous Arnt-/- knockout mice were produced among 64 newborns. Homozygotes die in utero between 9.5 and 10.5 days of gestation. Abnormalities included neural tube closure defects, forebrain hypoplasia, delayed rotation of the embryo, placental hemorrhaging, and visceral arch abnormalities. However, the primary cause of lethality appears to be failure of the embryonic component of the placenta to vascularize and form the labyrinthine spongiotrophoblast. This may be related to ARNT's known role in hypoxic induction of angiogenesis. We found no defects in yolk sac circulation. PMID- 9398443 TI - NGF-mediated survival depends on p21ras in chick sympathetic neurons from the superior cervical but not from lumbosacral ganglia. AB - In rat embryonic sympathetic neurons from the superior cervical ganglia (SCG) NGF mediated survival depends on the activation of the trkA receptor tyrosine kinase and on the activity of the intracellular plasmamembrane-anchored small G-protein p21ras. In contrast, chick sympathetic neurons derived from the more caudally located lumbosacral chain ganglia (LSCG) do not respond to activated p21ras (G12V Ha-ras mutant). In these neurons endogenous p21ras and its downstream effector MAP kinase are activated but are not essential for NGF-dependent survival. Here we show that also in chick sympathetic neurons of the SCG permanently activated p21ras protein does promote neuron survival. Consistently, their NGF-mediated survival is sensitive to Fab fragments blocking endogenous p21ras activity. These results suggest that sympathetic neurons derived from sympathoenteric (SCG) and sympathoadrenal (LSCG) lineages differ in their requirement for p21ras in the NGF mediated survival pathways. PMID- 9398444 TI - Glial changes in the phrenic nucleus following superimposed cervical spinal cord hemisection and peripheral chronic phrenicotomy injuries in adult rats. AB - The objective of the present study was to characterize the microglial and astroglial reaction in the phrenic nucleus following either an ipsilateral C2 spinal cord hemisection, a peripheral phrenicotomy, or a combination of the two injuries in the same adult rat. The present study used three different fluorescent markers and a confocal laser image analysis system to study glial cells and phrenic motoneurons at the light microscopic level. Young adult female rats were divided into one combined injury group (left phrenicotomy and left C2 spinal hemisection with periods of 1 to 4 weeks between injuries, N = 12) and three other groups consisting of noninjured animals (N = 3), animals that received C2 hemisection only (N = 3), and animals with phrenicotomy only (survival periods of 2 (N = 3) and 4 (N = 3) weeks after phrenicotomy). Fluorogold was injected into the diaphragm to label phrenic motoneurons in all animals. Microglia and astrocytes were labeled with Texas red and fluorescein, respectively, and were visualized simultaneously along with phrenic motoneurons. The results suggest that the microglial and astrocytic response in the superimposed injury model are similar to the glial reactions characteristically seen in a peripheral axotomy alone model. These reactions include proliferation and migration of microglial cells along the perineuronal surface (peaking at 2 weeks) and the hypertrophy of astrocytes (peaking at 4 weeks). In addition, the increase in astrocytic tissue, which is characteristically seen in response to axotomy alone, is significantly enhanced in the superimposed injury model. Also, there is a large and rapid increase in GFAP-positive astrocytes within 24 hours after hemisection alone. The information gained from the present study will aid in determining, predicting, and eventually manipulating central nervous system responses to multiple injuries with the objective of reestablishing function in the damaged CNS. PMID- 9398445 TI - Presenilin 1 immunostaining using well-characterized antibodies in human tissues. AB - Using well-characterized monoclonal antibodies which recognize the N-terminus of presenilin 1 (PS1), we examined by immunohistochemistry brain tissues from patients with nonneurological conditions, Alzheimer's disease (AD) and cerebral infarction, as well as normal human liver tissues. The antibodies to PS1 did not reveal any positive staining in nonneurological conditions. In AD, the antibody showed positive staining of intraneuronal neurofibrillary tangles and neuropil threads. In cerebral infarcts, some macrophages were positively stained. In contrast to the C-terminal fragment of PS1, which has been claimed to be present in senile plaques, the N-terminal fragment binds to intracellular and intradendritic pathological structures and may play a role in tau phosphorylation in AD. PMID- 9398446 TI - Axotomized rubrospinal neurons rescued by fetal spinal cord transplants maintain axon collaterals to rostral CNS targets. AB - Neurons that maintain extensive axon collaterals proximal to the site of axotomy may be better able to survive injury. Early lesions of the rubrospinal tract lead to retrograde cell death of the majority of axotomized immature neurons. Transplants of fetal spinal cord tissue rescue axotomized rubrospinal neurons and promote their axonal regeneration. Rubrospinal neurons develop many of their axon collaterals postnatally. The present study tests the hypothesis that the axotomized rubrospinal neurons that are rescued by transplants and regenerate their axons are those neurons that have established axon collaterals to targets rostral to the lesion. Neonatal rats received a transplant of fetal spinal cord tissue placed into a midthoracic spinal cord hemisection. One month after transplantation, the retrogradely transported fluorescent tracers fast blue (FB) and diamidino yellow (DY) were used to identify rubrospinal neurons with collaterals to particular targets. FB was injected either into the interpositus nucleus of the cerebellum or into the gray matter of the cervical enlargement to identify collaterals to these targets, and DY was injected into the spinal cord approximately 5 mm caudal to the transplant and lesion site to label retrogradely the neurons that regenerated their axons. Double labeling was observed in the axotomized neurons of the red nucleus after tracer injections into the cervical spinal cord but not after injections into the cerebellum. This labeling pattern indicates that axotomized rubrospinal neurons that are rescued and regenerate axons caudal to the transplant maintain axon collaterals at cervical spinal cord levels. Cerebellar collaterals do not appear to play a role in the survival and regrowth of axotomized rubrospinal neurons. PMID- 9398447 TI - Neurotoxicity of endogenous cysteinylcatechols. AB - Progression of Parkinson's disease has been associated with several biochemical changes in the substantia nigra including increased oxidative challenge, catechol oxidation, and inhibition of mitochondrial complex I activity. Cysteinylcatechols, formed by nucleophilic addition of cysteine to oxidized catechols, have been identified as markers of catechol oxidation in brain tissue. We have examined the neurotoxicity of a series of cysteinylcatechols. Of the compounds examined, only 5-S-cysteinyl-3,4-dihydroxyphenylacetate (cysdopac) was specifically cytotoxic to differentiated P19 neuroglial cultures. Cysdopac also was neurotoxic to pyramidal neurons in organotypic cultures of hippocampus, and this effect was ablated by selective N-methyl-D-aspartate (NMDA) receptor antagonists. In vitro, cysdopac was a potent inhibitor of mitochondrial complex I activity. However, electrophysiologic experiments failed to demonstrate NMDA receptor agonist activity for cysdopac, nor did cysdopac inhibit glutamate uptake. These results showed that cysdopac was the most potent neurotoxin of this series of cysteinylcatechols and suggest that cysdopac may function as an indirect excitotoxin, potentially via inhibition of mitochondrial respiration. PMID- 9398448 TI - Immunolocalization of nitric oxide synthases at the postsynaptic domain of human and rat neuromuscular junctions--light and electron microscopic studies. AB - Neuronal (n) and inducible (i) nitric oxide synthase (NOS) were immunolocalized at the postsynaptic domain of human and rat neuromuscular junctions (NMJs) by light and electron microscopy. We applied polyclonal and monoclonal antibodies for colocalization with three other synaptic proteins, utilizing double and triple fluorescence labeling, and gold and peroxidase for immunoelectron microscopy. By light microscopy, nNOS and iNOS colocalized with desmin and dystrophin, known postsynaptic components, but not with neurofilament protein, a presynaptic component. By electronmicroscopy, nNOS, but not iNOS, colocalized postsynaptically on the same structures as desmin; iNOS was also postsynaptic, but did not colocalize with desmin immunoreactivity. At the NMJs of Duchenne muscular dystrophy patients, both nNOS and iNOS were strongly immunoreactive. At the NMJs of a patient with myasthenia gravis, nNOS was weaker than in controls. Total denervation of rat sciatic nerve did not cause any decrease of nNOS or iNOS immunoreactivity 7 days thereafter. At 15 days after denervation, there was a gradual decrease of immunoreactivity, and immunoreactivity disappeared 30 days after denervation, corresponding to the ultrastructurally detectable disorganization of the postsynaptic region. This seems to be the first combined light and electron microscopic description of the postsynaptic localization of nNOS and iNOS at human and rat NMJs. PMID- 9398449 TI - Effects of isradipine, an L-type calcium channel blocker on permanent and transient focal cerebral ischemia in spontaneously hypertensive rats. AB - Permanent or transient focal cerebral ischemia was induced in spontaneously hypertensive rats (SHR) using the intraluminal filament method. Successful occlusion of the middle cerebral artery (MCA) was achieved using 4/O filaments (terminal diameter 0.20-0.25 mm) coated with poly-L-lysine. The L-type calcium channel blocker isradipine (2.5 mg/kg) administered subcutaneously 30 min following permanent MCA occlusion significantly reduced the volume of ischemic brain damage in the cerebral hemisphere (25%; P = 0.0001), cerebral cortex (18%; P = 0.0034), and caudate nucleus (33%; P = 0.0002) when assessed at 24 h post-MCA occlusion. Isradipine did not affect the functional deficit (measured using a subjective neurological scoring system) induced by MCA occlusion. In SHR undergoing transient (2 h) MCA occlusion isradipine administered 30 min post-MCA occlusion produced a significant reduction (47%; P = 0.001) in hemispheric infarct volume, whereas isradipine administered at the onset of reperfusion did not confer any significant neuroprotection. No change in functional deficit was seen with isradipine with either dosing paradigm at 24 h post-MCA occlusion. These results demonstrate that the intraluminal filament method of MCA occlusion can be used in the SHR strain and also substantiates the neuroprotective efficacy of isradipine in SHR models of permanent and transient focal cerebral ischemia. PMID- 9398450 TI - Regenerating and sprouting axons differ in their requirements for growth after injury. AB - After spinal cord injury at birth, axotomized brainstem-spinal and corticospinal neurons are capable of permanent regenerative axonal growth into and through a fetal spinal cord transplant placed into the site of either a spinal cord hemisection or transection. In contrast, if fetal tissue which is not a normal target of the axotomized neurons (embryonic hippocampus or cortex) is placed into a neonatal spinal cord hemisection, brainstem-spinal serotonergic axons transiently innervate the transplant, but subsequently withdraw. The first set of experiments was designed to test the hypothesis that after spinal cord transection, serotonergic axons would cross the nontarget transplant, reach normal spinal cord targets caudal to the transection, and gain access to requisite target-derived cues, permitting permanent maintenance. Surprisingly, after a complete spinal cord transection, brainstem-spinal axons failed to grow into an inappropriate target even transiently. These observations suggest that the transient axonal ingrowth into nontarget transplants may represent lesion induced axonal sprouting by contralateral uninjured axons. We have used double labeling with fluorescent dyes, to test directly whether axonal sprouting of neurons which maintain collaterals to uninjured spinal cord targets (1) provide the transient ingrowth of brainstem-spinal axons into a nontarget transplant and (2) contribute to permanent ingrowth into target-specific transplants. Uninjured red nucleus, raphe nucleus, and locus coeruleus neurons extend axons into the nontarget transplant while maintaining collaterals to the host spinal cord caudal to the transplant. The lesion-induced sprouting by uninjured axons was also observed with a target-specific transplant. Taken together, these studies suggest that sprouting and regenerating axons may differ in their requirements for growth after injury. PMID- 9398451 TI - Increased expression of microtubule-associated protein 1B in the hippocampus, subiculum, and perforant path of rats treated with a high dose of pentylenetetrazole. AB - A single administration of the convulsant pentylenetetrazole (PTZ) initiates a complex pattern of long-term changes in microtubule-associated protein 1B (MAP1B) expression across the hippocampal formation. Using Northern blot and in situ hybridization we show that the first increases in MAP1B mRNA were detected at 15 h following PTZ administration in the granule cells of the dentate gyrus and CA3 region of the hippocampus and reached a maximum at 44 h. The levels of MAP1B mRNA in the subiculum peaked at later times (5 days). At 72 h MAP1B immunoreactivity was mainly localized in the granule-cell bodies and dentate inner and midmolecular layer as well as in neuronal cell bodies and the stratum lucidum, including the mossy fiber pathway of the CA3 region. By 5-10 days the levels of MAP1B in the pyramidal cells in the CA3 region decreased to very low levels; rather, heavy staining of interneuron-like cells and "strings-of-bead" structures all over the hippocampus and at the stratum oriens/alveus border were seen. The levels of MAP1B in the hippocampus returned to control levels by 20 days after PTZ administration. MAP1B immunoreactivity in the alvear path was also evident at 5 days postinjection at the CA1/alveus border. The intensity of MAP1B staining increased gradually in the perforant path starting at 72 h and persisted at high levels until day 35. Our studies show that (i) MAP1B is a temporal and regional marker for rapid and acute epileptic seizures and (ii) long-term increases in MAP1B in the perforant path might play a role in PTZ-induced seizures. PMID- 9398452 TI - Expression of brain-derived neurotrophic factor and TrkB neurotrophin receptors after striatal injury in the mouse. AB - Brain-derived neurotrophic factor (BDNF) promotes the survival and differentiation of nigral dopaminergic neurons and supports the activity of dopaminergic cells grafted into the striatum. However, little attention has been given to the physiological role of endogenous BDNF and its receptor TrkB within the nigrostriatal dopamine system. We know that striatal injury is followed by long-term stimulation of dopaminergic activity in the striatum, could BDNF play a role in this phenomenon? One week after physical injury to the striatum of C57/Black mice, just before dopaminergic activation becomes obvious, in situ hybridization on coronal sections through mouse striatum reveals that BDNF mRNA expression increases significantly before returning to basal levels within 1 month. Expression of mRNA for TrkB follows a very different pattern. No change of expression of the full-length and catalytically competent TrkBTK+ receptor is seen. However, expression of the truncated form of the receptor TrkTK-, which lacks the catalytic tyrosine kinase domain, does increase and stays elevated for at least 2 months after injury. When combined with observations of dopaminergic activation after striatal injury and the neuroprotective effects of BDNF introduced into the striatum, our findings suggest that BDNF and TrkBTK- do indeed play a role in dopaminergic regeneration and repair. PMID- 9398453 TI - Signals that regulate astroglial gene expression: induction of GFAP mRNA following seizures or injury is blocked by protein synthesis inhibitors. AB - Previous studies have revealed that a single electroconvulsive seizure (ECS) strongly induces glial fibrillary acidic protein (GFAP) expression in astrocytes in the hippocampal dentate gyrus. The signals that trigger this induction are not known, but circumstantial evidence suggests the hypothesis that GFAP expression may be induced as a result of the induction of growth factor expression by dentate granule cells that also occurs as a result of the ECS and other types of seizures. The present study tests one prediction of this hypothesis by evaluating whether increases in GFAP mRNA levels after ECS are blocked by inhibiting protein synthesis at various times after the ECS. We report that the upregulation of GFAP expression following ECS is blocked by protein synthesis inhibitors given 5 min before or up to 12 h after a single ECS. This temporal gradient suggests an intermediate step involving the increased expression of a protein growth factor. PMID- 9398454 TI - The attenuation of kainate-induced neurotoxicity by chlormethiazole and its enhancement by dizocilpine, muscimol, and adenosine receptor agonists. AB - Systemically administered kainate (10 mg.kg-1) caused neuronal loss in both the hippocampus and the entorhinal regions of the rat brain. This resulted in a loss of 68.3 +/- 13.8 and 53.3 +/- 12.8% of pyramidal neurones in the hippocampal CA1 and CA3a regions, respectively. Chlormethiazole attenuated the loss of neurones in the hippocampal cell layers CA1 (cell loss 10 +/- 3.2%) and CA3a (cell loss 10 +/- 7.7%). The neuroprotective activity of chlormethiazole was apparent in the presence or absence of a low dose of clonazepam (200 micrograms.kg-1 i.p.). The kainate-induced damage could also be measured by the increase in binding of the peripheral benzodiazepine ligand ([3H]PK11195) in the hippocampus. In kainate treated rats there was a 350-500% increase in binding indicative of reactive gliosis. Chlormethiazole prevented this elevation in a dose- and time-dependent manner, with an ED50 of 10.64 mg.kg-1 and an effective therapeutic window from 1 to 4 h posttreatment. Dizocilpine also attenuated damage significantly. The GABAA agonist muscimol was also able to attenuate the increase in [3H]PK11195 binding in a dose-dependent manner, with an ED50 of approximately 0.1 mg.kg-1. If muscimol, dizocilpine, or the adenosine A1 receptor agonist R-N6-phenylisopropyl adenosine were administered together with chlormethiazole at their respective ED25 doses, a potentiation was apparent in the degree of neuroprotection. It is concluded that the combination of neuroprotective agents with different mechanisms of action can lead to a synergistic protection against excitotoxicity. PMID- 9398455 TI - Triazolam impairs delayed recall but not acquisition of various everyday memory tasks in humans. AB - A double-blind test battery was administered to 24 human subjects (8 control, 16 drug) to assess the effects of 0.125 mg triazolam (oral) on memory encoding and retention across delay intervals ranging from seconds to 1 week after presentation. Although the drug reduced immediate psychomotor performance, it did not impair recall of previously learned information, nor did it significantly impair encoding of new information. The drug enhanced immediate recall of the location and identity of playing cards, without affecting 4-h delayed recall. The drug treatment impaired correct recall of object names after a delay of 20 min. At 4 h delay, the drug impaired olfactory recognition and free-recall of object names. At both 1 day and 1 week delay, the drug impaired recall of biographical information and correct identification of picture-photographer pair associations. The drug also impaired the daily improvement of the drug group as compared with the control group in a geometric puzzle solving task. The time course of these memory impairments compares well with the known effects of triazolam on long-term potentiation (LTP), a candidate biological mechanism underlying telencephalic memory formation and expression. PMID- 9398457 TI - In vitro cell density-dependent clonal growth of EGF-responsive murine neural progenitor cells under serum-free conditions. AB - Neural progenitor cell populations responsive to epidermal growth factor (EGF) have been shown to have proliferative potential and give rise to neurons, astrocytes, and oligodendrocytes. We have characterized EGF-responsive neural progenitor cells that give rise to bilineage neuronal/glial colonies (colony forming unit neuron-glia; CFU-NeGl) and unilineage neuronal colonies (CFU-Ne). Clonality was confirmed utilizing mixtures of brain cells from Balb/c and ROSA26 (transgenic for beta-galactosidase) mice. With a few exceptions, colonies showed either all blue cells or all clear cells after staining with X-Gal. Clonal growth was analyzed after 10-11 days in relation to cell density by determining colony size and plating efficiency. Growth was density dependent (no growth below 10,000 cell/ml) and thus single cell cloning was not accomplished. An average plating efficiency of 4% was found for EGF-responsive neural cells derived from day 15-18 murine embryos when cultured at 12,500 to 200,000 cells/ml. Similar results were obtained with 1-day-old postnatal neural cells. When colonies were categorized by size, the relative number of colonies over 50 cells appeared to be maximum at 50,000 plated cells/ml. After 11 days in culture, 94, 96, and 78% of the colonies contained cells that expressed nestin, neurofilament, and GFAP, respectively. Double-label experiments revealed that > 62% of the colonies contained both GFAP and neurofilament expressing cells. These studies establish the existence of at least two populations of clonal neural progenitors: CFU-Ne and CFU-NeGl in fetal and postnatal murine brain. PMID- 9398456 TI - Long-term survival of human central nervous system progenitor cells transplanted into a rat model of Parkinson's disease. AB - Progenitor cells were isolated from the developing human central nervous system (CNS), induced to divide using a combination of epidermal growth factor and fibroblast growth factor-2, and then transplanted into the striatum of adult rats with unilateral dopaminergic lesions. Large grafts were found at 2 weeks survival which contained many undifferentiated cells, some of which were migrating into the host striatum. However, by 20 weeks survival, only a thin strip of cells remained at the graft core while a large number of migrating astrocytes labeled with a human-specific antibody could be seen throughout the striatum. Fully differentiated graft-derived neurons, also labeled with a human-specific antibody, were seen close to the transplant site in some animals. A number of these neurons expressed tyrosine hydroxylase and were sufficient to partially ameliorate lesion-induced behavioral deficits in two animals. These results show that expanded populations of human CNS progenitor cells maintained in a proliferative state in culture can migrate and differentiate into both neurons and astrocytes following intracerebral grafting. As such these cells may have potential for development as an alternative source of tissue for neural transplantation in degenerative diseases. PMID- 9398459 TI - Adenovirus vector-mediated gene transfer into human epileptogenic brain slices: prospects for gene therapy in epilepsy. AB - As a first step in the development of a gene therapy approach to epilepsy, we evaluated the ability of adenovirus vectors to direct the transfer into and expression of a marker gene in human brain slices obtained from patients undergoing surgery for medically intractable epilepsy. Following injection of adenovirus vectors containing the Escherichia coli lacZ gene into hippocampal and cortical brain slices, lacZ mRNA, beta-galactosidase protein, and enzymatic activity were detected, confirming successful gene transfer, transcription, and translation into a functional protein. Transfected cells were predominantly glial, with some neurons expressing beta-galactosidase as well. These results support the potential of adenovirus vectors to transfer genetic information into human epileptogenic brain, resulting in expression of the gene into a functional protein. These findings also have implications for the development of gene therapy approaches to certain seizure disorders. A number of potential therapeutic approaches are discussed, including the elevation of inhibitory neurotransmitter or neuropeptide levels, expression or modulation of postsynaptic receptors, and manipulation of signal transduction systems. PMID- 9398458 TI - Neurotoxicity of polyamines and pharmacological neuroprotection in cultures of rat cerebellar granule cells. AB - We have studied in a well-characterized in vitro neuronal system, cultures of cerebellar granule cells, the toxicity of polyamines endogenously present in the brain: spermine, spermidine, and putrescine. Twenty-four-hour exposure of mature (8 days in vitro) cultures to 1-500 microM spermine resulted in a dose-dependent death of granule cells, with the half-maximal effect being reached below 50 microM concentration. Putrescine was moderately toxic but only at 500 microM concentration. Spermidine was tested at 50 and 100 microM concentration and its toxicity was evaluated to be about 50% that of spermine. Neuronal death caused by spermine occurred, at least in part, by apoptosis. Spermine toxicity was completely prevented by competitive (CGP 39551) and noncompetitive (MK-801) antagonists of the NMDA receptor, but was unaffected by a non-NMDA antagonist (NBQX) or by antagonists of the polyamine site present on the NMDA receptor complex, such as ifenprodil. A partial protection from spermine toxicity was obtained through the simultaneous presence of free radical scavengers or through inhibition of the free radical-generating enzyme nitric oxide synthase, known to be partially effective against direct glutamate toxicity. The link between spermine toxicity and glutamate was further strengthened by the fact that, under culture conditions in which glutamate toxicity was ineffective or much reduced, spermine toxicity was absent or very much decreased. Exposure to spermine was accompanied by a progressive accumulation of glutamate in the medium of granule cell cultures. This was attributed to glutamate leaking out from dying or dead cells and was substantially prevented by the simultaneous presence of MK-801 or CGP 39551. The present results demonstrate that polyamines are toxic to granule cells in culture and that this toxicity is mediated through the NMDA receptor by interaction of exogenously added polyamines with endogenous glutamate released by neurons in the medium. The involvement of brain polyamines, in particular spermine and spermidine, in excitotoxic neuronal death is strongly supported by our present results. PMID- 9398460 TI - Spinal Cord Transection-No Loss of Distal Ventral Horn Neurons AB - Anterograde transneuronal degeneration is caused by the loss of afferent input to the nerve cells and may occur in a number of neuronal systems. Transection of the adult spinal cord, causing anterograde transneuronal degeneration in ventral horn neurons, distal to the lesion, has been reported by some authors, while others contend that no such changes take place. The present study was undertaken in order to investigate whether transection of adult mouse thoracic spinal cord induces neuronal death in the ventral horns distal to the lesion. By means of modern stereological techniques such as the optical disector, the total number of cells in the lumbar ventral horns was estimated 7 weeks after transection. The mean numbers of neurons and glial and endothelial cells were 82,000 versus 89,000, 259,000 versus 301,000, and 129,000 versus 144,000 in the transected (n = 6) and sham-operated animals (n = 5), respectively. These differences were not statistically significant. Furthermore, neuronal soma volume was estimated by another stereological method, the vertical rotator. Mean neuronal soma volume was not significantly different between transected (2762 &mgr;m3) and sham-operated (2617 &mgr;m3) mice. Although no reduction in cell number or neuronal soma volume was observed, the mean volume of the ventral horns in the lumbar segments was significantly less in transected than in sham-operated animals, 2.49 mm3 versus 3.05 mm3 (P < 0.05). In conclusion, the transection of adult mouse thoracic spinal cord does not induce neuronal degeneration in the lumbar ventral horns. Copyright 1997 Academic Press. Copyright 1997 Academic Press PMID- 9398461 TI - Generation and transplantation of EGF-responsive neural stem cells derived from GFAP-hNGF transgenic mice. AB - EGF-responsive neural stem cells isolated from murine striatum have the capacity to differentiate into both neurons and glia in vitro. Genetic modification of these cells is hindered by a number of problems such as gene stability and transfection efficiency. To circumvent these problems we generated transgenic mice in which the human GFAP promoter directs the expression of human NGF. Neural stem cells isolated from the forebrain of these transgenic animals proliferate and form clusters, which appear identical to stem cells generated from control animals. Upon differentiation in vitro, the transgenic stem cell-derived astrocytes express and secrete bioactive hNGF. Undifferentiated GFAP-hNGF or control stem cells were transplanted into the striatum of adult rats. One and 3 weeks after transplantation, hNGF was detected immunocytochemically in an halo around the transplant sites. In GFAP-hNGF-grafted animals, intrinsic striatal neurons proximal to the graft appear to have taken up hNGF secreted by the grafted cells. Ipsilateral to implants of GFAP-hNGF-secreting cells, choline acetyltransferase-immunoreactive neurons within the striatum were hypertrophied relative to the contralateral side or control-grafted animals. Further, GFAP-hNGF grafted rats displayed a robust sprouting of p75 neurotrophin receptor-positive fibers emanating from the underlying basal forebrain. These studies indicate that EGF-responsive stem cells which secrete hNGF under the direction of the GFAP promoter display in vitro and in vivo properties similar to that seen following other methods of NGF delivery and this source of cells may provide an excellent avenue for delivery of neurotrophins such as NGF to the central nervous system. PMID- 9398462 TI - Serotonin promotes the survival of cortical glutamatergic neurons in vitro. AB - The appearance of 5-hydroxytryptamine (serotonin; 5-HT) in the cerebral cortex coincides with developmental events such as cell proliferation, survival, and differentiation. We tested the hypothesis that 5-HT plays a role in these events by examining rat cortical progenitor cells in vitro. Using bromodeoxyuridine incorporation we found that 5-HT did not affect the proliferation of these cells, but a cell survival assay indicated that it promoted their survival. The observed survival effect was mimicked by the 5-HT2a/2c receptor agonist alpha-methyl-5-HT and blocked by the 5-HT2a receptor antagonist cinanserin. Consistent with increased survival was the finding, using the terminal transferase nick end labeling method, of reduced cell death in cultures exposed to 5-HT. Immunohistochemical analysis with cell-specific markers revealed that the effect of 5-HT was directed specifically to the glutamate-containing neuronal population and not to any other cortical cell types. These results indicate that 5-HT does not exert its effects on dividing neuroepithelial cells in the developing cortex, but rather on postmitotic neurons. PMID- 9398463 TI - Hydrocephalus in the Otx2+/- mutant mouse. AB - Mice with the Otx2+/- mutation often die during the postneonatal period. Before death these animals, generated from TT2 ES cells and crossed with CBA mice, develop a dome-shaped head, weakness of the limbs, kyphosis, lethargy, drowsiness, and emaciation. Autopsy of these mice revealed eminent dilatation of lateral ventricles and a ballooned cerebrum. Histological analysis shows edematous change of the periventricular white matter. These results suggest that Otx2 functions as a head organizer, and a mutation of this gene is a likely cause of hydrocephalus in mammals. Additionally, craniobasal skeletal anomaly in half of the heterozygotes and dwarfism in some of the female heterozygotes are described. PMID- 9398464 TI - Regulation of D-aspartate release and uptake in adult brain stem auditory nuclei after unilateral middle ear ossicle removal and cochlear ablation. AB - In young adult guinea pigs, the effects of unilateral ossicle removal and cochlear ablation were determined on transmitter release from glutamatergic presynaptic endings and glutamate inactivation via uptake. (i) D-[3H]Aspartate release and uptake were measured in subdivisions of the cochlear nucleus (CN) and in nuclei of the superior olive (SOC) and auditory midbrain (MB) up to 145 days after placing the lesions. Activities were compared to those from age-matched unlesioned controls. Fiber degeneration was visualized histologically. (ii) In the ipsilateral CN, changes in release and uptake were governed by the type of lesion. Ossicle removal produced sparse pruning of fibers only after 112 days and decreased release and uptake at 145 days, consistent with regulatory weakening of excitatory glutamatergic transmission. Cochlear ablation deafferented the CN, producing deficient release and uptake at 2 days and abundant fiber degeneration at 7 days. Subsequently, the residual release and uptake increased in magnitude, consistent with strengthening of excitatory glutamatergic transmission. (iii) In the contralateral CN, after either lesion, changes in release and uptake usually matched those in the ipsilateral CN. Thus, the auditory pathway associated with the lesioned ear probably provided cues for the regulation of synaptic strength in the contralateral CN. (iv) Both lesions increased release in the SOC and MB, and uptake in the SOC, consistent with strengthening of excitatory glutamatergic transmission. Sparse fiber degeneration, suggesting axonal pruning, appeared in the SOC and MB after cochlear ablation. (v) The strengthening of excitatory glutamatergic transmission may facilitate and maintain symptoms such as loudness recruitment and tinnitus which often accompany hearing loss. PMID- 9398465 TI - Sciatic nerve regeneration through venous or nervous grafts in the rat. AB - This study analyses the interest of isologous venous grafts filled with saline or with Schwann cells versus nerve grafts as guides for regeneration of the sciatic nerve in 35 Wistar rats. Electrophysiological parameters (conduction velocities and distal latencies of motor responses) and the functional index of De Medinacelli were measured several times from 1 month to 1 year after surgery. An histological analysis was performed on 2 control rats and on 3 rats killed 6 or 12 months after surgery: the total number of fibers was counted on a montage photoprint of the whole nerve, and the diameters of axons and the thickness of the myelin sheath were measured on digitized images. With a portion of nerve as guide, the regeneration is faster than with a vein. However, regeneration after 6 months is at least as good with a venous graft filled with Schwann cells, as assessed by electrophysiological, functional, and histological analysis. The addition of Schwann cells in grafted veins allows the nerve to regenerate through longer gaps than previously described (25 vs 15 mm). In order to assess the quality of nerve regeneration, functional, electrophysiological, and histological analysis are complementary. PMID- 9398466 TI - Genetic transfer of the wobbler gene to a C57BL/6J x NZB hybrid stock: natural history of the motor neuron disease and response to CNTF and BDNF cotreatment. AB - Preclinical diagnosis of motor neuron disease (MND) in the wobbler mouse (wr/wr) has been impossible until recently. However, with the development of a new hybrid, the C57BL/6J x New Zealand Black (B6NZB) wr/wr mouse, the polymerase chain reaction (PCR) can be used to establish the preclinical diagnosis. We compared the clinical and histological features of MND and the effects of neurotrophic factor cotreatment between the hybrid B6NZB-wr/wr and the congenic C57BL/6J-wr/wr mice. Clinical assessments of body weight, grip strength, running speed, paw position, and walking pattern were made weekly from age 2 weeks through 8 weeks (n = 10, B6NZB-wr/wr; n = 15, C57BL/6J-wr/wr). Survival was analyzed (n = 7, each strain) as was C5 and C6 spinal cord motoneuron morphology and ventral root histometry (n = 7, each strain). For cotreatment, 8 B6NZB-wr/wr and 7 C57BL/6J-wr/wr mice received subcutaneous ciliary neurotrophic factor (1 mg/kg) and brain-derived neurotrophic factor (5 mg/kg) on alternate days, 6 days/week for 4 weeks. B6NZB-wr/wr mice could be distinguished from C57BL/6J wr/wr mice at age 3 weeks by a more abnormal paw position (P < 0.01) and walking pattern (P < 0.05) and lower grip strength (P < 0.001) and running speed (P < 0.001). After 3 weeks, the changes continued to be greater in B6NZB-wr/wr mice. Although B6NZB-wr/wr mice were more severely affected early in the disease, their survival was comparable to C57BL/6J-wr/wr mice. Anterior horn cell vacuolar degeneration and myelinated fiber histometry were similar in both strains. The clinical response to CNTF/BDNF cotreatment was marked in both groups although it was weaker in B6NZB-wr/wr mice. Thus, the hybrid B6NZB-wr/wr mice have a more severe clinical phenotype and offer a unique opportunity to study the mechanisms of presymptomatic motor neuron degeneration and the effects of therapeutic agents for human MND. PMID- 9398467 TI - Magnetic resonance imaging and behavioral analysis of immature rats with kaolin induced hydrocephalus: pre- and postshunting observations. AB - The motor and cognitive dysfunction associated with hydrocephalus remains a clinical problem in children. We hypothesized that young rats with hydrocephalus should exhibit similar dysfunction and that the dysfunction should be reversible by shunting. Hydrocephalus was induced in 3-week-old rats by injection of kaolin into the cisterna magna. Rats were assessed by T2-weighted images obtained with a 7-T magnetic resonance device and by repeated behavioral testing including ability to traverse a narrow beam and ability to find a hidden platform in a water pool. Some of the rats underwent a shunting procedure 1 or 4 weeks after kaolin injection. Magnetic resonance images were used to measure ventricle size. They clearly demonstrated increased signal in periventricular white matter, which corresponded to increased brain water content. A flow-void phenomenon was observed in the cerebral aqueduct. Ability to traverse the beam did not correlate with the degree of ventriculomegaly. Ability to swim to the hidden platform demonstrated a progressive impairment of learning function which may have been accentuated by motor disability. When rats were shunted after 1 week, the behavioral dysfunction was prevented. Late shunting after 4 weeks was associated with gradual recovery of the behavioral disability which was not complete after 4 weeks. We conclude that early shunting is superior to late shunting with regard to behavioral dysfunction. High-resolution MR imaging shows features in hydrocephalic rats similar to those found in hydrocephalic humans. PMID- 9398469 TI - Effect of embryonic donor age and dissection on the DARPP-32 content of cell suspensions used for intrastriatal transplantation. AB - The aim of this study was to determine in vitro the DARPP-32 content of donor cells used for striatal transplantation in vivo. The effect of selective embryonic dissection of the lateral ganglionic eminence (LGE) was compared with the standard dissection of the whole ganglionic eminence (WGE) at each of three embryonic ages (14, 15, and 16 days of gestation) in the rat. The resultant cell suspensions were cultured for up to 7 days and incubated with antibodies against DARPP-32, a marker of striatal medium spiny neurons; beta-tubulin III, a neuronal marker; GFAP, a marker of reactive astrocytes; and Gal-C, a marker of oligodendrocytes. LGE dissection gave rise to more DARPP-32 neurons compared to WGE; but this relationship was only observed in the younger embryos. When older (16 days gestation) embryos are used there is no difference in the yield of DARPP 32 cells obtained from LGE and WGE. LGE dissections were also observed to contain fewer glial cells. There was no beneficial effect of LGE over WGE on survival of striatal neurons in vitro. These results have important implications for the selection and dissection of fetal donor material used in clinical trials of intrastriatal transplantation as a potential treatment for Huntington's disease. PMID- 9398470 TI - Densitometric quantification of neuronal viability by computerized image analysis. AB - A new method is presented for the quantification of cell viability based on densitometry with computerized image analysis. Neuronal cells were stained with crystal violet and densitometric analysis was performed with an IBAS 2.0 image analyzer (Kontron/ Zeiss), using specially implemented dedicated software which integrates the optical density of the culture in each well with the area covered by the stained cells. To test the reliability of the densitometric method cortical cells were plated at different concentrations (5 x 10(4)-10(6)/ml); the standard curve obtained by analysis of crystal violet staining showed a linear proportion between cell number and optical density signal. The validation and accuracy of the method were assessed and compared with other methods using rat cortical cells cultured in vitro for 10 days and exposed to kainic acid (250 microM) for 24 h. Neuronal viability was reduced by 40-50% and comparison with direct cell counting, MTT assay, and spectrophotometric analysis confirmed that the method is simple, quick, and reliable. PMID- 9398468 TI - The cannabinoid receptor agonist WIN 55,212-2 reduces D2, but not D1, dopamine receptor-mediated alleviation of akinesia in the reserpine-treated rat model of Parkinson's disease. AB - The effects of the synthetic cannabinoid receptor agonist WIN 55,212-2 on dopamine receptor-mediated alleviation of akinesia were evaluated in the reserpine-treated rat model of parkinsonism. The dopamine D2 receptor agonist quinpirole (0.1 mg/kg, ip) caused a significant alleviation of the akinesia. This effect was significantly reduced by coinjection with the cannabinoid receptor agonist WIN 55,212-2 (0.1 and 0.3 mg/kg). The simultaneous administration of the cannabinoid receptor antagonist SR 141716A (3 mg/kg, ip) with quinpirole and WIN 55,212-2 blocked the effect of WIN 55,212-2 on quinpirole-induced alleviation of akinesia. The selective dopamine D1 receptor agonist chloro-APB (SKF82958, 0.1 mg/kg) alleviated akinesia in a significant manner. WIN 55,212-2 (0.1-1 mg/kg, ip) did not affect the antiakinetic effect of chloro-APB. Combined injection of both D1 and D2 dopamine receptor agonists (both at either 0.1 or 0.02 mg/kg) resulted in a marked synergism of the antiakinetic effect. WIN 55,212-2 (0.1-1 mg/kg) significantly reduced the antiakinetic effect of combined injections of quinpirole and chloro-APB at both 0.1 and 0.02 mg/kg. The effect of 0.3 mg/kg WIN 55,212-2 on combined D1 and D2 agonist-induced locomotion (0.02 mg/kg) was blocked by SR 141761A (3 mg/kg). Neither WIN 55,212-2 alone (0.1 and 0.3 mg/kg) nor SR 141716A (3 and 30 mg/kg) alone had an antiparkinsonian effect. These results suggest that cannabinoids may modulate neurotransmission in the pathway linking the striatum indirectly to basal ganglia outputs via the lateral globus pallidus and the subthalamic nucleus. PMID- 9398471 TI - Effects of different schedules of MPTP administration on dopaminergic neurodegeneration in mice. AB - Although a valuable 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) animal model of human Parkinson's disease has been developed, our knowledge of the course of nigral degeneration remains fragmentary. Experimental factors which could possibly influence the destructive process must be taken into account. To evaluate the impact of experimental design, we compared the effects of different schedules of injection of the same cumulative dose of MPTP, in mice, by measuring tyrosine hydroxylase immunoreactivity in the substantia nigra pars compacta. Massive injection of the total dose over 1 day (4 injections of 20 mg/kg) destroyed more dopaminergic neurons than did the long-term daily injections of a lower dose of MPTP (20 injections of 4 mg/kg). This suggests that different schedules of administration of MPTP might induce different mechanisms of neuronal death. These mechanisms need to be better understood if chronic models of intoxication that replicate the evolution of human Parkinson's disease more precisely are to be developed. PMID- 9398472 TI - Chronic hypoxia does not induce synaptic plasticity in the phrenic nucleus. AB - Interruption of the main descending respiratory drive to phrenic motoneurons by cold block or spinal cord hemisection results in morphological modifications of the ipsilateral phrenic nucleus in the rat. The modifications consist of an increase in the number of multiple synapses and dendrodendritic appositions and elongation of the asymmetric and symmetric synaptic active zones. Hemisection and hemispinalization by cold block cause not only "functional deafferentation" of the ipsilateral phrenic neurons (i.e., a loss of ipsilateral descending respiratory drive), but also an increase in the remaining contralateral descending respiratory drive. The contralateral respiratory pathways connect with phrenic motoneurons ipsilateral to cold block or hemisection by decussating collateral axons which cross the spinal cord midline below the hemisection/cold block site. Thus, the phrenic nucleus synaptic plasticity could possibly be induced by functional deafferentation or by an increase of the descending respiratory drive. To differentiate between these two possible inducers of the plasticity, we assessed the synaptic morphology of the phrenic nucleus of nonoperated rats exposed to 48 h of hypoxia in an atmosphere chamber. The hypoxia exposure produces an increased descending respiratory drive without functional deafferentation. The quantitative data extracted from electron micrographs of the phrenic nucleus from four experimental rats were compared with the data from four normal breathing animals. Phrenic nucleus morphometric analysis showed that there was no significant difference in the mean number of single synapses between the samples from control animals (141 +/- 12.12) and the experimental animals (156 +/ 26.73). Similarly, no significant difference was detected in the total number of synaptic active zones of control animals (178.25 +/- 11.13) and experimental animals (195.05 +/- 5.35). Furthermore, the length of synaptic active zones of asymmetrical synapses (0.21 +/- 0.024 micron) or symmetrical synapses (0.22 +/- 0.022 micron) did not change significantly compared to the synaptic active zone length in control animals (0.21 +/- 0.018 micron for asymmetrical and 0.21 +/- 0.010 micron for symmetrical). We conclude that no synaptic plasticity occurs in the phrenic nucleus without functional deafferentation in spite of an increase in descending respiratory drive. Therefore functional deafferentation may be the primary inducer of phrenic nucleus synaptic plasticity occurring after hemisection or cold block. PMID- 9398473 TI - Delayed maturation of regenerating myelinated axons in mice lacking neurofilaments. AB - Using the technique of homologous recombination in embryonic stem cells, we generated mice bearing a targeted disruption of the gene encoding the neurofilament light (NF-L) protein. The absence of NF-L protein in mice resulted in dramatic declines of approximately 20-fold in the levels of neurofilament medium and heavy proteins in the brain and sciatic nerve while increases were detected for other cytoskeletal proteins such as tubulin and GAP-43. Despite a lack of neurofilaments and hypotrophy of axons, the NF-L knockout mice develop normally and do not exhibit overt phenotypes. However, in both NF-L -/- and NF-L +/- mice, the regeneration of myelinated axons following crush injury of peripheral nerves was found to be abnormal. In the second week after axotomy, the number of newly regenerated myelinated axons in the sciatic nerve and facial nerve of NF-L -/- mice corresponded to only approximately 25 and approximately 5% of the number of myelinated axons found in normal mice, respectively. At this early postaxotomy stage, electron microscopy of nerve segments distal to the crush site in NF-L -/- mice revealed abundant clusters of axonal sprouts that were indicative of retarded maturation of regenerating fibers. The analysis of the distal sciatic nerve at 2 months after crush indicated that neurofilament deficient axons have the capacity to regrow for a long distance and to remyelinate, albeit at a slower rate. These results provide the first direct evidence for a role of neurofilaments in the maturation of regenerating myelinated axons. PMID- 9398474 TI - Effects of prenatal protein malnutrition on hippocampal long-term potentiation in freely moving rats. AB - It has been demonstrated that prenatal protein malnutrition significantly affects hippocampal plasticity, as measured by long-term potentiation, throughout development. This paper focuses on the hippocampal dentate granule cell population response to two separate paradigms of tetanization of the medial perforant pathway in prenatally protein-malnourished and normally nourished adult male rats. The 100-pulse paradigm consisted of the application of ten 25-ms duration bursts of 400 Hz stimulation with an interburst interval of 10 s. The 1000-pulse paradigm consisted of the application of five 500-ms bursts of 400 Hz stimulation with an interburst interval of 5 s. No between-group differences were obtained for input/output response measures prior to tetanization. No between group, nor between-paradigm, differences were obtained in the degree of population EPSP slope enhancement. However, in response to both paradigms, prenatally malnourished animals showed significantly less enhancement of the population spike amplitude (PSA) measure than normally nourished animals. Normally nourished animals showed a significantly greater level of PSA enhancement in response to the 100-pulse paradigm than the 1000-pulse paradigm. Prenatally malnourished animals showed no significant differences in the degree of PSA enhancement between the two paradigms. Results indicate that short duration bursts (< or = 25 ms) are more effective in inducing maximal PSA enhancement in normally nourished rats than longer duration stimulus bursts. The apparent inability of prenatally malnourished rats to transfer enhanced cellular activation (population EPSP slope enhancement) into enhanced cellular discharge (PSA enhancement) suggests that a preferential enhancement of GABAergic inhibitory modulation of granule cell excitability may result from the prenatal dietary insult. Such potentiation of inhibitory activity would significantly lower the probability of granule cell population discharge, resulting in the significantly lower level of PSA enhancement obtained from these animals. PMID- 9398475 TI - Tirilazad mesylate increases dopaminergic neuronal survival in the in Oculo grafting model. AB - Grafting of fetal ventral mesencephalon in Parkinson's disease has been extensively studied. A crucial draw back of this technique is the low survival rate of the dopaminergic neurons. It has been documented that only 5-20% of the grafted neurons survive, and to enhance graft efficacy to a satisfying level, increased cell survival is of utmost desire. In this study we have used the antioxidant tiriliazad mesylate (U-74006F) to study the effect on the survival of dopaminergic neurons after grafting. The in oculo grafting model was used and ventral mesencephalon was dissected from E14-E15 rat fetuses in Hanks' balanced salt solution (HBSS), in Dulbecco's modified Eagle medium (DMEM), or in 0.3, 3.0, or 30 microM U-74006F diluted in DMEM. The tissue was then inserted into the anterior chamber of the eye. Some of the transplants were further treated with intraocular injections of 3 or 30 microM U-74006F (5 microliters) weekly for 2 weeks. Quantification of tyrosine hydroxylase (TH)-immunoreactive profiles revealed that in transplants treated with U-74006F at dissection only, no change in the number of TH-positive neurons was found. Pretreatment of 0.3 microM U 74006F during dissection combined with intraocular injections of U-74006F after grafting, on the other hand, resulted in a dose-dependent enhancement of survival of TH-positive neurons. Dissection in, and intraocular treatment with, 3 microM U 74006F resulted in a significantly enhanced survival of TH-positive neurons whereas using U-74006F at a concentration of 30 microM did not change the cell survival compared to solely DMEM-treated grafts. Thus, 30 microM was interpreted to be an overdose. Comparing cell survival when dissected in DMEM with that dissected in HBSS showed that DMEM was clearly superior. Nerve fiber formation was most pronounced in grafts treated with 3 microM U-74006F. In conclusion, survival of TH-positive neurons is enhanced by U-74006F, which is readily available for clinical use and thus could be employed to enhance graft survival when transplanting patients suffering from Parkinson's disease. PMID- 9398476 TI - Rat intrastriatal neural allografts challenged with skin allografts at different time points. AB - The present study was designed to address two questions. First, can an intrastriatal neural allograft exhibit long-term survival (18 weeks) if the host is immunized by an orthotopic skin graft 6 weeks after neural transplantation (the 6w-Long group)? Second, can an intrastriatal neural allograft survive when the host is challenged by an orthotopic skin allograft either simultaneously (Sim) with the intracerebral graft surgery or 2 (2w) weeks later? Dissociated embryonic ventral mesencephalic tissue from Lewis rats was stereotaxically injected into the striatum of Sprague-Dawley rats with unilateral 6 hydroxydopamine lesions. Six weeks after neural grafting, no reduction in amphetamine-induced motor asymmetry was observed in the Sim and 2w groups. At 6 weeks after skin grafting, the mean motor asymmetry scores had returned to the initial pretransplantation levels in the 6w-Long group. All the neural allografts in the Sim group were completely rejected, and the mean number of tyrosine hydroxylase immunoreactivity neurons in the grafts was significantly reduced in the 2w and the 6w-Long group, when compared to the no-skin control group. There were very high levels of expression of MHC class I and II antigens, marked cellular infiltrates containing macrophages and T-lymphocytes, and several activated microglia and astrocytes in and around the surviving intracerebral transplants in the 2w and the 6w-Long groups. The results suggest that intrastriatal neural allografts are more likely to be rejected rapidly if the host is efficiently immunized with the same alloantigens simultaneously or soon after the neural transplantation than at a later time point. When established neural allografts are subjected to a strong immunological challenge, they undergo protracted rejection. PMID- 9398477 TI - Regulation of astroglial-derived dopaminergic neurotrophic factors by interleukin 1 beta in the striatum of young and middle-aged mice. AB - Interleukin-1 beta (IL-1 beta) can induce dopaminergic axonal sprouting in the denervated striatum of parkinsonian animals. In order to determine whether IL-1 beta effects on dopaminergic axonal sprouting are mediated by the induction of astroglial-derived dopaminergic neurotrophic factors, effects of IL-1 beta treatment on acidic and basic fibroblast growth factor (aFGF and bFGF) and glial cell line-derived growth factor (GDNF) gene expression were examined in primary striatal astrocyte cultures and after in vivo administration. We found a selective induction of bFGF mRNA synthesis but not aFGF or GDNF mRNA after IL-1 beta treatment both in vitro and in vivo. This suggests that bFGF may be the putative endogenous dopaminergic neurotrophic factor mediating lesion-induced plasticity of dopamine neurons. In addition, to determine why recovery from injury becomes reduced with age, we examined whether there was an aging associated decline in the ability of IL-1 beta to induce the synthesis of neurotrophic factors in middle-aged animals compared to young mice. Interestingly, IL-1 beta stimulated a greater induction in bFGF mRNA levels in the middle-aged mice compared to young mice. These results suggest that the regulation of bFGF and possibly its receptor signaling efficacy may vary as the brain ages. PMID- 9398478 TI - The effects of variable periods of functional deprivation on olfactory bulb development in rats. AB - Dramatic alterations occur in the developing olfactory bulb when air flow is reduced through one-half of the nasal cavity. Naris closure on the day after the day of birth (P1) in rats, for example, results in reduced cell survival in the ipsilateral bulb by P20 and a substantial (25%) decrease in bulb size by P30. Almost immediate changes in protein synthesis and cell metabolism are also observed, and one prevalent theory suggests that these changes may be important in specifying which cells are subsequently eliminated. In the present study we used a reversible technique for unilateral naris closure to examine the sensitive period for the effects of olfactory deprivation on bulb size and cell survival. This technique involves the insertion of removable plugs into a rat pup's external naris. We occluded the naris for increasing periods of time (P1-P10, P1 P15, or P1-P20), reared all animals to P30, and measured volumes of bulb laminae. In addition, we examined the duration of naris closure needed to affect cell survival by injecting animals with the thymidine analogue bromodeoxyuridine to label cells born soon after the onset of olfactory deprivation. Results indicate that relatively long periods of naris occlusion (P1-P15 or longer) are required to produce a substantial reduction in experimental bulb size. Cell survival was decreased following olfactory deprivation from P1 to P10, but not after deprivation from P1 to P3. These data support the hypothesis that changes that occur within 48 h of naris closure are not sufficient to affect cell survival. PMID- 9398479 TI - c-Jun expression in adult rat dorsal root ganglion neurons: differential response after central or peripheral axotomy. AB - The response of the mature central nervous system (CNS) to injury differs significantly from the response of the peripheral nervous system (PNS). Axotomized PNS neurons generally regenerate following injury, while CNS neurons do not. The mechanisms that are responsible for these differences are not completely known, but both intrinsic neuronal and extrinsic environmental influences are likely to contribute to regenerative success or failure. One intrinsic factor that may contribute to successful axonal regeneration is the induction of specific genes in the injured neurons. In the present study, we have evaluated the hypothesis that expression of the immediate early gene c-jun is involved in a successful regenerative response. We have compared c-Jun expression in dorsal root ganglion (DRG) neurons following central or peripheral axotomy. We prepared animals that received either a sciatic nerve (peripheral) lesion or a dorsal rhizotomy in combination with spinal cord hemisection (central lesion). In a third group of animals, several dorsal roots were placed into the hemisection site along with a fetal spinal cord transplant. This intervention has been demonstrated to promote regrowth of severed axons and provides a model to examine DRG neurons during regenerative growth after central lesion. Our results indicated that c-Jun was upregulated substantially in DRG neurons following a peripheral axotomy, but following a central axotomy, only 18% of the neurons expressed c-Jun. Following dorsal rhizotomy and transplantation, however, c-Jun expression was upregulated dramatically; under those experimental conditions, 63% of the DRG neurons were c-Jun-positive. These data indicate that c-Jun expression may be related to successful regenerative growth following both PNS and CNS lesions. PMID- 9398480 TI - Predegenerated nerve allografts versus fresh nerve allografts in nerve repair. AB - This study reevaluated the possibility of using predegenerated nerves as donor nerve allografts for nerve repair and compared the results of functional recovery to those obtained after standard, fresh nerve allograft repair. Twenty donor rats underwent a ligature/ section of the left sciatic nerve 4 weeks before nerve graft harvesting. Forty recipient rats underwent severing of the left sciatic nerve leaving a 15-mm gap between the nerve stumps. Graft repair was undertaken using either the predegenerated left sciatic nerve of the 20 donor rats (predegenerated group, 20 recipient rats) or the normal right sciatic nerve of the 20 donor rats (fresh group, 20 recipient rats). Recovery of function was assessed by gait analysis, electrophysiologic testing and histologic studies. Walking tracks measurements at 2 and 3 months, electromyography parameters at 2 and 3 months, peroperative nerve conduction velocity and nerve action potential amplitude measurements at 3 months, as well as assessments of myelinated nerve fiber density and surface of myelination showed that fresh and predegenerated nerve grafts induced a comparable return of function although there was some trend in higher electrophysiologic values in the predegenerated group. The only slight but significant difference was a larger mean nerve fiber diameter in the nerve segment distal to a predegenerated nerve graft compared to a fresh nerve graft. Although our study does not show a dramatic long-term advantage for predegenerated nerve grafts compared to fresh nerve grafts, their use as prosthetic material is encouraging. PMID- 9398481 TI - Intracerebral transplantation of testis-derived sertoli cells promotes functional recovery in female rats with 6-hydroxydopamine-induced hemiparkinsonism. AB - Recently, we demonstrated amelioration of behavioral deficits associated with 6 hydroxydopamine-induced hemiparkinsonism by transplanting rat testis-derived Sertoli cells into adult male rat brains. In the present study, we used adult female hemiparkinsonian rats to investigate whether the beneficial effects of transplantation of Sertoli cells may be differentially affected by gender of the animal transplant recipient. At 1 month posttransplantation, animals transplanted with Sertoli cells showed functional recovery as revealed by significant reductions in apomorphine-induced rotational behavior and asymmetrical elevated body swing behavior. Control animals that received medium alone did not display any visible behavioral recovery. These results suggest that transplantation of Sertoli cells is not male hormone-dependent and further support the use of these cells as a graft source for Parkinson's disease and other neurological disorders. PMID- 9398482 TI - LETTER TO THE EDITOR PMID- 9398483 TI - Genetic polymorphisms in human drug metabolic enzymes. AB - Results obtained from both epidemiologic studies and experimental animal model systems have shown a wide range of phenotypic variation in the ability of individuals to metabolize drugs and environmental chemicals. Several studies have noted correlations between specific metabolic phenotypes and the incidence of disease, suggesting that certain allelic forms of drug metabolic enzymes can render the individual either more sensitive or resistant to the toxic or therapeutic effects of exogenous drugs and chemicals. While some of this variation can be attributed to different environmental exposures, it has become clear that genetic factors also play an important role in determining the response of the individual organism to exogenous agents. Recent advances in molecular biological techniques have begun to allow scientists to correlate observed phenotypic differences with the actual differences in genetic sequence at the gene level. This has allowed a correlation between gene structure and function, thus providing a mechanistic basis to explain the interaction between genetic background and individual response to environmental exposures. Results presented at this symposium discussed how genetic polymorphisms for both Phase I and Phase II metabolic enzymes in the human population modulate the response to environmental toxicants. PMID- 9398484 TI - Subchronic nasal toxicity of hexamethylphosphoramide administered to rats orally for 90 days. AB - Rats were administered hexamethylphosphoramide (HMPA) at dosages of 10, 100, 300, and 1000 ppm in drinking water or at 15, 40, or 120 mg/kg/day by gavage for approximately 90 days. Another group of rats was implanted subcutaneously with HMPA-filled osmotic minipumps, designed to deliver a dosage of 40 mg/kg/day to prevent the possibility of direct contact of HMPA with the nasal epithelium. After 90 days at 10 ppm in the drinking water, some rats had tracheas lined with regenerated epithelium, but no HMPA-related lesions were present in any other organs and tissues. At 100 ppm, nasal lesions (epithelial denudation, regeneration, and squamous metaplasia) were mostly in the maxilloturbinates, tips of nasoturbinates, and the adjacent septum in the anterior nasal cavity (level I), but the lesions were confined to the ventral region of the mid-anterior nasal cavity (level II) and to recesses of the posterior nasal cavity (levels III and IV). At 300 ppm, nasal turbinates in level I were partially adhered to the nasal septum by fibrous tissue. In level II the lesions were mainly confined to the ventral medial meatus, but were scattered diffusely in levels III and IV. Denuded turbinates showed minimal bone proliferation. At 1000 ppm, the anterior nasal cavity was partially occluded by extensive adhesion of the turbinates to the nasal septum by granulation tissue and proliferating turbinate bone. The general architecture of the posterior nasal cavity was obliterated by the marked proliferation of turbinate bone and fibrous tissue in the interturbinate spaces. Tracheas showed regenerated epithelium and bronchi had focal epithelial denudation at 100, 300, and 1000 ppm. Foamy alveolar macrophages (histiocytosis) were increased in the lungs at 300 and 1000 ppm. Testicular atrophy occurred at 1000 ppm. No other tissues were affected by HMPA treatment. Nasal lesions in rats given HMPA by gavage were identical in nature to, but sometimes slightly more severe than, the lesions in rats given HMPA in the drinking water. Rats given 40 mg/kg/day HMPA via an osmotic minipump had slightly less severe nasal lesions than did the rats given the same dosage of HMPA by gavage. Testicular atrophy was present in the rats given 120 mg/kg/day by gavage. The results of this study show that, with the exception of bone proliferation, systemic delivery of HMPA or its metabolites to the nasal tissue following oral administration causes tissue damage similar to that caused by direct exposure of the nasal tissue via inhalation. Oral administration of HMPA is a less potent route for producing nasal lesions than is inhalation. PMID- 9398485 TI - Effect of dosing vehicle on the developmental toxicity of bromodichloromethane and carbon tetrachloride in rats. AB - Several halocarbons have been shown to cause full-litter resorption (FLR) in Fischer-344 rats when administered orally in corn oil. Since halocarbons often occur as contaminants of drinking water, we sought to determine the influence of the vehicle, aqueous versus lipid, on the developmental toxicity of two of these agents. In separate assays, bromodichloromethane (BDCM) and carbon tetrachloride (CCl4) were administered by gavage to Fischer-344 rats on gestation days (GD) 6 15 at 0, 25, 50, or 75 mg/kg/day in either corn oil or an aqueous vehicle containing 10% Emulphor EL-620. Dams were allowed to deliver and the litters were examined postnatally. Uteri of females that did not deliver were stained with 10% ammonium sulfide to detect FLR. Effects of both agents on maternal weight gain were slightly more pronounced in the aqueous vehicle at lower doses, but at the highest dose, CCl4 was more maternally toxic in corn oil. Developmentally, both agents caused FLR at 50 and 75 mg/kg in both vehicles. At 75 mg/kg, dams receiving corn oil had significantly higher rates of FLR (83% for BDCM, 67% for CCl4) compared to their aqueous-vehicle counterparts (21% for BDCM, 8% for CCl4). Blood concentrations of BDCM following GD-6 gavage revealed a shorter elimination half-life in the aqueous dosing vehicle (2.7 h) compared to the oil vehicle (3.6 h). Benchmark doses of CCl4 were similar for the oil (18.9 mg/kg) and aqueous (14.0 mg/kg) vehicles. For BDCM, the corn oil vehicle yielded a less conservative (i.e., higher) value (39.3 mg/kg) than the aqueous vehicle (11.3 mg/kg), reflecting different confidence intervals around the estimated 5%-effect dose levels. PMID- 9398486 TI - Evaluation of effect profiles: Functional Observational Battery outcomes. AB - The Functional Observational Battery (FOB) is a neurotoxicity screening assay composed of 25-30 descriptive, scalar, binary, and continuous endpoints. These outcomes have been grouped into six biologically logical domains as a means to interpret the neuroactive properties of tested chemicals (V. C. Moser, 1992, J. Am. Coll. Toxicol. 10(6), 661-669). However, no data-based exploration of these functional domains has been done. We investigated the degree to which experimental data correspond to the domain groupings by examining severity scores from 10 chemicals tested using a standardized protocol for acute exposure (V. C. Moser et al., 1995, J. Toxicol. Environ. Health 45, 173-210) and identifying endpoint groupings (factors) that best describe the interrelationships in the data, allowing a statistical assessment of whether the FOB endpoints break into domains. We also used a standard measure of bivariate association to confirm the results of the factor analysis. Our results show that while there are clear relationships among variables that compose some domains, there is often substantial correlation among endpoints in different domains. In addition, we investigated a related issue concerning the relative power of the chosen endpoint groupings for identifying significant domain effects. Results from a randomization analysis of the 10 chemicals suggest that the neurophysiologic domain structuring may provide some degree of statistical efficiency for identifying effects. PMID- 9398487 TI - Cardiovascular effects after inhalation of large doses of albuterol dry powder in rats, monkeys, and dogs: a species comparison. AB - Albuterol is a quickly acting beta-2 adrenergic agonist bronchodilator widely used by asthmatics. Because recent case-control studies have suggested a relationship between the increase in mortality of asthmatics over the past decade and the use of beta 2-adrenergic agonists in the control of asthma, concern has developed regarding the potential cardiotoxicity of beta 2-specific adrenergic agonists, including albuterol. The aim of this investigation was to assess the potential for cardiotoxicity of inhaled albuterol dry powder in rats, monkeys, and dogs. All species were exposed to an aerosol of albuterol 1 h per day, 7 days per week, for at least 2 weeks. Control groups were exposed to filtered conditioned air and handled in the same manner as the albuterol-exposed animals. Plasma concentrations of albuterol confirmed systemic exposure. The daily inhaled dose received by the animals was calculated based on measured respiratory minute volumes, published respiratory tract deposition data, as well as HPLC-determined particle size distribution data and aerosolized albuterol concentrations. Multiples of the maximum daily clinical dose (presentation of 15 micrograms/kg in a 70-kg human) were approximately 0.25- to 2500-fold in the rat, 9- to 100-fold in the monkey, and 0.5- to 90-fold in the dog. No findings attributed to albuterol were observed in the monkey. Tachycardia and transient hypokalemia occurred in rats at multiples of 1.5 times or greater of the maximum clinical dose. Absolute and relative heart weights increased in rats receiving multiples of 47 times or greater of the maximum human dose. In the absence of histopathologic findings, the increases in rat heart weights were considered a physiologic hypertrophic response to tachycardia. In dogs tachycardia and transient hypokalemia occurred at all doses tested. Slight to mild fibrosis in the papillary muscles of the left ventricle of the heart occurred in dogs at multiples > or = 19 times the clinical dose. The cardiovascular effects observed were consistent with the known pharmacologic action of beta 2-adrenergic agonists. Due to the lack of toxicologically relevant findings in rats and monkeys and the wide safety margin in dogs, the findings in this study do not suggest a cardiotoxicity risk in the human population after repeated exposures to clinical doses of albuterol currently used in the treatment of asthma. PMID- 9398488 TI - Repeated independent exposures to domoic acid do not enhance symptomatic toxicity in outbred or seizure-sensitive inbred mice. AB - Domoic acid (DA) is an environmental neurotoxin to humans. This work examines whether repeated exposure to subsymptomatic or symptomatic nonlethal doses of domoic acid leads to enhanced symptomatic toxicity in ICR outbred and DBA inbred strains of laboratory mice. A multiple independent exposure paradigm was designed in which doses were administered intraperitoneally every other day for 7 days to achieve four separate exposures to domoic acid. We first examined the effect of repeated exposure on serum clearance of domoic acid. Serum domoic acid levels did not differ following a single or repeated exposure. We next examined the effect of repeated exposure on symptomatic toxicity. The mean toxicity scores did not show a significant difference between single and repeated exposures of either subsymptomatic (0.5 mg/kg) or symptomatic sublethal (2.0 mg/kg) doses of domoic acid. We then examined the effects of repeated domoic acid exposure on a second strain of mouse. DBA mice were chosen based upon their sensitivity to kainic acid induced seizures; however, the ICR mice were more sensitive to low-dose domoic acid toxicity, particularly in terms of onset and duration of stereotypic scratching behavior. Our results indicate that both strains of mice have comparable concentration-dependent toxic responses to domoic acid; however, differences exist in the magnitude of the response and in specific symptoms. The mean toxicity scores did not show a significant difference when a single exposure (1.0 and 2.0 mg/kg domoic acid) and repeated exposure of the same dose were compared in the DBA mice. This study provides no evidence that short-term repeated exposure to domoic acid in laboratory mice alters domoic acid clearance from the serum, or leads to a more sensitive or a greater neurotoxic response. PMID- 9398489 TI - Effects of acute and repeated exposures to Aroclor 1254 in adult rats: motor activity and flavor aversion conditioning. AB - While considerable research has focused on the neurotoxicity of developmental exposures to polychlorinated biphenyls, including Aroclor 1254, relatively little is known about exposures in adult animals. This study investigated the behavioral effects of acute and repeated Aroclor 1254 exposures to adult rats on motor activity and flavor aversion conditioning. Male Long-Evans rats (60 days old) were tested for motor activity in a photocell device after acute (0, 100, 300, or 1000 mg/kg, p.o.) or repeated (0, 1, 3, 10, 30 or 100 mg/kg/day, po, 5 days/week for 4 to 6 weeks exposure to Aroclor 1254. Motor activity was decreased dose dependently at doses of 300 mg/kg or more after acute exposure. Severe body weight loss and deaths occurred at 1000 mg/kg. Recovery of activity occurred over 9 weeks but was incomplete. After repeated exposure, motor activity was decreased dose-dependently at doses of 30 mg/kg or more, and severe weight loss and deaths occurred at 100 mg/kg. In contrast to acute exposure, complete recovery of activity occurred 3 weeks after exposure. Additional rats were water deprived (30 min/day) and received acute po administration of Aroclor 1254 (0, 10, 15, 25, 30, 100, or 300 mg/kg) shortly after consuming a saccharin solution. Three days later they were given the choice between consuming saccharin or water, and saccharin preferences were recorded. Saccharin preference was decreased at doses of 25 mg/kg or more. Additional experiments determined the effect of repeated saccharin Aroclor 1254 pairings (0, 3.75, 7.5, or 15 mg/kg/day, 14 days) followed by a choice test 1 day after the last dose. Repeated exposure to 15 mg/kg produced robust flavor aversion conditioning. Repeated exposure to 7.5 mg/kg produced flavor aversion conditioning in four of 12 rats. These results demonstrate that Aroclor 1254 causes hypoactivity and flavor aversions in adult rats; the no observable effect level (NOEL) for motor activity was 100 mg/kg for acute exposure and 10 mg/kg for repeated exposure for a period of up to 6 weeks. The acute NOEL for flavor aversion conditioning was 15 mg/kg while the repeated NOEL was 7.5 mg/kg. PMID- 9398490 TI - Trimethylphosphate: a 30-month chronic toxicity/carcinogenicity study in Wistar rats with administration in drinking water. AB - Trimethylphosphate (TMPO) was administered to 50 male and 50 female Wistar rats through their drinking water at doses of 0, 1, 10, or 100 mg/kg body weight up to 30 months. The dosage of 100 mg/kg was reduced to 50 mg/kg in week 54 for reasons of tolerance, and the animals were euthanized in week 100. Additional 10 animals per dose and sex were treated for 12 months and then euthanized for interim analysis. Weakness of the hind limbs, increased incidences of sunken flanks, distended abdomen, and poor general condition were observed in both sexes of the 100/50 mg/kg group beginning with week 46. Food intake was reduced in high dose males. At 10 mg/kg body weights were up to 10% (males) and at 100/50 mg/kg up to 20% (males) or 15% (females) lower than in controls. Mortality was not affected in animals receiving up to 10 mg/kg. At 100/50 mg/kg it was markedly increased, reaching about 70% at week 100. Relatively slight hematologic changes (reduced hemoglobin, hematocrit, erythrocyte counts, increased reticulocyte numbers, and thrombocyte counts as well as a shift in the differential blood count) at 100/50 mg/kg are interpreted as changes most probably secondary to the other toxic effects. Increased cholesterol concentrations in plasma, shifts in the serum protein electrophoresis (males), increased organ weights (females), and an increased incidence of necroses and lymphocytic infiltrations point to a treatment-related effect on the liver at 100/50 mg/kg. Slightly increased protein excretion, increased relative kidney weights, and an increased incidence of chronic progressive nephropathy are considered treatment-related but rather secondary effects at 100/50 mg/kg. At 100/50 mg/kg an increased incidence and severity of bilateral tubular atrophy in the testes was diagnosed. The most important toxic effect was neurotoxicity, consisting of degeneration and loss of nerve fibers in the peripheral nerves and the spinal cord, associated with myopathic changes, and occurring at 100/50 mg/kg. The no-observed-adverse-effect level, based on the suppression of body weight gain, is 1 mg/kg in males and 10 mg/kg in females. The incidence, time of occurrence, spectrum of types, and localizations of tumors provided no indication of a tumorigenic/carcinogenic effect of the test substance. TMPO is therefore considered not to be carcinogenic in Wistar rats. PMID- 9398491 TI - Two-generation reproductive toxicity study of dietary tributyl phosphate in CD rats. AB - Tributyl phosphate (TBP) was tested for reproductive toxicity in rats. Thirty weanlings/sex (F0) were exposed to TBP in the diet ad libitum at 0, 200, 700, or 3000 ppm for 10 weeks and then randomly mated within groups for 3 weeks with continued exposure. F0 parents and 10 F1 weanlings/sex/dose were necropsied, and adult reproductive organs, urinary bladders (both sexes), kidneys (males), and livers (females) were evaluated histologically. Thirty F1 weanlings/sex/dose continued exposure for 11 weeks and were bred as described above. F1 parents and F2 weanlings, 10/sex/dose, were then necropsied as described above. Adult toxicity was observed in both sexes and generations at 700 and 3000 ppm; observations included reduced body weights, weight gain and feed consumption, urinary bladder epithelial hyperplasia (both sexes), renal pelvis epithelial hyperplasia only at 3000 ppm (male kidneys), and centrilobular hypertrophy (female livers). At 200 ppm, transient reductions in body weight were observed in F0 and F1 females, with urinary bladder epithelial hyperplasia in F0 males and females and in F1 males. There was no evidence of reproductive toxicity, of reproductive organ pathology, or of effects on gestation or lactation at any dose tested. Postnatal toxicity was evidenced by consistent reductions in F1 and F2 pup body weights at 3000 ppm and by occasional weight reductions in F2 litters at 700 ppm, and was associated with maternal toxicity observed at these doses and times. Under the conditions of this study, a NOAEL was not determined for adult toxicity; the NOAEL for reproductive toxicity was at least 3000 ppm and the NOAEL for postnatal toxicity was approximately 200 ppm. PMID- 9398492 TI - Trihalomethane comparative toxicity: acute renal and hepatic toxicity of chloroform and bromodichloromethane following aqueous gavage. AB - Bromodichloromethane (BDCM) and chloroform (CHCl3) are by-products of drinking water chlorination and are the two most prevalent trihalomethanes (THMs) in finished drinking water. To date, no comprehensive comparison of the acute renal and hepatic effects of BDCM and CHCl3 following oral gavage in an aqueous dosing vehicle has been conducted. To characterize BDCM- and CHCl3-induced nephro- and hepatotoxicity following aqueous gavage and compare directly the responses between these THMs, 95-day-old male F-344 rats were given single oral doses of 0.0, 0.75, 1.0, 1.5, 2.0, or 3.0 mmol BDCM or CHCl3/kg body wt in an aqueous 10% Emulphor solution. Compound-related hepatic and renal damage was evaluated by quantitating clinical toxicity markers in the serum and urine, respectively. Both THMs appear to be equally hepatotoxic after 24 h, but BDCM caused significantly greater elevations in serum hepatotoxicity markers than CHCl3 at 48 h following exposure to 2.0 and 3.0 mmol/kg. In addition to causing more persistent liver toxicity than CHCl3, BDCM also appears to be slightly more toxic to the kidney at lower doses. Potency differences between the two THMs may be due to pharmacokinetic dissimilarities such as greater metabolism of BDCM to reactive metabolites or more extensive partitioning of BDCM into kidneys and fat depots, resulting in prolonged target tissue exposure. PMID- 9398493 TI - Single-dose and chronic dietary neurotoxicity screening studies on 2,4 dichlorophenoxyacetic acid in rats. AB - Forms of 2,4-dichlorophenoxyacetic acid (collectively known as 2,4-D) are herbicides used to control a wide variety of broadleaf and woody plants. Single dose acute and 1-year chronic neurotoxicity screening studies in male and female Fischer 344 rats (10/sex/dose) were conducted on 2,4-D according to the U.S. EPA 1991 guidelines. The studies emphasized a Functional Observational Battery (which included grip performance and hindlimb splay tests), automated motor activity testing, and comprehensive neurohistopathology of perfused tissues. Dosages were up to 250 mg/kg by gavage for the single-dose study, and up to 150 mg/kg/day in the diet for 52 weeks in the repeated-dose study. In the acute study, gavage with 250 mg/kg test material caused slight transient gait and coordination changes and clearly decreased motor activity at the time of maximal effect on the day of treatment (day 1). Mild locomotor effects occurred in one mid-dose rat (75 mg/kg), on Day 1 only. No gait, coordination, or motor activity effects were noted by day 8. In the chronic study, the only finding of neurotoxicologic significance was retinal degeneration in females in the high-dose group (150 mg/kg/day). Body weights of both sexes were slightly less than controls in the mid-dose group, and 10% less than controls in the high-dose group. In summary, the findings of these studies indicated a mild, transient locomotor effect from high-level acute exposure, and retinal degeneration in female rats from high level chronic exposure. Based on the results from these two studies, the no observed-adverse-effect level for acute neurotoxicity was 15 mg/kg/day and for chronic neurotoxicity was 75 mg/kg/day. PMID- 9398494 TI - Lack of embryotoxicity of fumonisin B1 in New Zealand white rabbits. AB - Fumonisin B1 (FB1) is one of a number of mycotoxins produced by fungi, especially Fusarium sp. As a contaminant of many maize-derived products, this toxin is associated with a variety of animal diseases, including esophageal cancer and possibly neural tube defects in humans. We have investigated the embryotoxic potential of this compound in New Zealand White rabbits. Animals were dosed by gavage daily on GD 3-19 with purified FB1 at 0.10, 0.50, or 1.00 mg/kg/day. Maternal lethality occurred at the 0.50 and 1.00 mg/kg/day doses. When examined on GD 29, there were no differences in maternal body weight, maternal weight gain, maternal organ weights, number of nonlive implantations, and number of malformations. Fetal weight was decreased at 0.50 and 1.00 mg/kg/day (13 and 16%, respectively); this was true for male and female pups. Fetal liver and kidney weights were also decreased at these doses. Analysis of embryonic sphinganine to sphingosine ratios demonstrated no differences between control and treated embryos on GD 20, although these ratios were increased in maternal urine, serum, and kidney when compared to control animals. These data suggest that FB1 did not cross the placenta and that the observed decreased fetal weight was probably the result of maternal toxicity, rather than any developmental toxicity produced by FB1. PMID- 9398495 TI - Expression of the immediate-early genes, c-fos, c-jun, and c-myc: a comparison in rats of nongenotoxic hepatocarcinogens with noncarcinogenic liver mitogens. AB - The involvement of the immediate-early (IE) genes c-fos, c-jun, and c-myc in regenerative liver hyperplasia is accepted, but their involvement in direct hyperplasia is uncertain. We have examined the hypothesis that the ability to induce IE genes may reflect the hepatocarcinogenic potential of a chemical. The ability of 1,4-dichlorobenzene (DCB) (300 mg/kg) (a noncarcinogenic rat liver mitogen), diethylhexyl phthalate (DEHP) (950 mg/kg), and chlorendic acid (120 mg/kg) (both nongenotoxic hepatocarcinogens) to induce c-fos, c-jun, and c-myc expression in rat liver was determined by Northern blot analysis and by in situ hybridization. Results were correlated to hepatic labeling index (LI) as determined by incorporation of BrdU in each of three lobes for each of three male F344 rats per group. Carbon tetrachloride (CCl4) (2 ml/kg) was used as a positive control. Increased LI was preceded by elevated expression of all three IE genes after CCl4, but also after DCB and DEHP, although induction by these was less marked. In all cases, there was considerable interanimal variation within groups, but little interlobe variation. Interestingly, there was a good correlation (r2 > or = 0.85) between c-myc expression and LI, but not between LI and c-fos or c jun. Despite the disparate carcinogenic potential of DEHP and DCB, both chemicals induced similar patterns of IE gene expression, suggesting that this cannot distinguish hepatocarcinogenic liver mitogens from noncarcinogenic liver mitogens. These data assist in the evaluation of IE gene expression both as a marker of direct versus regenerative hyperplasia and as an indicator of the hepatocarcinogenic potential of liver mitogens. PMID- 9398497 TI - Evaluation of the pre-, peri-, and postnatal toxicity of monoethanolamine in rats following repeated oral administration during organogenesis. AB - Pregnant Wistar rats (40/group) were administered monoethanolamine (MEA) as an aqueous solution by gavage at dose levels of 0, 40, 120, and 450 mg/kg/day on days 6 through 15 of gestation. On day 20 of gestation, 25 dams/group were euthanized and the fetuses were delivered by cesarean section, weighted, sexed, and examined for external, visceral, and skeletal alterations. The remaining dams (15/group) were allowed to litter and rear their pups to day 21 postpartum. The dams and pups were then euthanized and examined for gross pathologic changes. Gavage administration of 450 mg MEA/kg/day to pregnant rats resulted in maternal toxicity as evidenced by statistically significant (alpha = 0.05) decreases in feed consumption on gestation days 6-8 and 17-20 and on postpartum days 0-4. Additionally, statistically significant decreases in mean maternal body weights were observed on gestation days 15, 17, and 20 and on lactation days 0, 4, 7, and 21. Body weight gains of the 450 mg/kg/day dams were also significantly decreased (13% relative to controls) on gestation days 15-20. There was no evidence of maternal toxicity at 40 or 120 mg/kg/day of MEA. Despite the maternal effects observed at 450 mg/kg/day, no significant fetal effects were observed at this or any dose level tested, nor were there any indications of a treatment-related effect on postnatal growth or on the viability of offspring. Thus, it was concluded that MEA was not developmentally toxic to Wistar rats following repeated oral administration, even at maternally toxic dose levels as high as 450 mg/kg/day. PMID- 9398496 TI - The effects of perinatal/juvenile methoxychlor exposure on adult rat nervous, immune, and reproductive system function. AB - In order to address data gaps identified by the NAS report Pesticides in the Diets of Infants and Children, a study was performed using methoxychlor (MXC). Female rats were gavaged with MXC at 0, 5, 50, or 150 mg/kg/day for the week before and the week after birth, whereupon the pups were directly dosed with MXC from postnatal day (pnd) 7. Some dams were killed pnd7 and milk and plasma were assayed for MXC and metabolites. For one cohort of juveniles, treatment stopped at pnd21; a modified functional observational battery was used to assess neurobehavioral changes. Other cohorts of juveniles were dosed until pnd42 and evaluated for changes to the immune system and for reproductive toxicity. Dose dependent amounts of MXC and metabolites were present in milk and plasma of dams and pups. The high dose of MXC reduced litter size by approximately 17%. Ano genital distance was unchanged, although vaginal opening was accelerated in all treated groups, and male prepuce separation was delayed at the middle and high doses by 8 and 34 days, respectively. In the neurobehavioral evaluation, high dose males were more excitable, but other changes were inconsistent and insubstantial. A decrease in the antibody plaque-forming cell response was seen in males only. Adult estrous cyclicity was disrupted at 50 and 150 MXC, doses which also showed reduced rates of pregnancy and delivery. Uterine weights (corrected for pregnancy) were reduced in all treated pregnant females. High-dose males impregnated fewer untreated females; epididymal sperm count and testis weight were reduced at the high, or top two, doses, respectively. All groups of treated females showed uterine dysplasias and less mammary alveolar development; estrous levels of follicle stimulating hormone were lower in all treated groups, and estrus progesterone levels were lower at 50 and 150 MXC, attributed to fewer corpora lutea secondary to ovulation defects. These data collectively show that the primary adult effects of early exposure to MXC are reproductive, show that 5 mg/kg/day is not a NO(A)EL in rats with this exposure paradigm (based on changes in day of vaginal opening, pubertal ovary weights, adult uterine and seminal vesicle weights, and female hormone data) and imply that the sites of action are both central and peripheral. PMID- 9398498 TI - Co-response Coefficients, Monovalent Units, and Combinatorial Rules: Unification of Concepts in Metabolic Control Analysis AB - The Jacobian H of a linear metabolic pathway without feedback loops is tridiagonal. Its inverse, H-1, which is needed for calculating control coefficients or elasticities, can be decomposed into two regions of mutually dependent rows and columns. For each of these regions of H-1, all sub determinants of order two are zero. The existence of the two regions is shown to cause certain invariance properties of the ratios of control coefficients that can be expressed as co-response coefficients. Also the concept of monovalent functional units seems to be related to the existence of the regions. Moreover, the combinatorial rules for selecting the right modulations originate from the fact that certain sub-determinants of H-1 are zero when located within the regions. The regions of zero sub-determinants of order two of H-1 are thus a central element for the analysis of the linear pathway. The unifying potential of H-1 is not restricted to the linear chain but is also expected to be valid for complex metabolic systems.Copyright 1997 Academic Press Limited Copyright 1997 Academic Press Limited PMID- 9398499 TI - Self-organized huddles of rat pups modeled by simple rules of individual behavior. AB - Starting at infancy and continuing throughout adult life, huddling is a major component of the behavioral repertoire of Norway rats (Rattus norvegicus). Huddling behavior maintains the cohesion of litters throughout early life, and in adulthood, it remains a consistent feature of social behavior of R. norvegicus. During infancy, rats have severely limited sensorimotor capabilities, and yet they are capable of aggregating and display a form of group regulatory behavior that conserves metabolic effort and augments body temperature regulation. The functions of huddling are generally understood as group adaptations, which are beyond the capabilities of the individual infant rat. We show, however, that huddling as aggregative or cohesive behavior can emerge as a self-organizing process from autonomous individuals following simple sensorimotor rules. In our model, two sets of sensorimotor parameters characterize the topotaxic responses and the dynamics of contact in 7-day-old rats. The first set of parameters are conditional probabilities of activity and inactivity given prior activity or inactivity and the second set are preferences for objects in the infant rat's environment. We found that the behavior of the model and of actual rat pups compare very favorably, demonstrating that the aggregative feature of huddling can emerge from the local sensorimotor interactions of individuals, and that complex group regulatory behaviors in infant rats may also emerge from self organizing processes. We discuss the model and the underlying approach as a paradigm for investigating the dynamics of social interactions, group behavior, and developmental change. PMID- 9398500 TI - A discussion about the DiFrancesco-Noble model. AB - An obvious defect in the DiFrancesco-Noble (DN) model was found, so methods to overcome this inadequacy are put forward. Based on these methods, different kinds of modified DN model can be obtained, and numerical results show that most of the dynamics of the DN model can be essentially preserved by these modified systems. PMID- 9398501 TI - A conservation principle and its effect on the formulation of Na-Ca exchanger current in cardiac cells. AB - In this paper we show the presence of a latent conservation principle in the formulation of ionic currents in cardiac cells and examine its effect on a formulation of the sodium-calcium exchange current appearing in the Noble model of the sinoatrial node cell in the mammalian heart [see Noble et al. (1989) or Winslow et al. (1991)]. Our objective is to show that this formulation, if not corrected, will result in a serious instability in this cardiac cell model. In particular, under certain initial conditions, the solutions of the model equations will blow-up in finite time. We also propose a correction to the model equation for the sodium-calcium exchange current, and we show that the modified model agrees favorably with the original model. These phenomena also occur in the other cardiac cell models, such as those modeling the Purkinje fiber, the atrial cell and the ventricular cell. The changes proposed in this paper can be applied directly to these models as well. PMID- 9398502 TI - A Model for Cell Movement During Dictyostelium Mound Formation AB - Dictyostelium development is based on cell-cell communication by propagating cAMP signals and cell movement in response to these signals. In this paper we present a model describing wave propagation and cell movement during the early stages of Dictyostelium development, i.e. aggregation and mound formation. We model cells as distinct units whose cAMP relay system is described by the Martiel-Goldbeter model. To describe cell movement we single out three components: chemotactic motion, random motion and motion due to pressure between cells. This pressure result in cells crawling on top of each other and therefore to the extension of the aggregate into the third dimension. Using this model we are able to describe aggregation up to the mound stage. The cells in the mound move in a rotational fashion and their movement is directed by the counter-rotating spiral of the chemo-attractant cAMP. Furthermore, we show that the presence of two subpopulations with different inherent chemotactic velocities can lead to cell sorting in the mound. The fast moving cells collect into the centre while the slow cells occupy the rest of the mound. This model allows the direct comparison of the properties of the cAMP waves properties and movement behavior of individual cells with experimental data. Thereby it allows a critical test of our understanding of the basic cellular principles involved in the morphogenesis of a simple eukaryote.Copyright 1997 Academic Press Limited Copyright 1997 Academic Press Limited PMID- 9398503 TI - Sex, the Prisoner's Dilemma Game, and the evolutionary inevitability of cooperation. AB - It will be a universal feature of sexual populations that individuals prefer mates with typical rather than rare characteristics--essentially because most mutations reduce fitness. This is termed koinophilia. Koinophilia will also apply to behaviour. In particular, individuals will prefer mates that behave in social interactions that follow whatever rules are common in that population. Suppose that individuals interact in situations which can be represented by the Iterated Prisoner's Dilemma Game (IPD). If koinophilia is ignored, previous authors have shown that it is hard to find an evolutionarily stable strategy, and that strategies cycle indefinitely. However, if koinophilia is included, it has the effect of increasing the fitness of whatever happens to be the common strategy. This, in turn, has the effect of stabilizing almost any strategy (that has, for whatever reason, become the local norm) in the IPD. Different, partially isolated groups will thus become evolutionarily trapped in different behaviours, which are defended against alternative strategies originating through mutation or immigration. Groups that happen, by chance, to reach a cooperative strategy will be fitter, as groups, than those that reach defection (even though, in one-to-one encounters, it is the selfish individual who always wins). The ultimate result will be the replacement of selfish groups by cooperative groups. PMID- 9398504 TI - Pattern formation and spatiotemporal irregularity in a model for macrophage tumour interactions. AB - Solid tumours do not develop as a homogeneous mass of mutant cells, rather, they grow in tandem with normal tissue cells initially present, and may also recruit other cell types including lymphatic and endothelial cells. Many solid tumours contain a high proportion of macrophages, a type of white blood cell which can have a variety of effects upon the tumour, leading to a delicate balance between growth promotion and inhibition. In this paper we present a brief review of the main properties and interactions of such tumour-associated macrophages, leading to a description of a mathematical model for the spatial interactions of macrophages, tumour cells and normal tissue cells, focusing on the ability of macrophages to kill mutant cells. Analysis of the homogeneous steady states shows that, for this model, normal tissue is unstable to the introduction of mutant cells despite such an immune response, but that the composition of the resulting tumour can be significantly altered. Including random cell movement and chemical diffusion, we demonstrate the existence of travelling wave solutions connecting the normal tissue and tumour steady states, corresponding to a growing tumour, and of the development of a spatial instability behind the wave front. Numerical solutions are illustrated in one and two dimensions. We go on to estimate macrophage motility parameters using data from Boyden chamber experiments. We then extend our model to include macrophage chemotaxis, that is, their directed movement in response to gradients of chemicals secreted by tumours. Solutions in one dimension indicate the possibility of spatiotemporal irregularities within the growing tumour, which are deduced to be the result of a series of bifurcations as the effective domain length increases, leading to a permanently transient solution. These results suggest that tumour heterogeneity may arise, in part, as a natural consequence of the macrophage infiltration. Recent experiments suggest that macrophages may indeed be involved in spatiotemporal variations within some human tumours. PMID- 9398505 TI - Radiation damage accumulation over large time intervals: a descriptive model. AB - A simple model of DNA damage (mutation load) accumulation is considered. For time intervals exceeding average life duration, the kinetics of mutation load is determined. The major process determining these kinetics is shown to be natural selection (or any equivalent process of removal of the most affected lines). The characteristic time of this process (which is around three generations) is the approximate time of integration of the dose rate for determination of radiation damage. The conclusion is made that the maximum permissible dose should be established for this characteristic time, as it determines the remote effects of prolonged exposure to radiation. After this, the accumulated genetic damage presents "dose rate-effect" rather than "dose-effect" dependence. Dependence of mutation load on dose rate and parameters of selection processes is examined for different types of mutations. The cases of stable and exponentially decreasing dose rate are considered. The applicability of the model to human population is discussed. PMID- 9398506 TI - Single-beat estimation of the ventricular pumping mechanics in terms of the systolic elastance and resistance. AB - An elastance-resistance model has long been used to assess the systolic mechanical behavior of the ventricular pump under an in situ, open-chest experiment. However, there is difficulty in the clinical application of such a model because of the required isovolumetric signal that is obtained by occluding the ascending aorta in diastole. In this study, we determine the characteristics of an elastance-resistance model in the absence of isovolumeric measurement to quantify the physical properties of the left ventricle. A high-fidelity multisensor catheter was used to record the left ventricular pressure and ascending aortic flow in nine anesthetized, closed-chest dogs. The isovolumetric pressure was estimated from the instantaneous pressure of an ejecting contraction by a curve-fitting technique. Thus, the parameters in the characterization of systolic pumping mechanics could be inferred by the use of fitting this elastance resistance model. The results showed that the maximal systolic elastance was 7.3 +/- 2.8 mmHg and theoretical maximum flow was 494 +/- 194 ml s-1. These data were compatible with other reports in the literature. Moreover, in every dog studied the maximal systolic elastance was smaller than the end-systolic elastance which was determined by using the end-systolic pressure-volume relation. We suggested that an elastance-resistance model with the estimated isovolumetric pressure has the potential to measure the intrinsic systolic mechanics of the left ventricle in a closed-chest cardiovascular system. PMID- 9398507 TI - The adaptational system as a dynamical feedback system. AB - The characteristics of biological systems of adaptation are developed from the principle that the manifestations of life are modified by and must conform with their environment in order to enable organismic persistence. The two roles of the environment, termed "modifying" and "adaptive", give rise to the distinction of three elementary and sequentially coupled subsystems characterizing the adaptational system: the modifying system, comparator system, and state regulation system. The third system determines which state alterations are required for adaptation of manifestations (responses) to the demands of the adaptive environment. Adaptational valuation is introduced as a measure of adaptedness of response to adaptive environment. The dynamical aspect of adaptation is shown to be completely described by the timing of feedback. Discrete and continuous forms of adaptation can thus be treated on the same conceptional basis. All steps of the generic system formulation are illustrated with the help of a simple model with additive effects and linear state regulation. The system representation is used to demonstrate how basic intuitive conceptions, such as adaptational lag or adaptational capacity, can be made amenable to precise analysis. Another demonstration concerns recognition of adaptational clues resulting from the distinction between the two environmental classes, modifying and adaptive. Among these clues are challenges of low correlation between the two classes to adaptational systems and the specification of questions of the evolution of phenotypic plasticity. PMID- 9398508 TI - Modelling Bacterial Degradation of Organic Compounds with Genetic Networks AB - The bacterial degradation of organic compounds plays a crucial role in the biogeochemical cycles of the earth and in the clean-up of contaminated soils. The processes are carried out by bacterial consortia, rather than isolated strains, which are usually modelled by phenomenological kinetic equations which describe a fictitious, homogeneous bacterial species which mimics the behaviour of the consortium. An alternative modelling framework is presented here, where the bacterial consortia are considered as networks of genes interacting with other genes as well as with chemicals, which may be either introduced from outside or produced by bacterial metabolism. The model is based on an extension of the random Boolean network model of genetic networks, which makes use of continuous dynamical variables. Three different models are introduced, which differ in the way how they account for the existence of different species: (i) a single supercell model, where all the genes can interact strongly with each other; (ii) a graded interaction model, where genes interact strongly within a species, and weakly among different species; and (iii) a separate subsets model, where genes interact only within species. It is shown how this modelling framework is sound, as it is able to reproduce some of the generic behaviours of bacterial consortia, describing experimentally observed phenomena like population changes induced by contamination, and prey-predator dynamics.Copyright 1997 Academic Press Limited Copyright 1997 Academic Press Limited PMID- 9398509 TI - Activation of P2 late transcription by P2 Ogr protein requires a discrete contact site on the C terminus of the alpha subunit of Escherichia coli RNA polymerase. AB - Bacteriophage P2 late transcription requires the product of the P2 ogr gene. Ogr dependent transcription from P2 late promoters is blocked by certain point mutations affecting the alpha subunits of the host RNA polymerase. An alanine scan spanning the putative activation target in the alpha C-terminal domain (alphaCTD) was carried out to identify individual residues essential for Ogr dependent transcription from P2 late promoters. In addition, the effects of alanine substitutions in the regions of the alphaCTD previously reported to affect CAP-dependent activation of the lac promoter and UP-element DNA binding were examined. Residues E286, V287, L289 and L290 in helix 3, and residue L300 at the beginning of helix 4, define a surface-exposed patch on the alphaCTD important for Ogr-dependent activation. These residues, adjacent to the recently identified DNA-binding determinants, constitute a newly identified activation surface for protein:protein contact. Alanine substitutions at some of the residues that affect UP-element DNA binding also impaired activation. This suggests that upstream DNA-alpha contacts, in addition to alpha-Ogr contacts, may be important in P2 late transcription. Other residues implicated in the interaction of alpha with CAP are not required for activation by Ogr, consistent with previous genetic evidence suggesting that these activators contact different sites on the alphaCTD. PMID- 9398510 TI - Pactamycin resistance mutations in functional sites of 16 S rRNA. AB - Mutants of an archaeon Halobacterium halobium, resistant to the universal inhibitor of translation, pactamycin, were isolated. Pactamycin resistance correlated with the presence of mutations in the 16 S rRNA gene of H. halobium single rRNA operon. Three types of mutations were found in pactamycin resistant cells, A694G, C795U and C796U (Escherichia coli 16 S rRNA numeration) located distantly in rRNA primary structure but probably neighboring each other in the three-dimensional structure. Pactamycin resistance mutations either overlapped (C795U) or were located in the immediate vicinity of nucleotides protected by the drug in E. coli and H. halobium 16 S rRNA indicating that corresponding rRNA sites might be directly involved in pactamycin binding. Ribosomal functions were not affected significantly either by mutation of C795 (one of the positions protected by the P-site-bound tRNA), or by mutations of A694 and C796 (which neighbor nucleotides protected by tRNA) suggesting that tRNA-dependent protections of C795 and G693 are explained by a conformational change in the ribosome induced by the P-site-bound tRNA. A novel mode of pactamycin action is proposed suggesting that pactamycin restricts structural transitions in 16 S rRNA preventing the ribosome from adopting a functional conformation induced by tRNA binding. PMID- 9398511 TI - Crystal structure of annexin V with its ligand K-201 as a calcium channel activity inhibitor. AB - The crystal structure of recombinant human annexin V complexed with K-201, an inhibitor of the calcium ion channel activity of annexin V, was solved at 3.0 A by molecular replacement including the apo and high-calcium forms. K-201 was bound at the hinge region cavity formed by the N-terminal strand and domains II, III and IV, at the side opposite the calcium and membrane-binding surface, in an L-shaped conformation. Based on the complex and other annexin structures, K-201 is proposed to restrain the hinge movement of annexin V in an allosteric manner, resulting in the inhibition of calcium movement across the annexin V molecule. PMID- 9398512 TI - Structural characterization of the myoglobin active site using infrared crystallography. AB - We use polarized IR absorption on single crystals to determine the orientation of carbon monoxide bound at the active site of myoglobin, and conclude that the C-O bond lies approximately 7 degrees from the normal to the mean plane of the heme. This result disagrees with much larger angular displacements reported in structural models derived from X-ray and neutron diffraction measurements. The insensitivity of the IR-derived orientation to changes in pH or crystal packing contrasts with the wide variations in CO orientation among diffraction-based models and suggests that the latter are in error. The small energies required to displace the C-O bond 7 degrees from its energetically preferred upright geometry suggest that distortion of the surrounding protein, rather than the relatively undeformable Fe-C-O unit, is the main steric mechanism inhibiting CO binding to myoglobin. PMID- 9398513 TI - The central region of RepE initiator protein of mini-F plasmid plays a crucial role in dimerization required for negative replication control. AB - The RepE protein (251 residues, 29 kDa) of mini-F plasmid, mostly found as dimers, plays a key role in mini-F replication. Whereas monomers bind to the origin to initiate replication, dimers bind to the repE operator to repress its own transcription. Among the host factors required for mini-F replication, a set of molecular chaperones (DnaK, DnaJ and GrpE) is thought to facilitate monomerization of RepE dimers. To further understand the structural basis of functional differentiation between the two forms of RepE, we examined the region(s) critical for dimerization by isolation and characterization of RepE mutants that were defective in autogenous repressor function. Such mutations were isolated from two separate regions of RepE, the central region (residues 111 to 161) and the C-terminal region (residues 195 to 208). The central region overlapped the region where the chaperone-independent copy-up mutations were previously isolated (residues 93 to 135). Likewise the mini-F mutant plasmids, carrying the mutations in the central region, could replicate in a dnaK null mutant host. One of them, S111P (111th serine changed to proline), showed a very high origin-binding activity vis-a-vis a severely reduced operator-binding activity, much like the RepE54 (R118P) mutant previously shown to form only monomers. Gel filtration and chemical crosslinking studies with purified RepE revealed that S111P primarily formed monomers, whereas other mutant proteins formed mostly dimers. On the other hand, analysis of deletion mutants revealed that the N-terminal 42 and the C-terminal 57 residues were dispensable for dimerization. Thus, the region spanning residues 93 to 161 of RepE (including Ser111 and Arg118) appeared to be primarily involved in dimerization, contributing to the negative regulation of plasmid replication. PMID- 9398514 TI - Structure-function correlations in the XerD site-specific recombinase revealed by pentapeptide scanning mutagenesis. AB - Xer-mediated site-specific recombination contributes to the stability of circular chromosomes in bacteria by resolving plasmid multimers and chromosome dimers to monomers prior to cell division. Two related site-specific recombinases, XerC and XerD, each catalyse one pair of strand exchange during Xer recombination. In order to relate the recently determined structure of XerD to its function, the XerD protein was subjected to pentapeptide scanning mutagenesis, which leads to a variable five amino acid cassette being introduced randomly into the target protein. This has allowed identification of regions of XerD involved in specific DNA binding, in communicating with the partner recombinase, XerC, and in catalysis and its control. The C-terminal domain of XerD, comprising two-thirds of the protein, contains the catalytic active site and comprises ten alpha helices (alphaE to alphaN) and a beta hairpin. A flexible linker connects this domain to the N-terminal domain that comprises four alpha helices (alphaA to alphaD). Pentapeptide insertions into alphaB, alphaD, alphaG, or alphaJ interfered with DNA binding. Helices alphaG and alphaJ comprise a pseudo helix turn-helix DNA binding motif that may provide specificity of recombinase binding. An insertion in alphaL, adjacent to an active site arginine residue, led to loss of cooperative interactions between XerC and XerD and abolished recombination activity. Other insertions close to active site residues also abolished recombination activity. Proteins with an insertion in the beta hairpin turn bound DNA, interacted cooperatively with XerC and had a phenotype that is consistent with the protein being defective in XerD catalysis. This beta hairpin appears to be highly conserved in related proteins. Insertions at a number of dispersed locations did not impair XerD catalytic activity or DNA binding, but failed to allow XerC catalysis in vivo, indicating that several sites of interaction between XerD and XerC may be important for activation of XerC catalysis by XerD. PMID- 9398515 TI - Rules governing the orientation of the 2'-hydroxyl group in RNA. AB - Molecular dynamics simulations reveal that, in C3'-endo sugar puckers, only three orientations are accessible to the 2'-hydroxyl groups distinctive of RNA molecules: towards (i) the O3', (ii) the O4' of the same sugar, and (iii) the shallow groove base atoms. In the rarer C2'-endo sugar puckers, orientations towards the O3' atom of the same sugar are strongly favoured. Surprisingly, in helical regions, the frequently suggested intra-strand O2'-H(n)...O4'(n+1) interaction is not found. This observation led to the detection of an axial C H...O interaction between the C2'-H2'(n) group and the O4'(n+1) atom contributing to the stabilization of RNA helical regions. Subsequent analysis of crystallographic structures of both RNA and A-DNA helices fully supports this finding. Specific hydration patterns are also thought to play a significant role in the stabilization of RNA structures. In the shallow groove of RNA, known as a favourable RNA or protein-binding region, three well-defined hydration sites are located around the O2' atom. These hydration sites, occupied by water molecules exchanging with the bulk, constitute, after dehydration, anchor points for specific interactions between RNA and nucleic acids, proteins or drugs. Therefore, the fact that the 2'-hydroxyl group is not monopolised by axial stabilization, together with its water-like behaviour, facilitates complex formation involving RNA helical regions. PMID- 9398516 TI - X-ray fibre diffraction study of an elevated temperature structure of poly(dA).poly(dT). AB - A reversible conformational transition between two discrete double helical forms of poly(dA).poly(dT) has been put into evidence by X-ray fibre diffraction. We observed that the transition between the well known B' conformation and a new helical structure (B*) occurs at a relative humidity near 80%, when the temperature is raised above 30 degrees C. It appears that the B* conformation is not just a distorted B' form of poly(dA).poly(dT) but rather a stable (up to a least 70 degrees C) distinct double helical structure of that polynucleotide. Analysis of X-ray patterns allowed us to present the geometrical parameters of a molecular model of this new double helix. It consists of 11.4 nucleotide pairs per turn in a pitch length of about 36.7 A. The proposed high-temperature right handed helical structure of poly(dA).poly(dT) is a member of the B-DNA family since the duplex has C1'-exo furanoses in both antiparallel but geometrically identical sugar-phosphate strands. The present finding may shed light on interpretations of results obtained from premelting or nucleosome formation processes involving (dA.dT) tracts in synthetic or natural DNA polymers. PMID- 9398517 TI - Structure and dynamics of the iron responsive element RNA: implications for binding of the RNA by iron regulatory binding proteins. AB - The iron responsive element (IRE) is a approximately 30 nucleotide RNA hairpin that is located in the 5' untranslated region of all ferritin mRNAs and in the 3' untranslated region of all transferrin receptor mRNAs. The IREs are bound by two related IRE-binding proteins (IRPs) which help control intracellular levels of iron by regulating the expression of both ferritin and transferrin receptor genes. Multi-dimensional NMR and computational approaches were used to study the structure and dynamics of the IRE RNA in solution. The NMR data are consistent with formation of A-form helical stem regions, a one-base internal bulge and a Watson-Crick C.G base-pair between the first and fifth nucleotides in the loop. A superposition of refined structures indicates that the conserved C in the internal bulge, and three residues in the six-nucleotide hairpin loop are quite dynamic in this RNA. The structural roles of the stems, the loop and the bulge in the function of the IRE RNA and in possible interactions with the iron regulatory protein are discussed. PMID- 9398518 TI - The structures of human glutathione transferase P1-1 in complex with glutathione and various inhibitors at high resolution. AB - The human pi-class glutathione S-transferase (hGST P1-1) is a target for structure-based inhibitor design with the aim of developing drugs that could be used as adjuvants in chemotherapeutic treatment. Here we present seven crystal structures of the enzyme in complex with substrate (glutathione) and two inhibitors (S-hexyl glutathione and gamma-glutamyl- (S-benzyl)cysteinyl-D phenylglycine). The binding of the modified glutathione inhibitor, gamma-glutamyl (S-benzyl)cysteinyl-D-phenylglycine, has been characterized with the phenyl group stacking against the benzyl moiety of the inhibitor and making interactions with the active-site residues Phe8 and Trp38. The structure provides an explanation as to why this compound inhibits the pi-class GST much better than the other GST classes. The structure of the enzyme in complex with glutathione has been determined to high resolution (1.9 to 2.2 A) in three different crystal forms and at two different temperatures (100 and 288 K). In one crystal form, the direct hydrogen-bonding interaction between the hydroxyl group of Tyr7, a residue involved in catalysis, and the thiol group of the substrate, glutathione, is broken and replaced by a water molecule that mediates the interaction. The hydrogen-bonding partner of the hydroxyl group of Tyr108, another residue implicated in the catalysis, is space-group dependent. A high-resolution (2.0 A) structure of the enzyme in complex with S-hexyl glutathione in a new crystal form is presented. The enzyme-inhibitor complexes show that the binding of ligand into the electrophilic binding site does not lead to any conformational changes of the protein. PMID- 9398519 TI - The RNA binding domain of ribosomal protein L11: three-dimensional structure of the RNA-bound form of the protein and its interaction with 23 S rRNA. AB - The three-dimensional solution structure has been determined by NMR spectroscopy of the 75 residue C-terminal domain of ribosomal protein L11 (L11-C76) in its RNA bound state. L11-C76 recognizes and binds tightly to a highly conserved 58 nucleotide domain of 23 S ribosomal RNA, whose secondary structure consists of three helical stems and a central junction loop. The NMR data reveal that the conserved structural core of the protein, which consists of a bundle of three alpha-helices and a two-stranded parallel beta-sheet four residues in length, is nearly the same as the solution structure determined for the non-liganded form of the protein. There are however, substantial chemical shift perturbations which accompany RNA binding, the largest of which map onto an extended loop which bridges the C-terminal end of alpha-helix 1 and the first strand of parallel beta sheet. Substantial shift perturbations are also observed in the N-terminal end of alpha-helix 1, the intervening loop that bridges helices 2 and 3, and alpha-helix 3. The four contact regions identified by the shift perturbation data also displayed protein-RNA NOEs, as identified by isotope-filtered three-dimensional NOE spectroscopy. The shift perturbation and NOE data not only implicate helix 3 as playing an important role in RNA binding, but also indicate that regions flanking helix 3 are involved as well. Loop 1 is of particular interest as it was found to be flexible and disordered for L11-C76 free in solution, but not in the RNA-bound form of the protein, where it appears rigid and adopts a specific conformation as a result of its direct contact to RNA. PMID- 9398520 TI - Ligand-induced conformational changes in ras p21: a normal mode and energy minimization analysis. AB - A normal mode and energy minimization of ras p21 is used to determine the flexibility of the protein and the origin of the conformational differences between GTP and GDP-bound forms. To preserve the integrity of the structures, a hydration shell of water molecules was included as part of the system. Certain low-frequency modes were found to have high involvement coefficients with the conformational transition between the GTP and GDP-bound structures; the involvement coefficients of some of the modes increase when the gamma-phosphate group is removed. Two unstable modes that appear in the GTP-bound structure upon deletion of the gamma-phosphate group were determined and shown to have dominant contributions in the regions of switch I and switch II; there was also a significant displacement of loop 1. The initial motion in these regions is predicted by the modes to be approximately perpendicular to the direction of the transition from the GTP-bound state to the GDP-bound state. The overall conformational change in the switch I and II regions involves rearrangements of the protein backbone within these regions, rather than rigid body motion. Differences in the low-frequency modes of the GTP and GDP-bound forms appear to play a role in ligand binding. A coupling between the helix alpha3 position and the deletion of the gamma-phosphate group may be involved in the interaction with GAP. The oncogenic mutation G12D leads to a global increase in the rigidity of the protein. Thus, the mutant is likely to have a higher barrier for the conformational change to the inactive form; this would slow the transition and could be related to its oncogenic properties. PMID- 9398521 TI - Contribution of water molecules in the interior of a protein to the conformational stability. AB - Water molecules frequently occur in the interior of globular proteins. To elucidate the contribution of buried water molecules to the conformational stability of a protein, we examined the crystal structures and the thermodynamic parameters of denaturation of six Ile to Ala/Gly mutant human lysozymes, in which a cavity is created at each mutation site by the substitution of a smaller side chain for a larger one. One or two ordered water molecules were found in the cavities created in some mutants (I106A, I59A and I59G). The cavity volumes for these three mutants were bigger than those that remained empty in the other mutants. The stability of the mutant proteins with the newly introduced water molecules was about 8 kJ/mol higher than that expected from the change in hydrophobic surface area (DeltaDeltaASAHP) exposed upon denaturation. It was concluded that a water molecule in a cavity created in the interior of a protein contributes favorably to the stability. PMID- 9398522 TI - Clarification of the dimerization domain and its functional significance for the Escherichia coli nucleoid protein H-NS. AB - The Escherichia coli nucleoid protein, H-NS, functions as a global regulator for expression of a wide variety of genes. We recently analyzed the structure function relationship of H-NS with special reference to the domains responsible for transcriptional repression and DNA-binding, respectively. However, identification of the presumed dimerization domain of H-NS and its functional significance was elusive. To address this particular issue, we first examined a set of N-terminally or C-terminally truncated forms of H-NS, in terms of their so called dominant-negative effect on the in vivo function of the wild-type H-NS. The results showed that certain truncated forms exhibit such a dominant-negative effect, but others did not. As judged by the results of the dominant-negative effect, it was assumed that a relatively central portion of H-NS extending from residues 21 to 63 is involved in dimerization. This was confirmed by an in vitro chemical cross-linking analysis and a gel filtration analysis with these truncated forms of H-NS. Furthermore, the use of the dominant-negative phenotype, caused by a truncated form of H-NS (named N91), allowed us to isolate a missense mutant, which was expected to be specifically defective in dimerization. This mutant had an amino acid substitution at position 30 (Leu30 to Pro) in N91 consisting of the N-terminal 91 amino acids of H-NS. This mutant was indeed defective in the in vitro ability to form a heterodimer with the wild-type H-NS. When this particular single amino acid substitution was introduced into the full length H-NS, the resultant H-NS mutant had lost the ability to form dimers in vitro and to function as a transcriptional repressor. These findings collectively provided us with evidence that the ability of H-NS to form a dimer is crucial for H-NS to function as a transcriptional repressor. PMID- 9398523 TI - NMR analysis of main-chain conformational preferences in an unfolded fibronectin binding protein. AB - A 130-residue fragment of the Staphylococcus aureus fibronectin-binding protein has been found to exist in a highly unfolded conformation at neutral pH. Measurement of experimental NMR 3JHNalpha coupling constants provides evidence for individual residues having distinct main-chain conformational preferences that are dependent both on the amino acid concerned and on neighbouring residues in the sequence. Analysis shows that these variations in the populations of individual residues can be explained in detail in terms of statistical distributions of conformational states derived from the protein data base. In particular, when the preceding residue has a beta-branched or aromatic side chain, a significant increase occurs in the population of the less sterically restricted b region of phi,psi space. The results indicate that the local structure of the fibronectin binding protein in solution, under conditions where it displays full activity, approximates very closely to a statistical random coil structure. This may be an important feature in the biological role of this and other polypeptides involved in protein-protein interactions. PMID- 9398524 TI - Assembly of the N-dependent antitermination complex of phage lambda: NusA and RNA bind independently to different unfolded domains of the N protein. AB - The N protein of bacteriophage lambda activates expression of the delayed early genes of this phage by modifying RNA polymerase (RNAP) into a form that is resistant to termination signals. N binds to the boxB hairpin that forms in the nascent RNA transcript upon transcription of the nut regulatory element, and then interacts with RNAP by RNA looping. The binding of the N-boxB subassembly to the transcription complex is further stabilized by interaction with the Escherichia coli NusA protein. N, free in solution, exists as an unfolded protein that becomes partially structured upon binding specifically to boxB RNA. Because NusA does not assist in antitermination unless N is specifically bound to boxB, we have asked whether the structural change induced by binding to boxB affects the interaction of N with NusA. Using fluorescence spectroscopy, we have measured the affinity of N for NusA in the presence and absence of boxB RNA. We find that NusA binds to the unfolded N protein with a dissociation constant (Kd) of approximately 70 nM, and although N undergoes a significant structural change upon binding to boxB, the binding affinity of NusA for a N protein complexed with boxB is not altered. We have also shown that the boxA element of nut does not affect NusA binding to N-boxB. These results demonstrate that the interaction of N with NusA is independent of RNA binding, arguing that NusA must interact with an unfolded region of the polypeptide that remains unstructured even when N binds to boxB RNA. To further establish this point we isolated a truncated peptide containing the amino-terminal 36 residues of the N protein. Binding of boxB RNA to this peptide showed that all of the structural change in N that occurs upon binding to boxB RNA is localized within the amino-terminal 36 residues of N, therefore the C terminus of N, including the regions necessary for NusA binding and RNAP activation, remains unfolded when the full length N binds to boxB RNA. Thus it appears that N can be described as an unfolded multi-domain protein that becomes ordered in a modular fashion as it encounters its various binding partners within the N-dependent antitermination complex. PMID- 9398525 TI - SelD homolog from Drosophila lacking selenide-dependent monoselenophosphate synthetase activity. AB - The isolation and molecular characterization of an invertebrate gene that encodes a homolog of the human selenophosphate synthetase 1 is described. This Drosophila gene, termed selD-like, is located in the cytogenetic interval 50 D/E on the right arm of chromosome 2. It is expressed ubiquitously throughout embryogenesis and found to be highly enriched in the developing gut and in the nervous system of the embryo. The SelD-like from Drosophila was purified after expression in Escherichia coli. The purified protein does not catalyze the selenide-dependent ATP hydrolysis reaction and its gene does not complement a selD lesion in E. coli. These results and the fact that selD-like possesses an arginine residue at the position of the essential Cys17 (E. coli nomenclature) indicate that the Drosophila gene exerts a function different from that of the classical selenophosphate synthetases. Two classes of SelD proteins can therefore be differentiated. The class I proteins contain a cysteine or selenocysteine residue in the active site and display selenide-dependent selenophosphate synthetase activity. Class II proteins, including Drosophila selD-like and human selenophosphate synthetase 1 are devoid of this activity and they possess other amino acids in position 17. PMID- 9398526 TI - Stress-induced duplex DNA destabilization in scaffold/matrix attachment regions. AB - S/MARs are DNA elements 300 to several thousand base-pairs long, which are operationally defined by their affinity for the nuclear scaffold or matrix. S/MARs occur exclusively in eukaryotic genomes, where they mediate several functions. Because S/MARs do not have a clearcut consensus sequence, the characteristics that define their activity are thought to be structural. Ubiquitous S/MAR binding proteins have been identified, but to date no unique binding sequence or structural motif has been found. Here we show by computational analysis that S/MARs conform to a specific design whose essential attribute is the presence of stress-induced base-unpairing regions (BURs). Stress induced destabilization (SIDD) profiles are calculated using a previously developed statistical mechanical procedure in which the superhelical deformation is partitioned between strand separation, twisting within denatured regions, and residual superhelicity. The results of these calculations show that BURs exhibit a succession of evenly spaced destabilized sites that would render part or all of the S/MAR sequence single stranded at sufficient superhelicity. These analyses are performed for a range of sequenced S/MAR elements from the borders of eukaryotic gene domains, from centromeres, and from positions where S/MARs are known to support the action of an enhancer. The results reported here are in excellent agreement with earlier in vitro chemical reactivity studies. This approach demonstrates the potential for computational analysis to predict the points of division of the eukaryotic genome into functional units (domains), and also to locate certain cis-regulatory sequences. PMID- 9398527 TI - Inter-domain cross-linking and molecular modelling of the hairpin ribozyme. AB - The hairpin ribozyme is a small catalytic RNA composed of two helical domains containing a small and a large internal loop and, thus, constitutes a valuable paradigm for the study of RNA structure and catalysis. We have carried out molecular modelling of the hairpin ribozyme to learn how the two domains (A and B) might fold and approach each other. To help distinguish alternative inter domain orientations, we have chemically synthesized hairpin ribozymes containing 2'-2' disulphide linkages of known spacing (12 or 16 A) between defined ribose residues in the internal loop regions of each domain. The abilities of cross linked ribozymes to carry out RNA cleavage under single turnover conditions were compared to the corresponding disulphide-reduced, untethered ribozymes. Ribozymes were classed in three categories according to whether their cleavage rates were marginally, moderately, or strongly affected by cross-linking. This rank order of activity guided the docking of the two domains in the molecular modelling process. The proposed three-dimensional model of the hairpin ribozyme incorporates three different crystallographically determined structural motifs: in domain A, the 5'-GAR-3'-motif of the hammerhead ribozyme, in domain B, the J4/5 motif of group I ribozymes, and connecting the two domains, a "ribose zipper", another group I ribozyme feature, formed between the hydroxyl groups of residues A10, G11 of domain A and C25, A24 of domain B. This latter feature might be key to the selection and precise orientation of the inter-domain docking necessary for the specific phosphodiester cleavage. The model provides an important basis for further studies of hairpin ribozyme structure and function. PMID- 9398528 TI - An interaction between a specified surface of the C-terminal domain of RecA protein and double-stranded DNA for homologous pairing. AB - RecA protein and its homologs catalyze homologous pairing of dsDNA and ssDNA, a critical reaction in homologous genetic recombination in various organisms from a virus, microbes to higher eukaryotes. In this reaction, RecA protein forms a nucleoprotein filament on ssDNA, which in turn binds to naked dsDNA for homology search. We suggested that the C-terminal domain of RecA protein plays a role in capturing the dsDNA. Here, we isolated the C-terminal domain as a soluble form and determined the solution structure by NMR spectroscopy. The overall folding of the NMR structure agrees with that of the corresponding part of the reported crystal structure, but a remarkable difference was found in a solvent-exposed region due to intermolecular contacts in the crystal. Then, we studied the interaction between the C-terminal domain and DNA, and found that significant chemical shift changes were induced in a specific region by titration with dsDNA. SsDNA induced a much smaller chemical shift perturbation. The difference of DNA concentrations to give the half-saturation of the chemical shift change showed a higher affinity of the C-terminal region toward dsDNA. Combined with our previous results, these provide direct evidence that the defined region in the C-terminal domain furnishes a binding surface for DNA. PMID- 9398529 TI - Three-dimensional structure of diferric bovine lactoferrin at 2.8 A resolution. AB - The three-dimensional structure of diferric bovine lactoferrin (bLf) has been determined by X-ray crystallography in order to investigate the factors that influence iron binding and release by transferrins. The structure was solved by molecular replacement, using the coordinates of diferric human lactoferrin (hLf) as a search model, and was refined with data to 2.8 A resolution by simulated annealing (X-PLOR) and restrained least squares (TNT). The final model comprises 5310 protein atoms (residues 5 to 689), 124 carbohydrate atoms (from ten monosaccharide units, in three glycan chains), 2 Fe3+, 2 CO32- and 50 water molecules. This model gives an R-factor of 0.232 for 21440 reflections in the resolution range 30.0 to 2.8 A. The folding of the bLf molecule is essentially the same as that of hLf, but bLf differs in the extent of closure of the two domains of each lobe, and in the relative orientations of the two lobes. Differences in domain closure are attributed to amino acid changes in the interface, and differences in lobe orientations to slightly altered packing of two hydrophobic patches between the lobes. Changed interdomain interactions may explain the lesser iron affinity of bLf, compared with hLf, and two lysine residues behind the N-lobe iron site of bLf offer new insights into the "dilysine trigger" mechanism proposed for iron release by transferrins. The bLf structure is also notable for several well-defined oligosaccharide units which demonstrate the structural factors that stabilise carbohydrate structure. One glycan chain, attached to Asn545, appears to contribute to interdomain interactions and may modulate iron release from the C-lobe. PMID- 9398530 TI - The native and the heat-induced denatured states of alpha-chymotrypsinogen A: thermodynamic and spectroscopic studies. AB - We report the first protein phase-diagram characterized by a combination of volumetric, calorimetric, and spectroscopic techniques. More specifically, we use ultrasonic velocimetry, densimetry, and differential scanning calorimetry, in conjunction with UV absorbance and CD spectroscopy to detect and to characterize the conformational transitions of alpha-chymotrypsinogen A as a function of both pH and temperature. As judged by the CD spectra, we find that, at room temperature, the protein remains in the native state over the entire pH range investigated (pH 1 to 10). The melting profiles of the native state reveal three distinct pH domains in which protein denaturation produces different final states. Below pH 3.1, we find the heat-induced denatured state of the protein to be molten globule (MG), lacking the native-like tertiary structure, while exhibiting significant secondary structural elements. At neutral and alkaline pH, we find the heat-induced denatured state to be unfolded (U), lacking both tertiary and secondary structures, while being structurally similar to the urea unfolded state. At intermediate pH values (between pH 3.1 and 7), we find the heat-induced denatured state to exhibit properties characteristic of both the MG and U states. Although at room temperature the protein remains native within the whole pH range studied (pH 1 to 10), our volumetric data reveal that the native state slightly "softens" at low pH, probably, due to pH-induced alterations in electrostatic forces causing the packing of the protein interior at low pH and room temperature to become less "tight". This softening of the protein at low pH is reflected in an 8% increase in the intrinsic compressibility, kM, of the protein "native" state. Our volumetric data also allow us to conclude that the heat-induced MG state retains a liquid-like, water-inaccessible core, with a volume that corresponds to about 40% of the solvent-inaccessible core of the native state. By contrast, our volumetric data are consistent with the U state of the protein being essentially unfolded, with the majority of its constituent atomic groups being solvent exposed and, therefore, strongly hydrated. PMID- 9398532 TI - Folding of barnase in the presence of the molecular chaperone SecB. AB - SecB is a molecular chaperone dedicated to interact exclusively with proteins destined for translocation across membranes. We find that SecB interacts with barnase during its folding in a similar manner to its interaction with GroEL. On mixing acid-denatured barnase with SecB in a stopped-flow spectrofluorimeter under conditions that favour refolding, we observe a series of fluorescence changes, corresponding to the binding of the denatured protein and the subsequent refolding of multiply and singly bound forms. The different phases were assigned using a combination of kinetics and mutant proteins. The refolding of barnase when bound to SecB is strongly retarded but never blocked. Multiply bound barnase is less tightly bound and refolds with a higher rate constant than singly bound barnase. Up to 4 mol of denatured barnase bind to 1 mol of tetrameric SecB. PMID- 9398531 TI - Electrostatic and non-electrostatic contributions to the binding free energies of anthracycline antibiotics to DNA. AB - The knowledge about molecular factors driving simple ligand-DNA interactions is still limited. The aim of the present study was to investigate the electrostatic and non-electrostatic contributions to the binding free energies of anthracycline compounds with DNA. Theoretical calculations based on continuum methods (Poisson Boltzmann and solvent accessible surface area) were performed to estimate the binding free energies of five selected anthracycline ligands (daunomycin, adriamycin, 9-deoxyadriamycin, hydroxyrubicin, and adriamycinone) to DNA. The free energy calculations also took into account the conformational change that DNA undergoes upon ligand binding. This conformational change appeared to be very important for estimating absolute free energies of binding. Our studies revealed that the absolute values of all computed contributions to the binding free energy were quite large compared to the total free energy of binding. However, the sum of these large positive and negative values produced a small negative value of the free energy around -10 kcal/mol. This value is in good agreement with experimental data. Experimental values for relative binding free energies were also reproduced for charged ligands by our calculations. Together, it was found that the driving force for ligand-DNA complex formation is the non-polar interaction between the ligand and DNA even if the ligand is positively charged. PMID- 9398533 TI - C-capping and helix stability: the Pro C-capping motif. AB - Here we have performed a statistical analysis of the protein database to find new putative local C-terminal motifs in alpha-helices. Our analysis shows that certain combinations of X-Pro pairs (Asn, Cys, His, Phe, Tyr, Trp, Ile, Val and Leu), in which residue X is the C-cap and the Pro is at position C', are more abundant than expected. In those pairs, except for the aliphatic residues, the presence of the Pro residue at C' tends to restrict the phi and psi dihedral angles of the residue at position C-cap, around -130 degrees , 70 degrees , respectively. For the aromatic residues as well as for His, the chi1 angle is around -60 degrees and the edge of the His and aromatic rings are close to the carbonyl group of the residue i - 4. In all the pairs having the above dihedral angles for residue C-cap, the main-chain amino group of Pro at C' is close to the last three main-chain carbonyls of the alpha-helix. The above structural arrangements suggests the existence of a stabilising electrostatic interaction of the residues at positions C-cap and C' with the helix macrodipole. We have denominated this putative local motif, the Pro-capping motif. To asses its importance in helix stability we have analysed by nuclear magnetic resonance (NMR) and far-UV circular dichroism (CD) a set of polyalanine-based peptides containing two of the above pairs: His-Pro and Phe-Pro, as well as the corresponding controls. In the case of the His-Pro pair we have found NMR evidence for the formation of the Pro-capping motif in aqueous solution. CD analysis shows that the presence of a Pro residue alters the C-cap properties of the preceding amino acids in the case of His and Phe makes them more favourable. The Pro-capping motif with the appropriate sequence, determines the location of the C terminus of alpha-helices and stabilises the helical conformation having Pro as the C' residue. PMID- 9398534 TI - Hydroxylamine treatment increases glutathione-protein and protein-protein binding in human erythrocytes. AB - Hydroxylamine is a direct-acting hematotoxic agent leading to hemolytic anemia in animals and man. The effect of hydroxylamine on the morphology, sulfhydryl status and membrane skeletal proteins of human erythrocytes were studied. Loss of reduced glutathione (GSH) from the red blood cells was directly proportional to the hydroxylamine concentration used. This loss of GSH was larger than the sum of the increase in the amounts of extracellular glutathione and intracellular oxidized glutathione (GSSG). The extracellular glutathione is mainly present as GSSG, which is in agreement with the fact that only GSSG is exported from the erythrocytes by membrane bound ATPases. Lack of GSSG export was not limited by decreased ATP levels in the erythrocytes and we concluded that the GSH that disappeared did not become available as intracellular GSSG. After reduction of the erythrocyte incubates the lost GSH was almost completely recovered indicating that the lost GSH is present in the cell as protein-glutathione mixed disulfides. Glutathione thus stored within the cell can be quickly recovered by combined thioltransferase and glutathione reductase activity when conditions become more favorable again. SDS-polyacrylamide gel electrophoresis of membrane ghosts from human red cells revealed changes in skeletal proteins with a smearing of bands 1, 2 and 3 to the higher molecular weight end of the gel and the appearance of new monomeric and dimeric hemoglobin bands at about 16 and 30 kD. The observed alterations are probably a consequence of disulfide bridge formation between cellular proteins (mainly hemoglobin) and skeletal proteins as well as between hemoglobin monomers. Exposure of hydroxylamine to erythrocytes caused severe Heinz body formation but the outside morphology of the cells was only marginally altered. The described changes in sulfhydryl status of the red blood cells are likely to play a major role in the premature splenic sequestration of hydroxylamine-damaged erythrocytes. PMID- 9398535 TI - A novel interferon-inducible gene expressed during myeloid differentiation. AB - The acute promyelocytic leukemia cell line, NB4, can be induced to differentiate to mature granulocytes by retinoic acid treatment. A novel retinoic acid inducible cDNA clone, designated RI58, was isolated from a cDNA library constructed from retinoic acid-treated NB4 cells by differential hybridization. RI58 cDNA encodes a protein of 58kDa which has a similarity in its amino acids sequence to interferon (IFN)-inducible proteins. In addition, RI58 was induced by recombinant human IFN-alpha (rhIFN-alpha) in NB4 cells. RI58 was detectable within 4 hours post-stimulation with rhIFN-alpha, while it took as long as 1day after retinoic acid stimulation. Culture supernatant from retinoic acid-treated NB4 cells also induced RI58 expression similarly as rhIFN-alpha. This activity in culture supernatant was inhibited by anti-leukocyte IFN antiserum which showed specific reactivity to rhIFN-alpha. These results indicate that RI58 is induced by retinoic acid stimulation through autocrinally secreted IFN-alpha from NB4 cells. In the retinoic acid-treated NB4 cells, the expression of RI58 was increased along the process of differentiation. On the other hand, it was expressed constitutively in untreated non-hematopoietic cell lines and mature hematopoietic cell lines. PMID- 9398536 TI - Structural and functional analysis of the Pig-a protein that is mutated in paroxysmal nocturnal hemoglobinuria. AB - There is now convincing evidence that the Pig-a gene is mutated in patients with paroxysmal nocturnal hemoglobinuria (PNH), a disease in which one or more clones of hematopoietic cells have incomplete assembly of glycosylphosphatidylinositol (GPI) anchors and absence of GPI-linked protein expression on the cell surface. Little is known, however, about the Pig-a protein product that is necessary for GPI anchor bioassembly. Relatively few substitution (missense) Pig-a gene mutations have been identified, but we noted two apparent clusters at codons 128 129 and 151-156 and hypothesized that these might represent critical regions of the Pig-a protein. We therefore used site-directed mutagenesis to create conservative mutations in the Pig-a protein, then performed structural and functional analysis. Each Pig-a mutation generated a Pig-a protein of normal size and stability, but certain mutations had substantial deleterious effects on protein function. Conservative mutation of codons histidine 128 (H128), serine 129 (S129), and serine 155 (S155) had greatly diminished function, while mutations of flanking residues had no effect on function. Our results represent the first structure/function analysis of the Pig-a protein, and suggest that codons H128, S129, and S155 represent critical regions of the Pig-a protein. Our results also suggest a means by which transgenic mice with a "partial knock-out" of Pig-a function could be generated, which would allow investigation of PNH in an animal model. PMID- 9398538 TI - An exonic polymorphism in the human glucose phosphate isomerase (GPI) gene. AB - A polymorphic site has been found in the human glucose phosphate isomerase (GPI) gene. This site is produced by a A-->G substitution at nt 489 of GPI cDNA, resulting in a silent mutation. To our knowledge, it is the first defined polymorphism in the gene. It is present with similar gene frequencies in Asian, American White, African American, and Jewish populations. PMID- 9398537 TI - Expression of alpha IIb beta 3 integrin (GPIIb-IIIa) in myeloid cell lines and normal CD34+/CD33+ bone marrow cells. AB - Regulation of myeloid cell proliferation and differentiation in the bone marrow is mediated, in part, by the interaction of integrins on early myeloid cells with the extracellular matrix proteins secreted by stromal cells. To further define adhesive protein receptors of early myeloid cells, we examined the expression of the integrin GPIIb-IIIa (alphaIIbbeta3) in leukemic cell lines KG-1a, KG-1, and HL-60, that represent early stages of myeloid differentiation. All three cell lines expressed surface GPIIb-IIIa as measured by flow cytometry and by binding of 125I-anti-GPIIb-IIIa monoclonal antibody. Preincubation of cells with human AB serum or platelet releasate increased GPIIb-IIIa surface expression. GPIIb transcripts were identified in all three cell lines by Northern blot analysis. Furthermore, we readily detected GPIIb transcripts in fluorescence activated cell sorted (FACS) myeloid cells from normal human bone marrow by RT-PCR. Cloning and sequencing of the PCR products established the identity of GPIIb transcripts in the leukemic cell lines and CD34+/CD33+ normal bone marrow cells. Since the normal myeloid cells also demonstrated markers corresponding to the maturational stage of KG-1a/KG-1 cells, we propose that GPIIb-IIIa may serve as a myeloid differentiation antigen and as a key integrin of myeloid precursors. PMID- 9398539 TI - Air-Broadening and Shift Coefficients of O3 Lines in the nu2 Band and Their Temperature Dependence AB - Room temperature measurements of self- and air-broadening coefficients are reported for over 370 transitions covering a range of 0 BH + BH out of various electronic states of B2H2 is studied. The nature of binding in B2H2 is discussed in terms of the composition of the electronic wavefunctions. Copyright 1997Academic Press PMID- 9398545 TI - Laser Spectroscopy of Vibration-Rotation Lines in the 3 <-- 0, 5 <-- 0, and 6 <-- 0 Overtones of HBr AB - The Doppler-limited vibration-rotation absorption spectrum of HBr in the near infrared wavelength region was measured using a GaAs semiconductor laser and a ring-type titanium sapphire laser. About 100 lines in the v = 3 <-- 0, 5 <-- 0, and 6 <-- 0 bands were assigned and 16 Dunham coefficients Y10-Y50, Y01-Y41, Y02 Y22, Y03-Y13, and Y04 for H79Br and H81Br species were determined by a global least-squares fit using the v = 0, v = 1 --> 0, and v = 2 --> 1 lines by V. Braun and P. F. Bernath (J. Mol. Spectrosc. 167, 282-287, 1994), the v = 7 <-- 0 lines by P. Bernage and P. Niay (C. R. Acad. Sci. Paris Ser. B 282, 243-245, 1976), and the v = 3 <-- 0, v = 5 <-- 0, and v = 6 <-- 0 lines by this work. Copyright 1997Academic Press PMID- 9398546 TI - High Resolution Study of the 3nu2, nu1 + nu2, and nu2 + nu3 Bands of H2Te AB - High-resolution Fourier transform spectra of a natural sample of hydrogen telluride and of monoisotopic H2130Te have been recorded in the 3.2-4-0 MUm spectral region where the 3nu2, nu1 + nu2, and nu2 + nu3 bands of this molecule absorb. The (030) rotational levels were least-squares fitted using a Watson-type Hamiltonian whereas it proved necessary to consider the strong Coriolis interaction coupling the (110) and the (011) rotational levels. In this way all the experimental levels were calculated to within their experimental uncertainty and precise sets of vibrational energies and rotational and coupling constants were obtained for the (030), (110), and (011) vibrational states of H2130Te, H2128Te, H2126Te, H2125Te, H2124Te, H2123Te, and H2122Te. The band centers for the most abundant isotopic species, namely H2130Te, are:nuo(3nu2) = 2565.4428, nuo(nu1 + nu2) = 2911.4098, nuo(nu2 + nu3) = 2915.9599 cm-1 Copyright 1997Academic Press PMID- 9398547 TI - Analysis of the 2nu1 + nu2 + 2nu3 Band of Ozone AB - The 2nu1 + nu2 + 2nu3 band of ozone, occurring in the 4780 cm-1 region, has been observed for the first time, using a Fourier transform spectrometer, at 0.008 cm 1 resolution and using a large path length pressure product. Assignments of vibration-rotational transitions have been made up to J = 48 and Ka = 9. As a few levels with Ka = 1 or 2 are perturbed, it has been necessary to take into account the Coriolis resonance between the (212) and (141) vibrational states. With the effective Hamiltonian explicitly accounting for the interaction between these two states, the fit on 165 energy levels leads to the rms deviation of 1.9 x 10(-3) cm-1, which is near the experimental accuracy. Line intensities of the 2nu1 + nu2 + 2nu3 band have been measured and calculated. The set of spectroscopic parameters for interacting bands, as well as transition moment constants, is given. A complete list of line positions and intensities, with a cutoff of 1 x 10(-26) cm-1/molecule.cm-2 at 296 K up to J = 65 and Ka = 15, has been generated, which leads to the integrated band intensity Sv (2nu1 + nu2 + 2nu3) = (5.1 +/- 2.0) x 10(-23) cm-1/molecule.cm-2. Copyright 1997Academic Press PMID- 9398548 TI - The High-Resolution FTIR Spectrum of Jet-Cooled CH3CF2Cl AB - The infrared spectrum of HCFC-142b, CH3CF2Cl, has been recorded at high and low resolution using a supersonic jet-FTIR spectrometer system. Molecular parameters for a number of vibrational states have been obtained from rovibrational analyses of the C-type bands nu14 (1192 cm-1) and nu15 (967 cm-1), the A-type band nu7 (905 cm-1), and the B-type bands nu5 (1234 cm-1) and nu7 + nu11 (1215 cm-1). A rotational temperature of ca. 50 K in the jet expansion has been estimated from a comparison of the experimental spectrum with band simulations calculated using the resultant molecular constants. Copyright 1997Academic Press PMID- 9398549 TI - Direct Measurements of Line-Mixing Coefficients in the nu1 + nu2 Q Branch of CO2 AB - High-resolution measurements of the (nu1 + nu2) Q branch of pure CO2 were made using a difference frequency spectrometer with resolution of 5 x 10(-5) cm-1 and a signal-to-noise ratio of 2000:1. Lines Q(2) through Q(32) were measured with up to 14 lines in a single spectrum. The analysis of the branch has been performed on data taken at 301 K and pressures less than 11 kPa. The spectra were analyzed on a line-by-line basis within the Rosenkranz approximation of weak overlap [P. W. Rosenkranz, IEEE Trans. Antennas Propagation AP-23, 498 (1975)]. The lineshape profile included Doppler broadening and Dicke narrowing [R. H. Dicke, Phys. Rev. 89, 472 (1953)] using a modified hard collision model [S. G. Rautian and I. I. Sobel'man, Sov. Phys. Uspekh. 9, 701 (1967)] with line mixing. For each line the broadening, Dicke narrowing, and line-mixing coefficients were determined. The broadening coefficients are in good agreement with measurements of lines belonging to different CO2 bands. Our measured line-mixing parameters are compared to those predicted by a relaxation matrix which was calculated from an exponential power gap (EPG) law [L. L. Strow, D. C. Tobin, and S. E. Hannon, J. Quant. Spectrosc. Radiat. Transfer 52, 281 (1994)]. The vibrational band intensity and the linear pressure shift of the branch were also measured. Copyright 1997Academic Press PMID- 9398550 TI - Ab initio Calculated Rotation-Vibration Linestrengths for HeH2+ AB - Together with the recently determined potential energy surface for the ground electronic state of HeH2+ [V. Spirko and W. P. Kraemer, J. Mol. Spectrosc. 172, 265-274 (1995)], the electric dipole moment components were calculated directly as expectation values with the corresponding length operators at the center of mass of the ion and using the variationally optimized configuration interaction wavefunctions. From the fitted potential energy and dipole moment functions all bound rotation-vibration energy levels and the line strengths of all dipole allowed bound-bound transitions were evaluated variationally within the framework of the Sutcliffe-Tennyson Hamiltonian. Strong transitions, especially for the (He...H2)+ stretching motion, were obtained in the 500-800 cm-1 infrared frequency range. The present calculations demonstrate that the conditions for detecting the still unobserved rotation-vibration spectrum of HeH2+ are rather promising. Copyright 1997Academic Press PMID- 9398551 TI - High-Resolution Infrared Spectrum of Vinyl Fluoride at 500 cm-1 AB - The infrared spectrum of vinyl fluoride (CH2=CHF) has been investigated in the region 390-590 cm-1 at room temperature and at a resolution of 0.0016 cm-1 using a Fourier transform spectrometer. The rovibrational analysis of the spectral features allowed us to assign about 11 000 lines (J X2A" Transition of HO2: Analysis of the 000 000 Band AB - The near-infrared emission spectrum of the A2A' --> X2A" transition of the hydroperoxyl radical, HO2, has been studied by Fourier transform spectrometry. The 000 --> 000 band has been recorded at high spectral resolution. DeltaKa = +/ 1 subbands up to Ka' = 9 --> Ka" = 8 and Ka' = 8 --> Ka" = 9 comprising lines from rotational levels up to N' = 32 have been observed. With about a factor of 10 lower intensity, DeltaKa = 0 subbands 0-0 to 6-6 were found which are shown to be due to magnetic dipole transitions. In addition, in the 2-2 sub-band "forbidden" electric dipole lines were observed. These likely are induced by Renner-Teller interaction. Local perturbations extending over 3-10 N' values are found in the Ka' = 0-7 levels of the A2A' state. One series of perturbations is attributed to DeltaKa = 0, DeltaJ = 0 interactions with the 112 level of the X2A" ground state. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9398569 TI - Investigation of C-O Stretching Band of CD3OH: Assignments of Far-Infrared Laser Lines AB - We have investigated the high-resolution Fourier transform spectrum of the C-O stretching fundamental band of CD3OH in order to assign far-infrared (FIR) laser transitions. The absorption spectrum was analyzed by means of the "Ritz" program, which calculates the energy level values directly from the Rydberg-Ritz combination principle. We have also used the "LaseRitz" program to facilitate the assignment of the FIR laser lines. As a consequence we could determine 12 new assignments, confirming 4 previously proposed ones and predicting new FIR laser emissions. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9398570 TI - Near-Infrared Diode Laser Spectroscopy of the Nitrogen Molecule in Rydberg State: Analysis of the c1Piu - a"1Sigmag+, v = 1-0 Band AB - A new singlet-singlet absorption band between Rydberg states of the nitrogen molecule was studied by near-infrared diode laser spectroscopy in the 1.3 &mgr;m region. An analysis was made for the band to establish line assignments and determine molecular parameters for both the lower and the upper vibronic states. As a result of the analysis, this band was assigned to the c1Piu-a"1Sigmag+, v = 1-0 band. The anomaly of the Lambda-type doubling structure in the c1Piu (v = 1) state was discussed in connection with the interaction with the b'1Sigmau+ (v = 4) state. The predissociation in the c1Piu (v = 1) state was also discussed relating with the line broadening observed. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9398571 TI - The 3nu1 + nu2 Combination Band of HOCl: Assignments, Perturbations, and Line Intensities AB - The high-resolution spectra (0.012 cm-1) of the 3nu1 + nu2 combination band of hypochlorous acid HO35(37)Cl in the near infrared ( approximately 11 478 cm-1) have been measured using a titanium:sapphire intracavity laser absorption (ICLA) spectrometer. Line assignments, absolute intensities, and the total band intensity for both isotopomers are reported. In the course of the band analysis two Ka branches (Ka = 2,3) were found to be perturbed via low-order Fermi-type (anharmonic) resonances by a dark perturber which has been identified to be the 2nu1 + 2nu2 + 3nu3 state. The data are compared with intensity predictions from simple empirical models and discussed with regard to detection limits for this molecule in the near infrared spectral region of the atmosphere. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9398572 TI - Lambda-Splittings and Line Intensities in the First Overtone of Nitric Oxide AB - Using Fourier transform spectra, absolute line parameters (positions, Lambda splittings, and intensities) have been measured in the two allowed subbands 2Pi1/2-2Pi1/2 and 2Pi3/2-2Pi3/2 of the 2 <-- 0 vibrational band of the 14N16O molecule. These line parameters have been compared with previous experimental results and with the values available in the current HITRAN database. The measured line intensities led to the determination of the transition dipole moment as well as the Herman-Wallis coefficients. Improved spectroscopic constants were also obtained for the v = 2 vibrational state. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9398573 TI - Intensity Measurements of Deltal > 1 Transitions of Several Isotopomers of HCN AB - The intensities of the forbidden Q-branch transitions 02(2f)0-00(0)0, 12(2f)0 00(0)0, and 02(2f)1-00(0)0 for HCN have been measured. The intensities of the 02(2f)0-00(0)0 transitions of DCN, D13C15N, and H12C15N were also measured, as well as the 02(2f)1-00(0)0 transitions of H12C15N and H13C15N. These Q-branch transitions are forbidden even when the effects of l-type resonance are considered so they must get their intensity from some other Coriolis interactions. The much stronger P- and R-branch lines for the e levels of these same vibrational transitions were also measured and they are shown to get most of their intensity from l-type resonance. However, the same Coriolis resonance that gives intensity to the Q-branch transitions seems to affect the DeltaJ = ±1 transitions as shown by the difference in the intensities of the DeltaJ = +1 and DeltaJ = -1 transitions. Measurements of the intensity of the 03(3e)0 00(0)0 and 03(3f)0-00(0)0 transitions shows that they derive most, but perhaps not all, of their intensity from l-type resonance. An unsuccessful search for forbidden DeltaJ = 0, e-e transitions for the strong 10(0)0-00(0)0 band shows that there is no detectable mixing of the e and f levels. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9398574 TI - Nitrogen Broadening of Absorption Lines in the nu1, nu2, and nu3 Bands of H2S Determined by Applying an IR Diode Laser Spectrometer AB - With an infrared diode laser spectrometer in pulse mode we studied the N2 broadening of H2S absorption lines in the nu1, nu2, and nu3 bands. Altogether 22 lines were investigated: 16 lines from the R branch of the nu1 band (3 X1 summation operator+ System PMID- 9398583 TI - LETTER TO THE EDITOR PMID- 9398582 TI - LETTER TO THE EDITOR PMID- 9398584 TI - Reliability, distribution, and validity of age-related cognitive deficits in the Morris water maze. AB - In the present study, F-344 rats throughout 1.5 to 26 months of age were tested in the reference memory version, a moving-platform repeated acquisition version, and in a cued platform version of the Morris water maze. The results suggest that: (1) performance in the water maze declines continuously, beginning at the earliest age, and very closely fits a linear function; (2) there are robust, reliable differences between individuals in terms of their performance in the Morris water maze, but chronological age accounts for only a fraction of the variance between individuals; (3) there is no evidence of a bimodal distribution among aged rats--there is no distinct subgroup of individuals that performs so poorly that they are qualitatively different from the majority of the population, and distinctions between "impaired" and "unimpaired" subjects must be based on arbitrary criteria that may not be consistent from one study to the next; (4) age related deficits in the Morris water maze may not be restricted to learning and memory, but may also include deficits in attention, the ability to process spatial information, and/or the ability to develop efficient spatial search strategies; and (5) swim distance is the most appropriate measure of cognitive function in the Morris water maze, but the relationship between this measure and other measures of noncognitive function make it clear that swim distance may not be a pure measure of cognitive function. Although the Morris water maze remains a valuable preclinical test with better validity and specificity than many other behavioral tests, measures of performance in the Morris water maze should not be considered synonymous with cognitive function. PMID- 9398585 TI - Epinephrine effects on memory are not dependent on hepatic glucose release. AB - Epinephrine released or administered soon after a given training task modulates memory processes. Since epinephrine does not readily cross the blood-brain barrier, studies have suggested that some of the central effects of epinephrine might be mediated by peripheral release of glucose. These experiments examined the involvement of blood glucose levels in the posttraining effects of peripherally administered epinephrine. The effects of the administration of epinephrine (25 and 625 microg/kg) [corrected] on memory of an inhibitory avoidance task were evaluated in fed and fasted rats (depleted glycogen stores in liver). Blood glucose levels after the task in each group were also measured. Female Wistar rats were divided in two groups. Fed and 48-h-fasted animals were submitted to the inhibitory avoidance task and received i.p. epinephrine or saline immediately after training. The test session was carried out 48 h after training. Epinephrine (25 or 625 microg/kg) [corrected] caused an increased glycemia in fed rats, but no effect was observed in fasted animals. Administration of epinephrine 25 microg/kg [corrected] induced a facilitation of memory, while epinephrine 625 microg/kg [corrected] impaired retention (either in fasted or in fed animals). There was no relation between increased glycemia induced by epinephrine and its effects on memory, since this drug presented its classical effects independently of the previous state of the animal (fed or fasted). The results of the present study suggest that the effects of systemic released or administered epinephrine on memory processes are not dependent on hepatic glucose release. PMID- 9398586 TI - Effects of dark rearing and light exposure on memory for a passive avoidance task in day-old chicks. AB - Light exposure during embryogenesis is necessary for functional and morphological maturation in the domestic chick. In the present study, dark incubation was demonstrated to induce a weak amnestic effect on retention for a passive avoidance task and a diminution in discriminative memory ability in day-old chicks. Putative explanations based on possible motor, attentional, or visual impairment were excluded. Light exposure of dark-reared eggs, specifically during embryonic days E19 to E20, alleviated the retention and discrimination deficits. The processes which might mediate between prehatch light stimulation and posthatch behavioral effects are discussed. PMID- 9398587 TI - Rotation of water in the Morris water maze interferes with path integration mechanisms of place navigation. AB - The idea that place navigation in the Morris water maze is implemented by path integration between locations determined by landmark sighting was investigated in a 200-cm-diameter pool in which circular (7.2 degrees/s) motion of water could be induced by tangentially arranged water jets. The rats were trained at 8 trials per day to navigate to an erectable platform which was raised after the rat had spent a criterion time in the target annulus (30 cm in diameter) in the midpoint of the NW quadrant. Asymptotic escape latency of 7 s was reached after 9 days in moving water (n = 8) and after 6 days in stationary water (n = 8). The group overtrained for 13 days in stable water performed well even after it was transferred to moving water. Changing the sense of rotation of water from counterclockwise to clockwise did not affect the asymptotic performance. The above findings show that overtrained rats rely on landmark sighting rather than on path integration. The influence of water movement reappeared when place navigation to a new target (SW) was examined in alternating 2-s periods of light (L) and darkness (D). On the first day, the latencies were 15.2 +/- 1.2 and 22.8 +/- 1.9 s in stable and moving water, respectively, but dropped to 10 s on the following day. The tracks generated in the L period were more tortuous than those generated in the D period and this difference was more pronounced in moving than in stable water. It is concluded that path integration mechanisms supporting navigation during intervals of darkness are impaired in moving water but that this impairment disappears in overtrained animals. PMID- 9398588 TI - Intracerebroventricular infusion of the NMDA receptor-associated glycine site antagonist 7-chlorokynurenate impairs water maze performance but fails to block hippocampal long-term potentiation in vivo. AB - Most previous studies investigating the relationship between N-methyl-D-aspartate receptor-dependent synaptic plasticity and learning have employed drugs that either compete with glutamate for access to the primary agonist binding site (e.g., D-2-amino-5-phosphopentanoic acid) or block the associated ion channel (e.g., dizocilpine). This study targeted the glycine receptor site located on the NMDA receptor complex. Chronic intracerebroventricular infusion of the glycine site antagonist 7-chlorokynurenate (7CK; 75 mM, 0.5 microliter/h, icv, for up to 14 days) impaired performance of male Lister hooded rats during acquisition of a spatial reference memory task in the water maze. In addition, however, these animals showed sensorimotor deficits, including a prolonged righting reflex, ataxia, and difficulty in staying on the escape platform. On completion of behavioral testing, the rats were anesthetized with urethane and an attempt was made to induce LTP in the hippocampus ipsilateral to the infusion cannula. Both control and 7CK-infused animals displayed equivalent long-term potentiation (LTP) 60 min posttetanus. A novel analytical technique for assaying drug tissue levels involving high-performance liquid chromotography with fluorescence detection revealed that tissue levels of 7CK in hippocampus were extremely low and unlikely to be sufficient to affect LTP, as observed. These findings neither support nor compromise the LTP/learning hypothesis, but they illustrate some of the problems of using drugs to elucidate the neurobiological mechanisms of learning and memory and the importance of a within-subjects design incorporating behavioral, physiological, and biochemical measures. PMID- 9398589 TI - Synelfin regulation during the critical period for song learning in normal and isolated juvenile zebra finches. AB - Male zebra finches (Taeniopygia guttata) learn to sing during a critical period in adolescence. We previously described a presynaptic protein, synelfin, whose mRNA is increased early in this critical period in a brain nucleus specifically implicated in song learning, lateral MAN (lMAN). In the current study, in situ hybridization was used to map this change in gene expression to the subregion of lMAN that projects to the robust nucleus of the archistriatum (RA), the principal motor output of the telencephalic circuit that controls song production. Using confocal immunofluorescence microscopy, we detected numerous puncta of synelfin immunoreactivity that apparently represent presynaptic terminals from lMAN in the RA of young males. Synelfin immunoreactivity in RA declined abruptly between 40 and 45 days of age, a time of major synaptic reorganization in RA. This change did not occur until about 10 days after the decline in synelfin mRNA in cell bodies within lMAN, indicating a relatively slow turnover of the protein in presynaptic terminals and suggesting that some of the functional changes that occur during the critical period may arise from regulatory decisions that were initiated a week or more earlier. Depriving birds of tutoring did not halt or delay the decline of synelfin mRNA in lMAN. This change in gene expression must not be a consequence of early song learning, but may reflect an innate or programmed step in song circuit development. PMID- 9398590 TI - Memory formation: the sequence of biochemical events in the hippocampus and its connection to activity in other brain structures. AB - Recent data have demonstrated a biochemical sequence of events in the rat hippocampus that is necessary for memory formation of inhibitory avoidance behavior. The sequence initially involves the activation of three different types of glutamate receptors followed by changes in second messengers and biochemical cascades led by enhanced activity of protein kinases A, C, and G and calcium calmodulin protein kinase II, followed by changes in glutamate receptor subunits and binding properties and increased expression of constitutive and inducible transcription factors. The biochemical events are regulated early after training by hormonal and neurohumoral mechanisms related to alertness, anxiety, and stress, and 3-6 h after training by pathways related to mood and affect. The early modulation is mediated locally by GABAergic, cholinergic, and noradrenergic synapses and by putative retrograde synaptic messengers, and extrinsically by the amygdala and possibly the medial septum, which handle emotional components of memories and are direct or indirect sites of action for several hormones and neurotransmitters. The late modulation relies on dopamine D1, beta-noradrenergic, and 5HT1A receptors in the hippocampus and dopaminergic, noradrenergic, and serotoninergic pathways. Evidence indicates that hippocampal activity mediated by glutamate AMPA receptors must persist during at least 3 h after training in order for memories to be consolidated. Probably, this activity is transmitted to other areas, including the source of the dopaminergic, noradrenergic, and serotoninergic pathways, and the entorhinal and posterior parietal cortex. The entorhinal and posterior parietal cortex participate in memory consolidation minutes after the hippocampal chain of events starts, in both cases through glutamate NMDA receptor-mediated processes, and their intervention is necessary in order to complete memory consolidation. The hippocampus, amygdala, entorhinal cortex, and parietal cortex are involved in retrieval in the first few days after training; at 30 days from training only the entorhinal and parietal cortex are involved, and at 60 days only the parietal cortex is necessary for retrieval. Based on observations on other forms of hippocampal plasticity and on memory formation in the chick brain, it is suggested that the hippocampal chain of events that underlies memory formation is linked to long-term storage elsewhere through activity-dependent changes in cell connectivity. PMID- 9398591 TI - 2-Deoxy-D-galactose effects on passive avoidance memorization in the rat. AB - 2-Deoxy-D-galactose (do-gal) hinders glycoprotein fucosylation. This compound was intracerebroventricularly administered to male adult Wistar rats in order to assess whether it could exert amnesic effects on a passive avoidance response (PAR) to be learned in the light-dark box apparatus. Three experiments were performed. In the first, do-gal was administered immediately after the acquisition trials at three dosages (2, 4, and 8 mumol). It was found that only the 4-mumol dosage was followed by PAR disruption. In the second, do-gal was administered at the adequate dosage (4 mumol) either 30 min before the acquisition trial or 30 min before retrieval testing. It was found that only the preretrieval administration was followed by PAR impairment. In the third, do-gal (4 mumol) was administered in postacquisition, at increasing postacquisition delays (0.25, 1.5, 4, and 6 h). It was found that there was PAR disruption only after do-gal administration at the shortest (0.25 h) delay. The results confirm that in the rat, glycoprotein fucosylation is involved in some of the phases of memory trace processing, and they are discussed in relation to other findings in the rat and the chick. PMID- 9398592 TI - A brain of her own: a neural correlate of song assessment in a female songbird. AB - The song control region in the avian forebrain is a series of discrete, interconnected nuclei mediating song learning and production. It has been studied in males or in species where both sexes sing. Little is known about the neural correlates of song perception in nonsinging females, often the intended recipients of song. We studied cowbirds (Molothrus ater), a species in which only males sing but in which females discriminate between males on the basis of song. We focused on nucleus lMAN because it has been implicated in early song acquisition, a stage relevant to both sexes to choose among competing acoustic models. We found that volume of lMAN was monomorphic in cowbirds. Moreover, the volume and neuronal number of female lMAN were positively correlated with selectivity of copulatory responding. The results provide strong evidence of nonsinging female's use of "song" control nuclei for song perception without the possibility of song production. PMID- 9398593 TI - Arecoline via miniosmotic pump improves AF64A-impaired radial maze performance in rats: a possible model of Alzheimer's disease. AB - Male Sprague-Dawley rats, preoperatively trained in a 1-h delay non-match-to position radial maze task, received bilateral stereotaxic injections of a selective cholinotoxin, ethylcholine aziridinium ion (AF64A: 3 nmol/3 microliters/lateral ventricle). Animals treated with AF64A made significantly more total postdelay errors than vehicle controls. Sustained delivery, via miniosmotic pumps, of arecoline (0.1, 0.3, 1, 3, 10, or 30 mg/kg/day sc for 14 days) attenuated the AF64A-induced cognitive impairment in a dose-dependent manner, producing an inverted U-shaped dose-response function which was optimal at 1.0 mg/kg/day. Following these studies, choline acetyltransferase activity was significantly reduced in hippocampi extracted from the AF64A-treated rats, indicating successful cholinotoxicity. This paradigm may be useful as a possible screen for potential Alzheimer's disease therapeutic agents. This conclusion is supported by published reports of beneficial arecoline effects observed following 2-week intravenous infusions in patients with Alzheimer's disease (Soncrant, Raffaele, Asthana, Berardi, Morris, & Haxby, 1993). PMID- 9398594 TI - Long-term enhancement but not short-term in Hermissenda is dependent upon mRNA synthesis. AB - The reversible transcription inhibitor 5,6-dichloro-1-beta-D-ribobenzimidazole (DRB) was used to examine the contribution of mRNA synthesis to long-term enhancement (LTE) following one-trial conditioning of Hermissenda. Inhibition of mRNA synthesis by DRB or inhibition of protein synthesis by anisomycin did not significantly affect the induction and maintenance of short-term enhancement (STE) examined 1 h after one-trial conditioning. In contrast to the absence of an effect of the inhibitors on STE, LTE was blocked by DRB or anisomycin applied shortly before and during the presentation of the conditioning trial. Consistent with previous reports, animals that received an unpaired CS and US did not exhibit LTE. In addition, a control group that received a concentration of DRB (10(-7) M) that does not significantly affect mRNA synthesis exhibited typical LTE when tested 24 h postconditioning. These results demonstrate that the induction of LTE produced by one-trial conditioning is dependent upon transcription and the regulation of gene expression. PMID- 9398595 TI - Biochemical and Genetic Tests for Inhibitors of Leishmania Pteridine Pathways AB - The study of antifolate-resistant mutants of the protozoan parasite Leishmania has provided useful information about genetic processes such as gene amplification and mutation and knowledge of the unique features of the pteridine metabolic pathway in this primitive eukaryote. The novel bifunctional dihydrofolate reductase-thymidylate synthase (DHFR-TS) is an essential enzyme, yet most DHFR-TS inhibitors show little promise as potential drugs. Leishmania possess a novel alternative pteridine reductase (PTR1) which is relatively insensitive to methotrexate. We have proposed that the ability of PTR1 to serve as a metabolic bypass and thus modulate drug inhibition of DHFR-TS activity may be responsible for the poor efficacy of many antifolates. In this work, we have sought inhibitors of L. major PTR1 from a collection of 74 compounds. The most potent inhibitors were also tested against L. major DHFR-TS and human DHFR and several compounds showing good activity for PTR1 alone, or for all three reductases, were identified. The activity of these compounds was tested against wild-type promastigotes, and those which were potent inhibitors of both PTR1 and DHFR-TS (but not those active against only PTR1) showed good potencies. Growth inhibition tests of L. major mutants, lacking PTR1 or DHFR-TS (ptr1(-) and dhfr ts- knockouts) or overexpressing PTR1, were used as a "genetic screen" to assess whether these two pteridine reductases were targets in vivo. Remarkably, only one compound showed a methotrexate-like pattern of inhibition. Six compounds showed good inhibition of Leishmania growth regardless of PTR1 or DHFR-TS levels. These findings suggest that Leishmania cells contain multiple targets for a diverse set of antifolates, with one or more significant targets in addition to DHFR-TS and PTR1. This emphasizes the necessity of combined biochemical and genetic screens in efforts to rationally design chemotherapeutic strategies in Leishmania. Copyright 1997 Academic Press. Copyright 1997 Academic Press PMID- 9398596 TI - Tyrosine kinase signaling at fertilization. AB - The unfertilized egg is a highly differentiated cell that retains unlimited developmental potential. The execution of that potential requires signal transduction pathways that release the egg from its quiescent metabolic state, direct the union of the maternal and paternal genome, and initiate a developmental program that will guide embryogenesis. The egg is equipped with an array of cytosolic as well as cell surface receptor protein tyrosine kinases as part of a preassembled signal transduction mechanism. These protein tyrosine kinases have been found to act at several points during this egg activation process, beginning as early as the initial sperm-egg interaction. While many of these kinase functions are common to all cells, several functions unique to fertilization demonstrate the versatility of this class of protein kinases. PMID- 9398598 TI - Expression, purification, and characterization of a murine CD4 fragment containing the first two N-terminal domains. AB - CD4 is a membrane glycoprotein on T lymphocytes that binds to the same peptide:major histocompatibility complex (MHC) class II molecules recognized by the antigen-specific T cell receptor (TcR). Recent evidence supports the importance of coaggregation of CD4 and TcR for effective T cell activation. Here, we report that a transfected Chinese hamster ovary (CHO) cell line expressing a murine CD4 fragment containing the first two N-terminal domains secretes both monomeric molecules and disulfide-linked multimers. Elimination of the predicted N-linked glycosylation site at residue 161 that is next to the fourth cysteine does not affect the formation of interchain disulfide bonds. N-Terminal amino acid sequencing of the purified CD4 fragment demonstrates that the leader signal sequence is properly cleaved off the expressed protein. Circular dichroism studies suggest that both monomeric and disulfide-linked proteins are folded as primarily beta-sheet. PMID- 9398597 TI - Transcriptional analysis of the threonine dehydrogenase gene of Xanthomonas campestris. AB - The nucleotide sequence has previously been determined for the Xanthomonas campestris pv. campestris gene coding for threonine dehydrogenase (tdh). Flanking this gene are the upstream region possessing promoter activity and the downstream perfect inverted repeat having potential to form a stem-loop structure which resembles a transcription terminator. In addition, Northern blot analysis suggested the transcript of this gene to be monocistronic. In the present study, the essential region for promoter activity was narrowed down to a stretch of 57 bp which still retained 84% of the promoter activity. The first nucleotide to be transcribed is the guanosine at 30 nt upstream from the proposed tdh start codon. The putative terminator exhibited transcriptional termination activity bidirectionally in both Escherichia coli and X. campestris. These observations indicate that the transcriptional structure of X. campestris tdh is different from that of E. coli where tdh and kbl are organized into the tdh operon. Furthermore, the expression of tdh in X. campestris is repressed by leucine, a situation different from that in E. coli where leucine induces the expression of tdh operon. PMID- 9398599 TI - Recombinant Mycobacterium aurum expressing Escherichia coli beta-galactosidase in high throughput screening of antituberculosis drugs. AB - Mycobacterium aurum is considered a surrogate of M. tuberculosis and recently has been proposed as test organism in high throughput screening of antituberculosis drugs (3). In this investigation, we suggest use of a recombinant M. aurum expressing E. coli lacZ gene, in which beta-galactosidase production is the reporter system as recently reported by us (6). The assay is based on production of beta-galactosidase in presence of drugs during growth. Enzyme production was inhibited within 4 h by frontline antimycobacterial drugs like streptomycin, rifampicin, isoniazid, ethambutol, ofloxacin, and sparfloxacin at their MICs. The assay could be performed conveniently in 96-well microtiter plate format. PMID- 9398600 TI - A cryptic protease activity from Trypanosoma cruzi revealed by preincubation with kininogen at low temperatures. AB - Cysteine proteases have been identified in parasitic protozoa including the causative agent of Chagas' disease Trypanosoma cruzi. T. cruzi lysates subjected to substrate-containing SDS-polyacrylamide gel electrophoresis exhibit major bands of proteolytic activity in the 45-55 kDa molecular mass range (cruzipain activity). Paradoxically, addition of kininogen (a cystatin-like protease inhibitor) to the lysates before electrophoresis results in the appearance of additional bands of proteolytic activity in the 160-190 kDa molecular mass range. This inhibitor-activated protease activity depends upon the reaction conditions and exhibits novel properties. For example, a 24-48 hour preincubation at low temperature (-20 degrees C optimum) greatly enhances the proteolytic activity. The results suggest that a metastable complex forms between kininogen and a cryptic 30 kDa cysteine protease from T. cruzi and that this complex participates in the activation of proteolytic activity. PMID- 9398601 TI - Universal skew of T cell receptor (TCR) V beta usage for Crohn's disease (CrD). AB - It would be of clear interest and importance to identify T cell populations which correlate with the initiation of some T cell-mediated diseases; however, it is difficult to observe the initial response of T cells in these diseases because of modification due to immunosuppressive treatment. We investigated T cell receptor (TCR) V beta usage in both affected and unaffected mucosa from 16 patients with active Crohn's disease (CrD), undergoing nutritional therapy without any immunomodulatory medications. Semiquantitative reverse transcriptase-polymerase chain reaction showed increased expression of V beta 12 and 13 in the entire mucosa of CrD but not in the controls. This was confirmed by introducing a random cloning method. Such skewing was observed primarily in CD4+ lamina propria lymphocytes. DNA sequence analysis demonstrated a striking clonal expansion of V beta 12 T cells, but the dominant clones were not identical in the patients. These findings suggest the importance of superantigen as well as specific T cell response in the pathogenesis of CrD. PMID- 9398602 TI - Cardiac expressions of HIF-1 alpha and HLF/EPAS, two basic loop helix/PAS domain transcription factors involved in adaptative responses to hypoxic stresses. AB - Expression of many mammalian genes is regulated by oxygen tension. HIF-1 alpha and HLF/EPAS are two basic helix-loop-helix/PAS domain transcription factors that bind to hypoxia sensitive elements in the promoters /enhancers of hypoxia sensitive genes such as vascular endothelial growth factor (VEGF). This paper describes the structure of rat HIF-1 alpha and analyses expressions HIF-1 alpha and of HLF/EPAS mRNAs in lung and cardiac tissues from the rat. HLF/EPAS mRNAs appear at birth in the two tissues and are maintained at high levels throughout adult life. HIF-1 alpha mRNAs are expressed at a constant level during lung development. Their abundance increase transiently at birth in cardiac tissues. Cultured cardiomyocytes from new born rats only express HIF-1 alpha mRNAs. HIF-1 alpha mRNA expression is increased by phorbol myristate acetate but not by anoxia or cobalt. The results indicate (i) that HIF-1 alpha and HLF/EPAS are expressed in a cell specific manner and (ii) that the hypoxic induction of VEGF mRNA expression by isolated cardiomyocytes is independent of HLF/EPAS. Finally, they suggest that protein kinase C may prime hypoxia induced gene regulation by inducing expression of HIF-1 alpha mRNAs. PMID- 9398603 TI - New DNA minor-groove binding molecules with high sequence-selectivities and binding affinities. AB - New DNA minor-groove binding molecules with high binding affinities and sequence selectivities are described. The effects of structural changes in ligands with a three-ring backbone on their DNA-binding properties have been studied. Of a pool of eight potential ligands, two showed binding affinities and sequence selectivities rivaling distamycin. The two best ligands share a common structural motif that involves a para-di-substituted benzene ring flanked by two meta-di substituted benzene rings. PMID- 9398605 TI - Structural characterization of mouse monoclonal antibody 13-1 against a porphyrin derivative: identification of a disulfide bond in CDR-H3 of Mab 13-1. AB - The amino acid sequence of a mouse monoclonal antibody Mab13-1, a catalytic antibody against TCPP (meso-tetrakis(4-carboxyphenyl)porphyrin), was confirmed by mass spectrometric (MS) peptide mapping. The amino-terminal sequence of the heavy chain was established by MS/MS analysis of the isolated N-terminal peptide. The presence of a unique disulfide bond between Cys93H and Cys102H was identified by MS peptide mapping and sequence analysis of an S-S containing peptide. Positions of other disulfide bonds were identified to be conserved. The non-conserved disulfide bridge was found to be resistant as other intra-chain disulfide bonds against reduction under non-denaturing condition, and to be buried inside the molecule. This extra disulfide bond is expected to support antigen-binding by restricting the flexibility of CDR-H3 loop, and it might be favorable for the recognition of a plane antigen, a porphyrin derivative. PMID- 9398604 TI - Helicobacter pylori induces interleukin-8 expression in endothelial cells and the signal pathway is protein tyrosine kinase dependent. AB - Helicobacter pylori (HP) infection has been shown to increase gastric mucosal interleukin 8 (IL-8) expression, and whether HP or its toxin induces endothelial cell IL-8 expression is unknown. We aimed to compare the IL-8 expression in endothelial cells after stimulation with HP toxin, tumor necrosis factor alpha (TNF-alpha), and lipopolysaccharide (LPS) and to study their signal pathways. HP or its toxin induced significant IL-8 expression in endothelial cells. HP toxin, TNF-alpha, and LPS also showed a time- and dose-dependent increase in IL-8 expression over the control. Both protein kinase C (PKC) and protein kinase A (PKA) inhibitors had no effect on IL-8 response to these stimuli. Protein tyrosine kinase (PTK) inhibitor genistein at concentrations of 150, 300, and 450 microM dose-dependently reduced LPS- and TNF-alpha-induced IL-8 expression by 29.43, 43.8, and 47.3% and 20.5, 49.9, and 61.8% respectively, whereas HP toxin induced IL-8 secretion could only be reduced at 450 microM by 35.7%. Geldanamycin, a more potent PTK inhibitor, at doses of 0.5, 1, and 2 microM dose dependently reduced HP toxin induced endothelial cell IL-8 expression by 24.8, 26, and 44.3% respectively. It is concluded that HP and its toxin can increase IL 8 expression in endothelial cells, and the expression of IL-8 elicited by HP toxin, TNF-alpha, and LPS is partially dependent on PTK but not PKA or PKC activation. PMID- 9398606 TI - Heterogeneity of binding sites and bioeffects of raloxifene on the human leukemic cell line FLG 29.1. AB - The benzothiophene divarative raloxifene is known to mimic estrogen in human bone remodeling. To investigate the "in vitro" properties of raloxifene on osteoclast precursors, the human leukemic cell line FLG 29.1, which differentiates toward the osteoclastic phenotype, was examined for raloxifene binding and for evidence of its bioeffects. Scatchard and Hill analysis of binding data with the tritiated raloxifene demonstrated the presence of two classes of binding sites in both nuclear and cytosol fractions with Kd values of approximately 1 nM and approximately 5 nM, respectively. In addition, analysis of binding data using tritiated 17 beta E2 as ligand at high concentrations (10-40 nM) and either unlabeled 17 beta E2 or raloxifene as competitors gave similar results demonstrating the presence of type II EBS in these cells. Picomolar concentrations of raloxifene significantly (p < 0.05) inhibited cell proliferation. Moreover, the compound at nanomolar concentrations induced a significant dose- and time-dependent increase of progesterone receptor, and activated apoptotic cell death. These findings clearly demonstrate that raloxifene acts as an estrogen agonist in FLG 29.1 cells, acting through the estrogen receptor and, possibly, via multiple cooperative binding component(s). PMID- 9398607 TI - The rat S-adenosylhomocysteine hydrolase promoter. AB - The 1.2-kb DNA sequence flanking the transcription start of the AdoHcy hydrolase gene was cloned into the luciferase reporter plasmid pGL3-basic, and promoter activity was measured in transiently transfected CHO cells. Deletion analysis showed that most promoter activity was located within a 153 bp fragment immediately upstream from the predominant transcription start. The 153 bp fragment includes sites for AP-2, glucocorticoid-responsive element, SP-1, and a TATA-like sequence TATTTAAA. Mutational analysis demonstrated that the SP-1 site nearest the start of transcription contributed significantly to promoter activity, whereas, the other elements, including the appropriately positioned TATTTAAA sequence, had little affect on promoter activity. PMID- 9398608 TI - Expression and purification of His-tagged beta-1,4-galactosyltransferase in yeast and in COS cells. AB - His6-tag technology has been introduced for easy purification of recombinant proteins expressed in Escherichia coli. Aiming at extending this technology to purification of glycoproteins expressed in Saccharomyces cerevisiae or in animal cells, respectively, we adapted this protocol to recombinant soluble beta-1,4 galactosyltransferase (rgal-T). A His6-tag was introduced to the N-terminus of the protein (hisGal-T). The Histagged enzyme expressed in yeast S. cerevisiae was enzymically active but could not be purified from the cell extract by virtue of the His6-tag. Binding efficiency of hisGal-T was found to be impaired by a bulky N-glycan close to the His-tag. Removal of the unique site of N-glycosylation using site-directed mutagenesis restored binding of hisGal-T to the Ni-NTA resin. In comparison N-glycosylated hisGal-T transiently expressed in COS cells was secreted as a soluble active enzyme and could be purified in one single step by virtue of the His6-tag. PMID- 9398609 TI - In vitro lipolytic effect of leptin on mouse adipocytes: evidence for a possible autocrine/paracrine role of leptin. AB - The present study has examined the effects of the adipocyte-derived hormone, leptin, on lipolysis in fat cells of different types of mice. Exposure to leptin (1.25.10(-6) M to 1.25.10(-12) M) increased (P < 0.01) the lipolytic activity of fat cells obtained from lean mice. A greater stimulation was observed when adipocytes from ob/ob mice were examined. Throughout the concentrations tested, the leptin-induced lipolysis observed in fat cells of lean animals was smaller than that obtained in ob/ob mice. The maximal lipolytic effect in obese animals was observed with 10(-8) M of OB protein. The lipolytic activity following the addition of 1.25.10(-10) M to 1.25.10(-6) M was significantly increased (P < 0.01) in ob/ob mice compared to lean animals. Adipocytes from ob/ob mice responded in a dose-dependent manner to the OB protein, while the leptin-induced lipolysis observed in lean animals was dose-independent. In contrast to lean and ob/ob mice, leptin did not stimulate lipolysis in adipocytes from db/db mice, which have a mutation in the leptin receptor gene. These in vitro studies suggest an autocrine/paracrine action of leptin on white fat cells and envisages the involvement of the OB protein, not only in centrally mediated pathways, but also in physiological functions which take place peripherally. PMID- 9398610 TI - Glycated proteins modulate tissue-plasminogen activator-catalyzed plasminogen activation. AB - Plasminogen activation by tissue-plasminogen activator (t-PA) is accelerated by the presence of a macromolecular surface, which acts as a template that brings enzyme and substrate in close proximity. Modification of lysine residues, which are important for this template function, occurs in diabetic patients as a consequence of glycation of proteins. In this study, we investigated the effects of glycation of fibrin and other proteins in t-PA-catalyzed plasmin formation. Plasminogen activation on glycated fibrin(ogen) was increased compared to non glycated fibrin(ogen), which could fully be attributed to an increased affinity of t-PA for glycated fibrin(ogen). Binding of plasminogen to glycated fibrin was increased, but did not contribute to increased plasminogen activation. Both plasminogen activator inhibitor-1 (PAI-1) binding and activity were increased on glycated fibrin. Induction of template function in plasminogen activation was also observed on immobilized glycated bovine serum albumin (BSA) and human gamma globulins (IgG). Increased plasmin generation at sites of deposition of glycated proteins may lead to increased extracellular matrix breakdown and thereby affect the integrity of the endothelial monolayer. Moreover, soluble glycated BSA and glycated IgG can inhibit t-PA binding to immobilized glycated fibrin and interfere with fibrinolysis in diabetic patients. PMID- 9398611 TI - Phosphorylation of osteopontin by Golgi apparatus casein kinase. AB - Osteopontin (OPN) is a ubiquitous multiphosphorylated secretory glycoprotein. Twenty-seven phosphorylated serines have been identified in bovine milk OPN (E. S. Sorensen et al. (1995) Protein Sci. 4, 2040-2049). Nineteen of these phosphoacceptor sites are fully conserved in rat OPN, all displaying the consensus for the Golgi apparatus casein kinase, G-CK (S-x-E/Sp). Here we show that rat OPN is indeed phosphorylated more readily than casein itself by G-CK from either rat mammary gland or liver. OPN is also phosphorylated by casein kinases-1 and -2 (CK1, CK2), though less readily than casein. If OPN kinase activities are normalized in terms of casein phosphorylation, OPN phosphorylation rate by G-CK is 78-fold and 19-fold higher than those measured with CK2 and CK1, respectively. These data, in conjunction with the specific location of G-CK to the Golgi apparatus, where CK2 and CK1 are hardly detectable, support the view that G-CK is the main if not the only physiological agent committed to the phosphorylation of OPN. PMID- 9398612 TI - Human canalicular multispecific organic anion transporter (cMOAT) is expressed in human lung, gastric, and colorectal cancer cells. AB - Human canalicular multispecific organic anion transporter (cMOAT), a glutathione conjugate membrane transporter, has been isolated from cisplatin-resistant cancer cells and is distributed mainly in normal liver. We analyzed the expression of human cMOAT in 14 lung, 11 gastric, and 9 colorectal non-drug-selected human cancer cells, two multidrug-resistant cells, and one cisplatin-resistant cells, using quantitative RT-PCR and newly developed anti-human cMOAT antibody. All cell lines analyzed here expressed human cMOAT at the level of mRNA and protein, and some of them expressed higher levels of human cMOAT than the cisplatin-resistant cells. The two multidrug-resistant cell lines co-expressed human cMOAT gene and both or either of MRP and MDR1 genes. Immunostaining showed that human cMOAT was predominantly localized to the cytoplasm of these single cells. Our results indicate that human cMOAT is expressed in various human cancer cells including drug-resistant cells. PMID- 9398613 TI - Dihydropyridine receptor and ryanodine receptor gene expression in long-term denervated rat muscles. AB - Following disruption of the nerve supply, extensor digitorum longus (EDL) and soleus (SOL) muscles in rats are known to exhibit alterations in excitation contraction coupling. After total RNA isolation from the denervated and the contralateral control muscles performed at 25 and 50 days following denervation, RNase protection assays were carried out with four cDNA probes specific for the skeletal and cardiac isoforms of both the DHPR alpha 1-subunit and the RyR. Longterm denervation increased the expression of the mRNA for skeletal DHPR and skeletal RyR in SOL muscle, but it also significantly increased the expression of the mRNA for the cardiac isoform of the DHPR alpha 1 subunit in EDL muscle. PMID- 9398614 TI - Molecular assembly of the extracellular domain of P2X2, an ATP-gated ion channel. AB - We have produced the putative extracellular domain (ECD) of the ATP-gated ion channel, P2X2, in a bacterial expression system. The hexahistidine-tagged protein was purified by immobilized metal affinity chromatography and refolded by sulfitolysis and dialysis. We demonstrate that P2X2-ECD forms a stable tetramer in solution by gel filtration chromatography, dynamic light scattering and analytical sedimentation centrifugation. [alpha-32P]ATP has been covalently cross linked by UV irradiation to the P2X2-ECD and this binding is specific and competable by antagonists suramin and cibacron blue. These results indicate that the binding affinity among P2X2-ECD subunits is appreciably stronger than 3.4 microM (0.1 mg/ml), implying that the extracellular domain of P2X2 is primarly responsible for tetramerization of whole P2X2 and thus probably plays a role in determining homo- and heteromerization specificity of P2X channel subunits. PMID- 9398615 TI - Mutations and deletions within the S8-S9 interdomain region abolish complementation of N- and C-terminal domains of Ca(2+)-activated K+ (BK) channels. AB - A full length alpha-subunit of the Ca(2+)-activated K+ (BK) channel with an inactivating mutation in the C-terminus can complement a functional C-terminal fragment. We analyzed deletions and amino acid changes within the S8-S9 interdomain region for their ability to allow complementation. Cys612 and His616 that are located in a region that contains two overlapping signature sequences, a immunoglobulin signature sequence and a heme binding domain, are essential for a functional channel. These two amino acid residues are also essential for complementation. The deletion of the PEST sequence does not affect the function of the BK channel; however, without the PEST sequence, complementation by a functional C-terminal fragments is no longer possible. The ability to complement a functional channel is restricted to the C-terminal fragment and requires that the complete alpha-subunit or the larger N-terminal fragment contains both, the immunoglobulin signature sequence the PEST sequence. PMID- 9398616 TI - Bacterial lipopolysaccharide induces early and late activation of protein kinase C in inflammatory macrophages by selective activation of PKC-epsilon. AB - Experiments from our and other laboratories have shown that specific inhibitors of protein kinase C (PKC) inhibited the secretion of nitric oxide, TNF alpha, and IL-1 beta from lipopolysaccharide (LPS)-stimulated macrophages, suggesting an important role for PKC in the inflammatory response. The present study was designed to investigate the mechanism whereby LPS stimulates PKC activity in inflammatory macrophages. Mouse macrophages were stimulated with 0-1 microgram/ml LPS for 0-18 hours, and PKC activity was detected in cell lysates. PKC isoform specificity was determined by blocking PKC activity with isoform-specific antibodies. Treatment of macrophages with 1 microgram/ml LPS induced a two-fold increase in PKC activity within 15 minutes and an additional more significant peak of PKC activity appeared 3 hours post-LPS stimulation. A lower dose of LPS (10 ng/ml) induced the later peak only. The enhancement in PKC activity induced by LPS occurred in both the cytosol and membrane fractions, but the enhancement in the membrane fraction was significantly greater than in the cytosol. The increase in PKC activity in both peaks was abolished only by the addition of anti PKC-epsilon antibody. The present experiments suggest that PKC activation is an important pathway in the LPS-induced secretory response of macrophages and that PKC-epsilon is the major isoform involved. PMID- 9398617 TI - Interactions of FLT-1 and KDR with phospholipase C gamma: identification of the phosphotyrosine binding sites. AB - Vascular endothelial cell growth factor interacts with the receptor tyrosine kinases Flt-1 and KDR/Flk-1. We report that both receptors bind to PLC gamma and display specificity for the N-SH2 over the C-SH2 domain. Extensive site-directed mutagenesis of Flt-1 reveals that the juxta-membrane Y794, and the carboxyl terminal Y1169, are two major sites of interaction. Amino acids in the +1, +2 and +3 positions following these tyrosines are LSI and IPI, respectively. Peptide maps generated from wild type and mutant Flt-1 confirms that these residues are autophosphorylated. As predicted, mutagenesis of the analogous amino acids in KDR, positions Y801F and Y1175F, which lie in contexts YLSI and YIVL, respectively, reduced interactions of PLC gamma with this receptor. We conclude that both Flt-1 and KDR have the potential to signal through PLC gamma via phosphotyrosine residues located in juxta-membrane and carboxyl tail regions. PMID- 9398618 TI - Cell-cycle regulation of DNA damage-induced expression of the suppressor gene PML. AB - The promyelocytic leukemia (PML) gene, which encodes a growth- and transformation suppressor, has been identified at the non-random chromosomal translocation break point t(15; 17)(q22; q12) of acute promyelocytic leukemia. To elucidate if PML may play a role in cellular response to DNA damage, PML expression was analyzed by immunofluorescence staining in HeLa cells treated with ionizing radiation (IR) and cisplatin. Our studies demonstrated IR at 20Gy, and cisplatin at 6 micrograms/ml caused more than 5-10 fold increases in PML protein expression in the PML Oncogenic Domain (POD) by immunofluorescent staining. Northern blotting showed that there was no gross increase in mRNA levels indicating that the induction is a post-transcriptional event. Flow cytometry showed that HeLa cells treated with IR were progressively arrested in G1, which correlates with the optimal expression of PML in the cell cycle. To determine if PML expression was under the control of the tumor suppressor p53, which is known to arrest cells in G1, HeLa cells were transfected with the wild-type p53 gene. PML expression in p53 transduced cells were 5-10 fold higher than the control, indicating that the enhanced expression of PML is apparently dependent on the p53 pathway. These data also indicate that PML may play an important role in cellular response to DNA damage such as DNA repair or apoptosis during G1 arrest. PMID- 9398619 TI - Channel formation and [Ca2+]i accumulation induced by perforin N-terminus peptides: comparison with purified perforin and whole lytic granules. AB - Cytotoxic T lymphocytes (CTL) and natural killer (NK) cells express the pore forming protein perforin, which contributes to lymphocytotoxicity. The hallmark of perforin action is opening high-conductance transmembrane channels that enable massive influx of Ca2+ ions (deleterious to many cell types), as well as granzymes, which may trigger the apoptotic pathway. To explore the functional domains in the perforin molecule, we investigated in PN71 lymphocytes, the ability of perforin N-terminus synthetic peptides (compared to purified perforin and perforin-containing lytic granules), to cause intracellular Ca2+ ([Ca2+]i) accumulation and open transmembrane channels. To this end, we used the whole cell recording technique and Indo 1 fluorescence to measure membrane currents and [Ca2+]i, respectively. We have demonstrated that the N-terminus peptide Hu-34 (amino acids 1-34) closely resembled perforin action, reflected by [Ca2+]i accumulation and channel activity, while shorter peptides (e.g., Hu-16) generated mostly short-lived channels but no [Ca2+]i elevation. Hence, the first 34 amino acids of the perforin N-terminus sequence are sufficient for the perforin action. PMID- 9398620 TI - TTX resistivity of Na+ channel in newt retinal neuron. AB - We examined voltage-dependent, TTX-resistant Na+ channels of newt retina (nRNaCh) electrophysiologically. IC50-TTX value of nRNaCh is more than 20 microM. We determined partial cDNA sequences of nRNaCh restricted to TTX binding sites (SS2 regions of all four repeats). While the amino acid sequences of SS2 regions of repeats II, III and IV of nRNaCh are identical to those of TTX-sensitive Na+ channels, only one amino acid in SS2 of repeat I of nRNaCh is different. nRNaCh have nonaromatic amino acid (Ala) in this site instead of the aromatic amino acid in a case of TTX-sensitive Na+ channels. Many studies suggested that the differences of TTX sensitivity of Na+ channels are decided by whether amino acid in this site is aromatic or not. Therefore nRNaCh acquire their TTX resistivity with the same mechanism as TTX-resistant cardiac Na+ channels do. PMID- 9398621 TI - Presence of antibacterial peptides on the laid egg chorion of the medfly Ceratitis capitata. AB - Female reproductive accessory glands of the medfly Ceratitis capitata produce a secretion with antibacterial activity mainly ascribed to ceratotoxin peptides. To study whether the secretion from the accessory glands of the female protects the eggs and early larva from microbes, we examined whether ceratotoxins and other accessory gland components could be found on the egg surface. This was found to be the case; a water-soluble material with the same protein and antibacterial pattern as that of the accessory gland secretion was recovered from the laid egg surface and was observed as electrondense, clustered droplets over the outer exochorion. Such material showed the same electrophoretic pattern in both mated and virgin females. These findings indicate that the accessory gland secretion is spread, at oviposition, onto the eggs producing an antibacterial coating, irrespective of fertilization. This is the first report of antimicrobial components recovered from a material layered on insect laid eggs. PMID- 9398622 TI - Nitric oxide is not involved in lipopolysaccharide and cytokine mixture-induced cellular injuries in primary cultured hepatocytes. AB - Nitric oxide (NO) from artificial NO donors induces cell death through complete inhibition of mitochondrial respiration of hepatocytes. Treatment of hepatocytes with lipopolysaccharide (LPS) and cytokine mixture (interferon gamma and tumor necrosis factor alpha) not only results in NO production but also causes cellular respiration suppression and cell death in hepatocytes. NG-monomethyl-L-arginine, a specific inhibitor of nitric oxide synthase, inhibits hepatocyte NO synthesis but cannot prevent hepatocytes from LPS and cytokine mixture-induced cellular injuries. Similarly, some metabolic intermediates capable of inhibiting hepatocyte NO synthesis cannot block LPS and cytokine mixture-mediated cellular injuries in hepatocytes. These results imply that lipopolysaccharide and cytokine mixture-induced cellular injuries and NO syntheses are parallel events, NO is not involved in LPS and cytokine mixture-induced cell damage. PMID- 9398623 TI - Evidence that exposure of particulate air pollutants to human and rat alveolar macrophages leads to differential oxidative response. AB - Macrophages and inflammatory cells generate active oxygen species in the process of killing and degrading microorganisms. Air pollutant particles may be ingested by macrophages and stimulate the same mechanisms to produce a long term oxidative burden to the lung if particles are not degraded. In the present study human and rat alveolar macrophages (AM) were compared in their response to inhaled particles using luminol dependent chemiluminescence (CL) and peroxide dependent CL assays. Cytotoxicity was measured by the release of lactate dehydrogenase (LDH) activity in the supernatant. Human AM produced more oxidants than rat AM whether, unstimulated, after addition of particles or addition of particles then peroxidase. Human AM also had a different spectrum of response to the same particles. Our results suggest that human macrophages produce more reactive oxygen species in respond to particles than rat AM. PMID- 9398624 TI - Binding specificity of avian heat shock protein 108. AB - Chicken heat shock protein 108 (HSP108), the avian homolog of GRP94, was originally isolated from hen oviduct and binds Fe-ovotransferrin (Fe-OTf). The liver is also a rich source, and liver membranes bind Fe-OTf with a KD of 1.7 x 10(-7) M, a value similar to oviduct membranes. A competition assay, based on the binding of 125I-Fe-OTf to liver membranes, was utilized to examine the binding specificity of HSP108. Ovalbumin and avidin competed effectively, with KD's of 1.8 x 10(-7) M and 1.4 x 10(-7) M, respectively. Iron-free OTf bound with a 10 fold higher KD. Egg white lysozyme, chicken IgG, human transferrin, rabbit muscle actin, and porcine insulin do not bind. Neither do denatured ovalbumin or ovalbumin tryptic peptides. Thus, the binding activity of HSP108 is not restricted to Fe-OTf, nor is it universal. PMID- 9398625 TI - Differences in hypervariable region 1 quasispecies between immune complexed and non-immune complexed hepatitis C virus particles. AB - Antibody to the hypervariable region 1 of hepatitis C virus (HCV) is thought to have neutralizing activity. The complexity of hypervariable region 1 quasispecies was compared between immune complexed and non-immune complexed HCV particles. Immune complexes and non-immune complexes, including intact HCV virions, were separated by differential flotation centrifugation and immunoprecipitation, and immune complexes were observed in 9 of 11 patients with chronic hepatitis C. Considerable differences in both hypervariable region 1 quasispecies and predominant clones were demonstrated between immune complexes and non-immune complexes in 4 patients and not in 5 patients. These results suggest that the specificity of antibody to hypervariable region 1 is related to the formation of immune complexes and that escape mutants with highly different quasispecies are resistant to neutralization by antibody to hypervariable region 1. PMID- 9398626 TI - Downregulation of MAP kinase activity signalled by HIV-1-gp120 coat protein in granular neurons and glial cells from rat cerebellum. AB - We have studied the effect of gp120 coat protein from HIV-1 on tyrosine phosphorylation processes in primary cultures of granular neurons or glial cells from the cerebellum of neonatal rats. The extracellular application of recombinant gp120 (200 pM) was able to reduce the phosphotyrosine content and the immunoreactivity for active form-specific antibodies of MAP kinase. Whereas in neurons MAP kinase appeared to be the only protein whose phosphotyrosine content was decreased, in glial cultures the inhibitory effect of gp120 on tyrosine phosphorylation processes appeared to be more widespread. In neuronal cultures, the effect of the viral protein was prevented by the concomitant treatment with depolarizing agents. PMID- 9398627 TI - Aberrant tau phosphorylation and neurite retraction during NGF deprivation in PC12 cells. AB - Recently apoptotic markers have been found in Alzheimer's Disease (AD) brain. To investigate the relation between tau phosphorylation and apoptosis, immunocytochemistry of AT8 (indicating the degree of phosphorylation at the tau Ser202/Thr205 site) was quantitatively determined the degree of tau phosphorylation at the Ser202 site was monitored during neuronal apoptosis in differentiated PC12 cells after nerve growth factor (NGF) deprivation. During this programmed cell death a prominent retraction of neurites took place that was associated with a clear increase in the level of AT8 signalaberrant phosphorylated tau at the Ser202 site. The broad spectrum kinase inhibitor staurosporine attenuated both this increase in tau phosphorylation, neurite retraction, and apoptosis. We suggest that at some point during programmed cell death, kinases with tau as substrate become activated and that the resulting loss of cytoskeletal integrity leads to neurite instability. PMID- 9398628 TI - Wortmannin, a specific inhibitor of phosphatidylinositol-3-kinase, enhances LPS induced NO production from murine peritoneal macrophages. AB - To elucidate the role of phosphatidylinositol-3-kinase (PI3K) during macrophage activation, we examined the effects of wortmannin, a specific inhibitor of PI3K, on the induction of nitric oxide (NO) synthesis and tumor necrosis factor-alpha (TNF-alpha) secretion from lipopolysaccharide (LPS)-stimulated macrophages. Wortmannin had no effects on NO synthesis and TNF-alpha secretion by itself. Wortmannin markedly potentiated the LPS-induced NO production in a dose-dependent manner. Western blot analysis demonstrated that significantly increased levels of iNOS protein were expressed in LPS-stimulated macrophages treated with wortmannin, compared to those without LPS. Furthermore, enhancement of TNF-alpha secretion was observed in the initiation stage for activation of LPS-stimulated macrophages treated with wortmannin. These results suggest that PI3K plays an important role in transducing the signal that is involved in LPS-induced macrophage activation. PMID- 9398629 TI - Nuclear accumulation of G-actin in isolated rat hepatocytes by adenine nucleotides. AB - Extracellular ATP induces bleb formation in isolated rat hepatocytes. We examined the effect of extracellular ATP on the actin cytoskeleton of these hepatocytes. Exposure to 100 microM ATP caused pronounced nuclear accumulation of G-actin. ADP, AMP, adenosine, and dibutyryl-cAMP induced the same effect. Adenosine deaminase could inhibit both ATP- and adenosine-induced nuclear accumulation. The P2-receptor agonists, UTP and 2' & 3'-O-(4-benzoylbenzoyl)-adenosine 5' triphosphate, did not induce this redistribution of G-actin. Phalloidin, which prevents depolymerisation of F-actin filaments to G-actin monomers, inhibited adenosine-induced nuclear accumulation of G-actin. These observations suggest that nuclear accumulation of G-actin is mediated by adenosine receptors. PMID- 9398630 TI - A conserved region in the first intron of the insulin receptor gene binds nuclear proteins during adipocyte differentiation. AB - The insulin receptor gene is induced 8 to 10-fold during adipocyte differentiation. Plasmids containing the promoter, exon 1 and a portion of the first intron from either the mouse or human gene are able to modulate the expression of an insulin receptor/CAT gene 3 to 7-fold during differentiation. We have shown that several nuclear proteins from both preadipocyte and adipocyte nuclear extracts bind to two discrete sites within a 278-bp region in the 5' end of the first intron. Sequence comparison between the first intron of the human gene and the mouse gene shows two regions of sequence identity which correspond to the protein binding regions detected by DNase footprinting. One of these sites binds proteins that are enriched in adipocyte nuclear extracts and can be competed by adipose regulatory element, ARE6. PMID- 9398631 TI - Insulin-like growth factor binding proteins-3 and -5 form sodium dodecyl sulfate stable multimers. AB - Insulin-like growth factor binding proteins (IGFBPs) are important modulators of IGF actions. IGFBP-3 and IGFBP-5 can bind to the extracellular matrix of a number of cell types. We now describe a new posttranslational structural modification of IGFBP-3 and IGFBP-5, which could play a role in determining their localization. We incubated radioiodinated forms of all six IGFBPs in the presence of a redox buffer consisting of 10 mM reduced glutathione and 0.2 mM oxidized glutathione. Under these conditions IGFBP-3 and IGFBP-5, but not the other IGFBPs, formed high molecular weight disulfide-linked multimers. Heparin and a peptide encompassing the high-affinity heparin-binding site in the C-terminal portion of IGFBP-3 were capable of blocking the multimerization of IGFBP-3. IGFBP-3, but not IGFBP-1, was shown to be able to self-associate non-covalently, which could be a requisite first step in the formation of covalent multimers. The self-association of IGFBP 3 required the high-affinity heparin-binding site in the C-terminal portion of the molecule. PMID- 9398632 TI - Detection of apolipoprotein E/dimeric soluble amyloid beta complexes in Alzheimer's disease brain supernatants. AB - The inheritance of the apolipoprotein (apo) E4 allele is an important risk factor for late-onset Alzheimer's disease (AD). A major component of the Alzheimer's disease neuritic plaques is amyloid beta (A beta). We previously identified apoE/A beta complexes within neuritic plaques (1). It was not known if this interaction takes place before or after A beta peptides become incorporated into neuritic plaques. To address this question we sought evidence of apoE complexes with brain soluble A beta peptides in AD and control patients. In addition, numerous proteins have been shown to bind A beta peptides in vitro. It is not know if any of these bind brain sA beta in vivo. We found evidence for the presence of apoE/dimeric sA beta complexes in the AD brain and could not detect complexes with other A beta peptide binding proteins. The binding of sA beta to apoE may be one factor influencing its clearance from the brain and/or its conformational state. PMID- 9398633 TI - Oligomerization is an intrinsic property of calsequestrin in normal and transformed skeletal muscle. AB - In skeletal muscle fibers, the high-capacity medium-affinity Ca(2+)-binding protein calsequestrin functions as the major Ca(2+)-reservoir of the sarcoplasmic reticulum. To determine the oligomeric status of calsequestrin, immunoblotting of microsomal proteins following chemical crosslinking was performed. Diagonal non reducing/reducing two-dimensional gel electrophoresis was employed to unequivocally differentiate between cross-linked species of 63 kDa calsequestrin and calsequestrin-like proteins of higher relative molecular mass. Since chronic low-frequency stimulation has a profound effect on the expression of many muscle specific protein isoforms, we investigated normal and conditioned muscle fibers. Calsequestrin was found to exist in a wide range of high-molecular-mass clusters in normal and chronically stimulated skeletal muscle fibers. Hence, oligomerization is an intrinsic property of this important Ca(2+)-binding protein and does not appear to be influenced by the fast-to-slow transformation process. Although fiber-type specific differences exist in the physiology of the skeletal muscle Ca(2+)-regulatory system, oligomerization of calsequestrin seems to be essential for proper functioning. PMID- 9398634 TI - Determination of a cleavage site of presenilin 2 protein in stably transfected SH SY5Y human neuroblastoma cell lines. AB - Mutations in the presenilin 1 (PS1) and presenilin 2 (PS2) genes are associated with early-onset autosomal dominant familial Alzheimer's disease, and the gene products are endoproteolytically processed to yield N-terminal fragments (NTF) and C-terminal fragments (CTF). We have studied the cleavage site of the PS2 protein in stably transfected human neuroblastoma cells. The 23 kD PS2-CTF was isolated by a combination of anion exchange chromatography and affinity chromatography and directly sequenced. The N-terminus of the PS2-CTF started at residue 307, which indicated that the cleavage occurs between Lys306 and Leu307 in the proximal portion of the large hydrophilic loop. This site is close to the cleavage positions observed in the PS1 protein. PMID- 9398635 TI - Ursodeoxycholic acid enhances glucocorticoid-induced tyrosine aminotransferase gene expression in cultured rat hepatocytes. AB - Ursodeoxycholic acid (UDCA) is an effective treatment for immune-mediated liver diseases, suggesting that UDCA is functionally similar to glucocorticoids (GCs). We investigated the effects of UDCA on the enzyme activity and the mRNA levels of tyrosine aminotransferase (TAT), a hepatocyte-specific marker of GC action, in primary cultured rat hepatocytes. Addition of UDCA resulted in a significant increase in TAT activity in the presence of dexamethasone (DEX), compared with DEX alone, and this increase was completely suppressed by sphingosine, a protein kinase C (PKC) inhibitor, or actinomycin D, a transcriptional inhibitor. UDCA could not induce TAT activity in the absence of DEX. UDCA increased the TAT mRNA levels in the presence of DEX. In conclusion, UDCA enhances the GC-induced TAT gene expression in hepatocytes, and UDCA-activated PKC may play a role in this upregulation. PMID- 9398636 TI - Cloning of cDNAs encoding G protein-coupled receptor expressed in human endothelial cells exposed to fluid shear stress. AB - A cDNA library of human umbilical vein endothelial cells exposed to fluid shear stress was constructed to search for functional endothelial genes expressed under flow conditions, and cDNAs encoding members of the G protein-coupled receptor (GPCR) family were cloned by a polymerase chain reaction (PCR) method using degenerate oligonucleotide primers. One of the two GPCR clones obtained was edg 1, and the other clone is a novel gene named FEG-1 that encodes a 375-amino acid protein similar to the receptors for both angiotensin II and chemokines. Reverse transcriptase-PCR showed that the FEG-1 and edg-1 mRNA levels in endothelial cells increased markedly in response to fluid flow. This suggests that FEG-1 and edg-1 may be receptor genes that play important roles in the regulation of endothelial function under physiological blood flow conditions. PMID- 9398637 TI - Leptin is present in human milk and is related to maternal plasma leptin concentration and adiposity. AB - Leptin is elevated during pregnancy and may be involved in the regulation of milk production in women. Immunoreactive leptin was quantified in human milk by modified radioimmunoassay. Leptin concentration was higher in whole vs. skim milk fractions; however, leptin concentration was not correlated with percentage milk fat. Leptin concentrations in whole and skim milk were correlated with maternal plasma leptin concentrations, maternal body weight, body mass index, and tricep skinfold thickness, but not with plasma insulin concentration. These data provide the first evidence for the presence of leptin in human milk in the range of concentrations found in human plasma and indicate that the concentration of leptin in milk reflects maternal adiposity. Determining the biological role(s) of milk-borne leptin could add to our understanding of neonatal metabolism and the mechanisms underlying the development of body fat and obesity in humans. PMID- 9398639 TI - Hydroxy juvenile hormones: new putative juvenile hormones biosynthesized by locust corpora allata in vitro. AB - The in vitro production of sesquiterpenoids was investigated by using corpora allata (CA) of the African locust Locusta migratoria migratorioides. Labeled products from unstimulated biosynthesis were extracted, purified by normal phase HPLC, and derivatized to determine the functional groups present. An extra hydroxyl group was detected in each of two juvenile hormone (JH) biosynthetic products. One compound, NP-8, was found to co-migrate with a chemically synthesized (Z)-hydroxymethyl isomer, 12'-OH JH-III, but not with the (E) hydroxymethyl isomer, 12-OH JH III. Mass spectral analyses further supported the identity of the synthetic material with that biosynthesized by the corpora allata. A second compound was identified as the 8'-OH JH-III based on spectroscopic analyses. 12'-OH JH-III exhibited morphogenetic activity when tested on the heterospecific Tenebrio test. These data suggest that 12'-OH JH-III and 8'-OH JH-III are additional biosynthetically-produced and biologically-active juvenile hormones, and constitute the first known members of the class of hydroxy juvenile hormones (HJHs). PMID- 9398640 TI - APP gene promoter constructs are preferentially expressed in the CNS and testis of transgenic mice. AB - Transgenic animals were used to examine the spatial and temporal regulation of the human beta amyloid precursor protein (APP) gene promoter region in vivo. A 2.9 kb DNA fragment encompassing the APP gene promoter was fused to the chloramphenical acetyltransferase (CAT) reporter gene (pAMY-CAT) or a partial cDNA encoding the potentially amyloidogenic C-terminal 100 amino acid region of APP (pAMY-C100). Expression of these transgenes occurred primarily, but not exclusively, in the central nervous system (CNS) and testis in multiple independent lineages of transgenic mice. Temporal expression of the CAT reporter gene during development paralleled that reported for the endogenous APP gene. These studies suggest that a CNS-responsive cis-acting element(s) may exist in the promoter/5'-flanking region of the APP gene. PMID- 9398641 TI - Receptor-mediated calcium entry is required for maximal effects of platelet activating factor primed responses in human neutrophils. AB - The effect of receptor mediated calcium entry (RMCE) on platelet activating factor (PAF) stimulated human neutrophils (PMNs) was investigated using SKF 96365, a selective inhibitor of RMCE. Changes in cytosolic calcium concentration were determined by the fluorometric dye indo-1, AM, superoxide generation by cytochrome c reduction, and CD11b surface expression by flow cytometry. SKF 96365 pre-treatment diminished the cytosolic calcium rise in PAF primed PMNs. SKF 96365 treatment significantly (p < 0.05) decreased superoxide generation in PAF primed PMNs, but did not affect activation of the PMN oxidase by fMLP or PMA. Chelation of extracellular calcium by EGTA, intracellular calcium by BAPTA, AM, and RMCE blockade by SKF 96365 all statistically inhibited the PAF induced increase in surface expression of CD11b (p < 0.05); moreover, SKF 96365 inhibited to a greater extent than EGTA or BAPTA, AM treatment. These results suggest that RMCE is required for maximal effects of PAF on PMN function. PMID- 9398642 TI - A novel brain gene, norbin, induced by treatment of tetraethylammonium in rat hippocampal slice and accompanied with neurite-outgrowth in neuro 2a cells. AB - Tetraethylammonium (TEA) induces long-term potentiation (LTP)-like synaptic enhancement in rat hippocampal slices. To find the genes related to this phenomenon, subtraction screening was performed between the mRNA of TEA-treated slices and that of untreated whole brain. One of the clones induced by the TEA treatment, named as norbin, was expressed only in neural tissues. The predicted protein sequence of norbin consisted of 729 amino acids, and no homologies in the sequence were found with known genes or proteins. Overexpression of norbin in cultured Neuro 2a cells by cDNA transfection induced neurite-outgrowth. Since in the course of neural plasticity the formation of new synapses should occur, the neurite-outgrowth-related protein, norbin, might play an important role in neural plasticity. PMID- 9398643 TI - Potentiation of depolarization-induced calcium release from skeletal muscle triads by cyclic ADP-ribose and inositol 1,4,5-trisphosphate. AB - Two second messengers, cyclic ADP-ribose (cADPR) and inositol 1,4,5-trisphosphate (IP3), potentiated the Ca2+ release from sarcoplasmic reticulum induced by transverse tubular membrane depolarization monitored in a triadic vesicle prepared from skeletal muscle. However, without depolarization they could not trigger the Ca2+ release. On the contrary, only cADPR potentiated caffeine induced Ca2+ release. Because Ca2+ releases potentiated by cADPR and IP3 were inhibited by 1 microM ruthenium red and 100 microM ryanodine, probably these second messengers potentiated the Ca2+ release through ryanodine receptor Ca2+ channels. These results suggest that in skeletal excitation-contraction coupling, cADPR and IP3 play a role as a potentiator or a modifier in vivo, but both modification pathways are different from each other. PMID- 9398644 TI - Spermine conjugated peptide nucleic acids (spPNA): UV and fluorescence studies of PNA-DNA hybrids with improved stability. AB - Peptide Nucleic Acids (PNAs), the achiral DNA mimics with amide backbone, are emerging as attractive leads for drug development by antisense approach. Two major limitations of PNAs from an application perspective are their limited solubility in aqueous systems and pronounced self-organization. In this paper, it is shown that covalent conjugation of spermine at C-terminus of PNA (spPNA) improves its solubility and binds to complementary DNA 20 times stronger than the corresponding binding of PNA. Fluorescence kinetics shows a 2 fold acceleration of the bimolecular association process in spPNA:DNA hybrids, due to electrostatic interaction cationic spermine tagged to PNA with anionic DNA. This modification is easy to incorporate into PNA synthetic protocols to make them more effective in biological applications and may improve the poor cell uptake of PNA. PMID- 9398645 TI - Single transduction in the transcriptional activator CooA containing a heme-based CO sensor: isolation of a dominant positive mutant which is active as the transcriptional activator even in the absence of CO. AB - We constructed an in vivo reporter system to measure the activity of CooA as the transcriptional activator and showed that the recombinant CooA was active as the transcriptional activator in the presence of CO even in E. coli cells. A dominant positive mutant of CooA, in which Met131 was replaced by Leu, was isolated by a random mutagenesis with this reporter system. The electronic absorption spectra of M131L mutant were identical to those of wild type CooA in oxidized (Fe3+), reduced (Fe2+), and CO-bound (CO-Fe2+) state, indicating that the coordination structure and environment of the heme were not changed by this mutation. Methionine at position 131 was the carboxyl-terminal end of the heme-binding domain of CooA, which would be adjacent to the hinge region connecting the heme binding domain and the DNA-binding domain. PMID- 9398646 TI - A pluripotent polyphenol oxidase from the melanogenic marine Alteromonas sp shares catalytic capabilities of tyrosinases and laccases. AB - The recently characterized marine melanogenic bacterium MMB-1 contains a pluripotent polyphenol oxidase (PPO) which catalyzes the oxidation of a very wide range of substrates considered specific for tyrosinase or laccase. This range includes monophenols such as L-tyrosine, o-diphenols such as L-dopa, p-diphenols such as hydroquinone, o-aminophenols such as 3-hydroxyanthranilic acid, activated monophenols such as 2,6-dimethoxyphenol and syringaldazine, and chromophores such as ABTS. This is the first report of an enzyme that is able to catalyze the oxidation of compounds so far considered specific for tyrosinases (L-tyrosine) or laccase (syringaldazine), showing cresolase, catechol oxidase and laccase activities. Such PPO could be a very useful model to study the structural requirements, catalytic mechanisms and involvement of the copper sites existing in non-blue and blue copper-oxidases. PMID- 9398647 TI - Positively charged liposome functions as an efficient immunoadjuvant in inducing immune responses to soluble proteins. AB - To design an optimum liposome immunoadjuvant for soluble protein antigens, we investigated the relationship between liposomal surface charge and adjuvant action. Positively charged multilamellar vesicles (MLV) were taken up efficiently by macrophages, while negatively charged and neutral MLVs were hardly picked up. Consistent with this, positively charged MLVs containing soluble chicken egg albumin (OVA) functioned as a more potent inducer of antigen-specific cytotoxic T lymphocyte (CTL) responses and antibody production than negatively charged and neutral MLVs containing the same concentrations of antigens. These results indicate that the positive charge on the surface of liposomes represents an important factor for enhancing their immunoadjuvancy in the induction of antigen specific immune responses. PMID- 9398648 TI - The placenta is not the main source of leptin production in pregnant rat: gestational profile of leptin in plasma and adipose tissues. AB - The gestational profiles of leptin in plasma, adipose tissue and placenta were investigated in rats. Leptin in plasma and the adipose tissue of the maternal compartment increased as pregnancy advanced and remained high during the latter half of pregnancy (approximately 2-fold compared with the non-pregnant state), followed by a rapid decrease just before parturition. Leptin in fetal plasma and amniotic fluid was first detectable on day 19 and then increased to levels comparable to those in maternal plasma of non-pregnant and day 21 pregnant rats. The total amount of mRNA in maternal adipose tissues significantly increased as pregnancy advanced and reached about 2.5-fold of those of non-pregnant rats on day 19 of pregnancy, followed by a marked decrease on day 3 of lactation. Placentae and decidual tissues did not show any expression of leptin mRNA on day 12 and 19 of pregnancy. These results indicate that the placenta is not a major source of leptin production in rats and also suggest the physiological significance of leptin in rat pregnancy. PMID- 9398649 TI - Interaction of mutagenic tryptophan pyrolysate with d(CGATCG)2: intercalation model based on NMR experiments. AB - The solution structure of the complex of 3-amino-1,4-dimethyl-5H-pyrido[4,3 b]indole (Trp-P-1), a potent mutacarcinogen isolated from tryptophan pyrolysate, with the hexamer duplex d(CGATCG)2, was analysed by 1H-NMR spectroscopy and molecular dynamic calculation. Trp-P-1 was intercalated between the CpG base pairings in a suitable manner to form the guanosine-Trp-P-1 adduct which corresponds to the major reactant of Trp-P-1 with DNA. PMID- 9398650 TI - Intronic sequences are involved in neural targeting of human dopamine transporter gene expression. AB - Dopamine transporter (DAT) plays a key role in terminating synaptic dopaminergic transmission. DAT acts exclusively on the plasma membrane of presynaptic dopaminergic neurons and DAT gene is an appropriate model for the study of dopaminergic neuron-specific regulation of gene activity. DAT represents an important target for widely used neuroleptic drugs and psychostimulants and for catecholamine-selective neurotoxins. Functional abnormalities of DAT have been implicated in diverse neurologic and psychiatric disorders. Understanding the mechanisms regulating human DAT gene activity is an important step towards elucidation of the molecular bases of a number of disorders and psychostimulant drug abuse and dependence. In this study we have cloned and characterised a 7-kb segment of the human DAT gene which includes at least 4 kb of its 5'-flanking region, localised its essential, or core-promoter, and identified the region involved in regulation of DAT neurospecific expression. PMID- 9398651 TI - CD36 forms covalently associated dimers and multimers in platelets and transfected COS-7 cells. AB - CD36 is a transmembrane glycoprotein expressed on the surface of a number of cell types. The analysis of CD36 from platelets using immunoblotting, gel filtration, and native PAGE indicated the presence of high molecular complexes exceeding the Mr of monomeric CD36. Experiments using transfected COS-7 cells revealed these complexes were homodimers and -multimers of CD36. The multimers could be dissociated by treatment with a reducing agent, indicating they were formed by intermolecular cysteine-bridging. Mutagenesis of the cDNA for CD36 implicated the cysteines in the extracellular domain of the molecule. The potential physiological roles of CD36 multimerisation are discussed. PMID- 9398652 TI - The lipoic acid analogue 1,2-diselenolane-3-pentanoic acid protects human low density lipoprotein against oxidative modification mediated by copper ion. AB - 1,2-Diselenolane-3-pentanoic acid, in which the sulfur atoms of alpha-lipoic acid are replaced with selenium, displayed markedly different antioxidant properties when compared to alpha-lipoic acid. 1,2-Diselenolane-3-pentanoic acid was unable to inhibit protein oxidative modification of human low density lipoprotein (LDL) and bovine serum albumin induced by copper ion or hydroxyl radical, whereas alpha lipoic acid showed significant protection. However, 1,2-diselenolane-3-pentanoic acid was able to inhibit the formation of lipid peroxidation products in LDL after oxidation by copper, while alpha-lipoic acid did not. Hence the diselenium compound exerts its effects in a lipophilic environment whilst lipoic acid exerts its effects in a hydrophilic environment. These differences in antioxidant activities of the two compounds may be explained, at least in part, by their differing partition coefficients. PMID- 9398653 TI - Modulation of translation initiation in rat skeletal muscle and liver in response to food intake. AB - Protein synthesis is altered in both skeletal muscle and liver in response to nutritional status with food deprivation being associated with an inhibition of mRNA translation. In the present study, the effect of food-intake on the initiation of mRNA translation was examined in rats fasted for 18-h and then refed a complete diet. Fasting and refeeding caused alterations in translation initiation in both skeletal muscle and liver that were not associated with any detectable changes in the activity of eIF2B or in the phosphorylation state of eIF2 alpha. Instead, alterations in initiation were associated with changes in the phosphorylation state of eIF4E and/or the association of eIF4E with eIF4G as well as the eIF4E binding protein, 4E-BP1. In muscle from fasted rats, the amount of eIF4E present in an inactive complex with 4E-BP1 was increased 5-fold compared to freely fed control animals. One hour after refeeding a complete diet, the amount of 4E-BP1 bound to eIF4E was reduced to freely fed control values. Reduced association of the two proteins was the result of increased phosphorylation of 4E BP1. Refeeding a complete diet also stimulated the binding of eIF4E to eIF4G to form the active eIF4F complex. In liver, the amount of eIF4E associated with eIF4G, but not the amount of eIF4E associated with 4E-BP1, was altered by fasting and refeeding. Furthermore, in liver, but not in skeletal muscle, fasting and refeeding resulted in modulation of the phosphorylation state of eIF4E. Overall, the results suggest that protein synthesis may be differentially regulated in muscle and liver in response to fasting and refeeding. In muscle, protein synthesis is regulated through modulation of the binding of eIF4E to eIF4G and in liver through modulation of both phosphorylation of eIF4E as well as binding of eIF4E to eIF4G. PMID- 9398654 TI - Retinoic acid increases sodium/iodide symporter mRNA levels in human thyroid cancer cell lines and suppresses expression of functional symporter in nontransformed FRTL-5 rat thyroid cells. AB - Decreased iodide uptake in de-differentiated thyroid carcinomas impedes radioiodide therapy. RTPCR analysis revealed reduced expression of Na+/I- symporter (NIS) mRNA in human thyroid carcinomas as compared to normal thyroid. However, in follicular thyroid carcinoma cell lines FTC-133 and FTC-238, treatment with 1 microM all-trans retinoic acid (RA) markedly increased NIS mRNA levels. Anaplastic thyroid carcinoma cell lines HTh74 and C643 showed basal expression of NIS mRNA, but no RA-stimulation. All four cell lines contained the approximately 80 kD NIS protein as judged by Western blot, although they did not accumulate iodide. In contrast, in nontransformed rat FRTL-5 cells, 1 microM RA downregulated NIS mRNA levels, inhibited the TSH- or forskolin-triggered induction of NIS message after TSH-depletion, and reduced iodide uptake to 38% after 5 d. This divergent RA-responsivity of NIS may provide the means to target radioiodide to thyroid carcinomas by upregulating iodide transport into tumor tissue while simultaneously inhibiting iodide accumulation in normal thyrocytes and may thus re-establish the potential for radioiodide therapy. PMID- 9398655 TI - Stability of protease Q against autolysis and in sodium dodecyl sulfate and urea solutions. AB - Protease Q, a recently discovered subtilisin-like protease, exhibited unusual stability in 10% sodium dodecyl sulfate (SDS) and 8 M urea solutions. In 2% SDS it exhibited higher activity than the control, and in 10% SDS it retained 50% of its original activity when azocoll was used as substrate. Kinetic studies showed that the decrease in the activity of protease Q in SDS solutions followed a first order kinetics with a half-life of 446, 278, 78, and 24 h in 1%, 2%, 5%, and 10% SDS, respectively, at 25 degrees C. Protease Q was also stable against autolysis. In 20 mM Tris-HCl buffer (pH 8.3) containing 1.0 mM CaCl2, protease Q had a half life of 2822 h and 725 h at room temperature and at 37 degrees C, respectively. The enzyme was able to completely hydrolyze beta-lactoglobulin, beta-casein, and keratin in 8-10 M urea. PMID- 9398656 TI - Functional characterization of RNase H1 from Drosophila melanogaster. AB - We have cloned and functionally characterized the RNase H1 gene from D. melanogaster. The longest open reading frame consists of 5 exons that encode a 333 amino acid protein with a molecular mass of 37.1 kDa. This is the first demonstration of specific nuclease activity of a cloned RNase gene from a multicellular higher eukaryote. No additional proteins or cofactors are required for this nuclease activity. Comparison of Drosophila RNase H1 amino acid sequence to that of other cellular eukaryotic homologs reveals the presence of three evolutionarily distinct domains. The N- and C-terminal conserved domains are connected by a highly variable domain. The C-terminal domain has high amino acid similarity to bacterial RNase HI and the RNase H domain of retroviral reverse transcriptase, while the N-terminus, of unknown function, is similar to the P6 translational activator of caulimoviruses. PMID- 9398657 TI - Potent inactivator of alpha-chymotrypsin: 2,2-dimethyl-3-(N-4-cyanobenzoyl)amino 5-phenyl pentanoic anhydride. AB - We synthesized a novel potent alpha-chymotrypsin inactivator, 2,2-dimethyl-3-(N-4 cyanobenzoyl) amino-5-phenyl pentanoic anhydride, which fulfilled the criteria of a mechanism-based inactivator: first-order kinetics, irreversibility, saturation kinetics and substrate protection. The inactivation rate constant (kinact) and the enzyme-inhibitor dissociation constant (KI) were calculated to be 0.017s-1 and 0.071 microM, respectively (kinact/KI = 242,000 M-1s-1). These kinetic parameters indicate that this compound is one of the most powerful alpha chymotrypsin inactivators ever reported. The average number of alpha-chymotrypsin turnovers per inactivation (partition ratio) was calculated to be 1, which indicates that it is a stoichiometrically ideal inactivator of alpha chymotrypsin. We compared the IC50 values of this compound with those of several chymotrypsin-like serine proteinases (bovine alpha-chymotrypsin, recombinant human chymase and human neutrophil cathepsin G) and a metallo proteinase, rabbit angiotensin converting enzyme (ACE). Our compound, 2,2-dimethyl-3-(N-4 cyanobenzoyl) amino-5-phenyl pentanoic anhydride, inhibited bovine alpha chymotrypsin potently (IC50 = 1.0 (+/- 0.2) x 10(-9) M) as well as other chymotrypsin-like serine proteinase; recombinant human chymase (IC50 = 7.0 (+/- 1.0) x 10(-8) M) and human neutrophil cathepsin G (IC50 = 1.8 (+/- 0.2) x 10(-7) M). However, rabbit ACE was not inhibited by this compound (IC50 > 1 x 10(-4) M). PMID- 9398658 TI - Lysophosphatidic acid inhibits epidermal-growth-factor-induced Stat1 signaling in human epidermoid carcinoma A431 cells. AB - Lysophosphatidic acid (LPA) is a lipid mediator which acts on its putative G protein-coupled receptor (GPCR). Recently, activation of signal transducers and activators of transcription (STATs) mediated by GPCR has been reported. In this study, we examined the effect of LPA on STAT activation using the electrophoretic mobility shift assays and the heterologous promoter analysis in human epidermoid carcinoma A431 cells. We found that LPA inhibited epidermal growth factor (EGF) induced Stat1 activation in a concentration-dependent manner. Other phospholipase C (PLC)-coupled GPCR agonists, bradykinin and ATP, also inhibited Stat1 activation. This inhibitory effect of LPA was completely mimicked by an activator of protein kinase C (PKC), a PLC-downstream effector. These findings suggest that the inhibitory effect on EGF-induced Stat1 activation may be a general characteristic of PLC-coupled GPCRs and PKC pathway may be mainly associated with this inhibitory effect. This is the first evidence showing that GPCR agonists inhibit the Janus kinase-independent Stat1 activation induced by receptor tyrosine kinase. PMID- 9398659 TI - Interferon induces up-regulation of Spi-1/PU.1 in human leukemia K562 cells. AB - The human K562 cell line is derived from a chronic myelogenous leukemia in blastic crisis. Treatment of K562 cells with interferons alpha, beta or gamma resulted in inhibition of cell proliferation. Spi-1/PU.1 is a transcription factor of the Ets family which is required for normal hematopoyesis. We have found that spi-1 mRNA and protein as well as Spi-1-DNA binding activity increase after exposure of K562 cells to interferons. The increase in spi-1 expression ranged from 4- to 8-fold with the different interferons. K562 cells can be differentiated in vitro towards erythroid cells or monocyte-macrophage cells. Interestingly, the regulation of spi-1 by interferon-alpha depended on the differentiated phenotype of K562 cells: interferon-alpha failed to induce spi-1 in erythroid differentiated cells, whereas it induced spi-1 in monocyte macrophage differentiated cells. The results suggest a role for Spi-1 in the cytostatic response to interferons. PMID- 9398660 TI - Distance between the basic group of the amino acid residue's side chain in position P1 of trypsin inhibitor CMTI-III and Asp189 in the substrate pocket of trypsin has an essential influence on the inhibitory activity. AB - Three new analogues of trypsin inhibitor CMTI-III were synthesized by the solid phase method: [Lys5]-CMTI-III, [Orn5]CMTI-III and [Dab5]CMTI-III. Only one analogue with L-lysine residue in position P1 showed inhibitory activity of the same order of magnitude as did wild CMTI-III. Two remaining analogues were completely inactive. A conclusion was drawn that the distance between the basic group of the amino acid residue's side chain in position P1 of the trypsin inhibitor CMTI-III and Asp189 in the substrate pocket of trypsin plays an essential role for the trypsin-inhibitor interaction. PMID- 9398661 TI - Organization of G proteins and adenylyl cyclase at the plasma membrane. AB - There is mounting evidence for the organization and compartmentation of signaling molecules at the plasma membrane. We find that hormone-sensitive adenylyl cyclase activity is enriched in a subset of regulatory G protein-containing fractions of the plasma membrane. These subfractions resemble, in low buoyant density, structures of the plasma membrane termed caveolae. Immunofluorescence experiments revealed a punctate pattern of G protein alpha and beta subunits, consistent with concentration of these proteins at distinct sites on the plasma membrane. Partial coincidence of localization of G protein alpha subunits with caveolin (a marker for caveolae) was observed by double immunofluorescence. Results of immunogold electron microscopy suggest that some G protein is associated with invaginated caveolae, but most of the protein resides in irregular structures of the plasma membrane that could not be identified morphologically. Because regulated adenylyl cyclase activity is present in low-density subfractions of plasma membrane from a cell type (S49 lymphoma) that does not express caveolin, this protein is not required for organization of the adenylyl cyclase system. The data suggest that hormone-sensitive adenylyl cyclase systems are localized in a specialized subdomain of the plasma membrane that may optimize the efficiency and fidelity of signal transduction. PMID- 9398662 TI - RanGTP targets p97 to RanBP2, a filamentous protein localized at the cytoplasmic periphery of the nuclear pore complex. AB - RanBP2, a protein containing FG repeat motifs and four binding sites for the guanosine triphosphatase Ran, is localized at the cytoplasmic periphery of the nuclear pore complex (NPC) and is believed to play a critical role in nuclear protein import. We purified RanBP2 from rat liver nuclear envelopes and examined its structural and biochemical properties. Electron microscopy showed that RanBP2 forms a flexible filamentous molecule with a length of approximately 36 nm, suggesting that it comprises a major portion of the cytoplasmic fibrils implicated in initial binding of import substrates to the NPC. Using in vitro assays, we characterized the ability of RanBP2 to bind p97, a cytosolic factor implicated in the association of the nuclear localization signal receptor with the NPC. We found that RanGTP promotes the binding of p97 to RanBP2, whereas it inhibits the binding of p97 to other FG repeat nucleoporins. These data suggest that RanGTP acts to specifically target p97 to RanBP2, where p97 may support the binding of an nuclear localization signal receptor/substrate complex to RanBP2 in an early step of nuclear import. PMID- 9398663 TI - Gradual phenotypic conversion associated with immortalization of cultured human mammary epithelial cells. AB - Examination of the process of immortal transformation in early passages of two human mammary epithelial cell (HMEC) lines suggests the involvement of an epigenetic step. These lines, 184A1 and 184B5, arose after in vitro exposure of finite lifespan 184 HMEC to a chemical carcinogen, and both are clonally derived. Although early-passage mass cultures of 184A1 and 184B5 maintained continuous slow growth, most individual cells lost proliferative ability. Uniform good growth did not occur until 20-30 passages after the lines first appeared. Early passage cultures expressed little or no telomerase activity and telomeres continued to shorten with increasing passage. Telomerase activity was first detected when the telomeres became critically short, and activity levels gradually increased thereafter. Early-passage cultures had little or no ability to maintain growth in transforming growth factor-beta (TGFbeta); however, both mass cultures and clonal isolates showed a very gradual increase in the number of cells displaying progressively increased ability to maintain growth in TGFbeta. A strong correlation between capacity to maintain growth in the presence of TGFbeta and expression of telomerase activity was observed. We have used the term "conversion" to describe this process of gradual acquisition of increased growth capacity in the absence or presence of TGFbeta and reactivation of telomerase. We speculate that the development of extremely short telomeres may result in gradual, epigenetic-based changes in gene expression. Understanding the underlying mechanisms of HMEC conversion in vitro may provide new insight into the process of carcinogenic progression in vivo and offer novel modes for therapeutic intervention. PMID- 9398664 TI - Dissociation of Oct-1 from the nuclear peripheral structure induces the cellular aging-associated collagenase gene expression. AB - The cellular aging-associated transcriptional repressor that we previously named as Orpheus was identical to Oct-1, a member of the POU domain family. Oct-1 represses the collagenase gene, one of the cellular aging-associated genes, by interacting with an AT-rich cis-element in the upstream of the gene in preimmortalized cells at earlier population-doubling levels and in immortalized cells. In these stages of cells, considerable fractions of the Oct-1 protein were prominently localized in the nuclear periphery and colocalized with lamin B. During the cellular aging process, however, this subspecies of Oct-1 disappeared from the nuclear periphery. The cells lacking the nuclear peripheral Oct-1 protein exhibited strong collagenase expression and carried typical senescent morphologies. Concomitantly, the binding activity and the amount of nuclear Oct-1 protein were reduced in the aging process and resumed after immortalization. However, the whole cellular amounts of Oct-1 protein were not significantly changed during either process. Thus, the cellular aging-associated genes including the collagenase gene seemed to be derepressed by the dissociation of Oct-1 protein from the nuclear peripheral structure. Oct-1 may form a transcriptional repressive apparatus by anchoring nuclear matrix attachment regions onto the nuclear lamina in the nuclear periphery. PMID- 9398665 TI - SET1, a yeast member of the trithorax family, functions in transcriptional silencing and diverse cellular processes. AB - The trithorax gene family contains members implicated in the control of transcription, development, chromosome structure, and human leukemia. A feature shared by some family members, and by other proteins that function in chromatin mediated transcriptional regulation, is the presence of a 130- to 140-amino acid motif dubbed the SET or Tromo domain. Here we present analysis of SET1, a yeast member of the trithorax gene family that was identified by sequence inspection to encode a 1080-amino acid protein with a C-terminal SET domain. In addition to its SET domain, which is 40-50% identical to those previously characterized, SET1 also shares dispersed but significant similarity to Drosophila and human trithorax homologues. To understand SET1 function(s), we created a null mutant. Mutant strains, although viable, are defective in transcriptional silencing of the silent mating-type loci and telomeres. The telomeric silencing defect is rescued not only by full-length episomal SET1 but also by the conserved SET domain of SET1. set1 mutant strains display other phenotypes including morphological abnormalities, stationary phase defects, and growth and sporulation defects. Candidate genes that may interact with SET1 include those with functions in transcription, growth, and cell cycle control. These data suggest that yeast SET1, like its SET domain counterparts in other organisms, functions in diverse biological processes including transcription and chromatin structure. PMID- 9398667 TI - alphavbeta3 integrin mediates the cell-adhesive capacity and biological activity of basic fibroblast growth factor (FGF-2) in cultured endothelial cells. AB - Fibroblast growth factor-2 (FGF-2) immobilized on non-tissue culture plastic promotes adhesion and spreading of bovine and human endothelial cells that are inhibited by anti-FGF-2 antibody. Heat-inactivated FGF-2 retains its cell adhesive activity despite its incapacity to bind to tyrosine-kinase FGF receptors or to cell-surface heparan sulfate proteoglycans. Recombinant glutathione-S transferase-FGF-2 chimeras and synthetic FGF-2 fragments identify two cell adhesive domains in FGF-2 corresponding to amino acid sequences 38-61 and 82-101. Both regions are distinct from the FGF-receptor-binding domain of FGF-2 and contain a DGR sequence that is the inverse of the RGD cell-recognition sequence. Calcium deprivation, RGD-containing eptapeptides, soluble vitronectin (VN), but not fibronectin (FN), inhibit cell adhesion to FGF-2. Conversely, soluble FGF-2 prevents cell adhesion to VN but not FN, thus implicating VN receptor in the cell adhesive activity of FGF-2. Accordingly, monoclonal and polyclonal anti alphavbeta3 antibodies prevent cell adhesion to FGF-2. Also, purified human alphavbeta3 binds to immobilized FGF-2 in a cation-dependent manner, and this interaction is competed by soluble VN but not by soluble FN. Finally, anti alphavbeta3 monoclonal and polyclonal antibodies specifically inhibit mitogenesis and urokinase-type plasminogen activator (uPA) up-regulation induced by free FGF 2 in endothelial cells adherent to tissue culture plastic. These data demonstrate that FGF-2 interacts with alphavbeta3 integrin and that this interaction mediates the capacity of the angiogenic growth factor to induce cell adhesion, mitogenesis, and uPA up-regulation in endothelial cells. PMID- 9398666 TI - Inhibition of RhoA translocation and calcium sensitization by in vivo ADP ribosylation with the chimeric toxin DC3B. AB - Pretreatment of intact rabbit portal vein smooth muscle with the chimeric toxin DC3B (10(-6) M, 48 h; ; ) ADP-ribosylated endogenous RhoA, including cytosolic RhoA complexed with rhoGDI, and inhibited the tonic phase of phenylephrine induced contraction and the Ca2+-sensitization of force by phenylephrine, endothelin and guanosine triphosphate (GTP)gammaS, but did not inhibit Ca2+ sensitization by phorbol dibutyrate. DC3B also inhibited GTPgammaS-induced translocation of cytosolic RhoA () to the membrane fraction. In DC3B-treated muscles the small fraction of membrane-associated RhoA could be immunoprecipitated, even after exposure to GTPgammaS, which prevents immunoprecipitation of non-ADP-ribosylated RhoA. Dissociation of cytosolic RhoA rhoGDI complexes with SDS restored the immunoprecipitability and ADP ribosylatability of RhoA, indicating that both the ADP-ribosylation site (Asn 41) and RhoA insert loop (Wei et al., 1997) are masked by rhoGDI and that the long axes of the two proteins are in parallel in the heterodimer. We conclude that RhoA plays a significant role in G-protein-, but not protein kinase C-mediated, Ca2+ sensitization and that ADP ribosylation inhibits in vivo the Ca2+ sensitizing effect of RhoA by interfering with its binding to a membrane associated effector. PMID- 9398668 TI - Interaction of class I human leukocyte antigen (HLA-I) molecules with insulin receptors and its effect on the insulin-signaling cascade. AB - Insulin receptor (IR) and class I major histocompatibility complex molecules associate with one another in cell membranes, but the functional consequences of this association are not defined. We found that IR and human class I molecules (HLA-I) associate in liposome membranes and that the affinity of IR for insulin and its tyrosine kinase activity increase as the HLA:IR ratio increases over the range 1:1 to 20:1. The same relationship between HLA:IR and IR function was found in a series of B-LCL cell lines. The association of HLA-I and IR depends upon the presence of free HLA heavy chains. All of the effects noted were reduced or abrogated if liposomes or cells were incubated with excess HLA-I light chain, beta2-microglobulin. Increasing HLA:IR also enhanced phosphorylation of insulin receptor substrate-1 and the activation of phosphoinositide 3-kinase. HLA-I molecules themselves were phosphorylated on tyrosine and associated with phosphoinositide 3-kinase when B-LCL were stimulated with insulin. PMID- 9398669 TI - A WD repeat protein controls the cell cycle and differentiation by negatively regulating Cdc2/B-type cyclin complexes. AB - In the fission yeast Schizosaccharomyces pombe, p34(cdc2) plays a central role controlling the cell cycle. We recently isolated a new gene named srw1(+), capable of encoding a WD repeat protein, as a multicopy suppressor of hyperactivated p34(cdc2). Cells lacking srw1(+) are sterile and defective in cell cycle controls. When starved for nitrogen source, they fail to effectively arrest in G1 and die of accelerated mitotic catastrophe if regulation of p34(cdc2)/Cdc13 by inhibitory tyrosine phosphorylation is compromised by partial inactivation of Wee1 kinase. Fertility is restored to the disruptant by deletion of Cig2 B-type cyclin or slight inactivation of p34(cdc2). srw1(+) shares functional similarity with rum1(+), having abilities to induce endoreplication and restore fertility to rum1 disruptants. In the srw1 disruptant, Cdc13 fails to be degraded when cells are starved for nitrogen. We conclude that Srw1 controls differentiation and cell cycling at least by negatively regulating Cig2- and Cdc13-associated p34(cdc2) and that one of its roles is to down-regulate the level of the mitotic cyclin particularly in nitrogen-poor environments. PMID- 9398670 TI - Yeast cycloheximide-resistant crl mutants are proteasome mutants defective in protein degradation. AB - In 1988 McCusker and Haber generated a series of mutants which are resistant to the minimum inhibitory concentration of the protein synthesis inhibitor cycloheximide. These cycloheximide-resistant, temperature-sensitive (crl) mutants, in addition, exhibited other pleiotropic phenotypes, e.g., incorrect response to starvation, hypersensitivity against amino acid analogues, and other protein synthesis inhibitors. Temperature sensitivity of one of these mutants, crl3-2, had been found to be suppressed by a mutation, SCL1-1, which resided in an alpha-type subunit of the 20S proteasome. We cloned the CRL3 gene by complementation and found CRL3 to be identical to the SUG1/CIM3 gene coding for a subunit of the 19S cap complex of the 26S proteasome. Another mutation, crl21, revealed to be allelic with the 20S proteasomal gene PRE3. crl3-2 and crl21 mutant cells show significant defects in proteasome-dependent proteolysis, whereas the SCL1-1 suppressor mutation causes partial restoration of crl3-2 induced proteolytic defects. Notably, cycloheximide resistance was also detected for other proteolytically deficient proteasome mutants (pre1-1, pre2-1, pre3-1, pre4-1). Moreover, proteasomal genes were found within genomic sequences of 9 of 13 chromosomal loci to which crl mutations had been mapped. We therefore assume that most if not all crl mutations reside in the proteasome and that phenotypes found are a result of defective protein degradation. PMID- 9398672 TI - Gene conversion of major histocompatibility complex genes in the mouse spermatogenesis is a premeiotic event. AB - The molecular genetic mechanism of gene conversion in higher eukaryotes remains unknown. We find it of considerable interest to determine when during spermatogenesis gene conversion occurs. We have therefore purified pachytene spermatocytes and haploid spermatocytes from adult mice and analyzed these fractions for the presence of gene conversion products resulting from the transfer between the major histocompatibility complex class II genes Ebd and Abk in a polymerase chain reaction assay. We have further isolated spermatogenic cells from prepubescent mice and analyzed them for the presence of the same gene conversion products. We can detect gene conversion products in testis cells as early as in 8-d-old mice where the only existing spermatogenic cells are spermatogonia. The frequency of gene conversion products remains the same as the cells reach meiosis in 18-d-old mice, and is unchanged after meiosis is completed in haploid spermatocytes. Gene conversion of this specific fragment therefore appears to be a premeiotic event and, consequently, relies on genetic mechanisms other than normal meiotic recombination. PMID- 9398671 TI - Differential in vivo regulation of steroid hormone receptor activation by Cdc37p. AB - The CDC37 gene is essential for the activity of p60(v-src) when expressed in yeast cells. Since the activation pathway for p60(v-src) and steroid hormone receptors is similar, the present study analyzed the hormone-dependent transactivation by androgen receptors and glucocorticoid receptors in yeast cells expressing a mutant version of the CDC37 gene. In this mutant, hormone-dependent transactivation by androgen receptors was defective at both permissive and restrictive temperatures, although transactivation by glucocorticoid receptors was mildly defective only at the restrictive temperature. Cdc37p appears to function via the androgen receptor ligand-binding domain, although it does not influence receptor hormone-binding affinity. Models for Cdc37p regulation of steroid hormone receptors are discussed. PMID- 9398673 TI - Characterization of Saccharomyces cerevisiae dna2 mutants suggests a role for the helicase late in S phase. AB - The TOR proteins, originally identified as targets of the immunosuppressant rapamycin, contain an ATM-like "lipid kinase" domain and are required for early G1 progression in eukaryotes. Using a screen to identify Saccharomyces cerevisiae mutants requiring overexpression of Tor1p for viability, we have isolated mutations in a gene we call ROT1 (requires overexpression of Tor1p). This gene is identical to DNA2, encoding a helicase required for DNA replication. As with its role in cell cycle progression, both the N-terminal and C-terminal regions, as well as the kinase domain of Tor1p, are required for rescue of dna2 mutants. Dna2 mutants are also rescued by Tor2p and show synthetic lethality with tor1 deletion mutants under specific conditions. Temperature-sensitive (Ts) dna2 mutants arrest irreversibly at G2/M in a RAD9- and MEC1-dependent manner, suggesting that Dna2p has a role in S phase. Frequencies of mitotic recombination and chromosome loss are elevated in dna2 mutants, also supporting a role for the protein in DNA synthesis. Temperature-shift experiments indicate that Dna2p functions during late S phase, although dna2 mutants are not deficient in bulk DNA synthesis. These data suggest that Dna2p is not required for replication fork progression but may be needed for a later event such as Okazaki fragment maturation. PMID- 9398675 TI - Ubiquitously expressed dynamin-II has a higher intrinsic GTPase activity and a greater propensity for self-assembly than neuronal dynamin-I. AB - To begin to understand mechanistic differences in endocytosis in neurons and nonneuronal cells, we have compared the biochemical properties of the ubiquitously expressed dynamin-II isoform with those of neuron-specific dynamin I. Like dynamin-I, dynamin-II is specifically localized to and highly concentrated in coated pits on the plasma membrane and can assemble in vitro into rings and helical arrays. As expected, the two closely related isoforms share a similar mechanism for GTP hydrolysis: both are stimulated in vitro by self assembly and by interaction with microtubules or the SH3 domain-containing protein, grb2. Deletion of the C-terminal proline/arginine-rich domain from either isoform abrogates self-assembly and assembly-dependent increases in GTP hydrolysis. However, dynamin-II exhibits a approximately threefold higher rate of intrinsic GTP hydrolysis and higher affinity for GTP than dynamin-I. Strikingly, the stimulated GTPase activity of dynamin-II can be >40-fold higher than dynamin I, due principally to its greater propensity for self-assembly and the increased resistance of assembled dynamin-II to GTP-triggered disassembly. These results are consistent with the hypothesis that self-assembly is a major regulator of dynamin GTPase activity and that the intrinsic rate of GTP hydrolysis reflects a dynamic, GTP-dependent equilibrium of assembly and disassembly. PMID- 9398676 TI - Modulatory roles for integrin activation and the synergy site of fibronectin during matrix assembly. AB - Initiation of fibronectin (FN) matrix assembly is dependent on specific interactions between FN and cell surface integrin receptors. Here, we show that de novo FN matrix assembly exhibits a slow phase during initiation of fibrillogenesis followed by a more rapid growth phase. Mn2+, which acts by enhancing integrin function, increased the rate of FN fibril growth, but only after the initial lag phase. The RGD cell-binding sequence in type III repeat 10 is an absolute requirement for initiation by alpha5beta1 integrin. To investigate the role of the cell-binding synergy site in the adjacent repeat III9, a full length recombinant FN containing a synergy mutation, FN(syn-), was tested for its ability to form fibrils. Mutation of this site drastically reduced FN assembly by CHOalpha5 cells. Only sparse short fibrils were formed even after prolonged incubation, indicating that FN(syn-) is defective in progression of the assembly process. These results show that the synergy site is essential for alpha5beta1 mediated accumulation of a FN matrix. However, the incorporation of FN(syn-) into fibrils and the deoxycholate-insoluble matrix could be stimulated by Mn2+. Therefore, exogenous activation of integrin receptors can overcome the requirement for FN's synergy site as well as modulate the rate of FN matrix formation. PMID- 9398674 TI - Derepressed hyphal growth and reduced virulence in a VH1 family-related protein phosphatase mutant of the human pathogen Candida albicans. AB - Mitogen-activated protein (MAP) kinases are pivotal components of eukaryotic signaling cascades. Phosphorylation of tyrosine and threonine residues activates MAP kinases, but either dual-specificity or monospecificity phosphatases can inactivate them. The Candida albicans CPP1 gene, a structural member of the VH1 family of dual- specificity phosphatases, was previously cloned by its ability to block the pheromone response MAP kinase cascade in Saccharomyces cerevisiae. Cpp1p inactivated mammalian MAP kinases in vitro and acted as a tyrosine-specific enzyme. In C. albicans a MAP kinase cascade can trigger the transition from the budding yeast form to a more invasive filamentous form. Disruption of the CPP1 gene in C. albicans derepressed the yeast to hyphal transition at ambient temperatures, on solid surfaces. A hyphal growth rate defect under physiological conditions in vitro was also observed and could explain a reduction in virulence associated with reduced fungal burden in the kidneys seen in a systemic mouse model. A hyper-hyphal pathway may thus have some detrimental effects on C. albicans cells. Disruption of the MAP kinase homologue CEK1 suppressed the morphological effects of the CPP1 disruption in C. albicans. The results presented here demonstrate the biological importance of a tyrosine phosphatase in cell-fate decisions and virulence in C. albicans. PMID- 9398677 TI - A mutational analysis identifies three functional regions of the spindle pole component Spc110p in Saccharomyces cerevisiae. AB - The central coiled coil of the essential spindle pole component Spc110p spans the distance between the central and inner plaques of the Saccharomyces cerevisiae spindle pole body (SPB). The carboxy terminus of Spc110p, which binds calmodulin, resides at the central plaque, and the amino terminus resides at the inner plaque from which nuclear microtubules originate. To dissect the functions of Spc110p, we created temperature-sensitive mutations in the amino and carboxy termini. Analysis of the temperature-sensitive spc110 mutations and intragenic complementation analysis of the spc110 alleles defined three functional regions of Spc110p. Region I is located at the amino terminus. Region II is located at the carboxy-terminal end of the coiled coil, and region III is the previously defined calmodulin-binding site. Overexpression of SPC98 suppresses the temperature sensitivity conferred by mutations in region I but not the phenotypes conferred by mutations in the other two regions, suggesting that the amino terminus of Spc110p is involved in an interaction with the gamma-tubulin complex composed of Spc97p, Spc98p, and Tub4p. Mutations in region II lead to loss of SPB integrity during mitosis, suggesting that this region is required for the stable attachment of Spc110p to the central plaque. Our results strongly argue that Spc110p links the gamma-tubulin complex to the central plaque of the SPB. PMID- 9398678 TI - A T42A Ran mutation: differential interactions with effectors and regulators, and defect in nuclear protein import. AB - Ran, the small, predominantly nuclear GTPase, has been implicated in the regulation of a variety of cellular processes including cell cycle progression, nuclear-cytoplasmic trafficking of RNA and protein, nuclear structure, and DNA synthesis. It is not known whether Ran functions directly in each process or whether many of its roles may be secondary to a direct role in only one, for example, nuclear protein import. To identify biochemical links between Ran and its functional target(s), we have generated and examined the properties of a putative Ran effector mutation, T42A-Ran. T42A-Ran binds guanine nucleotides as well as wild-type Ran and responds as well as wild-type Ran to GTP or GDP exchange stimulated by the Ran-specific guanine nucleotide exchange factor, RCC1. T42A-Ran.GDP also retains the ability to bind p10/NTF2, a component of the nuclear import pathway. In contrast to wild-type Ran, T42A-Ran.GTP binds very weakly or not detectably to three proposed Ran effectors, Ran-binding protein 1 (RanBP1), Ran-binding protein 2 (RanBP2, a nucleoporin), and karyopherin beta (a component of the nuclear protein import pathway), and is not stimulated to hydrolyze bound GTP by Ran GTPase-activating protein, RanGAP1. Also in contrast to wild-type Ran, T42A-Ran does not stimulate nuclear protein import in a digitonin permeabilized cell assay and also inhibits wild-type Ran function in this system. However, the T42A mutation does not block the docking of karyophilic substrates at the nuclear pore. These properties of T42A-Ran are consistent with its classification as an effector mutant and define the exposed region of Ran containing the mutation as a probable effector loop. PMID- 9398679 TI - Myosin heavy chain phosphorylation sites regulate myosin localization during cytokinesis in live cells. AB - Conventional myosin II plays a fundamental role in the process of cytokinesis where, in the form of bipolar thick filaments, it is thought to be the molecular motor that generates the force necessary to divide the cell. In Dictyostelium, the formation of thick filaments is regulated by the phosphorylation of three threonine residues in the tail region of the myosin heavy chain. We report here on the effects of this regulation on the localization of myosin in live cells undergoing cytokinesis. We imaged fusion proteins of the green-fluorescent protein with wild-type myosin and with myosins where the three critical threonines had been changed to either alanine or aspartic acid. We provide evidence that thick filament formation is required for the accumulation of myosin in the cleavage furrow and that if thick filaments are overproduced, this accumulation is markedly enhanced. This suggests that myosin localization in dividing cells is regulated by myosin heavy chain phosphorylation. PMID- 9398680 TI - On the role of myosin-II in cytokinesis: division of Dictyostelium cells under adhesive and nonadhesive conditions. AB - We have investigated the role of myosin in cytokinesis in Dictyostelium cells by examining cells under both adhesive and nonadhesive conditions. On an adhesive surface, both wild-type and myosin-null cells undergo the normal processes of mitotic rounding, cell elongation, polar ruffling, furrow ingression, and separation of daughter cells. When cells are denied adhesion through culturing in suspension or on a hydrophobic surface, wild-type cells undergo these same processes. However, cells lacking myosin round up and polar ruffle, but fail to elongate, furrow, or divide. These differences show that cell division can be driven by two mechanisms that we term Cytokinesis A, which requires myosin, and Cytokinesis B, which is cell adhesion dependent. We have used these approaches to examine cells expressing a myosin whose two light chain-binding sites were deleted (DeltaBLCBS-myosin). Although this myosin is a slower motor than wild type myosin and has constitutively high activity due to the abolition of regulation by light-chain phosphorylation, cells expressing DeltaBLCBS-myosin were previously shown to divide in suspension (Uyeda et al., 1996). However, we suspected their behavior during cytokinesis to be different from wild-type cells given the large alteration in their myosin. Surprisingly, DeltaBLCBS-myosin undergoes relatively normal spatial and temporal changes in localization during mitosis. Furthermore, the rate of furrow progression in cells expressing a DeltaBLCBS-myosin is similar to that in wild-type cells. PMID- 9398681 TI - Accumulation of major histocompatibility complex class II molecules in mast cell secretory granules and their release upon degranulation. AB - To investigate the relationship between major histocompatibility complex (MHC) class II compartments, secretory granules, and secretory lysosomes, we analyzed the localization and fate of MHC class II molecules in mast cells. In bone marrow derived mast cells, the bulk of MHC class II molecules is contained in two distinct compartments, with features of both lysosomal compartments and secretory granules defined by their protein content and their accessibility to endocytic tracers. Type I granules display internal membrane vesicles and are accessed by exogenous molecules after a time lag of 20 min; type II granules are reached by the endocytic tracer later and possess a serotonin-rich electron-dense core surrounded by a multivesicular domain. In these type I and type II granules, MHC class II molecules, mannose-6-phosphate receptors and lysosomal membrane proteins (lamp1 and lamp2) localize to small intralumenal vesicles. These 60-80-nm vesicles are released along with inflammatory mediators during mast cell degranulation triggered by IgE-antigen complexes. These observations emphasize the intimate connection between the endocytic and secretory pathways in cells of the hematopoietic lineage which allows regulated secretion of the contents of secretory lysosomes, including membrane proteins associated with small vesicles. PMID- 9398682 TI - Defining extracellular integrin alpha-chain sites that affect cell adhesion and adhesion strengthening without altering soluble ligand binding. AB - It was previously shown that mutations of integrin alpha4 chain sites, within putative EF-hand-type divalent cation-binding domains, each caused a marked reduction in alpha4beta1-dependent cell adhesion. Some reports have suggested that alpha-chain "EF-hand" sites may interact directly with ligands. However, we show here that mutations of three different alpha4 "EF-hand" sites each had no effect on binding of soluble monovalent or bivalent vascular cell adhesion molecule 1 whether measured indirectly or directly. Furthermore, these mutations had minimal effect on alpha4beta1-dependent cell tethering to vascular cell adhesion molecule 1 under shear. However, EF-hand mutants did show severe impairments in cellular resistance to detachment under shear flow. Thus, mutation of integrin alpha4 "EF-hand-like" sites may impair 1) static cell adhesion and 2) adhesion strengthening under shear flow by a mechanism that does not involve alterations of initial ligand binding. PMID- 9398683 TI - Characterization of a novel yeast SNARE protein implicated in Golgi retrograde traffic. AB - The protein trafficking machinery of eukaryotic cells is employed for protein secretion and for the localization of resident proteins of the exocytic and endocytic pathways. Protein transit between organelles is mediated by transport vesicles that bear integral membrane proteins (v-SNAREs) which selectively interact with similar proteins on the target membrane (t-SNAREs), resulting in a docked vesicle. A novel Saccharomyces cerevisiae SNARE protein, which has been termed Vti1p, was identified by its sequence similarity to known SNAREs. Vti1p is a predominantly Golgi-localized 25-kDa type II integral membrane protein that is essential for yeast viability. Vti1p can bind Sec17p (yeast SNAP) and enter into a Sec18p (NSF)-sensitive complex with the cis-Golgi t-SNARE Sed5p. This Sed5p/Vti1p complex is distinct from the previously described Sed5p/Sec22p anterograde vesicle docking complex. Depletion of Vti1p in vivo causes a defect in the transport of the vacuolar protein carboxypeptidase Y through the Golgi. Temperature-sensitive mutants of Vti1p show a similar carboxypeptidase Y trafficking defect, but the secretion of invertase and gp400/hsp150 is not significantly affected. The temperature-sensitive vti1 growth defect can be rescued by the overexpression of the v-SNARE, Ykt6p, which physically interacts with Vti1p. We propose that Vti1p, along with Ykt6p and perhaps Sft1p, acts as a retrograde v-SNARE capable of interacting with the cis-Golgi t-SNARE Sed5p. PMID- 9398684 TI - BIM1 encodes a microtubule-binding protein in yeast. AB - A previously uncharacterized yeast gene (YER016w) that we have named BIM1 (binding to microtubules) was obtained from a two-hybrid screen of a yeast cDNA library using as bait the entire coding sequence of TUB1 (encoding alpha tubulin). Deletion of BIM1 results in a strong bilateral karyogamy defect, hypersensitivity to benomyl, and aberrant spindle behavior, all phenotypes associated with mutations affecting microtubules in yeast, and inviability at extreme temperatures (i.e., >/=37 degrees C or 40 nmol/L). There was no difference in peak post-absorptive serum cortisol or area under the concentration-time curve, and only three patients had a peak serum cortisol of more than 700 nmol/L. There was no difference in present height Z-score (-0.96; -0.24; -0.6), height Z-score at age 2 yr (-1.5; +0.4; -1.3), or current growth velocity Z-score (-0.1; +1.2; -2.2) between the groups, but bone maturation Z-score was significantly delayed (-1.63) in the tight control group and advanced (+0.8) in the poor control group. Present height was highly correlated (r = 0.8) with height at age 2 yr. Serum calcium, phosphorus, alkaline phosphatase, parathormone, and 25OH-vitamin D levels were all normal. There was no difference between the groups in age-corrected vertebral bone mineral density, and no difference in serum osteocalcin, procollagen peptide, or collagen C-terminal telopeptide, nor in urinary amino-terminal telopeptide. The data suggest that current methods of cortisol replacement do not significantly influence bone formation, resorption or density during childhood and therefore should not contribute to adult osteoporosis. The possibility remains that hypercortisolemia during infancy produces the short stature and delayed bone maturation that are present by the age 2 yr. PMID- 9398690 TI - Resistance to TSH in patients with normal TSH receptors--where do we turn when "Sutton's law" proves false? PMID- 9398691 TI - Resistance to thyrotropin (TSH) in three families is not associated with mutations in the TSH receptor or TSH. AB - Resistance to TSH (RTSH) is a recently described syndrome of reduced sensitivity to TSH that manifests as euthyroid hyperthyrotropinemia. It is usually identified at birth during routine neonatal screening for congenital hypothyroidism. In less than 2 yr, 13 subjects with RTSH belonging to 8 families have been reported, and all were shown to harbor mutations in the TSH receptor (TSHR) gene. We now report the occurrence of RTSH in 3 unrelated families. Contrary to previous reports, the inheritance of RTSH in 2 of the families was dominant rather than recessive and was not associated with abnormalities in the TSHR gene. Abnormalities in the TSHR gene were excluded by sequencing all coding sequences, exon/intron junctions, and the promoter region of the gene. Furthermore, the involvement of the TSHR in the manifestation of the RTSH phenotype was excluded in 2 families by linkage analysis using intragenic polymorphic markers. We excluded defects in the TSH beta-subunit by sequencing its gene and by showing that the circulating TSH in affected subjects from all families had normal bioactivity. Also, no abnormalities were found in the Gs alpha gene of one family analyzed by GC clamped denaturing gradient gel electrophoresis. This study shows that RTSH may be a manifestation of several different genetic defects that requires the exploration of other candidate genes involved in the TSH-TSHR-Gs alpha cascade and genes participating in its regulation. PMID- 9398692 TI - The binding protein's binding protein--clinical applications of acid-labile subunit (ALS) measurement. PMID- 9398693 TI - Acid-labile subunit of human insulin-like growth factor-binding protein complex: measurement, molecular, and clinical evaluation. AB - Although the acid-labile subunit (ALS) of the approximately 150-kDa insulin-like growth factor (IGF)-binding protein (IGFBP) complex was described over a decade ago, details of ALS physiology have remained largely uncertain. We evaluated antibodies to synthetic human ALS and constructed a noncompetitive ALS enzyme linked immunosorbent assay. Whereas uncomplexed ALS is directly measured, determination of total levels required sample pretreatment with SDS, which was found to optimally dissociate complexed ALS and significantly enhance ALS immunoreactivity. ALS in random adult sera was approximately 50% uncomplexed, and samples devoid of complexed ALS by immunoaffinity separation contained about 54% of the total levels. Serum ALS fractionated by gel filtration high performance liquid chromatography eluted in a single peak at approximately 150 kDa with IGF-I and IGFBP-3, but appeared at about 400-500 kDa after sample acidification and fractionation under acidic condition. The unexpected shift in ALS immunoreactivity remained unchanged when acid-neutralized or SDS-treated samples were fractionated under neutral pH and was reproducible when the 150-kDa complex was isolated, treated with acid or SDS, and rechromatographed. ALS in adult sera more tightly correlated with IGFBP-3 than IGF-I or IGF-II. The total levels (mean +/- SD) were 16.7 +/- 3.7 mg/L in 22 normal subjects, 28.3 +/- 8.1 mg/L in 20 acromegalic patients, and 9.5 +/- 3.8 in 32 GH-deficient adults. Little or no ALS was detectable in amniotic fluid, cerebrospinal fluid, seminal plasma, or milk, whereas high levels were present in synovial fluid. The development of ALS enzyme linked immunosorbent assay should greatly facilitate further investigations of this unique glycoprotein. PMID- 9398694 TI - Multiple beneficial effects of estrogen on lipoprotein metabolism. PMID- 9398695 TI - Effect of estrogen on very low density lipoprotein and low density lipoprotein subclass metabolism in postmenopausal women. AB - Estrogen decreases low density lipoprotein (LDL) particle size, and smaller LDL particles are associated with coronary atherosclerosis. To understand the metabolic basis for this change, we studied the effect of oral 17 beta-estradiol (2 mg/day) on apolipoprotein B-100 (apoB) metabolism, in eight healthy postmenopausal women. The study was a randomized, double blinded, placebo controlled, cross-over trial with intervention sequences of 6 weeks each. ApoB in very low density lipoprotein, intermediate density lipoprotein, and LDL subclasses was endogenously labeled with [D3]L-leucine, and metabolic rates were calculated by computer modeling. The overall effect of oral estrogen therapy on apoB metabolism was to accelerate the fractional catabolic rates of all particles studied and production rates of all except IDL. For light LDL (density = 1.019 1.036 g/mL), estrogen increased the mean fractional catabolic rate by 63% from 0.59 to 0.96 pools/day (P = 0.02), whereas the production rate increased by a lesser amount (42%) from 575 to 817 mg/day (P = 0.10). These metabolic changes reduced light LDL cholesterol and apoB concentrations by 26% (P = 0.005) and 19% (P = 0.03), respectively. In contrast, dense LDL (density = 1.036-1.063 g/mL) cholesterol and apoB concentrations were unchanged by the intervention, as both the apoB fractional catabolic rate and production rate were significantly increased by similar amounts, 39% (from 0.41 to 0.57 pools/day, P = 0.01) and 38% (from 434 to 601 mg/day; P = 0.003), respectively. Estrogen decreased the predominant LDL peak particle size from 273 to 268 A (P = 0.04). Thus, estrogen therapy increases the clearance of both light and dense LDL, counteracting increases in production rates. The reduced plasma residence times of light and dense LDL both may be antiatherogenic, even though, for dense LDL, the concentration did not change. PMID- 9398696 TI - Mutations and disorders involving the thyroid iodide transporter--the next wave in thyroid diseases. PMID- 9398697 TI - A homozygous missense mutation of the sodium/iodide symporter gene causing iodide transport defect. AB - Iodide transport defect is a disorder characterized by an inability of the thyroid to maintain an iodide concentration difference between the plasma and the thyroid. The recent cloning of the sodium/iodide symporter (NIS) gene enabled us to characterize the NIS gene in this disorder. We identified a homozygous missense mutation of A-->C at nucleotide +1060 in NIS complementary DNA in a male patient who was born from consanguineous marriage, had a huge goiter, and lacked the ability to accumulate iodide but was essentially euthyroid. The mutation results in an amino acid replacement of Thr354-->Pro in the middle of the ninth transmembrane domain. COS-7 cells transfected with the mutant NIS complementary DNA showed markedly decreased iodide uptake, confirming that this mutation was the direct cause of the disorder in the patient. Northern analysis of thyroid ribonucleic acid revealed that NIS messenger ribonucleic acid level was markedly increased (> 100-fold) compared with that in the normal thyroid, suggesting possible compensation by overexpression. PMID- 9398698 TI - A 37-year-old female with a shoulder mass and hypertensive crisis. PMID- 9398699 TI - Serum inhibin B in healthy pubertal and adolescent boys: relation to age, stage of puberty, and follicle-stimulating hormone, luteinizing hormone, testosterone, and estradiol levels. AB - Inhibin B levels were measured in serum from 400 healthy Danish prepubertal, pubertal, and adolescent males, aged 6-20 yr, in a cross-sectional study using a recently developed immunoassay that is specific for inhibin B, the physiologically important inhibin form in men. In addition, serum levels of FSH, LH, testosterone, and estradiol levels were measured. Serum levels of inhibin B, FSH, LH, testosterone, and estradiol all increased significantly between stages I and II of puberty. From stage II of puberty the inhibin B level was relatively constant, whereas the FSH level continued to increase between stages II and III. From stage III of puberty the FSH level was also relatively constant, although there was a nonsignificant trend of slightly decreased FSH levels at pubertal stage V compared to stage IV. The levels of serum LH, testosterone, and estradiol increased progressively throughout puberty. In prepubertal boys younger than 9 yr, there were no correlation between inhibin B and the other three hormones. In prepubertal boys older than 9 yr, a significant positive correlation was observed between inhibin B and FSH, LH, and testosterone. However, at this pubertal stage, each hormone correlated strongly with age, and when the effect of age was taken into account, only the partial correlation between inhibin B and LH/testosterone remained statistically significant. At stage II of puberty, the positive partial correlation between inhibin B and LH/testosterone was still present. At stage III of puberty, an negative partial correlation between inhibin B and FSH, LH, and estradiol was present, whereas no correlation between inhibin B and testosterone could be observed from stage III onward. The negative correlation between inhibin B and FSH persisted from stage III of puberty onward, whereas the correlation between inhibin B and LH and between inhibin B and estradiol was nonsignificant at stages IV and V of puberty. In conclusion, in boys, serum inhibin B levels increase early in puberty; by pubertal stage II the adult level of inhibin B has been reached. The correlation of inhibin B to FSH, LH, and testosterone changes during pubertal development. Early puberty is characterized by a positive correlation between inhibin B and LH/testosterone, but no correlation to FSH. Late puberty (from stage III) is characterized by a negative correlation between inhibin B and FSH (which is maintained in adult men), a diminishing negative correlation between inhibin B and LH, and no correlation between inhibin B and testosterone, suggesting that developmental and maturational processes in the hypothalamic-pituitary-gonadal axis take place, leading to the establishment of the closed loop feedback regulation system operating in adult men. The positive correlation between inhibin B and LH/ testosterone at the time when serum inhibin B levels rise early in puberty suggests that Leydig cell factors may play an important role in the maturation and stimulation of Sertoli cells in the beginning of pubertal development. PMID- 9398700 TI - Use of insulin-like growth factor I (IGF-I) and IGF-binding protein measurements to monitor feeding of premature infants. AB - To determine whether peptides of the insulin-like growth factor (IGF) system might be useful indicators of nutritional adequacy in premature infants, we studied 50 premature (25-34 weeks gestation) infants prospectively to define the relationship between nutrient intake and serum concentrations of IGF-I, IGF binding protein-2 (IGFBP-2), and IGFBP-3. Each infant was monitored for at least 2 weeks. Nutrient intake was quantified from daily logs; weight was determined daily, and measurements of IGF-I, IGFBP-2, and IGFBP-3 in serum were made twice weekly. Serum IGF-I correlated strongly with length of gestation, increasing 4.03 +/- 0.95 ng/mL for each additional week of gestation (P < 0.0001) and 0.36 +/- 0.07 ng/mL day each day since birth (P < 0.0001). A higher intake of calories increased IGF-I by 0.07 +/- 0.01 ng/mL for each calorie per kg ingested over the previous 3 days (P < 0.0001). IGF-I increased quadratically as protein intake increased. For each change of 1% in calories as protein squared, IGF-I increased 0.36 +/- 0.11 ng/mL (P < 0.0001). Serum IGFBP-3 concentrations also correlated with length of gestation, increasing 25.06 +/- 11.83 micrograms/L.wk (P = 0.035) and 4.14 +/- 1.33 micrograms/.day since birth (P = 0.003). Unlike IGF-I, variation in the amount of protein supplied did not change IGFBP-3. As calorie intake increased, IGFBP-3 increased by 0.54 +/- 0.17 microgram/L for each calorie per kg consumed over the previous 3 days (P = 0.0015). In contrast to IGF-I and IGFBP-3, IGFBP-2 declined as the length of gestation increased (56.12 +/- 16.92 ng/mL.week; P = 0.001) and with each additional day of life (7.57 +/- 2.44 ng/mL.day; P = 0.003). Dietary protein, the predominant regulator of IGFBP-2, caused a decrease of 33.22 +/- 9.00 ng/mL with each percent increase in dietary calories as protein (P < 0.0003). Calorie intake had less effect on IGFBP-2 than protein intake. These results indicate that each of the three peptides studied is regulated in premature infants by nutritional intake, and that their regulatory patterns are qualitatively similar to those observed in older individuals. Measurements of these peptides in premature infants may be useful indicators of nutritional status and adequacy of nutrient intake. PMID- 9398701 TI - Effects of luteinizing hormone-releasing hormone analog-induced pubertal delay in growth hormone (GH)-deficient children treated with GH: preliminary results. AB - To study the effect of delaying epiphyseal fusion on the growth of GH-deficient children, we studied 14 pubertal, treatment naive, GH-deficient patients (6 girls and 8 boys) in a prospective, randomized, placebo-controlled trial. Chronological age was 14.5 +/- 0.5 yr, and bone age was 11.6 +/- 0.3 yr (mean +/- SEM) at the beginning of the study. Patients were assigned randomly to receive GH and LH releasing hormone (LHRH) analog (n = 8) or GH and placebo (n = 6) during 3 yr, with planned continuation of GH treatment until epiphyseal fusion. Patients were measured with a stadiometer and had serum LHRH tests, serum testosterone (boys), serum estradiol (girls), and bone age performed every 6 months. Patients treated with GH and LHRH analog showed a clear suppression of their pituitary-gonadal axis and a marked delay in bone age progression. We observed a greater gain in height prediction in these patients than in the patients treated with GH and placebo after 3 yr of treatment (mean +/- SEM, 14.0 +/- 1.6 vs. 8.0 +/- 2.4 cm; P < 0.05). These preliminary findings suggest that delaying epiphyseal fusion with LHRH analog in pubertal GH-deficient children treated with GH increases height prediction and may increase final height compared to treatment with GH alone. PMID- 9398702 TI - Diabetes during pregnancy does not alter whole body bone mineral content in infants. AB - A previous study using single photon absorptiometry has reported low bone mineral density of the radius in infants of diabetic mothers. The aim of this study was to assess by dual x-ray absorptiometry the whole body bone mineral content (WbBMC) and the body composition of 40 infants of diabetic mothers at birth (mean gestational age +/- SD, 37.5 +/- 1.3 weeks; mean birth weight +/- SD, 3815 +/- 641 g). WbBMC was not correlated with gestational age, but was well correlated with birth weight (r = 0.73; P = 0.0001) and also with fat mass (r = 0.87; P = 0.0001) and lean mass (r = 0.42; P = 0.008). The z-scores +/- SD adjusted for weight for WbBMC and fat mass were significantly increased (1.3 +/- 0.9 and 2.6 +/- 1.3, respectively (P < 0.0001), but were not significantly influenced either by in utero growth or by the type of the diabetes mellitus of the mother. Bone mineralization and fat mass studied by whole body dual x-ray absorptiometry are increased at birth in these infants compared with reference curves. PMID- 9398703 TI - Characterization of monoclonal thyroid-stimulating and thyrotropin binding inhibiting autoantibodies from a Hashimoto's patient whose children had intrauterine and neonatal thyroid disease. AB - A multiplicity of TSH receptor autoantibodies (TSHRAbs) have been characterized after subcloning heterohybridomas produced from the lymphocytes of a patient who has Hashimoto's thyroiditis and had three children with intrauterine or neonatal hyperthyroidism. Twelve clones produced stimulating TSHRAbs that increased cAMP levels and iodide uptake in rat FRTL-5 thyroid cells and increased cAMP levels in Chinese hamster ovary (CHO) cells transfected with the human TSHR; like 95% of Graves' stimulating TSHRAbs, all 12 have their functional epitope on the N terminus of the TSHR extracellular domain, requiring residues 90-165 for activity. All 12 bind to human thyroid membranes in the absence, but not the presence, of TSH, but are only weak inhibitors of TSH binding in assays measuring TSH binding-inhibiting Igs (TBIIs). In contrast, 8 different clones produced TSHRAbs that did not increase cAMP levels, but, instead, exhibited significant TBII activity. Four inhibited the ability of TSH or a stimulating TSHRAb to increase cAMP levels and had their functional epitope on the C-terminal portion of the TSHR external domain, residues 261-370, mimicking the properties of blocking TSHRAbs that cause hypothyroidism in patients with idiopathic myxedema. The 4 other TBIIs inhibited the ability of TSH, but not that of a stimulating TSHRAb, to increase cAMP levels, like TBIIs in Graves' patients. The functional epitope for 3 of these Graves'-like TBIIs was residues 90-165; the functional epitope for the fourth was residues 24-89. The fourth also increased arachidonic acid release and inositol phosphate levels in FRTL-5 thyroid cells and exhibited conversion activity, i.e. the ability to increase cAMP levels in the presence of an anti-human IgG. Thus, this TBII exhibited signal transduction activity, unlike the other 3 Graves'-like TBIIs. The patient, therefore, has stimulating TSHRAbs and 3 different types of TBIIs, each with different functional properties and different epitopes on the TSHR. PMID- 9398704 TI - Acute effects of estradiol infusion and naloxone on luteinizing hormone secretion in pubertal boys. AB - We have shown previously in pubertal boys that testosterone (T) suppresses the nocturnal augmentation of luteinizing hormone (LH) secretion principally by decreasing LH pulse frequency. As T can be aromatised to estradiol (E2), and E2 effects on LH secretory dynamics may be separate from those of T, we examined the effects of acute E2 infusion on LH secretion in pubertal boys. Opioid receptor blockade has been reported to increase LH secretion after estradiol suppression in adult men, so we also examined whether naloxone might augment LH secretion during E2 treatment in pubertal boys. Starting at 1000 h, eight pubertal boys were given a 33 h saline infusion, followed 1 week later by an E2 infusion at 4.6 nmol/m2/h. During both infusions, four iv boluses of saline were given hourly beginning at 1200 h on the first day, and four naloxone iv boluses, 0.1 mg/kg each, were given hourly beginning at 1200 h on the second day. Blood was obtained every 15 min for LH, and every 60 min for T and E2, from 1200 h until the end of the infusion. Pituitary responsiveness to gonadotropin-releasing hormone (GnRH) was assessed after both infusions by iv administration of 250 ng/kg synthetic GnRH. Estradiol infusion increased the mean plasma E2 concentration from 23 +/- 4 to 46 +/- 6 pmol/L (P < 0.01) and suppressed mean plasma T from 4.9 +/- 1.4 to 3.0 +/- 3.5 nmol/L (saline vs. E2 infusion, P < 0.05). The overall mean LH was suppressed by E2 infusion from 3.7 +/- 0.5 to 2.2 +/- 0.4 IU/L (saline vs. E2 infusion, P < 0.01). LH pulse frequency was suppressed by 50%, whereas mean LH pulse amplitude was not different between saline and E2 infusions. Administration of naloxone did not alter the mean LH, LH pulse frequency, or amplitude during either saline or E2 infusions. Pituitary responsiveness to exogenous GnRH was similar during both infusions. These studies indicate that E2 produces its negative feedback in pubertal boys principally by suppression of LH pulse frequency, and naloxone does not reverse these suppressive effects. Thus E2 suppression of LH secretion is mediated by a decrease of hypothalamic GnRH secretion that is independent of endogenous opioid pathways. PMID- 9398705 TI - Seasonal variation in glucocorticoid activity in healthy men. AB - Many endocrine systems are subject to seasonal variation. However, studies of the hypothalamic-pituitary-adrenal axis in man have been limited to patients with psychiatric illness with conflicting results. We studied 105 healthy men, age 24 33 yr, during a 15-month period that included two winters. We measured cortisol and its metabolites by gas chromatography/mass spectrometry in plasma and urine and the intensity of dermal blanching after overnight topical application of beclomethasone dipropionate. There were no differences between subjects studied during the two winter periods, but marked differences between subjects studied in winter and summer. In winter, 0900-h plasma cortisol concentrations were higher (73 +/- 10 ng/mL, n = 41 vs. 35 +/- 4, n = 25 in summer; P < 0.01), total cortisol metabolite excretion was lower (678 +/- 67 micrograms/mmol creatinine vs. 900 +/- 98; P < 0.05), the ratio of metabolites of cortisol to those of cortisone was higher (3.0 +/- 0.2 vs. 2.1 +/- 0.1; P < 0.01), and dermal glucocorticoid sensitivity was higher (7.2 +/- 0.4 arbitrary units vs. 5.6 +/- 0.5; P < 0.02). Although blood pressure and fasting insulin/glucose relationships were not measurably different between seasons, these correlated with dermal vasoconstriction and cortisol metabolite excretion rate. We conclude that plasma cortisol and tissue sensitivity to glucocorticoids are higher in winter, but cortisol production rate is reduced. This could be explained by a reduction in cortisol clearance rate: urinary free cortisol/cortisone ratios were not different but A-ring-reduced metabolites of cortisol were higher in winter, suggesting that conversion of cortisone to cortisol by hepatic 11 beta hydroxysteroid dehydrogenase 1 is enhanced. It is an intriguing possibility that increased glucocorticoid activity contributes to the increased prevalence of disease during the winter. PMID- 9398706 TI - Thyroid nodules in the follow-up of irradiated individuals: comparison of thyroid ultrasound with scanning and palpation. AB - In 1974 we began a prospective study of a cohort of 4296 individuals exposed to therapeutic head and neck irradiation during childhood for benign conditions. To define the role of thyroid ultrasonography in following irradiated individuals, we studied a subgroup of 54 individuals. They all had been screened between 1974 1976 and had normal thyroid scans and no palpable nodules at that time. Thyroid ultrasonography, thyroid scanning, physical examination, and serum thyroglobulin measurements were performed. One or more discrete ultrasound-detected nodules were present in 47 of 54 (87%) subjects. There were a total of 157 nodules, 40 of which were 1.0 cm or larger in largest dimension. These 40 nodules occurred in 28 (52%) of the subjects. Thirty (75%) of these 1.0-cm or larger nodules matched discrete areas of diminished uptake on corresponding thyroid scans. The 10 that did not match (false negative scans for > or = 1.0-cm nodules) were the only nodules of this size in 7 subjects. Of 11 nodules 1.5 cm or larger, only 5 were palpable. Serum thyroglobulin correlated to the number (P = 0.04; r2 = 0.10), but not the volume of the thyroid nodules (P = 0.07; r2 = 0.08). We conclude that thyroid nodules are continuing to occur and are exceedingly common in this irradiated cohort of individuals. The results confirm that thyroid ultrasonography is more sensitive than physical examination and scanning. However, thyroid ultrasound is so sensitive and nodules so prevalent that great caution is needed in interpreting the results. PMID- 9398707 TI - Fetal plasma hypoxanthine level in growth-retarded fetuses before labor. AB - Hypoxanthine is one of the purine nucleotides and is presumed to accumulate during hypoxia and acidemia. It remains uncertain, however, whether plasma hypoxanthine concentration is a useful indicator of fetal asphyxia; and its relationship to other markers of fetal physiologic state is not clearly defined. The aim of this study was to evaluate whether the level of fetal plasma hypoxanthine is correlated with fetal hypoxia and acidosis in growth-retarded fetuses before the onset of labor. Cordocentesis was performed in 34 growth retarded fetuses at 31-35 weeks' gestation for the measurement of umbilical venous plasma concentrations of hypoxanthine, hemoglobin and lactate concentrations, blood gases, and base deficit. Umbilical venous plasma hypoxanthine concentration was found to be increased significantly, in parallel with the degree of acidosis (r = -0.74, P < 0.05) and base deficit (r = -0.41, P < 0.05), but not to bear a significant relationship to the degree of hypoxemia or other measured variables. We conclude that increases in the plasma concentration of hypoxanthine may reflect an impaired physiological state in growth-retarded fetuses before labor. PMID- 9398708 TI - Decreased hypothalamic thyrotropin-releasing hormone gene expression in patients with nonthyroidal illness. AB - Changes in hypothalamus-pituitary-thyroid function occur in patients with a variety of illnesses and are referred to as the euthyroid sick syndrome or nonthyroidal illness (NTI). In NTI, serum concentrations of T3 decrease to low, or even undetectable, levels without giving rise to elevated concentrations of TSH. We hypothesized that decreased activity of TRH-producing cells in the paraventricular nucleus (PVN) contributes to the persistence of low TSH levels. To test this hypothesis, we collected a series of formalin-fixed, paraffin embedded hypothalami of patients whose plasma concentrations of T3, T4, and TSH had been measured in a blood sample taken less than 24 h before death. Quantitative TRH messenger RNA in situ hybridization (intraassay coefficient of variation: 13%) was performed in the PVN. Total TRH messenger RNA in the PVN showed a positive correlation with serum T3 (r = 0.66; P < 0.05) and with logTSH (r = 0.64; P < 0.05), but not with T4 (r = -0.02; P = 0.95). This is the first study to correlate premortem serum concentrations of thyroid hormones with postmortem gene expression of identified neurons in the human hypothalamus. The results suggest an important role for TRH cells in the pathogenesis of NTI. PMID- 9398709 TI - Effect of obesity on the response to insulin therapy in noninsulin-dependent diabetes mellitus. AB - An initial improvement in glycemic control is often followed by gradual deterioration of glycemia during insulin treatment of patients with noninsulin dependent diabetes mellitus (NIDDM). We examined the causes of such worsening in a 12-month follow-up analysis of 100 insulin-treated NIDDM patients in the Finnish Multicenter Insulin Therapy Study who were treated with either combination therapy with insulin or insulin alone. In the entire study group, glycemic control averaged 9.7 +/- 0.2% at 0 months and 8.0 +/- 0.1%, 8.0 +/- 0.1%, 8.2 +/- 0.1%, and 8.5 +/- 0.2% at 3, 6, 9, and 12 months (P < 0.001 for each time point vs. 0 months). Glycemic control at 12 months was significantly worse than that at 3 (P < 0.001), 6 (P < 0.001), and 9 months (P < 0.02). Baseline body mass index was the most significant predictor of deterioration in glycemic control. During 1 yr, hemoglobin A1c decreased almost 3-fold more (by 1.7 +/- 0.2%; P < 0.001 vs. 0 months) in patients whose baseline weight was below the mean baseline body mass index of 28.1 kg/m2 (nonobese patients) than in those whose weight exceeded 28.1 kg/m2 (obese patients; 0.5 +/- 0.2%; P = NS vs. 0 months; P < 0.01 vs. obese patients). Glycemic control improved similarly over 1 yr in the nonobese subjects and deteriorated similarly in the obese patients regardless of their treatment regimen. Insulin doses, per body weight, were similar in the nonobese and obese patients. The nonobese patients consistently gained less weight during 12 months of combination therapy with insulin (3.5 +/- 0.6 kg at 12 months) than during insulin therapy alone (5.1 +/- 0.6 kg; P < 0.05). The treatment regimen did not influence weight gain in the obese group, who gained 4.4 +/- 1.0 kg during combination therapy with insulin and 4.5 +/- 1.1 kg during insulin therapy alone. We reached the following conclusions: 1) after an initial good response, glycemic control deteriorates more in obese than in nonobese patients with NIDDM; 2) in obese patients, weight gain per se cannot explain the poor glycemic response to combination or insulin therapy, but it may induce a disproportionately large increase in insulin requirements because of greater insulin resistance in the obese than in the nonobese; 3) in nonobese patients, glycemic control improves equally during 1 yr with combination therapy with insulin and insulin alone, but combination therapy with insulin is associated with less weight gain than treatment with insulin alone; 4) weight gain appears harmful, as it is associated with increases in blood pressure and low density lipoprotein cholesterol. PMID- 9398710 TI - A novel cyclic adenosine monophosphate analog induces hypercalcemia via production of 1,25-dihydroxyvitamin D in patients with solid tumors. AB - The treatment of cancer patients with conventional chemotherapy is sometimes associated with severe systemic toxicity and only a minimal survival benefit. Because of this, new less toxic and more efficacious treatments have been sought. 8-Chloro-cAMP (8-Cl-cAMP) is one of a new generation of anticancer drugs that act at the level of signal transduction. In preclinical models, 8-Cl-cAMP modulates protein kinase A (PKA) leading to growth inhibition and increased differentiation of cancer cells. 8-Cl-cAMP was given to 16 patients with advanced cancer as an infusion via an indwelling subclavian venous catheter. We showed that 8-Cl-cAMP had a parathyroid hormone-like effect leading to increased synthesis of renal 1,25-dihydroxyvitamin D [up to 14 times the baseline value, median 3.6 times; P = 0.00001 (Student's paired t test)]. This produced the dose-limiting toxicity of reversible hypercalcemia that could not be controlled by the administration of either pamidronate or dexamethasone. The treatment was otherwise well tolerated, and other cAMP-dependent pathways (cortisol and TSH) were not affected, emphasizing the marked differences between organs in their sensitivity to this cAMP analog. Our results have shown that 8-Cl-cAMP is biologically active, and it is feasible that if the hypercalcemia can be controlled, then this drug may have a role as a single agent, or as a short infusion between cycles of chemotherapy. PMID- 9398711 TI - Randomized, double blind, placebo-controlled trial of low dose iodide in endemic goiter. AB - Iodine (I) is essential for normal thyroid function, and the majority of subjects tolerate a wide range of dietary levels. However, a subset of individuals upon exposure to normal or elevated levels of I develop thyroid dysfunction and autoimmunity. In this double blind trial, we evaluated efficacy and tolerability of low dose I in adults with euthyroid, diffuse, endemic goiter. Sixty-two subjects were randomly assigned I (0.2 mg/day) or placebo for 12 months. After termination of therapy, both groups were followed for a further 6 months. Thyroid sonography and determinations of thyroid-related hormones, urinary I excretion per 24 h, and thyroid antibodies were carried out at baseline and at 3, 6, 9, 12, 15, and 18 months. Markedly elevated urinary I values were found during therapy in subjects receiving I (32 at baseline vs. 213 micrograms/24 h at 12 months; P = 0.0001) compared to placebo (34 and 33 micrograms/24 h, respectively; P < 0.0001 vs. I). I substantially reduced thyroid volume (29 vs. 18 mL at 12 months; -38%; P = 0.0001), and at 18 months, the therapeutic effect was sustained. In the placebo group, no significant changes were observed. High microsomal and thyroglobulin autoantibody titers were present in 3 of 31 (9.7%) subjects receiving I, and I-induced hypo- and hyperthyroidism developed in 2 and 1, respectively. Fine needle biopsy revealed marked lymphocytic infiltration in all 3 cases. After withdrawal of I, thyroid dysfunctions spontaneously remitted, and antibody titers as well as lymphocytic infiltration decreased markedly. Follow-up of these 3 subjects for an additional 2 yr showed normalization of antibody titers in 2. Thus, among subjects with endemic goiter, low dose I successfully normalized thyroid volume and body I supplementation; nevertheless, reversible I induced thyroid dysfunctions and autoimmunity were observed in nearly 10% of the subjects. PMID- 9398712 TI - A new compound heterozygous mutation in the 11 beta-hydroxysteroid dehydrogenase type 2 gene in a case of apparent mineralocorticoid excess. AB - Apparent mineralocorticoid excess (AME) characterized by early-onset hypertension and hypokalemia is due to congenital deficiency of 11 beta-hydroxysteroid dehydrogenase (11 beta HSD). Two isoforms of human 11 beta HSD are known, and the type 2 isoform (11 beta HSD2) has been recently shown to be responsible for AME. In this study we have analyzed the 11 beta HSD2 gene of a Japanese patient with AME. PCR amplification and subsequent nucleotide sequencing of the 11 beta HSD2 gene from the patient and his family members revealed that the patient has a compound heterozygous mutation of this gene. In 1 allele, an undescribed single nucleotide transition in codon 208 in exon 3 resulted in a substitution of arginine to histidine (CGC to CAC: R208H). In the other allele, a deletion of 3 nucleotides in codons 337-338 in exon 5 resulted in a substitution of arginine to histidine and a deletion of tyrosine residue (CGCTAT to CAT: R337H, delta Y338), which has been previously shown to abolish 11 beta HSD2 enzyme activity. A chloramphenicol acetyltransferase assay-based expression study involving the mineralocorticoid receptor indicated that the novel R208H mutation eliminates the enzymatic activity of 11 beta HSD2. From the genetic analysis of 50 healthy subjects, the novel R208H mutation was unlikely to be due to polymorphism. Together, these results indicate that this patient is a compound heterozygote for the mutation in the 11 beta HSD2 gene (R208H and R337H, delta Y338) and that these mutations inactivate the 11 beta HSD2 function and give rise to clinically manifest AME. PMID- 9398713 TI - Inhibin B as a serum marker of spermatogenesis: correlation to differences in sperm concentration and follicle-stimulating hormone levels. A study of 349 Danish men. AB - Recent studies have focused on reproductive health of men in the general population. However, semen samples are difficult to obtain within sampling frames that allow comparisons. Blood samples are easier to obtain than ejaculates. Therefore, serum biomarkers of spermatogenesis are of major interest for population studies. FSH has previously been used as a marker of spermatogenesis, although it is also influenced by the hypothalamus. Serum inhibin B was recently suggested as a possible, more direct serum marker of spermatogenesis in men with testicular disorders. In a Danish nationwide collaborative study, we found an unexpected difference in semen concentration between two groups of men recruited from two different centres. We, therefore, analyzed reproductive hormones in blood, including inhibin B, to test whether the observed difference in semen concentration was reflected in the reproductive hormones. From 1992 to 1995, a total of 430 men, 20-35 yr old, who lived with a partner and who had not previously attempted to achieve a pregnancy, were recruited. The couples were enrolled into the study in one of two centres (centre A, n = 231; and centre B, n = 199) when they discontinued birth control. At enrollment, they provided a semen sample (n = 419), and a blood sample was drawn (n = 349). The semen analysis was performed in accordance with the WHO 1992 guidelines, and interlaboratory differences were tested. Inhibin B was measured in an enzyme immunometric assay, which has previously been described. All blood samples were analyzed in the same laboratory. Median sperm concentration and the percentage of morphologically normal spermatozoa were significantly higher among men from centre A (56.0 mill/mL and 42.5%), compared with men from centre B (44.8 mill/mL and 39%). Men from centre B had a significantly higher median FSH (3.42 IU/L) and a lower inhibin B (186 pg/mL) than men from centre A (3.21 IU/L and 209 pg/mL). The differences persisted after control for potentially confounding variables. A significant correlation was found between the cubic root-transformed serum FSH and inhibin B levels (r = -0.61, P < 0.001), between the cubic root-transformed serum FSH and sperm concentration (r = -0.40, P < 0.001), and between the cubic root-transformed inhibin B and sperm concentration (r = 0.38, P < 0.001). The predictive power of detecting sperm counts below 20 mill/mL among men who's inhibin B and FSH both were below 80 pg/mL and above 10 IU/L, respectively, was 100%. The unexpected significant difference in semen concentration between two groups of normal Danish men was probably caused by differences in sampling procedures in the two centres where the men were recruited, rather than geographical differences. However, similar differences in serum levels of inhibin B and FSH between centres were found. These findings suggest that a real difference in spermatogenic potential between the two groups of men existed. We suggest that serum inhibin B, in future population studies on male reproductive health, may serve as a new marker of spermatogenesis, in addition to sperm concentration and serum FSH. PMID- 9398714 TI - Metabolism of progesterone by human lymphocytes: production of neuroactive steroids. AB - Although it has long been recognized that lymphocytes have the capacity to reduce cortisol at the C3, C5, and C20 positions, the specificity and the physiological variation of these reactions have received little attention. We have shown that such reactions also occur with progesterone. Lymphocytes were isolated from whole blood using Percoll density gradient centrifugation. The cells were incubated for 20 h with tritiated progesterone as radioactive tracer. After extractions into ethyl acetate, the residue was subjected to high performance liquid chromatography, and the radioactivities of the separated compounds were determined. Without cells, 95-97% of the tracer added was recovered in the progesterone peak, while in the presence of 4 x 10(6) lymphocytes, this was reduced to 45-90%. The metabolites obtained included at least 10 different compounds, including those corresponding in their retention times to the neuroactive 5 alpha and 5 beta dihydroprogesterones and their 3 alpha- and 3 beta tetrahydroprogesterone derivatives. The conversion decreased with the addition of other steroids such as testosterone, cortisol, and corticosterone, suggesting that these steroids are metabolized by the same enzymes. When the cells from two pregnant patients were combined and incubated with tracer, and with and without nonradioactive progesterone, no peaks were detected by two progesterone radioimmunoassays in the absence of added nonradioactive progesterone, while in its presence three peaks corresponding to 5 alpha-dihydroprogesterone, 3 alpha hydroxy-5 alpha-pregnane-20-dione and 3 beta-hydroxy-5 alpha-pregnane-20-dione eluted before the P peak. Their identities were confirmed using the two different progesterone radioassays that cross-reacted with these metabolites. The highest mean conversion (44.7% +/- 3.2 SE) was found with the lymphocytes of pregnant women and with that of one lactating woman (50%). Conversions by lymphocytes of women in the follicular phase (29.3% +/- 1.3 SE) were significantly lower than those in pregnancy (P = 0.014) but did not differ significantly (P > or = 0.05) from those of women in the luteal phase (22.2% +/- 3.4 SE), those of postmenopausal women (23.5% +/- 4.9 SE), or of men (22.5% +/- 2.4 SE). Lymphocytes appear to provide a hitherto unrecognized but possibly important source of neuroactive steroids. PMID- 9398715 TI - Studies of genetic variability of the uncoupling protein 1 gene in Caucasian subjects with juvenile-onset obesity. AB - Our objective was to investigate whether genetic variants of the uncoupling protein 1 (UCP1) gene are associated with juvenile-onset obesity or alterations in weight gain and insulin sensitivity in young healthy Caucasians. Single-strand conformation polymorphism and heteroduplex analysis of the coding region of the UCP1 gene was performed in 56 subjects randomly selected at the draft board examination from a cohort of 156 males with juvenile-onset obesity. Association studies of amino acid variants were undertaken in the cohort of males with juvenile-onset obesity, a cohort of 205 randomly selected control males, and a subgroup of this cohort comprising 76 lean subjects. Genetic variants of the coding region as well as a previously described a-->g nucleotide polymorphism of the 5'-flanking region of the UCP1 gene were examined for associations with accelerated weight gain or reduced sensitivity to insulin in a cohort of 380 young healthy Caucasians. The mutational analysis revealed five nucleotide substitutions that changed the sequence of UCP1, Arg/Trp40, Ala/Thr64, Val/Met137, Met/Leu229, and Lys/Asn257 and two nucleotide substitutions in the nontranslated region of exon 1. Among subjects with juvenile-onset obesity, the allelic frequencies of Ala/Thr64 and Met/Leu229 were both 8.2% (95% confidence interval: 5.1-11.3%) vs. 8.8% (6.0-11.6%) and 8.1% (5.3-10.9%), respectively, in the cohort of randomly selected control subjects. Among lean control subjects, the allelic frequencies of the polymorphisms were 8.2% (3.7-12.7%) and 5.6% (1.9 9.3%), respectively. In the cohort of young healthy subjects, measurements of obesity and insulin sensitivity did not differ between carriers of the Ala/Thr64 and Met/Leu229 variants and wild-type carriers. The Val/Met137 and Lys/Asn257 mutations were each found in one subject with juvenile-onset obesity, and the Arg/Trp40 mutation was found in two obese subjects and in one control subject. The allelic frequency of the nucleotide polymorphism of the 5'-flanking region of the UCP1 gene was 25.3% (22.2-28.4%) in the cohort of 380 young Danes. There were no differences in body mass index, fat mass, waist-to-hip ratio, or weight gain during childhood or adolescence between carriers and noncarriers of this nucleotide variant. Although we cannot exclude an effect of the rare mutations in the UCP1 gene on susceptibility to juvenile-onset obesity, genetic variation of the coding region of the UCP1 gene is not a common factor contributing to obesity in Caucasian subjects of Danish ancestry. PMID- 9398716 TI - Lean women with polycystic ovary syndrome respond to insulin reduction with decreases in ovarian P450c17 alpha activity and serum androgens. AB - It is unknown whether hyperinsulinemia plays a role in the pathogenesis of polycystic ovary syndrome (PCOS) in normal weight or thin women. Evidence indicates that these women are insulin resistant and hyperinsulinemic, and this study was conducted to test the hypothesis that hyperinsulinemia stimulates ovarian cytochrome P450c17 alpha activity in nonobese women with PCOS, thereby increasing serum androgen concentrations. We assessed ovarian P450c17 alpha activity (by measuring the response of 17 alpha-hydroxyprogesterone to a GnRH agonist), fasting serum steroids, and oral glucose tolerance before and after oral administration of either metformin (500 mg) or placebo three times daily for 4-6 weeks in 31 nonobese women with PCOS. In the 19 women given metformin, the mean (+/- SE) area under the serum insulin curve after oral glucose administration decreased from 44 +/- 5 to 24 +/- 3 nmol/L.min (P = 0.003). Basal serum 17 alpha-hydroxyprogesterone decreased from 3.4 +/- 0.3 to 2.5 +/- 0.4 nmol/L (P = 0.05), and GnRH-stimulated peak serum 17 alpha-hydroxyprogesterone decreased from 12.2 +/- 1.6 to 7.5 +/- 0.7 nmol/L (P = 0.005). Serum 17 alpha hydroxyprogesterone values did not change in the placebo group. In the metformin group, serum free testosterone decreased by 70% from 18.2 +/- 3.1 to 5.5 +/- 0.7 pmol/L (P < 0.001), and serum sex hormone-binding globulin increased from 84 +/- 6 to 134 +/- 15 nmol/L (P = 0.002). None of these values changed in the placebo group. These findings suggest that hyperinsulinemia stimulates ovarian P450c17 alpha activity in nonobese women with PCOS. They also indicate that decreasing serum insulin with metformin reduces ovarian cytochrome P450c17 alpha activity and ameliorates the hyperandrogenism of these women. PMID- 9398717 TI - Tumor necrosis factor increases serum leptin levels in humans. AB - Leptin is a pleiotropic hormone believed to regulate body weight. Its function in wasting during inflammatory disease in humans is unknown. We studied the effect of repeated tumor necrosis factor (TNF) infusion on serum leptin levels in six patients with solid tumors. TNF infusion on day 1 resulted in an increase in serum leptin levels from 3.1 (SEM +/- 0.28) ng/mL to 5.2 (SEM +/- 0.6) ng/mL after 12 h (P < 0.001). The serum levels returned to baseline within 24 h. Similar results were obtained when TNF was infused on subsequent days. The study shows that leptin serum levels are under control of TNF. PMID- 9398718 TI - Insulin does not stimulate protein synthesis acutely in prepubertal children with insulin-dependent diabetes mellitus. AB - Insulin treatment in adult type I diabetic patients decreases protein loss primarily by inhibiting protein breakdown without stimulating protein synthesis. In young growing rodents, insulin treatment has been reported to stimulate protein synthesis. We examined whether insulin stimulates protein synthesis in normally growing prepubertal children with insulin-dependent diabetes mellitus. Five prepubertal children with insulin-dependent diabetes mellitus (aged 8.6 11.25 yr) were studied in the postabsorptive state on two occasions: once during insulin deprivation (I-; blood glucose, 325 +/- 67.8 mg/dL; mean +/- SD) and once during insulin administration for 4 h (I+; blood glucose, 96 +/- 23.6 mg/dL). Leucine kinetics were measured using a 4-h primed continuous infusion of L-[1 13C]leucine. Serum insulin concentrations were lower (I- vs. I+, 0.6 +/- 0.3 vs. 7.5 +/- 4.3 microU/mL; mean +/- SD; P = 0.02), whereas serum beta-hydroxy butyrate (I- vs. I+, 3.4 +/- 0.5 vs. 0.9 +/- 0.5 mg/dL; P < 0.001) and free fatty acid concentrations (I- vs. I+, 2.9 +/- 0.4 vs. 0.9 +/- 0.4 mEq/L; P < 0.001) were higher in the insulin-deprived state than during insulin administration. Leucine Ra, an index of protein breakdown (I- vs. I+, 200.5 +/- 23.4 vs. 167 +/- 17 mumol/kg.h; P = 0.008), and leucine oxidation (I- vs. I+, 56.5 +/- 20.7 vs. 29.6 +/- 9.3 mumol/kg.h; P = 0.03) were reduced by insulin treatment. Nonoxidative leucine disposal, an index of protein synthesis, was not affected by insulin treatment (I- vs. I+, 144 +/- 20.8 vs. 137.5 +/- 13.5 mumol/kg.h; P = 0.4). We conclude that the acute decline in net protein loss during insulin treatment in growing prepubertal children, like that in adults, is due primarily to an inhibition of protein breakdown without stimulation of protein synthesis. PMID- 9398719 TI - Increased disorderliness of basal insulin release, attenuated insulin secretory burst mass, and reduced ultradian rhythmicity of insulin secretion in older individuals. AB - Insulin is secreted in a pulsatile fashion. Rapid pulses are considered to be important for inhibiting hepatic glucose output, and ultradian pulses for stimulating peripheral glucose disposal. Aging is characterized by a progressive impairment in carbohydrate tolerance. We undertook the current studies to determine whether alterations in pulsatile insulin release accompany the age related changes in carbohydrate metabolism. Healthy young (n = 8; body mass index, 21 +/- 1 kg/m2; age, 24 +/- 1 yr) and old (n = 9; body mass index, 24 +/- 1 kg/m2; age, 77 +/- 2 yr) volunteers underwent two studies. In the first study, insulin was sampled every 1 min for 150 min, and pulse analysis was conducted using a recently validated multiparameter deconvolution technique. In the second study, insulin was sampled every 10 min for 600 min, and insulin release was evaluated by Cluster analysis. In the 150-min studies, insulin secretory burst mass (P < 0.05) and amplitude (P < 0.01) were reduced in the elderly. In addition, approximate entropy, a measure of irregularity or disorderliness of insulin release, was increased in the aged (P < 0.01). In the 600-min studies, interpulse interval was greater in the aged (P < 0.05), and burst number was less (P < 0.05). We conclude that normal aging is characterized by more disorderly insulin release, a reduction in the amplitude and mass of rapid insulin pulses, and a decreased frequency of ultradian pulses. Whether these alterations in insulin pulsatility contribute directly to the age-related changes in carbohydrate metabolism will require further study. PMID- 9398720 TI - Cytokine production by a new undifferentiated human thyroid carcinoma cell line, FB-1. AB - A human anaplastic thyroid cancer cell line FB-1, derived from a 68-yr-old woman who underwent surgery for anaplastic thyroid cancer, has been established. The spindlelike cells have been proliferating stably for more than 2 yr. Karyotype analysis shows many abnormalities and many marker chromosomes have been observed. Heterotransplant of FB-1 cells into severe combined immunodeficient mice has resulted in rapidly growing tumors classified as anaplastic carcinomas, although 50% have shown areas with a trabecular pattern. FB-1 cells failed to express messenger RNA for thyroglobulin; TSH-receptor; thyroperoxidase, and placental angiogenic growth factor. Conversely, PAX8 and thyroid transcription factor 1, whose expression is thyroid specific, was kept in an FB-1 cell line at a level comparable with that observed in normal thyroid tissue. In addition, the present cell line expressed high levels of messenger RNA for high-mobility group proteins (Y) and -C. The in vitro study revealed that FB-1 cells are able to produce high levels of interleukin (IL)-8 and medium amount of IL-6, whereas no release of IL 1-alpha, IL-1-beta, and IL-4 was observed. No modulation of cell proliferation and DNA synthesis in FB-1 cells has been observed after the addition of exogenous IL-6. PMID- 9398721 TI - Sporadic pheochromocytomas are rarely associated with germline mutations in the vhl tumor suppressor gene or the ret protooncogene. AB - Pheochromocytoma is a tumor that may occur sporadically or may be a manifestation of a hereditary disease, such as von Hippel-Lindau disease (VHL) and multiple endocrine neoplasia (MEN) type 2. As patients with VHL or MEN type 2 are at risk to develop multiple tumors, they must be distinguished from sporadic cases. We determined the incidence of VHL and MEN type 2 among 62 German patients diagnosed with pheochromocytoma without a history of a hereditary disease. Germline analyses of the vhl gene and exons 10, 11, and 13 of the ret protooncogene were performed by PCR, single strand conformation polymorphism, enzyme digestion, or sequencing. Two patients (3%) showed vhl mutations (95% confidence interval, 1 11%). One patient showed loss of the MspI restriction site at nucleotides 712/713. Another patient had a C/T change at an intronic site that was also detected in 2 of his offspring. No mutations were detected in the ret protooncogene (97.5% confidence interval, 0-6%). In Germany, most sporadic pheochromocytomas are not due to VHL or MEN type 2. Therefore, clinical work-up in patients with pheochromocytoma without signs of hereditary disease is not recommended. However, because the costs of genetic screening are relatively low, and each index case allows optimal care for family members, molecular testing might be cost-effective. PMID- 9398722 TI - Functional differentiation of placental syncytiotrophoblasts during baboon pregnancy: developmental expression of chorionic somatomammotropin messenger ribonucleic acid and protein levels. AB - The objective of the present study was to determine whether, in addition to the onset of chorionic somatomammotropin (CS) production previously shown to result from the morphological differentiation of cytotrophoblasts into syncytiotrophoblasts, there is a further developmental increase in the capacity of syncytiotrophoblasts to produce CS with advancing stages of baboon pregnancy. Placentas were obtained from baboons in early (days 48-62), mid (days 97-110), and late (days 161-175) gestation (term = 184 days), and CS messenger ribonucleic acid (mRNA) and protein levels were determined in a syncytiotrophoblast-rich cell fraction isolated by Percoll gradient centrifugation. CS mRNA levels in syncytiotrophoblasts, expressed as a ratio of beta-actin, exhibited a progressive increase from early (0.04 +/- 0.04 relative arbitrary units) to mid (2.37 +/- 0.33; P < 0.001) to late (3.66 +/- 0.39; P < 0.05) gestation. Levels of the 22 kDa CS protein were very low on days 48-55 (0.83 +/- 0.09 arbitrary units), increased 10-fold (P < 0.001) on days 57-60 (8.11 +/- 0.68), and increased (P < 0.001) to a maximum of 14.58 +/- 0.58 near term. CS mRNA levels in whole placental villous tissue increased (P < 0.05) between early (0.89 +/- 0.48) and mid (2.97 +/- 0.47) gestation, then remained constant. CS protein exhibited a similar increase (P < 0.001) in villous tissue between early (2.32 +/- 0.40) and mid (6.07 +/- 0.24) gestation, then remained constant. The increase in mRNA and protein levels of CS in the placenta was accompanied by a progressive (P < 0.001) rise in serum CS. We conclude that in addition to the morphological differentiation of cytotrophoblasts into syncytiotrophoblasts that has been well established to result in the onset of CS biosynthesis, villous syncytiotrophoblasts undergo functional/biochemical differentiation thereafter, manifested as an increase in the capacity for the synthesis of CS. PMID- 9398723 TI - Calcium absorptive effects of vitamin D and its major metabolites. AB - The absorptive response to graded doses of vitamin D3, 25(OH)D, and 1,25(OH)2D was measured in healthy adult men after treatment periods of eight, four, and two weeks, respectively. While no relationship was found between baseline absorption and serum vitamin D metabolite levels, all three vitamin D compounds significantly elevated 45Ca absorption from a 300 mg calcium load given as part of a standard test meal. 1,25(OH)2D was active even at the lowest dose (0.5 microgram/day), and the slope was such that doubling of absorption would occur at an oral dose of approximately 3 micrograms/day. 25(OH)D was also active in elevating absorption and did so without raising total 1,25(OH)2D levels. On the basis of the dose response curves for 1,25(OH)2D and 25(OH)D, the two compounds exhibited a molar ratio for physiological potency of approximately 100:1. The absorptive effect of vitamin D3 was seen only at the highest dose level (1250 micrograms, or 50,000 IU/day) and was apparently mediated by conversion to 25(OH)D. Analysis of the pooled 25(OH)D data from both the 25(OH)D- and vitamin D3-treated groups suggests that approximately one eighth of circulating vitamin D like absorptive activity under untreated conditions in winter may reside in 25(OH)D. This is a substantially larger share than has been predicted from studies of in vitro receptor binding. PMID- 9398724 TI - Persistent elevation and metabolic dependence of circulating E-selectin after delivery in women with gestational diabetes mellitus. AB - The increased risk of premature atherosclerosis in noninsulin-dependent diabetes mellitus (NIDDM) might be related in part to augmented expression of endothelial adhesion molecules (AMs). So far it is, however, unknown whether increased circulating (c) AMs in NIDDM are only a consequence of this disease or also involved in its sequelae. To determine the presence of cAMs in a population at increased risk for subsequent development of NIDDM, we analyzed fasting and postprandial [oral glucose tolerance test (OGTT): 100 g] serum concentrations of circulating E-selectin, vascular cell adhesion molecule-1 (cVCAM-1), and intercellular adhesion molecule-1 (cICAM-1) in pregnant women with either gestational diabetes (GDM) or normal glucose tolerance (NT) before and after delivery vs. nonpregnant healthy women (C). During pregnancy cE-selectin and cVCAM-1 were elevated in both GDM and NT vs. nonpregnant females (P < 0.01 vs. C). Following delivery, all GDM females regained normal glucose tolerance according to OGTT criteria, but showed slightly higher postprandial [area under the curve (AUC)180 min] glycemia and HbA1c values than nonpregnant healthy women (P < 0.05), indicating persisting subtle abnormalities in carbohydrate metabolism. cE-selectin and cVCAM-1 remained increased in GDM (P < 0.01 vs.C) after delivery, but fell to normal in NT (P < 0.05 before vs. after delivery). Furthermore, a correlation was seen in GDM females between cE-selectin and HbA1c (P < 0.005), fasting glucose (P < 0.01), and insulin (P < 0.05) as well as postprandial (AUC180 min) glucose and insulin concentrations (P < 0.05) during OGTTs, both before and after delivery. ICAM-1, however, did not differ significantly between groups. In summary, GDM is characterized by persistently raised levels of cE-selectin and cVCAM-1 12 weeks after delivery. Whether these persistent elevations of cE-selectin and cVCAM-1 reflect early vascular injury or represent a risk factor for atherosclerosis in women at increased risk for NIDDM remains to be determined. PMID- 9398726 TI - Codon 17 polymorphism of the cytotoxic T lymphocyte antigen 4 gene in Hashimoto's thyroiditis and Addison's disease. AB - Endocrine autoimmune disorders share susceptibility and resistance factors of the human leukocyte antigen system on the short arm of chromosome 6, but other gene loci also contribute to predisposition and protection. Because the cytotoxic T lymphocyte antigen 4 (CTLA4) alanine-17 encoded by the CTLA4 gene on chromosome 2q33 confers susceptibility to Graves' disease, as well as to type 1 (insulin dependent) diabetes mellitus, we investigated this dimorphism in the other endocrine autoimmune disorders: Hashimoto's thyroiditis and Addison's disease. We analyzed the CTLA4 exon 1 polymorphism (49 A/G) in 73 patients with Hashimoto's thyroiditis, 76 with Addison's disease, and 466 healthy controls. This dimorphism corresponds to an aminoacid exchange (Thr/Ala) in the leader peptide of the expressed protein. CTLA4 alleles were defined by PCR, single-strand conformational polymorphism analysis, and restriction fragment length polymorphism analysis using BbvI. Patients with Hashimoto's thyroiditis had significantly more Ala alleles than controls, both as homozygotes (22% vs. 15%) and heterozygotes (53% vs. 46%), and less Thr than controls as homozygotes (25% vs. 39%), P < 0.04. The phenotypic frequency for Ala was significantly higher in patients (75%), compared with controls (61%), P < 0.03. Patients with Addison's disease did not differ significantly from controls, but those carrying the suceptibility marker, human leukocyte antigen DQA1*0501, were significantly more CTLA4 Ala17 positive than controls with the same DQA1 allele (P < 0.05). In conclusion, an alanine at codon 17 of CTLA4 confers genetic susceptibility to Hashimoto's thyroiditis, whereas this applies only to the subgroup of DQA1*0501+ patients with Addison's disease. PMID- 9398725 TI - Identification, characterization, and biological activity of endothelin receptors in human ovary. AB - We previously reported the expression of endothelin-1 (ET-1) in granulosa cells (GCs) of the human ovary and the presence of ET-1-like immunoreactivity in human follicular fluid obtained from women in an in vitro fertilization program. In follicular fluid, but not in plasma, the levels of ET-1-like immunoreactivity were higher in gonadotropin-stimulated vs. spontaneous cycles, suggesting hormonal regulation of follicular ET-1. To identify and characterize ET receptors in human ovary, we performed autoradiography, radioligand binding, and functional studies. Mathematical analysis of families of self- and cross-competition curves among [125I]ET-1, [125I]ET-3, and selective analogs indicates that human ovary expresses both subtypes of ET receptors, i.e. ETA and ETB receptors. However, the concentration of the ETB site was 100-fold lower than that of the ETA one. By using [125I]ET-1, we demonstrated that the density of binding sites in human ovary is not affected by the hormonal milieu (similar concentrations in normal cycling, postmenopausal, and combined oral contraceptive-treated women). In situ binding studies indicate that the majority of ETA and ETB receptors are expressed in the blood vessels of the ovary. In particular, ETA receptors are abundant in the ovulatory follicles and localized in the theca interna, in close proximity to the granulosa layer. Few GCs of the ovulatory follicle were specifically labeled. Conversely, in the rat ovary, used as a control, ETB receptors were predominantly expressed and localized in GCs. Accordingly, ETB receptors negatively regulated estrogen accumulation in rat GCs. In human granulosa-luteal cells, neither ET-1 (unselective ligand) nor ET-3 or sarafotoxin 6c (ETB ligands) affected estrogen or progesterone secretion. ET-1 was 2.5-fold more potent than noradrenaline in eliciting contraction of ovarian artery, acting through the ETA receptor. Our results indicate that in human ovary, at variance with rat ovary, the endothelin system is primarily involved in the regulation of ovarian blood flow and not steroidogenesis. PMID- 9398727 TI - Contractile properties and fiber type distribution of quadriceps muscles in adults with childhood-onset growth hormone deficiency. AB - Adults with GH deficiency (GHD) report weakness and fatigability. The origin of such symptoms is still debated. This work aimed to clarify whether weakness and fatigability depend on impairment of skeletal muscle contractile capacity. Five males with childhood-onset GHD (age +/- SE, 29.6 +/- 1.9) and 13 age- and sex matched controls were enrolled in the study. Quadriceps muscle cross-sectional area (CSA), strength, twitch characteristics, and fatigue index of voluntary and electrically evoked contractions were determined in vivo in all subjects. Fiber type distribution and CSA of identified types of skeletal fibers were determined on needle biopsy samples of the vastus lateralis muscle of all subjects. Fiber type distribution was assessed on the basis of myosin heavy chain (MHC) isoform composition determined by electrophoresis on polyacrylamide gels. Fiber CSA was determined on cross-cryosections of fiber bundles immunostained by monoclonal antibodies against MHC isoforms. Absolute values of strength and fiber CSA of quadriceps were significantly lower in patients affected by GHD than in controls. However, once strength and fiber CSA were normalized for quadriceps CSA and subject height, respectively, differences disappeared. No difference was found between GHD patients and controls for quadriceps muscle twitch characteristics, fatigue index, and fiber type distribution. The results reported here suggest that weakness and fatigability in childhood-onset GHD do not have a skeletal muscle origin. PMID- 9398728 TI - Isoproterenol and somatostatin decrease plasma leptin in humans: a novel mechanism regulating leptin secretion. AB - In cultured adipocytes, leptin is increased by insulin and decreased by cAMP. In animal models, insulin and agents that increase intracellular cAMP have been shown to similarly affect plasma leptin in vivo. This study was undertaken to test the hypothesis that in humans increased cAMP induced by isoproterenol would decrease leptin. Five groups of normal weight subjects were studied; 1) subjects infused with isoproterenol at a rate of 24 ng/kg/min (ISO24); 2) subjects infused with isoproterenol at a rate of 8 ng/kg/min (ISO8); 3) subjects infused with somatostatin/insulin/GH followed by coinfusion with 8 ng/kg/min isoproterenol (ISO8 + SRIH); 4) subjects infused with somatostatin/insulin/GH alone (SRIH); and 5) control subjects infused with saline (NS). ISO24 infusion resulted in a 27% decrease in plasma leptin over 120 min. ISO24 also increased plasma insulin over the infusion. ISO8 resulted in a 16% decrease in leptin. Saline did not change leptin. SRIH alone decreased leptin 19% over the first 120 min, however no additional fall was seen over the next 120 min the SRIH group. Nonetheless, the addition of 8 ng/kg/min ISO during the second 120 min (ISO8 + SRIH) caused a 15% further decline in plasma leptin. Therefore both isoproterenol and somatostatin reduce plasma leptin in humans. The effect of isoproterenol is likely mediated by beta-adrenergic receptors, whereas the effect of somatostatin suggests a novel mechanism for the regulation of leptin. PMID- 9398729 TI - Expression of functional leptin receptors in the human ovary. AB - The size of body fat stores is known to influence fertility, indicating a link between adipose tissue and the reproductive system. Studies in mice have identified the adipocyte-derived hormone, leptin (Ob protein), as a possible mediator of this effect. The aim of this study was to investigate the possibility that leptin may have direct effects on the human ovary. To probe this hypothesis we first analyzed the expression of leptin receptors in the human ovary. Transcripts encoding both the long and short isoforms of the leptin receptor were present in human granulosa cells and thecal cells; however, the short isoforms were expressed at much higher levels. Immunoreactive leptin was present in follicular fluid at levels similar to those found in serum. ob gene expression, however, was undetectable in the ovary, as determined by reverse transcription PCR, whereas it was easily detected in adipose tissue. To determine whether leptin could induce a biological response in ovarian cells, we examined the effect of leptin on estradiol production in cultured granulosa cells. Leptin (100 ng/mL) inhibited LH (0.1 ng/mL)-stimulated estradiol production. In contrast, leptin had no effect on estradiol production in the absence of LH. In conclusion, this study has demonstrated that the leptin receptor is expressed in the human ovary, that leptin is present in follicular fluid, and that leptin can induce a biological response in ovarian cells. These results suggest that leptin may have a direct effect on the human ovary. PMID- 9398730 TI - Serum leptin and energy expenditure in children. AB - Leptin has been hypothesized to play an important role in energy balance by affecting both energy intake and energy expenditure. The purpose of our study was to determine the relationship between fasting serum leptin concentrations and measures of energy expenditure in prepubertal children. We measured total energy expenditure (TEE; by the doubly labeled water technique), resting energy expenditure (REE; after an overnight fast), activity energy expenditure (AEE; TEE REE), body composition (by dual energy x-ray absorptiometry), and fasting serum leptin concentration (by RIA) in 76 children. Simple correlations showed that all measures of energy expenditure (TEE, REE, and AEE) were positively related to the serum leptin concentration (r = 0.50, P < 0.001; r = 0.45, P < 0.001; and r = 0.30, P < 0.01, respectively). However, after adjusting for body composition (fat free mass and fat mass), gender, and ethnicity, serum leptin concentrations were not related to any measure of energy expenditure (TEE, P = 0.61; REE, P = 0.97; AEE, P = 0.65). These latter findings were further confirmed using structural equation models with leptin and energy expenditure as dependent variables, and fat-free mass and fat mass as independent variables. Results from these models showed no direct effect of leptin and no indirect effect of fat mass (through leptin) on any measure of energy expenditure, when a path between fat mass and energy expenditure was present in the model. Thus, our data do not support the hypothesis that the serum leptin concentration (independent of fat mass) is related to measures of energy expenditure in children. PMID- 9398731 TI - Evidence for production and functional activity of nitric oxide in seminiferous tubules and blood vessels of the human testis. AB - Previous studies have demonstrated that nitric oxide (NO) influences Leydig cell function. Here we provide evidence for NO production and activity in seminiferous tubules and blood vessels of the human testis. By immunohistochemistry, the soluble guanylyl cyclase (sGC), the intracellular NO receptor, and the second messenger, cyclic guanosine monophosphate (cGMP), were detected in myofibroblasts of the peritubular lamina propria in Sertoli cells, as well as in endothelial and smooth muscle cells of testicular blood vessels. Performed with isolated tubules and blood vessels, the biological activity of sGC could be proved by cGMP generation in response to treatments with the NO donor, sodium nitroprusside. The endothelial and neuronal subtypes of NO synthase (NOS) were localized immunohistochemically to the same cell types that express sGC and cGMP. In isolated tubules and vessels, the presence of endothelial NOS and neuronal NOS was confirmed by immunoblotting, and NOS activity was demonstrated by decreased cGMP production upon incubation with the NOS inhibitor L-nitro arginine methylester. These findings show that peritubular cells, Sertoli cells, and testicular blood vessels may be sites of NO production and activity, possibly involved in relaxation of seminiferous tubules and blood vessels to modulate sperm transport and testicular blood flow, respectively. PMID- 9398732 TI - Comparison of hormone-sensitive lipase activity in visceral and subcutaneous human adipose tissue. AB - The possible role of hormone-sensitive lipase (HSL) in determining regional differences in lipolysis activation in humans was studied in vitro. Small adipose tissue biopsies were obtained from the abdominal sc and omental regions during surgery in 21 subjects spanning a wide range of body mass index (22-50 kg/m2). In lipolysis experiments, isolated fat cells were incubated with lipolytic agents acting at different levels in the lipolytic cascade. The activity and messenger ribonucleic acid expression of HSL were determined. The maximum lipolytic capacity was higher in sc than in omental fat cells as were HSL activity and messenger ribonucleic acid expression. The maximum lipolysis rate was significantly correlated to HSL activity. This is in accordance with the role of HSL as the rate-limiting step of lipolysis. However, adipocytes were 24% larger in the sc than in the omental region, and the lipolysis rate was significantly correlated to fat cell size regardless of either the region of origin or gender. This indicates that the regulation of HSL activity in healthy subjects, which appears to occur at a transcriptional level, is to a large extent dependent on fat cell size. PMID- 9398733 TI - Dose-dependent effects of recombinant human interleukin-6 on glucose regulation. AB - Inflammatory cytokines have metabolic actions that probably contribute to the general adaptation of the organism during infectious or inflammatory stress. To examine the effects of interleukin 6 (IL-6), the main circulating cytokine, on glucose metabolism in man, we performed dose-response studies of recombinant human IL-6 in normal volunteers. Increasing single doses of IL-6 (0.1, 0.3, 1.0, 3.0, and 10.0 mg/Kg BW) were injected sc in 15 healthy male volunteers (3 in each dose) after a 12-h fast. All IL-6 doses were tolerated well and produced no significant adverse effects. We measured the circulating levels of glucose, insulin, C-peptide, and glucagon at baseline and half-hourly over 4 h after the IL-6 injection. Mean peak plasma levels of IL-6 were achieved between 120 and 240 min and were 8, 22, 65, 290, and 4050 pg/mL, respectively, for the 5 doses. After administration of the 2 smaller IL-6 doses, we observed no significant changes in plasma glucose levels, which, because of continued fasting, decreased slightly over time. By 60 min after the 3 higher IL-6 doses, however, the decline in fasting blood glucose was arrested, and glucose levels increased in a dose dependent fashion. The concurrent levels of plasma insulin and C-peptide were not affected by any IL-6 dose. In contrast, IL-6 caused significant increases in plasma glucagon levels, which peaked between 120 and 150 min after the IL-6 injection. In conclusion, sc IL-6 administration induced dose-dependent increases in fasting blood glucose, probably by stimulating glucagon release and other counteregulatory hormones and/or by inducing peripheral resistance to insulin action. PMID- 9398734 TI - Nitric oxide inhibits corticotropin-releasing hormone exocytosis but not synthesis by cultured human trophoblasts. AB - Nitric oxide (NO) plays an important role in many cell-cell signaling systems, but its mechanism of action is variable. We have previously reported that NO reduces secretion of the peptide hormone, CRH, from cultured placental cells and the perfused placenta. Because placental CRH production seems linked to human parturition, we wished to explore the mechanism of action of NO in this setting in more detail. We report here that in the placenta, NO specifically inhibited CRH exocytosis, not synthesis, and that endogenous NO affects this process. Cytotrophoblasts were prepared from term human placentas and cultured as monolayers. CRH immunoreactivity in the cell supernatants and cell extracts were measured by RIA. CRH messenger RNA was determined by Northern blot analysis. Sodium nitroprusside (SNP; 1-100 mumol/L) and S-nitroso-N-acetyl-penicillamine (SNAP; 1-100 mumol/L), NO donors, significantly reduced basal CRH concentration in the media, while increasing the concentration of CRH in the cells (P < 0.01), suggesting that exocytosis of CRH was inhibited. These effects could be attenuated by the NO scavenger hemoglobin (20 micrograms/mL). KCl (45 mmol/L), which causes exocytosis by depolarizing the cell membrane, increased CRH release by 2- to 3-fold, and this was inhibited by SNP. Basal release of CRH was augmented by the NO synthase competitive inhibitor N omega-L-arginine methyl ester (1 mmol/L; P < 0.01) and the guanylate cyclase inhibitor, LY83583 (1 mumol/L; P < 0.01). The inhibitory effect of SNP was also blocked by LY83583. CRH messenger RNA content did not change when the placental cells were incubated with SNP, N omega-L-arginine methyl ester, and LY83583 for 6 and 24 h, and this was consistent with studies showing that total CRH immunoreactivity (cells plus media) did not change in the presence of SNP. These studies indicate that exogenous NO inhibits CRH exocytosis, rather than biosynthesis, by human trophoblasts and that endogenous NO has tonic inhibitory effects on CRH release by these cells. The inhibitory effect of NO on basal and stimulated CRH release by placental trophoblasts seems to be a guanylate cyclase-mediated inhibition of exocytosis. PMID- 9398735 TI - A novel point mutation in the intracellular domain of the ret protooncogene in a family with medullary thyroid carcinoma. AB - Specific mutations in the ret protooncogene have been found associated with multiple endocrine neoplasia type 2A (MEN 2A) and type 2B (MEN 2B) and familial medullary thyroid carcinoma (FMTC). Mutations in one of five cysteine residues in the extracellular domain have been found in over 95% of families with MEN 2A and 88% of families with FMTC. In MEN 2B patients, a specific mutation at codon 918, substituting a threonine for a methionine, has been found in 95% of cases. In FMTC, in addition to the mutations of the extracellular cysteines, three intracellular base pair changes have been reported at codons 768 and 804. Here we describe a novel intracellular mutation in exon 15 of the ret gene that leads to the substitution of an alanine for a serine at codon 891 in a family with medullary thyroid carcinoma. This amino acid change may be important in determining substrate specificity or, alternatively, may play a role in ATP binding. PMID- 9398736 TI - Resistance of gonadotropin releasing hormone drive to sex steroid-induced suppression in hyperandrogenic anovulation. AB - Women with hyperandrogenic anovulation (HAA) exhibit increased GnRH drive, as evidenced by a faster LH pulse frequency that slows in response to progestin induced opioidergic tone. To determine whether increased GnRH-LH drive in HAA reflects altered sex steroid exposure caused by chronic anovulation or is an intrinsic hypothalamic attribute, we compared the pulsatile LH response to oral contraceptive (OC)-induced suppression in seven women with HAA, with that of seven eumenorrheic women (EW). LH levels were determined at 10-min intervals for 12 h after 19-21 days of OC use and 5-7 days after cessation. Testosterone, androstenedione, estradiol, FSH, and LH levels were determined at weekly intervals before, during, and after OC use. LH pulse number/12 h was higher (P < 0.001) in HAA during and after OCs, when compared with that of EW. Mean LH was increased in HAA before, during, and after OCs. Testosterone, androstenedione, and estradiol levels were higher in HAA before OCs, but they decreased to similar levels during OC use in both groups. FSH concentrations were similar before and during OCs but rose more after cessation of OCs in EW. These findings indicate that GnRH drive in HAA is resistant to OC-induced suppression and, therefore, could be an intrinsic hypothalamic attribute. PMID- 9398737 TI - Pituitary adenomas: screening for G alpha q mutations. AB - Mutant, guanosine triphosphatase-deficient, alpha-subunits of the G protein, Gs, gsp ocogene have been discovered in 40% of GH-secreting pituitary adenomas. Therefore, we hypothesized that a novel G protein class, G alpha q, involved in pituitary signal transduction, might be involved in pituitary tumorigenesis. Recombinant mutations of G alpha q result in constitutive activation of phospholipase C and have transforming activity. Therefore, we screened tumor samples from 37 pituitary adenomas for the presence of activating mutations of the G alpha q gene. Importantly, our sample contains 8 FSH and LH adenomas. In the pituitary gland, FSH and LH are linked to the GnRH-G alpha q signaling cascade, making these tumors a logical choice for screening for G alpha q mutations. Complementary DNA (cDNA) was synthesized by RT-PCR with G alpha q specific primers to exclude pseudogene transcripts. Fragments of G alpha q cDNA encompassing residues (Arg183, Gln209) were screened by single-strand conformation polymorphism and then sequenced in both directions. No mutations were detected. We conclude that mutations in these regions of the G alpha q cDNA occur infrequently, if at all, in human pituitary adenomas. Alternative mechanisms underlying pituitary tumorigenesis should be explored. PMID- 9398738 TI - Endometrial alpha-2 macroglobulin; localization by in situ hybridization and effect on mouse embryo development in vitro. AB - alpha-2 macroglobulin (A2M) is a 718,000-kDA broad spectrum plasma protease inhibitor whose production by the human endometrium was recently reported. The multifunctional A2M receptor, also known as low-density lipoprotein receptor related protein, was also recently immunolocalized to the endometrial stroma. The objective of this study was to further characterize the endometrial site of expression of A2M, and to study its effects on mouse embryo development in vitro, to gain some insight into the functional significance of its endometrial production. Formalin-fixed, paraffin-embedded human endometrium from hysterectomy and endometrial biopsy specimen was used for in situ hybridization analysis, with 35S-labeled riboprobes representing subcloned A2M complementary DNA (cDNA) fragments. Duplicate sections of human endometrium were hybridized with sense and antisense probe and coated with photographic emulsion. Resultant autoradiograms were analyzed qualitatively by light- and darkfield microscopy and quantitatively by a computerized analysis of the signal intensity. Immunohistochemistry and immunoblotting for endometrial tissues were performed using an affinity-purified polyclonal antibody to human A2M. The effect of A2M on mouse embryo development was studied by exposure of one cell mouse embryo in culture to physiological concentrations of biologically active and inactive A2M. Expression signals for A2M were more numerous and intense in the secretory endometrium, compared with proliferative endometrium. Endothelial cells lining the endometrial blood vessels seemed to be the main source of A2M expression. The A2M expression signals in secretory endothelium were 2- to 3-fold stronger than the proliferative endothelium, suggesting transcriptional activation of A2M expression in the secretory endothelium. Glandular expression was observed in secretory endometrium from two patients with endometriosis. Ectopic endometrial tissues also produced A2M. A2M at concentrations of 400-500 mumol/L significantly inhibited blastocyst development of mouse embryos in vitro. A2M is expressed predominantly by the endometrial endothelial cells and may be involved in endometrial physiology. Physiological concentrations of A2M inhibit mouse embryo development in vitro, suggesting that endometrial production of A2M may play a role in regulating preimplantation embryo development. PMID- 9398739 TI - Subcutaneous adipose tissue releases interleukin-6, but not tumor necrosis factor alpha, in vivo. AB - We measured arterio-venous differences in concentrations of tumor necrosis factor alpha (TNF alpha) and interleukin-6 (IL-6) across a sc adipose tissue bed in the postabsorptive state in 39 subjects [22 women and 17 men; median age, 36 yr (interquartile range, 26-48 yr); body mass index, 31.8 kg/m2 (range, 22.3- 38.7 kg/m2); percent body fat, 28.7% (range, 17.6-50.7%)]. A subgroup of 8 subjects had arteriovenous differences measured across forearm muscle. Thirty subjects were studied from late morning to early evening; 19 ate a high carbohydrate meal around 1300 h, and 11 continued to fast. We found a greater than 2-fold increase in IL-6 concentrations across the adipose tissue bed [arterial, 2.27 pg/mL (range, 1.42-3.53 pg/mL); venous, 6.71 pg/mL (range, 3.36-9.62 pg/mL); P < 0.001], but not across forearm muscle. Arterial plasma concentrations of IL-6 correlated significantly with body mass index (Spearman's r = 0.48; P < 0.01) and percent body fat (Spearman's r = 0.49; P < 0.01). Subcutaneous adipose tissue IL 6 production increased by the early evening (1800-1900 h) in both subjects who had extended their fasting and those who had eaten. Neither deep forearm nor sc adipose tissue consistently released TNF alpha [across adipose tissue: arterial, 1.83 pg/mL (range, 1.36-2.34 pg/mL); venous, 1.85 pg/mL (range, 1.44-2.53 pg/mL); P = NS: across forearm muscle: arterial, 1.22 pg/mL (range, 0.74-2.76 pg/mL); venous, 0.99 pg/mL (range, 0.69-1.70 pg/mL); P = NS]. Although both IL-6 and TNF alpha are expressed by adipose tissue, our results show that there are important differences in their systemic release. TNF alpha is not released by this sc depot. In contrast, IL-6 is released from the depot and is thereby able to signal systemically. PMID- 9398740 TI - Expression of variant forms of insulin receptor substrate-1 identified in patients with noninsulin-dependent diabetes mellitus. AB - Several polymorphisms have been identified in the amino acid sequence of human insulin receptor substrate-1 (IRS-1). Some of the variant sequences have been reported to be increased in prevalence among patients with noninsulin-dependent diabetes mellitus (NIDDM). This observation led to the hypothesis that these amino acid substitutions may impair the function of IRS-1, thereby causing the insulin resistance seen in patients with NIDDM. To address this question, we have designed studies to evaluate the effects of three variant sequences identified in our laboratory: Gly819-->Arg, Gly972-->Arg, and Arg1221-->Cys. We constructed four IRS-1 expression vectors for transfection in COS-7 cells: wild-type, single mutant (Gly819-->Arg), double mutant (Gly819-->Arg; Gly972-->Arg), and triple mutant (Gly819-->Arg; Gly972-->Arg; Arg1221-->Cys) IRS-1. The mutations did not alter the level of expression or the extent of insulin receptor-mediated tyrosine phosphorylation of recombinant IRS-1. Moreover, the mutations did not lead to a detectable impairment in the association of recombinant IRS-1 with important downstream effectors, including the p85 subunit of phosphatidylinositol 3-kinase and growth factor receptor-binding protein-2. We conclude that these amino acid substitutions do not appear to cause a major defect in the function of IRS-1, as judged by our assays. However, this type of assay probably lacks the sensitivity to detect subtle functional defects. In light of the suggestive associations observed in epidemiological studies, it is premature to totally discard the hypothesis that variant sequences of IRS-1 may contribute to the pathogenesis of NIDDM. Nevertheless, our studies cannot be interpreted as lending support to that hypothesis. PMID- 9398741 TI - Stimulation of mitochondrial fatty acid oxidation by growth hormone in human fibroblasts. AB - In vivo administration of GH induces lipolysis and lipid oxidation. However, it is not clear whether the stimulation of lipid oxidation is a direct effect of GH or is driven by increased substrate supply secondary to lipolysis. An in vitro bioassay has been established for assessing beta-oxidation of fatty acids in mitochondria, based on the measurement of conversion of tritiated palmitic acid to 5H2O by fibroblasts in culture. We have modified this assay to investigate whether GH stimulates fatty acid oxidation. GH stimulated oxidation of palmitic acid maximally by 26.7 +/- 2.5% (mean +/- SEM; P < 0.0001). The stimulation was biphasic, with the oxidation rate increasing with increasing GH concentration to a peak response at 1.5 nmol/L and declining to a level not significantly different from control thereafter. Insulin-like growth factor-I at concentrations of up to 250 nmol/L had no significant effect on fatty acid oxidation. GH-binding protein attenuated the effect of GH. An anti-GH receptor (GHR) antibody (MAb263), which dimerizes the receptor and induces GH-like biological actions, significantly stimulated fatty acid oxidation. Another anti-GHR antibody (MAb5), which prevents receptor dimerization, suppressed GH action. In summary, GH directly stimulated fatty acid oxidation, an action not mediated by insulin-like growth factor-I. Dimerization of GHRs was necessary for this effect. This bioassay is a practical tool for studying the regulatory effects of GH on lipid oxidation. PMID- 9398742 TI - Telomerase activity in human thyroid carcinomas originating from the follicular cells. AB - Telomerase activity is known to be absent from most normal and well differentiated tissues, although being detectable in the vast majority of malignant tumors. An increasing number of reports demonstrate that telomerase may be activated in benign tumors, such as adenomas. We have investigated a series of normal and neoplastic thyroid tissues for the presence of telomerase activity. As expected, all normal thyroid tissues (n = 20) had no display of telomerase activity. Amongst cancers, the incidence of telomerase activity varied with the histological subtypes. Telomerase activity was present in only 3/15 cases (20%) of papillary carcinomas. Telomerase activity was more frequently detected in follicular (4/6) and in undifferentiated (2/3) carcinomas. Unexpectedly, one case (1/12) of adenoma contained high levels of telomerase activity. Taken together, these results indicate that telomerase may play some role in the pathogenesis of thyroid tumors, in particular in follicular and undifferentiated carcinomas that are known to have the most aggressive behavior. PMID- 9398743 TI - Thyroid hormone autoantibodies elicited by diagnostic fine needle biopsy. AB - Based on the knowledge that diagnostic fine needle biopsy of the thyroid (FNAB) results in a prompt increase in circulating thyroglobulin (Tg); we evaluated whether Tg is indeed the postulated antigen for circulating antibodies against thyroid hormones (THAb). Preliminarily, we verified that FNAB causes the release into the bloodstream of iodinated, heterologous, and thus potentially immunogenic, molecules of Tg. Of the initially enrolled 400 patients, 214 had a number of blood drawings sufficient to evaluate over time (before FNAB and 1-3 h, 3 days, 15 days, 30 days, 3 months, 6 months, and 12 months after FNAB) the following parameters: THAb of both IgM and IgG classes, Tg antibodies (TgAb; by a sensitive immunoradiometric assay), and Tg (in the 156 patients who were TgAb negative). We found the following. 1) Serum Tg most often peaks 1-3 h after FNAB (61 +/- 45% of the baseline level; mean +/- SD). 2) Only 7% of the initially TgAb negative patients converted to positive, and only 12% of those initially positive had an increase in the levels of TgAb. 3) THAb were detected in 0 of 400 patients before FNAB, but were found in 9 of 214 (4.2%) after FNAB. This proportion is 2 orders of magnitude higher than that (149 of 369,000 or 0.04%) found in consecutive patients attending European thyroid clinics. Of the 9 cases, 6 had Hashimoto's thyroiditis (HT), 2 had euthyroid colloid goiter, and 1 had Hurthle cell carcinoma. In the 5 of 9 cases who were TgAb negative, the post-FNAB increment in Tg was 21-99%, i.e. lower than that of the majority of patients (101 12,500%). 4) THAb were of the IgM class in all 9 (6 against T3 and 3 against T4), and were accompanied and/or followed up to 3 months after FNAB by IgG-THAb of the same specificity (2 against T3 and 1 against T4) in 3 cases. In a fourth case, IgM-T3 were followed by a long-lasting synthesis of IgG-T3 (i.e. up to 1 yr post FNAB). All 4 cases with IgG-THAb had HT and remained TgAb positive. 5) In the 2 HT and the 3 non-HT patients with undetectable TgAb, THAb were of the IgM class only. 6) In the HT group, 2 risk factors for the development of post-FNAB THAb appeared to be pre-FNAB TgAb levels below 400 U/mL that did not increase after FNAB and Tg released from a colloid nodule. We conclude that Tg release from the thyroid is sufficient to elicit THAb synthesis. In patients with autoimmune thyroid disease (HT), this synthesis occurs with a frequency 10-fold higher than that in patients with nonautoimmune thyroid diseases (21% vs. 2%). However, in only a fraction of patients with autoimmune disease, who need to be TgAb positive by a sensitive assay, the primary immune response (IgM) is followed by a secondary one (IgG). As, once present, this secondary response is long lasting in only a minority of our patients, we think that this could contribute to the rarity of naturally occurring THAb. PMID- 9398744 TI - A novel transcript for the thyrotropin-releasing hormone receptor in human pituitary and pituitary tumors. AB - We measured the amounts of TRH receptor (TRHR) messenger ribonucleic acid (mRNA) in human normal pituitary and pituitary tumors and found a novel transcript of the TRHR gene. Competitive PCR revealed expression of the TRHR mRNA in all pituitary adenomas examined, and its level was variable and similar to that in the normal pituitary. When the C-terminal region was amplified by PCR, an additional short product was observed. Cloning and sequence analysis of this short fragment revealed that the deleted sequence corresponded exactly to the 5' sequence of exon 3, indicating alternative splicing of the TRHR mRNA. This alternative splicing resulted in a frame shift, yielding a C-terminal truncated protein (HTRHR2) on translation. Expression analysis of HTRHR2 in Chinese hamster ovary cells showed no significant binding to [3HIMeTRH or response of intracellular calcium to TRH administration. However, the mRNA ratio of HTRHR2 vs. the wild type (HTRHR1) was significantly different among pituitary tumors. The highest ratio was observed in prolactinomas (30%), and almost no detectable expression was found in GH-producing tumors. These findings indicate that this novel transcript of the human TRH receptor gene is produced in a tumor-specific manner and may be a useful parameter for evaluation of individual pituitary tumors. PMID- 9398745 TI - Identification of constitutively activating somatic thyrotropin receptor mutations in a subset of toxic multinodular goiters. AB - Constitutively activating mutations in the TSH receptor (TSHR) gene and in the Gs alpha gene are frequent molecular causes for solitary toxic nodules of the thyroid. However, the etiology of toxic multinodular goiter is still largely unknown. Therefore, DNA from nodular and quiescent surrounding tissue of six patients with toxic multinodular goiters was screened for mutations in exons 9 and 10 of the TSHR gene and exons 7-10 of the Gs alpha gene by direct automated sequencing. In one patient, two different somatic TSHR mutations were identified in two different toxic nodules (L632I and F631L). In another patient, two different toxic nodules harbored the same TSHR mutation (I630L), whereas only one TSHR mutation (F631L) was identified in one of the two toxic nodules of an additional patient. In the other three patients, no mutations could be found in exons 9 and 10 of the TSHR gene or in exons 7-10 of the Gs alpha gene. Our results demonstrate that not only solitary toxic adenomas but also toxic multinodular goiters can be caused by constitutively activating mutations of the TSHR. In addition to mutations in the TSHR and possibly in Gs alpha, there are probably other still unknown mechanisms that cause hot nodules in toxic multinodular goiters. PMID- 9398746 TI - Identification of a new thyrotropin receptor germline mutation (Leu629Phe) in a family with neonatal onset of autosomal dominant nonautoimmune hyperthyroidism. AB - Constitutively activating germline mutations in the TSH receptor (TSHR) gene have been identified as a cause of autosomal dominant nonautoimmune hyperthyroidism and sporadic congenital hyperthyroidism. We report a 10-yr-old boy and his 31-yr old mother, both presenting with a history of recurring toxic thyroid hyperplasia and no evidence for autoimmune thyroid disease. In the boy, onset of hyperthyroidism and goiter was neonatal. In the mother, onset of thyroid disease dates back to early childhood. There was no history of thyroid disease in the rest of the family. Screening for germline mutations in exon 10 of the TSHR was performed by direct sequencing of genomic DNA extracted from peripheral blood leukocytes of both patients. In the boy and his mother, an identical heterozygous TSHR mutation was identified, exchanging leucine for phenylalanine at residue 629 of the TSHR (TTG-->TTT). Transient expression of the mutated TSHR construct in COS-7 cells confirmed the constitutive activity of the new TSHR germline mutation. This is the second family displaying congenital manifestation of hyperthyroidism in familial nonautoimmune hyperthyroidism. PMID- 9398747 TI - Prenatal diagnosis and treatment of dyshormonogenetic fetal goiter due to defective thyroglobulin synthesis. PMID- 9398748 TI - Localization of the steroidogenic acute regulatory protein in human tissues. AB - The rate-limiting step in steroid hormone production in the adrenal cortex and gonads, the translocation of cholesterol from the outer to the inner mitochondrial membranes, is mediated by the steroidogenic acute regulatory protein (StAR). Heretofore, the localization of StAR in human adult and fetal tissues has not been defined. To this end, expression of StAR was detected in formalin-fixed, paraffin-embedded specimens using a polyclonal antiserum raised against recombinant human StAR. Primordial follicles of adult ovaries did not contain StAR, whereas antral follicles stained intensely in the thecal layer, with occasional staining of granulosa cells. Corpora lutea were intensely stained, but with a patchy distribution. Corpora albicantia did not stain. A luteoma of pregnancy stained with patches of moderate intensity. Ovaries with hyperthecosis contained areas of intense thecal staining. An ovarian Leydig cell tumor stained intensely, whereas granulosa cell tumors were negative. Ovarian adenocarcinomas, borderline tumors, teratomas, cystadenomas, and a Brenner tumor displayed no specific StAR immunostaining. Testicular Leydig cells stained moderately to intensely, as did a testicular Leydig cell tumor. Sertoli cells stained weakly in some specimens. Seminomas and testicular germ cell tumors were negative. There was minimal to moderate staining in the adrenal glomerulosa and faciculata and minimal staining in the reticularis, while the medulla was negative. Adrenal cortical adenomas, hyperplasias, and carcinomas all contained areas of StAR staining. The renal distal tubules stained with moderate to marked intensity. Renal carcinomas had occasional modest staining. No immunostaining was found in the placenta. Fetal ovaries contained sporadic stromal cells displaying intense StAR staining, particularly in the hilar region. Oocytes from a 32-week fetal ovary showed moderate to intense staining. Fetal testes displayed intense Leydig cell staining. The neocortex of the fetal adrenal glands displayed only minimal StAR staining, whereas moderate to intense staining was found in the fetal zone. The fetal kidneys had moderate StAR staining of the distal convoluted tubules. We conclude that StAR is localized to normal and neoplastic cells in the gonads and adrenal cortex, which produce large amounts of pregnenolone. StAR protein was not detected in the placenta, documenting that placental progestin synthesis occurs through StAR-independent mechanisms. The presence of StAR in cells that do not express cholesterol side-chain cleavage enzyme cytochrome P450, including renal distal tubules, Sertoli cells, and fetal oocytes, suggests that StAR has roles in metabolic processes in addition to stimulating pregnenolone synthesis. PMID- 9398749 TI - 11 beta-Hydroxysteroid dehydrogenase type II in the human endometrium: localization and activity during the menstrual cycle. AB - The 11 beta-hydroxysteroid dehydrogenase type II enzyme (11 beta HSD2) is a potent inactivator of glucocorticoids and is present in high amounts in the placental syncytiotrophoblast and sodium-transporting epithelia. Placental 11 beta HSD2 is thought to protect the fetus from high circulating levels of maternal glucocorticoids, whereas the renal enzyme is important in conferring aldosterone specificity on the mineralocorticoid receptor. An isoform of 11 beta HSD (11 beta HSD1) is also present in a wide range of tissues, but usually acts as an oxoreductase, converting the biologically inactive cortisone to cortisol. In the present study we have used an immunopurified antibody to the carboxy terminus of human 11 beta HSD2 (HUH23) to demonstrate localization of the enzyme in luminal and glandular epithelia of human endometrium. In some specimens staining was uniformly distributed, but in others there was clear evidence of heterogeneity both between and within epithelia. Although 11 beta HSD2 was found mainly in the cytoplasm, some cells showed evidence of nuclear staining only. Western blot analysis showed a band at 41 kDa in endometrium and myometrium, confirming the presence of 11 beta HSD2. Measurement of activity throughout the menstrual cycle showed that mean levels (+/- SEM) of activity were 156 +/- 17 and 6.1 +/- 1.1 pmol product/min.g homogenate protein for 11 beta HSD2 and 11 beta HSD1, respectively. Patients taking combined estrogen/progesterone contraceptives had significantly lower activities of both enzymes (76 +/- 19 and 1.9 +/- 0.4; both P < 0.01) compared with the control group. 11 beta HSD2 activity was significantly higher in the secretory than in the proliferative phase of the cycle in controls (193 +/- 22 vs. 120 +/- 23; P < 0.05). All groups contained outliers with elevated enzyme activities, with some patients displaying 11 beta HSD2 levels comparable to those observed in human kidney (> 1000 pmol/min.g). Further analysis showed that there was a statistically significant correlation (r = 0.43; P < 0.001) between the levels of 11 beta HSD1 and 11 beta HSD2. There was no detectable mineralocorticoid receptor binding in endometrial cytosols prepared from patients with a range of 11 beta HSD2 activities. It remains to be determined whether elevated or suppressed levels of either isoform are associated with fertility or endometrial pathology. PMID- 9398750 TI - Human estrogen receptor beta-gene structure, chromosomal localization, and expression pattern. AB - The estrogen receptor (ER) is a ligand-activated transcription factor that mediates the effects of the steroid hormone 17 beta-estradiol, in both males and females. Since the isolation and cloning of ER, the consensus has been that only one such receptor exists. The finding of a second subtype of ER (ER beta) has caused considerable excitement amongst endocrinologists. In this article, we present data regarding the genomic structure and chromosomal localization of the human ER beta gene, demonstrating that two independent ER genes do exist in the human. Furthermore, we present data regarding the tissue distribution of human ER beta, showing that this receptor is expressed in multiple tissues. For instance, ER beta is found in developing spermatids of the testis, a finding of potential relevance for the ongoing debate on the effects of environmental estrogens on sperm counts. In addition, we find ER beta in ovarian granulosa cells, indicating that estrogens also participate in the regulation of follicular growth in the human. PMID- 9398751 TI - Genetic control of anti-thyrotropin receptor antibody generation in H-2K mice immunized with thyrotropin receptor-transfected fibroblasts. PMID- 9398752 TI - Presence of leptin in colostrum and/or breast milk from lactating mothers: a potential role in the regulation of neonatal food intake. AB - In neonates both nutrients and regulatory factors are transferred from the mother to the suckling infant via milk. In the present work, it has been shown that human milk contains immunoreactive leptin which is identical to intact human leptin by criteria of charge, size, immunorecognition and SDS-PAGE mobility. In experimental animals it was demonstrated that leptin is transferred from the circulation to mothers' milk, then to the infant's stomach and afterwards to infant blood. Maternal leptin in milk may play a regulatory role in the suckling infant. PMID- 9398753 TI - Familial nonmedullary thyroid carcinomas: a heterogeneous syndrome with different natural history and variable long-term prognosis. PMID- 9398754 TI - 21-Hydroxylase heterozygotism and immune regulation. PMID- 9398755 TI - Potassium and aldosterone secretion in glucocorticoid-remediable aldosteronism. PMID- 9398757 TI - Commentary on article by Rudavsky and Freeman. PMID- 9398759 TI - Leptin levels are elevated despite low thyroid hormone levels in the "euthyroid sick" syndrome. PMID- 9398760 TI - Health care reform and the emergency physician: a personal strategy. AB - Annals' 25th anniversary is a cause for celebration, but the future imposes a new set of ethical questions and moral dilemmas for emergency physicians. Each physician will need to make personal choices consistent with the courage, judgment, integrity, and dedication of past leaders when dealing with the moral dilemmas of the future. PMID- 9398761 TI - The troubled road to universal health care. AB - In our country the increasing commercialization of medicine is taking control of our medical school faculties, hospitals, and education. There is an overemphasis on health care efficiency, with a dramatic decrease in the commitment to research, an increase in the cost of medical education and resultant staggering student debt, an increasing number of medically uninsured, and an ever-widening gap between the best that American medicine can offer and that which the indigent receive. PMID- 9398762 TI - Thoughts on the clinician's response to health care reform: a cautionary note. AB - In the course of contemporary health care discussion, we frequently refer to "health care reform" and its effect on health care delivery. The context within which we use the expression somehow manages to convey the idea that there is some kind of discreet "fait accompli" to which we can point. This basic premise is incorrect and ascribing to it renders much of the discussion about it in error. PMID- 9398763 TI - Health care reform is dead--long live health care reform. AB - The 1993 Clinton health care reform effort was not the end of reform but the inauspicious start of a fiercely contested round of reform that may take another decade or two to complete. The 1993 Clinton plan was just the latest stage of a battle for national action on health care than began with Teddy Roosevelt's promise of compulsory health insurance in the 1912 presidential campaign. PMID- 9398764 TI - Medicine at the millennium. AB - What will the future bring? Will biomedical advances move rapidly and affordably into clinical practice, stimulated by the demands of the educated consumer? Or will changes in the way health care is financed and physician inability to stay abreast of clinical changes widen the gap between high-quality and mediocre medical care? Is there a limit to what we can afford to provide, regardless of availability? PMID- 9398765 TI - The future of emergency medicine. AB - When considering the future of emergency medicine, we need to study the multiple environments that affect the specialty. This review considers three broad environments: federal government, state jurisdictions, and the private system. PMID- 9398766 TI - The future of the private practice of emergency medicine. AB - No specialty better personifies the changes occurring throughout the health care delivery system than emergency medicine. It was just a short 25 years ago that the specialty of emergency medicine, as it is known today, emerged. The full-time staffing of EDs arose out of a need by patients who were presenting in ever increasing numbers to hospital EDs that were staffed by a nurse or part-time physician. The specialty has continued to be responsive to the changing health care system throughout the past 25 years with innovations such as fast track units, observation units, and chest pain treatment center. To develop a vision for the future of the specialty, it is important to first evaluate the current trends in health care and their influences on the specialty. Three significant areas stand out in the current health care landscape: consolidation of hospital systems, emergence of publicly traded physician practice management companies, and the increasing penetration of managed care. PMID- 9398767 TI - Emergency medicine: a plan for the twenty-first century. AB - Emergency physicians, especially those currently charged with fiscal responsibility within their group, know very well the complicated issues of today's health care environment. We must remain open-minded about all the possible relationships that might ensure optimum patient care and our specialty's future. PMID- 9398768 TI - Economic credentialing. AB - In the 1990s, hospital management and trustees introduced the concept of evaluating physicians for appointment, reappointment, and privilege delineation with the addition of financial criteria. Although emergency physicians are advocates for cost-effective care, they must make certain that credentials are determined by the provision of quality medical care, and that if economic criteria are used, the criteria chosen truly reflect quality of care. PMID- 9398769 TI - Key issues in emergency medicine workforce planning. AB - The goal of workforce planning should be to match the supply of providers with the nation's demand. Whatever workforce planning process evolves, it will be necessary for the specialty to portray accurately our workforce needs. Because this portrait requires a clear understanding of the development of and funding mechanisms for graduate medical education and specific data describing both the supply and demand for emergency physicians, we address these issues. PMID- 9398770 TI - Emergency medicine and the academic health center. AB - The last 3 to 5 years have witnessed a tremendous change in direction for the academic health center. The major trends in this regard are increased importance of managed care, cost consciousness, development of a clinical system, pressure to downsize graduate medical education, and increased competition. PMID- 9398771 TI - The future of the certification system in emergency medicine. AB - The heart of the specialty of emergency medicine, like all specialties and subspecialties, is training. The excellence in medical care that has accrued to the American public has proceeded from the belief that a well-defined and accredited program of education will produce the highest probability that a physician providing care will be competent. There is now a joint opportunity in emergency medicine to build a certification and recertification system that meets the criteria to provide the highest quality care for the public and to offer an efficient and effective system for the members of the specialty. PMID- 9398772 TI - Access, quality, and cost control in emergency medicine: can we have all three? A resident's perspective on the future of emergency medicine. AB - The triad of access, quality, and cost provides a useful framework for the discussion of health care reform. A quote from a recent review of the Oregon Health Plan illustrates the conflict between these three factors very well. "The administrators of the plan are realistic people; they once placed a sign on the wall: 'Cost, access, quality--pick any two." PMID- 9398773 TI - Scientific publication in emergency medicine: imprint on the future. AB - Intensified research efforts in emergency medicine have resulted in a unique body of knowledge based on investigations involving emergency patients and conducted by emergency medicine researchers. The success and effectiveness of emergency medicine publications in the future rely on adherence to rigorous standards for peer review and editorial evaluation, introduction of value-added innovations, creative applications for electronic publication of scientific materials, and an unyielding commitment to quality and integrity. PMID- 9398774 TI - Quantifying the scanty science of prehospital emergency care. AB - Research can produce false-positive results just as can diagnostic tests. Uncontrolled studies have a specificity of only 11%, versus 88% for randomized controlled trials (RCTs), which have been designed to minimize the bias of investigators toward a positive outcome. A search of all the scientific studies in Medicine since 1985 revealed 5,842 publications on prehospital EMS, but only 54 were RCTs (and therefore unlikely to produce false-positive results). By way of comparison, during the same time hundreds of RCTs have been conducted on major medical emergency conditions, and RCTs on even minor topics such as urticaria and constipation exceed the scientific database on all of EMS. Of the 54 EMS RCTs, 4 (7%) reported harm from the new therapy, and 74% reported no effect of the new therapy at all. Only 7 (13%) RCTs showing a positive outcome of the intervention were uncontradicted; of these only 1 examined a major outcome such as survival, and only 1 compared the intervention with a placebo and could therefore evaluate the efficacy of EMS itself. Because there is such a paucity of scientific support for EMS interventions and because monitoring of outcomes and adverse effects is so poor, a serious reexamination of EMS practice is indicated. PMID- 9398775 TI - Developing a foundation for the evaluation of expanded-scope EMS: a window of opportunity that cannot be ignored. AB - EMS systems are about to undergo a major transformation. Not only will the scope of EMS change, but many experts believe that it will dramatically expand. Some see the "expanded scope" as entailing relatively limited changes, whereas others consider them to be more broad. Although no agreement is evident about the definition for expanded-scope EMS, it is hoped that all EMS professionals can agree that it must be implemented in a manner that can be carefully evaluated to determine its effects on patients and EMS systems. We present a framework for evaluating the effect of expanded-scope EMS in the various types of systems that currently exist. Special consideration must be given to the indirect effects that system changes may have on survival from out-of-hospital cardiac arrest. Numerous issues will affect our ability to properly assess expanded-scope EMS. The basic research models necessary to assess the impact of system change are lacking. Few EMS systems consistently produce significant volumes of good systems research ... that is, there are few "EMS laboratories." Cost-effectiveness and issues surrounding the "societal value" of EMS remain essentially unstudied. Reliable scoring methods, severity scales, and outcome measures are lacking: and, it is ethically and logistically difficult to justify withholding the "standard of care" in an effort to understand the impact of EMS interventions. Despite all of these barriers, it is time to pay the price of doing methodologically sound evaluations that ensure the most optimal societal impact by the EMS systems of the future. PMID- 9398776 TI - Multiple options and unique pathways: a new direction for EMS? AB - The same forces transforming the health care delivery system also are reshaping EMS. The changing economic and organizational structures of the health services delivery system may predict how EMS systems will redesign themselves. We discuss one template for future EMS systems. PMID- 9398777 TI - Emergency medicine in population-based systems of care. AB - EDs and emergency physicians play a critical role in health care delivery- providing care to those with life-threatening conditions, as well as serving as provider of last resort to those without options for primary care. This diversity of functions provides unique opportunities for identifying important unmet community needs and developing solutions to address these needs. Future challenges for emergency medicine include assessing and improving the quality of care provided within the ED and identifying the role of the ED in systems of care. Health services research can help define the optimal functions of emergency medicine in enhancing population health. PMID- 9398778 TI - The "new VA": delivering health care value through integrated service networks. AB - In an effort to meet powerful societal and industry-wide forces of change, the Veterans Health Administration has initiated a fundamental reengineering of itself, and is currently undergoing an innovative transformation that is among the most profound of any organization in American history. PMID- 9398779 TI - Fifty years of defibrillation. PMID- 9398780 TI - Guidelines for evaluation of international emergency medicine assistance and development projects. AB - Interest in the development of the specialty of emergency medicine and of emergency health care systems has greatly increased worldwide in the last few years. The guidelines in this article were developed in an effort to assist others in design and evaluation of all types of emergency medicine projects. PMID- 9398781 TI - Partners in progress: joining together against impaired driving. PMID- 9398782 TI - Current research in alcohol. PMID- 9398783 TI - Commentary: missing in action--emergency medicine and the problem drinker. PMID- 9398784 TI - Incarceration of a gravid fibroid uterus. AB - The case of an incarcerated gravid fibroid uterus in a 32-year-old woman is presented. This rare complication of pregnancy is readily identified by specific symptoms, physical examination, and ultrasound findings. If the condition is undiagnosed and untreated, spontaneous abortion and preterm labor often occur. PMID- 9398785 TI - Successful use of propofol in refractory delirium tremens. AB - Alcohol withdrawal is a common problem encountered by emergency physicians, with delirium tremens (DT) as the extreme manifestation. DT is a true medical emergency. Although benzodiazepines are the mainstay of therapy, some patients require massive amounts to control their symptoms. We report the successful use of propofol for DT refractory to benzodiazepines in a 42-year-old alcoholic man. We briefly discuss alcohol withdrawal, as well as the pharmacokinetics and adverse affects of propofol. The use of propofol in treating DT refractory to benzodiazepines has previously not been reported. PMID- 9398786 TI - Treatment of methanol poisoning with intravenous 4-methylpyrazole. AB - Treatment of human methanol poisoning with the alcohol dehydrogenase inhibitor, 4 methylpyrazole (fomepizole), has not been previously described. We report the clinical and toxicokinetic data of a patient with methanol poisoning who was treated with fomepizole. Formic acid levels remained undetectable during fomepizole treatment, the toxic effects of methanol were prevented, and the patient made a full recovery. PMID- 9398787 TI - "I'm sorry doctor! I'll just take the pain!". PMID- 9398788 TI - Frontline footage. PMID- 9398789 TI - Alarming defibrillator headlines. PMID- 9398790 TI - Treatment of panic disorder in the ED. PMID- 9398791 TI - Panic disorders, hyperventilation, and the dreaded brown paper bag. PMID- 9398792 TI - Acute toluene ingestion toxicity. PMID- 9398793 TI - Evaluation of gastric emptying function in clinical practice. AB - In this retrospective analysis, we compared different methods to evaluate gastric emptying function, aiming to improve the sensitivity and the clinical availability of our diagnostic testing. In the first study, we compared, in 72 patients clinically suspected of gastroparesis, the emptying of a meal containing two solid nutrients with different disintegration rates: 111In-labeled scrambled eggs and 99Tc-labeled liver cubes. Gastric emptying of 111In-labeled egg was delayed in 12 of our patients and the evacuation of the 99Tc-labeled liver was prolonged in 19 patients. The choice of the nutrient was not important for the identification of diabetic gastroparesis (43% vs 57%; NS), but it was determinant in the case of patients suspected of idiopathic gastroparesis (12% were positive with the egg and 25% with the liver; P < 0.05). In the second study, we compared two different diagnostic methods in 46 patients: a simple radiological detection of the gastric emptying of radiopaque pellets, and the scintigraphic emptying of a solid meal containing 99Tc-labeled liver cubes. Both tests correlated perfectly in 78% of our patients. In 15% of the population (six of these seven patients were diabetics suspected of gastroparesis) the scintigraphic method was normal, while the evacuation of radiopaque pellets was delayed. For clinical purposes, we therefore propose: (1) the scintigraphic method should use liver rather than egg as a radiolabeled tracer in order to improve the sensitivity of the test for detection of gastroparesis; and (2) the radiological detection of radiopaque markers is a reliable and convenient method for the detection of gastroparesis in clinical practice. It is possibly more sensitive than scintigraphy. PMID- 9398795 TI - Effects of biofeedback therapy on anorectal function in obstructive defecation. AB - Biofeedback therapy improves symptoms in patients with constipation and obstructive defecation. Whether it also improves anorectal function is unclear. Our purpose was to investigate prospectively the effects of biofeedback therapy on subjective and objective parameters of anorectal function in 25 consecutive patients with obstructive defecation. Biofeedback therapy consisted of pelvic floor relaxation exercises (phase I) and neuromuscular conditioning of rectal sensation and rectoanal coordination, with a solid state manometry system and simulated defecation maneuvers (phase II). The number of sessions was customized for each patient. Clinical improvement was assessed from the changes in anorectal manometry, balloon (50 cc) expulsion test, and the symptom and stool diaries. The number of therapy sessions varied [mean (range) = 6 (2-10)]. After therapy, when straining as if to defecate, the percentage anal relaxation, intrarectal pressure, and defecation index increased (P < 0.001). The balloon expulsion time, laxative consumption, and straining effort decreased (P < 0.001). Before therapy, 16/25 (64%) patients had impaired rectal sensation, and after therapy this improved (P < 0.001). After therapy, 15/25 (60%) patients reported > or = 75% satisfaction with bowel habit and 8/25 (32%) reported > or = 50% satisfaction (P < 0.001); 15/16 (94%) patients discontinued digital disimpaction. Biofeedback therapy not only improves subjective but also objective parameters of anorectal function in at least 76% of patients by rectifying the underlying pathophysiologic disturbance(s). Sensory conditioning and customizing the number of sessions may offer additional benefits. PMID- 9398794 TI - Differential responsiveness to contractile agents of isolated smooth muscle cells from human colons as a function of age and inflammation. AB - To study the involvement of age and inflammation in motor colonic activity in man, contractile responses to CCK, carbachol, and KCl of isolated colonic smooth muscle cells (SMC) from normal and inflamed human colons were evaluated; the incidence of sex and smoking on contraction was also analyzed. Contractile responses to the three agonists were significantly lower in tissues with a low degree of inflammation than in tissues with high level of inflammation or normal tissues. This reduction in cell responsiveness appears to be nonspecific and nonreceptor mediated. A positive correlation of the contractile responses to the three stimulants with the age of patients was observed. In contrast, no association was found between sex, smoking, and cell contraction. In conclusion, contractions of SMC due to CCK, carbachol, and KCl were significantly modified during life; inflammation of the colon led to a loss of SMC responsiveness. PMID- 9398796 TI - Hyperglycemia-induced attenuation of rectal perception depends upon pattern of rectal balloon inflation. AB - This study investigated the effects of acute hyperglycemia on conscious rectal perception in response to two different rectal distension paradigms. Eleven healthy males were studied in random order on two separate days during euglycemia and hyperglycemia with blood glucose concentrations clamped to 3.8 +/- 0.6 and 14.8 +/- 0.86 mmol/liter, respectively. In order to evoke sensory responses, rapid phasic and ramplike distensions were applied to an intrarectal balloon. Rectal sensation thresholds for initial sensation, sensation of stool and discomfort, and sensory intensities were recorded. Additionally, anorectal motor responses were investigated during phasic distension. Acute hyperglycemia did not modify rectal sensory pressure thresholds and perception scores in response to phasic distension. Neither did hyperglycemia alter the resting anal sphincter pressure, the pressure threshold for eliciting the rectoanal inhibitory reflex, or the maximal anal squeeze pressure. In contrast, hyperglycemia attenuated rectal perception in response to ramplike distension. The pressure thresholds, 10.0 +/- 1.8 and 17.0 +/- 3.6 mm Hg for initial sensation and discomfort, respectively, during hyperglycemia were significantly higher than the corresponding thresholds of 4.4 +/- 1.4 and 11.4 +/- 1.9 mm Hg observed during euglycemia (P < 0.01). Higher rectal pressures were observed at all intensities of sensation of stool and discomfort during hyperglycemia than those obtained during euglycemia (P < 0.01). Hyperglycemia did not alter the compliance of the rectum. The results of this study demonstrate that acute hyperglycemia attenuates rectal perception, and this attenuation depends upon the type of distension employed. Our findings also demonstrate that anal sphincter motor function is not appreciably modified by hyperglycemia. PMID- 9398797 TI - Adrenergic denervation hypersensitivity in ileal circular smooth muscle after small bowel transplantation in rats. AB - Effects of small bowel transplantation (SBT) on ileal motility are unknown. The aim of the present study was to investigate changes in spontaneous contractile activity and sensitivity to cholinergic and adrenergic agents in the ileal circular muscle after SBT in rats. Orthotopic SBT was performed in syngeneic rats to avoid immune phenomena. Distal ileal circular muscle strips from rats one week (N = 10) and eight weeks (N = 10) after SBT were stretched to optimal length (Lo), and basal spontaneous activity at Lo was measured. Dose-response experiments to the cholinergic agonist bethanechol (Be, 10(-8)-10(-4) M) were performed in the presence of tetrodotoxin (TTX, 10(-6) M) and to the adrenergic agonist norepinephrine (NE, 10(-8)-10(-4) M) with or without TTX. ED50 (negative log of drug-concentration that induced 50% effect) was calculated. We also studied rats with selective jejunoileal ischemia/ reperfusion, intestinal transection/reanastomosis, naive controls, and sham operated controls (N > or = 8/group). Spontaneous basal activity did not differ among groups. Sensitivity to Be was not different in rats after SBT or in other groups compared to control tissue. After SBT, hypersensitivity to NE was shown by a significant increase of ED50 at one and eight weeks after SBT (5.1 +/- 0.3 vs 6.2 +/- 0.4 and 6.2 +/- 0.2, respectively; P < 0.05) regardless of the presence of TTX. No hypersensitivity was observed after ischemia-reperfusion intestinal transection reanastomosis, or sham operation. It is concluded that ileal hypersensitivity to NE was related to the extrinsic denervation obligated by the transplantation procedure, possibly mediated through an increase in number of receptors on smooth muscle, not on the enteric nerves. PMID- 9398798 TI - Dual-channel ambulatory esophageal pH monitoring. A useful diagnostic tool? AB - Ambulatory pH monitoring of the distal esophagus is the most accurate diagnostic study for patients with suspected gastroesophageal reflux disease (GERD). The measurement of proximal esophageal acid exposure time may be useful in patients with atypical reflux symptoms. The aim of this study is to evaluate if proximal esophageal pH monitoring provides useful information beyond that learned with distal esophageal pH monitoring. We routinely performed dual-channel pH monitoring with pH electrodes positioned at 20 and 5 cm above the manometric lower esophageal sphincter from January 1992 to August 1995. All patients scored their esophageal symptoms from zero (none) to four (severe). We compared proximal esophageal reflux (PR) in patients with typical symptoms (i.e., heartburn, regurgitation) and in patients with atypical symptoms (i.e., chest pain, cough, hoarseness, and asthma). We compared symptom profiles between patients with and without PR. We reviewed our experience in patients with abnormal PR, but with a normal amount of distal esophageal reflux (DR). We studied 441 consecutive patients. There were no significant differences in PR between patients with typical and atypical symptoms. There were no differences in symptom profiles between patients with normal and abnormal PR. There were no differences of PR between the different atypical symptoms. PR did not correlate with the severity of the patient's symptoms. PR correlated well only with DR. Twenty-four patients had isolated abnormal PR, but only six patients improved with antireflux therapy. We conclude that routine ambulatory esophageal pH monitoring of the proximal esophagus appears to be of little value. The decision to offer patients an empiric trial of antireflux therapy for suspected GERD should not be based on the presence or absence of PR. PMID- 9398799 TI - Receiver operator characteristic analysis of endoscopy as a test for gastritis. AB - Endoscopic evaluation of the presence or absence of gastritis is often performed in lieu of biopsy and histologic diagnosis. The purpose of our study was to assess the value of endoscopic examination as a diagnostic test for gastritis. Two endoscopists prospectively assessed the antrum of 73 patients undergoing upper gastrointestinal endoscopy and graded, on a scale of 0-4 (0 = completely absent, 4 = definitely present), the likelihood of gastritis. The following features were also assessed at the time of endoscopy: erythema, nodularity, erosion, edema, and friability. Two concomitant antral biopsies (3 cm from the pylorus on the greater curvature of the stomach) were performed regardless of the endoscopic impression. The histologic findings were graded independently on a scale of 0-3 by two pathologists who were not aware of the endoscopic findings. The following histologic features were graded: acute inflammation, chronic inflammation, lymphoid aggregates, intestinal metaplasia, and quantity of Helicobacter pylori organisms. Receiver operator characteristic analysis, a method derived from signal detection theory, assesses the trade-off of sensitivity and specificity over all cutoff points of a test and is considered the best method by which to compare tests and determine the diagnostic utility of a given test. Receiver operator characteristic analysis gave an area of 0.65 +/- 0.01 SE for endoscopy as a test for gastritis (0.5 = chance, 1 = perfect) as defined by the histologic presence of inflammation. Additionally, endoscopy as a test for the presence of histologically proven Helicobacter pylori gave an area of 0.55 +/- 0.01 SE. All endoscopically graded features treated as separate tests for gastritis and/or H. pylori gave areas of approximately 0.44-0.61, indicative of a poor test. While H. pylori was always associated with at least some degree of inflammation, linear regression analysis revealed no correlation among any of the histologic features or of any histologic feature with any endoscopic feature. We conclude that a tissue diagnosis is essential for the proper diagnosis of gastritis. PMID- 9398800 TI - Differential effects of deoxycholic acid on proliferation of neoplastic and differentiated colonocytes in vitro. AB - The secondary bile acid deoxycholic acid is believed to be a promoter of large bowel cancer, in part by inducing colonic epithelial proliferation. The effects of deoxycholic acid on [3H]thymidine incorporation by the human colon cancer cell line HT29 and two differentiated subclones were measured and compared. The subclone HT29-C1 has features of mature absorptive cells and HT29-N2 cells secrete mucus under cholinergic control. The three cell lines were treated with deoxycholic acid (DCA) at concentrations of 0, 5, 10, 50, 100, 150, and 300 microM for 3, 6, 9, 15, 24, and 48 hr. A significant increase in proliferation was noted in HT29 cells only at 6 hr with 5 and 10 microM deoxycholic acid. Neither the subclone HT29-C1, nor HT29-N2 cells exhibited significant change in [3H]thymidine incorporation with DCA at these concentrations or time points. Higher doses of deoxycholic acid above 50 microM and duration of exposure greater than 24 hr were cytotoxic to all three cell lines. The proliferative effects of DCA in HT29 cells were not paralleled by changes in protein kinase C activity or protein kinase C isoform expression. Quantitative and qualitative differences in PKC isoform expression were not noted in the three cell lines used in this study. The proliferative effects of DCA on HT29 cells appear to be independent of the PKC signal transduction pathway. PMID- 9398801 TI - Primary hepatic high-grade non-Hodgkin's lymphoma and chronic hepatitis C infection. AB - Little is known about the coincidence of hepatitis C virus infection (HCV) and non-Hodgkin's lymphoma, although there is an increased incidence of chronic HCV infection with cryoglobulinemia type II and, interestingly, low-grade non Hodgkin's lymphoma (NHL) in a few patients. We therefore report on a 74-year-old white male with known chronic hepatitis C virus infection who was admitted to the clinic due to weight loss and pain in the right upper quadrant. Ultrasound examination was performed for suspected hepatocellular carcinoma since a lesion in the left lobe of the liver was seen. X-ray of the lungs showed a few scattered lesions, suggestive of metastases. The ultrasound-guided fine-needle puncture revealed a high-grade malignant B-cell NHL While alpha-fetoprotein was normal, both cryoglobulin type II and the polymerase chain reaction (PCR) for HCV were positive. After six cycles of chemotherapy consisting of CHOP, the patient showed complete remission over three years. Ultimately, he died due to a sudden myeloic blast crisis. In summary, we discuss the possible etiopathologic role of the hepatitis viruses in the occurrence of non-Hodgkin's lymphoma. As we and others showed that HCV infects peripheral mononuclear blood cells (PBML), the infected PBML not only may be a source for reinfection after orthotopic liver transplantation, but also could be the cause for transformation and monoclonal propagation of lymphomatous tissue. PMID- 9398802 TI - Serologic assay for secretory component distinguishes mechanical from hepatocellular cholestasis in humans. AB - In rats, serum secretory component (SC) is elevated in mechanical but not hepatocellular cholestasis. To determine if serum SC might distinguish cholestatic syndromes in humans, serum samples were obtained from control subjects and patients with mechanical and hepatocellular cholestasis. Equal volumes of serum were assayed for SC by immunoblotting with an antibody specific for human SC. Quantitative densitometry of these immunoblots showed that in mechanically obstructed patients serum SC was reversibly elevated to a level approximately 10-fold higher than that of patients with hepatocellular cholestasis (P < 0.001). When comparing the two cholestatic groups, levels of serum alkaline phosphatase, but not bilirubin and alanine aminotransferase, were significantly higher in the group with mechanical cholestasis (P < 0.01). When comparing individual patients, serum SC was more reliable than alkaline phosphatase in distinguishing the two cholestatic syndromes (P < 0.05). Thus, serum SC may distinguish mechanical from hepatocellular cholestasis in humans. PMID- 9398803 TI - Effects of somatostatin (SMS) on pancreatic microcirculation. AB - The effect of bolus infusion of increasing somatostatin (SMS) concentrations (1, 10, 100, 200 micrograms/100 g body wt) on pancreatic microcirculation and pancreatic tissue PO2 were investigated by using in vivo epifluorescence microscopy and a polarographic PO2 measurement technique. Additionally, the microperfusion of the pancreas, liver, spleen, stomach, and duodenum was measured by a laser Doppler device. Bolus infusion of SMS caused a significant, transient, and dose-dependent decrease in pancreatic capillary RBC velocities (to 50% of baseline) and acinar capillary overall perfusion (to 20% of baseline), which was not caused by a macrocirculatory depression. This pronounced decrease in microperfusion was not paralleled by a decline in tissue PO2. Laser Doppler measurements revealed that pancreatic and gastric microperfusion were reduced only at maximal SMS concentrations, considering that microperfusion of the liver, spleen, and duodenum was not altered. Therefore, we found further evidence that circulatory adjustment might occur during SMS inhibited secretory activity of the exocrine pancreas. PMID- 9398804 TI - Frequent detection of hepatitis B virus X-gene DNA in hepatocellular carcinoma and adjacent liver tissue in hepatitis B surface antigen-negative patients. AB - Hepatitis B virus is associated with human hepatocellular carcinoma. We performed polymerase chain reaction for the X, C, S, and preS2/S regions of the viral genome in 23 hepatitis B surface antigen-negative hepatocellular carcinomas and adjacent liver. Hepatitis B viral genomes were detected in 17 of 23 tumors and adjacent tissues (73.9%). Among recognized transactivators, the X gene was present in 16 (69.6%) cases of hepatocellular carcinoma, but preS2/S was detected in only 7 (30.4%). Hepatitis B virus C and S regions were detected in 3 (13.0%) and 9 (39.1%) hepatocellular carcinomas, respectively. Serologic study revealed antibodies to hepatitis B surface antigen, hepatitis B core antigen, and hepatitis B e antigen in 14 patients; among these, X-gene DNA was detected in 12 of 14 tumors (85.7%). The X gene was also detected in 4 of 9 tumors of seronegative patients. The X gene, present in many hepatocellular carcinomas, may promote hepatocellular carcinoma in hepatitis B surface antigen-negative patients. PMID- 9398805 TI - Prediction of effect of interferon on chronic hepatitis C. AB - Clinical, pathological, and virological analysis including hypervariable region-1 of hepatitis C virus (HCV) was performed to predict the effect of interferon (IFN) on 41 patients with chronic hepatitis type C. The low virus load, low frequency of the mutation in the hypervariable region-1 as the change of amino acid and high level of serum aminotransferase make one estimate the good effect of IFN on patients with HCV. Mutation in the hypervariable region-1 of HCV measured by fast assay fluorescence single-stranded conformational polymorphism was more frequent in nonresponders to IFN than responders. The most frequently mutated position was amino acid number 406. This indicates that the specific mutation site might affect the response of IFN. PMID- 9398806 TI - Increased levels of soluble adhesion molecules in the serum of patients with hepatitis C. Correlation with cytokine concentrations and liver inflammation and fibrosis. AB - Lymphocyte adhesion to endothelium, extravasation, and adhesion to hepatocytes are mediated by adhesion molecules and constitute important steps in the liver inflammation due to chronic hepatitis C (HCV-CH). We measured soluble intercellular adhesion molecule (sICAM-1, sCD54), vascular cell adhesion molecule (sVCAM-1, sCD106), E-selectin (sCD62E), as well as interleukin (IL)-1 beta, IL-8, and tumor necrosis factor-alpha (TNF-alpha) concentrations in the serum of 22 patients with HCV-CH in comparison to 20 seronegative healthy volunteers. sICAM 1, sVCAM-1, sCD62E, TNF-alpha, and IL-8 but not IL-1 beta concentrations were significantly elevated in patients. sICAM-1 and sCD62E correlated with TNF-alpha and aspartate amino transferases levels. sICAM-1 correlated with liver lobular inflammation whereas sVCAM-1, sCD62E, and IL-8 correlated with liver fibrosis. Measurement of soluble adhesion molecules may be an easy way to follow liver inflammation and fibrosis during HCV-CH. PMID- 9398807 TI - Hepatitis C serotypes in nonalcoholic and alcoholic patients. AB - Genotyping of the hepatitis C virus (HCV) RNA can be performed by a variety of methods following polymerase chain reaction amplification of a stable RNA portion of the genome. The gold standard is amplification of the RNA from the NS5 region, followed by direct sequencing and homology comparison. This method is extremely labor intensive. In this study, we compared an immunoblot serotyping technique (HCV SIA) to a reverse-hybridization line-probe assay (LiPA) for genotype classification among non-alcoholic HCV infected patients. We then compared and contrasted the response in this cohort to a population of alcoholic patients with HCV infection. To validate the serotype assay, sera from 110 patients with chronic HCV infection was utilized. Serotyping (Chiron SIA) and genotyping by the LiPA (Line Probe Assay, Innogenetics) reverse-hybridization technique was performed. Additionally, both methods were compared to sequence-derived genotyping in 26 patients based on PCR amplification of the NS5 region. After the validation phase, sera from 105 alcoholic patients was genotypically classified by the serologic method. The nonalcoholic and alcoholic groups were then compared with regard to serotype, demographics, and frequency of untypable test results. Among typable pairs, the overall concordance rate between serotyping and LiPA based genotyping was 93.75%. Patients with genotype 1 by reverse hybridization demonstrated a 95.8% concordance with serotype. Untypable samples were present for both techniques, but since they occurred in different patients, the techniques were complementary. Alcoholic patients were significantly more likely to be infected with untypable serotypes than those without a pattern of alcohol abuse. These patients were also more likely to be HCV RNA negative than sera from typable patients. Serotype 1 was associated with high HCV RNA titer and poor interferon treatment response among both nonalcoholic and alcoholic patients. An immunoblot method for the evaluation of genotype classification was rapid and easily performed compared to sequence-based genotyping. There was a high degree of concordance compared to reverse-hybridization and sequence-based genotype characterization methods. Failure to detect HCV RNA in the serum is associated with a higher likelihood of classification failure. This problem was particularly prevalent in the alcoholic population. HCV RNA titers and treatment outcomes were strongly associated with serotype classification results, demonstrating clinical utility of this assay technique. PMID- 9398808 TI - Hepatic denervation ameliorates sodium and water retention in experimental cirrhosis in rats. AB - Increased activity in the hepatic sympathetic nervous system may exacerbate salt and water retention in patients with liver cirrhosis. The aim of this study was to evaluate sodium and water homeostasis in rats with cirrhosis induced by diethylnitrosamine and to investigate the influence of hepatic denervation in this model. Animals were randomized into three groups: diethylnitrosamine-treated rats with (N = 13) and without (N = 8) hepatic denervation and control rats (N = 8). Rats were fed a normal salt diet (0.23% sodium ad libitum). The 24-hr measurements for sodium balance, water balance, and creatinine clearance were performed every two weeks for 12 weeks after surgery. Diethylnitrosamine-induced cirrhosis was confirmed histologically. The cumulative change in sodium balance in the innervated diethylnitrosamine-treated rat increased progressively and was significantly higher than the control during the last four weeks of the study. Meanwhile, rats with hepatic denervation showed significantly smaller changes in cumulative sodium balance at week 12 than those in the innervated group. The cumulative changes in water balance in the innervated group were significantly greater at weeks 10 and 12 than those of the denervated and control group, which remained unchanged throughout the study. Creatinine clearance in the innervated group decreased at weeks 10 and 12 by approximately 70% from baseline (P < 0.05); in contrast, it did not change significantly in the denervated group and control group throughout the study. These results demonstrated that hepatic denervation ameliorates sodium and water retention as well as glomerular function in cirrhosis model in rats. PMID- 9398809 TI - Touch cytology. A reliable and cost-effective method for diagnosis of Helicobacter pylori infection. AB - A variety of reliable methods are available for detecting Helicobacter pylori (Hp) during upper gastrointestinal endoscopy. We evaluated the clinical utility and cost-effectiveness of rapid urease test (RUT), touch cytology (TC), and histology (H). Two hundred thirty-eight consecutive patients (178 without previous medical treatment and 60 formerly treated with anti-Hp therapy) were tested for Hp infection by RUT, TC, and H (H&E stain). The infection status for each patient was established by a concordance of two test results. The time to carry out the three tests and their cost were also calculated. Sensitivity of TC (100%) was significantly higher than that of RUT (86.8%; P < 0.001), but not than that of H (94.9%). RUT was significantly more specific than H (100% vs 95.6%; P < 0.05), but not than TC (96.4%). Hp infection was more frequent in the patients with chronic active gastritis than in those with chronic nonactive gastritis (P < 0.001). No Hp infection was detected in absence of chronic antral inflammation. RUT resulted the cheapest method and H the most expensive; TC is faster and cheaper than H. When additional information about the severity of mucosal damage or the presence of cell atypias is not necessary, histologic examination can be omitted, and a cost-effective strategy for assessing Hp status might consist in taking two antral biopsies, the former for performing RUT and the latter for preparing a slide by TC, which should be stained and examined only when the RUT result is negative. PMID- 9398810 TI - Effect of omeprazole 40 mg once daily on intraduodenal and intragastric pH in H. pylori-negative healthy subjects. AB - There is a lack of information about the effect of omeprazole or other antisecretory drugs on intraduodenal pH. Aim of the study was to document the variation over time of intraduodenal pH during a 24-hr period and to simultaneously study the effect of omeprazole 40 mg once daily on intragastric and intraduodenal pH in healthy H. pylori-negative subjects. In a randomized, placebo-controlled study, eight subjects (five women, three men, mean age 22.7 years) received oral 40 mg omeprazole or placebo once daily for eight days. On day 7, intragastric and intraduodenal pH was measured continuously for 24 hr, using two miniature glass-membrane electrodes placed in the stomach (fundus) and in the distal third of the duodenum. The 24-hr median intraduodenal pH was 5.95 with placebo and 5.85 with omeprazole. Median intragastric pH was 1.68 without and 4.93 with omeprazole. During omeprazole therapy, intragastric pH fell below 4.0 in five of eight subjects. In the 2- and 3-hr postprandial periods, the percentage of time with pH < 5 was significantly reduced with omeprazole. In healthy subjects, 24-hr median and postprandial pH in the distal part of the duodenum was lower than previously thought. Omeprazole significantly reduced the percentage of time with pH < 5 postprandially. At night, intragastric pH fell below 4.0 with omeprazole 40 mg once daily. Omeprazole does not change 24-hr median intraduodenal pH significantly. PMID- 9398811 TI - Is electrogastrography a substitute for manometric studies in children with functional gastrointestinal disorders? AB - We performed simultaneous fasting and fed antroduodenal manometry and EGG in 25 children with functional bowel disorders. Three patients (12%) had an uninterpretable EGG. The manometric studies showed severe neuropathy in six patients; milder neuropathic changes in five patients; postprandial hypomotility in one patient; myopathy in four patients, and normal motility in the remaining six patients. The percentage of tachygastria time (frequency > 3.5 cycles/min) was higher in the patiens with mild (44.1 +/- 15.8%) and severe (48 +/- 19.1%) neuropathy than in the patients with myopathy (20 +/- 16.2%, P < 0.05) or with normal motility (23 +/- 13.3%, P < 0.05). There was a considerable overlap in the percentage of tachygastria and total arrhythmia time among the different study groups. The ratio of post- to preprandial power was significantly higher (2.5 +/- 0.07) in children with normal motility than in the other patients groups. Every child with total arrhythmia time < 35% and a ratio of post- to preprandial power > 2.4 had normal manometry. In summary, EGG differentiated groups of children with normal manometry from others with neuropathic or myopathic changes, but in a minority of patients the study was not interpretable and there was overlap in EGG results between children with normal and abnormal manometry. PMID- 9398812 TI - Impedance planimetric characterization of esophagus in systemic sclerosis patients with severe involvement of esophagus. AB - This study was designed to evaluate the distensibility and secondary peristalsis of the esophagus in patients suffering from systemic sclerosis with severe esophageal involvement. Balloon distension with impedance planimetric measurement of luminal cross-sectional area was done 7 and 15 cm above the lower esophageal sphincter in 13 patients and nine healthy controls. The controls were studied both with and without receiving the anticholinergic drug butylscopolamine. The cross-sectional area--pressure relations were nonlinear with the largest cross sectional area in patients at both measuring sites when compared to controls (P < 0.001). The anticholinergic drug butylscopolamine increased the cross-sectional area in controls (P < 0.001). The cross-sectional area distensibility, defined as CSA0(-1) delta CSA delta P-1 did not differ between patients and controls. Balloon distensions elicited contractions proximal to the distension site. The amplitude and frequency of contractions at the distal distension site were significantly reduced in the patients when compared to the controls (P < 0.05). In conclusion, the distal esophagus is most severely affected in patients with systemic sclerosis with increased cross-sectional area and impaired peristalsis. PMID- 9398813 TI - Dyspepsia due to eosinophilic gastroenteritis. AB - Classical eosinophilic gastroenteritis is a rare disease but may be misdiagnosed in clinical practice. We report eosinophilic gastroenteritis that was diagnosed in six patients (four males and two females; mean age 31.5 years) using standard criteria (presence of gastrointestinal symptoms, a predominant eosinophilic infiltrate on biopsy, and exclusion of other causes of eosinophilia). All had gastric mucosal disease and presented with dyspepsia. The median duration of symptoms prior to diagnosis was three months (range five weeks to 13 years). Epigastric pain or discomfort was the most common symptom (100%) followed by anorexia, nausea, and vomiting (67%, 67% and 33%, respectively). None had diarrhea. Half the patients had a history of allergy, while 67% had peripheral eosinophilia. All responded to oral steroids within two months; one third needed to continue on a small dose of maintenance steroids to remain in remission. A high degree of suspicion and biopsy at upper endoscopy is necessary for diagnosis of this rare disease. PMID- 9398814 TI - The place of upper gastrointestinal tract endoscopy before and after vertical banded gastroplasty for morbid obesity. AB - In industrialized countries, surgical gastroplasty is performed more and more frequently in patients with morbid obesity. The aims of this prospective study were to determine the incidence of upper gastrointestinal lesions in obese patients and to assess the place of digestive endoscopy in symptomatic patients after gastroplasty. A consecutive group of 159 obese patients were studied before and after vertical banded gastroplasty. In the preoperative evaluation, reflux esophagitis and gastroduodenal lesions were endoscopically observed in 31% and 37% of the patients, respectively. Interestingly, the majority of the obese patients with upper gastrointestinal lesions were asymptomatic. In the postoperative follow-up period, 55 of the 159 patients complained of upper gastrointestinal symptoms such as vomiting (72%), esophageal reflux (17%), and epigastric pain (3%). Stenosis of the outlet of the gastric pouch was described in 40 of the 55 symptomatic patients. Esophagitis was observed in 60% of these patients. Endoscopic dilation using Savary bougies or TTS balloon was successfully performed in all the patients with symptomatic stenosis of the gastric outlet. Food impaction was endoscopically removed in four patients. Thus, we recommend performing an upper gastrointestinal endoscopy in obese patients who are candidates for surgical gastroplasty because of the high incidence of upper gastrointestinal peptic lesions. Endoscopy is also helpful in patients with digestive disorders occurring after gastroplasty in order to define and to treat the lesions. PMID- 9398815 TI - Peripheral blood lymphocyte beta 2 integrin and ICAM expression in inflammatory bowel disease. AB - The beta 2 integrin intercellular adhesion molecule (ICAM) adhesion pathway is likely pivotal in the immunopathogenesis of inflammatory bowel disease (IBD). We have undertaken a comprehensive study of peripheral blood lymphocyte (PBL) expression of all beta 2 integrins and ICAMs in patients with IBD using flow cytometry and assessed our data on the basis of IBD diagnosis, disease state of activity, and use of corticosteroids. Blood was collected from patients with Crohn's disease (N = 49), ulcerative colitis (N = 43), and normal control volunteers (N = 15). Mononuclear cells were separated using a Ficoll-Hypaque gradient and prepared for flow cytometry. The data were analyzed for percentage expression, mean fluorescent intensity (MFI) as well as for histogram patterns. The analysis was stratified for disease diagnosis, disease activity level, and for use of prednisone among patients with active disease. There was decreased percentage expression of CD11a, CD18, and ICAM-3 in Crohn's disease and ulcerative colitis compared with normal, but an increased MFI for these molecules among patients with Crohn's disease. Active Crohn's disease showed a greater change in this pattern compared with both inactive disease and active ulcerative colitis. CD11a and CD18 histograms typically had two peaks of expression. The predominance of one peak over the other varied with disease diagnosis and activity. CD11b and alpha d expression patterns were not different in IBD compared with normal. CD11c was not expressed by PBLs and, ICAM-2, typically an endothelial ligand, was expressed on PBLs. There were changes in the expression of beta 2 integrins in IBD, which were more evident in Crohn's disease than ulcerative colitis. We hypothesize that the decreased percentage expression and increased MFI of CD11a, CD18, and ICAM-3 may suggest that cells up-regulate these ligands following activation and are egressing into tissue. PMID- 9398816 TI - Immunoglobulin G subclass distribution of anti-endothelial cell antibodies (AECA) in patients with ulcerative colitis or Crohn's disease. AB - Anti-endothelial cell antibodies have been described in sera from patients with inflammatory bowel disease. The aim of this study was to determine, by ELISA, the IgG subclass distribution of anti-endothelial cell antibodies, in patients with ulcerative colitis (N = 28) or Crohn's disease (N = 82) as compared with blood donors (N = 95). Thirty-six percent of ulcerative colitis and 23% of Crohn's disease patients were positive for at least one of the IgG anti-endothelial cell subclasses. Interestingly, the pattern of IgG anti-endothelial cell subclass observed in the two inflammatory bowel diseases differs. In Crohn's disease, the IgG1 anti-endothelial cell antibody level was significantly increased (P < 0.05) while IgG2 and IgG4 anti-endothelial cell antibody levels were decreased (P < 0.0001 and P < 0.01, respectively) as compared to ulcerative colitis patients. The immunoglobulin G3 anti-endothelial cell antibody level was decreased in both ulcerative colitis and Crohn's disease patients as compared to healthy blood donors. No relationship was detected between disease activity of ulcerative colitis or Crohn's disease patients and anti-endothelial cell IgG subclasses. Finally, the disparity of IgG anti-endothelial cell subclass distribution in these two inflammatory bowel diseases suggests that the ability to activate effector mechanisms is not identical, and hence, deals with the concept of distinctive pathogenetic mechanisms in these two diseases. PMID- 9398818 TI - Intractable Crohn's colitis and perianal disease responding to cyclophosphamide and epirubicin. AB - A case is reported where intractable Crohn's disease responded to chemotherapy for breast carcinoma. The drug most likely responsible was cyclophosphamide. Cyclophosphamide may have a role in the management of inflammatory bowel disease, but this needs to be confirmed before it can be recommended as a treatment option. PMID- 9398817 TI - Activation of plasma contact and coagulation systems and neutrophils in the active phase of ulcerative colitis. AB - We have shown that the contact (kallikrein-kinin) system is involved in the pathogenesis of experimental enterocolitis. We now investigate activation of the contact and coagulation pathways, platelets, and neutrophils in active and inactive ulcerative colitis patients as compared to normal controls. In active ulcerative colitis patients, a significant decrease of plasma prekallikrein, high molecular weight kininogen, and C1 inhibitor levels was observed as compared with controls, as well as prekallikrein activation on western blots. Significant elevation of prothrombin fragment (F1 + 2), which indicates thrombin generation, and elastase-alpha 1-antitrypsin complexes, reflecting neutrophil activation, were found in patients with active disease. Plasma beta-thromboglobulin, a marker of platelet activation, was elevated in both active and inactive disease and appears to be a feature of ulcerative colitis. Activation of contact and coagulation pathways, as well as neutrophils, may mediate inflammation in the active phase of ulcerative colitis. PMID- 9398819 TI - Modification of colonic fermentation by bifidobacteria and pH in vitro. Impact on lactose metabolism, short-chain fatty acid, and lactate production. AB - Colonic fermentation plays an important role in the prevention of lactose intolerance and intestinal disorders. The objectives of this study were to evaluate whether supplementation with bifidobacteria modify colonic fermentation of lactose and short-chain fatty acid production and to assess influence of the pH in an in vitro continuous culture system. There was a significantly greater reduction in lactose concentrations at pH 6.7 than that at either pH 6.2 or pH 5.7, accompanied by the highest beta-galactosidase activity and D-lactate production. Bifidus supplementation reduced lactose and D-lactate concentrations and increased acetate production at pH 6.7. The study demonstrates that lactose is rapidly metabolized by colonic bacteria and lactose fermentation in vitro is pH dependent with a maximum rate at pH 6.7. Bifidobacteria supplementation may have the potential to improve lactose fermentation and to manipulate SCFA and lactate production. PMID- 9398821 TI - Morphological characteristics, epithelial cell proliferation, and crypt fission in cecum and colon of growing pigs. AB - Morphological characteristics and cellular proliferation were investigated in the hindgut tissue of 25 pigs ranging from 5 to 261 days of age; the three youngest pigs were unweaned. Tissue samples were taken from the cecum and from the proximal, middle, and distal part of the colon. In the young pigs a high incidence of branched crypts was observed. During the first three to four months there was an increase in crypt height and proliferative activity, as determined by the mitotic count, as well as an increase in the mucin secretion, especially the sulfomucins. Distinct regional differences were observed between the four intestinal sites. In general, the crypts were deeper and more closely spaced and the turnover time was higher in the distal part of the colon as compared to the cecum and the proximal colon. Furthermore, a greater proportion of the mucins in the middle and distal part of the colon are acidic or sulfated as compared to the cecum, where the mucins are more of the neutral type. These regional and age related differences in morphological characteristics of the hindgut in pigs may have significance for the etiology of intestinal infections. PMID- 9398820 TI - Submandibular gland peptide-T (SGP-T) inhibits intestinal anaphylaxis. AB - A novel peptide, submandibular gland peptide-T (SGP-T), which reduces allergen induced hypotension, was examined for effects on intestinal anaphylaxis. Hooded Lister rats were sensitized to egg albumin and prepared for the measurement of in vivo myoelectric activity of the jejunum. The disruption of migrating myoelectric complexes (MMCs) that occurs upon intraluminal, duodenal challenge with antigen of sensitized rats was inhibited by 75% upon intravenous treatment with 100 micrograms/kg of SGP-T. In addition, SGP-T reduced the number of rats experiencing anaphylactic diarrhea and disrupted MMCs, but the peptide did not alter antigen-provoked release of rat mast cell protease II. The mechanism of action of SGP-T remains to be determined, but it apparently does not act directly on mast cells to exert its antianaphylactic action. These results emphasize that modulation of immediate hypersensitivity reactions is only one of several gastrointestinal activities that are affected by growth factors and peptides released from salivary glands. PMID- 9398822 TI - St. John's wort. PMID- 9398823 TI - Olopatadine for allergic conjunctivitis. PMID- 9398824 TI - Pramipexole and ropinirole for Parkinson's disease. PMID- 9398825 TI - Genetic(al) correctness. PMID- 9398826 TI - The genetics of air pollution. PMID- 9398827 TI - Sugar and spice and all things splice? PMID- 9398828 TI - Parsing age, mutations and time. PMID- 9398829 TI - Sounding out a novel sulphate transporter. PMID- 9398830 TI - Expanding on population studies. PMID- 9398831 TI - Haemochromatosis, HFE and genetic complexity. PMID- 9398832 TI - Cytosine methylation targetted to pre-determined sequences. PMID- 9398833 TI - Normal meiotic recombination in p53-deficient mice. PMID- 9398834 TI - Mouse loci for malaria-induced mortality and the control of parasitaemia. PMID- 9398835 TI - Genetic control of blood parasitaemia in mouse malaria maps to chromosome 8. PMID- 9398836 TI - Mutation in hepatocyte nuclear factor-1 beta gene (TCF2) associated with MODY. PMID- 9398837 TI - A CAG/CTG expansion in the normal population. PMID- 9398838 TI - A twin-pronged attack on complex traits. AB - Before one starts the hunt for quantitative trait loci (QTLs) for a complex trait, it is necessary to show that the trait is genetically influenced. This evidence is most likely to come from the classical twin study--the demonstration that monozygotic twins are more similar for the trait than dizygotic twins. The strengths and weaknesses of twin studies are discussed, and it is suggested that, far from becoming irrelevant with advances in molecular biology, they can improve the efficiency of QTL detection and play an important role in unravelling developmental genetic mechanisms. PMID- 9398839 TI - Positional cloning of the APECED gene. AB - Autoimmune polyglandular syndrome type I (APS 1, also called APECED) is an autosomal-recessive disorder that maps to human chromosome 21q22.3 between markers D21S49 and D21S171 by linkage studies. We have isolated a novel gene from this region, AIRE (autoimmune regulator), which encodes a protein containing motifs suggestive of a transcription factor including two zinc-finger (PHD finger) motifs, a proline-rich region and three LXXLL motifs. Two mutations, a C- >T substitution that changes the Arg 257 (CGA) to a stop codon (TGA) and an A-->G substitution that changes the Lys 83 (AAG) to a Glu codon (GAG), were found in this novel gene in Swiss and Finnish APECED patients. The Arg257stop (R257X) is the predominant mutation in Finnish APECED patients, accounting for 10/12 alleles studied. These results indicate that this gene is responsible for the pathogenesis of APECED. The identification of the gene defective in APECED should facilitate the genetic diagnosis and potential treatment of the disease and further enhance our general understanding of the mechanisms underlying autoimmune diseases. PMID- 9398840 TI - An autoimmune disease, APECED, caused by mutations in a novel gene featuring two PHD-type zinc-finger domains. AB - Autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED) is the only described systemic autoimmune disease with established monogenic background, and the first autoimmune disorder localized outside the major histocompatibility complex (MHC) region. The primary biochemical defect in APECED is unknown. We have isolated a novel gene, AIRE, encoding for a putative nuclear protein featuring two PHD-type zinc-finger motifs, suggesting its involvement in transcriptional regulation. Five mutations in AIRE are reported in individuals with this disorder. This is the first report of a single-gene defect causing a systemic human autoimmune disease, providing a tool for exploring the molecular basis of autoimmunity. PMID- 9398841 TI - Huntingtin is required for neurogenesis and is not impaired by the Huntington's disease CAG expansion. AB - Huntington's disease (HD) is an autosomal-dominant neurodegenerative disorder caused by a CAG repeat expansion that lengthens a glutamine segment in the novel huntingtin protein. To elucidate the molecular basis of HD, we extended the polyglutamine tract of the mouse homologue, Hdh, by targetted introduction of an expanded human HD CAG repeat, creating mutant HdhneoQ50 and HdhQ50 alleles that express reduced and wild-type levels of altered huntingtin, respectively. Mice homozygous for reduced levels displayed characteristic aberrant brain development and perinatal lethality, indicating a critical function for Hdh in neurogenesis. However, mice with normal levels of mutant huntingtin did not display these abnormalities, indicating that the expanded CAG repeat does not eliminate or detectably impair huntingtin's neurogenic function. Thus, the HD defect in man does not mimic complete or partial Hdh inactivation and appears to cause neurodegenerative disease by a gain-of-function mechanism. PMID- 9398842 TI - Pendred syndrome is caused by mutations in a putative sulphate transporter gene (PDS). AB - Pendred syndrome is a recessively inherited disorder with the hallmark features of congenital deafness and thyroid goitre. By some estimates, the disorder may account for upwards of 10% of hereditary deafness. Previous genetic linkage studies localized the gene to a broad interval on human chromosome 7q22-31.1. Using a positional cloning strategy, we have identified the gene (PDS) mutated in Pendred syndrome and found three apparently deleterious mutations, each segregating with the disease in the respective families in which they occur. PDS produces a transcript of approximately 5 kb that was found to be expressed at significant levels only in the thyroid. The predicted protein, pendrin, is closely related to a number of known sulphate transporters. These studies provide compelling evidence that defects in pendrin cause Pendred syndrome thereby launching a new area of investigation into thyroid physiology, the pathogenesis of congenital deafness and the role of altered sulphate transport in human disease. PMID- 9398843 TI - Tumorigenesis and a DNA repair defect in mice with a truncating Brca2 mutation. AB - Germline mutation of the BRCA2 gene carries a high risk of developing breast cancer. To study the function of this gene, we generated a mutation in Brca2 in mice. Unlike other mutations in the Brca2 gene, which are lethal early in embryogenesis when homozygous, some of our homozygous mutant mice survive to adulthood. These animals have a wide range of defects, including small size, improper differentiation of tissues, absence of germ cells and the development of lethal thymic lymphomas. Fibroblasts cultured from BrcaZ-/-embryos have a defect in proliferation that may be mediated by over-expression of p53 and p21Waf1/CIP1. We show that Brca2 is required for efficient DNA repair, and our results suggest that loss of the p53 checkpoint may be essential for tumour progression triggered by mutations in BRCA2. PMID- 9398844 TI - Rapid accumulation of genome rearrangements in liver but not in brain of old mice. AB - Somatic mutations have long been considered a possible cause of ageing. To directly study mutational events in organs and tissues of ageing mammals, a transgenic mouse model has been generated that harbours lacZ reporter genes as part of chromosomally integrated plasmids. Using this model, we determined spontaneous mutant frequencies and spectra in mouse liver and brain as a function of age. In the liver, mutant frequencies increased with age from birth to 34 months; in the brain, an increase was observed only between birth and 4-6 months. Molecular characterization of the mutations showed that a substantial portion involved genome rearrangement events, with one breakpoint in a reporter gene and the other in the mouse flanking sequence. In the liver, these genome rearrangements did not increase with age until after 27 months, when they increased rapidly. In brain, the frequency of genome rearrangements was lower than in liver and did not increase with age. PMID- 9398845 TI - Demographic history and linkage disequilibrium in human populations. AB - In the human genome, linkage disequilibrium (LD)--the non-random association of alleles at chromosomal loci--has been studied mainly in regions surrounding disease genes on affected chromosomes. Consequently, little information is available on the distribution of LD across anonymous genomic regions in the general population. However, demographic history is expected to influence the extent of overall LD across the genome, so a population that has been of constant size will display higher levels of LD than a population that has expanded. In support of this, the extent of LD between anonymous loci on chromosome 4 in chimpanzees (as a model of a population of constant size) has been compared to that in Finns (as a model of an expanded population; refs 8,9) and found to exhibit more LD than in the latter population. In Europe, studies of mitochondrial (mt) DNA sequences have suggested that most populations have experienced expansion, whereas the Saami in northern Fenno-Scandinavia have been of constant size (Table 1). Thus, in northern Europe, populations with radically different demographic histories live in close geographic proximity to each other. We studied the allelic associations between anonymous microsatellite loci on the X chromosome in the Saami and neighbouring populations and found dramatically higher levels of LD in the Saami than in other populations in the region. This indicates that whereas recently expanded populations, such as the Finns, are well suited to map single disease genes affected by recent mutations, populations that have been of constant size, such as the Saami, may be much better suited to map genes for complex traits that are caused by older mutations. PMID- 9398846 TI - Urokinase-generated plasmin activates matrix metalloproteinases during aneurysm formation. AB - The molecular mechanisms predisposing to atherosclerotic aneurysm formation remain undefined. Nevertheless, rupture of aortic aneurysms is a major cause of death in Western societies, with few available treatments and poor long-term prognosis. Indirect evidence suggests that matrix metalloproteinases (MMPs) and plasminogen activators (PAs) are involved in its pathogenesis. MMPs are secreted as inactive zymogens (pro-MMPs), requiring activation in the extracellular compartment. Plasmin, generated from the zymogen plasminogen by tissue-type plasminogen activator (t-PA) or urokinase-type plasminogen activator (u-PA; refs 14,15), has been proposed as a possible activator in vitro, but evidence for such a role in vivo is lacking. Analysis of atherosclerotic aorta in mice with a deficiency of apoliprotein E (Apoe-/-; ref. 18), singly or combined with a deficiency of t-PA (Apoe-/-:Plat-/-) or of u-PA (Apoe-/-:Plau-/-; ref. 19), indicated that deficiency of u-PA protected against media destruction and aneurysm formation, probably by means of reduced plasmin-dependent activation of pro-MMPs. This genetic evidence suggests that plasmin is a pathophysiologically significant activator of pro-MMPs in vivo and may have implications for the design of therapeutic strategies to prevent aortic-wall destruction by controlling Plau gene function. PMID- 9398847 TI - Mutations in PEX1 are the most common cause of peroxisome biogenesis disorders. AB - The peroxisome biogenesis disorders (PBDs) are a group of lethal autosomal recessive diseases caused by defects in peroxisomal matrix protein import, with the concomitant loss of multiple peroxisomal enzyme activities. Ten complementation groups (CGs) have been identified for the PBDs, with CG1 accounting for 51% of all PBD patients. We identified the human orthologue of yeast PEX1, a gene required for peroxisomal matrix protein import. Expression of human PEX1 restored peroxisomal protein import in fibroblasts from 30 CG1 patients, and PEX1 mutations were detected in multiple CG1 probands. A common PEX1 allele, G843D, is present in approximately half of CG1 patients and has a deleterious effect on PEX1 activity. Phenotypic analysis of PEX1-deficient cells revealed severe defects in peroxisomal matrix protein import and destabilization of PEX5, the receptor for the type-1 peroxisomal targetting signal, even though peroxisomes were present in these cells and capable of importing peroxisomal membrane proteins. These data demonstrate an important role for PEX1 in peroxisome biogenesis and suggest that mutations in this gene are the most common cause of the PBDs. PMID- 9398848 TI - Human PEX1 is mutated in complementation group 1 of the peroxisome biogenesis disorders. AB - Human peroxisome biogenesis disorders (PBDs) are a group of genetically heterogeneous autosomal-recessive disease caused by mutations in PEX genes that encode peroxins, proteins required for peroxisome biogenesis. These lethal diseases include Zellweger syndrome (ZS), neonatal adrenoleukodystrophy (NALD) and infantile Refsum's disease (IRD), three phenotypes now thought to represent a continuum of clinical features that are most severe in ZS, milder in NALD and least severe in IRD2. At least eleven PBD complementation groups have been identified by somatic-cell hybridization analysis compared to the eighteen PEX complementation groups that have been found in yeast. We have cloned the human PEX1 gene encoding a 147-kD member of the AAA protein family (ATPases associated with diverse cellular activities), which is the putative orthologue of Saccharomyces cerevisiae Pex1p (ScPex1p). Human PEX1 has been identified by computer-based 'homology probing' using the ScPex1p sequence to screen databases of expressed sequence tags (dbEST) for human cDNA clones. Expression of PEX1 rescued the cells from the biogenesis defect in human fibroblasts of complementation group 1 (CG1), the largest PBD complementation group. We show that PEX1 is mutated in CG1 patients. PMID- 9398849 TI - Atm selectively regulates distinct p53-dependent cell-cycle checkpoint and apoptotic pathways. AB - Atm is part of a pathway that responds to DNA damage from ionizing radiation (IR). This pathway involves p53, as Atm-deficient cell lines and mice are defective in p53 induction after IR. p53 is a multi-functional protein that simultaneously regulates distinct downstream pathways controlling cell-cycle progression and apoptosis. However, the mechanisms by which p53 differentially activates downstream pathways are unknown. To determine the relationship between Atm and p53, we examined cell-cycle and apoptotic responses in Atm-, p53-(ref. 8) and p21-deficient mice after IR in the whole animal. As expected, p53 protein levels were not induced by IR in thymus of Atm-deficient mice. IR-induced cell cycle checkpoint function was also defective, and induction of p21 was attenuated in thymus from Atm-deficient mice. However, IR-induced apoptosis and Bax induction were completely normal; both of which are mediated by p53. IR-induced thymic apoptosis was suppressed in Atm/p53 double-mutant mice but not in Atm/p21 double mutants, demonstrating p53 dependence and Atm independence. Thus, Atm deficiency results in lack of p53 induction by IR, but only selective disruption of p53-dependent functions. Our results support a model in which upstream effectors such as Atm selectively activate p53 to regulate specific downstream pathways, providing a mechanism for controlling distinct cell-cycle and apoptotic responses. PMID- 9398850 TI - ATM and RPA in meiotic chromosome synapsis and recombination. AB - ATM is a member of the phosphatidylinositol 3-kinase (PIK)-like kinases, some of which are active in regulating DNA damage-induced mitotic cell-cycle checkpoints. ATM also plays a role in meiosis. Spermatogenesis in Atm-/- male mice is disrupted, with chromosome fragmentation leading to meiotic arrest; in human patients with ataxia-telangiectasia (A-T), gonadal atrophy is common. Immuno localization studies indicate that ATM is associated with sites along the synaptonemal complex (SC), the specialized structure along which meiotic recombination occurs. Recombination, preceded by pairing of homologous chromosomes, is thought to require heteroduplex formation between homologous DNA, followed by strand exchange. These early meiotic steps (entailing the formation and processing of meiotic recombination intermediates with DNA-strand interruptions) require ssDNA-binding proteins such as replication protein A (RPA; refs 5-7). In somatic cells, DNA damage induces ATM-dependent phosphorylation of RPA. We demonstrate here that ATM and RPA co-localize along synapsed meiotic chromosomes and at sites where interactions between ectopic homologous chromosome regions appear to initiate. In Atm-/- meiotic prophase spermatocytes, immuno localization shows that RPA is present along synapsing chromosomes and at sites of fragmentation of the SC. These results suggest that RPA and ATM co-localize at sites where interhomologous-DNA interactions occur during meiotic prophase and where breaks associated with meiotic recombination take place after synapsis, implying a possible functional interaction between these two proteins. PMID- 9398851 TI - Partial rescue of the prophase I defects of Atm-deficient mice by p53 and p21 null alleles. AB - Patients with the human disorder ataxia-telangiectasia (A-T; refs 1,2) and Atm deficient mice have a pleiotropic phenotype that includes infertility. Here we demonstrate that male gametogenesis is severely disrupted in Atm-deficient mice in the earliest stages of meiotic prophase I, resulting in apoptotic degeneration. Atm is required for proper assembly of Rad51 onto the chromosomal axial elements during meiosis. In addition, p53, p21 and Bax are elevated in testes from Atm-deficient mice. To determine whether these elevated protein levels are important factors in the meiotic disruption of Atm-deficient mice, we analysed the meiotic phenotype of Atm/p53 or Atm/p21 double mutants. In these double mutants, meiosis progressed to later stages but was only partly rescued. Assembly of Rad51 foci on axial elements remained defective, and gametogenesis proceeded only to pachytene of prophase I. Previous results demonstrated that mice homozygous for a null mutation in Rad51 (ref. 6) display an early embryonic lethal phenotype that can be partly rescued by removing p53 and/or p21. Because Atm-deficient mice are viable but completely infertile, our studies suggest that the Rad51 assembly defects and elevated levels of p53, p21 and Bax represent tissue-specific responses to the absence of Atm. PMID- 9398852 TI - Donor splice-site mutations in WT1 are responsible for Frasier syndrome. AB - Frasier syndrome (FS) is a rare disease defined by male pseudo-hermaphroditism and progressive glomerulopathy. Patients present with normal female external genitalia, streak gonads and XY karyotype and frequently develop gonadoblastoma. Glomerular symptoms consist of childhood proteinuria and nephrotic syndrome, characterized by unspecific focal and segmental glomerular sclerosis, progressing to end-stage renal failure in adolescence or early adulthood. No case of Wilms' tumour has been reported, even in patients with extended follow-up. In contrast with FS patients, most individuals with Denys-Drash syndrome (DDS; refs 6,7) have ambiguous genitalia or a female phenotype, an XY karyotype and dysgenetic gonads. Renal symptoms are characterized by diffuse mesangial sclerosis, usually before the age of one year, and patients frequently develop Wilms' tumour. Mutations of the Wilms'-tumour gene, WT1, cause different pathologies of the urogenital system, including DDS. WT1 is composed of ten exons and encodes a protein with four zinc-finger motifs and transcriptional and tumour-suppressor activities. Alternative splicing generates four isoforms: the fifth exon may or may not be present, and an alternative splice site in intron 9 allows the addition of three amino acids (KTS) between the third and fourth zinc fingers of WT1 (ref. 17). Here we demonstrate that FS is caused by mutations in the donor splice site in intron 9 of WT1, with the predicted loss of the +KTS isoform. Examination of WT1 transcripts indeed showed a diminution of the +KTS/-KTS isoform ratio in patients with FS. PMID- 9398853 TI - Genetic analysis of ozone-induced acute lung injury in sensitive and resistant strains of mice. AB - Epidemiological studies have found air pollution to be associated with excessive mortality, particularly death from respiratory and cardiovascular causes. Interpretation of these findings is controversial, however, because toxicological mechanisms controlling mortality are uncertain. Susceptibility to many air pollutants entails an oxidative stress response. Accordingly, the best characterized oxidant air pollutant is ozone, which causes direct oxidative damage of lung biomolecules. An underlying characteristic derived from clinical and epidemiological studies of healthy and asthmatic individuals of all ages is marked variability in the respiratory effects of ozone. This susceptibility difference among humans suggests that genetic determinants may control predisposition to the harmful effects of ozone. Mice also vary considerably in their response to ozone. Moreover, ozone-induced differences in strain responses indicate that susceptibility in mice can be genetically determined. Therefore, we used inbred mice to investigate the genetic determinants of acute lung injury. Recombinant inbred (RI) strains derived from A/J (A) mice (sensitive) and C57BL/6J (B) mice (resistant) showed a continuous phenotypic pattern, suggesting a multigenic trait. Quantitative trait locus and RI analyses suggested three major loci linked to ozone susceptibility. Differences in phenotype ratios among the reciprocal back-crosses were consistent with parental imprinting. These findings implicate various genetic and epigenetic factors in individual susceptibility to air pollution. PMID- 9398855 TI - Genetic interaction between PARP and DNA-PK in V(D)J recombination and tumorigenesis. AB - Poly(ADP-ribose) polymerase (PARP) and DNA-dependent protein kinase (DNA-PK) are DNA break-activated molecules, Although mice that lack PARP display no gross phenotype and normal DNA excision repair, they exhibit high levels of sister chromatid exchange, indicative of elevated recombination rates. Mutation of the gene for DNA-PK catalytic subunit (Prkdc) cases defective antigen receptor V(D)J recombination and arrests B- and T-lymphocyte development in severe combined immune-deficiency (SCID) mice. SCID V(D)J recombination can be partly rescued in T-lymphocytes by either DNA-damaging agents (gamma-irradiation and bieomycin) or a null mutation of the p53 gene, possibly because of transiently elevated DNA repair activity in response to DNA damage or to delayed apoptosis in the absence of p53. To determine whether the increased chromosomal recombination observed in PARP-deficient cells affects SCID V(D)J recombination, we generated mice lacking both PARP and DNA-PK. Here, we show that thymocytes of SCID mice express both CD4 and CD8 co-receptors, bypassing the SCID block. Double-mutant T-cells in the periphery express TCR beta, which is attributable to productive TCR beta joints. Double-mutant mice develop a high frequency of T-cell lymphoma. These results demonstrate that increased recombination activity after the loss of PARP anti recombinogenic function can rescue V(D)J recombination in SCID mice and indicate that PARP and DNA-PK cooperate to minimize genomic damage caused by DNA strand breaks. PMID- 9398854 TI - Linkage analysis of susceptibility to ozone-induced lung inflammation in inbred mice. AB - Exposures to the common air pollutant ozone (O3) cause decrements in pulmonary function and induce airway inflammation that is characterized by infiltration of polymorphonuclear neutrophils (PMNs; refs 1-4). Because of the impact that O3 may have on public health, it is critical to identify susceptibility factors. Highly reproducible, significant inter-individual variations in human pulmonary function responses to O3 support the hypothesis that genetic background is an important determinant. Initial analysis of PMN responses to O3 exposure in segregant populations derived from inflammation-prone (susceptible) C57BL/6J (B6) and inflammation-resistant C3H/HeJ (C3) inbred mice indicated that susceptibility was controlled by a locus we termed Inf2 (ref. 7). Subsequent analyses with recombinant inbred strains suggested that a more complex interaction of genes is involved. In this report, we identify a quantitative trait locus (QTL) for O3 susceptibility on chromosome 17. Candidate genes for the locus include Tnf, the gene encoding the pro-inflammatory cytokine tumour necrosis factor-alpha (Tnf). Antibody neutralization of the protein product of this putative candidate gene significantly protected against O3 injury in susceptible mice. These results strongly support linkage of O3 susceptibility to a QTL on chromosome 17 and Tnf as a candidate gene. PMID- 9398856 TI - DNA-PKcs: a T-cell tumour suppressor encoded at the mouse scid locus. AB - Severe combined immunodeficiency (SCID) mice are defective in their ability to rearrange their variable (V), diversity (D) and joining (J) genetic elements to generate functional immunoglobulin (Ig) and T-cell receptor (TCR) molecules; as a result, they lack mature B and T cells. These mice are highly sensitive to ionizing radiation, suggesting that the product of the scid gene plays a critical role in both V(D)J recombination and DNA double-strand break repair. Recent studies suggest that the SCID defect lies in the gene encoding the catalytic subunit of DNA-dependent protein kinase (DNA-PK; refs 6-8), a nuclear protein made up of the Ku 70 and Ku 86 subunits as well as the large catalytic subunit, DNA-PKcs. Other reports have implied that the SCID phenotype correlates with nonsense mutations at the extreme 3' end of Prkdc, the DNA-PKcs gene. The identity of the gene remains in doubt, however, because the consequences of genetic inactivation of Prkdc have not been determined. This study shows that complete inactivation of Prkdc in a novel insertional mouse mutant recapitulates the SCID phenotype and that Prkdc and scid are alleic. Significantly, DNA-PKcs null mice demonstrate complete penetrance of thymic lymphoblastic lymphomas, strongly suggesting that Prkdc functions in mice as a T-cell tumour suppressor and, by virtue of its association with DNA repair and recombination, belongs to the 'caretaker' class of tumour-suppressor genes that includes ATM, BRCA1 and BRCA2 (ref. 15). PMID- 9398857 TI - The Fanconi anaemia proteins, FAA and FAC, interact to form a nuclear complex. AB - Fanconi anaemia (FA) is an autosomal-recessive disorder characterized by genomic instability, developmental defects, DNA crosslinking agent hypersensitivity and cancer susceptibility. Somatic-cell hybrid studies have revealed five FA complementation groups (A-E; refs 4-6) displaying similar phenotypes, suggesting that FA genes are functionally related. The two cloned FA genes, FAA and FAC, encode proteins that are unrelated to each other or to other proteins in GenBank. In the current study, we demonstrate the FAA and FAC bind each other and form a complex. Protein binding correlates with the functional activity of FAA and FAC, as patient-derived mutant FAC (L554P) fails to bind FAA. Although unbound FAA and FAC localize predominantly to the cytoplasm, the FAA-FAC complex is found in similar abundance in both cytoplasm and nucleus. Our results confirm the interrelatedness of the FA genes in a pathway, suggesting the cooperation of FAA and FAC in a nuclear function. PMID- 9398858 TI - Missense mutation in flavin-containing mono-oxygenase 3 gene, FMO3, underlies fish-odour syndrome. AB - Individuals with primary trimethylaminuria exhibit a body odour reminiscent of rotting fish, due to excessive excretion of trimethylamine (TMA; refs 1-3). The disorder, colloquially known as fish-odour syndrome, is inherited recessively as a defect in hepatic N-oxidation of dietary-derived TMA and cannot be considered benign, as sufferers may display a variety of psychosocial reactions, ranging from social isolation of clinical depression and attempted suicide. TMA oxidation is catalyzed by flavin-containing mono-oxygenase (FMO; refs 7,8), and tissue localization and functional studies have established FMO3 as the form most likely to be defective in fish-odour syndrome. Direct sequencing of the coding exons of FMO3 amplified from a patient with fish-odour syndrome identified two missense mutations. Although one of these represented a common polymorphism, the other, a C-->T transition in exon 4, was found only in an affected pedigree, in which it segregated with the disorder. The latter mutation predicts a proline-->leucine substitution at residue 153 and abolishes FMO3 catalytic activity. Our results indicate that defects in FMO3 underlie fish-odour syndrome and that the Pro 153- >Leu 153 mutation described here is a cause of this distressing condition. PMID- 9398860 TI - Reconstitution of human telomerase with the template RNA component hTR and the catalytic protein subunit hTRT. AB - The maintenance of chromosome termini, or telomeres, requires the action of the enzyme telomerase, as conventional DNA polymerases cannot fully replicate the ends of linear molecules. Telomerase is expressed and telomere length is maintained in human germ cells and the great majority of primary human tumours. However, telomerase is not detectable in most normal somatic cells; this corresponds to the gradual telomere loss observed with each cell division. It has been proposed that telomere erosion eventually signals entry into senescence or cell crisis and that activation of telomerase is usually required for immortal cell proliferation. In addition to the human telomerase RNA component (hTR; ref. 11), TR1/TLP1 (refs 12, 13), a protein that is homologous to the p80 protein associated with the Tetrahymena enzyme, has been identified in humans. More recently, the human telomerase reverse transcriptase (hTRT; refs 15, 16), which is homologous to the reverse transcriptase (RT)-like proteins associated with the Euplotes aediculatus (Ea_p123), Saccharomyces cerevisiae (Est2p) and Schizosaccharomyces pombe (5pTrt1) telomerases, has been reported to be a telomerase protein subunit. A catalytic function has been demonstrated for Est2p in the RT-like class but not for p80 or its homologues. We now report that in vitro transcription and translation of hTRT when co-synthesized or mixed with hTR reconstitutes telomerase activity that exhibits enzymatic properties like those of the native enzyme. Single amino-acid changes in conserved telomerase-specific and RT motifs reduce or abolish activity, providing direct evidence that hTRT is the catalytic protein component of telomerase. Normal human diploid cells transiently expressing hTRT possessed telomerase activity, demonstrating that hTRT is the limiting component necessary for restoration of telomerase activity in these cells. The ability to reconstitute telomerase permits further analysis of its biochemical and biological roles in cell aging and carcinogenesis. PMID- 9398861 TI - A case of acute lymphoblastic leukemia, near-triploidy, and poor outcome: characterization by fluorescence in situ hybridization using chromosome-specific libraries from all human chromosomes. AB - We have applied fluorescence in situ hybridization (FISH) using chromosome specific libraries from all 24 chromosomes on metaphase spreads from bone marrow cells, in order to resolve the chromosomal changes in leukemic cells from a 10 year-old boy with acute lymphoblastic leukemia (ALL), near-triploidy, and a subsequent poor outcome. The FISH analysis revealed a pattern of chromosome gains and losses that differed from all cases previously described. Most of the affected chromosomes were present in three copies (trisomy for chromosomes 1, 2, 5, 6, 7, 11, 12, 13, 14, 16, 17, 18, 19, 20, and 22), but the patient had four copies of chromosomes 8 and 21, two copies of chromosomes 3, 4, 9, 10, 15, and X, and one Y chromosome. No structural abnormalities could be detected. Thus, the karyotype of the malignant clone was 66,XXY-3,-4,+8,-9,-10,-15,+21. PMID- 9398859 TI - EWS/FLI1-induced manic fringe renders NIH 3T3 cells tumorigenic. AB - EWS/FLI1, a fusion gene found in Ewing's sarcoma, encodes a transcriptional regulator and promotes cellular transformation by modulating the transcription of specific target genes. We have found that EWS/FLI1 and structurally related fusion proteins upregulate manic fringe (MFNG), a recently described member of the Fringe gene family instrumental in somatic development. MFNG is also expressed in human tumour-derived cell lines expressing EWS/FLI1. Overexpression of MFNG in NIH 3T3 cells renders them tumorigenic in mice with severe combined immunodeficiency disease (SCID). These data demonstrate that part of the oncogenic effect of EWS/FLI1 is to transcriptionally deregulate a member of a family of morphogenic genes. PMID- 9398862 TI - Translocations between the long arms of chromosomes 1 and 5 in hematologic malignancies are strongly associated with neoplasms of the myeloid lineages. AB - The clinical, morphologic, and cytogenetic features of two hematologic malignancies--one acute myeloid leukemia with minimal differentiation (AML-MO) and one therapy-related myelodysplastic syndrome (MDS)--with unbalanced translocations between 1q and 5q are reported. The translocations resulted in loss of 5q material in both cases and gain of 1q in the MDS. A compilation of previously published hematologic malignancies with translocations involving the long arms of chromosomes 1 and 5 revealed a total of 23 cases--11 with unbalanced and 12 with balanced t(1;5)--with the following morphologies: 11 AML, three MDS, two Philadelphia-positive chronic myeloid leukemias, three chronic myeloproliferative disorders, three acute lymphoblastic leukemias, and one chronic lymphocytic leukemia. Four patients had received chemotherapy including alkylating agents for a previous malignancy and one had been exposed to thorotrast. Among the 14 patients for whom survival data exist, all except three have died. The t(1;5) was found as the sole abnormality in six cases, whereas it was apparently secondary--occurring in a subclone or together with the well-known primary abnormalities t(8;21), t(9;22), and t(15;17)--in nine cases. The breakpoints in 1q varied from 1q11 to 1q43, with a clustering to 1q21-23, and the 5q breaks occurred in 5q11 to 5q35, mainly in the distal 5q3 region. The unbalanced 1;5 translocations resulted in gain of 1q material in eight of the 11 cases, 1q21-1qter being duplicated in four of them, and in loss of 5q, most often the 5q3 region, in 10 of the neoplasms. We conclude that translocations between 1q and 5q, although cytogenetically heterogeneous, are associated with hematologic malignancies of the myeloid lineages and with previous mutagenic exposure, and that t(1;5) seems to confer a poor prognosis. PMID- 9398864 TI - Specific cytogenetic aberrations in two novel human prostatic cell lines immortalized by human papillomavirus type 18 DNA. AB - Using chromosome banding and fluorescence in situ hybridization (FISH) with painting probes, sequential cytogenetic analysis was performed of two novel prostate cell lines, PZ-HPV-7 and CA-HPV-10, established by human papillomavirus (HPV) 18 DNA transformation. PZ-HPV-7 originates from a normal diploid prostate epithelial cell strain. PZ-HPV-7 progressed from an initial diploid to a hypertetraploid chromosome number with a relative gain of chromosomes 5 and 20 (7 to 8 copies each). Structural changes were limited; 3p- (2 copies), 3q- (1 copy), and possibly a der(16p;12q). CA-HPV-10 originates from an epithelial cell strain derived from a high-grade human prostate cancer specimen, which showed several karyotypic abnormalities including an extra Y chromosome and double minutes (dmin). In early passage the karyotype of CA-HPV-10 appeared unstable with a decreasing number of cells exhibiting dmin. In late passage the dmin were replaced by a large homogeneously staining region (hsr) on 9p+ marker. The hsr was shown by FISH to be of chromosome 1 origin. The modal number was mainly hypertriploid (72, range 69 to 75). Loss of Y was remarkable (0 to 1 copy). Consistent markers included two copies each of del(1)(q12q31) and der(9)t(1;9)(?;p22), and one der(11)t(4;11) (?;q21). HPV type 18 genomic integration sites were identified on 1p for PZ-HPV-7 and on the 9p+ marker for CA HPV-10. In conclusion, both PZ-HPV-7 and CA-HPV-10 showed clonal cytogenetic changes. These two cell lines constitute a novel in vitro model to study the mechanisms involved in human prostate carcino-genesis. PMID- 9398863 TI - Interphase detection of BCL6/IgH fusion gene in non-Hodgkin lymphoma by fluorescence in situ hybridization. AB - We characterized a t(3;14)(q27;q32) translocation in nine patients with B-cell, non-Hodgkin lymphoma (B-NHL) by fluorescence in situ hybridization (FISH). Fluorescence in situ hybridization with immunoglobulin heavy chain (IgH) and BCL6 gene probes detected t(3;14) rapidly and accurately, including complex t(3;14) in three patients; one with t(3;12;8;14)(q27;p13;q24.1;q32) and two with t(3;?;14)(q27;?;q32). Among these nine patients, seven escaped from cytogenetic detection by our G-banding analysis. Double-color FISH with IgH (Y6) and BCL6 (cosB5-1) showed fusion of BCL6 and IgH genes on der(3)t(3;14) in all nine patients, suggesting that der(3) may play a critical role in the development of lymphoma carrying complex as well as standard t(3;14) translocations. BCL6/IgH fusion gene was also demonstrated in interphase nuclei at a frequency of 23% to 91.5% over the cut-off value in control studies (9.0 +/- 2.76%). The breakpoints assessed by FISH with two cosmid clones containing BCL6 probes, cosB5-1 and cosB5 2, were within the cluster region in seven patients including one with complex type, but were not evaluated in two patients with t(3;?;14), because of the loss of partner chromosome. Using double-color FISH with these two BCL6-specific probes, none of an additional 32 patients in whom mitotic spreads were available showed 3q27 translocations. Fluorescence in situ hybridization with IgH and BCL6 gene probes is a rapid and sensitive method to detect t(3;14) in routine cytogenetic studies. PMID- 9398865 TI - Highly complex chromosomal aberrations in bone marrow of a patient with metastatic prostate neoplasm. AB - Prostate cancer is the single most common malignancy among men in North America. Nevertheless, cytogenetic evaluation of bone marrow in patients with metastatic prostate neoplasm has been rare and, to date, only five such patients have been reported. We report an additional case where chromosomal abnormalities of a bizarre nature were found in the bone marrow. Though cytogenetic findings in prostate cancer are heterogeneously complex, the chromosome regions involved include 1p, 1q, 7q, 8p, 10q, 12p, and 17q and are considered hot spots. What is the significance of these so-called hot spots in metastasis of prostatic cancer to the bone marrow? At present, no meaningful conclusion can be drawn, as data are limited, but accumulation of such cases may provide valuable information concerning the role of chromosomal abnormalities in patients--specifically with metastatic stage--and may help urologists during therapeutic decision making, particularly if a genetic marker for aggressiveness can be determined. PMID- 9398866 TI - Cytogenetic findings in malignant mixed mesodermal tumors of the uterus. AB - Cytogenetic analyses of four malignant mixed mesodermal tumors (MMMT) of the uterus are reported, of which one was of the homologous type and three of the heterologous. Karyotypic analyses were obtained in two cases from original tumors and in two cases from tumors xenotransplanted into nude mice. The karyotype of the homologous MMMT was normal in three different passages of a nude mice xenograft line established from the primary tumor. The heterologous tumors showed normal karyotype in one case and hyperdiploid and near triploid range with extensive numerical and structural rearrangements in two cases. Deletion of chromosome 1 at p32, and deletion of chromosome 11 at q13 were common markers in anomalous cases. The chromosomes most often involved in structural rearrangements were chromosomes 1, 9, 11, 12, 17, and 19. Double minutes, homogeneously staining regions, and telomeric association were also seen. PMID- 9398867 TI - Increased levels of mutagen-induced chromosome breakage in Down syndrome children with malignancy. AB - Even though an association between Down syndrome (DS) and malignancies has been established, the mechanism behind this is still unclear. We therefore investigated constitutional chromosomal abnormalities and bleomycin-induced chromosome sensitivity in 12 DS children, 8 DS children with malignancies, and 10 normal controls to explore whether these factors play any role in cancer predisposition. Trisomy 21 was the only constitutional cytogenetic abnormality observed in all the DS children. But there was significant variation between the patients and controls with regard to bleomycin sensitivity. Compared to the normal controls, all the DS patients expressed significantly higher chromosomal breaks per cell (b/c) values indicating sensitivity to bleomycin. Furthermore, DS children with malignancies demonstrated significantly higher b/c values than DS children with malignancies. These results permit us to assume that DS children showing mutagen hypersensitivity may be having defective DNA repair competence and hence may be predisposed to malignancies. PMID- 9398868 TI - Monosomy 22 in a fibrothecoma. AB - We found a 45,XX, -22 karyotype as the sole chromosome change in a fibrothecoma of a 67-year-old woman. These cytogenetic findings are discussed in the light of relevant cytogenetic and pathology literature data on ovarian sex cord tumors. PMID- 9398869 TI - Chromosome analysis in angiomyolipoma. AB - Cytogenetic data on angiomyolipomas are scarce. We report a series of 10 cases from nine patients without clinical history of tuberous sclerosis. We found chromosome abnormalities in two, and in both cases the chromosomal changes were only numerical ones, i.e., trisomy 7 and trisomy 8. PMID- 9398870 TI - Immunohistochemical detection of Helicobacter pylori in the surface mucous gel layer and its clinicopathological significance. AB - BACKGROUND: Attempts have been made to develop an accurate method for detecting Helicobacter pylori in histological sections. MATERIALS AND METHODS: Biopsy specimens were obtained from the stomachs of 167 patients with gastric ulcer (33), duodenal ulcer (52), gastroduodenal ulcer (15), chronic gastritis (45), and normal mucosa (22) before antimicrobial treatment and from 108 of these patients after treatment. Biopsy specimens were (1) cultured, (2) fixed in 10% buffered formalin, or (3) fixed in Carnoy's solution. The latter method was employed to preserve the surface mucous gel layer (SMGL) covering gastric surface mucous cells. Histological sections were stained with hematoxylin and eosin (H&E), with immunostaining using a commercially available polyclonal anti-H. pylori antibody. RESULTS: Cultures were positive for H. pylori in 61% of the cases before treatment and in 16% after treatment; by H&E staining using formalin-fixed materials: 70% and 9%; by immunostaining using formalin-fixed materials: 78% and 21%; and by immunostaining using Carnoy-fixed materials: 85% and 41% of biopsy specimens, respectively. The difference in detection rates between materials fixed in formalin and those in Carnoy's solution was due to the detection of H. pylori in the SMGL by the latter, especially after antimicrobial treatment. CONCLUSIONS: Immunostaining for H. pylori using materials fixed in Carnoy's solution revealed H. pylori in the SMGL as well as on the surface mucous cells and in the gastric pits and permitted the optimal detection of H. pylori in tissue sections. PMID- 9398872 TI - Serum and gastric luminal epidermal growth factor in Helicobacter pylori associated gastritis and peptic ulcer disease. AB - BACKGROUND: Helicobacter pylori is the cause of chronic (type B) gastritis, duodenal ulceration (DU), and gastric ulceration (GU). Smoking is associated with delayed ulcer healing. Epidermal growth factor (EGF) is produced in the salivary and Brunner's glands of the upper gastrointestinal tract, inhibits gastric acid secretion, and is a powerful mitogen. MATERIALS AND METHODS: We sought to determine gastric luminal EGF (GL-EGF) in smokers and patients with Hp-associated DU and the effects of Hp eradication. Our aim was to determine GL-EGF in patients with GU and the effect of ulcer healing and to measure serum EGF in patients with Hp gastritis with or without DU disease. RESULTS: GL-EGF was reduced in smokers compared to controls (p = .008). Subjects with HP gastritis had reduced GL-EGF compared to controls (p = .0002). There was no difference in GL-EGF between Hp positive subjects who had DU and those with chronic gastritis alone. Eradication of Hp from those patients with DU had no effect on the low levels of GL-EGF. There was no difference between GL-EGF in Hp gastritis alone and in Hp-associated active GU. GL-EGF fell after ulcer healing (p = .04), a difference confirmed by analysis of paired samples from patients before and after ulcer healing (p = .03). There was no difference in serum EGF between controls and subjects with Hp infection. There was no difference in serum EGF in subjects with DU associated and non-ulcer-associated gastritis. CONCLUSIONS: Subjects with Hp gastritis, or those who smoke, had low concentrations of GL-EGF regardless of whether DU was present. Eradication of Hp did not return the concentrations of GL-EGF to normal in DU subjects. Individuals and Hp gastritis and inactive GU had low levels of GL EGF compared to non-ulcer Hp infection. The relative increase in GL-EGF that occurred with ulceration of the gastric mucosa may have resulted from the development of an ulcer-associated cell lineage. Serum EGF did not play a role in the pathogenesis of Hp gastritis or of associated DU ulcer disease. PMID- 9398871 TI - Helicobacter pylori and the surface mucous gel layer of the human stomach. AB - BACKGROUND: The colonization of Helicobacter pylori in the surface mucous gel layer (SMGL) was investigated. MATERIALS AND METHODS: Surgically removed stomachs were obtained from patients and included gastric ulcer (4 cases), duodenal ulcer (2), and gastric cancer (24). Five of these cases were examined at 8, 19, 28, 143, and 171 days after the end of eradication therapy. For the preservation of the SMGL, these specimens were fixed in cold Carnoy's solution, cleared in xylene, and embedded in paraffin. Serial sections were obtained and were stained by dual staining with the galactose oxidase-cold thionin Schiff reaction followed by paradoxical Concanavalin A staining and immunostaining for H. pylori. RESULTS: H. pylori characteristically attached to surface mucous cells and colonized in the SMGL H. pylori in the SMGL was more abundant than that attached to the surface mucous cells. The degree of H. pylori infection both on the surface of surface mucous cells and in the SMGL correlated well with the severity of gastritis. In the SMGL, this organism obviously preferred to colonize in the layer of surface mucous cell-type mucins, and the multilaminated structure of the SMGL deteriorated markedly. Eradication of H. pylori restored the structure of the SMGL, and the inflammatory reaction decreased gradually. CONCLUSION: The SMGL is an indispensable site of H. pylori colonization, and this organism damaged the gastric mucosa partially by causing deterioration of the SMGL. Removal of the organism from the SMGL should be considered for eradication of this organism. PMID- 9398873 TI - The correlation in dyspeptic patients of Helicobacter pylori infection with changes in interdigestive gastroduodenal motility patterns but not in gastric emptying. AB - BACKGROUND: Available data conflict regarding the possible relation between chronic gastritis, Helicobacter pylori (Hp), and gastric motor disorders in nonulcer dyspepsia. The aim of this study, therefore, was (1) to evaluate both gastroduodenal fasting motility and gastric emptying in subjects with functional dyspepsia, with and without gastritis, and (2) to correlate the motility pattern to Hp infection. MATERIALS AND METHODS: Thirty-eight patients were studied, 20 positive for Hp infection (15 with gastritis) and 18 Hp-negative (8 with gastritis). All the subjects underwent 240-minute manometric recording of the interdigestive migrating motor complex, with evaluation of gastric and duodenal motility pattern and scintigraphic study of gastric emptying. RESULTS: Whereas gastric emptying half-time did not differ in the subgroups of dyspeptic patients, a significant reduction of gastric phase IIIs of the migrating motor complex was detected between Hp-positive and Hp-negative subjects, both in overall patients (p < .01) and in patients with gastritis (p < .05). CONCLUSION: Hp infection seems to be related to a reduction of interdigestive gastric activity fronts, though it does not affect gastric emptying. The conflicting data regarding gastric emptying and interdigestive motility in Hp infection could be explained as probably investigating two different functional aspects. PMID- 9398874 TI - Triple therapy with lansoprazole, clarithromycin, and amoxicillin for the cure of Helicobacter pylori infection: a short report. AB - BACKGROUND: Given the therapeutic potential of proton pump inhibitor-based triple therapy for successful cure of Helicobacter pylori infection, we evaluated the efficacy and safety of lansoprazole with clarithromycin and amoxicillin in an open-label, single-center study. MATERIALS AND METHODS: H. pylori-positive patients self-administered lansoprazole, 30 mg; clarithromycin, 500 mg; and amoxicillin, 1 gm bid for 14 days. Patients were assessed pretreatment, at which time the presence of H. pylori was documented by rapid urease test, culture, or histology, following study drug administration (week 2) for a brief evaluation only, and at least 4 weeks posttreatment (week 6), which included endoscopy with collection of biopsy specimens for culture and histology testing. RESULTS: Primary clarithromycin and metronidazole resistance were observed in 6% (2 of 30) and 43% (13 of 30) of study patients, respectively. One month after the end of therapy, H. pylori infection was cured in 23 of 25 patients (92%; 95% confidence interval, 74%-99%). The triple-therapy regimen was well-tolerated; 17% of patients (5 of 30) reported mild to moderate adverse effects during the treatment period. CONCLUSION: A 2-week, triple-drug combination of lansoprazole, clarithoromycin, and amoxicillin is highly effective for cure of H. pylori infection. Additionally, the triple-drug combination was well-tolerated by patients infected with H. pylori. PMID- 9398876 TI - Seven-day triple therapy with lansoprazole, clarithromycin, and metronidazole for the cure of Helicobacter pylori infection: a short report. AB - BACKGROUND: To refine our understanding of anti-Helicobacter pylori treatment regimens further, we evaluated the efficacy and safety of lansoprazole given in combination with clarithromycin and metronidazole for 7 days in an open-label, multicenter study. MATERIALS AND METHODS: H. pylori-positive patients self administered lansoprazole, 30 mg; clarithromycin, 500 mg; and metronidazole, 500 mg bid for 7 days. Patients were assessed at pretreatment, at which time the presence of H. pylori was documented by rapid urease test or histology and culture, following study drug administration (week 1) for a brief evaluation only, and at least 4 weeks posttreatment (week 5), including endoscopy with collection of biopsy specimens for culture and histology testing. RESULTS: Of the 60 patients enrolled in the study, 59 had confirmed H. pylori infection, and 51 were included in an intent-to-treat analysis of efficacy. Primary metronidazole and clarithromycin resistance were observed in 84% and 8% of study patients, respectively. One month after the end of therapy, H. pylori infection was cured in 40 of 51 patients (78%); 95% confidence interval, (65%-89%). The triple therapy regimen was well-tolerated, with only 2 patients (4%) requiring premature withdrawal from the study due to treatment-related adverse events. Taste perversion (15.0%) and diarrhea (11.7%) were the most frequently reported adverse events possibly or probably related to study medication during the treatment period. CONCLUSION: Despite a high prevalence of metronidazole resistance, a 1 week, triple-drug combination of lansoprazole, clarithromycin, and metronidazole is effective treatment for and well-tolerated by patients with H. pylori infection. PMID- 9398875 TI - Double-blind, multicenter evaluation of lansoprazole and amoxicillin dual therapy for the cure of Helicobacter pylori infection. AB - BACKGROUND: Treatment with amoxicillin plus omeprazole results in disappointing cure rates of Helicobacter pylori infection. The minimal inhibitory concentration of lansoprazole for H. pylori in vitro is lower than that for omeprazole, prompting interest in treatment with amoxicillin plus lansoprazole. MATERIALS AND METHODS: H. pylori-infected patients with endoscopically documented duodenal ulcer either currently or within the past year were randomized to 14 days of (1) lansoprazole, 30 mg bid, plus amoxicillin, 1 gm tid; (2) lansoprazole, 30 mg tid, plus amoxicillin, 1 gm tid; (3) lansoprazole, 30 mg tid alone; or (4) amoxicillin, 1 gm tid alone. Endoscopy was done at enrollment and at 4 to 6 weeks after completion of treatment or for recurrent symptoms. H. pylori status was assessed by culture and histology. Ulcer prevalence was evaluated at follow-up endoscopy. RESULTS: Two hundred sixty-two patients met enrollment criteria and were treated. By per-protocol analysis, H. pylori infection was cured in 57% of those treated with lansoprazole twice daily plus amoxicillin and in 67% of those treated with lansoprazole three times daily plus amoxicillin, compared with 0% treated with lansoprazole alone or amoxicillin alone (p < .001 for dual therapy versus either monotherapy). Amoxicillin resistance was not observed. At follow-up endoscopy, ulcer prevalence was 17% in patients treated with lansoprazole twice daily plus amoxicillin, 23% in those treated with lansoprazole three times daily plus amoxicillin, 33% in those treated with lansoprazole alone, and 35% in those treated with amoxicillin alone (p = .024; lansoprazole twice daily plus amoxicillin versus amoxicillin alone). CONCLUSIONS: Treatment with amoxicillin plus lansoprazole, 30 mg tid, led to cure of H. pylori infection in 67% of patients with active or recently healed duodenal ulcer. PMID- 9398877 TI - The best gastric site for obtaining a positive rapid ureas test. AB - BACKGROUND: Rapid ureas tests (RUTs) provide a simple, sensitive method of detecting Helicobacter pylori infection. OBJECTIVES: Our aim, therefore, was to determine whether the yield of detecting H. pylori infection by RUT varied depending on the site of gastric biopsy. MATERIALS AND METHODS: Gastric biopsies were obtained from 50 patients for RUT by use of hpfast (GI Supply, Camp Hill, PA). Biopsies were taken from the prepyloric greater curve antrum, from the gastric angle, and from the greater curve in mid-corpus. One biopsy specimen was placed in the RUT gel, and the biopsy from the adjacent mucosa was placed in formalin for subsequent histological evaluation by using the Genta stain. RUTs were examined and scored at intervals of 5, 10, 15, 30, and 45 minutes and after 1, 2, 4, and 24 hours. RESULTS: Fifty patients were entered in the rest (150 RUTs), 32 having H. pylori infection. There were no false-positive RUTs (specificity, 100%). The gastric angle site was positive in 100%. The prepyloric site was positive in 87%, and the corpus site was positive in 84.4% (p < .052 for angle or prepyloric antrum versus corpus). The most common pattern was for all to be positive (74%). The median time to positivity was similar with angle and prepyloric sites (37.5 and 60 minutes, respectively, p = not significant) and shorter than the corpus biopsy (180 minutes); (p < .05 for angle or prepyloric antrum versus corpus). CONCLUSION: The maximum probability for detecting H. pylori infection using a RUT is to obtain a biopsy from the gastric angle. To prevent missing a positive result when intestinal metaplasia is present, we recommend that (at a minimum) biopsies be taken from both the angle and the corpus. PMID- 9398878 TI - Duodenal ulcer healing after 7-day treatment: a pilot study with lansoprazole, amoxicillin, and clarithromycin. PMID- 9398879 TI - How to manage the dyspeptic patient. PMID- 9398880 TI - Comparison of one or two weeks of lansoprazole, amoxicillin, and clarithromycin in the treatment of Helicobacter pylori. AB - BACKGROUND: The simplest, most effective, and least expensive Helicobacter pylori therapy remains to be determined. Two weeks of 30 mg lansoprazole bid, 1 gm amoxicillin bid, and 500 mg clarithromycin bid (LAC2) had been shown to be an effective therapy for H. pylori. The aim of this study was to assess whether 1 week of this regimen (LAC1) would have a similar efficacy. MATERIALS AND METHODS: H. pylori-positive patients assessed histologically, by rapid urease test, microbiologically, and by a 13C-urea breath test (13C-UBT) were randomized to receive either LAC1 or LAC2 in a single-center open study. Patients were interviewed 1 to 3 days after completion of therapy to evaluate adverse events and compliance. Efficacy was determined by 13C-UBT at least 4 weeks after antibiotic therapy. RESULTS: Seventy evaluable patients were randomized to receive LAC1 (n = 33) LAC2 (n = 37). Of the 33 LAC1 patients, 30 (91%) were treated successfully (95% confidence interval (CI) = 76-98%), compared with 32 of 37 (86%) in the LAC2 group (95% CI = 71-96%). There was no difference in efficacy between the two groups (Fisher's exact test p = 1.0; 95% CI = -10.3%-19.2%). Patients taking LAC1 experienced significantly fewer severe adverse events than those taking LAC2 (Mann-Whitney U test). One of 64 patients had primary resistance to clarithromycin, and treatment was unsuccessful in this case. Six of the 7 remaining treatment failures developed secondary resistance to clarithromycin. CONCLUSIONS: LAC1 is as effective as LAC2 and is associated with less toxicity. Posttreatment clarithromycin resistance is common in patients who do not experience success with therapy. PMID- 9398881 TI - Epidemiology of gastric non-Hodgkin's lymphoma patients: parallels with Helicobacter pylori. AB - BACKGROUND: The incidence of gastric non-Hodgkin's lymphoma (NHL) is increasing in the United States. However, little is known about the etiology of the disease. Some studies have shown an association between gastric NHL and Helicobacter pylori. No study has specifically delineated demographic features that distinguish gastric NHL patients from nongastric NHL patients. MATERIALS AND METHODS: To obtain information about the differences between gastric and nongastric NHL patients, we conducted a hospital chart review study. We examined charts of all 25 cases of primary gastric NHL, as well as charts of 75 randomly selected nongastric NHL patients as controls. All patients were seen in the Division of Oncology at Stanford University Medical Center from 1972 to 1991. Demographic information was tabulated, and differences between the cases and controls were noted. The identified risk factors were determined by both univariate and logistic regression analyses. RESULTS: There was no difference between gastric NHL cases and nongastric controls with respect to age, gender, race, and family history of any cancer. However, in logistical regression, persons with gastric NHL were more likely than those with other forms of NHL to be born outside the United States (odds ratio = 12.8; 95% confidence interval = 2.9-56.0) and also to have a family history of stomach cancer (odds ratio = 18.4; 95% confidence interval 2.1-160.1). CONCLUSIONS: Gastric NHL is more likely than NHL at other sites to occur in persons with a family history of gastric cancer or in those born in developing countries. This epidemiological pattern supports the identified role of H. pylori in the development of gastric lymphoma. PMID- 9398882 TI - Is Helicobacter pylori transmitted from cats to humans? AB - BACKGROUND: Subsequent to the isolation of Helicobacter pylori from domestic cats, it has been suggested that the organism might be transmitted from cats to humans. This hypothesis has already gained considerable media attention. MATERIALS AND METHODS: In a previous study of risk factors for H. pylori infection, 447 factory workers from Stoke on Trent in the UK had provided blood samples for H. pylori serological workup. They had also completed a detailed questionnaire concerning their living conditions, including the possession of any household pets, in childhood. Logistic regression was used to assess the association between cat ownership in childhood and H. pylori seropositivity. RESULTS: After adjustment for potential confounders, it was found that subjects who had owned a pet as a child were slightly more likely to be H. pylori seropositive than subjects who had not. There was, however, no difference between subjects who had owned a cat and those with other pets. CONCLUSIONS: These data do not support the hypothesis that H. pylori infection might be transmitted from cats to humans. PMID- 9398883 TI - Helicobacter pylori and socioeconomic factors in Russia. AB - BACKGROUND: The factors influencing the acquisition and prevalence of Helicobacter pylori infection remain incompletely understood. In Russia, the demographic and socioeconomic factors are relatively similar, allowing investigation of risk factors that might not be identifiable in a more diverse population. MATERIALS AND METHODS: Sero-prevalence of H. pylori infection was studied in 520 asymptomatic individuals between the ages of 1 and 75 years, residing in St. Petersburg, Russia. Forty-four children lived in orphanages or communal apartments. Demographic information and socioeconomic factors were evaluated, including educational level, income, and living conditions. Helicobacter pylori status was evaluated by using an enzyme-linked immunosorbent assay for anti-H. pylori IgG. RESULTS: The prevalence of H. pylori infection was 44% in children and 88% in adults (P < .001). In adults, H. pylori prevalence was independent of socioeconomic factors. The crude and the age-adjusted odds ratios (ORs) in children showed an inverse correlation between the mother's educational level and H. pylori seropositivity [e.g., OR, 1.8; (95% confidence interval (CI) = 1-3.2] for children whose mothers completed only 8 to 10 years of school compared to children whose mothers completed university. Overcrowding in childhood also was associated with increased H. pylori prevalence. Children from orphanages and communal apartments had the highest crowding index and also were at the greatest risk for H. pylori acquisition (age-adjusted OR, 2.1; 95% CI = 1.2-2.5). CONCLUSIONS: The prevalence of H. pylori infection in Russia correlated with socioeconomic factors, suggesting there are differences sufficient to affect H. pylori transmission. The prevalence of H. pylori infection during childhood forms the basis for the variances in prevalence among populations. PMID- 9398884 TI - Is antrum or corpus the best site for culture of Helicobacter pylori? AB - BACKGROUND: Isolating Helicobacter pylori on culture media and performing antibiotic susceptibility testing is potentially the most useful tool for guiding antibiotic therapy, especially when antimicrobial resistance is suspected. The aim of this study was to determine whether the yield of H. pylori culture was related to the site from which the gastric specimen was obtained either before or after therapy. METHODS: Gastric mucosal biopsies from the antrum and the corpus of the stomach were cultured. H. pylori status was determined by histological assessment using the Genta stain. RESULTS: Fifty-two patients with documented H. pylori infection were studied: Twenty-three were tested before antibiotic therapy and 29 after therapy had failed. In 47 patients (90%), both antral and corpus culture specimens were positive. In 5 patients (10%), only one site was positive, with three false-negative antral and two false negative corpus cultures. The overall sensitivity of culture in detecting H. pylori infection was 95% (95% confidence interval = 89-98%) and was not significantly different for the antrum or corpus, either before or after therapy. CONCLUSION: Culture of gastric biopsies from either the antrum or the corpus has an excellent diagnostic yield even in patients who failed antimicrobial therapy. PMID- 9398885 TI - Different expression of Helicobacter pylori gastritis in children: evidence for a specific pediatric disease? AB - BACKGROUND: Infection with Helicobacter pylori causes active chronic gastritis. Once the infection is acquired, gastritis will persist for almost the rest of one's life. To date, very few data are available on H. pylori gastritis in relation to age. Therefore, we attempted to investigate whether H. pylori gastritis in children exhibits features different from H. pylori gastritis in adults of two different age groups. MATERIALS AND METHODS: Fifty consecutive children with a median age of 11 years (range, 3-18 years) were compared with two groups of 50 adult patients, one group with a median age of 43 (range, 19-56 years) and another group with a median age of 70 years (range, 59-86 years). All patients had H. pylori gastritis unrelated to active peptic ulcer disease. Two biopsy specimens were taken from the antrum and two from the corpus, and the following gastritis parameters were evaluated: degree and activity of gastritis, H, pylori colonization, replacement of foveolar epithelium by regenerative epithelium, mucous depletion, presence of atrophic gastritis with intestinal metaplasia, and presence of lymphoid follicles. RESULTS: Degree and activity of gastritis, extent of H. pylori colonization, degree of replacement by regenerative epithelium, extent of mucous depletion, degree of atrophic gastritis with intestinal metaplasia, and the presence of lymphoid follicles in the antrum, as well as the presence of lymphoid follicles in the corpus differed significantly (chi-square test: p < .05). All these differences--except the once frequent occurrence of atrophic gastritis with intestinal metaplasia in adults- were attributable to a higher expression of these gastritis parameters in children. CONCLUSIONS: We conclude that H. pylori gastritis, particularly in the antrum, is more severely expressed in childhood. One reason for this might be a child-specific immune response to an infection with H. pylori. Alternatively, infection may represent a pediatric disease characterized by a nonatrophic, highly expressed form of gastritis, which changes its appearance once the host becomes adapted over time. PMID- 9398886 TI - Mixed infection with cagA-positive and cagA-negative strains of Helicobacter pylori. AB - BACKGROUND: Helicobacter pylori infection has been implicated strongly in the pathogenesis of gastritis, peptic ulcer disease, gastric adenocarcinoma, and gastric lymphoma, but the reasons for these widely different clinical outcomes are unknown. The aim of this study was to determine whether these differences could be due in part to mixed infection in the same individual, with bacteria having differences in pathogenic factors associated with ulcers. MATERIALS AND METHODS: The cagA gene of H. pylori was used to test for mixed infection because it is present in only some strains, and its presence has been associated with ulcers. Polymerase chain reaction (PCR) assays for the cagA gene were applied to H. pylori culture isolates and endoscopic gastric aspirates. Individual bacterial clones were tested for genetic similarity by random primer amplification and restriction endonuclease digestion of urease gene PCR products. RESULTS: The majority of H. pylori-positive patients had strongly cagA-positive culture isolates and endoscopic samples (62.5% and 69.6%, respectively). However, many of these patients had evidence of mixed infection with cagA negative and cagA positive strains in cultures isolates and endoscopic samples (25% and 17.4%, respectively). Mixed infection was found to be due to genetically unrelated strains in two patients in whom genetic analysis was performed. CONCLUSION: Mixed infection with differences in substrain pathogenic factors might occur in H. pylori infection and might contribute to differences in clinical outcome. PMID- 9398887 TI - Presence of the cagA gene in the majority of Helicobacter pylori strains is independent of whether the individual has duodenal ulcer or asymptomatic gastritis. AB - BACKGROUND: Helicobacter pylori infection presents as many different diseases, including asymptomatic gastritis, peptic ulcer disease, and gastric cancer. Although the virulence factor(s) responsible for different H. pylori-related diseases have not been identified, several candidate genes are being investigated for such an association. The polymerase chain reaction (PCR) frequently is used to assess the presence of genetic factors associated with pathogenesis of disease; the cagA gene and its product have been postulated to have a disease specific relationship to peptic ulcer and gastric cancer because of differential expression in these diseases compared to histological gastritis alone. MATERIALS AND METHODS: Genomic DNA was amplified by PCR, using synthetic oligonucleotide primers to the cagA gene to determine the prevalence of the cagA gene in 60 H. pylori isolates obtained from well-documented duodenal ulcer or asymptomatic gastritis patients (30 each). Results were confirmed by hybridization with a 1.4 Kb cagA probe. RESULTS: The expected PCR product was obtained in 90% of isolates from duodenal ulcer patients, compared to 70% of isolates from individuals with asymptomatic gastritis. The PCR products were polymorphic in size, suggesting cagA gene sequence differences among isolates. Evaluation for the presence of the cagA gene by hybridization with a 1.4-Kb cagA probe showed a homologous product in 29 of 30 strains [96.7%; 95% confidence interval (CI) = 83-100%] from duodenal ulcer patients versus 25 of 30 strains (83.3%; 95% CI = 65-94%) obtained from individuals with asymptomatic gastritis (p = 0.19). CONCLUSIONS: The high prevalence of the cagA gene in asymptomatic gastritis suggests that it will not prove to be a useful marker to distinguish more virulent or disease-specific H. pylori strains. The genetic heterogeneity among H. pylori strains makes PCR an unwise choice as the single method to determine prevalence of a putative virulence factor. In evaluation of the prevalence of a gene or genetic factor in a population of H. pylori, hybridization with extended probes might be important to ensure that the results are representative of the organism's genotype. PMID- 9398888 TI - Conservation and diversity of the Helicobacter pylori copper-transporting ATPase gene (copA) sequence among Helicobacter species and Campylobacter species detected by PCR and RFLP. AB - BACKGROUND: Helicobacter pylori is a causative pathogen of such human stomach diseases as chronic type B gastritis, ulcer, and possibly gastric carcinoma. As a cofactor in various redox enzymes and an essential trace metal required for the synthesis of metalloproteins, copper might play a role in the pathogenesis of H. pylori. A gene, copA, associated with copper transport, has been isolated from H. pylori UA802. In this study, conservation and diversity of this gene were analyzed among some Helicobacter and Campylobacter species. MATERIALS AND METHODS: Twenty-one clinical isolates and strains of helicobacters and campylobacters were used in this study. Methods including polymerase chain reaction (PCR) amplification, restriction fragment-length polymorphisms (RFLPs), and hybridization were employed to carry out this work. RESULTS: The copA gene was highly conserved in all the H. pylori isolates tested (Helicobacter nemestrinae and Helicobacter felis but not in Helicobacter mustelae and the Campylobacter species), whereas the sequence downstream of the copA appears to diverge among H. pylori isolates. In addition, two restriction patterns of the PCR-amplified copA fragments from seven H. pylori isolates and H. nemestrinae were identified, and the RFLP of H. nemestrinae was identical to that of one of the H. pylori isolate group. CONCLUSIONS: The adenosine triposphatase-derived copper-transporting mechanism is employed by various H. pylori strains, H. nemestrinae, H. felis, and perhaps by other Helicobacter species. The nucleotide mutations have risen in the copA gene. It appears that there is a genetic relatedness of the copA gene to H. pylori and H. nemestrinae. PMID- 9398889 TI - Treatment of Helicobacter pylori infection: summary of a meeting at the Fourth United European Gastroenterology Week, September 20, 1995. PMID- 9398890 TI - A standardized system of abbreviation for describing treatment regimens for Helicobacter pylori. PMID- 9398891 TI - Quadruple therapy: the golden bullet or a drug too far? PMID- 9398892 TI - Transformation of H. pylori by plasmid. PMID- 9398893 TI - Double-blind, multicenter, placebo-controlled evaluation of clarithromycin and omeprazole for Helicobacter pylori-associated duodenal ulcer. AB - BACKGROUND: Eradication of Helicobacter pylori leads to faster ulcer healing and a significant decrease in ulcer recurrence. Clarithromycin is the most effective monotherapy for eradicating H. pylori from the gastric mucosa, and omeprazole frequently is used for the treatment of duodenal ulcer disease, prompting the interest to investigate rigorously the combination of clarithromycin and omeprazole for eradicating H. pylori. MATERIALS AND METHODS: The aim of this double-blind, randomized, multicenter (n = 30), multinational (n = 10) study was to compare clarithromycin and omeprazole with omeprazole monotherapy for the eradication of H. pylori from the gastric mucosa, endoscopic healing, and reduction of symptoms and ulcer recurrence in patients with active duodenal ulcer. Patients with active duodenal ulcer associated with H. pylori infection were randomized to receive omeprazole, 40 mg every morning for 14 days, with either clarithromycin, 500 mg, or placebo three times daily, which was followed by omeprazole, 20 mg every morning for 14 days. Patients underwent endoscopy before enrolling in the study, immediately after finishing treatment, and at 4- to 6-week and 6-month follow-up evaluations or at the recurrence of symptoms. RESULTS: Two hundred and eight patients with active duodenal ulcer associated with confirmed H. pylori infection were randomized to treatment with either clarithromycin and omeprazole (n = 102) or omeprazole and placebo (n = 106). Four to six weeks after treatment was completed, H. pylori was eradicated in 74% (95% confidence interval, 63.0%-82.4%) of patients receiving clarithromycin and omeprazole, compared with 1% (0.0%-6.2%) of patients receiving omeprazole monotherapy (p < .001). Clarithromycin resistance developed in eight patients treated with clarithromycin and omeprazole and in none given omeprazole and placebo. Ulcers, which were healed following treatment in more than 95% of study patients, recurred by the 6-month follow-up visit in 10% (5%-19%) of dual therapy recipients, compared with 50% (39%-61%) of those who took omeprazole alone (p < .001). CONCLUSION: Clarithromycin and omeprazole dual therapy is simple and well tolerated and leads to consistently high eradication rates for patients with duodenal ulcer associated with H. pylori infection. PMID- 9398894 TI - Eradication of Helicobacter pylori using one-week triple therapies combining omeprazole with two antimicrobials: the MACH I Study. AB - BACKGROUND: Eradication of Helicobacter pylori provides potential cure in the majority of patients with peptic ulcer disease, and eradication rates of more than 90% have been reported, using omeprazole in combination with two antimicrobials. The choice of antimicrobials, dose regimen and duration of treatment have varied between studies, however, and an optimal treatment still has to be established. MATERIALS AND METHODS: We conducted an international, randomized, double-blind, placebo-controlled study involving more than 100 patients in each of six treatment groups in 43 hospital gastrointestinal units in Canada, Germany, Ireland, Sweden, and the United Kingdom. Patients (n = 787) with proved duodenal ulcer disease were randomized to treatment twice daily for 1 week with omeprazole, 20 mg (O), plus either placebo (P) or combinations of two of the following antimicrobials: amoxicillin, 1 gm (A), clarithromycin, 250 or 500 mg (C250, C500), or metronidazole, 400 mg (M). Eradication of H. pylori was evaluated by 13C-UBT, performed before and 4 weeks after treatment cessation. RESULTS: The eradication rates for the all-patients-treated analysis were 96%, OAC500; 95%, OMC250; 90%, OMC500; 84%, OAC250; 79%, OAM; and 1%, OP. OAC500 and OMC250 achieved eradication rates with lower 95% confidence interval limits exceeding 90%. All regimens were well-tolerated, 96% of patients complied with their dose regimen, and 2.3% of the patients discontinued treatment owing to adverse events. CONCLUSIONS: Omeprazole triple therapies given twice daily for 1 week produce high eradication rates, are well-tolerated, and are associated with high patient compliance. The two most effective therapies were those combining omeprazole, 20 mg, with either amoxicillin, 1 gm, plus clarithromycin, 500 mg, or metronidazole, 400 mg, plus clarithromycin, 250 mg, all given twice daily. PMID- 9398895 TI - Effectiveness of quadruple therapy using lansoprazole, instead of omeprazole, in curing Helicobacter pylori infection. AB - BACKGROUND: Omeprazole enhances the efficacy of bismuth-based triple therapy. It is unknown whether the same is true for other proton pump inhibitors. Lansoprazole has superior anti-Helicobacter activity in vitro and possibly also in vivo; therefore we investigated quadruple therapy with lansoprazole. MATERIALS AND METHODS: In two studies performed in separate hospitals, a total of 67 Helicobacter pylori-positive patients were treated with 7-day quadruple therapy (lansoprazole, colloidal bismuth subcitrate, tetracycline, and metronidazole) after 3 days of lansoprazole pretreatment. Testing for cure was done by endoscopy in study 1 and by breath test in study 2. RESULTS: Cure rates per protocol were 31 of 31 (100%) in study 1 and 30 of 32 (94%) in study 2. Intention-to-treat cure rates were 31 of 35 (89%) in study 1 and 30 of 32 (94%) in study 2. Cured overall were 32 of 34 with a metronidazole sensitive strain and 3 of 3 with a metronidazole-resistant strain. Data on side effects were collected from 51 patients. Twelve (21%) had no side effects, 27 (53%) had mild side effects, 10 (20%) had moderate side effects, but only 2 (4%) had severe side effects. Side effects, never were the reason that a patient stopped taking the medication. CONCLUSIONS: The results with lansoprazole-quadruple therapy are comparable to the historic control group treated with omeprazole-quadruple therapy. The cure rare is very high, and although mild to moderate side effects occurred in many patients, everybody finished the treatment regime. PMID- 9398896 TI - Helicobacter pylori-positive duodenal ulcer: a long-term double-blind randomized study in patients healed with H2-receptor antagonists. AB - BACKGROUND: The NIH Consensus Conference in 1994 (1) concluded that all patients with peptic ulcer disease should be tested and treated for Helicobacter pylori and that further evaluation was needed for patients in remission. MATERIALS AND METHODS: We evaluated in a double blind randomization 30 patients whose duodenal ulcers had been healed with H2-receptor antagonists and who remained in remission on maintenance therapy. After ulcer healing and the presence of H. pylori had been confirmed, these patients were randomized to receive eradication therapy or placebo and were followed for a mean period of 23 months. RESULTS: Almost all patients receiving placebo had ulcer recurrence, whereas the patients treated with antibiotics demonstrate a low recurrence rate. CONCLUSION: These data suggest, for the first time to our knowledge, the importance of treating with antibiotics duodenal ulcer patients whose disease is in remission. PMID- 9398897 TI - Effect of omeprazole therapy on the survival of Helicobacter pylori, urease activity, and antral gastric histology in patients with duodenal ulcer. AB - BACKGROUND: Helicobacter pylori is associated with chronic active gastritis and peptic ulceration (PU). Omeprazole is a proton pump inhibitor that is effective in healing PU and reducing gastritis. Previously it has been found that omeprazole has some bacteriostatic activity against H. pylori both in vitro and in vivo and in inhibiting urease activity in vitro. Our aim was to evaluate the effect of omeprazole on H. pylori colonization of the gastric mucosa, urease activity in vivo, and the presence of associated gastritis in patients with duodenal ulcer (DU). MATERIALS AND METHODS: We studied 12 patients (7 men and 5 women, ages 22-68 yr) with Du larger than 5 mm in diameter with a positive CLOtest (Delta West Ltd., Australia). Omeprazole, 20 mg bid, was given for 8 weeks to each patient, patients were endoscoped at the end of this period to check for healing of DU, and repeat biopsies were obtained from the gastric antrum for histological analysis, CLOtest, and culture. RESULTS: DU healed completely in all patients. Likewise in all patients there was significant reduction in the urease activity, from 22.1 +/- 4.17 to 1.58 +/- 0.92 units/ml (p < .001; 95% confidence interval of the difference between means, 32.7-14.1), and reduced H. pylori density, from 1,403.46 +/- 128.23 to 422.5 +/- 172.39 colony forming units (CFU) per milligram of tissue biopsy (p < .001; 95% confidence interval of the difference between means, 1,486.1-590.5). The numbers of H. pylori were reduced on the gastric mucosa after omeprazole therapy and disappeared in six patients, a result that correlated with a negative CLOtest reading after 24 hours. CONCLUSION: Omeprazole, 20 mg bid, is capable of reducing H. pylori numbers and urease activity in vivo. There was no significant reduction in the severity of antral gastritis in DU patients studied. PMID- 9398898 TI - Gastric juice polymerase chain reaction: an alternative to histology in the diagnosis of Helicobacter pylori infection. AB - BACKGROUND: Infection from Helicobacter pylori plays a role in several gastroduodenal diseases. The recent availability of molecular techniques, particularly the polymerase chain reaction (PCR), allows us to detect small amounts of this bacterium. The aims of this study were to compare PCR and histological findings and to ascertain the clinical usefulness of H. pylori PCR identification in different biological samples. MATERIALS AND METHODS: We studied 94 consecutive patients. Saliva, gastric juice, and four antral and four body biopsies were obtained from each patient. H. pylori was evaluated histologically in two antral and two body biopsies (Giemsa or Warthin-Starry stain). After extraction, DNA was submitted for PCR amplification using the two primers HPU1 and HPU2, which amplified a 411-bp product from the urease gene A. RESULTS: Forty nine patients were H. pylori-positive at histological workup. The sensitivity of PCR was 92% for gastric juice, 73% for antral biopsies, 61% for body biopsies, and 13% for saliva. Of the 45 H. pylori-negative patients at histological assessment, 7 (16%) had positive findings on PCR, mainly when gastric juice was examined. CONCLUSIONS: These results indicate that PCR is as sensitive as histological assessment. We suggest that PCR H. pylori detection in gastric juice is a sensitive method for diagnosing this infection. PMID- 9398899 TI - Twenty-minute fasting version of the US 13C-urea breath test for the diagnosis of H. pylori infection. AB - BACKGROUND: In large-scale multi-center clinical trials, the US 13C-urea breath test (UBT) has proven to have a sensitivity and specificity of approximately 95%. Ingestion of a meal to delay gastric emptying has advantages of increasing the level of signal as well as prolonging the duration of significantly increased 13C excretion, at the expense of requiring 40 to 60 minutes to complete the test. Our aim was to explore the utility of the 13C-UBT with a total duration of 30 minutes or less. METHODS: After a baseline breath sample was obtained, 125 mg of 13C-urea was given in 100 ml of water, and additional breath samples were taken after 20 and 30 minutes. The results of the UBT were compared to histological assessment, culture, and the rapid urease test. 13C-UBTs were carried out on normal volunteers who underwent gastroscopy during which six mucosal biopsies were taken. Three biopsies were for histological evaluation (Genta stain), two for culture, and one was for agar gel rapid urease testing. The UBT was conducted 2 to 3 days either before or after the endoscopic procedure. RESULTS: The cutoff value for a positive UBT was enrichment of 2.4 delta/1000 (delta over baseline). Of the 66 tests, 51/1000 were Helicobacter pylori-positive. There were no false positive UBTs and only two false negative UBTs at 20 minutes (sensitivity, 96%; specificity, 100%). At 30 minutes, one other UBT was false negative (gray zone of 2.36/1000) (sensitivity, 94%; specificity, 100%). CONCLUSION: These results suggest that omission of the meal and shortening the duration of the US 13C-UBT to 20 minutes still may maintain excellent specificity and sensitivity of the test. PMID- 9398900 TI - Helicobacter pylori infection in congestive gastropathy. AB - BACKGROUND: This study determines the prevalence and significance of Helicobacter pylori infection in portal hypertensive patients. MATERIALS AND METHODS: Patients numbered 118 and consisted of 90 patients with portal hypertension (66 men; 24 women; mean age, 49.1 +/- 2.1 years) and 28 noncirrhotic patients with nonulcer dyspepsia, (12 men; 16 women; mean age, 47.6 +/- 2.8 years), who made up the control group. In all patients, diagnostic upper endoscopy was performed, and gastric biopsies were taken for histological examination and diagnosis of H. pylori. RESULTS: Of the portal hypertensive patients, 42 (47%) had congestive gastropathy, 11 (26%) of whom were positive for H. pylori, and 48 (53%) did not have gastropathy, 12 (25%) of whom were positive for H. pylori. In the control group, 15 of 28 (54%) were positive for H. pylori. H. Pylori was found less frequently in congestive gastropathy patients than in the control group. We found also that the presence and severity of congestive gastropathy is independent of H. pylori status. CONCLUSIONS: We conclude that the role of H. pylori in the pathogenesis of congestive gastropathy is unlikely, and we suggest that there is no need for its routine eradication in cirrhotic patients. PMID- 9398901 TI - Helicobacter pylori infection, gastritis, and the temperature of choice for hot drinks. AB - BACKGROUND: The role of the temperature of the diet as a potential etiological factor for gastritis or peptic ulcer disease has been postulated since the beginning of the century. Animal studies have demonstrated damage to gastric mucosa caused by hot water at 60 to 80 degrees C. In the pre-Helicobacter pylori era it was reported that the majority of ulcer patients preferred hot drinks. It also was reported that the temperature of choice for drinks increased with severity of histological grade of gastritis. We evaluated the association between the preferred temperature of hot drinks and the presence of H. pylori infection. METHODS: We tested the temperature of choice for hot drinking liquids among 12 H. pylori-negative and 43 H. pylori-positive volunteers. We also compared the effect of H. pylori therapy on hot drink temperature preference and, in 32 individuals, whether there was a relation between temperature and the degree of gastric atrophy. RESULTS: There was no difference in the preferred temperature for hot drinks between those volunteers with and without H. pylori infection (63.4 degrees +/- 6 degrees C compared to 61.3 degrees +/- 7 degrees C, respectively) (mean +/- 1 SD, p = .3). There was no change in preferred temperature after successful therapy of the H. pylori infection compared to unsuccessful H. pylori therapy, nor was there a correlation between the preferred temperature and the presence, absence, or degree of gastric atrophy (r2 < 0.001). CONCLUSION: The temperature of preference for hot drinks was not influenced by H. pylori infection or by the presence of atrophic gastritis. PMID- 9398902 TI - Reservoirs of Helicobacter pylori and modes of transmission. PMID- 9398903 TI - Egg passage of rodshaped and coccoid forms of Helicobacter pylori: preliminary studies. AB - BACKGROUND: We used egg passage of bacteria stored in water to evaluate the culturability of the coccoid form of Helicobacter pylori, as a complement to the results obtained from various animal models. Egg passage was performed, as it is a simple, rapid, and well-characterized old method by which to culture and evaluate culturability of bacteria compared to experiments in animal models. Egg passage has been used in such experiments since 1938 for isolation and growth of, for example, Rickettsiae sp. and Chlamydia sp. MATERIALS AND METHODS: The rod shaped form of H. pylori was produced by plate cultures for 4 and 7 days. The coccoid form of H. pylori was produced by culture on agar plates for 10 days, followed by storage in water. These preparations then were inoculated into the yolk sac of differently aged fertilized eggs. RESULTS: Positive culture was obtained from 14 of 17 eggs (82%) inoculated with rod-shaped H. pylori compared to 0 of 22 eggs (0%) inoculated with the coccoid form. CONCLUSION: Culturability of H. pylori is reduced when it converts into the coccoid form produced by starvation and age followed by storage in water for several weeks at room temperature. Egg passage did not raise the culturability of the coccoid form of H. pylori. Our study demonstrates some clear differences between fresh rods and stored cocci forms of H. pylori in terms of culturability when passed through eggs. PMID- 9398904 TI - Hypothesis: Helicobacter toxins and liver. PMID- 9398906 TI - Helicobacter: a new scientific journal. PMID- 9398905 TI - Helicobacter pylori expresses Lewis X. PMID- 9398907 TI - Reflections on the first description of the presence of Helicobacter species in the stomach of mammals. PMID- 9398908 TI - Treatment of Helicobacter pylori infection: a review of the world literature. AB - BACKGROUND: None of the currently used anti-Helicobacter pylori drug regimens cures the infection 100%, and cure results still vary considerably. The present article reviews the effectiveness of currently used antimicrobial regimens, aimed to cure H. pylori infection. METHODS: Data collection started from the beginning of the anti-H. pylori-therapy era until May 1995. No attempt at formal metanalysis has been made, because many studies have been published only in abstract form. Attempts were made to exclude duplicates of studies by comparison to previously reported ones; the authors of suspected duplicates were contacted. After amalgamation of the number of included patients and the number of successfully treated patients, the mean values of eradication rates and the 95% confidence intervals were calculated. RESULTS: A total of 237 treatment arms were analyzed. Bismuth triple therapy continues to reach high eradication rates worldwide (78-89%). Side effects leading to diminished patient compliance and the marked decline of eradication efficacy in cases of metronidazole resistance are considered to be the major drawbacks of this therapy. Proton pump inhibitor (PPI) dual therapy is better tolerated with fewer side effects than is bismuth triple therapy. The mean eradication rates vary from 55 to 75%, and the extremes lie between 24 and 93%. PPI triple therapies have been shown to be very effective against H. pylori (eradication rates, 80-89%). Quadruple therapy leads to a mean eradication rate of 96%. CONCLUSION: Based on efficacy, PPI triple or bismuth triple therapy are recommended as first-line treatment for H. pylori infection. Quadruple therapy could serve as second-line treatment for eradication of initial failures and in case of metronidazole resistance. PMID- 9398909 TI - Immune evasion by Helicobacter pylori: gastric spiral bacteria lack surface immunoglobulin deposition and reactivity with homologous antibodies. AB - BACKGROUND: Helicobacter pylori infection persists in the presence of potent serum and gastric mucosal antibody responses against bacterial antigens. The aim of this article is to report on a study determine whether there is antibody deposition on H. pylori in vivo in the stomach of infected patients and whether gastric and cultured forms of H. pylori differ in their antibody reactivity. MATERIALS AND METHODS: Serum, gastric biopsies, and antral brushings were obtained from 10 patients having endoscopy. H. pylori was cultured from gastric biopsies. Bacterial samples were stained directly for immunoglobulin deposition and indirectly using rabbit antiurease serum or patient serum. Samples were examined by immunofluorescence microscopy and flow cytometry. RESULTS: Although spiral bacteria could be identified easily by acridine orange staining and antiurease staining of gastric brushings from H. pylori infected patients, gastric bacteria did not have detectable IgG or IgA present, and only one of five samples could be stained for IgG and IgA indirectly using patient serum. In contrast, cultured bacteria could be stained readily with homologous serum for IgG and IgA in the majority of cases. Low pH inhibited immunoglobulin reactivity with cultured H. pylori. CONCLUSIONS: Gastric H. pylori may evade humoral defense owing to poor deposition of immunoglobulin in the gastric environment or failure to express surface antigens that are present on cultured forms of H. pylori. PMID- 9398911 TI - Natural and experimental Helicobacter mustelae reinfection following successful antimicrobial eradication in ferrets. AB - BACKGROUND: Recrudescence or reinfection may occur after eradication of Helicobacter pylori in humans. MATERIALS AND METHODS: We used the ferret Helicobacter mustelae model to investigate the effect of prior infection and eradication on reinfection by experimental and natural routes. Two groups of ferrets with naturally acquired H. mustelae infection were treated with an eradication protocol using amoxicillin, metronidazole, and bismuth subsalicylate. The ferrets were monitored for recrudescence by repeated cultures of endoscopic gastric mucosal biopsies. The ferrets were challenged at 17 months (group I) and 6 months (group II) after eradication with a strain of H. mustelae having a distinctive restriction endonuclease analysis pattern. The eradication protocol was repeated to eliminate the infection produced by experimental challenge. The ferrets were then cohoused intermittently with naturally infected ferrets. RESULTS: The original H. mustelae infection was successfully eliminated by the eradication protocol. No recrudescence was observed in group I for 12 months nor for 3 months in group II after eradication. All ferrets became persistently reinfected with the challenge strain. The infection from the challenge strain was eradicated successfully. No ferrets in group I and all ferrets in group II became infected through cohousing. CONCLUSIONS: These results suggest that though prior infection with H. mustelae may confer some protection against reinfection, such protection is not universal in all circumstances; that susceptibility to reinfection by contact with infected animals varies between individuals; and that age may be a factor in this individual variability. These results are applicable to studies of reinfection after eradication of H. pylori in humans. PMID- 9398910 TI - Susceptibility of Helicobacter pylori to the bactericidal activity of human serum. AB - BACKGROUND: Human serum represents an important barrier to the entry of most mucosal organisms into tissues and to the systemic circulation. If at all present, Helicobacter pylori within gastric tissue is rare, and bacteremia for this organism has been described only once. METHODS: To assess the susceptibility of H. pylori to the bactericidal activity present in normal human serum (NHS), we examined 13 H. pylori isolates. To assess the contributions of the classical and alternative complement pathways to killing, we added either C2-deficient or factor B-deficient serum, respectively, to heat-inactivated NHS. Also we assessed the ability of the strains to bind 125I-C3. RESULTS: After incubation for 60 minutes at 37 degrees C, all 13 H. pylori strains were killed by NHS; heating to 56 degrees C for 30 minutes ablated killing, indicating complement dependence for this phenomenon. In the absence of an antibody source, there was no killing when either an alternative or classical complement pathway source was used. Adding B deficient serum to heat-inactivated normal human serum did not restore killing, but adding C2-deficient serum permitted partial killing. All of the 13 strains bound 125I-C3. Although the kinetics varied from strain to strain, C3 bound was significantly correlated (r = 0.61, p = 0.03) with serum susceptibility. CONCLUSIONS: H. pylori are susceptible to complement, alternative pathway activation appears critical, and C3 binding is a major locus of variability. PMID- 9398912 TI - Characterization and therapy for experimental infection by Helicobacter mustelae in ferrets. AB - BACKGROUND: Numerous clinical trials evaluating the efficacy of various antimicrobial compounds against Helicobacter pylori infection have been performed in humans. A convenient animal model for Helicobacter infection would facilitate the evaluation of novel therapies. These experiments were performed to evaluate the use of ferrets as a model of Helicobacter infection. MATERIALS AND METHODS: Ferrets were infected experimentally with Helicobacter mustelae and subsequently treated with bismuth subsalicylate (BSS) triple therapy (BSS, metronidazole, and amoxicillin), or left untreated. The status of infection and serology was assessed during treatment and for 8 weeks posttreatment. Seven ferrets successfully treated with triple therapy were challenged with H. mustelae and monitored for infection for an additional 5 weeks. RESULTS: Infection of ferrets by H. mustelae was accompanied by gastritis and a specific antibody response. Treatment of H. mustelae-infected ferrets with BSS suppressed bacterial growth in four of nine animals but did not eradicate infection. Triple therapy eradicated infection in all nine ferrets with a reduction in gastric inflammation. No relapse of infection occurred up to 8 weeks posttherapy. Challenge with H. mustelae of ferrets successfully treated with triple therapy resulted in a 100% rate of reinfection. CONCLUSIONS: H. mustelae infection can be eliminated by triple therapy, but this does not result in protective immunity against reinfection by H. mustelae. This model, using a strain of Helicobacter indigenous to the host, may be useful for assessing therapeutic efficacy of novel therapies for the treatment of human infection by H. pylori. PMID- 9398913 TI - Atrophic changes of gastric mucosa are caused by Helicobacter pylori infection rather than aging: studies in asymptomatic Japanese adults. AB - BACKGROUND: The current study was designed to evaluate the effect of aging and Helicobacter pylori infection on the gastric mucosa in asymptomatic Japanese adults. MATERIALS AND METHODS: Eighty-five asymptomatic healthy adults were recruited from a health-screening center in Sapporo. All subjects underwent endoscopy and gastric biopsy, and serum was obtained for IgG antibodies to H. pylori, serum gastrin, and pepsinogen levels. RESULTS: The prevalence of atrophic change of the gastric mucosa assessed by pathological findings increased with age (49% in the 30- to 39-year-old group compared to 89% in those 60 years and older, p < .001). The frequency of intestinal metaplasia also increased with age (38% in the 30- to 39-year-old group compared to 82% in those 60 years and older, p < .001). In contrast, the frequency of atrophic gastritis and intestinal metaplasia was extremely low in the H. pylori seronegative group regardless of age. Mean serum gastrin level in H. pylori-positive adults was significantly greater than in those who were H. pylori-negative (114.3 +/- 11.2 compared to 65.8 +/- 6.5 pg/ml, p < .03). The serum pepsinogen I-II ratio was significantly lower in those with H. pylori infection than in those without (3.1 compared to 6.6, p < .0001). CONCLUSIONS: These results suggest that the chronological changes in the gastric mucosa in Japanese individuals are either entirely related to H. pylori infection or the process is greatly accelerated by H. pylori infection. PMID- 9398914 TI - An increase in Helicobacter pylori strains resistant to metronidazole: a five year study. AB - BACKGROUND: Metronidazole is one of the most commonly used antimicrobial agents for the treatment of Helicobacter pylori infection. Resistance to metronidazole has been reported worldwide but with a wide range of prevalence. We started using the classical triple therapy (bismuth, tetracycline, and metronidazole) for H. pylori infection in 1991 but recently have experienced a decline in its efficacy in curing the infection. Thus our aim was to investigate in a single center the prevalence of metronidazole-resistant H. pylori over a period of 5 years. MATERIALS AND METHODS: A total of 1,015 different H. pylori strains collected over a period of 5 years were tested for sensitivity against metronidazole, ampicillin, tetracycline, and imipenem. Antibiotic sensitivity was tested by the disk diffusion and agar dilution methods. To elucidate further the possible relationship between these metronidazole-resistant strains, genomic DNA digestion by the Hae III endonuclease and ribotyping were undertaken in a selected group of isolates. RESULTS: In 1991, 29 of 132 (22.0%) tested strains of H. pylori were found to be resistant to metronidazole. Since our initiation at that time of a triple therapy of bismuth, metronidazole, and tetracycline, the prevalence of metronidazole-resistant strains rose rapidly to 73.2% in 1995. All H. pylori isolates were sensitive to ampicillin, tetracycline, and imipenem. A high degree of genomic heterogeneity was found among these isolates. Thus it is unlikely that the resistant strains of H. pylori were originated from a single clone. CONCLUSIONS: This study shows a rapid increase in metronidazole-resistant H. pylori with the use of an anti-Helicobacter regimen that contains metronidazole. We anticipate that the efficacy of metronidazole-containing anti-Helicobacter regimens will decline with the rapid rise in resistant strains of H. pylori. PMID- 9398915 TI - Culture of Helicobacter pylori: effect of preimmersion of biopsy forceps in formalin. AB - BACKGROUND: Treatment of antibiotic-resistant Helicobacter pylori should be based on bacterial sensitivity testing that requires the ability to isolate the bacterium from gastric mucosal biopsies. The aim of this study was to determine whether the yield for detecting H. pylori infection by culture is reduced by immersion of biopsy forceps in formalin prior to obtaining the specimen. MATERIALS AND METHODS: Gastric antral mucosal biopsies (100 specimens) from 50 patients were obtained for culture of H. pylori. An antral biopsy was taken for culture, and with the same forceps a biopsy was taken for histological examination. The biopsy specimen was removed by shaking, whereas the forceps was immersed in 10% buffered formalin for the histological investigation. The forceps was then used without rinsing to obtain a second specimen for culture from an area adjacent to the first site. H. pylori status was determined by histological assessment with the Genta stain and a rapid urease test. RESULTS: Fifty patients with H. pylori infection documented by histological inquiry and positive rapid urease testing entered the study; 29 had duodenal ulcers, 5 had gastric ulcers, 1 had mucosal associated lymphoid tissue (MALT) lymphoma, and 15 were without ulcer disease. The results of culture both before and after immersion in formalin were identical. One patient had both cultures negative; the sensitivity of culture for detection of H. pylori infection was 98% (95% confidence interval = 93%-100%). CONCLUSION: Preimmersion of biopsy forceps in formalin does not adversely affect the ability to culture H. pylori. PMID- 9398916 TI - The influence of Helicobacter pylori status on circulating levels of the coagulation factors fibrinogen, von Willebrand factor, factor VII, and factor VIII. AB - BACKGROUND: Helicobacter pylori (H. pylori) infection has been associated with an increased risk of developing ischemic heart disease (IHD). It has been suggested that a persisting low-grade acute phase response results from the chronic inflammation caused by H. pylori infection, which may give rise to increased circulating levels of certain coagulation factors. MATERIALS AND METHODS: One hundred three (53 male) nonconsecutive, randomly selected white subjects with symptoms of dyspepsia were recruited for study from an outpatient endoscopy clinic at Leeds General Infirmary. The presence of H. pylori was determined by histological and microbiological investigation, a rapid urease test, and a 13carbon urea breath test (13C-UBT). Fibrinogen was measured by the Clauss method, factor VIII:C (FVIII:C) and factor VII:C (FVII:C) were measured by clotting rate assays, and the von Willebrand factor (vWF) was determined by an enzyme-linked immunosorbent assay. RESULTS: No difference was found in levels of coagulation factors according to H. pylori status. Multiple regression models were used to account for the effect of covariates and H. pylori status on levels of FVII:C, FVIII:C, vWF, and fibrinogen, and again H. pylori status was not a significant determinant of levels of any of these coagulation factors. No difference occurred in full blood count, platelet count, white cell count, or plasma viscosity in individuals who were H. pylori-positive compared with those who were negative. CONCLUSIONS: H. pylori infection is not associated with increased circulating levels of fibrinogen, FVII:C, vWF.Ag, or FVIII:C or hemorrheology in this patient group. PMID- 9398918 TI - Sodium cromoglycate ophthalmic solution as a Pharmacy Medicine for the management of mild-to-moderate, non-infectious inflammation of the conjunctiva in adults. PMID- 9398917 TI - Assessment of vision behind cataracts. PMID- 9398919 TI - Gonioscopy: evaluation of the anterior chamber angle. Part I. PMID- 9398920 TI - The clinical features and classification of diabetic retinopathy. PMID- 9398922 TI - Has head and neck surgery a real future? PMID- 9398921 TI - Fibroblasts obtained from human nasal, laryngeal and tracheal mucosa produce the chemokine RANTES. AB - RANTES is a chemokine that was already found in tissues obtained from nasal polyps of patients suffering from chronic polypous sinusitis and in lung biopsies of patients suffering from bronchial asthma. Nasal fibroblasts could be shown to be a cellular origin of RANTES. The aim of this study was to investigate whether human nasal, laryngeal and tracheal mucosa fibroblasts are differentiated in production of RANTES. Fibroblasts obtained from healthy human nasal, laryngeal and tracheal mucosa, were cultured. Secretion of RANTES-protein in supernatants was investigated after stimulation with 50 ng/ml tumor necrosis factor-alpha (TNF alpha), interleukin-1-beta (IL-1-beta), lipopolysaccharide (LPS), phorbolmyrisate acetate (PMA), interferon-gamma (IFN-gamma) and serum-free medium (SFM) for 24 hours. Cultivated nasal, laryngeal and tracheal fibroblasts secreted RANTES protein upon TNF-alpha, IL-1-beta and IFN-gamma stimulation. The amounts of RANTES-protein production ranged from 10 ng/ml (PMA) to 198 ng/ml (TNF-alpha). Secretion of significant amounts of RANTES-protein were detected in the supernatants from either nasal, laryngeal or tracheal fibroblasts. There was no significant difference between the differential fibroblasts. We conclude that nasal, laryngeal and tracheal fibroblasts could be a cellular source of RANTES in nasal and bronchial mucosa or in secrets of patients suffering from diseases where eosinophilic tissue infiltration represents a characteristic histopathological feature. Results suggest that additional local factors are needed to develop asthma bronchiale and chronic polypous sinusitis. PMID- 9398923 TI - Assessment of genetic susceptibility as a risk factor for head and neck squamous cell carcinoma. PMID- 9398924 TI - Minimal invasive ear, nose and throat surgery--advances through modern technologies. AB - Three fundamentals have to be fulfilled to optimize minimally invasive surgery: three-dimensional imaging, free maneuverability of the instruments, sensorial feedback. Projection of two pictures from a stereoendoscope and subsequent separation with a LCD shutter allows three-dimensional videoendoscopy to be performed. A high frequency shutter technique (100/120 Hz) presents pictures from the two video cameras to the right and left eye, respectively, so that the surgeon has spatial vision of the operative field. Steerable instruments have four components: a control unit, rigid shaft, steerable multijoint, distal effector. The steerable multijoints give two additional degrees of freedom compared to conventional rigid instruments in endoscopic surgery. For intuitive movements, however, an electronic control system is necessary that is comparable to the "master slave" priniciple in remote technology. A remote manipulator system with six degrees of freedom is now available. Additionally, a multifunctional distal tip permits different surgical steps to be performed without changing the instrument. For better control of the instrument and the operative procedure tactile feedback can be achieved with appropriate microsensor system. Recent projects suggest that an artificial sensor system can be established within the foreseeable future. PMID- 9398925 TI - The link between substance abuse and posttraumatic stress disorder in women. A research review. AB - Research has documented a high incidence of comorbid post-traumatic stress disorder (PTSD) and substance abuse. Women substance abusers, in particular, show high rates of this dual diagnosis (30% to 59%), most commonly deriving from a history of repetitive childhood physical and/or sexual assault. Rates for men are two to three times lower and typically stem from combat or crime trauma. Patients with both disorders are characterized by high severity on a multitude of psychological and treatment variables and use of the most severe drugs (cocaine and opioids). Treatment research on women is limited but suggests the possibility of retaining patients and achieving positive outcomes. PMID- 9398926 TI - Course and severity of substance abuse among patients with comorbid major depression. AB - The authors assessed the course and severity of substance-related disorder (SRD) among patients with comorbid major depressive disorder (MDD) by means of both retrospective and concurrent data. A total of 642 patients were assessed. Data on course included lifetime use, age at first use, years of use, use in the last year; periods of abstinence, and current diagnosis. Data on severity included two measures of SRD-associated problems, substance abuse vs. dependence, self-help activities, and number of substances being abused. SRD-MDD patients tended to manifest lower levels of cannabis, opiate, and cocaine use, and more SRD-only patients were abusing three or more substances. Men with SRD-MDD demonstrated longer mean durations of abstinence compared with men with SRD-only, whereas SRD MDD women demonstrated shorter mean durations of abstinence, compared with women with SRD-only. MDD-SRD patients showed slightly less substance abuse, but SRD severity was comparable with SRD-only patients. PMID- 9398927 TI - Retention in substance abuse treatment. Role of psychiatric symptom severity. AB - The authors investigated the association between psychiatric symptom severity and subsequent treatment retention among substance abusers. The Symptom Check List-90 R was administered, after admission to an addiction treatment facility, to 308 male and 106 female clients with moderate-to-severe substance abuse problems. Mean scores on nine symptom and three summary scales were computed. Controlling for other sociodemographic and treatment variables, somatization was significantly associated with dropout from specialized outpatient and inpatient treatment programs. This study, however, suggests that psychopathologic symptom severity at admission has only limited utility in predicting subsequent treatment retention among substance abusers with overall moderate levels of psychological distress. PMID- 9398928 TI - Suicide and alcoholism. Distinguishing alcoholic patients with and without comorbid drug abuse. AB - Psychological autopsy data were used to test the hypothesis that alcoholic patients with comorbid drug use disorders who committed suicide (A + D; n = 26) are distinguishable from alcoholic suicide victims without a comorbid drug use disorder (A; n = 35). Dependent variables included demographics, suicidal behavior; psychiatric symptoms, and medical illness burden. The A group were older, white, and tended to be living alone. Analyses that controlled for age and sex indicated that As were more likely to have had a comorbid major depression and less likely to tell someone they were contemplating suicide. Scores on a measure of illness burden increased with age among the A group but not the A + D group, though the latter were more likely to be under a physician's care with increasing age. These differences should be considered when designing preventive measures. PMID- 9398929 TI - Differences between men and women in dual-diagnosis treatment. AB - The authors reviewed the charts of all women and a randomly selected sample of men over a 6-month period on two addiction treatment units at Bellevue Hospital Center in New York. The men were more likely to be admitted with schizophrenia and to have used substances of abuse other than alcohol, and the women were more likely to be admitted with affective disorders. Also, the women on the dual diagnosis ward were more likely to be domiciled (i.e., not homeless), and the women on both units were significantly more likely to report having been crime victims. These findings suggest that dually diagnosed women need a substantially different treatment paradigm from men. PMID- 9398930 TI - Associations between ADHD and psychoactive substance use disorders. Findings from a longitudinal study of high-risk siblings of ADHD children. AB - This article investigates the relationship between attention deficit/hyperactivity disorder (ADHD) and psychoactive substance use disorders (PSUD) in siblings of ADHD and normal-control probands and addresses issues of psychiatric comorbidity and gender. Using DSM-III-R structured diagnostic interviews and blind raters, the authors conducted a 4-year follow-up of siblings. ADHD and male gender predicted higher rates and an earlier onset of PSUD after adjusting for high-risk status, other psychiatric disorders, and age. Risk was particularly high if the siblings had ADHD plus conduct disorder. This study's findings confirms the authors' prior report high-lighting the importance of drug and alcohol prevention and cessation programs aimed at ADHD youth and their siblings, particularly those with comorbid conduct disorder. PMID- 9398931 TI - Managing disability benefits as part of treatment for persons with severe mental illness and comorbid drug/alcohol disorders. A comparative study of payee and non payee participants. AB - The objective of this pilot study is to describe the use of a Social Security representative payee program as a clinical intervention integrated into long term, dual-disorder treatment of severely mentally ill outpatients with comorbid drug/alcohol disorders. Compared with non-payees, patients selected to be payee participants were more likely to be male, have a diagnosis of schizophrenia, have a history of high inpatient utilization, and have higher current ratings of psychiatric symptoms, substance use, and functional disability. Despite these higher severity ratings, which usually predict poor outpatient compliance and higher rate of adverse outcomes, the payee participants attended about twice the number of outpatient service sessions as non-payees and were no more likely to be currently homeless, hospitalized, or incarcerated. The payee intervention is described, and ethical and research issues are discussed. PMID- 9398932 TI - Methadone vs. L-alpha-acetylmethadol (LAAM) in the treatment of opiate addiction. A meta-analysis of the randomized, controlled trials. AB - The authors conducted a meta-analysis of the reported randomized, controlled trials comparing methadone to L-alpha-acetylmethadol (LAAM) to assess the efficacy of LAAM relative to methadone in the treatment of opiate addiction. All studies were conducted in standard outpatient opiate addiction treatment clinics. Most patients were men from lower socioeconomic strata. A statistically significant risk difference favoring methadone was detected for retention-in treatment and discontinuation of treatment because of side effects. The risk difference for illicit drug use favored LAAM, but was not significant. A small treatment difference in favor of methadone was noted. LAAM does appear to be a relatively effective alternative in the treatment of opiate addiction, more so in certain select situations. PMID- 9398934 TI - Addressing epistemologic and practical issues in multimethod research: a procedure for conceptual triangulation. AB - Although a combination of quantitative and qualitative methods are being used increasingly in nursing research, little practical advice exists about the conduct of such studies. Conceptual triangulation offers one approach to multimethod research, addressing epistemologic and practical issues that have long plagued investigators. Conducting quantitative and qualitative research as parallel studies and using method-specific criteria for rigor provide an alternative to blending methodologic assumptions. Systematic examination of the support for findings guides judgments about model development, and the provision for multiple conceptual models resolves issues about the interpretation of findings. PMID- 9398935 TI - Constructivism: a naturalistic methodology for nursing inquiry. AB - This article will explore the philosophical underpinnings of the constructivist research paradigm. Despite its increasing popularity in evaluative health research studies there is limited recognition of constructivism in popular research texts. Lincoln and Guba's original approach to constructivist methodology is outlined and a detailed framework for nursing research is offered. Fundamental issues and concerns surrounding this methodology are debated and differences between method and methodology are highlighted. PMID- 9398936 TI - Expanding the praxis debate: contributions to clinical inquiry. AB - Nursing science continues to debate the adequacy of various philosophic paradigms for their ability to forward the discipline. Nursing must embrace multiple paradigms, methodologies, and their philosophic assumptions to adequately address the complex and multifaceted human phenomena that is the focus of clinical inquiry in nursing. This article examines the differences in interpretive and critical approaches to clinical inquiry relative to praxis, expanding how praxis can be used to inform nursing practice. Differences in the nature of knowledge, goals of inquiry, and claims to praxis between the interpretive and critical traditions are discussed. Praxis, realized through clinical inquiry in both the interpretive and the critical paradigms, may contribute important pieces of the puzzle to improve the human condition. Expanding the praxis debate challenges nurses to consider the emancipatory possibilities of clinical inquiry within both interpretive and clinical paradigms. PMID- 9398937 TI - Advocacy oral history: a research methodology for social activism in nursing. AB - The reinstatement of social activism as a central feature of nursing practice has been advocated by nursing scholars and is consistent with contemporary conceptualizations of primary health care and health promotion that are rooted in critical social theory's concept of empowerment. Advocacy oral history from a feminist postmodern perspective offers a method of research that has the potential and purpose to empower participants to transform their political and social realities and may, therefore, be considered social activism. A recent study of public health nurses who had experienced significant distress through the reduction and redirection of their practice is provided as an exemplar of advocacy oral history. Philosophies underpinning the research method and characteristics of feminist postmodern research are reviewed and implications for the use of this methodology for social activism in nursing are drawn. PMID- 9398938 TI - The facilitator in participatory action research: les raisons d'etre. AB - Participatory action research (PAR) is a form of qualitative inquiry that posits a collaborative, self-reflective process among its participants to develop knowledge. The researcher, along with the participants in this critical research process, identify and illustrate their individual concerns, beliefs, and values. The role of the researcher, as facilitator, is an extremely important social and political position within the participant group, within nursing, and within society. However, how the nurse as facilitator initially "guides" the discussions within the participatory group is a contentious issue for researchers involved in PAR. PMID- 9398939 TI - Summary oral reflective analysis: a method for interview data analysis in feminist qualitative research. AB - This article explores an innovative approach to qualitative data analysis called Summary Oral Reflective Analysis (SORA). The method preserves the richness and contextuality of in-depth interview data within a broader feminist philosophical perspective. This multidisciplinary approach was developed in two individual research programs within a cooperative, collaborative arrangement. It represents a creative response to perceived deficiencies in the pragmatics of qualitative data analysis where the maintenance of data contextuality is critical. PMID- 9398940 TI - Secondary analysis of qualitative data. AB - Secondary analysis of qualitative data is a valid mode of clinical inquiry. However, there is limited information available on its use in nursing. This article describes the use of secondary analysis for a study of family caregivers of relatives with dementia. The advantages, limitations, and application of secondary analysis are outlined; data management, analysis, and rigor are also discussed. The article concludes that this method is cost-effective, decreases respondent burden, and is a useful research method for students. However, the secondary analyst must be aware of the limitations of using secondary analysis of qualitative data. PMID- 9398941 TI - The responsive use of self in community health nursing practice. AB - This interpretive study examined the expertise that is often unrecognized in the everyday practice of community health nurses. Twenty-five nurses participated in the study and were asked to describe meaningful clinical situations during group and individual interviews. Field notes of observations of clinical situations and transcribed interviews were analyzed as a text. A major finding of the study involved the nurse's responsive use of self. Responsiveness to the other enabled the nurse to gain a situated understanding of clients' lives and to cultivate clients' strengths and connections to a responsive community. PMID- 9398943 TI - The mechanism for the effect of selenium supplementation on immunity. AB - Lipid peroxide (LPO) in lymphocytes from mice was evaluated by measuring substances reactive to thiobarbituric acid (TBA). The product resulting from the reaction of TBA with lymphocytes was extracted with n-butyl and fluorescence intensity was determined. The degree of lipid peroxidation, expressed as fluorescence intensity f547, was assessed for stimulation of lymphocytes with concanavalin A (Con A), and was related to lymphocyte proliferation in response to Con A if Se was administered. The lymphocyte proliferation was determined by [3H]thymidine incorporation, expressed as cpm. The effect of superoxide dismutase (SOD), added to cell culture on lymphocyte proliferation was also evaluated. It was found that LPO in lymphocytes before Con A stimulation was significantly less than that after stimulation (p < 0.001), and that SOD promoted lymphocyte proliferation dose dependently. The addition of Na2Seo3 to lymphocyte culture or supplementation in drinking water to mice decreased the produced LPO in lymphocyte in response to Con A. In the presence of Se, there is an inverse correlation between the levels of LPO in lymphocyte and the stimulated proliferation (r = -0.8902, r = -0.9439). In conclusion, active oxygen species scavenging was proposed as one of the mechanisms for Se to promote immunity. PMID- 9398942 TI - Activities of liver microsomal fatty acid desaturases in zinc-deficient rats force-fed diets with a coconut oil/safflower oil mixture of linseed oil. AB - The present study was conducted to investigate the effect of zinc deficiency on fatty acid desaturation in rats fed two different types of dietary fat, a mixture of coconut oil and safflower oil (7:1, w/w, "coconut oil diet") or linseed oil ("linseed oil diet"). In order to ensure an adequate food intake, all rats were force-fed by gastric tube. Zinc deficiency caused statistical significant reduction of delta 9-desaturase activity in liver microsomes of rats fed coconut oil diet and tendencial reduction (p < 0.15) in rats fed linseed oil diet compared with control rats fed diets with the same type of fat. In agreement with this effect, zinc deficiency in the rats fed both types of dietary fat increased the ratio between total saturated and total monounsaturated fatty in liver phospholipids and liver microsomes. Zinc deficient rats on the coconut oil diet had unchanged delta 6-desaturase activity with linoleic acid as substrate and lowered activity with alpha-linolenic acid as substrate. In contrast, zinc deficient rats on the linseed oil diet had increased delta 6-desaturase activity with linoleic acid as substrate and unchanged activity with alpha-linolenic acid. Because linoleic acid is the main substrate for delta 6-desaturase in the rats fed coconut oil diet, and alpha-linolenic acid is the main substrate in the rats fed linseed oil diet, it is concluded that in vivo delta 6-desaturation was not changed by zinc deficiency in the rats fed both types of dietary fat. Activity of delta 5-desaturase was also not changed by zinc deficiency in the rats fed both dietary fats. Levels of fatty acids in liver phospholipids and microsomes derived by delta 4-, delta 5-, and delta 6-desaturation were not consistently changed by zinc deficiency in the rats fed both types of dietary fat. Thus, the enzyme studies and also fatty acid composition data of liver phospholipids and microsomes indicate that zinc deficiency does not considerably disturb desaturation of linoleic and alpha-linolenic acid. Therefore, it is suggested that similarities between deficiencies of zinc and essential fatty acids described in literature are not due to disturbed desaturation of linoleic acid in zinc deficiency. The present study also indicates that zinc deficiency enhances incorporation of eicosapentaenoic acid into phosphatidylcholine of rats fed diets with large amounts of n-3 polyunsaturated fatty acids. PMID- 9398944 TI - Effect of selenium in recovery of immunity damaged by H2O2 and 60Co radiation. AB - Con A stimulated lymphocytes proliferation was measured as [3H]thymidine incorporation and IgG was quantified by single radial immunodiffusion to study recovering or protecting effect of selenium (Se) on immunity attacked by exogenous active oxygen species, H2O2 and 60Co-radiation, respectively. Lipid peroxidation was also determined to observe the relation between antioxidation ability and protecting ability of Se. It was found that H2O2 injured lymphocytes immunocompetence deeply and 60Co-radiation decreased immune response capacity greatly, but that administration of Se counteracts this damage. The antioxidative ability of Se was correlated with its protecting ability. PMID- 9398945 TI - Chemical speciation of arsenic in urine of patients with blackfoot disease. AB - Blackfoot disease is a peripheral vascular disease resulting in gangrene of the lower extremities. Although extensive epidemiological study has implicated high arsenic content in artesian well water of the endemic area bears some important connection with the disease, the etiology of the disease is still not clarified. In this study, attention is paid to chemical speciation of arsenic in order to find out whether the concentrations of arsenic species in urine of Blackfoot disease patients are different from those of controls. Experimental results indicate that the total arsenic, inorganic arsenic, monomethylarsonic acid, and other forms of arsenic in the urine of patients are significantly higher than those of the controls. The possible connection of those arsenic species with the etiology of the disease is discussed. PMID- 9398946 TI - Trace element concentration and arsenic speciation in the well water of a Taiwan area with endemic blackfoot disease. AB - Blackfoot disease is a peripheral vascular disease resulting in gangrene of the lower extremities. Although extensive epidemiological study has implicated high arsenic content in artesian well water in the endemic area, there is more to learn about the etiology of the disease. In this study, effort is paid on multielement determination and arsenic speciation in order to find out whether the trace element concentration pattern in well water in the Blackfoot disease endemic area is different from those of two control areas. Experimental results indicate that the concentrations of Fe, P, Na, and Ba in well water in the Blackfoot disease endemic area are found to be significantly higher than those of the controls, but they are still below the drinking water standard. The total arsenic in well water in the endemic area (671 +/- 149 ppb) is much higher than that of one normal control area of Hsin-Chu (< 0.7 ppb), but is a similar level as that of other control areas of I-Lan (653 +/- 71 ppb) where no Blackfoot disease has ever been found. It was also found that the insoluble arsenic in the endemic area (21.9 ppb) is much higher than that in two control areas (< or = 1.8 ppb), and the concentration ratio between As(III) and As(V) species in the endemic area (2.6) is much lower than that in one of the control areas, where the total arsenic is also high (14.7). The possible connection of Blackfoot disease with trace elements, arsenic species, and possibly other as yet undefined environmental factors in the artesian well water, is discussed. PMID- 9398947 TI - Time course effects of vanadium supplement on cytosolic reduced glutathione level and glutathione S-transferase activity. AB - The influence of vanadium, an important dietary micronutrient, was evaluated on the cytosolic reduced glutathione (GSH) content and glutathione S-transferase (GST) activity in several rat target tissues. Supplementation of drinking water with vanadium at the level of 0.2 or 0.5 ppm for 4, 8, or 12 wk was found to increase the GSH level with a concomitant elevation in GST activity in the liver followed by small intestine mucosa, large intestine mucosa, and kidney. The results were almost dose-dependent and mostly pronounced with 0.5 ppm vanadium after 12 wk of its continuous supplementation. Neither the GSH level nor GST activity was significantly altered in forestomach and lung following vanadium supplementation throughout the study. The levels of vanadium that were found to increase the content of GSH and activity of GST in the liver, intestine, and kidney did not exert any toxic manifestation as evidenced from water and food consumption as well as the growth responses of the experimental animals. Moreover, these doses of vanadium did not impair either hepatic or renal functions as they did not alter the serum activities of glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), sorbitol dehydrogenase (SDH), as well as serum urea and creatinine level. All these results clearly indicate that vanadium under the doses employed in our study has a significant inducing role on GSH content with a concurrent elevation in GST activity in the liver and specific extrahepatic tissues without any apparent sign of cytotoxicity. This attribute of vanadium may have a greater importance in terms of biotransformation and detoxification of xenobiotics, including carcinogens. In addition, since the ability to afford an increment in the endogenous GSH-GST pool by anticarcinogenic natural substances has been found to correlate with their activity to inhibit neoplastic transformation, the trace element vanadium may be considered as a novel anticancer agent. PMID- 9398949 TI - Neuromuscular relaxants--1997. PMID- 9398948 TI - Effects of nickel oxide on the production of tumor necrosis factor by alveolar macrophages of rats. AB - To evaluate the effect of green nickel oxide (NiO) on the production of tumor necrosis factor (TNF) by alveolar macrophages, alveolar macrophages were exposed to NiO in vitro and in vivo. For the in vitro study, rats alveolar macrophages were incubated with NiO on a microplate for 24 h. TNF activity in the culture supernatant was determined by the L929 bioassay. Rats alveolar macrophages cultured with 100 and 200 micrograms/mL of NiO in vitro induced the production of TNF, however, it was not statistically significant compared with the control that was free from NiO exposure. For exposure in vivo, rats were divided into two groups. Five were exposed to a daily concentration of 11.7 +/- 2.0 mg/m3 of NiO for an 8-hr/d, 5 d/wk, for 4 wk, and five rats (control) were kept in a cage and not exposed to NiO. Bronchoalveolar lavage was performed and the recovered alveolar macrophages were incubated on a microplate for 24 h. TNF production by exposed alveolar macrophages was significantly higher than that of controls. PMID- 9398950 TI - Comparison of vecuronium with ORG 9487 and their interaction. AB - PURPOSE: To compare the pharmacodynamic behaviour of vecuronium with that of ORG 9487, we measured the time-course of action of equipotent doses of ORG 9487 and vecuronium and investigated their mutual interaction when given in succession. METHODS: Sixty ASA I-II patients were anaesthetized with thiopentone, fentanyl, halothane and nitrous oxide and assigned randomly to four groups. Each patient received an initial dose (ID) of either vecuronium (V) or ORG 9487 (O) followed by maintenance doses (MDn) of either V or O (ID/MD: O/O, V/O, O/V, and V/V). The time course of action was measured mechanomyographically, determining the duration until 25% recovery of the single twitch (DUR25). RESULTS: The onset time of an ID of ORG 9487 was shorter than that of an ID of vecuronium (96 vs 203 sec, P < 0.001). The DUR25 of the ID of ORG 9487 was less than half that of vecuronium (10.7 +/- 2.8 vs 28.8 +/- 6.1 min, P < 0.001). The DUR25 of MD1 and MD2 of ORG 9487 were shorter than those of vecuronium (O/O: 7.3 +/- 2.8 and 8.5 +/- 2.4 min; V/O: 12.7 +/- 3.3 and 11.5 +/- 3.5 min, vs O/V: 16.4 +/- 4.5 and 20.6 +/- 4.7 min; V/V: 18.8 +/- 3.0 and 20.1 +/- 3.8 min, respectively, P < 0.05). AN ID of vecuronium prolonged the DUR25 of MD1 and MD2 of ORG 9487 (P < 0.05). CONCLUSION: ORG 9487 is a muscle relaxant with a shorter duration of action than vecuronium. Maintenance doses of ORG 9487 are also shorter acting than roughly equipotent maintenance doses of vecuronium, irrespective of which relaxant is given initially. PMID- 9398951 TI - Comparison of rocuronium and d-tubocurarine for prevention of succinylcholine induced fasciculations and myalgia. AB - PURPOSE: We compared d-tubocurarine and rocuronium for the prevention of succinylcholine-induced fasciculations and postoperative myalgia (POM) and evaluated the influence of both drugs on the speed of onset and recovery of succinylcholine. METHODS: Seventy-five women undergoing surgery of short duration were studied. They were randomized to one of three groups: group SAL received normal saline followed three minutes later by 1.0 mg.kg-1 succinylcholine; group ROC received 0.05 mg.kg-1 rocuronium + 1.5 mg.kg-1 succinylcholine; group DTC received 0.05 mg.kg-1 d-tubocurarine + 1.5 mg.kg-1 succinylcholine. Single-twitch stimulation was applied to the ulnar nerve every 10 sec and the EMG response of the adductor pollicis was recorded. Fasciculations were assessed by a blinded observer on a scale of 0-3. Patients were asked 24 and 48 hr later to rate POM using a scale of 0-10. RESULTS: The interval needed for twitch height to decrease to 10% of initial value after succinylcholine was longer in group ROC (58 +/- 20 sec) (mean +/- SD) compared with group SAL (44 +/- 13 sec) (P < 0.05). Recovery to 20% occurred faster in group ROC (324 +/- 83 sec) than in groups SAL (456 +/- 103 sec) and DTC (450 +/-132 sec) (P < 0.05). Fasciculations were more intense in groups SAL than in groups ROC and DTC (P < 0.001). Patients rated POM as less intense 24hr postoperatively only in group ROC (1.2 +/- 2.4) compared with group SAL (3.3 +/- 3.5) (P < 0.05). CONCLUSION: Rocuronium prevents succinylcholine induced fasciculations and POM. Rocuronium also delays the onset of succinylcholine and shortens its duration compared with d-tubocurarine. PMID- 9398952 TI - Physostigmine increases the dose of propofol required to induce anaesthesia. AB - PURPOSE: This prospective, randomized, double-blind study was performed to determine the effect of administration of physostigmine on the dose of propofol required to produce loss of consciousness. METHODS: Forty female unpremedicated patients were assigned in a random blind design to receive either 2 mg physostigmine or equal volume of normal saline i.v. five minutes before induction of anaesthesia with propofol. All patients received general anaesthesia for breast surgery. Propofol was infused at a constant rate of 200 ml.hr-1 while patients were breathing oxygen 100% via a face mask. In each patient the dose of propofol required to produce loss of the ability to grasp a 20 ml syringe was recorded as the end-point of loss of consciousness. At this point the protocol was terminated and, after intubation of the trachea, anaesthesia was maintained with a nitrous oxide-isoflurane or sevoflurane mixture in oxygen, increments of an opioid and a muscle relaxant. Doses of anaesthetic drugs and duration of anaesthesia varied and depended on the type of breast surgery, determined by frozen section. RESULTS: The mean +/- SD dose of propofol required to produce loss of consciousness was 2.4 +/- 0.6 mg.kg-1 and 2.0 +/- 0.4 mg.kg-1 in the physostigmine and in the normal saline groups respectively (P = 0.014). CONCLUSION: Physostigmine pretreatment increases the dose of propofol required to produce loss of consciousness. PMID- 9398953 TI - Pharmacoeconomics of intravenous regional anaesthesia vs general anaesthesia for outpatient hand surgery. AB - PURPOSE: To compare the cost and effectiveness of intravenous regional anaesthesia (IVRA) with general anaesthesia (GA) for outpatient hand surgery. METHOD: A retrospective record analysis of 121 patients who received IVRA were compared with 64 patients who received GA in our Daycare centre. The costs of anaesthesia and recovery were calculated from an institutional perspective using 1995 Canadian Dollar values. Effectiveness was measured in terms of time for anaesthesia, recovery and discharge, % with unsatisfactory anaesthesia and complications. RESULTS: Both groups were well matched in terms of weight, sex and ASA class. Patients in the IVRA group were older (45 +/- 16 vs 38 +/- 13 yr) and had a lower frequency of two types of operation. The median total cost for the IVRA group of $24.60 (15.76-55.29) was less than that for the GA group of +f448.66 (35.59-73.11), (P < 0.00001). Anaesthesia was unsatisfactory in 11% of the IVRA group, but in none having GA,(P < 0.01). Recovery was faster in the IVRA group with a median time to discharge of 70 (35-180) min compared with 118 (45 320) min in the GA group, (P < 0.00001). Vomiting requiring treatment occurred in 5% of the GA group, but in none having IVRA, (P < 0.05). Dizziness which delayed discharged also occurred in 5% of the GA group, but in none having IVRA, (P < 0.05). CONCLUSION: The cost of anaesthesia and recovery using IVRA for outpatient hand surgery was half that of GA. intravenous regional anaesthesia was less effective than GA in achieving satisfactory anaesthesia, equally effective in time to administer anaesthesia, and more effective in speeding recovery and minimising postoperative complications. PMID- 9398954 TI - Platelet aggregation is impaired during anaesthesia with sevoflurane but not with isoflurane. AB - PURPOSE: Halothane suppresses platelet aggregation in vitro and ex vivo, and prolongs bleeding time. In a previous in vitro study we demonstrated that sevoflurane had a more suppressive effect on platelet aggregation than did halothane. The present study investigated whether the clinical use of sevoflurane affected platelet aggregation ex vivo. METHODS: Thirty-eight patients undergoing minor elective surgery were divided randomly into sevoflurane and isoflurane groups. Anaesthesia was induced with thiopentone i.v., and was maintained with sevoflurane or isoflurane with nitrous oxide. Blood was collected to measure platelet aggregation induced by adenosine diphosphate (ADP) and epinephrine. The first (control) blood collection was performed in the operating room before induction of anaesthesia, and the second 5-10 min after tracheal intubation but before the start of surgery, when the end-expiratory sevoflurane or isoflurane concentrations had stabilised at 1-1.5 times the minimum alveolar concentration (MAC) and mean arterial pressures were between 80-120% of preanaesthetic values. RESULTS: In all samples obtained during sevoflurane anaesthesia (n = 15), ADP and epinephrine could not induce secondary aggregation, although they did induce primary aggregation. In contrast, in the isoflurane group, both primary and secondary aggregation were observed in 14 out of 15 patients, and secondary aggregation was abolished in only one of the samples obtained during anaesthesia. CONCLUSIONS: Sevoflurane, but not isoflurane, alters platelet aggregation in the clinical situation, possibly by suppression of thromboxane A2 formation. PMID- 9398955 TI - Arterial oxygenation during one lung ventilation. AB - PURPOSE: To compare the effects of isoflurane and sevoflurane on arterial oxygenation and middle cerebral artery blood flow velocity during one lung ventilation. METHODS: This was a randomized, crossover study in 20 patients undergoing thoracotomy for oesophageal cancer and scheduled for long term one lung ventilation (OLV). They were randomized to one of two groups: group A, firstly isoflurane was administered followed by sevoflurane, and then isoflurane was resumed; group B, the order of the administration was reversed. Arterial blood gas samples were drawn at the start of OLV, 30 and 60 min after the initiation of OLV and the end of OLV (the change of volatile anesthetics was done 30 and 60 min after the start of OLV). Middle cerebral artery (MCA) was monitored continuously with the probe positioned over the temporal bone window. This probe transmitted 2 MHZ wave Doppler signals. Time-averaged MCA blood flow velocity was calculated from the signals. RESULTS: The PaO2 values decreased 30 min after the start of OLV (364.4 +/- 33.4 mmHg vs 179.0 +/- 19.5, and 338.7 +/- 24.8 mmHg vs 139.7-19.9 in groups A and B respectively), but there was no difference between the groups. Blood flow velocity of MCA did not change after the start of OLV (53.1 +/- 3.2, 55.9 +/- 3.0, 56.4 +/- 2.4, and 54.1 +/- 1.9 vs 50.8 +/- 2.1, 50.7 +/- 2.4, 53.7 +/- 1.5, 50.8 +/- 2.2 cm.sec-1 in groups A and B respectively): there was no difference between the groups. (P < 0.05). CONCLUSION: In clinical practice, the selection of either isoflurane and sevoflurane for OLV was of no difference in terms of the arterial blood oxygenation. With both agents MCA blood flow velocity was maintained during OLV. PMID- 9398956 TI - Prophylactic oral dolasetron mesylate reduces nausea and vomiting after abdominal hysterectomy. The Canadian Dolasetron Study Group. AB - PURPOSE: The incidence of postoperative nausea and vomiting (PONV) varies from 50% to 75% after gynaecological surgery under general anaesthesia. This study evaluates the dose-response relationships, safety, and efficacy of the new 5-HT3 antagonist, dolasetron mesylate, in the prevention of PONV in women undergoing total abdominal hysterectomy (TAH). METHODS: Three hundred and seventy four women scheduled for TAH under general anaesthesia were studied at 13 Canadian centres. Patients received in a randomized, double-blind manner 25, 50, 100, or 200 mg dolasetron or placebo po one to two hours before induction of anaesthesia. The anesthetic protocol was standardized. Efficacy was evaluated for 24 hr after surgery by comparing the number of emetic episodes, administration of rescue medication, severity of nausea, and patient satisfaction. RESULTS: Analysis of complete response (no emetic episodes and no rescue for 24 hr) revealed a linear dose-response relationship across dolasetron groups (P < 0.002). Dolasetron 100 mg (P < 0.003) and 200 mg (P < 0.01) were superior to placebo. The percentage of patients with no emetic episodes increased from 29.3% (placebo) to 54.1 % (100 mg). Subgroup analysis revealed ASA status (I > II), previous history of PONV, previous history of motion sickness, and total morphine dose (> 55 mg associated with less PONV than < 55 mg) influenced the incidence of emetic symptoms, but did not alter the results of the primary analysis. CONCLUSION: Prophylactic dolasetron (100 mg and 200 mg) reduces the incidence of PONV in patients having total abdominal hysterectomy. PMID- 9398957 TI - Peripartum management of a patient with Isaacs' syndrome. AB - PURPOSE: To describe the peripartum management of a patient with Isaacs' syndrome with specific reference to the anaesthetic implications of the disease process. Associated medical problems included obesity, pregnancy induced hypertension and a difficult airway. CLINICAL FEATURES: This 30-yr-old gravida V para 0 woman presented to the anaesthesia consultation clinic at 37-wk gestation to discuss pain relief options for labour and delivery. She had a history of Isaacs' syndrome (a peripheral motor neuron disorder), congenital heart disease (ASD and VSD), treated Hashimotos thyroiditis, obesity and a family history of haemachromatosis. On the day of consultation, she was hypertensive and peripheral oedema was noted. Her urine showed trace protein. Four days later, she presented to the labour suite and her cervix was 9 cm dilated. An epidural anaesthetic was given without difficulty and she had an uneventful labour and delivery course. There were no subsequent neurological complications. CONCLUSION: Isaacs' syndrome is an extremely rare peripheral motor neuron disorder. This patient was successfully managed with epidural analgesia for labour and delivered a healthy child with no congenital anomalies. PMID- 9398958 TI - Spinal artery syndrome masked by postoperative epidural analgesia. AB - PURPOSE: We report a case of a patient who developed a postoperative anterior spinal artery syndrome that was masked by the use of epidural analgesia. We wish to alert other anaesthetists that the use of epidural anaesthesia in this setting may mask the symptoms and delay the diagnosis of this rare complication. CLINICAL FEATURES: The patient was a 22-yr-old obese man with metastatic testicular carcinoma who underwent a left-sided thoracoabdominal retroperitoneal tumour resection. A lumbar epidural catheter was placed preoperatively for pain management. Postoperatively, the patient developed bilateral lower extremity weakness, which was at first attributed to epidural administration of local anaesthetics. Despite discontinuation of the local anaesthetics, the symptoms persisted. Further work-up led to the diagnosis of anterior spinal artery syndrome. The patient was sent to a rehabilitation hospital and had a partial recovery. CONCLUSION: Anterior spinal artery syndrome can occur following retroperitoneal surgery. It is important to recognize the potential for this complication when postoperative epidural analgesia is contemplated, especially following a left-sided surgical dissection. The use of epidural local anaesthetics immediately after surgery delays the diagnosis of a postoperative neurological deficit. Moreover, when the deficit is recognized the epidural itself may be falsely blamed for postoperative paraplegia. If epidural analgesia is used, opioids may be preferred over local anaesthetics in the immediate postoperative period to prevent masking of an anterior spinal artery syndrome. PMID- 9398960 TI - Venous air embolism during orthotopic liver transplantation in a child. AB - PURPOSE: A first case of massive venous air embolism is reported as a complication of orthotopic liver transplantation in a 17-month-old child during the dissection phase. CLINICAL FEATURES: During the hepatic dissection phase, perforation of suprahepatic veins was responsible for an air embolism with a decrease of P(ET)CO2 (27 to 6 mmHg), hypoxaemia (SpO2 decreased from 100 to 88%), and haemodynamic instability (systolic blood pressure decreased from 85 to 50 mmHg, central venous pressure increased from 6 to 10 mmHg). Central venous aspiration of air was unsuccessful but immediate resolution occurred and neurological outcome was normal. CONCLUSION: Venous air embolism during the dissection phase of liver transplantation in children is a risk that should be considered PMID- 9398959 TI - Continuous haemodiafiltration during and after cardiopulmonary bypass in renal failure patients. AB - PURPOSE: Continuous haemodiafiltration (CHDF) is a technique enhancing the efficiency of solute clearance of haemofiltration by infusing dialysis fluid through the haemofilter. It has been reported to control water and electrolyte balance continuously without haemodynamic instability in critically ill patients with renal failure, Therefore, we used CHDF during and after cardiopulmonary bypass (CPB) in two renal failure patients, and discuss its efficacy. CLINICAL FEATURES: The first patient undergoing aortic valve replacement had dialysis dependent renal failure. Chronic renal failure in the second patient undergoing mitral valve replacement and coronary revascularization was controlled preoperatively with diuretics. In both cases, CHDF was performed not only during CPB but also in the post-CPB period. Serum concentrations of potassium, urea and creatinine were well-controlled in spite of large amount of blood transfused in the post-CPB period (1000 ml fresh blood and 400 ml fresh frozen plasma in the fist patient, and 1400 ml fresh blood in the second patient). There was no difficulty in haemostasis during the use of nafamostat mesilate as an anticoagulant to keep activated clotting time at about 150 sec for CHDF in the post-CPB period. CONCLUSION: Our initial experiences of CHDF during and after CPB suggest that the technique provides excellent electrolyte, metabolite and fluid management for the cardiac patients with chronic renal failure. Combined with nafamostat mesilate for anticoagulation, CHDF was simple and safe and did not increase the risk of bleeding. PMID- 9398961 TI - The cost for construction and operation of a simulation centre. AB - PURPOSE: Lack of financial information results in planning difficulties and may delay the introduction of simulator based education. We collected data from an existing simulation centre and describe a construction and operating budget to facilitate planning and construction for interested institutions. METHODS: After obtaining approval from the managing board, the plans and financial statements of the Canadian Simulation Centre, Sunnybrook Health Science Centre, University of Toronto were reviewed from the period from July 1994 through June 1996. Costs were calculated from the financial reports and separated into construction and operation phases. A list of the ongoing educational and research activities was compiled. RESULTS: All dollar figures are expressed in 1996 Canadian Dollars. The planning and construction took place from July 1994 through June 1995. Construction costs for the simulation centre totalled $665,000, of which 85% was related to capital equipment purchases and 15% for salary support. The net costs of ongoing education and research activities (3.35 days/week) were $167,250 from July 1995 through June 1996. About 65% of this consisted of salary support and was absorbed by the existing educational resources of the University of Toronto Department of Anaesthesia. CONCLUSION: Substantial resources are required for the construction of a simulation centre ($665,000) primarily use of capital equipment purchases. However, there is also a considerable operating cost per year ($167,250) which consists mostly of salary support. PMID- 9398963 TI - Assessment of a new ultrafiltration blood processing system. AB - PURPOSE: To report our clinical experience with a new blood processing device which uses ultrafiltration. We assessed safety and efficacy by evaluating: 1 ) the quality and the quantity of intraoperative shed blood processed and reinfused to the patient 2) homologous blood requirements 3) clinical status of the patient post-transfusion. METHODS: With Ethics Committee approval, the ultrafiltration device was used in six consenting patients undergoing major elective spinal surgery. Blood samples for haematology and biochemistry tests were collected from patients post-induction of anaesthesia (baseline), 1 hr and 24 hr post autotransfusion. Volumes of blood collected and processed, and all autologous and homologous transfusions were recorded. Patients were assessed post-operatively for any adverse effects. RESULTS: Five patients had donated blood preoperatively. One patient required homologous blood products in addition to autologous blood. In two patients, the filtration cartridge became blocked and required changing midprocessing. No patient sustained device-related complications. One patient had postoperative haematuria which resolved spontaneously within two hours. CONCLUSION: The ultrafiltration device was safe and effective in reducing homologous blood requirements in six patients undergoing elective spinal surgery. Further evaluation of the ultrafiltration device will be necessary, especially in view of the blockage of the filtration cartridge. PMID- 9398962 TI - Halothane inhibition of acetylcholine-induced relaxation in rat mesenteric artery and aorta. AB - PURPOSE: The effect of halothane was compared on acetylcholine (ACh)-induced relaxation of the mesenteric artery and the aorta in rats. METHODS: The responses of isolated rat aortic and mesenteric arterial ring segments precontracted with phenylephrine to ACh (10(-8)-10(-5) M), in the presence of halothane 0-3%, were compared using isometric force tension recordings. Effects of NG-nitro-l-arginine (L-NOARG, 3 x 10(-5), methylene blue (MB, 5 x 10(-6) M), oxyhaemoglobin in (OxyHB, 10(-7) M), and various potassium channel inhibitors; tetraethylammonium (TEA, 10(-5) M, 10(-3) M), apamin (AP, 10(-7) M), charybdotoxin (ChTx, 10(-7) M) and glibenclamide (GC, 10(-5) M) on ACh-induced relaxation in mesenteric artery were tested. Using radioimmunoassay, ACh (10(-6) M)-induced guanosine 3':5' cyclic monophosphate (cGMP) accumulation of mesenteric arterial rings pretreated with L-NAORG were also measured. RESULTS: L-NOARG partially inhibited ACh-induced relaxation in mesenteric arterial rings (P < 0.05, maximum relaxation reduced by approximately 50%), whereas it abolished them in aortic rings. The remaining relaxation resistant to L-NOARG in mesenteric arterial rings was insensitive to additional MB or OxyHB, and was not accompanied by increases in cGMP contents of rings. Halothane inhibited endothelium-dependent relaxation in aorta and mesenteric arterial rings. This inhibitory effect was larger in aorta. Halothane also inhibited NO independent EDHF-dependent relaxation in the mesenteric arterial rings, CONCLUSION: Despite a similar inhibitory effect on the EDHF relaxing pathway, halothane has a larger effect on endothelium-dependent relaxation in the aorta (NO dependent mainly) than in the mesenteric rings (NO and EDHF dependent). PMID- 9398964 TI - Onset of vecuronium neuromuscular blockade at the hand with an arterio-venous shunt. AB - PURPOSE: To evaluate the onset of vecuronium neuromuscular blockade in the hand with an arterio-venous shunt for haemodialysis. METHODS: In 15 adult patients receiving haemodialysis for renal failure the onset of vecuronium-induced neuromuscular blockade after 0.08 mg-kg-1 vecuronium i.v. was measured. Using train-of-four mechanomyographic monitoring, the force of contraction of the adductor pollicis of both hands with and without arterio-venous shunt was measured simultaneously. RESULTS: The times from the injection to the first depression of twitch response (latent onset) and 95% twitch depression (onset) in the hand with and without arterio-venous shunt were 114.7 +/- 33.4 and 218.7 +/- 59.9 and 117.3 +/- 34.3 and 208.7 +/- 60.9 sec respectively. No difference in the onset of vecuronium neuromuscular blockade in the hand an arterio-venous shunt was demonstrated. CONCLUSION: The presence of an arteriovenous fistula does not modify the onset on neuromuscular blockade. Either arm can be used to monitor onset of neuromuscular blockade in chronic renal failure patients with an arterio venous shunt in the hand for haemodialysis. PMID- 9398965 TI - Prostaglandin E1, lidocaine, and prostaglandin E1-lidocaine combination for attenuating cardiovascular responses to extubation. AB - PURPOSE: Tracheal extubation produces haemodynamic changes that may cause myocardial ischaemia in patients with coronary arterial disease. Intravenous infusion of prostaglandin E1 (PGE1) attenuated the hypertensive response to tracheal extubation but failed to blunt the tachycardia, which was attenuated by intravenous lidocaine. Thus, we investigated whether a combination of PGE1 and lidocaine can overcome the drawbacks of treatment with PGE1 alone. METHODS: One hundred adult patients (ASA 1) undergoing elective minor surgery were randomly assigned to receive one of four treatments: saline (as a control), 1 mg-kg-1 lidocaine, infusion of 0.1 microgram-1.kg-1.min-1 PGE1, or infusion of 0.1 microgram-1.kg-1.min-1 PGE1 plus injection of 1 mg-1.kg-1 lidocaine. Lidocaine was injected two minutes before tracheal extubation. The PGE1 was infused from completion of surgery until five minutes after tracheal extubation. Anaesthesia was maintained with sevoflurane 1.0%-2.5% and nitrous oxide 60%. Heart rate (HR) and blood pressure (BP) were measured before and after tracheal extubation. RESULTS: Lidocaine alone and PGE1-lidocaine combination attenuated the increases in BP and HR observed in the control group: PGE1 alone was effective in attenuating hypertensive response but ineffective for tachycardia. The suppressive effect of the PGE1-lidocaine combination on BP increase was superior to that of each drug alone, and the combined effect on HR increase was similar to that of lidocaine alone. CONCLUSION: The combination of PGE1 infusion and lidocaine is a more effective method of attenuating hypertension and tachycardia associated with tracheal extubation than either drug alone. PMID- 9398966 TI - Canadian Anaesthetists' Society Gold Medal. PMID- 9398967 TI - Inhalation induction with sevoflurane. PMID- 9398968 TI - Cricoid pressure. PMID- 9398969 TI - Stethoscopy. PMID- 9398970 TI - Anaesthesia and congenital tracheal stenosis. PMID- 9398971 TI - Bias in uncontrolled brain tumor trials. PMID- 9398972 TI - The crisis of biomedical research funding in Canada. PMID- 9398973 TI - Neurotrophin regulation of gene expression. AB - The neurotrophins comprise a family of secreted proteins that elicit profound responses in cells of the developing and mature vertebrate nervous system including the regulation of neuronal survival and differentiation. The molecular mechanisms by which the neurotrophins exert their effects have been the subject of intense investigation. The neurotrophins elicit responses in neurons via members of the Trk family of receptors and the p75 neurotrophin receptor. Once activated, neurotrophin receptors trigger a large number of biochemical events that propagate the neurotrophin signal from the plasma membrane to the interior of the cell. An important target of the neurotrophin-induced signaling pathways is the nucleus, where neurotrophin-induced signals are coupled to alterations in gene expression. These neurotrophin-induced changes in gene expression are critical for many of the phenotypic effects of neurotrophins including the regulation of neuronal survival and differentiation. PMID- 9398974 TI - Role of the ipsilateral motor cortex in voluntary movement. AB - The ipsilateral primary motor cortex (M1) plays a role in voluntary movement. In our studies, we used repetitive transcranial magnetic stimulation (rTMS) to study the effects of transient disruption of the ipsilateral M1 on the performance of finger sequences in right-handed normal subjects. Stimulation of the M1 ipsilateral to the movement induced timing errors in both simple and complex sequences performed with either hand, but with complex sequences, the effects were more pronounced with the left-sided stimulation. Recent studies in both animals and humans have confirmed the traditional view that ipsilateral projections from M1 to the upper limb are mainly directed to truncal and proximal muscles, with little evidence for direct connections to distal muscles. The ipsilateral motor pathway appears to be an important mechanism for functional recovery after focal brain injury during infancy, but its role in functional recovery for older children and adults has not yet been clearly demonstrated. There is increasing evidence from studies using different methodologies such as rTMS, functional imaging and movement-related cortical potentials, that M1 is involved in ipsilateral hand movements, with greater involvement in more complex tasks and the left hemisphere playing a greater role than the right. PMID- 9398975 TI - Neural transplantation in Parkinson's disease. AB - Parkinson's disease is a neurodegenerative disorder that affects about 1% of Canadians between the ages of fifty and seventy. The medical management for these patients consists of drug therapy that is initially effective but has limited long term benefits and does not alter the progressive course of the disease. The recalcitrance of longstanding Parkinson's disease to medical management has prompted the use of alternative surgical therapies. Many neurosurgical procedures have been utilized in order to improve the disabling symptoms these patients harbour. Although most of the current procedures involve making destructive lesions within various basal ganglia nuclei, neural transplantation attempts to reconstitute the normal nigrostriatal pathway and restore striatal dopamine. The initial success of neural transplantation in the rodent and primate parkinsonian models has led to its clinical application in the treatment of parkinsonian patients. Currently, well over one hundred patients throughout the world have been grafted with fetal tissue in an effort to ameliorate their parkinsonian symptoms. Although the results of neural transplantation in clinical trials are promising, a number of issues need to be resolved before this technology can become a standard treatment option. This review focuses on the current status of neural transplantation in Parkinson's disease within the context of other surgical therapies in current use. PMID- 9398976 TI - Hormones, radiosurgery and virtual reality: new aspects of meningioma management. AB - The understanding and management of meningiomas is changing significantly today. One of the most striking features of their pathophysiology is their predominance in women. In a series of 517 patients with meningiomas seen by the Brain Tumor Group at Brigham and Women's Hospital, the female:male ratio was 24:1. The progesterone receptor appears to be the major candidate to explain this difference. Although meningioma cells variably express receptors for estrogen, androgen, platelet-derived growth factor, epidermal growth factor, and somatostatin, these molecules do not explain the differences because they are not differentially expressed or are not activated. Progesterone receptor can be shown to be expressed in 81% of women and 40% of men with meningiomas; it can also be shown to be activated by transfecting a construct with the progesterone responsive element and a reporter in it and using the cell's own receptors to activate this construct. Surgery remains the mainstay of meningioma management. At the Brigham and Women's Hospital three-dimensional reconstruction techniques have markedly improved the ability to visualize the tumor as well as its relation to vascular structures. With MRI reconstruction, it is possible to know the tumor's relation to the sagittal and other sinuses, to identify feeders and proximity to major arteries, and to establish its location and relation to cortex by frameless stereotaxis. These techniques can be used in a virtual reality format are some of the most powerful in neurosurgery both for teaching and for the surgical procedure itself. External beam radiation has been shown by others to be an effective adjunctive treatment to prevent meningioma recurrence. Recently, linear accelerator radiosurgery and stereotactic radiotherapy have changed the pattern of radiation at our institution. In a series of 56 skull base meningiomas, for example, 95% were controlled (i.e., showed no growth) over a four year period. Fractionated focal radiation potentially offers the same control rate with fewer complications. With increasing understanding and treatment possibilities, meningiomas remain one of the most intriguing and challenging tumors in the nervous system. PMID- 9398977 TI - Measuring bias in uncontrolled brain tumor trials--to randomize or not to randomize? AB - PURPOSE: To help investigators decide if new therapies for glioma warrant definitive evaluation in randomized studies we have been developing a method for assessing the degree to which patient selection may have enhanced the results of uncontrolled treatment trials. In this study, we analyzed the impact of case selection on the survival of patients with malignant glioma receiving adjuvant stereotactic radiosurgery, a promising therapy reserved for those with small tumors and good performance status. METHODS: Following published eligibility criteria we simulated the patient selection process for stereotactic radiosurgery given as a boost at the conclusion of conventional radiotherapy. Eligible patients were culled from a pre-existing clinical/imaging database of 101 consecutive conventionally-treated patients with biopsy-proven malignant glioma and known survival times. Median durations of survival and 2- and 3-year survival rates were determined for those judged eligible or ineligible for stereotactic radiosurgery. RESULTS: Twenty-seven percent of patients were deemed eligible for stereotactic radiosurgery, eligible patients had more favorable prognostic factors and significantly longer median survival than ineligible patients (23.4 vs. 8.6 months; 2-year rate, 48% vs. 15%; 3-year rate, 30% vs. 7%); eligible patients also had a longer median survival than the entire group of unselected patients (23.4 vs. 11.4 months). Radiosurgery-eligible, conventionally-treated patients with glioblastoma multiforme and a group of radiosurgery-treated patients at a special referral center had similar median survival times (16.4 vs. 19.7 months). CONCLUSION: We provide additional evidence for selection bias in uncontrolled trials of stereotactic radiosurgery and by simulating the selection process accurately have detected a larger bias effect than noted previously. Judging from experience with interstitial radiation and intraarterial chemotherapy where substantial selection bias also occurred and randomized controlled trials proved disappointing, we conclude that a phase III study of stereotactic radiosurgery for malignant glioma is unlikely to yield a positive result and may not be necessary. PMID- 9398978 TI - Prognostic factors in oligodendroglioma. AB - BACKGROUND: A reliable marker for tumor oligodendroglial cells is not yet available, so that the histological recognition of the tumor still encounters uncertainties. There is no general agreement also on prognostic factors in oligodendroglioma. The inconsistency concerns mainly the histopathological factors. The aim of the study was recognition of prognostic factors in oligodendroglioma. METHODS: In a series of ninety-eight oligodendrogliomas, including twenty mixed oligoastrocytomas, clinical [sex, age at surgery, tumor location, symptoms at presentation], therapeutic [extent of resection, year of surgery, post-operative Karnofsky score, post-operative radiotherapy, post operative chemotherapy], histological [cell density, nuclear pleomorphism, vascular endothelial proliferation, necrosis, microcysts, mitoses, mitotic index (MI), apoptosis, apoptotic index (AI)] and immunohistochemical parameters [MIB-1 and PCNA Labeling Indexes (LIs), staining for GFAP, positivity for p53] were correlated with survival in uni- and multivariate analysis in order to identify their prognostic significance. RESULTS: Age at surgery, extent of surgical resection, year of surgery, post-operative Karnofsky score and MIB-1 LI were associated with survival in both uni- and multivariate analysis. Location, symptoms at presentation, mitoses, MI, AI, and PCNA LI showed a significant correlation with survival in uni- but not in multivariate analysis. The twenty cases of oligoastrocytomas did not show any difference in survival from pure oligodendrogliomas. CONCLUSIONS: Some clinical and therapeutic factors together with MIB-1 LI play a prognostic role. MIB-1 LI is prognostic with a cutoff of 8%. Histology gives a limited contribution to the prognosis. Oligoastrocytomas had the same outcome and prognostic factors as pure oligodendrogliomas. PMID- 9398979 TI - An electroencephalographic classification for coma. AB - BACKGROUND: The assessment of thalamocortical function in comatose patients in the intensive care unit (ICU) can be difficult to determine. Since the electroencephalogram (EEG) affords such assessment, we have developed an EEG classification for comatose patients in our general ICU. METHODS: One hundred EEGs were classified in a blinded fashion by two EEGers, using our method and that of Synek. Interobserver agreement was assessed using kappa score determination. RESULTS: Kappa scores were 0.90 for our system and 0.75 for the Synek system. (The Kappa score represents the inter-rater agreement that is beyond chance; 0.90 is almost perfect agreement, while 0.75 is substantial agreement). CONCLUSION: Our system for classifying EEGs in comatose patients has a higher interobserver reliability than one that was previously published. This EEG classification scheme should be useful in clinical electrophysiological research involving ICU patients, allowing for internal consistency and comparisons among centres. PMID- 9398980 TI - Familial intracranial aneurysms: recurrence risk and accidental aggregation study. AB - BACKGROUND: The Saguenay-Lac-Saint-Jean (SLSJ) region is a geographically isolated area (population 285,955) located in the Northeastern part of the Province of Quebec, Canada. Using a population-based register, the genealogical reconstruction of 502 individuals with ruptured intracranial aneurysm (RIA) showed a familial aggregation (the presence of aneurysm in two or more first- to third-degree relatives) for 144 (28.7%) of them; this proportion is much higher than reported elsewhere. OBJECTIVE: In order to assess the genetic predisposition to RIA in the SLSJ population, the objective of the present study is to compare familial and non-familial cases and to provide an estimate of the recurrence risk ratio for siblings. RESULTS: The age at the time of rupture, the number of intracranial aneurysms for each patient and the location of RIAs were not statistically different in the familial versus the non-familial group. Of the 3449 siblings, 20 (0.58%) had suffered a RIA. The recurrence risk ratio calculated for siblings (defined as the risk of disease among siblings divided by the estimated population prevalence) is 1.6 (CI 95% 1.0-2.4). In other respects, we observed very large kinships in the SLSJ population, with an average number of siblings of 7.2 (SD +/- 3.4), ranging from 0 to 17 individuals. With such large families and on the basis of chance alone, we expected 31.3% of the patients to have at least one first- to third-degree relative with RIA. CONCLUSION: These data show that siblings of patients with RIA in the SLSJ population have a greater risk of RIA than the general population. Nevertheless, the largest part of the familial occurrence observed in the SLSJ region can be explained by accidental aggregation, due to large kinships. We propose that, in this population, an underlying genetic predisposition must be suspected only when three or more cases of RIA are identified among first- to third-degree relatives. PMID- 9398981 TI - Motor evoked potentials and disability in secondary progressive multiple sclerosis. AB - BACKGROUND: To investigate the mechanisms underlying disability in multiple sclerosis (MS), 40 patients with the relapsing-remitting form of the disease and 13 patients with secondary progressive MS underwent multimodal evoked potential (EP), motor evoked potential (MEP), and spinal motor conduction time evaluation. Clinical disability was evaluated by the expanded disability status scale (EDSS) and functional system scales. In secondary progressive MS patients, magnetic resonance imaging (MRI) was used to obtain a semiquantitATive estimate of the total lesion load of the brain. RESULTS: Spinal motor conduction time was significantly longer in secondary progressive MS patients than controls (p < 0.001) and relapsing-remitting MS patients (p < 0.05), but did not differ between relapsing-remitting patients and controls. Spinal motor conduction times also correlated directly with EDSS scores (p < 0.001) and pyramidal functional system scores (p < 0.001). Brain lesion load (4960.3 +/- 3719.0 mm2) and the total number of lesions (67.7 +/- 37.0) in secondary progressive MS did not correlate with disability scores. For the following EPs, the frequencies of abnormalities were significantly higher in secondary progressive MS patients than relapsing remitting patients: visual evoked potentials (p < 0.05), somatosensory evoked potentials and upper limb motor evoked potentials (p < 0.01), and brainstem auditory evoked potentials, lower limb somatosensory evoked potentials and lower limb motor evoked potentials (p < 0.001). CONCLUSIONS: These findings suggest that disability in secondary progressive MS patients is mainly due to progressive involvement of corticospinal tract in the spinal cord. PMID- 9398982 TI - Symptoms experienced by patients with carpal tunnel syndrome. AB - BACKGROUND: Patients with carpal tunnel syndrome (CTS) sometimes report sensory symptoms outside the median nerve distribution. This study was designed to provide a more detailed assessment of these symptoms. METHODS: Patients with clinical suspicion of upper limb neuromuscular lesions were divided into those with electrodiagnostic (EDX) evidence of CTS, and those without. CTS patients with superimposed nerve abnormalities were excluded. Motor and sensory symptoms were assessed in the exclusive CTS patients. RESULTS: Over 50% of patients with exclusive CTS reported tingling or numbness over the whole hand, ulnar or radial nerve distributions. Some patients reported symptoms proximal to the wrist. Sensory signs did not extend beyond the median nerve distribution. Numbness and nocturnal pain were predictive of positive EDX evidence of CTS. CONCLUSIONS: Sensory symptoms outside the distribution of the median nerve are common in CTS. For enhanced sensitivity in diagnosis it is useful to be aware of these "atypical" symptoms. Reports of numbness and nocturnal pain are strong indicators of CTS. PMID- 9398983 TI - Reversal of syphilitic hydrocephalus with intravenous penicillin. AB - BACKGROUND: CSF shunting procedures are generally considered the fundamental therapy of syphilitic hydrocephalus. METHODS: We followed up with CSF analysis and MR imaging a patient with progressive mental and gait disturbances and tetraventricular hydrocephalus due to tertiary syphilis who was treated for 14 days with high dose intravenous penicillin alone. RESULTS: Clinical and CSF abnormalities resolved within a few months, whereas the hydrocephalus disappeared only 30 months after therapy. CONCLUSIONS: Before consideration of a CSF shunting procedure, a trial of high dose intravenous penicillin is warranted for patients with syphilitic hydrocephalus. PMID- 9398984 TI - Epilepsy and driving: a survey of Canadian neurologists. AB - BACKGROUND: A seizure is the most common cause of loss of driving privileges for medical reasons but there is variability in how physicians and the authorities who regulate driving approach this issue. METHODS: A questionnaire regarding epilepsy and driving was sent to all adult neurologists in Canada (n = 494). RESULTS: Of 289 (59%) neurologists responding, 50% usually report patients with seizures to the department of motor vehicles compared to only 4% for stroke/TIA, 26% for dementia and 8% for other neurologic disorders (p < 0.0001). In the five provinces with mandatory reporting laws, seizures were reported most of the time by 84% compared to only 19% in the five provinces with discretionary reporting (p < 0.0001). Nationwide, 44% agreed with mandatory reporting but this also differed in provinces with and without mandatory reporting legislation (63% vs. 37%, p < 0.0001). Only 49% agreed with the current recommendation of at least one year seizure free interval before resuming driving. CONCLUSIONS: Seizures are disproportionately reported compared to other neurological conditions. Many neurologists disagree with the recommended Canadian standards for duration of driving restriction after seizures. Variability in the attitude and practice of neurologists in regard to reporting of seizures is confirmed. PMID- 9398985 TI - What effect does an untreated aneurysm have on life expectancy. PMID- 9398986 TI - Early seizures after closed head injury. PMID- 9398987 TI - Musical hallucinosis with brain lesions. PMID- 9398988 TI - Comparative study of invasive squamous cell carcinoma and verrucous carcinoma of the oral cavity: expression of bcl-2, p53, and Her-2/neu, and indexes of cell turnover. AB - Significant clinical and biological differences are found between invasive squamous cell carcinoma (SCCA) and verrucous carcinoma (VC) of the oral cavity. The correct diagnosis of these tumors has important therapeutic implications. Immunoperoxidase stains for bcl-2, p53, and Her-2/neu, and in situ end-labeling of DNA to identify apoptosis were performed in eight VC and eight SCCA matched for age, sex, and stage. Marked differences were identified in the pattern of expression of oncogenes and the indexes of cell turnover in these two types of tumors. VC displayed minimal apoptosis in rare keratinizing cells (0 to 3%); p53 positive cells (4/8) and Ki-67 (8/8) were confined to the nuclei of the basal proliferating layers; and bcl-2 (4/8) was expressed only in the cytoplasm of rare tumor cells. In contrast, SCCA displayed higher apoptosis rates (5 to 10%), whereas p53- (5/8) and Ki-67- (8/8) positive nuclei were distributed randomly throughout the tumor. Very well differentiated areas in one SCCA case had a pattern of staining for p53 and Ki-67 similar to the one seen in VC. In SCCA bcl 2 showed patchy cytoplasmic staining (4/8) or strong cytoplasmic and nuclear positivity (2/8) in the less differentiated tumors. Her-2/neu was negative in all VC and SCCA cases. The different levels and patterns of gene expression and cell turnover between SCCA and VC undoubtedly correlate with the different biology and prognosis of these tumors. PMID- 9398989 TI - Rapid diagnostic imaging of cancer using radiolabeled liposomes. AB - A novel tumor diagnostic imaging method was developed that allows tumor localization soon after administration of radiolabeled liposomes. Although previous studies showed that radiolabeled liposomes can reach various tumors in a short time, their blood clearance is slow, and the high blood background hinders early imaging. Therefore, we attempted to remove actively the liposomes from the circulation using the strong affinity between avidin and biotin. Liposomes that had biotin bound to their surface and were labeled with 111In, 67Ga, or 99mTc were administered to mice bearing sarcoma 180, followed by administration of avidin 2 or 4 h later. Avidin initiated liposomal aggregation, resulting in their rapid removal by the reticuloendothelial system. Consequently, their blood level was markedly reduced without any changes in tumor levels. The tumor-to-blood ratio reached about 13 at only 2.5 h after administration of 99mTc-labeled liposomes, versus 1.0 or less without postadministration of avidin. Increased liver accumulation was also observed, but it decreased gradually with time. PMID- 9398990 TI - Smoking history and cancer patient survival: a hospital cancer registry study. AB - While tobacco use is clearly the most preventable cause of cancer, little is known about whether smoking adversely influences cancer patients' survival. The goal of this study was to examine the effect of smoking history on survival among cancer patients. Data from Memorial Sloan-Kettering Cancer Center's tumor registry was used to identify 25,436 cases of cancer (12,447 male patients and 12,989 female patients). Information regarding smoking and alcohol consumption, histologic grade, tumor stage, and survival time was available. Proportional hazard analysis was used to examine the effect of smoking on the death from all causes among patients. Patients who had a history of smoking were found to have a lower rate of survival than nonsmokers. After controlling for age, race, alcohol use, and histologic grade, the risk ratios were 1.55 for males and 1.43 for females. A dose-response relationship was found between ever-smoking and cancer patient survival. The predictive effect of smoking on survival was significant for patients with oral, pancreatic, breast, and prostate cancers, but not for esophageal, stomach, colon, rectum, laryngeal, lung, cervix uteri, urinary bladder, and kidney cancers. Black patients with oral or breast cancer had a poorer prognosis associated with smoking compared with white and other nonwhite patients. The strongest effect of smoking on survival was found mainly among patients with breast cancer with a distant tumor stage or with prostate cancer with a regional tumor stage. Alcohol use alone was associated with a higher risk of death for nonsmokers with oral and pancreatic cancers than for similar cancer patients without a history of alcohol use. Smoking history plays a critical role in influencing cancer patient survival, especially for patients diagnosed with oral, pancreatic, breast, or prostatic cancers. In addition, alcohol consumption is independently related to survival in patients with oral and pancreatic cancers. Our study suggests that a potential means of improving cancer patient survival, especially from oral, pancreatic, breast, and prostatic cancers, may be achieved through smoking cessation. PMID- 9398991 TI - Factors associated with breast and cervical cancer screening practices among Vietnamese American women. AB - PURPOSE: To investigate predictors of breast and cervical cancer screening tests among Vietnamese women in California in preparation for developing and testing interventions to promote such screening. METHODS: Cross-sectional telephone survey of 933 randomly selected Vietnamese women in four California counties. RESULTS: Overall, 70% of the respondents had had at least one prior clinical breast examination, but only 30% had had a mammogram and 53% a Pap test. Among women who had been screened, more than two-thirds were up-to-date and among those who had not been screened, more than two-thirds were planning future tests. Factors positively associated with receipt of one or more of the tests included age (among women < 40 years old), number of years in the United States, having ever married, and having health insurance. Factors negatively associated with test receipt included having a Vietnamese doctor, being unemployed, and being of Chinese-Vietnamese background. CONCLUSION: The multiple factors associated with utilization suggest intervention targets for promoting breast and cervical screening among new immigrant women. Increasing screening test receipt to recommended levels will require a two-pronged approach directed at both Vietnamese consumers and Vietnamese physicians. PMID- 9398992 TI - Tamoxifen and ocular toxicity. AB - Tamoxifen, widely used in the management of breast cancer, has been associated with a reduction of mortality and recurrence as well as occurrence of controlateral tumors. It is generally well tolerated, apart from certain well documented adverse effects concerning mainly the reproductive organs, the most worrying being its carcinogenicity for the endometrium. Ocular toxicity has also been reported as one possible complication of the drug, with lesions described in the retina, the cornea, or the optic nerve, especially in women treated with high daily or cumulated doses of tamoxifen, although some cases have also been reported with standard doses. The incidence of such ocular complications is rather low considering the large number of patients receiving tamoxifen. The possible reversibility of these lesions, if discovered in time, emphasizes the need for clinicians to be aware of these ocular reactions and raises the question of periodic ophthalmological screening examinations among patients receiving tamoxifen. PMID- 9398993 TI - Immunohistochemical assessment of proliferation markers and altered gene expression in archival specimens of ovarian epithelial tumors. AB - Recently reported morphologic and molecular genetic evidence suggests that some ovarian carcinomas arise from their benign and low malignant potential (LMP) counterparts. In order to help reach a better understanding of ovarian tumorigenesis, we studied a wide range of gene products involved in cellular growth regulation in archival material obtained from three groups of tumors with graduated malignant potential. Immunohistochemical staining was performed for Ki 67, proliferating cell nuclear antigen (PCNA), epidermal growth factor receptor (EGFR), HER-2/neu-encoded receptor protein, p53 gene product, and multidrug resistance gene product (P-glycoprotein). The expression of EGFR, HER-2/neu encoded receptor protein, and mutant p53 product was significantly lower in LMP tumors than in carcinomas (p < 0.05). HER-2/neu immunopositivity was more prevalent in adenocarcinomas than in LMP tumors, and the proportion of HER-2/neu positive adenocarcinomas increased with the progression of the disease. The staining differences between LMP tumors and adenocarcinomas with antibodies against Ki-67, PCNA, and P-glycoprotein were not statistically significant. Immunohistochemical detection of EGFR, HER-2/neu, and p53 in ovarian epithelial tumor is relevant to ovarian tumorigenesis. It could serve as a powerful tool for the pursuit of retrospective studies focused on these important biologic markers. PMID- 9398994 TI - Deficient DNA repair in chronic ulcerative colitis. AB - Carcinoma of the colon is a serious complication of chronic ulcerative colitis (CUC), a disease of unknown etiology. Peripheral blood lymphocytes from nine patients with CUC showed deficient repair of radiation-induced DNA damage compared with a group of healthy controls. DNA repair was measured indirectly by quantifying chromatid breaks after irradiation of cells with X-rays or ultraviolet during G2 phase of the cell cycle. Such breaks represent unrepaired DNA strand breaks that may arise directly from the damaging agent or indirectly during repair processes. Two types of deficiency were revealed. One was an abnormally high frequency of chromatid breaks after G2-phase X-irradiation. These may reflect deficient strand-break repair. The second deficiency was manifest as a low frequency of breaks not increased by addition of the DNA repair inhibitor araC. This low frequency apparently results from negligible incision activity. Deficient DNA repair in CUC may thus be a requisite predisposing factor for genomic instability and the potential development of colon carcinoma. PMID- 9398995 TI - Pancreatic cancer cell regulation by lipids and by basic fibroblast growth factor expression. AB - High fat intake is a risk factor for pancreatic cancer. Lipids may act either directly or in cooperation with growth-promoting polypeptides. In this study, the role of serum lipids, and mainly the often expressed intracellular basic fibroblast growth factor (bFGF) isoforms in cancer cells, was analyzed in pancreatic tumor cell proliferation. Serum lipids alone induced a 1.9-fold increase of human pancreatic cancer cell growth (p < 0.001). Treatment with bFGF had a weak mitogenic effect (1.2- to 1.3-fold increase) compared with those of insulin and transferrin (1.7- to 1.6-fold increase, respectively). The bFGF expression by a rat pancreatic cancer cell line that was transfected with bFGF cDNAs modified cell lipid contents and induced a higher proliferation rate than that found with the exogenous bFGF. Combined extra- and intracellular bFGFs increased cell growth by two to three times (p < 0.001), regardless the presence of extracellular lipids. The results obtained reflect the direct mitogenic effect of serum lipids and suggest that the endogenous bFGF of high molecular weight may be implicated in pancreatic cancer cell growth. By modifying cell lipids, bFGFs may interfere with other cell functions, like signal transduction. PMID- 9398996 TI - Effects of catechols on free radical formation by chemotherapeutic agents (adriamycin, farmorubicin, and mitomycin). AB - The aim of the present study was to investigate the effects of biologically important catechols on the cytotoxicity of adriamycin, farmorubicin, and mitomycin C with respect to hydroxyl radical production. Catecholamines (adrenalin, noradrenaline, dopamine) and DOPA enhance the generation of hydroxyl radicals by chemotherapeutic antibiotics. Measures were done using a deoxyribose assay, in presence of the Co(II) + H2O2 system. Catalase and hydroxyl radical scavengers (mannitol, thiourea, cysteine, glutathione, L-lactic dehydrogenase) inhibited the deoxyribose damage caused by the drugs. PMID- 9398997 TI - Visual disturbances in migraine. PMID- 9398998 TI - The rise and fall of estrogen levels. PMID- 9398999 TI - A negative clinical study in the search for a migraine treatment. PMID- 9399000 TI - Role of the suprachiasmatic nucleus in the pathogenesis of migraine attacks. AB - Neuroanatomic, morphometric, immunocytochemical, neurobiochemical and clinical data support the hypothesis that the suprachiasmatic nucleus of the hypothalamus might be the initial site of migraine attacks. The prodromal phase of a migraine attack could be considered a syndrome of functional suprachiasmatic nucleus insufficiency, and other phases a reactive denervation hypersensitivity with the affection of the visual, nociceptive, antinociceptive and cranial vasomotor system. PMID- 9399001 TI - Olfaction in migraine. AB - Olfactory thresholds for acetone and vanillin and the unpleasantness rating of concentrated acetone were measured in 20 migraine sufferers and 21 controls. The olfactory threshold for vanillin was lower in migraine sufferers than in controls. In addition, patients who reported that odours frequently seemed stronger during attacks of migraine were able to detect acetone at a lower concentration than most other patients. No differences were found between migraine sufferers and controls for ratings of the unpleasantness of concentrated acetone. These findings suggest that hyperacuity to odours persists between episodes of migraine. Sensitivity to odours could contribute to the migraine predisposition. PMID- 9399002 TI - Light-induced discomfort and pain in migraine. AB - Quantitative thresholds for discomfort and pain with monocular and binocular light stimuli were measured in 67 controls and 67 migraine patients (37 migraine with aura and 30 migraine without aura). Patients were more photophobic during attack than outside attack (p < 0.03), and they were more sensitive to light than controls even between attacks (p < or = 0.0001). We found no differences in light sensitivity between migraine with aura and migraine without aura (p > or = 0.93). Unilateral pain affected light sensitivity on both sides. When asked with a questionnaire, 74% of patients answered that they were sensitive to light outside attack and 100% were sensitive during attack. Pain thresholds were generally lower among sensitive than non-sensitive patients (p = 0.004), indicating some agreement between subjective opinion and objective measurements of photophobia. Photophobia seems to be an intrinsic property of migraineurs. It is increased by migraine pain, but seems to be unrelated to migraine characteristics such as nausea, severity of attacks, pain character and pain laterality. PMID- 9399003 TI - Simultaneous recording of pattern reversal electroretinograms and visual evoked potentials in migraine. AB - We recorded full-field pattern reversal electroretinograms (PERGs) and visual evoked potentials (PVEPs) simultaneously in 15 migraine with aura, 14 migraine without aura patients during the interictal period, and in 23 sex- and age matched normal subjects. All subjects had normal visual fields. The visual aura in all patients was hemianopsia or fortification spectra. Neither migraine group showed significant differences from normal in latency and amplitude of PERGs. In migraine with aura, the amplitudes of PVEPs in classic migraine at the mid occipital electrode were significantly (p < 0.01) higher than normal. PVEP amplitudes were significantly (p < 0.01) higher on the contralateral side of the aura than the ipsilateral side in both visual aura and normal subjects, but there was no significant difference in latency. This high amplitude and asymmetry of PVEPs may contribute to defective inhibition between interhemispheric visual occipital areas or striate and peristriate areas. PMID- 9399004 TI - The reproducibility of cephalic pain pressure thresholds in control subjects and headache patients. AB - Pain pressure thresholds (PPT) were measured at 13 cephalic points bilaterally in 30 headache patients (10 with tension-type headache, 10 with migraine and 10 with cervicogenic headache) and 10 control subjects on three different days. During the sessions, the subjects reported their pain intensity on the right and left sides of the head on a visual analogue scale (VAS). The variability between days was estimated as a coefficient of repeatability (CR = 2 standard deviations of intraindividual differences). The mean CR across all 13 locations was larger in headache patients (2.0 kg/cm2) than in controls (1.68 kg/cm2). Variability (CR) was larger in headache patients as compared to control subjects for 11 of the 13 points (p = 0.02). Reliability was better in controls (intraclass correlation coefficients (ICC) ranging from 0.55 to 0.85) than in headache patients (ICC ranging from 0.43 to 0.78). A moderate negative association between PPT and pain intensity was demonstrated. The intraindividual PPT difference (PPT on the most painful occasions-PPT on the least painful occasions) was negative at 12 of 13 cephalic points (p = 0.003, across-location mean difference: -0.20 kg/cm2). The PPT differed significantly from one day to the next. A part of this variation was presumably related to the circumstances around the procedure; thresholds were lower when the subjects came directly to algometry without any preceding medical examination at all 13 points (p = 0.0002). These results have implications for the planning of future algometer studies. The sample size that is required in studies of headache patients is greater than that in studies of healthy subjects, especially when patients suffer from pain during the PPT session. Particular attention should be paid to circumstances (e.g. preceding medical investigations) around the algometry procedure in order to reduce variability. PMID- 9399005 TI - Power spectral analysis of heart rate and diastolic blood pressure variability in migraine with and without aura. AB - Autonomic function in migraineurs during headache-free periods was studied by means of cardiovascular reflexes and power spectral analysis of heart rate and diastolic blood pressure variability. We examined 56 patients: 37 suffering from migraine without aura and 19 from migraine with aura. Cardiovascular responses to the tilt test and Valsalva manoeuvre showed a normal function of the overall baroreceptor reflex arc. Normal heart rate responses to Valsalva manoeuvre and deep breathing suggested an intact parasympathetic function. Power spectral analysis of both heart rate and diastolic blood pressure variability in basal conditions and during orthostatic test showed similar sympathovagal interactions modulating cardiovascular control in migraine patients and in controls. PMID- 9399006 TI - Platelet sulphotransferase activity, plasma sulphate levels and sulphation capacity in patients with migraine and tension headache. AB - Activity of both the M- and P-forms of sulphotransferase (ST) was measured in platelets from patients with migraine, tension headache and controls. Mean PST values were 0.065 +/- 0.023 and 0.057 +/- 0.052 nmol/mg protein/min for migraine patients with and without aura. The corresponding values for tension headache and controls were 0.122 +/- 0.059 and 0.127 +/- 0.093 nmol/mg protein/min respectively (p < 0.05). Mean MST values were not different for any of the groups, and MST and PST activities measured in two patients during a migraine attack were not significantly altered from baseline levels. Mean plasma inorganic sulphate concentrations and paracetamol metabolites were not significantly different in any of the groups studied. The results suggest that PST activity may be a factor in the aetiology of migraine. PMID- 9399007 TI - Headache during pregnancy. AB - A questionnaire was submitted to 430 women 3 days after delivery, asking mainly about features of headache before and during pregnancy, and their possible modification or recurrence; moreover, delivery modalities and the condition of the newborn were evaluated. One-hundred-and-twenty-six (29.3%) were found to be primary headache sufferers (IHS criteria, 1988), 81 of whom had migraine without aura (MO), 12 migraine with aura (MA), and 33 tension-type headache (TH). In all three groups, about 80% showed complete remission or a higher than 50% decrease in the number of attacks. The improvement was more evident after the end of the first trimester; this trend was common to the three primary headaches considered. In our series of primary headaches, there was only one case (MO) which began during pregnancy. In a subgroup of pluripara, headache maintained the improvement presented in the first pregnancy also during the following gravidic periods in about 50% of cases, whereas in the remaining 50% a worsening in parallel with successive pregnancies was found. Primary headaches "per se" do not seem to increase the pregnancy or delivery risks, nor the vitality of the newborn. During pregnancy, drug use was very much reduced and was restricted to a limited number of compounds. PMID- 9399008 TI - Epidemiology of migraine headache in Santiago, Chile: a prevalence study. AB - OBJECTIVE: To describe the importance of migraine in Santiago, Chile, by analyzing its prevalence, clinical features and impact by age, gender and socioeconomic status. METHODS: In 1993, a representative sample of 1,540 adults of the province of Santiago were interviewed using a standard questionnaire. A total of 1,385 (89.9%) subjects responded to the survey. Initially, a designated member of each household responded to the questionnaire. Subsequently, each household member with headaches was asked to respond to questions about severity, frequency, location, duration, associated symptoms and impact in work and social activities of their most frequent headaches. Migraine diagnoses were determined in accordance with the International Headache Society (IHS) criteria of 1988. RESULTS: Recurrent headaches in the past year were found in 516 (36.82%) respondents, 145 (28.1%) males and 371 (71.9%) females. Total prevalence of migraine was found to be 7.3% (95% CI 5.9-8.6); 11.9% (95% CI 9.6-14.2) in females and 2.0% (95% CI 0.9-3.0) in males. Overall, migraine constituted 19.6% (101/516) of all headaches reported in this sample. The prevalence did not vary significantly by age groups or socioeconomic status (SES). Migraine with aura had an overall prevalence of 3.5% (CI 0.8-7.1), and was significantly more frequent in females. In 60-70% of cases the attacks lasted 2-6 h and the frequency was 3.3 and 3.4 per month in females and males respectively. Both males and females reported significantly high percentages of attacks during work. CONCLUSIONS: Migraine prevalence in a sample of adults of Santiago is similar to that reported in previous studies using IHS criteria. Women of all socioeconomic levels are at an increased risk. PMID- 9399009 TI - Post-lumbar puncture headache: clinical features and suggestions for diagnostic criteria. AB - The aim of the present prospective study was to describe clinical features of post-lumbar puncture headache (PPH), and to test the validity of the diagnostic criteria of the International Headache Society (IHS). Eighty-eight of the 239 included patients (36.8%) experienced PPH. Females were affected more frequently than males (45.2% vs 21.4%; p < 0.001). First onset of PPH occurred within the first day in 40 patients (53%), within 2 days in 89%, and never after the fourth day. When PPH occurred for the first time on the day the lumbar puncture was performed, it was usually experienced much later in the day (median 14.00 h) than it first occurred on the second day (median 09.30 h) or later. The median duration of PPH was 6 days (range 1-29 days). Patients with headache performed a "Rising Manoeuvre" twice daily as long as the headache period lasted, and recorded pain and time variables. The severity of PPH was negatively correlated to the time till the headache started or worsened upon rising (T1) and the time from the headache started to increase till it reached its maximum (T2), but was not significantly correlated to the time to restitution upon lying down (T3). The results are in good accordance with the leakage theory. T1 varied from immediate onset to 265 min (median 20 sec). T2 (median 30 sec, range 0-60 min) and T3 (median 20 sec, range 0-15 min) varied considerably as well. During the course of PPH, 45% of the patients occasionally reported non-postural headache or no headache when the Rising Manoeuvre was performed. It is suggested that PPH should be diagnosed in any patient who experiences postural headache at least once within 4 days of lumbar puncture. PMID- 9399010 TI - Ineffectiveness of neurokinin-1 antagonist in acute migraine: a crossover study. AB - Lanepitant is a high-affinity, selective neurokinin-1 receptor (NK-1) and is effective in the dural inflammation model of acute migraine. Lanepitant 30, 80, and 240 mg given orally was evaluated in a double-blind, placebo-controlled crossover study to determine its effect in reducing migraine pain and severity of associated symptoms. Outpatients treated four migraine headaches of moderate or severe pain intensity with study drug according to a randomization schedule. They recorded their pain intensity and severity of migraine-associated symptoms at 30, 60, 90, and 120 min. Although 53 patients were randomly allocated to a treatment sequence, only 40 patients completed all treatments. There was no statistically significant difference in improvement in migraine pain at any time for any of the treatments. Additionally, there was no change in severity of migraine-associated symptoms associated with lanepitant therapy. No adverse events could be attributed to lanepitant. Lanepitant was ineffective orally in treating acute migraine in this trial. This may be due to poor bioavailability during a migraine attack. Alternatively, the neurogenic inflammation hypothesis may not apply to migraine. PMID- 9399011 TI - Zolmitriptan. Introduction. PMID- 9399012 TI - Pre-clinical pharmacology of zolmitriptan (Zomig; formerly 311C90), a centrally and peripherally acting 5HT1B/1D agonist for migraine. AB - Zolmitriptan (Zomig; formerly 311C90) is a novel 5-hydroxytryptamine (5HT)1B/1D receptor agonist with proven efficacy in the acute treatment of migraine with or without preceding aura. The drug differs from presently available members of this drug class in that it combines 5HT1B/1D receptor partial agonist activity with robust oral pharmacokinetics and an ability to inhibit trigeminovascular activation centrally as well as peripherally in preclinical studies. Consistent with its selectivity for 5HT1B/1D receptors, zolmitriptan produces constriction of various isolated blood vessels, most notably cranial arteries. In anaesthetized animals, these vascular effects manifest as a selective constriction of cranial arterio-venous anastomoses resulting in a redistribution of carotid arterial blood flow. This effect is produced without significant effects on heart rate, blood pressure or blood flow to the brain, heart or lungs. Zolmitriptan also inhibits trigeminal-evoked increases in cerebral blood flow in anaesthetized cats and blocks trigeminal-evoked plasma protein extravasation in the dura of guinea-pigs. These actions are consistent with a pre-junctional inhibition of neuropeptide release from perivascular afferents of the trigeminal nerve, as confirmed by independent studies showing that zolmitriptan blocks elevations of calcitonin-gene-related peptide in jugular venous blood during electrical stimulation of the trigeminal ganglion. In all of these effects, zolmitriptan is three to four times more potent than sumatriptan, but produces the same maximum response. Zolmitriptan crosses the intact blood-brain barrier to inhibit trigeminovascular activation in the brainstem. This was shown initially by the ability of the drug to block a brainstem reflex provoking vasoactive intestinal peptide release from the VIIth cranial (facial) nerve during trigeminal stimulation. Subsequent ex vivo autoradiography confirmed that intravenously injected [3H]zolmitriptan labels a discrete population of cells in the trigeminal nucleus caudalis (TNC) and nucleus tractus solitarius. Direct evidence for a central neuromodulatory effect of zolmitriptan was provided by electrophysiological experiments which clearly demonstrated that the drug inhibits the excitability of cells in the TNC after systemic administration. This novel pre-clinical profile not only distinguishes zolmitriptan from sumatriptan, but raises intriguing questions about the clinical relevance of a dual action. Studies to date show that zolmitriptan indeed modulates cranial sensory processing in humans, yet central side-effects are no different from sumatriptan. This property may account for the remarkable consistency in clinical efficacy observed in clinical trials. PMID- 9399013 TI - The clinical pharmacokinetics of zolmitriptan. AB - Zolmitriptan (Zomig, formerly 311C90) is a novel, oral, acute treatment for migraine. In healthy volunteers it is rapidly and extensively absorbed and has favorable oral bioavailability (approximately 40%) which is not affected by concomitant food intake. On average, 75% of its eventual Cmax is achieved within 1 h of dosing. Plasma concentrations are sustained for 4 to 6 h after dosing with single or multiple peaks in the plasma concentration-time profile, reflecting continued absorption down the gastrointestinal tract. The pharmacokinetics of zolmitriptan indicate dose proportionality over the dose range of 2.5 to 50 mg and there are no significant changes on multiple dosing. Zolmitriptan is cleared by metabolism followed by urinary excretion of the metabolites. There are three major metabolites, one of which, the N-desmethyl metabolite, is active as a 5HT1D agonist and has mean plasma concentrations approximately two thirds those of the parent compound. The other two metabolites, the N-oxide and indoleacetic acid, are inactive. The elimination half lives of zolmitriptan and its metabolites are similar, approximately 3 h. Zolmitriptan and its active metabolite are minimally protein bound in the plasma (approximately 25%). In migraine patients, plasma concentrations of zolmitriptan and its metabolites are lower during a migraine attack than outside an attack. In summary, the pharmacokinetics of zolmitriptan are simple, predictable and appropriate to an acute oral treatment for migraine. PMID- 9399014 TI - Potential drug interactions with the novel antimigraine compound zolmitriptan (Zomig, 311C90). AB - Seven randomized studies in healthy volunteers have investigated interactions between zolmitriptan (Zomig, formerly 311C90), a 5HT1B/1D agonist for acute migraine therapy, and selected drugs with which there was a possibility of interaction or a likelihood of concurrent use. Co-administration of oral dihydroergotamine, ergotamine, pizotifen, fluoxetine, paracetamol (acetaminophen)/metoclopramide or selegiline had no clinically significant effects on the pharmacokinetics of zolmitriptan or its metabolites, although small changes were observed in some cases. Co-administration of propranolol resulted in a 56% increase in the area under the plasma concentration-time curve (AUC) of zolmitriptan and a 11% decrease in the AUC of the active metabolite 183C91. However, these pharmacokinetic changes are unlikely to be relevant at lower clinical doses. Moclobemide, a monoamine oxidase A (MAO-A) inhibitor, decreased the clearance of zolmitriptan and, in particular, 183C91. This suggests that MAO-A is involved in the metabolism of 183C91 and it may be prudent to limit the daily zolmitriptan dose in migraine patients maintained on a MAO-A inhibitor. The clinically insignificant blood pressure increases produced by zolmitriptan, and the tolerability profile of this agent, were unaffected by any of the concomitant medications. Clinically significant interactions between zolmitriptan and commonly co-prescribed antimigraine therapies are unlikely. PMID- 9399015 TI - Zolmitriptan (Zomig, 311C90), a novel dual central and peripheral 5HT1B/1D agonist: an overview of efficacy. AB - The efficacy of zolmitriptan (Zomig, 311C90), a 5-hydroxytryptamine (5HT)1B/1D receptor agonist, in the acute oral treatment of migraine was evaluated in an extensive clinical trial program. Four randomized, placebo-controlled studies (total 2480 patients) were performed; data from two of these trials established that a 2.5 mg dose was on the shoulder of the dose-response curve (2-h headache response rate 64%), showing similar efficacy to the 5 mg dose (67%). In this program, the efficacy of zolmitriptan was not influenced by the pretreatment headache duration; the presence of aura preceding the headache, migraine associated with menses or migraine upon awakening; or by concomitant use of oral contraceptives or antidepressants. In addition, zolmitriptan 5 mg proved consistently effective in the treatment of multiple migraine attacks for up to 1 year. Zolmitriptan reduced the incidence of nausea, photophobia and phonophobia, reduced impairment of normal activity and demonstrated positive effects on patients' quality of life. Thus, zolmitriptan is a highly effective acute oral antimigraine therapy, with 2.5 mg providing the optimal balance between efficacy and tolerability. PMID- 9399016 TI - Tolerability profile of zolmitriptan (Zomig; 311C90), a novel dual central and peripherally acting 5HT1B/1D agonist. International clinical experience based on > 3000 subjects treated with zolmitriptan. AB - Zolmitriptan (Zomig, formerly 311C90) at doses of 0.5-50 mg was administered to 316 unique volunteers in clinical pharmacology studies and 2,750 unique patients in eight clinical studies of acute migraine treatment. Overall, subjects received almost 50,000 doses; 97% of exposures were at doses > or = 2.5 mg. In the clinical pharmacology studies, the overall incidence of subject exposures experiencing at least one adverse event was 52% with zolmitriptan 2.5 mg (28% with placebo). In placebo-controlled studies, the overall incidence of patients with at least one adverse event was dose-dependent for zolmitriptan over the 1-15 mg dose range, e.g. 42% and 46% with 1 and 2.5 mg, respectively and 58% with 5 mg (29% with placebo). Only four serious adverse events attributable to zolmitriptan were reported. In a long-term study, during which 2,058 outpatients treated a total of 31,579 migraine attacks with either one or two zolmitriptan 5 mg doses over a period of up to 1 year, the number of attacks associated with at least one adverse event was similar after one (26%) and two (24%) doses. The majority (59%) of the adverse events reported in this study (59%) occurred within 2 h of dosing, were predominantly mild (59%) or moderate (35%) in intensity, of < or = 4 h duration (58%), required no further action (94%). In placebo-controlled studies, the percentage of patients who reported severe adverse events was similar with zolmitriptan 2.5 mg (4%) and placebo (5%). The most frequently reported adverse events with zolmitriptan in the placebo-controlled clinical studies were asthenia, heaviness (other than chest or neck), dry mouth, nausea, dizziness, somnolence, paresthesia and warm sensations. The type and severity of the adverse events was not influenced by gender (although the frequency of reported adverse events was higher in females, as was the case in the placebo group), age, presence of aura prior to the attack, association of migraine with menstruation, concurrent medication, or by the addition of a second zolmitriptan dose. Zolmitriptan showed a similar tolerability profile in the long-term study, in which a low withdrawal rate due to adverse events of 8% was observed. Zolmitriptan was not associated with an increased frequency of central nervous system-related adverse events in a comparative study of sumatriptan, despite pre clinical and neurophysiological evidence of a dual peripheral and central action of zolmitriptan. Moreover, zolmitriptan doses of 5-20 mg produced no statistically significant effects on objective assessments of psychometric function. Zolmitriptan had no clinically significant effects on blood pressure (even in patients with controlled mild to moderate hypertension or impaired renal function), ECGs (e.g. there was no evidence of ischemic events) or clinical chemistry, hematological or urinalysis measurements. In summary, zolmitriptan is well tolerated, particularly at the recommended dose of 2.5 mg. Zolmitriptan has a well-defined dose-response with 2.5 mg proving highly effective and optimizing the benefit/risk ratio of treatment. Thus, zolmitriptan is well suited as an acute oral treatment for migraine in the outpatient setting. PMID- 9399017 TI - Clinical applications of zolmitriptan (Zomig, 311C90). AB - Zolmitriptan (Zomig, formerly 311C90) is a novel, oral antimigraine drug that is consistently effective and well tolerated in the acute treatment of migraine headache and its associated symptoms. The purpose of this article is to review data available from pharmacological and clinical trials with zolmitriptan and to summarize the clinically relevant features that distinguish this agent. We will review the attributes desirable in a migraine drug and use this as a template for assessing zolmitriptan. Zolmitriptan provides a new oral treatment option for physicians which should help improve patient outcomes. PMID- 9399018 TI - Diabetes-induced alterations in platelet metabolism. AB - OBJECTIVES: This review summarizes the recent findings on some aspects of platelet metabolism that appear to be affected as a consequence of diabetes mellitus. The metabolites include glutathione, L-Arginine/nitric oxide, as well as the ATP-dependent exchange of Na+/K+ and Ca2+. CONCLUSIONS: Several aspects of platelet metabolism are altered in diabetics. These metabolic events give rise to a platelet that has less antioxidants, and higher levels of peroxides. The direct consequence of this is the overproduction platelet agonists. In addition, there is evidence for altered Ca2+ and Na+ transport across the plasma membrane. Recent evidence indicates that plasma ATPases in diabetic platelets are not damaged instead their activities are likely to be modulated by oxidized LDL. Finally, platelet inhibitory mechanisms regulated by NO appear to be perturbed in the diabetes disease-state. The combined production of NO and superoxide by NOS isoforms in the platelet could be a major contributory factor to platelet pathogenesis in diabetes mellitus. PMID- 9399019 TI - Reverse cholesterol transport--a review of the process and its clinical implications. AB - OBJECTIVES: This review article will summarize the current knowledge surrounding the reverse cholesterol transport system; the process, the effect of mutations in genes coding for proteins which function in the system, and the possible clinical implications of these alterations. RESULTS: High-density lipoprotein-cholesterol (HDL-C) concentration is a marker for the reverse cholesterol transport (RCT) system, whereby cholesterol is returned from peripheral cells to the liver for reuse or excretion in the bile. Increased HDL-C concentrations are generally accepted to be protective against the future development of atherosclerosis and coronary artery disease (CAD), but recent evidence has indicated that the underlying cause of the increased HDL-C may affect whether it is protective or detrimental. The major steps in the RCT pathway are the efflux of free cholesterol from cells and binding by pre-beta HDL, esterification of HDL-bound cholesterol by lecithin cholesterol acyl transferase (LCAT), cholesteryl ester transfer protein (CETP) mediated exchange of cholesteryl ester and triglycerides between HDL and apo B-containing particles, and hepatic lipase (HL) mediated uptake of cholesterol and triglycerides by the liver. Mutations in proteins active in the RCT pathway can shed light on the functions and control of the various steps in the system. LCAT deficiency, leading to greatly reduced HDL and fish eye disease, is not usually associated with increased risk of CAD. Several new mutations in LCAT have recently been reported, however, which do result in CAD. Mutations leading to reduced CETP activity result in less CE being directed into apo-B containing particles and more remaining in the HDL. This has been associated with increased HDL-C concentrations. The generally accepted hypothesis that reduced CETP activity leads to reduced CAD risk has been challenged by a number of recent publications, and has become an area of active investigation. Mutations leading to reduced HL activity are rare occurrences. To date, all have been associated with increased HDL-C concentrations and CAD. CONCLUSION: The development of techniques to identify and characterize the functional significance of mutations in proteins involved in RCT will aid in the understanding of the mechanisms and control of this pathway. PMID- 9399020 TI - Effect of repeated freeze-thaw cycles on maternal serum biological markers for the detection of fetal trisomies. AB - OBJECTIVES: We studied the stability of maternal blood markers for screening for Down syndrome (alpha-fetoprotein, unconjugated estriol, intact human chorionic gonadotropin (hCG) and free beta-human chorionic gonadotropin) upon repeated freeze-thaw cycles. DESIGN AND METHODS: Forty-three samples collected from second trimester normal pregnancies were submitted to five freeze-thaw cycles. After each cycle, the markers were measured using kits and instruments from Wallac Canada (AutoDelfia). Results were compared by repeated measures analysis of variance and by analysis of linear trend (after mathematical transformation of the results in order to decrease between-sample variation) as a function of the number of freeze-thaw cycles. RESULTS: No significant differences were observed by ANOVA (p > 0.1) for any marker. Intact hCG showed a statistically significant linear downward trend (slope = -0.0063, p = 0.009) while free beta-hCG increased (slope = 0.011, p = 0.004). After five freeze-thaw cycles, a mean decrease of 3.2% is predicted for intact hCG while free beta-hCG would increase by 5.5% on average. CONCLUSION: We conclude that the studied markers do not show clinically significant changes under the evaluated conditions. The observed changes of intact hCG and free beta-hCG would have a limited impact on the screening performance. PMID- 9399021 TI - LDL receptor activity in human leukocyte subtypes: regulation by insulin. AB - OBJECTIVES: LDL receptors of leukocytes play a key role in lipoprotein uptake, immunoregulation and the pathogenesis of atherosclerosis. Numerous studies with different methods of low reliability yielded conflicting results of its regulation in leukocyte subtypes. DESIGN AND METHODS: LDL receptors of human leukocytes were measured with use of the monoclonal antibody C-7. Specific C-7 binding was detected by FACS analysis using phycoerythrin-anti-mouse-IgG. Parallel incubations with FITC-labelled anti-LEU 4 (CD 3), anti-LEU 12 (CD 19) and anti-MY 4 (CD 14) antibodies were used to distinguish C-7 binding of specific cell types (T-, B-lymphocytes and monocytes). RESULTS: In contrast to monocytes, T and B-lymphocytes freshly isolated from healthy blood donors had no detectable binding capacity for C-7. After 24 and 48 h incubation of cells in a lipid-free medium, lymphocytes acquired some C-7 binding, albeit still much less than monocytes. Incubation with insulin for 24 h in a concentration of 0.5 microgram/mL led to an increase in C-7 binding for monocytes (up to 180%). Saturation experiments with the ligand suggests an increase in the number of receptors. In contrast the same insulin concentration inhibited C-7 binding of B- and T-lymphocytes by 35%. CONCLUSIONS: FACS analysis using monoclonal antibodies seems to be a feasible method for the investigation of lipid metabolism in leukocytes. The LDL receptor expression and its regulation by insulin differs in circulating monocytes and lymphocytes. PMID- 9399022 TI - Radioreceptor assay for sirolimus in patients with decreased platelet counts. AB - OBJECTIVE: Sirolimus (RAPA) is a new immunosuppressive drug currently in Phase III clinical trials in combination with cyclosporine A (CsA). The toxicity profiles for CsA and RAPA are only partially overlapping, with RAPA toxicity consisting primarily of hyperlipidemia and myelodepression but without the nephrotoxicity, neurotoxicity, and hepatotoxicity, which are seen with CsA. Patients in the clinical trial are being monitored using HPLC or LC/MS/MS assays; there is no immunoassay for RAPA reported to date. We have previously reported a radioreceptor assay (RRA) for RAPA, which has an excellent correlation with the HPLC assay (r = 0.997). The RRA has several advantages including excellent precision, sensitivity, rapid turnaround time, and a one-step extraction procedure. We report the evaluation of blood samples from patients who were exhibiting RAPA toxicity and comparison of the RRA results with the HPLC results. METHODS: EDTA whole blood specimens (n = 42) were obtained from six renal transplant recipients taking RAPA and CsA and exhibiting decreased platelet counts. Thirty-two samples from patients without decreased platelet counts were also received. The samples were analyzed with the RRA and the results were compared to those obtained with the HPLC assay. RESULTS: By HPLC, the results ranged from 3.2-72.6 micrograms/L RAPA with 43% of the results > or = 30 micrograms/L. With the RRA, the range was 7.7-83.0 micrograms/L RAPA equivalents, with 60% of the results > or = 30 micrograms/L. The RRA results are distinctly higher than the HPLC results all along the range. The correlation between the two assays was 0.861, with a slope of 0.966 and a Y-intercept of 11.1. CONCLUSION: Since the RRA is consistently higher than HPLC concentration in patients with decreased platelet counts, but correlates well in patients with no signs of toxicity, the RRA may be useful for monitoring patients for toxicity, by giving a better indication of increasing degree of immunosuppression than the HPLC assay. PMID- 9399023 TI - Intestinal alkaline phosphatase isoforms in rabbit tissues differ in glycosylation patterns. AB - OBJECTIVES: In patients with advanced liver cirrhosis or chronic nephritis, an intestinal alkaline phosphatase (IAP)-like enzyme is ectopically expressed in the liver or kidney. In this study, we used rabbit organs as a human pathological model, because the rabbit liver or kidney expresses an IAP-like enzyme as the predominant isozyme, unlike humans. METHODS: IAP and the IAP-like enzyme were purified from rabbit intestine and kidney, respectively, by immunoaffinity chromatography using a monoclonal antihuman IAP antibody. Some properties of the IAP and IAP-like enzyme expressed in rabbit organs are compared. RESULTS: Some of their catalytic and physicochemical properties differed. In particular, the net charge, molecular mass, and hydrophobicity of IAP from rabbit intestine was slightly different from the IAP-like enzyme from rabbit kidney. There was a difference in the sugar chain structure between the two enzymes according to the results of lectin affinity chromatography, and part of the peptide maps differed slightly. However, there was no difference in the peptide maps after treatment with endo-N-acetylglucosaminidase F. CONCLUSIONS: The primary structures of IAP and the IAP-like enzyme are basically similar, except for the glycosylation process of the respective AP isozymes. PMID- 9399025 TI - Plasma diamine oxidase activities in renal dialysis patients, a human with spontaneous copper deficiency and marginally copper deficient rats. AB - OBJECTIVES: Intestine and kidney are generally the most concentrated sources of the copper metalloenzyme diamine oxidase (DAO). Clinically, plasma DAO activities are used to diagnose disruptions in intestinal integrity. This study determined whether DAO activities were also affected by kidney injury or copper nutritional status. DESIGN AND METHODS: Plasma DAO activities were measured in renal dialysis patients without diagnosed intestinal disease (n = 75), controls (n = 23), an adult with spontaneous copper deficiency before and after copper repletions, and in rats fed either adequate or marginal copper diets (8 or 2 mg copper/kg diet) for 7 months. RESULTS: This study found high DAO activities in renal dialysis patients and low activities during spontaneous copper deficiency. Low activities were also seen for marginally copper deficient rats. CONCLUSIONS: Tissue injury induced elevation of DAO activities is not limited to intestinal injury, and low DAO values may be useful for assessing copper nutritional status. PMID- 9399024 TI - Plasma total antioxidant capacity in an adult Hong Kong Chinese population. AB - OBJECTIVES: To examine plasma total antioxidant capacity (TAOC) in a Chinese population with a lower incidence of cardiovascular disease compared with Caucasian populations, in relation to dietary intake, age, sex, and the presence of cardiovascular diseases. DESIGN AND METHODS: As part of a randomized territory wide survey stratified by sex and 10-year age groups; 728 subjects (367 men, 361 women) were recruited. Dietary intake assessment was by a food frequency questionnaire; plasma TAOC was estimated by the ABTS method. RESULTS: The TAOC values were normally distributed, the mean +/- SD being 1.78 +/- 0.18 mmol/L. The mean value was higher in men compared with women, inspite of a lower dietary intake of vitamins A and C per 1000 kcal in the former. Subjects who consumed water spinach twice or more a week had higher mean levels. No difference in mean levels was observed between those with and without hypertension or cardiovascular disease. CONCLUSION: Measurement of plasma TAOC as a risk factor in epidemiological studies of cardiovascular diseases may have limited use, since TAOC include substances associated with a protective effect as well as increased risk. PMID- 9399026 TI - Analysis of sertraline and desmethylsertraline in human plasma and red blood cells. PMID- 9399027 TI - Serum Mn-superoxide dismutase in acute myocardial infarction. PMID- 9399028 TI - Hereditary kidney diseases. Introduction. PMID- 9399029 TI - Pathogenesis of autosomal dominant polycystic kidney disease: recent developments. PMID- 9399030 TI - Autosomal recessive polycystic kidney disease. PMID- 9399031 TI - Prevalence of hypertension according to phenotype and gender in autosomal dominant polycystic kidney disease. PMID- 9399032 TI - Role of renin-angiotensin-aldosterone system and of sympathetic activity in arterial hypertension associated with autosomal dominant polycystic kidney disease. PMID- 9399033 TI - Hypertension in polycystic kidney disease type 1 and 2 and its effect on the age of onset of end-stage renal disease. PMID- 9399034 TI - Sodium-lithium countertransport in autosomal polycystic kidney disease. PMID- 9399035 TI - Inflammatory cytokine profile in autosomal dominant polycystic kidney disease. PMID- 9399036 TI - Extracellular matrix abnormality may be responsible for cyst development. PMID- 9399037 TI - Mutations and intragenic polymorphisms in the diagnosis of autosomal dominant polycystic kidney disease type 1. PMID- 9399038 TI - Expression of protein fragments from the human PKD1 gene and production of rabbit polyclonal antibodies to the recombinant proteins. PMID- 9399039 TI - Molecular genetic investigations in autosomal dominant polycystic kidney disease. Gene Mutation detection, linkage analysis, and preliminary ACE gene I/D polymorphism association studies: an update. PMID- 9399040 TI - Oral-facial-digital syndrome type 1 coexisting with polycystic kidney disease. PMID- 9399041 TI - Nephronophthisis-medullary cystic kidney disease complex: a report on 24 patients from 5 families with Italian ancestry. PMID- 9399042 TI - Renal manifestations of tuberous sclerosis complex. PMID- 9399043 TI - The TSC2/PKD1 contiguous gene syndrome. PMID- 9399044 TI - A case of Pringle-Bourneville tuberous sclerosis with renal angiomyolipomas: clinical and radiological aspects. PMID- 9399045 TI - Tuberous sclerosis and nephrocalcinosis. PMID- 9399046 TI - A tuberous sclerosis patient with a large TSC2 and PKD1 gene deletion shows extrarenal signs of autosomal dominant polycystic kidney disease. PMID- 9399047 TI - Tuberous sclerosis complex and early-onset autosomal dominant polycystic kidney disease as a 'contiguous gene' syndrome: report of a case. PMID- 9399048 TI - Pulmonary lymphangioleiomyomatosis and tuberous sclerosis complex. PMID- 9399049 TI - The kidney and von Hippel-Lindau disease: impact of molecular genetic analysis of the VHL gene for clinical management. PMID- 9399050 TI - Von Hippel-Lindau (VHL) gene analysis in Italian families with VHL disease. PMID- 9399051 TI - Planning kidney surgery in von Hippel-Lindau disease. PMID- 9399052 TI - Alport syndrome: clinical and genetic correlation in a type-IV collagen disease. PMID- 9399053 TI - Clinical and molecular diagnosis in inherited kidney diseases: three examples. PMID- 9399054 TI - Expression of alpha (IV) chains in Alport's syndrome and its correlation with ultrastructural and genetic data. PMID- 9399055 TI - Molecular diagnosis of Alport syndrome: the experience in Siena. PMID- 9399056 TI - X-linked Alport syndrome with normal distribution of collagen IV alpha chains in epidermal basement membrane. PMID- 9399057 TI - Kidney transplantation in Alport's syndrome. PMID- 9399058 TI - Primary hyperoxaluria. PMID- 9399059 TI - Renal pathology in hyperoxaluria. PMID- 9399060 TI - Clinical aspects of cystinuria. PMID- 9399061 TI - Cystinuria: recent advances in pathophysiology and genetics. PMID- 9399062 TI - Kidney involvement in Anderson-Fabry disease. PMID- 9399063 TI - Anderson-Fabry disease. Three families detected in 2 years: unusual occurrence or good interdisciplinary collaboration? PMID- 9399064 TI - Genetic approach to the study of cellular ion transport anomalies in idiopathic calcium nephrolithiasis. PMID- 9399066 TI - Familial hemolytic uremic syndrome: simulation linkage analysis. PMID- 9399065 TI - Constitutive nitric oxide synthase gene expression in Bartter's and Gitelman's syndrome and its relationship to their vascular hyporesponsiveness. PMID- 9399067 TI - Abnormal blood glucose and insulin response during oral glucose tolerance test in familial renal glycosuria. PMID- 9399068 TI - Renal transplantation in patients with hereditary kidney disease: our experience. PMID- 9399069 TI - An information center for rare diseases: a tool for epidemiologic and clinical studies in rare diseases. PMID- 9399070 TI - Transcriptional regulation of vertebrate Hox genes during embryogenesis. AB - The identification in transgenic mice of Hox gene DNA regulatory elements that can recapitulate certain aspects of the endogenous gene's expression pattern has proceeded with great success. However, perfect reproduction of the correct expression pattern is uncommon, even when large genomic fragments spanning neighboring genes are analyzed, suggesting that important regulatory regions may be located at large distances from the genes they control or that their specific context may be important. Currently, four groups of transcriptional regulators have been identified that have been shown to directly regulate Hox gene expression in the vertebrate embryo: retinoic acid receptors, Krox20, members of the Pbx/exd family, and the Hox genes themselves. PMID- 9399071 TI - Histone H1 and chromatin higher-order structure. AB - The linker histone H1, or its variants such as H5, have long thought to be involved in promoting the organization of chromatin into a higher-order structure, the 30 nm filament. However, the location of H1 in the filament, its role in filament formation and the structure of the 30 nm filament itself have all been controversial. This article reviews recent results that address these questions. PMID- 9399072 TI - Application of atomic force microscopy to visualization of DNA, chromatin, and chromosomes. AB - The scanning force microscope (SFM, also called the atomic force microscope, AFM) provides a new and powerful method for visualization and manipulation of biological samples. Its high precision and sensitivity allow the investigator to interrogate samples at very high spatial resolution and simultaneously accumulate a variety of data types, including topography, viscoelasticity, chemical properties, and local friction. We provide here a brief review of the literature describing the current state of the art in the application of SFM to the study of DNA, chromatin and chromosomes, and some examples from this laboratory. Suggestions for future directions of this technology are also presented. PMID- 9399073 TI - The regulation of clotting factors. AB - Blood clotting involves a multitude of proteins that act in concert in response to vascular injury to produce the procoagulant enzyme alpha-thrombin, which in turn is responsible for the generation of the fibrin plug. However, while generation of the fibrin plug is required for the arrest of excessive bleeding, unregulated clotting will result in the occlusion of the blood vessels and thrombosis. Thus, the regulation of the delicate balance between the procoagulant and anticoagulant mechanisms is of extreme importance for survival. While the majority of proteins involved in blood coagulation circulate as inactive zymogens that require proteolytic activation in order to function, approximately 1% of the circulating factor VII molecules are active. Factor VIIa, possess a serine protease active site, has poor catalytic activity, and is not inhibited by the circulating stoichiometric protease inhibitors. Following injury to the vasculature and subsequent exposure of the integral membrane glycoprotein, tissue factor (TF), the circulating factor VIIa molecules can bind to the exposed TF forming the extrinsic tenase complex (TF/factor VIIa) and initiate the blood coagulation process. Formation of the TF/factor VIIa complex increases the catalytic efficiency of the enzyme by four orders of magnitude when compared with factor VIIa alone. This cell-associated enzymatic complex initiates a series of enzymatic reactions, leading to the generation of alpha-thrombin and ultimately to the formation of the fibrin plug. The procoagulant enzymatic complexes (i.e., prothrombinase, intrinsic tenase, and extrinsic tenase) are similar in structure and composed of an enzyme, a cofactor, and the substrate associated on a cell surface in the presence of divalent metal ions. While the activity of the extrinsic tenase complex is limited by the availability (exposure) of its cell associated cofactor (TF) it is remarkable that the activities of both the prothrombinase complex (factor Va/factor Xa) as well as the intrinsic tenase complex (factor VIIIa/factor IXa) are limited by the presence of the two soluble, nonenzymatic cofactors, factor Va and factor VIIIa. Factor Va and factor VIIIa, which are very similar in structure and function, are required for prothrombinase and intrinsic tenase activities, respectively, because both cofactors express a dual function in their respective complexes, acting as an enzyme receptor and catalytic effector on the cell surface. The cofactors derive from inactive plasma precursors by regulatory proteolytic events that involve alpha-thrombin. In general, bleeding tendencies are usually associated with defects in the activation of one of the zymogens or the cofactors of the procoagulant complexes. However, the activity of all of the complexes is also limited by the availability of an adequate membrane surface provided by endothelial cells, platelets, and monocytes. The cell surface provides a site for the recruitment of the appropriate proteins and allows for fast and efficient clot formation. In the absence of an appropriate membrane surface, the procoagulant complexes have limited catalytic efficiency. Thus, timely exposure of the adequate membrane surface is an additional step in the regulation of alpha-thrombin formation. alpha-Thrombin participates in its own down-regulation by binding to the endothelial cell receptor thrombomodulin, initiating the protein C pathway, which in turn leads to the formation of activated protein C (APC). APC is required for efficient neutralization of factor Va cofactor activity, which results in the inactivation of the prothrombin-activating complex. This inactivation can only occur in the presence of the appropriate membrane surface. Thus, while following alpha-thrombin activation, factor VIIIa is rapidly and spontaneously inactivated by dissociation of the A2 domain from the rest of the cofactor, APC is required for down-regulation of alpha-thrombin formation by prothrombinase. (ABSTRACT PMID- 9399074 TI - Vasculitis in children: a diagnostic challenge. PMID- 9399075 TI - Paternal effects in Drosophila: implications for mechanisms of early development. AB - The study of paternal effects on development provides a means to identify sperm supplied products required for fertilization and the initiation of embryogenesis. This review describes paternal effects on animal development and discusses their implications for the role of the sperm in egg activation, centrosome activity, and biparental inheritance in different animal species. Paternal effects observed in Caenorhabditis elegans and in mammals are briefly reviewed. Emphasis is placed on paternal effects in Drosophila melanogaster. Genetic and cytologic evidence for paternal imprinting on chromosome behavior and gene expression in Drosophila are summarized. These effects are compared to chromosome imprinting that leads to paternal chromosome loss in sciarid and coccid insects and mammalian gametic imprinting that results in differential expression of paternal and maternal loci. The phenotypes caused by several early-acting maternal effect mutations identify specific maternal factors that affect the behavior of paternal components during fertilization and the early embryonic mitotic divisions. In addition, maternal effect defects suggest that two types of regulatory mechanisms coordinate parental components and synchronize their progression through mitosis. Some activities are coordinated by independent responses of parental components to shared regulatory factors, while others require communication between paternal and maternal components. Analyses of the paternal effects mutations sneaky, K81, paternal loss, and Horka have identified paternal products that play a role in mediating the initial response of the sperm to the egg cytoplasm, participation of the male pronucleus in the first mitosis, and stable inheritance of the paternal chromosomes in the early embryo. PMID- 9399076 TI - Drosophila myogenesis and insights into the role of nautilus. AB - Several aspects of muscle development appear to be conserved between Drosophila and vertebrate organisms. Among these is the conservation of genes that are critical to the myogenic process, including transcription factors such as nautilus. From a simplistic point of view, Drosophila therefore seems to be a useful organism for the identification of molecules that are essential for myogenesis in both Drosophila and in other species. nautilus, the focal point of this review, appears to be involved in the specification and/or differentiation of a specific subset of muscle founder cells. As with several of its vertebrate and invertebrate counterparts, it is capable of inducing a myogenic program of differentiation reminiscent of that of somatic muscle precursors when expressed in other cell types. We therefore favor the model that nautilus implements the specific differentiation program of these founder cells, rather than their specification. Further analyses are necessary to establish the validity of this working hypothesis. Studies have revealed a critical role for Pax-3 in specifying a particular subset of myogenic cells, the progenitors of the limb muscles. These myogenic cells migrate from the somite into the periphery of the organism, where they differentiate. These myoblasts do not express MyoD or myf5 until they have arrived at their destination and begin the morphologic process of myogenesis (Bober et al., 1994; Goulding et al., 1994; Williams and Ordahl, 1994). They then begin to express these genes, possibly to put the myogenic plan into action. Thus, as with nautilus, MyoD and myf5 may be necessary for the manifestation of a muscle-specific commitment that has already occurred. By comparison with vertebrates, it was anticipated that the single Drosophila gene would serve the purpose of all four vertebrate genes. However, its restricted pattern of expression and apparent loss-of-function phenotype are inconsistent with this expectation. It remains to be determined whether nautilus functions in a manner similar to just one of the vertebrate genes. Since the myf5- and MyoD-expressing myoblasts are proliferative, the loss of one cell type appears to be compensated by proliferation of the remaining cell type. This apparent plasticity may obscure differences in mutant phenotype resulting from the loss of particular cells that express each of these genes. In Drosophila, by comparison, nautilus-expressing cells committed to the myogenic program undergo few, if any, additional cell divisions, and thus no other cells are available to compensate for the loss of nautilus. Therefore, the apparent differences between the Drosophila nautilus gene and its vertebrate counterparts may reflect, at least in part, differences in the developmental systems rather than differences in the function of the genes themselves. PMID- 9399077 TI - Hydrozoa metamorphosis and pattern formation. PMID- 9399078 TI - Primate embryonic stem cells. AB - Primate embryonic stem (ES) cells are derived from preimplantation embryos, have a normal karyotype, and are capable of indefinite, undifferentiated proliferation. Even after culture for more than a year, primate ES cells maintain the potential to differentiate to trophoblast and derivatives of embryonic endoderm, mesoderm, and ectoderm. In this review, we compare the characteristics of ES cell lines from two primate species, the rhesus monkey (Macaca mulatta) and the common marmoset (Callithrix jacchus), with the characteristics of mouse ES cells and human embryonal carcinoma cells. We also discuss the implications of using primate ES cells to understand early human development and discuss the practical and ethical implications for the understanding and treatment of human disease. PMID- 9399079 TI - Sex determination in plants. AB - The majority of flowering plants produce flowers that are "perfect." These flowers are both staminate (with stamens) and pistillate (with one or more carpels). In a small number of species, there is spatial separation of the sexual organs either as monoecy, where the male and female organs are carried on separate flowers on the same plant, or dioecy, where male and female flowers are carried on separate male (staminate) or female (pistillate) individuals. Sex determination systems in plants, leading to unisexuality as monoecy or dioecy, have evolved independently many times. In dioecious plant species, the point of divergence from the hermaphrodite pattern shows wide variation between species, implying that the genetic bases are very different. This review considers monoecious and dioecious flowering plants and focuses on the underlying genetic and molecular mechanisms. We propose that dioecy arises either from monoecy as an environmentally unstable system controlled by plant growth substances or from hermaphroditism where the underlying mechanisms are highly stable and control does not involve plant growth substances. PMID- 9399080 TI - Somitogenesis. AB - We are still far from understanding "somitogenesis" as a whole, but there is an emerging picture of the tissue interactions and molecular mechanisms that underlie and govern various aspects of this essential multistep patterning process in vertebrates. The ability to form segmental units appears to be a property specific to the paraxial mesoderm (as opposed to lateral or limb mesoderm), and this ability is probably acquired during early development, when paraxial mesoderm is specified and emerges from the primitive streak. Signaling molecules expressed in the primitive streak and tail bud are prime candidates involved in specifying paraxial (as well as other mesodermal) fates. Increasing levels of signaling molecules may be required in posterior regions of the embryo, and combinatorial signals may be essential to specify the paraxial mesoderm along the entire anterior-posterior axis. However, most of the pivotal signals, and the ways in which they are integrated and interact, remain enigmatic. Once the paraxial mesoderm is formed, segmentation proceeds largely without the requirement for continuous interactions with surrounding tissues. Somitomeres represent a morphologic pattern in the mesenchymal presomitic mesoderm, but their significance for somite formation is unclear. Molecular patterns are established in the presomitic mesoderm and probably are of functional significance. Cell interactions within the paraxial mesoderm appear to be involved in forming segment borders and ensuring their maintenance during subsequent differentiation of somites. These interactions are, at least in part, mediated by components of the conserved Notch signaling pathway, which may have multiple functions during somitogenesis. Epithelial somites are clearly a result of segmentation, but epithelialization is not the mechanism to form segments, supporting the idea that the basic mechanisms that govern segmentation in the mesoderm of vertebrates are very similar in different species despite divergent types of resulting segments (i.e., epithelial somites versus rotated myotomes). Concomitantly with segmentation, segment polarity and positional specification are established. How these processes are linked to, and depend on, each other is unknown, as is how they are regulated and how segmentation is coordinated on both sides of the neural tube. In contrast to early patterning in the presomitic mesoderm, patterning of the mature somites during their subsequent differentiation is the result of extensive tissue interactions. Virtually all tissues in close proximity to somites provide signals that are involved in induction or inhibition of particular differentiation pathways, but how these pathways are initiated is less clear. Some of the molecules mediating inductive signals and tissue interactions are known, and a growing number of candidate genes are potentially involved in regulating various steps of somitogenesis. The roles of these genes have yet to be analyzed. In addition, the molecular genetic analysis of mutations affecting somitogenesis, which were collected in the mouse and more recently in the zebrafish (Driever et al., 1996; Haffter et al., 1996; van Eeden et al., 1996), promises to add important new insights into this process. Much remains to be done, but the tools are at hand to provide further understanding of the molecular mechanisms underlying somitogenesis. PMID- 9399081 TI - Improving control of patient status in critical care. PMID- 9399082 TI - A clinical description of the IMPROVE Data Library. PMID- 9399083 TI - Building the IMPROVE Data Library. PMID- 9399084 TI - Collecting EEG signals in the IMPROVE Data Library. PMID- 9399085 TI - Using artificial neural networks for classifying ICU patient states. PMID- 9399086 TI - Data fusion of electrophysiological and haemodynamic signals for ventricular rhythm tracking. PMID- 9399087 TI - Signal processing in prolonged EEG recordings during intensive care. PMID- 9399088 TI - Monitoring the autonomic nervous system in the ICU through cardiovascular variability signals. PMID- 9399089 TI - Evaluating time-varying heart-rate variability power spectral density. PMID- 9399090 TI - A portable monitor for fetal heart rate and uterine contraction. PMID- 9399091 TI - Locomotion and steering aspects in automation of colonoscopy. Part one.A literature review. PMID- 9399093 TI - Bowel-sound signal enhancement using adaptive filtering. PMID- 9399092 TI - Relative and intermittent cardiorespiratory coordination. PMID- 9399094 TI - Patenting biotechnology: ethical and philosophical issues. PMID- 9399095 TI - Whitaker Foundation funds genome-related research. PMID- 9399096 TI - Identification of localized anti-host responses in the graft mesenteric lymph node and Peyer's patches after rat small bowel transplantation. AB - This study determined the activation status of recipient and donor lymphocyte populations in the graft mesenteric lymph node (MLN) and Peyer's patches (PP) after allogeneic, heterotopic rat small bowel transplantation without immunosuppression. Untransplanted and isografted animals served as controls. The activation status of lymphocyte subsets was determined by flow cytometric evaluation of lymphoblastoid transformation (forward light scatter; FSc). The proportion of activated lymphocytes in the MLN and PP of allografted animals progressively increased. There was also an early transient activation of MLN lymphocytes in isografted animals which probably resulted from surgery-related inflammation. Activated alpha/beta TCR+ and CD4+ cells were detected in the MLN as early as day 3, whereas there was little activation of CD8+ cells. Interestingly, donor lymphocytes became more activated than recipient lymphocytes. Allografting also led to activation of graft-derived PP alpha/beta TCR+ and CD8+ cells, yet there was no detectable activation of recipient-derived lymphocytes. In summary, this study has identified activated donor lymphocytes in the graft MLN and PP after allogeneic small bowel transplantation. Although rejection predominates without immunosuppression, the presence of an underlying anti-recipient response within the small bowel allograft may contribute to graft damage via the localized release of cytokines and inflammatory mediators. PMID- 9399097 TI - Identification of core B cell epitope in the synthetic peptide inducing cross inhibiting antibodies to a surface protein antigen of Streptococcus mutans. AB - A surface protein antigen (PAc) of Streptococcus mutans, in particular, A-region of the molecule, has been considered as a possible target for the development of an effective anticaries vaccine. This region might be implicated in the induction of dental caries via interaction with salivary components. We have recently specified a unique peptide, TYEAALKQYEADL, as one of the minimum peptides that completely corresponds to the amino acid sequence of a part of the A-region. The unique peptide contains both T and B cell epitopes for the induction of cross reacting antibodies to the PAc. In this study, we synthesized valine or glycine substituted peptide analogs of this peptide and examined core B cell epitopes of this unique peptide by using ELISA inhibition assay. As a result, the core amino acid residues of -Y------Y---- for B cell recognition were found to likely be not only important amino acids stabilizing the structure, but also might be essential for induction of the cross-inhibiting antibodies against PAc. These results will hopefully provide us with useful information for the design of an effective anticaries peptide vaccine. PMID- 9399098 TI - Comparison of different immunoenzymatic methods for the determination of the fine specificity and affinity constants of polyclonal antibodies against pseudopeptide haptens. AB - We evaluated two phosphinopeptides and one phosphonopeptide, which are transition state analogs of a proteolytic reaction, for their ability to generate murine polyclonal antibodies. The specificity of these antisera was determined by indirect and competitive ELISA. Cross-reactivity analysis by these ELISA showed that the antisera recognized selectively haptens containing a phosphate group. The pseudopeptides recognized by the antisera in the indirect ELISA were not the same, however, as those recognized in the competitive ELISA. The differences between these results are probably due to the presentation forms of the hapten, i.e., passively adsorbed in the former ELISA format and soluble in the latter. The affinity of the antibodies was then determined by using two methods based on competitive ELISA, one described by Friguet et al. and the other by Seligman. The dissociation constant (Kd) values calculated by the two methods, for an antiserum and its homologous hapten, are similar. However, only the middle portion of the inhibition scale in Seligman's method gave access to reliable values. Nevertheless, the Seligman representation allowed us to underscore the large range of affinity constants of the polyclonal antibodies. PMID- 9399099 TI - In vitro suppression of the normal mitogenic T lymphocyte response by steady state sickle cell disease sera. AB - This study is part of a long term evaluation of sickle cell disease (SCD) as a paradigm for immunosuppression. Serum was obtained from 43 SCD patients during the steady (healthy) state. Peripheral blood mononuclear cells (PBMC), separated by density gradient were obtained from 8 normal healthy donors. PBMC were utilized in assays directly or as a source for obtaining, total T (CD3) and helper T (CD4) cell populations separated by specific T cell columns. Standard in vitro phytohemagglutinin (PHA) stimulation of lymphocyte cultures was done with culture media containing 10% SCD serum, as compared to normal pooled O, Rh+ (O+) serum. Mitogenic responses were expressed as mean counts per minute (cpm) and stimulation index of triplicate cultures. Results revealed PHA responses were positive in all experiments when a standard stimulation index of 10 or greater was used as a test parameter for comparison. Positive results were demonstrated in 43/43 (100%) of triplicate cultures regardless of serum type in all experiments. Conversely, by using mean cpm as the test criterion, suppression of PHA response was shown in SCD serum supplemented cells as follow; 36/43 (84%) of PBMC, 35/43 (81%) of CD3 and 37/43 (86%) of CD4 cultures. The degree of suppression ranged from > 10% to 98% in individual experiments, as compared to O+ serum. Inhibitors of normal T lymphocyte in vitro PHA response appear to be present in a significant percentage of SCD sera even during the healthy state of disease. Type 2 cytokines which suppress cell mediated immunity would seem to be the most likely inhibitory agents. PMID- 9399100 TI - Immunoglobulins for intravenous use inhibit TNF alpha cytotoxicity in vitro. AB - Intravenous immunoglobulins (IVIg) have been used as an immunomodulatory therapy in a variety of diseases. Several mechanisms of action have been proposed, one of which is interference with the cytokine network. We have investigated the effect of IVIg on the cytotoxicity of human TNF alpha. IVIg was capable of protecting L929 fibroblasts from TNF alpha induced cell death. This effect was not species specific and was mediated by both the Fc and the Fab portion of immunoglobulins. Since the effect was also seen when IVIg was added after the removal of TNF alpha from the culture medium, it seems to be independent of the interaction of TNF alpha with its receptor. We conclude that IVIg either act on some point of the TNF alpha signalling pathway or influence the cell cycle unspecifically. The cytoprotective effect of IVIg potentially could contribute to the beneficial effect described for various diseases. PMID- 9399101 TI - Amaranthus leucocarpus lectin recognizes human naive T cell subpopulations. AB - Amaranthus leucocarpus lectin (ALL) is specific for GalNAc residue found in the inner core of Gal beta 1,3GalNAc alpha 1,O-Ser/Thr disaccharide (T-antigen) or GalNAc alpha 1,O-Ser/Thr (Tn-antigen). Flow cytometric analysis using fluorescein labeled lectin and monoclonal antibodies against human cell surface markers indicated that 5.7% of mononuclear cells from human healthy donors are recognized by ALL. These cells have the phenotype CD2+CD4+CD19- and most of the lymphocytes recognized are also CD27+, CD45RA+ and CD43+. ALL possesses mitogenic activity on lymphocytes after neuraminidase treatment. Our results indicate that the receptors recognized by ALL could be considered surface markers for naive human T lymphocyte subsets. PMID- 9399102 TI - Early recipient-donor switch of the complement type after liver xenotransplantation. AB - Liver transplantation is an immunological peculiarity with respect to the resistance of the graft to humoral rejection. We undertook a kinetic analysis of molecules involved in humoral rejection for a period of one week following xenografting in the hamster to rat model system. A complement-dependent lymphocytotoxicity test (CDC) was used to detect anti-donor antibodies in the recipient rats. Complement was studied by two methods. Function of the classical complement pathway was evaluated with a hemolytic assay, and C3 was measured by radial immunodiffusion. Conversion of the major plasma proteins from recipient to donor profile was studied by zone electrophoresis on agarose. CDC showed antibody titers rose during the first week post-transplantation, and they were of complement-activating isotypes. Zone electrophoresis showed almost complete replacement of rat C3 by hamster C3 within 72 hours. Hemolytic assay of complement on day 6 post-transplant showed serum of the xenograft recipients could lyse erythrocytes sensitized with rat antibody with 80% of efficiency of normal rat serum. Our data show the effector molecules for humoral rejection, rat antibodies with anti-hamster specificity and a functional complement cascade, were present within the first week following transplantation. Rapid conversion of serum complement to hamster proteins maintains compatibility with the species specific membrane inhibitors of complement activation expressed by the xenografted hepatocytes, and could limit complement-mediated damage. PMID- 9399103 TI - Regulatory role of CD8 in major histocompatibility complex-unrestricted tumoricidal activity of mouse T cells activated with anti-CD3 monoclonal antibody. AB - Antigen-nonspecific CD8+ cytotoxic T cells induced with anti-CD3 monoclonal antibody (mAb) are able to kill tumor cells in a major histocompatibility complex (MHC)-unrestricted fashion. However, the role of CD8 in the MHC-independent tumoricidal activity of anti-CD3-activated killer T (AK-T) cells has not been investigated. Here we show that anti-CD8 alpha mAb inhibits, in a dose-dependent fashion, lysis of P815 and YAC-1 tumor cells by mouse AK-T cells. The inhibition of MHC-unrestricted cytotoxicity by anti-CD8 alpha mAb cannot be attributed to interference with an adhesion-like function of CD8 towards class I MHC molecules on the target cells because anti-CD8 alpha mAb (i) had equal inhibitory effects on the cytolysis of tumor target cells regardless of their relative level of class I MHC molecule expression and (ii) did not interfere with the formation of conjugates between AK-T cells and class I MHC-bearing P815 tumor cells. However, anti-CD8 alpha mAb abrogated AK-T cell granule exocytosis in the presence of P815 tumor cells, indicating a regulatory role for CD8 in the signal transduction events which result in lysis of the tumor target cells. Immunoblot analysis of the post-nuclear fraction of lysates from AK-T cells exposed to P815 tumor cells in the presence of anti-CD8 alpha mAb revealed reduced phosphorylation of tyrosine residues on a protein with an Mr of approximately 62 kDa. Taken together, these data suggest that CD8 is able to affect the tumoricidal activity of MHC-unrestricted AK-T cells independent of class I MHC molecules on the target cell. PMID- 9399104 TI - Mechanisms involved in activation of human eosinophil exocytosis by substance P: an in vitro model of sensory neuroimmunomodulation. AB - Substance P (SP), a tachykinin with a wide range of biological activities including a priming effect on human eosinophil chemotaxis, was investigated for its influence on eosinophil cytotoxic function measured as degranulation of eosinophil-derived neurotoxin (EDN). Peripheral blood was obtained from healthy volunteers and the degranulation assays were performed using radioimmunoassay (RIA). SP and its C-terminal elicited EDN release in a time-dependent mode at a narrow range of doses with optimal activity of 10(-6) M. FK888 (NK-1 receptor antagonist) inhibited EDN release stimulated by SP in dose dependency, also a complete inhibition was observed when eosinophils were preincubated with 1000 ng/ml pertussis toxin (PTX). Pre-exposure of eosinophils to staurosporine resulted in blockage of SP-induced EDN release in a dose-dependent mode. On the other hand, SP at 10(-7) M and 10(-8) M primed eosinophils to suboptimal dose (10(-8) M) of Platelet activating factor (PAF) resulting into significant enhancement of EDN release. SP(4-11) fragment showed a similar activity while SP(1-4) fragment was not active. SP priming of eosinophils was not affected by Ca2+ depletion, however, it caused a change in the pattern of the intracellular calcium influx against the suboptimal dose of PAF. These results suggest that SP i) may induced human eosinophil matrix protein degranulation through a receptor mediated mechanism coupled to PTX sensitive G protein(s) with the probability of linkage to phospholipase C activation, and, ii) primes human eosinophils for an exalted inflammatory response through a Ca2+ independent pathway. PMID- 9399105 TI - CD5+ B cell-dependent regulation of the murine T-cell independent immune response against the human blood group A antigen. AB - The CD5+B lymphocyte (B1a) population is known to be involved in most immune responses to microorganism TI antigens. Moreover, xid mice deficient for immune responses against TI-2 antigens are known to lack the B1a population, suggesting a role for B1a cells in TI-2 immune responses. We previously established that the oligosaccharide human blood group A antigen stimulated murine TI-2 immune responses. In this work, we show that the frequency of anti-A-secreting hybridomas was higher in mice with larger splenic B1a populations and that in vivo anti-CD5 treatment reduced anti-A immune response without affecting the response against TD RBC antigens. A similar effect was observed by in vitro anti CD5 treatment of splenocytes. The in vivo anti-CD5 treatment also interfered with the immunization-dependent increase in splenocyte numbers. These results are in agreement with an important role for the B-cell CD5 receptor in the regulation of TI-2 immune responses possibly mediated by its interaction with the CD72 ligand. PMID- 9399106 TI - Interleukin-1 and interleukin-1 receptor antagonist in systemic lupus erythematosus. AB - Interleukin-1 (IL-1) is thought to play an important role in the immunopathology of systemic lupus erythematosus (SLE). IL-1 receptor antagonist (IL-1ra) exhibits a dose-responsive inhibition of IL-1 effects, and Fcr receptors play a key role in IL-1ra production. To clarify the relationship between IL-1, IL-1 receptor antagonist (IL-1ra) production, and Fcr receptors in SLE, fifteen untreated lupus patients (9 females and 6 males) were evaluated. IL-1 and IL-1ra production from both monocytes and neutrophils was determined by both a murine thymocyte proliferation assay and ELISA. The expression of Fcr receptors on both monocytes and polymorphonuclear cells was measured by flow cytometry. There was no significant difference between IL-1 beta and IL-1ra production from ex vivo monocytes between lupus patients and normals. However, serum IL-1ra was significantly higher in lupus patients than in normals. The expression of FcrRII (CD32) on monocytes was lower in lupus patients than in normals. There was no correlation between the expression of FcrRII and the serum immune complexes; the production of IL-1ra and the expression of CD32, serum immune complex levels, and serum IgG. Both serum IL-ra and the production of IL-1ra had no correlation to SLEDAI, C3, C4, C1q, or ESR. These observations suggest that although IL-1 and IL 1ra may not play a major role in the initiation of pathogenesis of lupus, the low FcrRII expression in lupus may reflect low immune complex clearance and contribute to disease pathogenesis. PMID- 9399107 TI - Immunoassay of human Neisseria meningitidis serogroup A antibody. AB - An enzyme immunoassay is described which quantitates antibodies to Neisseria meningitidis serogroup A capsular polysaccharide in human sera. Modifications of a previously developed two-day assay by Carlone et al. were made to permit analysis in one day and to be compatible with automation. The allowable variations in assay conditions and the areas in which stringent control must be maintained for consistent assay performance are described. Antigen-coating parameters, the kinetics of primary and secondary antibody incubation steps, the buffer compositions, including detergents, serum requirements, and the need for blocking steps were examined. Our modified one-day assay showed excellent agreement with the standardized method of Carlone, with a correlation coefficient between the two methods of 0.989. This assay is adaptable within a permissible range of parameters thus facilitating the implementation of the standardized assay. This will maximize the consistency of results from serum analysis for conjugate vaccine trials related to serotype A Neisseria meningitidis. PMID- 9399108 TI - Gender, race, class, and aging: advances and opportunities. AB - Key debates in social science and health research have centered on how to increase the inclusiveness of such research and hence its relevance for understanding the intersections of race, class, gender, and aging. This article uses gerontology as a case in point, examining the challenges of inclusivity and interlocking oppressions/intersectionality for better apprehending how broad structural factors shape and determine the experience of aging and growing old. The authors discuss alternative hypotheses being used to explore inequalities in the aging experience and the limitations of current concepts and methods. Promising new developments in sociology, epidemiology, and other fields are described in terms of their relevance for better understanding the dynamic interplay of race, class, gender, and aging. PMID- 9399109 TI - The potential role of unemployment benefits in shaping the mental health impact of unemployment. AB - This study looks at the association between formal systems of support (unemployment compensation or welfare) and mental health outcomes during periods of unemployment. It assesses whether unemployed persons not receiving unemployment benefits are at greater risk of reporting depression and suffering ill-health than those receiving some kind of unemployment compensation, independent of total household income. The authors performed a secondary analysis of data collected in the National Survey of Families and Households, 1987-1988. Outcome measures included an index of depression and perception of health status. Multiple regression analyses were used. The unemployed receiving unemployment compensation or benefits from other entitlement programs did not report significantly higher depression relative to the employed. Rather, the study found a significantly higher index of depression among unemployed persons receiving welfare benefits or no benefits, even after controlling for total household income and previous employment/unemployment history. Thus unemployment compensation may play an important role in ameliorating the impact of unemployment on depression. By contrast, means-tested benefits may not be sufficient to reduce the risk for reporting depression and suffering ill-health in comparison with the full-time employed. The implications of the findings are discussed in terms of social policy development. PMID- 9399110 TI - Karoshi--death from overwork: occupational health consequences of Japanese production management. AB - There is considerable international interest in Japanese production management (JPM), known in the West as "lean production." Advocates of this new form of management argue that it improves both economic productivity and health. In Japan, however, the relationship between JPM and sudden death due to cardiovascular and cerebrovascular disease has been an important topic of debate since the 1970s. Japanese have named these types of deaths karoshi, which means "death from overwork." In North America and Western Europe a number of studies have demonstrated a significant relationship between high job strain (high production demands and low levels of control and social support) and cardiovascular disease. This article reviews the elements of JPM and examines their potential health consequences. The authors present an overview of karoshi, discuss its possible connections to specific ideological and organizational characteristics of JPM, and suggest the job strain mechanism as a possible pathway between karoshi and JPM. They conclude by discussing the need for comparative research that examines the health effects of work organization and management methods cross-culturally. PMID- 9399111 TI - The growth of corporate private hospitals in Malaysia: policy contradictions in health system pluralism. AB - The rapid growth of corporate investment in the Malaysian private hospital sector has had a considerable impact on the health care system. Sustained economic growth, the development of new urban areas, an enlarged middle class, and the inclusion of hospital insurance in salary packages have all contributed to a financially lucrative investment environment for hospital entrepreneurs. Many of Malaysia's most technologically advanced hospitals employing leading specialists are owned and operated as corporate business ventures. Corporate hospital investment has been actively encouraged by the government, which regards an expanded private sector as a vital complement to the public hospital system. Yet this rapid growth of corporately owned private hospitals has posed serious contradictions for health care policy in terms of issues such as equity, cost and quality, the effect on the wider health system, and the very role of the state in health care provision. This article describes the growth of corporate investment in Malaysia's private hospital sector and explores some of the attendant policy contradictions. PMID- 9399112 TI - Caught in the "traps of managed competition"? Examples of Russian health care reforms from St. Petersburg and the Leningrad region. AB - Elements of a "managed market" for health services have been introduced into the Russian health care system, which under the Soviet regime was run as a comprehensive state-managed system. The authors examine the recent development of health service reforms in a case study of the city of St. Petersburg and the surrounding Leningrad region. Evidence from key informants and a local survey of service users shows how alternative models of the managed market are being introduced in different parts of the study area. A critical review of the market oriented strategies for reform emerging in the case study suggests that such reforms carry risks associated with the "traps of managed competition." Future policy for health service systems in Russia must take these risks more fully into account. PMID- 9399113 TI - The future of primary care in a managed care era. AB - Health care reform in the United States and elsewhere raises many questions about equity and effectiveness of health services. Although the impetus has been cost containment, the reforms have often been justified on the grounds that they will enhance primary care. In this article, health care reform efforts are divided into two types: market-driven, demand-based systems versus systems predicated on meeting population health needs. The two "scenarios" are contrasted with regard to their likely impact on the attainment of primary care characteristics: first contact care, longitudinality, comprehensive services, and coordination. Since the ultimate outcome of these reforms cannot be predicted, there is compelling need for evaluating them as they proceed. PMID- 9399114 TI - Local government decision-making and access to primary physician services in Norway. AB - Public responsibility for health care can be justified by ambitious egalitarian objectives, as it is commonly believed that the private sector generates greater disparities than the public sector. Government institutions can be designed to achieve equality in provision of health services. The article addresses the geographical distribution of primary care physicians in Norway, where primary physician services are the responsibility of local governments, primarily financed by general taxation. The authors analyze the allocation of physicians using a local government demand model, a synthesis of consumers' demand and local government resource allocation. Analyses were performed on a panel data set of all Norwegian municipalities covering the period 1986-1992. The results are encouraging. A decentralized system of primary physician services does seem to be fairly effective in securing equity in access to these services for the municipal population. In particular, local governments seem to respond well to the health care needs of their populations. Distribution of physicians is only to a very small extent dependent on the wealth of the municipality. PMID- 9399116 TI - Special interests or citizens' rights? "Senior power," Social Security, and Medicare. AB - Conventional political analysts and mainstream media accounts attribute substantial political power to the elderly in the United States. This attribution of "senior power" is usually made in the context of the politics of Social Security and Medicare. This article contrasts the conventional construction of elderly political actors as a special interest with a more critical perspective that views Social Security and Medicare as citizens' rights. Critical examination of the welfare state's role in creating age as a potential political cleavage and the politics of Social Security and Medicare reveals that there is no undifferentiated politics of aging in the United States. Rather, age interacts with a variety of other statuses such as race/ethnicity, gender, and class to condition citizens' political mobilization. Welfare state policies--social insurance programs like Social Security and Medicare, means--tested programs like Medicaid and Supplemental Security Income, and targeted tax expenditures for private pensions and health insurance--differentially empower particular subgroups of elderly citizens and routinely disadvantage the most vulnerable elderly, including minority elders, women, and the oldest old. PMID- 9399115 TI - The challenge of moving from welfare to work. AB - Despite overall low unemployment rates in 1996, various subgroups of the population had rates much higher than the average. This brief article considers the labor market profile of welfare recipients and examines the unemployment and underemployment rates of groups with similar demographic characteristics. In some cases, such rates are three and four times the overall national average. PMID- 9399117 TI - Austria's new attendance allowance: a consumer-choice model of care for the frail and disabled. AB - Austria's new social welfare provision, an attendance allowance, adopted in 1993, is a prototype for a consumer-directed care model for persons who are frail or disabled. This article describes the background to and provisions of the new legislation, including the unique political interests from the left and the right that converged to facilitate its passage. The author discusses eligibility criteria, the consumer autonomy principles underlying the model, and the impacts and implications anticipated. Such a model has been mentioned as an option for states considering creative expansion of long-term care provisions in the United States. PMID- 9399118 TI - The use of mass campaigns in the expanded program on immunization: a review of reported advantages and disadvantages. AB - The use of mass immunization campaigns (MICs) has been and remains controversial. To evaluate these campaigns, the authors review the literature relating to their effectiveness, sustainability, and cost-effectiveness in controlling diseases and raising immunization coverage levels, and their impact on the subsequent development of routine immunization services. Well-conducted campaigns have increased vaccine coverage levels and decreased disease morbidity and mortality. Their use in the Americas has been associated with the apparent elimination of poliomyelitis. However, unless health care infrastructure is improved, or campaigns are repeated, gains in coverage levels may not be sustained. Studies suggest that MICs are often not as cost-effective for raising coverage as the delivery of vaccines through routine services, but the use of coverage as the only outcome measure is questionable. Mass immunization campaigns can increase awareness of vaccination and may be appropriate in situations where new programs are to be initiated, in refugee situations where people congregate into areas with little infrastructure, and in disease eradication efforts when specific time goals are set. Little information is available on whether MICs strengthen or interfere with the development of routine services. To be successful, MICs require a well-coordinated and planned effort on the part of national authorities with the identification of specific goals, intensive social promotion, and strong management. In addition, research is needed to clarify how MICs should be evaluated. PMID- 9399119 TI - Economy, politics, and health status in Cuba. AB - An economic contraction occurred in Cuba at the beginning of the 1990s, of a magnitude greater than in any developed country in the last half century. This resulted primarily from the disappearance of the European socialist bloc and simultaneous tightening of the U.S. government's blockade at a time when Cuba was engaged in correcting its main economic problems. The economic crisis affected a number of areas of Cuban society. The state adopted a series of measures to cope with the socioeconomic situation, which have yielded positive results in the social and economic fields, as well as some undesirable results. In the health sector, the economic crisis has mainly reduced the availability of resources and has adversely affected some health determinants and some aspects of the population's health status. Despite the prevailing economic difficulties, the government is determined to preserve the country's achievements in health, and to develop them still further. The solution is not privatization or the introduction of health insurance systems or similar measures. Rather, Cuba will seek greater rationality and economic efficiency in the health sector. It has ratified the principles that the state should continue to finance the health system and maintain universal coverage and accessibility through free services. PMID- 9399120 TI - The analysis of EDTA in dried bloodstains by electrospray LC-MS-MS and ion chromatography. AB - Analytical methods were developed to determine the presence of ethylenediaminetetraacetic acid (EDTA) in dried bloodstains to provide probative information when allegations of evidence tampering have been made in criminal cases. A simple screening method using ion chromatography to analyze stains was found to be quantitative to the 5 ppm level. The presence of EDTA was then confirmed using negative and positive ion mode liquid chromatography-tandem mass spectrometry (LC-MS-MS) methods. A blind trial of these methods on 42 samples correctly determined the bloodstains that did and did not contain the preservative EDTA. One interesting observation in these results was the adsorption and postanalysis release of EDTA in the chromatographic system. In order to avoid cross contamination of samples resulting from this phenomena, it was found to be necessary to use EDTA-free blood extracts as blanks in the LC-MS analysis of bloodstains. PMID- 9399121 TI - Identification of tramadol and its metabolites in blood from drug-related deaths and drug-impaired drivers. AB - Tramadol is a centrally acting, binary analgesic that is neither an opiate derived nor a nonsteroidal anti-inflammatory drug and that was approved for use in the United States in 1995. It is used to control moderate pain in chronic pain settings such as osteoarthritis and postoperative cases. Used in therapy as a racemic mixture, the (+)-enantiomer weakly binds to the mu-opioid receptor, and both enantiomers inhibit serotonin and norepinephrine reuptake. Tramadol's major active metabolite, O-desmethyltramadol (ODT), shows higher affinity for the mu opioid receptor and has twice the analgesic potency of the parent drug. The synergism of these effects contributes to tramadol's analgesic properties with the (+)-enantiomer exhibiting 10-fold higher analgesic activity than the (-) enantiomer. Although tramadol was initially thought to exhibit low abuse potential, Ortho-McNeil, the drug's manufacturer, recently reported a large number of adverse events attributed to tramadol including abuse by opioid dependent patients, allergic reactions, and seizures. The high number of adverse reactions has prompted the company to update the prescribing information for the drug. An analytical method using gas chromatography-mass spectrometry (GC-MS) without derivatization for the determination of tramadol and its metabolites is reported. An n-butyl chloride extraction is followed by GC-MS analysis using a 5% phenylmethylsilicone column (30 m x 0.32-micron i.d.). Analysis of 12 blood samples from tramadol-related deaths and four nonfatal intoxications involving tramadol revealed concentrations ranging from 0.03 to 22.59 mg/L for tramadol, from 0.02 to 1.84 mg/L for ODT, and from 0.01 to 2.08 mg/L for N desmethyltramadol. Three deaths were clearly attributable to acute morphine toxicity, one was a doxepin overdose, and six were multiple drug overdoses. The role of tramadol in each death is explored. PMID- 9399122 TI - GC-MS-MS confirmation of unusually high delta 9-tetrahydrocannabinol levels in two postmortem blood samples. AB - Unusually high levels of delta 9-tetrahydrocannabinol (delta 9-THC) were detected in two postmortem blood samples. Because of sample decomposition, the major metabolite, 11-nor-9-carboxy-tetrahydrocannabinol (delta 9-THCCOOH), could not be determined using the routine EI-MS technique, which cast some doubt on the delta 9-THC result. Analysis of the sample extracts by GC-MS-MS confirmed the presence of delta 9-THC, although no delta 9-THCCOOH was detected. PMID- 9399123 TI - Multicomponent spectroscopic assay for hemoglobin and ferrihemoglobin species in methemoglobin treatment of cyanide poisoning. AB - Monitoring the concentrations of various hemoglobin and ferrihemoglobin species and their cyanide complexes is important in the study of the efficacy of methemoglobin-forming agents for the treatment of cyanide toxicity. In this study, the visible absorption spectra of three hemoglobin intermediates were experimentally determined and compared with computer-generated spectra. The data supported the assumption that the molar absorptivities of the intermediates are equivalent to the combined absorptivities of the component subunits. A multicomponent Fourier transform (FT)-aided full spectrum quantitation system (FSQ) to simultaneously measure hemoglobin (Hb), hemimethemoglobin (hemiMetHb), and the cyanide complex dicyanohemimethemoglobin (dicyhemiMetHb) was also evaluated. It was found that FSQ had satisfactory accuracy and precision to quantitate hemiMetHb and dicyhemiMetHb at concentrations from 2 to 20% of the total Hb, a range commonly encountered in the treatment of cyanide poisoning using methemoglobin-forming agents. The simplicity and rapid throughput of the method make it suitable for clinical evaluation studies and form the basis for the design of a portable instrument for field analysis of these species. PMID- 9399124 TI - Reversed-phase HPLC determination of eight anticoagulant rodenticides in animal liver. AB - A reversed-phase high-performance liquid chromatographic method was developed for the analysis of eight anticoagulant rodenticides in animal liver. Coumarinic anticoagulant rodenticides (brodifacoum, bromadiolone, coumachlor, coumatetralyl, difenacoum, and warfarin) were detected by using a gradient elution and a fluorimetric detection. Indanedione anticoagulant rodenticides (chlorophacinone and diphacinone) were detected by using an isocratic elution and an UV detection. Anticoagulants were extracted from liver with mixtures of acetone/diethylether and acetone/chloroform. Extracts were applied to solid-phase extraction cartridges. Linearity was checked over the concentration range 0.1-0.6 microgram/g. Relative standard deviations of within-run and between-run variability were all between 5.7 and 10.3%. Recoveries from spiked liver samples were between 51.7 (difenacoum) and 78.2% (warfarin). Limits of detection were between 0.01 (difenacoum and warfarin) and 0.11 microgram/g (chlorophacinone). PMID- 9399125 TI - The online screening technique for urinary benzodiazepines: comparison with EMIT, FPIA, and GC-MS. AB - Three commercial immunoassay systems (EMIT, EPIA, Online) for the screening of benzodiazepines in urine were evaluated using authentic patient samples with gas chromatography-mass spectrometry (GC-MS) as the reference method. The Online system (kinetic interaction of microparticles in solution) gained in performance by applying a 100-ng/mL cutoff limit and by incorporating beta-glucuronidase treatment, which could be automated on the Cobas Mira Plus instrument. When using enzymatic hydrolysis, all three immunoassay systems had high levels of sensitivity, including samples containing only flunitrazepam and nitrazepam metabolites. A high degree of concordance was observed between the Online and FPIA (fluorescence polarization immunoassay) systems when analyzing 138 randomly selected patient samples. The EMIT II and EMIT d.a.u. (enzyme multiplied immuno technique) systems gave a higher number of positive results, but the presence of benzodiazepines could not be verified by GC-MS in a substantial number of these cases. The rate of unconfirmed positive results was increased when enzyme hydrolysis was incorporated in the EMIT II assay. Although differences in the performances of the investigated assay systems were observed, they all seem appropriate for clinical use in detecting benzodiazepine intake in drug abusers when enzymatic hydrolysis is included. PMID- 9399126 TI - Standardization of a method for the routine analysis of polychlorinated biphenyl congeners and selected pesticides in human serum and milk. AB - A methodology is presented for the routine determination of specific polychlorinated biphenyl (PCB) congeners in serum and milk samples. The procedures include standardized extraction, cleanup and quantitation by high resolution gas chromatography (GC) and comprehensive quality assurance program to minimize systematic and erratic errors. The analyses of 68 PCB congeners and three pesticides, p,p1-dichloro diphenyl dichloro ethylene (DDE), hexachlorobenzene (HCB), and Mirex, at part-per-billion levels include the addition of surrogate congener standards (IUPAC isomers #46 and #142), extraction with hexane after protein precipitation, cleanup with Florisil, and analysis by GC with capillary column and electron capture detection. Quantitation is based on calibration standards and response factors using isomers #30 and #204 as internal standards. The quality control activities consist of analyses of samples in batches of 6 to 10 simultaneously with quality control (QC) samples. The quality assurance program checks that the procedures are under control by the use of control charts and set the criteria for data acceptability. The detection limits for the congeners and pesticides associated with the analyses of 500 serum samples and of 100 milk samples are reported. In addition, typical profiles of congener distribution in both matrices are illustrated. PMID- 9399127 TI - A fatality involving zolpidem. AB - An elderly woman residing in an independent-living retirement community was found dead in the bathtub. She had a history of depression and was prescribed Ambien (generic name zolpidem) for the treatment of insomnia. Two empty prescription bottles of Ambien were found; both were for 10-mg tablets with a total quantity of 60. A postmortem examination was conducted, and blood, urine, and gastric contents were submitted for toxicology screening. The cause of death was drowning. The only remarkable toxicology finding was zolpidem. Quantitative analysis by gas chromatography-mass spectrometry determined the following concentrations of zolpidem: blood, 7.9 micrograms/mL and urine, 4.1 micrograms/mL. A total of 7 mg unabsorbed zolpidem was found in the gastric contents. Our findings report the highest blood concentration of zolpidem reported to date and corroborate other studies that imply that death due solely to overdosage of zolpidem is an unlikely occurrence. PMID- 9399128 TI - Excretion of MBDB and BDB in urine, saliva, and sweat following single oral administration. AB - A procedure based on gas chromatography-mass spectrometry (GC-MS) for the simultaneous identification of N-methyl-1-(3,4-methylenedioxyphenyl)-2-butanamine (MBDB) and its desmethylated metabolite 3,4-(methylenedioxyphenyl)-2-butanamine (BDB) in urine, saliva, and sweat specimens is presented. For urine and saliva, the method involved the alkaline extraction of a 1-mL specimen (MDEA-d5 internal standard) into ethyl acetate followed by derivatization with heptafluorobutyric anhydride. Sweat specimens, which were collected by a sweat patch, were tested after methanolic elution. The procedure was used to study the excretion of MBDB and BDB in urine, saliva, and sweat after single oral administration of 100 mg of MBDB to one subject. Urine tested positive for 36 h with a peak concentration at 4 h. Immunoassays were positive for 24 and 4 h using FPIA and EMIT, respectively. Peak saliva concentration was observed at 2 h. MBDB and BDB were detected in saliva during the first 17 h. Finally, both compounds were excreted into sweat with a constant increase in concentration during the first 36 h followed by a decrease for the remaining time. In all the biological specimens that were tested, MBDB was present in higher concentrations than its metabolite. PMID- 9399129 TI - Diltiazem and pentoxifylline determination in postmortem specimens. AB - A 78-year-old woman was found dead in her basement. Qualitative screening of available postmortem specimens detected the presence of diltiazem and pentoxifylline. Quantitations were carried out by gas chromatography using nitrogen-phosphorus detection and confirmed by gas chromatography-mass spectrometry with the following results: blood, 0.59 mg/dL diltiazem and 0.63 mg/dL pentoxifylline; urine, 1.17 mg/dL diltiazem and 0.08 mg/dL pentoxifylline; bile, 0.40 mg/dL diltiazem and 0.22 mg/dL pentoxifylline; gastric contents, 0.28 mg/dL diltiazem and 0.02 mg/dL pentoxifylline. Both drugs were found qualitatively in formaline-fixed tissues. PMID- 9399130 TI - Analytical findings in a fatal poisoning with silver compound. AB - A 69-year-old male radio technician drank an unknown amount of a liquid and died within 5 h at a hospital. Mainly silver, potassium, and calcium, among other substances, were found in the residue of the liquid from the empty bottle by means of the spectrographic method (30.7% silver and 25.5% potassium using atomic absorption spectometry [AAS] method). The toxicological analysis of the postmortem material for silver performed by the flame AAS method (stomach, 2.43 micrograms/g; intestines, 1.12 micrograms/g; liver, 6.29 micrograms/g; kidney, 4.85 micrograms/g; spleen, 30.1 micrograms/g; heart, 10.8 micrograms/g; lung, 14.8 micrograms/g; and brain, 0.61 microgram/g) confirmed fatal silver compound poisoning. The results were verified by the standard additions technique and recovery examination. However, no increase in the potassium concentrations was observed in the postmortem material. There have been no data in available literature on the distribution of silver in tissues in people after oral administration of silver salts. PMID- 9399132 TI - Appreciable blood-ethanol concentration after washing abraised and lacerated skin with surgical spirit. PMID- 9399131 TI - A case of homicidal poisoning involving several drugs. AB - Accidental or suicidal poisonings due to benzodiazepines have been previously reported. A case of fatal, homicidal poisoning with benzodiazepines, antipyretic analgesics (anti-inflammatory drugs), and beer is described here. In this homicidal poisoning, the drugs and beer were given to the decedent by his wife. Autopsy findings showed no clinically significant macroscopic findings except for slight postmortem change. Capillary gas chromatography coupled to mass spectrometry was employed to quantitate the drugs in biological fluids and stomach contents. Six drugs (brotizolam, triazolam, ibuprofen, dihydrocodeine, phenylpropanolamine, and chlorpheniramine) were identified and quantitated in blood, urine, and stomach contents. Although each drug was present in a very small quantity, the cause of death was determined to be the combination of these drugs and alcohol poisoning. PMID- 9399133 TI - Immunoassay responses of MBDB. PMID- 9399134 TI - Ecstasy and related substances--serum levels in impaired drivers. PMID- 9399135 TI - Face facts: a history of physiognomy from ancient Mesopotamia to the end of the 19th century. AB - Inscribed on the face is a code, the translation of which has entertained and eluded humankind for many centuries. The practice of reading the face dates back as early as the paleobabylonian period in Mesopotamia. It wasn't until much later, however, that this ancient tradition was named. PMID- 9399136 TI - Bringing it into focus: visual cues and their role in directing attention. AB - Visual cues are powerful tools for directing the user's eyes to relevant information. When incorporated into the design of a medical/scientific illustration, important details are more quickly conveyed. This article discusses the use of line, color, contrast, light and shadow, spatial relationships, image size and location, rhythm, and graphic devices as attention-directing cues utilized in focusing the user's eyes to pertinent information. PMID- 9399138 TI - The use of biomaterials in the repair of abdominal wall defects: a comparative study between polypropylene meshes (Marlex) and a new polytetrafluoroethylene prosthesis (Dual Mesh). AB - In this study we compared the behaviour of the non-porous on one side ePTFE Dual Mesh prosthesis and the macroporous polypropylene mesh Marlex in the repair of abdominal wall defects in rabbits. We evaluated the degree of integration with recipient tissue, biological tolerance, adhesion formation with viscera and the biomechanical resistance of the repair zone. Our results showed good biological tolerance of both prostheses and a high degree of adhesion formation in Marlex implants. In animals with Dual Mesh implants, only loose adhesions were seen. Marlex implants induced the presence of disorganized scar tissue, while the Dual Mesh prostheses were encapsulated by organized tissue. The macrophage response was similar in both decreasing with time. The resistance to traction was higher when the reparation was done with polypropylene. We concluded that the structure of the prosthesis determines its degree of integration and the resistance to traction of the repaired zone. PMID- 9399137 TI - Chemical and physical characterization of a novel poly(carbonate urea) urethane surface with protein crosslinker sites. AB - A major complication which occurs with implantable polyurethane biomaterials is bioincompatibility between blood and the biomaterial surface. Development of a novel biodurable polyurethane surface to which biological agents, such as growth factors or anticoagulants could be covalently bound, would be beneficial. The purpose of this study was to synthesize a novel poly(carbonate urea) urethane polymer with carboxylic acid groups which would serve as "anchor" sites for protein attachment. Physical characteristics such as tensile strength, initial modulus, ultimate elongation, tear strength, water/alcohol uptake and water vapor permeation were then evaluated and compared to other biomedical-grade polyurethanes. Covalent linkage of the blood protein albumin to this novel surface was then examined. A biodurable polycarbonate-based polyurethane containing carboxylic acid groups (cPU) was synthesized using a two step procedure incorporating the chain extender 2,2-bis(hydroxymethyl)-propionic acid (DHMPA). Tensile strength of this cPU film was 2.7 and 2.6 fold greater than both a polycarbonate-based polyurethane synthesized with a 1,4-butanediol chain extender (bdPU) and Mitrathane (Mit) controls, respectively. The cPU polymer also possessed 7.8 and 31 fold greater structural rigidity upon evaluation of initial modulus as compared to the bdPU and Mit, respectively. Ultimate elongation for the bdPU films was slightly higher than the cPU and Mit films, which had comparable elongation properties. The force required to tear the bdPU film was 1.9 and 32 fold greater than the cPU and Mit films, respectively. Alcohol solution uptake by all of the polyurethane segments increased with increasing alcohol concentrations, with the cPU having the greatest uptake. Water uptake was minimal for all the polyurethanes examined and was not affected by altering pH. Water vapor permeation was lowest for the cPU films as compared to both bdPU and Mit. Swelling the cPU in 50% ethanol prior to evaluation slightly increased water vapor permeation through the films. Covalent linkage of the radiolabelled blood protein albumin (125I-BSA) to the cPU segments incubated with the heterobifunctional crosslinker 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) was greatest in the higher percent of ethanol as compared to controls. These results serve as foundation for developing a novel poly(carbonate urea) urethane with physical characteristics comparable to other medical-grade polyurethanes while having protein binding capabilities. PMID- 9399139 TI - Assessment of encrustation behaviour on urinary tract biomaterials. AB - The effective clinical use of biomaterials within the urinary tract is often hindered by the associated problems of bacterial biofilm formation and encrustation which may cause obstruction or blockage of urethral catheters and ureteral stents. Methods for assessing encrustation formation on these devices are reviewed and novel urinary tract biomaterials which may be more effective at resisting encrustation are discussed. PMID- 9399140 TI - Blood compatible phospholipid-containing polyurethanes: synthesis characterization and blood compatibility evaluation. AB - A new diol, bis[2-(2-hydroxyethyldimethylammonio)ethyl] butylenediphosphate, that contains two phosphatidylcholine analogous moieties in one diol molecule, was synthesized and characterized. The diol together with 1,4-butanediol as chain extender was further incorporated into the propolymer of poly(ethylene oxide) (Mn = 1000, 2000, 6000) and 4,4'-methylenediphenyl diisocyanate. The resulting phospholipid poly(ether urethane)s show viscosity behavior of common polyelectrolytes. The bulk and surface characteristics of the new phospholipid poly(ether urethane)s were investigated by IR, GPC, ATR-FTIR, ESCA and contact angle measurements. The new polymers possessed relatively hydrophilic surface revealed by contact angle measurements. The blood compatibilities of the polyurethanes were evaluated by platelet rich plasma contacting studies and scanning electron microscopy observation using medical grade PVC as the reference. No platelet adhesion was observed for all new phospholipid polyurethane casting films. This new type of phospholipid polyurethane is expected to have potential biomedical applications. PMID- 9399141 TI - Synthesis, characterization, biodegradation, and drug delivery application of biodegradable lactic/glycolic acid oligomers: I. Synthesis and characterization. AB - A series of oligomers or low molecular weight polymers of lactic and/or glycolic acid has been synthesized with different molar ratios of lactic to glycolic acid. These oligomers have been characterized with respect to oligomer composition, molecular weight, intrinsic viscosity, crystallinity, melting temperature, and glass transition temperature. The polymerization conditions for the lactic/glycolic acid oligomer syntheses were as follows: 180-220 degrees C, 5 mm Hg, 5 h, and 0.1 wt% of catalyst (antimony oxide) concentration. The polymeric compositions correlated to the feed ratios of lactic to glycolic acid. The molecular weight of the oligomers ranged from 895.8 +/- 48.7 to 1368.0 +/- 0 D with the intrinsic viscosity ranging from 0.0513 to 0.0814 dl g-1. The lactic/glycolic acid oligomers were found to be amorphous. The glass transition temperatures of the lactic/glycolic acid oligomers were lower than physiological temperature. PMID- 9399142 TI - New treatments using alginate in order to reduce the calcification of bovine bioprosthetic heart valve tissue. AB - Calcification limits the functional lifetime of cardiac valve substitutes fabricated from glutaraldehyde preserved bovine pericardium. Host factors, mainly younger age, and implant factors, mainly glutaraldehyde cross-linking, are implicated in the calcification process. Glutaraldehyde cross-linking is believed to activate the potential sites in the tissues for biocalcification. In the present work, we investigated the possibility of using alginate azide (AA) instead of glutaraldehyde for the preservation of pericardial tissues in order to enhance the durability of bioprosthetic heart valves. Grafting with poly(GMA-BA) copolymer to the alginate azide cross-linked pericardial (AACPC) tissue was carried out to obtain better stability, strength, and anticalcification properties. The strength property and thermal stability of the AA cross-linked tissues were studied. Calcification studies in rat subdermal models reveal that AA cross-linking reduces the calcification to negligible levels. After 30 days implantation, the calcium content was found to be 10.4 +/- 1.2 and 6.1 +/- 0.3 micrograms mg-1 for untreated AACPC and polymer grafted AACPC, respectively, compared to a value of 100 +/- 1.2 micrograms mg-1 calcium recorded for control glutaraldehyde cross-linked pericardial (GCPC) tissues. PMID- 9399143 TI - Design of macromolecular biological response modifier by immobilizing of D glucose analogue of muramyl dipeptide on carboxymethyl-dextran having mannose branches. AB - It is well known that muramyl dipeptide is a minimum required structure of bacterial peptidoglycan responsible for immunoadjuvant activity. Since mannose receptors exist on the surface of macrophages, polymers with branched mannose residues are expected to target moieties to macrophages. To achieve an efficient delivery of D-glucose analogue of muramyl dipeptide (GADP) via receptor-mediated endocytosis by mannose receptors on the surface of macrophages, GADP/carboxymethyl-dextran (CM-Dex)/Man conjugate was synthesized. Moreover, to study the effect of the introduction of mannose residues, we also synthesized GADP/CM-glucomannan (CM-GM) and GADP/CM-Dex conjugates. The immunological enhancement activities of their conjugates were evaluated by measurements of glucose consumption and beta-D-glucuronidase activity from macrophage-like cells. The GADP/CM-Dex/Man and GADP/CM-GM conjugates showed higher immunological enhancement activity than the GADP/CM-Dex conjugate. The immunological enhancement activity of GADP/CM-Dex/Man and GADP/CM-GM conjugates was decreased to the same level of immunological enhancement activity of GADP/CM-Dex conjugate under the presence of excess mannose. These results suggested that the introduction of mannose residues into GADP/CM-Dex conjugate could increase the affinity against macrophage and the immunological enhancement activity of GADP/CM Dex conjugate itself. PMID- 9399144 TI - Rifampicin carrying polyhydroxybutyrate microspheres as a potential chemoembolization agent. AB - In this study, we attempted to prepare microspheres from a microbial biodegradable polyester, i.e. polyhydroxybutyrate (PHB) as a potential chemoembolization agent. The drug loaded PHB microspheres were prepared by a solvent evaporation technique, in which methylene chloride, distilled water, and polyvinyl alcohol were utilized as the solvent, dispersion medium, and emulsifier, respectively. Microspheres were obtained within a size range of 5-100 microns by changing the initial polymer/solvent ratio, emulsifier concentration, stirring rate, and initial drug concentration. It was possible to obtain PHB with very narrow size distributions by applying gravity field-flow fractionation technique. Very high drug loadings of up to 407.6 mg rifampicin/g PHB were achieved. Drug release rates were very rapid. Almost 90% of the drug loaded was released in about 24 h. Both the size and drug content of PHB microspheres were found to be effective in controlling the drug release from these microspheres. PMID- 9399145 TI - Immobilization of functionalized alkyl-poly(ethylene oxide) surfactants on poly(ethylene) surfaces by means of an argon plasma treatment. AB - Alkyl-poly(ethylene oxide) (PEO) surfactants containing a terminal hydroxyl, sulfate, or carboxylate group were grafted at the surface of poly(ethylene) (PE) samples to improve their blood compatibility. Grafting was achieved by immobilizing PEO surfactants on PE using an argon plasma treatment. The sulfate group containing PEO surfactant was synthesized by sulfating polyoxyethylene(20)stearylether (Brij78; B) with chlorosulfonic acid. A carboxylate-terminated surfactant was synthesized by a substitution reaction of the sodium alkoxide form of B with sodium iodoacetate. XPS analysis of the modified PE samples showed that at short plasma treatment times of up to 5 s the structure of the immobilized surfactants is largely retained. When plasma treatment times longer than 30 s were applied, the PEO chains of the surfactants were degraded. The wettability of the modified PE samples was improved compared to the unmodified PE samples. The wettability of the modified samples did not change when they were stored in air at room temperature for at least 12 weeks. PMID- 9399146 TI - Use of chemically modified nucleotides to determine a 62-nucleotide RNA crystal structure: a survey of phosphorothioates, Br, Pt and Hg. AB - Two important challenges confronting RNA crystallographers are producing crystals and finding isomorphous heavy-atom derivatives. Non-isomorphism can be addressed by determining the phases using the multiwavelength anomalous dispersion (MAD) method. These phases can be greatly improved by combining phases from MAD experiments done on different heavy-atom derivatives. Heavy-atom derivatives can be created by chemically modifying the RNA through covalent attachment of bromine or mercury to C5 of pyrimidines or [Pt(NH3)3]2+ to N7 of guanine. While phosphorothioates can provide mercury binding sites, disorder can reduce their value for phase determination. The location of these chemical modifications is critical since crystallization of these derivatized RNAs is sensitive to heavy atom induced conformational alterations and crystal packing. PMID- 9399147 TI - Correlation of hydrophobicity and packing in A-DNA oligonucleotides. AB - A-DNA oligomers pack in a slanted fashion with the terminal base pairs abutting into the minor groove of neighboring molecules unlike the other forms of DNA which pack by vertically stacking one over the other into helical columns. To explain the differences in packing we have advanced a hypothesis that the orientation of the sugar-phosphate backbone is different in A-DNA from that in the other forms of DNA, mainly due to the differences in the sugar puckering. PMID- 9399148 TI - The SRY cantilever motif discriminates between sequence- and structure-specific DNA recognition: alanine mutagenesis of an HMG box. AB - The high-mobility-group (HMG) box defines a DNA-bending motif conserved among architectural transcription factors. A "hydrophobic wedge" at the protein surface provides a mechanism of DNA bending: disruption of base stacking by insertion of a sidechain "cantilever." First described in the mammalian testis-determining factor SRY, the cantilever motif consists of adjacent aromatic and nonpolar sidechains at the crux of the HMG box (residues 12 and 13). Here, the role of these side chains in DNA recognition is investigated by alanine mutagenesis. F12A and I13A substitutions in the SRY HMG box each permit native folding and thermal stability (as monitored by circular dichroism and 1H-NMR) but eliminate sequence specific DNA-binding activity (as detected by gel-mobility shift). On binding to the sharp angles of a four-way DNA junction (4WJ), however, the substitutions each promote formation of a high-molecular-weight aggregate, presumably by DNA dependent oligomerization. The substitutions have opposite effects on initial binding to the 4WJ: whereas such binding is attenuated ten-fold by F12A, it is enhanced by I13A. A foreshortened "alanine cantilever", not observed among specific HMG boxes, occurs in a non-specific domain (HMG-1A) and may enhance architecture-selective DNA recognition. PMID- 9399150 TI - Variability analysis of HIV-1 gp120 V3 region: I. Point estimators for the amino acid distribution characteristics. AB - Enumerating procedure for symbol sequences is proposed. Relationship between Hamming distance for symbol sequences and Euclidean distance for corresponding enumerations is established, and more universal Hamming-transformed Euclidean measure is constructed. A distribution function of amino acid substitutions and some of its point estimators (consensus, subconsensus, sample mean, sample central moments and asymmetry coefficient) are introduced. Hamming-transformed Euclidean measures between consensus, subconsensus and sample means for ten HIV-1 taxons of gp120 V3 regions are calculated. It is demonstrated that these taxons have a complicated pattern which is significant for their classification. PMID- 9399149 TI - Structure of an anti-HIV-1 hammerhead ribozyme complex with a 17-mer DNA substrate analog of HIV-1 gag RNA and a mechanism for the cleavage reaction: 750 MHz NMR and computer experiments. AB - The structure of an anti-HIV-1 ribozyme-DNA abortive substrate complex was investigated by 750 MHz NMR and computer modeling experiments. The ribozyme was a chimeric molecule with 30 residues-18 DNA nucleotides, and 12 RNA residues in the conserved core. The DNA substrate analog had 17 residues. The chimeric ribozyme and the DNA substrate formed a shortened ribozyme-abortive substrate complex of 47 nucleotides with two DNA stems (stems I and III) and a loop consisting of the conserved core residues. Circular dichroism spectra showed that the DNA stems assume A-family conformation at the NMR concentration and a temperature of 15 degrees C, contrary to the conventional wisdom that DNA duplexes in aqueous solution populate entirely in the B-form. It is proposed that the A-family RNA residues at the core expand the A-family initiated at the core into the DNA stems because of the large free energy requirement for the formation of A/B junctions. Assignments of the base H8/H6 protons and H1' of the 47 residues were made by a NOESY walk. In addition to the methyl groups of all T's, the imino resonances of stems I and III and AH2's were assigned from appropriate NOESY walks. The extracted NMR data along with available crystallographic data, were used to derive a structural model of the complex. Stems I and III of the final model displayed a remarkable similarity to the A form of DNA; in stem III, a GC base pair was found to be moving into the floor of the minor groove defined by flanking AT pairs; data suggest the formation of a buckled rhombic structure with the adjacent pair; in addition, the base pair at the interface of stem III and the loop region displayed deformed geometry. The loop with the catalytic core, and the immediate region of the stems displayed conformational multiplicity within the NMR time scale. A catalytic mechanism for ribozyme action based on the derived structure, and consistent with biochemical data in the literature, is proposed. The complex between the anti HIV-1 gag ribozyme and its abortive DNA substrate manifests in the detection of a continuous track of A.T base pairs; this suggests that the interaction between the ribozyme and its DNA substrate is stronger than the one observed in the case of the free ribozyme where the bases in stem I and stem III regions interact strongly with the ribozyme core region (Sarma, R. H., et al. FEBS Letters 375, 317-23, 1995). The complex formation provides certain guidelines in the design of suitable therapeutic ribozymes. If the residues in the ribozyme stem regions interact with the conserved core, it may either prevent or interfere with the formation of a catalytically active tertiary structure. PMID- 9399151 TI - Variability analysis of HIV-1 gp120 V3 region: II. Hierarchy of taxons. AB - In the previous work (M. Yu. Shchelkanov, A. N. Yudin, A. V. Antonov, N. S. Starikov, A. A. Vedenov, E. V. Karamov, J. Biomol. Struct. Dyn. 15, 217-229 (1997)) we have introduced the amino acid distribution function within HIV-1 taxons and Hamming-transformed Euclidean measures between their characteristics: consensus, subconsensus and sample mean. In this work the referred characteristics are used for hierarchical classification of amino acid sequences of gp120 V3 region belonging to different HIV-1 taxons. A comparative analysis of the results produced by various classification methods is carried out. Multidimensional scaling of distance matrix for the specified characteristics is used to visualize the pattern of HIV-1 variability. PMID- 9399152 TI - Single-strand-preferring RNases degrade double-stranded RNAs by destabilizing its secondary structure. AB - To establish the mechanism of dsRNA degradation by mammalian single-stranded preferring ribonucleases, and, in particular, the influence of their positively charged non-catalytic amino acid residues, we have studied the kinetic parameters of the depolimerization of single- and double-stranded polyribonucleotides such as poly(U), poly(U).poly(A), poly(C) and poly(C).poly(I) by the action of human seminal RNase, bovine seminal RNase and ox pancreas RNase A. While the activities of these RNases on poly(I).poly(C) were definitely lower than those on poly(C), the activities of human seminal and bovine seminal RNases on poly(U).poly(A) and poly(U) were of the same order of magnitude under physiological salt conditions. The ratio of the RNase A degrading activities towards poly(U) and poly(U).poly(A) at I = 0.16 M is ten times higher than the corresponding ratios determined with bovine seminal and human seminal ribonucleases. The high activities of these two RNases towards poly(U).poly(A) are discussed on the basis of their efficient estabilishing action on this double-helical nucleic acid due to their high affinity for poly(A). The destabilizing action of human seminal RNase and bovine seminal RNase on the poly (U).poly(A) duplex is higher than that measurable with bovine RNase A because of the higher number of positive charges present on those enzyme molecules. This may therefore explain why human seminal and bovine seminal ribonucleases are more efficient than RNase A in the depolymerization of poly(U).poly(A) at physiological ionic strength. PMID- 9399153 TI - Thermodynamic cycle between DNA and RNA constituents for conformation of the sugar ring from nuclear magnetic resonance study. AB - The effect of a structural change of ribose to deoxyribose, by replacement of 2' OH by 2'-H, on the conformational equilibrium of the sugar ring is described in terms of one thermodynamic cycle. The method is based on the observation that conformational correlations of the sugar ring--side chain ensemble in DNA and RNA components show one general pattern, reflecting an intrinsic physical property of this ensemble. The pattern determines a choice of model systems to study. The systems consist of pairs of DNA and RNA components, nucleosides and nucleotides in aqueous solution, where all conformational factors are fully controlled. This approach allowed us to describe the thermodynamic cycle and measure its fundamental parameters, equilibrium constants and free energy differences, delta delta G, from a nuclear magnetic resonance study. The delta delta G values as determined for pairs of ribo- and deoxyribo-nucleosides in classes of syn constrained and anti-preferred models, are comparable and lie in a narrow range, delta delta G = 1.7 +/- 0.1 [kJ/mol]. For pairs of ribo- and deoxyribo nucleotides, the delta delta G values also lie in narrow ranges, delta delta G = 1.7 +/- 0.1 [kJ/mol] for 5'-phosphate nucleotides and delta delta G = 1.9 +/- 0.1 [kJ/mol] for 3'-phosphate nucleotides, i.e. similar to those observed for nucleosides. The measured quantity, delta delta G, is generally observed in a relatively narrow range, delta delta G = 1.75 +/- 0.15 [kJ/mol], irrespective of the class of the model system. This quantity represents a "pure" constant contribution, pe one sugar moiety, as a "driving force" for the N-->S shift in the sugar ring conformational equilibrium, when one compares RNA and DNA. This important thermodynamic quantity, delta delta G, has not hitherto been determined for nucleic acids. Ultimately the delta delta G quantity is revealed in the tendency to adopt S(C2'endo) sugar puckering domain by the majority of DNA structures, whereas RNA generally adopt an N(C3'endo) puckering domain. A possible biological significance of the delta delta G quantity may include evolutionary aspects of nucleic acids. PMID- 9399154 TI - Curvature and sequence analysis of eukaryotic promoters. AB - We have calculated the curvature of 504 eukaryotic promoters predicted by the bent A-tract model of Bolshoy et al. (Proc. Natl. Acad. Sci. USA, 88(6), pp. 2312 16) and the bent non-A-tract models of Calladine et al. (J. Mol. Biol., 201, pp. 127-37) and Satchwell et al. (J. Mol. Biol., 191, pp. 659-75) and found in each case a correlation between TBP binding sites and DNA curvature. Characterizing the TBP binding sites revealed that in addition to the classical TATA box (TATAAA) five more elements occur significantly often in the promoters, nearly all of them being one point mutations of the classical TATA box element. Separate curvature calculations for promoters with canonical and non-canonical TATA boxes have shown that in both cases the strong curvature of the helix axes in the domain of the binding sites is maintained (classical TBP binding sites: + 64 135%, non-classical TBP binding sites: + 27-49%). These results support the proposition that beside DNA flexibility and DNA-protein interactions intrinsic curvature of DNA is one further important criterion for the recognition of different DNA elements by TBP. PMID- 9399155 TI - An improved method for the rapid assessment of DNA bending by small molecules. AB - Assessing the effects of non-covalently bound chemicals on DNA structure is particularly challenging. Traditional methods require the use of cumbersome electrophoretic techniques or that the compounds bind DNA with an extremely high affinity. Here we demonstrate that, by extending the use of the exonuclease Bal 31, we can rely on a standard cyclization assay technique and one dimensional gel electrophoresis to identify and quantitate chemical induced DNA bending. An important application of this method is to the study of small molecules that bind to DNA non-covalently and we illustrate the method with the antitumor antibiotic calicheamicin. Our results suggest that the distribution of circles resulting from the polymerization of a 21 bp DNA construct reflects the kinetics of the competing cyclization and dimerization reactions and provides a method for rapidly screening compounds for DNA bending. PMID- 9399156 TI - Structure of poly d(AI).poly d(CT) in two different packing arrangements. AB - X-ray diffraction analysis of poly d(AI).poly d(CT) in oriented and polycrystalline fibers has revealed the DNA structure to be a 10-fold, right handed, antiparallel, Watson-Crick base paired double helix in two distinct packing arrangements corresponding to one and two helices, respectively, in the unit cell. The helix pitch is 32.1 A and 32.4 A in the two cases, almost 1.5 A shorter than in classical B-DNA. The resulting B'-DNA geometry, described in terms of a tetranucleotide repeat which is conformationally similar to B-DNA, has its minor groove closely shut and major groove correspondingly widened, thus striking a sharp morphological contrast to B-DNA. According to difference electron density maps, a spine of hydration along the minor groove connects both strands and provides structural stability; ordered sodium ions and water molecules are actively involved in bridging the phosphate groups of neighboring helices. The crystallographic R-values for these two allomorphs are 0.26 and 0.20, respectively, for data up to 3.0 A resolution. PMID- 9399158 TI - Dynamic bis-intercalation of a homodimeric thiazole orange dye in DNA: evidence of intercalator creeping. AB - We have used one and two dimensional exchange 1H NMR spectroscopy to characterize the dynamics of the binding of a homodimeric thiazole orange dye, 1,1'-(4,4,8,8 tetramethyl-4,8-diaza-undecamethylene)-bis- 4-(3-methyl-2,3-dihydro-(benzo-1,3 thiazole)-2-methylidene)-quinol inium tetraiodide (TOTO), to double stranded DNA (dsDNA). The double stranded oligonucleotides used were d-(CGCTAGCG)2 (1) and d(CGCTAGCTAGCG)2 (2). TOTO binds preferentially to the (5'-CTAG-3')2 sites and forms mixtures of 1:1 and 1:2 dsDNA-TOTO complexes with 2 in ratios dependent on the relative amount of TOTO and the oligonucleotide in the sample. The dynamic exchange between preferential binding sites in the case of a 2:1 1-TOTO mixture is an intermolecular exchange process between two binding sites on different oligonucleotides. In the case of the 1:1 2-TOTO complex an intramolecular exchange process occur between two different binding sites on the same strand. Both processes were studied. The results demonstrate the ability of TOTO to migrate along a dsDNA strand in an intramolecular exchange process. The migration process ("creeping") along the DNA strand is 6 times faster than the rate of intermolecular exchange between sites in two different oligonucleotides. PMID- 9399157 TI - Structural studies by high-field NMR spectroscopy of a binary-addressed complementary oligonucleotide system juxtaposing pyrene and perfluoro-azide units. AB - Recently, a new approach has been proposed to improve the site-specificity and efficiency of the modification of nucleic acid target sequences, the binary system of complementary-addressing nucleic acid sequences. The binary system comprises two oligonucleotides, one modified with a photosensitizing group and the other with a photoreactive group. The sites of chemical modification are arranged to bring the two chemical functions close enough together in space to allow efficient energy transfer from the photo-excited photosensitizer to an arylazide moiety which expels N2 to form a nitrene which subsequently covalently labels the target nucleic acid. Structural analysis performed by high-resolution 2D NMR spectroscopy (400 MHz and 600 MHz) are reported for the model binary system 1:2:3, where 1 is the target 12-mer pdGTATCAGTTTCT, 2 is a photoactivatable fluoroazide derivative dAGAAACp-L-Az and 3 is the photosensitizer derivative Pyr-pdTGATAC (here: Az is the p-azidotetrafluorobenzyl group, Pyr the pyrenyl-1-methylamino group, L a linker group). The assignment of oligonucleotide and modifying group protons was performed using 1H COSY, TOCSY and NOESY experiments. Comprehensive analysis of 1H NOESY spectra of 1:2:3 showed that terminal fragments of the complex [5'p-1T-2G-3A-4T-], [-21A-22T-23A-24C], [ 8T-9T-10T-11C-12T] and [13A-14G-15A-15A-17A-18C-] gave a continuous set of intra- and inter-nucleotide interactions, typical of regular double-stranded B-DNA. In contrast, the central region of the complex composed of 5C, 6A, 7G, 19T and 20G nucleotide residues, nearest the Pyr and Az groups, was found to be distorted. Thus some signals from aromatic and/or sugar-ring protons of the above nucleotide residues were extremely broadened or almost absent. Moreover, some intra- and/or inter-nucleotide interactions, typical of the regular DNA duplex, were not detected for the [-5C-6A-7G-] and [-19T-20G-] regions of the tandem system. Instead of that, some cross-peaks of low-intensity between the H2 proton of the Pyr group and 7G(H1'), 7G(H2'/H2"), 7G(H3'), 4T(H2"), 4T(H4') and 4T(H5'/H5") were observed. Additional 1H -1H NOE-interactions between methylene protons of the linker group L and some sugar ring protons of 18C nucleotide residue were detected. A preliminary structural model, constructed using proton-proton distances between Pyr and the DNA and Az-L and DNA obtained from a 1H NOESY experiment at 300 ms mixing time as constraints for the refinement of the structure, displayed significant distortion from B-DNA of the double-stranded helix in the middle of the complex, (-5C-6A-7G, -18C-19T-20G-). The Pyr group was located in what remains of the minor groove near 4T, 5C, 6A and 7G and the centroid of the azide ring less than 9A degrees from the centroid of the ring system of Pyr group. PMID- 9399159 TI - Sequence-independent recombination triple helices: a molecular dynamics study. AB - Recent experimental studies have shown that the Rec-A mediated homologous recombination reaction involves a triple helical intermediate, in which the third strand base forms hydrogen bonds with both the bases in the major groove of the Watson-Crick duplex. Such 'mixed' hydrogen bonds allow formation of sequence independent triplexes. DNA triple helices involving 'mixed' hydrogen bonds have been studied, using model building, molecular mechanics (MM) and molecular dynamics (MD). Models were built for a triplex comprising all four possible triplets viz., G.C*C, C.G*G, A.T*T and T.A*A. To check the stability of all the 'mixed' hydrogen bonds in such triplexes and the conformational preferences of such triplex structures, MD studies were carried out starting from two structures with 30 degrees and 36 degrees twist between the basepairs. It was observed that though the two triplexes converged towards a similar structure, the various hydrogen bonds between the WC duplex and the third strand showed differential stabilities. An MD simulation with restrained hydrogen bonds showed that the resulting structure was stable and remained close to the starting structure. These studies help us in defining stable hydrogen bond geometries involving the third strand and the WC duplex. It was observed that in the C.G*G triplets the N7 atom of the second strand is always involved in hydrogen bonding. In the G.C*C triplets, either N3 or O2 in the third strand cytosine can interchangeably act as a hydrogen bond acceptor. PMID- 9399161 TI - Search for rigid sub domains in DNA from molecular dynamics simulations. AB - A strategy is presented for searching which atoms can be regrouped within rigid sub-units during the time course of Molecular Dynamics simulations of biopolymers. The root mean square fluctuations of the interatomic distances are used as a criterion. The number of rigid sub-units which are found depends on the tolerance rc for the definition of a rigid body, i.e. until which value the fluctuations can be neglected. The method is applied to two self-complementary oligonucleotides belonging to the B-form family which give identical results. With rc = 0.027 nm each nucleotide may be described as 3 rigid sub-units: the sugar ring, the base and the backbone (PO4 + C5' atoms). With rc = 0.01 nm, the same sub-units are obtained except that C5' can no more be regrouped with the PO4 atoms. It is shown that the variation of the coulombic potential owing to the deformation of the sub-units during the time course of the simulation is on the same order of magnitude as the inaccuracy due to the choice of the force field parameters. PMID- 9399160 TI - Protein-nucleic acid recognition: simulation of base and "model" amino acids complexes in DMSO by the Monte Carlo method. AB - A computer simulation of guanine (G), cytosine (C), the G-C base pair, protonated C (CH+), acetic acid in neutral (AcOH) and deprotonated (AcO-) forms, G-AcO-, C AcOH, and CH(+)-AcO- complexes, solvated in DMSO was carried out by the Monte Carlo method. It is shown that the G-C base pair formation in DMSO is energetically favorable. The G-AcO- complex formation is comparable with the formation of G-C base pair in energetically favorability. In this case the acetate anion can replace C in the G-C base pair. The formation of the C-AcOH complex is much less favorable than the formation of the G-C pair. However proton transfer from AcOH to C leads to the formation of the CH(+)-AcO- complex, which is the most favorable of all complexes studied. Here the acetic acid can replace G in a G-C base pair. The formation of G-AcO- and CH(+)-AcO- specific complexes detected in DMSO with the help of experiment and theory is a competitive process with respect to the formation of G-C base pairs, and can be considered the primary step in the real mechanism of protein-nucleic acid recognition. PMID- 9399162 TI - Cooperative interactions of the oligodeoxyribonucleotides on the complementary template. The influence of chemical groups and mismatched nucleotides at the 5'- and 3'-ends of oligonucleotides on the parameters of cooperativity. AB - Parameters of cooperative interactions of two or three oligodeoxyribonucleotides or their derivatives bound with the adjacent sites of the complementary template were measured using method of "complementary addressed modification titration" (CAMT). Complementary template (target) were modified with the reactive oligonucleotide derivatives (reagents) bearing covalently attached alkylating 4 [N-(2-chloroethyl)-N-methylamino]benzylamino- group (C1RCH2NH)- at 5'-terminal phosphate. The targets had only one binding site for the reagent and either no (T10), or one (T'22 and T22) or two sites (T26) for the oligonucleotides (effectors) cooperatively bound with the adjacent sites on the template. Both unmodified oligonucleotides E1, E2 and their derivatives E1Phn, E2Phn bearing N (2-hydroxyethyl)-phenazinium residues Phn- both at 5'- and 3'-ends covalently linked via ethylenediamine linker were used as effectors. Effectors E1 and E2 (E1Phn and E2Phn) bind, respectively, upstream or downstream from the reagent. Hexameric (X6) or octameric (X8 or X8m) reagents were used for the target modification. The reagent X8m formed one TT-mismatch with the target at the end opposite to location of the reactive moiety. The cooperativity parameter values characterizing the mutual interactions between the reagents X6, X8, X8m and effectors E1, E2, E1Phn, E2Phn have been found as the ratio of the association constants of the reagents in the presence of effectors. The association constants were calculated from the dependencies of the target modification extent on initial concentrations of the reagents. The use of T26 existing both in linear and hairpin conformations permitted us to estimate additionally the role of indirect cooperativity originating from the induction of the target conformational change by the effectors. The following conclusions were done from the quantitative results. The efficiency of direct cooperativity is independent on the length of oligonucleotide for the same nature of the contact. The cooperativity parameter increases by factor about 3 in the presence of Phn-group covalently attached to oligonucleotides and located at the junctions. The presence of either alkylating group C1RCH2NH- or TT-mismatch at the junctions eliminates cooperative interaction between the bases. In the same time sufficiently effective cooperative interaction takes place in the case of simultaneous presence of both Phn- and either C1RCH2NH- group or TT-mismatch at the junction. PMID- 9399163 TI - Dielectric relaxation, molecular motion and interprotein interactions in myoglobin solution. AB - The results of the investigation of protein molecule dynamic in solution by Time Domain Dielectric Spectroscopy are presented. The horse myoglobin solutions in wide range of concentration from 0.6% to 54% at 20 degrees C have been investigated. The result of analysis produced in the term of dipole correlation function has shown that the obtained correlation function of macromolecule motion may be presented as sum of three components corresponding to three kinds of protein motions: anisotropic intramolecular motion, anisotropic Brownian tumbling and isotropic slow motion. We suppose that the cause of protein tumbling anisotropy and the possibility to keep slow motion is the interprotein electrostatic interactions. The characteristic time of slow motion depends on the concentration of protein and perhaps is controlled by translational diffusion. The dipole moment of myoglobin calculated by the Onsager-Oncley equation is 200D for solutions less than 10% protein concentration. It is in a good agreement with the theoretical value. PMID- 9399164 TI - Modeling of in vivo proteolytic degradation of hemoglobin. AB - Based on the amino acid sequences of endogenous peptides and X-ray spatial structure, mechanism of the in vivo proteolly degradation of bovine hemoglobin was analysed. The degradation was shown to be a multi-stage process. Its first stage is determined by the spatial organization of the native protein substrate, and the next stages-by the distribution of the electrostatic field potential of the protein fragments formed at the earlier stage. PMID- 9399165 TI - Aromatic-aromatic ring interaction revisited with model compounds of Wilcox. AB - Aromatic-aromatic nonbonded interactions have been reexamined using model compounds of Wilcox and collaborators (J. Am. Chem. Soc. 1994, 116, 4497). It was found that at low temperatures down to 210 degrees K, the population of the folded conformers (A) is higher than that of the unfolded conformers (B), suggesting that edge-to-face aromatic-aromatic ring interactions are in effect. However, the free energy difference between the two types of conformers did not vary linearly with temperature, which is against what we expected from the thermodynamic relationship of delta G = delta H-T delta S. This suggests that in the presence of solvent molecules a free energy cancellation effect operates between the two conformers. Although A has a free energy gain of only approximately 0.5 kcal/mol over B in organic solvents, as obtained by subtracting the solvent-induced unfolding effect, it could still be a significant energy with respect to conformational preference. PMID- 9399166 TI - Current status of computed radiography in emergency departments. AB - This study reports the findings of a computed radiography (CR) imaging experience questionnaire sent to 35 emergency departments (ED) in North America. A total of 25 responses to the questionnaire were received corresponding to a return rate of 71%. The median daily workload was 71 patient examinations and the average number of films per patient examination for the 21 facilities was 3.0 +/- 0.8. A total of 91% of respondents printed to film all ED trauma images obtained with CR with only one ED claiming to be filmless. CR in the ED was easy to use and had significant benefits of reducing examination repeat rates, permitting a prompt availability of radiographic images, improving image quality, providing improved operational efficiency, and eliminating lost films. Major limitations of CR were deemed to be limited viewing stations, CR costs, and inefficient patient ID entry. Radiology departments were very happy with the introduction of CR into the ED setting with approximately half being highly satisfied and half somewhat satisfied. The degree of satisfaction by ED personnel was similar with about half being highly satisfied, 40% somewhat satisfied, and the remainder neutral. The fact that not a single respondent was in any way dissatisfied shows that CR can play a useful role in the ED setting. PMID- 9399167 TI - Eye-tracking device comparisons of three methods of magnetic resonance image series displays. AB - This study evaluated the effectiveness of three kinds of display methods for magnetic resonance (MR) image interpretation using an eye-tracking device. Seven radiologists interpreted head MR studies by using a single monitor (17-inch, 1,024 X 1,280 bit) in the 4 images/screen display format. Three paging modes were compared: (A) rapid paging only, (B) multiple image series display at the same slice position with consecutive rapid paging, and (C) simultaneous display of multiple series with each image series being browsed independently. Using an eye mark camera, the radiologist's point of fixation and the duration of fixation were recorded during actual image interpretation. In mode A, the duration of fixation was short, and the points of fixation were distributed randomly over the visual field. In mode B, the points of fixation were clustered chiefly on a specific image series. In mode C, the points of fixation were not clustered on a specified series, but the duration of viewing the T2 series was relatively long. The total tracing area in mode B and C was smaller than that in mode A. Multiple series display, in which selected key series of slices could be viewed effectively, was found to be suitable for MR image interpretation. PMID- 9399168 TI - Receiver operating characteristic study of image preprocessing for teleradiology and digital workstations. AB - The purpose of this study was to evaluate whether digitized analog images displayed on a digital workstation can be improved by using a preprocessing algorithm, and if so, whether the quality of the resulting images can reach that of the original films. The material contained 120 difficult cases (about 50% with selected pathology). Four radiologists each evaluated half of the randomly ordered cases with the digital workstation and half of the cases with the original radiographs. The data were compared with a previous similar study, where the workstation had no option for preprocessed images. Preprocessed digital images were clearly superior to digital images without preprocessing, although for those of the highest diagnostic difficulty they were inferior to the original films. The preprocessing algorithm has improved the diagnostic quality of the digital workstation. There is room yet for improvement compared to plain films, although the current setup may be sufficient in some settings. PMID- 9399169 TI - A brief review of human perception factors in digital displays for picture archiving and communications systems. AB - The purpose of this review is to further inform radiologists, physicists, technologists, and engineers working with digital image display devices of issues related to human perception. This article will briefly review the effects of several factors in human perception that are specifically relevant to a digital display environment. These factors include the following: the spatial and contrast resolution of the display device; background luminance level and luminance range of the display system; brightness uniformity; extraneous light in the reading room; displayed field size; viewing distance; image motion and monitor flickering; signal to noise ratio of the displayed image; magnification functions; and the user interface. After reviewing the perception study results, a checklist of desirable features and quality assurance issues for a digital display workstation are presented as an appendix. PMID- 9399170 TI - The effect of lossy discrete cosine transform compression on subtle bone fractures. AB - Extensive research efforts have been devoted to the feasibility of picture archiving and communication systems (PACS) in recent years. The advantages of PACS are numerous but mainly include reduced cost and improvement in the operational efficiency of a PACS-based radiology department. In digital radiography, images are viewed either in hard-copy or soft-copy format. Usually, these images are subsequently compressed and archived for future evaluation. There are various methods used in image compression. In this study, computed radiography images showing subtle pediatric bone fractures were compressed with the lossy method of image compression after they had been initially evaluated on workstation monitors. These studies were subsequently evaluated by observers, who were unaware of the interpretations of these images before compression, to determine if they could detect similar abnormalities. Our conclusion is that there is no difference in the interpretation of soft-copy computed radiographic images before or after lossy 10:1 compression in studies of subtle pediatric bone fractures. This is a US government work. There are no restrictions on its use. PMID- 9399172 TI - The Society for Computer Applications in Radiology. PMID- 9399171 TI - The effect of intensity windowing on the detection of simulated masses embedded in dense portions of digitized mammograms in a laboratory setting. AB - The purpose of this study was to determine whether intensity windowing (IW) improves detection of simulated masses in dense mammograms. Simulated masses were embedded in dense mammograms digitized at 50 microns/pixel, 12 bits deep. Images were printed with no windowing applied and with nine window width and level combinations applied. A simulated mass was embedded in a realistic background of dense breast tissue, with the position of the mass (against the background) varied. The key variables involved in each trial included the position of the mass, the contrast levels and the IW setting applied to the image. Combining the 10 image processing conditions, 4 contrast levels, and 4 quadrant positions gave 160 combinations. The trials were constructed by pairing 160 combinations of key variables with 160 backgrounds. The entire experiment consisted of 800 trials. Twenty observers were asked to detect the quadrant of the image into which the mass was located. There was a statistically significant improvement in detection performance for masses when the window width was set at 1024 with a level of 3328. IW should be tested in the clinic to determine whether mass detection performance in real mammograms is improved. PMID- 9399173 TI - And thus, the hand revealed its beauty. Moving toward aesthetics. PMID- 9399174 TI - The architects of the future. PMID- 9399175 TI - Fractures of the distal radius: what are the expectations of therapy? A two-year retrospective study. AB - This retrospective study addressed two basic questions important to hand therapists: what are the expectations of therapy vis-a-vis range of motion and strength among patients with distal radius fractures, and do patients treated by different therapists within a center experience substantially similar or dissimilar outcomes? Differences between patient groups were tested using ANOVA, and changes in performance were tested using the Student's t-test; significant differences between patient groups treated by different therapists were lacking and patients demonstrated significant changes in active motion and strength. Data were used to derive regression equations that yielded good estimates of active motion at discharge from therapy. Patients were treated an average of ten times; at discharge, they demonstrated active wrist and forearm motion compatible with the performance of activities of daily living. PMID- 9399176 TI - Pillar pain as a postoperative complication of carpal tunnel release: a review of the literature. AB - Carpal Tunnel Syndrome (CTS) has been referred to as the most common peripheral entrapment neuropathy. As Mirza and colleagues note, its incidence continues to increase. Einhorn and Leddy cite Palmer's estimated incidence of 1% in the general population and 5% or more of workers in certain industries which require repetitive use of the hands and wrists. Conservative treatment of CTS includes splinting and modification of activities. However, surgical release of the transverse carpal ligament or the flexor retinaculum is an extremely common procedure. The open surgical technique has been used since 1924 and is still considered by many to be the gold standard. In 1989 Oksuto introduced the endoscopic carpal tunnel release (ECTR) with the rationale of potentially decreasing the prevalence of complications. In the ensuing years, endoscopic results have generated a tremendous amount of study and controversy. Berger reported that many "passionate arguments both for and against the use of ECTR" exist. This paper briefly reviews the literature generated by this debate, focusing on one potential postoperative complication: pillar pain. Various definitions of pillar pain are noted, and suggested etiologies are grouped into four categories. This is followed by a brief discussion of the treatment approaches and issues. PMID- 9399177 TI - The effect of use of a wrist orthosis during functional activities on surface electromyography of the wrist extensors in normal subjects. AB - Wrist orthoses are advocated for patients with lateral epicondylitis on the assumption that use of the orthosis decreases muscle activity of the wrist extensors during activities. The purpose of this study was to compare the amount of electrical activity, root mean square (RMS) calculated from surface EMG recorded over the wrist extensors, during activities when applying four conditions of wrist orthoses: dorsal; volar; semicircular; and no orthosis. Thirteen normal subjects (mean age 27.7 years) performed three lifting and two gripping tasks, repeated on three consecutive days under four orthotic conditions. Measured were RMS and maximum voluntary grip strength. Repeated measures ANOVA's indicated a significant decrease of RMS using the semicircular design during lifting (p < .0005). Grip strength decreased significantly using all three orthotic designs, but RMS recorded during gripping did not. It was concluded that application of a wrist orthosis reduces electrical activity of the wrist extensors less than anticipated and only during lifting. PMID- 9399178 TI - Reliability of isometric wrist extension torque using the LIDO WorkSET for late follow-up of postoperative wrist patients. AB - The objectives of this study were to determine interobserver, intraobserver, and overall reliability of isometric wrist extension torque using the LIDO WorkSET and then to estimate the minimal level of detectable change using the standard error of measurement. A generalizability study was conducted on 18 postoperative patients. Primary outcome was mean torque of three trials. Variance components were used to calculate generalizability coefficients for intraobserver (G = 0.96), interobserver (G = 0.94), and overall reliability (G = 0.92). When the same therapist is evaluating a patient on different days, a change of more than 16 inch-lb is needed to be 90% confident that true change has occurred. A greater value for change (23 inch-lb) is required when different therapists evaluate a patient on different days. The LIDO WorkSET measures wrist torque in a reproducible manner in an applied setting. The testing protocol is sensitive to differences in wrist torque between individuals and tolerated in the late postoperative period. PMID- 9399179 TI - The force/time relationship of clinically used sensory testing instruments. AB - The stimuli of commonly used sensibility measurement instruments tested in this study demonstrate unequivocally that "hand-held instruments" produce variations in application force from one stimulation to another, one instrument to another, and from one examiner to another. These application force variations cannot be compensated for by care in technique and need to be controlled for measurement reliability. Only the Semmes-Weinstein monofilaments provide some control of force during application and can be considered force controlled if calibrated and applied correctly. The monofilaments, too, become less controllable if applied too quickly and bounced against the skin. Two-point discrimination instruments, in particular, lack control of application force, with the force of one point significantly different from two points. A difference in applied force makes it possible for a patient to solve the two-point discrimination test by discerning the difference between heavier and lighter forces, rather than one or two-point recognition. Tuning fork instruments for vibration testing have even larger variations in application force amplitude rendering their stimulus highly uncontrolled and masking the actual vibration of the tuning fork. Spectral analysis of the force frequency signal produced by hand held sensibility measurement instruments shows they all produce both high and low frequency signals sufficient in strength to stimulate both slowly adapting and quickly adapting end organs, and are not capable of stimulating one particular group. These dynamic properties of testing stimuli explain why our tests are not as repeatable and sensitive as desired. The understanding of these dynamic properties in sensibility measurement is a key for improved instruments and more repeatable findings in clinical testing. PMID- 9399180 TI - The clamshell digit splint with Otoform lining. PMID- 9399181 TI - A static progressive splint for Dupuytren's release. PMID- 9399182 TI - Defining handedness: a critical variable. PMID- 9399183 TI - A review of esthetic alternatives for the restoration of anterior teeth. AB - PURPOSE: This article describes different options for the esthetic treatment of anterior teeth, starting with minimally invasive procedures, such as facial surface bleaching and bonding with composites. METHODS: The importance of metal ceramic restorations, porcelain shoulder techniques, and metal free ceramics are also emphasized. The options are carefully demonstrated to identify advantages and limitations of each technique. PMID- 9399184 TI - Temperature rise in pulpal chamber during fabrication of provisional resinous crowns. AB - STATEMENT OF PROBLEM: The heat generated during the exothermic polymerization reaction of autopolymerizing resinous materials and the heat generated by ultraviolet lamps during irradiation of photopolymerizing resinous materials could cause pulpal damage when a direct technique is used to fabricate provisional restorations. This could occur if temperature elevations overcome the physiological heat dissipating mechanisms of the dental-periodontal system. PURPOSE: This in vitro study compared the rise in temperatures in the pulpal chamber during fabrication of provisional complete veneer crowns by direct method with different autopolymerizing and photopolymerizing resins. The effect of curing resinous crowns in different matrices, such as a polyvinyl siloxane impression and a vaccuum-formed polypropylene sheet, was also evaluated. RESULTS: The results demonstrated that the amount of heat generated during resin polymerization and transmitted to the pulpal chamber could be damaging to pulpal tissues including odontoblasts. When curing of provisional resinous crowns was performed in the polyvinyl siloxane impression, significantly lower temperatures were recorded compared with curing in the vacuum-formed polypropylene sheet. CONCLUSIONS: To prevent pulpal damage, effective cooling procedures are strongly recommended when directly fabricating resinous provisional crowns. PMID- 9399186 TI - Effect of different surface textures on retentive strength of tapered posts. AB - STATEMENT OF PROBLEM: Tapered posts allow for the preservation of tooth substance in the fragile apical area and are advantageous in clinical situations where they conform to the root and canal configuration of endodontically treated teeth. However, their lower retention compared with passive parallel-sided or active threaded posts is a disadvantage. PURPOSE: This study determined the retentive strength of tapered titanium posts with different surface textures and examined the effect of roughening dentinal walls of the prepared post space. MATERIAL AND METHODS: Posts with four surface configurations (smooth, with and without grooves, and sandblasted, with and without grooves) were examined when cemented in extracted anterior teeth. RESULTS: The smooth post showed the lowest retentive strength. Sandblasting the smooth post more than doubled its retentive strength. The retentive strength of both smooth and sandblasted posts could be further increased by the addition of circumferential grooves. Roughening the dentinal walls of the prepared post space increased the retentive strength of sandblasted posts with and without grooves even more. CONCLUSIONS: These findings indicated that, when a tapered post is used, roughening the dentin canal wall, as well as sandblasting and grooving the post, can provide statistically significant additional resistance to dislodgment. PMID- 9399185 TI - Fracture strengths of provisional restorations reinforced with plasma-treated woven polyethylene fiber. AB - STATEMENT OF PROBLEM: Fracture strength of interim fixed partial prosthesis is of great concern, especially in long-span restorations or areas of heavy occlusal stress. PURPOSE: Effects of a plasma-treated woven polyethylene fiber (Ribbond) on the fracture strength of polymethyl methacrylate (Coldpac) and a resin-based two-phase curing provisional restorative material (Provipont DC) were evaluated. MATERIAL AND METHODS: A polyvinyl siloxane template was used to fabricate three unit posterior provisional prostheses on a stainless steel die with two abutments 22 mm apart. The reinforced groups were fabricated by affixing 3 mm wide pieces of fiber treated with methyl methacrylate monomer or polyisocyanate (activator part of Provipont DC) on the occlusal surfaces of abutments. The interim materials were mixed, according to the manufacturers' specifications, and placed in the template. The template was pressed on the die and held secure until complete setting of the material occurred by light curing (Provipont DC) or autopolymerization (PMMA). The specimens were divided into 4 groups of 10 each (A, reinforced Provipont DC; B, unreinforced Provipont DC; C, reinforced PMMA; and D, unreinforced PMMA). A central compressive load force was exerted on the specimen to determine the fracture load of the restorations. RESULTS: The data revealed mean fracture loads of A, 65.59 +/- 11.27 kg; B, 46.59 +/- 14.84 kg; C, 53.46 +/- 7.76 kg; and D, 49.86 +/- 14.44 kg. CONCLUSION: Plasma-treated polyethylene reinforced PMMA restorations showed no significant increase in fracture loads when compared with unreinforced restorations (p > 0.10), whereas reinforced resin-based restorations revealed significantly higher fracture loads (p < 0.01) than the unreinforced resin-based and PMMA provisional restorations. PMID- 9399187 TI - Cyclic fatigue testing of five endodontic post designs supported by four core materials. AB - PURPOSE: This pilot study examined the cyclic fatigue of five endodontic post systems (AccessPost, Flexi-Flange, Flexi-Post, ParaPost, and Vlock) with four core materials (Tytin silver amalgam, Ti-Core, Ketac-Silver and G-C Miracle Mix). MATERIAL AND METHODS: In vitro cyclic fatigue was performed with a machine designed to simulate masticatory fatigue forces. An instantaneous force of 22.2 N (5 pounds) was applied to each post and core combination for a test configuration of 4,000,000 repetitions, or until failure occurred. The type of failure and number of repetitions at failure was recorded for each sample tested. Two-way analysis of variance (ANOVA) was used to compare groups. RESULTS: All posts/core samples with Ti-Core composite and Tytin silver amalgam completed the test with no failures. All posts/core samples with Ketac-Silver material failed before the 4,000,000 test cycle configuration and all failures were core failures. All posts/core samples with G-C Miracle Mix material failed in a similar manner. Newman-Keuls multiple comparison test illustrated that, with this simulated fatigue test, Ti-Core material and Tytin silver amalgam were superior to both G-C Miracle Mix and Ketac-Silver materials. PMID- 9399188 TI - Clinical performance of resin-bonded fixed partial dentures and extracoronal attachments for removable prostheses. AB - STATEMENT OF PROBLEM: It is important to evaluate the long-term clinical performance of resin-bonded fixed partial dentures and extracoronal attachments for removable prostheses. PURPOSE: A prospective, long-term clinical study was conducted to evaluate the success of resin-bonded fixed partial dentures since 1985 and of resin-bonded extracoronal attachments from 1987. METHODS: Until 1993, a total of 130 resin-bonded fixed partial dentures had been seated in 101 patients, as well as 12 removable partial dentures (RPDs) with 24 extracoronal retainers in 10 patients. The clinical treatment protocol and the laboratory procedures were standardized. By the end of 1993, it was possible to reexamine 98 patients with a total of 127 resin-bonded fixed partial dentures and all 10 patients with removable partial dentures. The average time in function for the resin-bonded fixed partial dentures at the time of examination was 3.4 years and 2.3 years for the removable restorations. RESULTS: During the period of observation, one retainer failed on six of the resin-bonded fixed partial dentures, which represents a failure rate of 4.7%. Debonding of extracoronal attachments was recorded for 8.3% of the total number of retainers. CONCLUSION: The resin-bonded fixed partial denture technique can be considered to be a clinically reliable method of treatment, and permits the expansion of indications beyond a classical three-unit resin-bonded fixed partial denture. Long-term clinical success of removable partial dentures with resin-bonded extracoronal retainers warrants additional clinical studies. PMID- 9399189 TI - "Prosthetic condition" and patients' judgment of complete dentures. AB - PURPOSE: This study introduces the concept "prosthetic condition", which combines the quality of complete dentures and residual alveolar ridges. MATERIAL AND METHODS: A pilot study was performed to select quality criteria with an acceptable interobserver agreement. With these criteria, a clinical examination was performed to assess the quality of the existing complete dentures and the residual alveolar ridges of 397 complete denture wearers. During clinical examination, the interobserver agreement of the selected criteria was retested. The "prosthetic condition" was assessed by combining the scores for denture quality and quality of the residual alveolar ridges. Subsequently, participants' satisfaction with and complaints about their dentures were scored according to their answers to specific questions. RESULTS: Logistic regression analysis demonstrated that no variable of the "prosthetic condition" proved to explain the denture satisfaction. Some variables of the "prosthetic condition" had a significant but not relevant correlation with some denture complaints. CONCLUSIONS: More research is necessary to substantiate the concept "prosthetic condition" as an acceptable measure of professionally quality assessment of dentures and denture-bearing surfaces. However, in determining the treatment need of community-dwelling groups, this concept seems a more realistic measure than denture quality only. PMID- 9399191 TI - AllCeram crowns for single replacement implant abutments. AB - Clinicians who are comfortable with traditional porcelain fused to metal restorations may find the thickness of veneering porcelain addition to the CeraOne single tooth ceramic cap disconcerting. When using a premanufactured ceramic cap to fit the space of the final restoration, substantial amounts of "unsupported" veneer porcelain may be required to achieve tooth contact to adjacent or opposing dentition. A potential problem of weak, unsupported veneer porcelain has been addressed by a modification of an existing manufacturing process. By using CAD/CAM technology, a custom-designed Procera AllCeram coping can be created for the implant abutment that eliminates any concerns regarding the resultant design of the underlying ceramic cap substructure. PMID- 9399190 TI - The effect of diet on the bearing mucosa during adjustment to new complete dentures: a pilot study. AB - PURPOSE: This pilot study investigated the reliability of the conventional prosthodontic wisdom that a modified diet ameliorates soreness during the adjustment to new complete dentures. Tissue ulceration of the bearing mucosa served as the indicator of patient soreness as a function of diet. MATERIAL AND METHODS: Fourteen men were randomly assigned to two equal treatment groups. One group consumed a consistency-gradated diet and the other (control) ate as they wished. New complete dentures were fabricated for both groups by the same provider, technician, materials, and method; the study was double-blinded. Tissue ulceration was assessed and totaled for the 10 days after denture placement, constituting a soreness score. RESULTS: Data analysis identified a significant difference in soreness scores between dietary groups (p < 0.05). Wearing time had a significant inverse relation to soreness (p < 0.05) in this study. A host of potential explanatory variables of clinical interest failed to relate significantly to the outcome of soreness. CONCLUSIONS: The results of this study indicated that a consistency-gradated diet had a significant effect in diminishing tissue ulceration during the immediate postplacement period for this group of men. Potentially, these findings could contribute to enhanced quality of care and to the more efficient allocation of provider-based resources. PMID- 9399192 TI - A longitudinal clinical assessment of spark erosion technology in implant retained overdenture prostheses: a preliminary report. AB - STATEMENT OF PROBLEM: As adapted for the dental profession, spark erosion technology permits precise machining of retentive metal overdenture frameworks for use in implant prosthetics. PURPOSE: The resultant prostheses are retentive and provide a number of benefits offered by both conventional overdenture and fixed prosthetic designs. MATERIAL AND METHODS: Preliminary data collected from an ongoing 5-year clinical trial were reviewed to qualitatively assess the clinical results obtained from 25 spark eroded implant-retained overdenture prostheses placed in 24 subjects. RESULTS: Throughout an evaluation period of 13.33 months (range 4 to 19 months), subject responses measured by questionnaire were uniformly good. Few complications were encountered and were limited to resin denture base/tooth fractures or retentive component failures that were easily repaired. CONCLUSION: Overdenture prostheses retained by spark eroded milled frameworks offer an acceptable treatment alternative for patients undergoing dental implant therapy. PMID- 9399193 TI - Effects of implant anchorage on midface prostheses. AB - STATEMENT OF PROBLEM: Acquired midface defects may produce functional and psychologic impairments that adversely effects a patient's quality of life. Conventional prostheses may lack adequate retention and stability, diminishing the patient's confidence that the prosthesis will remain in place during routine activities. PURPOSE: The experience with and patient response to endosseous implants in prosthetic restoration of midface defects is presented in this study. MATERIAL AND METHODS: Five patients in age from 36 to 88 years were treated with 19 titanium endosseous root-form implants to provide retention and stability for prostheses. Patients responded to a questionnaire rating overall use, effectiveness, and satisfaction of their prosthesis, before and after the use of implants. RESULTS: All 19 implants were judged to be osseointegrated at abutment connection. Of the 17 implants used prosthetically, 14 (82%) remained osseointegrated and 3 (18%) failed. Analysis of the questionnaire tends to indicate an improvement of the quality of life for the patients with an implant retained prosthesis. PMID- 9399194 TI - Dynamic mechanical thermal analysis of maxillofacial elastomers. AB - STATEMENT OF PROBLEM: Maxillofacial prosthetic materials should simulate the oral tissues as much as possible and therefore have a similar flexibility and resilience. In light of the oral tissues constantly moving, the dynamic deformation properties of maxillofacial materials would seem the most relevant. PURPOSE: In this study, dynamic mechanical thermal analysis was used to evaluate the deformation properties of five silicone rubber materials used to construct facial prostheses. The technique involves the application of a sinusoidally oscillating stress to a material and analyzes how the material elastically or viscoelastically responds to the stress. The dynamic mechanical thermal analysis can operate at a fixed frequency or range of frequencies over a specific temperature range and also isothermally as a function of time. RESULTS: Cosmesil and A-2186 materials were the most resilient materials and Silbione had the greater energy absorption capacity, which was particularly noticeable at the higher frequencies. Silbione also had a lower shear modulus (G'), which indicated it was more flexible than the other materials. CONCLUSION: The dynamic mechanical thermal analysis proved to be a rapid, reliable, and convenient method for the determination of viscoelastic properties of maxillofacial materials. PMID- 9399195 TI - Clinical effect of full coverage occlusal splint therapy for specific temporomandibular disorder conditions and symptoms. AB - PURPOSE: The purpose of this retrospective study was to evaluate the effect of maxillary full-coverage occlusal splint (stabilization splint) therapy for specific temporomandibular disorders and their symptoms/signs. MATERIAL AND METHODS: This study assessed the outcome of 232 patients who were suffering from chronic pain on movements, joint noise except reciprocal clicking, and difficulty of mouth opening. All were treated with the stabilization splint alone. RESULTS: The total remission rate was 41% and, including those reporting some improvement, the rate was 84%. The presence of displaced disk significantly decreased the success rate. However, the presence or absence of radiographic changes in the temporomandibular joint did not influence the treatment outcome. CONCLUSION: From this study, it is suggested that the stabilization splint therapy may be a useful treatment modality in treatment of temporomandibular disorders, especially for the patients without clinical evidence of displaced disk. PMID- 9399196 TI - Shear bond strength to feldspathic porcelain of two luting cements in combination with three surface treatments. AB - STATEMENT OF PROBLEM: Although selection of clinically reliable bonding system is essential, limited information concerning the combination of durable ceramic bonding and luting agents is available. PURPOSE: This in vitro study was conducted for the purpose of evaluating bond strengths of ceramic bonding materials in conjunction with their initiation and silane-activation modes. MATERIAL AND METHODS: Disk-shaped fired porcelain specimens were air-abraded with alumina, then bonded with six combinations of three silane priming and two luting agents; specimens were also bonded with two luting cements without priming. Shear bond strengths were determined both before and after thermocycling. RESULTS: For the two unprimed control groups, as well as two of the groups bonded with Panavia 21 cement (Clearfil Porcelain Bond and Panavia 21; Panavia Ceramic Primer and Panavia 21) the reduction in bond strengths after thermocycling was remarkable as compared with the corresponding prethermocycling groups (p < 0.05). A dual-cured luting cement (Clapearl DC) used with each of three silane priming materials (Clapearl Bonding Agent, Clearfil Porcelain Bond, and Panavia Ceramic Primer) exhibited consistent shear bond strength greater than 30 MPa after 20,000 thermocycles. CONCLUSION: The above three systems appeared to be useful for the long-term clinical success of feldspathic porcelain restorations. PMID- 9399197 TI - Quantitative study of bacterial colonization of dental casts. AB - STATEMENT OF PROBLEM: Contamination of dental casts can occur if the record bases are improperly disinfected or inadvertently not disinfected during fabrication of a prosthesis. It is essential to develop an effective means of disinfecting dental casts from professional, medical, and legal points of view. PURPOSE: This study determined whether: (1) saliva contamination on the surface of the dental cast contributed to bacterial growth over time and (2) cleaning or disinfecting of dental casts can minimize bacterial growth. MATERIAL AND METHODS: Five dental casts were contaminated with saliva. Each cast was divided into six areas and swabbed at 15, 30, 60, 120, 180, and 240 minutes. Sheep blood agar plates were inoculated and incubated at 37 degrees C for 72 hours. Standardized dental stone cylinders were contaminated with 25 microliters of saliva and treated by rinsing in tap water, scrubbing with soap and tap water, soaking in 2% glutaraldehyde, or as controls with and without saliva contamination (n = 12). The treated dental stone cylinders were placed in individual test tubes containing 2.5 ml of sterile phosphate-buffered solution and a final dilution of 10(-4) was achieved. Sheep blood agar plates were inoculated and incubated at 37 degrees C for 24 hours. RESULTS: Contamination of dental casts did not decrease, even when allowed to sit 4 hours before handling. Results also demonstrated that rinsing saliva-treated stone cylinders for 20 seconds significantly diminished bacterial contamination. Scrubbing with soap and tap water or soaking in 2% glutaraldehyde significantly reduced the bacterial contamination of saliva-treated stone cylinders when compared with rinsing with tap water. CONCLUSION: Bacterial contamination of dental casts can occur and requires an effective method of disinfecting. PMID- 9399198 TI - The effect of environmental pressure changes during diving on the retentive strength of different luting agents for full cast crowns. AB - STATEMENT OF PROBLEM: The effect of pressure cycling on the bond strength of cement luting agents is largely unknown. PURPOSE: This study investigated the effect of pressure cycling on the retention of full cast crowns to extracted teeth. MATERIAL AND METHODS: Sixty extracted single-rooted premolar teeth had full cast crowns cemented, 20 with a zinc phosphate cement, 20 with a glass ionomer, and 20 with a resin cement. After 7 days of storage, each of the teeth in the experimental groups was pressure cycled 15 times from 0 to 3 atmospheres (304 KPa), after which the force required to dislodge the crowns was tested in an Instron testing machine. RESULTS: A significant difference was found (Students t test; p > 0.01) between the force required to remove the crowns in the zinc phosphate control (142.10 +/- 36.42 N) and experimental (15.93 +/- 11.13 N) groups and the glass ionomer cemented control (186.33 +/- 24.33 N) and experimental (91.50 +/- 33.07 N) groups; no difference was found between the resin cemented control (291.15 +/- 78.48 N) and experimental (281.32 +/- 85.43 N) groups. CONCLUSION: This study showed that the retention of full cast crowns to extracted teeth is reduced after pressure cycling if the crowns are cemented with either zinc phosphate cement or glass ionomer cement. Dentists should consider using a resin cement when cementing crowns and fixed partial dentures for patients, such as divers, who are likely to be exposed to pressure cycling. PMID- 9399199 TI - Fabrication of long-span provisional restorations with multiple pontics: a modified method by using a vacuum-formed matrix. PMID- 9399200 TI - A simple antirotational device for implant abutment screws. PMID- 9399201 TI - Customized absorbent wafers for saliva control in implant patients. PMID- 9399202 TI - A simple procedure for boxing elastomeric impressions. PMID- 9399203 TI - Making an impression of a maxillary edentulous patient with gag reflex by pressing caves. PMID- 9399205 TI - Parkinson's disease is not a long-latency illness. PMID- 9399204 TI - Essential tremor: the beginning of a new era. PMID- 9399206 TI - The role of the immune system in neurodegenerative disorders. PMID- 9399207 TI - A gene (ETM) for essential tremor maps to chromosome 2p22-p25. AB - We report the results of linkage analysis in a large American family of Czech descent with dominantly inherited "pure" essential tremor (ET) and genetic anticipation. Genetic loci on chromosome 2p22-p25 establish linkage to this region with a maximum LOD score (Zmax) = 5.92 for the locus, D2S272. Obligate recombinant events place the ETM gene in a 15-cM candidate interval between the genetic loci D2S168 and D2S224. Repeat expansion detection analysis suggests that expanded CAG trinucleotide sequences are associated with ET. These findings will facilitate the search for an ETM gene and may further our understanding of the human motor system. PMID- 9399208 TI - Globus pallidus internus pallidotomy for generalized dystonia. AB - The authors present a young boy with severe generalized dystonia treated with bilateral simultaneous pallidotomy. Microelectrode recordings with the patient under propofol anesthesia showed that the mean discharge rate of globus pallidus internus (GPi) neurons was between 21 and 31 Hz. This contrasts sharply with the mean GPi neuronal firing rates of approximately 80 Hz that are characteristic of Parkinson's disease. The patient had no immediate benefit from surgery, but a progressive improvement in both axial and limb dystonia began within 3 days. The Burke-Fahn-Marsden scores were 75 (maximum possible = 120) at baseline, 52 at 5 days, and 16 at 3 months after surgery. The mechanism of action of pallidotomy for dystonia and the reasons for the delayed and progressive improvement are unknown. Nevertheless, the magnitude of the improvement and the safety of the procedure in this one patient warrant a careful evaluation of pallidotomy for dystonia. PMID- 9399209 TI - Symptoms and duration of the prodromal phase in Parkinson's disease. AB - To investigate the duration of a prodromal phase before the onset of the classic symptoms of idiopathic Parkinson's disease, the authors conducted a retrospective case-control study of 60 patients with Parkinson's disease and 58 age- and sex matched control subjects, covering the decade preceding the onset of classic Parkinson's disease. The symptoms were derived from files of the patients' general practitioners. Compared with control subjects, patients pre-Parkinson's disease had more central nervous system, psychologic, musculoskeletal, and cardiovascular (i.e., autonomic) symptoms. Patients pre-Parkinson's disease also made more visits to general practitioners and medical specialists. The results indicate that the onset of classic parkinsonism is frequently preceded by a prodromal phase lasting from 4-6 years. PMID- 9399210 TI - Postprandial hypotension and parkinsonian state in Parkinson's disease. AB - Abnormal postprandial cardiovascular responses such as postprandial hypotension (PPH) occur in primary autonomic failure and contribute significantly to morbidity. The extent and frequency of PPH and its relationship to the parkinsonian state in idiopathic Parkinson's disease (IPD) is unknown. By studying 20 patients with IPD (without autonomic failure) and 16 age-matched controls after both groups ingested a standard isocaloric balanced liquid meal, we have shown that supine PPH complicates IPD and is related to marked worsening of the parkinsonian state as measured by a cumulative score of tremor, rigidity, bradykinesia, posture, and gait. Furthermore, significant postural hypotension is unmasked that results in postural intolerance due to presyncopal symptoms. Our study indicates that, in patients with IPD, ingestion of a meal may lead to abnormal postprandial cardiovascular responses and aggravation of the parkinsonian stage. The underlying mechanisms are unclear, although vasodilatory gut peptides released in response to food ingestion may be contributory. PMID- 9399211 TI - Differential modification of dopamine transporter and tyrosine hydroxylase mRNAs in midbrain of subjects with Parkinson's, Alzheimer's with parkinsonism, and Alzheimer's disease. AB - The molecular characteristics of midbrain dopamine (DA) neurons have been extensively studied in Parkinson's disease (PD). No such studies of the characteristics of midbrain DA neurons in Alzheimer's disease (AD) or Alzheimer's disease with parkinsonism (AD/Park) have been published. We examined the levels of tyrosine hydroxylase (TH) protein, and the expression of TH and dopamine transporter (DAT) mRNAs, in midbrain neurons of PD, AD, and AD/Park cases. In PD, the loss of TH protein in the ventral tier of the substantia nigra pars compacta (SNpc) of the PD group in accompanied by severe losses in the number of neurons that express TH mRNA and DAT mRNA (74% loss). Remaining neurons show a shift to higher concentrations of TH mRNA but a shift to lower concentrations of DAT mRNA per cell. Hence, there is evidence that compensation in the remaining neurons can elevate concentrations of TH mRNA and lower DAT mRNA. Alternatively, there may be a predilection for a loss of neurons with high levels of DAT mRNA and low TH mRNA levels within the SNpc of PD cases. There was no change in TH protein but an elevation of TH mRNA concentrations per neuron without any change in concentrations of DAT mRNA in the AD group. The AD/Park group did not exhibit changes in the level of TH protein, but showed a small loss (26%) of neurons in the SNpc and a greater loss in other regions of the midbrain (43-53%). Remaining DA neurons showed a marked shift to lower concentrations of DAT mRNA per neuron and a nonsignificant shift in cellular concentration of TH mRNA to higher levels. This is consistent with our previous work showing that with AD/Park there is a significant reduction in the number of DAT sites located on DA terminals in the striatum, but the midbrain neurons have not died. Our results indicate that the differential regulation of mRNAs encoding TH and DAT is similar in the parkinsonian disorders (PD and AD/Park) even though the degree of cell death is very different. This might suggest that compensatory events occur in these DA neurons in AD/Park that are similar to those in PD and that result in differential effects on mRNAs encoding TH and DAT proteins. PMID- 9399212 TI - [123]IBZM binding predicts dopaminergic responsiveness in patients with parkinsonism and previous dopaminomimetic therapy. AB - We investigated the cases of 55 patients with parkinsonism and prior dopaminomimetic therapy in whom the response to this treatment was questionable or reported to be negative. None of these patients had shown motor fluctuations prior to this study. We compared the results of imaging of dopamine-D2 receptors by using [123I]iodobenzamide-single-photon-emission computed tomography (IBZM SPECT) with the improvement in motor signs following a subcutaneous injection of apomorphine and a subsequent increase in oral dopaminomimetic therapy. IBZM-SPECT accurately predicted a positive or negative response to apomorphine in 37 (84%) of 44 patients. The sensitivity/specificity was calculated as 96.3%/ 64.7%. The sensitivity/specificity of IBZM-SPECT for the response to oral treatment with levodopa (L-dopa) was calculated as 100%/75%. After a follow-up period of 2-4 years, 25 patients developed motor fluctuations. All of these patients had normal IBZM binding. Nine developed clinical signs indicating a basal ganglia disorder other than Parkinson's disease. Eight of these nine patients had reduced, and one patient had normal, IBZM binding. We conclude that normal IBZM binding is a useful predictor of a good response to dopaminergic drugs in patients with parkinsonism and a questionable response to previous dopaminomimetic therapy. Reduced IBZM binding seems to exclude a diagnosis of Parkinson's disease, because none of the latter patients clearly benefited from L-dopa and 66.7% developed clinical signs indicating another disorder of the basal ganglia. PMID- 9399213 TI - Proton magnetic resonance spectroscopy in Parkinson's disease and atypical parkinsonian disorders. AB - Proton magnetic resonance spectroscopy (1H-MRS), localized to the lentiform nucleus, was carried out in 12 patients with idiopathic Parkinson's disease (IPD), seven patients with multiple-system atrophy (MSA), seven patients with progressive supranuclear palsy (PSP), and 10 healthy age-matched controls. The study assessed the level of N-acetylaspartate (NAA), creatine-phosphocreatine (Cr), and choline (Cho) in the putamen and globus pallidus of these patients. NAA/Cho and NAA/Cr ratios were significantly reduced in MSA and PSP patients. No significant difference was found between IPD patients and controls. These results suggest an NAA deficit, due to neuronal loss, in the lentiform nucleus of MSA and PSP patients. 1H-MRS is a noninvasive technique that can provide useful information regarding striatal neuronal loss in basal ganglia of patients with atypical parkinsonian disorders and represents a potential tool for diagnosing these disorders. PMID- 9399214 TI - Dependency in Parkinson's disease: a population-based survey in nondemented elderly subjects. AB - Little epidemiological data are available on the dependency status of elderly patients with Parkinson's disease (PD) living in the community. This study assessed the activities of daily living (ADL), the instrumental activities of daily living (IADL), and mobility in a representative sample of elderly nondemented PD subjects (n = 20), compared to a control population (n = 2,697). We found a significantly higher level of dependency in the PD sample based on ADL, IADL, and mobility scales. Half of the PD subjects were dependent for ADL (versus 13.2% for controls), 80% were dependent for IADL (versus 28% for controls), and 20% had their mobility restricted to their home (versus 1.5% for controls). The proportion of PD patients tended to be higher in those more depressed or with more severe motor symptoms. PD patients were not found to be more isolated socially or from family than was the control group and, in any case, dependency seemed not to be associated with isolation. When adjusting for age, sex, depressive symptoms, Mini Mental Status examination score, and dyspnea, PD remained significantly associated with dependency. PD thus constitutes a significant factor of dependency in elderly subjects living at home. Institutionalization occurred over four times more frequently in the PD group than in the general population, but no specific factor of institutionalization was noted. PMID- 9399215 TI - Health-related quality of life in Parkinson's disease: a study of outpatient clinic attenders. AB - OBJECTIVE: To assess the validity and responsiveness of a questionnaire to assess health-related quality of life in Parkinson's disease (PD)--the 39-item Parkinson's Disease Questionnaire (PDQ-39)--and to report problems experienced by patients by means of the questionnaire. METHODS: Patients completed the PDQ-39 and the SF-36 at baseline and 4 months later. At the same assessments, neurologists rated patients with Hoehn and Yahr and Columbia Scales. RESULTS: Evidence for validity of the new questionnaire was observed by agreement of scores with clinical scales at both assessments. Evidence for responsiveness of scales assessing physical function, particularly mobility and activities of daily living, was observed from significant paired t tests for differences between scores at baseline and follow-up, and correlations with patients' retrospective judgments and changes in the SF-36 summary scores. However, there were no significant associations with changes in neurologists' clinical scores. Patients most frequently reported problems of physical function in the PDQ-39. Scores for several dimensions of the PDQ-39 were significantly more favorable than those reported by nonclinic samples of patients with PD. CONCLUSIONS: The PDQ-39 has validity for use among patients attending neurological clinics for treatment of PD. There is also some evidence of responsiveness. The questionnaire identifies problems that are important to patients and that appear to be more commonly experienced by nonclinic attenders. PMID- 9399216 TI - Sexuality in women with Parkinson's disease. AB - There is a renewed interest in sexuality in chronic disease states. Whereas there is some literature on male sexuality in Parkinson's disease (PD), no study has been devoted exclusively to women. We compared 27 women who had PD with community controls matched for age and marital status by using the Brief Index of Sexual Functioning in Women. Approximately 50% of both samples were sexually active. The women with PD were more likely to be dissatisfied with the quality of the sexual experiences. There were significant differences in the two groups with respect to anxiety or inhibition, vaginal tightness, and involuntary urination. Preoccupation with health problems interfering with sex and dissatisfaction with body appearance were also more prevalent in parkinsonian women, but not statistically different from controls. The PD patients were less satisfied with their sexual relationships and with their partners, and were more depressed as a group when compared with controls (Beck Depression Inventory of 11.8 vs 6.3). In both groups, age was associated with significant changes in satisfaction and activity. In summary, qualitative differences exist in the sexual experiences of women with PD compared with controls. PMID- 9399217 TI - Tolcapone added to levodopa in stable parkinsonian patients: a double-blind placebo-controlled study. Tolcapone in Parkinson's Disease Study Group II (TIPS II). AB - The primary objective of this study was to assess the effect of tolcapone on levodopa dosage in parkinsonian patients whose "wearing-off" phenomenon has been controlled with more frequent levodopa dosage. After a 1-week placebo run-in, 97 patients were assigned randomly to receive placebo or tolcapone 200 or 400 mg three times daily (t.i.d.). Levodopa dosage was reduced by -35% on day 1 of study and subsequently retitrated as required. After 6 weeks, the tolcapone groups crossed over to receive the other dose for a further 3 weeks for exploratory purposes. Both tolcapone groups had greater reductions in levodopa dosage than the placebo group at week 6 (not statistically different). The 200-mg t.i.d. group showed greatest improvement in estimated mean scores for all efficacy parameters (p < 0.05 versus placebo for change in Unified Parkinson's Disease Rating Scale Subscale II). Fewer dopaminergic and nondopaminergic adverse events were associated with tolcapone 200 mg t.i.d. than with tolcapone 400 mg t.i.d. The most frequently reported dopaminergic adverse events were nausea, cramps, dyskinesia, and dystonia. The most frequently reported unanticipated adverse event was diarrhea. Tolcapone 200 mg t.i.d. may provide additional benefit to patients with moderately advanced Parkinson's disease with treated "wearing-off" phenomenon. PMID- 9399218 TI - Entacapone enhances levodopa-induced reversal of motor disability in MPTP-treated common marmosets. AB - Oral administration of levodopa (L-dopa) (2.5-25.0 mg/kg) plus carbidopa (12.5 mg/kg p.o.) to MPTP-treated common marmosets produced a dose-related increase in locomotor activity and a corresponding decrease in motor disability. Pretreatment with the peripheral COMT inhibitor entacapone (12.5 mg/kg p.o.) enhanced the intensity and duration of the increase in locomotor activity and the reversal of motor disability produced by a threshold dose of L-dopa (2.5 mg/kg p.o.) plus carbidopa. By contrast, entacapone pretreatment did not potentiate the increased locomotor activity or reversal of motor disability produced by a near-maximal dose of L-dopa (12.5 mg/kg p.o.) plus carbidopa. The effects of entacapone (5.0 25.0 mg/kg p.o.) were dose related, with doses of > 12.5 mg/kg tending to produce less potentiation of L-dopa's effects compared to lower doses. Pretreatment with entacapone (12.5 mg/kg p.o.) without carbidopa caused a short-lasting enhancement of L-dopa's (12.5 mg/kg p.o.) action, whereas pretreatment with carbidopa (12.5 mg/kg p.o.) alone had a more dramatic effect. However, pretreatment with both carbidopa and entacapone produced the greatest overall motor response. In conclusion, entacapone enhances the motor response produced by a low-threshold dose of L-dopa plus carbidopa. However, optimization of both the dose of L-dopa and entacapone appears necessary to obtain the maximal therapeutic response. PMID- 9399219 TI - Constipation in Parkinson's disease: objective assessment and response to psyllium. AB - We evaluated the reliability of patient history and the effect of psyllium on symptoms and colorectal function in 12 patients with Parkinson's disease (PD) and constipation. In all but two, constipation anteceded the development of parkinsonian symptoms. A comparison with prospectively obtained stool diaries confirmed the patients' reported constipation in 7 of the 12 patients. Those patients with confirmed constipation had lower stool weights and reported more straining at stool. Measures of colonic and anorectal function were similar in those who were truly constipated and those who were not. Among those PD subjects with confirmed constipation, psyllium increased stool frequency and weight but did not alter colonic transit or anorectal function. We conclude that prospectively obtained stool diaries should be employed to confirm constipation in PD and that psyllium produces both subjective and objective improvements in constipation related to PD. PMID- 9399220 TI - Effect of chronic oral domperidone therapy on gastrointestinal symptoms and gastric emptying in patients with Parkinson's disease. AB - This study investigated whether domperidone could improve gastrointestinal symptoms in patients with Parkinson's disease who were receiving levodopa therapy. A total of 11 patients were studied. Following a baseline gastric emptying test, patients were treated with a starting dose of domperidone 20 mg p.o. q.i.d. A follow-up gastric emptying test was repeated at least 4 months after starting domperidone therapy. At the beginning and at each 3-month follow up visit, symptoms of nausea, vomiting, anorexia, abdominal bloating, heartburn, regurgitation, dysphagia, and constipation were evaluated and scored on a scale of 0-3. The overall mean follow-up period was 3 years. Compared with their baseline evaluation, patients experienced a significant improvement in all symptoms (p < 0.05) except dysphagia and constipation. Gastric emptying of an isotope-labeled solid meal was significantly faster, with a baseline result of 60.2 +/- 6.4% retention of isotope 2 h after the meal compared with 37.0 +/- 2.2% retention during domperidone therapy (p < 0.05). Patients' global assessment of Parkinson's disease remained stable or improved. Serum prolactin was elevated in all patients after domperidone therapy (p < 0.05). Domperidone therapy significantly reduces upper gastrointestinal symptoms and accelerates gastric emptying of a solid meal, but does not interfere with response to antiparkinsonism treatment. PMID- 9399221 TI - Interlimb coordination deficits in patients with Parkinson's disease during the production of two-joint oscillations in the sagittal plane. AB - Two-limb coordination patterns involving cyclical flexion-extension movements, performed in the same or in different directions, were studied in patients with Parkinson's disease and a group of elderly subjects. The three patterns referred to the homologous (both arms or legs), homolateral (right or left arm and leg), and heterolateral (right arm and left leg or vice versa) limb segment combinations that were performed in the sagittal plane from a seated position. Findings revealed that interlimb coordination deficits were evident in patients with Parkinson's disease. Moreover, mean cycle duration and its variability were increased, particularly during the production of nonhomologous limb movements in different directions. These temporal findings suggest that movement slowness was not a primary consequence of an intrinsic inability to move the limb segments at the required speed but rather reflected an intentional strategy to cope with the complexity of the coordination pattern. Finally, movement amplitude was substantially smaller and more variable in patients with Parkinson's disease, suggestive of hypometria during the production of these cyclical tasks. PMID- 9399222 TI - Clinical expression of essential tremor: effects of gender and age. AB - Essential tremor (ET) is considered to be a monosymptomatic disorder consisting primarily of postural hand tremor. Nevertheless, clinical expression can vary based on the body region affected by tremor and the coexistence of other neurologic signs, such as tandem gait disturbance. We conducted a two-part study to test the hypothesis that variability in ET clinical expression is influenced by gender and age. In part 1, we examined a large ET clinical database (n = 450), comparing ratings of postural hand and head/voice tremor based on gender. Head/voice tremor was significantly more frequent and more severe among female ET patients; men had more severe postural hand tremor. In part 2, ET patients (n = 40) had significantly more missteps when tandem walking in comparison to age matched controls. Poor tandem walk in ET cases was associated with more advanced age, but not gender, disease duration, or ratings of postural hand or head/voice tremor. We conclude that gender influences the body region most affected by ET possibly through the effects of the sex chromosomes or hormones. Ataxia (tandem gait difficulty) is common in ET and may be an accentuation of cerebellar dysfunction due to aging. PMID- 9399223 TI - Agreement among movement disorder specialists on the clinical diagnosis of essential tremor. AB - Even though essential tremor (ET) is the most prevalent movement disorder, there has been little agreement in the neurologic literature regarding diagnostic criteria for ET. The authors attempted to determine the extent and source of agreement and disagreement among neurologists regarding diagnostic criteria for clinically definite ET. The authors designed and mailed a semistructured questionnaire to 160 neurologists who specialize in movement disorders in 24 countries. The questionnaire included three sections: a list of inclusion criteria, a list of exclusion criteria, and a list of potential clinical scenarios (for example, isolated site-specific tremor and primary orthostatic tremor). The questionnaire was completed by 98 (61%) of 160 targeted neurologists. There was greater consensus regarding features considered unnecessary inclusion criteria for clinically definite ET (extent of disability, disease duration, and positive family history) than for those considered necessary inclusion criteria (postural versus action tremor). With regard to exclusion criteria, there was some consensus in terms of the presence of Parkinson's disease, dystonia, history of hyperthyroidism or concurrent use of tremor-inducing medications, and cerebellar signs. The majority of neurologists would diagnose ET in the setting of isolated head or voice tremor. There are areas of both consensus and divergence among neurologists with regard to diagnostic criteria for ET. The choice of diagnostic criteria may vary depending on the intended use of the criteria (that is, clinical versus genetic studies). Hopefully, this study will foster further discussion to achieve a more general consensus. PMID- 9399224 TI - Cerebellar axial postural tremor. AB - Three cases are presented with a predominantly axial postural tremor, without visible palatal tremor. Tremor varied in frequency between 3 and 10 Hz, often jumping from one frequency to another in this band. All three patients had evidence of cerebellar pathology. Cases 1 and 2 developed tremor in the setting of a late-onset cerebellar degeneration and after excision of a right cerebellar haemangioblastoma, respectively. Etiology was unclear in Case 3. Nevertheless, this patient had a cerebellar dysarthria. The tremor was similar to that sometimes seen in conjunction with palatal tremor, and EMG studies in Case 3 demonstrated a subclinical modulation of palatal muscle activity simultaneous with the truncal tremor. It is suggested that an axial postural tremor may be due to pathology of the cerebellum and its outflow pathways, despite the absence of clinically apparent palatal tremor. PMID- 9399225 TI - Validity of long-term electromyography in the quantification of tremor. AB - We previously developed a method of tremor quantification using long-term electromyography registration of antagonistic forearm muscles that is reliable, sensitive, and specific for pathologic tremors. The present study demonstrates that tremor occurrence as measured by this method correlates well with clinical parameters of tremor in idiopathic Parkinson's disease (PD) and essential tremor (ET) (subscores of the Unified Parkinson's Disease Rating Scale for PD, tremor rating according to Bain et al. for ET). We conclude that the method is a valid and objective means of tremor quantification in PD and ET. PMID- 9399227 TI - Frequency of familial inheritance among 488 index patients with idiopathic focal dystonia and clinical variability in a large family. AB - Idiopathic torsion dystonia is characterized by involuntary twisting movements and postures. One molecularly defined form with generalized dystonia has been shown to be autosomal dominantly inherited with reduced penetrance in chromosome 9q34.1, especially in Ashkenazi Jewish families, while other generalized families from Europe and families with other subtypes of dystonia have been excluded from linkage to this locus. Genealogical studies suggest that the much more frequent focal dystonia follows an autosomal dominant inheritance with reduced penetrance as well. For our study, 488 patients with focal dystonia, without a tendency for generalization, were interviewed for their family history. Evidence for hereditary disposition was found in 88 individuals. In a second step, all available family members of 17 of the 488 index patients (chosen for cooperation) were clinically examined. Objective diagnosis of affected relative was established in 13 families, whereas only 4 of the 17 index patients had previously admitted a positive family history. Furthermore, a large three generation family with focal dystonia linked to chromosome 18p (linkage data described elsewhere) was identified. The familial pattern of all reported families is compatible with autosomal dominant inheritance with reduced penetrance. Assessment only on patients' report leads to underestimation of the frequency of familial idiopathic focal dystonia. PMID- 9399226 TI - Analysis of action tremor and impaired control of movement velocity in multiple sclerosis during visually guided wrist-tracking tasks. AB - We investigated the relationship between action tremor (AT) and impaired control of movement velocity (MV) in visually guided tracking tasks, in normal subjects and in patients with multiple sclerosis (MS) with or without motor deficits. The effects of withdrawing visual feedback of either the target or the cursor were then investigated. Visually cued simple reaction times (SRTs) were also measured. The effects of thalamotomy on motor performance in these tasks were evaluated in seven patients. In the MS patients with tremor, there was no correlation between AT and impairment in control of MV, but the latter was highly correlated with an increased delay in SRT. Withdrawal of visually guiding cues increased the error significantly in MV, but reduced AT by approximately 30% in magnitude. Frequency analysis indicated that the AT had two components: (a) non-visual-dependent, oscillatory movements, mainly at 4 Hz; and (2) visual-dependent, repetitive movements, with significant power at 1-2 Hz. Thalamotomy significantly reduced AT but hardly improved accuracy in MV. These results suggest that visual feedback of a spatial mismatch signal may provoke a visually dependent repetitive movement contributing to AT. Conduction delays along either the cortico-cerebello-cortical or the proprioceptive pathways and impaired working memory caused by MS may be responsible for the movement disorders in these patients. PMID- 9399228 TI - Paroxysmal exercise-induced dystonia: eight new sporadic cases and a review of the literature. AB - We report eight new sporadic cases of paroxysmal dystonia induced by prolonged or sustained exercise and review an additional seven previously reported cases. The attacks in our patients lasted from a few minutes to up to 2 h, and patient age at onset ranged from 2 to 30 years. Four of the eight patients had hemidystonic attacks, both legs were involved in two other cases, and the remaining two patients had involvement of the right foot only. We propose that such cases should be classified as paroxysmal exercise-induced dystonia. PMID- 9399229 TI - DYSBOT: a single-blind, randomized parallel study to determine whether any differences can be detected in the efficacy and tolerability of two formulations of botulinum toxin type A--Dysport and Botox--assuming a ratio of 4:1. AB - BACKGROUND: Elston and Russell discovered a difference in the biological potency of the English formulation of botulinum toxin type A or BTX-A (Dysport) and the American formulation (Botox). Potency of both is expressed in LD50 mouse units, but because of assay differences, these units are not equivalent. Since the first warning by Quinn and Hallet on the clinical importance of this issue, it has been impossible to reach a consensus on the conversion factor for the potency of these formulations. OBJECTIVE: To test the hypothesis that the conversion factor for the clinical potency of Dysport to Botox is approximately 4:1. DYSBOT is an acronym that results from adding "DYS" from Dysport with "BOT" from Botox. PATIENTS AND METHODS: DESIGN: A single-blind, randomized, parallel comparison. A total of 91 patients with blepharospasm or hemifacial spasm were randomized to treatment with Dysport or Botox using a fixed potency ratio of 4:1. Clinical evaluations: The patients were evaluated at baseline (day of the treatment). 1 month after treatment, and whenever the effect was judged to be fading. Objective and functional rating scales were used as quantitative measures of the change in clinical status. Adverse reactions were collected using a systematic questionnaire. RESULTS: Using this ratio between products, both Dysport and Botox groups produced similar clinical efficacy and tolerability. For patients showing a positive response without the need of a booster, the duration of effect was 13.3 +/- 5.9 weeks for the Dysport group and 11.2 +/- 5.8 weeks for the Botox group. Of 48 patients, 11 (23%) needed booster treatment in the Dysport group compared with five (12%) of 43 in Botox group. Adverse events were noted in 24 (50%) of 48 patients in the Dysport group and 20 (47%) of 43 of the Botox-treated group. CONCLUSIONS: Using a 4:1 conversion ratio for Dysport and Botox, similar results were obtained for the two treatments in an appropriately powered study, suggesting that this conversion factor is a good estimate of their comparative clinical potencies. PMID- 9399230 TI - Kinematic analysis of articulatory movements in central motor disorders. AB - The various components of the central motor system are expected to play a similar role in speech production and in upper limb control. Slowed articulatory performance, therefore, must be expected in disorders of the corticobulbar tracts, cerebellum, and basal ganglia. Using an optoelectronic device, the present study recorded lower lip trajectories during production of sentence utterances in patients with Parkinson's disease (PD), Huntington's disease (HD), cerebellar atrophy (CA), and pseudobulbar palsy (PB). The various subject groups showed a similar range of overall motor disability. Patients with CA and PB exhibited slowed movement execution in terms of a reduced ratio of peak velocity to maximum amplitude ("stiffness"). In contrast to upper limb motor control, the lip excursions showed an uncompromised shape of velocity profiles. Two different patterns emerged in HD. A single patient suffering from the akinetic-rigid Westphal variant of this disease had articulatory hypometria, whereas the remaining subjects showed significant bradykinesia under increased temporal demands, concomitant with normal movement amplitudes. The PD patients had unimpaired velocity-displacement relationships. Presumably, biomechanical constraints such as the rather small excursions of articulatory lower lip gestures or the scarce spindle supply of facial muscles account for the observed discrepancies between upper limb and speech motor control in PD. PMID- 9399231 TI - Analysis of blink rate patterns in normal subjects. AB - The present study measured the normal blink rate (BR) variations in relation to behavioral tasks in 150 healthy volunteers (70 males and 80 females; aged 35.9 +/ 17.9 years, range 5-87 years). The subjects were videotaped in a standard setting while performing three different tasks: resting quietly, reading a short passage, talking freely. The mean BR was computed during each task; the data were compared by means of analysis of variance and Student's t tests. Mean BR at rest was 17 blinks/min, during conversation it increased to 26, and it was as low as 4.5 while reading. As compared with rest, BR decreased by -55.08% while reading (p < 1 x 10(-15)) and increased by 99.70% during conversation (p < 1 x 10(-9)). As compared with reading, BR increased during conversation by 577.8% (p < 1 x 10( 17). The distribution curves were highly reproducible in each task. The best curve fit was represented by a log-normal distribution, with the upper tail of each curve having a normal distribution. Eye color and eyeglass wearing did not influence BR. Women had higher BR than men just while reading. No age-related differences were found. The most common BR pattern was conversation > rest > reading, which occurred in 101 subjects (67.3%); 34 subjects (22.7%) had the pattern rest > conversation > reading; 12 (8.0%) had the pattern conversation > reading > rest. This study identified three normal behavioral BR patterns and showed that BR is more influenced by cognitive processes than by age, eye color, or local factors. The present findings provide a normal reference for the analysis of BR in movement disorders such as dystonia or tics. PMID- 9399232 TI - Parkinsonism associated with acute intracranial hematomas: an [18F]dopa positron emission tomography study. AB - We present the case of a 36-year-old woman with a right temporal hematoma and an overlying subdural hematoma following rupture of a right middle cerebral artery aneurysm. Three weeks after recovering consciousness, she developed a levodopa responsive parkinsonian syndrome involving the right limbs. A year after the vascular event, she reported subjective improvement in her parkinsonism, which has remained stable since. 18F-dopa positron-emission tomography showed a marked reduction of uptake in the left putamen, raising the possibility that the intracranial hemorrhage unmasked latent Parkinson's disease. To the best of our knowledge, this is the first case of parkinsonism associated with spontaneous acute intracerebral and subdural hematomas. PMID- 9399233 TI - Efficacy of a patient-training videotape on motor fluctuations for on-off diaries in Parkinson's disease. AB - Patient on-off diaries are used in clinical trials, but a method to assure agreement between patient and examiner has never been developed. We tested whether a patient-teaching tape increased the rate of agreement between patient diary ratings and simultaneous neurologic assessment by a trained professional. A total of 32 consecutive patients who had Parkinson's disease with motor fluctuations independently completed a 4-h on-off diary (nine ratings) at the same time as an examiner. Those with < 80% agreement with the examiner (n = 20) were randomized to view either a training tape that showed motor fluctuations (experimental group) or-another videotape of general patient educational material (control group). All patients then underwent the same 4-h assessment of motor fluctuations. To test for long-term retention, they returned 1 month later and, without reviewing the videotape, underwent a final 4-h correlation assessment. After the training tape, the experimental group showed significant improvement, whereas the control group showed no improvement. Furthermore, another month later, the improvement in the experimental group was retained. Based on these findings, we suggest that future clinical trials assessing motor fluctuations incorporate this tape into their basic methodology. PMID- 9399234 TI - Spinal myoclonus induced by an intrathecal catheter. AB - We report a case of spinal myoclonus induced by the tip of an intrathecal catheter in a 35-year-old patient with severe, adult-onset, generalized dystonia of unknown cause, treated for 2 years using intrathecal baclofen. One month after a falling episode, the patient developed focal myoclonus of the right proximal leg whenever she stood up from a seated position. The electrophysiologic recordings were compatible with spinal segmental myoclonus, originating at a focus corresponding to the L2-S2 segments. At this site, the tip of the intrathecal catheter was demonstrated by myelography to be in close proximity to the nerve roots and conus medullaris. The myoclonus resolved promptly once the catheter tip was withdrawn. We review the literature on spinal myoclonus and discuss the possible mechanisms of spinal myoclonus pertaining to the present case. This report represents an unusual complication of intrathecal catheter systems that, if recognized, can lead to prompt therapeutic intervention. PMID- 9399236 TI - Adult-onset tics associated with peripheral injury. AB - We report the cases of two patients with adult-onset, simple, nonvarying tic disorder that commenced after a peripheral (non-CNS) injury. The first patient is a 38-year-old man who suffered a right facial injury when his car fell off its jack while he was working underneath. Bilateral facial twitching began hours after the trauma and was characterized as a sniffinglike gesture. The movements waxed and waned, were suppressible, and were associated with a premonitory sensation. The tics remitted after 9 months but still recur occasionally under stressful situations. The second patient is a 34-year-old man with a 3-year history of abrupt, rapid head-turning movements that began 12 months after a motor vehicle accident in which he injured his neck. The tics continue to wax and wane, are suppressible, and are associated with an urge. Neither patient suffered a head injury or had a family history of Tourette syndrome. Based on the clinical and historical features of these patients, the temporal relationship between the trauma and onset of tics, and the occurrence of tics only in the traumatized region, a causal relationship is possible. These may represent the first reported cases of tic disorder in association with peripheral injury. The mechanism by which the tic disorder resulted from the peripheral injury is unclear, but these patients might have been susceptible individuals and the trauma acted as a trigger. PMID- 9399237 TI - Spinal rigidity following acute myelitis. AB - A patient with a 2(1/2)-year history of painful spasms and rigidity of both lower limbs is described. Symptoms began after an episode of acute myelitis. The spasms -which were spontaneous and stimulus sensitive and occurred on voluntary action- involved the repetitive grouped discharge of motor units. Continuous motor unit activity was present at rest in the muscles of both legs, and cutaneomuscular reflexes were abnormal. This patient is similar to those recently reported as having stiffness and spasms of the legs due to a possible chronic spinal interneuronitis and provides further evidence that this kind of movement disorder may be caused by spinal cord pathology. PMID- 9399235 TI - Cortical Myoclonus in Huntington's disease associated with an enlarged somatosensory evoked potential. AB - We report the electrophysiologic findings of myoclonus in a patient with Huntington's disease (HD). This patient was studied postoperatively after a bilateral fetal cell transplant in his striatum. Incomplete transient improvement was seen in the myoclonus, followed by gradual deterioration. The myoclonus itself had a cortical correlate and was associated with an enlarged somatosensory evoked potential (SEP), consistent with the presence of cortical reflex myoclonus. An enlarged SEP has not been previously reported in myoclonus associated with HD. The postulated mechanisms for myoclonus, when it occurs in HD, have differed in the literature. The reason for the transient improvement of the myoclonus following transplantation is unclear, but this case raises the possibility that basal ganglia circuits may modulate cortical myoclonic activity. PMID- 9399238 TI - Botulinum toxin A improves muscle spasms and rigidity in stiff-person syndrome. AB - We studied the effect of botulinum toxin A (BTA) on painful muscular spasms and rigidity in two bedridden patients with clinical, electrophysiologic, and immunologic evidence of stiff-person syndrome. We injected BTA or saline solution into several limb muscles with both the rater and patient blinded to the order of the injections. A physician, unaware of the treatment order, used an objective rating scale for rigidity and spasm frequency scale and independently assessed the treatment results. BTA administration significantly reduced rigidity and stopped the spasms in all limbs. Following BTA injection on one side, the spasm frequency decreased bilaterally possibly because of the spread of hematogenous toxin. PMID- 9399240 TI - The shaking palsy, the first forty-five years: a journey through the British literature. AB - The authors examined the British medical literature published in the 45-year period following Parkinson's treatise on the shaking palsy to determine the number and type of references to the shaking palsy or paralysis agitans during this particular period. Several sources suggest that Parkinson's 1817 treatise on the shaking palsy received little immediate attention in his native country, England, and that not until 1861, in France, did Charcot began to elucidate the clinical features of this entity, separating it from other neurologic disorders (for example, multiple sclerosis). A review of the British medical literature from the 45-year-period 1817-1861 revealed a number of references to paralysis agitans, including those by Cooke (1820), Good (1824 and 1829), Elliotson (1827, 1829, 1830, 1831, and 1833), Gowry (1831), anonymous (1832), Todd (1833), Watson (1836), Gibson (1839), Hall (1838 and 1841), Thompson (1842), Graves (1843), Birkett (1853), Paget (1855), and Reynolds (1855). Many of these did not report new or personally observed cases, did not separate Parkinson's disease from other disease entities characterized by both "shaking" and "palsy" (for example, tonic clonic seizures), or misattributed motor signs to dysfunction of the pyramidal system rather than an extrapyramidal system (that is, attributing bradykinesia or rigidity to weakness). Although there were several references to "shaking palsy" in the early- to mid-19th-century British medical literature, there were few original case reports of Parkinson's disease. This may have contributed to the fact that during this period little was added to the original observations made by Parkinson in 1817. In particular, the separation of bradykinesia and weakness did not become apparent until later work by the French. PMID- 9399239 TI - Idiosyncratic adverse reactions to intramuscular botulinum toxin type A injection. AB - Three cases of adverse reactions to repeated intramuscular botulinum toxin A (BTA) injections are described: a persistent rash on the face at the site of injection, a localized anaphylactic reaction following BTA injection into one leg, and bilateral ptosis repeatedly following BTA injection into neck muscles. The mechanisms for these idiosyncratic adverse responses are not known. PMID- 9399241 TI - A historical case of probable corticobasal degeneration? AB - In 1925, Jean Lhermitte and colleagues published a work on spatial representation in apraxic patients, based on two illustrative cases. One of these displayed a clinical picture that would nowadays be clinically classified as probable corticobasal degeneration. PMID- 9399242 TI - Progressive cognitive decline with truncal/limb ataxia and ballistic movements. PMID- 9399243 TI - Botulinum toxin injections to one leg alleviate freezing of gait in a patient with Parkinson's disease. PMID- 9399244 TI - Unilateral parkinsonism following a large infarct in the territory of the lenticulostriate arteries. PMID- 9399245 TI - Remitting parkinsonism as a symptom of multiple sclerosis and the associated magnetic resonance imaging findings. PMID- 9399246 TI - Left arm monoballism as a relapse in multiple sclerosis. PMID- 9399247 TI - Spasmodic torticollis associated with multiple sclerosis: report of two cases. PMID- 9399248 TI - Painful tonic spasms and pure motor hemiparesis due to lacunar pontine infarct. PMID- 9399249 TI - Unilateral postural and action tremor resulting from thalamic toxoplasmosis in a patient with acquired immunodeficiency syndrome. PMID- 9399250 TI - Delayed onset postural tremor caused by parietal lesion. PMID- 9399251 TI - Sustained effect of high-dose intrathecal baclofen in primary generalized dystonia: a 2-year follow-up study. PMID- 9399252 TI - Successful treatment of fluoxetine-induced dystonia with low-dose mianserin. PMID- 9399253 TI - Apraxia of eyelid closure accompanied by denial of eye opening. PMID- 9399254 TI - Tardive dyskinesia in a neuroleptic-naive patient with bipolar-I disorder: persistent exacerbation after lithium intoxication. PMID- 9399255 TI - Italian buckwheat (Fagopyrum esculentum) starch: physico-chemical and functional characterization and in vitro digestibility. AB - A study on the physico-chemical properties and structure together with the evaluation of starch digestibility was carried out on starch isolated from buckwheat (Fagopyrum esculentum) cultivated in different Italian areas. Results showed that buckwheat samples analysed were different among them and from wheat starch used as reference. Buckwheat granules were polygonal in shape and had a smaller diameter than the wheat starch granule. The starch obtained from buckwheat had a higher swelling power than the wheat one, probably as a consequence of the wheaker but more extensive bonding forces in the granule. During cooling, buckwheat samples showed a good paste stability. PMID- 9399256 TI - Available iron and zinc in major lean meat cuts and their contribution to the recommended trace element supply in Switzerland. AB - The objective of the present study was to analyze iron and zinc in major lean meat cuts in order to estimate the contribution of the average lean meat consumption in Switzerland (1995) for these trace elements. Iron, heme iron and zinc contents were analyzed in following muscles: pork (longissimus dorsi muscle and shoulder), beef (longissimus dorsi muscle and shoulder), veal (longissimus dorsi muscle) and chicken (breast and thigh). Beef and pork shoulder were the best sources of iron, heme iron and zinc. Pork and veal longissimus dorsi muscle and chicken were relatively poor in these trace elements. With an average daily lean meat consumption of 105 g, iron and zinc intake were about 1.1 mg/d and 3.8 mg/d, respectively. Recommendations for daily iron intake were met to 11% (men) and 7% (women) and for zinc to 25% (men) and 32% (women). Applying a modified Monsen model, the requirements for absorbed iron were met in the range of 10-30% and 7-20% for men and women, respectively. Taking into account a zinc absorption rate from meat of about 20-36%, the daily requirements for absorbed zinc were covered to 32-56%. In conclusion, the average amount of lean meat as consumed in Switzerland was high enough to be an important source of available iron and zinc, particularly for people with low iron and zinc status. PMID- 9399257 TI - Production of iron-fortified bread employing some selected natural iron sources. AB - Iron fortification of wheat breads is the optimal approach for reducing the high prevalence of iron deficiency in wheat-eating developing countries as in Egypt. The effectiveness of some natural iron-fortificants for potential use in Egyptian bread was tested. Defatted soybean flour, soybean flour, soybean hull flour, molasses and fenugreek flour at different levels besides FeSO4.7H2O as standard were separately incorporated in the wheat flour dough. Dough characteristics were studied using a Brabender Farinograph, where addition of such fortificants improves significantly (p < 0.05) the water absorption, development time, dough stability and dough weakening. Good breads with high nutritive value, excellent crust color, crumb grain and high overall acceptability were produced by adding either 10% defatted soybean flour, 5% soybean hull flour, 2% molasses or 4% fenugreek flour. Rat feeding trials have shown a good haematological response, i.e. higher haemoglobin (Hb) and haematocrit (Hct) values gained. Additional work is needed to identify other fortification options and to develop targeted fortification programes that will supply iron to all segments of a population in greatest need. PMID- 9399258 TI - Influence of vegetarian and mixed nutrition on selected haematological and biochemical parameters in children. AB - To evaluate the health and nutritional status of children with two different nutritional habits, the authors examined 26 vegetarians (lacto- and lacto-ovo; an average period of vegetarianism 2.8 years) and 32 individuals on mixed diet (omnivores) in the age range 11-14 years. Vegetarian children had significantly lower erythrocyte number as well as reduced levels of haemoglobin and iron compared to omnivores. The average level of iron did not reach the lower limit of the physiological range and hyposiderinemia was found in 58% of vegetarians vs 9% of omnivores. Reduced iron levels were observed in spite of increased intake of vegetable iron sources and vitamin C (which facilitates the conversion to ferro form). This reduction can be attributed to the absence of animal iron sources with high utilizability and to lower iron utilization in the presence of phytic acid (higher intake of grains compared to omnivores). The incidence of hypoalbuminemia and hypoproteinemia in vegetarian children was 38 and 12%, respectively, compared to 0% in omnivores. The protein mixture from milk, eggs and vegetable sources is complete, but vegetarian children had significantly reduced intake of milk and dairy products. Favourable lipid and antioxidant parameters in vegetarian children reflect the optimal nutrition composition with respect to the prevention of free radical diseases. Such a nutrition results in significantly lower levels of cholesterol and LDL-cholesterol compared to omnivores and significantly higher and over threshold values of essential antioxidants--vitamin C, vitamin E/cholesterol (more effective protection against LDL oxidation), beta-carotene, vitamin A. PMID- 9399259 TI - More questions than answers. PMID- 9399260 TI - Article on breast cancer risk and induced abortion concerns readers. PMID- 9399261 TI - Multispecialty clinics enhance consultation and care. PMID- 9399262 TI - Critical pathway improves outcomes for patients with sickle-cell disease. PMID- 9399263 TI - Laying the foundation: leadership council operationalizes values and goals. PMID- 9399264 TI - Ambulatory nurses establish group practices to improve efficiency and satisfaction. PMID- 9399265 TI - Primary nursing improves access to and quality of care in screening clinics. PMID- 9399266 TI - Linkage key to rural outreach program success. PMID- 9399267 TI - Go light your world. PMID- 9399268 TI - The nurse as coach: a conceptual framework for clinical practice. AB - PURPOSE/OBJECTIVES: To operationalize a professional educational counseling model for nurses that derives from the client's frame of reference and adds to the client's behavioral management of the impact of cancer, including self-care skills and cognitive control. DATA SOURCES: Published literature and four years of clinical experience with 84 couples in which coaching behavior was applied in home-based intervention sessions. DATA SYNTHESIS: Nurse coaching behavior includes six dimensions. Attending to the Story, Encircling the Experience, inviting the Work, Exploring Solutions, Anchoring the Skill, and Setting Up Success. CONCLUSIONS: Nurse coaching behavior is designed to facilitate the cognitive emotional processing of the cancer experience and to add to the patient and family member's repertoire of behavioral self-care and self-management skills. Future research is needed to evaluate the processes and outcomes of nurse coaching behavior when working with patients and family members experiencing cancer. IMPLICATIONS FOR NURSING PRACTICE: Nurse coaching provides a practice framework that complements patient teaching and supportive therapy as a method for enhancing self-care and self-management behavior for people with cancer and their family members. PMID- 9399269 TI - Addressing issues for early detection and screening in ethnic populations. AB - PURPOSE/OBJECTIVES: To describe how multicultural knowledge and skills are applied in culturally competent practice to develop and deliver available, accessible, acceptable, and appropriate programs in early cancer detection and screening programs in ethnic communities. DATA SOURCES: Literature and clinical community practice reports. DATA SYNTHESIS: Successful community screening programs can be conducted within ethnic minority populations. Practitioners must tailor care based on salient cultural differences of the population of focus. Culturally based practice results in the (a) increased ability to think critically in community and individual assessments, (b) development of more accurate program plans and designs, and (c) increased likelihood that appropriate outcomes will be used in the evaluation of care. CONCLUSIONS: Greater cultural competence increases the accuracy of care and thereby its effectiveness, efficiency, and success in providing acceptable and optimal programs. IMPLICATIONS FOR NURSING PRACTICE: The first step in a cultural assessment is to know one's own beliefs and attitudes. The second step, conducted in parallel, requires knowledge about the groups that make up the patient and staff populations within the practice setting and then integrating those perspectives into practice. Practitioners then can begin the third step of negotiation of all stages of the project with members of the various communities on an equal basis that recognizes the expertise and integrity of all parties. PMID- 9399270 TI - Determinants of exercise during colorectal cancer treatment: an application of the theory of planned behavior. AB - PURPOSE/OBJECTIVES: To examine determinants of exercise during colorectal cancer treatment using the theory of planned behavior. DESIGN: A retrospective survey. SETTING: Cancer Registry of Alberta, Canada. SAMPLE: Randomly selected survivors of colorectal cancer (N = 110) diagnosed between 1992 and 1995 who had undergone adjuvant therapy. Participants' ages ranged from 26 to 77 years (mean = 61 years; SD = 11), 63% were male, and 85% were disease stage II or III. METHODS: Initial open-ended elicitation questionnaire to determine salient beliefs, a mailed main questionnaire, a postcard reminder one week later, and a second questionnaire three weeks later. Exercise was assessed by the Godin Leisure Time Exercise Questionnaire. MAIN RESEARCH VARIABLES: Exercise during cancer treatment, intention, perceived behavioral control, attitude, subjective norm, and salient beliefs. FINDINGS: Exercise during cancer treatment was determined by intention and perceived behavioral control. Intention was determined solely by attitude. Salient beliefs about exercise were different for patients with cancer as compared to a healthy population. CONCLUSIONS: The theory of planned behavior may be a viable framework on which to base interventions to promote exercise in patients with colorectal cancer. IMPLICATIONS FOR NURSING PRACTICE: Oncology nurses need to have an understanding of motivational factors related to exercise during cancer treatment to be able to assist patients with cancer to initiate and maintain exercise. PMID- 9399271 TI - Oncology nurses' practices of assisted suicide and patient-requested euthanasia. AB - PURPOSE/OBJECTIVES: To provide reliable and valid empirical data related to New England Oncology Nursing Society (ONS) members' self-reported practices of assisted suicide and patient-requested euthanasia. Analysis focused on the nurses' practices, a comparison of their practices to a similar sample of oncology physicians, and their use of the healthcare team. DESIGN: Quantitative survey. SETTING: New England region of the United States. SAMPLE: 600 ONS members surveyed by mail, 441 of whom responded (74% return rate). Only nurses who worked at least 20 hours per week, were ONS members for at least one year, and worked with adult patients with cancer were included. METHODS: Replication and extension of a survey of oncology physicians. MAIN RESEARCH VARIABLES: Frequency of requests for and responses to patient requests for assisted suicide and euthanasia and the use of the healthcare team in response to these requests. FINDINGS: More physicians than nurses assisted their patients' suicides (11% versus 1%). However, nurses were more likely than physicians to have performed patient-requested euthanasia (4% versus 1%). Nurses frequently consulted with others--particularly physicians--about patient requests for assistance with death but rarely with one another including nursing supervisors. CONCLUSIONS: The relative number of healthcare professionals (physicians or nurses) who admit to hastening a patient's death is small. Nurses in this study received fewer requests to perform euthanasia than physicians, but they performed patient requested euthanasia four times more frequently than physicians. Professional affiliation appears to be one factor in determining whether or not a patient's request for assistance with death will be granted. IMPLICATIONS FOR NURSING PRACTICES: The policy debate about professional roles in actions that end the lives of patients must be extended beyond physicians to include nurses. Nurses must take an active role in the discussion and definition of acceptable practice at the end of life. PMID- 9399273 TI - Primary caregiver perceptions of intake cessation in patients who are terminally Ill. AB - PURPOSE/OBJECTIVES: To explore the meaning of nutrition cessation in adult in home hospice patients with cancer as described by women primary caregivers during the first year of bereavement. SETTING: In-home hospice in the northeastern region of the United States. SAMPLE: Twelve English-speaking adult women who had cared for patients who were terminally III with cancer who ceased oral intake. DESIGN: Qualitative phenomenologic inquiry. METHOD: Verbatim written transcripts of semistructured interviews were studied line by line to identify themes. Shared themes emerged through ongoing comparison across cases. FINDINGS: Within the framework of transition, seven essential themes emerged: The Meaning of Food, Caregiver as Sustainer, Concurrent Losses, Personal Responses, Ceasing to Be- "Starved to Death," Being Bereaved--The Meaning Now, and Paradox. Patient changes in intake patterns and caregiver actions to encourage intake were described. Decreasing intake led to ongoing and spiraling losses. Caregiver intake patterns also changed. CONCLUSIONS: Caregivers believed that patient-nutrition cessation was naturally occurring and not physically painful. IMPLICATIONS FOR NURSING PRACTICE: Sensitized nurses can look for the presence of the phenomenon in cancer caregiving families and open dialogue. Anticipatory guidance can serve to normalize the situation and ease the transition. Future research should focus on what nurses know and what they share with families regarding intake cessation. Research with caregivers could address values clarification, decision making, knowledge needed for caregiving, the perceptions of caregiving men, and caregivers of diverse cultures. More research conducted with patients ceasing intake is needed to determine whether and to what degree patients suffer. PMID- 9399272 TI - Health promotion and early detection of cancer in older adults: assessing knowledge about cancer. AB - PURPOSE/OBJECTIVES: To identify the knowledge level concerning cancer in older Canadian adults. DESIGN: Descriptive. SETTING: Urban community in Canada. SAMPLE: Convenience sample of 513 adults over the age of 55 (72% female; 68% born in countries other than Canada). METHODS: A self-report questionnaire, the Cancer Knowledge Survey for Elders, was administered to participants in their native language (eight different language groups). Distribution was through community workers and public healthy nurses who worked with older adults. MAIN RESEARCH VARIABLES: Knowledge about cancer, language group, length of time living in the country. FINDINGS: The highest number of correct responses (87%) was for the item "Can some cancers be cured if they are discovered early?" The highest number of wrong answers (67%) was for the item "Can a bump or bruise to the body cause cancer?" Sixty-six percent did not consider age as a risk factor. For all but three items, the proportion of English language to non-English language individuals with correct answers was significantly different individuals whose native language was not English were less knowledgeable about cancer. IMPLICATIONS FOR NURSING PRACTICE: This study offers a basis for a large multicultural survey. Should the observations be confirmed in a larger sample, implications exist for public education about the risk factors and signs and symptoms of cancer, especially with individuals for whom English is not a native language. CONCLUSIONS: In this sample, specific areas were identified in which knowledge about cancer was lacking. In particular, the increased risk of cancer with advancing age was not recognized by a significant portion of study participants. PMID- 9399275 TI - Patients' knowledge of and attitudes toward the management of cancer pain. AB - PURPOSE/OBJECTIVES: To examine patients' knowledge of and attitudes toward the management of cancer pain and to identify, from the patients' perspectives, factors contributing to effective and ineffective pain relief. DESIGN: Descriptive, correlational. SETTING: Ambulatory care oncology facility in Canada. SAMPLE: Convenience sample of 42 patients receiving oral pain medication for chronic cancer-related pain. METHODS: Participants completed a modified version of the Patient Pain Questionnaire and a demographic questionnaire and responded to two open-ended questions. MAIN RESEARCH VARIABLES: Patients' knowledge of and attitudes toward cancer pain management and their perceptions of factors contributing to effective and ineffective pain relief. FINDINGS: Many patients locked knowledge of the principles involved in effective cancer pain management and had unrealistic concerns about taking pain medications. Significant negative relationships were found between pain intensity ratings and factors such as patients' knowledge of pain management, their level of satisfaction with pain relief, and their perception of the goal of pain management. Patients identified a number of impediments to effective pain relief, including concerns about addiction and various side effects to pain medications. CONCLUSIONS: Many patients have inadequate knowledge about the management of cancers pain and have unrealistic concerns about taking pain medications, both of which have been identified in the literature as barriers to effective cancer pain management. IMPLICATIONS FOR NURSING PRACTICE: A need exists for patient education that addresses patients' misconceptions and concerns about using pain medications and the principles involved in effective cancer pain management. PMID- 9399274 TI - The effectiveness of the breast self-examination facilitation shield. AB - PURPOSE/OBJECTIVES: To evaluate the degree to which the recently developed breast self-examination facilitation shield (i.e., breast shield) is effective in helping women detect breast lumps during breast self-examination (BSE). DESIGN: Women were randomly assigned to examine silicone lump breast models while using the breast shield and while not using the breast shield. SETTING: A university medical center school of nursing in a mid-Atlantic state. SAMPLE: 52 English speaking women, predominantly healthcare professionals, ages 18 through 67, who had no physical limitations that might impede lump detection using their fingers. MAIN RESEARCH VARIABLES: Lumps correctly detected, lumps falsely detected, and time needed to complete the examination while using or not using the breast shield during examination. FINDINGS: Women correctly identified more than half of the lamps in models, regardless of whether or not the breast shield was used. False lump detection was not increased while using the breast shield. Time taken to perform breast examination increased, often significantly (p < or = 0.05), while using the breast shield. CONCLUSIONS: Lump detection using the breast shield was similar to that of not using the breast shield. Use of the breast shield did not increase false positive reports and was associated with increased examination time. IMPLICATIONS FOR NURSING PRACTICE: The breast shield provides or useful complement to teaching BSE. While guiding a woman's fingers over her breast following the exam pattern, the nurse can indicate and teach about underlying breast structures and can provide an individualized, graphic guide to monthly BSE. PMID- 9399276 TI - The effects of infusion rate on platelet outcomes and patient responses in children with cancer: an in vitro and in vivo study. AB - PURPOSE/OBJECTIVES: To determine the influence of infusion rate on quality of transfused platelets and patients' physical and subjective responses. DESIGN: Linked in vitro and in vivo studies with repeated measures and crossover designs, respectively. SETTING: A 52-bed pediatric cancer center in the midsouthern United States. SAMPLE: In vitro: 12 randomly selected platelet units. in vivo: convenience sample of 26 children, ages 3-20 years, with cancer and thrombocytopenia requiring platelet transfusion. METHODS: Four infusion rates studied in vitro: two infusion rates studied in vivo. MAIN RESEARCH VARIABLES: Platelet count, morphology score, corrected count increment, and patients' physical and subjective responses. FINDINGS: No clinically significant differences were found in outcomes across the four rates in vitro. Similarly, no statistically significant differences were found between the two in vivo rates on objective or subjective outcomes. CONCLUSIONS: In this setting, the more rapid infusion rate is clinically preferable because it does not negatively affect platelet recovery or patient outcomes and it decreases the infusion time by half. IMPLICATIONS FOR NURSING PRACTICE: Findings provided a basis for altering platelet infusion rates at the study setting. Benefits of the faster infusion rate include more rapid correction of thrombocytopenia, decreased time that outpatients must remain in the clinic for transfusion. Increased time available for other parenteral infusions in the impatient setting, and substantial savings in nursing time and associated costs. PMID- 9399277 TI - A patient-education tool for patient-controlled analgesia. AB - PURPOSE/OBJECTIVES: To develop a pamphlet for educating patients about patient controlled analgesia (PCA). DATA SOURCES: Journal articles and pump manufacturers' materials. DATA SYNTHESIS: This pamphlet defines PCA and describes PCA pump operation, pain assessment, medication side effects, and safety considerations. A numerical pain-assessment tool also is included. CONCLUSIONS: This pamphlet has been helpful in assisting patients to use PCA pumps effectively. IMPLICATIONS FOR NURSING PRACTICE: Nurses can use this tool to educate patients requiring PCA therapy for pain management. PMID- 9399278 TI - Repetitive practice of a single joint movement for enhancing elbow function in hemiparetic patients. AB - The primary goal of this study was to assess whether repetitive practice of flexion-extension movements of the affected elbow in hemiparetic patients enhances performance and to compare the effects of this practice mode to the effects of the physical therapy variable exercise program which is routinely applied during sessions. Subjects were 27 poststroke hemiparetic patients, residents of a rehabilitation institute, divided into an experimental (n = 15) and a control group (n = 12). The former were treated with 800 repeated elbow movements in a maximal predetermined amplitude of 80 degrees, provided in 8 equal sessions every other day. The latter received 10 min. of conventional physical therapy for the paretic upper extremity at similar time intervals. Pre- and posttreatment assessments included the bilateral measurements of kinematic variables and activation latencies of the biceps and triceps brachi muscles as well as motor and functional tests. For all criterion variables, the findings pointed to comparable improvement in both groups. It was concluded that repetitive elbow movements had no unique training effect on the kinematics of movement and on activation latencies of the primary muscles controlling elbow function in hemiparetic patients. Further, transfer of the effects of training to execution of movements towards and from the mouth was also comparable in both groups, pointing again to there being no particular advantage in using repetitive movements as a training mode for enhancement of elbow function in hemiparetic patients. PMID- 9399279 TI - Brain hemisphere dominance and personality. PMID- 9399280 TI - A facilitator in self-reported perception of physical symptoms: the role of contingency between physical symptom and aversive event. AB - Perceptions of physical symptoms are influenced not only by the passive process of physiological stimuli but also by psychological factors such as the contingency between a physical symptom and an aversive event which we examined here in two experiments. 18 subjects in Exp. 1 and 14 subjects in Exp. 2 performed motor tasks. In the Physical condition, an aversive event was contingent on the physical symptom of 'racing heart' in Exp. 1, and on the symptom of 'overstraining shoulder muscles' in Exp. 2. In the Motor condition, an aversive event was contingent on the response of the 'disordered pace of motor tasks' in both experiments. Self-reported scores on attention to and perception of the physical symptoms under the Physical condition were higher than those under the Motor condition. However, there were no differences in the actual physical responses between the two conditions. These results suggest that a contingency between a physical symptom and an aversive event facilitates attention to and perception of the physical symptom. PMID- 9399281 TI - Loneliness and health-related variables in young adults. AB - In classrooms, 69 young adults responded to the Revised UCLA Loneliness Scale, the Symptom Pattern Scale, and the General Health Rating Index, a measure of perceived health status. A statistically significant positive correlation of .21 was found between scores for loneliness and ratings for symptom patterns. A statistically inverse correlation of -.35 was found between scores for loneliness and ratings for perceived health status. These findings replicated those found earlier with adolescents. PMID- 9399282 TI - The Multidimensional Dream Inventory: preliminary evidence for validity and reliability. AB - The present study focused on the development of the Multidimensional Dream Inventory, an individual difference measure of dimensions of dreams. Items were administered to 151 college students. Consistent with expectations, results of an exploratory factor analysis of intercorrelations among items indicated a four factor solution was appropriate. As a result, four dream-relevant scales were constructed, viz, Dream Importance, Dream Vividness, Dream Usefulness, and Dream Recall. In addition, these scales showed good internal consistency for research. Implications and uses for the Multidimensional Dream Inventory were discussed. PMID- 9399283 TI - Speech and language characteristics of children with strokes due to sickle cell disease. AB - The receptive and expressive language skills of 10 children with strokes due to sickle cell disease were significantly poorer than those of their matched controls. The children with strokes had greatest difficulty in following oral directions and formulating sentences. PMID- 9399284 TI - Correlations for social support with depression in the chronic postsroke period. AB - This study examined correlations of social support with rated mood states, including depression, for 47 patients with cerebrovascular disease during the chronic poststroke period. After the Structured Clinical Interview for DSM-III-R, four psychological measures, the Zung Self-depression Scale, the Hamilton Depression Scale, Profile of Mood States, and Social Support Scale, were administered. The patients with cerebrovascular disease exhibited significantly more psychiatric disorders, including depression, and had poorer social support than healthy controls. The severity of depression was significantly related to poor social support and particularly to the presence of social support rather than just the perception of poor social support. Depressed patients may also rate their support as poor because they are depressed. For some patients with cerebrovascular disease during the chronic poststroke period, depression may be related to low social support. PMID- 9399285 TI - Effects of instruction and practice on the length-reproduction task using the Muller-Lyer figure. AB - Two experiments were conducted to investigate the effects of instruction and practice on the length-reproduction task using the Muller-Lyer figure. The task consisted of reproducing visually presented lines by positioning the arm. In Exp. 1, 20 subjects were asked to move a stylus exactly the same length as the line presented in the illusory figure. Instructed subjects were told to ignore the visual illusion, while uninstructed subjects were told nothing. No differences in groups' constant errors were observed; however, subjects in both groups reproduced longer or shorter lines in accordance with the visual illusion. In Exp. 2, we investigated the effect of visual illusion in relation to learned movement. After practicing a length reproduction task 200 times, subjects performed a test session in which illusory figures were among the presented lines. Subjects were instructed to ignore any visual illusions, but this again had no effect on the reproduced length, that is, longer or shorter lines were reproduced in accord with the illusion. The findings show that visual illusion still had an effect after subjects were instructed to ignore it or had practiced the task extensively. PMID- 9399286 TI - Students' tactics of resistance and teachers' stress. AB - In a study of 134 college teachers, teachers' self-reported stress scores were significantly and positively related with their perceptions of students' use of reluctant compliance and deception tactics in resistance to on-task learning. PMID- 9399287 TI - Worth Index. AB - A measure of the perception of the source of one's worth was developed. The Worth Index focussed on whether people think that the value or worth of their lives is something which they must earn or is something that is innate and therefore cannot be earned. Estimates of validity and reliability are reported. PMID- 9399288 TI - Effects of virtual reality-enhanced exercise equipment on adherence and exercise induced feeling states. AB - A field study was conducted to test the effectiveness of virtual reality-enhanced cardiovascular exercise equipment for increasing adherence and attendance in a mixed-sex adult sample. Attendance was significantly higher in the virtual reality-enhanced condition than in the conditions without virtual reality over the 14-wk. period. Adherence was also highest (83.33%) in the virtual-reality bicycle group. Postexercise feelings of positive engagement, revitalization, tranquility, and physical exhaustion, as measured by the Exercise-induced Feeling Inventory, did not differ among groups. Contrary to previous findings, Self motivation Inventory scores were not associated with either attendance or adherence. While findings suggest that virtual-reality features may promote exercise adherence or attendance, it is not yet known what psychological variables they affect. Implications were drawn regarding the practical possibilities for exercise promotion. PMID- 9399289 TI - Human ethology: age and sex differences in mall walking. AB - Well-controlled experimental research has examined the biomechanical aspects of walking in homo sapiens on a track. The research reported here also examined cadence, velocity, and stride length for estimated ages ranging from 15 to over 55 years but in a shopping mall. Women at all ages walked faster than men in the mall setting which was opposite to what was found in the track research. Apparently context may influence how fast people walk. Hunter-gatherer differences could explain these results. PMID- 9399290 TI - Percept-genesis of the mother-child separation theme among panic and asthma patients. AB - A stimulus portraying a mother figure who is leaving a baby alone on the floor (Separation Theme) was presented tachistoscopically at increasing exposure times, according to the method of the Defense Mechanism Test, to three sex-matched groups of 31 normal subjects, 31 patients with bronchial asthma, and 31 patients with Panic Disorder and Agoraphobia. The frequency of several codings was significantly higher in both clinical groups compared with normal controls. Asthma patients were characterized by reports of the child seen as a statue and of contact or fusion between mother and child. Agoraphobic patients employed different strategies, centered on the mother rather than on the child and mainly represented by the denial of mother's action, e.g., she is not leaving, she is entering. The findings support the hypothesis of a difference in defensive organization between neurotic and psychosomatic patients. PMID- 9399291 TI - Criteria used to judge obese persons in the workplace. AB - Researchers have speculated that employers are less likely to hire obese persons for more publicly visible jobs, although this hypothesis remains untested. In the present study, 54 undergraduate students rated 40 jobs on several items, including the likelihood they would hire an obese person for each job. Multidimensional scaling showed a one-dimensional solution, labeled as physical activity, with participants less likely to hire obese persons for more active jobs. For hiring likelihood ratings for jobs at either end of the dimension appear to be most similar for men and individuals with more positive attitudes toward obese persons versus women and individuals with more negative attitudes toward obese persons. Implications for both theory and practice are discussed. PMID- 9399292 TI - Maintenance of exercise behavior for individuals at risk for cardiovascular disease. AB - The purpose of the study was to examine psychological factors associated with maintenance of exercise behavior in a population of middle-aged individuals with elevated risk factors for cardiovascular disease. 191 males and 17 females took part in a one-year diet and/or exercise intervention during 1990-1991. Four years later questionnaires were sent out to the 200 former participants who were still available for contact. 67.9% of those who answered (n = 140) were categorized as exercisers, and 30.7% were categorized as nonexercisers. The majority of the exercisers had exercised at least one and a half years. A chi-squared analysis showed that whether the individuals were exercising or not at present was independent of whether they had exercised or not during the intervention study. Discriminant analyses were used to determine how well physical self-perceptions at different times would categorize exercisers and nonexercisers. Current physical self-perceptions categorized the Active Exercisers (86.9%) and the Nonexercisers (63.3%) the best (in total 79.1% correct classifications). Neither change in physical self-perceptions during the intervention not change in physical self-perceptions from the end of the intervention until four years later, classified the exercise behavior as well. Three social cognitive models, The Self-perception model, The Health Belief model, and The Self-efficacy model, were investigated as discriminators between Active Exercisers and Nonexercisers. Active Exercisers were classified better than Nonexercisers, and current physical self-perceptions showed the highest percentage of total correct classifications. The proposed models were also analyzed as predictors of the variance in self rated Motivation for Exercise. Outcome Expectations, Compliance Self-efficacy, Perceived Fitness, and Exercise Mastery explained 45% of the variance in self rated Motivation for Exercise. PMID- 9399293 TI - Creative synthesis of visual images is not associated with individual differences. AB - 50 undergraduates were tested on a mental synthesis task aimed at producing creative visual patterns and were administered three questionnaires measuring imagery vividness and control. Analysis did not support a relationship between scores on visual synthesis and imagery and showed that neither kind of score was influenced by sex and studies attended. PMID- 9399294 TI - Of time and preference: temporal stability of environmental preferences. AB - Time is a central issue in discussions about art and in Berlyne's aesthetic theory. This article reports on the temporal stabilities of preferences for a novel and controversial building at three times after construction (2 years, 18 years, 23 years), and public preferences for 20 ordinary and noncontroversial buildings at three times over nine years. In all there were 5543 respondents. Analyses suggested that the initial response to the novel building was stable over the next 23 years, and the public responses for the 20 nonnovel buildings were stable over nine years. Implications for research are discussed. PMID- 9399295 TI - Fantasy and reality distinction of congenitally blind children. AB - 40 congenitally blind and 40 sighted children were tested for fantasy-reality distinctions of real and imagined objects and the development of concepts of darkness, hidden objects, space, dreams, emotions, facial expressions, size, and height. Analysis indicated that the blind children distinguished between contents of fantasy and reality, although they were less sure about the reality status of the objects. The sighted group gave more reality responses than the blind group for the concepts of dreams and hidden objects, but the remaining concepts were somewhat the same. Cognitive development explained in terms of theory formation may not explain the development of young blind children completely. Their knowledge that contents of fantasy are not real may be obtained through interpersonal experiences that are publicly shared. PMID- 9399296 TI - Development of a short test for accident proneness. AB - In developing the Accident Proneness Prediction Test to measure motor control skills related to safety aptitudes subjects were instructed to draw on a sheet of paper as many circles as possible within a limited amount of time. The responses were scored in terms of the speed and quality (irregularities) of the produced circles. Although 103 accident-prone drivers produced more circles than the 178 good drivers, the quality of performance of the accident-prone drivers was poorer than that of the good drivers. Based on this validity study, we proposed these predictive criteria for further testing of accident proneness. PMID- 9399297 TI - The Fitness Facility Membership Questionnaire: a measure of reasons for joining. AB - This study describes the development of the Fitness Facility Membership Questionnaire of 43 items, designed to identify reasons for joining a fitness facility. Items were generated from responses to an open-ended questionnaire. A 50-item version of the questionnaire was completed by 152 members of five community-based fitness facilities. Principal components analysis with varimax rotation yielded 8 factors, accounting for 63.8% of the variance. The factors were labeled Socialization, Aquatic-related Facilities, Extrinsic Motivation, Recreational Facilities, Intrinsic Motivation, Resistance Equipment, Aerobic Equipment, and Amenities. The internal consistency of the eight factors was acceptable with Cronbach coefficients alpha ranging from .72 to .89. Discriminant analysis of responses is also presented. PMID- 9399298 TI - Reproductive status of mothers affects sex-biased parental investment in humans. AB - An analysis of the sexual differences in birth weights of 381 children showed that boys are heavier than girls for multiparous mothers but not for primiparous ones. The results support the current hypotheses that predict sex biases in parental investment, with higher costs of producing male offspring in some mammals. PMID- 9399299 TI - Graphologists' assessment of extraversion compared with assessment by means of a psychological test. AB - The study investigated whether graphologists can infer extraversion from handwriting correctly. On the basis of three personality questionnaires, three persons (targets) were classified as extraverted and three as introverted. Ten graphologists independently analysed the handwriting of the targets and classified them as extraverted or introverted. Of the 60 (10 graphologists for 6 targets) classifications 58 were correct, which shows the graphologists assessed the classification of extraversion from handwriting. Graphologists agreed substantially on which characteristics of the handwriting were indicative for classification as extraversion or introversion. In each handwriting sample, however, both extraverted and introverted characteristics were present. Eventual classification may be based on the relative frequency of the two kinds of characteristics. Comparative studies like this one indicate that in research one should consider whether graphologists and psychologists share the same notion of extraversion. PMID- 9399300 TI - Assessing lumbar stabilization from point-light and normal video displays of manual lifting. AB - Routinely, physical therapists use visual observation to assess qualitatively a patient's performance. The literature, however, indicates that assessments of gait and lumbar stabilization from visual observation are at best only moderately reliable. Point-light video displays have been used to study the visual perception of human motion. The present purpose was to examine the reliability of assessments made by a physical therapist when viewing point light and normal video displays of subjects performing a lifting task. Three physical therapists assessed lumbar stabilization by viewing normal and point-light displays of 25 subjects who lifted an 8-lb. milkcrate from floor to waist height. Greater agreement of the therapists' ratings of lumbar stabilization was achieved on assessments made from point-light displays than on those made from normal displays. This finding suggests that the use of point-light displays may improve the reliability of qualitative assessments of performance on motor tasks. PMID- 9399301 TI - Paranormal beliefs and personality scores of high school students. PMID- 9399302 TI - Alexithymic characteristics in responses to the Synthetic House-Tree-Person (HTP) Drawing Test. AB - This study examined the association of certain complex personality traits assessed by the Synthetic House-Tree-Person Drawing Test and alexithymic characteristics assessed by the 20-item Toronto Alexithymia Scale for a sample of 589 Japanese college students. Alexithymic students who scored over 61 points on the Toronto Alexithymia Scale-20 exhibited two characteristics relative to the test: poor relationships between figures and additional written explanations. These two characteristics projected on the Synthetic House-Tree-Person Drawing Test may be related to alexithymic characteristics and related factors. PMID- 9399303 TI - Perception of well-being among older persons in nonmetropolitan America. AB - This study examined the relationships of some selected sociodemographic variables to perception of well-being by elderly individuals living in nonmetropolitan areas in the United States. Data used were from the National Opinion Research Center's (NORC) General Social Surveys for the ten years between 1982-1991. Seven sociodemographic variables and six attitudinal variables were considered independent variables, and perception of well-being was treated as the dependent variable. Analysis of variance and multiple regression analysis indicated marital status, education, financial status, and religious attendance were significantly related to perception of well-being and five attitudinal variables increased the total variance accounted for in perceived well-being. PMID- 9399304 TI - Spatial representation in face drawing and block design by nine groups from hunter-gatherers to literates. AB - A rank-order correlation was performed for nine cultural groups ranging from preliterate hunter-gatherers to literate medium-city dwellers. Two spatial tests of intrapattern spatial relations were used, the Draw-A-Person-With-Fade-In-Front test and the Kohs Block Design, a test of constructive praxia. In contrast to traditional "Western" evaluations, credit was given for the preservation of the essential intrapattern shapes even when exact spatial relations among these shapes was incorrect. Such "errors" were labelled "neolithic face" patterns and "nonrandom errors," respectively. Analysis suggested that the neglected intrapattern (in contrast to interobject) spatial relational skills emerge with literacy but is not yet actualized in preliterates whose survival requires quick fight or flight response upon prompt, albeit gross, assessment of salient shapes of prey or predators (human or animals). The positive Spearman rank-order correlation of absent or low literacy skills with the percent of "neolithic face" drawings was .95 and with the "nonrandom" block designs .67. Suggestions were developed for assessing certain unusual "ecological" present situations or certain brain dysfunctions. PMID- 9399305 TI - Aesthetic preferences for combinations of color and music. AB - 135 university undergraduates heard 12 preludes from J. S. Bach's Well-tempered Clavier (Vol. 1) while viewing alternating red, yellow, green, and blue colored lights. Their task was to rank-order the lights according to how well they "matched" the music. Preferences for combinations of color and music differed depending on whether the music was in a major or a minor key. The present findings along with those of some earlier studies suggest that aesthetic experience may be heightened when colors are seen that match the mental images music evokes. PMID- 9399306 TI - Comparisons of teaching presentation and development of content: implications for effectiveness of teaching. AB - The purpose of this study was to describe the choices of content development and task presentations of four teachers with varying experience and the subsequent achievement in skills test of their 125 junior high school students over the course of an 11-day volleyball unit. The results supported the importance of teachers presenting clear, detailed, and game-oriented progressions of content and tasks to improve students' learning. PMID- 9399307 TI - A modified step test based on a function of subjects' stature. AB - A number of submaximal step tests have been developed to predict maximal aerobic capacity. Because step height may influence biomechanical efficiency and heart rate, step tests based on subjects' stature may more accurately predict maximal aerobic capacity. Eighteen women performed the Queens College step test and a modified Queens College step test. The modified step test was performed with the height of the bench set even with the height of the foot at a knee angle of 90 degrees. Analysis of the data indicated a lower recovery heart rate following this test (p < .05). Further, correlations between maximal aerobic capacity and recovery heart rate for both tests were moderate (r = -.80 and -.75, respectively). Our results suggest that step tests based on subjects' stature do not more accurately predict aerobic capacity than those using a standardized bench height. PMID- 9399308 TI - The Health Student Academic Locus of Control Scale. AB - The Health Student Academic Locus of Control Scale is a 20-item context-specific scale, developed to measure Internal and External control beliefs of students in courses allied to medicine. Psychometric properties are acceptable (N = 164) so the scale can be used to measure control beliefs in a longitudinal study. PMID- 9399309 TI - Computer-aided cognitive rehabilitation: possible application to the attentional deficit of schizophrenia, a report of negative results. AB - The cognitive deficits associated with schizophrenia commonly include impairment in attention, which may contribute to difficulties with learning, memory, and executive function. This study evaluated the effectiveness of computer-aided training of attentional skills in schizophrenia. Two groups of schizophrenic subjects (9 men and 1 woman) were matched for age, estimated premorbid IQ, and positive and negative symptom scores. Both groups were assessed using a battery of attentional tests. Subjects then received either six 1-hr. computer-aided cognitive rehabilitation sessions (experimental condition) or six sessions of graphics-based computer games (control condition). Both groups were reassessed with attentional measures. There was significant improvement on only one test, a letter-cancellation task. This improvement was evident in both groups suggesting that this was a practise effect. Apart from the letter-cancellation test, subjects undertaking the computer-aided rehabilitation treatment did not show significant improvement on any attentional tasks. PMID- 9399310 TI - Morphed images of basic emotional expressions: ratings on Russell's bipolar field. AB - An object may be gradually changed into another object by a technique called "morphing" as in the current study. Although in some studies, such as by Coren and Russell, subjects were asked to rate psychologically synthesized facial images seen in a stereoscope, there do not seem to be any studies in which facial images were physically synthesized as morphed images. Our question then was how subjects would rate them. Multidimensional scaling indicated that ratings of facial images, physically synthesized by morphing, showed essentially the same configuration as those of psychologically synthesized faces and that our results conformed to the well-known dimensions of emotional space. In contrast, ratings of fear might not conform to the dimensional emotional space, suggesting cultural differences in emotion between Japan and the United States. PMID- 9399311 TI - Speed and accuracy in the learning of a complex motor skill. AB - The effect of emphasizing speed or accuracy on the learning of a high speed-high accuracy skill, the fastpitch softball pitch was investigated. 26 10- and 11-yr. old girls were randomly assigned to two groups receiving feedback on speed of throwing or accuracy of throwing during a 6-wk. training. Measurements of speed and accuracy were made and recorded on all participants at each practice session and a videotape of their pitching technique was also made at each session. Data were subjected to 2 x 3 (2 groups by 3 testing times) repeated-measures analyses of variance. The speed group threw faster and with better technique during the study and was able to maintain speed and accuracy in the reversed test condition. PMID- 9399312 TI - Confirmatory factor analysis of the French translation of the 20-item Toronto Alexithymia Scale. PMID- 9399313 TI - Influence of physical exercise on simple reaction time: effect of physical fitness. AB - The influence of physical fitness and energy expenditure on a simple reaction time task performed during exercise was investigated. Two groups of 10 subjects were used, one was composed of trained middle-distance runners and one of students who had no regular physical training. The subjects performed a simple reaction time task while pedalling on a cycloergometer at different relative power output corresponding to 20, 40, 60, and 80% of their own maximal aerobic power and immediately after exercise. During exercise, the results showed a decrease in cognitive performance for both groups whereas no significant effect was found after exercise. A significant effect of physical fitness on simple reaction time was noted during exercise. The data are interpreted in terms of optimization of performance focusing particularly on the relations between energy cost of the physical task and attentional demand. PMID- 9399314 TI - Spatial visualization: running visualization for empty squares. AB - A technique is reported in which an oral description of empty squares had to be visualized. A tone signaled completion, and a response was made whether the last square had touched a previous square. The chain lengths varied from 10 to 19 squares at closure. Local closure sizes consisted of 4 to 10 squares. Closures up to 6 squares produced the least errors. There were biases in response as a function of response type and chain length. The technique could be useful for monitoring continuous spatial visualization. PMID- 9399315 TI - The aetiology and long-term effects of injuries due to bicycle accidents in persons aged fifty years and older. AB - This retrospective study concerns the aetiology and psychological long-term effects of injuries due to bicycle accidents in 329 patients 50 years and older who attended the Emergency Unit of the University Hospital at Groningen during the period 1990 through 1992. Long-term effects were assessed three years after hospital discharge. The one-sided bicycle accident (with no other traffic involved) was the major (63.2%) cause, mostly due to loss of balance or to a foot slipping from the pedal. The main category of the second major cause was collision with other traffic. Of the bicycle accidents 66% occurred within 15 minutes after departure; 80% of the accidents happened in good weather conditions and daylight, and 7.6% of the patients had taken tranquilizers before biking. The majority of the injuries were observed at the upper extremities (28.8%) and head or face (25.8%). The percentage of clinically treated patients increased across ages from 25% in the 50- to 54-yr.-old category to 45% in the category 75-yr. and older. Three years after the incident, long-term psychological effects were still observed in 29% of the patients. PMID- 9399316 TI - Order effect for two measures of hypnotizability. PMID- 9399317 TI - Depth perception in simple line drawings. AB - Three-dimensional interpretation of simple line drawings, composed of two triangles with a common side, was studied through the quantitative measurement of perceived orientation of the surface indicated by a stimulus figure. In a single triangle, depth perception is ambiguous and is not stable even if perceived. In two triangles with a common side, however, depth is stably perceived. Depth effect, defined as the magnitude of the angle formed by the two perceived surfaces, increased linearly as the magnitude of an angle at a vertex facing the common side became larger. The depth effect did not vary significantly for the change of a triangular from when the magnitude of the angle at the vertex facing the common side was constant. These results suggest that the depth effect changes systematically with variation in the triangle's form. PMID- 9399318 TI - Spring peak in suicides. PMID- 9399319 TI - Magnitude estimation scaling of the loudness of a wide range of auditory stimuli. AB - The study of the perception of loudness lends itself well to the psychophysical scaling technique of magnitude estimation. This study was designed to extend the range of auditory stimuli used to study the magnitude estimation scaling of loudness. The five stimuli chosen were a 1000-Hz pure tone, narrow band noise (700-1300 Hz band width), broad band noise (100-10,000 Hz band width), rock music, and babble speech, i.e., speech in which meaning is not discernible because several individuals are talking at once. Subjects were 30 normal young women (M = 19 yr.). During the auditory magnitude-estimation task for each of the five stimuli, a subject was instructed to assign numbers to stimulus presented in a randomly ordered series of nine sensation levels (10, 20, 30, 40, 50, 60, 70, 80, and 90 dB SL). Multivariate analysis of variance for repeated measures indicated there were no significant differences in the numerical responses of the subjects for the five stimuli. A possible explanation for these results is the presence of an underlying stabilizing factor (internal scaling mechanism) that allows adults to scale loudness consistently irrespective of the type of auditory stimulus. PMID- 9399320 TI - Speech delay in seven siblings with unusual sound preferences. AB - By the age of 8 years, children who are developing normally show almost adult speech skills. Children with serious phonological disorders, however, may exhibit significant differences in development well beyond the age of 8 years with little or no improvement in speech if therapy is not provided. This is a descriptive study of seven siblings, ranging in age from 6 to 14 years of age who had never attended school or received speech therapy until these ages. All of the children exhibited moderate to severe speech disorder with no evidence of predisposing genetic factors, hearing loss, physical abuse, or prenatal drug exposure. These cases, which would obviously be impossible to duplicate in a controlled study, provide strong support for the efficacy of speech therapy. Children with serious speech delays will not improve appreciably without direct intervention. PMID- 9399321 TI - Relationship between body image and percent body fat among male and female college students enrolled in an introductory 14-week weight-training course. AB - Students (39 men and 27 women) from a southern university, who were enrolled in a 14-wk. introductory weight-training course, were administered a 20-item body image questionnaire and subsequently underwent skinfold measurements to assess percent body fat. Mean scores were correlated with percent body fat. For men, women, and both sexes combined correlations were significant and inverse (rs = .68, -.41, -.66, respectively). Body image as measured was inversely related to percent body fat among these college students. Researchers should examine how dietary and exercise-induced changes in adiposity (pre-post design) influence scores on body image. PMID- 9399322 TI - Cognitive correlates of complexity of children's drawings. AB - 240 children (60 each at ages 4, 6, 8, and 10 years) were administered Dennis' (1987) Five Drawing Tasks and five additional developmental tasks. Three hypotheses were tested: that object recognition and working memory would be related to increasing complexity, that both would load on separate factors, and that higher-order analyses would indicate an underlying second-order spatial factor. Analysis included very strong zero-order correlations with age. When age was partialed out, three first-order factors were obtained. Higher-order analyses yielded one second-order factor which appeared related to a general factor of spatial intelligence. PMID- 9399323 TI - The birthday blues. PMID- 9399324 TI - Possible interaction between vibration thresholds by sex and motor dominance in the index finger and big toe. AB - Two studies suggest a possible interaction among sex, motor dominance, and vibrotactile threshold for the great toe and index finger. In study 1 a forced choice procedure with the Vibratron II (Physitemp Instruments, Inc.) was used; a significant interaction between sex and foot dominance for vibratory threshold was noted with no main effects for the great toe. The greatest difference between men and women was on the nondominant side on the foot. Study 2 replicated Study 1 using the index finger as well as the great toe and used the Semmes. Weinstein monofilament test for a cross-modal comparison. A method of limits procedure was used to increase the generalizability of the data. A similar interaction was found between sex and motor dominance for the index finger but not the great toe. This was attributed to skewing of data for the toe. No effects were found for the Semmes-Weinstein test. Possible usefulness in detecting neuropathies is considered. Larger normative studies including variables such as age, height, and weight are required for generalizable conclusions. PMID- 9399325 TI - Affect identification bias demonstrated with individual chimeric faces. AB - The current study extends previous findings of a left visual-field bias in chimeric face tasks, by using a new procedure which incorporates chimeric stimuli depicting both positive and negative target affects and requires the identification of affect in individually presented faces. This new procedure is more representative of the types of judgements made in daily social interaction. Results with this new procedure are consistent with previous findings, indicating a significant left visual-field bias for both positive and negative affects in the majority of subjects. Handedness was significantly related to lateralization scores, with dextrals showing greater left visual-field biases than sinistrals. Among sinistrals, a left visual-field bias was noted only for happy chimera. PMID- 9399326 TI - A parental survey of speech, language, and physical development of infants and toddlers with sickle cell disease. AB - 16 parents of infants and toddlers (9 girls, 7 boys) with sickle cell disease reported normal speech and language development in their children but perceived slow physical development. Such perceptions may be related to the smaller physical size of children with this disease. PMID- 9399327 TI - Language of sport fans: sportugese revisited. AB - In 1959, Tannenbaum and Noah reported that sports writers and readers possessed a better understanding of sport terminology than nonreaders. The current investigation extended Tannenbaum and Noah's research using current sport terms. A positive relationship between understanding sport terminology, extent of team identification, strength of sport fandom, and self-proclaimed sport knowledge was hypothesized. Scores of 57 participants confirmed the predicted pattern. Discussion concerned research examining sport terminology. PMID- 9399328 TI - Aiming accuracy of the line of gaze and redesign of the gaze-pointing system. AB - The current experiment tested the aiming accuracy of the eye for targets at nine different positions and of two different sizes. The left eye was tracked to record the line of gaze with an infrared eye-monitoring device. For each subject, horizontal shift, vertical shift, standard deviations of the X and Y coordinates, and 95th percentile of the horizontal and vertical shifts from the nine target centers were calculated to indicate the precision, accuracy, and possible design limits of the gaze points. Analysis showed that the target's position had a strong effect on the vertical shift, standard deviations of the Y coordinate, and 95th percentile of the vertical shift. The effect of target size was not significant on any of the dependent measures. The research findings have important implications for the redesign of the gaze-pointing device. Visual targets appearing below the horizontal line of sight should be larger than those above. Based on the 95th percentile of the horizontal and vertical shifts, targets should be approximately 2.0 degrees wide and 2.4 degrees long above or coinciding with the horizontal line of gaze and 2.6 degrees wide and 3.9 degrees (cm) long below the horizontal line of gaze. PMID- 9399329 TI - Landmark enhancement and strategic processing: an evaluation of strategies for spatial navigation training. AB - Viewing a filmed route offers an alternative to more expensive and rigid methods of learning navigation skills. One advantage of film is the ability to enhance important landmarks or focus on particularly relevant information. The current research used a videotape of a spatial route to assess the usefulness of participants' interaction with landmarks and enhancement of landmarks for training spatial navigation. 48 participants were exposed to one of four videotaped route conditions: Landmark-enhanced, Question-based Interaction, Landmark-enhanced Plus Question-based Interaction, or Control (Nonenhanced without Question-based Interaction). Following the spatial navigation training with videotape, participants were asked to traverse the route in the actual building. Analyses of variance indicated that the Question-based Interaction group made significantly fewer wrong turns during traversal of the route than did the Control group. Also, enhancement of the landmarks did not significantly reduce wrong turns; in fact, it may have hindered the benefit of question-based interaction in reducing wrong turns. PMID- 9399330 TI - Perceptual defense and vigilance to perinatal stimuli. AB - The presentation times (milliseconds on a computer screen followed by a masking grid) required for the correct identification of tachistoscopically presented perinatal stimuli were compared for 30 pregnant women and 25 perimenopausal women. Analysis indicated a differential facilitation or inhibition of perception in logical relation to subjects' closeness to pregnancy or menopause: pregnant women are quicker to identify stimuli related to pregnancy or babies but slower to recognize pictures of a pregnant woman with her father or mother. This supports the validity of measurements based on the theory of perceptual defense or vigilance. PMID- 9399331 TI - The NMDA receptor competitive antagonist CPP modulates benzodiazepine tolerance and discontinuation. AB - Benzodiazepine discontinuation is characterized by a syndrome of increased activity and reduced seizure threshold that is similar to effects mediated by the glutamatergic system. To elucidate the involvement of the glutamatergic system in benzodiazepine tolerance and discontinuation, we administered lorazepam, the NMDA antagonist CPP, and the combination of these compounds either concomitantly or consecutively to mice via osmotic pumps and evaluated pentylenetetrazole-induced seizure threshold, open-field activity, and benzodiazepine receptor binding during and after chronic administration. Animals receiving lorazepam alone developed partial tolerance at 7 days and complete tolerance at 14 days to the anticonvulsant effects of lorazepam. This effect was partly attenuated by CPP coadministration with lorazepam. This combination produced only partial tolerance. A reduction in seizure threshold was observed 4 days after discontinuation of lorazepam alone. This effect was abolished by coadministration of CPP with lorazepam and by CPP administration during the withdrawal period. Benzodiazepine binding in most structures examined was significantly reduced at 14 days during chronic lorazepam administration (versus 1 day), and coadministration of CPP did not alter this decrement. After lorazepam discontinuation, binding was increased at 4 and 7 days versus chronically treated animals and versus vehicle within the cerebral cortex. This effect was abolished by coadministration of CPP as well as by CPP administration during the lorazepam withdrawal period. These data support the involvement of the glutamatergic system in benzodiazepine tolerance and discontinuation. PMID- 9399332 TI - Effects of phenytoin on the amygdala neurons in vitro. AB - The present study was aimed at elucidating the possible mechanisms underlying the anticonvulsant efficacy of phenytoin using intracellular recording techniques in the in vitro amygdalar slice preparation. Synaptic response mediated by the N methyl-D-aspartate (NMDA) receptor (EPSPNMDA) was isolated pharmacologically by application of a solution containing non-NMDA receptor antagonist 6-cyano-7 nitroquinoxaline-2,3-dione (10 mumol/l) and gamma-aminobutyric acidA receptor antagonist bicuculline (20 mumol/l). Phenytoin inhibits the amplitude of EPSPNMDA without affecting the postsynaptic depolarization induced by exogenous application of NMDA. In addition, phenytoin increases the magnitude of paired pulse facilitation which is consistent with a presynaptic mode of action. These results suggest that inhibition of transmitter release due to presynaptic blockade of Na+ and/or Ca2+ channels may account largely for the anticonvulsant efficacy of phenytoin. PMID- 9399333 TI - Metabolism of the tricyclic antidepressant amitriptyline by cDNA-expressed human cytochrome P450 enzymes. AB - The metabolism of amitriptyline was studied in vitro using cDNA-expressed human cytochrome P450 (CYP) enzymes 1A2, 3A4, 2C9, 2C19, 2D6 and 2E1. CYP 2C19 was the most important enzyme with regard to the demethylation of amitriptyline, the quantitatively most important metabolic pathway. CYP 1A2, 3A4, 2C9 and CYP 2D6 also participated in the demethylation of amitriptyline. CYP 2D6 was the sole enzyme mediating the hydroxylation of amitriptyline, and (E)-10-OH-amitriptyline was exclusively produced. CYP 2E1 did not metabolize amitriptyline. Concerning the quantitative relations, CYP 2C19 and 2D6 exhibited high affinities with Km values in the range of 5-13 mumol/l, whereas the affinities of 1A2, 3A4 and 2C9 were somewhat lower with Km values ranging from 74 to 92 mumol/l. CYP 2C19 displayed the highest reaction capacity per mole with Vmax equal to 475 mol h-1 (mol CYP)-1. The other enzymes had Vmax values in the range of 90-145 mol h-1 (mol CYP)-1. Allowing for the typical relative distribution of amounts of CYP enzymes in the liver, a simulation study suggested that, at therapeutic doses, on average about 60% of the metabolism depended on CYP 2C19. At toxic doses, CYP 2C19 is expected to be saturated, and CYP 3A4 may now play a dominant role in the metabolism. PMID- 9399334 TI - Inhibition of sympathetic outflow by the angiotensin II receptor antagonist, eprosartan, but not by losartan, valsartan or irbesartan: relationship to differences in prejunctional angiotensin II receptor blockade. AB - It is well established that angiotensin II can enhance sympathetic nervous system function by activating prejunctional angiotensin II type I (AT1) receptors located on sympathetic nerve terminals. Stimulation of these receptors enhances stimulus-evoked norepinephrine release, leading to increased activation of vascular alpha 1-adrenoceptors and consequently to enhanced vasoconstriction. In the present study, the effects of several chemically distinct nonpeptide angiotensin II receptor antagonists were evaluated on pressor responses evoked by activation of sympathetic outflow through spinal cord stimulation in the pithed rat. Stimulation of thoracolumbar sympathetic outflow in pithed rats produced frequency-dependent pressor responses. Infusion of sub-pressor doses of angiotensin II (40 ng/kg/min) shifted leftward the frequency-response curves for increases in blood pressure, indicating augmented sympathetic outflow. Furthermore, pressor responses resulting in spinal cord stimulation were inhibited by the peptide angiotensin II receptor antagonist, Sar1, Ile8 [angiotensin II] (10 micrograms/kg/min). These results confirm the existence of prejunctional angiotensin II receptors at the vascular neuroeffector junction that facilitate release of norepinephrine. The nonpeptide angiotensin II receptor antagonist, eprosartan (0.3 mg/kg i.v.), inhibited the pressor response induced by spinal cord stimulation in a manner similar to that observed with the peptide antagonist, Sar1, Ile8[angiotensin II]. In contrast, equivalent doses (0.3 mg/kg i.v.) of other nonpeptide angiotensin II receptor antagonists, such as losartan, valsartan, and irbesartan, had no effect on spinal cord stimulation of sympathetic outflow in the pithed rat. Although the mechanism by which eprosartan, but not the other nonpeptide angiotensin II receptor antagonists, inhibits sympathetic outflow in the pithed rat is unknown, one possibility is that eprosartan is a more effective antagonist of prejunctional angiotensin II receptors that augment neurotransmitter release. Because eprosartan is more effective in inhibiting sympathetic nervous system activity compared to other chemically distinct nonpeptide angiotensin II receptor antagonists, eprosartan may be more effective in lowering systolic blood pressure and in treating isolated systolic hypertension. PMID- 9399335 TI - Effect of trilinolein on superoxide dismutase activity and left ventricular pressure in isolated rat hearts subjected to hypoxia and normoxic perfusion. AB - Oxygen-derived free radicals have been implicated in the development of myocardial injury during hypoxia/reperfusion. Antioxidants can effectively inhibit the formation of free radicals and ameliorate the myocardial damage which may occur during hypoxia/reperfusion. Trilinolein is a triacylglycerol recently purified from the traditional Chinese medicinal plant Panax pseudo-ginseng. It has linoleic-acid residues as the only type of fatty acid residue in all three esterified positions of the triacyglycerol. It has been proposed that decreased endogenous superoxide dismutase (SOD) activity may contribute to free radical mediated reperfusion injury of the ischemic myocardium. In the present study, when isolated rat hearts were subjected to hypoxia for 10, 30, 60 and 90 min without normoxic perfusion, a significant decrease in Mn-SOD activity was shown throughout the period of hypoxia, whereas the Cu.Zn-SOD activity was increased at 10 and 30 min but was not different from the baseline at 60 and 90 min of hypoxia. In rat hearts pretreated with 10(-7) mol/l trilinolein and subjected to 60 min of hypoxia without normoxic perfusion, Cu.Zn-SOD was augmented compared with baseline and compared with hearts subjected to 60 min of hypoxia without trilinolein, whereas Mn-SOD activity was still reduced compared with baseline, although less so than after 60 min of hypoxia without trilinolein. Pretreatment with trilinolein was associated with better preservation of left ventricular function during hypoxia and more rapid return to recovery during normoxic perfusion. This myocardial protective effect may be related to an antioxidant effect through potentiation of SOD, particularly Cu.Zn-SOD during hypoxia. PMID- 9399336 TI - Lidocaine attenuates mechanical and metabolic derangements induced by palmitoyl-L carnitine in the isolated perfused rat heart. AB - The effect of lidocaine on the palmitoyl-L-carnitine (PALCAR)-induced mechanical and metabolic derangements was studied in Langendorff rat hearts, perfused aerobically at a constant flow rate and paced electrically. PALCAR (5 mumol/l) increased the left ventricular end-diastolic pressure, decreased the left ventricular developed pressure (i.e., mechanical dysfunction), and decreased the tissue levels of adenosine triphosphate and creatine phosphate (i.e., metabolic change). These mechanical and metabolic alterations induced by PALCAR were concentration-dependently attenuated by lidocaine (20, 50 or 100 mumol/l). Nevertheless, lidocaine (20, 50 or 100 mumol/l) did not affect the mechanical function and energy metabolism of the normal (PALCAR-untreated) heart. These results indicate that lidocaine has a cardioprotective action against the PALCAR induced mechanical and metabolic derangements. PMID- 9399337 TI - Circadian rhythm and drug delivery design. PMID- 9399338 TI - New 5-aminoacyl-5,10-dihydro-11H-dibenzo[b,e][1,4]diazepin-11-ones with antiarrhythmic activity. AB - A series of new 5-substituted tricyclic 5,10-dihydro-11H-dibenzo[b,e][1,4] diazepin-11-ones was identified as potential antiarrhythmic agents against bradyarrhythmias [1, 2]. The in vitro and in vivo interactions of the compounds with muscarinic receptors and the antiarrhythmic activity were examined. In receptor binding studies some derivatives showed a high affinity to the cardiac M2 receptor (Ki 10 nmol/l), an equal or smaller affinity to cortical M1 receptor and a lower affinity to the glandular M3 binding site. Functional experiments showed the derivatives as competitive antagonists with high affinity to the cardiac and smaller affinity to the intestinal muscarinic receptor. In vivo experiments correspond with the M2 selectivity. First the vagal or agonist induced bradycardia was inhibited in rats and guinea pigs while the McNA-343 induced increase of blood pressure, methacholine-induced bronchi and bladder constriction as well as the salivation were inhibited only at higher doses. In conscious cats the tachycardia was examined in comparison with pupillomotoricity. The effect duration and the therapeutical range were determined in comparison to the M2 selective blocking agent AF-DX116. The antiarrhythmic activity was examined compared to quinidine sulfate in CaCl2-arrhythmia of rats, in atrial fibrillation and atrial flutter in dogs according to Scherf [2] and in electric induced atrial fibrillation under vagal stimulation in cats. In the atrial arrhythmias the derivatives are clearly longer effective than quinidine sulfate. The antiischemic activity was examined in the two-stages coronary ligature in dogs according to Harris. The long-running regularization of ectopies (about 2 h after i.v. injection) occurred without decrease of the heart rate, an effect particularly convenient to therapy of bradycardic dysrhythmias. PMID- 9399339 TI - Enzymatic degradation of the triterpenoid saponin helianthoside 2. AB - Helianthoside 2 (1), the main bisdesmosidic saponin of Helianthus annuus L. was converted to the products 2, 3 and 4 by several, optimized enzymatic hydrolysis methods. Monosaccharide units of the acylglycosidic at C-28 of the sapogenin bonded, linear chain oligosaccharide were cleaved, but the branched trisaccharide at C-3 of the aglycone are stable under the conditions used. Compounds 2 and 3 are new triterpenoid glycosides, which were characterized as 28-O-alpha-L rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranoside (2) and 28-O-alpha-L arabinopyranoside (3) of 3-O-[alpha-L-rhamnopyranosyl-(1-->3)][beta-D xylopyranosyl-(1-->4)] beta-D-glucopyranoside of 3 beta,16 alpha-dihydroxy-olean 12-en-28-oic acid. PMID- 9399340 TI - Synthesis, structures and biological activity of some 4-amino-3-cinnoline carboxylic acid derivatives. Part 3: 1,3-Oxazino[5,4-c]cinnolines and pyrimido[5,4-c]cinnolines. AB - 6,7,8-Substituted 4-amino-3-cinnolinecarboxylic acids 1 were condensed with acid anhydrides to corresponding 1,3-oxazino[5,4-c]cinnolines 2. The reactions of 1,3 oxazino[5,4-c]cinnolines 2 with amines, hydrazine, hydroxylamine, alanine ethyl ester provided pyrimido[5,4-c]cinnolines 3, 4. Selected 3-substituted 2 methylpyrimido[5,4-c]cinnolin-4(3 H)-ones have been screened for CNS activity. PMID- 9399341 TI - Synthesis and antimicrobial testing of 4H-1,2,4-triazole, 1,2,4-triazolo[3,4 b][1,3,4]thiadiazole and 1,2,4-triazolo[3,4-b][1,3,4]thiadiazine derivatives of 1H-benzimidazole. AB - Three novel series of benzimidazole derivatives namely 6-substituted 3-[1-(2 alkyl-1 H-benzimidazolyl)methyl]-1,2,4-triazolo[3,4-b][1,3,4] thiadiazoles 5a-h, 6-substituted 3-[1-(2-alkyl-1H-benzimidazolyl)methyl]-7H-1,2,4 -triazolo[3,4-b] [1,3,4]thiadiazines 6a-j and 6-thioxo-3-[1-(2-alkyl-1H-benzimidazolyl)methyl]-5,6 dihydro-1,2,4 -triazolo[3,4-b][1,3,4]-thiadiazoles 7a, b have been prepared by cyclization of the key intermediate 1-[(4-amino-5-mercapto-4 H-1,2,4-triazol-3 yl)methyl]-2-alkyl-1 H-benzimidazoles 3a, b. Furthermore, 1-[(4-arylideneamino-5 mercapto- 4H-1,2,4-triazol-3-yl)-methyl]-2-alkyl- 1 H-benzimidazoles 4a-h have been prepared and some of them were cyclized to 6-substituted 3-[1-(2-alkyl-1H benzimidazolyl)methyl]-1,2,4-triazolo [3,4-b][1,3,4]thiadiazoles 5d, h using thionyl chloride. The prepared compounds were tested for antimicrobial activity in vitro; they showed moderate activity. PMID- 9399342 TI - Determination of fluoxetine in human plasma using reserved phase HPLC. AB - A rapid, simple, accurate method for the determination of fluoxetine in human plasma is presented. Liquid-liquid extraction of fluoxetine was carried out using diethyl ether. Chlorprothixene was applied as an internal standard. The samples were chromatographed on a LiChrosorb RP-18 (10 microns) column and the mobile phase was acetonitrile/phosphate buffer pH 2.70 (9:1). The detection was carried at 254 nm. A linear quantitative response curve was generated over a concentration range of 100-600 ng/ml. Overall extraction efficiency of the extraction procedure was found to be 86 to 91% with a correlation coefficient of 0.992. PMID- 9399343 TI - Phenytoin transfer across the in situ perfused rat term placenta. AB - Transfer of phenytoin (PHT) across the rat term placenta perfused in situ was investigated and compared with that of antipyrine (AP) as a marker of passive diffusion. PHT was shown to cross the placenta with similar kinetics as AP did. Both the first order transfer constant (ktr = 0.070 min-1) and the first order equilibration constant (keq = 0.027 min-1) of PHT were comparable to those of AP (ktr = 0.046 min-1, keq = 0.022 min-1). Similarly, there were significant differences between PHT and AP in the foeto-maternal concentration ratio at equilibrium (FMCReq = 1.01 and 1.09, respectively). The present data indicate that the transfer of PHT through the rat placenta is governed by the same principles as that of AP, i.e. by the mechanism of passive diffusion. Surprisingly, maternal plasma protein binding of PHT (60.5%) did not seem to influence either its rate of transfer or its eventual foeto-maternal concentration ratio. PMID- 9399344 TI - Heat and ionic limitations do not change the inhibition pattern of ribosomal peptidyltransferase by aminohexosyl-cytosine nucleoside antibiotics. AB - In a cell-free system derived from Escherichia coli, we investigated the inhibition of peptide bond formation by blasticidin S at 100 mM NH4+ and 5 degrees C or at 50 mM NH4+ and 25 degrees C. At both conditions, a transient phase of competitive inhibition is observed, followed by a mixed noncompetitive phase. The two phases of inhibition are compatible with a model in which a slow isomerization of the ribosome-drug complex occurs, as a result of ribosomal conformational changes. After this step, the mutually exclusive binding between acceptor substrate and antibiotic is converted to simultaneous binding. In comparison with a previous study carried out at 100 mM NH4+ and 25 degrees C, the present results demonstrate that the ribosomal conformational changes induced by blasticidin S can occur irrespectively of the reaction temperature and the ionic conditions. PMID- 9399345 TI - The hydrolysis of an alkylammonium salt series by rat liver microsomal esterase. PMID- 9399346 TI - Chemical composition and antimicrobial activity of Tanacetum parthenium essential oil. PMID- 9399347 TI - Prenatal sonographic findings in 187 fetuses with Down syndrome. AB - We determined the type and frequency of abnormal sonographic findings in 187 Down syndrome fetuses. Examinations were performed transvaginally or transabdominally between 9 and 28 weeks' gestation. Consecutive scans performed prior to knowledge of the fetal karyotype (n = 144) were analysed separately for one of the participating centres. In 93 fetuses (49.7 per cent), a total of 138 abnormalities were observed. The most commonly detected anomalies were cystic hygroma and increased nuchal fold thickness (30.5 per cent), hydrops (9.6 per cent), cardiac defects (7.5 per cent), pyelectasis or hydronephrosis (5.9 per cent), echogenic bowel (4.8 per cent), and a large variety of internal organ abnormalities (16.0 per cent) which are not typically associated with Down syndrome. Two anomalies or three anomalies in the same fetus were observed in 21 and 5 fetuses, respectively. No patterns of concurrent malformations were apparent among these fetuses. Sensitivity for Down syndrome detection by ultrasound scans performed without knowledge of the fetal karyotype was 24.1 and 42.6 per cent before 13 weeks and between 14 and 23 weeks, respectively. We conclude that structural abnormalities are frequently observed in Down syndrome fetuses, but many sonographic findings are not typically associated with this syndrome. PMID- 9399348 TI - Women's opinions and the implications of first- versus second-trimester screening for fetal Down's syndrome. AB - Two groups of pregnant women were questioned regarding their opinions on serum screening for Down's syndrome in the first trimester of pregnancy. One group comprised 83 women attending our antenatal clinic who were questioned at the time of the existing second-trimester screening test. Seventy-six per cent of those who participated in the second-trimester screening programme would have preferred the test to have been in the first trimester, mainly because of the easier termination of pregnancy and/or the earlier reassurance provided. The remaining 24 per cent could see no advantage in the earlier time frame. Of the 49 women who had declined second-trimester screening, only two would have participated in screening had it been in the first trimester. The other group comprised those women attending our antenatal diagnosis clinic who were considering chorionic villus sampling (CVS). Forty-four per cent of these women would have allowed serum screening in the first trimester to influence their decision as to whether to undergo definitive prenatal diagnostic testing. In general, those women who made use of second-trimester serum screening would also do so in the first trimester. Those who declined the existing screening programme would also decline first-trimester screening. Many women currently deciding to undergo CVS would allow a first-trimester screening test to influence their decision. PMID- 9399350 TI - Elevated hCG as an isolated finding during the second trimester biochemical screen: genetic, ultrasonic, and perinatal significance. AB - This study was undertaken in an attempt to determine the significance of elevated maternal serum human chorionic gonadotropin (MShCG), in the presence of an otherwise normal screen with respect to fetal malformations, chromosomal aberrations, and pregnancy outcome. Targeted ultrasound findings and perinatal outcome of 298 women in whom serum hCG was > or = 2.5 MOM and who were screen negative for Down syndrome (the study group) were compared with a control group of 229 women in whom serum hCG as well as the other parameters were within the normal range. Genetic amniocentesis was performed in 125 women from the study group. Ultrasonically detected malformations were significantly more frequent among the study group (12 vs. 1, P = 0.01). Pregnancy complications were similar in the two groups, with the exception of pre-eclampsia-toxaemia, which was significantly more frequent in the study group (5 vs. 0, P = 0.02). There was one case of an abnormal karyotype (47,XXY). Although genetic amniocentesis does not appear warranted, isolated elevated MShCG levels during the second trimester screening was associated with an increased risk of fetal anomalies detected by ultrasound and of toxaemia of pregnancy. PMID- 9399349 TI - Minimally-invasive early prenatal diagnosis using fluorescence in situ hybridization on samples from uterine lavage. AB - A two-phase study was undertaken to examine the efficiency of using transcervical cells (TCCs) collected by uterine lavage and fluorescence in situ hybridization (FISH) for early prenatal diagnosis of fetal chromosome aneuploidy. Uterine lavage was performed in 50 women scheduled for elective termination of pregnancy (TOP, n = 35) or chorionic villus sampling (CVS, n = 15) between 6 and 11 weeks of gestation. TCCS were dissociated by trypsin and collagenase, and interphase FISH was carried out for chromosomes X, Y, 13/21, and 18. The phase I study comprised 36 women. The FISH results were compared with the cytogenetic analysis from long-term culture of villus samples collected at TOP or CVS. Among the 36 samples, 15 had a normal male karyotype and 21 had a normal female karyotype. FISH on TCCs correctly identified 13 out of the 15 pregnancies with a male fetus. In phase II, uterine lavage was performed on 14 women. The samples were first tested for the presence of trophoblasts with an anti-trophoblast antibody, GB25, by immunohistochemical staining. Among 12 GB25-positive samples, the FISH results corresponded to the fetal karyotype. One of the GB25-positive samples had five signals for the chromosome 13/21 probe. The cytogenetic analysis confirmed that the fetus had a karyotype of 47, XX, +21. In the GB25-negative samples, FISH failed to identify one male pregnancy. Follow-up was carried out on 13 ongoing pregnancies and no maternal or fetal complications were discovered. This study demonstrates that fetal chromosome numeration can be carried out using FISH on uterine lavage samples in early pregnancy. However, a specific fetal cell marker, such as specific anti-trophoblast antibody, is necessary to avoid a false negative result. PMID- 9399351 TI - Prenatal diagnosis of 22q11 microdeletion. AB - Microdeletion of 22q11 is responsible for DiGeorge syndrome, velocardiofacial syndrome, congenital conotruncal heart defects, and related disorders. Familial transmission accounts for about 10-20 per cent of cases and most of the parents with deletions are nearly asymptomatic. This phenotypic variability makes it difficult to give appropriate genetic counselling. We report our experience on prenatal diagnosis of 22q11 deletion. We proposed prenatal detection of 22q11 microdeletion in 33 pregnancies. In two instances the parents refused prenatal diagnosis and one pregnancy ended spontaneously before the time of sampling. Fluorescent in situ hybridization (FISH) analysis on cultured amniotic cells with probes mapping in the commonly deleted region was used in the remaining 30 cases. Indications were classified into two groups. The first group included four couples with an abnormal family history of a deleted child and/or a deleted parent. No deletion was found in this group. The second one concerned pregnancies with a prenatally detected heart defect. Among these pregnancies with abnormal ultrasound findings, three deletions were found in the 26 samples tested. PMID- 9399352 TI - Maternal serum alpha-fetoprotein and human chorionic gonadotropin in pregnant women with acute parvovirus B19 infection with and without fetal complications. AB - Maternal serum alpha-fetoprotein (MS-AFP) and human gonadotropin (MS-hCG) were retrospectively determined in 137 sera from 65 pregnant women with an acute parvovirus B19 infection. In 35 of the pregnant women, fetal complications occurred (group 1); in the remaining 30 women, there were no fetal complications (group 2). In group I, significant elevations of MS-AFP were detected in 13 of the 35 women (37 per cent) and of MS-hCG in 25 of the 35 women (71 per cent). In nine of those women, sera were obtained before occurrence of fetal complications and MS-AFP was elevated in one case and MS-hCG in four cases. In one woman, both MS-AFP and MS-hCG were elevated. In group 2, significant elevations of MS-AFP were found in two of the 30 women (6-7 per cent) and of MS-hCG in five of the 30 women (16.7 per cent). Neither MS-AFP nor MS-hCG appears to be a regular early marker for poor pregnancy outcome in parvovirus B19-infected pregnancies, although they were frequently elevated at the time of complications. PMID- 9399354 TI - Ultrasound imaging of fetal neck anomalies: implications for the risk of aneuploidy and structural anomalies. AB - This study summarizes 24,000 transvaginal ultrasound examinations which were performed in a predominantly low-risk population at 14-16 weeks' gestation. 1254 (5.2 per cent) fetuses had a nuchal fold or a non-septated cystic hygroma. Of these fetuses, 140 (11.1 per cent) had additional structural anomalies. Cardiovascular anomalies were the most commonly detected structural malformations. Forty-three (3.4 per cent) fetuses were aneuploid. Trisomy 21 was the most common chromosomal anomaly (n = 27). Aneuploidy was significantly more common in fetuses who had a nuchal finding and an associated structural anomaly. The prevalence of nuchal fold and non-septated cystic hygroma, as well as the incidence of their associated structural anomalies, was similar. Based on these data, it is concluded that a complete ultrasonic survey of the fetus and karyotyping are advocated in fetuses with a nuchal abnormality, irrespective of maternal age or triple serum screening results. PMID- 9399353 TI - Prenatal diagnosis of congenital toxoplasmosis in 261 pregnancies. AB - Two hundred and sixty-one pregnant women underwent prenatal screening by cordocentesis and/or amniocentesis between 1987 and 1994. The following tests were used: (i) detection of anti-Toxoplasma gondii IgM, IgA, and IgE antibodies by immunocapture and the comparative immunological profile method based on enzyme linked immunofiltration assay of fetal blood and (ii) direct detection of the parasite in cell culture and by mouse inoculation with fetal blood (FB) and/or amniotic fluid (AF). Of the 31 cases of congenital toxoplasmosis, 24 (77 per cent) were detected prenatally. Overall, the FB and AF inoculation methods were the most effective (50 per cent sensitivity with FB inoculation to mice and/or cell culture and 74 per cent with AF). However, antibody detection in FB was the only positive test in three cases. Of 18 surviving children diagnosed prenatally, only one developed chorioretinitis (9 months of age). Seven newborns (23 per cent) with negative prenatal tests were diagnosed by postnatal laboratory monitoring, but none of these children developed clinical toxoplasmosis. There may have been more false negatives, as only 48 per cent of unaffected children were followed up for at least 12 months. All the tests had a specificity of 100 per cent. Fetal blood sampling has considerable value but also carries some risks and is currently being abandoned in favour of amniocentesis alone with gene amplification and mouse inoculation. PMID- 9399355 TI - A magnetic colloid system for isolation of rare cells from blood for fish analysis. AB - To improve cell recovery of trophoblast and nucleated red blood cells from maternal blood for diagnosis of chromosomal abnormalities, we have investigated the use of a magnetic sorting system that utilizes a ferrofluid. The main advantage of this system is that the beads used are small enough to allow visualization of chromosome-specific sequences by fluorescence in situ hybridization (FISH). The ferrofluid was validated using MAb340, a trophoblast specific antibody, and anti-CD71, used to sort for nucleated red blood cells. Antigen-positive cells could be efficiently sorted from a 1000-fold excess of antigen-negative cells and easily stained by FISH. We are currently evaluating its use on maternal blood samples. PMID- 9399356 TI - Expect the worse or hope for the best? Prenatal diagnosis of geleophysic dysplasia. AB - Geleophysic dysplasia is a rare, autosomal recessive disorder which causes disproportionate short stature associated with severe physical handicaps, but is compatible with survival into adulthood. We present a case, a first-born child, where genetic counselling difficulties arose following ultrasound recognition of short-limbed dwarfism in association with polyhydramnios and an initial incorrect prenatal diagnosis of lethal chondrodysplasia. After birth of the surviving affected infant, the parents had great difficulty accepting that there had been a prenatal misdiagnosis and they were greatly disappointed by our inability to predict the postnatal survival of an infant to whom no hope of life had previously been given. The correct diagnosis was not made until the proband was nearly 1 year old, and the true prognosis then became clearer. This experience underlines the relative ease of prenatal recognition of skeletal growth abnormalities compared with the considerable difficulties experienced in reaching a precise diagnosis. Thus, following prenatal diagnosis of unspecified chondrodysplasia when parents seek definite information about the prognosis, the temptation to be either overpessimistic or overoptimistic should be avoided. PMID- 9399357 TI - Prenatal diagnosis of a fetal head and neck neoplasm: differential diagnosis, management, and histology. AB - A fetal head and neck malignancy was prenatally diagnosed. The parents allowed the fetus to die during labour, due to the poor prognosis. We discuss the corresponding pathology findings, differential diagnosis, and management of this rare entity. Prenatal diagnosis of fetal neoplasms theoretically improves outcome, although this was not true in our case. PMID- 9399358 TI - Glycosaminoglycan analysis in amniotic fluid and in cultured fibroblasts from normal and holoprosencephalic human embryonic organs. AB - Glycosaminoglycans are polysaccharides involved in epithelial-mesenchymal interaction and cell differentiation and provide a meshwork which is essential to maintain a proper intercellular milieu. The development of embryonic organs can be accompanied by alterations in the glycosaminoglycan pattern. In pregnancies with malformed fetuses, there are alterations in total glycosaminoglycans and their components (chondroitin 4-6 sulphate, dermatan sulphate, and hyaluronic acid) in amniotic fluid. We examined total glycosaminoglycans and the percentage variations of the single classes in both amniotic fluid and culture medium of fibroblasts from heart, lung, and skin obtained from five normal human fetuses and one with holoprosencephaly. In the amniotic fluid total glycosaminoglycans and their sulphate classes were increased, whereas hyaluronic acid was decreased, compared with controls. The extracellular glycosaminoglycans showed hyaluronic acid reduction in skin, while chondroitin 4-6 sulphate plus dermatan sulphate and heparan sulphate were higher in skin and heart. Our data demonstrate that variations in the glycosaminoglycan pattern are associated with alterations of the cellular environment, which can prevent normal organogenesis. PMID- 9399359 TI - Prenatal ultrasonographic and molecular diagnosis of Apert syndrome. AB - Apert syndrome is a rare craniosynostosis syndrome with significant bilateral syndactyly of the hands and feet. Usually it is detected by ultrasonography during the third trimester unless there is a family history. We present an interesting sporadic case with features consistent with Apert syndrome detected as early as the first trimester. A first-trimester ultrasound evaluation prior to chorionic villus sampling (CVS) for maternal age 41 was within normal limits except for the suggestion of a 'mitten-like' hand and proximally placed thumb. Mid-trimester ultrasound was not diagnostic; however, following the development of polyhydramnios in the third trimester, the evaluation of the digits and facial features were strongly suggestive of Apert syndrome. Amniocentesis was performed and a molecular diagnosis of Apert syndrome was made and confirmed on cord blood. PMID- 9399360 TI - Maternal serum alpha-fetoprotein and human chorionic gonadotrophin in a pregnancy complicated by fetal sacrococcygeal teratoma. PMID- 9399361 TI - A 46,XX,der(13;14)(q10;q10),+21 child born after a 45,XX,der(13;14)(q10;q10) chromosomal finding in CVS. PMID- 9399363 TI - Current awareness in prenatal diagnosis. PMID- 9399362 TI - Maternal serum alpha-fetoprotein levels in congenital nephrosis. PMID- 9399364 TI - Benzodiazepine self-administration in humans and laboratory animals--implications for problems of long-term use and abuse. AB - Drug reinforcement may represent the primary behavioral-pharmacological mechanism underlying two types of problematic use of benzodiazepines--recreational abuse by polydrug abusers and inappropriate chronic use by patients. High dose polydrug abuse for the purpose of getting high is readily recognized as a significant social problem. Inappropriate chronic benzodiazepine use is more subtle but relatively common: for anxiolytics, 36% of past-year users (3% of the adult population in the US) report using these drugs for 4 consecutive months or longer. The risks of such long-term use are much better documented than the benefits. This paper provides a current review of various problems that have been identified with the long-term use and the recreational abuse of benzodiazepines, including memory impairment, risk of accidents, falls and hip fractures in the elderly, a withdrawal syndrome, brain damage, overuse in the elderly, overuse by chronic pain patients, overuse by alcoholics and recreational abuse among alcoholics and polydrug abusers. A comprehensive review of the literature on benzodiazepine reinforcing effects in humans and laboratory animals is also provided. Drug self-administration studies in humans and laboratory animals provide models of both types of problematic benzodiazepine use. Recreational abuse of benzodiazepines has been modeled in human research with polydrug abusers and in laboratory animal studies, which show that the reinforcing effect of benzodiazepines is intermediate relative to other sedative compounds and is increased in subjects with histories of previous sedative drug self administration. The problem of inappropriate long-term use of benzodiazepines by people without histories of drug abuse has been partially modeled in human studies showing that benzodiazepines function as reinforcers in subjects with anxiety, insomnia, and histories of moderate alcohol consumption, and in preclinical studies showing stable, low-rate benzodiazepine self-injection with concurrent physical dependence under conditions of continuous availability. Both human and animal research suggests that the drug history and current behavioral context may be important in the establishment of benzodiazepines as reinforcers. Limited human and animal research provides little support for the common belief that physical dependence enhances benzodiazepine reinforcement. PMID- 9399365 TI - Effects of cocaine and quinpirole on perceptual and motor functions in baboons. AB - The effects of cocaine and quinpirole were studied in baboons to determine whether quinpirole, a relatively selective D2/D3 dopamine agonist, produced effects similar to those of cocaine on perceptual and motor processes. To measure perceptual and motor function, three baboons were trained to discriminate differences between a standard vowel and four other synthetic vowels: response accuracy as well as response latencies, or "reaction times", were measured following drug administrations. Cocaine reduced reaction times in two baboons, and did not affect reaction times in a third; on the other hand, quinpirole lengthened reaction times in a dose-dependent manner in all baboons. Cocaine and quinpirole also differed in the time course to produce the maximal reaction time effect following drug administration. Cocaine and quinpirole did not differ consistently in their perceptual effects, as indicated by similar changes in d', a signal-detection index of discriminability. These distinct profiles of effects for cocaine and quinpirole suggest differing neurochemical actions for these two drugs. PMID- 9399366 TI - Anxiolytic and anticonvulsant activity of a synthetic neuroactive steroid Co 3 0593. AB - Endogeneously occurring neuroactive steroids, metabolites of progesterone and deoxycorticosterone, have been shown previously to interact with the GABAA receptor with great specificity in vitro and to have anticonvulsant, anxiolytic and sedative activity in vivo. However, these endogenously occurring steroids are not useful as therapeutic agents due to their potential metabolism to hormonally active steroids and their poor oral bioavailability. In an attempt to develop therapeutic agents which would maintain the pharmacological profiles of endogeneous neuroactive steroids but with increased oral bioavailability and reduced metabolic liability, we explored simple substitutions at the 3 beta position of the endogenous neuroactive steroid, 3 alpha-hydroxy-5 alpha-pregnan 20-one (3 alpha, 5 alpha-P). This report describes part of the in vitro and in vivo pharmacological profile of a 3 beta-substituted analog, 3 beta-ethenyl-3 alpha-hydroxy-5 alpha-pregnan-20-one (Co 3-0593). The compound exhibited anticonvulsant activity against pentylenetrazol-induced seizures in mice and rats (ED50 = 5.6 and 11.5 mg/kg, i.p., respectively). Co 3-0593 showed robust anxiolytic effects, comparable to benzodiazepines in the Geller-Seifter test after both SC and oral administration. Furthermore, the anxiolytic activity was maintained after chronic administration suggesting an absence of tolerance. The compound did not affect the acquisition of a learned response at both anticonvulsant and anxiolytic doses. However, at higher doses the compound showed rotorod deficit which was further enhanced by ethanol. In summary, 3 beta-ethenyl substituted 3 alpha, 5 alpha-P appeared to maintain the pharmacological activities of the endogenous neuroactive steroid with apparent oral activity. PMID- 9399367 TI - Blockade of pre- and post-synaptic 5-HT1A receptors does not modify the effect of fluoxetine or 5-hydroxytryptophan on ethanol and food intake in rats. AB - Selective serotonin reuptake inhibitors (SSRIs) or serotonin precursors inhibit ethanol and food intake by increasing the synaptic availability of 5-HT in the central nervous system. However, these agents can also increase 5-HT levels at somatodendritic 5-HT1A autoreceptors, with negative effects on serotonergic transmission. (+)WAY100135 [N-ter-butyl 3-4-(2-methoxy-phenyl) piperazin-1-yl-2 phenylpropanamide dihydrochloride] is a selective antagonist both at pre- and post-synaptic 5-HT1A receptors. The present study investigated the effect on ethanol and food intake of (+)WAY100135, given alone or coadministered with the SSRI fluoxetine or the 5-HT precursor 5-hydroxytryptophan (5-HTP) in genetically selected alcohol-preferring rats. Blockade of presynaptic 5-HT1A receptors after injection of (+)WAY100135, 0.1 or 1 microgram/rat, into the dorsal raphe did not significantly modify ethanol, food or total fluid intake. The same doses of (+)WAY100135 did not modify the inhibition of ethanol and food intake induced by intraperitoneal (i.p.) injection of fluoxetine, 5 mg/kg. Subcutaneous (s.c.) administration of (+)WAY100135 (1 or 10 mg/kg) did not affect the 3-h, or the overnight intake of ethanol, food or total fluids. Given together with i.p. fluoxetine (5 mg/kg) or s.c. 5-HTP (100 mg/kg plus carbidopa. 12.5 mg/kg), the same s.c. doses of (+)WAY100135 did not modify their inhibitory effect on ethanol and food consumption. Present findings suggest that blockade either of pre- or of pre- and postsynaptic 5-HT1A receptors does not potentiate the inhibitory effect of fluoxetine or 5-HTP on ethanol and food intake. PMID- 9399368 TI - Differential effects of ondansetron and alpha-flupenthixol on responding for conditioned reward. AB - Previous experiments have suggested that 5-HT3 antagonists such as ondansetron may alter reward-related behaviour that is dependent in part upon raised mesolimbic dopamine activity. However, the evidence for this is far from conclusive. One major behavioural role of dopamine is in the control of behaviour elicited by conditioned rewarding stimuli. To date, the effects of 5-HT3 antagonists on this function of mesolimbic dopamine have not been examined. Two experimental procedures were employed to examine the effects of ondansetron (10 and 100 micrograms/kg) on the acquisition of responding for conditioned reward, and on the response potentiating effect of intra-accumbens d-amphetamine (10 micrograms). These effects were compared to those elicited by the dopamine antagonist alpha-flupenthixol (0.1 mg/kg). In the first procedure, rats were trained to associate food pellet delivery with a conditioned stimulus (CS). Rats subsequently allowed to respond on a lever delivering this CS, and on an inactive lever, showed a greater preference for the lever delivering the CS, indicating that this CS functioned as a conditioned reward (CR). Ondansetron administered during the conditioning phase did not alter subsequent responding for the CR, but alpha-flupenthixol induced a small but significant reduction in responding on the CR lever. These results suggest that blockade of dopamine receptors, but not 5 HT3 receptors interfere with the learning of stimulus reward relationships. In the second procedure, d-amphetamine injected into the nucleus accumbens markedly potentiated responding for CR. Ondansetron at 10 micrograms/kg induced a small attenuation of this effect, without altering responding in its own right. However, at a higher dose (100 micrograms/kg) ondansetron plus amphetamine treatment significantly enhanced responding on the inactive lever. At both doses, the net effect of ondansetron was to produce a subtle impairment in the allocation of responses such that the differential responding on the CR versus NCR lever was diminished. In contrast to these effects alpha-flupenthixol significantly attenuated d-amphetamine's selective enhancement of responding for conditioned reward, as well as impairing the ability of the conditioned reward to elicit and maintain behaviour. These results confirm the role of dopamine in responding for conditioned reward, and suggest a possible modulators role for 5 HT3 receptors in this process. However, the effects of ondansetron on the acquisition of, and responding for, conditioned reward are clearly different from those induced by blockade of dopamine receptors. PMID- 9399369 TI - A behavioural analysis of the delayed non-matching to position task: the effects of scopolamine, lesions of the fornix and of the prelimbic region on mediating behaviours by rats. AB - The delayed non-matching to position task (DNMP) is a widely used automated test of spatial memory, yet its validity has been challenged by suggestions that animals use motor mediating behaviours which facilitate correct responding. This possibility was systematically studied by analysing video recordings of rats displaying delay-dependent and delay-independent deficits following lesions or drug manipulations. Rats were first trained to perform the DNMP task and whilst untreated, a number of potential mediating behaviours were identified from the video recorded behaviour. Two independent raters recorded any apparent motor strategies and attempted to predict the response the animals made during the choice phase of the task by viewing only behaviour during the delay periods. Subsequently, the behaviour of the same animals was examined following scopolamine treatment and following lesions of the prelimbic cortex or of the fornix. The experiment confirmed previous reports of delay-dependent and delay independent deficits under the varying conditions (drug, lesions), but also revealed that rats use clearly identifiable mediating behaviours that appear to facilitate correct responding in the DNMP task. Consequently, apparent "memory" impairments in the DNMP task, may reflect a disruption of behavioural strategies used by the animal to assist in performing the task. PMID- 9399370 TI - Ethanol attenuation of morphine dependence: comparison to dizocilpine. AB - Recent studies indicate that morphine dependence, assessed as the severity of naloxone-precipitated opiate withdrawal in rats, is attenuated by dizocipline, a non-competitive, excitatory amino acid antagonist. Because ethanol is a putative excitatory amino acid antagonist, the present study compared the effects of co administration of ethanol to that of dizocilpine on morphine dependence. Rats were administered morphine (10 mg/kg) twice daily for 9 days. One group received ethanol (1 g/kg) co-administration, another received dizocilpine (0.05 mg/kg) co administration, and a third served as vehicle controls. On day 10, all rats received naloxone (4 mg/kg) injections and ratings of several classic signs of opiate withdrawal were made. Both ethanol- and dizocilpine-treated rats showed significantly less severe precipitated opiate withdrawal overall, with the ethanol group showing reduced ratings of some specific signs. These results demonstrate that ethanol, like dizocilpine, attenuates the development of morphine dependence. The results are consistent with the action of ethanol at glutamate receptors. PMID- 9399371 TI - The effects of food deprivation and incentive motivation on blood glucose levels and cognitive function. AB - The current study investigated the relationships between blood glucose levels, mild food deprivation, sympathetic arousal, and cognitive processing efficiency. Subjects (n = 82) were randomly assigned to four experimental conditions, comprising combined manipulations of food deprivation and incentive motivation. Baseline and mid-session measurements of blood glucose, blood pressure and pulse rate were taken. Subjects completed a number of measures of cognitive processing efficiency and self report measures of affective and somatic state. Although glucose levels were lowered following food deprivation, there was no significant detrimental effect of food deprivation on task performance. However, improved recognition memory processing times were associated with deprivation. Incentive motivation was associated with faster simple reaction times and higher diastolic blood pressure. There were no significant relationships between glucose levels and task performance, further supporting the hypothesis that the brain is relatively invulnerable to short food deprivation. PMID- 9399373 TI - The mechanism of action of alpha 2 adrenoceptor blockers as revealed by effects on open field locomotion and escape reactions in the shuttle-box. AB - The precise mechanism of action of alpha 2 adrenoceptor blockers in not known, although in principle they have two main effects: (i) they stimulate the norepinephrinergic system by inhibiting the negative feed back of norepinephrine release (presynaptic effect) and (ii) they inhibit the effects of norepinephrine on postsynaptic alpha 2 adrenoceptors. We postulate that if the presynaptic actions of the antagonists prevail, the enhanced norepinephrine release leads to an activation of postsynaptic alpha 1 or beta adrenoceptors. In this case the effects of alpha 2 adrenoceptor blockers can be reversed by antagonists acting on the latter two adrenoceptors, since postsynaptic alpha 2 adrenoceptors are also blocked. If the postsynaptic blockade of alpha 2 adrenoceptors is the main cause of effects, than the blockade of alpha 1 or beta adrenoceptors should not reverse the action of alpha 2 blockers. The alpha 2 blocker idazoxan (dose 0.5-5 mg/kg) increased locomotor activity in an open field, an effect that was abolished by both alpha 1 and beta receptor blockers (prazosin and propranolol, respectively). Escape responses in a shuttle box were strongly suppressed by idazoxan (0.5-2 mg/kg). However, this effect was not changed by concomitant alpha 1 or beta receptor blockade. These results suggest that the mechanism of action of alpha 2 adrenoceptor blockers depends on which effects are studied. Exploration seems to be affected by a presynaptic mechanism as neurons bearing postsynaptic alpha 1 or beta adrenoceptors are involved in the control of this behavior, while escape reactions appear to be affected by the postsynaptic blockade of alpha 2 adrenoceptors (i.e. neurons bearing postsynaptic alpha 2 adrenoceptors are involved in its control). Thus, there is no generalized mechanism of action for alpha 2 adrenoceptor blockers; their precise mode of action should be investigated in each particular case. PMID- 9399372 TI - Effects of scopolamine on delayed-matching-to-sample and paired associates tests of visual memory and learning in human subjects: comparison with diazepam and implications for dementia. AB - Two experiments examined dose-related effects of 200, 400 and 600 micrograms scopolamine (n = 24, s.c.) and 5 and 10 mg diazepam (n = 6, PO) on parallel tests of visual memory and learning taken from the CANTAB battery. Scopolamine significantly impaired accuracy of performance on a delayed matching to sample test of visual recognition memory in a dose- and delay-dependent manner, but had only marginal decremental effects on a test of visuospatial paired associates learning. Scopolamine significantly lengthened decision times in a visual search matching to sample task at the 400 and 600 micrograms doses, without significantly affecting accuracy. The drug also impaired performance on tests of spatial (on accuracy and response time measures) and pattern (on response time only) memory. Most of the deleterious effects on scopolamine were removed by covariance analyses with indices of subjective sedation, but the effects of delayed matching accuracy and latency remained. By contrast, diazepam significantly impaired paired associates learning but affected delayed matching to sample in a delay-independent manner. These results suggest that scopolamine can produce selective deficits in tests of short-term visual recognition memory which do not depend on overall impairments in arousal and which contrast with deficits in visual associative learning produced by diazepam. They have implications for the pharmacological modelling of dementia and memory disorders in man and for the neurochemical substrates of the short-term recognition memory and associative learning for visual stimuli. PMID- 9399374 TI - Psychotropic drug intake in residents newly admitted to nursing homes. AB - While several surveys have shown that psychotropic drugs are frequently used by nursing home residents, no studies have been performed to investigate whether the rates of drug use increase during the stay in nursing homes or whether residents have taken these drugs already before admission. Therefore, we investigated 262 residents admitted to rural and urban nursing homes in Austria for prevalence of psychotropic drug intake before admission, shortly after admission, and 6 months later. Two weeks after admission, 72.1% of the residents were being treated with psychotropics, while 6 months later 79.0% were receiving these drugs. The significantly higher rates of psychotropic drug use among the psychiatrically ill and in those suffering from sleeping problems suggest that these drugs were prescribed aptly, but residents without appropriate criteria for drug intake were often also treated with psychotropics. During 3 months before admission to nursing homes, 45.5% of the sample reported having taken psychotropics. In more than half of residents without drug intake before admission, psychotropic treatment was initiated within the first 2 weeks after admission, while during the first 6 months after admission the rate of drug use increased only slightly. This suggests that a large percentage of psychotropic intake is due to nursing home orders. PMID- 9399375 TI - Behavioural and biochemical adaptations to nicotine in rats: influence of MK801, an NMDA receptor antagonist. AB - Chronic exposure of rats to nicotine can result in sensitization to the stimulant effects of nicotine on locomotor activity. At a biochemical level, chronic exposure to nicotine increases the number of CNS nicotinic binding sites, and this has been suggested as the basis for sensitization to nicotine. The present experiment was conducted to examine the effects of MK801, an NMDA receptor antagonist, on sensitization to nicotine. In addition, the hypothesis that MK801 may block behavioural sensitization by preventing the up-regulation of nicotinic receptors was tested by measuring receptor numbers in the same individuals using quantitative autoradiography with [3H]-cytisine and [3H]-MK801. Male Sprague Dawley rats were chronically treated with nicotine (0.4 mg/kg s.c.) or saline daily for 7 days. Over the next 2 days, in a counterbalanced order, rats were challenged with nicotine (0.4 mg/kg s.c.) or saline and locomotor activity was monitored. In saline-pretreated rats, nicotine produced a small increase in activity. Nicotine-pretreated rats exhibited higher levels of activity following a nicotine challenge. This sensitized response was attenuated in rats administered MK801 (0.3 mg/kg i.p.) 30 min before each daily nicotine injection. Rats pretreated with MK801 alone showed activity scores no different from saline pretreated control groups. Biochemical studies revealed increased [3H]-cytisine binding following chronic nicotine treatment; however, receptor increases were significantly attenuated by MK801 pretreatment. Binding of [3H]-MK801 remained unchanged across the four groups. The results suggest that MK801 prevents behavioural sensitization to nicotine via the prevention of receptor up regulation. Although the findings support the notion that receptor up-regulation may be the basis for the increased responsiveness to nicotine, other interpretations are possible. PMID- 9399376 TI - Genetics, haloperidol-induced catalepsy and haloperidol-induced changes in acoustic startle and prepulse inhibition. AB - The acoustic startle response (ASR), prepulse inhibition (PPI) of the ASR and the effects of haloperidol on the ASR and PPI were examined in C57BL/6J (B6) and DBA/2 (D2) inbred mouse strains and their F1 and F2 progeny. The startle stimulus was a 60-ms, 110-dB, 10-kHz tone; the prepulse stimuli were 20-ms, white noise bursts at 56, 68 and 80 dB against a 50-dB background presented 100-ms before the startle pulse. The B6 strain showed modest PPI (25-40%); in contrast, the D2 strain showed on average no PPI and numerous individuals showed prepulse augmentation (PPA). The F2 progeny showed an intermediate PPI; however, the extreme values ranged from 200% PPA to essentially 100% PPI. Haloperidol in dose dependent fashion, increased PPI in both the B6 and D2 strains; the threshold dose was in the range of 0.1-0.2 mg/kg. Raclopride (0.3 mg/kg), clozapine (2 mg/kg) and risperidone (0.4 mg/kg) also increased PPI in both strains. The effects of haloperidol (0.4 mg/kg) on PPI in 140 F2 progeny were examined. For all prepulse intensities, there were highly significant (r > 0.80) and negative correlations between baseline PPI and the haloperidol-induced change in PPI. Thus, those animals that showed the greatest PPA showed the greatest haloperidol induced increase in PPI. There was, however, significant variance in the haloperidol response; plots of the regression residuals showed the most and least responsive animals differed by almost 100% in effect on PPI. The F2 progeny were subsequently phenotyped for haloperidol-induced catalepsy. There was no association between the variation in effects on catalepsy and PPI. However, it was observed that those individuals with the poorest baseline PPI were catalepsy non-responsive. PMID- 9399377 TI - Sexual segregation in infant mice: behavioural and neuroendocrine responses to d amphetamine administration. AB - Individual differences arise from both genetic and epigenetic factors. The aim of this study was to test whether pups raised in distinct socio-sexual conditions would show different behavioural and neuroendocrine responses to d-amphetamine (AMPH) administration upon placement in a novel environment. This issue was addressed by testing infant CD-1 mouse pups of both sexes at three different developmental ages [3, 8, or 18 postnatal (PND) days]. These pups were raised from birth in all-male, all-female, or mixed-sex litters. AMPH effects were assessed as a function of the hypothalamic-pituitary-adrenal (HPA) axis activational state using litters that were either maternally deprived for 24 h (DEP) or normally kept with the dam (NDEP). A concomitant maternal behaviour score carried out on selected postpartum days showed that mothers taking care of all-male litters were more often involved in Active nursing than those rearing the mixed-sex ones, whereas the latter were found more often Laying still out of the nest. Basal and stress-induced corticosterone (CORT) secretion was increased in unisexually reared pups following maternal deprivation, an effect limited to PND 3. In general, neuroendocrine and behavioural responses to AMPH were found to be dissociated and were affected by sexual segregation only in conjunction with maternal deprivation. On PND 3, AMPH injection (1 or 3 mg/kg, i.p.) decreased CORT secretion in deprived unisexually reared subjects without affecting their behaviour. As a whole, behavioural changes due to unisexual rearing were limited to female subjects. On PND 8, unisexually reared females showed, upon maternal deprivation, a generalized shift to the left in the dose-response curve to AMPH for Crossing behaviour, while on PND 18 AMPH-induced stereotypies were considerably reduced in sexually segregated females, especially following maternal deprivation. Thus, maternal deprivation appeared to "sensitize" the monoaminergic system to an AMPH challenge. The individual behavioural and neuroendocrine profiles shown in response to a stressful challenge suggest that changes in social stimulation early during development might produce subtle shifts in the function of selected central monoaminergic systems. PMID- 9399378 TI - Treatment of neuroleptic-induced akathisia with nicotine patches. AB - We administered 14 mg nicotine patches to 16 patients, all non-smokers, who displayed akathisia from antipsychotic drugs. On single-blind ratings, akathisia appeared significantly reduced on days when patients were wearing the patches as compared to the baseline day. These findings, if confirmed, may help to explain the high rates of tobacco use among psychotic patients, and may suggest avenues for the treatment of akathisia. PMID- 9399379 TI - A time-activity baseline to measure pharmacological and non-pharmacological manipulations of PCP-induced activity in mice. AB - At critical doses of PCP (10 mg/kg i.p. in the present studies), locomotor stimulation in mice is initially suppressed by short-lasting ataxia, albeit at higher levels of activity than controls. This provides a time-activity baseline of PCP-stimulated locomotion potentially sensitive to i) pharmacological antagonism indicated by a change in the time-activity relationship to that seen at lower PCP doses, ii) interaction with the ataxic phase resulting in further decreases in activity similar to that seen at higher PCP doses and iii) reductions in activity without a change in the time-activity relationship. This baseline was explored using three manipulations employed in the clinical management of PCP toxicity: treatment with a neuroleptic (haloperidol), a benzodiazepine (chlordiazepoxide) and modification in environmental stimulation (changing of lighting conditions). Both haloperidol (0.125-0.5 mg/ kg, i.p.) and chlordiazepoxide (5-20 mg/kg i.p.) further reduced activity during the ataxic phase of the PCP time-activity relationship qualitatively similar to the effects of pentobarbital (20-40 mg/kg i.p.). Changing of lighting conditions from red to white light resulted in significant reductions in levels of activity of PCP treated animals but no change in the time-activity relationship. No manipulation resulted in true reversal of the PCP induced time-activity relationship. The results parallel the clinical findings that neuroleptic and benzodiazepine administration have no specific effects upon PCP-intoxication and that environmental manipulation may modify the degree of PCP stimulation. The time activity baseline described may prove useful in the evaluation of the effects of pharmacological and non-pharmacological manipulations of PCP-induced activity in rodents. PMID- 9399380 TI - Effects of hot tea, coffee and water ingestion on physiological responses and mood: the role of caffeine, water and beverage type. AB - Psychopharmacological studies using caffeinated beverages or caffeine have rarely considered temporal effects on psychological and physiological function or the specific contribution of caffeine, hot water, or beverage type to the observed effects. The effect of 400 ml hot tea, coffee, and water consumption on systolic and diastolic blood pressure (SBP and DBP), heart rate, skin conductance (a measure of sympathetic nervous system activation), skin temperature, salivary cortisol, and mood were monitored in 16 healthy caffeine-withdrawn (14 h) subjects in a complete crossover design. Beverages were ingested with/without 100 mg caffeine and milk (tea/coffee only). Hot beverage ingestion rapidly increased skin conductance and temperature (+1.7 degrees C) with peak effects observed only 10-30 min post-consumption. Caffeine in the beverage rapidly augmented skin conductance responses but, in contrast to the effect of hot water, reduced the skin temperature response and increased SBP (+2.8 mmHg) and DBP (+2.1 mmHg) 30-60 min post-consumption. Both caffeine and milk addition to beverages independently improved mood and reduced anxiety 30 and 60 min post-consumption. Milk addition had no other effects apart from attenuating the transient increase in physiological responses associated with the drinking phase. There were no effects of beverage consumption on salivary cortisol or of beverage vehicle on salivary caffeine levels, the latter indicating that caffeine pharmacokinetics was similar in both tea and coffee, and not different from caffeinated water. In keeping with this, the responses to tea and coffee ingestion were similar and largely accounted for by the effects of hot water and caffeine. However, tea potentiated the increase in skin temperature compared to coffee and water indicative of a greater vasodilatory response plausibly related to the presence of flavonoids in tea. We conclude that ingestion of hot caffeinated beverages stimulates physiological processes faster than hitherto described, primarily via the effects of hot water and caffeine, but with beverage type and milk playing important modulatory roles. PMID- 9399381 TI - Corticotropin-releasing factor binding sites in cortex of depressed suicides. AB - Corticotropin-releasing factor (CRF) receptors were measured by saturation binding in frontal and motor cortex of suicides with a firm retrospective diagnosis of depression, and matched controls. The suicides were divided into those who were free of antidepressant drugs, and those in whom prescription of antidepressant drugs was clearly documented. There were no differences in the number or affinity of CRF receptors between antidepressant-free or antidepressant treated suicides and matched controls in either brain region. When suicides were divided according to violence of death, again there were no differences between violent or non-violent suicides and controls. PMID- 9399382 TI - Effect of central 5-hydroxytryptamine depletion on performance in the "time-left" procedure: further evidence for a role of the 5-hydroxytryptaminergic pathways in behavioural "switching". AB - This experiment examined the effect of destruction of the ascending 5 hydroxytryptaminergic (5HTergic) pathways on performance in a free-operant timing schedule: the "time-left" procedure. Rats received either injections of 5,7 dihydroxytryptamine into the dorsal and median raphe nuclei or sham lesions. They were trained in a discrete trials schedule in which reinforcers were provided for responding on either of two levers, A and B. At a random time point, t s after the start of each trial, a response on A resulted in the delivery of one food pellet after dA s, whereas a response on B resulted in the delivery of two pellets after 60-t s. The value of dA was varied between 1 and 8 s in different phases of the experiment. Both groups showed decreasing response rates on lever A and increasing response rates on lever B as a function of time within the trial. An index of timing (T75: the time within the trial at which relative response rate on B attained a value of 75%) was systematically related to the value of dA, but did not differ significantly between lesioned and control rats. However, the lesioned group showed significantly higher rates of switching between response alternatives than the sham-lesioned group at all values of dA. The levels of 5HT and 5-hydroxyindoleacetic acid were reduced in the brains of the lesioned rats, but the levels of noradrenaline and dopamine were not significantly altered. The results provide further evidence that the ascending 5HTergic pathways may contribute to the inhibitory regulation of switching between behavioural states. PMID- 9399383 TI - Nicotine-induced decreases in VTA electrical self-stimulation thresholds: blockade by haloperidol and mecamylamine but not scopolamine or ondansetron. AB - The effects of repeated daily injections of (-)-nicotine (+) hydrogen tartrate (mg kg-1 s.c.) on electrical self-stimulation of the ventral tegmental area were investigated. Nicotine reduced the frequency required to maintain half-maximal response rates with animals responding in rate-frequency threshold tests. Under these conditions, nicotine induced an increase in the total number of self stimulation responses per session, but had no statistically significant effects on the maximal response rate. These effects of nicotine were observed by the second day of administration of this drug. Acute injections of the D2-like dopamine receptor antagonist haloperidol (0.03 mg kg-1 s.c.) and of the nicotinic acetylcholine receptor antagonist mecamylamine (1 mg kg-1 s.c.) attenuated the effects of nicotine, indicating that the observed effects involve stimulation of D2-like dopamine receptors as a result of nicotinic receptor activation. The muscarinic acetylcholine receptor antagonist scopolamine (3 mg kg-1 s.c.) and the serotonin 5-HT3 receptor antagonist ondansetron (0.01 and 0.1 mg kg-1 s.c.) did not alter the effects of nicotine. The results of this study indicate that repeated daily administration of (-)-nicotine increases the rewarding effects of electrical self-stimulation of the ventral tegmental area. These data are consistent with the proposal that repeated daily injections of nicotine positively effect a mesolimbic dopaminergic substrate of reward. PMID- 9399384 TI - Locomotor sensitization to quinpirole: environment-modulated increase in efficacy and context-dependent increase in potency. AB - This study examines whether behavioural sensitization to the dopamine agonist, quinpirole, reflects an increase in the drug's potency and/or efficacy to induce locomotion, and how these parameters are influenced by environmental context. Three experiments were conducted in which animals received either chronic quinpirole (10 x 0.5 mg/kg, twice weekly) or saline injections in either the home cage environment, an alternate environment or the testing environment (activity monitors), followed by a dose-response test for the expression of sensitization in the activity monitors. Compared to the acute dose-response relationship, chronic quinpirole increased the maximum response. This increase in efficacy was significantly higher in animals treated with quinpirole in a non-home cage environment compared to those that received chronic treatment in the home cage. A leftward shift in the dose-effect function was observed only in animals with prior drug experience in the testing environment. Results indicate that locomotor sensitization to quinpirole reflects an environment-modulated increase in the drug's efficacy, and an environment-dependent increase in drug potency. Efficacy and potency may be subject to sensitization by non-associational and associational mechanisms, respectively. PMID- 9399385 TI - Dopaminergic mediation of the discriminative stimulus effects of bupropion in rats. AB - Bupropion is a novel, non-tricyclic antidepressant with a primary pharmacological action of monoamine uptake inhibition. The drug resembles a psychostimulant in terms of its neurochemical and behavioural profiles in vivo, but it does not reliably produce stimulant-like effects in humans at clinically prescribed doses. Bupropion binds with modest selectivity to the dopamine transporter, but its behavioural effects have often been attributed to its inhibition of norepinephrine uptake. This experiment examines monoaminergic involvement in the discriminative stimulus effects of bupropion. Rats were trained to press one lever when injected i.p. with bupropion (17.0 mg/kg), and another lever when injected with saline. In substitution tests, dose-response curves were obtained for several monoamine uptake inhibitors. Nine of ten dopamine uptake blockers fully substituted for bupropion; the exception, indatraline (LU 19-005), partially substituted (71% bupropion-appropriate responding). Serotonin and norepinephrine uptake blockers (zimelidine and nisoxetine, respectively) produced negligible or limited substitution, and the anti-muscarinic dopamine uptake blocker benztropine produced limited partial substitution. A series of dopamine D1-like and D2-like receptor agonists were also tested: only the D2-like agonist RU 24213 fully substituted; three other D2-like agonists and four D1-like agonists partially substituted (50% < drug responding < 80%). Antagonism of the discriminative effects of bupropion was obtained with a D1- and a D2-like dopamine antagonist. The results demonstrate strong similarities with those obtained using other dopamine uptake inhibitors as training drugs, and support the view that the behavioural effects of bupropion are primarily mediated by dopaminergic mechanisms. PMID- 9399386 TI - The significance and potential molecular mechanisms of gastrointestinal barrier homeostasis. PMID- 9399387 TI - Quality of life in patients with different types of functional constipation. AB - BACKGROUND: Little information is available about the health-related quality of life (QoL) in patients with different types of chronic constipation. METHODS: We used two self-administered questionnaires, the Psychological General Well-Being (PGWB) index and the Gastrointestinal Symptom Rating Scale (GSRS) to assess QoL and gastrointestinal symptoms in 102 consecutive patients with chronic constipation. The type of constipation was determined from transit time, electrophysiologic investigation of sphincter function, anorectal manometry, and defecography. RESULTS: Overall, our patients with constipation reported low scores for general well-being (mean score, 85.5, compared with 102.9 in a healthy population). Patients with normal-transit constipation (n = 49) reported considerably lower scores in the PGWB than those with slow-transit constipation (n = 35). The symptoms increased frequency of defecation, loose stools, and urgent need for defecation were commoner in normal-transit constipation, which indicates that this group may have a relation to the irritable bowel syndrome. The overall PGWB index was strongly correlated with the total GSRS (P < 0.001). CONCLUSIONS: The general well-being of patients with chronic constipation is lower than that of a comparable normal population. Symptom severity correlates negatively with perceived quality of life. PMID- 9399388 TI - Osteoporosis after total gastrectomy. Results of a prospective, clinical study. AB - BACKGROUND: Osteopenia and enhanced risk of fractures have been reported after partial gastrectomy, but the significance of total gastrectomy is still unknown. METHODS: Twenty-six patients were followed up for at least 3 years after total gastrectomy. The intake and S-levels of vitamin D, phosphate, magnesium, and calcium were prospectively studied, and a whole-body dual-energy X-ray absorptiometry scan was performed at a mean of 5 years after gastrectomy. RESULTS: At this time point we found normal blood levels of vitamin D, calcium, and phosphate. Food intakes of phosphate, calcium, magnesium, and vitamin D reached the recommended daily allowances. Bone mineral density was similar to that of a control population, and increasing values were seen concomitant with an increase in body weight with the time after gastrectomy. CONCLUSIONS: Calcium homeostasis and bone mineral densities seem not to be affected by total gastrectomy, at least when studied over a period of 5 years, an observation that hypothetically can be explained by weight recovery with time after the operation. PMID- 9399389 TI - Use of commonly prescribed antibiotics is not associated with prevalence of Helicobacter pylori infection in adults. AB - BACKGROUND: The aim of the study was to investigate the association of the use of commonly prescribed antibiotics with prevalence of Helicobacter pylori infection in a population of adult outpatients. METHODS: All patients aged 15-79 years who visited the practice of a general practitioner (GP) between June and September 1996 in a suburban community near Ulm, a city in southern Germany, were asked to participate in the study. Infection status was determined with a 13C-urea breath test. In addition, the patients were asked to fill out a self-administered questionnaire. RESULTS: Overall, 475 outpatients were included in the study (response, 94.1%). A total of 266 patients (56.0%) reported a history of antibiotic treatment within the past 5 years, whereas 147 patients (30.9%) did not (62 patients (13.1%) did not know). Prevalence of infection in patients with a history of antibiotic medication during the past 5 years was 23.3%, whereas the prevalence of infection was 20.4% in subjects without antibiotic treatment (P = 0.283 after stratification for age). Control for other potential confounders by multivariable analysis did not materially alter the results. CONCLUSION: Coincidental antibiotic treatment is not associated with H. pylori prevalence in adults. PMID- 9399390 TI - The effect of cisapride in maintaining symptomatic remission in patients with gastro-oesophageal reflux disease. AB - BACKGROUND: Successful treatment of gastro-oesophageal reflux disease (GORD) has traditionally been assessed as healing of reflux oesophagitis, which may not be relevant in patients with moderate disease. In these patients symptom relief and patient satisfaction with therapy are of fundamental importance. Cisapride has well-documented prokinetic effects and may be well suited for long-term therapy of GORD, but its effectiveness in purely symptomatic treatment is unknown. We therefore compared two dosage regimens of cisapride with placebo over a period of 6 months in patients with evidence of gastrooesophageal reflux, initially treated with antisecretory medication, with regard to maintaining symptom relief and satisfaction with treatment. METHODS: Five hundred and thirty-five patients with reflux oesophagitis grade 1 (n = 293) or 2 (n = 124) or with no reflux oesophagitis but pathologic 24-h pH-metry (n = 118) achieved satisfactory symptom relief with an H2-receptor antagonist or proton pump inhibitor within 4-8 weeks. In a double-blind randomized, parallel-group study, they were then treated with cisapride, 20 mg at night or 20 mg twice daily, or placebo and followed up for a maximum period of 6 months. Relapse was defined as dissatisfaction with therapy or an average consumption of more than two antacid tablets a day. RESULTS: Median time to relapse was 63 days for cisapride, 20 mg twice daily; 59 days for cisapride, 20 mg at night; and 49 days for placebo. Time to relapse was not significantly different (P = 0.09). Presence and grade of oesophagitis at base line, type of therapy before randomization, and pattern of non-reflux symptoms at base line did not influence these findings significantly. CONCLUSION: The study indicates that cisapride is of limited value in maintenance therapy of GORD in patients in whom symptom relief has been accomplished with potent antisecretory medication. This 'step-down' approach to therapy seems disadvantageous in the long-term therapy of GORD. PMID- 9399391 TI - Gastrointestinal motility disorders in patients with inactive Crohn's disease. AB - BACKGROUND: Although some symptoms of Crohn's disease may be related to gastrointestinal motility disorders, studies on gastrointestinal motility in inactive Crohn's disease are lacking. METHODS: Fasting and postprandial motor activity (1 h) was recorded in the gastric antrum and upper small intestine of 35 patients with inactive Crohn's disease and 18 controls, using conventional manometry. RESULTS: Motor disorders were observed in 26 of 35 patients. The number of phase-II contractions was reduced (1.3 +/- 0.7/min versus 1.8 +/- 0.6/min in controls; P < 0.02) (mean +/- standard deviation), whereas the incidence of propagated single (2.2 +/- 3.2/h versus 0.5 +/- 0.6/h; P < 0.03) and clustered contractions (3.8 +/- 7/h versus 1.1 +/- 1.4, P < 0.04) was markedly increased. Motor abnormalities were more frequent and severe in patients with Crohn's ileitis than in controls, and in patients with gastrointestinal symptoms than in asymptomatic patients. CONCLUSION: Most patients with inactive, uncomplicated Crohn's disease show marked gastrointestinal motor disorders, characterized either by reduced incidence of small-bowel contractions and increased incidence of single or clustered propagated contractions. PMID- 9399392 TI - Effects of insulin-like growth factor-I administration on radiation enteritis in rats. AB - BACKGROUND: Acute radiation-induced damage to the small bowel occurs frequently during abdominal radiotherapy. Since the small intestine is selectively responsive to the growth-promoting effects of insulin-like growth factor-I (IGF I), we investigated the effects of IGF-I administration on mucosal recovery from radiation enteritis in the rat. METHODS: Rats received a single 10-Gy dose of total abdominal irradiation followed by implantation of mini-pumps infusing either IGF-I or vehicle for 4 days. After the rats had been killed, gut organs were weighed before light microscopic and biochemical examination. RESULTS: Irradiated rats receiving IGF-I lost less body weight than vehicle-treated rats, whereas the wet weights of the stomach, small intestine, and colon were increased by 10%, 19%, and 21%, respectively, and crypt depth was increased in the duodenum, jejunum, and ileum. CONCLUSIONS: IGF-I administration after abdominal irradiation increased small-intestinal mass and improved indicators of mucosal integrity, suggesting acceleration of small-intestinal mucosal recovery from radiation injury. PMID- 9399393 TI - Intestinal obstruction after appendectomy. AB - BACKGROUND: The frequency of intestinal obstruction varies in the literature (0.2 10.7%) and requires evaluation in a proper design. METHODS: From 1978 to 1985, 1951 patients underwent appendectomy; 58 patients were excluded because of appendectomy per occasionem, 156 because of previous laparotomy, and 190 because of simultaneous major surgery. Three foreigners were lost to follow-up. The cohort was linked to the Danish National Inpatient Register for identification of cases, defined by intestinal obstruction requiring surgical intervention. RESULTS: The follow-up period was long (median, 3563 days; range, 2-5113). Twenty one patients developed intestinal obstruction. The cumulated incidence was 0.33% after 30 days, 0.79% after 1 year, and 1.51% after 14 years. Female sex as compared with male sex (RR = 3.91; 95% confidence limits (CL), 1.25-12.0) and removal of a removal of a normal appendix as compared with an inflamed appendix (RR = 4.0; 95% CL, 1.28-12.5) carried a significantly higher risk of intestinal obstruction. CONCLUSION: Intestinal obstruction after open appendectomy is rare. PMID- 9399394 TI - High incidence and prevalence of adult coeliac disease. Augmented diagnostic approach. AB - BACKGROUND: The diagnosis of coeliac disease is easily overlooked as patients can present with mild or atypical symptoms, or the condition can even be clinically silent. Our aim was to detect coeliac disease patients with such atypical or no symptoms as well as those with typical features. METHODS: The incidence of adult coeliac disease in Tampere was calculated from 1975 to 1994 and the prevalence as of 31 December 1994. Open-access endoscopy was available for general practitioners, and small-bowel biopsy was done routinely. Serologic screening was applied to patients with an increased risk of coeliac disease. RESULTS: The incidence of coeliac disease increased tenfold, and the prevalence was 270 per 100,000 inhabitants in 1994. Twenty per cent were found by serologic screening and 10% as a result of routine biopsy; 24% had dermatitis herpetiformis. CONCLUSIONS: Our diagnostic approach gave a coeliac prevalence similar to that found in population screening studies. One-third had silent coeliac disease. PMID- 9399395 TI - Inflammatory bowel diseases: health care and costs in Sweden in 1994. AB - BACKGROUND: Inflammatory bowel disease (IBD) has highly variable course and severity. Since comprehensive data on its impact are scarce, we analyzed all IBD care and costs in Sweden (population, 8.8 million). METHODS: A cross-sectional observational study, using national registers and surveys on ambulatory care, hospital admissions, medication, sickness leave, and early retirement for IBD in 1994, was carried out. We calculated direct health care costs and indirect 1-year costs caused by morbidity. RESULTS: Ambulatory care was concentrated to specialists in internal medicine at hospitals. One-fourth of the patients accounted for 48% of 1994 hospital admissions. Medication was predominantly aminosalicylates and steroids. Sickness leave episodes were long--on average, 6 weeks. Although uncommon, early retirements lasted 14 years on average. With regard to the underlying prevalence, the use of health care and compensations by Crohn's disease patients was two to four times that of patients with ulcerative colitis. Morbidity took 68% of total costs. Among direct costs, admissions accounted for 58%. Neither complications nor surveillance added much. CONCLUSION: Ulcerative colitis is twice as common as Crohn's disease. In health care use, these roles are reversed. Since morbidity causes two-thirds of the costs, comprehensive analyses, including indirect costs, are necessary when evaluating new diagnostics and therapies. PMID- 9399396 TI - Prevalence of Helicobacter pylori infection and related upper gastrointestinal lesions in patients with inflammatory bowel diseases. A cross-sectional study with matching. AB - BACKGROUND: Although a reduced prevalence of Helicobacter pylori infection has been observed in inflammatory bowel disease (IBD) patients, the clinical significance of H. pylori infection in this setting remains unknown. The aim of this study was, therefore, to evaluate the prevalence of H. pylori infection in a large series of IBD patients and the frequency of gastroduodenal lesions in those who agreed to undergo upper GI endoscopy. METHODS: Two hundred and sixteen consecutive IBD patients (123 with Crohn's disease (CD) and 93 with ulcerative colitis (UC)) had their anti-H. pylori IgG titres measured. Two hundred and sixteen blood donors matched for age, sex, place of birth in Italy, and socioeconomic status served as controls. All patients were offered the possibility of undergoing endoscopy with antral and corpus biopsies regardless of their H. pylori status. RESULTS: The overall seroprevalence of H. pylori infection was 48% in IBD patients versus 59% in the control group (P < 0.05), with a significantly lower frequency in CD versus UC patients (41% versus 56%). After adjustment for age, education, and socioeconomic status CD remained associated with a significantly lower risk of H. pylori infection. Previous therapy with sulphasalazine but not with 5-aminosalicylic acid or with steroids/immunosuppressants was associated with a reduced risk of H. pylori infection both in CD and UC patients. One hundred and eighty-nine patients (110 with CD and 79 with UC) underwent endoscopy; the prevalence of peptic ulcer was similar in both groups (5.5% in CD and 5.1% in UC patients); however, 11 more CD patients had gastroduodenal ulcers that were interpreted as CD-related; 7 of these patients had never had foregut symptoms. Two CD patients had granulomatous gastritis at histology, and another 16 patients with CD had H. pylori-negative gastritis. CONCLUSIONS: IBD patients have a reduced prevalence of H. pylori infection as compared with matched healthy controls; this appears mostly attributable to a reduced frequency of H. pylori colonization in CD patients. Previous use of sulphasalazine is associated with a reduced risk of infection both in CD and UC patients. Of CD patients 10% have a gastroduodenal localization of their disease, which is often asymptomatic. Of CD patients 15% also have H. pylori-negative gastritis at histology. PMID- 9399397 TI - Detection of p53 autoantibodies in sera of gastric cancer patients and their prognostic relevance. AB - BACKGROUND: Changes in the p53 gene product can be immunogenic and enable the formation of p53 serum antibodies (p53ab), detectable in patients with different cancer types. So far, there have been no reports describing the detectability of p53ab in gastric cancer patients. METHODS: We investigated the presence of p53ab and their clinical relevance in a cohort of 74 gastric cancer patients, using an enzyme-linked immunosorbent assay system. RESULTS: In our investigation 20.3% of all patients (15 of 74) and 46.9% of the patients with immunohistochemically (IHC) p53-positive tumors (15 of 32) showed detectable p53ab in serum. All p53ab positive patients had IHC p53-positive tumors. We have found a significant correlation of p53ab with a higher tumor stage (P = 0.002) and also with a poor prognosis of survival (P = 0.04). CONCLUSION: We have shown that in gastric cancer patients p53ab are also detectable and that p53ab positivity is a predictor of an unfavorable prognosis. PMID- 9399398 TI - The anti-proliferative effect of plasma from rats with acute fulminant hepatic failure. AB - BACKGROUND: During fulminant hepatic failure (FHF) metabolites normally cleared by the liver accumulate in the circulation and cause hepatic coma. It is believed that the plasma of FHF patients has an inhibitory effect on liver regeneration. Plasma exchange was used to study the effect of plasma collected from donor FHF rats on liver regeneration in two-thirds partially hepatectomized syngeneic animals. METHODS: FHF and hepatic coma were induced in donors by administration of galactosamine at a dose of 1.85 g/kg. Plasma from donors in either grade-II or -IV coma was transfused by plasma exchange into partially hepatectomized animals 2h after resection. RESULTS: The livers from donor animals showed evidence of oval cell activation 1-2 days after galactosamine, but differentiation of oval cells to hepatocytes did not occur before the development of coma. The plasma collected from animals in grade-IV coma totally abolished regeneration in the partially hepatectomized recipients. CONCLUSION: These results support the hypothesis that metabolites present in the plasma during FHF inhibit liver regeneration. PMID- 9399399 TI - Frequent occurrence of non-specific gliadin antibodies in chronic liver disease. Endomysial but not gliadin antibodies predict coeliac disease in patients with chronic liver disease. AB - BACKGROUND: The frequency of gliadin antibody (GA) positivity has been found to be increased among patients with chronic liver disease, as has that of coeliac disease (CD). CD has also been found to be increased among patients with primary biliary cirrhosis (PBC) or primary sclerosing cholangitis (PSC). METHODS: To investigate these relationships further, a micro-enzyme-linked immunosorbent assay and immunofluorescence tests for GAs and endomysial antibodies (EMAs) were performed in large subgroups of patients representing various chronic liver diseases and in healthy blood donors. RESULTS: As compared with blood donors (among whom it was 5%) the frequency of IgA GA positivity was higher in all patient subgroups: alcoholic liver disease, 20% (22 of 110, P < 0.001); PBC, 16% (16 of 101, P < 0.001); PSC, 24% (19 of 80, P < 0.001); chronic hepatitis, 19% (13 of 70, P < 0.001); and hepatitis C virus infection, 11% (11 of 104, P < 0.01). Two patients with autoimmune chronic hepatitis were EMA-positive, and in both cases the presence of CD was verified by small-bowel biopsy. CONCLUSIONS: IgA GA positivity generally occurs at increased frequency among patients with chronic liver disease and may represent non-specific immune activation. In liver disease GA testing is not useful in screening for CD, whereas the EMA test seems to be highly specific. CD is more prevalent than expected among patients with autoimmune chronic hepatitis but not among those with PBC or PSC. PMID- 9399400 TI - Percutaneous ethanol injection in the treatment of hepatocellular carcinoma. A multicenter survey of evaluation practices and complication rates. AB - BACKGROUND: Percutaneous ethanol injection (PEI) has become a widely used procedure in the treatment of hepatocellular carcinoma (HCC). However, the criteria for selecting patients are not standardized, and little information is available about the complications of the procedure. METHODS: A questionnaire was sent to 11 experienced Italian centers. It investigated: the size and the number of HCC nodules suitable for treatment and the Child-Pugh risk class of the associated cirrhosis; the performance of the procedure; the number and characteristics of the patients treated; and, finally, any complications. RESULTS: Most of the centers performed PEI in single HCC nodules less than 5 cm in diameter or in multiple nodules if fewer than three, the larger being less than 3 cm. Patients in Child-Pugh's classes A, B, and C with single nodules were generally considered for PEI. A prothrombin time of less than 40% and a platelet count of less than 40,000/mm3 contraindicated PEI in most of the centers. PEI was generally performed on outpatients, using Chiba or spinal needles. One thousand and sixty-six patients (8118 sessions) were enrolled; 74% had a single HCC nodule and 26% multiple nodules. All except four had cirrhosis; 53% were in Child class A, 38% in class B, and 9% in class C. The mean number of sessions needed to destroy an HCC nodule was 6.7 (range, 2-14), with a mean alcohol injection volume of 5.0 ml per session (range, 2-20 ml). One death (0.09%) and 34 complications (3.2%) were reported. Among the complications we call attention to the hemorrhagic ones (eight cases) and tumoral seeding (seven cases). Severe pain experienced during the maneuver led to discontinuation of the procedure in 3.7% of the patients; 13.5% of the patients required analgesics and 24% had fever after PEI. CONCLUSIONS: Some procedural aspects of PEI treatment differ among the various centers a standardization is advisable. In the present survey PEI is a low-risk technique. PMID- 9399402 TI - Salmonellosis: an unusual complication of hepatocellular carcinoma. AB - Salmonella abscess of a tumor is extremely rare, only three occurrences having been described to date. An unusual case is presented in which Salmonella infantis septicemia was the presenting symptom of multicentric hepatocellular carcinoma in a previously healthy 67-year-old man. PMID- 9399401 TI - Female sex and the risk of liver cirrhosis. Collaborative Groups for the Study of Liver Diseases in Italy. AB - BACKGROUND: Evidence on gender-related differences in susceptibility to alcohol induced liver diseases is questionable with regard to both methodologic and clinical aspects. With the aim to assess the role of gender in the risk of liver cirrhosis, independently and in combination with known risk factors, data from three case-control studies performed in various Italian areas were analysed. METHODS: The cases were 462 cirrhotic patients (300 men and 162 women) admitted for the first time to hospital for liver decompensation. Controls were 651 patients (355 men and 296 women) admitted to the same hospitals during the same period as the cases, for acute diseases unrelated to alcohol. Alcohol consumption was expressed as lifetime daily alcohol intake. RESULTS: A significant and independent associations between alcohol intake, chronic hepatitis B and C virus infections, and the risk of liver cirrhosis was observed. The effect of alcohol intake was multiplicatively increased in women. The odds ratio (OR) increased from 1.0 (reference category: men, lifetime abstainers) to 31.4 (95% confidence interval (CI), 10.3-95.8) in men drinking more than 100 g/day of alcohol, and from 2.2 (95% CI, 1.0-7.1) in abstaining women to 44.8 (95% CI, 8.2-224.0) in women drinking more than 100 g/day of alcohol. An increased risk of liver cirrhosis associated with female gender independently of alcohol consumption and virus infection was also observed. CONCLUSIONS: A higher susceptibility to alcohol-induced liver diseases was confirmed for women, and an independent effect of female sex on the risk of cirrhosis was observed. Besides alcohol and viruses, some unknown gender-related factor might then be involved in the occurrence of the disease. PMID- 9399403 TI - Helicobacter pylori and non-ulcer dyspepsia. PMID- 9399404 TI - Biochemistry and pharmacology of low molecular weight heparin. PMID- 9399405 TI - Low molecular weight heparins: prophylaxis of venous thromboembolism in surgical patients. PMID- 9399406 TI - Comparison of the relative efficacy and safety of low molecular weight heparin and unfractionated heparin for the treatment of deep venous thrombosis. PMID- 9399407 TI - Treatment of arterial thrombosis with low molecular weight heparins. PMID- 9399408 TI - Low molecular weight heparins: practical considerations. PMID- 9399409 TI - Low molecular weight heparins and antithrombotic therapy: results of an audience interactive symposium. PMID- 9399410 TI - Assembly of the dorsal horn somatotopic map. AB - We hypothesize: (a) peripheral innervation densities determine map scales in dorsal horn, (b) dorsal horn cell (DHC) receptive field (RF) geometries are determined by map scales, and (c) morphologies of primary afferents (PAs) and DHCs reflect their developmental history. We suggest the following sequence: (A) PAs project in a somatotopic mediolateral sequence. (B) DHCs assemble prototype RFs by sampling presynaptic neuropil with their dendrites. (C) PAs then project to all levels where their RFs are contained within prototype RFs of DHCs. (D) A competitive mechanism produces the adult form of DHC RFs. (E) Adult distributions of PA terminals and DHC dendrites reflect this developmental history. (F) Mediolateral somatotopic gradients are determined by RF densities of axons entering at the same levels. (G) Map scales at different rostrocaudal levels are determined by somatotopic gradients. (H) Geometries of DHC RFs are determined by constant convergence and divergence of monosynaptic connections. (I) Secondary processes further modify geometries of DHC RFs. (J) Residual self-organizing capacity supports maintenance and plastic mechanisms. We adduce the following evidence: (1) agreement between monosynaptically coupled inputs and cells' excitatory low threshold mechanoreceptive fields; (2) the temporal sequence of events during penetration of the gray matter by PAs; (3)variation of PA terminal and DHC dendritic domains as a function of map scale; (4) somatotopic gradients and geometries of DHC RFs in adult dorsal horn; (5) calculations of peripheral innervation densities and dorsal horn map scales; and (6) constant divergence and convergence between PAs and DHCs. PMID- 9399411 TI - Spatial summation of perceived pressure, sharpness and mechanically evoked cutaneous pain. AB - Psychophysically, spatial summation can be demonstrated as a decrease in threshold accompanying an increased field of stimulation. The present study examined to what extent different mechanically evoked percepts (pressure, sharpness, and pain) show spatial summation. Various probes were used to apply prescribed forces to the dorsal surface of the digits of 19 healthy subjects. The threshold for three perceptual qualities showed differing degrees of spatial summation: sharpness showed no statistically significant spatial summation; pain demonstrated some significant summation (46% on average); pressure showed the greatest degree of spatial summation (76% on average). The lack of significant spatial summation for sharpness threshold is consistent with the theory that perceived sharpness can be evoked by near threshold activity of a single nociceptor. The modest amount of spatial summation for pain implies that distinctly suprathreshold activation of nociceptors is required for mechanically evoked pain perception, and such input summates centrally, but not completely. The greater spatial summation observed for pressure vs. pain thresholds implies a greater degree of central summation for slowly adapting mechanoreceptors vs. nociceptors. PMID- 9399412 TI - Comparative psychophysical characteristics of cutaneous CO2 laser and contact heat stimulation. AB - Psychophysical visual analog scaling can be used to reveal critical determinants of the neural processing underlying non-painful and painful heat sensations produced by radiant and contact heat stimulation. This study determined the stimulus-response (S-R) functions of cutaneous non-painful and painful heat stimuli delivered by an infra-red CO2 laser or by a contact thermode in a series of experiments in healthy volunteers. In experiments 1 (n = 12), with the rating scale anchored at pain threshold, the S-R curve for brief (60 ms) laser pulse stimulation with a beam diameter of 10 mm was a negatively accelerating function. Transformation of laser stimulus intensity (W) into temperatures (degree C) did not change the form of the S-R curve. In experiment 2 (n = 9), using the same laser stimulus parameters as in experiment 1, but without an anchored rating scale, the form of the S-R relationship did not change. In experiment 3 (n = 9), increases of the laser pulse duration up to 5 s and the beam diameter up to 18 mm produced linear S-R curves. In contrast, in experiment 4 (n = 21), the S-R curve for cutaneous contact heat stimuli applied for 5 s with an 18 mm diameter probe was best described by a positively accelerating power function with an exponent greater than 2.0. These experiments have (1) characterized the S-R functions for different parameters of infra-red laser stimulation of the skin, and (2) have shown that the form of the S-R function for innocuous and noxious heat sensation is influenced strongly by the physical conditions of heat stimulus application, including mechanical contact with the skin. PMID- 9399413 TI - Spatial summation of heat induced pain within and between dermatomes. AB - The aim was to study spatial summation within and between ipsi- and contralateral dermatomes at different painful temperatures. For heat stimulation we used a computer controlled thermofoil based thermode. The thermode area could be varied in five discrete steps from 3.14 to 15.70 cm2. When we applied the stimuli within a dermatome, the mean heat pain threshold decreased significantly from 45.6 to 43.5 degrees C as the area was increased from minimum (3.14 cm2) to maximum (15.70 cm2). When the areas were increased involving different dermatomes (both ipsi- or contralateral), we found similar decreases in pain threshold. Spatial summation was also found within and between dermatomes at supra-threshold temperatures (46, 48, 50 degrees C). The study shows that spatial summation of pain is most likely a mechanism acting across segments and is existing from pain threshold to tolerance. PMID- 9399414 TI - Mastication-related neurons in the orofacial first somatosensory cortex of awake cats. AB - In the orofacial area of the first somatosensory cortex (SI), we recorded single unit activity from 699 neurons in 11 awake cats. Fifty-two percent (362/699) were mastication-related neurons (MRNs) showing activity related to some aspects of masticatory movements. MRNs were divided into three types by their activity patterns: (1) the rhythmical type, showing rhythmical bursts in pace with the masticatory rhythm; (2) the sustained type, showing a sustained firing during the period of taking food and (3) the transient (biting) type, showing intense discharges in coincidence with biting hard food. MRNs had mechanoreceptive fields in the perioral, tongue, periodontal and mandibular regions. The activities of perioral rhythmical-MRNs, mandibular transient-MRNs, tongue rhythmical-MRNs and periodontal transient-MRNs were correlated with food texture, while perioral rhythmical-MRNs, perioral sustained-MRNs and tongue sustained-MRMs were not. Both facial and intraoral MRNs were scattered throughout the facial and intraoral projection areas in SI. These findings provide evidence that the orofacial SI monitors masticatory movements for food ingestion. PMID- 9399415 TI - Activation of a wide-spread network of inhibitory neurons in barrel cortex. AB - The one-to-one correspondence of whiskers to barrels in layer IV of rodent somatosensory cortex can be demonstrated by a precise match between columns of heavy 2-deoxyglucose (2DG) label in layer IV barrels and other layers which correspond to stimulated whiskers. While there is specificity of peripheral-to central mapping, the extent to which integration and/or modulation are generated by circuitry within or interactions between the barrel-defined whisker columns is not clear. Following stimulation of selected whiskers, large cells at the layer IV-V boundary throughout the barrel field are heavily labeled by 2-deoxyglucose (2DG) at high resolution. Many of these cells are outside the barrel columns of the stimulated whiskers. Further, the number of cells labeled is not directly related to the number of activated barrel columns. These neurons are not labeled in animals anesthetized before 2DG injection and are not as heavily labeled in barrel fields of somnolent animals. Most of the heavily labeled neurons immunolabel for glutamate decarboxylase (GAD) and are presumed to be inhibitory, while a smaller number of labeled neurons, presumed to be excitatory, immunolabel for glutamate (Glu). Similar populations of large, heavily 2DG-labeled neurons are found in other cortical areas. These relatively few neurons are exceptionally active and may modulate integrative functions of cerebral cortex. PMID- 9399416 TI - Benzene--a review of the literature from a health effects perspective. AB - A literature review of the impact on human health of exposure to benzene was conducted. Special emphasis in this report is given to the health effects reported in excess of national norms by participants in the Benzene Subregistry of the National Exposure Registry--people having documented exposure to benzene through the use of benzene-contaminated water for domestic purposes. The health effects reported in excess (p < or = .01) by some or all of the sex and age groups studied were diabetes, kidney disease, respiratory allergies, skin rashes, and urinary tract disorders; anemia was also increased for females, but not significantly so. PMID- 9399417 TI - Reproductive and neurobehavioral effects of chlorpropham administered to mice in the diet. AB - Chlorpropham was given in the diet to provide levels of 0(control), 0.15, 0.30, and 0.60%, from 5 weeks of age of the F0 generation to 9 weeks of age of the F1 generation in mice, and selected reproductive and neurobehavioral parameters were measured. There were no adverse effects of chlorpropham on either litter size or litter weight and sex ratio at birth. The average body weight of offspring was significantly affected in the low-dose group of each sex and in the high-dose group of females, and was increased in the middle-dose group of males during the lactation period. In neurobehavioral parameters, surface righting at postnatal day (PND) 7 was significantly affected in male offspring in a dose-related manner. Swimming head angle at PND 4 was significantly restrained in male offspring in a dose-related manner, and olfactory orientation at PND 14 was significantly depressed in female offspring in a dose-related manner. The dose levels of chlorpropham in the present study produced some adverse effects in reproductive and neurobehavioral parameters in mice. PMID- 9399418 TI - The effects of carcinogenic methylcholanthrene on carbohydrate residues of NK cells. AB - The present study examines the effect of methylcholanthrene (MCA), a a carcinogenic polycyclic hydrocarbon, on the carbohydrate receptor determinants (RD) on natural killer (NK) cell surface using the bead-coupled lectin assay. Murine NK cells exhibited different degrees of preferential binding to the specific lectins tested. Of the ten lectins tested, five exhibited a positive binding affinity while the remaining five exhibited no or insignificant binding. NK cells bind to beads derivatized with mannose specific lectins: Concanavalin A (Con A), Lens culinaris, and Pisum sativum. NK cells also bind to other lectin beads such as Triticum vulgaris (GalNac) and Vicia villosa (D-GlcNAc). All these lectin beads exhibited greater than 90% adhesion. The underivatized control beads exhibited no NK binding. The NK cells that were exposed to MCA for 2 h demonstrated a significant decrease in lectin bead-cell coupling in a dose dependent manner. MCA (10 micrograms/mL) caused a 17.8%, 40% and 4.7% decrease in binding affinity when introduced to the mannose specific lectins; Con A, L. culinaris and P. sativum beads, respectively. The binding of T. vulgaris and V. villosa to NK cells was inhibited (23.4% and 28%) by MCA treatment. An increase in the dose to 20 micrograms/mL resulted in a greater inhibition in binding affinity towards lectin beads. Con A, 35.3%, L. culinaris, 62.6%, P. sativum, 30.9%, T. vulgaris, 44.2% and V. villosa, 46.2%. The effect of MCA activation and cytotoxic response. Hydrolysis of PI metabolites (PIP and PIP2) cause generation of secondary messenger: inositol-1,4,5-triphosphate and diacylglycerol, both of which elicit an immune response through their products (Ca2+ and PKC) respectively. Identification of the relationship between receptor level, induction of second messenger and cytotoxic activity may resolve the molecular basis of suppression of NK cytotoxicity by MCA and other PAH compounds. PMID- 9399419 TI - Reactive airway disease in patients with prolonged exposure to industrial solvents. AB - This study examines the effects of volatile organic compounds (VOCs) at the workplace on pulmonary function tests. Forty-two patients with a history of industrial exposure to organic solvents and pulmonary symptomatology were studied. Lung function tests were determined utilizing screening spirometry lung volumes, diffusion capacity and methacholine stimulation test. While only 10-15% of the symptomatic patients had abnormal screening spirometry, 42% of the patients had significantly abnormal methacholine stimulation tests. These data show that exposure to volatile organic solvents is associated with bronchial hyperreactivity not commonly detected by screening spirometry and requires methacholine stimulation testing in individuals with unexplained symptomatology and history of exposure to industrial solvents. These findings indicate that the incidence of bronchial hyperreactivity is underestimated in patients with (1) verifiable exposure history to volatile organic compounds known to be pulmonary irritants, (2) pulmonary symptomatology with normal screening spirometry. PMID- 9399420 TI - Determination of the distribution of malathion in rats following various routes of administration by whole-body electronic autoradiography. AB - The distribution of [14C]-malathion, an organophosphorus pesticide, in rats after intravenous, oral and dermal administration was carried out using electronic autoradiography of whole body sections of treated animals. The study indicated that a major difference in the disposition of [14C]-malathion occurred following various routes of administration to rats. Following intravenous administration, the liver and kidney accumulated extremely high levels of the chemical. After oral administration, [14C]-malathion absorption from the stomach was slow and its excretion followed mostly the fecal route. Dermal application of [14C]-malathion may represent a high risk for exposure to the organophosphorus pesticide where the entire skin, not only the site of application, may act as reservoir for the compound. PMID- 9399421 TI - Dioxin and dioxin-like compounds in soil, Part I: ATSDR interim policy guideline. Agency for Toxic Substances and Disease Registry. PMID- 9399423 TI - Development of birth weight in Austria from 1970-1995. AB - We analysed the development of birth weight of all Austrian live births in the period 1970-1995. The relationship between birth weight and the variables maternal age, length and sex of the newborn and year of birth is described by means of a linear regression model. Over the 26 years an increase of up to 60 g was observed in the mean birth weight. This positive trend may partly be due to the extensive use of the maternity care program ("mother-and-child Passport") introduced in 1974. Maternity payment was made to the mother it she underwent at least five examinations within the antenatal care program. This payment was reduced from 15,000 AS in 1996 to 2000 AS as from January 1st 1997, a measure which may lead to a reduction in the use of this maternity care program, with consequent negative implications for the morbidity of both mother and child and for the positive trend of birth weight development over the past decades. PMID- 9399422 TI - Dioxin and dioxin-like compounds in soil, Part II: Technical support document for ATSDR interim policy guideline. PMID- 9399424 TI - Lipaemia and liver composition in pregnant rats consuming olive oil and olive oil used for frying. AB - The effect of the consumption of unused olive oil (polar content, 2%; oleic acid, 78.9 mg/100 mg oil, and linoleic acid 7 mg/100 mg oil) and olive oil used discontinuously for frying potatoes 15 times (polar content, 9%; oleic acid, 75.8 mg/100 mg oil and linoleic acid 6.2 mg/100 mg oil) was studied in pregnant rats with the aim of better understanding the relationship between the consumption of fat used in frying and lipid metabolism during periods of intense anabolism. Trials were performed in pregnant Wistar rats, divided into 2 groups and fed isocaloric diets in which the fat content (15% wt/wt) consisted of unused olive oil (P1) or oil previously used for frying (P2), and the results were compared with those of nonpregnant rats fed unused olive oil (NP1) and olive oil used for frying (NP2). Pregnancy increased (p < 0.01) food intake, body weight, weight gain, and food efficiency ratio (P2 vs NP2 and P1 vs NP1, respectively), but the treatment of oil included in the diets did not alter these parameters. Gestation significantly increased the serum triglyceride (TG) (p < 0.01) and total cholesterol (TC) (p < 0.05) concentrations and diminished that of phospholipids (PH) (p < 0.01). A significant effect of the type of oil consumed and a pregnancy x oil interaction on Tg and PH levels was observed. The weight of the liver and its fat content increased significantly (p < 0.05) as a result of pregnancy. Liver TC, TG, and PH increased (approximately 3 times the original values) during gestation, but no significant differences due to the intake of used or unused oil (P2 vs P1) were observed. The results indicate that the consumption of moderately altered olive oil, as the sole source of fat, does not alter the effect of pregnancy on the mothers' weight gain, lipaemia, and hepatic fat composition to any important degree. PMID- 9399425 TI - Obesity as a major determinant of underreporting in a self-administered food frequency questionnaire: results from the EPIC-Potsdam Study. AB - The phenomenon of underreporting of dietary intake has been observed previously in many epidemiologic studies. In this study it was investigated whether dependencies exist between energy intake obtained by a semi-quantitative, self administered food frequency questionnaire and lifestyle or anthropometric factors, particularly obesity. The study population consisted of 2,531 subjects, men aged 40 to 64 years and women aged 35 to 64 years from the general population of Potsdam and the surrounding areas. First, subjects were allocated into quintiles of the ratio 'reported energy intake (EI)' to 'calculated basal metabolic rate (BMR)' as a measure of age and weight adjusted energy intake. No apparent dependencies between socio-economic variables and the ratio EI/BMR were observed. Among anthropometric variables, BMI and related measures of obesity were inversely related to the ratio EI/BMR in men and women. While dietary intake was directly related to the ratio EI/BMR in absolute quantities, energy adjusted intake of fat, protein, carbohydrate, and alcohol was found to be independent of this ratio. Energy adjusted food group consumption was also found to be independent of the ratio EI/BMR, showing only slightly increasing trends across quintiles of EI/BMR for cereals and fats, and a slightly decreasing trend for sweet foods in women. When subjects were classified into three categories of BMI, reported energy intake decreased across categories. Estimated energy expenditure based on BMR was increasing with BMI categories. A close direct relationship was observed between BMI categories and the difference between reported energy intake and estimated energy expenditure. It is concluded that obesity is a major determinant of under-reporting. Energy adjusted dietary variables were found to be largely independent of such methodological influences. PMID- 9399426 TI - Exercise-induced sweat nitrogen excretion: evaluation of a regional collection method using gauze pads. AB - The exercise-induced sweat nitrogen excretion was investigated during a 45-minute run at moderate intensity on a treadmill. Sweat was collected with a regional collection technique using gauze pads and compared with the whole-body wash-down (WBW) method. In the regional collection, sweat was sampled from the upper back (UB), lower back (LB), abdomen (AB), and thigh (TH). Additionally, the relation of sweat urea, ammonia, and amino acids was investigated with the regional collection method during a second 45-minute run. Independent of the sweat collection method, a significant and positive correlation was found between sweat rate and the excretion rate of the largest nitrogen fraction urea, suggesting that the sweating response to exercise might be one of the most important factors determining absolute sweat nitrogen losses. The urea nitrogen excretion was nearly 140 mg.h-1 in the second run, representing the largest nitrogen fraction. Ammonia nitrogen and amino acid-derived nitrogen rate were approximately 30 mg.h 1 and 10 mg.h-1, respectively. The comparison of the sampling methods during the first run revealed that the urea nitrogen rate was significantly higher, but the ammonia nitrogen rate significantly lower in the WBW. After summing urea and ammonia nitrogen, no significant difference between the methods was observed anymore, except for UB. It is concluded that the regional collection method using gauze pads is a valuable approach to measure exercise-induced sweat nitrogen losses during moderate running exercise. PMID- 9399427 TI - [Osteoid osteoma of the hand]. PMID- 9399428 TI - A study on biliary ductal system and bile fistula in the American alligator, Alligator mississippiensis. AB - The anomalous arrangement of bile ducts in the Crocodylia has not been fully appreciated. A clear understanding of biliary anatomy is necessary in order to create complete bile drainage in these reptiles. The object of this study was to clarify the anatomy of the bile ductal system and to establish total bile fistulas in the American alligator, Alligator mississippiensis. Bile duct anatomy was studied in 104 juvenile alligators, and bile fistulas were constructed in seven alligators. In 93 out of 104 (89%) of the juveniles dissected there was an interconnection between the right and left hepatic duct before the right hepatic duct emptied into the gallbladder. The left hepatic duct then entered the duodenum independently of the cystic duct which drained the gallbladder directly into the duodenum. In 8% of the animals, the left hepatic duct did not enter the duodenum but joined with the right duct, forming a common hepatic duct that emptied into the gallbladder. In 3% of the cases, the right hepatic duct emptied into the gallbladder, while the left duct had no communication with the right hepatic duct and drained separately into the duodenum. This arrangement of bile ducts is similar to that seen in birds and reflects the common ancestry of crocodiles and birds. In other reptiles, the biliary system shows much more variability and is different from the alligator. In five of seven alligators in which total biliary diversion was attempted, the biliary catheter remained in place and stayed patent from 2-7 weeks. Bile flow was extremely low (1.5-2.5 ml/24 h) when compared to that of mammals (80-100 ml/24 h). This study demonstrates the variable nature of the biliary ductal system in Alligator mississippiensis and suggest a method for constructing an effective total bile fistula in these animals. PMID- 9399429 TI - Abnormal limb regeneration in the short-toes mutant of the axolotl, Ambystoma mexicanum: studies of age, level of amputation, and extracellular matrix. AB - Limb regeneration in the short-toes axolotl is impaired. Our goal was to characterize the regeneration process in this mutant by histological and immunocytochemical methods. Previous research indicates that age and a defective basement membrane may be instrumental factors in short-toes axolotl regeneration (Del Rio-Tsonis et al. [1992] Proc. Natl. Acad. Sci. U.S.A., 89:5502-5506). The present results show that limb regeneration can occur even in older (1-2-year old) short-toes axolotls. The process was always significantly delayed, but the time required for complete regeneration varied. Even so, the basement membrane of short-toes regenerates showed no differences in thickness or shape compared with wild-type regenerates. Distally amputated short-toes limbs gave rise to more digits in the regenerate, indicating that regeneration may be somewhat dependent on the level of amputation. Since extracellular matrix (ECM) remodeling occurs extensively during regeneration, we compared the ECM of the short-toes and wild type regenerates using monoclonal antibodies (mAbs) MT2 and ST1 (Tassava et al. [1996] Wound Rep. Reg., 4:75-81). The short-toes regenerates showed decreased reactivity to mAb MT2, which identifies type XII collagen, an ECM protein that is normally unregulated during regeneration, and increased reactivity to mAb ST1, which identifies a limb ECM component that typically undergoes breakdown in the distal stump. Thus, impaired regeneration in the short-toes axolotl is correlated with impaired ECM remodeling in the distal limb stump. This supports the view that ECM remodeling plays an important role in regeneration. PMID- 9399430 TI - Hormonal induction of thumb pads and the evolution of secondary sexual characteristics of the Southeast Asian fanged frog, Rana blythii. AB - The fanged frogs of Southeast Asia do not express most of the hormone-dependent secondary sexual characteristics such as thumb pads that are common to other ranid frogs. At the same point in the evolutionary history of the group that these androgen-mediated characteristics are lost, male parental care first evolves. This behavior is often correlated with low androgen levels. Prior work indicates that in one of the fanged frogs, Rana blythii, adult males have low androgen levels compared to North Temperate species of Rana. This leads to the question of whether these low androgen levels are related to the unusual male parental care and the lack of expression of the thumb pad and other hormone dependent secondary sexual characteristics in this species. We tested that hypothesis by examining the effects of exogenous dihydrotestosterone supplements on the expression of thumb pads in Rana blythii. Dihydrotestosterone injections appear to stimulate the expression of the thumb pad in R. blythii. These results support the hypothesis that low androgen levels are involved in the loss of the thumb pad in R. blythii. This work provides an example of how mapping characters on phylogenies can be a powerful approach for gaining insights into proximate physiological mechanisms of selection at the evolutionary level. PMID- 9399431 TI - Characterization of vitellogenin and vitellin from land crab Potamon potamios: identification of a precursor polypeptide in the molecule. AB - A female-specific protein, vitellogenin (Vg), and its corresponding egg vitellin (Vn) were identified in the land crab Potamon potamios. Electrophoretic analysis of the hemolymph protein during the annual reproduction cycle revealed that the processing of vitellogenesis did not occur in males or in females of a previtellogenic stage. The analysis of ovarian and egg extracts revealed the presence of Vn, which was identical to Vg. Both Vg and Vn were present in three different aggregational states that represented monomeric, dimeric, and trimeric species of the proteins. In both proteins, the predominant form was the dimeric, which had a molecular mass of 551 kDa for Vg and 510 kDa for Vn. Under denaturing conditions, each of the individual Vg and Vn aggregates released three polypeptides with molecular masses of 115, 105, and 85 kDa. In spite of the difference in terms of native molecular mass, the Vg and Vn were similar in their lipid, carotenoid, and carbohydrate composition. However, Vg of some animals contained a fourth polypeptide with a molecular mass of 181 kDa. This polypeptide and the three other Vg and Vn polypeptides were immunoreactive against anti-Vg prepared from the 85 kDa Vg polypeptide. Furthermore, proteolytic cleavage experiments confirmed that the 115 and 105 kDa Vg polypeptides were derived from the 181 kDa polypeptide. We conclude that the presence of the 181 kDa polypeptide in the Vg molecule resulted in the higher molecular mass of Vg. PMID- 9399432 TI - Duration of the luteotrophic memory of the stud male odors formed in the female mouse. AB - Presence of the stud male prevents the novel male-induced block to early pregnancy in mice, while reexposure to the stud male after the pregnancy block induces pseudopregnancy in the females. These two phenomena involve luteotrophy in the female, and a (luteotrophic) memory of the stud male characteristics formed in her is necessary for such a luteotrophic response. Two experiments were conducted to evaluate the effective duration of the "luteotrophic memory" of the stud male odors formed in the female mouse. The first experiment investigated whether exposure to excreta of the stud male at different intervals will induce pseudopregnancy in pregnancy-blocked females; some females mated with a second male and experienced a second consecutive pregnancy failure before the exposure to the excreta of the first stud male. The females, mated once or twice, failed to exhibit pseudopregnancy when exposure to the excreta of the first stud male was begun 8 or 9 days after the first mating. In the second experiment, each pregnancy-blocked female was mated with the (alien) male that induced failure of the first pregnancy and then housed with simultaneous access to the excreta of another alien male and that of the first stud male. Excreta of the first stud male did not prevent failure of the second pregnancy. The results indicate that the (luteotrophic) memory of the stud male-originating olfactory cues formed in the female or its effectiveness in reactivating the luteotrophic system (when the female is reexposed to the odors of the first stud male) lasts only up to about day 7 postcoitum. The present results also show that exposure to only the excreta of the stud male can prevent the pheromonal block to pregnancy or induce pseudopregnancy after pregnancy block in the female mouse. PMID- 9399433 TI - Regulation of levels of proline as an osmolyte in plants under water stress. AB - Compatible osmolytes are potent osmoprotectants that play a role in counteracting the effects of osmotic stress. Proline (Pro) is one of the most common compatible osmolytes in water-stressed plants. The accumulation of Pro in dehydrated plants is caused both by activation of the biosynthesis of Pro and by inactivation of the degradation of Pro. In plants, L-Pro is synthesized from L-glutamic acid (L Glu) via delta(1)-pyrroline-5-carboxylate (P5C) by two enzymes, P5C synthetase (P5CS) and P5C reductase (P5CR). L-Pro is metabolized to L-Glu via P5C by two enzymes, proline dehydrogenase (oxidase) (ProDH; EC 1.5.99.8) and P5C dehydrogenase (P5CDH; EC 1.5.1.12). Such metabolism of Pro is inhibited when Pro accumulates during dehydration and it is activated when rehydration occurs. Under dehydration conditions, when expression of the gene for P5CS is strongly induced, expression of the gene for ProDH is inhibited. By contrast, under rehydration conditions, when the expression of the gene for ProDH is strongly induced, the expression of the gene for P5CS is inhibited. Thus, P5CS, which acts during the biosynthesis of Pro, and ProDH, which acts during the metabolism of Pro, appear to be the rate-limiting factors under water stress. Therefore, it is suggested that levels of Pro are regulated at the level of transcriptional the genes of these two enzymes during dehydration and rehydration. Moreover, it has been demonstrated that Pro acts as an osmoprotectant and that overproduction of Pro results in increased tolerance to osmotic stress of transgenic tobacco plants. Genetically engineered crop plants that overproduce Pro might, thus, acquire osmotolerance, namely, the ability to tolerate environmental stresses such as drought and high salinity. PMID- 9399434 TI - Expression and internal feedback regulation of ACC synthase and ACC oxidase genes in ripening tomato fruit. AB - We have examined whether or not a positive feedback regulation of gene expression for 1-aminocyclopropane-1-carboxylate (ACC) synthase and ACC oxidase also operates in ripening tomato (Lycopersicon esculentum) fruit during the burst of ethylene production. Two cDNA fragments for ACC synthase and one for ACC oxidase were cloned with high homology to already known genes involved in ethylene biosynthesis in ripening tomato fruit. Accumulation of mRNAs which hybridize to these cDNA probes were induced in mature green fruit within two days by treatment with propylene. In the fruit ripened from the turning stage, red color development, ethylene production, ACC content, and activities of ACC synthase and ACC oxidase increased as maturity progressed. The abundance of two ACC synthase and one ACC oxidase mRNAs in the fruit increased from the turning to pink stage and were followed by a slight decline towards the red stage. These increases in mRNAs abundance with ripening were prevented to a large extent by treatment with the ethylene action inhibitor, 1-methylcyclopropene (MCP). This was most pronounced in the fruit treated with MCP at the turning stage, in which the accumulation of ACC synthase and ACC oxidase transcripts was almost completely eliminated in the first two d, precisely the same stage at which the control fruit had the greatest level of each mRNA accumulation. The inhibition of transcript accumulation recovered to the control level within two to four d. MCP also decreased ethylene biosynthetic activity, although this decrease did not reflect the reduction in the mRNAs accumulation. These results suggest that a strong positive feedback regulation is involved in ethylene biosynthesis at the gene transcriptional level in tomato fruit, even at the stage with a burst of ethylene production. PMID- 9399435 TI - Generation of superoxide anion and localization of CuZn-superoxide dismutase in the vascular tissue of spinach hypocotyls: their association with lignification. AB - The sites of generations of superoxide anions and hydrogen peroxide in cross sections of hypocotyls from spinach seedlings were located by staining with nitroblue tetrazolium (NBT) and with starch-iodide, respectively. Formazan, produced upon the reduction of NBT by superoxide, was observed mainly in the vascular tissue only in the presence of inhibitors of CuZn-superoxide dismutase (CuZn-SOD), and its formation was suppressed under anaerobic conditions. Thus, NBT was reduced to formazan specifically by the superoxide anions generated in vascular tissue. The reduction of NBT was suppressed by inhibitors of NAD(P)H oxidase, but neither by cyanide nor azide, indicating the involvement of NAD(P)H oxidase in the generation of superoxide anions in the vascular tissue. Starch-I2 complex also was formed in the vascular tissue, but not in the presence of either the CuZn-SOD inhibitor or the NAD(P)H oxidase inhibitor, indicating that the hydrogen peroxide is produced via the catalytic disproportionation with CuZn-SOD of the superoxide generated by NAD(P)H oxidase. Generations of superoxide anions and hydrogen peroxide in the vascular tissue were particularly apparent in the xylem and associated with the sites of distribution of CuZn-SOD as determined by an immunohistochemical method, and also with the location of lignin as determined by the phloroglucin-HCl reaction. PMID- 9399436 TI - Morphological responses and molecular modifications in tomato plants after mechanical stimulation. AB - Study of the growth responses of Lycopersicon esculentum (Mill. cv. VFN8) to mechanical stimulation applied to a single young internode showed a rapid and sharp decrease in stem elongation and an inhibition of elongation of several internodes, indicative of information transmission in the plant. A new tomato cDNA partial clone encoding calmodulin was isolated and used to study the time course of the gene induction in response to the rubbing treatment. Northern blot analysis showed a maximum accumulation of calmodulin mRNA 2 h after mechanical stimulation, not only in the rubbed internode, but also in upper and lower internodes and in young leaves. Treatment of the plant with calcium and EGTA showed the involvement of calcium and, in particular, intracellular calcium in calmodulin gene expression and cellular response. PMID- 9399437 TI - Gibberellin A3 causes a decrease in the accumulation of mRNA for ACC oxidase and in the activity of the enzyme in azuki bean (Vigna angularis) epicotyls. AB - Differential screening, aimed at the isolation of cDNA clones of mRNAs whose accumulation is influenced by GA3, resulted in the isolation of a cDNA clone of an mRNA whose level was decreased by GA3 in segments of epicotyls of Vigna angularis. The putative protein encoded by this cDNA resembled the 1 aminocyclopropane-1-carboxylate oxidases (ACC oxidases) identified in other plant species (about 80% homology at the amino acid level). Thus, the corresponding gene was designated AB-ACO1 (azuki bean ACC oxidase). GA3 also decreased the activity of ACC oxidase in azuki bean epicotyls, but it did not decrease the rate of ethylene evolution. In fact, GA3 increased the rate of ethylene evolution and the level of ACC. Thus, GA3 seemed to increase the production of ethylene by promoting the synthesis of ACC. PMID- 9399438 TI - The requirements for Ca2+, protein phosphorylation, and dephosphorylation for ethylene signal transduction in Pisum sativum L. AB - The role of Ca2+ and protein phosphorylation in the transduction of the ethylene signal resulting in induction of 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase has been studied in peas (Pisum sativum L.) by a pharmacological approach. 2,5-Norbornadiene (NBD) and aminoethoxyvinylglycine (AVG) reduced the basal level of ACC oxidase transcript and its enzyme activity. Only NBD was shown to inhibit the ethylene response, the accumulation of ACC oxidase transcript and the stimulation of its enzyme activity. Ethylene influenced 45Ca2+ influx into the segment tissues from pea epicotyls, and ethylene glycol-bis(beta-aminoethyl ether)N,N,N'N'-tetraacetic acid (EGTA) a Ca2+ chelator, inhibited the ethylene response. Ca2+ depletion by pretreatment with EGTA also blocked the ethylene response, which almost completely recovered when Ca2+ was added exogenously after Ca2+ depletion. Ca2+ channel blockers, verapamil, and LaCl3, used to certify the role of extracellular Ca2+, all inhibited the ethylene response. A protein kinase inhibitor, 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine (H-7), and protein phosphatase inhibitors, vanadate and okadaic acid, also inhibited the ethylene response. The results of the present study suggest that Ca2+ influx from the extracellular space, protein phosphorylation, and dephosphorylation are required for the induction of ACC oxidase by ethylene. PMID- 9399439 TI - Purification and characterization of arginine decarboxylase from soybean (Glycine max) hypocotyls. AB - Arginine decarboxylase (EC 4.1.1.19) was purified from soybean, Glycine max, hypocotyls by a procedure which includes ammonium sulfate fractionation, acetone precipitation, gel filtration chromatography, and affinity chromatography. Using this procedure, ADC was purified to one band in non-denaturing PAGE. The purified ADC has an M(r) of 240 kDa based on gel filtration chromatography and is a trimer of identical subunits which has an estimated M(r) of 74 kDa based on SDS-PAGE. ADC is active between 30 and 50 degrees C and has a Km value of 46.1 microM. ADC is very sensitive to agmatine or putrescine but not to spermidine or spermine. In the presence of 0.5 mM agmatine (or putrescine), the enzyme activity was inhibited by 70%. However, at the same concentration of spermidine (or spermine), the enzyme activity was inhibited by only 10-20%. PMID- 9399440 TI - Differential expression of ADC mRNA during development and upon acid stress in soybean (Glycine max) hypocotyls. AB - Arginine decarboxylase (ADC) is one of the key enzymes in the biosynthesis of putrescine in plants. The regulation of its activity depends on the physiological condition, developmental stage, and type of tissue. We have cloned ADC cDNA from soybean (Glycine max) hypocotyls to understand the regulation mechanisms of this enzyme activity. Using the cDNA clone, we examined the relationship between changes in the ADC activity and the level of ADC mRNA during development, in different tissues, and upon acid stress. The ADC activity began to increase 2 d after initiation of germination, reached a peak at the 5th d, and then declined. This change in the enzyme activity was preceded by similar changes in the level of the mRNA. The ADC activity was expressed tissue-specifically; this expression was well corelated with the mRNA content of the respective tissues. Incubation of the 5-d-old hypocotyls in pH 3 potassium phosphate solution caused a rapid increase in ADC activity. Within 2 h of acid treatment, the ADC activity increased more than threefold. This increase was also preceded by a corresponding increase in the mRNA content and was also regulated tissue-specifically. These results suggest that the change in the content of ADC mRNA has an important role in the regulation of the enzyme activity during early development, in different tissues, and upon acid stress. PMID- 9399441 TI - Cloning of maize ferredoxin III gene: presence of a unique repetitive nucleotide sequence within an intron found in the 5'-untranslated region. AB - A genomic clone encoding the precursor of a non-photosynthetic ferredoxin (Fd III) from maize has been isolated and characterized. In comparison with the corresponding cDNA, the gene (fedIII) was found to be interrupted in the 5' untranslated region by an intron consisting of 3,037 bp. About 80% of the total region of this intron is organized into four classes of tandemly repeated sequences with monomeric lengths of about 200 bp, 320 bp, 130 bp, and 90 bp. This unique intron organization is present in the fedIII gene of three related maize cultivars examined. PMID- 9399442 TI - Cervical Spine Research Society Presidential Address: a 25-year correlation with history. PMID- 9399443 TI - Murine nucleus pulposus-derived cells secrete interleukins-1-beta, -6, and -10 and granulocyte-macrophage colony-stimulating factor in cell culture. AB - STUDY DESIGN: Cultures established from murine disc-derived cells were stimulated by lipopolysaccharide. The cells' capacity to secrete proinflammatory cytokines and interleukin-10 with and without lipopolysaccharide stimulation was determined using enzyme-linked immunosorbent assays. OBJECTIVES: To determine the capacity of disc-derived cells to secrete proinflammatory cytokines, and the effect of lipopolysaccharide stimulation on such secretion. SUMMARY OF BACKGROUND DATA: The pathophysiology of compressive radiculopathy is unclear. Inflammation is a possible explanation. Proinflammatory cytokine secretion was demonstrated in herniated nucleus pulposus. It is unknown whether these cytokines are secreted from disc-derived cells or from infiltrating inflammatory cells in the herniated nucleus pulposus. METHODS: Discs were microsurgically harvested from inbred mice and cut to allow the nucleus pulposus to establish cell culture. A study group was exposed to lipopolysaccharide stimulation. Media were harvested from the study and control groups 24 hours later. Secretion of interleukins-1-, -6, and 10, granulocyte-macrophage colony-stimulating factor, and tumor necrosis factor alpha were determined using enzyme-linked immunosorbent assays. RESULTS: Basal secretion of interleukins-6 and -10, but no basal secretion of interleukin-1-, granulocyte-macrophage colony-stimulating factor, or tumor necrosis factor-alpha was detected. Secretion of interleukin-1- rose from zero to 27.69 pg/10(5) cells, and granulocyte-macrophage colony-stimulating factor secretion rose from zero to 9.77 pg/10(5) cells after lipopolysaccharide stimulation. A 75-fold increase in interleukin-6 secretion and a 150-fold increase in interleukin-10 secretion were detected after stimulation with lipopolysaccharide. No tumor necrosis factor alpha secretion was detectable. All result had high statistical significance (all P < 0.001). CONCLUSIONS: Cultured murine disc-derived cells have the capacity to secrete proinflammatory cytokines and interleukin-10 in the absence of inflammatory cells. This finding supports the hypothesis that disc-derived cells are capable of initiating or amplifying an inflammatory process. PMID- 9399444 TI - Clinical course of conservatively managed rheumatoid arthritis patients with myelopathy. AB - STUDY DESIGN: The clinical course of rheumatoid arthritis patients with myelopathy who do not undergo surgery was studied. OBJECTIVES: To establish a more accurate prognosis for rheumatoid arthritis patients who do not undergo surgery. SUMMARY OF BACKGROUND DATA: Cervical myelopathy has been reported in two thirds of rheumatoid arthritis patients with atlantoaxial dislocation. Atlantoaxial fusion, or occipitocervical fusion, is widely performed on these patients. However, several researchers reported serious complications from the surgery, including nonunion, worsening myelopathy, and high mortality. The natural course of disease in rheumatoid arthritis patients with myelopathy should be known before definitive treatments can be outlined. MATERIALS AND METHODS: Twenty-one rheumatoid arthritis patients with myelopathy resulting from atlantoaxial dislocation were studied. Fourteen of the 21 cases were associated with upward migration of the odontoid process. All of these patients were recommended for surgery, but they refused. Patients were reviewed by direct examination yearly. Radiographic changes and clinical course, including the survival rate, were observed. RESULTS: Atlantodental interval and Redlund-Johnell measurements deteriorated. The patients showed no neural improvement, and deterioration was found in 16 (76%) cases during follow-up. All patients became bedridden within 3 years of the onset of myelopathy. Seven of the 21 patients died suddenly for unknown reasons, 3 died of pneumonia, and 1 died of multiple organ failure. The three sudden-death cases showed progressive upward migration of the odontoid process. The cumulative probability of survival was 0% in the first 7 years after the onset of myelopathy. CONCLUSIONS: The clinical results for rheumatoid arthritis patients with myelopathy treated without surgery are extremely poor. Surgical treatment is recommended for rheumatoid arthritis patients with myelopathy. PMID- 9399445 TI - Neurologic outcome of early versus late surgery for cervical spinal cord injury. AB - STUDY DESIGN: A prospective analysis evaluating neurologic outcome after early versus late surgery for cervical spinal cord trauma. OBJECTIVES: The study was conducted to determine whether neurologic and functional outcome is improved in traumatic cervical spinal cord-injured patients (C3-T1, American Spinal Injury Association grades A-D) who had early surgery (<72 hours after spinal cord injury) compared with those patients who had late surgery (>5 days after spinal cord injury). SUMMARY OF BACKGROUND DATA: There is considerable controversy as to the appropriate timing of surgical decompression and stabilization for cervical spinal cord trauma. There have been numerous retrospective studies, but no prospective studies, to determine whether neurologic outcome is best after early versus late surgical treatment for cervical spinal cord injury. METHODS: Patients meeting appropriate inclusion criteria were randomized to an early (<72 hours after spinal cord injury) or late (>5 days after spinal cord injury) surgical treatment protocol. The neurologic and functional outcomes were recorded from the acute hospital admission to the most recent follow-up. RESULTS: Comparison of the two groups showed no significant difference in length of acute postoperative intensive care stay, length of inpatient rehabilitation, or improvement in American Spinal Injury Association grade or motor score between early (mean, 1.8 days) versus late (mean, 16.8 days) surgery. CONCLUSIONS: The results of this study reveal no significant neurologic benefit when cervical spinal cord decompression after trauma is performed less than 72 hours after injury (mean, 1.8 days) as opposed to waiting longer than 5 days (mean, 16.8 days). PMID- 9399446 TI - The management of unilateral lateral mass/facet fractures of the subaxial cervical spine: the use of magnetic resonance imaging to predict instability. AB - STUDY DESIGN: Retrospective review of the clinical course and cervical spine plain radiographs, computed tomography, and magnetic resonance imaging of 24 consecutive patients for a 2-year period with a unilateral lateral mass/facet fracture. OBJECTIVE: To propose a treatment algorithm for the management of unilateral lateral mass/facet fractures of the subaxial cervical spine based on ligamentous injury detected by magnetic resonance imaging. SUMMARY OF BACKGROUND DATA: There have been no previous reports of the use of magnetic resonance imaging to predict clinical instability. METHODS: A retrospective review of the clinical course of all unilateral mass/facet fractures identified over a 2-year period was conducted. All cervical spine plain radiographs, computed tomography scans, and magnetic resonance images were reviewed by a neuroradiologist blinded to the clinical course of the patient. Magnetic resonance T1-weighted and inversion recovery images were used to evaluate the integrity of the facet region, interspinous ligament, anterior longitudinal ligament, and posterior longitudinal ligament. RESULTS: Twenty-four unilateral lateral mass/facet fractures were identified. Only six initial cervical spine series demonstrated a bony abnormality at the level of the fracture. The fractures were identified by computed tomography and were almost all nondisplaced or minimally displaced. Less than half of the fractures extended ventrally to involve the transverse process or foramen transversarium or dorsally to involve the lamina. Twelve fractures were nonoperatively treated and 12 were treated surgically for stabilization. Ten patients in the operative group presented with or developed a subluxation. Nine of these patients had injury to at least three of the four ligaments evaluated by magnetic resonance imaging. In the nonoperative group, only three patients had extensive ligamentous injury at the level of the fracture. All three of these patients were lost to follow-up. CONCLUSIONS: Plain radiographs of the cervical spine lack sensitivity to detect the presence of lateral mass/ facet fractures. The appearance of the fracture on computed tomography does not indicate instability. The degree of ligamentous injury at the level of the fracture demonstrated on magnetic resonance imaging correlates with instability in this series. Operative stabilization may be indicated for unilateral lateral mass fractures that present with a subluxation or that have injury to at least three of the following ligaments: the facet region, the interspinous ligament, the anterior longitudinal ligament, and the posterior longitudinal ligament. However, before a definitive management plan can be formulated, results from this small series require further validation. PMID- 9399447 TI - Three-level anterior cervical discectomy and fusion: radiographic and clinical results. AB - STUDY DESIGN: A retrospective study of 16 patients who underwent the modified Robinson anterior cervical discectomy and fusion at three operative levels. OBJECTIVES: To provide long-term follow-up data on the surgical success and patient outcome of three-level anterior cervical discectomies and fusions. SUMMARY OF BACKGROUND DATA: The success of arthrodesis for anterior cervical fusion depends on several factors, including the number of surgical levels. To the authors' knowledge, there are no long-term follow-up reports to describe the arthrodesis rate and outcome for patients having specifically three-level discectomy and fusion procedures. METHODS: Sixteen patients, with an average age of 59 years, were followed for an average of 37 months. All had an anterior discectomy, burring of the endplates, and placement of an autogenous tricortical iliac crest graft at three levels. All patients had follow-up office visits with examinations and radiographs. Radiographic union, postoperative pain relief, and neurologic recovery were evaluated. RESULTS: Only 9 (56%) of the 16 patients went on to achieve solid arthrodesis at all three levels. Of the seven patients with pseudarthrosis, two had severe pain and required revision; two had moderate pain and three no pain. Of the nine with the solid fusion, three had mild pain and six no pain, a statistically significant difference in comparing the two outcomes (P < 0.01). All patients with preoperative motor deficit recovered, but two patients in whom a pseudarthrosis had developed had limited improvement in function until the nonunion was surgically repaired. CONCLUSIONS: A three-level modified Robinson cervical discectomy and fusion results in an unacceptably high rate of pseudarthrosis. Although not all pseudarthroses are painful, these data suggest that those with a successful fusion have a better outcome. It is recommended that these patients undergo additional or alternative measures to achieve arthrodesis consistently. PMID- 9399448 TI - Characterization of the scoliosis that develops after pinealectomy in the chicken and comparison with adolescent idiopathic scoliosis in humans. AB - STUDY DESIGN: The characteristics of the scoliosis that develops after pinealectomy in young chickens were determined from weekly posteroanterior radiographs. These data were compared with similar data collected from human patients with adolescent idiopathic scoliosis. OBJECTIVES: To characterize the scoliosis produced in young chickens after pinealectomy and to compare these characteristics with those seen in human patients with adolescent idiopathic scoliosis. SUMMARY OF BACKGROUND DATA: Although it has been recognized that pinealectomy produces scoliosis in chickens, the characteristics of these curves have never been well described other than by simple visual descriptions. METHODS: The characteristics of the scoliosis produced in chickens after pinealectomy done 3 days after hatching were measured from radiographs taken at weekly intervals. These characteristics were compared with similar data collected from human patients with adolescent idiopathic scoliosis. RESULTS: Similarities included development of single and double curves, degree of curvature, stability of the curve, numbers of vertebrae involved, direction of rotation, and progression characteristics. Differences included wedged vertebrae in the chickens, in conjunction with curve development and increased variability in vertebrae involved. CONCLUSIONS: There are many similarities in the development of scoliosis in young chickens after pinealectomy and in children with adolescent idiopathic scoliosis. The few differences might be related to the different biomechanical properties associated with the spine in the two species. PMID- 9399449 TI - Lumbar spinal canal stenosis examined electrophysiologically in a rat model of chronic cauda equina compression. AB - STUDY DESIGN: A model of chronic cauda equina compression with conductive stress was studied electrophysiologically. OBJECTIVE: To analyze the pathophysiology arising from chronic compression electrophysiologically. SUMMARY OF BACKGROUND DATA: This rat model of cauda equina compression that is chronic, not acute, has been reported elsewhere. METHODS: A stainless steel wire and plate were fastened to the spine at L5 of 8 rats 3 weeks old. One year later, the ascending and descending nerve action potentials were recorded and the conduction velocities (CVs) were measured. Electrophysiologic changes after high-frequency stimulation (HFS) were observed. RESULTS: The waveform of the ascending cauda equina action potential at the cauda equina had three peaks, and that at the conus medullaris had a peak followed by a broad wave. The waveform of the descending nerve action potential had two peaks. The mean ascending and descending CVs of the treated rats were slower (P < 0.001) than those of the control rats. In the control rats, the mean CV and mean amplitude after HFS decreased slightly and returned to normal within 30 seconds, and the waveform was unchanged. In treated rats, the mean CV decreased after HFS but returned to normal within 10 minutes. The mean amplitude decreased after HFS and did not return to normal within 10 minutes. The waveform was unchanged. CONCLUSIONS: Because the differences between treated and control rats in amplitude (and CVs) were greater before HFS than after HFS, we concluded that treated rats had disturbance of the blood flow in vessels around the nerves of the cauda equina with histologic damage. In human patients, such disturbance may be one cause of intermittent claudication. PMID- 9399450 TI - Pathogenesis of tears of the anulus investigated by multiple-level transaxial analysis of the T12-L1 disc. AB - STUDY DESIGN: Three transaxial slices, dividing each disc into four layers of equal thickness, were made in each of 19 T12-L1 discs. Naked-eye and stereoscopic examination was used to record abnormalities of the T12 (superior) surface of the upper slice, opposing surfaces of the central slice, and the L1 (inferior) surface of the lower slice. OBJECTIVES: To characterize and quantify structural abnormalities to determine their incidence and three-dimensional arrangement, and to test the hypotheses 1) that the frequency and location of tears of the anulus are related to age and nucleus condition; and 2) that rim lesions initiate the development of concentric tears. SUMMARY OF BACKGROUND DATA: Most previous studies of disc disease have been based on the examination of single sagittal slices, some on single transaxial slices, and a few have used both. This single slice approach underrecords abnormalities that have not involved the disc center, and may inhibit the interpretation of magnetic resonance imaging and computed tomography images. METHODS: Spines from 19 human cadavers (mean age, 47.4 years; range, 20-79 years) were used. An initial transaxial slice through the center of the T12-L1 disc was followed by cranial and caudal transaxial slices midway between the center and endplate. Soft tissues were then removed to allow examination of the endplate. Abnormalities recorded at each stage were summated for all disc levels. The incidence of the abnormalities in each disc sector was analyzed using the Spearman-Rank correlation coefficient and the Bonferroni correction. RESULTS: With the exception of radiating tears, which most commonly affected the posterior disc, the right anterior quadrant tended to show abnormalities more frequently than the other quadrants. Although concentric tears (in 74%), rim lesions (in 47%), and radiating tears (in 47%) were frequent, no correlations were found between these three types of anulus tear. Concentric tears were present after approximately 10% of the anulus had undergone some delamination. Rim lesions correlated with focal thickening of anulus lamellae. One fifth of radiating tears extended to involve the outer anulus zone. CONCLUSIONS: Neither hypothesis was substantiated. Because lesions of the nucleus and anulus lack uniform shape and are three-dimensionally complex, it is inappropriate to interpret cadaver disc disease on single, mid-disc slices. The three different types of anulus tears appear to evolve independent of age and each other. PMID- 9399451 TI - The effect of fatigue on multijoint kinematics and load sharing during a repetitive lifting test. AB - STUDY DESIGN: A repetitive lifting test in the sagittal plane was performed with a submaximal load at a maximal lifting rate to understand the effects of fatigue on kinematic and kinetic measures of performance. OBJECTIVES: To quantify the effect of fatigue during a highly repetitive lifting task, in terms of lifting force transmitted to the load, joint motion patterns, and internal joint load sharing. SUMMARY OF BACKGROUND DATA: Industrial surveillance and epidemiologic data suggest that repetitive lifting is a risk factor for low back pain. Previous studies examining the effect of fatigue have either been constrained to isolated trunk movement, or have not explored the internal load distribution and potential alteration in the loading patterns. METHODS: Sixteen healthy male subjects performed repetitive lifting in the sagittal plane with a load equal to 25% of their maximal lifting capacity, at a maximal lifting rate. Changes in lifting performance were determined from the power transferred to the box, joint kinematics, and joint kinetics. Data from three cycles at the start and end of the exercise were tested for the effect of fatigue using repeated-measures analysis of variance. RESULTS: Fatigue was documented by a reduction in average lifting force and hip and spine torque generation, whereas internal joint load sharing was relatively unchanged. The fatigue was associated with decreased knee and hip motion, and increased lumbar flexion. Decreased postural stability also was evident. CONCLUSIONS: The significant decrease in postural stability and force generation capability because of the repetitive lifting task indicated a higher risk of injury in the presence of unexpected perturbation. Multijoint coordinated lifting tasks provide a more realistic protocol to study neuromuscular fatigue. PMID- 9399452 TI - The effect of spinal destabilization and instrumentation on lumbar intradiscal pressure: an in vitro biomechanical analysis. AB - STUDY DESIGN: In vitro biomechanical testing was performed in human cadaveric lumbar spines, using pressure needle transducers to analyze the effects of spinal destabilization and instrumentation on lumbar intradiscal pressures. OBJECTIVES: To quantify changes in lumbar intradiscal pressures at three adjacent disc levels under conditions of spinal reconstruction, and to evaluate the possibility of pressure-induced disc pathology secondary to spinal instrumentation. SUMMARY OF BACKGROUND DATA: Lumbar intradiscal pressures under in vivo and in vitro conditions and the use and development of spinal instrumentation have been investigated comprehensively. However, the effects of spinal destabilization and instrumentation on lumbar intradiscal pressure have not been delineated clearly. METHODS: In 11 human cadaveric lumbosacral specimens, specially designed pressure needle transducers quantified intradiscal pressure changes at three adjacent disc levels (L2-L3, proximal; L3-L4, operative; and L4-L5, distal) under four conditions of spinal stability: intact, destabilized, laminar hook and pedicle screw reconstructions. Biomechanical testing was performed under axial compression (0-600 N), anterior flexion (+12.5 degrees) and extension (-12.5 degrees), after which the level of degeneration and disc area (cm2) were quantified. RESULTS: In response to destabilization and instrumentation, proximal disc pressures increased as much as 45%, and operative pressure levels decreased 41-55% (P < 0.05), depending on the instrumentation technique. Linear regression and correlation analyses comparing intradiscal pressure to the grade of disc degeneration were not significant (r = 0.24). CONCLUSIONS: Changes in segmental intradiscal pressure levels occur in response to spinal destabilization and instrumentation (P < 0.05). Intradiscal cyclic pressure differentials drive the metabolic production and exchange of disc substances. Conditions of high or low disc pressure secondary to spinal instrumentation may serve as the impetus for altered metabolic exchange and predispose operative and adjacent levels to disc pathology. PMID- 9399453 TI - Vulnerability of the recurrent laryngeal nerve in the anterior approach to the lower cervical spine. AB - STUDY DESIGN: To perform anatomic dissections and measurements of the recurrent laryngeal nerve between the inferior thyroid artery and superior border of the clavicle (mid-portion) on both sides. OBJECTIVES: To determine quantitatively the differences in course and location between the recurrent laryngeal nerves on both sides and to relate this to the vulnerability of the recurrent laryngeal nerve during an anterior approach to the lower cervical spine. SUMMARY OF BACKGROUND DATA: The midportion of the recurrent laryngeal nerve is usually encountered in the anterior approach to the lower cervical spine, especially on the right side. No quantitative regional anatomy describing the course and location of the mid portion of the recurrent laryngeal nerve is available in the literature. METHODS: Fifteen adult cadavers were used for dissections of the recurrent laryngeal nerve. The length of the recurrent laryngeal nerve between the superior border of the clavicle and the inferior thyroid artery, and the angle of the recurrent laryngeal nerve with respect to sagittal plane, were measured bilaterally. In addition, six cross-sections at C7 were obtained to determine the linear distances between esophagotracheal groove and the recurrent laryngeal nerve. RESULTS: The recurrent laryngeal nerve on the right runs in a superior and medial direction, with an angle of 25.0 degrees +/- 4.7 degrees relative to sagittal plane, compared with 4.7 degrees +/- 3.7 degrees on the left. The length of the recurrent laryngeal nerve between the superior border of the clavicle and the inferior thyroid artery is 23.0 +/- 4.4 mm on the left, and 22.8 +/- 4.3 mm on the right. The recurrent laryngeal nerve lies deep within the esophagotracheal groove on the left, but 6.5 +/- 1.2 mm anterior and 7.3 +/- 0.8 mm lateral to the esophagotracheal groove on the right. CONCLUSIONS: The recurrent laryngeal nerve on the right side is highly vulnerable to injury if ligature of the inferior thyroid vessels is not performed as laterally as possible or if retraction of the midline structures along with the recurrent laryngeal nerve is not performed intermittently. Avoiding injury to the recurrent laryngeal nerve, especially on the right side, is a major consideration during an anterior approach to lower cervical spine. PMID- 9399454 TI - Accuracy of using computed tomography to identify pedicle screw placement in cadaveric human lumbar spine. AB - STUDY DESIGN: Utility of using computed tomography to predict pedicle screw misplacement. OBJECTIVE: This study defines the sensitivity and specificity of predicting pedicle screw placement by experienced clinicians using a CT scan image. SUMMARY OF BACKGROUND DATA: In clinical and research settings, the method most commonly used to evaluate pedicle screws placement has been computed tomography. However, no current literature describes the accuracy of this method of evaluating screw placement. METHOD: Cobalt-chrome and titanium alloy pedicle screws of identical size were placed in six cadaveric human lumbar spine. Wide laminectomy was performed to allow complete visualization of the pedicles. Three consecutive lumbar levels were instrumented in each spine, giving 36 pedicle screw placements to identify. The instrumented spines were imaged, and four orthopaedic spine surgeons and a musculoskeletal radiologist were asked to read the images to identify the accuracy of screw placement within the pedicles. RESULTS: The sensitivity rate of identifying a misplaced screw was 67 +/- 6% for cobalt-chrome screws compared with 86 +/- 5% for titanium screws (P < 0.005). The specificity rates of radiographic diagnosis of misplaced pedicle screws were 66 +/- 10% for cobalt-chrome screws and 88 +/- 8% for titanium screws (P < 0.005). Similarly, a statistically significant difference was found in the sensitivity rates of identifying screws placed correctly in the pedicle: 70 +/- 10% for cobalt-chrome screws versus 89 +/- 8% for titanium screws (P < 0.005). Overall accuracy rates were 68 +/- 7% for cobalt chrome screws versus 87 +/- 3% for titanium screws (P < 0.002). CONCLUSION: Reliance on the computed tomography scan data alone in determining accuracy of pedicle screws can lead to inaccuracies in both clinical and research conditions. PMID- 9399455 TI - Differentiation of submaximal from maximal trunk extension effort: an isokinetic study using a new testing protocol. AB - STUDY DESIGN: An evaluation of the relations between concentric and eccentric contractions of the trunk extensors and extension effort performed at maximal and submaximal levels. OBJECTIVE: To define quantitative parameter(s) derivable from isokinetic dynamometry that may differentiate submaximal from maximal trunk extension moment. SUMMARY OF BACKGROUND DATA: Using various consistency-related parameters, researchers in previous studies have not been able confirm the potential of isokinetic dynamometry for identifying submaximal effort during trunk extension. METHODS: Twenty healthy subjects, 8 women and 12 men without low back pain history, aged 21 to 30 years, took part in this study. Testing consisted of three experimental conditions using four intermittent concentric and eccentric contractions at 20 degrees and 60 degrees/second. The first condition, in which subjects were asked to exert maximal concentric and eccentric effort, served as the baseline. In the second condition, subjects were asked to exert 50% of the force measured in the first condition. In the third condition, subjects repeated the second condition but were asked to exert the best reproducible level of force. RESULTS: The highest differentiating power among the experimental conditions was attributed to the intervelocity difference between the concentric and eccentric contractions (P < 0.0001). CONCLUSIONS: This protocol effectively differentiates submaximal from maximal trunk extension effort in normal subjects. PMID- 9399456 TI - Elective cervical discectomy in California: postoperative in-hospital complications and their risk factors. AB - STUDY DESIGN: A retrospective cohort study of short-term outcomes after elective cervical discectomy in California hospitals. OBJECTIVES: To compare the frequency of elective cervical discectomy across population strata, to determine the frequency of adverse outcomes in the early postoperative period, and to identify risk factors for such outcomes. SUMMARY OF BACKGROUND DATA: Previous cervical discectomy series have been too small to analyze risk factors for early complications, and have originated from centers that may not adequately represent the population. METHODS: Computerized hospital discharge abstracts were obtained from the California Office of Statewide Health Planning and Development. Inclusion and exclusion criteria were applied to identify 10,416 routine discectomies at 257 hospitals in 1990-1991. Several categories of postoperative complications were identified, along with inpatient deaths, early reoperations, and nursing home transfers. Logistic regression was used to estimate the independent effects of patient characteristics on short-term outcomes. RESULTS: After adjustment for age and gender, blacks were 51% and Hispanics were 24% as likely as whites to undergo elective cervical discectomy. Overall, 6.7% of patients had one or more reported postoperative complications: 1.8% had noninfectious surgical complications, 1.8% had infectious complications, 4.0% had other medical complications, and 0.35% had unplanned reoperations before discharge. Fourteen inpatient deaths were reported (0.13%). Congestive heart failure, alcohol/drug abuse, chronic lung disease, previous spine surgery, psychological disorders, and chronic musculoskeletal disorders were independently associated with postoperative complications. Even after adjustment, risk was higher with advancing age, higher among women than among men, and higher after posterior fusion than after discectomy without fusion. CONCLUSIONS: The ethnic disparity in cervical discectomy rates suggests overuse among whites or underuse among minority populations. The complication rates reported here are similar to those synthesized from previous literature, except that the lower incidence of neurologic complications reflects our inability to distinguish preoperative from postoperative deficits. Important comorbidities should be identified and treated, if appropriate, before cervical spine surgery. PMID- 9399457 TI - Lower urinary tract symptoms in lumbar root compression syndromes: a prospective survey. AB - STUDY DESIGN: A prospective, observational survey. OBJECTIVES: To describe lower urinary tract symptoms in uncomplicated lumbar root compression syndromes with special reference to prevalence, nature, and severity, and to analyze whether the occurrence of lower urinary tract symptoms correlates with age, pain, analgesic intake, or the type and level of compression. SUMMARY OF BACKGROUND DATA: Lower urinary tract symptoms with lumbar root compression are well known in the classic but rather rare cauda equina syndrome. However, micturition difficulties seem to be far more frequent in lumbar root compression syndromes. METHODS: One hundred eight male patients admitted for surgery for lumbar disc herniation or spinal stenosis were investigated with an extensive questionnaire about their micturition. RESULTS: Fifty-five percent had significant lower urinary tract symptoms. Eighty percent of the patients with spinal stenosis had symptoms. Thirty-three patients had irritative symptoms, 36 had obstructive symptoms, and 23 had retention symptoms. Twenty-four had severe symptoms. Median compression resulted in more symptoms than paramedian compression. There was no correlation between age, level of compression, drug intake, or pain score and lower urinary tract symptoms. CONCLUSIONS: Lower urinary tract symptoms of mixed type occur with a high prevalence in male patients with lumbar root compression syndromes referred for neurosurgical evaluation and treatment. PMID- 9399458 TI - Pediatric chance fracture associated with pedicle screw use: a case report. AB - STUDY DESIGN: A previously undescribed clinic entity is presented, along with suggestions to prevent its reoccurrence. OBJECTIVE: To identify a potential pitfall in the use of pedicle screw instrumentation in trauma cases. Also, to emphasize the need to identify and treat noncontiguous spinal fractures. SUMMARY OF BACKGROUND DATA: No previous cases have yet been described with this particular complication, which would be remedied easily with established methods. Pedicle screw instrumentation previously has been associated primarily with complications due to aberrant screw insertion and injury to adjoining tissues, or due to fracture of the construct itself in the absence of fusion formation. METHODS: A 15-year-old girl suffered a traumatic T12-L1 fracture dislocation and paraplegia. After pedicle screw instrumentation, her apparently benign L3 fracture progressed to a severely displaced Chance fracture. This was repaired with caudal laminar hook compression instrumentation. RESULTS: Postoperatively, at a 1-year follow-up, the patient's spinal deformity was completely alleviated, though she remains paraplegic. CONCLUSIONS: Unstable traumatic spinal injuries treated with pedicular instrumentation should have additional laminar hook compression configuration reinforcement at the ends of the constructs to prevent further stress-induced injury from the screws alone. Instrumentation constructs should not end at even minimally fractured levels. PMID- 9399459 TI - Current status of percutaneous mechanical thrombectomy. Part I. General principles. PMID- 9399461 TI - Distal embolization during thrombectomy with use of the hydrolyser (hydrodynamic thrombectomy catheter): in vitro testing. AB - PURPOSE: To evaluate distal embolization while using the Hydrolyser (hydrodynamic thrombectomy catheter) with special attention to the severity of the stenosis and temporary distal or proximal flow obstruction. MATERIALS AND METHODS: The Hydrolyser procedure was assessed in plastic tubes (5-8 mm) with a 70% or 90% diameter stenosis with or without temporary distal flow obstruction and a 72-hour old clot proximal to the stenosis. The weight of the embolized particles was established after passage through filters of 1,000, 500, 100, and 10 microm. To evaluate the influence of the absolute inner diameter of the stenosis 1.0-, 2.1-, and 3.0-mm stenoses were compared in 10-mm tubes. RESULTS: Thrombus removal was greater than 99.9% in all but one of the cases in the 5-8-mm tubes. Embolization with a weight of more than 1 mg was only found in tubes with a relative stenosis of 70% and a stenosis inner diameter of greater than 1.5 mm. There was a positive relationship between inner diameter of the stenosis and the amount of distal embolization. In the presence of a proximal or distal temporary flow obstruction during thrombectomy, no distal embolization greater than 1 mg was found. CONCLUSION: In this in vitro study, the Hydrolyser thrombectomy device demonstrated minimal distal embolization. The amount of distal embolization that did occur was related to the absolute stenosis diameter and could be prevented by a severe distal stenosis and/or a temporary proximal or distal flow obstruction. PMID- 9399460 TI - Mechanical thrombectomy in acute and subacute thrombosis with use of the Amplatz device: arterial and venous applications. AB - PURPOSE: To perform a feasibility study of the Amplatz Thrombectomy Device (ATD) in a variety of vascular territories with acute or subacute thrombosis. MATERIALS AND METHODS: Thirteen patients (mean age, 44.6 years) with multiple risk factors who had acute/subacute thrombosis of the inferior vena cava (IVC) and iliac veins (n = 3), superior vena cava (SVC) and/or subclavian veins (n = 3), lower extremity polytetrafluoroethylene (PTFE) graft (n = 2), iliac artery (n = 2), portal vein and transjugular intrahepatic portosystemic shunt (TIPS) (n = 2), and an IVC to pulmonary artery Fontan conduit (n = 1), were treated by means of mechanical thrombectomy with use of the ATD. Thrombolysis failed to recanalize the vessels when used before thrombectomy for 12-34 hours in three patients, and was contraindicated in three other patients. Thrombolysis was used as a complement to the ATD procedure in five patients. RESULTS: Technical success was achieved in 11 patients, and procedure success was achieved in 10 patients. Failure was observed in the remaining three patients. One patient with a PTFE graft was successfully declotted but thrombosis occurred 2 weeks later, requiring surgery. The other patient with a PTFE graft did not improve and needed surgery to declot and treat the distal anastomosis and distal circulation. The two patients with an occluded iliac artery underwent successful declotting but rethrombosis occurred in one shortly after the procedure requiring thrombolytic therapy. One patient with TIPS thrombosis improved and another patient with a thrombosed portal vein did not improve after thrombectomy. CONCLUSION: The ATD is useful for recanalization of acute/subacute clotted native vessels and grafts. The application of the device is broad, and although declotting can be achieved in most cases, long-term success may be limited by anatomical and technical problems of the grafts and multifactorial clinical problems of severely sick patients, as was the case in the series. The use of additional thrombolytic therapy may be necessary in a number of patients. PMID- 9399462 TI - Rheolytic thrombectomy with use of the AngioJet-F105 catheter: preclinical evaluation of safety. AB - PURPOSE: A preclinical evaluation of the safety of the AngioJet-F105 rheolytic thrombectomy catheter. MATERIALS AND METHODS: The AngioJet-F105 catheter uses multiple retrograde high-speed fluid jets impinging on a primary aspiration lumen to create a hydrodynamic recirculation vortex that traps and fragments adjacent thrombus, with simultaneous evacuation of the resulting debris through the aspiration lumen. The effect of the AngioJet on treated vessels was evaluated in 10 canines. Vascular integrity on histopathologic examination and endothelial coverage on scanning electron microscopic study were examined in 15 vessel segments treated with the AngioJet-F105 catheter, compared with four vessel segments subjected to the Fogarty balloon maneuver, and 10 untreated vessel segments. The size distribution of particulate debris, upstream and downstream, after thrombectomy was determined in a flow-circuit model simulating the superficial femoral artery. Aliquots from the downstream effluent were then injected into the renal arteries of two healthy canines. RESULTS: The device caused only minimal focal endothelial denudation and no significant deep injury. No significant difference in endothelial coverage occurred in AngioJet-treated vessel segments compared to untreated control vessels (mean +/- standard deviation: 88.0% +/- 7.9% vs 89.7% +/- 11.6%, P = .77). Vessels treated with the Fogarty balloon pullback maneuver had significantly less residual endothelial coverage (58.0% +/- 8.0%, P < .03). Particulate microemboli in the effluent of the flow model accounted for 12% of the initial thrombus volume (0% > 100 microm, 99.83% < or = 10 microm). Histopathologic evaluation of the four renal beds injected with the resulting debris demonstrated no signs of necrosis. A moderate transient increase in plasma-free hemoglobin occurred, with a mild corresponding decrease in hematocrit. CONCLUSIONS: The AngioJet-F105 catheter resulted in only mild and focal injury to the treated vessels. The vast majority of resulting particulate debris consist of microscopic particles, without significant ischemic effect. PMID- 9399463 TI - Acute and delayed outcomes of mechanical thrombectomy with use of the steerable Amplatz thrombectomy device in a model of subacute inferior vena cava thrombosis. AB - PURPOSE: To study the efficacy and delayed outcome of mechanical thrombectomy with the Amplatz thrombectomy device (ATD) in an experimental model of subacute inferior vena cava (IVC) thrombosis. MATERIALS AND METHODS: Mechanical thrombectomy was performed in 23 dogs with subacute infrarenal IVC thrombosis (6 15 days old). Heparin was administered during thrombectomy in all procedures (activated clotting time > or = 300 sec). Thirteen animals were killed immediately after thrombectomy, and the remaining 10 were allowed to survive for up to 1 month with no anticoagulation therapy. RESULTS: Venographic patency of the IVC was restored in all animals, although residual mural thrombus remained in nine dogs (< 20% narrowing in seven, 20%-30% narrowing in two). No histopathologic evidence of mechanical wall disruption attributed to mechanical thrombectomy was seen. However, foci of organizing residual thrombus with associated transmural phlebitic changes with round-cellular infiltration were present in all acute specimens, including those appearing clear at venography. Venography at 1 week or 1 month after thrombectomy showed IVC rethrombosis in eight dogs (80%) who were not receiving anticoagulants. During mechanical thrombectomy, a small increase in mean pulmonary artery pressure occurred, with a corresponding decrease in systemic arterial oxygen saturation. No acute emboli were noted on the post-thrombectomy pulmonary angiograms. However, histopathologic examination of acutely explanted lungs in 11 animals showed arteriolar microemboli (100-500 microm) in four. CONCLUSION: Mechanical thrombectomy with use of the ATD can effectively clear subacute IVC thrombus. However, rethrombosis is common and may be due to the high prevalence of phlebitis and residual thrombus. Anticoagulation may need to be continued after successful thrombectomy to prevent progression of residual thrombus and allow mural phlebitic changes to subside. PMID- 9399464 TI - Guide wire directed manipulation of malfunctioning peritoneal dialysis catheters: a critical analysis. AB - PURPOSE: To evaluate patency rates after guide wire directed manipulation of malfunctioning continuous ambulatory peritoneal dialysis (CAPD) catheters. MATERIALS AND METHODS: During a 58-month period, 23 patients underwent 34 outpatient guide wire directed manipulations of their CAPD catheter to improve function (n = 30) or reduce pain and improve function (n = 4) during dialysis. Catheter patency rates were subsequently determined by review of departmental, hospital, and dialysis center charts; procedural reports; and patient telephone interviews. RESULTS: Among 12 patients who underwent a single guide wire directed manipulation, long-term (> 30 days) catheter patency was achieved in seven (58%). With use of the Kaplan-Meier survival method, the 3-, 6-, and 12-month probability of patency after a single guide wire manipulation was 0.61, 0.54, and 0.11, respectively. The mean duration of patency achieved in this group was 131 days (range, 2-421 days). In those patients (n = 8) who underwent multiple catheter manipulations (n = 19), 11 (58%) procedures resulted in long-term patency, with each patient (100%) achieving at least one such period. The Kaplan Meier survival method determined the probability of patency in this group at 3, 6, and 12 months to be 0.75, 0.69, and 0.54, respectively. The mean secondary catheter patency was 235 days (range, 2-646 days). Overall, 75% of patients followed up achieved at least one period of long-term catheter patency during the time of this study. One (3%) episode of postprocedure peritonitis occurred. CONCLUSION: Guide wire directed CAPD catheter manipulation is a relatively simple outpatient procedure that restores long-term catheter function for most patients with minimal risk for a major complication. Patients with nonfunctioning CAPD catheters who do not have peritonitis or sepsis will most likely benefit from at least one attempt at radiologic manipulation of their catheter. PMID- 9399465 TI - Prospective randomized trial of a metallic intravascular stent in hemodialysis graft maintenance. AB - PURPOSE: To evaluate percutaneous transluminal angioplasty (PTA) alone versus PTA and flexible self-expanding stent placement for the management of hemodialysis access graft stenoses. MATERIALS AND METHODS: Thirty-seven grafts in 34 patients were evaluated for abnormal intradialytic parameters (n = 27) or occlusion (n = 10). Angiography identified stenoses (mean, 69%; range, 50%-95%) at or within 3 cm of the vein-graft junction (70%) or in the peripheral outflow vein (30%) that had recurred within a 6-month period after previous PTA. They were randomized to PTA alone (n = 20) or PTA with Wallstent (n = 17). Additional lesions were treated by PTA alone, and a mean of 1.4 (range, 1-3) lesions were treated per patient. Significant differences existed in the mean number of previous accesses (1.8 and 0.8 in the PTA and stent groups, respectively) and in the mean number of previous interventions in the current access (1.8 and 2.9, respectively). End points were subsequent radiologic or surgical intervention, transplantation, and death. RESULTS: Technical success was 100% (mean residual stenosis, 12%; range, 0%-30%). The primary patency of 128 days and secondary patency of 431 days were similar for both groups. Secondary patency required a mean of 1.8 and 1.6 additional interventions for the PTA and stent groups, respectively. The adjunctive stent placement increased the cost of the procedure by 90%. CONCLUSION: Despite significant added costs, there was no advantage to stent placement for recurrent peripheral hemodialysis graft stenoses that were already adequately dilated with balloon angioplasty. PMID- 9399466 TI - Wallstents and Craggstents in hemodialysis grafts and fistulas: results for selective indications. AB - PURPOSE: To report the value of selective placement of self-expandable stents (Wallstent and Craggstent) for the treatment of limitations and, occasionally, of complications of dilation in hemodialysis access, and especially for delaying restenosis. MATERIALS AND METHODS: This is a retrospective study of a 7-year period, during which 41 Wallstents and 11 Craggstents were placed in 26 polytetrafluoroethylene (PTFE) grafts, 15 native fistulas, and nine central veins of 47 patients. The indications were stenosis recoil (n = 13), recurrent restenosis within 6 months (n = 33), restenosis after 6 months (n = 3), and acute angioplasty-induced rupture (n = 1). Restenosis after stent placement necessitated redilation and percutaneous declotting and 10 additional stent placements. RESULTS: Two initial misplacements were corrected immediately. Primary patency rates for PTFE grafts were 58% +/- 10% at 6 months and 23% +/- 10% at 1 year, respectively. Secondary patency rates were 100% at 6 months and 88% +/- 8% at 1 year, respectively. For native fistulas, primary patency rates were 47% +/- 12% at 6 months and 20% +/- 18% at 1 year. Secondary patency rates were 95% +/- 6% at 6 months and 79% +/- 14% at 1 year. It was necessary to reintervene after stent placement to maintain or to restore patency every 9 months for PTFE grafts and every 7.3 months for native fistulas. When stents were placed for treatment of early recurring restenosis, the mean interval between radiologic interventions (redilations or declottings) performed to maintain or to restore patency before stent placement was multiplied by 2.1 after stent placement for both grafts (3.2 months increased to 6.9, P < .01) and native fistulas (2.9 months increased to 6.2, P < .02). CONCLUSIONS: Wallstents and Craggstents are valuable for the treatment of failure of regular dilation and they double the intervals between reinterventions for early (< 6 months) recurring stenoses in PTFE grafts and native fistulas. PMID- 9399467 TI - Experience with 100 consecutive central venous access arm ports placed by interventional radiologists. AB - PURPOSE: This study reports the authors' experience with long-term follow-up of 100 consecutive peripherally inserted, subcutaneous arm ports for central venous access. MATERIALS AND METHODS: One hundred patients with subcutaneous arm ports inserted by interventional radiologists were retrospectively studied. Data were collected from the patients' medical records and from telephone canvassing. Using each insertion period as an observation, the complication rates per 100 catheter days were determined with 95% confidence intervals (CIs). RESULTS: One hundred subcutaneously implanted ports were placed in 98 patients; three devices (three patients) were lost to follow-up, leaving 97 devices in 95 patients. Total exposure time was 23,842 days (mean, 246 days; range, 2-865 days). Seven infectious and two noninfectious complications occurred with seven (7.2%) devices in six patients (6.3%), yielding 0.038 complications per 100 catheter days at risk (95% CI; 0.011-0.069) and 0.029 infections per 100 catheter days at risk (95% CI; 0.008-0.058). A successful clinical outcome was defined as a functional port at removal, time of death, or at study closure (minimum of 6 months of follow-up), which was not removed because of a complication. This successful outcome was achieved in 91 ports (93.8%). Procedural-related complications, defined as those occurring up to 30 days after insertion, occurred in only one port (thrombophlebitis and catheter tip infection-day 9). All other patients received several months of service from their port. Fifteen devices were placed in 13 patients with HIV for 3,486 days, with a total complication rate of 0.11 per 100 catheter days (95% CI; 0.0-0.28), all of which were infections. Devices in HIV-positive patients were associated with higher total complication (20% vs 4.9%) and infection rates (20% vs 3.7%) than devices in patients without HIV infection. This gives a relative risk 8.17 x (P = .04) greater for infectious complications for devices placed in HIV-infected individuals. CONCLUSIONS: Subcutaneous arm ports placed by interventional radiologists are effective for central venous access with excellent functionality (93.8% achieved a successful long-term outcome) and a very low procedural complication rate. Although infections were more frequent in HIV-infected individuals, these devices are associated with a very low incidence of both immediate and long-term complications, including infection, for all patients. PMID- 9399468 TI - Outcome of 350 implanted chest ports placed by interventional radiologists. AB - PURPOSE: To determine the outcome of implanted chest ports placed by interventional radiologists. MATERIALS AND METHODS: Between June 1993 and July 1996, a single institution placed 350 implanted chest ports in 346 patients by means of the subclavian vein approach. The medical records of these patients were reviewed to determine the outcome of the ports. Ports were implanted for chemotherapy (n = 341), blood transfusion (n = 7), or antibiotics (n = 2). RESULTS: Immediate complications were seven (2%) pneumothoraces and one (0.3%) hematoma. Four (1.1%) of the pneumothoraces necessitated hospital admission and treatment with a chest tube. The remaining three were managed on an outpatient basis. One was successfully treated in the interventional suite by catheter suction. Two pneumothoraces were observed and resolved spontaneously. Mean time of patient follow-up was 260 days (range, 22-929 days). Total time of follow-up was 91,000 catheter days. Delayed complications were 10 cases of thrombosis (2.9% or 0.11 per 1,000 catheter days) of the subclavian vein, four infections (1.1% or 0.04 per 1,000 catheter days), four catheter coiling or tip malpositions (1.1% or 0.04 per 1,000 catheter days), three catheter occlusions (0.9% or 0.03 per 1,000 catheter days), and one catheter leak (0.3% or 0.01 per 1,000 catheter days). Six (1.7%) ports had to be removed as a result of a delayed complication. CONCLUSION: Chest port implantation by interventional radiologists within the radiology department is a successful and safe procedure with complication rates equivalent to, or lower than, those reported in surgical placement series. PMID- 9399469 TI - Mural delivery of iloprost with use of hydrogel-coated balloon catheters suppresses local platelet aggregation. AB - PURPOSE: To develop reproducible and quantifiable methods for mural delivery of iloprost, a potent agent against platelet aggregation, with use of hydrogel coated angioplasty balloons, and to determine the in vivo effect of direct iloprost delivery on platelet aggregation at the angioplasty site. MATERIALS AND METHODS: Drug loading of tritiated iloprost from an immersion solution onto hydrogel-coated balloons was evaluated as a function of balloon size (3 mm x 2 cm, 6 mm x 2 cm, 8 mm x 3 cm; n = 4 each), drug concentration (0.0715 mg/mL, 0.1072 mg/mL, 0.1430 mg/mL; n = 3 each), and duration of immersion (40 seconds, 60 seconds, 120 seconds; n = 3 each). In another set of experiments, optimal drying methods were tested to minimize drug loss within a protective delivery sheath (n = 3 each). Ex vivo angioplasty was performed on excised swine arteries to estimate how much of the drug present on the balloon could be delivered to the wall (n = 3 iliac segments). Finally, in vivo angioplasty was performed in three Yorkshire pigs (n = 6 iloprost-treated and 6 control arteries) and indium-111 labeled platelet aggregation was measured at these sites, which were harvested 1 hour after the procedure. RESULTS: In the initial set of experiments, the authors found that the volume of drug loaded is determined by the wet-volume of the hydrogel coating, that the majority of volume loading occurs within the first 2 minutes, and that the volume uptake is independent of the drug concentration. The optimal drying method resulting in the least loss of iloprost within the sheath (only 4%) was prolonged drying (5 hours) under ambient conditions. Ex vivo angioplasty experiments showed that approximately 33% of the drug present on the balloon can be delivered to the wall. Finally, in vivo experiments showed that platelet aggregation is significantly suppressed at treated sites (by approximately 33% compared to control sites; P = .03) by minuscule mural doses of iloprost (roughly estimated at under 1 microg). CONCLUSION: Quantifiable and reproducible methods for loading iloprost onto hydrogel-coated angioplasty balloons were developed. The best of these methods was able to deliver enough iloprost into the wall to significantly reduce local platelet aggregation. PMID- 9399470 TI - Transcatheter venous thrombolysis--pitfalls and pratfalls: a case discussion of indications, technique, and alternatives. PMID- 9399471 TI - Detection of implanted metallic devices by airport security. AB - PURPOSE: Permanent metallic implants are commonly placed by interventional radiologists. The authors evaluate the ability of these and various orthopedic devices to be detected by airport security metal detectors. MATERIALS AND METHODS: A variety of commonly placed radiologic and orthopedic implants were evaluated by three types of metal detectors at Toronto International Airport. RESULTS: No radiologic devices were detected by walk-through detectors. Metal containing ports were the only device to set off the hand-held scanners. CONCLUSION: Radiology patients can be reassured that their devices will not set off walk-through airport security metal detectors, however, some metallic ports may be detected by hand-held scanning devices. PMID- 9399472 TI - Digital subtraction versus film-screen angiography for detecting acute pulmonary emboli: evaluation in a porcine model. AB - PURPOSE: To compare the diagnostic performance of digital subtraction angiography (DSA) to that of film-screen angiography (FSA) for detecting acute pulmonary embolism (PE) in a porcine model. MATERIALS AND METHODS: DSA and FSA were performed in 13 pigs before and after central venous administration of autologous emboli. Results were compared to findings at necropsy with use of ex vivo pulmonary angiography to guide pathologic sectioning. The sensitivity and predictive value of a positive case for detecting each embolus were computed for each pulmonary artery branch order and compared with use of 95% confidence intervals. Interobserver variability among three readers for individual PE detection was calculated. RESULTS: Pathologic examination of the lungs revealed 100 total PEs (location by vessel order: 1st = 1, 2nd = 0, 3rd = 15, 4th = 32, > 5th = 52). On average, FSA review identified 72 (72%) emboli and DSA review, 65 (65%). There was no significant difference in sensitivity or predictive value of a positive case between DSA and FSA for detecting emboli (P > .05). There was similar agreement among readers for individual PE detection with DSA (mean, 84%) and FSA (mean, 80%). CONCLUSION: The diagnostic performance of DSA is equivalent to that of FSA for detecting emboli in porcine PA branches. Interobserver agreement for individual PE detection is similar for both imaging techniques. PMID- 9399473 TI - Life-threatening gastrointestinal hemorrhage secondary to nonmesenteric sources. PMID- 9399474 TI - IVC filter tilt and asymmetry: comparison of the over-the-wire stainless-steel and titanium Greenfield IVC filters. AB - PURPOSE: A comparison of tilting, caval coverage, asymmetry, and insertion problems with the over-the-wire stainless-steel and titanium versions of the Greenfield filter. MATERIALS AND METHODS: The study compared 104 stainless-steel and 141 titanium Greenfield inferior vena cava (IVC) filter insertions. The angle the sheath and deployed filter made relative to the cava, as well as filter strut distribution, were determined from spot films. The proportionate caval coverage was computed from the cavogram (anteroposterior projection). Mean filter tilts, subgrouped by insertion site, and caval coverage were compared with the Student t test, whereas strut patterns were analyzed with a contingency table. RESULTS: The filter caval and sheath caval angles correlated. The filter caval angles varied with insertion site, but were lowest with a right jugular approach. Caval coverage was identical with both designs. The stainless-steel version resulted in a more uniform distribution of struts in comparison with the titanium version. The incidence of insertion problems was not significantly different between the filter types. CONCLUSIONS: While IVC filter tilting was not improved with the newer design, the pattern of struts was more uniformly symmetric with the stainless-steel device. The right jugular insertion site was associated with the lowest filter caval angles and the most symmetric pattern of struts. PMID- 9399475 TI - Distal embolization from an unsuspected external iliac artery pseudoaneurysm: diagnosis during urokinase infusion. PMID- 9399476 TI - Treatment of hemoptysis in Behcet syndrome with pulmonary and bronchial embolization. PMID- 9399477 TI - Treatment of chronic iliac artery occlusions with guide wire recanalization and primary stent placement. AB - PURPOSE: To evaluate the results of primary stent placement without initial thrombolysis in the treatment of iliac occlusions. MATERIALS AND METHODS: During a 3-year period, 61 iliac artery occlusions were treated in 59 patients. The mean length of the occluded segment was 10 cm (range, 4-25 cm). The occluded arteries were treated with primary placement of self-expandable metallic stents. RESULTS: Successful recanalization with primary stent placement was possible in 56 of 61 occlusions (92% technical success rate). Mean Doppler ankle/brachial index increased from 0.51 to 0.90 immediately after treatment and was 0.91 on the last follow-up (P < .05). Primary patency rate at 24 months was 73%, and secondary patency rate was 88%. Procedural complications included distal embolization (n = 4) and an episode of massive intra-abdominal bleeding. Three patients developed a hematoma at the puncture site that did not require additional therapy. Late complications included stent occlusion (n = 9) and significant stenosis related to intimal hyperplasia (n = 1). Mean follow-up period was 29 months (range, 7-55 months). CONCLUSION: Primary stent placement is an effective therapeutic option for iliac artery occlusions. PMID- 9399478 TI - Endovascular stents covered with pre-expanded polytetrafluoroethylene for treatment of iliac artery aneurysms and fistulas. AB - PURPOSE: This report describes the early clinical experience with use of a transluminally placed endovascular graft (TPEG) covered with pre-expanded polytetrafluoroethylene (PTFE) to treat iliac artery aneurysms and fistulas. MATERIALS AND METHODS: Eight patients with iliac artery aneurysms (n = 7) and common iliac artery to common iliac vein fistula (n = 1) were treated with TPEGs. The iliac artery aneurysms were either common iliac (n = 6) or hypogastric (n = 1). All of the patients had significant comorbid diseases. The TPEG devices were made with pre-expanded PTFE sutured to Palmaz stents and delivered through 10- or 12-F sheaths. RESULTS: The aneurysms were successfully excluded in six of seven patients and the one iliac artery-to-vein fistula was successfully occluded. There were no immediate procedural complications related to the TPEG devices. Follow-up was limited (mean, 12 months), but no stenoses or occlusions of the TPEG devices were detected. The one failure was probably due to the marked tortuousity of the iliac artery, which prevented an adequate seal. CONCLUSION: In the authors' early clinical experience, the use of TPEG devices with pre-expanded PTFE successfully treated iliac artery aneurysms and an iliac artery-to-vein fistula. Although the results are encouraging, longer follow-up is necessary to better evaluate this type of treatment. PMID- 9399479 TI - Treatment of traumatic carotid pseudoaneurysm with endovascular stent placement. PMID- 9399480 TI - Use of a Palmaz stent deployed in the superior mesenteric artery for chronic mesenteric ischemia. PMID- 9399481 TI - Use of CT-guided marking of the portal vein in creation of 150 transjugular intrahepatic portosystemic shunts. PMID- 9399482 TI - Percutaneous treatment of an iliac artery pseudoaneurysm associated with a stent graft. PMID- 9399483 TI - Vascular or interventional procedures in patients with diabetes. PMID- 9399484 TI - Dislodgment of a stainless-steel Greenfield filter during exchange of a central venous catheter. PMID- 9399485 TI - Venous injection to assess for thoracic aortic trauma. PMID- 9399486 TI - IS6110 fingerprinting of drug-resistant Mycobacterium tuberculosis strains isolated in Germany during 1995. AB - The epidemiological relatedness of drug-resistant Mycobacterium tuberculosis strains isolated in Germany in 1995 was evaluated by the standardized IS6110 fingerprinting method. Altogether, 196 M. tuberculosis isolates from 167 patients were analyzed. A large degree of IS6110 polymorphism was found, ranging from 1 to 20 copies. Multiple isolates from one patient generally remained stable over a period of up to 1 year. However, one strain showed an additional fragment 7 months after the first isolate was obtained. Isolates from 55 patients (33%) showed identical fingerprint patterns or fingerprint patterns that differed only in one band, and thus they were clustered in 22 fingerprint groups. Specific transmission links could be established between members of four groups, e.g., transmission by family contacts. In one case, transmission of a multidrug resistant strain to a patient initially infected with a drug-susceptible strain could be shown. Besides these fingerprint groups, 30 of the 167 isolates (approximately 18%) could be grouped in two fingerprint clusters with a similarity of at least 78%. Approximately 60% of the patients of these two clusters were known to be immigrants from the former Soviet Union, and one patient is still living in Belarus. In conclusion, our results indicate that (i) transmission of drug-resistant strains contributes substantially to the emergence of drug-resistant tuberculosis in Germany and (ii) drug-resistant M. tuberculosis strains were presumably carried over from the former Soviet Union to Germany by immigrants. PMID- 9399487 TI - Differentiation of Helicobacter pylori strains directly from gastric biopsy specimens by PCR-based restriction fragment length polymorphism analysis without culture. AB - Recent studies have shown the usefulness of PCR-based restriction fragment length polymorphism (RFLP) analysis for differentiating Helicobacter pylori strains isolated by culture. For this study, a PCR-based RFLP assay was developed for directly typing H. pylori strains from gastric biopsy specimens. Nineteen gastric biopsy specimens obtained from patients undergoing endoscopy for gastrointestinal complaints were cultured for isolation of H. pylori. Genomic DNA preparations from these gastric biopsy specimens and the corresponding H. pylori isolates were tested by our PCR-based RFLP assay. The 1,179-bp H. pylori DNA fragments amplified by the PCR assay were digested with the restriction enzymes HhaI, MboI, and AluI and analyzed by agarose gel electrophoresis. HhaI, MboI, and AluI digestion produced 11, 10, and 6 distinguishable digestion patterns, respectively, from the 19 H. pylori isolates tested and generated 13, 11, and 6 different patterns, respectively, from the 19 gastric biopsy specimens. The patterns from 13 of the 19 gastric biopsy specimens matched those of the H. pylori isolates from the corresponding patients. The patterns from the remaining six biopsy specimens appeared to represent infection by two strains of H. pylori; the pattern of one strain was identical to that of the isolate from the corresponding patient. By combining all the restriction enzyme digestion patterns obtained by using HhaI, MboI, and AluI, we observed 19 distinct RFLP patterns from the 19 specimens. The results suggest that the PCR-based RFLP analysis method may be useful as a primary technique to identify and distinguish H. pylori strains directly from gastric biopsy specimens without culture of the organisms. PMID- 9399488 TI - Fecal carriage of vancomycin-resistant enterococci in hospitalized patients and those living in the community in The Netherlands. AB - In order to determine the prevalence of vancomycin-resistant enterococci (VRE) in The Netherlands, 624 hospitalized patients from intensive care units or hemato oncology wards in nine hospitals and 200 patients living in the community were screened for VRE colonization. Enterococci were found in 49% of the hospitalized patients and in 80% of the patients living in the community. Of these strains, 43 and 32%, respectively, were Enterococcus faecium. VRE were isolated from 12 of 624 (2%) and 4 of 200 (2%) hospitalized patients and patients living in the community, respectively. PCR analysis of these 16 strains and 11 additional clinical VRE isolates from one of the participating hospitals revealed 24 vanA gene-containing, 1 vanB gene-containing, and 2 vanC1 gene-containing strains. All strains were cross-resistant to avoparcin but were sensitive to the novel glycopeptide antibiotic LY333328. Genotyping of the strains by arbitrarily primed PCR and pulsed-field gel electrophoresis revealed a high degree of genetic heterogeneity. This underscores a lack of hospital-driven endemicity of VRE clones. It is suggested that the VRE in hospitalized patients have originated from unknown sources in the community. PMID- 9399489 TI - Molecular tracking of Candida albicans in a neonatal intensive care unit: long term colonizations versus catheter-related infections. AB - Nosocomial neonatal candidiasis is a major problem in infants requiring intensive therapy. The subjects of this retrospective study were nine preterm infants admitted to the neonatal intensive care unit of the Hospital Central de Asturias between March 1993 and August 1994. The infants were infected with or colonized by Candida albicans. Five patients developed C. albicans bloodstream infections. A total of 36 isolates (including isolates from catheters and parenteral nutrition) were examined for molecular relatedness by PCR fingerprinting and restriction fragment length polymorphism (RFLP) analysis. The core sequence of phage M13 was used as a single primer in the PCR-based fingerprinting procedure, and RFLP analysis was performed with C. albicans-specific DNA probe 27A. Both techniques were evaluated with a panel of eight C. albicans reference strains, and each technique showed eight different patterns. With the 36 isolates from neonates, each technique enabled us to identify by PCR and RFLP analysis seven and six different patterns, respectively. The combination of these two methods (composite DNA type) identified eight different profiles. A strain with one of these profiles was present in three patients and in their respective catheters. Patients infected with or colonized by this isolate profile were clustered in time. Among the other patients, each patient was infected over time and at multiple anatomic sites with a C. albicans strain with a distinct DNA type. We conclude that C. albicans was most commonly producing long-term colonizations, although horizontal transmission probably due to catheters also occurred. PMID- 9399490 TI - Racial tropism of a highly toxic clone of Actinobacillus actinomycetemcomitans associated with juvenile periodontitis. AB - Actinobacillus actinomycetemcomitans strains with enhanced levels of production of leukotoxin are characterized by a 530-bp deletion from the promoter region of the leukotoxin gene operon. Previous isolates with this deletion constituted a single clone belonging to serotype b, although they displayed minor differences among each other. We have analyzed the geographic dissemination of this clone by examining 326 A. actinomycetemcomitans isolates from healthy and periodontally diseased individuals as well as from patients with different types of extraoral infections originating from countries worldwide. A total of 38 isolates, all belonging to the same clone, showed the 530-bp deletion. Comparison of a 440-bp sequence from the promoter region of the leukotoxin gene operon from 10 of these strains revealed complete identity, which indicates that the deletion originates from a single mutational event. This particular clone was exclusively associated with localized juvenile periodontitis (LJP). In at least 12 of 28 families from which the clone was isolated, more than one family member had LJP. Notably, all the subjects carrying this clone had a genetic affiliation with the African population. These observations suggest that juvenile periodontitis in some adolescents with an African origin is associated with a disseminating clone of A. actinomycetemcomitans. PMID- 9399491 TI - Immunoblot analysis of Leishmania panamensis antigens in sera of patients with American cutaneous leishmaniasis. AB - Sera from 86 Colombian patients with parasitologically confirmed cutaneous leishmaniasis were studied by immunoblot analysis in order to identify a specific pattern for Leishmania infection. A soluble extract of Leishmania panamensis was used as the antigen. Sera from patients with Chagas' disease and sera from patients with no record of infection with trypanosomatids were also studied. The sera from patients with cutaneous leishmaniasis specifically recognized fractions of 120 kDa (76.7%), 123 and 129 kDa (69.7%), 138 kDa (61.6%), 141 kDa (53.4%), and 78 kDa (44.1%). No band common to all patients infected with Leishmania parasites was found at the time of diagnosis. Likewise, the pattern of immunoblot change after the patients were treated and apparently cured with meglumine antimoniate (Glucantime) was studied by evaluating sera pretreatment and 1 year posttreatment. Only minor changes in the color intensity at the same serum dilution between pre- and posttreatment sera were found, although the antibody titers by indirect immunofluorescence were negative for the posttreatment sample. This study shows that Western blot analysis is a more sensitive test for the detection of anti-Leishmania antibodies. However, the importance of whether the presence of antibodies correlates with the presence of Leishmania antigens could not be resolved by the data obtained from this study. PMID- 9399492 TI - Distinctive IS200 insertion between gyrA and rcsC genes in Salmonella typhi. AB - While probing the vicinity of ompC, a copy of the IS200 insertion element was found between the gyrA and rcsC genes of Salmonella typhi, the causal agent of typhoid fever. This distinctive feature was conserved throughout 63 S. typhi isolates of different geographical origins and was absent from 46 other Salmonella serotypes, including those most associated with human infections, as well as from 19 other enteric bacteria. Furthermore, the location of this IS200 copy corresponds to a constant band, present throughout the 14 PstI S. typhi IS200 fingerprints, encompassing several Vi phage types. Interestingly, an apparently unrelated serotype not frequently associated with human disease, Salmonella weltevreden, contained an IS200 copy at the same position, although it was accompanied by an additional segment of cryptic DNA. On the basis of these findings, PCR assays were designed for molecular typing of S. typhi, and these are potentially useful in studying the epidemiology of typhoid fever. PMID- 9399493 TI - Identification of Legionella species by lipopolysaccharide antigen pattern. AB - Electrophoretic analysis of lipopolysaccharide (LPS) extracts from 430 previously serotyped Legionella isolates and 28 American Type Culture Collection (ATCC) non Legionella pneumophila Legionella reference strains representing different Legionella species and serogroups has been performed. LPS was prepared from Legionella suspensions by sonication and proteinase K digestion. Following sodium dodecyl sulfate-polyacrylamide gel electrophoresis, LPS bands were either stained with silver nitrate or transferred onto a nitrocellulose membrane and detected with rabbit antibodies raised against L. pneumophila serogroup 5, which was known to cross-react with L. pneumophila serogroups 1 to 14. Silver staining revealed that each of the 28 ATCC non-L. pneumophila Legionella strains possessed an individual and characteristic LPS banding pattern. The LPS profile was defined by the molecular weight of the visualized bands and/or the individual ladder-like LPS pattern. It was demonstrated by immunoblotting that non-L. pneumophila Legionella strains did not react with the serogroup 5 antiserum, thus allowing for the differentiation between L. pneumophila and non-L. pneumophila species. PMID- 9399494 TI - Use of serum-specific immunoglobulins A and G for detection of Helicobacter pylori infection in patients with chronic gastritis by immunoblot analysis. AB - Multiple invasive and noninvasive tests for detecting Helicobacter pylori infection are available. The current "gold standard" for the diagnosis of H. pylori infection requires histology and the rapid urease test. Our aim was to test the performance of immunoglobulin A (IgA) and IgG immunoblot assays in comparison with that of the gold standard tests for the diagnosis of H. pylori infection. Ninety patients who underwent gastroscopy were analyzed in a prospective study. Fifty-nine of them were defined to be H. pylori positive by the gold standard tests. The IgA and IgG immunoblot assays correctly identified H. pylori infection in 17 and 58 of these patients, respectively, indicating that determination of IgA antibodies seems to be of low diagnostic value for H. pylori infection. In contrast, the sensitivity and specificity of the IgG immunoblot assay were 98 and 71%, respectively, with positive and negative predictive values of 87 and 96%, respectively. Therefore, the IgG immunoblot assay proved to be a sensitive and useful, noninvasive test for the diagnosis of H. pylori infection. PMID- 9399495 TI - Antigenic variation of core, NS3, and NS5 proteins among genotypes of hepatitis C virus. AB - Assays that detect antibody to hepatitis C virus (HCV) are used to screen blood donors and patients with hepatitis. Current enzyme-linked immunosorbent assay (ELISA)-based methods are invariably based upon antigens from expressed recombinant proteins or oligopeptides from HCV type 1. Some HCV antigens used in screening assays are coded by regions of the HCV genome that show extensive variability; therefore, HCV type 1-based assays may be less effective for the detection of antibody elicited by infection with other genotypes. In this study, we have measured antibody reactivity of sera from 110 hepatitis C patients infected with type 1b, 3a, or 4a to genotype-specific and cross-reactive epitopes present in recombinant proteins from HCV genotypes 1b (core, NS3, and NS5), 3a (NS3, NS5), and 4a (core, NS3), corresponding to those used in current third generation screening ELISAs. By comparing the serological reactivities of sera to type-homologous and type-heterologous antigens, we detected a significant type specific component to the reactivity to NS3 (61 to 77% of the total reactivity) and NS5 (60% of the total reactivity). Furthermore, despite the similarities in the amino acid sequences of the core antigens of type 1b and type 4a, we also found significantly greater reactivity to type-homologous antigens, with approximately 25% of reactivity being type specific. These findings are consistent with previous findings of fivefold weaker reactivity of sera from HCV type 2- and HCV type 3-infected blood donors in the currently used third generation ELISAs and suggest that these assays are suboptimal for screening populations in which the predominant genotype is not type 1. PMID- 9399496 TI - Multicenter study using standardized protocols and reagents for evaluation of reproducibility of PCR-based fingerprinting of Acinetobacter spp. AB - Seven laboratories in six European countries examined 40 isolates belonging to the Acinetobacter calcoaceticus-Acinetobacter baumannii complex to investigate whether standardized protocols and quality-controlled reagents could produce reliable, discriminatory, and reproducible PCR-based fingerprinting results. Four PCR protocols with different primers (primers DAF4, ERIC-2, M13, and REP1 + REP2) were used. The epidemiological conclusions reached by the participating laboratories were substantially correct, with 96.4% of the total isolate grouping allocations agreeing with the consensus view. All laboratories identified the main epidemiological clusters, and each laboratory also identified two non outbreak-related isolates. There were no significant differences between the isolate grouping results obtained by the different protocols and with the different primers. Visual comparison indicated that the standardized protocols and reagents yielded reproducible fingerprint patterns, but with some variations in particular band intensities. Minor variations in fingerprint profiles were detected, but computer-assisted analysis of PCR fingerprints obtained on agarose gels demonstrated that 88.3 to 91.6% (depending on the source of DNA) of the patterns clustered correctly, while 96.4 to 98.9% of the patterns clustered correctly following automated high-resolution laser fluorescence analysis. Correlation of the patterns for isogenic isolates ranged from 83.3 to 86.6% but was slightly better (mean correlation, 87.1%) for centrally prepared DNA extracts than for DNA extracts prepared by individual laboratories (mean correlation, 84.7%). It was concluded that independently produced PCR fingerprint patterns can be obtained reproducibly for Acinetobacter spp. at the practical level if (i) quality-controlled reagents, (ii) standardized extraction of DNA, and (iii) standardized amplification conditions are used. PMID- 9399497 TI - Influence of endocervical specimen adequacy on PCR and direct fluorescent antibody staining for detection of Chlamydia trachomatis infections. AB - The cellular quality of the endocervical swab specimen used for the detection of Chlamydia trachomatis may dramatically impact the sensitivity of the diagnostic assay used. An evaluation of the adequacy of 319 endocervical swab specimens from women attending two inner-city sexually transmitted disease and family planning clinics, as well as five high school-based family planning clinics, was performed, and the resulting data were compared with the diagnostic results obtained by both Amplicor PCR and Microtrak direct fluorescent-antibody (DFA) staining. The swab from each patient was rolled across the open circular area of a DFA slide and then used to inoculate a transport tube for PCR (Roche), after which the swab was discarded. The slides were stained and examined by epifluorescence microscopy for the presence of C. trachomatis elementary bodies and for the presence and number of cell types to determine specimen adequacy. Cellular adequacy for a cervical swab specimen was defined as the presence of one or more columnar epithelial or metaplastic epithelial cells or the presence of more than 100 erythrocytes per high-power microscopic field. Of the 319 specimens read by DFA, 204 (63.9%) were determined to be adequate. There were 34 (10.7%) positive specimens by DFA and/or PCR. Twenty-nine (9.1%) specimens were positive by PCR, 20 (6.3%) specimens were DFA positive, and 15 (4.7%) were concordantly positive by both tests. The prevalence of chlamydia among adequate specimens was 14.2% (29/204), compared to 4.3% (5/115) for inadequate specimens (P < 0.0001). Variations in specimen quality and the sensitivity of the diagnostic assay used have a significant impact on determining the prevalence of C. trachomatis in a population. PMID- 9399499 TI - Typing of Pneumocystis carinii f. sp. hominis by single-strand conformation polymorphism of four genomic regions. AB - To better investigate Pneumocystis carinii f. sp. hominis epidemiology, we have developed a molecular typing method. Because of the limited genetic variability of the P. carinii hominis genome, a multitarget approach was used. Four variable regions of the genome were amplified by PCR, polymorphism in each region was assessed by the single-strand conformation polymorphism (SSCP) technique, and the results for the four regions of each patient were combined. Bronchoalveolar lavage specimens collected from 11 patients were examined. Four patients were probably infected by a single strain, since their specimens yielded simple SSCP patterns (two bands corresponding to one allele). The combinations of these patterns were unique, suggesting that the strains which infected these patients were different. For the other seven patients, complex patterns were found (three or four bands corresponding to two alleles). The presence of more than one allele of a region in a patient is likely to be due to coinfection. Polymorphism was also assessed by sequencing, which revealed variations at nucleotide positions previously reported to vary. About half of the observed alleles had already been reported by laboratories in different countries. Multitarget typing of P. carinii hominis by PCR-SSCP should allow investigation of strain diversity and thus be useful for future epidemiological studies. PMID- 9399498 TI - Diagnostic value of the strand displacement amplification method compared to those of Roche Amplicor PCR and culture for detecting mycobacteria in sputum samples. AB - We compared the ability of the semiautomated BDProbeTec-SDA system, which uses the strand displacement amplification (SDA) method, with that of the Roche Amplicor-PCR system and the Septi-Chek AFB culture system to directly detect Mycobacterium tuberculosis complex (MTB) and other mycobacteria in sputum samples. A total of 530 sputum samples from 299 patients were examined in this study. Of the 530 samples, 129 were culture positive for acid-fast bacilli with the Septi-Chek AFB system; 95 for MTB, 29 for M. avium-M. intracellulare complex (MAC), and 5 for other mycobacteria. The BDProbeTec-SDA system detected 90 of the 95 samples culture positive for MTB (sensitivity, 94.7%), and the Amplicor-PCR system detected 85 of the 95 samples culture positive for MTB (sensitivity, 89.5%). The specificity of each system, based on the clinical diagnosis, was 99.8% for SDA and 100% for PCR, respectively. Among the 29 samples culture positive for MAC, the BDProbeTec-SDA system detected MAC in 24 samples (sensitivity, 82.8%), whereas the Amplicor-PCR system detected MAC in 23 samples (sensitivity, 79.3%). The specificities of the systems were 98.3 and 100%, respectively. The high degrees of sensitivity and specificity of the BDProbeTec SDA system suggest that it should be very useful in clinical laboratories for the rapid detection of mycobacteria in sputum samples. PMID- 9399500 TI - Analysis of an outbreak of non-phage-typeable methicillin-resistant Staphylococcus aureus by using a randomly amplified polymorphic DNA assay. AB - A cluster of methicillin-resistant Staphylococcus aureus (MRSA) infections among patients on an intensive care unit (ICU) was detected by routine infection control surveillance. In the period from 5 January to 22 June 1995, 10 patients on the ICU and a further 6 patients (5 on one ward that had received colonized patients transferred from the ICU) were affected by MRSA strains with the same antibiotic susceptibility patterns. Seven (44%) of these 16 colonized patients developed MRSA bacteremia. MRSA isolates with the same characteristics were also found on the hands of one member of the ICU staff. The isolates were untypeable by phage typing, but 15 of 17 outbreak strains analyzed genetically had identical randomly amplified polymorphic DNA (RAPD) and pulsed-field gel electrophoresis (PFGE) profiles. A single strain of MRSA that was nontypeable by phage typing and that was isolated on the ICU on 1 January and six nontypeable and epidemiologically unrelated MRSA isolates all had RAPD profiles distinct from that of the outbreak strain. Implementation of strict infection control measures stopped the further spread of MRSA on the ICU, the affected general ward, and seven other wards that received MRSA carriers from the ICU. Although nontypeable by phage typing and not previously recognized as an epidemic strain, this strain of MRSA was readily transmissible and highly virulent. RAPD typing was found to be a simple, rapid, and effective method for the epidemiological investigation of this outbreak, and performance of typing by this method was simpler and less time consuming than that of typing by PFGE. RAPD typing may have more general application for the study of S. aureus infections in hospitals. PMID- 9399501 TI - Human papillomavirus and host variables as predictors of clinical course in patients with juvenile-onset recurrent respiratory papillomatosis. AB - This study provides the first systematic evaluation of papillomavirus type and viral mutation occurring during the course of juvenile-onset recurrent respiratory papillomatosis. One hundred ninety-nine consecutive papillomas excised from 47 children between 1981 and 1996 at The New Children's Hospital in Sydney, Australia, were tested for human papillomavirus (HPV) DNA by PCR. PCR products from the viral upstream regulatory region (URR) enhancer were sequenced, and variation was related to clinical variables. Forty-four of the 47 children had HPV-induced papillomas, with type 11 accounting for 24 (55%) and type 6 accounting for 19 (43%); one (2%) was positive for either type 6 or 11. Overall, 183 (98%) of the 186 samples with amplifiable DNA were HPV positive. There was no change in HPV type over time and no statistically significant association between HPV type and disease aggressiveness. One novel, large-scale URR duplication was identified in an HPV type 11 isolate in the last of a series of six papillomas examined and the first from the bronchus. However, the duplication was not found in HPV type 11 isolates from the associated pulmonary carcinoma and its metastases in other organs. Three of 14 URR point mutations coincided with transcription factor binding sites, but there were no obvious associations with clinical course. Chi-square and multiple linear regression analyses of clinicopathological variables revealed early age at diagnosis (less than 4 years) as an independent predictor of aggressive disease (P < 0.001). A bimodal distribution of the age at diagnosis was noted, with peaks at 2 and 11 years of age. PMID- 9399502 TI - Comparison of enzyme immunoassay, PCR, and type-specific cDNA probe techniques for identification of group A rotavirus gene 4 types (P types). AB - This study was designed to evaluate three techniques most commonly used to identify the VP4 (P) types of human group A fecal rotaviruses. The techniques included PCR with nested primers and hybridization with PCR-generated probes (to determine the P genotypes). The results obtained by these genetic techniques were evaluated against those obtained by an enzyme immunoassay (EIA) incorporating neutralizing monoclonal antibodies (N-MAbs) reacting with three major human P serotypes (serotypes P1A, P1B, and P2A). The P types of the rotaviruses present in 102 fecal specimens were determined under code by each of the three assays. The specificity of each assay was evaluated against a "gold standard" putative P type (P serotype and genotype) deduced from knowledge of the VP7 (G) type and the origin of the fecal specimen. Overall comparison of the results showed respective sensitivities and specificities of 92 and 92% for reverse transcription-PCR, 80 and 99% for hybridization, and 73 and 91% for EIA with N-MAbs. The hybridization assay retained high sensitivity with specimens stored for > or = 10 years. Hybridization assays with nonradioactive probes are relatively inexpensive and are suited for use in developing countries. In summary, both genetic assays showed high sensitivities and specificities in assigning a P type to human fecal rotavirus strains. Further evaluation of the EIA with N-MAbs is required, together with incorporation of new N-MAbs for the detection of the additional P types detected in developing countries. PMID- 9399503 TI - Clinical utility of broth cultures of cerebrospinal fluid from patients at risk for shunt infections. AB - For patients with cerebrospinal fluid (CSF) shunts, culture of the CSF remains the most valuable tool in the evaluation of suspected shunt infections. To detect anaerobic Propionibacterium sp., a well-described cause of these infections, many clinical microbiology laboratories routinely employ a broth medium as an adjunct to solid media. The use of broth, however, creates a diagnostic dilemma since many contaminants also are isolated from broth cultures. Therefore, we retrospectively reviewed the records of 59 patients with CSF shunts in whom an organism was isolated from only broth cultures to assess their utility for the diagnosis of shunt infection. We found that no single clinical or laboratory parameter, including fever, leukocytosis, pleocytosis, or CSF protein and glucose, could reliably predict or exclude a shunt infection. Isolation of coagulase-negative staphylococci only in broth, in the absence of growth on solid media in concurrent or immediately preceding cultures, virtually always represented contamination. The isolation of Propionibacterium sp. from broth only usually represented contamination; however, infection could not be excluded without a repeated CSF culture, even in the absence of pleocytosis. We recommend that specific comments be appended to laboratory reports for isolates from CSF in broth only as an aid to the physician in interpreting the clinical importance of such isolates. PMID- 9399504 TI - Evaluation of six commercial kits for detection of human immunoglobulin M antibodies to Toxoplasma gondii. The FDA Toxoplasmosis Ad Hoc Working Group. AB - As a result of reports received by the Food and Drug Administration (FDA) of false-positive results obtained with FDA-cleared in vitro diagnostic kits for the detection of Toxoplasma-specific human immunoglobulin M (IgM) antibodies, an FDA sponsored evaluation of six kits was performed. A battery of 258 serum specimens, including 30 specimens drawn 1 to 5 months after initial Toxoplasma infection and 228 specimens from Toxoplasma IgG-positive individuals, Toxoplasma IgG-negative individuals, rheumatoid factor-positive persons, and persons determined to be Toxoplasma IgM positive by commercially available assays, was assembled, randomly assorted, and coded. The battery was tested at the FDA with six commercially available kits, at the Palo Alto Medical Foundation (PAMF) by the PAMF double sandwich IgM enzyme-linked immunosorbent assay (PAMF IgM ELISA), and at the Centers for Disease Control and Prevention (CDC) by the CDC EIA IgM. The results of the PAMF IgM ELISA that were obtained with the battery were considered to be the "gold standard" for this study; specificity rates were computed by considering the PAMF results to be 100% specific. Sensitivity and specificity rates were found to be as follows: CDC EIA IgM, 100 and 99.1%, respectively; Abbott IMx Toxo IgM, version 1, 100 and 77.5%, respectively; Abbott IMx Toxo IgM, version 2, 93.3 and 97.3%, respectively; Abbott Toxo-M EIA, 100 and 84.2%, respectively; BioMerieux Vitek VIDAS Toxo IgM, 100 and 98.6%, respectively; BioWhittaker Toxocap-M, 100 and 95.9%, respectively; Gull Toxo IgM, 97 and 85.6%, respectively; and Sanofi Diagnostics Pasteur Platelia Toxo IgM, 100 and 96.8%, respectively. Although the extent of false-positive reactions with these kits cannot be calculated because the study was retrospective and sample choices were biased, the results may be useful as an indicator of the relative specificities of these kits. PMID- 9399506 TI - Multicenter evaluation of the updated and extended API (RAPID) Coryne database 2.0. AB - In a multicenter study, 407 strains of coryneform bacteria were tested with the updated and extended API (RAPID) Coryne system with database 2.0 (bioMerieux, La Balme-les-Grottes, France) in order to evaluate the system's capability of identifying these bacteria. The design of the system was exactly the same as for the previous API (RAPID) Coryne strip with database 1.0, i.e., the 20 biochemical reactions covered were identical, but database 2.0 included both more taxa and additional differential tests. Three hundred ninety strains tested belonged to the 49 taxa covered by database 2.0, and 17 strains belonged to taxa not covered. Overall, the system correctly identified 90.5% of the strains belonging to taxa included, with additional tests needed for correct identification for 55.1% of all strains tested. Only 5.6% of all strains were not identified, and 3.8% were misidentified. Identification problems were observed in particular for Corynebacterium coyleae, Propionibacterium acnes, and Aureobacterium spp. The numerical profiles and corresponding identification results for the taxa not covered by the new database 2.0 were also given. In comparison to the results from published previous evaluations of the API (RAPID) Coryne database 1.0, more additional tests had to be performed with version 2.0 in order to completely identify the strains. This was the result of current changes in taxonomy and to provide for organisms described since the appearance of version 1.0. We conclude that the new API (RAPID) Coryne system 2.0 is a useful tool for identifying the diverse group of coryneform bacteria encountered in the routine clinical laboratory. PMID- 9399505 TI - Identification of Staphylococcus species and subspecies by the chaperonin 60 gene identification method and reverse checkerboard hybridization. AB - A previous study (S. H. Goh et al., J. Clin. Microbiol. 34:818-823, 1996) demonstrated that a 600-bp region of the chaperonin 60 (Cpn60) genes from various bacterial isolates could be amplified by PCR with a pair of degenerate primers and that the products could be used as species-specific probes for Staphylococcus aureus, S. epidermidis, S. haemolyticus, S. lugdunensis, S. saprophyticus, and S. schleiferi. To further validate the utility of bacterial Cpn60 genes as universal targets for bacterial identification (ID), reverse checkerboard chemiluminescent hybridization experiments were performed with DNA probes from 34 different Staphylococcus species and subspecies. With the exception of probes from the Cpn60 genes of S. intermedius and S. delphini, which cross hybridized, all were species specific. Two subspecies of both S. capitis and S. cohnii were differentiated from one another, while DNAs from the two S. schleiferi subspecies cross hybridized. When 40 known Staphylococcus isolates were tested in a blind experiment by the Cpn60 gene method, 36 strains, representing six species and one subspecies (S. sciuri, S. caseolyticus, S. hominis, S. warneri, S. hyicus, S. haemolyticus, and S. capitis subsp. ureolyticus), were correctly identified. DNA from the four remaining isolates, known to be S. hyicus bovine strains, failed to hybridize to DNA from the S. hyicus target strain or any other Staphylococcus species. However, DNAs from these S. hyicus isolates did cross hybridize with each other. New DNA sequence data and evidence from previous studies suggest some genetic divergence between the two groups of S. hyicus isolates. Our results demonstrate that this Cpn60 gene-based ID method has the potential to be a basic method for bacterial ID. Studies are in progress to further validate the utility of this Cpn60 gene system for ID of Staphylococcus and other genera, including those of slow-growing microorganisms. PMID- 9399507 TI - Evaluation of the MB/BacT system and comparison to the BACTEC 460 system and solid media for isolation of mycobacteria from clinical specimens. AB - The MB/BacT automated system is designed for the isolation of mycobacteria from clinical specimens. It utilizes a colorimetric sensor and reflected light to continuously monitor the CO2 concentration in the culture medium. We compared its performance to that of the BACTEC 12B media for the radiometric BACTEC 460 instrument and that of solid culture media. Respiratory specimens and urine samples were decontaminated with 4% NaOH. The vials of the two instruments were inoculated with 500 microl of sample and two solid egg-based media at 200 microl each. All vials were incubated at 37 degrees C for 6 weeks. A total of 1,078 specimens (633 respiratory specimens, 78 cerebrospinal fluid specimens, 177 other body fluid specimens, 87 urine specimens, and 103 other types of specimens) were cultured in parallel. Mycobacteria could be identified from 73 (6.8%) specimens: 67 M. tuberculosis, 3 M. kansasii, 1 M. xenopi, 1 M. terrae, and 1 mixed M. avium with M. scrofulaceum. Of these, 63 (86.3%) specimens were positive with the MB/BacT system, 67 (91.8%) were positive with the BACTEC 460 instrument, and 58 (79.5%) were positive with the two egg-based media. MB/BacT cultures were positive on average after 17.5 (+/-6.4) days, BACTEC cultures with a growth index of >20 (mean, 200) were positive after 14.3 (+/-8.2) days, and egg-based media were positive after 24.2 (+/-7.5) days. Microorganisms other than mycobacteria contaminated 46 (4.3%) MB/BacT cultures and 31 (2.9%) BACTEC cultures, which had to be discarded. The MB/BacT system is a well-automated system for the detection of M. tuberculosis in clinical specimens without using radioactive reagents. Further trials are required to determine whether it is suitable for the culture of nontuberculous mycobacteria. PMID- 9399508 TI - Randomly amplified polymorphic DNA PCR for comparison of Mycobacterium abscessus strains from nosocomial outbreaks. AB - Mycobacterium abscessus is an important cause of water-related nosocomial outbreaks or pseudo-outbreaks. Strain comparison has relied on pulsed-field gel electrophoresis (PFGE). Unfortunately, almost 50% of strains cannot be assessed by this method. We studied 118 strains of M. abscessus previously studied by PFGE by randomly amplified polymorphic DNA (RAPD) PCR, including isolates from eight nosocomial outbreaks. Ten random primers were evaluated by using DNA prepared by boiling or phenol-chloroform extraction. Both DNA preparations gave the same grouping of isolates for three outbreaks compared to the groupings obtained by PFGE. Five outbreaks due to M. abscessus which gave broken DNA by PFGE gave evaluable patterns when studied by RAPD-PCR, with isolate clustering being consistent with that from other laboratory and epidemiologic data. The patterns were highly method dependent, strain comparison required the use of multiple primers, and the method worked best with purified DNA and by using strains for comparison on the same gel. We propose categories of indistinguishable, different, and inconclusive when comparing strains by RAPD-PCR. This study demonstrates that RAPD-PCR can be used for genetic comparison of M. abscessus strains, including strains which cannot be compared by PFGE, but the potential for misinterpretation is greater than that by PFGE. PMID- 9399509 TI - Identification of pathogenic Leptospira genospecies by continuous monitoring of fluorogenic hybridization probes during rapid-cycle PCR. AB - Partial sequences of 23S rRNA gene PCR products from 23 strains of 6 pathogenic Leptospira genospecies and from 8 strains of the saprophytic Leptospira biflexa were determined. Sequence analyses enabled Leptospira genus-specific amplification primers and pathogen-specific fluorogenic adjacent hybridization probes to be designed and synthesized. A PCR protocol was developed in which changes in fluorescence emission resulting from specific annealing of fluorogenic adjacent hybridization probes to the target DNA were continuously monitored. Nine strains of the pathogenic Leptospira genospecies could be differentiated from Leptonema illini, Escherichia coli, and eight strains of Leptospira biflexa. The PCR method was rapid, requiring 18 min for the completion of 45 cycles. It was also simple and flexible, as DNA templates prepared by four different methods, including the simple boiling method, could be used without adverse effects. Two hundred copies of target, equivalent to 100 cells, could be detected. PMID- 9399510 TI - Reversed passive latex agglutination assay for detection of toxigenic Corynebacterium diphtheriae. AB - A reversed passive latex agglutination (RPLA) assay for determining the toxigenicity of Corynebacterium diphtheriae is presented. Rabbit antitoxin antiserum was raised by using commercially available diphtheria toxoid. This antiserum reacted with the diphtheria toxin when the culture supernatant was assayed by Western blotting, and it did not cross-react with other extracellular antigens. Affinity-purified antibodies for latex sensitization were obtained by using a Hi Trap N-hydroxysuccinimide-activated column. Demonstration of toxin in five of seven clinical isolates was in accordance with the PCR assay and the Vero cell cytotoxicity test. Culture of the bacteria for 6 h was sufficient for toxin production, and an additional 6 h was needed to observe latex agglutination. Therefore, diphtheria toxin can be detected in 12 h by this method. The lowest concentration of diphtheria toxin detectable by the RPLA assay was about 5 ng/ml. The RPLA assay can provide a convenient and reliable method for laboratories involved in the identification of toxinogenic corynebacteria. PMID- 9399511 TI - Emergence of a unique O3:K6 clone of Vibrio parahaemolyticus in Calcutta, India, and isolation of strains from the same clonal group from Southeast Asian travelers arriving in Japan. AB - Active surveillance of Vibrio parahaemolyticus infection among hospitalized patients in Calcutta, India, was initiated in January 1994. The incidence of cases of V. parahaemolyticus infection suddenly increased in February 1996 and has remained high since then. One hundred thirty-four strains of V. parahaemolyticus isolated from January 1994 to August 1996 were examined for serovar, the presence of the thermostable direct hemolysin gene (tdh) and tdh related hemolysin genes (trh1 and trh2), production of urease, and antibiogram. Strains of the O3:K6 serovar appeared for the first time in February 1996. The O3:K6 serovar strains accounted for 50 to 80% of the strains isolated during the high-incidence period (February to August 1996). All of the serovar O3:K6 strains carried the tdh gene but not the trh genes and did not produce urease. All of the isolates except two were sensitive to all of the antibiotics tested. These and the results of analysis by an arbitrarily primed PCR method indicated that the O3:K6 serovar strains belong to a unique clone. When the O3:K6 serovar strains, isolated from travelers arriving in Japan from Southeast Asian countries, were compared by the arbitrarily primed PCR method, the strains isolated between 1982 and 1993 were distinct from Calcutta O3:K6 while the strains isolated in 1995 and 1996 were indistinguishable from the Calcutta O3:K6 strains. The results suggest that this unique O3:K6 clone may have become prevalent not only in Calcutta but also in Southeast Asian countries very recently. Not only the O3:K6 strains but also the non-O3:K6, tdh-bearing strains isolated in 1996 produced thermostable direct hemolysin at high levels, and thus the level of hemolysin produced does not appear to have influenced the high incidence of serovar O3:K6 strains. PMID- 9399512 TI - Determination of hepatitis C virus genotypes in the United States by cleavase fragment length polymorphism analysis. AB - We describe the application of a new DNA-scanning method, which has been termed Cleavase Fragment Length Polymorphism (CFLP; Third Wave Technologies, Inc., Madison, Wis.), for the determination of the genotype of hepatitis C virus (HCV). CFLP analysis results in the generation of structural fingerprints that allow discrimination of different DNA sequences. We analyzed 251-bp cDNA products generated by reverse transcription-PCR of the well-conserved 5'-noncoding region of HCV. We determined the genotypes of 87 samples by DNA sequencing and found isolates representing 98% of the types typically encountered in the United States, i.e., types 1a, 1b, 2a/c, 2b, 3a, and 4. Blinded CFLP analysis of these samples was 100% concordant with DNA sequencing results, such that closely related genotypes yielded patterns with strong familial resemblance whereas more divergent sequences yielded patterns with pronounced dissimilarities. In each case, the aggregate pattern was indicative of genotypic grouping, while finer changes suggested subgenotypic differences. We also assessed the reproducibility of CFLP analysis in HCV genotyping by analyzing three distinct isolates belonging to a single subtype. These three isolates yielded indistinguishable CFLP patterns, as did replicate analysis of a single isolate. This study demonstrates the suitability of this technology for HCV genotyping and suggests that it may provide a low-cost, high-throughput alternative to DNA sequencing or other, more costly or cumbersome genotyping approaches. PMID- 9399513 TI - Outbreak of Shigella sonnei in a clinical microbiology laboratory. AB - Laboratory technologists (22%) developed infections with Shigella sonnei. The isolates had the same antibiogram and pulse-field gel electrophoresis pattern as an unknown isolate handled by a laboratory student. Covering faucet handles with paper towels during hand washing in the laboratory was protective. No further cases occurred after the laboratory was cleaned with a phenolic agent and a handle-free faucet was installed. PMID- 9399514 TI - Clinical and epidemiological significance of enterococci intrinsically resistant to vancomycin (possessing the vanC genotype) AB - Constitutive low-level vancomycin resistance is found intrinsically in certain enterococcal species and is encoded by vanC ligase genes. These intrinsically vancomycin-resistant enterococci (VRE) will be referred to as VANC VRE. A prospective study to determine the clinical and epidemiologic significance of VANC VRE was conducted. VANC VRE were recovered from the stools of 34 of 601 (5.7%) patients, a rate similar to that obtained for the stools of 100 outpatients in the community (5%). VANC VRE were also isolated from the nonstool specimens of 9 of 538 patients (1.7%), including two patients with bacteremia. No VRE of the vanA or vanB genotypes were detected in nonstool specimens. Eighty-two hospital contacts of the first 23 patients found to be colonized or infected with VANC VRE were screened, and 6 contacts were found to be gastrointestinal carriers of VANC VRE. However, typing of isolates from these 6 contacts by pulsed-field gel electrophoresis with SmaI showed the isolates to be unique and different from those recovered from the index patients. In fact, all VANC VRE isolates from different patients in this study were unique. A case-control study with patients who were negative when screened for VANC VRE as controls failed to identify any risk factor associated with colonization or infection with this organism. VANC VRE were infrequently recovered from clinical specimens but were occasionally found as part of the normal stool flora. Since no transmission between patients was documented, additional isolation procedures may not be necessary for patients colonized or infected with VANC VRE. PMID- 9399515 TI - Comparison of use of phenotypic and genotypic characteristics for identification of species of the anamorph genus Candida and related teleomorph yeast species. AB - A total of 49 type and neotype isolates and 32 clinical isolates of the anamorph genus Candida and related teleomorph genera were obtained from different culture collections and clinical laboratories. Isolates were subjected to two phenotypic methods of identification, Vitek yeast biochemical card (YBC) and API ID 32C, both based on carbohydrate assimilation, and one genotypic method, PCR fingerprinting, based on the detection of DNA polymorphisms between minisatellite specific sequences with the primer M13 (5' GAGGGTGGCGGTTCT 3'). The correct identification of a strain at the Centraalbureau voor Schimmelcultures was used as the gold standard for the identification of an isolate. When the study was restricted to species included in the respective biochemical databases, the Vitek YBC and API ID 32C systems performed adequately with positive identification rates of 87.3 and 76.8%, respectively. When uncommon species were added to the study, several of which are not included in the databases, the identification efficiencies were 76.5 and 77.5%, respectively. By comparison, all isolates were correctly identified by PCR fingerprinting, with 63 reference species profiles in the databank. Sufficient polymorphisms among the total set of banding patterns were observed, with adequate similarity in the major patterns obtained from a given species, to allow each isolate to be assigned unambiguously to a particular species. In addition, variations in minor bands allowed for differentiation to the strain level. PCR fingerprinting was found to be rapid, reproducible, and more cost-effective than either biochemical approach. Our results provide reference laboratories with an improved identification method for yeasts based on genotypic rather than phenotypic markers. PMID- 9399516 TI - Fecal microflora in a patient with short-bowel syndrome and identification of dominant lactobacilli. AB - Fecal microflora and lactate concentrations in blood and feces obtained from a patient (a 5 year-old boy) with short-bowel syndrome (SBS) were compared during acidosis to results for the normal condition (no SBS symptoms). The taxonomical position of the lactobacilli found predominantly in the feces sample obtained 2 days before the fifth attack was also studied. The D-lactate level in serum obtained 1 day after the fourth attack was 10-fold higher than that for the normal condition, although there was not a great difference in L-lactate levels. D-Lactate (3.91 mM) and L-lactate (2.86 mM) were also detected in the feces samples collected 2 days before the fifth attack, while no lactate was detected in the feces sample for the normal condition. The counts of total fecal bacteria, especially anaerobic bacteria such as members of the family Bacteroidaceae, were found to be low. The counts of lactobacilli and the total population of lactobacilli relative to total fecal bacteria in the feces 2 days before the fifth attack (40.4%) were extremely high. In this case, a majority of the lactobacilli were D-lactate producers as determined by homolactic fermentation. These lactobacilli were identified as Lactobacillus delbrueckii subsp. lactis. The percentages of bifidobacteria relative to total fecal bacteria in feces samples obtained both 2 days before the fifth attack (50.9%) and for normal condition (61.9%) were also high, although these bacteria were L-lactate producers. In the feces samples for the normal condition, the D-lactate producers decreased to less than 10(9) per g, while the counts of L- or DL-lactate producers were 100-fold higher than the numbers in feces samples obtained 2 days before the fifth attack. These results suggested that an increase in the level of D-lactate producers, such as L. delbrueckii subsp. lactis, in the colon may be associated with the clinical expression of metabolic acidosis. PMID- 9399517 TI - Evaluation of the BBL Crystal Anaerobe identification system. AB - The BBL Crystal Anaerobe (ANR) identification system was evaluated, and the results were compared with those from conventional anaerobic methods. We tested 322 clinically significant anaerobic bacteria according to the manufacturer's instructions. The system identified correctly 286 of 322 (88.8%) of the anaerobic bacteria tested. Of these, 263 of 322 (81.7%) were identified correctly on initial testing and 49 were identified correctly only to the genus level; on repeat testing, 23 of 49 (46.9%) were identified correctly to both the genus and the species levels. A total of 26 (8.5%) were misidentified at the species level, and 10 (3.1%) were not identified. Performance characteristics for individual strains varied. The system correctly identified all tested strains of Campylobacter, Desulfomonas, Desulfovibrio, Leptotrichia, Mobiluncus, Peptostreptococcus, Porphyromonas, Provetella, Propionibacterium, Tisierella, and Veillonella and 36 of 37 (97.3%) Actinomyces strains, 42 of 46 (91.3%) B. fragilis group strains, 79 of 103 (76.7%) Clostridium strains, (however, the system failed to identify any of the 7 C. innocuum and 9 C. tetani strains tested), and 8 of 15 (53.3%) Bacteroides strains. This system was easy to use, did not involve the addition of reagents, and was faster than conventional anaerobic procedures. It would be a useful addition to the anaerobe laboratory of most hospitals. PMID- 9399518 TI - Highly sensitive single-step PCR protocol for diagnosis and monitoring of human cytomegalovirus infection in renal transplant recipients. AB - A multiplex, single-step PCR protocol for the detection of human cytomegalovirus (HCMV) DNA is described. The protocol amplifies regions of the viral LA and IE genes and employs elevated temperatures for both reagent mixing and primer annealing together with product detection by silver staining on polyacrylamide gels. This assay detects one to five HCMV genomes in clinical samples containing up to 100 ng of human DNA, a level of sensitivity equivalent to that of more complex assays involving either nested PCR or postamplification hybridization. As well as being of importance in clinical situations where high-sensitivity qualitative diagnosis is required, this assay is also applicable to the monitoring of HCMV infection in renal transplant recipients. Due to its multiplex format the assay provides quantitative information, in that samples from which a single target is amplified contain on average sevenfold fewer viral genomes per 10(6) leukocytes than those from which both targets are amplified. When weekly blood leukocyte DNA preparations from renal transplant patients were assayed, findings of three consecutive tests in which both HCMV targets were amplified were highly indicative of patients who had developed very high loads of HCMV (100% sensitivity, 88% specificity). We thus show that the same simple PCR assay which permits highly sensitive HCMV diagnosis can also be used for the efficient identification of transplant recipients at risk of clinically significant infection. PMID- 9399519 TI - Disk diffusion test interpretive criteria and quality control recommendations for testing linezolid (U-100766) and eperezolid (U-100592) with commercially prepared reagents. AB - Two new oxazolidinones were tested to determine interpretive susceptibility testing criteria for MIC and disk diffusion methods. Commercial lots of linezolid (formerly U-100766) and eperezolid (formerly U-100592) disks containing 30 microg of drug were tested against 728 isolates of bacteria with defined mechanisms of resistance. Results from linezolid were highlighted because of its choice for clinical development. By using preliminary pharmacokinetic data, a tentative susceptibility breakpoint of < or = 4 microg/ml was selected. Corresponding breakpoint zone diameters for linezolid were > or = 21 mm (< or = 4 microg/ml) for susceptibility and < or = 17 mm (> or = 16 microg/ml) for resistance. Regression statistics demonstrated a high correlation coefficient (r > or = 0.98), and absolute categorical agreement between methods was obtained, when staphylococci and enterococci were tested with the cited criteria. When Streptococcus spp. (including S. pneumoniae) were tested, only the susceptibility breakpoint was suggested. Quality control (QC) guidelines for linezolid disk diffusion tests were established by a multilaboratory trial as follows: 27 to 31 mm for Staphylococcus aureus ATCC 25923 and 28 to 34 mm for S. pneumoniae ATCC 49619. More than 95% of all QC results were within these proposed ranges. Although not advanced to clinical trials, eperezolid demonstrated potency comparable to that of linezolid and had identical interpretive testing criteria. These preliminary interpretive criteria and QC limits (accepted by the National Committee for Clinical Laboratory Standards) should be applied to linezolid tests during the clinical-trial phases of oxazolidinone drug development in order to ensure test accuracy and reproducibility. PMID- 9399520 TI - A new agent of mycobacterial lymphadenitis in children: Mycobacterium heidelbergense sp. nov. AB - Nontuberculous mycobacterial lymphadenitis presents an increasing clinical problem in immunocompetent young children. A slowly growing, nonphotochromogenic mycobacterium was recovered twice (isolates 2553/91 and 2554/91) from the lymphatic tissue of a child with recurrent cervical lymphadenitis. It could be differentiated biochemically from described Mycobacterium species, although it most closely resembled Mycobacterium malmoense by thin-layer chromatography and high-performance liquid chromatography of mycolic acids. A striking characteristic of the isolate was its high degree of susceptibility to antituberculous drugs in vitro, including isoniazid. Direct determination of the 16S rRNA gene sequence revealed a unique sequence and positioned the strain phylogenetically on a branch separate from M. malmoense within a group of slowly growing mycobacteria that show a high degree of similarity to M. simiae at the 16S rRNA gene level. Despite 99.6% sequence identity with M. simiae at the 16S rRNA gene level, DNA-DNA hybridization studies (hydroxyapatite method) demonstrated DNA relatedness of less than 40%. We conclude that this organism is a new species for which we propose the name M. heidelbergense. A culture of the type strain, strain 2554/91, has been deposited in the American Type Culture Collection as strain ATCC 51253. PMID- 9399521 TI - Laboratory methods for detection of Chlamydia trachomatis: survey of laboratories in Washington State. AB - The last decade has witnessed the development of a wide variety of diagnostic tests for Chlamydia trachomatis. In order to determine what laboratory methods are being used to detect C. trachomatis infections in Washington State and to identify factors influencing test selection, between April 1995 and October 1995 we conducted a mailed questionnaire survey of all 112 laboratories certified to do chlamydia testing. Of these, 20 had discontinued testing for C. trachomatis, and responses were obtained from 89 (97%) of the remaining 92 laboratories. Surprisingly, 38 (43%) of the 89 laboratories used rapid tests such as Clearview and Surecell, making such tests the most commonly used laboratory tests. Laboratories which used rapid tests had lower test volumes, less experience performing tests for C. trachomatis, less frequent attendance at professional meetings, and greater reliance on manufacturers for information compared with laboratories which used other methods. Confirmation of non-culture-positive results was provided by 28 (34%) of the 82 laboratories doing non-culture-based tests. Forty-one (47%) of 88 laboratories reported having compared their method with another method. Test volume was the strongest predictor of laboratories which confirmed positive non-culture-based test results and which had performed a laboratory comparison of methods. We conclude that rapid tests for C. trachomatis are often being used inappropriately and that efforts are needed to improve effective implementation and quality assurance of laboratory testing for C. trachomatis. PMID- 9399522 TI - Confirmation of suspicious cases of meningococcal meningitis by PCR and enzyme linked immunosorbent assay. AB - A significant problem in efficacy trials of meningococcal vaccines has been accurate identification of all cases of meningococcal disease that occur in study populations. The accuracy of case determination would be improved by utilizing methods which confirm or disprove suspicious cases of meningococcal disease that are culture negative. A collection of serum and cerebrospinal fluid (CSF) samples from a meningococcal vaccine field trial performed in Iquique, Chile, were utilized to assess the status of patients for whom cultures, Gram stains, and clinical evaluations for meningococcal disease were available. Nested PCRs (nPCRs) for amplification of Neisseria meningitidis DNA in CSF samples and enzyme linked immunosorbent assays (ELISAs) for quantification of serum immunoglobulin G antibodies specific for N. meningitidis were used in combination to confirm or eliminate cases classified by physicians as suspicious for meningococcal disease. Samples from 12 of 79 patients suspected of having meningococcal meningitis tested positive by both methods; specimens from 61 of the 79 were negative by both methods; and samples from 6 patients yielded ambiguous results, and these cases remained unconfirmed. Direct sequence analysis of amplified DNA from patients suspected of having meningococcal disease confirmed that 2 of the 12 newly confirmed cases were not attributable to the typical epidemic strain (B:15:P1.[7],3) while the others were due to the epidemic strain. A combination of nPCR and ELISA reduced the number of suspicious cases in this study from 79 to 6, thereby improving the potential for assessment of vaccine efficacy. Molecular identification by nPCR in conjunction with immunological assessment of patient response could be considered diagnostic of disease in future testing of meningococcal vaccines to improve efficacy analyses. PMID- 9399523 TI - Characterization of group A Streptococcus strains recovered from Mexican children with pharyngitis by automated DNA sequencing of virulence-related genes: unexpectedly large variation in the gene (sic) encoding a complement-inhibiting protein. AB - Sequence variation was studied in several target genes in 54 strains of group A Streptococcus (GAS) cultured from children with pharyngitis in Mexico City. Although 16 distinct emm alleles were identified, only 4 had not been previously described. Virtually all bacteria (31 of 33 [94%] with the streptococcal pyrogenic exotoxin gene (speA) had emm1-related, emm3, or emm6 alleles. The gene (sic) encoding an extracellular GAS protein that inhibits complement function was unusually variable among isolates with the emm1 family of alleles, with a total of seven variants identified. The data suggest that many GAS strains infecting Mexican children are genetically similar to organisms commonly encountered in the United States and western Europe. Sequence variation in the sic gene is useful for rapid differentiation among GAS isolates with the emm1 family of alleles. PMID- 9399524 TI - Specific identification of Mycobacterium tuberculosis with the luciferase reporter mycobacteriophage: use of p-nitro-alpha-acetylamino-beta-hydroxy propiophenone. AB - We have previously described a luciferase reporter mycobacteriophage (LRP) assay that can detect Mycobacterium tuberculosis and characterize mycobacterial drug susceptibility patterns within 24 to 48 h in positive cultures. One drawback of this LRP protocol is the ability of the recombinant mycobacteriophage phAE40 to infect a variety of Mycobacterium species, thus limiting its specificity for the detection of M. tuberculosis. In this study, we have (i) explored the host range of phAE40, (ii) developed a modified LRP assay that exploits the selective inhibitory effect of the compound p-nitro-alpha-acetylamino-beta-hydroxy propiophenone (NAP) against members of the M. tuberculosis complex to differentiate between the tubercle bacillus and other mycobacterial species, and (iii) tested over 300 samples, including primary clinical isolates and drug resistant strains of M. tuberculosis, demonstrating the ability of the NAP modified LRP assay to identify M. tuberculosis complex organisms with high degrees of sensitivity and specificity. PMID- 9399525 TI - Conditionally replicating luciferase reporter phages: improved sensitivity for rapid detection and assessment of drug susceptibility of Mycobacterium tuberculosis. AB - TM4 is a lytic mycobacteriophage which infects mycobacteria of clinical importance. A luciferase reporter phage, phAE40, has been constructed from TM4 and was previously shown to be useful for the rapid detection and drug susceptibility testing of Mycobacterium tuberculosis. However, the lytic nature of the phage results in a loss of detectable light output and limits the sensitivity of detection. We describe several strategies aimed at improving the luciferase activity generated by TM4 luciferase phages, including (i) varying the position of the luciferase gene in the phage genome, (ii) isolating host-range mutants of the phage, and (iii) introducing temperature-sensitive mutations in the phage such that it will not replicate at the infecting temperature. Several new phages generated by these methods show increased intensity of luciferase production compared to the first-generation reporter phage phAE40, and one phage, phAE88, also demonstrates an enhanced duration of luciferase activity. This has allowed the detection of as few as 120 BCG cells and the determination of drug susceptibilities of M. tuberculosis in as little as 1 day. PMID- 9399526 TI - Use of the BioMerieux ID 32C yeast identification system for identification of aerobic actinomycetes of medical importance. AB - The BioMerieux ID 32C Yeast Identification System was examined to determine its usefulness as a rapid method for the identification of medically important aerobic actinomycetes. More than 290 strains were tested by this method and the results were compared to those obtained by conventional methods. It was found that aerobic actinomycetes could be differentiated to species level in 7 days by the ID 32C system. PMID- 9399527 TI - Human granulocytic ehrlichiosis in southern Germany: increased seroprevalence in high-risk groups. AB - To date, human granulocytic ehrlichiosis (HGE), the causative agent of which is likely transmitted by ticks in the Ixodes ricinus-Ixodes persulcatus complex, has not been diagnosed with certainty in patients outside the United States. The presence of a closely related vector tick, I. ricinus, as well as the occurrence of similar Ehrlichia spp. of veterinary importance, suggests that this disease is likely to be present in Europe. The aim of the present study was to compare the prevalence of antibodies against the HGE agent in sera collected from patients in groups at high risk for exposure to I. ricinus with that of a control population. Risk groups consisted of 150 forestry workers and 105 patients with an established diagnosis of Lyme disease. The control group was 103 healthy blood donors without a history of tick bites. We used a patient isolate of the HGE agent from Minnesota (J. L. Goodman, C. Nelson, B. Vitale, J. E. Madigan, J. S. Dumler, T. J. Kurtti, and U. G. Munderloh, N. Engl. J. Med. 334:209-215, 1996) propagated in HL60 cells as the source of antigen for a specific immunofluorescence assay (IFA). Elevated IFA titers (> or = 1:80) were present in 21 of 150 (14%) serum samples from forestry workers and in 12 of 105 (11.4%) serum samples from Lyme disease patients, but in only 2 of 103 (1.9%) serum samples from blood donors (P < or = 0.01 for either of the at-risk groups versus blood donors). The results of this study suggest that the HGE agent or a closely related organism exists in southern Germany and that seroconversion to it is common among groups exposed to Ixodes ticks. Final proof that HGE occurs in Germany will require the isolation of the causative agent from patients. HGE should be considered in the differential diagnosis of febrile illnesses in individuals exposed to Ixodes ticks in Europe as well as in North America. PMID- 9399528 TI - Direct detection of Mycobacterium tuberculosis complex in clinical samples from patients in Norway by ligase chain reaction. AB - Our aim was to investigate the use of DNA amplification with the ligase chain reaction (LCR) for detection of the Mycobacterium tuberculosis complex directly in human clinical specimens. The LCR assay employed was the Abbott LCx MTB Assay, which uses the gene encoding protein antigen b as the target template. Four hundred eighty-two samples from 457 patients in one clinical microbiology laboratory in Norway were processed by routine culture analysis (BACTEC culture), direct microscopy (Ziehl-Neelsen staining) and LCR. Of the 118 specimens containing cultivable M. tuberculosis, 106 (90.6%) were detected by LCR. Among the 364 culture-negative specimens, 356 samples were negative also by LCR and 8 (1.6%) were positive by LCR. In five of the eight LCR-positive and culture negative samples, another sample from the same patient was M. tuberculosis culture positive and/or the patient had symptoms of tuberculosis. In comparison with culture, the sensitivity of LCR was 96.7% for smear-positive samples and 72.0% for smear-negative samples, respectively. For all samples combined, the sensitivity, specificity, and positive and negative predictive values were 90.2, 99.2, 97.4, and 96.7%, respectively. Challenging the M. tuberculosis LCR test with DNAs and cultures from strains of Mycobacterium ulcerans and Mycobacterium marinum, which are the mycobacterial species most closely related to the M. tuberculosis complex, resulted in all-negative test results. The sensitivity, specificity, and positive and negative predictive values of BACTEC culture in comparison with the LCR test and clinical criteria were 95.9, 100, 100, and 98.6%, respectively. A certain prioritization of samples subjected to the LCR assay should be based on clinical indications and risks with regard to infection transmission and patient isolation policy. More automation and lower expenses are generally desired for nucleic acid amplification kits. However, this M. tuberculosis LCR assay represents a valuable tool in routine mycobacterial diagnostics. PMID- 9399529 TI - Comparison of PCR, culture, and histopathology for diagnosis of tuberculous pericarditis. AB - Nucleic acid amplification techniques for the diagnosis of tuberculosis (TB) are rapidly being developed. Scant work, however, has focused on pericardial TB. Using cryopreserved specimens from a prior study of pericarditis, we compared PCR to culture and histopathology for the diagnosis of tuberculous pericarditis in 36 specimens of pericardial fluid and 19 specimens of pericardial tissue from 20 patients. Fluid and tissue were cultured on Lowenstein-Jensen and Middlebrook solid media and in BACTEC radiometric broth. Tissue specimens were stained with hematoxylin-eosin, Ziehl-Neelsen, auramine O, and Kinyoun stains and were examined for granuloma formation and acid-fast bacilli. PCR was performed with both fluid and tissue with IS6110-based primers specific for the Mycobacterium tuberculosis complex by published methods. Sixteen of the 20 patients had tuberculous pericarditis and 4 patients had other diagnoses. TB was correctly diagnosed by culture in 15 (93%) patients, by PCR in 13 (81%) patients, and by histology in 13 of 15 (87%) patients. PCR gave one false-positive result for a patient with Staphylococcus aureus pericarditis. Considering the individual specimens as the unit of analysis, M. tuberculosis was identified by culture in 30 of 43 specimens (70%) from patients with tuberculous pericarditis and by PCR in 14 of 28 specimens (50%) from patients with tuberculous pericarditis (P > 0.15). The sensitivity of PCR was higher with tissue specimens (12 of 15; 80%) than with fluid specimens (2 of 13; 15%; P = 0.002). In conclusion, the overall accuracy of PCR approached the results of conventional methods, although PCR was much faster. Therefore, PCR merits further development in this regard. The sensitivity of PCR with pericardial fluid was poor, and false-positive results with PCR remain a concern. PMID- 9399530 TI - Comparison of agar dilution, broth microdilution, E-test, disk diffusion, and automated Vitek methods for testing susceptibilities of Enterococcus spp. to vancomycin. AB - An evaluation was undertaken to determine the optimal method for testing the susceptibilities of 100 clinical isolates and two reference strains of Enterococcus spp. to vancomycin in vitro. Six testing methods were studied by using the following media and incubation times: agar screen with the Synergy Quad Plate (Remel, Lenexa, Kans.), an in-house-prepared brain heart infusion (BHI) agar plate, and an in-house-prepared Mueller-Hinton (MH) agar plate, all incubated for 24 or 48 h; broth microdilution (Sensititre Just One Strip; AccuMed International, Inc., West Lake, Ohio) with BHI or cation-adjusted MH broth incubated for 24 or 48 h; agar dilution with BHI or MH agar incubated for 24 or 48 h; epsilometer test (E test; AB BioDisk, Solna, Sweden) with BHI or MH agar incubated for 24 or 48 h; disk diffusion with BHI or MH agar incubated for 24 or 48 h; and the automated Vitek method with the gram-positive susceptibility Staphylococcus aureus card and R02.03 software (bioMerieux, Inc., Hazelwood, Mo.). Growth failures occurred with MH media (n = 6) but not with BHI media. One growth failure occurred with the Vitek method. Results for each testing method for each Enterococcus strain were interpreted as susceptible, intermediate, or resistant according to current National Committee for Clinical Laboratory Standards (NCCLS) criteria and compared to the vancomycin resistance genotype (i.e., vanA, vanB, vanC-1, or vanC-2/3). For all methods, extension of the incubation time from 24 h to 48 h either produced no difference in the results or gave poorer results. The following methods produced no very major or major interpretive errors: broth microdilution with BHI media incubated for 24 h, agar dilution with BHI media incubated for 24 or 48 h, and E test with BHI media incubated for 24 or 48 h. Unacceptable frequencies of very major errors (> 1%) occurred with all methods for which MH media were used. Minor interpretive errors were frequent with all methods. These minor interpretive errors also occurred most frequently with Enterococcus strains with vanC genes, which encoded low level vancomycin resistance (MIC < or = 8 microg/ml), as opposed to Enterococcus strains which possessed vanA or vanB genes, which encoded higher-level vancomycin resistance (MIC > or = 64 microg/ml). Modification of NCCLS breakpoints, especially for motile Enterococcus spp. (E. casseliflavus, E. flavescens, and E. gallinarum), may resolve this problem; however, in the current study, one E. faecalis strain and one E. faecium strain carried only the vanC gene. The agar screen method may also require reformulation. The current agar screen plate contains 6 microg of vancomycin per ml, which may not detect all low-level resistance associated with vanC genotypes. Nevertheless, the clinical significance of this low-level vancomycin resistance remains unknown. PMID- 9399532 TI - Evaluation of Vitek GNI+ and Becton Dickinson Microbiology Systems Crystal E/NF identification systems for identification of members of the family Enterobacteriaceae and other gram-negative, glucose-fermenting and non-glucose fermenting bacilli. AB - We evaluated the Vitek GNI+ and Becton Dickinson Crystal E/NF identification systems for their ability to accurately identify 619 and 626 strains, respectively, of members of the family Enterobacteriaceae and other glucose fermenting and non-glucose-fermenting gram-negative rods. All strains tested were taken from a stock collection and passed three times on 5% sheep blood agar prior to testing. These strains represented a more rigorous challenge to both systems than one resulting from the testing of consecutive clinical isolates. Testing with both systems was done according to the manufacturers' instructions, and tests were repeated in duplicate when errors occurred. Vitek version 5.01 and Crystal version 3.0 softwares were used for identifications. The identification results from each system were compared with identifications previously determined with reference biochemicals. At the completion of the appropriate incubation period, the GNI+ and Crystal systems correctly identified 80.1 and 71.1% of the total isolates, respectively. After additional tests suggested by the software programs were completed, the GNI+ had an accuracy of 87.6% and the Crystal system's accuracy had improved to 87.9%. The error rates for the GNI+ and Crystal systems were 6.5 and 5.3%, respectively. A report of "no identification" was given for 6.0 and 6.9% of the isolates, respectively, and was associated with no particular organism group. One isolate each of Acinetobacter lwoffii and Vibrio alginolyticus would not grow in the Vitek card. The average times to detection for correct enteric identifications in the GNI+ system were 4.1 and 6.8 h for nonenteric identifications, while the Crystal results were routinely read at 18 h. We conclude that there was no significant difference (P > 0.05) between the results of the GNI+ card and those of the Crystal E/NF system after additional testing was performed with the group of organisms tested, but the overall accuracy for both systems in this study was below 90%. PMID- 9399531 TI - Evaluation of serological methods for diagnosis of Puumala hantavirus infection (nephropathia epidemica). AB - Nephropathia epidemica (NE), Puumala (PUU) virus infection, is a febrile disease which is commonly associated with acute renal impairment. To differentiate NE from other acute febrile illnesses, a rapid and reliable serological diagnosis is important, and a number of different protocols have recently been introduced. In the present report we describe a comparative evaluation of six PUU virus immunoglobulin M (IgM) and seven IgG enzyme-linked immunosorbent assay (ELISA) protocols based on native, Escherichia coli-expressed, or baculovirus-expressed nucleocapsid protein (N). Neutralization and immunofluorescence assays were included for comparison. Equally high sensitivities and specificities were obtained with three mu-capture-based IgM ELISAs using native, baculovirus expressed, and E. coli-expressed N antigens, respectively, and by an ELISA based on purified E. coli-expressed full-length N adsorbed to solid phase. The assays based on truncated amino-terminal N proteins, including a commercially available PUU virus IgM ELISA, all showed lower sensitivities. For detection of PUU virus specific IgG, ELISAs based on monoclonal antibody-captured native or baculovirus expressed N antigens showed optimal sensitivities and specificities, while the assays based on E. coli-expressed N did not detect all PUU virus IgG-positive serum samples. A commercially available PUU virus IgG ELISA based on E. coli expressed amino-terminal N showed a significantly lower sensitivity than those of all other IgG assays. PMID- 9399533 TI - Incidence and risk factors for hepatitis C among injection drug users in Baltimore, Maryland. AB - Between 1988 and 1996, the incidence of and risk factors for hepatitis C virus (HCV) infection were studied in a cohort of injection drug users in Baltimore, Maryland. By second-generation antibody testing of stored serum samples, 142 participants were found to be susceptible to HCV at the time they entered the study. After a median follow-up of 6.5 years, 43 participants (30.3%) developed antibodies to HCV (anti-HCV). The overall incidence was 6.4 cases per 100 person years, but a substantial decline in the annual incidence rate was observed after the first 2 years (1988 to 1990, 13.4/100 person-years; 1991 to 1996, 2.3/100 person-years [P = 0.0001 for trend]). Participants who acknowledged active drug use, especially those who acknowledged frequent use and sharing of drug paraphernalia, were at increased risk of HCV infection. However, high-risk sexual practices were not associated with HCV seroconversion. Efforts to reduce HCV infection must be focused on curbing drug use and especially on the sharing of needles and drug paraphernalia. PMID- 9399534 TI - Use of monoclonal antibodies in diagnosis of paracoccidioidomycosis: new strategies for detection of circulating antigens. AB - The precise diagnosis of paracoccidioidomycosis, in most cases, is established by direct methods and indirect immunological tests. The latter method is reliant on the identification of the host's humoral responses, which are usually impaired or absent in patients with severe juvenile forms of the disease and in immunocompromised patients. Determining disease activity or assessing treatment responses by measuring antibody levels is difficult, since antibody titer may remain elevated or persist at stationary levels, even in the presence of clinical improvement. Consequently, there is a need for alternative tests aimed at the identification of circulating antigens. A modification of the standard hybridoma production method was used to raise a panel of murine monoclonal antibodies (MAbs) against the yeast form of Paracoccidioides brasiliensis. Of these, MAb PIB, directed against an 87-kDa determinant, was used to develop an inhibition ELISA (inh-ELISA) capable of detecting as little as 5.8 ng of circulating antigen per ml of serum. Sera from 46 patients with paracoccidioidomycosis or other mycoses and sera from healthy individuals were evaluated by the inh-ELISA; overall sensitivity was 80.4% (37 of 46 paracoccidioidomycosis patients tested positive), and specificity compared with that of normal controls from areas of endemicity was 81.4%. The inh-ELISA detected circulating antigen in 100% of patients with the acute form of paracoccidioidomycosis and in 83.3 and 60% of patients with the chronic multifocal and unifocal forms of paracoccidioidomycosis according to the patients' clinical presentation. These results indicate that the inh-ELISA with MAb PIB is effective in the detection of circulating antigen and that this test may be useful for monitoring responses to treatment and establishing disease prognoses. PMID- 9399535 TI - Detection of antibodies to Candida albicans germ tubes for diagnosis and therapeutic monitoring of invasive candidiasis in patients with hematologic malignancies. AB - We prospectively investigated the ability of detection of antibodies to Candida albicans germ tubes (CAGT) to diagnose invasive candidiasis in 95 consecutive admissions of 73 patients with hematologic disorders undergoing intensive chemotherapy. The episodes were divided into three groups according to clinical and microbiological diagnosis. Group 1 comprised eight admissions of eight patients with invasive candidiasis. Group 2 comprised 42 admissions of 34 patients without evidence of invasive candidiasis. Group 3 comprised the remaining 45 admissions of 37 patients with febrile episodes which were not diagnosed by microbiological culture. Antibodies to CAGT were detected in 87.5% of group 1 patients. Detection of antibodies to CAGT in patients with Candida fungemia was delayed somewhat relative to the time the blood culture was positive, but antibodies to CAGT were detected earlier than a diagnosis was made in patients with deep-tissue candidiasis. Sera from 2 admissions in group 2 and 12 admissions in group 3 revealed antibodies to CAGT. At a titer of > or = 1:20, detection of antibodies to CAGT had a sensitivity of 87.5%, specificity of 95.2%, positive predictive value of 77.8%, and negative predictive value of 97.6%. Antibodies to CAGT were usually detected before beginning of empiric antifungal therapy. Titers of antibodies to CAGT were maintained in most patients who died but declined and eventually disappeared in the patients who survived. Since antibodies to CAGT were detected in all patients with tissue-proven invasive candidiasis but negative by blood culture, detection of antibodies to CAGT complemented blood cultures for diagnosis and therapeutic monitoring of patients with hematologic malignancies and invasive candidiasis. PMID- 9399536 TI - Comparison of the Amplicor HIV-1 monitor test and the nucleic acid sequence-based amplification assay for quantitation of human immunodeficiency virus RNA in plasma, serum, and plasma subjected to freeze-thaw cycles. AB - The Amplicor HIV-1 Monitor test was compared to the nucleic acid sequence-based amplification (Nasba) assay system for the quantitation of human immunodeficiency virus (HIV) RNA in three different types of clinical samples: plasma, serum, and plasma subjected to freeze-and-thaw cycles. Each assay detected HIV RNA in the same 73 (90%) of 81 samples tested, and the quantitative results obtained with the two assays were significantly correlated. Both assays detected higher RNA levels in patients with CD4+ cell counts lower than 200 cells/mm3 than in patients with CD4+ cell counts higher than 200 cells/mm3. In addition, RNA levels in plasma higher than 5 logs predicted higher numbers of clinical events than did RNA levels in plasma lower than 5 logs. Quantitation of HIV RNA in paired plasma and serum samples showed lower HIV RNA content in serum than in the paired plasma sample, with mean differences between HIV RNA contents of plasma and serum of 0.54 and 0.28 log RNA copy/ml by the Nasba assay and the Amplicor HIV-1 Monitor assay, respectively. No significant loss of HIV RNA was detected with either assay in plasma samples subjected to multiple freeze-and-thaw cycles. These studies demonstrate that the Nasba and Amplicor assays perform similarly with plasma and serum samples. Further, the results indicate that freeze-and-thaw cycles do not result in significant loss of detectable HIV RNA. PMID- 9399537 TI - Quantitative urine cultures do not reliably detect renal candidiasis in rabbits. AB - The significance of quantitative urine cultures in patients at risk for hematogenous disseminated candidiasis is controversial. While various concentrations of Candida spp. in urine have been suggested as critical cutoff points in the diagnosis of renal candidiasis, other investigators consider quantitative cultures less critical in diagnosing upper tract infections. To determine the significance of quantitative urine cultures in renal candidiasis, we studied serial quantitative urinary cultures of Candida albicans in a rabbit model of hematogenous infection. Of 197 urine samples from 34 infected animals, 144 were culture positive, with a sensitivity of 73.1% for urine cultures and a lower limit of detection of 10 CFU/ml. The yield of urine cultures varied according to severity and duration of infection. The mean renal and urinary concentrations of C. albicans from rabbits with subacute candidiasis differed significantly from those from rabbits with acute candidiasis (P = 0.013 and P < or = 0.001, respectively). During the first 4 days of subacute renal candidiasis, more than one-half of all urine cultures were negative for C. albicans. Only 12 (8.1%) of 148 urine cultures in animals with subacute renal candidiasis had concentrations of > 10(3) CFU/ml, 2.7% had concentrations of > 10(4) CFU/ml, and none were > or = 10(5) CFU/ml. By comparison, all urine cultures from the animals with lethal acute renal candidiasis had higher concentrations of C. albicans and were positive throughout the course of infection. Urinary concentrations of C. albicans were not predictive of the amount of Candida in the kidney (r < or = 0.49) and did not correlate with survival (r = 0.0232). However, the renal concentration of C. albicans (in CFU/gram) inversely correlated with the duration of survival (in days) of rabbits with renal candidiasis (r = 0.76; P < 0.001). These findings indicate that a negative urine culture in rabbits does not preclude the presence of renal candidiasis. The interpretation of a urine culture positive at any concentration, on the other hand, must involve an analysis of the risk factors for renal candidiasis, for any urinary concentration of C. albicans may reflect kidney infection. PMID- 9399538 TI - GEMHEP multicenter quality control study of PCR detection of GB virus C/hepatitis G virus RNA in serum. AB - PCR is, to date, the only available tool for the detection of GB virus C (GBV-C) and hepatitis G virus (HGV) RNAs. Twenty-two French laboratories participated in a quality control study to assess the sensitivity and specificity of their procedures. The panel included 13 positive controls and 7 negative controls. The laboratories used either in-house PCR techniques adapted from the literature or partly standardized commercial tests. Three laboratories performed faultlessly with the entire panel. Most laboratories had excellent specificity (100% in 20 of 22 laboratories). Sensitivity was acceptable (85 to 100%) in 15 centers and insufficient (38 to 77%) in 7. As with nonstandardized in-house PCR, the commercial assays gave discrepant performances in different laboratories. These results suggest that laboratories willing to use PCR for detection of GBV-C/HGV RNA for research or diagnostic purposes should participate in multicenter quality control trials. PMID- 9399540 TI - Surveillance of cytomegalovirus after solid-organ transplantation: comparison of pp65 antigenemia assay with a quantitative DNA hybridization assay. AB - In a multicenter study, 113 blood samples from 19 organ transplant patients were analyzed for cytomegalovirus by the pp65 antigenemia assay and a quantitative DNA hybridization assay. Overall, there was 84% agreement among the results obtained by the two tests. Fifteen of 16 episodes of active infection were detected by both assays. One episode was missed by the pp65 assay, and one patient showed significant DNA-emia but only low-level antigenemia. PMID- 9399539 TI - Reproducibility of AMPLICOR enterovirus PCR test results. AB - The reproducibility of AMPLICOR enterovirus PCR test results was determined with clinical samples of cerebrospinal fluid, serum, urine, and throat and rectal swabs. Among 608 samples from which duplicate aliquots were run simultaneously, only seven pairs gave discordant results. Among 104 samples from which duplicate aliquots were run in separate assays, no discordance was seen. Overall, the reproducibility of test kit results was 99% (705 of 712). PMID- 9399541 TI - Algaemia due to Prototheca wickerhamii in a patient with myasthenia gravis. AB - Prototheca wickerhamii is a rare cause of systemic infection in humans. While some cases occur in previously healthy individuals, others are associated with a variety of preexisting diseases. Here we present, for the first time, a case of P. wickerhamii algaemia in a patient with myasthenia gravis. The patient was successfully treated with amphotericin B. PMID- 9399542 TI - Comparison of the InPouch TV culture system and Diamond's modified medium for detection of Trichomonas vaginalis. AB - This study compared the use of Diamond's modified medium to InPouch for the culture of Trichomonas vaginalis from pooled vaginal secretions. The sensitivity for InPouch was 82.4% (61/74) versus 87.8% (65/74) for Diamond's modified medium. There were no significant differences in the sensitivity and negative predictive value of InPouch compared to Diamond's modified medium. PMID- 9399543 TI - Comparison of different methods for extraction of DNA of fungal pathogens from cultures and blood. AB - Five commercially available extraction kits and an in-house DNA extraction method for the release of DNA from Candida albicans and Aspergillus niger cells were assessed for sensitivity, purity, duration, and cost. All commercially available kits helped to shorten the duration of DNA extraction. However, the sensitivity varied from 1 to 1,000 fungal cells/ml and costs varied from $0.10 to 2.30. The QIAmp Tissue kit was the commercially available assay that yielded the same sensitivity and purity of fungal DNA release as the in-house protocol but was the most expensive method. In comparing these two extraction protocols, a 99% concordance of PCR results for 125 blood samples analyzed could be demonstrated. PMID- 9399544 TI - Detection of immunoglobulin G (IgG) and IgM antibodies to Toxoplasma gondii: evaluation of four commercial immunoassay systems. AB - A comparative evaluation of the following commercial immunoassays for the determination of antibodies to Toxoplasma gondii was performed: Behring Diagnostics OPUS Toxo G and Toxo M, Abbott Diagnostics IMX Toxo-IgG 2.0 and Toxo IgM, Sanofi Diagnostics Pasteur Platelia Toxo IgG and Toxo IgM, and bioMerieux Vitek VIDAS Toxo IgG and IgM. Of 676 specimens that were tested for Toxoplasma specific immunoglobulin G (IgG) antibodies, 26% were reactive by all methods while 8% displayed some discrepancy. Of 718 specimens that were tested for Toxoplasma-specific IgM antibodies, 3% were reactive by all methods while 10% displayed some discrepancy. Analysis of discrepant specimens revealed performance shortcomings with all IgM-specific assays. The impact of such shortcomings is magnified in a population with a low prevalence of toxoplasmosis. PMID- 9399545 TI - Serodiagnosis of American trypanosomosis by using nonpathogenic trypanosomatid antigen. AB - Crithidia luciliae, a nonpathogenic trypanosomatid, could provide a good alternative source of antigen for serodiagnosis of Chagas' disease. An enzyme linked immunosorbent assay showed 100% sensitivity and 83% specificity when 91 human serum samples from Chagas' disease patients and 127 human serum samples from people suffering from toxoplasmosis (21 samples), leishmaniasis (32 samples), systemic rheumatic diseases (33 samples), and heart diseases (41 samples) were tested simultaneously with Trypanosoma cruzi and C. luciliae crude extracts. By Western blotting, an immunodominant band (30 kDa) was recognized by chagasic sera on the C. luciliae crude extract; specificity reached 97% with respect to this protein band. The carbohydrate moieties were not antigenic. PMID- 9399546 TI - Increased levels of macrophage colony-stimulating factor, gamma interferon, and tumor necrosis factor alpha in sera of patients with Orientia tsutsugamushi infection. AB - Levels of macrophage colony-stimulating factor (nine of nine patients) and gamma interferon (six of nine patients) in serum were elevated above the range of normal in the acute phase of tsutsugamushi disease. Significant increases in levels of tumor necrosis factor alpha were observed during the convalescent phase in five patients, and they exceeded the levels observed during the acute phase. Hypercytokinemia appeared to be responsible for the emergence of the symptoms of tsutsugamushi disease. PMID- 9399547 TI - Susceptibilities of Chryseobacterium indologenes and Chryseobacterium meningosepticum to cefepime and cefpirome. AB - In vitro activities of cefepime and cefpirome against 96 isolates of Chryseobacterium indologenes and 21 of C. meningosepticum were determined by the agar dilution method. Overall, cefepime was more active than cefpirome against C. indologenes (MIC at which 50% of the isolates were inhibited [MIC50] and MIC90, 4 and 16 microg/ml, respectively, for cefepime and 8 and 128 microg/ml, respectively, for cefpirome). Both agents had poor potency against C. meningosepticum (MIC50 and MIC90, 64 and >256 microg/ml, respectively, for cefepime and 128 and >256 microg/ml, respectively, for cefpirome). PMID- 9399549 TI - Identification by spoligotyping of a caprine genotype in Mycobacterium bovis strains causing human tuberculosis. AB - We have used spoligotyping to characterize 18 Mycobacterium bovis strains isolated from cattle and 23 M. bovis strains isolated from goats. The spoligotypes revealed that caprine strains form a separate and well differentiated group that we refer to hereafter in this abstract as the caprine genotype. To evaluate the importance of this genotype as a cause of tuberculosis in other animal species, including humans, we applied the spoligotyping method to 112 strains, including to all isolates identified as M. bovis by a Mycobacterial National Reference Laboratory (Majadahonda, Madrid) from 1994 to 1996. Eighty three of these strains were identified in human isolates. In addition to being identified in three goat isolates and two sheep isolates, the caprine genotype was also found in three isolates causing human tuberculosis. Evidence to support the argument that there is a zoonotic risk of caprine tuberculosis was presented by the identification of the caprine genotype in an isolate from a veterinary worker with a recent history of contact with tuberculous goats. PMID- 9399550 TI - Rapid discrimination of Mycobacterium tuberculosis complex strains by ligation mediated PCR fingerprint analysis. AB - A ligation-mediated PCR (LMPCR) method for the amplification of sequences flanking the IS6110 of the Mycobacterium tuberculosis complex has been developed. The method uses one primer specific for IS6110 and a second specific for a linker ligated to SalI-restricted genomic DNA. LMPCR is a rapid screening method, valuable for the fingerprinting of M. tuberculosis complex strains. PMID- 9399548 TI - Reliability of mycobacteria growth indicator tube for testing susceptibility of Mycobacterium tuberculosis to ethambutol and streptomycin. AB - The reliability of mycobacterial growth indicator tubes (MGIT) for testing susceptibility of Mycobacterium tuberculosis to ethambutol and streptomycin was evaluated by comparing MGIT results to those obtained by the radiometric BACTEC TB system and the method of proportion. The method of proportion was considered the reference method. To resolve discrepancies, all three testing methods were repeated. For the 74 isolates evaluated, initial ethambutol results agreed by all three methods for 64 (86.5%) of them; 58 were susceptible and 6 were resistant. MGIT and method-of-proportion results agreed for 67 isolates, and BACTEC results agreed with method-of-proportion results for 71 (P = 0.096). Initial streptomycin results obtained by all three methods agreed for 69 (93.2%) isolates: 55 were susceptible and 14 were resistant. MGIT and method-of-proportion results were concordant for 69 isolates, and BACTEC and method-of-proportion results agreed for 73 (P = 0.049). The mean times to MGIT results were 5.58 +/- 0.10 days (range, 3 to 9 days) for ethambutol and 5.47 +/- 0.11 days (range, 3 to 9 days) for streptomycin, compared to a mean of 7.41 +/- 0.20 days (range, 4 to 12 days) for both drugs with the BACTEC system (P < 0.001). PMID- 9399552 TI - Peritonitis associated with a CDC group EO-3 organism. AB - A 63-year-old female with chronic renal failure on continuous ambulatory peritoneal dialysis developed chronic peritonitis. A CDC group EO-3 organism was isolated from the peritoneal dialysis fluid on five occasions over a period of 4 months. This is the first reported isolation of this organism in which it is associated with a patient on continuous ambulatory peritoneal dialysis. PMID- 9399551 TI - Necrotizing pneumonia caused by Penicillium chrysogenum. AB - We report a case of necrotizing pneumonia due to Penicillium chrysogenum in a 57 year-old woman operated on for lung cancer. The residual right lower pulmonary lobe was infiltrated by Penicillium chrysogenum. The patient underwent a second pulmonary right lobectomy and was successfully treated with oral itraconazole. To our knowledge, this is the first case of pneumonia due to P. chrysogenum. PMID- 9399553 TI - Isolation of Cryptococcus neoformans var. gattii from Eucalyptus camaldulensis in India. AB - Cryptococcus neoformans var. gattii has an ecological association with five Eucalyptus species: E. blakelyi, E. camaldulensis, E. gomphocephala, E. rudis, and E. tereticornis. After human infections due to C. neoformans var. gattii were diagnosed in the states of Punjab, Himachal Pradesh, and Karnataka, India, a study was undertaken to investigate the association of C. neoformans var. gattii with Indian eucalypts, especially in the state of Punjab. A total of 696 specimens collected from E. camaldulensis, E. citriodora and E. tereticornis (hybrid) trees were examined for the presence of C. neoformans var. gattii. Flowers from two trees of E. camaldulensis in the Chak Sarkar forest and one from the village of Periana near the Ferozepur area yielded five isolates of C. neoformans var. gattii. The origin of the trees could be traced to Australia, thus providing evidence that the distribution of E. camaldulensis correlated with the distribution of human cryptococcosis cases caused by C. neoformans var. gattii in northern India. PMID- 9399554 TI - Cutaneous hyalohyphomycosis caused by Fusarium solani in a loggerhead sea turtle (Caretta caretta L.). AB - Fusarium solani was reported as the agent of a cutaneous infection in an injured sea turtle collected in the Mediterranean Sea. The turtle was treated with both a topical 10% solution of iodine in alcohol and ketoconazole. The source of the causal agent was traced to the sand in the tank in which the turtle was maintained. The strain was only sensitive in vitro to amphotericin B and was resistant to 5-fluorocytosine, fluconazole, itraconazole, and ketoconazole. PMID- 9399555 TI - Yersinia enterocolitica serotype O:8 septicemia in an otherwise healthy adult: analysis of chromosome DNA pattern by pulsed-field gel electrophoresis. AB - We report the first case of blood culture-positive Yersinia enterocolitica serotype O:8 septicemia in Japan. Y. enterocolitica serotype O:8 infection is very rare, but chromosomal DNA analysis suggested that this bacterium may persist latently in healthy carriers throughout Japan. PMID- 9399556 TI - Unique organization of the CTX genetic element in Vibrio cholerae O139 strains which reemerged in Calcutta, India, in September 1996. AB - We studied the restriction fragment length polymorphism of the rRNA gene and CTX genetic element in Vibrio cholerae O139 Bengal, which resurged in Calcutta in September 1996 after a gap of 32 months. While the strains from this resurgence were indistinguishable from the earlier strains by ribotyping, the structure of the CTX genetic element present in the current O139 strains was found to be unconventional. PMID- 9399557 TI - Healthy puppies and kittens as carriers of Campylobacter spp., with special reference to Campylobacter upsaliensis. AB - Living in a household with a dog or cat has previously been identified as a significant risk factor for acquiring campylobacteriosis, in particular, with reference to Campylobacter upsaliensis infection. In a cross-sectional study carried out in Denmark between August and December 1996, 72 healthy puppies and 42 healthy kittens, aged between 11 and 17 weeks, were sampled for fecal campylobacter shedding by culture of rectal swab specimens on blood-free agar base with cefoperazone at 32 mg/liter and amphotericin at 10 mg/liter and on blood-free agar base with cefoperazone at 8 mg/liter, teicoplanin at 4 mg/liter, and amphotericin at 10 mg/liter. Additionally, with respect to the C. upsaliensis transmission potential of poultry, a chicken cloacal swab sample from each of 100 different broiler flocks was included in the study for comparison. We found 21 (29%) of the puppies positive for Campylobacter spp., with a species distribution of 76% C. jejuni, 5% C. coli, and 19% C. upsaliensis. Of the kittens examined, two (5%) excreted campylobacters; both strains were C. upsaliensis. None of the chicken samples examined were found to be positive for C. upsaliensis. We concluded that young puppies and kittens are potential transmitters of human pathogenic Campylobacter spp., including C. upsaliensis, while poultry seems negligible in C. upsaliensis epidemiology. PMID- 9399558 TI - Citrobacter farmeri bacteremia in a child with short-bowel syndrome. AB - A case of sepsis in a pediatric patient due to the newly described Citrobacter species C. farmeri is described. Factors predisposing this child to infection included short-bowel syndrome requiring total parenteral nutrition. PMID- 9399559 TI - Detection of Chlamydia trachomatis in urine samples by nucleic acid tests: comparison with culture and enzyme immunoassay of genital swab specimens. AB - Two commercially available nucleic acid-based tests, ligase chain reaction (LCR; Abbott Laboratories) and PCR (Roche Diagnostics), for the detection of Chlamydia trachomatis in male and female urine samples were compared with culture and enzyme immunoassay (EIA) (Microtrak; Syva) for C. trachomatis detection in genital samples. The samples were collected from 1,005 patients who attended a sexually transmitted disease clinic. In this study population, the prevalence of the infection was 4%. Specimens which were reactive in any of the tests were retested with a different PCR test using primers directed against the major outer membrane protein gene. With a "gold standard" of a positive culture, or any other positive test result if it was confirmed by an independent test, the Roche PCR (95% sensitive, 99.9% specific) was more sensitive than the LCR (75% sensitive, 100% specific) (chi2, P < 0.0001) while both tests were more sensitive than culture (58% sensitive, 100% specific) or EIA (45% sensitive, 100% specific) (chi2, P < 0.001). The Roche PCR and Abbott LCR tests of urine identified 65% and 30% more positive patients, respectively, than did testing by culture of urethral or cervical specimens. Nucleic acid testing of urine specimens for C. trachomatis is a more sensitive and convenient method for the detection of genital infection. PMID- 9399560 TI - Salivary immunoglobulin G assay to diagnose Helicobacter pylori infection in children. AB - An in-house enzyme-linked immunosorbent assay (ELISA) for measurement of Helicobacter pylori-specific immunoglobulin G (IgG) and IgA in saliva was evaluated by comparison with histopathologic (Giemsa staining) and biochemical (urease quick test) examination of gastric biopsy specimens obtained from 112 children referred for diagnostic gastroscopy. Serum H. pylori IgG was also measured in a subgroup of 50 children by the same ELISA. Salivary H. pylori IgG levels were significantly higher in H. pylori-positive (n = 57) than in H. pylori negative (n = 55) children (P < 0.001). The sensitivity and specificity of the salivary IgG test were 93 and 82%, respectively; the positive and negative predictive values were 84 and 92%, respectively; and the accuracy was 87.5%. Salivary H. pylori IgA did not distinguish H. pylori-positive from H. pylori negative children. The performance of serum H. pylori IgG was slightly (3 to 6%) better than that of salivary H. pylori IgG. The salivary IgG test can be considered a useful tool for the screening of H. pylori infection in children. PMID- 9399561 TI - Reproducibility and performance of the AMPLICOR Chlamydia trachomatis test. PMID- 9399562 TI - Shewanella putrefaciens abscess of the lower extremity. PMID- 9399563 TI - PCR-based detection of Vibrio cholerae O139 Bengal with sequences encoding glycosyltransferases. PMID- 9399565 TI - Inhibitory properties of a swab transport device. PMID- 9399564 TI - Coexistence of Ehrlichia phagocytophila and Borrelia burgdorferi sensu lato in Ixodes ricinus ticks from Italy as determined by 16S rRNA gene sequencing. PMID- 9399566 TI - Mistyping of two Slovenian hepatitis C virus subtype 2c isolates as subtype 2b by two 5' noncoding region genotyping methods. PMID- 9399567 TI - Crystallization and preliminary X-ray analysis of a truncated family A alkaline endoglucanase isolated from Bacillus sp. KSM-635. AB - The catalytic domain of an alkaline endo-1,4-beta-glucanase (family A) isolated from Bacillus sp. KSM-635 (Mr = 40.2 kDa) was crystallized using the hanging drop vapor diffusion method. Two different crystal forms were obtained. Form 2 crystals (trigonal space group R3 with cell dimensions of a = b = 111.9 and c = 207.1 angstroms in a hexagonal lattice) were found to be more stable than form 1 ones upon X-ray irradiation. A full data set for form 2 crystals has been collected up to 3.3 angstroms resolution. PMID- 9399568 TI - Construction of transforming growth factor alpha (TGF-alpha) phage library and identification of high binders of epidermal growth factor receptor (EGFR) by phage display. AB - TGF-alpha, a 50 amino acid growth factor containing 3 disulfide bonds, was fused to the N-terminal domain of the pIII protein of fusN, a derivative of phagemid fd tet, to form a TGF-alpha phage. The fusion phage showed binding activity to epidermal growth factor receptor (EGFR). A library of approximately 4 x 10(7) variants of TGF-alpha was generated with substitutions of total of 10 amino acids located in the C-loop region. This C-loop subdomain of TGF-alpha consists of a small antiparallel double hairpin structure involving interactions between intra polypeptide segments. Mutants isolated from the phage library with greatly increased binding affinity were selected through panning with A431 cells (a cell line expressing an elevated number of EGFRs). Following two rounds of stringent selection, variant phages with higher binding affinity than wild type TGF-alpha were identified and the phage DNAs were sequenced for the alignment analysis. Absolute selection at position 42 as Arg, preferential selection at position 38 and 45 as Tyr or Phe with aromatic side chain and selection at position 41 with acidic residues, were obtained. Although an amino acid residue with smaller side chain at position 35 and one with larger side chain at position 36 were preferred, the steric hindering of the structure in side chains was minimized between these adjacent amino acids. PMID- 9399569 TI - Expression of the human UDP-galactose transporter in the Golgi membranes of murine Had-1 cells that lack the endogenous transporter. AB - In our previous study, we demonstrated that UDP-galactose transporter cDNAs (hUGT1 and hUGT2) were able to complement the genetic defect of murine Had-1 cells that were deficient in the UDP-galactose transporter, and that the microsomal vesicles isolated from Had-1-transformants, which were obtained through transfection with these cDNAs, had recovered the ability to uptake UDP galactose [Ishida, N. et al. (1996) J. Biochem. 120, 1074-1078]. In this report, we describe the preparation of peptide antibodies that recognize the hUGT isozymes, and the detection of hUGT proteins expressed in the transformants. The occurrence of the endogenous hUGT1 protein in HeLa cells was also detected. Using the hUGT1-specific antibodies, the subcellular localization of hUGT1 in the Golgi membrane was demonstrated by immunofluorescence microscopy and subcellular fractionation. These studies led us to develop a simple procedure, based on Percoll density gradient centrifugation, for preparing functional Golgi vesicles from the hUGT1-transformed Had-1 cells, that will facilitate future biochemical analyses of the UDP-galactose transporter for the elucidation of its structure function relationship. PMID- 9399570 TI - Structure and function in Escherichia coli of plasmids containing pyrimidine/purine-biased stretch originated from the 5'-flanking region of the basidiomycete ras gene. AB - The Basidiomycete ras gene possesses a pyrimidine-rich stretch (CT-motif) with a short (7 bases) mirror repeat in which its major transcription start point is contained. To analyze the tertiary structure induced by the CT/AG-biased sequence and its effect on gene expression in supercoiled plasmids in Escherichia coli, the DNA fragment containing the ras CT/AG sequence was inserted into the EcoRI site on pBR322 in both orientations and the resulting pBR322 derivatives, named pBR-CT[ras] and pBR-invCT[ras] were introduced into E. coli strains DM800 (deltatopA gyrB225) and JM109 (topA+ gyrA96). In pBR-CT [ras] the pyrimidine-rich sequence is on the pBR322 tetracycline-resistance gene (tet)coding strand and in pBR-invCT[ras] the complementary purine-rich sequence is on this strand. DNAs of pBR-CT[ras] and pBR-invCT[ras] isolated from DM800 were frequently cleaved with single-strand-specific S1 nuclease within the CT/AG sequence, showing the formation of extended open structure. Compared with those carrying pBR322, DM800 and JM109 carrying pBR-CT [ras] showed much higher levels of tetracycline resistance (Tcr), while both strains carrying pBR-invCT[ras] showed clearly lower levels of Tcr. pBR-CT [ras] and pBR-invCT [ras], however, conferred reduced activity of beta-lactamase on DM800 and JM109. pBR-CT [ras] derivatives lacking the counterpart of the mirror repeat did not form the S1-cleavable open structure within the CT/AG sequence and conferred pBR322-like Tcr and beta-lactamase activity. The tertiary structure formed in the CT/AG sequence via the mirror repeat was suggested to affect the expressions of pBR322-tet and -bla genes. PMID- 9399571 TI - CBP/p300 functions as a possible transcriptional coactivator of Ah receptor nuclear translocator (Arnt). AB - A heterodimer of AhR (aryl hydrocarbon receptor) and Arnt (AhR nuclear translocator) conveys a transactivation signal of aromatic hydrocarbons such as 2,3,7,8-tetrachlorodibenzo-p-dioxin and 3-methylcholanthrene to the genes for a group of drug-metabolizing enzymes. This inducible expression of the genes is inhibited by adenovirus E1A, suggesting that CBP/p300 is somehow involved in the transactivation of the genes by the AhR and Arnt heterodimer. Yeast and mammalian two hybrid systems revealed that CBP/p300 interacted with the transactivation domain of Arnt, but not with that of AhR, via the CREB-binding domain. The pull down assay using GST-Arnt hybrid protein confirmed the interaction between Arnt and CBP/p300. Considering these results and that Arnt or Arnt2 functions as a common partner in the formation of transcriptional regulators with other bHLH/PAS proteins such as AhR, HLF, and HIF-1alpha, the possibility arises that CBP/p300 is extensively involved as a coactivator in the transactivation process by bHLH/PAS (a conserved sequence motif among Per, Arnt, and Sim) heterodimer transcription factors through interaction with Arnt or Arnt2. PMID- 9399572 TI - Dimerization and DNA binding facilitate alpha-helix formation of Max in solution. AB - Max is a basic region/helix-loop-helix/leucine zipper (b/HLH/Z) protein that forms a hetero-complex with the Myc family proteins Myc, Mad, and Mxi1, and a homo-complex with itself. These complexes specifically bind to double-stranded DNA containing CACGTG sequences. Here, we report on the structural properties in aqueous solution of a 109-amino-acid protein, Max110, corresponding to the N terminal domain of Max containing the b/HLH/Z motif (residues 2-110), as characterized by combined use of circular dichroism (CD) and sedimentation equilibrium experiments. The results showed that the alpha-helical content of Max110 increases with increasing protein concentration. The sedimentation equilibrium data indicated that Max110 exists as a monomer at low protein concentration, and forms a dimer at high protein concentration. Further increases in the alpha-helical content of Max110 occur upon addition of DNA with the CACGTG recognition sequence. Thus, dimerization and binding to DNA of Max both favor an increase of the alpha-helical content. PMID- 9399573 TI - Antisense oligodeoxynucleotides of IGF-II selectively inhibit growth of human hepatoma cells overproducing IGF-II. AB - Insulin-like growth factor II (IGF-II) is expressed in many developing embryonic tissues and is involved in mammalian growth and development. After birth, serum IGF-II is mainly produced by liver cells. Many reports have indicated that IGF-II is overexpressed in some hepatocellular carcinoma (HCC) tissue. These findings imply the possible importance of this growth factor in carcinogenesis. We screened four human HCC cell lines and three rat HCC cell lines and found that HuH-7 and HepG2 cells produced fivefold more intracellular IGF-II than the other cell lines. Experimental data indicate that IGF-II functions through the intracrine mode for HuH-7 cells. To study whether the overexpression of IGF-II is significant for the growth of HCC or only a consequence of HCC development, we used antisense oligodeoxynucleotides (ATON) to arrest the translation of IGF-II mRNA, and then measured the effects on cell growth. We found that the production of IGF-II was suppressed by ATON, and the decrease of IGF-II resulted in growth inhibition of HuH-7 and HepG2. ATON had no effect on the other tested cell lines, which produced lower levels of IGF-II. The growth inhibition was mainly attributed to a decrease of cell proliferative activity. The results indicate that the IGF-II-overproducing cell lines do depend on IGF-II for growth, and ATON of IGF-II can selectively inhibit the growth of these cells. ATON may be a potential therapeutic agent for this type of HCC in vivo. PMID- 9399574 TI - Relationships of subunits of type-1 serine/threonine protein phosphatase to morphology and aggregation of B cells. AB - To elucidate the roles of serine/threonine protein phosphatases PP1 and PP2A in the morphological changes of B-lymphocytes during development and in immune responses, we investigated alterations of protein levels of catalytic subunits of PP1 and PP2A and regulatory subunits of PP1 including M130/M133, inhibitor-1 (I 1) and inhibitor-2 (I-2) in B-cell lines at different maturational stages and during their aggregation induced by phorbol myristate acetate (PMA). The protein levels of PP1delta and/or M130/M133 were significantly lower in B-cell lines without pseudopods, WEHI-231, BAL-17, Daudi, and CESS, than in those with pseudopods, Bcl.1, A20, M12, and SKW6.4, whereas the amounts of PP1alpha and PP2A were similar among them. During aggregation of A20 and CESS cells induced by PMA, an activator of PKC, the amount of PP1delta was progressively decreased, and this decrease was blocked by H7, an inhibitor of PKC. The amount of PP1alpha was constant under these conditions. Okadaic acid, an inhibitor of PP1 and PP2A, also induced aggregation of A20 cells at concentrations sufficient to inhibit PP1, but not at lower concentrations that inhibit PP2A alone. These results suggest that myosin light chain phosphatase composed of PP1delta and M130/M133 is involved in the maintenance and regulation of cytoskeletal structures in B-lymphocytes. PMID- 9399575 TI - Alterations in type-1 serine/threonine protein phosphatase PP1alpha in response to B-cell receptor stimulation. AB - In response to stimulation of B-cells through cell surface IgM, the activity of the serine/threonine protein phosphatase PP1, but not PP2A, was transiently decreased and reached a minimum 10-20 min after the stimulation. The decrease was more profound in the immature B-cell line WEHI-231, than in the mature B-cell line BAL-17. Under these conditions, PP1alpha, an isoform of PP1, showed unique alterations in the patterns of several spots with distinct isoelectic points in the Western blot after two-dimensional electrophoresis, whereas another isoform, PP1delta, did not show any alteration. PP1gamma1 and PP1gamma2 were not detected in B-cells. Similar alterations in these spots were observed in B-cells stimulated by PMA. When partially purified PP1 consisting of PP1alpha and PP1delta was incubated with [gamma-32P]ATP and PKC, radioactive spots of PP1alpha could be detected, but no spot of PP1delta was detected. Because differences in sequence among PP1 isoforms are mostly restricted to their C-terminals, phosphorylation rates of the C-terminal peptides containing the PKC phosphorylation motif were compared. The C-terminal peptide of PP1alpha is a better substrate for PKC than those of PP1gamma1 and PP1gamma2, and is phosphorylated at the serine residue corresponding to Ser-325 of PP1alpha. The corresponding C-terminal region of PP1delta does not contain the phosphorylation site. On the other hand, there was a large difference in subcellular distribution of PP1delta, but not PP1alpha, between immature and mature B-cells. From these results, it was strongly suggested that PP1alpha is involved, via phosphorylation by PKC, in the regulation of signal transduction in response to the stimulation of B-cells through cell surface IgM. PMID- 9399576 TI - Inhibition of phosphoinositide hydrolysis and cell growth of Swiss 3T3 cells by myristoylated phospholipase C inhibitor peptides. AB - It has been demonstrated that the phospholipase C-gamma (PLC-gamma) molecule contains within it a phospholipase C inhibitor (PCI) region and that synthetic peptides based on the sequence of this region (PCI peptides) suppress the enzymatic activity of PLC isoforms [Y. Homma and T. Takenawa (1992) J. Biol. Chem. 267, 21884-21889]. In order to improve the permeability of the plasma membrane to PCI peptides, we synthesized myristoylated PCI peptides, myr GLYRKAMRLRYPV [myr-PCI(Y)] and myr-GLFRKAMRLRFPV [myr-PCI(F)], which are identical except for the replacement of the two tyrosine residues in myr-PCI(Y) by phenylalanines in myr-PCI(F), and examined their inhibitory activity on PLC enzymes in vitro and in vivo. This fatty acid modification potentiated the inhibitory activity of the original PCI peptides and both myr-PCI(Y) and myr PCI(F) suppressed the PIP2-hydrolyzing activity of purified PLC isoforms in vitro. The Ki values of myr-PCI(Y) and myr-PCI(F) for purified PLC-gamma1 were 3.5 and 55 microM, respectively. Myr-PCI(Y) at concentrations in the sub micromolar range significantly suppressed IP3 formation induced by EGF, PDGF, bombesin, or serum in Swiss 3T3 cells. Furthermore, myr-PCI(Y) also strongly inhibited cell proliferation induced by these stimuli. The inhibitory effect on IP3 formation and proliferation of myr-PCI(F) was much less potent than that of myr-PCI(Y). These results suggest that myristoylated PCI peptides could be applied to living cells as specific inhibitors of PLC signaling pathways and that PLC pathways are at least in part required for growth in Swiss 3T3 cells. PMID- 9399577 TI - Downregulation of calpastatin in rat heart after brief ischemia and reperfusion. AB - The activities of calpain and its endogenous inhibitor, calpastatin, were measured in the soluble fraction of perfused rat heart after ischemia for 5-20 min and reperfusion for up to 30 min. The method for m-calpain measurement was modified: washing of the DEAE-cellulose column with 0.18 M NaCl instead of 0.15 M NaCl increased the m-calpain activity 12.5-fold. Ischemia for 20 min followed by reperfusion for 30 min did not affect the m-calpain activity but decreased the calpastatin activity. m-Calpain was enriched in the nucleus-myofibril fraction but was not further translocated on ischemia-reperfusion. Mu-calpain was below the limit of detection on immunoblotting or casein zymography, but its mRNA was substantially expressed, as detected on Northern blotting. Casein zymography also revealed a novel Ca2+-dependent protease without the typical characteristics of mu- or m-calpain. The immunoblotting of myocardial fractions showed that calpastatin was proteolyzed on ischemia-reperfusion. The calpastatin proteolysis was suppressed by a calpain inhibitor, Ac-Leu-Leu-norleucinal. Calpastatin may sequester calpain from its substrates in the normal myocardium, but may be proteolyzed by calpain in the presence of an unidentified activator in the early phase of calpain activation during ischemia-reperfusion, resulting in the proteolysis of calpastatin and then other calpain substrates. PMID- 9399578 TI - Cleavage activity of hepatitis C virus serine proteinase. AB - To study the character of the hepatitis C virus (HCV) encoding serine proteinase and to search for inhibitors, a practical in vitro assay system using the purified enzyme and synthetic peptide substrates was established. The enzyme used was expressed in Escherichia coli as a fusion form with protein tags and purified to apparent homogeneity by single-step affinity chromatography. The purified enzyme exhibited proteolytic activity with pH optima of around eight, and the addition of NS4A fragments increased the activity as well as the thermal stability of the enzyme. The activity was inhibited by EDTA and some divalent ions, i.e., copper and zinc, though calcium, magnesium, and manganese were stimulative both in the presence and absence of the NS4A fragment. None of the common protease inhibitors, including serine protease inhibitors, effectively inhibited the activity. Based on the kinetic parameters of the cleavage reaction of the synthetic 20 mer peptides corresponding to the three cleavage sites, NS4A/4B, NS4B/5A, and NS5A/5B, the peptide with the NS5A/5B junction was found to be the most efficient substrate. Analysis of the minimal peptide substrate of NS5A/5B indicated that 5 to 7 amino acids on both sides of the junction were required for efficient cleavage. These findings are expected to contribute to the search for a proteinase inhibitor. PMID- 9399579 TI - Identification and characterization of a major lysosomal membrane glycoprotein, LGP85/LIMP II in mouse liver. AB - We previously have purified and characterized a major lysosomal membrane glycoprotein termed LGP85 (LIMP II) in rat liver lysosomes. In this study, LGP85 in mouse liver lysosomes was identified and characterized by biochemical and molecular biological methods. Lysosomal membranes were isolated from murine liver by differential centrifugation. LGP85 was present in the lysosomal membrane fraction from mouse liver in a comparable amount to another lysosomal membrane glycoprotein, lamp-2. Mouse LGP85 (M-LGP85) from liver lysosomal membranes exhibited an Mr of 80,000 on SDS-PAGE, which is smaller by 5,000 than that of rat LGP85 (R-LGP85). M-LGP85 was immunochemically detected in the extracts of brain, heart, lung, liver, and kidney. A cDNA encoding M-LGP85 was cloned from mouse liver cDNA library. The primary protein structure deduced from a nucleotide sequence of M-LGP85 cDNA indicated that M-LGP85 consists of 478 amino acids with Mr of 54,069. M-LGP85 showed 93.3 and 86.0% sequence similarities to its rat and human counterparts in amino acids, respectively. M-LGP85 contains 11 potential N glycosylation sites which are heavily glycosylated, resulting in the increased Mr of M-LGP85 present in the mouse liver lysosomes. It is likely that M-LGP85 traverses the lysosomal membrane twice, with an NH2-terminal transmembrane domain, and another hydrophobic domain near the COOH-terminus. M-LGP85 has a protruding COOH-terminal cytoplasmic tail consisting of amino acid residues including the leucine-isoleucine sequence shown to be the lysosomal targeting signal of R-LGP85 and human LGP85 (H-LGP85). The high level of expression of M LGP85 in the lysosomal membrane, the high structural similarities among M-, R-, and H-LGP85, and the occurrence of M-LGP85 in all the mouse tissues examined suggest the essential and constitutive function of LGP85 in lysosomes. PMID- 9399580 TI - A novel cytochrome b(o/a)3-type oxidase from Bacillus stearothermophilus catalyzes cytochrome c-551 oxidation. AB - Gram-positive thermophilic Bacillus species contain cytochrome caa3-type cytochrome c oxidase as their main terminal oxidase in the respiratory chain. To identify alternative oxidases, we isolated several mutants from B. stearothermophilus defective in the caa3-type oxidase activity [Sakamoto, J. et al (1996) FEMS Microbiol. Lett. 143, 151-158]. A novel oxidase was isolated from membrane preparations of one of the mutants, K17. The oxidase was composed of two subunits with molecular masses of 56 and 19 kDa, and contained protoheme IX, heme O, heme A, and Cu in a ratio of 1:0.7:0.2:3. CO difference spectra indicate that the high-spin heme is mainly heme O. These results suggest that the enzyme belongs to the heme-copper oxidase family and is a cytochrome b(o/a)3-type oxidase, whose high-spin heme is mainly heme O and partly heme A. The enzyme oxidized cytochrome c-551, which is a membrane-bound lipoprotein of thermophilic Bacillus. The turnover rate of the activity (Vmax = 190 s[-1]) and its affinity for cytochrome c-551 (Km = 0.15 microM) were much higher than those for yeast and equine heart cytochromes c. The oxidase activity was enhanced by the presence of salts and inhibited by sodium cyanide with a Ki value of 19 microM. The enzyme kinetics suggests that cytochrome c-551 is the natural substrate to this oxidase. Furthermore, the oxidase had similarity to cytochrome ba3-type oxidase from Thermus thermophilus in the subunit composition, partial amino acid sequence, and prosthetic groups, and therefore is suggested to belong to a unique subgroup of the heme-copper oxidase family together with the Thermus enzyme and archaeal oxidases such as Sulfolobus SoxABCD. PMID- 9399581 TI - Purification, staphylolytic activity, and cleavage sites of alpha-lytic protease from Achromobacter lyticus. AB - Alpha-lytic protease (alp) was purified from a bacteriolytic agent, Achromopeptidase from Achromobacter lyticus M497-1, and has been shown to possess staphylolytic activity. Cleavage sites of this enzyme on the peptidoglycan of Staphylococcus aureus were determined by N-terminal amino acid sequence and amino acid composition analyses. Alp cleaved the N-acetylmuramoyl-L-alanine amide bond, the junction between the polysaccharide and peptide moieties, in addition to the D-Ala-Gly and Gly-Gly peptide bonds, implying that this enzyme recognizes the amino acid of D-configuration at the P1 site and possesses N-acetylmuramoyl-L alanine amidase activity. However, alp could not cleave the D-Ala-Gly peptide bond in a synthetic peptide, suggesting that this hydrolytic activity of alp is peptidoglycan-specific. The results obtained from different consecutive actions of alp and glycosidase on S. aureus peptidoglycan indicate that the presence of polysaccharide in the peptidoglycan is necessary for the bacteriolytic activity of alp. PMID- 9399582 TI - Alanyl aminopeptidase from human seminal plasma: purification, characterization, and immunohistochemical localization in the male genital tract. AB - Alanyl aminopeptidase (AAP) was purified to homogeneity from human seminal plasma. The calculated molecular weight of the purified enzyme was approximately 137,000+/-5,000 from light scattering, 140,000 (main) and 137,000 (minor) from non-denatured PAGE and 153,000 from SDS-PAGE in the absence or presence of 2 mercaptoethanol (2-ME). These findings suggest that the enzyme is monomeric in form in human seminal plasma. The enzyme hydrolyzed several amino acid 4-methyl coumaryl-7-amide (MCA) substrates. The order of Kcat/Km values of AAP at optimal pH (pH 7.5) was Ala- > Lys-Ala- > or = Met- > Leu- > Phe- > Arg- > or = Arg-Arg- > Lys- > Gly-MCAs. AAP was potently inhibited by bestatin, leuhistin, actinonin, amastatin, and 1,10-phenanthroline. These findings suggest that AAP is an aminopeptidase. We determined that the amino acid sequence of the first 22 residues of the enzyme was Ser1-Thr-Thr-Pro-Ser5-Ala-Ser-Ala-Thr-Thr10-Asn-Pro-Al a-Ser-Ala15-Thr-Thr-Leu-Asp-Gln20-Ser-Lys-. This sequence was completely coincident with that downstream of the transmembrane site of human intestinal alanyl aminopeptidase N (CD13). We also isolated cDNA encoding AAP from human prostate cDNA library, sequenced its structure, and confirmed human seminal plasma AAP to be identical with alanyl aminopeptidase N. We postulated that native human seminal plasma alanyl aminopeptidase is released into the seminal plasma after the specific site is cleaved by elastase or an elastase-like enzyme. The enzyme level in human seminal plasma determined by single radial immunodiffusion was 5.2+/-3.2 mg/100 ml (mean+/-SD, n=20) in individuals 20-47 years of age. AAP was immunohistochemically stained in the luminal site-cell membrane of epithelial cells in the prostatic gland and ductuli efferentes of the testis. PMID- 9399583 TI - Resynthesis of reactive site peptide bond and temporary inhibition of Streptomyces metalloproteinase inhibitor. AB - Streptomyces metalloproteinase inhibitor (SMPI) is a small proteinaceous inhibitor which inhibits metalloproteinases such as thermolysin (Ki =1.14 x 10( 10) M). When incubated with the enzyme, it is gradually hydrolyzed at the Cys64 Val65 peptide bond, which was identified as the reactive site by mutational analysis. To achieve a further understanding of the inhibition mechanism, we attempted to resynthesize the cleaved reactive site by using the enzyme catalytic action. The native inhibitor was resynthesized from the modified inhibitor (Ki =2.18 x 10(-8) M) by incubation with a catalytic amount of thermolysin under the same conditions as used for hydrolysis (pH 7.5, 25 degrees C), suggesting that SMPI follows the standard mechanism of inhibition of serine proteinase inhibitors. Temporary inhibition was observed when the native inhibitor and thermolysin were incubated at a 1:100 (mol/mol) enzyme-inhibitor ratio at 37 degrees C. SMPI showed temporary inhibition towards all the enzymes it inhibited. The inhibitory spectrum of SMPI was analyzed with various metalloproteinases based on the Ki values and limited proteolysis patterns. Pseudomonas elastase and Streptomyces griseus metalloproteinase II formed more stable complexes and showed much lower Ki values (approximately 2 pM) than thermolysin. In the limited proteolysis experiments weak inhibitors were degraded by the enzymes. SMPI did not inhibit almelysin, Streptomyces caespitosus neutral proteinase or matrix metalloproteinases. SMPI specifically inhibits metalloproteinases which are sensitive to phosphoramidon. PMID- 9399584 TI - Genomic organization of the rat nuclear factor I-A gene. AB - The nuclear factor 1 (NF1) protein family functions as a cellular transcription factor as well as an adenovirus DNA replication factor. This family consists of four subtypes, NFI-A, NFI-B, NFI-C, and NFI-X, each encoded by a different gene. Each subtype possesses different isoforms generated by alternative splicing. To date, only a porcine NFI-C gene has been cloned, and the gene structures of the other NF1 proteins have not yet been identified. We recently isolated four kinds of NFI-A cDNA clones from the rat liver. To gain additional insight into the structure of NFI-A, we isolated the rat NFI-A gene. This gene is composed of 11 exons spanning over 70 kb. All of the exon/intron boundaries are consistent with the GT/AG rule, and consensus sequences surrounding the splice boundaries are also found. The 5'-flanking region lacks a canonical TATA box, but contains several GC-box and AP2 binding sites. A 5'-rapid amplification of cDNA end analysis indicated that the transcription of the NFI-A gene is initiated at multiple sites. We also found conservation in the genomic structure between the rat NFI-A and the porcine NFI-C, suggesting that duplication of an ancestral gene occurred rather recently to produce the NFI-A and NFI-C genes. PMID- 9399585 TI - Involvement of Glu-264 and Arg-235 in the essential interaction between the catalytic imidazole and substrate for the D-lactate dehydrogenase catalysis. AB - For Lactobacillus pentosus D-lactate dehydrogenase, the binding of 2-ketoacids is markedly stabilized through interactions between the protonated imidazole of His 296, an acid/base catalyst of the enzyme, and the carbonyl oxygen of 2-ketoacids. The replacement of Arg-235 with Gln destabilized the inhibitory binding of oxamate much more than that of formate, acetate, or propionate, and the Arg to Lys substitution specifically diminished only oxamate binding. On the other hand, replacement of a conserved Glu, Glu-264, with Gln severely impaired the enzyme activity and markedly reduced affinity to 2-keto acids. The pH dependence of the oxamate inhibition revealed that the substitutions of Arg-235 and Glu-264 induced a great loss of the imidazole-carbonyl interaction. However, replacement of Glu 264 with Asp, another acidic amino acid, affected the enzyme function less than the Glu to Gln substitution. In addition, both the Arg-235 and Glu-264 substitutions induced marked increases in the primary isotope effect on the catalysis, suggesting that these amino acids stimulate the hydrogen transfer step in the catalysis. We concluded, therefore, that the guanidino and carboxyl groups of Arg-235 and Glu-264, respectively, cooperatively promote the essential imidazole-substrate interaction, enhancing the substrate binding and catalysis. PMID- 9399586 TI - Functional expression of human mannan-binding proteins (MBPs) in human hepatoma cell lines infected by recombinant vaccinia virus: post-translational modification, molecular assembly, and differentiation of serum and liver MBP. AB - Human mannan-binding proteins (MBPs) occur in two forms, serum MBP (S-MBP) and liver MBP (L-MBP), both of which are synthesized in the liver from a single form of human MBP mRNA. To investigate further the mechanisms of post-translational modification, molecular assembly and differentiation of S-MBP and L-MBP in vitro, we expressed a full-length human MBP cDNA in three human hepatoma cell lines, using the vaccinia virus expression system. The expression of human MBP cDNA reproduced the native MBP differentiation of S-MBP and L-MBP in human hepatoma cells. The recombinant S-MBP was secreted into the medium, and the recombinant L MBP retained in the cells. The former had the ability to activate the complement through the classical or lectin pathway but the latter did not. Furthermore, one notable difference between the two MBPs was the degree of oligomerization through interchain disulfide bonds between subunits. In addition, we showed that both S MBP and L-MBP undergo hydroxylation of lysine and proline residues in collagen like sequences, and that the hydroxylysine is glycosylated to form glucosylgalactosylhydroxylysine (GluGalHyl) and galactosylhydroxylysine (GalHyl). Hydroxylation was required for S-MBP to be assembled into large complexes, the apparent molecular sizes of which were estimated to be 200-1,300 kDa by SDS-PAGE under non-reducing conditions and gel filtration under non-denaturing conditions. The hydroxylation and subsequent glycosylation and oligomerization were inhibited by alpha,alpha'-dipyridyl, an inhibitor of collagen lysyl and prolyl hydroxylases. These results suggested that newly synthesized lectins undergo post translational modifications unique to the two forms of MBP, S-MBP, and L-MBP, in human hepatocytes and hepatoma cells, and that the collagen-like domains of the MBPs play an important role in promoting molecular assembly. PMID- 9399587 TI - Cloning, expression and isoform classification of a minor oleosin in sesame oil bodies. AB - The oil bodies of plant seeds contain a triacylglycerol matrix surrounded by a monolayer of phospholipids embedded with alkaline proteins termed oleosins. Two distinct oleosins are present in the oil bodies of diverse angiosperms, and classified as high and low Mr isoforms according to their relative molecular masses in each species. In sesame oil bodies, besides the two ubiquitous oleosin isoforms (17 and 15 kDa), an additional minor oleosin (15.5 kDa) was revealed on Tricine SDS-PAGE. A full-length cDNA fragment was cloned, sequenced and deduced to be a putative oleosin of 15,446 Da. The gene was constructed in a fusion or non-fusion vector and then over-expressed with different efficiency in Escherichia coli. All three oleosins purified from sesame oil bodies were subjected to immunoassaying using antibodies raised against the over-expressed oleosin. The results confirmed that this gene encodes the sesame 15.5 kDa oleosin. Sequence comparisons with other known oleosins revealed that sesame 15.5 kDa oleosin does not represent a new oleosin isoform class but may have been derived through gene duplication and truncation of sesame 17 kDa oleosin, and possesses the minimal structure of the high Mr oleosin isoform. A conserved amphipathic alpha-helix is predicted in sesame 15.5 kDa oleosin, which may imply a potential biological function associated with this isoform. PMID- 9399588 TI - Structural and mechanistic studies on D-amino acid oxidase x substrate complex: implications of the crystal structure of enzyme x substrate analog complex. AB - As an extension of our recent X-ray crystallographic determination of the tertiary structure of D-amino acid oxidase (DAO) [Mizutani, H. et al. (1996) J. Biochem. 120, 14-17], we solved the crystal structure of the complex of DAO with a substrate analog, o-aminobenzoate (OAB). The alignment between flavin and OAB in the crystal structure of the complex is consistent with charge-transfer interaction through the overlap between the highest occupied molecular orbital of OAB and the lowest unoccupied molecular orbital of flavin. Starting with the atomic coordinates of this complex as the initial model, we carried out molecular mechanics simulation for the DAO-D-leucine complex and thus obtained a model for the enzyme-substrate complex. According to the enzyme-substrate complex model, the alpha-proton is pointed toward N(5) of flavin while the lone-pair of the substrate amino group can approach C(4a) of flavin within an interacting distance. This model as well as DAO-OAB complex enables the evaluation of the substrate-flavin interaction prior to electron transfer from the substrate to flavin and provides two possible mechanisms for the reductive-half reaction of DAO, i.e., the electron-proton-electron transfer mechanism and the ionic mechanism. PMID- 9399589 TI - Functions of characteristic Cys-Gly-His-Cys (CGHC) and Gln-Glu-Asp-Leu (QEDL) motifs of microsomal ER-60 protease. AB - The human ER-60 protease cDNA was expressed in Escherichia coli BL21 (DE3) cells using the pET-20b(+) T7 promoter. The recombinant ER-60 protease was obtained in a water-soluble form and purified through four sequential chromatographies. The ER-60 protease contains two CGHC motifs. When an alanine residue was substituted for the N-terminal cysteine residue in both motifs, the protease activity was not lost. However, when the C-terminal cysteine residue in both motifs was replaced by a serine residue, the cysteine protease activity, which was inhibited by p chloromercuribenzoic acid (pCMB) but not by diisopropyl fluorophosphate (DFP), changed to serine protease activity, which was inhibited by DFP but not by pCMB. These results indicate that the C-terminal cysteine residue(s) of the CGHC motifs may constitute the active site(s) of ER-60 protease. The ER-60 protease has a C terminal QEDL sequence, which was proved to serve as an ER-retention signal by deletion of the QEDL sequence. However, because QEDL could not serve as the ER retention signal for protein disulfide isomerase or ERp72, it is suggested that amino acid residue(s) of ER-60 protease, other than the QEDL sequence itself, is complimentarily responsible for the ER retention of this protein. PMID- 9399590 TI - Methionine aminopeptidase from the hyperthermophilic Archaeon Pyrococcus furiosus: molecular cloning and overexpression in Escherichia coli of the gene, and characteristics of the enzyme. AB - A gene for a methionine aminopeptidase (MAP; EC 3.4.11.18), which catalyzes the removal of amino-terminal methionine from the growing peptide chain on the ribosome, has been cloned from the hyperthermophilic Archaeon, Pyrococcus furiosus, by a novel method effectively using its cosmid protein library, sequenced and expressed in Escherichia coli. The DNA sequence encodes a protein containing 295 amino acid residues with methionine at the N-terminus. From protein analyses of the recombinant protein expressed in E. coli, by using both amino acid sequence analysis from the N-terminus by automated Edman degradation and analyses of molecular masses of the peptides generated by two enzymatic cleavages performed independently, digestions with lysylendopeptidase and Endoproteinase Asp-N, with ionspray mass spectrometry, the primary structure of the protein has been elucidated to be completely identical with that deduced from its DNA sequence. Comparison of the amino acid sequence of P. furiosus MAP (P.f. MAP) with those of other MAPs from Eukarya and Bacteria showed that the protein has a high degree of sequence homology in the stretches surrounding the five cobalt-binding residues fully preserved in all of MAPs determined so far, but P.f. MAP belongs to Type II because it has an extra long insertion of about 60 amino acid residues between the fourth and fifth cobalt-binding ligands, similar to MAPs from human and rat, and to Met-AP2 from Saccharomyces cerevisiae, in comparison to Type I MAPs from Bacteria. Therefore, P.f. MAP seems to be rather close to those from Eukarya, although it is distinct in lacking the N-terminal extension of about 90-150 residues universally found in MAPs from Eukarya. These findings suggest that P.f. MAP is evolutionally located at the Eukarya-Bacteria boundary. The enzyme expressed in E. coli exhibits a considerable thermostability, with a half-life of approximately 4.5 h at 90 degrees C and an optimum temperature of around 90 degrees C. PMID- 9399591 TI - Modification of proteoglycan synthesis by corneal stromal cells on co-culture with either epithelial or endothelial cells. AB - Corneal stromal cells, prepared from 2-day-old chicks and embedded in collagen gel on an insert dish, were co-cultured with corneal epithelial cells or endothelial cells on a fibronectin-coated companion plate. Cell growth and proteoglycan synthesis, as determined as the incorporation of [35S] sulfate and [3H] glucosamine, were examined for each cell type in both combinations. In comparison with single cultures, growth was affected little, while proteoglycan synthesis was appreciably altered in both combinations. Stromal and epithelial cells stimulated proteoglycan synthesis by each other, while endothelial cells stimulated the synthesis by stromal cells, but stromal cells reduced that by endothelial cells to some extent. Endothelial cells alone synthesized proteoglycans much more actively than did epithelial or stromal cells. Analysis of the radiolabeled proteoglycans revealed that endothelial and epithelial cells had different effects on proteoglycan synthesis by stromal cells: the former tended to increase keratan sulfate synthesis by stromal cells, while the latter tended to increase chondroitin sulfate/dermatan sulfate synthesis. Proteoglycan synthesis in the corneal stroma in vivo might thus be controlled by the balance between the antipodal actions of the epithelial and endothelial cell layers. PMID- 9399592 TI - Lipid peroxide overcomes the inability of platelet secretory phospholipase A2 to hydrolyze membrane phospholipids in rabbit platelets. AB - The present study investigated the effect of lipid peroxide on the ability of group IIA secretory phospholipase A2 (IIAsPLA2) to hydrolyze platelet membrane phospholipids. The treatment of rabbit platelets with tert-butyl hydroperoxide (BHP) and FeSO4 generated malondialdehyde, an index of lipid peroxidation, and slightly induced arachidonic acid liberation and lysophosphatidylcholine formation. Further addition of IIAsPLA2 purified from rabbit platelets synergistically enhanced the liberation and the formation induced by the oxidizing reagents, although the enzyme alone did not. When the IIAsPLA2 was pretreated with heparin, the enhancement was not observed. The combination of IIAsPLA2 with linoleic acid hydroperoxide and FeSO4 also caused synergistic arachidonic acid liberation. Furthermore, IIAsPLA2 enhanced thromboxane B2 generation and platelet aggregation induced by BHP and FeSO4. The synergistic aggregation was sensitive to indomethacin. With a membrane fraction as a substrate, IIAsPLA2 caused arachidonic acid liberation, which was enhanced in the presence of BHP and FeSO4. These results suggest that modification of membrane phospholipids by oxidizing reagents increases the accessibility of the membrane to platelet IIAsPLA2, and sequential enhancement of arachidonic acid liberation may contribute to the propagation of oxidative stress-induced cellular injury. PMID- 9399593 TI - Identification of endogenous substrates for Drosophila calpain from a salt extracted fraction of Drosophila ovaries. AB - Drosophila calpain (Dm-calpain) produced in Escherichia coli has a distinct Ca2+ dependent activity. By using a recombinant Dm-calpain, we searched for its substrates occurring in Drosophila ovaries, where Dm-calpain is expressed. Among a number of major proteins, several proteins in a salt-extracted fraction were selectively degraded by Dm-calpain in a Ca2+-dependent manner. The major substrates were identified by microsequencing the lysylendopeptidase-digested proteins. Three ribosomal proteins, the L5, L7, and L8 subunits of the 60S ribosome, were found to be potential Dm-calpain substrates. In addition, the alpha subunit of elongation factor-1 (EF-1alpha), a multi-functional protein involved in both protein synthesis and cytoskeletal regulation, was shown to be cleaved by Dm-calpain into several distinct fragments when expressed as a GST fusion protein. Endogenous EF-1alpha in ovary extracts was also shown by western blot analysis to be similarly degraded. These observations suggest that Dm calpain may regulate protein synthesis and cytoskeletal structure through its degradative or processing activity. PMID- 9399594 TI - Isolation of a novel human gene from the Down syndrome critical region of chromosome 21q22.2. AB - Down syndrome is the most common birth defect, and is caused by trisomy 21. We identified a novel gene in the so-called Down syndrome critical region by means of computer-aided exon prediction and subsequent cDNA cloning. The gene, designated as DCRA (Down syndrome Critical Region gene A), consists of eight exons of 3,252 bp in total and encodes a large open reading frame of 297 amino acid residues. The open reading frame shows significant homology to Hbeta58, a mouse gene essential for embryogenesis, PEP8, a yeast homologue of Hbeta58, and an expressed sequence tag of Arabidopsis thariana, suggesting that DCRA has some important function that has been conserved during the course of evolution. DCRA is expressed in most tissues examined, including fetal and adult brain, heart, lung, liver, and kidney. The cDNA of the DCRA mouse homologue, Dcra, was also cloned. It is 2,157 bp long and has an open reading frame of 297 amino acid residues, which shows 92% identity to human DCRA. Dcra is expressed in all the embryo and adult tissues examined. PMID- 9399595 TI - Identification of the nucleotides in the A-rich bulge of the Tetrahymena ribozyme responsible for an efficient self-splicing reaction. AB - P5abc is a large extension of the P5 element characteristic of subclasses IC1 and IC2 of group I introns. It has a conserved region termed the A-rich bulge, that is responsible for activation of the Tetrahymena self-splicing intron. By employing a modified color-colony assay system, we identified four adenosines in the bulge that are responsible for an efficient splicing reaction. On comparison with the X-ray crystal structure of the P4-5-6 domains of the Tetrahymena intron, three adenosines at positions 183, 184, and 186 were found to be identical to those significantly contributing to the formation of its tertiary structure. However, our results show that an adenosine at 187 is involved in the formation of a Watson-Crick base pair with U135, although it forms a Hoogsteen base pair in the crystal structure. PMID- 9399596 TI - Analysis of the substrate binding site and carboxyl terminal region of vacuolar H+-pyrophosphatase of mung bean with peptide antibodies. AB - Vacuolar H+-translocating inorganic pyrophosphatase is a single-protein enzyme and uses a simple substance as an energy donor. Functional domains of the enzyme were investigated by using antibodies specific to peptides corresponding to the putative substrate-binding site (DVGADLVGKVE) in the hydrophilic loop and the carboxyl terminal part. The antibody to the former peptide clearly reacted with the pyrophosphatases of different plant species, and strongly inhibited the hydrolytic activity of the purified enzymes and the proton pumping activity of membrane vesicles. These results indicate that the sequence functions as an actual substrate-binding site and is a common motif. The antibody to the carboxyl terminal part reacted only to the mung bean enzyme, suppressing its hydrolytic and proton pumping activities. The results suggest that the carboxyl terminus is exposed to the cytosol and is close to the catalytic site. H+-Pyrophosphatase hydrolyzed triphosphate and tetraphosphate at low rates. Phytic acid, myo inositol hexaphosphate, inhibited the enzyme even in the presence of Mg2+. The concentration for 50% inhibition was 0.15 mM. The inhibition of H+-PPase by dicyclohexyldiimide was partly reversed by Mg2+. The catalytic site and the membrane topology of the enzyme are discussed. PMID- 9399597 TI - Is the cannabinoid CB1 receptor a 2-arachidonoylglycerol receptor? Structural requirements for triggering a Ca2+ transient in NG108-15 cells. AB - The effects of delta9-tetrahydrocannabinol and 2-arachidonoylglycerol on the intracellular free Ca2+ concentration ([Ca2+]i) in NG108-15 cells were examined in detail. We found that delta9-tetrahydrocannabinol induces a rapid, modest increase in [Ca2+]i. The response was detectable with 3 nM delta9 tetrahydrocannabinol. We also found that very low concentrations of 2 arachidonoylglycerol elicit a rapid, more prominent increase in [Ca2+]i. Such a response was observed not only in NG108-15 cells but also in N18TG2 cells. The response induced by 2-arachidonoylglycerol in either NG108-15 cells or N18TG2 cells was abolished by pretreatment of the cells with a cannabinoid CB1 receptor specific antagonist, SR141716A, suggesting that 2-arachidonoylglycerol interacts with the CB1 receptor to induce the response. The results of an experiment involving a phospholipase C inhibitor suggested that phospholipase C is involved in the rapid increase in [Ca2+]i induced by 2-arachidonoylglycerol. We also found that 1(3)-arachidonoylglycerol exhibits similar activity to that of 2 arachidonoylglycerol, although its activity at low concentrations was somewhat weak compared with that of 2-arachidonoylglycerol. We further confirmed that several structural analogues of 2-arachidonoylglycerol were less active compared with 2-arachidonoylglycerol. These results suggest that the structure of 2 arachidonoylglycerol is strictly recognized by the CB1 receptor, which raises the possibility that the CB1 receptor is originally a 2-arachidonoylglycerol receptor. PMID- 9399598 TI - Sparfloxacin: potential clinical and economic impact in the treatment of respiratory infections. AB - Sparfloxacin is a new oral fluoroquinolone antimicrobial that is highly active against common respiratory pathogens, including multiresistant strains. It is well absorbed and has excellent penetration into upper and lower respiratory tissues. Sparfloxacin is administered once a day and does not interfere with the metabolism of other drugs. The agent is highly effective and safe in the treatment of acute sinusitis, exacerbations of chronic bronchitis, and community acquired pneumonia. Due to its activity against multidrug-resistant respiratory pathogens, it has the potential to prevent hospitalization as well as decrease parenteral antibiotic therapy. Consequently, it may generate significant pharmacoeconomic benefits to patients and payers of medical care. PMID- 9399599 TI - Sirolimus, a new, potent immunosuppressive agent. AB - Sirolimus (SRL), a potent immunosuppressant, is currently being investigated in phase III trials for prophylaxis of renal transplant rejection. The mechanism of action of SRL is a blockade of the response of T and B cells to cytokines, thereby preventing cell cycle progression in G1 and consequently cell proliferation. There seems to be a good correlation between SRL concentrations, estimated as exposure by the area under the concentration versus time curve, and trough whole blood concentration, so that therapeutic monitoring may be done by trough levels only. Because of the low frequency of allograft rejection, there has been no documented correlation between trough concentrations and efficacy. Trough SRL concentrations of 15 ng/ml or higher seem to be associated with an increased frequency of adverse effects. Drug-associated toxicities include thrombocytopenia, leukopenia, and increases in cholesterol and triglyceride levels. The drug has synergy with cyclosporine (CsA) in vitro as well as in animal and clinical studies. In phase II trials the combination of SRL-CsA therapy reduced the frequency of acute rejection episodes, permit withdrawal of concomitant corticosteroid therapy, and allowed reduction of CsA dosages in nonblack patients. PMID- 9399600 TI - Atorvastatin calcium: an addition to HMG-CoA reductase inhibitors. AB - Atorvastatin calcium is an HMG-coenzyme A (CoA) reductase inhibitor that was approved by the Food and Drug Administration on December 17, 1996. Like other such agents, it inhibits the action of HMG-CoA reductase and thereby decreases endogenous cholesterol synthesis, leading to a decrease in circulating low density lipoprotein cholesterol. In addition to its effect on lipoprotein profile, atorvastatin reduces triglycerides to a greater extent than other HMG CoA reductase inhibitors. These actions occur in a dose-dependent fashion. The adverse effect profile is similar to that of other agents in this class. Indications for atorvastatin include primary hypercholesterolemia as well as other lipid disorders. PMID- 9399601 TI - Mycophenolate mofetil. AB - Mycophenolate mofetil is the morpholinoethylester prodrug of mycophenolic acid, an agent that inhibits the proliferation of B and T lymphocytes through noncompetitive, reversible inhibition of inosine monophosphate dehydrogenase, a key enzyme in the de novo synthetic pathway of guanine nucleotides. Currently, mycophenolate mofetil is approved for the prevention of acute renal allograft rejection when given in combination with cyclosporine and steroids. Several studies also demonstrated that the agent is effective in the treatment of refractory rejection in renal, heart, and liver transplant recipients, and may have efficacy in the treatment of chronic rejection as well. PMID- 9399602 TI - New drug interactions with zidovudine. AB - Polypharmacy is commonly encountered in human immunodeficiency virus (HIV) positive patients, and the risk and frequency of drug-drug interactions are significant in this patient population. Most HIV-positive patients receive the antiretroviral drug zidovudine (3'-azido-3'-deoxythymidine, ZDV), the first drug to be approved for the treatment of HIV. Many drug interactions with ZDV have already been reported. As HIV pharmacotherapy becomes more complex, the potential for drug-drug interactions is likely to increase significantly. PMID- 9399603 TI - A systematic review and meta-analysis of the incidence of cancer in randomized, controlled trials of verapamil. AB - We conducted a systematic review of all published randomized, controlled trials to assess the risk of cancer or death in patients receiving verapamil for hypertension, angina pectoris, or cardiac arrhythmias. Meta-analysis comparing the risk of new cancers, cancer deaths, and all deaths was performed. Thirty-nine trials comprising 11,201 patients were eligible. Study durations ranged from 8 days-6 years (mean 29.5 wks). Nine trials (6507 patients) were 24 weeks in duration or longer. For cancer and cancer death, OR was 1.20 (95% CI = 0.60-2.42) for verapamil versus active controls and 0.73 (95% CI = 0.39-1.39) for verapamil versus placebo. For all deaths, OR was 1.13 (95% CI = 0.70-1.82) for verapamil versus active controls and 0.85 (95% CI = 0.71-1.00) for verapamil versus placebo. Sensitivity analysis for the 9 trials 24 weeks' duration or longer gave similar results. There is no statistically significant increased risk of cancer or deaths with verapamil compared with active controls or placebo. PMID- 9399604 TI - Drug-induced neuromuscular blockade and myasthenia gravis. AB - Myasthenia gravis is an uncommon disorder of the neuromuscular junction resulting in weakness of all striated voluntary muscles. Therapeutic advances have increased patients' age and survival. Older patients with myasthenia gravis may have additional medication needs. Numerous drugs have experimental and clinical evidence of neuromuscular blockade. A MEDLINE search of the English literature from 1966 to the present pertinent to drug-induced myasthenia gravis was performed. Additional literature was obtained from reference citations of relevant articles. Drugs with several reports of neuromuscular blockade were assessed for causality by a recognized probability scale. Prednisone was most commonly implicated as aggravating myasthenia gravis, and D-penicillamine was most commonly associated with myasthenic syndrome. The greatest frequency of drug induced neuromuscular blockade was seen with aminoglycoside-induced postoperative respiratory depression. However, drugs most likely to impact myasthenic patients negatively are those used in the treatment of the disease. These include overuse of anticholinesterase drugs, high-dose prednisone, and anesthesia and neuromuscular blockers for thymectomy. PMID- 9399605 TI - Is oral sotalol effective in converting atrial fibrillation to sinus rhythm? AB - d,l-Sotalol is a noncardioselective beta-blocker that has class III antiarrhythmic activity. It is often used to convert atrial fibrillation (AF) to normal sinus rhythm. Since class III agents increase action potential duration and refractoriness in atrial tissue without affecting conduction, they are theoretically considered ideal agents for the treatment of reentrant arrhythmias such as AF. We reviewed the literature evaluating the efficacy of sotalol for restoring sinus rhythm in patients with acute or chronic AF. Articles indexed on MEDLINE (1966-1996) and referenced articles not identified by MEDLINE that compared sotalol with placebo or another antiarrhythmic agent were included. Sotalol was significantly inferior to quinidine in converting AF of recent onset (< 48 hrs) to sinus rhythm. In patients with duration of AF of more than 48 hours, sotalol was significantly less effective than quinidine and comparable with placebo. Conversion rates for sotalol in all studies combined ranged from 8 49%. Published studies do not support the drug for conversion of AF to sinus rhythm. Larger well-designed studies are required to evaluate its efficacy and optimum dosage for this indication. Until further data are available, pharmacologic cardioversion with traditional class I antiarrhythmic agents may be preferable as they are effective particularly for recent-onset AF. PMID- 9399606 TI - Comparison of intravenous diltiazem and verapamil for the acute treatment of atrial fibrillation and atrial flutter. AB - STUDY OBJECTIVES: To compare the efficacy and safety of intravenous diltiazem and verapamil in controlling ventricular rate in patients with atrial fibrillation or flutter, and to evaluate the effects of these agents on left ventricular systolic function. DESIGN: Prospective, randomized, double-blind, crossover study. SETTING: University-affiliated hospital and Veterans Administration hospital. PATIENTS: Seventeen men with atrial fibrillation or flutter with a ventricular rate of 120 beats/minute or higher and a systolic blood pressure of 100 mm Hg or greater. INTERVENTIONS: Patients received up to two intravenous boluses of either diltiazem or verapamil, followed by an 8-hour continuous infusion if a therapeutic response was achieved (phase I). After a washout period, patients who responded were crossed over to receive the other drug in a similar fashion (phase II). MEASUREMENTS AND MAIN RESULTS: At the end of each infusion, the patient's ejection fraction was assessed by gated angiography. Of the 17 men initially randomized, 8 successfully completed both phases I and II. In these patients, baseline mean (+/- SD) ventricular rates before treatment with intravenous diltiazem and verapamil were 138 +/- 15 and 132 +/- 9 beats/minute, respectively (NS). At 2 minutes after the initial bolus dose, the mean ventricular rate decreased to 100 +/- 13 beats/minute in the diltiazem group compared with 114 +/- 17 beats/minute in those receiving verapamil (p < 0.05). Mean ventricular rates of 96 +/- 11 and 97 +/- 9 beats/minute were maintained during the 8-hour continuous infusion of diltiazem and verapamil, respectively (NS). On completion of the bolus dose(s) and during continuous infusions, there were no significant differences in blood pressures between the groups. Mean ejection fractions were 35.6 +/- 13.6% and 35.5 +/- 15.4% in the diltiazem and verapamil groups, respectively (NS). For the 17 patients, the mean maximum percentage decreases in blood pressure were not significantly different between groups. However, three patients developed symptomatic hypotension, all of whom were randomized to receive verapamil initially. CONCLUSION: Intravenous diltiazem and verapamil are comparable in terms of efficacy and effect on systolic function in patients with rapid atrial fibrillation and flutter. However, hypotension may limit therapy with verapamil in some patients. PMID- 9399607 TI - Cisplatin nephrotoxicity: a multivariate analysis of potential predisposing factors. AB - STUDY OBJECTIVE: To evaluate the usefulness of biologic and pharmacologic parameters for early identification of cisplatin-induced renal dysfunction. DESIGN: Prospective evaluation of 62 consecutively admitted patients with cancer. SETTING: Cancer center. PATIENTS: Sixty-two consecutive patients with cancer (52 men, 10 women; mean age 61.9 yrs). INTERVENTIONS: Patients received cisplatin as a single short intravenous infusion every 3 weeks. One hundred twenty-one cycles were analyzed. The dosage in the first cycle ranged between 61 and 105 mg/m2 (mean 84 mg/m2). All patients received a standard hydration protocol. MEASUREMENTS AND MAIN RESULTS: Renal function was evaluated for each cycle before treatment (day 0) and before next cycle (day 21) based on the estimated creatinine clearance (Clcr). For each cycle, the weighted relative decrease (WD) of Clcr was calculated (WDClcr = 100 x [Clcr (day 0) - Clcr (day 21)]/[Clcr (day 0)](2). Total and ultrafilterable (UF) platinum were measured as a single-sample assay taken 16 hours after the end of cisplatin administration. The mean WDClcr was 0.07 min/100 ml (range -1.0 to +1.7 min/100 ml). The intensity of renal dysfunction evaluated by WDClcr was independent of cisplatin dosage, age, sex, body surface area, initial Clcr, and cycle number. Of interest, total and UF platinum concentrations were significantly correlated to WDClcr: the higher the platinum concentration, the greater the intensity of renal dysfunction. In stepwise regression analysis, UF platinum concentration was the only selected factor. The best prediction of UF platinum was obtained by stepwise regression including cisplatin dosage, initial Clcr, and cycle number (r=0.58, p<0.0001). CONCLUSION: We consider our results to be a first step toward a clinical strategy to identify patients at risk for renal dysfunction after cisplatin treatment. PMID- 9399609 TI - Evidence for negative feedback of extracellular methotrexate on blasts of acute lymphoblastic leukemia in vitro. AB - STUDY OBJECTIVE: To explore the value of high-dose methotrexate (MTX). SUBJECTS: Blast cells from 15 patients with acute lymphoblastic leukemia. INTERVENTIONS: We compared uptake and polyglutamation of [3H]-MTX by freshly isolated leukemic blasts in vitro after 24-hour exposure to 1, 10, and 50 microM [3H]-MTX. MEASUREMENTS AND MAIN RESULTS: Mean MTX uptake (pmol/10(6) cells) was 0.78 +/- 0.19, 2.3 +/- 0.54, and 5.9 +/- 1.9, respectively, and mean polyglutamation was 82%, 66%, and 46%. Consequently, mean MTX polyglutamates were 0.68 +/- 0.18, 1.5 +/- 0.47, and 2.2 +/- 0.67 pmol/10(6) cells. Three of 15 patient samples had no detectable polyglutamation of MTX at 50 microM but MTX polyglutamates were detectable at 1 microM. Two of these three had a decrease in MTX polyglutamates at 10 versus 1 microM. In eight precursor B cell samples there was a significant difference in median MTX polyglutamates at 1 versus 10 microM but not 10 versus 50 microM. CONCLUSION: Increasing extracellular MTX concentrations may be counterproductive for some patients with acute lymphoblastic leukemia. If MTX polyglutamates are important for efficacy, optimal delivery of MTX may have to be determined by individual metabolism rather than by targeting a specific drug concentration. PMID- 9399608 TI - Long-acting diltiazem CD is safe and effective in a hypertensive Mexican-American population. AB - STUDY OBJECTIVE: To evaluate the safety and effectiveness of diltiazem CD for reducing blood pressure in Mexican-American patients with mild to moderate hypertension. DESIGN: Randomized, double-blind, placebo-controlled trial. SETTING: Twelve clinical sites in the United States. PATIENTS: Patients with baseline diastolic blood pressures between 95 and 115 mm Hg. INTERVENTIONS: Patients were treated with an average daily dose of diltiazem CD 246 mg (60 patients) or placebo (58 patients) to achieve a trough diastolic blood pressure below 90 mm Hg. MEASUREMENTS AND MAIN RESULTS: Diltiazem CD significantly reduced mean diastolic blood pressure compared with placebo, -8.2 versus -4.1 mm Hg, respectively (p=0.0025). Diastolic blood pressure below 90 mm Hg or a reduction of 10 mm Hg or more was achieved by 57% of diltiazem CD versus 28% of placebo recipients. Systolic blood pressure and heart rate were also reduced with diltiazem CD. Adverse events were mild, with similar frequency for diltiazem CD (15%) and placebo (19%). CONCLUSION: Diltiazem CD is safe and effective in hypertensive Mexican-Americans, and diastolic blood pressure reductions compare with those in non-Hispanic white patients. PMID- 9399610 TI - Lidocaine does not affect myocardial electrical heterogeneity: implications for low proarrhythmic actions. AB - An area of unidirectional conduction block is one requirement for reentrant arrhythmias to occur. Functional block caused by dispersion of repolarization and refractoriness is the most probable mechanism of drug-induced unidirectional conduction block. We assessed the effects of lidocaine on spatial dispersion of myocardial repolarization and refractoriness in the intact porcine heart. Monophasic action potential duration at 90% repolarization, effective refractory period (ERP), and ventricular fibrillation cycle length (VFCL) were measured at two endocardial and one epicardial sites at baseline and during a treatment phase with D5W (n=11) or lidocaine 10 mg/kg/hour (n=12). Dispersion was calculated as the difference between the maximum and minimum values of the three recording sites. Lidocaine produced significant changes in ERP, VFCL, paced QRS duration, and intraventricular conduction time. It did not change basal levels of dispersion in repolarization and refractoriness. Lidocaine produced changes in myocardial electrophysiology that are uniform across the myocardium and thus did not change myocardial electrical heterogeneity. This may be a mechanism of the agent's lower proarrhythmic effects compared with other sodium channel blockers that increase myocardial electrical heterogeneity. PMID- 9399611 TI - Evaluation of hypertensive patients after care provided by community pharmacists in a rural setting. AB - We evaluated blood pressure control, quality of life, quality of care, and satisfaction of patients who were monitored by specially trained community pharmacists in a group medical practice. After participating in an intensive skill development program, pharmacists performed in an interdisciplinary team in a rural clinic. The primary objective was assessed by evaluating outcome variables at 6 months compared with baseline in 25 patients randomly assigned to a study group. A control group of 26 patients was also evaluated to determine if outcome variables remained constant from baseline to 6 months. Systolic blood pressure was reduced in the study group (151 mm Hg baseline, 140 mm Hg at 6 mo, p<0.001) and diastolic blood pressure was significantly lower at 2, 4, and 5 months compared with baseline. Ratings from a blinded peer review panel indicated significant improvement in the appropriateness of the blood pressure regimen, going from 8.7 +/- 4.7 to 10.9 +/- 4.5 in the study group (p<0.01), but they did not change in the control group. Several quality of life scores improved significantly in the study group after 6 months (p<0.05). These included physical functioning (61.6 vs 70.7), physical role limitations (56.8 vs 72.8), and bodily pain (60.0 vs 71.7) at baseline and 6 months, respectively. There were no significant changes in the control group. Patient satisfaction scores were consistently higher in the study group at the end of the study. Our results indicate that when community pharmacists in a clinic setting are trained and included as members of the primary care team, significant improvements in blood pressure control, quality of life, and patient satisfaction can be achieved. PMID- 9399612 TI - Cost analysis of the American College of Chest Physicians guidelines for deep vein thrombosis prophylaxis in patients undergoing orthopedic arthroplastic surgery. AB - Guidelines for prophylaxis of deep vein thrombosis secondary to orthopedic surgery have been developed. In selecting a specific drug for formulary inclusion, it is ideal for an individual institution to determine the cost of therapy, as well as the frequency of adverse events and the cost of treating them for each agent undergoing consideration. Cost-effectiveness analysis using incremental cost-effectiveness ratios and sensitivity analyses are useful for determining which drug may be most cost effective. PMID- 9399613 TI - Effect of infusing fat emulsion into extracorporeal membrane oxygenation circuits. AB - Our objectives were to identify problems associated with the administration of fat emulsion by extracorporeal membrane oxygenation (ECMO) circuits, and gather information from other institutions on standards of practice and the complications associated with infusion of fat emulsion by ECMO to infants and children. In vitro analysis was performed using six circuits. Fat emulsion was infused into a prereservoir port at 3 ml/hour. Circuits and blood samples collected distal to the oxygenator were inspected visually for layering (separation of fat emulsion from blood), agglutination, and phase separation (formation of an oil layer) at 0.5, 1, 2, 4, 6, 12, and 24 hours. At 24 hours, samples were reevaluated and circuits dissected. All circuits showed layering and agglutination. Blood clots were present in five circuits during the simulation. There was no evidence of phase separation in the samples. Adhesion of emulsion to the equipment was present in all circuits. Five of the membrane oxygenators contained clots. One contained long strands of fat; separation of its mesh revealed an oily residue indicating disruption of the stability of the emulsion. Survey responses from 54 centers found that 78% used fat emulsion routinely in neonatal or pediatric patients receiving ECMO. Most used both ECMO and separate venous access for the infusion, depending on availability. Twenty-two (52%) of the 42 centers using fat emulsion had a policy in place regarding site selection. Of those, 73% preferred central venous access, another 18% used a prereservoir port of the ECMO circuit. The most frequently reported problems with administration through the circuit were cracking of stopcocks, clogging and malfunction of the membrane oxygenator, agglutination of the emulsion, and increase in blood clot formation. Our results suggest that fat emulsion should be infused through a separate intravenous site whenever possible. Based on these results and current practices of most ECMO centers, a clinical trial is currently being conducted to provide additional information. PMID- 9399614 TI - Experience with crystalline niacin as the preferred drug for dyslipidemia in a specialty clinic. AB - To determine the tolerability and efficacy of crystalline niacin in reaching target lipid goals, we conducted a retrospective review of medical records of 62 patients treated with the agent over 2 years in a lipid clinic at a nonacademic veterans hospital. Most patients received niacin for hypercholesterolemia. Thirty one patients (50%) stopped therapy due to adverse events, principally, intolerable cutaneous reactions. Twenty-nine withdrew from therapy during the first 6 weeks of treatment. Of those who tolerated niacin, 23 did not achieve target lipid serum concentrations at the maximum tolerated dosage; 8 did achieve target concentrations. Thus crystalline niacin was largely ineffective in treating patients with dyslipidemia. PMID- 9399615 TI - N-desmethylclozapine, an insensitive marker of clozapine-induced agranulocytosis and granulocytopenia. AB - We reviewed the charts of 58 patients with treatment-refractory schizophrenia who were receiving clozapine, to determine if the drug's active metabolite, N desmethylclozapine, is a biologic marker for impending clozapine-induced granulocytopenia and agranulocytosis. No significant correlation between granulocyte counts and patient demographic variables of clozapine and N desmethylclozapine steady-state plasma concentrations, clozapine:N desmethylclozapine ratio, age, gender, clozapine dosage, smoking status, and race were found. We believe N-desmethylclozapine is not a clinically useful marker for monitoring the effect of clozapine on granulocyte integrity. On the contrary, its plasma concentrations correlated positively with granulocyte counts. PMID- 9399616 TI - Impact of CYP2D6 poor metabolizer phenotype on propranolol pharmacokinetics and response. AB - We conducted an open-label study to determine the impact of cytochrome P-4502D6 (CYP2D6) on propranolol pharmacokinetics and response in 12 healthy men with CYP2D6 extensive metabolizer (EM) phenotype and 3 healthy men with CYP2D6 poor metabolizer (PM) phenotype. Subjects received R,S-propranolol hydrochloride 80 mg every 8 hours for 16 doses. After the sixteenth dose, blood and urine samples were collected for 24 hours, and serum propranolol and urine metabolite concentrations were determined by chiral high-performance liquid chromatography. Heart rate response to treadmill exercise was measured serially over 24 hours. Apparent oral clearance of propranolol and partial metabolic clearance values of propranolol to 4-hydroxypropranolol (HOP), propranolol glucuronide, and naphloxylactic acid (NLA) were estimated. Apparent oral clearance and elimination half-life of propranolol were not different between EMs and PMs. Partial metabolic clearance of propranolol to HOP was significantly higher and to NLA was significantly lower in EMs than in PMs. No differences in percentage reductions in exercise heart rate were observed between EMs and PMs. The CYP2D6 PM phenotype has no effect on propranolol blood concentrations and does not alter response to propranolol. Our data also suggest that CYP2D6 mediates approximately 65% and 70% of S- and R-propranolol's 4-hydroxylation, respectively. PMID- 9399617 TI - Paradoxic excitation with diphenhydramine in an adult. AB - Although paradoxic excitation may rarely occur in children and adults with conventional dosages of antihistamines, few case reports have appeared in the literature. We cared for a 46-year-old patient who became extremely agitated after receiving a dose of intravenous diphenhydramine. PMID- 9399618 TI - Elevation of the erythrocyte sedimentation rate precedes exacerbation of bleomycin-induced pulmonary toxicity: report of two cases and review of literature. AB - Bleomycin is included in a number of potentially curative chemotherapy regimens. It is associated with distinct forms of pulmonary toxicity, with interstitial pneumonitis the most common. Early detection of pulmonary toxicity is not always predictable by monitoring serial chest radiographs and pulmonary function tests. Even newer serum markers are not useful indicators of bleomycin-induced pulmonary damage. Two patients developed bleomycin pulmonary toxicity, in both of whom increases in erythrocyte sedimentation rate (ESR) preceded clinical deterioration and radiographic changes. The ESR may have potential significance as a monitoring test in patients receiving bleomycin. PMID- 9399619 TI - Cross-resistance to both atracurium- and vecuronium-induced neuromuscular blockade in a critically ill patient. AB - A previously healthy 33-year-old woman received neuromuscular blocking agents during treatment of severe adult respiratory distress syndrome secondary to pneumococcal pneumonia and septic shock. Atracurium infusion rates were progressively increased, preceded by repeated loading doses up to a maximum of 3.57 mg/kg/hour, but produced inadequate neuromuscular blockade as assessed by clinical and ventilatory parameters as well as train-of-four (TOF) monitoring. Atracurium was discontinued and vecuronium infusions of 2.3 mg/kg/hour finally produced adequate paralysis for 7 days. Increasing vecuronium requirements then prompted discontinuation of neuromuscular blockade. Atracurium was reinstituted 2 days later because of worsening pulmonary function. Infusion rates of 3.04 mg/kg/hour were again required, together with high-dose midazolam and fentanyl, to achieve adequate oxygenation with acceptable airway pressures; however, TOF monitoring showed an unacceptable level of paralysis. Cross-resistance among chemically dissimilar neuromuscular blocking agents poses a difficult patient management problem and supports a pharmacodynamic basis of resistance to these agents. PMID- 9399620 TI - Bradycardia associated with intravenous administration of tacrolimus in a liver transplant recipient. AB - Cardiotoxicity due to tacrolimus is documented infrequently in the medical literature. Sinus bradycardia associated with intravenous tacrolimus occurred in a 15-year-old orthotopic liver transplant recipient. The mechanism of this adverse effect is unknown; however, it does not appear to be concentration dependent, and in this patient it resolved on changing to oral therapy. Practitioners should be aware that intravenous administration of tacrolimus may be associated with adverse cardiac events including sinus bradycardia. PMID- 9399621 TI - Warfarin resistance associated with intravenous lipid administration: discussion of propofol and review of the literature. AB - Intravenous lipids are often required for parenteral nutritional (PN) support in critically ill patients and are administered with continuous sedation if patients are receiving propofol, which contains soybean oil 10% as an emulsified preparation. High-dose propofol infusion was associated with reversal of enteral and intravenous warfarin anticoagulation in a 39-year-old woman with severe Crohn's disease. Despite increasing the daily dose of warfarin to 30 mg, anticoagulation was not achieved until propofol was discontinued. Reversal of anticoagulation recurred when PN support was supplemented with Liposyn II 20%. Lipid emulsions may interfere pharmacodynamically with warfarin activity by enhancing the production of clotting factors, facilitating platelet aggregation, or supplying vitamin K. They also may facilitate warfarin binding to albumin. Until further information regarding the mechanism of interference is elucidated, heparin therapy should be considered for initial anticoagulation in patients with intestinal absorptive deficiencies who receive high-dose lipid emulsions and require reliable anticoagulation. If warfarin is given, the international normalized ratio should be monitored daily to ensure adequate anticoagulation. PMID- 9399622 TI - Prolonged hypoglycemic crisis associated with glyburide. AB - An elderly patient taking glyburide 5 mg/day for noninsulin-dependent diabetes mellitus was hospitalized because of severe hypoglycemia. Laboratory results indicated both renal and hepatic abnormalities were present at the time of admission. Despite infusion of 10% dextrose and supplemental boluses of 50% dextrose, the hypoglycemic crisis persisted for 3 days. After it resolved, the patient's diabetes was controlled by diet alone. The patient's age and the presence of hepatic and/or renal impairment must be taken in account in prescribing glyburide. Recognizing patients who may require dosage changes, and educating them on the signs and symptoms of hypoglycemia, may help prevent hospitalizations resulting from this complication associated with glyburide. PMID- 9399623 TI - Delayed hypersensitivity reaction to vancomycin. AB - The use of vancomycin has increased dramatically in recent years with the emergence of methicillin-resistant Staphylococcus aureus and coagulase-negative staphylococci as important hospital pathogens. A patient receiving vancomycin 1.75 g intravenously as a single daily dose developed a syndrome characterized by high fever, erythema multiforme, eosinophilia, and presumed interstitial nephritis. This delayed hypersensitivity reaction resolved with discontinuation of the drug and treatment with methylprednisolone. PMID- 9399625 TI - A simpler approach to pharmacokinetic dosage adjustments. PMID- 9399624 TI - Diltiazem-associated thrombocytopenia. AB - Nine days after starting diltiazem therapy for new-onset atrial fibrillation, a patient's platelet count had diminished to 61 x 10(3)/mm3, and 2 weeks later the nadir was reached at 23 x 10(3)/mm3. No hypersensitivity reaction otherwise was noted, and other hematologic values were unaffected. The patient's platelet counts were not affected by platelet transfusions. Bone marrow examination showed normal to increased numbers of megakaryocytes, suggesting peripheral destruction. After discontinuing diltiazem and administering high-dose immunoglobulin infusions, the platelet count returned to normal. This case suggests an immune mediated cause for thrombocytopenia after exposure to diltiazem. PMID- 9399626 TI - The role of academia in community-based pharmaceutical care. AB - Developing models of pharmaceutical care (PC) for educating students and practitioners represents a fundamental role for schools of pharmacy. Virginia Commonwealth University has sought to facilitate the implementation of PC in the community by hiring faculty to practice in this setting. The mission of the faculty is to implement PC in a community pharmacy practice, to develop clerkship sites for Pharm.D. students, and to evaluate the impact of PC services in the community. Examples of an independent pharmacy model, a grocery chain model, and a retail chain model of care may serve a dual purpose for faculty members, that is, define responsibilities for the academic institution and for the community practice environment. PMID- 9399627 TI - CCK biosynthesis and processing: recent progress and future challenges. AB - The peptide cholecystokinin (CCK), like other peptides which pass through the regulated secretory pathway, undergoes a number of post-translational modifications during its biosynthesis including tyrosine sulfation, endoproteolytic cleavage, and trimming by carboxypeptidases. This minireview summarizes what is known about this process in endocrine cells and in the Cpe(fat)/Cpe(fat) mouse and points out what challenges remain for future research. PMID- 9399628 TI - Effect of high salt intake on plasma and tissue concentration of endogenous ouabain-like substance in the rat. AB - The effect of high salt intake on serum concentration and tissue distribution of ouabain-like substance (OLS) was examined in rats. Sprague-Dawley rats (n=8) were placed on a high salt diet by the inclusion of 1.8% sodium chloride in drinking water for 7 days and a 'control' group (n=8) was maintained on normal drinking water during the study period. Serum and tissue OLS was measured by radioimmunoassay after solid phase extraction. High salt intake significantly increased serum OLS concentration (1.43 +/- 0.06 vs 1.14 +/- 0.05 nmol/L; mean +/ SEM, P=0.002). In both groups, the adrenal showed significantly (p < 0.001) higher OLS content compared to liver, kidney, heart and brain. HPLC of rat serum extract resolved a major peak with a retention time identical to that of standard ouabain, further confirming the nature of OLS. We conclude that high salt intake increases endogenous production of OLS, which appears to originate from the adrenal gland in the rat. PMID- 9399629 TI - NADPH-diaphorase containing cells and fibers in the central nervous system of squid, Loligo bleekeri keferstein. AB - Distribution of NADPH-diaphorase in the central nervous system of squid was determined using histochemical technique. We found NADPH-diaphorase positive cell bodies and fibers both in the optic and the posterior anterior lobe and fibers in the peduncle lobe. These results clarify the biochemical similarity between two structurally similar organs of invertebrate and vertebrate, the peduncle lobe and the anterior basal lobe, and the cerebellum. NADPH-diaphorase positive fibers innervated the inner granule layer and the outer plexiform layer of the outer cortex of the optic lobe. This is in good agreement with avian centrifugal projection from isthmo-optic nucleus to retina where nitric oxide synthase is known to be contained. There may be at least two distinct neural systems, the motor control system and the visual information processing system, which use nitric oxide as a transmitter or modulator in the squid central nervous system. PMID- 9399630 TI - Metabolism of haloperidol to pyridinium species in patients receiving high doses intravenously: is HPTP an intermediate? AB - The metabolism of haloperidol (HP) to the potentially neurotoxic pyridinium species, HPP+ and RHPP+, has been demonstrated in humans. In vitro studies in microsomes harvested from various animal species indicate that the tetrahydropyridines, HPTP and RHPTP, could be intermediates in this pathway. However, this has not yet been demonstrated in vivo in humans. In this study, plasma and urine collected from eight critically ill patients treated with high doses of intravenous HP were analyzed for HPTP and RHPTP using HPLC with electrochemical detection. However, neither HPTP nor RHPTP were detected despite plasma concentrations of HP and RHP higher than any previously reported. HPP+ and RHPP+ were both present in the urine in high concentrations and accounted for 1.1 +/- 0.5% and 5.3 +/- 3.6%, respectively, of the administered dose of HP. The apparent elimination half-lives of HPP+ and RHPP+ were 67.3 +/- 11.0 hr and 63.3 +/- 11.6 hr, respectively. The absence of HPTP and RHPTP in plasma and urine suggests that in humans these tetrahydropyridines either are insignificant intermediates in the metabolism of HP in vivo or are present only transiently at their site of formation and are not released into the circulation. PMID- 9399631 TI - Hormonal regulation of intestinal 11beta-hydroxysteroid dehydrogenase. AB - We have previously demonstrated the developmental increase of the activity of 11beta-hydroxysteroid dehydrogenase (11betaHSD) in the rat ileum which correlated with the developmental surge of plasma concentrations of corticosteroids, thyroid hormones and insulin. To ascertain whether these hormones directly stimulate 11betaHSD activity we used explant cultures of ileum and distal colon. The intestinal segments of young, 7-day-old rats, were cultured 48 hours in the presence of aldosterone (10[-7] M), dexamethasone (10[-7] M), triiodothyronine (10[-7] M) or insulin (10[-7] M) and 11betaHSD activity was evaluated by measuring the conversion of [3H]corticosterone to [3H]11-dehydrocorticosterone. The activity of 11betaHSD was significantly increased following 48 h treatment with dexamethasone and aldosterone, whereas insulin and triiodothyronine were without any effect. Corticosterone oxidation was inhibited by carbenoxolone and progesterone. It is being concluded, that both glucocorticoids and mineralocorticoids but not insulin or triiodothyronine induce intestinal 11betaHSD activity. PMID- 9399632 TI - TGF-beta2 prevents the impaired chondrocyte proliferation induced by unloading in growth plates of young rats. AB - Growth plate width and cartilage organization are altered during skeletal unloading in growing rats. Immunohistochemical studies have identified TGF-beta in calcified cartilage, and TGF-beta is known to induce mitogenic effects on chondrocytes in vitro. On the other hand, IGF-1 was shown to be expressed in the proximal tibial growth plate and to mediate GH-induced longitudinal bone growth in rats. We therefore investigated the effect of recombinant human (rh) IGF-1 and rhTGF-beta2 infusion on the changes induced by unloading in the cellular organization of the growth plate in growing rats. Hindlimb unloading for 14 days induced a 13% reduction in growth cartilage height in the proximal tibia. This effect was mostly related to a 17% and 14% decrease in the proliferative zone height and chondrocyte number, respectively. In unloaded rats treated with a systemic infusion of rhTGF-beta2 (2microg/kg/day) the number of chondrocytes in the proliferative zone was not different from those of normal loaded animals. In contrast, rhIGF-1 treatment at a 2mg/kg/day dose was not effective in counteracting the effects of unloading on growth plate height and chondrocyte number. These results show that systemic administration of rhTGF-beta2 prevents in large part the reduced growth of chondrocytes in the proliferative zone and the reduced epiphyseal growth plate growth induced by unloading in rats. PMID- 9399633 TI - Reduced kidney branched chain aminotransferase expression in puromycin aminonucleoside-induced nephrotic syndrome. AB - Injection of puromycin aminonucleoside to rats induces nephrotic syndrome characterized by hypoalbuminemia, proteinuria and hypercholesterolemia. In these rats, a low protein diet (6% casein diet) increased serum albumin by 26.3%, decreased proteinuria by 39% and reduced total cholesterol by 32%. Branched chain aminotransferase activity in kidney mitochondria of nephrotic rats fed 20 or 6% casein diet decreased by 30 and 24% with respect to their pair-fed groups and it was not modified by the protein content of the diet. Mitochondrial branched chain aminotransferase mRNA expression decreased by 67.3 and 72.5% in nephrotic rats fed 20 and 6% casein diet in comparison to their pair-fed groups. Total serum branched chain amino acids concentration (leucine, isoleucine, valine) in nephrotic rats was 30% higher than their pair-fed groups and it was associated with a decrease in the branched chain aminotransferase activity and mRNA expression suggesting that the catabolism of branched chain amino acid is reduced to conserve body nitrogen. PMID- 9399634 TI - The transcription of the XRCC1 gene in spleen following ionizing irradiation in radiosensitive and radioresistant mice. AB - The XRCC1 gene was described to play a role for the sensitivity of mammalian cell lines towards ionizing irradiation. Cells with a mutation of this gene present with decreased single strand break repair, reduced recombination repair, show increased double strand breaks and sister chromatid exchange is increased up to tenfold. The goal of our study was to investigate the transcription of this gene in the spleen following ionizing irradiation in the mouse. Furthermore, we intended to examine whether radiation sensitive (RS) mice would show a transcriptional pattern different from radiation resistant (RR) mice. Radiation sensitive BALB/c/J Him mice and radiation resistant C3H He/Him mice were untreated or whole body irradiated with X-ray at 4 and 6 Gy and sacrificed 5, 15 and 30 min after irradiation. mRNA was isolated from the spleen and hybridized with probes for XRCC1 and beta-actin as a house keeping gene control. Transcription of XRCC1 was not different in unirradiated or 4 Gy-irradiated mouse RR or RS mouse strains. When irradiated at 6 Gy, RR mice showed an approximately threefold increase of mRNA XRCC1/mRNA beta actin as early as 15 min after irradiation. We conclude that radiation resistant mice show a higher transcription level for the XRCC1 gene in the spleen early after high dose X-ray whole body irradiation. This finding is the first in vivo study on XRCC1 of this kind and may in part explain the differences in the radiation sensitivity between the two strains studied. PMID- 9399635 TI - Isoflurane anesthesia blunts cerebral responses to noxious and innocuous stimuli: a fMRI study. AB - We used functional magnetic resonance imaging to determine how isoflurane affected cerebral neuronal activation resulting from noxious and innocuous stimuli. Five male volunteers were subjected to mild electrical shock and tactile stimuli applied to the hand. During low (0.7%) and moderate (1.3%) isoflurane anesthesia the stimuli were repeated and a supramaximal electrical shock was also applied. Tactile stimulation activated bilateral SI and SII, but resulted in no significant activation at low or moderate anesthesia. Electrical shock activated contralateral SI and bilateral SII; low anesthesia completely abolished this response. The supramaximal stimulus activated the caudate nucleus and bilateral thalamus at low anesthesia; these responses were diminished at moderate anesthesia. Isoflurane anesthesia blunts cerebral responses to somatosensory stimuli, and the absence of cortical activation during supramaximal stimulation suggests that noxious-induced movement is generated in lower CNS structures. PMID- 9399636 TI - Chronic steroid sulfatase inhibition by (p-O-sulfamoyl)-N-tetradecanoyl tyramine increases dehydroepiandrosterone sulfate in whole brain. AB - Dehydroepiandrosterone sulfate (DHEAS) is a neurosteroid which functions as a negative allosteric modulator of the GABA(A) receptor-gated chloride channel. Steroid sulfatase inhibitors including (p-O-sulfamoyl)-N-tetradecanoyl tyramine (DU-14), can potentiate the blockade of the amnestic effects of scopolamine by exogenously administered DHEAS. Moreover, when administered over a 15 day period, DU-14 can block scopolamine amnesia without the concurrent administration of DHEAS. Since the enzyme, steroid sulfatase, facilitates the hydrolysis of the sulfate moiety from DHEAS, the intent of this study was to determine whether chronic administration of DU-14 could increase whole brain concentrations of endogenous DHEAS. Rats were administered DU-14 or corn oil vehicle for 15 days. Following the last day the animals were sacrificed and the brains were removed and analyzed for DHEAS content. DU-14 increased the whole brain concentration of DHEAS 77.6%, from 0.65 +/- 0.06 to 1.15 +/- 0.12 microg/g (mean +/- SEM). This result suggests that steroid sulfatase inhibitors may enhance cognitive function following chronic treatment by increasing the concentration of excitatory neurosteroids such as DHEAS in the brain. Steroid sulfatase inhibitors, therefore, may provide a novel mechanism for facilitating central nervous system function. PMID- 9399637 TI - Effect of dopamine on immune cell proliferation in mice. AB - Dopamine is known as a precursor of catecholamine and one of the neurotransmitters in brain and peripheral tissues. Recent studies suggest an important role of dopamine in immune responses. In the present study, intraperitoneal administration of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) which lowered endogenous dopamine suppressed splenocyte proliferation in response to mitogens such as lipopolysaccharide (LPS) and concanavalin A (Con A). Moreover, intravenous injection of the specific agonists of dopamine DA-1 receptor (SKF38393) or DA-2 receptor (LY171555) into mice enhanced the splenocyte proliferation stimulated by LPS or Con A. In the in vitro cultures, dopamine, SKF38393 and LY171555 directly promoted cell proliferation to LPS or Con A. These results indicate that dopamine has an ability to regulate B- and T-cell proliferation both in vivo and in vitro. PMID- 9399638 TI - Aspalatone, a new antiplatelet agent, attenuates the neurotoxicity induced by kainic acid in the rat. AB - The antioxidant efficacy of aspalatone (APT; acetyl salicylic acid maltol ester), a new antiplatelet agent, has been characterized in vivo as well as in vitro, and several observations indicated that the antioxidant could prevent the neuroexcitation caused by oxidative stress. In this report, the effect of APT was evaluated on kainic acid (KA)-induced neurotoxicity, since the neurotoxicity induced by KA is, at least in part, mediated via the formation of free radicals. The results showed that pretreatments with APT or maltol (MAL) significantly attenuated seizure activity, oxidative stress (lipid peroxidation and protein oxidation) and the loss of hippocampal neurons induced by KA. On the other hand, the pretreatments with aspirin (ASP), ASP together with MAL or vitamin E failed to protect against the toxicity produced by KA suggesting that the mechanism of action for APT on the KA-induced neurotoxicity is different from that of ASP. These finding raise the possibility that salicylmaltol, a metabolite of APT, plays a role in preventing the neurotoxicity evoked by KA. Therefore, our results suggest that an APT-related antioxidant mechanism, which is linked to the MAL moiety, is involved in the neuroprotective effect against KA. PMID- 9399639 TI - Cbl-b, a member of the Sli-1/c-Cbl protein family, inhibits Vav-mediated c-Jun N terminal kinase activation. AB - We have used the yeast two-hybrid system to identify proteins that interact with Vav, a GDP/GTP exchange factor for the Rac-1 GTPase that plays an important role in cell signaling and oncogenic transformation. This experimental approach resulted in the isolation of Cbl-b, a signal transduction molecule highly related to the mammalian c-cbl proto-oncogene product and to the C. elegans Sli-1 protein, a negative regulator of the EGF-receptor-like Let23 protein. The interaction between Vav and Cbl-b requires the entire SH3-SH2-SH3 carboxy terminal domain of Vav and a long stretch of proline-rich sequences present in the central region of Cbl-b. Stimulation of quiescent rodent fibroblasts with either epidermal or platelet-derived growth factors induces an increased affinity of Vav for Cbl-b and results in the subsequent formation of a Vav-dependent trimeric complex with the ligand-stimulated tyrosine kinase receptors. During this process, Vav, but not Cbl-b, becomes highly phosphorylated on tyrosine residues. Overexpression of Cbl-b inhibits the signal transduction pathway of Vav that leads to the stimulation of c-Jun N-terminal kinase. By contrast, expression of truncated Cbl-b proteins and of missense mutants analogous to those found in inactive Sli-1 proteins have no detectable effect on Vav activity. These results indicate that Vav and Cbl-b act coordinately in the first steps of tyrosine protein kinase receptor-mediated signaling and suggest that members of the Sli 1/Cbl family are also negative regulators of signal transduction in mammalian cells. PMID- 9399640 TI - Oncogenic Ras-induced secretion of a novel inhibitor of skeletal myoblast differentiation. AB - Expression of oncogenic H-Ras in 23A2 myoblasts (A2:H-Ras cells) is sufficient to induce both a transformed phenotype and a differentiation-defective phenotype. Because oncogenic Ras is known to induce the secretion of several different growth factors involved in maintaining the transformed phenotype of both fibroblast and epithelial cells, we explored the possibility that expression of oncogenic Ras in 23A2 myoblasts might lead to the secretion of a factor which inhibits differentiation. The differentiation of 23A2 myoblasts was inhibited (i) by coculture with an equal number of A2:H-Ras cells, (ii) by culture with an equal number of A2:H-Ras cells in the same tissue culture medium on an insert which allowed equilibration of molecules smaller than 1 micron, and (iii) by culture in media previously conditioned by A2:H-Ras cells. Similar results were obtained when 23A2 myoblasts expressing oncogenic N-Ras were substituted for A2:H Ras cells in each assay. No inhibition of differentiation was observed, however, when differentiation-defective E1A-expressing 23A2 cells or C3H10T1/2 fibroblasts were substituted for A2:H-Ras cells. The differentiation inhibitor(s) in media conditioned by A2:H-Ras cells is heat stable, larger than 3 kD, and sensitive to the non-specific growth factor antagonist, suramin. Western analyses failed to detect either FGF-2 or TGFbeta (the known inhibitors of myoblast differentiation) in media conditioned by A2:H-Ras cells. Furthermore, while FGF-2 is a potent activator of MAP kinase and TGFbeta is a potent inhibitor of mink lung epithelial cell (CCL64) growth, conditioned media from A2:H-Ras cells does not activate MAP kinase and does not inhibit the growth of CCL64 cells. These results indicate that expression of oncogenic Ras induces the secretion of a novel inhibitor of skeletal myoblast differentiation. Furthermore, these results are the first to implicate an autocrine/paracrine mechanism in the inhibition of differentiation by oncogenic Ras. PMID- 9399641 TI - Insulin activates Stat3 independently of p21ras-ERK and PI-3K signal transduction. AB - The binding of insulin to its receptor initiates multiple signal transduction pathways regulating such diverse processes as proliferation, differentiation, glucose transport, and glycogen metabolism. The STAT-family of transcription factors has been demonstrated to play a critical role in gene induction by a variety of hemopoietic cytokines and hormones. Furthermore, constitutive activation of STATs is observed in transformed cells. Here we describe activation of a transcriptional complex binding to a consensus STAT-transcriptional element in response to insulin challenge. This complex is induced rapidly after tyrosine autophosphorylation of the insulin receptor, and is sustained for several hours. Supershift analysis of the insulin-induced complex reveals that it specifically contains the transcription factor Stat3. DAN binding of this complex is inhibited by pre-incubation with tyrosine, but not serine/threonine protein kinase inhibitors, whereas transcriptional activation is inhibited by both. Utilising a dominant negative mutant of p21ras we demonstrate that both insulin-induced Stat3 DNA-binding and also transactivation do not require p21ras. Furthermore, although previous studies have suggested a role for MAP kinases (ERKs) and PI-3K in STAT activation, utilising the specific MEK inhibitor PD098059 and the PI-3K inhibitor wortmannin, we demonstrate that activation of ERKs or PI-3K are not required for insulin induced Stat3 phosphorylation or transactivation. PMID- 9399642 TI - Biological activity of p27kip1 and its amino- and carboxy-terminal domains in G2/M transition of Xenopus oocytes. AB - p27kip1 is a general inhibitor of Cdks that preferentially accumulates and functions during G1 phase, before the restriction point of the mammalian cell cycle. We observed that injection of purified p27kip1 into Xenopus oocytes potently inhibits the G2/M transition and activation/dephosphorylation of the maturation promoting factor (MPF, p34cdc2/cyclin B complex) kinase associated with germinal vesicle breakdown (GVBD) induced by progesterone or insulin. Addition of exogenous p27kip1 in vitro to lysates of hormonally matured oocytes blocked the enzymatic activity of the activated MPF kinase present in those extracts. Interestingly, the isolated amino-terminal region of p27kip1 (p27N), encompassing only the Cdk binding site, exhibited a similar inhibitory behavior in vitro and a weaker inhibitory effect in vivo than the complete p27kip1 protein. Surprisingly, the remaining carboxy-terminal region of p27kip1 (p27C) actually induced GVBD when injected alone into the oocytes, and also accelerated the kinetics of insulin- or progesterone-induced GVBD. Consistent with the in vivo observations, p27C formed a complex with, and activated, the MPF kinase in lysates of immature oocytes, although this activation was blocked by simultaneous addition of p27N or complete p27kip1. Active MPF was able to phosphorylate p27C only in the absence of p27N or whole p27kip1, suggesting that the inhibitory activity associated with the amino terminus is dominant over the activation produced by p27C. These results demonstrate the functional interaction of p27kip1 with cyclin B/p34cdc2 complexes during G2/M progression in oocytes, and suggest that the amino and carboxy terminal portions of this protein may play opposite regulatory roles, reminiscent of the corresponding N- and C-terminal portions of p21waf. We speculate that accumulation of a truncated, C-terminal p27 fragment may play a physiological regulatory role in progression through G2 and later stages of the cell cycle. PMID- 9399643 TI - SCH 51344-induced reversal of RAS-transformation is accompanied by the specific inhibition of the RAS and RAC-dependent cell morphology pathway. AB - RAS interacts with multiple targets in the cell and controls at least two signaling pathways, one regulating extracellular signal-regulated kinase (ERK) activation and the other controlling membrane ruffling formation. These two pathways appear to act synergistically to cause transformation. SCH 51344 is a pyrazolo-quinoline derivative identified based on its ability to derepress transformation sensitive alpha-actin promoter in RAS-transformed cells. Previous studies have shown that SCH 51344 is a potent inhibitor of RAS-transformation. However, SCH 51344 had very little effect on the activities of proteins in the ERK pathway, suggesting that it inhibits RAS-transformation by a novel mechanism. In this study, we show that SCH 51344 specifically blocks membrane ruffling induced by activated forms of H-RAS, K-RAS, N-RAS and RAC. Treatment of fibroblast cells with this compound had very little effect on RAS-mediated activation of ERK and JUN kinase activities. SCH 51344 was effective in inhibiting the anchorage-independent growth of Rat-2 fibroblast cells transformed by the three forms of oncogenic RAS and RAC V12. These results indicate that SCH 51344 inhibits a critical component of the membrane ruffling pathway downstream from RAC and suggest that targeting this pathway may be an effective approach to inhibit transformation by RAS and other oncogenes. PMID- 9399644 TI - Cdk2-dependent phosphorylation of p27 facilitates its Myc-induced release from cyclin E/cdk2 complexes. AB - Activation of Myc triggers a rapid induction of cyclin E/cdk2 kinase activity and degradation of p27. Overt degradation of p27 is preceded by a specific dissociation of p27 from cyclin E/cdk2, but not from cyclin D/cdk4 complexes. We now show that cyclin E/cdk2 phosphorylates p27 at a carboxy-terminal threonine residue (T187) in vitro; mutation of this residue to valine stabilises cyclin E/cdk2 complexes. This reaction is not significantly inhibited by high concentrations of p27, suggesting that cdk2 bound to p27 is catalytically active. In vivo, p27 bound to cyclins E and A, but not to D-type cyclins is phosphorylated. Myc-induced release of p27 from cdk2 requires cdk2 kinase activity and is delayed in a T187V mutant of p27. After induction of Myc, p27 phosphorylated at threonine 187 transiently accumulates in a non cdk2 bound form. Our data suggest a mechanism in which p27 is released from cyclin E/cdk2 upon phosphorylation; in Myc-transformed cells, release is efficient as phosphorylated p27 is transiently bound in a non-cdk2 containing complex and subsequently degraded. PMID- 9399645 TI - Characterization of human N8 protein. AB - We have shown before that the N8 mRNA is expressed at higher levels in lung tumor and lung tumor-derived cell lines than normal lung cells. In this paper, we have characterized the N8 protein, and studied its properties. The N8 gene encodes a major 24 kDa protein and its expression correlates well with the N8 mRNA expression pattern observed in different cell lines. N8 protein is capable of forming a homodimer or multimeter in vitro. It is a phosphorylated cytoplasmic protein and phosphorylation occurs mainly at serine residues. N8 protein is expressed at higher levels in epithelial cells than in mesenchymal cells. N8 protein expression is induced in a fibroblast cell line expressing adenoviral Ela protein, which acquired epithelial-like characteristics. Furthermore, ectopic expression of N8 protein in NIH3T3 cells converts them into a spheroid form. These spheroids also have some of the characteristic features of epithelial cells. Taken together, these results suggest that the N8 protein may be associated with the development or maintenance of epithelial cell phenotype. PMID- 9399646 TI - Reexpression of the retinoblastoma protein in tumor cells induces senescence and telomerase inhibition. AB - Normal human diploid cells senesce in vitro and in vivo after a limited number of cell divisions. This process known as cellular senescence is an underlying cause of aging and a critical barrier for development of human cancers. We demonstrate here that reexpression of functional pRB alone in RB/p53-defective tumor cells via a modified tetracycline-regulated gene expression system resulted in a stable growth arrest at the G0/G1 phase of the cell cycle, preventing tumor cells from entering S phase in response to a variety of mitogenic stimuli. These cells displayed multiple morphological changes consistent with cellular senescence and expressed a senescence-associated beta-galactosidase biomarker. Further studies indicated that telomerase activity, which was assumably essential for an extended proliferative life-span of neoplastic cells, was abrogated or repressed in the tumor cell lines after induction of pRB (but not p53) expression. Strikingly, when returned to an non-permissive medium for pRB expression, the pRB-induced senescent tumor cells resumed DNA synthesis, attempted to divide but most died in the process, a phenomenon similar to postsenescent crisis of SV40 T-antigen transformed human diploid fibroblasts in late passage. These observations provide direct evidence that overexpression of pRB alone in RB/p53-defective tumor cells is sufficient to reverse their immortality and cause a phenotype that is, by all generally accepted criteria, indistinguishable from replicative senescence. The results suggest that pRB may play a causal role in the intrinsic cellular senescence program. PMID- 9399647 TI - Role of wild type p53 in the G2 phase: regulation of the gamma-irradiation induced delay and DNA repair. AB - Upregulation of the p53 protein was shown to induce cell cycle arrest at the G1/S border and in some cases at the G2/M border. Furthermore, it was suggested that p53 is associated with the induction of the various DNA repair pathways. Previously, we demonstrated that cells co-expressing endogenous wild type p53 protein, together with dominant negative mutant p53, exhibit deregulation of apoptosis, G1 arrest and delay in G2 following gamma-irradiation. In the present study, we investigated the role of p53 protein in the DNA damage response at the G2 phase. Using p53-null, wild type p53 and mutant p53-producer cell lines, we found that the two C-terminally spliced p53 forms could prevent gamma-irradiation induced mutagenesis prior to mitosis, at the G2/M checkpoint. We found that at the G2 phase, p53 may facilitate repair of DNA breaks giving rise to micronuclei, and regulate the exit from the G2 checkpoint. At the G1 phase, only the regularly spliced form of p53 caused growth arrest. In contrast, both the regularly and the alternatively spliced p53 forms directed postmitotic micronucleated cells towards apoptosis. These results provide a functional explanation for the cell cycle independent expression of p53 in normal cycling cells, as well as in cells where p53 is up-regulated, following DNA damage. PMID- 9399648 TI - Methylation of the multi tumor suppressor gene-2 (MTS2, CDKN1, p15INK4B) in childhood acute lymphoblastic leukemia. AB - The multi tumor suppressor genes MTS1 (CDKN2 p16INK4A) and MTS2 (CDKN1, p15INK4B) located at 9p21-22 are inactivated in some human cancers via several mechanisms including deletion and hypermethylation. We have investigated the deletion and methylation status of MTS1 and MTS2 in childhood acute lymphoblastic leukemia (ALL) of both T-cell (17 cases) and B-cell phenotypes (29 cases), and p16INK4A and p15INK4B mRNA expression in 36 of these cases. Biallelic or monoallelic loss of both MTS1 and MTS2 was observed in 12 cases of B-ALL and nine cases of T-ALL. Two cases of T-ALL showed deletion of MTS1 but not MTS2. The 5' CpG region of MTS2 was hypermethylated in 12 cases of precursor B-ALL and eight cases of T-ALL but no hypermethylation was found in the 5' CpG region of MTS1. All cases with homozygous deletion of MTS1 or MTS2 had no or low levels of mRNA expression and similar low levels of expression were found in cases in which MTS2 was present but fully methylated. Thus hypermethylation of MTS2, in contrast to MTS1, is frequent in childhood ALL. Furthermore our data show that although inactivation of MTS1 by deletion is common, inactivation of MTS2 by a combination of deletion and hypermethylation is more frequent in both B-ALL (20/29, 69%) and T-ALL (17/17, 100%). This suggests that both MTS1 and MTS2 are important targets of the 9p21-22 deletion. PMID- 9399649 TI - Malignant transformation by cyclin E and Ha-Ras correlates with lower sensitivity towards induction of cell death but requires functional Myc and CDK4. AB - We demonstrate in this paper that the G1 phase specific cell cycle regulator cyclin E is able to provoke focus formation when cotransfected with activated Ha ras into primary rat embryo fibroblasts (REFs). Cyclin E/Ha-ras transformed cells are highly tumorigenic in synergeneic rats, are able to form colonies in soft agar and show protection towards apoptosis upon serum starvation or DNA damage compared to cells transformed by the combination of Myc, cyclin D1 or SV40 large T-antigen and Ha-ras. Lines that were established after cyclin E/Ha-ras or cyclin D1/Ha-ras transformation contain a large percentage of polyploid cells. This was not observed in cells transformed with other oncoproteins and Ha-ras pointing to an involvement of D- and E type cyclins in genomic instability. The cyclin dependent kinase inhibitors p21 and p27 but also p16 completely abrogate focus formation by cyclin E and Ha-ras suggesting that the oncogenic activity of cyclin E still requires functional G1 specific cyclin/CDK complexes. Moreover, inhibition of Myc function also blocks the oncogenic activity of cyclin E indicating a requirement of Myc for cyclin E function. The findings presented here demonstrate that cyclin E can act as an oncoprotein with a potential involvement in genomic instability and the prevention of cell death. Our data also present more evidence for a strict functional interdependency between G1 cyclin/CDK complexes and c-Myc. PMID- 9399650 TI - Suppression of morphological transformation by radicicol is accompanied by enhanced gelsolin expression. AB - Radicicol, an inhibitor of Src-family protein-tyrosine kinases, causes morphological reversion of v-src- and v-Ha-ras-transformed fibroblasts and arrest of the cell cycle at both the G1 and the G2 phases. Radicicol was found to inhibit the growth of several other oncogene-transformed cell lines and human carcinoma cell lines and to revert their cell morphology to be flat. In the radicicol-treated flat cells, actin stress fiber bundles were reorganized. Since this effect of radicicol on these cell lines was inhibited by cycloheximide, de novo protein synthesis is required for the morphological reversion. Screening of cellular proteins enhanced in response to radicicol by two-dimensional gel electrophoresis suggested that the amount of gelsolin, an actin regulatory protein, was distinctly increased upon radicicol treatment. Western blot and Northern blot analyses showed that radicicol enhanced transcription of the gelsolin gene in human carcinoma cell lines, as a result of which the amount of gelsolin was increased several folds. Injection with an anti-gelsolin antibody into cells and successive treatment with radicicol resulted in approximately 80% reduction of the number of flat cells with stress fibers in comparison with controls treated with an irrelevant antibody. These results show that elevated expression of gelsolin is associated, at least in part, with the suppression of transformation and the restoration of actin stress fibers in human carcinoma cells by radicicol. PMID- 9399652 TI - Epstein-Barr-virus-associated gastric cancer in Russia. AB - The present investigation was carried out to estimate the prevalence of EBV associated cases among gastric carcinoma (GC) patients of Russia and the Republics of the former Soviet Union (FSU). With this aim, formalin-fixed paraffin-embedded blocks from 206 gastric carcinomas obtained from patients of the Cancer Research Center, Moscow, were investigated by EBV-encoded RNA-1 (EBER 1) in situ hybridization applied to paraffin sections. As a result, 18 GC cases (8.7%) revealed uniform EBER-1 expression restricted to the carcinoma cells. Hybridized signals not detected in non-neoplastic gastric epithelium. EBV involvement was significantly more frequent among males, especially in tumors of less differentiated types (moderately differentiated tubular adenocarcinomas and poorly differentiated solid adenocarcinomas) and located in the upper stomach (cardia and middle). Most EBV-positive GCs were characterized by strong lymphoid compartment involvement. Our findings concerning the distribution of EBV-positive GCs by sex, site and hystological type are similar to those in Japan, however, EBV-positive rate of GC cases in Russia is higher than in Japan and lower than in USA. PMID- 9399651 TI - A sero-epidemiological study of the relationship between sexually transmitted agents and cervical cancer in Honduras. AB - To investigate a possible cause-and-effect relationship between sexually transmitted diseases and cervical cancer, we performed a sero-epidemiological study on the presence of antibodies against a number of sexually transmitted agents (STAs) in patients with cervical cancer and their matched controls. In this study, we used serological techniques to investigate the presence of antibodies to cytomegalovirus, herpes simplex virus type 2, human immunodeficiency virus, Chlamydia trachomatis, Treponema pallidum and human papillomavirus (HPV) early protein E7 in sera from patients with cervical cancer, cervical intra-epithelial neoplasia and individually matched, healthy controls. The presence of antibodies to infectious agents other than HPV appeared not to be associated with risk of cervical neoplasia in either univariate or multivariate analysis. After adjustment for cytology, schooling and presence of HPV DNA in cervical scrapes, there was a significantly higher prevalence of antibodies to HPV-16 E7 protein in sera from patients with cervical cancer (OR = 3.6, 95% CI 1.0-12.9) than in healthy controls. The highest antibody prevalence was found among HPV-16 DNA-positive cervical cancer patients (33%). Our results indicate that in these study groups past infections with the STA considered seems to be of no apparent relevance for cervical carcinogenesis and that the HPV-16 anti-E7 response appears to be associated with cervical cancer. PMID- 9399653 TI - Induction of a T- and B-cell response against a unique amino acid sequence of the mouse IgG2A hinge region in a MAb-treated patient. AB - A patient with malignant glioblastoma was treated with intratumoral infusions of the murine MAb425 (IgG2A) directed against the epidermal growth factor receptor. At the 10th infusion, the patient developed somnolence, fever and headache. The symptoms increased during the subsequent 48 hr but then gradually disappeared within a week. The cerebrospinal fluid (CSF) contained increased concentrations of interleukin-2. The main CSF cell subset was CD4 T-cells. A marked blood lymphocyte proliferative response against mouse IgG2A was noted. The reactive T cell epitope(s) could be localized to a 14 amino acid (RGPTIKPCPPCKCP) long peptide of the hinge region. A B-cell response (IgG antibodies) against this peptide was also induced. PMID- 9399654 TI - A novel gastric-cancer-associated mucin antigen defined by a monoclonal antibody A3D4. AB - De-glycosylation of mucins may expose new tumor-associated core protein epitopes. In this study, to attempt to develop useful markers for gastric cancers, we have purified and de-glycosylated gastric mucin and tried to establish monoclonal antibodies (MAbs). A MAb designated A3D4 among established MAbs was shown to react with gastric cancer with high frequency, but not with normal gastric epithelium. Among normal digestive organs, only the colon and gall bladder were positive for MAb A3D4. The incidence of positivity in gastric cancer was 75% for intestinal-type adenocarcinoma (n = 28), 40% for solid-type adenocarcinoma (n = 5) and 33% for signet/scirrhous-type adenocarcinoma (n = 15). Interestingly, adenoma and intestinal metaplasia (IM) with chronic gastritis or peptic ulcer were negative for MAb A3D4, whereas 8 out of 13 cases (62%) of IM with gastric cancer was positive. Western-blot analysis using the lysate from normal colon tissues revealed a high-molecular-weight (> 300-kDa) smear-like band. Immunohistochemical analysis indicated that the reactivity of MAb A3D4 was clearly increased when tissue sections were pre-treated with periodic acid or O glycanase, while it was decreased by pre-treatment with trypsin or protease V8. There was no reactivity with the synthetic peptide encompassing the tandem-repeat sequence of MUC2 or MUC3. These data suggest that MAb A3D4 detects a novel gastric-cancer-associated mucin antigen whose epitope may be peptide in nature. PMID- 9399655 TI - p53 mutations in childhood thyroid tumours from Belarus and in thyroid tumours without radiation history. AB - Mutations in the p53 tumour-suppressor gene (exons 5-8) were investigated in 31 Belarussian childhood thyroid tumours (24 cases of papillary thyroid carcinoma, 3 benign tumours and 2 cases each of thyroiditis and goiter); 33 thyroid tumours from juveniles and adults without radiation exposures (25 carcinomas of various histological types, including 11 papillary carcinomas and 8 adenomas) and 6 tumours from adults (4 papillary carcinomas, 1 adenoma, 1 goiter) served as controls. The mutational spectrum of p53 differed greatly between the childhood thyroid carcinomas from Belarus and the control groups. In the control groups of 29 malignant thyroid tumours, 7 different mutations were detected on exons 5-8, none of which occurred among the 15 papillary carcinomas in this group. Five mutations were found in tissue samples of the 24 childhood papillary carcinomas, and they were all the same p53 point mutation (CGA --> CGG) on codon 213 of exon 6. To determine whether this mutation is simply a polymorphism or whether it is specific to the tumour cells, laser-assisted microdissection was applied to collect various areas of tumorous and non-tumorous cells (10-20 cells per sample) from each paraffin-embedded tissue section of 8 of the papillary thyroid carcinomas. Using PCR-SSCP and sequence analysis on these cells, the very same p53 mutation on codon 213 was detected in various microdissected tumour samples of 2 cases, but it was not found in any microdissected non-tumorous sample. The exclusive occurrence of this p53 mutation in selective microdissected samples of tumour cells, even as homozygous mutation in 1 case, reflects a distinct tumour heterogeneity within papillary childhood thyroid carcinomas. PMID- 9399656 TI - Incidence of prostatic intra-epithelial neoplasia in Osaka, Japan. AB - High-grade prostatic intra-epithelial neoplasia (HGPIN) is the most likely precancerous lesion for prostatic carcinoma. A high incidence of its association with cancer has been reported in Western countries. On the other hand, information regarding its incidence is limited in Japan, where the mortality due to prostate cancer is much lower. We reviewed 53 clinical stage T2 or T3 prostatic cancers of Japanese patients living in Osaka, Japan (mean age, 67.2 years). These cases were subdivided into a pre-operatively non-castrated group (34 cases) and a medically or surgically castrated group (19 cases). HGPIN was found in 27 cases. The incidence of HGPIN was significantly lower in the castrated group (21.0%) compared with the non-castrated group (67.6%). In the non castrated group, patient age, pathological stage, Gleason score, tumor size and serum prostate-specific antigen showed no significant correlation with HGPIN. Advanced pathological stage and tumor size tended to decrease the incidence of HGPIN, although this was not statistically significant. When the study group was limited to stage T2 tumors of the non-castrated group, the incidence of HGPIN was 81.0%. HGPIN in Japan may also be clinically and etiologically significant as a precursor of clinical cancer. PMID- 9399657 TI - Calcitonin receptors, bone sialoprotein and osteopontin are expressed in primary breast cancers. AB - Several human breast cancer cell lines express the calcitonin receptor (CTR), but this has not been demonstrated previously in clinical breast cancers. We examined 18 primary breast cancers by reverse transcription-PCR, for expression of CTR and of the bone proteins osteopontin (OPN) and bone sialoprotein (BSP). OPN and CTR were expressed by each of the tumours, and 7 (39%) additionally expressed an alternate form of CTR, whilst BSP was expressed by 13 tumours (72%). In situ hybridisation confirmed that expression of OPN and CTR was confined to the tumour cells. Expression of CTR, BSP and OPN may prove to be a useful marker for breast cancers, and their role in the homing of breast cancer cells to bone remains to be investigated. PMID- 9399658 TI - Heterogeneous p53 mutations in a Burkitt lymphoma from an AIDS patient with monoclonal c-myc and VDJ rearrangements. AB - This study investigates the timing of p53 mutations detected in the malignant cells of a Burkitt's lymphoma cell line (BRG-P) with respect to other maturation or transforming events. The BRG-P cell line, derived from an AIDS patient, was of special value since it displayed subclones that had undergone an isotype switch from IgM to IgA1 (BRG-M and BRG-A cells). BRG-M and BRG-A cells were characterized by the same monoclonal c-myc and VDJ rearrangements and by the expression of Ig receptors with specificity for a 45 kDa protein of human breast cells. Analysis of p53 mutations in the different BRG subclones showed that 1) BRG-M cells displayed 2 different p53 mutations in trans; since the original BL cells also showed the same mutations, this finding indicated that both occurred in vivo; 2) one of the p53 alleles of BRG-A cells was lost, while the other showed a mutation different from those seen in BRG-M cells; and 3) all 3 mutations observed in BRG-M or BRG-A cells resulted in the functional inactivation of the transcriptional activation function of p53. Together, our data demonstrate that p53 mutations were relatively late events during lymphomagenesis. Moreover, in view of the role of p53 in cell apoptosis, it is conceivable that BRG cells were subjected to a strong selective pressure that favored p53 inactivation. Such inactivation was possibly required to counterbalance other potentially apoptotic events, including the presence of a deregulated c-myc oncogene and signals delivered by the host environment in situ. PMID- 9399659 TI - Cancer risk in first-degree relatives of children with malignant tumours (Province of Trieste, Italy). AB - We conducted a population-based cohort study in the province of Trieste, Italy, to assess whether the first-degree relatives of children with malignancies had an increased risk of cancer compared with the general population. We examined cancers occurring in all first-degree relatives of children who experienced malignancies under the age of 15 years between 1971 and 1993 (probands). A cohort of the 394 relatives of the 125 probands contributed 7,939 person-years of observation. Among the relatives as a whole, we found a statistically significant increased risk of developing all malignancies except non-melanoma skin carcinoma (21 observed relatives with cancer and 12.46 expected, for a standardized incidence ratio [SIR] of 1.69), of developing breast cancer (SIR = 3.09) and of developing haemolymphatic system neoplasms (SIR = 4.03). This was mainly due to the excess cancer risk in the relatives of probands with intracranial tumours, who showed a significant 3.1-fold risk for developing all cancers but non melanoma skin tumours. Our findings and the previously reported steep rise in the incidence of childhood brain tumours in our area may imply that not only genetic factors but also shared environmental agents might be involved in the observed aggregation of cancer in the families of probands with intracranial tumours. PMID- 9399660 TI - Occupational exposure to polyvinyl chloride as a risk factor for testicular cancer evaluated in a case-control study. AB - Occupational exposures were assessed in a case-control study on testicular cancer using self-administered questionnaires. In total, answers were obtained for 148 (91%) cases and 315 (87%) controls. Of the cases, 101 had seminoma and 47 had embryonal testicular cancer. An increased odds ratio (OR) was found for exposure to polyvinyl chloride (PVC) yielding an OR of 6.6 (95% confidence interval, 1.4 32). The risk increased further if cases with self-reported cryptorchidism or orchitis were excluded. Six of the 7 exposed cases had seminoma. Exposure to other types of plastics did not significantly increase the risk of testicular cancer. PMID- 9399663 TI - Risk of second cancers in classical Kaposi's sarcoma. AB - An association between Kaposi's sarcoma (KS) and malignant lymphoma has been suspected for many years. Both cancers belong to the group of malignancies associated with immune suppression and have been known to occur in the same individual. Accordingly, a common etiology has been suspected. Through linkage within the Nordic cancer registries, we studied the occurrence of cancers in a population-based cohort of 741 patients with classical KS. The relative risk of subsequent malignancies was expressed as the ratio of the observed numbers of cancer to the numbers expected based on age-, sex-, period- and country-specific incidence rates, i.e., the standardized incidence ratio (SIR). A total of 104 cancers was observed during 5,802 person-years of follow-up, which was close to the expected 98.8 cases (SIR, 1.05). During the first year of follow-up, 3 lymphomas were observed, which is in significant excess of the 0.2 lymphomas expected (SIR, 13.0). In contrast, no lymphomas occurred in the period beyond the first year of follow-up vs. 2.3 expected. Cancers of the buccal cavity and pharynx (SIR, 10.6; n = 4) and of the colon (SIR, 2.7; n = 7) occurred in excess among women but not among men. Accordingly, our results indicate that patients with classical KS are not at increased risk of cancer in general. In particular, the overall risk of lymphomas was not significantly increased. The high relative risk of malignant lymphoma immediately after KS was based on a limited number of cases, and this observation is unlikely to indicate a common etiology. PMID- 9399661 TI - Germline hMSH2 and hMLH1 gene mutations in incomplete HNPCC families. AB - Hereditary non-polyposis colon cancer (HNPCC) is a common hereditary disease characterized by a predisposition to an early onset of colorectal cancer. The majority of the HNPCC families carry germline mutations of either hMSH2 or hMLH1 genes, whereas germline mutations of hPMS1 and hPMS2 genes have rarely been observed. Almost all of the germline mutations reported so far concern typical HNPCC families. However, there are families that display aggregations of colon cancer even though they do not fulfil all HNPCC criteria (incomplete HNPCC families) as well as sporadic cases of early onset colon cancers that could be related to germline mutations of these genes. Therefore, we screened germline mutations of hMSH2 and hMLH1 genes in 3 groups of patients from France and Turkey: typical HNPCC (n = 3), incomplete HNPCC (n = 9) and young patients without apparent familial history (n = 7). By in vitro synthesis of protein assay, heteroduplex analysis and direct genomic sequencing, we identified 1 family with hMSH2 mutation and 5 families with hMLH1 mutations. Two of the 3 HNPCC families (66%) displayed hMLH1 germline mutations. Interestingly, 4 of 9 families with incomplete HNPCC (44%) also displayed mutations of hMSH2 or hMLH1 genes. In contrast, no germline mutation of these genes was found in 7 young patients. Our results show that germline mutations of hMSH2 and hMLH1 genes contribute to a significant fraction of familial predisposition to colon cancer cases that do not fulfil all diagnostic criteria of HNPCC. PMID- 9399662 TI - Histological diagnosis of Helicobacter pylori gastritis is predictive of a high risk of gastric carcinoma. AB - Chronic Helicobacter pylori infection has been identified as a major risk factor for the subsequent development of gastric carcinoma On the basis of seroepidemiological studies the relative risk for infected persons was estimated to range between 3 and 6. Our study attempted to determine the relative risk of gastric carcinoma in H. pylori-infected individuals based on the histological evaluation of gastritis in gastric carcinoma patients in the light of a declining prevalence of H. pylori infection in Western countries. We histologically determined the H. pylori infection rate in 215 patients with early gastric carcinoma (tumor stage pT1), and compared it with that of 215 asymptomatic persons matched by age and sex who were tested by the 13C urea breath test. On the basis of these data an odds ratio of 16.7 (CI 9.6-29.1) was calculated for the relative risk of developing gastric carcinoma in H. pylori-infected people. The histological diagnosis of gastritis permits a separate risk assessment for patients with autoimmune gastritis, and by excluding these patients from the analysis we calculated an odds ratio for H. pylori-infected persons of 150 (CI 36.4-622.9). The endoscopic-histological diagnosis of H. pylori infection is associated with an increased risk of the subsequent development of gastric carcinoma of approximately 150-fold compared with H. pylori-negative patients who do not have chronic atrophic corpus gastritis of the autoimmune type (type A gastritis). PMID- 9399664 TI - Genetic immunotherapy by intrapleural, intraperitoneal and subcutaneous injection of IL-2 gene-modified Lewis lung carcinoma cells. AB - The induction and augmentation of tumor non-specific immunity and of tumor specific immunity by intrapleural, intraperitoneal and subcutaneous injection of interleukin-2 (IL-2) gene-modified Lewis lung carcinoma (LLC) cells (LLC-IL2) was tested in C57BL/6 mice. Intrapleural injection of LLC cells induced lung tumors with a malignant effusion, intraperitoneal injection induced peritoneal tumors with ascites and subcutaneous injection induced subcutaneous tumors. Intrapleural injection of irradiated LLC-IL2 cured pre-existing lung LLC tumors and extended the survival of the mice but did not affect survival of mice with pre-existing peritoneal tumors nor did it affect the growth of s.c. tumors. Intraperitoneal injection of irradiated LLC-IL2 cured pre-existing LLC peritoneal tumors and extended the survival of the mice but did not affect survival of mice bearing lung tumors nor did it affect the growth of s.c. tumors. Subcutaneous injection of irradiated LLC-IL2 did not affect the growth of preexisting s.c. tumors and also did not improve survival of mice bearing the lung or peritoneal tumors. Injection with irradiated LLC-IL2 by all routes, i.e., intrapleural, intraperitoneal and s.c., protected against subsequent re-challenge with LLC. Eight days after the initial immunization (early stage of immunization), non adherent mononuclear cells in the peritoneal cavity of the mice treated with intraperitoneal injection of irradiated LLC-IL2 displayed enhanced cytotoxicity against LLC, B16-F10 and P815 cells, while the cytotoxic activity of spleen cells in the same mice did not change. The efficiency of induction of tumor-specific immunity was the strongest after intraperitoneal immunization and weakest after s.c. immunization. In vitro analysis using the spleen cells of mice immunized with irradiated LLC-IL2 suggested that CD8+ T cells play a key role in tumor specific immunity. PMID- 9399665 TI - Remodeling of glycoconjugates on CD44 enhances cell adhesion to hyaluronate, tumor growth and metastasis in B16 melanoma cells expressing beta1,4-N acetylglucosaminyltransferase III. AB - Beta1-4 N-acetylglucosaminyltransferase III (GnT-III) synthesizes bisecting N acetylglucosamine structures on asparagine-linked oligosaccharides. Using B16-hm mouse melanoma cells stably expressing GnT-III activity as positive transfectants, the effect of bisecting N-acetylglucosamine on the function of CD44 was analyzed in association with adhesion to hyaluronate and tumor spread in mice. Transfection of GnT-III caused increased affinity of immunoprecipitated CD44 to erythro-agglutinating phytohemagglutinin, that preferentially recognizes bisecting N-acetylglucosamine, without affecting the surface CD44 amount, indicating an increase in bisecting N-acetylglucosamine residues on CD44 in positive transfectants. CD44-mediated adhesion to immobilized hyaluronate and the binding of fluorescence-labeled hyaluronate to the cell surface were increased in positive transfectants. The enhanced adhesion in positive transfectants was suppressed by the treatment with beta-N-acetylhexosaminidase, indicating that N acetylglucosamine residues were responsible for the enhanced adhesion. Positive transfectants showed promoted CD44-mediated tumor growth and metastatic development in the spleen after subcutaneous inoculation into mice. These results indicate that glycosylation of CD44 due to GnT-III causes enhanced adhesion to hyaluronate, local tumor growth and metastatic growth in spleen, suggesting that the CD44-mediated adhesion and tumor spread can be modified through introduction of a glycosyltransferase gene. PMID- 9399666 TI - Folate-maytansinoids: target-selective drugs of low molecular weight. AB - Folate receptor is over-expressed in a variety of carcinomas. To design a cytotoxic drug that would selectively target these carcinomas, we synthesized folate-maytansinoids. These drugs showed high affinity toward folate receptor, appeared to enter cells exclusively via the folate receptor-mediated caveolar pathway and displayed high cytotoxic potency (in the range of 10[-11] to 10[-10] M) and remarkable selectivity for folate receptor-expressing carcinoma cell lines. Folate-maytansinoids represent a new class of tumor-specific agents in which the targeting and the cytotoxic function can be altered independently. PMID- 9399668 TI - Neutral metoclopramide sensitizes cytotoxicity induced by ionizing radiation in SCID mice xenografted with a human brain astrocytoma. AB - A formulation of metoclopramide (MCA) conformationally altered by neutralization of pH (nMCA, Neu-Sensamide) has been shown to have the same efficacy of enhancing the cytotoxicity of a single dose of 1 Gy radiation as acidic formulations (e.g., Primperan, Sensamide) in a human lung adenocarcinoma (H2981) xenografted into SCID mice. In the present study, 2 x 1 Gy radiation was combined with 2 x 2 mg nMCA/kg body weight injected 2 hr before radiation treatment for evaluation of radiosensitization in SCID mice xenografted with a human brain astrocytoma (T24). Given in this treatment schedule, nMCA alone at 2 mg/kg showed no cytotoxic effect on tumor growth in vivo. When combined with 2 x 1 Gy of radiation, however, the cytotoxicity was significantly increased as measured by tumor growth delay over the radiation-only-treated group. Furthermore, nMCA was absorbed into brains of mice and rats as efficiently as acidic MCA (aMCA) when analyzed 45 min after i.m. injection by high-performance liquid chromatography. PMID- 9399667 TI - Targeting the tumor vasculature: inhibition of tumor growth by a vascular endothelial growth factor-toxin conjugate. AB - Tumor-derived vascular endothelial growth factor (VEGF)/ vascular permeability factor (VPF) plays an important role in neovascularization and the development of tumor stroma. Furthermore, VEGF receptors are over-expressed in the endothelial cells of tumor vasculature and almost non-detectable in the vascular endothelium of adjoining normal tissues. The differential expression of receptor offers a selective advantage for targeting cytotoxic toxin polypeptides. We have prepared a vascular targeting reagent by chemically linking recombinant VEGF to a truncated form of diphtheria toxin. The VEGF-toxin conjugate was selectively toxic to endothelial cell lines and inhibited experimental neovascularization of the chick chorioallantoic membrane. In the present study, we examined the effects of VEGF-toxin conjugate on solid tumor growth. Athymic nude mice with established subcutaneous tumors were treated with daily intraperitoneal injections of the VEGF-toxin conjugate or free toxin. When compared with control animals treated with the toxin polypeptide alone, the conjugate-treated animals displayed a significant inhibition of tumor growth. Histological analysis of tumors from conjugate-treated animals revealed hemorrhagic necrosis consistent with a vascular-mediated injury. In contrast, highly vascularized normal tissues from conjugate-treated animals demonstrated no evidence of hemorrhage or tissue injury. The conjugate was well tolerated without apparent toxicities. Our results illustrate the anti-tumor activity of a VEGF-toxin conjugate selectively targeting the tumor neovasculature. PMID- 9399669 TI - Soluble HER-2/neu neutralizes biologic effects of anti-HER-2/neu antibody on breast cancer cells in vitro. AB - Amplification and over-expression of the HER-2/neu proto-oncogene are associated with poor prognosis in women with both node-positive and node-negative breast cancer. Therefore, the encoded surface glycoprotein represents an attractive target for cancer immunotherapies. Furthermore, the extracellular domain of HER 2/neu is released from the cell surface by proteolytic cleavage. In the present experiments, we investigated the potential biologic effects of soluble HER-2/neu with particular emphasis on its interaction with anti-HER-2/neu antibodies. A monoclonal antibody specific for the extracellular domain of HER-2/neu dose dependently inhibited the proliferation of highly HER-2/neu-expressing SK-BR-3 and BT-474 breast cancer cells but had no effect on the proliferation of weakly to moderately HER-2/neu-expressing MCF-7, HBL-100 and ZR-75-1 breast cells. Addition of SK-BR-3 or BT-474 cell supernatants with high concentrations of soluble HER-2/neu led to a neutralization of anti-HER-2/neu antibody-mediated inhibition of proliferation due to a binding of soluble HER-2/neu by the antibody, which could be demonstrated by immunoprecipitation. Furthermore, the ability of anti-HER-2/neu antibodies to mediate antibody-dependent cellular cytotoxicity (ADCC) by lymphokine-activated killer cells was assessed. Cytolysis of SK-BR-3 tumor cells was increased significantly in the presence of anti-HER 2/neu antibodies. Similar to the proliferation inhibition, ADCC was neutralized by addition of soluble HER-2/neu-containing supernatants. Our data suggest that tumors rich in HER-2/neu might thus escape certain steps of immunologic control by neutralizing biologic activities of anti HER-2/neu antibodies due to the presence of soluble HER-2/neu. PMID- 9399670 TI - Effect of aspirin on cell proliferation and differentiation of colon adenocarcinoma Caco-2 cells. AB - Several lines of evidence suggest that long-term treatment with non-steroidal anti-inflammatory drugs may reduce the risk of colon cancer and the size and number of colonic polyps in patients with familial adenomatous polyposis. Aspirin has also been shown to inhibit cell proliferation in human tumor cell lines and to induce apoptosis in colonic mucosa of familial polyposis patients. To elucidate the molecular mechanisms of the antiproliferative action of aspirin, we studied the effects of aspirin on cell growth and differentiation of the human colon carcinoma Caco-2 cell line. These cells represent a useful tool for studying the mechanisms involved in the regulation of cell growth and differentiation of intestinal epithelial cells since they spontaneously differentiate into polarized cells, expressing brush border enzymes. We show in this study that aspirin (0.1-10 mM) induces a profound inhibition of cell replication as assessed either by cell counts or thymidine incorporation. Moreover, aspirin concentrations of 5 and 10 mM induce apoptosis, whereas concentrations of 1 and 2 mM do not. The inhibition of growth is associated with a dose-dependent reduction in insulin-like growth factor II mRNA expression and with an increase in sucrase activity (a brush border enzyme) and apolipoprotein A I mRNA expression, 2 specific markers of the differentiative status of this cell line. Our data thus show that aspirin-dependent inhibition of cell growth is associated with the enterocyte-like differentiation of Caco-2 cells. PMID- 9399671 TI - Paracrine interactions between mesothelial and colon-carcinoma cells in a rat model. AB - This study used a co-culture system with Transwell tissue-culture inserts to investigate the role of primary cultures of rat peritoneal mesothelial cells on the proliferation of rat colon-carcinoma cells (CC531 cells). Mesothelial cells significantly inhibited the growth of CC531 cells, while, conversely, CC531 cells stimulated the growth of mesothelial cells. Receptor-binding studies demonstrated the presence of high-affinity IGF-I receptors on the mesothelial and CC531 cells. Both cell types also produced IGF-I, as measured by radioimmunoassay. IGF-I stimulated DNA synthesis in mesothelial cells, but had no effect on the growth of CC531 cells. In co-culture, it was found that IGF-I potentiated the inhibitory effect of mesothelial cells on CC531 cells. The effect of IGF-I on mesothelial cell proliferation was additive to the stimulatory effect of CC531 cells. TGF beta had no effect on the growth of the CC531 cells, suggesting that this growth (-inhibitory) factor is not involved in the inhibitory effect of mesothelial cells on CC531 cell growth. The study provides evidence for the existence of a paracrine loop between mesothelial and colon-carcinoma cells, giving more insight into the basic cellular mechanisms that may modulate the growth of intraperitoneal colon carcinoma. Inhibition of CC531-cell proliferation by rat mesothelial cells might explain the earlier finding that tumour cells grow poorly in a surgically uncompromised abdomen. PMID- 9399672 TI - Anti-tumor activity of CPT-11 in experimental human ovarian cancer and human soft tissue sarcoma. AB - CPT-11, a semi-synthetic derivative of camptothecin, was investigated for its activity in panels of 15 human ovarian-cancer lines and 10 human soft-tissue sarcoma lines grown s.c. in nude mice. Various factors were analyzed that may be of influence on the extent of tumor-growth inhibition induced by CPT-11. At equitoxic doses, CPT-11 was more effective in the daily x5 schedule than the weekly x2 schedule, although a 2-fold higher dose was administered in the weekly x2 schedule. Since i.p. and i.v. injections were similarly effective, the selected treatment schedule was 20 mg/kg i.p. daily x5, starting when tumors measured approximately 150 mm3. Growth inhibition of > or = 75% was obtained in 8/15 human ovarian-cancer lines and in 6/10 human soft-tissue sarcoma lines. A weak correlation was found between topoisomerase-I mRNA in xenograft tissues and sensitivity to CPT-11. Relative topoisomerase-I expression was highest in MRI-H 207 and WLS-160 xenografts, in which CPT-11 was able to induce cures of all tumors. The high efficacy in the 2 panels of human tumor lines suggests over prediction of its potential clinical activity in these tumor types. The difference in efficacy of CPT-11 between species may be related to the metabolism of the drug, since CPT-11 is converted more efficiently into SN-38 in mice. In addition, mice may be less sensitive to SN-38-induced side-effects. On the basis of the preclinical data, frequent administration of lower doses of CPT-11 should be considered in order to increase response rates in the clinic. PMID- 9399673 TI - Induction of glutathione S-transferase pi as a bioassay for the evaluation of potency of inhibitors of benzo(a)pyrene-induced cancer in a murine model. AB - There is a growing need for short-term and cost-effective bioassay to assess the efficacy of potential chemo-preventive agents. We report that the induction of glutathione (GSH) S-transferase pi (mGSTP1-1) by a chemo-preventive agent can be used as a reliable marker to assess its efficacy in retarding chemical carcinogenesis induced by benzo(a)pyrene (BP), which is a widespread environmental pollutant and believed to be a risk factor in human chemical carcinogenesis. This conclusion is based on 1) the relative contribution of mGSTP1-1 of the liver and forestomach of female A/J mice in the detoxification of the ultimate carcinogenic metabolite of BP, (+)-anti-7,8-dihydroxy-9, 10-oxy 7,8,9, 10-tetrahydrobenzo(a)pyrene [(+)-anti-BPDE]; and 2) a positive correlation between the induction of hepatic and forestomach mGSTP1-1 by 5 naturally occurring organosulfides (OSCs) from garlic (diallyl sulfide, diallyl disulfide, diallyl trisulfide, dipropyl sulfide and dipropyl disulfide) and their effectiveness in preventing BP-induced forestomach neoplasia in mice. In the liver, the combined contribution of other GSTs in the detoxification of (+)-anti BPDE was far less than the contribution of mGSTP1-1 alone. Likewise, in the forestomach, the contribution of mGSTP1-1 far exceeded the combined contribution of other GSTs. Studies on the effects of OSCs against BP-induced forestomach neoplasia revealed a good correlation between their chemo-preventive efficacy and their ability to induce mGSTP1-1 expression in the liver (r = -0.89; p < 0.05) as well as in the forestomach (r = -0.97; p < 0.05). Our results suggest that the induction of mGSTP1-1 may be a reliable marker for evaluating the efficacy of potential inhibitors of BP-induced cancer in a murine model. PMID- 9399674 TI - Le(y) glycolipid acts as a co-factor for tumor procoagulant activity. AB - We have generated a monoclonal antibody (MAb), FS01, which inhibits the procoagulant activity (CCA-1) produced by a human squamous cell carcinoma cell line, LK52. Expression of the antigen recognized by FS01 MAb in various cancer cell lines correlated well with the procoagulant activities of the expressing cell lines. Our objective was to characterize the molecule reacting with FS01 MAb and to analyze its involvement in the CCA-1 procoagulant activity. The molecule was identified as a glycolipid and found to be involved in the procoagulant activity because both procoagulant activity and reactivity to FS01 MAb were lost after endoglycoceramidase treatment of CCA-1. Furthermore, FS01 MAb recognized the Lewis Y (Le[y]) antigen. To confirm the involvement of a glycolipid incorporating the Le(y) antigen in the procoagulant activity, we attempted to purify CCA-1 from LK52 culture supernatant. In one of the purification steps, a fraction containing low procoagulant activity (CCA-1p) separated from the Le(y) positive fraction (CCA-1c). Although CCA-1c alone did not show procoagulant activity, the procoagulant activity of CCA-1p was augmented by CCA-1c and this augmentation was inhibited by FS01 MAb. Furthermore, CCA-1c enhanced the procoagulant activity of 33 cell lines tested as well as CCA-1p. In addition, purified Le(y) glycolipid from canine intestine augmented the procoagulant activity of CCA-1p, and this augmentation also could be inhibited by FS01 MAb. We conclude that Le(y) glycolipid is a co-factor for the procoagulant activity derived from cancer cells. PMID- 9399675 TI - Androgens are not a direct requirement for the proliferation of human prostatic epithelium in vitro. AB - The androgen receptor pathway is known to be a key regulator of growth in the normal and pathological prostate. However, the precise mechanisms of this signaling pathway with respect to the different cellular compartments of the prostate remain largely unknown. We have used a primary culture system to grow human prostatic epithelial cells of normal, benign, tumor and metastatic origin, as well as immortalized human prostatic epithelial cell lines, to demonstrate the absence of a direct or indirect effect of androgens on cellular proliferation in vitro. In parallel to this observed androgen independence for growth, all cell systems lost significant expression of androgen receptor, prostate-specific antigen and prostatic acid phosphatase. Since the androgen receptor is expressed in the epithelium in situ, our results suggest that the androgen effect on epithelial cells may be one of prostatic differentiation rather than proliferation, and that the androgen receptor/growth factor pathway acts through mesenchymal-epithelial interactions. PMID- 9399676 TI - Role of heparin-binding EGF-related peptides in proliferation and apoptosis of activated ras-stimulated intestinal epithelial cells. AB - The ras mutation is a common and critical step in carcinogenesis. Autocrine growth factors are also known to play an important role in cancer cell growth and transformation. However, the contribution of autocrine growth factors in regulation of proliferation and apoptosis of activated ras-stimulated intestinal epithelium is not fully understood. Therefore, we constructed activated ras transfected intestinal epithelial cell clones (IEC-ras) to examine the role of epidermal growth factor (EGF)-related peptides in the behavior of IEC-ras. Overexpression of EGF family growth factors (transforming growth factor alpha, heparin-binding EGF-like growth factor, amphiregulin and betacellulin) and stronger phosphorylation of the EGF receptor was observed in IEC-ras compared with control cells. IEC-ras proliferated more rapidly than control cells, and a specific EGF receptor kinase inhibitor, AG 1478, abolished the increased proliferation of IEC-ras. Heparitinase and chlorate also prevented increased proliferation of IEC-ras. Additionally, IEC-ras expressed more bcl-2 and was more resistant to apoptosis induction by UV radiation and mitomycin C. AG 1478 suppressed bcl-2 expression and inhibited resistance to apoptosis of IEC-ras. Heparitinase and chlorate had effects similar to those of AG 1478. Our data indicate that heparin-binding EGF family growth factors play an important role in both increased proliferation and resistance to apoptosis of ras-stimulated intestinal epithelial cells. PMID- 9399677 TI - 5-Fluorouracil-resistant colonic tumors are highly responsive to sodium butyrate/interleukin-2 bitherapy in rats. AB - Advanced colorectal cancer is generally refractory to 5-fluorouracil (5-FU) chemotherapy. This is linked to the emergence of resistant cell populations, probably due to a selection process. The identification of molecular markers and the improvement of alternative therapies thus remain important. We have used as an experimental model a rat colon cancer cell line (PROb), which exhibits features similar to those of the human situation. 5-FU treatment of rats bearing PROb tumors enhanced their survival but did not lead to cure. A PROb 5-FU resistant subline (PRObR1) was obtained by continuous in vitro exposure to 5-FU. Resistance to 5-FU was accompanied by a 2-fold increase in thymidylate synthase activity and a substantially higher incorporation of thymidine in the presence of 5-FU, compared with parental PROb cells. Unexpectedly, in syngeneic rats, PRObR1 tumors exhibited delayed growth when compared with parental PROb tumors. This was ascribed to an increased sensitivity of PRObR1 cells to host immune response since no growth delay was observed in immunocompromised nude mice and since there was no detectable difference in proliferation rates between PROb and PRObR1 cells. 5-FU treatment was inefficient in prolonging the survival of rats bearing PRObR1 tumors. In contrast, an immunotherapeutic protocol combining sodium butyrate and recombinant interleukin-2 (NaBut/rIL-2) cured 80% of the rats bearing established PRObR1 tumors. Our results suggest that NaBut/rIL-2 treatment is efficient against 5-FU-chemoresistant rat colon cancer. PMID- 9399679 TI - Focusing in on the medial temporal lobe. PMID- 9399680 TI - Puzzles of psychiatric genetics--new candidate gene approaches. PMID- 9399681 TI - Depressing transmission in GABAergic hippocampal neurons. PMID- 9399682 TI - The search for infectious agents in neuropsychiatric disorders: lessons from multiple sclerosis. PMID- 9399683 TI - Mood disorders across the continents. PMID- 9399684 TI - Computational functions of the hippocampus: does it encode all episodic memories? PMID- 9399685 TI - Serotonin transporter mRNA in schizophrenia. PMID- 9399686 TI - Interactions among genes for transcription factors in hypothalamic neurons: implications for reproductive behaviors. AB - Gene products for nuclear hormone receptors, which are transcription factors, interact in the cell nuclei of neurons in a manner important for the hypothalamic control of instinctive behaviors. In doing so, the combinatorial logic of their competition or synergy may serve to integrate environmental and physiological constraints upon those behaviors. PMID- 9399687 TI - Changes in brain gene expression in psychiatric illness: mRNA differential display provides some clues. AB - The 20th century has witnessed a progressive increase in our understanding of brain structure, organisation and function and now includes knowledge at the macromolecular and ionic levels. Investigations of such diverse functions as cognition, memory and mood, performed mainly in whole animal studies, are now advancing rapidly with the application of modern molecular biological techniques. In this article we consider the contribution of mRNA differential display to the analysis of altered gene expression in vitro and in vivo. The role that this technique may play in the identification of genes involved in the aetiology of psychiatric disorders and their treatment is discussed. PMID- 9399688 TI - Serotonin transporter gene polymorphisms: ethnic difference and possible association with bipolar affective disorder. AB - There is some evidence suggesting that a polymorphism of variable number of tandem repeats (VNTR) in the second intron of the serotonin transporter (5-HTT) gene and another variation which lies 1.2 kb upstream of the promoter of the gene (5-HTTLPR) are associated with affective disorders. However, conflicting results have also been reported. We examined an allelic association of these two polymorphisms in a Japanese sample of 191 patients with affective disorders (142 bipolar and 49 unipolar) and 212 controls. Substantial differences in the number and frequency of alleles between Caucasians and Japanese were observed for both polymorphisms. A significant association between the VNTR polymorphism and bipolar disorder (genotypic association: odds ratio 2.2, 95% CI 1.2-4.0; allelic association: odds ratio 1.7, 95% CI 1.0-3.0) was found, but not between the 5 HTTLPR polymorphism and bipolar disorder. No significant association with unipolar depression was detected using either genetic marker, although this may be attributable to the relatively small number of subjects with unipolar depression. Our results suggest that the VNTR itself or another unknown functional polymorphism which would be in linkage disequilibrium to the VNTR has an effect on susceptibility to bipolar disorder. PMID- 9399690 TI - Additional evidence for an association between the dopamine D4 receptor (D4DR) exon III repeat polymorphism and the human personality trait of Novelty Seeking. AB - The long alleles (> or =6 repeats) of the dopamine D4 dopamine receptor exon III polymorphism have been linked in some, but not all, studies to Novelty Seeking (NS), one of four personality traits defined by Cloninger's tridimensional personality questionnarie (TPQ). In order to further examine the robustness of our original observation we have recruited an additional cohort similar in demographic structure to the original cohort. Although no significant difference in mean NS scores was observed when the new subjects (n=94) were grouped by presence (NS=17.83 +/- 1.16) or absence (NS=16.45 +/- 0.65) of the 7 repeat allele, a significant difference in range of NS scores was observed (non parametric Moses range test, P = 0.01). The effect of the seven allele was also significant in those individuals scoring highest on NS (>1 standard deviation from the mean; t-test, t=5.13, P=0.002). In the expanded cohort (n=218) a significant effect of the seven allele on NS is demonstrated by both parametric (t=2.26, P=0.01) and non-parametric (range test, P=0.004) statistical tests. The effect is also observed in both principal ethnic groups (Ashkenazi and non Ashkenazi Jews). In the expanded cohort the effect is significant in female (t=2.2, P=0.03, n=98) but not in male subjects (t=1.12, P>0.1, n=116). We discuss both direct and indirect evidence that in our opinion continues to support a modest role for the long alleles of the dopamine D4 receptor repeat polymorphism in the determination of NS behavior at least in some population groups. PMID- 9399689 TI - Cerebral benzodiazepine receptor binding and distribution in generalized anxiety disorder: a fractal analysis. AB - Data obtained from animal and human brain imaging studies indicate that frontal cortex and medial temporal lobe are involved in experiencing and controlling fear and anxiety. We tested the hypothesis that benzodiazepine receptor binding is decreased in the left temporal pole and increased in the right prefrontal area among patients suffering from anxiety. We studied 10 drug-naive female patients with generalized anxiety disorder (GAD) and 10 age- and gender-matched healthy controls with MRI and with SPET by using a new (123)I-labelled specific benzodiazepine receptor radioligand, NNC 13-8241. Blindly analyzed results showed that the benzodiazepine receptor binding of [(123)I]NNC 13-8241 was significantly decreased in the left temporal pole among patients with GAD when compared with age- and sex-matched healthy controls. This hemispheric asymmetry was studied further with a fractal analysis of the SPET images. The fractal dimension of the left hemispheric benzodiazepine receptor binding in patients with GAD was significantly higher than that of controls (1.28 +/- 0.09 and 1.17 +/- 0.07, respectively), whereas the intercept was decreased by 43 +/- 23% reflecting more homogeneous cerebral benzodiazepine receptor density distribution in patients with GAD. The finding is analogous to the decreased heterogeneity of myocardial blood flow observed in patients with ischemic heart disease. The results are consistent with the general hypothesis that high regional heterogeneity of perfusion, metabolism and receptor density is necessary to maintain adaptation ability in the living organism. PMID- 9399692 TI - Lack of linkage between the corticotropin-releasing hormone (CRH) gene and bipolar affective disorder. AB - Corticotropin-releasing hormone (CRH) plays a key role in the regulation of the stress response. Abnormalities in CRH secretion have been documented in both the depression and manic phases of bipolar disorder (BPD). In the present study, we investigated genetic linkage between the CRH gene and BPD in 22 pedigrees. A highly informative, short tandem repeat (STR) polymorphism adjacent to the CRH gene on human chromosomal region 8q13 was used to examine linkage. Affected sibling pair (ASP) and the likelihood-based disequilibrium tests revealed nonsignificant values. We conclude that the CRH gene is not linked to BPD; if genes involved in the regulation of stress response are indeed linked to BPD, the search should be directed towards those that regulate CRH secretion or its effects on target tissues. PMID- 9399691 TI - No evidence for expanded polyglutamine sequences in bipolar disorder and schizophrenia. AB - Several recent studies have suggested that expanded CAG repeats may contribute to the genetic transmission of bipolar disorder and schizophrenia. In all known disorders associated with expanded CAG repeats, the repeat sequence is translated into glutamine. Therefore the simplest hypothesis is that one or more proteins with expanded polyglutamine sequences are involved in the pathogenesis of bipolar disorder and schizophrenia. In order to examine this hypothesis, we have used an antibody against expanded polyglutamine sequences to examine Western blots prepared from lymphoblastoid cell lines of patients with schizophrenia and bipolar disorder. We also examined Western blots prepared from left frontal cortex tissue samples obtained from 11 schizophrenics post mortem. With the exception of the TATA-binding protein (TBP), we did not detect any proteins containing expanded polyglutamine sequences. Our data therefore suggest either that the expanded repeats which are associated with these disorders do not encode polyglutamine, or that they are within genes that are not expressed within the tissues investigated here. PMID- 9399693 TI - Genotypic selection provides experimental confirmation for an alcohol consumption quantitative trait locus in mouse. AB - Quantitative genetic research has produced a wealth of basic information concerning genetic influence on alcohol-related processes. Recent developments in quantitative trait locus (QTL) methodology were promptly applied to the task of individuating polygenes affecting alcohol-related attributes in animal models and a body of reliable data is gradually coming into focus as a result of replication and convergence of evidence from a variety of methods. A key issue in QTL research is the need to distinguish true positive results from the false positive results that are inherent in analytical procedures requiring large numbers of significance tests. One school of thought holds that stringent significance levels should be imposed; another suggests more modest criteria for QTL nomination, with subsequent confirmation trials with independent samples. Recombinant inbred strains and various types of intercrosses have been used in correlational designs, both for nomination and confirmation studies. Alternative experimental procedures include knockout preparations and short-term phenotypic selective breeding. We present here results from a third experimental method-that of marker-based genotypic selection--in evaluation of two nominated QTLs for alcohol acceptance in mice. PMID- 9399694 TI - Mu opioid receptor gene variants: lack of association with alcohol dependence. AB - The mu opioid receptor is implicated in the reward, tolerance and withdrawal effects of alcohol and other drugs of abuse. This hypothesis is supported by the effects of alcohol on beta-endorphin release, of mu opioid receptor agonists and antagonists on alcohol consumption, and by the activation of the dopaminergic reward system by both alcohol and opiates. In addition, the murine mu opioid receptor locus, Oprm, is implicated as the major quantitative trait locus (QTL) affecting the different levels of morphine consumption between two inbred mouse strains that also exhibit differences in alcohol and cocaine consumption. Detection of genetic variation affecting OPRM1 expression or mu opioid receptor function would be an important step towards understanding the origins of inter individual variation in response to mu opioid receptor ligands and in diseases of substance dependence. We directly sequenced the human mu opioid receptor locus, OPRM1, to detect natural variation that might affect function and/or be associated with psychiatric phenotypes related to opioid function. Four DNA sequence variants were found: three non-synonymous substitutions (Ala6Val [rare], Asn40Asp, [0.10-0.16], Ser147Cys [rare]) and one intronic variant (IVS2+691G/C [0.55-0.63]). OPRM1 alleles, genotypes and haplotypes from three psychiatrically characterized population samples (US Caucasian [USC, n=100], Finnish Caucasian [FC, n=324] and Southwestern American Indian [SAI, n=367]), were used to perform association and sib-pair linkage analyses with alcohol and drug dependence diagnoses. No significant association of OPRM1 genetic variation to phenotype was observed. This analysis has 80% power to detect a small to moderate effect of OPRM1 variation on alcohol dependence and 100% power to detect effects of the magnitude of the ALDH2*2 variant. While these data do not support a role of the mu opioid receptor in susceptibility to alcohol dependence, the potential relationship between OPRM1 genetic variation and response to endogenous opioids and exogenous opiates can now be investigated. PMID- 9399695 TI - Prodynorphin mRNA expression is increased in the patch vs matrix compartment of the caudate nucleus in suicide subjects. AB - Experimental and clinical studies suggest an involvement of the opioid neuropeptide system in psychiatric disorders. Notably, opioid peptide immunoreactivity is altered in the cerebrospinal fluid of chronic schizophrenics and manic-depressive subjects. Despite these clinical findings, few postmortem investigations have examined the association of endogenous opioid neuropeptides with schizophrenia and suicide. Anatomically, a tight interaction exists within the neostriatum between the opioid peptide (dynorphin and enkephalin) system and classical neurotransmitters such as dopamine which has been implicated in both the psychotic symptoms and the cognitive deficits that characterize schizophrenia (see review). The neostriatum is differentially organized into patch and matrix neurochemical mosaic compartments anatomically connected to limbic- and sensorimotor-related brain regions, respectively. Moreover, the human neostriatum is characterized by a heterogenous expression of the prodynorphin opioid gene: high in the patch, but low in the matrix compartment. The present results show for the first time a differential alteration of prodynorphin within distinct striatal compartments in postmortem tissue from nonschizophrenic suicide subjects. The prodynorphin patch/matrix mRNA expression was elevated in the caudate nucleus of suicide subjects as compared to normal controls and schizophrenics in which no alterations in opioid peptides or D1 and D2 mRNA expression were apparent. Altogether the findings suggest that discrete dysfunction of the endogenous opioid dynorphin system might contribute to depression and the risk of suicide in nonschizophrenic subjects. PMID- 9399696 TI - An integrated physical map of 18p11.2: a susceptibility region for bipolar disorder. AB - The reported linkage between bipolar disorder and a large pericentric portion of chromosome 18 has been replicated in an independent study. Further examination of this region showed that 18p11.2 had the greatest allele sharing in our pedigrees and increased sharing in other independently ascertained pedigree series permitting refinement of the region of significance. To facilitate positional cloning of a susceptibility gene, we used a combination of mapping reagents, including a subchromosomal somatic cell hybrid panel, a contig of clones in yeast artificial chromosomes (YAC), and a radiation hybrid (RH) panel, to construct a high resolution physical map of the region including sequence tag sites (STSs) and expressed sequence tags (ESTs). This approach generated the interlocus distance and order of 15 STSs and 16 ESTs including four novel transcripts, with an average of approximately 200 kb between loci, over a approximately 6-Mb region. This high resolution integrated map will be an important tool in providing loci for contig construction, and positional candidates for mutation screening. PMID- 9399697 TI - Cerebral changes and cerebrospinal fluid beta-amyloid in Alzheimer's disease: a study with quantitative magnetic resonance imaging. AB - Pathological and biochemical studies indicate that beta-amyloid (betaA4) deposition is a hallmark in the pathogenesis of Alzheimer's disease (AD). Neuroimaging studies demonstrate that the respective cerebral changes primarily strike the temporal lobe and the amygdala-hippocampus complex and may be reliably assessed using quantitative magnetic resonance imaging (MRI). Therefore one may expect that reduced betaA4-levels are significantly correlated with measures of the temporal lobe rather than global cerebral atrophy in AD patients. To test this hypothesis in a clinical study, cerebrospinal fluid concentrations of total betaA4 and its major C-terminal variations betaA4 1-40 and betaA4 1-42 were compared with cerebral changes as assessed by quantitative magnetic resonance imaging (MRI). Significantly (P< 0.05) reduced betaA4 1-40 and betaA4 1-42 levels were found in the AD patients (17 female; six male; AD/NINCDS-ADRDA-criteria) in comparison to the patients with major depression (seven female; two male; DSM-III R). Within the AD group, betaA4 and betaA4 1-42 levels were significantly correlated with the volume of the temporal lobes (r= 0.46 and r= 0.48, respectively) but none of the other volumetric measures. These findings indicate that changes in cerebral betaA4 levels contribute to temporal lobe atrophy in AD and support the possibility that betaA4 is central to the etiology of AD. PMID- 9399698 TI - Transcriptional regulation of the human IL5 gene by ionizing radiation in Jurkat T cells: evidence for repression by an NF-AT-like element. AB - Eosinophilia is often observed in patients with parasitic infections and atopic diseases like allergic asthma and atopic dermatitis. Additionally, it is a typical feature of the inflammatory reaction after therapeutic and accidental exposure to ionizing radiation. This uniquely specific phenomenon regulated by the cytokine interleukin 5 (IL-5) suggests specific control for IL5 gene expression. In this study, we generated promoter-CAT constructs containing different human IL-5 promoter regions spanning from positions -507 to +43. Transfection experiments in Jurkat T cells revealed that the promoter sequence from -57 to +43 was required for constitutive and inducible IL-5 promoter activity. Low baseline CAT activity could be enhanced by treatment with phenylmercuric acetate (PMA) or the combination of PMA and calcium ionophore. The promoter region between positions -97 and +43 showed responsiveness to low-dose X rays. Electrophoretic mobility shift assays demonstrated that the region from 117 to -97 was responsive to irradiation. Transcription factors specifically bound to this sequence showed a dose-dependent response to single doses of X rays between 1 and 8 Gy. Competition analysis indicated that the protein-DNA complexes at this region were related to the nuclear factor of activated T cells (NF-AT). Further confirmation was obtained by the addition of specific antibodies into protein-DNA reactions. For the first time, we have demonstrated that specific DNA binding of NF-ATp at the promoter region from -117 to -97 is involved in transcriptional regulation of the human IL5 gene in response to ionizing radiation. PMID- 9399699 TI - Enhanced neoplastic transformation in an inhomogeneous radiation field: an effect of the presence of heavily damaged cells. AB - In the inhomogeneous radiation field surrounding small beta-particle sources, nonlethally and heavily damaged cells are in proximity, permitting interaction via extracellular signals. This situation is typical of hot particles such as those released during the accident at Chernobyl. Beta-particle-emitting yttrium 90 wires (average energy 934 keV) were employed to investigate radiation-induced neoplastic transformation under these conditions. Integrated 24-h doses ranging from 0 to 750 Gy across the exposure field were applied. At equal levels of toxicity a 10-fold enhancement of neoplastic transformation frequency in C3H 10T1/2 cells was observed in the presence of heavily damaged cells. Homogeneous fields of low-dose-rate beta-particle radiation produced neoplastic transformation frequencies typical for comparable photon exposures reported in the literature. PMID- 9399700 TI - Comparisons of the frequencies and molecular spectra of HPRT mutants when human cancer cells were X-irradiated during G1 or S phase. AB - In an attempt to elucidate mechanisms underlying the variation in radiosensitivity during the cell cycle, mutations in the HPRT gene were selected with 6-thioguanine, quantified and characterized in synchronous human bladder carcinoma cells (EJ30-15) that were irradiated in G1 or S phase with 3 or 6 Gy. Synchronous cells were obtained by mitotic selection, with approximately 98% of the cells in G1 phase when they were irradiated after 3 h of incubation, and 75% in S phase when they were irradiated after 14 h of incubation. The mutant frequencies were approximately 4-fold higher (P < 0.01) when cells were irradiated in G1 phase compared with S phase, and the lowest frequency (1.5 x 10( 5) for 3 Gy during S phase) was approximately 10-fold higher than the spontaneous frequency. Exon analysis by multiplex polymerase chain reaction was performed on DNA isolated from each independent mutant. The different types of mutants were categorized as class 1, which consisted of base-pair changes or small deletions less than 20 bp; class 2, which consisted of deletions greater than 20 bp but with one or more HPRT exons present; and class 3, which consisted of deletions encompassing the entire HPRT gene and usually genomic markers located 350-750 kbp from the 5' end of the gene and/or 300-1400 kbp from the 3' end. A "hotspot" for class 2 deletions was observed between exons 6 and 9 (P < 0.01). For cells irradiated during G1 phase, the percentages for the different classes (total of 78 mutants) were similar for 3 and 6 Gy, with a selective induction of class 3 mutants (34-38%) compared with spontaneous mutants (3%, total 20). When S-phase cells were irradiated with 3 Gy, there were fewer class 1 mutants (21%, total 37) than when cells were irradiated in G1 phase with 3 Gy (45%, total 42) (P < 0.01). The greatest change was observed when the dose was increased in S phase from 3 Gy to 6 Gy (total of 43 mutants), with the frequency of class 2 mutants decreasing dramatically from 30% to 1% (P < 0.005). A similar decrease in class 2 mutants with an increase in dose has been observed by others in asynchronous cultures of normal human fibroblasts. We hypothesize that these differences occur because: (a) there is more error-free repair of double-strand breaks (DSBs) during S than G1 phase; (b) a single DSB within the HPRT gene causes a class 2 mutation or a certain percentage of class 1 mutations, while two DSBs, with one in each approximately 1-Mbp region 5' and 3' of the gene, cause a class 3 mutation; and (c) a repair process that is induced when the dose during S phase is increased from 3 to 6 Gy results in a preferential decrease in class 2 mutations. PMID- 9399701 TI - Higher-order chromatin structure-dependent repair of DNA double-strand breaks: modeling the elution of DNA from nucleoids. AB - A possible relationship between the repair of DNA double-strand breaks (DSBs) and their distribution within higher-order chromatin (Johnston and Bryant, Int. J. Radiat. Biol. 66, 531-536, 1994) has recently been demonstrated. Radiosensitive cells deficient for components of the DNA-dependent protein kinase DSB repair pathway exhibited a particular failure in the rejoining of DSBs occurring as multiples within looped DNA structures. Here, a Poisson-based model of induction of DSBs and elution of DNA from residual nuclear structures is presented. By applying this model to cells of a panel of human and rodent cell lines, a mean of 1.6 Mbp for the size of the relevant looped structures was obtained. Such large chromatin structures are of the same magnitude as those observed by functional mapping of interphase and mitotic chromosome structure, nucleoid sedimentation and the "replicon clusters" apparent during DNA replication. This work supports the hypotheses that (1) such structures are critical targets for induction of DSBs and (2) the distribution of damage within these domains may be a factor in the response and sensitivity of mammalian cells to ionizing radiation. PMID- 9399702 TI - Temperature dependence of radiation-induced DNA-protein crosslinks formed under hypoxic conditions. AB - Recently, we demonstrated that the oxygen dependence of the formation of DNA protein crosslinks (DPCs) in irradiated mammalian cells measured by the alkaline elution technique is the mirror image of the oxygen dependence of radiation induced cell killing. Consequently, these radiation-induced DPCs could be used to detect hypoxic cells or estimate the hypoxic fraction of cells in solid tumors. Although several techniques, including alkaline elution, gas chromatography/mass spectrometry (GC/MS) and nitrocellulose filter binding, have been used to measure radiation-induced DPCs, the published reports suggest that the characteristics of these DPCs may depend on both the type of sample irradiated (cellular compared to model systems, oxygenated compared to hypoxic, etc.) and the technique used to measure these radiation-induced DPCs. In the present study, the radiation-induced DPCs measured by our alkaline elution technique with and without proteinase K in the lysis solution were characterized by studying the dependence of their formation on temperature in hypoxic rat 9L brain tumor cells. Exponentially growing 9L rat brain tumor cells were rendered hypoxic at 4 degrees C or at 37 degrees C and then irradiated with either 7.5 Gy or 15 Gy. The cells were trypsinized at 4 degrees C, either immediately after the irradiation or after one half-time of strand break repair at 37 degrees C. The results demonstrated that the radiation-induced DPCs produced in 9L cells under hypoxic conditions, measured by our alkaline elution technique after low to moderate radiation doses, required metabolism for their formation, unlike the radiation-induced DPCs reported by others using the GC/MS or nitrocellulose filter binding technique. PMID- 9399703 TI - The involvement of topoisomerase I in the induction of DNA-protein crosslinks and DNA single-strand breaks in cells of ultraviolet-irradiated human and frog cell lines. AB - Exposure of GM 4390 human skin fibroblasts and ICR 2A frog cells to 10 kJ m(-2) of ultraviolet B (UVB) radiation resulted in the formation of DNA-protein crosslinks (DPCs) and DNA single-strand breaks (SSBs). However, upon incubation, there were rapid increases in the yields of both DPCs and SSBs. An enhancement in these DNA alterations was detected within 12 min after irradiation and their levels continued to rise by 5-8-fold within 15 h after exposure to UV radiation. Using an antibody-based assay that measures covalent complex formation between topoisomerase (topo) I and genomic DNA, it was found that topo I is one of the proteins involved in these DPCs induced by UV radiation. The levels and rate of increase of topo I-DNA covalent complexes were similar to the UV-radiation dependent formation of DPCs and SSBs. A UV-radiation-sensitive mutant frog cell line, DRP 153, was also examined and was found to be deficient in this induction of DPCs and SSBs by UV radiation. When these cells were transfected with the human SUVCC3 gene, the resulting transformant displayed kinetics for the induction of DPCs and SSBs similar to the human and parental frog cells. However, human topo I was not defected in the transformed frog cells, indicating that SUVCC3 does not encode topo I. It is likely that SUVCC3 encodes an associated enzymatic activity which permits normal stimulation of topo I-DNA covalent complexes in UV-irradiated cells. PMID- 9399704 TI - Coumarin-3-carboxylic acid as a detector for hydroxyl radicals generated chemically and by gamma radiation. AB - Coumarin-3-carboxylic acid (3-CCA) was used as a detector for hydroxyl radicals (.OH) in aqueous solution. The .OH was generated by gamma irradiation or chemically by the Cu2+-mediated oxidation of ascorbic acid (ASC). The excitation and emission spectra of 3-CCA, hydroxylated either chemically or by gamma irradiation, were nearly identical to those of an authentic 7-hydroxycoumarin-3 carboxylic acid (7-OHCCA). The pH-titration curves for the fluorescence at 450 nm (excitation at 395 nm) of 3-CCA, hydroxylated either chemically or by gamma radiation, were also identical to those of authentic 7-OHCCA (pK = 7.4). Time resolved measurements of the fluorescence decays of radiation- or chemically hydroxylated 3-CCA, as well as those of 7-OHCCA, indicate a monoexponential fit. The fluorescence lifetime for the product of 3-CCA hydroxylation was identical to that of 7-OHCCA (approximately 4 ns). These data, together with analysis of end products by high-performance liquid chromatography, show that the major fluorescent product formed by radiation-induced or chemical hydroxylation of 3 CCA is 7-OHCCA. Fluorescence detection of 3-CCA hydroxylation allows real-time measurement of the kinetics of .OH generation. The kinetics of 3-CCA hydroxylation by gamma radiation is linear, although the kinetics of 3-CCA hydroxylation by the Cu2+-ASC reaction shows a sigmoid shape. The initial (slow) step of 3-CCA hydroxylation is sensitive to Cu2+, but the steeper (fast) step is sensitive to ASC. Analysis of the kinetics of 3-CCA hydroxylation shows a diffusion-controlled reaction with a rate constant 5.0 +/- 1.0 x 10(9) M(-1) s( 1). The scavenging of .OH by 3-CCA was approximately 14% for chemical generation with Cu2+-ASC and approximately 50% for gamma-radiation-produced .OH. The yield of 7-OHCCA under the same radiation conditions was approximately 4.4% and increased linearly with radiation dose. The 3-CCA method of detection of .OH is quantitative, sensitive, specific and therefore accurate. It has an excellent potential for use in biological systems. PMID- 9399705 TI - A semi-empirical approach to the evaluation of the relative biological effectiveness of therapeutic proton beams: the methodological framework. AB - We address the problem of the evaluation of the relative biological effectiveness (RBE) of therapeutic proton beams for cell inactivation. We consider a general approach to the evaluation of the lethal effect of protons which is applicable to different situations, including those in which an extended Bragg peak is obtained via a modulated beam. Our approach combines two kinds of information: (1) the experimental results available in the literature for the response of Chinese hamster V79 cells to monoenergetic beams and (2) the energy spectrum of the beam in the target volume, computed through a Monte Carlo algorithm. We have applied this method to a simple Bragg peak produced by a broad-field proton beam of about 70 MeV initial energy, such as those that, after attenuation, are typically used for treatment of ocular tumors. We have found that the RBE increases with depth, even beyond the Bragg peak, up to a value close to 2, when evaluated at the same surviving fraction as that resulting after exposure to 2 Gy of X rays. PMID- 9399706 TI - Microdosimetry of astatine-211 using histological images: application to bone marrow. AB - A method is presented for calculating the small-scale dosimetry of 211At in red bone marrow using chord-length distributions obtained from digitized histological images. This study used histological samples of bone marrow from beagle dogs to convey morphological information about cell conglomerations within bone marrow. Two 211At activity distributions were considered within the extracellular fluid and the surface of red bone marrow cells. Results confirmed the influence of cell conglomeration and activity distribution in determining the microdosimetry of red bone marrow. Average S* values of 1.6 x 10(-9) and 1.90 x 10(-9) Gy g Bq(-1) s( 1) were calculated for activity distributions located within the extracellular fluid and the surface of red bone marrow cells, respectively. The cumulated activity required to reduce survival probability to 0.37 also was calculated as a function of cell sensitivity for both activity distributions. The activity distribution on the cell surface resulted in a higher cell-killing efficiency, requiring a lower activity concentration of approximately 25% when compared with activity located in the extracellular fluid. Of relevance to potential clinical studies with 211At, the probability for zero hits for red bone marrow cells was > 10% for cumulated activities of less than 5.5 x 10(8) Bq s g(-1) in bone marrow. PMID- 9399708 TI - Measurement of DNA damage after exposure to electromagnetic radiation in the cellular phone communication frequency band (835.62 and 847.74 MHz). AB - Mouse C3H 10T1/2 fibroblasts and human glioblastoma U87MG cells were exposed to cellular phone communication frequency radiations to investigate whether such exposure produces DNA damage in in vitro cultures. Two types of frequency modulations were studied: frequency-modulated continuous-wave (FMCW), with a carrier frequency of 835.62 MHz, and code-division multiple-access (CDMA) centered on 847.74 MHz. Exponentially growing (U87MG and C3H 10T1/2 cells) and plateau-phase (C3H 10T1/2 cells) cultures were exposed to either FMCW or CDMA radiation for varying periods up to 24 h in specially designed radial transmission lines (RTLs) that provided relatively uniform exposure with a specific absorption rate (SAR) of 0.6 W/kg. Temperatures in the RTLs were monitored continuously and maintained at 37 +/- 0.3 degrees C. Sham exposure of cultures in an RTL (negative control) and 137Cs gamma-irradiated samples (positive control) were included with every experiment. The alkaline comet assay as described by Olive et al. (Exp. Cell Res. 198, 259-269, 1992) was used to measure DNA damage. No significant differences were observed between the test group exposed to FMCW or CDMA radiation and the sham-treated negative controls. Our results indicate that exposure of cultured mammalian cells to cellular phone communication frequencies under these conditions at an SAR of 0.6 W/kg does not cause DNA damage as measured by the alkaline comet assay. PMID- 9399707 TI - Measurement of DNA damage after exposure to 2450 MHz electromagnetic radiation. AB - Recent reports suggest that exposure to 2450 MHz electromagnetic radiation causes DNA single-strand breaks (SSBs) and double-strand breaks (DSBs) in cells of rat brain irradiated in vivo (Lai and Singh, Bioelectromagnetics 16, 207-210, 1995; Int. J. Radiat. Biol. 69, 513-521, 1996). Therefore, we endeavored to determine if exposure of cultured mammalian cells in vitro to 2450 MHz radiation causes DNA damage. The alkaline comet assay (single-cell gel electrophoresis), which is reportedly the most sensitive method to assay DNA damage in individual cells, was used to measure DNA damage after in vitro 2450 MHz irradiation. Exponentially growing U87MG and C3H 10T1/2 cells were exposed to 2450 MHz continuous-wave (CW) radiation in specially designed radial transmission lines (RTLs) that provided relatively uniform microwave exposure. Specific absorption rates (SARs) were calculated to be 0.7 and 1.9 W/kg. Temperatures in the RTLs were measured in real time and were maintained at 37 +/- 0.3 degrees C. Every experiment included sham exposure(s) in an RTL. Cells were irradiated for 2 h, 2 h followed by a 4-h incubation at 37 degrees C in an incubator, 4 h and 24 h. After these treatments samples were subjected to the alkaline comet assay as described by Olive et al. (Exp. Cell Res. 198, 259-267, 1992). Images of comets were digitized and analyzed using a PC-based image analysis system, and the "normalized comet moment" and "comet length" were determined. No significant differences were observed between the test group and the controls after exposure to 2450 MHz CW irradiation. Thus 2450 MHz irradiation does not appear to cause DNA damage in cultured mammalian cells under these exposure conditions as measured by this assay. PMID- 9399709 TI - Merril Eisenbud (1915-1997). PMID- 9399710 TI - Prognostic significance of myocardial ischemia detected by ambulatory electrocardiography, exercise treadmill testing, and electrocardiogram at rest to predict cardiac events by one year (the Asymptomatic Cardiac Ischemia Pilot [ACIP] study) AB - Myocardial ischemia identified by ambulatory electrocardiography (AECG), exercising treadmill testing, (ETT), or 12-lead electrocardiogram at rest is associated with an adverse prognosis, but the effect of improving these ischemic manifestations by treatment on outcome is unknown. The Asymptomatic Cardiac Ischemia Pilot (ACIP) study was a National Heart, Lung, and Blood Institute funded study to determine the feasibility of conducting a large-scale prognosis study and to assess the effect of 3 treatment strategies (angina-guided strategy, AECG ischemia-guided strategy, and revascularization strategy) in reducing the manifestations of ischemia as indicated by AECG and ETT. The study cohort for this database study consisted of 496 randomized patients who performed the AECG, ETT, and 12-lead electrocardiogram at rest at both the qualifying and week 12 visits. The effect of modifying ischemia by treatment on the incidence of cardiac events (death, myocardial infarction, coronary revascularization procedure, or hospitalization for an ischemic event) at 1 year was examined. In the 2 medical treatment groups (n = 328) there was an association between the number of ambulatory electrocardiographic ischemic episodes at the qualifying visit and combined cardiac events at 1 year (p = 0.003). In the AECG ischemia-guided patients there was a trend associating greater reduction in the number of ambulatory electrocardiographic ischemia episodes with a reduced incidence of combined cardiac events (r = -0.15, p = 0.06). In the revascularization strategy patients this association was absent. In the medical treatment patients the exercise duration on the baseline ETT was inversely associated with an adverse prognosis (p = 0.02). The medical treatment strategies only slightly improved the exercise time and the exercise duration remained of prognostic significance. In the revascularization group strategy patients this association was absent. Thus, myocardial ischemia detected by AECG and an abnormal ETT are each independently associated with an adverse cardiac outcome in patients subsequently treated medically. PMID- 9399711 TI - Improved specificity of transesophageal dobutamine stress echocardiography compared to standard tests for evaluation of coronary artery disease in women presenting with chest pain. AB - The detection of coronary artery disease (CAD) by noninvasive methods has been hindered in women by the high rate of false-positive results. To determine the feasibility and accuracy of transesophageal dobutamine stress echocardiography for identification of CAD in women, we studied 84 patients (age 51 +/- 11 years) who underwent symptom-limited exercise treadmill testing, exercise thallium-201 scintigraphy, and coronary angiography for evaluation of anginal chest pain. Of the 84 patients, 62 had normal coronary arteries or nonsignificant coronary lesions, and 22 had significant stenosis of > or = 1 major coronary artery. During treadmill exercise, repolarization changes were observed in 16 of 21 patients with CAD and in 19 of 60 patients with normal coronary arteries. With thallium scintigraphy, a reversible defect was observed in 19 of 22 patients with CAD and in 12 of 60 patients with normal coronary arteries. Regional wall motion abnormalities during dobutamine infusion developed in 18 of 22 patients with CAD and in none of the 62 patients with normal coronary arteries. All 3 tests had similar sensitivity for detection of CAD (76% for exercise treadmill test, 86% for thallium scintigraphy, and 82% for transesophageal dobutamine stress echocardiography). However, transesophageal dobutamine stress echocardiography had significantly higher specificity than the other 2 tests (100% vs 68% for exercise treadmill test and 80% for thallium scintigraphy; p = 0.0001). Thus, transesophageal dobutamine stress echocardiography is accurate for evaluation of CAD among women presenting with chest pain; its use should be considered when more conventional tests are equivocal or technically suboptimal. PMID- 9399712 TI - Variations of remodeling in response to left main atherosclerosis assessed with intravascular ultrasound in vivo. AB - Histopathologic studies have demonstrated that vessels enlarge to compensate for an increase in plaque burden; this has been confirmed in vivo using intravascular ultrasound (IVUS). The initial studies suggested a biphasic course of lesion formation with (1) preservation of lumen dimensions up to a plaque burden of approximately 40%, and (2) luminal narrowing as plaque burden further increases. In this study, we used IVUS and angiography to assess the extent of left main (LM) atherosclerosis in 107 patients undergoing catheter-based procedures of the left anterior descending or left circumflex coronary arteries. Using IVUS, atherosclerotic plaques were found in all LM arteries, but only 26 (24%) had varying degrees of luminal narrowing on the angiogram. Nevertheless, there was an inverse relation (r = -0.62, p <0.0001) between the minimal lumen area and the plaque burden (i.e., plaque + media divided by total vessel area) that was not restricted to plaque burden values >40% (or >30%), but persisted at plaque burden values of 20% to 40%. In addition, LM arteries with a plaque burden <40% had a similar total vessel area as did LM arteries with a plaque burden > or =40% (22.9 +/- 6.1 vs 21.8 +/- 4.8 mm2, p = 0.30). These data suggest that lumen dimensions may not be preserved even if plaque occupies no more than 20% to 40% of the total vessel area. Thus, there is more variation in remodeling response during earlier stages of plaque accumulation within the LM artery than is commonly suggested. PMID- 9399713 TI - Gender differences in the accuracy of dobutamine stress echocardiography for the diagnosis of coronary artery disease. AB - The accuracy of dobutamine stress echocardiography (DSE) for the diagnosis of coronary artery disease (CAD) has not been yet evaluated in women. We studied the effect of gender on the accuracy of DSE for the diagnosis of CAD in 306 consecutive patients (210 men and 96 women) with limited exercise capacity and suspected myocardial ischemia who underwent coronary angiography within 3 months of DSE. There were no serious complications during DSE. Men had a higher prevalence of nonsustained ventricular tachycardia (7% vs 0.03%, p <0.05) and supraventricular tachycardia (9% vs 0.03%, p <0.05) during the test compared with women. Peak stress rate-pressure product was not different in men and women (18,140 +/- 4,187 vs 18,543 +/- 4,223). Significant CAD (> or =50% luminal diameter stenosis) was present in 171 men (81%) and in 62 women (65%, p <0.005). The sensitivity, specificity, and accuracy of ischemic pattern at DSE for the diagnosis of significant CAD were 76% (confidence interval [CI] 67 to 84), 94% (CI 89 to 99), and 82% (CI 75 to 90) in women and 73% (CI 67 to 79), 77% (CI 71 to 83), and 74% (CI 68 to 80) in men, respectively. Overall specificity was higher in women than in men (p <0.05). Regional accuracy of DSE was significantly higher in women than in men in the 3 arterial regions (84% [CI 79 to 88] vs 75% [CI 72 to 79], p <0.005). It is concluded that DSE is a safe and feasible method for the diagnosis of CAD in women. The overall specificity and the regional accuracy of DSE are higher in women than in men. Further studies are required to evaluate the functional significance of these findings and their reproducibility in different patient populations. PMID- 9399714 TI - Recanalization of total coronary occlusions using a laser guidewire (the European TOTAL Surveillance Study). AB - The success rates of coronary angioplasty for the treatment of chronic total occlusions are less favorable than for coronary stenosis. Therefore, a new laser guidewire (LW) was designed to facilitate the crossing of chronic total occlusions. We report on the results of a European multicenter surveillance study, evaluating the laser guidewire performance. Between May 1994 and July 1996, 345 patients (age 59 +/- 10 years, 291 men) with chronic total occlusions were enrolled in 28 European centers. The median age of occlusion was 29 weeks (range 2 to 884), the occlusion length 19 +/- 10 mm. LW recanalization was successful in 205 patients (59%/). LW perforation occurred in 73 patients (21%), with hemodynamic consequences in 4 (1%). There were no deaths, emergency coronary artery bypass graft surgery, or Q-wave myocardial infarctions. In a multivariate regression analysis an occlusion age of <40 weeks (p = 0.001, RR = 1.34) and an occlusion length <30 mm (p = 0.01, RR = 1.59) were independent predictors of success. Results indicate that the LW is an effective and safe tool in the treatment of chronic total occlusion refractory to conventional guidewires. PMID- 9399715 TI - Treatment of in-stent coronary restenosis by excimer laser angioplasty. AB - We evaluated the efficacy and safety of excimer laser angioplasty (ELCA) with adjunctive balloon angioplasty in patients with restenotic or occluded coronary stents. ELCA was performed in 70 patients (60 +/- 9 years), who had previously been treated with Micro Stents (n = 65), Palmaz-Schatz (n = 38), Wiktor, NIR, Freedom, and Multi-Link stents (n = 1 each). Restenosis (> or =50% diameter stenosis) was documented in 90 stents, another 17 stents were occluded. Laser energy was delivered to the lesions with catheters 1.4, 1.7 (eccentric), and 2.0 mm in diameter. Procedural success was controlled by intravascular ultrasound in a subgroup. Laser catheters crossed all restenotic or occluded stents and decreased diameter stenosis from 80 +/- 13% to 44 +/- 11% (p <0.001). Adjunctive balloon angioplasty further reduced diameter stenosis to 13 +/- 13% (p <0.001). In 13 patients with 21 stents, serial intravascular ultrasound imaging revealed a reduction of plaque area within the stent by 34 +/- 22% (from 4.2 +/- 1.8 mm2 to 2.7 +/- 1.1 mm2) after ELCA and a reduction by 65 +/- 16% (to 1.5 +/- 0.7 mm2) after balloon angioplasty (p <0.01). There were 4 patients with an increase of creatine kinase levels, 8 patients with major dissections (in 7 patients they were related to adjunctive balloon angioplasty), 1 patient with distal embolization, 2 with minor perforations, and 1 patient with stent dislocation. Reintervention during hospitalization was necessary in 3 patients. ELCA is an efficient and safe technique to debulk tissue in restenotic lesions and total occlusions within stents. The incidence of procedure related complications was low. PMID- 9399716 TI - Relation of coronary artery disease to atherosclerotic disease in the aorta, carotid, and femoral arteries evaluated by ultrasound. AB - This prospective study was conducted to correlate the presence of angiographically significant coronary artery disease (CAD) and atherosclerotic disease in the aorta, carotid, and femoral arteries as measured by ultrasound. One hundred two consecutive patients admitted for coronary angiography for suspected CAD participated in the study. All patients underwent transesophageal echocardiography for the evaluation of thoracic aortic atherosclerosis and B-mode ultrasound for evaluation of carotid and femoral atherosclerosis. Intimal-medial thickness > 1 mm in the thoracic aorta or peripheral vessels was considered as evidence of atherosclerosis. Patients with CAD (n = 64) had a significantly higher incidence of atherosclerotic plaques in the thoracic aorta, carotid, and femoral arteries than subjects with normal coronary arteries: 91%, 72%, 77% vs 31%, 47% and 42%, respectively. Extracoronary plaque was a stronger predictor of CAD than conventional risk factors. Evidence of plaque in patients younger than median age (64 years) had a higher specificity than in patients above median age (77% vs 40%, respectively, p <0.0001). Plaque score of the extracardiac vessels was significantly higher in patients with multivessel CAD than in patients with 1 vessel CAD disease and in subjects with normal coronary arteries (p <0.001). Thus, atherosclerotic plaques in the aortic and femoral arteries and, to a lesser extent, in the carotid arteries are strong predictors of CAD. PMID- 9399717 TI - Influence of physical activity on 24-hour measurements of heart rate variability in patients with coronary artery disease. AB - This study assessed the influence of physical activity on time domain variables of heart rate variability (HRV) during 24-hour electrocardiographic registrations. Changes in time domain variables of HRV (in particular SDNN) obtained from Holter recordings were proven as strong predictors of cardiac events in patients with coronary artery disease. Although 24-hour measurements of HRV recordings are a standard technique, little is known about the effects of the environment during the registration period. This applies especially to the type and nature of physical activity. In a prospective study, 106 patients with angiographically proven coronary artery disease were randomized into 2 groups. Group 1 consisted of 54 patients with recordings under normal daily physical activities. Group 2 consisted of 52 patients who were immobilized during the recording. Both groups were comparable concerning clinical parameters. The results of 24-hour measurements of HRV with analysis of time domain variables (SDNN, SDANN, SDNN index, rMSSD, and pNN50) were compared among the 2 patients groups, and with a healthy control group. Comparison of immobilized patients with healthy controls showed statistically significant differences of all HRV parameters (p <0.01). However, when comparing the activity group with healthy controls, none of the parameters showed any significant differences. Comparison of the subgroups revealed statistically significant differences of the parameters SDNN, SDANN (p <0.01), and borderline results for rMSSD and pNN50 (p = 0.05). Our results indicate that time domain variables of HRV calculated from 24-hour recordings are significantly influenced by the level of physical activity and the upright posture during registration. This methodologic aspect has to be considered, especially if HRV measurements are used as prognostic markers in patients with coronary heart disease. PMID- 9399718 TI - Radiofrequency catheter ablation for paroxysmal supraventricular tachycardia in children and adolescents without structural heart disease. Pediatric EP Society, Radiofrequency Catheter Ablation Registry. AB - Since 1990, management options available for children with paroxysmal supraventricular tachycardia (PSVT) have included radiofrequency catheter ablation (RCA). To determine the efficacy and safety of the procedure and to maintain a database for long-term follow-up, the Pediatric Electrophysiology Society began a Pediatric RCA Registry on January 1, 1991, to which 46 centers have submitted data from 4,135 total children and adolescents (patient age 0.1 to 20.9 years) who underwent 4,651 RCAs (through September 15, 1996). Of the 88% with a structurally normal heart, PSVT mechanisms (n = 4,030) included 3,110 accessory pathways and 920 atrioventricular node reentry tachycardia (AVNRT) during 3,653 procedures for 3,277 patients. During the 7 years of the Registry, analysis of indications for the procedure has shown a gradual shift. During the first year of the Registry for this PSVT group, "medically refractory tachycardia" was listed as the indication for 44% and "patient choice" was listed as 33%, compared with 29% and 58%, respectively, for the years 1995 to 1996 (p <0.005). Registry results were: 90% immediate success for accessory pathways (95% for left lateral; 87% for septal; 86% for right free wall) and 96% for AVNRT; mean fluoroscopy time 47.6 +/- 40 SD minutes; procedure time 257 +/- 157 SD minutes; major complication rate at the time of the procedure 3.2%. Procedure related deaths included 1 immediate and 3 at 2, 12 and 68 weeks after the procedure (2 were infants). Follow-up revealed 77% and 71% freedom from recurrence at 3 years for accessory pathways AVNRT, respectively, and rare (<1%) detection of additional complications. RCA has evolved into a standard management option for PSVT in children with a structurally normal heart. RCA for children and adolescents should be recommended after consideration of the procedural risk/benefit compared with that of other management options, the natural history, and individual tolerance/symptoms related to PSVT. PMID- 9399719 TI - Comparison of power- and temperature-guided radiofrequency modification of the atrioventricular node. Polaris Investigator Group. AB - The purpose of this study was to compare the performance and clinical outcome of radiofrequency ablation of the substrate of atrioventricular (AV) nodal reentrant tachycardia (AVNRT) when guided by power output or temperature monitoring. Two sequential multicenter studies of power-controlled and open-loop, temperature controlled radiofrequency ablation were analyzed in 171 patients undergoing AV node modification for the treatment of AVNRT. After successful ablation of AVNRT, complete elimination of slow AV node pathway function was accomplished more often with than without temperature monitoring (92% vs 69%, p = 0.005). Greater power was delivered to each patient with than without temperature monitoring (median 47 W, range 10 to 57, vs median 35 W, range 5 to 68, p = 0.001). Acute elimination of tachycardia (100% vs 96%), 3-month recurrence (6% vs 8%), procedural times (162 vs 170 minutes), fluoroscopy times (24.6 vs 29.5 minutes), complications (6% vs 3%), and catheter removals to check for coagulum (8% vs 6%) did not differ between patients treated with and without temperature monitoring, respectively. Power- and temperature-controlled radiofrequency techniques are highly successful with low complication rates for slow pathway ablation. Temperature monitoring may allow the safe delivery of more power, and the more complete elimination of slow AV node pathway function. PMID- 9399720 TI - Effect of reproducibility of baseline arrhythmia induction on drug efficacy predictions and outcome in the Electrophysiologic Study Versus Electrocardiographic Monitoring (ESVEM) trial. AB - Spontaneous variability over time in the ease of induction of ventricular arrhythmias may mimic a drug effect and affect the predictive value of drug therapy guided by programmed stimulation. We assessed the effect of baseline reproducibility of arrhythmia induction on the incidence and accuracy of drug efficacy predictions in the Electrophysiologic Study Versus Electrocardiographic Monitoring (ESVEM) trial. Patients with sustained ventricular tachyarrhythmias induced twice during baseline electrophysiologic testing with the same stimulation technique, i.e., induced at the same pacing site with the same drive cycle length and number of extrastimuli, were identified from the ESVEM database. These patients with highly reproducible arrhythmia induction were compared to those with less reproducible arrhythmias. Of 473 randomized patients with reproducibility data, 313 (66%) had highly reproducible arrhythmias. In patients randomized to electrophysiologic testing, baseline arrhythmia reproducibility did not affect the incidence of drug efficacy predictions (70 of 157 [45%], drug efficacy predictions in patients with highly reproducible arrhythmias vs 34 of 79 [43%] with less reproducible arrhythmias, p = 0.890). Drug efficacy predictions obtained by electrophysiologic testing in patients with highly reproducible arrhythmias were not associated with decreases in arrhythmia recurrence (p = 0.202), all-cause mortality (p = 0.301), cardiac death (p = 0.358), or arrhythmic death (p = 0.307) compared to those with less reproducible arrhythmias. Analysis of patients with highly reproducible sustained monomorphic ventricular tachycardia led to similar results. In the ESVEM trial, most patients had highly reproducible arrhythmia induction during baseline electrophysiologic testing. Reproducibility of arrhythmia induction in the baseline state had no effect on the incidence or accuracy of drug efficacy predictions. PMID- 9399721 TI - Effect of calcium antagonists on plasma norepinephrine levels, heart rate, and blood pressure. AB - To evaluate the effects of calcium antagonists on sympathetic activity in hypertensive patients, a MEDLINE search for English language articles published between 1975 and May 1996 using the terms calcium antagonists, sympathetic nervous system, and catecholamines was conducted. Clinical studies only reporting the effects of calcium antagonists on blood pressure, heart rate, and plasma norepinephrine (NE) levels in patients with hypertension were included. Data were combined and analyzed according to class of calcium antagonist (dihydropyridine vs nondihydropyridine), their duration of action (short-acting [SA] vs long acting [LA]), and treatment duration. We identified 63 studies involving 1,252 patients. Acutely after single dosing, SA calcium antagonists decreased mean arterial pressure by 13.7 +/- 1.1% and increased heart rate by 13.7 +/- 1.4% and NE levels by 28.6 +/- 2.5%. Change in NE levels correlated with change in heart rate (r = 0.59, p <0.01) and inversely with change in arterial pressure (r = 0.46, p <0.05) in patients taking dihydropyridine calcium antagonists acutely. With sustained therapy, both classes of SA calcium antagonists increased NE levels. Whereas NE levels remained slightly elevated and heart rate unchanged with LA dihydropyridine calcium antagonists, both heart rate and NE levels decreased with LA nondihydropyridine calcium antagonists. SA calcium antagonists stimulate sympathetic activity when given acutely and over the long term, irrespective of their molecular structure. Sympathetic activation is less pronounced with LA dihydropyridine calcium antagonists and decreases with LA nondihydropyridine calcium antagonists. These data offer a possible pathophysiologic explanation for the increase in morbidity and mortality observed in some studies using SA calcium antagonists. PMID- 9399722 TI - Usefulness of low doses of atropine to quantify the vagal stimulus-response relation in patients with congestive heart failure. AB - The response of low doses of atropine is reported to be attenuated in patients with congestive heart failure (CHF). Judging from the main site of action of low doses of atropine, we may be able to assess the functional state of the vagal center in the central nervous system. This study examines the clinical significance of heart rate (HR) response to a low dose of atropine in patients with CHF. Low and high doses of atropine were administered intravenously in 72 patients with CHF. HR after a low (parasympathomimetic) dose injection was assessed by the ratio Rm (minimal HR/basal HR), and after a high (parasympatholytic) dose by the ratio R1 (augmented HR/basal HR). Rm and R1 were related to indexes of CHF. Rm increased with progression of CHF (0.92 +/- 0.03 in New York Heart Association functional class I, 0.98 +/- 0.05 in class II, and 1.00 +/- 0.04 in class III). It also correlated with ejection fraction (r = 0.48, p <0.01) and more importantly, with peak oxygen uptake (r = -0.59, p <0.01). R1 exhibited weak correlation with basal HR (r = -0.33, p <0.05) and ejection fraction (r = 0.31, p <0.05), but had no correlation with other indexes. The vagal center may be already blunted in New York Heart Association class II with respect to increased Rm, which may be related to depressed exercise capacity. A low dose of atropine injection offers a simple and safe method for providing important information on the functional state of the vagal center in the central nervous system in patients with CHF. PMID- 9399723 TI - Time-related trends in the preoperative evaluation of patients with valvular stenosis. AB - To investigate time-related trends in the use of preoperative invasive hemodynamics in patients with pure valvular stenosis, the preoperative evaluations and preoperative echocardiograms of consecutive patients who underwent aortic or mitral valve surgery from 1986 to 1994 at the Cleveland Clinic Foundation were reviewed. The study group consisted of 1,985 patients, 1,476 with aortic stenosis and 509 with mitral stenosis. Preoperative cardiac catheterization was performed in 1,456 patients with aortic stenosis (99%) and 488 with mitral stenosis (96%). Measurement of invasive hemodynamics (including transvalvular gradients and estimated valve areas) during catheterization decreased over time both in patients with aortic (from 64% in 1986 to 30% in 1994, test for trend p <0.0001) and mitral stenosis (from 63% in 1986 to 18% in 1994, test for trend p <0.0001). After adjusting for age, gender, and other characteristics, the only predictors of performance of invasive hemodynamics in patients with aortic stenosis were more recent surgery (inverse relation, p = 0.0001) and New York Heart Association class (p = 0.01); in patients with mitral stenosis the only predictor was also more recent surgery (inverse relation, p = 0.0001). Thus, use of preoperative invasive hemodynamics in patients with valvular stenosis has markedly decreased over the last decade. This is an example of how a noninvasive modality can supercede an invasive one, even when surrounding a procedure as fundamentally invasive as valvular heart surgery. PMID- 9399725 TI - Treatment and outcome in silent myocardial ischemia: more pieces of the puzzle. PMID- 9399724 TI - Cardiovascular function before, during, and after the first and subsequent pregnancies. AB - This study was designed to test the hypothesis that the vascular remodeling of pregnancy begins early, persists for at least 1 year after delivery, and is accentuated by a second pregnancy. Serial estimates of heart rate, arterial pressure, left ventricular volumes, cardiac output, and calculated peripheral resistance were obtained before pregnancy, every 8 weeks during pregnancy, and 12, 24, and 52 weeks postpartum in 15 nulliparous and 15 parous women using electrocardiography, automated manometry, and M-mode ultrasound. During pregnancy, body weight increased 14.5 +/- 1.8 kg and returned to prepregnancy values 1 year postpartum. Heart rate peaked at term 15 +/- 1 beat/min above prepregnancy levels (57 +/- 1 beat/min). Mean arterial pressure reached its nadir (-6 +/- 1 mm Hg) at 16 weeks, returning to baseline at term. The increases in left ventricular volumes and cardiac output (2.2 +/- 0.2 L/min) peaked at 24 weeks as did the 500 +/- 29 dynes x cm x s(-5) decrease in peripheral resistance, and their magnitude was significantly greater in the parous women. Postpartum they gradually returned toward baseline but remained significantly different from prepregnancy values in both groups at 1 year. We conclude that cardiovascular adaptations to the initial pregnancy begin early, persist postpartum, and appear to be enhanced by a subsequent pregnancy. We speculate that persistence of these changes may lower cardiovascular risk in later life. PMID- 9399726 TI - Coronary angioplasty induces a systemic inflammatory response. AB - C-reactive protein (CRP) levels increased more than sixfold above baseline when measured 48 hours after elective percutaneous transluminal coronary angioplasty (PTCA) in patients without underlying inflammatory conditions and did not change significantly in controls undergoing coronary angiography. Only 3 of the 42 PTCA patients had clinical restenosis and underwent target vessel revascularization during the 6-month follow-up, but 2 of the 3 had very high CRP levels 48 hours after the procedure. PMID- 9399727 TI - Relation between arteriosclerosis in the coronary and renal arteries. AB - This study analyzes the severity of coronary artery disease in terms of the severity of renal artery disease in 609 patients undergoing coronary and renal angiography. The presence of renal artery disease of any severity is strongly suggestive of advanced coronary artery disease. PMID- 9399728 TI - Predictive accuracy of echocardiographic response of mildly dyssynergic myocardial segments to low-dose dobutamine. AB - Low-dose dobutamine echocardiography has a high sensitivity for predicting functional recovery after revascularization. In segments with mild wall motion abnormalities, the specificity of the technique is rather low, suggesting overestimation of functional recovery in these segments. PMID- 9399729 TI - Insulin resistance in patients with familial combined hyperlipidemia and coronary artery disease. AB - The minimum model modified by the administration of insulin provides an objective and relatively easily measured index of peripheral sensitivity to insulin which was significantly lower (p <0.02) in familial combined hyperlipidemia (FCH) with ischemic heart disease (IHD) than in FCH without IHD and in control subjects (1.2 +/- 0.6, 1.9 +/- 1.0, 2.9 +/- 1.2 x 10(-4) mU/L/ min, respectively). In patients with FCH, insulin resistance explains, at least in part, their metabolic alterations (hypertension, abnormal glucose tolerance, hyperinsulinemia) and elevated IHD. PMID- 9399730 TI - Antitachycardia pacing in patients with implantable cardioverter-defibrillators: inverse circadian variation of therapy success and acceleration. AB - We analyzed spontaneous ventricular tachycardias treated by antitachycardia pacing during long-term follow-up in 138 recipients of an implantable cardioverter-defibrillator. An inverse circadian variation of the antitachycardia pacing termination and acceleration rates with the worst antitachycardia pacing success during the time period with the highest episode frequency (morning hours) was demonstrated. PMID- 9399731 TI - Tolerability of internal low-energy shock strengths currently needed for endocardial atrial cardioversion. AB - There seems to be no relation between shock strength and patient's tolerability using energy levels currently needed for low-energy internal atrial cardioversion. Every patient felt that the second delivered shock, independent of the amount of energy, was more uncomfortable than the first one, which indicates that psychological conditioning may also play an important role in determining discomfort. PMID- 9399732 TI - Symptomatic conduction system disease in cardiac amyloidosis. AB - Symptomatic conduction system disease in cardiac amyloidosis and its management has been reported infrequently. We report our experience of patients with amyloidosis having symptomatic conduction system disease requiring permanent pacemaker implantation. PMID- 9399733 TI - Immediate reproducibility of tilt-table test results in elderly patients referred for evaluation of syncope or presyncope. AB - We studied the immediate reproducibility of the results of the head-up tilt-table test in elderly patients (age > or =60 years). Twenty-seven consecutive men underwent 51 tests. The overall reproducibility was 98%, including 92% of a positive test and 100% of a negative test. The finding of this study validates the use of intravenous pharmacologic intervention in the elderly population. PMID- 9399734 TI - Value of the electrocardiogram in determining cardiac events and mortality in myotonic dystrophy. AB - Electrocardiograms were recorded at baseline and regular intervals in 53 patients with myotonic dystrophy who were followed for a mean of 6.3 +/- 4.0 years. Patients with cardiac events had a significantly prolonged PR interval (p <0.001), a later age of onset of neuromuscular symptoms (p <0.05), and were older (p <0.005). PMID- 9399735 TI - Effectiveness of five-loop coils to occlude patent ductus arteriosus. AB - Coil occlusion of patent ductus arteriosus with 5-loop coils was undertaken in 10 patients without coil embolizations, and with 90% immediate occlusion and 100% occlusion at follow-up. We conclude that 5-loop coil occlusion of patent ductus arteriosus is safe and effective. PMID- 9399736 TI - Effectiveness of an umbilical artery "snare assisted" approach for critical pulmonary valve stenosis or atresia in the neonate. AB - Thirteen neonates with critical pulmonary valve stenosis/atresia underwent successful transcatheter balloon valvuloplasty using an umbilical artery "snare assisted" approach. This technique simplifies the procedure and avoids femoral artery injury by using the umbilical artery, reduces fluoroscopy exposure, and eliminates the need for a gradational approach which reduces costs. PMID- 9399737 TI - Effects of hormone therapy on inflammatory cell adhesion molecules in postmenopausal healthy women. AB - To investigate the effect of estrogen with antioxidant potential on soluble markers of chronic vascular inflammation, we administered either transdermal 17beta-estradiol 0.1 mg/day (9 women) or 17beta-estradiol 0.1 mg and medroxyprogesterone acetate 2.5 mg/day (11 women) for 1-month treatment in a randomized design, with measurement of cell adhesion molecules. Hormone therapy significantly lowered intercellular adhesion molecules-1 levels by 8% (p = 0.009) and tended to lower E-selectin levels (by 6%, p = 0.096), and VCAM-1 levels (by 4%, p = 0.084). PMID- 9399738 TI - Coronary vasospasm induced during isoproterenol head-up tilt test. PMID- 9399739 TI - Use of telemetered permanent pacemaker intracardiac electrograms to diagnose ventricular tachycardia. PMID- 9399740 TI - Transcoronary alcohol ablation of infundibular hypertrophy in patients with idiopathic infundibular pulmonic stenosis. PMID- 9399741 TI - Unresolved issues in gastroesophageal reflux-related ear, nose, and throat problems. PMID- 9399742 TI - Molecular screening: why haven't we started yet? PMID- 9399743 TI - The role of the oral cavity in Helicobacter pylori infection. PMID- 9399744 TI - New trends in liver transplantation for viral hepatitis. PMID- 9399745 TI - Empiric trial of high-dose omeprazole in patients with posterior laryngitis: a prospective study. AB - The optimal management of patients suspected with gastroesophageal reflux-related posterior laryngitis is unclear. History, physical examination, and ambulatory pH monitoring all have significant limitations in identifying patients who will respond to antireflux therapy. OBJECTIVE: To evaluate the merit of empiric omeprazole therapy in patients with posterior laryngitis. METHODS: Twenty-two patients (11 men/11 women, median age 58 yr) with newly diagnosed posterior laryngitis were enrolled. All had persistent laryngeal symptoms for at least 1 month. An empiric trial of omeprazole at 40 mg q.h.s. was given for 8 wk. Four laryngeal symptoms (hoarseness, throat burning/pain, throat clearing, and cough) and four esophageal symptoms (heartburn, regurgitation, dysphagia, and odynophagia) were scored from 0 to 3. Symptom scores were obtained before, 4 wk after, and 8 wk after the start of omeprazole. Patients were classified as responders if they were symptom free or satisfied with results. Omeprazole was stopped in the responders to look for relapse. Ambulatory pH monitoring was performed in patients who did not respond. RESULTS: One patient discontinued omeprazole and withdrew from the study. In the remaining 21 patients, the total laryngeal and esophageal symptom scores significantly improved after empiric omeprazole. Fourteen patients (67%) were classified as responders. Eight patients (38%) had a relapse when omeprazole was stopped. Six patients (29%), interestingly, did not relapse and did not require long-term antireflux therapy. Seven patients (33%) were classified as nonresponders. Ambulatory pH monitoring was abnormal in four of the five patients who agreed to have this test. Increasing the dose of omeprazole to 40 mg b.i.d. provided no additional benefit in the nonresponders. CONCLUSIONS: Empiric omeprazole therapy is a reasonable, initial approach to patients with suspected gastroesophageal reflux-related posterior laryngitis. A significant number of patients do well with a short course of antireflux therapy. Additionally, a third of the patients may not completely respond to intensive medical therapy despite the fact that reflux is documented. PMID- 9399746 TI - Detection of Ki-ras mutations by PCR and differential hybridization and of p53 mutations by SSCP analysis in endoscopically obtained lavage solution from patients with long-standing ulcerative colitis. AB - OBJECTIVES: The goal of this study was the early detection of malignant transformation in patients with long-standing ulcerative colitis; therefore, mutations of the Ki-ras and p53 gene were analyzed in lavage solution and biopsies obtained at surveillance colonoscopy. METHODS: DNA was isolated from 14 patients (nine female, five male) with a history of pancolitis for more than 10 yr. Exon 1 of the Ki-ras gene and exons 5-8 of the p53 gene were amplified via polymerase chain reaction. Mutations of the p53 gene were detected via single strand conformation polymorphism analysis; point mutations of the Ki-ras gene were hybridized on dot blots with oligonucleotides marked with digoxigenin. RESULTS: Wild-type Ki-ras and wild-type p53 were detected in all cases of ulcerative colitis and in four of seven control patients. In two ulcerative colitis patients, a mutation was found in the Ki-ras gene (Gly --> Asp 12 and Gly --> Val 12), and in one patient, a mutation in exon 5 of the p53 gene. Mutations were found only in the lavage fluid, whereas random biopsies were negative. CONCLUSIONS: From colonic lavage fluid, it is possible to extract DNA of sufficient quantity and quality for polymerase chain reaction-based amplification and subsequent analysis via single-strand conformation polymorphism or hybridization. Mutations were found in three of 14 patients with long-standing ulcerative colitis but were not found in controls. The method may be useful for the screening of such patients. PMID- 9399747 TI - Patient preferences and quality of life associated with colorectal cancer screening. AB - OBJECTIVES: The goal of this study was to describe the attitudes of patients toward colorectal cancer screening, colon cancer, and colostomy. METHODS: Using the time trade-off technique, we interviewed four groups of patients at a veterans' hospital: 1) 46 patients with colorectal cancer, 2) 24 patients undergoing screening sigmoidoscopy, 3) 114 subjects participating in a screening colonoscopy study, and 4) 62 patients who have never undergone endoscopic screening for colorectal cancer. Using this technique, we measured quality of life for six scenarios pertaining to screening for colorectal cancer, the patient's current health, colorectal cancer, and colostomy. RESULTS: Unscreened patients were willing to give up significantly more time to avoid screening sigmoidoscopy and colonoscopy (median 91 days and 183 days, respectively) than were patients undergoing screening sigmoidoscopy (median 0 days and 7 days, respectively), screening colonoscopy (median 0 days and 0 days, respectively), or patients with colorectal cancer (median 0 days and 0 days, respectively). Cancer patients rated their current health state lower than volunteers for screening. Colon cancer and colostomy were rated similarly by all four groups. Substantial variation in patient attitudes was present in all groups. CONCLUSIONS: Patients are generally very accepting of endoscopic screening for colorectal cancer. However, decisions regarding recommendations for colorectal cancer screening must take into account the variability in patient preferences. Effective alternative strategies should be available for those whose preferences do not comply with standard recommendations. The effect of patient education and physician recommendations on subjects' attitudes toward screening warrants further investigation. PMID- 9399748 TI - Proton pump inhibitors or histamine-2 receptor antagonists for the prevention of recurrences of erosive reflux esophagitis: a cost-effectiveness analysis. AB - OBJECTIVES: Erosive esophagitis is a recurring condition for which many patients require preventive therapy. If maintenance therapy must be provided, the most cost-effective treatment strategy should be established. We evaluated the costs and benefits associated with three treatment strategies: 1) maintenance therapy with a proton pump inhibitor (PPI) strategy, 2) maintenance therapy with a high dose histamine-2 receptor antagonist (H2RA) strategy, and 3) maintenance therapy with a standard-dose H2RA. If patients experience a symptomatic recurrence on the H2RA strategies, they then receive PPI maintenance. METHODS: We used a cost effectiveness model with a 1-yr time frame; data were obtained from randomized trials of lansoprazole and ranitidine, from case series, and expert opinion. RESULTS: In most situations, the high-dose H2RA strategy is the most costly, yet it is less effective than the PPI strategy. Among the remaining two options, the PPI strategy is more costly and more effective than the standard-dose H2RA strategy, requiring an additional $52-688 per recurrence prevented, depending on drug acquisition costs. The greater the degree to which esophagitis decreases quality of life, the more cost effective is the PPI strategy. For example, with a $50,000 per quality-adjusted life year cost-effectiveness threshold and a market weighted average of drug costs, the PPI strategy appears cost effective for those patients who report that symptoms of esophagitis cause greater than a 9% decrement in quality of life. CONCLUSIONS: The high-dose H2RA strategy is not preferred in terms of either costs or benefits. The PPI strategy appears cost effective relative to the standard-dose H2RA strategy in the following situations: when patients are significantly bothered by esophagitis and in institutional settings where the difference in drug costs between PPIs and H2RAs is small. PMID- 9399749 TI - Coated expandable esophageal stents in the treatment of digestive-respiratory fistulas. AB - BACKGROUND: Malignant digestive-respiratory fistula (DRF) is associated with significant morbidity and mortality. In addition to the other recognized advantages of expandable stents, coated expandable stents can seal off DRF. METHODS: Eight men and five women, mean age 52 yr, with endoscopically and radiographically proven DRF were treated with the coated Wallstent (Schneider). Eleven had dysphagia, 11 postprandial cough, and two required mechanical ventilation. The DRF was proximal in four, mid-esophageal in seven, and distal in two. Two had a normal esophagus and 11 had stricture. RESULTS: Stent placement and DRF obliteration were successful in all. During a median follow-up of 157 days (range 30-423), no recurrent DRF were noted. The median dysphagia score improved from 3.4 to 1.3. Respiratory symptoms were corrected in all. A gastrostomy tube was required in three. The only complications were transient chest pain and foreign body sensation in three patients and constant sensation of belching in one. There was no procedure-related mortality. CONCLUSION: In this small group of patients, the coated Wallstent demonstrated excellent palliation of DRF with minimal morbidity and no mortality. PMID- 9399750 TI - Does chemoradiation therapy increase the incidence of complications with self expanding coated stents in the management of malignant esophageal strictures? AB - BACKGROUND: Expandable stents offer excellent palliation of malignant dysphagia and digestive-respiratory fistula. There are insufficient data regarding factors that may affect the complication rate of expandable stents, but an association between previous treatment with chemotherapy and/or radiation therapy and stent related life-threatening complications has been suggested. METHODS: We retrospectively analyzed our data on 60 patients; in all of them, a coated Wallstent had been successfully placed for malignant dysphagia and/or digestive respiratory fistula. Our objective in this study was to determine the overall complication rate as well as whether previous or ongoing chemoradiation therapy increased the rate of life-threatening complications. RESULTS: Among 21 patients with no previous chemotherapy or radiation therapy, two (9.5%) had life threatening complications (both had bleeding tumors; blood transfusions were required in two and endoscopic hemostasis in one). Among 39 patients who had had either radiation therapy, chemotherapy, or both, life-threatening complications occurred in three (8%). Two of the three had gastrointestinal bleeding (two received blood transfusions, and one had external radiation therapy), and in the third, an esophageal tear was treated with the stent. There was no procedure- or stent-related mortality in either group. CONCLUSIONS: Palliation of malignant dysphagia or digestive-respiratory fistulas with the coated Wallstent in patients with previous chemotherapy and/or radiation therapy is not associated with an increased risk of life-threatening complications. PMID- 9399751 TI - Genetic distinctions between types 1 and 2 autoimmune hepatitis. AB - OBJECTIVES: Our aim was to determine whether alleles affecting susceptibility to type 1 autoimmune hepatitis in the United States occur as commonly in German patients with type 2 disease. METHODS: DNA specimens from 12 German patients with type 2 autoimmune hepatitis were tested for class II alleles of the major histocompatibility complex by polymerase chain reaction using sequence specific primers. Eighty-six American patients with type 1 disease and 102 Caucasoid normal subjects from the United States were tested in a similar manner. RESULTS: American patients with type 1 autoimmune hepatitis had DRB1*03 alleles more commonly than the German patients with type 2 disease (51% vs 17%, p = 0.03) and DRB1*0301 occurred more frequently in the type 1 patients (51% vs 17%, p = 0.03). The frequency of DRB1*04 alleles was also higher in the type 1 patients after exclusion of the DR1*03 alleles (64% vs 20%, p = 0.01). In contrast, patients with type 2 disease more commonly had DRB1*07 (p = 0.003), DRB1*15 (p = 0.004), and DQB1*06 (p = 0.0004). DRB1*07 (p = 0.005), DRB4*01 (p = 0.03), and DQB1*06 (p = 0.03) also occurred more frequently in the type 2 patients from Germany than in the normal subjects from the United States, although none of these frequencies were statistically significant by an adjusted p value. CONCLUSIONS: German patients with type 2 autoimmune hepatitis do not have the same susceptibility alleles as American patients with type 1 disease. Regional differences in prevalence may reflect the genetic profiles of the populations at risk. PMID- 9399752 TI - Safety of topical 5-aminosalicylic acid in pregnancy. AB - OBJECTIVE: To assess the safety and efficacy of topical 5-aminosalicylic acid preparations for therapy of distal colitis during pregnancy. METHODS: Nineteen pregnancies in sixteen consecutive patients with proven distal colitis were identified prospectively at a tertiary care center. All patients were given trials of weaning off medication and all failed. All subjects were on maintenance topical 5-aminosalicylic acid therapy at the time of conception. They were followed throughout their pregnancy and thereafter. Their children were also closely examined and followed by a pediatrician. RESULTS: The mean age at delivery was 25.8 yr, and the mean duration of illness was 4.6 yr. After taking topical therapy, there were no relapses during the pregnancy. There were 19 successful full-term pregnancies with no fetal abnormalities. The mothers and children were followed for more than 6 months. CONCLUSION: In this series, topical 5-aminosalicylic acid appears safe, effective, and well tolerated in the management of pregnant patients with distal colitis. PMID- 9399753 TI - Methotrexate in chronic active Crohn's disease: a double-blind, randomized, Israeli multicenter trial. AB - BACKGROUND: At present only one large controlled study has indicated that parenteral methotrexate may be effective in chronic active Crohn's disease (CD). AIM: To evaluate the effectiveness of oral methotrexate in chronic steroid dependent CD. PATIENTS: Patients with active CD, who have received steroids and/or immunosuppressives for at least 4 months during the preceding 12 months and with a current Harvey-Bradshaw index of > or = 7 were studied. METHODS: Methotrexate (12.5 mg weekly) or 6-mercaptopurine (50 mg daily), or placebo were given during the 9 months of the trial in addition to steroids and 5 aminosalicylic acid as clinically indicated. RESULTS: Eighty-four patients were included (methotrexate, 26 patients; 6-mercaptopurine, 32 patients; placebo, 26 patients). The proportion of patients entering first remission as well as the proportions of patients relapsing after first remission were not significantly different between the groups. The mean Harvey-Bradshaw index and the mean monthly steroid dose were also similar. However, when each patient was evaluated as his or her own control, the reduction in steroid dose, the general well being, and the reduction in abdominal pain were significantly better in the methotrexate treated patients. CONCLUSIONS: Methotrexate at a weekly oral dose of 12.5 mg was found to be moderately better than 6-mercaptopurine and placebo in patients with chronic active CD. PMID- 9399754 TI - High prevalence of celiac disease among patients with insulin-dependent (type I) diabetes mellitus. AB - OBJECTIVES: Diagnosis of unrecognized celiac disease is potentially important. The prevalence of celiac disease in patients with insulin-dependent diabetes mellitus is uncertain. We report the prevalence of celiac disease in a stratified random sample (n = 101) of adult insulin-dependent diabetic patients (age, 18-59 yr) attending our clinic, and in an age- and sex-matched control group (n = 51). METHODS: Screening was by anti-endomysial antibody, measured by indirect immunofluorescence using sections of human umbilical cord. RESULTS: Celiac disease had not been suspected in any patient at the time of screening. Eight patients tested positive for anti-endomysial antibody, all of whom had a distal duodenal biopsy performed. Five patients had histologic evidence of celiac disease. One patient with negative histology was receiving immunosuppressive therapy for a renal-pancreas transplant. Of the five patients with abnormal histology, two improved on gluten restriction, one was unable to comply, one refused treatment, and one was lost to follow-up. No control subject tested positive for endomysial antibody. CONCLUSIONS: Patients with insulin-dependent diabetes have an increased prevalence of celiac disease. Because most cases are clinically unrecognized, consideration should be given to screening all insulin dependent diabetes mellitus patients with endomysial antibodies. PMID- 9399755 TI - Randomized comparison of ranitidine bismuth citrate-based triple therapies for Helicobacter pylori. AB - OBJECTIVES: In an attempt to increase the efficacy and simplicity of FDA-approved regimens for Helicobacter pylori, we studied (1) addition of an inexpensive antibiotic (amoxicillin) to twice-daily ranitidine bismuth citrate (RBC) clarithromycin dual therapy, and (2) substitution of RBC for bismuth subsalicylate + H2-receptor antagonist in bismuth-based triple therapy. METHODS: Subjects with previously untreated Helicobacter pylori infection documented by 13C-urea breath test plus either endoscopic biopsy or serology were randomly assigned to a 2-wk course of (1) RBC 400 mg b.i.d., amoxicillin 1 g b.i.d., and clarithromycin 500 mg b.i.d. (RAC), or (2) RBC 400 mg b.i.d., metronidazole 250 mg t.i.d., and tetracycline 500 mg t.i.d. (RMT). Repeat breath test was performed 4 wk after the completion of therapy. RESULTS: Intent-to-treat and per-protocol cure rates for RAC were 46 of 50 patients (92%) and 45 of 47 patients (96%); for RMT they were 40 of 50 patients (80%) and 37 of 42 patients (88%). Study drugs were stopped due to side effects in three patients (6%) taking RAC and six patients (12%) taking RMT. CONCLUSIONS: Twice-daily RBC-based triple therapy with clarithromycin and amoxicillin produces Helicobacter pylori eradication rates over 90%, which is comparable to rates seen with proton pump inhibitor-based triple therapies. RBC also may be substituted for bismuth subsalicylate and an + H2-receptor antagonist in standard bismuth-based triple therapy. PMID- 9399756 TI - Is duodenal gastric metaplasia a consequence of Helicobacter pylori infection in children? AB - BACKGROUND: Duodenal gastric metaplasia (DGM) is commonly found in association with Helicobacter pylori (Hp)-associated gastritis in adults. DGM is also considered a risk factor for duodenal ulcer development. The prevalence of DGM in children and its association with gastritis, duodenitis, or the presence of Hp organisms is not clear. We investigated the prevalence of DGM in children and explore its association with several possible risk factors, including age, gender, gastritis, duodenitis, or Hp presence in the gastric antrum. METHODS: A retrospective analysis of 173 upper endoscopy procedures performed between 1993 and 1995 at Cabell Huntington Hospital, Huntington, WV, was done. Gastric and duodenal biopsies were stained with Giemsa for Hp detection, periodic acid-Schiff for DGM, and hematoxylin and eosin for histologic assessment. Gastric mucosal inflammation was graded according to Sydney criteria. RESULTS: Duodenal gastric metaplasia was identified in 23 of 173 (13%) patients. Duodenitis but not age, gender, gastritis, or the presence of Hp in the gastric antrum was associated with DGM development. In 4 of 23 DGM foci, Hp was identified. CONCLUSIONS: In children, DGM is not the consequence of Hp infection. PMID- 9399757 TI - Prevalence and pattern of Helicobacter pylori gastritis in the gastric cardia. AB - OBJECTIVES: Helicobacter pylori has a predilection for antral colonization. Local acid production is the major determinant of colonization. Because production is low in the antrum and cardia, H. pylori should also colonize the cardia. We therefore investigated the histologic pattern of gastritis and the prevalence of H. pylori in the cardia compared with the antrum and corpus. METHODS: From 135 H. pylori-infected patients with gastritis, ulcer disease, or reflux esophagitis, biopsies were obtained from the antrum, corpus, and cardia. The prevalence, topography, and histologic parameters of gastritis were examined. RESULTS: All 135 patients had active antral H. pylori gastritis: in the cardia, 132 of these patients (97.7%) showed active gastritis, and 124 patients (91.9%) had H. pylori visible on staining. Gastritis of the cardia in most patients resembled antral gastritis, but the density of bacteria and the inflammatory responses were less marked. The most striking finding in the cardia of patients with gastroesophageal reflux was a lower density of bacteria compared with antrum and corpus. Intestinal metaplasia was found in 32 patients in antral mucosa (23.7%) versus 28 patients in the cardia (20.7%), versus 11 patients in the corpus (8.1%), and was multifocal in 17 patients (12.6%). CONCLUSIONS: H. pylori gastritis commonly involves the cardia. The histologic density of the bacteria and inflammatory responses are lower than in the antrum. Intestinal metaplasia in the cardia is a common finding in H. pylori gastritis. The cause of the lower bacterial density in the cardia of patients with reflux esophagitis needs further investigation. PMID- 9399758 TI - Usefulness of anti-Helicobacter pylori and anti-CagA antibodies in the selection of patients for gastroscopy. AB - OBJECTIVES: Screening of dyspeptic patients with serological tests for Helicobacter pylori before open-access gastroscopy has been suggested to be worthwhile. CagA-positive H. pylori strains may be associated with major pathology more often than CagA-negative strains. The usefulness of anti-H. pylori and anti-CagA antibodies in screening for gastroscopy was evaluated in unselected dyspeptic patients. METHODS: Four hundred consecutive, unselected dyspeptic patients (mean age, 56.8 yr) in primary care were investigated with gastroscopy, ultrasonography of the upper abdomen, laboratory tests (including serological tests for H. pylori and CagA), and other examinations if needed. The patients were followed for 1 yr. RESULTS: Results of serological tests were positive for H. pylori in 56.2% of patients, of whom 64.4% also had results positive for CagA. Use of H. pylori and CagA serology-based screening combined with a history of nonsteroidal anti-inflammatory drug use would have detected only 80 and 70% of the major pathologies (peptic ulcer, moderate or severe esophagitis, celiac disease, or malignancy), respectively, in these patients. Gastroscopy would have been avoided in 30 and 41%, respectively, if only patients who had positive results on serological tests or who were nonsteroidal anti-inflammatory drug users would have been referred. In patients younger than 45 yr of age (n = 87), 60-74% of gastroscopies would have been avoided, but 50-60% of major pathologies would have been missed, by using the screening strategy studied. One of the nine malignancies (all in patients >45 yr of age) was H. pylori-negative, and two were CagA-negative. CONCLUSIONS: Anti-CagA antibodies do not offer advantages compared with anti-H. pylori antibodies in screening patients for gastroscopy. A remarkable share of major pathologies are missed by both of these screening methods. Therefore, the results of these screening tests are not recommended as selective criteria for gastroscopy. PMID- 9399759 TI - Storage temperature of the unbuffered rapid urease test. AB - OBJECTIVES: The unbuffered rapid urease test (RUT) is an accurate, rapid, and inexpensive method for detecting Helicobacter pylori. However, it is generally recommended that the reagent be prepared daily. This prospective study was undertaken to evaluate the shelf life of our unbuffered RUT when stored at 4 and 20 degrees C. METHODS: Ninety-five patients were studied. Three sets of antral (X2) and body (X1) biopsy samples were taken from each patient. The samples were subjected to histological examination, with the RUTs stored at 4 and -20 degrees C. The RUT tubes were examined at 1 and 15 min. RESULTS: Fifty-six patients (59%) were infected with H. pylori as defined by histological examination. The reagent was classified according to storage time (group I, < or = 5 days; group II, > 5 days). The mean (SD) storage time of group I (n = 59) and group II (n = 36) was 3.2 (1.4) and 9.9 (5.0) days, respectively. At 15 min, the sensitivity of our RUT stored at 4 degrees C was significantly higher in group I than in group II (92 vs 47%). On the other hand, the sensitivity of our RUT stored at -20 degrees C remained consistently high in both groups (15 min: group I, 92%; group II, 100%). Our RUTs stored at 4 and -20 degrees C were highly specific in both groups. CONCLUSIONS: Our RUT remains highly sensitive and specific when it is stored at 4 degrees C for up to 5 days. When the RUT is expected to be stored for a longer period of time, the bottles should be frozen at -20 degrees C. PMID- 9399760 TI - Determination of hepatitis delta virus (HDV)-RNA in asymptomatic cases of HDV infection. AB - OBJECTIVES: To assess the frequency of hepatitis delta virus (HDV) viremia in asymptomatic cases of HDV infection and the clinical significance of the HDV viremia, we conducted a cross-sectional, community-based study. METHODS: Of 2207 examinees, 210 (9.5%) were found to be positive for hepatitis B surface antigen (HBsAg). Antibody to HDV was detected in 47 (22.4%) of the 210 examinees, and 43 of the 47 were further evaluated for serum HDV-RNA by polymerase chain reaction. RESULTS: Twenty-one (48.8%) of the 43 had detectable levels of HDV-RNA in serum, and 22 (51.2%) were negative for serum HDV-RNA. The majority (61.9%) of the HDV RNA-positive HBsAg carriers had high levels of serum ALT. In contrast, the frequency of an abnormally high level of serum ALT was only 9.1% in the HBsAg carriers positive for HDV antibody but negative for HDV-RNA, and the frequency did not differ from that seen in the HBsAg-negative individuals. The semiquantified HDV-RNA levels did not correlate with the serum ALT levels. CONCLUSION: Seropositivity of HDV-RNA was strongly associated with liver cell damage, even in asymptomatic cases. The absence of a detectable level of serum HDV-RNA might be related to previous HDV infection. PMID- 9399761 TI - Idiopathic colonic inflammation in AIDS: an open trial of prednisone. AB - OBJECTIVES: Idiopathic colonic inflammation and ulceration have been described in HIV infection, but only as isolated case reports. We have been treating this condition with a uniform corticosteroid protocol and now report our results. METHODS: We describe the cases of eight patients with HIV infection who had diarrhea for more than 4 wk and inflammation and/or ulceration in the colon at endoscopy, confirmed by biopsy, without any invasive pathogens despite extensive evaluation. Each patient was treated with prednisone, starting at 40 mg/day, then tapered according to a standardized protocol. RESULTS: The diarrhea completely resolved in three patients and partially improved in five. One patient had some improvement but was unable to tolerate the prednisone because of a severe exacerbation of anal warts. He responded to subtotal colectomy. After a minimum follow-up of 8 months (mean, 17 months), only one patient (complete response to prednisone) was found to have an enteric pathogen. In this patient, cytomegalovirus colitis was diagnosed 15 months after prednisone was started. CONCLUSION: Idiopathic colonic inflammation or ulceration in HIV infection (1) may respond to corticosteroid therapy without life-threatening side effects and (2) is only rarely followed by the detection of a recognized pathogen. These observations suggest that enteric pathogens are not missed by standard techniques. PMID- 9399762 TI - Differences in risk of Crohn's disease in offspring of mothers and fathers with inflammatory bowel disease. AB - OBJECTIVE: To determine whether there are any unusual patterns of transmission of susceptibility to inflammatory bowel disease (IBD) within multiplex families. METHODS: Individuals with IBD were recruited for genome-wide screening of susceptibility genes. The extent of familial aggregation and blood relationships in multiplex families were determined by questionnaires given to participants followed up by confirmation of disease diagnosis by participants' physicians. RESULTS: Of 135 families identified in which both a parent and a child had IBD, 93 involved transmission of susceptibility to disease from mother to child versus 42 examples of transmission from father to child (p = 0.00001, exact two-tailed binomial test). This distortion in transmission on the basis of the sex of the parent was observed only among non-Jewish pairs with Crohn's disease (CD), in which, of 33 parent-child pairs with CD, disease susceptibility was transmitted from the mother 28 times (p = 0.00007). CONCLUSION: Susceptibility to CD in a subset of patients may involve a gene that is imprinted. PMID- 9399763 TI - Increased portal tract infiltration of mast cells and eosinophils in primary biliary cirrhosis. AB - OBJECTIVES: Although recent reports demonstrate that eosinophilia is a distinctive feature of early stage primary biliary cirrhosis (PBC), the pathogenesis of this eosinophilia remains unknown. Clinical and experimental data suggest potential mast cell activation in cholestatic liver diseases. Because mast cell-derived mediators could induce eosinophil chemotaxis and activation, we hypothesized that mast cell activation may play a role in the pathogenesis of eosinophilia in PBC. Thus, as the first step in examining a possible link between mast cell activation and eosinophilia in PBC, we quantified the infiltration of mast cells and eosinophils in the livers of patients with PBC. METHODS: The study population consisted of 11 patients with stage I or stage II PBC and 11 patients with chronic viral hepatitis. Mast cells and eosinophils were identified by immunohistochemistry using monoclonal antibodies against mast cell tryptase (AA1) and eosinophilic cationic protein (EG2), respectively. Cell infiltration in the portal tract was quantitated morphometrically. RESULTS: When compared with patients with chronic viral hepatitis, patients with PBC showed significantly increased portal infiltration with mast cells (140 +/- 25 vs 72 +/- 10 cells/mm2 [mean +/- SEM, p < 0.05]) and eosinophils (76 +/- 19 vs 32 +/- 9 cells/mm2 [p < 0.05]). Numbers of portal mast cells correlated with numbers of eosinophils in patients with PBC (r = 0.60, p < 0.05) but not in patients with chronic hepatitis (r = 0.34, p = 0.47). CONCLUSION: Concomitant increases in mast cell and eosinophil infiltration in the portal tract suggest a role for mast cell activation in the pathogenesis of eosinophilia in PBC. PMID- 9399764 TI - Octreotide inhibition of flushing and colonic motor dysfunction in carcinoid syndrome. AB - OBJECTIVES: Previous studies showed increased plasma motilin and substance P concentrations and accelerated motor function in the small bowel and colon in patients with carcinoid diarrhea. Octreotide is beneficial in patients with carcinoid syndrome. Our hypothesis was that octreotide inhibits accelerated motility and gut neuropeptides in carcinoid syndrome. METHODS: In 12 patients with metastatic carcinoid syndrome, we investigated the effect of octreotide 50 microg s.c. t.i.d (n = 6) or placebo (n = 6) on postprandial symptoms, GI transit, colonic motility, and circulating levels of selected circulating peptides and amines. RESULTS: Octreotide reduced postprandial flushing (p = 0.03) but not pain. Octreotide significantly retarded overall colonic transit and proximal colonic emptying (p < 0.05); it tended to prolong small bowel transit time (p = 0.13) and to reduce postprandial colonic tone (p = 0.08) compared with placebo. Octreotide also reduced circulating levels of peptide YY, neurotensin, vasoactive intestinal polypeptide, and substance P but had no effect on plasma motilin, neuropeptide Y, calcitonin gene-related peptide, or histamine after meal ingestion. CONCLUSION: Octreotide ameliorates gut motor dysfunctions that characterize carcinoid diarrhea; the potential role of specific antagonism of serotonin, substance P, and vasoactive intestinal polypeptide alone or in combination with agents that inhibit their release in carcinoid diarrhea deserves further study. PMID- 9399765 TI - Clinical and microbiological features of liver abscess after transarterial embolization for hepatocellular carcinoma. AB - OBJECTIVES: To present the clinical and microbiological features of liver abscess after transarterial embolization (TAE) for hepatocellular carcinoma (HCC). METHODS: We retrospectively reviewed records of 452 TAE procedures in 289 patients with HCC over a 2-yr period. RESULTS: Four men and one woman with a mean age of 68.4 yr were diagnosed with liver abscess 1-8 wk (mean 4.6 wk) after the embolization. The incidence was 1.1% (5/452). Common symptoms included fever, chills, and right upper quadrant pain. Serum aminotransferase, alkaline phosphatase, and gamma-glutamyltransferase levels and leukocyte count were frequently elevated. All the abscesses appeared as areas of hypodensity on CT scan and hypoechogenicity on ultrasonogram. The areas contained gas in the embolized tumor, which led to the suspicion and finally the diagnosis of abscess. In contrast to predominance of gram-negative aerobes in sporadic pyogenic liver abscesses, the causative microorganism was predominantly gram positive (60%). All patients were treated with parenteral antibiotics plus percutaneous aspiration, drainage, or operation, but one patient died from the abscess. CONCLUSIONS: For patients receiving TAE for HCC, few specific clinical or radiological features could readily differentiate patients complicated with liver abscess from those without. This may delay a timely diagnosis and lead to significant morbidity. Hence, in patients with risk factors, including old age, previous biliary tract disease, large tumor size (>5 cm), and gas forming in the embolized tumor, aspiration of the suspected focal hepatic lesion should be performed as soon as possible. PMID- 9399766 TI - Zinc, copper, and superoxide dismutase in hepatocellular carcinoma. AB - Superoxide dismutase (Cu/Zn), a metalloenzyme, activity was found to be significantly lower in the human hepatocellular carcinoma (hepatoma tissue) (0.78 +/- 0.33 U/microg protein) compared with that seen in the surrounding "normal" or cirrhotic tissue (1.40 +/- 0.48 U/microg; 1.27 +/- 0.64 U/microg). The SOD activity of "normal" appearing liver cells adjacent to the tumor or cirrhotic tissue is still lower than that level observed in the normal liver from the subjects without known liver disease (1.40 +/- 0.48 U/microg vs 2.94 +/- 1.53 U/microg). We have also observed that the trace elements of zinc and copper, which are essential in expressing the enzyme activity are also significantly lower in the hepatoma (22.54 +/- 6.73 microg and 2.83 +/- 1.16 microg per gram of tissue) compared with those in the surrounding "normal" hepatic tissue (Zn2+, 64.36 +/- 9.17 microg/g; Cu2+, 11.43 +/- 4.74 microg/g). The difference in the zinc content between the "normal" and the cirrhotic tissue is also significant (64.36 +/- 9.17 microg/g vs 42.37 +/- 10.97 microg/g). However, the copper content in the cirrhotic tissue is higher but not statistically different from that level in the "normal" tissue (15.53 +/- 5.90 microg/g vs 11.43 +/- 4.74 microg/g). Furthermore, the plasma zinc level is significantly lower among the patients who have suffered from hepatoma compared with those subjects without known liver disease (631.73 +/- 52.43 microg/L vs 845.53 +/- 68.13 microg/L). Our data suggest that the superoxide dismutase activity is impaired in the hepatoma tissue. The lower concentration of trace elements (Zn2+ and Cu2+) found in the hepatoma tissue may contribute to cause the difference in the observed enzymic activities. PMID- 9399767 TI - Pancreatic-biliary ascariasis: experience of 300 cases. AB - BACKGROUND: Infestation with Ascaris lumbricoides is seen worldwide. Recently, there has been much interest in the pancreatic-biliary complications of Ascaris infection. In this study, we present our experience of 300 patients seen in a tertiary referral center. MATERIALS AND METHODS: Case charts of patients seen in the Department of Gastroenterology, University of Damascus, Syria, were analyzed, retrospectively, over a 5-yr period (September of 1988 to August of 1993). During this period, 1666 endoscopic retrograde cholangiopancreatographic studies were performed and pancreatic-biliary ascariasis was diagnosed in 300 patients (18%). RESULTS: The most common presenting symptom was abdominal pain, seen in 98% of patients (294 patients). Complications observed were ascending cholangitis (48 patients; 16%), acute pancreatitis (13 patients; 4.3%), and obstructive jaundice (4 patients; 1.3%). History of worm emesis was present in 25% of patients (76 patients). Most patients (240 patients; 80%) had previously undergone a cholecystectomy or an endoscopic sphincterotomy (232 patients; 77%). Worms were successfully extracted endoscopically in all except two patients, and there were no procedure-related complications. CONCLUSIONS: In endemic countries, ascariasis should be suspected in patients with pancreatic-biliary disease, especially if a cholecystectomy or sphincterotomy has been performed in the past. Endoscopic management results in rapid resolution of symptoms and prevents development of complications. PMID- 9399768 TI - Monoethylglycinexylidide formation measurement as a hepatic function test to assess severity of chronic liver disease. AB - OBJECTIVES: Monoethylglycinexylidide (MEGX) is the main lidocaine metabolite and its formation depends on liver microsomal activity. MEGX formation was studied in comparison with the histological score of chronic hepatitis and with the clinical score (Child-Pugh) of cirrhosis. Furthermore, we evaluated its ability to distinguish between the two liver diseases. METHODS: We studied 284 patients: 130 with chronic hepatitis (on the basis of the histological activity index, 45 had mild chronic hepatitis, 54 had moderate chronic hepatitis, and 31 had chronic hepatitis with cirrhosis) and 154 with cirrhosis (49 Child-Pugh's class A, 78 class B, and 27 class C). MEGX formation was evaluated 15, 30, and 60 min after lidocaine administration. RESULTS: MEGX formation showed a stepwise decline corresponding to worsened liver disease. MEGX values were related both to the histological score in chronic hepatitis and to the clinical score in cirrhosis. Significantly lower values were found in females < 50 yr of age than in males of the same age. The MEGX test showed great efficacy in discriminating between chronic hepatitis and cirrhosis compared with standard liver tests. CONCLUSIONS: Measurement of MEGX formation proved to be a safe test, allowing us to show that functional subgroups can be identified both in chronic hepatitis and in cirrhosis. Thus, this test could integrate both the histological grading of chronic hepatitis and the clinical staging of cirrhosis. PMID- 9399769 TI - Gallbladder motility and cholecystokinin secretion during continuous enteral nutrition. AB - OBJECTIVES: During total parenteral nutrition, gallbladder motility is impaired, resulting in sludge and stone formation. Little is known about gallbladder motility during prolonged enteral nutrition. METHODS: We studied gallbladder motility during continuous enteral nutrition (CEN) in nine hospitalized patients with active inflammatory bowel disease. The patients received a polymeric diet (2000 kcal/24 h) by CEN through a nasogastric tube for a prolonged period. Gallbladder volumes were obtained daily by ultrasonography, starting from day 0 (before CEN) and on 7 consecutive days during CEN. At days 0, 1, 4, and 7, the gallbladder response to i.v. cholecystokinin (CCK-33; 0.5 Ivy Dog unit/kg/h) was studied. Plasma CCK levels were determined at regular intervals by radioimmunoassay. RESULTS: No significant differences were observed on day 0 between patients and a group of nine healthy control subjects in fasting gallbladder volumes (19.4 +/- 2.3 and 19.6 +/- 2.4 cm3, respectively) and gallbladder contraction during CCK infusion (56 +/- 14% and 69 +/- 7%, respectively). During CEN, from day 1 to day 7, mean gallbladder volume remained significantly (p < 0.05) reduced compared with fasting gallbladder volume, and mean plasma CCK levels remained significantly (p < 0.05) increased compared with fasting levels. Although gallbladder volume was significantly reduced during CEN, the gallbladder contractile response to CCK was not affected; at days 1, 4, and 7, gallbladder contraction was 36-57%. CONCLUSIONS: During CEN, 1) gallbladder volume is significantly reduced and plasma CCK levels are significantly increased, 2) these effects are sustained over time (7 days), and 3) the gallbladder remains responsive to exogenous CCK. These results indicate that gallbladder contractility and gallbladder responsiveness to CCK are preserved during prolonged CEN in patients with inflammatory bowel disease. PMID- 9399770 TI - Acid steatocrit: a simple, rapid gravimetric method to determine steatorrhea. AB - OBJECTIVES: The detection and evaluation of steatorrhea in a rapid, quantitative fashion are clinically needed in patients with suspected steatorrhea. Our aim was to evaluate the acid steatocrit method, on random spot stools in adults with and without steatorrhea, relative to the qualitative (microscopic) and quantitative assessments for fecal fat. METHODS: Stool samples were collected 72 h after a diet of 100 g of fat per day and randomly from 15 healthy controls, 14 patients with chronic pancreatitis, and seven patients with small bowel disease. All stools had quantitative, qualitative, and acid steatocrit analyses performed for fecal fat. RESULTS: The sensitivity and specificity for the detection of steatorrhea by the spot stool qualitative fecal fat were 78 and 70%, respectively. The spot stool acid steatocrit correlated linearly with the 72-h stool quantitative fecal fat (g/24 h), r = 0.761 and p < 0.001. The acid steatocrit on random spot stools, compared with the 72-h stool quantitative fecal fat, revealed a sensitivity of 100%, a specificity of 95%, and a positive predictive value of 90% for the detection of steatorrhea. It also estimated the quantitative fecal fat. CONCLUSIONS: The acid steatocrit can be performed accurately on random spot stools and can be used to detect the presence of steatorrhea and estimate the quantitative fecal fat. This assay can be done with readily available equipment for rapid evaluation. Use of a spot stool sample simplifies the acid steatocrit, further improving on the practicality of this test. This study also confirms the clinical usefulness of this simplified method to detect steatorrhea. PMID- 9399771 TI - Inflammatory bowel disease and immune thrombocytopenic purpura: is there a correlation? AB - Different hematologic abnormalities are often encountered in patients with inflammatory bowel disease. Among them anemia, leukocytosis, and thrombocytosis are commonly seen. Leukopenia and thrombocytopenia are observed mostly as a side effect of therapy, particularly with use of immunosuppressive drugs. Immune thrombocytopenic purpura is rarely reported in association with inflammatory bowel disease. We present two cases with combination of these entities along with a literature review and treatment options. Immune thrombocytopenic purpura in these patients presented as an extraintestinal manifestation of inflammatory bowel disease mediated by a disturbance of the immune system. PMID- 9399772 TI - Dietl's crisis: a syndrome of episodic abdominal pain of urologic origin that may present to a gastroenterologist. AB - A 53-yr-old woman presented with a 2-yr history of recurrent episodes of severe abdominal pain and nausea. Multiple investigations by a general surgeon, a urologist, and a gastroenterologist failed to identify the cause. She was referred to our Biliary Service for ERCP and sphincter of Oddi manometry. However, a detailed history was inconsistent with biliary pain, and the patient, having discussed the risks and benefits, elected not to proceed with ERCP. The patient was asked to come to the hospital during an acute attack of her pain for assessment. When this was done, transabdominal ultrasound revealed right hydronephrosis; intravenous urography showed obstruction at the level of the ureteropelvic junction, consistent with the presence of an aberrant artery. The syndrome of episodic abdominal pain and hydronephrosis caused by extrinsic pressure from such an artery is known as Dietl's crisis. In our patient, the diagnosis was confirmed at surgery, when the ureteric obstruction was dealt with by pyeloplasty. She made an uneventful recovery and remains asymptomatic 12 months later. The keys to diagnosing Dietl's crisis are awareness of the entity, taking a detailed pain history, and timely cross-sectional abdominal imaging during an attack. PMID- 9399773 TI - Gastric antral vascular ectasia with portal hypertension: treatment with TIPSS. AB - Gastric antral vascular ectasia, or "watermelon stomach," is a distinct clinical entity within the spectrum of upper gastrointestinal mucosal vascular abnormalities. The pathobiology of the disease is unclear. We here describe a case that emphasizes the importance of the "watermelon stomach" as differential diagnosis of occult gastrointestinal blood loss and report on its successful treatment through installation of a transjugular intrahepatic portosystemic stent shunt (TIPSS). PMID- 9399774 TI - Management of ingested foreign bodies within the appendix: a case report with review of the literature. AB - A case is reported of an elective appendectomy in a patient with known ingestion of a sharp foreign body. The metal drill bit was ingested unintentionally 3 months before presentation at our institution. Plain abdominal films demonstrated the foreign body in the right lower abdominal quadrant. Because the gold dental drill bit was sharp and thought to be lodged in the terminal ileum or cecum, an attempt was made to remove the object during colonoscopy. This attempt was unsuccessful because no drill bit could be detected in the colon or terminal ileum. A laparoscopic exploration was performed, and the foreign body was found to lie in the appendix, after bowel manipulation under fluoroscopic guidance and with direct laparoscopic visualization. A laparoscopic assisted appendectomy was performed. On pathologic examination the drill bit was embedded in the tip of the appendix with signs of intramucosal acute inflammation. Management and indication for surgery of foreign bodies in the appendix are discussed, and we review the related literature. This is the second reported case of a dental drill bit in the appendix causing appendicitis. PMID- 9399775 TI - Malignant lymphoma of the transverse colon associated with macroglobulinemia. AB - We present an unusual case of a malignant lymphoma of the transverse colon associated with macroglobulinemia. A 73-yr-old man was incidentally discovered to have high serum gamma-globulin on a regular check-up. Serum immunoquantitation revealed an IgM level of 3490 mg/dl. Kappa-type Bence-Jones protein was positive in the urine. Immunoelectrophoresis identified the abnormal protein as IgM-kappa. After hospitalization an abdominal tumor was detected with barium and CT, identified as a tumor of the transverse colon. Partial resection of the transverse colon was carried out. Histopathologically the tumor were confirmed as small lymphocytic non-Hodgkin lymphoma of B-cell origin, based on the Working Formulation. According to flowcytometric analysis, the tumor cells were positive for IgM-kappa. The lymphoma cells produced monoclonal IgM, giving rise to macroglobulinemia. PMID- 9399776 TI - Acute pancreatitis after long-term 5-aminosalicylic acid therapy. AB - Acute pancreatitis is a known, although rare, complication of mesalamine treatment. This complication typically appears within the first days or weeks after initiation of therapy. We describe two cases of acute pancreatitis that occurred after long term mesalamine therapy for ulcerative colitis. A rechallenge, performed in both patients, confirmed the diagnosis of mesalamine induced pancreatitis. These case reports provide evidence that 5-aminosalicylic acid may induce acute pancreatitis after long term treatment. PMID- 9399777 TI - Synchronous adenomas in a colonic interposition graft and the native colon. PMID- 9399778 TI - Use of transjugular intrahepatic portosystemic shunt as a bridge to transplantation in fulminant hepatic failure due to Budd-Chiari syndrome. AB - We report a case of fulminant hepatic failure in a 55-yr-old man due to Budd Chiari syndrome in the setting of polycythemia rubra vera. The patient presented with acute hepatic failure, which rapidly progressed to grade IV hepatic encephalopathy. Placement of a transjugular intrahepatic portosystemic shunt resulted in marked improvement of the encephalopathy and stabilized the liver failure. Subsequently, he underwent successful nonemergent orthotopic liver transplantation. Transjugular intrahepatic portosystemic shunt placement is a safe, effective, therapeutic option to bridge patients with fulminant Budd-Chiari to liver transplantation. PMID- 9399780 TI - Ulcerative colitis, hyperamylasemia, and asymptomatic pancreatic calcifications: making the case for pancreatitis as an extra luminal manifestation. PMID- 9399779 TI - Pneumomediastinum during relapse of ulcerative colitis. AB - Pneumomediastinum can be caused by gas dissecting along fascial tissue planes into the mediastinum from remote locations, including the retroperitoneum. One potential source of retroperitoneal gas is the colon. We present the third reported case of pneumomediastinum (plus pneumothorax and subcutaneous emphysema) without free intraperitoneal gas developing during an attack of ulcerative colitis. Because there was no colonic perforation noted at colectomy, the extracolonic gas was presumed to originate from a microscopic or partial thickness perforation of the colon. GI perforation must be considered not only in patients who have free intraperitoneal gas but also in those who present with symptoms, signs, or studies consistent with retroperitoneal gas, such as subcutaneous emphysema, pneumomediastinum, or pneumothorax. PMID- 9399782 TI - Factor V Leiden mutation presenting as repeated massive GI hemorrhages secondary to venous thrombosis. PMID- 9399781 TI - Mirizzi's syndrome with a high CA19-9 level mimicking cholangiocarcinoma. PMID- 9399783 TI - Thalidomide for AIDS diarrhea? PMID- 9399784 TI - Cure of Helicobacter pylori: a hidden curse? PMID- 9399785 TI - Why do we need a newer generation of recombinant vaccines against hepatitis B? PMID- 9399786 TI - Acid control and regression of Barrett's esophagus: is the glass half full or half empty? PMID- 9399787 TI - Oxandrolone use in Crohn's disease. PMID- 9399788 TI - Placement of a percutaneous gastrostomy tube in patients with an existing esophageal prosthesis. PMID- 9399789 TI - Enhanced expression of CA19-9 in mucinous gastric carcinoma. PMID- 9399790 TI - GenBank. AB - The GenBank(R) sequence database (http://www.ncbi.nlm.nih.gov/) incorporates DNA sequences from all available public sources, primarily through the direct submission of sequence data from individual laboratories and from large-scale sequencing projects. Most submitters use the BankIt (WWW) or Sequin programs to send their sequence data. Data exchange with the EMBL Data Library and the DNA Data Bank of Japan helps ensure comprehensive worldwide coverage. GenBank data is accessible through NCBI's integrated retrieval system, Entrez , which integrates data from the major DNA and protein sequence databases along with taxonomy, genome and protein structure information. MEDLINE(R) abstracts from published articles describing the sequences are also included as an additional source of biological annotation. Sequence similarity searching is offered through the BLAST series of database search programs. In addition to FTP, e-mail and server/client versions of Entrez and BLAST, NCBI offers a wide range of World Wide Web retrieval and analysis services of interest to biologists. PMID- 9399791 TI - The EMBL nucleotide sequence database. AB - The EMBL Nucleotide Sequence Database (http://www.ebi.ac.uk/embl. html ) constitutes Europe's primary nucleotide sequence resource. DNA and RNA sequences are directly submitted from researchers and genome sequencing groups and collected from the scientific literature and patent applications (Fig. 1). In collaboration with DDBJ and GenBank the database is produced, maintained and distributed at the European Bioinformatics Institute. Database releases are produced quarterly and are distributed on CD-ROM. EBI's network services allow access to the most up-to-date data collection via Internet and World Wide Web interface, providing database searching and sequence similarity facilities plus access to a large number of additional databases. PMID- 9399792 TI - DNA Data Bank of Japan at work on genome sequence data. AB - We at the DNA Data Bank of Japan (DDBJ) (http://www.ddbj.nig.ac.jp) have recently begun receiving, processing and releasing EST and genome sequence data submitted by various Japanese genome projects. The data include those for human, Arabidopsis thaliana, rice, nematode, Synechocystis sp. and Escherichia coli. Since the quantity of data is very large, we organized teams to conduct preliminary discussions with project teams about data submission and handling for release to the public. We also developed a mass submission tool to cope with a large quantity of data. In addition, to provide genome data on WWW, we developed a genome information system using Java. This system (http://mol.genes.nig.ac.jp/ecoli/) can in theory be used for any genome sequence data. These activities will facilitate processing of large quantities of EST and genome data. PMID- 9399793 TI - The Genome Sequence DataBase (GSDB): improving data quality and data access. AB - In 1997 the primary focus of the Genome Sequence DataBase (GSDB; www. ncgr.org/gsdb ) located at the National Center for Genome Resources was to improve data quality and accessibility. Efforts to increase the quality of data within the database included two major projects; one to identify and remove all vector contamination from sequences in the database and one to create premier sequence sets (including both alignments and discontiguous sequences). Data accessibility was improved during the course of the last year in several ways. First, a graphical database sequence viewer was made available to researchers. Second, an update process was implemented for the web-based query tool, Maestro. Third, a web-based tool, Excerpt, was developed to retrieve selected regions of any sequence in the database. And lastly, a GSDB flatfile that contains annotation unique to GSDB (e.g., sequence analysis and alignment data) was developed. Additionally, the GSDB web site provides a tool for the detection of matrix attachment regions (MARs), which can be used to identify regions of high coding potential. The ultimate goal of this work is to make GSDB a more useful resource for genomic comparison studies and gene level studies by improving data quality and by providing data access capabilities that are consistent with the needs of both types of studies. PMID- 9399794 TI - The PIR-International Protein Sequence Database. AB - From its origin the Protein Information Resource (http://www-nbrf. georgetown.edu/pir/) has supported research on evolution and computational biology by designing and compiling a comprehensive, quality controlled, and well organized protein sequence database. The database has been produced and updated on a regular schedule since 1984. Since 1988 it has been maintained collaboratively by the PIR-International, an association of data collection centers engaged in international cooperation for the development of this research resource during a period of explosive acquisition of new data. As of June 1997, essentially all sequence entries have been classified into families, allowing the efficient application of methods to propagate and standardize annotation among related sequences. The databases are available through the Internet by the World Wide Web and FTP, or on CD-ROM and magnetic media. PMID- 9399795 TI - MIPS: a database for protein sequences and complete genomes. AB - The MIPS group [Munich Information Center for Protein Sequences of the German National Center for Environment and Health (GSF)] at the Max-Planck-Institute for Biochemistry, Martinsried near Munich, Germany, is involved in a number of data collection activities, including a comprehensive database of the yeast genome, a database reflecting the progress in sequencing the Arabidopsis thaliana genome, the systematic analysis of other small genomes and the collection of protein sequence data within the framework of the PIR-International Protein Sequence Database (described elsewhere in this volume). Through its WWW server (http://www.mips.biochem.mpg.de ) MIPS provides access to a variety of generic databases, including a database of protein families as well as automatically generated data by the systematic application of sequence analysis algorithms. The yeast genome sequence and its related information was also compiled on CD-ROM to provide dynamic interactive access to the 16 chromosomes of the first eukaryotic genome unraveled. PMID- 9399796 TI - The SWISS-PROT protein sequence data bank and its supplement TrEMBL in 1998. AB - SWISS-PROT (http://www.expasy.ch/) is a curated protein sequence database which strives to provide a high level of annotations (such as the description of the function of a protein, its domains structure, post-translational modifications, variants, etc.), a minimal level of redundancy and high level of integration with other databases. Recent developments of the database include: an increase in the number and scope of model organisms; cross-references to two additional databases; a variety of new documentation files and improvements to TrEMBL, a computer annotated supplement to SWISS-PROT. TrEMBL consists of entries in SWISS PROT-like format derived from the translation of all coding sequences (CDS) in the EMBL nucleotide sequence database, except the CDS already included in SWISS PROT. PMID- 9399797 TI - Compilation of DNA sequences of Escherichia coli K12: description of the interactive databases ECD and ECDC. AB - We have compiled the DNA sequence data for Escherichia coli K12 available from the GenBank and EMBL data libraries and independently from the literature. We provide the most definitive version of the ECD Escherichia coli database now exclusively via the World Wide Web System (http://susi.bio.uni giessen.de/ecdc.html ). Our database encloses the completed genome sequence recently published by two competing groups and an assembled set of all elder sequences. The organisation of the database allows precise physical location of each individual gene or regulatory region, even taking into consideration discrepancies in nomenclature. The WWW program allows to the user to branch into the original EMBL and SWISS-PROT datafiles. A number of links to other WWW servers dealing with E. coli is provided. A FASTA and BLAST search may be performed online. Besides the WWW format a flat file version may be obtained via ftp. A number of discrepancies between the two systematic sequence determinations and/or the literature have not yet been resolved. However, our database may serve as a reference source for resolution and/or the assignment of strain difference. PMID- 9399798 TI - EcoCyc: Encyclopedia of Escherichia coli genes and metabolism. AB - The encyclopedia of Escherichia coli genes and metabolism (EcoCyc) is a database that combines information about the genome and the intermediary metabolism of E.coli. The database describes 3030 genes of E.coli , 695 enzymes encoded by a subset of these genes, 595 metabolic reactions that occur in E.coli, and the organization of these reactions into 123 metabolic pathways. The EcoCyc graphical user interface allows scientists to query and explore the EcoCyc database using visualization tools such as genomic-map browsers and automatic layouts of metabolic pathways. EcoCyc can be thought of as an electronic review article because of its copious references to the primary literature, and as a (qualitative) computational model of E.coli metabolism. EcoCyc is available at URL http://ecocyc.PangeaSystems.com/ecocyc/ PMID- 9399799 TI - Genes and proteins of Escherichia coli K-12. AB - GenProtEC is a database of Escherichia coli genes and their gene products, classified by type of function and physiological role and with citations to the literature for each. Also present are data on sequence similarities among E.coli proteins, representing groups of paralogous genes, with PAM values, percent identity of amino acids, length of alignment and percent aligned. GenProtEC can be accessed at the URL http://www.mbl.edu/html/ecoli.html PMID- 9399800 TI - RegulonDB: a database on transcriptional regulation in Escherichia coli. AB - RegulonDB is a DataBase that integrates biological knowledge of the mechanisms that regulate the transcription initiation in Escherichia coli , as well as knowledge on the organization of the genes and regulatory signals into operons in the chromosome. The operon is the basic structure used in RegulonDB to describe the elements and properties of transcriptional regulation. The current version contains information around some 500 regulation mechanisms, essentially for sigma 70 promoters. PMID- 9399801 TI - The non-redundant Bacillus subtilis (NRSub) database: update 1998. AB - The non-redundant Bacillus subtilis database (NRSub) has been developed in the context of the sequencing project devoted to this bacterium. As this project has reached completion, the whole genome is now available as a single contig. Thanks to the ACNUC database management system and its associated retrieval system Query_win, each functional region of the genome can be accessed individually. Extra annotations have been added such as accession numbers for the genes, locations on the genetic map, codon adaptation index values, as well as cross references with other collections. NRSub is distributed through anonymous FTP as a text file in EMBL format and as an ACNUC database. It is also possible to access NRSub through two dedicated World Wide Web servers located in France (http://acnuc. univ-lyon1.fr/nrsub/nrsub.html ) and in Japan (http://ddbjs4h.genes. nig.ac.jp/ ). PMID- 9399802 TI - CyanoBase, a www database containing the complete nucleotide sequence of the genome of Synechocystis sp. strain PCC6803. AB - CyanoBase (http://www.kazusa.or.jp/cyano/) is a database containing genomic information on the cyanobacterium Synechocystis sp. strain PCC6803. It furnishes an annotation to each of the 3168 protein genes deduced from the entire nucleotide sequence of this genome. Information on the genome can be directly accessed through three different menus: a clickable physical map of the genome, a gene classification list, and a keyword search menu, all of which are accessible from the main page of the database. The entry page for a gene annotation contains the following information: the location of the gene on the genome, the nucleotide and deduced amino acid sequence of the gene, the result of a similarity search, and the classification of the deduced gene product according to its function. This page has reverse-links to the local physical map and gene classification list so that relevant genes can be searched in terms of their location on the genome and their function. In addition, the main page of CyanoBase provides engines for similarity searches between a query sequence and the entire genome sequence and for keyword searches, in addition to numerous links to pages containing related information. PMID- 9399803 TI - The Yeast Protein Database (YPD): a curated proteome database for Saccharomyces cerevisiae. AB - The Yeast Protein Database (YPD) is a curated database for the proteome of Saccharomyces cerevisiae . It consists of approximately 6000 Yeast Protein Reports, one for each of the known or predicted yeast proteins. Each Yeast Protein Report is a one-page presentation of protein properties, annotation lines that summarize findings from the literature, and references. In the past year, the number of annotation lines has grown from 25 000 to approximately 35 000, and the number of articles curated has grown from approximately 3500 to >5000. Recently, new data types have been included in YPD: protein-protein interactions, genetic interactions, and regulators of gene expression. Finally, a new layer of information, the YPD Protein Minireviews, has recently been introduced. The Yeast Protein Database can be found on the Web at http://www.proteome.com/YPDhome. html PMID- 9399805 TI - AtDB, the Arabidopsis thaliana database, and graphical-web-display of progress by the Arabidopsis Genome Initiative. AB - AtDB, the Arabidopsis thaliana Database, has a primary role to provide public access to the collected genomic information for A. thaliana via the World Wide Web (URL: http://genome-www.stanford. edu/ ). AtDB presents interactive physical and genetics maps that are hyperlinked with detailed information about the clones and markers placed on these maps. A large literature collection on Arabidopsis , contact information on researchers worldwide, laboratory method manuals and other information useful to plant molecular biologists are also provided. This paper discusses the database-driven clickable displays that provide easy navigation within a variety of genomic maps, including those summarizing progress of the international Arabidopsis genomic sequencing effort, AGI (the Arabidopsis Genome Initiative). The interface uses client-side hyperlinked GIF-images that direct the user to detailed database-information. A new BLAST service is also described. This gives users access to the thousands of Arabidopsis BAC clone end-sequences and includes hyperlinked images summarizing the search results. The linking of genetic and physically mapped regions and their sequence into information for loci within that region is an ongoing goal for this project. PMID- 9399804 TI - SGD: Saccharomyces Genome Database. AB - The Saccharomyces Genome Database (SGD) provides Internet access to the complete Saccharomyces cerevisiae genomic sequence, its genes and their products, the phenotypes of its mutants, and the literature supporting these data. The amount of information and the number of features provided by SGD have increased greatly following the release of the S.cerevisiae genomic sequence, which is currently the only complete sequence of a eukaryotic genome. SGD aids researchers by providing not only basic information, but also tools such as sequence similarity searching that lead to detailed information about features of the genome and relationships between genes. SGD presents information using a variety of user friendly, dynamically created graphical displays illustrating physical, genetic and sequence feature maps. SGD can be accessed via the World Wide Web at http://genome-www.stanford.edu/Saccharomyces/ PMID- 9399806 TI - FlyBase: a Drosophila database. AB - FlyBase (http://flybase.bio.indiana.edu/) is a comprehensive database of genetic and molecular data concerning Drosophila . FlyBase is maintained as a relational database (in Sybase) and is made available as html documents and flat files. The scope of FlyBase includes: genes, alleles (with phenotypes), aberrations, transposons, pointers to sequence data, gene products, maps, clones, stock lists, Drosophila workers and bibliographic references. PMID- 9399807 TI - FlyNets and GIF-DB, two internet databases for molecular interactions in Drosophila melanogaster. AB - GIF-DB and FlyNets are two WWW databases describing molecular (protein-DNA, protein-RNA and protein-protein) interactions occuring in the fly Drosophila melanogaster (http://gifts.univ-mrs.fr/GIFTS_home_page.html ). GIF-DB is a specialised database which focuses on molecular interactions involved in the process of embryonic pattern formation, whereas FlyNets is a new and more general database, the long-term goal of which is to report on any published molecular interaction occuring in the fly. The information content of both databases is distributed in specific lines arranged into an EMBL- (or GenBank-) like format. These databases achieve a high level of integration with other databases such as FlyBase, EMBL, GenBank and SWISS-PROT through numerous hyperlinks. In addition, we also describe SOS-DGDB, a new collection of annotated Drosophila gene sequences, in which binding sites for regulatory proteins are directly visible on the DNA primary sequence and hyperlinked both to GIF-DB and TRANSFAC database entries. PMID- 9399809 TI - Virgil: a database of rich links between GDB and GenBank. AB - Database interconnection requires the development of links between related objects from different databases. We built a database of links, called Virgil, to manage and distribute rich (documented) links between GDB genes and GenBank human sequences. Virgil contains 18 667 unique links. In addition to a simple Web form for ad-hoc queries, we propose a generic Web interface and a prototype CORBA server for link distribution. Materials described in this paper are available from http://www.infobiogen.fr/services/virgil/home. html PMID- 9399808 TI - GDB: the Human Genome Database. AB - The Genome Database (GDB, http://www.gdb.org ) is a public repository of data on human genes, clones, STSs, polymorphisms and maps. GDB entries are highly cross linked to each other, to literature citations and to entries in other databases, including the sequence databases, OMIM, and the Mouse Genome Database. Mapping data from large genome centers and smaller mapping efforts are added to GDB on an ongoing basis. The database can be searched by a variety of methods, ranging from keyword searches to complex queries. Major functionality extensions in the last year include the ongoing computation of integrated human genome maps, called Comprehensive Maps, and the use of those maps to support positional queries and graphic displays. The capabilities of the GDB map viewer (Mapview) have been extended to include map printing and the graphical display of ad hoc query results. The HUGO Nomenclature Committee continues to curate the proposed and official gene symbols and related data in collaboration with GDB. As genome research shifts its emphasis from mapping to sequencing and functional analysis, the scope of the GDB schema is being extended. We are in the process of adding representations of gene function and expression, and improving our representation of human polymorphism and mutation. PMID- 9399810 TI - The Radiation Hybrid Database. AB - Since July 1995, the European Bioinformatics Institute (EBI) has maintained RHdb (http://www.ebi.ac.uk/RHdb/RHdb.html ), a public database for radiation hybrid data. Radiation hybrid mapping is an important technique for determining high resolution maps. Recently, CORBA access has been added to Rhdb. The EBI is an Outstation of the European Molecular Biology Laboratory (EMBL). PMID- 9399811 TI - HuGeMap: a distributed and integrated Human Genome Map database. AB - The HuGeMap database stores the major genetic and physical maps of the human genome. It is also interconnected with the gene radiation hybrid mapping database RHdb. HuGeMap is accessible through a Web server for interactive browsing at URL http://www.infobiogen. fr/services/Hugemap , as well as through a CORBA server for effective programming. HuGeMap is intended as an attempt to build open, interconnected databases, that is databases that distribute their objects worldwide in compliance with a recognized standard of distribution. Maps can be displayed and compared with a java applet (http://babbage.infobiogen.fr:15000/Mappet/Show. html ) that queries the HuGeMap ORB server as well as the RHdb ORB server at the EBI. PMID- 9399812 TI - IXDB, an X chromosome integrated database. AB - The integrated X chromosome database (IXDB) is a repository for physical mapping data of the human X chromosome. Its current content is the result of a strict integration of data stemming from many different sources. The main features of IXDB include a flexible and extendible schema, a comfortable and fully cross referenced WWW interface (http://ixdb.mpimg-berlin-dahlem.mpg.de ) and a graphical map viewer implemented in JAVA. The database stores objects used in physical mapping as well as the maps resulting from this work, but a strong emphasis is placed on recording experiments that connect objects together. This should greatly contribute to fulfilling one of the major goals of the database: to support the construction of an integrated physical, genetic, transcript and sequence map of the human X chromosome. PMID- 9399813 TI - MITOMAP: a human mitochondrial genome database--1998 update. AB - We have continued to develop MITOMAP (http://www.gen.emory. edu/MITOMAP ), a comprehensive database for the human mitochondrial DNA (mtDNA). MITOMAP uses the mtDNA sequence as the unifying element for bringing together information on mitochondrial genome structure and function, pathogenic mutations and their clinical characteristics, population associated variation, and gene-gene interactions. Over the past year we have increased the degree of interlinking of MITOMAP information available on the web page, by using our generalized information management system, GENOME. As increasingly larger regions of the human genome are sequenced and characterized, the need for integrating such information is growing. Consequently, MITOMAP and GENOME provide a valuable reference for the mitochondrial biologist, in addition to being a model for the development of comprehensive, information storage and retrieval systems for other components of the human genome. This paper documents the changes to MITOMAP which have been implemented over the past year. PMID- 9399814 TI - The mitBASE human dataset structure. AB - MitBASE is a comprehensive and integrated mitochondrial genome database funded within the EU BIOTECH PROGRAM. It is a project for the development and implementation of an integrated and comprehensive database of mitochondrial data which will collect all available information from different organisms and from intraspecies variants and mutants. The present paper describes the structure of the Human dataset in mitBASE where human molecular data are distinguished from clinical and pathological data. MitBASE home page address is: http://www.ebi.ac.uk/htbin/Mitbase/mitb ase.pl PMID- 9399815 TI - Update of MmtDB: a Metazoa mitochondrial DNA variants database. AB - The present paper describes the improvements in MmtDB, a specialised database designed to collect Metazoa mitochondrial DNA variants. Priority in the data collection has been given to Metazoa for which a large amount of variants is available, e.g., for humans. Starting from the sequences available in the Nucleotide Sequence Databases, the redundant sequences have been removed and new sequences from other sources have been added. Value-added information is associated to each variant sequence, e.g., analysed region, experimental method, tissue and cell lines, population data, sex, age, family code and information about the variation events (nucleotide position, involved gene, restriction site gain or loss). Cross-references are introduced to the EMBL Data Library, as well as an internal cross-referencing among MmtDB entries according to tissual, heteroplasmic, familiar and aplotypical correlation. Furthermore MmtDB has a new section, AMmtDB: Aligned Metazoan mitochondrial biosequences. MmtDB can be accessed through the World Wide Web at URL http://WWW.ba.cnr.it/[symbol: see text]areamt08/MmtDBWWW.htm PMID- 9399817 TI - The Mouse Genome Database (MGD): a community resource. Status and enhancements. The Mouse Genome Informatics Group. AB - The Mouse Genome Database (MGD) is a comprehensive community database that integrates genetic, genomic and phenotypic information about the laboratory mouse. MGD provides detailed information about genes and genetic markers, elemental data from mapping experiments, descriptions of molecular segments including ESTs, probes, and cDNA clones, homology information between mouse and many other mammalian genomes, and phenotypic descriptions of gene mutations, gene function and mouse strains. All data are supported by citations. Interactive graphical displays of cytogenetic, genetic and physical maps are available. User support is provided through dedicated staff, bulletin boards, and user documentation. MGD can be accessed at http://www.informatics.jax.org PMID- 9399816 TI - Compilation of human mtDNA control region sequences. AB - This paper describes the organisation of a database for human mitochondrial control-region sequences. The data are divided into three ASCII files that contain aligned sequences from the hypervariable region I (HVRI), from the hypervariable region II (HVRII), and the available information about the individuals, from whom the sequences stem. The current collection comprises 4079 HVRI and 969 HVRII sequences. From 728 individuals sequences of both HVRI and HVRII are available. For easy access, the collection is made available to the scientific community via World Wide Web at URL http://www.zi.biologie.uni muenchen.de/[symbol: see text]meyers/mtdna.html PMID- 9399818 TI - The Organelle Genome Database Project (GOBASE). AB - The taxonomically broad organelle genome database (GOBASE) organizes and integrates diverse data related to organelles (mitochondria and chloroplasts). The current version of GOBASE focuses on the mitochondrial subset of data and contains molecular sequences, RNA secondary structures and genetic maps, as well as taxonomic information for all eukaryotic species represented. The database has been designed so that complex biological queries, especially ones posed in a comparative genomics context, are supported. GOBASE has been implemented as a relational database with a web-based user interface (http://megasun.bch.umontreal.ca/gobase/gobas e.html ). Custom software tools have been written in house to assist in the population of the database, data validation, nomenclature standardization and front-end design. The database is fully operational and publicly accessible via the World Wide Web, allowing interactive browsing, sophisticated searching and easy downloading of data. PMID- 9399819 TI - Molecular Probe Data Base (MPDB). AB - In this paper, the current status of the Molecular Probe Data Base (http://www.biotech.ist.unige.it/interlab/ mpdb.html ) is briefly presented together with a short analysis of its activity during 1997. This has been performed by statistically evaluating the 'logs' of the Internet servers that are used for its distribution with reference to the geographical origin of the requests, the words that were utilized to carry out of the searches and the oligonucleotides that were retrieved. Planned enhancements of this database are also described. They include a revision of its data structure and, even more relevant, of its data management procedures. PMID- 9399820 TI - Compilation of tRNA sequences and sequences of tRNA genes. AB - Sequences of 3279 sequences of tRNA genes and tRNAs published up to December 1996 are included in the compilation. Alignment of the sequences, which is most compatible with the tRNA phylogeny and known three-dimensional structures of tRNA, is used. Sequences and references are available under http://www.uni bayreuth. de/departments/biochemie/trna/ PMID- 9399821 TI - A FastA based compilation of higher plant mitochondrial tRNA genes. AB - A new version of the compilation of higher plant mitochondrial tRNA genes (http://www.ebi.ac.uk/service ) has been obtained by means of the FastA program for similarity searching in nucleotide sequence Databases. This approach improves the previous collection, which was based on literature data analysis. The current compilation contains 158 sequences with an increase of 43 units. In this paper, some interesting features of the new entries are briefly presented. PMID- 9399822 TI - 5S rRNA Data Bank. AB - In this paper we present the updated version of the compilation of 5S rRNA and 5S rDNA nucleotide sequences. It contains 1622 primary structures of 5S rRNAs and 5S rRNA genes from 888 species. These include 58 archaeal, 427 eubacterial, 34 plastid, nine mitochondrial and 1094 eukaryotic DNA or RNA nucleotide sequences. The sequence entries are divided according to the taxonomic position of the organisms. All individual sequences deposited in the 5S rRNA Database can be retrieved using the WWW-based, taxonomic browser at http://rose.man.poznan.pl/5SData/5SRNA.html++ + or http://www.chemie. fu berlin.de/fb_chemie/agerdmann/5S_rRNA.html . The files with complete sets of data as well as sequence alignments are available via anonymous ftp. PMID- 9399823 TI - Small RNA database. AB - The small RNA database is a compilation of all the small size RNA sequences available to date, including nuclear, nucleolar, cytoplasmic and mitochondria small RNAs from eukaryotic organisms and small RNAs from prokaryotic cells as well as viruses. Currently, approximately 600 small RNA sequences are in our database. It also gives the sources of individual RNAs and their GenBank accession numbers. The small RNA database can be accessed through the WWW (World Wide Web). Our WWW URL address is: http://mbcr.bcm.tmc. edu/smallRNA/smallrna.html . The new small RNA sequences published since our last compilation are listed in this paper (Table 1). PMID- 9399825 TI - The tmRNA database (tmRDB). AB - This first release of the tmRNA database (tmRDB) contains 19 tmRNA sequences, a tmRNA sequence alignment with emphasis of base pairs that are supported by comparative sequence analysis, and a tabulation of tmRNA-encoded tag peptides. The tmRNADB also offers an RNA secondary structure diagram of the Escherichia coli tmRNA, as well as PDB-formatted coordinates for three-dimensional modeling. The data are available on the World Wide Web at http://www.uthct. edu/tmRDB/tmRDB.html PMID- 9399824 TI - The tmRNA Website. AB - tmRNA (also known as 10Sa RNA) is so-named for its dual tRNA-like and mRNA-like nature. It is employed in a remarkable trans -translation process to add a C terminal peptide tag to the incomplete protein product of a broken mRNA; the tag targets the abnormal protein for proteolysis. tmRNA sequences have been identified in genomes of diverse bacterial phyla, including the most deeply branching. They have also been identified in plastids of the 'red' lineage. The tmRNA Website (http://www.wi.mit. edu/bartel/tmRNA/home ) contains a database currently including sequences from 37 species, with provisional alignments, as well as the tentatively predicted proteolysis tag sequences. A brief review and guide to the literature is also provided. PMID- 9399826 TI - The guide RNA database. AB - Guide RNAs (gRNAs) are small, metabolically stable RNA molecules which perform a pivotal, template-like function during the RNA editing process in kinetoplastid protozoa. The gRNA database currently contains 250 guide RNA sequences as well as secondary and tertiary structure models and other relevant information. The database is made available as a hypertext document accessible via the World Wide Web (WWW) at the URL: http://www.biochem.mpg.de/ goeringe/ PMID- 9399827 TI - U-insertion/deletion Edited Sequence Database. AB - Uridine insertion/deletion RNA editing is a post-transcriptional RNA modification occurring in the mitochondria of kinetoplastid protozoa. The U-insertion/deletion Edited Sequence Database is a compilation of mitochondrial genes and edited mRNAs from five kinetoplastid species. It contains separate files with the DNA, mRNA (both unedited and edited) and predicted protein sequences, as well as alignments of the Leishmania tarentolae and Trypanosoma brucei protein sequences from edited and unedited genes. The sequence files are in GCG format. A 'map' sequence file showing the location of U-deletions, U-insertions and the translated amino acid sequences is also provided for each gene. Genomic maps for each species are also provided with clickable genes, including maxicircle-encoded gRNAs. Sets of aligned nuclear rRNA sequences from kinetoplastid protozoa are also provided, which were used for phylogenetic reconstructions in an analysis of the origin of RNA editing. The database is available through the World Wide Web as an HTML document at the URLhttp://www.lifesci.ucla.edu/RNA/trypanosome/ database.html PMID- 9399828 TI - The Signal Recognition Particle Database (SRPDB). AB - This release of the SRPDB (signal recognition particle database, http://pegasus.uthct.edu/SRPDB/SRPDB . html ) adds four SRP RNA sequences (a total of 99 SRP RNA sequences), 23 SRP protein sequences (a total of 63 protein sequences from SRP9, SRP14, SRP19, SRP21, SRP54, SRP68 or SRP72), and, for the first time, sequences of the alpha subunit of the eukaryotic SRP receptor and its homologous bacterial proteins (a total of 21 sequences). Sequences are offered phylogenetically ordered, annotated with links to the primary databases, and in aligned form. Also downloadable are sample SRP RNA secondary structure diagrams, three-dimensional models of representative SRP RNAs, and search motifs. PMID- 9399829 TI - Database on the structure of small ribosomal subunit RNA. AB - About 8600 complete or nearly complete sequences are now available from the Antwerp database on small ribosomal subunit RNA. All these sequences are aligned with one another on the basis of the adopted secondary structure model, which is corroborated by the observation of compensating substitutions in the alignment. Literature references, accession numbers and detailed taxonomic information are also compiled. The database can be consulted via the World Wide Web at URL http://rrna.uia.ac.be/ssu/ PMID- 9399830 TI - Database on the structure of large ribosomal subunit RNA. AB - The rRNA WWW Server at URL http://rrna.uia.ac.be/ now provides a database of 496 large subunit ribosomal RNA sequences. All these sequences are aligned, incorporate secondary structure information, and can be obtained in a number of formats. Other information about the sequences, such as literature references, accession numbers and taxonomic information is also available and searchable. If necessary, the data on the server can also be obtained by anonymous ftp. PMID- 9399831 TI - The Database of Ribosomal Cross links (DRC). AB - The Database of Ribosomal Cross-links (DRC) provides a complete collection of all the published data produced by cross-linking studies on the Escherichia coli ribosome, as well as on its components and functional ligands. The DRC currently includes data on 986 cross-links from >100 research papers, yielded by >40 different reagents. For each cross-link, information is given concerning its location in the ribosome, the chemical or photochemical reagent applied, a brief description of the method(s) used to locate the cross-link, and the literature reference. The DRC is freely available via the World Wide Web at: http://Ribosome.Genebee.MSU.SU/DRC/ or at http://WWW:MPIMG-Berlin Dahlem.MPG.DE/[symbol: see text]baranov/DRC/ PMID- 9399832 TI - The viroid and viroid-like RNA database. AB - The viroid and viroid-like RNA database is a compilation of all natural sequences published in journals or available from the GenBank and EMBL nucleotide sequence libraries. Several information regarding these RNA species such as the position of their self-catalytic domains and the open reading frame of the human hepatitits delta virus are provided. The database also includes a determination of the likely ancestral sequence of most species and a prediction of the most stable secondary structures of these sequences. This online database is available on the World Wide Web (http://www.callisto.si.usherb.ca/[symbol: see text]jpperra ). It should provide an excellent reference point for further phylogenetic and structure-function studies of these RNA species. PMID- 9399833 TI - UTRdb: a specialized database of 5'- and 3'-untranslated regions of eukaryotic mRNAs. AB - The important role the untranslated regions of eukaryotic mRNAs may play in gene regulation and expression is now widely acknowledged. For this reason we developed UTRdb, a specialized database of 5'- and 3'-untranslated sequences of eukaryotic mRNAs cleaned from redundancy. UTRdb entries are enriched with specialized information not present in the primary databases, including the presence of functional patterns already demonstrated by experimental analysis to have some functional role. A collection of such patterns is being collected in UTRsite database (http://bio-www.ba.cnr.it:8000/srs5/) which can also be used with appropriate computational tools to detect known functional patterns contained in mRNA untranslated regions. PMID- 9399834 TI - The RNA modification database--1998. AB - The RNA modification database provides a comprehensive listing of posttranscriptionally modified nucleosides from RNA, and is maintained as an updated version of the initial printed report [Limbach,P.A., Crain,P.F. and McCloskey,J.A. (1994) Nucleic Acids Res. , 22, 2183-2196]. Information provided for each nucleoside includes: the type of RNA in which it occurs and phylogenetic distribution; common chemical name and symbol; Chemical Abstracts registry number and index name; chemical structure; initial literature citations for structural characterization or occurrence, and for chemical synthesis. The data are available through the World Wide Web at: http://www medlib.med.utah/RNAmods/RNAmods .html PMID- 9399835 TI - Databases and software for the analysis of mutations in the human p53 gene, human hprt gene and both the lacI and lacZ gene in transgenic rodents. AB - We have created databases and software applications for the analysis of DNA mutations at the human p53 gene, the human hprt gene and both the rodent transgenic lacI and lacZ loci. The databases themselves are stand-alone dBASE files and the software for analysis of the databases runs on IBM-compatible computers with Microsoft Windows. Each database has a separate software analysis program. The software created for these databases permit the filtering, ordering, report generation and display of information in the database. In addition, a significant number of routines have been developed for the analysis of single base substitutions. One method of obtaining the databases and software is via the World Wide Web. Open the following home page with a Web Browser: http://sunsite.unc.edu/dnam/mainpage. html . Alternatively, the databases and programs are available via public FTP from: anonymous@sunsite.unc.edu. There is no password required to enter the system. The databases and software are found beneath the subdirectory: pub/academic/biology/dna-mutations. Two other programs are available at the site, a program for comparison of mutational spectra and a program for entry of mutational data into a relational database. PMID- 9399836 TI - p53 gene mutation: software and database. AB - A large number of different mutations in the p53 tumor suppressor gene have been identified in all types of cancer. As of October, 1997, this database (http:// perso.curie.fr/tsoussi ) contained >7500 mutations. Such a substantial increase since our previous reports should enable epidemiological analyses which were not previously possible. In order to analyse these new data, the UMD software has been improved. A new Web version of the UMD software enables online analysis of the database. The present report describes various improvements since the last release of the database. PMID- 9399837 TI - IARC Database of p53 gene mutations in human tumors and cell lines: updated compilation, revised formats and new visualisation tools. AB - Since 1989, about 570 different p53 mutations have been identified in more than 8000 human cancers. A database of these mutations was initiated by M. Hollstein and C. C. Harris in 1990. This database originally consisted of a list of somatic point mutations in the p 53 gene of human tumors and cell lines, compiled from the published literature and made available in a standard electronic form. The database is maintained at the International Agency for Research on Cancer (IARC) and updated versions are released twice a year (January and July). The current version (July 1997) contains records on 6800 published mutations and will surpass the 8000 mark in the January 1998 release. The database now contains information on somatic and germline mutations in a new format to facilitate data retrieval. In addition, new tools are constructed to improve data analysis, such as a Mutation Viewer Java applet developed at the European Bioinformatics Institute (EBI) to visualise the location and impact of mutations on p53 protein structure. The database is available in different electronic formats at IARC (http://www.iarc. fr/p53/homepage.htm ) or from the EBI server (http://www.ebi.ac.uk ). The IARC p53 website also provides reports on database analysis and links with other p53 sites as well as with related databases. In this report, we describe the criteria for inclusion of data, the revised format and the new visualisation tools. We also briefly discuss the relevance of p 53 mutations to clinical and biological questions. PMID- 9399838 TI - A database of germline p53 mutations in cancer-prone families. AB - We created a comprehensive database covering all published cases of germline p53 mutations. The current version lists 580 tumours in 448 individuals belonging to 122 independent pedigrees. The database describes each p53 mutation (type of the mutation, exon and codon affected by the mutation, nucleotide and amino acid change), each family (family history of cancer, diagnosis of Li-Fraumeni syndrome), each affected individual (sex, generation, p53 status, from which parent the mutation was inherited) and each tumour (type, age of onset, p53 status-loss of heterozygosity, immunostaining). Each entry contains the original reference(s). The database is freely available and can be obtained from http://www.lf2.cuni.cz PMID- 9399839 TI - The factor VIII Structure and Mutation Resource Site: HAMSTeRS version 4. AB - Since 1996 the HAMSTeRS (Haemophilia A Mutation, Search, Test and Resource Site) WWW site has provided an online resource for access to data on the molecular pathology of haemophilia A, replacing previous text editions of the Haemophilia A Database published in Nucleic Acids Research . This report describes the continued development of the site (version 4), and in particular the expansion of factor VIII (FVIII) structure-related features. Access to the mutation database itself, both for searching the listings and for submission of new mutations, is via custom-designed forms: more powerful Boolean searches of the point mutations in the database are also available. During 1997 a total of 22 novel missense mutations were reported, increasing the total number of unique variants now described to 252 (238 in exonic sequences and 14 at intronic splice junctions). Currently, a total of 586 individual reports with associated phenotypic data are available for searching by any category including phenotype. The FVIII structure section now includes a download of a FVIII A domain homology model in Protein Data Bank format and a multiple alignment of the FVIII amino-acid sequencies from four species (human, murine, porcine and canine) in addition to the virtual reality simulations, secondary structural data and FVIII animation already available. Finally, to aid navigation across this site, a clickable roadmap of the main features provides easy access to the page desired. Our intention is that continued development and updating of the site shall provide workers in the fields of molecular and structural biology with a one-stop resource site to facilitate FVIII research and education. The HAMSTeRS URL is http://europium.mrc.rpms.ac.uk PMID- 9399840 TI - PAH Mutation Analysis Consortium Database: 1997. Prototype for relational locus specific mutation databases. AB - PAHdb (http://www.mcgill.ca/pahdb ) is a curated relational database (Fig. 1) of nucleotide variation in the human PAH cDNA (GenBank U49897). Among 328 different mutations by state (Fig. 2) the majority are rare mutations causing hyperphenylalaninemia (HPA) (OMIM 261600), the remainder are polymorphic variants without apparent effect on phenotype. PAHdb modules contain mutations, polymorphic haplotypes, genotype-phenotype correlations, expression analysis, sources of information and the reference sequence; the database also contains pages of clinical information and data on three ENU mouse orthologues of human HPA. Only six different mutations account for 60% of human HPA chromosomes worldwide, mutations stratify by population and geographic region, and the Oriental and Caucasian mutation sets are different (Fig. 3). PAHdb provides curated electronic publication and one third of its incoming reports are direct submissions. Each different mutation receives a systematic (nucleotide) name and a unique identifier (UID). Data are accessed both by a Newsletter and a search engine on the website; integrity of the database is ensured by keeping the curated template offline. There have been >6500 online interrogations of the website. PMID- 9399841 TI - aCHEdb: the database system for ESTHER, the alpha/beta fold family of proteins and the Cholinesterase gene server. AB - Acetylcholinesterase belongs to a family of proteins, the alpha/beta hydrolase fold family, whose constituents evolutionarily diverged from a common ancestor and share a similar structure of a central beta sheet surrounded by alpha helices. These proteins fulfil a wide range of physiological functions (hydrolases, adhesion molecules, hormone precursors) [Krejci,E., Duval,N., Chatonnet,A., Vincens,P. and Massoulie,J. (1991) Proc. Natl. Acad. Sci. USA , 88, 6647-6651]. ESTHER (for esterases, alpha/beta hydrolase enzymes and relatives) is a database aimed at collecting in one information system, sequence data together with biological annotations and experimental biochemical results related to the structure-function analysis of the enzymes of the family. The major upgrade of the database comes from the use of a new database management system: aCHEdb which uses the ACeDB program designed by Richard Durbin and Jean Thierry-Mieg. It can be found at http://www.ensam.inra.fr/cholinesterase PMID- 9399842 TI - Marfan Database (third edition): new mutations and new routines for the software. AB - The Marfan database is a software that contains routines for the analysis of mutations identified in the FBN1 gene that encodes fibrillin-1. Mutations in this gene are associated not only with Marfan syndrome but also with a spectrum of overlapping disorders. The third version of the Marfan database contains 137 entries. The software has been modified to accommodate four new routines and is now accessible on the World Wide Web at http://www.umd.necker.fr PMID- 9399843 TI - The Androgen Receptor Gene Mutations Database. AB - The current version of the androgen receptor (AR) gene mutations database is described. The total number of reported mutations has risen from 272 to 309 in the past year. We have expanded the database: (i) by giving each entry an accession number; (ii) by adding information on the length of polymorphic polyglutamine (polyGln) and polyglycine (polyGly) tracts in exon 1; (iii) by adding information on large gene deletions; (iv) by providing a direct link with a completely searchable database (courtesy EMBL-European Bioinformatics Institute). The addition of the exon 1 polymorphisms is discussed in light of their possible relevance as markers for predisposition to prostate or breast cancer. The database is also available on the internet (http://www.mcgill. ca/androgendb/ ), from EMBL-European Bioinformatics Institute (ftp. ebi.ac.uk/pub/databases/androgen ), or as a Macintosh FilemakerPro or Word file (MC33@musica.mcgill.ca). PMID- 9399844 TI - BTKbase, mutation database for X-linked agammaglobulinemia (XLA). AB - X-linked agammaglobulinemia (XLA) is an immunodeficiency caused by mutations in the gene coding for Bruton's agammaglobulinemia tyrosine kinase (BTK). A database (BTKbase) of BTK mutations has been compiled and the recent update lists 463 mutation entries from 406 unrelated families showing 303 unique molecular events. In addition to mutations, the database also lists variants or polymorphisms. Each patient is given a unique patient identity number (PIN). Information is included regarding the phenotype including symptoms. Mutations in all the five domains of BTK have been noticed to cause the disease, the most common event being missense mutations. The mutations appear almost uniformly throughout the molecule and frequently affect CpG sites that code for arginine residues. The putative structural implications of all the missense mutations are given in the database. The improved version of the registry having a number of new features is available at http://www. helsinki.fi/science/signal/btkbase.html PMID- 9399845 TI - LDLR Database (second edition): new additions to the database and the software, and results of the first molecular analysis. AB - Mutations in the LDL receptor gene (LDLR) cause familial hypercholesterolemia (FH), a common autosomal dominant disorder. The LDLR database is a computerized tool that has been developed to provide tools to analyse the numerous mutations that have been identified in the LDLR gene. The second version of the LDLR database contains 140 new entries and the software has been modified to accommodate four new routines. The analysis of the updated data (350 mutations) gives the following informations: (i) 63% of the mutations are missense, and only 20% occur in CpG dinucleotides; (ii) although the mutations are widely distributed throughout the gene, there is an excess of mutations in exons 4 and 9, and a deficit in exons 13 and 15; (iii) the analysis of the distribution of mutations located within the ligand-binding domain shows that 74% of the mutations in this domain affect a conserved amino-acid, and that they are mostly confined in the C-terminal region of the repeats. Conversely, the same analysis in the EGF-like domain shows that 64% of the mutations in this domain affect a non-conserved amino-acid, and, that they are mostly confined in the N-terminal half of the repeats. The database is now accessible on the World Wide Web at http://www.umd.necker.fr PMID- 9399846 TI - The Human Collagen Mutation Database 1998. AB - The collagens are a large and diverse family of proteins which are found in the extracellular matrix. In common with one another, the 19 known collagen types have triple-helical domains of variable length but they differ with respect to their overall size and the nature and location of their globular domains. Collagen mutations lead to heritable defects of connective tissues and mutation data for collagen types I and III are presented here. The mutation data are accessible on the world wide web at http://www.le.ac.uk/genetics/collagen/ PMID- 9399847 TI - Software and database for the analysis of mutations in the VHL gene. AB - VHL is a tumor suppressor gene localized on chromosome 3p25-26. Mutations of the VHL gene were described at first in the heritable von Hippel-Lindau disease and in the sporadic Renal Cell Carcinoma (RCC). More recently, VHL has also been shown to harbor mutations in mesothelioma and small cell lung carcinoma. To date more than 500 mutations have been identified. These mutations are mainly private with only one hot spot at codon 167 associated with pheochromocytoma. The germline mutations are essentially missense while somatic mutations include deletions, insertions and nonsense. To standardize the collection of these informations, facilitate the mutational analysis of the VHL gene and promote the genotype-phenotype analysis, a software package along with a computerized database have been created. The current database and the analysis software are accessible via the internet and world wide web interface at the URL:http://www.umd.necker.fr PMID- 9399848 TI - The Human PAX6 Mutation Database. AB - The Human PAX6 Mutation Database contains details of 94 mutations of the PAX6 gene. A Microsoft Access program is used by the Curator to store, update and search the database entries. Mutations can be entered directly by the Curator, or imported from submissions made via the World Wide Web. The PAX6 Mutation Database web page at URL http://www.hgu.mrc.ac.uk/Softdata/PAX6/ provides information about PAX6, as well as a fill-in form through which new mutations can be submitted to the Curator. A search facility allows remote users to query the database. A plain text format file of the data can be downloaded via the World Wide Web. The Curation program contains prior knowledge of the genetic code and of the PAX6 gene including cDNA sequence, location of intron/exon boundaries, and protein domains, so that the minimum of information need be provided by the submitter or Curator. PMID- 9399849 TI - Haemophilia B: database of point mutations and short additions and deletions- eighth edition. AB - The eighth edition of the haemophilia B database (http://www.umds.ac. uk/molgen/haemBdatabase.htm ) lists in an easily accessible form all known factor IX mutations due to small changes (base substitutions and short additions and/or deletions of <30 bp) identified in haemophilia B patients. The 1713 patient entries are ordered by the nucleotide number of their mutation. Where known, details are given on: factor IX activity, factor IX antigen in circulation, presence of inhibitor and origin of mutation. References to published mutations are given and the laboratories generating the data are indicated. PMID- 9399850 TI - APC gene: database of germline and somatic mutations in human tumors and cell lines. AB - A database (http://perso.curie.fr/tsoussi ) is described, in which over 1000 mutations in the human APC gene of tumors (colon cancer predominantly) are compiled from the literature. It includes both molecular information about the mutations and clinical data about the patients. Software has been designed to analyse all this information in the database. PMID- 9399851 TI - Software and database for the analysis of mutations in the human WT1 gene. AB - The WT1 gene, located at 11p13, encodes a zinc finger transcription factor involved in renal and gonadal development and in Wilms' tumor. Constitutional mutations of this gene have been described in most patients with Denys Drash syndrome (mesangial sclerosis associated with male pseudohermaphrodism and/or Wilms' tumor), but also in patients with genitourinary abnormalities and Wilms' tumor (WT) or presenting with only unilateral or bilateral WT. Moreover, approximately 10% of Wilms' tumors carry WT1 mutations at the somatic level. To facilitate the genotype-phenotype correlation analyses, we have created a software package along with a computerized database of germline (70 entries) and somatic (28 entries) mutations reported in the literature. PMID- 9399852 TI - GPCRDB: an information system for G protein-coupled receptors. AB - The GPCRDB is a G protein-coupled receptor (GPCR) database system aimed at the collection and dissemination of GPCR related data. It holds sequences, mutant data and ligand binding constants as primary (experimental) data. Computationally derived data such as multiple sequence alignments, three dimensional models, phylogenetic trees and two dimensional visualization tools are added to enhance the database's usefulness. The GPCRDB is an EU sponsored project aimed at building a generic molecular class specific database capable of dealing with highly heterogeneous data. GPCRs were chosen as test molecules because of their enormous importance for medical sciences and due to the availability of so much highly heterogeneous data. The GPCRDB is available via the WWW at http://www.gpcr.org/7tm PMID- 9399853 TI - Expanded versions of the 16S and 23S ribosomal RNA mutation databases (16SMDBexp and 23SMDBexp) AB - Expanded versions of the Ribosomal RNA Mutation Databases provide lists of mutated positions in 16S and 16S-like ribosomal RNA (16SMDBexp) and 23S and 23S like ribosomal RNA (23SMDBexp) and the identity of each alteration. Alterations from organisms other than Escherichia coli are reported at positions according to the E.coli numbering system. Information provided for each mutation includes: (i) a brief description of the phenotype(s) associated with each mutation, (ii) whether a mutant phenotype has been detected by in vivo or in vitro methods, and (iii) relevant literature citations. The databases are available via ftp and on the World Wide Web at the following URL: http: //www.fandm.edu/Departments/Biology/Databases/RNA.h tml PMID- 9399854 TI - The human gene mutation database. AB - The Human Gene Mutation Database (HGMD) represents a comprehensive core collection of data on published germline mutations in nuclear genes underlying human inherited disease. By September 1997, the database contained nearly 12 000 different lesions in a total of 636 different genes, with new entries currently accumulating at a rate of over 2000 per annum. Although originally established for the scientific study of mutational mechanisms in human genes, HGMD has acquired a much broader utility to researchers, physicians and genetic counsellors so that it was made publicly available at http://uwcm.ac.uk/uwcm/mg/hgmd0.html in April 1996. Mutation data in HGMD are accessible on the basis of every gene being allocated one web page per mutation type, if data of that type are present. Meaningful integration with phenotypic, structural and mapping information has been accomplished through bi-directional links between HGMD and both the Genome Database (GDB) and Online Mendelian Inheritance in Man (OMIM), Baltimore, USA. Hypertext links have also been established to Medline abstracts through Entrez , and to a collection of 458 reference cDNA sequences also used for data checking. Being both comprehensive and fully integrated into the existing bioinformatics structures relevant to human genetics, HGMD has established itself as the central core database of inherited human gene mutations. PMID- 9399855 TI - EpoDB: a database of genes expressed during vertebrate erythropoiesis. AB - EpoDB is a database designed for the study of gene regulation during differentiation and development of vertebrate red blood cells. In building EpoDB, we have taken the in advance approach to the data integration problem: we have extracted data relevant to red blood cells from GenBank, SWISS-PROT, TRRD (transcriptional regulation data) and GERD (expression levels data) to create a single integrated, highly curated view. Tools have been developed to automate data extraction from online resources, cleanse data of errors, enter information manually from the primary literature, generate a uniform, canonical representation of information and maintain data currency. The database is organized around biological features, e.g., genes, rather than sequences, which are supported by a controlled and consistent vocabulary for gene names and gene family names. Beyond the standard database queries, the functionality of EpoDB includes the ability to extract features and subsequences, display sequences and features graphically using bioWidget viewers and integrated analysis tools. EpoDB may be accessed at: http://cbil.humgen.upenn.edu/epodb/ PMID- 9399856 TI - A mutation spectra database for bacterial and mammalian genes: 1998. AB - This database consists of over 24 000 mutations in 18 viral, bacterial, yeast or mammalian genes. The data are grouped as sets of DNA base sequence changes or spectra caused by a particular mutagen under defined conditions. The spectra are available on the World Wide Web at http://info.med.yale.edu/mutbase/ in two formats; in text format that can be browsed on-line or downloaded for use with a text editor and in dBASEIII format for use, after downloading, by relational database programs or by spreadsheets. Researchers are encouraged to submit DNA sequence changes to a suitable mutation database such as ours. A data entry program, MUTSIN, can be retrieved from this site. MUTSIN diagrams each mutation on the computer screen and alerts the user to any discrepancies. PMID- 9399857 TI - Database of mutations within the adenovirus 5 E1A oncogene. AB - The Ad5 E1A database is a listing of mutations affecting the early region 1A (E1A) proteins of human adenovirus type 5. The database contains the name of the mutation, the nucleic acid sequence changes, the resulting alterations in amino acid sequence and reference. Additional notes and references are provided on the effect of each mutation on E1A function. The database is contained within the Adenovirus 5 E1A page on the World Wide Web at: http://www.geocities.com/CapeCanaveral/Hangar /2541/ PMID- 9399858 TI - SV40 large tumor antigen (T antigen): database of mutants. AB - The SV40 T antigen database (http://www.pitt.edu/pipaslab/) lists viruses and plasmids expressing mutant forms of large T antigen. Each entry contains information regarding the mutant designation, mutant type, virus strain, nucleotide change, amino acid change and pertinent references. The database is now available as an internet searchable index. PMID- 9399859 TI - IMGT, the International ImMunoGeneTics database. AB - IMGT, the international ImMunoGeneTics database, is an integrated database specialising in Immunoglobulins (Ig), T cell Receptors (TcR) and Major Histocompatibility Complex (MHC) of all vertebrate species, created by Marie Paule Lefranc, CNRS, Montpellier II University, Montpellier, France (lefranc@ligm.crbm.cnrs-mop.fr). IMGT includes three databases: LIGM-DB (for Ig and TcR), MHC/HLA-DB and PRIMER-DB (the last two in development). IMGT comprises expertly annotated sequences and alignment tables. LIGM-DB contains more than 23 000 Immunoglobulin and T cell Receptor sequences from 78 species. MHC/HLA-DB contains Class I and Class II Human Leucocyte Antigen alignment tables. An IMGT tool, DNAPLOT, developed for Ig, TcR and MHC sequence alignments, is also available. IMGT works in close collaboration with the EMBL database. IMGT goals are to establish a common data access to all immunogenetics data, including nucleotide and protein sequences, oligonucleotide primers, gene maps and other genetic data of Ig, TcR and MHC molecules, and to provide a graphical user friendly data access. IMGT has important implications in medical research (repertoire in autoimmune diseases, AIDS, leukemias, lymphomas), therapeutical approaches (antibody engineering), genome diversity and genome evolution studies. IMGT is freely available at http://imgt.cnusc.fr:8104 PMID- 9399860 TI - The PRINTS protein fingerprint database in its fifth year. AB - PRINTS is a database of protein family 'fingerprints' offering a diagnostic resource for newly-determined sequences. By contrast with PROSITE, which uses single consensus expressions to characterise particular families, PRINTS exploits groups of motifs to build characteristic signatures. These signatures offer improved diagnostic reliability by virtue of the mutual context provided by motif neighbours. To date, 800 fingerprints have been constructed and stored in PRINTS. The current version, 17.0, encodes approximately 4500 motifs, covering a range of globular and membrane proteins, modular polypeptides, and so on. The database is accessible via the UCL Bioinformatics World Wide Web (WWW) Server at http://www. biochem.ucl.ac.uk/bsm/dbbrowser/ . We have recently enhanced the usefulness of PRINTS by making available new, intuitive search software. This allows both individual query sequence and bulk data submission, permitting easy analysis of single sequences or complete genomes. Preliminary results indicate that use of the PRINTS system is able to assign additional functions not found by other methods, and hence offers a useful adjunct to current genome analysis protocols. PMID- 9399861 TI - Superior performance in protein homology detection with the Blocks Database servers. AB - The Blocks Database World Wide Web (http://www.blocks.fhcrc.org ) and Email (blocks@blocks.fhcrc.org) servers provide tools for the detection and analysis of protein homology based on alignment blocks representing conserved regions of proteins. During the past year, searching has been augmented by supplementation of the Blocks Database with blocks from the Prints Database, for a total of 4754 blocks from 1163 families. Blocks from both the Blocks and Prints Databases and blocks that are constructed from sequences submitted to Block Maker can be used for blocks-versus-blocks searching of these databases with LAMA, and for viewing logos and bootstrap trees. Sensitive searches of up-to-date protein sequence databanks are carried out via direct links to the MAST server using position specific scoring matrices and to the BLAST and PSI-BLAST servers using consensus embedded sequence queries. Utilizing the trypsin family to evaluate performance, we illustrate the superiority of blocks-based tools over expert pairwise searching or Hidden Markov Models. PMID- 9399862 TI - The HSSP database of protein structure-sequence alignments and family profiles. AB - HSSP (http: //www.sander.embl-ebi.ac.uk/hssp/) is a derived database merging structure (3-D) and sequence (1-D) information. For each protein of known 3D structure from the Protein Data Bank (PDB), we provide a multiple sequence alignment of putative homologues and a sequence profile characteristic of the protein family, centered on the known structure. The list of homologues is the result of an iterative database search in SWISS-PROT using a position-weighted dynamic programming method for sequence profile alignment (MaxHom). The database is updated frequently. The listed putative homologues are very likely to have the same 3D structure as the PDB protein to which they have been aligned. As a result, the database not only provides aligned sequence families, but also implies secondary and tertiary structures covering 33% of all sequences in SWISS PROT. PMID- 9399863 TI - Touring protein fold space with Dali/FSSP. AB - The FSSP database and its new supplement, the Dali Domain Dictionary, present a continuously updated classification of all known 3D protein structures. The classification is derived using an automatic structure alignment program (Dali) for the all-against-all comparison of structures in the Protein Data Bank. From the resulting enumeration of structural neighbours (which form a surprisingly continuous distribution in fold space) we derive a discrete fold classification in three steps: (i) sequence-related families are covered by a representative set of protein chains; (ii) protein chains are decomposed into structural domains based on the recurrence of structural motifs; (iii) folds are defined as tight clusters of domains in fold space. The fold classification, domain definitions and test sets for sequence-structure alignment (threading) are accessible on the web at www.embl-ebi.ac.uk/dali . The web interface provides a rich network of links between neighbours in fold space, between domains and proteins, and between structures and sequences leading, for example, to a database of explicit multiple alignments of protein families in the twilight zone of sequence similarity. The Dali/FSSP organization of protein structures provides a map of the currently known regions of the protein universe that is useful for the analysis of folding principles, for the evolutionary unification of protein families and for maximizing the information return from experimental structure determination. PMID- 9399864 TI - Pfam: multiple sequence alignments and HMM-profiles of protein domains. AB - Pfam contains multiple alignments and hidden Markov model based profiles (HMM profiles) of complete protein domains. The definition of domain boundaries, family members and alignment is done semi-automatically based on expert knowledge, sequence similarity, other protein family databases and the ability of HMM-profiles to correctly identify and align the members. Release 2.0 of Pfam contains 527 manually verified families which are available for browsing and on line searching via the World Wide Web in the UK at http://www.sanger.ac.uk/Pfam/ and in the US at http://genome.wustl. edu/Pfam/ Pfam 2.0 matches one or more domains in 50% of Swissprot-34 sequences, and 25% of a large sample of predicted proteins from the Caenorhabditis elegans genome. PMID- 9399865 TI - The ProDom database of protein domain families. AB - The ProDom database contains protein domain families generated from the SWISS PROT database by automated sequence comparisons. It can be searched on the World Wide Web (http://protein.toulouse.inra. fr/prodom.html ) or by E-mail (prodom@toulouse.inra.fr) to study domain arrangements within known families or new proteins. Strong emphasis has been put on the graphical user interface which allows for interactive analysis of protein homology relationships. Recent improvements to the server include: ProDom search by keyword; links to PROSITE and PDB entries; more sensitive ProDom similarity search with BLAST or WU-BLAST; alignments of query sequences with homologous ProDom domain families; and links to the SWISS-MODEL server (http: //www.expasy.ch/swissmod/SWISS-MODEL.html ) for homology based 3-D domain modelling where possible. PMID- 9399866 TI - An Integrated Sequence-Structure Database incorporating matching mRNA sequence, amino acid sequence and protein three-dimensional structure data. AB - We have constructed a non-homologous database, termed the Integrated Sequence Structure Database (ISSD) which comprises the coding sequences of genes, amino acid sequences of the corresponding proteins, their secondary structure and straight phi,psi angles assignments, and polypeptide backbone coordinates. Each protein entry in the database holds the alignment of nucleotide sequence, amino acid sequence and the PDB three-dimensional structure data. The nucleotide and amino acid sequences for each entry are selected on the basis of exact matches of the source organism and cell environment. The current version 1.0 of ISSD is available on the WWW at http://www.protein.bio.msu.su/issd/ and includes 107 non homologous mammalian proteins, of which 80 are human proteins. The database has been used by us for the analysis of synonymous codon usage patterns in mRNA sequences showing their correlation with the three-dimensional structure features in the encoded proteins. Possible ISSD applications include optimisation of protein expression, improvement of the protein structure prediction accuracy, and analysis of evolutionary aspects of the nucleotide sequence-protein structure relationship. PMID- 9399867 TI - Current status of the SWISS-2DPAGE database. AB - The SWISS-2DPAGE database (http: //www.expasy.ch/ch2d/ch2d-top.html ) consists of two-dimensional polyacrylamide gel electrophoresis images, as well as textual descriptions of the proteins that have been identified on them. The current release contains 15 reference maps from human biological samples, as well as from Saccharomyces cerevisiae , Escherichia coli and Dictyostelium discoideum origin. These reference maps have 2088 identified spots, corresponding to 410 separate protein entries in the database, in addition to virtual entries for each SWISS PROT sequence. PMID- 9399868 TI - Codon usage tabulated from the international DNA sequence databases. AB - CUTG (codon usage tabulated from GenBank) is a comprehensive database for codon usage. The codon usage for each full-length protein gene has been calculated using the nucleotide sequence obtained from GenBank sequence database. The sum of the codon use of each organism has been also calculated. The data files can be obtained from anonymous ftp sites of DDBJ, DISC and EBI. The list of codonusage of genes in organisms was made searchableby name of organism through a web site http://www.dna.affrc.go.jp/ approximately nakamura/CUTG.html The compilation is synchronized with major release of GenBank. PMID- 9399869 TI - The translational signal database, TransTerm, is now a relational database. AB - TransTerm-97 contains more than 97 500 non-redundant coding-sequence initiation and termination contexts compiled from GenBank, release 101 (15-June-1997). In addition, several coding sequence parameters are available: coding sequence length, Nc, GC3, and, when it is computable, codon adaptation index (CAI). Codon usage tables and summaries of start and stop codon contexts are also included. The information covers more than 325 species and organelles, including seven complete bacterial genomes and one complete eukaryotic genome. To promote research in translational control of protein synthesis, TransTerm has been converted into a relational database to ease the process of making queries. The relational database manager, Postgresql, gives access to the database using SQL (Structured Query Language). A World Wide Web interface using forms is being completed to allow the casual user access to the database. Extensions are planned to include the full 5'-UTR, full coding sequence and 3'-UTR. TransTerm-97 is available on the World Wide Web at:http://biochem. otago.ac.nz:800/Transterm/homepage.html PMID- 9399870 TI - REBASE - restriction enzymes and methylases. AB - REBASE is a comprehensive database of information about restriction enzymes and their associated methylases, including their recognition and cleavage sites and their commercial availability. Also included is a listing of homing endonucleases. Information from REBASE is available via monthly electronic mailings as well as via anonymous ftp and through the World Wide Web. The REBASE web site, http://www. neb.com/rebase , is where we maintain a web page for every enzyme, reference and supplier. Additionally, there is a search facility, help and NEWS pages, and a complete description of our various services. Specialized files are available that can be used directly by many software packages. PMID- 9399871 TI - The ribonuclease P database. AB - Ribonuclease P is responsible for the 5'-maturation of tRNA precursors. Ribonuclease P is a ribonucleoprotein, and in bacteria the RNA subunit alone is catalytically active in vitro , i.e., it is a ribozyme. The Ribonuclease P Database is a compilation of ribonuclease P sequences, sequence alignments, secondary structures, three-dimensional models, and accessory information, available via the World Wide Web (http: //www.mbio.ncsu.edu/RNaseP/home.html ). PMID- 9399872 TI - The Eukaryotic Promoter Database EPD. AB - The Eukaryotic Promoter Database (EPD) is an annotated non-redundant collection of experimentally characterised eukaryotic POL II promoters. The underlying definition of a promoter is that of a transcription initiation site. All information presented in EPD results from an independent evaluation of primary experimental data shown in the biological literature. Sequences flanking transcription initiation sites are indirectly given by pointers to EMBL sequences. The annotation part of a promoter entry includes description of the promoter-defining evidence, cross-references to other databases, and bibliographic references. Being designed as a resource for comparative sequence analysis, EPD is structured in a way that facilitates dynamic extraction of biologically meaningful promoter subsets. The database is available through the World Wide Web at URL http://cmpteam4.unil.ch PMID- 9399873 TI - PLACE: a database of plant cis-acting regulatory DNA elements. AB - PLACE (http://www.dna.affrc.go.jp/htdocs/PLACE/) is a database of motifs found in plant cis -acting regulatory DNA elements, all from previously published reports. It covers vascular plants only. In addition to the motifs originally reported, their variations in other genes or in other plant species reported later are also compiled. The PLACE database also contains a brief description of each motif and relevant literature with PubMed ID numbers. This report summarizes the present status of this database and available tools. PMID- 9399874 TI - OOTFD (Object-Oriented Transcription Factors Database): an object-oriented successor to TFD. AB - ooTFD (object-oriented Transcription Factors Database) is a successor to TFD (Transcription Factors Database). ooTFD contains information represented in TFD but also allows the representation of containment, composite, and interaction relationships between transcription factor polypeptides. ooTFD is designed to represent information about all transcription factors, both eukaryotic and prokaryotic, basal as well as regulatory factors, and multiprotein complexes as well as monomers. ooTFD and associated tools and services can be accessed at http://www.isbi.net/ PMID- 9399875 TI - Databases on transcriptional regulation: TRANSFAC, TRRD and COMPEL. AB - TRANSFAC, TRRD (Transcription Regulatory Region Database) and COMPEL are databases which store information about transcriptional regulation in eukaryotic cells. The three databases provide distinct views on the components involved in transcription: transcription factors and their binding sites and binding profiles (TRANSFAC), the regulatory hierarchy of whole genes (TRRD), and the structural and functional properties of composite elements (COMPEL). The quantitative and qualitative changes of all three databases and connected programs are described. The databases are accessible via WWW:http://transfac.gbf.de/TRANSFAC orhttp://www.bionet.nsc.ru/TRRD PMID- 9399876 TI - MHCPEP, a database of MHC-binding peptides: update 1997. AB - MHCPEP (http://wehih.wehi.edu.au/mhcpep/) is a curated database comprising over 13 000 peptide sequences known to bind MHC molecules. Entries are compiled from published reports as well as from direct submissions of experimental data. Each entry contains the peptide sequence, its MHC specificity and where available, experimental method, observed activity, binding affinity, source protein and anchor positions, as well as publication references. The present format of the database allows text string matching searches but can easily be converted for use in conjunction with sequence analysis packages. The database can be accessed via Internet using WWW or FTP. PMID- 9399877 TI - Histone Sequence Database: new histone fold family members. AB - Searches of the major public protein databases with core and linker chicken and human histone sequences have resulted in the compilation of an annotated set of histone protein sequences. In addition, new database searches with two distinct motif search algorithms have identified several members of the histone fold family, including human DRAP1 and yeast CSE4. Database resources include information on conflicts between similar sequence entries in different source databases, multiple sequence alignments, links to the Entrez integrated information retrieval system, structures for histone and histone fold proteins, and the ability to visualize structural data through Cn3D. The database currently contains >1000 protein sequences, which are searchable by protein type, accession number, organism name, or any other free text appearing in the definition line of the entry. All sequences and alignments in this database are available through the World Wide Web at http://www.nhgri.nih. gov/DIR/GTB/HISTONES or http://www.ncbi.nlm.nih. gov/Baxevani/HISTONES PMID- 9399878 TI - PROMISE: a database of information on prosthetic centres and metal ions in protein active sites. AB - The PROMISE (Prosthetic centres andmetalions in protein activesites) database aims to gather together comprehensive sequence, structural, functional and bibliographic information on proteins which possess prosthetic centres, with an emphasis on active site structure and function. The database is available on the World Wide Web at http://bioinf.leeds.ac.uk/promise/ PMID- 9399879 TI - PhosphoBase: a database of phosphorylation sites. AB - PhosphoBase is a database of experimentally verified phosphorylation sites. Version 1.0 contains 156 entries and 398 experimentally determined phosphorylation sites. Entries are compiled and revised from the literature and from major protein sequence databases such as SwissProt and PIR. The entries provide information about the phosphoprotein and the exact position of its phosphorylation sites. Furthermore, part of the entries contain information about kinetic data obtained from enzyme assays on specific peptides. To illustrate the use of data extracted from PhosphoBase we present a sequence logo displaying the overall conservation of positions around serines phosphorylated by protein kinase A (PKA). PhosphoBase is available on the WWW at http://www.cbs.dtu.dk/databases/PhosphoBase/ PMID- 9399880 TI - O-GLYCBASE Version 3.0: a revised database of O-glycosylated proteins. AB - O-GLYCBASE is a revised database of information on glycoproteins and their O linked glycosylation sites. Entries are compiled and revised from the literature, and from the sequence databases. Entries include information about species, sequence, glycosylation sites and glycan type and is fully cross-referenced. Compared to version 2.0 the number of entries has increased by 20%. Sequence logos displaying the acceptor specificity patterns for the GalNAc, mannose and GlcNAc transferases are shown. The O-GLYCBASE database is available through the WWW at http://www.cbs.dtu. dk/databases/OGLYCBASE/ PMID- 9399881 TI - Use of a quantitative trait to map a locus associated with severity of positive symptoms in familial schizophrenia to chromosome 6p. AB - A number of recent linkage studies have suggested the presence of a schizophrenia susceptibility locus on chromosome 6p. We evaluated 28 genetic markers, spanning chromosome 6, for linkage to schizophrenia in 10 moderately large Canadian families of Celtic ancestry. Parametric analyses of these families under autosomal dominant and recessive models, using broad and narrow definitions of schizophrenia, produced no significant evidence for linkage. A sib-pair analysis using categorical disease definitions also failed to produce significant evidence for linkage. We then conducted a separate sibpair analysis using scores on positive-symptom (psychotic), negative-symptom (deficit), and general psychopathology-symptom scales as quantitative traits. With the positive symptom scale scores, the marker D6S1960 produced P = 1.2 x 10(-5) under two-point and P = 5.4 x 10(-6) under multipoint analyses. Using simulation studies, we determined that these nominal P values correspond to empirical P values of .034 and .0085, respectively. These results suggest that a schizophrenia susceptibility locus on chromosome 6p may be related to the severity of psychotic symptoms. Assessment of behavioral quantitative traits may provide increased power over categorical phenotype assignment for detection of linkage in complex psychiatric disorders. PMID- 9399882 TI - Inherited interstitial duplications of proximal 15q: genotype-phenotype correlations. AB - We present the cytogenetic, molecular cytogenetic, and molecular genetic results on 20 unrelated patients with an interstitial duplication of the proximal long arm of chromosome 15. Multiple probes showed that the Prader-Willi/Angelman critical region (PWACR) was included in the duplication in 4/20 patients, each ascertained with developmental delay. The duplication was also found in two affected but not in three unaffected sibs of one of these patients. All four probands had inherited their duplication from their mothers, three of whom were also affected. Two of the affected mothers also carried a maternally inherited duplication, whereas the duplication in the unaffected mother and in an unaffected grandmother was paternal in origin, raising the possibility of a parental-origin effect. The PWACR was not duplicated in the remaining 16 patients, of whom 4 were referred with developmental delay. In the 14 families for which parental samples were available, the duplication was inherited with equal frequency from a phenotypically normal parent, mother or father. Comparative genomic hybridization undertaken on two patients suggested that proximal 15q outside the PWACR was the origin of the duplicated material. The use of PWACR probes discriminates between a large group of duplications of no apparent clinical significance and a smaller group, in which a maternally derived PWACR duplication is consistently associated with developmental delay and speech difficulties but not with overt features of either Prader-Willi syndrome or Angelman syndrome. PMID- 9399885 TI - Genetic segregation analysis of early-onset recurrent unipolar depression. AB - Major depression is a relatively common psychiatric disorder that can be quite debilitating. Family, twin, and adoption studies indicate that unipolar depression has both genetic and environmental components. Early age at onset and recurrent episodes in the proband each increase the familiarity of the illness. To investigate the potential genetic underpinnings of the disease, we have performed a complex segregation analysis on 832 individuals from 50 multigenerational families ascertained through a proband with early-onset recurrent unipolar major depression. The analysis was conducted by use of regressive models, to test a variety of hypotheses to explain the familial aggregation of recurrent unipolar depression. Analyses were conducted under two alternative definitions of affection status for the relatives of probands: (1) "narrow," in which relatives were assumed to be affected only if they were diagnosed with recurrent unipolar depression; and (2) "broad," in which relatives were assumed to be affected if diagnosed with any major affective illness. Under the narrow-definition assumption, the model that best explains these family data is a transmitted (although non-Mendelian) recessive major effect with significant residual parental effects on affection status. Under the broad-definition assumption, the best-fitting model is a Mendelian codominant major locus with significant residual parental and spousal effects. PMID- 9399886 TI - Cloning of the human carnitine-acylcarnitine carrier cDNA and identification of the molecular defect in a patient. AB - The carnitine-acylcarnitine carrier (CAC) catalyzes the translocation of long chain fatty acids across the inner mitochondrial membrane. We cloned and sequenced the human CAC cDNA, which has an open reading frame of 903 nucleotides. Northern blot studies revealed different expression levels of CAC in various human tissues. Furthermore, mutation analysis was performed for a CAC-deficient infant. Direct sequencing of the patient's cDNA revealed a homozygous cytosine nucleotide insertion. This insertion provokes a frameshift and an extension of the open reading frame with 23 novel codons. This is the first report documenting a mutation, in the CAC cDNA, responsible for mitochondrial beta-oxidation impairment. PMID- 9399887 TI - Diversity and age of the four major mtDNA haplogroups, and their implications for the peopling of the New World. AB - Despite considerable investigation, two main questions on the origin of Native Americans remain the topic of intense debate-namely, the number and time of the migration(s) into the Americas. Using the 720 available Amerindian mtDNA control region sequences, we reanalyzed the nucleotide diversity found within each of the four major mtDNA haplogroups (A-D) thought to have been present in the colonization of the New World. We first verified whether the within-haplogroup sequence diversity could be used as a measure of the haplogroup's age. The pattern of shared polymorphism, the mismatch distribution, the phylogenetic trees, the value of Tajima's D, and the computer simulations all suggested that the four haplogroups underwent a bottleneck followed by a large population expansion. The four haplogroup diversities were very similar to each other, offering a strong support for their single origin. They suggested that the beginning of the Native Americans' ancestral-population differentiation occurred approximately 30,000-40,000 years before the present (ybp), with a 95%-confidence interval lower bound of approximately 25,000 ybp. These values are in good agreement with the New World-settlement model that we have presented elsewhere, extending the results initially found for haplogroup A to the three other major groups of mtDNA sequences found in the Americas. These results put the peopling of the Americas clearly in an early, pre-Clovis time frame. PMID- 9399888 TI - Linkage of bipolar affective disorder to chromosome 18 markers in a new pedigree series. AB - Several groups have reported evidence suggesting linkage of bipolar affective disorder (BPAD) to chromosome 18. We have reported data from 28 pedigrees that showed linkage to marker loci on 18p and to loci 40 cM distant on 18q. Most of the linkage evidence derived from families with affected phenotypes in only the paternal lineage and from marker alleles transmitted on the paternal chromosome. We now report results from a series of 30 new pedigrees (259 individuals) genotyped for 13 polymorphic markers spanning chromosome 18. Subjects were interviewed by a psychiatrist and were diagnosed by highly reliable methods. Genotypes were generated with automated technology and were scored blind to phenotype. Affected sib pairs showed excess allele sharing at the 18q markers D18S541 and D18S38. A parent-of-origin effect was observed, but it was not consistently paternal. No robust evidence of linkage was detected for markers elsewhere on chromosome 18. Multipoint nonparametric linkage analysis in the new sample combined with the original sample of families supports linkage on chromosome 18q, but the susceptibility gene is not well localized. PMID- 9399889 TI - Meiotic microdeletion breakpoints in the BRCA1 gene are significantly associated with symmetric DNA-sequence elements. PMID- 9399890 TI - Mutations of the Fanconi anemia group A gene (FAA) in Italian patients. AB - Fanconi anemia (FA) is an autosomal recessive disease characterized by progressive pancytopenia, congenital malformations, and predisposition to acute myeloid leukemia. At least five complementation groups (FA-A-FA-E) have been identified. The relative prevalence of FA-A has been estimated at an average of approximately 65% but may widely vary according to ethnic background. In Italy, 11 of 12 patients analyzed by cell-fusion studies were assigned to group FA-A, suggesting an unusually high relative prevalence of this FA subtype in patients of Italian ancestry. We have screened the 43 exons of the FAA gene and their flanking intronic sequences in 38 Italian FA patients, using RNA-SSCP. Ten different mutations were detected: three nonsense and one missense substitutions, four putative splice mutations, an insertion, and a duplication. Most of the mutations are expected to cause a premature termination of the FAA protein at various sites throughout the molecule. Four protein variants were also found, three of which were polymorphisms. The missense mutation D1359Y, not found in chromosomes from healthy unrelated individuals, was responsible for a local alteration of hydrophobicity in the FAA protein, and it was likely to be pathogenic. Thus, the mutations so far encountered in the FAA gene are essentially all different. Since screening based on the analysis of single exons by genomic DNA amplification apparently detects only a minority of the mutations, methods designed to detect alterations in the genomic structure of the gene or in the FAA polypeptide may be helpful in the identification of FAA mutations. PMID- 9399892 TI - Identification of an interstitial deletion in an adult female with schizophrenia, mental retardation, and dysmorphic features: further support for a putative schizophrenia-susceptibility locus at 5q21-23.1. PMID- 9399891 TI - Molecular analysis of the NF2 tumor-suppressor gene in schwannomatosis. AB - Patients with multiple schwannomas without vestibular schwannomas have been postulated to compose a distinct subclass of neurofibromatosis (NF), termed "schwannomatosis." To compare the molecular-genetic basis of schwannomatosis with NF2, we examined the NF2 locus in 20 unrelated schwannomatosis patients and their affected relatives. Tumors from these patients frequently harbored typical truncating mutations of the NF2 gene and loss of heterozygosity of the surrounding region of chromosome 22. Surprisingly, unlike patients with NF2, no heterozygous NF2-gene changes were seen in normal tissues. Examination of multiple tumors from the same patient revealed that some schwannomatosis patients are somatic mosaics for NF2-gene changes. By contrast, other individuals, particularly those with a positive family history, appear to have an inherited predisposition to formation of tumors that carry somatic alterations of the NF2 gene. Further work is needed to define the pathogenetics of this unusual disease mechanism. PMID- 9399893 TI - Comparison of nonparametric statistics for detection of linkage in nuclear families: single-marker evaluation. AB - We have evaluated 23 different statistics, from a total of 10 popular software packages for model-free linkage analysis of nuclear-family data, by applying them to single-marker data simulated under several two-locus disease models. The statistics that we examined fall into two broad categories: (1) those that test directly for increased identity-by-state or identity-by-descent sharing (by use of the programs APM, Genetic Analysis System [GAS] SIBSTATE and SIBDES, SAGE SIBPAL, ERPA, SimIBD, and Genehunter NPL) and (2) those that are based on likelihood-ratio tests and that report LOD scores (by use of the programs Splink, SIBPAIR, Mapmaker/Sibs, ASPEX, and GAS SIBMLS). For each of eight two-locus disease models, we analyzed six data sets; the first three data sets consisted of two-child families with both sibs affected and zero, one, or both parents typed, whereas the other three data sets consisted of four-child families with at least two affected sibs and zero, one, or both parents typed. We report false-positive rates, overall rank by power, and the power for each statistic. We give rough recommendations regarding which programs provide the most powerful tests for linkage, as well as the programs to be avoided under certain conditions. For the likelihood-ratio-based statistics, we examined the effects of various treatments of sibships with multiple affected individuals. Finally, we explored the use of some simple two-of-three composite statistics and found that such tests are of only marginal benefit over the most powerful single statistic. PMID- 9399894 TI - Variable levels of a heteroplasmic point mutation in individual hair roots. AB - During direct sequencing of the first hypervariable segment of the human mitochondrial control region, we identified one individual with a heteroplasmic point mutation at nt 16,256. We used primer extension to analyze the proportions of each mitochondrial haplotype in peripheral blood, buccal cells, and single hair roots from this individual and from eight members of his maternal lineage. Significant levels of heteroplasmy were found in only three individuals, and, in these cases, the proportions of each haplotype were similar in both blood and buccal cells. From the changes in mitochondrial haplotypes within mother offspring pairs, we calculated that the most likely size of a mitochondrial bottleneck during development was 1-27 segregating units. However, highly variable levels of heteroplasmy were found in single hair roots, even among roots from the same individual. We analyzed a large number of hair roots from one individual and found that the proportion of one haplotype was within a range of 9% to > 99% in different roots. Roots originating from within a small patch of skin had haplotype proportions as variable as those from different areas of skin. PMID- 9399895 TI - Genomewide scan of multiple sclerosis in Finnish multiplex families. AB - Multiple sclerosis (MS) is a neurological, demyelinating disorder with a putative autoimmune etiology. It is thought to be a multifactorial disease with a complex mode of inheritance. Here we report the results of a two-stage genomewide scan for loci predisposing to MS. The first stage of the screen, with a low-resolution map, was performed in a selection of 16 pedigrees collected from an isolated Finnish population. Multipoint, non-parametric linkage analysis of the 328 markers did not reveal statistically significant results. However, 10 slightly interesting regions (P = .1-.15) emerged, including our previous findings of the HLA complex on 6p21 and a putative locus on 5p14-p12. Eight of these novel regions were further analyzed by use of denser marker maps, in the second stage of the scan. For the chromosomal regions 4cen, 11tel, and 17q, the statistical significance increased, but not conclusively; for 2q32 and 10q21, the statistical significance did not change. Accordingly, genotyping of the high-density markers in these regions was performed, and the data were analyzed by use of two-point, parametric linkage analysis using the complete pedigree information of the 21 Finnish multiplex families. We detected suggestive evidence for a predisposing locus on chromosomal region 17q22-q24. Several markers on 17q22-q24 yielded positive LOD scores, with the maximum LOD score (Zmax) occurring with D17S807 (Zmax = 2.8, theta = .04; dominant model). Interestingly, a suggestive linkage between MS and the markers on 17q22-q24 was also revealed by a recent genomewide scan in MS families from the United Kingdom. PMID- 9399896 TI - Human phenylalanine hydroxylase mutations and hyperphenylalaninemia phenotypes: a metanalysis of genotype-phenotype correlations. AB - We analyzed correlations between mutant genotypes at the human phenylalanine hydroxylase locus (gene symbol PAH) and the corresponding hyperphenylalaninemia (HPA) phenotypes (notably, phenylketonuria [OMIM 261600]). We used reports, both published and in the PAH Mutation Analysis Consortium Database, on 365 patients harboring 73 different PAH mutations in 161 different genotypes. HPA phenotypes were classified as phenylketonuria (PKU), variant PKU, and non-PKU HPA. By analysis both of homoallelic mutant genotypes and of "functionally hemizygous" heteroallelic genotypes, we characterized the phenotypic effect of 48 of the 73 different, largely missense mutations. Among those with consistent in vivo expression, 24 caused PKU, 3 caused variant PKU, and 10 caused non-PKU HPA. However, 11 mutations were inconsistent in their effect: 9 appeared in two different phenotype classes, and 2 (I65T and Y414C) appeared in all three classes. Seven mutations were inconsistent in phenotypic effect when in vitro (unit-protein) expression was compared with the corresponding in vivo phenotype (an emergent property). We conclude that the majority of PAH mutations confer a consistent phenotype and that this is concordant with their effects, when known, predicted from in vitro expression analysis. However, significant inconsistencies, both between in vitro and in vivo phenotypes and between different individuals with similar PAH genotypes, reveal that the HPA-phenotype is more complex than that predicted by Mendelian inheritance of alleles at the PAH locus. PMID- 9399898 TI - The frequency of the methylenetetrahydrofolate reductase-gene mutation varies with age in the normal population. PMID- 9399897 TI - Inherited mutations in PTEN that are associated with breast cancer, cowden disease, and juvenile polyposis. AB - PTEN, a protein tyrosine phosphatase with homology to tensin, is a tumor suppressor gene on chromosome 10q23. Somatic mutations in PTEN occur in multiple tumors, most markedly glioblastomas. Germ-line mutations in PTEN are responsible for Cowden disease (CD), a rare autosomal dominant multiple-hamartoma syndrome. PTEN was sequenced from constitutional DNA from 25 families. Germ-line PTEN mutations were detected in all of five families with both breast cancer and CD, in one family with juvenile polyposis syndrome, and in one of four families with breast and thyroid tumors. In this last case, signs of CD were subtle and were diagnosed only in the context of mutation analysis. PTEN mutations were not detected in 13 families at high risk of breast and/or ovarian cancer. No PTEN coding-sequence polymorphisms were detected in 70 independent chromosomes. Seven PTEN germ-line mutations occurred, five nonsense and two missense mutations, in six of nine PTEN exons. The wild-type PTEN allele was lost from renal, uterine, breast, and thyroid tumors from a single patient. Loss of PTEN expression was an early event, reflected in loss of the wild-type allele in DNA from normal tissue adjacent to the breast and thyroid tumors. In RNA from normal tissues from three families, mutant transcripts appeared unstable. Germ-line PTEN mutations predispose to breast cancer in association with CD, although the signs of CD may be subtle. PMID- 9399899 TI - Splicing defects in the COL3A1 gene: marked preference for 5' (donor) spice-site mutations in patients with exon-skipping mutations and Ehlers-Danlos syndrome type IV. AB - Ehlers-Danlos syndrome (EDS) type IV results from mutations in the COL3A1 gene, which encodes the constituent chains of type III procollagen. We have identified, in 33 unrelated individuals or families with EDS type IV, mutations that affect splicing, of which 30 are point mutations at splice junctions and 3 are small deletions that remove splice-junction sequences and partial exon sequences. Except for one point mutation at a donor site, which leads to partial intron inclusion, and a single base-pair substitution at an acceptor site, which gives rise to inclusion of the complete upstream intron into the mature mRNA, all mutations result in deletion of a single exon as the only splice alteration. Of the exon-skipping mutations that are due to single base substitutions, which we have identified in 28 separate individuals, only two affect the splice-acceptor site. The underrepresentation of splice acceptor-site mutations suggests that the favored consequence of 3' mutations is the use of an alternative acceptor site that creates a null allele with a premature-termination codon. The phenotypes of those mutations may differ, with respect to either their severity or their symptomatic range, from the usual presentation of EDS type IV and thus have been excluded from analysis. PMID- 9399900 TI - Localization of the gene for thiamine-responsive megaloblastic anemia syndrome, on the long arm of chromosome 1, by homozygosity mapping. AB - Thiamine-responsive megaloblastic anemia, also known as "TRMA" or "Rogers syndrome," is an early-onset autosomal recessive disorder defined by the occurrence of megaloblastic anemia, diabetes mellitus, and sensorineural deafness, responding in varying degrees to thiamine treatment. On the basis of a linkage analysis of affected families of Alaskan and of Italian origin, we found, using homozygosity mapping, that the TRMA-syndrome gene maps to a region on chromosome 1q23.2-23.3 (maximum LOD score of 3.7 for D1S1679). By use of additional consanguineous kindreds of Israeli-Arab origin, the putative disease gene interval also has been confirmed and narrowed, suggesting genetic homogeneity. Linkage analysis generated the highest combined LOD-score value, 8.1 at a recombination fraction of 0, with marker D1S2799. Haplotype analysis and recombination events narrowed the TRMA locus to a 16-cM region between markers D1S194 and D1S2786. Several heterozygote parents had diabetes mellitus, deafness, or megaloblastic anemia, which raised the possibility that mutations at this locus predispose carriers in general to these manifestations. Characterization of the metabolic defect of TRMA may shed light on the role of thiamine deficiency in such common diseases. PMID- 9399901 TI - Autosomal dominant postaxial polydactyly, nail dystrophy, and dental abnormalities map to chromosome 4p16, in the region containing the Ellis-van Creveld syndrome locus. AB - We have studied a four-generation family with features of Weyers acrofacial dysostosis, in which the proband has a more severe phenotype, resembling Ellis van Creveld syndrome. Weyers acrofacial dysostosis is an autosomal dominant condition with dental anomalies, nail dystrophy, postaxial polydactyly, and mild short stature. Ellis-van Creveld syndrome is a similar condition, with autosomal recessive inheritance and the additional features of disproportionate dwarfism, thoracic dysplasia, and congenital heart disease. Linkage and haplotype analysis determined that the disease locus in this pedigree resides on chromosome 4p16, distal to the genetic marker D4S3007 and within a 17-cM region flanking the genetic locus D4S2366. This region includes the Ellis-van Creveld syndrome locus, which previously was reported to map within a 3-cM region between genetic markers D4S2957 and D4S827. Either the genes for the condition in our family and for Ellis-van Creveld syndrome are near one another or these two conditions are allelic with mutations in the same gene. These data also raise the possibility that Weyers acrofacial dysostosis is the heterozygous expression of a mutation that, in homozygous form, causes the autosomal recessive disorder Ellis-van Creveld syndrome. PMID- 9399902 TI - Peutz-Jeghers syndrome: confirmation of linkage to chromosome 19p13.3 and identification of a potential second locus, on 19q13.4. AB - Peutz-Jeghers syndrome (PJS) is an autosomal dominant disease with variable expression and incomplete penetrance, characterized by mucocutaneous pigmentation and hamartomatous polyposis. Patients with PJS have increased frequency of gastrointestinal and extraintestinal malignancies (ovaries, testes, and breast). In order to map the locus (or loci) associated with PJS, we performed a genomewide linkage analysis, using DNA polymorphisms in six families (two from Spain, two from India, one from the United States, and one from Portugal) comprising a total of 93 individuals, including 39 affected and 48 unaffected individuals and 6 individuals with unknown status. During this study, localization of a PJS gene to 19p13.3 (around marker D19S886) had been reported elsewhere. For our families, marker D19S886 yielded a maximum LOD score of 4.74 at a recombination fraction (theta) of .045; multipoint linkage analysis resulted in a LOD score of 7.51 for the interval between D19S886 and 19 pter. However, markers on 19q13.4 also showed significant evidence for linkage. For example, D19S880 resulted in a maximum LOD score of 3.8 at theta = .13. Most of this positive linkage was contributed by a single family, PJS07. These results confirm the mapping of a common PJS locus on 19p13.3 but also suggest the existence, in a minority of families, of a potential second PJS locus, on 19q13.4. Positional cloning and characterization of the PJS mutations will clarify the genetics of the syndrome and the implication of the gene(s) in the predisposition to neoplasias. PMID- 9399904 TI - Spectrum of mutations in the RPGR gene that are identified in 20% of families with X-linked retinitis pigmentosa. AB - The RPGR (retinitis pigmentosa GTPase regulator) gene for RP3, the most frequent genetic subtype of X-linked retinitis pigmentosa (XLRP), has been shown to be mutated in 10%-15% of European XLRP patients. We have examined the RPGR gene for mutations in a cohort of 80 affected males from apparently unrelated XLRP families, by direct sequencing of the PCR-amplified products from the genomic DNA. Fifteen different putative disease-causing mutations were identified in 17 of the 80 families; these include four nonsense mutations, one missense mutation, six microdeletions, and four intronic-sequence substitutions resulting in splice defects. Most of the mutations were detected in the conserved N-terminal region of the RPGR protein, containing tandem repeats homologous to those present in the RCC-1 protein (a guanine nucleotide-exchange factor for Ran-GTPase). Our results indicate that mutations either in as yet uncharacterized sequences of the RPGR gene or in another gene located in its vicinity may be a more frequent cause of XLRP. The reported studies will be beneficial in establishing genotype-phenotype correlations and should lead to further investigations seeking to understand the mechanism of disease pathogenesis. PMID- 9399903 TI - Novel alleles of the chemokine-receptor gene CCR5. AB - The CCR5 gene encodes a cell-surface chemokine-receptor molecule that serves as a coreceptor for macrophage-tropic strains of HIV-1. Mutations in this gene may alter expression or function of the protein product, thereby altering chemokine binding/signaling or HIV-1 infection of cells that normally express CCR5 protein. Indeed, homozygotes for a 32-bp deletion allele of CCR5 (CCR5-delta 32), which causes a frameshift at amino acid 185, are relatively resistant to HIV-1 infection. Here we report the identification of 16 additional mutations in the coding region of the CCR5 gene, all but 3 of which are codon altering or "nonsynonymous." Most mutations were rare (found only once or twice in the sample); five were detected exclusively among African Americans, whereas eight were observed only in Caucasians. The mutations included 11 codon-altering nonsynonymous variants, one trinucleotide deletion, one chain-termination mutant, and three synonymous mutations. The high predominance of codon-altering alleles among CCR5 mutants (14/17 [81%], including CCR5-delta 32) is consistent with an adaptive accumulation of function-altering alleles for this gene, perhaps as a consequence of historic selective pressures. PMID- 9399905 TI - Fragile X premutations are not a major cause of early menopause. AB - Fragile X syndrome is an X-linked mental retardation condition that usually is due to a trinucleotide-repeat expansion in the FMR1 gene. Whereas full-mutation alleles (> 230 repeats) lead to fragile X syndrome, premutation alleles (approximately 60-200 repeats) are apparently non-penetrant. However, previous studies have suggested that female premutation carriers may have an increased incidence of premature menopause. To test this possible association, we screened for premutation alleles among 216 women with early menopause (at age < 47 years), 33 of whom had premature menopause (at age < 40 years), as well as among 107 control women, all of whom were ascertained solely on the basis of age at menopause. No full-mutation alleles were found; and only one premutation allele was found, but, it was in a member of the control group. These results are consistent with what would be expected on the basis of chance only. Our sample size was sufficient to rule out a > or = 3-fold increased risk of early menopause and a > or = 9-fold increased risk of premature menopause due to an FMR1 premutation, under a model considering the risk of both sporadic and familial early menopause. Likewise, our results rule out a > or = 4-fold increased risk of familial early menopause and a > or = 26-fold increased risk of familial premature menopause, under a less probable model in which only familial early menopause is considered. These results indicate that the fragile X premutation is not a major risk factor for early menopause and suggest that the risk of premature menopause to fragile X-premutation carriers may not be as great as that reported elsewhere. PMID- 9399906 TI - Genomewide transmission/disequilibrium testing--consideration of the genotypic relative risks at disease loci. AB - Genomewide association studies are set to become the tool of the future for detection of small-effect genes in complex diseases. It will therefore be necessary to calculate sufficient sample sizes with which to perform them. In this paper I illustrate how to calculate the required number of families for general genotypic relative risks (GRRs). I show the superior sensitivity of the genomewide association study over the standard genomewide affected-sib-pair linkage analysis, for a range of different underlying GRR patterns. I also illustrate the extent of change in the sample sizes that is necessary for a genomewide association analysis depending on the pattern of the GRRs at the disease locus. In many cases, the comparative numbers of families required under different genetic mechanisms vary by several orders of magnitude. These sometimes dramatic differences have important implications for the determination of the size of the collection of samples prior to analysis and for the types of effects that are likely--and unlikely--to be detected by such an analysis. PMID- 9399907 TI - Homogeneity of kerato-epithelin codon 124 mutations in Japanese patients with either of two types of corneal stromal dystrophy. PMID- 9399908 TI - Isolation and chromosomal localization of a cornea-specific human keratin 12 gene and detection of four mutations in Meesmann corneal epithelial dystrophy. AB - Keratin 12 (K12) is an intermediate-filament protein expressed specifically in corneal epithelium. Recently, we isolated K12 cDNA from a human corneal epithelial cDNA library and determined its full sequence. Herein, we present the exon-intron boundary structure and chromosomal localization of human K12. In addition, we report four K12 mutations in Meesmann corneal epithelial dystrophy (MCD), an autosomal dominant disorder characterized by intraepithelial microcysts and corneal epithelial fragility in which mutations in keratin 3 (K3) and K12 have recently been implicated. In the human K12 gene, we identified seven introns, defining eight individual exons that cover the coding sequence. Together the exons and introns span approximately 6 kb of genomic DNA. Using FISH, we found that the K12 gene mapped to 17q12, where a type I keratin cluster exists. In this study, four new K12 mutations (Arg135Gly, Arg135Ile, Tyr429Asp, and Leu140Arg) were identified in three unrelated MCD pedigrees and in one individual with MCD. All mutations were either in the highly conserved alpha-helix initiation motif of rod domain 1A or in the alpha-helix-termination motif of rod domain 2B. These sites are essential for keratin filament assembly, suggesting that the mutations described above may be causative for MCD. Of particular interest, one of these mutations (Tyr429Asp), detected in both affected individuals in one of our pedigrees, is the first mutation to be identified within the alpha-helix-termination motif in type I keratin. PMID- 9399909 TI - Skewed X-chromosome inactivation is common in fetuses or newborns associated with confined placental mosaicism. AB - The inactivation of one X chromosome in females is normally random with regard to which X is inactivated. However, exclusive or almost-exclusive inactivation of one X may be observed in association with some X-autosomal rearrangements, mutations of the XIST gene, certain X-linked diseases, and MZ twinning. In the present study, a methylation difference near a polymorphism in the X-linked androgen-receptor gene was used to investigate the possibility that nonrandom X inactivation is increases in fetuses and newborns that are associated with confined placental mosaicism (CPM) involving an autosomal trisomy. Extreme skewing was observed in 7 (58%) of 12 cases with a meiotic origin of the trisomy, but in none of 10 cases examined with a somatic origin of the trisomy, and in only 1 (4%) of 27 control adult females. In addition, an extremely skewed X inactivation pattern was observed in 3 of 10 informative cases of female uniparental disomy (UPD) of chromosome 15. This may reflect the fact that a proportion of UPD cases arise by "rescue" of a chromosomally abnormal conceptus and are therefore associated with CPM. A skewed pattern of X inactivation in CPM cases is hypothesized to result from a reduction in the size of the early embryonic cell pool, because of either poor early growth or subsequent selection against the trisomic cells. Since approximately 2% of pregnancies detected by chorionic villus sampling are associated with CPM, this is likely a significant contributor to both skewed X inactivation observed in the newborn population and the expression of recessive X-linked diseases in females. PMID- 9399910 TI - Goosecoid-like sequences and the smallest region of deletion overlap in DiGeorge and velocardiofacial syndromes. PMID- 9399912 TI - Instability of the (CTG)n repeat in congenital myotonic dystrophy. PMID- 9399911 TI - Mutations in PDX1, the human lipoyl-containing component X of the pyruvate dehydrogenase-complex gene on chromosome 11p1, in congenital lactic acidosis. AB - We have identified and sequenced a cDNA that encodes an apparent human orthologue of a yeast protein-X component (ScPDX1) of pyruvate dehydrogenase multienzyme complexes. The new human cDNA that has been referred to as "HsPDX1" cDNA was cloned by use of the "database cloning" strategy and had a 1,506-bp open reading frame. The amino acid sequence of the protein encoded by the cDNA was 20% identical with that encoded by the yeast PDX1 gene and 40% identical with that encoded by the lipoate acetyltransferase component of the pyruvate dehydrogenase and included a lipoyl-bearing domain that is conserved in some dehydrogenase enzyme complexes. Northern blot analysis demonstrated that the major HsPDX1 mRNA was 2.5 kb in length and was expressed mainly in human skeletal and cardiac muscles but was also present, at low levels, in other tissues. FISH analysis performed with a P1-derived artificial chromosome (PAC)-containing HsPDX1 gene sublocalized the gene to 11p1.3. Molecular investigation of PDX1 deficiency in four patients with neonatal lactic acidemias revealed mutations 78del85 and 965del59 in a homozygous state, and one other patient had no PDX1 mRNA expression. PMID- 9399913 TI - Genomic imprinting: a chromatin connection. PMID- 9399915 TI - Centromere DNA dynamics: latent centromeres and neocentromere formation. PMID- 9399916 TI - Dynamic interrelationships between DNA replication, methylation, and repair. PMID- 9399935 TI - Clinically relevant findings. PMID- 9399917 TI - PTEN: sometimes taking it off can be better than putting it on. PMID- 9399936 TI - An emerging paradigm shift on the role of leukocyte adhesion molecules. PMID- 9399937 TI - The Epstein-Barr virus and systemic lupus erythematosus. PMID- 9399938 TI - Pathways and mechanisms for cytokine signaling of the central nervous system. PMID- 9399939 TI - Cytokines in the brain during viral infection: clues to HIV-associated dementia. PMID- 9399940 TI - Aberrant nuclear factor-kappaB/Rel expression and the pathogenesis of breast cancer. AB - Expression of nuclear factor-kappaB (NF-kappaB)/Rel transcription factors has recently been found to promote cell survival, inhibiting the induction of apoptosis. In most cells other than B lymphocytes, NF-kappaB/Rel is inactive, sequestered in the cytoplasm. For example, nuclear extracts from two human untransformed breast epithelial cell lines expressed only very low levels of NF kappaB. Unexpectedly, nuclear extracts from two human breast tumor cell lines displayed significant levels of NF-kappaB/Rel. Direct inhibition of this NF kappaB/ Rel activity in breast cancer cells induced apoptosis. High levels of NF kappaB/Rel binding were also observed in carcinogen-induced primary rat mammary tumors, whereas only expectedly low levels were seen in normal rat mammary glands. Furthermore, multiple human breast cancer specimens contained significant levels of nuclear NF-kappaB/Rel subunits. Thus, aberrant nuclear expression of NF kappaB/Rel is associated with breast cancer. Given the role of NF-kappaB/Rel factors in cell survival, this aberrant activity may play a role in tumor progression, and represents a possible therapeutic target in the treatment of these tumors. PMID- 9399941 TI - Constitutive nuclear factor-kappaB-RelA activation is required for proliferation and survival of Hodgkin's disease tumor cells. AB - The pathogenesis and etiology of Hodgkin's disease, a common human malignant lymphoma, is still unresolved. As a unique characteristic, we have identified constitutive activation of the transcription factor nuclear factor (NF)-kappaB p50-RelA in Hodgkin/Reed-Sternberg (H/RS) cells, which discriminates these neoplastic cells from most cell types studied to date. In contrast to other lymphoid and nonlymphoid cell lines tested, proliferation of H/RS cells depended on activated NF-kappaB. Furthermore, constitutive NF-kappaB p50-RelA prevented Hodgkin's lymphoma cells from undergoing apoptosis under stress conditions. Consistent with this dual function, Hodgkin's lymphoma cells depleted of constitutive nuclear NF-kappaB revealed strongly impaired tumor growth in severe combined immunodeficient mice. Our findings identify NF-kappaB as an important component for understanding the pathogenesis of Hodgkin's disease and for developing new therapeutic strategies against it. PMID- 9399942 TI - Preferential localization of systemically administered radiolabeled interleukin 1alpha in experimental inflammation in mice by binding to the type II receptor. AB - Previously, we have shown that systemically administered radiolabeled interleukin 1alpha (IL-1alpha) accumulates preferentially in inflammatory foci in mice. Since inflammation is characterized by influx of leukocytes, which represent IL-1 receptor (IL-1R) positive cells, radiolabeled IL-1 may specifically localize in inflammation by binding to its receptors on infiltrated leukocytes. This hypothesis was tested in a series of studies in mice with acute focal inflammations. Evidence for specific IL-1-IL-1R interaction in induced inflammation was found: microscopic autoradiography revealed that 125I-IL-1alpha localized at the site of inflammatory cells with time; 125I-myoglobin, a similar sized protein with no known interactions in vivo, was not retained in the inflammation. Furthermore, the uptake 125I-IL-1alpha in inflammatory tissue was significantly lower in neutropenic mice than in immunocompetent mice (0.05+/ 0.004 vs. 0.65+/-0.06% ID/g at 48 h after injection, P < 0.0007). Moreover, the uptake of 125I-IL-1alpha at the inflammatory site could be blocked with the anti IL-1R type II antibody 4E2. At 48 h after injection, the uptake with and without blocking the type II IL-1R was 0.13+/-0.01 and 0. 65+/-0.05% ID/g, respectively (P < 0.0001). These in vivo studies provide evidence that systemically administered radiolabeled IL-1alpha localizes in inflammatory tissue by specific receptor binding, predominantly by binding to the type II IL-1R. PMID- 9399943 TI - The calcimimetic compound NPS R-568 suppresses parathyroid cell proliferation in rats with renal insufficiency. Control of parathyroid cell growth via a calcium receptor. AB - Parathyroid (PT) cell hyperplasia is a common consequence of chronic renal insufficiency (CRI). NPS R-568 is a phenylalkylamine compound that acts as an agonist (calcimimetic) at the cell surface calcium receptor (CaR). To test the hypothesis that the CaR plays a role in PT hyperplasia in CRI, we tested the effect of NPS R-568 on PT cell proliferation in rats with renal insufficiency. Rats were subjected to 5/6 nephrectomy and then infused intraperitoneally with 5 bromodeoxyuridine (BrdU) to label S-phase cells. Two groups of nephrectomized rats received NPS R-568 by gavage twice daily for 4 d (1.5 and 15 mg/kg body wt). On day 5, the number of BrdU-positive PT cells of vehicle-treated nephrectomized rats was 2.6-fold greater than that of the sham-operated control. Low and high doses of NPS R-568 reduced the number of BrdU-positive PT cells by 20 and 50%, respectively. No changes in staining, however, were observed in ileal epithelial cells (CaR-negative) or in thyroidal C-cells (CaR-positive). Furthermore, the effect of NPS R-568 could not be explained by changes in serum 1,25(OH)2D3 or phosphorus. These results indicate that NPS R-568 suppresses PT cell proliferation in rats with renal insufficiency, and lend support to the linkage between the CaR and PT hyperplasia in CRI. PMID- 9399944 TI - Role of nitric oxide in experimental obliterative bronchiolitis (chronic rejection) in the rat. AB - The role of nitric oxide in obliterative bronchiolitis development, i.e., chronic rejection, was investigated in the heterotopic rat tracheal allograft model. An increase in the intragraft inducible nitric oxide synthase (iNOS) mRNA and mononuclear inflammatory cell iNOS immunoreactivity was demonstrated during progressive loss of respiratory epithelium and airway occlusion in nontreated allografts compared to syngeneic grafts. In nontreated allografts, however, intragraft nitric oxide production was decreased, most likely because of loss of iNOS epithelial expression. Treatment with aminoguanidine, a preferential inhibitor of inducible nitric oxide synthase, was associated with enhanced proliferation of alpha-smooth muscle actin immunoreactive cells and the intensity of obliterative bronchiolitis early after transplantation. Aminoguanidine treatment did not affect iNOS mRNA synthesis or intragraft nitric oxide production, but decreased iNOS immunoreactivity in smooth muscle cells. Treatment with L-arginine, a precursor of nitric oxide, significantly reduced obliterative changes. L-arginine supplementation enhanced intragraft iNOS mRNA synthesis and iNOS immunoreactivity in capillary endothelial and smooth muscle cells as well as intragraft nitric oxide production. Immunohistochemical analysis of allografts showed that neither iNOS inhibition nor supplementation of the nitric oxide pathway affected the number of graft-infiltrating CD4+ and CD8+ T cells, ED1+ and ED3+ macrophages, immune activation with expression of IL-2R or MHC class II, or production of macrophage or Th1 cytokines. In contrast, L-arginine treatment was associated with increased staining for Th2 cytokines IL-4 and IL-10. In conclusion, this study demonstrates that nitric oxide has a protective role in obliterative bronchiolitis development in this model, and suggests that nitric oxide either directly or indirectly inhibits smooth muscle cell proliferation and modulates immune response towards Th2 cytokines. PMID- 9399945 TI - Immunohistochemical colocalization of glycoxidation products and lipid peroxidation products in diabetic renal glomerular lesions. Implication for glycoxidative stress in the pathogenesis of diabetic nephropathy. AB - Advanced glycation end products (AGEs) include a variety of protein adducts whose accumulation alters the structure and function of tissue proteins and stimulates cellular responses. They have been implicated in tissue damage associated with diabetic complications. To assess the possible link between AGE accumulation and the development of diabetic nephropathy (DN), we have examined the immunohistochemical localization of various AGE structures postulated to date, i.e., pentosidine, Nepsilon-(carboxymethyl)lysine (CML), and pyrraline, in diabetic and control kidneys. CML and pentosidine accumulate in the expanded mesangial matrix and thickened glomerular capillary walls of early DN and in nodular lesions and arterial walls of advanced DN, but were absent in control kidneys. By contrast, pyrraline was not found within diabetic glomeruli but was detected in the interstitial connective tissue of both normal and diabetic kidneys. Although the distribution of pyrraline was topographically identical to type III collagen, distribution of pentosidine and CML was not specific for collagen type, suggesting that difference in matrix protein composition per se could not explain heterogeneous AGE localization. Since oxidation is linked closely to the formation of pentosidine and CML, we also immunostained malondialdehyde (MDA), a lipid peroxidation product whose formation is accelerated by oxidative stress, assuming that local oxidative stress may serve as a mechanism of pentosidine and CML accumulation. Consistent with our assumption, diabetic nodular lesions were stained positive for MDA. These findings show that AGE localization in DN varies according to AGE structure, and suggest that the colocalization of markers of glycoxidation (pentosidine and CML) with a marker of lipid peroxidation reflects a local oxidative stress in association with the pathogenesis of diabetic glomerular lesions. Thus, glycoxidation markers may serve as useful biomarkers of oxidative damage in DN. PMID- 9399946 TI - Human beta-2 adrenoceptor gene polymorphisms are highly frequent in obesity and associate with altered adipocyte beta-2 adrenoceptor function. AB - Catecholamines play a central role in the regulation of energy expenditure, in part by stimulating lipid mobilization through lipolysis in fat cells. The beta-2 adrenoceptor (BAR-2) is a major lipolytic receptor in human fat cells. To determine whether known polymorphisms in codons 16, 27, and 164 of this receptor play a role in obesity and subcutaneous adipocyte BAR-2 lipolytic function, we investigated a group of 140 women with a large variation in body fat mass. Only the polymorphisms in codons 16 and 27 were common in the study population. The Gln27Glu polymorphism was markedly associated with obesity with a relative risk for obesity of approximately 7 and an odds ratio of approximately 10. Homozygotes for Glu27 had an average fat mass excess of 20 kg and approximately 50% larger fat cells than controls. However, no significant association with changes in BAR 2 function was observed. The Arg16Gly polymorphism was associated with altered BAR-2 function with Gly16 carriers showing a fivefold increased agonist sensitivity and without any change in BAR-2 expression. However, it was not significantly linked with obesity. These findings suggest that genetic variability in the human BAR-2 gene could be of major importance for obesity, energy expenditure, and lipolytic BAR-2 function in adipose tissue, at least in women. PMID- 9399947 TI - Immunization against the agent of human granulocytic ehrlichiosis in a murine model. AB - The agent of human granulocytic ehrlichiosis (HGE) is a newly recognized tick borne pathogen that resides within polymorphonuclear leukocytes. C3H/HeN mice can become infected with the agent of HGE (designated aoHGE) by syringe inoculation or tick-borne infection and develop transient neutropenia. They thereby partially mimic human disease and provide a model in which to study immunity to this microorganism. Mice vaccinated with lysates of purified aoHGE, or administered aoHGE antisera, were partially protected from both syringe- and tick-transmitted challenge with aoHGE. These data suggest that antibodies are sufficient to provide substantial, but not complete, immunity against aoHGE. PMID- 9399948 TI - An increased prevalence of Epstein-Barr virus infection in young patients suggests a possible etiology for systemic lupus erythematosus. AB - An unknown environmental agent has been suspected to induce systemic lupus erythematosus (lupus) in man. Prompted by our recent immunochemical findings, we sought evidence for an association between Epstein-Barr virus infection and lupus. Because the vast majority of adults have been infected with Epstein-Barr virus, we chose to study children and young adults. Virtually all (116 of 117, or 99%) of these young patients had seroconverted against Epstein-Barr virus, as compared with only 70% (107 of 153) of their controls (odds ratio 49.9, 95% confidence interval 9.3-1025, P < 0. 00000000001). The difference in the rate of Epstein-Barr virus seroconversion could not be explained by serum IgG level or by cross-reacting anti-Sm/nRNP autoantibodies. No similar difference was found in the seroconversion rates against four other herpes viruses. An assay for Epstein Barr viral DNA in peripheral blood lymphocytes established Epstein-Barr virus infection in the peripheral blood of all 32 of the lupus patients tested, while only 23 of the 32 matched controls were infected (odds ratio > 10, 95% confidence interval 2.53-infinity, P < 0.002). When considered with other evidence supporting a relationship between Epstein-Barr virus and lupus, these data are consistent with, but do not in themselves establish, Epstein-Barr virus infection as an etiologic factor in lupus. PMID- 9399950 TI - An interleukin-2 receptor gamma chain mutation with normal thymus morphology. AB - One of the most common human immunodeficiencies is an X-linked condition arising from mutations of the gamma subunit of the interleukin-2 receptor (IL-2Rgamma). The IL-2Rgamma protein is one chain of the heterotrimeric (alpha, beta, gamma) IL 2 receptor, but also participates in the formation of the IL-4, 7, 9, and 15 receptor complexes. The diagnosis of X-linked SCID is usually relatively simple due to the distinctive immunological presentation; IL-2Rgamma-deficient patients typically lacking mature T lymphocytes (T-B+). However, it is becoming clear that this merely represents one extreme of a potential range of clinical presentations. We describe here a novel mutation of the human IL-2Rgamma chain (R222C) resulting in an unusual immunological phenotype. Although clinically immunodeficient, this patient has normal numbers of peripheral T and B cells, responds normally to mitogenic stimuli, and unusually, has a normal thymus gland. This IL-2Rgamma mutation is distinctive in that the protein is sufficiently stable to be expressed at the cell surface. While the T cell receptor repertoire appears complete, suggesting normal T cell differentiation occurs, patient T cells demonstrate a reduced ability to bind IL-2 and this appears sufficient to cause a deficiency in their ability to participate in antigenic responses. Early clinical recognition of this phenotype is critical as a delay in diagnosis may result in a fatal infection. PMID- 9399949 TI - Prevention of experimental myasthenia gravis by nasal administration of synthetic acetylcholine receptor T epitope sequences. AB - T cell tolerization prevents and improves T cell-mediated experimental autoimmune diseases. We investigated here whether similar approaches could be used for antibody (Ab)-mediated autoimmune diseases. Myasthenia gravis, caused by IgG Ab against muscle acetylcholine receptor (AChR), is perhaps the best characterized of them. We used an animal model, experimental myasthenia gravis induced in C57Bl/6 mice by immunization with Torpedo acetylcholine receptor (TAChR), to demonstrate that nasal administration of synthetic sequences of the TAChR alpha subunit- forming epitopes recognized by anti-TAChR CD4+ T helper cells (residues alpha150-169, alpha181-200, and alpha360-378), given before and during immunization with TAChR, causes decreased CD4+ responsiveness to those epitopes and to TAChR, reduced synthesis of anti-TAChR Ab, and prevented experimental myasthenia gravis. These effects were not induced by nasal administration of synthetic epitopes of diphtheria toxin. Secretion of IL-2, IL-4, and IL-10 by spleen T cells from TAChR immunized mice, in response to challenge with TAChR in vitro, indicated that in sham-tolerized mice only Th1 cells responded to TAChR, while peptide-treated mice had also an AChR-specific Th2 response. The TAChR peptide treatment induced also in vitro anergy to the TAChR of the spleen T cells, which was reversed by IL-2. PMID- 9399951 TI - Overexpression of Rab3D enhances regulated amylase secretion from pancreatic acini of transgenic mice. AB - Rab3D, a member of the ras-related GTP-binding protein Rab family, is localized to secretory granules of various exocrine tissues such as acinar cells of the pancreas, chief cells of the stomach, and parotid and lacrimal secretory cells. To elucidate the function of Rab3D in exocytosis, we have generated transgenic mice that over-express Rab3D specifically in pancreatic acinar cells. Hemagglutinin-tagged Rab3D was localized to zymogen granules by immunohistochemistry, and was shown to be present on zymogen granule membranes by Western blotting; both results are similar to previous studies of endogenous Rab3D. Secretion measurements in isolated acinar preparations showed that overexpression of Rab3D enhanced amylase release. Amylase secretion from intact acini of transgenic mice 5 min after 10 pM cholecystokinin octapeptide (CCK) stimulation was enhanced by 160% of control. In streptolysin-O-permeabilized acini of transgenic mice, amylase secretion induced by 100 microM GTP-gamma-S was enhanced by 150%, and 10 microM Ca2+-stimulated amylase secretion was augmented by 206% of that of the control. To further elucidate Rab3D involvement in stimulus-secretion coupling, we examined the effect of CCK on the rate of GTP binding to Rab3D. Stimulation of permeabilized acini with 10 pM CCK increased the incorporation of radiolabeled GTP into HA-tagged Rab3D. These results indicate that overexpression of Rab3D enhances secretagogue-stimulated amylase secretion through both calcium and GTP pathways. We conclude that Rab3D protein on zymogen granules plays a stimulatory role in regulated amylase secretion from pancreatic acini. PMID- 9399952 TI - Myocardial ischemia induces differential regulation of KATP channel gene expression in rat hearts. AB - The cardiac ATP-sensitive potassium (KATP) channel is thought to be a complex composed of an inward rectifier potassium channel (Kir6.1 and/or Kir6.2) subunit and the sulfonylurea receptor (SUR2). This channel is activated during myocardial ischemia and protects the heart from ischemic injury. We examined the transcriptional expression of these genes in rats with myocardial ischemia. 60 min of myocardial regional ischemia followed by 24-72 h, but not 3-6 h, of reperfusion specifically upregulated Kir6.1 mRNA not only in the ischemic (approximately 2.7-3.1-fold) but also in the nonischemic (approximately 2.0-2.6 fold) region of the left ventricle. 24 h of continuous ischemia without reperfusion also induced an increase in Kir6.1 mRNA in both regions, whereas 15 30 min of ischemia followed by 24 h of reperfusion did not induce such expression. In contrast, mRNAs for Kir6.2 and SUR2 remained unchanged under these ischemic procedures. Western blotting demonstrated similar increases in the Kir6.1 protein level both in the ischemic (2.4-fold) and the nonischemic (2.2 fold) region of rat hearts subjected to 60 min of ischemia followed by 24 h of reperfusion. Thus, prolonged myocardial ischemia rather than reperfusion induces delayed and differential regulation of cardiac KATP channel gene expression. PMID- 9399953 TI - Lung disease in mice with cystic fibrosis. AB - The leading cause of mortality and morbidity in humans with cystic fibrosis is lung disease. Advances in our understanding of the pathogenesis of the lung disease of cystic fibrosis, as well as development of innovative therapeutic interventions, have been compromised by the lack of a natural animal model. The utility of the CFTR-knockout mouse in studying the pathogenesis of cystic fibrosis has been limited because of their failure, despite the presence of severe intestinal disease, to develop lung disease. Herein, we describe the phenotype of an inbred congenic strain of CFTR-knockout mouse that develops spontaneous and progressive lung disease of early onset. The major features of the lung disease include failure of effective mucociliary transport, postbronchiolar over inflation of alveoli and parenchymal interstitial thickening, with evidence of fibrosis and inflammatory cell recruitment. We speculate that the basis for development of lung disease in the congenic CFTR knockout mice is their observed lack of a non-CFTR chloride channel normally found in CFTR-knockout mice of mixed genetic background. PMID- 9399954 TI - Microtubule-associated protein 1 light chain 3 is a fibronectin mRNA-binding protein linked to mRNA translation in lamb vascular smooth muscle cells. AB - Intimal cushions form in the fetal ductus arteriosus by fibronectin-dependent smooth muscle cell migration which is associated with greater efficiency of fibronectin mRNA translation. We investigated whether the AU-rich element (ARE), UUAUUUAU, in the 3'-untranslated region (3'UTR) of fibronectin mRNA is involved in this mechanism by transfecting smooth muscle cells with plasmids containing the chloramphenicol acetyltransferase coding region with its 3'UTR replaced by fibronectin 3'UTR bearing intact or mutated ARE. More efficient translation of fusion mRNA with intact versus mutated ARE was observed. This effect was amplified in ductus (10.9-fold) compared with nonmigratory, lower fibronectin producing aorta cells (6.5-fold). Ductus cells transfected with wild-type but not ARE-mutated plasmid reverted to the stellate phenotype of aorta cells associated with reduced fibronectin production. This suggested that plasmid ARE sequesters RNA-binding factors, thereby reducing endogenous fibronectin mRNA translation. We next purified a 15-kD fibronectin ARE-dependent RNA-binding protein and identified it as microtubule-associated protein 1 light chain 3 (LC3). LC3 is present in greater amounts in ductus compared with aorta cells, and overexpression of LC3 in aortic cells by transfection enhances fibronectin mRNA translation to levels observed in ductus cells. PMID- 9399955 TI - Blockade of CD49d (alpha4 integrin) on intrapulmonary but not circulating leukocytes inhibits airway inflammation and hyperresponsiveness in a mouse model of asthma. AB - Immunized mice after inhalation of specific antigen have the following characteristic features of human asthma: airway eosinophilia, mucus and Th2 cytokine release, and hyperresponsiveness to methacholine. A model of late-phase allergic pulmonary inflammation in ovalbumin-sensitized mice was used to address the role of the alpha4 integrin (CD49d) in mediating the airway inflammation and hyperresponsiveness. Local, intrapulmonary blockade of CD49d by intranasal administration of CD49d mAb inhibited all signs of lung inflammation, IL-4 and IL 5 release, and hyperresponsiveness to methacholine. In contrast, CD49d blockade on circulating leukocytes by intraperitoneal CD49d mAb treatment only prevented the airway eosinophilia. In this asthma model, a CD49d-positive intrapulmonary leukocyte distinct from the eosinophil is the key effector cell of allergen induced pulmonary inflammation and hyperresponsiveness. PMID- 9399956 TI - Characterization of the AD7C-NTP cDNA expression in Alzheimer's disease and measurement of a 41-kD protein in cerebrospinal fluid. AB - We have isolated a novel Alu sequence-containing cDNA, designated AD7c-NTP, that is expressed in neurons, and overexpressed in brains with Alzheimer's disease (AD). The 1,442-nucleotide AD7c-NTP cDNA encodes an approximately 41-kD protein. Expression of AD7c-NTP was confirmed by nucleic acid sequencing of reverse transcriptase PCR products isolated from brain. AD7c-NTP cDNA probes hybridized with 1. 4 kB mRNA transcripts by Northern blot analysis, and monoclonal antibodies generated with the recombinant protein were immunoreactive with approximately 41-45-kD and approximately 18-21-kD molecules by Western blot analysis. In situ hybridization and immunostaining studies localized AD7c-NTP gene expression in neurons. Using a quantitative enzyme-linked sandwich immunoassay (Ghanbari, K., I. Beheshti, and H. Ghanbari, manuscript submitted for publication) constructed with antibodies to the recombinant protein, AD7c-NTP levels were measured under code in 323 clinical and postmortem cerebrospinal fluid (CSF) samples from AD, age-matched control, Parkinson's disease, and neurological disease control patients. The molecular mass of the AD7c-NTP detected in CSF was approximately 41 kD. In postmortem CSF, the mean concentration of AD7c-NTP in cases of definite AD (9.2+/-8.2 ng/ml) was higher than in the aged control group (1.6+/-0.9; P < 0.0001). In CSF samples from individuals with early possible or probable AD, the mean concentration of AD7c NTP (4.6+/-3.4) was also elevated relative to the levels in CSF from age-matched (1.2+/-0.7) and neurological disease (1.0+/-0.9) controls, and ambulatory patients with Parkinson's disease (1.8+/-1.1) (all P < 0.001). CSF levels of AD7c NTP were correlated with Blessed dementia scale scores (r = 0. 66; P = 0.0001) rather than age (r = -0.06; P > 0.1). In vitro studies demonstrated that overexpression of AD7c-NTP in transfected neuronal cells promotes neuritic sprouting and cell death, the two principal neuroanatomical lesions correlated with dementia in AD. The results suggest that abnormal AD7c-NTP expression is associated with AD neurodegeneration, and during the early stages of disease, CSF levels correlate with the severity of dementia. PMID- 9399957 TI - Leptin selectively decreases visceral adiposity and enhances insulin action. AB - Intraabdominal adiposity and insulin resistance are risk factors for diabetes mellitus, dyslipidemia, arteriosclerosis, and mortality. Leptin, a fat-derived protein encoded by the ob gene, has been postulated to be a sensor of energy storage in adipose tissue capable of mediating a feedback signal to sites involved in the regulation of energy homeostasis. Here, we provide evidence for specific effects of leptin on fat distribution and in vivo insulin action. Leptin (LEP) or vehicle (CON) was administered by osmotic minipumps for 8 d to pair-fed adult rats. During the 8 d of the study, body weight and total fat mass decreased similarly in LEP and in CON. However, while moderate calorie restriction (CON) resulted in similar decreases in whole body (by 20%) and visceral (by 21%) fat, leptin administration led to a specific and marked decrease (by 62%) in visceral adiposity. During physiologic hyperinsulinemia (insulin clamp), leptin markedly enhanced insulin action on both inhibition of hepatic glucose production and stimulation of glucose uptake. Finally, leptin exerted complex effects on the hepatic gene expression of key metabolic enzymes and on the intrahepatic partitioning of metabolic fluxes, which are likely to represent a defense against excessive storage of energy in adipose depots. These studies demonstrate novel actions of circulating leptin in the regulation of fat distribution, insulin action, and hepatic gene expression and suggest that it may play a role in the pathophysiology of abdominal obesity and insulin resistance. PMID- 9399958 TI - The antifungal antibiotic, clotrimazole, inhibits chloride secretion by human intestinal T84 cells via blockade of distinct basolateral K+ conductances. Demonstration of efficacy in intact rabbit colon and in an in vivo mouse model of cholera. AB - The antifungal antibiotic clotrimazole (CLT) blocks directly and with high potency the Ca2+-activated K+ channels of human erythrocytes, erythroleukemia cells, and ferret vascular smooth muscle cells. We recently reported that CLT inhibits Cl- secretion in human intestinal T84 cells, likely by affecting K+ transport (Rufo, P.A., L. Jiang, S.J. Moe, C. Brugnara, S.L. Alper, and W.I. Lencer. 1996. J. Clin. Invest. 98:2066-2075). To determine if CLT had direct effects on K+ conductances in T84 cells, we selectively permeabilized apical membranes of confluent T84 cell monolayers using the ionophore amphotericin B. This technique permits direct measurement of basolateral K+ transport. We found that CLT and a stable des-imidazolyl derivative inhibited directly two pharmacologically distinct basolateral membrane K+conductances, but had no effect on apical membrane Cl- conductances. The effects of CLT on Cl- secretion were also examined in intact tissue. CLT inhibited forskolin-induced Cl- secretion in rabbit colonic mucosal sheets mounted in Ussing chambers by 91%. CLT also inhibited cholera toxin-induced intestinal Cl- secretion in intact mice by 94%. These data provide direct evidence that CLT blocks Cl- secretion in intestinal T84 cells by inhibition of basolateral K+ conductances, and show that CLT inhibits salt and water secretion from intact tissue in vitro and in vivo. The results further support the suggestion that CLT and its metabolites may show clinical efficacy in the treatment of secretory diarrheas of diverse etiologies. PMID- 9399959 TI - Indirect effect of insulin to suppress endogenous glucose production is dominant, even with hyperglucagonemia. AB - Suppression of endogenous glucose production (EGP) is one of insulin's primary metabolic effects and failure of this action is a major contributor to fasting hyperglycemia of type 2 diabetes mellitus. Classically, insulin was thought to suppress the liver directly, via hyperinsulinemia in the portal vein. Recently, however, we and others have demonstrated that at least part, and possibly most of insulin's action to suppress EGP is normally mediated via an extrahepatic (i.e., indirect) mechanism. We have suggested that this mechanism involves insulin suppression of adipocyte lipolysis, leading to lowered FFA and reduced EGP ("Single Gateway Hypothesis"). Previous studies of the indirect insulin effect from this laboratory were done under conditions of lowered portal glucagon. Because of the possibility that the direct (i.e., portal) effect of insulin may have been underestimated with hypoglucagonemia, these studies examined the relative importance of portal insulin, versus peripheral insulin (administered at one-half the dose to equalize peripheral insulin levels) at four rates of portal glucagon infusion: 0, 0.65 (under-), 1.5 (basal-), and 3.0 ng/kg per min (over replacement). Portal versus peripheral insulin suppressed steady-state EGP to the same extent (52%), confirming that the primary effect of insulin to suppress EGP is via the peripheral mechanism. This conclusion was maintained regardless of portal glucagonemia, although there was some evidence for an increase in the direct insulin effect at hyperglucagonemia. The indirect effect of insulin is the primary mechanism of steady-state EGP suppression under normal conditions. The direct effect increases with hyperglucagonemia; however, the indirect effect remains predominant even under those conditions. PMID- 9399961 TI - B lymphocytes from patients with chronic lymphocytic leukemia contain signal transducer and activator of transcription (STAT) 1 and STAT3 constitutively phosphorylated on serine residues. AB - To explore the pathogenesis of chronic lymphocytic leukemia (CLL), we examined whether phosphorylation of one or more signal transducer and activator of transcription (STAT) factors was abnormal in cells from CLL patients. No constitutive tyrosine phosphorylation was detected on any STAT in CLL cells. To assess the phosphorylation of serine residues of STAT1 and STAT3 in CLL cells, we raised antibodies that specifically recognize the form of STAT1 phosphorylated on ser-727 and the form of STAT3 phosphorylated on ser-727. We found that in 100% of patients with CLL (n = 32), STAT1 and STAT3 were constitutively phosphorylated on serine. This was in contrast to normal peripheral blood B lymphocytes or CD5+) B cells isolated from tonsils, in which this phosphorylation was absent. Serine phosphorylation of STAT1 and STAT3 was seen occasionally in other leukemias, but it was a universal finding only in CLL. The serine phosphorylation of these STATs was a continuous process, as incubation of CLL cells with the kinase inhibitor H7 led to the dephosphorylation of these serine residues. The STAT serine kinase in CLL cells has not been identified, and appears to be neither mitogen-activated protein kinase nor pp70(s6k). In summary, the constitutive serine phosphorylation of STAT1 and STAT3 is present in all CLL samples tested to date, although the physiologic significance of this modification remains to be determined. PMID- 9399960 TI - Nitric oxide production contributes to the angiogenic properties of vascular endothelial growth factor in human endothelial cells. AB - Vascular endothelial growth factor (VEGF) is a regulator of vasculogenesis and angiogenesis. To investigate the role of nitric oxide (NO) in VEGF-induced proliferation and in vitro angiogenesis, human umbilical vein endothelial cells (HUVEC) were used. VEGF stimulated the growth of HUVEC in an NO-dependent manner. In addition, VEGF promoted the NO-dependent formation of network-like structures in HUVEC cultured in three dimensional (3D) collagen gels. Exposure of cells to VEGF led to a concentration-dependent increase in cGMP levels, an indicator of NO production, that was inhibited by nitro-L-arginine methyl ester. VEGF-stimulated NO production required activation of tyrosine kinases and increases in intracellular calcium, since tyrosine kinase inhibitors and calcium chelators attenuated VEGF-induced NO release. Moreover, two chemically distinct phosphoinositide 3 kinase (PI-3K) inhibitors attenuated NO release after VEGF stimulation. In addition, HUVEC incubated with VEGF for 24 h showed an increase in the amount of endothelial NO synthase (eNOS) protein and the release of NO. In summary, both short- and long-term exposure of human EC to VEGF stimulates the release of biologically active NO. While long-term exposure increases eNOS protein levels, short-term stimulation with VEGF promotes NO release through mechanisms involving tyrosine and PI-3K kinases, suggesting that NO mediates aspects of VEGF signaling required for EC proliferation and organization in vitro. PMID- 9399962 TI - Activators of peroxisome proliferator-activated receptor gamma have depot specific effects on human preadipocyte differentiation. AB - Activation of peroxisome proliferator-activated receptor (PPAR) gamma, a nuclear receptor highly expressed in adipocytes, induces the differentiation of murine preadipocyte cell lines. Recently, thiazolidinediones (TZDs), a novel class of insulin-sensitizing compounds effective in the treatment of non-insulin-dependent diabetes mellitus (NIDDM) have been shown to bind to PPARgamma with high affinity. We have examined the effects of these compounds on the differentiation of human preadipocytes derived from subcutaneous (SC) and omental (Om) fat. Assessed by lipid accumulation, glycerol 3-phosphate dehydrogenase activity, and mRNA levels, subcultured preadipocytes isolated from either SC or Om depots did not differentiate in defined serum-free medium. Addition of TZDs (BRL49653 or troglitazone) or 15-deoxyDelta12,14prostaglandin J2 (a natural PPARgamma ligand) enhanced markedly the differentiation of preadipocytes from SC sites, assessed by all three criteria. The rank order of potency of these agents in inducing differentiation matched their ability to activate transcription via human PPARgamma. In contrast, preadipocytes from Om sites in the same individuals were refractory to TZDs, although PPARgamma was expressed at similar levels in both depots. The mechanism of this depot-specific TZD response is unknown. However, given the association between Om adiposity and NIDDM, the site-specific responsiveness of human preadipocytes to TZDs may be involved in the beneficial effects of these compounds on in vivo insulin sensitivity. PMID- 9399963 TI - Interferon-gamma and tumor necrosis factor-alpha specifically induce formation of cytomegalovirus-permissive monocyte-derived macrophages that are refractory to the antiviral activity of these cytokines. AB - Monocytes/macrophages are key cells in the pathogenesis of human cytomegalovirus (HCMV). Although HCMV infection in monocytes is restricted to early events of gene expression, productive infection has been demonstrated in differentiated macrophages in vitro. We examined the cellular and cytokine components that are essential for HCMV replication in Concanavalin A-stimulated monocyte-derived macrophages (MDM). By negative selection, depletion of CD8+ T lymphocytes, but not CD4+ T lymphocytes, CD19+ B cells, or CD56+ NK cells, resulted in a 60-70% reduction in the number of HCMV-infected MDM, and a 4 log decrease in virus production. Neutralization of IFN-gamma and TNF-alpha, but not IL-1, IL-2, or TGF beta, decreased production of virus by 4 logs and 2 logs, respectively. Subsequently, addition of recombinant IFN-gamma or TNF-alpha to purified monocyte cultures was sufficient to produce HCMV-permissive MDM. While IFN-gamma and TNF alpha possess antiviral properties, addition of these cytokines to permissive MDM cultures did not affect production of HCMV. Thus, rather than inhibiting replication of HCMV, IFN-gamma and TNF-alpha specifically induce differentiation of monocytes into HCMV-permissive MDM, which are resistant to the antiviral effects of these cytokines. PMID- 9399964 TI - Differential regulation of insulin receptor substrates-1 and -2 (IRS-1 and IRS-2) and phosphatidylinositol 3-kinase isoforms in liver and muscle of the obese diabetic (ob/ob) mouse. AB - Intracellular insulin signaling involves a series of alternative and complementary pathways created by the multiple substrates of the insulin receptor (IRS) and the various isoforms of SH2 domain signaling molecules that can interact with these substrates. In this study, we have evaluated the roles of IRS 1 and IRS-2 in signaling to the phosphatidylinositol (PI) 3-kinase pathway in the ob/ob mouse, a model of the insulin resistance of obesity and non-insulin dependent diabetes mellitus. We find that the levels of expression of both IRS-1 and IRS-2 are decreased approximately 50% in muscle, whereas in liver the decrease is significantly greater for IRS-2 (72%) than for IRS-1 (29%). This results in differential decreases in IRS-1 and IRS-2 phosphorylation, docking of the p85alpha regulatory subunit of PI 3-kinase, and activation of this enzyme in these two insulin target tissues. In ob/ob liver there is also a change in expression of the alternatively spliced isoforms of the regulatory subunits for PI 3-kinase that was detected at the protein and mRNA level. This resulted in a 45% decrease in the p85alpha form of PI 3-kinase, a ninefold increase in the AS53/p55alpha, and a twofold increase in p50alpha isoforms. Thus, there are multiple alterations in the early steps of insulin signaling in the ob/ob mouse, with differential regulation of IRS-1 and IRS-2, various PI 3-kinase regulatory isoforms, and a lack of compensation for the decrease in insulin signaling by any of the known alternative pathways at these levels. PMID- 9399965 TI - Following the fate of individual T cells throughout activation and clonal expansion. Signals from T cell receptor and CD28 differentially regulate the induction and duration of a proliferative response. AB - A detailed understanding of the effects of costimulatory signals on primary T cell expansion has been limited by experimental approaches that measure the bulk response of a cell population, without distinguishing responses of individual cells. Here, we have labeled live T cells in vitro with a stable, fluorescent dye that segregates equally between daughter cells upon cell division, allowing the proliferative history of any T cell present or generated during a response to be monitored over time. This system permits simultaneous evaluation of T cell surface markers, allowing concomitant assessment of cellular activation and quantitative determination of T cell receptor (TCR) occupancy on individual cells. Through this approach, we find that TCR engagement primarily regulates the frequency of T cells that enter the proliferative pool, but has relatively little effect on the number of times these cells will ultimately divide. In contrast, CD28-costimulation regulates both the frequency of responding cells (particularly at sub-maximal levels of TCR engagement), and more prominently, the number of mitotic events that responding cells undergo. When CD28-stimulation is blocked, provision of IL-2 restores the frequency of responding cells and the normal pattern of mitotic progression, indicating that the other CD28-induced genes are not required for this effect. An unexpected finding was that even at maximal levels of TCR engagement and CD28-mediated costimulation, only 50-60% of the original T cells in culture can be induced to divide. The nondividing cells are heterogeneous for naive versus memory markers, suggesting a more complex relationship between expression of memory markers and the ability to be recruited into the dividing pool. From these studies, we conclude that a stringent checkpoint regulates the participation of activated T cells in clonal expansion, with TCR and CD28 signals having both overlapping and differential effects on the induction and maintenance of T cell responses. PMID- 9399966 TI - Association between genetic polymorphisms of the beta2-adrenoceptor and response to albuterol in children with and without a history of wheezing. AB - The beta2-adrenergic receptor (beta2AR) agonists are the most widely used agents in the treatment of asthma, but the genetic determinants of responsiveness to these agents are unknown. Two polymorphic loci within the coding region of the beta2AR have been recently described at amino acids 16 and 27. It has been reported that glycine at codon 16 (Gly-16) is associated with increased agonist promoted downregulation of the beta2AR as compared with arginine-16 (Arg-16). The form of the receptor with glutamic acid at codon 27 (Glu-27), on the other hand, has been shown to be resistant to downregulation when compared with glutamine-27 (Gln-27), but only when coexpressed with Arg-16. To assess if different genotypes of these two polymorphisms would show differential responses to inhaled beta2AR agonists, we genotyped 269 children who were participants in a longitudinal study of asthma. Spirometry was performed before and after administration of 180 microg of albuterol, and a positive response was considered an increase of >15.3% predicted FEV1. There was marked linkage disequilibrium between the two polymorphisms, with 97.8% of all chromosomes that carried Arg-16 also carrying Gln-27. When compared to homozygotes for Gly-16, homozygotes for Arg-16 were 5.3 times (95% confidence interval 1.6-17.7) and heterozygotes for beta2AR-16 were 2.3 times (1.3-4.2) more likely to respond to albuterol, respectively. Similar trends were observed for asthmatic and nonasthmatic children, and results were independent of baseline lung function, ethnic origin, and previous use of antiasthma medication. No association was found between the beta2AR-27 polymorphism and response to albuterol. These results may explain some of the variability in response to therapeutic doses of albuterol in children. PMID- 9399967 TI - Anabaseine is a potent agonist on muscle and neuronal alpha-bungarotoxin sensitive nicotinic receptors. AB - We assessed the pharmacological activity of anabaseine, a toxin found in certain animal venoms, relative to nicotine and anabasine on a variety of vertebrate nicotinic receptors, using cultured cells, the Xenopus oocyte expression system, contractility assays with skeletal and smooth muscle strips containing nicotinic receptors and in vivo rat prostration assay involving direct injection into the lateral ventricle of the brain. Anabaseine stimulated every subtype of nicotinic receptor that was tested. It was the most potent frog skeletal muscle nicotinic receptor agonist. At higher concentrations it also blocked the BC3H1 (adult mouse) muscle type receptor ion channel. The affinities of the three nicotinoid compounds for rat brain membrane alpha-bungarotoxin binding sites and their potencies for stimulating Xenopus oocyte homomeric alpha7 receptors, expressed in terms of their active monocation concentrations, displayed the same rank order, anabaseine>anabasine> nicotine. Although the maximum currents generated by anabaseine and anabasine at alpha7 receptors were equivalent to that of acetylcholine, the maximum response to nicotine was only about 65% of the acetylcholine response. At alpha4-beta2 receptors the affinities and apparent efficacies of anabaseine and anabasine were much less than that of nicotine. Anabaseine, nicotine and anabasine were nearly equipotent on sympathetic (PC12) receptors, although parasympathetic (myenteric plexus) receptors were much more sensitive to anabaseine and nicotine but less sensitive to anabasine. These differences suggest that there may be different subunit combinations in these two autonomic nicotinic receptors. The preferential interactions of anabaseine, anabasine and nicotine with different receptor subtypes provides molecular clues that should be helpful in the design of selective nicotinic agonists. PMID- 9399968 TI - Nifedipine, an L-type calcium channel blocker, restores the hypnotic response in rats made tolerant to the alpha-2 adrenergic agonist dexmedetomidine. AB - Rats were made tolerant to the hypnotic effects of the alpha-2 adrenergic agonist dexmedetomidine by a 7- or 14-day continuous systemic administration of the same, and the ability of nifedipine to reverse dexmedetomidine tolerance was assessed. Acute administration of nifedipine (10 mg/kg i.p.) restored the hypnotic response to dexmedetomidine in the alpha-2 tolerant rats. Concurrent administration of nifedipine during induction of tolerance, either partially (continuous administration 10 mg/kg/day delivered by minipumps) or completely (twice daily injections, 20 mg/kg s.c.) restored hypnotic responsiveness to control levels. Induction of tolerance reduced the affinity of [3H]PN200-110 for the L-type calcium channel. Chronically administered nifedipine treatment (20 mg/kg s.c. twice daily), at doses that partially restored the behavioral response to normal, did not change ligand binding affinity of [3H]PN200-110. An increase in Bmax for [3H]PN200-110 was noted in the dexmedetomidine tolerant state which did not change with chronic nifedipine. In naive rats, the phosphodiesterase inhibitor rolipram (275 microg/kg i.p.), mimicked the state of tolerance, as it resulted in a decreased hypnotic response to dexmedetomidine. Nifedipine (10 mg/kg i.p.) also reversed the rolipram-induced attenuation of the hypnotic response to dexmedetomidine. These data implicate a role for the L-type calcium channel in the mechanism of the hypnotic response in alpha-2 tolerant rats and suggest the involvement of the cAMP pathway. PMID- 9399969 TI - Comparison of effect of mosapride citrate and existing 5-HT4 receptor agonists on gastrointestinal motility in vivo and in vitro. AB - Mosapride citrate is a new gastroprokinetic agent that enhances the upper GI motility by stimulating 5-hydroxytryptamine4 (5-HT4) receptors. The purpose of this study was to compare the effects of mosapride and the existing 5-HT4 receptor agonists on GI motility in conscious dogs and on various 5-HT4 receptor mediated responses in vitro. In conscious dogs with force transducers implanted, mosapride (0.3-3 mg/kg i.v.) stimulated the antral motility without affecting the colonic motility. However, cisapride, zacopride and BIMU 8 (0. 1-1 mg/kg i.v.) stimulated both antral and colonic motility. The enhanced GI motility induced by mosapride or cisapride was antagonized by pretreatment with GR113808 (1 mg/kg bolus i.v., thereafter 1 mg/kg/hr infusion), a selective 5-HT4 receptor antagonist. In the receptor binding studies, mosapride inhibited [3H]-GR113808 binding to 5-HT4 receptor sites of guinea pig striatum with an IC50 value of 113 nM. In addition, mosapride caused relaxation of the carbachol-precontracted rat esophagus, enhanced the electrically evoked contractions of guinea pig ileum and evoked the contractions of guinea pig distal colon with EC50 values of 208, 73, and 3029 nM, respectively; this indicates that mosapride has a low affinity for colon than for the rest of the GI tract. In contrast, cisapride, zacopride or BIMU 8 had similar potencies in all preparations examined. In conclusion, these studies indicate that mosapride selectively stimulates upper GI motility in vivo and in vitro. These results also suggest heterogeneity of 5-HT4 receptors in the GI tract. PMID- 9399970 TI - Discriminative stimulus effects of the mixed-opioid agonist/antagonist dezocine: cross-substitution by mu and delta opioid agonists. AB - The purpose of this investigation was to evaluate the discriminative stimulus effects of the mixed-opioid agonist/antagonist dezocine. In pigeons trained to discriminate 1.7 mg/kg dezocine from saline, a series of opioids with activity at the mu opioid receptor substituted completely for the dezocine stimulus with a rank order of potency similar to that obtained in other assays sensitive to the effects of mu agonists (i.e., fentanyl >[-]-cyclazocine >buprenorphine = butorphanol >l-methadone >nalbuphine >[-]-metazocine >morphine). (-)-N allylnormetazocine and (+)-propoxyphene substituted partially for the dezocine stimulus, an effect obtained even when tested up to doses that suppressed responding. Naloxone (0.1 - 10 mg/kg) antagonized the stimulus effects of dezocine, (+)-propoxyphene and fentanyl in a dose-related manner, whereas doses of naloxone that antagonized fentanyl's rate-decreasing effects failed to antagonize the rate-decreasing effects of dezocine and (+)-propoxyphene. A 10 mg/kg dose of the mu-selective, noncompetitive antagonist beta-funaltrexamine was more effective against the stimulus effects of dezocine and nalbuphine than against morphine and fentanyl. As was observed with naloxone, beta-funaltrexamine failed to antagonize dezocine's rate-decreasing effects. The delta agonists BW373U86 and SNC80 substituted partially for the dezocine stimulus, and these effects were reversed by doses of the delta-selective antagonist naltrindole (0.1 and 1.0 mg/kg) that had no effect on the dezocine stimulus. Naltrindole also antagonized the rate-decreasing effects produced by BW373U86 and SNC80, but not those of dezocine. The kappa agonists bremazocine, spiradoline, U50,488 and U69,593 failed to substitute for the dezocine stimulus. The kappa-selective antagonist norbinaltorphimine (1.0 mg/kg) failed to antagonize dezocine's stimulus or rate-decreasing effects. The present findings indicate that dezocine shares similar stimulus effects with both mu and delta agonists, its stimulus effects are reversed by mu-selective antagonists, and its rate-decreasing effects are not mediated by activity at mu, kappa or delta opioid receptors. PMID- 9399971 TI - Selective brain to blood efflux transport of para-aminohippuric acid across the blood-brain barrier: in vivo evidence by use of the brain efflux index method. AB - Efflux transport of para-aminohippuric acid (PAH) across the blood-brain barrier (BBB) has been demonstrated by use of the brain efflux index (BEI) method. PAH was eliminated from the ipsilateral cerebrum extensively with an apparent efflux rate constant of 0.0587 (min-1) after microinjection into a cerebral cortex region termed Par2. This efflux transport showed a saturation with the Michaelis constant of approximately 400 microM. No more than 3% dose of PAH and carboxyl inulin, used as a reference compound showing limited permeability at the BBB, were found in the contralateral cerebrum, cerebellum or cerebrospinal fluid up to 20 min after administration. Under saturated conditions for carrier-mediated efflux of PAH via the blood-cerebrospinal fluid barrier, the BEI value of PAH did not change significantly, which suggested that blood-cerebrospinal fluid barrier was not responsible for the elimination of PAH from the brain after microinjection. No significant metabolism of PAH was demonstrated in the brain for at least 20 min after microinjection, and most of the radioactivity in the ipsilateral and contralateral carotid veins was as the intact form. With the distribution volume of PAH, 0.800 ml/g brain, obtained from the brain slice uptake experiment, the apparent efflux clearance was calculated as 46.9 microl/min/g brain. In addition, the influx clearance of PAH across the BBB determined by the in vivo brain uptake index method was much smaller than the efflux clearance, which demonstrates that BBB transports PAH selectively from the brain to the circulating blood. PMID- 9399972 TI - Effects of tachykinins on rapidly adapting pulmonary stretch receptors and total lung resistance in anesthetized, artificially ventilated rabbits. AB - In anesthetized, artificially ventilated rabbits not treated with thiorphan (2 mg/kg), a neutral endopeptidase (NEP) inhibitor, substance P (SP) and neurokinin A (NKA) in doses from 0.2 to 2.7 microg/kg produced dose-related increases in rapidly adapting pulmonary stretch receptor (RAR) activity without any significant changes in total lung resistance (RL), whereas neurokinin B (NKB) at the same concentrations did not significantly alter either RAR activity or RL. In comparison with the excitatory responses of RAR activity to SP and NKA, the magnitudes of increased receptor activity evoked SP were significantly larger than those after NKA administration. The rank order of tachykinins for RAR stimulus potency was SP > NKA > KB. Pretreatment with thiorphan potentiated the increases of RAR activity and RL induced by SP but had no effect on the RAR and RL responses to NKA and NKB. Subsequent administration of L 659, 877 (a selective NK2 receptor antagonist, 2. 3 and 7.6 microg/kg) that dose-dependently inhibited NKA-induced RAR stimulation did not significantly influence augmentation of the RAR and RL responses to SP. Administration of atropine (2 mg/kg, n = 6) in thiorphan-treated rabbits, which had no effect on NKA- and NKB-induced RAR stimuli, significantly attenuated the increases of RAR activity and RL induced by SP. These results suggest that tachykinin-induced RAR stimulation is mediated by the activation of NK2 receptors, probably involving participation of NK1 receptors. Furthermore, potentiation of the increases of RAR activity and RL produced by SP administration in the presence of thiorphan is partly mediated by facilitation of cholinergic neurotransmission. PMID- 9399973 TI - Heparinase III exerts endothelial and cardioprotective effects in feline myocardial ischemia-reperfusion injury. AB - The initial phase of neutrophil (PMN) adherence in the pathophysiology of myocardial ischemia-reperfusion (MI/R) injury depends on the selectins, particularly P- and L-selectin. Several ligands for these selectins have been identified, one of which may be a heparan sulfate proteoglycan (HSPG). Cats subjected to 90 min of MI and 270 min of R were given either heparinase III (0.033, 0.33 or 3.33 IU/kg/min) or its vehicle beginning 10 min before R and continuing throughout the 270-min R period. Heparinase III at 3.33 IU/kg/min provided a marked cardioprotective effect compared with cats receiving only vehicle as evidenced by a significant attenuation in myocardial necrosis (P < .01). In addition, endothelium-dependent vasorelaxation to acetylcholine in coronary artery rings isolated from MI/R cats treated with heparinase III was significantly preserved (P < .01). Adherence of PMNs to the coronary vascular endothelium after 270 min of R was also significantly attenuated in heparinase III-treated cats compared with vehicle (P < .01). At 0.33 IU/kg/min, heparinase III exerted modest, significant cardioprotective effects, whereas at 0.033 IU/kg/min, no significant beneficial effects were observed. Our results indicate that heparinase III is cardioprotective in a dose-dependent manner, preserves endothelial function and attenuates PMN adherence to the coronary vascular endothelium. PMID- 9399974 TI - Interactions of nonsteroidal anti-inflammatory drugs with rat renal organic anion transporter, OAT-K1. AB - We recently cloned and characterized the rat kidney-specific organic anion transporter, OAT-K1, which was suggested to mediate renal tubular transport of methotrexate. In this study, we investigated the interactions of nonsteroidal anti-inflammatory drugs (NSAIDs) with OAT-K1 by evaluating the effects of these drugs on renal distribution of methotrexate in vivo, and on methotrexate accumulation in the stably transfected LLC-PK1 cells expressing OAT-K1 (LLC-OAT K1). NSAIDs such as indomethacin and ketoprofen had significant inhibitory effects on renal accumulation of methotrexate in rats after coadministration. Indomethacin and ketoprofen inhibited methotrexate accumulation by LLC-OAT-K1 cells in a competitive manner with the apparent inhibition constant values of 1. 0 mM and 1.9 mM, respectively. Other NSAIDs including ibuprofen, flufenamate and phenylbutazone also showed potent inhibitory effects on methotrexate accumulation. However, indomethacin was not transported via OAT-K1. These results indicate that NSAIDs have potent inhibitory effects against the OAT-K1-mediated methotrexate transport, which suggests that the OAT-K1 may be one of interaction sites for methotrexate and NSAIDs in the kidney. PMID- 9399975 TI - Positive modulation of pepsinogen secretion by gastric acidity after vagal cholinergic stimulation. AB - Parallel increments of gastric acid and pepsinogen secretion generally occur after the application of cholinergic stimuli. However, it still remains to be established whether the changes in acid output associated with cholinergic stimulation play a role in regulation of the concomitant peptic secretory activity. In the present study, an anesthetized rat model was used for the evaluation of pepsinogen secretion in order to pursue a dual purpose: 1) to assess the relative functional relevance of direct and acid-dependent control exerted by cholinergic pathways on pepsinogen output; 2) to characterize the mechanisms through which changes in acidity within the stomach lumen may affect the peptic secretory activity of gastric mucosa. Bethanechol, 2-deoxy-D-glucose or electrical vagal stimulation caused parallel and atropine-sensitive increments of peptic and acid secretions. Omeprazole, a selective inhibitor of gastric H+:K+ adenosintriphosphatase, blocked the increase in acid but not pepsinogen secretion induced by bethanechol. However, 2-deoxy-D-glucose or electrical vagal stimulation failed to increase either pepsinogen or acid secretion in omeprazole pretreated rats. When tested in animals pretreated with both omeprazole and physostigmine (a drug able to prevent the enzymatic breakdown of vagally released ACh through the blockade of acetylcholinesterase), 2-deoxy-D-glucose or electrical vagal stimulation significantly increased pepsinogen secretion without affecting acid secretion. In omeprazole-pretreated rats, perfusion of the gastric lumen with acid solutions caused a pH-dependent and atropine-sensitive increase in peptic output only when applied in combination with electrical vagal stimulation. Functional ablation of capsaicin-sensitive sensory neurons did not modify the gastric secretory responses induced by bethanechol or electrical vagal stimulation. However, after topical application of lidocaine to the gastric mucosal surface, bethanechol stimulated both peptic and acid outputs, whereas electrical vagal stimulation only evoked acid secretion without affecting basal peptic output. The present results indicate that the activation of muscarinic receptors by vagally released ACh is not sufficient by itself to stimulate pepsinogen secretion and that a facilitatory action mediated by acid secretion is necessary to allow an increment of peptic output in response to vagal cholinergic stimuli. It is suggested that such facilitatory input is driven to chief cells by local intramural reflexes that involve capsaicin-insensitive intrinsic nerves. PMID- 9399976 TI - Pharmacokinetic-pharmacodynamic characterization of the cardiovascular, hypnotic, EEG and ventilatory responses to dexmedetomidine in the rat. AB - This study characterizes the pharmacokinetic-pharmacodynamic (PK-PD) relationships of the cardiovascular, EEG, hypnotic and ventilatory effects of the alpha-2 adrenergic agonist dexmedetomidine in rats. Dexmedetomidine was administered by a single rapid infusion (n = 6) and by an infusion regimen of gradually increasing rate (n = 8). HR, mean arterial pressure (MAP) and EEG signals were recorded continuously, as was the time at which the rats woke up spontaneously from drug-induced sleep, a measure of hypnosis. Arterial concentrations of dexmedetomidine and blood gases were determined regularly. A sigmoidal Emax model was used to describe the HR, MAP and EEG concentration effect relationships, with the EEG effect (activity in 0.5-3.5-Hz frequency band) linked to an effect-site model. The PK of dexmedetomidine could be described by a two-compartment model, with similar PK parameters for both infusion regimens. Plasma protein binding was 84.1[0.7]%. Because of complex cardiovascular homeostatic reflex mechanisms, HR and MAP could only be analyzed during gradually increasing infusions. The maximal decrease in HR was 35(2)%, and the maximal increase in MAP was 37(2)%. For both infusion regimens, similar PD parameters were found for the EEG and the hypnotic measure. These data suggest the absence of active metabolites or tolerance of the EEG and hypnotic effects. Judging on the basis of concentrations of dexmedetomidine (mean (S.E. M.)), HR decrease was the most sensitive response [EC50 of 0.65(0. 09) ng/ml], followed by increase in MAP [EC50 of 2.01(0.14) ng/ml], change in EEG activity [EC50 of 2.24(0.16) ng/ml] and the hypnotic measure [Cwake-up of 2.64(0.10) ng/ml]. Ventilatory effects were minor. PMID- 9399977 TI - SB 202026: a novel muscarinic partial agonist with functional selectivity for M1 receptors. AB - The finding that ascending cholinergic systems are severely degenerated in Alzheimer's disease has driven the search for a cholinomimetic therapy. Adverse effects observed with cholinesterase inhibitors and high-efficacy muscarinic agonists led us to design compounds with an improved profile. SB 202026 (R-(Z) (+)-alpha-(methoxyimino)-1-azabicyclo[2.2.2] octane-3-acetonitrile) displaced [3H]-oxotremorine-M from muscarinic receptors in the rat brain with high affinity (IC50 = 14 nM), a potency similar to that of oxotremorine-M itself (IC50 = 13 nM), but exhibited low affinity for cholinergic nicotinic receptors and other neuroreceptors. In studies using cloned human muscarinic receptors, SB 202026 possessed approximately equal affinity in displacing [3H]-quinuclidinyl benzilate from all muscarinic receptor subtypes. In functional models in vitro, SB 202026 caused maximal depolarization of the rat superior cervical ganglion at low concentrations (300 nM) (M1-mediated effect), while producing a lower maximal effect than the high-efficacy agonists oxotremorine-M and carbachol on M2 mediated release of ACh and M3-mediated smooth muscle contraction (guinea pig ileum), respectively. The functional selectivity and partial agonist profile seen in vitro were reflected in vivo through potent cognition-related activity (M1 induced increase in hippocampal EEG power) combined with low efficacy, compared with arecoline or oxotremorine, on induction of bradycardia (M2-mediated response), hypotension (via M3-mediated vasorelaxation) and tremor (thought to be mediated by M3 receptors). The foregoing profile of SB 202026 predicted that cognition-enhancing activity would be achieved at doses below those that initiate undesirable side effects, and this has subsequently been demonstrated in rodents, marmosets and humans. PMID- 9399978 TI - Inhibition of cyclooxygenase-2 rapidly reverses inflammatory hyperalgesia and prostaglandin E2 production. AB - PGs derived from cyclooxygenase-2 (COX-2), in particular PGE2, play important roles in the initiation of inflammation and pain. In the present study, we evaluated the role of COX-2-derived PGE2 in an animal model of established hyperalgesia. Inflammation and hyperalgesia were first induced by injection of carrageenan into rat footpads. Then we investigated the effects of subsequent therapeutic treatment with a selective COX inhibitor, with a nonsteroidal anti inflammatory drug and with anti-PGE2 antibody. Test compounds were administered 1 to 3 hr after carrageenan challenge, and inhibition of pain (hyperalgesia, measured by withdrawal from a thermal stimulus), and changes in paw edema and PG levels were evaluated. The i.v. administration of a nonselective COX inhibitor, ketorolac, caused a rapid reduction in hyperalgesia in the inflamed footpad, returning it to near-normal values within 1 hr. Normal (control) paw response times were not affected. Therapeutic administration of ketorolac prevented most further swelling caused by carrageenan but did not reverse edema already present at the time of dosing. Administered p.o., a selective COX-2 inhibitor (SC-58635) was as efficacious as ketorolac in reducing inflammatory hyperalgesia. Footpad PG levels returned to base line or below within 5 min of dosing with ketorolac, which suggests rapid turnover of PG in the inflamed tissue. Therapeutic treatment with a monoclonal anti-PGE2 antibody also fully reversed the hyperalgesia response. These studies suggest that continuous production of PGE2 by the COX-2 enzyme is a critical element in sustaining the hyperalgesic response at sites of tissue inflammation. PMID- 9399979 TI - Active immunization alters the plasma nicotine concentration in rats. AB - The ability of active immunization to alter nicotine distribution was studied in rats. Animals were immunized with 6-(carboxymethylureido)-(+/-)-nicotine (CMUNic) linked to keyhole limpet hemocyanin (KLH). Antibody titers determined by ELISA, using CMUNic coupled to albumin as the coating antigen, were greater than 1:10,000. Antibody binding was inhibited by neither of the nicotine metabolites cotinine and nicotine-N-oxide but was inhibited to a greater extent by CMUNic than by nicotine; this suggests the presence of antibodies to the linker structure as well as antibodies to nicotine. Antibody affinity for nicotine measured by soluble radioimmunoassay was 2.4 +/- 1.6 x 10(7) M-1, and binding capacity was 1.3 +/- 0.7 x 10(-6) M, which corresponds to 0.1 +/- 0.05 mg/ml of nicotine-specific IgG per milliliter of serum. One week after their second boost, groups of eight anesthetized rats immunized with either CMUNic-KLH or KLH alone received nicotine 0.03 mg/kg (equivalent to two cigarettes in a human) via the jugular vein over 10 sec. This dosing regimen was shown to mimic the arterio venous nicotine concentration gradient typical of nicotine delivered by cigarette smoking in humans. Plasma nicotine concentrations at 10 to 40 min were 4 to 6 fold higher in the CMUNic-KLH rats than in controls (P < .001). Nicotine binding in plasma determined by equilibrium dialysis was markedly increased in the CMUNic KLH group (83.4 +/- 6.8% vs. 16.4 +/- 14.2%), but brain nicotine concentrations at 40 min did not differ (37.9 +/- 4.5 vs. 44.0 +/- 8. 4 ng/g, CMUNic-KLH vs. KLH, P = .1). The amount of nicotine bound to antibody in plasma, estimated from the in vivo data, was 9% of the administered dose. These data demonstrate that active immunization can bind a significant fraction of a clinically relevant nicotine dose in plasma. Observing this effect with antibodies of modest affinity and titer is encouraging, but better immunogens may be needed to alter nicotine distribution to brain and modify nicotine's behavioral effects. PMID- 9399981 TI - Alcohol dehydrogenase-2*3 allele protects against alcohol-related birth defects among African Americans. AB - Considerable variation in offspring outcome is observed after intrauterine alcohol exposure. The underlying mechanism may include genetic diversity in the enzymes responsible for alcohol metabolism. Of the known genetic polymorphisms, differences at the alcohol dehydrogenase-2 locus (ADH2) are likely most critical because the resulting enzymes are >30-fold different in their kinetic constants. To test whether differences in maternal or offspring ADH2 genotype are determinants of risk for alcohol-related birth defects, maternal-infant pairs (n = 243) were enrolled on the basis of maternal alcohol intake during pregnancy and maternal ADH2 genotype. Infant outcome was measured using the Bayley Scales of Infant Development Mental Index (MDI) at 12 months of age. Drinking during pregnancy was associated with lower MDI scores but only in the offspring of mothers without an ADH2*3 allele (P < .01, analysis of variance, post hoc). The offspring of drinking women with at least one ADH2*3 allele had MDI scores similar to those of nondrinking women of either ADH2 genotype. Lower MDI scores were associated with the three-way interaction among increasing alcohol intake and maternal and offspring absence of the ADH2*3 allele (P < .01, multiple linear regression). We suggest that the protection afforded by this allele is secondary to its encoding of the high-Km/high-Vmax ADH beta3 isoenzyme, which would provide more efficient alcohol metabolism at high blood alcohol concentrations. These observations are supportive of alcohol, rather than acetaldehyde, being the more important proximate teratogen and are the first observations of a specific genetic explanation for susceptibility differences to alcohol-related birth defects. PMID- 9399980 TI - Angiotensin-converting enzyme inhibition and angiotensin II subtype-1 receptor blockade during the progression of left ventricular dysfunction: differential effects on myocyte contractile processes. AB - Inhibition of the angiotensin-converting enzyme (ACE) in the setting of chronic left ventricular (LV) dysfunction has been demonstrated to have beneficial effects on survival and symptoms. However, whether ACE inhibition has direct effects on myocyte contractile processes and if these effects are mediated primarily through the AT1 angiotensin-II receptor subtype remains unclear. The present project examined the relationship between changes in LV and myocyte function and beta adrenergic receptor transduction in four groups of six dogs each: (1) Rapid Pace: LV failure induced by chronic rapid pacing (4 weeks; 216 +/ 2 bpm); (2) Rapid Pace/ACEI: concomitant ACE inhibition (ACEI: fosinopril 30 mg/kg b.i.d.) with chronic pacing; (3) Rapid Pace/AT1 Block: concomitant AT1 Ang II receptor blockade [Irbesartan: SR 47436(BMS-186295) 30 mg/kg b.i.d.] with chronic pacing; and (4) CONTROL: sham controls. With Rapid Pace, the LV end diastolic volume increased by 62% and the ejection fraction decreased by 53% from control. With Rapid Pace/ACEI, the LV end-diastolic volume was reduced by 24% and the ejection fraction increased by 26% from Rapid Pace only values. Rapid Pace/AT1 Block did not improve LV geometry or function from Rapid Pace values. Myocyte contractile function decreased by 40% with Rapid Pace and increased from this value by 32% with Rapid Pace/ACEI. Rapid Pace/AT1 Block had no effect on myocyte function when compared with Rapid Pace values. With Rapid Pace/ACEI, beta receptor density and cyclic AMP production were normalized and associated with an improvement in myocyte beta adrenergic response compared with Rapid Pace only. Although Rapid Pace/AT1 also normalized beta receptor density, cyclic AMP production was unchanged and myocyte beta adrenergic response was reduced by 15% compared with Rapid Pace only. ACE inhibition with chronic rapid pacing improved LV and myocyte geometry and function, and normalized beta receptor density and cyclic AMP production. However, AT1 Ang-II receptor blockade with chronic rapid pacing failed to provide similar protective effects on LV and myocyte geometry and function. These unique findings suggest that the effects of ACE inhibition on LV geometry and myocyte contractile processes in the setting of developing LV failure are not primarily caused by modulation of AT1 Ang-II receptor activation. PMID- 9399982 TI - Absence of tachykinin involvement in leukotriene D4 and antigen-induced contraction of guinea pig isolated bronchus. AB - In guinea pig airways, contractions induced by leukotriene D4 or antigen are thought to be mediated primarily by an action of the agonist or of released mast cell-derived mediators directly on the airway smooth muscle cell. An indirect contractile action mediated by endogenous tachykinins also has been described for both of these stimuli. The present study evaluated the contribution of endogenous tachykinins to ovalbumin- and leukotriene D4-induced contractions in the guinea pig bronchus by modulating the concentrations of tachykinins within the tissues and by using neurokinin receptor antagonists. Acute depletion of tachykinins with capsaicin had no effect on responses elicited by either stimulus. Similarly, tetrodotoxin treatment failed to influence leukotriene D4-induced contractions. Inhibitors of neutral endopeptidase (thiorphan) and angiotensin-converting enzyme (lisinopril) enhanced neurally mediated tachykininergic responses and potentiated leukotriene D4. The latter effect persisted in the presence of tetrodotoxin or the neurokinin antagonists CP99994 and SR48968 and in tissues treated acutely with capsaicin. The potentiation was absent, however, from bronchi incubated with L-cysteine. Ovalbumin-induced contractions were unaltered by inhibition of neutral endopeptidase and angiotensin-converting enzyme. These observations suggest that tachykinins are not involved in mediation of leukotriene D4- or antigen-induced contractions of the guinea pig bronchus. The ability of protease inhibitors to potentiate leukotriene D4 but not antigen-induced responses is therefore ascribed to inhibition of bioinactivation of leukotriene D4 to leukotriene E4. PMID- 9399983 TI - Ro 61-1790, a new hydrosoluble endothelin antagonist: general pharmacology and effects on experimental cerebral vasospasm. AB - Endothelin (ET) receptor antagonists are of great potential clinical interest for the treatment pathological conditions associated with vasospasm, such as subarachnoid hemorrhage (SAH). We developed for parenteral use a compound of a class of trifunctionalized heteroarylsulfonamide pyrimidines specially designed for high water solubility. Ro 61-1790 [5-methyl-pyridine-2-sulfonic acid 6-(2 hydroxy-ethoxy)-5-(2-methoxy-phenoxy)-2-(2-1H-tetrazol-5-yl-+ ++pyri din-4-yl) pyrimidin-4-ylamide] is a competitive ET antagonist with an affinity to ETA receptor in the subnanomolar range. It has a approximately 1000-fold selectivity for the ETA vs. the ETB receptor as assessed on functional assays (e.g., ET-1 induced inositol-1,4, 5-triphosphate release or ET-1-induced intracellular calcium mobilization). Ro 61-1790 also had a high functional potency for inhibiting contraction induced by ET-1 on isolated rat aorta (ETA receptors; pA2 = 9.5) or by sarafotoxin S6c on rat trachea (ETB receptors; pA2 = 6.4). In vivo, Ro 61-1790 inhibited the pressor effect of big ET-1 in pithed rats with an ID50 value of 0.05 mg/kg. Intravenous bolus of Ro 61-1790 induced a long-lasting antihypertensive effect in deoxycorticosterone acetate salt rats instrumented with telemetry. In a double-hemorrhage canine model of SAH, Ro 61-1790 both prevented and reversed cerebral vasospasm in a dose-dependent manner. In an established cerebral vasospasm, 3 mg/kg Ro 61-1790 i.v. was half as efficacious as intrabasilar papaverine. Ro 61-1790 (20 mg/kg/day) totally prevented the occurrence of vasospasm. In summary, these data demonstrate that Ro 61-1790 is a potent and selective ETA receptor antagonist suitable for parenteral use and potentially useful for preventing delayed ischemic deficit in patients with SAH. PMID- 9399984 TI - Pharmacokinetic-pharmacodynamic modeling of stimulatory and sedative effects of alprazolam: timing performance deficits. AB - Alprazolam decreased the reinforcement rate and increased the shorter-response rate of contingency-controlled timing behavior under a differential reinforcement of low-rate schedule (DRL 45-s) in rats. An integrated pharmacokinetic pharmacodynamic (PK-PD) model was developed to describe and characterize the effects of i.v. and s. c. administration of alprazolam. The onset, peak and disappearance of alprazolam effects were evaluated during a 3-hr session. After s. c. alprazolam administration, two peak increases in shorter-response rate occurred at moderate alprazolam serum levels, first in the ascending and then in the descending limb of the concentration-time profile. We used a stimulation sedation PD model incorporating two opposing effect-link sigmoidal Emax functions to model the two peaks after s.c. alprazolam administration. The model suggested that alprazolam possesses both stimulatory and sedative effects in a continuous but sequential fashion, which corresponded to low- and high-concentration effects as indicated by the EC50 values of 0.09 and 0.18 microg/ml, respectively. Owing to the rapid onset of i.v. administration, the first peak (a transition phase before the onset of the sedative effect) was absent, with the presence of the second peak again coinciding with the offset of the sedative effect. The reinforcement rate (IC50 = 0.02 microg/ml) characterized by the indirect response model to account for the initial hysteresis is an index for evaluating the deficit in timing performance. Although the effects of alprazolam can be described in behavioral terms, simultaneous PK-PD optimization numerically defines the performance and hypothesizes the coexistence of stimulation and sedation components for alprazolam. The stimulation-sedation model may help in delineating the possible mechanisms for adverse rebound side effects and of tolerance in humans. PMID- 9399985 TI - Nonpeptide endothelin receptor antagonists. X. Inhibition of endothelin-1- and hypoxia-induced pulmonary pressor responses in the guinea pig by the endothelin receptor antagonist, SB 217242. AB - This study investigated the effects of the nonpeptide endothelin (ET) receptor antagonist, SB 217242, against ET-1-induced pulmonary pressor responses and in a model of hypoxia-induced pulmonary hypertension in the guinea pig. In guinea pig isolated pulmonary artery rings, SB 217242 (3-300 nM) produced a concentration dependent inhibition of ET-1-induced contractions, with a pA2 of 8.1. SB 217242 (1 or 3 mg/kg i.v.) elicited a dose-related inhibition of ET-1-induced increases in pulmonary artery and airway insufflation pressure responses in anesthetized guinea pigs. Chronic exposure to hypoxia (9% O2 for 0-14 days) produced a time dependent increase in mean pulmonary artery pressure. After a 10-day exposure to hypoxia there was about a 100% elevation in pulmonary artery pressure, and right ventricular mass and plasma irET levels increased 3-fold compared with normoxic animals. SB 217242, administered by continuous intraperitoneal infusion via mini osmotic pump (0.36, 3.6 or 10.8 mg/day), significantly reduced (by about 50%) hypoxia-induced pulmonary artery pressure increases at all three doses used. The hypoxia-induced right ventricular hypertrophy was significantly attenuated by the 3.6 and 10.8 mg/day doses. Based on hematocrit, hemoglobin and red blood cell counts, SB 217242 did not affect the normal physiological erythropoietic response to hypoxia. There were no appreciable differences in the maximum contractile effects of ET-1 or the potency of SB 217242 (pKB values, 8.3 and 8.0, respectively) versus ET-1-induced responses in isolated pulmonary arteries from hypoxic versus normoxic guinea pigs. However, there was a marked reduction in endothelium-dependent relaxation of precontracted pulmonary artery isolated from hypoxic compared with normoxic animals. The results of the present study provide further preclinical evidence for a pathophysiological role of ET-1 and the potential therapeutic utility of ET receptor antagonists, such as SB 217242, in pulmonary hypertension. PMID- 9399987 TI - Elevated environmental temperatures can induce hyperthermia during d-fenfluramine exposure and enhance 5-hydroxytryptamine (5-HT) depletion in the brain. AB - d-Fenfluramine (d-Fen) has been demonstrated to alter body temperature (BT), decrease 5-hydroxytryptamine (5-HT) and decrease 5-HT plasma membrane transporters (PMT) in rats. Therefore, experiments were designed to test whether a correlation existed between elevated BT and brain 5-HT depletions. It was hypothesized that d-Fen would induce hyperthermia if the environmental temperature was elevated. Experiments were conducted to determine 1) the dose response of d-Fen on BT in a 28 degrees C environment, 2) the acute effect of d Fen on long-term depletion of 5-HT and 5-HT PMT in a 4 degrees C, 22 degrees C or 28 degrees C environment and 3) the effect of a 22 degrees C environment vs. a 28 degrees C environment on the plasma levels of d-Fen and d-norfenfluramine. d-Fen produced a dose-dependent elevation of BT in the 28 degrees C environment, decreased BT in the 4 degrees C environment and had no effect on BT in the 22 degrees C environment. Exposure to d-Fen in the 4 degrees C or 22 degrees C environment reduced 5-HT and 5-HT PMT concentrations compared with control. However, greater reductions of 5-HT and 5-HT PMT concentrations occurred in the 28 degrees C environment. Conversely, the plasma levels of d-Fen and d norfenfluramine were not altered. Thus these experiments demonstrate that increased BT during d-Fen exposure occurs at elevated environmental temperatures without altering the plasma concentrations of the drug and results in an enhanced long-term depletion of brain 5-HT and 5-HT PMT. PMID- 9399986 TI - The effect of the enzyme inhibitor phenylmethylsulfonyl fluoride on the pharmacological effect of anandamide in the mouse model of cannabimimetic activity. AB - Anandamide is an putative endogenous cannabinoid ligand that produces pharmacological effects similar to those of Delta9-tetrahydrocannabinol, the principle psychoactive constituent in marijuana. There is considerable evidence that the enzyme inhibitor phenylmethylsulfonyl fluoride (PMSF) is capable of altering the actions of anandamide in vitro by blocking its metabolism. Therefore, studies were conducted in mice to determine whether PMSF could produce cannabinoid effects by altering endogenous levels of anandamide as well as determining whether PMSF could potentiate the effects of exogenously administered anandamide. Mice receiving i.p. injections of PMSF exhibited cannabinoid effects that included antinociception, hypothermia and immobility with ED50 values of 86, 224 and 206 mg/kg, respectively. Spontaneous activity was reduced at doses greater than 100 mg/kg. However, none of these effects was blocked by the cannabinoid antagonist SR 141716A. On the other hand, pretreatment with an inactive dose of PMSF (30 mg/kg) potentiated the effects of anandamide on tail flick response (antinociception), spontaneous activity and mobility by 5-, 10- and 8-fold, respectively. PMSF did not alter anandamide's hypothermic effects. Overall, these findings with PMSF underscore the importance of metabolism in the actions of anandamide. It still must be established whether metabolites of anandamide contribute to its pharmacological activity. PMID- 9399988 TI - Blood-brain disposition and antinociceptive effects of -D-penicillamine2,5 enkephalin in the mouse. AB - Although intravenous administration of [D-penicillamine2, 5]-enkephalin (DPDPE) produces significant antinociception in rodents, the duration of antinociception is short ( approximately 15 min). The present study was conducted to test the hypothesis that duration of antinociception for DPDPE is determined by both systemic and regional disposition (i.e., blood-brain translocation), and that the magnitude of antinociception is related more closely to concentrations in brain tissue than in blood. Systemic disposition was examined after i.v. administration of DPDPE (10-100 mg/kg) to male CD-1 mice. The relationship between antinociception and concentration in blood and brain tissue was assessed by determining antinociception 10 min after administration of DPDPE (10-100 mg/kg); effect versus brain tissue concentration data were fit with pharmacodynamic models to recover EC50 estimates. In addition, the time course of antinociception, as well as blood and brain tissue concentrations, were examined after an i.v. bolus dose (40 mg/kg) of DPDPE. The systemic disposition of DPDPE was nonlinear; both clearance and volume of distribution were dose-dependent. Antinociception increased proportionately with increasing concentrations of DPDPE in blood or brain tissue, with an EC50 of 1.42 +/- 0.06 microg/g expressed as brain tissue concentration. However, the brain-to-blood concentration ratio also increased with increasing dose, suggestive of saturable translocation of DPDPE across the blood-brain barrier. Antinociception appeared rapidly (within 5 min) and dissipated within approximately 15 min after a 40 mg/kg i.v. dose. These results suggest that rapid elimination from blood and active efflux from brain limit the duration of action of DPDPE. PMID- 9399989 TI - Synergistic effects of cocaine with lateral hypothalamic brain stimulation reward: lack of tolerance or sensitization. AB - The curve-shift (rate-frequency) paradigm was used to quantify the interaction of cocaine administration with the rewarding effects of lateral hypothalamic electrical stimulation. First, eight animals were tested at 48-h intervals with increasing doses of cocaine (0.5, 1, 2, 4, 8, 16 or 32 mg/kg i.p.); tests with saline were given on intervening days. Cocaine produced dose-orderly leftward shifts of the functions relating response rate to stimulation frequency, which reduced, for each animal, the amount of stimulation required to sustain responding; the two highest doses of the drug shifted the mean rate-frequency curve by 0.47 log units, more than doubling the rewarding potency of the brain stimulation. Baseline thresholds did not change between tests. Next, evidence for sensitization or tolerance was sought from five additional groups of animals, one group given 4 mg/kg and two groups given 16 mg/kg of cocaine at 48-h intervals, and another two groups maintained for 7 days with thrice-daily injections of 10 mg/kg of cocaine or saline. Consistent with results seen in other brain stimulation reward paradigms, there was no evidence of tolerance or sensitization to cocaine's reward-potentiating effects as quantified in the rate-frequency paradigm. PMID- 9399990 TI - The In vitro hepatic metabolism of quinine in mice, rats and dogs: comparison with human liver microsomes. AB - The major metabolic pathway of quinine in the human has been shown to be 3 hydroxylation mediated mainly by human cytochrome P450 (CYP) 3A4. In this extended in vitro study, quinine 3-hydroxylation was further investigated using microsomes from mouse, rat, dog and human livers and was compared among them in terms of the in vitro enzyme-kinetic parameters and quinine-drug interaction screenings. In all species, 3-hydroxyquinine was the principal metabolite of quinine. There was intra- and interspecies variability among all the kinetic parameters, and dogs exhibited a closer resemblance to humans in terms of the mean kinetic data. Ketoconazole and troleandomycin inhibited the 3-hydroxylation of quinine in all species. Both alpha-naphthoflavone and diazepam showed an interspecies difference in quinine 3-hydroxylation: a trend toward an activation in dog and human, and a significant inhibition in mouse and rat, liver microsomes. Antisera raised against rat CYP3A2 strongly inhibited quinine 3 hydroxylation by about 96, 84 and 92% with mouse, rat and dog liver microsomes, respectively, but neither anti-rat 2C11 and 2E1 antisera did so with rat liver microsomes. Primaquine, doxycycline and tetracycline substantially inhibited the formation of 3-hydroxyquinine in rat, dog and human species, but proguanil had no such effect in any species. Chloroquine inhibited quinine 3-hydroxylation with rat and dog liver microsomes but not with human liver microsomes. There was a significant correlation (r = 0.986, P < .001) between the CYP3A contents and the formation rates of 3-hydroxyquinine in eight human liver microsomal samples. It is concluded that 3-hydroxyquinine is a main metabolite of quinine and that CYP3A/Cyp3a is a principal isoform involved in this metabolic pathway in the respective (rat, dog and human/mouse) species tested. The dog and possibly the rat may be qualitatively and quantitatively suitable animal models for exploring the quinine 3-hydroxylase activity and for screening quinine-drug interactions in vitro, at certain inconsistency with the human liver microsomal data. PMID- 9399991 TI - Calpains mediate calcium and chloride influx during the late phase of cell injury. AB - The role of Ca++ in cell death is controversial. Extracellular Ca++ influx and calpain activation occurred during the late phase of renal proximal tubule cell injury produced by the mitochondrial inhibitor antimycin A. Chelation of intracellular Ca++, extracellular Ca++, the calcium channel blocker nifedipine, calpain inhibitor 1 and the dissimilar calpain inhibitor PD150606 blocked antimycin A-induced influx of extracellular Ca++ and cell death. The calcium channel blocker verapamil was ineffective. Calpain inhibitor 1 and PD150606 were cytoprotective also against tetrafluoroethyl-L-cysteine-, bromohydroquinone-, oxidant (t-butylhydroperoxide)- and calcium ionophore (ionomycin)-induced cell death. Extracellular Ca++ influx was associated with the translocation of calpain activity from the cytosol to the membrane and was prevented by calpain inhibitor 1, PD150606 and nifedipine. Finally, nifedipine, calpain inhibitor 1, PD150606 and the Cl- channel inhibitors [5-nitro-2-(3-phenylpropylamino)-benzoate, niflumic acid, diphenylamine-2-carboxylate, and indanyloxyacetic acid] blocked the increase in Cl- influx that occurs during the late phase of cell injury and triggers terminal cell swelling and death. These data suggest that Ca++ and calpains play a common and critical role in renal proximal tubule cell death produced by diverse agents. In addition, calpain activation appears to play a dual role during the late phase of cell injury. Initial calpain activation elicits extracellular Ca++ influx through a nifedipine-sensitive pathway, resulting in calpain translocation to the membrane and in turn Cl- influx. PMID- 9399992 TI - Modulation of nociception by microinjection of delta-1 and delta-2 opioid receptor ligands in the ventromedial medulla of the rat. AB - In this study, we characterized the role of delta-1 and delta-2 opioid receptors in the ventromedial medulla (VMM) in the modulation of thermal nociception. Male Sprague-Dawley rats were prepared with an intracerebral guide cannula aimed at the nucleus raphe magnus or nucleus reticularis gigantocellularis pars alpha. Microinjection of the delta-1 opioid receptor agonist [D-Pen2,D-Pen5]enkephalin (DPDPE) or the delta-2 opioid receptor agonist [D-Ala2, Glu4]deltorphin (DELT) in the VMM increased response latency in the radiant heat tail-flick test with respective ED50 values (95% CL) of 0.66 (0.07-1.5) nmol and 0.1 (0.03-0.21) nmol. In the 55 degrees C hot-plate test, DELT produced a modest, transient increase in response latency and DPDPE was ineffective. The antinociception produced by DPDPE was antagonized by microinjection at the same site of 1.5 pmol of the delta-1 opioid receptor antagonist 7-benzylidenenaltrexone (BNTX) but not by 0.15 nmol of the delta-2 opioid receptor antagonist naltriben (NTB). Conversely, the antinociception produced by DELT was antagonized by microinjection at the same site of 0.15 nmol of NTB but not by 1.5 pmol of BNTX. These doses of BNTX or NTB alone did not alter either tail-flick or hot-plate latency when microinjected in the VMM. However, at 10-fold higher doses, BNTX lost its selectivity for the delta-1 opioid receptor, and NTB by itself increased tail-flick and hot-plate latencies. These results collectively implicate both delta-1 and delta-2 opioid receptors in the VMM in the modulation of nociception. They also indicate that the antinociceptive effects of DPDPE and DELT can be distinguished by BNTX and NTB, providing additional support for the existence of delta-1 and delta-2 opioid receptor subtypes at supraspinal loci. Finally, the failure of effective doses of either BNTX or NTB to alter nociceptive threshold suggests that neurons in the VMM do not receive a tonic, inhibitory enkephalinergic input mediated by delta-1 or delta-2 receptors. PMID- 9399993 TI - Inhibition of guinea pig detrusor contraction by NS-1619 is associated with activation of BKCa and inhibition of calcium currents. AB - The effects of NS-1619 on bladder contractile function and on transmembrane currents were evaluated in vitro on isolated guinea pig detrusor strips and isolated detrusor myocytes, respectively. In the isolated bladder strip, NS-1619 inhibited KCl-induced contractions in a concentration-dependent manner (IC50 = 12.2 +/- 3. 2 microM). Isolated detrusor myocytes were quiescent and had resting membrane potentials that averaged -45.3 +/- 2.7 mV. With patch-clamp techniques we demonstrated that exposure to 10 to 100 microM NS-1619 increased an iberiotoxin-sensitive current consistent with the activation of the large conductance calcium-dependent potassium channel (BKCa). Single-channel analysis confirmed that NS-1619 increased the open probability of BKCa channels. NS-1619 also appeared to decrease inward calcium current (ICa). After exposure to 30 microM NS-1619, peak current amplitude significantly decreased by approximately 50%. Analysis of the current voltage relationship revealed a significant decrease in maximal conductance from 10.5 +/- 4 to 6.2 +/- 3 nS. The voltage dependence of calcium current activation and inactivation was well fit by a Boltzmann relationship. Besides the decrease in conductance, there was a small, but significant shift in the half-inactivation voltage, which suggests that NS-1619 preferentially blocks the open state of the channel. Steady-state (window) calcium current was also decreased. Analysis of the theoretical window current revealed a 71% decrease in this noninactivating current. These data indicate that NS-1619 inhibits detrusor smooth muscle contraction in a concentration-dependent manner and that the underlying mechanism of action for this effect involves inhibition of calcium current, and may also include activation of the BKCa channel. Compounds with this profile may be useful in the treatment of bladder instability. PMID- 9399994 TI - Inhibition of neutrophil elastase in CF sputum by L-658,758. AB - Elastases in cystic fibrosis (CF) pulmonary fluids damage lung tissue and perpetuate cycles of infection, inflammation and injury. Elastases from three different sources may be present in CF airways: neutrophils, macrophages and Pseudomonas. We measured how well the cephalosporin-based antielastase L-658,758 blocks the activity of human neutrophil elastase (NE), human proteinase-3, human macrophage metalloelastase, mouse macrophage metalloelastase and Pseudomonas aeruginosa elastase. We also examined the ability of L-658,758 to block elastases in CF sputum in vitro. Sputum samples from adult CF patients were fractionated to obtain the aqueous sol phase. These were then studied individually or pooled. Elastinolytic activity, which ranged from 3.2 microg elastin degraded/ml sol/min to 26.3 microg elastin degraded/ml sol/min, was measurable in every individual sol sample and in the pooled sol. L-658,758 effectively inhibited elastinolysis by NE, proteinase-3 and the pooled sol but did not inhibit the activity of the metalloelastases, human and mouse macrophage metalloelastase and Pseudomonas elastase. Secretory leukoprotease inhibitor, which inhibited NE but did not inhibit proteinase-3, blocked 90% of sol elastinolytic activity; this suggests that the majority of this activity in the pooled sol derived from NE. L-658,758 was an effective inhibitor of sol elastase, blocking more than 97% of elastinolytic activity in the individual sol samples. We conclude that L-658,758 is an effective inhibitor of NE, proteinase-3 and CF sputum sol elastase. PMID- 9399995 TI - Pharmacological characterization of novel cyanoguanidines as vascular KATP channel blockers. AB - KATP blockers derived from cyanoguanidine KATP opener (P1075) chemistry were characterized in isolated rabbit mesenteric artery and evaluated functionally by their ability to antagonize maximal relaxation induced by pinacidil (1 microM) of norepinephrine (5 microM) contraction. PNU-89692, PNU-97025E and PNU-99963 were identified as KATP blockers with IC50 values of 860, 83 and 18 nM, respectively. Studies with selected chiral compounds demonstrated that the (R)-enantiomers were more potent as KATP blockers than the (S)-enantiomers. Further studies demonstrated that PNU-99963 (1) inhibited relaxations by other KATP openers, such as cromakalim (0.5 microM) and minoxidil sulfate (5 microM); (2) was more potent than the other known vascular KATP blockers (glyburide and PNU-37883A); and (3) acted as a KATP blocker in isolated rat aorta as well as dog coronary artery. PNU 99963 actions were selective because PNU-99963 (100 nM) was without any inhibitory effect on relaxations induced by forskolin (0.5 microM), nitroglycerin (1 microM), D600 (25 or 500 nM) or 15 mM K+-induced relaxations of NE contractions in K+-free PSS. The discovery of KATP blockers and openers from the same chemical series is a first for the K+ channel field. The close structural similarity between P1075 (KATP opener) and PNU-99963 (KATP blocker), stereospecificity of action and potency and selectivity all suggest that these molecules may prove to be valuable tools in understanding the structure and function of the KATP channel complex in vascular smooth muscle. PMID- 9399996 TI - Chronic ethanol exposure leads to a selective enhancement of N-methyl-D-aspartate receptor function in cultured hippocampal neurons. AB - Effects of chronic ethanol exposure on N-methyl-D-aspartate (NMDA) receptor function were examined in hippocampal neurons. Rat hippocampal neurons grown in culture were chronically exposed to 100 mM ethanol to examine mechanisms that could underlie ethanol-induced changes in receptor function and excitotoxicity. NMDA-stimulated, but not kainic acid-stimulated, increases in intracellular calcium were enhanced after 1-, 2- and 7-day exposures to 100 mM ethanol. Chronic exposure to ethanol for 7 days duration increased the magnitude of cell death mediated by NMDA application, but not that mediated by alpha-amino-3-hydroxy-5 methylisoxazole-4-propionate or kainic acid exposure. In addition, NMDA-induced excitotoxicity after chronic ethanol exposure (CEE) was not altered in the presence of nifedipine. The enhancement of NMDA-induced neuronal cell death was evident after 2 days of CEE, but not significantly different after a 1-day exposure to 100 mM ethanol. The enhancement of NMDA-induced calcium responses and excitotoxicity could be mimicked by a chronic 7-day exposure to aminophosphonovaleric acid. However, a concomitant chronic exposure of ethanol/aminophosphonovaleric acid did not enhance NMDA-induced calcium responses or excitotoxicity. Chronic exposure paradigms did not consistently alter basal intracellular calcium levels nor total cell number in the absence of exposure to glutamate receptor agonist. These findings support the hypothesis that NMDA receptor function is enhanced after CEE, and this predisposes hippocampal neurons to excitotoxicity. PMID- 9399997 TI - Interaction of Alkyl/Arylphosphonates, phosphonocarboxylates and diphosphonates with different anion transport systems in the proximal renal tubule. AB - Luminal and contraluminal stop-flow microperfusion was applied, and the apparent Ki values (mmol/l) against the luminal phosphate and the contraluminal p aminohippurate (PAH), sulfate and dicarboxylate transport systems were evaluated. Luminal phosphate transporter: Among the 20 compounds tested only phosphonoformate (foscarnet), etidronate, and clodronate have a good affinity (app.Ki < 1 mmol/l), whereas the 2-naphthylphosphonates, phosphonoacetate, pamidronate, alendronate and aminomethanediphosphonates have a moderate affinity (app.Ki, 1.6-6.0 mmol/l). The other compounds tested had a low (app. Ki > 6 mmol/l) or no affinity. Contraluminal PAH transporter: The hydrophobic phenyl-, benzyl- or 2-naphthylphosphonates have good to moderate affinity, whereas the less hydrophobic alkylphosphonates, the phosphonocarboxylates (except 4 phosphonobutyrate) and all tested diphosphonates show no interaction. Sulfate transporter: 2-Naphthylmethylphosphonate and 2-naphthylmethyldifluorophosphonate have a good affinity (app.Ki 100 microM) were required to antagonize alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid-induced currents. They were potent, systemically active NMDA receptor antagonists in vivo against responses of single neurons in the rat spinal cord to microelectrophoretic application of NMDA with ID50 values in the low milligram per kilogram i.v. range. They also inhibited pentylenetetrazol-, NMDA- and maximal electroshock induced convulsions in mice with ED50 values ranging from 8 to 100 mg/kg i.p. The duration of anticonvulsive action was rather short but was prolonged by the organic acid transport inhibitor probenecid (200 mg/kg). The agents tested represent a novel class of systemically active glycineB antagonists with greatly improved bioavailability. PMID- 9400003 TI - Delta opioid modulation of the binding of guanosine-5'-O-(3 [35S]thio)triphosphate to NG108-15 cell membranes: characterization of agonist and inverse agonist effects. AB - The ability of the delta opioid agonist DPDPE ([D-Pen2, D-Pen4]enkephalin) to stimulate binding of the GTP analog guanosine-5'-O-(3-[35S]thio)triphosphate ([35S]GTPgammaS) to pertussis toxin-sensitive G proteins has been characterized in membranes from NG108-15 mouse neuroblastoma X rat glioma cells. The presence of GDP, or its hydrolysis-resistant analog GDPbetaS, and Mg++ ions was essential to observe agonist-mediated stimulation of [35S]GTPgammaS binding, although the guanine dinucleotides alone had complex inhibitory and stimulatory effects on [35S]GTPgammaS binding. The relative ability of the delta antagonists benzylidenenaltrexone and naltriben to inhibit DPDPE-stimulated [35S]GTPgammaS binding suggested the opioid receptor involved was of the delta-2 subtype. Ligand binding assays demonstrated biphasic binding of these antagonists to this single receptor type. [35S]GTPgammaS binding was also stimulated by [D Ser2,Leu5,Thr6]enkephalin > deltorphin II = DPDPE = etorphine > levallorphan = diprenorphine = nalorphine = naltrindole. The delta antagonists benzylidenenaltrexone, TIPP (Tyr-Tic-Phe-Phe) and naltriben had no effect, but ICI 174864 (N, N-diallyl-Tyr-Aib-Phe-Leu-OH) acted as an inverse agonist and inhibited [35S]GTPgammaS binding. Pertussis toxin pretreatment blocked agonist stimulation of [35S]GTPgammaS binding and also reduced basal binding, thus confirming the presence of constitutively active delta receptors. Replacement of Na+ in the assay buffer with K+ afforded an increased level of basal [35S]GTPgammaS binding and an apparent increase in both the inverse agonist activity of ICI 174864 and the agonist activity of the partial agonist diprenorphine relative to the full agonist [D-Ser2, Leu5,Thr6]enkephalin. The stimulation of [35S]GTPgammaS binding to NG108-15 cell membranes allows a functional measure of delta opioid activity that can provide systems of differing relative efficacy. PMID- 9400004 TI - Ro 25-6981, a highly potent and selective blocker of N-methyl-D-aspartate receptors containing the NR2B subunit. Characterization in vitro. AB - The interaction of Ro 25-6981 with N-methyl-D-aspartate (NMDA) receptors was characterized by a variety of different tests in vitro. Ro 25-6981 inhibited 3H MK-801 binding to rat forebrain membranes in a biphasic manner with IC50 values of 0.003 microM and 149 microM for high- (about 60%) and low-affinity sites, respectively. NMDA receptor subtypes expressed in Xenopus oocytes were blocked with IC50 values of 0.009 microM and 52 microM for the subunit combinations NR1C & NR2B and NR1C & NR2A, respectively, which indicated a >5000-fold selectivity. Like ifenprodil, Ro 25-6981 blocked NMDA receptor subtypes in an activity dependent manner. Ro 25-6981 protected cultured cortical neurons against glutamate toxicity (16 h exposure to 300 microM glutamate) and combined oxygen and glucose deprivation (60 min followed by 20 h recovery) with IC50 values of 0.4 microM and 0.04 microM, respectively. Ro 25-6981 was more potent than ifenprodil in all of these tests. It showed no protection against kainate toxicity (exposure to 500 microM for 20 h) and only weak activity in blocking Na+ and Ca++ channels, activated by exposure of cortical neurons to veratridine (10 microM) and potassium (50 mM), respectively. These findings demonstrate that Ro 25-6981 is a highly selective, activity-dependent blocker of NMDA receptors that contain the NR2B subunit. PMID- 9400005 TI - Acute experimental esophagitis activates a second signal transduction pathway in cat smooth muscle from the lower esophageal sphincter. AB - In single cells, isolated by enzymatic digestion from the circular muscle layer of the lower esophageal sphincter (LES), acute experimental esophagitis (AE) alters signal transduction in response to a maximally effective dose of acetylcholine. In normal LES contraction was inhibited by M3 >> M1 or M2 antagonists. In AE inhibition by M2 antagonists increased significantly so that contraction was inhibited by M3 > M2 > M1 antagonists. In normal cells permeabilized by saponin, contraction was antagonized by antibodies against Gq/11, by the phosphatidylinositol-specific phospholipase C (PI-PLC) antagonist U 73122, but not by the phosphatidylcholine-specific phospholipase C (PC-PLC) inhibitor D609, or by the phospholipase D pathway inhibitor propranolol. In AE contraction was reduced by Gq/11 and Gi3 antibodies and by U73122, propranolol and D609. After thapsigargin treatment of normal cells to reduce intracellular Ca++ stores, contraction was inhibited by M2 and M3 antagonists, by antibodies against Gq/11 and Gi3, by U73122, D609 and propranolol, suggesting that depletion of Ca++ stores reproduces the changes induced by AE. We conclude that in normal LES smooth muscle cells acetylcholine-induced contraction is mediated by M3 receptors linked to Gq/11 and PI-PLC, whereas in AE, contraction through this pathway is reduced, perhaps because of reduction in Ca++ stores, and a second pathway is activated by M2 receptors linked to Gi3, PC-PLC and phospholipase D. PMID- 9400006 TI - Neurotransmitter receptor and transporter binding profile of antidepressants and their metabolites. AB - Several new antidepressants that inhibit the serotonin (SERT) and norepinephrine transporters (NET) have been introduced into clinical practice the past several years. This report focuses on the further pharmacologic characterization of nefazodone and its metabolites within the serotonergic and noradrenergic systems, in comparison with other antidepressants. By use of radioligand binding assays, we measured the affinity (Ki) of 13 antidepressants and 6 metabolites for the rat and human SERT and NET. The Ki values for eight of the antidepressants and three metabolites were also determined for the rat 5-HT1A, 5-HT2A and muscarinic cholinergic receptors, the guinea pig histamine1 receptor and the human alpha-1 and alpha-2 receptors. These data are useful for predicting side effect profiles and the potential for pharmacodynamic drug-drug interactions of antidepressants. Of particular interest were the findings that paroxetine, generally thought of as a selective SERT antagonist, possesses moderately high affinity for the NET and that venlafaxine, which has been described as a "dual uptake inhibitor", possesses weak affinity for the NET. We observed significant correlations in SERT (r = 0.965) or NET (r = 0.983) affinity between rat and human transporters. Significant correlations were also observed between muscarinic cholinergic and NET affinity. There are several significant correlations between affinities for the 5-HT1A, 5-HT2A, histamine1, alpha-1 and alpha-2 receptors. These novel findings, not widely described previously, suggest that many of the individual drugs studied in these experiments possess some structural characteristic that determines affinity for several G protein-coupled, but not muscarinic, receptors. PMID- 9400007 TI - The selective sigma2-ligand Lu 28-179 has potent anxiolytic-like effects in rodents. AB - The anxiolytic potential of the selective sigma2 ligand 1-[4-[1-(4-Fluorophenyl) 1H-indol-3-yl]-1-butyl]spiro[isobenzofuran-1(3H),4-piperidine] [corrected] (Lu 28 179) was assessed in various animal models of anxiety in rodents. Lu 28-179 facilitated the exploratory behavior of mice and rats in the black and white two compartment box over a large dose range. In the rat, the minimal effective dose (MED) was 0.18 nmol/kg (0.1 microg/kg), and in the mouse, the MED was 0.00018 nmol/kg (0.1 ng/kg). The anxiolytic-like effect was maintained after treatment with 1 microg/kg/day for up to 14 days, and no anxiogenic-like effects were seen upon withdrawal from repeated treatment. Lu 28-179 increased the time that pairs of rats spent in active social interaction (unfamiliar high-light conditions), MED = 0.1 ng/kg. Daily treatment with Lu 28-179 (1.8 nmol/kg = 1 microg/kg/day) for up to 4 weeks increased the social interaction time significantly compared with controls, and no anxiogenic-like effects were seen upon withdrawal. Furthermore, Lu 28-179 reversed shock-induced suppression of drinking in the rat (MED = 18,000 nmol/kg = 10 mg/kg). Lu 28-179 did not inhibit footshock-induced ultrasonic vocalization in the rat or isolation-induced aggressive behavior in the mouse. Lu 28-179 was over 100 times more potent than diazepam in the rat and mouse black and white test box and the rat social interaction test, whereas the potency of Lu 28-179 was comparable to that of lorazepam in reversal of shock induced suppression of drinking. Lu 28-179 neither induced sedation nor impaired motor coordination, even at high doses (70,000 nmol/kg = 40 mg/kg). In conclusion, Lu 28-179 exerts potent and long-lasting anxiolytic-like effects in rodents without inducing sedation and withdrawal anxiogenesis. PMID- 9400008 TI - Neonatal administration of the selective serotonin reuptake inhibitor Lu 10-134-C increases forced swimming-induced immobility in adult rats: a putative animal model of depression? AB - Chronic administration of the tricyclic antidepressant clomipramine to neonatal rats from postnatal days 8 to 21 is reported to induce several behavioral changes in adult life, and it may serve as an animal model of human depressive disorder. Findings include increased immobility time in the forced swim test and locomotor hyperactivity in the open field test. Clomipramine is a serotonergic reuptake inhibitor, which suggests that altered development of the serotonergic system could account for the observed behavioral changes in the adult rat. The present study was carried out with a selective serotonin reuptake inhibitor (SSRI) to investigate whether the serotonin system, in particular, is involved in the neonatal animal model. The substance, Lu 10-134-C (LU), was characterized in monoamine reuptake and receptor binding assays and found to be an SSRI. Rats received LU during postnatal days 8 to 21 (2.5-15 mg/kg b. i.d.), and they were assessed in open field, forced swim and social interaction tests at the age of 4 months. Behavior of LU-treated rats and saline controls did not differ in the open field and social interaction tests. However, in the forced swim tests LU treated neonates showed prolonged immobility time compared with saline controls. In conclusion, chronic LU treatment during neonatal life produces long-term changes in the forced swim test, but not in the open field and social interaction tests. The behavioral changes in the forced swim test suggest that the central serotonergic system may be involved in the putative neonatal animal model of depression. PMID- 9400009 TI - GR89,696 is a kappa-2 opioid receptor agonist and a kappa-1 opioid receptor antagonist in the guinea pig hippocampus. AB - Receptor binding studies and electrophysiological studies demonstrated the existence of at least two kappa opioid receptors, which have been designated kappa-1 and kappa-2. Several agonists and antagonists are selective for the kappa 1 receptor whereas no known ligands are selective for the kappa-2 receptor. In this study, the kappa opioid GR89,696 was tested in the guinea pig hippocampal slice preparation for kappa-1 versus kappa-2 activity. The perforant path-evoked population spike in the dentate was use to evaluate activity at the kappa-1 receptor, and the Schaffer collateral-evoked N-methyl-D-aspartate (NMDA) receptor mediated synaptic current in CA3 pyramidal cells was used to measure kappa-2 receptor activation. GR89,696 had no effect on the perforant path-evoked dentate population spike; however, it did reverse the effects of the selective kappa-1 agonist U69,593 when co-perfused over the slices. In the CA3, GR89,696 inhibited the NMDA receptor-mediated synaptic current. The inhibition was antagonized by naloxone. The EC50 for GR89,696 on the NMDA current was 41.7 nM (95% CL, 7.0-248 nM). These findings indicate that GR89,696 is an agonist for kappa-2 opioid receptors and an antagonist at kappa-1 receptors in the guinea pig hippocampus. PMID- 9400010 TI - Role of endogenous dopamine in the neurochemical deficits induced by methcathinone. AB - Multiple administrations of methcathinone caused persistent deficits in monoaminergic systems, as reflected by decreases in dopamine and 5 hydroxytryptamine uptake capacity, tissue content and associated rate-limiting synthetic enzyme activities. Because dopamine has been implicated in mediating such effects after administration of related amphetamine analogs, its role in effecting methcathinone-induced monoaminergic neuronal impairment was assessed. A single high-dose administration of methcathinone increased striatal dopamine release, as measured by microdialysis in conscious rats and reflected by increases in striatal neurotensin-like immunoreactivity. Dopaminergic deficits observed 18 hr after a multiple-dose treatment with methcathinone were prevented by pretreatment with the selective D1 antagonist SCH23390 and D2 receptor antagonist eticlopride, but 5-hydroxytryptaminergic deficits were not altered. 5 Hydroxytryptaminergic changes did not occur in animals depleted of striatal dopamine by 6-hydroxydopamine lesions. These results indicate that dopaminergic systems are profoundly affected by methcathinone administration and that dopamine likely contributes to the monoaminergic effects of this stimulant. PMID- 9400011 TI - In vivo effects of remoxipride and aromatic ring metabolites in the rat. AB - The in vivo effects of remoxipride, in relation to some of its identified metabolites, were investigated in adult male Sprague-Dawley rats. The methods used included: (1) estimation of the in vivo rate of brain monoamine synthesis by measuring the accumulation of dihydroxyphenylalanine and 5-hydroxytryptophan after decarboxylase inhibition; (2) observations of spontaneous locomotor activity in a photocell-equipped open-field arena ( approximately 0. 5 m2); (3) treadmill locomotion ( approximately 4 m min-1); (4) inclined grid (60 degrees ) catalepsy test; (5) d-amphetamine-induced (1.0 mg kg-1) hyperlocomotion;(6) quinpirole-induced (0.4 mg kg-1) hypothermia. By use of one or more of these tests, the findings with remoxipride were as follows: First, remoxipride had a late onset of action (up to 3 h). Second, potency and efficacy depended on exposure to hepatic metabolism. Thus, intraperitoneal administration was more effective than the subcutaneous route, whereas virtually all biological effects were lost on intracerebroventricular administration. The ED50 values (micromol kg 1, neostriatal dihydroxyphenylalanine accumulation) for remoxipride and a range of its phenolic aromatic ring metabolites were: remoxipride (approximately 20), NCQ-344 (approximately 0.01), FLA-797 (approximately 0.1), FLA-908 (approximately 2.2), NCQ-436 (approximately 25) and NCQ-469 (approximately 30). Considering remoxipride as a nonclozapine atypical antipsychotic drug, together with the fact that remoxipride behaves as a prodrug in the laboratory studies above, further characterization of the pharmacodynamic profile of its metabolites remains a challenge. PMID- 9400012 TI - Involvement of alpha-2 adrenoceptors in the effects of moxonidine on intestinal motility and fluid transport. AB - The aims of this study were to examine how the imidazoline (I)1/alpha-2 receptor agonist moxonidine and the putative endogenous imidazoline receptor agonist agmatine might affect intestinal motility and fluid transport. The effects of moxonidine were compared with those of UK 14,304, a highly selective alpha-2 adrenoceptor agonist with very low affinity for I1 receptors. Moxonidine and UK 14,304 inhibited the peristaltic reflex in the isolated rat ileum. The inhibitory effects were antagonized by the selective alpha-2 adrenoceptor antagonist yohimbine and the I1/alpha-2 antagonist efaroxan and almost completely blocked by the irreversible alpha-2 adrenoceptor antagonist EEDQ (N-ethoxycarbonyl-2-ethoxy 1,2-dihydroquinoline), which has a low affinity for imidazoline receptors. Yohimbine (3 microM) and efaroxan (0.01 and 1 microM) caused parallel rightward shifts to the concentration-response curves of moxonidine and UK 14,304, yielding pKB values corresponding to those at alpha-2 binding sites. Moxonidine induced dose-dependent proabsorptive effects in the jejunum and ileum and also reversed the secretory phase of the vasoactive intestinal peptide-induced responses. The degree of antagonism by yohimbine and efaroxan was similar against moxonidine and UK 14,304 on the proabsorptive and antisecretory effects. We conclude that the effects of moxonidine in mediating inhibition of intestinal motility and enhancing fluid transport are attributed predominantly to interaction with alpha 2 adrenoceptors. Agmatine had no effect on peristalsis but significantly decreased the rate of fluid absorption from the jejunum and ileum, an effect in contrast to moxonidine. A physiological role for agmatine in the regulation of intestinal transport remains to be clarified. PMID- 9400013 TI - Neutrophil cytotoxicity of the chemically reactive metabolite(s) of clozapine: possible role in agranulocytosis. AB - Clozapine is associated with a 0.8% incidence of agranulocytosis. Bioactivation to an unstable protein-reactive metabolite, identified as a nitrenium intermediate, has been implicated in the toxicity. In this study, we investigated whether the reactive metabolite is cytotoxic toward polymorphonuclear leukocytes and mononuclear leukocytes using horseradish peroxidase and H2O2 to generate the metabolite in situ. In the absence of a full metabolizing system (i. e., lack of horseradish peroxidase and/or H2O2), clozapine (0-100 microM) and its stable metabolites were not cytotoxic. With a full metabolizing system, both clozapine (30 microM) and demethylclozapine exhibited cytotoxicity toward polymorphonuclear leukocytes (50.7 +/- 7.7% and 17.6 +/- 1.2% cell death, respectively) and mononuclear leukocytes (36.6 +/- 2.1% and 24.6 +/- 4.1%, respectively), whereas clozapine N-oxide was not cytotoxic. Exogenous glutathione (GSH), N acetylcysteine and ascorbic acid all protected the cells. Bioactivation of clozapine and demethylclozapine, but not the N-oxide, was accompanied by depletion of intracellular GSH. [14C]Clozapine was metabolized to the previously identified C6 and C9 glutathionyl conjugates; GSH conjugates were also detected when demethylclozapine and clozapine N-oxide were bioactivated by horseradish peroxidase and H2O2. In conclusion, using a novel in vitro assay, we have shown that clozapine and its stable metabolites are not cytotoxic per se but are bioactivated to cytotoxic metabolites. The cytotoxic metabolite of clozapine is identical to the protein-reactive metabolite that has been characterized previously. These cytotoxic metabolites may play an important role in the pathogenesis of clozapine agranulocytosis; the mechanism by which this occurs is currently being investigated. PMID- 9400014 TI - Cell cycle-dependent chronotoxicity of irinotecan hydrochloride in mice. AB - The mechanisms underlying the circadian rhythm of the toxicity induced by irinotecan hydrochloride (CPT-11; 7-ethyl-10-[4-(1-piperidino)-1 piperidino]carbonyloxycamptothecin) were investigated from the viewpoint of the sensitivity of living organisms and the pharmacokinetics of the drug. ICR male mice were housed under standardized light-dark cycle conditions (lights on at 0700, off at 1900) with food and water ad libitum. The loss of body weight after an intraperitoneal injection of CPT-11 (100 mg/kg) was more serious in the late dark and the early light and milder in the late light and the early dark. The CPT 11-induced leukopenia was more serious in the late dark and milder in the late light. The lower toxicity of CPT-11 was observed when DNA synthesis and type I DNA topoisomerase activity in bone marrow cells decreased and the higher toxicity was observed when these activities began to increase. There were circadian stage dependent changes in the concentrations of CPT-11 and its major metabolite (SN 38; 7-ethyl-10-hydroxycamptothecin) in plasma. The higher concentrations of CPT 11 and SN-38 in plasma were observed when the level of CPT-11-induced toxicity increased. The present study suggests that the toxicity of CPT-11 is influenced by circadian rhythm-dependent processes. PMID- 9400015 TI - Abnormality in plasma catecholamines and myocardial adrenoceptors in cardiomyopathic BIO 53.58 Syrian hamsters and improvement by metoprolol treatment. AB - The catecholaminergic neuronal activity and the densities of alpha-1 and beta adrenoceptors and angiotensin II receptors were simultaneously determined in BIO 53.58, a model of idiopathic dilated cardiomyopathy, and F1B control hamsters. Further, we examined the effect of repeated p.o. administration of metoprolol on these biochemical parameters. Compared with F1B control hamsters, there was a significant decrease in Bmax of specific binding of both (-) [125I]iodocyanopindolol and [3H]prazosin with a marked elevation of plasma catecholamine (mainly norepinephrine and epinephrine) concentrations, in BIO 53.58 hamsters at 11 and 18 weeks of age (severe cardiomyopathic stage), but not at 5 weeks of age. On the other hand, the Bmax value of myocardial [125I]angiotensin II binding in BIO 53.58 hamsters was almost identical to that in F1B hamsters. These results suggest a development of down-regulation of myocardial beta and alpha-1 adrenoceptors because of an increased catecholaminergic neuronal activity with aging in BIO 53.58 hamsters. Repeated p.o. administration of a relatively low dose (1 mg/kg/day) of metoprolol for 7 weeks in 11-week-old BIO 53.58 hamsters caused a significant increase of myocardial (-)-[125I]iodocyanopindolol binding sites with a marked reduction in plasma catecholamine levels; this indicated a significant recovery to the F1B levels. The improvement of these biochemical parameters by metoprolol treatment was also accompanied by a significant decrease in the fibrosis in the heart in BIO 53.58 hamsters. These data suggest that catecholaminergic neurons and adrenoceptors play a part in the development of heart failure in idiopathic dilated cardiomyopathy. Consequently, the present study may provide a further pharmacological basis for the use of beta-1 adrenoceptor antagonists in patients with idiopathic dilated cardiomyopathy. PMID- 9400016 TI - Neuronal nicotinic acetylcholine receptor activation modulates gamma-aminobutyric acid release from CA1 neurons of rat hippocampal slices. AB - In the present study we investigated electrophysiologically the nicotinic responses of pyramidal neurons and interneurons visualized by infrared-assisted videomicroscopy and fluorescence in the CA1 field of hippocampal slices obtained from 8- to 24-day-old rats. Application of nicotinic agonists to CA1 neurons evoked at least four types of nicotinic responses. Of major interest was the ability of these agonists to induce the release of gamma-aminobutyric acid (GABA) from interneurons. Slowly decaying ACh whole-cell currents and GABA-mediated postsynaptic currents could be recorded from pyramidal neurons and interneurons, whereas fast-decaying nicotinic currents and fast current transients were recorded only from interneurons. Nicotinic responses were sensitive to blockade by d-tubocurarine (10 microM), which indicated that they were mediated by nicotinic acetylcholine receptors (nAChRs). The slowly decaying currents, the postsynaptic currents and the fast current transients were insensitive to blockade by the alpha-7 nAChR-specific antagonist methyllycaconitine (up to 1 microM) or alpha-bungarotoxin (100 nM). On the other hand, the slowly decaying nicotinic currents recorded from the interneurons were blocked by the alpha4beta2 nAChR-specific antagonist dihydro-beta-erythroidine, and the fast-desensitizing nicotinic currents were evoked by the alpha-7 nAChR-specific agonist choline. In experimental conditions similar to those used to record nicotinic responses from neurons in slice (i. e., in the absence of tetrodotoxin), we observed that nicotinic agonists can also induce the release of GABA from hippocampal neurons in culture. In summary, these results provide direct evidence for more than one subtype of functional nAChR in CA1 neurons and suggest that activation of nAChRs present in GABAergic interneurons can evoke inhibitory activity in CA1 pyramidal neurons, thereby modulating processing of information in the hippocampus. PMID- 9400017 TI - -(S)-Alpha-phenyl-2-pyridine-ethanamine Dihydrochloride-, a low affinity uncompetitive N-methyl-D-aspartic acid antagonist, is effective in rodent models of global and focal ischemia. AB - [(S)-Alpha-phenyl-2-pyridine-ethanamine dihydrochloride] (ARL 15896AR) is a low affinity uncompetitive N-methyl-D-aspartic acid receptor antagonist that was tested in animal models of anoxia and ischemia. Pretreatment of rodents with ARL 15896AR extended survival time during exposure to hypoxia. With the rat four vessel occlusion model of global ischemia (20 min), oral dosing commencing at reflow, resulted in significant protection of the CA1 hippocampal neurons. ARL 15896AR was, however, ineffective in the rat two-vessel occlusion model and in the gerbil models of forebrain ischemia, the latter due to an inability to attain suitable plasma levels. In the spontaneously hypertensive rat model of middle cerebral artery occlusion (MCAO) (2 hr plus 22 hr reflow), acute dosing with ARL 15896AR (i.p.) beginning from 30 min before or up to 1 hr post-MCAO significantly reduced cortical infarct volume. The ability of ARL 15896AR to influence infarct size, as well as functional correlates was examined in SHR after 90 min of MCAO. T2 weighted magnetic resonance images taken at 2 and 6 days post-MCAO revealed significantly smaller lesion sizes in the group receiving injections with ARL 15896AR beginning 30 min after occlusion. Spontaneously hypertensive rats were subsequently tested (30-42 days post-MCAO) and found to be deficient in skilled use of the forepaws (staircase test). The contralateral forepaw was most severely impaired, however, ARL 15896AR treatment prevented motor impairment in only the ipsilateral forepaw. Histopathological examination of cortical infarct size was unremarkable between treated and control rats. The findings indicate that ARL 15896AR exhibits neuroprotection in global and focal models of ischemia PMID- 9400018 TI - Isolation, heterologous expression and functional characterization of a novel cytochrome P450 3A enzyme from a canine liver cDNA library. AB - A cDNA encoding a new member of the cytochrome P450 3A subfamily, P450 3A26, has been isolated from phenobarbital-induced canine liver. The sequence encodes a protein of 503 amino acids with 33 nucleotide differences conferring 22 amino acid substitutions when compared with the previously identified canine CYP3A12 enzyme. Nine of the amino acid differences are within the substrate recognition sites (SRSs) identified for P450 family 2, with five residue substitutions clustered within SRS-6. To facilitate heterologous expression in Escherichia coli, the N-terminus of 3A26 was modified. The expressed protein comigrated with a 3A-immunoreactive protein in dog liver microsomes with a slightly greater electrophoretic mobility on sodium dodecyl sulfate-polyacrylamide gel electrophoresis than 3A12, which suggests that 3A26 corresponds to a previously noted but never characterized 3A enzyme in dogs. Functional characterization of 3A26 was undertaken with use of progesterone, testosterone and androstenedione as substrates. Assays of expressed 3A26 and 3A12 demonstrated that 3A26 displays low steroid hydroxylase activity. Identification of an additional canine 3A enzyme should increase our understanding of xenobiotic metabolism in this important animal model. These findings also suggest that 3A26 and 3A12 may be an interesting model system for the investigation of structure-function relationships involved in steroid metabolism catalyzed by members of the cytochrome P450 3A subfamily. PMID- 9400019 TI - In vitro pharmacological characterization of PD 166285, a new nanomolar potent and broadly active protein tyrosine kinase inhibitor. AB - PD 166285, a novel protein tyrosine kinase inhibitor of a new structural class, the 6-aryl-pyrido[2,3-d]pyrimidines, was synthesized as the most potent and soluble analog of a series of small molecules originally identified by screening a compound library with assays that measured protein tyrosine kinase activity. PD 166285 was found to inhibit Src nonreceptor tyrosine kinase, fibroblast growth factor receptor-1, epidermal growth factor receptor and platelet-derived growth factor receptor beta subunit (PDGFR-beta), tyrosine kinases with half-maximal inhibitory potencies (IC50 values) of 8.4 +/- 2.3 nM (n = 6), 39.3 +/- 2.8 nM (n = 16), 87.5 +/- 13.7 nM (n = 6) and 98.3 +/- 7.9 nM (n = 16), respectively. PD 166285 also demonstrated inhibitory activity against mitogen-activated protein kinase (IC50 = 5 microM) and protein kinase C (IC50 = 22.7 microM). PD 166285 was further characterized as an ATP competitive inhibitor of Src nonreceptor tyrosine kinase, PDGFR-beta, fibroblast growth factor receptor-1 and epidermal growth factor receptor tyrosine kinases. In addition, PD 166285 inhibited PDGF- and EGF stimulated receptor autophosphorylation in vascular smooth muscle cells (VSMCs) and A431 cells, respectively, and basic fibroblast growth factor-mediated tyrosine phosphorylation in Sf9 cells, with IC50 values of 6.5 nM, 1.6 microM and 97.3 nM, respectively, further establishing a tyrosine kinase mechanism of inhibition. The inhibition of PDGF receptor autophosphorylation in VSMCs by PD 166285 was long lasting and persisted for 4 days after a single 1-hr exposure followed by extensive washing. The PDGF-induced tyrosine phosphorylation of the 44- and 42-kDa mitogen-activated protein kinase isoforms was also blocked as a result of the inhibition of PDGF-stimulated receptor autophosphorylation by PD 166285 in VSMCs. The effects of PD 166285 were also demonstrated in functional assays of cell attachment, migration and proliferation, in which vascular cell adhesion to vitronectin, PDGF-directed chemotaxis and serum-stimulated cell growth were all potently inhibited with IC50 values of 80 yo 120 nM. Finally, PD 166285 uniquely demonstrated potent inhibition of phorbol ester-induced production of 92-kDa gelatinase A (MMP-9) in VSMC without affecting 72-kDa gelatinase B (MMP-2) as measured by gelatin zymography. These results highlight the biological characteristics of PD 166285 as a broadly active protein tyrosine kinase capable of potently inhibiting a number of kinase mediated cellular functions, including cell attachment, movement and replication. The potential therapeutic utility of this broadly acting inhibitor as an antiproliferative and antimigratory agent could extend to such diseases as cancer, atherosclerosis and restenosis, in which redundancies in protein kinase signaling pathways are known to exist. PMID- 9400020 TI - Enhanced dopamine release and phosphorylation of synapsin I and neuromodulin in striatal synaptosomes after repeated amphetamine. AB - Repeated, intermittent treatment of rats with amphetamine followed by a withdrawal period leads to an enhancement in amphetamine-induced dopamine release. We previously reported an increased stoichiometry of site 3-phospho synapsin I and increased levels of phospho-Ser41-neuromodulin in striatum after repeated amphetamine. In this study, we examined whether the enhanced amphetamine induced dopamine release and increased levels of these phosphoproteins would be detected in synaptosomes from rats pretreated and withdrawn from repeated amphetamine. Enhanced amphetamine-induced dopamine release was detected in striatal synaptosomes from rats treated with repeated amphetamine compared with controls. The enhanced dopamine release was Ca++ dependent. State-specific antibodies were used to measure the levels of site 3-phospho-synapsin I, phosphorylated by CaM kinase II, and phospho-Ser41-neuromodulin, phosphorylated by protein kinase C, in incubated striatal S1 fractions and synaptosomes. The levels of site 3-phospho-synapsin I and phospho-Ser41-neuromodulin were increased by 40% and 30%, respectively, in amphetamine-pretreated rats compared with controls. Total neuromodulin and synapsin I was not altered. There was a significant 26% increase in CaM kinase II activity in the synaptosomes from amphetamine-pretreated rats but no change in content. No change in protein kinase C activity or content of the alpha-isozyme was detected after repeated amphetamine. Our results demonstrate that the enhanced amphetamine-induced dopamine release and occurring after repeated amphetamine can be detected in synaptosome preparations. Repeated amphetamine leads to alterations in phosphorylation/dephosphorylation activities that can be detected in the incubated synaptosomes. Because the enhanced amphetamine-induced dopamine release after repeated amphetamine appears to be Ca++ sensitive, it is possible that the altered phosphorylation systems, and perhaps site 3-phospho-synapsin I and phospho-Ser41-neuromodulin, play a role in the enhanced dopamine release. PMID- 9400021 TI - Attenuation by phosphodiesterase inhibitors of lipopolysaccharide-induced thromboxane release and bronchoconstriction in rat lungs. AB - Exposure of perfused rat lungs to lipopolysaccharides (LPS) causes induction of cyclooxygenase-2 followed by thromboxane (TX)-mediated bronchoconstriction (BC). Recently, phosphodiesterase (PDE) inhibitors have received much interest because they not only are bronchodilators but also can suppress release of proinflammatory mediators. In the present study, we investigated the effect of three different PDE inhibitors on TX release and BC in LPS-exposed perfused rat lungs. The PDE inhibitors used were motapizone (PDE III specific), rolipram (PDE IV specific), and zardaverine (mixed PDE III and IV specific). At 5 microM, a concentration at which all three compounds selectively block their respective PDE isoenzyme, rolipram (IC50 = 0.04 microM) and zardaverine (IC50 = 1.8 microM) largely attenuated the LPS-induced BC, whereas motapizone was almost ineffective (IC50 = 40 microM). In contrast to LPS, BC induced by the TX-mimetic U46619 was prevented with comparable strength by motapizone and rolipram. In LPS-treated lungs, the TX release was reduced to 50% of controls by rolipram and zardaverine but was unaltered in the presence of 5 microM motapizone. Increasing intracellular cAMP through perfusion of db-cAMP or forskolin (activates adenylate cyclase) also reduced TX release and BC. We conclude that PDE inhibitors act via elevation of intracellular cAMP. Although both PDE III and PDE IV inhibitors can relax airway smooth muscle, in the model of LPS-induced BC, PDE IV inhibitors are more effective because (in contrast to PDE III inhibitors) they also attenuate TX release. PMID- 9400022 TI - Ochratoxin A-induced stimulation of extracellular signal-regulated kinases 1/2 is associated with Madin-Darby canine kidney-C7 cell dedifferentiation. AB - The kidneys represent one of the main targets of ochratoxin A (OTA), a secondary fungal metabolite that is produced by certain species of Aspergillus and Penicillium. OTA has the ability to disturb Madin-Darby canine kidney (MDCK) cell pH homeostasis, leading to intracellular alkalinization and morphological alterations resembling those that occur when MDCK cells are exposed to transient alkaline stress. Because alkali-induced epithelial dedifferentiation of MDCK-C7 cells is associated with an increase in the activity of extracellular signal regulated kinases (ERK), we performed experiments that investigated a possible role for ERK1 and ERK2 as intracellular signaling molecules mediating some of the mycotoxin's effects on renal epithelia. We studied the effects of OTA on ERK1/2 phosphorylation and activation, as well as on cell morphology by using cloned MDCK-C7 and MDCK-C11 cells. In MDCK-C7 cells, but not in MDCK-C11 cells, OTA led to a time-dependent and concentration-dependent increase in ERK1/2 phosphorylation. OTA-induced ERK1/2 phosphorylation in MDCK-C7 cells occurred at concentrations of 500 nM, started after 2 hr and was maximal after 8 hr. Furthermore, after 8 hr of incubation, 500 nM and 1 microM OTA significantly increased ERK1/2 activity in MDCK-C7 but not in MDCK-C11 cells. This OTA stimulated ERK1/2 phosphorylation and ERK1/2 activation in MDCK-C7 cells was partially inhibited by the synthetic mitogen-activated protein kinase kinase (MKK or MEK) inhibitor PD098059. Transepithelial resistance and lactate dehydrogenase release remained unaltered after incubation in the presence of 1 microM OTA for 8 hr or of 100 nM OTA for 24 hr, so it is unlikely that these OTA effects on ERK1/2 are due to secondary toxic effects of the mycotoxin. Interestingly, OTA-induced long-term activation of ERK1/2 in MDCK-C7 cells was associated with epithelial dedifferentiation, as assessed by analysis of vectorial solute and water transport as well as cell morphology. In contrast, MDCK-C11 cells, which do not show significant increases in ERK1/2 phosphorylation and ERK1/2 activity in response to OTA, retained their epithelial phenotype under identical experimental conditions. Taken together, our data demonstrate an epithelial dedifferentiation of MDCK-C7 cells, but not of MDCK-C11 cells, after long-term incubation in the presence of OTA, a result associated with the ability of this mycotoxin to stimulate ERK1/2 in MDCK-C7 cells but not in MDCK-C11 cells. We conclude that OTA induced activation of ERK1/2 could be an important intracellular signaling pathway that mediates some of the mycotoxin's effects on renal epithelia. PMID- 9400023 TI - Influence of streptozotocin diabetes on the alpha-1 adrenoceptor and associated G proteins in rat arteries. AB - Previous studies from this laboratory have demonstrated an enhancement in both the contractile and signaling response to stimulation of either alpha-1 adrenoceptors or guanine nucleotide binding proteins (G proteins) in arteries from male Wistar rats with 12 to 14 weeks of streptozotocin-induced diabetes. The purpose of the present investigation was to determine whether changes in arterial alpha-1 adrenoceptors or the G proteins coupled to them are associated with the enhanced responsiveness of the diabetic arteries. No difference in affinity was detected between control and diabetic aorta or caudal artery membranes in saturation binding of [3H]prazosin to alpha-1 adrenoceptors. However, the alpha-1 adrenoceptor number was significantly decreased in caudal artery but not aorta from diabetic rats. In competition binding experiments, a low-affinity and a high affinity binding site for norepinephrine were detected in the absence of guanine nucleotides and NaCl in control arteries, whereas only the low-affinity site was detected in diabetic arteries, suggesting that coupling of the alpha-1 adrenoceptor to G proteins is impaired in diabetic aorta and caudal artery. The levels of immunoreactive Gi2,3alpha and Gq/11alpha were not different between control and diabetic aorta or caudal artery. Thus, not only do changes in the number and coupling of the alpha-1 adrenoceptor or level of G proteins not explain the enhanced contractile responses of diabetic arteries to norepinephrine, but also the changes in alpha-1 adrenoceptor binding would counteract the enhancement. Instead, an increase in the activity of the G proteins or phospholipase C-beta coupled to the alpha-1 adrenoceptor may be mediating the enhanced responsiveness elicited by alpha-1 adrenoceptor stimulation in diabetic arteries. PMID- 9400024 TI - Irreversible inhibition of cytochrome P450 by nitric oxide. AB - Nitric oxide (NO) modulates various metabolisms through interaction with thiol proteins and hemoproteins. Although NO interacts reversibly with iron moieties of heme proteins, including cytochrome P450 (P450), dynamic aspects of the formation, catalytic functions and fates of NO-P450 adducts remain to be elucidated. When incubated with NOC7, which spontaneously and stoichiometrically releases NO within 5 min, microsomal P450 rapidly formed nitrosyl-heme adducts as determined by the electron spin resonance method. The signal intensity for the complex increased with time, peaking at 30 min and decreasing to below detectable levels by 60 min of incubation. In contrast, the microsomal levels of low-spin ferric forms of P450 (g = 2.26) rapidly decreased during the initial 30 min but recovered time-dependently thereafter. Analysis by differential spectra (reduced form/CO-reduced form) revealed that on incubation with NOC7, the form of microsomal P450 also changed in a biphasic manner. To elucidate the mechanism for the decrease in the levels of P450, microsomal levels of P450 isozymes (CYPs) were determined by Western blot analysis using specific antibodies against CYP3A2 and CYP2C11, major isoforms found in male rat liver. Kinetic analysis revealed that no appreciable degradation of P450 proteins occurred during the incubation of microsomes with NOC7. The effect of NO on the catalytic activity of the enzymes was determined by using testosterone as substrate because hydroxylation of steroid hormones is one of the major functions of P450. When exposed to NO, the hydroxylation activity in microsomes rapidly decreased during the initial 10 min and then disappeared slowly. These results suggested that NO formed dissociable complexes with P450 isozymes and the catalytic functions of these isozymes were irreversibly inactivated after dissociation from their heme moiety. PMID- 9400025 TI - Hormonal regulation of microsomal cytochrome P450 2C11 in rat liver and kidney. AB - The current study was conducted to investigate hormonal regulation of cytochrome P450 2C11 (CYP2C11) in rat liver and kidney of adult male rats. In two experiments, hypophysectomy (Hx) resulted in decreased (P < .05) hepatic CYP2C11 apoprotein and mRNA levels. Growth hormone (GH) replacement of Hx rats prevented the decline in hepatic CYP2C11 apoprotein and mRNA levels, whereas, subcutaneous injection of testosterone had no effect. Rat pituitary extract is equally effective in intact or castrated Hx rats in preventing the decline in hepatic CYP2C11 apoprotein and mRNA levels. Specific neutralization of rat GH by sheep anti-rat GH serum reduced (P < . 05) serum IGF-I concentrations, hepatic CYP2C11 apoprotein and mRNA levels. Hx of male rat resulted in decreased (P < .05) renal CYP2C11 apoprotein and mRNA levels, and treatment with GH failed to prevent these effects; however, supplementation of Hx rats with testosterone or rat pituitary extract prevented the Hx-induced decrease of renal CYP2C11 apoprotein and mRNA levels, and the effects of rat pituitary extract occurred only in intact rats. Neutralization of rat GH by anti-rGH significantly reduced (P < .05) CYP2C11 mRNA levels and serum T concentrations but not serum LH concentrations. These results indicate that although hepatic CYP2C11 is regulated by GH, rat renal CYP2C11 is regulated primarily by gonadal steroids. PMID- 9400026 TI - Possible involvement of 5-HT2 receptor activation in aggravation of diet-induced acute pancreatitis in mice. AB - Acute pancreatitis was induced in mice by feeding with a choline-deficient ethionine-supplemented diet. All the mice developed acute pancreatitis, and approximately 80% of them died within 4 days. Stereomicroscopic and light microscopic examinations revealed that pancreatic necrosis and circulatory disturbance that were not apparent on day 1 were increased markedly on days 2 and 3. Serum levels of pancreatic enzymes were normal or reduced on day 1 but then increased to peak on day 3. Plasma 5-hydroxyindoleacetic acid levels, which may indicate serotonin release, were significantly increased on days 1 through 3. Pretreatment with D, L-p-chlorophenylalanine methylester hydrochloride (200-400 mg/kg) significantly attenuated the mortality of the mice with pancreatitis. Dose dependent attenuation was also obtained with ketaserin (0. 01-10 mg/kg), cyproheptadine (0.01-10 mg/kg), pindolol (0.1-100 mg/kg) and NAN-190 (0.1-100 mg/kg), but not with 0.01 to 10 mg/kg of ICS205-930 or M-840, and the activities were significantly correlated with the binding affinities for serotonin2 receptor on the rat cerebral cortex. In addition, ketanserin or cyproheptadine attenuated the morphologic changes in the choline-deficient ethionine-supplemented diet mice at a dose (3.2 mg/kg) that hardly affected the serum enzyme levels. We propose that serotonin2 receptor activation plays an important role in the aggravation of diet-induced acute pancreatitis. PMID- 9400027 TI - Chronic administration of a glycine partial agonist alters the expression of N methyl-D-aspartate receptor subunit mRNAs. AB - Both acute and chronic treatments with the glycine partial agonist 1 aminocyclopropanecarboxylic acid (ACPC) are neuroprotective in animal models of focal, global and spinal ischemia. After a chronic regimen of ACPC, brain and plasma levels were undetectable at the time of ischemic insult, which suggests that the neuroprotective effects of acute and chronic ACPC are mediated by different mechanisms. To investigate the possibility that chronic administration of ACPC alters N-methyl-D-aspartate (NMDA) receptor composition, the levels of mRNAs encoding zeta and epsilon subunits were quantified by in situ hybridization histochemistry with 35S-labeled riboprobes. Chronic ACPC administered to mice (200 mg/kg for 14 days) increased the level of epsilon-1 mRNA in the hippocampus (particularly CA1 and CA2 regions) and cerebral cortex (frontal, parietal and occipital regions), without altering levels in cerebellum. In contrast, this regimen decreased epsilon-3 subunit mRNA levels in the hippocampus (especially CA1 and dentate gyrus) and frontal and occipital cortices. Decreases in epsilon-2 subunit mRNA levels in cerebral cortex (especially frontal and parietal cortices) were also observed without accompanying alterations in the cerebellum, hippocampus or dentate gyrus. The levels of zeta subunit mRNA (determined with a probe that detects all splice variants) were not altered in any brain areas examined. Based on studies in recombinant receptors, these region-specific changes in mRNAs produced by a chronic regimen of ACPC could result in NMDA receptors with reduced affinities for glycine and glutamate. It is hypothesized that such alterations in NMDA receptor subunit composition may explain the neuroprotective effects produced by chronic ACPC. PMID- 9400028 TI - Discovery of less nephrotoxic FK506 analogs and determining immunophilin dependence of immunosuppressant nephrotoxicity with a novel single-dose rat cisplatin potentiation assay. AB - Comparing nephrotoxicity of numerous drug analogs is impractical with chronic in vivo models. We devised a new cisplatin potentiation assay (CISPA) that sensitively detects renal injury as a serum creatinine increase when only one dose of test compound is followed by cisplatin. Reference nephrotoxins known to act on various sites in kidney tubules, glomeruli or renal papilla were all detected by the CISPA at single doses that without cisplatin gave little change, which showed that this simple, sensitive assay has broad potential utility for mechanistic studies of nephrotoxicity. We used the CISPA both to probe the nephrotoxic mode of action of immunosuppressants and to search for safer compounds. Although several non-nephrotoxic immunosuppressants were inactive, cyclosporine, FK506, ascomycin (C21-ethyl-FK506) and rapamycin were nephrotoxic in the CISPA at single doses equal to the daily amounts required to reduce creatinine clearance with 14 days of treatment. Similar therapeutic indices were derived comparing toxicity by either method to prevention of rat ear-heart allograft rejection. C18-OH-ascomycin, an FK506-binding protein (FKBP) antagonist, reversed in vivo immunosuppression by FK506 and ascomycin in the rat, and pretreatment in the CISPA blocked FK506 and ascomycin nephrotoxicity, which showed a common immunophilin dependence. Rapamycin nephrotoxicity was unaffected (as with cyclosporine), which indicated that binding to FKBP was not required. Rapamycin nephrotoxicity thus appears mechanistically unrelated to its immunosuppressive mode of action. Screening with the CISPA enabled discovery of A 119435, a less nephrotoxic ascomycin analog having a 10-fold higher therapeutic index. PMID- 9400029 TI - The protective effect of metallothionein against lipid peroxidation caused by retinoic acid in human breast cancer cells. AB - The treatment of breast cancer by retinoic acid (RA) may be mediated by lipid peroxidation. Expression of metallothionein (MT) in cancer cells, however, can protect against lipid peroxidation by scavenging hydroxyl radicals. In this study, a two-by-six factorial design was used to investigate the interactive effects of all-trans-RA and zinc (Zn)-induced MT on the growth of two human breast cancer cell lines differing in basal expression of MT and estrogen receptors; MCF7 cells express estrogen receptor, BT-20 cells do not. Cells were treated with Zn to induce MT and then treated with six RA concentrations. Cell proliferation, lipid peroxidation, MT protein, MT mRNA and glutathione concentrations were measured. BT-20 cells expressed higher constitutive MT concentrations than MCF7 cells. MT was significantly increased by Zn treatment in BT-20 cells but not in MCF7 cells. Low RA concentrations stimulated growth proliferation but higher concentrations inhibited cell proliferation. Elevated RA concentrations increased lipid peroxidation as measured by thiobarbituric acid reactive substances. There was a significant negative correlation between lipid peroxidation and cell proliferation. Growth inhibition and lipid peroxidation were reduced by Zn pretreatment in BT-20 cells but not in MCF7 cells. RA increased MT levels in both cell lines, which suggests that RA may generate free radicals which will induce MT mRNA expression. Glutathione did not appear to be a significant factor. Therefore, induction of MT by Zn may modulate the growth inhibitory effects of RA in human breast cancer cells. One mechanism of growth inhibition may be through increased lipid peroxidation. Induction of MT by RA may be one explanation for acquired RA resistance in cancer. PMID- 9400030 TI - Enhanced renal toxicity by inorganic mercury in metallothionein-null mice. AB - To elucidate a protective role of metallothionein (MT) in the manifestation of inorganic mercury toxicity, we studied the susceptibility of MT-null mice to the renal toxicity of mercuric chloride. Because the MT-null (J) mice are a genetic background of 129/Sv strain, the 129/Sv mice were used as wild-type controls. Nine-week-old male MT-null (J) and 129/Sv mice were given subcutaneous injections of mercuric chloride at doses of 10 to 40 micromol/kg. The basal MT level in the kidney of MT-null (J) mice was undetectable (<0.2 microg/g of tissue) and approximately 2.5 microg/g of tissue in 129/Sv mice. The sensitivity to the renal toxicity of mercuric chloride was markedly enhanced in the MT-null (J) mice compared with the 129/Sv mice. The renal mercury level was similar for the MT null (J) and 129/Sv mice at 4 hr after the injection of mercuric chloride (20 micromol/kg) but became significantly lower in MT-null (J) mice than in 129/Sv mice at 24 and 72 hr. Based on the present results, we conclude that MT is an important protective factor against the renal toxicity caused by inorganic mercury and that it may play a major role in the retention of mercury in the kidney. PMID- 9400031 TI - Novel qualitative structure-activity relationships for the antinociceptive actions of H2 antagonists, H3 antagonists and derivatives. AB - Recent studies have shown that cimetidine, burimamide and improgan (also known as SKF92374, a cimetidine congener lacking H2 antagonist activity) induce antinociception after intracerebroventricular administration in rodents. Because these substances closely resemble the structure of histamine (a known mediator of some endogenous analgesic responses), yet no role for known histamine receptors has been found in the analgesic actions of these agents, the structure-activity relationships for the antinociceptive effects of 21 compounds chemically related to H2 and H3 antagonists were investigated in this study. Antinociceptive activity was assessed on the hot-plate and tail-flick tests after intracerebroventricular administration in rats. Eleven compounds induced time dependent (10-min peak) and dose-dependent antinociceptive activity with no observable behavioral impairment. ED50 values, estimated by nonlinear regression, were highly correlated across nociceptive assays (r2 = 0.98, n = 11). Antinociceptive potencies varied more than 6-fold (80-464 nmol), but were not correlated with activity on H1, H2 or H3 receptors. Although highly potent H3 antagonists such as thioperamide lacked antinociceptive activity, homologs of burimamide and thioperamide containing N-aromatic substituents retained H3 antagonist activity and also showed potent, effective analgesia. A literature review of the pharmacology of these agents did not find a basis for their antinociceptive effects. Taken with previous findings, the present results suggest: 1) these compounds act on the brain to activate powerful analgesic responses that are independent of known histamine receptors, 2) the structure activity profile of these agents is novel and 3) brain-penetrating derivatives of these compounds could be clinically useful analgesics. PMID- 9400032 TI - Nephrotoxicity of the glutathione and cysteine S-conjugates of the sevoflurane degradation product 2-(fluoromethoxy)-1,1,3,3, 3-pentafluoro-1-propene (Compound A) in male Fischer 344 rats. AB - 2-(Fluoromethoxy)-1,1,3,3,3-pentafluoro-1-propene (Compound A) is a halogenated alkene that is nephrotoxic in rats when administered by inhalation or intraperitoneally. Compound A undergoes glutathione-dependent metabolism: Compound A-derived glutathione S-conjugates and mercapturates are excreted in the bile and urine, respectively, of rats given Compound A. The present experiments were designed to study the nephrotoxicity of the Compound A-derived glutathione and cysteine S-conjugates, S-[2-(fluoromethoxy)-1,1,3,3, 3 pentafluoropropyl]glutathione , S-[2-(fluoromethoxy)-1,3,3, 3-tetrafluoro-1 propenyl]glutathione , S-[2-(fluoromethoxy)-1,1,3,3, 3-pentafluoropropyl]-L cysteine and S-[2-(fluoromethoxy)-1,3,3, 3-tetrafluoro-1-propenyl]-L-cysteine . Conjugates , and given intraperitoneally produced dose-dependent nephrotoxicity that was characterized by diuresis, increased excretion of glucose and protein, elevated blood urea nitrogen concentrations and severe morphological changes in the kidneys, particularly in the proximal tubules. Glutathione S-conjugate , at a dose of 500 micromol/kg, was hepatotoxic. Cysteine S-conjugate was not nephrotoxic, apparently because of its facile cyclization to the thiazoline 2-[1 (fluoromethoxy)-2,2,2-trifluoroethyl]-4,5-dihydro-1, 3-thiazole-4-carboxylic acid, which is not a beta-lyase substrate. Also, the alpha-methyl analog of cysteine S-conjugate S-[2-(fluoromethoxy)-1,1,3,3, 3-pentafluoropropyl]-DL-alpha methylcysteine, which cannot undergo beta-lyase-dependent bioactivation, was not nephrotoxic. These in vivo data show that Compound A-derived S-conjugates are nephrotoxic and that the toxicity is associated with beta-lyase-dependent bioactivation. PMID- 9400033 TI - Characterization of interintestinal and intraintestinal variations in human CYP3A dependent metabolism. AB - Cytochrome P450 3A (CYP3A) metabolizes a diverse array of clinically important drugs. For some of these (e.g., cyclosporine, verapamil, midazolam), CYP3A in the intestinal mucosa contributes to their extensive and variable first-pass extraction. To further characterize this phenomenon, we measured CYP3A content and catalytic activity toward the probe substrate midazolam in mucosa isolated from duodenal, jejunal and ileal sections of 20 human donor intestines. For comparison, the same measurements were performed for 20 human donor livers, eight of which were obtained from the same donors as eight of the intestines. Excellent correlations existed between homogenate and microsomal CYP3A content for the three intestinal regions. Median microsomal CYP3A content was greatest in the duodenum and lowest in the ileum (31 vs. 17 pmol/mg of protein). With respect to midazolam 1'-hydroxylation kinetics, the median Km for each intestinal region was similar to the median hepatic Km, approximately 4 microM. In contrast, the median Vmax decreased from liver to duodenum to jejunum to ileum (850 vs. 644 vs. 426 vs. 68 pmol/min/mg). Intrinsic clearance (Vmax/Km) followed a similar trend for the intestinal regions; median duodenal intrinsic clearance was comparable to hepatic intrinsic clearance (157 and 200 microl/min/mg, respectively). Vmax correlated with CYP3A content for all tissues except the ileum. Duodenal and jejunal Vmax and CYP3A content varied by >30-fold among donors. Microsomes prepared from every other 1-foot section of six intestines were also analyzed for CYP3A as well as for two coenzymes. In general, CYP3A activity, CYP3A content and CYP reductase activity rose slightly from duodenum to middle jejunum and then declined to distal jejunum and ileum. Cytochrome b5 content and cytochrome b5 reductase activity varied little throughout the intestinal tract. Regional intrinsic midazolam 1'-hydroxylation clearance was greatest for the jejunum, followed by the duodenum and ileum (144, 50 and 19 ml/min, respectively). Collectively, these results demonstrate that the upper small intestine serves as the major site for intestinal CYP3A-mediated first-pass metabolism and provides a rationale for interindividual differences in oral bioavailability for some CYP3A substrates. PMID- 9400034 TI - Preoperative amiodarone as prophylaxis against atrial fibrillation after heart surgery. AB - BACKGROUND: Atrial fibrillation occurs commonly after open-heart surgery and may delay hospital discharge. The purpose of this study was to assess the use of preoperative amiodarone as prophylaxis against atrial fibrillation after cardiac surgery. METHODS: In this double-blind, randomized study, 124 patients were given either oral amiodarone (64 patients) or placebo (60 patients) for a minimum of seven days before elective cardiac surgery. Therapy consisted of 600 mg of amiodarone per day for seven days, then 200 mg per day until the day of discharge from the hospital. The mean (+/-SD) preoperative total dose of amiodarone was 4.8+/-0.96 g over a period of 13+/-7 days. RESULTS: Postoperative atrial fibrillation occurred in 16 of the 64 patients in the amiodarone group (25 percent) and 32 of the 60 patients in the placebo group (53 percent) (P=0.003). Patients in the amiodarone group were hospitalized for significantly fewer days than were patients in the placebo group (6.5+/-2.6 vs. 7.9+/-4.3 days, P=0.04). Nonfatal postoperative complications occurred in eight amiodarone-treated patients (12 percent) and in six patients receiving placebo (10 percent, P=0.78). Fatal postoperative complications occurred in three patients who received amiodarone (5 percent) and in two who received placebo (3 percent, P= 1.00). Total hospitalization costs were significantly less for the amiodarone group than for the placebo group ($18,375+/-$13,863 vs. $26,491+/-$23,837, P=0.03). CONCLUSIONS: Preoperative oral amiodarone in patients undergoing complex cardiac surgery is well tolerated and significantly reduces the incidence of postoperative atrial fibrillation and the duration and cost of hospitalization. PMID- 9400035 TI - Transvaginal ultrasonography compared with endometrial biopsy for the detection of endometrial disease. Postmenopausal Estrogen/Progestin Interventions Trial. AB - BACKGROUND: Transvaginal ultrasonography is a noninvasive procedure that may be used to detect endometrial disease. However, its usefulness in screening for asymptomatic disease in postmenopausal women before or during treatment with estrogen or estrogen-progesterone replacement is not known. METHODS: We compared the sensitivity and specificity of transvaginal ultrasonography and endometrial biopsy for the detection of endometrial disease in 448 postmenopausal women who received estrogen alone, cyclic or continuous estrogen-progesterone, or placebo for three years. RESULTS: Concurrent ultrasonographic and biopsy results were available for 577 examinations in the 448 women, 99 percent of whom were undergoing routine annual follow-up. Endometrial thickness was less than 5 mm in 45 percent of the examinations, 5 to 10 mm in 41 percent, more than 10 mm in 12 percent, and not measured in 2 percent, and it was higher in the women receiving estrogen alone than in the other groups. Biopsy detected 11 cases of serious disease: 1 case of adenocarcinoma, 2 cases of atypical simple hyperplasia, and 8 cases of complex hyperplasia. Biopsy also detected simple hyperplasia in 20 cases. At a threshold value of 5 mm for endometrial thickness, transvaginal ultrasonography had a positive predictive value of 9 percent for detecting any abnormality, with 90 percent sensitivity, 48 percent specificity, and a negative predictive value of 99 percent. With this threshold, a biopsy would be indicated in more than half the women, only 4 percent of whom had serious disease. CONCLUSIONS: Transvaginal ultrasonography has a poor positive predictive value but a high negative predictive value for detecting serious endometrial disease in asymptomatic postmenopausal women. PMID- 9400036 TI - Cellular adaptations in the diaphragm in chronic obstructive pulmonary disease. AB - BACKGROUND: In patients with severe chronic obstructive pulmonary disease, the diaphragm undergoes physiologic adaptations characterized by an increase in energy expenditure and relative resistance to fatigue. We hypothesized that these physiologic characteristics would be associated with structural adaptations consisting of an increased proportion of less-fatigable slow-twitch muscle fibers and slow isoforms of myofibrillar proteins. METHODS: We obtained biopsy specimens of the diaphragm from 6 patients with severe chronic obstructive pulmonary disease (mean [+/-SE] forced expiratory volume in one second, 33+/-4 percent of the predicted value; residual volume, 259+/-25 percent of the predicted value) and 10 control subjects. The proportions of the various isoforms of myosin heavy chains, myosin light chains, troponin, and tropomyosin were determined by sodium dodecyl sulfate-polyacrylamide-gel electrophoresis. We also used immunocytochemical techniques to determine the proportions of the various types of muscle fibers. RESULTS: The diaphragm-biopsy specimens from the patients had higher percentages of slow myosin heavy chain I (64+/-3 vs. 45+/-2 percent, P<0.001), and lower percentages of fast myosin heavy chains IIa (29+/-3 vs. 39+/ 2 percent, P=0.01) and IIb (8+/-1 vs. 17+/-1 percent, P<0.001) than the diaphragms of the controls. Similar differences were noted when immunohistochemical techniques were used to compare the percentages of these fiber types in the two groups. In addition, the patients had higher percentages of the slow isoforms of myosin light chains, troponins, and tropomyosin, whereas the controls had higher percentages of the fast isoforms of these proteins. CONCLUSIONS: Severe chronic obstructive pulmonary disease increases the slow twitch characteristics of the muscle fibers in the diaphragm, an adaptation that increases resistance to fatigue. PMID- 9400037 TI - Lamotrigine for generalized seizures associated with the Lennox-Gastaut syndrome. Lamictal Lennox-Gastaut Study Group. AB - BACKGROUND: The Lennox-Gastaut syndrome, a severe form of epilepsy that usually begins in early childhood, is difficult to treat. Dose-related drug toxicity is common. METHODS: We conducted a double-blind, placebo-controlled trial of the antiepileptic drug lamotrigine in patients with the Lennox-Gastaut syndrome. Eligible patients had more than one type of predominantly generalized seizure, including tonic-clonic, atonic, tonic, and major myoclonic, and had seizures on average at least every other day. After a 4-week base-line period in which all participants received placebo, we randomly assigned 169 patients (age range, 3 to 25 years) to 16 weeks of lamotrigine (n= 79) or placebo (n=90) in addition to their other antiepileptic drugs. RESULTS: The median frequency of all major seizures changed from base-line levels of 16.4 and 13.5 per week in the lamotrigine and placebo groups, respectively, to 9.9 and 14.2 per week after 16 weeks of treatment (P=0.002). Thirty-three percent of the patients in the lamotrigine group and 16 percent of those in the placebo group had a reduction of at least 50 percent in the frequency of seizures (P= 0.01). There were no significant differences between groups in the incidence of adverse events, except for colds or viral illnesses, which was more common in the lamotrigine group (P=0.05). CONCLUSIONS: Lamotrigine was an effective and well-tolerated treatment for seizures associated with the Lennox-Gastaut syndrome. PMID- 9400038 TI - Images in clinical medicine. Amiodarone-induced skin discoloration. PMID- 9400039 TI - Images in clinical medicine. Amiodarone-induced pulmonary toxicity. PMID- 9400040 TI - House calls to the elderly--a vanishing practice among physicians. AB - BACKGROUND: Despite the growth in other home health care services, the number of house calls by physicians has declined dramatically during this century. We determined the frequency of house calls made by physicians to elderly U.S. patients in 1993 and analyzed the characteristics of the physicians and patients involved. METHODS: We analyzed a 5 percent random sample of the 1993 Medicare Part B claims data for beneficiaries over the age of 65 who were not enrolled in health maintenance organizations (HMOs). With supplemental information from the Area Resource File and the American Medical Association's Physician Masterfile, we determined how many house calls were made, their cost, and a number of specific characteristics of the physicians and the patients. RESULTS: In our 1993 sample, 36,350 house calls were made to 11,917 of the 1,357,262 patients. When extrapolated to all Medicare beneficiaries over age 65 and not enrolled in HMOs, these figures correspond to 727,000 house calls to 238,340 patients nationwide. We estimated the cost of these house calls to be $63 million. The patients who received house calls from physicians were older than those who did not, were more likely to die within the calendar year, had higher rates of hospitalization, and were more likely to receive care from other home health providers, hospice programs, and skilled-nursing facilities. Patients residing in rural areas and those in areas with high physician-to-population ratios had an increased likelihood of receiving a house call. The physicians who made house calls were more likely than others to be generalists, osteopaths, older, male, board certified, practicing in the Northeast, and in solo practice. CONCLUSIONS: A very small percentage (0.88 percent) of elderly Medicare patients, mainly those who are very sick and near the end of life, receive house calls from physicians. PMID- 9400041 TI - Transmissible spongiform encephalopathies. PMID- 9400043 TI - Optimizing the accuracy of transvaginal ultrasonography of the endometrium. PMID- 9400044 TI - Can house calls survive? PMID- 9400045 TI - Inappropriate drug-donation practices in Bosnia and Herzegovina, 1992 to 1996. PMID- 9400046 TI - An unusual congenital anomaly: ectopic sigmoid kidney combined with hermaphroditism in a newly born calf. AB - A rare case of hermaphroditism accompanied with ectopic sigmoid kidney in cross bred calf is reported. Findings revealed fused kidneys located near urinary bladder, and presence of uterus, vagina, penis and testicles. Both urinary and genital defects seemed to occur in combination and to be interrelated. PMID- 9400047 TI - Observations on the fine structure of the liver in the camel (Camelus dromedarius). AB - The structure of macroscopically inconspicuous livers in 23 adult camels (Camelus dromedarius) was studied by light and transmission electron microscopy. A well developed connective tissue characterizes the camel liver. Thick trabeculae divide the liver parenchyma into lobules. Portal tracts and central veins are surrounded by a variable amount of fibrous tissue. In the perisinusoidal space (DISSE), collagen fibres form a dense three-dimensional network around the sinusoids. A mild to moderate fatty infiltration is present in hepatocytes of all animals. In the epithelial cells of the bile ducts, small to medium sized lipid inclusions are a common feature. The ultrastructure of hepatocytes in the camel liver corresponds to that of other domestic mammalian species. The endothelial cells lining the sinusoids show a multiple fenestration and are surrounded by a discontinuous basal lamina. Fat-storing cells are numerous and contain lipid droplets varying in size, number and electron density from one cell to another. PMID- 9400048 TI - Elastic fibre system of the female canine urethra. Histochemical identification of elastic, elaunin and oxytalan fibres. AB - The elastic system fibres of the female urethra were investigated in seven dogs of different breeds. The fibres were stained and differentiated with orcein, Verhoeff's iron hematoxylin, and pararosanilin-based aldehyde fuchsin, with and without previous oxidation. Orcein and aldehyde fuchsin revealed some subepithelial longitudinally orientated delicate fibres (elaunin fibres) and numerous coarse longitudinal fibres (elastic fibres) in the deeper subepithelial connective tissue containing the vascular plexus, as well as a network of fibres of different calibers in the periurethral connective tissue. Elastic system fibres were also found in association with the sinusoids of the vascular plexus and the urethral smooth musculature. Verhoeff's iron hematoxylin only reacted distinctly with coarse (elastic) fibres. When applied following oxidation, aldehyde fuchsin disclosed an extensive meshwork of additional delicate fibres (oxytalan fibres) in the subepithelial connective tissue adjacent to the basement membrane. Oxytalan fibres were also discernible in the sinusoidal adventitia of the vascular plexus, as well as between smooth and striated muscle fibres. Due to the predominantly longitudinal orientation of the elastic system fibres and the low urethral resistance to manual expression of urine from the bladder post mortally, when the elasticity of the urethra is still intact and the activity of other continence factors can be excluded, the elastic tissue is not judged to be a major contributory factor to urinary continence. PMID- 9400049 TI - Histochemical demonstration of lipase activity in the gastric mucosa of the cat. AB - The aim of the present study was to determine, histochemically, the onset and location of production of preduodenal lipase in fetal, suckling, weaned and adult cats. Strong enzymatic activity was localized in the surface mucous cells of the gastric mucosa in animals at postpartal day 1 after ingestion of milk. Activity of gastric lipase persisted as long as animals were nursed. No gastric lipase could be demonstrated in weaned and adult cats. Lingual lipase was not found at any developmental stage examined. Thus, in the newborn cat, lipase of the gastric mucosa is responsible for milk fat lipolysis. PMID- 9400050 TI - A stereological study of the different cell populations in chicken testes treated with follicle-stimulating hormone during embryonic development. AB - Quantitative morphological methods were used to analyse the histomorphometric changes and variations in the number and size of cells from diverse cellular populations in testes of newly hatched chicks treated with follicle stimulating hormone (FSH) during embryonic development. The tissue was fixed and embedded in Epon and sections were morphometrically measured under light microscopy, using point counting for volume densities and the Floderus equation to determine numerical density. The average volume of the individual cell was determined by dividing the volume density by the numerical density. Results indicate that FSH administration causes an increase in the number of Sertoli cells and spermatogonia, as well as enlargement of the individual Sertoli cells leading consequently to an increase in the diameter and volume density of the testicular seminiferous tubules. Results also reveal an increase in the volume density of the interstitial cords of the Leydig cells, this expansion is due to the enlargement of the individual Leydig cells and not to an increase in their number, which remains constant. We conclude that testes of chick embryos are able to respond to FSH treatment, as revealed by the changes in the number and size of the cells conforming the diverse cellular populations of the testis. FSH treatment during embryonic development induces histomorphometric changes in both the interstitial tissue and seminiferous tubules, accelerating their growth and differentiation. PMID- 9400051 TI - Ultrastructural study on the stomach of Tilapia spp (Teleostei). AB - An ultrastructural study has been made of gastric mucosa of a teleostean fish, Tilapia spp. The cytological features of the surface mucous cells, mucous neck cells, glandular cells and endocrine cells are described. The surface mucous cells, identified by their superficial localization, are characterized by apical granules. The mucous neck cells are distinguished by the appearance of their mucous granules and their localization between surface mucous cells and glandular cells. The gastric glands contain only one form of cell whose fine structure is similar to cells that secrete hydrochloric acid. Physiological implications of some ultrastructural features are also discussed. PMID- 9400052 TI - The annual testicular cycle of the domestic quail (Coturnix coturnix japonica). AB - A morphometric study was conducted on the testis of the domestic quail Coturnix coturnix japonica to determine testicular kinetics. We investigated the variability along the year of testicular parameters such as seminiferous tubule diameter, germinal epithelium height and amount of meiotic figures of maturing spermatids in the seminiferous epithelium and of sperm in the tubular lumen. The results of morphometric analysis showed the occurrence of an annual testicular cycle defined by four distinct phases: a resting phase (at the end of summer), a recrudescence phase (in the fall), a proliferative phase (at the end of winter and beginning of spring), and a regression phase (spring and summer). We also observed that the testes of adult quails present elevated and maximal spermatogenic activity in fall-winter (short-day period) and at the beginning of spring, respectively, and lower values in spring and summer (long-day periods), with minimum values at the end of summer. PMID- 9400054 TI - Torsion of accessory spleen in an adult patient: imaging findings at CT, MRI and angiography. AB - A 40-year-old woman presented with acute left upper quadrant pain due to torsion of an accessory spleen around its long vascular pedicle, causing infarction. Torsion of an accessory spleen is extremely rare. As far as we known only 14 cases have previously been reported in the literature, and more than half patients were children. MRI can be helpful in the differential diagnosis of infarction by suggesting haemorrhagic necrosis on the T2-weighted images. PMID- 9400053 TI - Safety and efficacy of contrast-enhanced MRI in the brain, head and neck: gadodiamide injection versus gadopentate dimeglumine. AB - The objective of the present study was to evaluate the safety and efficacy of gadodiamide injection, a non ionic MRI contrast medium in comparison with the ionic agent gadopentate dimeglumine. Two groups of 50 patients with known or suspected lesions of the brain or head and neck were enrolled in a double -blind, randomised trial. In the gadopentate dimeglumine group three patients reported four adverse events, and in the gadodiamide injection group, four patients reported four side effects. All events were minor. Two radiologists analyzed pre and post-contrast MR images. The parameters evaluated were the number of lesions, delineation of the lesion, gain of diagnostic information, and final diagnosis. Both contrast media gave identical diagnostic information. PMID- 9400055 TI - Shock bowel following massive pulmonary embolism. AB - Wide-spread abnormalities of the small bowel on CT scan after massive pulmonary embolism and acute hemodynamic collapse are described. These small bowel abnormalities are secondary to hypotension with prolonged hypoperfusion. They consist of diffuse thickening of the small-bowel wall, fluid-filled, dilated loops and increased contrast enhancement of the small-bowel wall (shock bowel). These abnormalities are reversible and should be distinguished from acute vascular occlusion. PMID- 9400056 TI - Pseudomyxoma peritonei in association with primary malignant tumor of the ovary and colon. AB - Pseudomyxoma peritonei (PMP) is a rare entity that is characterised by abundant intraperitoneal mucinous and gelatinous material associated with an intraperitoneal adenocarcinoma. We report the case of a patient who presented with PMP associated with a ruptured well-differentiated mucinous adenocarcinoma of the ovary and an infiltrating moderately-differentiated mucinous adenocarcinoma of the sigmoid. Diagnosis was made by ultrasonography and CT. Due to the presence of 2 mucinous tumors with different histological grade the most likely pathogenesis was that of multifocal metaplasia. The ovarian and colonic mucinous tumors were independent primary neoplasms and PMP probably was the result of rupture of one of these tumors with peritoneal seeding of viable mucus secreting tumor cells. Aggressive surgical debulking in addition to left hemicolectomy and radical hysterectomy were performed. PMID- 9400057 TI - Pericystic emphysema in pulmonary hydatid disease. AB - The case of a 12-year-old Caucasian boy, living in a non-endemic area, with two intrapulmonary masses is presented. Conventional X-rays and computed tomography images were highly suggestive of pulmonary Echinococcus disease because of the presence of pericystic emphysema in one of both masses, a finding known as the "crescent" or "meniscus" sign. This radiological feature and other highly suggestive imaging findings of pulmonary hydatid disease are presented and discussed. PMID- 9400058 TI - Giant cell tumor of the tendon sheath in the knee. AB - The clinical, imaging and pathologic findings in a 28-year-old patient with a giant cell tumor of the tendon sheath in the knee are reported. PMID- 9400060 TI - Benefits of a new direct digital x-ray imaging system. PMID- 9400059 TI - MR imaging of bone infarction and epiphyseal osteonecrosis. AB - No single musculoskeletal disorder has generated more passionate discussion than bone osteonecrosis. Surprisingly, its physiopathogenesis remains largely unknown despite in-depth studies. Its treatment is still debated, and its wide range of clinical manifestations which vary from totally asymptomatic to the catastrophic event of irreversible epiphyseal collapse remains a fascinating question for the clinicians (1-4). Magnetic Resonance (MR) imaging has given a large tribute to the discussion during the last decade by allowing detection of marrow infarcts at a presymptomatic period of the disease, thus providing data on its natural history. Unfortunately, MR imaging has also contributed to increase the confusion among various epiphyseal disorders. The aim of the current paper is to provide an overview of the current knowledge of imaging features by stressing accepted data and delineating blind areas. PMID- 9400061 TI - Three-dimensional strain analysis of the human left ventricle. PMID- 9400062 TI - Fast volumetric data acquisition in abdominal computed tomography and magnetic resonance imaging. PMID- 9400063 TI - Some preliminary studies on the ability of Africanized honey bees (Apis mellifera L.) to tolerate cold temperatures when placed inside a refrigerator. AB - Cold is often suggested as an ecological mechanism to prevent the migration of Africanized honey bees. The ability of Africanized honey bees to tolerate cold temperatures was investigated. In one study an observation hive was placed inside a refrigerator at 25 degrees C. The study was conceptualized as a choice experiment in which the colony could remain in a cold environment or leave for a warm environment. Analysis indicated that the bees remained at 9 +/- 1 degrees C for 14 days before leaving. In a second series of studies, testing the tolerance to 0 degree C, 280 bees were placed individually in small metal tubes. The data gathered included survival rate, time to regain consciousness, and ability to feed. Analysis indicated that Africanized bees can survive for up to 3 hr. at 0 degree C with few ill effects. At 4 hr., however, the survival rate is low. Limitations of the study, the use of cold as a possible deterrent to honey bee mites, and suggestions for additional research are discussed. PMID- 9400064 TI - Racial attitudes of preschoolers: age, race of examiner, and child-care setting. AB - Racial attitudes of 60 preschool children (28 boys, 32 girls) from either a monoracial Euro-American child-care program (n = 16), a monoracial African American program (n = 12), or a multiracial program (25 Euro-Americans, 7 African Americans) were assessed using the Preschool Racial Attitudes Measure II. Despite the over-all neutral attitudes reflected by these children, evidence of a Euro American bias among older children was found. If replicated with a large randomly selected sample recognizing and understanding early racial attitudes may be a key factor in fostering positive racial identity and preventing the formation of prejudice. PMID- 9400065 TI - A prospective study of hope, optimism, and health. AB - The present investigation sought to distinguish hope from optimism in the context of a 10-wk. prospective study involving reports of health outcomes. Gottschalk's (1985) Hope Scale and Scheier and Carver's (1987) Life Orientation Test which assesses optimism were given to subjects, along with a health questionnaire. Ten weeks later subjects were given a second health questionnaire. To rule out potential confounds we included measures of neuroticism, depression, extroversion, and social desirability. After controlling for the effects of correlated confounds, we found that lower hope scores (but not optimism) were correlated with several dimensions of reported health, including frequency and severity of illness. PMID- 9400066 TI - Patients' task-orientation and perceived benefit of amplification in hearing impaired elderly persons. AB - This study investigated the reported handicap of 50 elderly hearing-impaired patients who were classified as high or low in task-orientation. Measures of perceived handicap were taken when subjects were fitted with hearing aids and three months later. Analysis indicated that subjects classified as both high and low on task-orientation reported significant increases in their hearing impairment at 3 mo., but subjects classified as high on task-orientation reported significantly less handicap than subjects classified as low. PMID- 9400067 TI - Note on an MMPI-2 scale of early sexual abuse. AB - The mean of 191 female psychiatric inpatients on the Griffith, Myers, and Tankersley (1996) MMPI-2 scale of childhood sexual abuse was compared with means of 2 community samples (58 sexually abused women and 57 nonabused women). The mean of the patients was substantially larger than that of the nonabused women but slightly larger than that of the abused women. The scale may measure general maladjustment or psychopathology instead of, or in addition to, specific sequelae of sexual abuse. Further investigation is necessary to cross-validate the scale in community samples and to examine whether scores differentiate abused and non abused women in clinical samples. This note illustrates use of an archival data set with results of recent research. PMID- 9400068 TI - Dyskinesias subside off all medication in a boy with autistic disorder and severe mental retardation. AB - A boy with autistic disorder and severe mental retardation developed severe dyskinesias, including objective akathisia (probable) and tics, a month after discontinuation of at least two years of treatment with drugs block dopamine receptors. These dyskinesias greatly subsided during a 17-wk. open-label nonblind clinical trial of clomipramine, and returned transiently when the parents abruptly discontinued clomipramine. However, the dyskinesias gradually subsided during two and a half years of follow-up with the boy being off all medication. A few stereotypies remain. We believe this suggests the hypothesis that movement disorders, such as withdrawal and tardive akathisia and tics, occurring in boys with autistic disorder treated with dopamine receptor-blocking drugs may subside months or years after discontinuation of the agents and that clomipramine may facilitate this process. We also hypothesize that some boys with autistic disorder and mental retardation exhibit fewer movement disorders, fewer psychiatric symptoms, and better over-all functioning after they have received no dopamine receptor-blocking drugs for several months, and this improvement continues years after the medication has ceased. PMID- 9400069 TI - Achromatic visual backward masking of colored stimuli in type I diabetes. AB - On a visual backward masking task using color stimuli with an achromatic patterned mask, we compared the masking performances of 3 Type I diabetics with those of 9 participants in a control group. Analysis indicated that the diabetics show a marked decrement in performance with blue stimuli and a lesser decrement with red stimuli. Suggestions for further theoretical and parametric studies are discussed. PMID- 9400070 TI - Locus of control of smokers, nonsmokers, and nonpracticing smokers. AB - In this survey, score analyses of 123 male and female respondents, ages 21 to 33 years, yielded no significant differences between either sex and smokers versus nonsmokers on Rotter's locus of control scale. Of particular interest was that nonpracticing smokers (quitters) scored more internal than either smokers or nonsmokers. PMID- 9400071 TI - Fear of AIDS and Homophobia Scales: additional estimates of reliability and validity. AB - Psychometric properties of the Fear of AIDS Scale and the Homophobia Scale from 85 heterosexual undergraduates (and 44 gay, lesbian, and bisexual undergraduates) were estimated. Responses on the Homophobia Scale were internally consistent (alpha = .88), had 4% standard error of measurement, and scores differed by subjects' sexual orientation. Responses to Fear of AIDS Scale were moderately internally consistent (alpha = .75), had 12% standard error, and were significantly different for the two groups. Scores on the tests were correlated .66. Researchers must assess the relationship between scores on these measures. PMID- 9400072 TI - Comparison of scores on the MMPI-A and MMPI-2 for young adults. AB - Male college students' profiles look more pathological on the adult version of the MMPI than on the adolescent version. In the present study, men showed elevated scores on the F, Pa, and Sc scales on the MMPI-2. In contrast, women's profiles were more normal on the adult version. When designing the MMPI-A, the authors attempted to maintain correspondence with the original MMPI and the MMPI 2 so the scales could be interpreted similarly. This study compared scores on the MMPI-A and MMPI-2 by administering both tests to the same subjects (N = 43; 19 men and 24 women). Validity and standard scale scores were compared using Pearson product-moment correlations (r), T-score means and standard deviations, and average profiles. Codetype comparisons were also made. In general, MMPI-A and MMPI-2 codetype analyses did not agree. The codetype approach is not recommended for interpretation of the MMPI-A. The finding that young college men show elevated scores on MMPI-2 scales is consistent with previous research and suggests that the MMPI-A may be a useful tool for 18-yr.-old men. PMID- 9400073 TI - Loneliness and sexual risk behavior in gay men. AB - This study examined loneliness in a sample of gay men and its association with unprotected anal intercourse, social support, instability of self-esteem, intimacy, and coping. A sample of 470 urban gay men completed a self-administered questionnaire. Participants scored high on Loneliness in comparison to other samples. Measures of Intimacy, Social Support, Instability of Self-esteem, monogamous relationship status, and use of Avoidance Coping predicted 58.5% of the variance in Loneliness scores. Both social and psychological variables appear to be important for understanding loneliness in this population. Men who had unprotected anal intercourse with nonprimary partners during the previous six months scored higher on Loneliness than other participants, but those who did so with primary partners scored the lowest. Episodes of unprotected anal intercourse with nonprimary partners might have been Avoidance strategies to help participants cope with loneliness or or other negative affect. PMID- 9400074 TI - Personality correlates of religiosity among adults in the Republic of Ireland. AB - This study examined the relationship between measures of personal religiosity (religious attitude, frequency of personal prayer), a measure of public religiosity (church attendance), and the Abbreviated form of the Revised Eysenck Personality Questionnaire among 216 adults in the Republic of Ireland. A significant negative correlation was found between scores on psychoticism and on the three measures of religiosity among men (religious attitude, -.36; frequency of personal prayer, -.40; and frequency of church attendance, -.30) and among women (religious attitude, -.40; frequency of personal prayer, -.47; and frequency of church attendance, -.31). No significant relationship was found between any of the religiosity measures and the other measures contained within the Eysenck scores. A further analysis of the data suggests that the relationship for measures of public religiosity with low psychoticism is only a facet of the relationship between public and personal religiosity. These findings add to a growing body of research which locates religiosity within the psychoticism dimension of Eysenck's model of personality and adds to prior suggestions that this approach is applicable only to personal aspects of religiosity. PMID- 9400075 TI - Duty-related stressors and PTSD symptoms in suburban police officers. AB - This study examined the effects of duty-related stress on police officers. Using a sample of 100 suburban police officers, an anonymous questionnaire requested demographic information and included a measure of duty-related stressors, SCL-90 R, the Posttraumatic Stress Disorder scale of the Impact of Events Scale-Revised, and a locus of control scale. Also assessed was whether Critical Incident Stress Debriefing was experienced. The results showed significant correlations between scores on duty-related stress, somatization, and symptoms of PTSD. 13% of the sample met the DSM-IV (1994) diagnostic criteria for PTSD. Results of the regression analysis showed the best predictors for the diagnosis of PTSD were associated with the factor of Exposure to Death and Life Threat, which corresponds to the DSM-IV AI criteria. Finally, 63% of the respondents stated that a critical incident debriefing would be beneficial following an extremely stressful event related to duty. PMID- 9400076 TI - Is sex of fetus associated with duration of labor in nulliparas? AB - In a post facto study to examine whether sex of fetus was related to duration of nulliparas' labor 30 nulliparous (had not given birth previously) women who delivered boys and 30 others who delivered girls were matched according to the criteria that they were nulliparous, received an epidural for analgesia during labor, and their labors were either induced or augmented. All delivered vaginally. Duration of labor was not statistically significantly related to sex of the fetus. PMID- 9400077 TI - Definition and measurement of affective variables: theoretical and methodological considerations. AB - Research in the affective domain is handicapped by the absence of a theory to guide empirical work. Without a theoretical framework to guide the selection of variables, to design appropriate measures, and to interpret results, research consists of a set of discrete studies that have no collective impact. PMID- 9400078 TI - Parental divorce and narcissism among college students. AB - 342 women and 225 men, undergraduate students, participated in a study to assess whether experiencing the divorce of one's parents affected narcissistic development. In a larger study on the long-term effects of divorce, these students completed the Narcissistic Personality Inventory. The analyses indicated that the scores for children from divorced families did not differ from the scores of children from intact families on any of the seven subscales. PMID- 9400079 TI - Testing the McBeath hypothesis: relation of sexual orientation and belief in the paranormal. AB - According to McBeath, the incidence of homosexuality among experiments of paranormal phenomena exceeds that which would be expected by chance. Therefore, 50 homosexual men and 50 heterosexual men were administered the forced-choice version of the 18-item Australian Sheep-Goat Scale as a measure of belief in and alleged experience of the paranormal. As no differences were found in scores on belief/experience, there was no evidence for McBeath's hypothesis. PMID- 9400080 TI - Effects of reading motivation on the belief in and consumption of newspapers among youth. AB - A motivational model of newspaper consumption was elaborated. In this model, reading motives are supposed to generate certain beliefs in newspapers. The belief in the satisfying properties of a newspaper is based on an evaluation of how well the attributes of a newspaper are expected to match the motives of individuals. Once a match between the motives and the belief is established, then motivation is triggered, and newspaper consumption should occur. We tested the model with the hypotheses that reading motives explain belief in newspapers to a greater extent than the consumption of them and that beliefs explain consumption to a greater extent than reading motives. Reading motives, beliefs and consumption of general broadsheet, business and tabloid newspapers were measured in 1343 young people between the ages of 15 and 25. The results supported the hypotheses. The results indicate that the profiling and development of the newspapers toward more loyal and new readers should be based on the readers' beliefs in newspapers and the motives explaining them. PMID- 9400081 TI - Anxiety and confidence in using a library by college freshmen and seniors. AB - The first study using a measure of library anxiety showed that 180 college freshmen have significantly higher scores than college seniors. A second study found that among freshmen students, those with high scores on Library Anxiety reported less confidence in using the library than those with low scores, even though the students were equal on several indices of academic ability and performance. PMID- 9400082 TI - Environmental factors and clients' self-disclosure in counseling. AB - Self-disclosure by clients is regarded as a fundamental component of counseling, with increased self-disclosure being related to positive outcome of treatment; however, scant attention has focused on environmental characteristics that may facilitate self-disclosure among clients. An evaluation of prior research in this area leads to suggestions for studies on environmental factors and self disclosure. PMID- 9400083 TI - Psychology of computer use: XLVIII: Relation between playfulness and computer anxiety. AB - Correlations between scores for computer anxiety and for playfulness were investigated (N = 265). Scores on computer anxiety correlated negatively with overall scores on the playfulness scale and the factors, Fun and Creative. Only the correlation with scores on Creative remained significant when control for computer experience was imposed. The results imply that playfulness relates to computer anxiety indirectly through its relation with computer experience. The implications of the results for the validity of the Adult Playfulness Scale are briefly discussed. PMID- 9400084 TI - A validity study of the MMPI-TRI Acting-out Scale. AB - 49 college men and 45 women were administered the 1995 MMPI-TRI by Swanson, Templer, Thomas-Dobson, Cannon, Streiner, Reynolds, and Miller and a short form of the MMPI with scales for Subjective Distress, Acting-out, and Psychosis. To test the validity of the Acting-out scale respondents also took measures of sexual and aggressive acting out as well as a measure of alcohol use. They were asked about their use of drugs. Women had significant correlations between scores on the Acting-out Scale and scores on measures of Sexual Sensation Seeking .57, Sexual Compulsion .54, Anger Control -.40, Anger-out .50, Anger-in .32, and Drug Use .40. Men had significant correlations for scores on measures of Sexual Compulsion .51, Anger Control -.39, Anger-in .37, and Alcohol Use .35 but not Anger-out. Results suggest the Acting-out scale is more valid for college women than for men. PMID- 9400085 TI - An informational versus monetary incentive in increasing physicians' response rates. AB - This study examined return rates for a cancer prevention survey by pediatricians in relation to an informational booklet versus a monetary incentive in the first of a three-wave mailing. Of the 300 surveys sent which included an informational booklet incentive, 189 (64%) were returned. Of the 300 surveys sent which included a $1.00 incentive 227 (79%) were returned, indicating the $1.00 incentive was more effective than the informational incentive in increasing return rates in this sample of physicians. PMID- 9400086 TI - Religiosity and psychological disturbance. PMID- 9400088 TI - Social anxiety and performance in an interpersonal perception task. AB - The Interpersonal Perception Task-15 videotape served as a criterion measure to test hypotheses about individual differences in interpersonal perception. 160 undergraduates completed the Personal Report of Communication Apprehension Scale, the Shyness and Sociability Scale, and the Interpersonal Perception Task-15. Scores on the Communication Apprehension Scale were negatively correlated with Interpersonal Perception Task-15 scores, as predicted. Scores on the Shyness scale were negatively correlated with scores on the Interpersonal Perception Task 15, while Sociability scale scores were positively correlated. These results underscore the association between social anxiety and interpersonal perception. PMID- 9400087 TI - An MMPI-2 scale to identify history of sexual abuse. AB - This study attempted to extract an MMPI-2 scale identifying adult male and female victims of severely traumatogenic childhood sexual abuse. Such a scale might aid in the early detection and diagnosis of trauma sustained from abuse and lead to prevention of childhood sexual abuse in subsequent generations since victims are more likely to expose their offspring to similar abuse. Victims of severe childhood sexual abuse almost inevitably suffer from trauma. Taking the severity of abuse into consideration, a higher identification rate than the one obtained in previous attempts was expected. The sample (N = 201), recruited from a low-fee outpatient clinic and a community mental health center, was divided into a severe childhood sexual abuse-reporting group and a control group, based on Russell's (1983) definitions. To cross validate, the data were analyzed three ways. A stepwise discriminant function analysis based on Wilks Lambda yielded 95% correct classification (using 52 MMPI-2 items), a discriminant function analysis based on chi-squared yielded 77% classification (using 63 items), and a stepwise logit analysis yielded 71% classification (using 15 items). There was little agreement among the items identified by the three solutions. PMID- 9400089 TI - Threats and anti-semitic blaming. AB - A strategy for efficiently and effectively reducing anti-Semitic stereo-typing was presented, supported by the results of a questionnaire responded to by 15 randomly selected students between the ages of 18 and 32 years, at L.I.F.E. Bible College (associated with the Pentacostal Four-Square Church) in San Dimas, CA in 1993. Relationships predicted among stereotypic blaming and related threats were supported by the data--principally, that very negative combinations of common blamings called compound blamings, e.g., "Jews are rich because Jews are more dishonest," correlated significantly with a large group of far less negative anti Semitic blamings and related threats. It was argued that a single compound blaming when deconditioned in a classroom, would be more effective in reducing over-all anti-Semitic blaming than deconditioning any other blaming or by using the more traditional group discussion or lecture methods for reducing prejudicial attitudes. PMID- 9400090 TI - APA, science, and the Defense of Marriage Act. AB - This paper uses the APA position on the Defense of Marriage Act to demonstrate that APA's advocacy policy regarding homosexual marriage is not based on science. It shows that the politics of advocacy have led a purportedly scientific organization to misinterpret, overgeneralize, and distort the results of research and to ignore the original purpose of the organization. PMID- 9400091 TI - Inducing positive mood without demand characteristics. AB - The possibility that a state of private self-awareness induced by a small mirror can facilitate the effect of procedures which induce a positive mood was investigated. Participants were 171 female and 60 male undergraduates who were randomly assigned to one of six conditions in a 2 (Mirror vs No-mirror) x 3 (Control vs Velten manipulation vs Music manipulation) design. As predicted, participants who experienced the Velten and Music manipulations in the presence of the mirror reported elevated mood relative to control participants. The mood of participants who experienced the Velten and Music manipulations without the mirror did not differ from the mood of control participants. The potential benefits of using a small mirror as a substitute for explicit instructions about the expected effect of mood-induction procedures are discussed. PMID- 9400093 TI - The prevalence of lung lesions in pigs at slaughter in Switzerland. AB - A survey of the prevalence of lung lesions in randomly selected slaughtered swine representative of the Swiss fattening pig population was carried out from May to September 1992 in six large abattoirs. In total 8921 lungs out of 561 herds were examined. Histological investigations were completed in every herd. No gross lesions could be found in 56% of the pigs. The most frequent lesions in individuals were bronchopneumonia (21%) and diffuse pleuritis (21%). Linear scars (9%), focal pleural fibrosis without any pneumonic lesion (2%), pleuropneumonia (1%) and abscesses (1%) were less prevalent. The prevalence of lesions at the herd level was completely different. The lungs of only 14% of the herds were free of any lesions. The main finding at the herd level was a diffuse pleuritis (75%), followed by bronchopneumonia (64%), linear scars (60%), focal pleural fibrosis (20%), abscesses (14%) and pleuropneumonia (9%). In 94% of the herd samples with macroscopically diagnosed bronchopneumonia, the histological lesions were consistent with enzootic pneumonia of pigs. PMID- 9400092 TI - The effect of fenarimol on marker enzymes in rat liver in two-stages model of hepatocarcinogenesis. AB - The two-stage model for the development of early markers of hepatocarcinogenesis was applied to assess the potential of fungicide fenarimol (alpha-(2 chlorophenyl)-alpha-(4-chlorophenyl)-5-pirimidinemethanol++ +) as a possible promoter in this process. In this experiment the rats were subjected to partial hepatectomy (to induce proliferation), followed by the single (50 mg/kg bw) dose of diethylnitrosamine (DEN) (initiator) and then, followed by the 26 weeks exposure to fenarimol administered in the olive oil suspension (250 mg/kg daily). The activities of gamma-glutamyltransferase (GGTase), glucose-6-phosphatase (G-6 Pase) and alkaline phosphatase (APase) regarded as markers of early stages of hepatocarcinogenesis were measured biochemically and histochemically in the livers of exposed rats as well as in the respective positive and negative controls. Rats exposed to 2-acetylaminofluorene (2-AAF), instead of fenarimol, served as positive controls. It was found that in the full initiation/promotion regimen both 2-AAF and fenarimol induced the increase of GGTase activity in the liver and formation of GGTase-positive hepatocytes. However the exposure to fenarimol alone also increased GGTase activity, although this response was not observed in rats exposed to 2-AAF alone. The possible mechanisms and explanation for such types of responses were discussed, and conclusion has been drawn that fenarimol did not affect the rat hepatocarcinogenesis induced by PH and DEN. PMID- 9400094 TI - ANA hails passage of Medicare reimbursement. PMID- 9400095 TI - Health and safety on the job: nurse, protect thyself. PMID- 9400097 TI - Colorado case blurs line between board of nursing admin. law and criminal action. PMID- 9400096 TI - Long-awaited Providence ruling upholds right of charge nurses to bargain. PMID- 9400098 TI - Nursing ethical code reflects changing times. PMID- 9400099 TI - SNAS key in securing Medicare reimbursement for NPs, CNSs. PMID- 9400100 TI - Arkansas, Louisiana partner to address membership issues. PMID- 9400101 TI - ONA intensifies campaign against hazardous powdered latex gloves. PMID- 9400102 TI - ANA quality indicators applied to unit research. PMID- 9400103 TI - Nurses lead response to flooding in North Dakota and Minnesota. PMID- 9400104 TI - Western approaches--Part 2. PMID- 9400105 TI - The child in theatre: should parents be involved? AB - In this article the author will discuss the parental role in relation to children undergoing surgery. The gap between theory and practice in relation to this issue is recognised, and an inter-relationship which must occur has been described (McCaugherty 1991). The principles of theory are valuable, but it is only through application to practice that problems can be properly identified. Equally, the ability to deal with such problems as they arise can only occur when the underpinning knowledge of theory enables foresight. The author will examine theories and research into parental involvement in their child's care whilst the child is in the operating department and discuss problems which have arisen during application to practice. PMID- 9400106 TI - The perioperative nurse--carer or technician? PMID- 9400107 TI - NATN/3M Award Winner. Benchmarks. PMID- 9400108 TI - Universal precautions and infection control in the perioperative setting. PMID- 9400109 TI - Homeostasis--the key concept to physiological control. AB - The maintenance of body water content is a classic example of homeostasis at work. Water is continuously lost and added to the systems. The regulation of a balance between the factors involved demonstrates the dynamic nature of homeostatic processes. Surgery places additional demands on such processes, partly because there are additional factors in the balance equation and partly because of the hormonal responses to trauma which also affect water balance. Promoting the return to a balance state and maintaining it, during and after surgery, is important to patient well-being and may even facilitate recovery. The risks associated with a disturbance in water balance are of potentially greater consequence if there is water overload, particularly if the patient has underlying cardiovascular or respiratory problems. Slight dehydration is probably a better target to aim for in order to reduce such risk but there are no easy ways to achieve this state as individuals will vary in their responses to surgery. The hydration, and electrolyte, needs, will vary between patients so fluid therapies should be individualised. Whilst a patient's fluid balance chart provides a means of assessing water balance, the interpretation is complicated after surgery. An awareness of other signs is therefore essential. PMID- 9400110 TI - Alison Bell Memorial Writers Award. Creutzfeldt-Jakob disease--an emerging risk. PMID- 9400111 TI - The shortage of theatre personnel--a crisis! PMID- 9400112 TI - Pet therapy ... a howling success. PMID- 9400113 TI - New guidelines for cardiac rehab. PMID- 9400114 TI - An update on prescriptive authority. PMID- 9400115 TI - Professional boundaries in nursing. PMID- 9400116 TI - Consumer Knowledge: the key to managed care success. PMID- 9400117 TI - Nursing education is key to reducing organ and tissue shortage. PMID- 9400119 TI - Nursing certification ... a student's perspective. PMID- 9400118 TI - A job no one else can do. PMID- 9400120 TI - The memories of a mentor. PMID- 9400121 TI - Health needs impact academics. PMID- 9400122 TI - The nurse as witness: depositions. PMID- 9400123 TI - Workplace issues. PMID- 9400124 TI - Healthcare outcome studies: eliminating the "yes but" factor. PMID- 9400125 TI - Nurses caring for nurses. PMID- 9400126 TI - You can't have a better employee than a recovering nurse. PMID- 9400127 TI - Supervision of and delegation to UAPs. PMID- 9400128 TI - Perceptions of parent and nurse relationships and attitudes of parental participation in caring for infants in the NICU. PMID- 9400130 TI - Nursing pins: passe or proof. PMID- 9400129 TI - Report of survey results: the 1995 Kentucky Nurses Association Workplace Survey. PMID- 9400131 TI - Moving forward: advanced practice psychiatric/mental health nursing. PMID- 9400132 TI - Show me the way to go home. PMID- 9400133 TI - Kentucky Board of Nursing. Advisory opinion statement. Roles of nurses in the supervision and delegation of nursing acts to unlicensed personnel. PMID- 9400134 TI - Are you a negligent delegator? PMID- 9400136 TI - KNA takes patient safety to Frankfort. PMID- 9400135 TI - The Kentucky Board of Nursing and the Kentucky Nurses Association: what's the difference? A comparison of the organizations. PMID- 9400137 TI - What's it like to have an angioplasty? PMID- 9400138 TI - Nurses need collective bargaining collective bargaining is professional. PMID- 9400139 TI - Follow-up of intracoronary stent patients. PMID- 9400140 TI - Perkins Center offers comprehensive vocational rehabilitation. PMID- 9400141 TI - Workplace woes. PMID- 9400142 TI - Collaboration and nurse anesthesia. PMID- 9400143 TI - Defining collaboration. PMID- 9400145 TI - [Starting in Cologne: the initiative: chronic wounds. Guideline: decubitus ulcer]. PMID- 9400144 TI - Policymaking and politics: through the eyes of nursing students. PMID- 9400146 TI - [Thr initiative "chronic wounds"--reasons and aims]. PMID- 9400147 TI - [The economic situation of chronic wounds]. PMID- 9400148 TI - [Renaissance of infectious diseases. Which infections are of importance today?]. PMID- 9400150 TI - [Health care expenses--an international comparison]. PMID- 9400151 TI - [Early diagnosis of onychomycosis. The less severe, the better are the chances for a cure]. PMID- 9400149 TI - [Patients with vascular diseases--their care in theory and practice. Living with the disease--everyday experiences at the department]. PMID- 9400152 TI - [Training of auxiliary personnel--an important contribution to the relief of registered nurses]. PMID- 9400153 TI - [Psychosomatics]. PMID- 9400154 TI - [Growing and doing yesterday and today. The disabled also can harvest]. PMID- 9400155 TI - [Alzheimer's dementia--a challenge to patients, family, physicians and society]. PMID- 9400156 TI - [Psychological aspects of the diagnostic process]. PMID- 9400157 TI - [Psychotherapeutic care of family and patients]. PMID- 9400158 TI - [Milieu therapy--a successful start]. PMID- 9400159 TI - [Alzheimer dementia: new perspectives in drug therapy]. PMID- 9400160 TI - [Laparoscopic "gastric banding" intervention for the treatment of pathologic obesity]. PMID- 9400161 TI - [The situation of skin tumors in Germany]. PMID- 9400162 TI - [Patients with skin diseases were made scapegoats of the health care system]. PMID- 9400163 TI - [Quality of life as an important condition in therapy--is it only a fad in medicine? Leaving decisions to the patients]. PMID- 9400164 TI - [Death--an everyday occurrence]. PMID- 9400165 TI - [Listening as an art and hard work. Why don't you ever listen to me?]. PMID- 9400166 TI - [When the Internet grows tiresome]. PMID- 9400167 TI - [Psychosomatics--2]. PMID- 9400168 TI - [Moving and touching to the end]. PMID- 9400170 TI - [What to do about the patients' sexual needs?]. PMID- 9400169 TI - [Sexuality of psychiatric patients. Neither nuns nor sex monsters]. PMID- 9400171 TI - [Nurses from a hundred cultures with common problems]. PMID- 9400173 TI - [A thought journey]. PMID- 9400172 TI - [Report from the Council of National Representatives, the legislative organ of the International Council of Nurses. How to achieve a fair representation of all the countries?]. PMID- 9400174 TI - [What is the role of the occupational nurse? "We play the role of catalysts"]. PMID- 9400175 TI - [Congress 1997: impressions. Health, a challenge to be shared]. PMID- 9400176 TI - [Nursing education in the year 2000. Towards clinical teaching]. PMID- 9400177 TI - [Visit by Madeleine Leininger, June 1997. "In Switzerland transcultural nursing is indispensible"]. PMID- 9400178 TI - [Improving teaching with clinical research]. PMID- 9400179 TI - [The importance of prevention]. PMID- 9400180 TI - [Collaboration in a project of prevention. The risk of falling in the elderly]. PMID- 9400181 TI - [... an unexpected change of contract]. PMID- 9400182 TI - [Three months colic. Your tips and experiences]. PMID- 9400183 TI - [A place at the feed bowl]. PMID- 9400184 TI - [Interdisciplinary cooperation by the 3 basic outpatient services. Each for himself and all together]. PMID- 9400185 TI - [Evolving nursing concepts and standards. Concrete measures for quality assurance]. PMID- 9400186 TI - [Quality in the treatment of dialysis patients]. PMID- 9400187 TI - [Time spaces]. PMID- 9400188 TI - [Support in psychosocial areas of life--but how and when?]. PMID- 9400189 TI - [The elbow ... the turning point]. PMID- 9400190 TI - [Challenges for the future]. PMID- 9400191 TI - [A concept of interdisciplinary treatment for patients with nutrition disorders. The obsession with being thin]. PMID- 9400192 TI - [Reform of the teaching system and implications for nursing. Toward a better professional education]. PMID- 9400193 TI - Mental health care. PMID- 9400194 TI - Behind the bare statistics. AB - The findings of the Mental Health Care stress survey will come as no surprise to nurses working on the front-line. But the statistics tell only half the story. PMID- 9400195 TI - Boateng gets down to business. AB - With his background in law and race relations, Paul Boateng was a surprise appointment as junior health minister. But he has quickly mastered his brief. PMID- 9400196 TI - Pathway to employment. PMID- 9400197 TI - Stress in mental health nursing: findings from the Mental Health Care survey. AB - Organisational and administrative concerns topped the list of stressors reported by a national sample of hospital and community mental health nurses. Seven of the ten top stressors were listed by both groups. Community nurses also listed inadequate service provision and lack of time to plan treatment. Hospital nurses were most stressed by inadequacy of staffing cover in potentially dangerous situations and low morale at work. Jerome Carson et al believe the findings indicate an urgent need for change at senior NHS management and administration level. PMID- 9400198 TI - Management strategies to tackle stress in mental health nursing. AB - Increased workloads, under-staffing, job insecurity and continuing, rapid organisational change have all been identified as major sources of stress among mental health nurses. So too has the increasing intensity of work, with more highly disturbed and potentially violent or suicidal patients. But, BEN THOMAS argues, trusts themselves could do much more by tackling poor management practice and introducing effective policies and procedures to deal with stress at both organisational and individual levels. PMID- 9400199 TI - Pressures and rewards of working in community mental health teams. AB - Community mental health teams (CMHTs) are widely regarded as the mainstay of community services, bringing health and social services professionals together to target people with severe mental health problems. Heather Harper and Edana Minghella report the findings of a recent survey looking at the pressures and rewards of working in these community-based, multi-disciplinary teams. PMID- 9400200 TI - Sharing medication information with patients. AB - Prompted by its user members, the Salford adult mental health services clinical audit group decided to review the information on medication available to clients. Here Mick Renoden, a service user on the clinical audit group with a 22 year history of contact with mental health services, describes his own experiences with clinicians and medication. Then Jeff Withington reports the results of a survey of users' views, which revealed a strong demand for information about medication that could be easily understood and trusted, and led to the production of a range of leaflets. PMID- 9400202 TI - Fighting fire with fire. AB - Post-traumatic stress disorder (PTSD) is a relatively recent diagnostic category, but it has long been recognised that trauma can produce persistent psychological sequelae. Paul Wheble describes how he successfully used cognitive behavioural therapy to help Alec, a fire-fighter, come to terms with an horrific accident 16 years previously. PMID- 9400201 TI - Epilepsy, learning disability and anti-convulsant drug status. AB - Epilepsy is frequently associated with learning disability, and can add considerably to the difficulties this client group experiences. However behavioural and daily living difficulties can be exacerbated by the seemingly common practice of polypharmacy, as this small-scale study shows. Barry Coughlan calls for further, detailed research into the appropriate administration of anti convulsant drugs to children and adults with learning disabilities and epilepsy. PMID- 9400203 TI - Door to opportunity. PMID- 9400204 TI - UK nurses have no right to feel complacent about the recent revelations that people with learning disabilities. PMID- 9400205 TI - Telling rights from wrong. AB - Is it right to medicate patients without their knowledge and consent? Or to tell an adult woman what she can eat? The UKCC is producing new guidance specifically to help mental health and learning disabilities nurses with the daily dilemmas of clinical practice. PMID- 9400206 TI - Bribery and co-operation. AB - NHS trusts are still looking for ways to maintain intensive continuing care in the community for people with severe mental illness. The US model of assertive community treatment (ACT) may be the answer. PMID- 9400207 TI - One hell of a trip. AB - In 1989 the uprising against the Ceausescu regime blew the lid off the hidden horrors of Romania's mental hospitals. Aid poured in from western agencies and charities. Eight years on, with attention switched elsewhere, how much has changed? PMID- 9400208 TI - Two tales of Mabel. PMID- 9400209 TI - Pulling together: multi-disciplinary training for mental health nursing. AB - In 1995 the Sainsbury Centre for Mental Health commissioned a major review of all specialist mental health training. The current, largely uni-disciplinary approach to training was felt to be failing to equip professionals with the necessary skills for today's multi-disciplinary, integrated, community-based service, where users and their carers expect an equal partnership and sharing of information. Kevin Gournay and Susannah Strong outline the findings and recommendations of the review, and its implications for mental health nurse education, which make a case for its separation from the rest of the nursing profession. PMID- 9400210 TI - Multi-disciplinary audit and the mental health nurse. AB - Research advances knowledge; audit is a way of ensuring that this new knowledge is applied to practice. Uni-disciplinary audit is well-established; multi disciplinary audit presents other challenges. Jonathan Ash outlines and illustrates how nurses working in a multi-disciplinary mental health team can initiate, design and use audit to improve the quality of the service, and to ensure it meets users' wants and needs. PMID- 9400211 TI - How do you identify people with severe mental illness in rural communities? AB - Mental health trusts serving scattered rural communities face particular challenges when attempting to identify and meet unmet need. A number of social, geographical and practical factors are involved. Kim Kirkwood and David Peck describe one such survey, carried out by the Highland Communities NHS Trust, the failure of which raises a number of important issues for practitioners and service planners. PMID- 9400212 TI - Love and friendship. AB - People with learning disabilities need to know about sexual health issues, and HIV/AIDS in particular. But how to convey complex and sensitive information in a clear and simple written format? Joelle Brogan shows it can be done. PMID- 9400213 TI - Disability discrimination. PMID- 9400214 TI - Finding a home. PMID- 9400215 TI - Learning together. PMID- 9400216 TI - Nurses promote breast health. PMID- 9400217 TI - The nurse's role in promoting breast cancer screening. AB - Early detection offers women the best chance of finding breast cancer early when it is most treatable. Both the ACS and the NCI now recommend mammography and CBE for women ages 40 and older. These organizations also recommend monthly BSE as a positive health practice. Nurses can play an important role in promoting screening by: (a) teaching women about screening guidelines, the benefits and limitations of screening, and risk factors for breast cancer and (b) helping women to reduce or eliminate barriers to screening. PMID- 9400218 TI - The diagnosis of breast abnormalities: nursing implications. PMID- 9400219 TI - Navigating the maze of breast cancer treatment. PMID- 9400220 TI - The New World of survivorship and rehabilitation. PMID- 9400221 TI - Breast cancer as a family disease. PMID- 9400222 TI - The impact of breast cancer on sexuality. PMID- 9400223 TI - Breast cancer and women partnering with women. PMID- 9400224 TI - Lesbian couples and cancer. PMID- 9400225 TI - Culturally competent nursing care related to breast cancer. PMID- 9400226 TI - Fear of recurrence: what you should know. PMID- 9400227 TI - But what about work? PMID- 9400228 TI - Spirituality and the nurse. PMID- 9400229 TI - Finding joy and freedom in the challenge of cancer. PMID- 9400230 TI - Down a twisted path. PMID- 9400231 TI - Internet breast cancer related resources. PMID- 9400232 TI - Driver's license: a model for future nursing regulation. PMID- 9400233 TI - A quest for independent living: a profile of Tim Kolb. PMID- 9400234 TI - Emphasis on employment. PMID- 9400235 TI - Female genital mutilation: the penultimate abuse experience. PMID- 9400236 TI - Visit from Japan. PMID- 9400237 TI - Oklahoma Nurses Association position statement unlicensed assistive personnel. PMID- 9400239 TI - One student's perspective. PMID- 9400238 TI - Prenatal weight gain and birth weight among Oklahoma mothers. PMID- 9400240 TI - A hospital's view of recovering nurses. PMID- 9400241 TI - Oklahoma Board of Nursing creates plan for regulation of unlicensed assistive personnel. PMID- 9400243 TI - AHCPR and private groups partner on pilot evidence reports. Agency for Health Care Policy and Research. PMID- 9400242 TI - Community partnership brings a response to health care concerns. PMID- 9400244 TI - Family Medical Leave Act. PMID- 9400245 TI - Political action committees: the work of ANA-PAC and ONA-PAC. PMID- 9400246 TI - Advanced practice nurses see rules and regulations approved by physicians. PMID- 9400247 TI - Smoking in Oklahoma: the 7-year trend. PMID- 9400248 TI - Oklahoma pregnancy risk assessment monitoring system. PMID- 9400249 TI - SWOSU nursing students working to increase awareness of the RN leaving the bedside. PMID- 9400250 TI - Every patient deserves a nurse every patient deserves you. PMID- 9400251 TI - A "caring" heart. PMID- 9400252 TI - "Tasks". PMID- 9400253 TI - Roles, functions of Oregon's clinical nurse specialists. PMID- 9400254 TI - ONA assisted suicide guidelines. PMID- 9400255 TI - ONA provides guidance on nurses' dilemma. PMID- 9400256 TI - Summary of ONA's top legislative priorities. PMID- 9400257 TI - NIOSH alerts health care workers to risk of developing latex allergy. PMID- 9400258 TI - Would you be ready to design your health care benefits? PMID- 9400259 TI - Antibiotic resistance: nursing implications for the '90s. PMID- 9400260 TI - ONA promotes image of nursing. PMID- 9400262 TI - [From the bed onto the operating table. The transfer of patients from their bed to the operating table]. AB - Perioperative nursing is still a much neglected area among the various fields of direct nursing care. This investigation was carried out within a framework of reevaluation and actualization of this area of care. The results show that in particular because of the working conditions of perioperative nursing high standards of professionally both with respect to details of care as well as to recognition of the overall situation of a patient are required. PMID- 9400261 TI - [Head outside--feet within. How do patients from differing cultures experience the German health care delivery system?]. PMID- 9400263 TI - [Linguistic confusion about nursing standards. A literature study]. AB - This paper takes a close look at nursing publications that are concerned with the theoretical knowledge of "standards". The central question is: "What are the intentions and characteristics to which standards are linked in nursing literature?" Diverse definitions and synonyms of standards are discussed as well. There is a considerable irregularity in the terminology of standards and similar terms in the investigated literature. Therefore, the term "standard" is treated by the author as a general one that embraces many definitions and has to be differentiated for a closer approach. Thus, four different kinds of features have been developed in the course of the analytic work: "intentions", "range of validity", "contents" and "structure". These four areas form a basis for a more definite description of the various kinds of standards. Based on her findings, the author give recommendations on how the actual linguistic confusion can be effectively dealt with. PMID- 9400264 TI - [Patterns of independent activities of daily living in the frail and disabled elderly: correlations with health and a socio-ecological context]. AB - The following contribution is focussing on independence in everyday life of 626 frail and disabled elderly people. The impact of medical and social factors on independence as well as different patterns of independence are examined. Results show that--besides medical factors--ecological living conditions (e.g. housing) strongly influence independence. Furthermore, four different patterns of independence with a high heterogenity in objective and subjective aspects of living conditions are analyzed. Finally, conclusions regarding nursing science and nursing practice are discussed. PMID- 9400265 TI - [Normative dimensions of nursing practice--the ethical relevance of the body]. AB - A combination of preliminary considerations concerning a theory of action and a philosophy of science illustrates the determining influence of the scientific preconception of the subject matter, and of the approach which is presupposed by this preconception, on the normative orientation of nursing practice. The specific physical nearness involved in nursing practice ("body to body") holds problems with regard to an appropriate theoretical frame of reference and corresponding practical convictions. This background provides the context for a concluding critical examination of several representative nursing theories with regard to their implicit, normative premises. PMID- 9400266 TI - [Communication in the hospital. A qualitative study of the communication behavior within the nursing service between the wards of a psychiatric department of a general hospital]. AB - In this project the behaviour of communication was examined of the nursing staff of a psychiatric clinic of a university hospital in Germany dealing with the staff of the other units. For this project qualitative methods of research with characteristics of the "Grounded Theory" have been used. The results show among others that communication barriers with regard to a cooperative communication process have existed in the past and are still showing some influence on todays' nursing staff. The interviewed nurses intend to realize a more cooperative communication and interaction on and between the units. A critical reflection of the "nursing conference", the "job rotation" or the "coaching of the team sessions by a neutral presenter" are examples of recommendations to improve and support the development of the communication process among the nurses. PMID- 9400267 TI - [Production of a classification system for nursing science for the library of Bremen University. A contribution to the development of nursing as a science]. AB - Nursing has taken important steps towards becoming a scientific discipline in Germany. The systematic registration and filing of the available literature in libraries needs to be enhanced in order to consolidate scientific infrastructures which can support activities in research, theory and international integration. In the course of teaching training studies in nursing science, a project was initiated to develop a classification system for nursing science literature at the University Library in Bremen. This article describes the genesis of the project, the basic conditions which were taken into account, the background and the developmental steps of the classification. PMID- 9400268 TI - [5 ambulances arrive too late]. PMID- 9400269 TI - [Narcotic abuse treatment--in contact with the mentally ill patient]. PMID- 9400270 TI - [Dr Jekyll and Mr. Hyde in the nursing home]. PMID- 9400271 TI - [Quality in the acute stage]. PMID- 9400272 TI - [Shallow network of ombudsman health visitors]. PMID- 9400273 TI - [At one's own responsibility]. PMID- 9400274 TI - [Rehabilitation in brain injury--the goal is independent activities]. PMID- 9400275 TI - [FS 25 (Professional Association 25) is dissolved]. PMID- 9400276 TI - [WHO--both private and public sector have a responsibility]. PMID- 9400277 TI - [Falck--lack of control]. PMID- 9400278 TI - [Velje district wants to ensure control with Falck]. PMID- 9400279 TI - [Relationship between profession and salary]. PMID- 9400280 TI - [Russia--neighbor in need]. PMID- 9400281 TI - [Russia--team nursing and strange therapies]. PMID- 9400282 TI - [Russia--the sick person in the East]. PMID- 9400283 TI - [Russia--outbreak of infectious diseases]. PMID- 9400284 TI - [Russia--a health system from the turn of the century]. PMID- 9400285 TI - [Russia--Finnish and Russian nurses cross borders]. PMID- 9400286 TI - [Russia--an indomitable ability to survive. Interview by Susanne Bloch Kjeldsen]. PMID- 9400287 TI - [Russia--new nurses for the new Russia]. PMID- 9400288 TI - [First Russian nurses' organization]. PMID- 9400289 TI - [Executive Board--education connection does not balance]. PMID- 9400290 TI - [Fast forward]. PMID- 9400291 TI - [Alcohol--personnel should intervene]. PMID- 9400292 TI - [Education--nursing theory: more freedom for patients]. PMID- 9400293 TI - [Early retirement--occupational ability only theoretical]. PMID- 9400294 TI - [Consultants should counsel on AMS]. PMID- 9400295 TI - [Russia--the good circle]. PMID- 9400296 TI - [Russia--exemplary home care]. PMID- 9400297 TI - [Russia--daily struggle to provide medicines]. PMID- 9400298 TI - [Russia--motivation has severe conditions]. PMID- 9400299 TI - [Russia--hope for Russian health]. PMID- 9400300 TI - [Organization--introduction to the debate--daring to act]. PMID- 9400301 TI - [Organization--watch out--the others are more dangerous!. Interview by Helle Broberg Nielsen]. PMID- 9400302 TI - [Organization--"Now we do it!". Interview by Helle Broberg Nielsen]. PMID- 9400303 TI - [Organization--troika management--an obsolete construction?. Interview by Helle Broberg Nielsen]. PMID- 9400304 TI - [Organization--ombudsman course: new wage forms take time. Interview by Soren Palsbo]. PMID- 9400305 TI - [Organization--ombudsman course: it is a change in system. Interview by Soren Palsbo]. PMID- 9400306 TI - [Organization--ombudsman course: ombudsman fully in front. Interview by Soren Palsbo]. PMID- 9400307 TI - [Organization--ombudsman course: an opportunity for the profession. Interview by Soren Palsbo]. PMID- 9400308 TI - [The little therapist]. PMID- 9400309 TI - [Quality assurance--we don't allow pressure sores here]. PMID- 9400310 TI - [Competence gap]. PMID- 9400312 TI - [Viewpoint from the medical technician. Interview by Marit Fonn]. PMID- 9400311 TI - [Hospital administrator challenges "the system". Interview by Kjelle Arne Bakke]. PMID- 9400313 TI - [I had a uniform and nameplate]. PMID- 9400314 TI - [Closeup: Gerd Fadnes, coordinator for reception of the sexually abused in Bergen emergency department. Supportive contact. Interview by Marit Fonn]. PMID- 9400315 TI - [Work environment--report use of overtime to Industrial Inspection. Interview by Tone Kristiansen]. PMID- 9400316 TI - [My workplace: home treatment team for HIV/AIDS patients, Uleval Hospital, Oslo- persons one gets to like. Interview by Kari Anne Aase]. PMID- 9400317 TI - [From times past--nurses' status in society]. PMID- 9400318 TI - [A split in the AF (Joint Academicians' Organization) could benefit NSF (Norwegian Nurses' Association). Interview by Kjell Arne Bakke]. PMID- 9400319 TI - [Sunnaas--State takes over TRS Center (Training and Counseling Center). Interview by Kari Anne Aase]. PMID- 9400320 TI - [Great pursuit of TSS (Telemark Central Hospital). Interview by Tone Kristiansen]. PMID- 9400321 TI - [National budget--a proposal ... and and explanation]. PMID- 9400322 TI - [Chronic lung disease--morning nap gives breathing a respite. Interview by Kjell Arne Bakke]. PMID- 9400323 TI - [France--vast differences between North and South]. PMID- 9400325 TI - [Overcoming crisis--a coping model with stress on feelings]. PMID- 9400326 TI - [Consumer participation--in focus groups patients are included in counseling]. PMID- 9400324 TI - [WHO--different ages for women and men]. PMID- 9400327 TI - [Competence--increased number of courses provide better professional development]. PMID- 9400328 TI - [Is behaviorism dangerous for nursing?]. PMID- 9400329 TI - The legacy of epidemiology in the Department of Social and Preventive Medicine. A commemoration of the sesquicentennial of the State University of New York at Buffalo School of Medicine and Biomedical Sciences. PMID- 9400330 TI - Observations on the history of epidemiology in western New York, 1843-1960. AB - Epidemiology has a rich tradition in western New York State, beginning with the classic study by Austin Flint of a waterborne typhoid fever outbreak in North Boston in 1843. Other important investigations included the study of the Buffalo poliomyelitis epidemic of 1912, by Wade Hampton Frost, which provided a comprehensive characterization of the epidemiology of the disease, and the first case-control study of cigarette smoking and lung cancer, by Morton L. Levin et al., conducted at the Roswell Park Memorial Cancer Institute in the 1940s. Other studies carried out before 1960 and included in the review deal with additional typhoid fever outbreaks, tuberculosis, breast cancer, and coronary heart disease. PMID- 9400331 TI - Current problems that are likely to affect the future of epidemiology. AB - The current discussion focuses on criticism as a positive force for improving epidemiologic practice through periodic reexamination of the basic approach to the discipline and the strategy for meeting the future educational needs of students and practicing epidemiologists. The types of epidemiologic research conducted and the settings within which the research will be conducted are also discussed. Epidemiology can be expected to play a major role in new areas of research that are created by changes in the medical care system and the development of large data systems associated with these approaches to health care delivery. This paper also discusses the growing threat to data access, the problems of communicating epidemiologic research findings to the public through the media, and the expanding interface between epidemiologic research and the legal system. The role of epidemiologic organizations in helping to shape the discipline's response to these issues and the opportunities these issues or problems present for improving epidemiologic research are also discussed. PMID- 9400332 TI - Toward an integrated approach to molecular epidemiology. AB - The emergence of "molecular epidemiology" as a scientific approach within the fields of epidemiology and toxicology has led to spirited discussion within the biomedical community, particularly in the area of cancer research. At scientific meetings and in peer-reviewed journals, numerous issues have been raised not only with regard to the practice of molecular epidemiology, but also with regard to its role in traditional epidemiology, toxicology, and risk assessment. Furthermore, the utility of information gleaned from such studies and the implications for public health have been the subject of considerable debate. Conceptual differences in how one views the function of epidemiologic and laboratory research may be reflected in discussions on the merits of molecular epidemiology. This commentary reviews some of the prevailing attitudes toward molecular epidemiology, with the goal of identifying areas of concern and suggesting means of achieving harmonization. The need for cross-training of epidemiologists and laboratory scientists is discussed, and suggestions are made for building successful collaborative relations across disciplines. PMID- 9400333 TI - Body mass index and mortality in a general population sample of men and women. The Buffalo Health Study. AB - The objective of this research was to investigate the long-term relation between body mass index (BMI) and mortality from all causes and from specific causes in the general population. A 29-year follow-up study was conducted in a random sample of white men (n = 611) and women (n = 697) aged 20-96 years who were residents of Buffalo, New York, in 1960. At baseline, height and weight were determined by self-report. BMI was calculated as weight (kg)/height (m2). During the follow-up period, 295 (48.3 percent) men and 281 (40.3 percent) women died. With the Cox proportional hazards model and adjustment for age, education, and cigarette smoking, a significant linear association was found between BMI and all cause mortality in men less than age 65 years at baseline (relative risk (RR) = 1.06, 95 percent confidence interval 1.02-1.09), but not in women (RR = 1.02, 95 percent confidence interval 0.99-1.05). In men age 65 years and older, the relation was quadratic in form (p = 0.02), with the lowest risks appearing in the BMI range of 23-27. BMI was most strongly related to cardiovascular disease (CVD) and coronary heart disease mortality in women and younger men. No such associations were observed in older men. BMI was not related to an increased risk of death from non-CVD or cancer in either sex. These findings illustrate the importance of BMI as a risk factor for CVD and coronary heart disease mortality in certain gender-age groups and indicate that the majority of the impact of BMI on overall mortality is due to the strong relation between relative weight and these specific causes of death. PMID- 9400334 TI - Lactation history and breast cancer risk. AB - Lifetime lactation in relation to breast cancer risk was examined in a case control study in two counties in western New York. Cases were women age 40 years and over with incident, primary, histologically confirmed breast cancer. Controls were age- and county-frequency matched, selected from New York State driver's license records (for those less than age 65 years) and from Health Care Finance Administration Records (for those age 65 or more). Included were women with at least one livebirth (253 premenopausal and 367 postmenopausal cases and 266 premenopausal and 427 postmenopausal controls). Breast cancer risk was very weakly associated with long duration of lactation among premenopausal women; the odds ratio for at least 20 months lifetime lactation was 0.50 (95% confidence interval 0.21-1.12). Among postmenopausal women, the protective effect of lactation was restricted to women with first lactation before age 25 years (odds ratio = 0.67, 95% confidence interval 0.46-0.95). However, age at first birth was highly correlated with age at first lactation. Neither insufficient milk as a reason for not breastfeeding nor having received medication to stop milk flow was associated with increased risk. These findings are in accordance with accumulating evidence that lactation may have a weak protective effect on breast cancer risk. PMID- 9400335 TI - Oxidative stress and lung function. AB - It has been suggested that lung function can be altered by both free radical and oxidant exposure, while antioxidant vitamin intake is positively related to lung function. However, the information on the relation of blood levels of oxidants and antioxidants to lung function is sparse. The present cross-sectional study, conducted from September 1995 to May 1996, analyzes the association between lung function measured as forced expiratory volume in 1 second (FEV1) with 1) levels of thiobarbituric acid-reactive substances in plasma (p-TBARS) and in low and very low density lipoprotein cholesterol (LDL cholesterol/VLDL cholesterol-TBARS) as indicators of lipid peroxidation and 2) compounds with antioxidant activity, erythrocytic glutathione, plasma glutathione peroxidase, trolox equivalent antioxidant capacity, and serum bilirubin, which may protect against lipid peroxidation. The analysis was carried out in 132 nonsmoking subjects aged 37-73 years who were randomly selected from the residents of Erie and Niagara counties, New York. FEV1 in percent of the predicted value (FEV1%) was negatively and statistical significantly associated with p-TBARS (r = -0.19). A negative association with borderline statistical significance was observed between FEV1% with low density lipoprotein cholesterol/very low density lipoprotein cholesterol TBARS (r = -0.16) and glutathione (r = -0.16), while FEV1% was positively related to serum bilirubin (r = 0.15). Participants in the lowest quartile of FEV1% showed significantly higher levels of p-TBARS (p = 0.02) and lower levels of bilirubin (p = 0.04) than did those in the highest quartile. Our results suggest that increased lipid peroxidation is associated with pulmonary airway narrowing in the general population. PMID- 9400336 TI - Consumption of contaminated sport fish from Lake Ontario and time-to-pregnancy. New York State Angler Cohort. AB - Sport fish from the Great Lakes are contaminated with halogenated organics, heavy metals, and pesticides, thus serving as a route of exposure for fish-consuming populations. These contaminants are recognized reproductive toxicants in animals; few human studies are available. The purpose of this study was to assess consumption of contaminated fish in relation to time-to-pregnancy (TTP) among women in the New York State Angler Cohort. In 1993, structured telephone interviews were conducted with 2,445 of 2,977 (82%) female cohort members aged 18 40 years who stated upon enrollment in the cohort in 1991 that they were considering pregnancy over the next 3 years. Among the 1,234 women who reported being pregnant, 874 (71%) had a known TTP and comprise the study sample. After descriptive analyses, log transformations of the number of years of fish consumption (duration) and TTP were performed and entered into multiple regression models that also included other covariates. Duration of fish consumption and maternal age accounted for only a small percentage of the explained variance in TTP (R2 = 0.005), even after the analysis was restricted to women who reported eating fish (R2 = 0.006). All beta coefficients were positive. These preliminary findings do not support an adverse effect of contaminated fish consumption on TTP. PMID- 9400337 TI - Consumption of PCB-contaminated freshwater fish and shortened menstrual cycle length. AB - Highly contaminated Lake Ontario sport fish represent an important human dietary exposure to polychlorinated biphenyls (PCBs) and other toxic contaminants that may disrupt endocrine pathways. New York State Angler Cohort women interviewed by telephone in 1993 provided menstrual cycle length (n = 2,223). Fish consumption at cohort enrollment in 1991 was categorized by duration and frequency and was used to calculate a PCB exposure index. Multiple regression analyses identified significant cycle length reductions with consumption of more than one fish meal per month (1.11 days) and moderate/high estimated PCB index (-1.03 days). Women who consumed contaminated fish for 7 years or more also had shorter cycles (-0.63 days). PMID- 9400338 TI - Physical activity, obesity, and diabetes in pregnancy. AB - Gestational diabetes mellitus (GDM) is the most common medical complication of pregnancy. Women with GDM are at elevated for numerous maternal health complications, and their infants are at elevated risk for death and morbidity. Management of GDM has traditionally been through diet and close monitoring of glucose levels, with initiation of insulin therapy when diet alone fails to maintain euglycemia. Recently, however, it has been suggested that alternative treatment modalities, such as exercise, may overcome a peripheral resistance to insulin, thus preventing GDM or controlling hyperglycemia in women with GDM. In this study, conducted from October 1995 to July 1996, the authors used a population-based birth registry to determine whether exercise has a preventive role in the development of GDM in women living in central New York State. They used contingency tables and chi-square statistics to examine bivariate differences among maternal and demographic variables and the occurrence of GDM. When stratified by prepregnancy body mass index category, exercise was associated with reduced rates of GDM only among women with a body mass index greater than 33 (odds ratio = 1.9, 95% confidence interval 1.2-3.1). The effect of exercise in obese women was further complicated by insurance status. When the data were stratified by insurance status, it appeared that women of higher socioeconomic status who were obese and did not exercise were at a significantly elevated risk of GDM compared with their counterparts of lower socioeconomic status. The results of this study suggest that for some women exercise may play a role in reducing the risk that they will develop GDM during pregnancy. PMID- 9400339 TI - Gender differences in the relation between depressive symptoms and alcohol problems: a longitudinal perspective. AB - Longitudinal relations between depressive symptoms and alcohol problems have been examined infrequently in community-based studies, and gender-specific findings to date appear to be inconclusive. Study hypotheses were that depressive symptoms predicted subsequent alcohol problems for females, whereas alcohol problems predicted subsequent depressive symptoms for males. The authors examined these relations in a random sample of household adults (aged 19 years or more) from Erie County, New York, assessed in 1986, 1989, and 1993 (n = 1,306). Measures of alcohol problems (in the previous year) incorporated an alcohol abuse/dependence diagnosis and a heavy alcohol use index. The Center for Epidemiologic Studies Depression Scale was used to assess depressive symptoms over a 1-month time frame. Comprehensive logistic regression models incorporated prior depressive symptoms, prior alcohol problems and sociodemographic variables (age, race, education, marital status, employment, total family income, and number of children living at home). For females, depressive symptoms predicted subsequent alcohol problems over 3 years (odds ratio = 3.04, 95% confidence interval 1.35 6.80, p < 0.01) and 4 years (odds ratio = 2.42, 95% confidence interval 1.14 5.12, p < 0.05), but not for 7 years. There was no evidence to support the hypothesis for males. This study clarifies and extends prior investigations of relations between these two prevalent mental health problems in a community-based sample. PMID- 9400340 TI - Test-retest reliability of the cognitive lifetime drinking history. AB - A new measure of lifetime alcohol consumption, the Cognitive Lifetime Drinking History (CLDH) uses beverage-specific questions on drink sizes and assesses drinking patterns to enhance recall. Two methods of establishing drinking intervals were examined: 1) floating--the respondent's report of when drinking changed, and 2) fixed--defined in terms of decades. Test-retest reliability for lifetime ounces of alcohol consumed and times intoxicated in lifetime estimated at visits 1 week or more apart was assessed in postmyocardial infarction patients (n = 81) and controls (n = 138) who had had at least 12 drinks in a year during their lifetimes. No significant differences in estimates of lifetime ounces of alcohol or times intoxicated were observed. Spearman's r ranged between 0.85 and 0.92 for the floating and fixed versions of the CLDH administered at a single visit and between 0.74 and 0.85 for the floating or fixed administered at both visits. Time between visits did not influence correlations. Intervals reported on the floating CLDH were comparable for postmyocardial infarction patients and controls. It took approximately 5 minutes longer to administer the floating CLDH than the fixed CLDH. Findings support use of the CLDH for case-control studies and suggest that the floating and fixed versions would yield comparable results. PMID- 9400341 TI - Cognitive aspects of recalling and reporting health-related events: Papanicolaou smears, clinical breast examinations, and mammograms. AB - This paper reports an examination of cognitive processes used by 178 women aged 50 years and older in retrieving information about the frequency with which they received Papanicolaou smears, mammograms, and clinical breast examinations. Women were selected from a health maintenance organization in which they had been enrolled for at least 5 1/2 years. The literature suggested that reporting of regular events such as these kinds of tests is likely to be based on schemas, which is an estimation technique in which events are reported in a format with generic content. Thus, if the procedure is believed to occur annually, the respondent will report receiving five tests in 5 years. The study attempted to evaluate whether use of episodic recall, in which respondents are forced to report individual events, would be more accurate than reports based on estimation using a schema format. The results indicated that most of the errors occurred in Papanicolaou smear reporting, which is consistent with the literature, and that the fewest errors occurred with mammograms. Regardless of the questionnaire format, respondents persisted in using schemas based on the date of annual physical examination. Most reporting errors occurred because the interval between examinations was estimated incorrectly. PMID- 9400342 TI - Clinical efficacy of methylphenidate in conduct disorder with and without attention deficit hyperactivity disorder. AB - BACKGROUND: Stimulants are not considered appropriate for the treatment of children with conduct disorders (CDs). The postulated differences in stimulant effect between children with attention deficit hyperactivity disorder (ADHD) and CD led to the hypothesis that methylphenidate hydrochloride, which is effective in ADHD, would not significantly improve symptoms of CD. METHODS: We randomly assigned 84 children with CD, between the ages of 6 and 15 years, to receive methylphenidate hydrochloride (up to 60 mg/d) or placebo for 5 weeks. Behavior was evaluated by parent, teacher, and clinician reports and by direct classroom observations. Two thirds of the children also met criteria for ADHD. RESULTS: Contrary to prediction, ratings of antisocial behaviors specific to CD were significantly reduced by methylphenidate treatment. The magnitude of methylphenidate effect indicated meaningful clinical benefit. Partialling out severity of ADHD did not alter the significant superiority of methylphenidate on CD ratings specifically (P < .001). CONCLUSIONS: Methylphenidate has short-term positive effects on children and adolescents with CD. Key aspects of antisocial adjustment appear to be treatment responsive. This effect was independent of severity of the children's initial ADHD symptoms. PMID- 9400343 TI - Fluoxetine and impulsive aggressive behavior in personality-disordered subjects. AB - BACKGROUND: Evidence of an inverse relationship between central serotonergic (serotonin [5-hydroxytryptamine]) system function and impulsive aggressive behavior has been accumulating for more than 2 decades. If so, pharmacological enhancement of serotonin activity should be expected to reduce impulsive aggressive behavior in subjects in whom this behavior is prominent. METHODS: A double-blind, placebo-controlled trial of the selective serotonin-uptake inhibitor fluoxetine hydrochloride was conducted in 40 nonmajor-depressed, nonbipolar or schizophrenic, DSM-III-R personality-disordered individuals with current histories of impulsive aggressive behavior and irritability. Measures included the Overt Aggression Scale-Modified for Outpatients, Clinical Global Impression Rating of Improvement, and several secondary measures of aggression, depression, and anxiety. RESULTS: Fluoxetine, but not placebo, treatment resulted in a sustained reduction in scores on the Irritability and Aggression subscales of the Overt Aggression Scale-Modified for Outpatients that was first apparent during months 2 and 3 of treatment, respectively. Fluoxetine was superior to placebo in the proportion of "responders" on the Clinical Global Impression Rating of Improvement: first at the end of month 1, and then finally demonstrating a sustained drug-placebo difference from the end of month 2 through the end of month 3 of treatment. These results were not influenced by secondary measures of depression, anxiety, or alcohol use. CONCLUSION: Fluoxetine treatment has an antiaggressive effect on impulsive aggressive individuals with DSM-III-R personality disorder. PMID- 9400344 TI - Dopamine receptor transcript expression in striatum and prefrontal and occipital cortex. Focal abnormalities in orbitofrontal cortex in schizophrenia. AB - BACKGROUND: The identification of novel subtypes of the dopamine receptors has renewed interest in the involvement of dopaminergic mechanisms in schizophrenia. We determined the expression of transcripts encoding the dopamine receptors in the brains of schizophrenic patients. METHODS: The levels of the messenger RNA molecules encoding the 5 dopamine receptors were quantified in postmortem brain samples from 16 schizophrenic patients and 9 control subjects. Samples from multiple regions of the prefrontal cortex, primary visual cortex, and striatum were subjected to in situ hybridization followed by quantitative image analysis. RESULTS: Expression of dopamine receptor transcripts did not differ between schizophrenic patients and controls in striatum or visual cortex. Dramatic decreases of dopamine receptor transcripts were found in the prefrontal cortex, but these changes were restricted to the D3 and D4 receptors, and localized to Brodmann area 11 (orbitofrontal cortex). CONCLUSIONS: Cortical dopaminergic neurotransmission may be disrupted in schizophrenia at the level of receptor expression. There appears to be a focal abnormality of D3 and D4 messenger RNA expression in the prefrontal cortex, with down-regulation of both, consistent with prefrontal cortical hypodopaminergia in schizophrenia. PMID- 9400346 TI - Structural magnetic resonance imaging abnormalities in men with severe chronic schizophrenia and an early age at clinical onset. AB - BACKGROUND: Early age at onset of schizophrenia often signifies a more severe form of the illness. However, the relationship between age at onset and brain abnormalities has not been established. We assessed temporal-limbic morphometry in severely ill men with chronic schizophrenia who had a relatively early onset of illness and examined the relationships among regional brain volumes, clinical symptoms, and age at illness onset. METHOD: Temporal lobe, superior temporal gyrus, hippocampus, temporal horn, lateral ventricles, third ventricle, and frontoparietal volumes were measured on magnetic resonance imaging data from 56 schizophrenic men (mean [SD] age at illness onset, 16.6 [4.2] years) recruited from a state hospital and 52 age- and range-matched healthy control men. RESULTS: Patients had significantly smaller gray matter volumes in the temporal lobe, superior temporal gyrus, and frontoparietal regions; smaller temporal lobe white matter volumes; and larger cerebrospinal fluid volumes for temporal lobe sulci and the 3 ventricular measures. There were no group differences in hippocampal volumes. Psychotic symptom subscores from the Brief Psychiatric Rating Scale were selectively correlated with smaller left posterior superior temporal gyrus gray matter volumes. None of the brain measurements were significantly correlated with age at illness onset. CONCLUSIONS: Data from this unique sample of severely ill schizophrenic men emphasize a pattern of structural abnormalities involving the cortex, but not the hippocampus, in schizophrenia. Furthermore, these data support theories suggesting that superior temporal gyrus abnormalities contribute selectively to psychotic symptoms and that the extent of structural abnormalities is unrelated to age of clinical symptom onset. PMID- 9400345 TI - The New York High-Risk Project. Prevalence and comorbidity of axis I disorders in offspring of schizophrenic parents at 25-year follow-up. AB - BACKGROUND: The New York High-Risk Project is a study of offspring of patients with schizophrenia (HRSz group) or affective illness (HRAff group) and psychiatrically normal parents (NC group) observed prospectively from childhood to adulthood. We herein present lifetime prevalence and comorbidity rates of Axis I disorders in subjects and their siblings from sample A of the project. METHODS: Schedule for Affective Disorders and Schizophrenia-Lifetime Version interviews conducted with the offspring in adulthood were used to obtain diagnoses of Axis I disorders. RESULTS: Schizophrenia and unspecified psychoses occurred only in the HRSz group. However, schizoaffective and psychotic affective disorders occurred equally in the HRSz and HRAff groups. Total rates of psychosis in these groups were significantly higher than in the NC group. All groups had similar rates of nonpsychotic affective and substance abuse disorders. The HRAff group, however, had significantly more total affective illness than the NC group and tended to have more anxiety disorders than the other groups. Comorbidity rates in the HRSz and HRAff groups were nearly twice those of the NC group. CONCLUSIONS: The familial liabilities to schizophrenia and affective disorders show specificities and commonalities, differing markedly from each other in their expression of some disorders and sharing others. Patterns of comorbidity are generally, although not entirely, similar to these liabilities. PMID- 9400347 TI - Neuroendocrine response to 5-hydroxy-L-tryptophan in prepubertal children at high risk of major depressive disorder. AB - BACKGROUND: Altered serotonergic function has been observed in prepubertal children and adults with an acute episode of major depressive disorder (MDD). However, it is not known whether these alterations are present prior to the onset of MDD. METHODS: A serotonergic precursor, 5-hydroxy-L-tryptophan (L-5HTP) (oxitriptan) (0.8 mg/kg), was administered through an indwelling catheter to 36 children at high risk of MDD (with high family loading for MDD), 31 children with MDD, and 23 low-risk normal controls (with low family loading for mood disorders and no history of psychopathology). Blood samples for cortisol, prolactin (PRL), and growth hormone were obtained every 15 minutes for 180 minutes, beginning 30 minutes before L-5HTP infusion. RESULTS: Children at high risk of MDD and children with MDD had similar hormonal responses following L-5HTP infusion. After controlling for baseline values, both groups secreted significantly less cortisol and more PRL than did the low-risk normal controls, with the PRL finding being limited to girls. There were no between-group differences in baseline cortisol, PRL, or growth hormone secretion measures. CONCLUSIONS: Before the onset of affective illness, high-risk children had the same pattern of neuroendocrine response to the L-5HTP challenge as did children with MDD. These results extend earlier findings of altered serotonergic regulation in association with early onset depression and indicate that these alterations may represent a trait marker for depression in children. PMID- 9400348 TI - What is the 'right' statistical measure of twin concordance (or diagnostic reliability and validity)? PMID- 9400349 TI - Another view on the 'right' statistical measure of twin concordance. PMID- 9400350 TI - The safety profile of naltrexone in the treatment of alcoholism. Results from a multicenter usage study. The Naltrexone Usage Study Group. AB - BACKGROUND: Naltrexone hydrochloride is the first medication approved in the United States for the treatment of alcohol dependence in almost 50 years. This study was designed to collect safety data in a setting that reflected the expected clinical use of naltrexone. METHODS: This was a 12-week, nonrandomized, open-label usage study conducted in 40 alcoholism treatment centers throughout the United States, including free-standing alcoholism treatment programs, university clinics, Veterans Administration hospitals, and office-based primary care practices. Eligible patients were assigned, at the investigators' discretion, to a naltrexone treatment group or to a reference group that did not receive study medication. At study entry, patients must have been abstinent from alcohol for 1 to 6 weeks and enrolled in a psychosocial treatment program for alcoholism. Patients often underrepresented in controlled clinical trials, including women and patients with comorbid medical and psychiatric illness, were eligible. Patients with polysubstance abuse or infection with the human immunodeficiency virus were not excluded. RESULTS: Of 865 patients enrolled, 570 received naltrexone and 295 were in a reference group. The most common new-onset adverse clinical events in the naltrexone group were nausea (9.8%) and headache (6.6%). Naltrexone was discontinued in 15.0% of patients because of adverse events, most frequently nausea. The results of liver function tests in the naltrexone group were similar to those in the reference group. No death occurred during the study. CONCLUSIONS: This is the largest study to date describing the safety of naltrexone in a heterogeneous population of persons with alcoholism. No new safety concerns were identified. PMID- 9400351 TI - Hypercortisolism associated with social subordinance or social isolation among wild baboons. AB - BACKGROUND: The phenomena of basal hypercortisolism and of dexamethasone resistance have long intrigued biological psychiatrists, and much is still unknown as to the causes and consequences of such adrenocortical hyperactivity in various neuropsychiatric disorders. We have analyzed basal cortisol concentrations and adrenocortical responsiveness to dexamethasone in a population of wild baboons living in a national park in Kenya. We tested whether social subordinance in a primate is associated with dexamethasone resistance. Furthermore, we examined whether individual differences in adrenocortical measurements were predicted by the extent of social affiliation in these animals. METHODS: Seventy yellow baboons (Papio cynocephalus) were anesthetized and injected with 5 mg of dexamethasone; the cortisol response was monitored for 6 hours. The animals were of both sexes in a range of ages and had known ranks in the dominance hierarchies within their troops. Extensive behavioral data were available for a subset of 12 adult males who were anesthetized under circumstances that also allowed for the determination of basal cortisol concentrations. RESULTS: The socially subordinate baboons were less responsive to dexamethasone than were the dominant ones; as one manifestation of this, postdexamethasone cortisol values were more than 3 times higher in the dozen lowest-ranking animals compared with the dozen highest. In addition, socially isolated males had elevated basal cortisol concentrations and showed a trend toward relative dexamethasone resistance. CONCLUSIONS: Our findings indicate that social status and degree of social affilitation can influence adrenocortical profiles; specifically, social subordinance or social isolation were associated in our study with hypercortisolism or feedback resistance. PMID- 9400352 TI - Autopsy-proven Alzheimer disease in a patient with dementia who retained musical skill in life. PMID- 9400353 TI - Nobel Prize in Physiology or Medicine for 1997 Awarded to Stanley B. Prusiner, MD. PMID- 9400354 TI - Abnormal expression of laminin beta 1 chain in skeletal muscle of adult-onset limb-girdle muscular dystrophy. AB - BACKGROUND: Laminin 2 is a major component of the basal lamina of skeletal muscle cells. It is a heterotrimer composed of 3 chains: merosin (laminin alpha 2 chain), beta 1, and gamma 1. Deficiency of merosin, with or without laminin beta 1 chain reduction, is associated with some forms of congenital muscular dystrophy. Deficient expression of laminin beta 1 chain is also associated with some cases of merosin-positive congenital muscular dystrophy. The expression of laminin 2 subunits has not been well studied in the skeletal muscle of limb girdle muscular dystrophy (LGMD), nor has much attention been given to the significance of reduction of individual laminin 2 subunits, such as beta 1. OBJECTIVES: To examine the expression of laminin 2 subunits in skeletal muscle in patients with LGMD and to define the clinical features of patients with LGMD who have abnormal expression of laminin 2 subunits. METHODS: We studied muscle biopsy specimens from 18 patients with LGMD using immunofluorescence with antibodies against dystrophin C-terminus, beta-dystroglycan, alpha-sarcoglycan, gamma sarcoglycan, and the laminin subunits merosin, beta 1, and gamma 1. Of the 18 biopsy specimens, 9 were available for electron microscopic examination of the muscle basement membrane. The clinical features associated with abnormal laminin beta 1 chain immunoreactivity were further described. RESULTS: Laminin beta 1 chain was either barely detectable or severely reduced in 3 cases of patients with LGMD in which the biopsy specimens showed normal staining with the other antibodies. Patients in all 3 cases had common clinical features consistent with a slowly progressive, adult-onset LGMD. Specimens from 2 of the 3 cases that were available for ultrastructural examination showed significant abnormalities of the muscle fiber basement membrane. CONCLUSIONS: Abnormal expression of laminin beta 1 chain without concomitant deficiency of alpha-sarcoglycan in skeletal muscle has not been previously described in LGMD. Reduced laminin beta 1 chain immunoreactivity may potentially serve as a marker for defining subsets of individuals with LGMD, in particular those with slowly progressive, adult-onset pelvifemoral presentation. The abnormality of muscle fiber basement membranes in specimens from cases that were available for ultrastructural study suggests that defects in the extracellular matrix may play a role in the pathogenesis of this subset of LGMD. PMID- 9400355 TI - Total quality improvement method for reduction of delays between emergency department admission and treatment of acute ischemic stroke. The National Institute of Neurological Disorders and Stroke rt-PA Stroke Study Group. AB - OBJECTIVE: To develop an approach for reducing time between emergency department (ED) admission and treatment in patients with acute ischemic stroke to meet the challenge of providing tissue plasminogen activator treatment within 180 minutes. DESIGN: An observational study. SETTING: Forty trial-affiliated hospitals, including 30 community hospitals. PARTICIPANTS: A total of 17,324 consecutive patients admitted to trial-affiliated hospital EDs within 24 hours of possible stroke, from January 1991 through October 1994. INTERVENTION: Appraisal of the process of triage, evaluation, diagnosis, and treatment by means of total quality improvement techniques in each hospital. Staff participating in the process identified sources of variation and modifications by flow charting the process. MAIN OUTCOME MEASURE: Time between ED admission and treatment with study medication. RESULTS: Total quality improvement methods identified hospital specific process improvements. Many improvements were administrative, requiring no additional resources. More than 50% of screened patients arrived too late to be treated. Only 1268 patients were admitted between 0 and 125 minutes from stroke onset with no other trial exclusion criteria; 48% were treated. Of 243 patients admitted between 126 and 170 minutes from stroke onset with no exclusion criteria, 4% were treated. Mean time from ED admission to treatment was similar in teaching and community hospitals. CONCLUSIONS: Total quality improvement methods identified ED-specific sources of process variability and reduced time between ED admission and treatment. Therefore, these methods should be considered in developing and monitoring emergent stroke treatment protocols. PMID- 9400356 TI - The quality-of-life effects of interferon beta-1b in multiple sclerosis. An extended Q-TWiST analysis. AB - BACKGROUND: A recombinant form of interferon beta-1b (Betaseron) was given Food and Drug Administration approval for use in the treatment of relapsing-remitting multiple sclerosis in 1993 based on a documented reduction in exacerbation rate. However, its effect on disease progression is less clear. It costs $11,000 per year and has documented adverse effects such as fatigue, feverlike symptoms, and depression. OBJECTIVES: To evaluate a recombinant form of interferon beta-1b in the treatment of relapsing-remitting multiple sclerosis and to discuss treatment trade-offs and comprehensive quality-of-life (QOL) outcomes. METHODS: We present a randomized evaluation of treatment with a recombinant form of interferon beta 1b in 79 patients with multiple sclerosis who participated in a random allocation lottery and were followed up for 12 months, during which data on QOL and clinical outcomes were collected. The data were analyzed using the Extended Quality Adjusted Time Without Symptoms and Toxicity (Q-TWiST) method, which evaluates treatment trade-offs by incorporating several QOL domains and patient preferences regarding these domains. RESULTS: Over the 12 months of follow-up, the case patients reported 10.6 months of quality-adjusted time, while the control patients reported 10.4 months of quality-adjusted time (P = .50). CONCLUSIONS: Thus, the first year of treatment with interferon beta-1b did not significantly improve or detract from QOL. Results are discussed in terms of acceptable trade offs depending on the nature of therapy. Future observational and clinical studies should incorporate measures of patient preference. PMID- 9400357 TI - A magnetic resonance imaging study of planum temporale asymmetry in men with developmental dyslexia. AB - BACKGROUND: Imaging studies have suggested anomalous anatomical asymmetries in language-related regions of the temporal and parietal lobes in individuals with developmental dyslexia. Autopsy studies have reported unusual symmetry of the planum temporale (PT) in patients with dyslexia. Methodological limitations characterize much of this literature, however. OBJECTIVE: To examine the size and asymmetry of the PT and its extension into the parietal lobe (planum parietale [PP]) in men with well-characterized, persistent dyslexia by using magnetic resonance imaging and 3-dimensional surface rendering techniques. METHODS: The brains of 16 right-handed dyslexic men aged 18 to 40 years and 14 matched control subjects were studied with magnetic resonance imaging. Most of these subjects were previously studied with positron emission tomography, which demonstrated functional abnormalities in temporal and parietal brain regions in the dyslexic group. The area of the PT was determined with the aid of 3-dimensional surface rendering techniques. The size of the PP was estimated by measuring the length of the posterior ascending ramus on 3 parasagittal slices. RESULTS: Approximately 70% to 80% of both groups showed equivalent leftward (left > right) asymmetries of the PT; approximately 50% to 60% showed equivalent rightward (right > left) asymmetries of the PP. These asymmetries showed equivalent moderate inverse correlations with each other in both groups. CONCLUSIONS: These results challenge the notion that anomalous asymmetry of the PT is strongly associated with developmental dyslexia. Given the heterogeneity of the dyslexic population, some subgroup of dyslexic individuals (i.e., those with developmental language disorders) may show unusual symmetry or reversed asymmetry in this region. However, anomalous asymmetry of the planum did not contribute to functional abnormalities demonstrated in these patients by positron emission tomography. PMID- 9400358 TI - Dichotic-listening performance and intracarotid injections of amobarbital in children and adolescents. Preoperative and postoperative comparisons. AB - BACKGROUND: Dichotic listening (DL) to consonant-vowel syllables is frequently used in clinical and experimental studies of brain laterality. However, the paradigm of consonant-vowel syllables has not been thoroughly validated through a comparison with injections of amobarbital sodium (Amytal). OBJECTIVE: To validate the DL test for hemisphere dominance preoperatively vs postoperatively with the results from intracarotid injections of amobarbital (i.e., the Wada test) in epileptic children and adolescents. DESIGN AND MAIN OUTCOME MEASURES: Patients were tested with DL preoperatively and at 6-month follow-up. Correct reports in the DL tests were entered in a stepwise discriminant analysis for calculation of correct classification of hemisphere dominance with the results from the injections of amobarbital as the grouping variable. Correct reports from the right and left ears on the consonant-vowel DL test were compared preoperatively and postoperatively, separated for the subjects with regard to language dominance in the left and right hemispheres. SETTING: The Department of Pediatrics, Ostra Hospital, University of Goteborg, Goteborg, Sweden. PATIENTS: Thirteen children and adolescents between the ages of 10 and 19 years, who were surgically treated for resistant epilepsy, were included in the study. The operated area corresponded with morphological changes and functional dysfunctions according to findings from computed tomography, magnetic resonance imaging, single photon emission computed tomography, and electroencephalography. RESULTS: The results of the Wada tests revealed that 10 subjects had left hemisphere language dominance, with 3 subjects having right hemisphere language dominance. All 3 subjects with right hemisphere language dominance showed a left ear advantage on the DL test preoperatively and postoperatively, with 8 and 7 of the 10 subjects with left hemisphere dominance showing a right ear advantage, preoperatively and postoperatively, respectively. However, according to discriminant analysis, knowledge of the DL performance led to a correct classification according to the Wada test results in 12 (92%) of the 13 subjects. CONCLUSIONS: A quantitative classification procedure like discriminant analysis may be more sensitive when predicting hemisphere speech dominance from DL data than a qualitative procedure based on the ear advantage dichotomy. The ear advantage dichotomy may actually introduce arbitrary left-right categories that do not correspond to the actual clustering of the data. PMID- 9400359 TI - Prose recall in dementia. A comparison of delay intervals. AB - OBJECTIVE: To explore one methodological variation, delay length, that may contribute to contradictory findings in the literature regarding the use of delayed recall in the detection of early-stage dementia of the Alzheimer type. DESIGN: Comparison of participants with dementia and without dementia on a prose recall task at both 10- and 30-minute delay intervals. SETTING: Washington University Alzheimer's Disease Research Center, St Louis, Mo. PARTICIPANTS: Participants with very mild dementia of the Alzheimer type (n = 136) and uncompromised elderly individuals (n = 197). MAIN OUTCOME MEASURES: Results of the Logical Memory subtest from the Wechsler Memory Scale with immediate recall and 10- and 30-minute delayed recall. RESULTS: Participants with dementia recalled significantly less material than elderly controls at both immediate and delayed recall (P < .001). Multiple regression analyses revealed that dementia classification failed to account for additional variance in the 30-minute delayed score beyond that which could be accounted for by the immediate score. A small but significant proportion of variance was accounted for in the 10-minute delayed score beyond that which could be accounted for by the immediate recall score. CONCLUSION: Delayed recall of a prose passage does not appear to enhance the differentiation of very mild dementia of the Alzheimer type from normal aging in a meaningful way, whether the recall delay is 10 or 30 minutes. PMID- 9400360 TI - Predictors of intracranial pathologic findings in patients who seek emergency care because of headache. AB - BACKGROUND: Clinical criteria to select patients with headache in whom structural diagnostic studies (computed tomography) have a high yield disclosing intracranial pathologic findings, independent of abnormal findings on neurologic examination, have not been defined. OBJECTIVE: To determine which clinical characteristics predict the presence of intracranial pathologic findings, independently of neurologic examination, in patients with headache. DESIGN: Case control, consecutive sample. SETTING: Major metropolitan trauma center emergency department. PATIENTS AND MATERIALS: Hospital records of 139 hospitalized and 329 randomly selected patients from 1720 nonhospitalized adult patients, consecutively evaluated for headache in the emergency department, were reviewed. Demographic data, clinical characteristics of the headache, results of neurologic and physical examinations, and diagnostic radiologic and laboratory results were correlated with final diagnosis and outcome at 6 months after emergency department visit. DATA ANALYSIS: Nonparametric statistical analysis. RESULTS: Intracranial pathologic findings were found in 18 (3.8%) of 468 patients. Acute onset and occipitonuchal location of headache, presence of associated symptoms, and patient age of 55 years or older were significantly associated with the finding of intracranial pathology, independently of the findings from neurologic examination. Abnormal findings on neurologic examination alone, whether focal or nonfocal, had a highly significant association and a positive predictive value for intracranial pathology of 39%. CONCLUSIONS: Abnormal results from neurologic examination are the best clinical parameters to predict structural intracranial pathology; however, in patients 55 years or older with headache of acute onset located in the occipitonuchal region that has associated symptoms, computed tomographic scan of the head is justified as part of their clinical evaluation independently of the findings of the neurologic examination. PMID- 9400361 TI - 'Complete' spinal cord injury does not block perceptual responses to genital self stimulation in women. AB - BACKGROUND: A priori hypothesis: vaginal and/or cervical self-stimulation will not produce perceptual responses in women with "complete" spinal cord injury (SCI) at or above the highest level of entry of the hypogastric nerves (T10-12) but will produce perceptual responses if SCI is below T-10. DESIGN: Women with complete SCI were assigned to a group with "upper" (T-10 and/or above) (n = 6) or "lower" (below T-10) (n = 10) SCI; uninjured women (n = 5) constituted a control group. Perceptual response to vaginal and/or cervical self-stimulation was quantified as magnitude of analgesia to calibrated finger compressive force. SETTING: Rutgers, The State University of New Jersey, Human Physiology Laboratory, College of Nursing, Newark. PARTICIPANTS: Consecutive samples of first 16 of 34 women with SCI who responded to nationwide advertisements, met inclusion criteria, and volunteered; control group was the first 5 respondents. INTERVENTION: Vaginal or cervical (cervix uteri) self-stimulation applied for 12 minutes, interspersed with non-stimulation periods, while measuring analgesia. MAIN OUTCOME MEASURE: Quantify analgesia magnitude to vaginal or cervical self stimulation. RESULTS: Significant analgesia was produced in the uninjured group and the group with lower SCI, supporting the hypothesis. Unexpectedly, significant analgesia was also produced in the group with upper SCI. Women in the group with upper SCI also experienced menstrual discomfort, awareness of vaginal and/or cervical stimulation per se, and orgasms. CONCLUSIONS: (1) Genitospinal visceral afferent pathways function in the women in the group with upper SCI, although unrecognized by the American Spinal Injury Association criteria, and/or (2) there exists a functional genital afferent pathway that bypasses the spinal cord and projects directly to the brain, which we propose to be via the vagus nerves. PMID- 9400362 TI - Volumetric magnetic resonance imaging. Clinical applications and contributions to the understanding of temporal lobe epilepsy. AB - In recent years, magnetic resonance imaging-based volumetric measurements of the amygdala and hippocampus have proved useful in the diagnosis and treatment of patients with temporal lobe epilepsy. This imaging modality allows amygdaloid and hippocampal volumes to be correlated with neurophysiological, neuropathological, and neuropsychological findings, surgical outcome, and clinical findings. We evaluated the technical and anatomical aspects underlying the successful use of the modality that were reported in previous studies. We also evaluated issues such as the sensitivity and specificity of volumetric magnetic resonance imaging, its use in bilateral temporal lobe epilepsy, and the debate concerning the sensitivity of qualitative visual analysis vs quantitative volumetric analysis of magnetic resonance images. Volumetric magnetic resonance imaging, when used in conjunction with video electroencephalographic monitoring, neuropsychological studies, and other neuroimaging studies, will enable patients with temporal lobe epilepsy to be treated in an appropriate, efficient, and cost-effective manner. PMID- 9400363 TI - Adult-onset Niemann-Pick type C disease. Clinical, biochemical, and genetic study. AB - BACKGROUND: Niemann-Pick type C disease is an autosomal recessive neurometabolic disorder of unknown origin mapped to chromosome 18q11-12 in most of the studied families. In contrast to the sphingomyelin lipidoses, in Niemann-Pick type C disease, fibroblasts are impaired in intracellular homeostatic responses to exogenous low-density lipoprotein (LDL) cholesterol. Biochemical heterogeneity of the disorder in relation to abnormal LDL processing is associated with various clinical presentations, but adult-onset Niemann-Pick type C disease is rare and has not been comprehensively characterized. OBJECTIVE: To describe clinical, biochemical, and genetic features of adult-onset Niemann-Pick type C disease in 3 siblings. DESIGN AND SETTING: Case series in a tertiary care center. PATIENTS: The 3 siblings manifested a variable combination of vertical supranuclear ophthalmoplegia, ataxia, and splenomegaly. Brain magnetic resonance imaging showed cerebellar atrophy; brainstem auditory evoked responses were unobtainable, and bone marrow examination disclosed typical foam cells. The patients were 20, 26, and 28 years old and belonged to a sibship of 13 born of consanguineous healthy parents. METHODS: Esterification of exogenous LDL cholesterol in cultured skin fibroblasts and filipin staining for free intracellular cholesterol. Polymerase chain reaction-based DNA linkage study using AC microsatellite markers D18S40, D18S44, D18S480, and D18S66. RESULTS: Fibroblasts of the 3 patients showed a 23% to 58% block in the induced cholesterol esterification after 4 1/2 hours and a mild to moderate accumulation of free cholesterol. DNA study demonstrated linkage to the major 18q11-12 Niemann-Pick type C locus and identified unaffected carriers. CONCLUSIONS: These results confirm the diagnosis of the least biochemically affected Niemann-Pick type C phenotype in this family with adult-onset disease and support a correlation between the mild laboratory and clinical findings in this age group. PMID- 9400364 TI - Four legs. Illusory reduplication of the lower limbs after bilateral parietal lobe damage. AB - OBJECTIVE: To report an unusual disorder of body schema and its neurologic and neuropsychological correlates. DESIGN AND METHODS: We describe a patient with a reduplicative phantom illusion of her lower limbs. Motor and sensory functions, as well as mental representation of body and space, were studied during the reduplication experience until its resolution. SETTING: Clinical neurology department in a primary care hospital. PATIENT: A 64-year-old, left-handed woman who experienced the uncontrollable and distressing feeling of having 4 legs, without delusional belief, after surgical removal of a right-predominant parasagittal parietal meningioma. This phenomenon spontaneously resolved after 2 weeks. INTERVENTION: None. MAIN OUTCOME MEASURES: Clinical neurologic examinations and standardized neuropsychological tests, with emphasis on tests assessing orientation to body parts, right-left discrimination, and mental orientation in space. RESULTS: The patient had severe weakness and proprioceptive sensory loss in both lower limbs. She had no disturbances of body schema knowledge but a striking impairment in tasks requiring mental orientation in space, particularly for right-left laterality discrimination. Resolution of the reduplication experience correlated with improvement in the affected spatial abilities, while motor, sensory, and other cognitive functioning did not significantly change. CONCLUSION: This patient's reduplicative phantom illusion might be related to the combination of the severe somatosensory loss with an underlying impaired mental representation of relative positions in space. PMID- 9400365 TI - Model of a quinary structure between Krebs TCA cycle enzymes: a model for the metabolon. AB - The enzymes which are responsible for catalyzing sequential reactions in several metabolic pathways have been proposed to be highly organized in supramolecular complexes termed metabolons. However, the in situ existence of these weak complexes is difficult to demonstrate because many of them are dissociated during isolation due to dilution effects. Consequently, the metabolon concept is subject to controversy. A model system consisting of genetically prepared bienzymatic fusion proteins has been used to immobilize sequential metabolic enzymes in close proximity and to demonstrate possible kinetic advantages of metabolons. These experiments use the sequential Krebs TCA cycle enzymes from yeast mitochondrial malate dehydrogenase (MDH), citrate synthase (CS), and aconitase (ACO). Using the porcine high-definition structures of these three enzymes, we have performed computer-modeling studies in order to understand how the molecules may interact. Among the thousands of docking orientations we have tried, one was found to respond to the structural and experimental constraints from the results obtained with the yeast fusion proteins. Interestingly, this quinary structure model shows substantial interacting surface areas with spatial and electrostatic complementarities which make the complex thermodynamically stable. This structure also contains an unbroken electrostatically favorable channel connecting the active sites of ACO and CS, as well as the one previously reported between CS and MDH active sites. Charged amino acids which could be involved in interactions stabilizing the complex have been identified. This model will be used as the basis for further experimental work on the structure of the Krebs TCA cycle metabolon. PMID- 9400366 TI - An early intermediate in the folding reaction of the B1 domain of protein G contains a native-like core. AB - The folding kinetics of a 57-residue IgG binding domain of streptococcal protein G has been studied under varying solvent conditions, using stopped-flow fluorescence methods. Although GB1 has been cited as an example of a protein that obeys a two-state folding mechanism, the following kinetic observations suggest the presence of an early folding intermediate. Under stabilizing conditions (low denaturant concentrations, especially in the presence of sodium sulfate), the kinetics of folding shows evidence of a major unresolved fluorescence change during the 1.5 ms dead time of the stopped-flow experiment (burst phase). Together with some curvature in the rate profile for the single observable folding phase, this provides clear evidence of the rapid formation of compact states with native-like fluorescence for the single tryptophan at position 43. In refolding experiments at increasing denaturant concentrations, the amplitude of the sub-millisecond phase decreases sharply and the corresponding slope (m value) is only about 30% lower than that of the equilibrium unfolding curve indicative of a pre-equilibrium transition involving cooperative unfolding of an ensemble of compact intermediates. The dependence on guanidine hydrochloride concentration of both rates and amplitudes (including the equilibrium transition) is described quantitatively by a sequential three-state mechanism, U [symbol: see text] I [symbol: see text] N, where an intermediate (I) in rapid equilibrium with the unfolded state (U) precedes the rate-limiting formation of the native state (N). A 66-residue fragment of GB1 with an N-terminal extension containing five apolar side chains exhibits three-state kinetic behavior virtually identical to that of the 57-residue fragment. This is consistent with the presence of a well-shielded native-like core excluding the N-terminal tail in the early folding intermediate and argues against a mechanism involving random hydrophobic collapse, which would predict a correlation between overall hydrophobicity and stability of compact states. PMID- 9400367 TI - Cysteine-scanning mutagenesis of helix IV and the adjoining loops in the lactose permease of Escherichia coli: Glu126 and Arg144 are essential. off. AB - Cys-scanning mutagenesis has been applied to the remaining 45 residues in lactose permease that have not been mutagenized previously (from Gln100 to Arg144 which comprise helix IV and adjoining loops). Of the 45 single-Cys mutants, 26 accumulate lactose to > 75% of the steady state observed with Cys-less permease, and 14 mutants exhibit lower but significant levels of accumulation (35-65% of Cys-less permease). Permease with Phe140-->Cys or Lys131-->Cys exhibits low activity (15-20% of Cys-less permease), while mutants Gly115-->Cys, Glu126-->Cys and Arg144-->Cys are completely unable to accumulate the dissacharide. However, Cys-less permease with Ala or Pro in place of Gly115 is highly active, and replacement of Lys131 or Phe140 with Cys in wild-type permease has a less deleterious effect on activity. In contrast, mutant Glu126-->Cys or Arg144-->Cys is inactive with respect to both uphill and downhill transport in either Cys-less or wild-type permease. Furthermore, mutants Glu126-->Ala or Gln and Arg144-->Ala or Gln are also inactive in both backgrounds, and activity is not rescued by double neutral replacements or inversion of the charged residues at these positions. Finally, a mutant with Lys in place of Arg144 accumulates lactose to about 25% of the steady state of wild-type, but at a slow rate. Replacement of Glu126 with Asp, in contrast, has relatively little effect on activity. None of the effects can be attributed to decreased expression of the mutants, as judged by immunoblot analysis. Although the activity of most of the single-Cys mutants is unaffected by N-ethylmaleimide, Cys replacement at three positions (Ala127, Val132, or Phe138) renders the permease highly sensitive to alkylation. The results indicate that the cytoplasmic loop between helices IV and V, where insertional mutagenesis has little effect on activity [McKenna, E., et al. (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 11954-11958], contains residues that play an important role in permease activity and that a carboxyl group at position 126 and a positive charge at position 144 are absolutely required. PMID- 9400368 TI - Crystal structure of phenylmethanesulfonyl fluoride-treated human chymase at 1.9 A. AB - The X-ray crystal structure of human chymase has been determined to 1.9 A resolution using molecular replacement methods. This first structure of human chymase is present as the Ser 195 ester of alpha-toluenesulfonic acid. The refined structure (Rcryst = 0.183) shows that the inhibitor phenyl moiety lies at the top of the major specificity pocket, S1, while the sulfur is covalently linked to Ser 195-O gamma. The sulfonyl oxygens interact with the oxyanion hole and with His 57-N delta 1. The presence of the inhibitor disturbs the usual gauche position of His 57 and forces it to the trans conformer. Though the primary binding pockets are similarly specific in chymase and chymotrypsin, examination of the extended substrate binding sites reveals the structural basis for chymase's greater discrimination in choosing substrates. The larger 30s loop and its proximity to the active site indicates that it contacts substrate residues C-terminal to the scissile bond. Modeling of substrate at the chymase active site suggests that binding energy may be gained by three main-chain hydrogen bonds provided by substrate residues P2' and P4' and that discriminating interactions with substrate side chains are also likely. The presence of Lys 40 in S1' of human chymase explains its preference for Asp/Glu at P1'. Moreover, the cationic nature of S1' provides a structural basis for human chymase's poor catalytic efficiency when angiotensin II is the substrate. PMID- 9400369 TI - Increased pressure induces sustained protein kinase C-independent herbimycin A sensitive activation of extracellular signal-related kinase 1/2 in the rabbit aorta in organ culture. AB - The 42- and 44-kD mitogen-activated protein kinases, also referred to as extracellular signal-related kinase (ERK) 2 and 1, respectively, may be transiently activated by stretching vascular smooth muscle cells (VSMCs). Using an organ culture model of rabbit aorta, we studied short- and long-term ERK1/2 activation by intraluminal pressure (150 mm Hg). Activation of ERK1/2 was biphasic: it reached a maximum (217.5 +/- 8.4% of control) 5 minutes after pressurizing and decreased to 120.7 +/- 5.1% of control after 2 hours. Furthermore, after 24 hours of pressurizing, ERK1/2 activity was as high (241.8 +/- 14.7% of control) as in the acute phase. Long-term pressure-induced ERK1/2 activation correlated with stimulation of tyrosine phosphorylation of proteins in the 125- to 140-kD range. Neither protein kinase C inhibitors (1 mumol/L staurosporine or 50 mumol/L bisindolylmaleimide-I) nor tyrosine kinase inhibitors (50 mumol/L tyrphostin A48 or 50 mumol/L genistein) affected pressure-induced ERK1/2 activation. However, the Src-family tyrosine kinase inhibitor herbimycin A (500 nmol/L) did reduce both 5-minute (by 92 +/- 8%) and 24-hour (by 63 +/- 7%) pressure-induced ERK1/2 activation. Thus, our results demonstrate a sustained activation of ERK1/2 and tyrosine kinases by intraluminal pressure in the arterial wall. Pressure-induced ERK1/2 activation is PKC independent and Src family tyrosine kinase dependent and possibly includes activation of extracellular matrix-associated tyrosine kinases. PMID- 9400370 TI - Activation of MAP kinase in vivo follows balloon overstretch injury of porcine coronary and carotid arteries. AB - Vascular restenosis involves contraction, proliferation, and remodeling of the arterial wall in response to overstretch injury. Mitogen-activated protein kinases (MAPKs) are implicated in both contraction and proliferation of vascular smooth muscle (VSM), and studies of porcine carotid arterial muscle strips have shown that mechanical stretch leads to the activation of the extracellular signal regulated kinase (ERK) family of MAPKs in vivo. We, therefore, analyzed the acute effect of mechanical overstretch injury on ERK-MAPK (herein referred to simply as MAPK) activity in porcine coronary and carotid arteries in vivo. Balloon angioplasty catheters were inflated to 6 atm three times over 5 minutes at a balloon-artery ratio of 1.2:1 in either porcine coronary or carotid arteries. The arteries were snap-frozen after angioplasty, and MAPK activity was measured. Angioplasty of the left anterior descending (LAD, n = 5), left circumflex (LCx, n = 5), and carotid (n = 5) arteries effected an increase in MAPK activity compared with the activity in uninstrumented right coronary arteries (RCAs) or carotid arteries from the same animals used for controls. Balloon angioplasty of carotid arteries led to an increase in MAPK activity that was 7.7-fold over the activity in control arteries and comparable to the activity in stretched carotid arterial muscle strips in vivo. The increase in coronary artery kinase activity on angioplasty was variable from animal to animal. The increase in MAPK activity over that in control arteries ranged from 4.5- to 31.7-fold (mean +/- SEM, 10.7 +/- 5.3) in the LAD and 1.8- to 31.3-fold (mean +/- SEM, 9.7 +/- 5.7) in the LCx. There were no apparent inherent differences in the levels of MAPK activity in the three different types of coronary arteries (RCA, LAD, and LCx) without instrumentation. MAPK activation occurs rapidly during angioplasty, suggesting that this kinase may play an early role in initiating the injury response in both porcine coronary and carotid arteries. MAPKs may be key enzymes targeted to treat or prevent restenosis. PMID- 9400371 TI - Transcriptional regulation of inducible nitric oxide synthase in cultured neonatal rat cardiac myocytes. AB - Previous work has demonstrated that inducible NO synthase (iNOS) can be expressed in cardiac myocytes. In this study, we investigated transcriptional regulation of the iNOS gene in these cells. Lipopolysaccharide (LPS) induced iNOS mRNA and protein in cultured neonatal rat cardiac myocytes. H-89, dexamethasone, herbimycin, genistein, staurosporine, or pyrrolidine dithiocarbamate (PDTC) attenuated the iNOS induction by LPS. Forskolin, interleukin (IL)-6, tumor necrosis factor (TNF)-alpha, or interferon (IFN)-gamma enhanced the LPS-induced iNOS expression. Combined stimulation of IL-6 and TNF-alpha also induced iNOS. The 5'-upstream sequence of the rat iNOS gene contains the nuclear factor-kappa B (NF-kappa B) site, CAAT box, IFN-gamma activation site (GAS), and IFN regulatory factor (IRF) site. DNase I footprinting assay revealed that the nuclear factors binding to these elements were increased by LPS exposure. Transient transfection assay suggested that these elements were indispensable for transcriptional regulation of the iNOS induction. Electrophoretic mobility shift assay revealed that LPS or TNF-alpha increased binding activity for the NF-kappa B site. A slower-migrating complex binding to the CAAT box gave rise after exposure to LPS or forskolin. Competition assay suggested that this slower-migrating complex consisted of a heterodimer between a member of CAAT box/enhancer binding (C/EBP) protein family and cAMP responsive element binding protein (CREB). LPS or IL-6 increased binding complexes for the IRF site, which was compatible with induction of IRF-1. LPS, IL-6, or IFN-gamma induced a novel binding complex for GAS, which also existed in the 5'-flanking region of the IRF-1 gene. These data suggest that (1) iNOS induction simultaneously requires both NF-kappa B activation and IRF-1 induction, and (2) the heterodimer between C/EBP and CREB has synergistic effects on the iNOS induction via the CAAT box. PMID- 9400372 TI - Molecular mechanisms of anoxia/reoxygenation-induced neutrophil adherence to cultured endothelial cells. AB - The objectives of this study were to (1) determine the time course of neutrophil adhesion to monolayers of human umbilical vein endothelial cells (HUVECs) that were exposed to 60 minutes of anoxia followed by 30 to 600 minutes of reoxygenation and (2) define the mechanisms responsible for both the early (minutes) and late (hours) hyperadhesivity of postanoxic HUVECs to human neutrophils. The results clearly demonstrate that anoxia/reoxygenation (A/R) leads to a biphasic increase in neutrophil adhesion to HUVECs, with peak responses occurring at 30 minutes (phase 1) and 240 minutes (phase 2) after reoxygenation. Oxypurinol and catalase inhibited phase-1 adhesion, suggesting a role for xanthine oxidase and H2O2. In comparison, platelet activating factor (PAF) contributed to both phases of neutrophil adhesion. Anti-intercellular adhesion molecule-1 (ICAM-1) and anti-P-selectin antibodies (monoclonal antibodies [mAbs]) attenuated phase-1 neutrophil adhesion, consistent with roles for constitutively expressed ICAM-1 and enhanced surface expression of preformed P-selectin. Phase-2 neutrophil adhesion was attenuated by an anti-E-selectin mAb, indicating a dominant role of this adhesion molecule in the late phase response. Pretreatment with actinomycin D and cycloheximide or with competing ds oligonucleotides containing the nuclear factor-kappa B or activator protein-1 cognate DNA sequences significantly attenuated phase-2 response, suggesting a role for de novo macromolecule synthesis. Surface expression of ICAM-1, P selectin, and E-selectin on HUVECs correlated with the phase-1 and -2 neutrophil adhesion responses. Collectively, these findings indicate that A/R elicits a two phase neutrophil-endothelial cell adhesion response that involves transcription independent and transcription-dependent surface expression of different endothelial cell adhesion molecules. PMID- 9400373 TI - Vascular endothelial growth factor increases the mitogenic response to fibroblast growth factor-2 in vascular smooth muscle cells in vivo via expression of fms like tyrosine kinase-1. AB - Vascular endothelial growth factor (VEGF) has traditionally been considered an endothelial cell-specific factor inducing angiogenesis and vascular permeability in vivo. In the present study, expression of VEGF and its receptors, fetal liver kinase-1 (flk-1) and fms-like tyrosine kinase-1 (flt-1), was examined in rat carotid arteries after balloon injury. Although VEGF and flk-1 were not detectable, high levels of flt-1 mRNA and protein were expressed by smooth muscle cells (SMCs) in the neointima, as demonstrated by en face in situ hybridization and Western blotting. Intimal SMC proliferation in chronically denuded rat carotid arteries was unaffected by intraluminal infusion of VEGF, whereas fibroblast growth factor (FGF)-2 increased the number of replicating SMCs 4-fold. Pretreatment with VEGF doubled the mitogenic response to infused FGF-2 by increasing SMC replication in deeper layers of the intima. VEGF increased the permeability of chronically denuded vessels to plasma proteins but had no effect on the uptake of locally infused biotinylated FGF-2. These findings demonstrate that vascular SMCs express functional flt-1 receptors after arterial injury and that VEGF has synergistic effects with FGF-2 on SMC proliferation. These effects are likely to be mediated by a VEGF-mediated increase in permeability as well as a direct interaction between the VEGF and FGF signaling pathways. PMID- 9400374 TI - Smooth muscle cells isolated from discrete compartments of the mature vascular media exhibit unique phenotypes and distinct growth capabilities. AB - Heterogeneity of smooth muscle cell (SMC) phenotype and function is rapidly emerging as an important concept. We have recently described that phenotypically distinct SMC subpopulations in bovine pulmonary arteries exhibit unique proliferative and matrix-producing responses to hypoxic pulmonary hypertension. To provide better understanding of the molecular mechanisms contributing to this phenomenon, experimental studies will require a reliable in vitro model. The purpose of the present study was first to determine if distinct SMC subpopulations, similar to those observed in vivo, could be selectively isolated from the mature arterial media, and then to evaluate whether select SMC subpopulations would exhibit heightened responses to growth-promoting stimuli and hypoxia. We were able to reproducibly isolate at least four phenotypically unique cell subpopulations from the inner, middle, and outer compartments of the arterial media. Differences in cell phenotype were demonstrated by morphological appearance and differential expression of muscle-specific proteins. The isolated cell subpopulations exhibited markedly different growth capabilities. Two SMC subpopulations grew slowly in 10% serum and were quiescent in plasma-based medium. The other two cell subpopulations, exhibiting nonmuscle characteristics, grew rapidly in 10% serum and proliferated in plasma-based medium and in response to hypoxia. Certain colonies of the nonmuscle-like cell subpopulations were found to grow autonomously under serum-deprived conditions and to secrete mitogenic factors. Our data, demonstrating that phenotypically distinct cells with enhanced growth potential exist within the normal arterial media, support the idea that these unique cells could contribute selectively to the pathogenesis of vascular disease. PMID- 9400375 TI - Mechanisms of action of troglitazone in the prevention of high glucose-induced migration and proliferation of cultured coronary smooth muscle cells. AB - Recent findings suggest that high glucose levels may promote atherosclerosis in coronary vascular smooth muscle cells (VSMCs). To explore the intracellular mechanisms of action by which troglitazone affects this process, we examined the effect of troglitazone on the migration and growth characteristics of cultured rabbit coronary VSMCs. Treatment with chronic high glucose medium (22.2 mmol/L) for 5 days increased VSMC migration by 92%, [3H]thymidine incorporation by 135%, and cell number by 32% compared with VSMCs treated with normal glucose (5.5 mmol/L glucose + 16.6 mmol/L mannose) medium. Trolitazone at 100 nmol/L and 1 mumol/L significantly suppressed high glucose-induced VSMC migration by 34% and 42%, respectively, the proliferative effect (as measured by cell number) by 17% and 27%, and [3H]thymidine incorporation by 45% and 60% (n = 6, P < .05). The high glucose-induced impairment of insulin-mediated [3H]deoxyglucose uptake was blocked by a protein kinase C (PKC) inhibitor (calphostin C, 1 mumol/L) and was also improved by troglitazone without any change in insulin receptor number and affinity. The high glucose-induced insulin-mediated increase in cell number and in [3H]thymidine incorporation was suppressed by troglitazone. Troglitazone (1 mumol/L) also suppressed high glucose-induced phospholipase D activation, elevation of the cytosolic NADH/NAD+ ratio (as measured by the cytosolic ratio of lactate/pyruvate), and membrane-bound PKC activation. Flow cytometric DNA histogram analysis of cell cycle stage showed that high glucose-induced increase in the percentage of cells in the S phase was suppressed by 1 mumol/L troglitazone. These findings suggest that PKC may be a link between impairment of insulin-mediated glucose uptake and the increase in migration and proliferation induced by high glucose levels and that troglitazone may be clinically useful for the treatment of high glucose-induced coronary atherosclerosis. PMID- 9400376 TI - Mevalonate-dependent inhibition of transendothelial migration and chemotaxis of human peripheral blood neutrophils by pravastatin. AB - Pravastatin, a hydrophilic inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase, has been reported to beneficially affect atherogenesis, plaque stability, and transient myocardial ischemia in significant coronary artery disease by influencing lipid metabolism and by intracellular signaling via mevalonate pathway products other than cholesterol. Leukocytes are implicated to play a pathophysiological role in these events. We were interested in finding out whether pravastatin could affect transendothelial migration (TEM), chemotaxis, and respiratory burst activity of the neutrophil ex vivo. In addition, effects on monocyte and T-lymphocyte chemotaxis were tested. For TEM assays, monolayers of human umbilical vein endothelial cells (HUVECs) were grown to confluence on polycarbonate filters bearing 5-microns pores in Transwell (Costar) culture plate inserts. Chemotaxis experiments were performed using modified Boyden chambers with cellulose nitrate micropore filters. Respiratory burst activity was measured fluorometrically. Treatment of neutrophils and monocytes with pravastatin at 2 to 200 mumol/L and 10 to 1000 mumol/L, respectively, significantly decreased chemotaxis triggered by fMet-Leu-Phe. This effect was abolished in the presence of mevalonic acid (500 mumol/L); no effect of pravastatin was seen on T lymphocyte chemotaxis triggered by interleukin-8. Preincubation of neutrophils with pravastatin (200 mumol/L) also resulted in a significant reduction in the number of neutrophils that transmigrated a tumor necrosis factor-stimulated or lipopolysaccharide-stimulated HUVEC monolayer. At none of the concentrations tested (2 pmol/L to 200 mumol/L) did pravastatin affect neutrophil respiratory burst activity. We conclude that pravastatin may alter monocyte chemotaxis and neutrophil-endothelial interactions in migratory responses at concentrations obtained in vivo with cholesterol-lowering doses. PMID- 9400377 TI - Angiotensin II induces apoptosis of human endothelial cells. Protective effect of nitric oxide. AB - Angiotensin II (Ang II) importantly contributes to the pathobiology of atherosclerosis. Since endothelial injury is a key event early in the pathogenesis of atherosclerosis, we tested the hypothesis that Ang II may injure endothelial cells by activation of cellular suicide pathways leading to apoptosis. Human umbilical venous endothelial cells (HUVECs) were incubated with increasing doses of Ang II for 18 hours. Apoptosis of HUVECs was measured by ELISA specific for histone-associated DNA fragments and confirmed by DNA laddering and nuclear staining. Ang II dose-dependently induced apoptosis of HUVECs. Simultaneous blockade of both the AT1 and AT2 receptor prevented Ang II induced apoptosis, whereas each individual receptor blocker alone was not effective. Selective agonistic stimulation of the AT2 receptor also dose dependently induced apoptosis. Ang II-mediated as well as selective AT2 receptor stimulation-mediated apoptosis was associated with the activation of caspase-3, a central downstream effector of the caspase cascade executing the cell death program. Specific inhibition of caspase-3 activity abrogated Ang II-induced apoptosis. In addition, the NO donors sodium nitroprusside and S nitrosopenicillamine completely inhibited Ang II-induced apoptosis and eliminated caspase-3 activity. Thus, Ang II induces apoptosis of HUVECs via activation of the caspase cascade, the central downstream effector arm executing the cell death program. NO completely abrogated Ang II-induced apoptosis by interfering with the activation of the caspase cascade. PMID- 9400378 TI - Nitric oxide-independent dilation of conductance coronary arteries to acetylcholine in conscious dogs. AB - NO and prostacyclin formation cannot entirely account for receptor-operated endothelium-dependent dilation of coronary vessels, since vasodilator responses are not completely suppressed by inhibitors of these agents. Therefore, we considered that another factor, such as an endothelium-derived hyperpolarizing factor described in vitro, may participate in NO- and prostacyclin-independent coronary dilator responses. In conscious instrumented dogs, intracoronary acetylcholine (ACh, 30.0 ng.kg-1.min-1) increased the external epicardial coronary diameter (CD) by 0.18 +/- 0.03 mm (from 3.44 +/- 0.11 mm) when increases in coronary blood flow (CBF) were prevented and increased the CD by 0.20 +/- 0.05 when CBF was allowed to increase. After the administration of intracoronary N omega-nitro-L-arginine methyl ester (L-NAME), CBF responses to ACh were abolished, but CD responses (0.23 +/- 0.05 from 3.22 +/- 0.09 mm) were maintained. Blockade of NO formation was confirmed by reduced CD baselines and blunted flow-dependent CD responses caused by adenosine and transient coronary artery occlusions after L-NAME administration. ACh-induced CD increases resistant to L-NAME and indomethacin were reduced after the administration of intracoronary quinacrine, an inhibitor of phospholipase A2, or proadifen, an inhibitor of cytochrome P-450. Quinacrine or proadifen alone (without L-NAME) did not alter CD responses to ACh, but L-NAME given after proadifen blunted ACh-induced increases in CD. The increases in CD caused by arachidonic acid given after L-NAME + indomethacin were antagonized by proadifen but not altered by quinacrine. Thus, a cytochrome P-450 metabolite of arachidonic acid accounts for L-NAME-resistant and indomethacin-resistant dilation of large epicardial coronary arteries to ACh. Conversely, NO formation is the dominant mechanism of ACh-induced dilation after blockade of the cytochrome P-450 pathway. PMID- 9400379 TI - Venous myogenic tone and its regulation through K+ channels depends on chronic intravascular pressure. AB - In this study, we compared the level of myogenic tone and its negative-feedback control through specific K+ channels in two types of human veins (saphenous [SV] and cephalic [CV] veins), which experience different ranges of pressure in vivo. We also investigated whether an experimental model of increased venous pressure in rats exposed to head-up tilt for 2 weeks produced changes similar to those observed in the human veins. Cylindrical vein segments were cannulated, their diameters were measured, and the intraluminal pressure was set at different levels (2 to 30 mm Hg) in vitro. Acetylcholine test showed that during the regular harvesting process 76% of the human SVs exposed for coronary bypass grafts had no functional endothelium. We found significant myogenic tone in the human SV, where the in vivo pressure is high, but it was not present in the human CV, where the in vivo pressure is low. The nonspecific K+ channel antagonist, tetraethylammonium (TEA), decreased the diameter of the human SV but not the CV. Iberiotoxin and 4-aminopyridine, blockers of the Ca(2+)-sensitive (KCa) and voltage-gated K+ (KV) channels, also decreased the diameter of the human SV by 10.2 +/- 4.8% and 19.5 +/- 4.7%, respectively. In the rat SV, significant myogenic tone was found, but TEA had no effect, even after 2 weeks of in vivo pressure increase in the hindlimb by head-up tilt. We conclude that (1) an increased venous myogenic tone correlates with higher chronic intraluminal pressure loads, (2) KCa and KV channels counterregulate the myogenic tone in human, but not in rat, saphenous vein, (3) the counterregulatory effect is more effective at high than at low intraluminal in vitro pressure levels, and (4) its development is probably a long-term process. PMID- 9400380 TI - Increased myogenic tone and diminished responsiveness to ATP-sensitive K+ channel openers in cerebral arteries from diabetic rats. AB - Diabetes mellitus has profound adverse effects on vascular and, in particular, endothelial function. Although pressure-induced constriction ("myogenic tone") is a major contributor to the regulation of blood flow, little is known about the effects of diabetes on this response. Diabetes has been shown to diminish the dilation of cerebral arteries to synthetic ATP-sensitive K+ (KATP) channel openers. In this study, we explored the effects of diabetes induced in rats by streptozotocin on cerebral artery (250 to 300 microns) myogenic tone and on vasodilations to the synthetic KATP channel openers pinacidil and levcromakalim. Elevation of intravascular pressure caused a graded membrane potential depolarization and constriction, which was greater in arteries from diabetic rats compared with normal rats (at 60 mm Hg, 5 mV more depolarized and 22 microns more constricted). Pressurized arteries (at 60 mm Hg) from diabetic rats were 5- to 15 fold less sensitive to pinacidil and levcromakalim than were control arteries (EC50 values for pinacidil and levcromakalim were 1.4 and 0.6 mumol/L, respectively, in diabetic animals and 0.3 and 0.04, respectively, in control animals; P < .05). Removal of the endothelium or addition of a NO synthase inhibitor, NG-nitro-L-arginine (LNNA), in control arteries decreased the sensitivity to KATP channel openers and depolarized and constricted control arteries to levels similar to those observed in arteries from diabetic animals. Sodium nitroprusside caused a membrane potential hyperpolarization and enhanced the response to pinacidil in arteries from diabetic animals. Removal of the endothelium or LNNA had little effect on the apparent KATP channel opener sensitivity, the membrane potential, and pressure-induced constrictions of arteries from diabetic animals. The results are consistent with the hypothesis that this type of diabetes leads to a decrease in tonic NO release from the endothelium, which in turn causes membrane potential depolarization and vasoconstriction, resulting in a diminished response to KATP channel openers. PMID- 9400381 TI - Targeted ablation of the murine alpha-tropomyosin gene. AB - We created a mouse that lacks a functional alpha-tropomyosin gene using gene targeting in embryonic stem cells and blastocyst-mediated transgenesis. Homozygous alpha-tropomyosin "knockout" mice die between embryonic day 9.5 and 13.5 and lack alpha-tropomyosin mRNA. Heterozygous alpha-tropomyosin knockout mice have approximately 50% as much cardiac alpha-tropomyosin mRNA as wild-type littermates but similar alpha-tropomyosin protein levels. Cardiac gross morphology, histology, and function (assessed by working heart preparations) of heterozygous alpha-tropomyosin knockout and wild-type mice were indistinguishable. Mechanical performance of skinned papillary muscle strips derived from mutant and wild-type hearts also revealed no differences. We conclude that haploinsufficiency of the alpha-tropomyosin gene produces little or no change in cardiac function or structure, whereas total alpha-tropomyosin deficiency is incompatible with life. These findings imply that in heterozygotes there is a regulatory mechanism that maintains the level of myofibrillar tropomyosin despite the reduction in alpha-tropomyosin mRNA. PMID- 9400382 TI - Lipopolysaccharide depresses cardiac contractility and beta-adrenergic contractile response by decreasing myofilament response to Ca2+ in cardiac myocytes. AB - Lipopolysaccharide (LPS) plays a key role in the pathogenesis of sepsis. Cardiac function and the inotropic response to beta-adrenergic stimulation are impaired in sepsis. We hypothesized that LPS, in clinically relevant levels (1 ng/mL), directly depresses contractility and beta-adrenergic responses in cardiac myocytes. Cardiac myocytes were isolated from the left ventricle of adult rabbits using digestive enzymes (collagenase and protease). We depyrogenated the enzymes (LPS contamination lowered from 100 to 300 ng/mL to < 0.7 ng/mL) to minimize development of LPS tolerance during cell isolation. After 6 hours of incubation with 1 ng/mL LPS, there was a decrease in the extent of active cell shortening with no change in Ca2+ transients (measured with indo 1 fluorescence), indicating decreased myofilament responsiveness to Ca2+. This was related to NO pathways, since cGMP (a second messenger of NO) increased in cardiac myocytes and LPS effects were completely reversed with a 1 mmol/L NG-monomethyl-L-arginine (L NMMA, a NO synthase inhibitor). LPS did not alter the intracellular Ca2+ response to beta-adrenergic stimulation with isoproterenol but attenuated the contractile response (maximal cell shortening, 15.5 +/- 1.0% versus 23.3 +/- 1.1% in control myocytes; P < .001). LPS attenuation of the contractile response to isoproterenol was restored completely by L-NMMA and almost completely restored (to 86% of the control response) by an inhibitor of cGMP-dependent protein kinase. We conclude that LPS depresses cardiac contractility and the contractile response to beta adrenergic stimulation by a NO-cGMP-mediated decrease in myofilament responsiveness to Ca2+. The direct effects of low levels of LPS on cardiac myocytes may contribute to cardiac depression and hemodynamic decompensation during sepsis. PMID- 9400383 TI - beta-Arrestin1 knockout mice appear normal but demonstrate altered cardiac responses to beta-adrenergic stimulation. AB - beta-Arrestin1 knockout mice were studied to define the physiological role of beta-arrestin1 in the regulation of G protein-coupled receptors. beta-Arrestin1 is thought to be involved in the desensitization of many G protein-associated cell surface receptors, particularly beta-adrenergic receptors. Homozygous knockout mice are overtly normal. Resting cardiovascular parameters modulated by beta-adrenergic receptors such as heart rate, blood pressure, and left ventricular ejection fraction are not changed. However, homozygous mutants are more sensitive to beta-receptor agonist-stimulated increases in ejection fraction, consistent with a role of beta-arrestin1 in beta-adrenergic receptor desensitization. We conclude that beta-arrestin1 is important for in vivo G protein-coupled receptor desensitization and that this aspect of desensitization represents a mechanism for fine-tuning responses. However, beta-arrestin1 does not appear to be required for development or for other essential biological functions. PMID- 9400384 TI - Experimental diabetes is associated with functional activation of protein kinase C epsilon and phosphorylation of troponin I in the heart, which are prevented by angiotensin II receptor blockade. AB - A cardiomyopathy that is characterized by an impairment in diastolic relaxation and a loss of calcium sensitivity of the isolated myofibril has been described in chronic diabetic animals and humans. To explore a possible role for protein kinase C (PKC)-mediated phosphorylation of myofibrillar proteins in this process, we characterized the subcellular distribution of the major PKC isoforms seen in the adult heart in cardiocytes isolated from diabetic rats and determined patterns of phosphorylation of the major regulatory proteins, including troponin I (TnI). Rats were made diabetic with a single injection of streptozotocin, and myocardiocytes were isolated and studied 3 to 4 weeks later. In nondiabetic animals, 76% of the PKC epsilon isoform was located in the cytosol and 24% was particulate, whereas in diabetic animals, 55% was cytosolic and 45% was particulate (P < .05). PKC delta, the other major PKC isoform seen in adult cardiocytes, did not show a change in subcellular localization. In parallel, TnI phosphorylation was increased 5-fold in cardiocytes isolated from the hearts of diabetic animals relative to control animals (P < .01). The change in PKC epsilon distribution and in TnI phosphorylation in diabetic animals was completely prevented by rendering the animals euglycemic with insulin or by concomitant treatment with a specific angiotensin II type-1 receptor (AT1) antagonist. Since PKC phosphorylation of TnI has been associated with a loss of calcium sensitivity of intact myofibrils, these data suggest that angiotensin II receptor-mediated activation of PKC may play a role in the contractile dysfunction seen in chronic diabetes. PMID- 9400385 TI - Low efficiency of Ca2+ entry through the Na(+)-Ca2+ exchanger as trigger for Ca2+ release from the sarcoplasmic reticulum. A comparison between L-type Ca2+ current and reverse-mode Na(+)-Ca2+ exchange. AB - It has been proposed that Ca2+ entry through the Na(+)-Ca2+ exchanger can contribute significantly to the trigger for Ca2+ release from the sarcoplasmic reticulum (SR). We have compared the characteristics of Ca2+ release triggered by reverse-mode Na(+)-Ca2+ exchange and by L-type Ca2+ current (ICaL) during depolarizing steps in single guinea pig ventricular myocytes (whole-cell voltage clamp, fluo 3 and fura-red as [Ca2+]i indicators, 36 +/- 1 degrees C, K(+)-based pipette solution with 20 mmol/L [Na+]). Conditioning pulses to +60 mV ensured comparable Ca2+ loading of the SR. In the presence of ICaL, [Ca2+]i transients typically have an early and rapid rising phase reflecting Ca2+ release, which has a bell-shaped voltage dependence with a peak at +10 mV. With Ca2+ entry through Na(+)-Ca2+ exchange only (20 mumol/L nisoldipine), Ca2+ release flux from the SR is decreased and directly related to the amplitude of the depolarizing step. Ca2+ release is preceded by a significant delay (81 +/- 21 ms at +20 mV, 24 +/- 4 ms at +70 mV) related to Ca2+ entry through the exchanger. Triggered release interrupts Ca2+ entry, as evidenced by reversal of the exchanger current. At potentials positive to +40 mV, Ca2+ influx through Na(+)-Ca2+ exchange, calculated from the outward exchange current, reaches magnitudes comparable to ICaL, but Ca2+ release due to reverse-mode Na(+)-Ca2+ exchange still has a significant delay. We calculated trigger efficiency as the ratio between the maximal rate of Ca2+ release and the Ca2+ influx preceding this release; efficiency of reverse-mode Na(+)-Ca2+ exchange is approximately four times less than that of ICaL. With both ICaL and reverse-mode Na(+)-Ca2+ exchange present, Ca2+ release is triggered by ICaL, and a contribution of reverse-mode Na(+)-Ca2+ exchange to the trigger could not be detected at potentials below +60 mV. These characteristics of reverse-mode Na(+)-Ca2+ exchange predict that its role as a trigger for Ca2+ release during the action potential is likely to be negligible. PMID- 9400386 TI - Tachycardia-induced changes in Na+ current in a chronic dog model of atrial fibrillation. AB - We have previously shown that chronic rapid atrial activation (400 bpm) reduces atrial conduction velocity in dogs, contributing to the development of a substrate supporting sustained atrial fibrillation (AF). However, the cellular and ionic mechanisms underlying these functional changes have not been defined. We applied whole-cell patch-clamp techniques to atrial myocytes from dogs subjected to atrial pacing at 400 bpm for 7 days (P7, n = 6) and 42 days (P42, n = 5) and compared the results with those from sham-operated dogs similarly instrumented but without pacemaker activation (P0, n = 6). Rapid atrial pacing allowed for the induction of sustained AF in 67% and 100% of dogs paced for 7 and 42 days, respectively, and significantly decreased conduction velocity under P7 and P42 conditions. In dogs paced for 7 days, Na+ current (INa) density was reduced by 28% at -40 mV (P < .0001, n = 59 cells). INa changes were even more decreased under P42 conditions, by approximately 52% at -40 mV (P < .0001): from 78.7 +/- 4.6 pA/pF (P0, n = 28 cells) to -37.7 +/- 3.0 pA/pF (P42, n = 43 cells). INa was significantly reduced at all voltages ranging from -65 to -10 mV. Voltage dependent activation and inactivation properties, activation kinetics, and recovery from inactivation were not altered by rapid atrial pacing; however, inactivation kinetics were slowed. AF duration was related to mean INa in each dog (r2 = .573, P < .001). We conclude that rapid atrial activation significantly reduces both conduction velocity and INa density. Since INa is a major determinant of conduction velocity, our data point to INa reduction as a potentially important mechanism contributing to the substrate for AF in this model. PMID- 9400387 TI - Molecular determinants of stereoselective bupivacaine block of hKv1.5 channels. AB - Enantiomers of local anesthetics are useful probes of ion channel structure that can reveal three-dimensional relations for drug binding in the channel pore and may have important clinical consequences. Bupivacaine block of open hKv1.5 channels is stereoselective, with the R(+)-enantiomer being 7-fold more potent than the S(-)-enantiomer (Kd = 4.1 mumol/L versus 27.3 mumol/L). Using whole-cell voltage clamp of hKv1.5 channels and site-directed mutants stably expressed in Ltk- cells, we have identified a set of amino acids that determine the stereoselectivity of bupivacaine block. Replacement of threonine 505 by hydrophobic amino acids (isoleucine, valine, or alanine) abolished stereoselective block, whereas a serine substitution preserved it [Kd = 60 mumol/L and 7.4 mumol/L for S(-)- and R(+)-bupivacaine, respectively]. A similar substitution at the internal tetraethylammonium binding site (T477S) reduced the affinity for both enantiomers similarly, thus preserving the stereoselectivity [Kd = 45.5 mumol/L and 7.8 mumol/L for S(-)- and R(+)-bupivacaine, respectively]. Replacement of L508 or V512 by a methionine (L508M and V512M) abolished stereoselective block, whereas substitution of V512 by an alanine (V512A) preserved it. Block of Kv2.1 channels, which carry valine, leucine, and isoleucine residues at T505, L508, and V512 equivalent sites, respectively, was not stereoselective [Kd = 8.3 mumol/L and 13 mumol/L for S(-)- and R(+) bupivacaine, respectively]. These results suggest that (1) the bupivacaine binding site is located in the inner mouth of the pore, (2) stereoselective block displays subfamily selectivity, and (3) a polar interaction with T505 combined with hydrophobic interactions with L508 and V512 are required for stereoselective block. PMID- 9400388 TI - Reduced adenosine A1 receptor and G alpha protein coupling in rat ventricular myocardium during aging. AB - Adenosine A1 receptor (A1-AdoR) function in rat ventricles has previously been shown to decrease with age. In the present study, using the ligand [3H]8 cyclopentyl-1,3-dipropylxanthine ([3H]DPCPX) and coimmunoprecipitation of A1 AdoRs with their associated G proteins, we determined the specific binding of A1 AdoR and A1-AdoR/G protein coupling in ventricular myocardium of 6- to 24-month old Fischer 344 rats. The densities (Bmax) of A1-AdoRs were 5.8 +/- 0.8 fmol/mg protein in 6-month-old rats and 6.1 +/- 1.4 fmol/mg protein in 24-month-old rats, and the dissociation constants (Kd) were 0.32 +/- 0.04 nmol/L in 6-month-old rats and 0.34 +/- 0.05 nmol/L in 24-month-old rats (P > .05). Analysis of the dose dependent displacement of [3H]DPCPX binding by the selective A1-receptor agonist, N6-p-sulfophenyladenosine (SPA), yielded two affinity binding sites in both 6- and 24-month-old rats. However, the proportion of high-affinity A1-AdoRs was significantly lower in 24-month-old rats (23.5%) compared with 6-month-old rats (54.9%) (P < .05). In solubilized ventricular membranes, specific [3H]DPCPX binding sites were detected in immunoprecipitates of G alpha i3 and G alpha o antisera but not with antibodies for other G alpha proteins. The basal coimmunoprecipitation of A1-AdoR with G alpha i3 and G alpha o proteins decreased by 22% and 21%, respectively, in ventricular membranes of 24-month-old rats compared with that in 6-month-old animals. A1-AdoR stimulation with SPA increased the coprecipitation of A1-AdoR with G alpha i3 and G alpha o proteins by 287% and 245%, respectively, in 6-month-old rats but only by 129% and 140%, respectively, in 24-month-old rats (P < .01). In the absence of changes in A1-AdoR density and G alpha protein levels, an age-related decline in high-affinity A1-AdoR binding sites and a reduction in the association of A1-AdoR with G alpha proteins suggest that the age-related decrease in ventricular A1-AdoR-mediated response is related to a reduction in the coupling between A1-AdoR and their G proteins. PMID- 9400389 TI - Cardioprotective effect of diazoxide and its interaction with mitochondrial ATP sensitive K+ channels. Possible mechanism of cardioprotection. AB - Previous studies showed a poor correlation between sarcolemmal K+ currents and cardioprotection for ATP-sensitive K+ channel (KATP) openers. Diazoxide is a weak cardiac sarcolemmal KATP opener, but it is a potent opener of mitochondrial KATP, making it a useful tool for determining the importance of this mitochondrial site. In reconstituted bovine heart KATP, diazoxide opened mitochondrial KATP with a K1/2 of 0.8 mumol/L while being 1000-fold less potent at opening sarcolemmal KATP. To compare cardioprotective potency, diazoxide or cromakalim was given to isolated rat hearts subjected to 25 minutes of global ischemia and 30 minutes of reperfusion. Diazoxide and cromakalim increased the time to onset of contracture with a similar potency (EC25, 11.0 and 8.8 mumol/L, respectively) and improved postischemic functional recovery in a glibenclamide (glyburide) reversible manner. In addition, sodium 5-hydroxydecanoic acid completely abolished the protective effect of diazoxide. While-myocyte studies showed that diazoxide was significantly less potent than cromakalim in increasing sarcolemmal K+ currents. Diazoxide shortened ischemic action potential duration significantly less than cromakalim at equicardioprotective concentrations. We also determined the effects of cromakalim and diazoxide on reconstituted rat mitochondrial cardiac KATP activity. Cromakalim and diazoxide were both potent activators of K+ flux in this preparation (K1/2 values, 1.1 +/- 0.1 and 0.49 +/- 0.05 mumol/L, respectively). Both glibenclamide and sodium 5-hydroxydecanoic acid inhibited K+ flux through the diazoxide-opened mitochondrial KATP. The profile of activity of diazoxide (and perhaps KATP openers in general) suggests that they protect ischemic hearts in a manner that is consistent with an interaction with mitochondrial KATP. PMID- 9400390 TI - Enhanced Na(+)-Ca2+ exchange in the infarcted heart. Implications for excitation contraction coupling. AB - Cellular Ca2+ regulation is abnormal in diseased hearts. We designed this study to assess the role of the Na(+)-Ca2+ exchanger in excitation-contraction coupling in surviving myocardium of the infarcted heart. We measured cellular contractions and whole-cell currents in single left ventricular myocytes isolated from the hearts of rabbits with healed myocardial infarction (MI). Eight weeks after MI, rabbits had left ventricular dysfunction without overt heart failure. Myocytes isolated from regions adjacent to the infarcted zone were significantly longer than cells from control hearts. At low stimulation rates (0.5 Hz), the amplitude of field-stimulated contractions was increased (11.6 +/- 0.5% versus 10.2 +/- 0.6% resting cell length), whereas the time to peak shortening and action potential duration were prolonged in the MI cells. When stimulation frequency was increased to 2.0 Hz, cellular shortening did not change or decreased in myocytes from infarcted hearts, whereas control cells had a positive shortening-interval relationship. Cells from infarcted hearts had a significantly decreased (31%) L type Ca2+ current (ICa) density but no change in the current-voltage relationship or the kinetics of ICa inactivation. Maximal Na(+)-Ca2+ exchange current density was significantly increased (32%) in the cells from infarcted hearts. Sarcoplasmic reticulum (SR) Ca2+ content during a stable train of contractions, as estimated from caffeine-induced inward currents, was slightly increased (P = NS) in the MI myocytes. To determine whether Na(+)-Ca2+ exchange influenced SR Ca2+ content, cells were clamped at potentials between -70 and +90 mV for 400 ms. The amplitude of the contraction during a subsequent clamp step to +10 mV was then measured as an index of SR loading that occurred during the preceding clamp step. Steps to positive potentials produced greater augmentation of the subsequent contraction in MI than in control myocytes. In myocytes from the infarcted heart, increased activity of the Na(+)-Ca2+ exchanger may promote Ca2+ entry or decrease Ca2+ extrusion. This relative augmentation of inward Ca2+ flux by the exchanger may enhance SR Ca2+ loading and thus support contractility that would otherwise be impaired as a result of decreased Ca2+ current. However, Ca2+ influx by the exchanger may contribute to the prolongation of contractions in myocytes from infarcted hearts. PMID- 9400392 TI - Students get gritty introduction to reality of HIV/AIDS. PMID- 9400391 TI - The protective effect of late preconditioning against myocardial stunning in conscious rabbits is mediated by nitric oxide synthase. Evidence that nitric oxide acts both as a trigger and as a mediator of the late phase of ischemic preconditioning. AB - Seventy-four conscious rabbits undergoing a sequence of six 4-minute coronary occlusion/4-minute reperfusion cycles for 3 consecutive days (days 1, 2, and 3) were assigned to nine groups. In group I (controls, n = 8), the recovery of systolic wall thickening (WTh) after the sixth reperfusion was markedly improved on days 2 and 3 compared with day 1, indicating late preconditioning (PC) against myocardial stunning; the total deficit of WTh after the sixth reperfusion was reduced by 56% on day 2 and 50% on day 3 compared with day 1 (P < .01). Administration on day 2 of the nonselective NO synthase (NOS) inhibitor N omega nitro-L-arginine (L-NA) (group II, n = 8) or of the selective inducible NOS inhibitors aminoguanidine (AG) (group IV, n = 8) and S-methylisothiourea sulfate (SMT) (group VI, n = 6) completely abrogated late PC against stunning on day 2. On day 3, the expected PC effect became manifest in all groups. Administration of L-NA, AG, or SMT on day 1 (groups III [n = 7], V [n = 6], and VII [n = 5], respectively) had no discernible effect on the deficit of WTh on day 1, indicating that these agents do not augment the severity of myocardial stunning in nonpreconditioned myocardium. In group VIII (n = 7), the abrogation of late PC by SMT on day 2 was completely reversed by the concomitant administration of L arginine (595 mg/kg IV), indicating that it was not due to nonspecific NOS unrelated actions. Administration of L-arginine alone on day 2 (group IX [n = 5]) had no effect on the deficit of WTh. Furthermore, administration of L-NA on day 1 (group III) prevented the appearance of the PC effect on day 2, whereas AG (group V) and SMT (group VI) did not, suggesting that the development of late PC on day 1 is triggered by the endothelial (type III) isoform of NOS. This study demonstrates that three structurally different NOS inhibitors (L-NA, AG, and SMT), given 24 hours after the PC ischemia, consistently abrogate late PC against myocardial stunning in conscious rabbits, indicating that this cardioprotective effect is mediated by the activity of NOS. The results obtained with AG and SMT specifically implicate the inducible (type II) isoform as the mediator of the protection on day 2. Previous studies have shown that NO triggers the development of late PC. The present results indicate that NO plays a dual role in late PC against stunning, acting initially as the trigger and subsequently as the mediator of the protection. PMID- 9400393 TI - Ontario's HSOs have not failed! PMID- 9400394 TI - Ontario's HSOs have not failed! PMID- 9400395 TI - CPGs: to reach the unreachable goal? PMID- 9400396 TI - Addressing needle-stick concerns. PMID- 9400397 TI - The US attack on Cuba's health. PMID- 9400398 TI - The US attack on Cuba's health. PMID- 9400399 TI - The US attack on Cuba's health. PMID- 9400400 TI - Shedding light on sunscreen use. PMID- 9400401 TI - Signing up with ADD. PMID- 9400402 TI - Cervical cancer screening. PMID- 9400404 TI - Why? PMID- 9400403 TI - The many faces of pheochromocytoma. PMID- 9400405 TI - Why? PMID- 9400406 TI - Induction of labour versus expectant management for prelabour rupture of the membranes at term: an economic evaluation. TERMPROM Study Group. Term Prelabour Rupture of the Membranes. AB - BACKGROUND: As the interval between rupture of the fetal membranes at term and delivery increases, so may the risk of fetal and maternal infection. Recently the TERMPROM (Term Prelabor Rupture of the Membranes) Study Group reported the results of a randomized controlled trial comparing 4 management strategies: induction with oxytocin (IwO), induction with prostaglandin (IwP), and expectant management and induction with either oxytocin (EM-O) or prostaglandin (EM-P) if complications developed. The study found no statistically significant differences in neonatal infection and cesarean section rates between any of the 4 groups. OBJECTIVE: To conduct an economic evaluation comparing the cost of (a) IwO and EM O, (b) IwP and EM-P and (c) IwO and IwP. DESIGN: An economic analysis, conducted alongside the clinical trial, using a third-party payer perspective. Analysis included all treatment costs incurred for both the mother and the baby. Information on health care utilization and outcomes was collected for all study participants. Three countries (Canada, the United Kingdom and Australia), corresponding to the largest study recruitment, were chosen for calculation of unit costs. For each country, the base, low and high estimates of unit cost for each service item were generated. Intention-to-treat analysis. Extensive statistical and sensitivity analyses were performed. RESULTS: The median cost of IwO per patient was significantly lower statistically than that of EM-O and IwP. This result held in all 3 countries compared -$114 and -$46 in Canada, -113 Pounds and -63 Pounds in the UK, and -A$30 and -A$49 in Australia) and after an extensive sensitivity analysis. There was no statistically significant difference in median cost per patient between IwP and EM-P. CONCLUSION: Although the clinical results of the TERMPROM study did not find IwO to be preferable to the other treatment alternatives, the economic evaluation found it to be less costly. However, these cost differences, even though statistically significant, are not likely to be important in many countries. When this is the case, the authors recommend that women be offered a choice between management strategies. PMID- 9400407 TI - Alcohol disorders in Canada as indicated by the CAGE questionnaire. AB - OBJECTIVE: To describe alcohol disorders in the general Canadian population, using as a standard indicator the CAGE questionnaire (Have you felt you needed to cut down on your drinking? Have you felt annoyed by criticism of your drinking? Have you felt guilty about drinking? Have you felt you needed a drink first thing in the morning [eye-opener]?). DESIGN: Secondary analysis of data from Canada's Alcohol and Other Drugs Survey (CADS), a national telephone survey conducted in 1994 of a representative sample of 12,155 people aged 15 years or more. PARTICIPANTS: The CAGE questionnaire was administered to 5894 drinkers who had consumed alcohol in the 12 months before the CADS survey. MAIN OUTCOME MEASURES: Respondents with positive (2 or more affirmative responses) and negative results on the CAGE questionnaire were compared as to demographic characteristics, alcohol consumption and harmful consequences of their drinking. Independent predictors of a positive result were identified by means of logistic regression analysis. RESULTS: A total of 5.8% of CAGE-tested current drinkers had a positive result on the past-year CAGE in 1994. The proportion of respondents reporting alcohol-related problems in one or more areas of their life was 7 times greater among drinkers with a positive result on the CAGE questionnaire than among those with a negative result (66.8% v. 9.5%) (p < 0.0001). When all demographic characteristics were controlled for simultaneously, male sex, residence in the Atlantic provinces, Quebec or the Prairies, single/never married or divorced/separated marital status, and low education level were found to be independent risk factors for a positive result on the CAGE questionnaire. About 85% of the respondents with a positive result had not sought help for their drinking. Applying the estimated sensitivity and specificity of the CAGE questionnaire in detecting alcohol dependence, as per criteria of the Diagnostic and Statistical Manual, in a general US population, the authors estimated that 4.1% of Canadians had an alcohol dependence in 1994. CONCLUSION: The large proportion of current drinkers with a positive result on the CAGE questionnaire who did not seek help for their drinking underscores the need for identification and brief interventions by physicians. Further research is needed to elucidate the underlying reasons for regional differences in CAGE status. PMID- 9400408 TI - Responding to our abused patients. PMID- 9400409 TI - Premature rupture of membranes at term: a medical and economic rationale for active management. PMID- 9400410 TI - Alcohol disorders in Canada: the need for intervention. PMID- 9400411 TI - Avoiding the mismeasurement of medicine and improving care. PMID- 9400412 TI - More than meets the eye: recognizing and responding to spousal abuse. PMID- 9400413 TI - Medical classification systems in Canada: moving toward the year 2000. AB - The use of different standards for coding diagnoses and procedures has been identified as a major obstacle to the collection and analysis of data across the various jurisdictions in Canada. In this article the authors briefly describe the current and future situation of medical classification systems in Canada and discuss some of the potential benefits and implications of adopting the 10th revision of the International Statistical Classification of Diseases and Related Health Problems and a revised procedure classification, the Canadian Classification of Health Interventions, as national standards for classification systems in Canada. They further describe some of the key features of the proposed new classification systems and highlight some of the actions being taken by the Canadian Institute for Health Information to support implementation of these standards in Canada over the next few years. PMID- 9400414 TI - A curriculum for the times: an experiment in teaching health policy to residents in family medicine. AB - The Department of Family Medicine at Queen's University in Kingston, Ont., recently undertook a pilot project to familiarize residents in family medicine with physician-related health policy issues. The objective of the project was to ease the residents' transition into practice and to equip them to participate effectively in future policy debates. All first-year residents assigned to a 4 month clinical rotation in the Department of Family Medicine took part in the program, which consisted of 5 weekly 1-hour lecture and discussion sessions. The program was offered as one component of the 130-hour core curriculum for first year residents. Participants evaluated the program as highly informative and extremely relevant to their career plans. The authors conclude that health policy is a subject that can be incorporated into the core curriculum of residency training programs. PMID- 9400415 TI - Amantadine use in influenza outbreaks in long-term care facilities. PMID- 9400416 TI - MDs have key role in bringing ugly secret of wife abuse out of closet. PMID- 9400417 TI - Mother's rights can't be infringed to protect fetus, Supreme Court's landmark ruling states. AB - Cases involving child abuse have received wide coverage lately, as has a case involving possible risk to a fetus because of a mother's addiction to solvents. Lawyer Karen Capen discusses the legal issues facing doctors over the reporting of child abuse and outlines their obligations and responsibilities. PMID- 9400418 TI - Move into hospital sector another sign of complementary medicine's growing popularity. AB - Growing demand has led some Canadian hospitals to offer alternative therapies to patients, even though many physicians still question their efficacy. Anita Elash visited Toronto's Sunnybrook Health Science Centre, where staff physicians have been debating the issue. One physician said hospitals have no choice but to offer the treatments. "If you believe in the primacy of patients making their own decisions and you believe in the fundamental of informed consent, you cannot deny them access to this treatment." PMID- 9400419 TI - Work by Alberta researchers may free MDs from awkward fitness-to-drive decisions. AB - An Alberta researcher has developed tests that determine whether a senior citizen with cognitive impairment is fit to drive. The issue is important to the medical profession because these drivers sometimes cause accidents and physicians often have to decide whether they should still be allowed to get behind the wheel. PMID- 9400420 TI - Common cancers--immunotherapy and multidisciplinary therapy: Parts III and IV. AB - The refractoriness of many solid tumors to cytotoxic chemotherapy has led to the exploration of new therapeutic modalities, including immunotherapy. Immunotherapy does not have a direct cytotoxic effect on the cancer cell but is an attempt to promote rejection of the tumor by the host, chiefly through the cellular arm of the immune system. The clinical success with immunotherapy (primarily adoptive immunotherapy) among patients with unresectable malignant melanoma and cancer of the kidney has not been marked by the large numbers of patients responding but by occasional dramatic effectiveness of therapy for these cancers, which usually are refractory to chemotherapy. Long-lasting responses and even complete disappearance of all known metastases are possible for a small percentage of patients with melanoma or renal cell carcinoma who undergo immunotherapy. A reasonable approach for patients with good performance status (no symptoms or ambulatory with symptoms but not bedridden) is entrance to clinical trials, especially trials examining adoptive or active immunotherapy for melanoma or adoptive immunotherapy for renal cancer. The overall treatment of patients with cancer has changed. Primary-care physicians detect almost all cancers. The days when "taking it out" is the best we could offer a patient are over. As we learn more about the use of adjuvant or neoadjuvant chemotherapy and radiation therapy, it is likely one or both of these modalities will be incorporated into the treatment of additional solid tumors previously managed solely with surgical resection. Increasingly, additional therapy is being given for earlier-stage disease as we define how to maximize the potential for cure with minimal toxicity. Many new therapies are on the horizon, including the use of noncytotoxic treatments as an adjunct to a surgical procedure. Such therapies include the use of angiogenesis inhibitors, tumor vaccines, and immunotherapy. Now and in the future, patients will be best served when treated in an environment that can integrate medical, surgical, and radiation oncology actively. PMID- 9400421 TI - Carboxy-terminal domain mediates assembly of the voltage-gated rat ether-a-go-go potassium channel. AB - The specific assembly of subunits to oligomers is an important prerequisite for producing functional potassium channels. We have studied the assembly of voltage gated rat ether-a-go-go (r-eag) potassium channels with two complementary assays. In protein overlay binding experiments it was shown that a 41-amino-acid domain, close to the r-eag subunit carboxy-terminus, is important for r-eag subunit interaction. In an in vitro expression system it was demonstrated that r-eag subunits lacking this assembly domain cannot form functional potassium channels. Also, a approximately 10-fold molar excess of the r-eag carboxy-terminus inhibited in co-expression experiments the formation of functional r-eag channels. When the r-eag carboxy-terminal assembly domain had been mutated, the dominant-negative effect of the r-eag carboxy-terminus on r-eag channel expression was abolished. The results demonstrate that a carboxy-terminal assembly domain is essential for functional r-eag potassium channel expression, in contrast to the one of Shaker-related potassium channels, which is directed by an amino-terminal assembly domain. PMID- 9400422 TI - Acute and long-lasting effects of neonatal hypoxia on (+)-3-[125I]MK-801 binding to NMDA brain receptors. AB - The NMDA receptor subtype is the major excitatory mediator for glutamate neurotoxicity. To assess its participation in the noxious effects of postnatal hypoxia, we have characterized the binding of the ionophoric marker of NMDA receptor, dizocilpine (MK-801). Binding of (+)-3-[125I]MK-801 to NMDA brain receptors under nonequilibrium conditions was quantified by in vitro autoradiography in rats exposed to hypoxia induced by 93% N2/6.5% O2 exposure for 70 min on Postnatal Day 4. Acute and long-lasting effects were investigated at 4 h after injury and on Postnatal Day 40. At the acute stage, a transient decrease in binding was found in several specific brain areas, hypothalamus, amygdaloid nuclei, entorhinal cortex, perirhinal cortex, and hippocampus, and no differences were found in temporal cortex, thalamus, and geniculate nucleus, when compared to sham-treated animals. At this early age, there was no increase of binding when slices from both groups were incubated in the presence of glutamate and glycine (Glu/Gly), positive allosteric modulators of MK-801 binding. In the 40-day-old brains, the binding to the NMDA receptors of hypoxiatreated animals was not different with respect to controls in most of the areas studied, but the Glu/Gly stimulation of binding in hypoxic rats showed a reduced, or absent, response to the allosteric modulators. In contrast, control rats showed a remarkable increase of the specific binding induced by the presence of the modulators in the incubation buffer. Binding of (+)-3-[125I]MK-801 was also performed at a higher concentration to clarify whether the altered response to Glu/Gly may be due to differences in the number of channels; however, the density of NMDA receptors at this concentration was similar in both control and hypoxia-treated rats. We conclude that the effect of exposure of newborn rats to hypoxia can generate acute and long-lasting effects on the NMDA receptor. The deleterious action of this kind of noxa on the CNS could be exerted by interference with normal glutamatergic transmission and hence over normal growth and development. PMID- 9400423 TI - Spinal cord transection--no loss of distal ventral horn neurons. Modern stereological techniques reveal no transneuronal changes in the ventral horns of the mouse lumbar spinal cord after thoracic cord transection. AB - Anterograde transneuronal degeneration is caused by the loss of afferent input to the nerve cells and may occur in a number of neuronal systems. Transection of the adult spinal cord, causing anterograde transneuronal degeneration in ventral horn neurons, distal to the lesion, has been reported by some authors, while others contend that no such changes take place. The present study was undertaken in order to investigate whether transection of adult mouse thoracic spinal cord induces neuronal death in the ventral horns distal to the lesion. By means of modern stereological techniques such as the optical dissector, the total number of cells in the lumbar ventral horns was estimated 7 weeks after transection. The mean numbers of neurons and glial and endothelial cells were 82,000 versus 89,000, 259,000 versus 301,000, and 129,000 versus 144,000 in the transected (n = 6) and sham-operated animals (n = 5), respectively. These differences were not statistically significant. Furthermore, neuronal soma volume was estimated by another stereological method, the vertical rotator. Mean neuronal soma volume was not significantly different between transected (2762 microns 3) and sham-operated (2617 microns 3) mice. Although no reduction in cell number or neuronal soma volume was observed, the mean volume of the ventral horns in the lumbar segments was significantly less in transected than in sham-operated animals, 2.49 mm3 versus 3.05 mm3 (P < 0.05). In conclusion, the transection of adult mouse thoracic spinal cord does not induce neuronal degeneration in the lumbar ventral horns. PMID- 9400424 TI - Re: Unstable trinucleotide repeats in Alzheimer's disease? PMID- 9400425 TI - Autologous cell transplantation for urologic reconstruction. PMID- 9400426 TI - Interpretation of free prostate specific antigen clinical research studies for the detection of prostate cancer. AB - PURPOSE: We reviewed the use of percent free prostate specific antigen (PSA) to enhance specificity of PSA testing and aid in the discrimination of benign and malignant prostate disease. We present proposed percent free PSA cut points and probability factors, and discuss factors that are believed to affect study outcomes and conclusions. MATERIALS AND METHODS: We reviewed the literature with respect to PSA and free PSA with particular emphasis on clinical use of percent free PSA and factors that may affect study outcomes. RESULTS: Percent free PSA may increase the specificity of PSA testing without sacrificing the cancer detection rate. Differences in study designs and subject populations may account for the confusion in the current literature. Specific factors that may influence study outcomes include sample size, PSA range, age, race, digital rectal examination findings, prostate size, tumor size and pathology, as well as treatment history, sample collection and storage conditions, and the particular assays used to determine free and total PSA values. CONCLUSIONS: The use of percent free PSA to enhance the specificity of prostate cancer screening is thought to provide useful information to aid in the differentiation of benign and malignant prostate diseases. There is evidence to suggest a benefit cost advantage to a tailored biopsy approach based on percent free PSA. However, statistically valid multisite clinical trials that take into account influencing factors are needed to set assay specific cut points and probability determinations. PMID- 9400428 TI - Holmium:YAG lithotripsy yields smaller fragments than lithoclast, pulsed dye laser or electrohydraulic lithotripsy. AB - PURPOSE: The mechanism of lithotripsy differs among electrohydraulic lithotripsy, mechanical lithotripsy, pulsed dye lasers and holmium:YAG lithotripsy. It is postulated that fragment size from each of these lithotrites might also differ. This study tests the hypothesis that holmium:YAG lithotripsy yields the smallest fragments among these lithotrites. MATERIALS AND METHODS: We tested 3F electrohydraulic lithotripsy, 2 mm. mechanical lithotripsy, 320 microns pulsed dye lasers and 365 microns. holmium:YAG fiber on stones composed of calcium hydrogen phosphate dihydrate, calcium oxalate monohydrate, cystine, magnesium ammonium phosphate and uric acid. Fragments were dessicated and sorted by size. Fragment size distribution was compared among lithotrites for each composition. RESULTS: Holmium:YAG fragments were significantly smaller on average than fragments from the other lithotrites for all compositions. There were no holmium:YAG fragments greater than 4 mm., whereas there were for the other lithotrites. Holmium:YAG had significantly greater weight of fragments less than 1 mm. compared to the other lithotrites. CONCLUSIONS: Holmium:YAG yields smaller fragments compared to electrohydraulic lithotripsy, mechanical lithotripsy or pulsed dye lasers. These findings imply that fragments from holmium:YAG lithotripsy are more likely to pass without problem compared to the other lithotrites. Furthermore, the significant difference in fragment size adds evidence that holmium:YAG lithotripsy involves vaporization. PMID- 9400427 TI - Historical milestones regarding torsion of the scrotal organs. AB - PURPOSE: The clinical syndrome of the acute scrotum, whereby the spermatic cord or appendix testis becomes twisted, commonly affects young men. Our knowledge of this condition, however, is of relatively recent origin. MATERIALS AND METHODS: We performed an historical survey of torsion of the scrotal organs dating back to 1703. In Bologna in 1703 Morgagni observed the first hydatid on the caput epididymis. He described 10 hydatid cases of the testis and epididymis producing, in his opinion, the fluid of hydroceles. The first illustration of appendix testis dates from Cooper 1841. Later these testicular appendages, or hydatids, were shown to be vestigial remnants of either the mullerian duct or the wolffian structures, depending on location. Actual torsion of the appendix testis was mentioned by Ombredanne in 1913 but the first case report was published in 1922 by Colt. Appendix testis torsion was first schematically illustrated in 1923 by Mouchet and was characterized by Dix in 1931 in a manner that is still valid today. Interestingly, the great majority of case reports since 1932 have originated from America. In 1810 Hunter described a typical case of testicular torsion, and in 1840 Delasiauve presented the first case of surgically treated testicular torsion. A schematic and original illustration of a contorted undescended testis was published in 1894 by Lauenstein. RESULTS: Testicular torsion was, and still is, a true urological emergency but the historical survival rate of the testis was extremely low. Searching for improvement in clinical diagnosis, physicians have noted helpful specific signs, 1 of which is that the period of ischemia determines loss of the testis. CONCLUSIONS: The historical development of diagnosis of torsion of the appendix testis and spermatic cord highlights the ever present need for careful examination, a high index of suspicion and timely therapy. PMID- 9400429 TI - Efficacy and cost-effectiveness of extracorporeal shock wave lithotripsy for solitary lower pole renal calculi. AB - PURPOSE: We determined the efficacy of extracorporeal shock wave lithotripsy monotherapy and compared its cost-effectiveness with percutaneous nephrolithotomy for the management of lower pole renal calculi. MATERIALS AND METHODS: The efficacy (stone-free rates at 3-months posttreatment) of shock wave lithotripsy with the modified Dornier HM3* machine was determined retrospectively in 114 patients with solitary lower pole renal calculi. Using cost data available from patient billing charges and efficacy data from the literature, the cost effectiveness for percutaneous nephrolithotomy and shock wave lithotripsy as primary therapy was evaluated. To make this cost-effectiveness comparison, we developed a decision analysis model in which a patient in whom primary therapy failed was rendered stone-free with a secondary percutaneous nephrolithotomy procedure. RESULTS: The stone-free rates of solitary lower pole stones with a size range of less than 10, 11 to 20 and greater than 20 mm. were 76, 74 and 33%, respectively, with a single shock wave lithotripsy treatment. Based on average treatment costs for shock wave lithotripsy and percutaneous nephrolithotomy, the model results show that for stone sizes less than 2 cm. primary lithotripsy therapy followed by nephrolithotomy for failed cases is the least costly approach. For stone sizes greater than 2 cm. primary percutaneous nephrolithotomy may be more cost-effective. CONCLUSIONS: Whereas shock wave lithotripsy with the Dornier HM3 should be considered the initial treatment choice for most lower pole stones less than 2 cm., primary percutaneous nephrolithotomy should be considered for stones larger than 2 cm. PMID- 9400430 TI - A placebo controlled study of mycophenolate mofetil used in combination with cyclosporine and corticosteroids for the prevention of acute rejection in renal allograft recipients: 1-year results. The European Mycophenolate Mofetil Cooperative Study Group. AB - PURPOSE: We performed a randomized, double-blind, multicenter, placebo controlled study to compare the efficacy and safety of 2 oral doses of mycophenolate mofetil with placebo for prevention of acute rejection episodes following first or second cadaveric renal allograft transplantation. MATERIALS AND METHODS: A total of 491 patients were enrolled in the study and randomly allocated to receive placebo (166), 1 gm. mycophenolate mofetil twice daily (165) or 1.5 gm. mycophenolate mofetil twice daily (160). Patients were given concomitant immunosuppression with cyclosporine and corticosteroids. Treatment with mycophenolate mofetil was initiated within 72 hours of transplantation and was continued for at least 1 year. RESULTS: The percentages of patients who experienced biopsy proved rejection or withdrew early from the trial for any reason were significantly reduced with 2 gm. (30.3%) and 3 gm. (38.8%) mycophenolate mofetil compared to placebo (56.0%) (p < 0.001). The biopsy proved rejection rates of the placebo, and 2 gm. and 3 gm. mycophenolate mofetil treatment arms were 46.4, 17.6 and 13.8%, respectively. There were fewer patients in the 2 gm. (28.5%) and 3 gm. (24.4%) mycophenolate mofetil groups compared to the placebo (51.8%) group, who received full courses of corticosteroids or antilymphocyte agents for treatment of rejection episodes in the first 6 months after renal transplantation. There was a greater incidence of gastrointestinal adverse events, leukopenia and opportunistic events in the mycophenolate mofetil treatment groups. CONCLUSIONS: Mycophenolate mofetil was shown to reduce significantly the number of patients who experienced biopsy proved rejection episodes or treatment failure during the first year after renal transplantation, and was well tolerated. PMID- 9400431 TI - Endoluminal stent placement after percutaneous transluminal angioplasty in the treatment of post-transplant renal artery stenosis. AB - PURPOSE: We report our experience with endoluminal stent placement after percutaneous transluminal angioplasty for the treatment of post-transplant renal artery stenosis. MATERIALS AND METHODS: From October 1992 to September 1996, 8 stents were successfully implanted in 7 patients affected by resistant transplant renal artery stenosis. All transplanted kidneys were procured from cadaver donors. The patients were routinely evaluated with duplex sonography and the median interval between transplantation and stenosis detection was 7.4 months (range 0.5 to 17). When serious renal stenosis was diagnosed (greater than 50%), selected angiography and percutaneous transluminal angioplasty were performed. In 8 cases (7 patients) an endoluminal metallic Palmaz stent was placed in the site of the restenosis. One patient received 2 stents repeatedly positioned in different stenosis sites. RESULTS: No major complications occurred. Clinical outcome was positive in 5 patients (71.4%) and Stenosis recurred in 2 (28.5%) (less than 50% and less than 35%, respectively). Median followup after stent placement was 14.8 months (range 1 to 37). CONCLUSIONS: Percutaneous endoluminal stent procedures after resistant transplant renal artery stenosis or for ex novo treatment for severe anastomotic stenoses appears to be promising. Longer followup periods are necessary for true evaluation of this procedure. PMID- 9400433 TI - Bilateral open transperitoneal cyst reduction surgery for autosomal dominant polycystic kidney disease. AB - PURPOSE: We reviewed our experience with open transperitoneal bilateral renal cyst reduction surgery in patients with symptomatic autosomal dominant polycystic kidney disease to define perioperative morbidity and mortality, and to suggest that others consider this mode of therapy when more conservative methods fail to provide relief from pain or early satiety. MATERIALS AND METHODS: A total of 28 patients underwent 30 transperitoneal bilateral renal cyst reduction decompression operations between May 1987 and June 1996. Ten procedures included surgical treatment of concomitant liver cysts (8 by liver cyst marsupialization and 2 by partial hepatic resection). Records were reviewed for hospital stay, perioperative morbidity, changes in renal function and hypertension control. RESULTS: Hospitalization averaged 9 days. Treatment of hepatic cysts, age and renal insufficiency did not extend hospitalization. A transient reduction in renal function occurred after 20 procedures. The most frequently encountered perioperative morbid events were ileus in 4 patients and cardiac arrhythmias in 3. The most significant complications were myocardial infarction in 1 patient and fatal adult respiratory distress syndrome after partial liver resection in another. Preoperative renal insufficiency, age and treatment of hepatic cysts were not associated with increased morbidity. Six patients had improvement in hypertension and none had sepsis. CONCLUSIONS: Bilateral transperitoneal renal cyst reduction surgery is a relatively safe and effective treatment for individuals with symptomatic polycystic kidney disease in whom more conservative therapies have failed. PMID- 9400432 TI - Urological complications after kidney-pancreas transplantation. AB - PURPOSE: Urological complications are common after kidney-pancreas transplantation. Predictors of urological complication after transplantation have not been established. We studied the impact of urological complications on allograft function. In addition we evaluated age at transplantation, diabetic years before transplantation and preoperative bladder function as predictors of allograft pancreatitis, postoperative retention and urine leaks. MATERIALS AND METHODS: Urological complications in 65 cases (38 men, 27 women, mean diabetic years 21 +/- 6, mean age 33 +/- 7 years) who had transplants between December 1987 and January 1995 were reviewed. Preoperative urodynamics in 50 patients (77%) and voiding cystourethrogram in 40 (62%) were analyzed. Kidney-pancreas transplantation was completed using bladder drainage techniques. RESULTS: Mean followup was 44 +/- 27 months (median 40, range 1 to 93). Urological complications in 51 patients (79%) included urinary tract infection in 59%, hematuria in 26%, allograft pancreatitis in 19%, duodenal leaks in 17%, ureteral lesions in 9% and urethral lesions in 6%. Eleven duodenal leaks (8 leaks in less than 1 month) required surgical treatment. Nine leaks recurred in 7 patients. Allograft pancreatitis occurred 32 times (range 1 to 9) in 12 patients. Three patients had ureteral obstruction and 3 had ureteral leaks. Preoperative urodynamics included detrusor hyperreflexia in 8 patients, detrusor areflexia in 19, indeterminate in 5 and normal in 18. The 1-year patient, kidney and pancreatic allograft survival rates were 92, 91 and 86%; 2-year survival rates were 89, 88 and 80%, and 5-year survival rates were 61, 59 and 55%, respectively. CONCLUSIONS: Urological complications were common after transplantation but did not adversely affect allograft survival in our series. Age at transplantation, diabetic years preceding transplantation and preoperative bladder function were not significantly correlated with allograft pancreatitis, postoperative urinary retention or urine leaks. A prospective analysis of postoperative bladder function should be completed to improve understanding and possibly reduce morbidity of urological complications after transplantation. PMID- 9400434 TI - Monoclonal antibodies against renal tumors: the potential application in discrimination of ambiguous adenocarcinoma or transitional cell carcinoma of kidney. AB - PURPOSE: We tested the discrimination ability of 2 monoclonal antibodies, mAB 90 and 2-2, in renal carcinomas, including renal cell carcinoma and transitional cell carcinoma of the kidney. MATERIALS AND METHODS: Two monoclonal antibodies raised in renal adenocarcinoma (mAB 90, IgG3 subclass and mAB 2-2, IgG1 subclass), have been generated in our laboratory by hybridoma technique. The tumor associated antigens recognized by these IgG subclass monoclonal antibodies are located in the cell membrane of tumor cells. Antibodies were purified from mice ascites through protein A-Sepharose 4B affinity column and then conjugated with fluorescein isothiocyanate fluorescence by dimethyl sulfoxide method. The binding activity of these antibodies was measured by direct immunofluorescence method and analyzed in a flow cytometer. Forty-five cases of renal cell carcinoma and 16 cases of transitional cell carcinoma of the renal pelvis were collected, and the recognition power of these 2 antibodies was tested. Frozen tumor tissues were prepared and allocated into single cell suspensions in phosphate buffered saline before adding diluted (1:10) conjugated antibodies. Irrelevant antibody and negative tumor cell lines without any reaction with these antibodies were used as background fluorescence control. RESULTS: Reactivities of mAB 90 and mAB 2-2 for renal cell carcinoma tissues were 80 and 91%, respectively. On the contrary the reactivities of mAB 90 and mAB 2-2 for transitional cell carcinoma of renal pelvis tissues were 0 and 69%, respectively. mAB 90 (+)/mAB 2-2 (-) expression pattern had 76% accurate diagnostic rate for renal cell carcinoma and mAB 90 (+)/mAB 2-2 (+) pattern had 69% accurate diagnostic rate for transitional cell carcinoma of the kidney. CONCLUSIONS: When facing pathologically ambiguous kidney tumors, the binding specificity of mAB 90 and mAB 2-2 may be useful for discrimination between renal cell carcinoma tumors and transitional cell carcinoma of the renal pelvis. PMID- 9400435 TI - Secondary ureteroscopy: results and management strategy at a referral center. AB - PURPOSE: In an era when extracorporeal shock wave lithotripsy occupies a dominant place in the treatment of urolithiasis ureteroscopy retains an important role in certain circumstances. While often a definitive procedure, ureteroscopy can be associated with potential risks and complications. The treatment of patients who have undergone a failed attempt at ureteroscopic stone retrieval or have a complication may be complex. As a tertiary care stone referral center we review our experience with performing salvage ureteroscopy following a previous unsuccessful attempt at endoscopic stone removal. MATERIALS AND METHODS: Between May 1990 and February 1996, 79 patients were referred following an unsuccessful attempt at retrograde endoscopic or basket manipulation for ureteral calculi. A retrospective review of the outcomes of these patients was conducted. Of the patients 11 presented with associated complications, which included ureteral perforation (4), intramural false passage (1) and fever or sepsis (6). Complications were managed by early establishment of urinary tract drainage by stenting or nephrostomy. Among patients without complications elective salvage ureteroscopy was performed. RESULTS: Ureteroscopy was used in 79 patients with a successful outcome (stone-free) in 75 (95%). Followup imaging with renal ultrasound or excretory urography at least 3 months after secondary ureteroscopy was available in 65 patients and showed no evidence of hydronephrosis or delayed stricture formation. CONCLUSIONS: Treating the patient who undergoes a failed attempt at ureteroscopy may be problematic and requires access to a wide array of endourological equipment. Each subsequent treatment should be individualized with consideration given to stone size, location and general health. In the presence of a ureteral injury establishment of early urinary tract drainage is essential. Following stabilization, secondary ureteroscopy can be performed yielding high stone-free rates with minimal complications. PMID- 9400436 TI - Endopyelotomy for primary ureteropelvic junction obstruction: risk factors determine the success rate. AB - PURPOSE: We prospectively assessed the feasibility, complications, and short-term and long-term results of endopyelotomy for primary ureteropelvic junction obstruction. MATERIALS AND METHODS: In 80 consecutive patients primary ureteropelvic junction obstruction was diagnosed by excretory urogram or nephrostomogram, retrograde pyelography, diuresis renography and the Whitaker test in ambiguous cases. In all patients antegrade endopyelotomy was performed with a cold knife and an indwelling stent was left for 6 weeks. At 6 and 24 months postoperatively results were assessed clinically by an excretory urogram and/or diuretic renography and later by questionnaire and ultrasound. RESULTS: The primary success rate was 89% (71 of 80 patients) after the first endopyelotomy and increased to 91% (73 of 80 patients) after 2 patients had a second endopyelotomy. After median followup of 26 months (range 1.5 to 72) 6 of the 73 initially successfully treated patients had relapse. Two were successfully re-treated by a second endopyelotomy, resulting in an overall success rate of 81% (65 of 80 patients) after 1 procedure and 86% (69 of 80 patients) after a second endopyelotomy in 4 patients. Mean preoperative pyelocaliceal volume decreased from 64 +/- 33 to 41 +/- 20 ml. (p = 0.0003) 6 months after endopyelotomy and did not change during the following 18 months. The probability of successful endopyelotomy was better in patients with a preoperative pyelocaliceal volume less than 50 ml. (87%) and worse in patients with a volume greater than 50 ml. (76%). A crossing vessel to the lower pole of the kidney causing persistent functional obstruction of the ureteropelvic junction was found in 6 of the 10 patients re-treated by open pyeloplasty (9) or nephrectomy (1). Preoperative mean renal function as determined by diuretic renography was significantly lower in patients with failed endopyelotomy than in successfully treated patients. Successfully treated patients showed no change in renal function 6 and 24 months postoperatively. CONCLUSIONS: Endopyelotomy in primary ureteropelvic junction obstruction is a safe, minimally invasive procedure with a high primary success rate and a low relapse rate. Open pyeloplasty could be avoided in 86% of our patients. Endopyelotomy is less invasive, has less functional and esthetic sequelae than open pyeloplasty and does not compromise open surgery if that becomes necessary. We recommend endopyelotomy as first line treatment for patients with primary ureteropelvic junction obstruction. PMID- 9400437 TI - Retrospective analysis of the effect of crossing vessels on successful retrograde endopyelotomy outcomes using spiral computerized tomography angiography. AB - PURPOSE: Using spiral computerized tomography (CT) angiography, we sought to evaluate the incidence of a crossing vessel in a group of adults with primary ureteropelvic junction obstruction who had previously undergone successful retrograde endopyelotomy. MATERIALS AND METHODS: A total of 16 patients who had undergone successful Acucise balloon incision endopyelotomy for ureteropelvic junction obstruction, all with followup greater than 2 years, underwent a spiral CT angiogram with intravenous contrast material to identify those with a crossing vessel. Contrast enhanced CT was performed with dual phase technique on a Somatom Plus-S CT scanner using prototype software. After 180-degree linear interpolation of the projection data, transaxial images of the affected kidney were reconstructed. In addition, at the time of the study all patients completed analog followup pain scales and quality of life assessment questionnaires. RESULTS: Among the 16 patients 6 (38%) had anterior or posterior crossing vessels based on spiral CT angiography. No patient had both types. By analog pain scale patients had 80% mean improvement in pain (range 63 to 100). CONCLUSIONS: In our series nearly 40% of patients with anterior or posterior crossing vessels had a long-term (greater than 2 years) successful outcome with retrograde endopyelotomy. Endopyelotomy continues to be our initial mode of therapy among adults with primary ureteropelvic junction obstruction. In our opinion the adverse influence of the crossing vessel is not sufficient to justify the added expense of preoperative angiography, spinal CT or endoluminal ultrasound. PMID- 9400438 TI - Endopyelotomy--a panacea for ureteropelvic junction obstruction? PMID- 9400439 TI - Ureteropelvic junction disruption secondary to blunt trauma: excretory phase imaging (delayed films) should help prevent a missed diagnosis. AB - PURPOSE: Ureteropelvic junction disruption is a rare condition which is often diagnosed after some delay. The aim of this study is to examine the current status of this entity and to determine if improvements could be made in the diagnosis. MATERIALS AND METHODS: We evaluated 5 consecutive adult cases of ureteropelvic junction disruption secondary to blunt trauma and compared the findings to those reported in literature. RESULTS: The diagnosis was delayed by at least 24 hours in 4 of the 5 cases (80%). Compared to the literature, in which most delays in diagnosis were the result of genitourinary tract imaging being omitted, most of our delays (3 cases) were a result of the initial contrast enhanced spiral (helical) computerized tomography (CT) failing to provide the diagnosis. This failure occurred because of either absence of contrast extravasation (2 cases) or only subtle extravasation (1 case), which was not recognized by the radiologist. The delay in diagnosis resulted in added morbidity in all circumstances. CONCLUSIONS: Ureteropelvic junction disruption continues to be diagnosed late in a large proportion of cases. Absence of gross contrast extravasation on nephrogram phase scanning using spiral CT may not exclude a major injury of the ureteropelvic junction. Addition of delayed CT of the kidney 5 to 8 minutes or longer after contrast material injection (during the excretory phase) may increase the probability of extravasation being demonstrated and, thus, reduce the possibility of missing a ureteropelvic junction disruption. PMID- 9400440 TI - New techniques for the administration of topical adjuvant therapy after endoscopic ablation of upper urinary tract transitional cell carcinoma. AB - PURPOSE: We evaluated the role of combining ureteroscopic tumor ablation and new methods of administering topical adjuvant therapy in select patients with transitional cell carcinoma of the upper urinary tract. MATERIALS AND METHODS: Patients with upper tract transitional cell carcinoma and indications for preserving renal function initially underwent ureteroscopic evaluation and tumor ablation. We treated 17 renal units in 13 patients. Three patients with distal ureteral lesions underwent uncomplicated adjuvant bacillus Calmette-Guerin therapy by the combination of Double-J stent placement and bladder instillations in the Trendelenburg position. In the remaining 14 renal units adjuvant topical therapy was administered by low pressure weekly instillations through a transvesical single-J stent whose curl was positioned in an upper calix. Patients were regularly followed with office flexible ureterorenoscopy under local anesthesia and cytology washings once they were confirmed to be tumor-free. RESULTS: Complete ureteroscopic tumor ablation was possible in all patients. Two sessions were needed due to tumor burden in 3 patients. There were intercurrent urinary infectious complications with Candida albicans in the 3 patients treated with bacillus Calmette-Guerin through the single-J stent, including 1 who received only 3 instillations due to persistent unexplained fevers but who remains disease-free at 24 months. In 2 of the 17 renal units multifocal tumor recurred within 12 months, which was treated with nephroureterectomy. At short followup (mean 14.6 months) 15 renal units have been preserved and remain tumor free. CONCLUSIONS: The new techniques of administering adjuvant topical agents for upper tract transitional cell carcinoma after ureteroscopic tumor ablation in select cases described provide a useful and simple alternative to the percutaneous method preferred in the past. Short-term responses have been favorable but the true value of adjuvant therapy remains uncertain at present. The 2 recurrences within 12 months of treatment were readily detected by outpatient ureterorenoscopy with the patient under local anesthesia using 7.5F flexible endoscopes. PMID- 9400441 TI - Isolated retrovesical and extrarenal retroperitoneal hydatidosis: clinical study of 10 cases and literature review. AB - PURPOSE: Isolated extrarenal retroperitoneal echinococcal cyst is a rare manifestation of hydatid disease. We review the clinical findings of a series of isolated retrovesical and retroperitoneal hydatid cysts in an endemic area, with special emphasis on diagnostic pitfalls. MATERIALS AND METHODS: A retrospective 15-year review in a rural area of central Spain (600,000 population), with a global incidence of hydatidosis of 10 new cases per 100,000 population per year, revealed 10 patients (0.1 cases per 100,000 per year) with surgically treated primary extrarenal retroperitoneal echinococcosis and absence of other hydatid cysts in any organ. Clinical, radiological and laboratory data are analyzed. The literature of the last 15 years on hydatid disease with a primary retroperitoneal and retrovesical location is reviewed. RESULTS: A total of 42 cases of isolated retrovesical (25) and retroperitoneal (17) hydatidosis was compiled. Male predominance of 2.5:1 was noted and age ranged from 8 to 79 years (mean 42.2 +/- 5.4). The most frequent presentation was a palpable mass in 29% of the cases followed by flank pain in 24%, frequency in 17% and urinary retention in 14% of the cases. Hydatiduria of the urinary tract was present in 9.5% of the cases. Different serological investigations were performed in 45% of the cases with positive results in 68%. The lesion had been surgically removed in all cases. Therapy and followup of each patient were analyzed. CONCLUSIONS: An isolated retrovesical, retroperitoneal or even retrocrural cyst can be the unique manifestation of hydatid disease. Although difficult, preoperative diagnosis is desirable for the selection of a surgical approach and prevention of allergic reactions and operative spillage. A diagnostic algorithm and several therapeutic guidelines are proposed. PMID- 9400442 TI - Bladder petechiae after cystoscopy and hydrodistension in men diagnosed with prostate pain. AB - PURPOSE: We determined if men with prostate pain syndromes have petechiae in the bladder after hydrodistension. MATERIALS AND METHODS: A total of 60 men with the diagnosis of prostate pain and without bacteriuria underwent cystoscopy and hydrodistension under a general or regional anesthetic. RESULTS: Of the 60 men 35 (58%) had moderate to severe petechiae similar in appearance to women with interstitial cystitis after hydrodistension. Men with moderate to severe bladder petechiae had fewer leukocytes in expressed prostatic secretions, smaller bladder capacities and less often testicular pain than men with more normal appearing bladders after hydrodistension. Symptomatic improvement 2 to 6 weeks after hydrodistension was more common in men with moderate to severe petechiae than in those with fewer petechiae. Absence of rectal pain predicted symptomatic improvement after hydrodistension. CONCLUSIONS: We suggest that bladder petechiae, and possibly interstitial cystitis or a related condition, may be more frequently associated with prostate pain syndromes in men than previously appreciated. PMID- 9400443 TI - Lack of evidence for a role of human papillomaviruses in transitional cell carcinoma of the bladder. AB - PURPOSE: In view of the conflicting results reported in the literature, we assessed the involvement of human papillomaviruses (HPV) in the development of transitional cell carcinoma of the bladder. MATERIALS AND METHODS: A total of 58 bladder papillomatous proliferations was histologically examined and analyzed for the presence of HPV deoxyribonucleic acid (DNA) sequences by Southern blot hybridization and the polymerase chain reaction (PCR) method. RESULTS: Typical features of condyloma acuminatum were observed in the bladder specimen of a patient with urethral condylomatosis. Of the specimens 57 had histological features of transitional cell carcinoma but no known signs of HPV infection. HPV 6 DNA was detected in the condylomatous tumor. However, no HPV DNA was detected in the 57 bladder cancers by Southern blot hybridization and polymerase chain reaction. CONCLUSIONS: These findings do not support an etiological role of HPV in the development of transitional cell bladder cancer. PMID- 9400444 TI - The influence of the level of lamina propria invasion and the prevalence of p53 nuclear accumulation on survival in stage T1 transitional cell bladder cancer. AB - PURPOSE: We assessed the influence of the level of lamina propria invasion and the prevalence of p53 nuclear immunoreactivity on the survival of patients with stage T1 transitional cell bladder cancer. MATERIALS AND METHODS: All patients presenting with stage T1 bladder cancer were prospectively and routinely grouped according to the level of lamina propria invasion. Invasion of the tumor stalk was defined as stage T1a, invasion of the lamina propria proper superficial to the level of muscularis mucosa as stage T1b and into or deeper than the muscularis mucosa as stage T1c. The p53 nuclear immunoreactivity was determined with antibody PAB 1801. RESULTS: The study comprised 143 patients including 31 (22%) with stage T1a disease, 60 (42%) with stage T1b and 52 (36%) with stage T1c. Mean patient age was 67 years (range 38 to 92) and mean followup was 4.7 years (range 2.4 to 9.7). Tumor grade related to the depth of lamina propria invasion (p < 0.05) and the prevalence of dysplasia in random mucosal biopsies was higher in stage T1b and T1c tumors than in stage T1a. Of all tumors 42% expressed p53 nuclear reactivity which correlated with tumor grade (p < 0.05). Also the prevalence of nuclear p53 was higher in stages T1b and T1c compared with T1a disease. Of the patients 115 were treated with transurethral resection alone and 28 underwent radical cystectomy. Overall survival was 60.1%. Age was the only independent predictor of survival in patients older than 75 years. For patients up to 75 years old survival related to age, level of lamina propria invasion and presence of p53 nuclear accumulation. For this subpopulation overall survival was 67%, and 79% for stage T1a, 70% for stage T1b and 57% for stage T1c (p < 0.05). Survival was higher in patients with p53 negative (73%) than in those with p53 positive (61%) tumors (p < 0.05). A multivariate analysis of the influence of lamina propria invasion and nuclear p53 status on survival histology was found to be the only independent predictor of survival. CONCLUSIONS: Immediate radical cystectomy should be considered for patients with stage T1c tumors and for some patients with stage T1b disease, particularly those with tumors expressing p53 nuclear reactivity and with dysplasia in the random mucosal biopsies. PMID- 9400446 TI - Transurethral ultrasound: evaluation of anatomy and function of the rhabdosphincter of the male urethra. AB - PURPOSE: A combined anatomic-sonographic study was undertaken to investigate whether the anatomical arrangement and the contractions of the rhabdosphincter of the male urethra could be visualized by transurethral ultrasound. Furthermore, this new technique was compared with standard urodynamic tests. MATERIALS AND METHODS: In 7 cadavers transurethral ultrasound was performed to define sono morphological criteria of the rhabdosphincter, and the sonographic pictures were then compared to histological sections. In 48 patients the rhabdosphincter of the male urethra was investigated by transurethral ultrasound and urodynamic techniques. Of these patients 40 were completely continent after radical prostatectomy and 8 presented with urinary stress incontinence after transurethral resection of the prostate or radical prostatectomy. The decrease of the distance between the rhabdosphincter and the transducer during contraction served as quantitative parameter for the contractility of the muscle. RESULTS: The anatomical arrangement and contractions of the rhabdosphincter loop could be clearly visualized on transurethral ultrasound (during contraction the rhabdosphincter retracts the urethra, pulling it towards the rectum). Ultrasound showed scars in 3 patients with postoperative urinary stress incontinence, thinning of the muscle in 3 complete atrophy of the rhabdosphincter in 2 and minimal contractions of the rhabdosphincter in 1. Urethral closure pressures were decreased and decrease in rhabdosphincter-transducer distance was statistically significantly decreased in the incontinent patients. CONCLUSIONS: Our sono morphological data and anatomical histological results strongly suggest that the rhabdosphincter constitutes the main component of the continence mechanism in post-prostatectomy patients. Unlike urethral pressure profiles, which can only reveal zones of higher intraluminal pressure between the bladder and the penile urethra, transurethral ultrasound is highly specific for measurement of the function of the rhabdosphincter. PMID- 9400445 TI - Feasibility of transurethral resection for muscle infiltrating carcinoma of the bladder: long-term followup of a prospective study. AB - PURPOSE: We analyzed the long-term results of radical transurethral resection for the treatment of a large series of patients with muscle infiltrating bladder cancer entered into a prospective study to determine progression predictive factors. MATERIALS AND METHODS: The study included 133 patients with invasive bladder cancer treated by radical transurethral resection who had negative biopsies of the muscle layer of the tumor bed. Followup was more than 5 years for all subjects and more than 10 years in 59 (44.4%). A comparative nonrandomized study was performed of a control group of 76 patients with invasive pathological stage pT2-3a, N0-3 bladder cancer treated by cystectomy. In those patients treated by radical transurethral resection univariate and multivariate analyses were performed to establish clinical progression predictive factors. RESULTS: At 5 and 10 years of followup cause specific survival rates were 80.5 and 74.5%, and bladder preservation rates were 82.7 and 79.6%, respectively. No significant difference was noted in terms of cause specific survival, with respect to the control group. The initial presence of associated bladder carcinoma in situ was the only independent progression predictive factor. CONCLUSIONS: For patients with invasive bladder cancer radical transurethral resection is justified when the tumor is clinically limited to the muscular layer and when all biopsies of the periphery and depth of the tumor bed show muscular tissue negative for tumor cells. Patients with initial associated bladder carcinoma in situ should not be excluded from this treatment but endovesical bacillus Calmette-Guerin immunotherapy should be administered and a closer followup is recommended. PMID- 9400447 TI - A followup on transurethral collagen injection therapy for urinary incontinence. AB - PURPOSE: Transurethral collagen injection therapy has been used successfully in treating stress urinary incontinence due to intrinsic sphincter deficiency since United States Food and Drug Administration approval in October 1993. MATERIALS AND METHODS: Telephone interview and chart review were performed on 139 women with intrinsic sphincter deficiency documented using video urodynamics, of whom 73% had grade 3 incontinence (leakage without effort). Median followup was 18 months (range 6 to 36). Median patient age was 72 years. RESULTS: A total of 103 patients (74%) was substantially improved after collagen therapy, 29 (20%) were improved and 7 had no improvement. Of the substantially improved group 72% obtained continence after 2 or fewer injections. Of the patients 11% required a "booster" injection more than 6 months after initial treatment. Complications, such as hematuria, urinary tract infections or transient urinary retention, were rare. CONCLUSIONS: Our results confirm the safety and efficacy of transurethral collagen. Once continence is achieved further collagen therapy is rarely necessary. PMID- 9400448 TI - Hemodynamic patterns of pharmacologically induced erection: evaluation by color Doppler sonography. AB - PURPOSE: Penile erection is achieved through hemodynamic mechanisms that can be assessed best with color flow imaging and Doppler waveform analysis. We performed dynamic studies using computer assisted analysis to assess the hemodynamic patterns of pharmacologically induced erection. MATERIALS AND METHODS: A total of 73 color Doppler ultrasound studies was performed in 66 patients with erectile dysfunction. Various blood flow parameters, including peak systolic velocity, end diastolic velocity, mean flow rate, resistive index and artery diameter, were observed continuously and recorded frequently for about 30 minutes after intracorporeal injection of papaverine/phentolamine/prostaglandin E1 mixture. A computerized Doppler waveform analysis of 3 curves or greater was performed for each recording to minimize error. A second injection was administered if the first injection failed to induce a rigid erection. Status of the erection was observed and recorded throughout the study. A computerized graph was generated for each corpus. RESULTS: After intracorporeal injection the time to reach normal or peak velocity varied from 1 to 24 minutes. Among 146 corpus units in 73 color Doppler ultrasound studies we observed the following hemodynamic patterns: I normal maximal peak systolic velocity (35 cm. per second or greater), sustained; Ia-end diastolic velocity 0 or less with complete erection response (19 units); Ib-end diastolic velocity greater than 0 or incomplete erection response (14 units); II-normal maximal peak systolic velocity (35 cm. per second or greater), transient; IIa-end diastolic velocity 0 or less with complete erection response (21 units); IIb-end diastolic velocity greater than 0 or incomplete erection response (12 units); III-borderline maximal peak systolic velocity (30 to 35 cm. per second); IIIa-end diastolic velocity 0 or less with complete erection response (10 units); IIIb-end diastolic velocity greater than 0 or incomplete erection response (8 units); IV-low maximal peak systolic velocity (less than 30 cm. per second); IVa-end diastolic velocity 0 or less with complete erection response (24 units); and IVb-end diastolic velocity greater than 0 or incomplete erection response (38 units). CONCLUSIONS: Erection is a complex and dynamic process. A new classification of hemodynamic patterns is presented that aids in assessing and interpreting more thoroughly blood flow parameters to stratify more precisely the hemodynamic patterns of erectile dysfunction. PMID- 9400449 TI - Genital plus audiovisual sexual stimulation following intracavernous vasoactive injection versus re-dosing for erectile dysfunction--results of a prospective study. AB - PURPOSE: We assessed whether re-dosing of a vasoactive agent or the combination of a vasoactive injection and genital plus audiovisual sexual stimulation caused the greatest erectile effect to determine which of the 2 procedures would be better for dynamic penile color Doppler sonography in patients with erectile dysfunction. MATERIALS AND METHODS: A total of 20 consecutive patients with erectile dysfunction underwent 2 sessions under real-time RigiScan* recording of penile erection. Session 1 consisted of adaptation in 10 minutes, intracavernous injection of 10 micrograms. alprostadil in 10 minutes and re-dosing of 10 micrograms. alprostadil in 10 minutes. Session 2 consisted of adaptation in 10 minutes, injection of 10 micrograms. alprostadil in 10 minutes and genital plus audiovisual sexual stimulation in 10 minutes. The total duration of each session was 30 minutes. The order of the 2 sessions was randomly assigned with a week interval between each session. RESULTS: Re-dosing and genital plus audiovisual sexual stimulation caused a significant increase in erectile response compared to the result seen after the first injection (re-dosing p < 0.05, injection plus stimulation p < 0.01). However, erectile response after the genital stimulation session was significantly greater than that after re-dosing (p < 0.01). An erection comparable to the greatest spontaneous erection reported by the patient was much more frequently achieved after genital stimulation than after the re dosing session (p < 0.01). CONCLUSIONS: The combination of injection and stimulation caused a significantly greater erectile response than re-dosing. We suggest that the former should always be used during color Doppler sonography to optimize the accuracy of the test. Re-dosing is suggested when an incomplete erectile response occurs after the injection plus stimulation phase. PMID- 9400450 TI - Double-blind multicenter study comparing alprostadil alpha-cyclodextrin with moxisylyte chlorhydrate in patients with chronic erectile dysfunction. AB - PURPOSE: We compared the efficacy and safety of alprostadil alpha-cyclodextrin and moxisylyte chlorhydrate to induce erections adequate for sexual intercourse in a prospective, randomized, parallel double-blind study in patients with erectile dysfunction. MATERIALS AND METHODS: A total of 156 men with erectile dysfunction due to organic, nonorganic and mixed origin was randomized into 2 parallel treatment groups receiving titrations of an individual optimum dose of alprostadil alpha-cyclodextrin or moxisylyte chlorhydrate. Erectile response was measured by the buckling test. A positive test was associated with axial erection rigidity that did not buckle/deform to 1.0 kg. load. The buckling test was repeated every 10 minutes for up to 60 minutes. RESULTS: A total of 56 patients (75%) in the alprostadil alpha-cyclodextrin group and 32 patients (40%) in the moxisylyte chlorhydrate group responded with at least 1 positive buckling test during the office period. Investigators assessed erections after alprostadil alpha-cyclodextrin to be adequate for sexual intercourse in 61 patients (81%) compared to 37 patients (46%) after moxisylyte chlorhydrate. All efficacy parameters in office reached statistical significance of p < 0.001 in favor of alprostadil alpha-cyclodextrin. During self-injection therapy at home 58 patients (85%) reported at least 1 rigid erection after alprostadil alpha-cyclodextrin compared to 37 patients (61%) after moxisylyte chlorhydrate. Patient and partner opinion of treatment achieved statistically significantly better scores in the alprostadil alpha-cyclodextrin group compared to the moxisylyte chlorhydrate group. CONCLUSIONS: Alprostadil alpha-cyclodextrin is significantly more effective than moxisylyte chlorhydrate in producing full penile rigidity in office and at home. Injection related side effects occur with the same frequency but moxisylyte results in more systemic side effects and alprostadil results in more painful and prolonged erections. Patients and partners are significantly more satisfied with alprostadil alpha-cyclodextrin. PMID- 9400451 TI - After the gold rush--advancing research based care for patients with erectile dysfunction. PMID- 9400452 TI - Therapeutic effects of high dose yohimbine hydrochloride on organic erectile dysfunction. AB - PURPOSE: We evaluated men with organic erectile dysfunction treated with placebo and high dose oral yohimbine hydrochloride. MATERIALS AND METHODS: We selected 22 patients with organic erectile dysfunction (mean age 58 years) for treatment in the andrology outpatient clinic. These patients had been previously undergone neurological, vascular, hormonal and psychological testing, and were treated during an equal period of 30 days with placebo and daily single dose oral 100 mg. yohimbine. The response to treatment was evaluated via a questionnaire that comprised the outcome items of complete--normal penile rigidity enabling vaginal penetration, partial--erection improved but not sufficiently for appropriate vaginal penetration, none--no improvement and worse--erection deteriorated. The patients consented to treatment after being told of the severe adverse effects that might occur. RESULTS: The most common side effects were anxiety, increase in cardiac frequency, increased urinary output and headache but in no case was treatment discontinued. Of the patients 3 (13.6%) and 12 (54.5%) reported complete or partial response to treatment, respectively. However, statistical analysis disclosed no significant difference when yohimbine was compared to placebo (p < 0.05). CONCLUSIONS: Oral 100 mg. single dose daily yohimbine promotes no improvement in patients with organic erectile dysfunction. PMID- 9400453 TI - Management of severe corporeal fibrosis with implantation of prosthesis via a transverse scrotal approach. AB - PURPOSE: We reviewed our experience in the management of severe corporeal fibrosis with placement of the AMS 700 CXM* prosthesis to determine the efficacy of this approach. MATERIALS AND METHODS: The records of 26 men with severe corporeal fibrosis who underwent placement of the AMS 700 CXM prosthesis via a transverse scrotal approach between August 1991 and July 1996 were reviewed. RESULTS: In all cases the AMS 700 CXM prosthesis was successfully implanted with primary closure of the tunica albuginea, although 2 patients required extended corporotomies. Followup data were available on all 26 men. At a mean followup of 22.5 months (range 3 to 63) 24 of the 26 men had a functional device (92%). One patient required explantation for infection and 1 underwent explantation for cylinder cross-over. CONCLUSIONS: Implantation of the AMS 700 CXM prosthesis in patients with severe corporeal fibrosis produced good results at approximately 2 years of followup. PMID- 9400454 TI - Use of a prefabricated tunica vaginalis fascia flap to reconstruct the tunica albuginea after recurrent penile prosthesis extrusion. AB - PURPOSE: Although a penile prosthesis usually perforates into the urethra, it can extrude through the glans or corporeal shaft. Various materials have been used to reconstruct tunica albuginea but no method of repair has been satisfactory in such difficult cases. Repair of the weakened tunica albuginea should ideally be performed with autogenous tissues. Inasmuch as the scarred tissue bed is inadequate to ensure graft survival and no local flaps are available for this purpose, prefabrication of a local flap has been designed. MATERIALS AND METHODS: We present 2 cases in which the distal corpus was reconstructed with a prefabricated tunica vaginalis fascia flap. The first stage involves grafting rectus fascia onto the external tunica vaginalis of the testicle. At the second stage the prefabricated tunica vaginalis fascia flap is transposed to the distal corpus, placing it as a buttress between the cylinder and urethra medially or between the cylinder and thin lateral and distal tunica albuginea. The flap also replaces part of the tunica albuginea. RESULTS: In both patients repair of the tunica albuginea was successful and each has a functioning inflatable penile prosthesis at 2 1/2 1 1/2 years postoperatively, respectively. CONCLUSIONS: Reconstruction of the weak tunica albuginea with a prefabricated tunica vaginalis fascia flap is an excellent procedure in these difficult cases. PMID- 9400455 TI - Teratoma with malignant transformation: diverse malignant histologies arising in men with germ cell tumors. AB - PURPOSE: Teratoma with malignant transformation refers to a form of germ cell tumor in which a somatic teratomatous component becomes morphologically malignant and develops aggressive growth. We evaluated the spectrum of histologies, chromosomal abnormalities and clinical outcome in patients with teratoma with malignant transformation. MATERIALS AND METHODS: We identified 46 patients with germ cell tumor meeting morphologic criteria for malignant transformation. Histology, disease extent and treatment were correlated with survival. Tumors in 12 patients were studied by conventional cytogenetics or molecular genetic techniques for the isochromosome 12p [i(12p)], a marker for germ cell tumor, as well as other chromosomal abnormalities. RESULTS: The site of first detection of malignant transformation occurred in the primary tumor of 21 cases (44%), at a metastatic site in 20 (43%) and in both sites in 5 (10%). Sarcoma was the most frequent histology, identified in 29 patients (63%) with rhabdomyosarcoma the most common subtype. Seventeen tumors (37%) contained a solid tumor histology other than sarcoma, with adenocarcinoma and primitive neuroectodermal tumor as the most common histologies. Four patients with mediastinal germ cell tumor containing sarcoma also had hematological malignancies, including a focus of nonHodgkin's lymphoma in the mediastinal primary tumor (1) and nonlymphocytic leukemia in spleen or bone marrow (3). Patients who had teratoma with malignant transformation components confined to the testis or retroperitoneum completely resected experienced a longer survival than those with distant metastases or incompletely resected tumors (p = 0.003). Chromosomal abnormalities associated with germ cell tumor (i[12p]) were identified in 11 of 12 tumors containing adenocarcinoma, primitive neuroectodermal tumor, sarcoma and leukemia. In addition to i (12p), chromosomal rearrangements characteristic of the transformed histology were detected in 4 tumors. CONCLUSIONS: A variety of nongerm cell histologies, including sarcoma, adenocarcinoma, primitive neuroectodermal tumor and leukemia, may occur in association with germ cell tumor. Chromosomal abnormalities in these tumors include i (12p), reflecting germ cell tumor clonality, as well as chromosomal abnormalities associated with the transformed histology. These tumors do not respond like germ cell tumor to cisplatin containing chemotherapy regimens. Treatment should be tailored according to that used in standard management of the transformed histology, and surgical resection is the mainstay of therapy. PMID- 9400456 TI - Microsurgical repair of iatrogenic injury to the vas deferens. AB - PURPOSE: We determined the incidence of iatrogenic injuries to the vas deferens at a tertiary care university infertility center and the results of surgical repair. MATERIALS AND METHODS: Records of 472 patients surgically explored for obstructive azoospermia between 1984 to 1996 were reviewed. Enrollment criteria included history of inguinal, pelvic and scrotal (other than vasectomy) surgery. Conventional ipsilateral and crossover vasovasostomies and vasoepididymostomies were performed. Patency rate was defined as presence of complete sperm with tails in a postoperative semen analysis. Followup included a minimum of 2 semen analyses. Only naturally conceived pregnancies were included. RESULTS: Of 472 patients 34 (7.2%) had an iatrogenic injury to the vas deferens with a mean obstruction interval of 20.5 +/- 1.9 years. Mean patient age was 36.7 +/- 1.8 years. Iatrogenic injury to the vas deferens was secondary to bilateral inguinal hernia repair in 19 patients, unilateral hernia repair in 11, renal transplantation in 2, appendectomy in 1 and spermatocelectomy in 1. Pediatric inguinal hernia repair was the most common etiology of the vasal injury (20 patients), followed by adult inguinal hernia repair (10). A total of 36 microsurgical reconstructive procedures were performed, including 20 ipsilateral and 16 crossed vasovasostomies and vasoepididymostomies. There were 26 patients (29 procedures) available for followup (mean 21.0 +/- 3.7 months). Total patency rate per procedure was 65% and pregnancy rate was 39%. Patency and pregnancy rates per conventional ipsilateral procedures were 62.5 and 35.7% and per crossover procedures 64.2 and 42.8%, respectively. CONCLUSIONS: Pediatric inguinal hernia repair is the most common cause of iatrogenic injury to the vas deferens. Results of treatment of iatrogenic injury to the vas deferens are somewhat lower than for patients with obstructive azoospermia due to vasectomy. Iatrogenic injuries are associated with longer vasal defects, impaired blood supply and longer obstructive intervals frequently resulting in secondary epididymal obstruction. Crossover reconstruction is particularly useful when contralateral testicular atrophy is present. Intraoperatively aspirated sperm should be cryopreserved for later use in case the reconstruction fails. PMID- 9400457 TI - Three-dimensional stereotactic posterior ischiorectal space computerized tomography guided brachytherapy of prostate cancer: a preliminary report. AB - PURPOSE: A 3-dimensional (D) stereotactic posterior ischiorectal space computerized tomography (CT) guided approach is presented for brachytherapy of localized prostate adenocarcinoma. MATERIALS AND METHODS: During the last 2 years 130 patients 49 to 90 years old (median age 71) with clinical stage A, B or C adenocarcinoma have been treated by this method. The initial prostate specific antigen profile was range 0.9 to 143 ng./ml., mean, 16.25 and median 13.0. Range of initial prostatic volume was 30 to 156 cm.3, with a (median 62 and mean 65). Of the patients 15% had signs and symptoms of urinary obstruction, that is with residual urine greater than 100 cc and significant nocturia and frequency. Transurethral resection of the prostate defects were present in 20% of the patients. Volume and treatment planning is performed by CT. Placement of the after loading needles is accomplished with a 3-D stereotactic system mounted on a CT table. The prescribed dose is 12,000 cGy. for 103Palladium seeds and 16,000 for 125I. The dosage is achieved by spacing the after loading needles 10 mm. apart with the seeds averaging 10 mm. apart from center to center. RESULTS: Prostate specific antigen levels decreased to less than 2 ng./ml. in 95% of the patients including those at high risk 6 to 24 months after the procedure. Except for treatment related transient symptoms of urethritis and proctitis, there have been no complications. No patients had incontinence, acute infection, hemorrhage or radiation damage to the rectum. No patients required post-implant transurethral resection of the prostate. There was significant clinical improvement in patients with obstructive uropathy. CONCLUSIONS: The 3-D stereotactic CT guided posterior ischiorectal space approach for brachytherapy is not limited by prostate size, transurethral prostatic resection defects or public arch interference, and it allows for needle verification and correction if necessary. Initial clinical and biochemical results in patients treated with this method are promising. PMID- 9400458 TI - Kaposi's sarcoma associated herpesvirus deoxyribonucleic acid sequences: lack of detection in prostatic tissue of human immunodeficiency virus-negative immunocompetent adults. AB - PURPOSE: A recent study argued that Kaposi's sarcoma associated herpesvirus is ubiquitous, as reflected by the frequent detection of its deoxyribonucleic acid (DNA) sequences in the prostatic tissue of healthy Italian men. Because these findings are discordant with serological data, our objective was to reassess the prevalence of this virus in prostate tissue from immunocompetent men. MATERIALS AND METHODS: Normal tissue samples from 45 human immunodeficiency virus-negative men undergoing radical prostatectomy for prostate carcinoma were snap frozen. DNA was extracted from normal noncancerous tissue. DNA was confirmed to be polymerase chain reaction amplifiable. Then, using multiple primer sets, extracted prostatic DNA was blinded, polymerase chain reaction amplified and screened for Kaposi's sarcoma herpesvirus by Southern blot. RESULTS: Kaposi's sarcoma herpesvirus DNA sequences were not detected in either the 45 prostatic DNA or in blinded internal negative controls, but they were consistently identified in all positive internal controls. CONCLUSIONS: Since Kaposi's sarcoma herpesvirus was undetectable in any of the prostatic samples despite high assay sensitivity, it is unlikely that the prostate serves as a reservoir for this virus in immunocompetent North American men. These findings also suggest that Kaposi's sarcoma herpesvirus is not ubiquitous. PMID- 9400459 TI - High dose bicalutamide for androgen independent prostate cancer: effect of prior hormonal therapy. AB - PURPOSE: A pilot study of the antiandrogen bicalutamide at 150 mg. a day for androgen independent prostate cancer was performed. This study was based on the possibility that androgen independent cases might display responses to additional hormonal agents. MATERIALS AND METHODS: The study included 31 androgen independent cases with an increasing prostate specific antigen (PSA) and progressive disease. PSA measurements were used as the primary method of assessing response. However, PSA decline was also correlated with clinical status. RESULTS: Seven patients demonstrated PSA declines of greater than 50% for 2 months or more, for an overall response rate of 22.5%. Responses were observed almost exclusively in patients treated with long-term flutamide as part of a complete androgen blockade regimen (43% response rate) in contrast to patients treated with androgen deprivation without flutamide (6% response rate). Of the 7 PSA responding patients bicalutamide resulted in a significant improvement in performance status and a decrease in analgesic requirement in 4 and 3 remained asymptomatic. Bicalutamide at 150 mg. a day was well tolerated, with the most frequent side effect being mild exacerbation of hot flashes. CONCLUSIONS: Bicalutamide at this dose is modestly effective for some patients with androgen independent prostate cancer, particularly for those previously treated with long term flutamide. This study indicates that previous antiandrogen therapy alters the response to subsequent hormonal agents. PMID- 9400460 TI - Bupivacaine infiltration into the neurovascular bundle of the prostatic nerve does not improve postoperative pain or recovery following transvesical prostatectomy. AB - PURPOSE: We assessed the effect of intraoperative bupivacaine infiltration into the neurovascular bundle of the prostatic nerve on postoperative pain and patient outcome. MATERIALS AND METHODS: The study included 40 American Society of Anesthesiologists physical status I to III patients undergoing transvesical prostatectomy. Following surgical resection of the prostate the neurovascular bundle of the prostatic nerve was infiltrated with either 10 ml. bupivacaine 0.5% or saline. Postoperative pain intensity was assessed using a patient generated 100 mm. visual analog scale and a patient controlled analgesia device. Additional analgesic requirements, time to ambulation, length of hospitalization and return to normal activity were also recorded. RESULTS: There were no differences in visual analog scale for pain, patient controlled analgesia demands or actual morphine delivered. Similarly, saline versus bupivacaine infiltration did not influence ambulation time (21.3 +/- 2.7 versus 25.0 +/- 11.8 hours, respectively), length of hospitalization (7.06 +/- 0.8 versus 7.11 +/- 0.6 days, respectively), return to normal activity (14.4 +/- 8.8 versus 14.2 +/- 8.2 days, respectively) or patient satisfaction. On postoperative days 1 and 2 more patients in the saline treatment group requested additional oral analgesia compared to the bupivacaine treatment group. However, no statistical difference was demonstrated. CONCLUSIONS: Following transvesical prostatectomy, prostatic nerve blockade has no beneficial effects on postoperative pain or patient outcome. PMID- 9400461 TI - Assessment of patient preferences among men with prostate cancer. AB - PURPOSE: We developed a self-administered paper based instrument to assess patient preferences quantified as utilities for common outcomes associated with the management of prostate cancer. MATERIALS AND METHODS: A total of 50 patients was invited to test a self-administered paper based instrument designed to assess preferences for health outcomes associated with the management of localized prostate cancer. The 50 patients were selected from a group of 625 randomly identified men with prostate cancer who responded to a survey instrument designed to assess health related quality of life. The 50 patients selected for this pilot project were chosen because of the wide range of responses to the quality of life survey. Patient utilities were assessed for the 5 health states of overall quality of life, problems related to prostate cancer, and problems related to urinary, bowel and sexual dysfunction. RESULTS: Patients were able to complete the assigned tasks. The self-administered instrument had high test-retest reliability. In addition results obtained from this instrument showed a correlation with results obtained from assessments using other instruments, including an analog scale, a computer based system known as U-Titer, a quality of life survey and the Health Utility Index:3. CONCLUSIONS: A self-administered paper based instrument can be used to assess patient utilities for health states associated with prostate cancer management. Results from the instrument tested appear to be reliable and valid, and are comparable to those obtained from other assessment techniques. A self-administered paper based instrument has distinct advantages when conducting large survey studies because it can be incorporated at relatively low cost. PMID- 9400462 TI - Adjuvant radiation therapy does not cause urinary incontinence after radical prostatectomy: results of a prospective randomized study. AB - PURPOSE: We analyzed the potential influence of adjuvant radiotherapy on urinary continence after radical prostatectomy. MATERIALS AND METHODS: A total of 100 patients with N0M0 prostate cancer randomized in a prospective study on postoperative radiotherapy for locally advanced disease (positive surgical margin, capsular perforation and/or seminal vesicle infiltration) were studied. Objective pad weighing tests corroborated by direct personal interviews were used to evaluate urinary continence at regular postoperative intervals. RESULTS: Of the patients 48 received 60 Gy. external radiotherapy with 18 MV photon beams between 12 and 16 weeks postoperatively, and 52 were followed expectantly. Risk factors were similar in both groups. With a mean followup of 24 months, no difference in complete urinary continence was observed. Of the irradiated group 77% and of the surveillance group 83% were totally dry. The fate of the bladder neck had no significant influence on final continence status, although there was a trend for faster recovery when the bladder neck was preserved. CONCLUSIONS: In this prospective randomized study 60 Gy. external radiation therapy administered between 3 and 4 months after radical prostatectomy for pathologically locally advanced prostate cancer had no significant influence on urinary continence. PMID- 9400463 TI - Radical retropubic prostatectomy outcomes at a community hospital. AB - PURPOSE: We reviewed a 6-year experience performing radical retropubic prostatectomy at 2 community hospitals (for-profit and not-for-profit) to assess outcomes and to compare them to the published literature reflecting outcomes from major academic hospitals. MATERIALS AND METHODS: Charts of 116 patients who underwent radical retropubic prostatectomy (nerve sparing in select cases) between 1990 and 1996 were reviewed for clinical and pathological outcomes as well as hospital charges. Subjective patient reports of urinary continence, potency and satisfaction were evaluated postoperatively. RESULTS: Average patient age was 66.6 years and average preoperative prostate specific antigen level was 9.6 ng./ml. Of the patients 43% had T1c disease, 63% pT2 and 37% pT3. Positive margins were present in 17.2% of the specimens and 66% of the patients had Gleason scores of 5 and 6. No deaths occurred. Major complications occurred in 5.4% of patients and included deep venous thrombosis (1.8%), pulmonary embolism (1.8%), rectal injury requiring ileostomy (0.9%) and fascial dehiscence (0.9%). Mean blood loss was 872 cc and mean blood transfusion rate was 1.7 units (almost exclusively autologous blood). Hospital charges decreased at the not-for-profit hospital to $13,233 in 1996 from $17,743 in 1990 to 1995, whereas charges increased at the for-profit hospital to $25,979 in 1996 from $24,481 in 1990 to 1995. Mean length of stay decreased from 5 days in 1990 to 1995 to 3 days in 1996. Of the patients 80% were totally continent (pad-free), 12% wore a protective pad once per day for minimal incontinence and 8% wore 2 or more pads. Of the men who were potent preoperatively 18% retained potency and 46% remained sexually active postoperatively either spontaneously or with assistance. Of the patients 84% were satisfied with surgical outcomes, 11% were somewhat satisfied and 5% were dissatisfied. CONCLUSIONS: Radical retropubic prostatectomy can be performed safely and with acceptable clinical and pathological outcomes at a community hospital. Impotence continues to be one of the most bothersome morbidities, particularly in older men. Increasing cost awareness, coincident with the proliferation of managed care, has led to reductions in length of hospital stay and charges at certain hospitals. PMID- 9400464 TI - Prostate cancer outcomes studies--the pathway to improvement. PMID- 9400465 TI - Long-term results of radiation therapy for prostate cancer recurrence following radical prostatectomy. AB - PURPOSE: Following radical prostatectomy, radiation therapy may be beneficial in select patients with isolated local recurrence. Pathological stage, Gleason score and the timing of prostate specific antigen (PSA) elevation are useful in distinguishing men with local recurrence from those with distant metastases. We test the ability of these criteria to predict long-term suppression of PSA recurrence following post-prostatectomy radiation therapy. MATERIALS AND METHODS: Of 1,699 men treated with radical prostatectomy from 1982 to 1995, 82 with an isolated PSA elevation or local recurrence following surgery underwent radiation therapy to the prostatic bed and were followed for at least 2 years. No patient had evidence of metastases at the time of radiation. RESULTS: Of the men 17 (21%) had an undetectable PSA (less than 0.2 ng./ml.) for 2 or greater years following radiation. The 5-year actuarial PSA recurrence-free rate after radiation was 10%. PSA remained at undetectable levels for 2 or greater years in no patients with Gleason score 8 or greater (12 cases), positive seminal vesicles (12) or positive lymph nodes (3), and in only 1 of 16 men (6%) who had a PSA recurrence within 1 year of prostatectomy. As the interval to PSA recurrence increased, the likelihood of responding to radiotherapy increased to 44% if initial disease detection occurred 5 or more years after prostatectomy. There was no demonstrated advantage to radiating men with an isolated PSA elevation before a documented local recurrence. CONCLUSIONS: Patients with Gleason score 8 or greater, positive seminal vesicles or lymph nodes, or a PSA recurrence within the first year following surgery rarely benefit from radiation therapy. As the interval to PSA recurrence increases, the likelihood of responding to radiation therapy increases substantially. These parameters are useful in the selection of patients with prostate cancer recurrences who are likely to benefit from radiation to the prostatic bed. PMID- 9400466 TI - The use of 2 ipsilateral ureteral stents for relief of ureteral obstruction from extrinsic compression. AB - PURPOSE: We present our early experience with the novel approach of placing 2 parallel stents simultaneously in extrinsically obstructed ureters in which single stents had failed. The increased stiffness of 2 stents reduces kinking and luminal compression, and the potential space between the stents likely preserves flow around as well as through them. MATERIALS AND METHODS: Four patients recently presented with ureteral obstruction secondary to nonurinary tract malignancies. Previous stenting with a single 6F Double-J* stent had failed in all cases. Three patients experienced flank pain and 1 had persistent azotemia within 3 days of initial stent placement. All patients had significant residual sonographic hydronephrosis despite good stent position. In all cases cystoscopy/stent exchange was performed under local anesthesia with intravenous sedation. Parallel 4.7F Double-J stents were placed simultaneously over 2, 0.035 hydrophilic coated glide wires under fluoroscopic guidance after removal of the malfunctioning 6F stent. RESULTS: Stent placement was uneventful in all 4 patients with prompt drainage of contrast material seen after parallel ipsilateral stent placement. Patients tolerated the double 4.7F parallel stents with no discernible difference in irritative symptoms compared to single 6F stents. Flank pain and azotemia resolved in 3 patients, and hydronephrosis improved in all 4 after placement of parallel Double-J stents. All patients remain alive with a mean followup of 5.8 months (range 4 to 8). Except for 1 patient who later underwent ureterolysis, each has subsequently had the stent changed every 3 months. No patient has required proximal urinary diversion (that is percutaneous nephrostomy tube). CONCLUSIONS: Placement of 2 ipsilateral parallel ureteral stents simultaneously is an easy technique. It may obviate percutaneous nephrostomy tube placement in patients in whom drainage with a single stent failed, especially in cases of extrinsic ureteral compression. PMID- 9400467 TI - Inguinal scrotal incision for penile fracture. AB - PURPOSE: We report a new incision for repair of penile fracture. MATERIALS AND METHODS: We describe 2 cases in which the inguinal scrotal incision was used for repair of penile fracture. The preoperative evaluation as well as the technical case and rationale for use of this incision are discussed. RESULTS: Preoperative cavernosogram delineated the site of the fracture. Immediate repair of the fracture using the inguinal scrotal incision was successful. CONCLUSIONS: The inguinal scrotal incision should be entertained for cases of penile fracture. It avoids incision into markedly edematous penile skin and allows for excellent visualization of the fracture site. PMID- 9400468 TI - Primary skin closure of a large groin defect after inguinal lymphadenectomy for penile cancer using a skin stretching device. AB - PURPOSE: We describe a method of primary closure of a large skin defect. MATERIALS AND METHODS: A 44-year-old man was treated for penile cancer. After left inguinal lymphadenectomy a large skin defect in the groin remained that could not be closed primarily. Allowing skin to stretch beyond its inherent extensibility with a skin stretching system (Sure-Closure) the wound edges were closed without tension. RESULTS: Primary closure of a large skin defect was possible after cyclic stretching and relaxation of the skin with a skin stretching device. CONCLUSIONS: Large skin defects can be closed primarily using a skin stretching device. The primary advantage of the procedure are its simplicity and avoidance of the need for more complicated reconstructive surgery. PMID- 9400469 TI - Microsurgical vasovasostomy: the microdot technique of precision suture placement. AB - PURPOSE: A technique of vasovasostomy that facilitates precision suture placement is presented. MATERIALS AND METHODS: The technique involves mapping of the planned suture exit points with "microdots" placed on the cut ends of the vas deferens with a microtip marking pen. Microdots are placed at 12, 3, 6 and 9 o'clock positions. Four additional dots are placed between each of the previous 4 dots. Exactly 8 mucosal sutures (double armed 10-zero monofilament sutures) are used for each anastomosis. The anastomosis is completed with 8 muscularis sutures (9-zero monofilament) and 6 to 8 sutures (6-zero monofilament) approximating the vasal sheath. RESULTS: In a series of 194 consecutive vasovasostomy procedures using this technique a patency rate of 99.5% was achieved. Pregnancy rates of 54% (crude) and 64% (excluding female factor infertility) were observed for the first 100 subjects of this cohort. CONCLUSIONS: The microdot technique ensures precision suture placement and facilitates the anastomosis of lumens of discrepant diameters by exact mapping of each planned suture. The microdot method separates the planning from the placement. Patency rates using the microdot technique approach 100%. PMID- 9400470 TI - Morbidity of the evaluation of the lower urinary tract with transurethral multichannel pressure-flow studies. AB - PURPOSE: The aim of this prospective study was to determine morbidity and complication rate of invasive urodynamic evaluation of the lower urinary tract after transurethral multichannel pressure-flow studies. MATERIALS AND METHODS: The study included 63 men with the clinical diagnosis of benign prostatic hyperplasia and 56 women with stress urinary incontinence. All patients underwent routine pressure-flow study as part of the urodynamic evaluation. A week later the patients returned for followup which also included a detailed interview on post-evaluation morbidity. RESULTS: The overall complication rate, including urinary retention, gross hematuria, urinary tract infection and fever, was 19.0% (12 of 63) for men and 1.8% (1 of 56) for women. In men there was no statistically significant correlation between post-void residual urine or age and complication rate (p > 0.05). Of the men 4.8% experienced post-investigational urinary retention and all of them had significant bladder outflow obstruction. In addition, obstructed men reported a higher incidence of dysuria and pain (76.2%, 32 of 42) compared to those without obstruction (57.1%, 12 of 21), whereas only 53.6% of women reported these complaints. Of the 63 men 4 (6.2%) had significant urinary tract infections, while only 1 woman (1.8%) had infections. CONCLUSIONS: Invasive urodynamic investigation is associated with a considerable rate of complications and morbidity, particularly in men with infravesical obstruction. These facts must be considered and discussed with the patient before urodynamic testing. PMID- 9400471 TI - Renal autotransplantation and pyelocystostomy for the treatment of urothelial tumors of the upper urinary tract. PMID- 9400472 TI - Hypogastric arterial-ureteral fistula. PMID- 9400473 TI - Laparoscopic urinary undiversion. PMID- 9400474 TI - Massive bleeding from ileal conduit peristomal varices: successful treatment with the transjugular intrahepatic portosystemic shunt. PMID- 9400475 TI - Gastric neobladder for treatment of tuberculosis cystitis. PMID- 9400476 TI - Extensive squamous cell carcinoma in situ of the bladder masking deeply invasive disease. PMID- 9400477 TI - Transitional cell carcinoma of the bladder with involvement of vasa deferentia. PMID- 9400478 TI - Simultaneous bilateral testicular torsion in an adult. PMID- 9400479 TI - Re: Epithelial inclusion cyst formation after free vaginal wall swing sling procedure for stress urinary incontinence. PMID- 9400480 TI - Re: The resistance index represents the corporeal pressure and not the cavernous wall resistance. PMID- 9400481 TI - Re: Possible mechanisms of action of transurethral needle ablation of the prostate on benign prostatic hyperplasia symptoms: a neurohistochemical study. PMID- 9400482 TI - Re: The safety of overnight hospitalization for transurethral prostatectomy: a prospective study of 200 patients. PMID- 9400483 TI - Constant elevation in renal pelvic pressure induces an increase in urinary N acetyl-beta-D-glucosaminidase in a nonobstructive porcine model. AB - PURPOSE: To clarify the physiological significance of renal pelvic pressure elevations encountered in the evaluation of hydronephrotic kidney we examined the effects of different levels of renal pelvic pressure on the induction of renal injury. MATERIALS AND METHODS: A nonobstructive porcine model was created in which the urine drained against a constant predetermined pressure gradient. Renal pelvic pressure of 10, 20 and 40 cm. was created in 2, 2 and 4 animals, respectively. During 18 to 23 hours serial urinary N-acetyl-beta-D glucosaminidase levels were determined as an indicator of renal tubular injury. Tissue specimens were examined histologically and renal arterial blood flow was monitored. RESULTS: Urinary N-acetyl-beta-D-glucosaminidase levels in the kidneys subjected to 10 cm. water remained essentially unchanged. However, at 20 and 40 cm. water statistically significant increases were observed. Similarly, renal arterial blood flow was unchanged at 10 cm. water but it became significantly lower than in controls at 20 and 40 cm. water. Histological evaluation revealed mild to moderate tubular dilatation in the kidneys subjected to 20 and 40 cm. water. CONCLUSIONS: Excessively high collecting system pressure induced renal cellular injury, as reflected by an increase in urinary N-acetyl-beta-D glucosaminidase levels. While renal pelvic pressure up to 10 cm. water appeared to be innocuous, renal cellular injury was evident within as little as 1 hour at renal pelvic pressures 20 cm. water or greater. The degree of N-acetyl-beta-D glucosaminidase in the urine also correlated with a decrease in renal arterial blood flow. PMID- 9400484 TI - Diagnosing the combination of renal dysgenesis, Gartner's duct cyst and ipsilateral mullerian duct obstruction. AB - PURPOSE: We describe the differential points in the diagnosis of the combination of renal dysgenesis, Gartner's duct cyst and ipsilateral mullerian duct obstruction. Various imaging studies and urological procedures were performed. We report our experience in detecting these anomalies in 10 girls and review the literature. MATERIALS AND METHODS: Ten girls, 7 to 13 years old, with this combination of anomalies were identified in the last 10 years. Imaging studies as well as urological procedures were selectively performed, especially at puberty following menarche. Patients received long-term followup with ultrasound. RESULTS: Cystic dilation of Gartner's duct protruded into the bladder and presented as a ureterocele in 5 patients and posterior to the bladder in 5. Surgical removal of a partial portion of a Gartner's duct cyst was performed in 5 patients for alleviation of urinary symptoms. Unilateral mullerian duct obstruction was demonstrated in all 10 patients. Excision of the vaginal septum was performed in 6 patients for relief of genital obstruction. CONCLUSIONS: When cystic dilatation of the pelvis, especially a ureterocele-like cyst without ureteral dilatation, is found in girls with ipsilateral renal dysgenesis, the possibility of a Gartner's duct cyst should be considered. For early detection and treatment of unilateral obstruction of duplicated mullerian ducts pelvic sonography should be performed at puberty, especially just after menarche, in girls with renal dysgenesis and ipsilateral Gartner's duct cyst. PMID- 9400486 TI - Retroperitoneal fibrosis mimicking recurrent leukemia in a 7-year-old boy. PMID- 9400485 TI - Surgery versus observation for managing obstructive grade 3 to 4 unilateral hydronephrosis: a report from the Society for Fetal Urology. AB - PURPOSE: The Society for Fetal Urology has undertaken the first multicenter prospective randomized study of high grade obstructive unilateral hydronephrosis to evaluate the natural history of untreated obstruction and compare it to the benefits of pyeloplasty. MATERIALS AND METHODS: Since 1991, infants with isolated unilateral Society for Fetal Urology grade 3 hydronephrosis and ipsilateral obstruction with greater than 40% differential renal function on well tempered renography were studied. Patients were randomly assigned to observation or pyeloplasty groups. Renal ultrasound and well tempered renography were performed biannually for 1 year and yearly thereafter. Crossover criteria for surgery included concurrent worsening of isotope washout and increasing grade of hydronephrosis or a greater than 10% point loss in percent differential renal function that was noted between studies. The end point of the study was the 3 year anniversary of randomization. RESULTS: A total of 32 infants from 10 centers were randomized equally to 2 groups. The starting grade of hydronephrosis and percent differential renal function were similar between the 2 groups. At 6 months and 1 year the grade of hydronephrosis was significantly reduced (p < 0.02) and well tempered renography was significantly more likely to demonstrate no obstruction (p < 0.03) in the surgical group compared with the observation group. The mean percent differential renal function remained stable and similar in both groups. Reduced hydronephrosis and resolution of obstruction in the surgery group persisted as a trend at the 2 and 3-year anniversaries. In the observation group 4 patients (25%) showed enough renal deterioration to qualify for crossover to surgery. CONCLUSIONS: Infant pyeloplasty significantly improved the grade of hydronephrosis and drainage pattern at 6 months and 1 year postoperatively, when compared with observation. Renal function stabilization was similar for either management approach. However, 25% of the patients satisfied objective criteria of status deterioration requiring pyeloplasty. PMID- 9400487 TI - Subclinical changes in bladder function in children presenting with nonurological symptoms of the tethered cord syndrome. AB - PURPOSE: We evaluated the urodynamic findings in myelodysplastic children with the tethered cord syndrome without urological symptoms to determine if occult bladder changes occur or if routine preoperative urodynamic evaluation is not indicated for this select population. MATERIALS AND METHODS: Preoperative and postoperative urodynamic studies were performed on children with myelodysplasia and the tethered cord syndrome between 1988 and 1994. Inclusion criteria were neurological or musculoskeletal surgical indications only, without urological status changes, radiographic confirmation of the tethered cord syndrome, and water cystometry performed preoperatively within 1 week and again postoperatively within 6 months. The parameters of interest included total bladder capacity and pressure, leak point pressure, compliance, uninhibited contractions, electromyelogram activity and sensation. RESULTS: A total of 20 children, 11 girls and 9 boys, 2.3 to 17.3 years old were included in the study. Worsening scoliosis and lower extremity weakness were the most common presentations. Urodynamic studies were conducted 1.8 days preoperatively (mean) and 104.3 days postoperatively (mean). Results were analyzed with regard to improvement or deterioration between preoperative and postoperative urodynamic studies. Of the 20 children 15 (75%) demonstrated improvement between the 2 urodynamic studies, including 10 who improved in 1 parameter (most often with resolution of uninhibited contractions), 3 in 2, 1 in 3 and 1 in 4. There were no significant postoperative changes for any of the specific parameters. Urodynamic studies identified 7 children with preoperative leak point pressures above 40 cm. water, of whom only 2 had decreased pressures below 40 cm. water, 2 had postoperative deterioration of compliance and 1 had preoperative detrusor-sphincter dyssynergia. CONCLUSIONS: Routine preoperative and postoperative urodynamic evaluations in children with the tethered cord syndrome without clinical changes to urological status may be important. The majority of clinically asymptomatic children will demonstrate preoperative urodynamic findings that improve postoperatively, which serves as another marker of progress after spinal cord untethering. Moreover, some asymptomatic children will demonstrate changes to the urinary tract that merit management changes, such as detrusor-sphincter dyssynergia, elevated bladder storage pressures and poor compliance, which may have otherwise been delayed in recognition. PMID- 9400488 TI - Fluorescence in situ hybridization on nuclei from paraffin-embedded tissue in low stage pure embryonal carcinoma of the testis. AB - Approximately 30% of patients who present with clinical stage A nonseminomatous testis cancer are in fact pathologic stage B. In previous studies an increasing volume of embryonal carcinoma in the orchiectomy specimen was associated with a higher likelihood of being pathologic stage B. However, not all patients with pure embryonal carcinoma in the primary tumor were pathologic stage B. In an effort to discriminate patients with pure embryonal carcinoma in the testicular specimen relative to pathologic stage, archival specimens from patients presenting with clinical stage A pure embryonal carcinoma were examined by fluorescence in situ hybridization (FISH) with newly developed probes for chromosome arms 12p and 12q. Whole nuclei from archival material from 14 patients (six pathologic stage A, seven pathologic stage B and one stage C) with 100% embryonal carcinoma in the orchiectomy specimen were studied using bicolor FISH with chromosome arm 12p- and 12q-specific painting probes developed by chromosome microdissection. In all cases a blinded analysis showed distinct regions of 12p and 12q probe hybridization simultaneously and allowed identification of probable normal chromosomes 12, as well as regions of amplification of 12p sequences, including possible i(12p). In 5/14 specimens, a distinct and peculiar pattern of 12p hybridization was observed which resembled 12p "disarray" or "multifocal 12p". Of the five specimens demonstrating multifocal 12p, four were pathologic stage B, while one was pathologic stage A. Whether the trend toward multifocal 12p predicts metastatic potential in primary testicular embryonal carcinoma will need to be assessed using a larger series of patients. PMID- 9400489 TI - Low frequency of positive telomerase activity in a chromophobe subtype of renal cell carcinoma. AB - PURPOSE: In malignant tumors, telomerase reactivation plays an important role in the acquisition of cellular immortality. We evaluated the telomerase activity in renal cell carcinomas (RCCs) with special reference to their clinicopathologic features. MATERIALS AND METHODS: Telomerase activity was examined in 47 RCCs and 9 RCC cell lines by telomeric repeat amplification protocol assay (TRAP). The telomere lengths were assessed by Southern analysis of terminal restriction fragments (TRFs) generated by Hinfl-digested DNA. RESULTS: Thirty-six (77%) of the 47 RCCs and all 9 RCC cell lines showed telomerase activity, whereas no activity was detected in any of 30 normal kidneys. When the tumors were histopathologically classified, only one (17%) of the 6 chromophobe cell carcinomas was telomerase-positive. This frequency was significantly low (p < 0.001) when compared with those in clear cell RCCs (93%; 26/28). In 40 of the 47 patients, DNA from the tumor tissues and the paired normal kidneys was available for analysis of the TRF lengths. No tumor showed elongated TRF length compared to its paired normal kidney. Regarding the relationship between telomere length and telomerase activity, 23 (74%) of the 31 telomerase-positive RCC and 6 (67%) of the 9 telomerase-negative RCC exhibited reduced TRF. There was no significant correlation between the telomere reduction and telomerase activity. CONCLUSION: The mechanism for preventing telomere shortening may differ according to RCC subtype. Alternatively, telomerase-negative tumors may have yet to reach the immortal stage when they progress to clinical cancer. The telomerase activity status may contribute to the biological potential and the prognosis of RCCs. PMID- 9400490 TI - Bethanechol activates a post-receptor negative feedback mechanism in rabbit urinary bladder smooth muscle. AB - PURPOSE: Recent studies using vascular and gut smooth muscles indicate that contractile receptor agonists may activate post-receptor down-regulatory mechanisms causing a temporary reduction in the strength of subsequent contractions. Our data indicate a similar mechanism exists in detrusor smooth muscle of the urinary bladder. MATERIALS AND METHODS: Each isolated strip of female rabbit detrusor was placed in a tissue bath, secured to an isometric force transducer, and length-adjusted until depolarization with 110 mM KCl produced a maximum contraction (S0). Subsequent contractions were normalized to S0 (S/S0) or to a first stimulus with 30 mM KCl or caffeine (S/S1). Tissues were pretreated with the muscarinic receptor agonist, bethanechol (BE), then stimulated with KCl, caffeine, or Bay k 8644 to identify potential post-receptor down-regulation. RESULTS: Contractions induced by 30 mM KCl had three phases labeled fast peak (FP), slow peak (SP) and steady-state (SS). In tissues exposed for 30 min. to a maximum BE concentration then washed for 5 min., the KCl-induced FP and SP, but not SS, responses were reduced by approximately 40%. Smaller reductions in peak KCl-induced contractions occurred in tissues pretreated for a shorter duration or with a 100-fold lower BE concentration. This down-regulation induced by bethanechol pretreatment was reversible, lasting approximately 1-2 h. Not only were KCl-induced contractions reduced by BE pretreatment, but also those produced by the intracellular Ca(2+)-mobilizer, caffeine, and the L-type Ca2+ channel agonist, Bay k 8644. CONCLUSIONS: Pretreatment of isolated strips of rabbit detrusor with a muscarinic receptor agonist produced short-term down-regulation of KCl-induced peak contractions that may have involved inhibition of both influx of extracellular Ca2+ and release of intracellular Ca2+. Reductions in the degree of this novel modulatory response during disease conditions and aging could enhance contractile activity, possibly causing detrusor instability. PMID- 9400491 TI - Detailed interstitial temperature mapping during treatment with a novel transurethral microwave thermoablation system in patients with benign prostatic hyperplasia. AB - PURPOSE: To delineate in detail the temperature changes in the prostate gland and adjacent structures during treatment with a newly designed microwave thermoablation system in patients with benign prostatic hyperplasia (BPH). MATERIALS AND METHODS: Microwave thermoablation treatment was administered to 22 BPH patients at two centers in the U.S. and Argentina using the Urologix Targis targeted transurethral thermoablation system. Continuous temperature measurements were made with widely spatially dispersed fiber optic thermosensors at 11 to 24 prostatic sites in each patient using a recently described accurate stereotactic method. Urethral and rectal temperatures were also measured. RESULTS: Treatment using the microwave thermoablation system resulted in marked elevation of intraprostatic temperatures to as high as 80C in some patients with little or no elevation of urethral or rectal temperatures. Average temperature increased with radial distance from the urethra to a peak at 5 to 7 mm. and declined exponentially at greater distances. Higher maximum intraprostatic temperatures in individual patients were associated with a larger zone, up to 24.0 mm. in radius, of prostatic tissue exposed to thermoablative temperatures of 45C and higher. Along the longitudinal axis of the microwave treatment catheter, thermoablative temperatures were confined to a zone of 11.5 mm. from the microwave antenna midpoint apically and 11.3 mm. basally, that is, a range shorter than the length of the treatment catheter's microwave antenna (2.8 to 3.5 cm.). The mean temperature in the posterior sector of the prostate gland during treatment (43.6C; 95% CI, 41.1 to 46.1C) was significantly lower (p < 0.05) by 6.7C than that in the anterolateral prostate (50.3C; 95% CI, 48.3 to 52.3C), as a consequence of the preferential heating design of the treatment catheter. Intraprostatic mean temperature during treatment, as measured at all thermosensor sites without respect to spatial location, was 47.1C (95% CI, 44.2 to 50.0C), a value significantly higher (p < 0.05) than that measured in the urethra (39.6C; 95% CI, 36.6 to 42.6C) or rectum (37.7C; 95% CI, 36.7 to 38.7C). There was a strong correlation between the temporal pattern of fluctuation in urethral temperature and that of prostate temperature (r = 0.83; p < 0.001) during treatment. CONCLUSIONS: Treatment with the microwave thermoablation system fulfilled the requirements for an effective and safe microwave-based BPH treatment modality by exposing obstructive tissue to high temperatures without endangering vulnerable adjacent tissues. PMID- 9400492 TI - Does topical instillation therapy influence chromosomal aberrations in superficial bladder cancer? AB - PURPOSE: Patients with a high risk for superficial bladder cancer are treated by topical immuno-or chemotherapy after transurethral resection to reduce the chance of recurrence and/or progression. The aim of this study was to analyse if cytogenetical abnormalities, which are known to be constantly related to bladder cancer, are modified or eliminated by topical immuno- or chemotherapy. MATERIALS AND METHODS: Using fluorescence in situ hybridization (FISH), the influence of topical instillation therapy with Bacillus Calmette-Guerin (BCG) and Mitomycin C (MMC) on numerical aberrations of chromosomes 7, 9 and 17 was investigated in 25 patients with transitional cell cancer (TCC) of the bladder. Data were compared with histological and clinical outcome. Fifteen TCC patients with similar histological criteria without instillation therapy served as controls. Median follow-up was 30 +/- 2 months. RESULTS: After BCG treatment 10 of 15 patients (66.6%) developed recurrent and 2/15 (13.3%) progressive disease. Three of 15 patients (20.0%) had no evidence of disease. Numerical aberrations did not change in 8 of the 15 BCG patients (53.3%) and changed to a more aggressive pattern in 40.0% (6/15). Five of 10 MMC treated patients (50.0%) developed a recurrent tumor, 2/10 (20.0%) progressed and 3/10 (30.0%) had no evidence of disease. Four of 10 (40.0%) of these patients showed stable and 5/10 (50.0%) progressive chromosomal patterns. Only one patient in each group with primary chromosomal alterations changed to a regular diploid chromosomal pattern after therapy according to a complete clinical remission. CONCLUSION: Even though topical immuno- and chemotherapy may be useful to delay recurrence and progression, chromosomal patterns remain basically unstable. PMID- 9400493 TI - Enhanced expression of the mesenchymal marker, vimentin, in hyperplastic versus normal human prostatic epithelium. AB - A total of 20 benign prostatic hyperplasia (BPH) tissue samples from 13 patients were evaluated for the coexpression of cytokeratins and vimentin intermediate filaments in the glandular epithelium. Vimentin expression was significantly increased (p < 0.001) in BPH epithelium compared to adjacent "normal" epithelium. The data imply that epithelial/mesenchymal (E/M) transformations may play a role in the pathogenesis of BPH. PMID- 9400494 TI - The effects of brefeldin A (BFA) on cell cycle progression involving the modulation of the retinoblastoma protein (pRB) in PC-3 prostate cancer cells. AB - PURPOSE: To investigate the effects of brefelding A (BFA) on the growth of the androgen-independent human prostate cancer PC-3 cells, focusing on cell cycle regulation. MATERIALS AND METHODS: BFA is a fungal macrocyclic lactone with an antiviral activity. PC-3 cells were cultured with various concentrations of BFA for indicated times and cell growth was monitored at each time point. Cell cycle analysis was performed to explore the mechanism of BFA-induced growth inhibition. To further investigate the cell cycle regulation, cell cycle-controlling factors, such as the retinoblastoma gene product (pRB) and its regulatory components cdk2, cdk4, and cyclin D1, were analyzed by Western immunoblots. RESULTS: BFA was a potent growth inhibitor at a concentration of 30 ng./ml., resulting in a > 70% reduction in cell number at 3 days. Cell cycle analysis revealed a cell arrest in the G1 to S phase transition. Western blots further showed that BFA induced dephosphorylation of pRB accompanied by down regulation of cdk2, cdk4, and cyclin D1 expression. The extended pRB dephosphorylation in control cell lysates was also observed by the addition of BFA-treated lysates, but was prevented by the inclusion of phosphatase inhibitors in assay mixtures. CONCLUSION: These results suggest that BFA may be a potent cell cycle modulator, which post-translationally regulates pRB phosphorylation possibly by down-regulating cdk2, cdk4, and cyclin D1 and/or by up-regulating a phosphatase(s) capable of dephosphorylating pRB. Thus, BFA-induced growth inhibition in PC-3 cells appears to be at least partially due to the modulation of a pRB-mediated growth pathway. PMID- 9400495 TI - Polymerase chain reaction amplification of bacterial 16S rRNA genes in interstitial cystitis and control patient bladder biopsies. AB - PURPOSE: Several characteristics of the chronic bladder disease called interstitial cystitis (IC) suggest an infectious etiology. However, a single causative organism has not been convincingly cultured in vitro, and DNA for a variety of microorganisms has been found inconsistently in bladder biopsies from IC patients. We therefore looked for a possible bacterial cause for IC by using a sensitive nested PCR assay on cystoscopic bladder biopsy specimens obtained from IC patients and controls. MATERIALS AND METHODS: Bladder biopsies were obtained at cystoscopy from 6 IC patients and 6 controls. DNA was extracted from these specimens and PCR with 2-round amplification performed using nested primers from a highly conserved region of the bacterial 16s rRNA gene. Amplified DNA was purified and sequenced using the Sequenase PCR Product Sequencing Kit, and the sequences obtained were compared with bacterial rRNA gene sequences recorded in GenBank. RESULTS: Biopsy specimens from all 6 patients and 6 controls were positive by PCR for DNA encoding bacterial 16s rRNA. Sequence data indicated a predominant microorganism in 10 of the 12 specimens, with > 95% homology to DNA from several different genera of bacteria including Acinetobacter, Propionobacterium, Salmonella, and Escherichia. None of the organisms identified by PCR had been cultured from tissue or urine obtained simultaneously from these persons, using sensitive culture techniques. CONCLUSIONS: These data indicate no difference between IC patients and controls in the proportion of bladder biopsies with PCR positivity or the type(s) of organism present, providing additional evidence that IC is not associated with infection by a particular type of bacterium. PMID- 9400496 TI - Autologous transplantation of urothelium into demucosalized gastrointestinal segments: evidence for epithelialization and differentiation of in vitro expanded and transplanted urothelial cells. AB - PURPOSE: Our study established a technique for in vitro expansion and subsequent transplantation of autologous urothelial cells into vascularized seromuscular segments from stomach and colon in sheep. The proof of proliferation and differentiation of the transplanted urothelium in the absence of resident urothelium is considered to be a prerequisite for use of this technique in bladder augmentation. MATERIALS AND METHODS: Autologous sheep urothelial cells were expanded in vitro and grown on collagen membranes for sheet grafting. Using a vital stain, viability and confluency status of the urothelial graft were determined before transplantation into demucosalized segments isolated from the sheep stomach and colon gastrointestinal pouches. The gastrointestinal segments were sewn up and remained in the abdomen as small pouches stiched to the abdominal wall. Take and differentiation of transplanted cells within the pouch were assessed two and three weeks later using histological and immunohistological means. RESULTS: Urothelial cells grew well on collagen membranes. A confluency status > 40% and co-culturing with 3T3 feeder cells favored successful transplantation. Two weeks after transplantation a multilayered urothelial-like epithelium was found to line the lumen of the pouch. The epithelium was characterized by a distinct urothelium-typical distribution of basal and luminal keratins and the expression of the umbrella cell-specific marker uroplakin III. Moreover, the epithelium had an underlying basal lamina which focally contained collagen type IV. CONCLUSIONS: The data indicate that in vitro expanded urothelial cells are capable of epithelializing demucosalized gastrointestinal segments forming a genuine, differentiated "neo" urothelium. PMID- 9400497 TI - Expression of transforming growth factor alpha and epidermal growth factor receptor in adult polycystic kidney disease. AB - Adult polycystic kidney disease (APKD) is a common genetic disease with a frequency of 1:1000. Evidence suggests that transforming growth factor alpha (TGF alpha) signaling may contribute to the hyperproliferation of the cystic epithelia in APKD. TGF alpha and epidermal growth factor (EGF) are well known mitogens expressed in the kidney and both exert their biological activities through binding to the same EGF receptor. A transgenic mouse that over-expressed TGF alpha developed renal cysts; raised levels of TGF alpha and EGF receptor mRNA were found in kidneys from two autosomal dominant APKD patients. To study the role of TGF alpha in cyst formation, we analyzed nine anatomically diagnosed adult polycystic kidneys and four normal kidneys using immunohistochemistry. We also traced the possible origins of the cysts by staining with the proximal convoluted tubule (PCT) marker, gp330, and the distal convoluted tubule (DCT) and collecting tubule (CT) marker, peanut agglutinin (PNA). In normal kidneys, TGF alpha protein was concentrated in the DCT and CT and EGF receptor protein in all three tubule types. In the early cysts of APKD, the cystic epithelia showed strong positive staining with TGF alpha, EGF receptor and gp330 but negative with PNA. Strong TGF alpha and EGF receptor staining was also found in the mixture of advanced cysts in the end-stage cystic kidneys although the cysts are likely to be derived from different segment of the renal tubules. This increased TGF alpha and EGF receptor expression in all cases and all types of cysts suggests that autocrine/paracrine stimulation by TGF alpha may be a common mechanism in cyst development in APKD. PMID- 9400498 TI - Prostate-specific antigen forms complexes with human alpha 2-macroglobulin and binds to the alpha 2-macroglobulin receptor/LDL receptor-related protein. AB - PURPOSE: To investigate the binding of the prostate-specific antigen (PSA) to human alpha 2-macroglobulin (alpha 2-M) and to alpha 1-antichymotrypsin (ACT). MATERIALS AND METHODS: Binding analysis was evaluated by electrophoresis, Western blotting, enzyme-linked immunosorption assay (ELISA) and size exclusion chromatography. Quantification of PSA and of different forms of alpha 2-M was performed using commercial test kits. The cleavage site of PSA in alpha 2-M was analyzed by SDS-PAGE and microsequencing. RESULTS: Binding of PSA to alpha 2-M is initiated by the cleavage of the peptide bond between amino acids Tyr 686 and Glu 687 of the bait region indicating a chymotrypsin-like activity of the PSA. The PSA's proteolytic cleavage triggers the transformation of alpha 2-M as detected by conformation-specific monoclonal antibodies. Kinetic analysis revealed faster binding of PSA to alpha 2-M than to ACT. The PSA bound to alpha 2-M is caged by the inhibitor and thus escapes detection by antibodies. This results in an incorrect calculation of the level of PSA when released from prostate into the blood. Complexes of PSA-alpha 2-M and PSA-ACT were found to bind to the alpha 2 macroglobulin receptor/LDL receptor-related protein (alpha 2-M-R/LRP) which may be the clearance receptor for PSA. CONCLUSIONS: Quantifying free PSA and PSA-ACT complexes, as routinely done in managing prostate-associated diseases, does not represent the total secretion capacity of the prostate. The proteinase inhibitor alpha 2-M has to be considered as a main contributor to PSA complex formation in the blood. PMID- 9400499 TI - Patient-reported impotence and incontinence after nerve-sparing radical prostatectomy. PMID- 9400500 TI - Endoluminal sonographic evaluation of ureteral and renal pelvic neoplasms. PMID- 9400501 TI - Mysterious, unseen, and quite possibly unpleasant. PMID- 9400502 TI - Recommendations on folate intake. PMID- 9400503 TI - Counselling in primary care. PMID- 9400504 TI - Vancomycin-resistant Staphylococcus aureus: apocalypse now? PMID- 9400506 TI - Half-way at Kyoto--but to where? PMID- 9400505 TI - European perspectives on general medicine. PMID- 9400507 TI - Communicating drug-safety information. PMID- 9400508 TI - Adjuvant chemotherapy for localised resectable soft-tissue sarcoma of adults: meta-analysis of individual data. Sarcoma Meta-analysis Collaboration. AB - BACKGROUND: Individually, randomised trials have not shown conclusively whether adjuvant chemotherapy benefits adult patients with localised resectable soft tissue sarcoma. METHODS: A quantitative meta-analysis of updated data from individual patients from all available randomised trials was carried out to assess whether adjuvant chemotherapy improves overall survival, recurrence-free survival, and local and distant recurrence-free intervals (RFI) and whether chemotherapy is differentially effective in patients defined by age, sex, disease status at randomisation, disease site, histology, grade, tumour size, extent of resection, and use of radiotherapy. FINDINGS: 1568 patients from 14 trials of doxorubicin-based adjuvant chemotherapy were included (median follow-up 9.4 years). Hazard ratios of 0.73 (95% CI 0.56-0.94, p = 0.016) for local RFI, 0.70 (0.57-0.85, p = 0.0003) for distant RFI, and 0.75 (0.64-0.87, p = 0.0001) for overall recurrence-free survival, correspond to absolute benefits from adjuvant chemotherapy of 6% (95% CI 1-10), 10% (5-15), and 10% (5-15), respectively, at 10 years. For overall survival the hazard ratio of 0.89 (0.76-1.03) was not significant (p = 0.12), but represents an absolute benefit of 4% (1-9) at 10 years. These results were not affected by prespecified changes in the groups of patients analysed. There was no consistent evidence that the relative effect of adjuvant chemotherapy differed for any subgroup of patients for any endpoint. However, the best evidence of an effect of adjuvant chemotherapy for survival was seen in patients with sarcomas of the extremities. INTERPRETATION: The meta analysis provides evidence that adjuvant doxorubicin-based chemotherapy significantly improves the time to local and distant recurrence and overall recurrence-free survival. There is a trend towards improved overall survival. PMID- 9400509 TI - Human herpesvirus 8 variants in sarcoid tissues. AB - BACKGROUND: The cause of sarcoidosis is unknown, although mycobacteria have been implicated. We examined sarcoid tissues for human herpesvirus 8 (HHV-8) in addition to mycobacterial genomic sequences. METHODS: Biopsy samples from 17 patients with sarcoidosis were studied (eight transbronchial, 27 lymph node, two skin, and two oral mucosa). We used tissues (n = 137) from 96 patients without sarcoidosis as negative controls. A nested PCR was applied to amplify a segment of open reading frame (ORF) 26 of the HHV-8 genome, and a heminested PCR was to amplify a segment of ORF 25 of HHV-8 and of the 16 S rRNA gene of mycobacteria. Differences in base sequences of the amplified fragments were resolved with single-strand conformation polymorphism and dideoxy sequencing. FINDINGS: HHV-8 ORF 26 DNA was detected in significantly higher proportions of sarcoid than of non-sarcoid tissue samples from lung (8/8 vs 0/54; p < 0.0001), lymph nodes (26/27 vs 6/29; p < 0.0001), skin (2/2 vs 0/17; p = 0.006), and oral tissues (2/2 vs 1/13; p = 0.029). 31 (82%) of the 38 ORF 26 DNA-positive sarcoid specimens were also positive for ORF 25 DNA. For mycobacteria-like 16 S rRNA DNA, the proportion positive was significantly higher in sarcoid than non-sarcoid tissues for lymph node samples (11/27 vs 2/29; p = 0.003) but not for other tissues (lung 3/8 vs 22/54; skin 2/2 vs 15/17; and oral tissues 1/2 vs 0/13). Overall, the prevalence of HHV-8 ORF 26 sequences was higher in sarcoid tissues than in non sarcoid tissues (p < 0.0001). When patients whose tissues were included in a masked phase of the study were treated as units of analysis, eight of eight sarcoidosis patients were positive for HHV-8 ORF 26 DNA, compared with three of 56 control patients (p < 0.0001); for mycobacteria-like sequences, three of eight sarcoidosis patients were positive, compared with four of 56 controls (p = 0.0464). The HHV-8 ORF 26 sequences, ten of which were unique, could be segregated into four groups according to peptide motifs. In seven of nine patients from whom biopsy samples were taken from various sites, different sequences were recovered. The mycobacterial sequences amplified from sarcoid tissues were also varied, but none was homologous to those of known species. INTERPRETATION: Variant HHV-8 DNA sequences are found in a wide range of sarcoid but not non-sarcoid tissues. Mycobacteria-like 16 S rRNA sequences are more frequently present in sarcoid lymph nodes and not in other tissue types, but do not indicate infection by a particular mycobacterial species. PMID- 9400510 TI - Randomised controlled assessment of non-directive psychotherapy versus routine general-practitioner care. AB - BACKGROUND: We compared the efficacy of and patients' satisfaction with general practice-based psychotherapists with those of general practitioners in providing treatment to people with emotional difficulties. METHODS: We carried out a prospective, randomised, controlled trial of brief, non-directive psychotherapy and routine general-practice care. Therapists adhered to a non-directive Rogerian model of psychotherapy. Between one and 12 sessions of psychotherapy were given over 12 weeks in 14 general practices in north London, UK. Of 136 patients with emotional difficulties, mainly depression, 70 patients were randomly assigned to the therapist and 66 to the general practitioner. Depression, anxiety, other mental-disorder symptoms, and social adjustment were measured by self-report at baseline, 3 months, and 9 months. Patients' satisfaction was also measured by self-report at 3 and 9 months. FINDINGS: All patients improved significantly over time. There were no significant differences between the groups receiving brief psychotherapy and routine general-practitioner care. Patients assigned brief psychotherapy were more satisfied with the help they received than those assigned to the general practitioner at both 3 and 9 months' follow-up (mean scores on satisfaction scale 50.9 [SD 7.9] vs 44.4 [9.8] and 45.6 [9.4] vs 37.1 [11.2], respectively). INTERPRETATION: General-practitioner care is as effective as brief psychotherapy for patients usually referred by doctors to practice-based psychotherapists. Patients with emotional difficulties prefer brief psychotherapy from a counsellor to care from their general practitioner. PMID- 9400511 TI - Minimum effective dose of folic acid for food fortification to prevent neural tube defects. AB - BACKGROUND: Although a daily supplement of 400 micrograms folic acid has been shown to prevent neural-tube defects (NTD), most women do not take the recommended supplement. Thus, food fortification is to be introduced in the USA and is being considered in the UK. Because of safety concerns, the USA has chosen a level of fortification that will increase the average woman's intake by only 100 micrograms. Such an increase, although safe, may be ineffective; but a trial to assess its efficacy would be unethical. Because women with red-cell folate concentrations above 400 micrograms/L have a very low risk of NTD, we undertook a randomised trial of several folic acid doses to find out how much is needed to reach this protective concentration. METHODS: We screened 323 women. 172 with red cell folate between 150 micrograms/L and 400 micrograms/L were invited to take part in the trial. 121 women were randomly assigned placebo or 100 micrograms, 200 micrograms, or 400 micrograms daily of additional folic acid. Compliance was monitored by having the women sign a dated sheet when taking the tablet. 95 women completed the 6-month study. FINDINGS: There were significant increases in red cell folate in all folic acid groups. The placebo group showed no significant change. The median incremental changes and median post-treatment concentrations were 67 micrograms/L (95% CI 43-120) and 375 micrograms/L (354-444) in the 100 micrograms/day group, 130 micrograms/L (108-184) and 475 micrograms/L (432-503) in the 200 micrograms/day group, and 200 micrograms/L (125-312) and 571 micrograms/L (481-654) in the 400 micrograms/day group. INTERPRETATION: A fortification programme that delivered 400 micrograms folic acid daily to women would protect against NTD, but at the expense of unnecessarily high exposure for many people. Delivery of 200 micrograms daily is also effective against NTD and safer for the general population. Based on projections from the positive folate balance in the group that received 100 micrograms daily, this dose taken continually, as it will be in fortified food, will also produce an important decrease in NTD. PMID- 9400512 TI - Dissemination in Japanese hospitals of strains of Staphylococcus aureus heterogeneously resistant to vancomycin. AB - BACKGROUND: Since the discovery of the vancomycin-resistant Staphylococcus aureus (VRSA) strain Mu50 (minimum inhibitory concentration [MIC] 8 mg/L), there has been concern about the potential spread of such strains throughout Japanese hospitals. Two important questions need to be answered: (1) what is the prevalence of VRSA, and (2) by what mechanism does vancomycin resistance occur. METHODS: The vancomycin susceptibilities of three methicillin-resistant S aureus (MRSA) strains (Mu50, Mu3, and H1) and the methicillin-susceptible S aureus type strain FDA209P were compared by MIC determinations and population analysis. Mu3 (MIC 3 mg/L) was isolated from the sputum of a patient with pneumonia after surgery who had failed vancomycin therapy. H1 (MIC 2 mg/L), which is a representative vancomycin-susceptible MRSA strain, was isolated from a patient with pneumonia who responded favourably to vancomycin therapy. Subclones of Mu3 with increased resistance against vancomycin were selected with serial concentrations of vancomycin and their MICs were determined. The prevalence of VRSA and Mu3-like strains in Japanese hospitals was estimated by population analysis from 1149 clinical MRSA isolates obtained from 203 hospitals throughout Japan. The genetic traits of the Mu3 and Mu50 strains were compared with clonotypes of MRSA from around the world. FINDINGS: Mu3 and Mu50 had an identical pulsed-field gel electrophoresis banding pattern. When grown in a drug-free medium, Mu3 produced subpopulation of cells with varying degrees of vancomycin resistance, thus demonstrating natural heterogeneity, or variability, in susceptibility to vancomycin. In the presence of vancomycin, Mu3 produced subclones with resistance roughly proportional to the concentrations of vancomycin used. Selection of Mu3 with 8 mg/L or more of vancomycin gave rise to subclones with vancomycin resistance equal to that of Mu50 (MIC 8 mg/L) at a frequency of 1/1,000,000. During screening of Japanese MRSA strains, no strain of VRSA additional to Mu50 was found. The prevalence of MRSA isolates heterogeneously resistant to vancomycin was 20% in Juntendo University Hospital, 9.3% in the other seven university hospitals, and 1.3% in non-university hospitals or clinics. INTERPRETATION: Heterogeneously resistant VRSA is a preliminary stage that allows development into VRSA upon exposure to vancomycin. Heterogeneously resistant VRSA was found in hospitals throughout Japan. This finding could explain, at least partly, the frequent therapeutic failure of MRSA infection with vancomycin in Japan. PMID- 9400513 TI - Fever, haematuria, proteinuria, and a parrot. PMID- 9400514 TI - Stimulation of subthalamic nucleus alleviates tremor in Parkinson's disease. PMID- 9400515 TI - Chronic stimulation of subthalamic nucleus improves levodopa-induced dyskinesias in Parkinson's disease. PMID- 9400516 TI - Fever associated with intravenous antibiotics in adults with cystic fibrosis. PMID- 9400517 TI - Vocal cord haematoma after thrombolysis. PMID- 9400518 TI - Spherocytosis, splenectomy, strokes, and heat attacks. PMID- 9400519 TI - Sweat electrolyte concentrations in children with atopic dermatitis. PMID- 9400520 TI - Synovial sarcoma of bone delineated by spectral karyotyping. PMID- 9400522 TI - Field trial of a rapid card test for Wuchereria bancrofti. PMID- 9400521 TI - Non-cardiovascular disease mortality and diabetes mellitus. PMID- 9400523 TI - Exhaled nitric oxide during lung transplantation. PMID- 9400525 TI - HIV epidemic could number 40 million by year 2000. PMID- 9400524 TI - Penile rigidity in erectile dysfunction treated with alprostadil. PMID- 9400526 TI - Trial to reduce vertical transmission of HIV-1 on schedule in Uganda. PMID- 9400528 TI - Final Krever report paints picture of regulatory dysfunction. PMID- 9400527 TI - Salt and hypertension: consensus or controversy? PMID- 9400529 TI - Irritable bowel syndrome. PMID- 9400530 TI - Development of a malaria vaccine. AB - Development of an effective malaria vaccine poses a major scientific challenge both in the laboratory and in the field. Such a vaccine is necessary because of the massive disease burden of malaria in the developing world, the global spread of drug resistance, and the difficulty of sustainable control of the mosquito vector. Animal models have shown the immunological feasibility of vaccines targeted against different stages of parasite development, and studies in human volunteers have shown that a recombinant protein vaccine can protect against challenge with the homologous strain of parasite. However, both natural and vaccine-induced immunity are hampered by the remarkable capacity of the parasites to vary critical antigenic structures; large field trials of a synthetic peptide vaccine gave equivocal results. In an attempt to overcome the dual difficulty of poor immunogenicity and parasite diversity, much experimental work is now focused on complex antigenic constructs, delivered as DNA vaccines or in live vectors such as vaccinia, with multiple targets at each stage of parasite development. PMID- 9400531 TI - Modest salt restriction in older people. PMID- 9400532 TI - Modest salt restriction in older people. PMID- 9400533 TI - Modest salt restriction in older people. PMID- 9400534 TI - New-variant Creutzfeldt-Jakob disease and treatment of haemophilia. Executive Committee of the UKHCDO. United Kingdom Haemophilia Centre Directors' Organisation. PMID- 9400535 TI - Hormone replacement therapy and biological aggressiveness of breast cancer. PMID- 9400536 TI - Ventricular remodelling in dilated cardiomyopathy. PMID- 9400537 TI - Ventricular remodelling in dilated cardiomyopathy. PMID- 9400538 TI - Ventricular remodelling in dilated cardiomyopathy. PMID- 9400539 TI - Meningococcal vaccine in sub-Saharan Africa. PMID- 9400540 TI - Meningococcal vaccine in sub-Saharan Africa. PMID- 9400541 TI - Meningococcal vaccine in sub-Saharan Africa. PMID- 9400542 TI - Meningococcal vaccine in sub-Saharan Africa. PMID- 9400543 TI - Sickle-cell disease. PMID- 9400544 TI - Continuing increase in invasive methicillin-resistant infection. PMID- 9400545 TI - End-of-life decisions in Dutch paediatric practice. PMID- 9400546 TI - Sex and examination results. PMID- 9400547 TI - Sex and examination results. PMID- 9400548 TI - Quality of essential drugs. PMID- 9400549 TI - Terathanasia or teratothanasia? PMID- 9400550 TI - The association of antihypertensive agents with MRI white matter findings and with Modified Mini-Mental State Examination in older adults. AB - OBJECTIVES: To examine the association of antihypertensive regimen with magnetic resonance imaging (MRI) white matter hyperintensity and with cognitive impairment in older adults. DESIGN: Cross-sectional study. SETTING: The Cardiovascular Health study, an observational prospective cohort study of risk factors for coronary heart disease and stroke in men and women 65 years of age and older. PARTICIPANTS: 1268 men and women with pharmacologically treated hypertension. MEASUREMENTS: Information on medication use, medical history, and health habits was collected at clinic examinations. Participants completed the Modified Mini Mental State Examination (3MS) and underwent MRI examination. Without clinical information, study neuroradiologists assigned an overall grade of white matter signal intensity on MRI on a scale from 0 (no findings) to 9 (extensive findings). RESULTS: Adjusted mean white matter grade was higher for users of calcium channel blockers (2.59, P = .007) and users of loop diuretics (2.60, P = .015) than for users of beta blockers (2.12). The association was present for both dihydropyridine and non-dihydropyridine calcium channel blockers. Adjusted mean 3MS scores were lower for users of calcium channel blockers (89.6, P < .002), especially dihydropyridines, and users of loop diuretics (89.7, P < .006) than for users of beta blockers (92.3). No statistically significant association could be shown for users of other drug regimens, including thiazides and ACE inhibitors. CONCLUSION: In this study, users of antihypertensive regimens which included calcium channel blockers or loop diuretics had more severe white matter hyperintensity on MRI and worse performance on 3MS than users of beta blockers. PMID- 9400551 TI - Cerebral white matter changes (leukoaraiosis), stroke, and gait disturbance. AB - OBJECTIVES: Leukoaraiosis, a radiological change of cerebral white matter thought to be caused by ischemia, is associated with gait disturbance. However, because of concomitant stroke and cerebral atrophy, the clinical relevance of leukoaraiosis is uncertain. We, therefore, sought to determine if leukoaraiosis is a predictor of gait disturbance after accounting for cerebral atrophy and stroke in patients with a high prevalence of cerebrovascular disease. DESIGN: Cross-sectional observational study. SETTING: Neurology service (inpatient and outpatient) of a Department of Veterans Affairs Hospital. PARTICIPANTS: Consecutive sample of 130 patients, 127 men and three women. MEASUREMENTS: The findings of a gait scale were correlated to vascular risk factors, neurological examination as quantified by the NIH stroke scale and supplemental motor scale, and to brain CT findings. Brain CT scans were rated for leukoaraiosis, cerebral infarction, and cerebral atrophy. RESULTS: Gait disturbance was more frequent and more severe in subjects with leukoaraiosis, of whom 31% had mild and 49% moderate/severe gait disturbance compared with 27% with mild and 12% with moderate/severe gait disturbance in subjects without leukoaraiosis (P < .001). Leukoaraiosis, cerebral atrophy, a history of multiple strokes, and weakness and ataxia of the legs were independent predictors of gait disturbance. The proportion and severity of leukoaraiosis increased with increasing gait disturbance in subgroups without leg deficit (P < .001) and without multiple strokes (P < .001), but no association with leukoaraiosis was shown in patients with leg deficit or a history of multiple strokes (P = .037 and P = .186, respectively). Gait disturbance was more severe when both leukoaraiosis and cerebral atrophy were present (P = .019). CONCLUSION: In our Veteran population, leukoaraiosis is an independent predictor of gait disturbance after accounting for stroke and cerebral atrophy. Although leukoaraiosis is a form of cerebrovascular disease, it appears to be most closely associated to gait disturbance in the absence of symptomatic stroke or leg deficit. PMID- 9400552 TI - Physical activity and the changes in maximal isometric strength in men and women from the age of 75 to 80 years. AB - OBJECTIVE: To research the natural changes in maximal isometric strength, over a period of 5 years, in men and women aged 75 at baseline, and to study the effect of everyday physical activity on strength alterations. DESIGN: A 5-year longitudinal study. SETTING: Exercise laboratory. PARTICIPANTS: The target group in 1989 was the total 75-year-old population of Jyvaskyla. One hundred one men (81%) and 186 women (75%) participated in baseline strength tests, and after 5 years, 55 men and 111 women (70% and 72% of the survivors) took part in the follow-up measurements. METHODS: Maximal isometric hand grip, arm flexion, knee extension, trunk flexion, and trunk extension forces were measured using dynamometers. Self-rated physical activity was recorded using a scale by Grimby (1986). Strength changes were compared between groups based on the amount of everyday physical activity: (1) remained active (AA, 24 men, 24 women); (2) remained sedentary (SS, 11 men, 43 women); (3) decreased activity (AS, 11 women); and (4) increased activity (SA, 32 women). AS and SA could be formed for women only because of the small number of men. All analyses were stratified by gender. MAIN RESULTS: The average percentage change in strength over 5 years among survivors varied from a 4% increase in knee extension strength observed in men and women to a 16% decrease in grip strength in women. The grip strength decrease was greater in women than men. The AA men maintained their trunk extension strength at a higher level than the SS men. Time by group interactions in men were not significant. In women, the rate of decline in AS was 32% in grip and 27% in elbow flexion strength, which was greater than in the other activity groups. The AA women retained their knee extension strength at a higher level than the other groups. Those who died before follow-up tests exhibited poorer strength test results at baseline. Physical activity decreased over follow-up. CONCLUSIONS: Strength alterations with age differed between muscle groups. Undertaking everyday physical activities such as household work, walking, and gardening, which are also the most common physically demanding activities of older people, may play an important role in maintaining strength at an adequate level for independent living. PMID- 9400553 TI - Effects of age, physical training, and physical fitness on coronary heart disease risk factors in older track athletes at twenty-year follow-up. AB - OBJECTIVE: To compare current coronary heart disease (CHD) risk factor values in older athletes with mid-life measures and to examine the associations between changes in CHD risk factors with aging, physical training, and physical fitness. DESIGN: Prospective study with three longitudinal evaluation points: initial (T1), 10-year (T2), and 20-year (T3). Subjects were selected because of their elite status in Masters track competition. SETTING: University and medical center laboratories. PARTICIPANTS: Participants were 60 to 92 years of age and included 21 of the initial 27 subjects. At T3, subjects were divided into three groups, based on physical activity levels: high intensity (H), remained elite in national and international competition (n = 9); moderate intensity (M) continued frequent rigorous endurance training but rarely competed (n = 10); and low intensity (L) greatly reduced their training volume and intensity (n = 2). MEASUREMENTS: Smoking history; family history of coronary or cerebrovascular disease; resting blood pressure; resting electrocardiogram (ECG); serum total cholesterol, plasma glucose; body weight, % body fat, body mass index, waist:hip ratio; training pace and mileage; maximal oxygen consumption VO2 max). MAIN RESULTS: Several risk factors (smoking, diabetes, obesity) were never present, and the prevalence of other risk factors (family history of cardiovascular disease, abnormal resting ECG) remained low through T3 (< or = 14% of subjects). Mean systolic and diastolic blood pressure remained low without medication, but diastolic blood pressure measurements had the greatest redistribution between evaluation periods of any risk factor (r = .16, P = .479, T1 to T2). Mean total cholesterol was lower at T2 (-13%, P = .005) and T3 (-14%, P = .019) compared with T1. Change in VO2 max was correlated with changes in body weight (r = -.44, P = .048) and % fat (r = -.52, P = .015) from T1 to T2, whereas age was correlated to changes in systolic blood pressure (r = -.61, P = .003) and total cholesterol (r = -.49, P = .023) from T2 to T3. CONCLUSIONS: The prevalence of CHD risk factors remained low, and mean risk factor values remained low and generally stable in older athletes who had maintained habitual exercise training. PMID- 9400554 TI - Hypokalemia and potassium excretion in stroke patients. AB - OBJECTIVES: To determine (1) the prevalence of hypokalemia (plasma potassium < or = 3.4 mmol/L) in a group of stroke patients in comparison with age- and sex matched groups of patients having sustained a myocardial infarction or having mild hypertension and (2) the association between plasma potassium concentration and stroke outcome. DESIGN: Observational study. PARTICIPANTS: A total of 421 consecutive stroke patients admitted to a teaching hospital, 150 consecutive patients 50 years or older with myocardial infarction admitted to the hospitals Coronary Care Unit, and 161 out-patients 60 years or older with borderline and established hypertension. MEASUREMENTS: All stroke and cardiac patients had plasma urea and electrolytes estimated within 2 hours of hospital admission; in the hypertensive group blood samples were taken in clinic. Stroke patients had blood pressure, stroke severity (Barthel score) and smoking status recorded. A sub-group of 61 stroke patients and all 79 hypertensive patients not taking antihypertensive medication had 24-hour urine electrolyte excretion measured. Outcome (independent, dependent, or dead) at 3 months post-stroke was established in 349 patients. RESULTS: Hypokalemia occurred more frequently in stroke patients than in patients with myocardial infarction (84 (20%) vs 15 (10%), P = .008) or patients with hypertension (84 (20%) vs 13 (8%), P < .001), even when patients taking diuretics were excluded from analysis (56 (19%) vs 12 (9%) of cardiac group, P = .014 and 56 (19%) vs 4 (5%) of hypertensive group, P = .005, respectively). 24-hour urine excretion of potassium and the potassium:creatinine ratio was lower in stroke patients than in hypertensive patients (41 +/- 21 vs 62 +/- 25 mmol/24 hour, P = .001, 5.5 +/- 2.2 vs 7.4 +/- 2.6 mmol/24 hour, P = .001, respectively). On survival analysis, a lower plasma potassium on admission to hospital was associated with an increased chance of death, independent of age, stroke severity, history of hypertension, blood pressure level, or smoking history (hazard ratio 1.73 (95% CI: 1.03-2.9) for a 1 mmol/L lower plasma potassium concentration). CONCLUSIONS: Hypokalemia post stroke is common and may be associated with a poor outcome. PMID- 9400555 TI - The effect of specific medical conditions on functional decline. AB - OBJECTIVES: To examine how functional status among older community-dwelling residents differs over time between those with and those without specific medical conditions. DESIGN: Prospective cohort study. PARTICIPANTS: A total of 1060 community-dwelling Massachusetts residents aged 65 or older who were not totally functionally dependent at baseline assessment. MEASUREMENTS: Functional status, five medical conditions (heart problem, arthritis, diabetes, cancer, and stroke), and the total number of these five medical conditions. Assessments were done at baseline and at two annual follow-ups. RESULTS: Adjusted repeated measures analysis of covariance revealed a time difference (P < .001) for all five medical conditions and group differences for diabetes (P = .006) and stroke (P < .001). Functional abilities declined over time and those with specific medical conditions were more impaired initially, but the rate of decline did not significantly differ from those free of the condition. The presence of each additional medical condition resulted in additional impairment (P < .001), but the rate of decline over time did not differ by number of medical conditions. CONCLUSIONS: Efforts to reduce or prevent the development of specific medical conditions are essential to maintaining functional independence of older people as well as to reducing use of supportive services and admission rates to nursing homes. Particular attention should be directed toward preventing stroke since its consequences are the most functionally disabling. PMID- 9400556 TI - The synergy of low lung function and low body mass index predicting all-cause mortality among older Japanese-American men. AB - OBJECTIVE: To assess the joint characteristics of low standardized weight and compromised pulmonary function in predicting all-cause mortality. DESIGN: A population-based, prospective cohort study. SETTING: Oahu Island, Hawaii. PARTICIPANTS: Surviving Japanese-American men of the Honolulu Heart Program cohort, 71 to 93 years of age (N = 3059). MEASUREMENTS: Body mass index (BMI weight in kilograms/square of height in meters) and 1-second forced expiratory volume (FEV1) as a percentage of age- and height-predicted FEV1 from the 1991 to 1993 examination of the cohort. Mortality data derived from the ongoing tracking of deaths of the cohort. Relations of selected risk factors among joint levels of BMI (< or = 21, > 21 to < 25, > or = 25 kg/m2) and percent predicted FEV1 (< or = 70%, > 70%) were determined. The impact of these covariates on relations between joint BMI/percent predicted FEV1 levels and subsequent all-cause mortality was assessed. RESULTS: The highest age-adjusted mortality rate (91.9 deaths per 1000 person-years) was noted among men characterized by the joint conditions of percent predicted FEV1 < or = 70% and BMI < or = 21 kg/m2. This rate was 4.0 times the mortality rate of a "healthy" reference group characterized by percent predicted FEV1 > 70% and 21 < BMI < 25 kg/m2. This rate ratio is attenuated to 3.2 upon statistical control for measures of current and past smoking behavior. Among the three strata of BMI, statistical interaction is reflected in a heterogeneity of mortality rate differences (49.7, 21.8, -9.6 deaths/person-year, respectively) and rate ratios (2.18, 1.98, .66, respectively) comparing men with percent predicted FEV1 < or = 70% to > 70%. CONCLUSION: Joint loss of pulmonary function and relative weight is predictive of subsequent all-cause mortality in excess of additive or multiplicative effects of each condition separately. Smoking behavior may contribute to this observation. PMID- 9400557 TI - Mortality over four years in SHEP participants with a low ankle-arm index. AB - OBJECTIVES: To assess the risk of total and cardiovascular mortality in older adults with systolic hypertension and with a low ankle-arm index (AAI) as a marker of subclinical peripheral arterial disease (PAD). DESIGN: Prospective observational study PARTICIPANTS: A subgroup of 1537 participants in the Systolic Hypertension in the Elderly Program (SHEP) were screened for lower extremity arterial disease using the AAI. Participants were evaluated at 4 years to determine vital status and cause of death. Total and cardiovascular disease (CVD) mortality rates were assessed in relationship to clinical CVD at baseline, cardiovascular risk factors and the presence of a low AAI (subclinical PAD). RESULTS: Total mortality rates increased as the AAI decreased in those with and without clinical CVD at baseline. In those without clinical CVD at baseline, the presence of an AAI < or = .9 was associated with an age-adjusted relative risk (RR) of 3.00 for total mortality in men and 2.67 in women. Results were similar for CVD mortality and persisted after adjustment for cardiovascular risk factors including the presence of an abnormal electrocardiogram. CONCLUSIONS: A low ankle arm-index predicted a two to three-fold increase in total and cardiovascular mortality in older adults with systolic hypertension of risk for incident cardiovascular disease. In this study of older adults with systolic hypertension, 19.7% of the participants had subclinical PAD. Risk factor modification could be targeted to older adults based on markers of asymptomatic atherosclerosis. PMID- 9400558 TI - The impact of kyphosis on daily functioning. AB - OBJECTIVES: To determine whether moderate or severe kyphosis is associated with decrements in physical function, especially mobility. DESIGN: Cross-sectional analysis of a cohort study. SETTING: The Johns Hopkins Functional Status Laboratory, a multidisciplinary, standardized, quantitative assessment center. PARTICIPANTS: A total of 231 community-dwelling volunteers aged 59 and older who participated in a 1-day evaluation. MEASUREMENTS: Age, gender, self report of physical function, standardized measurement of: kyphosis (both qualitatively clinical criteria and quantitative assessment), time to walk 5 meters (0.1 seconds), and time to climb a flight of stairs (0.1 seconds) at usual pace. RESULTS: Using multivariate step-wise regression analysis, the presence and severity of kyphosis, measured qualitatively, was independently associated with time to walk 5 meters and to climb a flight of stairs (P = .015, P < .001, respectively), adjusting for moderate-severe scoliosis, heart rate response to exercise, arthritis, vertigo, age, and gender. Similarly, quantitative kyphosis was associated independently with stair climb time (P = .005). Qualitative kyphosis was also associated with difficulty reaching (OR = 2.21 (95% CI: 1.14 to 4.29)) and difficulty performing heavy housework (OR = 1.64 (95% CI: 1.03 to 2.61)), adjusting for prior diagnosis of moderate-severe scoliosis, prior diagnosis of arthritis, age, and gender. CONCLUSION: Kyphosis, by both clinical and quantitative assessment, is associated with diminished function, especially performance of mobility tasks. This association should be verified prospectively. If predictive, the impact of kyphosis on physical function should be considered in osteoporosis prevention and treatment counseling. PMID- 9400559 TI - Differences in psychosocial and health correlates of major and minor depression in medically ill older adults. AB - OBJECTIVES: To compare the differences in correlates of different levels of depression in medically ill hospitalized older adults. DESIGN, SETTING, AND PARTICIPANTS: A consecutive series of 542 patients aged 60 or older admitted to the medical inpatient services of Duke Hospital underwent a structured psychiatric evaluation administered by a psychiatrist. MEASUREMENT: A wide range of demographic, social, psychiatric, and physical health data were collected, and associations with major and minor depression were assessed. RESULTS: Compared with patients without depression, those with major depression were more likely to have a history of prior episodes of depression, higher dysfunctional attitude scores, greater overall severity of medical illness, cognitive impairment, and symptoms of pain or other somatic complaints. Specific medical diagnosis was less important a predictor of major depression than overall severity of medical illness. Compared with patients without depression, those with minor depression were more likely to report non-health-related stressors during the year before hospital admission, have a diagnosis of immune system disorder, and have greater severity of medical illness. When major and minor depression were compared directly, on the other hand, no significant differences were observed except for history of depression, and that relationship was weak and present only when the etiologic approach to diagnosis was used. CONCLUSION: During hospital admission, certain psychosocial, psychiatric, and physical health characteristics of older medical patients place them at high risk for different levels of depression. Patients with major and minor depression resemble each other more than they do patients without depression. These findings may help clinicians better understand the causes of different types of depression in this setting and lead to improved diagnosis and treatment. PMID- 9400560 TI - Older women and hormone replacement therapy: factors influencing late life initiation. AB - OBJECTIVE: To describe factors associated with initiation of hormone replacement therapy (HRT) by older women. DESIGN: A cross-sectional study of 671 randomly selected women aged 65 to 80 who participated in a larger telephone survey on preventive health behaviors. SETTING: A large health maintenance organization (HMO) in Seattle, Washington. PARTICIPANTS: Of the 521 women who responded (78%), 51 had begun taking HRT at age 60 or older and were identified as initiators. Women who had never used HRT or past users who had begun HRT before age 60 were classified as noninitiators (n = 362). Current users who started HRT before age 60 (n = 108) were excluded. MEASUREMENTS: Sources included the telephone survey, automated HMO pharmacy data, and HMO utilization and provider databases. RESULTS: Initiators were similar to noninitiators with respect to age, marital status, education, and health status. Initiators were more likely to have had a hysterectomy at age 60 or later than noninitiators. Sixty-two percent of the non initiators said they had received no information about the benefits of HRT from their providers compared with 18% of initiators. HRT initiation was associated with belief in prevention benefits of HRT for fractures and cardiovascular disease and with reported encouragement from the physician to use HRT. CONCLUSIONS: Other than hysterectomy status, there were few sociodemographic or health characteristics that markedly distinguished older initiators from noninitiators. Our findings show the importance of physician counseling in an older woman's decision to initiate HRT. PMID- 9400561 TI - Antidepressant use over time in a rural older adult population: the MoVIES Project. AB - OBJECTIVE: To examine the use of antidepressant drugs over time among community based older persons. DESIGN: A longitudinal community study with four approximately biennial data collection waves (1987-1996). SETTING: A low socioeconomic status rural older community-based population in Southwestern Pennsylvania. PARTICIPANTS: A total of 1681 individuals with a mean age of 72.9 years at study entry, MEASUREMENTS: Antidepressant drug use, demographics, and health services utilization by self-report. RESULTS: Antidepressant use was reported by less than 5% of the population during all four waves. It was associated with female gender, use of mental health services, presence of five or more depressive symptoms, and use of five or more prescription drugs, but not with age. During the four waves, tricyclics accounted for 84.6%, 85.3%, 78.4%, and 45.5% of total antidepressants used, whereas selective serotonin reuptake inhibitors (SSRIs) accounted for 2.6%, 11.8%, 8.1%, and 36.4%. CONCLUSIONS: Overall, our data on antidepressant use in this rural older population mirror national trends away from tricyclics and towards SSRIs. Our findings also suggest underutilization of mental health services and antidepressant drugs in this population. PMID- 9400562 TI - Preserved antilipolytic insulin action is associated with a less atherogenic plasma lipid profile in healthy centenarians. AB - OBJECTIVE: Recent studies have demonstrated that centenarians have a preserved glucose tolerance and insulin action and a more favorable body composition and fat distribution than aged subjects. The strong relationship among glucose tolerance, insulin action, plasma lipid concentration, and lipoprotein metabolism would lead to the hypothesis that healthy centenarians may also have a less atherogenic profile than aged subjects less than 100 years old. DESIGN: Investigation of the relationship between insulin action and lipid metabolism in healthy centenarians. PARTICIPANTS: Fifty-six subjects were categorized into three groups: Adults (< or = 50 years old; n = 20); Aged (> or = 75 years old; n = 22); Centenarians (> or = 100 years old; n = 14). The latter represented a select group of individuals free of major age-related diseases. MEASUREMENTS: Anthropometric measurements were made in all subjects, fasting blood samples were drawn for metabolite determinations, and an euglycemic glucose clamp was performed. RESULTS: Compared with aged subjects, healthy centenarians appeared to have a less atherogenic plasma lipid profile. Fasting plasma LDL cholesterol (2.4 +/- 0.6 vs 3.7 +/- .6 mmol/L P < .010) was significantly higher in aged subjects than in centenarians, whereas fasting plasma HDL cholesterol (1.0 +/- 0.4 vs 1.7 +/- .4 mmol/L P < .005) had an opposite trend. In centenarians, insulin-mediated glucose uptake was greater (34.6 +/- 0.5 vs 23.3 +/- .05 mumol/Kg FFM x min P < .010) than in aged subjects and correlated with fasting plasma triglycerides, FFA, LDL, and HDL cholesterol, Apo B, and Apo A1 concentrations. Finally, insulin infusion suppressed plasma FFA concentration in similar ways in adults and centenarians. CONCLUSION: Our study demonstrates that centenarians have a less atherogenic plasma lipid and lipoprotein profile than aged subjects. PMID- 9400563 TI - Development and validation of a clinical prediction rule for prolonged nursing home residence after hip fracture. AB - OBJECTIVES: To develop and validate a clinical prediction rule for nursing home residence 6 months after a hip fracture. DESIGN: Two prospective cohort studies, a development study (DS) and a validation study (VS). SETTING: The DS included hip fracture patients admitted to 92 rehabilitation units or skilled nursing facilities; the VS included hip fracture patients from 11 integrated healthcare systems. PARTICIPANTS: A total of 344 community-dwelling hip fracture patients aged 65 and older participated in the DS; 239 similar patients were enrolled in the VS. INTERVENTION: None. MEASUREMENTS: The acute hospital record, nursing evaluations, and patient questionnaires provided information about demographics, physical and neuropsychological function, and comorbidity. Residence 6 months after fracture was determined by phone interview. Multivariate analysis identified predictors for a risk score to assess the likelihood of nursing home residence. RESULTS: 18.7% of patients in the DS resided in nursing homes 6 months after hip fracture. The four independent risk factors for institutionalization were (1) being unmarried (OR = 6.7 [95% CI 2.4 to 19]), (2) incontinence (OR = 2.3 [CI 1.2 to 4.7]), (3) dependence in ambulation (OR = 5.0 [CI 2.1 to 12.3]), and (4) cognitive impairment (OR = 6.6 [CI 3.3 to 13.2]). Of patients with all four risk factors, 73.2% were institutionalized at 6 months, compared with 0% of patients with no risk factors. In the VS, 6.1% of patients resided in nursing homes after 6 months, with a range from 50.0% of patients with four risk factors to 0% of those with no risk factors. Areas under receiver-operating characteristic curves for the prediction rule were 0.84 +/- .03 in the DS, and 0.81 +/- .06 in the VS. CONCLUSION: A clinical prediction rule using four easily measurable characteristics can identify individuals at high or low risk of nursing home residence 6 months after hip fracture. PMID- 9400564 TI - Geropsychiatric restraint use. AB - OBJECTIVES: To investigate predictors and reasons for restraint use with geropsychiatric patients. DESIGN: A prospective, correlational study. SETTING: The geriatric unit of an acute-care psychiatric hospital. PARTICIPANTS: Twenty one staff nurses and 131 patients admitted consecutively over a period of 6 months. MEASUREMENTS: Disruptive behaviors were measured with the Nursing Home Behavior Problem Scale (NHBPS), cognitive function was measured with the Mini Mental State Examination (MMSE), mobility was measured with a Functional Mobility Screen (FMS), and reasons for restraint use were obtained with a questionnaire completed by nurses. RESULTS: Patients with a diagnosis of dementia, impaired mobility, or behavioral problems were more likely to be restrained. The most frequent reasons given by staff for restraint use were an unsteady gait and a risk of falling. The incidence of restraint use was 27.1%. CONCLUSIONS: The use of restraint with geropsychiatric patients may be more common than previously reported and requires further investigation. PMID- 9400565 TI - Health care for older persons, a country profile: Italy. PMID- 9400566 TI - A model for nurse case-managed home care using televideo. PMID- 9400567 TI - Antihypertensive drugs, brain structure, and cognitive function: more research is necessary. PMID- 9400568 TI - High tech comes to high touch. PMID- 9400569 TI - Do older individuals need more than usual physical activities to maintain muscle strength and function? PMID- 9400570 TI - Apolipoprotein E level in cerebrospinal fluid increases with aging. PMID- 9400571 TI - Increased oxidative damage in lymphocytes of Alzheimer's disease patients. PMID- 9400572 TI - Renal failure in older people: a preventable hospital admission and ageism in decision making. PMID- 9400574 TI - Size and structure of the thyroid in old age. PMID- 9400573 TI - Nutrition and immunity in older people. PMID- 9400575 TI - Squalor syndrome. PMID- 9400576 TI - Interviewing adolescents. AB - Incorporating a functional approach within the traditional structure of the medical interview allows for improved communication with adolescent patients. Using these techniques results in improved patient satisfaction, more accurate psychosocial diagnosis, and better adherence to treatment recommendations. Many physicians think that using patient-centered interviewing techniques is time consuming and inefficient, but, in fact, a higher quality and quantity of information usually is obtained per unit time using these techniques. An additional benefit is that this approach is more satisfying for the physician; the enhancement of the therapeutic relationship works both ways. Adolescents become more interesting as people; communication barriers become challenges to be creatively solved rather than annoyances to complain about. Adolescents need and deserve high-quality health care to grow into physically and mentally healthy adults, and effective physician-patient communication skills are the key to delivering that care. PMID- 9400577 TI - Integrating comprehensive adolescent preventive services into routine medicine care. Rationale and approaches. AB - Relying on therapeutic interventions to address health problems after they occur has proven costly and does not address the need to reduce the number of youth who develop these health problems. Primary care physicians have an important role to play in promoting adolescent health through a strategy of providing health guidance to adolescents and parents, screening, and promoting immunizations. Reducing the health risk behaviors of adolescents is a challenge that is best accomplished with the support of other preventive initiatives. Clinical preventive services should complement and reinforce preventive efforts in schools (i.e., comprehensive school health programs) and communities (i.e., mass media campaigns and health regulations). GAPS recommendations developed by the AMA and recommendations from other groups provide a model for organizing the content and delivery of comprehensive preventive services for adolescents. Physicians and other primary care health providers may use these recommendations to expand the quantity and quality of preventive services they offer to adolescents. Additional information about preventive services and GAPS, including a complete list of the recommendations, dates for future GAPS training, a description of the materials developed to help implement preventive services, other national efforts in adolescent preventive services, and current articles in scientific literature, can be reviewed on the AMA web site (http:/(/)www.AMA-Assn.Org/Adolhlth/Adolhlth+ ++.htm). PMID- 9400578 TI - The pediatrician and the sexually active adolescent. Sexual activity and contraception. AB - Sexuality and its resultant consequences continue to be major issues for adolescents and for those who provide their health care. This article discusses current sexual behavior in adolescents and describes the various forms of hormonal contraception that sexually active adolescents should use. PMID- 9400579 TI - The pediatrician and the sexually active adolescent. Treatment of common menstrual disorders. AB - Menstrual disorders in adolescents are a common medical problem. For young adolescents, the onset of menses is a time of dramatic physical and psychological change. Providing education to young women regarding the normal process of puberty helps ease some of the anxiety regarding this change. Health professionals can participate in the education of young women by defining the normal process of puberty and menstruation. Menstrual disorders, such as dysmenorrhea, irregularities in menstrual flow, and premenstrual symptoms, can be effectively diagnosed and treated in the adolescent population. PMID- 9400581 TI - Office approach to drug abuse prevention. AB - The onset of tobacco, alcohol, and other drug use generally occurs during adolescence. While many teens experiment with these substances, a significant number use them to the point that their behavior interferes with school, family, social relationships, and general productivity. It is the pediatrician's responsibility to identify young people who are at risk for the subsequent use of these substances and initiate a carefully designed prevention program in the office setting. PMID- 9400580 TI - The pediatrician and the sexually active adolescent. A primer for sexually transmitted diseases. AB - Sexual activity is a common practice among young adolescents, placing them at high risk for STDs, many of which have long-term consequences. Early diagnosis and treatment are essential to limit both the consequences and the spread of these infections. The clinician has a responsibility to the adolescent patient to recognize and treat these diseases. Using history and physical examination, the clinician should be able to determine an adolescent's risk for an STD, and, based on this risk, undertake the appropriate evaluations. Patient treatment, follow up, and management of sex partners are then guided by the results of either presumptive or definitive diagnostic tests. PMID- 9400582 TI - The rebellious adolescent. Evaluation and management of oppositional and conduct disorders. AB - A wide variety of management options are available to the primary care physician who is presented with a rebellious adolescent. After a careful assessment, the clinician and other health care professionals can choose a diverse combination of interventions: individual therapy, family therapy, youth-centered programs, community-centered programs, psychopharmacology, and others. Rebellious adolescents need access to comprehensive medical and mental health care, academic education (including sexuality education), and full employment opportunities. Primary care physicians can play a vital and sometimes pivotal role coordinating services and helping parents, school personnel, and therapists work with these youth. Even when dealing with the very difficult and resistant group of youth with CD and ODD, optimism for improvement should always be maintained by the clinician. PMID- 9400583 TI - Juvenile mood disorders and office psychopharmacology. AB - Mood disorders afflict pediatric patients, cause significant impairment, and interfere with normal development. Increasingly, pediatricians are called on to assess and collaborate with mental health practitioners in medicating children and adolescents with mood disorders. Approaching the juvenile with a primary emphasis on clarifying the diagnoses, determining environmental antecedents and sequelae, and investigating suicide risk enables the pediatrician to institute appropriate treatment. Despite limited data from controlled studies, psychotherapy often is used for mild to moderate depression. Pharmacotherapy is indicated in cases unresponsive to psychotherapy and in severe or suicidal cases. First-line pharmacotherapy for depressed adolescents is usually an SRI followed by the atypical or TCA antidepressants. Bipolar disorder typically requires an aggressive medication regimen, including anticonvulsants, lithium, or a combination, as well as environmental modifications. With severe, difficult, or refractory cases, mental health consultation is recommended to clarify diagnoses and to provide psychotherapy and medication input. PMID- 9400584 TI - Acne. What every pediatrician should know about treatment. AB - Acne is one of the most common and easily treated diseases of adolescents. Scarring can be prevented in most cases with early and vigorous treatment. But such treatment requires patience, skill, and a commitment to good counseling. PMID- 9400585 TI - The preparticipation sports examination. AB - Although not a comprehensive health evaluation, the PSE is viewed as such by many parents and adolescents. In this regard, these athletes are getting a lower standard of care than recommended; however, the mechanism wherein the PSE can become a cost-effective comprehensive examination has not been established. The PSE is one method of injury prevention in that it is designed to identify medical conditions that would be worsened by participation in exercise and sports. Pediatricians should use this examination as a quality control point during the year to assess how medical conditions and musculoskeletal injuries have been diagnosed and managed in the context of sports. PMID- 9400586 TI - What every pediatrician should know about infectious mononucleosis in adolescents. AB - Infectious mononucleosis (IM) is one of the most common diseases occurring during adolescence. Appreciation of IM's varied clinical presentations, its differential diagnosis, and the difficulties involved in making the laboratory diagnosis will enable clinicians to treat teenagers more effectively in their office practices. PMID- 9400587 TI - Psychosomatic problems and stress in adolescence. AB - Psychosomatic problems are common in adolescents, and stress frequently plays a role in their development and maintenance. Armed with an understanding of the stressors experienced by adolescents, the individual's vulnerabilities and competencies and their level of social support, the physician can systematically assess each of these factors. Once the assessment is complete, a management plan can be formulated to address the particular psychosomatic problem. Symptom relief, stress reduction, and promotion of competence are important interventions that can be initiated by the primary care physician. When referrals are made for counseling and other stress management techniques, the primary care physician should maintain contact with the patient and family and remain an integral part of the management team. Incorporating brief discussions about the potential role of stress in health and illness into anticipatory guidance sessions may also help prevent the development of psychosomatic problems in adolescents. PMID- 9400588 TI - Evaluation of hematuria, proteinuria, and hypertension in adolescents. AB - Signs or symptoms of renal disease in adolescents deserve prompt attention and appropriate evaluation. Adolescents are susceptible to a variety of urinary tract disorders. The key issue in the evaluation of hematuria or proteinuria in adolescents is the existence of concomitant signs of renal disease. For isolated hematuria or proteinuria, demonstration of persistence and a reasoned evaluation are in order. Hypertension in adolescents must be carefully documented and, when present, considered seriously. The fact that most teens with persistent elevated blood pressures have essential hypertension is still a great concern because for most of these adolescents the hypertension will be lifelong and, if left untreated, can be associated with significant morbidity and mortality in the adult years. PMID- 9400589 TI - Common cardiac diseases in adolescents. AB - Adolescent patients frequently present with symptoms potentially referable to the cardiovascular system; however, patients rarely have true cardiovascular disease. In many patients, the history is the key to determining whether additional testing or referral is necessary. Even though the history, physical examination, and preliminary laboratory data may indicate a benign cause, occasionally a referral to a cardiologist is necessary because of patient or parental anxiety. Patients with true cardiac pathology should be followed by a cardiologist in conjunction with the primary care physician. PMID- 9400590 TI - Immunity to rotavirus in T cell deficient mice. AB - Rotavirus infection was studied in adult nude mice (BALB/c background), alpha beta or gamma delta and alpha beta/gamma delta T cell receptor (TCR) knockout (-/ ) mice (C57BL/6 and C57BL/6 x 129 backgrounds), and SCID mice (C57BL/6 background). The gamma delta TCR -/- mice cleared infection just like control mice. All of the nude mice, alpha beta, and alpha beta/gamma delta TCR -/- mice cleared primary rotavirus infection, with a short delay, compared to immunocompetent control mice and developed a rotavirus-specific intestinal IgA measured by ELISA. Elispot analysis with spleen and lamina propia cells showed that the virus-specific intestinal IgA response in immunocompetent C57BL/6 mice was similar to the gamma delta TCR -/- mice and 7- to 60-fold higher than in the alpha beta TCR -/- and alpha beta/gamma delta TCR -/- mice. Likewise, the response of nude +/- mice was 20 times greater than that of nude -/- littermates. While the intestinal IgA antibodies of C57BL/6 mice, gamma delta TCR -/- mice, and nude +/- mice recognized insect cells infected with recombinant baculovirus expressing rotavirus VP6 and VP4 proteins, those of the alpha beta TCR -/-, alpha beta/gamma delta TCR -/-, and nude -/- mice recognized only VP6. Immunocompetent C57BL/6 mice depleted of CD4+ T cell developed similar levels of rotavirus specific intestinal IgA as the alpha beta TCR -/- mice, suggesting that this T cell-independent IgA response is present in normal mice. In contrast to previously published results with BALB/c SCID and RAG 2 -/- (C57BL/6 x 129 background) mice, all of which become chronically infected with murine rotavirus, 40% of the C57BL/6 SCID mice cleared primary rotavirus infection. These results suggest that both a T cell-independent antibody response and innate mechanisms can contribute to immunity to murine rotavirus and show that gamma delta T cells are not necessary for efficient clearance of primary rotavirus infection in mice. PMID- 9400591 TI - Apoptosis of CD4+ and CD19+ cells during human immunodeficiency virus type 1 infection--correlation with clinical progression, viral load, and loss of humoral immunity. AB - Enhanced rates of programmed cell death (apoptosis) have been detected in T cells and B cells from human immunodeficiency virus type 1 (HIV-1)-infected individuals. To evaluate the possible relevance of this event to HIV pathogenesis and disease progression, apoptosis in CD4+ T lymphocytes and CD19+ B lymphocytes, viral load, and neutralizing antibody titers were assayed in HIV-1-infected slow progressors and progressors. A correlation was found between progressive disease and apoptosis of CD4+ T cells. The extent of apoptosis in CD4+ cells was similar in slow progressors and seronegative control subjects. By contrast, we found elevated levels of B-cell apoptosis in all HIV-1-infected individuals compared with seronegative control subjects, with a tendency toward increased levels of apoptosis with progressive disease. Apoptosis in CD4+ T cells and CD19+ B cells correlated with viral RNA levels in plasma. Furthermore, higher rates of B-cell apoptosis were observed in individuals with poor neutralizing activity against a panel of six clinical HIV-1 isolates. From these results we conclude that the extent of apoptosis in cultured CD4+ cells and CD19+ cells appears to parallel the decline in CD4 cell counts in infected individuals. The finding of a relation between apoptosis in B cells and poor neutralizing capacity suggests that apoptosis may be related to loss of immune function. A role for apoptosis in the pathogenesis of AIDS is also supported by the strong correlation between viral load and rates of apoptosis in CD4+ T cells. PMID- 9400592 TI - Interferon-gamma protects against herpes simplex virus type 1-mediated neuronal death. AB - Host inflammatory mediators, such as interferons, play a protective role in infection, but the mechanism is undefined and may differ between tissue compartments. To determine whether interferon-gamma (IFN-gamma) elicitation prevents destructive encephalitis in herpes simplex virus type 1 (HSV-1) infection of the central nervous system, IFN-gamma-knockout (GKO) mice were challenged intravitreally with HSV-1 strain F, inciting infection of the eyes and the brain. Indeed, the GKO mice showed encephalitis with ataxia, whereas nontransgenic controls remained asymptomatic. Morphology and digoxigenin labeling of DNA fragments revealed increased apoptosis in the brains of GKO mice compared with controls, although viral replication was not influenced at early stages of infection. Greater numbers of apoptotic cells in the brains of GKO mice correlated with neurological symptoms, as well as lower expression of the protective protooncogene bcl-2. Thus, IFN-gamma inhibits apoptosis, affording neuronal protection from destructive encephalitis during viral infection of the central nervous system. PMID- 9400593 TI - Host range and cytopathogenicity of the highly attenuated MVA strain of vaccinia virus: propagation and generation of recombinant viruses in a nonhuman mammalian cell line. AB - Modified vaccinia virus Ankara (MVA), attenuated by over 500 passages in primary chick embryo fibroblasts (CEF), is presently being used as a safe expression vector. We compared the host ranges of MVA and the parental Ankara strain in CEF and 15 permanent cell lines. The cells could be grouped into three categories: permissive, semipermissive, and nonpermissive. For MVA, the permissive category consisted of primary CEF, a quail cell line derived from QT6, and the Syrian hamster cell line BHK-21. Only in BHK-21 cells did the virus yield approach that occurring in primary CEF. The semipermissive category included two African green monkey cell lines: BS-C-1 and CV-1. The nonpermissive category for MVA consisted of three human cell lines HeLa, 293, and SW 839; one rhesus monkey cell line FRhK 4; two Chinese hamster cell lines CHO and CHL; one pig cell line PK(15); and three rabbit cell lines RK13, RAB-9, and SIRC. The grouping for MVA with a restored K1L host range gene was similar except for the inclusion of RK13 cells among permissive lines. The grouping for the Ankara strain, however, was quite different with more permissive and semipermissive cell lines. Nevertheless, in cells that were permissive for MVA, the virus replicated to higher levels than Ankara, consistent with both positive and negative growth elements associated with the adaptation of MVA. The cell lines were also characterized according to their susceptibility to MVA-induced cytopathic effects, expression of a late promoter regulated reporter gene by an MVA recombinant, and stage at which virion morphogenesis was blocked. Finally, the permissive BHK-21 cell line was shown to be competent for constructing and propagating recombinant MVA, providing an alternative to primary CEF. PMID- 9400594 TI - Phylogenetic analysis of simian T-lymphotropic virus Type I (STLV-I) in common chimpanzees (Pan troglodytes): evidence for interspecies transmission of the virus between chimpanzees and humans in Central Africa. AB - Serum and peripheral blood leukocytes from the chimpanzees (Pan troglodytes) of the colony of the Laboratory of Central Nervous System Studies, National Institute of Neurological Disorders and Stroke, NIH, were tested for the presence of STLV-I-specific antibodies and proviral DNA. Antibodies were determined by gelatin particle agglutination and Western blot (WB) assays utilizing HTLV-I antigens. Proviral DNA was detected by four PCR assays targeting three different regions of STLV-I genome: the fragments of the env and pol genes and LTR. Twenty of twenty-two DNA samples from WB-positive animals were PCR positive. None of the DNA samples from WB-negative (n = 5) and WB-indeterminate (n = 4) animals was PCR positive. The results of the nested and double nested env PCR tests were fully concordant; the seminested LTR PCR test was much less sensitive. The DNA sequences from the env (483 bp) and the pol (200 bp) genes and LTR (705 bp) were determined for six, two, and two chimpanzee STLV-I isolates, respectively. Phylogenetic analysis revealed that chimpanzee STLV-I isolates can be attributed to three clades. The first of these clades (SS-PTR1/CSA) included STLV-I isolates from the chimpanzees and West African subspecies of African green monkeys (Cercopithecus a. sabaeus). The other clades (S-PTR2 and S-PTR3) included STLV-I isolates only from chimpanzees. However, both S-PTR2 and S-PTR3 clustered together with Central African HTLV-I comprising the human/simian clade (HS HSA/PTR). This pattern of phylogenetic clustering suggests that interspecies transmission of STLV-I occurred between chimpanzees and African green monkey subspecies as well between chimpanzees and human populations in Central Africa. PMID- 9400595 TI - Interactions between Tat of HIV-2 and transcription factor Sp1. AB - Tat of HIV-2 (Tat-2) requires host cellular factors for optimal function. We show that transactivation by Tat-2 of the HIV promoter requires cis-acting binding sites for Sp1 or Sp1 brought to the promoter via a heterologous system. We demonstrate that an activation domain in Tat-2 consists of one of two potential alpha-helices in the amino-terminal region, the cysteine-rich region, and the core region and that this independent activation domain requires cis-acting Sp1 binding sites for function. Tat-2 interacts with Sp1 in in vitro binding assays, and these interactions require basic residues outside of the Tat-2 activation domain. The regions in Sp1 sufficient for functional synergy with Tat are the Sp1 activation domains, while the DNA-binding region is dispensable. Substitution mutations of a glutamine-rich region in one Sp1 activation domain, which eliminate interactions with a TBP-associated factor, also significantly decrease synergy with Tat. Thus, the functional synergy between Tat-2 and Sp1 localizes to domains in each activator that interact with components of the transcription complex. We suggest that these interactions, rather than direct Tat/Sp1 binding, result in highly processive RNA polymerase II complexes and full-length viral transcripts. PMID- 9400596 TI - Direct effect of type 1 human immunodeficiency virus (HIV-1) on intestinal epithelial cell differentiation: relationship to HIV-1 enteropathy. AB - Human immunodeficiency virus (HIV)-infected patients display severe impairments of gastrointestinal functions, including diarrhea and malabsorption, even in the absence of opportunistic infections. Since HIV-1 proteins and nucleic acids have been detected in several cell types of the intestinal mucosa, it has been postulated that HIV-1 itself could alter enterocytic functions. In the present study, we analyzed the effect of HIV-1 on the differentiation process of the epithelial intestinal cell clone HT-29-D4, which mimics the maturation of enterocytes along the crypt-villus axis of the small intestine. We found that HIV 1 infection impairs cellular differentiation (i) by affecting the barrier function of the epithelium, as evidenced by a decrease in the transepithelial electrical resistance, and (ii) by inhibiting the activity of one major glucose absorption function, i.e., sodium/glucose cotransport. At the morphological level, HIV-1 infection of HT-29-D4 cells was associated with the formation of lumina, which are representative of a defect in cellular organization. These morphofunctional perturbations induced by HIV-1 could be mimicked by nocodazole, a microtubule-disrupting agent. Correspondingly, HIV-1 exposure of HT-29-D4 cells evoked a massive disruption of microtubules, as revealed by alpha-tubulin indirect immunofluorescence staining. A similar effect was observed after incubation of the cells with either recombinant gp120 or a monoclonal antibody against galactosylceramide (GalCer), the intestinal receptor for HIV-1 gp120, suggesting that the effect of HIV-1 was mediated by the binding of gp120 to GalCer. Based on these data, we propose that HIV-1 may selectively alter enterocytic functions through a direct effect on the intracellular architecture of the cells. In contrast with previous theories for HIV-1 enteropathy, our data support the concept that HIV-1 may perturb intestinal functions without necessarily infecting intestinal epithelial cells. PMID- 9400597 TI - Liquefaction of Autographa californica nucleopolyhedrovirus-infected insects is dependent on the integrity of virus-encoded chitinase and cathepsin genes. AB - We examined the role of the Autographa californica nucleopolyhedrovirus (AcMNPV) encoded chitinase in virus pathogenesis in Trichoplusia ni larvae. In conjunction with the AcMNPV-encoded cathepsin, it promotes liquefaction of the host in the latter stages of infection. Insects infected with virus mutants lacking either the chitinase A gene (chiA) or cathepsin gene (cath) remained intact several days after death. However, if both viruses were used to infect insects, liquefaction of the host was restored. Chitinase was readily detected in AcMNPV-infected insects using a chitinase-specific antibody, but it was absent from insects infected with a chiA deletion mutant (AcchiA-). The chitinase was also detected in polyhedra purified from AcMNPV-infected insects but not in those from AcchiA-. However, polyhedra derived from a virus lacking an intact chiA were no less effective in initiating an infection in second instar T. ni larvae than those of the unmodified AcMNPV. It was also demonstrated that the virus chitinase retained high levels of activity between pH 3.0 and 10.0. In contrast, chitinases isolated from Serratia marcescens, although active under acidic conditions, rapidly lost activity above pH 7.0 illustrating that despite 57% sequence identity, the two proteins have distinct enzymic activities. PMID- 9400598 TI - Passage of HIV-1 molecular clones into different cell lines confers differential sensitivity to neutralization. AB - In this study, progeny viruses of four HIV-1 molecular clones were tested for sensitivity to neutralization following prolonged passage in peripheral blood mononuclear cells (PBMC) and MT-2, H9, and CEM T-lymphoid cell lines. Two of the viruses were able to establish persistent infection with no cytopathic effect in H9 and CEM cells. Such adaptation conferred increased sensitivity to neutralization by a panel of human sera obtained from HIV-1-infected asymptomatic individuals, by soluble CD4 and by monoclonal antibodies directed to a linear epitope in the V3 region (268-D) and a conformational epitope in the CD4 binding site of the envelope gp 120 (1.5e). Increased sensitivity to neutralization was paralleled by increased binding of these mAbs to native envelope glycoproteins and by increased binding capacity to CD4 expressed on the cell surface. Our results show that virus-host cell interactions are important in influencing sensitivity to neutralization of HIV-1. In primary PBMC or in cytopathic interactions in cell lines, like in MT-2 cells, envelope epitopes important for neutralization remain masked. In contrast, noncytopathic but productive virus host cell interactions may lead to an increased exposure of neutralizing epitopes and more efficient binding capacity to CD4 resulting in an increased sensitivity to neutralization. PMID- 9400599 TI - Mutations affecting the sensitivity of the influenza virus neuraminidase to 4 guanidino-2,4-dideoxy-2,3-dehydro-N-acetylneuraminic acid. AB - 4-Guanidino-2,4-dideoxy-2,3-dehydro-N-acetylneuraminic acid (4-guanidino Neu5Ac2en) specifically inhibits the influenza virus neuraminidase (NA) through interaction of the guanidino group with conserved Glu 119 and Glu 227 residues in the substrate binding pocket of the enzyme. To understand the mechanism by which influenza viruses become resistant to 4-guanidino-Neu5Ac2en, we investigated mutations at amino acid residues 119 and 227 in the influenza virus NA for their effects on this compound and on NA activity. The NA gene was cloned from the NWS G70c virus, and mutations were introduced at the codon for amino acid residue 119 or 227. All of the 13 mutants containing a change at residue 119 were transported to the cell surface, although their expression levels ranged from 68.2 to 91.3% of wild type. Mutant NAs that retained at least 20% of the wild-type enzymatic activity were tested for their sensitivity to 4-guanidino-Neu5Ac2en and found to be sevenfold less sensitive to this compound than was the wild-type NA. By contrast, only 6 of 13 mutants defined by modifications at residue 227 were transported to the cell surface, and those NAs lacked substantial enzymatic activity (9% of wild type, at most). These results suggest that only a limited number of resistant viruses arise through mutations at Glu 119 and Glu 227 under selective pressure from 4-guanidino-Neu5Ac2en and that the development of compounds which interact with 227 Glu more strongly than does 4-guanidino Neu5Ac2en may reduce the likelihood of drug-resistant viruses still further. PMID- 9400600 TI - Insights into the interaction between tRNA and primer binding site from characterization of a unique HIV-1 virus which stably maintains dual PBS complementary to tRNA(Gly) and tRNA(His). AB - Previously our laboratory constructed an HIV-1 which stably maintained a primer binding site (PBS) complementary to tRNA(His) by mutating the region of the provirus within U5 postulated to interact with the anticodon of tRNA(His) (J. Wakefield, S-M Kang, and C. D. Morrow, 1996, J. Virol, 70, 966-975). From the analysis of the virus obtained after long-term culture, we identified an unusual proviral DNA in which the U5-PBS region contained a dual PBS complementary to tRNA(Gly) and tRNA(His), respectively, separated by a 21-nucleotide intervening sequence. To determine if this U5-PBS region containing the dual PBS would give rise to an infectious virus, the mutant U5-PBS containing the dual PBS was subcloned into an infectious HIV-1 proviral clone, pHXB2; the resultant proviral DNA was designated as pHXB2(Gly-His). Transfection of pHXB2(Gly-His) into cells gave rise to infectious virus. Analysis of the U5-PBS region revealed that the virus stably maintained the dual PBS rather than revert back to the wild-type PBS. In addition to genomes with the PBS complementary to tRNA(Gly) and tRNA(His), proviral genomes were identified after extended in vitro culture which contained dual PBS complementary to tRNA(Gly) and tRNA(Phe). To determine which PBS could be used for reverse transcription, we utilized an endogenous reverse transcription/PCR method which could discriminate (based on molecular size of the products) between the minus strand DNA initiated from the two PBSs. The results of this assay demonstrated that either the PBS complementary to tRNA(Gly) or tRNA(His) could be used for the initiation of reverse transcription. The results of our study highlight the complex interrelationship between U5-PBS and primer tRNA required for positioning the tRNA at the PBS and provides new insights into how the tRNA primer used to initiate reverse transcription is selected. PMID- 9400601 TI - A glycine to alanine substitution in the paramyxovirus SV5 fusion peptide increases the initial rate of fusion. AB - Simian virus 5 fusion (F) protein mutant F-G3A, which contains a glycine-to alanine substitution at position 3 in the conserved hydrophobic fusion peptide at the N-terminus of the F1 subunit, has been shown previously to cause increased syncytium formation compared to wild-type (wt) F protein, when expressed using an SV40 recombinant virus vector system (C. M. Horvath and R. A. Lamb (1992) J. Virol. 66, 2443-2455). The wt F and the F-G3A proteins were expressed in eukaryotic cells using the vaccinia virus-bacteriophage T7 RNA polymerase (vac T7) expression system, and they showed similar cell surface expression levels as determined by flow cytometry. The final extent of fusion when the vac-T7 expression system was used was not found to be greatly different when examined with a reporter gene activation assay. However, the initial rate of fusion was found to be five- to sixfold higher for the F-G3A mutant protein than the wt F protein, when examined using a quantitative assay for lipid mixing based on relief of self-quenching of fluorescence of the lipid probe octadecyl rhodamine (R18). A microscopic fluorescent dye transfer assay also showed a much earlier spread of dye from R18-labeled red blood cells to the cells expressing the mutant F-G3A protein than the wt F protein. Thus, these data indicate that a single gly to-ala mutation in the fusion peptide domain, although not affecting the final extent of fusion, significantly increased the rate of fusion. Possible mechanisms for the increased rate of fusion are discussed. PMID- 9400602 TI - Analysis of a peptide inhibitor of paramyxovirus (NDV) fusion using biological assays, NMR, and molecular modeling. AB - To investigate the molecular mechanisms involved in paramyxovirus-induced cell fusion, the function and structure of a peptide with a 20-amino-acid sequence from the leucine zipper region (heptad repeat region 2) of the Newcastle disease virus fusion protein (F) were characterized. A peptide with the sequence ALDKLEESNSKLDKVNVKLT (amino acids 478-497 of the F protein) was found to inhibit syncytia formation after virus infection and after transfection of Cos cells with the HN (hemagglutinin-neuraminidase) and F protein cDNAs. Using an F protein gene that requires addition of exogenous trypsin for cleavage, it was shown that the peptide exerted its inhibitory effect prior to cleavage. The three-dimensional conformation of the peptide in aqueous solution was determined through the use of NMR and molecular modeling. Results showed that the peptide formed a helix with properties between an alpha-helix and a 3(10)-helix and that leucine residues aligned along one face of the helix. Side chain salt bridges and hydrogen bonds likely contributed to the stability of the peptide secondary structure. Analysis of the aqueous solution conformation of the peptide suggested mechanisms for specificity of interaction with the intact F protein. PMID- 9400603 TI - Interaction of endocytic signals from the HIV-1 envelope glycoprotein complex with members of the adaptor medium chain family. AB - The envelope glycoprotein (Env) complex of HIV-1 undergoes rapid internalization from the plasma membrane of human cells by virtue of a tyrosine-based endocytic signal (RQGYSPL, residues 704-710) in the cytosolic tail of the protein (J. F. Rowell et al., J. Immunol. 155, 473-488, 1995). Here we demonstrate that this tyrosine-based signal interacts with the mu 2 (medium) chain of the AP-2 clathrin associated adaptor, a protein complex involved in endocytosis of cell surface receptors. The same signal is also capable of interacting with two other members of the adaptor medium chain family, mu 1 and mu 3A, which are components of the AP-1 and AP-3 adaptor complexes, respectively. Interactions with mu 1 and mu 3A might be responsible for the targeting of the internalized envelope glycoprotein to lysosomes or to the basolateral plasma membrane of polarized epithelial cells. A second potential tyrosine-based signal (LFSYHRL, residues 760-766) also interacts with mu 1, mu 2, and mu 3A, although it is less important for internalization in vivo probably due to its position within the cytosolic tail. Overexpression of chimeric proteins having the HIV-1 Env cytosolic tail increases expression of the transferrin receptor on the cell surface, probably due to saturation of the cellular pool of mu 2 by the overexpressed proteins. These observations suggest that HIV-1 Env utilizes the protein sorting machinery of the host cells for internalization and sorting at various steps of the endocytic and biosynthetic pathways. PMID- 9400604 TI - Differential replication of ovine lentivirus in endothelial cells cultured from different tissues. AB - Blood-brain barrier dysfunction has been postulated to be important in the pathogenesis of HIV dementia. This study used an in vitro model of the blood brain barrier to determine the effects of ovine lentivirus (OvLV) infection on endothelial cells. The replication of two American OvLV isolates and two lcelandic OvLV isolates in pure cultures of endothelial cells isolated from brain was compared to replication in endothelial cells from adipose, lung, and aorta. Inoculation with the two American isolates resulted in 100 times greater reverse transcriptase (RT) activity in supernatant of the microvascular endothelial cells (brain, lung, and adipose) than in the macrovascular endothelial cells (aorta). Conversely, inoculation with the two lcelandic isolates resulted in 100 times higher RT activity in aortic, lung, and adipose endothelial cells than in the brain endothelial cells. Transmission electron microscopy of the brain capillary endothelial cells infected with the American isolates revealed polarized viral budding from the lateral cell membrane and a loss of tight junctions. Replication of OvLV in brain capillary endothelial cells could play a role in the pathogenesis of lentiviral encephalitis by altering blood-brain barrier integrity. PMID- 9400605 TI - Efficacy of replication-defective adenovirus-vectored vaccines: protection following intramuscular injection is linked to promoter efficiency in muscle representative cells. AB - To investigate the respective role of transduced cells in the induction of immune response following intramuscular inoculation of adenovirus-based vaccines, we generated several replication-defective adenoviruses expressing the glycoprotein D gene of pseudorabies virus under the control of four different promoters: major late promoter of adenovirus type 2, human cytomegalovirus immediate-early promoter/enhancer (CMV), Rous sarcoma virus-long terminal repeat promoter, and human desmin gene 5' regulatory region (DES). All the adenovirus constructs were able to fully protect mice, in the contrary of direct DNA inoculation of plasmids harboring the same transcription units. The far most effective adenovirus constructs, on the criterion of protective doses and specific antibody response induction, were those in which the foreign gene was driven by the DES or CMV promoter. Wide variations in promoter strength in vitro were evidenced in several cell culture types representative of putative target cells following muscular inoculation (myoblasts, myotubes, fibroblasts, macrophages, and endothelial cells). The level of efficacy in vivo, was not correlated with the level of expression in vitro in myotubes, but paralleled the level of expression in endothelial cells and in myoblasts. Together with previously published data, these results suggest that, following adenovirus injection, locally produced cytokines may induce myoblasts to act as local antigen presenting cells. PMID- 9400606 TI - Serial in vivo passage of HIV-1 infection in Macaca nemestrina. AB - In an earlier study we found that pigtailed macaques (Macaca nemestrina) that were experimentally infected with human immunodeficiency virus type 1 (HIV-1) initially became viremic and seroconverted, but HIV-1 replication diminished markedly over time. In an attempt to develop a longer term pathogenic model, blood from HIV-1-infected macaques was serially transfused into three groups of naive macaques. Transfer was successful through two transfusions as shown by repeated virus isolations and confirmed by the development of cell-free plasma viremia and by seroconversion. Three to five weeks after transfusion, plasma levels of HIV-1 RNA from several macaques in the first two groups exceeded those of the initially inoculated macaques. However, animals in the third group had diminished RNA levels, were virus culture negative, and did not seroconvert. Sequence analyses of env-region clones from infected animals revealed only minimal changes over the course of the passages. These results confirm HIV-1 replication in M. nemestrina during the acute phase of infection. However, adaptation of HIV-1 to a macaque-pathogenic variant did not occur during serial passage, possibly because the animals were able to restrict HIV-1 replication below a level required for a pathogenic variant to emerge. Whether such containment is a function of the host's immune response or a virus cell incompatibility remains to be determined. PMID- 9400607 TI - Epstein-Barr virus induces GM-CSF synthesis by monocytes: effect on EBV-induced IL-1 and IL-1 receptor antagonist production in neutrophils. AB - Neutrophils play an important role in the control of viral infections by releasing a variety of potent agents. We previously demonstrated that Epstein Barr virus (EBV) binds to human neutrophils and stimulates cytokine synthesis including interleukin-1 (IL-1) and IL-1 receptor antagonist (IL-1Ra). Since neutrophil functions are known to be modulated by the priming effect of granulocyte-macrophage colony-stimulating factor (GM-CSF), we therefore investigated the cellular source of GM-CSF synthesis following treatment of leukocytes with EBV and the effect of GM-CSF on the production of IL-1, IL-1Ra, and superoxide by EBV-treated neutrophils. In enriched-cell populations, only monocytes were found to produce GM-CSF in response to EBV, which was maximal after 12 h of incubation. The results obtained with UV-irradiated particles or EBV neutralized with monoclonal antibody 72A1 suggest that contact between the cell and the gp350 of the viral envelope is sufficient to induce the release of GM-CSF. On the other hand, GM-CSF differentially upregulated EBV-induced IL-1 and IL-1Ra production by neutrophils. Pretreatment of neutrophils with GM-CSF prior to EBV activation synergistically enhanced the production of IL-1 alpha and IL-1 beta, but only marginally affected IL-1Ra synthesis. In addition, GM-CSF was also found to synergistically enhance the superoxide production by neutrophils in response to EBV. Molecular analysis showed that GM-CSF did not alter the IL-1 beta and IL-1Ra mRNA synthesis induced by EBV, suggesting that GM-CSF could act at a posttranslational level. Local production of GM-CSF by monocytes in tissues invaded by EBV could serve to potentiate the host defense mechanisms directed toward the destruction of the infectious virus. PMID- 9400608 TI - Nucleotide sequence and genomic organization of Acyrthosiphon pisum virus. AB - The nucleotide sequence of the genomic RNA of Acyrthosiphon pisum virus was determined. The APV genome is 10,016 nucleotides in length, excluding the 3'-end poly(A) track, and contains two large open reading frames (ORFs), encoding proteins of 296,340 and 63,279 Da. The ORF1 is preceded by an untranslated leader sequence of 267 nucleotides. The ORF1 product contains sequence motifs characteristic of RNA-dependent RNA polymerases, chymotrypsin-like proteases, and helicases. Interviral sequence comparison revealed significant similarities with viruses belonging to the so-called picornavirus superfamily. The ORF2 is most likely expressed by a -1 translational frameshift and is followed by an untranslated sequence of 222 nucleotides. Internal amino acid sequences of three capsid proteins (66K, 34K, 23/24K) were determined. Comparison of the obtained amino acid sequences with the APV sequence disclosed that the structural proteins are located in the 3'-terminal half of the genome. The 34K protein is encoded by the ORF1, while the 66K protein contains both ORF1-(34K) and ORF2-derived sequences and is probably expressed by a translational frameshift. The 23/24K proteins most likely arise by proteolytic breakdown of the 34K protein. Although the deduced APV genomic organization in some aspects resembles that of the picornaviruses, its overall genomic organization indicates that APV is a distinct species only distantly related to the Picornaviridae. PMID- 9400609 TI - Heterogeneity and common features of defective hepatitis B virus genomes derived from spliced pregenomic RNA. AB - Defective hepatitis B virus (HBV) genomes derived from packaging and reverse transcription of spliced RNA pregenomes were reported to be associated with progression to chronic infection. Since only two types with similarly spliced regions were characterized so far we reasoned that additional "spliced" genome variants may exist. Therefore, we isolated a large number of defective HBV genomes from sera of seven chronic carriers by full-length PCR. Forty-eight were found to contain deletions caused by splicing as identified by cloning, subgenomic PCR, and sequencing. In total, 11 types of spliced genomes derived from excision of 10 different introns were present in various combinations in each serum. This diversity resulted from alternative usage of five splice donor and four acceptor sites present in most but not all HBV genotypes. All spliced genomes shared sequence elements essential for replication as well as for transcription of the pre-C and pregenome/C mRNAs and the X mRNA. Moreover, all contained the coding regions for the X protein and for precore/core or precore/ core fusion proteins but lacked the pre-S/S gene promoters. These data demonstrate substantial and HBV genotype-dependent diversity of spliced genomes from which a variety of aberrant precore/core fusion proteins and normal X protein but no functional envelope and P proteins could be expressed. These genomes and the encoded proteins may play a role in the viral life cycle, persistence, and pathogenesis. PMID- 9400610 TI - Negative regulation of a heterologous promoter by human cytomegalovirus immediate early protein IE2. AB - The HCMV IE2 protein promiscuously activates transcription of many viral and cellular genes. IE2 also negatively autoregulates its own expression by binding to a strategically positioned IE2 binding site, called CRS, located immediately downstream of the TATA box of the HCMV major IE promoter. Here we show that IE2 is able to repress transcription driven by a heterologous promoter, RSV LTR. Repression of RSV LTR by IE2 is completely dependent of DNA sequences downstream of the TATA box of RSV LTR. A DNA sequence, 5'-CGATACAATAAACG-3', evidently matching the proposed CRS consensus sequence, is located between nucleotides -13 and +1 (relative to the transcription start site) of RSV LTR. Three lines of evidence support the notion that this RSV CRS element is involved in the IE2 mediated repression of RSV LTR. First, introduction of mutation to the RSV CRS element renders to the mutant RSV LTR resistance to IE2-mediated repression. Second, a mutant IE2 defective in DNA binding cannot downregulate transcription from RSV LTR. Third, IE2 specifically binds to the wild-type, but not the mutant, RSV CRS element in vitro. These data, in conjunction with previous works, demonstrate that IE2 can passively repress transcription of homologous and heterologous promoters that contain a CRS element. PMID- 9400611 TI - Hantavirus pulmonary syndrome: CD8+ and CD4+ cytotoxic T lymphocytes to epitopes on Sin Nombre virus nucleocapsid protein isolated during acute illness. AB - In 1993 a number of cases of unexplained adult respiratory syndrome occurred in the southwestern United States. The illness was characterized by a prodrome of fever, myalgia, and other symptoms followed by the rapid onset of a capillary leak syndrome with hemoconcentration, thrombocytopenia, and pulmonary edema. Viral RNA sequences in the lungs identified a new member of the hantavirus genus, Sin Nombre virus (SNV), unique to North America. Pulmonary endothelial cells were heavily infected but were not necrotic. We speculated that this capillary leak syndrome was initiated by immune responses to the SNV-infected pulmonary endothelial cells. We isolated a CD8+ cytotoxic T lymphocyte (CTL) clone directly from the blood of a patient with the acute hantavirus pulmonary syndrome (HPS) which recognizes a SNV specific epitope on the virus nucleocapsid protein (aa 234 242) that is restricted by HLA C7 and produces IFN gamma but not IL-4. We identified a second CD8+ CTL epitope located within another site aa 131-139 on the nucleocapsid protein, which is HLA B35 restricted, and a CD4+ CTL epitope located on a third site on nucleocapsid protein aa 372-380 using lymphocytes obtained during HPS from another patient that were stimulated in vitro. Hantavirus specific CD8+ and CD4+ CTL may contribute to the immunopathology and capillary leak syndrome observed in the HPS. PMID- 9400612 TI - Analysis of a temperature-sensitive vaccinia virus mutant in the viral mRNA capping enzyme isolated by clustered charge-to-alanine mutagenesis and transient dominant selection. AB - We have previously reported the successful development of a targeted genetic method for the creation of temperature-sensitive vaccinia virus mutants [D. E. Hassett and R. C. Condit (1994) Proc. Natl. Acad. Sci. USA 91, 4554-4558]. This method has now been applied to the large subunit of the multifunctional vaccinia virus capping enzyme, encoded by gene D1R. Ten clustered charge-to-alanine mutations were created in a cloned copy of D1R. Four of these mutations were successfully transferred into the viral genome using transient dominant selection, and each of these four mutations yielded viruses with plaque phenotypes different from that of wild-type virus. Two of the mutant viruses, 516 and 793, were temperature sensitive in a plaque assay. Mutant 793 was also temperature sensitive in a one-step growth experiment. Phenotypic characterization of the 793 virus under both permissive and nonpermissive conditions revealed nearly normal patterns of viral protein and mRNA synthesis. Under nonpermissive conditions the 793 virus was defective in telomere resolution and blocked at an intermediate stage of viral morphogenesis. In vitro assays of various capping enzyme activities revealed that in permeabilized virions, enzyme guanylylate intermediate formation was reduced and methyltransferase activity was thermolabile, while in solubilized virion extracts enzyme guanylylate activity was reduced and both guanylyltransferase and methyltransferase activities were absent. Thus, the 793 mutation affects at least two separate enzymatic activities of the capping enzyme, guanylyltransferase and methyltransferase, and when incorporated into the virus genome, the mutation yields a virus that is temperature sensitive for growth, telomere resolution, and virion morphogenesis. PMID- 9400613 TI - Interaction of the human protein kinase PKR with the mouse PKR homolog occurs via the N-terminal region of PKR and does not inactivate autophosphorylation activity of mouse PKR. AB - The RNA-dependent protein kinase (PKR) is implicated in the antiviral and antiproliferative actions of interferon. Mutant forms of human PKR display a transdominant behavior when expressed in transfected cells. The potential for the human PKR protein to physically interact with the mouse PKR homolog has therefore been examined. The yeast two-hybrid system was used to probe the association between mouse and human PKR proteins as measured by activation of two Gal4 responsive reporter genes, HIS3 and IacZ. Expression of full-length wild-type mouse PKR(1-515)WT as a Gal4 fusion protein did not exhibit the growth suppression phenotype in yeast characteristic of wild-type human PKR(1-551)WT. Coexpression of mouse PKR(1-515)WT as a Gal4 DNA-binding domain fusion with either the catalytic-deficient human PKR(1-551) K296R mutant, the RNA-binding deficient human PKR(1-551)K64E/K296R double mutant, or wild-type mouse PKR(1 515)WT as full-length PKR-Gal4 activation domain fusions resulted in activation of the HIS3 and lacZ reporters. The N-terminal RNA-binding region of human PKR, both WT and the K64E RNA-binding-deficient mutant, also interacted with mouse PKR(1-515)WT sufficiently to activate the reporters but the human catalytic region did not. Mouse and human full-length PKR proteins expressed as glutathione S-transferase (GST) fusions in Escherichia coli were purified on Sepharose beads. Using GST-PKR fusion chromatography, direct physical interaction between the mouse and human PKR homologs was established. Intraspecies PKR interactions were more efficient than interspecies PKR interactions, and interactions between RNA binding-sufficient PKR proteins were more efficient than those involving an RNA binding mutant as measured by binding to GST-PKR protein Sepharose beads. The N terminal region of human PKR within amino acids 1-184 was sufficient for binding mouse PKR. Purified mouse full-length PKR(1-515)WT GST fusion protein retained kinase activity on Sepharose beads, but the activity was not impaired by association with either the full-length or the N-terminal region of human PKR. PMID- 9400614 TI - A point mutation in the Sendai virus accessory C proteins attenuates virulence for mice, but not virus growth in cell culture. AB - A mutant Sendai virus (SevMVC), which grows much better than its progenitor virus (SeVM) in cell culture, but, in strong contrast to SeVM, is totally avirulent for mice, has been described. SeVMVC contains two amino acid substitutions relative to SeVM, namely, F170S in the C protein and E2050A in the L protein. We have examined which substitutions were responsible for the above phenotypes by exchanging the C gene of our reference strain Z with those of SeVH (another reference strain), SeVM, and SeVMVC, in turn. We have found that the F170S mutation in the CMVC protein is responsible both for enhanced replication in cell culture and for avirulence in mice. Avirulence appeared to be due to restricted viral replication primarily after day 1, implicating some aspect of innate immunity in this process. The SeV C proteins thus appear to be required for multiple cycles of replication in mice. PMID- 9400615 TI - Increasing the ratio of PP2A core enzyme to holoenzyme inhibits Tat-stimulated HIV-1 transcription and virus production. AB - We demonstrated previously that PP2A exists in many cell types as two abundant forms: (1) holoenzyme composed of two regulatory subunits, A and B, and a catalytic subunit C; and (2) core enzyme consisting of the A and C subunits. These two forms have different substrate specificities. Since published data suggested that HIV-1 transcription may be regulated by a cellular protein phosphatase, it was of interest to determine whether changing the ratio between PP2A core and holoenzyme affects HIV-1 gene expression. This question was addressed by expression in COS cells of an N-terminal mutant of the A subunit, A delta 5, which binds the C but not the B subunit. This resulted in an increase in the amount of core enzyme and a decrease in the amount of holoenzyme concomitant with the expected change in phosphatase activity. Tat-stimulated transcription from the HIV-1 LTR was inhibited 5-fold by mutant A delta 5, whereas mRNA synthesis directed by the actin promoter was not affected. Furthermore, virus production in COS, HeLa, and Jurkat T cells was inhibited 45-, 5-, and 3-fold, respectively, by mutant A delta 5. These results demonstrate that the balance between PP2A holoenzyme and core enzyme is important for HIV-1 gene expression and virus production. PMID- 9400616 TI - Construction and characterization of vaccinia direct ligation vectors. AB - Poxvirus vectors are extensively used as expression vehicles for protein and antigen expression in eukaryotic cells. Customarily, the foreign DNA is introduced into the poxvirus genome by homologous recombination. An alternative method using direct ligation vectors has been used to efficiently construct chimeric genomes in situations not readily amenable for homologous recombination. We describe the construction and characterization of a new set of direct ligation vectors designed to be universally applicable for the generation of chimeric vaccinia genomes. These vectors contain the pair of unique restriction sites NotI and ApaI to eliminate religation of poxvirus arms and fix the orientation of the insert DNA behind strongly expressing constitutive vaccinia promoters. The insertion cassette has been placed at the beginning of the thymidine kinase gene in vaccinia to use drug selection in the isolation of recombinants. These viruses provide a set of universally applicable direct ligation poxvirus cloning vectors, extending the utility of poxvirus vectors for construction and expression of complex libraries. PMID- 9400617 TI - Molecular studies on bromovirus capsid protein. IV. Coat protein exchanges between brome mosaic and cowpea chlorotic mottle viruses exhibit neutral effects in heterologous hosts. AB - Two members of the bromovirus group, brome mosaic virus (BMV) and cowpea chlorotic mottle virus (CCMV), selectively infect barley and cowpea, respectively, and also differ in their ability to systemically infect a common permissive host, Chenopodium quinoa. CCMV is confined to inoculated leaves of C. quinoa, whereas BMV causes rapid systemic mottling. To examine whether host specific determinants for systemic movement of BMV and CCMV in each of these hosts are localized in the coat protein (CP), sequences encoding this gene were exchanged between biologically active clones of BMV RNA3 (B3) and CCMV RNA3 (C3) to create chimera expressing heterologous CP genes (B3/CCP and C3/BCP). Inoculation of each chimera with its respective wild-type (wt) RNAs 1 and 2 to barley or cowpea or C. quinoa plants resulted in symptom phenotype and long distance movement characteristics similar to those of the parental virus donating RNAs 1 and 2. These observations suggest that neither BMV CP nor CCMV CP has host specific determinants for long distance movement. Inoculation of additional recombinant viruses, constructed by reassorting wt genomic RNAs 1 and 2 of BMV and CCMV with either heterologous wt RNA3 (i.e., B1 + B2 + C3 and C1 + C2 + B3) or heterologous chimeric RNA3 (i.e., B1 + B2 + C3/BCP and C1 + C2 + B3/CCP), to susceptible hosts resulted only in localized infections. The significance of these observations in relation to bromovirus movement is discussed. PMID- 9400618 TI - The RNA binding region of the paramyxovirus SV5 V and P proteins. AB - The V/P gene of simian virus 5 (SV5) encodes two proteins, V (222 residues) and the phosphoprotein P (392 residues). The V and P proteins are amino coterminal for 164 residues, but they have unique carboxy termini due to addition of two nontemplated G residues to the P mRNA during transcription. We have shown that the V and P proteins bind RNA by using both Northwestern blot analysis and ultraviolet-light crosslinking. The RNA-binding region has been mapped to a region in the P and V proteins which contains five basic residues (K74, K76, K77, R79, K81). Either deletion of the basic residues or substitution of the basic residues with alanine inhibited RNA binding by the V or P proteins. PMID- 9400619 TI - Analysis in vivo of turnip crinkle virus satellite RNA C variants with mutations in the 3'-terminal minus-strand promoter. AB - Turnip crinkle virus and its associated RNA, sat-RNA C, share similar, but not identical hairpins near their 3' ends and terminate with CCUGCCC-OH, which forms a single-stranded tail. With an in vitro transcription system containing partially purified TCV RdRp, the 3'-terminal 29 bases making up the hairpin and single-stranded tail were previously demonstrated to be required for transcription, and alterations in the stem, but not the loop, could affect template activity (C. Song and A. E. Simon, 1995, J. Mol. Biol. 254, 6-14). We have now analyzed sat-RNA C mutants in the 3' hairpin for ability to accumulate in vivo. While active templates in vitro were able to accumulate in vivo, some very weak templates in vitro were also able to accumulate in vivo without reversion or second-site alterations. Computer models of hairpin structure indicated that biologically active promoters could have hairpins less stable than wild type, with loops of variable length and sequence, and without a need for a 6 base single-stranded tail. In addition, transcripts containing compensatory exchanges in the upper stem region that had limited activity in vitro were biologically active in vivo, indicating that positioning of specific bases in the stem is not required to produce an active minus-strand promoter. PMID- 9400620 TI - The coat protein of turnip crinkle virus is involved in subviral RNA-mediated symptom modulation and accumulation. AB - Some satellite (sat-) and defective interfering (DI) RNAs associated with plant viruses intensify or ameliorate the symptoms of the virus. We recently demonstrated that the TCV coat protein (CP) is involved in symptom modulation by sat-RNA C. Two additional subviral RNAs have now been tested for effect of the CP on symptom modulation. DI RNA G, which normally intensifies the symptoms of TCV, is able to attenuate symptoms if the TCV CP is replaced with the CP of cardamine chlorotic fleck virus. DI RNA G had no effect on the symptoms of TCV with a single base alteration in the CP open reading frame, unlike sat-RNA C, which was able to ameliorate the symptoms of the mutant TCV. Using a hybrid sat-RNA constructed from sat-RNA C and TCV (which shares a similar 3'-end region with DI RNA G), the 3'-terminal 53 bases of sat-RNA C were found to be involved in symptom attenuation, which was directly correlated with the lack of detectable viral genomic RNA in whole plants. Sat-RNA D had no effect on the symptoms of mutant or wild-type TCV. The accumulation of TCV subviral RNAs in plants and protoplasts was also found to be strongly influenced by the presence or absence of the wild-type TCV CP. PMID- 9400621 TI - Sialyl-Lewis(x) sequence 6-O-sulfated at N-acetylglucosamine rather than at galactose is the preferred ligand for L-selectin and de-N-acetylation of the sialic acid enhances the binding strength. AB - Oligosaccharide sequences based on sialyl-Lewis(x) with 6-O-sulfation at galactose (6'-sulfo) or at N-acetylglucosamine (6-sulfo) and expressed on high endothelial venules are considered likely endogenous ligands for the leukocyte adhesion molecule, L-selectin. In the course of high performance TLC of three hexaglycosylceramides 6'-sulfo sialyl Lewis(x), 6-sulfo sialyl Lewis(x), and 6',6 bis-sulfo sialyl Lewis(x), synthesized chemically for selectin recognition studies, two minor byproducts were detected and isolated from each parent compound. By liquid secondary ion mass spectrometry these were identified as isomers containing a de-N-acetylated sialic acid or having a modified carboxyl group. Binding experiments with the parent compounds and the non-sulfated sialyl Lewis(x) glycolipid show that 6-sulfation potentiates, whereas 6'-sulfation virtually abolishes L-selectin binding. Thus the hierarchy of binding strengths were 6-sulfo sialyl > sialyl = 6',6-bis-sulfo sialyl >> 6'-sulfo sialyl Lewis(x). Whereas modification of the sialic acid carboxyl group markedly impaired L selectin binding, de-N-acetylation resulted in enhanced binding. The natural occurrence on high endothelial venules of this 'super-active' de-N-acetylated form of 6-sulfo sialyl Lewis(x), and related structures, now deserves investigation. PMID- 9400622 TI - Detection of molecular determinants and epitope mapping using MALDI-TOF mass spectrometry. PMID- 9400623 TI - Cytokine mRNA profiles during the course of experimental Haemophilus influenzae bacterial meningitis. AB - Intraperitoneal inoculation of Haemophilus influenzae type b (Hib) to 3-week-old Sprague-Dawley rats resulted in nonlethal meningitis with high levels of leukocytes in the cerebrospinal fluid (CSF) and positive bacterial culture. Using in situ hybridization, levels of cytokine mRNA-expressing cells were determined in the brain, CSF, and spleen from Hib-inoculated and uninfected control rats. IFN-gamma, IL-1 beta, IL-4, IL-6, IL-10, IL-12, and TNF-alpha mRNA levels were elevated at 12 hr postinoculation (pi) in spleen and CSF. At this time point, strong expression of IL-6 and TGF-beta was detected in the brain, and also of IL 10 at 48 hr while IFN-gamma and IL-12 were expressed at very low levels throughout the observation time. Delayed cytokine induction occurred in CSF compared to spleen and brain. TGF-beta was high in CSF at 48 hr, and some elevation of IL-1 beta, IL-6, IL-10, TNF-alpha, IFN-gamma, and IL-12 was evident at 72 hr pi. This may suggest measures that promote production of TGF-beta and/or IL-10 should be evaluated in treatment of bacterial meningitis. PMID- 9400624 TI - Synchronous decline of serum-soluble HLA class I antigen and beta-cell function in insulin-dependent diabetes mellitus. AB - Serum-soluble HLA class I molecules (sHLA) have immunomodulatory functions and their serum levels correlate with the HLA class I phenotype. We studied longitudinal changes of serum sHLA levels in insulin-dependent diabetes mellitus (IDDM). A total of 198 serum samples were obtained from 40 IDDM patients before and after IDDM onset. sHLA was assayed by a sandwich ELISA. sHLA levels in IDDM patients at the initiation of insulin therapy (IDDM onset) were markedly reduced compared with those in normal controls (334.2 +/- 26.3 ng/ml vs 492.4 +/- 55.5 ng/ml, mean +/- SEM, P = 0.0038). They fell sharply during 6 months before and after the onset of IDDM. The dynamic profile of sHLA and the time course of beta cell loss were different between IDDM patients with and without HLA-A24. In those with HLA-A24, sHLA became significantly lower than normal controls with HLA-A24 at IDDM onset. Recovery of their sHLA values occurred at 3 years from IDDM onset. On the other hand, in those without HLA-A24, sHLA levels began to decrease since the onset of IDDM and became significantly lower than normal controls without HLA A24 at 4 years after the onset. Recovery of sHLA occurred at more than 6 years from the onset. An early (within 18 months), complete loss of beta-cell function occurred in 5 of 13 IDDM patients with HLA-A24 compared with 1 of 14 of those without HLA-A24 (P = 0.077). A late (more than 36 months after the onset of IDDM), complete loss of beta-cell function occurred in 7 of 14 IDDM patients without HLA-A24 but in none of 13 of those with HLA-A24 (P = 0.0058). These results indicate that the decline of sHLA is synchronous with massive beta-cell destruction, and that these events occur during a short period in IDDM patients with HLA-A24, whereas they occur during a relatively long period in those without HLA-A24. PMID- 9400625 TI - Evaluating the role of Th0 and Th1 clones in autoimmune thyroid disease by use of Hu-SCID chimeras. AB - To study the role of Th0 and Th1 cells in autoimmune thyroid disease, thyroid tissues from patients with Graves' disease (GD), Hashimoto's thyroiditis (HT), and colloid nodular disease were xenografted into SCID mice, followed by ip injection of peripheral blood mononuclear cells (PBMC), T cell lines, and T cell clones (TCC). The antigen-specific TCC reactive to TSH receptor (TSH-R), thyroid peroxidase (TPO), or thyroglobulin (Tg), and their respective peptides, were classified into Th0 (secreting IL-4 and/or IL-5 and IFN-gamma) and Th1 (secreting IFN-gamma) according to their cytokine profile. Engraftment of autologous or HLA matched allogeneic CD4+ thyroid-specific clones with Th0 or Th1 phenotypes induced the production of total IgG and thyroid-specific autoantibodies by B cells present in xenografted thyroid tissues. TSH-R-specific clones mainly enhanced thyroid-stimulating antibodies (TSAb) production, while clones reactive to TPO and Tg increased the synthesis of TPO and Tg autoantibodies. Total IgG production, but not TSAb, was also stimulated by PBMC and TSH-R lines. TSAb correlated with the viability and hyperplasia of thyroid follicles, but not with the serum T3 levels, which were normal. Thyroid tissue viability was maintained or increased by antigen-specific Th0 clones, and decreased by Th1 clones reactive to TSH-R or TPO. Thyroid lymphocytic infiltration was variable; however, Th0 and Th1 clones from HT patients caused high degree of lymphocytic infiltration compared to the control groups. These results demonstrate for the first time that T cells clones reactive to specific epitopes of TSH-R, TPO, or Tg can generate antibody-mediated and/or cell-mediated responses in the xenografted thyroid tissue microenvironment. Such effects depend on clonal specificity, HLA class II restriction, and cytokine profile of the clone. Th0 clones reactive to TSH-R stimulate both total IgG production and TSAb in SCID mice engrafted with thyroid tissue from GD patients. Th0 and Th1 clones specific for TPO and Tg also function as helper T cells, stimulating total IgG synthesis and autoantibodies against TPO and Tg. Th1 clones may also cause tissue destruction in GD and HT. PMID- 9400626 TI - Dinitrophenyl-modified autologous melanoma vaccine induces a T cell response to hapten-modified, melanoma peptides. AB - Active specific immunotherapy with dinitrophenyl (DNP)-modified autologous melanoma vaccine elicits inflammatory responses in metastatic tumor sites. Postsurgical adjuvant immunotherapy with this vaccine prolongs survival in stage III melanoma patients. We have reported that, after administration of DNP modified melanoma vaccine, T cell responses to DNP-modified autologous tumor cells are demonstrable in vivo and in vitro. These responses are hapten specific and MHC restricted. To elucidate this phenomenon, we investigated the immune response to DNP-modified peptides eluted from autologous cells. Short peptides were extracted from DNP-modified and unmodified autologous melanoma cells by an acid elution technique and HPLC fractionation. Peptides were also extracted from DNP-modified and unmodified, EB virus-transformed, autologous B lymphoblasts. These various peptide fractions were loaded onto autologous B lymphoblasts and tested for ability to elicit a response by a DNP-specific T cell line as measured by IFN-gamma production. Unexpectedly, stimulatory activity of peptides from DNP modified melanoma cells was confined to a single HPLC fraction. Spectrometric analysis of this fraction confirmed modification of peptides with DNP. A weaker T cell response was observed to a single HPLC fraction of DNP-modified peptides from the patient's B lymphoblasts. No T cell response was elicited by corresponding fractions of peptides eluted from unmodified melanoma cells or B lymphoblasts. These findings demonstrate the human T cell response to DNP modified autologous melanoma cells is mediated by hapten-modified, MHC-associated peptides. Further investigation of these peptides could lead to a new strategy for peptide-based cancer immunotherapy. PMID- 9400627 TI - Lymphocyte phenotyping in infants: maturation of lymphocyte subpopulations and the effects of HIV infection. AB - Changes in the distribution of lymphocyte subpopulations in infants with perinatally acquired HIV infection are confounded by the rapid changes that are the result of normal maturation of the immune system. We describe the changes in seven lymphocyte phenotypes (CD3+ CD4+, CD3+ CD8+, CD8+ HLA- DR+, CD8+ CD38+, CD8+ CD57+, CD3-/ CD16+ 56+, and CD19+) over the first 2 years of life in 390 HIV 1 exposed but uninfected and 98 HIV-1-infected infants enrolled in the Women and Infants Transmission Study. The greatest changes in uninfected infants were declines in the CD3+ CD4+ lymphocytes and increases in CD8+ HLA- DR+ and CD19+ lymphocytes. All phenotypes were affected by HIV infection but the greatest changes were declines in the CD3+ CD4+ subset and increases in the CD3+ CD8+ and CD8+ HLA- DR+ subsets. Thus, this study provides reference data for the maturational changes in lymphocyte phenotypes in HIV-exposed but uninfected infants and describes the overall changes that occur with perinatally acquired HIV infection. PMID- 9400628 TI - Role of T-cell subsets in acute and persistent E-55+ murine leukemia virus infection in susceptible progressor and resistant long-term nonprogressor mouse strains. Women and Infants Transmission Study. AB - Previous studies from this laboratory have demonstrated that E-55+MuLV-infected BALB/c-H-2k (BALB.K) mice progress to develop thymic lymphoma about 7 months after infection whereas infected C57BL/10-H-2k (B10.BR) mice are long-term nonprogressors that fail to develop disease even after 2 years of infection. Both resistant long-term nonprogressor (B10.BR) and progressor (BALB.K) mice generate an early immune response that results in a dramatic decrease in the number of virus-infected cells. Despite this early immune response, mice from both strains become persistently infected. However, resistant B10.BR mice also demonstrate a late T-cell-mediated response that may be causally related to long-term nonprogression whereas susceptible BALB.K mice fail to demonstrate this late T cell response. In the present studies, the T-cell subsets involved in the effective early immune response in both B10.BR and BALB.K mice as well as the late T-cell response in B10.BR mice were determined by in vivo antibody-mediated depletion. Results from these studies demonstrate that during the early acute phase of infection, elimination of CD4+ T cells ablated the ability of both BALB.K and B10.BR mice to decrease the burden of virus-infected cells. However, elimination of CD8+ T cells ablated this result in BALB.K but not B10.BR mice. Thus, despite the fact that both immunocompetent B10.BR and BALB.K mice are able to decrease the number of virus-infected cells during the early acute phase of infection, there is a difference in the T-cell subsets that mediate this effect in these strains of mice. In addition, characterization of the late immune response that keeps virus at very low levels during the persistent stage of virus infection in resistant B10.BR mice demonstrated that simultaneous elimination of both CD4+ and CD8+ T cells allowed the emergence of virus-infected cells whereas the elimination of either subset alone showed no effect compared to untreated control mice that are immunologically intact. Since B10.BR and BALB.K are identical with respect to their H-2k-haplotypes, it appears that the differences between these strains with respect to the generation of effective early and late anti-virus immune responses are regulated by a non-H-2-linked gene(s). PMID- 9400629 TI - Anti-gliadin antibodies in patients with celiac disease cross-react with enterocytes and human calreticulin. AB - One of the characteristic features of celiac disease is an increase in anti gliadin antibodies (Abs). Recently we found that some of the monoclonal Abs to gliadin cross-react with molecules on rat enterocytes. One of these cross reacting molecules was identified as rat calreticulin. This study shows that the levels of serum IgA Abs to gliadin, rat, and human enterocytes; purified enterocyte antigens; and calreticulin in sera from patients with active disease were significantly higher than in patients on a gluten-free diet and healthy controls (P < 0.001). Anti-gliadin Abs were isolated by affinity chromatography from the sera of six active celiac patients. The reactivity of these anti-gliadin Abs was demonstrated to be significantly higher (P < 0.05) with human enterocytes and human calreticulin than with other antigens tested. Furthermore, using isolated patients' anti-gliadin Abs bound to Sepharose 2B, two main proteins of molecular mass 62 and 66 kDa were purified from a lysate of human enterocytes. The 62-kDa enterocyte antigen was identified as human calreticulin. These findings suggest that anti-gliadin Abs may play a pathogenic role in celiac disease by cross-reacting with enterocytes. Calreticulin in enterocytes may be one of the putative targets for autoimmune reactions. PMID- 9400630 TI - Interactions of lymphocytes from patients with psoriatic arthritis or healthy controls and cultured endothelial cells. AB - Psoriatic arthritis (PA) is an inflammatory rheumatic disease that can concomitantly occur in patients with psoriasis vulgaris. Psoriatic synovitis shows alterations of the synovial microvasculature. Inflammatory cells adhere to endothelial cells (EC) and migrate through the vascular wall of postcapillary venules located in the subintimal layer of the synovial membrane. The aim of our study was to investigate, first, the phenotype of lymphocytes (LC) of PA patients using flow cytometry (FC) with regard to activation antigens and adhesion molecules; second, the adhesion of LC of PA patients on cultivated resting or activated (with thrombin, LPS, IFN-gamma, or TNF-alpha) human umbilical vein endothelial cells (HUVEC) by counting the Feulgen-stained nuclei of both adherent LC and HUVEC using image analysis; and third, the synthesis of IL-6 and IL-8 in both LC and HUVEC 24 hr after cell contact. These cytokines were determined qualitatively by immunofluorescence and quantitatively at the single-cell level by FC as well as in the supernatants of the cultures using commercial cytokine ELISAs. Fourth, we investigated whether or not the LC adhesion on HUVEC as well as the cytokine production could be inhibited by monoclonal antibodies against LC or EC-specific adhesion molecules. In contrast to controls PA patients showed an increased surface expression of CD11a, b, and c as well as of CD44 but a reduced surface expression of CD49d/CD29, and CD49e/CD29, and cell-bound fibronectin on CD3+ LC. The activation markers CD25 and HLA-DR were found to be slightly enhanced in PA. The cell adhesion was generally enhanced in PA patients vs controls. It could be reduced with monoclonal antibodies (MoAbs) against CD11a and CD18 on IFN-gamma- or TNF-alpha-activated HUVEC but was generally enhanced after treatment of HUVEC with MoAbs against CD54, CD62E, or CD106. Due to LC adhesion on HUVEC IL-6 and IL-8 were produced in significantly higher amounts in PA patients compared to controls. This effect occurred already in resting but was enhanced in activated HUVEC. While IL-6 is mainly produced by HUVEC but also in smaller quantities by LC, IL-8 is synthesized only by HUVEC and could be modified by preincubation with MoAbs against LC- or EC-specific adhesion molecules in parallel to the cell adhesion. The experiments show that the main adhesion pathway in LC homing of PA patients is the interaction of the LC adhesion molecule CD11a/CD18 with CD54 on EC followed by an enhanced synthesis of proinflammatory and chemotactic cytokines. These results favor the hypothesis that the pathological alterations of the microvasculature in PA patients are generated by altered homing processes. PMID- 9400631 TI - Adoptively transferred EAE in mice bearing the lpr mutation. AB - We have recently developed approaches for the generation of encephalitogenic T cell clones from mouse strains considered resistant to experimental allergic encephalomyelitis (EAE). By allowing for the direct use of knockout and mutant strains of mice, such clones allow for the efficient characterization of the relevance of specific gene products in the effector phase of EAE. Recent studies have suggested that Fas/FasL-mediated cell death may play a role in the pathogenesis of MS. To assess the role of Fas/FasL in EAE, we have tested the ability of wild-type C57BL/6-derived, encephalitogenic T cell clones to mediate adoptively transferred EAE in Fas-deficient C57BL/6-lpr mice. We now report that mice with the lpr mutation are fully susceptible to the adoptive transfer of EAE. Our results suggest that Fas/FasL-mediated cell death in the central nervous system does not play an integral role in the effector phase of acute EAE. PMID- 9400632 TI - Patterns of in vitro anti-human immunodeficiency virus type 1 antibody production in long-term nonprogressors. AB - With the aim of evaluating the specific pattern of in vitro antibody production (IVAP) in human immunodeficiency virus type 1 (HIV-1)-infected long-term non progressors (LTNPs), we tested 20 subjects who had remained asymptomatic for more than 8 years with a CD4+ cell count higher than 500/microliter and 59 patients at different stages of HIV-1 infection as controls. In cell cultures, IVAP was detected in 14 out of 20 LTNPs (70%), in 5 out of 6 recent seroconverters (83%), and in all the other control patients. Anti-p24 antibody production was significantly lower in LTNPs than in asymptomatic patients with a more recent infection. Recent seroconverters and patients with AIDS did not produce anti-p24 antibodies (P = 0.02). Anti-gp160 antibodies were produced by peripheral blood mononuclear cells from LTNPs in 12/20 cases. CD4+ cell count was significantly higher in IVAP-negative than in IVAP-positive LTNPs (P = 0.013), while the viral load was not significantly different. Specific anti-HIV-1 antibody production did not seem to be a correlate of long-term nonprogression. PMID- 9400633 TI - The use of clinical practice guidelines (CPGs) to evaluate practice and control costs in ventriculoperitoneal shunt management. AB - BACKGROUND: As a step toward maximizing the quality and cost-effectiveness of neurosurgical care, we designed clinical practice guidelines (CPGs) for the management of VP shunt malfunctions and infections at a tertiary care pediatric teaching institution. The detailed CPGs determine the use of radiographic studies, laboratory tests, and invasive procedures in the management of this problem. One purpose of the CPGs is to provide clear clinical guidelines for the medical trainee, thereby reducing variability in care and unnecessary utilization of resources. METHODS: The CPGs were developed in stages over a 2-year period. The practice patterns in our institution for the management of shunt malfunctions and infections were articulated. They were compared with those published in the neurosurgical literature, and areas of clinical decision-making variability were identified. Preliminary guidelines were formulated, and data regarding patient care were prospectively collected. Based on this data, final CPGs were formulated and implemented. Total and itemized hospital charges for patients managed according to the CPGs were compared with those for patients in the 3 years before CPG implementation. RESULTS: CPG-managed patients had generally lower total and itemized charges as compared with control patients. Decreased charges per hospital day and charges for shunt films in the CPG group were statistically significant. CONCLUSIONS: The process by which the CPGs were developed and implemented, as well as the CPGs themselves, are described. We also present the clinical, demographic, and financial data that were prospectively collected for all patients managed within the CPGs over an initial 1-year period and compare it with data obtained for control groups of shunt malfunction patients admitted during the 3 years before implementation of the CPGs. We find a trend toward reduction of charges after implementation of the CPG. PMID- 9400634 TI - Stereotaxy reduces cost of brain tumor resection. AB - BACKGROUND: Health care professionals are under increasing pressure to contain the cost of health care. Simultaneously, medical technology continues to advance. Medical institutions must therefore consider the costs and benefits before using a new technology. Using a direct costing system, we determined the cost efficacy of stereotaxy applied to the resection of brain mass lesions. METHODS: Twenty nine patients underwent a stereotactically guided craniotomy and brain tumor resection. Fifteen of them underwent general and fourteen received local anesthesia. Twelve other patients, comprising a historical reference group, underwent a standard craniotomy and brain tumor resection under general anesthesia. costs were determined for every hospital charge item in all patients. Cost efficiency was then compared between the two groups. RESULTS: Patients treated stereotactically incurred additional costs in frame placement and neuroimaging. These costs were offset by savings in operating room time, patient acuity, length of stay, respiratory care, and medications. Savings were greatest for patients who had local anesthesia. Overall, patients treated by stereotactic craniotomy had a total hospitalization cost of $8,495.19, whereas those treated with standard craniotomy incurred a cost of $11,365.23 (p < 0.001). CONCLUSION: Stereotaxy is cost effective for the surgical treatment of brain tumors. Accurate estimates of cost can justify the use of medical technology. Directly measured cost data is a useful index for any cost containment program. PMID- 9400635 TI - Lumbar disc surgery in a fixed compensation population: a model for influence of secondary gain on surgical outcome. AB - BACKGROUND: Reported outcomes in patients undergoing surgical procedures for lumbar disc herniation are poorer in patients eligible for workers' compensation or with pending litigation. In the civilian community, the amount of compensation for one's disability is variable and thus its influence on surgical outcome is difficult to quantify. In the military, all members are covered by a standardized workers' compensation system, and have generally standardized work requirements, a standard pay scale, and third party evaluation of disability based on the Veterans Affairs rating system. This made the military a good system in which to study the effect of potential compensation on surgical outcome. METHODS: The study population consisted of active duty military members who underwent sequential lumbar microdiscectomies over a 31-month period. Omitted were lumbar fusions, decompressive laminectomies, and far lateral discectomies. Clinical and demographic variables, along with financial data for each patient were derived from these data. A good result was defined as return to active military duty. RESULTS: Three hundred forty-nine lumbar discectomies were performed in 348 active duty military members. Overall, 75.3% (262) of the 348 patients were able to return to full military duty after surgery, and 24.7% (86) received disability compensation. Chi-square univariate analysis showed higher compensation incentive was a significant determinant of poor surgical outcome (p = 0.0021). The influence of compensation incentive was proportional to the amount of anticipated payout, and relative to a military service member's usual income. In mutivariate analysis, lower base pay (0.0005) and female gender (p = 0.038) were predictive of poor outcome. CONCLUSIONS: Secondary gain in the form of disability pay has a proportionally adverse effect on outcome following lumbar disc surgery. Although studying this issue in the military system allowed standardization of secondary gain values, the influence of other factors could not be eliminated entirely. Potential disability pay is proportionally greater in lower ranked service members. Thus, other variables such as income level, education, and job satisfaction may contribute to the poorer results in this subgroup of military members. PMID- 9400636 TI - Cost advantages of two-level anterior cervical fusion with rigid internal fixation for radiculopathy and degenerative disease. AB - BACKGROUND: Conventional anterior cervical discectomy with fusion is thought to require postoperative neck immobilization for the promotion of bony fusion. Rigid internal fixation with anterior cervical plates may decrease graft-related complications and provide immediate stability. This stability may obviate postoperative external immobilization. METHODS: This report reviews one surgeon's experience with the use of rigid internal fixation for two-level anterior cervical discectomy and fusion for radiculopathy to promote early mobilization without external bracing. It compares outcomes and costs with a similar population of patients treated with anterior cervical discectomy and fusion who did not undergo rigid internal fixation. We compared patients who underwent two level allograft anterior cervical discectomy and fusion with or without rigid internal fixation between 1989 and 1994 performed by a single surgeon (FJP) to evaluate the cost advantages and outcome of each procedure. All patients had clinical evidence of cervical radiculopathy unresponsive to medical therapy with magnetic resonance imaging confirmation of the appropriate nerve root impingement. Thirty-nine patients underwent two-level Cloward allograft fusion using Synthes anterior cervical locking plates, 25 underwent identical fusion without plating. Follow-up was 6 months to 4 years (mean, 31 months). RESULTS: Twenty-three of 25 patients in the nonplated group and 36 of 39 patients in the plated group achieved excellent or good outcomes using the Odom criteria. There were six complications (two major and four minor) in each group. Patients who underwent plating returned to light activities (mean, 17 vs. 29 days), driving (28 vs. 57 days), and unrestricted work (66 vs. 136 days) sooner than non-plated patients (p < 0.05, paired t test). No patient with plates was given external immobilization. CONCLUSIONS: Two-level anterior cervical discectomy and fusion with anterior plating for radiculopathy is safe, effective, and seems to provide shorter convalescence compared with conventional anterior cervical discectomy and fusion. Patients returned to unrestricted work sooner, thus reducing short-term disability. Rigid internal fixation may provide cost advantages to patients and insurance disability providers. The authors conclude that the increased cost of treatment for rigid internal fixation is more than offset by the benefits of earlier mobilization. PMID- 9400637 TI - Spinal instrumentation with a low complication rate. AB - BACKGROUND: Spinal instrumentation has become an increasing part of the armamentarium of neurosurgery and neurosurgical training. For noncontroversial indications for spine fusion the arthrodesis rate seems to be better. For both noncontroversial and controversial indications, the reported complication rate with spinal instrumentation tends to be greater than that with noninstrumented spine surgeries. These reported complications include a 2-3% neurologic injury rate, 3-45% reoperation rate for implant failure, and inflection rates of 5-10%. Therefore, we report on 299 cases that have undergone spinal instrumentation placed exclusively by neurosurgeons with a very low complication rate. METHODS: Two hundred ninety-nine consecutive spinal instrumentation cases performed exclusively by neurosurgeons at Indiana University Medical Center were analyzed for complications related to spinal instrumentation. The spinal instrumentation placed consisted of 195 anterior cervical locking plates, 22 cases of posterior cervical instrumentation, 9 cases of combined anterior locking plates with posterior cervical instrumentation, 14 anterior thoracolumbar plates, 51 posterior thoraco-lumbar instrumentation cases, and 8 combined anterior/posterior thoracolumbar instrumentation cases. RESULTS: The mean follow-up is 40 months (6 95). There was one perioperative death unrelated to the spinal instrumentation. There were no neurologic injuries and there has been no hardware infection to date. There were two dural tears, three superficial wound infections, and three minor wound breakdowns successfully treated. Hardware complications included three cervical plate/screw extrusions reoperated, one cervical plate fracture reoperated, one posterior cervical screw backout not reoperated, one case of broken pedicle screw not reoperated, one vertebral body failure not reoperated, and one posterior rod case reoperated for excessive rod length and protrusion. The overall complication rate attributable to placement of spinal instrumentation was 10/299 (3%) with a reoperation rate of 2%. The arthrodesis rate was 298/299 (99%). CONCLUSION: The complication rate for using spinal instrumentation can be less than previously reported. Lessons learned and discussed should reduce the rate even more. Spinal instrumentation is a safe and useful adjunct to fusion in treating degenerative, traumatic, infectious, and neoplastic diseases of the spine. PMID- 9400638 TI - Symptomatic pneumocephalus after transsphenoidal surgery. AB - BACKGROUND: Symptomatic pneumocephalus after transsphenoidal surgery, though reported, is a rare phenomenon. We report three cases of pneumocephalus in a series of 300 transsphenoidal operations for sellar/suprasellar mass lesions done over the past 12 years. METHODS AND RESULTS: Three cases of symptomatic pneumocephalus occurring after transsphenoidal surgery are presented to illustrate the causative factors, methods of prevention, and management. In case 1, an intraoperative cerebrospinal fluid (CSF) leak occurred and drainage of CSF through a lumbar subarachnoid drain resulted in pneumocephalus, in spite of repair of the sellar floor. In case 2, partial excision of tumor and subsequent reduction of intracranial pressure due to a ventriculoperitoneal (VP) shunt led to pneumocephalus. In case 3, radiotherapy-induced shrinkage of a partially excised tumor resulted in pneumocephalus. The sellar floor had not been repaired in cases 2 and 3 as there was no intraoperative CSF leak and only a partial excision had been done. Conservative management failed in the two patients in whom it was tried. Repair of the sella and sphenoid sinus had to be done in all three cases. CONCLUSIONS: Repair of the sellar floor should be done after a transphenoidal approach in all cases, even when no intraoperative leak has been identified and only a partial excision of tumor has been done. Once pneumocephalus has occurred, the sellar floor and sphenoid sinus should be repaired early before reducing the intracranial pressure (ICP) by tapping ventricular air and draining or diverting CSF. PMID- 9400639 TI - Prediction of consistency of meningiomas with preoperative magnetic resonance imaging. AB - BACKGROUND: The consistency of a meningioma is one of the important factors in determining the surgical outcome. If the surgeon is aware of the consistency of a meningioma preoperatively, the surgical plans will be influenced. A few papers have described the correlation between consistency of meningiomas and their magnetic resonance imaging (MRI) findings. However, prediction of consistency with MRI is still difficult. We have tried to predict the consistency of meningiomas with MRI findings more precisely. METHODS AND RESULTS: Fifty patients diagnosed as having intracranial meningiomas were studied with 1.5 Tesla MRI. We compared the MRI findings with tumor consistency. The intensities of the tumors were categorized into three grades (low, iso, and high) compared to that of the gray matter. T1-weighted images had no specifics, but T2-weighted images and proton density images were useful for the prediction of tumor consistency. Hyperintensity on protein density (PD) and T2-weighted images was a sign of a soft tumor. CONCLUSION: We presume that T2 and PD are useful for predicting consistency of meningiomas, and their water content is one of the main factors in their consistency. Histology may be one of the factors helpful in defining the consistency of a tumor. In this series, we found no relationship between histology and MRI findings, nor between histology and consistency. If the meningioma is believed to be hard, preoperative endovascular embolization is beneficial, which will induce necrosis of the meningioma and make it soft enough to be removed more easily and safety. PMID- 9400640 TI - Pathologic significance of meningeal enhancement ("flare sign") of meningiomas on MRI. AB - BACKGROUND: The purpose of this study was to clarify the pathologic features and clinical significance of the meningeal enhancement surrounding meningiomas ("flare sign") on contrast-enhanced T1-weighted magnetic resonance images (MRI). METHODS: The marginal dura mater of tumors was resected from nine cases of meningioma exhibiting a flare sign and used for histopathologic evaluation. RESULTS: Connective tissue proliferation was found in the dura mater in all cases, vascular proliferation was found in three, and tumor cell nests were observed in four cases. In one case, tumor cells were found 4.5 mm from the edge of the tumor. In another case, a meningothelial cell cluster was found. CONCLUSIONS: These results suggest that tumor cell nests are present frequently in dura mater that exhibits the flare sign, and that the dura mater near these lesions should be resected as widely as possible. PMID- 9400641 TI - Resection of a recurrent parasagittal meningioma with cortical vein anastomosis: technical note. AB - BACKGROUND: Simpson Grade I resection of parasagittal meningiomas is not always feasible because of the involvement of the sagittal sinus and cortical veins. Complete resection requires reconstruction of the sagittal sinus and cortical veins. This report describes a surgical technique to preserve patency of the cortical veins. CASE REPORT: A recurrent parasagittal meningioma completely occupied the superior sagittal sinus and encased several large cortical veins. The tumor in the sagittal sinus was totally resected and the roof of the sinus was sutured. To avoid thrombotic cortical vein occlusion, two cortical veins encased by the meningioma were anastomosed end-to-end, regardless of their flow directions. The postsurgical course was uneventful and patency of the anastomosed veins was confirmed by postoperative angiography. CONCLUSIONS: End-to-end anastomosis of cortical veins was a useful surgical technique for radical resection of a parasagittal meningioma. PMID- 9400642 TI - Classification and treatment of vertebral dissecting aneurysm. AB - BACKGROUND: For many years, dissecting aneurysms of the intracranial vertebral artery were believed to be quite rare. In recent years, because vascular disorders have been studied more thoroughly by three dimensional-computed tomography (3D-CT), angiographically and pathologically, these aneurysms are being reported with more frequency. METHODS: Among the 45 patients diagnosed to have aneurysms arising from the vertebral artery or its branches over a 20-year period, 16 had dissecting aneurysms. The authors present their therapeutic strategy for these patients. Surgery was performed in the 16 patients, the most common technique being clip-occlusion or trapping of the parent artery wherever feasible, in an attempt to optimize cerebral blood flow. The dissecting aneurysms of the vertebral artery were classified into two groups for the purpose of determining a therapeutic approach, namely unilateral and circumferential groups. In the unilateral group, the dissection seemed to involve only on one side of the vessel according to the conventional cerebral angiogram. These patients underwent surgical reconstruction of the vertebral artery by direct clipping. In the circumferential group, the dissection was all around the artery. Proximal clipping or trapping was performed in this group. RESULTS: In six out of eight patients with unilateral dissecting aneurysms, vascular reconstruction was possible by direct clipping. Of these six patients, the surgical outcome was considered excellent in four, fair in one, and one patient died of cardiac failure after 12 days as his preoperative morbid condition remained the same after surgery. Two other patients with unilateral dissecting aneurysms were treated with trapping technique and the surgical outcome was excellent in one patient and good in the other patient. Both patients resumed a normal social life. In five out of eight patients with circumferential dissecting aneurysms, trapping or proximal clipping was performed and the surgical outcome was excellent in two patients, good in one and fair in one patient. One patient with preoperative brain stem infarction died of aspiration pneumonitis after 8 months. Two patients who were noted to have an increase in the size of aneurysm during follow-up angiography underwent a craniotomy with clipping and wrapping of the aneurysm. There was a favorable surgical outcome in both patients. The remaining three patients had Grade IV subarachnoid hemorrhage (SAH) prior to surgery and at autopsy a disturbed vascular wall was detected. CONCLUSION: The authors' experience suggests that when surgically feasible, direct clipping is an effective alternative approach in the treatment of dissecting aneurysms of the vertebral artery in which blood flow in the parent artery is to be preserved. PMID- 9400643 TI - Spontaneous occlusion of a giant basilar tip aneurysm and a basilar artery due to the dissection of both structures: case report. AB - BACKGROUND: Spontaneous occlusion of a giant aneurysm with its parent artery is relatively rare. Complete occlusion of a giant aneurysm at the basilar bifurcation and a basilar artery due to the dissection of the basilar artery has never been reported. CASE DESCRIPTION: This 62-year-old man presented with left hemiparesis and right oculomotor palsy. Radiographic study showed a giant aneurysm at the basilar artery bifurcation with hemorrhage in its wall and an ischemic area in the right midbrain. Subsequent study revealed that thrombosis of the aneurysm rapidly progressed and that the parent basilar artery caused the dissection. Finally the giant aneurysm and the basilar artery were completely thrombosed. CONCLUSION: The dissection was considered to occur in the aneurysm wall first by the hemorrhage in it and progress proximally along the basilar artery. Intramural hemorrhage in the wall of a giant aneurysm can be a cause of dissection of its parent artery. This seems to be one of the mechanisms by which a giant aneurysm and its parent artery are spontaneously thrombosed. PMID- 9400644 TI - Brain cavernoma: a dynamic lesion. AB - BACKGROUND: Although the prevalence of brain cavernomas is high (0.50%), for unknown reasons, only a few of them display aggressive clinical behavior. METHODS: From a personal series of 65 operated and histopathologically verified cavernomas, we have conducted a long-term study, both retrospectively and prospectively, of the main features that cause some cavernomas to be dynamic lesions. RESULTS: Hemorrhage is the most common phenomenon. Extralesional bleeding due to the rupture of peripheral caverns is most often observed. These are never as immediately devastating as hemorrhages originating from a high-flow, high-pressure AVM. Extralesional hemorrhages tend toward spontaneous resorption, but the risk of recurrence exists and may lead to permanent disability or death (especially when the lesion is located in the brain stem). Intralesional bleeding caused by rupture of contiguous caverns is less frequently observed. This may lead to the formation of large cysts. Calcifications are mostly observed in patients presenting with chronic epilepsy. The bleeding risk of calcified cavernomas is low, but it can exist and should be taken into account in the surgical decision making. The growth of the cavernomatous matrix was obvious in three large cavernomas (two with calcification). No bleeding was found inside the lesions, suggesting a pure "intrinsic" growth. The role of pathologic angiogenic factors is highly probable in these cases. "De novo" appearing lesions were observed in five cases (four belonging to familial forms) on the magnetic resonance imaging survey of operated patients. Perilesional atrophy was observed in three cases (two operated) in patients with a long-lasting evolution. It suggests that the brain metabolism can be disturbed by slow, chronic effusion of blood around the cavernoma. CONCLUSIONS: The dynamism of cavernomas is determined by extrinsic factors, mainly hemorrhage (with its own consequences); and by intrinsic factors: the pseudotumoral growth of the cavernous matrix. Therefore, when they are symptomatic, cavernomas should be totally removed. PMID- 9400645 TI - Anastomosis of the superficial temporal artery to the middle cerebral artery with the interposed occipital artery graft in moyamoya disease: case report. AB - BACKGROUND: Although there have been various interposed bypass grafts used for cerebral revascularization, the occipital artery has never been used as a graft. Interposed occipital artery bypass graft in an adult case with moyamoya disease after failed indirect revascularization is presented. CASE DESCRIPTION: This 34 year-old woman with moyamoya disease, who had suffered from cerebral ischemic symptoms since the age of 6 years, was admitted to our hospital because of an intracerebral hemorrhage on the left side. She had undergone superficial temporal to-middle cerebral artery anastomosis, encephalo-galeo-synangiosis on the right side, and encephalo-duro-arterio-synangiosis on the left side at age 29 years. Four months after the intracerebral hemorrhage, she still had cerebral ischemic symptoms in the left hemisphere where cerebral revascularization was poor. Since neither the superficial temporal nor occipital artery could be used for direct anastomosis because of spontaneous transdural anastomoses of the superficial temporal artery and the short length of the occipital artery, anastomosis between the left superficial temporal artery and left posterior parietal artery was performed using a left occipital artery graft 6 months after the hemorrhage. Postoperative external carotid angiograms showed good patency of the graft. CONCLUSION: In cases in which direct anastomosis is infeasible for cerebral revascularization, the occipital artery could successfully be used as a bypass graft. PMID- 9400646 TI - Vasoreconstructive surgery for radiation-induced vasculopathy in childhood. AB - BACKGROUND: Cerebral vasculopathy associated with the appearance of netlike vessels can develop following irradiation therapy for parasellar brain tumors, especially in children. However, little is known regarding the clinical course of this disease or the appropriate therapy for it. CASE REPORTS: We experienced two surgically-treated patients with radiation-induced vasculopathy and reviewed the previously reported cases. Both of our patients were treated with encephalo-duro arterio-myo-synangiosis combined with superficial temporal artery to middle cerebral artery (STA-MCA) bypass. Their ischemic symptoms improved following the surgery, associated with a good angiographic neovascularization from the STA-MCA bypass, as well as dural and muscular arteries. CONCLUSION: Our findings and the review of the previous reports suggested that surgical therapy may be beneficial for the patients with radiation-induced vasculopathy with the appearance of netlike vessels. PMID- 9400647 TI - Transcondylar approach for dural arteriovenous fistulas of the cervicomedullary junction. AB - BACKGROUND: Spinal dural arteriovenous fistulas are abnormal arteriovenous connections on the surface of the dura. The site of the fistula is most commonly in the thoracic and lumbosacral regions and they are rarely located in the cervical region. CASES: The patients had two asymptomatic dural arteriovenous fistulas of the cervicomedullary junction fed by the left posterior meningeal artery and draining to the dilated coronal venous plexus and the radiculomedullary vein. RESULTS: The lesions were successfully treated by surgical interruption of the intrathecal vein with coagulation via a suboccipital transcondylar approach and a condylar fossa approach. Both patients left the hospital without significant deficits. CONCLUSIONS: We recommend that dural arteriovenous fistulas in the cervical region be surgically treated. PMID- 9400648 TI - Cranial titanium osteosynthesis systems. PMID- 9400649 TI - Pitfalls of facial nerve enhancement on MRI. PMID- 9400650 TI - The Visible Human Project. PMID- 9400651 TI - Reducing the costs of health care: the other side of the equation. PMID- 9400652 TI - Migration of nitrosamines from condoms to physiological secretions. PMID- 9400653 TI - Elevated dentine lead levels in adult teeth of First Nation people from an isolated region of northern Ontario, Canada. PMID- 9400654 TI - Trace element pollution of soils collected near a municipal solid waste incinerator: human health risk. PMID- 9400655 TI - Vapor phase and particulate bound polycyclic aromatic hydrocarbons in the smoke of mosquito coils. PMID- 9400656 TI - HCH and DDT residues in drinking water from the South of Spain, 1991-1994. PMID- 9400657 TI - Volatilization of arsenic in contaminated cattle dipping vat soil. PMID- 9400658 TI - Effect of antioxidants on UV-induced DNA breakage in human peripheral lymphocytes. PMID- 9400659 TI - Effect of methacrylonitrile on rat lung antioxidant enzymes. PMID- 9400660 TI - Biodisposition study of the organophosphorus pesticide, methyl-parathion. PMID- 9400661 TI - Determination of pentachlorophenol in environmental samples of the S. Euboic Gulf, Greece. PMID- 9400662 TI - Monitoring of the pesticide levels in natural waters of Greece. PMID- 9400663 TI - Phototransformation of 2-chloroaniline in aqueous solution. PMID- 9400664 TI - Biodegradation of endosulfan by a bacterial coculture. PMID- 9400665 TI - Adsorption characteristics of trihalomethanes onto activated carbon fiber from quarternary mixture solution. PMID- 9400666 TI - Improved blue rayon hanging technique that can measure a time-weighted average concentration of benzo(a)pyrene in sea water. PMID- 9400667 TI - Response of the stonefly Pteronarcys dorsata in enclosures from an urban North Carolina stream. PMID- 9400668 TI - Sensitivity of chironomus plumosus larvae to V5+, Mo6+, Mn2+, Ni2+, Cu2+, and Cu+ metal ions and their combinations. PMID- 9400669 TI - Comparative acute toxicity of some pesticides, metals, and surfactants to Gammarus italicus Goedm. and Echinogammarus tibaldii pink. and stock (Crustacea: Amphipoda). PMID- 9400670 TI - Bioconcentration of chlorpyrifos, chlorfenvinphos, and methidathion in Mytilus galloprovincialis. PMID- 9400671 TI - Species variation and some properties of renal glutathione S-transferase of fish from Arabian gulf. PMID- 9400672 TI - Comparative study on the acute toxicities of alpha, beta, gamma, and delta isomers of hexachlorocyclohexane to freshwater fishes. PMID- 9400673 TI - Effect of naphthalene on carbohydrate metabolism during vitellogenesis in marine edible crab, Scylla Serrata. PMID- 9400674 TI - Effect of selenium on mercury methylation in anaerobic lake sediments. PMID- 9400675 TI - Assay for toxic chemicals using bacteria. PMID- 9400676 TI - Gastro-oesophageal reflux disease and asthma. AB - Gastro-oesophageal reflux disease (GOR) and asthma are both common medical conditions that often co-exist. Studies using oesophageal manometry and 24 h ambulatory pH monitoring have shown that up to 80% of asthmatics have abnormal GOR. A number of mechanisms whereby GOR may trigger asthma have been proposed, and it is believed that acid reflux may stimulate vagal receptors in the lower oesophagus causing reflex bronchoconstriction. However, GOR may be worsened by asthma causing abnormal diaphragm mechanics and by its treatment. Formal evaluation of GOR should be considered a part of asthma assessment, particularly if asthmatic symptoms are precipitated by factors known to trigger GOR such as reclining, alcohol ingestion, and the use of theophylline. Twenty-four hour ambulatory intra-oesophageal pH monitoring remains the gold standard for the diagnosis of GOR. Medical therapy with anti-reflux medications, such as acid suppressive agents and prokinetic agents may improve both GOR and asthma control. In those who fail medical therapy, anti-reflux surgery may be warranted in some. PMID- 9400677 TI - Environment and the ageing lung. AB - A brief review of changes in the morphology, physiology and defence mechanism in the ageing lung is presented. During the ageing process, the lung is also affected by environmental insults, which can be exogenous or endogenous. Exogenous factors include infection, climate, air pollution and mechanical injuries, whereas endogenous environments are certain system diseases (e.g. diabetes mellitus and thromboembolism) as well as infectious diseases (e.g. tuberculosis). PMID- 9400678 TI - Non-invasive ventilation in the management of respiratory failure. AB - Non-invasive positive pressure ventilation (NIPPV) has been used increasingly to treat various forms of respiratory failure, with benefits in terms of gas exchange improvement, avoidance of endotracheal intubation and a decreased mortality. This review will focus on the recent developments and recommendations in the use of NIPPV in the treatment of acute and chronic respiratory failure, the methodology in the application of NIPPV, and briefly on its proposed mechanism of action. PMID- 9400679 TI - Surgical results of 29 patients with benign tracheobronchial lesions. AB - This study was carried out in order to evaluate the surgical results of benign tracheobronchial diseases. Between July 1988 and March 1996, tracheobronchial surgery was performed on 29 patients with a variety of benign diseases. The primary diseases were post intubation or post tracheostomy tracheal stenosis (n = 12), tuberculous stenosis (n = 7), congenital tracheal stenosis with or without vascular ring (n = 4), tracheobronchial tumour (n = 2), oesophageal tumour (n = 1), and miscellaneous conditions (n = 3). Thirty-one operative procedures included sleeve lobectomy (n = 7), sleeve resection of trachea (n = 17) and bronchus (n = 2), and plastic surgery of trachea (n = 4) and bronchus (n = 1). There was one operative death, which put the mortality rate at 3.4%. There were five postoperative complications in this series (17.2%), including anastomotic disruption of trachea (n = 1), bilateral vocal cord palsy (n = 1), prolonged endotracheal intubation (n = 1) and overgrowth of granulation (n = 2). The complication of anastomotic disruption of trachea was treated by insertion of a tracheal T-tube, and the granulation was treated by bronchoscopic excision. We suggest that tracheobronchoplasty is a safe procedure in carefully selected patients with benign diseases. PMID- 9400680 TI - Hepatocyte growth factor promotes growth and lumen formation of fetal lung epithelial cells in primary culture. AB - Mesenchyme-epithelium interactions are generally considered critical for fetal lung development. Hepatocyte growth factor (HGF), a mesenchyme-derived mitogen active on a variety of epithelial cells, appears to be involved in the morphogenesis of fetal liver and kidney. During lung development, HGF and its receptor, c-Met, are expressed in close proximity in mesenchymal cells and epithelial cells, respectively. To examine the role of HGF in fetal lung development, we investigated the effects of HGF on lung epithelial cells derived from a 15-day-old mouse fetus. First, HGF induces a 45% increase in [3H]thymidine incorporation and a 65% increase in cell number by crystal violet analysis at 10 ng/mL concentration, and the increase is dose dependent. Second, HGF facilitates the formation of an organotypic arrangement of the fetal epithelial cells on a basement membrane extract (Matrigel) that resembles alveolar structures in vivo, and the maximum increase is about twice the control level at 10 ng/mL. These results suggest that HGF may be implicated in fetal lung development through the regulation of mesenchyme-epithelium interactions. PMID- 9400682 TI - A case of malignant pleural mesothelioma following exposure to atomic radiation in Nagasaki. AB - We report the case of a 75-year-old Japanese man who developed malignant mesothelioma in the left hemithorax 50 years after the dropping of the atomic bomb on Nagasaki in 1945. This may be the first reported case of malignant mesothelioma following exposure to atomic radiation. Asbestos is the leading cause of malignant mesothelioma, but radiation therapy is the primary non asbestos-related cause. In the case of radiation therapy, the interval between exposure and the occurrence of malignant mesothelioma tends to be many years. This patient was at a high risk of malignant mesothelioma as he had been exposed to radiation from the atomic bomb and may also have had a history of asbestos exposure at the munitions factory where he was employed as a shipbuilder for 2 years. It has been suggested that combined exposure to atomic radiation and asbestos is associated with an increased incidence of malignant mesothelioma. If thickening of the pleura or pleural effusion is found in atomic bomb survivors, malignant mesothelioma should be considered as one of the options in the differential diagnosis, even although the atomic bomb attacks occurred several decades ago. PMID- 9400681 TI - Tachykinins contribute to the acute airways response to allergen in sheep actively sensitized to Ascaris suum. AB - Tachykinins, found in sensory nerves, have effects in the airways which suggest that they may contribute to the pathogenesis of asthma. We aimed to find evidence for tachykinin involvement in the immediate airway response to allergen in a sheep model of experimental asthma. Twenty-four sheep were actively sensitized to Ascaris suum, then challenged with nebulized Ascaris extract in a dose-response fashion. Change in lung resistance (RL) in response to challenge was measured. Responder sheep (those with an increase in RL of > or = 100% over baseline) that had reproducible responses over three challenges were identified (n = 4 sheep) and a PC100 (number of breaths of extract required to induce a 100% increase in RL) was determined. The effect of the neutral endopeptidase inhibitor phosphoramidon, the NK-1 receptor-specific antagonist CP 96, 345 and capsaicin desensitization on the RL response to Ascaris challenge was then assessed. Administration of phosphoramidon before Ascaris decreased the PC100 to 31 +/- 7% of the PC100 seen with Ascaris alone (P < 0.05), whereas CP 96,345 and capsaicin desensitization increased the PC100 to 285 +/- 41% and 555 +/- 93% respectively (P < 0.05 for both). These findings suggest that endogenous tachykinins are released in response to allergen challenge and that they contribute to the immediate increase in RL. PMID- 9400683 TI - Tuberculosis and HIV infection: global perspectives. AB - This paper reviews the epidemiological and clinical aspects of the interaction between Mycobacterium tuberculosis and HIV infection. The incidence of HIV associated tuberculosis is increasing worldwide and is expected to increase further, especially in Africa and parts of Asia. HIV infection appears to increase the likelihood that tuberculous infection will occur after tubercle bacilli are inhaled into the lungs. Moreover, there is persuasive evidence that in the presence of HIV infection, new-onset tuberculous infection will progress rapidly to clinically significant disease and the probability that latent tuberculous infection will reactivate is enormously increased. The accelerating and amplifying influence of HIV infection is also contributing to the increasing incidence of disease caused by multidrug-resistant strains of M. tuberculosis. Neither clinical nor radiographic features reliably distinguish the majority of patients with HIV-associated tuberculosis from those who are non-HIV-infected. Some HIV-infected patients, however, have atypical manifestations and are difficult to diagnose. Chemotherapy for 6 months with conventional antituberculosis drugs cures most patients, but many died during or after treatment of other AIDS-related complications. HIV is contributing heavily to the worldwide increase in tuberculosis. There is also mounting evidence that tuberculosis accelerates the course of co-existing HIV disease. PMID- 9400684 TI - Current issues in the management of chronic obstructive pulmonary diseases. AB - Chronic obstructive pulmonary disease (COPD) is a leading cause of morbidity and mortality, especially among smokers. Many guidelines that have recently been issued emphasize that COPD is not inaccessible to therapeutic measures: although few interventions are capable of affecting its natural history (i.e. smoking cessation and, in patients with severe resting hypoxaemia, oxygen therapy), several others have a demonstrated effect on symptoms and, thereby, quality of life. The effects of inhaled corticosteroids, and alpha 1-antitrypsin replacement therapy in emphysema due to alpha 1-antitrypsin deficiency are currently being studied. When there is a marked increase in mucus production, chest physiotherapy using controlled expiration and directed cough may be useful. Inhaled bronchodilators are frequently effective on dyspnoea, anticholinergic agents being more suitable for continuous symptoms. Rehabilitation, which includes education and psychosocial care, chest physiotherapy, nutritional care and exercise training, also improves quality of life. When there is persistent severe alveolar hypoventilation despite oxygen therapy, long-term mechanical ventilation may be considered. Surgical options in the treatment of emphysema include resection of giant bullae and lung volume reduction surgery. Lung transplantation should be proposed only in patients with end-stage disease, the difficulty here being to define what 'end-stage' means. Finally, all preventive and some therapeutic interventions are likely to be more effective early in the course of the disease. Thus, efforts should be made to detect airways obstruction early in subjects at risk, such as smokers. PMID- 9400685 TI - Applications of colour Doppler ultrasound in the diagnosis of chest diseases. AB - Colour Doppler ultrasound (US) or colour Doppler imaging is a new imaging modality that can simultaneously display blood flow information and Doppler spectral analysis. This new technique provides an opportunity to assess pulmonary blood flow and perfusion non-invasively. Colour Doppler US has many potential applications in diseases of the chest. Colour Doppler imaging is useful in assisting the diagnosis of pulmonary arteriovenous malformation and pulmonary sequestration. The 'fluid colour sign' can be used to detect minimal effusion amenable to thoracentesis. Colour Doppler US can be used to assess the angiogenesis of a lung tumour and may be helpful in differentiating a malignant tumour from a benign one. Colour Doppler US can be used to guide a transthoracic needle biopsy and improve the safety of this invasive procedure. Colour Doppler US can demonstrate the vascular patterns and assess the regional haemodynamic changes of a pulmonary consolidation. The information of spectral wave analysis is helpful for understanding the haemodynamic changes of a pulmonary consolidation. Colour Doppler US is useful in assessing perfusion and reperfusion status of a pulmonary infarction. The recent advent of amplitude US angiography further improved the sensitivity of colour Doppler US in detecting blood flow signal without angle restriction. The potential application of these new techniques in chest diseases may need further exploration. PMID- 9400686 TI - Role of endogenous nitric oxide in the pathogenesis of bronchial asthma. PMID- 9400687 TI - ATS standards for the optimal management of chronic obstructive pulmonary disease. AB - Tobacco smoking is the main cause of chronic obstructive pulmonary disease (COPD), and the provision of encouragement and support in smoking cessation is the best way to help patients with COPD. The three major goals of COPD management are to lessen airflow limitation, to prevent and treat secondary medical complications and to decrease respiratory symptoms and improve quality of life. Outpatient pharmacotherapy should be organized in a stepwise manner according the severity of disease, the aims being to induce bronchodilation, reduce inflammation and facilitate expectoration, although the role of anti-inflammatory and mucolytic treatment in COPD has not been clearly established. Patients who are not well controlled on optimal pharmacotherapy are candidates for enrollment in a pulmonary rehabilitation programme. Correction or prevention of hypoxaemia is a priority, and long-term oxygen therapy may prolong survival in hypoxaemic patients. With only limited data on criteria for hospital admission and the objectives of hospitalization, the published ATS standards on the management of COPD include expert consensus statements on these aspects of hospital care. Special considerations, such as surgery in the COPD patient, sleep disorders, nutrition, air travel, and ethical issues, are discussed. PMID- 9400688 TI - Efficacy and safety of inhalation therapy in chronic obstructive pulmonary disease and asthma. AB - The inhaled route has a number of well-recognized advantages over other routes for administration of drugs to the lungs. As the drug is delivered directly to its required site of action, only a small quantity is required for an adequate therapeutic response. Consequently, there is a low incidence of systemic side effects compared with oral or intravenous administration. In addition, the onset of action of inhaled drugs is generally faster than that achieved by oral administration. Factors that can affect the quantity of inhaled particles reaching the lungs and their topographical distribution are manner of inhalation, aerosol characteristics and subject characteristics. The main types of inhaler device are metered-dose inhalers (MDIs), dry powder inhalers (DPIs) and nebulizers (jet and ultrasonic). There are advantages and disadvantages associated with all types of currently available inhaler device and there is still a clear need to develop more efficient devices that are easy to use. Respimat (Boehringer Ingelheim) is a novel soft mist inhaler that operates via a mechanism utilizing mechanical power rather than gas propellants, and represents an exciting development in this field. PMID- 9400689 TI - Choice of bronchodilator therapy in chronic obstructive pulmonary disease. AB - The majority of patients with chronic obstructive pulmonary disease may derive benefit from bronchodilator therapy, particularly those patients with severe disease. Currently, three classes of bronchodilator are available: anticholinergics, beta 2-adrenergic agonists and methyl-xanthines. Several factors should be considered when choosing a bronchodilator. Inhaled therapy is generally preferred over oral treatment as the inhaled route reduces systemic exposure to the drug, whereas oral therapy has the advantages of convenience and ease of use. The anticholinergics (e.g. ipratropium bromide) and beta 2-agonists (e.g. salbutamol) are used via the inhaled route and have approximately equivalent acute bronchodilator effects; they differ pharmacokinetically in onset of effect and duration of action and the anticholinergic may be superior in its long-term effects on lung function. PMID- 9400690 TI - Combined anticholinergic therapy in the management of acute asthma. AB - In a hospital setting, first-line drugs for treatment of acute severe asthma are usually nebulized short-acting beta 2-agonists and systemic corticosteroids. Combining a nebulized beta 2-agonist with the anticholinergic agent ipratropium bromide may produce better bronchodilation than either drug alone, particularly in patients with more severe episodes. A recent study from New Zealand compared nebulized beta 2-agonist alone or in combination in patients presenting with acute severe asthma to the emergency department. Combined treatment produced a significantly greater change in forced expiratory volume in 1 s (FEV1) than salbutamol alone. Factors predicting a poor bronchodilator response (irrespective of study drug received) were frequent use of inhaled beta 2-agonist before presenting at the emergency department, increased severity and duration of attack and older age. Patients presenting with more severe asthma had taken more inhaled bronchodilator before presentation and were therefore likely to be higher on the dose-response curve and thus less likely to derive benefit from additional bronchodilator therapy. The patients most likely to benefit from the addition of nebulized ipratropium bromide were those who had taken less inhaled beta 2 agonist in the previous 6 h. A combined analysis of three large studies on anticholinergic therapy, including the New Zealand study, has shown a 17% reduction in risk of subsequent admission (R = 0.83, 95% CI 0.63-1.1). Thus, nebulized ipratropium bromide imparts a small improvement in lung function when compared with salbutamol alone; however, further studies are needed to determine if multiple doses of combined anticholinergic/beta 2-agonist treatment reduce need for admission. PMID- 9400691 TI - Combined bronchodilator therapy in the management of chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is a leading cause of death and disability. There is now a better understanding of the pathophysiology of COPD and of the effectiveness of various treatment strategies in controlling symptoms and progression of disease. Although cessation of smoking is of primary importance, the growing realization in recent years that airflow limitation in COPD can be significantly relieved with the use of bronchodilators has changed the clinical approach to treating this disease. As a result, inhaled bronchodilators have become the therapy of choice in new pharmacological treatment algorithms for COPD. Additionally, the improvement in airflow limitation seen with bronchodilators, if maintained by patient compliance, can result in an improved level of lung function. Inhaled beta 2-adrenergic agonists are effective bronchodilators with a relatively rapid onset of action. They may also have additional value in that they can increase mucociliary clearance in the airways. Inhaled anticholinergic bronchodilators, such as ipratropium bromide, have been shown to be more effective bronchodilators than beta 2-agonists in COPD; they are associated with a low incidence of side-effects and may decrease the number of pulmonary exacerbations. The use of the combination of these two classes of inhaled bronchodilators provides superior bronchodilation than treatment with either of the individual components without added side-effects or loss of the positive effects of ipratropium bromide including reduced exacerbation frequency and lack of tachyphylaxis. The use of combination therapy also improves cost-effectiveness and patient compliance. Combination therapy should be considered as an important component of a treatment algorithm of COPD. PMID- 9400692 TI - Mineral properties and their contributions to particle toxicity. AB - It has been recognized since at least as early as the mid-1500s that inhaled minerals (i.e., inorganic particles) can pose a risk. Extensive research has focused on the biological mechanisms responsible for asbestos- and silica-induced diseases, but much less attention has been paid to the mineralogical properties and geochemical mechanisms that might influence a mineral's biological activity. Several important mineralogical characteristics control a mineral's reactivity in geochemical reactions and are likely to determine its biological reactivity. In addition to the traditionally considered variables of particle size and shape, mineralogical characteristics such as dissolution behavior, ion exchange, sorptive properties, and the nature of the mineral surface (e.g., surface reactivity) play important roles in determining the toxicity and carcinogenicity of a particle. Ultimately, a mineral's species (which provides direct information on a mineral's structure and composition) is probably one of the most significant yet most neglected factors that must be considered in studies of toxicity and carcinogenicity. PMID- 9400694 TI - Chemical characterization and reactivity of iron chelator-treated amphibole asbestos. AB - Iron in amphibole asbestos is implicated in the pathogenicity of inhaled fibers. Evidence includes the observation that iron chelators can suppress fiber-induced tissue damage. This is believed to occur via the diminished production of fiber associated reactive oxygen species. The purpose of this study was to explore possible mechanisms for the reduction of fiber toxicity by iron chelator treatments. We studied changes in the amount and the oxidation states of bulk and surface iron in crocidolite and amosite asbestos that were treated with iron chelating desferrioxamine, ferrozine, sodium ascorbate, and phosphate buffer solutions. The results have been compared with the ability of the fibers to produce free radicals and decompose hydrogen peroxide in a cell-free system in vitro. We found that chelators can affect the amount of iron at the surface of the asbestos fibers and its valence, and that they can modify the chemical reactivity of these surfaces. However, we found no obvious or direct correlations between fiber reactivity and the amount of iron removed, the amount of iron at the fiber surface, or the oxidation state of surface iron. Our results suggest that surface Fe3+ ions may play a role in fiber-related carboxylate radical formation, and that desferrioxamine and phosphate groups detected at treated fiber surfaces may play a role in diminishing and enhancing, respectively, fiber redox activity. It is proposed that iron mobility in the silicate structure may play a larger role in the chemical reactivity of asbestos than previously assumed. PMID- 9400693 TI - Surface reactivity in the pathogenic response to particulates. AB - The peculiar characteristics of dust toxicity are discussed in relation to the processes taking place at the particle-biological medium interface. Because of surface reactivity, toxicity of solids is not merely predictable from chemical composition and molecular structure, as with water soluble compounds. With particles having the same bulk composition, micromorphology (the thermal and mechanical history of dust and adsorption from the environment) determines the kind and abundance of active surface sites, thus modulating reactivity toward cells and tissues. The quantitative evaluation of doses is discussed in comparisons of dose-response relationships obtained with different materials. Responses related to the surface of the particle are better compared on a per unit surface than per-unit weight basis. The role of micromorphology, hydrophilicity, and reactive surface cations in determining the pathogenicity of inhaled particles is described with reference to silica and asbestos toxicity. Heating crystalline silica decreases hydrophilicity, with consequent modifications in membranolytic potential, retention, and transport. Transition metal ions exposed at the surface generate free radicals in aqueous suspensions. Continuous redox cycling of iron, with consequent activation-reactivation of the surface sites releasing free radicals, could account for the long-term pathogenicity caused by the inhalation of iron-containing fibers. In various pathogenicities caused by mixed dusts, the contact between components modifies toxicity. Hard metal lung disease is caused by exposure to mixtures of metals and carbides, typically cobalt (Co) and tungsten carbide (WC), but not to single components. Toxicity stems from reactive oxygen species generation in a mechanism involving both Co metal and WC in mutual contact. A relationship between the extent of water adsorption and biopersistence is proposed for vitreous fibers. Modifications of the surface taking place in vivo are described for ferruginous bodies and for the progressive comminution of chrysotile asbestos fibers. PMID- 9400695 TI - Study of the stability of a paramagnetic label linked to mesoporous silica surface in contact with rat mesothelial cells in culture. AB - Stable radicals detectable by electron paramagnetic resonance (EPR) may be use in the investigation of early events in cell-particle toxicity. Piperidine-N-oxyl derivatives (nitroxides), covalently linked to the surface of a high surface area silica (used as model solid for the technique), served as probes in the investigation of the effects of incubation of silica particles with mesothelial cells. A mesoporous silica (MCM-41), prepared by precipitation from a micellar solution, was the most appropriate silica-based particle for this purpose, as its channels allow direct contact with small molecules but not with macromolecules. The cytotoxicity of this amorphous silica is very low, allowing relatively high particle loading in the cell cultures. Both the high surface area of the sample and the large amount of inorganic material extracted from the cell culture provide enough material to run reasonably intense EPR spectra. Computer-aided analysis of the EPR spectra of silica-bound nitroxides provided information on the sensitivity of the labeled silica monitoring different environments, e.g., to follow the path of particles in a mammalian cell culture. Upon contact of the particles with mesothelial cells, the mean distance among the labels at the silica surface decreased as a consequence of the release of oxidizing and/or radical moieties from the cells. PMID- 9400696 TI - Demonstration of nitric oxide on asbestos and silicon carbide fibers with a new ultraviolet spectrophotometric assay. AB - Nitric oxide (NO) has a number of important functions in biological systems and may play a role in the toxicity of mineral fibers. We investigated whether NO might be present on the surface of mineral fibers and if crocidolite could adsorb NO from NO gas or cigarette smoke. NO was determined with a new gas chromatography-ultraviolet spectrophotometric technique after thermal desorption from the fiber surface and injection in a gas flow cell. NO was found in different amounts on chrysotile B, crocidolite, amosite, and silicon carbide whiskers. There was a strong correlation between the amount of NO and the specific surface area of these fibers (r = 0.98). NO could not be demonstrated on rockwool fibers [man-made vitreous fiber(s) (MMVF)21 and MMVF22] or silicon nitride whiskers. NO on crocidolite, amosite, and silicon carbide whiskers was readily desorbed from the fibers at increased temperature, while NO on chrysotile B seemed to be more firmly adsorbed to the fiber and required a longer period of time to be desorbed. The amount of NO bound to crocidolite increased from 34 micrograms/g fiber to 85 and 474 micrograms/g after exposing the fibers to cigarette smoke and NO gas, respectively. These findings indicate that a) NO adsorbs to fiber surfaces, b) some fibers adsorb more NO than others, c) some fibers adsorb NO more strongly than others, and d) the amounts of NO on fibers may be increased after exposure of the fiber to cigarette smoke or other sources of NO. The biological significance of NO on mineral fibers remains to be investigated. PMID- 9400697 TI - Can microwave radiation at high temperatures reduce the toxicity of fibrous crocidolite asbestos? AB - Exposure of animals and humans to crocidolite asbestos fibers produces fibrosis and two types of cancers: bronchogenic carcinoma and mesothelioma. It is therefore desirable to reduce toxicity of these fibers without affecting their other characteristics. In this study, commercial crocidolite asbestos fibers were radiated with microwave radiation at different temperatures. Radiated fibers and nonradiated original fibers were then studied by Mossbauer spectroscopy to quantify the amount of ferric and ferrous ions present at structurally different sites in each crocidolite sample. They were also studied for their ability to initiate the peroxidation of linoleic acid to assess the effect of radiation on this process. Results showed that microwave radiation reduced the total Fe2+/Fe3+ ratio. This reduction produced a concomitant decrease in the ability of the radiated samples to peroxidize linoleic acid. PMID- 9400698 TI - Significance of the biodurability of man-made vitreous fibers to risk assessment. AB - It is generally agreed that the biodurability of man-made vitreous fibers is a major factor for the characterization of potential health effects. As there is currently no standardization of experimental protocols to determine biodurabilty, the results of the clearance assays have not been used up to now for regulatory purposes. Methods used to analyze biodurability in animal models are short-term inhalational exposure and intratracheal instillation of rat respirable fibers. Both test methods have strengths and limitations for regulatory purposes. We outline recommended procedures for standardized biodurability assays that can be used to compare different fiber types. In animal experiments, biodurability is difficult to separate from biopersistence, as mucociliary and macrophage-mediated clearance occur simultaneously with dissolution and disintegration. For intratracheal instillation, a sized rat respirable sample must be used. Precautions should be taken to prevent aggregation of fibers in the lungs. Although from a scientific point of view questions remain about quantifying the influence of fiber length, diameter, dose, and exposure route, consistent data on the biodurability of vitreous glass fibers are available which may be used for regulatory purposes. PMID- 9400699 TI - Investigations on health-related properties of two sepiolite samples. AB - Published i.p. injection studies have shown different biological behavior of different sepiolite samples. There was no evidence for carcinogenic potential of sepiolite from Vicalvaro, Spain, whereas a high tumor incidence was reported for sepiolite from Finland. The low biological activity of the sepiolite from Vicalvaro, compared to the Finnish sample, could be caused by low in vivo persistence or by the short length of the fibers, or both. In this study a further sepiolite sample, obtained as a commercial sample originating from China, was investigated. This sample contained a higher fraction of fibers longer than 5 microns, comparable to the Finnish sepiolite sample. The fraction of fibers with a length > 5 microns was 0.12 and 2.2% for the Vicalvaro and Chinese sepiolite, respectively. For the fiber fraction longer than 8 microns, the corresponding values were 0.0045 and 0.82%. The in vivo persistence of the sepiolite samples from China and Vicalvaro was analyzed after intratracheal instillation of 2 mg in female Wistar rats. Fiber retention in the lungs was analyzed by transmission electron microscopy at different sacrifice dates up to 12 months after application. For the Vicalvaro sepiolite, a splitting of fiber bundles was found during retention time in the lung. Therefore, no half-time of the fiber clearance could be calculated from the number of fibers. The decrease of the calculated retained fiber mass was faster for the Vicalvaro sepiolite (T1/2 = 89 days) compared to the Chinese sepiolite (T1/2 = 129 days). For 2 or 3 rats per group, at sacrifice date 12 months after i.p. injection, the lung was investigated by histopathology. The main difference between both treatment groups was a more pronounced fibrotic response in the Chinese sepiolite-treated rats compared to those treated with Vicalvaro sepiolite. It is concluded that both the higher fraction of long sepiolite fibers and the slower elimination rate of the fiber mass in the Chinese sample were important factors for the different biological reaction in comparison with Vicalvaro sepiolite. PMID- 9400700 TI - Bioavailable transition metals in particulate matter mediate cardiopulmonary injury in healthy and compromised animal models. AB - Many epidemiologic reports associate ambient levels of particulate matter (PM) with human mortality and morbidity, particularly in people with preexisting cardiopulmonary disease (e.g., chronic obstructive pulmonary disease, infection, asthma). Because much ambient PM is derived from combustion sources, we tested the hypothesis that the health effects of PM arise from anthropogenic PM that contains bioavailable transition metals. The PM samples studied derived from three emission sources (two oil and one coal fly ash) and four ambient airsheds (St. Louis, MO; Washington; Dusseldorf, Germany; and Ottawa, Canada). PM was administered to rats by intratracheal instillation in equimass or equimetal doses to address directly the influence of PM mass versus metal content on acute lung injury and inflammation. Our results indicated that the lung dose of bioavailable transition metal, not instilled PM mass, was the primary determinant of the acute inflammatory response for both the combustion source and ambient PM samples. Residual oil fly ash, a combustion PM rich in bioavailable metal, was evaluated in a rat model of cardiopulmonary disease (pulmonary vasculitis/hypertension) to ascertain whether the disease state augmented sensitivity to that PM. Significant mortality and enhanced airway responsiveness were observed. Analysis of the lavaged lung fluids suggested that the milieu of the inflamed lung amplified metal-mediated oxidant chemistry to jeopardize the compromised cardiopulmonary system. We propose that soluble metals from PM mediate the array of PM-associated injuries to the cardiopulmonary system of the healthy and at-risk compromised host. PMID- 9400701 TI - Oncogenes and tumor-suppressor genes in mesothelioma--a synopsis. AB - Invariably mesothelioma is diagnosed late in the development of the disease when treatment is no longer effective. Therefore, a key to reducing the mortality rate of this neoplasm is knowledge of the general sequence of genetic events between initiation of mesothelial cells and the emergence of the metastatic tumor cells. Unfortunately, relatively little is known about the early changes in the genesis of this disease. Of the known changes, the most frequent are in the tumor suppressor genes p16INK4a and NF2 and possibly the SV40 virus large T-antigen oncogene. The molecular nature of the changes in these genes as well as other alterations are addressed in this overview. PMID- 9400702 TI - Susceptibility of p53-deficient mice to induction of mesothelioma by crocidolite asbestos fibers. AB - Exposure of mesothelial cells to asbestos fibers in vitro has been shown to induce DNA damage mediated by oxidants. An early cellular response to DNA damage is increased expression of the p53 protein. This protein induces transcription of genes that activate cell cycle checkpoints or induce apoptosis. A murine mesothelial cell line that spontaneously acquired a point mutation in the p53 gene shows increased sensitivity to DNA damage induced by crocidolite asbestos fibers. It is hypothesized that p53-deficient mice will show increased sensitivity to the genotoxic effects of asbestos and accelerated development of malignant mesotheliomas. PMID- 9400704 TI - Malignant transformation of immortalized human bronchial epithelial cells by asbestos fibers. AB - Although asbestos is a well-established lung carcinogen, there currently is no suitable human cell model in which to examine the underlying cellular and molecular changes associated with fiber-mediated bronchial carcinogenesis. Using a recently established transformation model based on a human papillomavirus immortalized human bronchial epithelial cell line, we successfully transformed these BEP2D cells after a single, 7-day treatment with a 20-microgram/ml (4 micrograms per cm2 area) dose of Union Internationale Contre le Cancer (UICC) Rhodesian chrysotile fibers. Asbestos treatment resulted in a surviving fraction of 0.18 compared to control cells. Transformed cells developed through a series of sequential steps, including altered growth kinetics, resistance to serum induced terminal differentiation, and anchorage-independent growth, before becoming tumorigenic to form progressively growing tumors in nude mice. Seven tumorigenic cell lines were isolated and determined to be of human epithelial origin based on immunofluorescent staining of keratin and isozyme analysis. Analysis of tumor DNA revealed no mutations at either codon 12 or 13 in any the ras oncogenes. An independent role for K-ras mutation in fiber carcinogenesis, therefore, cannot be confirmed. This model provides a unique opportunity to study the cellular and molecular changes at the various stages in fiber-mediated neoplastic transformation of human bronchial epithelial cells. PMID- 9400703 TI - Mechanisms of fiber-induced genotoxicity. AB - The mechanisms of particle-induced genotoxicity have been investigated mainly with asbestos fibers. The results are summarized and discussed in this paper. DNA damage can be produced by oxidoreduction processes generated by fibers. The extent of damage yield depends on experimental conditions: if iron is present, either on fibers or in the medium, damage is increased. However, iron reactivity does not explain all the results obtained in cell-free systems, as breakage of plasmid DNA was not directly associated with the amount of iron released by the fibers. The proximity of DNA to the site of generation of reactive oxygen species (ROS) is important because these species have an extremely short half-life. Damage to cellular DNA can be produced by oxidoreduction processes that originate from cells during phagocytosis. Secondary molecules that are more stable than ROS are probably involved in DNA damage. Oxidoreduction reactions originating from cells can induce mutations. Genotoxicity is also demonstrated by chromosomal damage associated with impaired mitosis, as evidenced by chromosome missegregation, spindle changes, alteration of cell cycle progression, formation of aneuploid and polyploid cells, and nuclear disruption. In some of these processes, the particle state and fiber dimensions are considered important parameters in the generation of genotoxic effects. PMID- 9400705 TI - Neutrophils amplify the formation of DNA adducts by benzo[a]pyrene in lung target cells. AB - Inflammatory cells and their reactive oxygen metabolites can cause mutagenic effects in lung cells. The purpose of this study was to investigate the ability of activated neutrophils to modulate DNA binding of benzo[a]pyrene (B[a]P), a known carcinogen, in lung target cells. Equivalent numbers of rat lung epithelial cells (RLE-6TN cell line) and freshly isolated human blood neutrophils (PMN) were coincubated in vitro for 2 hr after addition of benzo[a]pyrene (0.5 microM) or two of its trans-diol metabolites, with or without stimulation with phorbol myristate acetate (PMA). DNA adducts of B[a]P-metabolites were determined in target cells using 32P-postlabeling; oxidative DNA damage (7-hydro-8-oxo-2' deoxyguanosine [8-oxodG]) was evaluated by high performance liquid chromatography with electrochemical detection. Increased DNA adducts were observed in lung cells coincubated with polymorphonuclear leukocytes (PMN). Activation of PMN with PMA, or addition of more activated PMN in relation to the number of lung cells, further increased the number of adducts, the latter in a dose-response manner. Incubation with B[a]P-4,5-diol did not result in any adduct formation, while B[a]P-7,8-diol led to a significant number of adducts. Moreover, PMA-activated PMN strongly enhanced adduct formation by B[a]P-7,8-diol, but not 8-oxodG, in lung cells. The addition of antioxidants to the coincubations significantly reduced the number of adducts. Results suggest that an inflammatory response in the lung may increase the biologically effective dose of polycyclic aromatic hydrocarbons (PAHs), and may be relevant to data interpretation and risk assessment of PAH-containing particulates. PMID- 9400706 TI - A new mechanism for DNA alterations induced by alpha particles such as those emitted by radon and radon progeny. AB - The mechanism(s) by which alpha (alpha) particles like those emitted from inhaled radon and radon progeny cause their carcinogenic effects in the lung remains unclear. Although direct nuclear traversals by alpha-particles may be involved in mediating these outcomes, increasing evidence indicates that a particles can cause alterations in DNA in the absence of direct hits to cell nuclei. Using the occurrence of excessive sister chromatid exchanges (SCE) as an index of DNA damage in human lung fibroblasts, we investigated the hypothesis that alpha particles may induce DNA damage through the generation of extracellular factors. We have found that a relatively low dose of alpha-particles can result in the generation of extracellular factors, which, upon transfer to unexposed normal human cells, can cause excessive SCE to an extent equivalent to that observed when the cells are directly irradiated with the same irradiation dose. A short lived, SCE-inducing factor(s) is generated in alpha-irradiated culture medium containing serum in the absence of cells. A more persistent SCE-inducing factor(s), which can survive freeze-thaw and is heat labile is produced by fibroblasts after exposure to the alpha-particles. These results indicate that the initiating target for alpha-particle-induced genetic changes can be larger than a cell's nucleus or even a whole cell. How transmissible factors like those observed here in vitro may extend to the in vivo condition in the context of a particle-induced carcinogenesis in the respiratory tract remains to be determined. PMID- 9400707 TI - Fiber-specific molecular features of tumors induced in rat peritoneum. AB - Molecular markers such as mutational spectra or mRNA expression patterns may give some indication of the mechanisms of carcinogenesis induced by fibers and other carcinogens. In our study, tumors were induced by application of crocidolite asbestos or benzo[a]pyrene (B[a]P) to rat peritoneum. DNA and RNA of these tumors were subjected to analysis of point mutations and to investigation of mRNA expression patterns. With both assays we found typical features depending on the type of carcinogen applied. The analysis of point mutations in the tumor suppressor gene p53 revealed mutations in the B[a]P-induced tumors. However, in the tumors induced by crocidolite asbestos that were of the same tumor type as those induced by B[a]P, mutations in p53 were not detectable. Every mutation detected on the DNA level causes an amino acid substitution within one of the functional domains of the tumor suppressor protein. Therefore, these mutations seem to be of biological relevance for tumor progression and indicate a difference in the carcinogenesis regarding the type of the carcinogenic substance. An additional specificity of crocidolite-induced tumors was detectable by analyzing the mRNA expression of the tumor suppressor gene WT1, which is known to be expressed in human mesothelial and mesothelioma cells. A relatively high amount of WT1 mRNA was measured by quantitative competitive reverse transcription polymerase using RNA extracted from crocidolite-induced tumors. However, WT1 seems to be expressed on a rather low level in tumors induced by B[a]P. PMID- 9400708 TI - Mechanism of asbestos-mediated DNA damage: role of heme and heme proteins. AB - Several observations, including studies from this laboratory, demonstrate that asbestos generates free radicals in the biological system that may play a role in the manifestation of asbestos-related cytotoxicity and carcinogenicity. It has also been demonstrated that iron associated with asbestos plays an important role in the asbestos-mediated generation of reactive oxygen species. Exposure to asbestos leads to degradation of heme proteins such as cytochrome P450-releasing heme in cytosol. Our simulation experiments in the presence of heme show that such asbestos-released heme may increase lipid peroxidation and can cause DNA damage. Further, heme and horseradish peroxidase (HRP) can cause extensive DNA damage in the presence of asbestos and hydrogen peroxide/organic peroxide/hydroperoxides. HRP catalyzes oxidation reactions in a manner similar to that of prostaglandin H synthetase. Iron released from asbestos is only partially responsible for DNA damage. However, our studies indicate that DNA damage mediated by asbestos in vivo may be caused by a combination of effects such as the release and participation of iron, heme, and heme moiety of prostaglandin H synthetase in free radical generation from peroxides and hydroperoxides. PMID- 9400709 TI - Asbestos, asbestosis, and lung cancer: observations in Quebec chrysotile workers. AB - One prospective epidemiologic study of asbestos cement workers with radiological small opacities has been cited as a rationale for attributing excess lung cancer to asbestosis. This approach could have considerable practical value for disease attribution in an era of decreasing exposure. However, a recent International Agency for Research on Cancer review concludes that the mechanism of production of asbestos-related lung cancer are unknown. Asbestosis, therefore, cannot be a biologically effective dose marker of lung cancer susceptibility. Asbestosis nonetheless would be useful in identifying asbestos-attributable lung cancer cases if it could be proven an infallible exposure indicator. In this study, we tested this hypothesis in the chrysotile miners and millers of Quebec, Canada. We examined exposure histories, autopsy records, and lung fiber content for 111 Quebec chrysotile miners and millers. If the hypothesis of an asbestosis requirement for lung cancer attribution were accurate, we would expect as asbestosis diagnosis to separate those with lung cancer and high levels of exposure from those with lower levels of exposure in a specific and sensitive manner. This is the first such study in which historical job-based individual estimates based on environmental measurements, lung fiber content, exposure timing, and complete pathology records including autopsies were available for review. We found significant excesses of lung tremolite and chrysotile and estimated cumulative exposure in those with lung cancer and asbestosis compared to those with lung cancer without asbestosis. However, when the latter were directly compared on a case-by-case basis, there was a marked overlap between lung cancer cases with and without asbestosis regardless of the measure of exposure. Smoking habits did not differ between lung cancer cases with and without asbestosis. In regression models, smoking pack-years discriminated between those with the without lung cancer, regardless of asbestosis status. Most seriously, the pathologic diagnosis of asbestosis itself seemed arbitrary in many cases. We conclude that although the presence of pathologically diagnosed asbestosis is a useful marker of exposure, the absence of this disease must be regarded as one of many factors in determining individual exposure status and disease causation. PMID- 9400710 TI - Cell signaling pathways elicited by asbestos. AB - In recent years, it has become apparent that minerals can trigger alterations in gene expression by initiating signaling events upstream of gene transactivation. These cascades may be initiated at the cell surface after interaction of minerals with the plasma membrane either through receptorlike mechanisms or integrins. Alternatively, signaling pathways may be stimulated by active oxygen species generated both during phagocytosis of minerals and by redox reactions on the mineral surface. At least two signaling cascades linked to activation of transcription factors, i.e., DNA-binding proteins involved in modulating gene expression and DNA replication, are stimulated after exposure of lung cells to asbestos fibers in vitro. These include nuclear factor kappa B (NF kappa B) and the mitogen-activated protein kinase (MAPK) cascade important in regulation of the transcription factor, activator protein-1 (AP-1). Both NF kappa B and AP-1 bind to specific DNA sequences within the regulatory or promoter regions of genes that are critical to cell proliferation and inflammation. Unraveling the cell signaling cascades initiated by mineral dusts and pharmacologic inhibition of these events may be important for the control and treatment of mineral-associated occupational diseases. PMID- 9400711 TI - Possible role of lipid peroxidation in the induction of NF-kappa B and AP-1 in RFL-6 cells by crocidolite asbestos: evidence following protection by vitamin E. AB - Asbestos fibers cause persistent induction of the oxidative stress sensitive transcription factors nuclear factor kappa-B (NF-kappa B) and activator protein-1 (AP-1) in mammalian cells. These transcription factors play an important role in the regulation of cellular activity. Lipid peroxidation, mediated by reactive oxygen species, is thought to be a possible mechanism in the pathogenicity of asbestos fibers. These studies were designed to determine if crocidolite asbestos induced lipid peroxidation plays a role in the mechanism of formation of NF-kappa B and AP-1. Treatment of a rat lung fibroblast cell line (RFL-6) with crocidolite asbestos in the presence and absence of the membrane antioxidant vitamin E decreased the levels of crocidolite-induced AP-1 and NF-kappa B to background levels. Preincubation of RFL-6 cells with 5,8,11,14-eicosatetraynoic acid, an inhibitor of arachidonic acid metabolism, prior to exposure to crocidolite, abrogated crocidolite-induced NF-kappa B DNA-binding activity to background levels. Coincubation with indomethacin, a cyclooxygenase inhibitor, had no effect on NF-kappa B DNA-binding activity induced by crocidolite. However, nordihydroguaiaretic acid, a lipoxygenase inhibitor, decreased levels of NF-kappa B to background levels. This would suggest that lipoxygenase metabolites of arachidonic acid, produced following lipid peroxidation, are involved in the cellular signalling events to NF-kappa B transcription factor induction by asbestos. PMID- 9400713 TI - Asbestos and silica-induced changes in human alveolar macrophage phenotype. AB - The mechanism by which fibrogenic particulates induce inflammation that can progress to lung fibrosis is uncertain. The alveolar macrophage (AM) has been implicated in the inflammatory process because of its function and reported release of inflammatory mediators when isolated from fibrotic patients. It has been recently shown that fibrogenic, but not nonfibrogenic, particulates are highly potent in inducing apoptosis of human AM. In this study, we tested the hypothesis that fibrogenic particulates could shift the phenotypic ratio of human AM to a more inflammatory condition. The macrophage phenotypes were characterized by flow cytometry targeting the RFD1 and RFD7 epitopes. Results demonstrated that chrysotile and crocidolite asbestos, as well as crystalline silica, but not titanium dioxide or wollastonite, increased the RFD1+ phenotype (inducer or immune activator macrophages) and decreased the RFD1+ RFD7+ phenotype (suppressor macrophages). These results provide a mechanistic explanation that may link apoptosis (namely, suppressor macrophages) to a shift in the ratio of macrophage phenotypes that could initiate lung inflammation. PMID- 9400712 TI - Increased focal adhesion kinase- and urokinase-type plasminogen activator receptor-associated cell signaling in endothelial cells exposed to asbestos. AB - Exposure of low-passage endothelial cells in culture to nonlethal amounts of asbestos, but not refractory ceramic fiber-1, increases cell motility and gene expression. These changes may be initiated by the fibers mimicking matrix proteins as ligands for receptors on the cell surface. In the present study, 1- to 3-hr exposures of endothelial cells to 5 mg/cm2 of chrysotile asbestos caused marked cell elongation and motility. However, little morphological change was seen when chrysotile was added to cells pretreated with either mannosamine to prevent assembly of glycophosphatidylinositol (GPI)-anchored receptors or with herbimycin A to inhibit tyrosine kinase activity. Affinity purification of GPI anchored urokinase-type plasminogen activator receptor (uPAR) from chrysotile exposed cells demonstrated that asbestos altered the profile of proteins and phosphoproteins complexed with this receptor. Tyrosine kinase activities in the complexes were also increased by asbestos. Immunoprecipitations with selective monoclonal antibodies demonstrated that both chrysotile and crocidolite asbestos increase kinase activities associated with p60 Src or p120 focal adhesion kinase (FAK). Further, chrysotile also changed the profile of proteins and phosphoproteins associated with FAK in intact cells. These data suggest that asbestos initiates endothelial cell phenotypic change through interactions with uPAR-containing complexes and that this change is mediated through tyrosine kinase cascades. PMID- 9400714 TI - p53, Cip1, and Gadd153 expression following treatment of A549 cells with natural and man-made vitreous fibers. AB - DNA damage induced by chemicals and ionizing radiation is associated with the expression of negative regulators of the cell cycle. The arrest of cells in G1 and G2 phases of the cell cycle provides time for DNA repair. Asbestos fibers are carcinogenic when inhaled by both humans and animals; however, the mechanism by which the fibers exert their effect is unknown. This work was undertaken to determine whether the expression of DNA damage-inducible genes differs between crocidolite, a fiber positive for lung tumors, and JM 100 glass microfiber, which is negative for lung tumors when inhaled by rats. Temporal and dose-related expressions of p53, Cip1, and Gadd153 proteins were determined in cultured A549 cells treated with either Union Internationale Contre le Cancer crocidolite or JM 100 for 20 hr and cultured in fresh media. Immunolabeled cells were analyzed by flow cytometry, and the increased number of protein-expressing cells was determined by subtracting the expression in unexposed cells from exposed cells. Crocidolite induced the expression of all three proteins with a maximum expression after approximately 18 hr in fresh media. At a similar time point, JM 100 did not markedly induce the three proteins. Crocidolite also induced a dose dependent increase in the number of cells in the G2 phase of the cell cycle. These results show that asbestos behaves like ionizing radiation and genotoxic chemicals by inducing proteins associated with DNA damage and cell-cycle arrest. The clear difference in response between crocidolite and JM 100 may help elucidate the mechanism of action of toxic and nontoxic fibers. PMID- 9400715 TI - Crocidolite asbestos induces apoptosis of pleural mesothelial cells: role of reactive oxygen species and poly(ADP-ribosyl) polymerase. AB - Mesothelial cells, the progenitor cells of the asbestos-induced tumor mesothelioma, are particularly sensitive to the toxic effects of asbestos, although the molecular mechanisms by which asbestos induces injury in mesothelial cells are not known. We asked whether asbestos induced apoptosis in mesothelial cells and whether reactive oxygen species were important. Rabbit pleural mesothelial cells were exposed to crocidolite asbestos or control particles (1-10 micrograms/cm2) over 24 hr and evaluated for oligonucleosomal DNA fragmentation, loss of membrane phospholipid asymmetry, and nuclear condensation. Asbestos fibers, not control particles, induced apoptosis in mesothelial cells by all assays. Induction of apoptosis was dose dependent; crocidolite (5 micrograms/cm2) induced apoptosis (15.0 +/- 1.1%, mean +/- SE; n = 12) versus control particles (< 4%), as measured by appearance of nuclear condensation. Apoptosis induced by asbestos, but not by actinomycin D, was inhibited by extracellular catalase, superoxide dismutase in the presence of catalase, hypoxia (8% oxygen), deferoxamine, and 3-aminobenzamide (an inhibitor of the nuclear enzyme, poly(adenosine diphosphate-ribosyl) polymerase). We conclude that asbestos induces apoptosis in mesothelial cells via reactive oxygen species. We speculate that escape from this pathway could allow the abnormal survival of mesothelial cells with asbestos-induced mutations. PMID- 9400716 TI - Alterations in protein glycosylation in PMA-differentiated U-937 cells exposed to mineral particles. AB - Carbohydrate moieties of cell glycoconjugates play a pivotal role in molecular recognition phenomena involved in the regulation of most biological systems and the changes observed in cell surface carbohydrates during cell activation or differentiation frequently modulate certain cell functions. Consequently, some aspects of macrophage response to particle exposure might conceivably result from alterations in glycosylation. Therefore, the effect of mineral particles on protein glycosylation was investigated in phorbol myristate acetate (PMA) differentiated U-937. Jacalin, a lectin specific for O-glycosylated structures, showed a global increase in O-glycosylation in particle-treated cells. In contrast, no significant modifications were observed with concanavalin A, a lectin that recognizes certain N-glycosylated structures. The sialic acid specific lectins Sambucus nigra agglutinin and Maackia amurensis agglutinin and the galactose-specific lectin Ricinus communis agglutinin revealed a complex pattern of alterations in glycoprotein glycosylation after crystalline silica or manganese dioxide treatments. Expression of sialyl Lewis(x), a glycosylated structure implicated in leukocyte trafficking, could not be detected in control or treated cells. This finding was consistent with the decrease in sialyl Lewis(x) expression observed during PMA-induced differentiation. In conclusion, various treatments used in this study induced quantitative as well as qualitative changes in protein glycosylation. Whether these changes are due to glycosidase release or to an alteration in glycosyltransferase expression remains to be determined. The potential functional implications of these changes are currently under investigation. PMID- 9400717 TI - Cytokines and particle-induced inflammatory cell recruitment. AB - The inflammatory response is a key component of host defense. However, excessive or persistent inflammation can contribute to the pathogenesis of disease. Inflammation is regulated, in part, by cytokines, which are small, typically glycosylated proteins that interact with membrane receptors to regulate cellular processes such as proliferation, differentiation, and secretion. During the past 10 years studies in humans and experimental animals have demonstrated that a cytokine called tumor necrosis factor alpha (TNF-alpha) plays a key role in the initiation of inflammatory responses in the lung and other tissues, including inflammation resulting from inhalation of noxious particles. There is now compelling evidence that one of the pathways by which inhaled particles stimulate the recruitment and subsequent activation of inflammatory cells is through the activation of lung macrophages to release TNF-alpha. TNF-alpha then acts via paracrine and autocrine pathways to stimulate cells to release other cytokines known as chemokines, which are directly chemotactic to leukocytes and other cells that participate in inflammatory and wound healing responses. In addition to a TNF-alpha-mediated pathway, there is growing evidence that some particles such as quartz and crocidolite can directly activate lung epithelial cells to release chemokines such as macrophage inflammatory protein-2, cytokine-induced neutrophil chemoattractant, and interleukin-8. A direct stimulatory effect of particles on lung epithelium may represent an additional or alternate pathway by which inhaled particles may elicit inflammation in the lung. Recent studies have suggested that oxidative stress may be a component of the mechanism by which particles activate cytokine production in cells such as macrophages and epithelial cells. The contribution of oxidative stress to particle-induced cytokine gene expression appears to be mediated, at least in part, through activation of the transcription factor nuclear factor kappa B. PMID- 9400718 TI - Analyzing the genes and peptide growth factors expressed in lung cells in vivo consequent to asbestos exposure and in vitro. AB - Inhalation of fibrogenic particles causes injury to the bronchiolar-alveolar epithelium. Consequently, there is a rapid proliferative response as the epithelium recovers and interstitial mesenchymal cells divide and produce connective tissue. In our model of brief (5-hr) exposure to chrysotile asbestos (approximately 1000 fibers/cc) in rats and mice, these events result in focal scarring at the bronchiolar-alveolar duct junctions in a histopathologic pattern identical to that seen in asbestos-exposed individuals. After 3 consecutive days of exposure, these lesions persist for at least 6 months postexposure. We postulate that cell proliferation and production of extracellular matrix is mediated in large part by three peptide growth factors, transforming growth factors alpha and beta (TGF-alpha and -beta), and platelet-derived growth factor (PDGF) A- and B-chains. To test this hypothesis in part, we have asked whether the genes that code for these growth factor proteins are activated at sites of asbestos-induced lung injury. If these genes were not activated, it would be reasonable to suspect that other potent growth factors and cytokines released during lung injury could be the primary mediators of fibroproliferative lung disease. In the studies reported here, we show, by in situ hybridization (ISH) and immunohistochemistry, that the four genes and their concomitant proteins are expressed within 24 hr in the bronchiolar-alveolar epithelium and underlying mesenchymal cells. RNase protection assay and ISH showed that the PDGF gene was upregulated during the first 5 hr of exposure and all the gene products remained above control levels for at least 2 weeks postexposure. TGF-alpha is a potent mitogen for epithelial cells, whereas the PDGF isoforms are potent growth factors for mesenchymal cells. TGF-beta retards fibroblast growth but stimulates extracellular matrix synthesis. Further studies using gene knockouts, appropriate antibodies, or antisense technology will be necessary to prove whether any of the growth factors are playing a significant role in fibrogenic lung disease. In addition, we have carried out a series of studies using type II alveolar epithelial cells purified from adult mouse lungs and maintained for up to 8 weeks in serum-free culture. These cells exhibit high transepithelial resistance values and they release TGF-beta 1 and -beta 2. This cell type also has been cultured from TGF-alpha knockout mice, resulting in monolayers with increased transepithelial resistance. This combination of studies in vivo and in vitro will allow us to pursue the mechanisms through which growth factors mediate lung fibrosis. PMID- 9400719 TI - Effects of mineral fibers on the expression of genes whose product may play a role in fiber pathogenesis. AB - To determine which factors are useful for the risk assessment of man-made fibers, we examined the gene expression of proinflammatory cytokines, growth factors, manganese superoxide dismutase (MnSOD), and inducible nitric oxide synthase (iNOS) in mineral fiber-exposed rats by means of reverse transcription-polymerase chain reaction (RT-PCR). Male Wistar rats received a single intratracheal instillation of either saline (control) or two types of fibers (2 mg of Union Internationale Centre le Cancer (UICC) chrysotile or alumina silicate refractory ceramic fiber [RCF]). Expression of interleukin-1 alpha (IL-1 alpha), interleukin 6 (IL-6), tumor necrosis factor alpha (TNF-alpha), platelet-deriving growth factor-A, (PDGF-A), platelet-deriving growth factor-B (PDGF-B), transforming growth factor beta 1 (TGF-beta 1), basic fibroblast growth factor (bFGF), MnSOD, and iNOS mRNA from lung and lipopolysaccharide (LPS)-stimulated alveolar macrophages (AM) were assessed by RT-PCR. Among these factors, IL-1 alpha, TNF alpha, IL-6, bFGF, and iNOS would be the possible parameters for the risk assessment of fibers. In a follow-up study, we investigated the time course (3 days, 1 week, 1 month, and 3 months) of expression of IL-1 alpha and TNF-alpha by LPS-stimulated AM exposed to mineral fibers in vivo. Male Wistar rats were instilled intratracheally with saline or fibers (2 mg of Union Internationale Contre le Cancer UICC crocidolite or potassium octatitanate whisker [TW]). The expression of IL-1 alpha mRNA by fibers was greatest in TW, crocidolite, chrysotile, and RCF-instilled rat AM, in that order. The increase of IL-1 alpha and TNF-alpha mRNA in AM peaked at 1 month and 3 days after exposure to crocidolite or TW, respectively. The expression of IL-1 alpha by fibers (crocidolite, chrysotile, TW, and RCF) may be a good indicator of the pathologic potential of fibers. PMID- 9400720 TI - Particulate-cell interactions and pulmonary cytokine expression. AB - The type II cell plays an important role in the response of the alveolar epithelium after lung injury through its synthesis and secretion of pulmonary surfactant, and by acting as the stem cell for the replacement of damaged type I epithelial cells. The nonciliated bronchiolar epithelial (Clara) cell is thought to play a similar role during repair of the bronchiolar epithelium. Recent evidence has suggested that epithelial cells may participate in aspects of the inflammatory response and regulation of fibroblast growth during pulmonary fibrosis through the production of and response to specific growth factors and cytokines. The cellular and molecular responses of epithelial cells and how they lead to the progression of events that defines the pulmonary parenchymal response to a class of particles is unclear. We used particles differing in size, chemical composition, and fibrogenicity in vivo and in vitro to elucidate early changes in proinflammatory and profibrotic cytokine and antioxidant gene expression in lung cells. Early increases in mRNA and protein for the proinflammatory cytokines interleukin (IL)-1 beta, IL-6, and tumor necrosis factor alpha have been observed in epithelial cells following exposure. These are accompanied by changes in specific epithelial genes including surfactant protein C and Clara cell secretory protein. The data indicate that effects on the epithelium are due to direct interactions with particles, not a result of macrophage-derived mediators, and suggest a more significant role in the overall pulmonary response than previously suspected. These results suggest that type II cell growth factor production may be significant in the pathogenesis of pulmonary fibrosis. PMID- 9400721 TI - Localization of intercellular adhesion molecule-1 (ICAM-1) in the lungs of silica exposed mice. AB - Intercellular adhesion molecule-1 (ICAM-1) is expressed on a variety of cells including endothelial cells, alveolar epithelial cells, and alveolar macrophages. Endothelial/epithelial cell ICAM-1 participates in the migration of leukocytes out of the blood in response to pulmonary inflammation, whereas alveolar macrophage ICAM-1 may represent cell activation. Our previous studies have shown that there is increased expression of ICAM-1 in lung tissue during acute inflammation following intratracheal injection with silica particles (2 mg/mouse). This increased expression was shown to play a role, in part, in the migration of neutrophils from the circulation into the tissue parenchyma. The aim of the current work is to localize expression of ICAM-1 during acute inflammation in lungs of mice exposed to either silica or the nuisance dust, titanium dioxide. In silica-exposed mice, a significant increase in ICAM-1 was detected on day-1 and localized by immunohistochemistry to aggregates of pulmonary macrophages and to type II epithelial cells. Areas of the lung with increased ICAM-1 expression also showed increased tumor necrosis factor alpha expression. Immunocytochemical staining of bronchoalveolar lavage (BAL) cells demonstrated increased ICAM-1 expression associated with alveolar macrophages 3, 5, and 7 days following silica exposure. Finally, soluble ICAM-1 levels in the BAL fluid were significantly increased in mice exposed to silica on the same days. Titanium dioxide exposure elicited a minimal increase in expression of ICAM-1 in the lungs. These data demonstrate that exposure to the toxic particle silica specifically increases ICAM-1 expression localized to pulmonary macrophages and type II epithelial cells. PMID- 9400722 TI - Alveolar macrophage interaction with air pollution particulates. AB - We applied flow cytometric analysis to characterize the in vitro response of alveolar macrophages (AM) to air pollution particulates. Normal hamster AM were incubated with varying concentrations of residual oil fly ash (ROFA) or concentrated ambient air particulates (CAP). We found a dose-dependent increase in AM-associated right angle light scatter (RAS) after uptake of ROFA (e.g., mean channel number 149.4 +/- 6.5, 102.5 +/- 4.1, 75.8 +/- 3.5, and 61.0 +/- 4.6 at 200, 100, 50, and 25 mg/ml, respectively) or CAP. A role for scavenger-type receptors (SR) in AM uptake of components of ROFA and CAP was identified by marked inhibition of RAS increases in AM pretreated with the specific SR inhibitor polyinosinic acid. We combined measurement of particle uptake (RAS) with flow cytometric analysis of intracellular oxidation of dichlorofluorescin. Both ROFA and CAP caused a dose-related intracellular oxidant stress within AM, comparable to that seen with phorbol myristate acetate (PMA) (e.g., fold increase over control, 6.6 +/- 0.4, 3.6 +/- 0.4, 4.6 +/- 0.5, 200 mg/ml ROFA, 100 mg/ml ROFA, and 10(-7) M PMA, respectively). We conclude that flow cytometry of RAS increases provides a useful relative measurement of AM uptake of complex particulates within ROFA and CAP. Both ROFA and CAP cause substantial intracellular oxidant stress within AM, which may contribute to subsequent cell activation and production of proinflammatory mediators. PMID- 9400724 TI - Early mesothelial cell proliferation after asbestos exposure: in vivo and in vitro studies. AB - There is some evidence that proliferation of pleural mesothelial cells (MC) occurs soon after deposition of asbestos fibers. To study this effect, we instilled a single dose of 0.1 mg crocidolite into the lungs of mice for 1 and 6 weeks and counted labeled nuclei after 3H-thymidine (3HT) injection. Short fibers (< 1 micron) induced little change in the lung; they were mostly phagocytized and had a minimal effect on MC labeling. Long fibers up to 20 microns damaged the bronchiolar epithelium and were incorporated into connective tissue. Increased 3HT uptake was seen in fibroblasts and epithelial cells and also in MC, which peaked at 2% labeled at 1 week compared to near 0% labeling in controls. No fibers were found in or near labeled MC, which suggested that a cytokine generated in the lung during the early response phase might induce MC proliferation. To look for a cytokine effect in vitro, we instilled asbestos into rat lungs and, after 1 and 6 weeks, bronchoalveolar and pleural lavage fluids as well as macrophages were collected. Alveolar macrophages contained fibers, but pleural macrophages (PM) did not. After short-term culture, macrophage supernatants and the lavage fluids were tested on rat lung MC in culture. At 1 week, PM secreted growth factor(s) for MC, and the mitogenic effect was more pronounced with lavage fluids. No effects on MC were found using material prepared 6 weeks after asbestos. The early MC growth increase was not blocked by antibodies to cytokines, such as platelet-derived growth factor, fibroblast growth factors, or tumor necrosis factor, but was inhibited by anti-keratinocyte growth factor (anti-KGF). The results show that an early growth phase of MC after asbestos exposure appears unrelated to particle translocation to the pleura but is associated with cytokine release, most likely KGF, by lung cells. PMID- 9400723 TI - Immunohistochemical localization of transforming growth factor beta isoforms in asbestos-related diseases. AB - Transforming growth factor beta (TGF-beta), a multifunctional cytokine and growth factor, plays a key role in scarring and fibrotic processes because of its ability to induce extracellular matrix proteins and modulate the growth and immune function of many cell types. These effects are important in inflammatory disorders with fibrosis and cancer. The asbestos-related diseases are characterized by fibrosis in the lower respiratory tract and pleura and increased occurrence of lung cancer and mesothelioma. We performed immunohistochemistry with isoform-specific antibodies to the three TGF-beta isoforms on 16 autopsy lungs from Quebec, Canada, asbestos miners and millers. There was increased immunolocalization of all three TGF-beta isoforms in the fibrotic lesions of asbestosis and pleural fibrosis. The hyperplastic type II pneumocytes contained all three isoforms. By contrast, there was differential spatial immunostaining for the TGF-beta isoforms in malignant mesothelioma, with TGF-beta 1 in the stroma but TGF-beta 2 in the tumor cells. These data are consistent with an important role for TGF-beta in accumulation of extracellular matrix and cell proliferation in asbestos-related diseases. PMID- 9400725 TI - Comparison of pleural responses of rats and hamsters to subchronic inhalation of refractory ceramic fibers. AB - In the present subchronic study, we compared pleural inflammation, visceral pleural collagen deposition, and visceral and parietal pleural mesothelial cell proliferation in rats and hamsters identically exposed to a kaolin-based refractory ceramic fiber, (RCF)-1 by nose-only inhalation exposure, and correlated the results to translocation of fibers to the pleural cavity. Fischer 344 rats and Syrian golden hamsters were exposed to 650 fibers/cc of RCF-1, for 4 hr/day, 5 days/week for 12 weeks. Following 4 and 12 weeks of exposure, and after a 12-week recovery period, pleural lavage fluid was analyzed for cytologic and biochemical evidence of inflammation. Visceral and parietal pleural mesothelial cell proliferation was assessed by immunocytochemical detection of bromodeoxyuridine incorporation. Pleural collagen was quantitated using morphometric analysis of lung sections stained with Sirius Red. Fiber-exposed rats and hamsters had qualitatively similar pleural inflammation at each time point. Mesothelial cell proliferation was more pronounced in hamsters than in rats at each time point and at each site. In both species, the mesothelial cell labeling index was highest in the parietal pleural mesothelial cells lining the surface of the diaphragm at each time point. Hamsters but not rats had significantly elevated collagen in the visceral pleura at the 12-week postexposure time point. Fibers were found in the pleural cavities of both species at each time point. These fibers were generally short and thin. These results suggest that mesothelial cell proliferation and fibroproliferative changes in the pleura of rodents following short-term inhalation exposure are associated with fiber translocation to the pleura and may be predictive of chronic pleural disease outcomes following long-term exposure. PMID- 9400726 TI - Mechanisms of mineral dust-induced emphysema. AB - Mineral dust exposure can result in emphysema and chronic airflow obstruction. We postulated that dust-induced emphysema has a pathogenesis similar to that in cigarette smoke-induced emphysema, namely, excess release of proteolytic enzymes from dust-evoked inflammatory cells, and inactivation of alpha-1-antitrypsin (A1AT) by dust-catalyzed formation of oxidants. To test this theory we examined the antiproteolytic activity of A1AT exposed to quartz in vitro and found that it was decreased in a dose-response fashion. Catalase prevented this effect, which suggested that it was mediated by quartz-generated hydrogen peroxide. We also showed that a variety of dusts could oxidize methionine to methionine sulfoxide in vitro, using either pure amino acid or whole protein. The relative order of activity was coal > quartz > titanium dioxide. Lastly, we used a new high performance liquid chromatography technique to demonstrate that quartz, coal, and titanium dioxide produced connective tissue breakdown in rat lungs, as determined by the appearance of desmosine and hydroxyproline in lavage fluid after dust instillation. On a particle-for-particle basis, the order of dust potency was similar to that for methionine oxidation. Connective tissue breakdown was associated with elevations of both polymorphonuclear leukocytes and macrophages in lavage fluid, and it is unclear whether one or both of these types of inflammatory cell mediates this process. These observations support our theory that dust-induced emphysema and smoke-induced emphysema occur through similar mechanisms. PMID- 9400727 TI - Lung proliferative and clearance responses to inhaled para-aramid RFP in exposed hamsters and rats: comparisons with chrysotile asbestos fibers. AB - This study compared pulmonary effects of para-aramid respirable-sized, fiber shaped particles (RFP) (p-aramid fibrils) and chrysotile asbestos fiber exposures in rats. Additional p-aramid inhalation studies were conducted in hamsters to compare species responses. The hamster results are preliminary. The parameters studied were clearance/biopersistence of inhaled p-aramid RFP or size-separated asbestos fibers as well as pulmonary cell proliferation and inflammation indices after 2-week inhalation exposures. Rats were exposed nose only to chrysotile asbestos fibers at concentrations of 459 and 782 fibers/ml or to p-aramid RFP at 419 or 772 fibrils/ml. Hamsters were exposed whole body to p-aramid RFP at concentrations of 358 and 659 fibrils/ml. Subsequently, animals were assessed immediately (time 0) as well as 5 days (10 days for hamsters), 1, 3, 6, and 12 months postexposure. Lung burdens for the p-aramid-exposed rats were 4.8 x 10(7) and 7.6 x 10(7) fibrils/lung, with similar numbers of chrysotile fibers > 5 microns recovered from the lungs of asbestos-exposed rats. In comparison, 1.4 x 10(6) fibrils/lung were recovered in the high-dose hamster group. Biopersistence studies in p-aramid-exposed rats and hamsters demonstrated an initial increase (relative to time 0) in retained p-aramid fibrils during the first month postexposure, which indicated breakage or shortening of inhaled fibrils. This result was associated with a progressive reduction, and increased residence time in the lung, in the mean lengths of the fibrils, which signified biodegradability of inhaled p-aramid fibrils in both species. In contrast, clearance of short chrysotile asbestos fibers was rapid, but clearance of the long chrysotile fibers was slow or insignificant, as evidenced by a progressive increase over time in the mean lengths of fibers recovered from the lungs of exposed rats. Two-week, high-dose exposures to p-aramid in both rats and hamsters produced transient increases in pulmonary inflammatory and cell proliferative responses. In contrast, inhalation of size-separated chrysotile asbestos fibers in rats produced persistent increases in cell labeling indices of airway, alveolar, and subpleural cells measured through a period of 1 to 3 months postexposure. These results suggest that inhaled p-aramid RFP are biodegradable in the lungs of exposed rats and hamsters. In contrast, exposures to chrysotile asbestos fibers in rats resulted in a selective pulmonary retention of long chrysotile fibers. PMID- 9400728 TI - Chronic inhalation study of fiber glass and amosite asbestos in hamsters: twelve month preliminary results. AB - The effects of chronic inhalation of glass fibers and amosite asbestos are currently under study in hamsters. The study includes 18 months of inhalation exposure followed by lifetime recovery. Syrian golden hamsters are exposed, nose only, for 6 hr/day, 5 day/week to size-selected test fibers: MMVF10a (Schuller 901 insulation glass); MMVF33 (Schuller 475 durable glass); amosite asbestos (three doses); or to filtered air (controls). Here we report interim results on airborne fiber characterization, lung fiber burden, and pathology (preliminary) through 12 months. Aerosolized test fibers averaged 15 to 20 microns in length and 0.5 to 1 micron in diameter. Target aerosol concentrations of World Health Organization (WHO) fibers (longer than 5 microns) were 250 fibers/cc for MMVF10a and MMVF33, and 25, 125, or 250 fibers/cc for amosite. WHO fiber lung burdens showed time-dependent and (for amosite) dose-dependent increases. After a 12 month exposure, lung burdens of fibers longer than 20 microns were greatest with amosite high and mid doses, similar for low-dose amosite and MMVF33, and smaller for MMVF10a. Biological responses of animals exposed for 12 months to MMVF10a were limited to nonspecific pulmonary inflammation. However, exposures to MMVF33 and each of three doses of amosite were associated with lung fibrosis and possible mesotheliomas (1 with MMVF33 and 2, 3, and 1 with amosite low, mid, and high doses, respectively). Pulmonary and pleural changes associated with amosite were qualitatively and quantitatively more severe than those associated with MMVF33. As of the 12-month time point, this study demonstrates that two different fiber glass compositions with similar fiber dimensions but different durabilities can have distinctly different effects on the hamster lung and pleura after inhalation exposure. (Preliminary tumor data through 18 months of exposure and 6 weeks of postexposure recovery became available as this manuscript went to press: No tumors were observed in the control or MMVF10a groups, and no additional tumors were observed in the MMVF33 group; however, a number of additional mesotheliomas were observed in the amosite groups. PMID- 9400729 TI - Sites of particle retention and lung tissue responses to chronically inhaled diesel exhaust and coal dust in rats and cynomolgus monkeys. AB - The usefulness of pulmonary carcinogenicity data from rats exposed to high concentrations of particles for quantitatively predicting lung cancer risk in humans exposed to much lower environmental or occupational concentrations has been questioned. The results of several chronic inhalation bioassays of poorly soluble, nonfibrous particles have suggested that rats may be more prone than other rodent species to develop persistent pulmonary epithelial hyperplasia, metaplasia, and tumors in response to the accumulation of inhaled particles. In addition, rats and primates differ in their pulmonary anatomy and rate of particle clearance from the lung. This paper reviews results of recent Lovelace Respiratory Research Institute (Albuquerque, NM) investigations that directly compared the anatomical patterns of particle retention and the lung tissue responses of rats and monkeys exposed chronically to high occupational concentrations of poorly soluble particles. Lung sections from male cynomolgus monkeys and F344 rats exposed 7 hr/day, 5 days/week for 24 months to filtered ambient air, diesel exhaust (2 mg soot/m3), coal dust (2 mg respirable particulate material/m3), or diesel exhaust and coal dust combined (1 mg soot and 1 mg respirable coal dust/m3) were obtained from a study conducted at the U.S. National Institute for Occupational Safety and Health and examined histopathologically and morphometrically. Within each species, the sites of particle retention and lung tissue responses were the same for diesel soot, coal dust, and combined material. Rats retained a significantly greater portion of the particulate material in the lumens of alveolar ducts and alveoli than monkeys. Conversely, monkeys retained a significantly greater portion of the particulate material in the interstitium than rats. Rats, but not monkeys, had significant alveolar epithelial hyperplastic, inflammatory, and septal fibrotic responses to the retained particles. These results suggest that anatomic patterns of particle retention and lung tissue reactions in rats may not be predictive of retention patterns and tissue responses in primates that inhale poorly soluble particles at concentrations representing high occupational exposures. PMID- 9400730 TI - Short-term inhalation and in vitro tests as predictors of fiber pathogenicity. AB - A wide range of fiber types was tested in two in vitro assays: toxicity to A549 epithelial cells, as detachment from substrate, and the production of the proinflammatory cytokine tumor necrosis factor (TNF) by rat alveolar macrophages. Three of the fibers were also studied in vivo, using short-term inhalation followed by a) bronchoalveolar lavage to assess the inflammatory response and b) measurement of cell proliferation in terminal bronchioles and alveolar ducts, using incorporation of bromodeoxyuridine (BrdU). The amount of TNF produced by macrophages in vitro depended on the fiber type, with the man-made vitreous fibers, and refractory ceramic fibers being least stimulatory and silicon carbide (SiC) whiskers providing the greatest stimulation. In the epithelial detachment assay there were dose-dependent differences in the toxicity of the various fibers, with long amosite being the most toxic. However, there was no clear relationship to known chronic pathogenicity. Fibers studied by short-term inhalation produced some inflammation, but there was no clear discrimination between the responses to code 100/475 glass fibers and the more pathogenic amosite and SiC. However, measurements of BrdU uptake into lung cells showed that amosite and SiC produced a greater reaction than code 100/475, which itself caused no more proliferation than that seen in untreated lungs. These results mirror the pathogenicity ranking of the fibers in long-term experiments. In conclusion, the only test to show potential as a predictive measure of pathogenicity was that of cell proliferation in lungs after brief inhalation exposure (BrdU assay). We believe that this assay should be validated with a wider range of fibers, doses, and time points. PMID- 9400731 TI - Silica-induced apoptosis in alveolar and granulomatous cells in vivo. AB - Silica is a toxicant that can stimulate cells to produce various cellular products such as free radicals, cytokines, and growth factors. Silica and its induced substances may induce apoptosis to regulate the evolution of silica induced inflammation and fibrosis. To examine this hypothesis, groups of Wistar male rats were intratracheally instilled with different doses of Min-U-Sil 5 silica (Silica, Berkeley Springs, WV). Ten days after the instillation, we obtained cells by bronchoalveolar lavage and placed them on slides by cytospin preparation. The slides were stained with Diff-Quik (Lab Aids, Sydney, NSW, Australia) and examined under oil immersion. A substantial number of cells with apoptotic features were identified in all silica-instilled rats and the apoptosis was confirmed by agarose gel electrophoresis. The number of apoptotic cells was clearly related to silica dosage. Engulfment of apoptotic cells by macrophages was also noted. Neutrophil influx in silica-instilled rats could be saturated with the increase of silica dosage and the number of macrophages in different dose groups changed in parallel with the proportion of apoptotic cells. Fifty-six days after instillation, morphologically apoptotic cells could be identified in granulomatous cells of lung tissue from silica-instilled rats. We conclude that intratracheal instillation of silica could induce apoptosis in both alveolar and granulomatous cells, and the apoptotic change and subsequent engulfment by macrophages might play a role in the evolution of silica-induced effects. PMID- 9400732 TI - Expression of matrix metalloproteinases, tissue inhibitors of metalloproteinases, and extracellular matrix mRNA following exposure to mineral fibers and cigarette smoke in vivo. AB - To determine the effect of mineral fibers and cigarette smoke on remodeling of lung tissues, we examined matrix metalloproteinase-1 (MMP-1), MMP-2, tissue inhibitors of metalloproteinase-1 (TIMP-1), TIMP-2, and types I and IV collagen mRNA levels from rat lungs exposed to mineral fibers and/or cigarette smoke in vivo. Male Wistar rats (10 weeks of age) were given a single intratracheal instillation of 2 mg of chrysotile or alumina silicate ceramic fibers (RCF). Animals were then exposed to cigarette smoke (side stream) 5 days per week for 4 weeks. Transcriptional levels of mRNA extracted from the lungs were assessed by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR). Exposure to cigarette smoke induced increases in MMP-1 and TIMP-1 mRNA levels and decreased TIMP-2 and type I collagen mRNA levels in lung. Chrysotile or RCF stimulated the expression of MMP-1 mRNA in the lung. The mineral fibers and cigarette smoke had more than additive effects on the expression of MMP-2 and TIMP-1 in the lung. These data suggest that the imbalance of the expression of MMPs, TIMPs, and extracellular matrix may be associated with the remodeling of lung tissues induced by mineral fibers and/or cigarette smoke. PMID- 9400733 TI - Dose-response relationship of fibrous dusts in intraperitoneal studies. AB - The relationship between the number of fibers injected intraperitoneally and the occurrence of peritoneal mesotheliomas in rats was investigated using data from a series of carcinogenicity studies with several fibrous dusts. Based on observed tumor incidences ranging between 10 and 90%, the hypothesis of a common slope of dose-response relationships (parallel probit lines in probit analysis) cannot be rejected. In general, parallelism of probit lines is considered an indication of a common mode of action. Analysis of the shape of the dose-response relationship, with one apparent exception, shows virtually linear or superlinear behavior, i.e., from these data, there is no indication of a decrease in carcinogenic potency of an elementary carcinogenic unit at lower doses. PMID- 9400735 TI - Cell size of alveolar macrophages: an interspecies comparison. AB - Alveolar macrophages (AM) play a critical role in the removal of inhaled particles or fibers from the lung. Species differences in AM size may affect the number and size range of particles/fibers that can be actually phagocytized and cleared by AM. The purpose of this study was to compare the cell size of rat, hamster, monkey, and human AM by selective flow cytometric analysis of cell volume. Resident AM from CD rats, Syrian golden hamsters, cynomolgus monkeys, and nonsmoking, healthy human volunteers were harvested by standard bronchoalveolar lavage procedures. Morphometric analysis of AM was performed using a flow cytometer that generates volume signals based on the Coulter-type measurement of electrical resistance. We found that hamster and rat AM had diameters of 13.6 +/- 0.4 microns (n = 8) and 13.1 +/- 0.2 microns (n = 12), respectively. Comparatively, the AM from monkeys (15.3 +/- 0.5 microns, n = 7) and human volunteers (21.2 +/- 0.3 microns, n = 10) were larger than those from rats and hamsters. The AM from humans were significantly larger (p < 0.05) than those from all other species studied, corresponding to a 4-fold larger cell volume of human AM (4990 +/- 174 microns 3) compared to hamster (1328 +/- 123 microns 3) and rat (1166 +/- 42 microns 3) AM. In summary, we have found marked species differences in the cell size of AM. We suggest that the number and size range of particles/fibers that can be phagocytized and cleared by AM may differ among species due to inherent or acquired species differences in AM cell size. PMID- 9400734 TI - Pleural macrophage recruitment and activation in asbestos-induced pleural injury. AB - The pathogenesis of asbestos-induced pleural fibrosis is poorly understood. Moreover, there has been a long-standing controversy regarding the relative potential of different commercial types of asbestos to cause pleural disease. We postulated that inhaled asbestos fibers translocate to the pleural space where they stimulate the recruitment and activation of pleural macrophages. To test this hypothesis, and to determine whether there are differences between inhaled amphibole and serpentine asbestos, Fischer 344 rats were exposed by intermittent inhalation (6 hr/day for 5 days/week over 2 weeks) to either National Institute of Environmental Health Sciences (NIEHS) crocidolite (average concentration 7.55 mg/m3) or NIEHS chrysotile fibers (average concentration 8.51 mg/m3). Comparisons were made with sham-exposed rats. The rats were sacrificed at 1 and 6 weeks after the cessation of exposure. More pleural macrophages were recovered at 1 and 6 weeks after crocidolite and chrysotile exposure than after sham exposure. Small numbers of crocidolite fibers (approximately 1 per 4000 cells) were detected in the pleural cell pellet of one crocidolite-exposed rat by scanning electron microscopy. Pleural macrophage supernatants were assayed for production of nitric oxide (NO) (by the Griess reaction) and tumor necrosis factor alpha (TNF-alpha) (by an enzyme-linked immunosorbent assay method). Significantly greater amounts of NO as well as TNF-alpha were generated by pleural macrophages at 1 and 6 weeks after either crocidolite or chrysotile inhalation than after sham exposure. Conceivably, translocation of asbestos fibers to the pleural space may provide a stimulus for persistent pleural space inflammation, cytokine production, and the generation of toxic oxygen and nitrogen radicals. Enhanced cytokine secretion within the pleural space may in turn upregulate adhesion molecule expression and the synthesis of extracellular matrix constituents by pleural mesothelial cells. Thus, our findings may have significance for the development of asbestos-induced pleural injury. PMID- 9400736 TI - Intratracheal instillation versus intratracheal inhalation: influence of cytokines on inflammatory response. AB - Our laboratory has developed a method of particle exposure whereby anesthetized rats intratracheally inhale, at a regulated breathing rate and pressure, an aerosolized test material. This method is capable of delivering considerable doses in a short time period and, unlike the commonly used method of intratracheal instillation, does so with an even particle distribution throughout the lung. Early studies comparing the response of male Fischer 344 rats exposed to TiO2 particles of two differing primary particle sizes showed that at similar particle doses animals exposed by the two methods showed differences in response, as measured by bronchoalveolar lavage (BAL) parameters. Building on this, we sought to study the roles that macrophage inflammatory protein-2 (MIP-2) and tumor necrosis factor alpha (TNF-alpha), two cytokines thought to have proinflammatory roles in the lung, may play in the differences observed. Increases in MIP-2 protein levels in the lavaged cells, but not the supernatant, were observed in those groups where increased polymorphonuclear cells (PMN) in the lung lavage were found, but not in those where no increase in PMN levels was observed. BAL TNF-alpha levels, measured by enzyme-linked immunosorbent assay, showed no apparent correlation with cellular or biochemical BAL parameters for either particle size or dosing method. Increases in immunocytochemical staining for TNF-alpha, compared to unexposed controls, were observed in several particle exposed groups. Thus, it appears that increased BAL MIP-2 protein levels, but not TNF-alpha, correlate well with the inflammatory response, as measured by PMN numbers in lavaged cells, for both exposure systems. PMID- 9400737 TI - Efflux of reduced glutathione after exposure of human lung epithelial cells to crocidolite asbestos. AB - This study investigated glutathione (GSH) homeostasis in human lung epithelial cells (A549) exposed to crocidolite. Exposure of A549 cells to 3 micrograms/cm2 crocidolite resulted in a decrease in intracellular reduced glutathione by 36% without a corresponding increase in GSH disulfide. After a 24-hr exposure to crocidolite, 75% of the intracellular GSH lost was recovered in the extracellular medium, of which 50% was in reduced form. Since the half-life of reduced GSH in culture medium was less than 1 hr, this suggests that reduced GSH was released continuously from the cells after treatment. The release of GSH did not appear to result from nonspecific membrane damage, as there was no concomitant release of lactate dehydrogenase or 14C-adenine from loaded cells after crocidolite treatment for 24 hr. Crocidolite exposure resulted in the formation of S nitrosothiols but no increase in the level of GSH-protein mixed disulfides or GSH conjugates. Exposure of A549 cells to crocidolite for 24 hr decreased gamma glutamylcysteine synthetase (gamma-GCS) activity by 47% without changes in the activities of GSH reductase, GSH peroxidase, GSH S-transferase, or glucose-6 phosphate dehydrogenase. Treatment of cells with crocidolite pretreated with the iron chelator desferrioxamine B resulted in the same level of intracellular GSH depletion and efflux and the same decrease in gamma-GCS activity as treatment with unmodified crocidolite, which suggests that iron-catalyzed reactions were not responsible for the GSH depletion. PMID- 9400738 TI - In vivo and in vitro proinflammatory effects of particulate air pollution (PM10). AB - Epidemiologic studies have reported associations between fine particulate air pollution, especially particles less than 10 mm in diameter (PM10), and the development of exacerbations of asthma and chronic obstructive pulmonary disease. However, the mechanism is unknown. We tested our hypothesis that PM10 induces oxidant stress, causing inflammation and injury to airway epithelium. We assessed the effects of intratracheal instillation of PM10 in rat lungs. The influx of inflammatory cells was measured in bronchoalveolar lavage (BAL). Airspace epithelial permeability was assessed as total protein in bronchoalveolar lavage fluid (BALF) in vivo. The oxidant properties of PM10 were determined by their ability to cause changes in reduced glutathione (GSH) and oxidized glutathione (GSSG). We also compared the effects of PM10 with those of fine (CB) and ultrafine (ufCB) carbon black particles. Six hours after intratracheal instillation of PM10, we noted an influx of neutrophils (up to 15% of total BAL cells) in the alveolar space, increased epithelial permeability, an increase in total protein in BALF from 0.39 +/- 0.01 to 0.62 +/- 0.01 mg/ml (mean +/- SEM) and increased lactate dehydrogenase concentrations in BALF. An even greater inflammatory response was observed after intratracheal instillation of ufCB, but not after CB instillation. PM10 had oxidant activity in vivo, as shown by decreased GSH in BALF (from 0.36 +/- 0.05 to 0.25 +/- 0.01 nmol/ml) after instillation. BAL leukocytes from rats treated with PM10 produced greater amounts of nitric oxide, measured as nitrite (control 3.07 +/- 0.33, treated 4.45 +/- 0.23 mM/1 x 10(6) cells) and tumor necrosis factor alpha (control 21.0 +/- 3.1, treated 179.2 +/- 29.4 unit/1 x 10(6) cells) in culture than BAL leukocytes obtained from control animals. These studies provide evidence that PM10 has free radical activity and causes lung inflammation and epithelial injury. These data support our hypothesis concerning the mechanism for the adverse effects of particulate air pollution on patients with airway diseases. PMID- 9400739 TI - Free radical activity of PM10: iron-mediated generation of hydroxyl radicals. AB - The purpose of this study was to test the hypothesis that particulate matter < or = 10 microns in aerodynamic diameter (PM10) particles have the ability to generate free radical activity at their surface. We collected PM10 filters from the Edinburgh, United Kingdom, Enhanced Urban Network sampling site, removed particles from the filter, and tested their ability to cause free radical damage to supercoiled plasmid DNA. We found that the PM10 particles did cause damage to the DNA that was mediated by hydroxyl radicals, as shown by inhibition of the injury with mannitol. The PM10-associated hydroxyl radical activity was confirmed using a high-performance liquid chromatography-based assay to measure the hydroxyl radical adduct of salicylic acid. Desferrioxamine abolished the hydroxyl radical-mediated injury, which suggests that iron was involved. Analysis of PM10 filters confirmed the presence of large amounts of iron and leaching studies confirmed that the PM10 samples could release substantial amounts of Fe(III) and lesser amounts of Fe(II). To investigate the size of the particles involved in the hydroxyl radical injury, we centrifuged the suspension of PM10 to clarity, tested the clear supernatant, and found that it had all of the suspension activity. We conclude, therefore, that the free radical activity is derived either from a fraction that is not centrifugeable on a bench centrifuge, or that the radical generating system is released into solution. PMID- 9400740 TI - Formation and persistence of 8-oxoguanine in rat lung cells as an important determinant for tumor formation following particle exposure. AB - Exposure of rats to quartz (or various other particles) can lead to the development of lung tumors. At the moment, the mechanisms involved in particle induced tumor formation are not clarified. However, it is suggested that inflammation, in conjunction with the production of reactive oxygen species (ROS) and an enhancement of epithelial cell proliferation, may play a key role in the development of lung tumors. ROS induces 8-oxoguanine (8-oxoGua) and other mutagenic DNA oxidation products, which can be converted to mutations in proliferating cells. Mutation formation in cancer-related genes is a critical event with respect to tumor formation. In this study we investigated the effects of quartz (DQ12) and of the nontumorigenic dust corundum on the induction of 8 oxoGua in the DNA of rat lung cells, as well as on cell proliferation and pulmonary inflammation. Wistar rats were exposed by intratracheal instillation to quartz (2.5 mg/rat) or corundum (2.5 mg/rat) suspended in physiological saline; control animals exposed to physiological saline or left untreated. Measurements were carried out 7, 21, and 90 days after the exposures. 8-oxoGua levels were determined in lung tissue sections at the single cell level by immunocytological assay using a rabbit anti-8-oxoGua antibody. After exposure to quartz, 8-oxoGua levels were significantly increased at all time points of investigation. Additionally, we observed inflammation and an enhanced cell proliferation. Exposure to corundum had no adverse effects on the lung; neither increased 8 oxoGua levels nor enhanced cell proliferation or inflammation were detected. These observations support the suggestion that inflammation associated with increased 8-oxoGua levels in lung cells and increased cell proliferation is an important determinant for particle-induced development of lung tumors in the rat. PMID- 9400741 TI - Expression of inducible nitric oxide synthase and formation of nitric oxide by alveolar macrophages: an interspecies comparison. AB - Nitric oxide (NO) is suggested to play a role in mediating pulmonary injury. However, interspecies differences appear to exist in the ability of alveolar macrophages (AM) to express the inducible nitric oxide synthase (iNOS) and to generate NO. The purpose of this study was to compare iNOS expression and NO production by rat, hamster, monkey, and human AM using the identical experimental conditions in vitro. As AM donors, CD rats, Syrian golden hamsters, cynomolgus monkeys, and nonsmoking, healthy human volunteers were used. The AM were obtained by bronchoalveolar lavage and stimulated in vitro with various concentrations and combinations of lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma). The oxidation product of NO, nitrite, was measured in the AM supernatant by the Griess reaction. The expression of iNOS in AM was detected using immunocytochemistry and immunoblotting. The expression of iNOS mRNA was assessed by reverse transcriptase-polymerase chain reaction (RT-PCR). Rat AM, stimulated with either LPS or IFN-gamma, produced nitrite in a time- and dose-dependent manner. Combination of LPS and IFN-gamma resulted in a significantly enhanced nitrite formation. However, none of the treatments was able to induce hamster, monkey, or human AM to release measurable amounts of nitrite. Whereas expression of iNOS protein was only detected in stimulated rat AM, expression of iNOS mRNA was found in unstimulated and stimulated rat AM, slightly in stimulated hamster AM, but not in monkey and human AM. In conclusion, our findings point to distinct regulatory mechanisms of the NO pathway in AM from these four different species. PMID- 9400742 TI - The role of reactive oxygen and nitrogen species in the response of airway epithelium to particulates. AB - Epidemiologic and occupational studies indicate adverse health effects due to inhalation of particulate air pollutants, but precise biologic mechanisms responsible have yet to be fully established. The tracheobronchial epithelium forms the body's first physiologic barrier to such airborne pollutants, where ciliary movement functions to remove the offending substances caught in the overlying mucus layer. Resident and infiltrating phagocytic cells also function in this removal process. In this paper, we examine the role of reactive oxygen and nitrogen species (ROS/RNS) in the response of airway epithelium to particulates. Some particulates themselves can generate ROS, as can the epithelial cells, in response to appropriate stimulation. In addition, resident macrophages in the airways and the alveolar spaces can release ROS/RNS after phagocytosis of inhaled particles. These macrophages also release large amounts of tumor necrosis factor alpha (TNF-alpha), a cytokine that can generate responses within the airway epithelium dependent upon intracellular generation of ROS/RNS. As a result, signal transduction pathways are set in motion that may contribute to inflammation and other pathobiology in the airway. Such effects include increased expression of intercellular adhesion molecule 1, interleukin-6, cytosolic and inducible nitric oxide synthase, manganese superoxide dismutase, cytosolic phospholipase A2, and hypersecretion of mucus. Ultimately, ROS/RNS may play a role in the global response of the airway epithelium to particulate pollutants via activation of kinases and transcription factors common to many response genes. Thus, defense mechanisms involved in responding to offending particulates may result in a complex cascade of events that can contribute to airway pathology. PMID- 9400743 TI - Interspecies comparison of rat and hamster alveolar macrophage antioxidative and oxidative capacity. AB - Generation of oxidants has been implicated in lung injury and disease caused by a variety of inhaled agents such as ozone, particles, and mineral fibers. Antioxidants in the pulmonary system presumably provide the initial defense against such oxidants. We designed the present study to assess the oxidative and antioxidative capacity of alveolar macrophages (AM) from rats and hamsters. These two laboratory animal species commonly used in biomedical research are well known for their disparate response to pulmonary irritants/toxicants. AM from CD rats and Syrian golden hamsters were obtained by bronchoalveolar lavage. We assessed AM antioxidant levels by measuring the catalase and superoxide dismutase (SOD) activity and the intracellular concentrations of total glutathione, ascorbic acid, and alpha-tocopherol. We determined the AM oxidative capacity by assessing the ability of AM to oxidize extracellular glutathione (GSH) and to release superoxide anions. There were no significant differences in the intracellular antioxidant levels, except for catalase activity that was significantly (p < 0.05) higher in hamster AM than in rat AM. However, AM oxidative capacity was markedly different between the two species studied. The amount of spontaneous and phorbol myristate acetate (PMA)-induced GSH oxidation was about 5-fold higher in rat AM than in hamster AM, whereas the PMA-induced superoxide anion release did not differ significantly between the two rodents. In summary, our data suggest that species variation exists between the oxidative capacity of rat and that of hamster AM. Whereas the oxidative capacity of hamster AM appears to be based mainly on the formation of reactive oxygen species, it is suggested that rat AM possess an additional oxidative system. PMID- 9400744 TI - Free radical activity of industrial fibers: role of iron in oxidative stress and activation of transcription factors. AB - We studied asbestos, vitreous fiber (MMVF10), and refractory ceramic fiber (RCF1) from the Thermal Insulation Manufacturers' Association fiber repository regarding the following: free radical damage to plasmid DNA, iron release, ability to deplete glutathione (GSH), and activate redox-sensitive transcription factors in macrophages. Asbestos had much more free radical activity than any of the man made vitreous fibers. More Fe3+ was released than Fe2+ and more of both was released at pH 4.5 than at pH 7.2. Release of iron from the different fibers was generally not a good correlate of ability to cause free radical injury to the plasmid DNA. All fiber types caused some degree of oxidative stress, as revealed by depletion of intracellular GSH. Amosite asbestos upregulated nuclear binding of activator protein 1 transcription factor to a greater level than MMVF10 and RCF1; long-fiber amosite was the only fiber to enhance activation of the transcription factor nuclear factor kappa B (NF kappa B). The use of cysteine methyl ester and buthionine sulfoximine to modulate GSH suggested that GSH homeostasis was important in leading to activation of transcription factors. We conclude that the intrinsic free radical activity is the major determinant of transcription factor activation and therefore gene expression in alveolar macrophages. Although this was not related to iron release or ability to deplete macrophage GSH at 4 hr, GSH does play a role in activation of NF kappa B. PMID- 9400745 TI - Augmentation of pulmonary reactions to quartz inhalation by trace amounts of iron containing particles. AB - Fracturing quartz produces silica-based radicals on the fracture planes and generates hydroxyl radicals (.OH) in aqueous media. .OH production has been shown to be directly associated with quartz-induced cell damage and phagocyte activation in vitro. This .OH production in vitro is inhibited by desferrioxamine mesylate, an Fe chelator, indicating involvement of a Fenton-like reaction. Our objective was to determine if Fe contamination increased the ability of inhaled quartz to cause inflammation and lung injury. Male Fischer 344 rats were exposed 5 hr/day for 10 days to filtered air, 20 mg/m3 freshly milled quartz (57 ppm Fe), or 20 mg/m3 freshly milled quartz contaminated with Fe (430 ppm Fe). High Fe contamination of quartz produced approximately 57% more reactive species in water than quartz with low Fe contamination. Compared to inhalation of quartz with low Fe contamination, high Fe contamination of quartz resulted in increases in the following responses: leukocyte recruitment (537%), lavageable red blood cells (157%), macrophage production of oxygen radicals measured by electron spin resonance or chemiluminescence (32 or 90%, respectively), nitric oxide production by macrophages (71%), and lipid peroxidation of lung tissue (38%). These results suggest that inhalation of freshly fractured quartz contaminated with trace levels of Fe may be more pathogenic than inhalation of quartz alone. PMID- 9400746 TI - Involvement of protein kinase C, phospholipase C, and protein tyrosine kinase pathways in oxygen radical generation by asbestos-stimulated alveolar macrophage. AB - Although asbestos stimulates oxygen radical generation in alveolar macrophages, the exact mechanism is still not clear. The purpose of this study was to compare the ability of three asbestos fibers (amosite, chrysotile, and crocidolite) to generate oxygen radicals in macrophages and examine the mechanism of this action. All asbestos fibers were able to induce chemiluminescence but chrysotile induced maximal chemiluminescence at higher concentrations than amosite and crocidolite. Protein kinase C (PKC) inhibitors (sphingosine and staurosporine) suppressed the ability of asbestos to induce oxygen radical generation. Phospholipase C (PLC) inhibitors (U73122 and neomycin) and protein tyrosine kinase (PTK) inhibitors (erbstatin and genistein) decreased oxygen radical generation of asbestos stimulated alveolar macrophages. Oxygen radical generation was not suppressed by an adenylate cyclase activator (forskolin), a protein kinase A inhibitor (H-8), and a protein serine-threonine phosphatase inhibitor (okadaic acid). PLC and PTK inhibitors suppressed the increment of phosphoinositide turnover by amosite. These results suggest that asbestos fibers induce the generation of oxygen radicals through PTK, PLC, and PKC pathways in a dose-response pattern. PMID- 9400747 TI - Approaches to characterizing human health risks of exposure to fibers. AB - Naturally occurring and man-made (synthetic) fibers of respirable sizes are substances that have been identified by the U.S. Environmental Protection Agency (U.S. EPA) as priority substances for risk reduction and pollution prevention under the Toxic Substances Control Act (TSCA). The health concern for respirable fibers is based on the link of occupational asbestos exposure and environmental erionite fiber exposure to the development of chronic respiratory diseases, including interstitial lung fibrosis, lung cancer, and mesothelioma in humans. There is also considerable laboratory evidence indicating that a variety of fibers of varying physical and chemical characteristics can elicit fibrogenic and carcinogenic effects in animals under certain exposure conditions. This paper discusses key scientific issues and major default assumptions and uncertainties pertaining to the risk assessment of inhaled fibers. This is followed by a description of the types of assessment performed by the U.S. EPA to support risk management actions of new fibers and existing fibers under TSCA. The scope and depth of these risk assessments, however, vary greatly depending on whether the substance under review is an existing or a new fiber, the purpose of the assessment, the availability of data, time, and resources, and the intended nature of regulatory action. In general, these risk assessments are of considerable uncertainty because health hazard and human exposure information is often incomplete for most fibers. Furthermore, how fibers cause diseases and what specific determinants are critical to fiber-induced toxicity and carcinogenicity are still not completely understood. Further research to improve our knowledge base in fiber toxicology and additional toxicity and exposure data gathering are needed to more accurately characterize the health risks of inhaled fibers. PMID- 9400748 TI - Relevance of particle-induced rat lung tumors for assessing lung carcinogenic hazard and human lung cancer risk. AB - Rats and other rodents are exposed by inhalation to identify agents that might present hazards for lung cancer in humans exposed by inhalation. In some cases, the results are used in attempts to develop quantitative estimates of human lung cancer risk. This report reviews evidence for the usefulness of the rat for evaluation of lung cancer hazards from inhaled particles. With the exception of nickel sulfate, particulate agents thought to be human lung carcinogens cause lung tumors in rats exposed by inhalation. The rat is more sensitive to carcinogenesis from nonfibrous particles than mice or Syrian hamsters, which have both produced false negatives. However, rats differ from mice and nonhuman primates in both the pattern of particle retention in the lung and alveolar epithelial hyperplastic responses to chronic particle exposure. Present evidence warrants caution in extrapolation from the lung tumor response of rats to inhaled particles to human lung cancer hazard, and there is considerable uncertainty in estimating unit risks for humans from rat data. It seems appropriate to continue using rats in inhalation carcinogenesis assays of inhaled particles, but the upper limit of exposure concentrations must be set carefully to avoid false positive results. A positive finding in both rats and mice would give greater confidence that an agent presents a carcinogenic hazard to man, and both rats and mice should be used if the agent is a gas or vapor. There is little justification for including Syrian hamsters in assays of the intrapulmonary carcinogenicity of inhaled agents. PMID- 9400749 TI - Pulmonary carcinogenicity of inhaled particles and the maximum tolerated dose. AB - Chronic inhalation bioassays in rodents are used to assess pulmonary carcinogenicity for purposes of hazard identification and potentially for risk characterization. The influence of high experimental doses on tumor development has been recognized for some time and has led to the concept of maximum tolerated dose (MTD) for dose selection, with the highest dose being at the MTD. Exposure at the MTD should ensure that the animals are sufficiently challenged while at the same time the animal's normal longevity is not altered from effects other than carcinogenicity. A characteristic of exposure-dose-response relationships for chronically inhaled particles is that lung tumors are significantly increased only at high exposure levels, and that lung tumors are seen in rats only but not in mice or hamsters. This lung tumor response in rats is thought to be secondary to persistent alveolar inflammation, indicating that the MTD may have been exceeded. Thus, mechanisms of toxicity and carcinogenicity may be dose dependent and may not operate at lower doses that humans normally experience. Despite awareness of this problem, carcinogenicity bioassays that evaluate particulate compounds in rodents have not always been designed with the MTD concept in mind. This is due to several problems associated with determining an appropriate MTD for particle inhalation studies. One requirement for the MTD is that some toxicity should be observed. However, it is difficult to define what degree of toxic response is indicative of the MTD. For particle inhalation studies, various noncancer end points in addition to mortality and body weight gain have been considered as indicators of the MTD, i.e., pulmonary inflammation, increased epithelial cell proliferation, increased lung weight, impairment of particle clearance function, and significant histopathological findings at the end of a subchronic study. However, there is no general agreement about quantification of these end points to define the MTD. To determine whether pulmonary responses are indicative of the MTD, we suggest defining an MTD based on results of a multidose subchronic and chronic inhalation study with a known human particulate carcinogen, e.g., asbestos or crystalline silica. Quantification of effects in such a study using the noncancer end points listed above would identify a dose level without significant signs of toxicity at the end of the subchronic study. If this dose level still results in significant lung tumor incidence at the end of the chronic study. We will have a sound basis for characterizing the MTD and justifying its use in future particle inhalation studies. Also, a better understanding of cellular and molecular mechanisms of particle-induced lung tumors is needed to support the MTD concept. PMID- 9400750 TI - Strategies for setting occupational exposure limits for particles. AB - To set occupational exposure limits (OELs) for aerosol particles, dusts, or chemicals, one has to evaluate whether mechanistic considerations permit identification of a no observed effect level (NOEL). In the case of carcinogenic effects, this can be assumed if no genotoxicity is involved, and exposure is considered safe if it does not exceed the NOEL. If tumor induction is associated with genotoxicity, any exposure is considered to be of risk, although a NOEL may be identified in the animal or human exposure studies. This must also be assumed when no information on the carcinogenic mechanism, including genotoxicity, is available. Aerosol particles, especially fibrous dusts, which include man-made mineral fiber(s) (MMMF), present a challenge for toxicological evaluation. Many MMMF that have been investigated have induced tumors in animals and genotoxicity in vitro. Since these effects have been associated with long-thin fiber geometry and high durability in vivo, all fibers meeting such criteria are considered carcinogenic unless the opposite has been demonstrated. This approach is practicable. Investigations on fiber tumorigenicity/genotoxicity should include information on dose response, pathobiochemistry, particle clearance, and persistence of the material in the target organ. Such information will introduce quantitative aspects into the qualitative approach that has so far been used to classify fibrous dusts as carcinogens. The rationales for classifying the potential carcinogenicity of MMMF and for setting OELs used by the different European committees and regulatory agencies are described. PMID- 9400752 TI - Laser photocoagulation for central serous retinopathy. PMID- 9400751 TI - Use of mechanistic data in assessing human risks from exposure to particles. AB - The ultimate goal of toxicologic investigations of both natural and man-made fibrous and nonfibrous particles is to provide essential input for the assessment of potential human risks from exposure to these materials. The development of risk assessment procedures for airborne particles has evolved over the years. The earliest assessments for naturally occurring materials used direct human observations and incorporated safety factors to arrive at allowable human exposures. More recently, there has been a need to assess the potential risk associated with production and use of certain man-made materials for which human data are not available or are inadequate. For these materials, it has been necessary to assess human risks using data obtained from studies conducted in laboratory animals and with cells or tissues. During the last several decades, it has been suggested that data on the mechanisms by which particles cause disease could be used to reduce the uncertainty in estimates of human risks of particle exposures. This article provides comments on the use of mechanistic data in the risk assessment process and suggestions for increasing the successful development and use of mechanistic data in risk assessments conducted in the future. PMID- 9400753 TI - Complications of hydroxyapatite implants. PMID- 9400754 TI - Annular peripheral choroidal detachment after glaucoma surgery. PMID- 9400755 TI - Reappraising the risk and benefits of aggressive glaucoma therapy. PMID- 9400756 TI - Incidence and progression of cortical and posterior subcapsular opacities: the Longitudinal Study of Cataract. The LSC Group. AB - OBJECTIVE: The purpose of the study is to estimate incidence and progression rates of cortical and posterior subcapsular (PSC) opacities in the Longitudinal Study of Cataract (LSC). DESIGN: An epidemiologic study of the natural history of lens opacities in a clinic-based population. PARTICIPANTS: The LSC was based on 764 participants in an earlier case-control study of lens opacities. MAIN OUTCOME MEASURES: Baseline data, collected until 1988, included color slit and retroillumination photographs. The same data were collected at follow-up visits from 1989 to 1993. The Lens Opacities Classification System III (LOCS III) was used to assess lens changes between baseline and follow-up photographs. The product-limit method was used to estimate the incidence and progression rates. RESULTS: After 5 years of follow-up, the incidence rates for developing cortical and PSC opacities were 7.7% and 4.3%, respectively. The progression rate of pre existing cortical opacities was 16.2% after 5 years, and was twice as high as the incidence rate. The progression of pre-existing PSC opacities was much higher, and reached 55.1% after 5 years of follow-up. The incidence of newly developed cortical or PSC opacities increased with age. The incidence of PSC opacities also increased when coexisting opacities were present at baseline. CONCLUSIONS: After 5 years, 1 in every 13 patients developed new cortical opacities, and 1 in 24 developed new PSC opacities. The 5-year progression rates for cortical and PSC opacities were much higher than the incidence rates. These results can be used to estimate the rate of cortical and PSC changes in similar populations. PMID- 9400757 TI - Clinical findings and hemodynamic changes associated with severe occlusive carotid artery disease. AB - OBJECTIVE: The purpose of the study was to evaluate the ophthalmologic findings and to analyze the retrobulbar hemodynamics of patients with severe (greater than 70% stenosis) occlusive carotid artery disease (OCAD) by means of color Doppler imaging (CDI). DESIGN: A case-controlled study. PARTICIPANTS: Fifty-six consecutive patients with severe OCAD and an age- and sex-matched control group consisting of 56 healthy patients without OCAD were studied. INTERVENTION: All 112 patients underwent a complete ophthalmologic examination. Color Doppler imaging of both orbits was performed by one masked investigator. MAIN OUTCOME MEASURES: Peak systolic velocity, end diastolic velocity, and the resistive index of the ophthalmic, central retinal, and temporal short posterior ciliary arteries were measured. The authors compared the hemodynamic parameters measured in patients with severe OCAD with those obtained in the control group. The hemodynamic parameters of patients with asymmetric OCAD (stenosis > 70% in one internal carotid artery and stenosis < 50% in the contralateral artery) were also compared. In an attempt to determine risk factors associated with the ocular ischemic syndrome (OIS), the authors compared patients with severe OCAD and OIS with patients with severe OCAD without OIS. RESULTS: Peak systolic and end diastolic velocities in the ophthalmic, central retinal, and temporal short posterior ciliary arteries were significantly lower in patients with severe OCAD (P < 0.01). The mean resistive indices in the central retinal and temporal short posterior ciliary arteries were higher in the group with severe OCAD (P < 0.01). Similar results were obtained in the analysis of 25 patients with asymmetric carotid stenosis. Younger age (P = 0.012), severe bilateral OCAD (P = 0.01), high grade carotid stenosis (P = 0.013), and reversed ophthalmic artery flow (P = 0.038) were significant risk factors for OIS. CONCLUSIONS: Patients with severe OCAD show hemodynamic changes that suggest reduced retrobulbar blood flow. Patients with severe bilateral OCAD, high-grade carotid stenosis, and reversed ophthalmic artery flow may have a greater risk of developing OIS. PMID- 9400758 TI - A comparison of retrobulbar versus sub-Tenon's corticosteroid therapy for cystoid macular edema refractory to topical medications. AB - OBJECTIVE: The objective is to compare the effectiveness of retrobulbar and posterior sub-Tenon's injection of corticosteroids for treatment of post-cataract cystoid macular edema that was refractory to topical medications. DESIGN: A retrospective study was performed. PARTICIPANTS: A total of 48 patients (49 eyes) with post-cataract cystoid macular edema refractory to topical medications was studied. INTERVENTION: Patients received either a single retrobulbar injection (18 eyes) or 3 biweekly posterior sub-Tenon's injections (31 eyes) of corticosteroids. MAIN OUTCOME MEASURES: Patients were observed for clinical resolution of the cystoid macular edema, visual acuity, and intraocular pressure. RESULTS: Both treatment methods resulted in significant improvement in visual acuity. The posterior sub-Tenon's group had a visual improvement from 20/92 pretreatment to 20/50 post-treatment (P = 0.0001) with a median follow-up of 12 months. The retrobulbar group had a visual improvement from 20/97 pretreatment to 20/58 post-treatment (P = 0.035) with a median follow-up of 10 months. The visual improvement was not significantly different between the two groups. The average intraocular pressure increased from a pretreatment level of 14.1 mmHg to a high of 17.7 mmHg (P < 0.00005) in the sub-Tenon's group. The average intraocular pressure increased from 15.1 mmHg to a high of 17.6 mmHg (P = 0.04) in the retrobulbar group. CONCLUSIONS: Cystoid macular edema that persists after treatment with topical medications may improve after retrobulbar or posterior sub Tenon's corticosteroid injections. There was no significant difference in outcome between the two treatment groups. PMID- 9400759 TI - Outpatient postoperative fluid-gas exchange after early failed vitrectomy surgery for macular hole. AB - BACKGROUND: Vitrectomy surgery with fluid-gas exchange and prone positioning has been shown to close macular holes and improve vision. In those eyes that have failed surgery, repeat vitrectomy has been advocated. As an alternative, the authors performed an outpatient postoperative fluid-gas exchange on eyes when the macular hole failed to close after vitrectomy surgery. METHODS: The authors reviewed all cases of failed vitrectomy surgery for macular holes that underwent a postoperative fluid-gas exchange. Eyes were considered to have failed initial surgery if a rim of subretinal fluid surrounded an open full-thickness macular hole. RESULTS: Twenty-three consecutive eyes underwent outpatient fluid-gas exchange 1 week to 8 weeks after vitrectomy surgery. In 17 eyes (74%), fluid-gas exchange resulted in flattening and closure of the macular hole. In all 17 eyes, visual acuity improved 2 or more lines, with 8 (35%) of the 23 eyes achieving 20/50 or better visual acuity. CONCLUSIONS: Postoperative fluid-gas exchange may achieve successful closure of macular holes and improve vision in eyes that have failed surgery for full-thickness macular holes and should be considered as a cost-effective alternative to repeat vitrectomy. PMID- 9400760 TI - A multivariate analysis of anatomic success of recurrent retinal detachment treated with pneumatic retinopexy. AB - PURPOSE: The purpose of the study is to determine the success rate of pneumatic retinopexy (PR) after failed scleral buckling and to elucidate the predictors for anatomic failure by multiple logistic regression analysis. METHODS: Thirty-six eyes with recurrent retinal detachment after failed scleral buckling underwent PR. Intraocular tamponade was attained with SF6 (20 eyes), C3F8 (13 eyes), and air (3 eyes). Median follow-up was 14 months. RESULTS: Retinal reattachment was obtained in 69.4%. Multivariate analysis identified two risk factors for failure: location of retinal break either on the posterior slope or posterior to buckle (P = 0.01) and extent of retinal detachment greater than two quadrants (P = 0.02). CONCLUSIONS: Pneumatic retinopexy is an effective alternative to revision surgical operations if the leaking retinal break is located on the buckle. PMID- 9400761 TI - Quantitative analysis of macular holes with scanning laser tomography. AB - OBJECTIVE: The purpose of the study is to establish objective, quantitative, and reproducible three-dimensional analysis for macular holes with scanning laser tomography and to correlate measurements with visual acuity. DESIGN: The authors performed a cross-sectional, nonrandomized study. PARTICIPANTS: The authors examined 28 full-thickness macular holes of 23 patients aged 61 to 84 years. INTERVENTION: Confocal infrared imaging with scanning laser tomography using the TopSS (790 nm, 10 degrees field) three-dimensional measurements and macular hole analysis were performed. MAIN OUTCOME MEASURES: Area, depth, and volume parameters for both macular holes and rims were obtained in two ways: (1) reference plane for analysis equal to height of the retina (offset surface distance [OSD] 0) or (2) equal to height of the surrounding edematous rim of the hole (OSD user). Correlation of measurements with visual acuity and groups of macular holes was performed. Reproducibility from three image series per subject and intraobserver variability from ten measurements in four subjects were computed. RESULTS: Scanning laser tomography could detect macular hole and rim features in all subjects. Infrared images provided clinically useful information that may help explain macular hole pathophysiology. Based on quantitative, three dimensional measurements, holes were assigned to four groups: large, small, shallow, and average. Groups varied significantly (P < 0.05) for the majority of measurements. Visual acuity correlated significantly (P < 0.05) with macular hole volume, depth, slope, and rim height with OSD user only, but not with hole area. Holes computed with OSD user were deeper and of greater volume. Reproducibility was excellent for hole area, slope, and rim area; good for hole volume and depth; variable for rim volume; and improved with OSD user. Intraobserver variability was low in each group. CONCLUSIONS: Scanning laser tomography is a reproducible three-dimensional imaging technique providing objective and quantitative clinical information in assessing, grouping, and managing macular holes. By setting the OSD to rim height, additional information of rim height and hole volume was provided and correlated with visual acuity. In addition, more reliable differences among macular hole groups were found. Axial measurements such as macular hole depth, volume, and rim height may be more important for visual acuity than hole area indicating their possible predictive value for outcome measures. PMID- 9400762 TI - The treatment of macular disease using a micropulsed and continuous wave 810-nm diode laser. AB - OBJECTIVE: The purpose of the study is to determine whether the 810-nm diode wavelength using a rectangular waveform is clinically effective in the treatment of choroidal neovascularization from age-related macular degeneration and to determine whether macular edema secondary to branch vein occlusion or diabetic retinopathy can be effectively treated with this laser using the micropulse waveform. DESIGN: Review of consecutive nonrandomized patients whose eyes were treated with the diode laser over a 30-month period. PARTICIPANTS: Fifty-three patients with an initial presentation of choroidal neovascularization located subfoveally (77%), extrafoveally (17%), and juxtafoveally (6%); 14 patients with macular edema from a branch vein occlusion; and 59 patients with diabetic macular edema, 40 of which were treated for the first time. INTERVENTION: Ablative rectangular wave laser photocoagulation was applied to the choroidal neovascular membranes and very light threshold treatment was applied in a macular grid to treat retinal edema. Microaneurysms were not targeted. MAIN OUTCOME MEASURES: Anatomic resolution of macular edema or choroidal neovascularization and visual acuity. RESULTS: Sixty percent of eyes treated for choroidal neovascularization had no persistence or recurrence at 6 months, and 72% achieved visual stabilization. In 8% of eyes, some localized bleeding occurred during photocoagulation. Clinical resolution of macular edema from branch vein occlusion occurred by 6 months in 92% of eyes, and 77% had stabilization of visual acuity. At 6 months, 76% of newly treated patients with diabetic macular edema and 67% of previously treated patients had clinical resolution of their edema. Vision was improved or stabilized in 91% and 73% of newly treated and retreated patients at 6 months, respectively. CONCLUSIONS: The micropulsed 810-nm diode laser is clinically effective in the treatment of macular edema from venous occlusion and diabetic retinopathy, and the rectangular (normal) mode diode laser can be used in many eyes with choroidal neovascularization. PMID- 9400763 TI - Visual acuity outcomes among patients with appositional suprachoroidal hemorrhage. AB - OBJECTIVE: The purpose of the study is to investigate visual acuity outcomes among patients with appositional suprachoroidal hemorrhage and to identify clinical features associated with visual prognosis. DESIGN: The study design was a retrospective chart review. PARTICIPANTS: All patients whose ocular echographic examination results showed appositional suprachoroidal hemorrhage at the Bascom Palmer Eye Institute between January 1, 1987, and December 31, 1996 were included. Fifty-one patients were identified. INTERVENTION: Demographic and clinical data were abstracted from patients' medical records. MAIN OUTCOME MEASURES: Visual acuity at 3, 6, and 12 months posthemorrhage and clinical features associated with visual prognosis were defined. RESULTS: At final follow up fifteen (29.4%) patients achieved either their prehemorrhage visual acuity (n = 7) or a visual acuity of 20/200 or better (n = 8), but 14 (27.5%) patients had no light perception. Predictors of a poor visual outcome include vitreous incarceration in the wound/bleb (P = 0.014), concurrent or delayed retinal detachment (P = 0.003), and afferent pupillary defect on presentation (P = 0.002). Poorer visual acuity on presentation (r = 0.37, P = 0.008) and longer duration of central retinal apposition (r = 0.51, P < 0.001) also were significantly associated with poor final visual acuity. Patients in whom the suprachoroidal hemorrhage maintained an appositional configuration for more than 14 days were more likely to have worse final visual acuities than were patients with appositional choroidals for fewer than 14 days (P = 0.006). The association between duration of apposition and final visual acuity was significant, both among patients whose suprachoroidal hemorrhages were observed (n = 26, r = 0.60, P = 0.001) and among patients who underwent secondary surgical intervention (n = 23, r = 0.66, P = 0.001). Patients with postoperative suprachoroidal hemorrhages achieved better final visual acuities than did patients in whom suprachoroidal hemorrhages developed intraoperatively or after trauma (P = 0.038). CONCLUSIONS: Appositional suprachoroidal hemorrhage is a serious ocular complication with a guarded visual prognosis. A variety of clinical features, including vitreous incarceration in the wound/bleb, concurrent or delayed retinal detachment, afferent pupillary defect, presenting visual acuity, and duration of central retinal apposition, may help predict visual outcome. PMID- 9400764 TI - Vitreous surgery for chronic macular holes. AB - OBJECTIVE: The purpose of the study is to compare the results of vitreous surgery for a group of patients with chronic macular holes with a group of patients with acute-onset macular holes undergoing identical surgery. DESIGN: A case-control study design was used. PARTICIPANTS: The duration of symptoms of visual loss due to macular holes was greater than 1 year's duration in 11 eyes in each group consecutively operated on within a few days. INTERVENTION: All patients underwent macular hole surgery. MAIN OUTCOME MEASURES: Ophthalmoscopic resolution of the macular hole, improvement of 2 lines of visual acuity or greater, improvement in mean and median visual acuity, and rate of 20/40 or greater final visual acuity. RESULTS: The hole resolved in 9 of 11 eyes in the chronic group and 10 of 11 eyes in the acute group. The mean (median) preoperative visual acuity was 20/151 (20/200) in the chronic group and 20/139 (20/200) in the acute group. The 3-month mean (median) postoperative visual acuity was 20/85 (20/80) in the chronic group and 20/62 (20/63) in the acute group. The final mean (median) postoperative visual acuity was 20/96 (20/ 100) in the chronic group and 20/48 (20/50) in the acute group (P = 0.022). The mean interval to final follow-up examination was 70 weeks for the chronic group and 44 weeks for the acute group. Five (45%) of 11 eyes with chronic holes and 8 (73%) of 11 eyes in the acute group had a final visual acuity of 2 lines or better than the preoperative visual acuity. Cataract extraction had been performed by the final follow-up examination in 7 chronic eyes (64%) and 2 acute eyes (18%). CONCLUSIONS: Chronic macular holes have a similar anatomic success rate, but a poorer visual prognosis than acute holes after macular hole surgery. Vitreous surgery benefits some patients with idiopathic macular holes of greater than 1 year's duration. PMID- 9400765 TI - Laser photocoagulation for retinal detachments and retinal tears in cytomegalovirus retinitis. AB - OBJECTIVE: Retinal detachment complicates the course of cytomegalovirus (CMV) retinitis in nearly 30% of human immunodeficiency virus-infected patients. The study goal was to evaluate laser photocoagulation in the treatment of CMV retinitis-related retinal detachments and retinal tears. DESIGN: Nonrandomized, observational cohort study. PARTICIPANTS: Sixty-three patients with CMV retinitis related retinal detachments and nine patients with peripheral retinal tears in eyes with CMV retinitis were studied. INTERVENTION: Of the 63 eyes with retinal detachment, 23 patients were treated with demarcating laser photocoagulation, 24 patients underwent vitrectomy with silicone oil, and 16 patients were observed without treatment. All nine patients with peripheral retinal tears received laser photocoagulation. MAIN OUTCOME MEASURES: Time to progression of retinal detachment, final visual acuity, and need for vitrectomy surgery were studied. RESULTS: Median time to progression of the retinal detachment in the laser treated patients was 175 days versus 39 days in observed patients (P = 0.012). Both initial (P < 0.001) and final (P = 0.005) visual acuities were better in the patients with laser-treated detachment than in the observed or vitrectomy patients. The retinal detachment groups were comparable in follow-up, zone and location of detachment, and size of holes, but the vitrectomy and observed groups had more cases with extensive CMV retinitis. Vitrectomy surgery was required in 9 of 16 (56%) in the observed group and 7 of 23 (30%) in the laser group. Two of nine patients (22%) who failed to respond to laser treatment for retinal breaks required vitrectomy surgery. CONCLUSIONS: Laser photocoagulation of selected retinal detachments and retinal tears delayed or avoided vitrectomy with silicone oil. It may be an important treatment modality for patients with nonmacular detachments and for those who are receiving local anti-CMV therapy with intravitreal injections or pellets, in whom silicone oil may affect the efficacy of the local treatment. PMID- 9400766 TI - Laser photocoagulation repair of macula-sparing cytomegalovirus-related retinal detachment. AB - OBJECTIVE: The purpose of the study is to investigate the role of laser photocoagulation in the treatment of macula-sparing cytomegalovirus (CMV)-related retinal detachment (CMVRD) in patients with acquired immune deficiency syndrome (AIDS). DESIGN: Seven macula-sparing CMVRD identified between July 1995 and February 1997 were managed with laser photocoagulation and observed prospectively (group I). Seven CMVRD reattached with pars plana vitrectomy (PPV) and silicone oil injection (group II) between January 1992 and June 1996 were analyzed retrospectively. PARTICIPANTS: Patients with AIDS with macula-sparing rhegmatogenous CMVRD with no proliferative vitreoretinopathy and visual acuity better than 20/30 were studied. INTERVENTION: Demarcation laser photocoagulation (group I) or PPV with silicone oil injection (group II) was performed. MAIN OUTCOME MEASURES: Postoperative best-corrected visual acuity (BCVA), temporary or permanent visual loss, CMVRD progression or recurrence, and cataract were measured. RESULTS: Follow-up ranged from 2 to 19 months (mean, 9 months) in group I. Post-treatment BCVA was unchanged in all eyes after laser. One retina redetached 9 months after laser treatment. Final visual acuity was less than 20/40 in one eye because of progressive CMV retinitis. Follow-up ranged from 2 to 24 months (mean, 10.4 months) in group II. All group II RDs were reattached successfully with PPV and silicone oil injection. Best-corrected visual acuity was an average of 1.6 lines worse after vitrectomy. Silicone-induced hyperopic shift caused temporary visual loss in all eyes (mean duration, 5.6 weeks). Delayed visual loss due to cataract formation occurred in five eyes. Three eyes had cataract extraction within 6 months. Two partial redetachments developed. One was repaired with repeat vitrectomy. Final visual acuity was less than 20/40 in five of seven eyes because of progressive CMV retinitis (1), dense cataract (2), uncorrected refractive error (2), and uncertain cause (1). CONCLUSIONS: Demarcation laser photocoagulation appears to be an effective treatment for many macula-sparing CMVRD. Loss of BCVA, temporary postoperative visual loss due to silicone-induced refractive error, and delayed visual loss due to cataract after vitrectomy with silicone oil injection may be avoided. Demarcation laser photocoagulation may be an effective alternative to vitrectomy in macula-sparing CMVRD. PMID- 9400768 TI - Analysis of videokeratography after penetrating keratoplasty: topographic characteristics and effects of removing running sutures. AB - OBJECTIVE: Previous studies have shown that removal of running sutures after penetrating keratoplasty causes unpredictable changes in astigmatism. The current study was conducted to investigate whether computer-assisted videokeratography is beneficial for predicting visual outcomes after running sutures are removed. DESIGN: The design was that of a prospective clinical study. PARTICIPANTS: The authors prospectively studied 29 consecutive eyes undergoing a 10-0 nylon running suture removal after penetrating keratoplasty. INTERVENTIONS: Videokeratography was performed before, 1 week, 1 month, and 3 months after removal of sutures. MAIN OUTCOME MEASURES: Changes in refractive and topographic astigmatism after suture removal were measured. Topographic patterns and their quantitative descriptors also were analyzed. RESULTS: An asymmetric bowtie was the most common videokeratography pattern both before and after suture removal. After suture removal, the incidence of peripheral corneal steepening increased significantly (2 vs. 21 eyes, P < 0.0001), and that of focal flattening of the midperipheral cornea decreased (13 vs. 5 eyes, P = 0.046). The mean topographic astigmatism, surface regularity index, and corrected visual acuity were improved significantly by suture removal in eyes that had localized flattening but not in eyes without this finding. Eyes having either skewed axis in astigmatism or topographic astigmatism of more than 9 diopters also showed significant decreases in astigmatism. CONCLUSIONS: Suture removal after keratoplasty is advantageous for both reducing astigmatism and normalizing topography, especially in eyes that have localized flattening of the midperipheral cornea. Predictability of visual outcomes of a running suture removal in postkeratoplasty eyes may be improved by the use of videokeratography. PMID- 9400767 TI - Amniotic membrane transplantation for ocular surface reconstruction in patients with chemical and thermal burns. AB - OBJECTIVE: The purpose of the study is to examine the usefulness of preserved human amniotic membrane transplantation in patients with chemical and thermal burns. DESIGN: The study design was a nonrandomized clinical trial. PARTICIPANTS: Seven eyes of six patients with severe chemical (n = 5) and thermal (n = 2) burns were studied. INTERVENTION: Eyes were treated with excision of cicatricial tissues followed by a placement of amniotic membrane on the sclera. Transplantation of limbal grafts from an opposite eye (n = 4) or from donor eyes preserved at -80 degrees C (n = 2) was performed simultaneously. MAIN OUTCOME MEASURES: Reconstruction of ocular surface epithelia and visual acuity were measured. RESULTS: With the mean observation period of 53.3 weeks, central corneal epithelium was reconstructed successfully in all eyes. Neither amniotic membrane nor limbal grafts were rejected. A persistent epithelial defect developed in one eye, which was treated successfully by tarsorrhaphy. After surgery, the corneal epithelium showed normal arrangements on specular microscopy, and its barrier function recovered to seminormal. Corrected visual acuity markedly improved in each eye. Regenerated conjunctiva on the amniotic membrane was stable and uninflammed with minimum-to-mild scarring. Slight recurrence of conjunctivalization was noted in three eyes. However, because these eyes were stable and central cornea was clear, no further surgery was needed. CONCLUSIONS: Amniotic membrane transplantation promotes normal conjunctival epithelialization while suppressing fibrosis formation. The procedure, especially when performed with limbal autograft transplantation, appears to be effective for the treatment of chemical or thermal burns of the ocular surface. PMID- 9400769 TI - Mitomycin C treatment for conjunctival-corneal intraepithelial neoplasia: a multicenter experience. AB - OBJECTIVE: The purpose of the study is to evaluate the efficacy and risks of topical mitomycin C (MMC) for conjunctival-corneal intraepithelial neoplasia (CCIN). DESIGN: The study design was a clinical case series of CCIN. PARTICIPANTS: Seventeen patients, 16 with biopsy-confirmed CCIN and 1 with invasive squamous cell carcinoma (SCC), were included in the study. INTERVENTION: Patients received topical drops of MMC 0.02% to 0.04% four times daily from 7 to 28 days. Retreatment was done in cases of lesion recurrence. MAIN OUTCOME MEASURES: The size of the CCIN before and after the treatment and ocular complications post-MMC application were evaluated. RESULTS: Ten patients remained disease-free after one course of MMC application. In one case, residual CCIN remained very small without regrowth. In the one patient with invasive SCC and in five patients with CCIN, regrowth occurred within 6 months of the first treatment. After retreatment, invasive SCC and CCIN in an additional two patients were eradicated. In two cases, although the size of the lesions decreased after two and three applications of MMC, regrowth occurred, and the CCIN returned to its original size. In the final case, limited recurrence has occurred and no retreatment has been done. The complications of MMC use included mild-to-moderate conjunctival hyperemia and mild allergy, which resolved after discontinuation of the treatment. Severe pain manifested when treatment was longer than 14 days. CONCLUSIONS: Application of topical MMC is an efficient treatment for most but not all cases of CCIN. PMID- 9400770 TI - Orbital involvement in allergic fungal sinusitis. AB - BACKGROUND: Although allergic fungal sinusitis is a relatively common, noninvasive form of paranasal sinus mycosis, and despite frequent orbital involvement, there have been few reports of this condition in the ophthalmic literature. METHODS: Two cases of allergic fungal sinusitis having orbital symptoms are described. The current classification, typical presentation, and ideal management of fungal sinusitis are reviewed. RESULTS: Distinguishing radiologic and pathologic features were present in both patients. Aspergillus flavus was cultured in one case, and Bipolaris spicifera was cultured in the other. CONCLUSIONS: Allergic fungal sinusitis is a unique subset of sino-orbital disease with highly characteristic clinical, radiologic, and pathologic features. Unlike invasive forms of mycotic disease, allergic fungal sinusitis may be managed adequately with surgical debridement, aeration of the involved sinuses, and systemic and topical corticosteroids. PMID- 9400771 TI - Combined chemoreduction and adjuvant treatment for intraocular retinoblastoma. AB - OBJECTIVE: The purpose of the study is to investigate chemoreduction and adjuvant treatment (AT) for retinoblastoma and its effect on complete retinal tumor control, vitreous seed control, and subretinal seed control. DESIGN: The study design was a prospective, nonrandomized clinical trial. PARTICIPANTS: There were 130 intraocular retinoblastomas in 52 eyes of 32 consecutive patients observed for at least 1 year after initiation of treatment. INTERVENTION: Treatment with chemoreduction using vincristine, etoposide, and carboplatin (VEC) and adjuvant treatment (+ AT) (cryotherapy, laser photocoagulation, thermotherapy, chemothermotherapy, plaque radiation therapy, or external beam radiation therapy) were assessed. MAIN OUTCOME MEASURES: The effect of chemoreduction for 6 cycles (VEC x 6) versus fewer than 6 cycles (VEC x <6) on retinoblastoma control was analyzed. Furthermore, the impact of adjuvant treatment (+ AT) versus no adjuvant treatment (no AT) on retinoblastoma control was analyzed. RESULTS: Retinal tumors showed favorable initial regression with chemoreduction. Adjuvant treatment was applied to 93% of the retinal tumors after chemoreduction and only 2% recurred over the mean follow-up of 17 months (range 13-27 months). Vitreous seeds and subretinal seeds showed initial regression and often complete disappearance with chemoreduction. In those eyes with seeds before treatment, the addition of AT to VEC for 6 cycles decreased the vitreous seed recurrence from 75% to 0% (P = 0.04) and also decreased the subretinal seed recurrence from 67% to 0% (P = 0.003). More important, when considering that enucleation or external beam radiation therapy was the only other treatment option for these 52 eyes, the authors were successful in avoiding these methods in 42% of cases. Of the 36 eyes classified as Reese-Ellsworth group 5, there was 78% ocular salvage, and external beam radiation therapy was avoided in 25% of these eyes. There was a 100% ocular salvage in the group 5 eyes that received VEC for 6 cycles + AT to retinal tumors and seeds. CONCLUSIONS: Chemoreduction and AT to intraocular retinoblastoma and its seeds provides good retinal tumor control, even in eyes with advanced disease. Chemoreduction alone generally is not adequate to achieve complete tumor seed control. Cautious follow-up of affected patients is recommended because the risk for recurrent vitreous and subretinal seeds is substantial and proper treatment is critical for salvaging the eye. PMID- 9400772 TI - Ocular-hypertensive response to topical steroids in children. AB - OBJECTIVE: The purpose of the study is to investigate the rate and degree of ocular-hypertensive response to topical steroids in Chinese children. DESIGN: The study design was an institutional, randomized, clinical trial. PARTICIPANTS: A total of 19 consecutive patients were studied. INTERVENTION: Topical steroids were administered to Chinese children younger than 10 years of age who underwent bilateral strabismus surgery. One eye was randomized to receive topical 0.1% dexamethasone (DMS), whereas the fellow eye received 0.1% fluorometholone (FML) six times per day for up to 4 weeks. Intraocular pressure (IOP) was measured on the day before operation and at postoperative days 1, 3, 6, 10, 13, and 27, then every 2 weeks thereafter until the IOP fell to preoperative levels. Topical steroids would be stopped if IOP was 30.00 mmHg or greater. MAIN OUTCOME MEASURES: Peak IOP and maximal change of IOP from baseline were measured and categorized into low, intermediate, and high levels. Time to peak IOP also was studied. RESULTS: A total of 16 patients were included. The peak IOP for DMS treated eyes was 30.66 +/- 8.35 mmHg (range, 13.00-48.00 mmHg), whereas that in FML-treated eyes was significantly lower at 20.66 +/- 6.03 mmHg (range, 11.30 36.30 mmHg) (P = 0.001). The maximal change in IOP ranged from -2.60 to +31.00 mmHg in DMS-treated eyes (mean, 15.48 +/- 8.71 mmHg), almost double that of FML treated eyes (range, +1.00 to +17.00 mmHg; mean, 5.83 +/- 4.96 mmHg) (P = 0.001). When the ocular-hypertensive responses of both DMS and FML groups were categorized into three levels of severity, significant differences were found between the two treatment groups (P = 0.001). In the DMS group, nine patients (56.25%) were high responders and six patients (37.5%) were intermediate responders. In the FML group, only one patient (6.25%) was a high responder. CONCLUSIONS: The ocular-hypertensive response to topical DMS in children occurs more frequently, more severely, and more rapidly than that reported in adults. A total of 56% of the studied children, all younger than 10 years of age, were high responders to topical DMS. Of these, 89% attained their peak IOP within 8 days. Its use in children should best be avoided if possible. It would be desirable to monitor the IOP when it is being used. Conversely, FML produced a much less ocular-hypertensive effect and therefore poses an acceptable risk of clinically significant pressure elevation. PMID- 9400773 TI - Bleb-related ocular infection in children after trabeculectomy with mitomycin C. AB - OBJECTIVE: The purpose of the study is to report the clinical course of bleb related ocular infection in children after trabeculectomy with adjunctive mitomycin C. DESIGN: The study design was a retrospective review of all patients with a diagnosis of bleb-related ocular infection after trabeculectomy with adjunctive mitomycin C. PARTICIPANTS: Three children were identified in whom late postoperative bleb-related ocular infection developed. INTERVENTION: Treatment consisted of vitreous biopsy with intravitreous antibiotic and corticosteroid injection and/or bleb culture with topical and intravenous antibiotic administration. MAIN OUTCOME MEASURES: Visual acuity and intraocular pressure were measured. RESULTS: Bleb-related ocular infection developed an average of 16.7 +/- 10.9 months after trabeculectomy (range, 4-23 months). The mean age at presentation was 7.0 +/- 2.6 years (range, 4-10 years). Vitreous cultures were positive for staphylococci in two cases. A bleb culture from the third case also grew staphylococcus. All of the children recovered their initial vision after treatment of infection. However, one lost six lines of vision after a subsequent retinal detachment. Additional glaucoma surgery was required in one patient. CONCLUSIONS: Late bleb-related ocular infection may occur in children after trabeculectomy with mitomycin C and is characterized by abrupt onset, bleb infiltration, and rapid progression. Despite early preservation of vision after treatment of infection, significant late visual loss can occur. PMID- 9400774 TI - Clinical experience of trabeculotomy for the surgical treatment of aniridic glaucoma. AB - OBJECTIVE: The purpose of this study is to determine the efficacy of initial trabeculotomy in the patient with aniridic glaucoma. DESIGN: Clinical charts were reviewed. PARTICIPANTS: Twenty-nine eyes of 16 patients with aniridia were studied. INTERVENTION: Glaucoma surgery was performed. As an initial procedure, trabeculotomy was performed in 12 eyes, other surgery was performed in 17 eyes (trabeculectomy, 5; goniotomy, 5; other, 7). MAIN OUTCOME MEASURES: Success was defined as an intraocular pressure (IOP) of 21 mmHg or lower, and no further surgery was performed. RESULTS: Ten (83%) of 12 eyes obtained IOP control after first (6 eyes) or second (4 eyes) trabeculotomy with a mean follow-up period of 9.5 years. Five eyes maintained visual acuity of 20/40 to 20/200. No serious complications were found after trabeculotomy. Three (18%) of 17 eyes were controlled with the first glaucoma surgery other than trabeculotomy (goniotomy, trabeculectomy, trabeculectomy combined with trabeculotomy, and Molteno implant). Good IOP control was obtained in 8 (47%) of 17 eyes after several surgeries with a mean follow-up period of 10.4 years. Four of 17 eyes became phthisical. CONCLUSION: This study suggests that trabeculotomy is the preferred initial operation for uncontrolled glaucoma with aniridia. PMID- 9400775 TI - The effect of adjunctive mitomycin C in Molteno implant surgery. AB - PURPOSE: The purpose of the study is to assess the effect of adjunctive intraoperative mitomycin C (MMC) in Molteno drainage device implantation for patients with recalcitrant glaucomas. METHOD: Forty-nine eyes of 49 patients who underwent one-stage, single-plate Molteno device implantation with adjunctive intraoperative MMC (0.5 mg/ml) for 3 to 5 minutes (MMC group) were compared to a historic control group of 51 eyes of 51 patients (control group) who received one stage, single-plate Molteno device implantation without MMC. Success (survival) was defined as an intraocular pressure (IOP) between 6 and 21 mmHg, inclusive, with (qualified success) or without (complete success) glaucoma medications and with no additional glaucoma surgery, phthisis, implant removal, or loss of light perception. RESULTS: Preoperative conditions were similar between the two groups. There was no significant difference in surgical survival rate between the two groups (P = 0.13, log-rank test). There also were no significant differences in the postoperative IOP levels and numbers of antiglaucoma medications between the two groups at all times (P > 0.05). Visual acuity was improved or remained within one line of preoperative visual acuity in 76.1% of the MMC group and 78.7% of the control group at 1 year after surgery (P = 0.76, chi-square test). Complications and reoperation for complications were similar in both groups (P > 0.05, chi square test) except for the incidence of early postoperative hypotony and the total number of eyes with complications not requiring reoperation, which were more common in the MMC group (P = 0.027, 0.005, respectively, chi-square test). The most common complications included hypotony with or without a flat anterior chamber or choroidal detachment, followed by hyphema and tube plugging. CONCLUSION: Molteno device implantation with adjunctive intraoperative MMC in patients with complicated glaucoma may not offer a better chance of surgical success compared with Molteno implantation without MMC. PMID- 9400776 TI - Pattern electroretinogram and spatial contrast sensitivity in primary congenital glaucoma. AB - OBJECTIVE: The authors investigated the temporal and spatial characteristics of pattern electroretinogram (PERG) and spatial contrast sensitivity (CS) in primary congenital glaucoma (PCG) to determine whether the PERG and CS could be useful tools in the diagnosis of childhood glaucoma, especially PCG. PARTICIPANTS: The PERGs were evaluated in eyes from ten patients with PCG and nine age-matched visually normal subjects. INTERVENTION: All patients received complete ophthalmologic evaluations including visual field testing. MAIN OUTCOME MEASURES: The PERGs were recorded using phase-alternating (2, 4, and 16 reversals per second [rps]) checkerboard patterns (30' and 60' checks). RESULTS: The patients with PCG exhibited decreased CS when compared with that of control subjects. Significant PERG deficits also were detected in these patients. However, PERG amplitude in patients with PCG almost reached control subject levels at high (16 rps) temporal frequency. This was true for both 30' and 60' checks. Taken together, these observations on PERG amplitude suggest a more important deficiency of the neural response of the retinal cells at lower temporal frequency (rps) in patients with PCG. This is unlike primary open-angle glaucoma (POAG) in which significant PERG deficits are observed at high temporal frequencies. CONCLUSIONS: The PERG amplitude is reduced in patients with PCG, and this is consistent with a loss of CS and visual field changes in these patients. However, the spatiotemporal characteristics of the PERG deficits in PCG differ from those of POAG. This could suggest a difference in the mechanisms mediating retinal ganglion cell dysfunction in the two types of glaucoma. PMID- 9400777 TI - Full-time atropine, intermittent atropine, and optical penalization and binocular outcome in treatment of strabismic amblyopia. AB - OBJECTIVE: The purpose of the study is to evaluate the monocular and binocular outcome of three types of "penalization" (blurring of the sound eye) treatment of amblyopia: traditional full-time atropine or optical penalization and a new intermittent atropine regimen involving atropine instillation 1 to 3 days a week. DESIGN: The study design was a retrospective study. PARTICIPANTS: A total of 163 patients with strabismic amblyopia treated by full-time atropine (n = 38), intermittent atropine (n = 73), or optical (n = 52) penalization participated. MAIN OUTCOME MEASURES: Logarithm of the minimum angle of resolution (logMAR) visual acuity, and binocularity index were determined. RESULTS: All three forms of penalization produced statistically significant mean reduction in amblyopia (1.7-2.7 logMAR lines) and mean improvement in binocularity by the end-of treatment or long-term follow-up visit or both, with minimal mean loss after discontinuation or slight mean improvement on these measures at long-term mean follow-up of 1.9 to 4 years across groups. Few patients achieved high-grade stereoacuity. Compliance was high. Comparable efficacy was found for all three treatment groups after controlling for age, depth of amblyopia, and binocularity at the initial visit. Initial-visit amblyopia depth was strongly and significantly associated with amblyopia depth at both post-treatment visits. Pretreatment and post-treatment binocularity showed a similar strong relationship. Surprisingly, however, there was no consistent or significant association found between depth of amblyopia and binocularity in any visit combination. Post-treatment measures of these two variables also were not associated with initial-visit age or refractive error at any clinically significant level. Mean treatment duration was 1.1 to 2.9 years and was not found to be associated with visual outcome. Amblyopia reversal was found in one (full time atropine) case at a clinically important level. CONCLUSIONS: The authors confirmed previous reports of penalization's efficacy as a primary treatment of moderate amblyopia (20/100 or better acuity) and, in some cases, relatively severe amblyopia (>20/100) and also confirmed its ability to significantly improve mean binocularity. Amblyopia and binocularity appear to respond to treatment independently and, within the postinfancy age range of the sample studied, the responses appear to be independent of initial-visit age. The high acceptability to patients and parents of atropine penalization, and particularly of the intermittent regimen introduced here, suggests the need for prospective study-based re-evaluation of the relative merits of penalization and occlusion as the standard of care for mild-to-moderate amblyopia. PMID- 9400778 TI - Penalization versus part-time occlusion and binocular outcome in treatment of strabismic amblyopia. AB - OBJECTIVE: The purpose of the study is to compare the visual outcome of occlusion versus penalization treatment of strabismic amblyopia, with particular attention to binocularity outcome. DESIGN: The study design was a retrospective study. PARTICIPANTS: Patients with strabismic amblyopia, 75 receiving penalization alone, 87 with a history of occlusion treatment who were later treated by penalization, and 30 treated by means of part-time occlusion (2 to 6 hours/day) participated in this study. MAIN OUTCOME MEASURES: Logarithm of the minimum angle of resolution (logMAR) visual acuity and binocularity index were measured. RESULTS: No statistically significant difference was found between outcomes for the penalization groups with and without a history of occlusion, either by univariate analysis or by multivariate analysis controlling for initial-visit age, acuity, and binocularity status. One marginally significant outcome difference was found between the pure penalization and part-time occlusion groups by univariate analysis, but no significant difference was found in the multivariate analyses controlling for the same three variables at the initial visit. All visual outcome differences between the pure penalization and part-time occlusion groups were less than 1 logMAR line visual acuity or less than a half unit on the binocularity index. CONCLUSIONS: The study provided no evidence of a difference in visual function outcome between penalization and occlusion, in terms of either statistical or clinical significance, although limitations of the patient samples used preclude these data from showing conclusively that there was no such difference. The lack of any other study adequately comparing these two treatment methods, in combination with the current study's demonstration of the difficulty of making adequate retrospective-based comparison despite a large patient base (n = 1413), suggests that a large prospective, randomized comparative treatment trial is needed. If atropine penalization, with its high acceptability to patients and parents, is found to produce results comparable with those of occlusion in cases of mild-to-moderate amblyopia, as the current and previous smaller studies suggest, then reconsideration of the standard of care for such amblyopia cases is indicated. PMID- 9400779 TI - Automated pupil perimetry in amblyopia: generalized depression in the involved eye. AB - OBJECTIVE: This study was designed to determine whether the relative afferent pupillary defects observed commonly in amblyopic eyes are associated with a uniform depression of the pupillary light reflex throughout the visual field or solely by a focal decrease in pupillary response near fixation. DESIGN: The authors used pupil perimetry to evaluate the contraction amplitude of the pupil in response to focal light stimuli at 76 points throughout the 30 degrees field in each eye of 28 patients with amblyopia. The "pupil fields" were recorded using a computerized infrared pupillograph linked to a Humphrey Field Analyzer, so that the pupil contraction could be recorded in response to perimetric light stimuli. PARTICIPANTS: Nine patients had strabismic amblyopia, ten had anisometropia, six had a combination of anisometropia and strabismus, and three had deprivation amblyopia due to monocular congenital cataract. MAIN OUTCOME MEASURES: Mean pupillary contraction amplitude for the entire field and focal amplitudes at each tested location were compared. Mixed-model analysis of variance was used to assess effects of perimetry location, type of amblyopia, and interaction effects. RESULTS: The overall average of all the pupil contractions throughout the 30 degrees field was less for the amblyopic eye compared with that of the fellow eye. This decrease in focal pupil response for amblyopic eyes was present in each type of amblyopia and was greatest for deprivation amblyopia. The contraction amplitude was depressed diffusely throughout the pupil field and showed neither focal deficits nor a selective depression about fixation. CONCLUSION: Amblyopia produces a global depression of focal pupillary responses across the entire 30 degrees field. PMID- 9400780 TI - Posterior capsulectomy in pediatric cataract surgery: the necessity of a choice. AB - OBJECTIVE: The purpose of the study is to evaluate whether a posterior capsulectomy combined with anterior vitrectomy is a necessity in pediatric cataract. DESIGN: The incidence of posterior capsule opacification, the need for additional surgical interventions, and the influence of a primary posterior capsulectomy after cataract surgery in children were evaluated. The analysis was carried out by studying patients' records retrospectively or after prospective follow-up. PARTICIPANTS: In 94 eyes (69 aphakic and 25 pseudophakic), the medical records were studied retrospectively. Twenty-eight eyes (18 aphakic and 10 pseudophakic) were observed prospectively during 1 year after surgery. In 20 eyes (6 aphakic and 14 pseudophakic) of 10 patients with bilateral cataract, a prospective comparison between the 2 eyes of the same patient also was carried out. INTERVENTION: Cataract surgery through the limbus with or without a primary posterior capsulectomy was performed in 114 eyes (43 of these received a posterior chamber intraocular lens [IOL] and 71 remained aphakic). In 28 eyes, the surgery was carried out by way of the pars plana (6 eyes received an anterior chamber IOL and 22 remained aphakic). MAIN OUTCOME MEASURES: Incidence of posterior capsule opacification, the need for secondary surgical intervention, and visual acuity were measured. RESULTS: Opacification of the posterior capsule is observed in all children's eyes when a primary posterior capsulectomy (combined with an anterior vitrectomy) was not carried out. Earlier secondary cataract formation is associated with a younger age and with implantation of an IOL. Eyes undergoing a primary opening of the posterior capsule during the initial surgery of children with bilateral cataract achieved, in most cases, a better visual acuity than did their fellow eyes. CONCLUSION: Although possibly a choice in older children, a primary posterior capsulectomy combined with anterior vitrectomy is a must in younger children and particularly when implantation of an IOL is planned. PMID- 9400781 TI - Trichotillomania. AB - PURPOSE: Trichotillomania is characterized by an irresistible urge to pull one's hair, and may involve the eyelashes or eyebrows. The authors present four cases of trichotillomania, and review the management of this unusual disorder. METHODS: The cases of four patients with trichotillomania were reviewed retrospectively. RESULTS: All four patients had characteristic areas of broken lashes along the lid in the absence of other signs of disease. Three of the four knew they were plucking the hair, yet could not control it. In the fourth, it was only after a lengthy observation period that she was discovered plucking. CONCLUSIONS: Trichotillomania has been infrequently reported in the ophthalmic literature. Management can be difficult. Many of these patients are aware of their behavior, but are unable to curtail it. Others may conceal or deny their habit. Psychiatric counseling may be of some benefit if patients are willing to undergo it. PMID- 9400782 TI - Parapapillary chorioretinal atrophy in patients with ocular hypertension. I. An evaluation as a predictive factor for the development of glaucomatous damage. AB - OBJECTIVE: To determine whether parapapillary chorioretinal atrophy is a risk factor for the development of glaucomatous optic disc or visual field damage. METHODS: The initial morphometric parameters of the optic disc and parapapillary atrophy were retrospectively investigated in 350 eyes of 175 patients with ocular hypertension. The prognostic value of parapapillary atrophy at the baseline examination and its relationship with known risk factors for the development of glaucomatous damage were analyzed by multivariate analysis. RESULTS: Visual field loss, optic disc damage, or both were detected in 98 eyes of 53 patients during the follow-up period of at least 10 years. By univariate analysis, the presence of parapapillary atrophy, as well as higher parapapillary atrophy area-disc area, zone beta area-disc area, and parapapillary atrophy length-disc circumference ratios, at the baseline examination was associated with the conversion to glaucoma. In addition, higher intraocular pressure, larger vertical cup-disc ratio, and smaller neural rim area-disc area ratio at the baseline examination were associated with subsequent glaucomatous optic nerve damage. In a multivariate regression model adjusted for other factors, intraocular pressure (relative risk, 1.19), neural rim area-disc area ratio (relative risk, 0.72), and zone beta area-disc area ratio (relative risk, 1.32) were found to be associated with the development of optic disc damage, visual field damage, or both. CONCLUSION: The presence and the size of parapapillary atrophy are related to the development of subsequent optic disc or visual field damage in patients with ocular hypertension. PMID- 9400783 TI - Parapapillary chorioretinal atrophy in patients with ocular hypertension. II. An evaluation of progressive changes. AB - OBJECTIVE: To determine whether parapapillary chorioretinal atrophy in patients with ocular hypertension remained stationary or progressed along with glaucomatous optic nerve damage. METHODS: The morphometric parameters and progression of parapapillary atrophy were retrospectively investigated, using serial photographs, in 350 eyes of 175 patients with ocular hypertension. The association of parapapillary atrophy progression with subsequent glaucomatous conversion and with other baseline patient- and eye-specific characteristics was analyzed. RESULTS: Progression in the area and extension of parapapillary atrophy before noticeable optic disc or visual field changes was observed in 48 (49.0%) of 98 eyes that converted to glaucoma, while parapapillary atrophy progression was noted in 25 (9.9%) of 252 ocular hypertensive eyes that did not develop glaucomatous damage (P<.001). The predictive sensitivity and specificity of this observation were 49% and 90%, respectively. In a logistic multiple regression model, the progression of parapapillary atrophy was associated with a family history of glaucoma (odds ratio, 2.7) and the initial size of zone beta (odds ratio, 1.64, for an increase of 0.10 of the zone beta area-disc area ratio). CONCLUSION: The progression of parapapillary chorioretinal atrophy may be an early glaucomatous finding in some patients with ocular hypertension. PMID- 9400784 TI - Effect of cataract extraction on the results of automated perimetry in glaucoma. AB - OBJECTIVE: To investigate the effect of cataract extraction on the results of automated perimetry in persons with glaucomatous visual field loss. SUBJECTS: Subjects from a retrospective study of visual field progression who underwent cataract extraction during follow-up were identified. Subjects came from the glaucoma service of a hospital-based tertiary referral center. METHODS: Subjects had at least 7 Humphrey 24-2 or 30-2 visual fields over 5 years or more, with an abnormal glaucoma hemifield test result on the first 2 examinations. Visual field data were transferred to a microcomputer and comparison of the visual fields immediately before and after cataract extraction was performed. RESULTS: Sixty five eyes of 50 subjects (mean age, 71.8 years) were included in the analysis. A mean improvement in mean deviation (MD) of 1.68 dB (P<.001), and a mean worsening in corrected pattern SD (CPSD) of 0.54 dB (P=.09) was observed. The mean unweighted change in threshold in the 52 points of program 24-2 was 1.58 dB, corresponding to a 43.9% increase in sensitivity. A significant correlation between improvement in visual acuity and improvement in MD was also found. A mean increase in CPSD of 1.61 dB (P=.005) occurred in subjects with dense scotomas (minimum threshold value < or = 5 dB) and preoperative CPSD of 8 dB or less. CONCLUSIONS: In persons with glaucomatous visual field defects, cataract extraction produces only a modest improvement in MD. After cataract surgery, the CPSD index worsened in many subjects with dense scotomas. This suggests that the development of cataract can mask progressive glaucomatous visual field loss in such persons. PMID- 9400785 TI - Factors associated with intraocular pressure-induced acute visual field depression. AB - OBJECTIVE: To determine the factors associated with visual field depression produced by artificial elevation of intraocular pressure (IOP). METHODS: The visual threshold was determined at 26 locations in the central visual field at a spontaneous IOP, at 30 mm Hg, at 40 mm Hg, and at the IOP immediately following release of the suction cup used to elevate the IOP artificially in 33 subjects with and without glaucoma. The net decrease in threshold sensitivity at each IOP level relative to sensitivity obtained at the spontaneous IOP was calculated (acute visual field depression). RESULTS: Factors potentially influencing the acute visual field depression between subjects were determined with stepwise regression. The reciprocal of ocular perfusion pressure, a clinical measure, was strongly correlated with acute visual field depression (dependent variable), particularly at 40 mm Hg (at 30 mm Hg, r=0.412, P=.02, n=32; and at 40 mm Hg, r=0.813, P<.001, n=33). When a second variable, the diagnosis of glaucoma, was included in the regression at 40 mm Hg, it contributed significantly (partial r=0.650, P<.001, n=26). The degree of glaucomatous damage (vertical cup-disc ratio or baseline Humphrey 24-2 visual field mean deviation) failed to correlate with acute field depression, with or without correction for ocular perfusion pressure. CONCLUSIONS: The elevation of IOP produces acute, reversible visual field depression. This depression is largely dependent on the subject's ocular perfusion pressure. The degree of depression is greater in those with glaucoma but is not strictly related to the degree of glaucomatous damage. PMID- 9400786 TI - MSL-109 adjuvant therapy for cytomegalovirus retinitis in patients with acquired immunodeficiency syndrome: the Monoclonal Antibody Cytomegalovirus Retinitis Trial. The Studies of Ocular Complications of AIDS Research Group. AIDS Clinical Trials Group. AB - OBJECTIVE: To evaluate the efficacy and safety of an intravenous human monoclonal antibody to cytomegalovirus (CMV), MSL-109, as adjuvant treatment for CMV retinitis. METHODS: Two hundred nine patients with acquired immunodeficiency syndrome and active CMV retinitis were enrolled in a multicenter, phase 2/3, randomized, placebo-controlled clinical trial. Patients received adjuvant treatment with MSL-109, 60 mg intravenously every 2 weeks, or placebo. Randomization was stratified on the basis of whether patients had untreated or relapsed retinitis. Primary drug therapy for CMV retinitis was determined by the treating physician. RESULTS: The rates of retinitis progression, as evaluated in a masked fashion, were 3.04/person-year in the MSL-109-treated group and 3.05/person-year in the placebo-treated group (P=.98; Wald test); the median times to progression were 67 days in the MSL-109-treated group and 65 days in the placebo-treated group. No differences between the 2 groups were noted in the rates of increase in retinal area involved by CMV, visual field loss, or visual acuity outcomes. The mortality rate in the MSL-109-treated group was 0.68/person year, and in the placebo-treated group, 0.31/person-year (P=.01). The mortality difference was not explained by differences in baseline variables or in concurrent antiretroviral therapy. Among patients with newly diagnosed retinitis, mortality rates were similar (MSL-109, 0.41/person-year; placebo, 0.42/person year; P=.95), whereas among patients with relapsed retinitis the MSL-109-treated group had a greater mortality rate (MSL-109, 0.83/person-year; placebo, 0.24/person-year; P=.003). However, the mortality rate in the placebo-treated patients with relapsed CMV retinitis was lower than that in the placebo-treated patients with newly diagnosed CMV retinitis and lower than that in other trials of patients with relapsed CMV retinitis. CONCLUSIONS: Intravenous MSL-109, 60 mg every 2 weeks, appeared to be ineffective adjuvant therapy for CMV retinitis. The mortality rate was higher in the MSL-109-treated group, but the reasons for this difference remain uncertain. PMID- 9400787 TI - Factors predictive of growth and treatment of small choroidal melanoma: COMS Report No. 5. The Collaborative Ocular Melanoma Study Group. AB - OBJECTIVES: To describe time to tumor growth of a prospectively followed group of patients with small choroidal melanoma and to determine baseline clinical and photographic characteristics associated with time to growth. METHODS: The Collaborative Ocular Melanoma Study (COMS) is a set of clinical trials designed to compare radiotherapy and enucleation in the treatment of medium- and large size choroidal melanoma. From December 1986 to August 1989, patients with small choroidal melanoma, not large enough to be eligible for the COMS clinical trials, were offered participation in a nonrandomized prospective follow-up study. Small choroidal melanomas were defined as 1.0 to 3.0 mm in apical height and 5.0 to 16.0 mm in largest basal dimension. A total of 204 patients were enrolled in the study and were followed up annually through August 1989. An assessment of current size of tumor, treatment status, and vital status was conducted in 1993-1994; an additional assessment of treatment and vital status was performed in 1995-1996. RESULTS: Of 188 small tumors not treated at the time of study enrollment, 46 grew during follow-up to a size that was large enough to be eligible for the COMS clinical trials. The Kaplan-Meier estimates of proportion of tumors that grew were 21% (95% confidence interval, 14%-27%) by 2 years and 31% (95% confidence interval, 23%-39%) by 5 years. Factors significantly associated with time to growth in a Cox proportional hazards regression model were greater initial tumor thickness and diameter, presence of orange pigment, absence of drusen, and absence of areas of retinal pigment epithelial changes adjacent to the tumor. CONCLUSIONS: Of small choroidal melanomas initially managed by observation, 21% demonstrated growth by 2 years and 31% by 5 years. The clinical and photographic features of these tumors confirm previous findings and are useful in identifying patients with small tumors at highest risk of short-term growth. PMID- 9400788 TI - Visual function 5 years after optic neuritis: experience of the Optic Neuritis Treatment Trial. The Optic Neuritis Study Group. AB - OBJECTIVE: To assess the 5-year visual course, including the incidence of recurrent optic neuritis, in 454 patients enrolled in the Optic Neuritis Treatment Trial. METHODS: Five-year follow-up vision testing, which included measures of visual acuity, contrast sensitivity, visual field, and color vision, was completed for 397 (87%) of the 454 patients. RESULTS: Visual function test results in the eyes that experienced optic neuritis at study enrollment (affected eyes) were normal or only slightly abnormal after 5 years in most patients; the results did not significantly differ by treatment group (P=.37 for visual acuity). The visual acuity in the affected eyes was 20/25 or better in 87%, 20/25 to 20/40 in 7%, 20/50 to 20/190 in 3%, and 20/200 or worse in 3%. The recurrence of optic neuritis in either eye occurred in 28% of the patients and was more frequent in patients with multiple sclerosis (P=.001) and in patients without multiple sclerosis who were in the prednisone treatment group (P=.004). Most eyes with a recurrence retained normal or almost normal visual function. CONCLUSIONS: Most patients retained good to excellent vision in the 5 years following an attack of optic neuritis, even if the optic neuritis recurred. Recurrences were more frequent in patients with multiple sclerosis and in those treated with oral prednisone alone. The completion of the 5-year follow-up by the Optic Neuritis Treatment Trial cohort has not altered our management recommendations based on the results we reported earlier. PMID- 9400789 TI - Systemic hyperoxia decreases vascular endothelial growth factor gene expression in ischemic primate retina. AB - OBJECTIVES: To determine whether systemic hyperoxia can reverse retinal hypoxia and decrease vascular endothelial growth factor (VEGF) gene expression in ischemic nonhuman primate retina. METHODS: Six eyes of 3 cynomolgus monkeys were studied. Retinal ischemia was induced via laser vein occlusion and confirmed with fluorescein angiography. Animals were randomly assigned to treatment with either 21% or 100% inhaled oxygen. Arterial PO2 was monitored while systemic acid-base status was maintained. An oxygen microelectrode on a micromanipulator was used to measure preretinal oxygen concentrations in ischemic and nonischemic retina in situ. RNA was isolated from fresh whole retinas, and VEGF messenger RNA levels were quantified with Northern blotting. RESULTS: The preretinal PO2 in ischemic retina was less than the PO2 in nonischemic retina in animals breathing 21% oxygen (intervascular zone PO2, 14.3+/-0.53 vs 21.8+/-0.55 mm Hg; P=.002). After 8 hours of systemic hyperoxia (arterial PO2, 512+/-18 mm Hg), the preretinal PO2 in ischemic retina increased to 166.2+/-15.6 mm Hg (21.8% oxygen) and retinal VEGF messenger RNA levels were reduced by an average of 55%. CONCLUSIONS: These data demonstrate that systemic hyperoxia can lower retinal VEGF gene expression and reoxygenate ischemic adult primate retina. PMID- 9400790 TI - Apoptosis in patients with posterior uveitis. AB - BACKGROUND: Apoptosis plays a part in the pathogenesis of autoimmune diseases. OBJECTIVE: To investigate the expression of apoptotic markers in the eyes of patients with uveitis. METHODS: With the use of immunohistochemical and in situ apoptotic detection techniques, apoptotic molecules (Fas or Fas ligand [FasL]) and nuclear DNA fragmentation were examined in 8 enucleated eyes with Behcet's disease (1), sarcoidosis (1), subretinal fibrosis and uveitis (1), sympathetic ophthalmia (4), and the Vogt-Koyanagi-Harada syndrome (1); in 5 chorioretinal biopsy specimens with acute retinal necrosis (2), multifocal choroiditis (1), sarcoidosis (1), and subretinal fibrosis and uveitis (1); and in 3 normal control eyes. RESULTS: Fas and FasL were constitutively expressed in the normal human retina, but they were expressed much less in the choroid. Increased expression of Fas and FasL was found in the retina, chorioretinal scar, and choroidal granulomas in uveitic eyes. However, Fas and FasL expression was absent in the biopsy specimens with acute retinal necrosis, and little Fas or FasL was noted on infiltrating lymphocytes. DNA fragmentation was also identified in eyes with chorioretinal scar and gliosis. CONCLUSIONS: Apoptosis occurs in uveitic eyes and may play a regulatory role in limiting ocular inflammation. In uveitic eyes, a dysregulation of the Fas-FasL apoptotic pathway may lead to gliosis and fibrosis. PMID- 9400791 TI - Sorsby fundus dystrophy: reevaluation of variable expressivity in patients carrying a TIMP3 founder mutation. AB - Interfamilial phenotypic variations in Sorsby fundus dystrophy (SFD) have given rise to controversy as to whether SFD constitutes more than 1 nosologic entity. The recent identification of the tissue inhibitor of metalloproteinases-3 (TIMP3) as the gene causing SFD has made it possible to readdress the question of genetic and clinical heterogeneity. In this study, we have extended previous findings on a Ser181Cys founder mutation in SFD families from the British Isles and show that carriers of this mutation residing in Canada, the United States, and South Africa likewise are descendants of the British ancestor. In addition, we have reevaluated the question of variable SFD phenotypes by analyzing the available clinical data on carriers of the Ser181Cys mutation. PMID- 9400792 TI - Associations with intraocular pressure in the Barbados Eye Study. AB - OBJECTIVE: To evaluate the demographic, medical, ocular, familial, and other factors possibly associated with intraocular pressure (IOP) in a black population, after excluding persons with any type of glaucoma. DESIGN: The Barbados Eye Study was a population-based study of a random sample of residents of Barbados, West Indies, aged 40 to 84 years. PARTICIPANTS: A subset of the Barbados Eye Study population consisting of 3752 black Barbados Eye Study participants without glaucoma. DATA COLLECTION: A standardized protocol included applanation tonometry and other ocular data, blood pressure measurements, anthropometry, complexion pigmentation gradings, and a comprehensive interview. MAIN OUTCOME MEASURE: Intraocular pressure was based on the average of 3 measurements at the Barbados Eye Study visit. Multiple linear regression was used to evaluate factors associated with IOP. RESULTS: Systolic blood pressure (or hypertension), diabetes history, and age were the major factors positively associated with IOP (P<.01). Other positively related factors were female gender, darker complexion, pulse rate, higher body mass, seasonality, family history of glaucoma, current alcohol use, and current smoking. These factors explained 10% of the variation in IOP. CONCLUSIONS: By identifying risk factors, these results define specific subgroups most likely to have an elevated IOP. The high IOP in this population may be linked to the high prevalence of hypertension and diabetes. Aside from age and a family history of glaucoma, none of the risk factors for high IOP evaluated in this study was similar to those associated with open-angle glaucoma. PMID- 9400793 TI - Small choroidal melanoma. PMID- 9400794 TI - Maximal medical therapy for glaucoma. PMID- 9400795 TI - Melanogenic neuroectodermal tumor of the retina (primary malignant melanoma of the retina). AB - A 35-month-old girl with leukocoria was clinically diagnosed with unilateral sporadic retinoblastoma. Macroscopic examination of her enucleated eye disclosed a white retinal tumor that appeared to be a retinoblastoma. Histopathologic examination, however, revealed that the tumor was composed of poorly differentiated neuroblastic cells, larger spindle-shaped cells, and anaplastic epithelioid cells, which is inconsistent with retinoblastoma. Immunohistochemical testing disclosed that the tumor cells were immunoreactive for melanoma-specific antigen HMB-45, while electron microscopy showed premelanosomes in the tumor cells, both of which are consistent with melanogenesis. To our knowledge, such an ocular tumor has not been reported previously. PMID- 9400796 TI - Primary basal cell carcinoma of the caruncle. AB - Basal cell carcinomas (BCCs) of the caruncle are rare. We report a case of a primary BCC of the caruncle in a 74-year-old man who was seen with a medial interpalpebral lesion. He had a history of sun exposure and multiple malignant neoplasms of the skin. The lesion was excised and histological examination showed a BCC of the caruncle. The clinical history, examination findings, and histological features are given. PMID- 9400797 TI - Myoepithelioma of the lacrimal gland. AB - A right orbital tumor was excised from a 76-year-old woman. Pathological examination showed that the tumor was composed of spindle to cuboidal cells arranged in a solid to trabecular pattern. Immunohistochemical stains were positive for S-100 protein, muscle-specific actin, cytokeratins MAK6 and AE1,3, and glial fibrillary acid protein and negative for CD34 in tumor cells. Ultrastructural features of tumor cells included microvillous processes, intercellular junctions, and intracytoplasmic filaments with electron densities. To our knowledge, this is the first non-spindle cell myoepithelioma noted to arise in the lacrimal gland. This tumor likely has a similar biological behavior to pleomorphic adenoma (benign mixed tumor). PMID- 9400798 TI - Pseudomembranous conjunctivitis following topical gentamicin therapy. PMID- 9400799 TI - Bilateral corneal infection as a complication of laser in situ keratomileusis. PMID- 9400800 TI - Hypertensive retinopathy simulating Leber idiopathic stellate neuroretinitis. PMID- 9400801 TI - Inflammatory mass of the optic nerve head associated with systemic Bartonella henselae infection. PMID- 9400802 TI - Epiretinal membrane delamination with a diamond knife. AB - Removal of membranes from the retinal surface is an integral part of many vitreoretinal surgical procedures. Two-handed tissue delamination with a knife allows precise and accurate control of dissection planes. A newly designed diamond knife provides the consistently sharp cutting edge required for this technique. PMID- 9400803 TI - Conjunctival squamous cell carcinoma treated with topical 5-fluorouracil. PMID- 9400804 TI - Familial trichomegaly. PMID- 9400805 TI - What is the evidence supporting chemotherapy for intraocular retinoblastoma? PMID- 9400806 TI - A case of primary choroidal melanoma in a patient with previous cutaneous melanoma. PMID- 9400807 TI - Should we patch corneal erosions? PMID- 9400808 TI - Women in ophthalmology. PMID- 9400809 TI - Natural killer cells: endothelial interactions, migration, and target cell recognition. AB - Natural killer (NK) cells form a unique third group of lymphocytes that differs from T and B cells in surface phenotype, target recognition, and function. By producing cytokines and exerting cytotoxicity, NK cells participate in the resistance against microbial infections and malignant disease. The research on the molecular mechanisms of migration and target cell recognition by NK cells has recently developed rapidly. NK cells express a number of adhesion molecules common to hematopoietic lineage, bind to endothelium, extravasate, and respond to chemotactic stimuli, much resembling T cells in those respects. However, NK cells are probably capable of transmigration and infiltration merely through activation by cytokines and chemokines, as opposed to the requirement of antigen presentation in the initial activation of T cells. Target cell recognition and ensuing cytotoxicity of NK cells is a sum effect of a delicate balance between the effects of inhibitory and activating NK cell receptors. NK cells express several well-defined MHC I-recognizing receptors that inactivate their functions. In pathological alterations of MHC I expression, the inhibitory receptors do not engage and thus permit the lysis of the target cell. The receptors that trigger the cytolytic machinery of NK cells are less well known. Some candidate triggering receptors have been identified and it seems that NK cell triggering is mediated by multiple receptors, as is the inhibition of cytotoxicity. For example, NK cells clearly detect target cell-bound antibodies and thus mediate antibody-dependent cytotoxicity. They may also detect carbohydrate moieties, normal but pathologically distributed adhesion molecules, as well as ligands for a number of co-stimulatory receptors. PMID- 9400810 TI - Elimination, blocking, and activation of macrophages: three of a kind? AB - In mammals, macrophages are multifunctional cells. Apart from their scavenger role in the clearance of non-self materials such as microorganisms and altered self materials such as apoptotic cells, senescent erythrocytes, immune complexes, and inflammatory products, they play a crucial role in the regulation of both innate and acquired immunity. Whereas the former activity is based on phagocytosis and intracellular degradation, the latter activity largely depends on the production and secretion of a panel of regulatory molecules such as cytokines, chemokines, and nitrogen oxide (NO). Depletion of macrophages and blocking of phagocytosis form important approaches to study the role of these cells in various host defense mechanisms. Moreover, the efficacy of drug- and gene-targeting, based on the application of particulate carrier devices, can be improved in this way. However, compounds originally described as efficacious blockers of phagocytosis simultaneously activate their production of cytokines and NO. Moreover, elimination, blocking, as well as activation of macrophages are all dependent on the concentration of such compounds. When administered in vivo, they will reach some macrophages in a high and others in a low concentration. As a consequence, the former cells may be eliminated or blocked, whereas the latter are activated by the same treatment. In this review, the various methods for suppression of macrophage functions are compared and requirements for the development of new, selective, and organ-specific macrophage-suppressing devices are discussed. PMID- 9400812 TI - Effect of alterations of blood cholesterol levels on macrophages in the myocardium of New Zealand White rabbits. AB - We investigated the effect of alterations of blood cholesterol levels on macrophages (mphi) in the myocardium of New Zealand White (NZW) rabbits. Three groups of NZW rabbits were used: controls, rabbits fed a 0.5% cholesterol enriched diet (CH-D) for 96 days, and rabbits fed a 0.5% CH-D for 96 days followed by normal chow for 4 months. Immunohistochemical analysis by mAbs directed against mphi (RAM-11) and Mn superoxide dismutase (MnSOD) were quantified by computer-assisted morphometry. Using cultured human and rabbit mphi, a cross-reaction of the human MnSOD mAbs was found as well as the predominant localization of MnSOD-immunoreactivity (IR) in mitochondria. In group 1, only a very few RAM-11-immunoreactive (ir) mphi occurred in the interstitial space of the myocardium. In group II blood cholesterol levels significantly increased in parallel with the numbers of mphi, which often contained lipid droplets (foam cells). Although blood cholesterol concentrations regressed about 10-fold in group III, mphi in the myocardium were found to be reduced only about 20%. Most mphi were also MnSOD-ir. In atherosclerotic coronary arteries RAM-11-IR was located in mphi and also extracellularly, whereas MnSOD-IR was found only in mphi. Drastically induced MnSOD in the mitochondria of mphi is suggested as an indicator of increased oxidative stress caused by in vitro conditions or by phagocytosis of low-density lipoprotein in vivo. Elevation of the cholesterol level leads to a long-term increase and its regression results in a delayed reduction of such mphi, which seem to play a key role in the atherogenesis of the coronary arteries as well. PMID- 9400811 TI - Immunomodulatory action of G-CSF in a rat model of endotoxin-induced liver injury: an intravital microscopic analysis of Kupffer cell and leukocyte response. AB - In contrast to the anticipation that in sepsis granulocyte colony-stimulating factor (G-CSF) would overactivate the nonspecific immune system by recruiting and priming leukocytes with consequent aggravation of inflammatory tissue lesions, recombinant (r) G-CSF pretreatment was protective in various experimental non neutropenic models of inflammation. The mechanisms of protection, however, are not fully understood. Using intravital fluorescence microscopy, we show that rG CSF enhances leukocyte endothelial cell interaction within the microvasculature of normal rat livers, whereas rG-CSF pretreatment of animals exposed to lipopolysaccharide (LPS) attenuates the LPS-induced leukocytic response, including stasis in sinusoids as well as rolling and adherence in postsinusoidal venules with subsequent tissue infiltration. Moreover, rG-CSF, which did not affect Kupffer cell activity in normal rat livers, reduced the immediate activation of Kupffer cells on LPS exposure, as indicated in vivo by the delayed adherence/phagocytosis of intra-arterially administered latex particles associated with attenuation of proinflammatory cytokine release (tumor necrosis factor alpha and interleukin-6). Finally, rG-CSF reduced LPS-induced nutritive perfusion failure and hepatocellular excretory dysfunction. This study provides evidence for a distinct, possibly tumor necrosis factor alpha-dependent modulation of LPS-induced cellular response within the liver by rG-CSF, thereby achieving protection against microcirculatory perfusion failure and hepatic dysfunction. PMID- 9400813 TI - Innate resistance to Listeria monocytogenes in tumor-bearing mice. AB - We have previously described the isolation, cloning, and characterization of a tumorigenic murine fibrosarcoma, designated JBS. Growth of JBS tumors in syngeneic mice initiates an anti-tumor immune response that initially manifests as progressive splenic hyperplasia and an increased proliferative ability in cultured splenocytes. In animals with tumors progressing beyond the 2 cm stage there is a reduction in spleen size and a gradual decrease in splenocyte proliferative abilities, leading to anergy at heavy tumor burdens (>3.5 cm). During the phase of immune hyperresponsiveness in tumor-bearing mice clearance of Listeria monocytogenes by components of the innate immune system is increased. This heightened resistance to infection is most likely macrophage-mediated because these mice demonstrate an increased ability to recruit macrophages to the peritoneal cavity during Listeria infection. In addition, these macrophages are highly activated in vivo as evidenced by an elevated capacity to express class II MHC (Ia) molecules. This increase in macrophage activation status is coincident with an increased capacity of splenocytes from tumor-bearing mice to secrete IFN gamma. In mice with much heavier tumor burdens (>3.5 cm), down-regulation of the immune response leads to a reduction in peritoneal macrophage numbers, decreased macrophage Ia expression, and diminished splenic clearance of L. monocytogenes. Our data demonstrate that activation of macrophages distal to the tumor site occurs as an initial consequence of tumor growth. It is only in mice with very heavy tumor burdens that functionality of macrophages is sufficiently suppressed to allow increased splenic growth of L. monocytogenes. PMID- 9400814 TI - Effects of acute ethanol exposure on cellular immune responses in a murine model of thermal injury. AB - To test the effects of acute ethanol exposure on immune function after thermal injury, mice with blood alcohol levels of 100 mg/dL were given a 15% total body surface area dorsal scald or sham injury. Bacterial challenge resulted in 100 and 40% mortality in burn + ethanol- and burn + vehicle-treated mice, respectively. Delayed-type hypersensitivity responses were also significantly suppressed in burn + ethanol-treated mice. At 1 and 4 days post-burn, concanavalin A (ConA) induced total splenocyte proliferation in burn + ethanol-treated groups was significantly decreased (P < 0.01) compared with burn + vehicle- or sham-treated animals. This decrease was not observed in total splenocytes cultured with anti CD3epsilon or among adherence-depleted splenocytes given ConA or anti-CD3epsilon. FACS analyses revealed no changes in splenocyte sub-type ratios in burn + ethanol mice. The data herein demonstrate that acute ethanol exposure before thermal injury results in enhanced susceptibility to bacterial infection and markedly suppressed cellular immunity, which appears to be macrophage dependent. PMID- 9400815 TI - Rat monocytes up-regulate NKR-P1A and down-modulate CD4 and CD43 during activation in vivo: monocyte subpopulations in normal and IFN-gamma-treated rats. AB - LEW rats were treated intravenously with recombinant rat interferon-gamma (IFN gamma) for 3 days to achieve intravascular accumulation, proliferation, and activation of monocytes. Monocytes, defined by their expression of the ED1, ED9, and Ox41 antigens, were recovered from the vasculature by perfusion with PBS/EDTA, subsequently depleted of erythrocytes and granulocytes by Percoll density gradient centrifugation, and analyzed by flow cytometry and immunocytology. In untreated and control-infused specified pathogen-free (SPF) rats, lymphocytes and monocytes formed overlapping cell populations with respect to size and internal granularity. At least two intravascular monocyte subsets, probably central and marginating cells, were distinguished by their size and differential expression of CD43, CD4, CD11a, CD18, and L-selectin. It is interesting to note that a fraction of the monocytes in normal and control infused animals carried the NKR-P1A molecule. IFN-gamma treatment provoked a duplication of monocyte size and granularity. Both the number of positive monocytes and the level of expression of NKR-P1A strongly increased after IFN gamma infusion, whereas CD43 (leukosialin) and CD4 were impressively down regulated. NKR-P1A+ L-selectin+ CD43low CD4- monocytes also occur in the vasculature of rats during immune reactions in vivo. We speculate that these cells are involved in organ damage and that their number is controlled by activation-induced cell death within the vessels. PMID- 9400816 TI - Increased expression of an endopeptidase (gamma-EGE/IDE) hydrolyzing beta endorphin during differentiation and maturation of bone marrow macrophages. AB - The presence and regulated expression of peptidase activity is a powerful mechanism with the potential to terminate or alter receptor recognition, cell membrane signal transduction, and physiological responses of immune cells to exogenous opioid peptides. In this study, the expression of an endopeptidase that hydrolyzes beta-endorphin to gamma-endorphin and other peptide products was investigated during in vitro differentiation and maturation of recombinant granulocyte-macrophage colony-stimulating factor (rGM-CSF) -derived, bone marrow derived macrophages. In freshly isolated intact isolated mouse bone marrow cells the rate of beta-endorphin hydrolysis is undetectable (<0.1 nmol beta-endorphin hydrolyzed/h/10[6] cells). However, total intracellular beta-endorphin hydrolytic activity was increased significantly to 20.0 +/- 1.7 nmol/h/10(6) cells in the mature mouse macrophages derived in vitro by culture with rGM-CSF. rGM-CSF derived macrophages expressed significantly higher levels of both protein and mRNA for the major beta-endorphin endopeptidase, gamma-endorphin-generating enzyme/insulin-degrading enzyme (gamma-EGE/IDE). Moreover, this enzymatic activity appears to be responsible for cleavage of exogenous beta-endorphin by intact rGM-CSF-derived macrophages or peritoneal macrophages to generate gamma endorphin and other peptide products. PMID- 9400817 TI - Tumor site-selective localization of an adoptively transferred T cell line expressing a macrophage lectin. AB - CTLL-2 cells were transfected with an expression vector that contained cDNA of a mouse macrophage galactose/N-acetylgalactosamine-specific calcium-type lectin (MMGL) and a stable transfectant (CTL-ML) was established. These cells and mock transfectant cells (CTL-CEP) were labeled with a long-term fluorescent cell tracer, DiI. The labeled cells were intravenously injected into mice that contained established lung metastases produced by OV2944-HM-1 ovarian tumor cells. Analyses with fluorescence microscopy of a series of frozen lung sections from the recipient mice revealed that CTL-ML preferentially accumulated in the lung metastatic nodules, whereas CTL-CEP did not. Time course studies showed that the preferential accumulation was evident 3 days after adoptive transfer. We also found that OV2944-HM-1 cells bound peanut agglutinin and Vicia villosa agglutinin B4, whose sugar specificity overlaps with the specificity of MMGL. These results suggested that MMGL molecules expressed on CTLL-2 cells contributed to their selective trafficking or retention in tumor foci possibly through recognition of tumor-associated cell surface carbohydrate antigens. These results also suggested that MMGL could be used for the selective targeting of effector cells in adoptive immunotherapy. PMID- 9400818 TI - Dietary fish oil diminishes the antigen presentation activity of rat dendritic cells. AB - Rats were fed for 6 weeks on a low fat (LF) diet or on high fat diets containing safflower oil [SO; rich in n-6 polyunsaturated fatty acids (PUFAs)] or fish oil (FO; rich in n-3 PUFAs). Lymph-borne dendritic cells (L-DC) were isolated after cannulation of the thoracic duct and were used as antigen [keyhole limpet hemocyanin (KLH)]-presenting cells in an ex vivo assay that used KLH-sensitized spleen lymphocytes as the responder cells. FO feeding significantly diminished the antigen presentation activity of L-DC compared with L-DC from rats fed each of the other diets. The antigen presentation activity of L-DC from rats fed the SO diet was greater than that of L-DC from rats fed the LF diet. Feeding the FO diet significantly reduced both the proportion of CD2-positive L-DC and the level of CD2 expression on L-DC compared with feeding each of the other diets; the proportions of L-DC staining positive for CD40, CD18, CD54, CD11a, and MHC II were unaffected by diet. However, FO feeding reduced the level of expression of CD18, CD11a, MHC II, and CD54 on L-DC compared with feeding the other two diets; the level of expression of CD40 was unaffected by diet. This is the first study to report effects of dietary fatty acids on dendritic cells. The suppressive effect of FO feeding may account for some of the beneficial effects of n-3 polyunsaturated fatty acids observed in clinical settings, such as prolonged survival of grafts and diminished chronic inflammatory responses. However, such an effect may also be detrimental because host defense toward bacterial and other antigens could be compromised. PMID- 9400819 TI - Effects of inhibitors of inflammatory mediators and cytokines on eosinophil and neutrophil accumulation induced by Mycobacterium bovis bacillus Calmette-Guerin in mouse pleurisy. AB - In this work we characterize the Mycobacterium bovis bacillus Calmette-Guerin (BCG) -induced pleurisy and investigate the role of chemical mediators and cytokines in BCG-induced granulocyte accumulation at 24 h. Intrathoracic injection of BCG in C57B1/6 mice induces a biphasic inflammatory reaction with intense leukocyte accumulation at 24 h and 15 days. Neutrophils were observed in the pleural cavity at 4-24 h, mononuclear cells and eosinophils after 24 h. A new wave of mononuclear cells and neutrophils were observed after 15 days. Pretreatments with dexamethasone, BW 755C, BW A4C, WEB 2170, L-NAME, and monoclonal antibody (mAb) anti-interleukin-5 (IL-5; TRFK-5) had inhibited the eosinophil accumulation. On the other hand, only the pretreatments with dexamethasone, L-NAME, or mAb anti-tumor necrosis factor alpha (TNF-alpha; MP6 XT3) had inhibited the neutrophil influx. These results suggest the involvement of leukotrienes, platelet-activating factor, nitric oxide, and IL-5 in the eosinophil accumulation, and a role for nitric oxide and TNF-alpha in the neutrophil influx induced by BCG. PMID- 9400820 TI - Phagocytosis of gram-negative bacteria by a unique CD14-dependent mechanism. AB - THP-1-derived cell lines were stably transfected with constructs encoding glycophosphatidylinositol (GPI)-anchored or transmembrane forms of human CD14. CD14 expression was associated with enhanced phagocytosis of serum (heat inactivated)-opsonized Escherichia coli (opEc). Both the GPI-anchored and transmembrane forms of CD14 supported phagocytosis of opEc equally well. Lipopolysaccharide-binding protein (LBP) played a role in CD14-dependent phagocytosis as evidenced by inhibition of CD14-dependent phagocytosis of opEc with anti-LBP monoclonal antibody (mAb) and by enhanced phagocytosis of E. coli opsonized with purified LBP. CD14-dependent phagocytosis was inhibited by a phosphatidylinositol (PI) 3-kinase inhibitor (wortmannin) and a protein tyrosine kinase inhibitor (tyrphostin 23) but not a protein kinase C inhibitor (bisindolyl maleimide) or a divalent cation chelator (ethylenediaminetetraacetate). Anti-LBP mAb 18G4 and anti-CD14 mAb 18E12 were used to differentiate between the pathways involved in CD14-dependent phagocytosis and CD14-dependent cell activation. F(ab')2 fragments of 18G4, a mAb to LBP that does not block cell activation, inhibited ingestion of opEc by THP1-wtCD14 cells. 18E12 (an anti-CD14 mAb that does not block LPS binding to CD14 but does inhibit CD14-dependent cell activation) did not inhibit phagocytosis of LBP-opEc by THP1-wtCD14 cells. Furthermore, CD14-dependent phagocytosis was not inhibited by anti-CD18 (CR3 and CR4 beta-chain) or anti-Fcgamma receptor mAb. PMID- 9400821 TI - Effector molecules in expression of the antimicrobial activity of macrophages against Mycobacterium avium complex: roles of reactive nitrogen intermediates, reactive oxygen intermediates, and free fatty acids. AB - We studied microbicidal activities of reactive nitrogen intermediates (RNI), free fatty acids (FFA), and reactive oxygen intermediates (ROI) against Mycobacterium avium complex (MAC) and the mode of macrophage (mphi) production of these effectors. (1) Intracellular growth of MAC in murine peritoneal mphis was accelerated by scavengers for ROI or RNI and inhibitors of nitric oxide synthase or phospholipase A2, indicating roles of ROI, RNI, and FFA in mphi anti-MAC functions. (2) Acidified NaNO2-derived RNI, FFA (linolenic and arachidonic acids), and the H2O2-mediated halogenation system exhibited a significant anti MAC bactericidal activity. The combination of RNI with FFA showed a synergistic effect. However, the H2O2-halogenation system in combination with either RNI or FFA showed an antagonism. When Listeria monocytogenes (Lm) was used as a target organism, the combinations of RNI + FFA and RNI + H2O2-halogenation gave a synergistic effect, whereas FFA + H2O2-halogenation showed an antagonism in exerting bactericidal activity. In addition, when ROI generated by the xanthine oxidase-acetaldehyde system was combined with RNI, anti-Lm but not anti-MAC activity was potentiated. (3) ROI production by murine peritoneal mphis was observed immediately after contact with MAC organisms (MAC stimulation) and ceased within 2 h. FFA release was seen 1-24 h after MAC stimulation. RNI production was initiated from 3 h and increased during the first 36 h and continued at least for 4 days. These findings suggest that RNI and FFA rather than ROI are important effectors of anti-MAC functions of mphis, and the collaborating action of RNI with FFA temporarily participates in mphi-mediated killing of MAC in the relatively early phase after MAC stimulation. PMID- 9400822 TI - Modulation of human neutrophil inflammatory responses by nitric oxide: studies in unprimed and LPS-primed cells. AB - Because nitric oxide (NO) can act both as a regulatory and as a toxic molecule, we have studied N-formyl-methionyl-leucyl-phenylalanine (fMLF)-stimulated responses of human neutrophils (PMNs) during various conditions of NO modulation in unprimed and bacterial lipopolysaccharide (LPS) -primed cells. Effects of various NO modulators were assessed on stimulated superoxide (O2-) generation, granule exocytosis, homotypic aggregation, and rises in intracellular free Ca2+ ([Ca2+]i). Significant differences in the effects of various NO modulators on inflammatory responses of PMNs kept in stirred suspension versus those kept under static and/or adherent conditions, were observed. L-arginine, the physiological substrate for NO synthase (NOS), and NG-nitro-L-arginine methyl ester, an inhibitor of NOS, both caused a 40-50% inhibition of LPS-induced priming of O2- generation in PMNs in stirred suspension, but not in LPS-primed PMNs under static or adherent conditions. The NO donors, sodium nitroprusside and S-nitroso-N acetylpenicillamine, completely abrogated the LPS-induced priming of O2- generation in PMNs in suspension, while causing only a 40-50% inhibition in PMNs under static or adherent conditions. The Ca2+ ionophore, A23187, prevented the LPS-induced priming of O2- generation without affecting O2- generation in unprimed PMNs. LPS priming of PMNs induced about a twofold increase in fMLF stimulated homotypic aggregation, exocytosis of secondary granules, and rises in [Ca2+]i. In related studies, we also provide definitive evidence for enzymatic formation of NO in human PMNs and demonstrate a significant decrease in NO levels in LPS-primed PMNs. Taken together, these findings indicate that NO modulates PMN inflammatory responses and plays a protective role in priming and activation processes of inflammatory PMNs. PMID- 9400823 TI - Rapid induction of neutrophil apoptosis by sulfasalazine: implications of reactive oxygen species in the apoptotic process. AB - Accumulating evidence indicates that neutrophils are crucially involved in the pathogenesis of inflammatory bowel diseases and rheumatoid arthritis. We therefore investigated the effect of sulfasalazine (SSZ), which is widely used in the treatment of these diseases, on neutrophil apoptosis in vitro. The apoptosis of neutrophils was determined by morphology, a DNA histogram of propidium iodide stained nuclei, and DNA fragmentation. SSZ rapidly accelerated the rate of spontaneous neutrophil apoptosis within clinically relevant concentrations. This effect is unique to neutrophils because other types of leukocytes and a number of leukocyte cell lines are resistant to SSZ. Neutrophil apoptosis caused by SSZ was abrogated by a tyrosine kinase inhibitor, a protein kinase A inhibitor, and antioxidants. The subsequent results provided pharmacological evidence that the phosphorylation of tyrosine kinase and protein kinase A and generation of reactive oxygen species are involved in SSZ-mediated neutrophil apoptosis. These data suggest that SSZ-induced neutrophil apoptosis may account, in part, for the clinical benefits of SSZ on inflammatory bowel diseases and rheumatoid arthritis. PMID- 9400824 TI - Anti-inflammatory action of dapsone: inhibition of neutrophil adherence is associated with inhibition of chemoattractant-induced signal transduction. AB - Dapsone has clinical utility as an anti-inflammatory agent but the mechanism of this action remains unknown. We have previously reported that dapsone inhibits beta2 integrin (CD11b/CD18)-mediated adherence of human neutrophils in vitro and now describe studies designed to discover how dapsone-mediated inhibition of this neutrophil function occurs. Results indicate that dapsone interferes with the activation or function of the G-protein (Gi type) that initiates the signal transduction cascade common to chemotactic stimuli. They also show that dapsone mediated suppression of this pathway inhibits the generation of second messengers essential to the activation of beta2 integrin molecules, as well as respiratory and secretory functions of neutrophils exposed to chemoattractants. We propose that the inhibition of chemoattractant-induced signal transduction by dapsone suppresses neutrophil recruitment and local production of toxic respiratory and secretory products in the affected skin of dermatitis herpetiformis and other neutrophilic dermatoses. PMID- 9400825 TI - Maintenance and down-regulation of primed neutrophil chemiluminescence activity in human whole blood. AB - Priming of polymorphonuclear neutrophils (PMN) in whole blood (by tumor necrosis factor alpha and interleukin-8 for enhancement of luminol-dependent chemiluminescence induced by human complement-opsonized zymosan) was stable for 120 min. In contrast, priming of isolated PMN in plasma-free suspension for responses to opsonized zymosan, formyl-methionyl-leucyl-phenylalanine, and phorbol myristate acetate was markedly less stable. Decay of priming was not due to irreversible inactivation of the terminal CL production machinery because PMN could be reprimed by platelet-activating factor or leukotriene B4. The tumor necrosis factor-alpha-primed state of isolated PMN was stabilized by host plasma in a concentration-dependent fashion. We conclude that PMN priming results in a dynamic state that is reversible. Our findings suggest the existence of blood borne components that may act to stabilize or modify PMN priming. PMID- 9400826 TI - Induction and modulation of macrophage Ia antigen expression by platelet activating factor. AB - Expression of major histocompatibility complex class II molecules, Ia, can be significantly augmented by interferon-gamma (IFN-gamma) in macrophages. In this study we demonstrate that platelet-activating factor (PAF) was also a potent inducer of Ia antigen expression on macrophages. PAF-induced Ia expression was both time- and dose-dependent. Maximal Ia expression was induced with 25 nM PAF after 3-h exposure to PAF. Ia expression in macrophages stimulated with PAF for 24 h was not significantly greater than unstimulated macrophages. Treatment of macrophages with IFN-gamma and PAF did not affect either the kinetics or concentration required for maximal Ia expression induced by either IFN-gamma or PAF. PAF-induced Ia expression was inhibited by the specific PAF receptor antagonists, WEB 2086, Ro 24-0238, and Ro 24-4637, indicating a receptor-mediated event. Like IFN-gamma-induced Ia expression, PAF activity was inhibited by prostaglandin E2 (PGE2). However, that expression was only inhibited after 24 h when macrophages were treated with the PGE2 synthesis inhibitors, flurbiprofen and indomethacin. These findings demonstrate that PAF, along with its role as a potent proinflammatory mediator, was also capable of inducing Ia expression on macrophages through the PAF receptor and that expression was altered by PGE2. PMID- 9400827 TI - Epithelioid cells from foreign-body granuloma selectively express the calcium binding protein MRP-14, a novel down-regulatory molecule of macrophage activation. AB - The S-100 proteins MRP-8 and MRP-14 are expressed by cells of the myelomonocytic lineage, either alone or simultaneously during certain stages of cellular differentiation. We demonstrate that MRP-14 but not MRP-8 was detected by immunostaining in the cytoplasm of epithelioid cells on the surface of round coverslips implanted for 14 days into the subcutaneous tissue of mice. Using this experimental model, our laboratory has previously shown that epithelioid macrophages are poor phagocytic cells that release a macrophage-deactivating factor (MDF) in short-term cultures. The full chemical characterization of MDF has not been achieved so far. We provide evidence that the calcium-binding protein MRP-14 was also released by epithelioid macrophages in short-term cultures and that its neutralization from the culture medium after addition of monoclonal antibody anti-MRP-14 abolished the MDF activity of the conditioned medium. Purified or recombinant human MRP-14 but not MRP-8 inhibits the respiratory burst of BCG-activated macrophages. Recombinant mouse MRP-14 also down-regulate macrophage activation in vitro, and PMA does not revert the inhibitory effect induced by MRP-14. It is thus concluded that epithelioid cells from foreign-body granuloma express and release MRP-14 in short-term cultures and that this molecule is endowed with MDF activity. PMID- 9400828 TI - Bacterial LPS and IFN-gamma trigger the tyrosine phosphorylation of vav in macrophages: evidence for involvement of the hck tyrosine kinase. AB - We and others have previously reported that tyrosine kinases play key roles in the activation of macrophages by both bacterial lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma). However, little is known regarding the substrates of tyrosine phosphorylation that mediate macrophage activation and the resultant production of inflammatory mediators. In lymphocytes and other hematopoietic lineages, tyrosine phosphorylation of the proto-oncogene vav appears to be an essential component of cell activation. In this study, we demonstrate that both LPS and rIFN-gamma trigger the prompt, dose-dependent tyrosine phosphorylation of vav in murine RAW 264.7 macrophages. In addition, vav is physically associated with the src-related kinase hck in murine macrophages, and antisense oligonucleotides specific for murine hck block both LPS and rIFN-gamma-mediated vav phosphorylation. These findings suggest that hck probably mediates vav tyrosine phosphorylation during macrophage activation and that LPS and rIFN-gamma mediated signaling pathways partially overlap. PMID- 9400829 TI - PGG-Glucan activates NF-kappaB-like and NF-IL-6-like transcription factor complexes in a murine monocytic cell line. AB - PGG-Glucan (Betafectin) is a novel soluble beta-glucan immunomodulator that enhances leukocyte microbicidal activities without inducing inflammatory cytokines. Although several different receptors for soluble and particulate beta glucans have been described, the signal transduction pathway(s) used by soluble beta-glucans have not been elucidated. We report that in a murine monocytic cell line (BMC2.3) PGG-Glucan activates nuclear factor-kappaB (NF-kappaB)-like and NF interleukin-6 (IL-6)-like transcription factors. Electrophoretic mobility shift assays showed that PGG-Glucan activation of the factors is time- and concentration-dependent. The NF-kappaB-like complex includes subunit p65 (rel-A) as one of its components, but apparently not p50 (kappaB1), p52 (kappaB2), p68 (rel-B), or p75 (C-rel) family members. The NF-IL-6-like complex contains subunit C/EBP-beta (NF-IL-6alpha) as one of its components, but apparently not C/EBP alpha or C/EBP-delta (NF-IL-6beta). As expected, lipopolysaccharide (LPS) activated p65/p50 NF-kappaB and C/EBP-beta NF-IL-6 complexes, increased the nuclear titer of p65 and p50 antigens, and increased cytokine (IL-1beta, tumor necrosis factor alpha) mRNA production. In contrast, PGG-Glucan increased the nuclear titer of p65, but apparently not p50, and did not induce cytokine mRNA production. These data demonstrate that PGG-Glucan utilizes signal transduction pathways different from those used by LPS. The data suggest that activation of the PGG-Glucan-stimulated factors is not sufficient to stimulate cytokine mRNA transcription. PMID- 9400830 TI - Signal transduction in Jurkat T lymphocytes. Evidence for early Ca2+ movements between the cytoplasm and the nucleoplasm in activated cells. AB - Spatial analyses of the distribution of Ca2+ in resting and activated T and B lymphocytes have shown that the bulk of increased [Ca2+]i appears to be associated with the nuclear region. These observations suggest that Ca2+ is released from the perinuclear space or that it diffuses to the nucleoplasm, or both. We have used laser scanning confocal microscopy to assess whether cytoplasmic diffusion of Ca2+ could contribute to the rise in nuclear Ca2+. We found that the activation of individual Jurkat cells by use of an anti-Ti (beta subunit) mAb induced a nucleus-associated increase in [Ca2+]i. In cells loaded with the InsP3 receptor antagonist heparin, the nuclear Ca2+ response was abolished but not the response to thapsigargin. Evidence for a cytoplasmic Ca2+ response was obtained by loading Jurkat cells with a cytoplasm-restricted Ca2+ probe (Calcium Green-1-Dextran). These observations suggested that a process of diffusion of cytoplasmic Ca2+ contributed to the rise of nuclear Ca2+ in Jurkat T cells. This interpretation was supported by the findings (1) that rapid scanning of thapsigargin-released Ca2+ showed an inverse relationship between the levels of cytoplasmic and nuclear Ca2+ and (2) that modulation of the external concentration of Ca2+ in thapsigargin-treated Jurkat cells showed a time dependent decrease of fluorescence from the nucleoplasm that was reversed by raising the concentration of external Ca2+. We conclude that Ca2+ can rapidly diffuse between the cytoplasm and the nucleoplasm in activated Jurkat T lymphocytes and that hydrophilic Ca2+ probes largely partition to the nucleoplasm, thus giving rise to distorted nucleus-to-cytoplasm fluorescence ratios. PMID- 9400831 TI - Induction of endotoxin tolerance depletes nuclear factor-kappaB and suppresses its activation in rat alveolar macrophages. AB - To investigate the mechanism of endotoxin tolerance in macrophages, a rat alveolar macrophage cell line (NR8383) was rendered endotoxin tolerant by treatment with endotoxin at 40 ng/mL for 48 h. This treatment induced a state of tolerance such that subsequent exposure to high-dose endotoxin (5 microg/mL) resulted in decreased production of macrophage inflammatory protein-2, tumor necrosis factor alpha, and nitric oxide compared to endotoxin-sensitive cells. Suppressed mediator production by endotoxin-tolerant cells was associated with impaired activation of nuclear factor-kappaB (NF-kappaB) in response to treatment with 5 microg/mL of endotoxin. This impairment of NF-kappaB activation was found to be associated with depletion of latent NF-kappaB (both RelA and p50) in the cytoplasm of endotoxin-tolerant cells. These data suggest that a mechanism of endotoxin tolerance is depletion of RelA/p50, which could limit the amount of NF kappaB available for activation by subsequent stimuli and thereby inhibit transcription of NF-kappaB-dependent genes. Limiting NF-kappaB-dependent inflammatory gene transcription by inducing endotoxin tolerance is a potential therapeutic strategy for diseases where excessive production of inflammatory mediators leads to tissue injury. PMID- 9400832 TI - The cytoplasmic domains of complement regulatory protein CD46 interact with multiple kinases in macrophages. AB - Membrane cofactor protein (CD46), which normally protects autologous cells from complement lysis, is the human cell receptor for measles virus (MV). Interaction between MV and CD46 on monocytes can lead to suppression of monocyte activation. We have investigated the interaction between the cytoplasmic sequences of CD46 and kinases in a mouse macrophage cell line. Glutathione-S-transferase (GST) fusion proteins bearing the Cyt1 or Cyt2 alternative cytoplasmic domain of CD46 associate with macrophage kinase activity, which phosphorylates multiple proteins co-purified with the GST fusion proteins. Association with the macrophage kinase activity correlates with tyrosine phosphorylation of the CD46 cytoplasmic domains. Removing the CD46 sequences or introducing a frame-shift mutation abrogates the association with macrophage kinase activity. Renaturation studies reveal multiple kinases with apparent molecular mass of 82, 79, 58, and 50/49 kDa, which associate specifically with both CD46 cytoplasmic domains. Alanine substitutions at a juxtamembrane Tyr-X-X-Leu motif in the Cyt1 domain completely abrogate the association with macrophage kinases and tyrosine phosphorylation of Cyt1; but similar substitutions in the Cyt2 domain only partially reduce the association with kinases and tyrosine phosphorylation of Cyt2. These results reveal a specific interaction between complement regulatory protein CD46 and macrophage kinases. These findings may provide an important clue for understanding immune modulation by MV. PMID- 9400833 TI - Agonist-specific tyrosine phosphorylation of Cbl in human neutrophils. AB - The effects of soluble and particulate agonists on the tyrosine phosphorylation levels of the proto-oncogene Cbl in human neutrophils were examined. Experimental conditions allowing the maintenance of Cbl as well as of its tyrosine phosphorylation status were first established. Their use allowed us to observe that Cbl was tyrosine phosphorylated in response to some (FcgammaRII ligation, opsonized bacteria and zymosan, granulocyte-macrophage colony-stimulating factor, monosodium urate, and calcium pyrophosphate microcrystals), but not all (fMet-Leu Phe, interleukin-8) neutrophil agonists. Cbl was also shown to account for a varying proportion of the 120-kDa phosphoprotein(s) observed in response to the above stimuli. These data establish that Cbl is present in human neutrophils and that its level of tyrosine phosphorylation is modulated by some of these cells' agonists, and in particular by phagocytic particles. Furthermore, the signaling pathways activated by chemotactic factors and the other neutrophil stimuli tested in this investigation diverge at or downstream from the tyrosine phosphorylation of Cbl. PMID- 9400834 TI - Monocyte chemoattractant protein-2 is a potent agonist of CCR2B. AB - The binding and functional activity of the CC chemokines monocyte chemoattractant protein-1 (MCP-1), MCP-2, and MCP-3 have been characterized using Chinese hamster ovary DXB-11 cells transfected with the chemokine receptor CCR2B. Receptor binding studies demonstrated that 125I-labeled MCP-1 bound to a single class of high-affinity receptors with a Kd of 0.14 (0.07-0.32) nM. In competition studies MCP-1, MCP-2, and MCP-3 completely inhibited 125I-labeled MCP-1 binding with Ki values of 0.3 (0.16-0.46), 8.8 (3.4-26), and 12.2 (0.6-22) nM, respectively. In calcium mobilization studies, MCP-1 and MCP-3 induced robust elevations in intracellular calcium concentrations, whereas MCP-2 was only weakly active. In contrast, using changes in extracellular acidification rate as a functional readout, all three chemokines were identified as potent agonists of CCR2B. These data demonstrate that MCP-2, in addition to MCP-1 and MCP-3, is a potent agonist of CCR2B and furthermore that MCP-2 activates either different or a subset of the signaling pathways activated by MCP-1 and MCP-3. PMID- 9400836 TI - Multiresolution, model-based segmentation of MR angiograms. AB - During the last decade, the quality of MR angiograms has risen substantially and their clinical utility has been demonstrated progressively. This acceptance has created a need for tools with which to summarize and display the information available. We have used a model-based segmentation technique to extract vascular morphology and local flow parameters from phase contrast MR angiograms. A multiresolution data structure is used as the basis of recursive decision-making to identify regions of blood flow. The resulting data representation allows more efficient data handling in subsequent processing and visualization and is directly applicable to the creation of a connected graph model of vascular regions. We describe this flow feature extraction algorithm and demonstrate the utility of the results. PMID- 9400835 TI - SPIO-enhanced 2D-TOF MR angiography of the portal venous system: results of an intraindividual comparison. AB - The purpose of this study was to investigate whether MR angiography (MRA) of the portal venous system may be improved by means of superparamagnetic iron oxides (SPIOs) during accumulation phase imaging and to study the underlying contrast mechanisms. MRA of the portal venous system was performed on 48 patients before and after intravenous injection of a new SPIO (Resovist, Schering AG, Berlin, Germany). Resovist, as a predominantly liver parenchymal darkening agent on T2 weighted MR images with uptake into the reticuloendothelial cell system, was administered intravenously by bolus injection of 8 to 12 micromol Fe/kg body weight. Patients were scanned with breath-hold coronal and axial two-dimensional (2D) time of flight (TOF) MRA (TR = 31.0 msec, TE = 9.8 msec, flip angle (FA) = 50 degrees, and 6.9-second acquisition time per section) sequences. Signal intensity values of liver parenchyma, the portal venous system, and background were obtained for quantitative analysis. The clinical relevance of additional plain and contrast-enhanced MRA studies for surgical planning was assessed by independent reading of three readers. Liver signal-to-noise ratio (SNR) significantly decreased following iv injection of Resovist; however, SNR values of the portal veins or hepatic veins did not change significantly. Visibility of the portal venous system improved significantly (tertiary branches visible: pre in 15.2% versus post in 87.0% of patients). Resovist-enhanced 2D-TOF MRA may improve planning of liver resections by better demonstrating the relationship of central liver lesions and vessels on source images. The decrease in liver SNR at a constant vessel SNR after iv injection of Resovist improves MRA of the liver. SPIO-enhanced 2D-TOF MRA scans are superior to plain 2D-TOF MRA studies and may be added for the workup of preoperative patients. PMID- 9400838 TI - Intravascular contrast agent improves magnetic resonance angiography of carotid arteries in minipigs. AB - This study was designed to optimize three-dimensional (3D) time-of-flight (TOF) magnetic resonance angiography (MRA) sequences and to determine whether contrast enhanced MRA could improve the accuracy of lumen definition in stenosed carotid arteries of minipigs. 3D TOF MRA was acquired with use of either an intravascular (n = 13) and/or an extravascular contrast agent (n = 5) administrated at 2 to 4 weeks after balloon-induced injury to a carotid artery in 16 minipigs. Vascular contrast, defined as signal intensity differences between blood vessels and muscle normalized to the signal intensity of muscle, was compared before and after the injection of each contrast agent and between the two agents. Different vascular patencies were observed among the animals, including completely occluded vessels (n = 5), stenotic vessels (n = 3), and vessels with no visible stenosis (n = 8). Superior vascular contrast improvement was observed for small arteries and veins and for large veins with the intravascular contrast agent when compared with the extravascular contrast agent. In addition, preliminary studies in two of the animals showed a good correlation for the extent of luminal stenosis defined by digital subtraction angiography compared with MRA obtained after administration of the intravascular contrast agent (R2 = .71, with a slope of .96 +/- .04 by a linear regression analysis). We concluded that use of an intravascular contrast agent optimizes 3D TOF MRA and may improve its accuracy compared with digital subtraction angiography. PMID- 9400839 TI - Use of k-space segmentation in MR velocity mapping for rapid quantification of CSF flow. AB - In this work, a k-space segmentation technique for quantitative studies of pulsatile cerebrospinal fluid (CSF) flow is suggested. Three k-space lines are sampled for each of two interleaved gradient-echo sequences (velocity-compensated and velocity-encoded) within each repetition interval. Nine cardiac phases are obtained at a heart rate of 60 bpm with maintained nominal resolution and a factor of 3 in reduction of acquisition time relative to our conventional nonsegmented flow quantification protocol. Segmented and conventional sequences were compared in phantoms, in healthy volunteers, and in two patients with clinically suspected normal pressure hydrocephalus. Good agreement between flow curves obtained with the two sequences was demonstrated in vitro as well as in vivo. A slight underestimation of flow values in volunteers was attributed to data filtering when using the segmented sequence. Because the CSF circulation is complex and tightly connected to the vascular circulation, specific clinical applications may require flow studies at multiple positions and with different velocity encoding. In such cases, the proposed sequence can be used to gain time, but alternatively, the segmentation technique can be used to further increase spatial resolution within reasonable examination times. PMID- 9400837 TI - Ultrasmall superparamagnetic iron oxide particles (AMI 227) as a blood pool contrast agent for MR angiography: experimental study in rabbits. AB - The purpose of this study was to evaluate the contribution of an ultrasmall superparamagnetic iron oxide particles (USPIOs) based contrast agent (AMI 227), in a transverse three-dimensional time-of-flight TONE MR angiography sequence of abdominal aorta in rabbits. The main goal was to assess improvement in the visualization of small arteries such as renal arteries, when using such a sequence. Imaging experiments were performed on a 1.5 T magnet, using a transverse 3D time-of-flight (TOF) tilted optimized nonsaturating excitation (TONE) sequence with magnetization transfer suppression. The contrast media used were composed of a USPIO core surrounded by a dextran-surfactant (AMI 227). Different concentrations of AMI 227 were evaluated in 12 rabbits. Concentrations varied within the range 8.5-34 micromol Fe/kg - bw: 8.5 micromol Fe/kg (three rabbits); 17 micromol Fe/kg (three rabbits); 25.5 micromol Fe/kg (three rabbits); 34 micromol Fe/kg (three rabbits). A visual analysis based on the improvement of visualization of small arteries (renal arteries) on MIP images and a quantitative analysis based on the percentage of contrast enhancement of the aorta plotted against distance in the slab from the top edge of the acquisition volume were obtained. A signal-to-noise ratio enhancement of the distal part of the aorta and only improvement in the delineation of the renal arteries were noted when using low concentrations of the contrast media. A loss of signal-to-noise ratio of the aorta and a decrease in arterial visualization were respectively noted with higher concentration of contrast media. In this experimental study, using a transverse three-dimensional TOF TONE MR angiography sequence of renal arteries, in which sequence the saturation effect is minimized, the use of AMI 227 allows only improvement in the delineation of small arteries when using low concentrations of contrast media. PMID- 9400840 TI - Coronary MR angiography during peak-systole: work in progress. AB - The purpose of this study is to compare the quality of images of coronary arteries obtained with two-dimensional breath-hold coronary MR angiography during peak systole and mid diastole. Two-dimensional coronary MR angiography was performed in eight normal volunteers at peak systole and in mid diastole with a commercial 1.5-T MR imager. An ultrafast gradient-echo sequence with incremented flip angle series and k-space segmentation was used. The image quality grade, length, and proximal diameter of each visualized coronary artery were measured. The highest quality images in systole and diastole were compared. Coronary MR angiography provided high quality images in systole and diastole in 14 of 16 coronary vessels (87.5%). In 8 of 14 vessels (57%), there was no visible coronary MR angiogram image degradation when comparing peak systolic with mid-diastolic images. In 4 of 14 vessels (29%), there was mild MR image degradation. There was significant MR image degradation in only one case (7%). And in one case (7%), there was mild image improvement during systole. The width and length of the visualized coronary vessels did not change significantly from diastole to systole. Existing two-dimensional breath-hold coronary MR angiography provides MR images during peak systole and mid-diastole with little or no perceptible difference in quality. PMID- 9400841 TI - MRI for the evaluation of regional myocardial perfusion in an experimental animal model. AB - Myocardial perfusion was assessed in nine pigs using ultrafast gradient-echo MRI (.5 T, 15-mT/m gradients) at different levels of myocardial blood flow (range, .005-1.84 ml/min/g), generated either by adenosine infusion or by a mechanical occluder, and measured independently using radiolabeled microspheres. Sixty-four consecutive, ECG-triggered, diastolic, short axis images of the left ventricle were obtained during intravenous bolus injections (n = 30) of .05 mmol/kg of gadopentetate dimeglumine. Relative changes in peak intensity, time to peak intensity, washin slope, correlation coefficient, and cross-correlation coefficient were computed from the time-intensity curves obtained from four regions of interest, namely septal, anterior, lateral, and inferior walls. The values from the inferior wall acted as reference for evaluating relative changes in the other three regions. The cross-correlation coefficient (P < .001, rho = .60) and the peak intensity (P < .001, r = .72) showed the best correlation with myocardial blood flow. The washin slope showed a weak positive trend (P < .05), but the low value of r (r = .28) indicated that the use of this parameter to predict flow was invalid; the correlation coefficient and time to peak intensity were not correlated (P = ns). In conclusion, this study shows that it is possible to evaluate relative myocardial perfusion after the first pass of a an intravenously injected bolus of gadopentetate dimeglumine, using dynamic MRI on a conventional medium field MRI system. The cross-correlation coefficient and the peak intensity resulted in more efficient parameters to evaluate relative inhomogeneity of regional myocardial perfusion. PMID- 9400842 TI - Novel application of breath-hold turbo spin-echo T2 MRI for detection of acute myocardial infarction. AB - To assess the clinical utility of the breath-hold turbo spin-echo T2-weighted MRI in patients with acute myocardial infarction, the results of MRI were compared with those of electrocardiography, coronary angiography, and thallium-201 single photon emission tomography (SPECT) in 23 patients and 5 healthy volunteers. To compare MRI and thallium-SPECT, the left ventricle was divided into five segments, and the presence of myocardial infarction was determined in each segment. MRI demonstrated an abnormally bright signal in 49 of 140 segments (five segments each from 23 patients and 5 volunteers); thallium-SPECT showed a fixed perfusion defect in 52 segments, for an 85% diagnostic concordance rate. The size of the myocardial infarction measured on MRI corresponded well to that measured on thallium-SPECT (r = .70, P < .01). Breath-hold turbo spin-echo T2 MRI can be used for detection of acute myocardial infarction in conjunction with thallium SPECT, especially when accurate localization of lesion, increased spatial resolution, and anatomic landmarks are needed. PMID- 9400843 TI - Water-fat imaging with direct phase encoding. AB - A new method is introduced for water-fat imaging. With three acquisitions, a general direct phase encoding (DPE) of the chemical shift information is achieved. Pixels containing both water and fat are solved directly. Pixels with only a single component are resolved with local and global orientation filters, which use phase information from neighboring pixels. The fact that a single component is more likely to be water than fat in living tissues is also useful. A second pass solution yields water and fat images with superior signal-to-noise ratio. Unlike other methods, DPE does not rely on the error-prone phase unwrapping; also, it easily handles disconnected tissues. Because the magnetization vectors of water and fat are sampled not only at parallel or antiparallel, they can be not only separated but also identified respectively, which is desirable for routine clinical work. DPE has been implemented on several imagers at various field strengths and has been demonstrated in a large number of clinical cases to be useful and robust in various parts of the body. PMID- 9400844 TI - Multifeature analysis of Gd-enhanced MR images of breast lesions. AB - The objective of this study was to determine whether linear discriminant analysis of different independent features of MR images of breast lesions can increase the sensitivity and specificity of this technique. For MR images of 23 benign and 20 malignant breast lesions, three independent classes of features, including characteristics of Gd-DTPA-uptake curve, boundary, and texture were evaluated. The three classes included five, four and eight features each, respectively. Discriminant analysis was applied both within and across the three classes, to find the best combination of features yielding the highest classification accuracy. The highest specificity and sensitivity of the different classes considered independently were as follows: Gd-uptake curves, 83% and 70%; boundary features, 86% and 70%; and texture, 70% and 75%, respectively. A combination of one feature each from the first two classes and age yielded a specificity of 79% and sensitivity of 90%, whereas highest figures of 93% and 95%, respectively, were obtained when a total of 10 features were combined across different classes. Statistical analysis of different independent classes of features in MR images of breast lesions can improve the classification accuracy of this technique significantly. PMID- 9400845 TI - MRI of pelvic masses: efficacy of the rectal superparamagnetic contrast agent Ferumoxsil. AB - The purpose of this study was to determine the potential value of rectally administered Ferumoxsil for pelvic MRI. Twenty patients with suspected rectosigmoid and ovarian tumors were prospectively examined before and after rectal administration of 300 to 600 ml of the superparamagnetic contrast agent Ferumoxsil. Imaging parameters (1.5-T system, phased-array coil, transverse T1 weighted spin echo (SE) and T2-weighted turbo-SE sequences, 6-mm slice thickness, 350-mm field of view [FOV], 512 x 512 matrix) were kept constant. Images were evaluated for tumor presence, lymphadenopathy, bowel involvement, and peritoneal implants. Precontrast and postcontrast studies were rated for bowel delineation, lesion/organ-to-bowel differentiation, presence of artifacts, and confidence of diagnosis. Delineation of rectum and sigmoid colon (P < .001) as well as separation of bowel and vagina, uterus/adnexa, dome of the urinary bladder, lymph nodes, and vessels improved significantly (P < .01) on both T1-weighted and T2 weighted postcontrast sequences. The rectal contrast agent did not increase the level of artifacts. Changes in diagnosis in 7 of 20 patients were mainly due to identification of colorectal tumors or bowel involvement on postcontrast images. The readers' diagnostic confidence was significantly higher on contrast-enhanced than on unenhanced studies (93 +/- 6.4% vs 68 +/- 25.2%). Rectal application of Ferumoxsil improves lesion/organ-to-bowel delineation and overall diagnostic confidence in pelvic MRI. PMID- 9400846 TI - Rapidly enhancing hepatic hemangiomas at MRI: distinction from malignancies with T2-weighted images. AB - The purpose of this study is to describe a subset of atypical hepatic hemangiomas that enhance rapidly and diffusely and to determine whether heavily T2-weighted images could distinguish between atypically enhancing liver hemangiomas and hypervascular malignancies. A retrospective search of MR records identified seven patients with liver hemangiomas that demonstrated diffuse early enhancement and 23 patients with biopsy-proven malignant liver lesions that were hypervascular on dynamic gadolinium-enhanced MR images. Quantitative analysis of signal intensity measurements was performed on the T2-weighted images, heavily T2-weighted (TE > 140), and dynamic gadolinium-enhanced images. Blinded reader comparison of the T2 weighted images and gadolinium-enhanced images was performed. Hypervascular hemangiomas enhanced to a greater degree than hypervascular malignant liver lesions on the early phase gadolinium-enhanced images. Perilesional parenchymal enhancement was demonstrated in five cases of rapidly enhancing hemangiomas. Signal intensity and contrast-to-noise ratios on the heavily T2-weighted images of the hemangiomas were significantly greater than that of the hypervascular malignant lesions (P < .05). Hemangiomas were differentiated from the hypervascular malignant liver lesions with high accuracy (97-100%) by three blinded readers based on the T2-weighted images. A subset of hemangiomas have atypical rapid diffuse enhancement on dynamic gadolinium-enhanced images. These atypical hemangiomas can be distinguished from hypervascular malignant liver lesions on T2-weighted MR images. PMID- 9400847 TI - Liver lesion detection, characterization, and effect on patient management: comparison of single-phase spiral CT and current MR techniques. AB - This study compares liver lesion detection, characterization, and effect on patient management between single-phase spiral CT and MRI using spoiled gradient echo (SGE), T2-weighted fat-suppressed spin echo, and serial post gadolinium SGE. All patients with suspected liver lesions who underwent spiral CT and MRI within a 1-month period between January 1993 and September 1996 were included in the study. Spiral CT and MRI were interpreted prospectively in a blinded fashion by separate individual experienced investigators, and lesion detection and characterization were determined. Confirmation was obtained by surgery (6 patients), biopsy (18 patients), imaging follow-up (36 patients), or combined reading of all imaging studies and clinical follow-up (29 patients). Effect on patient management was determined by combined chart review and interview of the patients' physicians and by retrospective clinical assessment performed by a surgical oncologist and medical oncologist separately. Eighty-nine patients were included in the study. Regarding true positive lesion detection, 295 and 519 lesions were detected on spiral CT and MR images, respectively, which was significantly different on a patient-by-patient basis (P < .001). More lesions were detected on MR than on spiral CT in 44 of 89 patients (49.4%), and 11 of these 44 patients had lesions shown on MRI in whom no lesions were apparent on CT images. No patients had true positive lesions shown on spiral CT that were not shown on MRI. Regarding lesion characterization, 129 and 466 lesions were characterized on spiral CT and MRI images, respectively, which was significantly different on a patient-by-patient basis (P < .001). More lesions were characterized on MR than CT images in 67 patients (75.3%). Regarding effect on patient management, chart review with physician interview demonstrated that findings on MRI provided information that altered patient management as compared with findings on spiral CT in 57 patients. Retrospective clinical evaluation by the surgical and medical oncologist showed that MRI was considered to have a greater effect on patient management than spiral CT in 58 and 55 patients, respectively. Comparing current MRI technique to single-phase spiral CT, MRI detected more lesions in 49.4% and characterized more lesions in 75.3% of patients investigated for focal liver disease. MRI had a greater effect on patient management in each of the three methods than single-phase spiral CT in more than 61% of patients. PMID- 9400848 TI - RARE imaging of PCr in human forearm muscles. AB - Rapid acquisition with relaxation enhancement (RARE) sequences have been used to map the 31P phosphocreatine (PCr) signal in human forearms at 4.7 T. Signal-to noise levels of approximately 10 were achieved from the major muscle groups in 5.5-minute scan times with a spatial resolution of 4 x 2 x 2 cm3. Exercise caused demonstrable reductions in PCr signal from activated muscles, which correlated with affected muscle groups in T2-weighted proton images. RARE imaging of the PCr signal at 4.7 T is feasible and, with technically achievable improvements in signal-to-noise ratio, should prove useful in studying energy metabolism in muscle and other organs. PMID- 9400849 TI - High resolution, short echo time sodium imaging of articular cartilage. AB - The sodium present in articular cartilage interacts with the negatively charged proteoglycan aggregates in the matrix of the cartilage. Sodium images of short echo time may be useful for detecting changes that occur in the proteoglycan content of the cartilage. Such changes are indicative of early osteoarthritic damage, for example. Using an asymmetric short echo technique, sodium images of high resolution and signal-to-noise ratio that demonstrate anatomic features of the cartilage are presented. These images were obtained with echo times as short as 1 msec, at an in-plane resolution of 39 microns by 117 microns and signal-to noise ratios of up to 40:1. PMID- 9400850 TI - A segment-interleaved motion-compensated acquisition in the steady state (SIMCAST) technique for high resolution imaging of the inner ear. AB - MRI, with ever-increasing spatial resolution, recently has depicted progressively more anatomic details of the inner ear and is playing an important role in the diagnostic evaluation of patients with sensorineural hearing loss. We present a three-dimensional (3D) segment-interleaved, motion-compensated acquisition in steady state (SIMCAST) sequence that allows further increase in spatial resolution in reasonable scan times minimizing artifacts due to susceptibility and motion. The sequence uses gradient moment nulling over TR and segmented interleaved acquisition of multiple data sets with different radiofrequency (RF) phase-cycling schemes. Combination of data from multiple acquisitions by averaging and maximum intensity projection were compared. Images of phantoms and in vivo inner ears were obtained with both full and fractional echoes and compared with other high resolution techniques such as three-dimensional gradient echo and two-dimensional (2D) and three-dimensional fast spin-echo (FSE) sequences. The new sequence achieves improved signal-to-noise ratio (SNR) and spatial resolution resulting in improved depiction of inner ear structures. PMID- 9400851 TI - Brain atrophy progression measured from registered serial MRI: validation and application to Alzheimer's disease. AB - In this paper, we validate the brain boundary shift integral (BBSI) as a measure of brain atrophy and demonstrate its application in Alzheimer's disease (AD). Nineteen normal subjects and nine patients with AD underwent serial three dimensional MRI (6- to 30-month scan intervals). Repeat studies were registered to the baseline studies. The accuracy of the BBSI was assessed by comparison with simulated atrophy and with segmentation; it was also tested for reproducibility, linearity, and its ability to discriminate patients with AD from healthy controls. The BBSI correlated closely with simulated volumes of atrophy (r = 1.000). The mean absolute difference between repeat measures was 1.51 ml or .13% of mean brain volume. Rates of atrophy from all 18 AD scan pairs were widely separated from those of all 31 control pairs. In matched subgroups, the mean (SD) annual rate of brain atrophy in nine controls was .24% (.32%), compared with 2.78% (.92%) in nine patients with AD. We conclude that the BBSI is a linear and highly reproducible measure of atrophy with potential uses in the early diagnosis and measurement of progression in Alzheimer's disease. PMID- 9400852 TI - Magnetization transfer fast imaging of implanted glioma in the rat brain at 4.7 T: interpretation using a binary spin-bath model. AB - C6 glioma cells were implanted in the left caudate nucleus of the rat brain. Histologic studies confirmed the presence of neoplastic tissue surrounded by a thin edematous region. Proton magnetization transfer contrast (MTC) fast imaging, using continuous wave off-resonance irradiation, was performed in vivo at 4.7 T with the rapid acquisition with relaxation enhancement (RARE) sequence. The observed MTC allowed very clear distinction of the tumoral region, in which magnetization transfer (MT) ratios were lower than in healthy tissues. Contrasts were analyzed as a function of the offset frequency and the amplitude of the radiofrequency (RF) irradiation. The contrast was higher between the contralateral basal ganglia and the tumor and lower between the tumor and the temporal lobe. Modeling of MT in the three brain regions was performed using a system including free water and a pool of protons with restricted motions. The rate of exchange between the two pools exhibited a decreasing hierarchy from the basal ganglia to the tumor. T2B values for the immobile protons ranged from 9.3 microsec in the basal ganglia to 7.5 microsec for the glioma. The acquisition conditions leading to the highest contrasts between the tumor and the healthy tissues correspond to 3,000 Hz offset frequency and 300 to 700 Hz RF irradiation amplitude. PMID- 9400853 TI - Dynamic imaging of intracranial lesions using fast spin-echo imaging: differentiation of brain tumors and treatment effects. AB - The purpose of this study was to develop a technique for differentiating between recurrent brain tumors and treatment-related changes, such as radiation necrosis, using dynamic MRI. Ninety-five patients with intracranial mass lesions were evaluated using T1-weighted fast spin-echo (FSE) MRI at 1.5 T. Pathologies included treatment-related changes (n = 32), primary tumors (n = 41), metastatic tumors (n = 5), meningiomas (n = 4), and mixed primary/treatment related changes (n = 13). Signal enhancement-time curves were analyzed by fitting to a sigmoidal exponential function. Maximal enhancement rates were calculated as the first derivative of the fitted curve. Based on the maximal enhancement rates, treatment related changes could be differentiated from primary tumors, metastatic tumors, and meningiomas at the P < .05 confidence level. Lesions of mixed tumor and treatment-related change had intermediate values. Dynamic MRI can be used to differentiate treatment-related changes from primary tumors in previously treated patient populations based on maximal enhancement rates. Individual case studies demonstrate the clinical significance of these findings. PMID- 9400854 TI - Fuzzy clustering of gradient-echo functional MRI in the human visual cortex. Part I: reproducibility. AB - Reproducibility of human functional MRI (fMRI) studies is essential for clinical and neuroresearch applications of this new human brain mapping method. Based on a recently presented study on reproducibility of gradient-echo fMRI in the human visual cortex (Moser et al. Magn Reson Imaging 1996; 14:567-579), comparing the performance of three different threshold strategies for correlation analysis, we demonstrate that (a) fuzzy clustering is a robust, model-independent method to extract functional information in time and space; (b) intertrial reproducibility of cortical activation is significantly improved by the capability of fuzzy clustering to separate signal contributions from larger vessels, running perpendicular to the slice orientation, from activation apparently close to the primary visual cortex; and (c) for repeated single subject studies, SDs of <20% for signal enhancement in approximately 80% of the studies and SDs of <30% for activated area size in approximately 65% of the studies are obtained. This, however, depends also on signal-to-noise ratio, (motion) artifacts, and subject cooperation. PMID- 9400855 TI - Fuzzy clustering of gradient-echo functional MRI in the human visual cortex. Part II: quantification. AB - Fuzzy cluster analysis (FCA) is a new exploratory method for analyzing fMRI data. Using simulated functional MRI (fMRI) data, the performance of FCA, as implemented in the software package Evident, was tested and a quantitative comparison with correlation analysis is presented. Furthermore, the fMRI model fit allows separation and quantification of flow and blood oxygen level dependent (BOLD) contributions in the human visual cortex. In gradient-recalled echo fMRI at 1.5 T (TR = 60 ms, TE = 42 ms, radiofrequency excitation flip angle [theta] = 10 degrees-60 degrees) total signal enhancement in the human visual cortex, ie, flow-enhanced BOLD plus inflow contributions, on average varies from 5% to 10% in or close to the visual cortex (average cerebral blood volume [CBV] = 4%) and from 100% to 20% in areas containing medium-sized vessels (ie, average CBV = 12% per voxel), respectively. Inflow enhancement, however, is restricted to intravascular space (= CBV) and increases with increasing radiofrequency (RF) flip angle, whereas BOLD contributions may be obtained from a region up to three times larger and, applying an unspoiled gradient-echo (GRE) sequence, also show a flip angle dependency with a minimum at approximately 30 degrees. This result suggests that a localized hemodynamic response from the microvasculature at 1.5 T may be extracted via fuzzy clustering. In summary, fuzzy clustering of fMRI data, as realized in the Evident software, is a robust and efficient method to (a) separate functional brain activation from noise or other sources resulting in time-dependent signal changes as proven by simulated fMRI data analysis and in vivo data from the visual cortex, and (b) allows separation of different levels of activation even if the temporal pattern is indistinguishable. Combining fuzzy cluster separation of brain activation with appropriate model calculations allows quantification of flow and (flow-enhanced) BOLD contributions in areas with different vascularization. PMID- 9400856 TI - Perfusion-weighted MRI using gadobutrol as a contrast agent in a rat stroke model. AB - The purpose of this study was to examine the new nonionic contrast agent gadobutrol in MR perfusion-weighted imaging, including the influence of different concentrations and dosages of the agent on the sensitivity to perfusion alterations. Sixteen rats were examined within 35 to 105 minutes after endovascular occlusion of the middle cerebral artery. A fast T2*-weighted fast low-angle shot (FLASH) sequence was used to acquire four images before and 16 images after bolus injection of .1, .2, .3, and .4 mmol/kg gadobutrol as .5 molar and 1.0 molar formulation. From user-defined regions, we obtained the maximum signal decrease, the relative regional cerebral blood volume, and the bolus delay. Contrast between ischemic and nonischemic regions during bolus passage increased with dose and concentration of the contrast agent. For low doses (.1 and .2 mmol/kg), the ischemic lesion could not or could barely be discerned. For higher doses (.3 and .4 mmol/kg), administration of the 1 molar contrast agent yielded a better contrast between ischemic and nonischemic tissue. Our results suggest that administration of gadobutrol at higher dosage and higher concentration increases sensitivity to perfusion alterations. These results are potentially useful for perfusion-weighted imaging of the human brain, because the volume of contrast agent will be reduced if a solution with higher concentration is used. When using contrast agents in higher concentrations for human examinations, a significant signal decrease may be achieved also with the low doses (.1-.15 mmol/kg). PMID- 9400857 TI - Discrimination of metabolite from lipid and macromolecule resonances in cerebral infarction in humans using short echo proton spectroscopy. AB - Short-echo proton spectroscopy allows the noninvasive study of metabolites, lipids, and macromolecules in stroke patients, but spectra are difficult to interpret and quantify because narrow metabolite peaks are added to a broad background of lipid and macromolecule peaks. "Metabolite nulling" was used to distinguish the lactate peak from underlying lipid and macromolecule peaks. Increases in the lipid and macromolecule peaks were initially observed within the region of infarction in all patients, and further increases in lipid peaks were seen in five of the six patients during the following 6 weeks. The initial high lactate concentration decreases during the first 2 weeks after stroke, whereas lipid and macromolecule signals show a persistent elevation during the same period. Differences in the time courses of the observed changes suggest that lipid, macromolecule, and lactate signals arise from more than one source. PMID- 9400858 TI - Centric ordering is superior to gradient moment nulling for motion artifact reduction in EPI. AB - Echo-planar imaging (EPI) is sensitive to motion despite its rapid data acquisition rate. Compared with traditional imaging techniques, it is more sensitive to motion or flow in the phase-encode direction, which can cause image artifacts such as ghosting, misregistration, and loss of spatial resolution. Consequently, EPI of dynamic structures (eg, the cardiovascular system) could benefit from methods that eliminate these artifacts. In this paper, two methods of artifact reduction for motion in the phase-encode direction are evaluated. First, the k-space trajectory is evaluated by comparing centric with top-down ordered sequences. Next, velocity gradient moment nulling (GMN) of the phase encode direction is evaluated for each trajectory. Computer simulations and experiments in flow phantoms and rabbits in vivo show that uncompensated centric ordering produces the highest image quality. This is probably due to a shorter readout duration, which reduces T2* relaxation losses and off-resonance effects, and to the linear geometry of phantoms and vessels, which can obscure centric blurring artifacts. PMID- 9400859 TI - The effects of incomplete breath-holding on 3D MR image quality. AB - The purpose of this study was to investigate how fast three-dimensional (3D) MR image quality is affected by breath-holding and to develop an optimal breath holding strategy that minimizes artifact in the event of an incomplete breath hold. A computer model was developed to study variable-duration breath-holds during fast 3D imaging. Modeling was validated by 3D gradient-echo imaging performed on 10 volunteers. Signal-to-noise ratio (SNR) and image blur were measured for both simulated and clinical images. Insights gained were applied to clinical 3D gadolinium-enhanced MR angiography. Breath-holding significantly improved abdominal 3D MR image quality. Most of this benefit could be achieved with a breath-hold fraction of 50% if it occurred during acquisition of central k space. Breath-holding during peripheral k-space acquisition, however, had no significant benefit. Respiratory motion artifact on fast 3D MRI occurring when a patient fails to suspend respiration for the entire scan duration can be minimized by collecting central k space first (centric acquisition) so that premature breathing affects only the acquisition of peripheral k space. PMID- 9400860 TI - Uptake of dextran-coated monocrystalline iron oxides in tumor cells and macrophages. AB - Although several dextran-coated iron oxide preparations are in preclinical and clinical use, little is known about the mechanism of uptake into cells. As these particles have been shown to accumulate in macrophages and tumor cells, we performed cellular uptake and inhibition studies with a prototypical monocrystalline iron oxide nanoparticle (MION). MION particles were labeled with fluorescein isothiocyanate or radioiodinated and purified by gel permeation chromatography. Two preparations of MION particles were used in cell experiments: nontreated MION and plasma-opsonized MION purified by gradient density purification. As determined by immunoblotting, opsonization resulted in C3, vitronectin, and fibronectin association with MION. Incubation of cells with fluorescent MION showed active uptake of particles in macrophages both before and after opsonization. In C6 tumor cells, however, intracellular MION was only detectable in dividing cells. Quantitatively, 125I-labeled MION was internalized into cells with uptake values ranging from 17 ng (in 9L gliosarcoma) to 970 ng iron per million cells for peritoneal macrophages. Opsonization increased MION uptake into macrophages sixfold, whereas it increased the uptake in C6 tumor cells only twofold. Results from uptake inhibition assay suggest that cellular uptake of nonopsonized (dextran-coated) MION particles is mediated by fluid-phase endocytosis, whereas receptor-mediated endocytosis is presumably responsible for the uptake of opsonized (protein-coated) particles. PMID- 9400862 TI - Detection of primary breast cancer in women with known adenocarcinoma metastatic to the axilla: use of MRI after negative clinical and mammographic examination. AB - The purpose of this study was to evaluate the ability of MRI to identify a primary site of malignancy in the breast of patients who present clinically with ipsilateral lymph nodes containing metastatic carcinoma but whose physical and mammographic examination are negative. MRI of the breast was performed on four patients using a variety of imaging parameters, all with and without gadolinium contrast. All patients had biopsy-proven adenocarcinoma of the ipsilateral axilla, with negative physical and mammographic examinations. Foci of enhancement assessed visually on precontrast and postcontrast scans (n = 1) and on subtraction studies (n = 3) were considered suspicious under the clinical circumstances defined for this study. Lesions identified on MRI were re identified on ultrasound examination and either preoperative localization for excisional biopsy or tissue sampling was performed. Surgery was performed and histopathologic correlation was obtained in all cases. Primary sites of breast carcinoma were identified in all four patients, with multiple sites of malignancy identified in three of four patients. Breast conservation therapy was made possible for three of four patients based on the results of the MRI study showing sites of malignancy and no features of cancer elsewhere in the breast. Follow-up data of 1, 2, and 5 years of these patients show no evidence of recurrent disease. MRI of the breast is a useful technique for identifying primary sites of malignancy in patients presenting with ipsilateral lymph nodes positive for metastatic adenocarcinoma when the physical and mammographic examinations are negative. PMID- 9400861 TI - Volume MRI and MRSI techniques for the quantitation of treatment response in brain tumors: presentation of a detailed case study. AB - Patients with primary brain tumors may be considered for several different treatments during the course of their disease. Assessments of disease progression and response to therapy are typically performed by visual interpretation of serial MRI examinations. Although such examinations provide useful morphologic information, they are unable to reliably distinguish active tumor from radiation necrosis. This poses a particular problem in the assessment of response to localized radiation therapies such as gamma knife radiosurgery. In this paper, we present methodology for evaluating changes in tissue morphology and metabolism based on serial volumetric MRI and magnetic resonance spectroscopic imaging (MRSI) examinations. Registration and quantitative analysis of these data provide measurements of the temporal and spatial distributions of gadolinium enhancement and of N-acetylasparate, choline, creatine, and lactate/lipid. The key features of this approach and the potential clinical benefits are illustrated by a detailed analysis of six serial MRI/MRSI examinations and three serial 1-[F-18] fluoro-2-deoxy-D-glucose (FDG) positron emission tomography (PET) studies on a patient with a recurrent anaplastic astrocytoma. PMID- 9400863 TI - First pass MRA of the abdomen: ultrafast, non-breath-hold time-of-flight imaging using Gd-DTPA bolus. AB - The authors describe a new fast imaging sequence that can produce projection angiograms of the abdominal vessels at a rate of 2 to 3 frames per second. The result is a versatile imaging technique that can track the arrival of a bolus of contrast in major vessels. With very fast data acquisition, gross patient motion is not a problem, and routine vascular projection studies may be performed without the need for breath-holding. This method is compatible with later high resolution three-dimensional gradient echo studies using contrast agents and may, in fact, be used as an accurate timing protocol to gauge the arrival time of contrast in various segments of the abdominal vessels. Compared with echo planar imaging, this method has the advantages of avoiding susceptibility artifacts and depicting retroperitoneum and other abdominal fat-containing landmarks and does not require extensive hardware modifications for a clinical system. PMID- 9400864 TI - Breath-hold R2* mapping with a multiple gradient-recalled echo sequence: application to the evaluation of intrarenal oxygenation. AB - Blood oxygenation level dependent (BOLD) MRI is sensitive to changes in regional oxygen supply versus demand and is therefore potentially useful in evaluating susceptibility to ischemic injury. Recently, we have demonstrated the use of BOLD MRI to evaluate intrarenal oxygenation using single shot echo-planar imaging (EPI). Here, we present an alternate implementation of BOLD MRI sequence, using multiple gradient echoes, that does not require any specialized hardware. PMID- 9400865 TI - Elimination of Nyquist ghosts in MRI by using fast linogram imaging. AB - Timing inaccuracies between the even and odd echoes lead to the formation of Nyquist ghosts in conventional (blipped) echo-planar imaging (EPI). A fast radial scanning method based on the linogram sampling geometry was designed and implemented. No ghosting effects are seen with this scheme, and correction for timing inaccuracies is performed using simple postprocessing steps before reconstruction. PMID- 9400867 TI - Aboriginal reconciliation: a role for psychiatrists? AB - Australian Aboriginal people are marginalised, severely disadvantaged and subject to discrimination. The Report of the National Inquiry into the Separation of Aboriginal and Torres Strait Islander Children from their Families has focused international attention on their plight. However, the Council for Aboriginal Reconciliation and the High Court Mabo decision provide avenues for change. This paper asks: is there a role for psychiatrists in the reconciliation process? PMID- 9400866 TI - The effect of mechanical deformation on magnetic properties and MRI artifacts of type 304 and type 316L stainless steel. AB - The purpose of this study was to evaluate the influence of composition and deformation of biomedical stainless steels on mechanical properties, magnetic properties, and MRI artifacts. Type 304 and Type 316L samples were prepared using standard wire-drawing techniques. Mechanical properties were determined using standard test methods. The amount of ferromagnetic phase present was estimated using a Severn Gage and x-ray diffraction. Magnetic field attraction and artifacts were determined using previously described techniques. The strength of both steels increased significantly with increasing deformation. None of the type 316L wires transformed to the magnetic phase. The amount of magnetic phase in the type 304 wires increased with increasing deformation. There was no magnetic field attraction, and artifacts were minimal for all of type 316L wires and the undeformed type 304 wire. Deflection and artifacts were significant for the deformed type 304 stainless steel. These results provide guidance regarding the use of type 304 and type 316L stainless steels for bioimplants. In this regard, type 316L stainless steel seems to be a more acceptable material with respect to MR compatibility. PMID- 9400868 TI - The NHMRC Psychiatric Epidemiology Research Centre. National Health and Medical Research Council. PMID- 9400869 TI - Training in psychiatry: an examination of trainee perceptions. Part 1. AB - OBJECTIVE: To look at the perceptions of New South Wales (NSW) psychiatric trainees in relation to their training experiences and the role and quality of the consultant-registrar relationship. METHODS: A self-report questionnaire was developed to probe trainee perceptions of the consultant-trainee relationship in all those who had completed at least 1 year of training in psychiatry (n = 138) in NSW, as well as all consultants who had completed their training in the last 5 years (n = 95). Test-retest reliability was assessed at 3 months for each of the subscales (r = 0.70-0.89) and found to be acceptable. Validity issues are discussed. RESULTS: The results are discussed with special reference to the perceived competence, availability, breadth of knowledge and willingness to accept responsibilities of the supervising consultant. Consultant competence as a clinician was consistently rated as more important than being emotionally supportive. CONCLUSION: In addressing these issues, we aim to increase the degree of self-consciousness and reflectiveness of the profession of psychiatry within the Australian context. If there is to be a substantial shift for the better in trainees' perceptions of consultants, it is likely that the general consultant experience will have to be improved rather than providing small amounts of exposure to high quality consultants. PMID- 9400870 TI - Adverse experiences in psychiatric training. Part 2. AB - OBJECTIVE: To examine the perceptions of New South Wales (NSW) psychiatric trainees in relation to their training, the adverse events they experienced and the role and quality of the consultant-registrar relationship. METHOD: A self report questionnaire was developed to probe trainee perceptions of the consultant trainee relationship and adverse events during training in all those who had completed at least 1 year of training in psychiatry (n = 138) in NSW, as well as all consultants who had completed their training in the last 5 years (n = 95). All subjects were asked to rate the frequency and relative impact of 20 adverse experiences with the opportunity to proffer adversities not listed. They were also asked to rate their experience of their consultants in relation to the adversity. RESULTS: The results from The Training Impact Study exploring adverse events experienced by NSW trainees are presented. Assault by a patient and suicide of a patient are identified as the most stressfull adversities of training in psychiatry. However, more general concerns such as educational and emotional neglect by supervisors, observing consultant maltreatment of patients, exam failure and conflict between consultants were also identified and discussed. CONCLUSIONS: The high response rate of both trainees and consultants gives these results a level of representative validity. Recommendations in relation to future training and research are put forward. Specific training in the management of potentially assaultive patients and facilitating trainee recovery from assault or threat of assault should be a priority of the Royal Australian and New Zealand College of Psychiatrists. Support and education in relation to patient suicide is also important. Training and recognition of teachers within the College should be encouraged. PMID- 9400871 TI - Peer review groups: problems and solutions. AB - OBJECTIVE: To elucidate and delineate principles for dealing with problems encountered in peer review groups. METHOD: A workshop format was utilised in which a number of interested psychiatrists were invited to role-play specific problems which occurred in established peer review groups, and reported in recent research studies. Process notes or videotapes were used to record proceedings. Principles were delineated for the management of five specific problems via role play and discussion. RESULTS: Principles have been delineated for the management of the following problems reported by peer review groups: dominating group member; member with disturbing behaviour; member providing inadequate treatment; member transgressing treatment boundaries; and group members strongly disagreeing about the management of a case. CONCLUSION: Problems encountered within peer review groups were adequately replicated during role-played enactments. The workshop format offered a means of engaging with these, allowing exploration and subsequent resolution of problems which had not proved resolvable in the real group peer review situation. PMID- 9400873 TI - Experience, knowledge and attitudes of child psychiatrists regarding electroconvulsive therapy in the young. AB - OBJECTIVE: To ascertain the experience, knowledge and attitudes of Australian and New Zealand child psychiatrists in relation to electroconvulsive therapy (ECT) in the young in order to determine whether they would be willing and able to provide an opinion if consulted about children or adolescents in whom ECT is proposed. METHOD: A 28-item questionnaire was posted to all members of the Faculty of Child and Adolescent Psychiatry living in Australia or New Zealand. RESULTS: Eighty three percent (n = 206) answered the questionnaire. Forty percent rated their knowledge about ECT in the young as nil or negligible. Having had patients treated with ECT was the best predictor of possessing some knowledge. Thirty-nine percent believed that ECT was unsafe in children compared to 17% for adolescents and 3% for adults. Almost all (92%) respondents believed child psychiatrists should be consulted in all cases of persons under 19 in whom ECT was recommended. The vast majority believed the Faculty or College should have guidelines relating to ECT use in this group and that it would be useful to have a national register of young persons treated with ECT. CONCLUSIONS: Child and adolescent psychiatrists wish to be involved in the process of ECT treatment in young people. At the same time, there are gaps in their knowledge. This will need to be remedied, particularly if formal guidelines advocating their involvement are introduced. PMID- 9400872 TI - The relationship between low family income and psychological disturbance in young children: an Australian longitudinal study. AB - OBJECTIVE: This study examines the relationship between low family income (LFI) experienced at different points in time, chronic low income status and its impact on child behaviour measured at 5 years of age. METHOD: Longitudinal data from the Mater University Study of Pregnancy were used to measure LFI in families at three points in time (the antenatal period, 6 months post birth and at 5 years cf age). Outcome variables were three independent groups of behaviour problems labelled as externalising, social, attentional and thought (SAT) problems, and internalising problems. These groups were developed from the Child Behaviour Checklist. An analysis based on logistic regression modelling was carried out examining the relationship between LFI and a range of intermediate variables known to be associated with child behaviour problems. RESULTS: The more often families experienced low income, the higher the rate of child behaviour problems at age 5. Low family income was still independently associated with SAT behaviour problems after controlling for smoking in the first trimester, parenting styles, maternal depression and marital dysharmony at age 5. The association between LFI and internalising and externalising behaviour problems was largely mediated by maternal depression. CONCLUSION: Low family income is a significant factor in the aetiology of a variety of child behaviour problems. The mechanisms involved in the link between LFI and childhood internalising and externalising behaviours involve the exposure of the children to maternal depression. However, the relationship between LFI and SAT behaviour problems remains to be elucidated. PMID- 9400874 TI - A profile of children and adolescents in a psychiatric unit: multidomain impairment and research implications. AB - OBJECTIVE: The scientific literature has not kept pace with the evolution of child and adolescent psychiatric inpatient units, including their nature, patient profile, philosophical orientation and efficacy. This study aims to establish a comprehensive, multimodal description of the population served by an inpatient psychiatric treatment facility for children and adolescents. METHOD: A multidisciplinary assessment regime including psychiatric, medical, speech and language examination, observer rating and patient self-report of psychopathology was used to assess 58 consecutive patients over a 20-month period. RESULTS: In addition to a prevalence of disruptive behaviour disorders of 67% and a high rate of comorbidity with other psychiatric conditions, a breadth of impairment was demonstrated in many areas. Significantly decreased measures of socialisation, communication, daily living skills, self-esteem, intelligence and physical health are reported. Moderate to severe language handicap was found in 40% of patients. CONCLUSIONS: The inpatient population of children and adolescents exhibited not only a high rate of disruptive behaviour disorders, frequently comorbid with other psychiatric conditions, but also high levels of physical, speech, language and living skills impairment. This finding supports the need for multimodal, multidisciplinary evaluation and treatment in this population. Outcome research evaluating treatment effectiveness must also account for the wide-ranging disabilities of these children and adolescents. PMID- 9400875 TI - Paradox, persecution and the double game: psychotherapy in anorexia nervosa. AB - OBJECTIVE: In anorexia nervosa, the interface between psychotherapy and effective physical management continues to arouse dissent. For this reason, theoretical underpinnings and their practical applications were explored. METHOD: A selective review of the literature was based on the authors' clinical experience in hospital treatment settings. RESULTS: The psychosomatic paradigm, the threat to life, the reward characteristics and the developmental trajectory of anorexia nervosa necessitate a particular form of integrated psychotherapy. CONCLUSIONS: The approach is essentially that of self psychology but with an important departure which must be addressed in the therapeutic process. This is exemplifed in the paradox of persecution by refeeding and the curious double game that evolves. PMID- 9400876 TI - The comorbidity of anxiety and depression. AB - OBJECTIVE: This study explored the effect of comorbid anxiety disorders in patients admitted to an inpatient specialist Mood Disorders Unit for the treatment of a primary major depressive episode. METHOD: Subjects were assessed on admission and discharge. DSM-III-R diagnoses for major depression and anxiety disorders were established using CIDI-Auto; comorbid anxiety disorders were coexistent in time with the major depression, with both conditions meeting diagnostic criteria at the time of assessment. Severity of illness was assessed using the Hamilton Depression/Melancholia Scale, the revised Hamilton Anxiety Scale and the revised Beck Depression Inventory. RESULTS: For the analysis, the study cohort was divided into three groups: depression alone (n = 33), one comorbid anxiety disorder (n = 15), and two or more comorbid anxiety disorders (n = 24). No particular anxiety disorder predominated. Interestingly, the presence or absence of comorbid anxiety with severe major depression made no significant difference to treatment choice or outcome results. Specifically, there was no significant difference between the three groups in the utilisation of electroconvulsive therapy and pharmacotherapy (including antidepressants, benzodiazepines and neuroleptics); all subjects improved significantly on both depression and anxiety ratings, and length of inpatient stay did not vary significantly between the three groups. CONCLUSIONS: The existence of comorbid anxiety disorders in those patients who presented for treatment of a primary major depressive episode did not significantly effect choice of treatment or treatment outcome, suggesting that there is a close interrelationship between the two conditions. PMID- 9400877 TI - The molecular genetics of schizophrenia: an update. AB - OBJECTIVE: This paper aims to summarise the latest molecular genetic findings in schizophrenia, while providing background information on a number of relevant methodological issues. METHOD: Accumulative genetic data indicate that schizophrenia is a genetically complex disease with an unclear mode of transmission. The development and rapid progression of molecular genetics have provided a wide variety of methods to search for genes predisposing to human disease. The genetic basis for a number of the simpler diseases has been identified and characterised using these methods. More recently, progress has been made in identifying genes predisposing to the genetically more complex diseases such as diabetes mellitus, multiple sclerosis, bipolar disorder and schizophrenia. RESULTS: The latest findings on chromosomes 3, 6, 8, 13, 18 and 22 and on the X chromosome are reviewed. CONCLUSIONS: There is now suggestive support for three susceptibility loci (6p24-22, 8p22-21 and 22q12-q13.1) for schizophrenia, and it is likely that other regions will emerge from studies now in progress. Finding and then characterising genes within these loci will require long-term commitment and systematic efforts in clinical, laboratory and analytical fields. PMID- 9400879 TI - Adverse psychological impact of operative obstetric interventions: a prospective longitudinal study. AB - OBJECTIVE: This paper reports the findings of a prospective longitudinal study of 272 nulliparous pregnant women, which investigated as one of its objectives the psychological sequelae of obstetric procedures. METHOD: Participants completed structured interviews and standardised, published psychometric questionnaires, including the Rosenberg Self-Esteem Scale and the Profile of Mood States late in pregnancy and again early in the postpartum period. RESULTS: Little evidence was found to support the notion that the total number of obstetric interventions was linked to a deterioration in postpartum mood. Significant adverse psychological effects were associated with the mode of delivery. Those women who had spontaneous vaginal deliveries were most likely to experience a marked improvement in mood and an elevation in self-esteem across the late pregnancy to early postpartum interval. In contrast, women who had Caesarean deliveries were significantly more likely to experience a deterioration in mood and a diminution in self-esteem. The group who experienced instrumental intervention in vaginal deliveries fell midway between the other two groups, reporting neither an improvement nor a deterioration in mood and self-esteem. CONCLUSIONS: The findings of this study suggest that operative intervention in first childbirth carries significant psychological risks rendering those who experience these procedures vulnerable to a grief reaction or to posttraumatic distress and depression. PMID- 9400878 TI - An evaluation of the effectiveness of a consultation-liaison psychiatry service in general practice. AB - OBJECTIVE: This study evaluated the 6-month outcome of patients referred by their general practitioner (GP) to a consultation-liaison (C-L) psychiatry service provided to eight group general practices. METHOD: Over a 12-month period, there were 307 referrals to the C-L psychiatry service of whom 86 consented to take part in an outcome study. Two different control groups were examined comprising patients seen by the same GPs but not referred to the C-L service, who were matched with the C-L referrals on the basis of either demographic characteristics (n = 86) or initial symptomatology (n = 59). Clinical interviews were conducted at recruitment to the outcome study using the Composite International Diagnostic Interview (CIDI), while postal questionnaires were used at both the initial and 6 month assessments. RESULTS: Data reported include DSM-III-R clinical audit and CIDI diagnoses, changes in current symptomatology (SCL-90-R) and changes in global ratings of physical health, emotional health, social relationships and ability to perform everyday duties. Consultation-liaison referrals without symptom-matched controls (n = 27), being patients with higher levels of symptoms initially, were more likely to be referred to other psychiatric services for treatment. They also showed more marked improvement over time on the selected outcome measures. However, there were no significant differences in the patterns of change over time between symptom-matched C-L referrals and their non-referred controls. CONCLUSIONS: The findings from the 6-month outcome study raise doubts about the overall benefit of the current C-L service relative to usual GP care. Improving the quality of psychiatric care in general practice is likely to require a range of interrelated strategies, including C-L psychiatry services, GP education and well-functioning links with public mental health services. PMID- 9400880 TI - Using action research to facilitate organisational change in mental health service delivery. AB - OBJECTIVE: To report how well a particular mental health service was responding to National and State mental health policies by emphasising community-based clinical care for patients. METHOD: The method involved analysis and action in response to information obtained from a consecutive sample of 100 people admitted to a psychiatric hospital ward from a geographical catchment area. RESULTS: It was found that community mental health staff were working without adequate support and were not able to effectively implement early intervention and relapse prevention strategies. Concern about these findings led to organisational change using action research methodology and some of these changes are discussed. CONCLUSIONS: Such methodology was a useful tool to identify and achieve organisational change in psychiatric services. PMID- 9400882 TI - The case-conferencing project: a first step towards shared care between general practitioners and a mental health service. AB - OBJECTIVES: The objectives of this study were: (i) to improve general practitioners' knowledge of the mental disorders they commonly treat, and to increase their confidence in managing people with these disorders; and (ii) to increase general practitioners' familiarity with the Logan-Beaudesert Mental Health Service. METHOD: Eleven general practitioners met with psychiatrists from the Logan-Beaudesert Mental Health Service in six 2-hour sessions held at monthly intervals. Each session comprised a teaching component, a consumer interview and a case-conference. Outcomes were measured using an objective test of general practitioners' knowledge, a subjective test of their confidence in dealing with mental health problems, and satisfaction surveys for participating consumers, general practitioners and psychiatrists. RESULTS: On the objective test, the scores of 10 out of the 11 general practitioners improved (p < 0.05). On the subjective test, the ranked scores improved in nine out of the 11 cases (p < 0.05). Consumers, general practitioners and psychiatrists expressed their satisfaction with the format and content of the course. CONCLUSIONS: Having improved the knowledge of a group of general practitioners who are familiar with the functioning of the Logan-Beaudesert Mental Health Service, the stage is now set to proceed to the next step: the shared-care project. PMID- 9400881 TI - Who is a heavy service user? Preliminary development of a screening instrument for prospective consumers of a mobile intensive treatment team. AB - OBJECTIVE: The mobile intensive treatment team (MITT) of the Valley Integrated Adult Mental Health Service in Brisbane, Australia, aims to provide services in the community to people with severe and persistent mental illness who have traditionally been heavily reliant on inpatient services (i.e. heavy service users). The MITT screening instrument (MITTSI) was developed to provide an objective measure to appropriately identify patients for referral to the service. METHOD: A literature review and a panel of multidisciplinary clinicians were consulted to identify a list of specific attributes that would assist in the detection of heavy service users. These attributes were then formulated into an easy-to-administer screening instrument entitled the MITTSI. The MITTSI was administered in an interview format to MITT case managers (intensive case management) and to case managers in standard case management with prospective MITT patients (prospective heavy service users). RESULTS: Analyses of the responses indicated support for the MITTSI as a valid screening instrument in identifying heavy service users and for determining appropriate patients for referral to the MITT. CONCLUSION: The MITTSI is an easy-to-administer screening instrument which provides clear guidelines for inclusion and exclusion, and is an objective measure regarding the patients' urgency for referral to the MITT. Follow-up of the MITTSI within a broader, longer-term project will attempt to further refine the MITTSI and to further determine its validity. Outcomes will be published at a later stage. PMID- 9400883 TI - How are psychotic symptoms perceived? A comparison between patients, relatives and the general public. AB - OBJECTIVE: The aim of this study is to compare patients with schizophrenia with their relatives and the general public in their attitudes towards schizophrenic psychotic symptoms. METHOD: We used a case vignette depicting a person with typical schizophrenic psychotic symptoms and compared the attitudes of 44 inpatients and 47 outpatients with schizophrenia, 48 of their relatives and 43 members of the general public. We also compared the attitudes of patients with schizophrenia to their own symptoms and the symptoms described in the vignette. RESULTS: Subjects from the general public tended not to recognise psychotic symptoms as features of mental illness and tended not to consider drug treatment and hospitalisation as required. Sex, education level as well as previous contact with the mentally ill were found to be significant determinants of attitude. The levels of symptom awareness in patients with schizophrenia and their relatives are higher but still relatively low. In addition, we found that patients with schizophrenia who correctly appraised psychotic symptoms in another person were also aware of their own mental symptoms and need of treatment. CONCLUSIONS: The level of recognition of psychotic symptoms and awareness of a need for treatment are low in the general public, as well as in patients with schizophrenia and their relatives. These findings are discussed in relation to the assessment of insight in patients and a need for psychoeducational programs for each group. PMID- 9400884 TI - Psychosis following acute alteration of thyroid status. AB - OBJECTIVE: Mania with psychotic features presenting following abrupt normalisation of thyroid function from severe Graves's disease is reported. CLINICAL PICTURE: A 33-year-old man with severe, untreated Graves's disease was treated aggressively, with rapid restoration of normal serum thyroid hormone levels. Symptoms of mania and psychosis then developed. TREATMENT: Time limited antipsychotic medication and continuing medical treatment. OUTCOME: There was resolution of psychiatric symptoms. CONCLUSIONS: The association of mania and psychosis with thyroid disease is rare, but aggressive medical treatment and rapid restoration of normal serum thyroid levels may increase the risk of the emergence of such symptoms. PMID- 9400885 TI - Erotomania associated with temporal lobe abnormalities following radiotherapy. AB - OBJECTIVE: The aetiology of primary erotomania is usually discussed in terms of psychological disturbance in the patient. A case associated with demonstrable left temporal lobe abnormalities is described. CLINICAL PICTURE: An elderly female patient presented with the delusion of being loved by a physician who had treated her previously. She had received radiotherapy to her left periorbital area in childhood. Structural and functional neuroimaging revealed medial temporal lobe damage. TREATMENT AND OUTCOME: She was treated with haloperidol with moderate improvement in her distress. CONCLUSIONS: This case illustrates the contribution of both cerebral injury and psychosocial factors in the eventual development of this unusual psychiatric syndrome. PMID- 9400886 TI - Charles Bonnet syndrome with major depression in a Chinese middle-aged man. AB - OBJECTIVE: To describe a middle-aged patient with Charles Bonnet syndrome (CBS) suffering from concurrent major depression. CLINICAL PICTURE: A 41-year-old Chinese man with retinitis pigmentosa developed complex and vivid visual hallucinations. This was followed by the onset of major depressive illness. His visual hallucinations were greatly influenced by his cultural background and changed during the course of his depression. TREATMENT: The patient was treated with imipramine. OUTCOME: The patient had a relapse of depression due to his non compliance. He recovered after the resumption of imipramine but the visual hallucinations persisted. CONCLUSIONS: Patients suffering from CBS can have another concurrent psychiatric illness. The content and form of this patient's visual hallucinations were modified by his cultural background and depressive illness. Sensory deprivation is suggested to be the pathogenic mechanism of his visual hallucination. PMID- 9400887 TI - Obsession with infidelity: another case and some views. PMID- 9400888 TI - Existing brain condition may predispose to SSRI-induced extrapyramidal symptoms. PMID- 9400889 TI - Hired guns. PMID- 9400890 TI - Legalistic abuse of psychiatric diagnoses and our own future. PMID- 9400891 TI - Urinary tract infection and delusion of pregnancy. PMID- 9400892 TI - Introduction: COPD--the role of infection. PMID- 9400893 TI - Defining subsets of patients with chronic bronchitis. PMID- 9400894 TI - Guidelines for the treatment of acute exacerbations of chronic bronchitis. PMID- 9400895 TI - Antibiotic selection and dosing for the treatment of acute exacerbations of COPD. PMID- 9400896 TI - Introduction: surviving septic shock. PMID- 9400897 TI - Proinflammatory and anti-inflammatory cytokines as mediators in the pathogenesis of septic shock. PMID- 9400898 TI - New therapies in sepsis. PMID- 9400899 TI - Current status of clinical trials with anti-TNF. PMID- 9400900 TI - Ethical considerations on the use of monoclonals in the managed care environment. PMID- 9400901 TI - The SYST-EUR Study--calcium channel blockers coming of age? European Trial on Isolated Systolic Hypertension in the Elderly. PMID- 9400902 TI - Pseudo-hypertension in the elderly: still hazy, after all these years. PMID- 9400903 TI - Stroke, blood pressure and antihypertensive therapy. PMID- 9400904 TI - Moxonidine: a review. AB - Moxonidine is an imidazoline compound which acts on I1 imidazoline 'receptors' in the central nervous system to reduce blood pressure. This novel mechanism of action is claimed to lead to fewer adverse effects than the older centrally acting agents such as clonidine. In this review we examine the drug's pharmacology, clinical pharmacokinetics, efficacy as an antihypertensive agent including comparative studies with pre-existing drugs, and adverse effect profile. With a growing number of effective antihypertensive agents already available to the clinician, it is not yet clear whether moxonidine represents a significant advance in hypertension management. PMID- 9400905 TI - Effects of antihypertensive agents on circadian blood pressure in hypertensive patients with previous brain infarction. AB - To evaluate the effects of antihypertensive agents on the circadian blood pressure (BP) of patients with previous brain infarction, the ambulatory BP was measured non-invasively for 24 h before and after administration of antihypertensive agents. One hundred milligrams of acebutolol twice daily (n = 15) is effective in lowering the BP during the daytime, but has little effect during the night and the morning. Twenty milligrams of slow-release nifedipine twice daily (n = 14) produced a consistent reduction in the BP over the entire 24 h period and effectively blunted the rise in BP in the morning. Captopril (12.5 mg) twice daily (n = 15) produced a mild reduction in BP with little change in the circadian pattern. The slow-release nifedipine group had the greatest decrease in mean systolic and diastolic BP. The heart rate significantly increased after administration of slow-release nifedipine and decreased after administration of acebutolol. To reduce stroke recurrence, we should consider the effects of antihypertensive agents on circadian BP in hypertensive patients with previous brain infarction. PMID- 9400906 TI - The effectiveness of exercise training in lowering blood pressure: a meta analysis of randomised controlled trials of 4 weeks or longer. AB - OBJECTIVE: To identify the features of an optimal exercise programme in terms of type of exercise, intensity and frequency that would maximise the training induced decrease in blood pressure (BP). DATA IDENTIFICATION: Trials were identified by a systematic search of Medline, Embase and Science Citation Index (SCI), previous review articles and the references of relevant trials, from 1980 until 1996, including only English language studies. STUDY SELECTION: The inclusion criteria were limited to randomised controlled trials of aerobic or resistance exercise training conducted over a minimum of 4 weeks where systolic and diastolic BP was measured. RESULTS: A total of 29 studies (1533 hypertensive and normotensive participants) were included, 26 used aerobic exercise training, two trials used resistance training and one study had both resistance and aerobic training groups. Aerobic exercise training reduced systolic BP by 4.7 mm Hg (95% CI: 4.4, 5.0) and diastolic BP by 3.1 mm Hg (95% CI: 3.0, 3.3) as compared to a non-exercising control group, however, significant heterogeneity was observed between trials in the analysis. The BP reduction seen with aerobic exercise training was independent of the intensity of exercise and the number of exercise sessions per week. The evidence for the effect of resistance exercise training was inconclusive. CONCLUSIONS: Aerobic exercise training had a small but clinically significant effect in reducing systolic and diastolic BP. Increasing exercise intensity above 70% VO2 max or increasing exercise frequency to more than three sessions per week did not have any additional impact on reducing BP. PMID- 9400907 TI - Efficacy and position of endurance training as a non-drug therapy in the treatment of arterial hypertension. AB - Regular conditioning has been well documented to exert a beneficial effect on cardiovascular risk factors and to improve overall cardiovascular health and to reduce the incidence of coronary disease. There are conflicting results concerning the effect of physical exercise on blood pressure (BP) in hypertensive patients and its importance in the treatment of hypertension. Therefore 10 male patients with mild arterial hypertension were studied in order to define the BP response to long-term aerobic training (60 min twice a week) under resting conditions, during standardised ergometric workload, during isometric exercise, during cold pressor testing and during 24-h BP monitoring. After 18 months of regular training there were significant reductions in arterial pressures at rest, during and after standardised ergometry and during isometric and cold pressor testing when compared with pre-training. The heart rate also decreased significantly during exercise testing thus implying a decrease in myocardial oxygen consumption. After long-term training, a reduction in systolic and diastolic BP could also be shown during 24-h ambulatory BP monitoring. These results demonstrate that long-term aerobic training leads to a decrease in systolic and diastolic BP at rest, during exercise and during 24-h BP monitoring and imply a beneficial effect in the management of hypertension that is nearly comparable to that of drug therapy. PMID- 9400908 TI - Avoiding the supine position during sleep lowers 24 h blood pressure in obstructive sleep apnea (OSA) patients. AB - Obstructive sleep apnea (OSA), is a common clinical condition affecting at least 2-4% of the adult population. Hypertension is found in about half of all OSA patients, and about one-third of all patients with essential hypertension have OSA. There is growing evidence that successful treatment of OSA can reduce systemic blood pressure (BP). Body position appears to have an important influence on the incidence and severity of these sleep-related breathing disturbances. We have investigated the effect of avoiding the supine position during sleep for a 1 month period on systemic BP in 13 OSA patients (six hypertensives and seven normotensives) who by polysomnography (PSG) were found to have their sleep-related breathing disturbances mainly in the supine position. BP monitoring was performed by 24-h ambulatory BP measurements before and after a 1 month intervention period. We used a simple, inexpensive method for avoiding the supine posture during sleep, namely the tennis ball technique. Of the 13 patients, all had a reduction in 24-h mean BP (MBP). The mean 24-h systolic/diastolic (SBP/DBP) fell by 6.4/2.9 mm Hg, the mean awake SBP/DBP fell by 6.6/3.3 mm Hg and the mean sleeping SBP/DBP fell by 6.5/2.7 mm Hg, respectively. All these reductions were significant (at least P < 0.05) except for the sleeping DBP. The magnitude of the fall in SBP was significantly greater in the hypertensive than in the normotensive group for the 24 h period and for the awake hours. In addition, a significant reduction in BP variability and load were found. Since the majority of OSA patients have supine-related breathing abnormalities, and since about a third of all hypertensive patients have OSA, avoiding the supine position during sleep, if confirmed by future studies, could become a new non-pharmacological form of treatment for many hypertensive patients. PMID- 9400909 TI - Autonomic nervous system activity in essential hypertension: a comparison between dippers and non-dippers. AB - The present study was undertaken to investigate the changes in autonomic nervous system activity in essential hypertension. Fourteen normotensive controls and 33 age-matched untreated hypertensive subjects, diagnosed by ambulatory blood pressure (ABP) measurement (24-h systolic ABP value over 140 mm Hg or 24-h diastolic ABP over 90 mm Hg, or both) were recruited. ABP and 24-h electrocardiogram were monitored simultaneously. Power spectral analysis of the R R interval was performed by a fast Fourier transformation method and the powers of low frequency (LF; 0.04 to 0.15 Hz) and high frequency (HF; 0.15 to 0.4 Hz) components were obtained. Hypertensive subjects were divided into 'dippers', whose night-time systolic ABP fell by more than 10% of their daytime ABP, and 'non-dippers' in whom this phenomenon was absent. In hypertensive subjects, electrocardiogram monitoring and power spectral analysis were also performed for 5 min before and during 90 degrees tilt. There were no significant differences in the 24-h mean LF/HF power ratio, LF power or HF power between normotensive and hypertensive subjects. A significant negative correlation between the night-time systolic ABP level and the 24-h LF/HF power ratio was found (r= -0.36, P < 0.05) in the hypertensive subjects. A significant positive correlation was found between the 24-h LF/HF power ratio and the percentage nocturnal reduction of the daytime systolic ABP in hypertensive subjects (r = +0.40, P < 0.01). The 24-h LF/HF power ratio was significantly lower in non-dippers than in dippers (2.09 +/ 1.06 vs 3.24 +/- 0.97, P < 0.01). The mean daytime LF/HF power ratio was significantly lower in non-dippers than in dippers (2.50 +/- 1.43 vs 4.08 +/- 1.27, P < 0.01). The night-time LF/HF power ratio was not significantly different between the two groups. The LF/HF power ratio increased significantly in dippers (from 1.32 +/- 1.95 to 4.65 +/- 1.54, P < 0.001) during 90 degrees tilt, but there was no significant change in the LF/HF power ratio in non-dippers during tilt (from 1.13 +/- 0.28 to 1.36 +/- 0.78, NS). The 24-h LF/HF power ratio decreased according as the night-time systolic BP elevated in hypertensive subjects. During ambulatory monitoring, the non-dippers showed a significantly lower LF/HF power ratio than the dippers. The LF/HF power ratio increased significantly in dippers, but not in non-dippers during tilting. These results suggest that impaired cardiovascular reflexes might contribute to the decreased sympathovagal balance in non-dipper type hypertension. PMID- 9400910 TI - For how many days should blood pressure be measured at home in older patients before steady levels are obtained? AB - This study investigated the period of time that blood pressure (BP) should be measured at home in older patients in order to obtain steady BP values. Thirty six men and 38 women (> or =60 years) were recruited at one family practice. At one office visit the family physician measured supine, sitting and standing BPs three times consecutively in each position. During 10 consecutive days, BP was measured at home five times daily. The supine and standing BPs were measured once in the morning and in the evening and the sitting BP once at noon. These home BP values were averaged over the first day (1-day), over the first 3 days (3-day) and all 10 days (10-day) of measurements. In both the supine (-5.1 mm Hg) and sitting (-3.8 mm Hg) positions the 10-day average systolic home BP was significantly lower than the corresponding office BP. The opposite was observed for the 10-day average standing home BP values (+7.3/+3.4 mm Hg). Comparison of the 3-day and 10-day average home BP values showed only a significantly lower 10 day than 3-day systolic BP level in the supine position (-1.1 mm Hg, 95% CI -1.9 to -0.2 mm Hg). Repeated measures ANOVA, showed a small but significant decrease over time only for the supine systolic home BP (-0.29 mm Hg per day, 95% CI -0.49 to -0.08 mm Hg per day). We conclude that in older subjects, 3 days of home measurements may suffice to obtain steady values for the sitting and standing BPs. A longer interval might be required for the supine BP. PMID- 9400911 TI - Epidemiological study of hypertension and its determinants in an urban population of North India. AB - OBJECTIVES: To determine age-specific prevalence of hypertension and blood pressure (BP) levels in relation to diet and lifestyle factors in North Indians. DESIGN AND SETTING: Cross-sectional survey in 20 randomly selected streets in Moradabad, North India. SUBJECTS AND METHODS: A total of 1806 subjects from North India (904 males and 902 females) age range 25-64 years. The survey methods were as follows: dietary diaries for 7 days food intake record; BP measurements; physician administered questionnaire and anthropometric measurements. Diagnosis of hypertension was based on new World Health Organization/International Society of Hypertension (WHO/ISH) criteria. Risk factors were assessed based on WHO guidelines. RESULTS: The prevalence of hypertension according to WHO/ISH criteria was 23.7% and by old WHO criteria 13.3%. In the WHO/ISH hypertensive group, isolated diastolic hypertension was present in 47.3% males and 40.6% females. Males have a slightly higher prevalence than females in the young age group, however, the prevalence rates are comparable in the older age groups. In both sexes, the prevalence rates and BP level increased with older age. Multivariate analysis revealed that age, higher body mass index, central obesity and higher socioeconomic status were independently and strongly associated with hypertension in both sexes. Higher dietary fat and salt intake and lower physical activity were weakly but significantly associated with hypertension. CONCLUSION: Association of higher socioeconmic status, higher body mass index and central obesity in North Indian adults with higher fat intake, lower physical activity and higher prevalence and level of hypertension indicate that these populations may benefit by decreasing the dietary fat intake and increasing physical activity, with an aim to decrease central obesity for decreasing hypertension in North Indians. PMID- 9400912 TI - Fibromuscular dysplasia. PMID- 9400913 TI - Intensified insulin therapy--is there anything left to argue? PMID- 9400914 TI - Update of tacrolimus in pancreas transplantation. AB - Pancreas transplantation for the better treatment of diabetes mellitus is becoming an important part of the service offered to diabetic patients requiring renal transplantation. Improvements in surgical technique make this a useful option. A major problem, limiting more extensive use of pancreas transplantation to other diabetic patients, remains the inadequacies of present immunosuppressive regimens. A relatively new agent, FK506 or tacrolimus, is being used increasingly because of perceived benefits over older therapeutic agents. There are concerns about the diabetogenic effect of tacrolimus. These may be dose-related, and low dose tacrolimus regimens, by allowing reduction in dosage of other diabetogenic immunosuppressive agents, have produced encouraging results in pancreas transplantation in many centres. Further improvements in immunosuppressive regimens may widen the clinical implications for pancreas transplantation but identifying the patient group who will most benefit remains a priority. PMID- 9400915 TI - Risk of adverse effects of intensified treatment in insulin-dependent diabetes mellitus: a meta-analysis. AB - While the benefits of intensified insulin treatment in insulin-dependent (Type 1) diabetes mellitus (IDDM) are well recognized, the risks have not been comprehensively characterized. We examined the risk of severe hypoglycaemia, ketoacidosis, and death in a meta-analysis of randomized controlled trials. The MEDLINE database, reference lists, and specialist journals were searched electronically or by hand to identify relevant studies with at least 6 months of follow-up and the monitoring of glycaemia by glycosylated haemoglobin measurements. Logistic regression was used for calculation of combined odds ratios and 95% confidence intervals (95% CI). The influence of covariates was examined by including covariate-by-treatment interaction terms. Methodological study quality was assessed and sensitivity analyses were performed. Fourteen trials were identified. These contributed 16 comparisons with 1028 patients allocated to intensified and 1039 allocated to conventional treatment. A total of 846 patients suffered at least one episode of severe hypoglycaemia, 175 patients experienced ketoacidosis and 26 patients died. The combined odds ratio (95% CI) for hypoglycaemia was 2.99 (2.45-3.64), for ketoacidosis 1.74 (1.27-2.38) and for death from all causes 1.40 (0.65-3.01). The risk of severe hypoglycaemia was determined by the degree of normalization of glycaemia achieved (p=0.005 for interaction term), with the results from the Diabetes Control and Complications Trial (DCCT) in line with the other trials. Ketoacidosis risk depended on the type of intensified treatment used. Odds ratios (95% CI) were 7.20 (2.95-17.58) for exclusive use of pumps, 1.13 (0.15-8.35) for multiple daily injections and 1.28 (0.90-1.83) for trials offering a choice between the two (p = 0.004 for interaction). Mortality was significantly (p = 0.007) increased for causes potentially associated with acute complications (7 vs 0 deaths, 5 deaths attributed to ketoacidosis, and 2 sudden deaths), and non-significantly (p = 0.16) decreased for macrovascular causes (3 vs 8 deaths). We conclude that there is a substantial risk of severe adverse effects associated with intensified insulin treatment. Mortality from acute metabolic causes is increased; however, this is largely counterbalanced by a reduction in cardiovascular mortality. The excess of severe hypoglycemia in the DCCT is not exceptional. Multiple daily injection schemes may be safer than treatment with insulin pumps. PMID- 9400916 TI - Symptomatic and physiological responses to hypoglycaemia induced by human soluble insulin and the analogue Lispro human insulin. AB - The aim of this study was to compare the glycaemic threshold for onset of the clinically detectable sympatho-adrenal (autonomic) reaction (defined as 'R') to hypoglycaemia induced by Lispro human insulin, with that induced by human soluble insulin (HS). The hypoglycaemia symptom profile, counterregulatory hormonal responses, and cognitive performance at R were also compared. Sixteen patients with IDDM, aged 32.5 (20-45) (median (range)) years and duration of IDDM 3.0 (0.5 4.5) years participated in a randomized, double-blind study during which intravenous infusions of either Lispro or HS insulin (2.0 mU kg(-1) min(-1)) were used on separate occasions to lower the blood glucose to the level at which R was induced. For both HS and Lispro, significant increments in systolic blood pressure (p<0.05), heart rate (p<0.05), and in autonomic (p<0.05) and neuroglycopenic symptom scores (p<0.05) occurred at R, and a significant deterioration was observed in cognitive performance (p<0.05). In response to hypoglycaemia, a significant increase from baseline occurred in plasma concentrations of all of the counterregulatory hormones (p<0.01) and the magnitude of response and temporal pattern did not differ, nor were any significant differences apparent between HS and Lispro insulins for any of the variables studied. The autonomic reaction occurred at a blood glucose (mean +/- SD) of 2.0 (+/- 0.6) mmol(-1) for Lispro and 1.9 (+/- 0.6) mmol(-1) for HS, which did not differ significantly. Thus, at the autonomic reaction to hypoglycaemia no significant differences were evident in the glycaemic threshold, symptom profile, physiological responses, and counterregulatory hormonal responses between Lispro and human soluble insulin in IDDM patients. PMID- 9400917 TI - Plasma leptin in non-diabetic Asian Indians: association with abdominal adiposity. AB - Plasma leptin concentrations were measured in 144 non-diabetic men and women (age 21-73 years, BMI 14.8-37.7 kg m(-2)), in fasting samples collected during a population survey for diabetes mellitus. Leptin, fasting and 2-h post-glucose load plasma concentrations of glucose and immunoreactive insulin were measured. In a subset of 50 normoglycaemic individuals, subcutaneous fat (SF) and visceral fat (VF) areas at L4-L5 level were also measured by CT. As in other populations, women had significantly higher plasma leptin concentrations than men (p < 0.001) but the values were similar in normal (NGT) and impaired glucose tolerance (IGT). Geometric mean concentrations of leptin in men and women with NGT were 4.8 and 17.7 ng ml(-1), respectively, and the corresponding values in IGT were 6.2 and 19.0 ng ml(-1). Multiple regression analysis in the total group showed that the leptin concentration (log-transformed) was strongly dependent on sex (R2 = 53.4%), BMI (R2 = 17.4%), and to a lesser degree on the 2-h plasma insulin (R2 = 2.4%) and the WHR (R2 = 0.8%). In men, the total abdominal fat showed a strong association with leptin (R2 = 49.3%) and in women the subcutaneous fat area showed a similar effect (R2 = 39.5%). It is likely that subcutaneous and not visceral fat may be a determinant of plasma leptin in Asian Indians, and the correlation between leptin and insulin resistance may be less strong than in other ethnic groups. PMID- 9400918 TI - Is an exaggerated postprandial triglyceride response associated with the component features of the insulin resistance syndrome? AB - To investigate whether individual component features of the insulin resistance syndrome were associated with the postprandial triglyceride response, 57 healthy Caucasian men between 57 and 70 years of age underwent a fat tolerance test lasting 8 h. Fasting triglyceride concentrations were associated with the total unfractionated postprandial triglyceride response (r(s) = 0.54, p < 0.001) and the triglyceride-rich lipoprotein (TGRLP) fraction (d < 1.006) at 8 h was associated with the maximum non-esterified fatty acid concentration (NEFA) (r(s) = 0.33, p = 0.01). Measures of obesity (BMI and WHR) were not associated with the postprandial triglyceride response but were inversely related to NEFA suppression (NEFA nadir and BMI, r(s) = 0.31, p = 0.02; and NEFA nadir and WHR, r(s) = 0.36, p = 0.006). Other component features of the IRS, including glucose tolerance and two proxy measures of insulin resistance (fasting insulin concentration and HOMA measurement) were not associated with the postprandial triglyceride response despite being strongly associated with fasting triglyceride concentration. Current smoking habit, chronic alcohol consumption and birth weight were also not associated with an altered postprandial triglyceride response. In conclusion these results show that although component features of the IRS were associated with increased fasting triglyceride concentrations many of these features, including two proxy measures of insulin sensitivity were not associated with an exaggerated postprandial triglyceride response. PMID- 9400919 TI - A 3-19-year follow-up study on diabetic retinopathy in patients diagnosed in childhood and treated with conventional therapy. AB - Few data are available from follow-up studies on diabetic retinopathy in patients diagnosed with insulin-dependent (Type 1) diabetes mellitus in childhood and treated with conventional therapy. We report the results of conventional insulin therapy on development of diabetic retinopathy in 100 children and adolescents (47 females and 53 males), aged 8.3 +/- 3.5 (1.2-16.4) years at diagnosis of disease. Oral or intravenous fluorescein angiography was performed during a 3-19 year follow-up in all patients. Retinopathy was staged according to the criteria of the Italian Society of Diabetology (SID). During follow-up, retinopathy was observed in 28 patients (28%). At the end of follow-up, retinopathy was present in 23 patients and had disappeared in 5. Life-table analysis showed a median disease-free interval of 10.8 years. At 10 years from diagnosis the percentage of patients free of retinopathy was 66%. Poor metabolic control, age, and degree of pubertal development at diagnosis were the most important risk factors. PMID- 9400920 TI - Effect of insulin on blood rheology in non-diabetic subjects and in patients with Type 2 diabetes mellitus. AB - We evaluated the effect of insulin on platelet function, blood viscosity, and filterability in healthy subjects and in patients with Type 2 (non-insulin dependent) diabetes mellitus. Fifteen diabetic patients were free from cardiovascular complications (group A), while the other 15 patients had both clinical and measured evidence of coronary or peripheral vascular disease (group B); 15 non-diabetic subjects served as controls. On blood samples taken without stasis, maximal platelet aggregation to 1.25 micromol l(-1) ADP, blood and plasma viscosity, and blood filterability were measured in basal conditions, and after incubation of blood, plasma or platelet-rich plasma with insulin at two physiological concentrations (120 and 480 pmol l(-1)). Compared with healthy subjects, the diabetic patients of group B had higher values of blood (p < 0.01) and plasma (p < 0.05) viscosity, and platelet aggregation response to ADP (p < 0.01), as well as lower values of blood filterability (p < 0.01). The diabetic patients of group A had values intermediate between normal subjects and the patients of group B. In non-diabetic subjects, insulin significantly decreased platelet aggregation and blood viscosity at low shear rates (22.5 s(-1)) (p < 0.01 for both), and had no significant effects on other parameters. In the diabetic patients of group A, insulin decreased blood viscosity at high (225 s( 1)) rates of shear (p < 0.01) and increased blood filterability (p < 0.01). The effects of insulin were not dose-related. In the diabetic patients of group B, none of the parameters evaluated was significantly influenced by insulin. Type 2 diabetic patients present many abnormalities of the rheologic properties of blood. The beneficial effects of insulin on platelet aggregation and blood viscosity are not evident in Type 2 diabetic patients, especially those with vascular complications and this may be relevant to the development of those complications. PMID- 9400921 TI - Comparison of incidence of insulin-dependent diabetes mellitus in children and young adults in the Province of Turin, Italy, 1984-91. Piedmont Study Group for Diabetes Epidemiology. AB - To document the incidence of IDDM in the Province of Turin (Italy) in the 8-year period 1984-91 in children (0-14 years) and young adults (15-29 years), in relation to age, sex, monthly-seasonal variability, calendar year and urban/rural area, (all newly diagnosed cases (502) were ascertained through primary and secondary data sources and completeness of ascertainment estimated with the two sample capture-recapture method (99% in childhood and 95% in young adults). The independent effect of age, sex, calendar year, and urban/rural area was estimated with a Poisson regression model. Age-specific incidence rates were 8.42/100,000 (95% CI 7.37-9.62) and 6.72/100,000 (95% CI 5.96-7.58), respectively, in the age groups 0-14 and 15-29 years. Sex differences were evident in young adults, with an almost 1.5-fold increased risk in men (8.37/100,000, 95% CI 7.21-9.71 vs 5.00/100,000, CI 4.09-6.10). Seasonal trend was evident in childhood. Predictors of incidence rates were age, place of residence and interaction between sex and age; no temporal trend was detected. No significant differences were found in the two age-groups with respect to glycaemia, glycosuria, ketonuria, and fasting C peptide levels. In conclusion, this study shows sex differences in IDDM risk in young adults; 55% of incident cases occurring in young adults; an independent contribution of urban/rural differences to IDDM risk; no temporal trend in 1984 91; a seasonal pattern of incidence in children; no significant differences in clinical presentation between age groups. PMID- 9400922 TI - Can digitised colour 35 mm transparencies be used to diagnose diabetic retinopathy? AB - Retinal photography is an adjunct to ophthalmoscopy in screening for diabetic retinopathy (DR). Digital retinal cameras allow a retinal image to be displayed immediately on a high resolution video display monitor. We conducted a pilot study to investigate the agreement in retinopathy grading from digitized images in comparison to original colour transparencies as 35 mm slides. One hundred and fifty macula-centred, 45 degree, non-stereoscopic retinal images were digitized onto CD ROM by Kodak at base resolution of 768 x 512 pixels. The anonymized images were displayed on a 17" monitor running Windows at 800 x 600 resolution in 64,000 colours (PC images) and graded in random order. Alternatively the transparencies were graded on a Slidex viewer. A quality control set were also graded with exact agreement in 93% of cases (91% (73/80) of PC images and 94% (75/80) of slide images). Compared to colour transparencies, 95% (84/88) of sight threatening diabetic retinopathy (STDR) and 100% (62/62) of non-STDR cases were diagnosed using the PC. One case of pre-proliferative DR and three cases of non proliferative DR were graded as non-STDR from the PC. There was good agreement between PC displayed digitized retinal images and 35 mm colour transparencies. PMID- 9400923 TI - Low-density lipoprotein size, high-density lipoprotein concentration, and endothelial dysfunction in non-insulin-dependent diabetes. AB - We examined endothelial function (nitric-oxide mediated) in 29 men with diet treated non-insulin-dependent (Type 2) diabetes mellitus (NIDDM) and 18 male age matched controls. Forearm blood flow was measured by venous occlusive plethysmography during intra-arterial administration of acetylcholine (ACh, 7.5 and 15 microg min(-1)) and sodium nitroprusside (SNP, 3 and 10 microg min(-1)). LDL particle size was estimated by non-denaturing gel electrophoresis. Serum lipids, blood pressure, and glycated haemoglobin were also measured. LDL particle size was smaller (p = 0.048) in the diabetic patients than controls. In the diabetic patients, LDL particle size was a significant positive predictor (p = 0.01) of the area under the dose-response curve for ACh, after adjusting for age, HbA1c, systolic BP, and cholesterol (R2 0.20). In stepwise regression including serum lipid and lipoprotein concentrations and LDL particle size, decreased HDL cholesterol was the best predictor of an impaired vasodilatory response to ACh. Vasodilatory responses to sodium nitroprusside were not significantly correlated with LDL particle size or serum lipid and lipoprotein concentrations. We conclude that in men with NIDDM, small, dense LDL particle size is associated with abnormal endogenous release of nitric oxide. The contribution of small, dense LDL particles to the development of endothelial dysfunction and early diabetic vasculopathy may not, however, be as great as decreased HDL cholesterol. PMID- 9400924 TI - Return of beta-adrenergic sensitivity in a patient with insulinoma after removal of the tumour. AB - Beta-adrenergic sensitivity and counterregulatory hormone and symptomatic responses to hypoglycaemia were studied in a 22-year-old man before and 3 and 34 weeks after removal of an insulinoma. The beta-adrenergic sensitivity was measured by the effect of an isoprenaline infusion on the heart rate, and the dose needed to increase the heart rate by 25 beats min(-1) (I25) calculated from regression lines. The glucose thresholds for the hormonal responses and symptoms were studied during a gradual fall in plasma glucose using a hypoglycaemic clamp technique. As compared with preoperative values, beta-adrenergic sensitivity was unchanged 3 weeks after surgery, but showed a marked improvement after 34 weeks, the I25 (in microg isoprenaline) being 0.96, 0.86, and 0.56, respectively. The hormone responses to hypoglycaemia were earlier, but with no improvement in symptom generation at 3 weeks. After 34 weeks, the thresholds for both hormone release and symptom generation occurred at a plasma glucose approximately 1 mmol l(-1) higher than before surgery. Thus, in our patient, there was a marked improvement in beta-adrenergic sensitivity, an earlier release of counterregulatory hormones, and an earlier recognition of hypoglycaemic symptoms after surgery. However, the restoration of these responses took more than 3 weeks. PMID- 9400926 TI - Diabetes information in the Cochrane Library. PMID- 9400925 TI - Selective intra-arterial calcium injection in the investigation of adult nesidioblastosis: a case report. AB - A 69-year-old man with recurrent hypoglycaemia had inappropriately elevated plasma insulin level during a symptomatic hypoglycaemia, but had a negative prolonged fast. Computerized tomography (CT) of the abdomen revealed a nodular lesion over the body of pancreas, whereas pancreatic arteriography failed to show tumour blush. Hence, arterial stimulation (with calcium) and venous sampling (ASVS) was performed and a brisk response of plasma insulin level was found when calcium was injected both into the splenic and the superior mesenteric arteries. Since no tumour was found during the operation, the patient received subtotal distal pancreatectomy. Pathological examination of the resected tissue disclosed a typical finding of nesidioblastosis. We suggest that selective intra-arterial calcium injection with hepatic venous sampling for insulin gradients is useful for the diagnosis of adult nesidioblastosis. PMID- 9400927 TI - Ketosis-onset diabetes in young adults with subsequent non-insulin-dependency, a link between IDDM and NIDDM? PMID- 9400929 TI - In defence of twin studies: look to the phenotype. PMID- 9400928 TI - Soluble selectins in IDDM: possible association with lipids? PMID- 9400930 TI - Poor glycaemic control and retinopathy in non-insulin-dependent diabetes mellitus. PMID- 9400931 TI - A miracle of salamization: positive divided by two equals negative. PMID- 9400932 TI - Combined inhibition of topoisomerases I and II--is this a worthwhile/feasible strategy? PMID- 9400933 TI - Circulating soluble adhesion molecules E-cadherin, E-selectin, intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in patients with gastric cancer. AB - The concentrations of the soluble adhesion molecules E-cadherin, E-selectin, intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) were investigated in 45 patients with gastric cancer before treatment and their correlation with clinical, histological and routine laboratory parameters was examined. Data were collected on tumour stage at presentation, presence and sites of metastatic disease, tumour pathology, survival and results of routine laboratory tests. Serum concentrations of ICAM-1 and VCAM-1 were significantly elevated in the patients with gastric cancer in comparison with the group of healthy subjects (P < 0.00001 and P < 0.0001 respectively). Increased serum concentrations of VCAM-1 were associated with locally advanced and metastatic disease whereas ICAM-1 was significantly elevated both in local and in advanced/metastatic disease. Soluble E-cadherin and E-selectin concentrations did not show any significant elevation in gastric cancer patients. Concentrations of soluble adhesion molecules showed significant correlation with each other (except E-selectin and VCAM-1) and with alkaline phosphatase. Soluble ICAM-1 and VCAM-1 were significantly associated with an elevated total white cell count. Patients with elevated VCAM-1 had significantly poorer survival in comparison with patients with normal serum levels (P = 0.0361). PMID- 9400934 TI - The RB1 gene mutation in a child with ectopic intracranial retinoblastoma. AB - The RB1 gene mutation was investigated in a child with ectopic intracranial retinoblastoma using DNA obtained from both the pineal and retinal tumours of the patient. A nonsense mutation in exon 17 (codon 556) of the RB1 gene was found to be present homozygously in both the retinal and the pineal tumours. The same mutation was present heterozygously in the DNA from the constitutional cells of the patient, proving it to be of germline origin. The initial mutation was shown to have occurred in the paternally derived RB1 allele. The mutation is in an area of the gene that encodes the protein-binding region known as the 'pocket' region and has been detected in other cases of retinoblastoma. PMID- 9400935 TI - Angiogenesis is associated with vascular endothelial growth factor expression in cervical intraepithelial neoplasia. AB - Squamous cell carcinoma of the cervix (SCC) is preceded by a premalignant condition known as cervical intraepithelial neoplasia (CIN). The majority of cases of CIN regress spontaneously; however, methods are needed to identify those lesions likely to progress. Increased blood vessel density, signifying angiogenesis, is an independent prognostic indicator in a number of cancers, although little is known about its significance in premalignant lesions. The aim of the present study was to determine the relationship between vessel density, expression of the potent angiogenic factor vascular endothelial growth factor (VEGF) and CIN grade. Using immunohistochemistry, mean vessel density (MVD) and VEGF expression were assessed in samples from 54 patients who had undergone cone biopsy for CIN or hysterectomy for SCC and from 16 patients with no cervical pathology. There were significant increases in MVD and VEGF expression from normal cervix through CIN I to CIN III to invasive SCC, but no difference in mean vessel diameter between groups. There was a strong correlation between mean vessel density and VEGF expression, and both were associated with histological grade of CIN. The original MVDs for a small group of patients later presenting with recurrent disease were found to be equal to or greater than the mean for their histological grade. We conclude that the onset of angiogenesis is an early event in premalignant changes of the cervix due, in part, to enhanced expression of VEGF by the abnormal epithelium. PMID- 9400936 TI - Prognostic value of loss of heterozygosity at BRCA2 in human breast carcinoma. AB - To confirm several recent studies pointing to loss of heterozygosity (LOH) at BRCA2 as a prognostic factor in sporadic breast cancer, we examined this genetic alteration in a large series of human primary breast tumours for which long-term patient outcomes were known. LOH at BRCA2 correlated only with low oestrogen and progesterone receptor content. Univariate analysis of metastasis-free survival and overall survival (log-rank test) showed no link with BRCA2 status (P = 0.34, P = 0.29 respectively). LOH at BRCA2 does not therefore appear to be a major prognostic marker in sporadic breast cancer. PMID- 9400937 TI - Down-regulation of a novel form of fibroblast growth factor receptor 1 in human breast cancer. AB - Monoclonal antibodies against two epitopes of FGFR-1 have been used to investigate FGFR-1 expression in the normal and neoplastic human breast. Different forms are detected in the different cell types constituting the normal breast. Moreover, breast cancer cells lack one form of FGFR-1. Western blot analysis showed 115-kDa and 106-kDa forms of FGFR-1 within the human breast. The 115-kDa band corresponds to the beta form of FGFR-1, whereas the 106-kDa band is truncated at the carboxyl terminus. The 106-kDa form of FGFR-1 is the major form present in breast fibroblasts and myoepithelial cells, whereas epithelial cells contain equal amounts of the 115-kDa and 106-kDa forms. Breast cancer cells, however, appear to contain only the 115-kDa form of FGFR-1. This expression pattern is reflected in malignant and non-malignant tissue samples. Using reverse transcription polymerase chain reaction (RT-PCR) analysis, we have shown that the 106-kDa FGFR-1 isoform is not the previously described alpha 2 receptor that arises from a 25-base pair insertion in the second kinase domain. It is probable that the 106-kDa FGFR-1 has different signalling properties to the full-length receptor, having lost at least one tyrosine at amino acid 766, which is required for phospholipase C activation. This form of FGFR-1 appears to be lost in all breast cancer cells analysed and its absence may have a bearing on malignancy. PMID- 9400938 TI - Germline BRCA2 mutations in men with breast cancer. AB - Breast cancer in men is rare and is clearly due in some cases to an inherited predisposition. A total of 28 male breast cancer patients were tested for BRCA2 mutations; two frameshifts and one putative missense mutation were identified. One of the frameshifts was detected in the same position as a mutation estimated to be responsible for 40% of all male breast cancer cases in Iceland. PMID- 9400939 TI - The effect of hypoxia and hyperoxia on nucleoside triphosphate/inorganic phosphate, pO2 and radiation response in an experimental tumour model. AB - This study has evaluated the effect of breathing 100% oxygen, carbogen and carbon monoxide (at 660 p.p.m.) on the bioenergetic and oxygenation status and the radiation response of 200-mm3 C3H mammary carcinomas grown in the feet of CDF mice. Bioenergetic status was assessed by 31P magnetic resonance spectroscopy (MRS) using a 7-tesla spectrometer with both short (2 s) and long (6 s) pulse repetition times. Tumour partial pressure of oxygen (PO2) was measured with an Eppendorf polarographic electrode; the oxygenation parameters were the median pO2 and fraction of pO2 values < or = 2.5 mmHg. The radiation response was estimated using a tumour growth delay assay (time to grow three times treatment volume). Carbon monoxide breathing decreased tumour pO2 and compromised the radiation response, but the beta-nucleoside triphosphate (NTP)/Pi ratio was unchanged. Both carbogen and oxygen (100%) increased tumour pO2 and beta-NTP/Pi and enhanced the radiation response, the effects being similar under the two gassing conditions and dependent on the gas breathing time. Thus, in this tumour model, 31P-MRS can detect hyperoxic changes, but because cells can remain metabolically active even at low oxygen tensions the beta-NTP/Pi did not correlate with low tissue oxygenation. An analysis of variance showed that gas breathing time induced a significant systematic effect on beta-NTP/Pi, the MRS pulse repetition time had a significant effect on beta-NTP/Pi change under hypoxic but not under hyperoxic conditions and the type of gas that was inhaled had a significant effect on beta NTP/Pi. PMID- 9400940 TI - Differential level of DSB repair fidelity effected by nuclear protein extracts derived from radiosensitive and radioresistant human tumour cells. AB - A cell-free plasmid reactivation assay was used to determine the fidelity of DNA double-strand break (DSB) repair in a panel of eight DSB repair-proficient human tumour cell lines. Nuclear protein extracts derived from radiosensitive tumour cells were less capable of correctly rejoining EcoRI-induced DSBs than were similar extracts from radioresistant tumour cells. Linear regression analysis suggests that there was a significant (r2 = 0.84, P = 0.001, d.f. = 6) correlation between the fidelity of DSB rejoining and the SF2 values of the cell lines studied. This cell-free assay is clearly sensitive to differences in the nuclear protein composition that reflect the clinically relevant radiosensitivity of these cell lines. The fact that our cell-free assay yielded similar results to previous studies that used intracellular plasmid reactivation assays suggests that those differences in DSB mis-rejoining frequencies in radiosensitive and radioresistant cell lines may be due to inherent differences in nuclear protein composition and are not directly attributable to differences in proliferation rates between cell lines. The underlying cause for this association between DSB mis-rejoining frequencies and radiosensitivity is presently unknown, however restriction endonuclease mapping and polymerase chain reaction (PCR) amplification analysis revealed that approximately 40% of the mis-rejoined DSBs arose as a result of the deletion of between 40 and 440 base pairs. These data raise the possibility that the radiosensitivity of DSB repair-proficient human tumour cell lines may be partly determined by the predisposition of these cell lines to activate non-conservative DSB rejoining pathways. PMID- 9400941 TI - Similarity of apoptosis induction by 2-chlorodeoxyadenosine and cisplatin in human mononuclear blood cells. AB - The purine analogue 2-chlorodeoxyadenosine (CdA) is unique compared with traditional antimetabolite drugs, as it has shown equal activity in dividing and resting lymphocytes. Poly(ADP-ribose)polymerase (PARP) activation and consecutive NAD+ consumption have been associated with the induction of apoptosis in resting cells. The potential of CdA to induce the p53-dependent DNA damage response was assessed in resting and phytohaemagglutinine (PHA)-activated peripheral blood mononuclear cells (PBMCs) and compared with cisplatin (DDP), a cell cycle dependent and DNA-damaging agent that is mainly used in the treatment of solid tumours. Both drugs induced transactivation of the p53 target genes waf1 and mdm2, NAD+ consumption and apoptotic death. The expression pattern of p53 and waf1 suggests a partly p53-independent induction of waf1. The expression of c-myc and PARP, which both have a dual role in proliferation and apoptosis, was selectively induced by CdA. Cell cycle stimulation increased the cytotoxic activity of both drugs. These data show that DDP is also a potent inducer of apoptosis in resting and proliferating peripheral blood mononuclear cells. Activation of the p53-dependent DNA damage response seems to be an important component of the toxic effect of CdA. PMID- 9400942 TI - Allelotype analysis of adenocarcinoma of the gastric cardia. AB - To identify chromosomal loci involved in the development of proximal gastric adenocarcinoma, this study delineated the pattern of allelic imbalance in a series of 38 adenocarcinomas arising in the gastric cardia. A total of 137 microsatellite markers covering all autosomal arms, excluding acrocentric arms, were analysed. A mean of 35 out of a total of 39 chromosomal arms studied were informative for each patient. The tumour group demonstrated a high level of allelic imbalance, with an observed median fractional allelic imbalance of 0.47 for the 29 intestinal-type adenocarcinomas and 0.54 for the nine diffuse-type adenocarcinomas. Allelic imbalance was detected in >50% of informative cases in both histological subtypes on a number of chromosomal arms. In the intestinal subtype, these included, 3p (61%), 4q (71%), 5q (59%), 8p (60%), 9p (65%), 9q (83%), 12q (52%), 13q (52%), 17p (78%) and 18q (70%). A higher incidence of allelic imbalance was detected on chromosome 16q in tumours of the diffuse type relative to those of the intestinal type. A more detailed mapping on chromosomes 4q and 6q identified a number of cases with subchromosomal breakpoints. In conclusion, this analysis has indicated regions of the genome potentially involved in the development of proximal gastric carcinomas. PMID- 9400943 TI - Platinum-DNA adduct formation in leucocytes of children in relation to pharmacokinetics after cisplatin and carboplatin therapy. AB - Platinum (Pt)-DNA adducts were measured in peripheral blood leucocytes (PBLs) from 24 children with solid tumours after standard cisplatin and/or carboplatin treatment. The relationship between Pt-DNA adduct levels and pharmacokinetics of cisplatin and carboplatin was investigated. Adduct measurements were performed by competitive enzyme-linked immunosorbent assay (ELISA) and plasma unbound Pt concentrations were measured by atomic absorption spectrophotometry (AAS). There was considerable interindividual variation in Pt-DNA adduct level that was weakly correlated (r2 = 0.32) with the area under the unbound drug concentration vs time curve (AUC) at 6 h after the start of cisplatin infusion, indicating that the variation in Pt-DNA adduct levels was primarily determined by factors other than AUC. No clear relationship between AUC and adduct levels was seen at 24 and 48 h after cisplatin or at 6, 24 or 48 h after carboplatin. Carboplatin produced lower levels of immunoreactive adducts than did cisplatin (1.3 +/- 0.6 nmol Pt g-1 DNA vs 3.2 +/- 1.7 nmol Pt g-1 DNA), despite a 20-fold higher unbound drug AUC for carboplatin (8.0 +/- 3.5 mg ml-1 min vs 0.4 +/- 0.2 mg ml-1 min). This study demonstrates that, after cisplatin and carboplatin treatment the drug-target interaction is determined by both pharmacokinetic and, predominantly, cellular factors. Intrinsic differences between the two complexes, primarily reactivity, probably explain the lower adduct levels observed after carboplatin treatment. PMID- 9400944 TI - A phase I/II study of a hypoxic cell radiosensitizer KU-2285 in combination with intraoperative radiotherapy. AB - A fluorinated 2-nitroimidazole radiosensitizer KU-2285 was given before intraoperative radiotherapy (IORT) to 30 patients with unresectable, unresected or macroscopic residual tumours. Twenty-three patients had pancreatic cancer and five had osteosarcoma. The IORT dose was 30 Gy for unresectable pancreatic cancer and 60 Gy for osteosarcoma. The dose of KU-2285 administered ranged from 1 to 9 g m-2. Four patients received a dose of 9 g m-2, and ten received 6.8-7 g m-2. All patients tolerated KU-2285 well, and no drug-related toxicity was observed. The average tumour concentration of KU-2285 immediately after IORT was 166 microg g-1 at dose of 6.8-7 g m-2 and 333 microg g-1 at 9 g m-2. The average tumour-plasma ratio was > or = 0.82. Eleven patients with unresectable but localized pancreatic cancer treated with KU-2285 plus IORT and external beam radiotherapy had a median survival time of 11 months and 1-year local control rate of 50%, which compares favourably with those of 8 months (P = 0.26) and 28% (P = 0.10) for 22 matched historical control patients. The five patients with osteosarcoma attained local control. The results of this first study on KU-2285 and IORT appear encouraging, and further studies of this compound seem to be warranted. PMID- 9400945 TI - Conversion of the prodrug etoposide phosphate to etoposide in gastric juice and bile. AB - Etoposide phosphate is a water-soluble prodrug of etoposide. It was expected that this prodrug could be used to overcome the solubility limitations and erratic bioavailability of oral etoposide. To investigate the possibility of prodrug conversion to etoposide within the gastrointestinal lumen, etoposide phosphate was dissolved in water and incubated with human gastric juice or human bile in vitro. Samples were collected during 150 min and analysed for etoposide concentration with high-performance liquid chromatography. Conversion of prodrug to etoposide during incubation with gastric juice was negligible. There was significant conversion during incubation with bile at pH 7-8. The percentage of prodrug converted to etoposide at pH 8 after 60 min was 78 +/- 18% (mean +/- S.D.) for a 0.1 mg ml-1 prodrug solution and 36 +/- 26% for 0.5 mg ml-1. At pH 7, after 60 min 22% of prodrug was converted to etoposide when incubated at 0.1 mg ml-1 and 10% at 0.5 mg ml-1. No conversion was found after inactivation of alkaline phosphate (AP) by overnight heating of bile at 65 degrees C or by the addition of disodium edetate to the bile. In conclusion, because of AP in bile, variable conversion of etoposide phosphate to etoposide can be expected within the intestinal lumen after oral administration. This could have important pharmacokinetic consequences. PMID- 9400946 TI - A prospective study of surgery and adjuvant chemotherapy for primary gastric lymphoma stage II. AB - The standard management of primary gastric lymphoma (PGL) (stage II) has not been established despite the use of various treatment modalities. The present prospective trial of combined surgery and chemotherapy for the treatment of PGL (stage II) included 25 consecutive patients treated between July 1978 and December 1993. Twenty-one patients were treated with total gastrectomy and four with partial gastrectomy; this was followed by post-operative chemotherapy with m VEPA (vincristine, cyclophosphamide, prednisolone and doxorubicin), followed by consolidation chemotherapy with VEMP (vindesine, cyclophosphamide, methotrexate and prednisolone) or VQEP (vindesine, carbazilquinone, cyclophosphamide and prednisolone). Twenty-one of the 25 patients who completed post-operative chemotherapy were free of relapse 26-203 (median 94) months after the gastrectomy. Of the four patients who did not complete the projected chemotherapy, two relapsed and died of lymphoma. Another patient with recurrent lymphoma died in an accident, and the fourth patient was in remission at 54 months after surgery. The post-operative overall and disease-free survival rates at 10 years for the 25 evaluable patients were 81.6% and 92.0% respectively. Major surgical complications and treatment-related death after chemotherapy were not observed. PGL (stage II) appears to be curable when treated with gastrectomy and adjuvant chemotherapy. PMID- 9400947 TI - Phase I study of gemcitabine using a once every 2 weeks schedule. AB - Gemcitabine (2',2'-difluorodeoxycytidine) is a novel nucleoside analogue. As part of a series of studies to determine the maximum tolerated dose (MTD) of gemcitabine and the most appropriate schedule, a two-centre phase I study of gemcitabine was undertaken in patients with advanced refractory solid tumours using a once every 2 weeks schedule. Fifty-two patients were entered into the study at 14 different dose levels (40-5700 mg m-2). Weekly evaluations for toxicity were performed and the MTD for this once every 2 weeks schedule was 5700 mg m-2. The dose-limiting toxicity was myelosuppression, with neutropenia being most significant. Other toxicities were nausea, vomiting, fever and asthenia. One minor response was seen in a heavily pretreated breast cancer patient treated at 1200 mg m-2. Preclinical studies suggest that the efficacy of gemcitabine is more schedule than dose related, and it is concluded that this is not the most appropriate dosing schedule for gemcitabine. However, this study demonstrates the safety profile of gemcitabine, as doses over fourfold greater than that recommended for the weekly schedule of 1000 mg m-2 could be tolerated. PMID- 9400948 TI - Phase I study of sequentially administered topoisomerase I inhibitor (irinotecan) and topoisomerase II inhibitor (etoposide) for metastatic non-small-cell lung cancer. AB - We conducted a phase I study of irinotecan (CPT-11) and etoposide (VP-16) given sequentially to untreated patients with metastatic non-small-cell lung cancer. Arm A: CPT-11 was given over 90 min on days 1-3 and VP-16 was given over 60 min on days 4-6. Arm B: VP-16 was given on days 1-3 and CPT-11 on days 4-6. G-CSF was given to all patients daily on days 7-17. Twenty-seven patients were entered randomly at the two arms. The major dose-limiting toxicities in arms A and B were granulocytopenia and diarrhoea. Transient elevations of transaminases and bilirubin were observed in both arms. The degree of the toxicities did not differ between the two arms. The maximum tolerated doses (MTDs) were 60 mg m-2 CPT-11 and 60 mg m-2 VP-16 in both arms. Of the 13 patients who received more than two cycles, two out of five achieved partial response (PR) at the first level of arm A and one out of four achieved PR at the second level of arm B. We conclude that these schedules of sequential CPT-11 and VP-16 administration were inappropriate because of severe toxicities. PMID- 9400949 TI - Phase I and pharmacological study of sequential intravenous topotecan and oral etoposide. AB - We performed a phase I and pharmacological study to determine the maximum tolerated dose (MTD) and dose-limiting toxicities (DLT) of a cytotoxic regimen of the novel topoisomerase I inhibitor topotecan in combination with the topoisomerase II inhibitor etoposide, and to investigate the clinical pharmacology of both compounds. Patients with advanced solid tumours were treated at 4-week intervals, receiving topotecan intravenously over 30 min on days 1-5 followed by etoposide given orally twice daily on days 6-12. Topotecan-etoposide dose levels were escalated from 0.5/20 to 1.0/20, 1.0/40, and 1.25/40 (mg m-2 day 1)/(mg bid). After encountering DLT, additional patients were treated at 3-week intervals with the topotecan dose decreased by one level to 1.0 mg m-2 and etoposide administration prolonged from 7 to 10 days to allow further dose intensification. Of 30 patients entered, 29 were assessable for toxicity in the first course and 24 for response. The DLT was neutropenia. At doses of topotecan etoposide 1.25/40 (mg m-2)/(mg bid) two out of six patients developed neutropenia grade IV that lasted more than 7 days. Reduction of the treatment interval to 3 weeks and prolonging etoposide dosing to 10 days did not permit further dose intensification, as a time delay to retreatment owing to unrecovered bone marrow rapidly emerged as the DLT. Post-infusion total plasma levels of topotecan declined in a biphasic manner with a terminal half-life of 2.1 +/- 0.3 h. Total body clearance was 13.8 +/- 2.7 l h-1 m-2 with a steady-state volume of distribution of 36.7 +/- 6.2 l m-2. N-desmethyltopotecan, a metabolite of topotecan, was detectable in plasma and urine. Mean maximal concentrations ranged from 0.23 to 0.53 nmol l-1, and were reached at 3.4 +/- 1.0 h after infusion. Maximal etoposide plasma concentrations of 0.75 +/- 0.54 and 1.23 +/- 0.57 micromol l-1 were reached at 2.4 +/- 1.2 and 2.3 +/- 1.0 h after ingestion of 20 and 40 mg respectively. The topotecan area under the plasma concentration vs time curve (AUC) correlated with the percentage decrease in white blood cells (WBC) (r2 = 0.70) and absolute neutrophil count (ANC) (r2 = 0.65). A partial response was observed in a patient with metastatic ovarian carcinoma. A total of 64% of the patients had stable disease for at least 4 months. The recommended dose for use in phase II clinical trials is topotecan 1.0 mg m-2 on days 1-5 and etoposide 40 mg bid on days 6-12 every 4 weeks. PMID- 9400951 TI - The role of occupation and a past history of malaria in the etiology of classic Kaposi's sarcoma: a case-control study in north-east Sardinia. AB - A case-control study was performed to determine the role of rural factors including occupation and previous malaria exposure in the development of classic Kaposi's sarcoma (CKS) in a high incidence area of Europe. The occurrence of CKS association with other malignancies was also examined. The results showed that the risk of having CKS was significantly increased in subjects farming cereals, while a previous history of malaria did not influence the risk of developing CKS. A near-significant increase in associated tumours was found. PMID- 9400950 TI - Cisplatin and vinorelbine followed by ifosfamide plus epirubicin vs the opposite sequence in advanced unresectable stage III and metastatic stage IV non-small cell lung cancer: a prospective randomized study of the Southern Italy Oncology Group (GOIM). AB - A multicentric, prospective phase III study was carried out with the aim of testing the so-called 'worst drug rule' hypothesis, which suggests the use of an effective but 'less active' regimen that first eradicates tumoral cells resistant to a second effective and 'more active' regimen. With respect to this hypothesis, we considered the cisplatin plus vinorelbine regimen (CCDP/VNR) as the more active regimen compared with the non-cisplatin-containing regimen of ifosfamide plus high-dose epirubicin (IFO/EPI). Thus, a randomized study was carried out to compare the sequencial strategy of three cycles of CDDP/VNR followed by three cycles of IFO/EPI with the opposite sequence in advanced non-small-cell lung cancer. A total of 100 consecutive previously untreated patients with stage III IV non-small-cell lung cancer were centrally randomized in two arms according to stage of disease and the performance status. Patients allocated to arm A received CDDP (100 mg m-2 on day 1) plus VNR (25 mg m-2 i.v. on days 1 and 8) every 21 days for three cycles (step 1) followed, after restaging, by three cycles of IFO (2.5 g m-2 with mesna on day 1) plus high-dose EPI (100 mg m-2 on day 1) every 21 days (step 2). Patients in arm B received the opposite sequence. Type and rates of objective response were evaluated after step 1 and step 2 in agreement with WHO criteria and an intent-to-treat analysis. Patients were also analysed for toxicity patterns, time to progression and survival. After the first three cycles (step 1), overall response rate (ORR), calculated according to an intent-to-treat analysis, was 47% and 21% for arm A and arm B respectively (P = 0.0112). ORR for stage III patients was 55% and 14% for arm A and B respectively (P = 0.0097). In stage IV patients ORR was higher in arm A than in arm B (42% vs 28%) but not statistically significant (P = 0.4). Clinical responses to the shift of chemotherapy (step 2) showed that no patient pretreated with CDDP/VNR and subsequently treated with IFO/EPI showed further response, whereas in the inverse sequence arm CDDP/VNR was able to induce 26% partial response (PR) rate in patients pretreated with IFO/EPI. This difference was statistically significant (P = 0.037). The overall median time to progression (TTP) of arm A and arm B did not significantly differ (6 vs 4 months; P = 0.665). However, median TTP of stage III patients was, respectively, 7 months for arm A and only 3 months for arm B. This difference was statistically significant (P = 0.049). Median overall survival (OS) was 9 and 7 months respectively for arm A and arm B. Despite this trend the difference was not significant (P = 0.328). Median OS of stage III patients showed a statistically significant advantage for arm A over arm B (13 vs 7 months, P = 0.03). In addition, no statistically significant difference in OS was recorded for stage IV patients (both arms 7 months, P = 0.526). Our data do not confirm Day's 'worst drug rule' hypothesis, at least in patients with advanced non-small-cell lung cancer treated with the above-mentioned regimens. The combination of CDDP and VNR seems more active, at least in terms of response rate, than the IFO/EPI, which performed poorly. PMID- 9400952 TI - Melanoma risk and residence in sunny areas. EORTC Melanoma Co-operative Group. European Organization for Research and Treatment of Cancer. AB - Melanoma risk among subjects from Germany, France and Belgium who had lived for 1 year or more in sunny climates was examined in a one-to-one unmatched case control study conducted among white subjects 20 years old or more. A total of 412 consecutive patients with melanoma diagnosed from 1 January 1991 onwards, were derived from hospital registers; 445 controls were randomly chosen in the same municipality as the cases. After adjustment for host characteristics, melanoma risk associated with residence in a sunny area was 2.7 (95% CI: 1.4-5.2), increasing to 4.7 (95% CI: 1.4-13.5) if subjects sought a suntan when residing in sunny climates, and to 4.3 (95% CI: 1.7-11.1) if subjects arrived before the age of 10 years in the sunny area. Residence in sunny areas and recreational sun exposure seemed to combine their effects on melanoma risk. Increase in melanoma risk conveyed by deliberate sun exposure during adulthood was highest among subjects who had lived in sunny areas as a child or adolescent and lowest among subjects who had never resided in sunny areas. Our results support conclusions from migrant studies that indicated that childhood is a critical period of either vulnerability to solar radiation or more frequent exposures to melanoma risk factors. They also suggest that moderate sun exposure of an adult who was heavily sun exposed in childhood is associated with a higher melanoma risk than that of high sun exposure of an adult who was sun protected in childhood. PMID- 9400953 TI - Childhood cancer and parental use of tobacco: deaths from 1971 to 1976. AB - Parental smoking data have been reabstracted from the interview records of the Oxford Survey of Childhood Cancers (deaths from 1971 to 1976). Reported smoking habits for the parents of 2587 children who died with cancer were compared with similar information for the parents of 2587 healthy controls (matched pairs analysis). Maternal daily consumption of cigarettes and paternal use of pipes or cigars were unimportant, but there was a statistically significant positive trend between paternal daily consumption of cigarettes and the risk of childhood cancer (P < 0.001). This association could not be explained by maternal smoking, social class, parental ages at the birth of the survey child, sibship position or obstetric radiography. Relations between maternal consumption of cigarettes and birth weights suggested that (maternal) smoking data were equally reliable for case and control subjects. About 14% of all childhood cancers in this series could be attributable to paternal smoking. These data were combined with smoking data from two previously published reports from the Oxford Survey (deaths from 1953 to 1955, deaths from 1977 to 1981) to obtain further information on risks for different types of cancer and different ages at onset of disease. Paternal cigarette smoking emerged as a potential risk factor both for the generality of childhood cancer and for all ages at onset. PMID- 9400955 TI - Efficient protein production using a Bombyx mori nuclear polyhedrosis virus lacking the cysteine proteinase gene. AB - Infection by a baculovirus (Bombyx mori nuclear polyhedrosis virus, BmNPV) in silkworm (Bombyx mori) larvae is highly efficient as an expression system for the production of useful proteins. However, the amount of the protein of interest expressed tends to decrease in the later stages of infection presumably due, in part, to a proteinase produced in the larval haemolymph. The N-terminal amino acid sequence of a proteinase purified from the haemolymph of BmNPV-infected larvae was identical to the internal amino acid sequence of the viral cysteine proteinase gene of BmNPV, suggesting that the cysteine proteinase in the haemolymph originated from the BmNPV gene. We constructed a mutant virus (CPd) which had a deletion in the cysteine proteinase gene. No proteinase activity corresponding to this proteinase was detected in the haemolymph of silkworm larvae infected with CPd. The firefly luciferase and the human growth hormone genes were separately introduced into CPd under control of the polyhedrin promoter. These constructs produced these proteins very efficiently, because of a greatly reduced degree of degradation of these proteins. A BmNPV vector system using CPd enhances the stability of foreign expressed proteins, especially for those that are cysteine proteinase-sensitive. PMID- 9400954 TI - Alcohol, tobacco and recreational drug use and the risk of non-Hodgkin's lymphoma. AB - A population based case-control study was conducted to determine whether risk of non-Hodgkin's lymphoma (NHL) in the absence of HIV infection is related to the previous use of tobacco, alcohol or recreational drugs. A total of 378 residents of Los Angeles County who were diagnosed with high- or intermediate-grade NHL were compared with individually age-, race- and sex-matched neighbourhood control subjects with regard to history of use of tobacco products, alcohol and ten specific recreational drugs. Risk of NHL among women decreased with increased consumption of alcoholic beverages (trend P = 0.03), with risk 50% lower among those consuming five or more drinks per week than among non-drinkers. Cocaine, amphetamines, Quaaludes and lysergic acid diethylamide (LSD) were each associated with a significantly increased risk of NHL in men with risk greater among those with more frequent use of these drugs. Confounding factors could not be excluded in these findings. The use of multiple types of drugs was also associated with a significantly increased risk of NHL in men (trend P = 0.005) with risk greatest among those using five or more types of drugs (odds ratio = 5.8, 95% confidence limits = 1.2-28.4); among these drugs, cocaine use appeared to account for the elevated risk of NHL among men based on multivariable analyses. PMID- 9400956 TI - Binding and fusion of Autographa californica nucleopolyhedrovirus to cultured insect cells. AB - Binding of baculoviruses to insect cells and fusion of the virus envelope to cell membranes are early events suggested to be affected by baculovirus enhancins. The binding of Autographa californica nucleopolyhedrovirus (AcMNPV) to the Spodoptera frugiperda cell line Sf21 and the fusion of the virus envelope to cell membranes were characterized. Virus binding assays demonstrated that AcMNPV budded virus (BV) bound to specific binding sites on Sf21 cells with an avidity of 2.3 x 10(10) M(-1). The cells displayed 3.1 x 10(3) specific binding sites per cell in a confluent monolayer. In addition, the effects of pH, buffer composition and cation concentration on the binding were examined. Using a fluorescent probe (R18) and fluorescence microscopy, the fusion of AcMNPV BV envelope to the cell membrane was directly visualized in living cells. It has been reported that Trichoplusia ni nucleopolyhedrovirus enters Sf21 cells by membrane fusion at the cell surface; however, the present studies confirmed the well established concept that adsorptive endocytosis is the major entry pathway for baculovirus BV infection. Membrane fusion kinetics and fluorescence microscopy demonstrated and verified that the envelope-cell membrane fusion was triggered by acidification. The effect of a T. ni granulovirus enhancin on virus binding and membrane fusion was examined, and no increase in activity was observed. PMID- 9400957 TI - Analysis of p74, a PDV envelope protein of Autographa californica nucleopolyhedrovirus required for occlusion body infectivity in vivo. AB - In nature, nuclear polyhedrosis viruses (NPV) are transmitted when susceptible insect larvae ingest viral occlusion bodies (OB). These dissociate in the alkaline environment of the midgut and release encapsulated virions (PDV) which bind to midgut epithelial cells and initiate an infection. A previous study showed that expression of the Autographa californica NPV (AcMNPV) p74 gene during replication is essential for the production of infectious OB. A set of p74 deletion and overexpression recombinants was used for the production and screening of monoclonal antibodies, and for an investigation of gross cytopathology and localization of p74. No differences in virus structure or morphogenesis were observed in infected cells when the p74 gene of AcMNPV was deleted, even though the infectivity of OB harvested from the cells was abolished when they were fed to Trichoplusia ni larvae. Mutant OB released virus particles and degraded insect peritrophic membrane as in infections with wild-type virus; in addition, virions purified from mutant OB were infectious when injected into the haemocoel of T. ni larvae. Western blot analysis confirmed that p74 was associated with the PDV and could not be detected in the budded form virion phenotype. The polypeptide was readily degraded by treatment of purified PDV with proteinase K, in the presence and absence of detergent, and could be extracted from PDV by a non-ionic detergent treatment. The data are consistent with p74 being a structural polypeptide of the PDV phenotype, most probably as a component associated with the outside surface of the virion envelope. Its presence is shown to be essential for primary infection of midgut cells of insect larvae. PMID- 9400958 TI - Characterization of a putative Spodoptera exigua multicapsid nucleopolyhedrovirus helicase gene. AB - Putative baculovirus helicases have been implicated as playing an important role in viral DNA replication and host specificity. The Spodoptera exigua multicapsid nucleopolyhedrovirus (SeMNPV) helicase is therefore of interest since the virus only infects the beet army worm. Sequence analysis of the SeMNPV lef5-p39 (mu 46.5-55.1) region, which is collinear with the 39K-lef5 area in Autographa californica MNPV (AcMNPV), revealed an open reading frame (ORF) of 3666 bp potentially encoding a protein with a molecular mass of 143 kDa. This protein had considerable amino acid sequence similarity (58%) to AcMNPV p143, including seven conserved motifs characteristic of helicases. In cultured insect cells, this SeMNPV ORF is expressed from 4 to 12 h postinfection and its major transcript of 4 kb starts 11 to 12 nt upstream of the putative translational initiation site (ATG). To study their possible role in the specificity of baculovirus DNA replication, the putative AcMNPV and SeMNPV helicase genes were tested for their ability to replicate homologous regions (hrs; putative origins of DNA replication) in a transient DNA replication assay in insect cells. All viral cis- and trans-acting factors were provided as plasmids using either Achr2 or Sehr1 as the DNA replication origin. SeMNPV p143 could not substitute for AcMNPV p143 in the transient assays supplemented with either hr. Similar results were obtained when the SeMNPV and AcMNPV ie1 genes were exchanged. None of the essential AcMNPV trans-acting factors could be complemented by SeMNPV infections to support DNA replication of hrs. These data suggest a specific interaction between baculovirus DNA replication factors to form the replisome and/or between the replisome and the origin of DNA replication. PMID- 9400959 TI - Mapping of the Heliothis armigera entomopoxvirus (HaEPV) genome, and analysis of genes encoding the HaEPV spheroidin and nucleoside triphosphate phosphohydrolase I proteins. AB - The genome of Heliothis armigera entomopoxvirus (HaEPV) has been mapped with four restriction endonuclease enzymes (BamHI, HindIII, PstI and XhoI), and its length estimated at 233 kbp. An EcoRI-generated HaEPV genomic fragment hybridized to all fragments identified as genomic termini, providing the first experimental evidence for the presence of terminal repeat elements in an EPV genome. The HaEPV spheroidin and nucleoside triphosphate phosphohydrolase I (NPHI) genes have been cloned and sequenced, and their deduced products shown to possess high levels of identity with homologues from other Genus B entomopoxviruses (EPVs). The genomic locations of these and other HaEPV genes and open reading frames have been determined; comparison of their locations with those of homologues in the Amsacta moorei EPV genome largely supports an hypothesis that the Genus B EPVs share a conserved genomic organization which differs from that of chordopoxviruses. It is proposed that genes of EPVs can be assigned to five actual or predicted homology based groups, a categorization which is useful for directing and interpreting investigations of EPV gene functions and relationships. PMID- 9400961 TI - Hypothesis on particle structure and assembly of rice dwarf phytoreovirus: interactions among multiple structural proteins. AB - To study the morphogenesis and packaging of rice dwarf phytoreovirus (RDV), the interactions among multiple structural proteins were analysed using both the yeast two-hybrid system and far-Western blotting analysis. The following protein protein interactions were observed. P3 (major core protein) bound to itself as well as to P7 (nucleic acid-binding protein) and P8 (major outer capsid protein). P7 bound to P1 (RNA-dependent RNA polymerase) and P8, in addition to P3. Based on these findings, we hypothesize that the core shell structure is based on P3-P3 interactions and that P7 has the ability to bind to multiple structural proteins as well as to genomic RNAs during viral particle assembly. Based on the observed protein-protein interactions and on computer-aided analysis of the numbers of structural proteins per particle, possible RDV assembly events are proposed. PMID- 9400960 TI - Excision of the polydnavirus chromosomal integrated EP1 sequence of the parasitoid wasp Cotesia congregata (Braconidae, Microgastinae) at potential recombinase binding sites. AB - Cotesia congregata polydnavirus (CcPDV) is essential for successful parasitism of Manduca sexta larvae by the braconid wasp Cotesia congregata. To determine the molecular mechanisms for the vertical transmission of CcPDV in the wasps, we analysed the different forms of the virus sequences containing the gene encoding the early parasitism-specific protein 1 (EP1). By a detailed molecular analysis, we demonstrated that the EP1 sequences are present in wasp DNA in two forms: a circular form as seen in the virus particles and a form integrated into the wasp genome. Moreover, we showed that the integrated form of the EP1 sequences is able to excise in the ovary cells. A fragment corresponding to an EP1 'empty locus' (without the viral sequence) was PCR-amplified from ovarian DNA. Comparison of the sequences isolated from the EP1 circle, the integrated form and the empty locus revealed that the extremities of the EP1 genomic sequences constitute a direct repeat. Strikingly, these sequences contain a potential binding site for a recombinase of the Hin family located in close vicinity to the position where the DNA strand exchange occurs. Thus, the data bear upon the possibility that the bracovirus circles are excised via a mechanism related to the Hin mediated Conservative Specific-Specific Recombination (CSSR) of prokaryotes. PMID- 9400962 TI - Infectious in vivo and in vitro transcripts from a full-length cDNA clone of PVY N605, a Swiss necrotic isolate of potato virus Y. AB - A full-length cDNA clone of the potato virus Y (PVY) genome was obtained after cloning difficulties in Escherichia coli were overcome. These difficulties were mainly due to the expression of the CI gene from upstream prokaryotic promoter like elements within the PVY genome. To overcome this problem, PVY cDNA was maintained in two subclones which were ligated before infection. A plasmid in which these two fragments were contained could be propagated in some E. coli strains but was unstable and yielded only low levels of plasmid DNA. Replacement of the 35S promoter by the SP6 promoter slightly increased the stability of the plasmid and its RNA transcripts were infectious when capped with m7G(5')ppp(5')G. Using two inoculation methods (mechanical or biolistic) the cDNA and its RNA transcript proved infectious on three Nicotiana species and on Solanum tuberosum. PMID- 9400963 TI - 35S promoter-driven cDNAs of barley mild mosaic virus RNA1 and RNA2 are infectious on barley plants. AB - Full-length cDNAs of barley mild mosaic bymovirus RNA1 and RNA2 were cloned downstream of a modified cauliflower mosaic virus 35S promoter with double enhancer. Mechanical inoculation of barley seedlings with a mixture of both cDNAs resulted in systemic mosaic symptoms, typical of barley mild mosaic virus infection. The presence of both RNA species and their gene products in the systemically infected leaves was demonstrated by RT-PCR and Western blot analyses, respectively. Virions were detected by immunogold labelling, demonstrating that the RNAs are encapsidated. This is the first report of the 35S promoter used in successfully infecting a monocot plant host with cDNA from a strictly monocot plant RNA virus. PMID- 9400964 TI - Variability of geographically distinct isolates of maize rayado fino virus in Latin America. AB - We have examined the molecular epidemiology of the leafhopper-borne maize rayado fino virus (MRFV) in Latin America. The coat protein gene and 3' non-translated region of 14 isolates of MRFV collected from Latin America and the United States were sequenced and phylogenetic relationships examined. The nucleotide sequence revealed remarkable conservation, with a sequence similarity of 88-99%. Phylogenetic analysis of sequence data obtained from a 633 bp fragment showed that MRFV has diverged into three main clusters, i.e. the geographically distinct northern and southern isolates and the Colombian isolates. Significant differences between the isolates collected from Colombia, previously named maize rayado colombiana virus, based upon differences in symptomatology and serological relationships to MRFV, and the other MRFV isolates, provides additional evidence supporting its designation as a unique strain of MRFV. PMID- 9400965 TI - Beet soil-borne virus RNA 1: genetic analysis enabled by a starting sequence generated with primers to highly conserved helicase-encoding domains. AB - The complete sequence of the 5834 nucleotides of RNA 1 of beet soil-borne furovirus (BSBV, Ahlum isolate) was determined using a PCR product obtained with primers to highly conserved coding regions for helicase-like proteins in RNA 1 of furo-, hordei- and tobraviruses as a starting sequence. Unknown parts of the sequence upstream and downstream of this starting sequence were amplified by means of RT-PCR techniques using combinations of specific and random primers. BSBV RNA 1 contains one large ORF for a readthrough protein with a molecular mass of 204 kDa (204K protein) which is interrupted internally by a UAA stop codon terminating the coding region for a protein of 145 kDa (145K protein). The N- and C-terminal parts of the 145K protein and the readthrough domain of the 204K protein contain methyltransferase, helicase and RNA-dependent RNA polymerase motifs, respectively. Unlike other furo- and tobraviruses BSBV contains no further genes on its RNA 1. PMID- 9400966 TI - Isolation and characterization of tubular structures of cowpea mosaic virus. AB - Tubular structures involved in the cell-to-cell movement of cowpea mosaic virus (CPMV) were partially purified from infected cowpea protoplasts to identify the structural components. A relatively pure fraction could be obtained by differential centrifugation and this was analysed by PAGE and immunoblotting. Besides the movement protein (MP) and capsid proteins (CP) of CPMV, no other major infection-specific proteins could be detected, suggesting that host proteins are not a major structural component of the movement tubule. PMID- 9400967 TI - Replication of alfalfa mosaic virus RNA 3 with movement and coat protein genes replaced by corresponding genes of Prunus necrotic ringspot ilarvirus. AB - Alfalfa mosaic virus (AMV) and Prunus necrotic ringspot virus (PNRSV) are tripartite positive-strand RNA plant viruses that encode functionally similar translation products. Although the two viruses are phylogenetically closely related, they infect a very different range of natural hosts. The coat protein (CP) gene, the movement protein (MP) gene or both genes in AMV RNA 3 were replaced by the corresponding genes of PNRSV. The chimeric viruses were tested for heterologous encapsidation, replication in protoplasts from plants transformed with AMV replicase genes P1 and P2 (P12 plants) and for cell-to-cell transport in P12 plants. The chimeric viruses exhibited basic competence for encapsidation and replication in P12 protoplasts and for a low level of cell-to cell movement in P12 plants. The potential involvement of the MP gene in determining host specificity in ilarviruses is discussed. PMID- 9400968 TI - Hop stunt viroid (HSVd) sequence variants from Prunus species: evidence for recombination between HSVd isolates. AB - Hop stunt viroid (HSVd) is able to infect a number of herbaceous and woody hosts, such as grapevine, Citrus or Prunus plants. Previous phylogenetic analyses have suggested the existence of three major groups of HSVd isolates (plum-type, hop type and citrus-type). The fact that these groups often contain isolates from only a limited number of isolation hosts prompted the suggestion that group discriminating sequence variations could, in fact, represent host-specific sequence determinants which may facilitate or be required for replication in a given host. In an effort to further understand the relationships between HSVd and its different hosts, HSVd variants from eight naturally infected Prunus sources, including apricot, peach and Japanese plum have been cloned and sequenced. In total, ten molecular variants of HSVd have been identified, nine of which have not been described before. A detailed phylogenetic analysis of the existing HSVd sequences, including the new ones from Prunus determined in this work, points towards a redefinition of the grouping of variants of this viroid, since two new groups were identified, one of them composed of sequences described here. A bias for the presence of certain sequences and/or structures in certain hosts was observed, although no conclusive host-determinants were found. Surprisingly, our analysis revealed that a number of HSVd isolates probably derived from recombination events and that the previous hop-type group itself is likely to be the result of a recombination between members of the plum-type and citrus-type groups. PMID- 9400969 TI - Characterization of genotype-specific epitopes of the HN protein of mumps virus. AB - The SBL-1 strain of mumps virus, grouping with genotype A on the basis of the small hydrophobic protein gene sequence, was grown in the presence of three different monoclonal antibodies. The monoclonal antibodies were directed against the haemagglutinin-neuraminidase (HN) protein and they inhibited haemagglutinating activity and infectivity of the virus. The HN genes of the nine neutralization-escape virus mutants were sequenced and the predicted amino acid sequences were compared with that of the parental virus. Amino acid substitutions were found at positions 269, 352 and 354, respectively, of the 582 amino acid long protein. The three monoclonal antibodies did not react with 35 virus strains isolated in Stockholm during the years 1970 to 1985. Thirteen and four of the strains were found to belong to the D and C genotypes, respectively. A type specific neutralization antibody response was also found in sera of rabbits hyperimmunized with purified virions of genotype A and D. The genotype-specific difference in neutralizing activity in mice and rabbits was not corroborated by an overall difference in the amino acid sequence of the HN protein of the different genotypes. Further studies are needed to explore the efficacy of mumps virus vaccines for protection against homologous and heterologous genotypes of mumps virus. PMID- 9400970 TI - Recombinant vaccinia viruses expressing the F, G or N, but not the M2, protein of bovine respiratory syncytial virus (BRSV) induce resistance to BRSV challenge in the calf and protect against the development of pneumonic lesions. AB - The immunogenicity and protective efficacy of recombinant vaccinia viruses (rVV) encoding the F, G, N or M2 (22K) proteins of bovine respiratory syncytial virus (BRSV) were evaluated in calves, the natural host for BRSV. Calves were vaccinated either by scarification or intratracheally with rVV and challenged 6 to 7 weeks later with BRSV. Although replication of rVV expressing the F protein in the respiratory tract was limited after intratracheal vaccination, the levels of serum and pulmonary antibody were similar to those induced following scarification. The serum antibody response induced by the F protein was biased in favour of IgG1 antibody, whereas the G and the N proteins induced similar levels of IgG1:IgG2, and antibody was undetectable in calves primed with the M2 protein. The F protein induced neutralizing antibodies, but only low levels of complement dependent neutralizing antibodies were induced by the G protein, and antibody induced by the N protein was not neutralizing. The F and N proteins primed calves for BRSV-specific lymphocyte proliferative responses, whereas proliferative responses were detected in calves primed with the G protein only after BRSV challenge. The M2 protein primed lymphocytes in only one out of five calves. Although there were differences in the immune responses induced by the rVVs, the F, G and N, but not the M2, proteins induced significant protection against BRSV infection and, in contrast with the enhanced lung pathology seen in mice vaccinated with rVV expressing individual proteins of human (H)RSV, there was a reduction in lung pathology in calves. PMID- 9400971 TI - Isolation of an avirulent mutant of Sendai virus with two amino acid mutations from a highly virulent field strain through adaptation to LLC-MK2 cells. AB - A field strain of Sendai virus (SeV) Ohita-M1 (M1) was isolated from an epidemic in an animal laboratory by passaging in mice. A mutant strain, Ohita-MVC11 (MVC11), was then obtained by passaging M1 in rhesus monkey (LLC-MK2) cells. MVC11 was adapted to LLC-MK2 cells and produced 20 times higher levels of infectious virus than M1. This increased production of infectious virus in LLC MK2 cells was associated with enhanced viral gene expression. However, MVC11 could not replicate efficiently in mouse lung and was not lethal to mice even when inoculated at a titre of 8 x 10(5) cell-infecting units (CIU) per mouse. On the other hand, with an inoculum of only 4 x 10(1) CIU per mouse, corresponding to 1 LD50, M1 replicated well in mouse lung and was highly virulent to mice. Nucleotide and deduced amino acid sequence analyses of the entire genomes of M1 and MVC11 revealed that adaptation to LLC-MK2 cells and the attenuation of mouse pathogenicity of MVC11 were associated with only two amino acid substitutions; one on the C protein (Phe substituted by Ser at position 170) and the other on the RNA polymerase, the L protein (Glu substituted by Ala at position 2050). PMID- 9400972 TI - Cell cycle arrest rather than apoptosis is associated with measles virus contact mediated immunosuppression in vitro. AB - Acute measles is associated with pronounced immunosuppression characterized both by leukopenia and impaired lymphocyte functions. In an earlier study, we found that mitogen-dependent proliferation of uninfected human peripheral blood lymphocytes (PBLs) and spontaneous proliferation of human cell lines of lymphocytic or monocytic origin was impaired after contact with UV-inactivated, measles virus (MV)-infected cells, UV-inactivated MV or with cells transfected with MV glycoproteins (gp) F and H. We now show that mitogen-stimulated PBLs and Jurkat cell clones either highly sensitive or resistant to CD95-induced apoptosis have a similar sensitivity to MV-induced inhibition and do not undergo apoptosis. Moreover, unimpaired mitogen-dependent upregulation of important activation markers, including IL-2R, was observed in PBL cultures after contact with MV infected, UV-irradiated presenter cells. This indicates that the cells were indeed viable and acquire a state of activation. Less IL-2 was released from PBLs after contact with MV-infected presenter cells when compared with that released after contact with uninfected cells. However, mitogen-induced proliferation of PBLs was not restored by addition of IL-2 under these conditions. It appeared that a higher fraction of mitogen-stimulated PBLs accumulated in the G0/G1 phase of the cell cycle after contact with MV-infected cells. Thus, the mitogen unresponsiveness of PBLs seen after contact with MV-infected cells is due to cell cycle arrest rather than apoptosis. PMID- 9400973 TI - Fine specificity of the antibody response to a synthetic peptide from the fusion protein and protection against measles virus-induced encephalitis in a mouse model. AB - A synthetic peptide representing residues 397-420 from the measles virus (MV) fusion (F) protein was tested for its structure, immunogenicity and protective capacity against intracerebral challenge with a neuroadapted strain of MV. Analysis of the peptide by mass spectrometry showed that it was linear, despite the presence of two cysteine residues in the sequence. Circular dichroism spectroscopy highlighted a weak preference for the peptide to adopt an alpha helical conformation. The peptide was shown to be immunogenic in BALB/c and C57BL/6 mice after intraperitoneal immunization in Freund's adjuvant, and anti peptide antibodies from both strains of mice reacted with the MV as a solid phase antigen on an ELISA plate. When the fine specificity of the anti-peptide antibody response was examined using overlapping 8-mer peptides, serum antibodies from BALB/c mice recognized the region between residues 407-417 whereas antibodies from C57BL/6 mice recognized the region 408-420 of the 397-420 peptide sequence. Although anti-397-420 antibodies had no demonstrable neutralizing activity, protection against challenge with a neuroadapted strain of MV was demonstrated following active immunization with peptide in C57BL/6 mice or after passive transfer of anti-peptide antibodies in BALB/c mice. These findings highlight the importance of the 397-420 region in the induction of protective antibodies in the MV encephalitis mouse model, and suggest that this epitope might be a good candidate for inclusion in a future MV synthetic peptide vaccine. PMID- 9400974 TI - Biosynthesis, intracellular transport and enzymatic activity of an avian influenza A virus neuraminidase: role of unpaired cysteines and individual oligosaccharides. AB - Intracellular transport, glycosylation, tetramerization and enzymatic activity of the neuraminidase (NA) of fowl plague virus (FPV) were analysed in vertebrate cells after expression from a vaccinia virus vector. Tetramerization occurred with a half-time of 15 min, whereas passage through the medial Golgi apparatus and transport to the plasma membrane occurred with half-times of 2 and 3 h, respectively, suggesting a step in NA maturation beyond tetramerization that limits the rate of transport to the medial Golgi. NA transport rates were about fourfold slower than those of haemagglutinin (HA). Slow transport and processing of FPV NA was not altered by coexpression of FPV HA, nor was the transport rate of HA influenced by NA. The slow transport kinetics of NA were also observed in FPV-infected CV-1 cells. As deduced from the coding sequence, FPV NA has the shortest stalk of all naturally occurring NAs described to date and contains only three potential N-glycosylation sites, which are all located in the globular head domain. Elimination of each of the three N-glycosylation sites revealed that the two oligosaccharides at positions 124 and 66 are of the complex type, whereas the one at Asn-213 remains in mannose-rich form. The glycosylation mutants showed also that oligosaccharides at positions 124 and 213 of FPV NA modulate enzymatic activity. Transport of NA is not influenced by single elimination of any of the three oligosaccharide attachment sites. Mutational analysis of the three Cys residues not involved in intrachain disulfide pairing revealed that Cys-49 in the stalk of the NA molecule is responsible for the formation of disulfide-linked dimers. Analysis of cysteine mutants of FPV NA also demonstrated that disulfide linked dimers are not absolutely necessary for the formation of enzymatically active tetramers but may stabilize the quaternary structure of NA. PMID- 9400975 TI - Identification of a single genotype of hepatitis G virus by comparison of one complete genome from a healthy carrier with eight from patients with hepatitis. AB - Different isolates of a putative hepatitis virus called hepatitis GB virus C or hepatitis G virus (HGV) have been cloned recently from patients with hepatitis. This virus has also been found commonly in healthy carriers. We have cloned and sequenced a complete HGV genome, designated HGVCN, from a healthy Chinese blood donor. HGVCN shares 85.8-90.0% nucleotide sequence identity and 95.4-97.5% amino acid identity with the eight available full-length HGV genomes. Furthermore, the majority (82.8%) of the nucleotide substitutions found in HGVCN were synonymous and a fairly uniform distribution of changes was found across the entire genome without identification of any hypervariable region. When compared with the African isolates GBVC and GBVC-EA, the HGVCN-encoded polyprotein contained a 31 amino acid N-terminal extension which was predicted to be a defective core-like sequence. The sequences of the HGV E1 and E2 proteins displayed unique motifs and were highly conserved. Phylogenetic analysis revealed that all nine complete HGV isolates were closely related and that HGVCN grouped with the other Chinese HGV isolate (HGVC964). Taken together, our findings suggest that there is one single genotype of HGV and that the HGV genome cloned from the healthy carrier is not significantly different from those derived from patient sera. PMID- 9400976 TI - Molecular analysis of the differential restriction of human immunodeficiency virus type 1 replication in neuronal cell lines. AB - Human immunodeficiency virus type 1 (HIV-1) replication is restricted partially in SK-N-MC and completely in SK-N-SH neuronal cells. To investigate the molecular mechanism of this differential restriction of HIV-1 replication, cells infected with HIV-1 were analysed for their steady-state levels of: total and unintegrated HIV-1 DNA by DNA PCR, different species of HIV-1 RNA by RT-PCR, and HIV-1 p24 protein production by an ELISA procedure. We found that the kinetics of the infection were slower and there was a lower level of accumulation of HIV-1 macromolecules (total and unintegrated circular DNA, unspliced and spliced RNAs and viral proteins) in the SK-N-MC cells than in the permissive CEM cells. In SK N-SH cells, HIV-1 DNA was only transiently detected during the first 24 h post infection, and the unspliced RNA was detected up to 1 week post-infection. However, the HIV-1 spliced RNAs and the 2-LTR circular DNA were not detected at all during the course of infection. Both SK-N-MC and SK-N-SH cells showed higher levels of HIV-1 DNA, RNA and p24 protein when infected with an HIV-1 (amphotropic retrovirus) pseudotype, HIV-1B. However, the level of HIV-1 replication was still lower in SK-N-SH than in SK-N-MC cells. Moreover, although the kinetics of viral protein production were comparable in SK-N-MC cells infected with HIV-1B and CEM cells infected with HIV-1, the overall level of virus replication was still much lower in HIV-1B-infected SK-N-MC cells. These data suggest that the restriction of HIV-1 replication in neuronal cell lines takes place at both virus-entry and post-entry levels, and cellular factors may be involved in the differential restriction of HIV-1 replication in these cells. PMID- 9400977 TI - Salmonella typhimurium aroA recombinants and immune-stimulating complexes as vaccine candidates for feline immunodeficiency virus. AB - Two experimental feline immunodeficiency virus (FIV) vaccines were tested, either alone or in combination, in four groups of cats (A-D). One vaccine (SL3261-FIV) was composed of live attenuated Salmonella typhimurium aroA (SL3261) strains expressing the capsid (Gag) and part of the envelope (Env) proteins of FIV. The other was composed of FIV Gag and Env proteins incorporated into immune stimulating complexes (iscom-FIV). Cats of group A were immunized four times with SL3261-FIV. Cats of group B were immunized twice with SL3261-FIV and then twice with iscom-FIV. Cats of group C were immunized twice with SL3261 expressing the B subunit of cholera toxin (SL3261-CtxB) and then twice with iscom-FIV. Cats of group D, which served as negative controls, were immunized twice with SL3261-CtxB and then twice with iscom into which the Gag and Env proteins of simian immunodeficiency virus (SIV) had been incorporated (iscom-SIV). Two weeks after the last immunization, all cats were challenged with FIV. At this time, cats immunized with iscom-FIV (groups B and C) showed strong plasma antibody responses to Gag and Env, whilst these responses were weak or undetectable in the cats immunized four times with SL3261-FIV (group A). Seven weeks after FIV challenge, Env-specific antibody responses had increased considerably in cats of all groups except group A. The mean virus loads in the cats of this group proved to be lower than those of the other groups at all time points, indicating partial protection. PMID- 9400979 TI - Regulatory interactions of transcription factor YY1 with control sequences of the E6 promoter of human papillomavirus type 8. AB - Human papillomavirus type 8 (HPV-8) is a strictly cutaneous oncogenic virus known to induce malignant skin lesions in epidermodysplasia verruciformis patients. Our study shows that sequences surrounding transcription start sites of the HPV-8 oncogene E6 (nt 175-179) and comprising the presumable CCAAC and TATA boxes of the E6 promoter P175 contain a cluster of four motifs similar to the DNA recognition site of the multifunctional cellular transcription factor yin-yang 1 (YY1). Using DNase I footprinting and gel retardation tests it could be demonstrated that three of these motifs indeed act as YY1 binding sites. To test their functional relevance for P175 activity, engineered YY1 binding site mutants were analysed in the context of a P175 test vector using transient expression assays with human keratinocytes. YY1 turned out to exert both positive and negative effects upon the activity of the HPV-8 E6 promoter; binding of YY1 to a site upstream of the promoter's cap-position (BS1) activated transcription, whereas the downstream site (BS2) mediated repression. The second downstream YY1 binding site (BS3) seemed to play an auxiliary role, enhancing the negative effect of YY1 BS2. These observations define YY1 as an important cellular protein involved in the control of E6 oncogene expression of the skin-specific 'high risk' HPV-8 and emphasize the potential regulatory role of sequences located downstream of the transcription start site. PMID- 9400978 TI - Role of the Fas/Fas ligand pathway in apoptotic cell death induced by the human T cell lymphotropic virus type I Tax transactivator. AB - Two distinct human diseases have been described in association with human T cell lymphotropic virus type I (HTLV-I) infection: adult T cell leukaemia and tropical spastic paraparesis/HTLV-I-associated myelopathy. Although comprehensive understanding of specific mechanisms underlying pathogenesis of either disease has not yet been achieved, the viral regulatory protein Tax is believed to play a significant role. Previous studies demonstrated the potential of Tax to transform host cells. Here, it is shown that the Tax transactivator has in addition the potential to induce T cell death by apoptosis. Using an inducible system (Jurkat cell line JPX-9), significant apoptotic cell death upon Tax expression was observed. In an attempt to detect the cellular genes mediating this effect, it was found that induction of Tax was associated with marked upregulation of the Fas ligand (FasL) gene. Tax-induced apoptosis was inhibited when the Fas/FasL pathway was interrupted by YVAD-cmk, the inhibitor of ICE-like proteases. Transient expression experiments provided additional support for the putative role of endogenous FasL in Tax-induced apoptosis. Upon cotransfection with Tax expressing plasmid, the transcriptional activity of the FasL promoter was found to be significantly upregulated in Jurkat cells and several other cell lines, as measured by reporter gene expression. Furthermore, cotransfection using different Tax mutants demonstrated that both CREB and NF-kappaB activation domains of Tax protein were required for the transactivation to take effect. PMID- 9400980 TI - Preferential virosomal location of underphosphorylated H5R protein synthesized in vaccinia virus-infected cells. AB - The phosphorylation state of vaccinia virus (VV) protein H5R synthesized in infected cells was investigated by two-dimensional gel electrophoresis. Most of the H5R protein was underphosphorylated (pI 5.9 to 6.8) and, on centrifugation of cell lysates, was associated with virosomes sedimenting with nuclei. However, about a quarter of the H5R protein synthesized was highly phosphorylated (pI 5.5), and this was the major form of the H5R protein present in cytoplasmic extracts. Immunofluorescence of VV-infected cells in the absence of DNA replication showed that underphosphorylated H5R protein, specifically recognized by antibody, was abundantly distributed throughout the cytoplasm but also present in punctate particles, whereas most of the B1R protein detected was in the punctate particles. Late gene expression was not required for the H5R protein to accumulate in virosomes--viral DNA synthesis was sufficient. The different phosphorylation states and cytological locations of the H5R protein suggest it has multiple roles in VV development. PMID- 9400981 TI - Effective priming of neonates born to immune dams against the immunogenic pseudorabies virus glycoprotein gD by replication-incompetent adenovirus-mediated gene transfer at birth. AB - One of the main limitations of the vaccination of neonates from vaccinated or infected mothers is the interference by inherited maternal antibodies, which are known to inhibit the immune response against both live and inactivated vaccines. The efficiency of bypassing this inhibition by the transfer of an immunogenic glycoprotein gene, the gD gene of pseudorabies virus (PRV), into neonates was explored. The experiments were conducted in 1-day-old piglets, which are immunocompetent at birth. The same transcription unit (gD of PRV under the control of the adenovirus major late promoter) was delivered intramuscularly at birth either in the form of naked DNA or cloned in the genome of a replication defective adenovirus. A booster injection of a conventional live PRV vaccine strain was given at 10 weeks of age, the replication of which was greatly restricted by the residual amounts of colostral antibodies in control animals. Piglets were challenged at the age of 16 weeks with a virulent PRV strain. The replication-defective adenovirus was able to efficiently prime piglets born to immune dams against gD in such a way that inoculation with the Bartha strain protected them against a subsequent challenge with the same level of efficacy in piglets born to naive or immune dams. In contrast, piglets born to immune dams into which the gD gene was not transferred, or transferred as naked DNA at birth, were not protected. These results open the way for early immunization of neonates born to vaccinated or infected mothers. PMID- 9400982 TI - Biologically safe, non-transmissible pseudorabies virus vector vaccine protects pigs against both Aujeszky's disease and classical swine fever. AB - Envelope glycoprotein D (gD) of pseudorabies virus (PRV) is essential for penetration but is not required for cell-to-cell spread. When animals are inoculated with a phenotypically complemented PRV gD mutant, the virus is able to spread locally by means of direct cell-to-cell transmission, but progeny virions released by infected cells are non-infectious because they lack gD. Therefore, the virus cannot be transmitted from inoculated animals to other animals. This property makes a PRV gD mutant an attractive candidate as a safe vaccine vector. To examine whether a self-restricted, non-transmissible PRV mutant can be used as a biologically safe vaccine vector, a gD/gE-negative PRV recombinant virus which expresses envelope glycoprotein E2 of classical swine fever virus was constructed. Vaccination of pigs showed that the recombinant virus was able to protect pigs against both Aujeszky's disease and classical swine fever. PMID- 9400983 TI - Cytokine production in the nervous system of mice during acute and latent infection with herpes simplex virus type 1. AB - Immunocytochemistry on serial paraffin sections was used to monitor the production dynamics of cytokines (IL-2, IL-4, IL-6, IL-10, IFN-gamma and TNF alpha) and viral antigens in the trigeminal ganglion (TG) and the central side of the dorsal root entry zone (DRE) of mice, following infection of the cornea with herpes simplex virus type 1. In normal TG, scattered satellite cells were TNF alpha+ and in the DRE, TNF-alpha+ and/or low numbers of IL-6+ cells were detected. On day 3 after infection, foci of TG neurons with viral antigens were surrounded by large numbers of TNF-alpha+ and/or IL-6+ cells and low numbers of IFN-gamma+ cells. IL-2+ and/or IL-4+ cells appeared later, when viral antigens had almost cleared. In the TG, the most striking changes occurred with TNF-alpha, with respect to its source (satellite cells, Schwann cells and infiltrating cells) and the extent and long duration of its production. TNF-alpha was the predominant cytokine throughout acute and latent infection and even by day 30, numbers of satellite cells expressing this cytokine were three times higher than those in normal ganglia. Moreover, in the DRE, TNF-alpha was the only cytokine detected during virus clearance and again, its production continued, along with that of IL-6, on days 20 to 30, in both infiltrating cells and astrocytes. Thus, cytokines, particularly TNF-alpha and perhaps IL-6, from infiltrating cells and resident glial cells may have a role both in virus clearance and in normal homeostatic mechanisms in the nervous system such as repair and protection of neurons from damage. PMID- 9400984 TI - Herpes simplex virus type 1 immediate early protein IE63 shuttles between nuclear compartments and the cytoplasm. AB - Herpes simplex virus type 1 (HSV-1) immediate early protein IE63, an essential nuclear protein, is pleiotropic in function and, at the post-transcriptional level, inhibits RNA splicing, interacts with cellular splicing small nuclear ribonucleoprotein particles (snRNPs), binds RNA and prevents the nucleocytoplasmic transport of intron-containing mRNAs. Here it is reported that IE63 is a nucleocytoplasmic shuttle protein able to travel from snRNP- and RNA rich nuclear foci to the cytoplasm, where it accumulates during actinomycin D treatment. This newly identified property suggests that IE63 facilitates nuclear export of HSV-1 transcripts, in addition to retaining intron-containing transcripts in the nucleus. The mechanism by which IE63 controls RNA export has yet to be defined. PMID- 9400985 TI - Herpes simplex virus 1716, an ICP 34.5 null mutant, is unable to replicate in CV 1 cells due to a translational block that can be overcome by coinfection with SV40. AB - Herpes simplex virus (HSV) mutants lacking the gene encoding infected cell protein (ICP) 34.5 exhibit an attenuated phenotype in models of pathogenesis and have been used for experimental cancer therapy. Recently it was shown that the HSV ICP 34.5 protein functions to prevent the host cell-induced double-stranded RNA-activated protein kinase (PKR)-dependent translational block that normally occurs during virus infection. We now report that an HSV ICP 34.5 mutant called HSV-1716 is unable to replicate in the simian kidney cell-derived line CV-1, due to a translational block. Moreover, we find that this block can be overcome by simian virus 40 (SV40). This has been shown directly by infecting CV-1 cells with SV40 and HSV-1716 simultaneously, and indirectly via HSV-1716 infection of COS-1 cells (CV-1 cells transformed by an origin-defective mutant of SV40 that codes for wild-type T antigen). The translational block is restored when infections are done in the presence of the phosphatase inhibitor okadaic acid. These results support, but do not directly prove, contentions that HSV ICP 34.5 interacts with the PKR pathway to restore translation in non-permissive cells, and that SV40 large T antigen has a similar functional role, but acts downstream of the site of ICP 34.5 interaction (eIF2alpha) in the pathway. Study of this CV-1/COS-1 system should allow further clarification of the virus-host interactions that underlie the restricted replication of HSV-1 ICP 34.5 gene null mutants. PMID- 9400987 TI - Development of a model for cytomegalovirus infection of oligodendrocytes. AB - The well-characterized human oligodendroglioma (HOG) cell line, cells of which resemble immature oligodendrocytes, was used to investigate the level of permissiveness to human cytomegalovirus (CMV) infection. Expression of CMV genes was incomplete following exposure of HOG cells to CMV, in contrast with results observed with the astroglioma cell line U373-MG (used as a positive control). However, treatment with phorbol 12-myristate 13-acetate (PMA) or with dibutryl cAMP (dbcAMP) plus the phosphatase inhibitor 1-isobutyl-3,3-methyl xanthine (IBMX) rendered the HOG cells fully permissive to CMV; down-regulation of HLA class I and production of virions were only observed under these conditions. In contrast to the findings seen with the HOG cell line, treatment of U373-MG cells with dbcAMP/IBMX or PMA did not interfere with CMV-induced down-regulation of HLA class I. However, these chemical stimulators reduced virus production in U373-MG cells by 30 and 70%, respectively. PMID- 9400986 TI - Analysis of cyclin-dependent kinase activity after herpes simplex virus type 2 infection. AB - Small DNA viruses (adenoviruses, simian virus 40, or human papillomaviruses) induce S-phase progression but prevent cell division to provide precursors for viral DNA replication. Herpes simplex viruses types 1 or 2 (HSV-1 or HSV-2) contain genes which encode DNA-metabolizing enzymes, for example, ribonucleotide reductase, thymidine kinase and dUTPase, suggesting that S-phase factors are not required for an efficient infection. However, several studies indicated that HSV induces some events that occur during cell-cycle progression. To determine if HSV 2 induces S-phase entry, we examined serum-arrested African green monkey kidney cells (CV-1) after infection. Two hours after infection steady-state levels of the S-phase-specific cyclin, cyclin A, increased. S-phase cyclin-dependent kinase activity (CDK2) was stimulated 10-fold 8 h after infection but decreased at 16 or 24 h after infection. Mitotic CDK activity (CDC2) was not activated after infection, in part due to decreases in CDC2 protein levels and inactivation of enzymatic activity resulting from tyrosine phosphorylation of CDC2. Furthermore, CDK4 activity was not dramatically affected by infection. These studies indicate that HSV-2 infection selectively activates CDK2 after infection but cell-cycle progression does not occur. We hypothesize that infection activates certain components of the cell cycle which enhance viral gene expression and DNA replication. PMID- 9400988 TI - PCR amplification is more sensitive than tissue culture methods for Epstein-Barr virus detection in clinical material. AB - In this study we have compared the use of PCR and conventional tissue culture methods to detect Epstein-Barr virus (EBV) in peripheral blood mononuclear cells and throat wash samples. The study population included 29 healthy adult and 20 immunocompromised EBV-seropositive donors. The results show significantly higher EBV detection rates by PCR than the tissue culture methods in throat wash samples from both donor groups (P < 0.01 in healthy donors and P < 0.009 in the immunocompromised donors) and in peripheral blood from the immunocompromised but not from the healthy donors (P < 0.008). Furthermore, when EBV DNA detection rates in throat wash cell pellet and supernatant fluid were compared, a higher positive result was obtained with the cell pellets which reached statistical significance in the immunocompromised group (P < 0.02). No correlation was found between positivity in throat wash and peripheral blood from the same donors. PMID- 9400989 TI - Expression of Epstein-Barr virus BMRF-2 and BDLF-3 genes in hairy leukoplakia. AB - The high level of Epstein-Barr virus (EBV) replication found in hairy leukoplakia (HL) provides a unique opportunity to study EBV expression in the oral epithelium. Screening of a cDNA library from an HL biopsy revealed expression of two genes not previously described in vivo: BMRF-2 and BDLF-3. Sequence analysis of the cDNAs demonstrated several nucleotide changes from the B95-8 sequence. In all six different HL strains studied, only one amino acid change was found in BMRF-2 relative to B95-8 and two amino acid changes were found in the BDLF-3 ORF. mRNA expression of both genes was localized to the lower prickle cell layer of the tongue epithelium. BMRF-2 protein expression was primarily detected in the cell nuclei of the upper prickle cell layer; immunoelectron microscopy revealed that BMRF-2 was associated with the nuclear chromatin. BDLF-3 protein expression was observed in the perinuclear space and cytoplasm of the prickle cells. BDLF-3 has recently been identified as a virion-associated protein, but the functions of BMRF-2 and BDLF-3 have not been elucidated. PMID- 9400990 TI - Involvement of cdc2-mediated phosphorylation in the cell cycle-dependent regulation of p185neu. AB - We previously reported cell cycle-dependent negative regulation of p185neu (decreased tyrosine phosphorylation and kinase activity, with electrophoretic mobility retarded by serine/threonine phosphorylation) in M phase and the escape of mutation-activated p185neu* from this regulation. Our present results showed that retardation of electrophoretic mobility occurs independently of the cells' transformed status. We found that normal p185neu lost its ability to dimerize in the M phase. We demonstrated a physical association between cdc2 (a serine/threonine kinase, active in M phase) and p185neu. We showed that the carboxy terminal portion of p185neu is phosphorylated in vitro by cdc2. Many phosphopeptides (at least three phosphoserine residues) unique to the M phase were identified, and the in vivo and in vitro phosphopeptide patterns were superimposable. In contrast, mutation-activated p185neu* dimerized in the M phase with no changes in electrophoretic mobility, failed to associate with cdc2 and no unique phosphoserine residues could be identified in the M phase (data not shown), consistent with the escape of p185neu* from cell cycle-dependent regulation. Our results suggest that this escape is an intrinsic property of the mutation-activated p185neu* independent of its ability to transform cells. Our results also suggest the involvement of serine/threonine kinases such as cdc2 in the cell cycle-dependent negative regulation of p185neu. PMID- 9400991 TI - Modular organization of the E2F1 activation domain and its interaction with general transcription factors TBP and TFIIH. AB - The transcriptional activator E2F1 regulates the expression of genes at the G1/S boundary. We have characterized interactions of the E2F1 activation domain with two general transcription factors, the TATA-box binding protein (TBP) and TFIIH. Two distinct binding sites on E2F1 were identified for TBP (amino acids 386-417 and 415-437) each of which supported activation in mammalian cells when expressed as a fusion to a heterologous DNA-binding domain. Neither of these minimal activation domains independently bound TFIIH; rather, the TFIIH binding site of E2F1 overlaps both domains. Loss of TFIIH-binding by E2F1 resulted in a 60-65% reduction in transactivation, suggesting that the E2F1/TFIIH interaction is important, but not essential, for transactivation. The retinoblastoma protein (Rb) binds directly to E2F1 and represses E2F1-mediated transactivation. We have demonstrated that recombinant Rb can compete with TBP and the p62 subunit of TFIIH for binding to immobilized E2F1. A tumorigenic form of Rb deficient in repressing E2F1-mediated transactivation is likewise deficient in displacing TBP from E2F1. We propose that competition between Rb and both TBP and TFIIH for binding to E2F1 is a mechanism by which Rb inhibits transactivation by E2F1. PMID- 9400992 TI - Influence of ATM function on telomere metabolism. AB - The ATM gene product, which is defective in the cancer-prone disorder ataxia telangiectasia, has been implicated in mitogenic signal transduction, chromosome condensation, meiotic recombination and cell cycle control. The ATM gene has homology with the TEL1 gene of yeast, mutations of which lead to shortened telomeres. To test the hypothesis that the ATM gene product is involved in telomere metabolism, we examined telomeric associations (TA), telomere length, and telomerase activity in human cells expressing either dominant-negative or complementing fragments of the ATM gene. The phenotype of RKO colorectal tumor cells expressing ATM fragments containing a leucine zipper (LZ) motif mimics that of ataxia telangiectasia (A-T) cells. These transfected RKO cells relative to transfected controls had a higher frequency of cells with TA and shortened telomeres, but no detectable change in telomerase activity. In addition, the percentage of cells with TA after gamma irradiation was higher in the transfected RKO cells with dominant negative activity of the ATM gene, compared to control cells. SV40 transformed fibroblasts derived from an A-T patient and transfected with a complementing carboxyl terminal kinase region of the ATM gene had a reduced frequency of cells with TA, with no effect on the telomere length or telomerase activity. The present studies using isogenic cells with manipulated ATM function demonstrate a role for the ATM gene product in telomere metabolism. PMID- 9400993 TI - High frequency of p53 mutations in human oral epithelial dysplasia and primary squamous cell carcinoma detected by yeast functional assay. AB - To determine the timing and actual incidence of p53 mutations in oral epithelial lesions, we examined 33 primary squamous cell carcinomas (SCCs), 14 dysplasias and six hyperplasias from Japanese patients by a combination of yeast functional assay and DNA sequencing. The assay detects mutations of p53 mRNA between codons 67 and 347 on the basis of the DNA-binding activity of the protein. Twenty-six SCCs (79%) and five dysplasias (36%) were positive for p53 mutation, while all six hyperplasias were negative for the mutation. Human papillomavirus type 16 E6 mRNA was detected in one of seven p53 mutation-negative SCCs by reverse transcription polymerase chain reaction (RT-PCR). We further examined p53 mutations in 17 Sri Lankan oral SCCs using the yeast functional assay and the single-strand conformation polymorphism analysis of PCR-amplified DNA fragments (PCR-SSCP) of exon 5-8. The mutations were confirmed by DNA sequencing and the detection sensitivity was compared between the two methods. Six samples (35%) were positive for p53 mutation in PCR-SSCP analysis, while nine samples (53%) were positive in yeast functional assay. This suggests that the incidence of p53 mutations has been considerably underestimated in the conventional SSCP analysis. The present data indicate that p53 mutations are extremely frequent in oral cancers in the Japanese, and suggest that the timing and significance of p53 mutation in oral tumor progression vary in different ethnic populations and areas. PMID- 9400994 TI - Identification and characterization of R-ras3: a novel member of the RAS gene family with a non-ubiquitous pattern of tissue distribution. AB - Members of the Ras subfamily of GTP-binding proteins, including Ras (H-, K-, and N-), TC21, and R-ras have been shown to display transforming activity, and activating lesions have been detected in human tumors. We have identified an additional member of the Ras gene family which shows significant sequence similarity to the human TC21 gene. This novel human ras-related gene, R-ras3, encodes for a protein of 209 amino acids, and shows approximately 60-75% sequence identity in the N-terminal catalytic domain with members of the Ras subfamily of GTP-binding proteins. An activating mutation corresponding to the leucine 61 oncogenic lesion of the ras oncogenes when introduced into R-ras3, activates its transforming potential. R-ras3 weakly stimulates the mitogen-activated protein kinase (MAPK) activity, but this effect is greatly potentiated by the co expression of c-raf-1. By the yeast two-hybrid system, R-ras3 interacts only weakly with known Ras effectors, such as Raf and RalGDS, but not with RglII. In addition, R-ras3 displays modest stimulatory effects on trans-activation from different nuclear response elements which bind transcription factors, such as SRF, ETS/TCF, Jun/Fos, and NF-kappaB/Rel. Interestingly, Northern blot analysis of total RNA isolated from various tissues revealed that the 3.8 kilobasepair (kb) transcript of R-ras3 is highly restricted to the brain and heart. The close evolutionary conservation between R-ras3 and Ras family members, in contrast to the significant differences in its biological activities and the pattern of tissue expression, raise the possibility that R-ras3 may control novel cellular functions previously not described for other GTP-binding proteins. PMID- 9400995 TI - Upregulation of the angiogenic factors PlGF, VEGF and their receptors (Flt-1, Flk 1/KDR) by TSH in cultured thyrocytes and in the thyroid gland of thiouracil-fed rats suggest a TSH-dependent paracrine mechanism for goiter hypervascularization. AB - Placenta growth factor (PlGF) and vascular endothelial growth factor (VEGF) represent two closely related angiogenic growth factors active as homodimers or heterodimers. Since goiters of the thyroid gland are extremely hypervascular, we investigated the expression of PlGF, VEGF and their receptors, Flt-1 and Flk 1/KDR, in a small panel of human goiters from patients with Graves's disease, in an animal model of thyroid goitrogenesis and in in vitro cultured thyroid cells. Here we report that the mRNA expression of PlGF, VEGF and their receptors is markedly enhanced in biopsies of goiters resected from Graves's patients. In vivo studies demonstrated that in the thyroid gland of thiouracil-fed rats, increased mRNA and protein expression of PIGF, VEGF, Flt-1 and Flk-1/KDR occurred subsequent to the rise in the serum thyroid stimulating hormone (TSH) levels and in parallel with thyroid capillary proliferation. In vitro studies confirmed the existence of such TSH-dependent paracrine communication between thyroid epithelial cells and endothelium since the conditioned medium collected from TSH stimulated thyrocytes acquired mitogenic activity for human umbilical vein endothelial (HUVE) cells. Altogether, these data suggest that PlGF and VEGF, released by thyrocytes in response to the chronic activation of the TSH receptor pathway, may act through a paracrine mechanism on thyroid endothelium. PMID- 9400996 TI - Wig-1, a new p53-induced gene encoding a zinc finger protein. AB - Wild type p53 expressed from a temperature-sensitive (ts p53) construct induces both G1 cell cycle arrest and apoptosis in the p53-negative J3D mouse T lymphoma line (Wang et al., 1995). Using differential display analysis, we have identified one new p53-induced gene, wig-1 (for wild type p53-induced gene 1), whose 7.6 kb and 2.2 kb transcripts are upregulated in ts p53-transfected J3D cells following induction of wild type p53 expression by temperature shift to 32 degrees C. The wig-1 transcripts were also induced in irradiated NIH3T3 and p21-/- fibroblasts but not in irradiated p53-/- fibroblasts. Whole body gamma irradiation caused induction of both wig-1 transcripts in mouse brain, testis, kidney, spleen and lung. A basal wig-1 expression was detected in brain, testis and kidney. The WIG 1 protein contains three zinc finger motifs and a putative nuclear localization signal. PMID- 9400997 TI - ErbB kinases and NDF signaling in human prostate cancer cells. AB - Prostate carcinoma (PCA) is the most commonly diagnosed malignancy in American men. Our knowledge of PCA growth regulation lags behind that of other cancers, such as breast and colon carcinomas. Among receptor tyrosine kinases, the ErbB family is most frequently implicated in neoplasia. We report here the expression of ErbB family kinases and their ligands in PCA cell lines and a xenograft. While ErbB1/EGFR, ErbB2/NEU, and ErbB3 were always observed in a distinct pattern, ErbB4 was not observed. Interestingly, while TGF-alpha was expressed in the majority of PCA lines, the ligand Neu Differentiation Factor/Heregulin (NDF) was expressed only in an immortalized, non-transformed prostate epithelial line. Concomitantly, there was a significant difference in biological response to these ligands. NDF inhibited LNCaP growth and induced an epithelial-like morphological change, in contrast to TGF-alpha, which accelerated cell growth. We also performed the first comprehensive analysis of NDF signaling in a prostate line. LNCaP stimulated with NDF demonstrated crosstalk between ErbB3 and ErbB2 which did not involve ErbB1. NDF also turned on several cascades, including those of PI3-K, ERK/MAPK, mHOG/p38 and JNK/SAPK, but not those of PLCgamma or the STAT family. This signaling pattern is distinct from that of TGF-alpha. The activation of mHOG by ErbB2 or ErbB3 has not been reported, and may contribute to the unusual phenotype. PI3-K activation is characterized by the formation of a striking 'activation complex' with multiple tyrosine-phosphorylated species, including ErbB3. Our studies provide a framework in which to dissect the growth and differentiation signals of prostate cancer cells. PMID- 9400998 TI - Induction of Mdm2 and enhancement of cell survival by bFGF. AB - Basic fibroblast growth factor (bFGF) can exert mitogenic and viability-promoting effects in a wide range of biological systems. The biochemical activities mediating the cell survival function of bFGF are largely unknown. We report here that exposure of fibroblasts to bFGF, which confers upon them increased survival, also causes at the same time an increase in cellular levels of the Mdm2 oncoprotein. Cells constitutively exposed to a bFGF autocrine loop are more refractory to killing by cisplatin. This increased chemoresistance coincides with elevated Mdm2 and reduced activation of the endogenous p53, resulting in inefficient transcriptional activation of the bax gene promoter. Importantly, unlike Mdm2 accumulation in fibroblasts exposed to DNA damage, induction of Mdm2 by bFGF does not occur through a p53-mediated pathway. The role of p53 in DNA damage-induced apoptosis and the ability of Mdm2 to block p53-mediated cell death are well established. These findings therefore suggest that induction of Mdm2 and the subsequent inhibition of p53 function may contribute, at least partially, to the anti-apoptotic effects of bFGF and possibly some other survival factors. PMID- 9400999 TI - Loss of heterozygosity on the long arm of human chromosome 7 in sporadic renal cell carcinomas. AB - Cytogenetic and molecular analysis of DNA sequences with highly polymorphic microsatellite markers have implicated allele loss in several chromosomal regions including 3p, 6p, 6q, 8p, 9p, 9q, 11p and 14q in the pathogenesis of sporadic renal cell carcinomas (RCCs). Deletions involving the long arm of chromosome 7 have not been described in RCCs although they have been seen in several other tumor types. However, there have been no detailed analysis of loss of heterozygosity (LOH) of 7q sequences in sporadic RCCs. We therefore studied LOH for DNA sequences on 7q with 10 highly polymorphic markers in 92 matched normal/tumor samples representing sporadic RCCs including papillary, nonpapillary, and oncocytomas in order to determine whether allelic loss could be detected in a tumor type with no visible 7q rearrangements at the cytogenetic level. We found chromosome 7q allele loss in 59 of 92 cases (64%) involving one, two, or more microsatellite markers. The most common allele loss included loci D7S522 (24%) and D7S649 (30%) at 7q31.1-31.2, a region that contains one of the common fragile sites, FRA7G. By comparative multiplex PCR analysis, we detected a homozygous deletion of one marker in the 7q 31.1-31.2 region in one tumor, RC21. These results support the idea that a tumor suppressor gene in 7q31 is involved in the pathogenesis of sporadic renal cell carcinomas. PMID- 9401000 TI - Pim1 cooperates with E2a-Pbx1 to facilitate the progression of thymic lymphomas in transgenic mice. AB - Mice transgenic for the leukemia oncogene E2A-PBX1 invariably develop lethal, high-grade T-cell lymphomas by 5 months of age. In this study, retroviral insertional mutagenesis was employed to identify oncogenes that cooperate with the E2A-PBX1 transgene in lymphomagenesis. Neonatal retroviral infection substantially reduced length of survival due to accelerated development of lymphomas (81 versus 130 days). The Pim1 gene was targeted by retroviral insertions in 48% of accelerated lymphomas whereas less than 5% contained activated c-Myc and none contained activated Pim2. However, Pim1 DNA rearrangements were frequently sub-stoichiometric and not present at all sites of involvement in an otherwise monoclonal lymphoma indicating that Pim1 activation occurred late in the course of lymphomagenesis. Tumor subpopulations containing activated Pim1 alleles displayed a substantial growth advantage over Pim1 negative cells following serial transfer to secondary, syngeneic recipients. Cooperative interactions were observed in intercrossed Pim1 and E2A-PBX1 transgenic mice in which all double transgenic progeny developed lethal, diffuse T lineage lymphomas by 3 months of age, whereas only 13% of E2A-PBX1 and none of Pim1 single transgenic intercross progeny developed lymphomas by 1 year. Tumors from double transgenic mice were monoclonal providing evidence that additional genetic events were required for transformation. Therefore, Pim1 and E2a-Pbx1 cooperate in T lineage lymphomagenesis but they are not sufficient and the role of Pim1 is more likely to be associated with tumor progression. PMID- 9401001 TI - Differences in the mechanisms of growth control in contact-inhibited and serum deprived human fibroblasts. AB - In the present work we studied mechanisms of growth control in contact-inhibited and serum-deprived human diploid fibroblasts. The observation that the effects on [3H]thymidine incorporation and reduction of retinoblastoma gene product phosphorylation were additive when contact-inhibition and serum-deprivation were combined led us to the conclusion that the underlying mechanisms might be different. Both contact-inhibition and serum-deprivation led to a strong decrease of cdk4-kinase-activity and cdk2-phosphorylation at Thr 160, while the total amounts of cdk4 and cdk2 remained constant. In contact-inhibited cells, we revealed a strong protein accumulation of the cdk2-inhibitor p27 and a slight, but significant increase of the cdk4-inhibitor p16. In serum-deprived cells, the protein levels in p27 and p16 remained low. In contrast, we detected a rapid decrease of cyclin D1 and cyclin D3 which did not occur in contact-inhibited cells. These results indicate that serum-deprivation and contact-inhibition have different mechanisms although they affect the same pathway cyclin D-cdk4, pRB, cyclin E-cdk2. PMID- 9401002 TI - Cell cycle re-entry following chemically-induced cell cycle synchronization leads to elevated p53 and p21 protein levels. AB - Mimosine (MIM) and aphidicolin (APH) are two agents frequently used in tissue culture-based experiments to achieve cell synchronization at late G1 and S phases. Following MIM or APH treatment of human cancer cell lines, a reversible growth arrest in late G1 and S phases of the cell cycle was correlated with moderate increases in p53 and p21 protein levels. Both p53-dependent and independent increases in p21 were observed following treatment with either agent. However, a striking increase in p21 protein levels and a continuous elevation in both p53 and p21 protein levels were observed over 48 h after cells re-entered the cell cycle following the chemically-induced synchronization. In addition, the increase in p21 protein levels typically seen following treatment of cells with DNA damaging agents, was enhanced when cells were treated with genotoxic agents following MIM or APH synchronization. These findings suggest that caution should be exercised when interpreting results from experiments using cell synchronization agents, in particular, studies designed to investigate p53- and p21-regulatory pathways. PMID- 9401003 TI - Fibrillin-1 mutations in Marfan syndrome and other type-1 fibrillinopathies. AB - Fibrillin is the major component of extracellular microfibrils and is widely distributed in connective tissue throughout the body. Mutations in the fibrillin 1 (FBN1) gene, on chromosome 15q21.1, have been found to cause Marfan syndrome, a dominantly inherited disorder characterised by clinically variable skeletal, ocular, and cardiovascular abnormalities. Fibrillin-1 mutations have also been found in several other related connective tissue disorders, such as severe neonatal Marfan syndrome, dominant ectopia lentis, familial ascending aortic aneurysm, isolated skeletal features of Marfan syndrome, and Shprintzen-Goldberg syndrome. Mutations are spread throughout the gene and, with the exception of neonatal Marfan syndrome, show no obvious clustering or phenotypic association. PMID- 9401004 TI - Molecular basis of heat labile hexosaminidase B among Jews and Arabs. AB - Genotyping individuals for Tay-Sachs disease (TSD) is mainly based on the heat lability of beta-hexosaminidase (Hex) A (alphabeta) and the heat stability of Hex B (betabeta). Mutations in the HEXB gene encoding the beta-subunits of Hex that result in heat-labile Hex B thus may lead to erroneous enzymatic genotyping regarding TSD. Utilizing single strand conformation polymorphism (SSCP) analysis for all 14 exons of HEXB followed by direct sequencing of aberrant fragments, we screened individuals whose Hex B was heat labile. A novel heat labile mutation, previously concluded to exist in the HEXB gene, was identified among Jews and Arabs as 1627 G-->A. One individual with heat labile Hex B (HLB) was negative for this novel mutation and for the known 1514 G-->A HLB mutation, proving that there exists at least one other unidentified HLB mutation. Based on these results, it is advisable to perform DNA tests for 1627 G-->A mutation in suspected HLB individuals. PMID- 9401005 TI - Analysis of mutations and chromosomal localisation of the gene encoding RFX5, a novel transcription factor affected in major histocompatibility complex class II deficiency. AB - MHC class II deficiency is a severe primary immunodeficiency characterised by the absence of major histocompatibility complex class II (MHC-II) gene expression. It is genetically heterogeneous and can result from defects in at least four different trans-acting regulatory genes required for transcription of MHC-II genes. One of these genes has recently been shown to encode a novel DNA binding protein called RFX5, which is one subunit of a heteromeric protein complex (RFX) that binds to the promoters of MHC-II genes. We have characterised the mutations in all four patients known to harbour a defect in the RFX5 gene and have mapped this new human disease gene to chromosome 1 band q21, a region frequently exhibiting chromosomal aberrations in a variety of preneoplastic and neoplastic diseases. PMID- 9401006 TI - Cystic fibrosis mutation frequencies in upstate New York. AB - Upstate New York patients (100) with cystic fibrosis (i.e., 200 CF chromosomes), 72 from the CF center in Syracuse and 28 from a Buffalo CF center, were analyzed for their CF-causing mutations using restriction enzyme digest, single-strand conformation analysis (SSCA), and Heteroduplex (HA) analysis. Polymerase chain reaction (PCR) amplified products from all 27 CFTR exons using primers that included flanking intron junction sequence were investigated. More than 120 known cystic fibrosis transmembrane conductance regulator (CFTR) disease-causing mutations were screened. Four novel CFTR disease-causing mutations were identified (N287Y in exon 6b, 1259insA in exon 8, R1070P in exon 17b, and CF?20kbdel14b-18). A detection rate of 96% of the combined Syracuse and Buffalo population CF chromosomes was obtained. PMID- 9401007 TI - Charcot-Marie-Tooth disease with intermediate motor nerve conduction velocities: characterization of 14 Cx32 mutations in 35 families. AB - Charcot-Marie-Tooth disease can be inherited either autosomal dominantly or recessively or linked to the X chromosome. X-linked dominant Charcot-Marie-Tooth disease (CMTX) is a sensorimotor peripheral neuropathy in which males have usually more severe clinical symptoms and decreased nerve conduction velocities than do females. CMTX is usually associated with mutations in exon 2 of the connexin 32 (Cx32) gene. DNA from 35 unrelated CMT patients, without the 17p11.2 duplication, but with median nerve conduction between 30 and 40 m/s, were tested for the presence of Cx32 mutations. The entire coding sequence of the Cx32 gene was explored using a rapid nonradioactive technique to detect single-strand conformation polymorphisms (SSCP) on large PCR fragments. Thirteen abnormal SSCP profiles were detected and characterized by sequencing. In addition, systematic sequencing of the entire Cx32 coding region in the remaining index cases revealed another mutation that was not detected by SSCP. A total of 14 mutations were found, five of which were not previously reported. These results demonstrate the high frequency (40%) of mutations in the coding region of the Cx32 gene in CMT patients with intermediate MNCV, without 17p11.2 duplications. Most of these mutations (93%) can be detected by SSCP. PMID- 9401008 TI - Two mutated HEXA alleles in a Druze patient with late-infantile Tay-Sachs disease. AB - Two affected HEXA alleles were found in an Israeli Druze Tay-Sachs child born to first-cousin parents. His paternal allele contained two adjacent changes in exon 5: delta496C, which resulted in a frameshift and premature termination codon 96 nucleotides downstream, and 498C-->G, a silent mutation. The maternal allele had a 835T-->C transition in exon 8 (S279P). Phosphoimaging quantitation of the parents' RNAs showed that the steady-state levels of mRNAs of the mutant exons 5 and 8 were 5% and 50%, respectively, of normal levels. The exon 5 mutated allele with the premature translation termination resulted in severe deficiency of Hex A. Transient expression of the exon 8 mutated alpha-chain cDNA in COS-1 cells resulted in deficiency of enzymatic activity. The child exhibited a late infantile-type disease. PMID- 9401009 TI - Analysis of (CAG)n size heterogeneity in somatic and sperm cell DNA from intermediate and expanded Huntington disease gene carriers. AB - The length of the CAG repeat responsible for Huntington disease has been analysed by two PCR methods in blood and sperm DNA of 13 expansion carriers, two carriers of intermediate alleles, and four normal subjects. The two methods consistently confirmed size heterogeneity, more pronounced in sperm and confined to the CAG stretch. Based on densitometric scanning of films, four indexes addressed to different features of the PCR pattern were used to quantitate mosaicism. These revealed strong correlations with CAG size and intergenerational instability. However, mosaicism did not show a greater similarity in sibs who shared the same HD chromosome, nor was correlated with instability in the proband's pedigree. Our data do not support the hypothesis that cis-acting factors play a major role in the instability and leave the CAG size per se as the major determinant of sperm cell CAG instability. PMID- 9401010 TI - Familial lipoprotein lipase (LPL) deficiency: a catalogue of LPL gene mutations identified in 20 patients from the UK, Sweden, and Italy. AB - The aim of this study was to identify mutations in the lipoprotein lipase (LPL) gene in 20 unrelated patients with familial lipoprotein deficiency (FLLD) and to investigate the genotype/phenotype relationship. The previously reported G188E mutation (Monsalve et al., J Clin Invest 86:728-734, 1990) was screened for and found to be present in seven individuals (12/40 alleles). In addition, three patients were heterozygous for the 2.0 kb insertion (Langlois et al., Proc Nalt Acad Sci US 86:948-952, 1989). Two approaches were taken for new mutation detection; single-strand conformation polymorphism and sequencing to identify micro-mutations in the proximal promoter and exons 1-9 of the LPL gene and Southern blotting to identify gross mutations. Ten different point mutations were found (W86G, A158T, H183Q, G188E, S193R, P207L, L252X, N291S, M301T, L303P). Additionally, a two nucleotide deletion in exon 6 (delta1006-1007), a six nucleotide deletion in exon 8 (delta1441-1447), and a silent substitution in the wobble position of codon E118 were identified. In vitro mutagenesis and expression in COS-B cells suggested that the A158T and S193R substitutions virtually abolished enzyme activity. In analysing the genotype/phenotype relationship, there was no strong association between age at diagnosis, severity of symptoms, lipid levels, and the nature/position of the mutation. Triglyceride levels, however, were higher in compound heterozygotes compared to true homozygotes, possibly reflecting increased instability of heterodimers. Overall, 29 of 40 (72.5%) mutant alleles were identified. Failure to identify the mutation in 11 alleles might reflect the inadequacy of the method or the possibility that mutations lie within regions of the gene not screened in the study because of lack of availability of sequence. PMID- 9401011 TI - Mutation of hMSH3 and hMSH6 mismatch repair genes in genetically unstable human colorectal and gastric carcinomas. AB - Mutations within microsatellite sequences, consisting of additions or deletions of repeat units, are known as the replication/repair error positive (RER+) phenotype or micorsatellite instability (MI). Microsatellite instability has been demonstrated in hereditary and sporadic colorectal carcinomas and is usually observed in noncoding regions of genomic DNA. However, relatively few coding region targets of MI have been identified thus far. Using PCR, we amplified regions encompassing (A)8 and (C)8 microsatellite tracts within hMSH3 and hMSH6 from 31 RER+ sporadic colorectal tumors, 8 hereditary colon cancers, 23 RER+ gastric carcinomas, and 32 RER- gastric tumors. Mutations were found in 11 (36%) of 31 sporadic colon carcinomas, 4 (50%) of 8 hereditary colorectal cancers, and 5 (22%) of 23 RER+ gastric carcinomas, but in only 2 (6%) of 32 RER- gastric carcinomas. These frameshift mutations cause premature stop codons downstream that are predicted to abolish normal protein function. Our results and those of others suggest that DNA mismatch repair genes, such as hMSH3 and hMSH6, are targets for the mutagenic activity of upstream mismatch repair gene mutations and that this enhanced genomic instability may accelerate the accumulation of mutations in RER+ tumors. PMID- 9401012 TI - Identification of 16 sulfamidase gene mutations including the common R74C in patients with mucopolysaccharidosis type IIIA (Sanfilippo A). AB - Mucopolysaccharidosis type IIIA (MPS IIIA or Sanfilippo A disease) is a storage disorder caused by deficiency of the lysosomal enzyme sulfamidase. Mutation screening, using SSCP/heteroduplex analyses on cDNA and genomic DNA fragments, was performed in a group of 42 European patients. Sixteen of the 17 different gene mutations characterized have not been previously described. The spectrum of gene lesions consists of two 1-bp deletions (1091delC, 1093delG), an 18-bp duplication (421ins18), a splice site mutation (IVS2-2A-->G), and 13 different missense point mutations. As in other lysosomal storage disorders, the phenotypic heterogeneity is associated with a considerable genetic heterogeneity. The missense mutation R74C, which alters an evolutionary conserved amino acid in the active site of the enzyme, was found on 56% of alleles of 16 Polish patients, whereas it was less frequent among German patients (21% of disease alleles). R245H, a previously reported common mutation, represents 35% of disease alleles in German patients, but only 3% in Polish patients. As the combined frequency of the common mutations (R74C and R245H) in German and Polish populations exceeds 55%, screening for these two mutations will assist molecular genetic diagnosis of MPS IIIA and allow heterozygote testing in these populations. PMID- 9401013 TI - The repeat expansion detection method in the analysis of diseases with CAG/CTG repeat expansion: usefulness and limitations. AB - The repeat expansion detection (RED) method was described to detect expansions of trinucleotide repeats of unknown chromosomal location. We have improved the RED method by the use of 8-mer oligonucleotides and assessed its usefulness in 30 samples from patients with spinocerebellar ataxia type 1 (SCA1), Huntington's disease (HD), and Machado Joseph's disease (MJD), for which the number of CAG/CTG repeats was determined by sequencing. There was a good correlation between the number of repeats detected by sequencing and those identified by RED. However, in 17% of samples, the RED gave additional fragments for ligation products of different size than the CAG/CTG repeat expansion detected in the sample by sequencing. The same was observed in a group of control subjects (n = 78) without known clinical abnormalities in which products of more than 40 repeats were detected in 27% of them, indicating that CAG/CTG repeat expansions are common in the general population. Wether this corresponds to unidentified loci with expansions deserves further investigation. PMID- 9401014 TI - Method for in situ investigation of mitochondrial DNA deletions. AB - A number of mitochondrial DNA (mtDNA) deletions have been recently identified in the tissues of patients with mitochondrial diseases and in elderly individuals. To investigate the distribution of mutant mitochondrial genomes within any particular tissue, we have developed a sensitive method based on indirect in situ PCR. Our experiments have shown that the new method had the advantage of selectively amplifying only mtDNA bearing the 4,977 bp deletion. We show that this method is more sensitive than in situ hybridization for detecting the 4977 bp mtDNA deletion while using only a low number of PCR cycles that minimize damage to tissue architecture. By using this method, we have demonstrated that the mutation does not occur uniformly among the cells of a given tissue/organ. This technique will be useful studying the distribution/localization of mtDNA mutations in individual cells of tissues and when combined with enzyme histochemical procedures in adjacent sections will enable the correlation between mtDNA mutations and bioenergy defects in single cells. PMID- 9401015 TI - An improved methodology for the detection of the common mutation in the FGFR3 gene responsible for achondroplasia. AB - Homozygous achondroplasia is a neonatal lethal condition which can only be diagnosed in the first trimester of pregnancy by molecular analysis. The vast majority of patients with achondroplasia have a G-->A substitution at position 1138 of the fibroblast growth factor receptor (FGFR3) cDNA sequence, resulting in the substitution of an arginine for a glycine residue at position 380 of the FGFR3 protein. This mutation has typically been detected by SfcI digestion of amplified genomic DNA. We have demonstrated that the SfcI digestion protocol does not consistently distinguish between DNA samples heterozygous and homozygous for the G1138A substitution, and illustrates how the misdiagnosis of a homozygous affected fetus for one carrying only one copy of the G1138A mutation could occur. We report here an improved, simple nonradioactive technique which can reliably and consistently detect the presence of the G1138A mutation both in the heterozygous and homozygous state. PMID- 9401016 TI - Biosynthesis of phosphatidic acid in lipid particles and endoplasmic reticulum of Saccharomyces cerevisiae. AB - Lipid particles of the yeast Saccharomyces cerevisiae harbor two enzymes that stepwise acylate glycerol-3-phosphate to phosphatidic acid, a key intermediate in lipid biosynthesis. In lipid particles of the s1c1 disruptant YMN5 (M. M. Nagiec et al., J. Biol. Chem. 268:22156-22163, 1993) acylation stops after the first step, resulting in the accumulation of lysophosphatidic acid. Two-dimensional gel electrophoresis confirmed that S1c1p is a component of lipid particles. Lipid particles of a second mutant strain, TTA1 (T. S. Tillman and R. M. Bell, J. Biol. Chem. 261:9144-9149, 1986), which harbors a point mutation in the GAT gene, are essentially devoid of glycerol-3-phosphate acyltransferase activity in vitro. Synthesis of phosphatidic acid is reconstituted by combining lipid particles from YMN5 and TTA1. These results indicate that two distinct enzymes are necessary for phosphatidic acid synthesis in lipid particles: the first step, acylation of glycerol-3-phosphate, is catalyzed by a putative Gat1p; the second step, acylation of lysophosphatidic acid, requires S1c1p. Surprisingly, YMN5 and TTA1 mutants grow like the corresponding wild types because the endoplasmic reticulum of both mutants has the capacity to form a reduced but significant amount of phosphatidic acid. As a consequence, an s1c1 gat1 double mutant is also viable. Lipid particles from this double mutant fail completely to acylate glycerol-3 phosphate, whereas endoplasmic reticulum membranes harbor residual enzyme activities to synthesize phosphatidic acid. Thus, yeast contains at least two independent systems of phosphatidic acid biosynthesis. PMID- 9401018 TI - Nucleoid structure and distribution in thermophilic Archaea. AB - Nucleoid structure and distribution in thermophilic organisms from the Archaea domain were studied. Combined phase-contrast and fluorescence microscopy of DAPI (4',6-diamidino-2-phenylindole)-stained Sulfolobus acidocaldarius and Sulfolobus solfataricus cells revealed that the nucleoids were highly structured. Different nucleoid distribution within the cells, representing different partition stages, was observed. The conformation of the nucleoids differed between exponentially growing and stationary-phase cells. Also, the stationary-phase cells contained two chromosomes, and the nucleoids occupied a larger part of the interior of the cells than in the exponentially growing cells. The part of the cell cycle during which fully separated nucleoids could be detected was short. Since the postreplication period is long in these organisms, there was a considerable time interval between termination of chromosome replication and completion of nucleoid separation, similar to the G2 phase in eukaryotic cells. The length of the visible cell constriction period was found to be in the same range as that of eubacteria. Finally, cell-cell connections were observed under certain conditions. Possible eubacterial, eukaryotic, and unique features of nucleoid processing and cell division in thermophilic archaea are discussed. PMID- 9401019 TI - The D-allose operon of Escherichia coli K-12. AB - Escherichia coli K-12 can utilize D-allose, an all-cis hexose, as a sole carbon source. The operon responsible for D-allose metabolism was localized at 92.8 min of the E. coli linkage map. It consists of six genes, alsRBACEK, which are inducible by D-allose and are under the control of the repressor gene alsR. This operon is also subject to catabolite repression. Three genes, alsB, alsA, and alsC, appear to be necessary for transport of D-allose. D-Allose-binding protein, encoded by alsB, is a periplasmic protein that has an affinity for D-allose, with a Kd of 0.33 microM. As was found for other binding-protein-mediated ABC transporters, the allose transport system includes an ATP-binding component (AlsA) and a transmembrane protein (AlsC). It was found that AlsE (a putative D allulose-6-phosphate 3-epimerase), but not AlsK (a putative D-allose kinase), is necessary for allose metabolism. During this study, we observed that the D-allose transporter is partially responsible for the low-affinity transport of D-ribose and that strain W3110, an E. coli prototroph, has a defect in the transport of D allose mediated by the allose permease. PMID- 9401017 TI - Characterization of genes encoding dimethyl sulfoxide reductase of Rhodobacter sphaeroides 2.4.1T: an essential metabolic gene function encoded on chromosome II. AB - Rhodobacter sphaeroides 2.4.1T is a purple nonsulfur facultative phototrophic bacterium which exhibits remarkable metabolic diversity as well as genomic complexity. Under anoxic conditions, in the absence of light and the presence of dimethyl sulfoxide (DMSO) or trimethylamine N-oxide (TMAO), R. sphaeroides 2.4.1T utilizes DMSO or TMAO as the terminal electron acceptor for anaerobic respiration, which is mediated by the molybdoenzyme DMSO reductase. Sequencing of a 13-kb region of chromosome II revealed the presence of 10 putative open reading frames, of which 5 possess homology to genes encoding the TMAO reductase (the tor system) of Escherichia coli. The dorS and dorR genes encode a sensor-regulator pair of the two-component sensory transduction protein family, homologous to the torS and torR gene products. The dorC gene was shown to encode a 44-kDa DMSO inducible c-type cytochrome. The dorB gene encodes a membrane protein of unknown function homologous to the torD gene product. The dorA gene encodes DMSO reductase, containing the molybdopterin active site. Mutations were constructed in each of these dor genes, and the resulting mutants were shown to be impaired for DMSO-dependent anaerobic growth in the dark. The mutant strains exhibited negligible levels of DMSO reductase activity compared to the wild-type strain under similar growth conditions. Further, no DorA protein was detected in DorS and DorR mutant strains with anti-DorA antisera, suggesting that the products of these genes are required for the positive regulation of dor expression in response to DMSO. This characterization of the dor gene cluster is the first evidence that genes of chromosome CII encode metabolic functions which are essential under particular growth conditions. PMID- 9401020 TI - Temperature-sensitive lesions in the Francisella novicida valA gene cloned into an Escherichia coli msbA lpxK mutant affecting deoxycholate resistance and lipopolysaccharide assembly at the restrictive temperature. AB - The valAB locus of Francisella novicida has previously been found to be highly similar at the deduced amino acid level to msbA lpxK of Escherichia coli. Both ValA and MsbA are members of the superfamily of ABC transporters, and they appear to have similar functions. In this study we describe the isolation of a temperature-sensitive valAB locus. DNA sequence analysis indicates that the only changes to the ValAB deduced amino acid sequence are changes of S453 to an F and T458 to an I in ValA. E. coli strains defective in msbA and expressing temperature-sensitive ValA rapidly ceased growth when shifted from a permissive temperature to a restrictive temperature. After 1 h at the restrictive temperature, cells were much more sensitive to deoxycholate treatment. To test the hypothesis that ValA is responsible for the transport or assembly of lipopolysaccharide, we introduced gseA, a Kdo (3-deoxy-D-manno-octulosonic acid) transferase from Chlamydia trachomatis, into a strain with a temperature sensitive valA allele and a nonfunctional msbA locus. These recombinants were defective in cell surface expression of the chlamydial genus-specific epitope within 15 min of a shift to the nonpermissive temperature. Also, there was enhanced association of the epitope with the inner membrane after a shift to the nonpermissive temperature. Thus, we propose that ValA is involved in the transport of lipopolysaccharide to the outer membrane. PMID- 9401021 TI - The minimal transactivation region of Saccharomyces cerevisiae Gln3p is localized to 13 amino acids. AB - Regulated nitrogen catabolic gene transcription in Saccharomyces cerevisiae is mediated by four positive (Gln3p and Gat1p/Nil1p) and negative (Dal80p/Uga43p and Deh1p/Nil2p/GZF3p) regulators which function in opposition to one another. All four proteins contain GATA-type zinc finger domains, and three of them (Gln3p, Dal80p, and Deh1p) have been shown to bind to GATA sequences situated upstream of genes whose expression is sensitive to nitrogen catabolite repression (NCR). The positive regulators, Gln3p and Gat1p, are able to support transcriptional activation when tethered by LexAp to the promoter of a reporter gene whose upstream activation sequences have been replaced with one or more lexA operator sites. Existing data suggest that these four proteins regulate transcription by competing with one another for binding to the GATA sequences which mediate NCR sensitive gene expression. We show that the minimal Gln3p domain mediating transcriptional activation consists of 13 amino acids with a predicted propensity to form an alpha-helix. Genetic analysis of this region (Gln3p residues 126 to 138, QQNGEIAQLWDFN) demonstrated that alanine may be substituted for the aromatic and acidic amino acids without destroying transcriptional activation potential. Similar substitution of alanine for the two hydrophobic amino acids, isoleucine and leucine, however, destroys activation, as does introduction of basic amino acids in place of the acidic residues or introduction of proline into the center of the sequence. A point mutation in the Gln3p activation region destroys its in vivo ability to support NCR-sensitive DAL5 expression. We find no convincing evidence that NCR regulates Gln3p function by modulating the functioning of its activation region. PMID- 9401023 TI - N-acyl-homoserine lactone-mediated regulation of phenazine gene expression by Pseudomonas aureofaciens 30-84 in the wheat rhizosphere. AB - Pseudomonas aureofaciens 30-84 is a soilborne bacterium that colonizes the wheat rhizosphere. This strain produces three phenazine antibiotics which suppress take all disease of wheat by inhibition of the causative agent Gaeumannomyces graminis var. tritici. Phenazines also enhance survival of 30-84 within the wheat rhizosphere in competition with other organisms. Expression of the phenazine biosynthetic operon is controlled by the phzR/phzI N-acyl-homoserine lactone (AHL) response system (L. S. Pierson III et al., J. Bacterial 176:3966-3974, 1994; D. W. Wood and L. S. Pierson III, Gene 168:49-53, 1996). By using high pressure liquid chromatography coupled with high-resolution mass spectrometry, the AHL produced by PhzI has now been identified as N-hexanoyl-homoserine lactone (HHL). In addition, the ability of HHL to serve as an interpopulation signal molecule in the wheat rhizosphere has been examined by using isogenic reporter strains. Disruption of phzI reduced expression of the phenazine biosynthetic operon 1,000-fold in the wheat rhizosphere. Coinoculation of an isogenic strain which produced the endogenous HHL signal restored phenazine gene expression in the phzI mutant to wild-type levels in situ. These results demonstrate that HHL is required for phenazine expression in situ and is an effective interpopulation signal molecule in the wheat rhizosphere. PMID- 9401022 TI - cps1+, a Schizosaccharomyces pombe gene homolog of Saccharomyces cerevisiae FKS genes whose mutation confers hypersensitivity to cyclosporin A and papulacandin B. AB - The Schizosaccharomyces pombe cps1-12 (for chlorpropham supersensitive) mutant strain was originally isolated as hypersensitive to the spindle poison isopropyl N-3-chlorophenyl carbamate (chlorpropham) (J. Ishiguro and Y. Uhara, Jpn. J. Genet. 67:97-109, 1992). We have found that the cps1-12 mutation also confers (i) hypersensitivity to the immunosuppressant cyclosporin A (CsA), (ii) hypersensitivity to the drug papulacandin B, which specifically inhibits 1,3-beta D-glucan synthesis both in vivo and in vitro, and (iii) thermosensitive growth at 37 degrees C. Under any of these restrictive treatments, cells swell up and finally lyse. With an osmotic stabilizer, cells do not lyse, but at 37 degrees C they become multiseptated and multibranched. The cps1-12 mutant, grown at a restrictive temperature, showed an increase in sensitivity to lysis by enzymatic cell wall degradation, in in vitro 1,3-beta-D-glucan synthase activity (173% in the absence of GTP in the reaction), and in cell wall biosynthesis (130% of the wild-type amount). Addition of Ca2+ suppresses hypersensitivity to papulacandin B and septation and branching phenotypes. All of these data suggest a relationship between the cps1+ gene and cell wall synthesis. A DNA fragment containing the cps1+ gene was cloned, and sequence analysis indicated that it encodes a predicted membrane protein of 1,729 amino acids with 15 to 16 transmembrane domains. S. pombe cps1p has overall 55% sequence identity with Fks1p or Fks2p, proposed to be catalytic or associated subunits of Saccharomyces cerevisiae 1,3 beta-D-glucan synthase. Thus, the cps1+ product might be a catalytic or an associated copurifying subunit of the fission yeast 1,3-beta-D-glucan synthase that plays an essential role in cell wall synthesis. PMID- 9401024 TI - Involvement of CysB and Cbl regulatory proteins in expression of the tauABCD operon and other sulfate starvation-inducible genes in Escherichia coli. AB - Starvation for sulfate results in increased synthesis of several proteins in Escherichia coli. Among these Ssi (sulfate starvation-induced) proteins are the products of the tauABCD genes, which are required for utilization of taurine as sulfur source for growth. In this study, the role of the cbl gene in expression of tauABCD and other ssi genes was investigated. The protein encoded by cbl shows high sequence similarity to CysB, the LysR-type transcriptional activator of the genes involved in cysteine biosynthesis. Strain EC2541, which contains an internal deletion in cbl, was unable to utilize taurine and other aliphatic sulfonates as sulfur sources. Two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis showed that many of the Ssi proteins were not synthesized in EC2541. Expression of a translational tauD'-'lacZ fusion required the presence of both cbl and cysB. The interactions of CysB and Cbl with the promoter region of tauABCD were studied by using gel mobility shift experiments and DNase I footprinting. CysB occupied multiple binding sites, whereas Cbl occupied only one site from 112 to 68 bp upstream of the transcription start site. Acetylserine, the inducer of transcription of CysB-regulated genes, stimulated binding of CysB but not of Cbl. Sulfate had no effect on binding of both proteins to the tauABCD promoter region. These results indicate that Cbl is a transcription factor for genes required for sulfonate-sulfur utilization and maybe for other genes whose expression is induced by sulfate starvation. PMID- 9401025 TI - Characterization of the aes gene of Escherichia coli encoding an enzyme with esterase activity. AB - malQ mutants of Escherichia coli lacking amylomaltase cannot grow on maltose. They express the maltose system constitutively and are sensitive to maltose when grown on another carbon source. In an attempt to isolate a multicopy suppressor that would result in growth on maltose, we transformed a malQ mutant with a gene bank of E. coli DNA which had been digested with Sau3a and cloned in pBR322. We screened the transformants on MacConkey maltose plates. A colony was isolated that appeared to be resistant to maltose and was pink on these plates, but it was still unable to grow on minimal medium with maltose as the carbon source. The plasmid was isolated, and the gene causing this phenotype was characterized. The deduced amino acid sequence of the encoded protein shows homology to that of lipases and esterases. We termed the gene aes, for acetyl esterase. Extracts of cells harboring plasmid-encoded aes under its own promoter exhibit a fivefold higher capacity to hydrolyze p-nitrophenyl acetate than do extracts of cells of plasmid-free strains. Similarly, strains harboring plasmid-encoded aes are able to grow on triacetyl glycerol (triacetin) whereas the plasmid-free strains are not. The expression of plasmid-encoded aes resulted in strong repression of the maltose transport genes in malT+ strains (10-fold reduction), but not in a malT(Con) strain which is independent of the inducer. Also, overproduction of MalT counteracted the Aes-dependent repression, indicating a direct interaction between MalT and Aes. PMID- 9401026 TI - Unliganded maltose-binding protein triggers lactose transport in an Escherichia coli mutant with an alteration in the maltose transport system. AB - Escherichia coli accumulates malto-oligosaccharides by the maltose transport system, which is a member of the ATP-binding-cassette (ABC) superfamily of transport systems. The proteins of this system are LamB in the outer membrane, maltose-binding protein (MBP) in the periplasm, and the proteins of the inner membrane complex (MalFGK2), composed of one MalF, one MalG, and two MalK subunits. Substrate specificity is determined primarily by the periplasmic component, MBP. However, several studies of the maltose transport system as well as other members of the ABC transporter superfamily have suggested that the integral inner membrane components MalF and MalG may play an important role in determining the specificity of the system. We show here that residue L334 in the fifth transmembrane helix of MalF plays an important role in determining the substrate specificity of the system. A leucine-to-tryptophan alteration at this position (L334W) results in the ability to transport lactose in a saturable manner. This mutant requires functional MalK-ATPase activity and the presence of MBP, even though MBP is incapable of binding lactose. The requirement for MBP confirms that unliganded MBP interacts with the inner membrane MalFGK2 complex and that MBP plays a crucial role in triggering the transport process. PMID- 9401027 TI - rpbA controls transcription of the constitutive phycocyanin gene set in Fremyella diplosiphon. AB - Three gene sets encode alpha and beta subunits of the phycobiliprotein phycocyanin (PC) in the filamentous cyanobacterium Fremyella diplosiphon. The cpcB1A1 set (encodes PC1) is constitutively expressed, whereas the cpcB2A2 set (encodes PC2) is expressed only in red light and the cpcB3A3 set (encodes PC3) is expressed only during sulfur-limited growth. Primary pigment mutant strain FdBM1 is characterized by elevated levels of PC. DNA hybridization analysis showed that like many pigment mutants in our strain collection, strain FdBM1 harbors an extra genomic copy of endogenous transposon Tn5469. By direct cloning from FdBM1 genomic DNA, the extra copy of Tn5469 was localized to an open reading frame, which we have designated the rpbA gene. Complementation experiments correlated rpbA activity to the phenotype of strain FdBM1. The predicted RpbA protein contains two regions resembling the characterized helix-turn-helix motif which is involved in DNA recognition by many bacterial and phage transcription regulator proteins. RNA hybridization analysis showed that relative to the parental strain Fd33, the level of transcripts from cpcB1A1, but not cpcB2A2 or cpcB3A3, was significantly elevated in strain FdBM1. Introduction of the intact rpbA gene into strain FdBM1 restored the cpcB1A1 transcript level to that of strain Fd33. These results suggest that the rpbA gene product functions in controlling constitutive transcription from the cpcB1A1 gene set, possibly as a DNA-binding transcriptional repressor element. PMID- 9401028 TI - Cloning, functional analysis, and transcriptional regulation of the Bacillus subtilis araE gene involved in L-arabinose utilization. AB - The Bacillus subtilis araR locus (mapped at about 294 degrees on the genetic map) comprises two open reading frames with divergently arranged promoters, the regulatory gene, araR, encoding a repressor, and a partially cloned gene, termed araE by analogy to the Escherichia coli L-arabinose permease gene. Here, we report the cloning and sequencing of the entire araE gene encoding a 50.4-kDa polypeptide. The araE gene is monocistronic (as determined by Northern blot analysis), and its putative product is very similar to a number of prokaryotic proton-linked monosaccharide transporters (the group I family of membrane transport proteins). Insertional inactivation of the araE gene leads to a conditional Ara- phenotype dependent on the concentration of L-arabinose in the medium. Therefore, we assume that araE encodes a permease involved in L-arabinose transport into the cell. The araE promoter region contains -10 and -35 regions (as determined by primer extension analysis) very similar to those recognized by RNA polymerase containing the major vegetative-cell sigma factor sigmaA, and the 35 region of the transcription start point for araE is located 2 bp from the -35 region of the araR gene. Transcriptional studies demonstrated that the expression from the araE promoter is induced by L-arabinose, repressed by glucose, and negatively regulated by AraR. These observations are consistent with a model according to which in the absence of L-arabinose, AraR binds to a site(s) within the araE/araR promoter, preventing transcription from the araE promoter and simultaneously limiting the frequency of initiation from its own promoter; the addition of L-arabinose will allow transcription from the araE promoter and increase the frequency of initiation from the araR promoter. PMID- 9401029 TI - Identification of cysteine and arginine residues essential for the phosphotransacetylase from Methanosarcina thermophila. AB - Phosphotransacetylase catalyzes the following reaction: CoASH + CH3CO2PO3(2-) <==> CH3COSCoA + HPO4(2-) (where CoA is coenzyme A). Based on biochemical characterization of the enzyme from the obligate anaerobe Clostridium kluyveri, a ternary mechanism was proposed in which an unspecified cysteine abstracts a proton from CoASH forming a nucleophilic thiolate anion which attacks acetyl phosphate (J. Henkin and R. H. Abeles, Biochemistry 15:3472-3479, 1976). Heterologous production in Escherichia coli of the phosphotransacetylase from Methanosarcina thermophila, an obligately anaerobic methanoarchaeon, allowed site specific replacements to identify essential residues. All four cysteines present in the sequence were individually replaced with alanine, and the kinetic constants of the altered enzymes were determined. The results indicated that only C159 is essential for activity; however, replacement with serine resulted in a fully active enzyme. Activity of the unaltered phosphotransacetylase was sensitive to N-ethylmaleimide. Inhibition kinetics of altered enzymes indicated that this sensitivity resulted from modification of C312, which is at the active site but itself is nonessential for catalysis. Five arginines were individually replaced with glutamine. Kinetic analysis of the altered enzymes identified R310 as essential for activity. Of the four nonessential for activity, R87 and R133 appear to be involved in binding CoA. PMID- 9401030 TI - Characterization of nicotinamide mononucleotide adenylyltransferase from thermophilic archaea. AB - The enzyme nicotinamide mononucleotide (NMN) adenylyltransferase (EC 2.7.7.1) catalyzes the synthesis of NAD+ and nicotinic acid adenine dinucleotide. It has been purified to homogeneity from cellular extracts of the thermophilic archaeon Sulfolobus solfataricus. Through a database search, a highly significant match was found between its N-terminal sequence and a hypothetical protein coded by the thermophilic archaeon Methanococcus jannaschii MJ0541 open reading frame (GenBank accession no. U67503). The MJ0541 gene was isolated, cloned into a T7-based vector, and expressed in Escherichia coli cells, yielding a high level of thermophilic NMN adenylyltransferase activity. The expressed protein was purified to homogeneity by a single-step chromatographic procedure. Both the subunit molecular mass and the N-terminal sequence of the pure recombinant protein were as expected from the deduced amino acid sequence of the MJ0541 open reading frame encoded protein. Molecular and kinetic properties of the enzymes from both archaea are reported and compared with those already known for the mesophilic eukaryotic NMN adenylyltransferase. PMID- 9401031 TI - Transduction of envelope stress in Escherichia coli by the Cpx two-component system. AB - Disruption of normal protein trafficking in the Escherichia coli cell envelope (inner membrane, periplasm, outer membrane) can activate two parallel, but distinct, signal transduction pathways. This activation stimulates the expression of a number of genes whose products function to fold or degrade the mislocalized proteins. One of these signal transduction pathways is a two-component regulatory system comprised of the histidine kinase CpxA and the response regulator, CpxR. In this study we characterized gain-of-function Cpx* mutants in order to learn more about Cpx signal transduction. Sequencing demonstrated that the cpx* mutations cluster in either the periplasmic, the transmembrane, or the H-box domain of CpxA. Intriguingly, most of the periplasmic cpx* gain-of-function mutations cluster in the central region of this domain, and one encodes a deletion of 32 amino acids. Strains harboring these mutations are rendered insensitive to a normally activating signal. In vivo and in vitro characterization of maltose-binding-protein fusions between the wild-type CpxA and a representative cpx* mutant, CpxA101, showed that the mutant CpxA is altered in phosphotransfer reactions with CpxR. Specifically, while both CpxA and CpxA101 function as autokinases and CpxR kinases, CpxA101 is devoid of a CpxR-P phosphatase activity normally present in the wild-type protein. Taken together, the data support a model for Cpx-mediated signal transduction in which the kinase/phosphatase ratio is elevated by stress. Further, the sequence and phenotypes of periplasmic cpx* mutations suggest that interactions with a periplasmic signaling molecule may normally dictate a decreased kinase/phosphatase ratio under nonstress conditions. PMID- 9401032 TI - Cloning of the RHO1 gene from Candida albicans and its regulation of beta-1,3 glucan synthesis. AB - The Saccharomyces cerevisiae RHO1 gene encodes a low-molecular-weight GTPase. One of its recently identified functions is the regulation of beta-1,3-glucan synthase, which synthesizes the main component of the fungal cell wall (J. Drgonova et al., Science 272:277-279, 1996; T. Mazur and W. Baginsky, J. Biol. Chem. 271:14604-14609, 1996; and H. Qadota et al., Science 272:279-281, 1996). From the opportunistic pathogenic fungus Candida albicans, we cloned the RHO1 gene by the PCR and cross-hybridization methods. Sequence analysis revealed that the Candida RHO1 gene has a 597-nucleotide region which encodes a putative 22.0 kDa peptide. The deduced amino acid sequence predicts that Candida albicans Rho1p is 82.9% identical to Saccharomyces Rho1p and contains all the domains conserved among Rho-type GTPases from other organisms. The Candida albicans RHO1 gene could rescue a S. cerevisiae strain containing a rho1 deletion. Furthermore, recombinant Candida albicans Rho1p could reactivate the beta-1,3-glucan synthesis activities of both C. albicans and S. cerevisiae membranes in which endogenous Rho1p had been depleted by Tergitol NP-40-NaCl treatment. Candida albicans Rho1p was copurified with the beta-1,3-glucan synthase putative catalytic subunit, Candida albicans Gsc1p, by product entrapment. Candida albicans Rho1p was shown to interact directly with Candida albicans Gsc1p in a ligand overlay assay and a cross-linking study. These results indicate that Candida albicans Rho1p acts in the same manner as Saccharomyces cerevisiae Rho1p to regulate beta-1,3-glucan synthesis. PMID- 9401033 TI - Mutational analysis of the three cysteines and active-site aspartic acid 103 of ketosteroid isomerase from Pseudomonas putida biotype B. AB - In order to clarify the roles of three cysteines in ketosteroid isomerase (KSI) from Pseudomonas putida biotype B, each of the cysteine residues has been changed to a serine residue (C69S, C81S, and C97S) by site-directed mutagenesis. All cysteine mutations caused only a slight decrease in the k(cat) value, with no significant change of Km for the substrate. Even modification of the sulfhydryl group with 5,5'-dithiobis(2-nitrobenzoic acid) has almost no effect on enzyme activity. These results demonstrate that none of the cysteines in the KSI from P. putida is critical for catalytic activity, contrary to the previous identification of a cysteine in an active-site-directed photoinactivation study of KSI. Based on the three-dimensional structures of KSIs with and without dienolate intermediate analog equilenin, as determined by X-ray crystallography at high resolution, Asp-103 was found to be located within the range of the hydrogen bond to the equilenin. To assess the role of Asp-103 in catalysis, Asp 103 has been replaced with either asparagine (D103N) or alanine (D103A) by site directed mutagenesis. For D103A mutant KSI there was a significant decrease in the k(cat) value: the k(cat) of the mutant was 85-fold lower than that of the wild-type enzyme; however, for the D103N mutant, which retained some hydrogen bonding capability, there was a minor decrease in the k(cat) value. These findings support the idea that aspartic acid 103 in the active site is an essential catalytic residue involved in catalysis by hydrogen bonding to the dienolate intermediate. PMID- 9401034 TI - Regulation of expression of the pilA gene in Myxococcus xanthus. AB - Type IV pili are required for social gliding motility in Myxococcus xanthus. In this work, the expression of pilin (the pilA gene product) during vegetative growth and fruiting-body development was examined. A polyclonal antibody against the pilA gene product (prepilin) was prepared, along with a pilA-lacZ fusion, and was used to assay expression of pilA in M. xanthus in different mutant backgrounds. pilA expression required the response regulator pilR but was negatively regulated by the putative sensor kinase pilS. pilA expression did not require pilB, pilC, or pilT. pilA was also autoregulated; a mutation which altered an invariant glutamate five residues from the presumed prepilin processing site eliminated this autoregulation, as did a deletion of the pilA gene. Primer extension and S1 nuclease analysis identified a sigma54 promoter upstream of pilA, consistent with the homology of pilR to the NtrC family of response regulators. Expression of pilA was found to be developmentally regulated; however, the timing of this expression pattern was not entirely dependent on pilS or pilR. Finally, pilA expression was induced by high nutrient concentrations, an effect that was also not dependent on pilS or pilR. PMID- 9401036 TI - Evidence of participation of McrB(S) in McrBC restriction in Escherichia coli K 12. AB - The McrBC restriction system has the ability to restrict DNA containing 5 hydroxymethylcytosine, N4-methylcytosine, and 5-methylcytosine at specific sequences. The mcrB gene produces two gene products. The complete mcrB open reading frame produces a 51-kDa protein (McrB(L)) and a 33-kDa protein (McrB(S)). The smaller McrB polypeptide is produced from an in-frame, internal translational start site in the mcrB gene. The McrB(S) sequence is identical to that of McrB(L) except that it lacks 161 amino acids present at the N-terminal end of the latter protein. It has been suggested that McrB(L) is the DNA binding restriction subunit. The function of McrB(S) is unknown, although there has been speculation that it plays some role in the modulation of McrBC restriction. Studies of the function of McrB(S) have been challenging since it is produced in frame with McrB(L). In this study, we tested the effects of underproduction (via antisense RNA) and overproduction (via gene dosage) of mcrBC gene products on restriction levels of the mcrBC+ strain JM107. Among the parameters monitored was the induction of SOS responses, which indicate of DNA damage. Evidence from this study suggests that McrB(S) is necessary for stabilization of the McrBC restriction complex in vivo. PMID- 9401035 TI - Myxococcus xanthus sasS encodes a sensor histidine kinase required for early developmental gene expression. AB - Initiation of Myxococcus xanthus multicellular development requires integration of information concerning the cells' nutrient status and density. A gain-of function mutation, sasB7, that bypasses both the starvation and high cell density requirements for developmental expression of the 4521 reporter gene, maps to the sasS gene. The wild-type sasS gene was cloned and sequenced. This gene is predicted to encode a sensor histidine protein kinase that appears to be a key element in the transduction of starvation and cell density inputs. The sasS null mutants express 4521 at a basal level, form defective fruiting bodies, and exhibit reduced sporulation efficiencies. These data indicate that the wild-type sasS gene product functions as a positive regulator of 4521 expression and participates in M. xanthus development. The N terminus of SasS is predicted to contain two transmembrane domains that would locate the protein to the cytoplasmic membrane. The sasB7 mutation, an E139K missense mutation, maps to the predicted N-terminal periplasmic region. The C terminus of SasS contains all of the conserved residues typical of the sensor histidine protein kinases. SasS is predicted to be the sensor protein in a two-component system that integrates information required for M. xanthus developmental gene expression. PMID- 9401037 TI - Cloning and expression of a gene from Streptomyces scabies encoding a putative pathogenicity factor. AB - We cloned a 9.4-kb DNA fragment from Streptomyces scabies ATCC 41973 that allows the nonpathogen Streptomyces lividans 66 TK24 to necrotize and colonize potato tuber slices and produce scab-like symptoms on potato minitubers. Deletion analysis demonstrated that activity was conferred by a 1.6-kb DNA region. Sequence analysis of a 2.4-kb DNA fragment spanning the DNA region necessary for activity revealed three open reading frames (ORFs). The deduced amino acid sequence of ORF1, designated ORFtnp, showed high levels of identity with the first 233 amino acids of the putative transposases of the IS1164 elements from Rhodococcus rhodochrous (71%) and Mycobacterium bovis (68%), members of the Staphylococcus aureus IS256 family of transposases. No significant homologies to ORF2 and ORF3 were found in the nucleic acid and protein databases. ORFtnp is located 5' of ORF3. ORF2 is incomplete and is located 3' of ORF3. Subcloning of the individual ORFs demonstrated that ORF3, designated nec1, is sufficient for necrotizing activity in S. lividans 66 TK24. S. lividans 66 TK24 expressing nec1 does not produce thaxtomin A but produces an unidentified extracellular water soluble compound that causes necrosis on potato tuber discs. The G+C content of nec1 suggests that it has moved horizontally from another genus. Southern analysis of ORFtnp and nec1 demonstrate that these genes are physically linked in Streptomyces strains, including S. scabies and Streptomyces acidiscabies strains, that are pathogenic on potato and that produce the phytotoxin thaxtomin A. These data suggest that nec1 may have been mobilized into S. scabies through a transposition event mediated by ORFtnp. PMID- 9401038 TI - A glgC gene essential only for the first of two spatially distinct phases of glycogen synthesis in Streptomyces coelicolor A3(2). AB - By using a PCR approach based on conserved regions of ADP-glucose pyrophosphorylases, a glgC gene was cloned from Streptomyces coelicolor A3(2). The deduced glgC gene product showed end-to-end relatedness to other bacterial ADP-glucose pyrophosphorylases. The glgC gene is about 1,000 kb from the leftmost chromosome end and is not closely linked to either of the two glgB genes of S. coelicolor, which encode glycogen branching enzymes active in different locations in differentiated colonies. Disruption of glgC eliminated only the first of two temporal peaks of ADP-glucose pyrophosphorylase activity and glycogen accumulation and prevented cytologically observable glycogen accumulation in the substrate mycelium of colonies (phase I), while glycogen deposition in young spore chains (phase II) remained readily detectable. The cloned glgC gene therefore encodes an ADP-glucose pyrophosphorylase essential only for phase I (and it is therefore named glgCI). A second, phase II-specific, glgC gene should also exist in S. coelicolor, though it was not detected by hybridization analysis. PMID- 9401039 TI - Characteristics of Fps1-dependent and -independent glycerol transport in Saccharomyces cerevisiae. AB - Eadie-Hofstee plots of glycerol uptake in wild-type Saccharomyces cerevisiae W303 1A grown on glucose showed the presence of both saturable transport and simple diffusion, whereas an fps1delta mutant displayed only simple diffusion. Transformation of the fps1delta mutant with the glpF gene, which encodes glycerol transport in Escherichia coli, restored biphasic transport kinetics. Yeast extract-peptone-dextrose-grown wild-type cells had a higher passive diffusion constant than the fps1delta mutant, and ethanol enhanced the rate of proton diffusion to a greater extent in the wild type than in the fps1delta mutant. In addition, the lipid fraction of the fps1delta mutant contained a lower percentage of phospholipids and a higher percentage of glycolipids than that of the wild type. Fps1p, therefore, may be involved in the regulation of lipid metabolism in S. cerevisiae, affecting membrane permeability in addition to fulfilling its specific role in glycerol transport. Simultaneous uptake of glycerol and protons occurred in both glycerol- and ethanol-grown wild-type and fps1delta cells and resulted in the accumulation of glycerol at an inside-to-outside ratio of 12:1 to 15:1. Carbonyl cyanide m-chlorophenylhydrazone prevented glycerol accumulation in both strains and abolished transport in the fps1delta mutant grown on ethanol. Likewise, 2,4-dinitrophenol inhibited transport in glycerol-grown wild-type cells. These results indicate the presence of an Fps1p-dependent facilitated diffusion system in glucose-grown cells and an Fps1p-independent proton symport system in derepressed cells. PMID- 9401040 TI - Purification, properties, and sequence of glycerol trinitrate reductase from Agrobacterium radiobacter. AB - Glycerol trinitrate (GTN) reductase, which enables Agrobacterium radiobacter to utilize GTN and related explosives as sources of nitrogen for growth, was purified and characterized, and its gene was cloned and sequenced. The enzyme was a 39-kDa monomeric protein which catalyzed the NADH-dependent reductive scission of GTN (Km = 23 microM) to glycerol dinitrates (mainly the 1,3-isomer) with a pH optimum of 6.5, a temperature optimum of 35 degrees C, and no dependence on metal ions for activity. It was also active on pentaerythritol tetranitrate (PETN), on isosorbide dinitrate, and, very weakly, on ethyleneglycol dinitrate, but it was inactive on isopropyl nitrate, hexahydro-1,3,5-trinitro-1,3,5-triazine, 2,4,6 trinitrotoluene, ammonium ions, nitrate, or nitrite. The amino acid sequence deduced from the DNA sequence was homologous (42 to 51% identity and 61 to 69% similarity) to those of PETN reductase from Enterobacter cloacae, N ethylmaleimide reductase from Escherichia coli, morphinone reductase from Pseudomonas putida, and old yellow enzyme from Saccharomyces cerevisiae, placing the GTN reductase in the alpha/beta barrel flavoprotein group of proteins. GTN reductase and PETN reductase were very similar in many respects except in their distinct preferences for NADH and NADPH cofactors, respectively. PMID- 9401041 TI - Characterization of a DNA polymerase from the uncultivated psychrophilic archaeon Cenarchaeum symbiosum. AB - Cenarchaeum symbiosum, an archaeon which lives in specific association with a marine sponge, belongs to a recently recognized nonthermophilic crenarchaeotal group that inhabits diverse cold and temperate environments. Nonthermophilic crenarchaeotes have not yet been obtained in laboratory culture, and so their phenotypic characteristics have been inferred solely from their ecological distribution. Here we report on the first protein to be characterized from one of these organisms. The DNA polymerase gene of C. symbiosum was identified in the vicinity of the rRNA operon on a large genomic contig. Its deduced amino acid sequence is highly similar to those of the archaeal family B (alpha-type) DNA polymerases. It shared highest overall sequence similarity with the crenarchaeal DNA polymerases from the extreme thermophiles Sulfolobus acidocaldarius and Pyrodictium occultum (54% and 53%, respectively). The conserved motifs of B (alpha-)-type DNA polymerases and 3'-5' exonuclease were identified in the 845 amino-acid sequence. The 96-kDa protein was expressed in Escherichia coli and purified with affinity tags. It exhibited its highest specific activity with gapped-duplex (activated) DNA as the substrate. Single-strand- and double-strand dependent 3'-5' exonuclease activity was detected, as was a marginal 5'-3' exonuclease activity. The enzyme was rapidly inactivated at temperatures higher than 40 degrees C, with a half-life of 10 min at 46 degrees C. It was found to be less thermostable than polymerase I of E. coli and is substantially more heat labile than its most closely related homologs from thermophilic and hyperthermophilic crenarchaeotes. Although phylogenetic studies suggest a thermophilic ancestry for C. symbiosum and its relatives, our biochemical analysis of the DNA polymerase is consistent with the postulated nonthermophilic phenotype of these crenarchaeotes, to date inferred solely from their ecological distribution. PMID- 9401042 TI - Synthesis of nitric oxide from the two equivalent guanidino nitrogens of L arginine by Lactobacillus fermentum. AB - Ten strains of Lactobacillus fermentum that differed in origin converted metmyoglobin to nitrosylmyoglobin [a pentacoordinate nitric oxide (NO) complex of Fe(II) myoglobin] in MRS broth at pH 4.3. Of the 10 strains, L. fermentum IFO 3956 possessed the strongest capacity to convert metmyoglobin to nitrosylmyoglobin. This strain synthesizes NO enzymatically from the two equivalent guanidino nitrogens of L-arginine. To our knowledge, this demonstrates for the first time the production of NO synthesized from the guanidino nitrogens of L-arginine by lactic acid bacteria. IFO 3956 may possess a bacterial NO synthase. PMID- 9401043 TI - bldA-dependent expression of the Streptomyces exfoliatus M11 lipase gene (lipA) is mediated by the product of a contiguous gene, lipR, encoding a putative transcriptional activator. AB - Extracellular lipase synthesis by Streptomyces lividans 66 carrying the cloned lipase gene (lipA) from Streptomyces exfoliatus M11 was found to be growth phase dependent, since lipase was secreted into the medium mainly during the stationary phase; S1 nuclease protection experiments revealed abundant lipA transcripts in RNA preparations obtained during the stationary phase but not in those obtained during exponential growth. Transcription from the lipA promoter was dependent on the presence of lipR, a contiguous downstream gene with a very high guanine-plus cytosine content (80.2%). The deduced lipR product consists of a protein of 934 amino acids that shows similarity to known transcriptional activators and has a strong helix-turn-helix motif at its C terminus; this motif is part of a domain homologous to DNA-binding domains of bacterial regulators of the UhpA/LuxR superfamily. The lipR sequence revealed the presence of a leucine residue, encoded by the rare TTA codon, which caused bldA dependence of lipA transcription in Streptomyces coelicolor A3(2); replacement of the TTA codon by the alternate CTC leucine codon alleviated bidA dependence but not the apparent growth phase dependent regulation of lipA transcription. When lipR expression was induced in a controlled fashion during the exponential growth phase, by placing it under the inducible tipA promoter, lipase synthesis was shifted to the exponential growth phase, indicating that the timing of lipR expression, and not its bldA dependence, is the main cause for stationary-phase transcription of lipA. PMID- 9401044 TI - A Mycobacterium smegmatis mutant with a defective inositol monophosphate phosphatase gene homolog has altered cell envelope permeability. AB - A bacteriophage infection mutant (strain LIMP7) of Mycobacterium smegmatis was isolated following transposon mutagenesis. The mutant showed an unusual phenotype, in that all phages tested produced larger plaques on this strain compared to the parent strain. Other phenotypic characteristics of the mutant were slower growth, increased clumping in liquid culture, increased resistance to chloramphenicol and erythromycin, and increased sensitivity to isoniazid and several beta-lactam antibiotics. Permeability studies showed decreases in the accumulation of lipophilic molecules (norfloxacin and chenodeoxycholate) and a small increase with hydrophilic molecules (cephaloridine); taken together, these characteristics indicate an altered cell envelope. The DNA adjacent to the transposon in LIMP7 was cloned and was shown to be highly similar to genes encoding bacterial and mammalian inositol monophosphate phosphatases. Inositol is important in mycobacteria as a component of the major thiol mycothiol and also in the cell wall, with phosphatidylinositol anchoring lipoarabinomannan (LAM) in the cell envelope. In LIMP7, levels of phosphatidylinositol dimannoside, the precursor of LAM, were less than half of those in the wild-type strain, confirming that the mutation had affected the synthesis of inositol-containing molecules. The impA gene is located within the histidine biosynthesis operon in both M. smegmatis and Mycobacterium tuberculosis, lying between the hisA and hisF genes. PMID- 9401045 TI - Regulation of ornithine utilization in Pseudomonas aeruginosa (PAO1) is mediated by a transcriptional regulator, OruR. AB - We have used transpositional mutagenesis of a proline auxotroph (PAO951) to isolate an ornithine utilization (oru) mutant of Pseudomonas aeruginosa (PAO951 4) that was unable to use ornithine efficiently as the sole carbon and nitrogen source. DNA sequence analysis of the inactivated locus confirmed that the transposon had inserted into a locus whose product demonstrated significant primary sequence homology to members of the AraC family of transcriptional activators. DNA mobility shift assays affirmed this potential regulatory function and indicated that the inactivated gene encodes a transcriptional regulator, which has been designated OruR. In trying to define the ornithine utilization phenotype further, a similar inactivation was engineered in the wild-type strain, PAO1. The resulting isolate (PAO1R4) was totally unable to use ornithine as the sole carbon source. Despite the intensified phenotype, this isolate failed to demonstrate significant changes in any of the catabolic or anabolic enzymes that are known to be subject to regulation by the presence of either ornithine or arginine. It did, however, show modified levels of an enzyme, ornithine acetyltransferase (OAcT), that was previously thought to have merely an anaplerotic activity. Definition of this oruR locus and its effects upon OAcT activity provide evidence that control of ornithine levels in P. aeruginosa may have a significant impact upon how the cell is able to monitor and regulate the use of arginine and glutamate as sources of either carbon or nitrogen. PMID- 9401047 TI - Cloning and characterization of the major outer membrane protein gene (ompH) of Pasteurella multocida X-73. AB - The major outer membrane protein (OmpH) of Pasteurella multocida X-73 was purified by selective extraction with detergents, followed by size exclusion chromatography. The planar lipid bilayer assay showed that OmpH has pore-forming function. The average single channel conductance in 1.0 M KCl was 0.62 nS. The gene (ompH) encoding OmpH has been isolated and sequenced by construction of a genomic library and PCR techniques. The coding region of this gene is 1,059 bp long. The predicted primary protein is composed of 353 amino acids, with a 20 amino-acid signal peptide. The mature protein is composed of 333 amino acids with a molecular mass of 36.665 kDa. The ompH gene encoding mature protein has been expressed in Escherichia coli by using a regulatable expression system. The ompH gene was distributed among 15 P. multocida serotypes and strain CU. Protection studies showed that OmpH was able to induce homologous protection in chickens. These findings demonstrate that OmpH is a protective outer membrane porin of strain X-73 and is conserved among P. multocida somatic serotypes. PMID- 9401046 TI - Cloning and genetic and sequence analyses of the bacteriocin 21 determinant encoded on the Enterococcus faecalis pheromone-responsive conjugative plasmid pPD1. AB - The pheromone-responsive conjugative plasmid pPD1 (59 kb) of Enterococcus faecalis encodes the bacteriocin 21 (bac21) determinant. Cloning, transposon insertion mutagenesis and sequence analysis of the bac21 determinant showed that an 8.5-kb fragment lying between kb 27.1 and 35.6 of the pPD1 map is required for complete expression of the bacteriocin. The 8.5-kb fragment contained nine open reading frames (ORFs), bacA to bac1, which were oriented in the same (upstream-to downstream) direction. Transposon insertions into the bacA to bacE ORFs, which are located in the proximal half of bac21, resulted in defective bacteriocin expression. Insertions into the bacF to bac1 ORFs, which are located in the distal half of bac21, resulted in reduced bacteriocin expression. Deletion mutant analysis of the cloned 8.5-kb fragment revealed that the deletion of segments between kb 31.6 and 35.6 of the pPD1 map, which contained the distal region of the determinant encoding bacF to bac1, resulted in reduced bacteriocin expression. The smallest fragment (4.5 kb) retaining some degree of bacteriocin expression contained the bacA to bacE sequences located in the proximal half of the determinant. The cloned fragment encoding the 4.5-kb proximal region and a Tn916 insertion mutant into pPD1 bacB trans-complemented intracellularly to give complete expression of the bacteriocin. bacA encoded a 105-residue sequence with a molecular mass of 11.1 kDa. The deduced BacA protein showed 100% homology to the broad-spectrum antibiotic peptide AS-48, which is encoded on the E. faecalis conjugative plasmid pMB2 (58 kb). bacH encoded a 195-residue sequence with a molecular mass of 21.9 kDa. The deduced amino acid sequence showed significant homology to the C-terminal region of HlyB (31.1% identical residues), a protein located in the Escherichia coli alpha-hemolysin operon that is a representative bacterial ATP-binding cassette export protein. PMID- 9401048 TI - msDNA-Ec48, the smallest multicopy single-stranded DNA from Escherichia coli. AB - Previously we have reported a novel bacterial reverse transcriptase (RT) from Escherichia coli ECOR58 strains in which the YXDD box was replaced with LVDD (J. R. Mao, S. Inouye, and M. Inouye, Biochem. Biophys. Res. Commun. 227:489-493, 1996). Here we determined the structure of the multicopy single-stranded DNA (msDNA) produced by the RT. The msDNA was found to consist of a single-stranded DNA of 48 nucleotides in length, the shortest msDNA thus far identified from natural sources. The msDNA, the RT, and the retron are designated msDNA-Ec48, RT Ec48, and retron-Ec48, respectively. On the basis of the structure of the msr gene, the RNA molecule of msDNA-Ec48 is predicted to be composed of 119 ribonucleotides; it is the longest RNA among the known msDNAs. Analysis of the DNA sequences flanking the retron indicates that retron-Ec48 is associated with a prophage related to phages P2 and P4. PMID- 9401049 TI - Comparison of the bacterial HelA protein to the F508 region of the cystic fibrosis transmembrane regulator. AB - The HelA protein of Rhodobacter capsulatus is the ATP-binding-cassette subunit of an exporter complex required for cytochrome c biogenesis. By primary sequence comparisons the F88 residue of HelA is similar to the F508 residue of the cystic fibrosis transmembrane regulator (CFTR) protein. Previous studies have established that CFTR F508delta or F508R proteins are defective but F508C is functional. Our results demonstrate that the HelA F88 mutants functionally mimic the phenotypes of known CFTR F508 mutants. The phenotypes of additional HelA mutants and the in vivo steady-state levels of these proteins are also reported. PMID- 9401050 TI - Dinitrogenase reductase-activating glycohydrolase can be released from chromatophores of Rhodospirillum rubrum by treatment with MgGDP. AB - Dinitrogenase reductase-activating glycohydrolase (DRAG), involved in the regulation of nitrogenase activity in Rhodospirillum rubrum, is associated with chromatophore membranes in cell extracts. We show that DRAG can be specifically released by treatment with MgGDP; other nucleotides studied had no effect. The DRAG activity released corresponds to the release of DRAG protein. PMID- 9401051 TI - Inner membrane efflux components are responsible for beta-lactam specificity of multidrug efflux pumps in Pseudomonas aeruginosa. AB - A major feature of the MexAB-OprM multidrug efflux pump which distinguishes it from the MexCD-OprJ and MexEF-OprN multidrug efflux systems in Pseudomonas aeruginosa is its ability to export a wide variety of beta-lactam antibiotics. Given the periplasmic location of their targets it is feasible that beta-lactams exit the cell via the outer membrane OprM without interaction with MexA and MexB, though the latter appear to be necessary for OprM function. To test this, chimeric MexAB-OprJ and MexCD-OprM efflux pumps were reconstituted in delta mexCD delta oprM and delta mexAB delta oprJ strains, respectively, and the influence of the exchange of outer membrane components on substrate (i.e., beta-lactam) specificity was assessed. Both chimeric pumps were active in antibiotic efflux, as evidenced by their contributions to resistance to a variety of antimicrobial agents, although there was no change in resistance profiles relative to the native pumps, indicating that OprM is not the determining factor for the beta lactam specificity of MexAB-OprM. Thus, one or both of inner membrane-associated proteins MexA and MexB are responsible for drug recognition, including recognition of beta-lactams. PMID- 9401053 TI - Hepatitis C virus-related lymphomas. PMID- 9401052 TI - Bordetella bronchiseptica expresses the fimbrial structural subunit gene fimA. AB - The differential host species specificities of Bordetella pertussis, B. parapertussis, and B. bronchiseptica might be explained by polymorphisms in adherence factor genes. We have found that B. parapertussis and B. bronchiseptica, unlike B. pertussis, contain a full-length gene for the fimbrial subunit FimA. B. bronchiseptica expresses fimA in a BvgAS-dependent fashion. PMID- 9401054 TI - Induction of nitric oxide synthase is involved in the mechanism of Fas-mediated apoptosis in haemopoietic cells. AB - Induction of nitric oxide synthase (iNOS) and production of the toxic metabolite nitric oxide (NO) is one of the interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF-alpha) regulated effector mechanisms that can lead to apoptosis of haemopoietic progenitor cells. Fas-receptor (Fas-R) expression can be stimulated by IFN-gamma and TNF-alpha. Transactivation of iNOS, and possibly Fas R promoters, by interferon regulatory factor-1 expressed in response to IFN-gamma may be a part of the iNOS transduction pathway. We investigated whether the effects of Fas-R triggering in haemopoietic cells were mediated by NO. On Western blotting, we observed that Fas-receptor agonist, monoclonal antibody CH11. enhanced expression of iNOS. As shown by the reverse transcription polymerase chain reaction. CH11 also induced iNOS mRNA expression in purified CD34+ cells. To determine whether NO was involved in Fas-mediated apoptosis we inhibited iNOS catalysed production of NO using anti-sense (AS) oligodeoxynucleotides (ODN) directed against iNOS mRNA. After culture of haemopoietic cells in the presence of AS-ODN, iNOS expression decreased and was no longer enhanced by Fas. This effect was associated with the prevention of Fas-mediated apoptosis, as determined by a DNA fragmentation and terminal deoxynucleotidyl transferase staining. In colony assays, specific AS-oligonucleotides prevented FAS-mediated inhibition of colony formation by total bone marrow and CD34+ progenitor cells. Our data suggest that the inhibitory effects of Fas, including induction of apoptosis, are mediated by effector mechanisms that may be similar to those described for IFN-gamma and TNF-alpha. PMID- 9401055 TI - CD34+ peripheral blood progenitor cell and monocyte derived dendritic cells: a comparative analysis. AB - Dendritic cells (DC) have been generated in vitro from either CD34+ haemopoietic progenitor cells (HPC) or peripheral blood monocytes (Mo) in the presence of specific cytokine combinations, including granulocyte-macrophage colony stimulating factor (GM-CSF). Since differences between DC from either source may be important for the clinical use of these antigen-presenting cells (APC), a comparative analysis was performed. HPC were expanded in the presence of interleukin (IL)-3, IL-6 and stem cell factor (SCF) (days 1-7) and subsequently induced by IL-4+ GM-CSF (days 8-26) to differentiate to Langerhans-type cells (pLC). The latter cytokines were similarly used to generate Mo-derived LC (mLC). Maturation of both cell types, pLC and mLC, to interdigitating DC-type cells (iDC) was induced by tumour necrosis factor-alpha (TNF-alpha) or lipopolysaccharide (LPS). Analysis of mLC/pLC and miDC/piDC with respect to morphology, phenotype, antigen uptake and presentation revealed a high similarity of DC from either source. The majority of mLC, however, exhibited a more mature differentiation stage, compared to pLC, evidenced from lower numbers of multilaminar MHC class II compartments and less efficient APC function for extracellular protein antigens. Although macropinocytosis was performed by LC, neither LC nor iDC from either source were able to take up > or = 0.5 microm latex beads. However, phagocytosis of 0.5 microm and 1 microm beads was performed by Mo that could subsequently be induced to become iDC, thus providing the unique opportunity to present phagocytosed material in DC-type fashion. Mo may be the preferential source for clinical use of iDC-type cells since preparation and culture are easier to perform and are less costly while APC function is similar to HPC-derived iDC. PMID- 9401056 TI - Constitutive and inducible expression of megakaryocyte-specific genes in Friend erythroleukaemia cells. AB - Friend murine erythroleukaemia cells (MELC) were analysed by semiquantitative RT PCR for the constitutive and inducible expression of megakaryocyte-specific genes. Uninduced MELC expressed detectable levels of mRNAs for acethylcholinesterase (AChE), platelet factor-4 (PF4), glycoprotein IIb (GPIIb) and von Willebrand factor (VWF), whereas the erythroid alpha- and beta-globin genes were not transcribed appreciably. However, MELC exposed to 5 mM hexamethylene bisacetamide (HMBA) or 1.5% dimethyl sulphoxide (DMSO) seemed to be channelled towards a mixed erythroid/megakaryocytic phenotype characterized by unaltered levels of VWF mRNA, increased levels of AChE, GPIIb and PF4 mRNA. and simultaneous induction of the globin genes. Megakaryocyte-related genes were expressed. in the absence of globin gene transcription, by MELC treated with either phorbol-12-myristate acetate (PMA; 100 ng/ml) or colcemid (40 nM), an antimicrotubule agent capable of promoting polyploidization in this model. Moreover, PMA and colcemid induced also de novo expression of the thrombopoietin receptor c-mpl. PMA and colcemid did not affect high basal c-myb mRNA levels which, in turn, were down-regulated upon HMBA or DMSO induction. Additionally, both uninduced and induced MELC exhibited significant levels of Epo-R and IL-3R mRNAs, whereas no expression of granulocyte/macrophage-related genes was detected. Megakaryocyte gene expression of MELC was also compared to that of other haemopoietic cell populations from normal mice and mice infected with the anaemic strain of the Friend virus. According to our results, MELC should be seen as an unique erythro-megakaryocytic model of differentiation, potentially useful for studying molecular events governing lineage commitment as well as some steps of megakaryocytopoiesis. PMID- 9401058 TI - T cells selectively infiltrate bone marrow areas with residual haemopoiesis of patients with acquired aplastic anaemia. AB - Aplastic anaemia (AA) is characterized by pancytopenia and bone marrow (BM) hypocellularity. In some patients AA may be mediated by T cells. To localize inflammatory cell infiltrates, we carried out a quantitative immunohistochemical analysis of BM biopsies of AA patients. In five out of eight biopsies, significantly higher numbers T cells were found in the areas with residual haemopoiesis (RH). The significantly increased numbers of CD3+ T cells in areas with RH supports the hypothesis of a site-directed infiltration and/or a local proliferation of T cells in the BM of patients with AA. PMID- 9401057 TI - Megakaryocytes derived from CD34-positive cord blood cells produce interleukin-8. AB - In a serum-free liquid culture, thrombopoietin (TPO) selectively stimulated the growth of megakaryocytic cells from CD34-positive cord blood cells. Using these cultured cells, we investigated cytokine production by human megakaryocytes. Day 10 megakaryocytes (2 x 10(5)) secreted > 1000 pg/ml of interleukin (IL)-8, in contrast to small amounts of IL-1beta and IL-6. A time-course study showed that the IL-8 production of megakaryocytes occurred at the late phase of the culture period. The megakaryocyte-conditioned medium had the chemotactic potential of polymorphonuclear leucocytes, which was abrogated by the addition of anti-IL-8 antibody, suggesting the secretion of biologically active IL-8. The combination of TPO and IL-1alpha was required for a significant augmentation of the IL-8 secretion. Direct evidence for IL-8 synthesis in megakaryocytes was provided by reverse transcription-polymerase chain reaction on purified CD41b+ cells and by the detection of intracellular IL-8 in CD41b+ cells. These results suggest that TPO stimulates not only the proliferation and differentiation of the progenitors capable of megakaryocytic lineage expression but also IL-8 release by the megakaryocytic cells with the aid of IL-1. PMID- 9401059 TI - Severe aplastic anaemia in association with a unique constitutional translocation 46,XY,t(6;10)(q13;q22)c. AB - Severe aplastic anaemia (SAA) is an uncommon disorder which may be associated with several congenital syndromes. However, it has rarely been described in association with a constitutional karyotypic abnormality. The breakpoint of the balanced t(6:10)(q13:q22) translocation described here does not disrupt any currently recognized gene of haemopoietic or stromal importance. This report also highlights the problems inherent in the use of bone marrow transplantation (BMT) for treating multiply transfused aplastic anaemia patients. PMID- 9401060 TI - Total absence of protein 4.2 and partial deficiency of band 3 in hereditary spherocytosis. AB - Unlike previously reported cases with total protein 4.2 deficiency due to mutations in the EPB42 gene, we describe a total deficiency in protein 4.2 with normal EPB42 alleles. Hereditary spherocytosis (HS) was observed in a Japanese woman (unsplenectomized) and her daughter (splenectomized). The mother showed a partial deficiency in band 3 and a proportional reduction in protein 4.2. She was heterozygous for a novel allele of the EPB3 gene, allele Okinawa, which contains the two mutations that define the Memphis II polymorphism (K56E, AAG-->GAG, and P854L, CCG-->CTG) and, additionally, the mutation: G714R, GGG-->AGG, located in a highly conserved position of transmembrane segment 9. The latter change was responsible for HS. In trans to allele Okinawa, the daughter displayed allele Fukuoka: G130R, GGA-->AGA, an allele known to alter the binding of protein 4.2 to band 3. The daughter presented with a more pronounced decrease of band 3, and lacked protein 4.2, resulting in aggravated haemolytic features. Although the father was not available for study, heterozygosity for allele Fukuoka has been documented in another individual who showed no clinical or haematological signs, and a normal content of band 3. We suggest that band 3 Okinawa binds virtually all the protein 4.2 in red cell precursors, band 3 Fukuoka being unable to do so, and that the impossibility of band 3 Okinawa incorporation into the membrane leads to degradation of the band 3 Okinawa protein 4.2 complex. In contrast, band 3 Fukuoka, free of bound protein 4.2, could then incorporate normally into the bilayer. Thus, protein 4.2 would not appear in the daughter's red cell membrane. PMID- 9401061 TI - Distribution of the cytogenetic abnormality +i(3)(q10) in persistent polyclonal B cell lymphocytosis: a FICTION study in three cases. AB - Persistent polyclonal B-cell lymphocytosis (PPBL) is a rare entity characterized by a moderate but sustained lymphocytosis where some binucleated or bilobulated circulating forms constitute, even if they are not entirely specific, the cytological hallmark of the disease. An additional chromosome long arm i(3)(q10) has recently been reported as a recurrent cytogenetic aberration, contrasting with a usual polyclonal immunoglobulin expression. To determine more precisely the distribution of the chromosomal abnormality within the peripheral lymphocyte population and study the relationship between the +i(3)(q10) and the bilobulated character, we investigated three new cases of PPBL displaying the cytogenetic abnormality on the karyotype, using a technique of simultaneous fluorescence immunophenotyping and interphase cytogenetics (FICTION). We demonstrated that the +i(3)(q10) was restricted to the B lymphocytes, independently of the kappa or lambda light chain isotype and was present in both bilobulated and non bilobulated cells. Therefore it is likely that the cytogenetic abnormality occurs at an early stage of lymphocyte differentiation in a precursor cell already committed to the B-cell lineage, before any rearrangement of immunoglobulin genes has taken place. PMID- 9401062 TI - Addition of granulocyte colony-stimulating factor to chemotherapy in patients with AIDS-related lymphoma: effects on neutrophil Fc gamma receptor expression and soluble Fc gammaRIII plasma levels. AB - AIDS-related neutropenia and neutrophil dysfunction can (partly) be reversed by granulocyte-colony stimulating factor (G-CSF). We studied the effect of G-CSF on neutrophil increment and levels of soluble Fc gamma receptor type III in 15 patients with AIDS-related lymphoma (ARL) undergoing chemotherapy. In six of these patients we performed a detailed kinetic analysis of the membrane expression of the functionally important Fc gamma-receptors type I, II and III. In all these patients G-CSF induced Fc gammaRI positive neutrophils with a decreased expression of the Fc gammaRIII receptor. These changes were similar to those seen both in healthy volunteers and in non-HIV-infected individuals treated with chemotherapy. Interestingly, the mean neutrophil and sFc gammaRIII increment were significantly lower and more patients had a nadir granulocyte count < 0.5 x 10(9)/l after the first cycle than after the second cycle of chemotherapy. This may be related to a therapy-associated decrease in HIV-1 viral load. The conclusion is that patients treated with chemotherapy for ARL have a qualitatively normal response to G-CSF. PMID- 9401063 TI - Platelet activity of high-dose factor VIIa is independent of tissue factor. AB - High-dose recombinant factor VIIa has been successfully used as therapy for haemophiliacs with inhibitors. The mechanism by which high-dose factor VIIa supports haemostasis is the subject of some controversy. Postulating a mechanism in which activity is dependent on tissue factor at the site of injury explains the localization of activity but not the requirement for high doses. Postulating a mechanism in which factor VIIa acts on available lipid independently of tissue factor explains the requirement for high doses but not the lack of systemic procoagulant activity. We report that factor VIIa bound weakly to activated platelets (Kd approximately 90 nM). This factor VIIa was functionally active and could initiate thrombin generation in the presence of plasma concentrations of prothrombin, factor X, factor V, antithrombin III and tissue factor pathway inhibitor. The activity was not dependent on tissue factor. The concentration of factor VIIa required for detectable thrombin generation agreed well with the lowest concentration of factor VIIa required for efficacy in patients. High-dose factor VIIa may function on the activated platelets that form the initial haemostatic plug in haemophilic patients. These observations are in agreement with clinical trials which have shown that high-dose factor VIIa was haemostatically effective without causing systemic activation of coagulation. PMID- 9401064 TI - The effect of thrombin on the dynamic exchange between intraplatelet and extraplatelet fibrinogen. AB - We examined the distribution of platelet fibrinogen and the exchange between intra- and extra-platelet fibrinogen in unstimulated and thrombin-stimulated platelets. In unstimulated platelets 60% of platelet fibrinogen was found in the soluble platelet fraction and 40% in the insoluble one. In platelets activated with thrombin, changes took place in the distribution of intraplatelet fibrinogen but not in the total fibrinogen content. At > or = 0.5 U/ml of thrombin the fibrin(ogen) content of the insoluble and soluble fractions was approximately 80% and 20%, respectively. When we evaluated how extraplatelet fibrinogen affects the content and distribution of intraplatelet fibrinogen, we found that when unlabelled fibrinogen was added to unstimulated and thrombin-stimulated platelets the content and distribution of intraplatelet fibrinogen remained unaltered. However, when 125I-fibrinogen was added, it was incorporated into unstimulated and thrombin-stimulated platelets. In unstimulated platelets, 70% of the incorporated 125I-fibrinogen was in the soluble fraction and 30% in the insoluble. In thrombin-stimulated platelets the distribution of the incorporated 125I-fibrinogen was 62% and 38% in soluble and insoluble fractions respectively. MoAb to GPIIb-IIIa produced 80% and 60% inhibition of 125I-fibrinogen incorporation by unstimulated and thrombin-stimulated platelets. Our data showed dynamic exchange between intraplatelet and extraplatelet fibrinogen both in unstimulated and thrombin-stimulated platelets mediated mainly by GPIIb-IIIa. PMID- 9401065 TI - Brefeldin A inhibits thrombin receptor regeneration in human endothelial cells. AB - Endothelial cells, once stimulated with thrombin, are resistant to subsequent stimulation. After a recovery period of about 60 min the cells are sensitized again for activation by thrombin. The resensitization is independent of the receptor de novo synthesis. Therefore an intracellular pool of thrombin receptors that is possibly co-localized with the Golgi apparatus has been assumed. Brefeldin A (BFA) has been used extensively to investigate the intracellular sorting of proteins because of its dramatic alteration of the structural and functional organization of the Golgi apparatus. Accordingly we have examined the effects of BFA on the regeneration of the thrombin receptor response in human umbilical vein and artery cells. The von Willebrand factor (VWF) release from Weibel-Palade vesicles and the intracellular calcium mobilization were used as physiological parameters of thrombin receptor activation. The addition of BFA (2 microg/ml) to endothelial cells or the reduction of the incubation temperature from 37 degrees C to 16 degrees C blocked the receptor response regeneration almost completely. PMID- 9401066 TI - Fibrinogen Kaiserslautern (gamma 380 Lys to Asn): a new glycosylated fibrinogen variant with delayed polymerization. AB - An adult woman diagnosed with cerebral thrombosis following a caesarean section was found to have severely prolonged thrombin and reptilase times. Five other family members also had prolonged, but variable, thrombin and reptilase times. Analysis of purified fibrinogen on reducing SDS-PAGE revealed an additional band, in all family members, which migrated immediately below the normal B beta band. Western blotting indicated that this band was a gamma chain and endoglycosidase-F digestion established that it contained an additional oligosaccharide side chain. Partial acid hydrolysis localized the new oligosaccharide to the C-terminus of the gamma chain. Amplification of this region by PCR and subsequent DNA sequencing demonstrated a single base substitution altering the normal 380 Lys (AAG) codon to Asn (AAT), producing a new Asn-Lys-Thr glycosylation site. The propositus and one other family member were homozygous for this mutation but the remaining four family members were heterozygous. The polymerization of purified fibrin monomers from the propositus was grossly abnormal; however, the polymerization curve was almost normalized by the removal of terminal sialic acid residues. This suggests that the polymerization defect was primarily caused by additional negatively charged sialic acid residues present on the new oligosaccharide. Further analysis of the D domain of purified fibrinogen established that calcium binding to the high affinity site remained unaffected by the bulky carbohydrate side chain or negatively charged sialic acid residues. PMID- 9401067 TI - Tumour cell u-PA as a cause of fibrinolytic bleeding in metastatic disease. AB - Tumour cells may express urokinase type plasminogen activator (u-PA). This may influence the invasive properties of the cells but has seldom been implicated in production of a systemic bleeding state. Two patients are described in whom severe bleeding occurred in association with disseminated malignancies. Thrombin generation was little disturbed and platelet numbers were insufficient to account for the bleeding. Florid plasmin generation was evident in the circulation and the fibrinolytic inhibitor tranexamic acid controlled the bleeding well. Free active u-PA was demonstrated in the circulation and u-PA antigen on the malignant cells which invaded the marrow of one of the patients. Tumour cell u-PA may occasionally be responsible for a bleeding state. PMID- 9401068 TI - Severe factor XI deficiency in an Arab family associated with a novel mutation in exon 11. AB - We investigated an 8-year-old Arab girl with severe factor XI deficiency; one sibling and her father also have severe factor XI deficiency. Her parents and her father's parents are first cousins. Restriction analysis and DNA sequencing excluded the type I, II, III and IV mutations. We demonstrated a previously undescribed C-->A mutation at nucleotide 1254 in exon 11 resulting in a threonine to asparagine (T-->N) substitution at amino acid 386. We postulate that this substitution interferes with folding and secretion of the molecule. PMID- 9401069 TI - Clotting factor IX levels in C/EBP alpha knockout mice. AB - Previous studies have indicated that C/EBP alpha is involved in the regulation of factor IX and mutations of a C/EBP recognition element in the factor IX promoter result in haemophilia B. We now report that mice homozygous for the deletion of the c/ebp alpha gene are significantly deficient in factor IX transcription. PMID- 9401070 TI - A comparison of outcome from febrile neutropenic episodes in children compared with adults: results from four EORTC studies. International Antimicrobial Therapy Cooperative Group (IATCG) of the European Organization for Research and Treatment of Cancer (EORTC). AB - The object of this study was to determine whether there were any differences between the 'typical' child with fever and neutropenia and their adult counterpart with regard to infection type and outcome, by analysis of 3080 patients, including 759 children < 18 years of age and 2321 adults. These represented patients randomized in previous trials, between 1986 and 1994, which compared empirical antibiotic regimens for fever in neutropenic patients. There were fewer childhood acute myeloid leukaemia patients than adults but more acute lymphoblastic leukaemia cases and more with solid tumours undergoing intensive myelosuppressive therapy. The children were less likely to be undergoing first induction therapy but the relative incidence of patients receiving relapse schedules or maintenance therapies were not significantly different in the two age groups. Children less frequently had a defined site of infection than adults and where they had a defined site there were more upper respiratory tract but fewer lung infections. There was a similar low incidence of shock at presentation in the two groups but the children's median neutrophil count was lower, and their median duration of granulocytopenia before the trial was shorter. The incidence of bacteraemia was similar, but clinically documented infection was less frequent and fever of unknown origin consequently more common in children. Children developed more streptococcal bacteraemias and fewer staphylococcal bacteraemias than adults (P=0.003) but the relative incidence of various gram-negative species was similar (P=0.57). In general, the children had a better overall success rate and lower mortality than adults. Death from infection was only 1% in children versus 4% in adults (P=0.001), and time to defervescence was shorter in children. In the younger age group, univariate logistic regression models showed high temperature, prolonged neutropenia before the trial and shock as prognostic indicators for the presence of bacteraemia. Solid tumour patients were significantly less likely to have a bacteraemia. Multivariate analysis confirmed the independent prognostic value of these indicators. Using the logistic equation of the selected model, the overall discriminant ability was poor. However, it was possible to identify a small subgroup without shock or high fever and with a short prior duration of neutropenia which carries a particularly low risk of bacteraemia, who could be considered for early discharge, monotherapy and shortened courses of antibodies, in prospective trials. PMID- 9401071 TI - Comparative genomic hybridization is a powerful tool, complementary to cytogenetics, to identify chromosomal abnormalities in childhood acute lymphoblastic leukaemia. AB - Cytogenetics has a strong prognostic value in childhood acute lymphoblastic leukaemia (ALL), but results are often incomplete because of the poor chromosome morphology. To improve this analysis, we tested comparative genomic hybridization (CGH) for the detection of chromosomal imbalances. 72 children were retrospectively analysed using CGH. Only 53% of the patients had been fully banded by standard methods. With CGH, 36 patients retained a normal chromosomal profile and 36 had unbalanced abnormalities. No amplification was detected. Fluorescence in situ hybridization (FISH) with centromeric and unique sequence probes was used in those cases with discrepancies or unsuccessful karyotype to validate CGH results. CGH enabled clear identification of unbalanced chromosomal abnormalities, even in some cases which had a normal karyotype. In view of the strong prognostic value of hyperdiploidy in childhood ALL, CGH appears to be a powerful technique, complementary to conventional cytogenetics. PMID- 9401072 TI - The rapid diagnosis of acute promyelocytic leukaemia using PML (5E10) monoclonal antibody. AB - Acute promyelocytic leukaemia (APL) is characterized cytogenetically by t(15;17)(q22:q21) which results in the production of a PML/RAR alpha fusion protein. Detection of the translocation or the fusion gene product is required for objective diagnosis of APL. This can be accomplished by conventional cytogenetic methods, fluorescence in situ hybridization or RT-PCR. Such techniques are time consuming and not universally available. The intracellular distribution of the PML protein in promyelocytes is characteristically altered in APL and this can be detected by immunocytochemistry. We have assessed two immunocytochemical methods, immunofluorescence and alkaline phosphatase-anti alkaline phosphatase staining (APAAP), with regard to sensitivity, specificity and rapidity of diagnosis. 85 patients with AML including 15 cases of APL were studied. Immunofluorescence PML detection was concordant with RT-PCR for t(15:17) in 14/15 (93.3%) cases with no false positives. The negative APL case in our series was a patient with a 5' PML breakpoint who did not express the reciprocal t(17;15) fusion product. APAAP was concordant in only 6/13 (46%) APL cases with one false positive. In conclusion, immunofluorescent localization of PML using 5E10 monoclonal antibody is a rapid, sensitive and specific diagnostic tool for APL. PMID- 9401073 TI - The significance of trisomy 8 in de novo acute myeloid leukaemia: the accompanying chromosome aberrations determine the prognosis. German AML Cooperative Study Group. AB - Trisomy 8 is the most frequent numerical chromosome aberration in acute myeloid leukaemia (AML). It occurs either as the sole anomaly or together with other clonal chromosome aberrations. We investigated whether accompanying chromosome anomalies influence the clinical outcome in patients with trisomy 8 and de novo AML. Since 1986, in 713 AML cases treated according to the protocols of the German AMLCG trials, chromosome analyses have been successfully performed. The overall incidence of trisomy 8 was 7.6%. Complete clinical follow-up data were available for 51 patients who were divided into three different categories: group 1: trisomy 8 as the sole cytogenetic anomaly (n = 20); group 2: trisomy 8 in addition to favourable chromosome aberrations (t(8;21)(q22;q22), t(15;17)(q22;q21), inv(16)(p13q22)) (n = 10); and group 3: trisomy 8 accompanied by other anomalies, in most cases of complex type (n = 21). Complete remission (CR) rates were 70%, 90% and 67% for groups 1, 2 and 3, respectively. Event-free survival (EFS) at 3 years differed significantly between patients with trisomy 8 only (37.5%), patients with trisomy 8 in combination with favourable aberrations (55.0%) and patients with trisomy 8 and other accompanying anomalies, mostly complex chromosome aberrations (9.0%) (group 1 v group 2: P=0.12; group 1 v group 3: P=0.005; group 2 v group 3: P=0.05). In this study patients with +8 as the sole cytogenetic anomaly had an intermediate prognosis, patients with +8 in addition to favourable chromosome aberrations maintained a good clinical outcome, and patients with +8 in combination with other abnormalities showed the worst prognosis. PMID- 9401074 TI - Aberrant FHIT transcripts in acute myeloid leukaemia. AB - Recently the FHIT gene (fragile histidine triad gene) has been identified at chromosome 3p14.2 and a high frequency of abnormalities in this gene has been demonstrated in various cancers. To determine the role of the FHIT gene in leukaemia, bone marrow or peripheral blood from 62 acute myeloid leukaemia patients and five haemopoietic cell lines (HL60, U937, Raji, KC-1, K562) were analysed by reverse transcription of the FHIT mRNA followed by PCR amplification and sequencing of the products. To detect the deletion of the FHIT gene, 17 cases were evaluated using microsatellite polymorphism analysis. In this study, 17/62 (27%) AML patients expressed aberrant transcripts which lack two or more exons of the FHIT gene, and all the cell lines exhibited the aberrant FHIT transcripts. No cases exhibited a loss of the FHIT alleles. Our data indicated that the FHIT gene may play a role in myeloid carcinogenesis and may be indicated in the late progression of the disease. PMID- 9401075 TI - Activity of TNF-related apoptosis-inducing ligand (TRAIL) in haematological malignancies. AB - T-cell cytotoxicity is primarily mediated by two cell surface proteins, Fas ligand (FasL) and tumour necrosis factor-related apoptosis-inducing ligand (TRAIL), and intracellular perforin and granzyme granules. FasL-deficient and perforin-deficient T lymphocytes maintain cytotoxicity but fail to induce graft versus-host disease (GVHD) when transplanted into mice. suggesting that GVHD and graft-versus-tumour (GVT) effects can be dissociated, and that TRAIL is not involved in the pathogenesis of GVHD. Because TRAIL could mediate a favourable GVT effect it became important to study the spectrum of its activity and to investigate factors that can dissociate its expression from FasL. TRAIL induced apoptosis in 11/41 (27%) tumour specimens of haematological origin compared to 16/41 (39%) induced by FasL. Although eight specimens were sensitive to both FasL and TRAIL, no synergism was observed between these two ligands. TRAIL induced apoptosis in a dose and time dependent manner with an ED50 of 0.5 microg/ml and EDmax of 1 microg/ml. TRAIL activity was not reduced by the over-expression of the multidrug resistant (MDR) protein, and was not enhanced by 9-cis retinoic acid (RA), which can down-regulate bcl-2 protein. Both ligands were simultaneously up-regulated in normal peripheral blood lymphocytes in response to IL-2, IL-15 and anti-CD3 antibody, whereas IL-10 had no effect. Together, our data show that (1) TRAIL can mediate cell death in a variety of human haematological malignancies, (2) resistance to TRAIL is not mediated by MDR protein, (3) the lack of synergy between TRAIL and FasL suggests that either one is sufficient to mediate T-cell cytotoxicity, and (4) within the panel of cytokines tested, the expression of TRAIL and FasL could not be dissociated. PMID- 9401077 TI - Genetic analysis of 8;21 chromosomal translocation without AML1 gene involvement in MDS-AML. AB - We have investigated a case of acute myelocytic leukaemia derived from myelodysplastic syndrome (MDS-AML) with an 8;21 translocation. In this case the AML1/MTG8 (ETO) fusion transcript was not detected by reverse transcriptase polymerase chain reaction (RT-PCR), and the rearrangement of the AML1 gene locus was not detected by Southern blot nor pulse field gel electrophoresis (PFGE) analyses using specific probes for the AML1 gene. Fluorescence in-situ hybridization (FISH) study using cosmid probes for 21q22 revealed that the breakpoint of 21q22 was telomeric to the AML1 gene locus and centromeric from D21S259, 351, 3421 loci. This is the first report concerning the t(8;21)(q22;q22) carrying AMLs (de novo AML, MDS-AML and therapy-related AML) to show that the breakpoint at 21q22 is located outside the AML1 gene locus. It is also noteworthy that the cell-surface antigen expression pattern of the bone marrow (BM) blasts was changed from CD7+ CD2+ CD13+ CD33+ CD19- CD11b+ CD14+ CD36+ to CD7- CD2- CD13+ CD19+ CD11b- CD14- CD33+ CD34+ CD36- CD56+ during leukaemic progression, and the pattern in leukaemic phase was similar to the characteristic phenotype of de novo AML cases with t(8;21), when the AML1/MTG8 fusion transcripts are always detected by RT-PCR. PMID- 9401076 TI - Oxidative DNA damage in CD34+ myelodysplastic cells is associated with intracellular redox changes and elevated plasma tumour necrosis factor-alpha concentration. AB - Ineffective haemopoiesis in the myelodysplastic syndromes (MDS) is mediated, at least in part, by apoptosis, though the mechanisms of apoptotic induction are unclear. Tumour necrosis factor-alpha (TNF-alpha) promotes apoptosis via intracellular oxygen free radical production, oxidation of DNA and proteins, and is increasingly implicated in the pathogenesis of MDS. Using single-cell gel electrophoresis, we have identified oxidized pyrimidine nucleotides in the progenitor-enriched bone marrow CD34+ compartment from MDS patients (P=0.039), which are absent in both CD34- MDS cells (P=0.53) and also CD34+ cells from normal subjects (P=0.55). MDS CD34+ blood cells also showed oxidized pyrimidine nucleotides compared with CD34- cells (P=0.029). Within normal subjects no differences were seen between CD34+ and CD34- bone marrow cell compartments. CD34+ bone marrow cell oxidized pyrimidines were strongly associated with elevated plasma TNF-alpha and low bone marrow mononuclear cell glutathione concentrations (5/6 patients) and the inverse relationship was also found (3/4 patients). This data implies a role for intracellular oxygen free radical production, perhaps mediated by TNF-alpha, in the pathogenesis of ineffective haemopoiesis in MDS and provides a rationale for the bone marrow stimulatory effects of antioxidants such as Amifostine in MDS. PMID- 9401078 TI - Bone marrow fibrosis and disease activity in multiple myeloma monitored by the aminoterminal propeptide of procollagen III in serum. AB - Simple bone marrow fibrosis is seen in 10-30% of multiple myeloma (MM) patients. We investigated the incidence and characteristics of the bone marrow stromal alterations, in order to characterize the collagens involved by immunohistochemistry, and to evaluate the use of serum aminoterminal propeptide of type III procollagen (PIIINP) as a marker of marrow fibrogenesis and disease activity in MM. 34 consecutive patients with newly diagnosed MM were included prospectively, and followed for 12-30 months. Compared with the findings in 15 normal individuals we found increased interstitial deposits of collagen III in 48% of MM patients, whereas deposits of collagen I were not increased. Interstitial fibrosis appeared to be restricted to areas of severe plasma cell infiltration, but it could also have a more dispersed presentation in the severely infiltrated marrow. There was a high co-distribution of collagen III fibrils and reticulin fibres. Serum PIIINP levels were elevated in most patients, and in the follow-up study serum PIIINP showed a good correlation with the response to treatment. Patients with resistant or progressive disease had continually elevated levels of PIIINP. In most patients with responsive disease serum PIIINP normalized, and we observed no relapses in patients who had normal serum PIIINP levels. Other patients who responded to treatment by reduced M component level, but had persistently elevated serum levels of PIIINP, had either early relapses or developed progression of osteolytic lesions in spite of unchanged M-component levels. Therefore an elevated serum PIIINP during treatment might indicate an active malignant clone. Serum PIIINP does not simply follow the M-component, but gives further information of potential therapeutic value. PMID- 9401079 TI - Magnetic resonance imaging patterns in patients with multiple myeloma. AB - Sixty-one consecutive patients with multiple myeloma were studied with magnetic resonance (MR) imaging of the spine. Sagittal T1-weighted and short inversion time (TI) inversion recovery (STIR) images were obtained. The MR patterns of the bone marrow were classified as diffuse (D) (n=26), nodular (N) (n=11), D+N (n=13) or normal (n) (n=11). Abnormal patterns were seen in 50 (82%) of the 61 patients. Correlations were found between the MR imaging patterns and some laboratory findings (WBC, haematocrit, platelet count, serum albumin, and percentage of marrow plasmacytosis). The survival of the patients with abnormal MRI patterns was significantly poorer than that of the patients with normal patterns. However, the survival of patients with a nodular pattern did not differ from those with a normal pattern. The MR imaging pattern of the bone marrow in patients with multiple myeloma is a useful factor in the assessment of prognosis. PMID- 9401080 TI - Malignant histiocytosis-like B-cell lymphoma, a distinct pathologic variant of intravascular lymphomatosis: a report of five cases and review of the literature. AB - Malignant histiocytosis (MH)-like B-cell lymphoma (BCL) is a neoplastic proliferation of large B cells clinically characterized by fever, hepatosplenomegaly, haemophagocytosis and abnormal laboratory data, without lymphadenopathy or skin lesions. Interestingly, most cases have been reported in Asian patients, and it is unclear whether MH-like BCL is biologically distinct from conventional large B-cell lymphomas. We report five Japanese patients with MH-like BCL. Biopsied specimens of bone marrow, liver and/or spleen showed infiltration of neoplastic B cells accompanied by haemophagocytosing histiocytes. Lymphoma cells were positive for CD19, CD20 and HLA-DR surface antigens, and negative for CD5 and CD10. In four cases elevated serum levels of interleukin (IL)-6 and the soluble IL-2 receptor isoform were noted, but not IL-1beta, IL-2 or tumour necrosis factor-alpha. Autopsies of two cases were pathologically diagnosed as intravascular lymphomatosis (IVL). Based on these observations, the current and nine previous cases reported as MH-like BCL in Japan were re evaluated. They appear to form a peculiar variant of IVL, characterized by bone marrow involvement at presentation, haemophagocytic syndrome, and a rapidly aggressive clinical course, but rarely neurological complications or skin lesions. This variant may merit separate consideration because of the problems posed in the initial diagnosis and therapeutic approaches. PMID- 9401081 TI - Analysis of p18INK4C in adult T-cell leukaemia and non-Hodgkin's lymphoma. AB - p18INK4C, a cyclin-dependent kinase inhibitor, is a homologue of p15INK4B and p16INK4A which are frequently altered in a variety of malignancies. We searched for structural alterations of the p18INK4C gene in 44 adult T-cell leukaemias (ATLs), 101 non-Hodgkin's lymphomas (NHLs), two polyclonal B-cell proliferations, seven ATL cell lines and seven leukaemia/lymphoma cell lines, by Southern blot and polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) analyses. No genomic alterations of the p18INK4C gene were found in any of the samples. By RT-PCR, p18INK4C was not expressed in three of five ATL cell lines, whereas it was expressed in all the non-ATL leukaemia/lymphoma cell lines. Tax did not inhibit the expression of p18INK4C in tax-expressing Jurkat cells. PMID- 9401082 TI - Bcl-6 gene hypermutations in diffuse large B-cell lymphoma of primary gastric origin. AB - The presence of the bcl-6 gene hypermutation was studied in 40 Hong Kong Chinese patients with diffuse large B-cell lymphoma. The primary sites of involvement were nodal in 18 cases and gastric in 22. Hypermutations at the E1.11 segment of bcl-6 gene were detectable in 16/22 (73%) primary gastric and 4/18 (22%) primary nodal lymphoma (P<0.01). Three of the 22 cases of primary gastric but none of the nodal lymphoma had mutations at E1.12. The proportion of hypermutated cases in the group with bcl-6 gene rearrangement was similar to that of the germ-line group (70% v 75%). High frequency of bcl-6 gene hypermutations was found in diffuse large B-cell lymphoma of the stomach and they were independent of rearrangement of the gene as detected by Southern analysis. Unlike gene rearrangement, hypermutations of the bcl-6 gene did not appear to carry any prognostic significance clinically. PMID- 9401083 TI - ALL R-87 protocol in the treatment of children with acute lymphoblastic leukaemia in early bone marrow relapse. AB - Seventy-three children with acute lymphoblastic leukaemia (ALL) in first bone marrow (BM) relapse, occurring within 30 months from complete remission (CR), were enrolled in an Italian cooperative study (ALL R-87 protocol). This treatment programme consisted of an induction phase with intermediate-dose cytarabine (IDARA-C) plus idarubicin (IDA) and prednisone (PDN), followed by a multidrug consolidation therapy and bone marrow transplant (BMT). 55/73 children achieved CR (75.3%); 15 (20.5%) failed to respond and three (4.2%) died during induction. The response rate was significantly higher for children with a first CR duration > or = 12 months (P=0.0005) and for those with a white blood cell (WBC) count at relapse < 20 x 10(9)/l (P=0.004). The estimated disease-free survival (DFS +/- SE) at 82 months was 0.18 +/- 0.05 for all responders, and 0.70 +/- 0.14 for allotransplanted patients versus 0.05 +/- 0.05 for those autografted (P=0.001). The estimated probabilities of survival +/- SE and event-free survival (EFS +/- SE) at 83 months were 0.16 +/- 0.07 and 0.13 +/- 0.04, respectively. for all enrolled children. Univariate analysis showed that age < 10 years at initial diagnosis and B-lineage immunophenotype favourably influenced both DFS (P=0.001) and EFS probabilities (P=0.0014 and P=0.012, respectively), whereas a first CR duration > or = 12 months and a WBC count at relapse < 20 x 10(9)/l were associated only with a better EFS rate (P=0.026 and P=0.004, respectively). Our results show the efficacy of the IDA plus IDARA-C schedule used in the ALL R-87 protocol in high-risk relapsed ALL children. Allogeneic BMT proved effective for patients with an HLA sibling donor. In a multivariate analysis, age > or = 10 years at initial diagnosis (P=0.016) and WBC count at relapse > or = 20 x 10(9)/l (P=0.048) were independently associated with a worse disease outcome. PMID- 9401084 TI - Granulocyte colony-stimulating factor given in addition to interferon-alpha to mobilize peripheral blood stem cells for autologous transplantation in chronic myeloid leukaemia. AB - In order to potentially mobilize and harvest the Ph cells observed in most patients with chronic myeloid leukaemia (CML) during interferon-alpha (IF-alpha) therapy, G-CSF (filgrastim), 5 microg/kg/d, was administered subcutaneously together with IF-alpha to 30 CML patients in haematological remission but with various degrees of cytogenetic remission, after IF-alpha therapy. Peripheral blood stem cells (PBSC) were harvested using standard aphereses from day 5 of G CSF Patients underwent one to four (median three) aphereses. Median total yields/kg were 7.6 (range 3.8-25) x 10(8) MNC, 3.4 (0-140) x 10(6) CD34+ cells, and 17 (1.1-107) x 10(4) CFU-GM. No patient had a significant increase in the percentage of Ph+ cells in the bone marrow under G-CSF therapy. The percentage of Ph+ cells in apheresis products tended to decrease between the first and the last apheresis (P = 0.05). 14 patients who were not responsive to IF-alpha were transplanted after conditioning with busulphan 16 mg/kg and melphalan 140 mg/m2. Median time to neutrophils > 0.5 x 10(9)/l was 20 d (16-114 d) and to platelets > 50 x 10(9)/l 18 d (12-149 d). Nine patients had a major cytogenetic response post graft, which correlated with the amount of Ph+ cells reinfused with the graft (P = 0.02). We conclude that this procedure is feasible, allowing the harvest of enough PBSC, some of them Ph- in patients who responded to IF-alpha, to allow autologous transplantation. PMID- 9401085 TI - Multiple myeloma: reduced plasma cell contamination in peripheral blood progenitor cell collections performed after repeated high-dose chemotherapy courses. AB - The possibility of reducing tumour cell contamination by cytotoxic drug courses prior to peripheral blood progenitor cell (PBPC) collection was evaluated in two consecutives groups of multiple myeloma (MM) patient candidates for autograft. All patients were at disease onset and received two VAD (vincristine, doxorubicin and dexamethasone) courses as initial debulking. In the first group (44 patients), mobilization and harvest were performed 'upfront', after a single cyclophosphamide (CY) administration of 4 g/m2; in the second group (17 patients), PBPC were collected at the end of a high-dose sequential chemotherapy programme, including: CY 5 g/m2, etoposide (VP16) 2 g/m2, a chemotherapy-free interval with three courses of high-dose dexamethasone, a final mobilizing CY at 7 g/m2. G-CSF was given following each high-dose cytotoxic drug. Cytofluorimetric analysis was performed to quantify progenitors (CD34+ cells) and plasma cells, identified by the high CD38 expression and/or CD38 and CD138 coexpression. Large amounts of PBPC were collected in either group (median harvested CD34+/kg: 15.8 x 10(6) and 13.4 x 10(6), respectively; P=0.9). Circulating plasma cells were significantly higher in patients mobilized 'upfront' compared to those who received the high-dose sequence (median peak values of CD38bright/microl: 39 and 10, respectively; P=0.02); a similar difference was observed in the amount of contaminating plasma cells in the harvest products (median CD38bright/kg: 7.4 x 10(6) and 1.3 x 10(6), respectively; P=0.02). The results demonstrate that an in vivo purging approach is feasible in myeloma patients through repeated high-dose chemotherapy courses; this may provide less-contaminated material suitable for further in vitro purging procedures. PMID- 9401086 TI - Reduction of heat-induced haemotoxicity in a hyperthermic purging protocol of murine acute myeloid leukaemic stem cells by AcSDKP. AB - The tetrapeptide AcSDKP (Goralatide) is a cytokine with known inhibitory effects on cell proliferation. Many purging agents used in autologous bone marrow transplantation protocols, including hyperthermia, preferentially kill cycling cells. A pretreatment with Goralatide offers a possibility to reduce the haemotoxicity in many purging settings. The impact of Goralatide on the hyperthermic purging protocol was investigated in normal and myeloid leukaemic (SA8) murine cells. The median survival time after transplantation (i.e. leukaemia incidences) was used as an in vivo parameter to determine the effects on leukaemic cells. The hyperthermic effect on normal and leukaemic cells was also investigated in vitro using the cobblestone area-forming cell (CAFC) assay. A heat treatment of 90 min at 43 degrees C resulted in a 4-log depletion of leukaemic stem cells. For normal progenitor cells (CFU-GM) a 2-log cell kill was shown. The reduction in proliferative activity of the CFU-GM after an 8 h incubation with 10(-9) M Goralatide resulted in a decrease in the heat sensitivity of the progenitor subset to approximately a 1-log cell kill. The leukaemic precursor cells seem insensitive to Goralatide inhibition, implicating an increase in the therapeutic window of the hyperthermic purging protocol. Finally, simulated remission bone marrow (5% leukaemic blasts) was incubated with Goralatide followed by a heat treatment of 90 min at 43 degrees C. Lethally irradiated (10 Gy) mice transplanted with heat-treated remission bone marrow (10(6) normal bone marrow cells versus 5 x 10(4) leukaemic cells) died of aplasia while Goralatide-pretreated remission bone marrow could rescue the irradiated mice without revealing leukaemic engraftment. These findings confirmed the enhanced protection against hyperthermia of the normal haemopoietic subsets by Goralatide and thus increased the success of the hyperthermic purging protocol. PMID- 9401087 TI - Antithymocyte globulin for patients with myelodysplastic syndrome. AB - Twenty-five transfusion-dependent myelodysplastic syndrome (MDS) patients (with < 20% blasts) were treated in a phase II study with antithymocyte globulin (ATG) at 40 mg/kg/d for four doses and then followed with blood counts every 2 weeks and clinic visits every 3 months, for a median of 14 months (range 1-38 months). 11 (44%) patients responded and became transfusion-independent after ATG, including three complete responses, six partial responses, and two minimal responses. Responses were observed in 9/14 patients (64%) with refractory anaemia (RA) and 2/6 patients (33%) with refractory anaemia with excess blasts (RAEB). Median response duration was 10 months (range 3-38 months). The Kaplan-Meier estimate of overall survival was 84% at 38 months, with one early death due to pneumonia and two deaths from disease progression to leukaemia. Side-effects consisted mainly of mild serum sickness in all patients. A single course of ATG restored haemopoiesis in some patients with MDS and was well tolerated. PMID- 9401088 TI - Evans syndrome complicating fludarabine treatment for advanced B-CLL. PMID- 9401089 TI - CHOP for recurrent TTP. PMID- 9401090 TI - The presence of the Ph-chromosome in different cell lineages in CML. PMID- 9401091 TI - Confirmation of frequent somatic deletion of the 13q12.3 locus encompassing BRCA2 in chronic lymphocytic leukaemia. PMID- 9401092 TI - Hepatitis G virus (HGV) and lymphoproliferative disorders. PMID- 9401093 TI - Trisomy 14 in association with myeloid malignancies. PMID- 9401094 TI - Conventional cytogenetics and fluorescence in situ hybridization (FISH) PMID- 9401095 TI - Activated protein C (APC) resistance: does it exist in Basques? PMID- 9401096 TI - [Intraoperative pain stimuli change somatosensory evoked potentials, but not auditory evoked potentials during isoflurane/nitrous oxide anesthesia]. AB - PURPOSE: Evoked potentials are used for intraoperative monitoring to assess changes of cerebral function. This prospective randomised study assesses the influence of surgical stimulation on midlatency components of somatosensory (SEPs) and auditory evoked potentials (AEPs) in anaesthetised patients. METHODS: After approval of the Ethics Committee and written informed consent 36 orthopaedic patients (34 +/- 15 y, 73 +/- 14 kg. 1.71 +/- 0.07 m, ASA I-II) were randomly included in the study. Anaesthesia was induced with 1.5 micrograms/kg fentanyl, 0.3 mg/kg etomidate and 0.1 mg/kg vecuronium. The lungs were intubated and patients normoventilated in steady state anaesthesia with isoflurane (end tidal 0.6%) and 66% nitrous oxide. 18 patients (group 1) were assigned to the SEP group: median nerve stimulation, recording at Erb, C 6 and the contralateral somatosensory cortex (N20, P25, N35) vs Fz. AEPs were recorded in group 2 (n = 18): binaural stimulation, recording at Cz versus linked mastoid (V, Na, Pa, Nb). Recordings were performed during 30 min before the start of surgery (baseline: BL), at skin incision (SURG1) and at the preparation of the periost (SURG2). Heart rate, mean arterial blood pressure, oxygen saturation, endtidal pCO2 and isoflurane (PetISO) concentrations were registered simultaneously. Data were analysed by one-way analysis of variance. Post hoc comparison were made by Mann Whitney U-Wilcoxon Rank Sum Test with p < 0.05 significant. RESULTS AND DISCUSSION: During steady state isoflurane anaesthesia surgical stimulation (SURG2) resulted in significant increases of N20 P25 amplitudes compared with BL (BL: 1.4 +/- 0.7 microV; SURG2: 2.0 +/- 0.8 microV; p < 0.05). Latencies of SEPs and midlatency components of AEPs did not change over time. There were no differences in autonomic parameters between SEP and AEP groups. MAP increased from 76 +/- 6 mmHg at BL to 93 +/- 16 mmHg at SURG1 and 96 +/- 17 mmHg at SURG2 (n = 36; p < 0.05). HR increased from BL (60 +/- 8 beats/min) to SURG2 (76 +/- 12 beats/min). Increases of amplitudes of midlatency SEP amplitudes indicate increased nociceptive signal transmission which is not blunted by isoflurane nitrous oxide anaesthesia. In contrast, unchanged AEPs indicate adequate levels of the hypnotic components of anaesthesia. PMID- 9401097 TI - Reduction of post-prandial motility by pinaverium bromide a calcium channel blocker acting selectively on the gastrointestinal tract in patients with irritable bowel syndrome. AB - BACKGROUND: Growing evidence points to irritable bowel syndrome physiologically as a disease of the enteric nervous system characterised by hypermotility. The aim of this study was to investigate the action of pinaverium bromide a calcium channel blocker acting selectively on the gastrointestinal tract on basal and post-prandial recto-anal motility of 40 irritable bowel syndrome patients in a random, double blind and placebo controlled trial. METHODS: Pinaverium bromide (50 mg) or placebo was taken orally t.i.d. with food. Myoelectrical and mechanical activities of the rectum and the internal anal sphincter were recorded before treatment for 2 h in the fasting state and for an additional 2 h post prandial. RESULTS: Post-prandial rectal spike amplitude and frequency as well as the spontaneous recto-anal inhibitory reflex frequency decreased after pinaverium bromide (P < 0.01) but not after placebo. CONCLUSIONS: These results suggest that the calcium channel blockers acting selectively on the gastrointestinal tract may have a therapeutic role in patients with irritable bowel syndrome. PMID- 9401098 TI - Syphilitic rectal ulcer associated with perforation of the hard palate: case report. AB - We report a case of a patient that presented with a perforated hard palate as a late complication due to an unsuspected syphilis. This disease first presented as a rectal ulcer which was misdiagnosed as an amebic proctitis. The patient received antiamebic treatment with a satisfactory outcome. He did not return for late control of the latter treatment and returned seeking medical advice six years later with the former complication. He tested positive for syphilis and appropriate treatment was performed. In addition, the ORL department recommended a palate prosthesis. PMID- 9401099 TI - Serum IgE level and clinical allergic diseases in the United Arab Emirates. AB - OBJECTIVE: To determine the serum IgE level in population and its association with allergic diseases in Al-Ain, United Arab Emirates (UAE). DESIGN: This is a prospective descriptive hospital-based study. SETTING: Al-Ain Medical District, Tawam Hospital, United Arab Emirates. SUBJECTS: Patients who were seen at Tawam Hospital for skin, allergic and respiratory diseases during 1996. RESULTS: The study was based on 228 patients. The sample consisted 95 (41.7%) males and 133 (58.3%) females. The mean and standard deviation of age were 25.89 +/- 15.87 years with range 1 and 72 years. The present study revealed that there were no statistically significant differences between serum IgE level and age groups, gender, clinical diagnosis and respiratory symptoms (p > 0.05). Of 59.2% of patients clinically diagnosed were possible IgE related. There was statistically significant differences between serum IgE level and cut (p < 0.030). There was a positive correlation between clinical diagnosis and cut. CONCLUSION: Consistencies and discrepancies between our findings and those from other studies with respect to allergic diseases and IgE level which support the hypothesis that allergic diseases are a multi-factorial diseases related to both familial and environmental influences. PMID- 9401100 TI - Floating Harbor syndrome. Case report and further syndrome delineation. AB - Floating Harbor syndrome was diagnosed in a 9 years old girl on the basis of short stature, delayed bone age, mild mental retardation, speech problems and specific craniofacial features. A detailed phenotype description is given and evaluated together with 18 other published case reports according to the concept of the Munich Dysmorphology Database (Stengel-Rutkowski et al., 1996) with the aim to delineate the spectrum of clinical and anthropological features. PMID- 9401102 TI - Characterization of two extreme variants involving the short arm of chromosome 22: are they identical? AB - The heteromorphic nature of the short-arms of human acrocentric chromosomes is considered the norm without any dire consequences. We characterized two highly unusual chromosome 22 variants with extremely enlarged short arms by routine and molecular cytogenetic techniques. Routine banding revealed that the two variants were not alike. Therefore, a characterization by fluorescent in situ hybridization (FISH) technique became warranted and revealed their remarkable differences. The first variant apparently had a tandem duplication of bands p11.2 ->p13, while the second variant had a loss of the beta-satellite and ribosomal DNA regions with an apparent amplification of the satellite III region. The formation of these extremely enlarged regions can occur by a variety of mechanisms whose clinical significance remains obscure. PMID- 9401101 TI - Systematic screening for fragile X syndrome in a cohort of 574 mentally retarded children. AB - In this study, we evaluated the prevalence of the fragile X syndrome in a cohort of 574 mentally retarded children. The only inclusion criterion was the diagnosis of mental retardation according to the DSM-IIIR classification. We used a PCR based strategy for the diagnosis of fragile X syndrome to facilitate systematic screening. This diagnostic scheme is based on an initial PCR to eliminate most fragile X-negative patients followed by Southern blotting for fragile X syndrome diagnosis. Altogether, 403 boys and 171 girls were tested. The prevalence of this genetic disorder was 1.9% (11/574) in the whole cohort and 2.5% (10/403) in boys. Only one case of fragile X syndrome was detected among the 171 girls tested (0.6%). Clinical examination, especially in the youngest children, was often unremarkable, and the only reason for suspecting fragile X syndrome was the presence of mental retardation. Thus, a systematic screening for the fragile X syndrome in mentally retarded children seems justified because of the importance of a precise diagnosis in genetic counseling. PMID- 9401103 TI - A recombination event close to HFE gene in hereditary hemochromatosis. AB - A candidate gene for Hereditary Hemochromatosis (HFE) has been recently cloned from a region 4 cM telomeric to HLA-A on the short arm of chromosome 6. This gene, defined HFE, is mutated in a high proportion of HFE patients. Positional cloning of HFE has been difficult, because of the extended region of linkage disequilibrium observed around the gene and of the rarity of recombination events in this DNA area. Here we describe a crossover in an Italian HFE patient which occurred close to the HFE gene in a restricted interval between D6S2221 and D6S2240-D6S2238 markers. The molecular analysis of this event and the segregation of the HFE mutations in the family are consistent with the position of the HFE gene telomeric to D6S2221. PMID- 9401104 TI - Clinical applications of primed in situ labelling (PRINS) rapid identification of a marker chromosome in a fetus. AB - Marker chromosomes pose a serious problem in clinical cytogenetic diagnosis since the conventional banding analyses are often not useful in identifying their origin or composition. In the absence of information, counseling as to the clinical significance and prognosis is difficult, especially in prenatal diagnosis. With the introduction of fluorescence in situ hybridization (FISH) marker identification has became feasible. However, FISH is relatively time consuming and expensive. In an effort to overcome these disadvantages, we have used primed in situ labelling (PRINS) technique as an alternative. Presented here is one case in which PRINS was useful in rapidly identifying the origin of a marker chromosome detected on amniotic fluid chromosome analysis. Based on our experience with this case and others, we propose that PRINS can become a viable and cost effective alternative to FISH and is as reliable as FISH in terms of accuracy, specificity, and sensitivity. PMID- 9401105 TI - 18p monosomy with midline defects and a de novo satellite identified by FISH. AB - We report a girl with an 18p deletion and showing a total GH deficiency, a single central maxillary incisor, and a pituitary dysplasia. This suggests that del(18)(p) could be involved in pituitary dysplasia. We review the association between midline developmental defects and chromosome 18 anomalies. This case is due to a de novo satellite resulting from an unbalanced translocation t(18p;13p) identified by FISH. This is the first case of this cytogenetic mechanism in the 18p monosomy. PMID- 9401106 TI - High recurrence of recombinants in a family with pericentric inversion of chromosome 18. AB - We report a family in which the father carries a pericentric inversion involving two third of the chromosome 18 (p11.2q22). Of his three children, the proposita and her youngest brother show partial duplication of the short arm and partial deficiency of the long arm; the oldest sister shows the other recombinant (partial duplication of the long arm and partial deficiency of the short arm). In the literature, we found only one family in which both recombinants of a parental pericentric inversion were present in the same offspring and none with three affected and both kinds of recombinants. A review of the reported familial cases reveals that the risk of aneusomy of recombination, at least for chromosome-18 inversion carriers, may be close to 20% and no only 5-10% as previously reported. PMID- 9401107 TI - Myotonic dystrophy protein kinase gene expression in skeletal muscle from congenitally affected infants. AB - Myotonic dystrophy (DM) is an autosomal dominant neuromuscular disorder characterized by marked variability of its clinical manifestations. The mutational basis of DM is an unstable (CTG)n trinucleotide repeat in the 3' untranslated region of the myotonic dystrophy protein kinase gene (DMPK). We used quantitative RT-PCR to determine DMPK mRNA levels in muscular biopsies from three congenitally affected (CDM) and two control infants. The CDM infants had increased DMPK mRNA levels, which were not correlated to increased expression of the mutant allele. This increase may be the consequence of a maturational muscular arrest, which may maintain an elevated level of DMPK mRNA until birth. PMID- 9401108 TI - A rare case of a liveborn with free, de novo and partial trisomy 12 and an unusual phenotype. AB - Individuals with free and total trisomy 12 are rare and always mosaic. Incidences of partial trisomy 12 are more frequent and are classified into trisomy 12p and trisomy 12q. The phenotype of both trisomy 12p and trisomy 12q is well described in the literature. We report here, the rare occurrence of a liveborn with free and de novo trisomy 12, albeit not the whole chromosome. The clinical description of this infant includes characteristics of trisomy 12p and trisomy 12q syndromes. Few additional anomalies present in the infant are unaccounted for by both syndromes. We anticipate that these characteristics are caused by trisomy 12q13, which to our knowledge has not been reported in a trisomy before. PMID- 9401109 TI - Mosaicism in trisomy 8: phenotype differences according to tissular repartition of normal and trisomic clones. AB - Three new observations of trisomy 8 mosaicism are presented. In two postnatal cases, both patients showed agenesis of corpus callosum associated with different clinical findings. In a third case, the prenatal diagnosis revealed trisomy 8 mosaicism exclusively in chorionic villi (CV) cells long term culture. Normal results were obtained in CV direct preparation and in cultured amniotic cells. In lymphocytes, the child showed low level trisomy 8 mosaicism. The only clinical findings were deep palmar and plantar furrows. The present cases as well as reports in the literature indicate that the variation in tissular repartition of normal and trisomic clones in trisomy 8 mosaicism is possibly responsible for the missing correlation between cytogenetic findings and clinical severity in this syndrome. PMID- 9401110 TI - CFTR gene analysis in cystic fibrosis patients: detection of 91% of molecular defects and identification of the novel mutation D979V. AB - More than 600 mutations have been identified in the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene and are known to cause cystic fibrosis (CF). This large number of mutations makes the search of the molecular defects in CF patients difficult. We have used the techniques of denaturing gradient gel electrophoresis (DGGE) and direct DNA sequencing to detect the mutations in 334 CF chromosomes mostly of French origin. The whole coding sequence of the CFTR gene corresponding to the 27 exons and their exon-intron boundaries was studied. 45 different mutations were identified. This method allowed us to detect the molecular defect in 90.5% of the mutant alleles and to report a novel mutation D979V. PMID- 9401111 TI - Detection of delta F508 mutation in cystic fibrosis patients from northwestern Mexico. PMID- 9401116 TI - [Lymphoplasmacellular osteomyelitis]. AB - Chronic lymphoplasmacellular osteomyelitis may occur in children, adolescents and adults, but has not been found in newborns or babies either in our series or in the literature. Symptoms suggesting an acute disease like fever are uncommon, but a primary chronic course with symptomatic and asymptomatic periods is typical. Pain and swelling are the main symptoms; painless masses are rare. In children and adolescents the clavicle and metaphyseal regions of long bones are typical sites of chronic abacterial osteomyelitis. In adults the clavicles or the first two ribs are mainly affected with synovitis of the adjacent joints, but the long bones are rarely involved. Laboratory findings are non-specific but important for the differential diagnosis. The sedimentation rate and c-reactive protein might be elevated. The X-ray examination shows osteolytic, sclerotic or mixed bony changes and, in case of a diaphyseal involvement, onionskinlike periosteal bone formation may be present, suggesting a malignant process. In late stages sclerotic bone formations may be seen as a rest. Uni- or multifocal lesions can be detected by bonescan, as can asymptomatic lesions. Magnetic resonance imaging shows gross signal intensity differences both in the bone and perifocal soft tissue and involvement of the synovium with gadolinium DPTA enhancement in T1 weighted images. In early stages of the disease granulocytes, microabscesses and new bone formations might suggest bacterial osteomyelitis that cannot be differentiated by histology. In intermediate phases lymphocytic and plasma cellular infiltrates are found, whereas in late phases sclerotic bone formations and fibrosis of the bone marrow are seen histologically. In chronic lymphoplasmacellular osteomyelitis, all clinical, radiological and histological findings, as well as negative bacteriological cultures, are mandatory and will allow a definitive diagnosis to be made. The disease may be uni- or multifocal, and new bone lesions may occur over time, as well as skin manifestations, which can be found years before or after bone involvement. The association with dermatological diseases and/or synovitis led to the acronym SAPHO syndrome (synovitis, acne, pustulosis, hyperostosis, osteomyelitis). For the treatment nonsteroidal anti-inflammatory drugs are effective for pain relief, reduction of swelling and dysfunction. Antibiotics have been used in several series and are not effective. Major surgery is not recommended even for recurrences, and the prognosis for growth and function is excellent in the long term despite recurrences over several years. PMID- 9401117 TI - New arguments to explain the high infection rate in posttraumatic spleenless patients. AB - To get more information about the high infection rate in splenectomized adult patients 211 spleenless patients were investigated with regard to clinical and laboratory data and compared to healthy blood donors. The results show that the infection rate is markedly increased to 30%. Splenectomized patients have decreased IgG levels which is due to diminished IgG1 and IgG4. Whereas IgA, complement factors C3, C4, and transferrin are not changed in patients without spleen, fibronectin and IgM are significantly reduced and the phagocytosis as well as the migration of neutrophilic granulocytes is impaired to 50%. With these changes in laboratory data it is possible to identify patients which bear an increased risk with regard to infection. PMID- 9401118 TI - Different roles of flowering-time genes in the activation of floral initiation genes in Arabidopsis. AB - We have analyzed double mutants that combine late-flowering mutations at four flowering-time loci (FVE, FPA, FWA, and FT) with mutations at the LEAFY (LFY), APETALA1 (AP1), and TERMINAL FLOWER1 (TFL1) loci involved in the floral initiation process (FLIP). Double mutants between ft-1 or fwa-1 and lfy-6 completely lack flowerlike structures, indicating that both FWA and FT act redundantly with LFY to control AP1. Moreover, the phenotypes of ft-1 ap1-1 and fwa-1 ap1-1 double mutants are reminiscent of the phenotype of ap1-1 cal-1 double mutants, suggesting that FWA and FT could also be involved in the control of other FLIP genes. Such extreme phenotypes were not observed in double mutants between fve-2 or fpa-1 and lfy-6 ap1-1. Each of these showed a phenotype similar to that of ap1-1 or lfy-6 mutants grown under noninductive photoperiods, suggesting a redundant interaction with FLIP genes. Finally, the phenotype of double mutants combining the late-flowering mutations with tfl1-2 were also consistent with the different roles of flowering-time genes. PMID- 9401119 TI - Genetic analysis of osmotic and cold stress signal transduction in Arabidopsis: interactions and convergence of abscisic acid-dependent and abscisic acid independent pathways. AB - To dissect genetically the complex network of osmotic and cold stress signaling, we constructed lines of Arabidopsis plants displaying bioluminescence in response to low temperature, drought, salinity, and the phytohormone abscisic acid (ABA). This was achieved by introducing into Arabidopsis plants a chimeric gene construct consisting of the firefly luciferase coding sequence (LUC) under the control of the stress-responsive RD29A promoter. LUC activity in the transgenic plants, as assessed by using in vivo luminescence imaging, faithfully reports the expression of the endogenous RD29A gene. A large number of cos (for constitutive expression of osmotically responsive genes), los (for low expression of osmotically responsive genes), and hos (for high expression of osmotically responsive genes) mutants were identified by using a high-throughput luminescence imaging system. The los and hos mutants were grouped into 14 classes according to defects in their responses to one or a combination of stress and ABA signals. Based on the classes of mutants recovered, we propose a model for stress signaling in higher plants. Contrary to the current belief that ABA-dependent and ABA-independent stress signaling pathways act in a parallel manner, our data reveal that these pathways cross-talk and converge to activate stress gene expression. PMID- 9401120 TI - The Arabidopsis deetiolated2 mutant is blocked early in brassinosteroid biosynthesis. AB - The Arabidopsis DEETIOLATED2 (DET2) gene has been cloned and shown to encode a protein that shares significant sequence identity with mammalian steroid 5 alpha reductases. Loss of DET2 function causes many defects in Arabidopsis development that can be rescued by the application of brassinolide; therefore, we propose that DET2 encodes a reductase that acts at the first step of the proposed biosynthetic pathway--in the conversion of campesterol to campestanol. Here, we used biochemical measurements and biological assays to determine the precise biochemical defect in det2 mutants. We show that DET2 actually acts at the second step in brassinolide biosynthesis in the 5 alpha-reduction of (24R)-24 methylcholest-4-en-3-one, which is further modified to form campestanol. In feeding experiments using 2H6-labeled campesterol, no significant level of 2H6 labeled campestanol was detected in det2, whereas the wild type accumulated substantial levels. Using gas chromatography-selected ion monitoring analysis, we show that several presumed null alleles of det2 accumulated only 8 to 15% of the wild-type levels of campestanol. Moreover, in det2 mutants, the endogenous levels of (24R)-24-methylcholest-4-en-3-one increased by threefold, whereas the levels of all other measured brassinosteroids accumulated to < 10% of wild-type levels. Exogenously applied biosynthetic intermediates of brassinolide were found to rescue both the dark- and light-grown defects of det2 mutants. Together, these results refine the original proposed pathway for brassinolide and indicate that mutations in DET2 block the second step in brassinosteroid biosynthesis. These results reinforce the utility of combining genetic and biochemical analyses to studies of biosynthetic pathways and strengthen the argument that brassinosteroids play an essential role in Arabidopsis development. PMID- 9401121 TI - Aux/IAA proteins repress expression of reporter genes containing natural and highly active synthetic auxin response elements. AB - A highly active synthetic auxin response element (AuxRE), referred to as DR5, was created by performing site-directed mutations in a natural composite AuxRE found in the soybean GH3 promoter. DR5 consisted of tandem direct repeats of 11 bp that included the auxin-responsive TGTCTC element. The DR5 AuxRE showed greater auxin responsiveness than a natural composite AuxRE and the GH3 promoter when assayed by transient expression in carrot protoplasts or in stably transformed Arabidopsis seedlings, and it provides a useful reporter gene for studying auxin responsive transcription in wild-type plants and mutants. An auxin response transcription factor, ARF1, bound with specificity to the DR5 AuxRE in vitro and interacted with Aux/IAA proteins in a yeast two-hybrid system. Cotransfection experiments with natural and synthetic AuxRE reporter genes and effector genes encoding Aux/IAA proteins showed that overexpression of Aux/IAA proteins in carrot protoplasts resulted in specific repression of TGTCTC AuxRE reporter gene expression. PMID- 9401122 TI - The adenylate cyclase gene MAC1 of Magnaporthe grisea controls appressorium formation and other aspects of growth and development. AB - Magnaporthe grisea, the causal agent of rice blast disease, differentiates a specialized infection structure called an appressorium that is crucial for host plant penetration. Previously, it was found that cAMP regulates appressorium formation. To further understand the cellular mechanisms involved in appressorium formation, we have cloned a gene (MAC1) encoding adenylate cyclase, a membrane bound enzyme that catalyzes the production of cAMP from ATP, by using a polymerase chain reaction-based strategy. The entire gene was isolated and subcloned from a large insert bacterial artificial chromosome library. Sequence characterization showed that MAC1 has a high degree of identity with other adenylate cyclase genes from several filamentous fungi as well as yeasts. Gene deletion resulted in reduced vegetative growth, conidiation, and conidial germination. Transformants lacking MAC1 were unable to form appressoria on an inductive surface and were unable to penetrate susceptible rice leaves. mac1- transformants were also sterile and produced no perithecia. Appressorium formation was restored in the presence of exogenous cAMP derivatives. These results confirm that cell signaling involving cAMP plays a central role in the development and pathogenicity of M. grisea. PMID- 9401123 TI - Antisense suppression of 4-coumarate:coenzyme A ligase activity in Arabidopsis leads to altered lignin subunit composition. AB - The phenylpropanoid enzyme 4-coumarate:coenzyme A ligase (4CL) is considered necessary to activate the hydroxycinnamic acids for the biosynthesis of the coniferyl and sinapyl alcohols subsequently polymerized into lignin. To clarify the role played by 4CL in the biosynthesis of the guaiacyl (G) and syringyl (S) units characteristic of angiosperm lignin, we generated 4CL antisense Arabidopsis lines having as low as 8% residual 4CL activity. The plants had decreases in thioglycolic acid-extractable lignin correlating with decreases in 4CL activity. Nitrobenzene oxidation of cell walls from bolting stems revealed a significant decrease in G units in 4CL-suppressed plants; however, levels of S lignin units were unchanged in even the most severely 4CL-suppressed plants. These effects led to a large decrease in the G/S ratio in these plants. Our results suggest that an uncharacterized metabolic route to sinapyl alcohol, which is independent of 4CL, may exist in Arabidopsis. They also demonstrate that repression of 4CL activity may provide an avenue to manipulate angiosperm lignin subunit composition in a predictable manner. PMID- 9401124 TI - EMF genes regulate Arabidopsis inflorescence development. AB - Mutations in EMBRYONIC FLOWER (EMF) genes EMF1 and EMF2 abolish rosette development, and the mutants produce either a much reduced inflorescence or a transformed flower. These mutant characteristics suggest a repressive effect of EMF activities on reproductive development. To investigate the role of EMF genes in regulating reproductive development, we studied the relationship between EMF genes and the genes regulating inflorescence and flower development. We found that APETALA1 and AGAMOUS promoters were activated in germinating emf seedlings, suggesting that these genes may normally be suppressed in wild-type seedlings in which EMF activities are high. The phenotype of double mutants combining emf1-2 and apetala1, apetala2, leafy1, apetala1 cauliflower, and terminal flower1 showed that emf1-2 is epistatic in all cases, suggesting that EMF genes act downstream from these genes in mediating the inflorescence-to-flower transition. Constitutive expression of LEAFY in weak emf1, but not emf2, mutants increased the severity of the emf phenotype, indicating an inhibition of EMF activity by LEAFY, as was deduced from double mutant analysis. These results suggest that a mechanism involving a reciprocal negative regulation between the EMF genes and the floral genes regulates Arabidopsis inflorescence development. PMID- 9401125 TI - A strong inhibitor of gene expression in the 5' untranslated region of the pollen specific LAT59 gene to tomato. AB - Promoter sequences that direct pollen-specific expression have been previously identified in the LAT59 (for late anther tomato) gene. Here, we show that the LAT59 sequences encoding the 5' untranslated region inhibit expression of reporter genes by > 20-fold in transient expression experiments and up to 300 fold after stable transformation. Inhibition occurred in somatic cells as well as in pollen. Our results indicate that the inhibitor still functions after pollen germination and therefore does not modulate the level of the LAT59 protein during pollen development. The presence of the leader sequence dramatically decreased mRNA accumulation but without affecting translation rate and mRNA stability. We believe that the leader inhibits transcription. We mapped the inhibitor to a region in the leader that coincides with a putative stem-loop and present evidence that this stem-loop participates in inhibition. PMID- 9401126 TI - Role of the sulfhydryl redox state and disulfide bonds in processing and assembly of 11S seed globulins. AB - Seed legumins contain two conserved disulfide bonds: an interchain bond (IE) connecting the acidic and basic chains and an intrachain bond (IA) internal to the acidic chain. Mutant subunits were constructed in which these disulfide bonds were disrupted. Oxidized glutathione stimulated the rate of assembly of trimers with unmodified prolegumin subunits. Stimulation was not detected during assembly of IE mutant subunits and was diminished for the IA mutant. Hexamer assembly with trimers of mature unmodified subunits required oxidizing conditions. Trimers composed of mature IE mutants did not form hexamers. Both mutant and non-mutant subunits accumulated in hexamers when the cDNAs were expressed in tobacco. Hexamer assembly in seeds probably involved trimers with a mixture of mutant and non-mutant subunits. Similarly, mixed trimers that were a mixture of mutant and non-mutant subunits assembled into hexamers in vitro. The results demonstrate the importance of disulfide bonds during the assembly of 11S globulins. PMID- 9401127 TI - Satellite RNA-mediated resistance to turnip crinkle virus in Arabidopsis involves a reduction in virus movement. AB - Satellite RNAs (sat-RNAs) are parasites of viruses that can mediate resistance to the helper virus. We previously showed that a sat-RNA (sat-RNA C) of turnip crinkle virus (TCV), which normally intensifies symptoms of TCV, is able to attenuate symptoms when TCV contains the coat protein (CP) of cardamine chlorotic fleck virus (TCV-CPCCFV). We have now determined that sat-RNA C also attenuates symptoms of TCV containing an alteration in the initiating AUG of the CP open reading frame (TCV-CPm). TCV-CPm, which is able to move systemically in both the TCV-susceptible ecotype Columbia (Col-0) and the TCV-resistant ecotype Dijon (Di 0), produced a reduced level of CP and no detectable virions in infected plants. Sat-RNA C reduced the accumulation of TCV-CPm by < 25% in protoplasts while reducing the level of TCV-CPm by 90 to 100% in uninoculated leaves of Col-0 and Di-0. Our results suggest that in the presence of a reduced level of a possibly altered CP, sat-RNA C reduces virus long-distance movement in a manner that is independent of the salicylic acid-dependent defense pathway. PMID- 9401128 TI - A functional S locus anther gene is not required for the self-incompatibility response in Brassica oleracea. AB - The self-incompatibility (SI) response in Brassica involves recognition of self pollen by the papillar cells of the stigma and is mediated by the products of genes localized at the S (self-incompatibility) locus. Two S locus genes, SRK and SLG, are thought to encode components of a receptor complex present in the female partner. The putative gene product of SLA, a third S locus-linked gene that is expressed specifically in anthers, is a candidate for the male component of the SI recognition system. The identification of a mutant SLA allele, interrupted by a large insert resembling a retrotransposon, in self-compatible Brassica napus initially suggested that SLA played an essential role in the SI response. In this study, we have characterized an SLA allele from a self-compatible B. oleracea var acephala line and show that it too is interrupted by a large insert. However, analysis of seven B. oleracea var botrytis lines exhibiting both self-compatible and self-incompatible phenotypes showed that these lines carry an S allele very similar or identical to that of the B. oleracea var acephala line and that the SLA gene is interrupted by an insert in all seven lines. The insertion of the putative retrotransposon was shown to interfere with gene expression, with no SLA transcripts being detected by RNA gel blot analysis in a self-incompatible B. oleracea var botrytis line carrying an interrupted SLA gene. These data indicate that a functional SLA gene is not required for the SI response in Brassica. PMID- 9401129 TI - A developmentally regulated MAP kinase activated by hydration in tobacco pollen. AB - A novel mitogen-activated protein (MAP) kinase signaling pathway has been identified in tobacco. This pathway is developmentally regulated during pollen maturation and is activated by hydration during pollen germination. Analysis of different stages of pollen development showed that transcriptional and translational induction of MAP kinase synthesis occurs at the mid-bicellular stage of pollen maturation. However, the MAP kinase is stored in an inactive form in the mature, dry pollen grain. Kinase activation is very rapid after hydration of the dry pollen, peaking at approximately 5 min and decreasing thereafter. Immunoprecipitation of the kinase activity by an anti-phosphotyrosine antibody is consistent with the activation of a MAP kinase. The kinetics of activation suggest that the MAP kinase plays a role in the activation of the pollen grain after hydration rather than in pollen tube growth. PMID- 9401130 TI - The impact of personality disorders on behavioral treatment outcome for social phobia. AB - The impact of personality disorders (PDs) on exposure in vivo treatment for social phobia was investigated in three groups of social phobics: social phobia without any PD (n = 30), social phobia with a single diagnosis of avoidant PD (n = 18) and social phobia with multiple PDs (n = 13). We hypothesized parallel change for social phobia with and without an avoidant PD with the latter group being more impaired before and after treatment. In order to test this hypothesis, confidence intervals for change were computed. In line with our hypothesis, social phobics in all three groups improved significantly during treatment and no interaction effects were found on the repeated MANOVAs. By using a confidence interval, parallel change was found on most measures. The impact of additional anxiety and mood disorders on treatment outcome was investigated separately. The analyses showed that an additional anxiety or mood disorder also did not predict outcome of exposure treatment. PMID- 9401131 TI - The perception of bodily sensations, with special reference to hypochondriasis. AB - Bodily sensations are relevant to problems such as hypochondriasis, but the issue of whether people are accurate in their perception remains unclear. The accuracy of perception of bodily sensations was analysed in 20 male and 20 female volunteers using two methods: a heart beat tracking procedure and the within-S correlational approach described by Steptoe and Vogele (1992, Behaviour Research and Therapy, 30, 597-607). The correlational approach involved monitoring of heart rate, skin conductance level and total respiratory resistance during relaxation and task periods, and computing correlations between appropriate physiological parameters and ratings of heart rate, sweaty hands and difficulty with breathing. In general, subjective ratings of bodily sensations were tied more closely with feelings of distress than with objective physiological state. Error scores on the heart beat tracking procedure showed no association with hypochondriacal concerns or with vigilant and avoidant coping styles measured with the Mainz Coping Inventory. Individuals varied considerably in accuracy as assessed with the correlational approach. However, there was a significant negative association between hypochondriacal concerns and accuracy of perception of sweat gland activity. The results are discussed in relation to measures of somatic perception and the experience of bodily sensations. PMID- 9401132 TI - Attentional biases for negative information in induced and naturally occurring dysphoria. AB - Two studies investigated the relationship between attentional biases for negative information and dysphoria--both induced (Study 1) and naturally occurring (Study 2). In a modified dot probe task a series of word pairs was presented, and Ss responded to probes that replaced one of the words in each pair. The stimuli included depression-related, anxiety-related and neutral words. To examine the time course of the attentional biases, there were three exposure durations of the word pairs: 14 ms (+ 186 ms mask); 500 ms and 1000 ms. In Study 1, the depressed mood induction procedure was associated with greater vigilance for depression related words at 500 ms, with a similar trend at 1000 ms. In Study 2, measures of depressed mood and vulnerability correlated positively with vigilance for negative words in the 1000 ms condition. There was no evidence from either study that depressed mood was associated with a pre-conscious bias for negative words (i.e. in the 14 ms masked exposure condition). However, this pre-conscious bias was associated with high trait anxiety in Study 2, consistent with previous research. The results are discussed in relation to theoretical and empirical work on cognitive biases in clinical and non-clinical anxiety and depression. PMID- 9401133 TI - Common childhood fears and their origins. AB - The present study examined rank orders and characteristics of childhood fears. A 'free option' approach ('What do you fear most?') deviated markedly from the fear rank order based on the Fear Survey Schedule for Children. A second aim of the study was to investigate the origins of prevalent childhood fears. In contrast to the results of Ollendick and King (1991, Behaviour Research and Therapy, 29, 117 123), conditioning was found to be the most commonly reported pathway related to exacerbation and onset of fears. Finally, special attention was given to the top intense fear in children, namely fear of spiders. Children who reported 'none', 'some' or 'a lot' of spider fear were compared with each other in terms of pathways. No differences between the three groups were found with respect to the frequency of modeling and information experiences. However, high fearful children more often reported conditioning experiences than low or moderate fearful children. PMID- 9401134 TI - Comparison of cognitive style in bulimia nervosa and depression. AB - This study investigated the global and specific cognitive style associated with bulimia nervosa. Three groups of women (women with bulimia nervosa, women with major depression, and controls) completed measures of eating disorder severity, depression, dysfunctional cognitions and irrational beliefs. The control group was found to report significantly lower levels of cognitive distortions and irrational beliefs overall than both women with bulimia nervosa and women with depression. However, no difference was found between the latter two groups. Furthermore, the pattern of individual cognitions and beliefs was exactly the same. When depression was statistically controlled, cognitive style no longer differentiated between the control group and two clinical groups. These results have implications for improving the effectiveness of cognitive behaviour therapy for bulimia nervosa. PMID- 9401135 TI - Disgust and disgust sensitivity in blood-injection-injury and spider phobia. AB - Blood-injection-injury (BII) phobics and spider phobics show markedly different cognitive, psychophysiological, and motoric reactions to activating stimuli. These observations have led theorists to question whether the emotion of fear mediates both phobias. The present study examined the role of disgust and disgust sensitivity in these subtypes of specific phobia. BII phobics, spider phobics, and nonphobics completed questionnaires and rated pictures of specific objects on fear and disgust scales. Questionnaire data indicated that phobic participants were higher than nonphobics on fear, and also on disgust sensitivity. The reaction of BII phobics to pictures of medical stimuli was one of disgust, rather than fear. The reaction of spider phobics to pictures of spiders was a combination of fear and disgust, though fear appeared to predominate. Results are discussed in view of current theories of emotional factors in specific phobia. PMID- 9401136 TI - Danger ideation reduction therapy (DIRT): preliminary findings with three obsessive-compulsive washers. AB - Three obsessive-compulsive patients received Danger Ideation Reduction Therapy (DIRT) in an initial treatment trial. All three subjects presented with contamination/washing concerns but refused to participate in exposure and response prevention. DIRT is solely directed at decreasing danger-related expectancies concerning contamination. DIRT procedures do not attempt to address inflated personal responsibility. In addition, DIRT does not involve direct or filmed exposure to contamination-related stimuli, or behavioural experiments. Components of DIRT include corrective information cognitive restructuring, filmed interviews, microbiological experiments, attentional focusing and Hoekstra's (1989) probability of catastrophe estimation task. Treatment consisted of between six and ten 1-hr weekly sessions. At post-treatment, substantial reductions in scores on the Padua Inventory, Maudsley Obsessional-Compulsive Inventory and two global rating scales were apparent for all subjects. These improvements were maintained at a 3-month follow-up. The theoretical and clinical implications of these preliminary findings are discussed. PMID- 9401137 TI - The relation between anxiety sensitivity and depression in children and adolescents referred for anxiety. AB - Research conducted with adult samples suggests that anxiety sensitivity is positively related to depression (Otto et al., 1995, Journal of Anxiety Disorders, 10, 117-123). The Childhood Anxiety Sensitivity Index (CASI, Silverman et al., 1991, Journal of Clinical Child Psychology, 20 162-168) was used in this study to provide an examination of the relation between anxiety, anxiety sensitivity, and depression in a sample of children and adolescents (N = 234) referred for anxiety disorders. A significant correlation between depression and anxiety sensitivity was found. This relation remained statistically significant when controlling for other aspects of anxiety (i.e. worry, physiological anxiety, and concentration). The similarities between these findings and findings obtained with adults are discussed, as well as suggestions for future research. PMID- 9401138 TI - The Reiss Screen for Maladaptive Behavior: confirmatory factor analysis. AB - We tested the factorial stability of the Reiss Screen for Maladaptive Behavior (Reiss, 1988a, The Reiss Screen for Maladaptive Behavior test manual). Reasonable fit was demonstrated in a geographically diverse sample of 448 individuals with mild, moderate, severe and profound mental retardation according to four measures of overall fit: RMSEA, ECVI, NNFI, and NFI. The results confirm Reiss' (1988a) factor solution of this widely used dual diagnosis (mental retardation and mental illness) screening instrument. PMID- 9401139 TI - A hospital attachment in restorative dentistry: a valuable but rare opportunity for general dental practitioners. PMID- 9401140 TI - Thiomersal sensitivity in health care workers. PMID- 9401141 TI - Tooth fairies. PMID- 9401142 TI - Factors influencing the diagnosis and management of periodontal disease by general dental practitioners. AB - OBJECTIVE: To identify factors influencing the diagnosis and management of periodontal disease by general dental practitioners. DESIGN: The study was conducted in two stages. 1. Analysis of returns to the Scottish Dental Practice Board. 2. Data collection via a postal questionnaire distributed to 500 general dental practitioners (27% of Scottish practitioners). RESULTS: 374 (75%) questionnaires were completed and returned. Analysis revealed the majority of treatment provided was in the form of simple scale and polishes, with multi-visit periodontal therapy comprising less than 0.2% of all non-surgical periodontal treatment. While the majority of respondents were confident in their ability to diagnose periodontal disease, only 40% were confident in treating it. Patient related factors were seen as the major hindrance to disease management, although time factors and the low level of fees were viewed by at least one half of respondents as major constraints. CONCLUSIONS: Greater efforts are required to motivate and inform patients of the necessity for and benefits of periodontal treatment, as is emphasis on appropriate treatment of early to moderate forms of disease. There is also a need for the development of appropriate, evidence-based clinical guidelines and fiscal arrangements to facilitate such provision. PMID- 9401143 TI - Are anterior occlusal radiographs still indicated for orthodontic assessment? AB - OBJECTIVE: This study examined the question of taking anterior occlusal radiographs (AO) in addition to a dental panoramic tomogram (DPT) using the Planmeca PM Proline CC. DESIGN: Retrospective single centre study. SETTING: Dental hospital radiology department. SUBJECTS: 100 pairs of DPTs and AOs which had been requested for orthodontic assessment were examined between April and July 1995. All DPTs were taken on a Planmeca PM Proline CC machine. RESULTS: The AO was assumed to be the gold standard and the diagnostic validity of the DPT can be assessed using values of sensitivity and specificity. 21% of DPTs were identified as requiring a supplementary radiograph to clarify the image of the anterior maxilla. CONCLUSIONS: A DPT machine with an adjustable focal trough has significantly reduced the need for supplementary AO radiographs. The AO is indicated when there is abnormality in the premaxilla or where dental malocclusion prevents ideal positioning of the patient in the machine. PMID- 9401144 TI - Prevalence of HCV infection in health care workers of a UK dental hospital. AB - OBJECTIVE: To determine the prevalence of hepatitis C virus (HCV) antibodies in a group of dental health care workers (DHCW). DESIGN: Retrospective cross sectional study. SETTING: A UK dental hospital. SUBJECTS AND METHODS: The sera of 167 unselected DHCW were tested for the presence of IgG antibodies to HCV using two, third-generation enzyme-linked immunosorbent assays (ELISA). HCV viremia was determined by reverse transcription polymerase chain reaction (RT-PCR) analysis. RESULTS: Two (1.2%) of the serum samples were found to be anti-HCV positive; one was also viremic. The two antibody-positive subjects were a qualified dental nurse and a student dental nurse, both females, without any known risk factor for HCV acquisition. No dentist was HCV seropositive. CONCLUSIONS: Since the prevalence of HCV infection in the UK general population varies between 0.08% and 0.55%, these results suggest that DHCW, and auxiliary staff in particularly, may have a slightly increased risk of HCV infection. PMID- 9401145 TI - The evaluation of SafeQuest--a computer-assisted learning program on cross infection control for the dental team. AB - OBJECTIVE: Evaluation of a computer-assisted learning (CAL) program (authored by a general dental practitioner-GDP) on cross-infection control (CIC) for the dental team. The aims of the evaluation were to determine how easy the program was to use, to determine if users found the information useful, to ensure that the content was accurate and to see what improvements could be made. METHODS: 96 questionnaires were distributed to dentists, 57 of whom responded (60%). 66% of the respondents were GDPs. RESULTS: Compared with earlier evaluations of hospital produced CAL programs, SafeQuest was equally well received. 96% of respondents found it easy to use and 93% gave a positive score for their overall impression. 64% felt that the program had extended their knowledge, despite the fact that 74% rated their knowledge on CIC as being good/excellent before using SafeQuest. Suggestions on how the content and presentation of the program could be improved led to changes to the final version. CONCLUSIONS: The SafeQuest program on cross infection control could be useful as a learning resource in general dental practice. The program, authored and produced by a GDP, compares favourably with those written in dental schools. PMID- 9401146 TI - A survey of dental professionals' health and well-being. AB - Past research has indicated that working within dentistry may offer a considerable threat to health. However, no data have previously been published regarding the general well-being and lifestyles of dental personnel. This descriptive study, which was undertaken by the BDA, aimed to examine the self perceived health and well-being of people working in dental surgeries and to describe their health related behaviours. PMID- 9401147 TI - Treatment of severe mental illness. PMID- 9401148 TI - My life on the street. AB - In this first-person account, the author chronicles her experience of being mentally ill and homeless in New York and New Jersey in the early 1980s. She describes surviving numerous traumatic events until she eventually realized her need for treatment and admitted herself to a mental institution. PMID- 9401149 TI - Families of borderline patients: a psychoeducational approach. AB - The development of the psychoeducational form of treatment described in this article has been prompted by changes in our understanding of borderline psychopathology and changes in the health care system in which these patients are treated. After reviewing these background changes, the authors describe the treatment itself, its form, its purpose, and the preliminary suggestions of its effectiveness. PMID- 9401150 TI - Using commercially available films to teach about borderline personality disorder. AB - Commercial films on videotape may be helpful in teaching medical students, residents, and other mental health trainees about various topics in psychiatry. Recommendations are made on the use of specific films, scenes, and characters to illustrate the DSM-IV diagnostic criteria for borderline personality disorder (BPD). The authors list 33 films depicting various aspects of BPD that might be used for teaching purposes. PMID- 9401151 TI - Aging and retirement: a difficult issue for individual psychoanalysts and organized psychoanalysis. AB - Organized psychoanalysis has a long history of discussing and formulating policies on retirement, but follow-through has been lax and few analysts retire unless forced by illness. Institutes tend to avoid and deny the problem of the impaired analyst. Procedural guidelines are needed for assessing competency and imposing involuntary retirement. The authors recommend that all institute appointments be time-limited and that medical clearance specifically addressing conditions potentially impairing professional functioning be required for appointment and renewal. PMID- 9401152 TI - The vision in supervision: transference-countertransference dynamics and disclosure in the supervision relationship. AB - The centrality of the supervision experience in the development of the supervisee's personal and professional capacities is addressed. The supervision relationship and process are explored in light of the potential effects of transference-countertransference configurations of supervisor and supervisee. Parallels between supervision and treatment are highlighted. The importance of developing and utilizing the capacity for reflectivity is reviewed, as is the impact of supervisee nondisclosure to supervisor. The direct use of countertransference experiences in the context of supervision is explored, and the centrality of self-disclosure is highlighted. It is recommended that supervisor and supervisee remain receptive to exploring these experiences in the service of developing a shared subjective sense of the patient, of increasing the supervisee's capacity to treat his or her patient, and of providing the supervisee with a novel, growth-enhancing relationship. PMID- 9401153 TI - The unbearable agony of being: interpreting tormented states of mind in the psychoanalysis of sexually traumatized patients. AB - This article focuses on the clinical importance of the disturbing transference countertransference matrix in the psychoanalysis of patients whose ego development was decisively influenced by early, traumatic sexual abuse. Dissociative defensive operations and "automatic" identifications are emphasized in accounting for the sadomasochistic and other characteristic features of the "traumatic" transference-countertransference ambiance. Two clinical vignettes depict the analyst's need to take his or her own disturbing experience as an object of analytic examination, while illustrating how "here-and-now" transference cues are used to interpret the patient's efforts to cope with overwhelming, traumatized states of mind. PMID- 9401154 TI - Methodological problems encountered in research with psychiatric inpatients: a case example. AB - In this article, problems associated with several methods commonly employed in research with psychiatric inpatients are discussed and the implications that these problems have for the validity of research with this population are explored, using an investigation of the relationship between moral reasoning and aggression among psychiatric inpatients as a case example. Specific issues examined include the adequacy of hospital records for diagnosing patients, the difficulty of determining when it is appropriate to approach recently admitted patients for research, and problems in the measurement of behavioral and psychological variables such as aggression and moral reasoning. Suggestions and recommendations for addressing these issues in future research are offered. PMID- 9401155 TI - Outbreak of Vibrio parahaemolyticus related to raw oysters in British Columbia. PMID- 9401156 TI - Outbreak of Salmonella enteritidis phage type 8 in a Montreal hotel. PMID- 9401157 TI - A case of human rabies contracted in Nigeria. PMID- 9401158 TI - Ciguatera fish poisoning linked to the ingestion of barracuda in a Montreal restaurant--Quebec. PMID- 9401159 TI - Transmission of hepatitis C virus infection associated with home infusion therapy for hemophilia. PMID- 9401160 TI - Ageism: the quiet epidemic. PMID- 9401161 TI - Age and depression in a nationally representative sample of Canadians: a preliminary look at the National Population Health Survey. AB - There are considerable inconsistencies in the literature concerning the relationship between age and depression. Recently, however, two independent studies in the U.S. have shown that the distribution is U-shaped with the lowest reported levels of depression at ages 45-49. Three reasons for past inconsistencies are identified and addressed using the 1994 National Population Health Survey by Statistics Canada. Using both a distress scale and a diagnostic measure, a substantially different relationship was found. The prevalence of distress decreased steadily with age until about 65, with only a slight increase afterwards for both males and females. After the introduction of several sociodemographic covariates, however, this relationship was clearly negative. These findings are discussed in terms of future research questions. PMID- 9401162 TI - Mortality and cognitive status among elderly Canadians living in the community and in institutions: the Canadian Study of Health and Aging. PMID- 9401163 TI - Predictors of dietary intake in Ontario seniors. AB - This study determined the independent association of 24 risk factors with dietary intake in community-living seniors. The study sample was 5,073 seniors for whom complete data were available from the 1990 Ontario Health Survey. Risk factors were items completed on an interviewer-administered health questionnaire. Diet Score, Mean Adequacy Ratio and energy were the diet outcomes derived from a self administered food frequency questionnaire. The independent association of risk factors with these diet outcomes was assessed with multiple linear regression analyses. Factors that were consistently and positively associated with diet outcomes included: education, income, social support, perceived health status, belief in the nutrition/health link, dependence in walking and vision. Factors that were consistently and negatively associated with diet outcomes included: chewing status, dentition, hearing, level of happiness and body mass index. These results provide a basis for the development of a screening tool for the identification of "at risk" subgroups of seniors. PMID- 9401164 TI - Nutrient intakes of senior women: balancing the low-fat message. AB - Nutrient intakes based on six days of food intake were collected from 52 elderly Nova Scotian women. Mean reported energy intake was 1600 kilocalories, of which fat contributed 31%. Mean intakes of zinc and Vitamin D were below recommendations. Other nutrients of concern were protein, calcium, folate, and vitamins B6 and B12. Nutrition education efforts should be directed at assisting older people to maintain the generally recommended low fat intake, while stressing the desirability of a balanced food intake of sufficient quantity, which includes low-fat milk and dairy products, lean meats and legumes as sources of nutrients in low supply. PMID- 9401165 TI - Canadian recommended nutrient intakes underestimate true energy requirements in middle-aged women. AB - To examine whether Health Canada's Recommended Nutrient Intakes (RNI) and FAO/WHO/UNU (Food and Agriculture Organization, World Health Organization, United Nations University) values provide accurate indices of true energy requirements, energy expenditure was determined using doubly labelled water (DLW) over 13 days in a group of 29 middle-aged women. Energy intakes were calculated from weighed food intake, and energy expenditures and intakes were then compared with individual calculated RNI requirements. The mean energy requirement as determined by DLW expenditure (9.56 +/- 0.53 MJ/d) was higher (p < 0.0001) than reported energy intake (7.08 +/- 0.30 MJ/d) and was higher (p < 0.004) than RNI mean energy requirement (7.97 +/- 0.18 MJ/d). The mean RNI for energy was also lower (p < 0.0001) than that derived from FAO data. These results suggest that current Health Canada RNIs are inadequate in predicting the energy needs of Canadian middle-aged women. PMID- 9401166 TI - Creation of priority criteria for corneal transplantation and analysis of factors associated with surgery following implementation. AB - PURPOSE: We sought to test the effectiveness and application of a system for prioritizing corneal disease patients for corneal transplantation. METHODS: All patients wait-listed for corneal transplantation in British Columbia in April 1995 were followed for 10 months to determine whether they received surgery and to assess the application of recently introduced priority criteria. RESULTS: The factors that determined whether a patient had surgery were as follows: having vision in one eye only, being female, having progressive disease, and experiencing pain. The surgeon involved was also a factor. Overall, the priority system did not adequately predict who had surgery and who did not have surgery. CONCLUSIONS: The priority system needs to be re-structured to reduce the contribution of months waited and to more adequately take into account patient need. Furthermore, its application by individual surgeons needs to be strengthened. PMID- 9401167 TI - Along the boardwalk: effects of a boardwalk on walking behaviour within a Nova Scotia community. PMID- 9401168 TI - Prostate cancer screening in the midst of controversy: Canadian men's knowledge, beliefs, utilization, and future intentions. AB - Despite controversy about prostate cancer screening, administrative data show that the use of prostate specific antigen (PSA) testing in Canada has increased. This study sought to determine awareness and knowledge of prostate cancer and screening, use to date, and future intentions to have a digital rectal examination (DRE) and PSA test among Canadian men aged 40 and over. Data were collected through a Canada-wide cross-sectional random digit dial telephone survey of 629 men. Awareness of DRE and PSA, use to date, and future intended use varied with age and education. Although only 9% of respondents had had PSA testing for screening, future intentions to undergo this test were higher than use to date. Knowledge of prostate cancer and screening controversies was low, and men received more information about PSA from the media than from doctors. Men would, therefore, benefit from age- and education-specific information regarding the factors to consider in making an informed choice about prostate cancer screening. PMID- 9401169 TI - A perspective on Canadian teenage births, 1992-94: older men and younger women? AB - This article uses vital statistics relating to births by Canadian mothers between 1992 and 1994 to examine the distribution of age of father by age of mother at the birth of the child. Over 77% of births to teenage mothers involved males who were older than the mother. At the time of birth of the child, the mean difference between age of the teenage mother and the father was 4.1 years, compared with a mean of 2.6 years for all mothers and fathers. For mothers below the age of 18 years, 37% of partners were within 2 years of the woman's age, 39% were 3 to 5 years older, and 24% were six or more years older. Family planning and sex education programs directed at the prevention of teenage pregnancy, especially if these programs are given in the elementary or high school system, would not necessarily reach older males, who make up the majority of partners in teenage pregnancies. PMID- 9401170 TI - Taking issue with alternative medicine label. PMID- 9401171 TI - Taking issue with alternative medicine label. PMID- 9401172 TI - Factors important in promoting mammography screening among Canadian women. AB - Among women aged 50 to 69 years, regular screening by mammography in combination with clinical examination, can substantially decrease the morbidity and mortality associated with breast cancer by facilitating early detection. Unfortunately, many Canadian women are not screened in accordance with current guidelines. Research to date is based primarily on large surveys conducted in the United States and less is known about the relevance of specific barriers to mammography screening among Canadian women. Multivariate results from the 1994-95 National Population Health Survey (NPHS) indicate that younger (40-49) and older (70+) women, those who are socioeconomically disadvantaged, and minority women are least likely to report having had a mammogram. Conversely, women with positive health behaviours, high social support, and positive mental health attributes are more likely to participate in mammography screening. These findings are discussed in terms of the implications for developing successful intervention programs for Canadian women and for setting priorities for further research. PMID- 9401173 TI - Impact of two cardiovascular disease reduction education programs varying in the type of nutrition information provided. AB - The purpose of the study was to determine the impact of two worksite cardiovascular nutrition education programs. Program 1 focused on information related to the skills needed to change dietary behaviours (1 session, 45 minutes). Program 2 focused on information related to skills as well as cardiovascular risk factors (1 session, 60 minutes). The study sample consisted of office employees at three worksites. The pretest consisted of questions pertaining to: frequency of consumption of high fat foods, knowledge related to the risk and skills components of the program, and self-report of family and personal history of cardiovascular disease. Of employees who completed the pretest, 67% (55/82) in Program 1, 88% (46/52) in Program 2, and 86% (30/35) in the control group completed the post-test (six weeks after the programs). The results of regression analysis indicated that participants of Program 1 (skills only) reduced their frequency of consumption of high fat foods (p < 0.01); no other variables were significant. Nutrition education programs for the prevention of cardiovascular disease should focus on information related to skills when limited time is available. PMID- 9401174 TI - Breast milk and the prevention of neonatal and preterm gastrointestinal disease states: a new perspective. AB - In this review of the protective properties of human breast milk, a new perspective is taken to underscore the passive and active protection properties of breast milk in providing specific protection against selective gastrointestinal disease affecting the neonate and preterm infant. The normal protective properties of the gastrointestinal epithelial barrier (immunologic and nonimmunologic) are considered as is the development of barriers to antigen absorption in the immature infant human intestine as a background for considering three accelerated gastrointestinal diseases-necrotizing enterocolitis, intestinal allergy, and bacterial gastroenteritis. In each of these conditions, the developmental protective defect is considered and the role of breast milk plays in filling the protective void discussed. Besides considering passive protection of breast milk including the new roles assigned to nutrients such as lactoferrin and nucleotides, and importance of active substances in breast milk such as growth factors, cytokines and hormones are discussed in the context of actively stimulating the infant's own intestinal defenses to function as a protective barrier. Future studies at the cellular and molecular level should be helpful in designing both preventative and treatment strategies to deal with these diseases. PMID- 9401175 TI - Growth factors in breast milk and their effect on gastrointestinal development. AB - Breast milk contains various biologically active factors including, hormones, peptide growth factors, and cytokines. Epidermal growth factor (EGF) and insulinlike growth factor-I (IGF-I) are two of the major milk-derived peptide growth factors. Colostrum contains higher levels of these growth factors than mature milk does, and, these factors are relatively resistant to proteolysis and stable in the G-I tract. There are specific receptors found in G-I mucosa. Luminal EGF and IGF-I stimulate growth and development of gastrointestinal tract. Intestinal epithelial Caco-2 cells are a good model for studying physiological roles of exogenous growth factors in the G-I development. Effect of EGF and IGF-I on proliferation, differentiation, and IGF-binding protein (IGFBP) production of intestinal Caco-2 cells were studied. Both EGF and IGF-I increased cell proliferation in dose dependent manner. The number of IGF-I receptors on Caco-2 cells increased after differentiation, in contrast to EGF binding which was reported to decrease. Caco-2 cells produced at least three IGFBPs, namely IGFBP 2, -3, and -4. The profile of these IGFBPs varied with differentiation. Secretion of IGFBP-2 and -3 increased with differentiation, but IGFBP-4 diminished. IGF-I stimulated mainly IGFBP-3 production, while EGF stimulated predominantly IGFBP-4. The effects of IGF-I and EGF on IGFBP secretion diminished with increasing cell differentiation. Thus, the interaction between intestinal epithelial cells and extrinsic growth factors are complex and the stage of differentiation is an important determinant of this phenomenon. PMID- 9401176 TI - Breastfeeding practices in Indonesia. AB - Breastfeeding promotion program was started by the paediatricians and others in 1977, and is becoming a strong activity since 1990 it was declared by the President of the Republic of Indonesia as a National Movement. One year later the First Lady stated the importance of every Indonesian mother to breast-feed her baby, and thereafter many hospitals created the so called "Baby Friendly Hospital". In this occasion we only limit with eminent topics, i.e., "Exclusive Breast-feeding" in Indonesia, and "Breastfeeding amongst Working Mothers." In fact, until now the percentage of mothers who breastfeed exclusively is very low. Although the ever breastfed babies in Indonesia is 97% (Kodyat, 1996) but the data of the "Exclusive Breastfeeding" of Indonesia is just like Pakistan and Thailand, i.e. nearly 2 months, whereas the Philippines and Ceylon showed a figure of 4 months, and India 5 months. The Home Health Survey (SKRT) data in 1992 showed that 63.7% of the babies were exclusively breast-fed until 3 months. Three quarter of the quality of the exclusive breastmilk is quite good, enough or excellent, whereas the other one quarter is poor and this should be interfered by increasing the quality of the breastmilk and/or adding other formula, to prevent the baby of getting "failure to thrive" (Suharyono, 1996). Working mothers use to do "Early Weaning Practices" with very high mixed feeding practices (Matulessy et al, 1996). Mothers have to go to work because they have to support their family income, but unfortunately most of them ignore their main task of care their children. IN CONCLUSION: the experience in Indonesia proves that a very hard work should still be continued on the effort of promoting breastfeeding, especially regarding the two issues, i.e. "Exclusive breastfeeding" (we do hope at least until 4 months) and the other issue is regarding the "Working Mothers". PMID- 9401177 TI - Blood nutrient indices in breast and formula fed infants: amino acids metabolic responses. AB - Human milk is generally considered as nutritionally the best food for infants, infant formula are usually manufactured to be as similar to human milk as possible, it may appear logical to attempt a metabolic response of formula fed infants similar to that of breast fed infants. We conducted a 8 week growth and metabolic study in 90 healthy term infants fed either human milk, whey dominant formula or casein dominant formula, each group consisted of 30 infants. Results revealed no significant differences were observed among the groups with respect to growth and anthropometric measurements. Serum total protein, albumin were not differ among breast-fed and formula fed infants. BUN and many plasma essential, amino acid were significant higher in formula-fed groups. While plasma tryptophan concentration was lower in two formula fed groups, in addition, plasma taurine was lower in casein dominant group at 4 weeks of age. These results suggested a reevaluation of protein quantity and quality in infant formula is needed. PMID- 9401178 TI - Perspective of a pediatric nephrology program: an 18 year retrospective. AB - We analyzed the patient profile in a pediatric nephrology training program, along with data collected over an 18 year period, to determine whether there is merit in the proposition that clinical training can be obtained equally well in internal medicine nephrology training programs. We also compared the rate of patient referral in an U.S. metropolitan area with a population of 1.2 million, in the first 9 years without the "gatekeeper" health insurance system and the next 9 years with managed care competition. Finally, we discussed guidelines for renal biopsy in the child and approaches to treatment as practiced in a pediatric nephrology program of almost two decades. We used the same NIH clinical data form throughout the 18 years of data collection to record clinical, laboratory and biopsy diagnosis, dialysis/ transplantation and other treatment data of patients entering our outpatient and inpatient services. Between 1977 and 1996, 3,150 new patients were examined for disorders related to the kidney. Twenty-one per cent of the patients were in the first year of life and 50% were younger than seven years of age. The majority of the 389 percutaneous renal biopsies were done in children under 10 years of age. In addition, almost half of the 112 pediatric dialysis/transplant patients presented before 10 years of age. Thus, the majority of patients were in the early years of life, with an unique pattern of renal diseases and issues regarding therapy which are clearly different from adulthood. Therefore we concluded that the existing data did not support the proposition that pediatric nephrology training be absorbed into internal medicine nephrology programs. The introduction of managed care competition did not affect the rate of patient enrollment. In fact, the rate of referrals in the latter 9 years paralleled the first 9 years. The factors which contribute to this outcome are discussed. Such data should be useful to those trying to meet the challenges of this competitive era. Finally, we discussed guidelines for renal biopsies in children and approaches to specific diseases. PMID- 9401179 TI - Bone fractures in children with chronic cholestasis. AB - The clinical features, biochemical data, roentgenographical bone changes and liver histological studies of four children with fractures related to chronic cholestasis are described. Two of them had Alagille syndrome and the other two, biliary atresia. Only one patient presented with forearm fracture associated with typical radiographic evidence of rickets. The others revealed generalized osteopenia together with fractures in arms, forearms, and wrists respectively. Of these, one with rickets had hypocalcemia and hypophosphatemia; one had decreased parathyroid hormone with mild hypocalcemia and hyperphosphatemia; one had hyperphosphatemia solely. Eucalcemia and euphosphatemia were noted in the remaining patient. Their bone lesions deteriorated in spite of treatment, except for the one with clinical rickets. All the patients succumbed eventually. It was concluded that the real pathogenesis of bone lesions in end-stage liver disease of chronic cholestatic children remains unclear. However, because rickets may intervene, continuing efforts to supply vitamin D and calcium since diagnosis of chronic cholestasis on a long-term basis, to prevent fracture, is still mandatory. PMID- 9401180 TI - Patch testing in the detection of cutaneous reactions caused by carbamazepine. AB - Carbamazepine is a widely used antiepileptic drug associated with various side effects including skin eruptions. Peroral provocation test with any suspected drug is a reliable method of investigating the etiology: however, it is both laborious and potentially dangerous to the patient. Patch testing has been reported with variable success in skin drug reactions. Four cases (one male, three females) with epileptic seizures were reviewed; all had received carbamazepine therapy with appropriate dosage, then suffered from various cutaneous reactions including maculopapular exanthema, exfoliative dermatitis, erythema multiforme and Stevens-Johnson syndrome after the initial therapy for two weeks to one month. Skin patch test was done with 1% and 10% carbamazepine in petrolatum applied on the back, then read at 48 and 72 hours. All four patients had positive allergic patch test reactions to carbamazepine. One patient had extreme (+3), one had strong (+2) and another two had weak (+) reactions. There were no any skin reaction to vehicle and control cases. This limited study demonstrates that patch testing may be useful in the detection or confirmation of any type of exanthematous eruption caused by carbamazepine. PMID- 9401181 TI - Retinopathy of prematurity in very-low-birthweight neonates: epidemiology and risk factors. AB - A retrospective study of 143 very-low-birthweight infants cared in a level III neonatal intensive care unit who had survived for at least 28 days. Initial eye ground evaluation was done at the postnatal age between 4 and 6 weeks. Follow-up evaluation was done every one to two weeks at the discretion of the ophthalmologists. Thirty-four variables were reviewed for each case. Statistical analysis was done for each variable, with the development of retinopathy of prematurity (ROP), severity of ROP and development of threshold ROP as the dependent variables, by Mann-Whitney U test or X2 test when adequate. Variables with P-valu < 0.05 were included in multiple regression. One hundred and thirty eight cases were survived for more than 28 days with their eyes been checked. Twenty-six (18.8%) of them developed ROP. The prevalence of stage I was 2.2% (3/138), stage II was 3.6% (5/138), stage III was 12.3% (17/138), and stage V was 0.7% (1/138). Threshold disease, stage 3 (+) and above, was found in 5 cases (3.6%). Seventeen variables were found to be correlated with the development of ROP. Only the duration of continuous positive airway pressure (CPAP) was significantly correlated to the development of ROP in multivariate logistic regression. Fifteen variables were correlated with the severity of ROP, but only peak direct bilirublin level, peak total bilirubin level and duration of CPAP could entered multiple stepwise linear regression. Thirteen variables were correlated with the development of threshold ROP, but only episodes of septicemia enter the multivariate logistic regression. We postulate that the longer duration of CPAP in ROP cases may reflect the severity of apnea and episodes of hypoxic attacks. Reducing episodes of apnea may prevent the development of ROP. The number of episodes of septicemia was the only significant variable for threshold ROP so that infection control is important for the prevention of threshold disease. PMID- 9401182 TI - Analysis of total IgE and allergen-specific IgE antibody levels of allergic children in Taiwan. AB - With advances in technology, several in vitro screening tests such as MAST and CAP system have been used for analyzing the allergens involved in allergic diseases such as bronchial asthma (BA), allergic rhinitis (AR), atopic dermatitis (AD) and urticaria. In this study, CAP system (Pharmacia, Sweden) was used to screen the prevalence of allergens responsible for these atopic diseases. A total of 392 children were enrolled in this study retrospectively, all these atopic children visited the allergy clinic of the Department of Pediatrics, National Taiwan University Hospital during the period March 1995 and August 1995. Our results showed: (1) Among these 392 allergic children, included 82 BA, 70 AR, 22 AD, 156 BA + AR. 8 BA + AD, 12 AR + AD, and 42 AD + AR + AD: (2) House dust mites (Dermatophagoides pteronyssinus: D. p and Dermatophagoides farinae: D. f) are the most common allergens triggering atopic disease in the Taiwan area. (3) Total IgE level is the highest in three combined allergic disease (BA + AR + AD) [2179.9 +/ 504.2KU/L] and lowest in single disease (AR) [503.1 +/- 84.8 KU/L]. Mite specific IgE (D. p + D. f-specific IgE) concentration is also the highest in three combined disease (BA + AR + AD) [499.1 +/- 86.0KU/L] and lowest in AR [159.5 +/- 47.5 KU/L], (4) elevated specific IgE antibody to egg white and milk were found in 68.4% and 47.4% of patients with AD and/or urticaria. In conclusion, these data suggest that house dust mites, are the most important allergens in respiratory allergy as well as in atopic dermatitis, while food allergens play relatively important roles only in skin allergy. Furthermore, the highest IgE level was noted in children with combined allergic diseases. PMID- 9401183 TI - Myocarditis with complete atrioventricular block associated with herpes simplex virus infection: report of one case. AB - Myocarditis with complete atrioventricular block is a very unusual complication of the herpex simplex infection. We report a 10-year-old boy infected very likely by the herpes simplex virus and who presented with high fever, erythema multiforme, complete atrioventricular block, and Adams-Stokes seizures. Emergent temporary pacemaker was performed for bradycardia. A sixteen-fold rise in herpes simplex antibody titer by a complement fixation method occurred within two weeks. Normal cardiac rhythm recovered 11 days later with a sequela of complete right bundle branch block after 2 years follow-up. PMID- 9401184 TI - DiGeorge syndrome with microdeletion of chromosome 22q11.2: report of one case. AB - DiGeorge syndrome (DGS) is a congenital anomaly involving developmental defects of the third and fourth pharyngeal pouches. Thymic aplasia or hypoplasia, parathyroid aplasia or hypoplasia, cardiac malformations, and dysmorphic facies are characteristics features. We present a case which had thymic aplasia, hypocalcemia, facial dysmorphism (hypertelorism, low set ears, cleft of soft palate, fish-like mouth and micrognathia) and congenital heart disease (ventricular septal defect, perimembranous type). The T-cell immunologic functions as a percentage of T-cell and phytohemagglutinin stimulation test were within normal range matched with age. Molecular study showed microdeletion of chromosome 22q11.2 by genotype analysis, but chromosome study of high-resolution cytogenetic analysis by G-banding technique was normal. To our knowledge, about 90% of DiGeorge syndrome patients show chromosome abnormalities, most involving chromosome 22 (monosomy of 22q11.2). In the past, most cases were proven by high resolution cytogenetic analysis or fluorescence in situ hybridization(FISH). We report a case of DGS in Taiwan with microdeletion of chromosome 22q11.2 detected by genotype analysis. PMID- 9401185 TI - A giant retroperitoneal ganglioneuroma with intraspinal involvement: report of one case. AB - A 9-year-old girl was discovered to have had a huge retroperitoneal mass causing moderate hydronephrosis and anterior displacement of the right kidney and which was found on a renal ultrasonographic examination. A series of examinations including (1)IVP, (2) abdominal computed tomography, (3) MRI and(4) incisional biopsy revealed a giant dumb-bell shaped retroperitoneal ganglioneuroma with intraspinal involvement. Secondary scoliosis was also noticed. We successfully resected the huge ganglioneuroma of the right retroperitoneum in a two stage procedure: her postoperative course was uneventful. PMID- 9401186 TI - Congenital short bowel syndrome with left acheiria: report of one case. AB - A case of congenital short bowel syndrome with left acheiria, hemivertebra and dextrocardia is described. Dilatation of the fetal bowel was observed at the 24th week of gestation during a routine ultrasonic scan of a healthy 23-year-old primigravida from a non-consanguineous marriage. Amniocentesis and chorionic villus sampling were denied. The baby was delivered at 38 weeks of gestational age. After delivery, multiple anomalies were noted: left acheiria, congenital short bowel syndrome (15 cm in length of the ileum), pseudo-obstruction of intestine, dextrocardia, and hemivertebrae. We suspected these abnormalities might be due to a vascular accident or failure of lateralization during the early gestational period. To our knowledge, these combinations have not been reported previously in English literature. PMID- 9401187 TI - Aplasia cutis congenita: report of one case. AB - We present a female newborn with a skin defect on the vertex. Aplasia cutis congenita was diagnosed. No associated anomaly was found. There was no family or drug history noted. At one-year follow-up, the lesion tended to contract and became epithelialized. We report this case and also review the literature, and then we suggest a checklist for aplasia cutis congenita. PMID- 9401188 TI - Meningococcal disease in students at the University of Southampton: update. PMID- 9401189 TI - Uptake of influenza vaccine in high risk patients. PMID- 9401190 TI - Detecting vancomycin intermediate Staphylococcus aureus. PMID- 9401191 TI - Converting hospital computer systems. PMID- 9401192 TI - Health care quality leaders address patient confidentiality. PMID- 9401193 TI - Wireless, infrared "tag" system improves patient care. PMID- 9401194 TI - A challenge for nursing education. AB - Computer technology has the potential to help nursing programs to establish conditions that allow for academic success of special needs students without compromising the caliber of instruction or academic standards. As the capabilities of computers have increased in the last decade, a variety of exciting new tools have been created that have the potential to enhance the success of students with special needs. Effective instructional methods must be developed and implemented to make use of the power provided by these tools. Limited research is available in the nursing and education literature to support use of computer adaptations for students with special needs. There needs to be initiation of research based study of the tools and proposed adaptations. Educators, administrators, and researchers need to collaborate in this effort to transform the potential of technology into reality. PMID- 9401195 TI - Presentation hardware options. PMID- 9401196 TI - Criteria for nursing information systems as a component of the electronic patient record. An international Delphi study. AB - In many countries nurses lack an adequate tool to assist in determining the essentials of an information policy in health care institutions and to outline the nursing component of the electronic patient record. In the United States criteria exist for systems that support the nursing process and for the electronic patient record, and the United Kingdom has the disposal of an Information Management and Technology Strategy that includes nursing information. The objective of this study was to determine international criteria for nursing information systems when such systems become part of the electronic patient record. Using the Delphi methodology, criteria for nursing information systems development, content, structure, and use are established by an international panel of 36 experts in three succeeding rounds. Most criteria gained consensus and are useful for application in practice for development of information policy and information systems for nursing. Eventually, the list of criteria will be included in a nursing information reference model. Nursing will benefit from the application of the reference model and the criteria to develop adequate information and communication technology. PMID- 9401197 TI - Nursing the community in cyberspace. AB - As nursing expands more into the community, the role of health promotion is finding its way on the information superhighway. Through NetWellness, nursing faculty experts at the University of Cincinnati College of Nursing and Health are providing health care information via the internet. Not only is the world wide web a site of their nursing practice, but also faculty are identifying new linkages to add to the College's curriculum. PMID- 9401198 TI - Learning environment in information technology. The views of student nurses. PMID- 9401199 TI - Validation by school nurses of the Nursing Intervention Classification for computer software. AB - Validation of standardized nursing language for use by specialty nurses is important for the design of computer software. The purposes of this study were to validate the usefulness of the 433 interventions in the Nursing Intervention Classification (NIC) for school nurses and to identify interventions that could be omitted from computer software for school nurses. A school nursing listserv, SCHLRN-L, was used to recruit volunteers. Ninety-three volunteers from the listserv also recruited 26 school nurses who were not members of the listserv. The total sample was 102 school nurses from 25 states and other areas, 76 listserv volunteers, and 26 others. E-mail was used to send and receive the survey forms to portions of the sample. A majority of interventions (n = 241; 56%) were selected as used by more than 50% of the sample. Of these, 53 direct care interventions were selected as used by more than 80% of the sample. Fifty interventions were not used by 100% of the sample. E-mail was a useful means to obtain a national sample and collect data. PMID- 9401200 TI - Functions of SH2 and SH3 domains. PMID- 9401201 TI - Function of PTB domains. PMID- 9401202 TI - Pleckstrin homology domains. PMID- 9401203 TI - Structure and function of LIM domains. PMID- 9401204 TI - WW (WWP) domains: from structure to function. PMID- 9401205 TI - Modular domains of focal adhesion-associated proteins. PMID- 9401206 TI - Physiological function of receptor-SH2 interactions. PMID- 9401207 TI - The IRS-signaling system: a network of docking proteins that mediate insulin and cytokine action. PMID- 9401208 TI - PDZ domains and the formation of protein networks at the plasma membrane. PMID- 9401209 TI - Mechanism and function of signaling by the TGF beta superfamily. PMID- 9401210 TI - Notch receptors, partners and regulators: from conserved domains to powerful functions. PMID- 9401211 TI - Signaling through Grb2/Ash-control of the Ras pathway and cytoskeleton. PMID- 9401212 TI - Genetic analysis of sevenless tyrosine kinase signaling in Drosophila. PMID- 9401213 TI - An overview of thyroid carcinoma. AB - Approximately, 12,000 cases of thyroid carcinoma are diagnosed each year. This disease entity accounts for 1.5 percent of all malignancies and is clinically evident in four percent to seven percent of the United States population. This paper provides an overview of thyroid carcinoma, including types, incidence, predisposing factors, evaluation, and treatment. PMID- 9401215 TI - The duPont Hospital for Children: a brief history. PMID- 9401214 TI - Predicting red blood cell transfusions in very low birth weight infants based on clinical risk factors. AB - OBJECTIVE: To describe the clinical factors most predictive of red blood cell transfusion in very low birth weight (VLBW) infants. STUDY DESIGN: Retrospective review of VLBW infants cared for at a single level III NICU during a two year period, n = 199. RESULTS: Overall transfusion requirement was 4.6 +/- 6.2 transfusions/infant/hospital course. Length of hospital stay, days of mechanical ventilation, requirement for dopamine support, birth weight, initial hematocrit, periventricular leukomalacia and necrotizing enterocolitis all independently correlated with number of transfusions and donors. Bronchopulmonary dysplasia and patent ductus arteriosus were associated with donor but not transfusion number. CONCLUSIONS: Our data characterize the population of VLBW infants with the greatest blood transfusion and donor requirement. Further investigation is needed to target this population for interventions to reduce blood exposure. PMID- 9401216 TI - The perfect physician. PMID- 9401217 TI - Surface-marker cluster design criteria for 3-D bone movement reconstruction. AB - When three-dimensional (3-D) human or animal movement is recorded using a photogrammetric system, bone-embedded frame positions and orientations are estimated from reconstructed surface marker trajectories using either nonoptimal or optimal algorithms. The effectiveness of these mathematical procedures in accommodating for both photogrammetric errors and skin movement artifacts depends on the number of markers associated with a given bone as well as on the size and shape characteristics of the relevant cluster. One objective of this paper deals with the identification of marker-cluster design criteria aimed at the minimization of error propagation from marker coordinates to bone-embedded frame position and orientation. Findings allow for the quantitative estimation of these errors for any given cluster configuration and suggest the following main design criteria. A cluster made up of four markers represents a good practical compromise. Planar clusters are acceptable, provided in quasi-isotropic distribution. The root mean square distance of the markers from their centroid should be greater than ten times the standard deviation of the marker position error. The second objective of this paper deals with the identification of the optimal cluster position and orientation on the limb aimed at the minimization of error propagation to anatomical landmark laboratory coordinates. Cluster position should be selected to minimize skin movement artifacts. The longest principal axis of the marker distribution should be oriented toward the relevant anatomical landmark position. PMID- 9401218 TI - Joint kinematics simulation from medical imaging data. AB - A method for joint kinematics simulation is described. Kinematics parameters are determined from the relative displacement of marker sets placed on anatomical landmarks of surface models generated from medical imaging contour data. The landmarks are identified manually on fingers in multiple positions. A mathematical algorithm was then used to ascertain the kinematics axes of motion of the fingers. Once these axes are located, they are used as the base of a real time interactive simulation of the finger. The entire simulation was accomplished in a high-resolution graphics environment. A full complement of interactive tools (virtual dials and buttons controlled via mouse) was used to enhance the user interface. The development of the system, the model and the advantages and disadvantages of the method are discussed. PMID- 9401219 TI - Simultaneous and nonlinear identification of mechanical and reflex properties of human elbow joint muscles. AB - The naturally coexisting intrinsic mechanical and reflex properties of the human elbow joint were identified simultaneously using nonlinear, time-delay, continuous-time, and dynamic models. Angular random perturbations of small amplitude and low bandwidth were applied to the joint using a computer-controlled servomotor, while the subject maintained various levels of mean background muscle torque. Joint neuromuscular dynamics were identified from the measured elbow angle and torque. Stretch reflexes were modeled nonlinearly with both dynamic and static reflex gains. A continuous-time system identification method was developed to estimate parameters of the nonlinear models directly from sampled data while retaining realistic physical or physiological interpretations. Results from six subjects showed that dynamic stretch reflex gains, joint stiffness, and viscosity generally increased with mean background muscle torque; and that dynamic stretch reflex gain was higher during muscle stretch than that during muscle shortening. More importantly, the study provided realistic simultaneous estimates of the relative contributions of intrinsic mechanical and reflex actions to net joint torque. In particular, reflexively-mediated stiffness generated a significant portion of the total joint stiffness and the percentage varied systematically with background muscle torque. PMID- 9401220 TI - An advanced demultiplexing system for physiological stimulation. AB - A CMOS very large scale integration (VLSI) chip has been designed and built to implement a scheme developed for multiplexing/demultiplexing the signals required to operate an intracortical stimulating electrode array. Because the use of radio telemetry in a proposed system utilizing this chip may impose limits upon the rate of data transmission to the chip, the scheme described herein was used to reduce the amount of digital information which must be sent to control a large quantity (up to several hundred) of stimulating electrodes. By incorporating multiple current sources on chip, many channels may be stimulated simultaneously. By incorporating on-chip timers, control over pulse timing is assigned to the chip, reducing by up to fourfold the amount of control data which must be sent. By incorporating on-chip RAM, information associated with the desired stimulus amplitude and pulse timing can be stored on chip. In this manner, it is necessary to send control information to the chip only when the information changes, rather than at the stimulus repeat rate for each channel. This further reduces the data rate by a factor of five to ten times or more. The architecture described here, implemented as an eight-channel stimulator, is scalable to a 625-channel stimulator while keeping data transmission rates under 2 Mbps. PMID- 9401221 TI - Noninvasive glucose sensing utilizing a digital closed-loop polarimetric approach. AB - A polarimetric glucose sensor utilizing a digital closed-loop controller was designed and implemented during this study. Its potential as a noninvasive glucose sensor was evaluated in vitro for both glucose-doped water and bovine aqueous humor mediums. A physiological hyperglycemic concentration range was used in both calibration and validation of each set of experiments. Ideally, the end application of this system could estimate blood glucose concentrations indirectly by measuring the amount of rotation of a light beam's polarization state after it propagates through the aqueous humor contained within the anterior chamber of the eye. The polarimeter designed in this study differs from similar investigated systems in that it utilizes a digital closed-loop control system. This type of controller was implemented in order to further improve system repeatability and stability without sacrificing accuracy. Unique to this investigation, independent validation sets other than those used to create each respective calibration model were obtained. The results of the glucose-doped water experiments yielded mean standard errors of prediction for calibration and validation of 6.91 and 8.84 mg/dl, respectively. The mean standard errors of prediction during calibration and validation of the glucose-doped aqueous humor experiments were higher at 27.20 and 27.47 mg/dl, respectively, due to medium degradation over time while exposed to air. PMID- 9401222 TI - Real-time multichannel computerized electrogastrograph. AB - The purpose of this study was to develop a real-time multichannel computerized electrogastrograph (EGG) to measure and analyze electrical signals from the human abdominal surface. A soft-contact matrix composed of 25 cutaneous electrodes is embedded evenly in a latex mat. The mat can be firmly attached to the abdominal surface by drawing a vacuum between the matrix and the stomach. Twenty-five high amplification filter/amplifiers provide a high signal-to-noise ratio and flat amplitude response for a signal between 0.02 and 0.12 Hz (1.2-7.2 cpm). The computer program provides waveform and frequency analysis for any chosen channel and mapping analyses for all 25 channels. A two-dimensional propagation exploration program was also developed. Using four different mapping analysis program subroutines, the optimal points for analyzing the EGG signals can be reliably found and variability of these locations can be observed easily. Results show differences in the EGG mappings of normal and abnormal subjects. PMID- 9401223 TI - An integral equation model for intracardiac electrogram sensing. AB - Electrogram sensed by an intracardiac electrode has long been characterized based on two approaches: 1) presume that the electrode is very small and does not disturb the potential prior to applying the electrode, and 2) take an average of the prior potential over the electrode surface. In fact, any intracardiac sensing electrode has a finite surface area where electrical charges are induced and disturb the external potential field, thus, the sensed potential is different from the potential prior to placing the electrode. In this paper, an integral equation model is proposed based on the current continuity equation in homogeneous myocardial medium. The new model can accurately characterize the electrogram sensed by an electrode with a non-negligible surface area and a load impedance. The new model can be solved numerically via the method of moments to obtain the potential induced on the electrode surface by an arbitrary dipole volume source. As an application of the proposed theory, several electrode configurations with different loads have been analyzed with an intent to show that a finite electrode surface will significantly reduce the electrogram peak amplitude and slope, and a load impedance lower than 20 k omega will also degrade the electrogram sensitivity. PMID- 9401224 TI - An efficient algorithm for computing multishell spherical volume conductor models in EEG dipole source localization. AB - Computationally localizing electrical current sources of the electroencephalographic signal requires a volume conductor model which relates theoretical scalp potentials to the dipolar source located within the modeled brain. The commonly used multishell spherical model provides this source potential relationship using a sum of infinite series whose computation is difficult. This paper provides a closed-form approximation to this sum based on an optimal fitting to the weights of the Legendre polynomials. The second-order (third-order) approximation algorithm, implemented by a provided C-routine, requires only 100 (140) floating point operations to compute a single scalp potential in response to an arbitrary current dipole located within a four-shell spherical volume conductor model. This cost of computation represents only 6.3% (8.9%) of that required by the direct method. The relative mean square error, measured by using 20,000 random dipoles distributed within the modeled brain, in only 0.29% (0.066%). PMID- 9401225 TI - ECG coding by wavelet-based linear prediction. AB - This paper presents a novel coding scheme for electrocardiogram (ECG). Following beat delineation, the periods of the beats are normalized by multirate processing. After amplitude normalization, discrete wavelet transform is applied to each beat. Due to the period and amplitude normalization, the wavelet transform coefficients bear a high correlation across beats at identical locations. To increase the compression ratio, the residual sequence obtained after linear prediction of the significant wavelet coefficients is transmitted to the decoder. The difference between the actual period and the mean beat period, and that between the actual scale factor and the average amplitude scale factor are also transmitted for each beat. At the decoder, the inverse wavelet transform is computed from the reconstructed wavelet transform coefficients. The original amplitude and period of each beat are then recovered. The approximation achieved, at an average rate of 180 b/s, is of high quality. We have evaluated the normalized maximum amplitude error and its position in each cycle, in addition to the normalized root mean square error. The significant feature of the proposed technique is that, while the error is nearly uniform throughout the cycle, the diagnostically crucial QRS region is kept free of maximal reconstruction error. PMID- 9401226 TI - A noninvasive technique for detecting obstructive and central sleep apnea. AB - A new noninvasive method to detect obstructive and central sleep apnea [(OSA) and (CSA)] events is described. Data were collected from ten volunteer subjects with a previous diagnosis of OSA while they were titrated for continuous positive airway pressure (CPAP) therapy. Apneic events were identify by analyzing of estimated airway impedance determined from pressure and airflow signals delivered from CPAP. To enhance performance of this technique, a single-frequency (5 Hz with 0.5 cmH2O peak-to-peak amplitude) probing signal was superimposed on the applied CPAP pressure. The results indicated that estimated airway impedance during OSA (mean: 17.9, SD: 3.4, N = 50) was significantly higher then during CSA (mean: 4.1, SD: 1.7, N = 50). When the estimated impedance of OSA and CSA events were compared to a fixed threshold, 100% of all events can be correctly categorized. These results indicate that it may be possible to diagnose OSA and CSA noninvasively based upon this technique. The instrument and the algorithm required are relatively simple and can be incorporated in a home-based device. If this method was used for prescreening apnea patients, it could reduce cost, waiting time, and discomfort associated with traditional diagnostic procedures. PMID- 9401227 TI - Separation of discontinuous adventitious sounds from vesicular sounds using a wavelet-based filter. AB - The separation of pathological discontinuous adventitious sounds (DAS) from vesicular sounds (VS) is of great importance to the analysis of lung sounds, since DAS are related to certain pulmonary pathologies. An automated way of revealing the diagnostic character of DAS by isolating them from VS, based on their nonstationarity, is presented in this paper. The proposed algorithm combines multiresolution analysis with hard thresholding in order to compose a wavelet transform-based stationary-nonstationary filter (WTST-NST). Applying the WTST-NST filter to fine/coarse crackles and squawks, selected from three lung sound databases, the coherent structure of DAS is revealed and they are separated from VS. When compared to other separation tools, the WTST-NST filter performed more accurately, objectively, and with lower computational cost. Due to its simple implementation it can easily be used in clinical medicine. PMID- 9401228 TI - Simulations of the behavior of synaptically driven neurons via time-invariant circuit models. AB - This paper describes computer simulations of the behavior of Hodgkin-Huxley neurons, based on a redefinition of membrane and synaptic connections as time invariant circuit elements. Examples are given showing that this self-consistent equivalent circuit representation allows very efficient computer simulations and could facilitate the introduction of detailed biological neurons into formal neural networks. PMID- 9401229 TI - Reliability of percent distribution of power of the electrogastrogram in recognizing gastric electrical uncoupling. AB - The aim of this study was to examine the reliability of percent distribution of electrogastrographic (EGG) power in recognizing gastric electrical uncoupling. Sixteen anaesthetized dogs underwent laparotomy and implantation of six pairs of stainless-steel wire electrodes. Distal stomach was measured and three sections with approximately equal lengths were defined. Two pairs of electrodes were implanted in each section. Eight-channel EGG was also recorded. Three separate half-hour recordings were made: in the basal state; after a full circumferential separation of the distal antral section from the rest; after a second circumferential cut completely separating the middle from the proximal sections. EGG digital power spectra were split into three frequency ranges and dynamics of percent distribution of power was statistically examined. After the first cut, changes in the percent distribution of EGG power in the normal range were not significant (p = 0.2). Significant changes in the low range were noted (p < 0.05) and changes in the high range were borderline nonsignificant (p = 0.056). After the second cut, changes in percent distribution in the normal and the high range became significant (p < 0.01) while changes in the low range were insignificant (p = 0.075). Severe uncoupling was reflected in EGG by significant changes in the high-frequency range without internal tachygastria necessarily being present. PMID- 9401230 TI - Changing trend of Salmonella infection. PMID- 9401231 TI - Oxidants, antioxidants and diseases--a brief review. AB - Although oxygen (O2) is needed to the body, partially reduced forms of O2 and some of their derivatives collectively called reactive oxygen species (ROS), are highly toxic prooxidants to the body. To prevent ROS toxicity body's endogenous and exogenous antioxidants (AOS) act in concert. To maintain health a balance between pro and anti oxidants is very necessary. PMID- 9401232 TI - Evaluation of transperineal template implant technique in Indian cervical carcinoma patients. AB - The prognosis in advanced cervical cancer patients is poor specially in presence of distorted anatomy, gross residual growth etc. In these cases template implant offers good option for treatment. We have carried out the procedure in 19 patients with acceptable level of complication. Preliminary results have been described. PMID- 9401233 TI - Anti-neutrophil cytoplasmic antibody (ANCA)--a review. PMID- 9401234 TI - Clinton to apologize for tests. PMID- 9401235 TI - Costly medicines only for staff--patient sues NDMC hospital for Rs. 25,000. PMID- 9401236 TI - Diagnosis and monitoring of thyroid diseases. PMID- 9401237 TI - UV-dependent melanocyte plasticity--the structure-function relationship. AB - The UV response of marginal melanocytes in vitiliginous skin was studied using a whole skin organ culture technique. This method assesses the plasticity of melanocytes in response to UV. It is observed that there is a sequential increase in catecholoxidase production and in the volume and dendricity of the melanocytes on exposure to a single pulse of UV, reaching a peak at 3 1/2 h. From this study it is evident that the melanocyte shows a prominent structural and functional plasticity in response to UV. Implicit is the utilisation and transduction of UV energy by the melanocyte, for transcriptional and translational activity, enhancing catecholoxidase and cell structural protein production. PMID- 9401238 TI - Prevalence of bacterial prostatitis in benign prostatic hyperplasia. AB - To define the prevalence of bacterial prostatitis and urinary tract infection (U.T.I) among benign prostatic hyperplasia patients (B.H.P) undergoing prostatectomy trans urethral resection of prostate (T.U.R.P.), 100 consecutive patient has their preoperative and post-operative urine cultures along with tissue culture of the resected prostatic tissue. Our data suggests that significant incidence (42%) of bacterial growth in prostatic tissue-occurs in patients with B.H.P. Pre-existing U.T.I. is not a reliable indicator by which this group could be identified pre-operatively and prostatic infection could be treated. PMID- 9401240 TI - Characterisation of Vibrio cholerae 0139 isolated from diarrhoeal stools. AB - A total of 174 samples of acute diarrhoeal stools received over a period of seven months, yielded 101 isolated morphologically and biochemically resembling Vibro cholerae. Serologically, 57(56.4 per cent) of the 101 isolates were identified as V. cholerae 01 and remaining 44 (43.6 per cent) as V. cholerae 0139. Keeping in mind the unique potential of 0139 among non 01 vibrio to cause epidemics, we decided to undertake the study of biochemical characters and physiological behaviour of all the 44 V. cholerae 0139 isolates. All the stains were Voges Proskauer's test positive' haemagglutinating and grew in the presence of 6 per cent sodium chloride. 13 (29.5 per cent) strains showed haemolytic activity. Nine (20.5 per cent) were polymyxin-B sensitive and 4 (9.0 per cent) fermented lactose. All the isolates showed considerable degree of homogeneity in their biochemical and physiological properties, some characters define them to be closer to El Tor biotype. PMID- 9401239 TI - Anaerobic and aerobic microflora of pouch of Douglas aspirate v/s high vaginal swab in cases of pelvic inflammatory disease. AB - The anaerobic and aerobic bacterial flora in pouch of Douglas (POD) aspirate and high vaginal swabs (HVS) was studied in 43 cases of pelvic inflammatory disease (PID) using standard techniques. High vaginal swabs from 20 healthy women were included as controls. Anaerobic and aerobic bacteria were isolated from 37 (86%) HVS and 31 (72%) POD aspirates from these 43 women. A total of 100 aerobic and 10 anaerobic bacterial strains were recovered from both the sites. Coagulase negative staphylococci (28), Escherichia coli (23) and Streptococcus faecalis (14) were predominant aerobic bacteria. Of the anaerobes, peptostreptococci species and Bacteroides species were more common, polymicrobial flora (more than one type of organism) was present in a total of 27 specimens. However mixture of anaerobic and aerobic bacteria were seen in only 5 specimens. Of the 20 control specimens, ten were positive for organisms. Ten aerobic and 3 anaerobic bacterial strains were recovered. PMID- 9401241 TI - Study of pathogenicity markers of staphylococci isolated from clinical specimens. AB - A total of 165 strains of coagulase positive staphylococci (CPS) and 39 strains of coagulase negative Staphylococci (CNS) isolated from various clinical specimens studied for biochemical and enzyme profile showed overlapping of the key characters of pathogenicity. Anaerobic mannitol fermentation (69.20%), phosphatase (58.97%) and penicillinase (58.97%) production was remarkable amongst CNS. Both CPS and CNS showed increased resistance to penicillin and other antimicrobials. Besides increased frequency of isolation of CNS from pathological specimens, they elaborate singly or in combination, the recognized virulence factors. PMID- 9401242 TI - Pulmonary infection caused by Mycobacteria other than M. tuberculosis in and around Calcutta. AB - A study was undertaken to determine the prevalence rate of atypical mycobacteria in indolent fibrocavitary pulmonary diseases. 450 sputum specimens were examined and cultured, of which 15 were identified as atypical mycobacteria on repeated culture. No previous study was recorded from Calcutta. Compared to the results of previous workers from different regions of India, the prevalence rate seems high in this part of the country. PMID- 9401243 TI - Seroprevalence of syphilis by TPHA test. AB - A total of 1074 sera obtained from clinically suspected cases of Syphilis and various risk groups were screened for antitreponemal antibodies by Treponema pallidum Haemagglutination (TPHA) test and cardiolipin antibodies by the Venereal Disease Research Laboratory (VDRL) tests. The TPHA test was positive in 291 (27.09) per cent. The TPHA was positive in 53.31 per cent (161 out of 302) patients of primary syphilis, 54.17 per cent (26 out of 48) of secondary, 32.76 per cent (19 out of 58) of tertiary syphilis, 17.12 per cent (19 out of 111) of other Sexually Transmitted Disease (STD) patients, 25.32 per cent (60 out of 237) pregnant women and 1.89 per cent (6 out of 318) healthy individuals. TPHA was found to be superior to VDRL test in all the study groups. Almost a total agreement was seen between the TPHA test and VDRL test with a titre of 1 in 16 & above; while in as many as 32.39 per cent sera with VDRL titre of 1 in 8 TPHA was negative indicating that the VDRL titres above 1 in 8 should be considered as true reactives to minimize the biological false positive reactions. PMID- 9401244 TI - Trends of HIV infection in the blood donors of Delhi. AB - Screening of HIV Infection was made mandatory for every unit of blood collected for transfusion in Delhi, India since 1989. Ten Zonal Blood Testing Centres have been identified which test all the blood collected for HIV by 29 blood blanks for the city. Reports from these testing centres have been analysed yearwise to find out the magnitude and trends of HIV infection in different groups of blood donors. Although initially there was no difference in HIV Sero-reactivity in different blood donors categories (between 1 & 2 per 1000 blood donors samples tested) but subsequently there is significant increase (5.24/1000 in 1992 & 7.48/1000 in 1993) in the HIV sero-reactivity in replacement donor category possibly because professional donors donate blood in the guise of being replacement donors. The fact which comes out clearly is that HIV infection is present in all sections of the population in Delhi and mandatory HIV Screening of all blood collected for transfusion is justified. PMID- 9401245 TI - Coronary artery-intramyocardial sinusoid communication in a case of pulmonary atresia with intact ventricular septum. AB - Intramyocardial sinusoid--coronary artery fistulous communications are well established channels in pulmonary atresia with intact ventricular septum. Angiographically flow can be demonstrated from the right ventricular cavity into the coronary arteries. We have histologically demonstrated such a communication in a case of pulmonary atresia with intact septum. PMID- 9401246 TI - Hepatoblastoma with complete regeneration of liver and with metastasis to lungs and brain within six months after partial hepatectomy--a case report. PMID- 9401247 TI - Post-transplantation epididymitis associated with cytomegalovirus. AB - Cytomegalovirus (CMV) infection, though usually systemic, has been known to cause localised involvement of organs like lung, liver, testis and gastrointestinal tract. We report a case of cytomegaloviral infection involving the epididymis without systemic manifestations in an young male one month after renal transplantation. The diagnosis was made on histopathologic examination of the epididymo-orchidectomy specimen. Clinical improvement occurred after the emergency epididymo-orchidectomy. To the best of our knowledge, only three cases of CMV epididymitis have been described in the world literature--two in transplant patients and one in an AIDS patient. PMID- 9401248 TI - Echinococcosis involving the breast: diagnosis by fine needle aspiration cytology. PMID- 9401249 TI - Cytologic diagnosis of neurilemomas of male breast by fine needle aspiration. AB - Among 217 cases of male breast lesions subjected to fine needle aspiration cytology(FNAC) over a period of nine years, three cases were found to be having neurilemomas, The review of literature reveals the first case of neurilemoma of male breast diagnosed by FNAC was reported in 1992. Very scanty literature is available on this rare tumour of male breast. The aspirates yielded cellular smears composed of clusters of spindle shaped cells and Verocay bodies. Histopathological examination of the excised masses confirmed the cytological diagnosis. PMID- 9401250 TI - Childhood cancer: where do we stand? PMID- 9401251 TI - Effect of zinc supplementation on cell-mediated immunity and lymphocyte subsets in preschool children. AB - OBJECTIVE: In a zinc supplementation trial (with a significant impact on diarrheal morbidity), to evaluate effect of zinc supplementation on cellular immune status before and after 120 days of supplementation. DESIGN: A double blind, randomized controlled trial with immune assessment at baseline and after 120 days on supplement. SETTING: Community based study in an urban slum population. SUBJECTS: Randomly selected children (zinc 38, control 48), had a Multitest CMI skin test at both times. In 66 children (zinc 22, control 34), proportions of CD3, CD4, CD8, CD16, CD20 cells and the CD/CD8 ratio were also estimated using a whole blood lysis method and flowcytometry. INTERVENTION: Zinc gluconate to provide elemental zinc 10 mg daily and 20 mg during diarrhea. MAIN OUTCOME RESULTS: Regarding CMI, the percentage of anergic or hypoergic children (using induration score) decreased from 67% to 47% in the zinc group, while in the control group it remained unchanged (73% vs 71%) (p = 0.05). The percentage of children deteriorating between first and second tests was significantly lower in the zinc group (13% vs 33%, p = 0.03). Regarding lymphocyte subsets, the zinc group had a significantly higher rise in the geometric means of CD3 (25%, p = 0.02), CD4 (64% p = 0.001), and CD4/CD8 ratio (73% p = 0.004) with no difference in CD8 and CD20. The rise in CD4 was significantly higher in the zinc as compared to the control group; the ratio of geometric means was 1.45 (95% CI, 1.03-2.01). CONCLUSION: Zinc supplementation improves cellular immune status, which may have been one of the mechanisms for observed impact of zinc supplementation on diarrheal morbidity. PMID- 9401252 TI - Prevalence of malnutrition and intestinal parasites in preschool slum children in Lucknow. AB - OBJECTIVE: To assess the point prevalence of intestinal parasites and their association with nutritional parameters. SETTING: Anganwadi centers under the Integrated Child Development Scheme (ICDS) in Lucknow, North India. DESIGN: Cross sectional survey. METHODS: By random draw, 32 out of 153 Anganwadi centers were selected. All eligible subjects registered with the Anganwadi worker were enrolled. These were 1061 children (48.3% girls and 51.7% boys) between the ages of 1.5 to 3.5 years. RESULTS: Of these, 67.6% were underweight (weight for age < 2 SD), 62.8% were stunted (height for age < -2 SD) and 26.5% were wasted (weight for height < -2 SD). Parasites were detected in 17.5% (95% CI 15.3%-19.9%) children by a single direct fecal smear examination. Of these, Ascaris lumbricoides was found in 124 (68.1%) and Giardia lamblia in 60 (32.9%). There was no association between weight or height and parasite positivity. The mean hemoglobin levels for children who were smear positive versus smear negative for ascaris or giardia were 9.1 g/dl and 9.6 g/dl, respectively (p < 0.0001). CONCLUSION: In the urban slums the point prevalence of intestinal parasites is 17.5% in the preschool children. Malnutrition and low hemoglobin levels are also widely prevalent. Urgent remedial steps are needed on community basis to improve their nutritional status and control parasitic infestation. PMID- 9401253 TI - Cardiovascular responses to treadmill exercise testing in anemia. AB - OBJECTIVE: To study exercise performance on a treadmill in anemic children. DESIGN: Prospective case control study. SETTING: Department of Pediatrics and Intensive Care Unit, Department of Medicine, King George's Medical College, Lucknow. SUBJECTS: The study population consisted of 41 cases of anemia (10 mild, 21 moderate and 10 severe) and 11 normal age and height matched children aged between 7-12 years. METHODS: These subjects were exercise tested on Quinton Model Q5000 treadmill using Modified Naughton Q5000 protocol. Heart rate, systolic blood pressure, double product, ECG changes, exercise duration and metabolic equivalents achieved during peak exercise were studied. Statistical analysis was performed using analysis of variance (ANOVA) test. RESULTS: No significant difference was observed in values of resting heart rate, heart rate at peak exercise, recovery heart rate, blood pressure response, resting double product, double product at peak exercise, recovery double product and ECG changes in any of the study groups (p > 0.05). However, the gain in heart rate at peak exercise compared to basal value, and double product, total exercise duration and metabolic equivalent (MET) values at peak exercise were significantly low in anemic children on comparison to normal controls (p < 0.001). CONCLUSIONS: Cardiovascular responses are blunted in anemia, mainly because of depleted cardiac reserve. PMID- 9401254 TI - Alkali therapy for neonates: where does it stand today? PMID- 9401255 TI - Measles control in India: additional immunization strategies. PMID- 9401257 TI - Clinical profile of persistent diarrhea in a DTTU. PMID- 9401256 TI - Prevalence of underweight, stunting and wasting. PMID- 9401258 TI - Maternal abortions and birth of Down syndrome offspring. PMID- 9401259 TI - Propionic acidemia in the newborn. PMID- 9401260 TI - Valvular heart disease: rheumatic or rheumatoid? PMID- 9401261 TI - Congenital chylothorax. PMID- 9401262 TI - Tuberculosis verrucosa cutis. PMID- 9401263 TI - Age for typhoid vaccination. PMID- 9401264 TI - Aluminium phosphide poisoning: a growing concern in pediatric population. PMID- 9401265 TI - Neonatal skin lesions. PMID- 9401266 TI - A cheap alternative to a stadiometer. PMID- 9401268 TI - Consumer Protection Act and the pediatrician. PMID- 9401267 TI - Safe bilirubin level for term babies with non-hemolytic jaundice. PMID- 9401269 TI - Placenta: an alternative source of hematopoietic stem cells. PMID- 9401270 TI - Antimalarial vaccine. PMID- 9401271 TI - Sunlight exposure in young infants. PMID- 9401272 TI - HTLV-1-associated diseases. AB - HTLV-1-infection is associated with a variety of human diseases including adult T cell leukemia (ATL) and non-neoplastic inflammatory diseases. The latter includes HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) and HTLV-1 uveitis (HU) and other diseases with unestablished associations such as arthropathy, pneumopathy, dermatitis, exocrinopathy and myositis. ATL is defined as neoplastic clonal growth of HTLV-1-infected T-cells and is characterized by unique clinical features including hypercalcemia and severe organ infiltration of leukemic cells. HAM/TSP and HU are characterized by infiltration of HTLV-1 infected lymphocytes and dysregulated production of cytokines. Four major subtypes (genotypes) of HTLV-1 have been identified, which depend more on geography than disease. No disease-specific variants or mutations have been identified to date. A viral transcriptional regulator, Tax, regulates virus and cellular gene expression through binding to transcription factors or other cytoplasmic cellular molecules. Aberrant expression of cellular genes will affect fundamental cellular functions. The interaction between HTLV-1-infected cells and different kinds of cells in the host appears to be one of the basic mechanisms underlying the development of HTLV-1-associated diseases. This interaction may play a major role in determining tumorigenicity and in forming clinical features of the disease. Increased provirus load found in patients with HAM/TSP and HU results from clonal expansion of the HTLV-1-infected T-cells, which has been implicated in the pathogenesis of HTLV-1-associated diseases. Regulatory mechanisms of clonal growth remain unknown. Efforts to characterize functions of the viral proteins and the virus-infected cells and to understand the natural history of the HTLV-1-infection are required to determine the mechanisms of HTLV 1 viral pathogenesis. PMID- 9401273 TI - Infections in patients with hematological diseases: recent advances in serological diagnosis and empiric therapy. AB - Recent advances in the diagnosis and therapy of infections in patients with hematological diseases are reviewed. In general, 40-60% of febrile episodes lack clinical or microbiological evidence of infection and are thus treated empirically. Among the cases of microbiologically documented bacteremia treated in our department, the incidence of Gram-negative bacteria was high (47.1%) and the incidence of Gram-positive bacteremia is increasing. To improve the diagnostic rate of Gram-negative sepsis, the measurement of plasma endotoxin was performed. Of 147 febrile neutropenic episodes, endotoxemia was observed in 58 (39.5%) and the causative microorganisms of these infections were deemed Gram negative bacteria. The measurement of plasma (1-->3)-beta-D-glucan, a ubiquitous component of fungi, was also performed for making early diagnosis of deep mycosis; the sensitivity of this assay was 90% and the specificity was 100%. The detection of (1-->3)-beta-D-glucan appears to be useful as a screening test of deep mycosis. The effects of the concomitant use of granulocyte-colony stimulating factor (G-CSF) and empiric antibiotic therapy for febrile neutropenia were studied in a randomized fashion. G-CSF did not affect the rate of the response to the empiric antibiotic therapy, although a significant effect of G CSF on neutrophil recovery was observed. Guidelines for empiric antibiotic and antifungal therapy combined with serological diagnosis are proposed. PMID- 9401274 TI - HLA DR2: a predictive marker in response to cyclosporine therapy in aplastic anemia. AB - The effect of immunosuppressive agents on HLA DR2-aplastic anemia (AA) has recently been investigated by different groups. In the present report, we analyzed 40 Turkish AA patients, who received immunosuppressive therapy (IST) and 12 AA's who were transplanted from HLA matched siblings. HLA DR2 frequency was 0.442 and significantly higher in AA's when compared to an unrelated healthy control group (RR: 2.93, 95% confidence interval 1.48-5.77, P = 0.001. Patients received antithymocyte or antilymphocytic globulin (AT/LG) or AT/LG plus cyclosporine-A (CsA) or CsA alone. In DR2+ and DR2- patients overall response rates were 73.3 and 30%, respectively (P = 0.03). When patients were analyzed separately, CsA administration either alone or in combination with AT/LG gave favorable results in the DR2+ group (P = 0.02). In contrast AT/LG presence alone was shown to be inadequate. PMID- 9401275 TI - Granulocyte colony-stimulating factor-combined marrow-ablative chemotherapy and autologous blood cell transplantation for the treatment of patients with acute myelogenous leukemia in first remission. The Fukouka Bone Marrow Transplant Group. AB - We conducted a clinical trial to increase the chemosensitivity of residual leukemic cells by combining G-CSF to marrow-ablative chemotherapy, including cytosine arabinoside (Ara-C), and facilitated by autologous blood cell transplantation (ABCT) for treatment of acute myelogenous leukemia (AML) in first complete remission. A total of 16 patients were consecutively treated with granulocyte colony-stimulating factor (G-CSF)-combined high-dose chemotherapy (busulfan, etoposide and Ara-C) followed by autotransplantation of peripheral blood progenitor cells, which had been collected after the consolidation chemotherapy. At a median follow-up time of 44.5 months, the probability of 5 year event-free survival was 74.5% with only three leukemic relapses. This preliminary observation suggests the effectiveness of G-CSF-combined conditioning and ABCT as a post-remission therapy for AML. PMID- 9401276 TI - Determining the optimal time for peripheral blood stem cell harvest by detecting immature cells (immature leukocytes and immature reticulocytes) using two newly developed automatic cell analyzers. AB - To determine the optimal time of peripheral blood stem cell harvest (PBSCH), we evaluated the usefulness of the detection of immature hematopoietic cells (immature leukocytes and reticulocytes) using newly developed automatic cell analyzers, and compared the results with those of conventional CD34 measurement by flow cytometry (FCM). The presence of immature leukocytes and reticulocytes was calculated as immature leukocyte information (IMI)-positive rate and high fluorescence reticulocyte (HFR)-positive rate by multi-item autoanalyzers. Both the IMI-positive rate and HFR-positive rate positively correlated with the CD34 positive rate and was more highly correlated with the colony forming units-mixed (CFU-Mix) count in peripheral blood. In addition, they positively correlated not only with the CD34-positive rate, but also with the colony forming units granulocyte/macrophage (CFU-GM) count and CFU-Mix count in harvested stem cell juice. These findings imply that the parameters of IMI and HFR could be used for determining the optimal time of PBSCH, and could also be useful for quantifying stem cells. PMID- 9401277 TI - Flow cytometric analysis of Thy-1 expression in CD34-positive acute leukemia. AB - We analyzed the expression of the Thy-1 antigen (CD90) in CD34+ acute leukemia using two-color flow cytometry. Leukemic cells were obtained from the bone marrow (BM) and/or the peripheral blood (PB) of 57 patients: 37 with acute myelogenous leukemia (AML) including nine with secondary AML following myelodysplastic syndrome (MDS/AML), and 20 with acute lymphoblastic leukemia (ALL) including three with chronic myelogenous leukemia in blast crisis (CML-BC) of the lymphoid type. Among these patients, one (3.6%) with de novo AML, two (22.2%) with MDS/AML, three (17.6%) with de novo ALL, and two (66.7%) with CML-BC coexpressed CD34 and Thy-1 (CD34+ Thy-1+) on more than 20% of the mononuclear cells within 'lymph' plus 'blast' window. Thy-1 was rarely expressed in de novo acute leukemia especially in AML. Interestingly, in 1 patient with CML-BC, the leukemic cells in BM were divided into two subpopulations (CD34+ Thy-1low and CD34+ Thy-1high), whereas most of the CD34+ leukemic cells in PB were Thy-1high. However, the mechanism for the mobilization of CD34+ Thy-1high leukemic cells into the PB is unknown. PMID- 9401278 TI - Rationale and hematologic target points in response-oriented individualized induction chemotherapy for acute myeloid leukemia. AB - Response-oriented individualized induction chemotherapy (ROIIC) has been reported to achieve a high complete remission (CR) rate in acute myeloid leukemia (AML). In ROIIC, drug dose is modified for each patient to achieve hematologic target points, based on the concept that this achievement, not a preset drug dose, increases the likelihood of attaining CR. However, the existing target points lack objectivity, therefore prohibiting worldwide use of ROIIC. In addition, the achievement of these target points does not statistically relate to CR, which contradicts the concept of ROIIC. To solve these problems, we evaluated various parameters of 117 AML cases treated by ROIIC to identify the factors relating to CR. Of 117 AML subjects, 75 achieved CR with ROIIC and most cases achieving CR did so with one ROIIC course. Not the administered dose of drugs but several hematologic parameters at the end of ROIIC correlated significantly with CR. A total of four parameters, all related to the leukemic cell mass, were independent predictive factors of CR. A formula consisting of these parameters predicted CR cases with high sensitivity (98%) and non-CR cases with high specificity (96%). Based on this formula, an objective target point (a bone marrow blast mass ratio between before and after ROIIC of below 0.15), valid for dose modification during ROIIC, was established. These results validate the concept of ROIIC and enable objective dose modification in ROIIC and universal application of this therapy. Further, the formula predicting non-CR cases at the end of ROIIC is useful for deciding early therapeutic intervention for such cases. PMID- 9401279 TI - Differentiation induction in non-lymphocytic leukemia cells upon treatment with mizoribine. AB - Inosine-5'-monophosphate (IMP) dehydrogenase catalyzes the rate-limiting reaction of guanine nucleotide biosynthesis and has been implicated in the reaction of cell growth and differentiation. We examined the ability of mizoribine, an IMP dehydrogenase inhibitor, to induce differentiation in HL-60 and U937 cells as well as in fresh leukemic blast cells from patients with non-lymphocytic leukemia. Treatment with mizoribine reduced intracellular GTP levels and induced morphologic and functional differentiation in these two cell lines in a dose dependent manner. HL-60 and U937 cells developed polymorphic nuclei and macrophage-like cytoplasm, respectively, as well as expression of CD11b and CD14 antigens and the ability to oxidize NBT. These changes became evident when intracellular GTP levels decreased to approximately 30% of untreated controls and were abrogated by addition of guanosine to the media. However, in fresh leukemic cells, the cells showing maturation in response to mizoribine were limited in those derived from two of ten patients having non-lymphocytic leukemia. These findings suggest mizoribine could induce differentiation in HL-60 and U937 cells through a decrease of intracellular GTP levels. Further study is required to determine its clinical use in patients with acute non-lymphocytic leukemia. PMID- 9401280 TI - Correlation between immunological abnormalities and prognosis in myelodysplastic syndrome patients. AB - Immunological abnormalities (IA) are frequently observed in patients with myelodysplastic syndromes (MDS). Although there have been a number of analyses of the prognostic factors, there have been few studies, if any, to determine whether IA affects prognosis. We investigated the prognosis of 153 MDS patients with or without IA who were treated at a single Japanese institute for 10 years. Nineteen of 153 patients (12%) developed autoimmune disorders. One hundred of 153 patients (63%) had an abnormality in at least one immunological laboratory test. Hypergammaglobulinemia was found in 50 of 128 (39%) patients tested, hypogammaglobulinemia was observed in 10 of 128 (8%), positivities of antinuclear antibody, RA factor. DNA antibody, and direct antiglobulin test were observed in 30%, 14%, 7% and 12%, respectively, and a CD4/CD8 ratio < 1 was observed in 20%. There was no significance in the distribution of age, MDS subtype, or sex between patients with and without IA. The survival of MDS patients without IA was significantly better than that of patients with IA, and the survival of patients with a CD4/CD8 ratio > 1 was also significantly superior to the survival of those with an inverted CD4/CD8 ratio. Patients with IA tended to die of infection or leukemic progression in comparison with those without IA, suggesting that IA may be associated with susceptibility to opportunistic infection and disease progression. PMID- 9401281 TI - Hypercalcemia in children presenting with acute lymphoblastic leukemia. AB - Although hypercalcemia is a well-recognized complication in malignant disorders, neither the incidence and prognostic significance of hypercalcemia, nor the role of parathormone related peptide (PTHrP) in pediatric acute lymphoblastic leukemia (ALL) have been clarified. Of 83 newly diagnosed pediatric ALL patients with early pre-B cell phenotype treated at our hospital during the last 8 years, four patients were diagnosed as having hypercalcemia (> 14 mg/dl). In these 4 hypercalcemic ALL patients at onset, serum calcium levels ranged from 14.6 to 20.8 mg/dl (normal 7.4-9.0 mg/dl), and serum PTHrP levels were markedly elevated to 112-240 pmol/l (normal range: 17.6-61.2 pmol/l). Unlike patients with ordinary ALL in childhood, gastrointestinal symptoms (nausea, vomiting, abdominal pain) and skeletal symptoms (bone pain, gait disturbance) were the chief complaints. Because of these characteristic symptoms, bone marrow aspiration was carried out in two patients in an attempt to diagnose ALL before leukemic cells appeared in peripheral blood. Serum calcium levels were promptly normalized by induction chemotherapy. The four patients have been in complete remission from 35+ to 125+ months. Based on these results, the incidence of hypercalcemia in pediatric ALL patients with early pre-B cell phenotype at our institute is calculated to be about 4.8%. Gastrointestinal and skeletal problems are the characteristic initial symptoms, and hypercalcemia does not seem to be significant in the prognosis of these patients. PMID- 9401282 TI - A case of mu-heavy chain disease: combined features of mu-chain disease and macroglobulinemia. AB - A case of mu-heavy chain disease (HCD) is described. The patient, a 40-year-old man, presented with an intracranial tumor. The bone marrow of this patient showed infiltration with both plasma cells and lymphocytes. The majority of plasma cells were vacuolated and the result of immunoelectrophoresis of serum protein revealed an arc with anti-IgM antiserum and an additional rapid migrating arc of different mobility with anti-kappa antiserum. The urine contained a kappa-type Bence Jones protein. An enzyme-linked antibody study showed that the majority of plasma cells in bone marrow contained both mu and kappa antigenic determinants in their cytoplasm. On Sephadex G-200 gel filtration, the monoclonal IgM-kappa protein and the mu-chain fragment were detected in the serum, suggesting the combined features of mu-HCD and macroglobulinemia. The molecular weight of the mu-chain fragment was approximately 45,000 daltons. The intracranial tumor completely disappeared after irradiation therapy. However, he died 1 year later after development of a huge abdominal tumor. PMID- 9401283 TI - Serum soluble IL-6 receptor concentrations correlate with stages of multiple myeloma defined by serum beta 2-microglobulin and C-reactive protein. AB - Serum soluble interleukin-6 receptor (sIL-6R) concentrations were measured in 52 patients with multiple myeloma (MM) and 24 normal controls, using a commercially available immunoenzymatic assay kit. Patients were staged according to the Bataille et al. myeloma staging system based on the levels of patients' serum beta 2-microglobulin and C-reactive protein. Twenty-one patients were at stage A of disease, 19 at stage B and 12 at stage C at the time of serum collection for sIL-6R determination. Serum sIL-6R concentrations ranged from 15 to 176 ng/ml with a mean of 64.8 +/- 35.9 ng/ml and a median of 58 ng/ml in the entire group of patients studied. These values were significantly higher than those of 34.4 +/ 13.4 ng/ml found in the controls (P < or = 0.001). Patients of stage C had higher sIL-6R levels (94.8 + 41.2 ng/ml) than patients of stage B (67.7 +/- 31.0 ng/ml) (P < 0.01), and markedly higher than patients of stage A (45.0 +/- 23.1 ng/ml) (P < 0.001). Serum levels of sIL-6R in patients with stage A disease did not differ statistically from those of the controls. A linear positive correlation was observed between serum levels of the receptor and the stage of MM (r = 0.539, P < 0.001). These data strongly suggest that serum sIL-6R concentrations correlate with the stages of MM and may be used as an indicator of the activity of the disease. PMID- 9401284 TI - Effect of thrombopoietin (c-Mpl ligand) alone and in combination with other hematopoietic growth factors on human megakaryocytopoiesis in serum-free cultures. AB - The effect of human recombinant (hr) thrombopoietin (TPO) on human megakaryocytopoiesis was studied in a serum-free system. hrTPO induced megakaryocyte colony formation by purified CD 34-positive cells and polyploidization of megakaryocytes by purified CD41a-positive cells. hrTPO gave rise to much smaller colonies which appeared at an earlier time compared to the use of human recombinant interleukin-3 (hrIL-3), suggesting that hrTPO predominantly affects the population of megakaryocyte progenitor cells in the late stage. hrIL-3 additively increased the hrTPO-induced megakaryocyte colony formation by CD34-positive cells. The hrTPO-induced megkaryocyte colony formation was also increased by the presence of hrIL-6, hrIL-11, human recombinant erythropoietin (hrEpo) or human recombinant stem cell factor (hrSCF), none of which stimulated megakaryocyte colony growth when added alone. The combined addition of hrTPO, hrIL-3 and hrSCF to CD34-positive cells markedly stimulated megakaryocyte colony formation and produced large numbers of megakaryocytes. hrTPO stimulated the polyploidization of CD34-positive cell-derived megakaryocytes in liquid culture. However, the addition of hrIL-6, hrIL-11 or hrEpo to hrTPO did not further enhance the hrTPO-induced polyploidization. These findings indicate that at the megakaryocyte progenitor cell level, the effect of hrTPO can be promoted by the presence of various hematopoietic growth factors involved in human megakaryocytopoiesis. PMID- 9401285 TI - Rapid progression of chronic myelomonocytic leukemia following diaminodiphenyl sulphone treatment for dermatitis herpetiformis. AB - A 41-year-old patient with dermatitis herpetiformis (DH) developed steroid resistant blebs as a sign of exacerbating DH. The skin symptoms were resolved after 2 weeks of oral administration of diaminodiphenyl sulphone (DDS). However, 3 weeks after the start of DDS, he suffered from edematous eruption on the cheeks and neck, enlargement of the pharynx, systemic lymphoadenopathy and hepatomegaly. In addition, his leukocyte count increased rapidly from 10.1 x 10(9)/l with 13% monocytes just before the start of DDS, to 24.6 x 10(9)/l with 28% monocytes. Bone marrow aspirate showed trilineage dysplasia and chronic myelomonocytic leukemia (CMML) was diagnosed. The patient died from septic shock during neutropenia following cytotoxic chemotherapy. In this case, CMML was complicated with DH and the administration of DDS accelerated the progression of CMML with the manifestations of DDS syndrome. Although DDS is a well-established drug for DH, DDS should be used with great caution when a hematological malignancy coexists. PMID- 9401286 TI - Distribution of endogenous endonuclease activities capable of producing internucleosomal DNA cleavage in hematopoietic cells. PMID- 9401287 TI - Kaposi's sarcoma--associated herpesvirus-like DNA sequences are not present in adult T-cell leukemia. PMID- 9401288 TI - Glutathione S-transferase theta 1 gene (GSTT1) defect in Japanese patients with myelodysplastic syndromes. PMID- 9401290 TI - Pelton & Crane Light alert. PMID- 9401289 TI - Poor kids' teeth: a crying shame. PMID- 9401291 TI - Dental benefits issues. PMID- 9401292 TI - Dental hygiene. PMID- 9401293 TI - CDA opposes proposed HBV recommendations. PMID- 9401294 TI - Health Canada issues warning letter on lead shielding. PMID- 9401295 TI - Supreme Court of Delaware rules in favor of HIV-positive dentist. PMID- 9401296 TI - The business of dentistry. PMID- 9401297 TI - Guidelines on the use of space maintainers following premature loss of primary teeth. AB - OBJECTIVE: To formulate evidence-based guidelines on the appropriate use of space maintaining appliances to prevent or reduce the severity of malocclusion in the permanent dentition following the premature loss of primary teeth. OPTIONS: Placement of lingual arch, palatal arch, band-loop, crown-loop, and intra alveolar space maintainers. OUTCOMES: Reduced prevalence or severity of space loss in the primary or mixed dentition, reduced prevalence or severity of malocclusion in the permanent dentition measured as significant changes in: crowding, ectopic eruption, impacted teeth, Angle's class II or III occlusion, crossbite, deep overbite, deep overjet, or midline shift. EVIDENCE: Articles published from 1966 to 1996, located though Medline searches. Only clinical trials, cohort studies, case-control studies, or large case series were considered. VALUES: Relevant clinical findings were evaluated and categorized using evidence-based methods and values established by the Canadian Task Force on the Periodic Health Examination. A recommendation was developed for the use of space maintainers. BENEFITS, HARMS AND COSTS: The potential benefits of using space maintaining appliances include reduced prevalence or severity of: crowding, ectopic eruption, tooth impaction, crossbite, excessive overbite and overjet, and poor molar relationship. Other advantages include the potential for considerable cost savings by reducing the need for future orthodontic treatment. The potential disadvantages of using space maintaining appliances include soft tissue impingement, interference with eruption of adjacent teeth, pain, plaque accumulation, caries, and broken, dislodged or lost appliances. RECOMMENDATIONS: There is poor evidence to recommend for or against the use of space maintainers to prevent or reduce the severity of malocclusion in the permanent dentition (see Table 1, Recommendation C) Decisions regarding the use of space maintainers must therefore be guided by factors other than scientific evidence. PMID- 9401298 TI - Anatomically correct soft tissue profiles using fixed detachable provisional implant restorations. AB - For the past decade, the dental profession has given very little thought to the design of fixed detachable restorations with anatomically correct emergence profiles. Over-contoured implant restorations are frequently seen, and they may be challenging for patients to maintain. This article will address the most common techniques used to improve the soft tissue profile, and will show how implant-borne provisional restorations allow ideal soft tissue profiles to be achieved. PMID- 9401299 TI - Denturists: do they really provide more affordable care in Ontario? AB - The 1996 Denturist Association of Ontario fee guide and the Ontario Dental Association Fee Guide for General Practitioners were examined to identify variations in the fees charged for a range of removable prosthodontic services. Fee guides were selected for this analysis as third-party insurers and government dental plans frequently use them to establish fee schedules and levels of reimbursement. However, it is recognized that dentists set their own fees, which may be higher or lower than the fees suggested in the guide. Although the descriptions of the specific services listed in the guides were similar in many cases, no attempt was made to examine variations in the quality of care provided by dentists and denturists, or the approach to treatment offered by their respective professions. The analysis revealed that a number of procedure fees were, on average, 15 per cent higher in the Ontario Dental Association (ODA) fee guide compared to the denturists' fee guide. However, a wide range of prosthetic services, including partial dentures, were less expensive in the ODA fee guide. Based on this analysis, there appears to be no substantial cost differential between the services provided by dentists and denturists six years after the proclamation of the new Regulated Health Professions Act in Ontario, as the denturists have claimed. The denturist association's further claims of greater choice and improved access may also be questionable, and should be reexamined in light of these findings. PMID- 9401300 TI - Dental unit waterline microbiology: a cautionary tale. AB - The question of dental unit waterline contamination now concerns the dental profession on a number of levels and has become part of the landscape in dentistry. Researchers have identified the problem, studied it, and published their results. Official organizations have reacted to the problem by issuing press releases and recommendations. In the mean time, dental companies have jumped into the fray, ensuring a proliferation of products designed to mitigate a problem that certain people feel is imaginary are artificially inflated. Despite the number of publications describing waterline contamination, we still face the challenge of determining whether are picture of the problem is truly accurate. For example, it has become almost customary to use the term contamination when talking about waterlines, even though it is inadequate. In addition, we are moving ahead with water quality standards for dental units at a time when certain fundamental questions remain unanswered. What should we be measuring and what methods should we be using? Do certain disinfection procedures have an opposite effect to the one desired? Finally, the question of health risks linked to the colonization of waterlines has not been adequately addressed by researchers. PMID- 9401301 TI - Stabilization of rubber dam for child patients and fibre-reinforced postorthodontic retainer. PMID- 9401302 TI - Getting what we are worth. PMID- 9401304 TI - Nursing centers--models of professional practice. PMID- 9401303 TI - Future organizational leadership. PMID- 9401305 TI - Fighting success--facilitation of senior faculty. PMID- 9401306 TI - Education and practice partnerships in California. AB - As the most populous state in the United States, California often serves as a bellwether of events to come. The following describes nursing work force issues in California, examples of how nurse educators are responding to health care system changes, and how practice colleagues are joining educators to prepare nurses for meeting the health needs of the people of California. The discussion begins with historical perspectives from the late 1980s to the present, the founding of the California Strategic Planning Committee for Nursing and its work, the Robert Wood Johnson Colleagues in Caring project for developing a master plan for nursing in California, and examples of existing programs in California that serve as models for preparing the best care providers to meet the health care needs of the population. PMID- 9401307 TI - Student and preceptor perceptions of factors in a successful learning partnership. AB - Thirty-two registered nurse preceptors and 42 senior undergraduate nursing students completed a survey ranking factors related to both participants in the clinical learning partnership. Mann-Whitney U-Wilcoxon Rank Sum W tests showed statistically significant differences in the ranking of four factors (the ability to give and receive criticism, knowledge of the preceptoring process, clinical competence, and compatibility) that contribute to successful learning partnerships. Nurse educators and professional nurses should acknowledge these perceptual differences and include these in student and preceptor orientation programs to promote a positive teaching and learning partnership. PMID- 9401308 TI - Mid-life women doctoral students rediscover "voice" in a community of scholarly caring. AB - Recent research indicates that doctoral study is for many "an excessively painful rite of passage" for which an "indefensible human price is exacted." For mid-life women doctoral students in nursing and education doctoral programs, this human price is sometimes paid in the currency of loss of personal voice. Because they often assume leadership positions after graduation, it is a significant loss to the professions when these women's mature voices are lost or even temporarily silenced. This article describes the design, findings, evaluation, and implications of an emancipatory inquiry entitled "Gifted Women: Doctoral Study As Heroic Journey." Simultaneously a participatory research project and curricular innovation, this unique 18-month project joined facilitators and participants as coresearchers in a workshop and four reunions that integrated feminist process with expressive, esthetic approaches. The results of this emancipatory inquiry showed that viewing doctoral study as a heroic journey enlightened these mid-life women participants to reclaim their personal voices, empowered them with the support of a community of scholarly caring, and emancipated them to undertake personally meaningful and scholarly dissertation research. PMID- 9401309 TI - Cognitive pathways and historical research. AB - The nursing literature is replete with articles detailing the logical reasoning processes required by the individual scientist to implement the rigors of research and theory development. Much less attention has been focused on creative and critical thinking as modes for deriving explanations, inferences, and conclusions essential to science as a product. Historical research, as a particular kind of qualitative research, is dependent on and compatible with such mental strategies as logical, creative, and critical thinking. These strategies depict an intellectual framework for the scientist examining archival data and offer a structure for such inquiry. A model for analyzing historical data delineating the cognitive pathways of logical reasoning, creative processing, and critical thinking is proposed. PMID- 9401310 TI - Management of comorbid depression and heart disease: the time has (almost) come. PMID- 9401312 TI - Post-traumatic stress disorder and serotonin: new directions for research and treatment. AB - The overlap in clinical phenomenology and morbidity between post-traumatic stress disorder (PTSD) and such conditions as major depression, anxiety disorders and aggression, in which a serotonin dysfunction is implicated, suggests a role for serotonin in the pathophysiology of PTSD. In this paper, we review current knowledge concerning the role of serotonergic mechanisms and interventions in PTSD. Since there is no clearly effective pharmacologic intervention for this disorder, the underlying neurochemical dysfunction needs to be carefully defined so that more effective treatment can be developed. Preclinical and clinical studies of the serotonergic mechanisms in the pathophysiology of PTSD and treatment trials involving serotonergic agents are limited, but indicate considerable promise. Further investigation of a serotonergic dysfunction in PTSD and of its treatment with serotonergic agents is warranted. PMID- 9401313 TI - Refractory depression: treatment strategies, with particular reference to the thyroid axis. AB - In the last few years, it has become evident that major depressive disorder often runs a chronic and recurrent course. Early effective intervention may increase the liklihood of a good long-term prognosis. The main treatment options for patients who fail to respond to antidepressant therapy and the relative advantages of each are critically reviewed. These include substitution, replacing one antidepressant with another, and augmentation/combination, in which a second antidepressant is added to the first. Particular emphasis is placed on the role of triiodothyronine (T3) in augmentation therapy. The theoretic rationale for using augmentation/combination therapy and its relative advantages and disadvantages over substitution therapy are critically reviewed. PMID- 9401314 TI - Effects of citalopram treatment on behavioural, cardiovascular and neuroendocrine response to cholecystokinin tetrapeptide challenge in patients with panic disorder. AB - Eight patients with panic disorder were administered 20 micrograms of cholecystokinin tetrapeptide (CCK-4) before and after 8 weeks of treatment with the selective serotonin reuptake inhibitor (SSRI) citalopram. All patients responded to treatment by showing a significant general improvement and reaching a panic-free state for 2 weeks. At the rechallenge with CCK-4, patients displayed a marked reduction in the intensity and number of panic symptoms. The frequency of panic attacks induced with CCK-4 decreased by 50% after treatment. Citalopram treatment had no substantial effect on cardiovascular (heart rate and blood pressure) or hormonal (cortisol, prolactin and growth hormone) responses to CCK 4. Patients who still had panic attacks after treatment demonstrated a blunted growth hormone response to CCK-4 that was not seen in those who did not have panic attacks. This study suggests that treatment with an SSRI can reduce an enhanced sensitivity to CCK-4 without modifying cardiovascular and neuroendocrine responses to CCK-4 in patients with panic disorder. PMID- 9401316 TI - Successful treatment of steroid-induced panic disorder with fluvoxamine. PMID- 9401315 TI - Effective treatment of mania with olanzapine: 2 case reports. PMID- 9401311 TI - Galanin-acetylcholine interactions in rodent memory tasks and Alzheimer's disease. AB - Galanin is a 29-amino-acid neuropeptide that is widely distributed in the mammalian central nervous system. Galanin-immunoreactive cell bodies, fibres and terminals, and galanin binding sites, are located in the basal forebrain of rats, monkeys and humans. Galanin fibres hyperinnervate the surviving cholinergic cell bodies in patients with Alzheimer's disease (AD). In rats, galanin inhibits acetylcholine release and produces deficits in learning and memory. These findings suggest that overexpressed galanin may contribute to the cognitive impairments exhibited by patients with AD. This paper reviews the literature on galanin distribution and function in light of its putative role in the mnemonic deficits in patients with AD, the effects of galanin on tests of learning and memory, and preliminary experiments with galanin antagonists in animal models of AD. PMID- 9401317 TI - Distinguishing anxiety disorders psychometrically. PMID- 9401318 TI - Mania in children with PDD. PMID- 9401319 TI - ADHD/thyroid dysfunction. PMID- 9401320 TI - Immunology of TS/OCD. PMID- 9401321 TI - Pemoline in ADHD. PMID- 9401322 TI - Childhood mania in India. PMID- 9401323 TI - Domestic violence and psychopathology. PMID- 9401324 TI - Behavioral effects of hypoglycemia. PMID- 9401325 TI - Looking at the future through orange-colored glasses. PMID- 9401326 TI - Mental retardation: a review of the past 10 years. Part I. AB - OBJECTIVE: To review the literature over the past decade on mental retardation, particularly as regards its definition, prevalence, major causes, and associated mental disorders. METHOD: A computerized search was performed for articles published in the past decade, and selected papers were highlighted. RESULTS: The study of mental retardation has benefited considerably by advances in medicine generally and by developments in molecular neurobiology in particular. Increasing awareness of psychiatric comorbidity in the context of intellectual disability highlights the need for studies of the phenomenology and treatment of mental disorders in this population. CONCLUSIONS: Although the study of developmental disorders has advanced significantly over the past decade, considerable work remains. Mental retardation is a model for the utility of the biopsychosocial approach in medicine. PMID- 9401327 TI - Mental retardation: a review of the past 10 years. Part II. AB - OBJECTIVE: To review the literature over the past decade on mental retardation, particularly with respect to genetics and behavioral phenotypes. METHOD: A computerized search was performed for articles published in the past decade, and selected papers were highlighted. RESULTS: The study of mental retardation has benefited considerably by advances in medicine generally, and by developments in molecular neurobiology in particular. These advances in genetics have led to new insights regarding the causes of mental retardation, as well as a growing appreciation of behavioral phenotypes associated with some mental retardation syndromes. CONCLUSIONS: Although the study of developmental disorders has advanced significantly over the past decade, considerable work remains. Mental retardation should remain the model for the utility of the biopsychosocial approach in medicine. PMID- 9401328 TI - Evolution and revolution in child psychiatry: ADHD as a disorder of adaptation. AB - Current knowledge about early plasticity and children's responsiveness to environmental modifications as well as the atheoretical nature of current nosological systems necessitate alternative models to explain the phenomena of childhood behavioral and emotional disturbances. Evolutionary biology provides one such framework. It organizes data from the behavioral and cognitive sciences and parallels similar efforts in other areas of medicine and biology. Through an evolutionary biological lens, some mental disorders are better viewed as an adaptive response to early pathogenic environments and/or reflect the optimization of brain function to some environments at the cost of poorer response to the demands of other environments. As an example, the authors examine attention-deficit/hyperactivity disorder (ADHD) in relation to evolutionary theories of psychology and biology and clarify the potentially adaptive nature of characteristics of inattention, impulsivity, and motoric hyperactivity, depending on the nature of child's environments. Reframing ADHD characteristics according to evolutionary theory has important treatment implications for clinicians and offers researchers opportunities for novel scientific discoveries. PMID- 9401329 TI - Correspondence between DSM-III-R and DSM-IV attention-deficit/hyperactivity disorder. AB - OBJECTIVE: To evaluate the correspondence between DSM-III-R and DSM-IV definitions of attention-deficit/hyperactivity disorder (ADHD) in clinically referred children. Results of the field trials led to the hypothesis that there would be a strong correspondence between DSM-III-R and DSM-IV subtypes. METHOD: The sample consisted of all children and adolescents consecutively referred to a pediatric psychopharmacology clinic (N = 405). Children were comprehensively evaluated with structured diagnostic interviews assessing both DSM-III-R and DSM IV ADHD. DSM-III-R symptoms were used to approximate DSM-IV subtypes. Kappa statistics and conditional probabilities were used to examine the correspondence between DSM-III-R and DSM-IV ADHD. RESULTS: Ninety-three percent of children who received a DSM-III-R diagnosis of ADHD also received a DSM-IV ADHD diagnosis. The kappa coefficient assessing the agreement between DSM-III-R and DSM-IV ADHD was .73 (z = 14.6, p < .0001). The kappa coefficient assessing the agreement between the DSM-III-R-approximated subtypes and the actual DSM-IV subtypes was .71 (z = 15, p < .0001). CONCLUSION: These results confirm previous findings and indicate that the change from DSM-III-R to DSM-IV results in minimal changes in case identification and provides support for diagnostic continuity between the two classification systems. PMID- 9401330 TI - Noradrenergic mechanisms in ADHD children with and without reading disabilities: a replication and extension. AB - OBJECTIVE: To examine noradrenergic (NA) function in children with attention deficit hyperactivity disorder (ADHD) by replicating and expanding upon a previous finding that ADHD children with and without reading disabilities (RD) differ in plasma levels of the NA metabolite 3-methoxy-4-hydroxyphenylglycol (MHPG). METHOD: Plasma levels of MHPG were compared in ADHD children who were subdivided on the basis of the presence or absence of RD. Subsequently, this replication sample was combined with a previously studied sample to further explore the relationship between plasma MHPG levels and measures of cognitive function in children with ADHD. RESULTS: Plasma levels of MHPG were significantly lower in ADHD children without RD, compared with those with RD, replicating a published finding. Analyses in the combined sample indicated that, among children with ADHD, plasma MHPG levels were inversely associated with measures of academic achievement and verbal processing, but not parent or teacher ratings of behavior or continuous performance test measures of attention and impulsivity. CONCLUSIONS: These data indicate that children with ADHD are not homogeneous with regard to NA function and that neurochemical variation is closely associated with differences in clinical characteristics of the children. PMID- 9401331 TI - Affective valence and arousal in ADHD and normal boys during a startle habituation experiment. AB - OBJECTIVE: To measure two dimensions of emotion (affective valence and arousal) in 29 boys with attention-deficit hyper-activity disorder (ADHD) and 32 normal boys. METHOD: After a startle habituation experiment during which these subjects heard 40 startling sounds while watching a silent interesting movie, they were asked 12 questions (categorized a priori into questions relating to affective valence and to arousal) about their emotional reactions to these putatively unpleasant and pleasurable stimuli. Responses were recorded for the two dimensions of emotion, using two cartoon strips in each of which five expressions of a cartoon character varied linearly from happy to unhappy (affective valence dimension) and calm to excited (arousal dimension). RESULTS: Factor analyses of the 12 responses revealed four factors in which the highest loadings were for affective valence to the startle responses, affective valence to the silent movie, arousal, and scary feelings. Relative to the normal group, the responses of the ADHD group were significantly biased toward pleasurable valence to the startling stimuli and to the silent movie, with a trend toward hypoarousal. Startle magnitude and habituation were similar in both groups. The normal tonic heart rate acceleration throughout the experimental session was not sustained in the ADHD group. CONCLUSIONS: The self-reports of affective valence biased in the direction of pleasure and away from displeasure and the trend toward hypoarousal suggest an emotional dysfunction in ADHD. PMID- 9401332 TI - Gender differences in children with ADHD, ODD, and co-occurring ADHD/ODD identified in a school population. AB - OBJECTIVE: To examine gender differences among children with disruptive behavior disorders (DBDs) from an ethnically diverse school sample. METHOD: From 2,984 children, children with attention-deficit/hyperactivity disorder, combined type (ADHD-C) (46 boys, 11 girls), oppositional defiant disorder (ODD) (59 boys, 35 girls), and co-occurring ADHD-C/ODD (76 boys, 27 girls), diagnosed by teacher rated DSM-IV symptoms, were compared with each other and with 254 controls on teacher ratings of symptoms, social functioning and Achenbach Teacher's Report Form scales. RESULTS: Children with ADHD-C/ODD received the poorest ratings on all variables. In "pure" groups, children with ODD were rated as learning more, working harder, and being less inattentive than children with ADHD-C; only the ODD group showed more internalizing problems than controls. For ADHD-C and ODD groups, ratings of aggression and some individual symptoms were higher in boys than girls. Girls with ODD were rated as more appropriate and less inattentive, but unhappier and more socially impaired than boys with ODD. Overall, girls received higher peer dislike scores than boys. CONCLUSIONS: Comorbidity and gender issues affect the correlates of DBDs, with learning problems higher in ADHD-C, internalizing problems associated only with ODD, and greatest impairment for ADHD-C/ODD groups. Despite having similar or less behavioral dysfunction, girls with DBDs may have more social problems than boys with DBDs. PMID- 9401333 TI - Impact of comorbid oppositional or conduct problems on attention-deficit hyperactivity disorder. AB - OBJECTIVE: To investigate whether the presence of comorbid oppositional defiant disorder (ODD) or conduct disorder (CD) alters the correlates of attention deficit hyperactivity disorder (ADHD). METHOD: Three groups of children (33 "pure" ADHD, 46 ADHD + ODD, and 12 ADHD + CD) were compared on measures of ADHD, aggression, anxiety, parental psychopathology, self-esteem, school, and social emotional functioning. RESULTS: Findings indicated that the presence of comorbid oppositional or conduct problems in children with ADHD altered the correlates of ADHD across a number of areas, including greater ADHD symptom severity and social dysfunction. Nevertheless, some correlates were more closely linked with the comorbid condition of ADHD + CD (e.g., higher aggression, anxiety, and maternal pathology, as well as decreased self-esteem), while others appeared more closely linked with ADHD + ODD (e.g., social withdrawal, elevated academic achievement paired with higher perceived scholastic competence). CONCLUSIONS: Findings support the distinctive profiles of the disruptive behavior disorder groups and emphasize the deleterious effects on the quality of life experienced by the comorbid conditions. The need for syndrome-specific interventions is stressed. PMID- 9401334 TI - A prospective study of hyperactive boys with conduct problems and normal boys: adolescent and adult criminality. AB - OBJECTIVE: To examine the relationship between attention deficit disorder with hyperactivity in childhood and criminality in adolescence and adulthood in 89 hyperactive and 87 normal control subjects. METHOD: In this prospective study, adolescent follow-up intervals ranged from 13 to 21 years and adult follow-up ranged from 18 to 23 years. The official arrest records for all subjects were obtained. RESULTS: Hyperactive subjects had significantly higher juvenile (46% versus 11%) and adult (21% versus 1%) arrest rates. Juvenile and adult incarceration rates were also significantly higher. Childhood conduct problems predicted later criminality, and serious antisocial behavior in adolescence predicted adult criminality. CONCLUSIONS: Hyperactive children are at risk for both juvenile and adult criminality. The risk for becoming an adult offender is associated with conduct problems in childhood and serious antisocial behavior (repeat offending) in adolescence. Hyperactive children who do not have conduct problems are not at increased risk for later criminality. PMID- 9401335 TI - Childhood cancer: a two-year prospective study of the psychological adjustment of children and parents. AB - OBJECTIVE: To follow prospectively the psychological adjustment of young children, parents, and families during the first 2 years after the children's diagnosis of cancer. METHOD: Children aged 2 to 5 years with cancer diagnoses and their parents and families (n = 38) were assessed immediately after diagnosis, 1 year after diagnosis, and 2 years after diagnosis. At each assessment, the psychological adjustment of the children and their families was compared with the adjustment of a cohort of children and families in the general community (n = 39). RESULTS: Children with cancer and their parents experienced significantly more emotional distress than children and parents in the community during the period immediately after diagnosis. However, the number of problems experienced by the children with cancer and their parents declined during the first year after the children's diagnosis and stabilized at a level comparable with that found among children and parents in the general community. CONCLUSION: Although the results are consistent with reports that suggest that in the longer term the prevalence of psychological problems among children with cancer is similar to that found among children in the general community, they also highlight the considerable distress experienced by children and parents during the period immediately after the children's diagnosis. PMID- 9401336 TI - Traumas and other adverse life events in adolescents with alcohol abuse and dependence. AB - OBJECTIVE: Clinical observation suggests that adolescents with alcohol use disorders often have complex histories that include childhood maltreatment and other traumas. The aim of this study was to determine the relationships among adolescent alcohol use disorders and a broad range of traumas and adverse life events. METHOD: The subjects were 132 adolescents with alcohol dependence, 51 adolescents with alcohol abuse, and 73 adolescents recruited from the community as a control group. Trauma history was assessed by a semistructured interview and other adverse life events by questionnaire. RESULTS: Traumatic events reflecting interpersonal violence had occurred in many of the adolescents with alcohol dependence and abuse and few of the control adolescents. Adolescents with alcohol abuse or dependence, compared with control subjects, were 6 to 12 times more likely to have a physical abuse history and 18 to 21 times more likely to have a sexual abuse history. Sexual abuse was more common in females, and victimization by other violent acts was more common in males. Many other adverse life events were also significantly more common in the alcohol use disorder groups than in the control group, including having a close friend die, arguments within the family, and legal difficulties. CONCLUSIONS: These results demonstrate that trauma and other adverse life events are strongly associated with alcohol use disorders in adolescents. Clinical screening of adolescents with alcohol use disorders for a range of traumatic events is recommended. PMID- 9401337 TI - Axis II psychopathology as a function of Axis I disorders in childhood and adolescence. AB - OBJECTIVE: To examine the occurrence of elevated personality disorder (PD) dimensional scores in a community sample of young adults as a function of the occurrence of Axis I disorders through age 18 years. METHOD: 299 individuals who had been interviewed regarding Axis I disorders twice while in adolescence (first when 14 through 18 years of age) were carefully assessed regarding Axis I and II psychopathology at age 24. RESULTS: The prevalence of PD diagnoses was relatively low (3.8% in participants with a history of Axis I versus 1.7% in participants with no Axis I history). The occurrence of all four Axis I diagnostic categories (major depression, anxiety disorders, disruptive behavior disorders, substance use disorders) in childhood and adolescence was associated with elevated PD dimensional scores. The likelihood of elevated PD dimensional scores increased as a function of the number of Axis I disorders. Elevated PD scores were significantly associated with a negative course of major depression. CONCLUSIONS: Although the rates of PDs were low, the findings suggest a substantial degree of association between early-onset Axis I disorders and Axis II psychopathology in young adulthood. More research is needed to develop assessment and treatment recommendations addressing the early manifestations of PDs. PMID- 9401338 TI - Antecedents of preschool children's internalizing problems: a longitudinal study of low-income families. AB - OBJECTIVE: To examine antecedents of young children's internalizing problems using research related to emotion regulation to guide prediction. METHOD: Longitudinal data were collected on 86 low-income mother-child dyads to examine risk factors related to early internalizing problems as measured by the Child Behavior Checklist (CBCL). RESULTS: The following risk factors, assessed during infancy, were related to the development of preschool-age internalizing problems: negative emotionality, disorganized attachment classification, negative life events, exposure to child-rearing disagreements, and parenting hassles. In addition, the interaction of high negative emotionality and exposure to parental conflict added unique variance to the prediction of scores on the CBCL Withdrawal and Depression/Anxiety subscales. CONCLUSIONS: Children's preschool-age internalizing problems can be identified during infancy from multiple domains related to the development of emotion regulation. Further longitudinal work is encouraged that incorporates direct measurement of children's negative emotionality, parenting, and family factors that influence both parenting and children's emotion regulation. PMID- 9401339 TI - Is psychopathology associated with the timing of pubertal development? AB - OBJECTIVE: This investigation tested whether the timing of pubertal development was associated with concurrent and prior experiences of psychopathology (symptoms and disorders) in adolescent boys and girls. METHOD: A large (N = 1,709) community sample of high school students were interviewed using the Schedule for Affective Disorders and Schizophrenia for School-Age Children as adapted for use in epidemiological studies. Adolescents also completed a questionnaire battery covering a range of psychosocial variables. RESULTS: Analyses tested whether pubertal timing was associated with present and lifetime history of mental disorders, psychological symptoms, and psychosocial functioning. As hypothesized, early-maturing girls and late-maturing boys showed more evidence of psychopathology than other same-gender adolescents. CONCLUSIONS: Early-maturing girls had the poorest current and lifetime history of adjustment problems, indicating that this pattern of pubertal development merits attention by mental health providers and researchers. PMID- 9401340 TI - AACAP Official Action. Summary of the practice parameters for child custody evaluation. AB - This summary is presented as a guide for clinicians evaluating the often delicate and complex issues surrounding a child custody dispute. The historical basis of child custody and the various judicial presumptions that have guided courts, as well as the differences between performing child custody evaluation and engaging in traditional clinical practice, are reviewed in the complete document. Issues that are common to all child custody disputes are presented, including continuity and quality of attachments, preference, parental alienation, special needs of children, education, gender issues, sibling relationships, parents' physical and mental health, parents' work schedules, parents' finances, styles of parenting and discipline, conflict resolution, social support systems, cultural and ethnic issues, ethics and values, and religion. In addition, special issues that complicate custody evaluations are presented, including infants in custody disputes, homosexual parents, grandparents' rights, parental kidnapping, relocation problems, allegations of sexual abuse, and advances in reproductive technology, such as frozen embryos, oocyte donation, and artificial insemination. PMID- 9401341 TI - Back to the future: revisiting the 40-63-1988 syndrome. PMID- 9401342 TI - Communicating with our patients: the goal of bioethics. AB - Listening, teaching, understanding, exploring, explaining: these are the foundations of a sound patient-physician relationship. From these skills, we can then proceed to discussions on difficult topics such as preferences for end-of life care. We can share bad news without destroying hope. We can show what makes the medical profession unlike any other. This issue of The Journal addresses the handling of medical errors, the termination of mechanical ventilatory support, ethical problems in managed care, and confidentiality issues in the computer era. Guidelines for institutional ethics committees also are presented. These are only a sampling of topics that cut to the heart of bioethics, patient communication, and contemporary medical practice. The more that we study such issues, the more we understand the contributions of medical ethics to medical practice, and the better we serve our patients. PMID- 9401343 TI - The new health care triangle: the ethics of managed care. PMID- 9401344 TI - Electronic health data and the challenges of medical confidentiality. PMID- 9401345 TI - Disconnecting the ventilator: life saving or death delaying? AB - Mechanical ventilators have supported patients with respiratory failure from the era of the polio epidemics to the present. While ventilators can clearly be life saving, many patients receiving this intervention do not survive. In such patients, this treatment is not life saving, but death delaying. Other patients treated with mechanical ventilation may survive, but do not improve to the point where they can become independent of the ventilator. This paper is a review of the prognosis and cost of respiratory failure, ethical issues in removing patients from the ventilator and methods of discontinuing respiratory support. PMID- 9401346 TI - A perspective from clinical and business ethics on adverse events in hospitalized patients. AB - Adverse events occur in a significant, but undetermined, number of hospitalized patients. These types of patient injuries are more often the result of faulty systems than human maleficence. A culture exists among health care providers that discourages the reporting of such events and resists the implementation of formal efforts to eliminate them. This resistance serves to perpetuate the problem. Both business and clinical ethics argue that sound reasons exist for hospitals to reduce, if not eliminate, adverse events. To do so is cost effective, particularly in a managed care environment. It is also at the heart of responsible professional behavior. Physicians are afforded an opportunity to be at the forefront in this quality improvement effort. PMID- 9401347 TI - Synopsis of a practical guide: guidelines for ethics committees. PMID- 9401348 TI - Medicine and money: an approach to compensating university physicians. AB - Academic medical centers have been restructuring faculty compensation plans because decreasing clinical revenues have created budgetary problems. We propose an overall compensation strategy designed to reward high performance of university physicians. The approach incorporates Management by Objectives to improve communication of expectations within the organization and provide college of medicine leadership with much needed mechanisms to align faculty compensation with productivity. PMID- 9401349 TI - Nonstationarity in epileptic EEG and implications for neural dynamics. AB - In this paper, we use a recently developed method to analyze the nonstationarity in time series from intracranial depth and subdural recordings of patients with temporal lobe epilepsy. We show that the nonstationarity in the signal can be accounted for by the variation of a single parameter. We then show that the various dominant nonlinear waveforms observed in different electrodes can be explained by a simple stochastic model in which the mesoscopic collection of neurons, whose potential the electrodes measure, can be on one of two states. The nonstationarity observed in our analysis is a consequence of a time-dependent transition probability between these two states. In general, this transition probability increases as a seizure is approached. The model that we propose incorporates this bistability. We find good agreement between real data and simulated data generated by our model. We understand that this mesoscopic bistability may be associated with the existence of excitation waves traversing the brain in these patients. PMID- 9401350 TI - The effect of community structure on the immunity coverage required to prevent epidemics. AB - Estimation of the immunity coverage required to effectively control disease transmission is an important public health problem. Using data on the eventual size of a major epidemic, we compare estimates based on the simplifying assumption that the community consists of uniformly mixing individuals with estimates obtained when the more complex community structure is acknowledged. The alternative community structures considered include households and localities that are quite separate. Several inequalities are established for estimates of the critical immunity coverage. For several settings, the coverage estimated by assuming an oversimplified community structure is found to actually be an underestimate. A serious consequence of this finding is that we may be misled into believing that we have estimated an immunity coverage that can prevent epidemics when it in fact cannot. The conclusion is that the heterogeneity in the community must be taken into account when estimating the critical immunity coverage. PMID- 9401351 TI - A mathematical model of drug resistance: heterogeneous tumors. AB - A mathematical model is developed to describe the growth and control of a heterogeneous tumor. The main aspect of the model is that it takes into account induced drug resistance. The mathematical model is a system of two ordinary differential equations that describes the growth of the cancer along with the effects of chemotherapy. The model is analyzed to determine what some of the critical parameters are; how we determine an effective treatment; how combination chemotherapy should be delivered; and how this model may help us develop more effective cancer chemotherapeutic treatments. PMID- 9401352 TI - Modeling the covarion hypothesis of nucleotide substitution. AB - A "covarion" model for nucleotide substitution that allows sites to turn "on" and "off" with time was proposed in 1970 by Fitch and Markowitz. It has been argued recently that evidence supports such models over later, alternative models that postulate a static distribution of rates across sites. However, in contrast with these latter well-studied models, little is known about the analytic properties of the former model. Here we analyze a covarion-style model and show (i) how to obtain the evolutionary distance between two species from the expected proportion of sites where two species differ, (ii) that the covarion model gives identical results to a suitably chosen rates-across-sites model if several sequences are compared in pairs by using only the expected proportion of sites at which they differ, (iii) conditions under which the two models will give identical results if the full joint probability matrix is examined, and (iv) that the two models can, in principle, be distinguished when there are at least four monophyletic groups of species. This last result is based on a distance measure that is tree additive under certain versions of the covarion model but, in general, will not be additive under a rates-across-sites model. The measure constructed does not require knowledge of the parameters of the model and so shows that sequences generated by the covarion model do in fact contain information about the underlying tree. PMID- 9401353 TI - Dynamic balance of segregation distortion and selection maintains normal allele sizes at the myotonic dystrophy locus. AB - Myotonic dystrophy (DM), an autosomal dominant neurological disorder, is caused by CTG-repeat expansions at the DMPK locus, with affected individuals having > or = 50 repeats of this trinucleotide. Reduced reproductive fitness of affected individuals and decreased viability of congenital DM have been noted. Expanded CTG-repeat alleles are highly unstable, predominantly yielding even higher repeat sizes. Preferential transmission of longer alleles from heterozygous mothers within the normal size range of alleles also is observed. In view of these observations, it is worth examining how DM has been maintained in human populations for hundreds of generations. We present an analysis of the dynamic properties of a model of joint effects of segregation distortion and selection (intensity of which increases with allele sizes of an individual's genotype). Our mathematical formulation and numerical analyses demonstrate that a weak segregation distortion during female meiosis, together with selection of comparable intensity (within the normal allele size range), can maintain an equilibrium distribution of allele frequencies. Genetic drift, acting in conjunction with the occasional contraction of alleles by mutation, can contribute to the balance of segregation distortion and mutation, in the sense that even weaker selection can explain the observed allele frequencies. The model is applied to CTG-repeat size distributions at the DMPK locus, observed in normal individuals from world populations. PMID- 9401354 TI - Spatially varying equilibria of mechanical models: application to dermal wound contraction. AB - Mechanochemical models based on the Oster-Murray continuum framework have been applied to a variety of biological settings to obtain an understanding of the morphogenesis of living tissues. Wound-healing in mammalian skin is an important example, because a complex sequence of biochemical and biomechanical responses are orchestrated to close a wound by a combination of new tissue formation and wound contraction. Mechanical interactions between dermal fibroblastic cells and the collagen-rich extracellular matrix are crucial in the development of a contracted wound state. We and others have previously proposed mechanochemical models for wound repair to gain a greater understanding of both normal and abnormal healing. In the present work, the existence of spatially varying equilibria of these models is investigated by using a small-stain approximation and phase-plane techniques, with numerical simulations to confirm the analytical predictions. These results are sources of novel insight into the roles of key biological parameters in determining the mechanical properties of a contracted wound. These methods may also be relevant to other morphogenetic scenarios for which similar mechanochemical models have been proposed. PMID- 9401355 TI - Four years experience of primary intra-arterial chemotherapy (PIAC) for locally advanced and recurrent breast cancer. AB - AIMS: To find a means of achieving operability very quickly without the additional discomfort of prolonging systemic chemotherapy. To improve the patient's quality of life by obtaining quick tumor reduction and decreasing systemic toxicity. MATERIALS AND METHODS: From January 1991 to January 1995, 13 patients with locally advanced breast cancer (LABC) and 8 patients with recurrent breast cancer (RBC), were treated by transfemoral Seldinger technique, with the catheter tip placed into the subclavian artery at the basis of the internal mammary artery. The patients received 5-fluorouracil (5FU) 1000 mg, epirubicin (EPI) 30 mg/m2, mitomycin (MMC) 7 mg/m2 over an infusion for 30 minutes. The cycle was repeated every two weeks for three times. RESULTS: The overall response rate was 62%. Stage IIIb and RBC patients had a response rate of 100% and 25% respectively. In respondent patients a measurable response was seen after the first cycle. Ten patients were radically operated. After a media follow-up of 21 months, the overall survival is 52% at 48 months (68% at 48 months and 65% at 34 months for stage IIIb and RBC patients respectively). CONCLUSIONS: PIAC is feasible and effective. In LABC patients it reaches 100% of response rate. Systemic toxicity was absent and the local one was mild. The interval between the starting of PIAC and operation is short. There was an optimal compliance of the patients. PMID- 9401357 TI - Isolated agenesis of the gallbladder. An intraoperative problem. AB - Agenesis of the gallbladder and cystic duct is a rare congenital malformation. In 40-70% of cases this anomaly is associated with other gastrointestinal, skeletal, cardiovascular and genitourinary malformations. Lithiasis of the common bile duct is present in 25-50% of cases. In the majority of cases patients are asymptomatic or have symptoms compatible with a biliary disorder. A preoperative diagnosis is extremely difficult and the absence of the gallbladder is often an intraoperative finding. The authors report a case of isolated agenesis of the gallbladder. The relative embryology, development, diagnostic pitfalls, intraoperative behaviour and therapeutic strategies are discussed. PMID- 9401356 TI - Classical corticosteroids and new lipid peroxidation inhibitors in the therapy of multiple organ failure (MOF). PMID- 9401358 TI - Tuberculin test and tuberculosis control in a low-prevalence country today. PMID- 9401359 TI - Transbronchial lung biopsy: an analysis of 530 cases with reference to the number of samples. AB - To evaluate the diagnostic yield (DY) of transbronchial lung biopsies (TBBs), as the relationship between the DY and the number of tissue specimens taken per TBB, we reviewed the histological and clinical data of 530 consecutive TBBs performed in 516 immunocompetent patients, having either a chronic diffuse lung infiltrate, a localized peripheral lung lesion or hilar adenopathies. The DY (positive TBBs/performed TBBs) varied significantly according to the radiographic pattern and the underlying disease. For chronic diffuse pulmonary infiltrates (n = 244), the overall DY was 50%, but higher figures were obtained for hypersensitivity pneumonitis (92%), sarcoidosis stage II-III (75%), lymphangitic carcinomatosis (68%) and pneumoconiosis (54%). The DY was lower in diffuse tuberculosis (38%) and interstitial pulmonary fibrosis (27%). For localized peripheral lung lesions (n = 205), the overall DY was only 29%, while for sarcoidosis stage I it was 56% (n = 63). Data analysis shows that there is a direct correlation between the number of samples obtained per TBB and the overall DY (i.e. 38% with one to three tissue fragments versus 69% with six to 10, p < 0.01). The increment itself depends on the radiographic pattern and/or the underlying disease which indicates that the probability of diagnostic confirmation per individual tissue sample is not always the same. The clinical implication of these findings is that whereas for some pulmonary diseases the DY is already good with few samples, more samples are to be taken to warrant a satisfactory overall DY. Accordingly, we recommend that at least five to six specimens per TBB should be taken. This number should allow a quite good overall DY in patients with diffuse lung infiltrate. On theoretical grounds, more specimens (seven to 10) should be taken for an optimal DY of localized peripheral lung lesions and of sarcoidosis at stage I. In these indications the clinician should therefore compare the risk-benefit of TBB with a high number of biopsies to the results of other diagnostic procedures. PMID- 9401360 TI - Management of concurrent pleural effusion in patients with lymphoma: thoracoscopy a useful tool in diagnosis and treatment. AB - Pleural effusion represents a frequent feature both of Hodgkin's (HL) and non Hodgkin's (NHL) lymphoma. The aims of the present study were: 1) to analyse the diagnostic accuracy of thoracoscopy as compared to pleural cytology in patients with lymphoma and concurrent pleural effusion; and 2) to evaluate the effectiveness of chemical pleurodesis with the tetracycline derivative, rolitetracycline. Seventeen patients with pleural effusion and concurrent lymphoma (10 NHL and seven HL) were studied. Analysis of pleural fluid revealed the presence of lymphoma cells in six cases (four NHL and two HL); histopathological examination of samples obtained by thoracoscopy was consistent with pleural infiltration by NHL in eight cases and by HL in six cases. Overall sensitivities of pleural cytology and histology were 35 and 82%, respectively. Following chemical pleurodesis, complete response was observed in five of the 17 cases (two NHL and three HL), partial response in four cases (two NHL and two HL), whereas failure was observed in the remaining eight cases. Two patients who had presented failure underwent subsequent pleurectomy by thoracotomy (one case of HL) or video-thoracoscopy (one case of NHL). Complete response was observed in both cases following this treatment. No major complication was recorded after chemical pleurodesis or pleurectomy. Thoracoscopy may be considered a useful tool to evaluate the involvement of pleural space in patients presenting with pleural effusion in the course of lymphoma. Chemical pleurodesis plays an important role in the palliative treatment of this condition. Further studies are necessary to assess the role of pleurectomy in the treatment of such patients. PMID- 9401361 TI - Assessment of the value of fibronectin as a tumour marker in malignant pleural mesothelioma. AB - Fibronectin concentrations both in plasma and pleural effusion were prospectively determined in 60 patients with exudative pleural effusions. Fibronectin concentrations in plasma and pleural fluid in 12 patients with infectious and exudative pleural effusions (mean +/- SD) were 240 +/- 103 and 212 +/- 115 micrograms.mL-1, in 17 patients with primary or metastatic lung carcinoma 242 +/- 104 and 210 +/- 82 micrograms.mL-1, in 13 patients with pleural tuberculosis 246 +/- 77 and 231 +/- 133 micrograms.mL-1, and in 18 patients with confirmed malignant pleural mesothelioma 261 +/- 119 and 276 +/- 188 micrograms.mL-1. There were no significant differences either in the plasma or serum concentrations of fibronectin between groups (p > 0.05). Although pleural fluid fibronectin content appeared to have high specificity (85%), it was found to be an inefficient biological marker for differentiating nonmalignant from malignant pleural effusions due to its low sensitivity (6%). PMID- 9401362 TI - Near fatal asthma and psychopathological characteristics: a group-control study. AB - Psychological factors may play a role in asthma. In particular, emotional upsets have been correlated with fatal asthma attacks, and an abnormal personality attitude is considered to be a risk factor in fatal asthma. Moreover, some authors have recently reported favourable asthma outcome in patients with severe asthma and psychiatric abnormalities, when psychoactive treatment was initiated. On the understanding that people with fatal and "near fatal asthma" (NFA) are components of the same subset of the asthmatic population, we undertook a study aimed at assessing the importance of personality and psychiatric factors in asthma mortality. Between June 1991 and December 1993, a sample of 17 patients with asthma who had experienced one or more near fatal asthma attacks (respiratory arrest, or need for respiratory assistance, or altered conscious state, or arterial carbon dioxide tension (Pa,CO2) > 6.7 kPa (50 mmHg)), and 17 control patients with asthma who had never experienced such an attack (control group) were enrolled. All patients underwent: 1) an interview concerning their personal and family psychiatric history; 2) a psychodiagnostic investigation by a battery of four of the most widely used psychiatric tools: Hamilton scales for anxiety and depression; Zung scales for anxiety and depression; and Minnesota Multiphasic Personality Inventory. No statistical difference was found in psychodiagnostic tests between study and control groups. The psychiatric history was similar in the two groups. Our results suggest that personality characteristics and psychiatric history are not related to asthma outcome, and that the psychiatric approach is not expected to be useful in preventing mortality in asthma. PMID- 9401363 TI - Severe haemorrhagic diathesis in an adult patient with cystic fibrosis after long term antibiotic treatment of pulmonary infection. AB - We describe the case of a 22 yr old male patient with cystic fibrosis, who, after long-term antibiotic treatment of pulmonary infection, developed a haemorrhagic diathesis with severe bleeding from the mucus membrane of the mouth, and haematuria. Rapid recovery was observed after infusion of vitamin K. During 8 months of follow-up, no evidence of recurrence of the clotting disturbances and anaemia were noted. The combination of impaired absorption of vitamin K due to underlying disease with the antibiotic-induced suppression of vitamin K synthesis by intestinal bacteria could be a possible explanation for this disorder. PMID- 9401364 TI - Epithelioid haemangioendothelioma of the lung imitating clinical features of pulmonary histiocytosis X. AB - A young male who was a heavy smoker presented with spontaneous right pneumothorax. A high resolution computed tomography scan showed disseminated nodules up to 1 cm in diameter; the greatest majority of which were sited in the centrilobular zone, though some abutted on the pleural surface. Surgical lung biopsies allowed a diagnosis of epithelioid haemangioendothelioma. The neoplastic tissue infiltrated the wall of bronchioles, partially obliterating them and the visceral pleura. These two histological aspects could be considered as concomitant mechanisms for the appearance of spontaneous pneumothorax. Epithelioid haemangioendothelioma should be added to the list of lung diseases in young heavy smokers that can begin with a spontaneous pneumothorax. PMID- 9401365 TI - Prevalence of tuberculous infection among students in Naples. AB - The aim of this study was to estimate the prevalence of tuberculous (TB) infection in Naples, Italy. The target population was defined as pupils aged 6, 9 and 13 yrs, studying in state and private schools in Naples. A stratified cluster sampling design was used. The stratification criteria were: age (6, 9, 13 yrs); type of school (state or private); and district within the town. Prevalence of TB infection was assessed through a tuberculin Tyne test. Some individual risk factors of tuberculous infection were also investigated by means of a simple questionnaire given to pupils' parents. Among observers reproducibility of test reading was also evaluated. An overall prevalence of TB infection of 5.7% (95% confidence interval (95% CI) 4.6-6.8) was observed in the 1,597 sampled subjects. Skin-test positivity was highly variable with age, ranging from 2.8 (95% CI 1.0 4.6) at 6 yrs to 9.4% (95% CI 7.1-11.7) at 13 yrs. In 458 children (28.7%) response was blindly assessed by three independent observers. Reproducibility of tuberculin skin-test reading was good, with an overall kappa value of 0.718. Only parents' drug abuse was found to be significantly associated with infection. This is the first study to evaluate the prevalence of tuberculous infection in Naples. PMID- 9401366 TI - Pharmacological treatment of exertional dyspnoea in stable COPD patients. AB - Exertional dyspnoea is one of the most common and disabling symptoms in patients with stable chronic obstructive pulmonary disease (COPD). Because little can be done for its resolution, the idea of its symptomatic treatment is attractive. There is no gold standard for the pharmacological management of exertional dyspnoea in stable COPD. A reassessment of the available literature shows the current perspectives and limits. PMID- 9401367 TI - Cystic fibrosis respiratory infections: interactions between bacteria and host defence. AB - In cystic fibrosis, impaired mucociliary clearance leads to chronic endobronchial bacterial infection, mostly caused by Pseudomonas aeruginosa. In the early stage of infection, the pathogen produces several extracellular protein toxins which may contribute to its multifactorial virulence before specific antibodies are produced. P. aeruginosa successfully resists phagocytosis by neutrophils, which dominate the local inflammatory response, by switching from a nonmucoid variant to a mucoid phenotype. Chronic infection and inflammation are characterized by neutrophil-released proteinases which may provide favourable growth conditions for the bacterial opportunist. Aerosol therapy with serine proteinase inhibitors is being investigated in cystic fibrosis. PMID- 9401368 TI - Forced expiratory manoeuvres to increase transport of bronchial mucus: a mechanistic approach. AB - Mucus hypersecretion and impaired mucus clearance are well-known symptoms in patients with chronic obstructive pulmonary disease and cystic fibrosis. These symptoms should not be considered as innocent but they deserve treatment. A well known therapy to improve mucus transport is chest physiotherapy. Forced expiratory manoeuvres are probably the most effective part of chest physiotherapy. Essential in the application of forced expiratory manoeuvres to improve mucus transport is whether airway compression should be used or prevented. The variables that influence the localization and degree of airway compression are expiratory force, lung volume and possibly mouth pressure. With these variables the patient should learn the most efficient form of forced expiration, according to the individual condition of the patient. PMID- 9401369 TI - Aspects of HIV infection: current infection control policies for HIV. AB - A coherent infection control policy within healthcare facilities designed for patients with human immunodeficiency virus (HIV) infection requires the application of a risk management strategy. The central feature is the adoption of universal precautions whereby it is assumed that all patients could potentially be infected by HIV. The major tenets of this are the adoption of good clinical hygiene and the adoption of agreed infection control policies with a consistent approach throughout the institution. This involves a teaching and training programme, and clearly defined policies to protect individuals from HIV infection itself and infection with other pathogens, in particular tuberculosis. Special attention is required for bronchoscopy and lung function, as well as a coherent and proactive policy regarding chemoprophylaxis for HIV infection following accidental injury such as needlestick. PMID- 9401370 TI - Tropical infection and the lung. AB - The term "tropical lung" has been used to describe the lungs that are vulnerable to those indigenous diseases that occur with particular prevalence in tropical countries. International travel, student and cultural exchanges and changing immigration patterns are insidiously transforming the face of medicine in Europe and other developed countries. The climates of tropical countries provide an ideal environment for pathogenic organisms, and their vectors and intermediate hosts, to flourish. In an average out-patient department, 20-40% of patients have been seen with respiratory complaints, and 20-30% of hospital medical admissions are for disorders predominantly affecting the lungs. Pulmonary tuberculosis is common in tropical countries as well as the rest of the world. The other principal environmentally related tropical pulmonary diseases are melioidosis, paragonimiasis, amoebiasis, leptospirosis, gnathostomiasis and tropical eosinophilia. It is essential that the practising clinician be aware of the increasing prevalence of various new and exotic tropical lung diseases. Clinicians in developing countries can now use their clinical skills together with recent developments in immunology, molecular biology and biochemistry to improve the diagnostic accuracy and therapeutic effectiveness related to tropical lung infections. Treatment, if inappropriate, may not only be worthless but, in many cases, extremely harmful and even fatal. PMID- 9401371 TI - Inspiratory muscle dysfunction as a cause of death in COPD patients. AB - Although fatigue of the inspiratory muscles has been well documented, its prevalence in patients with chronic obstructive pulmonary disease (COPD) and its influence on mortality are unknown, because of the lack of a simple, clinically available diagnostic test. The hypothesis and experimental evidence relating inspiratory muscle dysfunction to the development of hypercapnia and hypercapnic ventilatory failure are reviewed. Since a poor prognosis in COPD is associated with carbon dioxide retention, inspiratory muscle weakness and/or fatigue may have an association with survival in these patients. PMID- 9401372 TI - How women and youth are targeted by the tobacco industry. AB - Women and youth are fast becoming casualties of the cigarette epidemic. Although global male smoking rates are almost four times higher than female rates (47 versus 12%), male rates are declining, while female smoking rates are increasing. With the significant decline in tobacco consumption over the past two decades in Western countries, the tobacco industry has responded by developing new markets. The main targets of the tobacco industry are women and youth: in developing countries, in lower socioeconomic groups, and those with independent careers. The tobacco industry recognizes that women and youth represent a major untapped market and are targeting them with aggressive advertising, marketing and promotional campaigns, sponsorship, and employment in the growing tobacco industry. In addition, they are applying political and commercial pressure to open new markets in developing countries. These initiatives of the tobacco industry have resulted in an increase in tobacco use among women and youth. The economic arguments put forward by the tobacco industry of providing employment for many people have to be balanced against the social and financial drain that tobacco costs the economy in the health area, and the human suffering from preventable diseases resulting from smoking. PMID- 9401373 TI - Influence of a secondary genetic factor in cystic fibrosis genotype-phenotype correlations. PMID- 9401374 TI - Bronchoscopic biopsy techniques in the diagnosis and staging of lung cancer. AB - In a review of the bronchoscopic biopsy techniques currently available for the diagnosis and staging of lung cancer, the author individually analyses the sampling instruments and procedures used in cancer of the central airways, peripheral lung lesions, and in the study of hilar and mediastinal lymph nodes. With regard to central bronchial lesions, data concerning diagnostic yield, the advantages and limits of forceps biopsy, brushing, washing and transbronchial needle aspiration are reported. With the integration of two or more of the sampling methods, a cytohistological diagnosis can be obtained in almost all cases. In the field of peripheral lung cancer, the diagnostic possibilities of the transbronchial approach are described. The experience of the Regional Hospital of Ancona, Italy on 1,027 patients affected by peripheral pulmonary nodules or masses is reported. The biopsy technique is based on the integration of transbronchial (using forceps biopsy and transbronchial needle aspiration) and percutaneous approaches, and on a team approach with the close co-operation of a pulmonologist, a radiologist and a cytopathologist, all simultaneously present in the diagnostic suite during the procedures. On the basis of the results obtained, the author suggests that, in peripheral pulmonary lesions, the transbronchial approach should generally be performed before the percutaneous needle aspirate, especially in patients who are candidates for surgery. The transbronchial approach has the advantages of allowing an examination of the tracheobronchial tree and staging of lymph nodes with a lower incidence of complications. In addition, the diagnostic yield of transbronchial needle aspiration in the study of hilar and mediastinal lymph nodes is analysed. This method, if positive, plays a major role in the staging of lung cancer and makes it possible to avoid unnecessary surgical procedures. Knowledge of the advantages and limits of different sampling instruments and procedures, and of their integration, is essential to optimize the diagnostic management of each patient with lung cancer. The goal is to maximize the diagnostic possibilities whilst minimizing risk and reducing costs. PMID- 9401375 TI - Improved simulation system for routine cardiopulmonary exercise test equipment. Part II: A new metabolic simulator system: practical applicability. The European Coal and Steel Community (ECSC) Working Group on Standardization of Stress-test Methods. AB - The utilization in clinical practice of an improved metabolic calibrator was tested. The morphological characteristics and integration patterns of the signals were checked with a reference stress-test instrument and compared with physiological recordings. No significant differences were observed. Conversion formulae were developed to compare the calibrator's data with those produced by routine instruments. Simultaneous graphical representations of erroneous and reference signals and computer processing of data allowed on-line detection of even minimal sources of error in dynamic working conditions. PMID- 9401376 TI - Vaccination and the eradication of infectious diseases. PMID- 9401377 TI - Unreported trials: an opportunity. PMID- 9401378 TI - Comparison of methods to assess coronary heart disease prevalence in South Asians. AB - BACKGROUND: Migrants from the Indian subcontinent (South Asian migrants) in the United Kingdom have high mortality from coronary heart disease (CHD) in comparison to the indigenous population. Few studies have assessed the prevalence of CHD in South Asians, and the applicability of conventional survey methods in this population is not known. In this pilot random population survey of South Asian men and women living in West London, the prevalence of CHD as judged by the Rose questionnaire, past cardiac history, cardiologist and resting electrocardiogram were compared. METHODS: Subjects aged 30-64 years from randomly selected households were invited for a cardiological assessment. A lay person administered the Rose questionnaire and recorded the past cardiac history. A cardiologist also made an independent assessment and a 12-lead electrocardiogram was recorded and analysed according to the Minnesota code. RESULTS: Three hundred and seventy-six individuals (192 men and 184 women) were assessed. The prevalence of angina in men and women, respectively, was 3.1% and 4.9% by the Rose questionnaire; 2.6% and 2.2% by past cardiac history; and 4.2% and 0.5% according to the cardiologist. The prevalence of myocardial infarction in men and women, respectively, was 5.2% and 2.2% by the Rose questionnaire, 3.6% and zero by past cardiac history and 3.6% and 0.5% by the cardiologist. Q/QS codes were present in 1.6% men and 0.5% women and ischaemic codes in 13% men and 14% women. Ischaemic changes were not associated with any cardiac history in 72% of men and 92% of women. For a diagnosis of CHD in men, there was poor agreement between the Rose questionnaire and either the past cardiac history or the cardiologist's assessment, but moderate agreement between the past cardiac history and the cardiologist. Agreement was poor between all three methods for a positive diagnosis of CHD in women. CONCLUSION: Current accepted epidemiological methods for assessing CHD prevalence may be inaccurate in South Asians, especially women. Electrocardiogram abnormalities suggestive of ischaemia are common in South Asians and are usually not associated with evidence of CHD. Thus, their value as indicators of CHD is questionable. PMID- 9401379 TI - Modified Glasgow Coma Scale to predict mortality in children with acute infections of the central nervous system. AB - BACKGROUND: This study aimed to identify the predictors of hospital mortality in children with acute infective disorders of the central nervous system using an aggregate Modified Glasgow Coma Scale (MGCS) score and other clinical variables assessed within 24 hours of hospitalization. METHODS: We did a prospective cohort study in a teaching and referral hospital in Lucknow, North India. Consecutive children aged 1 month to 12 years of age admitted with acute infective disorders of the central nervous system were included in the study. The diagnosis was based on the presence of symptoms of fever, headache or irritability with or without vomiting, and either altered sensorium or first episode of seizures or both. The main outcome measure was hospital-based mortality. RESULTS: Of the 230 patients included in the study, 42.2% had pyogenic meningitis, 36.9% had tuberculous basal meningitis and 20.9% had meningo-encephalitis. There were 43 (18.7%) deaths of which 44.2% were within 3 days of admission. Death was associated with the day 1 aggregate MGCS score only. The area under the curve of four strata of aggregate MGCS was 0.63 (SE 0.05). The likelihood ratio for discharge with an aggregate MGCS score of < 5 was 0.52 (95% CI:0.29-0.95) and > 10 was 5.52 (9% CI:1.02 31.96). CONCLUSION: The MGCS can be used to predict discharge in patients with acute infective disorders of the central nervous system within 24 hours of hospitalization. The scale is simple, can be applied at the bedside and does not depend on any investigations. In developing countries with limited investigative facilities it can be used for identification and selective referral of patients with a higher risk of death to specialized centres. This study validates the predictive value of the MGCS. PMID- 9401381 TI - Microbial resistance to drugs--a universal problem in urgent need of a comprehensive approach. AB - The last two decades have seen an increase in bacterial resistance to commonly used antibiotics all over the world. In the past five years the emergence of vancomycin-resistant Enterococcus and multidrug-resistant Streptococcus pneumoniae was particularly notable. Several factors have contributed to this, including inappropriate use of readily available antibiotics, survival of high risk patients in critical care units, burn wards and cancer centres following treatment with multiple antibiotic combinations, increasing poverty and worsening of living conditions. The data available from South-east Asia, albeit limited, indicate that prevalence of antibiotic resistance in bacteria isolated from human and animal sources is higher than that reported from western countries. To combat this global problem, a multi-pronged approach is needed. Accurate antibiotic susceptibility data will be required to define the extent of the problem. Medical experts and scientific organizations will have to develop guidelines for the use of antibiotics in ambulatory, inpatient and animal husbandary areas. Cooperation between the medical community, regulating agencies and pharmaceutical industry will be needed to define policies governing the sale of antibiotics, drug promotional materials, physician education programmes and consumer education regarding the hazards of inappropriate antibiotic use. For long term control of infectious diseases, it is imperative that existing vaccines be appropriately utilized and new vaccines developed. PMID- 9401380 TI - Distribution of vascular lesions in ischaemic stroke: a magnetic resonance angiographic study. AB - BACKGROUND: Carotid endarterectomy is now an accepted modality for reducing the threat of recurrence of ischaemic strokes in patients with severe carotid artery stenosis. However, the incidence of carotid artery stenosis, and hence the applicability of carotid endarterectomy in the Indian population is not known. We conducted a prospective study to detect and quantify extracranial and intracranial arterial lesions using magnetic resonance angiography in consecutive patients with ischaemic strokes. METHODS: All patients with recent onset of ischaemic stroke (< 4 months) had a magnetic resonance angiography done to evaluate the neck vessels as well as the circle of Willis and its branches. The degree of stenosis of the internal carotid or common carotid artery was measured according to the criteria described by the North American Symptomatic Carotid Endarterectomy Trial (NASCET) collaborators. The site and extent of the extracranial and intracranial arterial lesions were correlated with the clinical features and the pattern of infarcts on magnetic resonance imaging or computerized tomographic scan of the brain. RESULTS: The magnetic resonance angiography was abnormal in 56 out of 100 patients included in the study. Severe stenosis (> 70%) of the extracranial carotid arteries was seen in 26 patients. Lesions suitable for carotid endarterectomy were present in only 11 patients (42.3% of those with severe stenosis). CONCLUSION: Our results are in contrast to those reported from western countries where the likelihood of a surgically correctable lesion being present is 60%-70%. We found operable lesions in only 11%. Intracranial atherosclerotic disease causing strokes is probably more common in India. Therefore, although carotid endarterectomy is the only accepted surgical procedure for secondary prophylaxis of stroke, there is a need to find an alternative surgical intervention for the predominantly intracranial pathology found in the Indian population. PMID- 9401382 TI - Gender differences in depression in relation to life events. PMID- 9401383 TI - The prognosis for very tiny neonates. PMID- 9401384 TI - The enigma of pituitary abscess. PMID- 9401385 TI - Chest pain. PMID- 9401386 TI - Ageing. PMID- 9401388 TI - Making an ass of the law. PMID- 9401387 TI - Multimedia in health. PMID- 9401389 TI - The Scottish Health Survey--alcohol consumption in Scotland. PMID- 9401391 TI - Armed Forces Medical College, Pune. PMID- 9401390 TI - How can we help today's youth? PMID- 9401392 TI - Research methods in cancer: clinical trials, epidemiology and audit, Tata Memorial Hospital, Mumbai, 28 February and 1 March 1997. PMID- 9401393 TI - Diphtheritic conjunctivitis. PMID- 9401395 TI - Health services for India's urban poor. PMID- 9401394 TI - Susceptibility of Staphylococcus aureus at a rural clinic. PMID- 9401396 TI - Chemical injury due to colours used at the festival of Holi. PMID- 9401397 TI - A new method for quantifying the cognitive impairment. Its application in patients with vascular dementia. AB - Seven-three patients diagnosed with vascular dementia (VD) left to their natural evolution have been observed during a period of time of one year. The objective of this study is to find an index to quantify the degree of loss of cognitive capabilities in patients affected by VD, as well as to define the minimum loss of punctuation in this index, that can only be caused by an unfavourable evolution of the disease. This index, which we call Percentual Variation Index (PVI), is based on the evolution of the punctuations of the Cognoscitive Mini-Exam (CME) after a follow-up of one year. Our definition states that in order to consider patients as having cognitive impairment of vascular origin, they must lose more than 10% of their basal CME score in one year. This method could be useful in assessing the therapeutic efficacy of drugs used in treating such illnesses. PMID- 9401398 TI - Intracranial meningioma recognition following general anesthesia: a surprisingly acute form of presentation. PMID- 9401399 TI - Diurnal enuresis. AB - Daytime wetting is a common problem in childhood; in most cases, it is an intermittent or self-limited problem with a benign and easily identifiable cause. The history is the most important aspect of the assessment. A urinalysis is helpful to look for UTI, and ultrasonography of both the kidney and the bladder is an excellent noninvasive screening study. More invasive diagnostic imaging studies are necessary only if the patient has symptoms or signs suggestive of urethral obstruction, neurogenic bladder, Hinman syndrome, or ectopic ureter. With time and appropriate treatment, the prognosis is excellent for the majority of patients. PMID- 9401400 TI - Child occupant protection in motor vehicles. PMID- 9401401 TI - What's new in pediatric emergency medicine. PMID- 9401402 TI - Musculoskeletal injury in children. PMID- 9401403 TI - Tobacco. PMID- 9401404 TI - Poor growth after cranial irradiation. PMID- 9401405 TI - Is fever bad? PMID- 9401406 TI - Role of Ito cells in the liver function. AB - Ito cells (perisinusoidal fat-storing cells, stellate cells, lipocytes) of the liver are mesenchymal cells located in the space of Disse. They are the main place of vitamin A storage in characteristic lipid droplets. Moreover, they demonstrate synthetic activity of collagens and other extracellular matrix proteins involved in hepatic fibrosis. Their long, contractile cytoplasmic processes encompass the sinusoids and can affect sinusoidal blood flow. Because of their location in the space of Disse and connection with other sinusoidal and parenchymal cells, they are probably involved in local neurotransmission and paracrine regulation of numerous liver functions. As a source of cytokines, prostaglandins and other bioactive substances, produced in the response to noxious agents, they play a crucial role in the mechanisms of liver injury, regeneration and fibrosis. This article is a review of the current state of knowledge on Ito cells structure and function. PMID- 9401407 TI - Liver cell dysplasia in liver cirrhosis and hepatocellular carcinoma. AB - The aim of the study was to assess the incidence of both types of liver cell dysplasia and concomitance with cirrhosis, hepatocellular carcinoma (HCC) and positive reaction for HBsAg in the autopsy material and an attempt to determine a relationship between these two types of liver cell dysplasia and hepatocellular carcinoma. Autopsy material included 102 cases of hepatocellular carcinoma, 101 cases of hepatocirrhosis without accompanying cancer and 106 control cases. Histological specimens stained with HE were analyzed for the presence of large liver cell dysplasia (LLCD) according to Anthony et al., small liver cell dysplasia (SLCD) according to Watanabe et al., the presence of macroregenerative nodules (< 8 mm) and antigen HBs (stained with orcein according to Shikata). The detected LLCD were also assessed semiquantitatively taking into account the number of dysplastic areas in a given case. Statistical significance of the results was tested with the chi square test. LLCD was most frequently detected in HCC with concomitant cirrhosis (55.3%), then in cirrhosis without HCC (40.6%), and in HCC without cirrhosis only in 12.5%. LLCD was found significantly more frequently (p < 0.05) in cirrhosis with HCC than in cirrhosis without HCC. Antigen HBs was found in 25.6% of cirrhoses and/or HCC. No significant differences in the presence of HBsAg were seen between the analyzed groups. The incidence of LLCD and HBsAg in controls was significantly lower than in other groups. A mean age at death in case of cirrhosis with HCC subdivided into that with or without LLCD was not significantly different, whereas in case with cirrhosis with LLCD age at death was 10.8 years higher (the difference statistically significant). Analysis of material with respect to gender revealed a high proportion of men in case of HCC with concomitant cirrhosis but without LLCD (13:1). A strong relationship was seen between the presence of positive reaction for HBsAg and LLCD (p < 0.001). Also the intensity of LLCD positively correlated with the presence of HBsAg. Furthermore, a positive correlation was seen between the presence of LLCD and macronodular cirrhosis (posthepatitic). The present findings suggest a closer relation between HBV infection and LLCD than between cirrhosis or HCC and LLCD. Also morphological patterns of LLCD foci do not confirm the hypothesis of some investigators about the precancerous character of these lesions. In the whole current material only seven cases of SLCD were detected. They were all present in cirrhotic livers with concomitant HCC. Both the morphological pattern of these lesions and their sometimes discerned close spatial relation with HCC foci indicate that SLCD is an alternative way of HCC development. PMID- 9401408 TI - Gastric fundic gland polyps (Elster's polyps) and Helicobacter pylori-induced gastritis. AB - In a series of 555 gastric polyps characteristic Elster's polyps were identified in 31 cases. Spiral bacteria (Helicobacter pylori) in these polyps were sporadic, much less frequent (9.7%) than in hyperplastic polyps (35%) in the present series and in relation to the bacteria frequency found in our randomly chosen gastric biopsy specimens (43%). The present results indicate that Elster's polyps are not readily colonized by Helicobacter pylori and accordingly, they do not show the signs of active gastritis. The reasons for this are unknown. One of the mechanisms preventing from bacterial colonization may be a different from normal gastric mucosa and other polyps character of mucus produced by glandular neck cells, which was found in our series of gastric fundic gland polyps. PMID- 9401409 TI - Myelinated axon undergoes complete demyelination in the panencephalopathic--but it is merely subjected to the Wallerian degeneration in the polioencephalopathic type of transmissible spongiform encephalopathies. AB - We report here on axon and myelin changes in panencephalopathic type of Creutzfeldt-Jakob disease as opposed to polioencephalopathic models of scrapie. In CJD, myelinated axons presented various pathological changes. Initially the myelin sheath was separated by cytoplasmic tongues into several concentric bands. Cellular processes penetrated between layers of myelin and lifted away the outermost lamellae. Then a complicated labyrinth of concentric cellular processes, clearly belonging to either astrocytes or macrophages invested myelinated axons. In terminal stages axons completely denuded of myelin were seen in the centre of a concentric network of cellular processes or spirals of myelin were seen surrounded by such processes. The myelin fragments penetrated into astrocytes or macrophages where they underwent final digestion. Macrophages containing fragments of axons, paracrystalline lamellar bodies and myelin debris were abundant in this model. In two models of scrapie myelin dilatation, not unlike that seen in the panencephalopathic type of CJD was observed. Several altered axons were also seen, but these presented only typical features of Wallerian degeneration. Even the most damaged fibres consisted of "empty" vacuoles (the axon itself was lost) surrounded by a narrow rim of oligodendroglial cytoplasm. Thus, myelinated fibre degeneration in this polioencephalopathic model merely represents the sequel to preceding neuronal degeneration. In conclusion, it seems that myelinated fibres are eventually denuded of myelin in the panencephalopathic type of CJD but undergo Wallerian degeneration in polioencephalopathic types of scrapie. Because myelin dilatation is observed in both pan- and polioencephalopathic type of TSE and because their formation is probably mediated by cytokines released from astrocytes and microglia, we hypothesised that this mechanism operates early in the fibre destruction and must be supplemented later in the course of the disease by other, currently unknown mechanisms. PMID- 9401410 TI - Primary IgA nephropathy. Contrasting morphometric insight into glomerular and interstitial changes in younger and elderly patients. AB - Twelve patients with primary diffuse IgA nephropathy (IgAN) aged under 45 years (IgAN < 45), and eleven patients with this glomerulopathy aged over 45 years (IgAN > 45) for whom both light and electron microscopy as well as immunofluorescence microscopy and full clinical data were available were examined quantitatively and compared with fifteen normal controls. Morphometric investigations were performed by means of a computer image analysis system to characterize quantitatively IgAN in patients over and below 45 years of age as well as to study whether changes in quantitatively analyzed glomeruli could correlate with hematuria, which is thought as the main renal symptom in this glomerulopathy. The study revealed that the mean values of total glomerular cells per total glomerular area, total glomerular cells per unit of glomerular area, mesangium (% of total glomerular area) and relative interstitial volume were in both IgAN < 45 and IgAN > 45 groups increased in comparison with normal controls, most of them significantly. The comparison of these parameters between IgAN < 45 and IgAN > 45 groups showed that the mean values of total glomerular cells per total glomerular area and total glomerular cells per unit of glomerular area were significantly greater in IgAN < 45 group, but relative interstitial volume was significantly increased in IgAN > 45 patients. Moreover, there were significant positive correlations between total glomerular cells per unit of glomerular area and hematuria, but not between glomerular mesangium (% of total glomerular area) and hematuria in both IgAN < 45 and IgAN > 45 groups. In conclusion, we found evident quantitative differences between glomerular and interstitial parameters in younger and elderly patients. Although close relationship was observed between glomerular hypercellularity and the degree of hematuria in both younger and elder patients, this association is not clear and its pathogenesis remains to be shown. PMID- 9401411 TI - Tumor necrosis factor-alpha induces emphysema-like pulmonary tissue rebuilding. Changes in type II alveolar epithelial cells. AB - The effect of repeated doses of TNF-alpha on the histological picture of the pulmonary tissue was analyzed in the present study. Special attention was paid to the lung rebuilding processes. TNF-alpha was applied intraperitoneally for two weeks in a dose of 10 micrograms/0.5 ml PBS/24h. Morphological analysis of the pulmonary tissue was performed after 1 and 28 days following the last TNF-alpha dose. The study revealed focal pulmonary tissue rebuilding with emphysema-like changes twenty eight days following termination of TNF-alpha administration. The rebuilding processes included interalveolar septal atrophy, collagen accumulation and damage-repair changes in type II alveolar epithelial cells. It has been demonstrated that apart from the protease-antiprotease hypothesis of the lung emphysema, the inflammatory-repair hypothesis should be considered. Both hypotheses are complementary to each other and interpret the emphysema-like changes as complications of various pathological conditions of the pulmonary tissue. PMID- 9401412 TI - Ultrastructural study of subependymal giant cell astrocytoma: unusual paracrystalloid inclusions in tumour cells. AB - Subependymal giant cell astrocytoma (SEGA) is a tumour with a broad range of morphological, immunohistochemical and ultrastructural neoplastic cellular features. We report here the presence of this tumor in two 7-year-old-boys presented with a lateral intraventricular mass. Further clinical investigation confirmed the diagnosis of tuberous sclerosis. Ultrastructural and immunohistochemical investigation revealed that glial differentiation was more pronounced in these cases. Interestingly, tumour cells containing unusual paracrystalline inclusions, infrequently described in SEGA, were identified ultrastructurally. These inclusions are probably not related to other cellular organelles. PMID- 9401413 TI - A case of mitochondrial myopathy with MELAS-like features and polyneuropathy: ultrastructural and molecular studies. PMID- 9401414 TI - Multiple fundic gland polyps in duodenal mucosa not associated with colonic polyposis. AB - One of extracolonic manifestations of Gardner's syndrome and familial adenomatous polyposis (FAP) are fundic gland polyps (FGP) located typically in the gastric body. Rarely FGP develop in the absence of intestinal polyposis. A case of FGP in the duodenal bulb without associated pathological changes in the large bowel has been reported. PMID- 9401415 TI - Primary malignant melanoma of the esophagus. A case report. AB - A case of primary malignant melanoma of the esophagus has been reported. This rare entity accounts for less than 0.2% of all esophageal neoplasms. A polypoid, deep blue and tough tumour was present on endoscopic examination during which a few samples were taken. Microscopically melanotic tissue occupied the whole submucosa and scanty deposits of melanin pigment were dispersed. Positive immunohistochemical stain with HMB-45 antibody confirmed the diagnosis of malignant melanoma. The patient was admitted to the Oncological Center for resection of the tumour, which is the treatment of choice in such cases. PMID- 9401416 TI - Iron crosses the endosomal membrane by a carrier-mediated process. PMID- 9401417 TI - Whole-heart modeling: progress, principles and applications. PMID- 9401418 TI - alpha-Amylase family: molecular biology and evolution. PMID- 9401419 TI - Heart rate variability: origins, methods, and interpretive caveats. AB - Components of heart rate variability have attracted considerable attention in psychology and medicine and have become important dependent measures in psychophysiology and behavioral medicine. Quantification and interpretation of heart rate variability, however, remain complex issues and are fraught with pitfalls. The present report (a) examines the physiological origins and mechanisms of heart rate variability, (b) considers quantitative approaches to measurement, and (c) highlights important caveats in the interpretation of heart rate variability. Summary guidelines for research in this area are outlined, and suggestions and prospects for future developments are considered. PMID- 9401420 TI - Cardiovascular recovery from stress and hypertension risk factors: a meta analytic review. AB - Recent research has suggested that cardiovascular recovery from stress can play a potential role in hypertension pathogenesis. Sixty-nine studies were included in a meta-analytic review to evaluate the effect of various hypertension risk factors (e.g., race, lack of exercise) on cardiovascular recovery from stress. Small mean effect sizes were observed for studies examining hypertension status and race as risk factors associated with delayed diastolic blood pressure recovery. Lack of fitness was also associated with delayed heart rate recovery. These results revealed that, for the specified risk factors and cardiovascular variables, high-risk individuals exhibited delayed cardiovascular recovery as compared with low-risk individuals. Further, the relationships between hypertension status, race, and cardiovascular recovery were typically associated with the use of "active" laboratory stressors. The relationship between lack of fitness and cardiovascular recovery was also associated with the use of "active" and exercise laboratory stressors. PMID- 9401421 TI - When syntax meets semantics. AB - Three experiments concerning the processing of syntactic and semantic violations were conducted. Event-related potentials (ERPs) showed that semantic violations elicited an N400 response, whereas syntactic violations elicited two early negativities (150 and 350 ms) and a P600 response. No interaction between the semantic and early syntactic ERP effects was found. Sentence complexity and violation probability (25% vs. 75%) affected only the P600 and not the early negativities. The probability effect was taken as evidence that the P600 resembles the P3b. The temporal order of word processing in a sentence as suggested by the data was such that a more automatic syntactic analysis was performed (earlier syntactic-related negativities) in parallel with a semantic analysis (N400), after which a syntactic reanalysis was performed (P600). A reanalysis interpretation of the P600 could explain why the extent of the reanalysis differed with syntactic complexity and probability of ungrammaticality. PMID- 9401422 TI - Development and topography of auditory event-related potentials (ERPs): mismatch and processing negativity in individuals 8-22 years of age. AB - How do event-related potentials (ERPs) reflecting auditory processing develop across adolescence? Such development was described for five ERP components in four groups of 11 healthy participants with mean ages of 10, 14, 17, and 21 years. Data from 19 sites during diffuse (passive) and focused (discrimination) attention in a three-tone oddball were analyzed to see how ERP loci varied with age for tone type, attention condition, and for four types of difference waves reflecting nontarget and target comparisons. Age interacted with site for most components. P1 loci sensitive to rare tones moved posteriorly and N1 loci lost their right bias in early puberty. The P2 loci did not move anterior to Cz until adulthood. N2 amplitude, sensitive to attention condition, developed a frontal focus by 17 years. Right-biased P3 loci moved to the midline with focused attention similarly in all age groups. Difference waves developed in three stages: In 10-year-old participants, early deflections (< 150 ms) were diffusely distributed; in midadolescent participants, the main frontal negative component (150-300 ms) became well formed and lost an earlier right bias; and for participants 17 years old and older, the late positive complex developed a right bias in target-derived waves. Latency decreases for early frontal components were marked in participants 10-14 years old and for later posterior components in participants 14-17 years old. Major developments appeared at the onset of adolescence in early stimulus selection processes and during adolescence in the differential use of this information (N2- and P3-like latencies). PMID- 9401423 TI - Mismatch negativity during objective and subjective sleepiness. AB - The mismatch negativity (MMN) and P3 of auditory event-related potentials were studied during subjectively and objectively (physiologically) defined sleepiness under optimal stimulus conditions for MMN elicitation. The MMN and P3 were elicited by either small or large unattended auditory deviants presented to the left ear. The participant's task was to detect either rare auditory targets presented to the right ear or rare changes in the light flashes. Eleven young adults served as participants in a nighttime experiment. The MMN declined especially at Fz and Cz but not so markedly at the right mastoid as either subjective or objective alertness decreased. The amplitude of P3 also decreased during sleepiness. The attenuation of the MMN was paralleled by a decline in behavioral performance. The results show that the MMN is attenuated by a decrease in alertness even before an actual sleep state is reached. PMID- 9401424 TI - Effects of attentional and stressor manipulations on the P50 gating response. AB - The decline in amplitude of the P50 component of the event-related potential to the second of paired clicks has been suggested as a measure of preattentional gating. Two experiments were conducted to assess the effects of attention and a psychological stressor on P50. Experiment 1 included two choice reaction time tasks designed to direct attention selectively to the first or second click in each pair. Results suggest that the N100 component was responsive to attentional manipulations, whereas P50 was not affected. Experiment 2 examined the impact of a brief psychological stressor on the P50 response. Parallel mental arithmetic tasks were administered silently and orally. Self-report and measures of autonomic activity were used to assess the level of stress occurring during the performance of the mental arithmetic tasks. Results indicate that P50 suppression was sensitive to the acute stressor, the oral mental arithmetic task. Implications of these findings for studies of P50 suppression in schizophrenia are discussed. PMID- 9401425 TI - Storage of feature conjunctions in transient auditory memory. AB - The purpose of this study was to determine whether feature conjunctions are stored in transient auditory memory. The mismatch negativity (MMN), an event related potential that is elicited by stimuli that differ from a series of preceding stimuli, was used in this endeavour. A tone that differed from the preceding series of stimuli in the conjunction of two of its features, both present in preceding stimuli but in different combinations, was found to elicit the MMN. The data are interpreted to indicate that information about the conjunction of features is stored in the memory. PMID- 9401426 TI - An in vivo comparison of two circumferential penile strain gauges: the introduction of a new calibration method. AB - This study assessed the comparability of two types of penile strain gauges using both a circular and a newly developed oval calibration device. A group of 25 sexually functional participants placed both an electromechanical and an indium/gallium-in-rubber strain gauge on the penis and viewed an erotic film. The electromechanical gauge as calibrated on the circular device resulted in greater penile circumference changes than the indium/gallium-in-rubber gauge. Mean circumference changes were not different for the two strain gauges when the oval calibration device was used. The use of an oval calibration device improves ecological validity of calibration of penile strain gauges. Standard inclusion of this method in studies on male sexual response will increase comparability of research findings. PMID- 9401427 TI - Suppression of sleepiness in drivers: combination of caffeine with a short nap. AB - Previous research has shown that caffeine and a < 15-min nap effectively and separately reduce sleepiness in drivers for 1 hr. In the present study, we examined in 12 sleepy individuals the treatments combined, taken during a 30-min break, prior to a longer (2 hr) continuous monotonous afternoon drive in a car simulator. Nonnap comparisons were 200 mg caffeine only and placebo. For placebo, driving incidents, subjective and electroencephalographic measures of sleepiness all reflected a mid-afternoon peak. This peak was significantly reduced by caffeine and eliminated by the combined treatment, which reduced incidents to 9% of placebo levels versus 34% of placebo levels for caffeine alone. Naps comprising "nonsleep dozing" were still effective. PMID- 9401428 TI - Startle reflex modulation by pleasant and unpleasant odors in a between-subjects design. AB - Participants were presented with an acoustic startle probe while they smelled either a pleasant odor or an unpleasant odor (n = 40 per condition). Within each condition, participants were presented with four blocks in which odor was present and four in which odor was not present (no-odor control). Significant differences were found in both conditions between the odor and no-odor blocks. In the unpleasant odor condition, blink magnitude was greater during the odor blocks; in the pleasant odor condition, blink magnitude was smaller during the odor blocks. PMID- 9401429 TI - Interaction of the von Willebrand factor with platelets and thrombosis. AB - The human von Willebrand factor (vWf) is a multimeric glycoprotein present in plasma, platelets, endothelial cells and subendothelium and synthesized in endothelial cells and megakaryocytes. vWf plays a pivotal role in the mechanisms of blood clotting and platelet thrombus formation; quantitative and qualitative abnormalities of vWf cause the most common congenital bleeding disorder in man, the von Willebrand disease. vWf stabilizes factor VIII and interacts with subendothelial components and with platelet membrane receptors. The multimeric structure of vWf provides an array of binding sites which allows multivalent interactions with its ligands, thus supporting the formation of stable platelet aggregates at the site of vascular injury, particularly under flow conditions characterized by high shear stress. In the last years, remarkable progress has been made toward understanding the structure of vWf protein and gene, and the elucidation of many structure-function relationships, which may result in improved therapeutic intervention for vWD patients, and in the development of effective strategies for antithrombotic therapy. PMID- 9401430 TI - Acute intravascular haemolysis with massive microspherocytosis in a 75-year-old woman. AB - Acute intravascular haemolysis (AIH) sometimes occurs in patients with sepsis or bacteraemia, mainly due to clostridia or Salmonella sp., and may be a life threatening condition. We describe a case of AIH in a 75-yr-old woman with chronic cholelithiasis. Blood and stool cultures were repeatedly negative, but the massive microspherocytosis, typically observed in clostridia infections, oriented our diagnosis. The patient was treated with antibiotics and for a rapid worsening of her conditions, which could have led to the onset of a multi-organ dysfunction syndrome (MODS), with plasma exchange and, subsequently, haemodialysis, with satisfactory results. PMID- 9401431 TI - Influence of alcohol consumption on the initial development of chronic pancreatitis. AB - The aim of our study was to analyze the influence of alcohol consumption on the early clinical manifestations of alcoholic chronic pancreatitis of the 517 patients in whom chronic pancreatitis was initially suspected, 158 were diagnosed with this disease; of these, alcohol was considered the cause in 136 (86.1%). Alcohol was considered a major etiologic factor when mean consumption was > or = 60 grams per day for at least 4 years. Alcohol consumption, initial clinical manifestations and time of onset were considered up until the moment of diagnosis in all patients. The sex distribution was 133 men (97.8%) and 3 women (2.2%). The average age was 22 +/- 6.5 years at onset of alcoholism, 38 +/- 9.4 years at onset of clinical features, and 44 +/- 9.4 years at diagnosis. The interval between the onset of alcoholism and the initial clinical manifestations was 15.8 +/- 8.8 years, and the interval between the latter and diagnosis was 6.1 +/- 4.9 years. Average alcohol consumption was 162 +/- 8 grams/day and total consumption was 1312 +/- 1017 kg. A statistically significant relationship was found only for mean alcohol consumption and abdominal pain. We found a higher frequency of acute pancreatitis outbreaks, calcifications, steatorrhea and diabetes until the moment of diagnosis in the higher alcohol consumption groups, although the relationship was not statistically significant. PMID- 9401432 TI - Laparoscopic gastroplasty. Technique and preliminary results in patients with morbid obesity. AB - Morbid obesity is an aesthetic, social and psychological problem, with characteristic morbidity and mortality. Surgical treatment has been developed due to the high failure rate of medical treatment. In our previous experience with 40 patients, open transverse gastroplasty reinforced with mesh, has been found to be a good procedure to deal with this problem, with very few postoperative complications. In March 1995 we started a prospective study to evaluate the feasibility and results of the same technique done laparoscopically. We operated on 10 morbid obesity patients with several failed medical treatments. Laparoscopic surgery was felt to be easier and faster than the open technique. Weight loss was similar to that in our previous patients with open surgery, but postoperative pain and complications were less frequent, and the aesthetic results were better. More patients and a longer follow-up will be needed to provide definitive conclusions. PMID- 9401433 TI - Multiple colon tumors. Diagnosis and follow-up of 450 patients with colorectal carcinoma. AB - OBJECTIVE: Failure to diagnose synchronous tumors leads to errors in patient treatment and prognosis. The existence of metachronous tumors requires strict patient follow-up to ensure early identification of the second tumor. The present study evaluates the results obtained in the application of a structured procedure for the diagnosis and follow-up of multiple colorectal carcinoma. MATERIALS AND METHODS: A structured procedure was used to follow for 5 years a group of 12 patients with multiple colorectal tumors (7 synchronous and 5 metachronous) of a series of 450 colorectal neoplasms. RESULTS: Six synchronous tumors were diagnosed preoperatively and one intraoperative. Of the 5 metachronous neoplasms, 4 strictly adhered to the follow-up protocol, as a result of which the second tumor was detected at an early stage. The remaining case involved no follow-up, and the second tumor was diagnosed in an advanced stage as a result of bowel occlusion. The left colon was predominantly involved; polyps were detected in 9 cases, while two patients had 3 malignancies detected by histopathological study. COMMENTS: We emphasize the need for a full evaluation of the colon in all patients with colorectal carcinoma. In the case of incomplete preoperative evaluation, intraoperative colonoscopy is to be considered; if this is not feasible it should be performed one month after surgery. A structured follow-up procedure permits the early detection of these tumors, there by improving patient prognosis. PMID- 9401434 TI - Hepatic tumors in patients with cirrhosis: an autopsy study. AB - The incidence and characteristics of hepatic tumors -primitive or secondary- were analyzed in a series of 596 patients with cirrhosis and on whom an autopsy was carried out. A hepatic tumor was discovered in 43.6%: 96.5% with histological findings of malignant disease and only 3.4% with benign disease. The tumors discovered showed the following in order of frequency: hepatocellular carcinoma (90.3%), hepatic metastases (4.2%), cholangiocarcinoma (2.3%), adenoma (1.5%), hemangioma (1.2%) and hamartoma (0.8%). Therefore, 10% of the neoplasms located in the cirrhotic liver were different from the hepatocellular carcinoma. In 2% of the subjects with hepatic tumors, two histologically different lesions were found to co-exist in the liver, and in every case it was found to be a hepatocellular carcinoma related to another tumor, which further complicated the diagnosis. The most frequent type of hepatocellular carcinoma was multinodular, although diffuse tumors most frequently developed metastases. When the hepatocellular carcinoma was uninodular and small, distal spread was exceptional. Metastatic infiltration of the liver by neoplasms of different origin, characteristically infrequent in cirrhosis, was always accompanied by spread to other organs and did not appear as a single nodule in any case. We conclude that the correct diagnosis of tumor related lesions located, in a cirrhotic liver is occasionally difficult during life, especially when the neoplasms are different from the hepatocellular carcinoma. PMID- 9401435 TI - New numbers to define a common plague. PMID- 9401436 TI - Nonsteroidal anti-inflammatory drug induced duodenal web. AB - Duodenal webs represent an unusual cause of intestinal obstruction in adults. These anomalies are generally considered to be congenital in origin and usually present in infancy. However, they occasionally become symptomatic in adulthood. In these cases, because of the delay in symptoms, the etiology of duodenal webs in adults is uncertain. Gastrointestinal webs in adults have also been reported in the small intestine and colon. It is generally accepted that these lesions are an acquired defect related to long term nonsteroidal anti-inflammatory drug (NSAID) use. We report a patient with a history of long term NSAID use who presented with symptoms of gastric outlet obstruction due to the presence of a duodenal web. PMID- 9401437 TI - Life would be better if we faced our fear of death. PMID- 9401438 TI - Molecular medicine: a primer for clinicians. Part X: Clinical implications of the new genetics--I. AB - The mendelian concepts of genetic inheritance remain valid and are well recognized. However, there are a number of diseases or syndromes that do not follow mendelian rules. These are designated "non-traditional patterns of inheritance" and will be the subject of the current and subsequent installments in this series. PMID- 9401439 TI - Appropriate antibiotic use in long-term care facilities. PMID- 9401440 TI - Tell your patients to stop smoking! PMID- 9401441 TI - Do the right thing for our kids. PMID- 9401442 TI - A brief history of tobacco. PMID- 9401443 TI - Tobacco update 1997: "global settlement" on hold. PMID- 9401444 TI - Tobacco update 1997: tobacco use by kids increases. PMID- 9401445 TI - Trends in lung cancer mortality among men and women, Wisconsin and the United States, 1979-1994. PMID- 9401446 TI - Smoke-free workplaces, Wisconsin municipal and county government buildings, 1996. PMID- 9401447 TI - Enforcement of minor tobacco laws: Wisconsin, 1996. AB - The objective of this project was to profile the statewide enforcement of laws prohibiting sales of tobacco to minors and purchasing/possession of tobacco by minors in 1996. A sample of Wisconsin cities or villages (n = 86) were surveyed on adoption of pertinent state statutes as local ordinances and the number of recorded violations. About 70% of municipalities have passed illegal sales ordinances and about 80% have passed purchase/possession ordinances. Over 6,000 citations were issued to minors in 1996 for purchase/possession of tobacco and 67 citations were issued for illegal sales to minors. By extrapolation, we estimate that there is approximately one commercial selling citation issued for every 112,000 packs of cigarettes sold to minors. We discuss this disparity and submit that a shift in enforcement, to commercial sellers, is needed to help reduce the increasing use of tobacco by minors. PMID- 9401448 TI - Recent advances in MR spectroscopy expand its applications in neurologic disease. AB - MRS extends the diagnostic power of MRI by displaying the biochemical composition of a selected tissue or region. When MR imaging shows a lesion, the evaluation of the chemical composition by MRS can help determine whether biopsy, observation or medical treatment is indicated. It can save some patients from biopsy prior to radiation or chemotherapy. In the future, both the image information and the spatial distribution of chemical constituents throughout the brain will be displayed with techniques such as chemical shift imaging (CSI). MRS improves the accuracy of MRI diagnosis and prognosis. MRS is performed at many sites in the country and is reimbursed by many insurers. MRS has been approved by the AMA for a CPT-4 code for reimbursement. PMID- 9401449 TI - Status epilepticus after cranial CT with contrast media. AB - This case illustrates intravenous radiographic contrast material as an unusual and little known etiology of seizure. A brief review of the literature is provided in which the incidence of contrast induced seizures and the potential risk factors for their development are discussed. PMID- 9401450 TI - What's new in ... pediatric tertiary care. PMID- 9401451 TI - Combating the fraud squad: corporate compliance programs are of utmost importance. PMID- 9401452 TI - The perception of shape and curvedness from binocular stereopsis and structure from motion. AB - The integration of binocular stereopsis and kinetic depth was measured for two distinct aspects of 3-D structure: (1) shape index, which is a measure of scale independent structure, and (2) curvedness, which is a measure of scale-dependent structure. We found that motion contributes significantly more to judged shape index than it does to judged curvedness, and stereo contributes significantly more to judged curvedness than it does to judged shape index. This suggests that the differences in the relative contribution of motion and stereo reported here occurred because these two sources do not equally specify the scale-dependent and scale-independent aspects of surface structure. Furthermore, these results seem to be inconsistent with integration models in which the different visual cues all initially contribute to the same single representation of 3-D structure. PMID- 9401453 TI - The representation of auditory source characteristics: simple geometric form. AB - Two experiments examined listeners' ability to discriminate the geometric shape of simple resonating bodies on the basis of their corresponding auditory attributes. In cross-modal matching tasks, subjects listened to recordings of pairs of metal bars (Experiment 1) or wooden bars (Experiment 2) struck in sequence and then selected a visual depiction of the bar cross sections that correctly represented their relative widths and heights from two opposing pairs presented on a computer screen. Multidimensional scaling solutions derived from matching scores for metal and wooden bars indicated that subjects' performance varied directly with increasing differences in the width/height (W/H) ratios of both sets of bars. Subsequent acoustic analyses revealed that the frequency components from torsional vibrational modes and the ratios of frequencies of transverse bending modes in the bars correlated strongly with both the bars' W/H ratios and bar coordinates in the multidimensional configurations. The results suggest that listeners can encode the auditory properties of sound sources by extracting certain invariant physical characteristics of their gross geometric properties from their acoustic behavior. PMID- 9401454 TI - Temporal binding errors are redistributed by the attentional blink. AB - When one searches for a target among nontargets appearing in rapid serial visual presentation (RSVP), one's errors in performance typically involve the misreporting of neighboring nontargets. Such illusory conjunctions or intrusion errors are distributed differently around the target, depending on task or stimulus variables. It is shown here that shifts in intrusion error patterns can be produced by the manipulation of attention alone. In a dual-task paradigm, the magnitude and distribution of intrusion errors changed systematically as a function of available attentional resources. Intrusion errors in RSVP tasks reflect internal capacity limitations for binding independent features. The present results support a two-stage model of RSVP target processing. PMID- 9401455 TI - Differential effects of stimulus context on perceived length: implications for the horizontal-vertical illusion. AB - Six experiments examined orientation-specific effects of stimulus context on the visual perception of horizontal and vertical lengths: Using a paired-comparison method, Experiments 1-5 showed that the probability of judging a given vertical line to be longer than a given horizontal line was relatively great when the stimulus set comprised relatively long horizontals and short verticals, and relatively small when the stimulus set comprised short horizontals with long verticals. To the extent that stimulus context exerts orientation-specific effects on perceived length, it thereby modulates the degree to which verticals appear longer than physically equivalent horizontals: the horizontal-vertical illusion (HVI). Under various contextual conditions, the HVI was as small as 3% (horizontals had to be 3% greater than verticals to be perceived as equally long) and as great as 15%, equaling about 12% in a "neutral" context. In Experiment 6, subjects judged the absolute physical length of each stimulus, and the results indicated that stimulus context acted largely by decreasing perceived lengths. The results are consistent with the hypothesis that differential effects of context reflect a process of stimulus-specific perceptual attenuation. PMID- 9401456 TI - A multidimensional scaling analysis of vowel discrimination in humans and monkeys. AB - Multidimensional scaling (MDS) was used to compare perceptual maps for 10 synthetic English vowels in humans and Old World monkeys (Macaca fuscata and Cercopithecus albogularis). Subjects discriminated among the vowels using a repeating background procedure, and reaction times were submitted to an MDS analysis to derive measures of perceive similarity. The dimensions that emerged related to the frequencies of the first (F1), second (F2), and third (F3) formants. Human data indicated a good match to previous MDS studies using rating procedures or confusion matrices: The dominant dimension mapped onto vowel F2, the phonetically most important formant, and the second and third dimensions mapped onto F1 and F3, respectively. For monkeys, equal weightings occurred for F1 and F2, and F3 was not clearly represented. Monkey sensitivity to the formants appeared to relate to formant amplitudes. If monkeys are giving an accurate representation of the psychoacoustic relations among the formants, then our human results suggest that species-specific mechanisms, reflecting the salience of the phonetic feature of advancement, may contribute to vowel coding in humans. PMID- 9401457 TI - Haptic curvature discrimination at several regions of the hand. AB - Static haptic discrimination of the curvature of convex, concave, or straight 20 cm-long strips was investigated for nine placements on the hand. In one condition, the strips were touched with the palmar side of the hand, and in the other condition, with the dorsal side. The influence of the lengths of the strips, and thus of contact lengths, was also investigated. For all placements, discrimination was poorer in the dorsal than in the palmar condition, owing to poorer cutaneous resolution on the dorsal side of the hand (the kinesthetic stimulation was the same in both conditions). Thus cutaneous stimulation is important. In both experiments, performance appeared to depend primarily on contact length. Moreover, the discrimination thresholds for all different placements and contact lengths followed the same trend. We conclude that in these experiments the effective stimulus for the discrimination of curved strips is the total difference of local surface attitude--that is, the slope difference over the far ends of the stimulus. PMID- 9401458 TI - Does IOR occur in discrimination tasks? Yes, it does, but later. AB - When a stimulus appears in a previously cued location several hundred milliseconds after the cue, the time required to detect that stimulus is greater than when it appears in an uncued location. This increase in detection time is known as inhibition of return (IOR). It has been suggested that IOR reflects the action of a general attentional mechanism that prevents attention from returning to previously explored loci. At the same time, the robustness of IOR has been recently disputed, given several failures to obtain the effect in tasks requiring discrimination rather than detection. In a series of eight experiments, we evaluated the differences between detection and discrimination tasks with regard to IOR. We found that IOR was consistently obtained with both tasks, although the temporal parameters required to observe IOR were different in detection and discrimination tasks. In our detection task, the effect appeared after a 400-msec delay between cue and target, and was still present after 1,300 msec. In our discrimination task, the effect appeared later and disappeared sooner. The implications of these data for theoretical accounts of IOR are discussed. PMID- 9401459 TI - The effect of shape and location on temporal masking of spatial vibrotactile patterns. AB - Target patterns presented to uncued locations on a single fingerpad were followed by either same-shape (SS) maskers or different-shape (DS) maskers presented to either the same location (SLoc) or a different location (DLoc). DS maskers interfered with identification more when they were at SLoc than when they were at DLoc, but the reverse was true for SS maskers; they interfered more at DLoc than at SLoc. When targets were presented to cued locations, performance in the absence of maskers improved, but the pattern of interference from maskers resembled that for uncued presentation. The proportion of masker responses on incorrect trials revealed that both temporal masking and response competition may be involved in the effects of location on pattern identification. PMID- 9401460 TI - Monocular discrimination of rigidly and nonrigidly moving objects. AB - We measured thresholds for the monocular discrimination of rigidly and nonrigidly moving objects defined by motion parallax. The retinal projections of rigidly moving objects are subject to certain constraints. By applying smooth 2-D transformations to the projections of rigidly moving objects, we created stimuli in which these constraints were affected. Thresholds for (generic) nonrigid transformations that in theory can be detected from rigid ones by processing pairs of views depended not only on the extent to which the rigidity constraints were affected, but also on the structure and the movement of the simulated object. Nonrigid transformations under which every three successive views had a rigid interpretation were not discriminable from rigid transformations, except in cases where the distortions were very large. Under the rigidity assumption, this would mean that a large class of nonrigidly moving objects is erroneously perceived as rigidly moving. PMID- 9401461 TI - Is visual image segmentation a bottom-up or an interactive process? AB - Visual image segmentation is the process by which the visual system groups features that are part of a single shape. Is image segmentation a bottom-up or an interactive process? In Experiments 1 and 2, we presented subjects with two overlapping shapes and asked them to determine whether two probed locations were on the same shape or on different shapes. The availability of top-down support was manipulated by presenting either upright or rotated letters. Subjects were fastest to respond when the shapes corresponded to familiar shapes--the upright letters. In Experiment 3, we used a variant of this segmentation task to rule out the possibility that subjects performed same/different judgments after segmentation and recognition of both letters. Finally, in Experiment 4, we ruled out the possibility that the advantage for upright letters was merely due to faster recognition of upright letters relative to rotated letters. The results suggested that the previous effects were not due to faster recognition of upright letters; stimulus familiarity influenced segmentation per se. The results are discussed in terms of an interactive model of visual image segmentation. PMID- 9401462 TI - The tactual horizontal-vertical illusion depends on radial motion of the entire arm. AB - We sought to clarify the causes of the tactual horizontal-vertical illusion, where vertical lines are overestimated as compared with horizontals in L and inverted-T figures. Experiment 1 did not use L or inverted-T figures, but examined continuous or bisected horizontal and vertical lines. It was expected that bisected lines would be perceived as shorter than continuous lines, as in the inverted-T figure in the horizontal-vertical illusion. Experiment 1 showed that the illusion could not be explained solely by bisection, since illusory effects were similar for continuous and bisected vertical and horizontal lines. Experiments 2 and 3 showed that the illusory effects were dependent upon stimulus size and scanning strategy. Overestimation of the vertical was minimal or absent for the smallest patterns, where it was proposed that stimuli were explored by finger movement, with flexion at the wrist. Larger stimuli induce whole-arm motions, and illusory effects were found in conditions requiring radial arm motion. The illusion was weakened or eliminated in Experiment 4 when subjects were forced to examine stimuli with finger-and-hand motion alone, that is, their elbows were kept down on the table surface, and they were prevented from making radial arm motions. Whole-arm motion damaged performance and induced perceptual error. The experiments support the hypothesis that overestimation of the vertical in the tactual horizontal-vertical illusion derives from radial scanning by the entire arm. PMID- 9401463 TI - Antiphase flicker induces depth segregation. AB - We examined the influence of the temporal phase of flickering stimuli on perceptual organization. When two regions of a uniform random-dot field are flickered in temporal alternation with the same flicker rate, one of the regions appears to lie in front of the other. Within the range of temporal frequencies used in the present experiments, depth perception was maximal between 5 and 31.3 Hz. Which region of the two is perceived as lying in front is different from person to person and sometimes fluctuates within the same subject, but when two regions are of different sizes, the smaller region tends to be perceived in front for longer than the larger region. The depth segregation was not due to a luminance difference, because the average temporal luminance of the regions was kept equal. Strikingly, the illusory depth segregation is perceived even between two adjacent regions whose densities of dots, sizes, shapes, and flicker rates are identical. This result suggests that a difference of temporal phase between two flickering regions is crucial for this new depth perception. PMID- 9401464 TI - Isoluminance and contingent color aftereffects. AB - In the typical induction of the orientation-contingent color aftereffect (CCAE), the stimuli are composed of elements that differ in both color and luminance. Three experiments are reported that show that chromatic contrast between stimulus elements is insufficient for the induction of the orientation-CCAE and that luminance contrast is necessary. These experiments expand on previous research concerned with the role of luminance contrast in the induction of orientation CCAEs by eliminating alternative explanations. PMID- 9401465 TI - Nucleotide sequence of cDNA and the gene expression of testis-specific protein Y in the Japanese monkey. AB - We cloned the cDNA for Japanese monkey Testis-Specific Protein Y (TSPY). The cDNA contained an open reading frame of 246 amino acids. This coding fragment shared 89% nucleotide sequence identity and 81% amino acid sequence identity with the homologous fragment of previously isolated human TSPY cDNA. Monkey TSPY was assumed to be a molecular mass of 28 kDa and an isoelectric point of pH 5.35. This protein was hydrophilic and contained an Arg and Lys-rich region which was a potential DNA binding site. Expression of the TSPY gene examined by reverse transcription PCR showed that the transcript was detectable only in testis, suggesting that TSPY plays an important role in spermatogenesis of primates. PMID- 9401466 TI - Bombyxin F1 gene: structure and expression of a new bombyxin family gene that forms a pair with bombyxin B10 gene. AB - Bombyxin F1 gene, a new bombyxin family gene, has been identified. The F1 gene forms a pair with bombyxin B10 gene with an opposite transcriptional orientation and the gene pair F1/B10 is located between bombyxin gene pairs B9/C1 and A7/B7 in a bombyxin gene cluster. The nucleotide sequence of the F1 gene and its deduced amino acid sequence deviate moderately from those characterized previously for the family-A, family-B, family-C, family-D, and family-E bombyxin genes; the bombyxin F1 gene and preprobombyxin F1 share no more than 62% and 53% sequence identities with other bombyxin members, respectively. Harr-plot analysis indicated that the spacer of the F1/B10 gene pair has low sequence similarity with that of other bombyxin gene pairs characterized. The bombyxin F1 mRNA in Bombyx mori brain was shown to locate in four pairs of medial neurosecretory cells, which also produce other bombyxin family mRNAs. Genomic Southern hybridization indicated that the Bombyx haploid genome contains a single copy of the family-F bombyxin gene. PMID- 9401467 TI - groE-homologous operon of Wolbachia, an intracellular symbiont of arthropods: a new approach for their phylogeny. AB - Wolbachia, a member of rickettsia found in the cells of many arthropod species, are cytoplasmically inherited bacteria which interfere with host's sexuality and reproduction. Wolbachia strains have been phylogenetically divided into A and B groups based on the nucleotide sequences of their ftsZ genes. In an attempt to further define the phylogenetical relationship among these endosymbionts, we cloned and sequenced the entire length of the groE operon of a Wolbachia harbored by a cricket. The operon encoded two heat shock proteins, which represented the third and fourth proteins of any Wolbachia ever characterized. Also, 800 bp stretches of the groE operons of several other Wolbachia were sequenced, and a phylogenetic tree was constructed based on the results. The groE tree defined the relationship among A group Wolbachia strains that had not been successfully resolved by the ftsZ tree, and suggested unexpected horizontal transmission of these bacteria. PMID- 9401468 TI - Cross purposes. Interview by Michele Bitoun Blecher. PMID- 9401469 TI - A lot is not enough. For foundations spun off by hospital sales, even billions go only so far. PMID- 9401470 TI - Founders' keepers. PMID- 9401471 TI - Is medical research doomed? AB - Managed care has stepped on the oxygen line of academic medicine, cutting patient revenue needed for research basics and delaying clinical trials. If that isn't worry enough, private firms and foreign universities are grabbing market share from academic medical centers in the United States. PMID- 9401472 TI - Pay-block time. AB - Primary care doctors pocketed huge raises five years ago, when HMOs drove up demand for their services and hospitals began snapping up their practices. But the pay party has ended, as this Executive Chartbook exclusive shows. PMID- 9401473 TI - Gold in goldenseal. AB - Healing herbs and alternative therapies already represent a market as big as $40 billion a year--with consumers paying most of the tab. That's why hospitals and entrepreneurs aren't waiting for insurers to expand coverage before venturing into alternative medicine. PMID- 9401474 TI - Contract management. Call somebody who can. AB - Specialized expertise remains the top draw for outsourcing, according to our 1997 survey of contract services. But the popularity of the most heavily penetrated services is hardly a welcome mat for expansion. In fact, it means slower growth. PMID- 9401475 TI - Contract management. Special import. PMID- 9401476 TI - Elder care. Close, but no panacea. PMID- 9401477 TI - Mental health parity. Another voice breaks the silence. PMID- 9401478 TI - Clinical guidelines. An agency for change. PMID- 9401479 TI - Managed care watch. Who's on first--patients or PR? PMID- 9401480 TI - Antibiotic resistance. Of bugs and drugs. PMID- 9401481 TI - Prenatal care. Open doors, open minds. PMID- 9401482 TI - [Structure of the natural plasminogen activator (t-PA) and of reteplase. Controversies on thrombolytic therapy. 45th yearly session of the American College of Cardiology. Orlando, Fla. March 1996]. PMID- 9401483 TI - Trends in leprosy case detection rates. AB - BACKGROUND: A systematic review of the trends in leprosy incidence is lacking. The question of whether leprosy transmission has declined remains, therefore, unanswered. This study investigates trends in new case detection rates (NCDRs) in selected leprosy-endemic areas from different continents. METHODS: A literature search using specific inclusion criteria was performed. Average annual rates of change in NCDRs were obtained by exponential curve fitting. The variation in trends within individual areas was investigated using direct and indirect information on leprosy control activities. RESULTS: This review covers 16 areas in the Pacific, Asia, Africa and Latin America. For 10 out of the 16 areas, the trend was seen to be declining consistently over the last 10 years or longer. Near stabilization or stabilization after decline was observed for two areas. For three areas, interpretation of recent NCDRs was difficult due to changes in control, but two of them showed a decline over the study period. A consistently increasing trend was observed over the last 20 years in the one remaining area. The observed downward trends could not be attributed to reduced control activities or changed diagnostic criteria. A general acceleration of downward trends in the NCDR after the introduction of multidrug therapy (MDT) has not so far occurred. CONCLUSION: Our main conclusion is that despite many differences between the studies and study areas, the review demonstrates a considerable tendency of downward NCDR trends. Lack of information and changing control conditions necessitate caution in interpreting NCDR trends in individual areas. A general impact of MDT on NCDR trends is so far not visible. The coming years will be crucial for MDT-based control to prove its ability to reduce leprosy incidence. PMID- 9401484 TI - A follow-up study of multibacillary Hansen's disease patients treated with multidrug therapy (MDT) or MDT + immunotherapy (IMT). AB - Multibacillary (MB) leprosy patients treated with multidrug therapy (MDT) or MDT + immunotherapy (IMT) with BCG + heat-killed Mycobacterium leprae were tested annually for their ability to proliferate in vitro to the mycobacterial antigens BCG, M. leprae soluble extract, and intact M. leprae. IgM antibody responses to phenolic glycolipid I (PGL-I) were measured, as well as serum nitrite levels in patients' sera, before, during and after treatment. Patients who received only MDT did not present cellular reactivity to intact M. leprae antigens, in contrast to the results obtained with BCG, which elicited reactivity at time zero, that increased after treatment. Regarding PGL-I antibody variations in relation to the initial value, we observed a statistically significant marked decrease at the end of 2 years which continued to fall in successive evaluations. MB patients showed high initial serum nitrite concentrations which dropped drastically with treatment. This decay was apparently associated with the bacillary load present in these patients. The group submitted to IMT + MDT showed high and long-lasting T-cell responses to mycobacterial antigens in a significant number of initially unresponsive MB patients. There was a marked increase to M. leprae soluble extract and BCG, as well as a more variable response to whole bacilli. The antibody levels in this group of patients are sustained for a somewhat longer period and decreased more slowly during the 5-year follow up. PMID- 9401485 TI - Progress of impairment scores following commencement of chemotherapy in multibacillary leprosy patients. AB - STUDY AIM: To investigate the progress of impairment over time in multibacillary (MB) leprosy patients. STUDY DESIGN: Retrospective cohort study. STUDY POPULATION: One-thousand-eighty-two MB patients newly registered in nine field clinics in the Western Region of Nepal between 1980 and 1993. METHODS: Data on impairment at diagnosis and at yearly intervals afterward were collected from patient records of MB patients already released from multidrug therapy (MDT). The World Health Organization (WHO) 1988 "disability" grading scale (0-2, for both eyes, hands and feet--six sites) was used as a measure of impairment. For the analysis we summed the WHO grading for the six sites into an eyes-hands-feet (EHF) sum score (minimum 0, maximum 12). The EHF score at 2 years of follow up was used to compute the main outcome measures: impairment at 2 years, yes or no, and deterioration of impairment compared with diagnosis. The combined effect of age, sex, classification and impairment status at diagnosis on the outcome was examined with logistic regression. RESULTS: At diagnosis, 55.8% of the patients had some impairment. This proportion decreased over 2 years to 43.9%. Among patients without initial impairment, 31/310 (10%) developed impairment during the study period. This was 81/396 (20.5%) among patients with impairment at diagnosis. The adjusted odds ratio (OR) for developing impairment was 1.87 [95% confidence interval (CI) 1.06-3.32] for patients with initial sensory impairment (WHO grade 1). and 1.98 (95% CI 1.15-3.4) for those with initial visible deformity (WHO grade 2). Among patients with impairment at diagnosis, 195/396 (49.2%) had improved after 2 years. CONCLUSIONS: The proportion of patients with impairment after 2 years of antileprosy treatment was 12% less than at diagnosis. Among patients without initial impairment, 10% had developed some impairment after 2 years. The risk of developing impairment was almost double for those with sensory impairment or visible deformity at diagnosis. For purposes of monitoring, evaluation and planning, both the proportion of patients with sensory impairment (WHO grade 1) and the proportion with visible deformity (WHO grade 2) should be reported at diagnosis and at release from treatment. PMID- 9401486 TI - Steroid therapy for paralytic deformities in leprosy. AB - One-hundred-forty-nine patients with 272 nerve paralyses, with visible deformity and gross disability, were prospectively followed up with steroid therapy. Out of 151 ulnar paralyses, 101 recovered (67%). Out of 52 median nerve paralyses, 45 recovered (86%); out of 69 foot drops, 54 recovered (78%) for an overall improvement of 73%. Serious side effects were few. Hence, steroid therapy should be widely encouraged for the treatment of early nerve damage to prevent permanent deformity/disability, and vigilance in spotting complications of steroid therapy is emphasized. PMID- 9401487 TI - Studies on therapeutic activity of benzoxazinorifamycin KRM-1648 in combination with other antimicrobial agents and biological response modifiers interferon gamma and granulocyte-macrophage colony-stimulating factor against M. leprae infection in athymic nude mice. AB - In the present study, we evaluated the in vivo anti-Mycobacterium leprae activities of KRM-1648 (KRM) given at long intervals in combination with ofloxacin (OFLX), clofazimine (CFZ), and dapsone (DDS). We also examined the combined effects of two biological response modifiers (BRMs), gamma interferon (IFN-gamma) and granulocyte-macrophage colony-stimulating factor (GM-CSF), on the therapeutic efficacy of KRM. KRM exhibited potent therapeutic efficacy against M. leprae infection in mice even when given at 4-week intervals. KRM displayed increased efficacy in combination with OFLX, CFZ, and DDS (given three or six times per week) when given to mice in the multidrug combination KRM + OFLX + CFZ + DDS. The therapeutic efficacy of KRM given at 4-week intervals was increased by combined use with IFN-gamma but not by GM-CSF. Adoptive transfer of M. leprae antigen-primed lymphocytes of euthymic mice to recipient athymic nude mice with progressive M. leprae infection markedly enhanced host resistance. PMID- 9401488 TI - M. leprae binds to a 28-30-kDa phosphorylated glycoprotein of rat peripheral nerve. AB - To understand Mycobacterium leprae-peripheral nerve interaction, we have investigated the binding of M. leprae to rat peripheral nerve proteins in an in vitro model using 32P-phosphorylated proteins of the peripheral nerve. Intact M. leprae binds to a major phosphorylated protein of 28-30 kDa and, to a minor extent, to a few proteins of molecular weight 45-55 kDa. This binding was more specific for M. leprae since only insignificant binding was observed with other bacteria, such as M. bovis or Escherichia coli. M. leprae did not show binding to several phosphorylated proteins of the rat brain. The 28-30-kDa binding protein of the rat peripheral nerve was found to be a glycoprotein by concanavalin A Sepharose column chromatography. PMID- 9401489 TI - A comparison of the expression of NGFr, PGP 9.5 and NSE in cutaneous lesions of patients with early leprosy using immunohistochemistry. AB - We examined the immunohistochemical expression of the neuronal proteins NGFr, PGP 9.5, and NSE in cutaneous lesions of patients with early leprosy and in the skin of normal individuals. PGP 9.5- and NSE-immunoreactive nerve fibers were decreased in the skin of leprosy patients. This reduction was topographically unrelated to the early leprosy infiltrate. However, no difference in the expression of NGFr was found between the leprosy patient and normal groups. It was shown that there is a selective alteration in the expression of neuronal proteins in early leprosy lesions which seems to be unrelated to the inflammatory infiltrate in the initial stages of leprosy. Pathogenic mechanisms other than inflammation, which are intrinsic to the Mycobacterium leprae-nerve relationship, may thus contribute to the nerve damage in leprosy neuropathy. PMID- 9401490 TI - Disseminated intravascular coagulopathy as an adverse reaction to intermittent rifampin schedule in the treatment of leprosy. PMID- 9401491 TI - Silicone oil prevention of insensitive pressure ulcers. PMID- 9401492 TI - Histoid nodules of leprosy on the lip. PMID- 9401493 TI - Minimal inhibitory concentrations of lomefloxacin and minocycline against drug sensitive and drug-resistant isolates of M. tuberculosis compared on L-J and 7H11 media. AB - The in vitro activity of lomefloxacin and minocycline was tested against 46 strains of M. tuberculosis resistant to streptomycin (S), isoniazid (H) and rifampin (R) or SHR and 51 strains sensitive to SHR by the minimal inhibitory concentration (MIC) method on two different media, namely, Lowenstein-Jensen (L J) and Middlebrook 7H11. The results of the study showed that, irrespective of the medium used, minocycline had little activity against the strains tested and the MIC was > 64 micrograms/ml. The MIC of lomefloxacin in 7H11 medium ranged from 2 to 16 micrograms/ml. There were highly significant differences in the MICs of lomefloxacin in L-J compared with 7H11. The results suggest that the activity of lomefloxacin against M. tuberculosis merits further study. PMID- 9401494 TI - The impact of molecular cytogenetics on chronic lymphoid leukaemia. AB - Chronic lymphoid leukaemias are clonal expansions of B and T cells with mature membrane phenotype. Cytogenetic study of these cases usually requires mitogenic stimulation and can often be hindered by a lack of response of the tumour cells to mitogen, poor quality metaphases, complex markers and proliferation of normal cells. In situ hybridisation with fluorescence-labelled chromosome-specific centromeric DNA probe, single or low copy sequences and whole chromosome paints which hybridise to complementary sequences allow the detection of numerical and structural abnormalities on metaphase and interphase cells with much greater efficiency. Comparative genomic hybridisation uses whole genomic tumour DNA as probe which is hybridised to normal metaphases. It is particularly useful for detecting chromosomal changes without being dependent on the dividing tumour cells. The application of these techniques to the investigation of chronic lymphoid leukaemias is reviewed with emphasis on the work done in our laboratory on trisomy 12 and the tumour suppressor region 13q14 in chronic lymphocytic leukaemia, translocation t(11;14) (q13;q32) in mantle cell lymphoma and other chronic B cell leukaemias, inv(14) (q11q32), i(8q) and complex markers in T prolymphocytic leukaemia. PMID- 9401495 TI - Levels of Hb A2 in heterozygotes and homozygotes for beta-thalassemia mutations: influence of mutations in the CACCC and ATAAA motifs of the beta-globin gene promoter. AB - The author summarizes the Hb A2 levels in over 600 beta-thalassemia heterozygotes with 32 different base changes or frameshifts and in 22 heterozygotes for 1 of 5 large deletions. Three major groups are recognized: persons with beta zero thalassemia or beta (+)-thalassemia (severe) have Hb A2 levels between 4.5 and 5.5%, those with mild beta (+)-thalassemia alleles have levels between 3.6 and 4.2%, with still lower values for those with silent mutations. High values were observed in subjects with the 2 mild beta + alleles with mutations in the beta globin gene promoter (-88, C-->T and -29, A-->G); unusually high Hb A2 values were also present in several -88 and -29 homozygotes. Data for several members of 8 families in which the -88 (C-->T) or the -29 (A-->G) mutation, or the -1,393-bp deletion, is present in cis or in trans to a delta-globin gene mutation support earlier observations that an increase in delta-chain synthesis occurs in cis to either one of these 3 alleles. A review of these data confirms the suggestion that the increase in Hb A2 levels results from at least two mechanisms: in a posttranslational system, the formation of alpha delta-dimers is promoted when excess alpha-chains are available, while certain promoter mutations increase the transcription of the delta-globin gene in cis because of a change in the binding of transcription factors. PMID- 9401496 TI - The characterisation of leukaemias with the Sysmex NE-8000. AB - Blood samples from 118 patients with acute and chronic leukaemia and with more than 50% leukaemic cells were processed with the automated haematology analyser Sysmex NE-8000. For 92 out of these 118 the differences in the histograms and scattergrams of the NE-8000 could be used in an attempt to characterise the leukaemia. This interpretation of the histograms and scattergrams appeared to be highly suggestive of the distinction between lymphatic leukaemia and myeloid leukaemia, and could be indicative of the presence of either the chronic or acute form of both the lymphatic and myeloid leukaemias. PMID- 9401497 TI - Recombinant erythropoietin trial in children with transfusion-dependent homozygous beta-thalassemia. AB - Augmentation of gamma-gene synthesis by using recombinant human erythropoietin (r Hu-EPO) represents a new approach to the therapy of beta-thalassemia. A prospective study was conducted in 26 transfusion-dependent beta-thalassemia major patients. r-Hu-EPO (Eprex/Cilag, Switzerland) was given to the patients at an initial dose of 500 IU/kg s.c. 3 times a week for at least 2 months during which no transfusion was applied. A sustained hemoglobin (Hb) level greater than 8 g/dl was considered as a response to EPO treatment. In the patients whose Hb levels remained under 8 g/dl or did not increase in comparison to pretreatment levels within 4 weeks, the dose of r-Hu-EPO was increased to 1,000 IU/kg 3 times a week and applied for another 4 weeks. Only 16 cases also received oral iron supplementation. The whole blood and reticulocyte counts, the biochemical tests including BUN, creatinine, AST, ALT, alkaline phosphatase and ferritin were done and the percentages of HbF and F cells were analyzed regularly. At the end of the 2nd month, 6 cases qualified to continue with the trial. At the end of the 6th month, r-Hu-EPO therapy was ceased in 3 cases of the 6 since their Hb levels had decreased below 7 g/dl. Only 3 cases (11.5%) continued with the r-Hu-EPO therapy without transfusion for up to 12 months. In conclusion, r-Hu-EPO may be useful in some selected transfusion-dependent patients with beta-thalassemia major. Selection criteria should include a mild beta-genotype of coinheritance of alpha thalassemia, splenectomy and pretreatment reticulocyte response of the patients as well as the patients' compliance. PMID- 9401498 TI - Recombinant human erythropoietin in the treatment of multiple myeloma-associated anemia. AB - Multiple myeloma (MM) is commonly associated with anemia. Several causes have been implicated but inadequate erythropoietin (Epo) production appears to be important. This single-institute open-label, non-comparative clinical trial was undertaken in order to evaluate serum Epo levels in patients with MM and to study the efficacy and toxicity of recombinant human Epo (rHuEpo) in the treatment of MM-associated anemia. MM patients with a baseline hemoglobin (Hb) level of < 11 g/dl received rHuEpo 150 U/kg 3 times/week subcutaneously, with a possible dose increase to 300 U/kg if no response was observed after 4 weeks. The study was designed for 12 weeks, although some responders continued rHuEpo. The study endpoints were determined by an increase in Hb and a decrease in blood transfusion requirements (BTR). Seventeen patients were enrolled in the study. The median serum Epo level was 150 mU/ml (range 11-232). Four patients did not complete the study for reasons unrelated to rHuEpo, but to their underlying MM. Twelve patients (70.6%) responded with an increase in their Hb levels. One patient (5.9%) responded partially. The median Hb level rose from 9.4 g/dl (range 7.3-10.7) at study commencement to 12.5 g/dl (range 9.0-15.2). Six of the 11 patients who were transfusion dependent enjoyed a complete abolition of BTR. The response was also interpreted as an improved quality of life: 3 patients reported a decrease of 1 level in their WHO performance status (PS) score; in 8 patients, the PS declined by 2 grades and 1 patient enjoyed PS reduction by 4 scores. Six patients continue to receive rHuEpo up to 18 months, with a good response and a smaller maintenance dose. Four patients reported flu-like symptoms, 2 suffered from a local irritation and 1 experienced a transient controlled elevation of blood pressure. SUMMARY: (1) Pretreatment endogenous serum Epo levels were relatively low in all patients studied with MM-associated anemia; (2) rHuEpo was well tolerated in these patients; (3) rHuEpo was highly effective in the treatment of anemia in MM, and (4) the response to rHuEpo is characterized by an increase in Hb levels, a reduction in BTR and an improvement in the WHO PS score. PMID- 9401499 TI - Ticlopidine-induced aplastic anemia: report of three Chinese patients and review of the literature. AB - In this study, three Chinese patients with ticlopidine-induced aplastic anemia were reported and another 13 patients in the English literature were reviewed. We attempted to find underlying similarities, evaluate the risk factors, and identify appropriate treatment for this complication. All but one of the patients were over 60 years old, and the 6 who died were all older than 65. Therefore, old age may be a risk factor for developing this complication. Agranulocytosis occurred 3-20 weeks after initiation of ticlopidine, so frequent examination of white cell count during treatment is recommended. There seemed to be no direct correlation between the dose or duration used and the severity of bone marrow suppression. Treatment for ticlopidine-induced aplastic anemia with colony stimulating factors seemed to have little effect. The fact that 5 of the 6 patients who received concurrent calcium channel blockers died, should alert clinicians to be more cautious when using these two drugs simultaneously. PMID- 9401500 TI - Thyroid tumor as initial presentation of Hodgkin's disease: a case report including an immunophenotypic characterization. AB - A rare case of Hodgkin's disease which initially presented with a thyroid tumor in an 18-year-old-man is reported. The tumor involved most of the thyroid gland but was well demarcated, and the border between the tumor and the remnants of the thyroid gland was relatively clear, suggesting secondary Hodgkin's disease involving the thyroid gland. Histologic examination of the tumor revealed nodular sclerosing Hodgkin's disease composed of many peculiar osteoclast--like giant cells and a few typical Reed--Sternberg cells in the background of dominant eosinophils. With an immunophenotypic study it was shown that these giant cells were positive for the monoclonal antibodies LeuM1 and BerH2, but negative for LCA. Thus the histological diagnosis was clearly confirmed. PMID- 9401501 TI - Acquired immune thrombocytopenia caused by IgG antiglycoprotein Ib antibody in a patient with Hodgkin's disease. AB - We studied a patient with thrombocytopenia associated with Hodgkin's disease (HD). The megakaryocyte number in the bone marrow and the level of platelet associated IgG were both increased in the patient. Intravenous gamma-globulin therapy and chemotherapy for HD dramatically normalized the platelet count, suggesting that antibody produced by lymphoma cells is likely to account for the thrombocytopenia. Antigen-captured ELISA and Western blotting showed that the patient's serum had an IgG autoantibody against platelet membrane glycoprotein Ib. The patient's plasma had no inhibitory effect on normal platelet aggregation induced by ristocetin. These findings suggest that the autoantibody found in the patient had a pathogenetic role in the thrombocytopenia, but not in platelet dysfunction. PMID- 9401502 TI - A diagnostic dilemma: chronic myelomonocytic leukemia versus atypical chronic myeloid leukemia. A case report and review of the literature. AB - There exists a great deal of overlap between many myelodysplastic syndromes and myeloproliferative disorders. This is most evident in the spectrum of disorders classified under the term chronic myeloid leukemia. These include chronic granulocytic leukemia, atypical chronic myeloid leukemia and chronic myelomonocytic leukemia. Current classification often does not clearly separate these entities since they share many features, both clinically and hematologically. We report here a case that satisfies criteria for both chronic myelomonocytic leukemia and atypical chronic myeloid leukemia, appearing to fluctuate between the two. This lends further evidence for the heterogeneity of these disorders and the need for better definition. An improved classification scheme would allow for more accurate reporting and research into etiology and treatment. The complex cytogenetic abnormalities of the case are unique and to our knowledge have not been reported previously. Also, this case report underscores the importance of cytochemical stains when such disorders are under consideration. PMID- 9401503 TI - Cytomegalovirus infection as cause of severe thrombocytopenia in a nonimmunosuppressed patient. PMID- 9401504 TI - Health risk assessment of mould allergen exposure. PMID- 9401505 TI - Dietary management of acute diarrhea with local foods in a Guatemalan rural community. AB - A community-based, randomized trial was conducted to evaluate a locally available diet for the management of acute diarrhea (n = 99 episodes) in 90 Guatemalan children, 4-42 months of age. The Test Diet (TD), a combination of a semi-solid pap (maize flour, black beans, oil) and a liquid gruel, Incaparina (maize flour, cotton seed flour, sugar), in addition to breast-milk and other home foods (group TD, n = 45 episodes) was offered for 14 d and compared to usual home feeding (group HF, n = 54 episodes). Diarrhea episodes after admission were significantly shorter for group TD (median 2.0 d) than group HF (median 4.4 d, p = 0.003) after adjusting for potential confounders. Weight gains did not differ significantly between groups. We conclude that community-based dietary management of acute childhood diarrhea using energy-dense, locally available foods is feasible and may shorten diarrhea duration. This may encourage mothers to follow recommendations for continued feeding during diarrhea in developing country environments. PMID- 9401506 TI - Failure to thrive: the earliest feature of cystic fibrosis in infants diagnosed by neonatal screening. AB - The benefits of early treatment of nutritional and respiratory problems in the CF infant and of genetic counselling for the parents are widely recognized. However, clinical diagnosis of CF is often delayed despite early onset of symptoms and the usefulness of neonatal population screening as a preventive measure is still under debate. This study analyses the clinical history of CF patients diagnosed exclusively on the basis of positive neonatal screening tests with the aim of identifying the earliest markers of the disease. We studied 103 CF infants born in north-east Italy, diagnosed following neonatal screening: assay of immunoreactive trypsin (IRT) from a heel-prick blood sample followed by a measurement of meconium lactase in cases with raised IRT. Diagnosis was confirmed by sweat test at an average age of 39 days. Eighty-one patients (79%) had symptoms strongly suggestive of CF at diagnosis, and signs and/or symptoms of pancreatic insufficiency were present in 16 of the remaining 22 cases. The most frequent symptom was growth failure (69% of infants) and of these, 44% weighed the same as at birth or less. Pancreatic insufficiency was confirmed by the low level of faecal chymotrypsin found in 85% of cases. IRT was elevated in all cases. CF had not been suspected in any symptomatic infant, although most of the infants had been monitored by a paediatrician. In conclusion, most infants with CF diagnosed by neonatal screening are already symptomatic in the first six weeks of life and the most frequent symptom is failure to thrive; pancreatic insufficiency was already present in most cases. In areas without CF neonatal screening programs, the disease should be excluded by differential diagnosis in all cases with growth failure notwithstanding adequate caloric intake in the first months of life. The high sensitivity, low cost and simple execution of IRT and fecal chymotrypsin tests make them an ideal first step in suspect cases before proceeding to the sweat test, often performed late because of limited availability. PMID- 9401507 TI - Age governs gender-dependent islet cell autoreactivity and predicts the clinical course in childhood IDDM. AB - Most IDDM patients temporarily restore some of their beta-cell function following the initiation of insulin therapy. The aim of this study was to analyse the influence of age, gender, metabolic state at diagnosis and presence of autoantibodies (GAD65 antibodies and ICA) on the duration of the clinical partial remission. In total, 149 consecutively diagnosed IDDM children, 0-16 y old (70F, 79M, mean age 9.5 y) were studied. Partial remission was arbitrarily defined as the period when the insulin dose was below 0.5 U/BW 24 h-1 and HbA1c below 7.5%, and occurred in 119/149 patients with a duration between 1 and 38 months. Cox's regression analysis showed that the factors significantly associated with the duration of remission were age, gender, interaction between age and gender, ICA and a high initial HbA1c, whereas GAD65Ab had no influence. Young boys had the shortest remission period, while adolescent boys had the longest, as compared to young and adolescent girls. The ICA-negative patients (n = 42) had a longer remission period (median 9.7 months) than the ICA-positive children (n = 107; 5.0 months; p = 0.0001), regardless of GAD65Ab status. We speculate that the relative insulin resistance, which is more pronounced in pubertal girls than in boys, may be associated with a more rapid increase of exogenous insulin requirement. These findings are important when evaluating the effect of islet cell autoreactivity on the clinical course of IDDM in children. PMID- 9401508 TI - Sub-clinical cerebral oedema does not occur regularly during treatment for diabetic ketoacidosis. AB - Fulminant cerebral oedema is an uncommon, fatal complication of diabetic ketoacidosis (DKA) in children. This study aimed to find out whether the sub clinical compression of the brain ventricles found by an earlier study, is a general phenomenon during intravenous treatment for DKA. Four boys and four girls were examined. Blood glucose values ranged from 40 to 24.6 mmol/l, base excess 34.6 to -13.6 and capillary blood pH 6.89-7.22. The patients received fluids containing both glucose and electrolytes, and insulin intravenously. After about 10h, blood glucose was 8.7-21.8 mmol/l and base excess had decreased substantially (-9.5 to -2.9) in seven of the eight cases. Computerized tomography of the brain was then performed, and again after full recovery. Only two of the patients had an initial decrease in intercaudate distance, which exceeded the variability found in a reference group. Compression of the cerebral ventricles does not occur regularly during treatment for DKA. PMID- 9401509 TI - Loperamide test: a simple and highly specific screening test for hypercortisolism in children and adolescents. AB - The overnight dexamethasone (DXM) test can give false-positive results in a few conditions (e.g. stress, strenuous exercise, depression, anorexia, anxiety, anticonvulsive therapy) in diagnosing simple obesity and hypercortisolism (HC). The loperamide (LP; a peripheral opioid agonist) test has proven useful in such conditions in adults. Thirty-one obese subjects (age 10.0-19.7 y) were studied by both overnight DXM test and LP test (8 mg orally, samples for cortisol at 0, 90, 150, 180 and 210 min) on 2 separate days. LP suppressed cortisol (< or = 138 nmol l-1) at a dose of 0.1 mg kg-1 bw (half the minimum recommended dose for the drug's antidiarrhoea effect) in 14 subjects who had normal urinary (< 4970 nmol l 1) and serum (< 552 nmol l-1) cortisol, in the absence of signs and symptoms of HC (group A). The DXM test failed to suppress cortisol in three subjects in group A, two of whom were on anticonvulsive treatment. The LP test suppressed cortisol in all of 13 subjects with elevated urinary and/or serum cortisol and/or with signs or symptoms of HC (but in whom HC was subsequently excluded on clinical grounds) (group B), while the DXM test failed to suppress cortisol in three subjects of this group. One of these was under anticonvulsive treatment and one suffered from anxiety and depression. In four patients with Cushing's syndrome (group C) neither DXM nor LP could suppress cortisol levels. Therefore, the sensitivity was 100% for both DXM and LP, while the specificity was 84% for DXM and 100% for LP. No side-effects were observed with either drug. In conclusion, LP is a useful alternative to DXM in those particular conditions that can affect its specificity in children. PMID- 9401510 TI - Moisture and mould problems in schools and respiratory manifestations in schoolchildren: clinical and skin test findings. AB - OBJECTIVE: We performed a clinical study in 99 children attending schools with moisture problems and compared the findings with those of 34 children from a reference school. The aim of the study was to evaluate the possible association between respiratory or allergic diseases in the pupils and moisture or mould problems in the school buildings. RESULTS: Asthma was diagnosed in nine (6.7%) children: eight of them came from the moisture-problem schools and all were over 10 y old. In addition, 17 non-asthmatic children had suffered from wheezing and 21 from long-term cough, both symptoms being suggestive of occult asthma. If moisture problems were observed both at home and in the school, the frequency of asthma was 21% and the combined frequency of asthma and wheezing was 43%. The presence of allergic rhinoconjuntivitis or atopic dermatitis had no association with moisture or mould problems. We performed skin-prick tests to 13 moulds in all the 133 children. A positive reaction (> 3 mm) was observed in only six (5%) of them. All six positive children reacted to at least one moisture-indicative mould, Fusarium roseum, Aspergillus fumigatus, Phoma herbarum or Rhodotorula rubra. None of these cases came from the reference school. There was a significant association between positive reactions to moisture-indicative moulds and asthma; four (44%) of the nine children with asthma had such reactions. In addition, all the 6 reactive children had either asthma or wheezing. CONCLUSIONS: We report preliminary evidence for an association between moisture or mould problems in the school building and the presence of manifest and occult asthma in the pupils. Our results show that skin-test positivity to moulds is rare in children. However, reactivity to moisture-indicative moulds seems to be associated with the occurrence of asthma or wheezing. PMID- 9401511 TI - The development of atopic sensitization in Estonian infants. AB - In a prospective study, 251 infants were followed from birth up to 12 months of age, recording manifestations of allergy by questionnaires at 3, 6, 9 and 12 months and by clinical examinations at 6 and/or 12 months. Blood samples were obtained at birth and at 6 and 12 months and analysed for serum IgE levels. The children were skin-prick tested with foods at 6 and 12 months of age and with inhalant allergens at 12 months. Blood samples from SPT-positive individuals and controls were analysed for the presence of IgE antibodies to common inhalant allergens and their cord sera for the presence of IgE antibodies to cow's milk and egg. Twelve infants (7%) were sensitized against foods [3 to cow's milk (CM) and 9 to egg white (EW)] at 6 months and 11 (5%) (2 to CM and 9 to EW) at 12 months. Seventeen infants (7%) had IgE antibodies against inhalant allergens at 6 and/or 12 months, as determined by either SPT and/or the demonstration of circulating IgE antibodies. Out of 30 children with positive SPT and/or circulating IgE antibodies against foods and inhalant allergens at any age, 6 had atopic dermatitis, 4 gastrointestinal food allergy, 1 urticaria and 4 probable allergy, while 15 had no clinical manifestation of allergy. Immunoglobulin E antibodies against Ascaris were detected in 17% of the infants with S-IgE levels > 20 kU/l. The study indicates that the incidence of sensitization and manifestations of allergic disease is similar among Estonian and Scandinavian infants during the first year of life. Given earlier findings indicating a significantly higher prevalence of atopic disease in Scandinavian school-children relative to their counterparts in Eastern Europe, the present study suggests that the key events which determine disease expression do not occur exclusively during the first year of life. PMID- 9401512 TI - Usefulness of chest physiotherapy with positive expiratory pressure (PEP)-mask in HIV-infected children with recurrent pulmonary infections. AB - Eight children with human immunodeficiency virus (HIV) and recurrent bacterial pulmonary infections were treated using a Positive Expiratory Pressure (PEP)-mask twice a day for 12 months. At the end of the study, a reduction in the number of pulmonary infections [mean (SD) 2.1 (0.9) vs 4.5 (1) p < 0.0001] and antibiotic courses [mean (SD) 1.5 (0.7) vs 2.4 (0.9) p < 0.021] was noted. The PEP-mask is a chest physiotherapy technique for removing infected secretions and optimizing airway functions that is also useful in HIV-infected children. PMID- 9401513 TI - Soluble receptors to tumour necrosis factor and interleukin-6 in urine during acute pyelonephritis. AB - We compared the urinary concentrations of soluble TNF-I (sTNF-RI), TNF-II receptors, and soluble IL-6 receptor (sIL-6R) standardized to urinary creatinine concentrations, in children with acute pyelonephritis, in children with non-renal fever and in healthy controls. These levels were related to the acute inflammatory response in the kidneys and later renal scarring, as determined by acute and 1-y follow-up with 99mTC-dimercaptosuccinic acid scintigraphy (DMSA). The concentrations of the soluble receptors were measured using enzyme immunoassay (EIA). The urinary levels of sTNF-RI were significantly higher in children with acute pyelonephritis (median 1320 pg/mmol) than in children with non-renal fever, children 6 weeks after acute pyelonephritis and healthy controls (873, 251 and 477 pg/mumol, respectively). Median sTNF-RII urine levels were also higher in acute pyelonephritis (4123 pg/mumol) than in the three control groups (2000, 964 and 1850 pg/mumol, respectively). In contrast, the highest urinary sIL 6R concentrations were found in healthy children (median 420 pg/mumol), compared to those with acute pyelonephritis (235 pg/mumol), children with non-renal fever and children 6 weeks after pyelonephritis (137 and 50 pg/mumol, respectively). No significant difference was found in any of the urinary soluble receptor levels in children with or without DMSA uptake defects at the acute or the 1-y follow-up scintigraphy. In conclusion, although the urinary soluble TNF receptor levels were higher during acute pyelonephritis, this observation was not useful for deciding which children needed follow-up after acute pyelonephritis. PMID- 9401514 TI - Cholesterol and carotid artery wall in children and adolescents with familial hypercholesterolaemia: a controlled study by ultrasound. AB - The carotid artery wall was studied with ultrasound in 23 children and adolescents with familial hypercholesterolaemia and in 23 age-matched healthy controls. The study revealed changes in the carotid artery wall related both to familial hypercholesterolaemia and to age. In the control subjects, the carotid artery wall became stiffer with age. In the patients with hypercholesterolaemia, no clear age-dependence was found, but wall stiffness correlated with total and low-density lipoprotein cholesterol. The intimal-medial wall thickness was associated with serum total cholesterol, low-density lipoprotein and triglyceride concentrations, and correlated inversely with the ratio of high-density lipoprotein to total cholesterol. Carotid artery wall properties seem to be associated with the degree of hypercholesterolaemia and the high-density lipoprotein-to-total cholesterol ratio even in children. In childhood and adolescence it is already possible, with ultrasound, to detect changes in the arterial wall related both to familial hypercholesterolaemia and to age. PMID- 9401515 TI - Sequelae of recurrent acute otitis media. Ten-year follow-up of a prospectively studied cohort of children. AB - In a cohort of 113 children prospectively followed from birth to the age of 3, two subgroups were discerned: one with recurrent acute otitis media (rAOM), the other subgroup with no AOM at all ("healthy" children). At further follow-up at the age of 10, no child had AOM or secretory otitis media (SOM), but between 3 and 7 y of age the rAOM subgroup was characterized by a significantly higher incidence of AOM as well as protracted secretory otitis media (SOM) episodes than was the "healthy" subgroup. The two subgroups did not differ significantly in hearing-thresholds at pure tone audiometry. After the age of 7, the incidence of AOM was the same in both groups. It is concluded that children with frequent AOM episodes before the age of 3 need long-term follow-up to school age, but seem not be predisposed to chronic SOM. PMID- 9401516 TI - Perinatally acquired brachial plexus palsy--a persisting challenge. AB - The aim of this investigation was to study the contemporary pattern of perinatally acquired brachial plexus palsy (BPP) in Sweden. National incidence data were collected from the Swedish Medical Birth Registry. The clinical pattern of BPP was studied in the county of Skaraborg. All children (n = 52) with confirmed neonatal BPP in 1981-89 were assessed 4-14 y after birth using routine neonatal and follow-up documentation for retrospective analysis and an assessment battery for the clinical evaluation of impairment. The mothers' recollection of the birth process was recorded by interview and compared with two control groups. The incidence of BPP in Sweden increased significantly from 1.4 per mill in 1980 to 2.3 per mill in 1994. The incidence was 45 times higher at a birthweight of > 4500 g than at a birthweight of < 3500 g. Fifty percent had a birthweight exceeding the mean +2 SD. In the Skaraborg series, half the children had normalized arm-hand function after 6 months (mean) and half had stationary impairment from 15 months (mean). Twenty-two percent of the children had severe stationary impairment of arm-hand function according to the criteria. There was no correlation between birthweight and the level of impairment. One-third of the newborn infants with BPP had neonatal care related to the difficult birth process and perinatal distress. The mothers of the children recalled the birth process as being difficult or very difficult in 77% compared with 20 and 27%, respectively, in the two control groups. This population-based investigation has revealed an unexpected increase in BPP in Sweden and has confirmed that BPP continues to be a significant cause of motor handicap in children. PMID- 9401517 TI - First-trimester invasive procedures and congenital abnormalities. AB - A prospective study was undertaken to determine whether first-trimester amniocentesis or chorion villus sampling was associated with an increased incidence of congenital anomalies. The infants of mothers who had undergone first trimester amniocentesis (EA) (n = 352), chorion villus sampling (CVS) (n = 348) or no invasive antenatal procedure (controls) (n = 264) were examined at a median age of 5 months. Both the EA and CVS groups had a higher proportion of infants with congenital anomalies (n = 18 and n = 22, respectively) than the control group (n = 4) (p < 0.01). Certain of the abnormalities, however, affected only single infants. Compression abnormalities were more common in the EA group than in the controls (p < 0.05), but not in the CVS group. The isolated limb abnormalities which occurred were minor anomalies affecting the digits and were seen in both the CVS (n = 6) and EA (n = 3) groups. First-trimester invasive procedures are thus associated with an excess of congenital anomalies. PMID- 9401518 TI - Is smoking in pregnancy interrelated between generations? AB - In order to compare pregnancy smoking habits in two generations, data on first generation deliveries in 1964-67 were obtained from prospective questionnaires. By record linkage with the Swedish Medical Birth Registry, data on daughters giving second generation births (n = 1659) were obtained, including prospective smoking information. The catchment area was that of Helsingborg County Hospital (population 145,000). Second generation births until December 31, 1993 were identified in all of Sweden. The odds ratio of maternal smoking in the first generation for pregnancy smoking among the daughters was significantly increased and approximately doubled. Adjustment for confounders was made using the Mantel Haenszel method. Similar odds ratios remained after stratification either for a social environment index of the first generation or the educational level of the second generation. Thus, our data suggest a biological association between pregnancy smoking habits over generations. Research on causative mechanisms could help design more efficient intervention programs. In a separate analysis of two parturient cohorts in Helsingborg 30 y apart, we found that the prevalence of maternal smoking had decreased comparatively more among younger than older women. PMID- 9401519 TI - Endogenous nitric oxide in the upper airways of premature and term infants. AB - Concentrations of endogenous nitric oxide (NO) were measured in premature (n = 18) and term infants (n = 7). Nasal gas was aspirated continuously and after timed occlusions, 15 s and 60 s, by a fast-response chemiluminescence analyser. The sampling flow rate was 20 ml min-1. Typical NO recordings consisted of plateaux and postocclusive peaks. In term infants peak NO concentrations (60 s occlusion) were 2.71 +/- 0.44 parts per million (ppm) within 10 min after birth, increasing (p < 0.05) to 3.81 +/- 0.25 ppm at 4-7 d postnatally. Peak NO values (15 s occlusion) averaged 1.22 +/- 0.16 ppm in premature infants (postconceptional age 25-37 weeks, body weight 623-2844 g) and the NO concentrations increased significantly with postconceptional age (p < 0.05). Nasal excretion rate, estimated from plateau NO concentrations and sampling flow rate, was 0.10 +/- 0.01 nmol min-1 kg-1 in both groups. We conclude that premature and term newborn infants excrete considerable amounts of NO in the upper airways, with hitherto not fully known functions. PMID- 9401520 TI - Maturation of hepatosomal mono-oxygenation and glucuronidation activities in pre- and full-term infants as studied using the [15N]methacetin urine test. AB - The non-distressing [15N]methacetin liver function test was modified and applied to newborn healthy infants in order to measure both the total [15N]methacetin metabolites excreted in the urine (total elimination capacity) and the proportion of glucuronated metabolite. By studying pre- and full-term normotrophic neonates 3-168 days old, the age-dependent maturation of the two developing liver function processes can be compared on the basis of either postnatal or postmenstrual age. When solely considering postnatal age, no significant differences between the pre and full-term infants were observed in the development of the total elimination capacity. However, when postmenstrual age was considered, it became apparent that this development starts earlier in pre-term infants and continues at the same rate as their full-term counterparts, up to the postmenstrual age of approximately 280 days. This increase subsequently diminishes in the pre-terms. In the same study group, the proportion of glucuronidation, another indicator of the hepatic detoxification system, appears to develop at a lower rate in pre-term than in full-term infants. When postmenstrual age is taken into consideration, glucuronidation development is also observed to begin earlier in pre-term infants and the slower maturation is more pronounced. Although these results are not generally applicable, they contribute to a better interpretation of the [15N]methacetin liver function test--for instance when estimating effects due to environmental exposure or accurately calculating age-related drug dosage for neonates. PMID- 9401521 TI - Effects of volume expansion on cardiac output in the preterm infant. AB - Clinical and echocardiographic haemodynamic evaluations of response to volume expansion are described in 12 preterm neonates aged < 7 days presenting without cardiac dysfunction and with a low cardiac output. They received 10% albumin solution (20 ml kg-1) for 3 h. Measurements were made before infusion, at volumes 5, 12.5 and 20 ml kg-1 and 1 h later. All infants increased significantly their cardiac output (CO) (from a median of 177 to 283 ml kg-1 min-1). The rise of CO decreased with the volume infused. The index of systemic vascular resistance (SVR = ratio of mean arterial pressure to the CO) decreased for the six patients without PDA (from 272 to 193 mmHg l-1 kg-1 min-1, p < 0.05) showing that the hypovolaemic preterm infant is able to shut down peripherally in response to hypovolaemia. The four hypotensive infants responded by increasing mean arterial blood pressure (from 29 to 44 mmHg). Cutaneous refilling time decreased during infusion (from 6.7 to 3.8 s. p < 0.01). One infant had an haemodynamically significant ductus arteriosus revealed by volume expansion, another one developed myocardial dysfunction. PMID- 9401522 TI - Comparison of two methods of measurement of whole blood glucose in the neonatal period. AB - The purpose of this study was to compare the performance and accuracy of the HemoCue B-Glucose photometer system and reagent strip tests used in conjunction with reflectance photometry against a reference plasma glucose method. One hundred consecutive babies admitted to the neonatal unit over a 6-month period were enrolled in the study. Each baby had a heelprick capillary glucose measured by HemoCue and reagent strip tests. At the same time venous plasma glucose and haematocrit were measured. The mean difference between the reagent strip test and plasma glucose was significantly less than the corresponding value for the HemoCue (0.015 +/- 1.41 vs 0.837 +/- 1.565 mmol l-1, mean +/- SD); however, the agreement limits between both methods and plasma glucose were wide. No significant effect of haematocrit was detected on either method. The HemoCue photometer does not offer any advantage over the widely used reagent strip tests in the neonatal period. However, the limits of agreement of both methods compared with plasma glucose are too wide to be clinically acceptable in the neonatal period. PMID- 9401523 TI - Audit of neonatal intensive care transport--closing the loop. AB - To audit the effectiveness of changes in transport arrangements, data on babies ventilated during transfer into a neonatal unit were compared between two periods. During the first period, August 1991-February 1993, an ad hoc transport team operated. Transport practice was changed in 1993 by forming a nine-person nursing transport team, improving training and upgrading monitoring. The second audit period was January 1994-July 1995. The groups were not significantly different for birthweight, gestation or levels of ventilation. Physiological variables were assessed with a "transport score". Improved scores for temperature and pH were achieved on completion of transfer in 1994-95 compared to 1991-93. Stabilizing prior to transfer took longer in the 1994-95 period. No serious deteriorations occurred in transit in the 1994-95 period, three in 1991-93. Audit facilitates identification of problems in transport. Staff, education and equipment changes were associated with improved audited outcomes. PMID- 9401524 TI - The prevalence and associated factors of epilepsy in children in Calicut District, Kerala, India. AB - To determine the prevalence of epilepsy and its association with indices of malnutrition, infection and perinatal complications in children in Calicut District, Kerala, India, a door-to-door two-stage survey was conducted in two local government districts. Among the random sample of 1172 children aged 8-12 y, 26 conformed to the definition of epilepsy giving a 5-y period prevalence of 22.2/1000. A history of perinatal complications, low BMI and recent physical symptoms were independently associated with active epilepsy. The results suggest epilepsy is highly prevalent in this population of children and that further research is needed into its cause. PMID- 9401525 TI - Oral desmopressin treatment of central diabetes insipidus in children. AB - To assess the efficacy of treatment with oral desmopressin (DDAVP), 20 patients, aged 5-20 y, with central diabetes insipidus were studied during 3 d of hospitalization and for 3 months at the outpatient clinic. At baseline the median rate of diuresis was 12.7 ml kg-1 h-1. Urinary output decreased significantly under treatment with an increase in urinary osmolality, normalization of plasma osmolality and absence of nocturia. Patients were discharged from hospital with a median dose of 500 micrograms d-1 (100-1200 micrograms d-1). An adjustment in dosage was necessary in seven patients during follow-up, resulting in a final dose of 600 micrograms d-1. Body weight and DDAVP doses (r = 0.75, p = 0.001) and body surface and DDAVP doses (r = 0.72, p < 0.001) were significantly correlated. The average dosage was 474 +/- 222 micrograms m-2 d-1 (mean +/- SD). The oral DDAVP treatment remained effective during the 3 months of follow-up. This therapy offers an alternative for the treatment of central diabetes insipidus in children. PMID- 9401526 TI - Severe skin loss after meningococcal septicaemia: complications in treatment. AB - Meningococcal septicaemia can lead to purpura fulminans with subsequent full thickness skin loss and deep muscle damage. The case reports on two infants who recovered from such a severe episode are used to describe post-septicaemic procedures and complications encountered in nursing care, psychological support and rehabilitation, with the main focus on surgery. Skin grafting is complicated by contaminated and contracting wound areas. Extensive tissue necrosis required leg amputations. Cultured keratinocytes in one of the patients were found to be too vulnerable. It has still to be proven whether more radical early-stage fasciotomies can limit skin and muscle necrosis. Patients with meningococcal septicaemia are subject to a high number of complications that are optimally treated in a burns unit. These patients require up-to-date knowledge of constantly evolving treatment possibilities and a high-level collaboration of all medical fields involved. PMID- 9401527 TI - Systemic candidiasis with acute Epstein-Barr virus infection. AB - Systemic Candida infections are usually encountered as opportunistic infections in a setting of immunologic depression. Sepsis or arthritis due to Candida is not expected in healthy people. Epstein-Barr virus may infect B cells, but does not cause immunosuppression of any clinical significance. As far as we know, invasive non-albicans Candida infection complicating Epstein-Barr virus infection has not been reported in previously healthy children. In this report, two previously healthy children, one with sepsis due to Candida species and the other sepsis and arthritis due to Candida parapsilosis are described. Both patients were male and were aged 2 and 9 y. The diagnosis was confirmed by culture. Both children also had coincidental acute Epstein-Barr virus infection, confirmed by Epstein-Barr virus viral capside antigen-IgM. They were both cured with fluconazole given for 21 days and 48 days, respectively. PMID- 9401528 TI - Multicore myopathy with restrictive cardiomyopathy. AB - A 10-y-old girl is presented who suffered mild muscular weakness and exercise intolerance from the age of 1 y onwards, with progression appearing from the age of about 8 y. Multicore myopathy and restrictive cardiomyopathy were diagnosed. Literature concerning the coexistence of multicore myopathy and cardiomyopathy is reviewed. PMID- 9401529 TI - Body surface and airway triggered ventilation in extremely premature infants. PMID- 9401530 TI - Feeding difficulties in children with cerebral palsy II. PMID- 9401531 TI - New growth hormone assays: potential benefits. AB - Three recently published new assays are described for the measurement of growth hormone (GH). Two of these--the eluted stain assay (ESTA) and the immunofunctional assay (IFA)--have been developed to measure the bioactivity of GH, rather than the immunoactivity as measured by conventional radioimmunoassays (RIAs). The third assay--the 22 kDa exclusion assay (22 k GHEA)--is designed to measure the concentrations of the different isoforms of GH present in the circulation. The ESTA is a variant of the Nb2 bioassay for lactogenic hormones, but has been adapted for specific GH measurement in serum samples. It has a lower detection limit than previous bioassays and permits the quantification of GH in large series of samples. The IFA uses a binding-site-specific antibody in combination with GH-binding protein (GHBP) in order to quantify only those GH molecules that are able to dimerize the extracellular domain of the GH receptor (GHBP), which is a prerequisite for GH signal transduction in target cells. The IFA is as convenient to use as immunoassays and can be employed routinely for GH determinations. The clinical usefulness of the 22 k GHEA has not been established, but it should provide a means of augmenting our understanding of the regulation of GH and its various isoforms. Once the ESTA bioassay or the IFA become commercially and widely available, either could replace the RIA as the standard reference method for measuring GH, as both more closely reflect the biologically active proportion of GH in serum samples than that measured by RIA. PMID- 9401532 TI - Neuropharmacological assessment of growth hormone secretion. AB - The biochemical diagnosis of individuals who are either deficient in growth hormone (GH) or who have alterations in the normal pattern of GH secretion is difficult. The uncertainty surrounding diagnosis reflects the lack of a thorough understanding of the physiology of GH secretion and of the hypothalamic hormones involved. At least three hormones are implicated: GH-releasing hormone (GHRH), somatostatin and the endogenous ligand of the GH secretagogue receptor, although the role that each plays in the release of GH is not clear from the available experimental evidence. In such a situation, most of the dynamic tests of GH secretory capacity in humans need to undergo a 'trial and error' process before being validated. The search for the 'gold standard' test of GH secretion is ongoing, and the combination of GHRH plus GH secretagogues will probably play an important role in future clinical diagnosis. PMID- 9401533 TI - An endocrinologist's approach to the growth hormone--insulin-like growth factor axis. AB - Gene knockout studies in mice, and a recent case report, have demonstrated that insulin-like growth factors (IGFs) are major mediators of pre- and postnatal growth, whereas the growth-promoting role of growth hormone (GH) appears to be confined largely to the postnatal period. The IGF axis is now known to consist of the growth factors themselves and at least seven, and probably ten, IGF-binding proteins. These act either by regulating the availability of IGFs to their receptors, or directly on their target cells. Because of the difficulties associated with GH provocative testing, the central role of IGFs in pre- and postnatal growth, and the ease of assaying the various components of the IGF axis, it is suggested that the differential diagnosis of short stature should be based on the concept of IGF deficiency rather than on GH secretory status. PMID- 9401534 TI - Secular changes in growth and maturation: an update. AB - Secular changes in growth and maturation in recent decades have been reviewed for various populations. The secular increase in attained height during the growth period is continuing in most countries, but has slowed down. The increase in adult stature over the past decades has varied between 0.3 and 3.0 cm/decade. The secular trend in the tempo of growth (earlier menarche and peak height velocity, and shortening of the growth cycle) has come to a halt in some populations, but is continuing or has been reversed in others. The secular trend in attained height and in the tempo of growth is usually more pronounced in children from low socioeconomic backgrounds, in those with poorly educated parents or in those from rural areas. It is concluded that updates of growth standards are required in all populations. More marked secular changes appear to occur in the lower height centiles, which may have direct implications on the future definition of 'short stature' in a population. PMID- 9401536 TI - The Dwarfs of Sindh: severe growth hormone (GH) deficiency caused by a mutation in the GH-releasing hormone receptor gene. AB - We report the discovery of a cluster of severe familial dwarfism in two villages in the Province of Sindh in Pakistan. Dwarfism is proportionate and occurs in members of a kindred with a high degree of consanguinity. Only the last generation is affected, with the oldest dwarf being 28 years old. The mode of inheritance is autosomal recessive. Phenotype analysis and endocrine testing revealed isolated growth hormone deficiency (GHD) as the reason for growth failure. Linkage analysis for the loci of several candidate genes yielded a high lod score for the growth hormone-releasing hormone receptor (GHRH-R) locus on chromosome 7. Amplification and sequencing of the GHRH-R gene in affected subjects demonstrated an amber nonsense mutation (GAG-->TAG; Glu50-->Stop) in exon 3. The mutation, in its homozygous form, segregated 100% with the dwarf phenotype. It predicts a truncation of the GHRH-R in its extracellular domain, which is likely to result in a severely disabled or non-existent receptor protein. Subjects who are heterozygous for the mutation show mild biochemical abnormalities in the growth hormone-releasing hormone (GHRH)--growth hormone- insulin-like growth factor axis, but have only minimal or no growth retardation. The occurrence of an offspring of two dwarfed parents indicates that the GHRH-R is not necessary for fertility in either sex. We conclude that Sindh dwarfism is caused by an inactivating mutation in the GHRH-R gene, resulting in the inability to transmit a GHRH signal and consequent severe isolated GHD. PMID- 9401535 TI - Genes regulating hypothalamic and pituitary development. AB - Several pituitary transcription factors have been identified in the last 3 years. They offer new insights into the processes that direct organogenesis, cell commitment, proliferation and differentiated function. All are DNA-binding proteins, but they have ties to different families of homeodomain proteins. They differ in their distribution and in the timing of their appearance and extinction. The Rathke's pouch homeobox protein (Rpx) has a paired-like homeodomain. In mice, it appears on embryonic day 8.5 (day e8.5) and is gone by day e14.5. Its targets for activation are unknown. Pituitary OTX has a tryptophan -phenylalanine--lysine motif in its homeodomain. It appears early and persists. It shows independent activation of the alpha-glycoprotein subunit (alpha-GSU) and pro-opiomelanocortin genes and co-operates with Pit-1 in activation of the growth hormone and prolactin genes. Pituitary Lim (P-Lim) protein also acts independently on the alpha-GSU gene, and acts in concert with Pit-1 to activate other genes. A fourth protein, termed the 'Prophet of Pit-1', or Prop-1, is the recently discovered cause of Ames dwarfism in mice. This paired-like protein is necessary for the subsequent expression of Pit-1 in somatotrophs, lactotrophs and thyrotrophs. Any or all of the newly discovered pituitary genes are candidates for mutations causing hypopituitarism in humans. As several are expressed transiently in tissues other than the pituitary during organogenesis, the phenotypes produced by mutations in these genes may prove to be complex. PMID- 9401537 TI - Insulin-like growth factor I gene deletion causing intrauterine growth retardation and severe short stature. AB - The first human case of a homozygous molecular defect in the gene encoding insulin-like growth factor I (IGF-I) is described. The patient was a 15-year-old boy from a consanguineous pedigree who presented with severe intrauterine growth failure, sensorineural deafness and mild mental retardation. Endocrine evaluation of the growth hormone (GH)--IGF-I axis revealed elevated GH secretion, undetectable serum IGF-I and normal serum IGF-binding protein-3, acid-labile subunit, and GH-binding activity. Analysis of the IGF-I gene revealed a homozygous partial IGF-I gene deletion involving exons 4 and 5, which encodes a severely truncated mature IGF-I peptide. This patient demonstrates that complete disruption of the IGF-I gene in man is compatible with life, and indicates a major role for IGF-I in human fetal growth. In addition, his neurological abnormalities suggest that IGF-I may be involved in central nervous system development. PMID- 9401538 TI - A molecular approach to sex determination in mammals. AB - Mammalian sex determination occurs in the gonad of the developing embryo. This process is dependent on the Y-chromosome-encoded Sry gene that acts in the somatic cells of the genital ridge. The transient nature of Sry gene expression suggests that it acts as a switch from one cell fate to another. One of the roles of Sry is to initiate the differentiation of Sertoli cells, which are the first cell type of the testis to be formed. Two genes are thought to be important in Sertoli cell differentiation and function, Sox9, an Sry-related gene, and SF-1, a nuclear hormone receptor. Sox9 is expressed in Sertoli cells throughout development of the mouse embryo, and inactivating mutations in this gene in humans give rise to XY females. SF-1 is also expressed in Sertoli cells and is thought to activate the AMH gene--an early marker of these cells. DAX-1, an X linked member of the nuclear hormone superfamily, is a candidate for a human condition in which duplication of regions of the X chromosome results in XY females. Expression of this gene during mouse development is associated with ovary development and is down-regulated in the differentiating testis. Mutations in DAX-1 in humans have shown that this gene is not necessary for testis development. The properties of the DAX-1 gene suggest that it is important in ovary determination and might therefore be antagonistic to the action of the Sry gene. PMID- 9401539 TI - Prader-Willi syndrome and the hypothalamus. AB - Dysfunction of various hypothalamic systems may be the basis of a number of symptoms in Prader-Willi syndrome. The often abnormal position of the baby in the uterus at the onset of labour, the high percentage of infants with asphyxia and the high proportion of children born prematurely or post-maturely may all be related to abnormal fetal hypothalamic systems, as the fetal hypothalamus plays a crucial role in labour. Abnormal luteinizing hormone-releasing hormone neurones are thought to be responsible for the decreased levels of sex hormones, resulting in non-descended testes, undersized sex organs and insufficient growth during puberty. A lack of growth hormone-releasing hormone may also contribute to the short stature of patients with Prader-Willi syndrome. In addition, the aberrant control of body temperature and daytime hypersomnolence may result from hypothalamic disturbances. The number of oxytocin neurones--the putative satiety neurones--in the hypothalamic paraventricular nucleus is markedly decreased in Prader-Willi syndrome. This is presumed to be the basis of the insatiable hunger and obesity of patients with the syndrome. PMID- 9401540 TI - The genetic basis for Prader-Willi syndrome: the importance of imprinted genes. AB - The genetic basis of Prader-Willi syndrome involves imprinted genes on the proximal long arm of chromosome 15. The basic defect appears to be the absence of function of genes that are normally expressed in a monoallelic fashion only from the paternal chromosome. In 60-70% of patients with Prader-Willi syndrome, the genetic defect is a deletion in the area of 15q11-13 on the paternal chromosome. A further 25-30% of patients with Prader-Willi syndrome do not have paternal deletions, the defect being due to uniparental disomy (UPD) for maternal chromosome 15. Paternal deletions and maternal UPD are functionally equivalent, as they both result in the absence of a paternal contribution to the genome in the 15q11-13 region. The SNRPN (small nuclear ribonucleoprotein-associated polypeptide N) gene has a critical role in the 15q11-13 region, as it is probably part of the putative imprinting centre that regulates the expression of several genes in the Prader-Willi syndrome transcriptional domain. Two further rare causes of Prader-Willi syndrome are imprinting mutations, which are microdeletions or point mutations in the putative imprinting control region, and translocations with their breakpoints in the Prader-Willi syndrome region. PMID- 9401541 TI - Hypogonadism and endocrine metabolic disorders in Prader-Willi syndrome. AB - Disturbances of the hypothalamic-pituitary-gonadal axis are reviewed in patients with Prader-Willi syndrome, and a brief account is given of thyroid function, adrenal function and glucose metabolism in such patients. Cryptorchidism, hypoplastic external genitalia and delayed or incomplete pubertal development in most patients with Prader-Willi syndrome suggest dysfunction of the hypothalamic pituitary-gonadal axis. Decreased levels of gonadotrophins, consistent with hypogonadotrophic hypogonadism, have been found in some patients, whereas others appear to have hypergonadotrophic hypogonadism secondary to cryptorchidism and its treatment. Gonadal function is normal in a small number of patients with the syndrome. Although most clinicians agree that cryptorchidism should be corrected in early childhood, in practice the surgery is often not performed. In addition, most patients do not receive sex hormone replacement therapy. It is therefore suggested that more aggressive endocrine treatment strategies for hypogonadism are warranted in both children and adults with Prader-Willi syndrome. Both thyroid function and adrenal function appear to be normal in most patients, and glucose metabolism is similar to that in normal obese individuals. PMID- 9401542 TI - Effects of growth hormone treatment on growth and body composition in Prader Willi syndrome: a preliminary report. The Swedish National Growth Hormone Advisory Group. AB - A controlled, randomized study was conducted to assess the effect of growth hormone (GH) treatment on growth, body composition and behaviour in prepubertal children (3-12 years of age) with Prader-Willi syndrome. GH treatment was given to one group of 15 patients (group A) at a dose of 0.1 IU/kg/day for 2 years. The second group (group B; n = 12) was not treated for the first year and was then given GH at a dose of 0.2 IU/kg/day for the second year. All patients had low 24 hour levels of GH and insulin-like growth factor I before GH treatment. Height velocity SDS increased from -1.9 +/- 2.0 to 6.0 +/- 3.2 during the first year of GH treatment in group A, and from -1.4 +/- 1.2 to 10.1 +/- 3.9 in the second year of the study in group B. When GH treatment was stopped, height velocity declined dramatically. Height SDS followed a similar pattern. GH treatment reduced the percentage body fat and increased the muscle area of the thigh. Isometric muscle strength was also increased. In addition, GH treatment appeared to have psychological and behavioural benefits, which were reversed after cessation of treatment. It was concluded that GH treatment improves growth, body composition and behaviour in children with Prader-Willi syndrome. PMID- 9401543 TI - Effect of 6 months of growth hormone treatment in young children with Prader Willi syndrome. AB - Nine prepubertal children with Prader-Willi syndrome were treated with growth hormone (GH; 24 IU/m2/week) for 6 months. Mean height increased by 0.8 SD and mean weight for height decreased by 0.7 SD over this 6-month treatment period. Body fat, measured by dual-energy X-ray absorptiometry, decreased by 22.5% over the period of GH treatment, whereas fat-free mass increased by 14%. These preliminary results indicate that GH is effective in increasing height and normalizing body composition in patients with Prader-Willi syndrome. PMID- 9401544 TI - Growth hormone, insulin-like growth factor and the immune system. PMID- 9401546 TI - Effects of growth hormone and insulin-like growth factor I on T- and B lymphocytes and immune function. AB - The concept of a neuro-endocrine-immune axis was proposed more than 50 years ago. Growth hormone (GH), a central component of this axis has many functions at both a molecular and cellular level, including thymocyte proliferation, stimulation of the cytotoxic activity of natural killer cells and induction of lymphocyte proliferation. Binding of GH to its receptors on lymphocytes stimulates the production of insulin-like growth factor I (IGF-I), which mediates the effects of GH on cell proliferation. Other effects of GH on the immune system appear to be direct, such as priming monocytes for enhanced production of hydrogen peroxide in response to phorbol esters, and stimulating neutrophils to secrete superoxide anions associated with enhanced phagocytic activity. Many of the effects of GH are shared by IGF-I. Despite these observations, and the fact that GH is produced and secreted in immunological tissues such as the thymus and spleen, immune deficiency is not characteristic of GH deficiency in humans. The question remains as to whether GH and IGF-I could be used as immunotherapy. Currently, both agents have been used in adults to diminish wasting due to acquired immunodeficiency syndrome, and GH has been shown to stimulate CD8+ cell counts. However, they had little impact on CD4+ cell counts, which may be due to IGF-I and GH resistance in these individuals. The use of GH and IGF-I as immunotherapies merits further study. PMID- 9401545 TI - Pituitary hormones and immune function. AB - The pituitary gland plays a key role in the regulation of growth, differentiation and function of all cells in the body, including immunocytes. Immune reactions are generated through the proliferation of antigen-specific lymphocyte clones. Growth hormone and prolactin are required for the development of mature lymphocytes and for the maintenance of immunocompetence. These hormones enable lymphocytes to respond to antigen, which is delivered as an adherence signal in the context of major histocompatibility surface molecules of antigen-presenting cells. Numerous other adhesion molecules play a role in the regulation of lymphocyte activation. The activation process is completed by cytokine signalling, after which lymphocyte proliferation, differentiation and functional maturation take place. Interleukins, hormones and growth factors may all function as cytokines. Many lymphocytes exist in the body in a quiescent state, with minimal metabolic activities. These cells are maintained by competence hormones and insulin-like growth factor 1, which are present in the systemic and local environment. Apparently, some steroid hormones, opioid peptides and catecholamines are capable of modulating delivery of the signal from the lymphocyte membrane receptor to the nucleus. Steroid and thyroid hormones control nuclear transcription factors as their receptors, and thus are powerful regulators of lymphocyte signalling at the nuclear level. The bioactive forms of thyroid hormone and of several steroid hormones are generated locally by immunocytes. These important hormonal immunoregulators function both at systemic and local levels. Glucocorticoids are major regulators of cytokine production, and alpha-melanocyte-stimulating hormone functions as a powerful cytokine antagonist. The hormones secreted or regulated by the pituitary gland therefore regulate every level of immune activity, including the competence of lymphocytes to respond to immune/inflammatory stimuli, signal transduction, gene activation, the production and activity of cytokines and other immune effector functions. PMID- 9401547 TI - Effects of growth hormone and insulin-like growth factor I binding to natural killer cells. AB - Human growth hormone (GH) and insulin-like growth factor I (IGF-I) are known to bind to, and exert modulatory effects on, different immunocompetent cells, including CD16+/CD3- natural killer (NK) cells. NK cells are involved in various non-major-histocompatibility-complex-restricted actions of the immune system. Although no clinically significant defect in tumour or virus defence has been reported in GH-deficient patients, the data available indicate decreased NK cell activity in these patients. In most studies, the absolute number and percentage of NK cells have been found to be normal. Substitution with GH has been reported to normalize the decreased NK cell activity in GH-deficient patients. In a cross sectional study in GH-deficient adults, decreased basal and interferon-beta (IFN beta)-stimulated NK cell activity in vitro. Preliminary data on GH binding to NK cells indicate enhanced binding in GH-deficient patients when compared with normal controls. PMID- 9401548 TI - Role of B-cells in growth hormone-immune interactions. AB - Cells demonstrating cell surface markers for B-cells tend to be normal in growth hormone (GH)-deficient children. Treatment with pituitary-derived or recombinant human GH, however, produces a significant, albeit transient, decrease in the number of these cells both in vitro and in vivo. Receptors for GH have been detected on numerous cells of the immune system, notably B-cells and monocytes. Recent studies have shown that cells of the immune system are able to produce peptide hormones, such as GH, and studies in the rat (which have yet to be confirmed in humans) suggest that the B-cell is the cell type most involved in this production. These findings suggest that the B-cell has a significant role in the bi-directional communication network between the endocrine and immune systems. PMID- 9401549 TI - Effects of growth hormone on natural killer cells. PMID- 9401550 TI - The development of 'impervious peptides' as growth hormone secretagogues. AB - The discovery and development of growth hormone (GH) secretagogues is briefly reviewed. GH-releasing peptide-6 (GHRP-6) was the first GHRP to be developed that was active in vivo. Smaller peptides were found to have no GH-releasing activity in vivo, even though they were potent releasers of GH in vitro. Substituting Trp with 2-Me-Trp led to the development of orally active hexarelin and other, smaller, peptides. Although they showed oral activity, absorption rarely exceeded 1% of the administered dose. Nonpeptide GH secretagogues have now been developed that combine reasonable oral absorption with high potency. The next development will be to produce GH secretagogues that overcome the lack of specificity shown by the present molecules. PMID- 9401551 TI - Hypothalamic targets for growth hormone secretagogues. AB - Various novel growth hormone (GH) secretagogues have been developed. GH secretagogues release GH directly from the pituitary via a pathway distinct from that involving GH-releasing hormone (GHRH). However, they also act centrally to activate hypothalamic neurones, and require an intact GHRH system for potent in vivo activity. Both normal and transgenic growth-retarded (Tgr) rats release GH in response to GH secretagogues, and their responses are sensitive to the pattern of secretagogue administration. GH secretagogues are not completely specific for GH release, but also activate the adrenocorticotrophin-adrenal axis, implying that they have additional central actions. The recent cloning of an endogenous receptor for GH secretagogues now makes it possible to identify central targets for their action. An endogenous receptor implies the existence of an endogenous ligand, but its site of production, relationship to the xenobiotic pharmacological agents and its underlying physiological relevance remain unclear. PMID- 9401552 TI - Age-related growth hormone-releasing activity of growth hormone secretagogues in humans. AB - Growth hormone-releasing peptides (GHRPs) are synthetic molecules with strong, dose-related and reproducible growth hormone (GH)-releasing activity in humans. GHRPs act at both the pituitary and the hypothalamic level, where specific receptors have been located. In adults, GHRPs release more GH than does GH releasing hormone (GHRP), whilst their co-administration has a synergistic effect, indicating that they have, at least partially, different mechanisms of action. However, normal activity of GHRH-secreting neurones is needed to achieve the full GH-releasing effect of GHRPs. In contrast to GHRH, the GH-releasing activity of GHRPs is not further increased by substances acting via inhibition of hypothalamic somatostatin, and is only blunted by substances that stimulate hypothalamic somatostatin release. Even free fatty acids and exogenous somatostatin, which act directly on somatotrophs, do no more than blunt the effect of GHRPs. Thus, the GH-releasing activity of GHRPs is partially refractory to inhibitory influences, GHRPs act, at least in part, by antagonism of somatostatin activity, both at the pituitary and the hypothalamic level. The GH releasing effect of GHRPs is not dependent on gender, but undergoes age-related variations. Gonadal steroids seem to influence the activity of GHRPs only in childhood. The reduced GH response to GHRPs in the elderly is probably due mainly to concomitant GHRH hypoactivity and somatostatinergic hyperactivity. A preserved GH-releasing effect of GHRPs has been reported in acromegaly, anorexia nervosa, hyperthyroidism and in critically ill patients. GHRPs have also been found to increase GH release in children with idiopathic short stature, in GH deficiency and in obese patients, in whom there is a well-known reduction of somatotroph function. The GH response to GHRPs is markedly reduced in hypothyroidism and Cushing's syndrome. PMID- 9401553 TI - Growth hormone secretagogues in pathological states: diagnostic implications. AB - The identification and cloning of the receptor for synthetic growth hormone (GH) secretagogues, even before the endogenous ligand has been identified or its precise physiological role established, suggests that there is a novel target of action for this class of drug. In an attempt to select patients who will benefit from GH treatment, GH secretagogues are being evaluated for their usefulness in diagnosing GH deficiency. The effects of GH-releasing peptides (GHRPs) on GH release as a function of age and metabolic status, and in different neuroendocrine pathologies, are described, as are the different mechanisms of action, potency and reproducibility of the response to GHRPs compared with GH releasing hormone (GHRH). GHRPs offer the advantage over GHRH in natural models of deranged GH secretion in that, in various metabolic states (e.g. obesity, anorexia nervosa and non-insulin-dependent diabetes mellitus), the GH response to GHRH is more impaired than it is to GHRPs. However, in some neuroendocrine pathologies, the reverse is true. Thus, both secretagogues provide separate information on the physiological status of somatotrophs. PMID- 9401554 TI - Growth hormone secretagogues in children with altered growth. AB - A diagnostic test was devised to evaluate pituitary growth hormone (GH) secretory potential. GH secretory dynamics were assessed in children with and without GH deficiency. The GH response was measured to GH-releasing hormone (GHRH) and the GH-releasing peptide GHRP-2, administered sequentially. The mean (+/- SEM) peak GH response to GHRP-2 was 20.1 +/- 5.5, 63.6 +/- 24.9 and 42.2 +/- 4.3 micrograms/l for GH-deficient, slowly growing non-GH-deficient and control children, respectively (p < 0.02 and p < 0.05 for GH-deficient vs controls and slowly growing children, respectively). Corresponding values for area under the curve (AUC) were 995 +/- 371. 2460 +/- 953 and 1598 +/- 274 micrograms/l x minute. Peak GH (and AUC) responses to GHRH were 19.6 +/- 5.1 micrograms/l (924 +/- 232 micrograms/l x minute), 31.4 +/- 8.4 micrograms/l (1544 +/- 449 micrograms/l x minute) and 39.8 +/- 7.8 micrograms/l (2201 +/- 437 micrograms/l x minute) for the same three groups, respectively (p < 0.05 for peak GH in GH deficient patients vs controls, and p < 0.02 and p < 0.01 for AUC in GH-deficient vs slowly growing children and controls, respectively). The ratio of the peak GH response to GHRP-2 and GHRH was similar in all three groups. As these secretagogues stimulate different aspects of hypothalamic function (i.e., they are functional complements), robust GH secretion in response to GHRH or GHRP could suggest adequate endogenous GHRP or GHRH, respectively. A poor response to either GH secretagogue administered individually could represent inadequacy of its endogenous complement. The integrity of functional pituitary elements could be differentiated from inadequate complements by administering both GH secretagogues simultaneously. Application of these principles should allow a better definition of the underlying disorder and provide the basis for therapeutic strategies for those patients with abnormal GH production and/or secretion. PMID- 9401555 TI - The biology of bone maturation and ageing. AB - The only indicator of development that is available from birth to maturity is skeletal age. This short review discusses how standard bone ages have been developed from assessment of radiographs, and describes the advantages and disadvantages of the 'atlas' approach as developed by Greulich and Pyle, and the bone by bone approach, as developed by Tanner. As the standards currently available are based mainly on historical series of radiographs from particular populations, it is stressed that national standards should be established and updated regularly if bone ages are to be used to assess development. The question of the clinical relevance of using bone age assessments of the hand and wrist to determine the state maturation of the whole skeleton and particularly, the growth potential is also discussed. It is concluded that, despite the difficulties of assessing bone age, and the assumptions on which the various methods are based, determination of skeletal development is clinically relevant in that it provides the only means of assessing rates of maturational change throughout the growing period. PMID- 9401556 TI - Variation of bone age progression in healthy children. AB - Bone age assessments were related to auxological variables in 407 Italian boys, between 7 and 12 years of age, in order to elucidate the factors that affect the rate of skeletal maturation and to examine the possibility of using measures of skeletal maturation of evaluate individual patients. Using the radius-ulna-short bones (RUS) method of assessment, bone age velocity was greater in the Italian boys than for the UK reference standards, although there was considerable interindividual dispersion around the mean. Bone age velocity and height velocity were poorly correlated, and there was little correlation between skeletal and pubertal maturation. There was a slight positive correlation between bone age velocity and height SDS and between bone age velocity and body mass index. Bone age estimations using RUS were greater than those obtained using the carpus. In conclusion, the marked interindividual deviation in measured bone ages makes it difficult to relate data on an individual basis to other measures of growth and maturation PMID- 9401557 TI - Population-specific reference values for bone age. AB - Contemporary reference values for assessing skeletal maturity have been obtained for the Japanese population. These were used to compare skeletal maturation with populations from the UK. Belgium, North India and South China. Japanese children were found to attain skeletal maturity, based on measurements of the radius, ulna and short bones of the left hand and wrist, 1 or 2 years earlier than present-day European and Chinese children. A relative lack of data for the North Indian population made comparison impossible. PMID- 9401558 TI - Clinical usefulness of bone age determination in the management of tall stature. AB - Measurements of skeletal maturation are used in the study and management of growth and growth disorders because of the correlation between the degree of skeletal maturation and the potential for further growth. In a study of 147 tall girls, final height was overpredicted by 0.7 +/- 2.5 cm (mean +/- SD) when based on the Bayley-Pinneau method. The radius-ulna-short bones (Tanner-Whitehouse 2) mark II method of assessment produced an underprediction of final height by 0.8 +/- 2.9 cm and 1.0 +/- 2.8 cm when the rating was performed by a skilled human or by a computer-aided system, respectively. These errors in prediction were considered acceptable, although when predictions were made on five girls below 11 years of age, they were less accurate. PMID- 9401559 TI - Diagnosis of severe growth hormone (GH) deficiency in young adults who received GH replacement therapy during childhood. AB - Between 20% and 87% of young adults who had completed growth hormone (GH) therapy in childhood for a putative diagnosis of GH deficiency (GHD) had normal GH responses to provocative tests when they were retested. Patients with isolated idiopathic GHD were more likely to exhibit normal GH responses at retest in young adult life than were patients with multiple pituitary hormone deficits. When determining which patients should receive GH therapy in adult life, those who have isolated GHD should undergo two tests of GH status, while those with multiple anterior pituitary hormone deficits require only one test. Most information is available for the insulin tolerance test, the arginine stimulation test and the glucagon stimulation test, but more recent methods, such as GH releasing hormone in combination with pyridostigmine, are showing promise in the investigation of GHD. In young adults with childhood-onset GHD, the serum concentration of insulin-like growth factor I is a useful marker of GH status, and can be used in conjunction with a GH provocative test. The choice of GH provocative test should ultimately depend on the experience and policy developed at the centre performing the assessment. Whichever tests are chosen, each should be validated in subjects known to have hypothalamic-pituitary disease as well as in normal individuals. PMID- 9401560 TI - Can we predict and prevent adult morbidity in males with childhood-onset growth hormone deficiency? AB - In order to achieve an optimal quality of life and minimize morbidity in adulthood, the therapeutic management of patients with childhood-onset growth hormone deficiency (GHD) should follow strict guidelines. Optimal final height should be obtained by the early diagnosis of GHD and subsequent adequate growth hormone (GH) dosing and duration of treatment. Moreover, particularly in males with associated gonadotrophin deficiency, the psychosexual maladjustment could be prevented by the earlier induction of puberty, completion of the male phenotype and testicular stimulation with gonadotrophins or luteinizing hormone-releasing hormone. Finally, a satisfactory peak bone mass could be attained by continuation of GH treatment for some years after cessation of linear growth. PMID- 9401561 TI - Conditions spanning paediatric and adult endocrine practice--the adult perspective. AB - Close liaison between paediatric and adult endocrinologists is essential for optimum care in a variety of clinical conditions. The increasingly recognized importance of growth hormone deficiency (GHD) in the adult is a further indication for maintaining long-term follow-up of patients with isolated GHD, which remains demonstrable when linear growth is complete, in addition to those patients presenting in childhood with evidence of structural pituitary disease and anterior pituitary failure. Additional areas in which liaison is desirable include congenital adrenal hyperplasia, precocious puberty, gonadal dysgenesis and other disorders of primary and secondary sexual development, thyroid dysfunction, diabetes mellitus, inherited neoplasia syndromes and those conditions, for example Cushing's syndrome, which present in childhood but are more common in adult clinical endocrine practice. In this brief review, the diagnostic spectrum of the paediatric/adult interface is described and the rationale for an integrated approach to treatment and follow-up is outlined. PMID- 9401562 TI - When and how to transfer patients from paediatric to adult endocrinologists: experience from St Bartholomew's Hospital, London. PMID- 9401563 TI - Endocrine and nutritional regulation of prenatal growth. AB - Fetal growth in late gestation is primarily determined by the functional status of the pathways by which nutrients are transferred from the maternal compartment, across the placenta, and taken up by the fetal tissues. Both the maternal and fetal endocrine systems can influence this pathway at several levels, including regulation of placental metabolism. The primary fetal axis involved in the regulation of fetal growth is the glucose-insulin-insulin-like growth factor I axis. PMID- 9401564 TI - Early influences on embryonic and placental growth. AB - Growth of the placenta is influenced by events before and during early pregnancy. Some of these events set the growth trajectory of the placenta and the fetus for the remainder of the pregnancy. Maternal size and nutrition, and the local metabolic, cytokine and hormonal environment of the embryo all affect growth of the placenta. PMID- 9401565 TI - Morphometry of fetal growth. AB - Intrauterine growth retardation (IUGR) complicates about 5% of all pregnancies and is responsible for substantial perinatal mortality and morbidity. With ultrasound, it is possible to assess fetal brain growth indirectly by measurement of the biparietal diameter, head circumference and transcerebellar diameter. As liver size is affected most profoundly by IUGR, measurement of the abdominal circumference of the fetus provides the earliest evidence of fetal growth restriction. Placental size, assessed with ultrasound, can also indicate the severity of the condition. Once a fetus is diagnosed as having IUGR, its well being can be monitored with standard heart-rate testing and ultrasound assessment of fetal behaviour. Doppler analysis of the wave form of the fetal arterial and venous circulation hold the greatest promise for managing pregnancies complicated by IUGR. PMID- 9401566 TI - Pathophysiology of intrauterine growth retardation: role of the placenta. AB - The placenta is essential for normal fetal development. Failure of the placenta can result in many fetal conditions, for example, intrauterine growth retardation (IUGR). Placentas from pregnancies complicated by IUGR show vascular damage, which may lead to the onset of pregnancy-induced maternal hypertension. Accurate placental assessment may, therefore, indicate which fetuses are at risk of IUGR and so improve clinical evaluation and management of both the fetus and the mother. Placental development and function can be assessed by a number of methods, including measurement of placental weight at mid-gestation (placental growth in the second trimester correlates strongly with placental weight at birth), assessment of fetal and placental circulation (an association between perinatal morbidity and abnormal blood velocity profiles has been established) and assessment of placental metabolism and nutritional transfer (a reduction in transfer of nutrients may be an early indicator of IUGR. PMID- 9401567 TI - Aetiology, morphology and body composition of infants born small for gestational age. AB - Infants born small for gestational age (SGA) are a heterogeneous group. Both the timing and duration of the intrauterine insult determine the physical condition and body composition of the infant at birth. Infants with symmetrical intrauterine growth retardation (IUGR) have a similar body composition at birth to weight-matched infants born appropriate for gestational age. However, these infants are more likely to remain shorter and lighter than normal infants. In contrast, infants with asymmetrical IUGR have reduced fat deposition but are more likely to exhibit catch-up growth during the first few months of life. The low mortality and morbidity rates in infants born SGA observed in recent studies are linked to their appropriate perinatal management, including adequate early nutritional support. PMID- 9401568 TI - Intrauterine programming of coronary heart disease and stroke. AB - We have become accustomed to the idea that the major disorders of adult life, including coronary heart disease, stroke and diabetes, arise from an interaction between influences in our adult lifestyle and a genetically determined susceptibility. Recent research, however, suggests that growth in utero may also play an important role. PMID- 9401569 TI - Metabolic consequences of intrauterine growth retardation. AB - Epidemiological studies have revealed strong and reproducible links between indices of poor fetal, and possibly infant, growth and susceptibility to the development of glucose intolerance and insulin resistance syndrome in adult life. The 'thrifty phenotype' hypothesis has been proposed to explain these associations. Key features of the hypothesis are: (i) intrauterine growth retardation has a nutritional basis and the resulting altered fetal environment permanently alters the development and metabolic functions of organs: (ii) these alterations are beneficial to survival in a poor nutritional environment, but may lead to diseases such as non-insulin-dependent diabetes mellitus if nutrition is abundant and obesity occurs in adult life. Tests of this hypothesis in an animal model in which pregnant and/or lactating rats were fed a diet with a reduced protein content have shown that liver metabolism in the offspring is permanently altered despite their being weaned onto a normal diet. The longevity of male offspring may be significantly increased or decreased depending on whether growth retardation is restricted to the period of suckling or pregnancy, respectively. The latter finding raises questions about potentially detrimental effects of 'catch-up' growth. PMID- 9401570 TI - Hormonal status of short children born small for gestational age. AB - The present study was undertaken to evaluate the hormonal status in a subgroup of prepubertal children born small for gestational age (SGA) who lacked postnatal catch-up growth. In this subgroup, a reduced rate of growth hormone (GH) secretion was found, compared with reference groups of healthy children born appropriate for gestational age, of either normal or short stature at the time of investigation. In addition, an abnormal pattern of GH secretion was observed in short children born SGA, which was most pronounced in the younger children, and involved an increased frequency of GH peaks of low amplitude, combined with increased baseline secretion. Levels of insulin-like growth factor I (IGF-I) and IGF-binding protein-3 were also reduced in short children born SGA, compared with the reference groups. These findings may explain, in part, the lack of postnatal catch-up growth in short children born SGA. PMID- 9401571 TI - Postnatal growth of children born small for gestational age. AB - A large, population-based representative study (n = 3656) has shown that the vast majority of healthy, full-term, singleton infants born small for gestational age (SGA) achieve catch-up growth during the first 2 years of life. Indeed, most of the increase in height SDS occurs by 2 months of age. Children born SGA who do not show postnatal catch-up growth and so remain short at 2 years of age, have a higher risk of short stature (< -2 SDS) in later life, with a relative risk at 18 years of age of 5.2 if born light and of 7.1 if born short. PMID- 9401572 TI - Antenatal therapy for intrauterine growth retardation. AB - Currently, there is no effective antenatal therapy for intrauterine growth retardation (IUGR). Although the IUGR fetus is undernourished in utero and there have been many attempts to treat IUGR with nutritional supplements, most studies have been poorly controlled, and there is no evidence to date that nutrient supplements can reverse the process of IUGR once it is established. Nutrient supplementation is also potentially risky and a combination of nutrients is likely to be needed. Alternative approaches to antenatal therapy for IUGR that show promise include fetal growth hormone and insulin-like growth factor I treatment to improve fetal growth. Fetal and maternal hormone supplements may also prove useful in IUGR by improving placental function and thus fetal substrate supply. Fetal enteral supplementation by the administration of growth factors and/or nutrients into the amniotic fluid may also prove effective and clinically feasible. It seems likely that combinations of these approaches will be required before effective therapy can be devised for the IUGR fetus in utero. PMID- 9401573 TI - Nutritional management and growth hormone treatment of preterm infants born small for gestational age. AB - In severe cases of intrauterine growth retardation, elective preterm delivery may provide the possibility for nutritional intervention to prevent some of the long term consequences of the catabolic condition in utero. Neonatal nutritional management is aimed at providing a high protein intake of up to 4 g/kg/day in order to obtain the rapid increase in protein that is seen in normally growing infants during the early postnatal period. Unfortunately, due to impaired production of urea, high plasma levels of ammonia, which may be rate limiting with respect to an optimal gain in protein, are often observed in the preterm infant born small for gestational age (SGA). In an attempt to stimulate protein synthesis in preterm infants born SGA, growth hormone (GH) treatment was given to seven such infants during the early postnatal period. The infants received daily subcutaneous injections of GH (1.0 IU/kg/day) from postnatal day 7 until a body weight of 2000 g was reached (postnatal week 7-8). A further seven preterm infants born SGA were studied as controls. GH treatment had no significant effects on growth, body composition, net protein gain and glucose metabolism. Furthermore, plasma levels of insulin-like growth factor I (IGF-1) and IGF binding protein-3 revealed a normal developmental increase and were not significantly altered by GH treatment. These results may be explained by a relative GH insensitivity or resistance during this period of early preterm life. PMID- 9401574 TI - Growth hormone treatment of short children born small for gestational age: reappraisal of the rate of bone maturation over 2 years and metanalysis of height gain over 4 years. AB - A minority of children born small for gestational age (SGA) fail to achieve sufficient catch-up growth during infancy and remain short throughout childhood, apparently without being growth hormone (GH) deficient. A previous metanalysis of four trials revealed that GH treatment over a period of 2 years induced a dose dependent acceleration of linear growth and, to a lesser extent, of the rate of bone maturation in short, prepubertal children born SGA. The rate of bone maturation and the change in height SDS for bone age from the previous 2-year metanalysis have been re-analysed according to chronological age (two prepubertal age groups: group A, 3.0-5.9 years old; group B, 6.0-8.9 years old). The rate of bone maturation was slower in younger than in older prepubertal children; this difference was more marked in children receiving high-dose (0.2 or 0.3 IU/kg/day) GH treatment (p < or = 0.01). Accordingly, the change in height SDS for bone age was increased by high-dose GH treatment in both age groups (p < or = 0.01), and was more pronounced in younger than in older children (1.45 +/- 0.28 versus 0.63 +/- 0.20; p < or = 0.01). Height SDS data from 100 short, prepubertal children born SGA have been analysed over 4 years. The change in height SDS appeared to be related to the average dose of GH. A mean GH dose of 0.1 IU/kg/day over 4 years was administered either as 0.1 IU/kg/day for 4 years (continuous) or as 0.2 IU/kg/day for 2 years, followed by 2 years without GH treatment (discontinuous). After 4 years of treatment, the increase in height SDS for the continuous and discontinuous treatment schedules was similar, being 1.42 +/- 0.10 SDS and 1.58 +/- 0.17 SDS, respectively. In a second regimen, a mean GH dose of 0.2 IU/kg/day over 3 years was administered either as 0.2 IU/kg/day for 3 years (continuous) or as 0.3 IU/kg/day for 2 years, followed by 1 year without GH treatment (discontinuous). After 3 years, the increase in height SDS with the continuous and discontinuous treatment schedules was similar, being 2.01 +/- 0.18 SDS and 2.22 +/- 0.16 SDS, respectively. GH administration was well tolerated in all treatment groups. In conclusion, the rate of bone maturation in short, prepubertal children born SGA treated with GH appeared to depend not only on the dose of GH, but also on the age of the child. GH treatment resulted in a prolonged increase in height SDS, the magnitude of the rise being dependent on the average GH dose rather than on the continuous or discontinuous mode of GH administration. PMID- 9401575 TI - Growth hormone treatment of short children born small for gestational age. AB - The administration of growth hormone (GH) for a short period to short children born small for gestational age increases growth velocity. Children receiving 2-3 times the replacement dose gained nearly 2.0 SDS in height over a 3-year treatment period. This treatment is well tolerated, without significant side effects. Further studies of the growth rate following cessation of GH treatment will reveal whether the gain in height is maintained, and whether the final height prediction is improved. PMID- 9401576 TI - Intrauterine growth retardation: our current understanding and future directions. PMID- 9401577 TI - Can reactive oxygen species precondition the isolated rat heart against arrhythmias and stunning? AB - Ischaemic preconditioning has cardioprotective effects. Reactive oxygen species may be possible mediators. The present study investigated whether low doses of exogenous hydrogen peroxide could mimic preconditioning in isolated, Langendorff perfused rat hearts. Hearts were subjected to two episodes of 3 min global ischaemia and 5 min reperfusion (n = 17), or were given 10 (n = 15), 20 (n = 10), 30 (n = 20), 40 (n = 18), 80 (n = 17) or 160 microM (n = 10) hydrogen peroxide for 10 min, followed by 10 min recovery, before 25 min global ischaemia and 60 min reperfusion, and compared with ischaemic controls of matching perfusion time (n = 17 and n = 23). Cardiac performance was assessed by heart rate, left ventricular systolic, end-diastolic and developed pressures, and coronary flow. Severe reperfusion arrhythmias occurred frequently in control hearts, and was attenuated by ischaemic preconditioning. All hearts pretreated with 160 microM hydrogen peroxide had severe arrhythmias throughout reperfusion, while these were not seen in any heart perfused with 20 microM hydrogen peroxide (P < 0.01 compared to controls). Ischaemia and reperfusion induced a minor decrease in heart rate, left ventricular systolic and developed pressures, and increased end diastolic pressure. Ischaemic preconditioning attenuated the decrease of heart rate and the increase of end-diastolic pressure, and increased coronary flow, while hydrogen peroxide did not significantly attenuate these changes. In conclusion, a low dose of exogenous hydrogen peroxide before global ischaemia inhibited severe reperfusion arrhythmias, but had no other protective effects. The present work does not suggest that reactive oxygen species are important mediators of the preconditioning effects on stunning and arrhythmias in the rat heart. PMID- 9401579 TI - Vasomotion in rat diaphragm microcirculation at rest and during stepwise arterial pressure reduction. AB - The effect of haemorrhagic hypotension on the incidence, frequency and relative amplitude of vasomotion in rat diaphragm microcirculation was assessed by laser Doppler flowmetry (LDF). Graded bleeding to four hypotension levels (80, 60, 40 and 30% of the control state) were performed in 24 Sprague-Dawley rats. The incidence of vasomotion was 83% in the control state, 96% at the 80% level, 100% at the 60% level, 96% at the 40% level, and 46% at the 30% level. The median fundamental frequency of vasomotion determined manually during the control state and at the hypotension levels (in descending order) was 4.11 (range, 3.29-5.58) cycles min-1 (cpm), 4.48 (3.21-5.92) cpm, 4.20 (3.5-5.56) cpm, 4.01 (3.33-5.36) cpm, 3.71 (3.25-4.49) cpm (P < 0.01 from the fundamental frequency at 80 and 60% hypotension levels). The median relative amplitudes determined manually during the control state and descending hypotension levels were 44.5% (range, 24.9 135.9%), 69.4% (26.6-147.2%), 84.0% (40.3-177.1%) (P < 0.01 from resting and last stage of bleeding), 90.40% (26.2-189.6%) (P < 0.01 from resting and last stage of bleeding), 69.2% (35.6-93.2%). We concluded first that during the resting condition, vasomotion was frequently present in diaphragm microcirculation, which is distinct from other vascular beds of skeletal muscles. Second, the relative amplitude of vasomotion during haemorrhagic hypotension plotted against decreasing blood pressure exhibited a reverse U-shaped curve with a maximum at 40 60% of the control blood pressure, while the frequency of vasomotion remained relatively constant until the last stage of haemorrhage and centred around 3-5 cpm. PMID- 9401580 TI - Acute microcirculatory changes after scalding of the rat paw. AB - A scalding model in the anaesthetized rat was used to measure acute circulatory reactions after heat exposure. Local blood flow of both hindpaws was recorded simultaneously and continuously by laser Doppler flowmetry before, during and for 2 hours following scalding. The scalding injury was inflicted by dipping the right hindpaw into hot water at 60 degrees C for 20 s. Concomitantly, the mean arterial blood pressure (MAP) was displayed on a chart recorder. MAP was obtained by cannulation of the common carotid artery. Oedema formation was calculated by measuring the volume changes of the hindpaws in a plethysmometer before and 30, 60 and 120 min after scalding. Scalding was followed by a biphasic increase of cutaneous circulation. During the first minute after heat provocation, an immediate increase in blood perfusion of about 400% was recorded, followed by a slow decrease of circulation. At 30 min after scalding, there was a secondary phase of increased microcirculation of approximately 230%. A slow decline of cutaneous circulation then followed, and after about 60 min the value was stabilized at approximately 100% above pre-burn level throughout the observation time. Almost no change of perfusion was observed on the contralateral unscalded paw. The scalding injury was followed by a progressive oedema formation on the scalded paw, measured by a volume increase of approximately 72% during the observation period, whereas the non-scalded paw showed no change. MAP remained at a stable level throughout the experiment except for a short-lasting transient increase of approximately 10% at the same time as the first peak of blood perfusion. We could thus confirm that scalding in the present model is accompanied by an immediate and marked increase in the peripheral circulation of the scalded paw followed by a later propagation of oedema, and that these inflammatory changes do not appear to be related to central haemodynamic alterations. PMID- 9401578 TI - Effects of stimulation and inhibition of protein kinase C on the cytosolic calcium concentration in rabbit afferent arterioles. AB - The effect of the protein kinase C (PKC) inhibitor chelerytrine (Ch) and the PKC activator 12-0-tetradecanoyl-phorbol-13-acetate (TPA) on the cytosolic calcium concentration ([Ca2+]i) in isolated intact rabbit afferent arterioles was investigated. [Ca2+]i was measured in the proximal and distal parts of the arteriole. Administration of 1 microM Ch gave rise to a peak followed by an elevated level of [Ca2+]i in both these parts. Neither the peak nor the elevated level of [Ca2+]i was significantly reduced by 1 microM nifedipine. The relative peak increase in [Ca2+]i in response to 1 microM noradrenaline (NA) or to 10 nM angiotensin II (AII) was significantly blunted in both parts after preincubation with 1 microM Ch. Depolarization with 25 mM K+ increased [Ca2+]i in both parts. Preincubation with Ch did not affect the increase in [Ca2+]i induced by 25 mM K+. TPA (10 and 100 nM) did not significantly affect the basal [Ca2+]i in the afferent arteriole. The [Ca2+]i response to NA or 25 mM K+ was not affected by TPA. We conclude that blockade of PKC increases [Ca2+]i in afferent arteriolar smooth muscle by a mechanism independent of L-type voltage-sensitive calcium channels. Inhibition of PKC blunts the relative increase in [Ca2+]i in response to AII and, to a lesser extent, that induced by NA. We conclude that PKC might be important in modulating the calcium changes that occur in response to these vasoconstrictors. PMID- 9401581 TI - Radiotelemetrically recorded blood pressure and heart rate changes in relation to plasma catecholamine levels during parturition in the conscious, unrestrained goat. AB - The aim of this study was to investigate the extent of sympathetic nervous system activation during parturition in four unrestrained goats. Chronically implanted radiotelemetry devices registered heart rate and arterial blood pressure around the clock and blood was sampled for determination of plasma adrenaline and noradrenaline concentrations before, during and after labour. Two goats delivered two kids after moderately intensive abdominal contractions. A third goat had dystocia, and was treated with prostaglandin F2 alpha. One normal kid and one mummified foetus were delivered manually. After milking, a third kid was born spontaneously. The fourth goat experienced severe abdominal contractions and delivered one kid. Mean blood pressure was 69 +/- 2 mmHg the day before parturition, increased gradually during the labour pains, and reached a maximal value of 120 +/- 7 mmHg when the head of the first kid was visible (P < or = 0.05). Heart rate was 134 +/- 4 beats min-1 the day before parturition and peaked when the first kid was born (159 +/- 6 beats min-1, P < or = 0.05), as did plasma adrenaline concentration (from 0.4 +/- 0.2 nmol L-1 to 2.7 +/- 1.2 nmol L-1, P < or = 0.05). The concentration of noradrenaline increased from 4.8 +/- 2.3 nmol L 1 to 12.2 +/- 8.4 nmol L-1 (P < or = 0.05), when the head of the first kid was visible. Expulsion of the second and third kids caused relatively smaller increases in blood pressure, heart rate and catecholamines than those seen with the first born kid. It is concluded that changes in pressure, heart rate and catecholamines during parturition are related to the different phases of labour and not to its duration or severity. PMID- 9401582 TI - Importance of nitric oxide in hepatic arterial blood flow and total hepatic blood volume regulation in pigs. AB - The importance of nitric oxide in regulating basal arterial blood flow has been examined in several different vascular beds by intra-arterial infusion of inhibitors of nitric oxide synthesis, but not in the arterial vascular bed of the liver. In the present study, N(G)-nitro-L-arginine (L-NNA), in a dose of 0.5 and 1.0 mumol mL-1 of hepatic arterial blood flow, was infused for 5 min into the hepatic artery in seven pigs anaesthetized with pentobarbital sodium. The haemodynamic effects observed by the first infusion were not further enhanced by the second infusion. Hepatic arterial resistance increased by 143 +/- 38% and hepatic arterial blood flow declined by 38 +/- 10%. A systemic effect due to 'spillover' was observed, as evidenced by an increase in mean aortic blood pressure of 24 +/- 4 mmHg. However, no significant increase in arterial mesenteric resistance was observed and total liver blood flow remained unchanged. Hepatic arterial vasodilation in response to occlusion of the portal vein, the arterial buffer response, remained intact after inhibition of nitric oxide synthesis. Liver lobe thickness, measured by an ultrasonic technique, was not found to change with inhibition of arterial nitric oxide synthesis, excluding a significant direct effect of arterial nitric oxide on liver capacitance. In conclusion, nitric oxide is an important regulator of hepatic arterial resistance, but does not mediate the hepatic arterial buffer response and was not found to play any significant role in total hepatic capacitance regulation. PMID- 9401583 TI - Effect of inhaled nitric oxide on endotoxin-induced hypoxaemia in rabbits. AB - In five mechanically ventilated rabbits, we studied the property of inhaled nitric oxide in helping to treat hypoxaemia which was induced by intravenous endotoxin (Escherichia coli-derived lipopolysaccharide, serotype 0111: B4). We used measurements of arterial partial pressure of oxygen to check a therapeutic nitric oxide benefit. Pulmonary artery pressure was continuously monitored. Furthermore, we determined the single-breath diffusing capacity for nitric oxide. Measurements of plasma nitrite/nitrate concentration served as an indicator of endogenous nitric oxide output. The first infusion of endotoxin led to a transient pulmonary vasoconstriction, whereas arterial partial pressure of oxygen was permanently reduced by 30 +/- 10 mmHg (mean +/- SD), attaining minimal values of 48 +/- 3.4 mmHg due to additional endotoxin. Single-breath diffusing capacity for nitric oxide declined by 20 +/- 5.5% of baseline values until the experiments were concluded. Endotoxin induced an increase in plasma nitrite/nitrate concentration in the five rabbits as well as in the control animals (four rabbits) without exogenous nitric oxide supply. During the 25 inhalations of nitric oxide (3-50 ppm), arterial oxygenation did not change significantly. Thus endotoxin permanently impaired pulmonary gas exchange without inducing pulmonary hypertension. Inhaled nitric oxide did not improve arterial oxygenation during endotoxaemia. PMID- 9401584 TI - Effects of the beta 2-adrenergic agonist clenbuterol on capillary geometry in cardiac and skeletal muscles in young and middle-aged rats. AB - The effects of 10 day clenbuterol administration on cardiac and skeletal muscle capillarities were studied, particularly in terms of the distribution of arteriolar and venular capillaries and their capillary density, in young (10-week old) and middle-aged (37-week-old) male Wistar rats. Rats of the treated groups were fed a diet containing 2 mg kg-1 clenbuterol hydrochloride. In both young and middle aged rats, clenbuterol treatment increased the body wt and the weights of the heart and hindlimb muscles. The mean fibre cross-sectional area was significantly increased after the treatment in the left ventricle, soleus, plantaris and both deep and superficial portions of gastrocnemius (P < 0.01). In the left ventricle, the total capillary density and the density of venular capillaries were decreased after the treatment in both young (9 and 13%, respectively) and middle-aged rats (10 and 11%, respectively). A decrease in total capillary density was also observed in all skeletal muscles examined. In both young and middle-aged rats, the capillary-to-fibre (C:F) ratio and the proportion of each capillary did not change after the treatment in both the left ventricle and skeletal muscles. Clenbuterol significantly decreased the activity of succinate dehydrogenase in all skeletal muscles examined (P < 0.01). These results suggest that clenbuterol increased the diffusion distance for oxygen in the left ventricle and skeletal muscles. These changes may reduce the oxygen supply to tissues and increase muscle fatigability. PMID- 9401585 TI - Time-dependent effects of ischaemia on neuropeptide Y mechanisms in pig renal vascular control in vivo. AB - We have investigated the effects of ischaemia on neuropeptide Y (NPY) mechanisms involved in sympathetic vascular control of the pig kidney in vivo. Reperfusion after 2 h of renal ischaemia was associated with local overflow of noradrenaline (NA) but not of NPY-like immunoreactivity (-LI). Renal sympathetic nerve stimulation 10 min into reperfusion evoked markedly reduced vasoconstrictor effects and significantly less overflow of NA (reduced by 70% from the pre ischaemic conditions), whereas NPY-LI overflow was unaltered. Renal vasoconstrictor responses to exogenous peptide YY (PYY), phenylephrine and angiotensin II were strongly attenuated after this ischaemic period, while vasoconstriction to alpha, beta-methylene ATP was maintained to a larger extent. The renal vascular responses and NA overflow had become partially normalized within a 2 h recovery period. In contrast, the renal vasoconstrictor response and the overflow of NPY-LI upon sympathetic nerve stimulation were enhanced after 15 min of renal ischaemia. In parallel, the PYY-evoked renal vasoconstriction was selectively and markedly prolonged after the 15 min of ischaemia. In the presence of the NPYY1 receptor antagonist BIBP 3226, the augmented vascular response to nerve stimulation was significantly attenuated. We conclude that reperfusion after 2 h of renal ischaemia is associated with local overflow of NA, whereas the sympathetic nerve-evoked release of NA and the reactivity of the renal vasculature to vasoconstrictor stimuli are reversibly reduced. Furthermore, possibly due to an impaired local degradation, the role of neurogenically released NPY in renal sympathetic vasoconstriction is enhanced after short-term (15 min) ischaemia compared with control conditions. PMID- 9401586 TI - Effects of aminoguanidine and L-NAME on histamine-induced blood pressure drop in the rat. AB - Mean arterial blood pressure changes in response to i.v. administration of histamine were monitored in the anaesthetized rat in the absence or presence of the diamine oxidase (DAO) inhibitor aminoguanidine (AMG, 10 mg kg-1). AMG prolonged the duration of the transient drop in blood pressure induced by a bolus injection of histamine (0.05 mg kg-1) by 34%. In animals pretreated with AMG, no potentiation of the decrease in pressure in response to a 10 min infusion of histamine was observed. However, when infusion was stopped, the time needed for pressure recovery was twice as long in animals treated with AMG as in controls. Blood samples were taken prior to infusion and during the recovery phase and the quantities of histamine were determined by liquid chromatography. The prolonged recovery phase observed in animals pretreated with AMG was associated with five times higher levels of histamine. The duration of histamine-induced hypotension (0.01 mg kg-1) was 50% shorter in the presence of the nitric oxide synthase inhibitor L-NAME (10 mg kg-1). We suggest that DAO, through elimination of histamine from the bloodstream, is important for the recovery from histamine induced hypotension, and that the duration of histamine-induced pressure drop is influenced by formation of nitric oxide. PMID- 9401587 TI - Rate of oxidative phosphorylation in isolated mitochondria from human skeletal muscle: effect of training status. AB - Muscle oxidative function has been investigated in subjects with various training status (VO2 max, 41-72 mL O2 kg-1 body wt min-1, n = 10). Mitochondria were isolated from biopsies taken from m. vastus lateralis. Maximal mitochondrial oxygen consumption (QO2) and ATP production (MAPR) were measured with polarographic and bioluminometric techniques, respectively. The yield of mitochondria, calculated from the fractional activity of citrate synthase (CS), averaged 26%. With pyruvate + malate, the respiratory control ratio was 5.7 +/- 0.4 (X +/- SE) and the P/O ratio was 2.83 +/- 0.02, which demonstrates that the isolated mitochondria were functionally intact. QO2 was significantly correlated to aerobic training status expressed as muscle CS activity (r = 0.86), VO2 max (r = 0.84) and lactate threshold (r = 0.83) but not to the fibre type composition. A highly significant correlation (r = 0.93) was observed between ATP production calculated from QO2 and MAPR, but ATP production derived from QO2 was higher than MAPR both for pyruvate + malate (255%) and for alpha-ketoglutarate (23%). QO2 extrapolated to a temperature of 38 degrees C averaged 68 mL O2 min-1 kg-1 wet wt, which is similar to previous findings in vitro and in vivo during the post exercise period. However, calculated muscle O2 utilization during exercise was three- to fivefold higher than QO2 measured on isolated mitochondria. It is suggested that additional factors exist for activation of mitochondrial respiration during exercise. It is concluded that muscle oxidative function can be quantitatively assessed from the respiration of mitochondria isolated from needle biopsy specimens and that QO2 is closely correlated to whole-body VO2 max. PMID- 9401588 TI - Central and peripheral components of diaphragmatic fatigue during inspiratory resistive load in cats. AB - The development of fatigue was investigated in the diaphragm of anaesthetized, tracheostomized, spontaneously breathing cats during restricted air flow. Ventilation, transdiaphragmatic pressure (Pdi), integrated electrical activity of diaphragm (Edi) and phrenic nerve (Eph) were measured simultaneously and expressed as a percentage of values at unloaded breathing. Inspiratory loads were 60, 70 and 80% of Pdi max. The Pdi max was measured by airway occlusion at functional residual capacity. The duration of loads was 40-60 min. The diaphragmatic fatigue developed only during heavy inspiratory loading (80% Pdi max). During the first 10 min of heavy load Pdi, Edi and Eph increased to 905 +/- 60%, 248 +/- 20% and 229 +/- 24%, respectively (P < 0.01), and then began to fall gradually. Ventilation declined to 39 +/- 3% after 60 min of heavy load (P < 0.01), resulting in acute hypercapnia and hypoxia. Initial fatigue appeared as a decrease in Pdi (to 781 +/- 63%) and parallel decline in Edi (to 233 +/- 21%) after 30 min of load (P < 0.05). Phrenic nerve activity did not change during this stage. These data suggest a peripheral basis of diaphragmatic fatigue, related to disorders in neuromuscular transmission. After 60 min of heavy load, Pdi fell to 675 +/- 49%, Edi declined to 209 +/- 28% and Eph decreased to 189 +/- 25%. We interpret the decrease in phrenic nerve activity as a weakening of central inspiratory drive and development of the central component of diaphragmatic fatigue in the last stage. PMID- 9401589 TI - Intramuscular EMG from the hip flexor muscles during human locomotion. AB - The purpose was to investigate the activation pattern of five major hip flexor muscles and its adaptation to changing speed and mode of progression. A total of 11 healthy subjects performed walking and running on a motor-driven treadmill at speeds ranging from 1.0 to 6.0 m s-1. Intramuscular fine-wire electrodes were used to record myoelectric signals from the iliacus, psoas, sartorius, rectus femoris and tensor fascia latae muscles. The basic pattern, with respect to number of activation periods, remained the same irrespective of speed and mode of progression. However, differences in the relative duration and timing of onset of activation occurred between individual muscles. Over the speed range in walking, a progressively earlier onset was generally seen for the activation period related to hip flexion. Changes in EMG amplitude were measured in the iliacus and psoas muscles and showed a marked increase and difference between walking and running at speeds above 2.0 m s-1. Thus, the alternating flexion-extension movements at the hip during locomotion appear to be governed by a rather fixed 'neural program' which normally only needs minor modulations to accomplish the adjustments accompanying an increase in speed of progression as well as a change from walking to running. PMID- 9401590 TI - Influence of muscle mass and work on post-exercise glucose and insulin responses in young untrained subjects. AB - The 18 h post-exercise glucose and insulin responses of six male and six female subjects were measured following one- or two-leg cycling to determine the influence of muscle mass involvement and work. Each subject performed three exercise trials on a Cybex Met 100 cycle ergometer: (1) two-leg exercise for 30 min at 60% of the two-leg VO2 max; (2) one-leg exercise for 30 min at 60% of one leg VO2 max; and (3) one-leg exercise (one-leg TW) at 60% of the one-leg VO2 max with the total work performed equal to that of the two-leg trial (duration approximately 50 min). These trials were preceded by 2 days of inactivity and followed by an 18 h post-exercise 75 g oral glucose tolerance test (OGTT). The glucose response during the baseline OGTT demonstrated that the subjects had normal glucose tolerance with fasting serum glucose levels of 5.1 mM, and 1 and 2 h serum glucose less than 7.8 mM, respectively. The 18 h post-exercise glucose responses were significantly lower following the two-leg trial (P < 0.05), with the area under the curve values being 129.9 mM h-1 less than the resting control level. The 18 h post-exercise insulin AUC response of the two-leg trial was significantly lower than either of the one-leg responses (14.7 pM below the one leg and 5.0 pM below the one-leg TW) but was not associated with a change in C peptide. The 18 h post-exercise insulin levels of the one-leg and one-leg TW trials were above or near the resting control values, but were not accompanied by a significant change in C-peptide. In conclusion, the data presented here show that the amount of muscle tissue utilized during an exercise bout can influence both the glucose and insulin responses, whereas the amount of total work employed during the exercise had no effect on either of these parameters. PMID- 9401591 TI - Effects of ubiquinone-10 supplementation and high intensity training on physical performance in humans. AB - This study investigated the effects of oral supplementation with ubiquinone-10 (Q10) (n = 9) compared with a placebo (n = 9) on aerobic and anaerobic physical performance over 22 days of supplementation. The supplementation period included 5 days of high intensity anaerobic training between days 11 and 14. The results demonstrated, that on an anaerobic (10 x 10 s) cycling test, the placebo group showed a significantly greater improvement than the Q10-group after a supplementation and training period (P < 0.001). Further, the Q10 group had a significantly lower increase in total work performed during the seven training sessions (15 x 10 s) compared with the placebo group (P < 0.001). There was a significant increase in maximal blood lactate accumulation during cycling in the both groups, when compared with levels before the training and recovery period. There was no significant difference between the groups, either in VO2max determined during running, or in submaximal and peak VO2, Rate of Perceived Exertion, respiratory quotient, blood lactate concentration or heart rate determined during submaximal and maximal cycling. Although insignificant (P = 0.1 0.3), there was evidence of higher submaximal VO2 (55-80% of VO2peak) during cycling in the Q10-group compared with the placebo group after training and recovery. It is concluded that with high intensity anaerobic training, there was a significantly greater increase in anaerobic performance in the placebo group compared with the Q10 group. The results suggest less increase in physical performance with Q10 supplement and high intensity anaerobic training, compared with placebo. PMID- 9401592 TI - Blubber and flipper heat transfer in harp seals. AB - The trunk of marine mammals is encased in a blubber layer which provides thermal insulation that can be changed by circulatory adjustments. The extremities, on the other hand, are poorly insulated but have vascular arrangements constructed for prevention or promotion of heat loss depending on the thermal state of the animal. We have studied the importance of different body parts as sites for heat dissipation and also assessed the effect of circulatory adjustments on heat transfer through blubber, by combining direct measurements of heat flux from the flippers and trunk with simultaneous recordings of temperature gradients through the blubber and metabolic rates of harp seals (Phoca groenlandica) subjected to water temperatures between 1 and 24 degrees C. We also determined the thermal conductivity of blubber samples from the same animals after death, and compared this with the insulative properties of live blubber. At the lowest water temperatures, the insulative properties of live blubber were similar to those of dead blubber, and heat loss from the flippers only accounted for 2-6% of the metabolic heat production. As heat load increased with increasing water temperatures, the fraction of heat lost from the flippers increased, to 19-48% at 24 degrees C, while the fraction lost from the trunk decreased, despite an increase in the convective (circulatory) heat transfer through the blubber layer. PMID- 9401593 TI - Circadian variation of sweating responses to passive heat stress. AB - The aim of present study was to examine whether sweating responses to passive heat stress change with the circadian rhythm of internal temperature. Six men had their legs immersed in water at 42 degrees C for 60 min in an ambient temperature of 28 degrees C on four separate days. Experiments were conducted at four different times [06.00 h (morning), 12.00 h (daytime), 18.00 h (evening) and 24.00 h (night)]. We measured oesophageal temperature (Toes), mean body temperature (Tb), local sweating rate (msw) on the forehead, back, forearm and thigh, the densities of activated sweat gland (ASG) on the back, forearm and thigh, and the frequency of sweat expulsion per minute (Fsw) which has been suggested to represent central sudomotor activity. Sweat gland output (SGO) on each site was calculated by dividing msw by ASG. ASG was significantly higher on the forearm than on the back and thigh, and SGO was significantly lower on the forearm than on the back and thigh. However, ASG and SGO did not significantly change over the day. Tb and Toes thresholds for the onset of sweating showed a significant change with both the temperature rhythms at rest prior to each procedure, while the slopes of the relationships Fsw-Tb and msw-Fsw showed no significant difference over the day. We suggest that the circadian variation of sweating response to passive heat stress is regulated by a central sudomotor mechanism rather than by sweat gland function. PMID- 9401594 TI - Effects of pretreatment with an indeno-indole compound on lipid peroxidation in the cortex and medulla of rabbit kidneys after ischaemia-reperfusion. AB - The effects of 60 min of ischaemia with or without reoxygenation in vivo or in vitro on lipid peroxidation in cortical and medullary tissue from rabbit kidneys were measured as production of thiobarbituric acid-reactive substances (TBARS). Lipid peroxidation was more pronounced in medullary tissue compared with cortical tissue. The highest TBARS production was found in medullary slices subjected to reoxygenation in vitro immediately after 1 h of ischaemia. Reperfusion in vivo before reoxygenation in vitro attenuated the TBARS formation during subsequent in vitro incubation. Pretreatment of the rabbits with an indeno-indole compound (code name H 290/51) reduced the TBARS formation after 60 min of ischaemia and reoxygenation in vitro towards control values. PMID- 9401595 TI - CGRP, but not substance P, induces an increased negativity of the interstitial fluid pressure in rat trachea. AB - Neurogenic inflammation is mediated by neuropeptides released from sensory nerves following electrical stimulation of the vagal nerve or by capsaicin. The released neuropeptides are, among others, calcitonin gene-related peptide and substance P, which both induce vasodilation, while only substance P induces plasma extravasation. Electrical stimulation of the vagal nerve induces increased negativity of interstitial fluid pressure (Pif), which will contribute to enhance oedema formation. Pif was measured, on the abluminal side of the surgically exposed trachea, with sharpened glass capillaries (4-10 microns) connected to a servo-controlled counterpressure system. Measurements were performed after circulatory arrest, since the oedema formation associated with acute inflammation will increase Pif in a positive direction, which may potentially underestimate the increased negativity of Pif. Experiments were carried out in pentobarbital anaesthetized (50 mg kg-1) Wistar-Moller rats. Pif in control rats averaged -1.2 +/- 0.9 (SD) mmHg (n = 9). Intravenous injection of capsaicin (65.0 nmol) and calcitonin gene-related peptide (1.3 nmol) increased the negativity of Pif to 4.0 +/- 1.2 mmHg (n = 8) (P < 0.01) and -4.7 +/- 2.0 mmHg (n = 9) (P < 0.01), respectively. Intravenous injection of substance P (7.4 nmol, n = 9; and 37.0 nmol, n = 8) did not affect Pif compared to control (P > 0.05). Similarly, potentiation of the available substance P with thiorphan or captopril did not increase the negative Pif, nor did injection of stable substance P analogues. Thus, the present study seems to support the theory that, in rat trachea, the increased negativity of Pif after intravenous injection of capsaicin and after vagal stimulation is caused by calcitonin gene-related peptide. PMID- 9401596 TI - The activation pattern in normal humans during suppression, imagination and performance of saccadic eye movements. AB - The distribution of activated cerebral regions was examined in nine normal subjects during four different eye movement-related conditions: (1) fixation fixation on a central light emitting diode; (2) saccadic suppression-fixation on a diode in the presence of flashing lateral targets; (3) reflexive/volitional saccades-performance of overt eye movements to two laterally lit targets and back to the centre; and (4) imagined saccades-imagining, but not performing, the same eye movements. The regional neural activity was measured indirectly using repetitive bolus injections of oxygen-15-labelled water and positron emission tomography (PET) to yield time-integrated images of the normalized count distribution. These were aligned and anatomically normalized to a standard stereotactic space and the averages of each condition were analysed categorically using statistical parametric mapping. Compared to central fixation, reflexive/volitional saccades significantly activated regions in the classically known cortical oculomotor regions. The most notable activation during the saccade suppression task, compared to central fixation alone, was a bilateral activation of the parietal cortex with a right-sided preponderance, activation of the supplementary eye field/caudal cingulate regions, and activation of frontal regions close to the frontal eye fields. Imagined performance of eye movements without overt eye movements activated the supplementary eye field and frontal eye fields identically to regions involved in overt eye movements, thus demonstrating that over eye movements are not a prerequisite of the activation of these regions in normal humans. PMID- 9401597 TI - Increased activity of citrate synthase in human skeletal muscle after a single bout of prolonged exercise. PMID- 9401598 TI - James Paget Henry. 12/7 1914--20/11 1996. PMID- 9401599 TI - Psychological and physiological responses to stress: the right hemisphere and the hypothalamo-pituitary-adrenal axis, an inquiry into problems of human bonding. AB - In addition to repeated reexperiencing of the event, the delayed effects of severe psychological trauma, i.e., post traumatic stress disorder (PTSD), present a paradoxical mix of symptoms. There is enhancement of the self-preservative catecholamine states; anger and fear with a contrasting sense of meaninglessness and blunting of the emotional responses of the attachment behavior so critical for species preservation. Hormonally, there is a striking separation of the catecholamine response, which stays elevated and that of the hypothalamo pituitary-adrenal (HPA) axis, which may remain at normal levels. Pathophysiologically, the reexperienceing of the trauma and the arousal may be associated with dysfunction of the locus coeruleus, amygdala and hippocampal systems. This article explores the consequences of an additional dysfunction: a dissociation of the hemispheres that appears to be responsible for the alexithymic avoidance and failure of the cortisol response that so often follow severe psychological trauma. There is neurophysiological evidence that the left the right hemispheres subserve different emotional sets that correspond to "control" and "appraisal," i.e., very approximately to the self and species preservative behavioral complexes, respectively. Several studies point to physiological dissociation of hemispheric functions during alexithymia. This raises the question: What has been lost if in this condition the right side no longer fully contributes to integrated cerebral function? Right hemispheric damaged children lose critical social skills and in adults the related sense of familiarity critical for bonding is lost. Such losses of social sensibilities may account for the lack of empathy and difficulties with bonding found in sociopathy and borderline personality: conditions now believed to result from repeated psychological trauma during development. On the other hand, systems that promote right hemispheric contributions provide solacing access to a "Higher Power." They also appear to protect against socially disordered behavior, substance abuse, the failure of the HPA axis and some aspects of the pathophysiology of chronic disease. PMID- 9401600 TI - James Paget Henry--a retrospective. AB - James Paget Henry really began his productive research career at the outset of the second world war. His studies of acceleration and the anoxia of high altitude were supported by the development of then new techniques of measuring and recording critical physiologic parameters such as vascular pressures, respiratory functions and haemoglobin saturation. His inquisitive mind made productive use of the instruments that had to be made by skilled instrument makers working in university shops. Much of this instrumentation has now found its way into the clinical arena where it is now the main armamentarium of cardiac diagnostic and respiratory function laboratories. His work in the space program preceeded that of the Russians but did not get recognition until Sputnik awakened the world to the possibilities of space flight. His development of the concept of a cardiovascular basis for fluid volume control and the supportive investigative work undertaken constitute a milestone in the annals of experimental physiology. The chimpanzees used in Project Mercury were found to be hypertensive which was related to the method of capture used by the commercial suppliers. This lead Jim to study the effect of early experience on resting blood pressure, an effort that soon developed into provocative studies of the biological basis of the stress response. PMID- 9401601 TI - Shifting the circulatory control paradigm. AB - The current paradigm regards circulatory control as mediated by discrete cardiovascular receptor stimuli, through an array of relatively independent reflexes, with the arterial baroreflex the centrepiece of this schema. However, it is often difficult to fit the linear control model to the observed responses of the intact organism. Hence the need for a more realistic approach. A given disturbance acting on the body stimulates not only baroreceptors, but many other receptors, as well as providing the central nervous system (CNS) with behavioral cues, etc. The mix of stimuli is characteristic of the type and severity of the particular disturbance. The first task for the CNS, is recognition of the pattern of stimuli, which is often a non-linear process. This is mediated through a number of "integrative" centres in different parts of the brain, which compare the magnitude of stimuli from the various sources. The sum of excitatory and inhibitory influences in the efferents from these integrative centres project to "command" centres, e.g. in hypothalamus, amygdala or hindbrain. These generate the output patterns through activation of particular pools of autonomic motoneurons and by altering secretion of hormones. Both the recognition of afferent stimuli (including behaviour) and of the effector patterns, involve mechanisms above and below the pons. PMID- 9401602 TI - Physiological aspects of the "defence" and "defeat" reactions. AB - By means of tele-receptor signals (vision, hearing, olfaction) the mammalian brain is almost continuously informed about environmental events, and whenever these are interpreted as positive or negative challenges the cerebral "super controller" can, for coping with the anticipated situation, select the most appropriate among a number of pre-formed hypothalamic reaction patterns. These are organized as combined engagements of the somatomotor, visceromotor and hormonal efferent links, whereby a variety of behavioural responses can be elicited, where each is accompanied by appropriate adjustments of inner organ systems, metabolism, etc., to achieve optimal performance. For eons of time these "emotionally charged" reactions, common for all mammals, have served to protect the individual and species in a merciless environment, and they certainly remain principally the same also when Homo Sapiens faces modern society. As then the ancient "defence" and "defeat" reactions, intended for quite different situations, are often activated by the many artificial stimuli and symbolic threats inherent in today's hectic and competitive life, their principal organisation and functional consequences are the main topic of this survey. They are, for example, also marginally engaged along with ordinary shifts in mood during events in daily-life but are probably in this mild form fairly harmless and actually often supportive for efficient performance. However, when intensely engaged over longer periods they can, indeed, profoundly disturb inner organ systems and metabolic events, often resulting in disorder and even in premature death, as particularly convincingly shown by Henry and co-workers in studies on rodent "micro-societies". Transferred to man's situation in modern life, these model studies have been crucial for insight into the indeed complex mechanisms involved when long-term psychosocial stress in predisposed or particularly exposed individual contributes to some of today's most important "disorders of civilisation". PMID- 9401603 TI - Oxytocin linked antistress effects--the relaxation and growth response. AB - Stress or noxious stimuli of various kind may induce the fight-flight response. In this situation a number of physiological and behavioural adaptations leading to defense of the organism occur. At a central level increased activity in the noradrenergic locus coeruleus (LC) and an enhanced secretion of corticotrophin releasing factor (CRF) and vasopressin produced in the paraventricular nucleus (PVN) integrate stress response. Here the existence of an opposite psycho physiological pattern associated with relaxation and growth and which is activated by certain types of non-noxious stimuli and integrated by oxytocin is proposed. In support of this, administration of oxytocin to male and female rats gives rise to effects of antistress nature in particular after repeated administration. Thus a five day treatment period with oxytocin 1 mg/kg s.c. or 1 micro g/kg i.c.v gives rise to sedation, lowering of blood pressure, increased withdrawal latency in the tail flick test and also a decrease of corticosterone levels and a rise of certain vagally controlled hormones. Weight gain is also increased under certain conditions. These effects persist several weeks after administration of oxytocin and cannot be reversed by oxytocin antagonists when established, suggesting that secondary mechanisms have been activated. Naloxone temporarily reverses the increased withdrawal of the tail flick test suggesting that opioid mechanisms have been activated to cause this particular effect. In contrast the sedative and blood pressure lowering effect seems to be induced by an enhanced activity in central alpha 2 receptors. Oxytocin levels increase in blood and CSF after various kinds of non-noxious sensory stimulation such as touch, light pressure and warm temperature in both female and male rats. It is suggested that other types of non-noxious stimuli as well may increase oxytocin release. If so, a release of oxytocin could be responsible for not only the antistress effects occurring during lactation but also why relationships, social contact and networks may have health promoting effects in particular by preventing cardiovascular disease. PMID- 9401604 TI - Stress and immunity: what have we learned from psychoneuroimmunology? AB - The old concept that stress depresses immunity must be qualified. There is now evidence that in the same way that different perceptions of stress have different physiological consequences, different ways of coping with stress result in different consequences on immunity, the nature and outcome of which depend on the type of immune response. The mechanisms that are involved in these effects involve neuroendocrine and autonomic pathways. These pathways are actually part of a network of bidirectional interactions between the central nervous system and the immune system, which plays an important role in the physiological regulation of immunity. PMID- 9401605 TI - Physical exercise, endogenous opioids and immune function. AB - The experimental data available today strongly indicate that various types of physiological stressors, including physical exercise and emotional stress, can influence immune function. Natural immunity represents a first line of defence in viral infections and cytotoxicity to a variety of tumour cells. Natural immunity is strongly influenced by chronic exercise and this regulation includes interaction between the nervous, endocrine and immune systems. Central mechanisms including the endogenous opioids are of great interest. Chronic activation of endogenous opioid systems augments natural cytotoxicity and the possible involvement the opioids in the exercise-induced enhancement of natural immunity is discussed. Also, catecholamines seem to play an important role in the regulation of immune function, both after chronic exercise and emotional stress. The physiological significance of the reported changes in natural cytotoxicity after exercise-training is as yet unclear. PMID- 9401607 TI - The right hemisphere and the human stress response. AB - Evidence is presented that most components of the human stress response are influenced asymmetrically by the cerebral hemispheres. The right hemisphere is endowed with a unique response system preparing the organism to deal efficiently with external challenges. Therefore, both the hypothalamic-pituitary adrenocortical (HPA) axis and the sympathetic-adrenomedullary (SA) axis seem to be under the main control of the right hemisphere. PMID- 9401606 TI - The role of the central amygdala in stress and adaption. AB - Recent views on stress emphasise the existence of more than one response pattern to stressful events, and the importance of individual differences in coping with environmental challenges. Therefore, in the evaluation of the specific contribution of the central nucleus of the amygdala (CEA) in stress and adaptation it has to be considered whether the stress is acute or conditioned, and whether it requires active or passive styles of coping. Based on series of our own studies, we propose that the CEA is consistently involved in the organisation of processes of passive coping, reflected in immobile behaviour and parasympathetic activity. Furthermore, a differential regulation of the CEA via its peptidergic neuronal input may underlie the distinct behavioural and physiological stress patterns accompanying the different styles of coping. Additionally, the involvement of the CEA in neuroendocrine control is addressed. The CEA exerts a general, modulatory influence on the neuroendocrine control to acute stressors, whereas this output during conditioned stress seems to be independent of the CEA. The neuroendocrine state as achieved during acute stress is, via feedback to the brain, of importance in learning about the situation, and to consolidate the experience. In this review an attempt is made to provide an integrated model of CEA functioning in relation to stress and adaptation. PMID- 9401608 TI - Cerebral laterality, repressive coping, autonomic arousal, and human bonding. AB - Jim Henry wrote extensively about emotional expressive styles, such as alexithymia which is characterized by reduced awareness of one's own or others' feelings and emotions, and their relation to cerebral hemispheric asymmetries. The repressive coping style is a stable individual characteristic, which is marked by reduced and minimized reports of stress coupled with higher levels of autonomic, somatic, and behavioural responsivity. The apparent dissociation between subjective and physiological response may be associated with a functional disconnection between the two cerebral hemispheres and with greater cerebral lateralization. To test this hypothesis, we reexamined data from a study in which emotional and neutral slides were presented unilaterally to the left and right hemisphere. Exposure duration was 200 ms. Subjects were divided into four different coping styles based on their defensiveness and anxiety scores. Repressive copers were the only group to show a significant cardiac response (heart rate deceleration) to emotional material when it was presented to the right but not to the left hemisphere. These findings and the fact that repressive copers have a high need for social approval support Henry's views about the role of the right hemisphere in affiliation and human bonding. PMID- 9401609 TI - Genetic influences on the responses to psychosocial challenges in rats. AB - "Psychosocial stress can induce chronic hypertension in normotensive strains of rats" (Henry et al. 1993). This effect is however variable among different strains. Factor analysis of the relationships between temperamental dimensions measured in social and nonsocial settings shows that sensitivity to chronic social stress loads on a "social reactivity" factor different from spontaneous aggressive tendencies. Furthermore an "end-organ" sensitivity of the cardiovascular system also seems to be necessary to explain individual vulnerability to psychosocial stress-induced hypertension. These psychological and pathophysiological characteristics combine with situational components that are not yet fully defined. The advent of telemetry techniques to monitor heart rate and blood pressure continuously without any disturbance to the animals should facilitate the analytical approach of this complex, multifactorial condition. PMID- 9401610 TI - Social stress in rats and mice. AB - This paper summarizes some of the highlights of our current social stress research in rodents as it was inspired by the work of Jim Henry. First, it is argued that social defeat can be considered as one of the most severe stressors among a number of laboratory stressful stimuli in terms of neuroendocrine activation. Moreover, the stress response induced by defeat in particular is characterized by a strong sympathetic dominance. Depending on the stress parameter, the stress response induced by a single social defeat may last from hours to days and weeks. As a long term consequence of a single defeat experience, the animal becomes sensitized to subsequent minor stressors. Finally, the importance of individual differences in coping style in relation to stress vulnerability is discussed. PMID- 9401611 TI - Effects of cage enrichment on territorial aggression and stress physiology in male laboratory mice. AB - The activation of different neuroendocrine subsystems depends on the individual perception and coping with the challenging situation, the formulation of these relations by J.P. Henry represents a most useful concept also for the assessment of welfare consequences of particular caging variables. We investigated effects of cage enrichment on behaviour and neuroendocrine activations of male laboratory mice. Mice in enriched cages behaved more aggressive, lacked stable dominance hierarchies and exhibited neuroendocrine alterations depending on their individual social position. Subdominant passive mice were characterized by an augmented adrenal capacity to synthesize epinephrine despite low activities of the tyrosine hydroxylase. Dominant mice showed elevated circulating corticosterone concentrations despite high tyrosine hydroxylase-activities. Findings showed a dissociation of neurosympathetic and adrenomedullary components in subdominant passive mice and a simultaneous activation of sympathetic adrenomedullary and hypothalamo-adrenocortical components in dominant mice. Within the conceptual framework of the Henry model this would suggest different deteriorations of welfare in dominant and subdominant passive mice. In the situation of intensified aggression in the enriched cages the increased epinephrine synthesis in subdominant mice reflect their more frequent receipt of attacks and the elevated corticosterone secretion in dominant mice reflect their hindered ability to control the dominant position. PMID- 9401612 TI - Social relations and their health impact in tree shrews. AB - In the wild, tree shrews (Tupaia belangeri) live in pairs in territories which they defend vigorously against strange conspecifics. The paper gives an overview on some of our laboratory studies with tree shrews, which demonstrate the great relevance the concepts of Jim Henry have in understanding the biological relevance of mammalian social behavior. A basic result of these studies is that the physiological consequences of social relations between mammals depend on the appraisal of the situation by the animals and their coping behavior. Appraisal of a stimulus or a situation, as well as the resulting coping behavior, are basically psychological processes. There are, therefore, no simple relationships between stimuli imposed on individuals and their physiological responses; rather the behavioral, psychological and, thus, the physiological responses of individuals to stimuli differ depending on their genetics, prenatal influences and postnatal learning processes. This means, to understand the consequences of social relations between individuals, an integrated approach is required to assess which factors, including social rank and bonds to conspecifics, interact to affect an individual's fertility and health, as has been so clearly demonstrated in the work of Jim Henry. PMID- 9401613 TI - The social environment, behaviour and stress--a case study in guinea pigs. AB - In this article we summarize our work on the relationships between the social environment, behaviour and stress in guinea pigs. We confirm what Jim Henry predicted to be a general rule for all mammals including man 20 years ago: The individual's degree of social stress is related to the stability of the pre- and postnatal social environment in which it lives, to the amount of social support which it receives from bonding partners and to the social experiences which it has made during behavioural development. PMID- 9401614 TI - Individual responses to acute and chronic stress in pigs. AB - Pigs can be characterised as resistant (R) or non-resistant (NR) at an early age (1 to 2 weeks) by means of a backtest. In the test the animal is put on its back and the number of bouts of resistance is used to characterise the animal. The test is performed twice with 1 week interval and only pigs that show a consistent response in both tests are classified as either R or NR pigs. On average eighty percent of a population can be classified by this test. R and NR pigs show consistent behavioural, physiological and immunological differences when tested in various challenge test in later life. The R pigs are more sympathetically dominated and showing an active coping style (fight/flight) as described in rats and mice. The NR pigs are more para-sympathetically dominated, resembling the passive coping style (conservation/withdrawal). In intensive husbandry, breeding sows are housed individually and often tethered. After long term tethering these sows show signs of chronic stress; overreaction of the sympathetic nervous system, hypercortisolaemia and disturbed behaviour. The most common disturbed behaviour found in tethered sows is stereotyped behaviour. Most sows develop stereotypies within 1 month after first tethering. Again great differences are found in the amount of stereotypies shown between sows. Some sows spent up to 80% of their active time on this behaviour while others hardly develop stereotypies. Sows showing high levels of stereotypies manage to counteract the sympathetic overreaction caused by the chronic stress of tethering as was shown by a decrease in heart rate during bouts of stereotyped behaviour. In this view stereotypies help the animal to cope with the averse situation of tethering. However, after 8 months of tethering stereotypies are no longer effectively attenuating heart rate. The effect of stereotypies is limited to the initial phase of chronic stress when the animal is striving to regain control. When chronic stress persists stereotypies get dissociated from their effect on the sympatho-adreno medullary system and the animal loses control. PMID- 9401615 TI - Physiological and neuroendocrine correlates of social position in normotensive and hypertensive rat colonies. AB - The hypothesis to be tested was that socially dominant (D) males in a mixed gender rat colony will have: higher blood pressure (BP) decreased hypothalamic pituitary axis (HPA) activity measured by plasma corticosterone (C) and increased sympathetic activity measured by plasma noradrenaline (NA) as compared to socially subordinate (S) males. BP was measured continuously by implanted aortic telemetry (Data Sciences, MN), plasma noradrenaline measured by HPLC with electrochemical detection and plasma C by RIA. Colonies were established using 8 of each sex of 4 strains: SHR, WKY, and our two Y chromosome congenic strains, SHR/y and SHR/a. Social ranking was determined by physical scarring scores, overall locomotor activity and patrol behaviour. D males had higher BP (active dark cycle) across strains compared to subordinates S: SHR-180 vs 148 mmHg, WKY 145 vs 142 mmHg, SHR/y-185 vs 145 mmHg, SHR/a-180 vs 160 mmHg. Using an acute stressor, BP responsiveness was higher in D than in S SHR and SHR/y males. D males had higher NA levels than S males across strains: (SHR-76% increase, WKY 31% increase, SHR/y-29% increase, SHR/a-40% increase). S males had significantly higher C levels than D males across strains (SHR-29% increase, WKY-123% increase, SHR/y-25% increase, SHR/a-61% increase). The hypertensive Y chromosome (SHR or SHR/y) produced higher SBP during the active dark cycle in D males than in D males with a normotensive Y chromosome (WKY or SHR/a). In conclusion, D males from related hypertensive strains showed elevated sympathetic activity measured by plasma NA and reduced HPA activity measured by plasma C as compared to S males. (Supported by HL-48072-04). PMID- 9401616 TI - Using ethological principles to study psychosocial influences on coronary atherosclerosis in monkeys. AB - Studies with male cynomolgus monkeys suggest that atherosclerosis is potentiated among individuals that are habitually successful in their aggressive encounters with social strangers, thereby retaining dominant social status in an unstable environment. Further, the increased risk of atherosclerosis experienced by such animals is related, in part, to the autonomic (sympathetic) adjustments they make while responding to the demands of retaining dominant status. These data provide clear support for the hypothesis that psychosocial factors influence disease pathogenesis via neuroendocrine mediation. Additionally, they provide initial evidence in favor of Jim Henry's suggestion that the pattern of neuroendocrine response to environmental challenge depends on the type and degree of control an animal can exert in such circumstances (Henry & Stephens 1977). PMID- 9401617 TI - The role of psychosocial factors in human disease: lessons from animal models. AB - Studies with humans have identified certain psychosocial characteristics that are associated with increased risk of developing life-threatening illnesses, as well as biological and behavioral mechanisms whereby risk is mediated. In this paper I review research wherein the use of animal models increases our understanding of the mechanisms acting in humans, using research on psychosocial risk factors and the "SES gradient" as examples. PMID- 9401618 TI - A biopsychosocial approach to etiology and pathogenesis. AB - The interactive etiological and pathogenic processes between physical and psychosocial environ-mental stimuli, the individual's appraisal of these influences, and his or her reactions to them in terms of emotion, cognition, behaviour, and physiology; the modification of these reactions through coping, social support, and other interacting variables; and the resulting changes in health and well-being--are extremely complex and poorly understood. Against this background, this paper argues for a biopsychosocial approach, based on an ecological model influenced by, and influencing, James P. Henry's related approaches. This approach is exemplified in six studies carried out by our group and briefly reviewed in this paper. PMID- 9401619 TI - Fighting for and losing or gaining control in life. AB - In JP Henry's work, fighting for and losing control were important concepts in the interpretation of energy mobilization in psychosocial conditions. Attachment and support were important protective and salutogenic factors. These concepts have been applied in a series of epidemiological and psychophysiological real life studies. Job conditions which force the worker to mobilize energy and concomitantly inhibit anabolism could be identified at least partly by means of the demand-control-support model originally proposed by Karasek. The most adverse conditions at work arise when psychological demands are high and at the same time the decision latitude is low. This combination is associated with changes in the regulation of endocrine parameters as well as with increased morbidity--heart disease, functional gastrointestinal symptoms and musculoskeletal disorders. Examples of studies of physiological correlates of psychosocial processes leading to fight for control are also described from outside work activities. PMID- 9401620 TI - Linking sociological with physiological data: the model of effort-reward imbalance at work. AB - While socio-epidemiologic studies documented impressive associations of indicators of chronic psychosocial stress with cardiovascular (c.v.) disease evidence on patho-physiologic processes is still limited. In this regard, the concept of heightened c.v. and hormonal reactivity (RE) to mental stress was proposed and explored. While this concept is a static one we suggest a more dynamic two-stage model of RE where recurrent high responsiveness (stage 1) in the long run results in attenuated, reduced maximal RE due to functional adaptation (stage 2). We present results of an indirect test of this hypothesis in a group of 68 healthy middle-aged men undergoing a modified Stroop Test: in men suffering from high chronic work stress in terms of effort-reward imbalance significantly reduced RE in heart rate, adrenaline and cortisol was found after adjusting for relevant confounders. In conclusion, results underscore the potential of linking sociological with physiological data in stress research. PMID- 9401621 TI - The sympathetic nervous system in human hypertension. AB - While animal models of hypertension have clearly shown an increase in sympathetic activity, a similar demonstration in humans has been more difficult to obtain for methodological reasons. There is now clear evidence, however, of an increase sympathetic activity in essential hypertension by the finding of either an increase in plasma norepinephrine and an increase in muscle sympathetic nerve traffic. Among the mechanisms responsible for this sympathetic activation the arterial baroreflex may play a role although probably a non specific and late one. Central neural influences associated with an excessive hypothalamic response to stress may also be involved, but conclusive evidence is still lacking due to the difficulty of assessing cardiovascular reactivity to stress in man. A deeper insight into the features of human sympathetic cardiovascular control may be offered now by new techniques which allow neural cardiovascular regulation to be assessed in daily life through computer analysis of noninvasive ambulatory beat by-beat blood pressure and heart rate recordings. PMID- 9401622 TI - Continuing on J.P. Henry's path; studies of physiology and pathophysiology of cardiopulmonary receptors in humans. AB - The seminal work of Henry and Gauer lead us to a) show that in healthy humans cardiopulmonary receptors (CPR) regulate renin release, b) that low renin hypertension is associated with an expanded cardiopulmonary volume. This suggested that the low renin state in hypertension may be due to excessive inhibition of renin by the CPR. In the course of these experiments we uncovered an undescribed pressor reflex which was used to investigate the effect of intermittent pressor episodes on cardiac structure in dogs. Finally, in recent years, we used unloading of CPR to show that reflex vasoconstriction causes acute insulin resistance in the human forearm. PMID- 9401623 TI - The effects of occupational stress on blood pressure in men and women. AB - Human hypertension is the end result of a number of genetic and environmental influences, and typically develops gradually over many years. The sympathetic nervous system appears to play a role in the early stages, with structural changes in the resistance vessels becoming dominant later on. The extent to which increased sympathetic actively may be the result of environmental stress is uncertain. Animal studies have suggested that chronic stress can raise blood pressure. Human epidemiological studies have shown that the prevalence of hypertension is strongly dependent on social and cultural factors. Blood pressure tends to be highest at work, and studies using ambulatory monitoring have shown that occupational stress, measured as job strain, can raise blood pressure in men, but not women. This may be associated with increased left ventricular mass. The diurnal blood pressure pattern in men with high strain jobs shows a persistent elevation throughout the day and night, which is consistent with the hypothesis that job strain is a causal factor in the development of human hypertension. PMID- 9401624 TI - Stress hypertension: the "wrong" genes in the "wrong" environment. AB - Jim Henry demonstrated an animal's society can induce an increase in blood pressure and its cardiovascular sequale. He recognized that the stress required to elevate blood pressure was a function of the genetically determined behavioral traits of the mice used. He termed some strains aggressive, others peaceable. Being highly inbred (indeed isogenic strains) it was intriguing to find that the behavior of these genetically identical individuals could differ markedly once placed in a society that decreased territory. A dominant or "king" mouse emerged. Other non-dominant males were aggressive and striving to be king. Adrenal medullary systems were activated and renins high. Others huddled in one cage and appeared to have given up. Jim called them depressed. Their adrenal cortex was hyperplastic suggesting pituitary adrenal axis activation as in depression, their renin was low and corticosterone high. In rats, careful selection of a strain genetically aggressive had to be combined with titration of societal stress to reliably induce hypertension. Its likely that humans retain some, if not all, of these variations, i.e. some respond to stress with an increase in blood pressure and others do not, some respond via the sympathetic pathway and others by adrenal cortical activation. The suggestion that African American's high blood pressures is due to stress is relevant to the Henry paradigm and the known genetic influences on sodium retention in blacks. The integration of this paradigm with the genetically increased sensitivity to the blood pressure raising effects of dietary sodium in blacks is proposed and discussed. PMID- 9401625 TI - Do environmental effects on human emotions cause cardiovascular disorders? AB - Environmental influences on human health include the effects of toxic materials and adverse ecological factors. Natural milieu stressors also affect emotions that may adversely affect cardiovascular function and precipitate or otherwise contribute to complications of cardiovascular diseases. However, although variously hypothesized, there is inadequate evidence that they directly contribute to the pathogenesis of sustained hypertension or coronary atherosclerosis. PMID- 9401626 TI - Blood pressure trend and psychosocial factors: the case of the nuns in a secluded order. AB - The powerful effect of psychosocial and acculturating influences on population blood pressure trends seems to be confirmed, through longitudinal observations in the nuns in a secluded order. After initial observation had been made on culture, body form, blood pressure, diet, and other variables in 144 nuns and 138 lay women, included as a control group, a 30-year follow-up study was undertaken. Most striking were opposite trends noted between the two groups in blood pressure trend. During the follow-up period blood pressure remained remarkably stable among the nuns. None showed a rise in diastolic blood pressure to above 90 mmHg. By contrast, the control women showed the expected increase in blood pressure with age. This resulted in a gradually greater difference (A > 30/15 mmHg) in systolic and diastolic blood pressure between the two groups, which was statistically significant. It appears reasonable to attribute much of the difference in blood pressure to the different burden in psychosocial factor and to the preserved peaceful lifestyle of the nuns. PMID- 9401627 TI - Visceral responses to the social environment. AB - Twenty-five hundred years ago, Hippocrates, recognizing the impact of life experience on health wrote: "those things which one has been accustomed to for a long time, although worse than things to which one is not accustomed, usually give less disturbance" (Bruhn & Wolf 1978). Since Hippocrates, much evidence has emerged to suggest that social stability is conducive to health, while social change, especially rapid change, may predispose to illness. The idea that sensory information from ordinary life experiences contributes in a major way to shaping the activities of the brain, took root in the 18th century with Pierre Gassendi and John Locke. A mechanism whereby the vast numbers of neurons in the brain could be rapidly rearranged and organized to respond to a host of different external stimuli emerged from the work of Santiago Ramon y Cajal who discovered the axonal growth cone and its capacity to elongate and guide an axon a considerable distance to a target dendrite (Cajal 1890). Thereby, with very rapid shifts in attention and perception, a wide range of responses, emotional, and physiological can be elicited. With this and other evidence at hand that the intracranial regulatory mechanisms can reorganize in response to environmental change including social change, we undertook a 30 year (1962-1992) prospective study of the effects of social patterns on health in Roseto, a closely knit Italian-American incorporated town in Pennsylvania. It was settled in 1882 by Italians from Roseto val Fortore, a town in southern Italy near the Adriatic Sea. We found a sharp increase in myocardial infarction among Rosetans when long established cohesive social patterns began to weaken and be replaced by more egocentric attitudes. PMID- 9401628 TI - Stress and cardiovascular disease. AB - The statistical associations between stress and cardiovascular and other prevalent diseases have not been explained. Perceived stress, resulting in an uncontrollable defeat reaction, has been shown by James Henry (Henry 1993) to be followed by specific endocrine abnormalities, including sensitization of the hypothalamo-pituitary-adrenal (HPA) axis, and inhibited sex steroid and growth hormone secretions. With an elevated waist/hip circumference ratio (WHR)--a simple, surrogate, measurement of intraabdominal, visceral fat masses--combined with insulin resistance, similar endocrine perturbations are found. Based on considerable evidence, such endocrine abnormalities seem to be followed by accumulation of intraabdominal, visceral fat masses and insulin resistance, both powerful risk factors for cardiovascular disease, diabetes and stroke. A postulated chain of events is therefore that the endocrine perturbations are primary factors, followed by visceral fat accumulation, insulin resistance and other risk factors dependent on the hyperinsulinemia following insulin resistance. This highlights the importance of elucidating the cause(s) to the endocrine abnormalities. These are identical to those described by Henry (1993) to follow a stress reaction of a defeat type. Findings of several psychosocial and socio-economic handicaps might provide a basis for such a reaction, supported by experimental studies in primates. Furthermore, depression, anxiety, alcohol consumption and smoking, all known activators of the HPA axis, are also often found. The low sex steroid and growth hormone secretions might be secondary to the hypersensitive HPA-axis. They could also be caused by other factors, and are, each alone, capable of causing both visceral fat accumulation and insulin resistance. Visceral fat accumulation is only an indirect, surrogate measurement of the underlying endocrine abnormalities, but is useful for screening purposes on a population basis. Developments of novel techniques for sensitive, yet simple measurements of HPA axis activity under undisturbed conditions seem to allow a better definition of pathogenetic factors. Utilizing such methods, subgroups of the syndrome including visceral fat accumulation, insulin resistance and other associated risk factors (Metabolic Syndrome), are beginning to emerge. A more detailed information on noxious factors in society leading to a defeat reaction to perceived stress, endocrine abnormalities and the Metabolic Syndrome, with increased risk for prevalent disease may hopefully be developed by these new methods. PMID- 9401629 TI - Heart disease mortality and economic changes; including unemployment; in Western Germany 1951-1989. AB - Research over the last two decades has indicated that changes in cardiovascular disease mortality rates have been influenced by those in national economic indicators as well as by measures of consumption of tobacco, animal fats and alcohol. These findings predominantly involved the United States, United Kingdom and Scandinavian countries. The economic indicators included real per capita income and social welfare expenditures (beneficial relationships to mortality rates), and unemployment rates and business failure rates (detrimental relationships to mortality). James Henry's formulations have emphasized that many different illnesses respond to emotional stresses in different psychophysiological patterns depending on the specific constellations of emotions aroused. On the assumption that the impact of national economic changes on cardiovascular mortality reflects emotional stresses, losses, frustrations and deprivations, similar tests were undertaken using Western German heart disease mortality rate data over 1951-1989. Time-series regression analysis showed that, holding constant the effects of tobacco, animal fats and alcohol, increased income and social welfare expenditures are related to heart disease mortality rate declines, whereas increased unemployment and business failure rates are related to heart disease mortality rate increases over more than a decade. PMID- 9401630 TI - Exhausted subjects, exhausted systems. AB - A state of 'vital exhaustion', characterized by unusual tiredness, increased irritability and feelings of demoralization has been found to preceed the onset of myocardial infarction and to increase the risk of a new coronary event after angioplasty. Probably this state reflects a decreased activity of the hypothalamic-pituitary-adrenal axis as part of an homeostatic reaction to prolonged stress and inflammation. PMID- 9401631 TI - Stress management & hypertension. AB - Review of literature suggests that emotional and social stress is a contributory factor to the development of hypertension. If so, relaxation and stress management therapy may reduce high blood pressure (BP) and its complications. In a series of studies I and my colleagues have shown that this may be so. These studies have been published elsewhere and are briefly summarized here. PMID- 9401632 TI - Changes in cardiovascular risk factors and hormones during a comprehensive residential three month kriya yoga training and vegetarian nutrition. AB - In participants of a comprehensive residential three month yoga and mediation training programme living on a low fat lacto-vegetarian diet changes in cardiovascular risk factors and hormones were studied. Substantial risk factor reduction was found. Body mass index, total serum and LDL cholesterol, fibrinogen, and blood pressure were significantly reduced especially in those with elevated levels. Urinary excretion of adrenaline, noradrenaline, dopamine, aldosterone, as well as serum testosterone and luteinizing hormone levels were reduced, while cortisol excretion increased significantly. PMID- 9401633 TI - The consequences of having experienced overwhelming stress. PMID- 9401634 TI - Traumatic stress and attachment. AB - Traumatic stress in the normal individual results in activation of the sympatho adrenal system causing a rise in noradrenaline and adrenaline, stimulation of the thyroid system causing increased secretion of thyroid hormones and activation of the hypothalamic-pituitary-adrenocortical (HPA) system resulting in elevated levels of cortisol. Studies in animals and in humans with posttraumatic stress disorder indicate that chronic traumatic stress can result in dissociation of the sympatho-adrenal medullary and the HPA system resulting in sustained elevations of the former system but suppressed or altered ACTH-corticoid responsivity. As reviewed by Henry, self preservative behavior with its emphasis on power and control, is associated with catecholamines, thyroid hormones and left hemispheric functioning while species preservative behavior, with its emphasis on attachment, familiarity, reverence and synchronicity, is associated with cortisol, oxytocin and right hemispheric functioning. Traumatic stress seems to disturb this hemispheric balance which is reflected in the suppression of cortisol and loss of attachment behavior and other species preservative right hemispheric functions. PMID- 9401635 TI - James P. Henry. Revisited in reference to his Instincts, Archetypes, and Symbols: an approach to the physiology of religious experience. AB - In his Instincts, Archetypes, and Symbols James P. Henry places physiological aspects of religious experience in the context of cultural anthropology and religious philosophy. His approach is Jungian. Of major concern is the liberation of the human mind from privatizations of goodness (privatio boni). Henry characterizes optimal natural conditions for a healthy psyche substantiated by physiological research. PMID- 9401636 TI - James Paget Henry: a man for all seasons. AB - Jim Henry is best known for his seminal research on psychosocial stress and cardiovascular disease, and studies of the neuroendocrine correlates of animal behaviors during stress. However, he had a wide range of interests, made important contributions in many other areas, and served as a remarkable catalyst in advancing the work of other scientists. These numerous and less appreciated accomplishments could not possibly be covered in an article of this length. Hopefully, some personal observations, and a brief sketch of his extension of Carl Jung's research and his involvement in the U.S. space program, may help to illustrate the superb character and qualities of this modest and multifaceted individual, as well as a few of his lesser known, but equally meaningful achievements. PMID- 9401637 TI - Jim Henry's world revisited--environmental "stress" at the psychophysiological and the molecular levels. AB - Ever increasing applications of sophisticated technologies in western civilization have placed great and growing demands for the rapid and accurate processing of multi-modal sensory information. These information streams may exceed an individual's performance capabilities. Failure to respond appropriately may have serious consequences, not only for the individual but also for others, as in command situations in the aerospace environment. There are, for example, consistent patterns common to EEG records in a population of astronaut candidates, when exposed to increasing visual information overload, simulating hazardous flight conditions. The records are dominated at the point of "information overload" by sharply and progressively increased theta wave (4-7 Hz) activity in temporal regions, major increments in frontal beta (> 14 Hz) activity, and markedly reduced occipital alpha (8-12 Hz) levels. These responses to a simulated stress raise questions about the brain's ability to distinguish natural reality from the mediated reality in modern life. It has been hypothesized that an individual's reactions with computers, television and new media are fundamentally social and natural, just as in interactions in real life. Also immune responses may here offer valuable benchmarks concerning reactions to mentally stressful stimuli. Another type of environmental influences in modern society is that of electromagnetic fields. Even fairly weak (athermal) electromagnetic fields have proven to be useful tools to study regulatory mechanisms in cells from brain and other tissues. There is growing evidence that nitric oxide may influence normal EEG patterns and that it may also participate in the pathophysiology of oxidative stress disturbances, including influences in e.g. Parkinson's and Alzheimer's diseases, then behaving as a free radical with reactive-oxygen-species or reactive-nitrogen-species. As a free radical, nitric oxide is sensitive to a variety of imposed magnetic fields, with theoretical and experimental evidence that its actions in regulating the rate and amount of product of cerebral biochemical reactions may also be modulated by imposed magnetic fields. PMID- 9401638 TI - Transcription factor abnormalities as a cause of beta cell dysfunction in diabetes: a hypothesis. AB - Well-characterized defects in insulin secretion, most notably a loss of glucose induced insulin secretion, are found in virtually all forms of NIDDM, as well as in early IDDM. Similar abnormalities have been found in all animal models of diabetes in which they have been studied. A novel hypothesis is being proposed to explain the mechanisms responsible for these alterations. Many abnormalities in the various steps of glucose-induced insulin secretion have been identified in rodent models of diabetes, but none by itself seems sufficient to explain the defects. These include a loss of GLUT2, glycogen accumulation, glucose recycling, abnormal glucokinase or hexokinase, altered mitochondrial glycerol phosphate dehydrogenase (mGPDH) activity, abnormal ion channel function and beta cell degranulation. We propose that optimal secretory function is dependent upon the unique differentiation of beta cells that is maintained by a set of transcription factors and that this control is disrupted by the diabetic state. Therefore, we propose that key transcription factors are affected even when beta cells are stressed by insulin resistance in very earliest stages of diabetes and that the abnormality becomes more severe as full-blown diabetes develops, which leads to loss of beta cell differentiation and a resultant derangement of insulin secretion. PMID- 9401639 TI - Islet autoantibodies and their use in predicting insulin-dependent diabetes. PMID- 9401641 TI - Prevalence and 4-year incidence of insulin-dependent diabetes mellitus in the province of Oristano (Sardinia, Italy). AB - The aims of this study were: (1) to estimate the prevalence of insulin-dependent diabetes mellitus (type I) among the population of 0-29-years-old residents in the province of Oristano on 31/12/96; (2) to estimate the incidence of type I diabetes among the population of 0-29-years-old residents in the province of Oristano between 1993 and 1996. Data concerning the cases were collected from two independent sources. The capture-recapture method was utilised to estimate ascertainment probability. Demographic data for the resident population were supplied by 78 communes in the province. The completeness of the primary data source for prevalence was 97.1%; the estimated prevalence was 4.61 per thousand. The number of type I diabetic subjects identified in a population of 60,095 people aged 0-29 years was 276, i.e. a prevalence of 4.59 per thousand (95% confidence intervals [CI] 4.07-5.17). No cases were found in the commune of Arborea, where a large percentage (65%) of immigrants from the Veneto region live (total population < 30 years of age = 1706; 8 expected cases; P < 0.001). The M/F ratio was 1.12. The completeness of primary source of incidence data was 100%. Between 1993 and 1996, the number of new type I diabetic subjects was 86, with a M/F ratio of 1.00. The age-standardised incidence rates were: 54.4 (95% CI 41.3 70.1) in the 0-14-year-age group, 22.0 (95% CI 15.2-30.5) in the 15-29-year-age group and 38.2 (95% CI 31.0-46.4) in the 0-29 year age group, per 100,000 person years. The incidence and prevalence of type I diabetes mellitus are very high, ranking the province of Oristano among the highest in Europe. PMID- 9401640 TI - Accelerated gastric emptying during hypoglycaemia is not associated with changes in plasma motilin levels. AB - This study examined whether or not changes in plasma concentrations of motilin and other gastrointestinal hormones known to affect gastric motility are associated with the accelerated gastric emptying seen during hypoglycaemia. While studying gastric emptying by scintigraphy in eight healthy subjects, the plasma concentrations of glucagon, adrenaline, motilin, gastrin, neuropeptide Y and somatostatin were measured during normoglycaemia and hypoglycaemia with simultaneous infusion of either atropine or saline. Blood glucose concentrations were checked by an insulin-glucose clamp. The plasma levels of glucagon and adrenaline increased markedly during both hypoglycaemic examinations compared with normoglycaemia. Neither motilin nor any of the other hormones displayed considerable changes during hypoglycaemia with and without atropine compared with normoglycaemia. No further information about the mechanisms behind the accelerated gastric emptying rate during hypoglycaemia was obtained by analysing motilin and the other gastrointestinal hormones. PMID- 9401642 TI - Acute effects of pioglitazone on glucose metabolism in perfused rat liver. AB - Pioglitazone, a thiazolidinedone derivative, decreases insulin resistance and improves hyperglycemia in insulin-resistant obese and/or diabetic animals. However, the mechanisms by which hyperglycemia is improved are not well defined. We investigated the effects of pioglitazone on hepatic glucose metabolism using a perfused rat liver model. Perfusion with the buffer containing 1-10 microM pioglitazone for 20 min dose-dependently increased the hepatic fructose 2,6 bisphosphate content, a potent activator of 6-phosphofructo 1-kinase. The fructose 2,6-bisphosphate level after 20 min perfusion with 10 microM pioglitazone was 64.9 +/- 14.5 pmol/mg.protein, significantly higher than the control (48.3 +/- 10.9 pmol/mg.protein). When the liver from a starved for 48 h rat was perfused with the buffer containing 2 mM lactate but no glucose, glucose was generated from lactate via the gluconeogenic pathway and flowed into the effluent perfusate at a constant rate of 31 +/- 0.6 mumol/g.liver/h. The addition of 10 microM pioglitazone decreased the glucose output rate to 19.3 +/- 3.8 mumol/g.liver/h. Dose-dependent inhibition of glucose output by pioglitazone was observed in the 1-10 microM dose range. These results indicate that pioglitazone may not only stimulate glycolysis but also inhibit gluconeogenesis in the liver. These acute and insulin-independent effects on hepatic glucose metabolism may partly account for the diverse anti-diabetic effects of pioglitazone. PMID- 9401643 TI - Relative weight of glucose, insulin and parathyroid hormone in the urinary loss of phosphate by chronically diabetic rats. AB - This report deals with the relationships between glucose (G) and insulin on the tubular transport of phosphate (P) in chronically diabetic rats with high plasma levels of parathyroid hormone (PTH). Alloxan-induced diabetes leads to phosphorus depletion of the soft tissues. This phenomenon appears associated with weight loss and negative P balances caused by the increased urinary P excretion. Administration of 2 IU of insulin/100 g body weight (bw) to diabetic rats normalized their P balance and body weight. The effect of parathyroid function on the P metabolism of diabetic rats was investigated with balance experiments. Diabetic rats, intact or thyroparathyroidectomized (TPTX), have a greater urinary excretion of P than their controls. However, in control rats, the ratio intact:TPTX for urinary P is 1.0:0.76, showing the antiphosphaturic effect of parathyroid ablation. For diabetic animals, on the other hand, the ratio is 1.0:1.44. The simultaneous deficit of insulin and PTH thus quadruples the urinary P loss, instead of compensating for each other. The contribution of insulin deficit and hyperglycemia to the defect in tubular reabsorption (TRP) was investigated with clearance experiments (done on anesthetized, perfused rats). Five experimental groups were used: Controls (C), diabetics (D), controls + glucose (C + G), diabetics + insulin (D + I) and diabetics + insulin + glucose (D + I + G). All experimental groups showed a linear relationship between the TRP of P and G. The regression equation for C is significantly different (F = 40.1, P < 0.001) from that of D animals. The slope value measure the number of mumoles of P per mumol of G reabsorbed. For C and D rats, the ratio P:G approximates 1:4 and 1:20, respectively. The increase in P:G ratios represents the competition between both substrates for tubular resorption. Glycemias up to 11 mM (C and D + I) exist concurrent with the P:G ratio 1:4 Glycemias above 25 mM (D, C + G and D + I + G) produce a P:G ratio of 1:20. Fractional excretion of P (FEP) increased significantly in untreated, chronically diabetic rats (0.47 +/- 0.12 vs controls = 0.05 +/- 0.01, P < 0.001). After a single intramuscular injection of insulin, the FEP decreased as a function of insulin levels. To normalize the FEP of diabetic rats in short-term experiments, insulin had to be administered in doses that produce plasma insulin levels 25 times greater than normal. The general information afforded by the present experiments shows that in untreated, chronically diabetic rats, insulin deficit plays an indirect role. The absence of PTH enhances the effect of hyperglycemia. The latter and the concurrent tubular overload of glucose are the cause of hyperphosphaturia in these animals. PMID- 9401644 TI - Glycosylated hemoglobin as predictor of adverse fetal outcome in type 1 diabetic pregnancies. AB - The study aimed to determine whether a consistent dose-response association can be demonstrated, after adjustment for maternal age and White classification, between glycosylated hemoglobin (HbA1c) values before conception and in the first trimester of pregnancy of insulin-dependent diabetic mothers and adverse fetal outcome (abortions and major malformations). This is a historical follow-up study based on medical records in a geographically defined catchment area. The study comprised 60 pregnancies with HbA1c determinations before pregnancy and 161 with HbA1c in the first trimester in women with type 1 diabetes admitted between 1980 and 1992. Relative risk calculations indicated a highly significant and consistent correlation between HbA1c values above 6.6% and adverse fetal outcome after adjustment for differences in maternal age and White classification. Our data support a clinically significant and consistent relationship between adverse fetal outcome and HbA1c in the first trimester of pregnancy of type 1 mothers, without any indication of a cut-off level below which further improvement in HbA1c was of minor importance. PMID- 9401645 TI - Erythrocyte Na+/H+ exchange and preclinical abnormalities of the left ventricular diastolic function in normotensive type 1 (insulin-dependent) diabetic patients. AB - We assessed the relationship between erythrocyte Na+/H+ antiport activity and myocardial anatomical-functional parameters (by Doppler echocardiography) in normotensive IDDM patients, with and without microalbuminuria. We studied 33 normotensive IDDM subjects and 14 matched healthy controls (group 4). Based on urinary albumin excretion rate (UAER), 23 diabetics were normoalbuminuric, 10 microalbuminuric (group 3). Normoalbuminurics were divided up for normal (group 1, n = 13) or high (group 2, n = 10) antiport activity. We evaluated fasting glycaemia and 24-h urine glucose output, HbA1c, plasma lipids, urea, creatinine and electrolyte clearances, UAER, erythrocyte Na+/H+ countertransport, M-Mode and 2D echocardiograms with Doppler analysis. Antiport, which was higher in diabetics than controls, was significantly overactive in groups 2 and 3 vs group 4, independently from UAER. Diabetics showed left ventricular volume, cardiac mass and systolic function within the control range. In left ventricular diastolic filling, while peak E was similar in diabetic and healthy people, the late peak transmitral flow velocity (peak A) was significantly higher in diabetics than controls, and this was also true in groups 2 and 3 vs group 4. Antiport activity was positively related to peak A (p < 0.03). These observations suggest that (a) the Na+/H+ antiport may be overactive in diabetes, apart from microalbuminuria; (b) increased Na+/H+ antiport activity, in normotensive IDDM people, may be associated with preclinical diastolic myocardial dysfunction ("incipient diabetic cardiomyopathy"?). PMID- 9401646 TI - A liquid mixed meal or exogenous glucagon-like peptide 1 (GLP-1) do not alter plasma leptin concentrations in healthy volunteers. AB - Glucagon-like peptide 1 [7-36 amide] (GLP-1) and the obese gene product (leptin) are thought to be involved in the central regulation of feeding. Both may act from the peripheral circulation to influence brain function. To study potential interactions, GLP-1 ([7-36 amide]: 0.4, 0.8 pmol kg-1 min-1 or placebo on separate occasions) was infused intravenously (from -30 to 240 min) into nine healthy volunteers [age 26 +/- 3 years, body mass index: 22.9 +/- 1.6 kg/m2, glycated haemoglobin HbA1c: 5.0% +/- 0.2% (normal: 4.0%-6.2%), creatinine: 1.1 +/ 0.1 mg/dl], and (at 0 min) a liquid test meal (50 g sucrose in 400 ml 8% amino acid, total amino acids 80 g/l) was administered via a nasogastric tube. Plasma leptin (radioimmunoassay, RIA), glucose, insulin (microparticle enzyme immunoassay), C-peptide (enzyme-linked immunosorbent assay) and GLP-1 (RIA) were measured, and statistical analysis was done with repeated-measures ANOVA and Student's t-test. Plasma leptin concentrations were 31 +/- 6 pmol/l in the basal state. They did not change within 240 min after meal ingestion nor in response to the infusion of exogenous GLP-1 [7-36 amide] (P = 0.99 for the interaction of experiment and time) leading to GLP-1 mean plasma levels of 25 +/- 2 and 36 +/- 3 (basal 6 +/- 1) pmol/l. On the other hand, glucose (from basal 4.7 +/- 0.1 to 6.0 +/- 0.2 mmol/l at 15 min, P < 0.05) and insulin (from basal 28 +/- 2 to 325 +/- 78 pmol/l at 45 min, P < 0.05) increased clearly after the meal with placebo. In conclusion, (1) plasma leptin levels in normal human subjects show no short-term changes after feeding a liquid mixed meal and (2) do not appear to be directly influenced by physiological and pharmacological elevations in plasma GLP-1 [7-36 amide] concentrations. This does not exclude interactions at the cerebral (hypothalamic) level or on more long-term temporal scales. PMID- 9401647 TI - Free plasma magnesium following glucose loading in healthy humans. AB - The recognized existence of a circadian pattern in extracellular magnesium balance might mirror either an inherent rhythm in the homeostasis of this ion or dietary factors. Since in vitro insulin enhances cellular magnesium uptake, the circadian rhythm in extracellular magnesium metabolism might be modulated at least in part by carbohydrate intake. To assess this hypothesis, the effects of oral glucose loading on plasma total and ionized magnesium were investigated in lean healthy humans with a negative family history for essential hypertension and diabetes mellitus. Plasma total and ionized magnesium was similar before glucose loading and 30, 60, 90, 180, and 210 min thereafter. It is therefore concluded that in healthy humans the circadian pattern of extracellular magnesium is not modulated by the metabolic and hormonal mechanisms that adjust the concentration of glucose. PMID- 9401648 TI - Haematoma evacuation does not improve outcome in spontaneous supratentorial intracerebral haemorrhage: a case-control study. AB - Surgical intervention in supratentorial intracerebral haemorrhage (ICH) is still controversial. We assessed the value of haematoma evacuation with a case-control study. 145 consecutive patients with supratentorial spontaneous ICH without tumour or vascular abnormalities were analysed. Haematoma evacuation was performed in 24 patients. Age, sex, Glasgow Coma Scale (GCS), level of consciousness, pupillary reaction on admission, localisation, aetiology and volume of the haematoma, presence of ventricular blood, and Glasgow Outcome Scale (GOS) on discharge were analysed. From statistical analysis 40 patients > 80 years and with haematoma volume < 10 ml, who were always treated conservatively, were excluded. Prognostic factors retained from a multiple regression model with the dichotomised GOS scale (GOS 1-3, 4 + 5) as response variable were GCS, haematoma volume and location. The only difference between all medically treated and "operated" patients was haematoma volume, which was larger in the "operated" patients. All 24 evacuated cases could be matched to a medically treated control regarding age, haematoma volume and location, GCS, and pupillary reaction. Significant differences between the two groups could not be detected. Outcome was not different between the two groups. After separating the sample into patients with and without ventricular haemorrhage, there was no different outcome between the two groups either. We conclude that haematoma evacuation did not improve outcome in supratentorial spontaneous ICH. Since haematomas were evacuated mainly in clinically deteriorating patients, our data suggest that the only effect of haematoma evacuation is to stop progressive deterioration rather than to improve overall clinical outcome. PMID- 9401649 TI - Treatment results of acromegaly as analyzed by different criteria. AB - Results of treatment of acromegaly are often incomparable due to the different criteria which have been used for defining cure or control of disease. At the present time it is widely accepted, that the main criteria of cure must be normalization of IGF-1 and a GH in the OGTT < 2 ng/ml. In this retrospective study we investigated the endocrinological results of 56 patients, who were surgically treated because of a GH-producing pituitary adenoma, by different criteria. Twelve of our patients had had additional medical treatment after surgery, two received radiotherapy. At a mean follow-up of 34 months after surgery 66% of patients had a basal GH < 5 ng/ml, 64% had a GH in the OGTT < 2 ng/ml and 73% had normalization of IGF-1. The combined criteria of OGTT < 2 ng/ml and IGF-1 normalization have been fulfilled in 59% of patients. None of these latter patients developed a clinical recurrence during the follow-up period. An optimal result (endocrinological cure, no permanent surgical complications and intact pituitary function) was achieved in 43% of patients. Although surgery was responsible for 19 new pituitary axis deficiencies (7 corticotropic axis, 8 thyrotropic axis and gonadotropic axis), 22 partial deficiencies improved to normalization after surgery (respectively 6, 3, and 13). Pre-operatively 55% of patients had no pituitary deficiency, after surgery this was 61%, leaving a net positive result of 6% less pituitary deficiencies. The authors conclude that normalization of IGF-1 combined with an OGTT < 2 ng/ml are adequate criteria for the definition of cure of acromegaly. However, the authors propose to include post-treatment hypopituitarism as an additional criterion by which treatment of acromegaly should be evaluated. PMID- 9401650 TI - Early radiologically proven rebleeding from intracranial cavernous angiomas: report of 6 cases and review of the literature. AB - Although intracranial cavernomas are known to cause haemorrhage, data concerning the frequency, severity and delay of recurrent bleedings are controversial. We report a series of 6 patients with histologically proven cavernoma, presenting with early clinical signs and radiological proof of rebleeding, that is occurring in the first month after initial overt haemorrhage. These 6 cases have been selected from a series of 142 patients seen between 1980 and 1995 in our department with cavernous angiomas or so-called AOVMs, of whom 93 presented with clinical symptoms of haemorrhage (34 patients presented symptoms of one or more rebleeding, but only 6 had radiological proof). All patients suffered neurological worsening due to the rebleeding, with an increase of the size of the haematoma on the CT scan. Five MRIs were performed at the acute stage: 3 showed evidence of cavernoma (60%). All patients underwent surgery at the acute stage of the rebleeding, with 5 improvements and 1 stabilization. A cavernous angioma was found in 5 cases at first surgery, but a further operation was necessary in the last patient to find and remove the cavernoma, after a second rebleeding following the first intervention. Our series reveals a high frequency of rebleeding after a first intracranial haemorrhage from a cavernous angioma, and highlights the precocity of such rebleedings. Therefore, we advocate early aggressive surgical management: in cases of cavernoma revealed by a first clinical overt haemorrhage, when there is strong radiological suspicion at the acute stage; and in all cases of rebleeding, even without radiological evidence of malformation, in the absence of vascular risk factors. Surgical indication must be discussed in particular cases of cavernomas of the brain stem when neither the haematoma nor the cavernoma reach the surface, and in deep supratentorial cavernomas, when the neurological status is good, because of the therapeutic risk. PMID- 9401651 TI - Ruptured vertebrobasilar junction aneurysm associated with basilar artery fenestration. AB - A case of a ruptured saccular aneurysm arising from the proximal portion of a partially duplicated basilar artery in a 36-year-old woman is reported. CT and lumbar puncture confirmed subarachnoid haemorrhage. Cerebral angiography detected a vertebrobasilar junction aneurysm associated with basilar artery fenestration. The patient underwent successful clipping and coating of the aneurysm by a right lateral suboccipital osteoclastic approach. Embryological development, pathogenesis, diagnostic and therapeutic difficulties of this vascular malformation are discussed in this report. PMID- 9401652 TI - Growing skull fractures: a clinical study of 41 patients. AB - Growing skull fractures are rare complications of head injury, occurring almost exclusively in infants and children under the age of three. A retrospective review at our Institute yielded 41 patients with this entity over a period of 20 years (1975-1995). The age at presentation ranged from less than 1 year to 62 years, with 33 (80.5%) patients being less than 5 years of age. The cause of injury was either a fall from a height (93%) or a road traffic accident. The most common location of a growing skull fracture was either parietal or frontoparietal (56%). One patient had a posterior fossa growing skull fracture. CT scan was performed in 19 patients which demonstrated an underlying porencephalic cyst, hydrocephalus or a cyst communicating with the ventricle. In 5 children, a ventriculo-peritoneal shunt alone was performed. Twenty four patients underwent a duro- and cranioplasty while a duroplasty alone was performed in 8 patients. The material used for cranioplasty included acrylic, wire mesh, steel plates or autologous bone. Three patients died, one due to an anaesthetic complication and two as a result of postoperative meningitis. Post-operative CSF leaks occurred in 3 patients, which were managed by a lumbar drain. Six patients had local wound infection. PMID- 9401653 TI - Idiopathic normal pressure hydrocephalus: analysis of factors related to cerebrospinal fluid dynamics determining functional prognosis. AB - This investigation has been undertaken to analyze the findings with both the cerebrospinal fluid (CSF) pressure (Pcsf) and CSF pulse pressure (PP) in order to predict the outcome of patients with the syndrome of idiopathic normal pressure hydrocephalus (NPH). Accordingly, a prospective clinical study was planned in which two groups of patients with NPH, having analogous prevalence of several matched clinical and radiological parameters, were separated on the basis of their positive or negative response to shunting. Both the resting Pcsf and CSF PP profiles were compared in these two groups, and between them and normal controls. CSF PP amplitude and CSF PP latency correlated directly in conditions associated with either normal or high compliance (controls and patients with Alzheimer-like disorders), whereas this correlation was inverse in states of low compliance (NPH). On the other hand, shunt-responders showed a resting Pcsf significantly higher than both non-responders and controls. The following conclusions were obtained: 1) CSF PP is a high-amplitude and relative low-latency wave in NPH when compared with controls: 2) CSF PP amplitude and latency correlate directly in normal subjects and in those with primary cerebral atrophy; 3) a non-reversible stage of NPH could be conceived in contradistinction to the reversible one, in both of which an inverse correlation between the amplitude and the latency takes place, the main difference between them being the resting Pcsf, which is significantly lower in the former than in the latter, depending on the degree of atrophic changes developed. PMID- 9401654 TI - Costs and effects of microsurgery versus radiosurgery in treating acoustic neuroma. AB - This study analyses costs and effects of treating acoustic neuroma patients by using microsurgery compared to radiosurgery. Radiosurgery is the stereotactic application of radiotherapy and an innovative medical technology. Cost and effect estimates of conventional treatment were based on a retrospective study in the Netherlands. Similar data for a comparable group of patients in Sweden were collected for radiosurgery, as this treatment option is currently not available in the Netherlands. Fifty-three acoustic neuroma patients who had been operated on the University Hospital Rotterdam between November 1990 and February 1995 were included. This group was compared with 92 acoustic neuroma patients treated with radiosurgery (Gamma Knife. Stockholm, Sweden) in the same period. Data on health care use were collected from patient files. To obtain data on production losses and quality of life, a questionnaire was sent by mail in February 1995. This booklet consisted of the Health and Labour-questionnaire (HLQ), the Short Form-36 (SF36) and the EuroQol. The response rate was 92%. Direct costs for microsurgery amounted to Dfl. 20.072,- and for radiosurgery to Dfl. 14.272,-. Indirect costs were respectively Dfl. 16.400,- and Dfl. 1.020,-. General health rating was better for radiosurgery than for microsurgery. On the whole, differences in clinical outcomes between the two patient groups were small. Assuming a reasonable occupancy rate of the expensive radiosurgery equipment, we demonstrated that for the short term treating patients with acoustic neuroma with an extra-meatal tumour diameter of less than 3 centimeters, radiosurgery is more cost-effective than microsurgery. PMID- 9401655 TI - Peripheral nerve schwannomas--an analysis of 16 patients. AB - 16 patients with peripheral nerve neurinomas (benign schwannomas) were operated upon in our hospital between 1990-1995. The largest tumours were found on proximal segments of peripheral nerves (brachial plexus: 15 cm, sciatic nerve: 20 cm). The average duration of symptoms was 1 1/2 years (range: 3 months-15 years). Pain or painful paraesthesias were the main complaints (13/16). Postoperatively, 9 patients were painfree while 4 improved. Similarly, neurological deficits were favourably influenced by the operation: Out of 5 patients with motor deficits 4 had complete, 1 patient had partial recovery. One out of 4 patients with sensory deficits had complete recovery, 2 remained unchanged, while 1 worsened. Two patients developed new motor and 6 patients new sensory deficits, which (in the course of time) did not disappear completely. New deficits developed predominantly in patients with large tumours or longstanding symptoms. Tumour recurrences were not seen during the follow-up period of 23 months. Our findings revealed that in the majority of cases peripheral nerve neurinomas can be excised with good results. Patients should be treated by a neurosurgeon with special expertise in peripheral nerve surgery. The patient should be thoroughly informed pre-operatively about any eventual new neurological deficits following surgery. PMID- 9401656 TI - Giant intrasacral schwannomas: report of six cases. AB - Only 4 of the 30 previously reported cases of giant sacral schwannomas have been studied with Magnetic Resonance Imaging (MRI). We are reporting 6 more cases, 5 of which had MRI studies. There were 5 women and 1 man (average age 45 years) with long lasting symptoms consisting of lumbosacral and radicular pain accompanied by urinary disturbances and dysaesthetic sensations in the lower limbs. CT clearly defined sacral bone involvement but poorly demonstrated intraspinal tumour extension which was more evident in MRI studies. MRI also clearly showed the intrapelvic extension of the tumour, its relationship with the neighbouring structures and the dumbbell growth pattern due to tumour extension through sacral foramina which are important data for making a pro-operative diagnosis and surgical planning. Surgical treatment consisted of piecemeal tumour resection through a posterior approach in four cases. Two patients underwent operation through an abdominal transperitoneal approach followed by a sacral laminectomy. Total intracapsular resection was apparently achieved in 5 cases. Through an average follow-up period of 9.2 years and despite a rather conservative approach, the recurrence rate has been very low in our series and only one patient had to be re-operated on for tumour recurrence. PMID- 9401657 TI - Primary malignant rhabdoid tumours of the central nervous system: an immunohistochemical and ultrastructural study. AB - Three cases of primary rhabdoid tumour of the CNS (RT-CNS) are presented. In case 1 a hemispheric tumour developed in a 10.5 months old girl, who survived for 6 months after incomplete resection, radio- and polychemotherapy. Case 2 was a 4 years and 8 months old boy with a large IIIrd ventricle tumour, who died of leptomeningeal tumour dissemination 7 months after diagnosis despite radiotherapy. In case 3 a pineal mass occurring in a 14 month old female was radioresistant and totally exstirpated. The child died due to tumour recurrence two months later. Autopsy examination revealed widespread leptomeningeal dissemination. All three cases fulfilled light and electron microscopic criteria of RT-CNS including abundant eosinophilic cytoplasm, vesicular nuclei with large nucleoli and conspicuous anti-vimentin positive filaments. Extensive immunohistochemical studies showed expression of epithelial (EMA, KL1), macrophage (alpha-1 antichymotrypsin), neuro-ectodermal (GFAP, NSE, beta-tubulin III) and myogenic markers (desmin, actin). Different stress proteins (alpha-B crystallin, HSP70) were also expressed. Tumour cells showed a proliferation (MIB1) index of 28.4% (case 1) and 33.4% (case 2). From our study it can be concluded that RT-CNS reveals significant immuno-morphological heterogeneity thus supporting the view that it is not a specific pathological entity but merely a phenotypic appearance of different neoplasms, some of which are linked to primitive neuro-ectodermal tumours. PMID- 9401658 TI - Cranioplasty with autogenous autoclaved calvarial bone flap in the cases of tumoural invasion. AB - When a bone flap is raised in the course of a craniotomy, the ideal is to replace it at the end of the procedure. When it is invaded by tumoural cells, it cannot be replaced due to the risk of tumoural recurrence. In these cases we have autoclaved the bone flap to be able to replace it with no fear of tumoural recurrence. Between October 1989 and October 1995 sixty-two patients required autoclaving of the bone flap in the course of a craniotomy due to tumoural invasion (thirty-five meningiomas, sixteen bone tumours, five metastases, and eight scalp tumours). The infiltrated bone flaps were removed, cleaned, autoclaved for 20 minutes at 134 degrees C and 1 kg/cm2 and re-implanted. Patients were followed-up for 10 to 58 months (average 41 months). At every follow-up visit skull x-ray studies, clinical examination, and photographs were done. When needed a CT scan was performed to assess the thickness of the bone flap. On follow-up roentgenograms partial resorption was observed in twelve cases (19.3%). CT scan studies showed loss of thickness in another thirty-five cases (56.4%). Meanwhile the external aspect remained unchanged. In six cases (3.2%) biopsies of the bone flaps were taken at a second surgical procedure. They showed newly formed bone partly re-populated by osteocytes but retaining areas of sequestered bone. We conclude that autoclaved bone, if replaced with direct contact with living bone, it is gradually repopulated with osteocytes. Cranial vault autoclaved autologous bone flap is a good alternative when the original bone flap is invaded but not destroyed by tumoural cells. PMID- 9401659 TI - Brain spatula with transparent tip: technical note. AB - A newly developed brain spatula with transparent tip for brain retraction is introduced. The high-molecular polymer plastic material is used only for the tip of the new spatula whilst rest of the spatula is made up of the ordinary malleable metal. The transparent nature of the spatula tip helps us in observing the retracted brain vessels and cranial nerves in continuity with the main operating area. The extent of distortion as a result of the retraction can be directly observed assisting in prevention of an inadvertent injury. The new brain spatula with transparent tip is helpful for microneurosurgery under high magnification. Various shapes and sizes of the spatula can be used. PMID- 9401660 TI - Usefulness of non-woven sheet (clip pad) on scalp incisions: technical note. AB - As an alternative to the gauze often used on scalp incisions, we prepared a pad made of non-woven sheets, coated with adhesive on one side. Unlike conventional gauze, the pad did not slip away from the incision line or become entangled with the scalp clips, thus demonstrating its high usefulness for this purpose. PMID- 9401661 TI - Local variations in the cerebral microcirculatory response to hypercapnia and haemorrhage. AB - This study evaluates local variations of the cerebral vasomotor responses to hypercapnia and haemorrhagic hypotension in a pig model. Four laser Doppler flow probes were used in each pig. There was considerable variation in laser Doppler signals between the four probes in baseline recordings. The increases in flow after CO2 administration in 7 pigs had a mean coefficient of variation of 0.43 +/ 0.31, and the flow changes after blood loss in another 7 pigs had a mean coefficient of variation of 0.45 +/- 0.34. The range of flow changes within each animal was large; the probe with the highest CO2 response showed on the average a 273% +/- 157% larger CO2 response than the probe with the lowest CO2 response. Correspondingly, the probe with the best preserved blood flow after blood loss had on the average a flow value of 93% +/- 12% of the baseline value, while the probe that changed most with haemorrhage had a flow value of 44% +/- 24% of the baseline value. Single laser Doppler recordings have been used for the monitoring of cerebral blood flow in neurosurgical critical care, but our results suggest that a single laser Doppler flow probe is not an adequate method to monitor vasoreactivity in neurosurgical patients because flow signals from one probe may be unrepresentative for other sites in the brain. PMID- 9401662 TI - Initial and postoperative hyponatremia associated with pituitary adenoma: a case report. AB - This 67 year-old man experienced 3 episodes of symptomatic hyponatraemia. Radiological examination revealed a sellar lesion and the tumour was removed via the transsphenoidal route. Thereafter, he simultaneously developed intractable diabetes insipidus and serious hyponatraemia with persistent natriuresis. His level of atrial natriuretic peptide was not significantly elevated, however, his plasma aldosterone concentration was low. The oral administration of salt gradually improved his hyponatraemia as well as the coincident symptoms. By the administration of a mineralocorticoid, fludrocortisone acetate, we succeeded in maintaining his serum sodium level without salt replacement. We discuss the mechanism(s) and treatment of hyponatraemia associated with pituitary tumour. PMID- 9401663 TI - Pituitary apoplexy with spontaneous cure of acromegaly and its possible relation to Gd-DTPA-administration. AB - A case of pituitary apoplexy occurring after Gd-DTPA-administration for contrast enhanced MRI in a patient with an hGH-producing macro-adenoma is presented. Within days the initially increased hGH level fell to the normal range, the oral glucose tolerance test (OGTT) showed a normal suppression of hGH and complete anterior pituitary insufficiency developed. At this time repeated MRI suggested a haemorrhagic infarction of the macro-adenoma. Fourteen months later re examination confirmed spontaneous cure of the acromegaly, improvement of adenopituitary function and shrinkage of the sellar content. The causal linkage between the pituitary adenoma apoplexy and Gd-DTPA-administration is unclear. It might be due to contrast induced blood pressure and endothelial permeability changes, possibly promoted by pre-existing diabetes mellitus associated vasculopathy. PMID- 9401664 TI - Fulminant cerebral infarctions caused by nonbacterial thrombotic endocarditis due to gallbladder cancer. PMID- 9401665 TI - Genotype- and gender-dependent hepatic alcohol dehydrogenase (ADH) activity in developing mice. AB - Ethanol is metabolized in human and rodent liver primarily by the enzyme alcohol dehydrogenase (ADH). Hepatic ADH activity in adult males and females of seven inbred strains of mice was determined to examine genotype- and sex-dependent variability among the strains and the level of sexual dimorphism in each of the strains. ADH activity varied considerably among the strains, which could be categorized into high-activity strains C57BL/6, C57Brcd, and Swiss, and relatively low-activity strains C3H, CBA, and DBA. Adult Swiss, AKR, C57BL/6, and DBA females exhibited significantly higher levels of hepatic ADH than their male counterparts, whereas no gender differences were seen in C3H, CBA, and C57Brcd. Young mice of high and/or low ADH activity strains viz. C57BL/6 and C3H did not exhibit gender differences in ADH activity at weanling but the enzyme levels increased by sixth week in females and remained higher thereafter. The progeny of a high-activity strain with sexual dimorphism (C57BL/6) and a low-activity strain lacking gender difference (C3H) exhibited intermediate levels of ADH and age dependent sexual dimorphism. PMID- 9401666 TI - Ethanol and glutamate effects on intracellular magnesium. AB - Acutely dissociated rat brain cells were loaded with the magnesium-sensitive fluorescent dye furaptra. Stimulation with varying concentrations of GLU + 1 microM GLY produced a concentration-dependent increase in free intracellular magnesium ([Mg2+]i) that was not dependent on the presence of extracellular Mg2+. Ethanol (50 mM) did not significantly inhibit GLU/GLY-stimulated increases in [Mg2+]i. However, ethanol alone significantly increased baseline [Mg2+]i. PMID- 9401667 TI - Dexmedetomidine alleviates ethanol withdrawal symptoms in the rat. AB - The effect of dexmedetomidine, a selective alpha 2-adrenoceptor agonist, on ethanol withdrawal symptoms was studied in chronically ethanol-fed rats. After a 4-day ethanol intoxication period the rats were given s.c. injections of dexmedetomidine (3, 10, or 30 micrograms/kg) or saline (control group) at 10, 16, 22, and 39 h after the last dose of ethanol. The severity of ethanol withdrawal symptoms (rigidity, tremor, irritability, hypoactivity) was rated up to 58 h, blind to the treatments. The results showed that dexmedetomidine at doses 10 and 30 micrograms/kg significantly diminished the severity of the ethanol withdrawal reaction as measured by the sum score of the three most specific withdrawal signs (rigidity, tremor, and irritability). Dexmedetomidine at 10 micrograms/kg was the most effective dose, especially in the latter half of the withdrawal period (23 58 h after last dose of ethanol). The results suggest that dexmedetomidine in the treatment of ethanol withdrawal symptoms should be further studied. PMID- 9401668 TI - Total contractile protein contents and gene expression in skeletal muscle in response to chronic ethanol consumption in the rat. AB - An investigation was carried out to determine changes in the contents of skeletal muscle myofibrillary proteins (i.e., the contractile fraction composed principally of actin and myosin) and gene expression in skeletal muscle in response to ethanol feeding. Male Wistar rats were fed a nutritionally complete liquid diet, which contained 35% of total calories as ethanol. Controls were pair fed isocaloric amounts of the same diet, in which ethanol was replaced by isocaloric glucose. Total mixed and contractile protein contents of the gastrocnemius in ethanol-fed rats were rapidly reduced by ethanol feeding: a response was discernible as early as 1 week after the commencement of the ethanol feeding regimen (approx. -10%, p < 0.025 and p = 0.05 for mixed and myofibrillary proteins, respectively). At 2, 4, and 6 weeks, mixed and myofibrillary protein contents were further reduced in alcohol-fed rats, by between 12% and 22%, compared to pair-fed controls. Similar changes occurred in the soluble (i.e., sarcoplasmic) protein fractions of skeletal muscle. At 2 weeks the composition of total messenger RNA and individual messenger RNA species was measured. Total messenger RNA content per muscle was reduced by 35% (p < 0.05). Messenger RNA levels for alpha-actin, beta-myosin heavy chain, and carbonic anhydrase III were not significantly altered. In conclusion, skeletal muscle protein contents are rapidly reduced by ethanol feeding, compared to pair-fed controls, though mRNA species encoding specific isoforms of myosin and actin are not affected. It is possible that chronic ethanol feeding may significantly alter the stability of mRNAs encoding other contractile proteins, or alternatively, defects in translation may predominate. PMID- 9401670 TI - Chronic low doses of ethanol affect brain protein kinase C and ultrasonic calls in rats. AB - Few studies have investigated neurobehavioral and neurochemical consequences of chronic consumption of low doses of ethanol. The present study shows that in rats exposure to 3% ethanol (v/v in drinking water) for 2 months decreased both calcium-dependent and -independent protein kinase C (PKC) activities in the cortex and in the hippocampus. This treatment also reduced ultrasonic calls (UCs), an index of emotional and motivational states of the animal. In addition, at cortical level of ethanol-treated rats, we observed a correlation between calcium-dependent activities and UCs. These results suggest that nonaddicting doses of ethanol affect brain PKC activities and that this enzyme may be involved in the ethanol modulation of emotional and motivational behaviors. PMID- 9401669 TI - The role of ethanol availability on stress-induced increases in ethanol consumption. AB - Exposure to stressors can increase ethanol consumption and ethanol can attenuate the behavioral and biochemical effects of stressors. This study determined whether the availability of ethanol during the period of exposure to repeated restraint alters the poststress increase in ethanol intake. Seven days of restraint increased ethanol intake on the first poststress test only in animals deprived of ethanol during the restraint period. These results indicate that the availability of ethanol during exposure to restraint can attenuate the impact of restraint on ethanol intake. Ethanol intake was also positively related to novelty-induced locomotion and restraint eliminated this relationship. However, amphetamine-induced locomotion was not altered by either restraint or EtOH intake. These results indicate that voluntary ethanol intake can attenuate the impact of restraint stress and that restraint stress can alter the influence of ethanol on novelty-induced locomotion. It is suggested that this symmetrical relationship between ethanol intake and restraint stress may be involved in an interactive manner that determines stress-induced ethanol consumption. PMID- 9401671 TI - Biochemical and histopathological changes in arsenic-intoxicated rats coexposed to ethanol. AB - The effects of arsenic and ethanol interaction on blood, liver and serum biochemical indices, and arsenic concentration in soft tissues of rats were investigated to determine the influence of these substances in inducing susceptibility to arsenic poisoning. Arsenic, intraperitoneally (100 ppm, once, daily), ethanol in drinking water (10%), or the combination were administered for a period of 6 weeks. Both the chemicals had some additive effects in marginally elevating blood zinc protoporphyrin. Glutathione (GSH) concentrations of blood and liver were reduced by both arsenic and ethanol; however, there was a more pronounced depletion of hepatic GSH concentration in animals coexposed to arsenic and ethanol. Combined arsenic plus ethanol exposure led to significantly more elevated activities of serum transaminases than in animals administered arsenic or ethanol alone. Histopathological alterations in kidneys and liver occurred following arsenic exposure. Arsenic plus ethanol produced more pronounced liver lesions, whereas kidney changes were the same as with arsenic alone. The concentrations of arsenic in kidney and liver were higher in rats exposed to arsenic plus ethanol. The results suggest that animals exposed to arsenic plus ethanol are more vulnerable to arsenic toxicity. PMID- 9401672 TI - Prenatal exposure to ethanol in rats: effects on liver energy level and antioxidant status in mothers, fetuses, and newborns. AB - The fetal alcohol syndrome is a clinical condition that affects newborns from alcoholic mothers. It is not clear, however, whether ethanol consumption during gestation can affect liver functions of fetuses and newborns. In this study, we aimed to assess the effects of ethanol administration on body weight, liver energy level, and antioxidant status of mothers, fetuses, and newborns. Pregnant rats were exposed to ethanol during the third week of gestation. Body weight, survival, and liver concentration of gluthatione (GSH) and adenosintriphosphate (ATP) were measured. No differences were observed in body weight or in liver ATP and GSH between mothers exposed to ethanol and control animals. Conversely, fetuses from rats exposed to ethanol showed a marked decrease in GSH, ATP, and body weight when compared to those from control rats. Newborns exposed prenatally to ethanol were no different from those born to control mothers. This study suggests that an amount of ethanol that is not sufficient to determine a significant effect on mothers can, nevertheless, cause a marked decrease in growth and in liver antioxidant and energy status in fetuses. These parameters, however, return to control value one week after ethanol discontinuation. PMID- 9401673 TI - Developmental changes in the inhibitory actions of ethanol on glutamate-induced translocation of protein kinase C in cerebellar granule neurons. AB - The effects of increasing concentrations of ethanol (25-200 mM) on the enhancement of [3H]phorbol-12,13-dibutyrate ([3H]PDBu) binding produced by different glutamate receptor agonists, indicative of a translocation of the intracellular enzyme protein kinase C (PKC), were studied in rat cerebellar granule cells at 2, 4, 8, and 12 days in vitro (DIV). Glutamate-produced stimulation of [3H]PDBu binding was inhibited by 50 mM ethanol at 2 DIV, whereas higher ethanol concentrations (> 100 mM) were needed to reduce the increase of [3H]PDBu binding in cells grown for 4, 8, and 12 DIV. Ethanol significantly inhibited NMDA-stimulated [3H]PDBu binding in a concentration-dependent fashion in cells maintained in culture for 4 and 8 days, respectively, with a slightly less pronounced inhibition by ethanol (50 mM) seen in cells kept for 2 and 12 DIV. Application of higher ethanol concentrations (> 100 mM), inhibited the NMDA induced stimulation in all cell preparations. Following kainic acid-induced enhancement of [3H]PDBu binding, ethanol (100 mM) reduced the binding only in cells maintained for 2 DIV. Even higher ethanol concentrations (200 mM) inhibited the effects of kainic acid only in cells maintained for 2 and 4 DIV, respectively. Our data suggest that various subclasses of glutamate receptors display a developmentally determined differential sensitivity to ethanol at least in cerebellar granule cells in vitro. PMID- 9401674 TI - Effect of ethanol drinking and naltrexone on subsequent drinking in rats. AB - The effects of several days of oral ethanol drinking paired with naltrexone (NTX) on subsequent ethanol drinking were investigated in rats. We hypothesized that repeated pairings of NTX combined with forced oral ethanol intake would extinguish ethanol drinking so that when NTX injections were terminated, voluntary oral ethanol drinking would be suppressed. Thirty-two male. Long-Evans rats were provided with alternate days of either 8% ethanol solution or water as the sole source of fluid. Intraperitoneal injections of 0, 2.5, or 5.0 mg of NTX hydrochloride were administered on the ethanol days. Following the termination of injections, rats were returned to unrestricted access to water and ethanol and 24 h measurements of fluid intake were recorded. NTX decreased ethanol intake 4 h, but not 24 h, after NTX injections. Despite the consumption of significant amounts of ethanol during NTX treatment, there was no change in voluntary oral ethanol intake patterns after NTX injections were terminated (reinstatement of voluntary ethanol drinking). Thus, NTX's reduction in ethanol intake was limited in duration and did not result in long-term extinction of ethanol drinking behavior. PMID- 9401675 TI - Sulpiride-induced increases in serum prolactin levels in female rats exposed prenatally to alcohol. AB - We examined the impact of prenatal alcohol exposure on serum prolactin levels and on the ability of the D2 dopamine antagonist sulpiride to stimulate prolactin release in Long-Evans rats. Pregnant rats were intubated with alcohol (0, 3, or 5 g/kg/day) from gestational day 8 (GD8) to GD20. Adult female offspring were screened for estrous cycle stage. At diestrus, the rats were challenged with a single dose of sulpiride (0, 10, or 40 micrograms/kg) and trunk blood was collected 20 min later. After prenatal exposure to either dose of alcohol, mean basal serum levels of prolactin were about 65% less than the 0 g/kg group, and the 35-40% mean differences from an untreated control group were not significant. Sulpiride produced dramatic dose-dependent increases in serum prolactin levels in all prenatal treatment groups. Across all doses of sulpiride, the group given the higher dose of prenatal alcohol (5 g/kg/day) had significantly lower serum prolactin levels than all other groups. There was no significant interaction between prenatal treatment and sulpiride dose. Neither prenatal alcohol exposure nor sulpiride injections had significant effects on serum corticosterone levels in this study. Although the current results are unclear regarding a baseline decrease in prolactin levels after prenatal alcohol exposure, the overall results suggest that prenatal alcohol exposure decreases prolactin levels but there is no evidence that it does so by altering dopaminergic tone in hypothalamus of female rats. PMID- 9401676 TI - Effects of ethanol on striatal dopamine overflow and clearance: an in vivo electrochemical study. AB - Previous studies have shown that the neurotransmitter dopamine (DA) is implicated in the reinforcing effects of ethanol and other abused drugs. Ethanol also alters DA overflow and uptake in vivo. Further studies of the role of DA in the behavioral and neurochemical effects of ethanol may help explain the pharmacological mechanisms by which these effects are produced. In these studies we used in vivo electrochemical recordings to investigate the effects of ethanol (EtOH) on the dynamics of evoked DA overflow and DA uptake in rat dorsal striatum. Local applications of EtOH from a multibarrel micropipette did not produce detectable changes in extracellular levels of endogenous DA in the dorsal striatum. EtOH application did attenuate potassium (K+)-evoked overflow of DA in a time-dependent fashion. In contrast, tyramine-induced DA overflow, a calcium independent process thought to be carrier mediated, was not altered by local EtOH application in the dorsal striatum. Striatal uptake of locally applied exogenous DA was decreased by nomifensine, an effect that was attenuated by locally applied EtOH. Taken together, these data suggest that one of the effects of EtOH on DA containing nerve endings in the rat striatum involves functional changes in the high-affinity DA transporter associated with these nerve endings. PMID- 9401678 TI - Sardinian alcohol-preferring rats prefer chocolate and sucrose over ethanol. AB - The present study was aimed at determining whether the concurrent availability of highly palatable fluids (i.e., a chocolate-flavored drink and a sucrose solution) would alter voluntary ethanol drinking in selectively bred, alcohol-preferring sP and -nonpreferring sNP rats. Ethanol intake occurred under the three-bottle, free choice regimen between 10% (v/v) ethanol solution, tap water, and the palatable fluids for 24 h per day. When rats were given ethanol and water, but no alternative fluids, mean ethanol intake in sP rats ranged between 6 and 7 g/kg per day and mean preference ratio was steadily higher than 80%, whereas mean ethanol intake and preference ratio in sNP rats were constantly lower than 0.3 g/kg and 5%, respectively. In the presence of either the chocolate-flavored drink or sucrose solution, both prepared as isocaloric to the ethanol solution, absolute ethanol intake in sP rats declined by 60-70%; similarly, the preference ratio was reduced by 80-90%. Ethanol intake in sNP rats was unaffected by the simultaneous presentation of either palatable fluids. The results of the present study closely replicate those previously reported in genetically selected, ethanol-preferring HAD rats; however, they differ from those of ethanol preferring P rats, which were reported to maintain high levels of ethanol intake and preference in the presence of highly palatable fluids. These results are discussed in terms of a) an alternative reinforcement partially substituting for the reinforcing properties of ethanol in sP rats, resulting in a less urgent need of ethanol, and b) genetic animal models of alcoholism diverging in some neurochemical and behavioral traits (e.g., response to the presentation of palatable fluids), which might parallel the different types of alcoholism observed in humans. PMID- 9401677 TI - Regional CNS densities of serotonin and dopamine receptors in high alcohol drinking (HAD) and low alcohol-drinking (LAD) rats. AB - The densities of subtypes of serotonin (5-HT) and dopamine (DA) receptors were determined in the CNS of male alcohol-naive HAD and LAD lines of rats. Autoradiographic studies were undertaken to measure the densities of (a) 5-HT1A sites labelled with 2 nM [3H]8-OH DPAT, (b) 5-HT2A sites labelled with 2 nM [3H] ketanserin, (c) D1 sites labelled with 1 nM [3H]SCH23390, and (d) D2 sites labelled with 20 nM [3H]sulpiride. Membrane binding, using tissue combined from the olfactory bulb, olfactory tubercle, and nucleus accumbens, was carried out to determine Kd and Bmax values for the binding of 0.25-8.0 nM [3H]7-OH DPAT to D3 sites. Among the 14 regions measured for densities of 5-HT1A sites, no interline differences were found in the cerebral cortical regions or in the septal nuclei; however, within the hippocampus, 15-20% lower binding of [3H]8-OH DPAT was observed in the posterior dorsal CA3 and dentate gyrus of the HAD line. There were no interline differences in any of the 10 regions examined for [3H]ketanserin binding to 5-HT2A sites, or in the densities of D1 and D2 sites in the mesolimbic and nigrostriatal DA systems, except for a 35% higher density of D2 sites in the substantia nigra pars compacta of the HAD line. There were no interline differences in the Kd or Bmax values for [3H]7-OH DPAT binding to D3 sites. Overall, these results indicate that no marked interline differences are evident in the densities of 5-HT1A, 5-HT2A, D1, D2, and D3 receptors within the mesolimbic system that could be associated with the disparate alcohol drinking behaviors of the HAD and LAD rats. PMID- 9401679 TI - Adenosine A1 receptor antisense infused in striatum of rats: actions on alcohol induced locomotor impairment, blood alcohol, and body temperature. AB - Previous pharmacological studies show that adenosine receptors in the corpus striatum may be involved in locomotor coordination. The purpose of this investigation was to determine whether the adenosine A1 receptor subtype would alter the locomotor response due to incapacitating doses of alcohol. In these experiments, an antisense oligodeoxynucleotide (ODN) targeted to the adenosine A1 receptor was used to elucidate its possible role in locomotor function. After bilateral cannulation of the caudate nuclei of two strains of adult male rats, the animals were trained to remain on a rotorod for an entire 3-min interval. Then, a standard dose of 2.0 micrograms per 2.0 microliters of the A1 adenosine antisense (A1AS), dissolved in a pyrogen-free artificial cerebrospinal fluid (aCSF), was microinjected four times bilaterally into the caudate nuclei of the rats at successive 12-h intervals over 2 days. Three sets of controls were utilized: intragastric gavage with tap water alone: intragastric gavage of 3.5 4.0 g/kg 20% alcohol alone; and the aCSF vehicle alone microinjected identically in the caudate nuclei. The results showed that the intragastric administration of 20% alcohol in a dose of 3.5-4.0 g/kg caused a complete incapacitation of locomotor performance. Moreover, the A1AS injected in the striatum failed to alter significantly the action of alcohol in its impairment of the rats' ability to negotiate the rotorod. Concurrent measures of blood alcohol and body temperature taken to validate the efficacy of alcohol administration correlated precisely with the blockade of locomotor behavior of the animals. These findings thus suggest that because of the specificity of the A1AS probe, the A1 receptor in the striatum is not involved in the alcohol-induced incapacitation of locomotor activity. PMID- 9401680 TI - Accuracy of localizing radiopaque markers by abdominal radiography and correlation between their gastric emptying rate and that of a canned food in dogs. AB - OBJECTIVES: To determine accuracy of abdominal radiography in locating radiopaque markers in the gastrointestinal tract and to assess correlation between gastric emptying rate of radiopaque markers and that of canned food. ANIMALS: 17 healthy dogs. PROCEDURE: Dogs were fed thirty 1.5-mm markers and ten 5-mm markers mixed in sufficient food to meet 25% of their daily caloric intake. They were then euthanatized by administration of an overdose of barbiturate at 1, 2, 5, 8, or 12 hours after eating and the abdomen was radiographed. The stomach, small intestine, and large intestine were then separated and radiographed in isolation. The wet and dry weights of the stomach contents were determined. The apparent and actual locations of the markers and the gastric emptying rates of markers, wet matter, and dry matter were compared, using rank correlation. RESULTS: All comparisons indicated significant (P < 0.025), high correlation coefficients (> 0.92). The mean difference between the apparent and actual locations of the markers was < 3% for all comparisons. The mean difference between the percentage of small markers and large markers retained in the stomach and that of dry matter was 7.8 (SD, 6.2; range, 0 to 18%) and 11.9 (SD, 12.5; range, 0 to 44%), respectively. CONCLUSIONS: The gastric emptying and orocolic transit rates of the markers were accurately predicted by abdominal radiography. The gastric emptying rate of the diet and the small markers and, to a lesser extent, the large markers was closely correlated. CLINICAL RELEVANCE: When fed with a special canned food diet, radiopaque markers can be used to assess the gastric emptying rate of food with sufficient accuracy for clinical purposes. PMID- 9401681 TI - Detection of antibodies to Aspergillus fumigatus in serum of horses with mycosis of the auditory tube diverticulum (guttural pouch). AB - OBJECTIVE: To detect antibodies against Aspergillus fumigatus antigens in serum samples from horses and to evaluate the relevance of this method as an alternative approach to the diagnosis of mycosis of the auditory tube diverticulum (guttural pouch mycosis [GPM]). ANIMALS: Twelve clinically normal horses (controls) and 12 horses with GPM diagnosed by endoscopic observation of characteristic mycotic plaques. PROCEDURE: Antibodies to A fumigatus antigens were detected in serum by use of an ELISA and immunoblot analysis with extracellular antigens. RESULTS: Antibodies against A fumigatus antigens were found in healthy and diseased horses. Titer of total Aspergillus antibodies was not diagnostic for GPM. In contrast, immunoblot analysis results indicated that 2 antigens of 22 and 26 kd were constantly recognized by sera from diseased horses. CONCLUSIONS: Reactivity to 22- and 26-kd A fumigatus antigens, as measured by immunoblot analysis, seemed to be diagnostic for GPM in horses. PMID- 9401682 TI - Comparison of radiography, magnetic resonance imaging, and surgical findings in dogs with elbow dysplasia. AB - OBJECTIVE: To describe the magnetic resonance imaging (MRI) appearance of medial coronoid process and humeral condyle lesions in dysplastic cubital joints and to compare survey radiography and MRI for evaluation of fragmented medial coronoid process (FMCP) and lesions of the medial aspect of the humeral condyle (MAHC). ANIMALS: 18 dogs with elbow dysplasia. PROCEDURE: Radiography of 22 cubital joints was performed. The 22 joints then underwent MRI. The scans were evaluated with regard to the shape and signal of the coronoid process; articular cartilage change, subchondral bone disruption of the MAHC. Surgical findings were used as the standard to calculate accuracy, sensitivity, specificity, and positive- and negative-predictive values for specific diagnosis of FMCP (free fragment) and lesions of the MAHC. RESULTS: At surgery, 31.8% of the joints had FMCP (free), 36.4% had nondisplaced unmineralized coronoid process, and 27.2% had nondisplaced mineralized coronoid process. Eleven joints had lesions of the MAHC, and wear lesions were observed in 41% of the joints. On radiography, FMCP (free) was visualized in 9% of the joints and lesions of the MAHC were observed in 23%. MRI had the highest accuracy (95.5%), sensitivity (100%), and negative-predictive value (100%) for detection of FMCP (free), and had accuracy (91%), sensitivity (87.5%), specificity (92.5%), and positive (87.5%)- and negative (92.5%) predictive values for detection of nondisplaced unmineralized coronoid process. CONCLUSIONS AND CLINICAL RELEVANCE: Compared with radiography, MRI was useful for detection of nondisplaced unmineralized coronoid process; images consistently correlated with surgical findings. The technique is accurate and especially useful when radiographic findings are inconclusive. PMID- 9401683 TI - Immunohistochemical diagnosis of pestivirus infection associated with bovine and ovine abortion and perinatal death. AB - OBJECTIVE: To establish a reliable, rapid, economical method for detection of pestivirus infection in bovine and ovine fetuses and to examine participation of these viruses in abortions and neonatal mortality. ANIMALS: 213 bovine and 31 ovine fetuses, as well as 36 newborn calves and 25 lambs, which had died within 3 days after birth, were tested for bovine viral diarrhea virus (BVDV) and border disease virus by use of different methods. PROCEDURE: Detection of BVDV in fetuses was performed by immunohistochemical methods, using a panel of monoclonal antibodies against pestivirus antigens on cryostat and paraffin sections and by virus isolation in cell culture; in some instances, an antigencapture ELISA was performed. Results of the various methods were compared. RESULTS: Sensitivity of BVDV detection by immunohistochemical methods and virus isolation in cell culture was equal; however, it decreased in association with autolysis. In autolytic fetuses, use of formalin-fixed, paraffin-embedded brain sections was the most favorable method. Antigen detection by ELISA was less sensitive. CONCLUSIONS: Immunohistochemical analysis of cryostat sections of brain, skin, thyroid gland, abomasum, and placenta is a rapid, sensitive method for detecting pestiviruses in fetuses. In the presence of advanced autolysis, this method used on formalin fixed, paraffin-embedded brain sections is recommended over the other described methods. PMID- 9401684 TI - Evaluation of platelet activation and platelet-neutrophil aggregates in ponies with alimentary laminitis. AB - OBJECTIVES: To determine whether platelets are hyperaggregable or form platelet neutrophil aggregates during the prodromal stages of acute laminitis of ponies. ANIMALS: Healthy adult ponies: 8 experimental and 6 control. PROCEDURES: Acute laminitis was induced by oral administration of corn starch and wood flour to 8 ponies, and indices of platelet activation were evaluated. Blood samples were collected before and at 4, 8, 12, 24, 28, and 32 hours after carbohydrate administration, and PCV, total plasma protein concentration, platelet count, activated clotting time, whole blood recalcification time, spontaneous platelet aggregation, ex vivo platelet aggregation responses, and platelet-neutrophil aggregates were determined. When lameness was first detected, ponies were euthanatized and arteriography and histologic examination of hooves were performed. RESULTS: Carbohydrate overload was associated with hyperaggregability of platelets throughout the prodromal stages of laminitis and increased numbers of platelet-neutrophil aggregates. Reduction of blood supply to affected hooves was variable, and blood clots were found in 6 of 11 laminitis-affected hooves. CONCLUSIONS: Platelets were hyperaggregable throughout the prodromal stages of carbohydrate-induced laminitis and formed platelet-neutrophil aggregates. Platelet-neutrophil aggregates may initiate or contribute to development of acute laminitis. CLINICAL RELEVANCE: Anti-platelet therapy may be useful for treatment of acute alimentary laminitis in horses. PMID- 9401685 TI - Effect of congenitally acquired Neospora caninum infection on risk of abortion and subsequent abortions in dairy cattle. AB - OBJECTIVES: To estimate the extent to which abortion risk in dairy cattle during subsequent pregnancies was associated with congenitally-acquired Neospora caninum infection and previous abortions. ANIMALS: 468 Holstein cattle. PROCEDURE: Newborn heifer calves were tested for evidence of congenital infection attributable to N caninum and examined repeatedly until the completion of their second lactation for serologic status and evidence of abortion. RESULTS: Compared with noninfected cows, congenitally infected cows had a 7.4-fold higher risk of abortion during their initial pregnancy and a 1.7-fold higher risk of aborting the first pregnancy during their first lactation. During the first pregnancy of their second lactation, congenitally infected cows that had aborted previously had a 5.6-fold higher risk of abortion, compared with cows that had not previously aborted and that were seronegative. The fetal risk period for N caninum-associated death began sooner and extended later during the initial pregnancy compared with subsequent pregnancies. CONCLUSION: Congenitally acquired N caninum infection can cause a substantial number of abortions during the initial pregnancy of heifers, with abortion risk attributable to N caninum decreasing in subsequent pregnancies, possibly because of selective culling. Subsequent abortions can be expected in congenitally infected cows that have aborted previously. PMID- 9401686 TI - Pulmonary and neutrophil responses to priming effects of platelet-activating factor in pigs. AB - OBJECTIVE: To investigate whether platelet-activating factor (PAF) primes the porcine pulmonary response to lipopolysaccharide (LPS), and what effect PAF priming has on porcine neutrophil superoxide (SO) release. ANIMALS: 8- to 10-week old pigs. PROCEDURES: After pigs were anesthetized with sodium pentobarbital and instrumented for standard cardiopulmonary hemodynamic measurements, they were randomly assigned to receive PAF (0.1 ng/kg of body weight/min, 0 to 2 hours) plus saline solution (2 to 6 hours), saline solution (0 to 2 hours) plus LPS (0.25 microgram/kg/h, 2 to 6 hours), or PAF plus LPS. Cardiopulmonary variables were measured throughout the study. Neutrophils were isolated from saline- or PAF treated pigs at 0 (baseline) and 2 hours, and the effect of in vivo PAF exposure on ex vivo phorbol myristate acetate (PMA)-induced SO release was measured. Additionally, neutrophils isolated from immune-naive pigs were primed in vitro for 10 minutes with 10 microM PAF, and PMA-induced SO release was measured. RESULTS: PAF infusion significantly enhanced the increase in mean pulmonary arterial pressure, pulmonary vascular resistance, and hypoxemia associated with LPS administration. The infusion increased ex vivo neutrophil SO release, and in vitro PAF exposure primed neutrophils for enhanced SO release that was inhibited by pretreatment of cells with indomethacin. CONCLUSIONS: PAF primes the porcine pulmonary system for the response to LPS. It primes porcine neutrophils in vivo and in vitro for PMA-induced SO release, and in vitro priming is mediated by cyclooxygenase products of arachidonic acid metabolism. CLINICAL RELEVANCE: PAF may modulate the porcine inflammatory response by acting as a priming agent, making pigs more responsive to the negative effects of bacterial LPS. PMID- 9401687 TI - Functional and phenotypic characterization of monoclonal antibodies to bovine L selectin. AB - OBJECTIVE: To characterize 2 bovine neutrophil monoclonal antibodies (MAB) as to effects on bovine neutrophil function and their binding antigens on the cell surface of bovine neutrophils. ANIMALS: 16 healthy, lactating Holstein cattle, 1 calf with leukocyte adhesion deficiency, and 1 age-matched control calf, 2 healthy ewes, and 2 healthy human beings as neutrophil sources. PROCEDURE: Neutrophil chemotactic and respiratory burst activities and calcium influx, and binding properties of the 2 MAB were determined. Molecular mass of corresponding cell surface antigens also was determined, as was binding of human L-selectin MAB DREG56 to molecules recognized by MAB 11G10 and 2G8 on the surface of bovine neutrophils. RESULTS: MAB 11G10 and 2G8 inhibited chemotactic activity of bovine neutrophils, up-regulated amplitude of native chemiluminescence, and shortened the time to reach maximal chemiluminescence induced by serum-opsonized zymosan. Crosslinking both MAB with a second antibody induced rapid increase in intracellular free calcium concentration. Binding density of MAB 11G10 and 2G8 to bovine neutrophils treated with trypsin was increased (P < 0.05), compared with that of untreated neutrophils. Neutrophils treated with phosphatidylinositol specific phospholipase C had decreased (P < 0.05) binding density of MAB 11G10 and 2G8. Binding of the various MAB to neutrophils from calves with bovine leukocyte adhesion deficiency was lower (P < 0.05) than binding to neutrophils from healthy calves. Expression of antigens recognized by the aforementioned MAB on the surface of bovine neutrophils was decreased (P < 0.05) within 10 minutes. CONCLUSION: MAB 11G10 and 2G8 recognized L-selectin molecules on bovine neutrophil membrane. L-Selectin (CD62L) is involved in low-affinity adhesion reactions between leukocytes and L-selectin ligand on postcapillary venular endothelial cells. PMID- 9401688 TI - Infection of bone marrow macrophages by equine infectious anemia virus. AB - OBJECTIVE: To characterize infection of bone marrow-derived macrophages (BMDM) with equine infectious anemia virus (EIAV) by determining virus production, effects on viability, and induction of cytokines. SAMPLE POPULATION: BMDM obtained from bone marrow of 6 clinically normal adult horses. PROCEDURE: BMDM were infected with EIAV at a multiplicity of infection of 8. Cell viability, percentage of cells with detectable viral protein, reverse transcriptase activity, and concentrations of infective virus (focus-forming units/ml), interleukin 6, and tumor necrosis factor-alpha were measured in culture supernatant samples obtained at various days after infection. RESULTS: Cell viability was decreased on day 4 and was maximally decreased on day 8. The number of cells with detectable viral protein and supernatant reverse transcriptase activity increased significantly on day 4 and increased until day 6. Virus concentration (focus-forming units per milliliter) peaked on day 4 after infection and was constant thereafter. Infection with EIAV caused significant induction of interleukin 6 production by BMDM. The maximal difference was seen on day 4 after infection. Control and infected BMDM produced only negligible amounts of tumor necrosis factor-alpha. CONCLUSIONS: BMDM are useful, as a cell population, to study the effects of infection with EIAV, including cell death and induction of interleukin 6 but not tumor necrosis factor-alpha production. PMID- 9401689 TI - Quantification of antigen-specific antibody concentrations in tracheal lavage fluid of horses with summer pasture-associated obstructive pulmonary disease. AB - OBJECTIVE: To determine whether horses with summer pasture-associated obstructive pulmonary disease (SPAOPD) have increased concentrations of antigen-specific IgG and IgE in tracheal lavage fluid, compared with values in clinically normal horses. ANIMALS: 8 horses (6 females, 2 geldings; 6 Quarter Horses, 2 Appaloosas), 14 to 23 years old and with previous diagnosis of SPAOPD, served as the principal group; 8 horses (2 females, 6 geldings; 1 Quarter Horse, 7 Thoroughbreds), 6 to 9 years old, with no evidence of respiratory tract disease, served as the control group. PROCEDURE: Data were collected twice during a 1-year period: when all SPAOPD-affected horses were manifesting clinical signs of disease (July), and when all SPAOPD-affected horses appeared clinically normal (February). On each occasion, clinical evaluations were performed and blood and tracheal lavage fluid samples were collected. Transtracheal lavage supernatant was evaluated for mold antigen-specific IgG and IgE concentrations. RESULTS: Median IgE relative antibody unit (RAU) values were significantly higher in control, compared with principal, horses. The SPAOPD-affected horses had increased concentrations of specific IgG for only 1 antigen, during winter sample collection. CONCLUSION: Antigen-specific IgG and IgE RAU values were not increased in SPAOPD-affected horses when these horses were manifesting clinical signs of disease. PMID- 9401690 TI - Comparison of pulsed-field gel electrophoresis and phage typing for discriminating poultry strains of Staphylococcus aureus. AB - OBJECTIVE: To compare pulsed-field gel electrophoresis (PFGE) patterns of Staphylococcus aureus from chickens in England, Belgium, Bulgaria, Argentina, and Japan, to assess the value of PFGE for discriminating strains, and to compare results obtained by PFGE with those obtained by biotyping and phage typing. SAMPLE POPULATION: 78 S aureus isolates from diseased and healthy chickens. PROCEDURE: Chromosomal DNA of S aureus was digested with restriction endonuclease Sma I, and fragments were separated by PFGE in 1% agarose gel. RESULTS: All 78 strains from 5 countries were classified as poultry ecovar according to a previously established biotyping system. Chromosomal DNA was cut by Sma I into 18 to 23 fragments ranging from about 3 to 685 kb. Seventy-eight strains produced 15 types, arbitrarily designated A to O, and 45 subtypes. Some differences were observed in PFGE patterns among countries. However, 10 fragments (333, 190, 110, 63, 55, 42, 34, 19, 10, and 3 kb) were highly conserved and were shared by almost all (> 78%) of the strains examined. The PFGE patterns were compared with those obtained by phage typing. All 29 strains belonging to avian phage-group II produced type A and 19 subtypes. Of the 15 strains belonging to phage-group I, 11 produced 8 types (B to H, O) and 5 subtypes that were different from those of type A. CONCLUSIONS: Genomic DNA fingerprinting by PFGE is an effective technique for discriminating poultry S aureus strains and appears to be a useful method for subtyping strains of avian phage groups or the poultry-specific ecovar. PMID- 9401691 TI - Diagnosis of trichinellosis in swine by enzyme immunoassay, using a synthetic glycan antigen. AB - OBJECTIVE: To assess performance of a synthetic carbohydrate antigen for use in an enzyme immunoassay for diagnosis of trichinellosis in swine. ANIMALS AND SAMPLE POPULATION: 47 York X Duroc pigs and field sera from 265 farm pigs raised under various management systems. PROCEDURES: Each of 47 pigs was inoculated with 25 to 500 Trichinella spiralis larvae, and blood samples were obtained weekly. At postinoculation week 12, pigs were euthanatized, and tissues recovered from the tongue and diaphragm were digested to determine worm burden. Serum samples from experimentally infected pigs and sera obtained from pigs on a farm were tested by enzyme immunoassay, using standard excretory-secretory and synthetic glycan antigens. RESULTS: Of the 47 pigs, 46 (97.8%), with worm burden ranging from 0.02 to 248.8 larvae/g of tissue in the tongue and diaphragm, tested seropositive using both antigens. Time of seroconversion varied among pigs and was negatively correlated with intensity of infection. Minor differences were observed in time of seroconversion between the 2 antigens in 11 of 46 pigs, suggesting some differences in the antibody response. One pig with a worm burden of 0.01 larva/g was not detected by enzyme immunoassay using either antigen. Background values obtained using the 2 antigens did not differ among confinement-raised pigs, but background values for pigs raised outdoors on dirt lots were significantly lower using the glycan antigen Low-level, naturally acquired T spiralis infections in pigs were detected in most instances by use of both antigens, although there were some differences in antibody responses of infected pigs. CONCLUSION: The synthetic glycan antigen has potential for replacing excretory-secretory antigens as a standardized reagent for diagnosis of trichinellosis in pigs. PMID- 9401692 TI - Differences between longitudinal and circular smooth muscle in beta-adrenergic control of motility of isolated equine ileum. AB - OBJECTIVE: To identify beta-adrenoceptor subtypes involved in motility inhibition of circular and longitudinal smooth muscle layers of equine ileum. SAMPLE POPULATION: Isolated strips of equine ileum circular smooth muscle and membrane preparations from circular and longitudinal muscle layers. PROCEDURE: Functional assays of circular muscle preparations and radioligand binding assays and measurements of cAMP production in smooth muscle membranes from circular and longitudinal layers. RESULTS: Selective beta-adrenergic agonists exerted inhibitory effects on circular muscle preparations. Binding studies of cell membranes indicated that the density and distribution of 3 beta-adrenoceptor subtypes did not differ between longitudinal and circular muscle layers. Measurement of cAMP production in membrane preparations of longitudinal and circular muscle after selective beta-stimulation confirmed presence of the 3 adenylate cyclase-coupled beta-adrenoceptor subtypes; however, preparations from the 2 layers had differing cAMP production efficacy. CONCLUSIONS: The data may partly explain the differing functional responses between circular and longitudinal muscle preparations. CLINICAL RELEVANCE: Findings support the important role of beta-atypical adrenoceptors in the inhibitory regulation of equine ileum motility. PMID- 9401694 TI - Insulin-like growth factor I peptide elution profiles from fibrin polymers determined by use of high-performance liquid chromatography. AB - OBJECTIVE: To develop a high-performance liquid chromatography (HPLC) assay for insulin-like growth factor I (IGF-I) and to use it to quantify elution of IGF-I from polymerized fibrin in an in vitro system. SAMPLE POPULATION: Equine fibrinogen and calcium-activated bovine thrombin were used to form fibrin containing human recombinant IGF-I. PROCEDURE: Multiple fibrin disks were formed from polymerized fibrinogen and thrombin; 6 disks were loaded with 25 micrograms of recombinant human IGF-I at the time of polymerization, and 4 remained as unladen controls. The resultant clots were incubated at 37 C and 90% humidity for 22 days. Phosphate-buffered saline solution in each well was replaced daily, and IGF-I content was assayed by HPLC. Solid-phase separation was used to assay free IGF-I peptide peaks. A scan was done to determine optimal wavelength for IGF-I absorbance. Commercially pure IGF-I was used to construct a standard curve, and the IGF-I content of medium removed from all 10 wells each day was assayed. RESULTS: Pure unbound IGF-I eluted from the fibrin polymers for 22 days; initial rapid daily release of 1.6 to 1.7 micrograms of IGF/ml of medium changed after day 3 to commence an asymptotic decrease to 110 ng of IGF/ml by day 22. The fibrin disks had dissolved by day 22, and the experiment was terminated. Control disks did not have detectable IGF-I content at any time. Limit of the HPLC assay was 25 ng of IGF-I/ml. Retention time for nonprotein-bound IGF-I was 10.3 +/- 0.15 minutes. CONCLUSION: IGF-I (25 micrograms) can be added to polymerized fibrin and eluted as free ligand over a 22-day period of culture at 37 C. Release of IGF-I was initially independent of fibrin dissolution, but later appeared to follow a pattern consistent with fibrin degradation. CLINICAL RELEVANCE: IGF-I can be incorporated as a depot form in polymerized fibrin and is released over time in sufficient concentration to effectively stimulate articular chondrocyte metabolic activity. PMID- 9401693 TI - Pharmacokinetics of cisapride in horses after intravenous and rectal administration. AB - OBJECTIVES: To determine the i.v. pharmacokinetics of cisapride and measure systemic absorption after rectal administration. ANIMALS: 5 healthy adult mares (380 to 610 kg). PROCEDURE: Cisapride was administered, i.v., at a dosage of 0.1 mg/kg of body weight. In the same horses, after a 1-week washout period, cisapride was administered rectally at a dosage of 1 mg/kg by mixing crushed tablets with propylene glycol and administering the mixture into the rectum. After each drug administration, a series of blood samples were collected. Plasma was obtained and analyzed by high-performance liquid chromatography to determine cisapride concentration profiles after each drug administration. RESULTS: After i.v. administration, peak plasma concentration was 221.4 ng/ml and harmonic mean half-life was 1.9 hours. Rectal absorption of cisapride was negligible. Cisapride was detected in plasma from only 3 of 5 horses for which mean systemic availability was 1.23%. Mean maximal plasma concentration after rectal administration of cisapride was 13.5 ng/ml. CONCLUSION AND CLINICAL RELEVANCE: After i.v. administration of cisapride, plasma concentration is high for approximately 2 hours. Cisapride mixed with propylene glycol and administered rectally at a dosage of 1 mg/kg is poorly and incompletely absorbed. Thus, cisapride is not clinically useful for rectal administration in horses. PMID- 9401695 TI - Effect of low-dose atropine administration on dobutamine dose requirement in horses anesthetized with detomidine and halothane. AB - OBJECTIVE: To determine whether a low dose of atropine is associated with decreased requirement for cardiovascular supportive treatment in horses given detomidine prior to maintenance of general anesthesia with halothane. ANIMALS: 3 groups of 10 healthy horses. PROCEDURE: Detomidine (20 micrograms/kg of body weight, i.m.) was administered to all 30 horses. Then, 10 horses received atropine (0.006 mg/kg, i.v.) 1 hour after detomidine administration, 10 horses received atropine (0.012 mg/kg, i.m.) at the time of detomidine administration, and 10 horses served as a control group. Heart rate was measured prior to detomidine administration and at fixed intervals throughout anesthesia. The dobutamine infusion rate necessary to maintain mean arterial blood pressure between 70 and 80 mm of Hg was recorded. Systemic blood pressures, end-tidal halothane, end-tidal CO2, and arterial blood gas tensions were measured at fixed intervals. RESULTS: Mean heart rate was higher among horses receiving atropine i.v. or i.m., compared with that in control horses. Horses that received atropine i.v. had higher systemic arterial blood pressure and required a lower dobutamine infusion rate than did horses of the other groups. CONCLUSION: Detomidine treated, halothane-anesthetized horses given atropine i.v. required less dobutamine, compared with horses receiving or not receiving atropine i.m. Complications, such as colic and dysrhythmias, from use of higher doses of atropine, were not observed at this lower dose of atropine. CLINICAL RELEVANCE: i.v. administration of a low dose of atropine prior to induction of general anesthesia may result in improved blood pressure in horses that have received detomidine before anesthesia with halothane. PMID- 9401696 TI - Effects of medetomidine on serum insulin and plasma glucose concentrations in clinically normal dogs. AB - OBJECTIVE: To determine the effects of medetomidine, administered i.v., on serum insulin and plasma glucose concentrations in clinically normal dogs. ANIMALS: 6 healthy adult Beagles. PROCEDURE: Dogs were randomly assigned to each of 3 treatments in a prospective cross-over study design. Serum insulin and plasma glucose concentrations were determined before and 20, 40, 60, 120, 180, 240, 300, 360, 420, and 480 minutes after i.v. administration of 0.9% NaCl solution (control) or medetomidine (10 or 20 micrograms/kg of body weight). RESULTS: Mean serum insulin concentration decreased after medetomidine administration and was significantly (P < or = 0.05) lower than control values 20, 40, 60, and 120 minutes after drug administration. Mean plasma glucose concentration tended to increase after medetomidine administration, but did not differ significantly from control values. CONCLUSIONS: In dogs, i.v. administration of medetomidine at dosages of 10 and 20 micrograms/kg transiently decreases serum insulin concentration, but plasma glucose concentration remains within the normal physiologic range. CLINICAL RELEVANCE: Medetomidine can be given at low, preanesthetic dosages without significantly altering plasma glucose concentration in clinically normal dogs. PMID- 9401697 TI - Evaluation of selected cardiopulmonary and cerebral responses during medetomidine, propofol, and halothane anesthesia for laparoscopy in dogs. AB - OBJECTIVES: To compare the dose-sparing effect of medetomidine on the propofol induction dose and concentration of halothane for maintenance of anesthesia during laparoscopy and to provide guidelines for effective and safe use of these anesthetics in dogs to ensure desirable perioperative analgesia. ANIMALS: 14 purpose-bred dogs. PROCEDURE: Cardiopulmonary and electroencephalographic responses were determined during 2 anesthesia protocols in dogs scheduled for laparoscopy. Fifteen minutes before anesthesia induction, all dogs received atropine sulfate (0.02 mg/kg of body weight, i.m.). Seven dogs were then given propofol (6.6 mg/kg, i.v.); anesthesia was maintained with halothane in oxygen. The other dogs were given medetomidine hydrochloride (10 micrograms/kg, i.m.) 5 minutes after administration of atropine sulfate; anesthesia was then induced by administration of propofol (2.8 mg/kg, i.v.) and was maintained with halothane in oxygen. RESULTS: The halothane concentration required for laparoscopy was lower in dogs given medetomidine. Anesthetic requirements were significantly increased during abdominal manipulation in both groups. Total amplitude of the electroencephalograph in medetomidine-treated dogs was not significantly lower than that in dogs not given medetomidine. Pulmonary responses were stable throughout all procedures. The primary cardiovascular response was an increase in blood pressure associated with the medetomidine-atropine preanesthetic combination. Significant differences in total amplitude or frequency shifts (spectral edge) of brain wave activity were not associated with surgical stimulation. CONCLUSION: Lack of neurologic changes during laparoscopy supports the efficacy of either medetomidine-propofol-halothane or propofol-halothane combinations at higher concentrations to provide desirable analgesia and anesthesia in this group of dogs. PMID- 9401698 TI - Secondary stress responses to acute handling in striped bass (Morone saxatilis) and hybrid striped bass (Morone chrysops x Morone saxatilis). AB - OBJECTIVE: To test the hypothesis that, compared with pure striped bass, hybrid striped bass have reduced secondary physiologic responses to handling stress. A secondary objective was to determine whether not feeding fish for a 3-day period affected responses. ANIMALS: Hatchery-reared adult striped bass (Morone saxatilis) and adult hybrids of striped bass with white bass (M chrysops), with mean length of 27.9 cm and mean weight of 487 g. PROCEDURE: Fed and 3-day nonfed fish, in groups of 6, were held in dip nets above water for 3 minutes. Severity of response to handling was determined by measuring plasma glucose and chloride and blood lactic acid, sodium, and potassium concentrations. Terminal samples were taken from fish before handling (control), immediately after handling, and after 12, 24, and 48 hours of recovery. RESULTS: Striped bass were hyperglycemic and lactacidemic after stress and for 12 to 48 hours afterward, whereas glucose and lactic acid values in hybrids were essentially unchanged. Blood sodium and chloride concentrations of hybrids decreased after stress, then returned to control values within 24 hours. Striped bass, however, had a greater decrease in values for these electrolytes and failed to recover in 48 hours. Blood potassium concentration remained unchanged in all test groups. Nonfeeding for 3 days before handling did not appear to affect stress response in striped bass or hybrids. CONCLUSIONS AND CLINICAL RELEVANCE: Striped bass have an appreciably greater response to acute handling stresses, such as those that may be experienced in hatcheries and experimental laboratories, than do hybrid bass. Thus, pure striped bass require more care in handling. The usual practice of not feeding before handling does not affect physiologic responses. PMID- 9401699 TI - Metabolizable energy intake and sustained energy expenditure of Alaskan sled dogs during heavy exertion in the cold. AB - OBJECTIVE: To measure energy expenditures of Alaskan sled dogs at rest and during racing under frigid conditions, using the doubly labeled water (DLW) technique. ANIMALS: 18 fit Alaskan sled dogs. PROCEDURE: Energy expenditure was measured in 9 dogs during a 490-km sled dog race by use of the DLW technique, whereby dogs were administered water enriched with nonradioactive isotopes of hydrogen and oxygen. Energy intake was determined by dietary analysis. Changes in background abundance of the isotopes 2H and 18O were monitored in 5 dogs that did not receive isotope-enriched water. RESULTS: Dogs completed the 490-km race at an average speed of 7 km/h at ambient temperature of -35 to -10 C. Total energy expenditure, measured by the DLW technique, was 47,100 +/- 5,900 kJ/d (4,400 +/- 400 kJ.kg-0.75/d), and metabolizable energy intake was 44,600 kJ/d (4,100 kJ.kg 0.75/d) during the 70-hour race. CONCLUSIONS: The sustained metabolic rate for these sled dogs during racing was extraordinarily high for a large mammal. This study validated use of the DLW technique in dogs with exceptionally high energy expenditure associated with prolonged exercise in the cold. PMID- 9401700 TI - Comparison of alveolar ventilation, oxygenation, pressure support, and respiratory system resistance in response to noninvasive versus conventional mechanical ventilation in foals. AB - OBJECTIVE: To compare the efficacy of positive pressure ventilation applied through a mask versus an endotracheal tube, using anesthetized/paralyzed foals as a model for foals with hypoventilation. ANIMALS: Six 1-month-old foals. PROCEDURE: A crossover design was used to compare the physiologic response of foals to 2 ventilatory techniques, noninvasive mask mechanical ventilation (NIMV) versus endotracheal mechanical ventilation (ETMV), during a single period of anesthesia and paralysis. Arterial pH, PaO2, PaCO2, oxygen saturation, end-tidal CO2 tension, airway pressures, total respiratory system resistance, resistance across the upper airways (proximal to the midtracheal region), and positive end expiratory pressures (PEEP) were measured. Only tidal volume (VT; 10, 12.5, and 15 ml/kg of body weight) or PEEP (7 cm of H2O) varied. RESULTS: Compared with ETMV, use of NIMV at equivalent VT resulted in PaCO2 and pH values that were significantly higher, but PaO2 was only slightly lower. Between the 2 methods, peak airway pressure was similar, but peak expiratory flow was significantly lower and total respiratory resistance higher at each VT for NIMV. Delivery of PEEP (7 cm of H2O) was slightly better for ETMV (7.1 +/- 1.3 cm of H2O) than for NIMV (5.6 +/- 0.6 cm of H2O). CONCLUSION: These data suggest that use of NIMV induces similar physiologic effects as ETMV, but the nasal cavities and mask contribute greater dead space, manifesting in hypercapnia. Increasing the VT used on a per kilogram of body weight basis, or the use of pressure-cycled ventilation might reduce hypercapnia during NIMV. CLINICAL RELEVANCE: Use of NIMV might be applicable in selected foals, such as those with hypoventilation and minimal changes in lung compliance, during weaning from endotracheal mechanical ventilation, or for short-term ventilation in weak foals. PMID- 9401701 TI - Exercise capacity in young and old mares. AB - OBJECTIVE: To test the hypothesis that, compared with unfit young horses, unfit older horses have lower aerobic capacity and reduction in other indices of exercise capacity. ANIMALS: 6 young (mean +/- SEM, 5.3 +/- 0.8 years and 445 +/- 13 kg) and 6 aged (22.0 +/- 0.4 years and 473 +/- 18 kg) healthy Standardbred and Thoroughbred mares. PROCEDURES: The mares, accustomed to running on a treadmill, were tested by use of an incremental exercise test. None of the mares had received exercise training for at least 4 months prior to the study. During testing, mares ran up a fixed 6% grade, starting at a speed of 4 m/s, with 1 m/s increase every 60 seconds (omitting 5 m/s) until they reached fatigue. Maximal oxygen uptake (VO2max) was measured by use of an open-flow calorimeter. Venous blood samples (10 ml) were collected during the last 10 seconds of each step and were used to measure blood lactate concentration and PCV. Calculated performance indices included velocity at VO2max, maximal velocity, and velocity at lactate concentration of 4 mmol/L; work rate (watts) at those velocities also was determined. RESULTS: There were differences (P < 0.05) between old and young mares for maximal run velocity attained during the test (8.7 +/- 0.5 versus 10.8 +/- 0.5 m/s, respectively), VO2max (89.4 +/- 4.3 versus 117.3 +/- 9.5 ml/kg of body weight/min, respectively), and velocity at VO2max (8.0 +/- 0.4 versus 9.8 +/ 0.7 m/s, respectively). Also, velocity required to reach blood lactate concentration of 4 mmol/L was lower (P < 0.05) in old (7.5 +/- 0.4 m/s), compared with young (10.2 +/- 0.7 m/s), mares. CONCLUSION: Older mares have substantially (-24%) lower maximal aerobic capacity than do young mares. CLINICAL RELEVANCE: Many horses participate in athletic activities into their late teens and some do so beyond the age of 20 years; thus, the need exists to explore ways to adjust training programs for older horses. PMID- 9401703 TI - The problems of inattention: methods and interpretations. PMID- 9401702 TI - Rehabilitation of dogs with surgically treated cranial cruciate ligament deficient stifles by use of electrical stimulation of muscles. AB - OBJECTIVE: To determine effect of electrical muscle stimulation (EMS) on rate and degree of return to function of the limb and development of degenerative joint disease (DJD) after surgical creation and subsequent stabilization of the cranial cruciate ligament (CrCL)-deficient stifle. ANIMALS: 12 clinically normal adult large (19.5 to 31.5 kg) dogs. PROCEDURE: Dogs were anesthetized, and the right CrCL was severed via arthrotomy, destabilizing the stifle. After 3 weeks, the stifle was surgically stabilized. Three weeks later, 6 dogs were subjected to an EMS treatment protocol for the thigh muscles. At 5, 9, 13, and 19 weeks after stifle destabilization, treated (n = 6) and control (n = 6) dogs were evaluated for return of stifle function. Gross and histologic evaluations of the stifles were performed at 19 weeks after stifle destabilization. RESULTS: Treated dogs had significantly (P = 0.001) better lameness score than did control dogs. There was less palpable crepitation of the stifle in treated dogs (P = 0.06); treated dogs also had significantly (P = 0.01) fewer radiographic signs of bone changes. Thigh circumference was significantly (P = 0.02) larger in treated dogs. There was less gross cartilage damage (P = 0.07) in the EMS-treated dogs, but more medial meniscal damage (P = 0.058, cranial pole; P = 0.051, caudal pole). CONCLUSIONS: Improved lameness scores, larger thigh circumference, and decreased radiographically apparent bony changes observed for the treated group of dogs support the hypothesis that dogs treated by EMS after surgical stabilization of the CrCL-deficient stifle had improved limb function, with less DJD, than did dogs treated with the currently accepted clinical protocol of cage rest and slow return to normal activity. However, results of force plate evaluation did not support the hypothesis. Increased meniscal damage in dogs treated by EMS may be cause for concern. PMID- 9401704 TI - The switching model of latent inhibition: an update of neural substrates. AB - Organisms exposed to a stimulus which has no significant consequences, show subsequently latent inhibition (LI), namely, retarded conditioning to this stimulus. LI is considered to index the capacity to ignore irrelevant stimuli and its disruption has recently received increasing interest as an animal model of cognitive deficits in schizophrenia. Initial studies indicated that LI is disrupted by systemic or intra-accumbens injections of amphetamine and hippocampal lesions, and potentiated by systemic administration of neuroleptics. On the basis of these findings, the switching model of LI proposed that LI depends on the subicular input to the nucleus accumbens (NAC). Subsequent studies supported and refined this proposition. Lesion studies show that LI is indeed disrupted by severing the subicular input to the NAC, and further implicate the entorhinal/ventral subicular portion of this pathway projecting to the shell subterritory of the NAC. There is a functional dissociation between the shell and core subterritories of the NAC, with lesions of the former but not of the latter disrupting LI. This suggests that the shell is necessary for the expression and the core for the disruption of LI. The involvement of the NAC has been also demonstrated by findings that LI is disrupted by intra-accumbens injection of amphetamine and potentiated by DA depletion or blockade in this structure. Disruption and potentiation of LI by systemic administration of amphetamine and neuroleptics, respectively, have been firmly established, and in addition, have been shown to be sensitive to parametric manipulations of the LI procedure. LI is unaffected by lesions and DA manipulations of medial prefrontal cortex and lesions of basolateral amygdala. The implications of these findings for LI as an animal model of schizophrenia are discussed. PMID- 9401705 TI - Latent inhibition: the nucleus accumbens connection revisited. AB - It has been proposed that dopaminergic transmission in the nucleus accumbens plays a key role in regulating latent inhibition (LI), i.e. the retardation of conditioning that occurs if a to-be-conditioned stimulus is first presented a number of times ('preexposure') without other consequence. New evidence in support of this hypothesis is presented or reviewed here, showing that: (1) intra accumbens injection of haloperidol at the time of conditioning potentiates LI; (2) destruction of dopaminergic terminals in the nucleus accumbens potentiates LI; (3) intra-accumbens haloperidol reverses the blockade of LI caused by systemic nicotine; (4) intra-accumbens haloperidol reverses the blockade of LI caused by systemic amphetamine; (5) after a single systemic injection of amphetamine (insufficient on its own to block LI), a subsequent intra-accumbens injection of amphetamine at the time of conditioning blocks LI; and (6) intra accumbens (like systemic) amphetamine administered 15 min before conditioning, without prior systemic amphetamine, failed to block LI. The difference between the effects on LI of one and two administrations of amphetamine, respectively, is interpreted in terms of the need for sensitisation of the response to amphetamine, with the result that the response to the second administration includes a component of impulse-dependent dopamine release in the nucleus accumbens that is otherwise lacking. Data from dialysis experiments suggest that such impulse-dependent accumbens dopamine release also occurs at relatively long delays after a single systemic administration of amphetamine. It was accordingly predicted, and found, that, although LI is intact 15 min after an i.p. injection (confirming previous results), it is abolished at 90 min after the injection of amphetamine. This finding is consistent with the effects of amphetamine in human subjects, in whom LI is blocked 90 min after a single oral administration. Overall, these results strengthen the case that the blockade of LI by elevated, and potentiation of LI by decreased, dopaminergic transmission are both due specifically to actions in the nucleus accumbens; and also add to the similarities between LI studied in animal and human subjects, respectively. PMID- 9401706 TI - Amphetamine-induced disruption and haloperidol-induced potentiation of latent inhibition depend on the nature of the stimulus. AB - If a stimulus (e.g. light) is repeatedly preexposed without consequences, it subsequently develops a weaker association with a reinforcer (e.g. foot shock) than does a non-preexposed stimulus. This retarded conditioning to the preexposed as compared to the non-preexposed stimulus, is latent inhibition (LI). It is well documented that LI is disrupted by low doses of amphetamine and potentiated by neuroleptic drugs, and there is evidence that the action of these agents on LI can be modified by changes in the parameters of preexposure or conditioning. The present experiments tested whether the effects of DA agents on LI are influenced by the nature of the stimulus. In two experiments, LI was assessed using an off baseline conditioned emotional response (CER) procedure in rats licking for water, consisting of three stages: preexposure, in which the stimulus (a light) to be conditioned, was repeatedly presented without being followed by reinforcement; conditioning, in which the preexposed stimulus was paired with reinforcement (a foot-shock); and test, in which LI was indexed by animals' degree of suppression of licking during stimulus presentation. In both experiments, different groups of animals were preexposed and conditioned with four different preexposed visual stimuli: three steady side-lights, three flashing side-lights, one flashing side-light, and a flashing houselight. Experiment 1 used 40 stimulus preexposures and tested the effects of 1 mg/kg D amphetamine, whereas experiment 2 used 10 preexposures and tested the effects of 0.1 mg/kg haloperidol. The results showed that of the four stimuli used, both drugs were effective with only one and the same stimulus, namely, flashing houselight. This demonstrates that the disruptive effect of amphetamine and the potentiating effect of haloperidol on LI, are modifiable by manipulating the nature of the preexposed stimulus. PMID- 9401707 TI - Disruption of latent inhibition in the rat by the 5-HT2 agonist DOI: effects of MDL 100,907, clozapine, risperidone and haloperidol. AB - Latent inhibition (LI), a measure of the ability to learn to ignore irrelevant stimuli, is disrupted in acute schizophrenics and in rats treated with amphetamine; antipsychotics prevent amphetamine disruption of LI in rats. The 5 HT2A/C agonist 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI) has hallucinogenic properties in humans, and evidence suggests that 5-HT2 antagonism is an important component of atypical antipsychotic activity. Therefore, the ability of DOI to disrupt LI in rats was tested, and the ability of clinically used and putative antipsychotics to reverse DOI disruption of LI was assessed. The method consisted of four phases. After habituation to the apparatus, thirsty rats underwent preexposure to a tone stimulus 24 h prior to two tone-shock conditioning trials. LI was demonstrated at testing (an additional 24 h later) by reduced lick suppression during tone presentation. When administered at the preexposure phase only, DOI disrupted LI. However, when administered at both preexposure and conditioning phases, DOI did not disrupt LI except at the highest dose, where lick suppression itself was also disrupted. Therefore, disruptive effects of DOI on LI are not easily dissociated from state-dependent learning effects. Additional experiments demonstrated that haloperidol, clozapine, risperidone, and the selective 5-HT2A antagonist MDL 100,907 prevented the disruptive effects of DOI on LI when administered at preexposure only. These results agree with findings that these compounds can also prevent other behavioral effects of DOI. Further experiments will be required to explore the possible involvement of state-dependent learning effects in the present results. However, if the disruptive effects of DOI on LI are due to an influence on attentional processes rather than state-dependent learning, this procedure may have potential as a method for detection of antipsychotic activity. PMID- 9401708 TI - WAY100635 and latent inhibition in the rat: selective effects at preexposure. AB - The influence of the selective, silent 5HT1a antagonist WAY100635 (Wyeth Research Ltd) on the latent inhibition effect was examined in a within-subject, on baseline conditioned suppression procedure in rats. WAY100635 was found to enhance the latent inhibition effect, producing a retardation in the acquisition of conditioned suppression following a level of stimulus preexposure known to be insufficient to produce a latent inhibition effect in control animals. This influence of the drug was restricted to its actions during the preexposure phase of the experiment, and the drug also abolished the unconditioned suppression of lever pressing that occurs on the first presentation of a novel auditory stimulus. These findings are discussed in terms of the possible influence of serotonergic manipulations on contextual processing, and also have important implications for current animal models of schizophrenia which stress the role of dopaminergic mechanisms in latent inhibition. PMID- 9401709 TI - Bilateral ablation of the auditory cortex in the rat alters conditioned emotional suppression to a sound as appraised through a latent inhibition study. AB - Latent inhibition consists of a retardation of conditioning seen when the to be conditioned stimulus is presented a number of times with no other consequence. This phenomenon likely reflects processes of selective attention whereby irrelevant stimuli come to be ignored. Using physiological models for auditory attention, some investigators have suggested that selective attention acts as a filtering mechanism capable of inhibiting or gating unattended stimuli relative to attended ones in the auditory cortex. In the present work, an on-baseline conditioned suppression response procedure was used to study the effects of stimulus preexposure in rats submitted to bilateral auditory cortex ablation. Our results indicate that both auditory cortex lesioned and control animals exhibit latent inhibition to a sound. However, learning after preexposure to that sound was particularly slow in animals with bilateral auditory cortex lesion, i.e. in these animals, the latent inhibition effect appeared to be enhanced. Conditioning from one day to the next also varied slightly. Thus, the auditory cortex appears to modulate learning when the conditioned stimulus is a sound. PMID- 9401710 TI - Fetal hippocampal transplants restore conditioned blocking in rats with dorsal hippocampal lesions: effect of age. AB - Previous studies have shown that conditioned blocking of taste aversion learning in 3-month-old Wistar rats depends on the hippocampal system integrity. Thus, the aim was to demonstrate that enough connectivity would develop after a graft to support the attention mechanism required for conditioned blocking. In the first experiment, bilateral homotopic grafts of 16-17 day-old hippocampal fetal tissue applied to 3-month-old male Wistar rats after electrolytical lesions of the dorsal hippocampus reinstated conditioned blocking tested 5 months after the transplantation. Unexpectedly, an early age-dependent impairment of conditioned blocking, similar to that induced by hippocampal lesions, was found in the 8 month-old control group. This finding was further supported by the results of the second experiment. Non-operated 3-month-old but not 8-month-old Wistar rats showed conditioned blocking. The results are discussed in terms of early hippocampal vulnerability, prevented by fetal grafts. PMID- 9401711 TI - Latent inhibition as a measure of learned inattention: some problems and solutions. AB - The latent inhibition (LI) paradigm has been used to assess attentional dysfunction in various pathological groups. The rationale is based on the assumption that passive stimulus exposure results in the acquisition of an inattentional response to that stimulus. Consequently, compared to a novel stimulus in the same new learning situation, the preexposed stimulus is at a disadvantage. It is argued that methodological and conceptual problems in constructing procedures and designs have created obstacles in relating disrupted LI to psychopathology. Specifically, issues associated with within- and between subject designs, dichotomous dependent variables, ceiling effects, converging operations, and possible mis-attribution of the LI effect are addressed. Designs and data from several new human-LI paradigms, with normal, de novo Parkinson, and schizophrenic subjects are examined. Results from a multi-condition, within subject visual search procedure suggest that LI, heretofore attributed only to a deficit in the stimulus preexposed group, may, in part, be due to enhanced performance in the nonpreexposed group. Implications for the design and interpretation of LI experiments, particularly with pathologic groups are discussed. PMID- 9401712 TI - Latent inhibition and autonomic responses: a psychophysiological approach. AB - Latent inhibition, retarded learning after preexposure to the to-be-conditioned stimulus, has been implied as a tool for the investigation of attentional deficits in schizophrenia and related disorders. The present paper reviews research that used Pavlovian conditioning as indexed by autonomic responses (electrodermal, vasomotor, cardiac) to investigate latent inhibition in adult humans. Latent inhibition has been demonstrated repeatedly in healthy subjects in absence of a masking task that is required in other latent inhibition paradigms. Moreover, latent inhibition of Pavlovian conditioning is stimulus-specific and increases with an increased number of preexposure trials which mirrors results from research in animals. A reduction of latent inhibition has been shown in healthy subjects who score high on questionnaire measures of psychosis proneness and in unmedicated schizophrenic patients. The latter result was obtained in a within-subject paradigm that holds promise for research with patient samples. PMID- 9401713 TI - The development of conditioned blocking and monoamine metabolism in children with attention-deficit-hyperactivity disorder or complex tics and healthy controls: an exploratory analysis. AB - Conditioned blocking (CB) measures the transient suppression of learning that a new stimulus, added during learning, has the same consequences as the conditioned stimulus already present. Normal CB increases between the age of 8 and 20 years (Oades, R.D., Roepcke, B. and Schepker, R., A. test of conditioned blocking and its development in childhood and adolescence: relationship to personality and monoamine metabolism, Dev. Neuropsychol., 12 (1996) 207-230). In the present study CB development is compared between healthy children (CN), children with attention deficit (ADHD) and those with complex tics or Tourette's syndrome (TS) with mean ages of 10-11 years. All children needed fewer learning trials with increasing age: the ADHD group showed a slight impairment. Only controls improved CB with increasing age. A trend for worse CB in the TS than the other groups was significant for those over 11 years. While ADHD children over 11 years showed less CB than controls, younger ADHD children showed more. A correlational analysis of the status of monoamine metabolism in 24 h urine samples showed a positive relationship for CB with dopamine metabolism in controls and TS children, but a negative relationship in ADHD children. In contrast, increases of serotonin metabolism were negatively related to CB in TS but positively in ADHD patients. In conclusion, when selective information processing abilities reflected by CB start to develop at puberty-onset, there is a relative worsening in ADHD patients. But TS patients show an impairment independent of age. Changes in the balance between dopamine and serotonin systems may contribute to normal and abnormal cognitive development. PMID- 9401714 TI - Disruption of the Kamin blocking effect in schizophrenia and in normal subjects following amphetamine. AB - The Kamin blocking effect (KBE) is an established animal learning paradigm measuring selective processing, in which reduced blocking reflects allocation of greater processing resources to non-relevant information. Two KBE tasks are described below. Results from studies using the first (between-subjects) task indicate that KBE is abolished in acute schizophrenics with positive psychotic symptoms. It is also abolished in the relatives of schizophrenic subjects, although interpretation of this finding is hampered by poor performance of subjects in the control condition. The second (within-subjects) task indicated abolition of KBE in schizophrenic patients with positive psychotic symptoms. Administration of acute amphetamine to normal human subjects did not significantly disrupt performance on the first task. Whilst for the second task, although blocking was limited to placebo subjects, overall pre-exposure effects are not sufficiently strong to indicate specific drug effects. PMID- 9401715 TI - Stimulus dimension shifts in patients with schizophrenia, with and without paranoid hallucinatory symptoms, or obsessive compulsive disorder: strategies, blocking and monoamine status. AB - Reversal, and intra-dimensional (ID) and extra-dimensional (ED) nonreversal discrimination shifts were studied to see if learned inattention to the irrelevant dimension differentially influenced the efficacy of learning and stimulus choice strategy. Performance was compared with conditioned blocking (CB) and monoamine metabolic status between healthy controls, patients with obsessive compulsive disorder (OCD) or schizophrenia with (PH) or without (NP) active paranoid hallucinatory symptoms. PH and NP patients improved learning with practice, but showed an impaired shift on each task. OCD patients were impaired only on the ED-shift. The NP patient's impairment was nonspecific and, unlike PH and controls, it related to reversal performance. All subjects acquired an attentional set for colour reflected in the length of stimulus-response sequences. Analysis of paired-stimulus choice-strategies showed that while all patients showed fewer correct win-stay choices, only PH patients perseverated with lose-stay choices. Learning about the added stimulus in the CB task related to ID-shift efficiency in NP patients. Increases of dopamine activity related to delayed learning but more switches of stimulus choice in the shift-tasks. Increases of serotonin activity correlated with faster learning in controls, OCD and PH patients. In NP patients the opposite held for dopamine and serotonin activity. Thus the two learned inattention tasks have different if related requirements and correlates: the data are consistent with the use of automatic exogenous attention strategies by NP patients, of inefficient controlled attention by PH patients and the automatization of endogenous processes in controls. PMID- 9401716 TI - Species differences in cardiac thyroid hormone receptor isoforms protein abundance. AB - Little is known about the cardiac expression of different thyroid hormone receptor (TR) isoforms. The aim of the study was to investigate such patterns of TR expression at the protein level in different species and in some human tissues. Western blot analysis with specific polyclonal rabbit antibodies to each TR isoform was performed with samples from myocardium of the left ventricle from man, dog, guinea pig, rat and mouse, as well as with samples from several human tissues such as heart, skeletal muscle, brain, liver and thyroid. The TR alpha 1 isoform was present in all of the species examined. The TR alpha 2 was recognized in human, dog and guinea pig heart, while no such band was recognized in rat and mouse hearts. TR beta 1 was not detected in the human heart but in the other species. Similarly to TR alpha 1, TR beta 2 was detected in all of the species examined. In the human tissues studied, TR alpha 1 was detected in heart and skeletal muscle, whereas TR alpha 2 was found only in the heart. TR beta 1 was not detected in any of the examined human tissues, while TR beta 2 was found in all of them. These results revealed unique distributions of TR variants and they demonstrate common epitopes in TR in the different species. For the first time, the presence of a TR beta 2 isoform has been shown in human tissues. TR isoforms may have a tissue and species specific role in the regulation of gene expression and may in part explain variable tissue effects of thyroid hormones. PMID- 9401717 TI - Cloning of the cDNA for a mouse homologue of human PHBP: a novel hyaluronan binding protein. AB - The cDNA which encodes the mouse counterpart of human plasma hyaluronan-binding protein (PHBP) was isolated and characterized. The clone contained an insert of 2153 bp, which contained the 1674-bp open reading frame coding for a polypeptide of 558 amino acid residues. The amino acid sequence of mouse PHBP predicted from the nucleotide sequence of cDNA shows reasonable homology to that of human PHBP. Like human PHBP, the amino acid sequence predicted from the nucleotide sequence of mouse PHBP cDNA exhibited significant homology to that of human hepatocyte growth factor activator (HGFA). PMID- 9401718 TI - Inhibitory effect of oversulfated fucoidan on tube formation by human vascular endothelial cells. AB - Fucoidan is a sulfated poly(L-fucopyranose) present in brown marine algae. In this study, we examined the effect of native and chemically oversulfated fucoidans (NF and OSF) on the tube structure formation by human umbilical vein endothelial cells (HUVEC) on the basement membrane preparation, Matrigel. Unlike NF, OSF significantly decreased the tube formation: maximal inhibition (50% of control) was obtained with 25 micrograms/ml. The OSF effect was mediated, at least in part, through the inhibition of HUVEC migration, as determined by the ability to block chemotaxis in a Transwell chamber assay. Quantitative immunoreactive assays for tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor-1 (PAI-1) in the culture media indicated that OSF (25 micrograms/ml) increased the accumulation of PAI-1 antigen, but not of t-PA antigen, 2.7-fold compared with control. The release of both antigens by HUVEC was slightly affected by the addition of NF. Determination of the media levels of type IV collagenase activity and tissue inhibitor of metalloproteinase-1 (TIMP-1) antigen showed that OSF (25 micrograms/ml) decreased the collagenolytic activity by 50% compared to the control, without alteration of the TIMP antigen level. However, the collagenase inhibition by OSF was not observed in an assay system using purified enzyme. NF had no effect on collagenase activity or TIMP-1 antigen levels. These results indicate that the introduction of sulfate groups into NF enables it to effectively inhibit the formation of capillary-like structures by HUVEC on Matrigel by reducing the basement membrane destruction and cell migration. It is involved as at least one of the mechanisms by which the OSF induced increase in HUVEC PAI-1 decreases plasmin formation and suppresses the following pro-collagenase activation. PMID- 9401719 TI - Inhibition of the metallo-beta-lactamase produced from Serratia marcescens by thiol compounds. AB - Low molecular weight thiol compounds have been found to be strong inhibitors of metallo-beta-lactamase (IMP-1) produced by Serratia marcescens TN9106, which was expressed by Echerichia coli JM109 cells. Mercaptoacetic acid and 2 mercaptopropionic acid strongly and competitively inhibited IMP-1 with Ki of 0.23 and 0.19 microM, respectively. 2-Mercaptoethanol reversibly inhibited IMP-1 but did not show simple competitive inhibition. PMID- 9401720 TI - Separation of sialoglycoprotein fractions having influenza virus-receptor activity from human meconium and some of their chemical properties. AB - Three sialoglycoprotein fractions having reactivities against influenza A and B viruses (IV-A and IV-B) and influenza virus-hemagglutinin (IV-HA), were obtained by gel filtration using a Sephacryl S-300 column and an Asahipak GS-710 HPLC column from water-soluble fraction prepared from human meconium (ME-WSF). Molecular weights of sialoglycoproteins in these fractions were estimated as 746 kDa (P-Fr. 1-3), 4470 kDa (P-Fr. 1-2) and a more than 10,000 kDa (P-Fr. 1-1), respectively. The other in P-Fr. 2, with molecular weight estimated as 92 kDa, revealed positive but weak reactivity against IV-HA; however, the fraction did not show positive reactivity against either IVs-A or -B. All these fractions contained sialoglycoproteins with a high content of carbohydrates (31.0-59.3%, w/w). From their carbohydrate compositions, it can be suspected that sialoglycoproteins having O-glycosidically linked carbohydrate chains were predominating in the three fractions. In conclusion, it is suggested that at least three sialoglycoproteins with influenza virus-receptor activity showing different chemical properties to each other exist in human meconium. PMID- 9401721 TI - The effect of TRK-530 on experimental arthritis in mice. AB - TRK-530 is a newly synthesized diphosphonate derivative. We investigated the effect of TRK-530 on type II collagen-induced arthritis (CIA) in mice in comparison to that of prednisolone and indomethacin. TRK-530 at a dose of 25 mg/kg showed a tendency to inhibit CIA. TRK-530 at a dose of 50 mg/kg inhibited the development of the CIA in terms of the progression of footpad swelling, bone damage and histopathological changes. TRK-530 at a dose of 50 mg/kg also significantly inhibited the delayed type hypersensitivity (DTH) response to type II collagen, but not the production of anti-type II collagen IgG antibody in arthritic mice. To investigate the inhibitory mechanism of TRK-530, the type of effect of TRK-530 on the production of IL-1 beta in vitro was studied. TRK-530 at a concentration of 10(-4) M inhibited LPS-induced IL-1 beta production from J774.1 cells. In conclusion, TRK-530 inhibited CIA in mice. The inhibition of the DTH reaction to type II collagen and the inhibition of IL-1 beta production may partly participate the anti-rheumatoid action of TRK-530. PMID- 9401722 TI - The analgesic and anti-inflammatory effects of shark cartilage are due to a peptide molecule and are nitric oxide (NO) system dependent. AB - The present work shows an antinociceptive and dose-dependent effect of shark cartilage hydrosoluble fraction (HF) on writhing and formalin tests in mice. The effect was not altered by thalidomide, a known inhibitor of tumor necrosis factor alfa (TNF-alfa) synthesis. Similarly, the antinociceptive effect did not change in the presence of naloxone, indicating that the opioid system is not involved. However, the effect observed was blocked by L-arginine, a NO synthesis substrate, and it was potentiated by L-NAME, suggesting a role of the NO system in the shark cartilage antinociceptive effect. Effects similar to those seen with the HF were detected with peak II from gel filtration chromatography. The increase in vascular permeability induced by serotonin in rats was significantly abolished by the HF at the dose of 2 mg/kg, p.o., and again it was not potentiated by thalidomide. The observed blockade in the vascular permeability increase induced by histamine was detected only with a higher dose (10 mg/kg, p.o.). PMID- 9401723 TI - The effects of hange-shashin-to on gastric function in comparison with sho-saiko to. AB - The effects of "Hange-shashin-to (TJ-14)" on gastric function were examined in comparison with "Sho-saiko-to (TJ-9)". Oral treatment with TJ-14 (125-500 mg/kg) caused dose-dependent suppression of ethanol-induced gastric injury, while it did not suppress gastric lesions induced by water-immersion stress. TJ-9 (125-500 mg/kg, p.o.) suppressed both water-immersion stress-induced gastric lesions and ethanol-induced gastric injury in a dose-dependent manner. Intraduodenal administration of TJ-14 even at 500 mg/kg did not affect gastric juice secretion, while TJ-9 at 125 to 500 mg/kg dose-dependently suppressed gastric juice secretion. TJ-14 (125-500 mg/kg, p.o.) accelerated gastric emptying in normal rats and improved the delayed gastric emptying induced by BaCl2 in a dose dependent manner, whereas such effect was not noted with TJ-9. Oral treatment with TJ-14 at 500 mg/kg significantly suppressed apomorphine-induced vomiting, but it did not affect copper sulfate-induced vomiting. These results suggest that TJ-14 exhibits an anti-ulcer action (probably based on its ability to protect the gastric mucosa), improvement of gastric emptying and an anti-emetic action. TJ-9 also showed anti-ulcer effects, probably based on its ability to suppress gastric secretion and to protect the gastric mucosa. Thus, the present study demonstrated the effectiveness of TJ-14 and TJ-9 against gastric disease, and provided basic data which explain the differences in clinical application between these two kampo medicines. PMID- 9401724 TI - Protective effect of tetrandrine on normal human mononuclear cells against ionizing irradiation. AB - Tetrandrine, an alkaloid isolated from the plant Stephania tetrandra, at low concentration (2 micrograms/ml) was shown to protect normal human mononuclear cells in vitro against damage due to a single high-dose of ionizing irradiation (10 Gy). The cell survival rate increased from 58.3 +/- 2.2% in the irradiated group to 78.0 +/- 2.6% in the tetrandrine-pretreated group, and similarly, the percentage of necrotic cells declined from 20.7 +/- 2.5% to 10.7 +/- 1.9%, respectively. This protective effect of tetrandrine for cell surviving fraction increased in a dose-dependent manner. Tetrandrine was also found to inhibit inflammatory responses induced by irradiation including the release of superoxide (NBT [nitroblue tetrazolium] reduction decreased from 21.3 +/- 2.3% to 10.2 +/- 2.5%) and phagocytic activity (decreased from 80.7 +/- 3.8% to 50.7 +/- 2.3%, the same range level as that of the control group). However, the alkaloid demonstrated no effect on the production of nitric oxide. In terms of cell morphology, only two types were observed-normal or necrotic cells, and there were no characteristics of programmed cell death. These results indicate that tetrandrine possesses radioprotective activity against 10 Gy of ionizing irradiation and could suppress irradiation-induced inflammatory processes. PMID- 9401725 TI - Studies on kochiae fructus. IV. Anti-allergic effects of 70% ethanol extract and its component, momordin Ic from dried fruits of Kochia scoparia L. AB - The 70% ethanol extract (KS-ext) from Kochiae Fructus (dried fruits of Kochia scoparia L.) has been screened for activity in experimental models of type I-IV allergy. In type I allergic models, KS-ext at doses of 200 and 500 mg/kg, p.o. exhibited an inhibitory effect on 48-h homologous passive cutaneous anaphylaxis (PCA) in rats, which is related to IgE, and 1.5-h heterologous PCA in mice, which is related to IgG. In a type III allergic model, KS-ext showed an inhibitory effect on direct passive arthus reaction (DPAR) in rats, while it had no inhibitory effect on reversed cutaneous anaphylaxis (RCA) in a type II allergic model. Furthermore, in a type IV allergic model, KS-ext had an inhibitory effect on the effector phase in picryl chloride-induced contact dermatitis (PC-CD). Also, its anti-pruritogenic component, momordin Ic (oleanane saponin) exhibited inhibitory effects on 48-h homologous PCA and PC-CD. These results indicate that Kochiae Fructus not only inhibits humoral immunity but also influences cellular immunity, and should be recognized as a material for anti-allergic reactions. Also, the mode of its anti-pruritogenic activity may be mediated by anti-allergic action, and its active component may be partially attributed to momordin Ic. PMID- 9401726 TI - Pharmacological properties of traditional medicines. XXIII. Searching for active compounds in the blood and bile of rats after oral administrations of extracts of sansohnin [symbol: see text]. AB - We made a trial of searching for the bioactive substances from Sansohnin [symbol: see text]. The blood and bile of rats after the oral administration of extracts of Sansohnin were analyzed by three-dimensional high-performance liquid chromatography (3D-HPLC). In blood, spinosin and feruloyl spinosin were found after the oral administration of a butanol extract of Sansohnin. In bile, spinosin and feruloyl spinosin were also identified after the oral administration of a water extract of Sansohnin. Spinosin and feruloyl spinosin induced the prolongation of hexobarbital sleeping time in mice at doses of 50 and 100 mg/kg, respectively. We concluded that spinosin and feruloyl spinosin were the bioactive constituents of Sansohnin. It was assumed that spinosin and feruloyl spinosin circulated through the intestine and liver, therefore, these results will provide support for the sedative and hypnotic use of this crude drug in Oriental medicine. PMID- 9401727 TI - Effect of hachimi-jio-gan on immunoglobulin A producing cells in Peyer's patch by oral administration. AB - We investigated here the effect of Hachimi-jio-gan (HJ), a Japanese and Chinese herbal medicine, on immunoglobulin A (IgA) producing cells in Peyer's patch. The oral administration of HJ (0.1, 0.2, 1.0 g/kg for 2 d) enhanced the IgA producing cells almost 2-fold compared with the control group on days 4 and 5 after the administration. Furthermore, HJ augmented antigen-specific IgA (anti-SRBC IgA) production. These results suggested that HJ acted as a polyclonal B cell activator. To characterize its active ingredients, HJ was fractionated by ethanol precipitation. The crude polysaccharide fraction (EP fraction) showed the ability to augment IgA production, but low molecular weight fraction (ES fraction) did not. These results suggest that the crude polysaccharide fraction might be responsible for the augmentation of IgA production by HJ. PMID- 9401728 TI - Polysaccharide of Astragali radix enhances IgM antibody production in aged mice. AB - The effect of Astragali Radix (AR) on IgM antibody production in mice of various ages (10 weeks, 36 weeks and 60 weeks) was examined. The antibody production levels in the 36- and 60-week-old mice were significantly decreased to about 70 and 60% of that in the 10-week-old mice. The enhancement effect of a crude polysaccharide AR fraction on the antibody production was nil in the 10-week-old mice, but significant enhancement effects were observed in the 36- and 60-week old mice, compared to the age-matched control. Two polysaccharides active in the enhancement of the IgM antibody production in the aged mice were isolated from the high molecular weight fraction of AR by cetavlon precipitation, ion-exchange and gel permeation chromatography. The molecular masses of these polysaccharides were calculated by HPLC in salt solution. Only one major peak was observed for each, and their molecular masses were estimated to be 1.2 x 10(4) and 2.2 x 10(4). The major components of these polysaccharides were neutral carbohydrates (89.3 and 95.5%), followed by uronic acid and protein; glucose was the predominant sugar component. PMID- 9401729 TI - Osteotropic drug delivery system (ODDS) based on bisphosphonic prodrug. V. Biological disposition and targeting characteristics of osteotropic estradiol. AB - An osteotropic drug delivery system (ODDS) based on a bisphosphonic prodrug has been developed for 17 beta-estradiol (E2) to improve patient compliance in estrogen replacement therapy of postmenopausal osteoporosis. The biological disposition and the targeting efficiency of a bisphosphonic prodrug of E2, disodium [17 beta-(3'-hydroxy-1',3',5'-estratrienyloxy)carbonylpropyl carboxamidomethylene]bisphosphonate (E2-BP), was investigated in ovariectomized rats. After intravenous injection, E2-BP was rapidly taken up into the bone and subsequently cleared from the bone at a half-life of 13.5 d. The bone concentration of regenerated E2 was maintained throughout 28 d. In contrast, E2 injected intravenously showed extremely low bone distribution and rapid clearance from the bone, and E2 administered orally showed even lower bone distribution. Therapeutic availability (TA) and drug targeting index (DTI), which were calculated on the basis of the AUCs for E2 in the bone and plasma after injection of E2-BP and E2, were 64.6 and 451, respectively. These results suggest that ODDS has a potential to improve not only the apparent potency but also the therapeutic index of E2. As compared with the conventional estrogenic products, E2-BP should improve patient compliance with lower adverse effects and less frequent medication in long-term estrogen replacement therapy. PMID- 9401730 TI - Effect of aging on the intestinal transport of hydrophilic drugs in the rat small intestine. AB - The effect of aging on the intestinal transport of hydrophilic drugs (and probe compounds) was investigated in the rat small intestine. Passive transport was suggested to be unchanged with aging from 8 (young) to 54 (old) and further to 101 (very old) weeks old, as shown for D-xylose and urea in single-pass intestinal perfusion (under urethane anesthesia), where steady-state transport across the intestinal membrane into the blood stream was evaluated. The passive transports of cephradine, 5-fluorouracil (5-FU) and L-glucose were also unchanged, though they were compared only between the young and the old. Consistently, the passive uptake in the intestinal everted sacs, where the entry process into the membrane was evaluated for 5-FU, D-xylose, urea and polyethylene glycol (PEG) 900, was unchanged with aging from the young to the very old. The carrier-mediated transport of cephradine was also unchanged with aging from the young to the old in perfusion under anesthesia, though that of D-glucose was declined by about 50% with aging from the young to the old and thereafter remained constant in the very old. In perfusion in unanesthetized rats, age independency in passive transport (examined for cephradine, L-glucose and D xylose) and an age-dependent decline in D-glucose transport were also observed, suggesting that the findings under anesthesia are not qualitatively distorted. These results suggest that, although carrier-mediated transport may moderately decline with aging, the barrier function of the intestinal membrane to passive permeation of hydrophilic drugs (with molecular weight below 1000) may be unaffected by aging, supporting the suggestion from our previous in vivo studies that age-dependent increases in the orally absorbed fraction may be predicted for incompletely absorbed drugs because of delayed intestinal transit rather than increased intestinal transport (membrane permeability). PMID- 9401731 TI - Pharmacokinetics of pentazocine and its occupancy of opioid receptors in rat brain. AB - In order to assess quantitatively the pharmacodynamic process of pentazocine (PTZ), time courses of its plasma concentration and of the occupation of specific opioid receptors in the brain were investigated after intravenous (i.v.) administration of PTZ to rats. The plasma concentration of PTZ was determined by HPLC and the pharmacokinetic parameters were analyzed using nonlinear least squares analysis. Measurement of ex vivo receptor occupation was made by comparing the specific [3H]naloxone (opioid receptor antagonist) binding in vitro to the crude P2-synaptosomal fractions between vehicle-treated rats (control) and PTZ-treated rats. Following the i.v. administration of PTZ, the occupancy of specific opioid receptors decreased rapidly until 10 min, depending on the two pharmacological doses (2.5 and 10 mg/kg). The results strongly suggest the fast binding kinetics of PTZ in terms of its association with and dissociation from specific opioid receptor sites in the brain in addition to its fast rate of disappearance from the brain compartment. Furthermore, we demonstrated that the time profile of receptor occupancy correlated well (r = 0.8650) with that of the unbound concentration in plasma until 120 min after the i.v. administration of PTZ to rats. PMID- 9401732 TI - Metabolism of pravastatin sodium by 3 alpha-hydroxysteroid dehydrogenase. AB - When incubated with isolated rat hepatocytes, pravastatin sodium (PS) yielded a small amount of a metabolite in addition to two major metabolites that have already been reported. The previously uncharacterized metabolite was found to be formed by at first being enzymatically dehydrogenated to 6'-keto intermediate (R 104), followed by decomposition to give the aromatized metabolite (R-195), through spontaneous deesterification with accompanying aromatization. The PS 6'beta-hydroxydehydrogenase activity was localized in cytosolic fraction and required NADP, preferentially over NAD, as a cofactor. The formation of R-195 by rat liver cytosol was strongly inhibited by indomethacin, 3 alpha-hydroxysteroids (but not 3 beta-isomers) and 3-ketosteroids. The results and high substrate specificity of purified PS-6'beta-hydroxydehydrogenase toward 3 alpha hydroxysteroids suggested that the enzyme is identical to 3 alpha-hydroxysteroid dehydrogenase. PMID- 9401733 TI - Antibacterial activity of (+/-) 6-benzyl-1-(3-carboxypropyl) indane; a possible way to identify leading novel anti-H. pylori agents. AB - Magainin 2, isolated from the skin of the Xenopus laevis, is an antimicrobial peptide which reacts directly with the biological membrane to lyse various bacteria from negative and positive microorganisms. In a previous report, we showed that (+/-)1-(4-aminobutyl)-6-benzylindane (PM2), which mimicked the conformation of the side-chains of a complementary unit on the amino acid sequence of magainin 2 analogs, expressed the in vitro antibacterial activity not only against Helicobacter, pylori (ATCC43526, ATCC43579), but also against Escherichia coli (ATCC25922) and Staphylococcus aureus (ATCC25923). In addition, PM2 caused human blood red cells (RBCs) to lyse at the minimum inhibitory concentration (MIC) value. Based on the antibacterial activities of 9 phenylnonanoic acid (pC9c), we further synthesized (+/-)-6-benzyl-1-(3 carboxypropyl) indane (PM2c), which replaced a positive charge of PM2 with a negative one, and tested the biological activities. PM2c had the ability to inhibit the growth of H. pylori strains, but its activity to inhibit the growth of E. coli and S. aureus was not detected and weak, respectively. Moreover, PM2c showed non-hemolytic activity against RBCs at the MIC value. These results indicate the possibility that PM2c may be more useful than PM2 either alone or in combination with well-known therapeutic agents for the treatment of H. pylori infection. PMID- 9401734 TI - Determination of methemoglobin and carboxyhemoglobin in blood by rapid colorimetry. AB - A sensitive and rapid colorimetry for determination of methemoglobin (MetHb) in hemolysate of the test blood was devised by measurement of absorbance at 563 nm, the isosbestic point of spectra of cyanomethemoglobin and oxyhemoglobin, at pH 6.8. MetHb was determined as the difference in absorption caused by cyanide in the absence of potassium ferricyanide divided by the difference in absorption caused by cyanide in the presence of the ferricyanide. Carboxyhemoglobin (COHb) in the blood was also estimated from the absorbance values of the hemolysates with or without potassium ferricyanide after the addition of cyanide. The method requires only 3 microliters of test blood and 10 min for determinations of MetHb and COHb in blood. Results obtained by the method were in satisfactory agreement with theoretical results for mixture of MetHb, COHb, and oxyhemoglobin. The method was compared with two other methods using 55 forensic blood samples containing various amounts of MetHb and COHb, and proved to be suitable for practical samples. PMID- 9401735 TI - Characterization of two restriction endonucleases, SenPT14bI and SenPT16I, in standard phage-type strains of Salmonella enteritidis. AB - Two restriction endonucleases (ENases) were found by screening 38 standard phage strains of Salmonella (S.) Enteritidis. An isoschizomer of SacII ENase that recognizes the sequence 5'-CCGC/GG-3' was identified in S. Enteritidis PT14b, and an isoschizomer of XmaIII ENase (5'-C/GGCCG-3') was found in S. Enteritidis PT16. It is of special interest that the recognition specificities of all known ENases in Salmonella, including those of the S. Enteritidis ENases, are very similar to each other. PMID- 9401736 TI - Carbonyl reductase activity exhibited by pig testicular 20 beta-hydroxysteroid dehydrogenase. AB - The carbonyl reductase activity exhibited by pig testicular 20 beta hydroxysteroid dehydrogenase (20 beta-HSD) was examined using a recombinant enzyme. Kinetic parameters were obtained for 48 carbonyl group-containing substrates, including aromatic aldehydes, aromatic ketones, cycloketones, quinones, aliphatic aldehydes and aliphatic ketones. 20 beta-HSD showed a high affinity towards quinones, such as 9,10-phenanthrenequinone, alpha-naphthoquinone and menadione (Km values of 4, 2 and 5 microM, respectively), and the substrate utilization efficiency (Vmax/Km) of the enzyme against these quinones was very high. Cyclohexanone and 2-methylcyclohexanone were also reduced with a high Vmax/Km value, but not cyclopentanone or 2-methylcyclopentanone. Various aromatic aldehydes and ketones including benzaldehyde- and acetophenone-derivatives were reduced by 20 beta-HSD. Especially, 4-nitrobenzaldehyde and 4-nitroacetophenone were reduced with high Vmax/Km values in the related compounds. The enzyme also reduced the pyridine-derivatives, 2-, 3-, and 4-benzoylpyridine, with the Vmax/Km value for 2-benzoylpyridine being the highest. 20 beta-HSD reduced aliphatic aldehydes and aliphatic ketones, but was more effective on the former. The correlation between the structure of carbonyl compounds and their substrate Vmax/Km is discussed. PMID- 9401737 TI - Autoradiographic comparison of muscarinic M1 and M2 binding sites in the CNS of spontaneously hypertensive and normotensive rats. AB - Spontaneously hypertensive rats (SHR) respond with exaggerated pressor responses of central origin in response to pharmacologic stimulation of brain muscarinic receptors when compared with those to normotensive Wistar Kyoto (WKY) rats. At least part of the enhanced response to central muscarinic stimulation may be due to alterations in the expression of one or more of the five subtypes of muscarinic receptors. SHR are also known to exhibit regional alterations in the levels of mRNA encoding the M1, M2 and M4 receptors. In this study, we estimated the number of specific muscarinic receptor binding sites in 12-week-old SHR and WKY by measuring the binding of M1- and M2-selective ligands. Using standard autoradiographic techniques, coronal sections obtained from 12-week-old SHR and WKY were incubated with [3H]pirenzepine or [3H]AFDX 384 to label M1 and M2 receptors, respectively. Although both strains exhibited similar distribution patterns for both binding sites, sections derived from SHR expressed a significant increase in the number of [3H]pirenzepine binding sites compared to normotensive WKY in caudate putamen, CA3 region of the hippocampus, cingulate cortex, substantia nigra, posterior hypothalamic area and tuberomammillary nucleus. An increased number of [3H]AFDX 384 binding sites in SHR were observed in the olfactory tubercle, nucleus accumbens, basolateral amygdaloid nucleus, rostroventrolateral medulla and nucleus paragigantocellularis. Decreases in the number of [3H]AFDX 384 binding sites in SHR were also observed in the parietal cortex, medial geniculate, and lateral hypothalamic area. Statistically significant site-selective differences in binding densities between strains ranged from 4.0% to 35.5% of WKY means. These alterations in the expression of M1 and M2 binding sites in cardiovascular regions may contribute to the strain's hyper-responsiveness to cholinergic drugs and possibly to the appearance of other autonomic or behavioral phenotypes exhibited by SHR, including the hypertensive state itself. PMID- 9401738 TI - Mastoparan-induced apoptosis of cultured cerebellar granule neurons is initiated by calcium release from intracellular stores. AB - We have recently reported that mastoparan, a peptide toxin isolated from wasp venom, induces apoptosis in cultured cerebellar granule neurons that can be blocked by cholera toxin, an activator of Gs. Measurements of intracellular free calcium concentration ([Ca2+]i) reveal that mastoparan induces a dramatic elevation of [Ca2+]i that is frequently followed by enhanced leakage of fura-2 out of the neurons, suggesting that this rise in [Ca2+]i may be due to a more generalized change in membrane permeability. However, the mastoparan-induced initial elevation of [Ca2+]i is maintained in the absence of extracellular Ca2+, suggesting that the rise of [Ca2+]i is from intracellular stores. This conclusion is supported by the observation that depletion of [Ca2+]i stores by pretreatment with either caffeine or thapsigargin attenuates both the rise in [Ca2+]i and cell death induced by mastoparan. Phospholipase C (PLC) inhibitors, neomycin and U73122 block mastoparan-induced increases of [Ca2+]i and protect against neuronal death. Pretreatment with cholera toxin, but not pertussis toxin, reduced the mastoparan-induced rise in [Ca2+]i. Taken together, our data suggest that mastoparan initiates cell death in cerebellar granule neurons by inducing Ca2+ release from intracellular stores, probably via activation of PLC and IP3. A secondary or parallel process results in disruption of plasma membrane integrity and may be ultimately responsible for the death of these neurons by mastoparan. PMID- 9401739 TI - Sensory feedback contributes to early movement-evoked fields during voluntary finger movements in humans. AB - Neuromagnetic field changes accompanying voluntary movement in humans ('movement evoked fields' or MEFs) were recorded over the scalp using a whole-head MEG system during the performance of self-paced finger movements in order to determine the contribution of sensory feedback to the generation of these brain responses. It was found that cooling the subject's arm resulted in delays of 8 ms or more in the latency of the early movement-evoked field component (MEFI). These delays were attributed to increases in conduction times in the afferent pathways as confirmed by electrically evoked somatosensory responses and suggest a peripheral origin of the MEFI. In a second experiment, we demonstrated the effects of sensory input to the contralateral hand during a simple button pressing task in 4 subjects. The results indicated that responses over the hemisphere ipsilateral to the side of movement which resembled previously reported ipsilateral MEFs can be elicited by the spread of mechanical stimulation to opposite side of the body when a mechanical trigger is used. These experiments provide further evidence that early movement-evoked fields produced by unilateral finger movements are observed primarily over the contralateral somatosensory cortex and represent sensory feedback to the somatosensory cortex from the periphery. PMID- 9401740 TI - Axonal transport and distribution of cyclophilin A in chicken neurones. AB - In the course of pulse-label studies on the axonal transport of the small, basic, actin-binding proteins--actin depolymerizing factor, cofilin and profilin--in chicken motor neurones, we observed a heavily labelled protein of M(r) 18 kDa and pI 8.2 on fluorographs of two-dimensional polyacrylamide gels. On the basis of its M(r), pI and amino acid composition, we tentatively identified it by database searching as cyclophilin A and subsequently confirmed its identity by immunostaining. Like actin and its associated proteins, cyclophilin A was transported in slow component b of axonal transport, but unlike these proteins, cyclophilin A did not copurify with actin on DNase I. It was not found amongst labelled proteins transported by fast axonal transported by fast axonal transport. Immunostaining of chicken dorsal root ganglion cells revealed that it accumulated in neurites at points of branching, varicosities and growth cones. Our results raise the possibility that cyclophilin A is important in maintaining the native folding of actin and associated proteins during transit in axons and assembly in growth cones. PMID- 9401741 TI - Neurofibrillary lesions in experimental aluminum-induced encephalopathy and Alzheimer's disease share immunoreactivity for amyloid precursor protein, A beta, alpha 1-antichymotrypsin and ubiquitin-protein conjugates. AB - Neurofibrillary tangles of Alzheimer's disease contain predominantly tau protein and to a lesser degree amyloid precursor protein (APP), A beta protein, alpha 1 antichymotrypsin (ACT) and ubiquitin. Previously we have demonstrated the presence of phosphorylated tau and neurofilament proteins in neurofibrillary degeneration (NFD) induced by aluminum (Al) maltolate in rabbits [Savory et al., Brain Res. 669 (1995) 325-329; Savory et al., Brain Res. 707 (1996) 272-281]. Using the same animal system we have now detected APP, A beta, ACT and ubiquitin like immunoreactivities in NFD-bearing neurons, often colocalizing in the NFD. Diffuse cytoplasmic staining for APP, A beta and ubiquitin was also present in neurons without NFD from Al maltolate-treated rabbits. This study provides additional support for immunochemical similarities between Al-induced NFD in rabbits and the neurofibrillary tangles in human subjects with Alzheimer's disease. PMID- 9401742 TI - Methamphetamine-induced hyperthermia in mice: examination of dopamine depletion and heat-shock protein induction. AB - Methamphetamine (METH) is a common drug of abuse and a clinical anoretic which is known to cause neurotoxicity in rodents as evidenced by a depletion of dopamine (DA) and by decreased numbers of DA uptake sites in the striatum. It is also known to cause hyperthermia which is believed to induce the production of the 72 kDa heat-shock protein (HSP-72). In the present study, we evaluated whether METH induced the production of HSP-72 in both the mouse hippocampus and striatum and also attempted to correlate this induction with monoamine depletion. Adult male C57BL/6N mice received METH (20 mg/kg, i.p.) in an ambient temperature of 27 degrees C and body temperatures were monitored up to 240 min after treatment. Animals were sacrificed 12, 18, 24, 39, and 48 h after treatment. One striatum was examined for DA, DOPAC, and HVA levels using HPLC-EC and the contralateral striatum, along with the hippocampus, was prepared for immunoblotting. HPLC-EC analysis revealed a significant depletion of DA, DOPAC, and HVA at all time points. There was, however, a significant increase in DA at 48 vs. 39 h. A biphasic production of HSP-72, in both the hippocampus and striatum, was detected by immunoblot. HSP-72 production was strong at 12 h which corresponds to neuronal induction. However, at 18 h in the striatum and 24 h in the hippocampus, the induction appears to be reduced. A second phase of HSP-72 induction occurred at 39 h in both regions. In a second experiment, mice were dosed according to the same paradigm and were perfused at 18 h after treatment for immunohistochemical analysis. HSP-72 immunoreactivity was found in neurons of the CA1 and CA4 regions of the hippocampus; however, no detectable response was evident in the striatum. In conclusion, these data demonstrate that a single injection of METH can lead to hyperthermia which may then result in both the induction of HSP-72 and depletion of DA concentration. PMID- 9401743 TI - QX-314 inhibits ectopic nerve activity associated with neuropathic pain. AB - In animal models of neuropathic pain, transection or constrictive injury to peripheral nerves produces ectopic discharges originating at both injury sites and related dorsal root ganglia (DRG). In addition, hyperexcitability is observed in associated dorsal horn (DH) neurons of the spinal cord. As ectopic discharges are inhibited by agents that block voltage-sensitive Na+ channels, it has been postulated that accumulation of Na+ channels in the membrane at nerve injury sites may contribute to the development of ectopic nerve activity (ENA). The goal of the present study was to compare the sensitivity of ENA to lidocaine and QX 314, a positively charged lidocaine derivative, which is frequently assumed to be membrane impermeant. Experiments were performed on adult male Sprague-Dawley rats in which the common sciatic nerve had been transected 4-10 days earlier. Extracellular microelectrode recordings were made from DRG and DH neurons, and neuronal activity was measured in fine bundles of microfilaments teased from sciatic nerves in anesthetized and paralyzed rats. Comparative effects on heart rate (HR) and mean blood pressure (MBP) were also studied. To confirm that externally applied QX-314 is able to inhibit high frequency activity in sensory nerves, QX-314 was superfused over isolated rat vagus nerves during stimulation of compound action potentials in C-fibers (C-spikes). As expected, intravenously administered lidocaine inhibited ENA at all three sites. Lidocaine ED50 values (expressed as mg/kg, with 95% confidence limits) were: 10.2 (7.8-13.3), 1.4 (0.8 2.4) and 0.9 (0.4-2.0) for neuromas, DRG and DH neurons, respectively. QX-314 also induced dose-dependent inhibition of ENA at neuromas and DRG, but produced only a small inhibition of DH neuron ENA. QX-314 had the following ED50 values (mg/kg) for neuromas, DRG and DH neurons, respectively: 2.3 (2.0-2.8), 6.9 (4.7 26.5) and 85.7. QX-314-mediated inhibition of DRG ENA had a slow onset and was long-lasting, relative to lidocaine. Lidocaine or QX-314 also significantly reduced HR and MBP in the same dose range as that which reduced ENA in DRG or neuromas. In isolated rat vagus nerve recordings, QX-314 induced marked use dependent inhibition of C-spike amplitude, with IC50 values (microM) of 9000 (4600-18,000) and 350 (290-420) for low- (0.03 Hz) and high-frequency (30 Hz) C spikes, respectively. These data support the hypothesis that Na+ channel accumulation contributes to the generation of ectopic discharges in neuromas and DRG, and suggests that intravenous QX-314 can acutely block Na+ channels at these sites. PMID- 9401744 TI - Immunolocalization of the receptor tyrosine kinase EphA4 in the adult rat central nervous system. AB - EphA4 is a receptor tyrosine kinase of the Eph family previously designated Cek8 in chicken, Tyro1 in rat, and Sek1 in mouse, which is preferentially expressed in the embryonic and adult nervous system. We have mapped the distribution of EphA4 in the adult rat brain and spinal cord using a polyclonal antibody raised against a synthetic carboxy-terminal peptide. Immunoblotting experiments revealed that EphA4 is widely distributed in various regions of the adult rat brain. At the light microscopic level, intense immunoreactivity was apparent in the cerebral cortex, hippocampus, matrix compartment of the neostriatum, cholinergic neurons in the basal forebrain, cerebellar Purkinje cells, and substantia gelatinosa of the spinal cord. Among white matter tracts, EphA4 expression was detected in the corpus callosum, fornix, and posterior portion of the anterior commissure, but not in the lateral olfactory tract, mammillothalamic tract, or optic chiasm. Interestingly, expression in the optic chiasm is high at postnatal day 6, but decreases with the maturation of this structure. While in some regions of the neuropil neuronal cell bodies were prominently labeled, in others EphA4 immunoreactivity was detected in a punctate pattern. This punctuate staining did not coincide with synaptophysin localization. At the electron microscopic level, EphA4 immunoreactivity was observed in dendrites in the gray matter, particularly associated with dendritic spines, and in myelinated axons, but not their myelin sheaths in the white matter. The widespread distribution and diverse subcellular compartmentalization of EphA4 suggest that this receptor is important for the maintenance of multiple structures in the adult nervous system. PMID- 9401745 TI - A comparison of the effects of concentric versus eccentric exercise on force and position sense at the human elbow joint. AB - It is generally accepted that our sense of limb position and movement is provided, in part, by signals from muscle spindles, while the sense of muscle force derives from signals in tendon organs. Experiments are described here, using human subjects, in which the effects of eccentric and concentric exercise of elbow flexor muscles are compared on the sense of forearm position and the sense of tension in elbow flexors. Subjects were required to compress a preloaded spring with one arm, carrying out a concentric contraction in elbow flexors, then flexors of the other arm released the spring from compression and thereby carried out an eccentric contraction. The force of the spring was adjusted to be 20% maximum voluntary contraction (MVC), and each subject carried out a minimum of 120 contractions. Position sense was measured in blindfolded subjects by placing one forearm at a set angle and asking subjects to match it by positioning the other arm. Over 4 days postexercise, subjects placed the eccentrically exercised arms in a more extended position than the concentrically exercised arm suggesting that they thought the muscle was shorter than it actually was. In a force matching task, subjects systematically undershot the target 10% MVC with their eccentrically exercised arm. Since it is known that eccentric exercise is associated with damage to muscle fibres, it is postulated that this leads to a disturbance of muscle receptors, the muscle spindles and tendon organs. PMID- 9401746 TI - Neuromodulation of activity-dependent synaptic enhancement at crayfish neuromuscular junction. AB - Action potential-evoked transmitter release is enhanced for many seconds after moderate-frequency stimulation (e.g. 15 Hz for 30 s) at the excitor motorneuron synapse of the crayfish dactyl opener muscle. Beginning about 1.5 s after a train, activity-dependent synaptic enhancement (ADSE) is dominated by a process termed augmentation (G.D. Bittner, D.A. Baxter, Synaptic plasticity at crayfish neuromuscular junctions: facilitation and augmentation, Synapse 7 (1991) 235 243'[4]; K.L. Magleby, Short-term changes in synaptic efficacy, in: G.M. Edelman, L.E. Gall, C.W. Maxwell (Eds.), Synaptic Function, John Wiley and Sons, New York, 1987, pp. 21-56; K.L. Magleby; J.E. Zengel, Augmentation: a process that acts to increase transmitter release at the frog neuromuscular junction, J. Physiol. (Lond.) 257 (1976) 449-470) which decays approximately exponentially with a time constant of about 10 s at 16 degrees C, reflecting the removal of Ca2+ which accumulates during the train in presynaptic terminals (K.R. Delaney, D.W. Tank, R.S. Zucker, Serotonin-mediated enhancement of transmission at crayfish neuromuscular junction is independent of changes in calcium, J. Neurosci. 11 (1991) 2631-2643). Serotonin (5-HT, 1 microM) increases evoked and spontaneous transmitter release several-fold (D. Dixon, H.L. Atwood, Crayfish motor nerve terminal's response to serotonin examined by intracellular microelectrode, J. Neurobiol. 16 (1985) 409-424; J. Dudel, Modulation of quantal synaptic release by serotonin and forskolin in crayfish motor nerve terminals, in: Modulation of Synaptic Transmission and Plasticity in Nervous Systems, G. Hertting, H.-C. Spatz (Eds.), Springer-Verlag, Berlin, 1988; S. Glusman, E.A. Kravitz. The action of serotonin on excitatory nerve terminals in lobster nerve-muscle preparations, J. Physiol. (Lond.) 325 (1982) 223-241). We found that ADSE persists about 2-3 times longer after moderate-frequency presynaptic stimulation in the presence of 5-HT. This slowing of the decay of ADSE by 5-HT was not accompanied by significant changes in the initial amplitude of activity-dependent components of enhancement 1.5 s after the train. Measurements of presynaptic [Ca2+] indicated that the time course of Ca2+ removal from the presynaptic terminals after trains was not altered by 5-HT. Changes in presynaptic action potential shape, resting membrane potential or postsynaptic impedance after trains cannot account for slower recovery of ADSE. Axonal injection of EDTA slows the removal of residual Ca2+ and the decay of synaptic augmentation after trains of action potentials (K.R. Delaney, D.W. Tank, A quantitative measure of the dependence of short-term synaptic enhancement on presynaptic residual calcium, J. Neurosci. 14 (1994) 5885 5902), but has little or no effect on the 5-HT-induced persistence of ADSE. This also suggests that the time course of ADSE in the presence of 5-HT is not determined primarily by residual Ca2+ removal kinetics. The slowing of ADSE recovery after trains by 5-HT reverses with washing in 5-HT-free saline along with the 5-HT-mediated enhancement of release. PMID- 9401747 TI - Compartmentally specific effects of quinpirole on the striatal Fos expression induced by stimulation of D1-dopamine receptors in intact rats. AB - Injections of the full D1-agonist A-77636 (1.45 mg/kg) were found to induce clear Fos-like immunoreactivity (FLI) in the striatum of neurologically intact rats. Pretreatment with the D2-like agonist quinpirole (3 mg/kg) potentiated staining in the lateral striatum, but actually decreased the number of immunoreactive cells observed in the medial portion of the rostral striatum. Comparison with adjacent sections processed for the calcium binding protein calbindin, indicated that quinpirole pretreatment specifically suppressed staining in the matrix compartment of the striatum while tending to potentiate it in the striosomes, resulting in an extremely patchy pattern of labeling. These results suggest that exogenous stimulation of D2-receptors, although not essential for the induction of FLI, may play an important role in the compartmental patterning of neuronal activity within the striatum. PMID- 9401748 TI - Biphasic modulation in the trigeminal nociceptive neuronal responses by the intracerebroventricular prostaglandin E2 may be mediated through different EP receptors subtypes in rats. AB - To determine which prostaglandin E2 (PGE2) receptor subtypes are involved in the brain-derived PGE2-induced changes in nociception, we injected synthetic EP1, EP2 and EP3 receptor agonists (0.01 fmol to 10 nmol) into the lateral cerebroventricle (LCV) of urethane-anesthetized rats and observed the changes in the responses of the wide dynamic range (WDR) neurons in the trigeminal nucleus caudalis to noxious pinching of facial skin. The enhancement and suppression of the nociceptive responses of the WDR neurons were observed after the LCV injection of MB28767 (an EP3 receptor agonist) at a low dose range (1-100 fmol) and 17-phenyl-omega-trinor PGE2 (an EP1 receptor agonist) at high doses (1-10 nmol), respectively. Furthermore, the suppression of nociceptive neuronal responses after the LCV injection of PGE2 (1 nmol) was completely blocked by SC19220 (an EP1 receptor antagonist, 300 nmol). On the other hand, butaprost (an EP2 receptor agonist) at any doses tested (0.1 fmol to 1 nmol) had no effect on the nociceptive responses. The LCV injection of MB28767 (10 fmol) and 17-phenyl omega-trinor PGE2 (1 nmol), which respectively enhanced and suppressed the nociceptive neuronal responses, did not affect the responses of the low threshold mechanoreceptive neurons to innocuous tactile stimuli. These results provide electrophysiological evidence that brain-derived PGE2 induces mechanical hyperalgesia and hypoalgesia through EP3 and EP1 receptors, respectively, in the rat. PMID- 9401749 TI - The functional anatomy and evolution of hypoglossal afferents in the leopard frog, Rana pipiens. AB - Previously, we suggested that afferents are present in the hypoglossal nerve of the leopard frog, Rana pipiens. The basis for this was behavioral data obtained after transection of the hypoglossal nerve. These afferents coordinate the timing of tongue protraction with mouth opening during feeding. The goal of the present study was to define anatomically these hypoglossal afferents in Rana pipiens. Retrograde tracing was performed using horseradish peroxidase, fluorescent dextran amines and neurobiotin. Data show that the cell bodies of hypoglossal afferents are located in the dorsal root ganglion of the third spinal nerve and enter the brainstem through its dorsal root. The afferents ascend in the dorsomedial funiculus and move laterally after they pass the obex. They project in the granular layer of the cerebellum and the medial reticular formation. The cervical afferents that travel in this pathway are known to carry proprioceptive and cutaneous sensory information. We hypothesize that the presence of afferents in the hypoglossal nerve is a derived characteristic of anurans, which has resulted from the re-routing of afferent fibers from the third spinal nerve into the hypoglossal nerve. The appearance of hypoglossal afferents coincides with the morphological acquisition of a highly protrusible tongue. PMID- 9401750 TI - Two novel types of L-glutamate receptors with affinities for NMDA and L-cysteine in the olfactory organ of the Caribbean spiny lobster Panulirus argus. AB - A subset of olfactory receptor neurons of the Caribbean spiny lobster Panulirus argus possesses receptors for L-glutamate that can mediate both excitatory and inhibitory responses (P.C. Daniel, M.F. Burgess, C.D. Derby, Responses of olfactory receptor neurons in the spiny lobster to binary mixtures are predictable using a non-competitive model that incorporates excitatory and inhibitory transduction pathways, J. Comp. Physiol. A 178 (1992) 523-536). In this study, we have used biochemical and electrophysiological techniques to understand the role of these receptors in olfactory transduction, and to compare these olfactory glutamate receptors with peripheral and central L-glutamate receptors in other animals. Using a radioligand-binding assay with a membrane rich preparation from the dendrites of olfactory receptor neurons, we have identified two types of binding sites for L-glutamate. Both sites showed rapid, reversible, and saturable association with radiolabeled L-glutamate, and their Kd values (1 nM and 3 microM) are effective in physiological studies of glutamate sensitive olfactory neurons, suggesting these binding sites are receptors involved in olfactory transduction. Both sites were completely inhibited by high concentrations of NMDA and L-cysteine, and only partially inhibited by other L glutamate analogs and odorants. Electrophysiological recordings from L-glutamate best olfactory receptor neurons showed that NMDA and L-cysteine are both partial agonists and antagonists of glutamate receptors. Together, these results suggest the olfactory L-glutamate receptors of spiny lobsters are novel types of L glutamate receptors that are functionally important in mediating olfactory responses. PMID- 9401751 TI - Lisuride prevents learning and memory impairment and attenuates the increase in extracellular dopamine induced by transient global cerebral ischemia in rats. AB - In this experiment, we tested the efficacy of neuroprotection with lisuride, a dopamine agonist, using the 4-vessel occlusion rat model. Functional improvement was evaluated with two behavior tests exploring learning and memorization capacity in the rat, the Morris water maze and the 14-unit T-maze, 18 days after ischemia. Extracellular dopamine levels during ischemia were determined in search of a possible neuroprotection mechanism. Dopamine and its metabolites, DOPAC and HVA, as well as the serotonin metabolite, 5-HIAA, were assayed with HPLC-EC, in striatal extracellular fluid obtained by in vivo microdialysis in the awake rat. Lisuride was administered at a total dose of 10 ng by continuous intrastriatal infusion or at the dose of 0.5 mg/kg by i.p. infusion, 160 minutes before onset of ischemia for the neurochemical study and at the dose of 0.5 mg/kg via i.p. infusion, 1 hour before occlusion of the carotid arteries, for the behavior tests. Behavioral testing showed significantly better recovery in both sets of behavioral tests, with more pronounced positive results with the 14-unit T-maze, in comparison with the saline-treated animals. Microdialysis confirmed a significant attenuation of the ischemia-induced dopamine surge, whatever the mode of administration, compared with saline-treated animals. These results show that lisuride offers significant neuroprotection from the effect of experimental transient global forebrain cerebral ischemia in the rat; the mechanism would imply, at least in part, reduced levels of extracellular dopamine. PMID- 9401752 TI - Developmental cholinotoxicity of lead: loss of septal cholinergic neurons and long-term changes in cholinergic innervation of the hippocampus in perinatally lead-exposed rats. AB - The effects of perinatal lead exposure on choline acetyltransferase immunoreactive (ChAT-IR) cell counts in the medial septum and AChE-positive fiber counts in the hippocampus were examined in relation to changes in cholinergic markers in the septohippocampal pathway of the rat. Maternal exposure to 0.2% lead acetate in drinking water from gestational day 16 through weaning at post natal day 21 (P21) induced in the offspring a 30% reduction in septal ChAT activity and a 20% reduction in ChAT-IR cell profile counts in the medial septum/vertical diagonal band (MS/vDB). These changes were seen as early as P7, persisted through 2 months post-exposure (P81), and were followed by recovery of ChAT activity but not the ChAT-IR cell numbers, at 3 months post-exposure (P112). The loss of ChAT activity and ChAT-IR neurons in the septum was temporally associated with a reduction of ChAT activity (30%), hemicholinium-3 (HC-3) binding (40%), and AChE-positive fiber counts (13-15%) in the hippocampus. The hippocampal ChAT activity and AChE-positive fiber counts returned to control levels by P112 whereas HC-3 binding was restored to normal levels by P200. These results indicate that perinatal, low-level lead exposure induces loss of septohippocampal cholinergic projection neurons in neonate animals, resulting in a deficit in hippocampal cholinergic innervation that persists into young adulthood. The disruption of cholinergic septohippocampal system may be an important factor in lasting cognitive impairments associated with early Pb exposure. PMID- 9401753 TI - Morphological and functional characterization of an in vitro blood-brain barrier model. AB - Cell culture models have been extensively used for studies of blood-brain barrier (BBB) function. However, several in vitro models fail to reproduce some, if not most, of the physiological and morphological properties of in situ brain microvascular endothelial cells. We have recently developed a dynamic, tridimensional BBB model where endothelial cells exposed to intraluminal flow form a barrier to ions and proteins following prolonged co-culturing with glia. We have further characterized this cell culture model to determine whether these barrier properties were due to expression of a BBB phenotype. Endothelial cells of human, bovine or rodent origin were used. When co-cultured with glia, intraluminally grown endothelial cells developed features similar to in vivo endothelial cells, including tight junctional contacts at interdigitating processes and a high transendothelial resistance. This in vitro BBB was characterized by the expression of an abluminal, ouabain-sensitive Na/K pump, and thus favored passage of potassium ions towards the lumen while preventing K+ extravasation. Similarly, the in vitro BBB prevented the passage of blood-brain barrier-impermeant drugs (such as morphine, sucrose and mannitol) while allowing extraluminal accumulation of lipophylic substances such as theophylline. Finally, expression of stereo-selective transporters for Aspartate was revealed by tracer studies. We conclude that the in vitro dynamic BBB model may become an useful tool for the studies of BBB-function and for the testing of drug passage across the brain endothelial monolayer. PMID- 9401754 TI - Effects of noribogaine on the development of tolerance to antinociceptive action of morphine in mice. AB - The effects of noribogaine, a metabolite of ibogaine, on the development of tolerance to the antinociception action of morphine was determined in male Swiss Webster mice. Ibogaine is an alkaloid isolated from the bark of the African shrub, Tabernanthe iboga. Morphine tolerance in mice was developed by two different methods. Mice were rendered tolerant to morphine either by subcutaneous implantation of a pellet containing 25 mg morphine free base for 4 days or by injecting morphine (20 mg/kg, s.c.) twice a day for 4 days. Placebo pellet implanted mice or vehicle injected mice served as controls. To determine the effect of intraperitoneally administered noribogaine on tolerance development, the drug was injected in the appropriate dose twice a day. In pellet implanted mice, a dose of 20 mg/kg of noribogaine attenuated the tolerance to morphine whereas lower doses had no effect. Similarly, in mice given multiple injections of morphine, noribogaine attenuated tolerance development at 20 and 40 mg/kg doses. Previous studies from this laboratory had shown that ibogaine at 40 and 80 mg/kg doses inhibited tolerance to morphine. Because noribogaine could attenuate morphine tolerance at lower doses than ibogaine, it is concluded that the attenuating effect of ibogaine on morphine tolerance may be mediated by its conversion to noribogaine, a more active metabolite. PMID- 9401755 TI - Effect of aging on psychological stress-induced increases in noradrenaline release in the rat anterior hypothalamus: an in vivo microdialysis study. AB - Basal and psychological stress-induced noradrenaline (NA) release were studied by intracerebral microdialysis in the hypothalamus of rats aged 9 weeks or 12 months. Basal NA output was not significantly different between young (9-week old; 2.11 +/- 0.22 pg/20 min) and aged (12-month-old; 2.29 +/- 0.34 pg/20 min) rats. Psychological stress for 20 min significantly increased NA release in both groups (186% and 142% of baseline at 9 weeks old and 12 months old, respectively); however, the increase in aged rats was significantly lower than that in young rats (P < 0.01). This finding suggest that the noradrenergic neuronal response to psychological stress is attenuated in aged rats. PMID- 9401756 TI - Distribution of D-amino acid oxidase (DAO) activity in the medulla and thoracic spinal cord of the rat: implications for a role for D-serine in autonomic function. AB - The activity and regional distribution of D-amino acid oxidase (DAO), an enzyme that inactivates D-serine, were examined in the medulla and spinal cord of the rat by biochemical and histochemical procedures. DAO activity was noticeably low or absent in the nucleus of the solitary tract, ventrolateral medulla and intramediolateral cell column of the spinal cord. This may be indicative of a neuromodulatory role for endogenous D-serine (at the NMDA-glycine site) in in the central control of blood pressure. PMID- 9401757 TI - Potency and kinetics of nitric oxide-mediated vascular smooth muscle relaxation determined with flash photolysis of ruthenium nitrosyl chlorides. AB - Flash photolysis of thermally stable, photolabile 'caged' precursors permits rapid and precise changes of ligand concentration at their site of action. This approach was used to determine the concentration-dependence and time course of NO mediated relaxation of aortic smooth muscle, by use of two photolabile NO donors, trichloronitrosylruthenium (Ru(NO)Cl3) and dipotassium pentachloronitrosylruthenate (K2Ru(NO)Cl5). At concentrations up to 500 microM, both compounds were non-toxic before photolysis, and produced non-toxic by products on photolysis. Photolytic release of NO produced relaxations of intact and endothelium-denuded aortic rings precontracted with noradrenaline (0.1-0.5 microM), with an EC50 for NO-mediated relaxations of 10.5 nM and 13 nM, respectively. NO-mediated relaxations were reversibly blocked by 1 microM oxyhaemoglobin. The time course of NO-mediated relaxation comprised a delay of 3 7 s, followed by a sigmoidal decline in tension with peak rates that were strongly dependent on NO concentration. PMID- 9401758 TI - Dilatation by angiotensin II of the rat femoral arterial bed in vivo via pressure/flow-induced release of nitric oxide and prostaglandins. AB - 1. The haemodynamic effects of angiotensin II (AII) and, for comparison, arginine vasopressin (AVP) in the femoral and superior mesenteric artery of urethane anaesthetized rats were analysed with the ultrasonic transit time shift technique. 2. I.v. bolus injection of AII (0.1-3 nmol kg-1) and AVP (0.03-1 nmol kg-1) increased blood pressure which was accompanied by a decrease in blood flow through the superior mesenteric artery and an increase in femoral blood flow. The femoral hyperaemia was in part due to vasodilatation as indicated by a rise of femoral vascular conductance up to 200% relative to baseline. The femoral vasodilatation caused by AVP, but not AII, was followed by vasoconstriction. 3. Blockade of angiotensin AT1 receptors by telmisartan (0.2-20 mumol kg-1) prevented all haemodynamic responses to AII. 4. The femoral dilator responses to AII and AVP depended on the increase in vascular perfusion pressure since vasodilatation was reversed to vasoconstriction when blood pressure was maintained constant by means of a gravity reservoir. However, the AII-evoked femoral vasodilatation was not due to an autonomic or neuroendocrine reflex because it was not depressed by hexamethonium (75 mumol kg-1), prazosin (0.25 mumol kg-1) or propranolol (3 mumol kg-1). 5. The AII-induced femoral vasodilatation was suppressed by blockade of nitric oxide (NO) synthesis with NG nitro-L-arginine methyl ester (L-NAME, 40 mumol kg-1) and reversed to vasoconstriction when L-NAME was combined with indomethacin (30 mumol kg-1), but was left unaltered by antagonism of endothelin ETA/B receptors with bosentan (37 mumol kg-1). 6. These results demonstrate that the effect of AII to increase systemic blood pressure and the resulting rise of perfusion pressure in the femoral artery stimulates the formation of NO and prostaglandins and thereby dilates the femoral arterial bed. This local vasodilator mechanism is sufficient to mask the direct vasoconstrictor response to AII. PMID- 9401759 TI - Contrasting effects of carbachol, McN-A-343 and AHR-602 on Ca(2+)-mobilization and Ca(2+)-influx pathways in taenia caeci. AB - 1. We compared the binding profiles and contractile mechanisms of putative muscarinic M1 agonists McN-A-343 and AHR-602 with those of carbachol in smooth muscle of guinea-pig taenia caeci. 2. McN-A-343 and AHR-602, as well as carbachol, completely displaced the atropine-sensitive binding of [3H] quinuclidinyl benzilate to muscarinic receptors present in the membrane preparation. The potency order for the affinity of these agents for muscarinic receptors was carbachol > McN-A-343 >> AHR-602. 3. In the presence of 2.2 mM extracellular Ca2+, McN-A-343 and AHR-602 induced contraction corresponding to 79 and 85%, respectively, of the maximal contraction to 0.1 mM carbachol. Contractions induced by these agents were mediated via activation of the muscarinic receptor subtype that had a high affinity for 4-DAMP (M3 selective) but a low affinity for pirenzepine (M1 selective) and AF-DX 116 (M2 selective). These contractions were inhibited by an L-type Ca2+ channel blocker, verapamil. 4. In Ca(2+)-free solution containing 2 mM EGTA, carbachol elicited a transient contraction whereas no contraction was observed in response to McN-A-343 and AHR 602. Application of McN-A-343 or AHR-602 inhibited the carbachol-induced contraction in Ca(2+)-free solution, and this inhibition was surmounted by a higher concentration of carbachol. 5. The EC50 value for carbachol-induced contraction in the presence of extracellular Ca2+ was approximately 175 times lower than that in the absence of Ca2+. After treatment with propylbenzilylcholine mustard, carbachol induced contraction only in the presence of extracellular Ca2+. 6. The results suggest that in the taenia caeci there is a greater receptor reserve for muscarinic M3 receptor-mediated Ca2+ influx than for M3 mediated Ca2+ release. The compounds McN-A-343 and AHR-602 are agonists of the Ca2+ influx pathway, but do not appear to stimulate the Ca2+ release pathway. PMID- 9401760 TI - Subtypes of endothelin receptors that mediate venous effects of endothelin-1 in anaesthetized rats. AB - 1. The subtypes of endothelin receptors that mediate the effects of endothelin-1 (ET-1) on mean arterial pressure (MAP), heart rate (HR), mean circulatory filling pressure (MCFP), arterial resistance (RA), cardiac output (CO) and venous resistance (RV) were characterized in 9 groups of pentobarbitone-anaesthetized rats via the injection of ET-1 in the absence and presence of bosentan (Ro 47 0203, ETA- and ETB-receptor antagonist), PD 142893 (ETA- and ETB-receptor antagonist) or FR 139317 (ETA-receptor antagonist), as well as injection of the ETB-receptor agonist, IRL 1620. 2. Cumulative i.v. bolus injections of ET-1 or IRL 1620 (0.5, 1 and 2 nmol kg-1) dose-dependently increased MAP (ET: by 22, 34 and 44; IRL: 8, 17 and 28 mmHg), RA (ET: 62, 108 and 162; IRL: 51, 63 and 86% over baseline), RV (ET: 70, 132 and 179; IRL: 81, 89 and 98% over baseline) and MCFP (ET: 1.1, 1.8 and 1.9; IRL: 0.9, 1.0 and 1.2 mmHg) and reduced CO (ET: -18, 35 and -44; IRL: -24; -26; -25% below baseline). Equimolar doses of ET-1 and IRL 1620 caused similar initial transient depressor responses. Saline did not modify any haemodynamic variables in the time-control group. 3. Bosentan (10 mg kg-1, i.v.) inhibited ET-induced increases in MAP, RV, RA and MCFP and decrease in CO. PD 142893 (22 mg kg-1, i.v.) abolished ET-induced changes on MAP, RV, RA and CO, but did not alter effects on MCFP. Bosentan alone did not cause haemodynamic changes, but PD 142893 alone elevated MCFP (0.9 +/- 0.3 mmHg at 1 h after injection) and did not alter other variables. Both antagonists abolished the initial depressor effects of ET-1. 4. FR 139317 (1 mg kg-1, i.v.) partially inhibited the increases in MAP, RV, RA and MCFP and decreases in CO elicited by ET-1, but did not alter the transient depressor response of ET-1. 5. The results show that both ETA- and ETB-receptors mediate the arterial and venous constrictor effects of ET-1. Bosentan is more efficacious than PD 142893 in inhibiting the venous effects of ET-1. PMID- 9401761 TI - Morphological heterogeneity with normal expression but altered function of G proteins in porcine cultured regenerated coronary endothelial cells. AB - 1. Experiments were designed to investigate whether the pertussis toxin-dependent endothelial dysfunction following balloon injury is due to a reduced expression or an insufficient function of G-proteins. 2. Endothelium-dependent responses of porcine coronary arteries were examined in vitro by use of conventional organ chambers. Morphological analysis was performed by isolating and culturing the endothelial cells from these arteries. The expression of Gi-proteins in regenerated endothelial cells was measured by Western blots and immunolabelling. The function of G-proteins was assessed by measuring the GTPase activity of cultured endothelial cells. 3. Eight days following denudation, endothelial regrowth was confirmed by histological examination and by demonstrating the presence of endothelium-dependent relaxations to bradykinin and 5 hydroxytryptamine (5-HT). In primary culture, the regenerated endothelial cells displayed a 'cobblestone' pattern as seen with native endothelial cells. 4. Twenty eight days after denudation, the endothelium-dependent relaxations induced by 5-HT were impaired, but those to bradykinin were maintained. However, the latter were reduced when endothelium-dependent hyperpolarization was prevented. 5. Twenty eight days after denudation, multinucleated giant cells were present in the regenerated but not in the native cultured endothelial cell populations. These regenerated endothelial cells incorporated less tritiated thymidine than native endothelial cells. 6. The intensities of the bands on the immunoblot of the regenerated endothelial cells, when several antibodies against Gi alpha 1/alpha 2/alpha 3 were used, were the same as those obtained in native endothelial cells. The immunolabelling with the same antibodies was similar between the giant cells and the regenerated endothelial cells of normal size. The hydrolysis of GTP was lower in regenerated than in native endothelial cell membranes. 7. In conclusion, endothelium-dependent relaxations mediated by Gi proteins are impaired in balloon denuded coronary arteries. This dysfunction following regeneration cannot be explained by a reduced expression of Gi proteins but rather reflects an abnormal function of the G-proteins in the regenerated endothelium. PMID- 9401762 TI - Quantification and distribution of alpha 1-adrenoceptor subtype mRNAs in human vas deferens: comparison with those of epididymal and pelvic portions. AB - 1. This study was intended to quantify the amounts of the alpha 1-adrenoceptor subtype mRNAs in human vas deferens, and demonstrate the receptor subtype responsible for the vas contraction. 2. The RNase protection assay showed that the mean total amount of alpha 1a mRNA was 7.4 +/- 2.2 pg/5 micrograms of poly (A)+ RNA (97.0% of the total alpha 1 mRNA) in the epididymal portion (E-vas) and 4.9 +/- 0.8 pg/5 micrograms of poly (A)+ RNA (96.3% of the total) in the pelvic portion (P-vas). The E-vas showed a tendency to have a greater alpha 1a mRNA abundance than the P-vas (P = 0.11). The alpha 1b and alpha 1d mRNAs were absent or of extremely low abundance. 3. By an in situ hybridization, the alpha 1a and alpha 1d mRNAs were recognized in the smooth muscle cells of the E-vas and the P vas, and the distribution pattern the same in both tissue. The alpha 1b mRNA positive site was scarcely detectable in both vas portions. 4. In a functional study, l-phenylephrine produced concentration-dependent contraction in the E-vas (Emax = 2.24 +/- 0.70 g; pD2 = 5.32 +/- 0.09) and the P-vas (Emax = 2.46 +/- 0.46 g; pD2 = 5.07 +/- 0.12). KMD-3213, a novel alpha 1A-adrenoceptor-selective antagonist, caused parallel rightward shifts of the concentration-response curves for l-phenylephrine. Apparent pKB values were 9.90 +/- 0.16 for the E-vas and 9.71 +/- 0.17 for the P-vas. There was no significant difference in Emax, pD2 or pKB estimates between the two portions. 5. We have found that alpha 1a mRNA is predominant in the human vas deferens, and confirmed that contraction of this organ is mediated by the alpha 1A-adrenoceptor. PMID- 9401763 TI - Effect of thromboxane A2 antagonists on bronchial hyperresponsiveness induced immediately after interleukin-8 inhalation in guinea-pigs. AB - 1. Although repeated intranasal administration of interleukin-8 (IL-8) causes bronchial hyperresponsiveness (BHR) mediated via thromboxane A2 (TXA2) and airway neutrophil accumulation in guinea-pigs, the acute effect of inhaled IL-8 is unclear. We performed this study to clarify the acute effect of IL-8 on bronchial responsiveness and the role of TXA2. 2. The effects of inhaled IL-8 on bronchial responsiveness and of the TXA2 antagonists, S-1452 (0.01 and 0.1 mg kg-1) and ONO NT-126 (1.0 or 10 micrograms kg-1), on IL-8-induced BHR were examined by use of a modified Konzett-Rossler method in guinea-pigs. 3. Inhaled IL-8 at 100 ng ml-1, which failed to induce significant changes in Pao (pressure at the airway opening), enhanced an increase in Pao induced by subsequent inhalations of ascending doses (50-200 micrograms ml-1) of methacholine and histamine, suggesting the potentiating effect of IL-8 on bronchial responsiveness. No significant leukocyte infiltration was observed histologically sixteen minutes after the IL-8 inhalation. Both S-1452 and ONO-NT-126 reduced the IL-8-induced BHR. 4. In conclusion, IL-8 rapidly causes BHR via TXA2 release in guinea-pigs. PMID- 9401764 TI - Fidelity in functional coupling of the rat P2Y1 receptor to phospholipase C. AB - 1. The rat homologue of the P2Y1 receptor has been heterologously expressed in 1321N1 human astrocytoma cells and in C6 rat glioma cells. 2. As has been shown previously for the turkey and human P2Y1 receptors, the rat P2Y1 receptor expressed in either cell type responded to 2MeSATP with increases in inositol phosphate accumulation that were competitively blocked by the antagonist PPADS. Neither of the wild type cell lines exhibited inositol phosphate responses to P2Y1 receptor agonists. 3. Expression of the rat P2Y1 receptor did not confer a capacity of 2MeSATP to inhibit adenylyl cyclase activity in 1321N1 cells. Moreover, the inhibition of adenylyl cyclase mediated by an endogenous P2Y receptor of C6 glioma cells was not enhanced by expression of the rat P2Y1 receptor. The P2Y receptor-mediated inhibition of adenylyl cyclase in C6 glioma cells expressing both the endogenous P2Y receptor and the rat P2Y1 receptor remained unaffected by PPADS. 4. Since the P2Y receptor responsible for inhibition of adenylyl cyclase in C6 glioma cells does not share the pharmacological or functional properties of the P2Y1 receptor, even when both receptors originate from the same species and are simultaneously expressed in the same cell line, it is concluded that the P2Y1 receptor is distinct from an endogenous P2Y receptor in C6 cells that couples to inhibition of adenylyl cyclase. PMID- 9401765 TI - Hexamethonium- and methyllycaconitine-induced changes in acetylcholine release from rat motor nerve terminals. AB - 1. The neuronal nicotinic receptor antagonists hexamethonium and methyllycaconitine (MLA) have been used to study the putative prejunctional nicotinic ACh receptors (AChRs) mediating a negative-feedback control of ACh release from motor nerve terminals in voltage-clamped rat phrenic nerve/ hemidiaphragm preparations. 2. Hexamethonium (200 microM), but not MLA (0.4-2.0 microM), decreased the time constant of decay of both endplate currents (e.p.cs) and miniature endplate currents (m.e.p.cs), indicating endplate ion channel block with hexamethonium. However, driving function analysis and reconvolution of e.p.cs and m.e.p.cs indicated that this ion channel block did not compromise the analysis of e.p.c. quantal content. 3. At low frequencies of stimulation (0.5-2 Hz), hexamethonium (200 microM) and MLA (2.0 microM) increased e.p.c. quantal content by 30-40%. At high frequencies (50-150 Hz) neither compound affected e.p.c. quantal content. All effects on quantal content were paralleled by changes in the size of the pool of quanta available for release. 4. The low frequency augmentation of e.p.c. quantal content by hexamethonium was absent when extracellular [Ca2+] was lowered from 2.0 to 0.5 mM. 5. At the concentrations studied, MLA and hexamethonium produced a small (10-20%) decrease in the peak amplitude of m.e.p.cs. 6. Neither apamin (100 nM) nor charybdotoxin (80 nM) had effects on spontaneous or nerve evoked current amplitudes at any frequency of stimulation. Thus the ability of nicotinic antagonists to augment e.p.c. quantal content is not due to inhibition of Ca(2+)-activated K(+)-channels. 7. We suggest that hexamethonium and MLA increase evoked ACh release by blocking prejunctional nicotinic AChRs. These receptors exert a negative feedback control over evoked ACh release and are probably of the alpha-bungarotoxin-insensitive neuronal type. PMID- 9401766 TI - Ethanol inhibition of N-methyl-D-aspartate-activated current in mouse hippocampal neurones: whole-cell patch-clamp analysis. AB - 1. The action of ethanol on N-methyl-D-aspartate (NMDA)-activated ion current was studied in mouse hippocampal neurones in culture using whole-cell patch-clamp recording. 2. Ethanol inhibited NMDA-activated current in a voltage-independent manner, and did not alter the reversal potential of NMDA-activated current. 3. Concentration-response analysis of NMDA- and glycine-activated current revealed that ethanol decreased the maximal response to both agonists without affecting their EC50 values. 4. The polyamine spermine (1 microM) increased amplitude of NMDA-activated current but did not alter the percentage inhibition of ethanol. 5. Compared to an extracellular pH of 7.0, pH 6.0 decreased and pH 8.0 increased the amplitude of NMDA-activated current, but these changes in pH did not significantly alter the percentage inhibition by ethanol. 6. The sulphydryl reducing agent dithiothreitol (2 mM) increased the amplitude of NMDA-activated current, but did not affect the percentage inhibition by ethanol. 7. Mg2+ (10, 100, 500 microM), (5, 20 microM) or ketamine (2, 10 microM) decreased the amplitude of NMDA-activated current, but did not affect the percentage inhibition by ethanol. 8. The observations are consistent with ethanol inhibiting the function of NMDA receptors by a non-competitive mechanism that does not involve several modulatory sites on the NMDA receptor-ionophore complex. PMID- 9401767 TI - Nitrergic modulation of cholinergic responses in the opossum lower oesophageal sphincter. AB - 1. Electrical field stimulation (EFS) of the superfused lower oesophageal sphincter from opossum (Monodelphis domestica) elicited biphasic responses. The first phase (relaxation) was strictly dependent on the duration of the EFS. The second phase (contraction) started following termination of the EFS (< or = 15 Hz). EFS at frequencies above 15 Hz led only to contraction, which started immediately upon initiation of the stimulation. 2. In the presence of NG-nitro-L arginine (L-NOARG; 0.1-300 microM), the relaxation phase was abolished and the contractile response started with the initiation of EFS (at all frequencies) and was greater in magnitude. The contractile response to EFS was completely blocked with scopolamine (10 microM). 3. Exogenous acetylcholine (1-100 microM) elicited concentration-dependent contractions of the sphincter in the presence of botulinum toxin. These contractions were abolished when EFS was applied during administration of acetylcholine. This inhibitory effect of EFS was completely reversed when the tissue was treated with L-NOARG (100 microM). 4. These results suggest that the cholinergic response in the opossum lower oesophageal sphincter is under nitrergic control. PMID- 9401768 TI - Effect of pentoxifylline on the degradation of procollagen type I produced by human hepatic stellate cells in response to transforming growth factor-beta 1. AB - 1. Pentoxifylline (PTF) may act as a potential antifibrogenic agent by inhibiting cell proliferation and/or collagen deposition in cell type(s) responsible for the accumulation of extracellular matrix. The aim of the present study was to investigate at which level PTF may affect synthesis and degradation of type I collagen in human hepatic stellate cells (HSCs), a key source of connective tissue in fibrotic liver. 2. Procollagen type I synthesis and release were evaluated in cells maintained in serum free/insulin free medium for 48 h and then stimulated with transforming growth factor-beta 1 (TGF-beta 1) for different time periods in the presence or absence of PTF. TGF-beta 1 caused an upregulation of procollagen I mRNA levels with a peak increase after 3-6 h of stimulation. This effect was followed by an increase in both the cell associated and the extracellular levels of the corresponding protein, with a peak effect at 9-12 h after the addition of TGF-beta 1. Co-incubation with PTF slightly but consistently reduced basal as well as stimulated procollagen I mRNA levels, with negligible effects on the cell-associated expression of the corresponding protein. Conversely, PTF dose-dependently reduced procollagen type I levels detected in supernatants from unstimulated and stimulated cells. 3. Pulse-chase experiments employing L-[3H]-proline revealed that PTF was able to induce significantly the degradation of procollagen, mainly in the extracellular compartment. We next analysed the effect of PTF on the major pathway involved in type I collagen degradation. PTF did not affect the expression of metalloproteinase 1 (MMP-1) mRNA both in basal and stimulated conditions, whereas it markedly reduced the expression of tissue inhibitor of metalloproteinase 1 (TIMP-1) mRNA. Accordingly incubation with PTF increased the levels of 'activated MMP-1' in cell supernatants in both basal and stimulated conditions. 4. These results suggest that the antifibrogenic action of PTF on human HSCs is mainly mediated by extracellular collagen degradation rather than by a reduction of collagen synthesis. PMID- 9401769 TI - Effects of dexamethasone and phorbol ester on P2 receptor-coupled Ca2+ signalling and lipocortin 1 presentation in U937 cells. AB - 1. Cell surface bound lipocortin 1 (LC1) is a putative mediator of the antiproliferative and anti-inflammatory effects of glucocorticoids. This study assessed the hypothesis that the glucocorticoid, dexamethasone-phosphate (dex-p), might exert the above effects via an LC1-mediated downregulation of receptor coupled Ca2+ signalling, using P2-receptor mediated intracellular Ca2+ accumulation in U937 cells as an appropriate model. 2. Addition of ATP (1-100 microM) to cells resulted in a transient increase in cytosolic Ca2+ ([Ca2+]i). Prior treatment of cells with dex-p (3-24 h) increased the magnitude of this Ca2+ transient at high, but not low concentrations of ATP, and increased thapsigargin (Tg)-induced Ca2+ influx, indicating that store-operated Ca2+ influx was potentiated in these cells. For cells treated with dex-p for 24 h, cell surface levels of LC1 were significantly reduced by 63%. 3. Differentiation of cells with 1 nM phorbol ester (PMA) for 24 h resulted in a 2.4 fold increase in the cell surface level of LC1 and inhibition of the ATP-induced Ca2+ response. However, the Tg-induced Ca2+ response was unaffected by long-term PMA treatment, and incubating cells with LC1 did not alter Tg-induced Ca2+ mobilization and influx, or the ATP-mediated Ca2+ response. 4. Data from this study suggest that: (1) dex p does not inhibit P2-receptor-coupled Ca2+ signalling in this cell line, (2) the observed modulation of the ATP-induced increase in [Ca2+]i by dex-p and PMA, and store-operated Ca2+ influx by dex-p, is not linked to an increase in the cell surface level of LC1, and (3) differentiation of U937 cells with PMA downregulates the ATP-induced Ca2+ response, but does not affect the thapsigargin sensitive Ca2+ pool or store-operated Ca2+ influx of these cells. PMID- 9401770 TI - Tranilast inhibits the proliferation, chemotaxis and tube formation of human microvascular endothelial cells in vitro and angiogenesis in vivo. AB - 1. First developed as an antiallergic drug, tranilast inhibits chemical mediator release from mast cells. In the present study, we examine the effects of tranilast on angiogenesis in vitro and in vivo and discuss the application of tranilast for angiogenic diseases. 2. Tranilast inhibited significantly the proliferation (IC50: 136 microM, 95% confidence limits: 134-137 microM) and vascular endothelium growth factor (VEGF)-induced chemotaxis (IC50: 135 microM, 95% confidence limits: 124-147 microM) of human dermal microvascular endothelial cells (HDMECs) at concentrations greater than 25 micrograms ml-1. No toxicity to HDMECs measuring by LDH release and no inhibitory effects on metalloproteinase (MMP)-2 and MMP-9 activity were observed even at 100 micrograms ml-1 (306 microM). 3. Tube formation of HDMECs cultured on the matrigel as an in vitro angiogenesis model was inhibited by tranilast in a concentration-dependent manner. The IC50 value and 95% confidence limits were 175 microM and 151-204 microM, respectively. 4. In vivo angiogenesis was induced in mice by the subcutaneous injection of matrigel containing 30 ng ml-1 VEGF and 64 micrograms ml-1 heparin. Tranilast was administered orally twice a day for 3 days. Tranilast dose-dependently suppressed angiogenesis in the matrigel and a significant change was observed at a dose of 300 mg kg-1. 5. These results indicate that tranilast is an angiogenesis inhibitor which may be beneficial for the improvement of angiogenic diseases such as proliferative diabetic retinopathy, age-related macular degeneration, tumour invasion and rheumatoid arthritis. PMID- 9401771 TI - Beta-adrenoceptor-mediated inhibition of alpha 1-adrenoceptor-mediated and field stimulation-induced contractile responses in the prostate of the guinea pig. AB - 1. The prostate of the guinea pig responds to electrical field-stimulation (2 s trains, 0.1 ms pulses at 3-60 Hz, supramaximal voltage) with contractile responses. At 18 Hz these responses were inhibited (82 +/- 2%) by the L-type Ca2+ channel blocker, nifedipine (10 microM) and (by 100%) by the neurotoxin, tetrodotoxin (500 nM). The alpha 1A-selective adrenoceptor antagonist, 5 methylurapidil, inhibited responses to field stimulation in the absence and presence of nifedipine (10 microM) with -log molar (p) IC50 (+/- s.e. mean) values of 7.95 +/- 0.14 and 7.01 +/- 0.07, respectively. 2. The non-selective beta-adrenoceptor agonist, isoprenaline, reduced (56 +/- 8%) field stimulation induced contractile responses (pEC50 6.91 +/- 0.11). The non-selective beta adrenoceptor antagonist propranolol (50 nM) and the beta 1-adrenoceptor selective antagonist, atenolol (3 microM), but not the beta 2-adrenoceptor antagonist ICI 118,551 ((+/-)-1 -[2,3-(dihydro-7-methyl-1H-inden-4-yl)oxyl]-3-[1 methylethyl)amino ]-2-butanol HCl; 100 nM) antagonized this effect (apparent pKB values 8.44 +/- 0.22 and 6.92 +/- 0.21, respectively) indicating an effect mediated through beta 1-like adrenoceptors. In the presence of nifedipine (10 microM) isoprenaline (up to 10 microM) did not inhibit the remaining response to field-stimulation. 3. Phenylephrine elicited contractile responses (pEC50 4.47 +/ 0.30) from preparations of guinea pig prostate which were reduced (63 +/- 25%) by nifedipine (10 microM). This response was antagonized by 5-methylurapidil (100 nM, apparent pKB 8.24 +/- 0.33), but was not affected by preincubation chloroethylclonidine (50 microM, 30 min). Responses to phenylephrine (30 microM) were inhibited (by up to 52 +/- 5%) by isoprenaline (pIC50 6.40 +/- 0.35, the beta 2-adrenoceptor selective agonist, salbutamol was weakly effective). Propranolol (300 nM), ICI 118,551 (100 nM) and atenolol (3 microM) shifted isoprenaline concentration-response curves to the right (apparent pKB +/- s.e. values 7.68 +/- 1.10; 8.00 +/- 0.72 and 6.62 +/- 0.95, respectively). In the presence of nifedipine (10 microM) responses to phenylephrine (30 microM,) were inhibited (by up to 51 +/- 4%) by isoprenaline (pIC50 6.88 +/- 0.17): propranolol (300 nM) and ICI 118,551 (100 nM), but not atenolol (3 microM) antagonized this effect (apparent pKB values 8.85 +/- 1.53 and 8.35 +/- 1.18, respectively). Thus beta 1-like and beta 2-like adrenoceptors may be involved in the isoprenaline stimulated inhibition of phenylephrine concentration-response curves. 4. Phenylephrine stimulated [3H]-inositol phosphate accumulation (pEC50 4.47 +/- 0.83), an effect insensitive to chloroethylclonidine pre-treatment (50 microM, 30 min) and to nifedipine (10 microM), but inhibited by 5-methylurapidil (apparent pKD 7.90 +/- 0.22). Isoprenaline (up to 1 microM) did not affect the phenylephrine-stimulated maximal increase in [3H]-inositol phosphates but did increase [3H]-cyclic adenosine monophosphate ([3H]-cAMP) accumulation (pEC50 6.77 +/- 0.66); propranolol (30 nM) and ICI 118,551 (110 nM), but not atenolol (up to 3 microM), antagonized this effect. These responses may therefore be mediated through beta 2-like adrenoceptors. 5. These results show that the alpha 1 adrenoceptor mediated and field stimulation-induced contractions of the guinea pig prostate are partly dependent upon intracellular and extracellular sources of Ca2+. We conclude that both beta 1- and beta 2-like adrenoceptors inhibit responses to phenylephrine in the prostate of the guinea pig. The beta 1-like adrenoceptor-mediated inhibition of these responses is evident upon the field stimulation-induced and nifedipine-sensitive component of the response to phenylephrine and may not involve the activation of adenylyl cyclase. The beta 2 like adrenoceptor may inhibit both nifedipine sensitive and insensitive components of the response to phenylephrine, possibly through the activation of adenylyl cyclase, but not through the i PMID- 9401772 TI - Calcium mobilization in Jurkat cells via A2b adenosine receptors. AB - 1. A functional study of cell surface A2b adenosine receptors was performed on the T cell leukaemia line, Jurkat. 2. A2b receptors were coupled both to the adenylate cyclase system and to intracellular calcium channels. In fact, the agonist of A2b receptors, 5'-N-ethylcarboxamidoadenosine (NECA), led to a transient accumulation of intracellular calcium by an inositol phosphate independent mechanism. 3. The NECA-induced accumulation of cGMP was not responsible for the calcium mobilization via A2b receptors. 4. The calcium response elicited by activation of A2b receptors was independent of that evoked by activation of the T cell receptor. 5. These findings not only delineate a novel transduction mechanism for adenosine but also support a specific role for adenosine in modulating signals elicited via the T cell receptor. PMID- 9401773 TI - Effect of peroxynitrite on plasma extravasation, microvascular blood flow and nociception in the rat. AB - 1. Peroxynitrite (ONOO-) is a cytotoxic species, formed by the reaction between nitric oxide and superoxide free radicals, that may be involved in inflammation. In this study we have investigated the effect of peroxynitrite on plasma extravasation and microvascular blood flow in the dorsal skin and on nociceptive responses in the hind paw of the rat. 2. Male Wistar rats were anaesthetized and their dorsal skin shaved. Plasma extravasation was measured by the extravascular accumulation of 125I-labelled albumin over 0-45 min and 0-240 min. Blood flow was measured by laser-Doppler flowmetry over 0-240 min. Studies in the hind paw were carried out in the conscious rat. Hind paw weight changes were determined by volume displacement and nociception by a mechanical hyperalgesia technique. 3. Intradermal (i.d.) peroxynitrite (100-200 nmol site-1) produced a significant (P < 0.01) dose-dependent increase in plasma extravasation in dorsal skin over 0-45 min which was not increased over 45-240 min. Plasma extravasation was significantly (P < 0.001) decreased in rats pretreated with the anti-inflammatory steroid dexamethasone (1 mg kg-1, i.v.; -180 min), but not modulated by treatment with the hydrogen peroxide deactivator catalase (2200 u site-1), or the superoxide scavenger superoxide dismutase (500 u site-1), effective doses of the tachykinin NK1 antagonist SR140333 (1 nmol site-1), the cyclo-oxygenase inhibitor indomethacin (358 mumol site-1), or combined pretreatment with mepyramine (histamine H1-receptor antagonist; 2.8 nmol site-1) and methysergide (5-HT antagonist; 1.9 nmol site-1). 4. Microvascular blood flow was significantly (P < 0.05) increased 30 and 120 min after i.d. peroxynitrite (100 nmol site-1) in dorsal skin and remained raised until the end of the recording period (240 min). The increase in blood flow was unaffected by dexamethasone (1 mg kg-1, i.v.; -180 min) or indomethacin (10 mg kg-1, s.c.; -30 min). 5. Hind paw volume was significantly (P < 0.001) increased 30 min after intraplantar peroxynitrite (87.5 and 175 nmol paw-1) and remained raised for the duration of the experiment (360 min). By comparison, nociception was not altered by intraplantar peroxynitrite. 6. These data indicate that peroxynitrite can cause an increase in both plasma extravasation and blood flow, suggesting that peroxynitrite could be of biological relevance to microvascular responses. These findings may be of importance in the pathology of inflammatory diseases in which peroxynitrite formation occurs. PMID- 9401774 TI - Interaction of the renin-angiotensin system, bradykinin and sympathetic nerves with cholinergic transmission in the rat isolated trachea. AB - 1. The present study was undertaken to investigate the interaction of the renin angiotensin system (RAS), bradykinin and the sympathetic nervous system with cholinergic transmission in the rat airways. Experiments were performed on epithelium-intact and epithelium-denuded preparations of rat isolated trachea which had been incubated with [3H]-choline to incorporate [3H]-acetylcholine into the cholinergic transmitter stores. Tracheal preparations were subjected to electrical field stimulation (trains of 1 ms pulses, 5 Hz, 15 V) and the stimulation-induced (S-I) efflux taken as an index of transmitter acetylcholine release. 2. In both epithelium-intact and epithelium-denuded tracheal preparations, the alpha 2-adrenoceptor agonist UK14304 (0.1 and 1 microM) inhibited the S-I efflux, in a concentration-dependent manner. The inhibition of S-I efflux produced by UK14304 (1 microM) was antagonized by the selective alpha 2-adrenoceptor antagonist idazoxan (0.3 microM). Idazoxan (0.3 microM) alone had no effect on the S-I efflux. 3. Angiotensin II (0.1 and 1 microM) was without effect on the S-I efflux in either epithelium-intact or epithelium-denuded tracheal preparations. When angiotensin-converting enzyme was inhibited by perindoprilat (10 microM), angiotensin II (1 microM) was also without effect on the S-I efflux. Similarly, in the presence of idazoxan (0.3 microM), to block prejunctional alpha 2-adrenoceptors, angiotensin II (0.1 and 1 microM) did not alter the S-I efflux. When added alone, perindoprilat (10 microM) did not alter the S-I efflux. 4. In epithelium-denuded preparations, bradykinin (0.01-1 microM) inhibited the S-I efflux. In epithelium-intact preparations, there was also a tendency for bradykinin (0.1 and 1 microM) to inhibit the S-I efflux but this was not statistically significant. However, when angiotensin-converting enzyme and neutral endopeptidase were inhibited by perindoprilat (10 microM) and phosphoramidon (1 microM), respectively, bradykinin (1 microM) significantly inhibited the S-I efflux in epithelium-intact preparations as well as in epithelium-denuded preparations. The inhibition of the S-I efflux produced by bradykinin, in the combined presence of perindoprilat (10 microM) and phosphoramidon (1 microM), was unaffected by the additional presence of the cyclo oxygenase inhibitor indomethacin (10 microM) and/or the nitric oxide synthase inhibitor NG-nitro-L-arginine (100 microM), in either epithelium-intact or epithelium-denuded preparations. 5. In conclusion, the findings of the present study suggest that airway parasympathetic nerves are endowed with alpha 2 adrenoceptors which subserve inhibition of transmitter acetylcholine release. Under the present conditions, however, transmitter acetylcholine release is not subject to transneuronal modulation by noradrenaline released from adjacent sympathetic nerves in the airways. Moreover, angiotensin II and perindoprilat do not appear to modulate acetylcholine release from parasympathetic nerves of the airways. In contrast, bradykinin inhibits acetylcholine release from airway parasympathetic nerves but this action of bradykinin is limited by the activity of epithelial angiotensin-converting enzyme and/or neutral endopeptidase. The inhibitory action of bradykinin on cholinergic transmission in the airways does not appear to involve the liberation of prostaglandins or nitric oxide. PMID- 9401775 TI - The influence of 5-HT2 and 5-HT4 receptor antagonists to modify drug induced disinhibitory effects in the mouse light/dark test. AB - 1. The ability of 5-HT2 and 5-HT4 receptor antagonists to modify the disinhibitory profile of diazepam and other agents was investigated in male BKW mice in the light/dark test box. 2. The 5-HT2A/2B/2C receptor antagonists ritanserin, MDL11939 and RP62203 and also methysergide, which failed to modify mouse behaviour when administered alone, caused dose-related enhancements (4 to 8 fold) in the potency of diazepam to disinhibit behavioural responding to the aversive situation of the test box. 3. Ritanserin was shown to enhance the disinhibitory potency of other benzodiazepines, chlordiazepoxide (4 fold), temazepam (10 fold) and lorazepam (10 fold), the 5-HT1A receptor ligands, 8-OH DPAT (25 fold), buspirone (100 fold) and lesopitron (500 fold), the 5-HT3 receptor antagonists, ondansetron (100 fold) R(+)-zacopride (100 fold) and S(-) zacopride (greater than a 1000 fold), the substituted benzamides, sulpiride (10 fold) and tiapride (5 to 10 fold) and the cholecystokinin (CCK)A receptor antagonist, devazepide (100 fold). It also reduced the onset of action of disinhibition following treatment with the 5-HT synthesis inhibitor parachlorophenylalanine. Ritanserin failed to enhance the disinhibitory effects of the CCKB receptor antagonist CI-988, the angiotensin AT1 receptor antagonist losarten or the angiotensin converting enzyme inhibitor ceranapril. 4. The 5-HT4 receptor antagonists SDZ205-557, GR113808 and SB204070 caused dose-related reductions in the disinhibitory effect of diazepam, returning values to those shown in vehicle treated controls. The antagonists failed to modify mouse behaviour when administered alone. 5. GR113808 was also shown to cause a dose related antagonism of the disinhibitory effects of chlordiazepoxide, lorazepam, 8 OH-DPAT, buspirone, lesopitron, ondansetron, R(+)-zacopride, sulpiride, tiapride, devazepide, CI-988, losarten, ceranapril and parachlorophenylalanine. 6. It was concluded that in BKW mice (a) the failure of 5-HT2 and 5-HT4 receptor antagonists when administered alone to modify behaviour in the light/dark test indicates an absence of an endogenous 5-HT tone at the 5-HT2 and 5-HT4 receptors and (b) the enhancement by the 5-HT2 receptor antagonists and attenuation by the 5-HT4 receptor antagonists of drug-induced disinhibition indicates a plurality of 5-HT receptor involvement in the mediation of drug-induced disinhibitory profiles in the mouse. PMID- 9401776 TI - Binding and effects of KATP channel openers in the vascular smooth muscle cell line, A10. AB - 1. The ATP-sensitive K+ channel (KATP channel) in A10 cells, a cell line derived from rat thoracic aorta, was characterized by binding studies with the tritiated KATP channel opener, [3H]-P1075, and by electrophysiological techniques. 2. Saturation binding experiments gave a KD value of 9.2 +/- 5.2 nM and a binding capacity (BMax) of 140 +/- 40 fmol mg-1 protein for [3H]-P1075 binding to A10 cells; from the BMax value a density of binding sites of 5-10 per microns2 plasmalemma was estimated. 3. KATP channel modulators such as the openers P1075, pinacidil, levcromakalim and minoxidil sulphate and the blocker glibenclamide inhibited [3H]-P1075 binding. The extent of inhibition at saturation depended on the compound, levcromakalim inhibiting specific [3H]-P1075 binding by 85%, minoxidil sulphate and glibenclamide by 70%. The inhibition constants were similar to those determined in strips of rat aorta. 4. Resting membrane potential, recorded with microelectrodes, was -51 +/- 1 mV. P1075 and levcromakalim produced a concentration-dependent hyperpolarization by up to -25 mV with EC50 values of 170 +/- 40 nM and 870 +/- 190 nM, respectively. The hyperpolarization induced by levcromakalim (3 microM) was completely reversed by glibenclamide with an IC50 value of 86 +/- 17 nM. 5. Voltage clamp experiments were performed in the whole cell configuration under a physiological K+ gradient. Levcromakalim (10 microM) induced a current which reversed around -80 mV; the current-voltage relationship showed considerable outward rectification. Glibenclamide (3 microM) abolished the effect of levcromakalim. 6. Analysis of the noise of the levcromakalim (10 microM)-induced current at -40 and -20 mV yielded estimates of the channel density, the single channel conductance and the probability of the channel to be open of 0.14 micron-2, 8.8 pS and 0.39, respectively. 7. The experiments showed that A10 cells are endowed with functional KATP channels which resemble those in vascular tissue; hence, these cells provide an easily accessible source of channels for biochemical and pharmacological studies. The density of binding sites for [3H]-P1075 was estimated to be one order of magnitude higher than the density of functional KATP channels; assuming a plasmalemmal localization of the binding sites this suggests a large receptor reserve for the openers in A10 cells. PMID- 9401777 TI - Tumour necrosis factor-alpha expression and cell recruitment in Sephadex particle induced lung inflammation: effects of dexamethasone and cyclosporin A. AB - 1. Tumour necrosis factor-alpha (TNF-alpha) is a cytokine with diverse properties consistent with a possible role in inflammatory disease. We investigated whether TNF-alpha is induced during the progression of lung inflammation elicited by a particulate non-antigenic stimulus, and whether pharmacological control of TNF alpha expression influences recruitment of specific inflammatory cell types. 2. A single intravenous injection of Sephadex particles into rats led to extensive granulomatous inflammation in lung alveolar and bronchial tissue that peaked in intensity after 24-72 h. Mononuclear cells were the principal component of granulomas, but neutrophils and eosinophils were also abundant. Numbers of mononuclear cells, neutrophils and eosinophils recovered by bronchoalveolar lavage (BAL) peaked at 72 h, 48 h and 72 h, respectively. 3. Messenger RNA encoding TNF-alpha was induced in lung epithelial cells, lung granulomas and BAL cells 6 h after Sephadex administration and remained elevated for 72 h before declining to baseline by 7 days. In BAL cell populations TNF-alpha protein was localized to mononuclear cells at all times points pre- and post-Sephadex administration. 4. Treatment of rats with dexamethasone significantly reduced the Sephadex-induced recruitment of mononuclear cells, neutrophils and eosinophils into the bronchoalveolar cavity, and significantly reduced TNF-alpha mRNA expression by BAL cells. 5. Treatment of rats with cyclosporin A was without effect on Sephadex-induced elevations of mononuclear cell numbers and expression of TNF-alpha, but did reduce significantly recruitment of neutrophils and eosinophils to BAL cell populations. 6. These results show that a sequential asthma-like recruitment of neutrophils, eosinophils and mononuclear cells into lung tissue can be induced by single exposure to a non-antigenic stimulus. Pharmacological and histological studies reveal that mononuclear cell mobilization relates closely to induced TNF-alpha expression, whereas mobilization of neutrophils and eosinophils appears secondary to expression of the cytokine. PMID- 9401778 TI - Investigation of the role of nitric oxide and cyclic GMP in both the activation and inhibition of human neutrophils. AB - 1. The aim of this study was to establish the role of nitric oxide (NO) and cyclic GMP in chemotaxis and superoxide anion generation (SAG) by human neutrophils, by use of selective inhibitors of NO and cyclic GMP pathways. In addition, inhibition of neutrophil chemotaxis by NO releasing compounds and increases in neutrophil nitrate/nitrite and cyclic GMP levels were examined. The ultimate aim of this work was to resolve the paradox that NO both activates and inhibits human neutrophils. 2. A role for NO as a mediator of N-formyl-methionyl leucyl-phenylalanine (fMLP)-induced chemotaxis was supported by the finding that the NO synthase (NOS) inhibitor L-NMMA (500 microM) inhibited chemotaxis; EC50 for fMLP 28.76 +/- 5.62 and 41.13 +/- 4.77 pmol/10(6) cells with and without L NMMA, respectively. Similarly the NO scavenger carboxy-PTIO (100 microM) inhibited chemotaxis; EC50 for fMLP 19.71 +/- 4.23 and 31.68 +/- 8.50 pmol/10(6) cells with and without carboxy-PTIO, respectively. 3. A role for cyclic GMP as a mediator of chemotaxis was supported by the finding that the guanylyl cyclase inhibitor LY 83583 (100 microM) completely inhibited chemotaxis and suppressed the maximal response; EC50 for fMLP 32.53 +/- 11.18 and 85.21 +/- 15.14 pmol/10(6) cells with and without LY 83583, respectively. The same pattern of inhibition was observed with the G-kinase inhibitor KT 5823 (10 microM); EC50 for fMLP 32.16 +/- 11.35 and > 135 pmol/10(6) cells with and without KT 5823, respectively. 4. The phosphatase inhibitor, 2,3-diphosphoglyceric acid (DPG) (100 microM) which inhibits phospholipase D, attenuated fMLP-induced chemotaxis; EC50 for fMLP 19.15 +/- 4.36 and 61.52 +/- 16.2 pmol/10(6) cells with and without DPG, respectively. 5. Although the NOS inhibitors L-NMMA and L-canavanine (500 microM) failed to inhibit fMLP-induced SAG, carboxy-PTIO caused significant inhibition (EC50 for fMLP 36.15 +/- 7.43 and 86.31 +/- 14.06 nM and reduced the maximal response from 22.14 +/- 1.5 to 9.8 +/- 1.6 nmol O2-/10(6) cells/10 min with and without carboxy-PTIO, respectively). This suggests NO is a mediator of fMLP induced SAG. 6. A role for cyclic GMP as a mediator of SAG was supported by the effects of G-kinase inhibitors KT 5823 (10 microM) and Rp-8-pCPT-cGMPS (100 microM) which inhibited SAG giving EC50 for fMLP of 36.26 +/- 8.77 and 200.01 +/- 43.26 nM with and without KT 5823, and 28.35 +/- 10.8 and 49.25 +/- 16.79 nM with and without Rp-8-pCTP-cGMPS. 7. The phosphatase inhibitor DPG (500 microM) inhibited SAG; EC50 for fMLP 33.93 +/- 4.23 and 61.12 +/- 14.43 nM with and without DPG, respectively. 8. The NO releasing compounds inhibited fMLP-induced chemotaxis with a rank order of potency of GEA 3162 (IC50 = 14.72 +/- 1.6 microM) > GEA 5024 (IC50 = 18.44 +/- 0.43 microM) > SIN-1 (IC50 > 1000 microM). This order of potency correlated with their ability to increase cyclic GMP levels rather than the release of NO, where SIN-1 was most effective (SIN-1 (EC50 = 37.62 +/- 0.9 microM) > GEA 3162 (EC50 = 39.7 +/- 0.53 microM) > GEA 5024 (EC50 = 89.86 +/- 1.62 microM)). 9. In conclusion, chemotaxis and SAG induced by fMLP can be attenuated by inhibitors of phospholipase D, NO and cyclic GMP, suggesting a role for these agents in neutrophil activation. However, the increases in cyclic GMP and NO induced by fMLP, which are associated with neutrophil activation, are very small. In contrast much larger increases in NO and cyclic GMP, as observed with NO releasing compounds, inhibit chemotaxis. PMID- 9401779 TI - Inhibition by levetiracetam of a non-GABAA receptor-associated epileptiform effect of bicuculline in rat hippocampus. AB - 1. Extracellular recording of field potentials, evoked by commissural stimulation in hippocampal area CA3 of anaesthetized rats, was performed in order to study the mode of action of the novel antiepileptic drug levetiracetam (ucb LO59). 2. The amplitude of orthodromic field population spike (PS2) markedly increased and repetitive population spikes appeared when the recording micropipette contained either bicuculline methiodide (BMI), or the specific GABAA antagonist gabazine (SR-95531). 3. BMI-induced increases in PS2 were reduced in a dose-dependent manner by 1 to 320 mumol kg-1 levetiracetam i.v., with a U-shape dose-response relationship. However, levetiracetam did not reduce the increases in PS2 produced by gabazine. 4. Clonazepam (1 mg kg-1, i.p.), carbamazepine (20 mg kg-1, i.p.) and valproate (200 mg kg-1, i.v.) were ineffective in preventing BMI-induced increases in PS2, while the calcium channel antagonist flunarizine, 50 mumol kg 1, i.p., reduced PS2 increments caused by BMI. The L-type calcium channel blocker nifedipine, 100 mumol kg-1, i.p., was without effect. Similar to levetiracetam, flunarizine did not reduce the increases in PS2 induced by gabazine. 5. These data suggest that the increased excitability of CA3 neurones, caused by BMI administered in situ, involves calcium-dependent processes not associated with blockade of GABAA receptors. The inhibition by levetiracetam of this calcium dependent effect of BMI might contribute to the antiepileptic effects of the drug. PMID- 9401780 TI - Electrophysiological characterisation of tachykinin receptors in the rat nucleus of the solitary tract and dorsal motor nucleus of the vagus in vitro. AB - 1. Recent studies have shown antagonists at the NK1 subtype of receptor for tachykinins are antiemetics and suggested that this may result from blockade of tachykinin-mediated synaptic transmission at a central site in the emetic reflex. 2. We have used intracellular recording in vitro to study the pharmacology of tachykinins in the nucleus of the solitary tract (NST) and dorsal motor nucleus of the vagus (DMNV). 3. Neurones in the NST were depolarized by substance P (SP), the presumed endogenous ligand for the NK1 receptor and these effects were mimicked by the NK1 agonists, SP-O-methylester (SPOMe), GR73632 and septide; however, SP was nearly an order of magnitude less potent than the latter two agonists. 4. In the DMNV, SP and NK1 receptor agonists evoked similar depolarising responses but SP appeared to be more potent than in the NST and was closer in potency to the other agonists. 5. NK1-receptor antagonists blocked responses to septide and GR73632 in the NST but had little effect on responses to SP and SPOMe. In contrast, in the DMNV the NK1-receptor antagonists blocked responses to septide and GR73632 but also reduced responses to SP and SPOMe. 6. Neurokinin A (NKA) was almost equipotent with septide and GR73632 in depolarizing both NST and DMNV neurones but these effects were not mimicked by a specific NK2 receptor agonist. Responses to NKA were unaffected by an NK2-receptor antagonist; however, the depolarizing effects of NKA were blocked by NK1-receptor antagonists. 7. Neurones in both DMNV and NST were unaffected by the endogenous NK3-receptor ligand, neurokinin B and by a specific agonist for this site, senktide. 8. The results with NK1 receptor agonists and antagonists suggest that the septide-sensitive NK1 site is involved in the excitation of both NST and DMNV neurones. The 'classical' NK1 receptor may play more of a role in the DMNV and a third unknown site may be responsible for the depolarizing response to SP in the NST. The effects of NKA are best interpreted as an action at the septide sensitive NK1 site. This raises the possibility that anti-emetic action of the NK1 antagonists may be due to blockade of NKA transmission at the septide sensitive site. PMID- 9401781 TI - Cardiac inotropes inhibit the oedema caused by nifedipine in rabbit skin. AB - 1. We have shown previously that exposing the rat or rabbit microcirculation to nifedipine increases the permeability of the post-capillary venule, the segment of microcirculation that is known to control inflammatory oedema. 2. In the present study modulation by the inotropes isoprenaline, dopexamine and dobutamine of nifedipine-induced oedema was examined in the rabbit skin microcirculation by measuring the localised leakage of 125I-radiolabelled albumin after the i.d. injection of agents. 3. Coinjection of isoprenaline (10(-11) moles per site), dopexamine (10(-10) moles per site) or dobutamine (10(-10) moles per site) suppressed significantly (P < 0.05) the oedema response to nifedipine (10(-7.2) moles per site) in the rabbit dorsal skin microcirculation. 4. To confirm the oedema suppresser effect of the inotropes, dopexamine or dobutamine were also coinjected with histamine 10(-8) + PGE2 10(-10) moles per site, or bradykinin 10( 10) + PGE2 10(-10) moles per site. Both inotropes at 10(-10) moles per site reduced significantly (P < 0.05) the leakage of albumin caused by bradykinin + PGE2 and histamine + PGE2. 5. When measured by laser Doppler, basal local skin blood flow increased at 30 min by 57 +/- 14% with nifedipine 10(-7.2) moles per site and 15 +/- 11% with isoprenaline 10(-11) moles per site. Isoprenaline did not suppress the blood flow response to nifedipine, the response to coinjection being 68 +/- 11%. 6. Oedema caused by nifedipine can be suppressed by low concentrations of beta-adrenergic agonists that do not suppress the blood flow response to nifedipine. This suggests that cardiac inotropes can influence non inflammatory changes in microvascular permeability. PMID- 9401782 TI - Activation of the cAMP transduction cascade contributes to the mechanical hyperalgesia and allodynia induced by intradermal injection of capsaicin. AB - 1. The spinal role of the cAMP transduction cascade in nociceptive processing was investigated in awake behaving rats (male, Sprague-Dawley) by activating or inhibiting this pathway spinally. Microdialysis fibres were implanted into the dorsal horn to infuse drugs directly to the spinal cord. 2. Animals, without peripheral tissue injury, were tested for responses to repeated applications (10 trials) of von Frey filaments and threshold to mechanical stimulation before and after infusion of 8-bromo-cAMP. In this group of animals treated spinally with 8 br-cAMP (1-10 mM) a dose-dependent hyperalgesia and allodynia were produced. This was manifested as an increased number of responses to 10 trials of von Frey filaments (10, 50, 150, 250 mN) and a decrease in mechanical threshold. 3. A second series of experiments studied the manipulation of the cAMP pathway spinally in a model of tissue injury induced by intradermal injection of capsaicin. Animals were either pre- or post-treated spinally with the adenylate cyclase inhibitor, tetrahydrofuryl adenine (THFA) or the protein kinase A inhibitor, myrosilated protein kinase (14-22) amide (PKI). Injection of capsaicin resulted in an increased number of responses to repeated applications of von Frey filaments and a decrease in threshold to mechanical stimuli outside the site of injection, secondary mechanical hyperalgesia and allodynia. 4. Pre-treatment with either THFA (1 mM) or PKI (5 mM) had no effect on the capsaicin-evoked secondary hyperalgesia and allodynia. 5. In contrast, post-treatment spinally with THFA (0.01-1 mM) or PKI (0.05-50 mM) dose-dependently reduced the mechanical hyperalgesia and allodynia produced by capsaicin injection. Furthermore, the mechanical hyperalgesia and allodynia blocked by the adenylate cyclase inhibitor, THFA (1 mM), was reversed by infusion of 8-bromo-cAMP (0.01-10 mM) in a dose dependent manner. 6. Thus, this study demonstrates that activation of the cAMP transduction cascade at the spinal cord level results in mechanical hyperalgesia and allodynia and that the secondary mechanical hyperalgesia and allodynia following intradermal injection of capsaicin is mediated by this same transduction cascade. PMID- 9401783 TI - Evidence for a 5-HT3 receptor involvement in the facilitation of peristalsis on mucosal application of 5-HT in the guinea pig isolated ileum. AB - 1. The 5-HT receptor involved in the effect of mucosal application of 5-HT to facilitate peristalsis was investigated in the isolated guinea pig ileum. 2. An application of 5-HT (3-100 microM) to the mucosal surface (by inclusion of 5-HT in the Krebs-Henseleit solution passing through the lumen of the ileum) caused a concentration related facilitation of peristalsis characterized by a reduction in the peristaltic threshold. 3. Peristalsis was not modified by methiothepine (0.1 microM), ritanserin (0.1 microM), ondansetron (5 microM), granisetron (1 microM) or SB 204070 (0.1 microM) administered alone to the mucosal surface. 4. The concentration-response curve to mucosally applied 5-HT was not altered by the mucosally applied 5-HT1/2 receptor antagonist methiothepine (0.1 microM), the 5 HT2 receptor antagonist ritanserin (0.1 microM) or the 5-HT4 receptor antagonist SB 204070 (0.1 microM). However, the mucosally applied 5-HT3 receptor antagonists ondansetron (5 microM) and granisetron (1 microM) shifted the response curves to mucosally applied 5-HT to the right in a parallel and surmountable manner. The pD2 values in the absence and presence of ondansetron were 5.42 +/- 0.07 and 4.12 +/- 0.10, respectively, (n = 6) and that of granisetron were 5.45 +/- 0.12 and 4.50 +/- 0.10 respectively, (n = 5). 5. Serosally applied ondansetron (5 microM) or granisetron (1 microM) had no effect on the concentration-response curve to mucosally applied 5-HT. However, the serosally applied ondansetron and granisetron antagonised the facilitatory effect of serosally applied 5-HT (10 microM) when administered in the presence of serosally applied SB 204070 (0.1 microM). 6. It is concluded that the facilitatory effect of mucosally applied 5 HT to reduce the peristaltic threshold in the guinea pig ileum is mediated via a 5-HT3 receptor located on the mucosal and not the serosal side of the ileum. PMID- 9401784 TI - Intravascular and interstitial degradation of bradykinin in isolated perfused rat heart. AB - 1. Bradykinin (BK) has been shown to exert cardioprotective effects which are potentiated by inhibitors of angiotensin I-converting enzyme (ACE). In order to clarify the significance of ACE within the whole spectrum of myocardial kininases we investigated BK degradation in the isolated rat heart. 2. Tritiated BK (3H-BK) or unlabelled BK was either repeatedly perfused through the heart, or applied as an intracoronary bolus allowing determination of its elution kinetics. BK metabolites were analysed by HPLC. Kininases were identified by ramiprilat, phosphoramidon, diprotin A and 2-mercaptoethanol or apstatin as specific inhibitors of ACE, neutral endopeptidase 24.11 (NEP), dipeptidylaminopeptidase IV and aminopeptidase P (APP), respectively. 3. In sequential perfusion passages, 3H BK concentrations in the perfusate decreased by 39% during each passage. Ramiprilat reduced the rate of 3H-BK breakdown by 54% and nearly abolished [1-5] BK generation. The ramiprilat-resistant kininase activity was for the most part inhibited by the selective APP inhibitor apstatin (IC50 0.9 microM). BK cleavage by APP yielded the intermediate product [2-9]-BK, which was rapidly metabolized to [4-9]-BK by dipeptidylaminopeptidase IV. 4. After bolus injection of 3H-BK, 10% of the applied radioactivity were protractedly eluted, indicating the distribution of this fraction into the myocardial interstitium. In samples of such interstitial perfusate fractions, 3H-BK was extensively (by 92%) degraded, essentially by ACE and APP. The ramiprilat- and mercaptoethanol-resistant fraction of interstitial kininase activity amounted to 14%, about half of which could be attributed to NEP. Only the product of NEP, [1-7]-BK, was continuously generated during the presence of 3H-BK in the interstitium. 5. ACE and APP are located at the endothelium and represent the predominant kininases of rat myocardium. Both enzymes form a metabolic barrier for the extravasated fraction of BK. Thus, only interstitial, but not intravascular concentrations of BK are increased by kininase inhibitors to the extent that a significant potentiation of BK effects could be explained. NEP contributes less than 5% to the total kininase activity, but is the only enzyme which is exclusively present in the interstitial space. PMID- 9401785 TI - Fc epsilon RI-mediated chloride uptake by rat mast cells: modulation by chloride transport inhibitors in relation to histamine secretion. AB - 1. We have examined the role of extracellular chloride in the mast cell secretion process. The immunologically-directed ligand, antibody to IgE (anti-IgE) required extracellular chloride ions for optimum secretion from rat peritoneal mast cells. In contrast, replacement of extracellular chloride did not alter the mast cell secretory response to compound 48/80, calcium ionophore A23187 or substance P. 2. Anti-IgE-stimulation of mast cells evoked a significant uptake of chloride ions compared to non-stimulated cells. The magnitude of chloride uptake correlated with the magnitude of stimulated histamine secretion. 3. Compound 48/80, substance P and A23187 did not alter the rate of chloride ion uptake, although these agents caused significant histamine secretion. 4. The Na+/K+/2Cl- cotransport inhibitor, furosemide, reduced the rate of anti-IgE-stimulated chloride uptake at a relatively high concentration (700 microM). However, the more potent Na+/K+/2Cl- cotransport inhibitors, bumetanide (100 microM) and piretanide (100 microM) had no effect on the stimulated chloride uptake. 5. Furosemide inhibited anti-IgE-induced histamine secretion, bumetanide potentiated the response and piretanide had no effect. This suggests that their respective action on histamine secretion are unrelated to inhibition of the Na+/K+/2Cl- carrier. 6. The chloride channel blocker, 5-nitro-2-((3-phenylpropyl)-amino) benzoic acid (NPPB), reduced both anti-IgE-stimulated chloride uptake and the corresponding histamine secretion in a dose-dependent manner. The magnitude of the inhibitory action of the drug on these two cellular processes was comparable, implying that chloride channel activity is related to the mechanism of histamine secretion. 7. It is concluded that chloride uptake has a role in the control of Fc epsilon RI-mediated histamine secretion from rodent mast cells. PMID- 9401786 TI - Modulation of nicotinic ACh-, GABAA- and 5-HT3-receptor functions by external H 7, a protein kinase inhibitor, in rat sensory neurones. AB - 1. The effects of external H-7, a potent protein kinase inhibitor, on the responses mediated by gamma-aminobutyric acid A type (GABAA)-, nicotinic acetylcholine (nicotinic ACh)-, ionotropic 5-hydroxytryptamine (5-HT3)-, adenosine 5'-triphosphate (ATP)-, N-methyl-D-aspartate (NMDA)- and kainate (KA) receptors were studied in freshly dissociated rat dorsal root ganglion neurone by use of whole cell patch-clamp technique. 2. External H-7 (1-1000 microM) produced a reversible, dose-dependent inhibition of whole cell currents activated by GABA, ACh and 5-HT. 3. Whole-cell currents evoked by ATP, 2-methylthio-ATP, NMDA and KA were insensitive to external H-7. 4. External H-7 shifted the dose-response curve of GABA-activated currents downward without changing the EC50 significantly (from 15.0 +/- 4.0 microM to 18.0 +/- 5.0 microM). The maximum response to GABA was depressed by 34.0 +/- 5.3%. This inhibitory action of H-7 was voltage independent. 5. Intracellular application of H-7 (20 microM), cyclic AMP (1 mM) and BAPTA (10 mM) could not reverse the H-7 inhibition of GABA-activated currents. 6. The results suggest that external H-7 selectively and allosterically modulates the functions of GABAA-, nicotine ACh- and 5-HT3 receptors via a common conserved site in the external domain of these receptors. PMID- 9401787 TI - Species differences in brain adenosine A1 receptor pharmacology revealed by use of xanthine and pyrazolopyridine based antagonists. AB - 1. The pharmacological profile of adenosine A1 receptors in human, guinea-pig, rat and mouse brain membranes was characterized in a radioligand binding assay by use of the receptor selective antagonist, [3H]-8-cyclopentyl-1,3-dipropylxanthine ([3H]-DPCPX). 2. The affinity of [3H]-DPCPX binding sites in rat cortical and hippocampal membranes was similar. Binding site affinity was higher in rat cortical membranes than in membranes prepared from guinea-pig cortex and hippocampus, mouse cortex and human cortex. pKD values (M) were 9.55, 9.44, 8.85, 8.94, 8.67, 9.39 and 8.67, respectively. The binding site density (Bmax) was lower in rat cortical membranes than in guinea-pig or human cortical membranes. 3. The rank order of potency of seven adenosine receptor agonists was identical in each species. With the exception of 5'-N-ethylcarboxamidoadenosine (NECA), agonist affinity was 3.5-26.2 fold higher in rat cortical membranes than in human and guinea-pig brain membranes; affinity in rat and mouse brain membranes was similar. While NECA exhibited 9.3 fold higher affinity in rat compared to human cortical membranes, affinity in other species was comparable. The stable GTP analogue, Gpp(NH)p (100 microM) reduced 2-chloro-N6-cyclopentyladenosine (CCPA) affinity 7-13.9 fold, whereas the affinity of DPCPX was unaffected. 4. The affinity of six xanthine-based adenosine receptor antagonists was 2.2-15.9 fold higher in rat cortical membranes compared with human or guinea-pig membranes. The rank order of potency was species-independent. In contrast, three pyrazolopyridine derivatives, (R)-1-[(E)-3-(2-phenylpyrazolo[1,5-a]pyridin-3-yl) acryloyl]-2-piperidine ethanol (FK453), (R)-1-[(E)-3-(2-phenylpyrazolo[1,5 a]pyridin-3-yl) acryloyl]-piperidin-2-yl acetic acid (FK352) and 6-oxo-3-(2 phenylpyrazolo[1,5-a]pyridin-3-yl)-1(6H)-pyridazinebutyric acid (FK838) exhibited similar affinity in human, guinea-pig, rat and mouse brain membranes. pKi values (M) for [3H]-DPCPX binding sites in human cortical membranes were 9.31, 7.52 and 7.92, respectively. 5. Drug affinity for adenosine A2A receptors was determined in a [3H]-2-[4-(2-carboxyethyl)phenethylamino]-5'-N-ethylcarboxamido ade nosine ([3H]-CGS 21680) binding assay in rat striatal membranes. The pyrazolopyridine derivatives, FK453, FK838 and FK352 exhibited pKi values (M) of 5.90, 5.92 and 4.31, respectively, compared with pKi values of 9.31, 8.18 and 7.57 determined in the [3H]-DPCPX binding assay in rat cortical membranes. These novel pyrazolopyridine derivatives therefore represent high affinity, adenosine A1 receptor selective drugs that, in contrast to xanthine based antagonists, exhibit similar affinity for [3H]-DPCPX binding sites in human, rat, mouse and guinea-pig brain membranes. PMID- 9401788 TI - Effect of lacidipine on cholesterol esterification: in vivo and in vitro studies. AB - 1. Cholesterol esterification and accumulation in the arterial wall is a hallmark of atherogenesis. Several preclinical studies suggest that calcium antagonists may exert antiatherosclerotic activity by directly affecting atherogenesis in the arterial wall. We investigated the effect of the second generation dihydropyridine calcium antagonist lacidipine on cholesterol metabolism in vivo in the aortic arch of cholesterol fed rabbits, and in vitro in mouse cultured peritoneal macrophages. 2. Treatment of cholesterol-fed rabbits with 1, 3 and 10 mg kg-1 day-1 of lacidipine for two months reduced, in a dose-dependent manner, cholesterol esterification in the aortic arch: 24 +/- 6, 30 +/- 12, and 41 +/- 8% inhibition, respectively (P < 0.001 vs HC control). Concomitantly, drug treatment reduced total cholesterol content of the vessel wall. Lacidipine 3 and 10 mg kg-1 day-1 reduced cholesterolaemia (approximately 20%); no effect was observed at the lowest dose used (1 mg kg-1 day-1). These results suggest that the action of lacidipine on cholesterol esterification in the arterial wall involves, at least in part, a direct effect on cellular cholesterol metabolism. Inhibition of cholesterol esterification in the arterial wall was observed also in a reference group of animals treated with the specific ACAT inhibitor CI-976. 3. To evaluate the action of lacidipine on intracellular cholesterol metabolism we performed in vitro experiments with murine macrophages, the main cell type that accumulates cholesterol in the arterial wall. Lacidipine almost completely inhibited cholesterol esterification in cholesterol loaded macrophages in culture. The effect was observed independently of esterification stimuli and in cell free homogenates. The drug modified intracellular cholesterol distribution, doubling the free- to esterified sterol ratio, but did not influence the cellular rate of cholesteryl ester hydrolysis in the cell. All together these results indicate an inhibitory effect of lacidipine on cholesterol esterification catalyzed by the enzyme ACAT in murine macrophages. 4. We concluded that lacidipine influences cellular cholesterol metabolism. This effect may contribute to the potential antiatherosclerotic activity of this drug. PMID- 9401789 TI - Reversal of tolerance to the antitransit effects of morphine during acute intestinal inflammation in mice. AB - 1. The aim of investigation was to establish and compare the reversibility of tolerance to the antitransit effects of morphine by three different procedures: (a) acute inflammation of the gut, (b) lorglumide a cholecystokininA (CCKA) receptor antagonist, or (c) MK-801, an N-methyl-D-aspartate (NMDA) receptor ion channel blocker. The type of interaction between morphine and lorglumide or MK 801 on the inhibition of gastrointestinal transit (GIT) in naive animals was also evaluated. 2. Male Swiss CD-1 mice were implanted with 75 mg of morphine base or placebo pellets. Gastrointestinal transit was assessed with a charcoal meal and results expressed as % inhibition of GIT. Inflammation was induced by the intragastric (p.o.) administration of croton oil (CO), while controls received castor oil (CA) or saline (SS). Morphine was administered by subcutaneous (s.c.) or intracerebroventricular (i.c.v.) injection, to naive and tolerant animals treated with CO, CA or SS. Dose-response curves for s.c. morphine were also performed in naive and tolerant mice receiving 5.2 or 7.4 nmol (s.c.) lorglumide or MK-801, respectively. 3. The ED50 values for inhibition of GIT by s.c. morphine were: 45.9 +/- 2.7 and 250.1 +/- 3.1 nmol in naive and tolerant animals, respectively, demonstrating a five fold decrease in the potency of morphine. In naive animals, inflammation (CO) decreased the ED50 of morphine three times (14.4 +/- 2.2 nmol). However, no tolerance to s.c. morphine (ED50 16.4 +/- 2.6 nmol) was manifested during intestinal inflammation. After i.c.v. administration, a similar degree of tolerance to morphine was observed (4.8 fold decrease in potency). Intestinal inflammation had no effect on the ED50 values of i.c.v. morphine in naive and tolerant animals, showing that reversal of tolerance is related to local mechanism/s. Mean values for intestinal pH were 6.9 +/- 0.04 and 6.2 +/- 0.04 in SS and CO treated mice, respectively. In addition, morphine was 74 times more potent by the i.c.v. than by the s.c. route (naive-SS). 4. Morphine and lorglumide interacted synergistically in naive animals; in addition, the administration of lorglumide reversed tolerance to s.c. morphine. No interaction (additivity) was observed in naive animals when morphine and MK-801 were administered in combination. However, the drug completely reversed tolerance to the antitransit effects of morphine. 5. The present investigation shows that acute inflammation of the gut reverses tolerance to the antitransit effects of s.c. morphine by a peripheral mechanism. Qualitatively similar results were obtained after the administration of lorglumide or MK-801. Our results suggest that a local decrease in pH could play an important role during inflammation, while antagonism of endogenous compensatory systems would explain the reversal of tolerance induced by lorglumide or MK-801. PMID- 9401790 TI - Effects of a novel guanylate cyclase inhibitor on nitric oxide-dependent inhibitory neurotransmission in canine proximal colon. AB - 1. Previous studies suggested that nitric oxide (NO) may cause hyperpolarization and relaxation of canine colonic smooth muscle by both cGMP-dependent and cGMP independent mechanisms. This hypothesis was tested using 1H-[1,2,4]oxadiazolo[4,3 a]quinoxaline-1-one (ODQ), a novel inhibitor of NO-stimulated guanylate cyclase. 2. In the presence of histamine (30 microM), atropine and indomethacin (both at 1 microM), electrical field stimulation of intrinsic neurons (EFS; 5 Hz) produced inhibition of phasic contractile activity that is due to NO synthesis. ODQ caused a concentration-dependent block of this response (10 nM to 10 microM). 3. Inhibitory junction potentials (IJPs) due to NO synthesis were recorded from muscle cells located near the myenteric border of the circular muscle layer, using intracellular microelectrodes. IJPs were abolished by ODQ (1-10 microM). 4. EFS (10-20 Hz) produced frequency-dependent inhibition of electrical slow waves recorded from cells located near the submucosal surface of the circular muscle layer. This inhibition is due to NO synthesis, and it was abolished by ODQ (1-10 microM). 5. Hyperpolarization and relaxation produced by an NO donor, sodium nitroprusside, were abolished by ODQ pretreatment (1-10 microM). In contrast, inhibitory responses to 8-Br-cGMP (1 mM) were unaffected by ODQ. 6. ODQ alone (1 10 microM) had no significant effect on spontaneous electrical or phasic contractile activity. In tissues pre-treated with L-NAME (300 microM), ODQ decreased the amplitude of spontaneous or histamine-stimulated phasic contractile activity. 7. These results suggest that electrical and mechanical effects of endogenously released and exogenously applied NO in canine colon are largely due to cGMP synthesis by ODQ-sensitive soluble guanylate cyclase. No evidence to support a direct (cGMP-independent) mechanism of NO action was found. ODQ also appears to cause a non-specific inhibition of muscle contractile activity; however, this effect does not contribute to block of NO-dependent effects. PMID- 9401791 TI - Structural requirements for roxatidine in the stimulant effect of rat gastric mucin synthesis and the participation of nitric oxide in this mechanism. AB - 1. The structural requirements of the histamine H2-receptor antagonist, roxatidine (2-acetoxy-N-(3-[m-(1-piperidinylmethyl)phenoxy]-propyl)acetamide hydrochloride), for the stimulant effect on mucin biosynthesis and their relation to histamine H2-receptor antagonism were identified by considering the structural analogues of this drug using an organ culture system of the rat stomach and competition studies with [125I]iodoaminopotentidine ([125I]-APT) binding to membranes of the guinea pig striatum. 2. [3H]Glucosamine incorporation into mucin during 5 h incubation period was stimulated by roxatidine and its structural analogues A (2-hydroxy-N-(3-[m-(1-piperidinylmethyl)phenoxy]-propyl)acetamide) and B (N-(3-[m-(1-piperidinylmethyl)phenoxy]-propyl)acetamide). This effect was seen in mucosal cultures of the corpus, but not antrum, region. 3. Structural analogues, in which the length of the flexible chain between the benzene ring and the amide structure differs from that of roxatidine, failed to activate mucin synthesis. No significant change in mucus synthesis occurred with the addition of analogues in which the piperidine ring attached to the benzene ring via a methylene bridge was changed. 4. Specific [125I]-APT binding to the histamine H2 receptor of guinea pig brain membranes was inhibited by roxatidine and all structural analogues used in this study, except F (N-(3-[m-(N, N-dimethyl aminomethyl)phenoxy]-propyl)acetamide). 5. Ranitidine at 10(-4) M did not suppress the roxatidine-induced increase in [3H]glucosamine incorporation into mucin. 6. Roxatidine-induced stimulation of [3H]glucosamine incorporation into mucin was completely blocked by the addition of either NG-nitro-L-arginine (10( 5) M) or 2-(4-carboxyphenyl)-4,4,5,5,-tetramethylimidazoline-1-oxyl-3-oxide sodium salt (10(-5) M). The inhibitory action of NG-nitro-L-arginine was totally reversed by L-arginine (5 x 10(-3) M). 7. These results suggest that the cardinal chemical features of roxatidine for the activation of mucin biosynthesis in the corpus region of the rat stomach are the appropriate length of the flexible chain between the amide structure and the aromatic ring system bearing the methylpiperidinyl group at the meta position. The activity of roxatidine and its analogues to stimulate mucin synthesis is not related to their histamine H2 receptor antagonistic activity. Roxatidine-induced activation of mucin biosynthesis in the corpus tissue is mediated by nitric oxide. PMID- 9401792 TI - Acute effects of nitric oxide blockade with L-NAME on arterial haemodynamics in the rat. AB - 1. We employed the technique of impedance spectral analysis to investigate the role of endogenous nitric oxide (NO) in the regulation of steady and pulsatile haemodynamics in Wistar Kyoto rat (WKY). 2. A total of 12 WKYs was anaesthetized with pentobarbitol sodium (40 mg kg-1, i.p.) and artificially ventilated with an animal respirator. The aortic pressure wave was monitored with a high fidelity Millar sensor, and aortic flow wave with an electromagnetic flow probe. The pressure and flow waves were subjected to Fourier transform for the analysis of impedance spectra. 3. The baseline cardiovascular parameters were mean arterial pressure (APm) 95 +/- 9 mmHg, heart rate (HR) 338 +/- 9 b.p.m., stroke volume (SV) 0.23 +/- 0.01 ml, cardiac output (CO) 77.8 +/- 1.6 ml min-1, total peripheral resistance (TPR) 98 +/- 11 (x10(3)) dyne s cm-5, characteristic impedance (Zc) 2046 +/- 141 dyne s cm-5, arterial compliance at mean AP (Cm) 3.78 +/- 0.22 microliters mmHg-1 and backward pulse wave (Pb) 12.9 +/- 0.6 mmHg. 4. An NO synthase inhibitor, NG-nitro-L-arginine monomethyl ester (L-NAME) was administered at graded intravenous doses. This agent caused dose-dependent increases in AP and TPR with decreases in HR. At an accumulative dose of 10 mg kg 1, APm was increased by 29 +/- 3 mmHg (+31%) and TPR by 49 +/- 6 (x10(3)) dyne s cm-5 (+50%), while HR was reduced by 37 +/- 5 b.p.m. (-11%) and CO by 10.4 +/- 0.8 ml min-1 (-14%). The pulsatile haemodynamics including Zc and Pb were slightly increased by 14-15%. Cm was decreased by 1.09 microliters mmHg-1 (-29%). L-NAME also did not significantly affect the ventricular work including the steady, oscillatory and total work. 5. Aminoguanidine, a specific inhibitor for inducible NO synthase (iNOS), in dose 10-60 mg kg-1 i.v. did not alter the AP, HR and other parameters. The result indicated that blockade of constitutive NOS, but not iNOS is involved in these changes. 6. Angiotensin II (Ang) in various infusion doses was used to produce a profile of AP increase similar to that caused by L-NAME. Ang remarkably increased Zc, while TPR was moderately elevated. The pattern of haemodynamic changes was different from that following L-NAME. 7. The results suggest that blockade of the endogenous NO affects predominantly the arterial pressure and peripheral resistance. The Windkessel functions such as arterial impedance and pulse wave reflection are slightly increased. Ventricular works are not significantly altered. PMID- 9401793 TI - Lipolytic effects of conventional beta 3-adrenoceptor agonists and of CGP 12,177 in rat and human fat cells: preliminary pharmacological evidence for a putative beta 4-adrenoceptor. AB - 1. The nature of rat and human fat cell beta 3-adrenoceptors was investigated by studying the effects of the new beta 3-adrenoceptor selective antagonist, SR 59,230A, on lipolysis induced by the conventional beta 3-adrenoceptor agonists, CL 316,243 and SR 58,611A, and by the non-conventional partial beta 3 adrenoceptor agonist CGP 12,177 (a potent beta 1- and beta 2-adrenoceptor antagonist with partial beta 3-adrenoceptor agonist property). 2. In rat fat cells, the rank order of potency of agonists was: CL 316,243 > isoprenaline > SR 58,611A > CGP 12,177. The three former agents were full agonists whereas CGP 12,177 was a partial agonist (intrinsic activity of 0.70). In human fat cells, the lipolytic effect of CGP 12,177 reached 25% of isoprenaline effect. CL 316,243 was a poor inducer of lipolysis and SR 58,611A was ineffective. 3. In rat fat cells, lipolysis induced by CL 316,243 and SR 58,611A was competitively antagonized by SR 59,230A. Schild plots were linear with pA2 value of 6.89 and 6.37, respectively. Conversely, 0.1, 0.5 and 1 microM SR 59,230A did not modify the concentration-response curve of CGP 12,177. A rightward shift of the curve was however observed with 10 and 100 microM of SR 59,230A. The apparent pA2 value was 5.65. The non-selective beta-adrenergic antagonist, bupranolol, competitively displaced the concentration-response curve of CGP 12,177 and CL 316,243. Schild plots were linear with pA2 values of 6.70 and 7.59, respectively. CL316,243 mediated lipolytic effect was not antagonized by CGP 20,712A. In human fat cells, CGP 12,177-mediated lipolytic effect was antagonized by bupranolol and CGP 20,712A. SR 59,230A (0.1, 1 and 10 microM) did not modify the concentration response curve of CGP 12,177. A rightward shift was however observed at 100 microM leading to an apparent pA2 value of 4.32. 4. The results suggest that the non-conventional partial agonist CGP 12,177 can activate lipolysis in fat cells through the interaction with a beta-adrenoceptor pharmacologically distinct from the beta 3-adrenoceptor, i.e. through a putative beta 4-adrenoceptor. They suggest that the two subtypes coexist in rat fat cells whereas only the putative beta 4-adrenoceptor mediates lipolytic effect of CGP12,177 in human fat cells. PMID- 9401794 TI - Neuroprotective efficacy of ebselen, an anti-oxidant with anti-inflammatory actions, in a rodent model of permanent middle cerebral artery occlusion. AB - 1. The aim of this study was to investigate whether delayed treatment with the anti-oxidant and anti-inflammatory agent ebselen reduces the volume of infarction in a rodent model of permanent focal cerebral ischaemia. 2. Ebselen (10 or 30 mg kg-1) or vehicle was administered by gavage 30 min and 12 h after the induction of cerebral ischaemia by permanent occlusion of the left middle cerebral artery (MCA). Animals were killed 24 h following MCA occlusion, and the volumes of ischaemic damage in the ebselen and control groups were evaluated by quantitative histopathology. 3. Ebselen was quickly absorbed following oral (gavage) administration and reached peak levels in the plasma by 1 h post-administration (plasma selenium level of 0.68 +/- 0.04 and 0.84 +/- 0.1 microgram ml-1 for 10 and 30 mg kg-1, respectively, compared to control level of 0.51 +/- 0.02 microgram kg-1). 4. Treatment with the lower dose of ebselen (10 mg kg-1) significantly (P < 0.01) reduced the volume of infarction in the cerebral hemisphere and cerebral cortex (by 31.8% and 36.7%, respectively compared with the placebo group). 5. The neuroprotective efficacy of the higher dose ebselen (30 mg kg-1) was less than that of the lower dose ebselen (10 mg kg-1). The volume of ischaemic damage in the cerebral hemisphere was reduced by 23.7% (P < 0.02), and cerebral cortex by 27.5% (P < 0.01). 6. Both doses of ebselen (10, 30 mg kg-1) had no therapeutic efficacy on the caudate nucleus, where ischaemia was most severe, in this model. 7. Free radical-mediated injury is normally associated with reperfusion of ischaemic tissue. The present results suggest that oxidative injury is also a significant contributor to brain damage in models of maintained (permanent) ischaemia and that ebselen is effective in attenuating this free radical-induced damage. PMID- 9401795 TI - Variabilities in the distribution of neurofibrillary tangles in the anterior parahippocampal gyrus at initial stages of Alzheimer's disease. AB - The entorhinal region is located in the ventromedial surface of the temporal lobe on the parahippocampal gyrus. Occurrence of early argyrophilic neurofibrillary tangles was investigated in brain sections of the left and right entorhinal region of 9 individuals of both sexes between the age of 26 to 54 years. Parasagittal serial sections at 100 microns were performed to include the very anterior part of the entorhinal region. An advanced silver method was used for staining neurofibrillary changes. The neurofibrillary tangles were counted via light microscopy, marked on the sections and staged as proposed by Braak and Braak. In 1 case we found a left-right difference by 1 stage (right: stage I, left: stage II). The tangle distribution in another case exhibited several neurofibrillary tangles only in the very anterior part of the entorhinal cortex on both sides (stage 0). In 3 cases some neurofibrillary tangles occurred in the very anterior part of the right entorhinal region, while a few are present in more posterior parts (stage I). The corresponding left hemisphere showed the same anterior-posterior distribution pattern as on the right in only 1 of these cases. In the remaining 4 cases, no neurofibrillary changes were visible. In conclusion, the very anterior part of the entorhinal region is among the earliest sites for development of neurofibrillary tangles. Distribution of early Alzheimer-related neurofibrillary changes in the entorhinal region displays variabilities in the anterior-posterior direction as well as in a left-right comparison. PMID- 9401796 TI - Acute stroke in a child with miliary tuberculosis. AB - Neurological symptoms in childhood miliary tuberculosis are generally caused by underlying tuberculous meningitis (TBM), since the 2 conditions commonly occur concurrently. Cerebral infarction, a well-recognized complication of TBM, usually results from tuberculous periarteritis and secondary thrombosis. Neuropathological studies have demonstrated that the anterior cerebral circulation is more commonly affected than the arteries of the vertebro-basilar system, and basilar artery occlusion as a presenting manifestation of childhood miliary tuberculosis or TBM has not been described before. We report a 13-month old infant who presented with fever and convulsions, terminating in acute decerebration after a second prolonged seizure 1 week after the onset of symptoms. Magnetic resonance (MR) imaging demonstrated density changes compatible with acute vertebro-basilar ischemia as well as multiple cerebral granulomas. A chest radiograph showed diffuse miliary tuberculosis. Postmortem examination confirmed this diagnosis and revealed acute occlusion of the basilar artery by an infected (septic) thromboembolus showing granulomatous inflammation, which most likely arose from an endocardial vegetation with identical histology. PMID- 9401797 TI - Leptomeningeal lipomatous hamartoma overlying a midline cleft of the ventral pons. AB - Intracranial lipomatous hamartomas (lipomas) are of maldevelopmental nature and have a predilection for the midsagittal plane. They are often associated with malformations of the CNS and other organ systems. It is reported an asymptomatic lipomatous hamartoma of the ventral pontine leptomeninges and a midline cleft of the subjacent pons discovered incidentally at autopsy of an 80-year-old man. Review of the literature shows that this type of lesion at the ventral pons has not been reported previously. PMID- 9401799 TI - Light-microscopic study of the beta 1 integrin subunit in human skeletal muscle. AB - The beta 1 integrin subunit is identical with the CD29 antigen, which is found at the surface of leukocytes. Integrins are involved in cell-cell and cell-matrix adhesion, mediate neuronal attachment and neurite outgrowth in response to extracellular matrix proteins in cell culture systems. A few analyses of beta 1 integrin subunit have been done on developing and regenerating skeletal muscle in animals; but cell culture systems and animal models differ in some respects from human skeletal muscle in situ. The expression of a beta 1 integrin subunit variant in human skeletal muscle was reported merely by Western blot analysis. Our present study, performed with immunohistochemical procedures, attempts to demonstrate the expression of the beta 1 integrin subunit in developing, normal adult, and diseased human skeletal muscles. The results demonstrated that the beta 1 integrin subunit is expressed in developing, normal adult, regenerating, and denervated human skeletal muscle. In developing muscle, the beta 1 integrin subunit was observed in muscle cells at least from 12 to 16 weeks of gestation. In muscular dystrophy and inflammatory myopathy the beta 1 integrin subunit staining occurs in basophilic muscle fibers. Furthermore, the beta 1 integrin subunit is expressed in mature fast twitch type 2 fibers, and in denervated myocytes in neurogenic muscular atrophy. On serial sections, the beta 1 integrin subunit, N-CAM (neural cell adhesion molecule) and vimentin are expressed in identical muscle fibers. However, in mature fast twitch type 2 fibers the beta 1 integrin subunit is expressed exclusively and in neurogenic muscular atrophy vimentin expression is weak. In conclusion, the beta 1 integrin subunit, in human skeletal muscles, probably plays a role in the growth morphology and innervation of developing, regenerating, and denervated myocytes. Furthermore, the observation that the beta 1 integrin subunit is enriched in mature fast twitch type 2 fibers indicates that the beta 1 integrin subunits may play a role in transducing mechanical forces to extracellular matrix proteins. PMID- 9401800 TI - Pure alexia: clinical-pathologic evidence for a lateralized visual language association cortex. AB - Traditional views of pure alexia have held that the disorder results from a disconnection between the secondary visual cortices of both hemispheres and the angular gyrus of the dominant hemisphere. Evidence has accumulated, however, suggesting the importance of the posterior inferior temporal area in visual language processing. We describe clinical-pathological support for the presence of a lateralized visual language association area residing in the dominant posterior inferior temporal lobe. PMID- 9401798 TI - Adhesion molecules in HIV-related and idiopathic polymyositis: immunohistochemical studies. AB - Idiopathic polymyositis (IPM) and HIV polymyositis (HIV-PM) are considered to be related autoimmune diseases whose target is skeletal muscle. They have been associated to a T cell-mediated and MHC-I-restricted cytotoxic phenomenon, but both etiology and physiopathology remain incompletely understood. Their histological hallmarks are mononuclear leukocyte infiltrates as well as necrosis, degeneration, and regeneration of muscle fibers. In the present study, we have investigated the immunohistochemical expression of cell adhesion molecules, cytokines, and leukocyte surface antigens in biopsies of HIV-PM and IPM patients. The aim was to better define factors involved in lymphocyte recruitment and in inflammatory changes seen in PM. Notable upregulation of ICAM-1 and TNF-alpha was detected on capillary and venular endothelia and on inflammatory cells, whereas no significant VCAM-1 and ELAM-1 expression was present. LFA-1, the main ICAM-1 counter-receptor, was found to be highly expressed on lymphocytes and monocytes, especially at the vicinity of damaged fibers. The majority of infiltrating cells were CD8+CD45 RO-T cells, which are thought to have memory capacities. These findings suggest that in IPM and HIV-PM, enhanced ICAM-1 and LFA-1 expression possibly induced by TNF-alpha, may regulate the homing process of selected lymphocyte clones in muscle tissue. Lymphocyte proliferation and differentiation into memory subsets may further potentiate tissue-restricted homing capabilities. PMID- 9401801 TI - p53 status has no prognostic significance in glioblastomas treated with radiotherapy. AB - Mutation of the tumor suppressor gene TP53 and accumulation of the p53 protein are common events in the range of cerebral astrocytic tumors, including glioblastomas, but it is uncertain whether these events are associated with prognosis. Evidence suggests that the response of various tumors to radiotherapy may be influenced by the status of p53 which is involved in control of the cell cycle and apoptosis. This study tests the hypothesis that p53 governs survival in patients (n = 62) with glioblastomas who have received radiotherapy. Analysis of TP53 and p53 immunohistochemistry were undertaken using standard methods. TP53 mutations were present in 27% tumors, while 50% were p53-immunopositive (LI > 3%). A strong correlation (p < 0.0001) was found between a high p53 LI (> 50%) and the presence of a mutation, but p53 status at the level of gene or protein was unrelated to survival. Radiation-induced apoptosis that is independent of p53, and the presence in glioblastomas of genetic abnormalities that are also involved in the cellular response to radiation, such as deletions and mutations of pRb, are possible explanations of this result. PMID- 9401802 TI - Malignant melanoma in the CNS, subtyping and immunocytochemistry. AB - Paraffin-embedded specimens from 21 patients (mean age 49 years) with malignant melanocytic tumors of the central nervous system were studied. Extraneuronal primary tumors were situated at the trunk (38%), the lower (14%) or upper extremity (10%), and the head/neck region (5%). In 33% no extraneural primary tumor could be detected. The tumor location was frontal (19%), occipital (19%), parietal, spinal, multifocally (14%, respectively), or temporal (5%). Four subtypes were distinguished according to the predominant histological cell type: pleomorphic, epithelioid, spindle- and mixed-cell tumors. 29% contained no melanin, most of them belonging to the epithelioid subtype. The morphology and immunohistochemical reactivity for different antibodies (KL-1, EMA, VIM, HMB-45, NKI-C3, S-100, and MIB-1/Ki-67) were assessed. Positive staining was demonstrated for HMB-45 (in 86% of cases), NKI-C3 (100%), S-100 (95%), vimentin (75%), and KL 1 (33%). No expression of the cytokeratin EMA could be detected. The mean proliferation index measured by MIB-1 immunoreactivity was 21%. The 4 histological subtypes were found to express different antigen patterns. In the analysis of CNS tumors of unknown origin, the panel of antibodies used for diagnosis should include HMB-45 as the most specific marker for malignant melanoma. PMID- 9401803 TI - Polymerase chain reaction for the detection of Burkholderia pseudomallei. AB - Melioidosis is a potentially lethal infection of humans and animals in Southeast Asia and northern Australia. Current methods for detection of the causative organism, Burkholderia pseudomallei, lack both speed and sensitivity. We report the development of a highly sensitive polymerase chain reaction-based method that can detect as few as 35 colony-forming units of B. pseudomallei/mL in saline suspensions. This polymerase chain reaction test also detected the presence of B. pseudomallei DNA in culture-negative splenic tissue obtained from mice infected with the organism, but without clinical evidence of disease. Specificity has been confirmed using a variety of pathogenic and nonpathogenic organisms, including B. mallei, B. cepacia, and Pseudomonas species. The clinical usefulness of this test should be assessed prospectively and compared with conventional diagnostic techniques. PMID- 9401804 TI - Transferable antibiotic resistance in nosocomial Stenotrophomonas maltophilia strain. AB - Stenotrophomonas maltophilia (298/85) was isolated from the extensively inflamed conjunctiva of a neonate in a regional hospital in Ostrava, Czech Republic. It was resistant to all available antibiotics except cefepime and trimethoprim. The donor S. maltophilia strain 298/85 transferred carbenicillin and cephaloridine resistance determinants to recipient strains of Escherichia coli K-12 3110 rif+ and Proteus mirabilis P-38 rif+. All transconjugant colonies were co-resistant also to kanamycin, cefotaxime, and aztreonam. Active hydrolysis of imipenem in the original strain was inhibited by ethylene diamine tetra-acetic acid, and hydrolysis of cefotaxime and aztreonam in the original strain and in the E. coli K-12 transconjugant was inhibited by clavulanate. In contrast, ceftazidime was hydrolyzed by the original strain and was not inhibited by clavulanate, indicating a different character of the resistance to cefotaxime or aztreonam and ceftazidime. PMID- 9401805 TI - Ciprofloxacin-resistant Escherichia coli emerging in a rehabilitation medical center. AB - A retrospective review of laboratory records from 1988 to 1996 has shown an increased rate of ciprofloxacin-resistant (cip(r)) Escherichia coli in our rehabilitation center. Resistance increased from 0.6% in 1989 to 5.9% in 1996. Of 7870 E. coli strains isolated during this period, 257 cip(r)-E. coli were recovered from 257 patients. The majority (96%) of these resistant strains were isolated from the urine samples. One hundred and twenty strains of cip(r)-E. coli were also resistant to four other fluoroquinolones. MICs ranging from 64 to 512 micrograms/mL were observed in 75% of the strains and > or = 1028 micrograms/mL in 6.4% of the strains. Resistance to ciprofloxacin was due to possible mutations in topoisomerase gyrA. PMID- 9401806 TI - A comparison of the BioStar Strep A OIA rapid antigen assay, group A Selective Strep Agar (ssA), and Todd-Hewitt broth cultures for the detection of group A Streptococcus in an outpatient family practice setting. AB - In some studies the BioStar Strep A OIA (optical immunoassay) has yielded inconsistent results, although originally it was reported to be more sensitive than conventional culture for the detection of group A Streptococcus (GAS). The Group A Selective Strep Agar with 5% sheep blood (ssA) incubated anaerobically has been reported to be more sensitive than conventional culture in the detection of GAS. We compared the BioStar Strep A OIA GAS rapid antigen detection kit to anaerobic culture on ssA with and without preincubation in Todd-Hewitt broth (THB) for the detection of GAS. From September 1995 through January 1996, throat swabs were collected in duplicate from 75 children (< or = 18 years) and 188 adults (> 18 years) who presented with pharyngitis in the outpatient University of New Mexico Family Practice Clinic. Thirty-one (12%) of the 263 cases were positive for GAS by culture and/or broth. Compared with anaerobic culture on the ssA, with and without preincubation in THB, the sensitivity, specificity, positive predictive value, and negative predictive value of the BioStar Strep A OIA were 77, 62, 22, and 95%, respectively. Compared with enrichment in THB followed by subculture on ssA and anaerobic incubation, the sensitivity, specificity, positive predictive value, and negative predictive value of direct culture on ssA and anaerobic incubation were 79, 99, 98, and 96%, respectively. All isolates were serologically grouped. The BioStar Strep A OIA is as sensitive as direct culture on ssA incubated anaerobically, but the low specificity and low positive predictive value when the OIA is used in low prevalence populations could lead to unnecessary antibiotic treatment. PMID- 9401807 TI - An outbreak of Candida parapsilosis prosthetic valve endocarditis. AB - Candida parapsilosis, an important nosocomial pathogen and the most common species of Candida found on the hands of health care workers, is a rare cause of prosthetic valve endocarditis (PVE). From March through June 1994, four cases of C. parapsilosis PVE were diagnosed at a 400-bed community hospital. The mean time to presentation after valve replacement surgery was 148 days (range, 20 to 345). Three of the four patients died of complications of PVE. Multiple environmental cultures were performed, and only one was positive for C. parapsilosis. Cultures from the bypass pump, cell saver, cardioplegia solution, and subsequent valves were all negative. All valve replacements were performed by the same operating room team. Interviews with the surgeon and physician assistant, the only personnel involved in all cases, revealed that their hypoallergenic gloves were subject to frequent tears during valve replacement procedures, often requiring several glove changes per procedure. Hand cultures of personnel were obtained, and cultures from 20 individuals (26%) were positive for C. parapsilosis. Hand cultures of the surgeon and physician assistant obtained 8 months after the last case had surgery were negative for yeasts. Molecular typing of the 3 available case isolates, 14 epidemiologically unrelated patient isolates, 1 environmental isolate, and 20 hand isolates was performed by electrophoretic karyotyping and restriction endonuclease analysis of genomic DNA using restriction enzymes BssHII and EagI followed by pulsed field gel electrophoresis. The three case isolates were identical by restriction endonuclease analysis of genomic DNA, and two of the three shared the same electrophoretic karyotyping profile. The remaining patient, environmental, and hand isolates represented 29 different DNA types and were distinctly different from the case isolates. All of the isolates tested were susceptible to amphotericin B, 5FC, fluconazole, and itraconazole. The circumstantial evidence suggests the probability of glove tears during valve replacement surgery and subsequent transmission of C. parapsilosis to patients. PMID- 9401808 TI - A double-blind, randomized study of three antimicrobial regimens in the prevention of infections after elective colorectal surgery. AB - The objective of this study was to assess the prophylactic efficacy of cefoxitin, ceftizoxime, and metronidazole-gentamicin in colorectal surgery. A double-blind, randomized prospective clinical trial design was used in a Canadian tertiary care teaching hospital. Patients were randomized to one of three treatment groups and received three doses of a study drug (30 min preoperative and 2 postoperative doses at 12 and 24 h). Cefoxitin and ceftizoxime were given as 1000-mg doses. Metronidazole-gentamicin was given as 500 mg of metronidazole plus 120 mg of gentamicin in a minibag. High-risk patients (bowel ischemia, diabetic, current steroid use, etc.) received 10 postoperative doses. Patients with infections, prior antibiotics, or study drug allergies were excluded. Over 30 months, 153 patients were enrolled. Thirty-one patients were excluded for protocol violations. Of the 122 evaluable patients (38 ceftizoxime, 45 metronidazole gentamicin, 39 cefoxitin), there was no difference across groups regarding sex, age, weight, preoperative Apache II score, and prior history of bowel surgery. Groups were equivalent regarding surgeon, nursing unit, high-risk status (six ceftizoxime, seven metronidazole-gentamicin, seven cefoxitin), bowel preparation, and procedure (including blood loss, drains, organ injury, intraoperative complications). Clinically significant infection requiring systemic antibiotics (7-day hospital and 30-day follow-up) was identified in 0% of ceftizoxime, 15% of metronidazole-gentamicin, and 26% of cefoxitin receiving patients (p = 0.005). Mean ASEPSIS scores for each group were 2.3 (range 0-15) for ceftizoxime, 9.2 (range 0-45) for metronidazole-gentamicin, and 10.4 (range 0-75) for cefoxitin (p = 0.01). Ceftizoxime patients tended to have a shorter total hospital stay (12.2 days versus 19.7 days for cefoxitin versus 13.9 days for metronidazole gentamicin; p = 0.04), although the procedure to discharge interval was not significantly different (p = 0.09). There was no difference in clinical outcome according to risk status. Anaerobic bacteria were observed more commonly in the ceftizoxime and cefoxitin groups, whereas enteric Gram-negative aerobes were observed most often in the metronidazole-gentamicin group. The study regimens were generally well tolerated. Drug costs were equivalent between ceftizoxime and cefoxitin and lowest with the metronidazole-gentamicin regimen. Ceftizoxime appears to be more effective for the prevention of infection in colorectal surgery than either cefoxitin or metronidazole-gentamicin in the dosage regimens studied. PMID- 9401809 TI - In vitro activity of clarithromycin alone and in combination with ciprofloxacin or levofloxacin against Legionella spp.: enhanced effect by the addition of the metabolite 14-hydroxy clarithromycin. AB - Clarithromycin is metabolized to an active metabolite, 14-hydroxy clarithromycin. These compounds have demonstrated excellent in vitro activity against Legionella species, with both agents having significantly lower MICs than erythromycin. Using a checkerboard assay, the activity of clarithromycin and its hydroxy metabolite, alone and in combination, was examined against 41 Legionella organisms. The activity of clarithromycin and 14-hydroxy clarithromycin, in a 2:1 ratio, plus ciprofloxacin or levofloxacin was also determined. Activity of the antibiotic combinations was determined by calculating the fractional inhibitory concentration index. An agar dilution method using buffered charcoal yeast extract media was used for susceptibility and synergy testing. An inoculum of 10(4) CFU/spot was used, with all plates incubated at 35 degrees C for 48 h. The MIC90 for clarithromycin or 14-hydroxy clarithromycin alone was 0.5, versus 0.25 microgram/mL for the combination. Additive effects were observed with clarithromycin and its hydroxy metabolite for 61% of the Legionella species, with fractional inhibitory concentration indices ranging from 0.63 to 1.25. The 14 hydroxy metabolite significantly increased the activity of both fluoroquinolone/clarithromycin combinations. Based on these data, in vitro susceptibility testing of agents such as clarithromycin should be reevaluated to account for the activity of active metabolites. PMID- 9401810 TI - Comparative in vitro assessment of sparfloxacin activity and spectrum using results from over 14,000 pathogens isolated at 190 medical centers in the USA. SPAR Study Group. AB - Sparfloxacin, a new orally administered fluoroquinolone, was tested against 14,182 clinical strains isolated (generally blood stream and respiratory tract cultures) at nearly 200 hospitals in the United States (USA) and Canada. Sparfloxacin activity was compared with 13 other compounds by Etest (AB BIODISK, Solna, Sweden), broth microdilution, or a standardized disk diffusion method. Using the Food and Drug Administration/product package insert MIC breakpoint for sparfloxacin susceptibility (< or = 0.5 microgram/ml), 94% of Streptococcus pneumoniae (2666 isolates) and 89% of the other streptococci (554 isolates) were susceptible. However, at < or = 1 microgram/ml (the breakpoint for all nonstreptococcal species) sparfloxacin susceptibility rates increased to 100% and 98%, respectively, for the two groups of streptococci. Only 50% and 65% of pneumococci were susceptible to ciprofloxacin (MIC90, 3 micrograms/ml) and penicillin (MIC90, 1.5 micrograms/ml), respectively. Although there were significant differences between regions in the USA in the frequency of penicillin resistant pneumococcal strains, results indicate that the overall sparfloxacin MIC90 was uniformly at 0.5 microgram/ml. Nearly all (> or = 99%) Haemophilus species and Moraxella catarrhalis, including those harboring beta-lactamases, were susceptible to sparfloxacin, ciprofloxacin, and amoxicillin/clavulanic acid. Only cefprozil and macrolides demonstrated lower potency and spectrum against these two species. Sparfloxacin was active against oxacillin-susceptible Staphylococcus aureus (96 to 97%), Klebsiella spp. (95%), and other tested enteric bacilli (93%). Comparison between broth microdilution MIC and disk diffusion interpretive results for M. catarrhalis, Staphylococcus aureus, and the Enterobacteriaceae showed an absolute intermethod categorical agreement of > 95% using current sparfloxacin breakpoints, in contrast to those of cefpodoxime for S. aureus where a conspicuous discord (98% versus 59%) between methods was discovered. These results demonstrate that sparfloxacin possesses sufficient in vitro activity and spectrum versus pathogens that cause respiratory tract infections (indications), especially strains resistant to other drug classes such as the earlier fluoroquinolones, oral cephalosporins, macrolides, and amoxicillin/clavulanic acid. The sparfloxacin susceptibility breakpoint for streptococci may require modification (< or = 1 microgram/ml) based on the MIC population analysis presented here. A modal MIC (0.38 to 0.5 microgram/ml) was observed at the current breakpoint. Regardless, sparfloxacin inhibited 89% (nonpneumococcal Streptococcus spp.) to 100% (Haemophilus spp., M. catarrhalis) of the isolates tested with a median activity of 97% against indicated species. PMID- 9401811 TI - Antimicrobial activity of 12 broad-spectrum agents tested against 270 nosocomial blood stream infection isolates caused by non-enteric gram-negative bacilli: occurrence of resistance, molecular epidemiology, and screening for metallo enzymes. AB - A total of 270 recent nosocomial blood stream isolates of non-Enterobacteriaceae Gram-negative bacilli representing nearly 50 U.S. medical centers were characterized. The numbers of isolates of individual organisms were: Pseudomonas aeruginosa (n = 204), Acinetobacter spp. (n = 48), and Stenotrophomonas maltophilia (n = 18). MICs were determined using the broth microdilution susceptibility method with 12 antimicrobial agents: piperacillin, piperacillin/tazobactam, ceftriaxone, ceftazidime, cefepime, imipenem, ciprofloxacin, ofloxacin, amikacin, gentamicin, tobramycin, and trimethoprim/sulfamethoxazole. Based on current National Committee for Clinical Laboratory Standards breakpoints, rates of resistance to cefepime, ceftazidime, and imipenem were as follows: P. aeruginosa, 3, 9, and 5%; Acinetobacter spp., 2, 37, and 0%; and S. maltophilia, 88.7, 35.3, and 100%, respectively. Trimethoprim/sulfamethoxazole was the most active agent against S. maltophilia (100% susceptible). Twenty-eight isolates of P. aeruginosa that expressed high levels of resistance to ceftazidime (MIC, > 256 micrograms/mL) and imipenem (MIC, > 32 micrograms/mL) were examined for potential metallo-beta-lactamase production by polymerase chain reaction and were found to be negative. Molecular typing of P. aeruginosa isolates revealed many patient-unique strains, but also noted clustering of infections due to isolates of the same DNA type, suggesting possible nosocomial transmission in 9 of 14 medical centers. Given the resistance profile and pathogenic potential of these non-enteric Gram-negative bacilli, considerable effort should be exerted to develop and enforce infection control and antimicrobial utilization practices that will limit the spread of these organisms in the hospital environment. PMID- 9401812 TI - Intracerebral mass lesions in patients with human immunodeficiency virus infection and cryptococcal meningitis. AB - The frequency of intracerebral mass lesions (ICML) in patients with human immunodeficiency virus (HIV) infection and cryptococcal meningitis (CM) is not well established. Cryptococcoma seems to be a rare affliction. The objective of this study was to analyze the etiology of ICML in patients with HIV infection and CM. The methodology was a retrospective review of cases diagnosed in two Spanish hospitals between September 1988 and April 1995. Eighteen cases of CM were identified. Computed tomography was performed on presentation in 17 cases. Only one patient had ICML, which progressed while on antifungal treatment and regressed when anti-Toxoplasma treatment was established. During follow-up, two additional patients developed ICML and were successfully treated as toxoplasmosis. Overall, 3 out of 17 patients (18%) developed ICML and all three were cured when anti-Toxoplasma treatment was implemented. In our study, cerebral toxoplasmosis was the only presumed cause of ICML. In areas of high prevalence of toxoplasmosis, ICML in patients with CM may not be cryptococcomas. Consequently, in these areas of high prevalence, a trial of toxo-therapy should be strongly considered for patients with CM and ICML. PMID- 9401813 TI - Comparative antistreptococcal activity of two newer fluoroquinolones, levofloxacin and sparfloxacin. AB - The objective of this study was to evaluate the in vitro activity of sparfloxacin, levofloxacin, ofloxacin, and ciprofloxacin against contemporary strains of streptococci. Susceptibility testing of a panel of 300 recent clinical isolates of streptococci (100 each of beta-hemolytic, viridans group, and Streptococcus pneumoniae) using reference broth microdilution methods was performed, and the results were compared. Sparfloxacin was the most active of the four tested fluoroquinolones, inhibiting 99-100% of all isolates at concentrations of < or = 1 microgram/ml, and was two- to eightfold more potent than the three comparative agents. Levofloxacin was also quite active, inhibiting 98-100% of the isolates at concentrations < or = 2 micrograms/ml. Both sparfloxacin and levofloxacin possess an improved spectrum and potency against contemporary strains of streptococci compared to currently available fluoroquinolones. PMID- 9401814 TI - In vitro activity of chloramphenicol alone and in combination with vancomycin, ampicillin, or RP 59500 (quinupristin/dalfopristin) against vancomycin-resistant enterococci. AB - Using a checkerboard assay, ampicillin, vancomycin, and RP 59500, each in combination with chloramphenicol, were tested for synergy against 23 isolates of vancomycin-resistant enterococci. Additive effects were seen in 62.5% of the isolates when exposed to chloramphenicol plus RP 59500. Additive effects were observed in 20% and 15% of isolates with chloramphenicol plus vancomycin or ampicillin, respectively. No antagonism was noted. PMID- 9401816 TI - Protection of rat myocardium by mitogenic and non-mitogenic fibroblast growth factor during post-ischemic reperfusion. AB - The effects of acidic fibroblast growth factor (FGF-1) and basic fibroblast growth factor (FGF-2) and a non mitogenic form of FGF1 on myocardial ischemia and reperfusion were assessed. Rats underwent 10 minutes of coronary artery occlusion followed by 24 hours of reperfusion. Creatinine kinase content of the affected myocardium showed that both fibroblast growth factors 1 and 2 effectively protected against ischemia reperfusion injury (p < 0.01), and that the vasoactive but nonmitogenic form of the FGF1 was equally protective (p < 0.01 versus control + vehicle). The results were confirmed by light and electron-microscopy histological studies. Histological evaluations after treatment with the non mitogenic fibroblast growth factor 1 showed that it did not generate the severe hyperplasia and connective tissue disorganization observed with the native mitogenic proteins. The possibility of using a non-mitogenic form of fibroblast growth factor for cardio-protection circumvents many of the potentially undesirable effects that may derive from systemically introducing broad spectrum acting fibroblast growth factors in vivo. This myocardial protection observed 24 hours after the treatment with fibroblast growth factors, and the efficacy of the non-mitogenic form of the protein, also suggest that the protective effect of fibroblast growth factors may be due to the increased blood flow rather than to angiogenesis. PMID- 9401817 TI - Low level expression of basic FGF upregulates Bcl-2 and delays apoptosis, but high intracellular levels are required to induce transformation in NIH 3T3 cells. AB - We investigated the roles of basic fibroblast growth factor (bFGF) in the transformation and survival of NIH 3T3 cells. We constructed NIH 3T3-derived cell lines expressing human bFGF using retroviral gene transfer with an N2-based vector. Clonally derived cell lines containing a single copy of the vector overexpress bFGF mRNA and produce more immunoreactive protein (0.407 +/- 0.010 3.028 +/- 0.087 ng bFGF/10(6) cells) which is biologically active than nontransduced (0.151 +/- 0.013 ng bFGF/10(6) cells) or N2-transduced (0.211 +/- 0.029 ng bFGF/10(6) cells) NIH 3T3 cells. All cells producing excess amounts of bFGF achieve greater density at confluence, show delayed apoptosis and increased survival and have elevated intracellular levels of Bcl-2. However, only cells expressing from 8-15 times background levels of bFGF are phenotypically transformed. The transformed cells form dense foci at confluence, have decreased adherence to tissue culture plates and grow colonies in soft agar. Exogenous bFGF induces higher Bcl-2 levels in a dose dependent manner and recapitulates the antiapoptotic effects of the overexpressed species but fails to induce changes associated with the transformed phenotype. In this study, we demonstrate a dissociation between phenotypic transformation secondary to bFGF overexpression and upregulation of cellular Bcl-2 that correlates with a delay in programmed cell death. Although low level expression of bFGF or exogenous bFGF is sufficient to upregulate Bcl-2 and delay apoptosis, high intracellular levels are required for cellular transformation. These data suggest that overexpression of bFGF modulates cellular transformation and Bcl-2-mediated inhibition of apoptosis through alternate molecular mechanisms. PMID- 9401815 TI - Influence of PDGF-BB on proliferation and transition through the MyoD-myogenin MEF2A expression program during myogenesis in mouse C2 myoblasts. AB - We have previously demonstrated that PDGF-BB enhances proliferation of C2 myoblasts. This has led us to examine whether the mitogenic influence of PDGF-BB in the C2 model correlates with modulation of specific steps associated with myogenic differentiation. C2 myoblasts transiting through these differentiation specific steps were monitored via immunocytochemistry. We show that the influence of PDGF on enhancing cell proliferation correlates with a delay in the emergence of cells positive for sarcomeric myosin. We further monitored the influence of PDGF-BB on differentiation steps preceding the emergence of myosin+ cells. We demonstrate that mononucleated C2 cells first express MyoD (MyoD+/myogenin- cells) and subsequently, myogenin. Cells negative for both MyoD and myogenin (the phenotype preceding the MyoD+ state) were present at all times in culture and comprised the majority, if not all, of the cells which responded mitogenically to PDGF. Additionally, the frequency of the MyoD+/myogenin+ cell phenotype was reduced in cultures receiving PDGF, suggesting that PDGF can modulate the transition of the cells into the myogenin+ state. We determined that many of the myogenin+ cells subsequently become MEF2A+ and this phenomenon is not influenced by PDGF-BB. FGF-2 also enhanced the proliferation of C2 myoblasts and suppressed the appearance of the myogenin+ cells, but did not influence the subsequent transition into the MEF2A+ state. The study raises the possibility that PDGF-BB and FGF-2 might delay the transition of the C2 cells into the MyoD+/myogenin+ state by depressing a paracrine signal that enhances differentiation. PMID- 9401818 TI - Controlled release of leukaemia inhibitory factor (LIF) to tissues. AB - Leukaemia inhibitory factor (LIF) has been shown to effectively enhance skeletal muscle regeneration after mechanical injury and it may have potential therapeutic use in the muscular dystrophies as well as peripheral nerve repair after injury. When LIF is applied systemically to an animal, it is rapidly removed with a biological half life of only a few minutes, and at high doses it exhibits toxic effects. Calcium alginate rods have been developed for the purpose of insertion adjacent to skeletal muscles. These rods, when charged with LIF will release the growth factor to the muscle at a rate of less than 1% per day and for a period extending to several months. In addition, tubes of alginate are described which will be suitable for the continuous supply of LIF to repaired peripheral nerve. PMID- 9401819 TI - Placenta growth factor and vascular endothelial growth factor are co-expressed during early embryonic development. AB - We have used the polymerase chain reaction to identify mouse proteins similar in primary structure to the endothelial cell mitogen Vascular Endothelial Growth Factor (VEGF). One amplified product encoded mouse Placenta Growth Factor (PIGF). The pattern of PIGF gene expression in mouse embryos was studied by in situ hybridization. Transcripts encoding mouse PIGF were abundant in trophoblastic giant cells associated with the parietal yolk sac at early stages of embryogenesis. VEGF transcripts were also detected in trophoblastic giant cells raising the possibility that these cells may secrete heterodimers consisting of one PIGF subunit and one VEGF subunit. The secretion of PIGF and VEGF by trophoblastic giant cells is likely to be the signal which initiates and co ordinates vascularization in the deciduum and placenta during early embryogenesis. PMID- 9401820 TI - Regulation of gamma-glutamyl transpeptidase activity by Ca(2+)- and protein kinase C-dependent pathways in Sertoli cells. AB - gamma-Glutamyl transpeptidase (gamma-GTP) activity in Sertoli cells can be stimulated by FSH. This cAMP-dependent metabolic event can be enhanced when Sertoli cells are co-cultured with germ cells, suggesting that different signal transduction pathways may be involved in the regulation of gamma-GTP activity. In this study we examined the participation of Ca(2+)- and protein kinase C (pkC) dependent signal transduction pathways in the regulation of basal and FSH stimulated gamma-GTP activity. Under basal conditions, the increase in extracellular Ca2+ concentration or the addition of the Ca2+ ionophore 4Br-A23187 produced a decrease in gamma-GTP activity. Conversely, blockage of voltage dependent Ca2+ channels with verapamil or nifedipine or inhibition of Ca(2+) calmodulin dependent processes with trifluoperazine resulted in an increase in gamma-GTP activity. To study the role of a pkC-dependent pathway the effects of low doses of staurosporine were evaluated. Under these experimental conditions an increase in gamma-GTP activity was observed. It was then investigated whether these signal transduction pathways could interact with the FSH-stimulated cAMP dependent pathway to regulate gamma-GTP activity. Increase in extracellular Ca2+ concentration, the addition of 4Br-A23187 or the blockage of voltage-dependent Ca2+ channels did not modify FSH-stimulated gamma-GTP activity. However, staurosporine produced an additional increase in FSH-stimulated gamma-GTP activity and this effect was also observed when cells were stimulated with dbcAMP. In summary, our data are consistent with an inhibitory role of Ca(2+) calmodulin- and pkC-dependent pathways in the regulation of basal gamma-GTP activity. Similar to what has been shown for other Sertoli cell parameters, a pkC dependent pathway can interact with the FSH-stimulated cAMP-dependent pathway. The precise steps involved in this interaction are still unknown. PMID- 9401821 TI - Influence of oxygen tension on reactive oxygen species production and human sperm function. AB - Human spermatozoa were exposed to the reduced form of nicotinamide adenine dinucleotide phosphate (NADPH) to stimulate endogenous production of reactive oxygen species (ROS). They were incubated under a gas phase of 5% CO2/90% N2/5% O2, or 5% CO2/95% air (20% O2) to investigate whether a lower than atmospheric oxygen tension in the gas phase of the incubator limited the endogenous production of ROS by human spermatozoa and thus was able to reduce the cytotoxic effects of ROS on sperm function. Exposure of human spermatozoa or exogenous NADPH induced an 8-fold higher production of superoxide anion under ambient vs. low oxygen tension. This marked difference in the stimulation of superoxide anion generation was associated with significantly different sperm motility parameters, according to the oxygen tension in the gas phase of the incubator. Whereas under 5% oxygen the percentage of motile spermatozoa was unaffected by the presence of NADPH, all of the motility parameters recorded under an atmosphere of 5% CO2 in air were dramatically affected, not only compared to their respective controls, but also compared to the motility parameters observed under low oxygen tension. The presence of superoxide dismutase plus catalase protected spermatozoa against the toxic effects of NADPH, confirming a cause/effect relationship between the increased superoxide production and reduced sperm function. Even at a concentration of NADPH which did not alter the percentage of motile spermatozoa, hyperactivated motility and acrosome reaction were significantly lower under an atmosphere of 5% CO2 in air compared to a gas phase of 5% CO2/90% N2/5% O2. These results suggest that there is an advantage in using 5% O2 rather than 20% O2 in the gas phase of the incubator to prevent the excessive production of ROS by spermatozoa and related alterations of sperm functions. This may be of clinical value in fertilization programmes. PMID- 9401822 TI - Semen analysis performed by different laboratory teams: an intervariation study. AB - Some recent studies have indicated that sperm concentration has decreased during the last 50 years. However, comparisons between laboratories have revealed that geographical differences seem to exist and that any decrease may not be global. One point of criticism concerning comparison of results from different laboratories has been that some of the discrepancies detected could reflect the lack of standardized methods used in the different laboratories. Four teams, each consisting of one physician and one technician from groups which have recently published data on semen quality, met in order to evaluate the variability between their laboratories on semen analysis. Twenty-six fresh semen samples from unselected men were analysed. The groups analysed the samples according to the normal practice in their laboratories, using their own equipment. The variation between laboratories was estimated through a random effects model. For sperm concentration and semen volume assessment a remarkable consistency between laboratories was detected, in contrast to the very considerable inter-individual variation. For sperm motility and morphology assessments interlaboratory consistency was much poorer. In conclusion, evaluation of sperm motility and morphology characteristics requires further standardization in order to achieve comparable data from different laboratories. However, semen volume and sperm concentration are characteristics which can be compared reliably between laboratories, when similar methodologies are used. PMID- 9401823 TI - Sperm counts in semen of farm animals 1932-1995. AB - In some countries, sperm counts in normal human semen seem to have declined over the last 50 years. If this decline is real and due to environmental factors, falls might also be seen in sperm numbers in the semen of farm animals. Sperm counts are available for bull, boar and ram from the early 1930s, obtained using techniques similar to those used for human semen. Data have been obtained from the literature between 1932 and 1995 from 137 studies involving bulls, 76 involving boars and 130 involving rams. All were normal adult animals, from which semen was collected regularly but at a frequency which would not be likely to cause a fall in sperm counts. The references were obtained systematically from Animal Breeding Abstracts, and where possible the original articles were consulted to obtain mean values for each study; where the original reference was not easily obtainable, values were taken from the abstract. The bull data showed no correlation of sperm count with year of publication (r2 = 0.000), for the boars there was a slight but non-significant positive correlation (r2 = 0.041), and for the sheep there was a slight, but significant, rise in sperm counts with time (r2 = 0.124 for sperm counts and 0.126 for total sperm per ejaculate; not all authors gave both values). It would appear that, if the fall in human sperm counts is real, then it must be due to something which is not affecting farm animals. PMID- 9401824 TI - Estimating familial and genetic contributions to variability in human testicular function: a pilot twin study. AB - The biological basis for the remarkably wide variation in sperm output between and within men remains unclear. Although some contributing factors have been identified, the familial and genetic contributions to variation in human sperm output have been little studied, although such sources of variation are known to be significant in experimental animals. In order to identify such familial and genetic factors in a classical twin study design, we recruited monozygotic and dizygotic twins aiming to study the degree of resemblance between genetically identical and non-identical twins in sperm output, testis size and testicular endocrine function. From an approach to 160 twin pairs on the Australian Twin Register database, eventually 17 pairs of male twins (11 monozygotic and six dizygotic) participated in this study. Sperm concentration and output, right testis size and SHBG all exhibited a strong familial effect but a genetic component could not be confirmed. Total and free testosterone exhibited a genetic component. Although identifying for the first time a clear familial component to normal human spermatogenesis, this study had insufficient power to determine whether there was a genetic component, nor could its design distinguish between genetic or shared (early) environmental determinants for these familial effects. Implications for future twin studies of human testicular function including stronger recruitment strategies, multi-centre studies and surrogate variables for human spermatogenesis are suggested. PMID- 9401825 TI - Apoptosis as a mechanism of germ cell loss in elderly men. AB - In comparison with other mammals, human spermatogenesis is known to be an inefficient process, of which germ cell degeneration is a normal part. This study was performed to determine the mechanism of cell death in the testes of elderly men. Testes from 20 patients undergoing orchidectomy for prostate cancer were fixed, sectioned and processed for the detection of apoptosis using the in situ end-labelling technique. In addition, the formation of DNA ladders, a hallmark of apoptosis, was also investigated. Occurrence of apoptosis was not confined to a particular germ cell population but comprised all types of germ cells. Sertoli cell apoptosis was not encountered. The numbers of degenerating germ cells were determined per standard reference area, but no significant relationship was found between the mean values and age or testis weight. Analysis of the median values for germ cell death per reference area suggested that apoptosis occurs in clusters within the testis but is a rare occurrence outside these areas. It is concluded that spontaneous apoptosis can mediate germ cell death in a variety of cell types in the aged human testis. PMID- 9401826 TI - Histochemical demonstration of zinc ions in ejaculated human semen. AB - A revised in-vitro technique for autometallographic demonstration of chelatable zinc in the human ejaculate is presented, and the localization of the loosely bound pool of zinc ions is described in semen smears and at the ultrastructural level. In semen smears, black autometallographic (AMG) grains indicated the presence of zinc ions dispersed between the spermatozoa. These AMG grains have the same size as grains associated with the sperm tail and may have the same origin. EM analysis of AMG-developed smears fixed in osmium suggested that the detected zinc ions might be related to huge protein molecules present in semen and adhering to the surface of the spermatozoa. Spermatozoa in AMG-stained smears exhibited zinc ions in the midpiece and head, and also joined to the membrane of the tail. Washed spermatozoa exhibited zinc ions only within the midpiece. Ultrastructurally, they were found located in the helecine mitochondria. A few grains were found in the acrosome of the washed spermatozoa. Treatment with the chelating agent DEDTC resulted in complete bleaching of the zinc staining. These findings and the fact that calcium EDTA acid blocks the plasma and surface staining, but not the acrosomal and mitochondrial staining, suggest that chelatable zinc ions exist in two separate pools in human semen. PMID- 9401827 TI - Effect of tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN gamma) on human sperm motility, viability and motion parameters. AB - Male genital tract infections and non-specific inflammatory conditions may be associated with unexplained infertility. Previous studies have shown the presence of cytokines such as tumour necrosis factor-alpha (TNF-alpha) and interferon gamma (IFN-gamma) in the semen of infertile men. However, the mechanism of effect of these cytokines on human sperm function is still controversial. The present study was undertaken to investigate the in-vitro effects of TNF-alpha and IFN gamma on human sperm motion, viability and the hypoosmotic swelling test (HOST). Washed spermatozoa from normal volunteers (n = 9) were incubated in the presence/absence of TNF-alpha (1 microgram/mL) plus IFN-gamma (0.1 microgram/mL). Sperm motility, viability, HOST, and video sequences were recorded at different time intervals (0, 30, 60 and 180 min). Sperm motion parameters were analysed using computer-assisted semen analysis. There was a time-dependent negative effect of TNF-alpha plus IFN-gamma on sperm motility, viability, HOST, and lateral-head displacement (ALH). The maximum decrease was observed between 60 and 180 min for sperm motility (50.8 +/- 5.6%), viability (52.8 +/- 4.0%), HOST (38 +/- 2%) and ALH (4.7 +/- 0.1 microns) compared to control samples (62.2 +/- 2.8, 62.4 +/- 2.9, 58 +/- 4, and 5.3 +/- 0.4, respectively; All p < 0.05). There was no significant effect on sperm straight-line velocity and mean linearity when compared to control. These data suggest that the common inflammatory cytokines TNF-alpha plus IFN-gamma have only partial detrimental effects on sperm motility, viability, membrane integrity and lateral head displacement, which may contribute to the poor fertilizing potential of human spermatozoa during inflammatory conditions. PMID- 9401828 TI - Stage-specific degeneration of germ cells in the seminiferous tubules of non obese diabetic mice. AB - The cause of fertility problems in insulin-dependent diabetes is largely unknown. To evaluate the role of autoimmunity-associated phenomena in the testis as a possible cause of the derangement in spermatogenesis, the stage-specific apoptosis of germ cells in the insulitis phase of pre-diabetes was quantified in the testes of non-obese diabetic (NOD) mice. The seminiferous epithelium of normal BALB/c and NOD mice contained cells positive for in-situ end-labelling (ISEL) of DNA. ISEL-positive germ cells formed clusters in the seminiferous epithelium of the NOD mice in marked contrast to the seminiferous epithelium of the BALB/c mice, which contained only individual cells positive for ISEL. ISEL positive cells were present in the basal and luminal compartments of the epithelium. Ultrastructural analysis and demonstration of externalized phosphatidyl serine confirmed that the cells were undergoing apoptosis. The ultrastructurally apoptotic cells included spermatogonia, spermatocytes and spermatids. In cytological squash preparations of segments of seminiferous tubules from NOD mice aged 17-20 weeks, the number of ISEL-positive cells/mm tubule was significantly lower in segments at stages I-II of the seminiferous epithelial wave but higher at stages III-IV in comparison to BALB/c mice. The numbers of ISEL-positive cells/mm tubule in the other stages were similar in the two strains of mice. Analysis of 32P-3'-end labelled DNA from the testes showed that the BALB/c mice had relatively more DNA fragmentation than did the NOD mice. These data suggest that autoimmune insulitis in the NOD mice is associated with increased amounts and abnormal stage distribution of apoptosis in the seminiferous epithelium, resulting in derangement of spermatogenesis. PMID- 9401829 TI - The safety and tolerability of single intravenous doses of lubeluzole (Prosynap) in healthy volunteers. AB - The safety and tolerability of single escalating doses of lubeluzole were evaluated in healthy male volunteers in 2 studies. In the first of 2 randomized, single-blind, placebo-controlled, dose-escalation studies, 6 subjects received single 30-minute infusions of 2.5, 5, and 10 mg of lubeluzole, and 2 additional subjects received placebo. In the second study 6 different subjects received a 1 hour infusion of 15 mg of lubeluzole, 5 of whom received the 20-mg dose, and 2 received 25 mg of lubeluzole. Two additional subjects received placebo. Small increases and decreases in PQ, QRS, QT, QTc, and QTm intervals were noted after infusion of all lubeluzole doses and placebo, however, these changes were within the normal ranges for these values except for the QTc for the 25-mg dose of lubeluzole. Significant prolongation of the QTc interval was observed at the end of the 1-hour infusion in both subjects receiving the 25-mg dose of lubeluzole. No clinically relevant changes in systolic time intervals, heart rate, blood pressure, and clinical laboratory values were noted in subjects receiving 2.5-25 mg of lubeluzole or placebo. Adverse experiences, predominantly lightheadedness and dizziness, were reported by subjects receiving doses of lubeluzole greater than or equal to 10 mg. Lubeluzole, administered as single intravenous doses of 2.5-15 mg, is safe and well tolerated in healthy male volunteers. PMID- 9401830 TI - A single and multiple dose bioavailability study with carbamazepine 400 mg retard tablets with reference to enzyme autoinduction and circadian time differences. AB - Both single and multiple dose bioequivalence studies are required to assess the quality of modified release formulations of drugs. In bioequivalence studies of drugs with enzyme autoinducing properties such as carbamazepine (CBZ), the standard multiple dose study design must be modified to guarantee equivalence of drug elimination. This problem was considered with regard to carbamazepine 400 retard AWD (test) whose bioavailability relative to a listed reference (Tegretal 400 retard) was studied in 2 randomized, open, crossover studies both with 18 healthy volunteers of Caucasian origin (20-36 years, 61.5-92 kg, 172-195 cm). The single dose study was done with 400 mg CBZ. Serum concentration time profiles of CBZ and its active metabolite CBZ-10,11-epoxide were determined until 144 h after administration. The multiple dose study was performed with 400 mg CBZ b.i.d. for 15 days (first 2 days: 200 mg b.i.d.) followed by a 7-day study with the alternative investigational product. 24-hour serum concentration time profiles of CBZ and its metabolite were measured on days 15 and 22 of the study. The quantitative drug analysis was done with an HPLC method the quality of which fulfilled the requirements of bioequivalence studies. Test was considered bioequivalent to reference with regard to the extent of absorption, if the 90% confidence intervals of the AUC0-infinity ratio (single dose) and AUC0-24h ratio (multiple dose) were within the range of 0.80-1.25, and with regard to rate of absorption if the 90% confidence intervals of the Cmax/AUC ratio (single dose) or AUCF0-24h ratio were within 0.70-1.43. The point estimators (90% confidence limits) of the AUC ratio (test/reference) of CBZ were 0.979 (0.94, 1.02) for the single and 1.01 (0.947, 1.076) for the multiple dose comparison. The point estimator (90% confidence limits) of the Cmax/AUC ratio was 0.989 (0.959, 1.020) and of the AUCF0-24h ratio 1.066 (0.937, 1.212). There were no circadian time differences in any pharmacokinetic parameter. IN CONCLUSION: Carbamazepine 400 retard AWD tablets were bioequivalent to reference with regard to extent and rate of absorption after both single and multiple dose administration. PMID- 9401831 TI - An alternative analysis for crossover studies that accounts for between-group disparities in drug response. AB - BACKGROUND: In crossover clinical trials comparing completely different treatments patients tend to fall apart into different populations: those who respond better to treatment 1 and those who do so to treatment 2. The correlation between treatment response in such trials is negative. The current ANCOVA analysis for crossover studies does not allow for correlations being negative, and is, therefore, not adequate to test this kind of trial. OBJECTIVE OF STUDY: To study whether matrix algebra provides a more appropriate approach for this purpose. RESULTS AND CONCLUSIONS: Using a mathematical model as well as hypothesized examples I demonstrate that matrix algebra of 2 pairs of cells of the same order not only allows for negative correlations in a crossover design but also provides enough power to test both treatment and carryover effect. PMID- 9401832 TI - A reliable and convenient parameter of the rate of paracetamol absorption to measure gastric emptying rate of liquids. AB - The rate of paracetamol absorption represents gastric emptying rate (GER) of liquids. Thus, the liquid GER is assessed by conventional pharmacokinetic parameters such as the maximum concentration and the time to maximum concentration after oral administration of paracetamol. However, the conventional parameters are subject not only to the rate but also to the extent of absorption. For the reliable assessment of GER we have proposed a new parameter, the C0.5/C0.25 ratio, for the rate of paracetamol absorption without being affected by the extent of absorption. Of 15 healthy male volunteers, 9 orally received 10 mg/kg of paracetamol with 200 ml of water as the "normal" GER group, and the other 6 took 10 mg/kg of paracetamol with 200 ml of a liquid nutrient (200 kcal/200 ml), which delays GER, as the "delayed" GER group. Blood samples were obtained at t = 0 (pre-dose), 0.25, 0.5, 0.75, 1.0, 1.5, 2.0, 3.0, 4.0, 6.0, and 8.0 hours (post-dose). In each subject, GER was assessed by the conventional parameters, the C0.5/C0.25 ratio, and the Wagner-Nelson method which provides an accurate estimate of the drug absorption rate. Using the C0.5/C0.25 ratio and the Wagner-Nelson method we could more clearly differentiate the delayed GER group from the normal GER group than by using the conventional parameters. This suggests that the C0.5/C0.25 ratio and the Wagner-Nelson method may be more reliable than the conventional parameters in detecting a delay in GER. Further, it should be noted that the C0.5/C0.25 ratio can be calculated from only 2 blood samples while the Wagner-Nelson method requires repeated blood sampling. PMID- 9401833 TI - Crossover comparison of the pharmacokinetics of amlodipine and felodipine ER in hypertensive patients. AB - The pharmacokinetics of amlodipine 5 mg and felodipine ER (extended release) 5 mg o.d. after single and 2 weeks of repeated oral doses, were compared in 28 essential hypertensive patients using a crossover design. As a secondary parameter the effects of the drugs on blood pressure were assessed. Significant differences were found between all principal pharmacokinetic variables, when comparing the 2 treatments after both single and repeated dosing. The coefficients of variation of maximal drug concentration and AUC after single dosing and at steady-state were significantly higher for felodipine ER than for amlodipine. After repeated dosing the peak-to-trough plasma concentration ratio were 1.58 and 4.43 (p < 0.001) for amlodipine and felodipine ER, respectively. Both drugs lowered systolic and diastolic blood pressure to the same extent after 2 weeks of repeated dosing. No significant differences between the blood pressure lowering vs time profile of the 2 drugs were encountered. In conclusion, the interpatient drug concentration variability and the peak-to-trough plasma concentration ratio were more favorable for amlodipine compared to felodipine ER. It remains to be established whether these characteristics are also reflected in a more smooth and consistent blood pressure control. PMID- 9401834 TI - Metoclopramide plasma concentration in neonates. AB - During the course of a multiple-dose metoclopramide (M) oral treatment, M plasma concentrations were measured just before (Cmin) and 1 hour after the administration (C1h) at steady-state in 5 (3 premature and 2 term) neonates. Mean Cmin was equal to 91.6 +/- 45.5 ng/ml and higher than C1h (87.4 +/- 43.2 ng/ml), but not significantly. A significant negative correlation was found between Cmin plasma concentration and gestational age as well as with postconceptional age, suggesting that the lower the gestational and postconceptional age, the lower the metoclopramide dosage should be. PMID- 9401835 TI - No need to adjust the dose of 311C90 (zolmitriptan), a novel anti-migraine treatment, in patients with renal failure not requiring dialysis. AB - 311C90 ("Zomig", zolmitriptan), is a novel and selective, centrally and peripherally acting 5 HT1B/1D receptor agonist in development for the acute, oral treatment of migraine. We have conducted a parallel group study in patients with moderate/severe renal impairment (creatinine clearance < or = 40 ml/min) and age- and sex-matched healthy volunteers (creatinine clearance > or = 60 ml/min). All subjects received a single, 10 mg dose of 311C90. Mean peak concentrations of 311C90 and its pharmacologically active N-desmethyl metabolite (183C91) were similar in both groups although AUC0-infinity for 183C91 was increased by 35% in the renally impaired patients. Other pharmacokinetic parameters were little changed apart from the expected reduction in CLR and urinary recovery and a small increase of 0.9 and 1.0 h, respectively, in the mean half-lives of 311C90 and 183C91. For the 2 inactive metabolites, the N-oxide (1652W92) and the indolacetic acid (2161W92), mean peak concentrations were approximately 3 times higher in renally impaired patients than in healthy volunteers and AUC0-infinity was 6-7.5 times higher. CLR for these metabolites was approximately 90% lower in renal impairment and half-life of both was increased approximately 3-fold. Baseline blood pressures were higher in the renally impaired group. After 311C90 there was a transient, small increase in blood pressure in both groups. There was little difference in the increase in diastolic blood pressure between the groups (16 mmHg in both) but the rise in systolic blood pressure was greater in the renally impaired group (23 mmHg vs 16 mmHg in healthy subjects). The lack of substantial changes in the plasma concentrations of both parent compound and 183C91, and the similarity of the changes in blood pressure, in renally impaired subjects compared to healthy volunteers suggest that there is no reason to adjust the dose of 311C90 in patients with renal impairment. PMID- 9401836 TI - A new route, jet injection for anesthetic induction in children--III. Ketamine pharmacokinetic studies. AB - The jet injector route for ketamine was used on 30 children 1-6 years of age undergoing various surgical procedures. A randomly selected dose of 2.5, 3.5, or 6.0 mg/kg of ketamine was given to induce anesthesia. Peak plasma ketamine levels did not follow a simple arithmetic increment related to dose. Dosage based on mg/m2 body surface area or mg/kg body weight provided similar blood levels of ketamine. The beta-phase t1/2 of ketamine in these children was shorter than that found in adults. Considerable individual variability was observed in both the plasma levels to a given dose of jet-injected ketamine and in the beta-phase t1/2. The ketamine beta-t1/2s were not dose related. PMID- 9401837 TI - Influence of acute renal failure on FPIA rapid serum assay of midazolam and its main metabolite. AB - We recently developed a simple and fast assay technique, providing the possibility of monitoring of midazolam (M) during sedation. We compared HPLC vs FPIA for the measurement of the sum M plus alpha 1-hydroxymidazolam (OM), its main and pharmacologically active metabolite, in the serum of sedated ICU patients; this activity referred to as M-like. We identified certain patients in whom M-like activity appeared abnormally high in comparison with HPLC assays. Their common denominators were: long-term sedation with M, and seriously impaired renal function. Further, the conjugates of OM (OMG) accumulated in patients with acute renal failure could contribute to the sedation. We compared the metabolic and analytic behavior of M, OM, and OMG in 2 groups of sedated patients either presenting with normal renal functions (group 1) or with a picture of acute renal failure (group 2). Blood samples were assayed by HPLC and by FPIA and analysis was performed before and after hydrolysis of OMG. Before hydrolysis there was a dramatic accumulation of OMG in the patients of group 2, HPLC vs FPIA results were not different within group 1, while in group 2 the FPIA response exceeded that of HPLC. After hydrolysis, measurement by HPLC was greatly increased in group 2, in each group (vs HPLC) and from one group to another, the FPIA signal (the M-like activity) showed a significant increase. It would be important to take OMG into account as a coprotagonist in sedation whenever circumstances predispose to its accumulation. PMID- 9401838 TI - Straight ileoanal anastomosis with multiple ileal myotomies as an alternative to pelvic pouch. AB - An alternative technique of restorative proctocolectomy, by means of straight ileoanal anastomosis with multiple myotomies (SIAM) of the terminal ileum in 15 patients, nine with familial adenomatous polyposis (FAP) and six with ulcerative colitis (UC) is reported. SURGICAL TECHNIQUE: Eight to ten longitudinal myotomies (3-4 cm long, on three different circumferential sites) were performed on the terminal ileum for a total length of 12-14 cm. CLINICAL RESULTS: At a mean follow up of 44 months (range 3-84 months) from the closure of the ileostomy, daytime continence was achieved in all the patients; stool frequency per 24 hours (+/- SD) was 4.1 +/- 1.8 for FAP patients and 5.8 +/- 1.7 for UC patients; nocturnal defecation was 1.0 +/- 0.5 and 1.2 +/- 0.8 for FAP and UC patients respectively; frequent nocturnal soiling was present in 2/5 of UC patients, and in 3/9 of FAP patients. SIAM failed in one UC patient that was converted to an ileoanal reservoir because of poor functional result. Signs of ileal mucosal inflammation were never observed at endoscopic examination. Histopathological assessment showed no evidence of acute terminal ileitis. MANOMETRIC FINDINGS: A significant postoperative reduction in anal resting pressure was observed after SIAM. Neither the absence of anal inhibitory reflex nor the presence of high pressure waves generated in the terminal ileum during air insufflation were related to the presence of soiling. The closure of the loop ileostomy was followed by an increased capacity and distensibility of the terminal ileum. Values of neorectal compliance were similar in FAP and UC patients although FAP patients were able to reach higher values of maximum tolerated volume and pressure. CONCLUSIONS: 1) SIAM can be an alternative to pelvic pouch in patients who have undergone restorative proctocolectomy when the construction of the pouch is not feasible. 2) The functional result observed after SIAM has been shown to be similar to that observed after pouch construction. PMID- 9401839 TI - Risk of invasive carcinoma in colorectal adenomas assessed by size and site. AB - BACKGROUND: The risk of invasive carcinoma developing in colorectal adenomas is influenced by a number of characteristics of both patients and adenomas, and the composition of the sample analysed. PATIENTS AND METHODS: Between 1978 and 1993 more than 20,000 polyps were prospectively documented at the Erlangen Registry of Colorectal Polyps, and analysed statistically by logistic regression. RESULTS: The size of the adenomas proved to be the most important factor for adenomas equal to or larger than 15 mm as compared with smaller lesions. In 5,137 diminutive adenomas (< or = 5 mm) invasive carcinoma was never found. Adenomas in the right-sided colon had a lower risk than those in the left colon or rectum, but with increasing adenoma size, the malignancy rate showed a right-sided shift. In adenomas of up to 36 mm in diameter, invasive carcinoma was found more often when they were located in the rectum or left colon while adenomas larger than 36 mm were more likely to harbour invasive carcinoma when located in the right or left colon rather than in the rectum. CONCLUSIONS: A multivariate analysis of 11,380 adenomas detected at the first total colonoscopy showed that the factors size and site, both of which can be assessed by endoscopic inspection alone, were found to enable a statistically and clinically adequate assessment of the malignancy risk. PMID- 9401840 TI - Extended mesenteric excision in right hemicolectomy for carcinoma of the colon. AB - Between 1979 and 1989, 169 patients had a curative operation for right sided colonic cancer. A retrospective analysis of the incidence and degree of lymph node metastasis was performed in all and survival rate was determined in 144 patients who could be followed over a period of 5 years or more. In all patients, dissection involved the removal of right colon (i.e., caecum, ascending colon, and right side of transverse colon). Dissection of regional lymph nodes in 84 patients (group 1) involved the removal of mesocolic lymph nodes related to the segment of the removed intestine. In 60 patients (group 2) dissection was extended to the nodes situated anterior to mesenteric and retropancreatic vessels. Morbidity and mortality rates were similar in the two procedures. The number of lymph nodes and the level of apical node examined were significantly different in the two groups. The 5-year survival rates showed no statistically significant difference, but in group 2 three of the nine patients with metastasis to N4 nodes are free of disease, surviving at 7, 12 and 14 years, respectively. The principle of extensive lymph node dissection is proposed as a procedure that supplies more accurate staging and might reduce the incidence of loco-regional recurrence. PMID- 9401841 TI - Magnetic resonance imaging in the management of fistula in ano. AB - Fistula-in-ano is a common condition in which accurate diagnosis of the fistula track is essential as inadequate assessment and surgical treatment may lead to multiple unnecessary operations and may also render the patient incontinent. Several studies have suggested that Magnetic Resonance Imaging (MRI) can accurately identify the fistula track in relation to the sphincter complex. The aim of this study was to investigate the value of the routine use of completely non-invasive pre-operative MRI in patients with suspected fistula-in-ano. Each scan was reported by a consultant radiologist on two occasions to determine whether the radiologist's opinion had changed and/or become more accurate with further experience. Surgical assessment of the fistula was performed under general anaesthesia by one surgeon without knowledge of the result of the MRI scan. The results of the surgical assessment and the MRI scan were compared and the surgical procedure completed. Thirty three patients with a clinical diagnosis of fistula-in-ano were treated and 27 subsequently confirmed to have a fistula. MRI detected 42% of tracks, identified correctly on initial assessment which increased to 50% at the end of the study, 63% and 74% of internal openings, 33% and 46% of external openings and 50% and 33% of abscesses. These data suggest that there is a learning curve for radiologists undertaking MRI scanning for fistula in ano, this is probably because the pathology of fistula in ano and anatomy of the anal sphincter complex are relatively new to radiologists. Routine MRI scanning of patients with fistula-in-ano is not necessary but there may be a role for MRI in assessing complex or difficult fistulae. PMID- 9401842 TI - Reliability of pudendal nerve terminal motor latency. AB - AIM: To evaluate reliability of Pudendal Nerve Terminal Motor Latency (PNTML). METHODS: Forty healthy subjects, 24 women and 16 men, and eight female patients were included. Four patients had idiopathic faecal incontinence and 4 an anal sphincter rupture after child-birth. PNTML measurement was performed by two observers with the patient in left lateral and supine position. Examinations were repeated on another day to evaluate intraindividual reproducibility. RESULTS: Interobserver reproducibility was 92%-116% for PNTML. Degree of agreement for PNTML between left lateral and supine position was 86%-111%. Intra-individual reproducibility in the supine and left lateral positions was 89%-109% and 88% 113% respectively. Normal values for mean PNTML were higher in women compared with men, 1.91 msec (2 SD, 0.52 msec) and 1.74 msec (2 SD, 0.33 msec) respectively, t = 2.44, 37 DF, P < 0.01. CONCLUSIONS: Reliability of PNTML in terms of interobserver and intraindividual reproducibility was high. Women had higher normal values for PNTML than men. PMID- 9401843 TI - Emergency large bowel surgery: a 15-year audit. AB - OBJECT: To evaluate the management of patients presenting with colorectal emergencies. METHOD: Computerized audit of patients undergoing urgent/semi-urgent surgery in the Colorectal Service, University Department of Surgery, Wellington School of Medicine, NZ. RESULTS: 246 patients underwent major emergency or semi emergency operations. Consultants performed 144 operations. The complications of cancer and diverticular disease were the commonest indications for surgery. Patients with inflammatory processes required significant perioperative nutrition. The disease site varied with the pathology. Overall the sigmoid colon was the commonest. Resection and anastomosis was generally performed for right sided lesions whereas Hartmann's operation was the commonest procedure for more distally situated non neoplastic lesions. A loop diverting stoma was used most commonly in patients with obstructing cancer. The most frequent post-operative complication was urinary tract infection. Four patients developed pulmonary embolism, 2 ARDS, 4 myocardial infarction and 1 CVA. Persistent intra-abdominal sepsis requiring drainage occurred in five patients. There were 6 anastomotic leaks. 3 patients were re-operated upon to relieve post-operative small bowel obstruction. The overall post-operative mortality rate was 6.9%. CONCLUSION: A cautious policy of resecting right sided lesions and either diversion or resection without anastomosis for patients presenting acutely with left-sided colonic lesions resulted in a low overall mortality rate. PMID- 9401845 TI - Ambulatory anorectal manometric findings in patients before and after haemorrhoidectom. AB - The physiological abnormalities in piles before and after surgery were studied by an ambulatory prolonged anorectal manometric technique. Eighteen consecutive patients (12 men, 6 women; mean age 43.6 [standard error of mean, 3.3] years) with 3 prolapsed irreducible piles were prospectively recruited. Haemorrhoidectomy was performed with excision of 3 piles. The anal and rectal pressures were monitored before and at a mean 7.8 (1.5) weeks after surgery when the wounds had healed, for a mean period of 361.3 (47.8) min. The maximum anal pressures dropped significantly from a mean 325 (15.5) mmHg before to 213 (24.9) mmHg after surgery (P < 0.05). Ultraslow wave activity was clearly identified in 11 of 18 patients (61%) before surgery, but not found on post operative studies. The maximum rectal pressures were also significantly reduced after surgery (196.8 [23.2] mmHg before, 75.5 [10.6] mmHg after; P < 0.05). These findings help to confirm that haemorrhoidectomy leads to changes in the anorectal physiological findings. PMID- 9401844 TI - Screening for colorectal neoplasia with faecal occult blood testing compared with flexible sigmoidoscopy directly in a 55-56 years' old population. AB - Reduced mortality from colorectal cancer may be achieved by screening with faecal occult blood testing. Screening for neoplasia in the rectum and sigmoid colon with flexible sigmoidoscopy is suggested to be more effective, particular among persons between 50 and 60 years of age. A cohort of 6367 persons 55-56 years of age were randomised to screening with rehydrated Hemoccult II tests (HII group) or with flexible videosigmoidoscopy directly (FS group). In the HII group 59% (1893/3183) attended, compared to 49% (1353/3184) in the FS group. Of the 1893 persons who attended in the HII group, 4% had a positive HII test and in 13% (10/78) of them a neoplasm > or = 1 cm in the rectum or sigmoid colon was diagnosed by endoscopy. The corresponding rate in the FS group was 2.3%. Overall the number of persons with a neoplasm > or = 1 cm diagnosed in the HII group was 10 and in the FS group 31. A subgroup in the flexible sigmoidoscopy group, who also performed rehydrated HII tests, showed a sensitivity of the HII test for neoplasia > or = 1 cm of 26% and a specificity of 95.6%. To find a neoplasm > or = 1 cm in the rectum or sigmoid colon, 44 examinations were needed when using flexible sigmoidoscopy directly and 7 examinations when only those with positive HII tests were examined. In mass screening for neoplasia in the rectum and sigmoid colon, the relatively low prevalence of colorectal neoplasia at 55-56 years of age makes primary selection with rehydrated Hemoccult testing an alternative to the resource-consuming endoscopy of all invited persons. PMID- 9401846 TI - Is mechanical bowel preparation really necessary for elective left sided colon and rectal surgery? AB - The value of mechanical bowel preparation for elective left sided colorectal surgery is debatable. This retrospective study evaluates the incidence of wound infection, wound dehiscence, abdominal/pelvic collections and anastomotic dehiscence between patients who received mechanical bowel preparation [MBP] (n = 61) and those who did not (n = 75). The case notes of 136 consecutive patients undergoing elective left sided colorectal surgery over a three year period in a district general hospital were reviewed. The incidence of infective and anastomotic complications between the two groups was not significantly different. There were two post-operative deaths, both in patients receiving MBP. We therefore conclude that the role of MBP in the era of systemic antibiotics must be questioned. A prospective randomised multicenter trial recruiting an adequate number of patients undergoing elective left sided colorectal procedures would clarify this long standing debate. Note: Presented at the Spring Meeting of the Minnesota Surgical Society in Rochester, Minnesota, USA, May, 1996 and to the American Society of Colon and Rectal Surgeons (ASCRS), Seattle, Washington, June 1996, and published in abstract form in Diseases of the Colon and Rectum (1996) 39:A47. PMID- 9401847 TI - Correlation of pudendal nerve terminal motor latency with the results of anal manometry. AB - Denervation to the external anal sphincter is commonly found in disordered defaecation. AIM: To determine whether a correlation exists between pudendal nerve terminal motor latency (PNTML) and anal manometry and what influence an external sphincter defect (ESD) has on any correlation. METHOD: Sixty seven consecutive patients (23 constipated, 44 incontinent) were analysed. All had results available for PNTML and anal manometry. Anal ultrasound performed in the later part of the study period was available in 46 patients. RESULTS: A significant negative correlation was found between the mean PNTML and squeeze pressures (SP) for incontinent patients (r = -0.32, P = 0.037). No significant correlation was seen in constipated patients. A coexisting ESD was found in 57% of the 46 patients studied. In those without an ESD a significant negative correlation was found between mean PNTML and SP (r = -0.50; P = 0.026). No correlation was found in patients with an ESD. Age did not significantly affect the PNTML or SP results, but was associated with a reduced resting pressure (r = 0.34; P = 0.005). CONCLUSIONS: The PNTML was significantly correlated with SP in patients with incontinence and in the subgroup of patients without an ESD. In the assessment of disordered defaecation PNTML is therefore recommended as an adjunct to anal ultrasound. PMID- 9401848 TI - Electrostimulated gracilis neosphincter for faecal incontinence and in total anorectal reconstruction: still an experimental procedure? AB - The possibility of converting an easily fatiguable muscle like the gracilis muscle into a fatigue-resistant one using chronic electrostimulation has renewed interest in Pickrell's procedure. Between July 1991 and June 1996, 9 patients (2 M; 7 F) mean age = 45 y (range 14-72) underwent dynamic graciloplasty using Medtronic electrostimulators. Five patients had faecal incontinence (2 congenitally anomaly, 1 neurological, 2 post-operative) and 4 had a perineal colostomy performed either simultaneously (two cases) or at 3 to 4 years after abdominoperineal excision of the rectum. Early post-operative complications included distal tendon necrosis [1], perineal colostomy breakdown [1], detachment of the gracilis tendon [2] and seroma in the thigh [1]. Long-term complications included rectocele with faecal impaction in one patient with imperforate anus, anal stricture in one patient who had refashioning of a perineal colostomy, and displacement of the lead from the main nerve in 3 with external expulsion in 2. The patient with anal stricture was successfully treated with anoplasty but subsequently returned to an abdominal colostomy due to stricture recurrence 2 years later. The rectocele was successfully treated using a transvaginal approach. Electrical conversion of the muscle was completed in all patients but long term functional results are available for only 5 cases. Manometry revealed a significant improvement in anal pressure under electro-stimulation and the continence grading scale score significantly improved in 4 patients. The technique is applicable to a very selected group of patients with no other options but is still in the experimental phase and should not be performed outside controlled trials. Repeated hospitalisation and reoperations are often required although the complication rate may diminish and improve with experience. PMID- 9401849 TI - Solitary rectal ulcer syndrome--an underdiagnosed condition. PMID- 9401850 TI - "Rectal hypersensitivity in the irritable bowel syndrome". PMID- 9401851 TI - Hello out there! Results of J.A.H. readership survey. PMID- 9401852 TI - Response to "Out of wedlock births: what do the statistics really mean?". PMID- 9401853 TI - Assaultive violence in the community: psychological responses of adolescent victims and their parents. AB - PURPOSE: This article presents an overview of psychological responses to injuries due to assaultive violence, particularly gunshot wounds, in adolescents and parents. METHODS: Reviews were conducted of the psychological, medical, and public health literature to identify studies of violent injury in adolescent populations. Studies reviewed in this article include those reporting levels of gunshot and other violent injuries among pediatric patients in urban hospitals and those examining psychological responses of children and parents to violent injury and parental responses to potential death of a child. RESULTS: Existing literature indicates that the number of adolescent victims of assaultive violence is increasing. Adolescent victims of assaultive violence and their parents may experience significant psychological distress including school difficulties, guilt and fears of subsequent injury, and changes in risk behaviors and perceptions of life. CONCLUSION: Greater awareness of the psychological repercussions of injury due to assaultive violence for adolescents and their families is needed among medical professionals. Hospital based interventions, which may include psychological support for adolescents and their parents and referrals for academic evaluation and assistance, should be developed for adolescent trauma victims and their families. PMID- 9401854 TI - National cancer clinical trials: children have equal access; adolescents do not. AB - PURPOSE: To determine whether adolescents with cancer, who in comparison to younger patients have a higher cancer incidence and lower mortality reduction, have equal access to national cancer clinical trials. METHODS: The ethnic/racial distribution of 29,859 subjects < 20 years of age entered onto National Cancer Institute-sponsored clinical trials between January 1, 1991, and June 30, 1994, was compared with the expected distribution of patients of the same age in the United States. RESULTS: The Children's Cancer Group and Pediatric Oncology Group had 29,134 (97.6%) of the total study entries among < 20-year-old subjects during the 3.5 years of surveillance. The adult cooperative groups accounted for < 3% of the clinical trials entries in the 15-19-year age range. When analyzed nationally by region, the under-representation of the older adolescent subjects was universal. From other analyses, the two pediatric cooperative groups were estimated to have registered > 94% of the children < 15 years of age who were expected to have been diagnosed to have cancer, but only 21% of the cancer patients in the 15-19-year age group. CONCLUSIONS: The national pediatric cancer cooperative groups allow the majority of American children < 15 years of age and their families equal opportunity to access clinical cancer trials, regardless of race or ethnicity. Among patients 15-19 years of age, however, > 75% are not being enrolled by any cooperative group sponsored by the National Cancer Institute. Thus, older adolescents are disadvantaged with respect to access to the national clinical trials, regardless of their race or ethnicity. PMID- 9401855 TI - Adolescent under enrollment in national clinical research: it is time to ask why. PMID- 9401856 TI - Obtaining written parent permission for school-based health surveys of urban young adolescents. AB - PURPOSE: To document the process and implications of obtaining written parental consent for school-based health surveys of young adolescents. METHODS: As part of the evaluation of the Reach for Health prevention program, written parental permission was obtained for student participation in school-based health surveys conducted for three cohorts of seventh graders (N = 3253) enrolled in three urban schools serving predominately economically disadvantaged minority adolescents. Students in general, bilingual, and special education classes were eligible to participate. Rates were recorded for the number of forms returned by parents, parental consents and refusals, student consents and refusals, and surveys completed. Procedures for achieving acceptable rates of written parental permission and survey completion included daily communication between research and school staff during the consent form collection period, student and teacher incentives, provision of alternate activities for students without consent, and scheduling of multiple makeup surveys for absentee students. RESULTS: Survey completion rates met or exceeded preset goals and ranged from a low of 70% for Cohort A to a high of 83% for Cohort C. At least 89% of the parents in each cohort returned forms. Of forms returned, parent refusals ranged from a high of 18% (Cohort A) to a low of 12% (Cohort C). CONCLUSIONS: Obtaining written permission from parents for young adolescents to participate in school-based health surveys is possible in urban settings and has potential benefits in terms of community awareness and involvement in research and evaluation studies. It does, however, require a substantial commitment of program resources as well as significant planning and data collection prior to actual survey administration. PMID- 9401857 TI - Human subjects protection and parental permission in adolescent health research. PMID- 9401858 TI - Adolescents report their need for and use of health care services. AB - PURPOSE: The goals of this study were to describe student access to health care services, identify populations of students who remained in need of health care services, and highlight particular unmet needs for health care identified by these adolescents. METHODS: Students in Grades 9-12 attending 50 schools in Oregon completed the Youth Risk Behavior Surveillance Survey (YRBS). Questions requesting adolescents to report their need for specific types of health care, and access to general and specific types of care were added to the core YRBS. Multivariate logistic regression analysis was used to determine independent relationships between student or community characteristics and health care access or unmet needs for care. RESULTS: Almost 14,000 adolescents completed surveys, of whom 75% reported visiting one or more health care provider within the last 12 months. Nineteen percent of adolescents reported that they had not received 1 or more of 10 specific types of care when needed in the last year. Females, some racial/ethnic minorities, rural, and sexually active adolescents were more likely to report unmet needs for health care. Most frequently, adolescents reported they needed but did not receive care for an illness (7%) or for personal or emotional problems (6%). In addition, about 400 (3%) students reported they needed birth control that they did not receive. CONCLUSIONS: A majority of high school-age adolescents had visited health care providers within the year prior to study. However, the number of adolescents who reported unmet specific health care needs within the same time period remained substantial. PMID- 9401859 TI - Protecting against hopelessness and suicidality in sexually abused American Indian adolescents. AB - PURPOSE: The purpose of this study was to identify factors protective against the adverse health correlates of sexual abuse in reservation-based American Indian and Alaskan Native adolescents. METHODS: Data were taken from the National American Indian Adolescent Health Survey administered in 1988-1990 to 13,923 youths. Included in this analysis were 991 females and 166 males who reported a history of sexual abuse. Chi-square analysis was used to identify significant protective factors in sexually abused youths who did not report suicidality or hopelessness. Discriminant function analysis was used to determine which factors distinguished this group from those who experienced adverse health correlates. RESULTS: Separate multivariate analyses for boys and girls demonstrated that for girls, family attention, positive feelings toward school, parental expectations, and caring exhibited by family, adults, and tribal leaders were associated with absence of suicidality and hopelessness. For suicidality in boys, significant protective factors were enjoyment of school, involvement in traditional activities, strong academic performance, and caring exhibited by family, adults, school people, and tribal leaders. No significant protective factors against hopelessness were identified for boys. CONCLUSIONS: To minimize hopelessness and suicidal involvement among youth who have been sexually abused, strategies should be planned, implemented, and evaluated that support family caring and connectedness, strengthen school attachment and performance, and improve tribal connectedness. PMID- 9401860 TI - Confidential health care for adolescents: position paper of the Society for Adolescent Medicine. PMID- 9401861 TI - Driver education: position paper of the Society for Adolescent Medicine. PMID- 9401862 TI - Controversies in assisted reproduction and genetics. Does "EPF" have an identity? PMID- 9401863 TI - An update on the identity of early pregnancy factor and its role in early pregnancy. PMID- 9401864 TI - Early pregnancy factor: an unresolved molecule. PMID- 9401865 TI - An update on the identity of early pregnancy factor and its role in early pregnancy. PMID- 9401866 TI - Ethical principles of the Commonwealth Medical Association. PMID- 9401867 TI - In vitro fertilization with low-dose clomiphene citrate stimulation in women who respond poorly to superovulation. AB - PURPOSE: Our experience with IVF using low-dose clomiphene citrate for stimulation in "non-" and "poor" responders was reviewed and the treatment outcomes with the previous controlled ovarian stimulation cycles in which hMG and GnRH agonist were used were compared. METHODS: The treatment outcome in 11 non- and 20 poor responders having 30 and 53 clomiphene citrate IVF treatment cycles, respectively, were compared with the treatment outcome in the previous long protocol buserelin/hMG cycles. RESULTS: The clinical pregnancy rates per oocyte collection achieved in the first clomiphene citrate cycle in non (9.1%)- and poor (10%) responders were comparable to those achieved by poor responders (11.9%) who had buserelin/hMG using the long protocol. Although the numbers were small, a similar pregnancy rate could still be achieved in poor responders up to the third attempt using clomiphene citrate. CONCLUSIONS: IVF using long-protocol buserelin/hMG is more successful than using clomiphene citrate stimulation. However, this advantage may not be significant in those women with a previous poor response to buserelin/hMG. It is suggested that for such poor responders, three attempts of IVF in a clomiphene citrate cycle may offer a viable therapeutic alternative before reverting to more stressful, expensive, and time consuming treatment. PMID- 9401868 TI - The presence of hydrosalpinx may not adversely affect the implantation and pregnancy rates in in vitro fertilization treatment. AB - PURPOSE: To evaluate the effects of hydrosalpinx on the outcome of in vitro fertilization (IVF) treatment, a retrospective study was undertaken at a tertiary referral center for infertility. METHODS: Results of the first IVF treatment cycles in 144 patients from 1 January 1993 to 31 December 1995, who had tubal infertility only and were less than 38 years old, were reviewed. The duration/dosage of hMG used, serum estradiol level on the day of hCG, number of oocytes aspirated and fertilized, number of embryos replaced, implantation rate, clinical pregnancy rate, and pregnancy outcome were compared in patients with and without hydrosalpinx. RESULTS: The mean implantation rate and clinical pregnancy rate were similar in patients with or without hydrosalpinx. Both groups had similar ovarian responses and fertilization rates. There was no increase in clinical abortion in the hydrosalpinx group but ectopic pregnancies were more common in patients with hydrosalpinx. CONCLUSIONS: The presence of hydrosalpinx did not adversely affect the implantation and pregnancy rates in in vitro fertilization treatment when the results of the first cycle were compared. However, it can lead to a higher incidence of ectopic pregnancies. PMID- 9401869 TI - Outcome of IVF in DES-exposed daughters: experience in the 90s. AB - PURPOSE: The outcome of in vitro fertilization (IVF) in a group of infertile women with a history of in utero exposure to diethylstilbestrol (DES) was analyzed. Records from an academic IVF program were retrospectively reviewed. METHODS: Seventeen infertile women with a self-reported history of exposure to DES in utero, attending the IVF unit at Massachusetts General Hospital (MGH) for assisted reproductive technology (ART), underwent 27 IVF cycles. Analysis of the outcome of IVF including implantation and ongoing pregnancy rates was performed. The data were compared with results from a group of 20 infertile patients with idiopathic infertility undergoing 27 IVF cycles at MGH during the same period. The patients in the two groups were matched for age, basal day 3 levels of follicle stimulating hormone and serum estradiol, and the number and quality of embryos transferred. RESULTS: The response to controlled ovarian hyperstimulation was comparable in the two groups. Significantly lower implantation and ongoing pregnancy rates following IVF and embryo transfer were seen in the utero DES exposed group compared to the control patients. CONCLUSIONS: Infertile patients with a history of in utero exposure to DES exhibit a significantly impaired implantation rate following IVF, and the outcome of ART remains poor. PMID- 9401870 TI - Endogenous LH surge versus hCG as ovulation trigger after low-dose highly purified FSH in IUI: a comparison of 761 cycles. AB - PURPOSE: The results obtained with a protocol consisting of ovarian stimulation with low doses of highly purified FSH (FSH HP), administration of a GnRH analogue to induce an endogenous surge of gonadotropins, and IUI were evaluated. These results were compared with those seen with similar FSH stimulation and hCG administration followed by IUI. METHODS: Three hundred sixty-four patients scheduled for IUI, after inclusion in a total of 345 FSH HP/GnRH-stimulated cycles and 416 FSH HP/HCG-stimulated cycles, were studied. The stimulation protocol consisted of daily subcutaneous injection of 75 IU of FSH HP from day 3 or 5 of the cycle, depending on the duration of the spontaneous cycle. hCG was administered on days 0, +2, and +5 to support the luteal phase. Monitoring was conducted using circulating estradiol levels and vaginal ultrasonography. Administration of two s.c. doses of leuprolide acetate (LA) or 7500 IU of i.m. hCG when at least one 18-mm-diameter follicle was seen and estradiol levels reached 120 pg/ml per follicle with a diameter > or = 16 mm. Intrauterine insemination was with semen capacitated by swim-up, thawed at room temperature if previously frozen. RESULTS: The ovulation rate was 99.28 after hCG and 99.23 with LA. No significant differences were seen between the estradiol and progesterone levels of both groups or in the estradiol/progesterone ratio. The duration of the luteal phase was similar in both groups. Pregnancy rates per cycle were 17.31% (hCG) and 27.25% (LA), respectively (P = 0.0007), and abortion rates 22.22% (hCG) and 24.47% (LA), respectively. No cases of ovarian hyperstimulation were seen. CONCLUSIONS: After FSH HP administration according to a low-dose protocol, the use of LA to trigger a gonadotropin surge as a means of inducing ovulation in FSH stimulated women could be a good alternative to improve the results and prevent ovarian hyperstimulation in IUI cycles. PMID- 9401871 TI - Incomplete androgen and progesterone suppression following midluteal GnRHa prior to controlled ovarian hyperstimulation for IVF-ET. AB - PURPOSE: We aimed to determine if midluteal GnRH agonist (GnRHa) use prior to controlled ovarian hyperstimulation (COH) results in uniform progesterone and androgen suppression and whether elevations of these hormones occurring early in follicular development may adversely effect the outcome of IVF-ET. METHODS: Forty four COH cycles using midluteal GnRHa were evaluated. Serum gonadotropins (LH and FSH) and gonadal steroids (E2, A, P4, and T) were measured after 10 days of GnRHa administration [cycle day 31 (CD 31)] and again on the day of hCG administration, following COH. Cycle outcomes evaluated were the number of oocytes retrieved, morphologic grade, fertilization, implantation, pregnancy, and spontaneous abortion rates. RESULTS: Endogenous serum FSH was uniformly suppressed (6.32 +/- 0.47 IU/L) on CD 31, however, LH was not (23.76 +/- 0.76 IU/L). Five and four tenths percent of cycles demonstrated low-level P4 elevations (> or = 0.9 ng/ml), 24.4% demonstrated serum androstenedione levels > or = 600 pg/ml, and 39% of cycles were characterized by serum T levels > or = 200 pg/ml despite evidence of E2 suppression (< or = 30 pg/ml) and the absence of follicular growth by sonography. LH levels were not predictive of incomplete P4 or androgen suppression. Elevations of either P4, A, or T occurring early in the follicular phase were not found to correlate with an impairment in clinical cycle outcome. CONCLUSIONS: Midluteal GnRHa use prior to COH may result in incomplete suppression of circulating progresterone and androgens. However, these relative elevations, occurring early in the development of the follicular cohort, did not appear to affect IVF cycle outcome adversely. PMID- 9401872 TI - Effects of cryopreserved semen quality and timed intrauterine insemination on pregnancy rate and gender of offspring in a donor insemination program. AB - PURPOSE: Our purpose was to study the relationship among cryopreserved donor semen quality, pregnancy rates, and preconception sex selection after intrauterine insemination. METHODS: We reviewed the records of the 203 women in our donor insemination program from 1987 to 1994 who became pregnant after more than one insemination cycle and had no female-factor infertility. They were categorized according to the number of cycles required for pregnancy. Semen samples from 54 donors were analyzed before freezing and after thawing. Specimens resulting in pregnancy were compared to specimens from the same donor that did not. Semen characteristics were compared to gender of the child. RESULTS: Two hundred fifty two-women became pregnant of the 422 who were enrolled. The pregnancy rate per cycle was 13%. Semen quality was not related to pregnancy outcome or offspring gender. However, more male children (101 vs 83) were born. CONCLUSIONS: Semen characteristics in good-quality cryopreserved donor semen do not affect pregnancy rate or offspring gender. PMID- 9401873 TI - Creatine kinase level and lipid peroxidation rate in human spermatozoa from patients with cancer. AB - PURPOSE: The present study assessed whether the poor semen quality in patients with cancer results from the inhibition of sperm maturation as indicated by creatine kinase or from increased oxidative stress as assessed by lipid peroxidation of the sperm membrane. METHODS: Cryopreserved semen specimens from patients with testicular (n = 10) and nontesticular (n = 12) cancer and normal healthy donors (n = 14) were analyzed for lipid peroxidation and creatine kinase levels. RESULTS: The levels of creatine kinase and malonaldehyde did not differ among testicular or nontesticular patients with cancer or normal healthy donors. CONCLUSIONS: Poor semen quality in testicular and nontesticular patients with cancer is not related to creatine kinase or lipid peroxidation levels; it may be related to other factors. PMID- 9401875 TI - FISH and the paediatrician. AB - Cytogenetics with banding techniques has, since the 1970s, identified patients with chromosome abnormalities and has contributed enormously to the understanding of phenotype-karyotype correlations. However, one chromosome band could contain 20-50 genes. Fluorescence in situ hybridization (FISH) bridges the gap in the area between the resolution obtained by conventional chromosome studies and purely DNA studies. Fluorescence in situ hybridization provides paediatricians with the ability to delve more deeply into the aetiology of congenital abnormalities in children. This annotation aims to clarify the current applications of FISH in paediatric practice. PMID- 9401874 TI - Culture with Matrigel inhibits development of mouse zygotes. AB - PURPOSE: It was reported that Matrigel improved hatching of mouse blastocysts produced in vitro from F1 hybrid-derived zygotes. We investigated whether Matrigel would be similarly beneficial with outbred strain-derived embryos, which exhibit a "two-cell" block similar to the developmental blocks of other species. METHODS: Mouse embryo development was assessed with or without Matrigel in KSOM medium, which supports the development of blocking strain zygotes in vitro, and in human tubal fluid (HTF) medium, which normally does not but which is used for human IVF. RESULTS: Matrigel severely inhibited the development of zygotes to blastocysts in KSOM and did not improve culture in HTF. There was no effect on development from the two-cell stage. We were not able to replicate the previous finding of Matrigel's beneficial effect on hatching of F1-derived zygotes. CONCLUSIONS: Matrigel may be a deleterious addition to embryo culture or coculture systems. PMID- 9401876 TI - Psychosocial impact of chronic illness in children. AB - Chronic illness is common in childhood and is associated with an increased risk of psychological difficulties in the child. Current research is focused on the identification of specific risk and protective factors that may predict psychological and health outcomes. The challenges faced by physicians caring for a child with chronic illness are described and contrasted with the medical role in treating acute illness. The child's adaptation to illness is discussed in a developmental framework and positive and maladaptive family responses are identified. It is suggested that chronic illness and/or its treatment may compromise intellectual development and academic progress. PMID- 9401877 TI - Paediatric HIV update. AB - Less than half of the paediatric HIV infections recorded in Australia have resulted from perinatal transmission, but in recent years this has been the predominant mode of infection. There are 136 infants who are known to have been exposed perinatally to HIV in Australia: 49 of these are infected. Caesarean section is thought now not to reduce the risk of perinatal transmission (PNT); rather, the risk increases with duration of membrane rupture and rises rapidly after 4 h of membrane rupture. However, no data exist to show that interventions to expediate delivery after membrane rupture reduce the risk of PNT. Data such as these suggest that the majority of perinatal infections (probably about 60%) occur close to the time of delivery. While the overall risk of PNT for non-breast fed infants is approximately 20-25%, the risk of infection for the infant is considerably increased when there is evidence of increased maternal viral burden. Advanced maternal disease predicts that if the infant is infected there is more likely to be early progression of HIV than is the case for the less frequently infected infants of mothers who are asymptomatic. Bottle feeding may prevent infection of 10% of children exposed perinatally. Use of zidovudine by the mother in the third trimester and i.v. zidovudine during labour, followed by oral zidovudine for the infant for 6 weeks can reduce the PNT rate by two thirds, to about 8%. Approximately 3% of uninfected infants with perinatal HIV exposure may be found to be transiently virus positive but eventually become antibody negative and thus appear to have eliminated the virus. The risk of Pneumocystis carinii pneumonitis (PCP) cannot be predicted on the basis of CD4 count and it is recommended that all children of infected mothers commence PCP prophylaxis around the age of 6 weeks-2 months and continue that therapy until the age of 12 months or until it becomes clear that the infant is uninfected. The cumulative risk of AIDS increases rapidly during the first year of life to about 20%, then more slowly at a rate of about 2 or 3% a year. The shape of this curve reveals the bimodal progression of HIV disease in children. About 15-20% of children rapidly develop a severe immune deficiency, opportunistic infections and, in most cases, encephalopathy. There is a very high morbidity rate in this group of children, most of whom die before the age of 3 or 4 years. In contrast, 80-85% of children only become immunodeficient after a relatively long period, which is similar to or perhaps even longer than that in adults. Recent studies indicate that zidovudine antiviral monotherapy is no longer appropriate. While no clear alternative to monotherapy has emerged most would, wherever possible, commence antiretroviral therapy with a combination of two or three drugs including zidovudine plus didanosine or lamivudine. If a third drug is used it would probably be a protease inhibitor. PMID- 9401878 TI - Social skills training for primary school children: a 1-year follow-up study. AB - OBJECTIVE: To evaluate the effectiveness of the Rochester Social Problem Solving Program to reduce emotional and behavioural problems amongst primary school children. METHODOLOGY: Children in years 3 and 4 at primary school were assessed prior to receiving the program, immediately after the program and 1 year after the program. At each assessment, the functioning of the children who received the program was compared to the functioning of children enrolled in years 3 and 4 at a comparable school who did not receive the program. RESULTS: The program improved the ability of children to cope with potentially difficult social situations. However, the program did not reduce the prevalence of teacher reported or mother-reported childhood emotional and behavioural problems. CONCLUSIONS: School-based social skills programs may be more effective in reducing childhood emotional and behavioural problems if they include components which focus specifically on childhood behaviour problems as well as components focusing on social skills and peer relationships. PMID- 9401879 TI - Discrimination of attention deficit hyperactivity disorder by the continuous performance test. AB - OBJECTIVE: To investigate the Continuous Performance Test in discriminating a group of 56 attention deficit hyperactivity disorder (ADHD) children from 56 school children individually matched for age, sex and social class. METHODOLOGY: The children all completed the Continuous Performance Task (CPT). The mothers and teachers completed a Conners Parent-Teacher Rating Scale for the clinic children. RESULTS: The ADHD sample was selected so that the average IQ was 99.8 to match the school sample. A non-parametric discriminant function showed that the subtests of the CPT that best discriminated ADHD were age-normalized errors of commission (NCPTC) and age-normalized mean reaction time (NMNRT). CONCLUSION: Optimal use of the CPT for discrimination of ADHD should include age normalization and mean reaction time to targets. Further evoked potential studies may show brief cortical events involved in reaction time over the course of the CPT, and the processes involved in behavioural control. PMID- 9401880 TI - Clinical and endoscopic predictors of histological oesophagitis in infants. AB - OBJECTIVE: To define the earliest age at which histological changes can be used to diagnose oesophagitis and to determine the relationships between clinical, endoscopic and histological features of oesophagitis in infants. METHODOLOGY: The case records and biopsies of 113 infants aged 2-18 months with clinically significant gastro-oesophageal reflux (GOR), undergoing oesophagoscopy between 1978 and 1994 were retrospectively reviewed. The biopsies were independently evaluated and graded by two pathologists. RESULTS: Forty-five cases (40%) had histological oesophagitis but only 16 (14%) had abnormal endoscopic findings (excluding erythema). Endoscopy was found to be highly specific (93%) for histological oesophagitis but lacked sensitivity (25%). Irritability was inversely related to the presence of endoscopic abnormalities, and there was poor correlation between symptoms and histological changes with only haematemesis showing a statistically significant association with histological abnormalities (P = 0.033). Intraepithelial lymphocytes were the earliest of the histological features noted and were present before 4 months of age. The numbers of intraepithelial eosinophils and lymphocytes and the presence of papillary elongation all increased with age. CONCLUSIONS: The presence of oesophagitis is difficult to predict on the basis of symptoms. The presence of intraepithelial lymphocytes is the earliest histological change to be seen in infants with GOR, and can develop before 4 months of age. Oesophagoscopy without biopsy is unreliable in the diagnosis of oesophagitis in infants. PMID- 9401881 TI - Improvement in outcome of children with Wilms' tumour in South Australia over the last 30 years. AB - OBJECTIVE: To evaluate the outcome of children with Wilms' tumour over the last 30 years in South Australia. To compare the outcome of children treated before and after 1982, when standard treatment protocols were introduced. METHODOLOGY: Management approaches, survival rates and side-effects of treatment were identified from case notes. Pathology slides were reviewed to ensure all children were correctly diagnosed with Wilms' tumour. RESULTS: Children treated for Wilms' tumour prior to 1982 had an overall survival rate of 54%. Since 1982 there has been a significant improvement in outcome and the current survival rate is now 85%. Children treated since 1982 have also experienced fewer treatment related side-effects than children treated prior to 1982. CONCLUSIONS: There has been substantial improvement in survival from childhood Wilms' tumour over the past 30 years in South Australia. This is likely to be due to a combination of factors including standardization of treatment, tailoring of treatment to stage and histology, improved perioperative care, enhanced radiological techniques and higher levels of collaboration between relevant specialists. PMID- 9401882 TI - Control of temperature during newborn transport: an old problem with new difficulties. AB - OBJECTIVE: To explore any changes in temperature control during neonatal emergency inter-hospital transport between 1977 and 1996. METHODS: Records were reviewed of all infants undergoing emergency transfer by the statewide Victorian Newborn Emergency Transport Service (NETS). Per axillary temperatures were recorded prospectively on arrival of transport team and at conclusion of transfer for all infants. RESULTS: The rate of hypothermia (< 36.0 degrees C) when NETS reached the infant has decreased overall (22% in 1977-79 to 7% in 1995-96) and for all weight groups; although in 1995-96 hypothermia was present in 36% of infants less than 1000 g when NETS arrived. The rate of hypothermia (< 36.0 degrees C) at the end of the transfer has remained at 3% overall for many years. The rate of hyperthermia at both times has increased significantly overall (12% in 1977-79 to 24% in 1995-96 on NETS arrival, 4%-19%, respectively at end of transfer) and for all weight groups except infants less than 1000 g. The range of abnormal temperatures has not substantially changed over time. CONCLUSION: There has been significant improvement in avoidance of hypothermia and cold stress amongst infants requiring emergency neonatal transport from 1977 to 1996. However, in order to improve the number of infants transferred who achieve a temperature in the normal range the need to avoid hyperthermia is highlighted. Infants who require incubator care for optimal medical management require continual monitoring of temperature and review of environmental conditions to optimise the conditions both prior to and during transport. PMID- 9401883 TI - Chronic oxygen dependency in infants born at 24-32 weeks' gestation: the role of antenatal and neonatal factors. AB - OBJECTIVES: To study the incidence of chronic oxygen dependency (COD) among ventilated survivors born at 24-32 weeks gestation from 1986 to 1994 and to identify antenatal and neonatal factors that may have changed with time; and to identify antenatal and neonatal factors that could contribute to the development of COD in infants born at 24-32 weeks gestation using a case control model. METHODOLOGY: Infants born at 24-32 weeks gestation in one tertiary referral centre between 1986 and 1994 and admitted to the neonatal intensive care unit for respiratory support were studied. Data accumulated prospectively since 1986 in survivors of ventilation were analyzed to identify antenatal and neonatal factors that could have changed with time. The cohort of infants who developed COD were matched for gestation and time of birth with a control group of infants who did not have COD. Significant factors that could have contributed to the development of COD were identified using forward logistic regression analysis. RESULTS: The number of mothers admitted for threatened premature labour (TPL), and pregnancy induced hypertension decreased with time while the use of antenatal steroids and maternal antibiotics increased. More infants were delivered by Caesarean section during the later years. There was an increase in neonatal septicaemia with time while there were decreases in hyaline membrane disease, pneumothorax, pulmonary interstitial emphysema, use of high peak inspiratory pressure (PIP) and high inspired oxygen. The incidence of COD decreased. The case controlled study revealed a significant positive association between COD and male gender, birthweight less than the 10th percentile for gestation, PIP over 30 cm H2O, septicaemia and significant patent ductus arteriosus (PDA) requiring indomethacin. There was a negative association with TPL. CONCLUSIONS: Further decrease in COD can be achieved only if septicaemia, PDA and the use of high PIP can be avoided. The most effective way of reducing the incidence of COD is by reducing the incidence of prematurity. PMID- 9401884 TI - The seasonal distribution of infant deaths by age: a comparison of sudden infant death syndrome and other causes of death. AB - OBJECTIVE: To examine the possibility that among deaths in infancy the increase in the winter/summer ratio with increasing age is not peculiar to sudden infant death syndrome (SIDS). METHODOLOGY: Details of the winter (December February)/summer (June-August) ratio among deaths in neonates (< 28 days) and post neonates dying in the United States of America between 1979 and 1990 were abstracted from published statistics. The primary causes of death were classified according to the ninth Revision of the International Classification of Diseases. RESULTS: For every non-traumatic cause of death including SIDS, the winter/summer ratio was higher among postneonates than neonates. This was not seen for deaths due to trauma. Cases of SIDS and deaths due to infection had the highest ratios in both age categories. Causes of death occurring predominantly in the neonatal period (e.g. anencephaly) had the lowest overall ratios. CONCLUSIONS: Neither the greater number of SIDS cases in the winter, nor the increasing winter/summer ratio with increasing age is unique to SIDS. PMID- 9401885 TI - Primary course immunogenicity and reactogenicity of a new diphtheria-tetanus whole cell pertussis vaccine (DTPw). AB - OBJECTIVE: To establish safety, immunogenicity, and batch stability of a reformulated whole cell pertussis based diphtheriatetanus-whole cell pertussis (DTP) vaccine (nDTPw) compared to the currently marketed Australian DTPw vaccine (Triple Antigen) in a three dose 2, 4 and 6 month primary immunization course. Reformulation was necessary to make the DTPw vaccine suitable for combination with hepatitis B and Haemophilus influenzae b vaccines. METHODS: Double blind randomized controlled trial in suburban Melbourne in 812 healthy infants recruited through maternal and child health centres, of whom 208 received Triple Antigen and 604 received nDTPw. RESULTS: Results for both reactogenicity and immunogenicity were similar and were not significantly different for the three batches of nDTPw. No new, serious, or unexpected adverse effect was recorded. Nearly twice as many nDTPw infants experienced no general reaction to the third dose (18%) compared to Triple Antigen (11%, P = 0.06). An elevated temperature (> or = 38 degrees C axillary) occurred in about three out of 10 babies overall, with rates being slightly higher for both vaccines after the second vaccination. Local reaction rates were significantly less common for nDTPw on days 2 and 3 following each of the three vaccinations. After dose three, 30% of nDTPw subjects experienced no local reaction compared to 20% of Triple Antigen subjects (P = 0.015, 95% Cl on difference 2%, 19%). Swelling after doses one and three occurred in 30% and 24% of Triple Antigen subjects, compared to 23% (P = 0.07, 95% Cl diff 0%, 13%) and 14% (P = 0.017, 95% Cl diff 2%, 17%) of nDTPw subjects. Tenderness after doses two and three occurred in 80%, and 78% of Triple Antigen subjects, compared to 71% (P = 0.04, 95% Cl diff 1%, 17%) and 68% (P = 0.025, 95% Cl diff 2%, 19%) of nDTPw subjects. There was a significantly higher post immunization diphtheria antitoxin GMT (2.73 IU/mL) for Triple Antigen compared to nDTPw (1.89 IU/mL; P = 0.02), although no subject in either vaccine group had a tetanus or diphtheria antibody titre less than six times the protective level of 0.01 IU/mL following immunization. The nDTPw post immunization GMTs were significantly higher for Agg2, Fha, and pertactin compared to Triple Antigen geometric mean titres (GMTs). CONCLUSION: nDTPw is a safe and immunogenic vaccine when compared to Triple Antigen. The reformulated vaccine is an acceptable replacement for the currently marketed formulation, and for evaluation as a component of future combination vaccines. PMID- 9401886 TI - Incidence of apnoea and bradycardia in preterm infants following DTPw and Hib immunization: a prospective study. AB - OBJECTIVE: To evaluate the incidence and severity of apnoea and bradycardia in hospitalized preterm infants following immunization at 2 months of age, and identify risk factors. METHODOLOGY: A prospective study of 98 preterm infants, of gestational age 24-31 weeks, immunized at approximately 2 months post natal age with diphtheria-tetanus-whole cell pertussis vaccine (DTPw) in the neonatal intensive care unit (NICU) at King George V Hospital Sydney. Half the infants also received Haemophilus influenzae type b conjugate vaccine (Hib) simultaneously. All infants were monitored for apnoea and bradycardia in the 24 h periods pre- and post immunization. RESULTS: Only one infant had apnoea and/or bradycardia pre-immunization compared with 17 post immunization. For 12 infants these events were brief, self-limiting and not associated with desaturations (oxygen saturation < 90%). However, for five infants (30%) these events were associated with oxygen desaturation and two of these infants required supplemental oxygen. The group that had apnoea and/or bradycardia and the group that did not were not significantly different in terms of gestational age, birth weight and other variables. Infants who received Hib together with DTPw were less likely to have apnoea and/or bradycardia than those given DTPw alone. CONCLUSION: When considering immunization for preterm infants, the benefits of early immunization must be balanced against the risk of apnoea and bradycardia. We recommend that the cardio-respiratory function of hospitalized infants born at less than 31 weeks gestation be monitored for 48 h post immunization. PMID- 9401887 TI - Young Malaysian children with lower respiratory tract infections show low incidence of chlamydial infection. AB - OBJECTIVE: The incidence of Chlamydia pneumoniae and Chlamydia trachomatis infection was studied among infants and young children admitted to hospital for the management of lower respiratory tract infections, over a 12 month period. METHODOLOGY: Respiratory secretions were examined for chlamydiae by cell culture, enzyme-linked immunosorbent assay and polymerase chain reaction-enzyme immunoassay. Sera were tested by micro-immunofluorescence for chlamydial IgG, IgM and IgA. Other bacterial and viral pathogens were also looked for by standard cultural and serological methods. RESULTS: Of 87 patients aged 2 months-3 years, an aetiologic diagnosis was made in 41 (47.1%). C. pneumoniae and C. trachomatis were each detected in 1 (1.2%) of the patients. Among common bacterial pathogens, Haemophilus influenzae (13.8%) and Streptococcus pneumoniae (8.1%) were the most frequently identified. Respiratory viruses and elevated Mycoplasma pneumoniae antibodies were found in 10.3% and 9.1% of patients, respectively. CONCLUSION: Chlamydiae are infrequent causes of community-acquired acute lower respiratory tract infections in infants and very young children in Malaysia. PMID- 9401888 TI - The health of a group of young Australians in a New South Wales juvenile justice detention centre: a pilot study. AB - OBJECTIVE: To consider the health profile of a sample of young, largely male Australians as assessed on their admission to a New South Wales Juvenile Justice Detention Centre. METHOD: A retrospective analysis of primary care nurse health records for 100 sequential admissions. RESULTS: Of the 97 males and three females (mean age = 15.9 years), 30 were Aboriginal and 39 did not live with either parent at the time of admission. Respiratory illness, such as bronchitis and asthma were common. These diagnoses were overshadowed by histories of significant physical injury. The sample was at high risk of sexually transmitted disease. Forty-six per cent had prior contact with a mental health professional, 26% reported they had thought of suicide and 9% reported having attempted suicide. There was a high prevalence of substance abuse. CONCLUSION: The health of these young Australians is at risk from every perspective. Improving the quality of their health assessments is an important issue for the clinicians who attend them as individuals and for policy makers who aim to reduce the considerable social and economic cost of juvenile crime. The discussion of these results from one centre has revealed opportunities to make such improvements. There is a need for a gathering of expertise to address the issue, preferably on a national basis. PMID- 9401889 TI - Acne in Victorian adolescents: associations with age, gender, puberty and psychiatric symptoms. AB - OBJECTIVES: This study aimed to examine the associations between the frequency and severity of self-reported acne and age, gender, puberty and psychiatric symptoms in Victorian adolescents. METHODOLOGY: A sample of secondary schoolchildren in Victoria, Australia were surveyed using a computerized questionnaire. Developmental and psycho-social factors associated with acne were recorded and analysed using logistic regression. RESULTS: The Victorian Adolescent Health Survey (1992) recorded the frequency and severity of self reported acne in 2491 students. Frequency of acne increased with age and pubertal development. For females commencement of menstruation was associated with increased frequency of acne. Asian born male students were less likely to report acne than Australian born males. Acne severity was coded into mild (students reporting acne sometimes on back or face) and moderate (students reporting acne often on face or back). Students reporting moderate acne were more likely to report a high level of psychiatric symptoms and were in the later stages of puberty. CONCLUSIONS: This study confirms an association between the frequency and severity of self-reported acne and stage of pubertal development. It showed also that students reporting moderate acne were more likely to report psychiatric symptoms of depression and anxiety. PMID- 9401890 TI - Trends in the health burden due to urinary tract infection in children in Australia. AB - OBJECTIVE: To estimate the health burden of urinary tract infection in children less than 15 years of age in Australia and to ascertain whether any significant change has occurred during the past decade. METHODOLOGY: The number of children less than 15 years of age who were admitted in New South Wales for urinary tract infection between 1981 and 1994 was ascertained from the Department of Health, and age and sex specific incidence rates were calculated using Australian Bureau of Statistics population data. Costs for inpatient care were calculated using the cost weights from Australia National Disease Related Groups Version 3 for urinary tract infection (DRG 577). The frequency of the four most commonly requested renal tract imaging procedures in children following urinary tract infection and which qualified for Medicare reimbursement were obtained from the Health Insurance Commission for 1984-1994: micturating cystourethrography, intravenous urography, renal ultrasonography, and nuclear medicine renal studies. RESULTS: There were 1203 children who were admitted with urinary tract infection in New South Wales in 1994, at an estimated cost of $A1.6 million. Since 1981, the age standardized annual incidence of urinary tract infection requiring hospitalization has increased from 0.5 to 0.9 per 1000 children, largely because of an increase in the number of young children admitted (from 0.6 to 2.0 per 1000 children less than 5 years of age). In 1994, 46,230 non-inpatient renal imaging procedures were undertaken in children under 15 years of age at a cost of $A5.3 million. CONCLUSIONS: Urinary tract infection is an important and increasing health problem for Australian children, particularly for preschool children. Whether this represents a true increase in the incidence of urinary tract infection or improved diagnosis and more intensive management is not possible to establish with this study design. Prospective population based studies are required to assess more completely the frequency with which urinary tract infection occurs in children and any changes that may be occurring. PMID- 9401891 TI - Limitations of school entry immunization certificates. AB - OBJECTIVE: To compare the immunization certificate submission rate for kindergarten school children on the Central Coast of New South Wales for 1994 and 1995; to contact individual parents of children with incomplete or missing immunization certificates by issuing a personalised immunization reminder letter, and measuring their response. METHODOLOGY: Seventy Central Coast schools were surveyed. Immunization documentation for 4177 kindergarten enrolements in 1995 were assessed. Children with incomplete or unknown immunization status were identified. Immunization reminder letters were sent by the schools to parents of the children that were identified. The school recorded any additional immunization certificates submitted. RESULTS: In 1995, 79% of kindergarten children submitted immunization certificates, unchanged from the previous year. Immunization reminder letters were sent to 856 families of children with no certificate and 215 families of children with an incomplete certificate. A further 263 immunization certificates were submitted, representing a 25% response rate. This intervention increased the percentage of kindergarten children with a certificate from 79% to 86%. CONCLUSIONS: Education of school staff during the 1994 immunization survey did not change the overall immunization certificate submission rate. Specifically targeting parents through schools can be an effective way of increasing immunization certificate submission. PMID- 9401892 TI - Vascular air embolism: a rare complication of nasal CPAP. AB - A preterm infant developed bilateral tension pneumothoraces and extensive vascular air embolism 6 h after being commenced on nasal continuous positive airway pressure (CPAP). Neonatal clinicians should be aware that catastrophic vascular air embolism could occur in infants receiving nasal CPAP, a modality of respiratory support conventionally considered non-invasive and 'safe'. PMID- 9401893 TI - Epidural haematoma and stroller-associated injury. AB - We present a severe case of head injury in an infant associated with stroller use and review similar reports from the literature. A case report and a literature search of the Medline database from 1966 to 6/1996 using the terms 'perambulator', 'parm', 'stroller', or 'baby carriage'. Reports in English describing injuries associated with their use are reviewed. We report a case of epidural haematoma in a 10 months old girl who sustained the injury after falling from a stroller. Safety harnesses were not worn during the incident. There was no skull fracture. Complete recovery followed surgical evacuation of the blood clot. Five reports describing stroller-related injuries were found in the English literature. Most injuries were mild. Three cases of death were reported of which two were classified as child abuse. The prevalence of unintentional stroller associated injury is not clear. Mild injury, mostly to the head region, is probably common. Life-threatening injuries are rare but these are potentially preventable if strollers are properly designed and safety recommendations are followed. PMID- 9401894 TI - Haemoptysis in healthy children due to unsuspected foreign body. AB - Haemoptysis in otherwise healthy children is an uncommon event. Two cases of massive haemoptysis, subsequently requiring lobectomy, are discussed. In each case, foreign vegetable matter was identified despite previously normal bronchoscopy and minimal changes on chest radiograph. PMID- 9401895 TI - A case of kernicterus in New Zealand: a predictable tragedy? AB - A case of neonatal kernicterus due to glucose-6-phosphate dehydrogenase deficiency (G6PD) is described. Diagnosis was delayed as the primary healthcare attendant had no knowledge of this condition and its potential to cause rapidly escalating levels of bilirubin and as she was reassured by the lack of signs of systemic illness or anaemia. The baby has been left deaf, blind, intellectually handicapped, epileptic and paralysed due to athetoid cerebral palsy. The re organization of perinatal care in New Zealand, which has led to neonates sometimes being managed solely by primary healthcare attendants with minimal training in paediatrics may have increased the risk of a late diagnosis of potentially devastating diseases such as this. PMID- 9401896 TI - Yellow nail syndrome in infancy. AB - This report describes an infant with clinical features consistent with the yellow nail syndrome (YNS), a rare autosomal dominant disorder. He presented at birth with congenital lymphoedema and was referred at 6 months of age for investigation of recurrent cough and wheeze. He had clinical and radiological evidence of bilateral pleural effusions and a pericardial effusion. Following a lung biopsy and pericardial window these were shown to be manifestations of his lymphatic abnormality. He also had persisting middle ear effusions causing conductive deafness requiring hearing aids and secondary immunodeficiency requiring regular immunoglobulin infusions. PMID- 9401897 TI - Prevalence of obesity and hypercholesterolaemia in children: a cohort study. PMID- 9401898 TI - Efficacy of ipratropium bromide by metered dose aerosol and aerochamber in acute paediatric bronchiolitis. PMID- 9401899 TI - Increase in asthma severity in the west. PMID- 9401900 TI - Chronic illness in adolescents: transfer or transition to adult services? PMID- 9401901 TI - Insulin-dependent diabetes mellitus: a constellation of autoimmune diseases. PMID- 9401902 TI - Regional variability in the epidemiology of childhood diabetes in Italy. AB - The incidence rates of IDDM in Italy show remarkable variability. Sardinia, a region with the second highest incidence rate in the world, co-exists with other regions with lower rates. We review and compare epidemiologic data on the incidence of childhood-onset IDDM in Italy. papers published from 1980 to 1996 reporting incidence data in Italian areas were found by search of Medline and non indexed Italian journals. The incidence data found cover only 57% of the Italian population. The analysis of our results shows how difficult it is to make a careful study of epidemiology of IDDM in Italy. The RIDI (the Registry for Insulin-dependent Diabetes mellitus in Italy) project started in 1996 according to international guidelines. The aims is to coordinate local IDDM registries, to promote the start of new registries in uncovered areas, and to standardize registration and data collection. PMID- 9401903 TI - Thyroid nodules in childhood and adolescence--thirty years of experience. AB - In the thirty year period between 1966 and 1996, fifty-two patients underwent surgery for thyroid nodules at the Royal Children's Hospital, Melbourne. We aimed to review their presentation, investigation, histology, treatment and to follow up those who had malignant neoplasms. Forty-one of the fifty-two patients presented with a single thyroid nodule. Investigations performed included thyroid function tests (N = 32), thyroid autoantibodies (N = 21), an ultrasound of the thyroid (N = 26) and 99mTechnetium scanning (N = 32). Thirty-five of the neoplasms were benign, the follicular adenoma (N = 16) being the most common. Seventeen patients had malignant neoplasms, seven of whom had papillary and seven of whom had follicular carcinoma. Three patients had medullary carcinoma of the thyroid. Nine of the seventeen patients with thyroid malignancy received post operative 131I treatment. At the time of this review, all patients were living. PMID- 9401904 TI - Growth pattern and age at menarche of obese girls in a transitional society. AB - Childhood obesity is an increasing problem in a transitional society such as Thailand. To study physical growth and puberty in obese children, a cross sectional survey of growth and age at menarche was carried out in schoolgirls aged between 8 and 16 years old. The 3,120 girls were divided into two groups based on weight-for-height criteria. Girls with weight-for-height between 80 and 120% were classified as normal stature (2,625; 84.1%) and those more than 120% were obese (495; 15.9%). Using probit analysis, age at menarche in obese girls was 0.9 year earlier than normal stature girls (11.5 years vs 12.4 years). At age 12, obese girls were reaching menarche 2.8 times more when compared with the normal stature girls. In terms of growth pattern, obese girls were taller and grew faster during the prepubertal period, and then reached their final height earlier than the normal stature girls (13 years vs 15 years). The final height in obese girls was significantly shorter (153.0 cm and 155.0 cm, p = 0.01). We conclude that: 1) obese girls grow faster, have earlier menarche and then stop growing earlier, and 2) obese girls tend to be shorter as adults, compared with normal stature girls. PMID- 9401905 TI - The GH-releasing effect of Hexarelin, a synthetic hexapeptide, in newborns is lower than in young adults. AB - The aim of the present study was to verify the GH-releasing effect of Hexarelin, a synthetic hexapeptide, in newborns who are known to have GH hypersecretion likely due to hyperactivity of GHRH-secreting neurons while somatostatinergic activity seems not fully operative. We studied in 6 newborns (NB, 2.5 +/- 2.1 days), 12 prepubertal children (PC, 9.8 +/- 0.45 yr) and 12 young adults (YA, 28.2 +/- 0.2 yr) the GH response to Hexarelin (HEX, 2 micrograms/kg i.v.) compared to that observed after GHRH (1 microgram/kg i.v.) in 6 NB (4.2 +/- 0.4 days), 12 PC (9.9 +/- 0.6 yr) and 12 YA (31.0 +/- 1.3 yr). GH levels were assayed basally and 30 and 60 min after drug administration. In NB, mean (+/- SEM) basal GH levels were higher while IGF-I levels were lower than those recorded in PC and YA (GH: 34.8 +/- 1.9 vs 2.8 +/- 0.4 vs 1.4 +/- 0.4 micrograms/l, p < 0.0006; IGF I: 36.3 +/- 1.9 vs 152.0 +/- 11.5 vs 175.8 +/- 15.3 micrograms/l, p < 0.0007); in the last two groups GH and IGF-I levels were similar. The mean delta GH peak after HEX in NB (32.8 +/- 4.7 micrograms/l) was similar to that in PC (34.6 +/- 4.3 micrograms/l) and lower (p < 0.01) than that in YA (56.2 +/- 7.4 micrograms/l). Delta GH peak after GHRH in NB (60.1 +/- 1.5) was higher than those in PC and YA (20.8 +/- 4.8 and 22.8 +/- 3.4 micrograms/l) (p < 0.005 and < 0.002, respectively). In NB, the GH response to HEX was lower (p < 0.005) than to GHRH while in PC and YA the somatotrope response to HEX was higher (p < 0.03 and 0.0004, respectively) than to GHRH. These data demonstrate that the GH-releasing effect of Hexarelin undergoes age-dependent variation being lower in newborns than in young adults, opposite to that observed after GHRH administration. The evidence that Hexarelin releases less GH than GHRH in newborns but not in prepubertal children and in young adults makes unlikely the hypothesis that the GH-releasing effect of this hexapeptide is mediated via endogenous GHRH release. PMID- 9401906 TI - Long-acting gonadotropin-releasing hormone agonists in the differential diagnosis of male precocious puberty. AB - Male sexual precocity is defined as the development of secondary sexual characteristics before 9 years of age. It can be classified as gonadotropin dependent precocious puberty (GnDP) or gonadotropin-independent precocious puberty (GnIP) and sometimes the differential diagnosis between these entities is difficult. To determine whether long-acting GnRH agonists (GnRH-a) are effective in differential diagnosis of male precocious puberty, we measured gonadotropins and testosterone levels 30 days after a single administration of depot GnRH-a (triptorelin, gosereline or leuprolide) in 10 boys with sexual precocity of different etiologies. Testosterone levels 30 days after depot GnRH-a were in the prepubertal range in patients with GnDP but not in GnIP. We conclude that measurement of testosterone levels 30 days after long-acting GnRH-a is effective in the differential diagnosis of male sexual precocity. PMID- 9401907 TI - Serum lipids during parenteral nutrition with a 10% lipid emulsion with reduced phopholipid emulsifier content in premature infants. AB - Fourteen premature infants (range 26 + 0 to 32 + 3), all but two appropriate for gestational age with a mean body weight of 1196 g (range 860 to 2770 g) received a 10% lipid emulsion. This lipid emulsion contained half of the formerly used phospholipid emulsifier concentration reducing the phospholipid/triglyceride ratio to the ratio used for the 20% lipid emulsion (0.06 instead of 0.12). Lipid emulsion was given over a 10 day period commencing at the third day of life with 0.5 g/kg/24 h which was increased daily up to a dose of 2.0-2.5 g/kg/24 h which was reached in all patients at the seventh day of the observation period. During this time mean serum concentrations of cholesterol increased non-significantly from 76.1 mg/dl (SD 33.7) before lipid emulsion to 86.1 mg/dl (SD 36.4) on day seven of the observation period. 13 of the 14 patients (97%) showed no pathological increase of their serum triglyceride concentration during lipid infusion. Mean serum triglyceride concentration increased from 65.3 mg/dl (SD 32.0 mg/dl) before the start of lipid emulsion to 102.6 mg/dl (SD 76.5) on day four (p < 0.05) but with no further significant increase. Lipid emulsions with 10% triglyceride but lower phospholipid content are tolerated without pathological increase in triglyceride or cholesterol serum concentration in the vast majority of premature newborns. PMID- 9401908 TI - Helicobacter pylori infection with parietal cell antibodies in children and adolescents with insulin dependent diabetes mellitus. AB - One hundred and seventy-seven patients with insulin dependent diabetes mellitus (IDDM) diagnosed at the pediatric age were investigated for the presence of gastric parietal cell autoantibodies (PCA). The objective was to evaluate the prevalence of PCA seropositivity and to know whether Helicobacter pylori could be a reason for a higher presence of PCA in IDDM children and adolescents. Twelve of 177 patients (6.77%; confidence interval: 3.1-10.3) had detectable PCA. Gastric pathology was studied in eight of these patients and in seven patients without PCA. Diagnosis of H. pylori infection was made on antral biopsies. None of the patients had an atrophic gastritis. Six of the eight patients with PCA had gastric mucosa colonization by H. pylori and/or chronic gastritis. According to these results, we suggest that H. pylori can be the cause of the presence of PCA positive results in diabetic children and adolescents, and diabetic patients with detectable PCA should be screened for H. pylori. PMID- 9401909 TI - Residential treatment for youngsters with difficult-to-manage insulin dependent diabetes mellitus. AB - A descriptive outcome study of 52 children with insulin-dependent diabetes mellitus (IDDM) admitted to a residential treatment program over 6 years. Criteria for admission included repeated hospitalizations for diabetes-related problems, excessive school absences, and/or familial disruption. Residential treatment included individual, group, and family psychotherapy, diabetes education, and close medical supervision. The design included collection of data before, during and after treatment. Children admitted to the residential unit were compared to a population of pediatric IDDM outpatients from the same geographical area. Treatment was associated with a reduction in diabetes-related hospitalizations, improved school attendance, decreased glycosylated hemoglobin levels, weight gain, individualized insulin changes, improved knowledge about diabetes, and a normalization of attitudes towards this disease. PMID- 9401910 TI - Psychological indications for treatment of tall stature in adolescent girls. PMID- 9401911 TI - Hyperthyroidism in a girl with Down's syndrome. AB - We report an adolescent girl with Down's syndrome, who presented with hyperthyroidism. Autoimmune thyroid disorders can occur in children with Down's syndrome, hypothyroidism developing more frequently. Hyperthyroidism can also be associated with Down's syndrome, and should not be missed. PMID- 9401912 TI - Successful therapy with 3,5,3'-triiodothyroacetic acid (TRIAC) in pituitary resistance to thyroid hormone. PMID- 9401913 TI - Central precocious puberty and chronic renal failure: a reversible condition post renal transplantation. AB - A 3 year-old boy with chronic renal failure associated with prune belly syndrome who developed central precocious puberty is described. He had been maintained on cyclic peritoneal dialysis from age 13 months with creatinine levels of 400-600 mumol/l. Increased linear growth rate probably began at 18 months, and by 38 months of age he had testicular enlargement and pubic hair consistent with Tanner stage 2. Elevated levels of serum testosterone (3.6 nmol/l; normal < 0.7 nmol/l) and luteinizing hormone (LH) (2.8 IU/l; normal < 1.0 IU/l) were demonstrated with a pubertal response to luteinizing hormone-releasing hormone (LHRH) stimulation (peak LH 43.5 IU/l). Other endocrine tests demonstrated hyperprolactinemia (170 micrograms/l; normal 3.4-22 micrograms/l), but normal pituitary-thyroid and pituitary-adrenal functions and normal cranial MR imaging. Despite LHRH-agonist therapy with leuprolide over the next 8 months, he showed an incomplete response with only partial inhibition of basal LH and testosterone levels, and continued significant increments in height standard deviation scores (Ht-SDS) and bone age estimates. However, the sexual precocity appeared fully reversible following a successful living-related renal transplant at age 50 months. Despite discontinuation of leuprolide treatment post-operatively, there was a full reversal of his serum LH and testosterone to a prepubertal profile as well as normalization of the serum prolactin levels. Whereas most boys with chronic renal failure show delayed pubertal development and suppressed linear growth, our patient presents a unique phenomenon of reversible central precocious puberty. The effects of leuprolide therapy in the presence of a uremic milieu and the outcome of successful renal transplantation on sexual precocity are described. PMID- 9401914 TI - Adipsic hypernatremia syndrome in infancy. AB - We report on an infant with chronic hypernatremia due to a congenital defect in osmo-regulation of thirst and the secretion of arginine vasopressin (AVP). A 12 month-old female infant who presented with irritability and signs of dehydration was found to have hypertonic dehydration; plasma osmolality was 430 mOsm/kg B.W. Despite rehydration she remained hypernatremic (serum Na+ 152-158 mEq/l). Lack of signs of thirst led us to the diagnosis of chronic hypernatremia due to adipsia. Laboratory investigation showed: 1. plasma AVP levels were low for plasma osmolality; 2. urine osmolality was normal for plasma AVP, and 3, there was a significant correlation of plasma to urine osmolality (r = 0.72, p < 0.02); however, the slope was markedly reduced indicating partial destruction of the AVP osmoreceptors. PMID- 9401915 TI - Position one analogs of the Saccharomyces cerevisiae tridecapeptide pheromone. AB - Analogs of the Saccharomyces cerevisiae alpha-mating factor [WHWLQLKPGQPMY], in which a variety of residues replaced Trp1 were synthesized and assayed for biological activity and receptor affinity. Analogs containing Gly or Leu or many different aromatic residues in position 1 of the peptide exhibited bioactivity in a growth arrest assay slightly greater than, or equal to, that of the parent pheromone, whereas the Glu1 and Lys1 analogs exhibited significantly lower bioactivity. Analogs with an aromatic replacement at position 1 had 3- to 6-fold lower receptor affinity than the parent peptide, whereas analogs with a hydrophilic residue at the N-terminus exhibited large reductions in receptor affinity with the peptide with Glu in position 1 showing a 120-fold reduction. N alpha-Acetylation had little effect on bioactivity but lowered receptor affinity by 20- to 40-fold. Amidation of the carboxyl terminus resulted in a 10-fold decrease in activity and a 160-fold decrease in receptor affinity. These results indicate that the alpha-factor receptor has a large hydrophobic binding pocket, possibly containing a negatively charged side-chain, which interacts with the N terminus of alpha-factor. The lack of correlation between activity and binding and several analogs suggests that small residues near the N-terminus of alpha factor may be very efficient in triggering isomerization of the receptor to its activated state in the first step of the signal transduction pathway. PMID- 9401916 TI - New mild acid-labile protecting groups for the guanidino function of N alpha fluorenylmethoxycarbonyl-L-arginine in solid-phase peptide synthesis: 10,11 dihydro-5H-dibenzo[a,d]cyclohepten-5-yl, 2-methoxy-10,11-dihydoro-5H dibenzo[a,d]cyclohepten-5-yl and 5H-dibenzo[a,d]cyclohepten-5-yl groups. AB - 10,11-Dihydro-5H-dibenzo[a,d]cyclohepten-5-yl [5-dibenzosuberyl] and 5H dibenzo[a,d]-cyclohepten-5-yl [5-dibenzosuberenyl] groups have been found to be useful protecting groups for the guanidino function of arginine in solid-phase peptide synthesis on Fmoc chemistry. The arginine derivatives (4a,b,c) derivatized with these groups were easily deprotected with mild acid (less than 30 min with 25% trifluoroacetic acid). Tryptophan-containing peptide sequences, two hexapeptides (6) and (8), were synthesized in good yield by mild acid treatment (50% trifluoroacetic acid in 1 h) of the peptide resins (5a,c-f and 7a,c,d) assembled via 4a,b,c using benzotriazol-1-yl-oxy-tris-(pyrrolidino) phosphonium hexafluorophosphate-1-hydroxybenzotriazole mediated coupling. PMID- 9401917 TI - Synthesis and biological activities of head-to-tail cyclic bradykinin analogues of varying ring size. AB - Syntheses of cyclic kinin analogues with different backbone atom numbers are described. Cyclization, by either the O-benzotriazolyl-N,N,N',N' tetramethyluronium tetrafluoroborate/1-hydroxybenzotriazole/diisopropylethyl amine (TBTU-HOBt-DIPEA) or the diphenylphosphoryl azide (DPPA) procedure of linear peptides prepared by the solid-phase method based on the g-fluorenyl methyloxycarbonyl chemistry, was used for preparing cyclo-Gly-Ile-Ile-Gly bradykinin, cyclo-Lys-kallidin (cyclo-Lys-Lys-bradykinin) and cyclo-des Arg bradykinin. Peptides were characterized by amino acid analysis, optical rotation, analytical high-performance liquid chromatography and matrix-assisted laser desorption ionization-time flight mass spectrometry. Pharmacological experiments showed that cyclo-Gly-Ile-Ile-Gly-bradykinin (39 backbone atoms) and cyclo-Lys bradykinin (30 backbone atoms) are about equipotent, when tested on the relaxation of the isolated rat duodenum preparation. The potency of cyclo-des Arg bradykinin is at least three orders of magnitude lower. The potency of cyclo-Lys Lys-bradykinin (33 backbone atoms) is one tenth the activity of bradykinin but about 10 times higher than the potency of the above-mentioned cyclokinins and makes the latter analogue the most potent end-to-end cyclic analogue known currently. The present results, in agreement with data from earlier reports, seem to indicate that the enhancement of the number of backbone atoms in the cyclic kinins first increases and subsequently decreases the potency, whereas a reduction in the atom number from 27 to 24 causes a dramatic decrease in potency. PMID- 9401918 TI - Conformational investigation of alpha,beta-dehydropeptides. VIII. N-acetyl alpha,beta-dehydroamino acid N'-methylamides: conformation and electron density perturbation from infrared and theoretical studies. AB - The Fourier transform infrared spectra are analyzed in the regions of Vs(N-H), amide I, amide II and Vs(C alpha = C beta) bands for a series of Ac-delta Xaa NHMe, where delta Xaa = delta Ala, (Z)-delta Abu, (Z)-delta Leu, (Z)-delta Phe and delta Val, to determine the predominant solution conformation of these alpha,beta-dehydropeptide-related molecules and the electron distribution perturbation in their amide bonds. The measurements were performed in dichloromethane (DCM). To confirm and rationalize the assignments, the spectra of the respective series of saturated Ac-Xaa-NHMe, recorded in DCM, and the spectra of these two series of unsaturated and saturated compounds, recorded in acetonitrile, were examined. To help interpret the spectroscopic results, the equilibrium geometrical parameters for some selected amides were used. These were optimized with ab initio methods in the 6-31G** basis set. Each of the dehydroamides studied adopted a C5 structure, which in Ac-delta Ala-NHMe is fully extended and accompanied by the strong C5 hydrogen bond. Interaction with the C alpha = C beta bond lessened the amidic resonance within each of the flanking amide groups. The N-terminal C = O bond was noticeably shorter, both amide bonds were longer than the corresponding bonds in the saturated entities and the N terminal amide system was distorted. Ac-delta Ala-NHMe constituted an exception. Its C-terminal amide bond was shorter than the standard one and both amide systems were prototypically planar. PMID- 9401919 TI - Reengineering a type II beta-turn as a potential helix nucleator. Part I. Crystal structure of Boc-Val-Pro-(D)Asp-Asp-Val-OMe monohydrate. AB - The crystal structure of Boc-Val1-Pro2-(D)Asp3-Asp4-Val5-OMe is described as a type II beta-turn reengineered into a potential helix nucleator. (D)Asp3 in the peptide is responsible for the configurationally guided LD chiral type II beta turn centered at Pro2-(D)Asp3, as well as the partially developed LL chiral type I beta-turn centered at Asp4-Val5 by acceptance of a conformation nucleating H bond from Val5NH to its carboxylic oxygen. PMID- 9401920 TI - Synthetic peptide vaccines: palmitoylation of peptide antigens by a thioester bond increases immunogenicity. AB - Synthetic peptides have frequently been used to immunize animals. However, peptides less than about 20 to 30 amino acids long are poor immunogens. In general, to increase its immunogenicity, the presentation of the peptide should be improved, and molecular weight needs to be increased. Many attempts have been made to couple peptide immunogens to different carrier proteins [e.g. keyhole limpet haemocyanin (KLH) or ovalbumin]. This leads to very complex structures, however. We used a controlled conjugation of a peptide to a single long-chain fatty acid like palmitic acid by a thioester or an amide bond. It was found that these S-palmitoylated peptides were much more immunogenic than N-palmitoylated peptides and at least similar to KLH-conjugated peptides with respect to appearance and magnitude of induced antibodies (canine parvovirus) or immunocastration effect (gonadotropin-releasing hormone). For chemical synthesis of thioesters, we established conditions for solution and solid-phase synthesis. In both phases, Cys(SBut) could only be deprotected efficiently using phosphines, and S-acylation was accomplished using standard coupling at pH 5. We speculate that, in vivo, the presence of an appropriate fatty acid chain, chemically linked through a labile thioester bond, greatly enhances immunogenicity, because it represents a favourable substrate for cleavage by cellular thioesterases in cells of the immune system. PMID- 9401921 TI - Common and differential recognition of structural features in synthetic peptides by the catalytic domain and the Src-homology 2 (SH2) domain of pp60c-src. AB - The relative efficiencies of the catalytic domain of the src-family kinase pp60c src in phosphorylating four peptide substrates including (i) src-optimal peptide (AEEEIYGEFEAKKKK), (ii) "-YEEI-peptide" (KKTHQEEEEPQYEEIPIYL), (iii) cdc2(6-20) (KVEKIGEGTYGVVYK), (iv) src-autophosphorylation site peptide (ADFGLARLIEDNEYTARG) and the relative efficiencies of its SH2 domain in binding the phosphorylated forms of these peptide substrates were compared. The results show that the src optimal peptide, "-YEEI-peptide," cdc2(6-20) peptide were phosphorylated by the catalytic domain with high efficiency and that the phosphorylated form of all three peptides could bind the SH2 domain of the kinase, confirming the hypothesis proposed by Songyang and co-workers that the catalytic domain of pp60c-src phosphorylates sites which are recognized by its own SH2 domain (Songyang et al. (1995) Nature 373, 536-539). The four peptides were phosphorylated by the kinase with relative efficiencies in the order of Src-optimal peptide > "-YEEI-peptide" > cdc2(6-20) >> src-autophosphorylation site peptide. However, the Tyr(P)-Src optimal peptide and [pY]15cdc2(6-20) bound to the SH2 domain of the kinase with an affinity at least an order of magnitude lower than that of the tight-binding peptide, "-pYEEI-peptide." Thus, our study suggests that the catalytic and SH2 domains of pp60c-src recognize overlapping but not identical determinants in the local structure around the tyrosine phosphorylation site of the substrate peptides. PMID- 9401922 TI - Aza-peptides. III. Experimental structural analysis of aza-alanine and aza asparagine-containing peptides. AB - To determine the structural perturbations induced by the C alpha H-->N alpha exchange in aza-peptides, we have examined by 1H NMR and IR spectroscopy various derivatives of the aza-analogues of alanine, aspartic acid and asparagine in different organic solvents with increasing polarity. Their general formulas are: R1-AzXaa-NR2R3, R1-Pro-AzXaa-NR2R3 and R1-AzXaa-Pro-NR2R3 (where AzXaa denotes the aza-analogue of the amino acid residue Xaa = Ala, Asp, Asn; R1 = Boc, Z; R2, R3 = H, Me, iPr). The aza-analogue of an amino acid residue appears to be a strong beta-turn-inducing motif, and the AzAsn carboxamide side-chain is capable of interacting, as a proton donor, with the preceding peptide carbonyl group. PMID- 9401923 TI - The nature of trypsin-pancreatic trypsin inhibitor binding: free energy calculation of Tyr39-->Phe39 mutation in trypsin. AB - The main goal of this work is the detailed study of the binding interactions in the trypsin-pancreatic trypsin inhibitor (PTI) complex and, here, we present how meaningful the Tyr39-Ile19 interaction is to the stability of that particular complex using free energy methods. This knowledge should be very important in the design of new inhibitors for trypsin and enzymes homologous to it. In particular, it could help to decide whether it is possible to produce selective inhibitors for these enzymes by appropriate mutations of residues in the contact region of PTI. PMID- 9401924 TI - Structure and supramolecular packing features of the dipeptide Arg-Val acetate. AB - The title compound crystallizes in the zwitterionic form. The crystal forms a supramolecular structure with the peptide molecules organized in head-to-tail columns in the b direction. The arginine side-chains and acetate ions interact with neighbor peptides in the c direction. Infinite hydrophobic columns are present in the a direction; they involve the valine side-chains, the acetate methyl groups and the methylene groups of the arginine side-chains. This three dimensional organization is similar to that found in Lys-Val hydrochloride. PMID- 9401925 TI - Technetium-99m labeled epidermal growth factor-tumor imaging in mice. AB - We have shown previously that the epidermal growth factor peptide (EGF) may be radiolabeled with 99mTc at room temperature and neutral pH by using the N hydroxysuccinimide ester of S-acetyl mercaptoacetyltriglycine (MAG3) as a bifunctional chelator. By a competition binding assay, we found that MAG3 conjugated EGF retained biological activity. Furthermore, the labeled peptide exhibited saturation binding to EGF receptor-positive tumor cell lines which could be inhibited by presaturation of the cells with unlabeled, native EGF. Biodistribution in normal mice at 3 h postadministration showed rapid clearance with minimal retention of the label in sampled organs. We have now investigated the tumor localization properties in mice of this labeled peptide. Nude mice implanted with the EGF receptor-positive tumors A431 and LS-174T were administered labeled EGF and a labeled control peptide (BPTI, aprotinin). Tumor uptake at 12 h postadministration was 0.44% injected dose/g for EGF/g vs. 0.09 for the control. Pretreatment of tumored mice with unlabeled EGF blocked about half the tumor uptake. Animals were also administered an anti-EGF receptor antibody labeled with 99mTc via MAG3. Relative to the antibody, tumor-to-muscle ratios were improved from 6 to 15 and tumor-to-blood ratios from 0.4 to 7 with EGF. These favorable results along with documented evidence of overexpression of the EGF receptor in many human tumors suggest that 99mTc-EGF should be considered further for tumor detection. PMID- 9401926 TI - Primary structures of proteins characterized by proteinase K digestion and matrix assisted laser desorption/ionization mass spectrometry. AB - To improve the sequence ions of a protein in matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), proteinase K was used to digest the protein followed by MALDI-MS characterization of the peptide fragments. The primary structures of three proteins, insulin B chain, cytochrome c and lysozyme, were determined by this method. A series of peptide fragments including those differentiated by one residue can be produced from the protein by using proteinase K digestion, thus providing support to the protein sequence. The peptide fragments liberated from proteinase K proteolysis of the insulin B chain allow the protein to be partially sequenced. Furthermore, some of the residues are double or triple checked by generating a variety of fragments. The same method was used to investigate cytochrome c and lysozyme denaturated in 3 M guanidine hydrochloride. The success of the method relies on the intrinsic properties of proteinase K and accurate determination of the peptide fragments by MALDI-MS. PMID- 9401928 TI - Periodontal conditions in 35-44-yr-old adults in France, 1993. AB - A national study was carried out in France in 1993 to assess the periodontal status of the population aged 35-44 yr. The study took part in the Second International Collaborative Study of Oral Health Outcomes developed and coordinated by the World Health Organization. The representative sample was composed of 1000 subjects. The Community Periodontal Index of Treatment Needs (CPITN) index was used. Gingivitis prevalence was high (80.4%) while 26.6% of dentate subjects had shallow pockets (4-5 mm). Deep pockets (> 6 mm) were rare (1.6%) concerning on average 0.1 sextant per subject; 87.5% of the 994 dentate adults needed periodontal treatment. Oral health education and scaling should reduce periodontal pathology in this population group. PMID- 9401927 TI - Interleukin-6 production in human fibroblasts derived from periodontal tissues is differentially regulated by cytokines and a glucocorticoid. AB - Interleukin-6 (IL-6) is thought to be a major mediator of the host's defense against infection, and it regulates immune responses in inflamed tissue. In this study, we investigated the regulation of IL-6 production in human gingival fibroblasts (HGF) and human periodontal ligament fibroblasts (HPLF). Pro inflammatory cytokines including interleukin (IL)-1 alpha, IL-1 beta and tumor necrosis factor (TNF)-alpha stimulated IL-6 production in HGF and HPLF in a time- and dose-dependent manner. This IL-1 alpha, IL-1 beta, or TNF-alpha-induced IL-6 production was enhanced, but the cAMP accumulation they induced was inhibited by the addition of indomethacin. This result suggests that endogenous prostaglandin E2 (PGE2) partially inhibits IL-1 or TNF-alpha-induced IL-6 production and that the enhancement of IL-6 production by IL-1 or TNF-alpha may not be caused through endogenous PGE2-induced cAMP-dependent pathway. Dexamethasone (DEX), a glucocorticoid which is a inhibitor of nuclear factor kappa B (NF-kappa B activation, markedly inhibited IL-1 (alpha or beta) or TNF-alpha-induced IL-6 production; so this production may be partially mediated through NF-kappa B. IL-1 (alpha or beta) and TNF-alpha enhanced IL-6 production synergistically. IL-6 production in HGF or HPLF stimulated with IL-1 beta was augmented by the addition of interferon (IFN)-gamma, but was slightly suppressed by the addition of IL-4. Endogenous IL-6 enhanced IL-1 (alpha or beta)-induced IL-6 production in the presence of IL-6 soluble receptor (IL-6sR). Accordingly, in inflamed periodontal tissues, gingival fibroblasts and periodontal ligament fibroblasts stimulated with pro-inflammatory cytokines such as IL-1 or TNF-alpha, may produce IL-6, and this production can be differentially modulated by endogenous PGE2, IL-6sR, T cell-derived cytokines such as IFN-gamma or IL-4, and glucocorticoids. PMID- 9401929 TI - Effect of the NSAID flurbiprofen on remodelling after periodontal surgery. AB - The aim of the present experiment was to assess the effect of the administration of the NSAID flurbiprofen (Froben) on tissue healing after periodontal surgery. Sites from patients with the same treatment modality (modified Widman flap) but receiving a placebo drug and sites within each patient not exposed to surgery served as controls. Nineteen patients suffering from moderate to severe periodontal disease were recruited and they signed informed consent forms. These patients required periodontal surgery as assessed at the periodontal re evaluation. The sites chosen for the study were all diagnosed with PPD > or = 5 mm and were bleeding on probing. During the healing phase 10 patients received 50 mg Froben 3 times per day for 30 d whereas 9 patients received a placebo drug. Two sites with PPD > or = 5 mm after initial therapy and bleeding on probing served as surgical sites, whereas 2 similar sites were not exposed to surgery. The study design was set up double-blind. The radiographic examination consisted of 2-4 standardized vertical bitewings obtained at the periodontal re-evaluation (BL) at 1, 3 and 6 months post-surgically for digital subtraction and computer assisted densitometric image analysis (CADIA). The regions of interest analysed were mesial or distal crestal sites. Minimal remodelling activity was observed radiographically after periodontal surgery in both patient groups. There were no statistically significant differences between the four groups of sites regarding the mean changes in density when analysing the pairs of radiographs 0-1, 0-3, 0-6 months. A frequency analysis was performed to list the number of sites with different ranges of density change. No differences in the distributions of the numbers of sites were observed when comparing the 4 site groups (Kolmogorov Smirnov, p > 0.05). A significant reduction of the probing pocket depth and a significant amount of clinical attachment gain was noted at the surgically treated sites irrespective of whether the patients had used flurbiprofen or placebo. Whereas the pathways leading to bone resorption in periodontally diseased sites have been shown, in other studies, to be influenced by NSAID, the results of the present study could not justify general administration of Froben for the purpose of reduction of bone resorption after periodontal surgical procedures in patients with adult periodontitis. PMID- 9401930 TI - Cystatin A in gingival crevicular fluid of periodontal patients. AB - Cystatins are physiological inhibitors of cysteine proteinases which are widely distributed in human tissues and fluids. In the present study we analysed both the cystatin activity and the different cystatin isoforms in gingival crevicular fluid and saliva samples of nine periodontitis patients. All crevicular fluid samples, which were collected with filter paper points, showed cystatin activity ranging from 7-67 units/mg protein. The mean cystatin activity (24 units/mg protein) was significantly lower (p < 0.05) than that of the saliva samples (mean 93 units/mg protein). The cystatin isoforms in the crevicular fluid were further characterized by immunoblotting with specific antibodies against cystatin C, S, SN and A. While they were clearly present in saliva, cystatin C, cystatin S and cystatin SN could not be detected in any of the crevicular fluid samples. Remarkably, cystatin A was found in all the crevicular fluids as well as in the saliva samples. It is concluded that the cystatin activity found in crevicular fluid is caused, at least partially, by cystatin A. Furthermore, the gingival crevicular fluid is not a major contributor of cystatin C, S and SN activity in saliva. PMID- 9401931 TI - Radiography of spontaneous periodontitis in dogs. AB - The pattern and distribution of periodontitis were investigated in 162 randomly selected dogs available for necropsy in veterinary practice. There were 82 males and 80 females of 50 different breeds (150 dogs were pure-bred and 12 were mongrels, aged between 7 months and 14 yr. Presence of periodontitis was determined by assessment of alveolar bone loss on radiographs of the skulls and jaws. Periodontitis occurred frequently with increasing age, although the prevalence varied markedly among and within different breeds. Of the breeds most represented in the sample, periodontitis was most frequently seen in poodles and dachshunds but was rarely recognized in German shepherd dogs. Regardless of age, the vast majority of the dogs displayed either one or both of two different radiographic patterns of alveolar bone loss. One pattern was characterized by slight, horizontal alveolar bone loss involving interradicular and interdental areas. The other pattern was one of predominantly crater-like, or narrow, vertical bone defects which, when advanced, often extended around a single root or tooth to surround the root apices. The two types of patterns did not seem to be breed-dependent. The posterior maxillary and mandibular premolars and molars were the most frequently affected teeth. Alveolar bone loss was most severe in the maxilla, while corresponding bone loss in the mandible was more often related to increasing age. PMID- 9401932 TI - An immunohistochemical study on the localization of Porphyromonas gingivalis, Campylobacter rectus and Actinomyces viscosus in human periodontal pockets. AB - The localization and distribution of Porphyromonas gingivalis, Campylobacter rectus and Actinomyces viscosus were studied in human periodontal pockets. After obtaining voluntary consent from 9 patients, 12 teeth and their surrounding periodontal tissue with advanced adult periodontitis were extracted carefully so as not to change the structure of the periodontal pockets. The specimens were processed into serial sections. One of the sections was stained with Brown & Brenn-modified Gram stain to observe the distribution of bacteria. The others were stained immunohistochemically by the Labelled Streptavidin Biotin method (LSAB method) using specific rabbit antibodies against selected bacteria. Some bacteria could be found within epithelial cells. P. gingivalis was found in 9/12 of the samples examined. Small aggregates of P. gingivalis were scattered in all parts of the periodontal pockets, and some of these aggregates could be seen in close contact with the epithelium. Conversely, C. rectus was observed in 5/12 of the samples examined and was predominantly located in the middle and deep pocket zones. C. rectus tended to form large clumps in both the tooth-attached and epithelium-associated plaque area. A. viscosus was observed in 7/12 of the samples examined and was localized predominantly in the tooth-attached plaque area, especially in the shallow and middle pocket zones. Although unexpected spills of unattached plaque from periodontal pockets was possible, immunohistochemical staining with species-specific antibodies was extremely sensitive and revealed the localization and the distribution of periodontal disease-associated bacteria in human periodontal pockets. PMID- 9401933 TI - Polyclonal B cell activators and in vitro induction of auto-antibody reactive with collagen. AB - Cells producing autoantibodies are known to be present in chronically inflamed periodontal tissues. In sites of chronic inflammation, polyclonal B cell activators (PBA) are known to exhibit adjuvant activity when combined with foreign antigens. These results prompted an examination of PBA in eliciting an antibody response to an autoantigen (i.e. collagen type I). Rat lymphocytes were stimulated with rat collagen (type I), microbial PBA (LPS) or the combination of LPS plus rat collagen in vitro. Anti-collagen antibody-forming cells (AFC) were enumerated using an ELISPOT assay. Collagen or LPS alone elicited few anti collagen AFC but the addition of LPS to collagen resulted in a substantial adjuvant effect and yielded maximal responses to collagen. Comparisons of anti collagen AFC from short-term immunized (2-6 wk after booster), non-immunized and long-term immunized (3-4 months after booster) animals were performed. It revealed that cells from recently immunized rats were significantly easier to activate than the other 2 groups. The adjuvant effect of microbial PBA may be important in anti-collagen antibody production and thus the localization of PBA in periodontal pockets may explain why anti-collagen AFC are restricted to the chronically inflamed periodontal tissues. PMID- 9401934 TI - Maintenance of new cementum formed during cyclosporin A administration after suspension of the treatment. AB - The aim of the present investigation was to examine if new cementum (NC) formed during cyclosporin A (CsA) administration was maintained after suspension of the treatment. Thirty mg/kg/d of CsA were given to 3 male Sprague-Dawley rats. Three control rats received oil-based vehicle solution. Nine wk later the drug and vehicle administration were stopped and the rats continued to be fed with the same standard laboratory diet and water ad libitum for 5 months. The rats were anaesthetized, the tissues fixed by intracardiac perfusion of fixative solution and the mandibles processed for Epon inclusion. Histological, histomorphometric and ultrastructural analysis revealed that (a) NC covered extensive areas of the root surfaces; its structural characteristics were identical to those observed in the rats killed during CsA administration. (b) collagen fibres of the adjacent connective tissue were functionally inserted into the NC. (c) In the presence of cervical NC spurs the extent of the apical downgrowth of the junctional epithelium, measured parallel to the cemento-dentinal junction, was decreased (up to 64%) compared to the one occurring in areas devoid of NC deposits. These results suggest that (a) NC deposition and its functional relations with the adjacent connective tissue are not reversible after cessation of CsA treatment and (b) in the presence of cervical NC spurs the amount of connective tissue attachment on the root surfaces is increased. PMID- 9401935 TI - Attachment loss with postmenopausal age and smoking. AB - To determine whether postmenopausal bone loss and factors associated with osteoporosis affect tooth retention, we examined vertebral and proximal femoral (postcranial) bone mineral density in relation to tooth loss and attachment loss in a cross-sectional study of 135 postmenopausal women (age range 41-70 yr). Women had at least 10 teeth and no evidence of moderate or severe periodontal disease. Full-mouth attachment loss measurements were made using a pressure sensitive probe, and bone density was determined by dual-energy X-ray absorptiometry. Attachment loss was correlated with tooth loss (number of remaining teeth, radiologically determined), but not with vertebral or proximal femur bone density. Multivariate analysis showed current smoking (p = 0.01), years since menopause (p = 0.02) and the interaction of age and current smoking (p < 0.01), to be statistically significant predictors of attachment loss in our study population. PMID- 9401936 TI - A hydrogel based on a polyaspartamide: characterization and evaluation of in-vivo biocompatibility and drug release in the rat. AB - This paper deals with the characterization of a new microparticulate hydrogel obtained by gamma irradiation of alpha, beta-poly[N-(2-hydroxyethyl)-DL aspartamide] (PHEA). When enzymatic digestion of PHEA hydrogel was evaluated using various concentrations of pepsin and alpha-chymotrypsin no degradation occurred within 24 h. In-vivo studies showed that this new material is biocompatible after oral administration to rats. PHEA hydrogel was also studied as a system for delivery of diflunisal, an anti-inflammatory drug. In-vitro release studies in simulated gastrointestinal juice (pH 1 or 6.8) showed that most of the drug was released at pH 6.8. In-vivo studies indicated that diflunisal-loaded PHEA microparticles significantly improved the gastric tolerance and oral bioavailability of the drug in comparison with free diflunisal. These results suggest the potential application of PHEA hydrogel as a new delivery system for the oral administration of anti-inflammatory drugs. PMID- 9401937 TI - Effects of different absorption enhancers on the permeation of ebiratide, an ACTH analogue, across intestinal membranes. AB - The permeation of ebiratide (H-Met(O2)-Glu-His-Phe-D-Lys-Phe-NH(CH2)8NH2), a novel ACTH analogue, across the intestinal mucosae has been examined by use of isolated intestinal membranes from rats in a modified Ussing chamber. Regional differences were observed in the permeation of ebiratide across intestinal membranes; the order of membrane permeability was jejunum > ileum > duodenum > colon. Overall, the permeation of ebiratide was relatively poor. The effects of various absorption enhancers were examined to increase the intestinal permeability to ebiratide. Sodium glycocholate and sodium caprate had no significant enhancing effect on the permeability of the jejunal membrane, but significantly enhanced the permeation of ebiratide through the colonic membrane. On the other hand, N-dodecyl-beta-D-maltopyramoside (LM) significantly enhanced the permeation of ebiratide through both jejunal and colonic membranes. In general, the absorption-enhancing effects of these agents were more predominant in the colon than in the jejunum. Membrane damage by the absorption enhancers was evaluated by measuring the amount of protein released from the intestinal membrane. It was found that all the absorption enhancers slightly increased the amount of protein released, but that the amounts of protein released in the presence of these enhancers were much less than in the presence of ethylenediaminetetraacetic acid (EDTA), used as a positive control. These findings suggest that the absorption enhancers, especially LM might be useful adjuvants for improving the intestinal absorption of peptide and protein drugs, including ebiratide. PMID- 9401938 TI - Airway reactivity to inhaled spasmogens 18-24 h after antigen-challenge in sensitized anaesthetized guinea-pigs. AB - The anaesthetized allergic guinea-pig was used to assess changes in airway reactivity to four different inhaled spasmogens: methacholine, 5 hydroxytryptamine (5-HT), histamine and the thromboxane A2 mimetic, 9,11-dideoxy 9 alpha,11 alpha-methano-epoxy-PGF2 alpha (U-46619). Reactivity was determined 18 to 24 h after challenge of ovalbumin-sensitized guinea-pigs with inhaled ovalbumin. This time coincides with the appearance of a late-phase bronchoconstriction in these animals. Sensitivity to the spasmogen was assessed from the concentration-response curve for the increase in pulmonary inflation pressure (PIP) in ovalbumin- and saline-challenged sensitized animals. When methacholine, 5-HT or histamine were the spasmogens there was no hyper reactivity. The geometric mean EC50 values (i.e. the concentrations inducing half the maximum effect) obtained from the dose-response curves for methacholine (73 (42-129) and 94 (66-134) micrograms mL-1), 5-HT (1.5 (0.81-3.03) and 1.1 (0.51 2.24 micrograms mL-1) and histamine (39 (21-75) and 72 (32-162) micrograms mL-1) did not differ significantly (P > 0.05) between saline- and ovalbumin-challenged animals, respectively. However, when U-46619 was the spasmogen, ovalbumin-induced airway hyper-reactivity was observed as a leftwards shift of the concentration response curve and the EC50 value for ovalbumin-challenged animals (8.1 (5.1-13) ng mL-1) was significantly (P < 0.05) less than the value for control animals (39 (21-75) ng mL-1). Our findings suggest that airway hyper-reactivity is not 'non specific', but instead depends on the chosen spasmogen. The absence of hyper reactivity with certain spasmogens was not a result of poor delivery, because all spasmogens caused a bronchoconstriction by the inhaled route. It was also not associated with the model because ozone has been shown to induce hyper-reactivity to inhaled methacholine and 5-HT. Because airway hyper-reactivity to both inhaled histamine and agonists at muscarinic receptors is regularly seen in man, the anaesthetized guinea-pig might not be the ideal model for assessing airway hyper reactivity after antigen challenge and its modification by anti-asthma drugs. PMID- 9401940 TI - Comparison of the effects of cimetidine and ranitidine on the pharmacokinetics of disopyramide in man. AB - The widely prescribed antiulcer agents cimetidine and ranitidine have the potential to affect the absorption, metabolism or renal excretion of disopyramide. This study investigated the effect of a single oral dose of cimetidine or ranitidine on the pharmacokinetics of disopyramide and mono-N dealkyldisopyramide in six healthy volunteers. The treatment was conducted in a randomized cross-over design. Serum levels and urinary recoveries of disopyramide and mono-N-dealkyldisopyramide were assayed by HPLC. Cimetidine significantly elevated the maximum plasma concentration of disopyramide, the area under the plasma concentration-time curve and the total amount of disopyramide excreted unchanged in the urine, but the serum profile of mono-N-dealkyldisopyramide was not significantly affected. The effects of ranitidine on the pharmacokinetics of disopyramide and mono-N-dealkyldisopyramide were not significant. The interaction between cimetidine and disopyramide occurred mainly at the site of absorption. The results indicate that cimetidine, but not ranitidine, significantly increased the absorption of orally administered disopyramide. PMID- 9401939 TI - Intestinal absorption of stable cyclic glycylphenylalanine: comparison with the linear form. AB - The absorption, especially the stability and transportability, of the cyclic peptide cyclic glycylphenylalanine (cyclo(Gly-Phe)) and the linear peptides glycylphenylalanine, glycyl-D-phenylalanine and phenylalanylglycine have been studied in rat small intestine. Linear peptides were degraded on the mucosal side and only glycyl-D-phenylalanine appeared on the serosal side. However, cyclo(Gly Phe) was stable on the mucosal side and appeared on the serosal side. Furthermore, the absorption clearance of cyclo(Gly-Phe) was higher than that of glycyl-D-phenylalanine. In the presence of the peptidase inhibitor bestatin, the degradation of linear peptides was reduced and linear peptides appeared on the serosal side, but only phenylalanylglycine, which is transported by the oligopeptide transporter, was absorbed faster than cyclo(Gly-Phe). The absorption clearance of cyclo(Gly-Phe) was reduced as its concentration was increased from 125 microM to 500 microM. Furthermore, the absorption clearance of cyclo(Gly-Phe) at 125 microM was reduced at 4 degrees C or in the presence of glycylsarcosine and cephalexin, which are transported by the oligopeptide transporter. These results indicated that cyclo(Gly-Phe) was stable enough to be absorbed and was transported in part by the oligopeptide transporter rather than completely by passive diffusion. PMID- 9401941 TI - Pharmacokinetics of N4-octadecyl-1-beta-D-arabinofuranosylcytosine in plasma and whole blood after intravenous and oral administration to mice. AB - N4-octadecyl-1-beta-D-arabinofuranosylcytosine (NOAC) is a new cytotoxic derivative of cytosine arabinoside with improved cytotoxic activity and stability against deamination. Its pharmacokinetics were studied in mice. The drug was administered intravenously and orally to ICR mice to assess its pharmacokinetic parameters in plasma and whole blood. The lipophilic drug was administered in small unilamellar liposomes 100-400 nm in diameter. The concentrations of NOAC in plasma and erythrocytes were determined by high-performance liquid chromatography (HPLC). When given orally a rather low amount of the delivered NOAC was absorbed as the unchanged drug, resulting in a bioavailability of 1.1% from the plasma and 12.9% from whole blood. As shown elsewhere, the amount of drug absorbed is sufficient to provide excellent cytotoxic activity in the L1210 leukemia and in human xenograft models after oral administration. The mean residence time of NOAC after intravenous administration was 3.5 h in plasma and 6 h in whole blood giving NOAC a terminal half-life in blood substantially longer than that of cytosine arabinoside. After oral administration the mean residence time was 18 h in plasma and whole blood. In summary, NOAC has a prolonged half-life after intravenous administration compared with cytosine arabinoside. The distribution of NOAC in blood is highly dependent on its mode of administration. PMID- 9401943 TI - Stereoselective passage of mefloquine through the blood-brain barrier in the rat. AB - The pharmacokinetics of the enantiomers of mefloquine were studied in the rat after administration of a racemic mixture and of the separate enantiomers (+) mefloquine and (-)-mefloquine. When 50 mg kg-1 racemic mixture was administered orally for 22 days, plasma concentrations of the (+) enantiomer were 2-3 times higher than those of the (-) enantiomer whereas the opposite was true in every part of the brain (cerebellum, cortex, hippocampus, hypothalamus and striatum). Different concentrations of mefloquine were found in the different regions of the brain; the lowest concentrations of (+/-)-mefloquine (27.0 nmol g-1) were in the cerebellum and the highest (110.0 nmol g-1) in the hippocampus. The main metabolite, carboxymefloquine, was detected in plasma but not in the brain. The results indicate the mefloquine crosses the blood-brain barrier stereoselectively. PMID- 9401942 TI - Pharmacokinetics of intragastrically administered digoxin in rabbits with experimental bile duct obstruction. AB - A change in the functioning of the liver as a result of experimental cholestasis could result in a change in the biotransformation of drugs. The aim of this study was to evaluate the effect of extrahepatic cholestasis on the pharmacokinetics of digoxin. The investigation was performed on male rabbits randomly divided into two groups: sham-operated and animals with bile-duct ligation. Digoxin (0.02 mg kg-1) was administered intragastrically as a single dose. Biomedical and anatomo pathological tests and pharmacokinetic assays were performed before the operation and on the 6th day after surgery. A significant increase in area under the serum concentration-time curve and in mean residence time, a decrease in total body clearance, a reduction in the volume of distribution and increases in maximum concentration and the time to reach maximum concentration were observed in animals with the bile-duct ligation. These results suggest reduced elimination of digoxin in animals with obstructive cholestasis. PMID- 9401944 TI - Affinity of the miotic drug, dapiprazole, at alpha 1-adrenoceptor subtypes A, B and D. AB - The functional affinities of the alpha 1-adrenoceptor antagonist, dapiprazole, currently being used to reverse diagnostic pupillary dilation, were determined at subtype A in rat vas deferens, at subtype B in guinea-pig spleen and at subtype D in rat aorta and compared with various alpha 1-adrenoceptor subtype discriminating antagonists. Dapiprazole had relatively high affinity both at rat vas deferens alpha 1A-adrenoceptors (pA2 = 7.93) and at rat aortic alpha 1D adrenoceptors (pA2 = 8.26), whereas its affinity at guinea-pig splenic alpha 1B adrenoceptors (pA2 = 7.13) was lower. The reference antagonists, 5-methylurapidil and the 5-methylurapidil/flesinoxan hybrid, B8805-033 ((+/-)- 1,3,5-trimethyl 6[[3[4(2(2,3-dihydro-2-hydroxymethyl)-1,4-benzodioxin -5-yl)-1 piperazinyl]propyl]-amino]2,4(1H,3H)-pyrimidinedione), were 40- and 1500-fold selective for the A subtype, whereas spiperone and BMY 7378 (8-[2-[4-(2 methoxyphenyl)-1- piperazinyl]ethyl]-8-azaspiro[4,5]decane-7,9-dione diHCI) were confirmed as selective for the B and D subtypes of alpha 1-adrenoceptors, respectively. Thus, in functional experiments dapiprazole seems to be moderately selective (approximately 10-fold) for the A and D over the B subtype of alpha 1 adrenoceptors; the possible therapeutic consequence of this is discussed. PMID- 9401945 TI - Cold-storage of rabbit thoracic aorta in University of Wisconsin solution reduces endothelium-independent vasodilation. AB - Optimum preservation conditions for storage of donor livers and blood vessels are essential for successful transplantation. The blood vessels are used as vascular conduits to facilitate anastomosis of the liver to the recipient's systemic vasculature. Failure of some transplants has been ascribed to thrombosis of these vascular conduits possibly because of alterations in vascular reactivity owing to inadequate storage techniques. To restrict data variability previously associated with studies using a heterogeneous sample of vessels from man, this study investigated changes in vascular reactivity in segments of rabbit thoracic aorta from male, age-matched, New Zealand White rabbits stored at 4 degrees C in either University of Wisconsin solution (UW; Du Pont Pharmaceuticals, UK) or Krebs Bulbring buffer (KB). Percent vasodilation to acetylcholine remained significantly greater in UW than in KB at -log (M) concentrations of 7.0 (UW = 47.05 +/- 4.26 compared with KB = 13.20 +/- 7.20%; P < 0.001), 6.5 (UW = 66.82 +/ 4.83 compared with KB = 26.60 +/- 9.48%; P < 0.01), and 6.0 (UW = 83.68 +/- 5.26 compared with KB = 31.20 +/- 9.83%; P < 0.001). This was not significantly different to relaxation in unstored arteries and suggested improved endothelial function and structure, confirmed by electron microscopy. Percent vasodilation to sodium nitroprusside was significantly lower in UW than in unstored (D0) arteries at -log (M) concentrations of 7.5 (D0 = 28.27 +/- 4.02 compared with UW = 15.21 +/- 1.82%; P < 0.01), 7.3 (D0 = 52.58 +/- 5.05 compared with UW = 29.23 +/- 1.94%; P < 0.01), 7.0 (D0 = 69.70 +/- 4.85 compared with UW = 49.72 +/- 2.49%; P < 0.05), and 6.4 (D0 = 93.16 +/- 2.93 compared with UW = 71.29 +/- 5.20%; P < 0.05). Percent vasodilation was also lower in UW- compared with KB-stored arteries at -log (M) sodium nitroprusside concentrations of 7.0 (UW = 49.72 +/- 2.49 compared with KB = 64.11 +/- 5.03%; P < 0.05) and 6.4 (UW = 71.29 +/- 5.20 compared with KB = 96.91 +/- 5.96; P < 0.05). Electron microscopy confirmed that this was not a result of degradation of smooth muscle structure. The nitric oxide synthase inhibitor L-NG-nitro-L-arginine methyl ester (100 microM) did not significantly modulate sodium nitroprusside-induced vasodilation in unstored arteries, when endothelial function was maximum, or in UW-stored arteries, suggesting that the reduced responses in UW-stored arteries were not because of increased synthesis of nitric oxide. This reduced relaxation to sodium nitroprusside was therefore nitric oxide-independent and not a result of competition between sodium nitroprusside and endothelial 'nitric oxide donation' for cGMP. In summary, cold-storage preservation with UW reduced endothelium independent vascular relaxation by mechanisms other than competition with NO; this requires further evaluation. PMID- 9401946 TI - Twenty-four-hour variations in the effect of nitrodilators in rat aorta: lack of influence of the endothelium. AB - Time-dependent variations of the vasodilator effects of sodium nitroprusside and glyceryl trinitrate on isolated smooth muscle have been studied on rings of rat thoracic aorta, both endothelium-intact and endothelium-denuded. For most of the concentrations of sodium nitroprusside used the induced relaxations were significantly dependent on the time the tissues were obtained. However, significant temporal differences were obtained for glyceryl trinitrate-induced relaxations at lower concentrations only for both endothelium-intact and endothelium-denuded preparations. EC50 values of sodium nitroprusside and glyceryl trinitrate (i.e. the concentrations inducing half the maximum response) were also significantly different and they had quite similar rhythmic features both in endothelium-intact and in endothelium-denuded preparations. These results clearly show that the in-vitro sensitivity of rat thoracic aorta to nitrodilator agents varies over a 24-h period and thus depends on when the animals were killed; the presence of endothelium does not change the rhythm of nitrodilator activity. These variations might be a result of circadian rhythm in the guanylate cyclase-cGMP system which mediates responses to nitrodilator agents. PMID- 9401947 TI - Effect of ganglionic blocking compounds on in-vivo fluid secretion in the rat small intestine. AB - It is well-known that enteric, secreto-motor nerves mediate cholera toxin-induced fluid secretion in the rat small intestine. This notion is, in part, derived from experiments on anaesthetized animals in which the response to cholera toxin was antagonized by the ganglionic nicotinic receptor antagonist, hexamethonium. In the current study, such anti-secretory action of ganglionic blocking compounds was analysed in an experiment designed to minimize any possible negative effect of general anaesthesia on intestinal secretion. Rats were anaesthetized with ether for 5-10 min, during which time a jejunal loop (10-12 cm) was constructed. The loop was challenged with one of the secretagogues, cholera toxin, prostaglandin E1 (PGE1) or okadaic acid. Saline (control) or either of the ganglionic blockers, hexamethonium and chlorisondamine, was administered intravenously. The rats were killed 5 h (cholera toxin) or 1.5 h (PGE1 and okadaic acid) after challenge, and the amount of fluid accumulated in the loops was determined. Cholera toxin-induced secretion was unchanged by hexamethonium but reduced by approximately 80% by chlorisondamine. The difference in effect between the two blockers might relate to the duration of ganglionic blockade. Chlorisondamine blocked secretion induced by either PGE1 or okadaic acid by approximately 60%. It is suggested that the anti-secretory effect of ganglionic blocking compounds might be a result of blockade of secreto-motor nerves but other mechanisms, for example interference with haemodynamic factors, cannot be ruled out. PMID- 9401948 TI - Several receptor subtypes contribute to 5-hydroxytryptamine-induced secretion by rat ileum in-vitro. AB - The receptors contributing to 5-hydroxytryptamine (5-HT)-induced secretion by rat ileum were investigated in-vitro using selective agonists and antagonists. 5-HT induced a dose-dependent increase in the short-circuit current (SCC) generated by both intact and stripped sheets of rat ileum. 1-Phenylbiguanide, a selective 5 HT3 agonist, and 5-methoxytryptamine, an agonist that lacks affinity for 5-HT3 receptors, also increased the SCC. In intact sheets 5-HT was more effective than either 1-phenylbiguanide or 5-methoxytryptamine, whereas in stripped sheets 5-HT and 5-methoxytryptamine were equipotent, with 1-phenylbiguanide having little effect. Tetrodotoxin abolished the response of intact sheets to 1-phenylbiguanide but only reduced the responses to 5-HT and 5-methoxytryptamine by 57% and 54%, respectively. This inhibition was reduced to 25% in stripped sheets. The 5-HT3 antagonist granisetron abolished the response to 1-phenylbiguanide, but did not alter the effects of 5-HT. Ketanserin, a 5-HT2 antagonist, had a small effect on the actions of 5-methoxytryptamine in intact, but not stripped, sheets and no effect on the response to 5-HT in either preparation. Tropisetron, a 5-HT3 and 5 HT4 antagonist, inhibited the response to 5-methoxytryptamine, but had less effect on the response to 5-HT. Desensitization to 1-phenylbiguanide inhibited the response to 5-HT in intact, but not stripped sheets, whereas desensitization to 5-methoxytryptamine abolished the 5-HT response in stripped sheets, but induced only 42% inhibition in intact sheets. Previous exposure to a combination of both 1-phenylbiguanide and 5-methoxytryptamine abolished the 5-HT-induced increase in SCC in both preparations. The findings suggest that 5-HT-induced ileal secretion involves more than one 5-HT receptor subtype and that both neural and non-neural mechanisms contribute to the response. PMID- 9401949 TI - Stereoselectivity of butylidenephthalide on voltage-dependent calcium channels in guinea-pig isolated ileum. AB - Two geometric isomers, the Z- and the E- forms, can be separated from synthetic mixtures of butylidenephthalide (Bdph). Z-Bdph (50-100 microM) non-competitively inhibited Ca(2+)-induced contractions in depolarized (K+, 60 mM) guinea-pig ileum longitudinal smooth muscle, with a pD2' value of 3.88 +/- 0.20 (n = 5). However, E-Bdph (20-100 microM) competitively inhibited these contractions with a pA2 value of 4.56 +/- 0.18 (n = 5) which was significantly (P < 0.05) greater than the pD2' value of Z-Bdph. In contrast, the two isomers had no stereoselective inhibitory action on Ca2+ influx through pre- or post-junctional membranes of cholinergic nerve endings from which the transmitter acetylcholine is released or on Ca2+ release from intracellular stores. Therefore, the trans-Z and cis-E forms of Bdph might have geometric stereoselectivity for voltage-dependent calcium channels (VDC) in guinea-pig longitudinal smooth muscle. Both isomers might inhibit more selectively the contractile twitch responses evoked by electrical stimulation than by cumulative acetylcholine- or carbachol-induced transient contractions in guinea-pig ileum longitudinal smooth muscle. PMID- 9401950 TI - Comparison of the intestinal secretory response to 5-hydroxytryptamine in the rat jejunum and ileum in-vitro. AB - A secretory response to 5-hydroxytryptamine (5-HT) is observed throughout the intestinal tract; this investigation has compared the nature of this response in the jejunum and ileum of the rat in-vitro. Different basal electrical activity was observed for jejunal and ileal sheets of rat small intestine. In both intact and stripped preparations the basal short-circuit current (SCC) was greater and the tissue resistance lower in the jejunum than in the ileum. 5-HT caused concentration-dependent increases in SCC in intact and stripped preparations of both regions. EC50 values were similar in the jejunum and ileum, stripped sheets from both regions showing greater sensitivity. In the ileum the maximum increase in SCC induced by 5-HT was similar in intact and stripped sheets, but in the jejunum the response was greater in intact preparations. The jejunal response to 5-HT was reduced in the absence of bicarbonate but unaffected by lack of chloride, whereas the ileal response was inhibited by removal of chloride but unaltered in bicarbonate-free conditions. In intact sheets the tetrodotoxin sensitive neural component was greater in the jejunum. In stripped sheets a neural component could still be detected in the ileum, but not in the jejunum. There are, therefore, fundamental differences in the way in which the jejunum and ileum respond to 5-HT stimulation--the jejunal response is primarily a result of stimulation of bicarbonate secretion whereas chloride secretion predominates in the ileum. The myenteric plexus appears to play a more prominent role in the jejunum; in the ileum other neural elements also contribute to the response. PMID- 9401951 TI - Screening of hepatoprotective plant components using a HepG2 cell cytotoxicity assay. AB - Identification of the active components of plants with hepatoprotective properties requires screening of large numbers of samples during fractionation and purification. A screening assay has been developed based on protection of human liver-derived HepG2 cells against toxic damage. Various hepatotoxins were incubated with HepG2 cells in 96-well microtitre plates (30,000 cells well-1) for 1 h and viability was determined by metabolism of the tetrazolium dye 3-(4,5 dimethylthiazol-2-yl)-5-(3-carboxymethoxy phenyl)-2-(4-sulphophenyl)-2H tetrazolium (MTS). Bromobenzene (10 mM) and 2,6-dimethyl-N-acetyl-p-quinoneimine (2,6-diMeNAPQI, 200 mM) had greater toxic effects than tert-butyl hydroperoxide (1.8 mM) or galactosamine (10 mM), reducing mean viability to 44.6 +/- 1.2% (s.e.m.) and 56.1 +/- 2.1% of control, respectively. Protection against toxic damage by these agents was tested using a crude extract of a known hepatoprotective Sri Lankan plant, Osbeckia aspera, and two pure established hepatoprotective plant compounds, (+)-catechin and silymarin (1 mg mL-1). Viability was significantly improved by Osbeckia (by 37.7 +/- 2.4%, P < 0.05, and 36.5 +/- 2.1%, P < 0.05, for bromobenzene and 2,6-diMeNAPQI toxicity, respectively). Comparable values for (+)-catechin were 68.6 +/- 2.9% and 63.5 +/- 1.1%, and for silymarin 24.9 +/- 1.4% and 25.0 +/- 1.6%. This rapid and reproducible assay should prove useful for the isolation and identification of active hepatoprotective compounds in crude plant extracts. PMID- 9401952 TI - Cisplatin-induced changes in adenine nucleotides in rat kidney slices: amelioration by tiopronin and procaine. AB - The adenine nucleotides (ATP, ADP and AMP) in rat renal cortical slices exposed in-vitro to cisplatin, an anticancer drug, were determined by HPLC. Cisplatin had no effect on total adenine nucleotides in the slices but caused a time- and concentration-dependent decrease in ATP levels with a concomitant increase in ADP and AMP levels. The decrease in ATP and increases in ADP and AMP concentrations became statistically significant after incubation with cisplatin (2 mM) for 90 min or after cisplatin (1 mM) for 120 min. Both tiopronin, a sulphydryl containing drug, and procaine, an antioxidant, protected against cisplatin induced changes in the adenine nucleotides. The results indicate a cisplatin induced defect in cellular energetics that occurs at a relatively late stage in the process of toxicity to the slices in this in-vitro model. Cisplatin-induced depletion of ATP in the slices might result from an increase in catabolism of ATP to ADP and AMP. Maintenance of the normal concentration of ATP in the slices might be involved in the protection afforded by tiopronin and procaine against cisplatin-induced nephrotoxicity. PMID- 9401953 TI - Application of muscle microdialysis to evaluate the concentrations of the fluoroquinolones pazufloxacin and ofloxacin in the tissue interstitial fluids of rats. AB - Muscle microdialysis has been used to determine the unbound concentrations of the fluoroquinolones, pazufloxacin and ofloxacin, in tissue interstitial fluids (Cisf,u) of rats under steady state conditions. Cisf,u was estimated from the concentration in dialysate and the in-vitro permeability rate constant by the extrapolation method based on the clearance concept. Paper-disks were inserted under the abdominal skin of rats, and the drug concentrations in the fluids penetrating into the disks (Cdisk) were measured and compared with Cisf,u. The Cisf,u of pazufloxacin and ofloxacin in muscle were close to their unbound concentrations in the venous plasma; these were 75.3% and 77.1%, respectively, of the total concentrations in plasma at the steady state. The Cdisk of pazufloxacin and ofloxacin were also close to their Cisf,u. These results indicate that the unbound concentrations of the fluoroquinolones in the tissue interstitial fluids were the same as those in the venous plasma. The disk insertion technique seems to be useful for evaluating drug concentrations in tissue interstitial fluid. PMID- 9401954 TI - Isoquinoline derivatives isolated from the fruit of Annona muricata as 5-HTergic 5-HT1A receptor agonists in rats: unexploited antidepressive (lead) products. AB - The fruit and the leaves of Annona muricata (Annonaceae) are used in traditional medicine for their tranquillizing and sedative properties. Extracts of the plant have been shown to inhibit binding of [3H]rauwolscine to 5-HTergic 5-HT1A receptors in calf hippocampus, and three alkaloids, annonaine (1), nornuciferine (2) and asimilobine (3), isolated from the fruit have been shown to have IC50 values of 3 microM, 9 microM and 5 microM, respectively, although in ligand binding studies it was not possible to determine whether interaction of these ligands with the receptor was agonistic or antagonistic. This paper presents the results of functional assays of the alkaloids. The inhibition of cAMP accumulation was tested in NIH-3T3 cells stably transfected with the 5-HT1A receptor from man. None of the alkaloids showed antagonistic properties towards the 5-HT1A receptors because in the antagonistic tests no influence on the forskolin-stimulated increase of cAMP level was detected. Full agonistic properties were measured for all three compounds; the inhibition constants (Ki) for 1, 2 and 3 were < 10 microM. Inhibition of the binding of the radioligand to the 5-HT1A receptor was observed in every ligand-binding assay performed with the alkaloids; the Ki values for 1, 2 and 3 were in the microM range. These results imply that the fruit of Annona muricata possesses anti-depressive effects, possibly induced by compounds 1, 2 and 3, and that in the past potent leads for the development of anti-depressive therapeutics have not been used. PMID- 9401955 TI - Cirsimarin and cirsimaritin, flavonoids of Microtea debilis (Phytolaccaceae) with adenosine antagonistic properties in rats: leads for new therapeutics in acute renal failure. AB - In traditional medicine Microtea debilis is used against proteinuria. In ligand binding studies extracts of Microtea debilis have been shown to inhibit the binding of [3H]1,3-dipropyl-8-cyclopentylxanthine ([3H]DPCPX) to adenosine-A1 receptors in rat forebrain membranes. Subsequently, cirsimarin, a flavonoid, was isolated as the active component and was shown to function as adenosine antagonist at the adenosine-A1 receptor in-vitro. In this study we have investigated the adenosine-A2 receptor activity of cirsimarin the in-vivo inhibition of the effects of adenosine by cirsimarin in rats, the absorption of cirsimarin and the inhibition of the binding of [3H]DPCPX to the adenosine-A1 receptor by urine samples obtained after oral administration of crude extract of Microtea debilis, cirsimarin or cirsimaritin to rats. Cirsimarin inhibited the binding of [3H]5'-N-ethylcarboxamidoadenosine ([3H]NECA) to adenosine-A2 receptors in rat striatum with an inhibition constant, Ki, of 6.5 +/- 0.3 microM. The decrease of heart rate and blood pressure induced by adenosine was significantly inhibited by cirsimarin. After oral administration of 8 and 80 mg kg-1 cirsimarin, the compound could not be detected in either plasma or urine, but the presence of cirsimaritin was established. By use of beta-glucuronidase, glucuronides of cirsimaritin were also detected in the urine. The concentrations of cirsimaritin in the plasma were 0.126 +/- 0.04, 0.138 +/- 0.015, and 0.120 +/- 0.022 microM, respectively, 2, 5 and 12 h after administration of 8 mg kg-1 cirsimarin. The concentrations of cirsimaritin in the urine at the same times after administration of the same dose were 2.05 +/- 1.86, 5.05 +/- 2.6 and 2.06 +/- 0.09 microM, respectively. The inhibition of the binding of [3H]DPCPX to the adenosine-A1 receptor by urine samples collected 2, 5 and 12 h after oral administration of 8 mg kg-1 cirsimarin or a crude extract of Microtea debilis containing approximately 8 mg kg-1 cirsimarin and 2.8 mg kg-1 cirsimaritin, or 6.8 mg kg-1 cirsimaritin, was not significantly different from that of urine samples collected from untreated rats, in contrast with urine samples collected 1 and 2 days after oral administration of 80 mg kg-1 cirsimarin. Approximately 3% of the cirsimarin was excreted in the urine as cirsimaritin. The results indicate that in the kidney and urinary tract the concentrations of cirsimaritin produced after ingestion of more than 8 mg kg-1 cirsimarin can be high enough to inhibit the interaction of adenosine with its receptors; this might explain the effectiveness of Microtea debilis preparations against proteinuria in traditional medicine. PMID- 9401956 TI - Evaluation of the long-term effects of oleum origani on the toxicity induced by administration of streptozotocin in rats. AB - Oleum origani, the essential oil of Origanum onites L., is a traditional plant material used in Turkey for the treatment of several diseases, including diabetes mellitus. This study has evaluated the effect of oleum origani on streptozotocin induced tissue injury and haematological changes. The effect of oleum origani on glycaemia was also studied. Long-term administration of oleum origani resulted in significant improvement of tissue injury induced by streptozotocin treatment. No effect on blood glucose levels was detected. In addition, any visible toxicity or disturbance of haematological parameters and tissue structure attributable to the long-term use of oleum origani were not established in normal rats. The data indicate that long-term use of oleum origani might be effective in preventing or at least in retarding the development of some complications of diabetes mellitus. Further investigation is required to determine the underlying mechanism(s) of the protective effect against tissue injury induced by streptozotocin-treatment of rats. PMID- 9401957 TI - The development of nao li shen and its clinical application. AB - The traditional Chinese medicine Nao Li Shen (containing ginseng, gastrodia tuber, chuanxiong rhizome and red sage root) is used in craniocerebral injury, cervical spondylosis and cerebrovascular diseases. The preparation, as an orally administered liquid, was tested in Mongolian gerbils and shown to increase tolerance to ischaemia and anoxia. Clinical use of the preparation resulted in improvement in 96% of 202 patients, as judged by right cerebral blood flow, TCD and CT examination. We conclude that Nao Li Shen has a positive curative effect upon craniocerebral injury and sequelae of cerebrovascular diseases. PMID- 9401958 TI - Detection of a black deposit in intravenous fat emulsions. PMID- 9401959 TI - Kinetic analysis of the phosphorylation-dependent interactions of synapsin I with rat brain synaptic vesicles. AB - 1. Synapsin I, a major synaptic vesicle (SV)-associated phosphoprotein, is involved in the regulation of neurotransmitter release and synapse formation. By binding to both phospholipid and protein components of SV with high affinity and in a phosphorylation-dependent fashion, synapsin I is believed to cluster SV and to attach them to the actin-based cytoskeleton of the nerve terminal. 2. In the present study we have investigated the kinetic aspects of synapsin I-SV interactions and the mechanisms of their modulation by ionic strength and site specific phosphorylation, using fluorescence resonance energy transfer between suitable fluorophores linked to synapsin I and to the membrane bilayer. 3. The binding of synapsin I to the phospholipid and protein components of SV has fast kinetics: mean time constants ranged between 1 and 4 s for association and 9 and 11's for ionic strength-induced dissociation at 20 degrees C. The interaction with the phospholipid component consists predominantly of a hydrophobic binding with the core of the membrane which may account for the membrane stabilizing effect of synapsin I. 4. Phosphorylation of synapsin I by either SV-associated or purified exogenous Ca2+/calmodulin-dependent protein kinase II (CaMPKII) inhibited the association rate and the binding to SV at steady state by acting on the ionic strength-sensitive component of the binding. When dephosphorylated synapsin I was previously bound to SV, exposure of SV to Ca2+/calmodulin in the presence of ATP triggered a prompt dissociation of synapsin I with a time constant similar to that of ionic strength-induced dissociation. 5. In conclusion, the reversible interactions between synapsin I and SV are highly regulated by site-specific phosphorylation and have kinetics of the same order of magnitude as the kinetics of SV recycling determined in mammalian neurons under comparable temperature conditions. These findings are consistent with the hypothesis that synapsin I associates with, and dissociates from, SV during the exo-endocytotic cycle. The on-vesicle phosphorylation of synapsin I by the SV associated CaMPKII, and the subsequent dissociation of the protein from the vesicle membrane, though not involved in mediating exocytosis of primed vesicles evoked by a single stimulus, may represent a prompt and efficient mechanism for the modulation of neurotransmitter release and presynaptic plasticity. PMID- 9401960 TI - Evidence of two mechanisms for the activation of the glucose transporter GLUT1 by anisomycin: p38(MAP kinase) activation and protein synthesis inhibition in mammalian cells. AB - 1. Inhibitors of protein synthesis stimulate sugar transport in mammalian cells through activation of plasma membrane GLUT1, the housekeeping isoform of the glucose transporter. However, it has been reported that some of these compounds, in addition to their effect on protein synthesis, also activate protein kinases. 2. In the present study we have explored the role of these two effects on GLUT1 activation. In 3T3-L1 adipocytes and Clone 9 cells, stimulation of sugar transport by puromycin, a translational inhibitor that does not activate kinases, was not detectable until 90 min after exposure. In contrast, stimulation by anisomycin, a potent Jun-NH2-terminal kinase (JNK) agonist, exhibited no lag phase. An intermediate response was observed to emetine and cycloheximide, weak activators of JNK. 3. The potency of anisomycin to stimulate transport acutely (30 min of exposure) was 5- to 10-fold greater than for its chronic stimulation of transport, measured after 4 h of exposure. The stimulation of transport by a low concentration of anisomycin (0.3 microM) was transient, peaked at 30-60 min and it was inhibited (IC50 < 1 microM) by SB203580, which indicates that its mediator is not JNK, but the homologous p38(MAP kinase) (p38(MAPK)). In contrast, the responses to 4 h exposure to 300 microM anisomycin or puromycin were refractory to SB203580. 4. Exposure to anisomycin resulted in rapid activation of p38(MAPK). Activation of both p38(MAPK) and GLUT1 by 0.3 microM anisomycin was cancelled by puromycin. 5. We conclude that the activation of GLUT1 in response to anisomycin includes two components: a delayed component involving translational inhibition and a fast, puromycin-inhibitable component that is secondary to activation of p38(MAPK). PMID- 9401961 TI - Leptin activates ATP-sensitive potassium channels in the rat insulin-secreting cell line, CRI-G1. AB - 1. Whole-cell current-clamp recordings demonstrate that leptin (0.3-10 nm) hyperpolarizes CRI-G1 insulin-secreting cells. This effect is slow on onset and is not reversed on washout of the leptin. 2. Voltage-clamp recordings indicate that leptin activates a potassium conductance in the presence of intracellular ATP (5 mm), but has not effect in its absence. Following activation of ATP sensitive K+ (KATP) current by diazoxide (0.2 mm), addition of leptin did not alter cell membrane potential or potassium current further. 3. The leptin-induced hyperpolarization and increased potassium conductance are completely inhibited by the application of the sulphonylureas tolbutamide (100 microM) and glibenclamide (0.5 microM). 4. Cell-attached and inside-out single-channel recordings indicate that leptin activates tolbutamide-sensitive KATP channels in CRI-G1 insulin secreting cells. PMID- 9401962 TI - Epoxyeicosatrienoic acids activate a high-conductance, Ca(2+)-dependent K + channel on pig coronary artery endothelial cells. AB - 1. Epoxyeicosatrienoic acids (EETs) have been described as endothelium-derived hyperpolarizing factors (EDHFs), based on their stimulatory effects on smooth muscle K+ channels. In order to reveal a putative autocrine effect of EETs on endothelial channels, we have studied the effects of the four EET regioisomers (5,6-EET, 8,9-EET, 11,12-EET and 14,15-EET) on the high-conductance, Ca(2+) dependent K+ (BKCa) channel recorded in inside-out patches of primary cultured pig coronary artery endothelial cells. Currents were recorded in the presence of either 500 nm or 1 microM free Ca2+ on the cytosolic side of the membrane. 2. In 81% of experiments, EETs at < 156 nM, applied on the cytosolic side of the membrane, transiently increased BKCa channel open state probability (PO) without affecting its unitary conductance, thus providing evidence for direct action of EETs, without involvement of a cytosolic transduction pathway. 3. The four EET regioisomers appeared to be equally active, multiplying the BKCa channel PO by a mean factor of 4.3 +/- 0.6 (n = 15), and involving an increase in the number and duration of openings. 4. The EET-induced increase in BKCa channel activity was more pronounced with low initial PO. When the BKCa channel was activated by 500 nM Ca2+, application of EETs increased the initial PO value of below 0.1 by a factor of 5. When the channel was activated by 1 microM Ca2+, application of EETs increased the initial PO value by a factor of 3. 5. Our results show that EETs potentiate endothelial BKCa channel activation by Ca2+. The autocrine action of EETs on endothelial cells, which occurs in the same concentration range as their action on muscle cells, should therefore fully participate in the vasoactive effects of EETs, and thus be taken into account when considering their putative EDHF function. PMID- 9401964 TI - A rapidly activating sustained K+ current modulates repolarization and excitation contraction coupling in adult mouse ventricle. AB - 1. The K+ currents which control repolarization in adult mouse ventricle, and the effects of changes in action potential duration on excitation-contraction coupling in this tissue, have been studied with electrophysiological methods using single cell preparations and by recording mechanical parameters from an in vitro working heart preparation. 2. Under conditions where Ca(2+)-dependent currents were eliminated by buffering intracellular Ca2+ with EGTA, depolarizing voltage steps elicited two rapidly activating outward K+ currents: (i) a transient outward current, and (ii) a slowly inactivating or 'sustained' delayed rectifier. 3. These two currents were separated pharmacologically by the K+ channel blocker 4-amino-pyridine (4-AP). 4-AP at concentrations between 3 and 200 microM resulted in (i) a marked increase in action potential duration and a large decrease in the sustained K+ current at plateau potentials, as well as (ii) a significant increase in left ventricular systolic pressure in the working heart preparation. 4. The current-voltage (I-V) relation, kinetics, and block by low concentrations of 4-AP strongly suggest that the rapid delayed rectifier in adult mouse ventricles is the same K+ current (Kv1.5) that has been characterized in detail in human and canine atria. 5. These results show that the 4-AP-sensitive rapid delayed rectifier is a very important repolarizing current in mouse ventricle. The enhanced contractility produced by 4-AP (50 microM) in the working heart preparation demonstrates that modulation of the action potential duration, by blocking a K+ current, is a very significant inotropic variable. PMID- 9401963 TI - Modulation of inwardly rectifying potassium channels in cultured bovine pulmonary artery endothelial cells. AB - 1. We have used the patch-clamp technique to study modulation of the inwardly rectifying K+ current (IK(IR)) in cultured bovine pulmonary artery endothelial cells (CPAE cells). In whole-cell mode, IK(IR) was defined as the Ba(2+) sensitive current. In single channel recordings, we observed a strongly inwardly rectifying and K(+)-selective channel with a conductance of 31 +/- 3 pS. 2. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis and functional data suggest that the endothelial IRK is most probably Kir2.1. 3. Intracellular ATP is required to prevent run-down of IRK in whole-cell mode. Single channel activity disappeared in inside-out patches exposed to ATP-free solution and in cell-attached patches on cells exposed to metabolic inhibition (KCN, 2 deoxyglucose). 4. The non-hydrolysable ATP analogues, ATP gamma S and adenylyl imidodiphosphate (AMP-PNP), did not prevent run-down. Run-down did not occur in the presence of okadaic acid, a phosphatase inhibitor, but was enhanced in the presence of protamine, an activator of phosphatase 2A (PP2A). 5. GTP gamma S and AlF4- inhibited IRK, also in the presence of ATP. GTP beta S antagonized the GTP gamma S effect. Pretreatment of the cells with PTX did not affect the GTP gamma S induced inhibition. Okadaic acid, however, slowed this inhibition. 6. Neither activation of protein kinase A (PKA) nor activation of protein kinase C (PKC) affected IRK. Additionally, neither cytochalasin B nor a high concentration of intracellular Ca2+ affected the time course of IRK run-down. 7. We conclude that run-down of IRK is probably due to dephosphorylation by PP2A. Activation of a PTX insensitive G protein inhibits this current by a mechanism that is neither mediated via the PKA and PKC pathways nor by intracellular Ca2+, but supposedly by a G protein-dependent activation of a phosphatase. PMID- 9401965 TI - Calcium-induced release of strontium ions from the sarcoplasmic reticulum of rat cardiac ventricular myocytes. AB - 1. The effects of strontium ions, Sr2+, on Ca(2+)-dependent feedback mechanisms during excitation-contraction coupling were examined in voltage-clamped rat ventricular myocytes in which intracellular [Ca2+] and [Sr2+] were monitored with the fluorescent indicator, indo-1. 2. Voltage clamp depolarizations and caffeine applications during superfusion in Ca(2+)-free, Sr(2+)-containing solutions were employed to exchange intracellular Ca2+ with Sr2+. Myocytes were loaded with Sr2+ by applying voltage clamp depolarizations during superfusion in Na(+)-free, Sr(2+)-containing solutions. 3. Caffeine applications produced large fluorescence transients in Sr(2+)-loaded cells. Thus, Sr2+ could be sequestered and released from the sarcoplasmic reticulum. 4. Ca2+ influx, but not Sr2+ influx, via sarcolemmal Ca2+ channels evoked ryanodine-sensitive fluorescence transients in Sr(2+)-loaded cells. These results demonstrated that Ca2+ influx-induced Sr2+ release (CISR) from the sarcoplasmic reticulum occurred in these experiments, even though Sr2+ influx-induced Sr2+ release was not observed. 5. The amplitude of the Ca2+ influx-induced fluorescence transient was 17 +/- 1% of the caffeine induced transient (n = 5 cells), an indication that fractional utilization of Sr2+ sequestered in the sarcoplasmic reticulum during CISR was low. 6. With increased Sr2+ loading, the amplitude of Ca2+ influx- and caffeine-induced fluorescence transients increased, but fractional utilization of sarcoplasmic reticulum divalent cation stores was independent of the degree of Sr2+ loading. These data suggest that Ca2+ influx directly activated the release of divalent cations from the sarcoplasmic reticulum, but mechanisms promoting positive feedback of Sr2+ release were minimal during CISR. 7. By comparison, in Ca(2+) loaded myocytes, Ca2+ influx-induced Ca2+ release (CICR) utilized a greater fraction of caffeine-releasable stores than CISR. Fractional utilization of Ca2+ stores during CICR increased with the degree of Ca2+ loading. 8. Taken together, these results suggest that Ca(2+)-dependent feedback mechanisms play a major role in determining the extent of sarcoplasmic reticulum Ca2+ release during cardiac excitation-contraction coupling under a wide range of conditions. PMID- 9401966 TI - Calcium homeostatic mechanisms operating in cultured postnatal rat hippocampal neurones following flash photolysis of nitrophenyl-EGTA. AB - 1. We examined Ca2+ homeostatic mechanisms in cultured postnatal rat hippocampal neurones by monitoring the recovery of background-subtracted fluo-3 fluorescence levels at 20-22 degrees C immediately following a rapid increase in Ca2+ levels induced by flash photolysis of the caged Ca2+ compound nitrophenyl-EGTA (NP EGTA). 2. A variety of methods or drugs were used in attempt to block specifically efflux of Ca2+ by the plasmalemmal Na(+)-Ca2+ exchanger or uptake of Ca2+ into mitochondria. 3. Many of the experimental manipulations produced a decrease in intracellular pH (pHi) measured in sister cultures using the pH sensitive dye 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein (BCECF). Accordingly, in each case, we determined the appropriate amount of the weak base trimethylamine (TMA) required to restore baseline pHi prior to flash photolysis. 4. Blockade of the plasmalemmal Na(+)-Ca2+ exchanger by replacement of external Na+ with either Li+ or N-methyl-D-glucamine (NMDG) markedly reduced pHi but did not affect the rate of recovery of fluo-3 fluorescence intensities once pHi was restored. 5. Inhibition of mitochondrial Ca2+ uptake, using the protonophore carbonyl cyanide m-chloro-phenylhydrazone (CCCP), resulted in a reduction in pHi, which could be restored by the addition of 2 mM TMA. Under these conditions the rate of recovery of Ca2+ levels was significantly slower than in the controls. Similar results were found using the respiratory chain inhibitor rotenone. 6. We conclude that, when the potential effects of changes in pHi are taken into account, mitochondria appear to sequester significant amounts of Ca2+ in the neuronal preparations used. PMID- 9401967 TI - A novel slow hyperpolarization-activated potassium current (IK(SHA)) from a mouse hippocampal cell line. AB - 1. A slow hyperpolarization-activated inwardly rectifying K+ current (IK(SHA)) with novel characteristics was identified from the mouse embryonic hippocampus x neuroblastoma cell line HN9.10e. 2. The non-inactivating current activated negative to a membrane potential of -80 mV with slow and complex activation kinetics (tau act approximately 1-7 s) and a characteristic delay of 1-10 s (-80 to -140 mV) that was linearly dependent on the membrane potential. 3. Tail currents and instantaneous open channel currents determined through fast voltage ramps reversed at the K+ equilibrium potential (EK) indicating that primarily K+, but not Na+, permeated the channels. 4. IK(SHA) was unaffected by altering the intracellular Ca2+ concentration between approximately 0 and 10 microM, but was susceptible to block by 5 mM extracellular Ca2+, Ba2+ (Ki = 0.42 mM), and Cs+ (Ki = 2.77 mM) 5. In cells stably transformed with M2 muscarinic receptors, IK(SHA) was rapidly, but reversibly, suppressed by application of micromolar concentrations of muscarine. 6. At the single channel level K(SHA) channel openings were observed with the characteristic delay upon membrane hyperpolarization. Analysis of unitary currents revealed an inwardly rectifying I V profile and a channel slope conductance of 7 pS. Channel activity persisted in the inside-out configuration for many minutes. 7. It is concluded that IK(SHA) in HN9.10e cells represents a novel K+ current, which is activated upon membrane hyperpolarization. It is functionally different from both classic inwardly rectifying IKir currents and other cationic hyperpolarization-activated IH currents that have been previously described in neuronal or glial cells. PMID- 9401968 TI - Functional nicotinic ACh receptors on interneurones in the rat hippocampus. AB - 1. Neuronal nicotinic ACh receptors (nAChRs) were studied in the rat hippocampal slice preparation using whole-cell patch-clamp recording techniques. 2. Responses to ACh (100 microM) were detected on inhibitory interneurones in the Ca1 field of the hippocampus proper and in the dentate gyrus, but not on principal excitatory neurones in either region. The different neuronal types were identified based on their morphology and location. 3. ACh excited interneurones in the hippocampus and dentate gyrus in current-clamp recordings. In voltage-clamp recordings, ACh activated inward currents were recorded from interneurones in the presence of blockers of synaptic transmission and the muscarinic ACh receptor antagonist atropine. The zero current potential for this response to ACh was near 0 mV. 4. The effect of ACh was mimicked by the nAChR-selective agonists nicotine (100 microM) and 1,1-dimethyl-4-phenyl-piperazinium iodide (DMPP, 100 microM). The response to ACh was reversibly antagonized by the neuronal nAChR antagonist mecamylamine (10 microM). The nAChR alpha 7 subunit-selective antagonists alpha bungarotoxin (100 nM) and methyllycaconitine (10 nM) also inhibited the response to ACh. 5. These observations demonstrate the presence of functional nAChRs on inhibitory interneurones in the rat hippocampus. Thus, a novel mechanism by which ACh can regulate neuronal activity in the hippocampus is revealed. PMID- 9401969 TI - Interchangeable discharge patterns of neurons in caudal nucleus tractus solitarii in rat slices: role of GABA and NMDA. AB - 1. We characterized in rat brain slices the discharge patterns of spontaneously active neurons in the caudal region of the nucleus tractus solitarii (cNTS) and the neuromodulatory role of GABA and glutamate, via GABAA and NMDA receptors. 2. Spontaneous action potentials recorded intracellularly from cNTS neurons manifested either a regular or an irregular discharge pattern, alongside characteristic waveforms of the action potentials. These discharge patterns were interchangeable, and were highly sensitive to fluctuations in membrane potentials. In addition, the repolarizing rate of the after-hyperpolarization (AHP) in cNTS neurons that exhibited a regular discharge pattern was significantly higher than that of neurons that displayed irregular discharges. 3. cNTS neurons that manifested a regular discharge pattern were converted to irregular discharges upon superfusion with GABA (200 microM). This was accompanied by a reduction in the repolarizing rate of the AHP of both spontaneous and evoked action potentials. Conversion of discharge patterns in the opposite direction was elicited by superfusion with NMDA (6.8 microM). 4. The irregular discharges of spontaneous or evoked cNTS neurons were converted to a regular discharge pattern by bicuculline (200 microM). Subsequent application of D(-)-2-amino-5-phosphonopentanoic acid (250 microM) essentially led the neuronal discharges to revert to an irregular pattern. 5. Our results support the presence of two interchangeable modes of electrophysiological manifestations from the same cNTS neuronal population. They also showed that GABA and glutamate, via GABAA and NMDA receptors, may provide a novel form of neuromodulation at the cNTS by switching the patterns of neuronal discharges. PMID- 9401970 TI - Different roles for GABAA and GABAB receptors in visual processing in the rat superior colliculus. AB - 1. The superficial grey layer of the superior colliculus (SGS) contains a high proportion of GABAergic inhibitory neurones. We have investigated the role of GABA receptors in synaptic transmission of aspects of visual activity in the SGS that may be driven by inhibitory mechanisms, such as surround inhibition and response habituation. 2. Multi-barrel glass iontophoretic pipettes were used to record single neuronal activity in the SGS of urethane-anaesthetized rats. Visual stimulation was provided by the display of moving bars and stationary spots of light on a monitor placed in the receptive field. 3. Both ejection of GABA and the GABAB agonist baclofen reduced responses to moving bars (interstimulus intervals > or = 8 s). The effects of GABA were reversed by the GABAA antagonist bicuculline, and the effects of baclofen were antagonized by the GABAB antagonist CGP 35,348. 4. Surround inhibition was estimated by plotting the response to flashed spots of increasing diameter. In controls, expanding the spot diameter beyond the excitatory receptive field caused a decrease in the response. This inhibitory surround was reversibly reduced by bicuculline, but CGP 35,348 had no effect. 5. Response habituation is the progressive reduction in the visual response during repetitive stimulus presentation. In controls, the visual response was reduced to 44 +/- 3% of its initial level when a stimulus (moving bar) was presented 5 times with an interstimulus interval of 0.5 s. During CGP 35,348 ejection, response habituation was reversibly reduced. Bicuculline had no effect on response habituation. 6. The effects of bicuculline on surround inhibition in the superior colliculus are consistent with similar studies in the lateral geniculate nucleus which indicate that GABAA receptors mediate this effect. The function of GABAB receptors in the visual system is less well researched. The reduction of response habituation with CGP 35,348 demonstrates that, at least in the SGS, GABAB receptors have an important role in visual transmission which is distinct from that of GABAA receptors. PMID- 9401971 TI - Novel glutamate- and GABA-independent synaptic depolarization in granule cells of guinea-pig hippocampus. AB - 1. Dual intracellular recordings of granule cells, hilar interneurons and CA3 pyramidal cells were performed in transverse slices of guinea-pig hippocampus. At resting membrane potential, in the presence of 4-aminopyridine, ionotropic glutamate receptor antagonists and the GABAA receptor antagonist bicuculline, granule cells showed spontaneous, large amplitude depolarizations correlated with synchronous bursting activity of interneurons. 2. Under these conditions, pyramidal cells exhibited large amplitude monophasic GABAB inhibitory postsynaptic potentials (IPSPs) synchronous with the GABAergic interneuron burst discharges. The granule cells also received a GABAB input, which was evident only when the neurons were depolarized by DC injection. The GABAB receptor antagonist CGP 55,845A (CGP) blocked the GABAB IPSPs in both pyramidal cells and granule cells; however, the depolarizing potential in granule cells was unaffected by the drug. 3. The granule cells depolarization in the presence of CGP was monophasic and exhibited linear voltage dependence with a reversal potential around -40 mV, suggesting that it was generated by a synaptic input activating a mixed cationic current. 4. The granule cell depolarization was abolished following the addition of tetrodotoxin to the bath. In addition, perfusing the slice with a low Ca(2+) containing solution (0.5 mM Ca(2+)-10 mM Mg2+) also abolished the granule cell depolarization, confirming the synaptic origin of the event. 5. (S)-Methyl-4 carboxyphenylglycine, L-(+)-2-amino-3-phosphonopropionic acid, propranolol and atropine did not affect the granule cell depolarization, indicating that metabotropic glutamate receptors, beta-adrenergic receptors and muscarinic cholinergic receptors were not involved in generating the granule cell depolarizing synaptic response. 6. These findings indicate that, in the absence of both glutamatergic and GABAergic inputs, synchronous interneuronal activity can produce a depolarizing synaptic response in granule cells. The neurochemical responsible for the depolarization is currently under investigation. PMID- 9401972 TI - Characterization of the inward current induced by metabotropic glutamate receptor stimulation in rat ventromedial hypothalamic neurones. AB - 1. Whole-cell patch clamp recordings were made from rat ventromedial hypothalamic neurones in slices of brain tissue in vitro. Bath application of 50 microM (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid (1S,3R-ACPD) depolarized all neurones tested by activation of an inward current of approximately 55 pA at -60 mV. 2. The inward current elicited by 1S,3R-ACPD was unaffected by K+ channel blockade with external Cs+, Ba2+ or TEA. However, the current was significantly reduced by replacement of the external NaCl with either Tris-HCl or LiCl. 3. The 1S,3R-ACPD-induced current was reduced by the heavy metal ions Ni2+ or La3+ and also by the Na(+)-Ca2+ exchange current inhibitor 3',4'-dichlorobenzamil. 4. The effects of 1S,3R-ACPD were mimicked by the group I metabotropic agonist 3,5 dihydroxyphenylglycine (DHPG) but not by the group III selective agonist, L-2 amino-4-phosphonobutanoate (L-AP4). Furthermore, the effects of 1S,3R-ACPD were inhibited by the metabotropic antagonists alpha-methyl-4-carboxyphenylglycine (MCPG) and 1-aminoindan-1,5-dicarboxylic acid (AIDA) but not by the presynaptic metabotropic receptor antagonists alpha-methyl-4-phosphonophenylglycine (MPPG) or alpha-methyl-4-tetrazolylphenylglycine (MTPG). 5. Photorelease of caged GDP beta S inside neurones irreversibly blocked the 1S,3R-ACPD-induced current whilst photolysis of caged GTP gamma S inside neurones irreversibly potentiated this current. 6. The PLC inhibitor U-73,122 significantly reduced the size of the inward current induced by 1S,3R-ACPD. This effect was not mimicked by the inactive analogue U-73,343. 7. Flash photolysis of the caged calcium chelator diazo-2 inside neurones diminished the response to 1S,3R-ACPD. 8. It is concluded that group I metabotropic glutamate receptors depolarize neurones in the VMH by activation of a Na(+)-Ca2+ exchange current through a G-protein coupled increase in intracellular Ca2+. PMID- 9401973 TI - Age-related functional changes of the glutamate receptor channels in rat Meynert neurones. AB - 1. The developmental changes of glutamate receptors (GluRs) in acutely dissociated rat Meynert neurones were investigated using the conventional whole cell and nystatin perforated patch recording modes under voltage-clamp conditions. 2. The neurones became less responsive to N-methyl-D-aspartic acid (NMDA) with age, most dramatically between 1 day and 2 weeks, while the responses to kainic acid (KA) and L-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) gradually increased. The metabotropic GluR response appeared a few days after birth, but thereafter no further change was observed. 3. The decrease in the NMDA response during postnatal development was due to an abrupt reduction in the number of receptors without affecting the affinity, voltage-dependent Mg2+ blockade or high Ca2+ permeability (PCa/PCs approximately 7.0). 4. PCa/PCs in the presence of KA decreased from 2.8 in the 1-day-old (1D) rat neurones to 1.1 and 0.44 in the 2-week-old (2W) and 6-month-old (6M) rat neurones, respectively. The concentration-response relationship for KA shifted to the left with age. The KA response was not affected by NS-102, a KA-selective antagonist, thus indicating that the increased affinity of the receptor for the ligand resulted from the change in the AMPA receptor channel subunits. 5. The AMPA response in the presence of 10(-4) M cyclothiazide showed a change in the inward rectifying current-voltage relationship with age. The KA response was strongly cross desensitized by the addition of AMPA and was also blocked by 6-cyano-7 nitroquinoxaline-2,3-dione (CNQX), whereas a rapid desensitization of the AMPA response was removed in a concentration-dependent manner by cyclothiazide. These results indicate that the non-NMDA receptor channels are assembled from the subunits of the AMPA receptor family without the GluR-2 subunit, thus resulting in a high Ca2+ permeability. 6. The L-glutamate (Glu)-induced responses were more sensitive to DL-2-amino-5-phosphonopentanoic acid (APV) in the 1D rat neurones than in the adult rat neurones. 7. Both NMDA and KA raised the intracellular Ca2+ concentration ([Ca2+]i) in all neurones of 1D, 2W and 6M rats, though the charybdotoxin-sensitive Ca(2+)-activated K+ current (IK(Ca)) did not appear in the 1D rat neurones. An age-related prolongation of both IK(Ca) decay and [Ca2+]i clearance was also seen after the removal of KA. 8. It was thus concluded that the age-related changes of ionotropic receptors appear to play a key role in the activities of immature and mature rat Meynert cholinergic neurones. The KA induced IK(Ca), which developed with ageing, may thus function as one of the negative feedback systems, and thereby prevent excess cell excitation and neural damage, especially in adult rats. PMID- 9401974 TI - Glycine receptors in cultured chick sympathetic neurons are excitatory and trigger neurotransmitter release. AB - 1. Total RNA isolated from embryonic chick paravertebral sympathetic ganglia was used in a reverse transcription-polymerase chain reaction (RT-PCR) assay with a pair of degenerate oligonucleotide primers deduced from conserved regions of mammalian glycine receptor alpha-subunits. Three classes of cDNA were identified which encode portions of the chicken homologues of the mammalian glycine receptor alpha 1, alpha 2 and alpha 3 subunits. 2. The presence of functional glycine receptors was investigated in the whole-cell configuration of the patch-clamp technique in neurons dissociated from the ganglia and kept in culture for 7-8 days. In cells voltage clamped to -70 mV, glycine consistently induced inward currents in a concentration-dependent manner and elicited half-maximal peak current amplitudes at 43 microM. 3. The steady-state current-voltage relation for glycine-induced currents was linear between +80 and -60 mV, but showed outward rectification at more hyperpolarized potentials. Reversal potentials of these currents shifted with changes in intracellular chloride concentrations and matched the calculated Nernst potentials for chloride. 4. beta-Alanine and taurine were significantly less potent than glycine in triggering inward currents, with half-maximal responses at 79 and 86 microM, respectively. At maximally active concentrations, beta-alanine-evoked currents were identical in amplitude to those induced by glycine. Taurine-evoked currents, in contrast, never reached the same amplitude as glycine-induced currents. 5. The classical glycine receptor antagonist strychnine reversibly reduced glycine-induced currents, with half-maximal inhibition occurring at 62 nM. Two more recently characterized glycine receptor antagonists, isonipecotic acid (half-maximal inhibition at 2 mM) and 7-trifluoromethyl-4-hydroxyquinoline-3-carboxylic acid (half-maximal inhibition at 67 microM), also blocked glycine-evoked currents in a reversible manner. The chloride channel blocker picrotoxin reduced glycine-evoked currents, with half-maximal effects at 348 microM. Inhibition by the glycine receptor channel blocker cyanotriphenylborate was half-maximal at 4 microM. 6. Apart from evoking inward currents, glycine occasionally triggered short (< 100 ms) spike-like currents which were abolished by hexamethonium and thus reflected synaptic release of endogenous acetylcholine. In addition, glycine caused Ca(2+) dependent and tetrodotoxin-sensitive tritium overflow from neurons previously labelled with [3H]noradrenaline. This stimulatory action of glycine was reduced in the presence of strychnine and after treatment with the chloride uptake inhibitor furosemide (frusemide). 7. In 65% of neurons loaded with the Ca2+ indicator fura-2 acetoxymethyl ester, glycine increased the ratio of the fluorescence signal obtained with excitation wavelengths of 340 and 380 nm, respectively, which indicates a rise in intracellular Ca2+ concentration. 8. The results show that sympathetic neurons contain transcripts for different glycine receptor alpha-subunits and carry functional heteromeric glycine receptors which depolarize the majority of neurons to trigger transmitter release. PMID- 9401975 TI - Adenosine formation in contracting primary rat skeletal muscle cells and endothelial cells in culture. AB - 1. The present study examined the capacity for adenosine formation, uptake and metabolism in contracting primary rat muscle cells and in microvascular endothelial cells in culture. 2. Strong and moderate electrical simulation of skeletal muscle cells led to a significantly greater increase in the extracellular adenosine concentration (421 +/- 91 and 235 +/- 30 nmol (g protein) 1, respectively; P < 0.05) compared with non-stimulated muscle cells (161 +/- 20 nmol (g protein)-1). The ATP concentration was lower (18%; P < 0.05) in the intensely contracted, but not in the moderately contracted muscle cells. 3. Addition of microvascular endothelial cells to the cultured skeletal muscle cells enhanced the contraction-induced accumulation of extracellular adenosine (P < 0.05), whereas endothelial cells in culture alone did not cause extracellular accumulation of adenosine. 4. Skeletal muscle cells were found to have ecto-forms of several enzymes involved in nucleotide metabolism, including ATPases capable of converting extracellular ATP to ADP and AMP. 5. Adenosine added to the cell medium was taken up by muscle cells and incorporated into the adenine nucleotide pool so that after 30 min of incubation, over 95% of the adenosine label was present in ATP, ADP and AMP. A similar extent of incorporation of adenosine into the nucleotide pool was evident in the endothelial cells. 6. The present data suggest that contracting muscle cells induce an elevation in the extracellular adenosine concentration. Addition of endothelial cells to muscle cells enhances the contraction-induced formation of adenosine. Adenosine taken up by muscle and endothelial cells from the extracellular space is not likely to be used for storage in intracellular pools, but may serve to regulate muscle extracellular adenosine levels. PMID- 9401976 TI - Modulation of stimulus-secretion coupling in single rat gonadotrophs. AB - 1. Exocytosis and intracellular [Ca2+] were determined simultaneously in single anterior pituitary gonadotrophs from ovariectomized female rats. Dispersed cells were cultured for 2-4 days with or without 0.2 nM oestradiol-17 beta (E2) before use. Cells were stimulated with either gonadotrophin releasing hormone (GnRH) or by membrane depolarization. Exocytosis was determined from the change in membrane capacitance (Cm) using the perforated-patch whole-cell recording technique. Intracellular [Ca2+] was measured using fura-2 fluorescence. 2. The exocytotic response to 1 nM GnRH was characterized by a wide spectrum of responses, ranging from exocytotic bursts to relatively slow, graded increases that were dependent on the evoked intracellular Ca2+ pattern. A kinetic model is presented that incorporates the observed steep dependence of exocytosis on measured intracellular [Ca2+]; simulated exocytosis reasonably approximated observed exocytotic responses, both kinetically and quantitatively. The model also suggests that the modulatory effects of E2 are brought about either by a change in the Ca2+ sensitivity of exocytosis or by a preferential clustering of docked secretory granules close to sites of Ca2+ release. The results suggest that in gonadotrophs an oscillatory Ca2+ signal is sensed by the exocytotic apparatus in a modified form of digital encoding. 3. Exocytosis in E2-treated cells was 3-fold greater than in non-treated cells for GnRH-evoked secretion, and 38% greater for depolarization; however, there was no effect of E2 on the intracellular Ca2+ response to either stimulus. The results show that maximum expression of the effect of E2 on exocytosis requires activation of GnRH-dependent pathways. PMID- 9401977 TI - Extracellular glucose turnover in the striatum of unanaesthetized rats measured by quantitative microdialysis. AB - 1. Steady-state and time-resolved quantitative microdialysis was used to measure dialysate concentration, extracellular concentration and the in vivo recovery of glucose in rat striatum. 2. The extracellular concentration of glucose, determined by the zero net flux method of Lonnroth, was 350 +/- 20 microM and the in vivo recovery was 39 +/- 2%. 3. Veratridine caused a steep decrease in dialysate glucose after an initial delay of 7.5 min. When steady-state glucose levels had been reached in the presence of veratridine the extracellular concentration was reduced to zero, but there was no significant change in in vivo recovery. 4. Measurement of the dynamic changes during the administration of veratridine showed an immediate decrease in extracellular glucose concentration and a steep rise in in vivo recovery, which accounted for the delay in the delay in the decrease in dialysate glucose. When extracellular concentration reached zero, in vivo recovery returned to control levels. PMID- 9401978 TI - Phasic activity in the human erector spinae during repetitive hand movements. AB - 1. Phasic activity in the human back muscle erector spinae (ES) was studied during repetitive hand movements. The hand movements were elicited voluntarily by the subject or induced passively by the experimenter through a servomotor or through cyclical electrical stimulation of muscles acting about the wrist. The aim of the study was to determine whether the rhythmical activation of ES was of supraspinal, intersegmental or segmental origin. 2. When voluntary rhythmical hand movements were performed as fast as possible, cyclical ES EMG bursts occurred at the same frequency. This frequency was significantly higher than that reached when the task was to contract the back muscles as rapidly as possible. This suggests that the ES activity during the fast hand movements was not generated by direct commands descending to the ES muscles from the motor area of the cerebral cortex responsible for voluntary back muscle activation. 3. During imposed rhythmical hand movements, ES EMG bursts remained entrained to the hand movements, even when movement frequencies far exceeded those attainable voluntarily either for the hand or the back. This showed that ES EMG responses could be evoked by the hand movements even when these were not generated by descending neural commands. Two alternative mechanisms of ES activation were considered: (a) propriospinal transmission of afferent input entering the spinal cord from the upper extremity; (b) afferent input from ES and other trunk muscles, responding to local oscillations transmitted mechanically from the hand to the lower back. 4. Activation of ES via proprioceptive signals from the forearm was unlikely since (a) simultaneous electrical stimulation of wrist extensor and wrist flexor muscles did not result in repetitive ES EMG bursting; (b) cyclical vibration of the wrist extensors did not evoke ES EMG bursting; (c) when the forearm was constrained and the hand was moved passively, the lower trunk accelerations and cyclical ES EMG both occurred at a harmonic of the hand movement frequency. 5. We conclude that the repetitive ES EMG bursting during hand movements was probably due to a local segmental reflex rather than to descending commands. Remote mechanical oscillations of the trunk caused by hand movements evidently elicited proprioceptive reflexes in ES that presumably contributed to trunk stabilization. PMID- 9401979 TI - Firing pattern of type-identified wrist extensor motor units during wrist extension and hand clenching in humans. AB - 1. Single motor unit activity was investigated in the extensor carpi radialis muscles during voluntary isometric contraction involving either the coactivation of the wrist agonist extensor muscles (wrist extension) or the coactivation of the wrist and finger antagonist extensor and flexor muscles (hand clenching). 2. The motor units were found to be activated at a similar level of motoneurone pool drive during both wrist extension and hand clenching, as indicated by the fact that the EMG activity at which they were recruited was practically the same in both cases (mean +/- S.D.: 20 +/- 26 and 21 +/- 25 mV, respectively). In addition, the net excitatory drive exerted on the motoneurones, as assessed from the mean interspike intervals, did not differ significantly between the two tasks (mean +/- S.D.: 104.57 +/- 17.24 and 103.01 +/- 16.26 ms, for wrist extension and hand clenching, respectively). 3. However, the discharge variability, in terms of the coefficient of variation of the interspike intervals, was slightly but significantly greater during hand clenching than during wrist extension (0.213 +/ 0.049 and 0.198 +/- 0.045, respectively). This increase involved all types of motor units, regardless of their contractile force. 4. We suggest that the greater motoneurone discharge variability observed during hand clenching may be attributable to an increase in the synaptic noise. This increase might be due to the activation of numerous afferent pathways mediating reciprocal interactions between antagonist motoneurone pools, as well as to the activation of hand cutaneous receptors that play a major role in the regulation of handling and gripping motor activities. PMID- 9401981 TI - Cross-facial vein grafting in complicated flap reconstructions of the face and mandible. AB - Microvascular flap reconstructions of facial and mandibular defects are achieving increasingly widespread application and, as a result, more patients are considered for such procedures after large tumors, congenital abnormalities, previous surgery (including failed and successful microsurgical procedures) and radiation have compromised potential recipient vessels on the side of the defect. In three such cases, contralateral facial vessels with vein grafts have been used to provide flap-recipient vessels. Each flap was successful. Cross-facial vein grafting is a relatively simple solution for utilizing undamaged vessels as recipient pedicles in complicated flap reconstructions of the face and mandible. PMID- 9401980 TI - Reinterpretation of endothelial cell gaps induced by vasoactive mediators in guinea-pig, mouse and rat: many are transcellular pores. AB - 1. In response to vascular permeabilizing agents, particulates circulating in the blood extravasate from venules through endothelial cell openings. These openings have been thought to be intercellular gaps though recently this view has been challenged. 2. To define the precise location of endothelial cell gaps, serial section electron microscopy and three-dimensional reconstructions were performed in skin and cremaster muscle of guinea-pigs, mice and rats injected locally with agents that enhance microvascular permeability: vascular permeability factor, histamine or serotonin. Ferritin and colloidal carbon were injected intravenously as soluble and particulate macromolecular tracers, respectively. 3. Both tracers extravasated from venules in response to all three permeability enhancing agents. The soluble plasma protein ferritin extravasated primarily by way of vesiculo vacuolar organelles (VVOs), interconnected clusters of vesicles and vacuoles that traverse venular endothelium. In contrast, exogenous particulates (colloidal carbon) and endogenous particulates (erythrocytes, platelets) extravasated from plasma through transendothelial openings. 4. Serial electron microscopic sections and three-dimensional reconstructions demonstrated that eighty-nine of ninety-two openings were transendothelial pores, not intercellular gaps. Pore frequency increased 3- to 33-fold when carbon was used as tracer. 5. The results demonstrate that soluble and particulate tracers extravasate from venules by apparently different transcellular pathways in response to vasoactive mediators. However, some pores may derive from rearrangements of VVOs. PMID- 9401982 TI - Plasticity and function--the fate of a free, neurovascular muscle graft ten years post-reconstruction. AB - A 16-year-old female sustained a subtotal amputation of the left thigh. Debridement resulted in a bone and soft-tissue defect of 20 cm in length. The whole quadriceps muscle was lost, and the knee joint was open. The femur was stabilized by transfer of corticocancellous bone grafts. A latissimus dorsi muscle was harvested and transferred to reconstruct the lost quadriceps muscle. The thoracodorsal nerve was coaptated to the motor branch of the femoral nerve. The years after trauma, the muscle provides excellent motor function. EMG evaluation reveals no sign of denervation; macro-electromyography reveals only a moderate enlargement of motor units. There is recruitment of all motor units. Maximum voluntary torque of the transplanted muscle has decreased, compared to the contralateral rectus femoris. Histologic evaluation demonstrates a normal skeletal muscle with typical fiber distribution. These results indicate complete adaptability of the muscle at an atypical site, with a high degree of functional and structural plasticity of the skeletal muscle. The decreased voluntary torque of the transferred latissimus dorsi depends on the lower, total-fiber, cross sectional area--the result of a parallel fiber structure. PMID- 9401983 TI - Successful microsurgical tissue transfer in a patient with postsplenectomy thrombocytosis treated with platelet-phoresis. AB - With the use of platelet-phoresis, two microsurgical free-tissue transfers were successfully undertaken in a patient with postsplenectomy thrombocytosis that had initially caused flap failure. Rapid reduction in the platelet count allowed free tissue transfer in this patient who required early wound coverage. PMID- 9401984 TI - Vascularized fibula graft for spinal fusion in severe cervical kyphosis due to neurofibromatosis. AB - A successful anterior spinal fusion using vascularized fibula graft in the case of a 17-year-old male with severe cervical kyphosis due to neurofibromatosis is reported. Anterior spinal fusion at C2-7 using the fibula graft was performed with spinous process wiring. The kyphosis was corrected from 85 to 38 degrees. Bony fusion was obtained in 5 months without loss of correction. PMID- 9401985 TI - Macrovascular surgery and the microsurgeon. AB - The improvement of success rates in microsurgery can be attributed as much to better technical skills, as to the more frequent selection of donor or recipient sites with consistent, larger-caliber vessels. Often, these vessels may be larger than major limb source vessels, and anastomoses using loupes can then be successful, even without requiring an operating microscope Thus, distinguishing our capabilities from the domain of the general vascular surgeon, who traditionally deals only with the ravages of disease or trauma to such large vessels, has become blurred. For some free-tissue transfers, and especially limb replantations, perhaps it would be appropriate for the microsurgeon sometimes to enter the realm of the macrovascular surgeon for enhancement of the overall outcome. A review of our 202 free flaps and pediatric limb revascularizations has validated this opinion, as significant portions in 19 of these cases required unequivocal macrovascular surgery. These included vein-graft bypasses (9) of major segmental arterial defects of limbs (that incidentally improved collateral circulation, although intended primarily to simplify arterial inflow to a free flap simultaneously needed to cover a concomitant soft-tissue defect). Similarly, arterial grafts as part of a "flow-through" free flap (3) were used for immediate coverage and concurrent limb revascularization. Finally, two toddlers who sustained disruption of named leg vessels had microsurgical repair after referral from the vascular service; they believed we were better able to deal with such diminutive vasculature. These observations are not intended as evidence that vascular surgery may be better performed by the microsurgeon; rather, that the best results of microsurgery often will incorporate technical aspects usually considered as macrovascular surgery. PMID- 9401986 TI - Magnetic resonance imaging assessment of a microvascular anastomotic device for ferromagnetism. AB - Microvascular free-tissue transfers have assumed particular importance as reconstructive techniques of choice in centers where ablative surgery for primary and recurrent malignant disease is a focus. In the context of malignant disease, issues of surveillance for recurrence are paramount. As clinical experience with the diagnostic imaging characteristics of flap reconstructions has been acquired, magnetic resonance imaging (MRI) has assumed a prominent role in the evaluation for recurrent malignant disease. This has provided an important supportive role for contemporary concepts of immediate reconstruction. The Precise-TM Microvascular Anastomotic Device (MACD) is based on the friction-fit union of implant rings composed of high-density polyethylene and surgical stainless steel. Many characteristics of the device have been described in histologic and laboratory studies. As yet uncharacterized is the effect of clinical MRI electromagnetic fields on the device, which is composed, in part, of type 316 stainless steel. The MACD is in wide use in centers where microsurgeons are experienced with the system and it is designed to facilitate the performance and reliability of microvascular anastomoses. The implications for MRI as a safe imaging modality for the acute perioperative evaluation of patients reconstructed with microvascular free flaps anastomosed with the MACD are obvious. MACD implants of varying sizes were evaluated for displacement in each of three orthogonal planes within a 1.5 Tesla magnetic field. No change in displacement was observed for any of the devices. Magnetic resonance imaging may thus be considered a safe imaging modality for the acute perioperative diagnostic imaging of free-tissue transfers that have been anastomosed with the MACD. PMID- 9401987 TI - Primary flap closure in complex limb injuries. AB - Free-tissue transfers enable surgeons to reconstruct or salvage limbs injured or amputated in high-energy traumas which result in extensive damage to soft tissue, bone, tendons, vessels and nerves. Primary free-tissue transfer is performed following debridement, bone fixation, and repair of injured structures within 24 hr after injury. Between 1987 and 1996, 57 patients who had complex extremity traumas were treated with primary free-tissue transfer, or free flaps. Long-term follow-up ranged from 4 months to 9 years (median: 4.5 years). No flap failure or serious wound-healing complication occurred using the protocol. Radical debridement and primary free-flap coverage in extensive extremity injuries can salvage limbs, provide improved functional and aesthetic results, and psychologically benefit patients through lowered morbidity. Other benefits include reduced incidence of free-flap failure, postoperative infection, secondary operative procedures, and invalidity, as well as shorter hospital stays, and lowered medical expenses. PMID- 9401988 TI - Adjunctive measures in microvascular surgery. AB - Consistently good results following free-tissue transfers require not only basic skills in small vessel anastomosis, but also the implementation of numerous adjunctive measures. The pioneers in microsurgery have shared many of their hard learned lessons and clinical nuggets through texts, papers, and seminars. This article presents an assemblage of adjunctive measures of proven utility. PMID- 9401989 TI - Is axonal sprouting able to traverse the conjunctival layers of the peripheral nerve? PMID- 9401990 TI - Transrectal ultrasonographically assisted radical retropubic prostatectomy. AB - Radical prostatectomy is associated with difficulty in determining the division site of the urethra adjacent to the apical region of the prostate. We present the results of a feasibility clinical trial in three patients to determine whether intraoperative transrectal ultrasonography may assist during radical retropubic prostatectomy. Using this technique the apex is readily identified and a detailed view of the urethral stump could be obtained. In one case residual apical tissue was identified and excised. The creation of the vesicourethral anastomosis was documented and its water tightness was demonstrated. We conclude that transrectal ultrasonography can be performed during radical retropubic prostatectomy and may be helpful in selected cases. PMID- 9401991 TI - Sonographic imaging of the trachea. AB - We evaluated the feasibility of ultrasonography for imaging of the trachea and its effectiveness in the diagnosis and follow-up of patients with tracheal stenosis due to various diseases. Twenty normal volunteers and six adult patients with tracheal stenosis were included in the study group. Subjects were examined with ultrasonography in a supine position with the neck hyperextended or in a sitting position. At the level of the thyroid isthmus, the anterior tracheal wall thicknesses imaged by ultrasonography were 1.54 +/- 0.22 mm (mean +/- SD) and 1.22 +/- 0.18 mm for normal male and female volunteers, respectively. Ultrasonography could reveal the intrinsic tracheal wall lesions and extrinsic lesions compressing the trachea in patients with tracheal stenosis. These ultrasonographic images correlated with CT images. In conclusion, ultrasonography may be useful in imaging of the trachea. PMID- 9401992 TI - Sonographic, magnetic resonance imaging, and mammographic assessments of preoperative size of breast cancer. AB - High resolution sonographic (39 cases), magnetic resonance imaging (32 cases), and mammographic (35 cases) measurements of preoperative size of breast cancer were correlated with the pathologic size in 39 patients with breast carcinoma to determine the most accurate imaging technique for breast cancer size. There were nine T1, 21 T2, four T3, and four T4 tumors. Sonographic and magnetic resonance imaging measurements of tumor size demonstrated correlation coefficients of 0.92 and 0.93, respectively, both of which were superior to that of mammography (0.84). Sonographic tumor size evaluation thus is shown to be equivalent to magnetic resonance imaging in this study. Three of nine (33%), four of seven (57%), and four of eight (50%) T1 tumors would have been overstaged by ultrasonography, magnetic resonance imaging, and mammography, respectively. Three of 21 (14.3%), one of 16 (6.3%), and two of 18 (11.1%) T2 tumors would have been understaged by ultrasonography, magnetic resonance imaging, and mammography, respectively. We therefore found ultrasonography to be of value in the diagnosis and staging of breast cancer. PMID- 9401993 TI - Birthweight prediction by three-dimensional ultrasonographic volumes of the fetal thigh and abdomen. AB - Our validation study examined a three-dimensional ultrasonographic phantom that contained irregularly shaped volume targets ranging from 0.5 to 76.1 milliliters. Four different examiners made blinded measurements from volume datasets that were acquired by 4 and 7 MHz transducers. Birthweight predictions using abdominal and thigh volumes from 18 term fetuses also were compared with two-dimensional ultrasonographic methods. In vitro volume measurements were accurate, precise, and repeatable despite a small systematic overestimation with increasing object size. Mean systematic error and precision for birthweight predictions by three dimensional ultrasonography (-0.03% +/- 6.1%) were not significantly different from those by two-dimensional ultrasonography (-0.60% +/- 8.8%). Conventional prediction methods yielded three birthweights with greater than 15% error. By comparison, except for one infant whose birthweight indicated an 11% error, all predictions based on fetal volume parameters were within 10% of true values. Accurate birthweight predictions by fetal volume parameters appear to be technically feasible at term gestation although their practical clinical application requires further investigation. Birthweight predictions in this manner may allow remote consultants to evaluate the fetus over wide-area computer networks despite the physical absence of the patient. PMID- 9401994 TI - Echogenicity of hepatic versus portal vein walls revisited with histologic correlation. AB - The portal vein wall typically is hyperechoic over a wide range of beam-vessel angles, whereas the hepatic vein wall is hyperechoic only when the incident beam and the vessel are perpendicular. This has been attributed to marked discrepancies in mural thickness, collagen content, or perivascular fat between portal and hepatic veins. We evaluated histologically the walls of portal and hepatic veins using three cadaveric livers. For vessels with luminal diameter above 2 to 3 mm, hepatic vein and portal vein wall thicknesses were similar such that portal vein walls were not more than 50% thicker than those of hepatic veins of comparable size. Hepatic vein walls were mostly composed of parallel, tightly packed collagen fibers. In contrast, portal vein walls were composed of loosely arrayed, nonparallel connective tissue fibers which were separated by multiple intervening spaces and only a minority of which were collagenous. Perivascular fat was not identified adjacent to intrahepatic vessels beyond the liver hilus. The marked differences in echogenicity between portal vein and hepatic vein walls typically observed at ultrasonography thus cannot be attributed to differences in mural thickness, collagen content, or perivascular fat between these vessels. Rather, the distinct composition of the hepatic vein wall renders it a specular reflector, which is hyperechoic only when the angle between the ultrasound beam and the vessel wall is close to 90 degrees, whereas the composition of the portal vein wall enables it to appear hyperechoic at a wide range of beam-vessel angles. PMID- 9401995 TI - Hemodynamic changes in the early phase of artificially created arteriovenous fistula: color Doppler ultrasonographic findings. AB - The objective of this study was to evaluate hemodynamic variables in arteriovenous fistulas by color Doppler ultrasonography. This study involved 28 patients with chronic renal failure who were sent to surgery clinic for creation of an arteriovenous fistula of the Brescia-Cimino type. Patients were evaluated preoperatively and on the first and seventh days postoperatively by a color Doppler ultrasound machine with a 7.5 MHz linear probe. The distal radial artery was examined preoperatively and the fistula itself postoperatively. Changes in the fistula size and the velocity, volume, and resistive index of the distal radial artery were recorded. Postoperatively the radial artery diameter, systolic flow rates, and volume flow had increased significantly, especially on the first day, in comparison to preoperative values. Resistive index values has decreased significantly at both the first and the seventh days postoperatively. Color Doppler ultrasonography is a very effective method in the evaluation of hemodynamics of arteriovenous fistulas in hemodialysis patients. It will allow an understanding of the pathology in nonfunctioning fistulas or of the cause of complications that develop secondarily. PMID- 9401996 TI - Blood flow in functional cysts and benign ovarian neoplasms in premenopausal women. AB - To assess the value of transvaginal color Doppler sonography in the differentiation of functional cysts from benign ovarian neoplasms in premenopausal women, 100 premenopausal women with the diagnosis of adnexal mass were enrolled in a prospective study. All patients underwent transvaginal color Doppler sonography during the follicular phase. We evaluated 107 masses. Tumor volume and morphology were assessed, as were tumor blood flow location, the number of vessels, the resistive and pulsatility indices, and the peak systolic velocity. Patients were followed up after 8 to 10 weeks by transvaginal sonography. Functional cysts were considered when spontaneous resolution occurred. Surgery was performed if a tumor enlarged or persisted after two scans. Thirty-nine (36.5%) cysts regressed spontaneously and 68 (63.5%) were removed surgically. Seven of the latter were follicular or luteal cysts and were considered to be functional cysts. No carcinoma was found. Arterial blood flow was detected in 28 (60.8%) functional cysts and in 42 (68.8%) benign neoplasms (P = 0.3446). The vessels were located peripherally in 27 (94.6%) functional cysts and in 37 (88.1%) benign neoplasms (P = 0.2226). No differences were found between functional cysts and benign neoplasms in mean resistive index (0.65, 95% confidence interval: 0.59 to 0.71 versus 0.64, 95% confidence interval: 0.60 to 0.69), mean pulsatility index (1.47, 95% confidence interval: 1.17 to 1.84 versus 1.57, 95% confidence interval: 1.26 to 1.86), number of vessels (1.1, 95% confidence interval: 0.7 to 1.3 versus 1.4, 95% confidence interval: 1.1 to 1.8), and peak systolic velocity (28.6 cm/s, 95% confidence interval: 24.7 to 34.2 versus 24.9 cm/s, 95% confidence interval: 21.6 to 28.3). We concluded that transvaginal color Doppler sonography is not useful to discriminate between functional ovarian cysts and benign ovarian neoplasms in premenopausal women. PMID- 9401997 TI - Color Doppler ultrasonography in the diagnosis of portal vein invasion in patients with pancreatic cancer. AB - We retrospectively evaluated the diagnostic usefulness of color Doppler ultrasonography for detecting portal vein invasion in 21 patients with pancreatic cancer who underwent surgical exploration (14 resection, seven inspection). Real time gray scale ultrasonography, color Doppler ultrasonography, computed tomography, and angiography were performed in all patients to evaluate portal vein invasion, and the images were compared with the histopathologic or surgical inspection findings. On comparison between gray scale ultrasonographic and color Doppler ultrasonographic images, the tumor-vessel relations were visualized more clearly on color Doppler ultrasonography than on gray scale ultrasonography in 14 (22.2%) of 63 vessels. The sensitivity of color Doppler ultrasonography, computed tomography, and angiography for diagnosing portal invasion was 73.7%, 73.7%, and 73.6%, and the specificity was 95.1%, 95.5%, and 90.9%, respectively; the overall accuracy was 84.1%, 88.9% and 85.7%, respectively. A mosaic signal pattern was observed in 12 vessels and showed an accuracy of 86.4% for diagnosing portal vein invasion. In conclusion, compared with gray scale ultrasonography, color Doppler ultrasonography provided improved images of the tumor and portal vein. Furthermore, the accuracy of color Doppler evaluation of portal vein invasion appears to be equal to that of computed tomography and angiography. Therefore, color Doppler ultrasonography may play an important role as an initial examination for evaluating portal vein invasion. PMID- 9401998 TI - Effective ultrasonographically guided intervention for diagnosis of musculoskeletal lesions. AB - Current algorithms recommend computed tomography or fluoroscopic guidance rather than ultrasonography for musculoskeletal intervention. We analyzed our ultrasonographically guided experience to evaluate its efficacy. Forty-seven patients underwent needle aspirates or biopsies or both in 13 extremity and 34 axial locations for 12 inflammatory lesions, 23 soft tissue masses, and 12 lesions arising from bone. Four lesions were initially imaged by ultrasonography; the remaining lesions were identified by computed tomography (25) or magnetic resonance imaging (18). Forty-six samples were diagnostic; one needle aspirate of an inflammatory mass yielded no diagnostic material. No complications occurred. Ultrasonographically guided musculoskeletal aspiration and biopsy are diagnostic and effective throughout the body, and with appropriate lesion access, they should be considered as an alternative to computed tomographic-guided procedures. PMID- 9401999 TI - Sonographic evolution of a living cervical pregnancy treated with intraamniotic instillation of methotrexate. PMID- 9402000 TI - Pseudoaneurysm formation in the breast after core needle biopsy. PMID- 9402001 TI - Iatrogenic creation of a monoamniotic twin gestation in severe twin-twin transfusion syndrome. PMID- 9402002 TI - Integrin signaling. AB - Integrins provide dynamic links between cells and extracellular matrix molecules. Although integrins were originally viewed as relatively simple adhesion molecules, it soon became clear that intracellular signal transduction initiated by integrins is centrally involved in many cellular processes. In fact, a remarkable number of classical signaling pathways are now known to be activated or modified by the interactions of cells with matrix proteins via integrins. These integrin signaling responses can also involve many other extracellular and intracellular molecules. The following mini-reviews were solicited from some of the future leaders in the field of integrin signaling. They examine selected important portions of this field, provide conceptual syntheses from a large and confusing literature, and then propose novel testable ideas. These ideas should encourage dialogue and open new avenues of research in this rapidly expanding, exciting field. PMID- 9402003 TI - Structural basis of integrin-mediated signal transduction. AB - Integrins are a family of alpha/beta heterodimers of cell adhesion receptors that mediate cell-extracellular matrix and cell-cell interactions. Both alpha and beta subunits have a large extracellular domain and a short cytoplasmic domain. The alpha subunit has seven sequence repeats of 60-70 residues in its N-terminal region. The beta-propeller model, in which seven four-stranded beta-sheets are arranged in a torus around a pseudosymmetry axis, has been proposed as a structural model of these seven repeats. Several predicted loops critical for ligand binding have been identified in the upper face of the proposed beta propeller model. Several alpha subunits (e.g., alpha 2, alpha L and alpha M) have I-domains of about 200 residues inserted between their second and third repeats. These I-domains adopt a Rossman-fold structure and have major ligand and cation binding sites (the MIDAS site) on their surfaces. The beta subunit has an I domain-like structure in its N-terminal region. This structure includes multiple sequences/conserved oxygenated residues critical for ligand binding (e.g., Asp 119 in beta 3), and non-conserved residues critical for ligand specificities. Several "activation-dependent" epitopes have been identified in the Cys-rich (stalk) region of beta 1. It has yet to be determined how these multiple ligand binding sites in the alpha and beta subunits are involved in ligand binding, and how conformational changes on activation/ligand occupancy relate to signal transduction. PMID- 9402005 TI - Integrin signaling: building connections beyond the focal contact? AB - The integrin family of adhesion receptors is notable for its bi-directional signaling properties, changing extracellular conformation and ligand binding affinity in response to external agonists, and inducing complex intracellular events following ligation. The interaction of integrins with intracellular or transmembrane moieties has been extensively studied in order to define the mechanisms of bi-directional signaling. In particular, the recruitment of integrins to focal contacts puts them in proximity with many cytoskeletal and signaling molecules. The importance of these structures in connecting integrins and signaling pathways has been well described. However, the purpose of this minireview is to explore additional protein interactions and how these might serve as an alternative means in connecting integrins and signaling pathways. PMID- 9402004 TI - Integrin cytoplasmic domains as connectors to the cell's signal transduction apparatus. AB - Integrins mediate the bidirectional transfer of signals across the plasma membrane. Integrin cytoplasmic domains provide one pathway linking integrin engagement with the cell's signal transduction apparatus. Recent structure function studies have defined regions of beta cytoplasmic domains required for integrin function and have identified distinct roles for individual alpha cytoplasmic domains in regulating cell behavior. Newly identified proteins that bind to integrin alpha and beta cytoplasmic domains have provided new insights and new questions into the mechanisms involved in integrin signaling. PMID- 9402007 TI - Integrin receptor signaling: the propagation of an alpha-helix model. AB - Upon ligand binding to integrin receptors, a transmembrane conformation change occurs, which is required for the engagement of the actin cytoskeleton. Integrin receptor latency clearly involves the proximal portions of the alpha and beta cytoplasmic domains. Several experiments suggest that these two regions, which are highly conserved among integrins, may be associated, and this association is the structural basis for latency. We propose that ligand binding leads to a disruption of this association, which allows for the folding of the proximal beta cytoplasmic domain. Thus, in this model, the alpha chain association keeps the beta unfolded, and ligand binding leads to the propagation of an alpha helix from the transmembrane domain through the proximal beta cytoplasmic domain, leading to signal transduction. PMID- 9402006 TI - Allostery and proteolysis: two novel modes of regulating integrin function. AB - Integrins are involved in transmitting signals between the cytoplasm and the extracellular matrix. Importantly, the transfer of information is bi-directional: signals flow inside-out and outside-in. Here, I discuss two potential modes by which integrin function is likely to be regulated. It is hypothesized that the integrin cytoplasmic tails are proteolytic substrates, and that cleavage of the cytoplasmic domain regulates the ligand binding affinity of integrins. It is also hypothesized that the ligand binding site is allosterically regulated by separate divalent ion binding sites that independently control ligand association and dissociation rate. Both hypotheses are suggested by reports in the literature and can be tested experimentally. PMID- 9402008 TI - Alternatively spliced variants: a new view of the integrin cytoplasmic domain. AB - A large number of studies have underscored a major role for the integrin alpha beta cytoplasmic domains in the modulation of cell functions. Cytoplasmic domain variants of the beta 1, beta 3, beta 4, alpha 3, alpha 6 and alpha 7 subunits have been described. These molecules are generated by alternative splicing events and are expressed in a cell- or tissue-type specific manner. Some of these variants (beta 1C, beta 1D, alpha 6A and alpha 7A) are predominantly expressed upon differentiation and have been shown to be regulated during development. The studies on the structure-function relationship of the integrin variant subunits, published between 1989 and now, will be reviewed here for the first time. The results demonstrate that differences in the cytoplasmic domain do not affect either the alpha beta heterodimer formation or the ligand specificity. Instead, alternatively spliced integrin cytoplasmic domains appear to be essential modulators of receptor localization, cell proliferation and migration, as well as phosphorylation of signaling molecules. These observations lead to the current hypothesis that cell-type specific regulation of alternatively spliced integrin cytoplasmic domains may provide a highly specialized mechanism to control cell growth and intracellular signaling pathways. PMID- 9402009 TI - Focal adhesion kinase in integrin signaling. AB - Focal adhesion kinase (FAK) has recently been established as a key component of the signal transduction pathways triggered by integrins. Aggregation of integrins and the cytoskeletal proteins tensin, paxillin and talin is proposed to be responsible for FAK activation and autophosphorylation by integrins in cell adhesion. Activation and autophosphorylation of FAK lead to its binding to a number of intracellular signaling molecules, including Src, Grb2 and PI 3-kinase. FAK/Src association activates both kinases, which act on the potential substrates tensin, paxillin and p130cas. Besides cytoskeletal regulation, FAK phosphorylation of paxillin and p130cas could also lead to MAP kinase pathway by adaptor proteins such as Crk and Nck. Recent studies indicated that integrin signaling through FAK causes increased cell migration and potentially regulates cell proliferation and survival. Future challenges will include clarifying the roles of signaling pathways downstream of FAK in cell migration and cell cycle regulation. PMID- 9402010 TI - Synthesis and conformational properties of a recombinant C-propeptide of human type III procollagen. AB - A cDNA was prepared that coded for the signal peptide of type III procollagen linked to the complete C-propeptide of the protein. The cDNA was then used to express the protein in a baculovirus recombinant system. Recombinant protein was recovered as a trimer from the medium of transfected cells in a yield of 1 to 2.5 mg per liter. Mapping of peptide fragments with and without reduction indicated that the protein contained the expected interchain disulfide bonds. Analysis by circular dichroism suggested that the conformation of the protein corresponded to the native conformation. Therefore, the protein should be appropriate for further tests of its biological function and analysis of structure by X-ray diffraction. PMID- 9402011 TI - Non-glycosaminoglycan bearing domains of perlecan and aggrecan influence the utilization of sites for heparan and chondroitin sulfate synthesis. AB - Perlecan and aggrecan are proteoglycans that receive primarily heparan sulfate and chondroitin sulfate side chains, respectively. Their large multidomained core proteins have little or no homology to each other and their glycosaminoglycan (GAG) attachment sites are restricted to certain domains only. We examined the involvement of the non-GAG bearing domains in designating the GAG type added to the GAG attachment domain by preparing cDNA constructs that expressed perlecan/aggrecan chimeras as recombinant products in COS-7 cells and then determining the size and GAG composition of the recombinant products. The results showed that domain I of perlecan receives primarily (73-81%) heparan sulfate when coupled with domain II and III of perlecan, but when coupled with the G3 domain of aggrecan, it receives primarily (59-63%) chondroitin sulfate. Furthermore, the chondroitin sulfate attachment region of aggrecan received GAG side chains more readily when coupled to the G3 domain of aggrecan than when coupled to domains II and III of perlecan. The GAG side chains on all these recombinant products were small and similar in size. These findings indicate that the utilization of attachment sites for heparan and chondroitin sulfate or the sulfation of these GAGs can be influenced, in part, by non-GAG bearing domains. PMID- 9402012 TI - Differentiation-dependent expression of laminin-8 (alpha 4 beta 1 gamma 1) mRNAs in mouse 3T3-L1 adipocytes. AB - We report that laminin-8 (alpha 4 beta 1 gamma 1) is the specific isoform of laminin synthesized in adipocytes. Reverse transcription-polymerase chain reaction (RT-PCR) of mRNA from mouse 3T3-L1 cells with paired primers for alpha 1, alpha 2, alpha 3, alpha 4, alpha 5, beta 1, beta 2, beta 3, gamma 1 and gamma 2 laminins yielded amplified fragments only for alpha 4, beta 1 and gamma 1. A polyclonal antibody against mouse laminin-1 (alpha 1 beta 1 gamma 1) precipitated alpha 4 in addition to beta 1 and gamma 1, while the antibody against a deduced peptide sequence of mouse alpha 4 in addition to beta 1 and gamma 1 in addition to alpha 4. Thus, laminin-8 (alpha 4 beta 1 gamma 1) is the only isoform expressed in 3T3-L1 cells. Northern blots showed that the levels of alpha 4, beta 1 and gamma 1 mRNAs increased 2.5-fold during adipose conversion of 3T3-L1 cells. A 1062 bp cDNA fragment cloned by RT-PCR demonstrated a polymorphism in the mouse alpha 4 gene which would lead to five amino acid changes in the domain G. PMID- 9402013 TI - The Escherichia coli genome sequence: the end of an era or the start of the FUN? AB - Our dream of determining the entire Escherichia coli K12 genome sequence has been realized. This calls for new approaches for the analysis of gene expression and function in biology's best-understood organism. Comparison of the E. coli genome sequence with others will provide important taxonomic insights and have implications for the study of bacterial virulence. Approximately 20% of E. coli genes have been designated FUN genes, because they have no known function or homologies to sequence databases. FUN genes promise to have an exciting impact on bacterial research. The post-genome era requires novel strategies that address gene regulation at the level of the entire cell. These strategies need to supersede the reductionist approach to genetic analysis. Only then will the genome sequence lead us to an understanding of how a bacterial cell really works. PMID- 9402014 TI - The non-standard genetic code of Candida spp.: an evolving genetic code or a novel mechanism for adaptation? AB - A number of yeasts of the genus Candida translate the standard leucine-CUG codon as serine. This unique genetic code change is the only known alteration to the universal genetic code in cytoplasmic mRNAs, of either eukaryotes or prokaryotes, which involves reassignment of a sense codon. Translation of CUG as serine in these species is mediated by a novel serine-tRNA (ser-tRNACAG), which uniquely has a guanosine at position 33, 5' to the anticodon, a position that is almost invariably occupied by a pyrimidine (uridine in general) in all other tRNAs. We propose that G-33 has two important functions: lowering the decoding efficiency of the ser-tRNACAG and preventing binding of the leucyl-tRNA synthetase. This implicates this nucleotide as a key player in the evolutionary reassignment of the CUG codon. In addition, the novel ser-tRNACAG has 1-methylguanosine (m1G-37) at position 37, 3' to the anticodon, which is characteristic of leucine, but not serine tRNAs. Remarkably, m1G-37 causes leucylation of the ser-tRNACAG both in vitro and in vivo, making the CUG codon an ambiguous codon: the polysemous codon. This indicates that some Candida species tolerate ambiguous decoding and suggests either that (i) the genetic code change has not yet been fully established and is evolving at different rates in different Candida species; or (ii) CUG ambiguity is advantageous and represents the final stage of the reassignment. We propose that such dual specificity indicates that reassignment of the CUG codon evolved through a mechanism that required codon ambiguity and that ambiguous decoding evolved to generate genetic diversity and allow for rapid adaptation to environmental challenges. PMID- 9402015 TI - Molecular mechanisms of Campylobacter fetus surface layer protein expression. AB - Cells of the Gram-negative bacteria Campylobacter fetus are covered by monomolecular arrays of surface layer proteins (SLPs) critical for both persistence in their natural hosts and for virulence. For C. fetus cells, expression of SLPs essentially eliminates C3b binding and their antigenic variation thwarts host immunological defences. Each cell possesses multiple partially homologous and highly conserved SLP gene cassettes, tightly clustered in the genome, that encode SLPs of 97-149 kDa. These attach non-covalently via a conserved N-terminus to the cell wall lipopolysaccharide. Recent studies indicate that C. fetus reassorts a single promoter, controlling SLP expression, and one, or more, complete open reading frame strictly by DNA inversion, and that rearrangement is independent of the distance between sites of inversion. In contrast to previously reported programmed DNA inversion systems, inversion in C. fetus is recA-dependent. These rearrangements permit variation in protein expression from the family of SLP genes and suggest an expanding paradigm of programmed DNA rearrangements among microorganisms. PMID- 9402016 TI - Non-palindromic attl sites of integrons are capable of site-specific recombination with one another and with secondary targets. AB - Genes borne on cassettes are mobile owing to site-specific recombination systems called integrons, which have created various combinations of antibiotic resistance genes in R-plasmids. In these processes, the palindromic site, attC (59-base element), at cassette junctions has been proposed as being essential. Excised and circularized cassettes have been found to integrate with preference for an attl site at one end of the conserved sequence in integrons. In this work, we give evidence that recombination is possible in the absence of the highly organized attC sites between the more simply organized attl sites. Furthermore, at a very low frequency representing the background in our recombination assay, we observed cross-overs between attl and secondary sites. To characterize recombination excluding the attC sites, we have used naturally occurring attl variants and constructed mutants. The cross-over point was identified between a guanine and a thymine in attl using point mutations. Progressive deletions showed the extent of attl and identified two important regions in the conserved sequence 5' of the cross-over point. A region 27-36 bp 5' of attl influenced recombination with attC sites only, whereas a sequence 9-14 bp 5' of the cross-over point in attl was important for recombination with both attl and attC. Recombination between attl and secondary sites could allow fusion of the conserved sequence encoding the integron site-specific recombinase to new sequences. PMID- 9402017 TI - The finM promoter and the traM promoter are the principal promoters of the traM gene of the antibiotic resistance plasmid R100. AB - finP multicopy repression and traJ multicopy derepression indicate that the ratio of sense to antisense transcripts is important in the regulation of R100 conjugation. The extension of R100 traM transcripts into traJ shows that promoters in traM can affect this ratio, making the regulation of traM transcription important in the regulation of R100 conjugation. Since R100 traM, traY and tral proteins bind to the traM promoter region, we examined traM transcription in R100-1 traM, traY and tral mutants and compared it with traM transcription in both R100-1 and R100. We verified that the traM and finM promoters provide virtually all the transcripts originating in the R100-1 traM gene. When either is deleted, as in VAR22 or VAR30, the remaining promoter is highly active. We show here that traY positively regulates R100-1 traM transcription, as has been found for F. We found that tral did not regulate R100 1 traM transcription. The measured activity of the native R100 traM promoter was 12% of that in R100-1, whereas the native R100 finM promoter was 45% of that in R100-1. These data and data from the R100-1 traY and tral mutants show that the activities of the two promoters varied independently. PMID- 9402018 TI - The role of ribosomal RNAs in macrolide resistance. AB - Macrolides are bacteriostatic antibiotics which interfere with the peptidyltransfer function of the ribosome. We have investigated the molecular mechanisms underlying macrolide resistance in Mycobacterium smegmatis, an eubacterium carrying two rRNA operons. Surprisingly, drug resistance was associated not with alterations in ribosomal proteins, but with a single point mutation in the peptidyltransferase region of one of the two 23S RNA genes, i.e. A2058-->G or A2059-->G. This mutation resulted in a heterozygous organism with a mutated and a wild-type rRNA operon respectively. Reverse transcriptase sequencing indicated the expression of both wild-type and mutated rRNAs. The mutated operon was introduced into genetically engineered rrn- strains of M. smegmatis carrying a single functional rRNA operon and into parental M. smegmatis with two chromosomal rRNA operons, using gene transfer as well as gene replacement techniques. The results obtained demonstrate the dominant nature of resistance. As exemplified in our results on macrolide resistance, a complete set of genetic tools is now available, which allows questions of dominance vs. recessivity and gene dosage effects in eubacterial ribosomal nucleic acids to be addressed experimentally in vivo. PMID- 9402019 TI - Cardiolipin is not essential for the growth of Saccharomyces cerevisiae on fermentable or non-fermentable carbon sources. AB - Cardiolipin is a unique dimeric phospholipid, which is present throughout the eukaryotic kingdom and is specifically localized in mitochondrial membranes. It is widely believed that mitochondria possess an essential requirement for this phospholipid. To determine whether cardiolipin is essential for yeast growth, we generated a cardiolipin synthase null mutant by disrupting the CLS1 gene (open reading frame YDL142c on chromosome IV) of Saccharomyces cerevisiae. Biochemical analysis of the mutant indicated that it had no cardiolipin synthase activity and no cardiolipin in its membranes. The enzyme phosphatidylglycerolphosphate synthase, which catalyses the committed step of the cardiolipin pathway, remained unaffected in the null mutant. Haploid cells containing the null allele are viable in media containing glucose, galactose or glycerol/ethanol as the sole carbon source, although growth in galactose or glycerol/ethanol is somewhat reduced in the mutant compared with the wild type. These results indicate that cardiolipin is not essential for the growth of S. cerevisiae in fermentable or non-fermentable carbon sources. PMID- 9402020 TI - Regulation of plasmid R1 replication: PcnB and RNase E expedite the decay of the antisense RNA, CopA. AB - The replication frequency of plasmid R1 is controlled by an unstable antisense RNA, CopA, which, by binding to its complementary target, blocks translation of the replication rate-limiting protein RepA. Since the degree of inhibition is directly correlated with the intracellular concentration of CopA, factors affecting CopA turnover can also alter plasmid copy number. We show here that PcnB (PAPl-a poly(A)polymerase of Escherichia coli) is such a factor. Previous studies have shown that the copy number of ColE1 is decreased in pcnB mutant strains because the stability of the RNase E processed form of RNAI, the antisense RNA regulator of ColE1 replication, is increased. We find that, analogously, the twofold reduction in R1 copy number caused by a pcnB lesion is associated with a corresponding increase in the stability of the RNase E generated 3' cleavage product of CopA. These results suggest that CopA decay is initiated by RNase E cleavage and that PcnB is involved in the subsequent rapid decay of the 3' CopA stem-loop segment. We also find that, as predicted, under conditions in which CopA synthesis is unaffected, pcnB mutation reduces RepA translation and increases CopA stability to the same extent. PMID- 9402021 TI - Structural determinants of processing and secretion of the Haemophilus influenzae hap protein. AB - Haemophilus influenzae elaborates a surface protein called Hap, which is associated with the capacity for intimate interaction with cultured epithelial cells. Expression of hap results in the production of three protein species: outer membrane proteins of approximately 155 kDa and 45 kDa and an extracellular protein of approximately 110 kDa. The 155 kDa protein corresponds to full-length mature Hap (without the signal sequence), and the 110 kDa extracellular protein represents the N-terminal portion of mature Hap (designated Haps). In the present study, we examined the mechanism of processing and secretion of Hap. Site directed mutagenesis suggested that Hap is a serine protease that undergoes autoproteolytic cleavage to generate the 110 kDa extracellular protein and the 45 kDa outer membrane protein. Biochemical analysis confirmed this conclusion and established that cleavage occurs on the bacterial cell surface. Determination of N-terminal amino acid sequence and mutagenesis studies revealed that the 45 kDa protein corresponds to the C-terminal portion of Hap, starting at N1037. Analysis of the secondary structure of this protein (designated Hap beta) predicted formation of a beta-barrel with an N-terminal transmembrane alpha-helix followed by 14 transmembrane beta-strands. Additional analysis revealed that the final beta-strand contains an amino acid motif common to other beta-barrel outer membrane proteins. Upon deletion of this entire C-terminal consensus motif, Hap could no longer be detected in the outer membrane, and secretion of Haps was abolished. Deletion or complete alteration of the final three amino acid residues had a similar but less dramatic effect, suggesting that this terminal tripeptide is particularly important for outer membrane localization and/or stability of the protein. In contrast, isolated point mutations that disrupted the amphipathic nature of the consensus motif or eliminated the C-terminal tryptophan had no effect on outer membrane localization of Hap or secretion of Haps. These results provide insight into a growing family of Gram-negative bacterial exoproteins that are secreted by an IgA1 protease-like mechanism; in addition, they contribute to a better understanding of the structural determinants of targeting of beta-barrel proteins to the bacterial outer membrane. PMID- 9402022 TI - FIS modulates growth phase-dependent topological transitions of DNA in Escherichia coli. AB - The Escherichia coli DNA-binding protein FIS serves as a DNA architectural factor in two unrelated enzymatic reactions, the site-specific inversion of DNA and transcriptional activation of stable RNA promoters. In both these processes, FIS facilitates the assembly and dynamic transitions of two structurally distinct nucleoprotein complexes. We have proposed previously that, in these systems, FIS stabilizes writhed DNA microloops by binding at multiple helically phased sites in DNA. However, FIS also binds and bends DNA at many non-specific sites and, at its maximum levels in the early exponential phase, FIS could potentially occupy a considerable part of the E. coli chromosome. Here, we show that fis affects growth phase-specific alterations in the supercoiling level of DNA. Expression of fis accelerates the accumulation of moderately supercoiled plasmids in stationary phase, which are stabilized by FIS after nutritional shift-up. In accordance with such a function, FIS modulates the relaxing and supercoiling activities of topoisomerases in vitro in a way that keeps DNA in a moderately supercoiled state. Our results suggest that the primary role of FIS is to modulate chromosomal dynamics during bacterial growth. PMID- 9402023 TI - The rap and hor proteins of Erwinia, Serratia and Yersinia: a novel subgroup in a growing superfamily of proteins regulating diverse physiological processes in bacterial pathogens. AB - The enteric bacterium Serratia marcescens is an opportunistic human pathogen. The strain ATCC39006 makes the red pigment, prodigiosin (Pig), and the beta-lactam antibiotic carbapenem (Car). Mutants were isolated that were concomitantly defective for Pig and Car production. These mutants were found to have a mutation in the rap gene (Regulation of Antibiotic and Pigment). Sequence analysis of the rap gene revealed a predicted protein product showing strong homology to SlyA, originally thought to be a haemolytic virulence determinant in Salmonella typhimurium. Homologues of rap were detected in several bacterial genera, including Salmonella, Yersinia, Enterobacter, and species of the plant pathogen, Erwinia. The Erwinia hoeEr (homologue of rap) and the Yersinia horYe genes were also found to be very similar to rap and slyA. Marker exchange mutagenesis of horEr revealed that it encoded a regulatory protein controlling the production of antibiotic and exoenzyme virulence determinants in the phytopathogen, Erwinia carotovora subspecies carotovora. We have shown that these new homologues of SlyA form a highly conserved subgroup of a growing superfamily of bacterial regulatory proteins controlling diverse physiological processes in human, animal and plant pathogens. PMID- 9402024 TI - Analysis of the carbapenem gene cluster of Erwinia carotovora: definition of the antibiotic biosynthetic genes and evidence for a novel beta-lactam resistance mechanism. AB - Members of two genera of Gram-negative bacteria, Serratia and Erwinia, produce a beta-lactam antibiotic, 1-carbapen-2-em-3-carboxylic acid. We have reported previously the cloning and sequencing of the genes responsible for production of this carbapenem in Erwinia carotovora. These genes are organized as an operon, carA--H, and are controlled by a LuxR-type transcriptional activator, encoded by the linked carR gene. We report in this paper the genetic dissection of this putative operon to determine the function of each of the genes. We demonstrate by mutational analysis that the products of the first five genes of the operon are involved in the synthesis of the carbapenem molecule. Three of these, carABC, are absolutely required. In addition, we provide evidence for the existence of a novel carbapenem resistance mechanism, encoded by the CarF and carG genes. Both products of these overlapping and potentially translationally coupled genes have functional, N-terminal signal peptides. Removal of these genes from the Erwinia chromosome results in a carbapenem-sensitive phenotype. We assume that these novel beta-lactam resistance genes have evolved in concert with the biosynthetic genes to ensure 'self-resistance' in the Erwinia carbapenem producer. PMID- 9402025 TI - uvrD mutations enhance tandem repeat deletion in the Escherichia coli chromosome via SOS induction of the RecF recombination pathway. AB - It has previously been shown that recombination between tandem repeats is not significantly affected by a recA mutation in Escherichia coli. Here, we describe the activation of a RecA-dependent recombination pathway in a hyper-recombination mutant. In order to analyse how tandem repeat deletion may proceed, we searched for mutants that affect this process. Three hyper-recombination clones were characterized and shown to be mutated in the uvrD gene. Two of the mutations were identified as opal mutations at codons 130 and 438. A uvrD::Tn5 mutation was used to investigate the mechanism of deletion formation in these mutants. The uvrD mediated stimulation of deletion was abolished by a lexAind3 mutation or by inactivation of either the recA, recF, recQ or ruvA genes. We conclude that (i) this stimulation requires SOS induction and (ii) tandem repeat recombination in uvrD mutants occurs via the RecF pathway. In uvrD+ cells, constitutive expression of SOS genes is not sufficient to stimulate deletion formation. This suggests that the RecF recombination pathway activated by SOS induction is antagonized by the UvrD protein. Paradoxically, we observed that the overproduction of UvrD from a plasmid also stimulates tandem repeat deletion. However, this stimulation is RecA independent, as is deletion in a wild-type strain. We propose that the presence of an excess of the UvrD helicase favours replication slippage. This work suggests that the UvrD helicase controls a balance between different routes of tandem repeat deletion. PMID- 9402026 TI - Blocking rolling circle replication with a UV lesion creates a deletion hotspot. AB - UV light irradiation increases genetic instability by causing mutations and deletions. The mechanism of UV-induced rearrangements was investigated making use of deletion-prone plasmids. Chimeric plasmids carrying pBR322 and M13 replication origins undergo deletions that join the M13 replication origin to a random nucleotide. A restriction fragment was UV irradiated, introduced into such a hybrid plasmid and deletions formed at the M13 origin were analysed. In most of the deletant molecules, the M13 replication nick site was linked to a nucleotide in the irradiated fragment, showing that UV lesions are deletion hotspots. These deletions were independent of the UvrABC excision repair proteins, suggesting that the deletogenic structure is the lesion itself and not a repair intermediate. They were not found in the absence of M13 replication, indicating that they result from the encounter of the M13 replication fork with the UV lesion. Furthermore, UV-induced deletions occurred independently of pBR322 replication. We conclude that, in contrast to pBR322 replication forks, M13 replication forks blocked by UV lesions are deletion prone. We propose that the deletion-prone properties of a UV-arrested polymerase depend on the associated helicase. PMID- 9402027 TI - Telomeres of the linear chromosomes of Lyme disease spirochaetes: nucleotide sequence and possible exchange with linear plasmid telomeres. AB - Bacteria of the spirochaete genus Borrelia have linear chromosomes about 950 kbp in size. We report here that these linear chromosomes have covalently closed hairpin structures at their termini that are similar but not identical to those reported for linear plasmids carried by these organisms. Nucleotide sequence analysis of the chromosomal telomeric regions indicates that unique, apparently functional genes lie within a few hundred bp of each of the telomeres, and that there is an imperfect 26 bp inverted repeat at the two telomeres. In addition, we characterize a major chromosomal length polymorphism within the right telomeric regions of various Borrelia isolates, and show that sequences similar to those near the right telomere are often found on linear plasmids in B. burgdorferi (sensu stricto) isolates from nature. Sequences similar to a number of other regions of the chromosome, including those near the left telomere, were not found on B. burgdorferi plasmids. These observations suggest that there has been historical exchange of genetic information between the linear plasmids and the right end of the linear chromosome. PMID- 9402028 TI - Repair by recombination of DNA containing a palindromic sequence. AB - We report here that homologous recombination functions are required for the viability of Escherichia coli cells maintaining a 240 bp chromosomal inverted repeat (palindromic) sequence. Wild-type cells can successfully replicate this palindrome but recA, recB or recC mutants carrying the palindrome are unviable. The dependence on homologous recombination for cell viability is overcome in sbcC mutants. Directly repeated copies of the DNA containing the palindrome are rapidly resolved to single copies in wild-type cells but not in sbcC mutants. Our results suggest that double-strand breaks introduced at the palindromic DNA sequence by the SbcCD nuclease are repaired by homologous recombination. The repair is conservative and the palindrome is retained in the repaired chromosome. We conclude that SbcCD can attack secondary structures but that repair conserves the DNA sequence with the potential to fold. PMID- 9402029 TI - Mammalian heterotrimeric G-protein-like proteins in mycobacteria: implications for cell signalling and survival in eukaryotic host cells. AB - Mammalian heterotrimeric GTP-binding protein (G proteins) are involved in transmembrane signalling that couples a number of receptors to effectors mediating various physiological processes in mammalian cells. We demonstrate that bacterial proteins such as a Ras-like protein from Pseudomonas aeruginosa or a 65 kDa protein from Mycobacterium smegmatis can form complexes with human or yeast nucleoside diphosphate kinase (Ndk) to modulate their nucleoside triphosphate synthesizing specificity to GTP or UTP. In addition, we demonstrate that bacteria such as M. smegmatis or Mycobacterium tuberculosis harbour proteins that cross react with antibodies against the alpha-, beta- or the gamma-subunits of heterotrimeric G proteins. Such antibodies also after the GTP synthesizing ability of specific membrane fractions isolated from glycerol gradients of such cells, suggesting that a membrane-associated Ndk-G-protein homologue complex is responsible for part of GTP synthesis in these bacteria. Indeed, purified Ndk from human erythrocytes and M. tuberculosis showed extensive complex formation with the purified mammalian alpha- and beta-G-protein subunits and allowed specific GTP synthesis, suggesting that such complexes may participate in transmembrane signalling in the eukaryotic host. We have purified the alpha-, beta- and gamma-subunit homologues from M. tuberculosis and we present their internal amino acid sequences as well as their putative homologies with mammalian subunits and the localization of their genes on the M. tuberculosis genome. Using oligonucleotide probes from the conserved regions of the alpha- and gamma-subunit of M. tuberculosis G-protein homologue, we demonstrate hybridization of these probes with the genomic digest of M. tuberculosis H37Rv but not with that of M. smegmatis, suggesting that M. smegmatis might lack the genes present in M. tuberculosis H37Rv. Interestingly, the avirulent strain H37Ra showed weak hybridization with these two probes, suggesting that these genes might have been deleted in the avirulent strain or are present in limited copy numbers as opposed to those in the virulent strain H37Rv. PMID- 9402030 TI - Structural principles of prokaryotic gene regulatory proteins and the evolution of repressors and gene activators. PMID- 9402031 TI - Effects of thalidomide on neutrophil respiratory burst, chemotaxis, and transmigration of cytokine- and endotoxin-activated endothelium. AB - Vascular endothelium activated by endotoxin and cytokines plays an important role in organ inflammation and blood leukocyte recruitment. Neutrophils, which are a homogeneous population of effector cells, are rapidly attracted in large numbers to sites of inflammation where they form an early response to infection or injury. Excessive production of various interleukins, TNF, arachidonic acid metabolites, and other substances by neutrophils and macrophages results in systemic endothelial cell injury, a fundamental problem. In the present study, we investigated in vitro the effects of thalidomide (THD) on activation of endothelial cells for enhanced transmigration of neutrophils by lipopolysaccharide (LPS), tumor necrosis factor-alpha (TNF), and interleukin-1 (IL-1). Modulation of endotoxin- and cytokine-induced neutrophil chemotaxis and respiratory burst by THD were also studied. Treatment of HUVEC with THD in combination with LPS, TNF, and IL-1, respectively, antagonized LPS-activated transmigration of neutrophils but stimulated the effects of TNF and IL-1. All of the agents used-THD, LPS, TNF, and IL-1-inhibited neutrophil chemotaxis. Addition of THD to the neutrophils had no effect on LPS-inhibited chemotaxis whereas the TNF- and IL-1-induced chemotaxis was modulated in a bimodal manner. However, THD failed to influence neutrophil respiratory burst activity. Results demonstrate that THD differentially affects mediator-induced activation of HUVEC and neutrophils. PMID- 9402032 TI - Multiple calcium channels are required for pituitary adenylate cyclase-activating polypeptide-induced catecholamine secretion from bovine cultured adrenal chromaffin cells. AB - The effects of L-, N-, P- and Q-type calcium channel antagonists and (+/-)-BayK 8644 on catecholamine release induced by pituitary adenylate cyclase-activating polypeptide (PACAP-27) were investigated in bovine cultured adrenal chromaffin cells. PACAP-27 induced the release of 4-15% of the total cellular catecholamines over 7 min, with an EC50 of 20 nM and the effect approaching maximum at 100 nM. Catecholamine release was fully dependent on the presence of extracellular calcium. The dihydropyridine nitrendipine which inhibits L-type calcium channels inhibited PACAP-27-induced secretion in a concentration dependent manner with an inhibition of 20-30% at 1 microM. In contrast, (+/-)-BayK-8644, which prolongs the opening of L-type calcium channels produced a concentration-dependent increase in PACAP-27-induced catecholamine release with 1 microM increasing release by 40-60%. Blockade of N-type calcium channels with omega-conotoxin GVIA reduced release by 5-15%. Block of P-type channels with low concentrations of omega-agatoxin IVA (< or = 30 nM) had no significant effect on release, while higher concentrations (100-300 nM) which block Q-type channels reduced release by up to 15%. omega-Conotoxin MVIIC, an antagonist of Q-type calcium channels and also of N- and P-type channels, inhibited release in a concentration-dependent manner with a near maximum effect of 30-50% produced by 300 nM. The combination of omega-conotoxin GVIA and omega-agatoxin IVA reduced release by 40-50%. Addition of omega-conotoxin MVIIC (300 nM) to the combination of omega-conotoxin GVIA (10 nM) and omega-agatoxin IVA (100 nM) did not inhibit catecholamine release more than with omega-conotoxin GVIA and omega-agatoxin IVA alone, indicating that 100 nM omega-agatoxin IVA was sufficient to block the Q-type calcium channels. When nitrendipine was used together with omega-conotoxin GVIA, omega-agatoxin IVA and omega-conotoxin MVIIC, catecholamine release induced by 20 nM or 100 nM PACAP-27 was reduced by 70-85%. Taken together these results suggest that influx of calcium through multiple different voltage-sensitive calcium channels mediate PACAP-27-induced catecholamine release from bovine chromaffin cells, and that L-, N- and Q-channels contribute to this response. PMID- 9402033 TI - Imidazole antimycotics affect the activity of various ion channels and insulin secretion in mouse pancreatic B-cells. AB - In the present paper the effects of antimycotics with imidazole structure on the activity of various ion currents of mouse pancreatic B-cells and insulin secretion from isolated islets have been studied. Clotrimazole (0.1-10 microM, bath solution without albumin) reversibly inhibited the whole-cell K + ATP current studied with the patch-clamp technique and concomitantly depolarized the membrane potential. Two other structurally related compounds, econazole and ketoconazole, exhibited similar effects on the whole-cell K + ATP current. Clotrimazole also inhibited the current through single K + ATP channels measured in the inside-out configuration. According to these results it seems unlikely that a cytoplasmic factor is involved in the action of clotrimazole on K + ATP currents. Clotrimazole (10 microM) also reduced the current through voltage dependent Ca2+ and K+ channels and altered inactivation kinetics. Moreover, clotrimazole reversibly abolished a recently described inward current which is induced by hypotonic cell swelling. The results show that clotrimazole altered the activity of all ion currents in B-cells investigated in this study. Clotrimazole (3-100 microM, solution with albumin) irreversibly inhibited insulin secretion from isolated islets. With econazole and ketoconazole similar effects on hormone release were observed. The changes in the activity and kinetics of voltage-dependent Ca2+ and K+ currents are likely to contribute to the observed inhibition of insulin secretion. However, we cannot entirely rule out that imidazole antimycotics also interfere with a step in stimulus-secretion coupling distal to changes in membrane potential. PMID- 9402034 TI - Stimulated [35S]GTP gamma S binding by 5-HT1A receptor agonists in recombinant cell lines. Modulation of apparent efficacy by G-protein activation state. AB - G-protein activation by different 5-HT receptor ligands was investigated in h5 HT1A receptor-transfected C6-glial and HeLa cells using agonist-stimulated [35S] GTP gamma S binding to membranes in the presence of excess GDP. 5-HT (10 microM) stimulated [35S]GTP gamma S binding in the C6-glial membrane preparation to a larger extent than in the HeLa preparation; maximal responses with 30 microM GDP were 490 +/- 99 and 68 +/- 12%, respectively. With the 5-HT receptor agonists that were being investigated, the two preparations displayed the same rank order of potency for stimulation of [35S]GTP gamma S binding. In the C6-glial preparation at 0.3 microM GDP, the rank order of maximal effects was: 5-HT (1.00) > 8-OH-DPAT (0.90) = R(+)-8-OH-DPAT (0.87) = 5-CT (0.86) = L694247 (0.84) > S(-)8 OH-DPAT (0.68) = buspirone (0.67) = spiroxatrine (0.67) = flesinoxan (0.64) > ipsapirone (0.53) = (-)-pindolol (0.50) > SDZ216525 (0.25). However, differences in maximal response in the C6-glial preparation were magnified by increasing the GDP concentrations, indicating that the activity state of G-proteins can affect the maximal response. With the exception of 5-CT and L694247, increasing the amount of GDP to 30 microM and higher concentrations resulted in an attenuation of both the ligand's maximal effect (24 to 56%) and apparent potency (6 to 24 fold). Each of the [35S]GTP gamma S binding responses was mediated by a 5-HT1A receptor as indicated by the competitive blockade by WAY100635 and spiperone. Only 5-CT and L694247 in some conditions displayed an efficacy similar to that of 5-HT at the h5-HT1A receptor; the other agents with intrinsic activity are partial agonists at this receptor. The data also suggest that the activity state of the G-proteins is involved in the maximal effects that can be produced by activating the h5-HT1A receptor. PMID- 9402035 TI - Chronic administration of the selective dopamine uptake inhibitor GBR 12,909, but not cocaine, produces marked decreases in dopamine transporter density. AB - The chronic continuous infusion of cocaine produces partial behavioral tolerance to cocaine and tolerance to the inhibition of dopamine uptake by cocaine, without changing dopamine transporter binding. In order to examine more closely the dopaminergic contribution to this effect, the selective dopamine uptake inhibitor GBR 12,909 (30 mg/kg/day), cocaine (50 mg/kg/day), or vehicle, were continuously infused via osmotic minipump, and their effects on the dopamine transporter examined. Drug and vehicle pumps were implanted into male Sprague-Dawley rats and removed after seven days. [3H]WIN 35,428 binding and [3H]dopamine uptake were measured in caudate putamen and nucleus accumbens at varying intervals after pump removal. The Bmax for [3H]WIN 35,428 binding was decreased by approximately 75% in the caudate putamen and by 40% in the nucleus accumbens of GBR 12,909-treated rats both 1 and 4 days after pump removal, and was still significantly decreased after 10 days, but had returned to normal by 20 days post-treatment. In contrast, cocaine did not significantly alter [3H]WIN 35,428 binding. GBR 12,909 produced both tolerance to the inhibition of [3H]dopamine uptake by cocaine, and a decrease in total uptake of dopamine, in the caudate putamen, with no change in the nucleus accumbens. The persistent reduction of [3H]WIN 35,428 binding following continuous GBR 12,909 does not appear to result from residual drug binding. These findings suggest that GBR 12,909 and cocaine may bind to and regulate the dopamine transporter in different ways. PMID- 9402036 TI - Evaluation of the effects of a specific alpha 2-adrenoceptor antagonist, atipamezole, on alpha 1- and alpha 2-adrenoceptor subtype binding, brain neurochemistry and behaviour in comparison with yohimbine. AB - In the present study we evaluated the alpha 1- and alpha 2-adrenoceptor subtype binding, central alpha 2-adrenoceptor antagonist potency, as well as effects on brain neurochemistry and behavioural pharmacology of two alpha 2-adrenoceptor antagonists, atipamezole and yohimbine. Atipamezole had higher selectivity for alpha 2- vs. alpha 1-adrenoceptors than yohimbine regardless of the subtypes studied. Both compounds had comparable affinity for the alpha 2A-, alpha 2C- and alpha 2B-adrenoceptors, but yohimbine had significantly lower affinity for the alpha 2D-subtype. This may account for the fact that significantly higher doses of yohimbine than atipamezole were needed for reversal of alpha 2-agonist (medetomidine)-induced effects in rats (mydriasis) and mice (sedation and hypothermia). The effect on central monoaminergic activity was estimated by measuring the concentrations of transmitters and their main metabolites in whole brain homogenate. At equally effective alpha 2-antagonising doses in the rat mydriasis model, both drugs stimulated central noradrenaline turnover (as reflected by increase in metabolite levels) to the same extent. Atipamezole increased dopaminergic activity only slightly, whereas yohimbine elevated central dopamine but decreased central 5-hydroxytryptamine turnover rates. In behavioural tests, atipamezole (0.1-10 mg/kg) did not affect motor activity but stimulated food rewarded operant (FR-10) responding (0.03-3 mg/kg) whereas yohimbine both stimulated (1 mg/kg) and decreased (> or = 3 mg/kg) behaviour in a narrow dose range in these tests. In the staircase test, both antagonists increased neophobia, but in the two compartment test only yohimbine (> or = 3 mg/kg) decreased exploratory behaviour. The dissimilar effects of the antagonists on neurochemistry and behaviour are thought to be caused by non alpha 2-adrenoceptor properties of yohimbine. In conclusion, the alpha 2-antagonist atipamezole blocked all alpha 2-adrenoceptor subtypes at low doses, stimulated central noradrenergic activity and had only slight effects on behaviour under familiar conditions, but increased neophobia. The low affinity for the alpha 2D adrenoceptor combined with its unspecific effects complicates the use of yohimbine as pharmacological tool to study alpha 2-adrenoceptor physiology and pharmacology. PMID- 9402037 TI - Cannabinoid CB1 receptor-mediated inhibition of noradrenaline release in the human and guinea-pig hippocampus. AB - We examined the question of whether cannabinoid receptors modulating noradrenaline release are detectable in the brain of humans and experimental animals. For this purpose, hippocampal slices from humans, guinea-pigs, rats and mice and cerebellar, cerebrocortical and hypothalamic slices from guinea-pigs were incubated with [3H]noradrenaline and then superfused. Tritium overflow was evoked either electrically (0.3 or 1 Hz) or by introduction of Ca2+ ions (1.3 mM) [corrected] into Ca(2+)-free, K(+)-rich medium (25 mM) [corrected] containing tetrodotoxin 1 microM. Furthermore, the cAMP accumulation stimulated by forskolin 10 microM was determined in guinea-pig hippocampal membranes. We used the following drugs: the cannabinoid receptor agonists (-)-cis-3-[2-hydroxy-4-(1,1- dimethylheptyl)phenyl]-trans-4-(3-hydroxypropyl)cyclo-hexanol (CP-55,940) and R(+)-[2,3-dihydro-5-methyl-3- [(morpholinyl)methyl]pyrrolo[1,2,3-de]-1,4 benzoxazinyl]- (1-naphthalenyl)methanone (WIN 55,212-2), the inactive S(-) enantiomer of the latter (WIN 55,212-3) and the CB1 receptor antagonist N piperidino-5-(4-chlorophenyl)- 1-(2,4-dichlorophenyl)-4-methyl-3-pyrazole carboxamide (SR 141716). The electrically evoked tritium overflow from guinea-pig hippocampal slices was reduced by WIN 55,212-2 (pIC30% 6.5) but not affected by WIN 55,212-3 up to 10 microM. The concentration-response curve of WIN 55,212-2 was shifted to the right by SR 141716 (0.032-microM) (apparent pA2 8.2), which by itself did not affect the evoked overflow. WIN 55,212-2 1 microM also inhibited the Ca(2+)-evoked tritium overflow in guinea-pig hippocampal slices and the electrically evoked overflow in guinea-pig cerebellar, cerebrocortical and hypothalamic slices as well as in human hippocampal slices but not in rat and mouse hippocampal slices. SR 141716 (0.32 microM) markedly attenuated the WIN 55,212-2-induced inhibition in guinea-pig and human brain slices. SR 141716 0.32 microM by itself increased the electrically evoked tritium overflow in guinea-pig hippocampal slices but failed to do so in slices from the other brain regions of the guinea-pig and in human hippocampal slices but failed to do so in slices from the other brain regions of the guinea-pig and in human hippocampal slices. The cAMP accumulation stimulated by forskolin was reduced by CP-55,940 and WIN 55,212 2. The concentration-response curve of CP 55,940 was shifted to the right by SR 141716 (0.1 microM; apparent pA2 8.3), which by itself did not affect cAMP accumulation. In conclusion, cannabinoid receptors of the CB1 subtype occur in the human hippocampus, where they may contribute to the psychotropic effects of cannabis, and in the guinea-pig hippocampus, cerebellum, cerebral cortex and hypothalamus. The CB1 receptor in the guinea-pig hippocampus is located presynaptically, is activated by endogenous cannabinoids and may be negatively coupled to adenylyl cyclase. PMID- 9402038 TI - Caffeine induces central cholinergic analgesia. AB - The antinociceptive effect of caffeine was examined by using the hot-plate, abdominal constriction tests in mice and the tail flick and paw-pressure tests in rats. Caffeine (1-5 mg kg-1 s.c. in mice; 2.5-5 mg kg-1 i.p. in rats) produced significant antinociception in both species which was prevented by atropine (5 mg kg-1 i.p.), pirenzepine (0.1 microgram per mouse i.c.v.), hemicholinium-3 (1 microgram/ mouse i.c.v.) and N6-cyclopentyladenosine (5 micrograms/mouse i.c.v.), but not by naloxone (1 mg kg-1 i.p.), CGP 35348 (100 mg kg-1 i.p.), alpha-methyl p-tyrosine (100 mg kg-1 i.p.) and reserpine (2 mg kg-1 i.p.). Intracerebroventricular injection of caffeine in mice at doses (2.5-5 micrograms per mouse) which were largely ineffective by parenteral routes, induces an antinociception whose intensity equalled that obtainable s.c. or i.p. In the antinociceptive dose-range, caffeine did not produce any behavioural impairment as revealed by the rotarod and Irwing tests. On the basis of the above data, it can be postulated that caffeine exerts an antinociceptive effect mediated by central amplification of cholinergic transmission. PMID- 9402039 TI - Antinociceptive action of DBO 17 and DBO 11 in mice: two 3,8 diazabicyclo (3.2.1.) octane derivates with selective mu opioid receptor affinity. AB - Two 3,8 diazabicyclo (3.2.1.) octane derivates, namely DBO 17 and DBO 11, were studied for the opioid-like activity. In the rat brain membrane preparation binding studies, DBO 17 and DBO 11 showed a high affinity and selectivity for the mu opioid receptor (Ki's: 5.1 and 25 nM, respectively). DBO 17 and DBO 11 inhibited the nociceptive response in the hot-plate test of mice with ED50 values of 0.16 mg/kg and 0.44 mg/kg, respectively. The antinociceptive action of both DBO 17 and DBO 11 was blocked by naloxone. Tolerance to the antinociceptive action of DBO 17 and DBO 11 was present after 13 and 7 days of repeated treatment, respectively. Both DBO 17 and DBO 11 were ineffective in morphine tolerant mice and vice versa. Chronic treatments (three times daily for seven consecutive days) of DBO 17 and DBO 11 induced a naloxone-precipitated withdrawal syndrome in DBO 17 treated mice similar to that in morphine treated mice, whereas in DBO 11 treated mice abstinence signs were virtually absent. These results indicate an interesting pharmacological profile that suggests these compounds as possible new candidates for the clinical treatment of pain. PMID- 9402040 TI - Differential regulation of corticotropin-releasing factor and vasopressin in discrete brain regions after morphine administration: correlations with hypothalamic noradrenergic activity and pituitary-adrenal response. AB - The changes in the content of corticotropin-releasing factor (CRF) and arginine vasopressin (AVP) in discrete brain nuclei during chronic opioids administration have not been well established. We evaluated the effects of acute and chronic morphine administration on the content of CRF and AVP in different hypothalamic and extrahypothalamic (bed nucleus of the stria terminalis, BNST) nuclei in rats. Concomitantly, changes in hypothalamic noradrenaline (NA) turnover [estimated by the 3-methoxy-4-hydroxyphenylethyleneglycol MHPG/NA ratio] and in plasma corticosterone release (as a marker of the activity of the hypothalamus-pituitary adrenal axis) were determined. Male rats were implanted with placebo (naive) or morphine (tolerant) pellets for 7 days. On day 8, groups of rats received an acute injection of either saline i.p. or morphine (30 mg/kg i.p.) and were sacrificed 30 min later. Acute morphine injection to naive rats increased both the release of corticosterone and the hypothalamic NA turnover. CRF and AVP showed no modifications in the paraventricular nucleus (PVN) or in the median eminence (ME). CRF content decreased in the ventromedian nucleus (VMN) and increased in the BNST, but did not change in the arcuate nucleus (AN). AVP was elevated in the supraoptic nucleus (SON) but not changed in the suprachiasmatic nucleus (SCN). In chronic morphine-treated rats, there was a pronounced decrease in the NA turnover and in the release of corticosterone, which indicates that tolerance develops to the acute effects of morphine. Correspondingly, CRF and AVP were enhanced in the PVN and decreased in the ME, when compared with naive rats injected with morphine. CRF content was decreased in the AN and in the BNST, but increased in the VMN. The AVP content was decreased in the SON, and no modifications were seen in the SCN. The present study shows that, in addition to the modifications in corticosterone secretion and in hypothalamic NA turnover, chronic morphine administration produces a complex response in the CRF and AVP systems. These modifications might contribute to the behavioral, emotional and neuroendocrine alterations produced during opioid tolerance. PMID- 9402042 TI - Methylene blue induces ongoing activity in rat cutaneous primary afferents and depolarization of DRG neurons via a photosensitive mechanism. AB - The dye methylene blue is known as a blocker of guanylyl cyclase and it has been widely used to deplete cells of internal cyclic GMP. The data presented demonstrate an activation of adult rat sensory neurons by methylene blue via a photosensitive mechanism. In single fiber recordings from primary afferents of the rat skin in vitro, methylene blue, applied to the receptive field, induced discharge activity: 2/2 A beta-, 2/4 A delta- and 5/7 C-fibers showed significantly enhanced firing upon 10 microM methylene blue in the presence of light, whereas the dye was ineffective when illumination was prevented. In whole cell current clamp experiments with dissociated dorsal root ganglion neurons, 100 microM methylene blue was ineffective in the dark but evoked a membrane depolarization of 15.3 +/- 3.5 mV (n = 5) accompanied by discharge activity upon illumination. In whole cell voltage clamp experiments, methylene blue (100 microM) caused a significant slowing of the inactivation of voltage-dependent sodium currents. In addition, an inhibition of fast and slow outward currents was observed with prolonged exposure. The impeded sodium inactivation together with the blockade of potassium currents may contribute to the depolarization and discharge activity observed in primary afferents in vitro as well as in dissociated sensory neurons in culture. We therefore suggest that methylene blue studies with excitable cells or tissues need to be interpreted with caution. PMID- 9402041 TI - Levetiracetam (ucb LO59) affects in vitro models of epilepsy in CA3 pyramidal neurons without altering normal synaptic transmission. AB - Previous behavioural and electrophysiological studies have indicated that levetiracetam (ucb LO59) acts as an anticonvulsant drug in vivo. The purpose of the present study was to investigate the effects of levetiracetam on normal synaptic transmission and epileptiform activity in vitro. Intracellular recordings were obtained from the CA3 subfield of the rat hippocampal slice preparation. Levetiracetam in a concentration of 10 microM did not influence basic cell properties or normal synaptic transmission evoked by subthreshold and suprathreshold stimuli to the commissural pathway. However, it strongly inhibited the development of epileptiform bursting by the gamma-aminobutyric acid (GABA)A receptor antagonist bicuculline (1-30 microM). Levetiracetam also decreased the size of bursts previously established by bicuculline. In experiments in which the glutamate-receptor agonist N-Methyl-D-Aspartate (NMDA) was used to generate spontaneous bursting, levetiracetam had no effect on the size of the bursts but decreased bursting frequency. The difference in effects of levetiracetam on bicuculline- and NMDA-induced bursting appeared to be dependent on the convulsant used, since in the presence of 10 microM bicuculline, levetiracetam decreased the size of NMDA-bursts to the same extent as the size of synaptically evoked bicuculline-bursts but had little effect on bursting frequency. The results show that under our experimental conditions, levetiracetam did not alter the components of normal synaptic transmission. However, levetiracetam at the concentrations studied inhibited epileptiform bursting induced by bicuculline and NMDA in vitro in a manner consistent with the profile of an antiepileptogenic drug. PMID- 9402043 TI - Presynaptic snake beta-neurotoxins produce tetanic fade and endplate potential run-down during neuromuscular blockade in mouse diaphragm. AB - The present study investigated the ability of a number of presynaptic snake neurotoxins (snake beta-neurotoxins) to produce nerve-evoked train-of-four fade, tetanic fade and endplate potential run-down during the development of neuromuscular blockade in the isolated mouse phrenic-hemidiaphragm nerve-muscle preparation. All the snake beta-neurotoxins tested, with the exception of notexin, produced train-of-four and tetanic fade of nerve-evoked isometric muscle contractions. Train-of-four fade was not present during the initial depressant or facilitatory phases of muscle tension produced by the snake beta-neurotoxins but developed progressively during the final depressant phase that precedes complete neuromuscular blockade. The 'non-neurotoxic' bovine pancreatic phospholipase A2 and the 'low-toxicity' phospholipase A2 from Naja naja atra venom failed to elicit train-of-four fade, indicating that the phospholipase activity of the snake beta-neurotoxins is not responsible for the development of fade. Intracellular recording of endplate potentials (EPPs) elicited by nerve-evoked trains of stimuli showed a progressive run-down in EPP amplitude during the train following incubation with all snake beta-neurotoxins except notexin. Again this run-down in EPP amplitude was confined to the final depressant phase of snake beta-neurotoxin action. However when EPP amplitude fell to near uniquantal levels (< 3 mV) the extent of toxin induced-fade was reduced. Unlike postjunctional snake alpha-neurotoxins, prejunctional snake beta-neurotoxins interfere with acetylcholine release at the neuromuscular junction during the development of neuromuscular blockade. This study provides further support for the hypothesis that fade in twitch and tetanic muscle tension is due to an underlying rundown in EPP amplitude resulting from a prejunctional alteration of transmitter release rather than a use-dependent block of postjunctional nicotinic receptors. PMID- 9402044 TI - The Y1 antagonist BIBP 3226 inhibits potentiation of methoxamine-induced vasoconstriction by neuropeptide Y. AB - We investigated the interaction of neuropeptide Y (NPY) with the alpha 1 adrenoceptor agonist, methoxamine, in control of mean arterial pressure, renovascular resistance and mesenteric vascular resistance in anaesthetized rats. Infusion of 3.0 but not 0.3 microgram/kg/min NPY enhanced the elevations of all three haemodynamic parameters caused by bolus injections of methoxamine (10-100 micrograms/kg). These enhancements largely involved a prolongation of the methoxamine effects. While infusion of the Y1 NPY receptor-selective antagonist, BIBP 3226 (10 micrograms/kg/min), alone did not alter methoxamine-induced vasoconstriction, it inhibited the potentiation by NPY. We conclude that NPY can potentiate methoxamine-induced vasoconstriction in vivo. This is mediated predominantly, if not exclusively, via the Y1 receptor. Endogenously released NPY does not appear to reach sufficient concentrations to cause tonic systemic vasoconstriction or potentiation thereof in the anaesthetized rat. PMID- 9402045 TI - Characterization by antagonists of P2-receptors mediating endothelium-dependent relaxation in the rat aorta. AB - The receptors through which 2-methylthio ATP (MeSATP), adenosine 5'-O-(2 thiodiphosphate) (ADP beta S), UTP and ATP elicit endothelium-dependent relaxation of noradrenaline-precontracted rings of the rat aorta were characterized by means of a series of antagonists. The acetylcholine-induced relaxation and the degradation of MeSATP, UTP and ATP were also studied. The potency of the nucleotides at producing relaxation decreased in the order MeSATP (EC50 0.24 microM) > ADP beta S (0.43 microM) > UTP (1.09 microM) > ATP (3.53 microM). MeSATP, ADP beta S and UTP did not cause relaxation when the endothelium had been destroyed; high concentrations of ATP still caused some relaxation. The relaxation by MeSATP, ADP beta S and UTP became very small after treatment of the rings with NG-nitro-L-arginine methyl ester; the relaxation by ATP was less affected. Pre-exposure to MeSATP (100 microM) abolished or almost abolished the relaxation normally elicited by MeSATP and ADP beta S, did not change that elicited by UTP and slightly enhanced the relaxation elicited by ATP. Of nine compounds examined as antagonists, six attenuated selectively the effect of some or all of the nucleotides (as compared to acetylcholine): suramin, reactive blue 2, pyridoxalphosphate-6-azophenyl-2',5'-disulphonate (iso-PPADS), pyridoxalphosphate-6-azophenyl-2',4'-disulphonate (PPADS), reactive red 2 and 5,5'-(1,1'-biphenyl-4,4'-diylbisazo)-bis-7-amino -6-hydroxy-naphthalene-1,4 disulphonate (NH05). Decreases of maximal relaxations and slopes different from unity in Schild plots often indicated non-competitive kinetics of the antagonism. For each of the six 'selective' antagonist, the apparent Kd values against MeSATP and against ADP beta S were similar: none of the six differentiated between MeSATP and ADP beta S. Also, for each of four 'selective' antagonists, the apparent Kd values against UTP and against ATP were similar: none of the four differentiated between these two nucleotides (two antagonists did not act against UTP and ATP in the 'selective' concentration range). On the other hand, for five of the six 'selective' antagonists (the exception being NH05), the apparent Kd values against MeSATP and ADP beta S were considerably lower than those against UTP and ATP. At the highest concentrations tested against agonist-evoked relaxations, the antagonists did not alter the removal from the incubation medium, by pieces of rat aorta, of MeSATP, UTP and ATP. It is concluded that nucleotides cause endothelium-dependent relaxation of the rat aorta through two sites: a P2Y-receptor and a P2U-receptor. The receptors may be pharmacologically similar to a bovine endothelial P2Y (P2Y1) and a cloned rat P2U (P2Y2) receptor, respectively. ATP acts mainly through the P2U-receptor. Suramin, reactive blue 2, iso-PPADS, PPADS and reactive red 2 are more potent at the P2Y- than the P2U receptor. NH05 does not discriminate between the two receptors but is the most potent P2U antagonist so far described. PMID- 9402046 TI - Vasoconstrictor and vasodilator effects of guanine nucleotides in the rat aorta. AB - Although GTP, like ATP and UTP, is stored in platelet dense granules, little is known about its vascular effects. The present study was carried out in order to characterize the effects of GTP and related compounds in the rat aorta. Contractions were examined in aortic rings at resting tension. In rings with intact endothelium, GTP, GDP, guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S) and guanosine 5'-O-(2-thiodiphosphate) (GDP beta S) caused small contractions. In endothelium-denuded rings, the contractions were unchanged or increased and persisted after desensitization of P2X-receptors by alpha,beta-methylene ATP. Relaxations were examined in aortic rings precontracted with noradrenaline. In rings with intact endothelium, GTP (EC50 131 microM), GDP (no maximal effect obtained), GTP gamma S (EC50 6.8 microM) and guanosine (EC50 822 microM) caused prominent relaxation, whereas GDP beta S caused further contraction. In endothelium-denuded rings, the relaxant effect of GTP was greatly reduced, that of GDP and guanosine was unchanged, and that of GTP gamma S was abolished. Relaxations by GTP and GTP gamma S in endothelium-intact rings were studied in more detail. The relaxation by GTP was slightly and the relaxation by GTP gamma S greatly reduced after treatment with NG-nitro-L-arginine methyl ester. Pre exposure to a high concentration of the P2Y-receptor agonist 2-methylthio ATP (MeSATP) did not attenuate the effects of GTP and GTP gamma S. Four compounds previously identified as antagonists at the P2Y- and P2U-receptors of rat aortic endothelium--suramin, reactive blue 2, pyridoxalphosphate-6-azophenyl-2',5' disulphonate (iso-PPADS) and 5,5'-(1,1'-biphenyl-4,4'-diylbisazo)-bis-7-amino-6- hydroxynaphthalene-1,4-disulphonate (NH05)--were tested against GTP and GTP gamma S. Suramin, reactive blue 2 and iso-PPADS were much less potent against GTP and GTP gamma S than previously found against (the P2Y effect of) MeSATP. Suramin, iso-PPADS and NH05 were about as potent against GTP and GTP gamma S as previously found against (the P2U effect of) UTP and in particular ATP. It is concluded that guanine nucleotides can cause both contraction and relaxation of the rat aorta. The high concentrations of GTP and GDP required, and in the case of contraction the small size of the response, make a physiological role of the vascular effects of these nucleotides unlikely. GTP and GTP gamma S elicit endothelium-dependent relaxation through P2U-receptors. GTP in addition relaxes the aorta through smooth muscle receptors, possibly by way of its degradation product guanosine. The stable analog GTP gamma S is a relatively potent and selective agonist for the endothelial P2U-receptor. PMID- 9402047 TI - Kinin B1 and B2 receptors in pig vessels: characterization of two monoreceptor systems. AB - The coronary artery and renal vein of the adult pig are sensitive and reliable monoreceptor systems for studying kinin receptors. The pig coronary artery with intact endothelium is highly sensitive to bradykinin (BK, pEC50 8.6), while being insensitive to the B1 receptor agonist, LysdesArg9BK. The tissue responds to BK with concentration-dependent relaxation, which is prevented by B2 receptor antagonists, particularly DArg[Hyp3, Thi5, DTic7, Oic8]BK (HOE 140, pKB 9.3), (E) 3-(6-acetoamido-3-pyridyl)-N-(N-{2, 4-dichloro-3-[(2-methyl-8-quinolinyl)oxy methyl]phenyl}-N- methylaminocarbonyl-methyl)acrylamide (FR 173657), a new non peptide compound (pKB 9.3), while B1 receptor antagonists (e.g. Lys[Leu8]desArg9BK) are inactive. The order of potency of kinin-related peptides in this vessel is: LysBK > or = BK > [Hyp3]BK > [Aib7]BK, a sequence typical of a B2 receptor system. Antagonists such as HOE 140 and FR 173657, at high concentrations reduce the maximum effect of BK and thus behave as noncompetitive antagonists. The kinin B1 receptor was studied in the pig renal vein without endothelium and incubated for several hours in order to allow for the de novo formation of this functional site. After 7-8 h in vitro incubation, the vessel shows high sensitivity to LysdesArg9BK (pEC50 8.3) and is insensitive to BK. The pig renal vein responds to B1 receptor agonists with concentration-dependent contraction which maintains a stable plateau and is prevented by selective B1 receptor antagonists such as Lys[Leu8]desArg9BK (pKB 6.7). The most active antagonist has been found to be desArg9HOE 140 (pA2 7.6) which acts as competitive antagonist in this preparation. Some B2 antagonists (e.g. HOE 140) show weak (pKB 6.1) anti-B1 receptor activity while the non-peptide compound FR 173657 is inactive on the B1 receptor and therefore acts as a potent and selective kinin B2 receptor antagonist in the pig. The data obtained in this study allow us to compare the porcine B2 and B1 receptors with those of other species including man, and underline some interesting features that are unique to the porcine functional sites. PMID- 9402048 TI - Characterization of postjunctional muscarinic receptors mediating contraction in rat anococcygeus muscle. AB - The present study was designed to characterize the postjunctional muscarinic receptors mediating contraction in rat anococcygeus muscle by means of a series of muscarinic agonists and subtype-preferring key muscarinic antagonists. Cumulative addition of muscarinic agonists elicited concentration-dependent contractions with the following rank order of potency (pD2 values): (+)-muscarine (6.36) > or = oxotremorine M (6.21) > or = arecaidine propargyl ester (APE) (6.18) > carbachol (5.68) = (+/-)-methacholine (5.65) > 4-(4-chlorophenyl carbamoyloxy)-2-butynyltrimethylammonium chloride (4-Cl-McN-A-343) (4.28) > 4-(3 chlorophenylcarbamoyloxy)-2-butynyltrimethylammonium chloride (McN-A-343) (3.89). (+)-Muscarine, oxotremorine M, carbachol and (+/-)-methacholine behaved as full agonists, whereas APE, 4-Cl-McN-A-343 and McN-A-343 displayed partial agonism. The contractile responses of the rat anococcygeus muscle to (+/-)-methacholine were competitively antagonized by pirenzepine (pA2 = 6.92), 11-[[4-[4 (diethylamino)butyl]-1-piperidinyl]acetyl] 5,11-dihydro-6H-pyrido(2,3-b) (1,4) benzodiazepine-6-one (AQ-RA 741; pA2 = 6.75), himbacine (pA2 = 7.11), (+/-)-p fluoro-hexahydro-sila-difenidol (p-F-HHSiD; pA2 = 7.68) and the (R)- and (S) enantiomers of hexahydro-difenidol [(R)-HHD: pA2 = 8.52; (S)-HHD: pA2 = 6.06]. A comparison of the pA2 values derived from studies of contraction in rat anococcygeus muscle with literature binding (pKi values) and functional affinities (pA2 values) obtained at native M1-M4 receptors strongly suggests that the postjunctional muscarinic receptors mediating contraction in rat anococcygeus muscle are of the M3 subtype. PMID- 9402049 TI - MEN 11420, a potent and selective tachykinin NK2 receptor antagonist in the guinea-pig and human colon. AB - We have characterized the action of the novel, water-soluble, tachykinin NK2 receptor antagonist MEN 11420 ([Asn(2-AcNH-beta-D-Glc)-Asp-Trp-Phe-Dap-Leu] c(2 beta-5 beta)) on the circular muscle of the guinea-pig and human colon in vitro and on the guinea-pig colon in vivo. In organ bath experiments on guinea-pig colon MEN 11420 produced a concentration-dependent rightward shift of the concentration-response curve to the NK2 receptor selective agonist, [beta Ala8]neurokinin A (NKA) (4-10) with a pKB value of 8.1. Up to 1 microM MEN 11420 had no effect on the concentration-response curve to methacholine, to the NK1 receptor selective agonist, [Sar9]substance P (SP) sulfone, to the NK3 receptor selective agonist, senktide, or on the response to exogenous SP. The response to exogenous NKA was inhibited, although the shift of the concentration-response curve to NKA produced by MEN 11420 at 1 microM (dose ratio 5.3) was much smaller than that produced against [beta Ala8]NKA (4-10) (dose ratio 102), presumably because NKA also stimulates NK1 receptors at relatively low concentrations. In sucrose gap, MEN 11420 concentration-dependently inhibited both depolarization (IC50 0.34 microM) and contraction (IC50 = 0.32 microM) produced by [beta Ala8]NKA (4-10) (0.3 microM for 10 s) in the guinea-pig colon without affecting the corresponding responses produced by [Sar9]SP sulfone. When similar experiments were performed in the circular muscle of the human colon MEN 11420 concentration-dependently inhibited both depolarization and contraction induced by [beta Ala8]NKA(4-10) with IC50s of 99 and 75 nM, respectively. MEN 11420 (1 microM) had no effect on the nonadrenergic noncholinergic (NANC) depolarization and contraction produced by a short period of electrical field stimulation (EFS, 10 Hz for 1 s) in the guinea-pig colon and selectively inhibited the sustained component of depolarization produced during a prolonged period of EFS (3 Hz for 3 min), without affecting the concomitant depolarization. Nifedipine (1 microM) eliminated the NANC contraction to a short period of EFS and the phasic contraction in response to a prolonged period of EFS. MEN 11420 (1 microM) abolished the nifedipine-resistant NANC contraction produced by prolonged period of electrical field stimulation (EFS, 3 Hz for 3 min). All electrical and mechanical NANC responses to EFS which were resistant to MEN 11420, either in the absence or presence of nifedipine, were abolished by the subsequent application of the NK1 receptor antagonist, SR 140333 (1 microM). Up to 3 microM, MEN 11420 had no significant effect on the cholinergic excitatory junction potential or the NANC inhibitory junction potential evoked by single pulse EFS, nor did it affect membrane conductance. In urethane-anaesthetized guinea-pigs MEN 11420 (10-100 nmol/kg i.v.) produced a dose-dependent and long lasting (> 3 h) inhibition of the contractile response (15 +/- 2 mmHg) of the proximal colon induced by [beta Ala8]NKA (4-10) (3 nmol/kg i.v.). MEN 11420 (300 nmol/kg i.v.) did not affect the contraction produced by [Sar9]SP sulfone. MEN 11420 (300 nmol/kg) produced a limited (Emax about 40% inhibition) and transient (recovery within 60 min) inhibition of the atropine- and hexamethonium-sensitive phasic contractions of the proximal colon induced by threshold distension of a colonic balloon. On the other hand, MEN 11420 (10-300 nmol/kg i.v.) produced a dose-dependent complete and prolonged (> 2 h from administration) inhibition of the atropine-resistant and hexamethonium-sensitive phasic contraction induced by suprathreshold distension of the colonic balloon. We conclude that MEN 11420 is a potent and selective tachykinin NK2 receptor antagonist devoid of significant inhibitory activity toward excitatory transmission mediated via tachykinin NK1 or muscarinic receptors. The present findings indicate that SP and NKA are likely involved in the preferential activation of NK1 and NK2 receptors during tachykininergi PMID- 9402051 TI - Mammalian glial cells in culture synthesize acetylcholine. AB - In the present study we demonstrate that acetylcholine is synthesized by cultured mammalian glial cells identified by cell-type specific markers. Primary cultures of rat brain astrocytes or microglia contained 2.0 and 1.6 pmol acetylcholine/10(6) cells on average respectively. Astrocyte cultures established from neonatal mouse brain contained even more acetylcholine (about 80 pmol acetylcholine/10(6) cells). Primary cultures of rat brain astrocytes showed choline acetyltransferase (ChAT) enzyme activity of 3 nmol/mg protein/h; ChAT activity was blocked by 10 microM bromoacetylcholine. In conclusion, these data demonstrate the synthesis of the "neurotransmitter" acetylcholine in cultured glial cells, a finding which opens a new view upon the role of acetylcholine in mammalian brain. PMID- 9402050 TI - NK1- and NK3-receptor mediated inhibition of 5-hydroxytryptamine release from the vascularly perfused small intestine of the guinea-pig. AB - The effects of tachykinins on the spontaneous release of 5-hydroxytryptamine (5 HT) from the enterochromaffin cells into the portal circulation was investigated in vitro using the vascularly perfused isolated guinea-pig small intestine. 5-HT was determined by HPLC with electrochemical detection. Test substances were applied intraarterially. Substance P (SP) caused a concentration-dependent decrease in 5-HT outflow with an EC50 of 50 pmol/l. Similarly, the selective NK1 receptor agonist SP methyl ester (1 nmol/l) significantly inhibited 5-HT outflow (to 51 +/- 3%). When tetrodotoxin (1 mumol/l) was added to the arterial perfusion medium, the inhibition by SP of 5-HT outflow was not affected. The selective NK1 receptor antagonist CP 99994 [(+)-(2S,3S)-3-(2-methoxybenzylamino)-2 phenylpiperidine] (0.1 mumol/l) prevented the inhibitory effect of SP (0.1 mumol/l). Neither GR 94800 (PhCO-Ala-Ala-DTrp-Phe-DPro-Pro-NleNH2) (0.1 mumol/l) nor SR 142801 [(S)-(N)-(1-(3-(1-benzoyl-3-(3,4-dichlorophenyl) piperidin-3 yl)propyl)-4-phenylpiperidin-4-yl)-N- methylacetamide] (10 nmol/l), which are selective NK2 and NK3 receptor antagonists, changed the SP-mediated inhibition. The selektive NK3 receptor agonist senktide (10 nmol/l) also decreased the 5-HT outflow (to 57 +/- 5%). This inhibition was prevented by SR 142801 (10 nmol/l) and by tetrodotoxin. CP 99994 (0.1 mumol/l) significantly antagonized the senktide-mediated inhibition of 5-HT outflow. The outflow of 5-HT was unaffected when CP 99994, GR 94800 or SR 142801 alone were added to the perfusion medium. It is concluded that the release of 5-HT from enterochromaffin cells is directly inhibited by NK1 receptors, and indirectly by neuronal NK3 receptors whose stimulation leads to the release of SP. PMID- 9402065 TI - The crystal structure of a type I cohesin domain at 1.7 A resolution. AB - The quaternary organization of the cellulosome, a multi-enzymatic extracellular complex produced by cellulolytic bacteria, depends on specific interactions between dockerin domains, double EF-hand subunits carried by the catalytic components, and cohesin domains, individual receptor subunits linearly arranged within a non-catalytic scaffolding polypeptide. Cohesin-dockerin complexes with distinct specificities are also thought to mediate the attachment of cellulosomes to the cell membrane. We report here the crystal structure of a single cohesin domain from the scaffolding protein of Clostridium thermocellum. The cohesin domain folds into a nine-stranded beta-sandwich with an overall "jelly roll" topology, similar to that observed in bacterial cellulose-binding domains. Surface-exposed patches of conserved residues promote extensive intermolecular contacts in the crystal, and suggest a possible binding target for the EF-hand pair of the cognate dockerin domain. Comparative studies of cohesin domains indicate that, in spite of low sequence similarities and different functional roles, all cohesin domains share a common nine-stranded beta-barrel fold stabilized by a conserved hydrophobic core. The formation of stable cohesin dockerin complexes requires the presence of Ca2+. However, the structure of the cohesin domain reported here reveals no obvious Ca2+-binding site, and previous experiments have failed to detect high affinity binding of Ca2+ to the unliganded dockerin domain of endoglucanase CelD. Based on structural and biochemical evidence, we propose a model of the cohesin-dockerin complex in which the dockerin domain requires complexation with its cohesin partner for protein stability and high-affinity Ca2+ binding. PMID- 9402066 TI - The 1.8 A crystal structure of the dimeric peroxisomal 3-ketoacyl-CoA thiolase of Saccharomyces cerevisiae: implications for substrate binding and reaction mechanism. AB - The dimeric, peroxisomal 3-ketoacyl-CoA thiolase catalyses the conversion of 3 ketoacyl-CoA into acyl-CoA, which is shorter by two carbon atoms. This reaction is the last step of the beta-oxidation pathway. The crystal structure of unliganded peroxisomal thiolase of the yeast Saccharomyces cerevisiae has been refined at 1.8 A resolution. An unusual feature of this structure is the presence of two helices, completely buried in the dimer and sandwiched between two beta sheets. The analysis of the structure shows that the sequences of these helices are not hydrophobic, but generate two amphipathic helices. The helix in the N terminal domain exposes the polar side-chains to a cavity at the dimer interface, filled with structured water molecules. The central helix in the C-terminal domain exposes its polar residues to an interior polar pocket. The refined structure has also been used to predict the mode of binding of the substrate molecule acetoacetyl-CoA, as well as the reaction mechanism. From previous studies it is known that Cys125, His375 and Cys403 are important catalytic residues. In the proposed model the acetoacetyl group fits near the two catalytic cysteine residues, such that the oxygen atoms point towards the protein interior. The distance between SG(Cys125) and C3(acetoacetyl-CoA) is 3.7 A. The O2 atom of the docked acetoacetyl group makes a hydrogen bond to N(Gly405), which would favour the formation of the covalent bond between SG(Cys125) and C3(acetoacetyl CoA) of the intermediate complex of the two-step reaction. The CoA moiety is proposed to bind in a groove on the surface of the protein molecule. Most of the interactions of the CoA molecule are with atoms of the loop domain. The three phosphate groups of the CoA moiety are predicted to interact with side-chains of lysine and arginine residues, which are conserved in the dimeric thiolases. PMID- 9402068 TI - Elevated serum macrophage inhibitory factor-related protein (MRP) 8/14 levels in advanced HIV infection and during disease exacerbation. AB - To assess the value of MRP 8, MRP 14, and MRP 8/14 serum concentrations as markers of disease progression in HIV infection and as markers of intercurrent infections. DESIGN: We measured MRP 8, MRP 14, and MRP 8/14 serum concentrations in 184 HIV-infected patients in various stages of disease with or without disease exacerbation and in 50 healthy control subjects. In clinically stable HIV infection correlations of MRP levels with stage of HIV disease, CD4 counts, p24 antigen, and beta-2 microglobulin levels were studied. In patients with intercurrent illnesses, correlations of MRP levels with type of disease exacerbation and with CRP were calculated and compared with those found in stable HIV infection. RESULTS: MRP 8/14 levels were significantly elevated and MRP 8 levels slightly decreased in stable HIV infection compared with HIV-negative controls. The CD4 cell count and MRP 8/14 levels correlated significantly in patients with AIDS. Despite higher values of MRP 8/14 during advanced disease, these were not significant predictors of progression to death. In patients with acute infections, MRP 8/14 levels were significantly elevated, compared with patients with illnesses of noninfectious origin. Levels of MRP 8/14 associated with acute infections were significantly higher in patients with AIDS than in patients during earlier stages of HIV infection. CONCLUSIONS: Both stable HIV infection and advanced immunedeficiency are associated with an elevation of the MRP 8/14 complex and probably with a decline of MRP 8 serum levels. MRP 8/14 is preserved as a marker of acute infection in immunecompromised patients. PMID- 9402067 TI - Antioxidants and dipyridamole inhibit HIV-1 gp120-induced free radical-based oxidative damage to human monocytoid cells. AB - Reactive oxygen species (ROS) may play an important role in HIV-1 pathogenesis and HIV-1 gp120-induced neurotoxicity. Our studies determined the extent to which gp120 increased ROS production in human monocytic U937 cells and the effectiveness of various agents, including dipyridamole (DPR), in blocking these responses. The thiobarbituric acid-reactive substances (TBARS) assay was used as a measure of recombinant gp120 (HIV-1[3B])-induced oxidative damage to U937 cells. As a control, TBARS production was measured using a hypoxanthine/xanthine superoxide generating system. There was gp120-induced oxidative damage in U937 cells with a concentration that produces 50% of maximal effect (apparent EC50 value) of 11 pM. Polyclonal antiserum to gp120 significantly (p < 0.05) inhibited gp120-induced oxidative damage. gp120-induced oxidative damage was significantly inhibited 81% (p < 0.01) by catalase/superoxide dismutase, 53% (p < 0.05) by (+/ )-alpha-tocopherol, 78% (p < 0.01) by desferrioxamine, and 82% (p < 0.01) by ethylene diamine tetraacetic acid (EDTA). These results indicate that gp120 is capable of promoting iron-based oxygen free radical damage to U937 cells. DPR potently (p < 0.05) inhibited both hypoxanthine/xanthine- and gp120-induced oxidative damage with concentrations that produce 50% inhibition (apparent IC50 values) of 1.3 microM for hypoxanthine/xanthine and 1.0 microM for gp120. Therapeutic intervention against ROS production may prevent HIV-1 neurotoxicity. PMID- 9402069 TI - CCR5del32 in perinatal HIV-1 infection. AB - CCR5, a chemokine receptor, serves as a coreceptor for macrophage-tropic HIV-1 (1 3). A 32-bp deletion within the gene encoding CCR5, CCR5del32, has been shown to prevent HIV-1 infection of T cells in the absence of a wild-type allele. This alteration is present in low frequency in Caucasian populations (4-6). To investigate the effect of CCR5del32 in perinatal HIV-1 transmission and disease progression, two cohorts of perinatally exposed infected and uninfected children were analyzed for the presence of the allele. Polymerase chain reaction (PCR) was used to identify CCR5del32 in prevalent and prospective cases among 144 African American children from New York City and 73 Caucasian children from Barcelona, Spain. HIV-1 transmission; clinical manifestations of disease, including encephalopathy, opportunistic infections, and death before 2 years of age; survival; Centers for Disease Control and Prevention (CDC) classification; and degree of immunosuppression were compared in children with and without CCR5del32. The allele frequency in HIV-1-infected African Americans (0.016) was lower than in Catalan children (0.041). No evidence for a dominant protective effect of CCR5del32 for HIV-1 transmission or disease progression was found in these cohorts. PMID- 9402070 TI - Risk of mother-to-infant transmission of HIV-1 is not reduced in CCR5/delta32ccr5 heterozygotes. AB - To determine if the 32-bp deletion of the chemokine receptor CCR5 (delta32ccr5) protects against mother-to-infant transmission of HIV-1, specimens from all uninfected and infected children who were perinatally exposed to HIV-1 and observed since 1988 and whose mothers did not take zidovudine were assessed for delta32ccr5. The CCR5 genotype was determined using polymerase chain reaction (PCR) for 122 subjects, of whom 73 were HIV-1 infected and 49 were perinatally exposed but uninfected; 70% and 71%, respectively, were Caucasian. Eleven of 73 (15%) infected children and 4 of 49 (8%) exposed uninfected children were CCR5/delta32ccr5 heterozygotes (p = 0.40). Among subjects who had at least one Caucasian parent or grandparent, 11 of 51 (22%) HIV-1-infected persons and 4 of 35 (11%) uninfected persons were heterozygotes. None were homozygous for the delta32ccr5 allele. The estimated relative risk for mother-to-infant HIV-1 transmission in heterozygotes was 2.0. Furthermore, the 95% confidence interval (0.6, 7.3) suggested that it is unlikely that the true relative risk was <0.6. Thus, the infant CCR5/delta32ccr5 heterozygous genotype was not associated with a diminished risk of perinatally acquired HIV-1 infection. PMID- 9402072 TI - Testosterone replacement treatment options for HIV-infected men. AB - Hypogonadism is well documented in HIV-infected men, particularly as they progress to AIDS and in those with symptoms of wasting. Testosterone deficiency can be diagnosed with simple laboratory tests, and various treatment options exist. The benefits of androgen replacement are well documented from a large body of literature and experience with hypogonadal men without HIV infection. Hypogonadal men who are given testosterone replacement have improved sexual thoughts and functioning, more energy, and improved mood. Generally, quality of life improves with such therapy. Testosterone replacement tends to maintain or improve lean body mass. The benefit, dose, and timing of testosterone replacement treatment for men with HIV infection, however, are less clear and require further study. Appropriate history and a high degree of clinical suspicion, coupled with relatively simple laboratory measurements, can confirm the diagnosis of hypogonadism in men with HIV. Various options for testosterone replacement, including injections of testosterone esters and the use of transcutaneous patches, are discussed, as are the uses of pharmacologic doses of testosterone, primarily for its potential anabolic effect. PMID- 9402071 TI - Safety and antiviral activity of combination therapy with zidovudine, zalcitabine, and two doses of interferon-alpha2a in patients with HIV. AIDS Clinical Trials Group Study 197. AB - We conducted a three-arm, randomized, phase II study to evaluate the combination of zidovudine (600 mg/day) and zalcitabine (2.25 mg/day) alone or with one of two interferon-alpha2a doses (1 mIU or 6 mIU daily). Primary study endpoints included toxicity and changes from baseline for plasma HIV-1 RNA, CD4 cells, and quantitative microculture at weeks 8 and 24. Sixty-three patients with HIV infection and <400 CD4 cells/mm3 were enrolled; four patients discontinued therapy within 2 weeks. Adverse event rates were 37%, 32%, and 60%, respectively, for the nucleoside, 1-mIU interferon, and 6-mIU interferon combination groups. Increasing doses of interferon resulted in significantly greater hematologic toxicity (p = 0.03) and peripheral neuropathy (p = 0.02). Plasma HIV-1 RNA reductions were noted across all treatment groups at week 8 (p < 0.001) but only for the nucleoside and 1-mIU interferon combination groups at week 24 (p < 0.001). Mean reductions in HIV-1 RNA at week 8 were 0.94, 1.29, and 1.40 log10, respectively, for the nucleoside, 1-mIU interferon, and 6-mIU interferon combination groups (p = 0.05); no differences were noted at week 24. No differences in CD4 cell counts were seen. The addition of interferon-alpha2a to zidovudine and zalcitabine resulted in transient enhanced decreases in viral load and increased toxicity. PMID- 9402073 TI - Risk behavior for HIV infection in participants in preventive HIV vaccine trials: a cautionary note. AB - We conducted a longitudinal study of participants in phase I and II HIV vaccine safety and immunogenicity trials to examine changes in sexual risk behavior that are associated with risk of HIV transmission. The participants were 48 HIV negative men and women enrolled in one of two placebo-controlled HIV vaccine trials conducted at San Francisco General Hospital. There was a significant increase in insertive unprotected anal intercourse (UAI) from 9% at baseline (trial entry), to 13% at the month 6 assessment, to 20% at the month 12 assessment (p = .02). The primary predictor of either insertive or receptive UAI during the vaccine trials was having engaged in this behavior prior to entry (p = .001). Higher-risk behavior was also seen among participants who were younger and had multiple sexual partners (each, p = .06) and who indicated that one of their reasons for participation in the vaccine trial was hope of protection from HIV infection (p = .07). These findings indicate that despite instructions otherwise, participants with a history of high-risk behavior or who express hope of protection from HIV infection by enrolling in vaccine trials may be candidates for more intensive risk-behavior counseling prior to and during their participation. PMID- 9402074 TI - HIV-1 infection in women is associated with severe nutritional deficiencies. AB - Nutritional deficiencies may contribute to immune dysregulation, and have been shown to be sensitive markers of HIV-1 disease progression. Only limited information exists, however, regarding the nutritional profile of HIV-1 seropositive drug abusers. Immune and nutritional measurements were obtained in a subsample of 125 subjects from a larger cohort of drug users being followed for HIV-1 infection and cofactors of disease progression. Nutritional deficiencies, particularly vitamins A, E, and zinc, were widespread with up to 86% of the drug users exhibiting at least one nutritional alteration. Although immune parameters (CD4 count, CD8 count, beta2-microglobulin) were similar in the HIV-1-infected men and women, women had significantly poorer overall nutritional status, as measured by plasma proteins, which are considered to be sensitive markers of malnutrition. A comparison of individuals with advanced disease (CD4 count <200/mm3) revealed significantly lower levels of plasma prealbumin (p < .01), selenium, (p < .05), and greater deficiency of vitamins A (p < .01) and E (p < .05) in women than in men. The greater severity of nutritional deficiencies noted in HIV-1-infected women may be an important determinant of disease progression and survival. PMID- 9402075 TI - Relation between soluble CD30 levels measured soon after HIV seroconversion and disease progression in men with hemophilia. AB - Soluble CD30 (sCD30) levels within 3 years of HIV seroconversion were studied in 85 hemophilic men infected with HIV. All men were coinfected with hepatitis C virus (HCV). Levels of sCD30 were elevated in these men when compared with controls. These elevated levels did not appear to be a result of treatment with intermediate-purity clotting factor concentrates and were unlikely to be due to HCV coinfection inasmuch as hemophilic patients infected with HCV alone showed only mildly elevated sCD30 levels when compared with those of hemophilic controls uninfected with HCV. Initial sCD30 levels were not significantly associated with progression to any endpoint, although a tendency was present for those with the highest initial levels to progress less rapidly than those with lower values. Despite elevated sCD30 levels in these men, we have not been able to confirm that high sCD30 levels are associated with more rapid HIV progression. PMID- 9402076 TI - Mother-to-child transmission of HIV-1. AB - After reviewing the evidence on the relation of vertical transmission of HIV to stage of infection in the mother, I developed a stochastic model of transmission in which the probability of transmission per week is proportional to the virus load in the mother. The virus load in different stages of the infection is measured by viral RNA levels or tissue culture infectious virus levels in plasma. The constant of proportionality is assumed to be different for transmission during pregnancy, during parturition, and during breast-feeding. Using data on transmission from mothers who are in the primary stage of infection, I estimated the constant of proportionality and calculated the probability of transmission during pregnancy as a function of the time pregnancy starts in relation to the stage of the infection. For breast-feeding, I calculated the conditional probability of transmission by breast-feeding for 20 weeks, dependent on the infant escaping infection during pregnancy and parturition. As might be expected, the probabilities of transmission are highest if the mother is in the primary stage of infection or in late stages of the disease and is quite low when the mother is in the asymptomatic stage of the infection. PMID- 9402077 TI - The association of herpes simplex virus type 2 (HSV-2), Haemophilus ducreyi, and syphilis with HIV infection in young men in northern Thailand. AB - To evaluate the association between sexually transmitted diseases that commonly may cause genital ulceration and prevalent and incident HIV infections, we conducted three case control studies in a cohort of 21-year-old male military conscripts in northern Thailand. The men were evaluated at baseline in 1991 and semiannually until their discharge 2 years later. Serologic evidence of infection with herpes simplex virus type 2 (HSV-2), Haemophilus ducreyi, and HIV were more frequent at baseline in 83 men with a history of genital ulcer than in 97 men without such a history. Seropositivity to H. ducreyi (odds ratio [OR] = 3.46), HSV-2 (OR = 3.83), and syphilis (OR = 1.53) were more common in HIV-positive than HIV-negative men. Men (N = 45) who seroconverted to HIV while in the military were more often seropositive for H. ducreyi and HSV-2 before HIV seroconversion and also were more likely to seroconvert to HSV-2 and H. ducreyi during the same interval as their HIV seroconversion compared with men who remained HIV-negative. These data suggest that HSV-2 and H. ducreyi may be both markers for high-risk sexual behavior and risk factors for HIV infection among young men in Thailand. PMID- 9402078 TI - Staphylococcus aureus nasal colonization in HIV-seropositive and HIV-seronegative drug users. AB - Nasal colonization plays an important role in the pathogenesis of Staphylococcus aureus infections. To identify characteristics associated with colonization, we studied a cross-section of a well-described cohort of HIV-seropositive and seronegative active and former drug users considered at risk for staphylococcal infections. Sixty percent of the 217 subjects were Hispanic, 36% were women, 25% actively used injection drugs, 23% actively used inhalational drugs, 23% received antibiotics, and 35% were HIV-seropositive. Forty-one percent of subjects had positive nasal cultures for S. aureus. The antibiotic susceptibility patterns were similar to the local hospital's outpatient isolates and no dominant strain was identified by arbitrarily primed polymerase chain reaction (AB-PCR). Variables significantly and independently associated with colonization included antibiotic use (odds ratio [OR] = 0.37; confidence interval [CI] = 0.18-0.77), active inhalational drug use within the HIV-seropositive population (OR = 2.36; CI = 1.10-5.10) and female gender (OR = 1.97; CI = 1.09-3.57). Characteristics not independently associated included injection drug use, HIV status, and CD4 count. The association with active inhalational drug use, a novel finding, may reflect alterations in the integrity of the nasal mucosa. The lack of association between HIV infection and S. aureus colonization, which is contrary to most previous studies, could be explained by our rigorous control for confounding variables or by a limited statistical power due to the sample sizes. PMID- 9402079 TI - High cytomegalovirus antigenemia levels and cytomegalovirus syndrome in patients with AIDS. PMID- 9402080 TI - Presence of HTLV-I and HTLV-II infection in Honduras. PMID- 9402081 TI - Various types of injection equipment and risk of HIV infection. PMID- 9402082 TI - Early effect of aldosterone on the rate of synthesis of the epithelial sodium channel alpha subunit in A6 renal cells. AB - Transepithelial Na+ reabsorption across tight epithelia is regulated by aldosterone. The amiloride-sensitive epithelial sodium channel (ENaC) is a major target for the natriferic action of aldosterone. In this study, the effect of aldosterone on ENaC mRNA abundance and the rate of protein synthesis for each of the three ENaC subunits (alpha, beta and gamma) in the A6 kidney cell line were examined. In cells grown on plastic, aldosterone induced a large and rapid increase in epithelial sodium channel (ENaC) beta and gamma subunit mRNA abundance, but this effect is not translated into the synthesis of the corresponding proteins. In cells grown on a porous substrate, amiloride-sensitive electrogenic sodium transport was expressed and was upregulated by aldosterone (300 nM) as early as 1 h after the addition of the hormone. The alpha, beta, and gamma mRNA abundance was not changed by aldosterone during the first 3 h of stimulation, whereas a fourfold increase over control was observed after 24 h. The rate of synthesis of alpha subunit was significantly increased above control already 60 min after aldosterone addition, whereas beta subunit synthesis increased only 6 h after hormone addition, with no significant change for the gamma subunit. The half-lives of each subunit as assessed by 35S methionine pulse chase experiments were short (between 40 and 50 min) and were not modified by aldosterone. Taking into account the short half-life of ENaC protein and assuming that the synthesis of the alpha subunit is a limiting factor in the assembly and expression of new channels at the cell surface, it is proposed that the aldosterone regulation of sodium transport might be, in part, mediated by de novo synthesis of the channel protein. PMID- 9402083 TI - Localization of ROMK channels in the rat kidney. AB - Renal potassium secretion occurs in the distal segments of the nephron through apically located secretory potassium (SK) channels. SK may correspond to the ROMK channels cloned from rat kidney. In this study, the localization of ROMK at the cellular level in the rat kidney was examined using an affinity-purified polyclonal antibody raised against a C-terminal peptide of ROMK. The specificity of the antibody was demonstrated by immunoblots of membranes of Xenopus oocytes expressing ROMK2. Immunoblots of homogenates from rat renal outer medulla and cortex revealed predominant bands of 70 to 75 kD, which were ablated by preadsorption with an excess of peptide. These bands were specific for the rat kidney. Immunolocalization studies revealed that ROMK is expressed in specific nephron segments in both the cortex and medulla. In the cortex, ROMK was found in the apical domain of the thick ascending limb of Henle's loop, the connecting tubule, and in some, but not all, cells of cortical collecting tubules. In the medulla, expression in the apical membrane of the thick ascending limbs of Henle's loop was strong, whereas outer medullary collecting ducts were weakly stained. Expression in the thick ascending limb was also heterogeneous; some cells that expressed the Na-K-Cl cotransporter were weakly stained with the anti ROMK antibody. No staining of glomeruli, proximal tubules, or inner medullary collecting ducts was found. The localization of ROMK agrees well with the findings of SK in patch-clamp studies and supports the view that ROMK is the SK channel of the distal segments of the nephron. PMID- 9402084 TI - Potential role of luminal potassium in tubuloglomerular feedback. AB - Transport through the Na+-2Cl(-)-K+ cotransporter in the luminal membrane of macula densa cells is considered critical for tubuloglomerular feedback (TGF). Although various studies could support the importance of luminal Na+ and Cl-, the role of luminal K+ in TGF has not been thoroughly addressed. The study presented here examines this issue in nephrons with superficial glomeruli of anesthetized male Munich-Wistar-Fromter rats. Ambient Na+ concentration in early distal tubular fluid was approximately 22 mM, suggesting collection sites relatively close to the macula densa segment. First, it was found that ambient early distal tubular K+ concentration is approximately 1.3 mM, i.e., close to the K+ affinity of the Na+-2Cl(-)-K+ cotransporter in the thick ascending limb. Second, it was observed that a change in late proximal tubular flow rate, i.e., a maneuver that is known to induce a TGF response, significantly alters early distal tubular K+ concentration. Third, previous experiments failed to show an inhibition in TGF response during retrograde perfusion of the macula densa with K+-free solutions. Because of a potential K+ influx into the lumen between the perfusion site and the macula densa, however, the K+ channel blocker U37883A was added to the K+ free perfusate. TGF response was assessed as the fall in nephron filtration rate in response to retrograde perfusion of the macula densa segment from early distal tubular site. It was observed that luminal U37883A (100 microM) significantly attenuated TGF. Because adding 5 mM KCl to the perfusate restored TGF in the presence of U37883A and because the inhibitory action of U37883A on tubular K+ secretion was confirmed, the effect of U37883A on TGF was most likely caused by inhibition of K+ influx into the perfused segment, which decreased luminal K+ concentration at the macula densa. The present findings support a potential role for luminal K+ in TGF, which is in accordance with a transmission of the TGF signal across the macula densa via Na+-2Cl(-)-K+ cotransporter. PMID- 9402085 TI - A role for P-selectin in neutrophil and platelet infiltration in immune complex glomerulonephritis. AB - P-selectin is one of the key early mediators of leukocyte adhesion in inflammatory conditions. This report examines the role of P-selectin in a neutrophil- and platelet-mediated model of glomerulonephritis (the concanavalin A [con A] model). The administration of neutralizing anti-P-selectin antibody (PB 1.3) reduced the platelet influx at 10 min (P < 0.05) and was associated with a 60% reduction in the neutrophil infiltrate and a 50% reduction in the number of oxidant-producing cells at 3 h within glomeruli. No effect on glomerular monocyte macrophage accumulation was observed, and proteinuria was reduced by 20% but did not reach significance. It is concluded that P-selectin plays an important role in mediating the neutrophil and platelet accumulation in this model and likely has a role in mediating the glomerular injury. PMID- 9402086 TI - Cytokines and Fas regulate apoptosis in murine renal interstitial fibroblasts. AB - Renal fibrosis is characterized by an increased number of fibroblasts and excessive deposition of extracellular matrix. Apoptotic cell death is a physiological mechanism to limit cell numbers, and an insufficient rate of death may contribute to fibroblast accumulation. However, little is known about the regulation of renal fibroblast survival. The authors have studied the interaction of cytokines and the Fas receptor in the regulation of apoptosis of renal fibroblasts and have observed that murine renal fibroblasts express Fas and the Fas ligand. Tumor necrosis factor alpha (TNFalpha) and agonistic anti-Fas antibodies induce apoptosis of renal fibroblasts in a time- and dose-dependent manner. Serum contains survival factors for renal fibroblasts. Both serum deprivation and TNFalpha increase the sensitivity to Fas-induced death and the expression of fas mRNA and Fas receptor. By contrast, insulin-like growth factor 1 decreases apoptosis induced by both serum deprivation and Fas activation and partially prevents the increase in Fas receptor expression induced by serum deprivation. Murine renal fibroblasts express constitutively both fas ligand mRNA and cell-surface Fas ligand, but the authors could not demonstrate a role for Fas ligand in the autocrine regulation of fibroblast survival. These data suggest that Fas and other cytokines cooperate to regulate renal fibroblast apoptosis. Modulation of the Fas death-signaling pathway in renal fibroblasts could represent a new therapeutic target for renal fibrosis. PMID- 9402087 TI - Mutations in the vasopressin V2 receptor and aquaporin-2 genes in 12 families with congenital nephrogenic diabetes insipidus. AB - Congenital nephrogenic diabetes insipidus (CNDI) is a rare inherited disorder characterized by renal tubular insensitivity to the antidiuretic effect of arginine vasopressin (AVP). In a large majority of the cases, nephrogenic diabetes insipidus is an X-linked recessive disorder caused by mutations in the AVP V2 receptor gene (AVPR2). In the remaining cases, the disease is autosomal recessive or dominant and, for these patients, mutations in the aquaporin 2 gene (AQP2) have been reported. Fourteen probands belonging to 12 families were analyzed by single-strand conformational polymorphism and direct sequencing of the AVPR2 and AQP2 genes. Ten mutations of the AVPR2 gene (six previously reported mutations and four novel mutations: G107E, W193X, L43P, and 15delC) were identified. Three mutations of the AQP2 gene were also identified in two patients: the first patient is homozygous for the R85X mutation and the second is a compound heterozygote for V168 M and S216P mutations. Extrarenal responses to infusion of the strong V2 agonist 1-desamino-8-D-arginine vasopressin allowed AVPR2- and AQP2-associated forms of CNDI to be distinguished in three patients. This test also identified an unexpectedly high urinary osmolality (614 mosmol/kg) in a patient with a P322S mutation of AVPR2 gene and a mild form of CNDI. PMID- 9402088 TI - Arginine vasopressin secretion with mutants of wild-type and Brattleboro rats AVP gene. AB - Defects in peptide processing are associated with several disorders, including central diabetes insipidus (CDI). In the Brattleboro (BB) rat with CDI, the mRNA and protein of arginine vasopressin (AVP) are present in the hypothalamus, but no circulating AVP is detectable, thus suggesting a processing defect. The present study examined AVP secretion in cultured COS cells transfected with various constructs from wild-type and mutated Brattleboro AVP gene precursors. The precursor contains three exons encoding for vasopressin (VP), neurophysin (NP), and glycopeptide (GP). The Brattleboro rat has a deletion of a single base, guanine (G), in the NP coding region that leads to a frameshift, resulting in the loss of normal stop codon. The wild-type pcVP (22.0 +/- 5.2 pg/10[-2] U beta galactosidase [beta-gal]), but not the mutated BB AVP gene pcBB (1.2 +/- 0.4 pg/10[-2] U beta-gal), was associated with AVP secretion from the COS cells as measured by RIA. The wild-type AVP gene without the GP coding region was associated with AVP release greater (47.4 +/- 13.5 pg/10[-2] U beta-gal, n = 5, P < 0.05, versus pcVP) than the pcVP with intact VP, NP, and GP coding regions. However, the wild-type AVP gene with VP coding region alone was not processed and secreted. Normalizing the pcBB total length with the insertion of a stop codon at the site of the normal stop codon was not associated with AVP secretion (3.0 +/- 1.4 pg/10[-2] U beta-gal). However, insertion of a stop codon so that the pcBB length equaled the length of VP and NP coding regions of the wild type was associated with AVP secretion (13.5 +/- 4.0 pg/10[-2] U beta-gal). When a stop codon was inserted into the wild-type NP coding region at the same site as the G deletion in the pcBB, the AVP secretion was significantly lower (15.1 +/- 5.0 pg/10[-2] U beta-gal) than pcVP with VP + NP but no GP coding regions (47.4 +/- 13.5 pg/10[-2] U beta-gal, n = 5, P < 0.05). In summary, (1) both VP and intact NP, but not GP, coding regions are necessary for AVP processing and secretion; (2) decreasing the length of the NP coding region diminishes but does not abolish AVP processing and secretion; and (3) shortening of the pcBB length with a stop codon at a site comparable to wild-type VP + NP allows AVP secretion, albeit less than with wild-type gene precursor. Thus, the CDI in BB rats is caused by the G deletion in NP coding region. This defect leads to abnormalities that contribute to the abnormal AVP processing. Specifically, the frameshift and absence of a stop codon cause a mutated extended C terminus, which, along with the mutated NP, contribute to the abnormal steps of AVP processing, transport, and secretion in the BB rat. These defects no doubt impair the folding and configuration necessary for normal processing of the AVP gene precursor. PMID- 9402089 TI - Estrogen replacement during hypoalbuminemia may enhance atherosclerotic risk. AB - Estrogen replacement therapy is considered antiatherosclerotic because it reduces LDL cholesterol and fibrinogen and increases HDL cholesterol concentrations. However, exogenous estrogen is also known to increase hepatic triglyceride production. Hyperlipidemia in the nephrotic syndrome is probably due to increased lipoprotein secretion into plasma and decreased clearance of lipoprotein cholesterol and triglycerides. Previously, lipid-lowering effects of ovariectomy in analbuminemic rats were observed, suggesting that in the presence of hypoalbuminemia, estrogen replacement may have adverse effects on the lipid profile. To test this hypothesis, ovariectomized control rats and rats with Adriamycin-induced nephrotic syndrome were treated with estradiol. In ovariectomized controls, estradiol reduced plasma LDL cholesterol, apolipoprotein B, and fibrinogen and increased apolipoprotein A-I and triglycerides. Nephrotic rats were characterized by a marked decrease in plasma colloid osmotic pressure, hyperfibrinogenemia, hyperlipidemia, and stimulated hepatic fatty acid synthesis. The beneficial effects of estradiol on LDL cholesterol, apolipoprotein B, and fibrinogen found in ovariectomized controls were not present in estradiol-treated nephrotic rats. This suggests that in hypoalbuminemia, downregulation of the LDL receptor overrides putative estradiol-induced increases in LDL receptor activity. Moreover, estrogen replacement in the nephrotic syndrome doubled fatty acid synthesis and triglyceride secretion, and markedly exacerbated hypertriglyceridemia, suggesting saturation of triglyceride clearance. Thus, severe hypoalbuminemia in rats induces an atherosclerotic metabolic response that is aggravated by estrogen replacement. These findings suggest that estrogen replacement in hypoalbuminemic subjects could be contra-indicated. PMID- 9402090 TI - Verocytotoxin inhibits mitogenesis and protein synthesis in purified human glomerular mesangial cells without affecting cell viability: evidence for two distinct mechanisms. AB - Acute renal failure is one of the hallmarks of the hemolytic uremic syndrome (HUS). Infection with a verocytotoxin (VT)- or Shiga-like toxin (SLT)-producing Escherichia coli has been strongly implicated in the etiology of the epidemic form of HUS. The functional receptor for these closely related toxins appears to be a glycosphingolipid, globotriaosylceramide (Gb3). Endothelial damage in the glomeruli and arterioles of the kidney induced by VT is believed to play a crucial role in the pathogenesis of HUS. However, little information is available regarding the effects of VT on mesangial cells, which also play an important role in glomerular function. In this study, the effects of VT on human mesangial cells in vitro were investigated. Mesangial cells were enriched by collecting hillock shaped outgrowths derived from adult human glomeruli and subsequently purified by elimination of contaminating epithelial cells by immunoseparation with ulex europaeus lectin-I (UEA-I)-coated dynabeads. The obtained and subcultured mesangial cell populations were >98% pure. Their mesangial nature was established by the presence of a-smooth muscle cell actin in highly confluent cultures and the absence of cytokeratin or platelet/endothelial cell adhesion molecule-1. Mesangial cells bound VT to bands of Gb3 and a closely related glycolipid, which is similar to a glycolipid involved in the VT-dependent cytokine production in monocytes. VT did not induce the release of cytokines or chemokines in mesangial cells. In VT-susceptible cells, binding of VT to Gb3 causes cell death by the inhibition of protein synthesis. Although protein synthesis was inhibited in mesangial cells, all cells remained viable, both under basal and tumor necrosis factor-alpha-stimulated conditions. However, the marked reduction in protein synthesis may impair a proper response of the cells in conditions of increased demand of newly synthesized proteins. Furthermore, VT markedly inhibited DNA synthesis and proliferation of mesangial cells. The inhibition of mitogenesis was also found with the B-subunit of VT-1 alone, albeit to a lesser extent, without a significant effect on protein synthesis. Because the inhibition of protein synthesis involves the A-subunit, this suggests that two distinct mechanisms contribute to the effects of VT on protein synthesis and mitogenesis. Intracellular routing of VT (A- and B-subunits) may vary between cell types and result in differential effects on human mesangial cells when compared with other cell types. PMID- 9402091 TI - Apolipoprotein B, fibrinogen, HDL cholesterol, and apolipoprotein(a) phenotypes predict coronary artery disease in hemodialysis patients. AB - Patients with end-stage renal disease have a markedly elevated risk for coronary artery disease (CAD). Lipids and most lipoproteins, however, seem to be not predictive for CAD in these patients. Although there is clear evidence that lipoprotein(a) [Lp(a)] is significantly elevated in these patients, no study with a sufficiently large group of hemodialysis patients has investigated the relationship between CAD and Lp(a), as well as the genetically determined apolipoprotein(a) [apo(a)] phenotype. This cross-sectional study determines the prevalence of CAD in relation to the cardiovascular risk profile in an unselected population of 607 hemodialysis patients, of which 33% were diabetic patients. Twenty-six percent (n = 158) of all patients suffered from CAD as diagnosed by a definitive myocardial infarction (n = 102) and/or at least one stenosis >50% of a coronary artery (n = 143). In univariate analysis, several classic risk factors, including the concentration of lipids, lipoproteins, apolipoproteins, and fibrinogen, correlated with CAD. Lp(a) in patients with CAD showed only a tendency to higher levels, without reaching significance, compared with patients without CAD (26.6 +/- 30.8 mg/dl versus 22.1 +/- 30.4 mg/dl, P = 0.10). The frequency of low molecular weight apo(a) isoforms, however, was significantly greater in the group with CAD (34.8% versus 23.6%, P < 0.01). Stepwise logistic regression analysis found seven variables associated with CAD: apolipoprotein B, the low molecular weight apo(a) phenotype, male sex, age, fibrinogen, diabetes mellitus, and HDL cholesterol. The association of these variables with CAD differed depending on age. These results indicate that, besides classic risk factors such as age, sex, and diabetes mellitus, additional factors of the lipoprotein and fibrinolytic system contribute to the high prevalence of CAD in hemodialysis patients. PMID- 9402092 TI - Metabolic consequences of folate-induced reduction of hyperhomocysteinemia in uremia. AB - Plasma homocysteine, a well-recognized risk factor for cardiovascular disease, is elevated in uremic patients on hemodialysis. The authors have recently demonstrated that one consequence is the reduction in red cell membrane protein methylation levels, caused by a rise of intracellular adenosylhomocysteine, a potent inhibitor of methyltransferases. Protein methylation is involved in a repair mechanism of damaged membrane proteins, and an impairment in methylation leads to the accumulation of altered proteins. Therapy with folates, cofactors in the transformation of homocysteine to methionine, is effective in lowering plasma homocysteine. This article details a study on the metabolic effects of oral methyltetrahydrofolate, the active form of folic acid, on 14 uremic hemodialysis patients. Two months of therapy led to a significant reduction of plasma homocysteine levels, with a proportional response to pre-folate levels. In five of 13 patients with homocysteine levels above 20 microM, plasma homocysteine level was reduced to less than 15 microM. After treatment, levels of adenosylmethionine, the methyl donor in transmethylations, had significantly increased; levels of adenosylhomocysteine had increased to a smaller extent. Therefore, the ratio between the two compounds, an excellent indicator of the presence and the degree of methylation inhibition, was significantly ameliorated. Methionine plasma levels increased after treatment in all patients and were correlated with posttreatment adenosylmethionine levels. It was concluded that treatment with methyltetrahydrofolate brings the plasma homocysteine concentration back to an "acceptable" level, and the metabolic consequences are in the direction of an increase in the normal flow of transmethylations, as monitored by an increase in the [adenosylmethionine]/[adenosylhomocysteine] ratio. PMID- 9402093 TI - Total body water and body composition in chronic peritoneal dialysis patients. AB - In this investigation, total body water (TBW) in ten chronic peritoneal dialysis patients was studied by deuterium (TBW-2H), skinfold thickness (TBW-ST), Watson formula (TBW-WA), 58% of body weight (TBW-58%), and bioelectrical impedance (TBW BIA), and these results were compared with the reference oxygen18 (TBW-18O) method. We also analyzed the fat-free mass (FFM) by skinfold thickness (FFM-ST), bioelectrical impedance (FFM-BIA), oxygen18 (FFM-18O), and creatinine kinetics method (FFM-CK). In addition, resting metabolic rate was measured by indirect calorimetry. Compared with TBW-18O, TBW-58% and TBW-BIA were significantly different (P < 0.01). TBW-2H overestimated TBW-18O by 4.3%. TBW-ST and TBW-WA gave slightly greater values than TBW-18O, although these values were nonstatistically significant. The best prediction of total body water from these methods was obtained with the Watson formula. When Kt/V was calculated from these results, the values obtained were statistically greater (BIA, P < 0.001) and smaller (58% BW, P < 0.01) than those obtained with either 18O or Watson formula. The fat-free mass estimation also led to discrepant findings. Indeed, FFM-CK was significantly lower (P < 0.05) as compared with FFM-ST, FFM-BIA, or FFM-18O. Resting metabolic rate was strongly correlated with FFM estimated by skinfold thickness (r = 0.91, P < 0.001), bioelectrical impedance (r = 0.85, P < 0.005), and 18O (r = 0.77, P < 0.01), but not when fat-free mass was estimated by the creatinine kinetic method. The water content of fat-free mass estimated by skinfold thickness was found to be 69.7 +/- 6.9% in these patients, a value lower than the standard 73.2% found in healthy adults. This study confirms that there is an abnormal water distribution in chronic peritoneal dialysis patients. However, when compared with the oxygen18 reference method, the Watson formula allows a reliable estimation of Kt/V. PMID- 9402094 TI - Hyaluronan decreases peritoneal fluid absorption in peritoneal dialysis. AB - Hyaluronan, exhibiting a high resistance against water flow, acts in the tissue as a barrier against rapid changes in water content. To test whether hyaluronan has any effect on the peritoneal fluid and solute transport, and, in particular, on the peritoneal fluid absorption, a 4-h dwell study with an intraperitoneal volume marker (radiolabeled human serum albumin [RISA]) was conducted in 21 male Sprague Dawley rats (three groups, seven rats in each group). Each rat was injected intraperitoneally with 25 ml of 1.36% glucose solution alone (control group), with 0.005% hyaluronan (HA1 group), or with 0.01% hyaluronan (HA2 group). Dialysate and blood samples were taken frequently for analyses of fluid and solute (urea, glucose, and protein) transport. The intraperitoneal volume was calculated from the dilution of RISA with a correction for RISA disappearance from the peritoneal cavity. This study shows that adding hyaluronan to peritoneal dialysis solution significantly (P < 0.01) increased the net peritoneal fluid removal, mainly due to a significant decrease in the peritoneal fluid absorption rate (P < 0.01). The diffusive mass transfer coefficients for glucose, urea, and protein did not differ between the three groups. The peritoneal clearance of urea increased significantly in the two hyaluronan groups compared with the control group, due to the increased net fluid removal in the hyaluronan groups. These results suggest that intraperitoneal administration of hyaluronan during a single peritoneal dialysis exchange may significantly increase the peritoneal fluid and solute removal by decreasing peritoneal fluid absorption. PMID- 9402095 TI - Anemia in hemodialysis patients: variables affecting this outcome predictor. AB - Despite the prevalent use of recombinant human erythropoietin (rhEPO), anemia is a frequent finding in hemodialysis patients. The goal of this study was to evaluate the impact of anemia on patient survival and characterize the determinants of hematopoiesis that may be amenable to therapeutic manipulation to enhance rhEPO responsiveness and reduce death risk. Patient characteristics and laboratory data were collected for 21,899 patients receiving hemodialysis three times per week in dialysis centers throughout the United States in 1993. Hemoglobin concentrations (Hb) < or =80 g/L were associated with a twofold increase in the odds of death (odds ratio = 2.01; P = 0.001) when compared with Hb 100 to 110 g/L. No improvement in the odds of death was afforded for Hb >110 g/L. Using multiple linear regression, variables of rhEPO administration (rhEPO dose and percentage of treatments that rhEPO was administered), variables of iron status (serum iron, transferrin saturation, and ferritin), variables of nutritional status (serum albumin and creatinine concentration), and the dose of dialysis (urea reduction ratio) were found to be significantly associated with hemoglobin concentration (P < 0.001). Age, race, and gender were also found to be significantly associated with hemoglobin concentrations (P < 0.001). From this report, the following conclusions may be made. (1) Anemia may be predictive of an increased risk of mortality in some hemodialysis patients. (2) Hemoglobin concentrations > 110 g/L are not associated with further improvements in the odds of death. (3) Laboratory surrogates of iron stores, nutritional status, and the delivered dose of dialysis are predictive of hemoglobin concentration. Whether manipulation of the factors that improve anemia will also enhance the survival of patients on hemodialysis is unknown and should be evaluated by prospective, interventional studies. PMID- 9402096 TI - Evaluation of pathologic criteria for acute renal allograft rejection: reproducibility, sensitivity, and clinical correlation. AB - This study was designed to evaluate the pathologic criteria used for acute renal allograft rejection that were developed by a panel of renal pathologists participating in the Cooperative Clinical Trials in Transplantation, a National Institutes of Health-supported, multicenter research group. The panel defined three categories of acute rejection. (1) Type I: mononuclear infiltrate in > or =5% of cortex, a total of at least three tubules with tubulitis in 10 consecutive high-power fields from the most severely affected areas, and at least two of the three following features: edema, activated lymphocytes, or tubular injury. (2) Type II: arterial, or arteriolar, endothelialitis with or without the preceding features. (3) Type III: arterial fibrinoid necrosis or transmural inflammation with or without thrombosis, parenchymal necrosis, or hemorrhage. Using these criteria, and without any knowledge of the clinical course or original diagnosis, a rotating panel of three pathologists agreed with the original study pathologist's diagnosis of the presence or absence of rejection in 259 of the 286 biopsies (91%) used for this analysis (kappa = 0.80). The sensitivity to establish the diagnosis of rejection was 91% for a single core and 99% for two cores. To validate the diagnostic criteria, the thresholds for number of tubules with tubulitis and the percent infiltrate were varied, and the pathologic diagnosis was compared with the clinical course. The greatest agreement occurred with a threshold of > or =1 tubule with tubulitis and > or =5% cortex with interstitial infiltrate (91%). Clinically severe rejection episodes were correlated with the type of rejection (type I, odds ratio [OR] 6.2; type II, OR 37.9). Type II rejection was more likely to be clinically severe than type I (OR 6.1). Analysis of other individual pathologic features revealed a correlation with clinical severity for endothelialitis (OR 13.2), interstitial hemorrhage (OR 13.2), and the presence of glomerulitis (OR 3.7) (all P < 0.05). The extent of tubulitis or of the interstitial infiltrate did not correlate with severity (P > 0.05). It is concluded that these criteria are simple, reproducible, and clinically relevant. These data should lead to further refinement of the diagnostic systems for renal allograft rejection. PMID- 9402097 TI - Family history of end-stage renal disease among incident dialysis patients. AB - As part of a larger study of genetic risk factors for the occurrence of renal failure, the prevalence of a family history of end-stage renal disease (ESRD) in first- and second-degree relatives of all incident dialysis patients treated in Georgia, North Carolina, and South Carolina (ESRD Network 6) in 1994 was ascertained. Family histories were obtained from 4365 dialysis patients (83% of those eligible), and 856 (20%) reported having a family history of ESRD. Among race-sex groups, 14.1% of Caucasian men, 14.6% of Caucasian women, 22.9% of African-American men, and 23.9% of African-American women reported a first- or second-degree relative with ESRD (P = 0.001). The prevalence of relatives with ESRD varied by the reported etiology: 22.2% in diabetes mellitus; 18.9% in hypertension, 22.7% in glomerulonephritis; and 13.0% of other etiologies (P = 0.001). Patient characteristics independently associated with family history of ESRD included race, younger age, higher levels of education, and etiology of ESRD. In this report, it is concluded that a large proportion of incident ESRD cases have close relatives with ESRD in whom preventive actions might be directed. Genetic analyses in multiply affected families may identify the inherited factors contributing to progressive renal failure. PMID- 9402098 TI - Blood flow limitations of solute transport across the visceral peritoneum. AB - In a previous study, no limitations to urea transfer across the parietal peritoneum were demonstrated with decreases in local blood flow of 70%. It was hypothesized that the visceral peritoneum would have similar characteristics. To address this problem at the tissue level, diffusion chambers were affixed to the serosal side of the stomach, cecum, or liver of anesthetized rats (n = 6 each tissue), and solutions containing 14C urea were placed in the chamber. During each experiment, the local chamber blood flow was measured with laser Doppler flowmetry, and, simultaneously, the disappearance of the tracer versus time was determined under three conditions: control, after 60 to 70% blood flow reduction, and postmortem (flow = 0). The results showed no difference in the urea mass transfer coefficient (MTC; mean +/- SEM; cm/min x 10[3]) between control and blood flow reduction for the stomach (4.0 +/- 0.4 versus 3.6 +/- 0.3) or for the cecum (4.6 +/- 0.3 versus 4.0 +/- 0.3). However, the MTC was significantly decreased by local blood flow reduction in the liver (5.4 +/- 0.2 versus 2.6 +/- 0.2). Postmortem data demonstrated significant reductions in the MTC with blood flow equal to zero. It is concluded that a 60 to 70% blood flow reduction from control values does not limit solute transperitoneal transfer in the hollow viscera but causes significant changes in the mass transfer across the liver surface. Because the liver makes up only a small portion of the effective exchange area, overall transperitoneal solute transfer should not be greatly affected by significant decreases in blood flow. PMID- 9402099 TI - Congenital nephrogenic diabetes insipidus. PMID- 9402100 TI - The artificial kidney: a dialyser with a great area. 1944. PMID- 9402101 TI - Glomerulomegaly and proteinuria in a patient with idiopathic pulmonary hypertension. AB - Glomerulomegaly is a histologic finding present in idiopathic pulmonary hypertension, congenital cyanotic heart disease, morbid obesity associated with sleep apnea syndrome, sickle cell disease, and polycythemic states. This study examines the case of a 34-yr-old woman with idiopathic pulmonary artery hypertension who presented with nephrotic-range proteinuria. Kidney biopsy revealed enlarged glomeruli with mesangial-proliferative glomerulonephritis. A review of the pertinent literature and a discussion of the proposed pathophysiologic mechanisms leading to glomerulomegaly are presented. PMID- 9402103 TI - Transforming growth factor-beta1 in adult human microglia and its stimulated production by interleukin-1. AB - Ameboid microglia express human immunodeficiency virus 1 (HIV-1) more frequently than do ramified microglia. These two microglial subtypes might also differ in the frequency with which they express transforming growth factor-beta1 (TGF beta1), a cytokine that regulates HIV-1 expression in monocytes. Results described here show that ameboid and ramified microglia express TGF-beta1. In brain tissues from HIV-1-infected individuals as compared with seronegative controls, ameboid rather than ramified microglia more frequently expressed TGF beta1. Ameboid microglia, isolated and cultured from postmortem adult human brain more frequently expressed TGF-beta1 in presence of interleukin-1(IL-1), a cytokine that is elevated in brains of HIV-1-infected individuals when compared with seronegative controls. The stimulation of TGF-beta1 by IL-1 was dose and time dependent, occurring with ameboid microglia isolated from either frontal cortex or globus pallidus but not midbrain pons. Ameboid microglia are similar to the RCA-1-positive cells that form clusters, called microglial nodules, in the brain of HIV-1-infected individuals. Pathologic conditions, such as disseminated microglial nodules, are associated with HIV-1 encephalitis, direct infection of the brain, and moderate to severe neurologic impairment. TGF-beta1 expression in ameboid microglia may play a role in HIV-1 neuropathogenesis. PMID- 9402102 TI - Subacute bacterial endocarditis masquerading as type III essential mixed cryoglobulinemia. AB - An adult man presented severely ill with vasculitis of his lower extremities and with impaired kidney function. After detailed evaluation at a local hospital, a diagnosis of essential type III cryoglobulinemia was made. High-dose steroid and cyclophosphamide therapy was begun. The patient improved dramatically. However, 6 wk later when his steroid dose was reduced to 30 mg daily, vasculitis recurred. Intensifying his immunosuppressive therapy only worsened his condition. He was than transferred to the Ohio State University Medical Center for consideration for plasmapheresis for the presumed essential type III cryoglobulinemia. However, our evaluation showed that he had bacterial endocarditis causing his type III cryoglobulinemia. When immunosuppressive drugs were stopped and antibiotics were begun, his condition resolved completely. This case illustrates the difficulty of identifying infectious causes of cryoglobulinemia and emphasizes that an initial, highly favorable response of vasculitis to immunosuppressive therapy does not exclude an infectious cause for the vasculitis. PMID- 9402104 TI - Activity and tolerance of a continuous subcutaneous infusion of interferon alpha2b in patients with chronic hepatitis C. AB - We administered interferon-alpha2b (IFN-alpha2b) by continuous subcutaneous infusion (60,000 IU/h, or 10 million IU/week) over 3 months to 7 patients with chronic hepatitis C. All had previously responded, as assessed by normalization of transaminases to the same dose of IFN administered by intermittent injection over 6 months, but had relapsed after cessation of therapy. The continuous infusion was tolerated well at the site of infusion, and the systemic side effects were similar in type but were lesser in intensity than with intermittent dosage. Four of 7 subjects had normalization of transaminase at the end of week 12 of therapy. Serum HCV RNA and HCV by PCR decreased with treatment, and there was a prompt and sustained increase in serum beta2-microglobulin and of 2', 5' OAS activity. The level of the latter appeared to correlate with response of the transaminase. Serum IFN concentrations were low but detectable throughout therapy. After stopping IFN administration, the transaminases in responders increased again to pretreatment levels. PMID- 9402105 TI - Cytokine modulatable signalling through macrophage HLA class II. 1. IFN-gamma upregulates the efficiency of Ca2+ mobilization in response to ligation of macrophage HLA-DP. AB - The human macrophage line 2MAC, established recently from peripheral blood, expresses a number of lineage-specific markers as well as a broad array of intercellular adhesion molecules, including high levels of HLA class I and class II. We have presented evidence elsewhere that 2MAC can be productively applied to the study of signal transduction through macrophage HLA class II. Namely, we showed that ligation of 2MAC HLA class II, but not HLA class I, by monoclonal antibody (mAb) elicits an increase in free cytoplasmic Ca2+ concentration [Ca2+]i. Moreover, this Ca2+ flux appears to be functionally relevant: ligation of HLA-DR, but not HLA class I, by mAb results in the Ca2+ mobilization-dependent induction of tissue factor, the high-affinity cellular receptor for factor VII/VIIa. Here we show that 2MAC is uniquely valuable for addressing the efficiency of signal transduction through HLA class II. Namely, we show here that prior culture of 2MAC cells with interferon-gamma (IFN-gamma) profoundly upregulates subsequent Ca2+ mobilization in response to ligation of HLA-DP in the absence of increased cell surface HLA-DP expression. Because IFN-gamma has no effect on 2MAC HLA-DP expression, IFN-gamma must upregulate Ca2+ mobilization by increasing the efficiency of signal transduction through HLA class II (HLA-DP), by targeting some other component of the macrophage HLA class II signalling pathway. PMID- 9402106 TI - Synergistic antitumor effects of a combination of interferon and tamoxifen on estrogen receptor-positive and receptor-negative human tumor cell lines in vivo and in vitro. AB - Solid tumors are relatively resistant to growth inhibition by interferons (IFNs). To enhance sensitivity, we assessed combinations of IFNs with tamoxifen in estrogen receptor-positive (ER-positive) and ER-negative human tumor xenografts. In nude mice, the growth of MCF-7 human breast tumors (ER-positive) and NIH-OVCAR 3 ovarian tumors (functionally ER-negative) was suppressed completely when tamoxifen and IFN-alpha or IFN-beta was started 2 days after tumor inoculation. Established, 6-week-old MCF-7 and NIH-OVCAR-3 tumors regressed when treated with the combination of IFN-beta and tamoxifen but not with single-agent therapy. Treatment with the combination also resulted in an augmented antitumor response in vivo in an ER-negative breast tumor (MDA-MB-231), a colon carcinoma (HT-29), and a melanoma (SK-MEL-1). Antiproliferative studies in vitro suggested that growth of both MCF-7 and NIH-OVCAR-3 cells was inhibited to a greater degree by combination treatment with human IFN-alpha and tamoxifen or IFN-beta and tamoxifen compared with single agents. Median effect analysis defined synergy. Four ER-negative carcinomas (MDA-MB-231, MDA-MB-468, BT-20, and HT-29) also exhibited synergistic growth inhibition in response to the drug combination. The response of these four cell lines was particularly striking. Tamoxifen as a single agent had little effect (up to 2.0 microM) but caused enhanced antiproliferative activity when added to IFN-beta. Sequential treatment of MCF-7 cells in vitro with tamoxifen followed by IFN-beta was more effective at inhibiting growth than treatment with IFN-beta followed by tamoxifen, suggesting that tamoxifen modulated the anticellular response to IFN-beta rather than the converse. Similar results were obtained with IFN-alpha. Cell cycle analysis indicated that 7 days of exposure to the combination resulted in MCF-7 cell fragmentation and death. Together with our recent studies demonstrating enhancement of IFN-stimulated gene expression (ISG) by tamoxifen pretreatment in IFN-resistant cells, these data suggest that combination treatment with tamoxifen and IFNs may increase ISG expression in IFN-resistant tumors, leading to augmented antitumor effects. These effects appear to be independent of ER expression. PMID- 9402107 TI - Lack of effect of different cytokines on expression of membrane-bound regulators of complement activity on human uveal melanoma cells. AB - Tumor cells are protected from antibody-dependent complement-mediated lysis by membrane-bound regulators of complement activation (m-RCA). m-RCA are expressed on uveal melanoma cells. We determined whether cytokine treatment affects expression of m-RCA on these cells in vitro. m-RCA expression on uveal melanoma cell lines was studied by flow cytometry, using monoclonal antibodies directed against CD46, CD55, and CD59. Cytokines studied were interferon-alpha (IFN alpha), IFN-gamma, interleukin-1B (IL-1B), IL-12, and tumor necrosis factor-alpha (TNF-alpha). All three m-RCA were expressed on the uveal melanoma cell lines (CD59>>CD46>CD55), although in variable amounts. With a few exceptions, the cytokines had no effect on m-RCA expression. CD55 expression was not influenced by any of the cytokines. IFN-gamma downregulated expression of CD46 on one cell line (p < 0.01). TNF-alpha upregulated CD59 expression on two of the five cell lines (p < 0.012 and p < 0.001, respectively), which effect was dose dependent. IFN-alpha, IFN-gamma, IL1-beta, IL12, and TNF-alpha had limited effects on m-RCA expression on uveal melanoma cells in vitro. PMID- 9402108 TI - Interferon-induced growth arrest is mediated by membrane structural changes. AB - Interferon-gamma (IFN-gamma) is an immunomodulator shown to augment the expression of major histocompatibility (MHC) class I/class II antigens on the cell surface. In previous studies, we have demonstrated that the enhanced expression of these antigens on the cell surface is in part due to IFN-gamma mediated abrogation of antigen shedding. In this study, we demonstrate that IFN gamma induces structural changes in the cell membrane by altering the cholesterol/phospholipid ratio. Furthermore, such changes not only mediate enhanced expression of antigen on the cell surface but may drive the cells to growth arrest and apoptosis. These results were obtained by employing x-ray diffraction, electron microscopy, and DNA analysis. PMID- 9402109 TI - Importance of pretreatment viral load and monitoring of serum hepatitis C virus RNA in predicting responses to interferon-alpha2a treatment of chronic hepatitis C. Hanshin Chronic Hepatitis C Study Group. AB - The aim of this study was to determine what factors correlate with a favorable response to interferon-alpha2a (IFN-alpha2a) treatment in chronic hepatitis C. Fifty patients with chronic hepatitis C who received a 26-week treatment with IFN alpha2a (474 million units in total) were assessed for pretreatment parameters and biochemical and virologic events during the treatment. According to biochemical and virologic responses to the treatment, 16 patients (32%) were categorized as sustained complete responders (SR), 13 (26%) as initial complete responders (IR), and 21 (42%) as nonresponders (NR). By multivariate analysis, a low viremia level was the only independent predictor of SR among pretreatment parameters (p = 0.0088). The percentage of patients showing hepatitis C virus RNA negativity at 2 and 12 weeks of treatment was significantly higher in SR (94% and 100%, respectively) or IR (69% and 92%, respectively) than in NR (14% and 33%, respectively) (p = 0.001). In contrast, the normalization of serum alanine aminotransferase levels at both time points failed to differentiate among SR, IR, and NR. These results indicate that monitoring of serum hepatitis C virus RNA at an appropriate time during treatment in addition to determination of pretreatment viral load is important in predicting responses to IFN-alpha2a treatment. PMID- 9402110 TI - Cloning and expression of the cDNA for canine interleukin-12. AB - We cloned the canine interleukin-12 (IL-12) subunit cDNA. Canine IL-12 exhibited sequence homology to the known sequences of human, mouse, and bovine genes at nucleotide and amino acid levels. Cotransfection of the p35 and p40 subunits of canine IL-12 cDNA clones into COS-1 cells resulted in the secretion of IL-12, which supported proliferation of the stimulated canine lymphocytes, promoted induction of canine interferon-gamma (IFN-gamma) from canine lymphocytes, and showed antitumor effect in vitro. The cloned canine IL-12 will be useful for canine therapeutic applications. PMID- 9402111 TI - Clinical utility of endoscopic retrograde cholangiopancreatography. AB - BACKGROUND: ERCP is a frequently performed procedure, but its necessity for diagnosis and ability to change management plans are unclear in many cases. METHODS: We prospectively evaluated diagnosis, certainty of diagnosis, and management recommendations, both before and after ERCP, as well as therapeutic maneuvers performed during ERCP, in unselected patients undergoing this procedure. RESULTS: ERCP procedures (1341) were studied at a university hospital, an ERCP referral center, and two community hospitals. Among patients undergoing first-time ERCP, the preceding clinical diagnosis was correct for 64% of those predicted to have bile duct stones, 86% to 89% of those given other biliary diagnoses, and 88% predicted to be normal. In 35% of cases, diagnostic confidence improved substantially after ERCP. Endoscopic therapy was successfully completed in 51%. After ERCP, plans for other invasive procedures changed in 82%: percutaneous biliary studies and open surgical procedures were recommended less often and laparoscopic cholecystectomy more often. Endoscopic therapy and overall clinical utility were most common in patients with cholangitis, jaundice, or bile leaks. CONCLUSIONS: ERCP is particularly helpful for diagnosis of bile duct stones but is less likely to change other diagnoses. The endoscopic therapy commonly carried out during ERCP often changes the treatment plan, leading to fewer surgical and percutaneous interventions in general, but more laparoscopic cholecystectomies. PMID- 9402112 TI - Application of magnifying chromoscopy for the assessment of severity in patients with mild to moderate ulcerative colitis. AB - BACKGROUND: A magnifying colonoscope that enables high-power observation of the colorectal mucosa has been recently developed. The aim of this study was to investigate the value of the magnifying instrument in determining the severity of ulcerative colitis. METHODS: Magnifying colonoscopy was performed in 41 patients with ulcerative colitis, and the findings in the rectum were graded according to network pattern and cryptal openings. These findings were correlated with endoscopic, clinical, and histologic severity of the disease. RESULTS: Magnifying colonoscopy did not detect network pattern in 37% and cryptal opening in 24% of the subjects. The clinical, endoscopic, and histologic grades of activity were not different between groups divided by the presence or absence of each finding. However, when the two features were coupled, patients with visible network pattern and cryptal opening had a lower clinical activity index and lower grade of histologic inflammation than those in whom both findings could not be visualized. CONCLUSIONS: Observation under magnifying colonoscopy can be another clue to determining the severity of disease in patients with ulcerative colitis. PMID- 9402113 TI - Endoscopic ultrasonography in the diagnosis of submucosal lesions of the large intestine. AB - BACKGROUND: Although reports of colorectal submucosal tumors have increased since the development of endoscopic examinations, precise diagnosis of these lesions remains difficult. In this study, we evaluated the usefulness of endoscopic ultrasonography (EUS) in the diagnosis of submucosal lesions of the large intestine. METHODS: From September 1989 to June 1996, EUS was performed in 46 patients who were suspected to have submucosal lesions by barium enema or colonoscopy. Twenty-seven of the 46 cases were confirmed histologically by endoscopic or surgical resection, and their ultrasonographic images were compared with resection specimens. RESULTS: Lipomas (n = 15) were visualized as hyperechoic masses and lymphangiomas (n = 9) visualized as cystic lesions with septal structures as characteristic findings. The EUS images of leiomyomas (n = 6), leiomyosarcomas (n = 3), and enteric endometriosis (n = 7) were all hypoechoic masses in the fourth layer. Leiomyosarcomas tended to be larger and more inhomogeneous than leiomyomas. Enteric endometriosis was shaped like a spindle or a half-moon, and myogenic tumors were lobulated when the lesions were large. Recurrences of colorectal carcinoma (n = 3), malignant lymphomas (n = 2), and an appendiceal mucocele (n = 1) were examined. CONCLUSIONS: EUS is useful in the diagnosis of submucosal lesions of the large intestine because it provides precise information about these lesions. PMID- 9402114 TI - Endosonographic imaging of pancreatic pseudocysts before endoscopic transmural drainage. AB - BACKGROUND: Endoscopic drainage of pancreatic pseudocysts has become an established alternative to surgery. We performed endosonography before endoscopic drainage to find out whether detailed anatomic information would help in the selection of appropriate candidates and result in a reduction of complications. PATIENTS AND METHODS: Between April 1992 and July 1995 endosonography was performed in 32 patients, referred for endoscopic pseudocyst drainage, to determine the minimal distance between the pseudocyst and the gut, to identify interposed vascular structures, and to determine the optimal site for drainage. RESULTS: Endosonography failed to identify a pseudocyst in 3 patients and in 2 patients the lesion was inconsistent with a pseudocyst. In 7 patients transmural drainage was considered inappropriate: in 4 the distance between the gut and the cyst was too large, in 2 varices were present between the cyst and the gut, and in 1 patient normal pancreatic parenchyma was present between the cyst and the gut. In 20 patients endosonography was followed by ERCP, and in 19 endoscopic drainage was attempted. Transmural drainage was successful in 16 patients. Endosonography changed management in 37.5% of the patients. CONCLUSION: Endosonography provides essential information prior to endoscopic drainage of pseudocysts, leading to a change in therapy in one third of patients. PMID- 9402115 TI - Incidence and clinical findings of benign, inflammatory disease in patients resected for presumed pancreatic head cancer. AB - BACKGROUND: The differentiation between cancer and benign disease in the pancreatic head is difficult. The aim of this study was to examine common features in a group of patients that had undergone pancreatoduodenectomy for a benign, inflammatory lesion misdiagnosed as pancreatic head cancer. METHODS: Among 220 pancreatoduodenectomies performed on the suspiscion of pancreatic head cancer, an inflammatory lesion in the pancreas or distal common bile duct was diagnosed in 14 patients (6%). Of these patients, all preoperative clinical information and radiologic images (ultrasound, endoscopic retrograde cholangio pancreaticography [ERCP]) were critically reassessed. For each examination, the suspicion of cancer was scored on a 0/+/++ scale. RESULTS: Clinical presentation (pain, weight loss, jaundice) raised a suspicion of cancer in 12 patients. On ultrasound, a tumor (mean size: 2.8 cm) was found in the pancreatic head in 13 patients; 12 of 14 ultrasound examinations raised a suspicion of cancer. ERCP showed a distal common bile duct stenosis (length: 1 to 4 cm), stenosis of the pancreatic duct (length: 1 to 5 cm), or a "double duct" stenosis, suspicious for cancer in 13 evaluable patients. The overall index of suspicion was + in seven patients and ++ in seven patients, confirming the initial interpretation of preoperative data. CONCLUSION: When undertaking pancreatoduodenectomy for a suspicious lesion in the pancreatic head, it is necessary to expect at least a 5% chance of resecting a benign, inflammatory lesion masquerading as cancer. PMID- 9402116 TI - Potential risks and artifacts of magnetic resonance imaging of self-expandable esophageal stents. AB - BACKGROUND: Several types of coated and uncoated self-expandable stents composed of various metals are available for palliation of malignant esophageal stenoses and fistulas. We encountered poor visualization of the mediastinum and skeletal axis with magnetic resonance imaging in a patient with a Gianturco self expandable stent. METHODS: To evaluate potential problems, such as stent migration in the magnetic field of the magnetic resonance scanner (MRI) and artifacts in the images, we studied four types of expandable stents: the Ultraflex (titanium alloy), the covered Wall stent (nitinol), the Gianturco stent (Cook), and the modified Gianturco stent (Song)-the last two being made of stainless steel. RESULTS: An appreciable attraction force and torque, as measured ex vivo by suspending the stent in a Perspex device, was found for both Gianturco stents. In particular, the Gianturco (Cook) stent is pulled toward the head with a force of 7g; however, it is uncertain whether this is a potential risk for dislodgement. In addition, gross artifacts in the images, studied by putting the stents in a cylindric phantom filled with a diluted gadolinium solution, made the magnetic resonance useless for imaging in a wide area around the Gianturco stents of stainless steel. No problems with magnetic resonance image quality were encountered for the titanium-based Ultraflex and Wall stents. CONCLUSIONS: Specific information on the type of stent is necessary before a magnetic resonance imaging examination is planned. PMID- 9402117 TI - Gastrointestinal bleeding in cirrhotic patients with hepatocellular carcinoma undergoing intrahepatic artery chemotherapy. AB - BACKGROUND: Hepatocellular carcinoma in cirrhotic patients increases the risk of variceal bleeding. We sought to characterize bleeding in a cirrhotic patient population undergoing intrahepatic artery chemotherapy for hepatocellular carcinoma and to determine the possible influence of this treatment on gastrointestinal bleeding. METHODS: We retrospectively reviewed 179 patients with hepatocellular carcinoma who underwent intrahepatic artery doxorubicin and cis platinum chemotherapy to determine the incidence of gastrointestinal bleeding and compared them with 434 hepatocellular carcinoma historic controls not undergoing regional chemotherapy. RESULTS: Of the 179 patients, 27 patients (15.1%) developed upper gastrointestinal bleeding over a mean follow-up of 15.2 months; 18 of the 27 (66.7%) bled from a variceal source and 9 (33.3%) bled from a nonvariceal source: ulcer (n = 6), gastropathy (n = 1), Mallory-Weiss (n = 1), erosive gastritis (n = 1). Twenty-one patients developed bleeding after initiation of chemotherapy (14 variceal and 7 nonvariceal). The number of chemotherapy sessions among patients with variceal and nonvariceal bleeding was similar (2.1 +/- 0.4 and 4.0 +/- 1.2; p = Not significant). Patients with variceal and nonvariceal bleeding were comparable with respect to Child-Pugh classification, pTNM stage, age, time to bleeding, and gender. CONCLUSIONS: Regional intra-arterial chemotherapy for hepatocellular carcinoma is associated with a low risk of variceal bleeding. Nonvariceal sources of upper gastrointestinal bleeding in this population account for a significant component of bleeding episodes. PMID- 9402119 TI - Randomized controlled trial of rectal tube placement for the management of abdominal distension following colonoscopy. AB - BACKGROUND: Traditionally, endoscopists have sought to provide maximum comfort for patients undergoing colonoscopy but may have been less concerned with the level of patient discomfort following the procedure. The aim of this study was to examine the effectiveness of rectal tube placement for abdominal decompression following colonoscopy in an effort to limit patient discomfort. METHODS: We conducted a prospective, single-blind, randomized controlled trial in 67 consecutive men undergoing elective colonoscopy. At the end of the procedure, patients were randomized to rectal tube placement or standard management without tube placement. Patients were evaluated by standardized criteria 30 minutes after completion of colonoscopy, and again 24 hours later. RESULTS: Thirty patients were randomized to rectal tube placement and 37 served as controls. The two groups were well matched with respect to age, duration of colonoscopy, quality of bowel preparation, prevalence of diverticulosis, frequency of polypectomy, and degree of difficulty in colonoscopy. At 30 minutes after colonoscopy, patients' overall satisfaction rating (mean +/- SD) from a 10-point scale was 9.1 +/- 3.2 in treated patients and 5.7 +/- 3.9 in controls (p < 0.05; 2-tailed unpaired t test). CONCLUSION: Placement of a rectal tube at the conclusion of colonoscopy reduces patient discomfort, and improves satisfaction. PMID- 9402118 TI - Prospective randomized comparative study of bipolar electrocoagulation versus heater probe for treatment of chronically bleeding internal hemorrhoids. AB - BACKGROUND: Our purpose was to compare the efficacy, complications, failure rates, and crossovers of heater and bipolar probe treatments of chronically bleeding internal hemorrhoids. METHODS: Eighty-one patients (31 female, 50 male) with mean age of 53 years had large (grade 2 to 3) internal hemorrhoids with bleeding for a mean of 12 years, had failed medical management, and were randomized in a prospective study of anoscopic treatments to heater versus bipolar probes. Failure was defined as a major complication or failure to reduce the size of all internal hemorrhoids with three or more treatments. RESULTS: With similar background variables and no difference in treatment times, rectal bleeding and other symptoms were controlled in a shorter time with the heater probe than with the bipolar probe (77 versus 121 days). Five complications (fissures, bleeding, or rectal spasm) occurred with the bipolar probe, and two occurred with the heater probe. The heater probe caused more pain during treatments but had significantly fewer failures and crossovers. CONCLUSIONS: For patients who had failed medical management of chronically bleeding internal hemorrhoids, the techniques and complications of heater and bipolar probes were similar, but pain was more common, failures and crossovers were less frequent, and the time to symptom relief was shorter with the heater probe than with the bipolar probe. PMID- 9402120 TI - Segmental balloon cytology for preoperative localization of in situ pancreatic cancer. PMID- 9402121 TI - Combined endoscopic and laparoscopic management of chronic gastric volvulus. PMID- 9402122 TI - Survival with early diagnosis of invasive gastric mucormycosis in a heart transplant patient. PMID- 9402123 TI - Clinical utility of percutaneous transhepatic cholangioscopy in defining tumor extent: a case of mucin-producing bile duct carcinoma originating in the left caudate lobe. PMID- 9402124 TI - Acute lower gastrointestinal bleeding from the appendix. PMID- 9402125 TI - A case of lipoma of the terminal ileum treated by endoscopic removal. PMID- 9402126 TI - Endoscopic repair by clipping of iatrogenic colonic perforation. PMID- 9402127 TI - In comes outcomes. PMID- 9402128 TI - The new endoscopy of ulcerative colitis. PMID- 9402129 TI - Tapered-tip, triple-lumen papillotome/cannula facilitates cannulation yet accepts standard guidewires. PMID- 9402130 TI - Rapid retrieval of small resected polyps. PMID- 9402131 TI - "Pull" or "push" PEG: the reinsertion of the gastroscope is often unnecessary. PMID- 9402132 TI - PEG/PEJ tube placement. PMID- 9402133 TI - Laparoscopically assisted panenteroscopy for gastrointestinal bleeding of obscure origin. PMID- 9402134 TI - The Norwegian guidelines for surveillance after polypectomy: 10-year intervals. PMID- 9402135 TI - Surgical biliary bypass for benign and malignant extrahepatic biliary obstruction. PMID- 9402136 TI - Gastrointestinal endoscopy in Israel. PMID- 9402137 TI - Endoscopy services in Malaysia--a surgical bias. PMID- 9402138 TI - The T-cell receptor as immunoglobulin: paradigm regained. AB - The quest to determine the molecular nature of T-lymphocyte receptors for antigen was a "holy grail" to immunologists for over 25 years. This paper updates a review written 15 years ago (Marchalonis JJ, Hunt JC. Proc Soc Exp Biol Med 171:127-145, 1982), which proposed that "these molecules apparently do not bear determinants specified by the major histocompatibility complex, but express Ig related variable regions and constant regions unique to T-cell products." We review subsequent contributions from molecular biology, protein chemistry, peptide immunochemistry, and structural biology establishing that T-cell receptors (TCRs) are members of the immunoglobulin family restricted to T cells that share 3-dimensional structural features, sequence homology, antigenic cross reactivity, and common mechanisms of diversification with conventional immunoglobulins. These molecules and their light- and heavy-chain siblings appeared contemporaneously in vertebrate evolution with the emergence of sharks. We illustrate how extrapolation of concepts from immunoglobulin to T-cell receptors has aided in the understanding of these often enigmatic molecules, and, conversely, how concepts derived for T-cell receptors such as the role of "superantigens" can be directly applied to conventional immunoglobulins. A second precept that follows from the symmetry of the combining sites of Igs and TCRs is that MHC-restricted antibodies should exist. Such molecules have in fact been reported, and the x-ray crystallography for T-cell receptors suggests that the combining sites recognizing simultaneously MHC and peptide epitopes resemble the combining sites of antibodies directed against protein determinants. Additional immunoglobulin molecules of nonmammalian species have been detected and characterized based upon conserved homology to TCR and Igs, and it is anticipated that further study will enable the identification of more antigen-specific members of the family in mammals as well. PMID- 9402140 TI - Erythropoietin: physiologic and pharmacologic aspects. AB - The purpose of this review is to give an update of the recent progress in research on erythropoietin (Epo), the hormone that regulates red blood cell production. Epo is a glycoprotein with a molecular mass of approx 30 kDa, which circulates in plasma of the human with 165 amino acids with three N-linked and one O-linked acidic oligosaccharide side chains in the molecule. Both the alpha (39% CHO) and beta (24% CHO) forms are available for clinical use, and there does not appear to be any difference in the pharmacokinetics of these two forms of Epo. Radioimmunoassays and enzyme-linked immunoabsorbant (ELISA) assays are available in a kit form. Serum levels of Epo in normal human subjects range between 1 and 27 mmu/ml or approx 5 pmol/l. It seems clear that the cells in the adult mammalian kidney which produce Epo are the interstitial cells in the peritubular capillary bed and the perivenous hepatocytes in the liver. Expression of the human Epo gene sequences that direct expression in the kidney are located 6-14 kilobases 5' to the gene; whereas the sequences that control hepatocyte specific expression are located within 0.7 KS to the 3'-flanking region and 0.5 KS to the 5'-flanking region. The signal transduction pathways postulated to be involved in the expression of Epo are: kinases A, G and C; both a constitutive factor and a second hypoxia-inducible factor-1 (HIF-1) located in the 5' end of an hypoxia inducible enhancer region of the Epo gene; and reactive oxygen species. The primary target cell in the bone marrow acted on by Epo is the colony forming unit erythroid (CFU-E) which has the highest number of Epo receptors. It has been postulated that Epo decreases the rate which Epo-dependent progenitor cells undergo programed cell death (apoptosis). There are two major signal transduction pathways activated by the Epo receptor: the JAK2-STAT5 pathway and the ras pathway. Both pathways involve tyrosine phosphorylation. The approved clinical uses of Epo are the anemias associated with end-stage renal disease, cancer chemotherapeutic agents, and patients with HIV infection receiving AZT. Other anemias reported to respond to Epo therapy are anemia of prematurity, rheumatoid arthritis, and myelodysplasia. Other uses of Epo under investigation are in perioperative surgery and preoperative autologous blood donation. PMID- 9402139 TI - Insulin-like growth factor binding protein-1: recent findings and new directions. AB - In 1988, insulin-like growth factor-binding protein-1 (IGFBP-1) became the first characterized member of a group of structurally related soluble proteins which specifically bind and modulate the actions of the IGFs. Since then, a wealth of information has accumulated regarding the physiology of this dynamic serum protein. In this review, we update our 1993 summary (Lee PDK et al. Proc Soc Exp Biol Med 204:4-29) of the status of IGFBP-1 research. The IGFBP-1 protein sequence contains 12 N-terminal and 6 C-terminal cysteine residues which are conserved in other mammalian IGFBP-1 sequences and amongst other IGFBPs; both of the cysteine-rich regions are required for optimal IGF binding. The nonconserved IGFBP-1 midregion may act as both a hinge which defines ligand binding characteristics and as a specific target for protease activity. Integrin-binding and phosphorylation sites within the IGFBP-1 sequence have functional significance in vitro, but their physiologic relevance in vivo have not been defined. The human IGFBP-1 and IGFBP-3 genes are contiguous and located in close proximity to the homeobox A (HOXA) gene cluster on chromosome 7. The other IGFBP genes, located on chromosomes 2, 12, and 17, are also associated with HOX clusters, suggesting evolutionary linkage of the IGFBP and HOX gene families. Similarities between the hIGFBP-1 and phosphoenolpyruvate kinase (PEPCK) promoters, including regions conferring insulin, glucocorticoid, and cyclic adenosine-monophosphate responses, are consistent with our previous hypothesis that IGFBP-1 is involved in regulation of glucose metabolism. The tissue-specific patterns of IGFBP-1 gene expression in liver, kidney, decidua, and ovary may be due to stimulation of IGFBP-1 transcription by hepatic nuclear factor 1 (HNF1) proteins. Clinical and basic studies of IGFBP-1 physiology have been aided by several recently developed assay methods. Numerous investigations have confirmed that insulin, via inhibition of IGFBP-1 transcription, is the primary determinant of IGFBP-1 expression both in vitro and in vivo. IGF-I and IGF-II also have specific inhibitory effects on IGFBP-1 expression. Glucocorticoids and cAMP stimulate IGFBP-1 transcription, but these effects are observed only in conditions of low or absent insulin effect. Other stimulants of IGFBP-1 expression include thyroid hormones and epidermal growth factor. Phorbol ester stimulation of IGFBP-1 expression can supersede the effects of insulin in vitro;however, the mechanism and in vivo correlates of this effect have not been determined. Cytokines and, perhaps, growth hormones may affect IGFBP-1 expression, perhaps by altering the regulatory actions of insulin; this effect may have important clinical relevance. IGFBP-1 expression is upregulated in liver and (nonhuman) kidney during postinjury regeneration. The IGF-inhibitory actions of IGFBP-1 has been confirmed by numerous in vitro studies and several in vivo animal investigations, including administration of recombinant IGFBP-1 and IGFBP 1 transgenic models. IGFBP-1 has been shown to inhibit somatic linear growth, weight gain, tissue growth, and glucose metabolism. Moreover, IGFBP-1 appears to be a primary determinant of free IGF-I levels in serum. Excess levels of IGFBP-1 may contribute to growth failure in intrauterine growth restriction and in pediatric chronic renal failure, while low IGFBP-1 levels are associated with obesity and with cardiovascular risk factors in insulin resistance syndromes. Serum IGFBP-1 measurements may be useful biochemical marker in these pathologic conditions. IGFBP-1 is expressed in decidualized stromal cells of the uterine endometrium and in ovarian granulosa cells. IGFBP-1, together with IGFs, insulin, ovarian steroids, cytokines, and other factors, is involved in a complex system which regulates menstrual cycles, ovulation, decidualization, blastocyst implantation, and fetal growth. (ABSTRACT TRUNCATED) PMID- 9402141 TI - Lysophosphatidic acid and platelet-derived growth factor synergistically stimulate growth of cultured rat mesangial cells. AB - Lysophosphatidic acid (LPA) is a structurally simple, platelet-derived phospholipid, capable of eliciting a variety of physiological responses. We have demonstrated previously that LPA elicited a marked contractile response in rat mesangial cells (Inoue CN, Forster HG, Epstein M. Circ Res 77:888-896, 1995). In the present study, we examined the potential of this vasoactive substance to induce mesangial cell proliferation. Serum-starved quiescent rat mesangial cells were incubated with either LPA or in combination with platelet-derived growth factor (PDGF). DNA synthesis was assessed by [3H]thymidine incorporation after 24 hr, and cell numbers were determined at 0, 4, and 7 days. LPA- (1 nM-30 microM) stimulated mesangial cell DNA synthesis in a dose-dependent manner. The DNA synthesis stimulated by PDGF (1-100 ng/ml) was characterized by a bell-shaped response curve with a maximum at 40 ng/ml PDGF. The ability of LPA (30 microM) to synergize PDGF was observed over the entire range of PDGF concentrations (1-100 ng/ml). Under optimal concentrations of LPA/PDGF (30 microM40 ng/ml, respectively), mesangial cells displayed a 67-fold increase in [3H]thymidine incorporation, and a 1.9-fold (Day 4) and 2.5-fold (Day 7) increase in cell number as compared with that of quiescent mesangial cells. With an in vitro assay with myelin basic protein as the substrate, both LPA and PDGF induced stimulation of mitogen-activated protein (MAP) kinase activity. In addition, LPA augmented PDGF-induced increase in MAP kinase activity. In summary, these results demonstrate that LPA is mitogenic alone and also acts synergistically in combination with PDGF to promote mesangial cell proliferation. We postulate that these actions of LPA have the potential to play a crucial role in the mitogenic response of mesangial cells seen in a wide array of inflammatory and thrombotic glomerular disorders. PMID- 9402142 TI - Age-related changes in glucose transporter-one mRNA structure and function. AB - To determine the molecular mechanisms of age-related changes in the expression of glucose transporter 1 (GLUT-1) mRNA in cerebral tissue, male Fischer 344 rats at 4, 12, and 24 months of age were studied. The GLUT-1 mRNA in cerebral tissue was not significantly different among the various age groups. The in vitro translatability of GLUT-1 mRNA of 24-month-old rats (0.867 +/- 0.066 arbitrary units) was significantly lower than that in 4-month-old (1.403 +/- 0.153) P < 0.01 or 12-month-old rats (1.387 +/- 0.122) P < 0.01. The poly (A) tail length of GLUT-1 mRNA decreased from 200-350 nt in 4-month-old rats to only 50-100 nt in 24 month-old rats. Twelve-month-old rats also showed reduced poly (A) tail lengths. The poly (A) tail of G3PDH mRNA was not altered with age. The changes in GLUT-1 mRNA translatability did not correlate with GLUT-1 content in total cerebral tissue homogenate or in isolated cerebral microvessels, suggesting that GLUT-1 protein turnover is altered with age. It is concluded that aging is associated with significant changes in the structure and function of GLUT-1 mRNA. PMID- 9402143 TI - Cytokine dysregulation and increased oxidation is prevented by dehydroepiandrosterone in mice infected with murine leukemia retrovirus. AB - The effects of murine leukemia retrovirus infection on production of cytokines was investigated in mice fed different doses of dehydroepiandrosterone (DHEA). Young C57BL/6 female mice were injected with LP-BM5 murine retrovirus or were kept as uninfected controls. Two weeks later, each group was divided into subgroups: fed unsupplemented AIN 93 diet as the control, or diets supplemented with 0.02% DHEA (0.9 mg/mouse/day) or 0.06% DHEA (2.7 mg/mouse/day). The uninfected mice supplemented with 0.06% DHEA showed a significant (P < 0.05) increase in interleukin-2 (IL-2) and gamma-interferon (IFN-gamma) production, and hepatic vitamin E levels. Retroviral infection induced severe oxidative stress that was reduced by DHEAS supplementation in retrovirally infected mice. DHEA supplementation prevented the retrovirus-induced loss of cytokines (IL-2 and IFN gamma) secretion by mitogen stimulated spleen cells. DHEA also suppressed the production of cytokines interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF alpha) by T helper 2 (Th2) cells which were otherwise stimulated by retrovirus infection. Thus, immune dysfunction and increased oxidation induced by murine retrovirus infection were largely prevented by DHEA. PMID- 9402144 TI - Pirfenidone attenuates bleomycin-induced changes in pulmonary functions in hamsters. AB - The antifibrotic potential of pirfenidone (5-methyl-1-phenyl-2-[1H]-pyridone) was examined in a single intratracheal bleomycin dose hamster model of pulmonary fibrosis. Bleomycin-induced fibrosis and the effectiveness of pirfenidone treatment were assessed by measuring pulmonary functions (Cqst, TLC, VC, IC, FRC, RV) and the level of hydroxyproline in whole lung homogenates. Thirty-five male golden Syrian hamsters were randomized into four experimental groups: saline instilled and fed a control diet of rat chow (SCD, n = 8); saline instilled and fed the control diet containing 0.5% (w/w) pirfenidone (SPD, n = 8); bleomycin instilled and fed the control diet (BCD, n = 7); and bleomycin instilled and fed the control diet containing 0.5% pirfenidone (BPD, n = 10). Twenty-one days following bleomycin instillation Cqst/TLC, TLC, VC, and IC were significantly reduced and total lung hydroxyproline levels were significantly increased in the BCD and BPD groups as compared with the SCD and SPD groups, respectively. Pirfenidone ingestion significantly attenuated these bleomycin-induced pertubations in pulmonary functions and lung hydroxyproline levels (BCD versus BPD). The data obtained in this study provide evidence of the benefical effects of pirfenidone in the hamster model of bleomycin-induced pulmonary fibrosis both at the functional and biochemical level. PMID- 9402145 TI - Chronic underfeeding increases the positive feedback efficacy of estrogen on gonadotropin secretion. AB - Reduced caloric intake has been shown to inhibit reproductive cycles in females of several mammalian species. Previous studies have shown that increased negative feedback efficacy of estrogen on gonadotropin secretion may be responsible. The present study was designed to test the alternate hypothesis that caloric restriction alters the positive feedback efficacy of estrogen on gonadotropin secretion. Adult, cycling female rats were placed on reduced food intake (R) equal to 50% of that consumed by ad/libitum-fed controls (C). When R rats stopped cycling, both R and C rats were ovariectomized (OVX) and immediately implanted subcutaneously with a Silastic capsule containing 100 microg 17beta-estradiol (E2). Blood samples were obtained at 0900-1000 hr and 1600-1730 hr on Days 2, 4, 6, 8, 10, 12, and 14 after OVX and implantation. Follicle-stimulating hormone (FSH), luteinizing hormone (LH), and E2 were measured by radioimmunoassay in duplicate aliquots. Results indicate that underfed female rats retain the ability to respond to elevated estrogen levels with an afternoon surge of gonadotropin which is present for at least 14 days for LH. By contrast, FSH surges in R rats became progressively smaller and were no longer significant after Day 10. The present results also demonstrate that the response of R rats to elevated estrogen levels is significantly greater than that of C rats on Days 2-4 for FSH and 2-14 for LH. It is concluded that an inability to respond to elevated estrogen levels with an afternoon LH surge is not the cause of the cessation of normal estrous cycles. The progressive decrease in the afternoon surge of FSH may be, at least partly, responsible for the decreased follicular development observed in underfed rats. Possible explanations of the enhanced LH response to the positive feedback of estrogen are discussed. PMID- 9402146 TI - An innate natural antibody is reactive with a cryptic sequence of lactoferrin exposed on sperm head surface. AB - Lactoferrin (LF), an 80-kDa glycoprotein of ubiquitous occurrence in body fluids, is multifunctional and capable of assuming different configurations to serve those functions. The capacity of LF to undergo endocytosis and the recent demonstration of LF binding to sequence specific DNA indicate that a function or capability of LF, in addition to iron chelation, bacteriostasis, and receptor specific lymphocyte binding, may be that of gene activation or silencing. The data of this report present a human physiological system, that of sperm entry into the oocyte in performance of fertilization in which, since LF is a component of the sperm protein coat, that capability could be expressed. However, the configuration of LF in that locus is one in which a revealed cryptic sequence provides the specific binding site for a natural antibody present in the fertilization milieu. The presence of that antibody suggests that a system of control of the potential interaction of LF with the intra-ooplasmic DNA, that of gametes or pronuclei, is operative. The configuration of LF on the sperm surface and designation of the reactive site for the natural antibody were enabled by a monoclonal antibody secreted by a hybridoma derived from a human cord blood B cell. Thus, in addition to information concerning the molecular flexibility of LF, these observations support the proposition that the repertoire of natural antibodies provides an innate homeostatic system, with each antibody serving a specific role. PMID- 9402147 TI - Xanthine oxidase decreases production of gut wall nitric oxide. AB - Multiorgan failure is often the lethal outcome of intratracheal aspiration of acidic gastric juice. The pathogenesis of multiorgan failure may involve a systemic imbalance between pro-inflammatory and anti-inflammatory factors. In an anesthetized rat model, intratracheal instillation of HCl elicited intestinal inflammation which was exaggerated by xanthine oxidase (XO) and attenuated by nitric oxide (NO). We hypothesized that XO may mediate injury in part by suppression of NO formation. Therefore, we measured intestinal tissue concentrations of the stable NO oxidative metabolites (NO2- and NO3-) following intratracheal (IT) instillation of NaCl or HCl alone or in combination with interventions aimed at increasing or decreasing XO activity. Compared with IT NaCl (control treatment) jejunal tissue NO2- and NO2- + NO3- concentrations were increased by allopurinol pretreatment, which inhibits XO, and were decreased by systemically administered XO, as well as by IT HCl. The decreased NO2- and NO2- + NO3- concentrations found following IT HCl were completely reversed by either allopurinol or by systemically administered L-arginine (the precursor of NO). We conclude that manipulation of the pro-inflammatory XO system has a reciprocal effect on the intestinal anti-inflammatory NO system in either the undamaged or the endobronchially acidified lung model. PMID- 9402148 TI - The renal handling of IgG in the aging rat and in experimental kidney disease. AB - The urinary excretion of total protein, low-MW proteins, albumin, high-MW proteins, and intact IgG was measured in male Wistar rats between the ages of 5 52 weeks, and in rats with experimentally induced glomerular or tubular proteinuria. About 25% of aging rats spontaneously developed focal glomerulosclerosis and a mild glomerular proteinuria. By age 52 weeks, total protein excretion in rats with glomerulosclerosis exceeded that of unaffected rats by a factor of seven (39.5 vs 5.4 mg/24 hr x 100 g body wt), and albumin excretion was seven times higher than IgG excretion in affected rats (21.2 vs 2.9 mg/24 hr x 100 g body wt). Rats with chromate toxicity exhibited a reversible tubular proteinuria, with low-molecular weight protein excretion reaching 16.8 mg/24 hr x 100 g body wt (75% of total protein excretion) at the time of peak toxicity. IgG excretion remained less than 0.6 mg/24 hr x 100 g body wt. Aminonucleoside induced a massive but reversible glomerular proteinuria (204 mg/24 hr x 100 g body wt), with IgG excretion reaching 11.4 mg/24 hr x 100 g body wt (6% of total protein excretion) at the time of peak toxicity. Biochemical and immunochemical studies showed that, while some intact IgG is present in normal rat urine, most IgG immunoreactivity is derived from low-molecular weight catabolic fragments of IgG which interfere with the immunoassay of intact urinary IgG. One of these fragments, probably Fc fragment, may be involved in the pathogenesis of focal segmental glomerulosclerosis. PMID- 9402149 TI - In pursuit of drugs for American trypanosomiasis: evaluation of some "standards" in a mouse model. AB - Forty-nine "standard" compounds known to be useful in the treatment of other diseases were tested for their suppressive activity against the trypomastigotes of Trypanosoma cruzi-infected mice. The most active was the antidepressant protriptyline, which was almost three times as effective as the reference drug, nifurtimox. A major value of the present data is to demonstrate the refractoriness of the T. cruzi parasite against many of the drug standards that have known biological activity. PMID- 9402150 TI - Effect of Alzheimer's brain extracts on dynein immunoreactivity in PC12 cells. AB - The neurodegenerative process in Alzheimer's disease (AD) has been suggested to occur as a consequence of microtubule disruption and subsequent loss of intracellular transport. Structural microtubule-associated proteins (MAPs) have been investigated for their role in the etiology of AD, but dynein, a force producing MAP which mediates intracellular transport, has not been examined. In this report, dynein (MAP1C) immunoreactivity in AD brain tissue homogenates was observed increased 3.7-fold compared with control brain homogenate preparations. Similarly, NGF-differentiated PC12 cells cultured in the presence of soluble extracts prepared from AD brain tissue homogenates, exhibited an approximate 15 fold increase in dynein immunoreactivity compared to that of control brain tissue extracts. In contrast, AD clarified extracts had little effect upon "kinesin like" protein immunoreactivity increased (approximately 2-fold); whereas, tau immunoreactivity was observed to be moderately increased (5-fold) over that of control brain extract treated PC12 cells. Chemical dephosphorylation and alkaline phosphatase treatment of AD extract-treated PC12 cell lysate prior to Western blotting resulted in complete loss of immunoreactivity, suggesting the dynein being monitored is a phosphorylated isoform. Furthermore, treatment of clarified brain tissue extracts with trypsin and (NH4)2SO4 suggests the endogenous elements giving rise to increased PC12 cell dynein intermediate chain immunoreactivity to be proteinaceous in nature. The observed increase in dynein intermediate-chain dynein immunoreactivity following exposure of neuronal cells to endogenous elements of AD brain may be reflective of dynein-microtubular array differences. Such an approach may be useful in assessing the effect of endogenous biomolecules on retrograde axonal transport in neuronal culture models. PMID- 9402151 TI - Stimulation of intestinal growth is associated with increased insulin-like growth factor-binding protein 5 mRNA in the jejunal mucosa of insulin-like growth factor I-treated parenterally fed rats. AB - Surgically stressed rats maintained with total parenteral nutrition (TPN) exhibit jejunal atrophy, which can be attenuated by insulin-like growth factor-I (IGF-I) but not by growth hormone (GH) treatment. In order to understand the basis for the selective action of IGF-I, the levels of mRNAs encoding IGF-I, IGF-binding proteins (IGFBPs), IGF-I receptor, and GH receptor/binding protein (GHR/GHBP) were determined in rats given TPN and treated with GH, IGF-I, or GH + IGF-I. GH treatment significantly stimulated hepatic IGF-I mRNA. IGF-I treatment did not alter liver IGF-I mRNA, nor was there any evidence for interaction between GH and IGF-I. Jejunal mucosa IGF-I mRNA was extremely low and was not altered by TPN or by any of the hormonal treatments. The inability of GH to stimulate jejunal growth was not associated with a deficiency in GHR/GHBP mRNA. In jejunal mucosa, IGF-I and GH treatment independently and synergistically stimulated IGFBP-3 mRNA. IGF-I stimulated jejunal IGFBP-5 mRNA, but GH had no effect on IGFBP-5 mRNA. The levels of IGF-I receptor and IGFBP-1, 2, 4, and 6 mRNAs were extremely low and/or were not altered by any of the treatments. These results suggest that the ability of exogenous IGF-I, but not GH, to induce IGFBP-5 mRNA in jejunal mucosa may lead to the selective growth-promoting effect of IGF-I. Jejunal mucosa IGFBP-3 mRNA levels were not correlated with altered growth. We postulate that IGFBP-5 positively modulates the anabolic effects induced by exogenous IGF-I in the jejunum. PMID- 9402152 TI - Disruption of rat estrous cyclicity by the environmental estrogen 4-tert octylphenol. AB - 4-tert-Octylphenol (OP) is a prevalent environmental pollutant which binds to estrogen receptors and exerts estrogenic actions in vitro. The effects of OP in vivo on mammalian female reproduction are not known. We investigated whether (i) exposure of neonatal rats to OP interfered with the onset of vaginal opening or their ability to have regular estrous cycles as adults and (ii) exposure of adult rats to OP interfered with estrous cyclicity and ovulation. Injection of 1 mg OP in corn oil sc on the day after birth did not affect the day of vaginal opening. However, 9 of 11 OP-treated rats were in persistent vaginal estrus when examined at three months after birth compared with 0 of 9 corn oil-injected controls, which cycled regularly. Ten of eleven neonatal rats injected with 1.7 mg of the estrogenic pesticide methoxychlor also were in persistent estrus at 3 months after birth, and all 10 neonatal rats injected with 1 mg of 2,4,5 trichlorophenol, which is apparently nonestrogenic, cycled regularly. Injection of 20 or 40 mg OP in corn oil vehicle sc three times weekly into previously untreated adult cyclic rats caused persistent estrus in 2 of 6 and 16 of 21 rats, respectively. Injections were continued for three more weeks in 5 of the 16 rats rendered persistent estrus by the 40 mg OP treatment. These rats remained in persistent estrus for the additional 3-week period. The other 11 persistent estrous rats in the 40 mg treatment group started to cycle regularly within 5-7 days after the last injection. Unlike pentobarbital, injection of OP into cyclic rats during the afternoon of proestrus did not block ovulation. These results provide strong evidence that OP acts like estrogen in vivo in both neonatal and adult female rats to exert effects that block reproductive cyclicity. PMID- 9402153 TI - In vitro effects of boron-containing compounds upon glioblastoma cells. AB - Boron-neutron capture therapy (BNCT) is currently under investigation as a novel therapeutic modality for glioblastoma. This study was undertaken to determine whether boron-containing compounds 4-borono-2-fluoro-D,L-phenylalanine (FBPA) and FBPA-fructose have direct effects upon kinetics of A172, a glioblastoma cell line. Flow cytometry analyzed cell-cycle distribution and S-phase kinetics (bromo deoxyuridine [BUdR] incorporation). BUdR incorporation was increased during a 1 hr pulse after 24-hr or 72-hr exposure of cells to varying concentrations of FBPA or FBPA-fructose. Results suggest that boron-containing compounds may effect cell kinetics apart from neutron activation, and this effect should be further evaluated for potential impact upon tumor responsiveness to BNCT. PMID- 9402154 TI - Heme induces the expression of adhesion molecules ICAM-1, VCAM-1, and E selectin in vascular endothelial cells. AB - Heme is an important immunostimulating agent and oxidative factor contributing to endothelial cell activation. To investigate the mechanism of heme-induced endothelial cell activation, we analyzed the effect of heme and the inflammatory cytokine, tumor necrosis factor-alpha (TNF-alpha), on the expression of the heme degrading stress protein, heme oxygenase (HO), and adhesion molecules in human umbilical vein endothelial cells (HUVEC). Indirect immunofluorescence double labeling studies demonstrated a simultaneous increase of ICAM-1 and HO-1 after exposure of cells to heme for 24 hr. Co-expression of HO-1 and ICAM-1 was also demonstrated in TNF-alpha-exposed cells. Dot blot immunoassay and quantitative analysis by ELISA demonstrated that heme treatment for 24 hr caused a 2-fold increase in ICAM-1 expression (P < 0.002) compared with quiescent cells, while in cells stimulated by TNF-alpha for 24 hr ICAM-1 gene expression increased by 5 fold. Moreover, heme exposure also resulted in a marked increase in VCAM-1 and E selectin expression (three and four times over control levels, respectively). On the other hand, TNF-alpha treatment showed similar expression levels for VCAM-1 and E selectin, compared with stimulation by heme (100 microM). The level of HO activity in endothelial cells exposed to heme or TNF-alpha was increased from 24.7 +/- 5.7 pmol bilirubin/mg protein/min in control to 70.0 +/- 9.5 and 36.7 +/ 3.1 pmol bilirubin/mg protein/min in heme- and TNF-alpha-stimulated cells, respectively. These results suggest that upregulation of ICAM-1, VCAM-1, and E selectin expression is associated with oxidative stress induced by hemoglobin/heme and that HO-1 may play a modulating role via its ability to degrade heme to a substance with antioxidant properties. PMID- 9402156 TI - Situation of medical oncology in Hong Kong remains unchanged. PMID- 9402155 TI - Human embryonic cells raise some hopes, but mainly questions. PMID- 9402157 TI - Swiss heroin success prompts consideration of liberalization. PMID- 9402158 TI - Death rates in Russia rise dramatically: worse than under Stalin. PMID- 9402159 TI - To what extent are cancer clinical trials imperilled in the UK? PMID- 9402160 TI - Will Jospin solve Villejuif's problems? PMID- 9402161 TI - Eugenics: did Hitler merely emulate others? PMID- 9402162 TI - High-dose chemotherapy and autotransplants: a time for guidelines. PMID- 9402163 TI - Prognostic factors in salvage therapy of ovarian cancer. PMID- 9402164 TI - Anthracyclines-paclitaxel combinations in the treatment of breast cancer. PMID- 9402165 TI - Ethical issues in genetic screening for cancer. AB - Genetically-based diseases with a late onset, such as BRCA1-dependent breast cancer or Huntington's disease, can be predicted by the screening of relevant mutations in members of high-risk families. Genetic screening is characterized by a conflict between respect for autonomy--e.g., the 'right not to know'--and responsibility toward future generations (the 'duty to know' for the sake of one's descendants). Other ethical conflicts are related to uncertainty as to benefits deriving from screening for mutations, since for most conditions no clearly effective therapeutical strategy has as yet been defined. In addition to monogenic high-penetrance conditions, polygenic low-penetrance susceptibility is attracting increasing attention, in particular with respect to environmental genetic interactions (metabolic polymorphisms). A simple approach to genetic screening would be to weigh the benefits and costs of genetic screening against those of primary prevention, and a superficial conclusion might be that genetic screening is less expensive and, overall, more practicable than restriction of toxic exposures or other known risk factors for the disease. Economic advantage notwithstanding, however, giving precedence to screening over primary prevention would be unacceptable. A serious hazard of genetic screening is the implicit limitation of research efforts aimed at primary prevention, and a serious drawback is its potential application for selection of nonsusceptible employees. The principle of equity is easily violated by genetic screening of workers in view of the fact that genetically-based metabolic polymorphisms are distributed unevenly among different ethnic groups. PMID- 9402166 TI - Four-step high-dose sequential chemotherapy with hematopoietic progenitor-cell support as induction treatment for patients with solid tumors. AB - BACKGROUND: Despite recent progress in modern chemotherapy, metastatic solid tumors still have a poor outcome. The delivery of increased dose intensities of cytotoxic agents could improve response rates. We assessed the feasibility and safety of a high-dose sequential chemotherapy program in chemotherapy-naive patients with solid tumors. PATIENTS AND METHODS: Thirty patients (14 with carcinoma of unknown primary site, seven with metastatic breast cancer, six with small-cell lung cancer, and three with other diseases) were treated by an induction therapy regimen consisting of four cycles of high-dose chemotherapy with hematopoietic progenitor cell and growth factor support. Peripheral blood progenitor cells were collected by apheresis as the leukocyte counts recovered from the nadir induced by the first cycle of chemotherapy (doxorubicin 75 mg/m2, cyclophosphamide 6000 mg/m2). Patients then received two cycles of etoposide (800 mg/m2) and carboplatin (900 mg/m2) separated by one cycle of doxorubicin (75 mg/m2) and cyclophosphamide (3000 mg/m2). G-CSF (5 microg/kg/d) was given until engraftment. Cycles were scheduled to be delivered every three weeks. RESULTS: A total of 108 cycles of chemotherapy were administered. Six patients went off study before the end of the program (three because of progressive disease, three because of toxicity). After the first cycle, a median number of 10 x 10(6)/kg CD34+ cells (range 8-30) were collected. The median number of apheresis procedures was 1 (range 1-3). From cycle 2 to cycle 4, the median number of days when there was an absolute neutrophil count of less than 500/microl increased from three to five, and the median number of days when the platelet count was less than 25,000/microl increased from three to six. Episodes of febrile neutropenia occurred in 36%, 50% and 46% of cycles during cycles 2, 3 and 4, respectively. The median numbers of days between cycle 1 and cycle 2, cycle 2 and cycle 3, cycle 3 and cycle 4 were 24 (range 20-30), 22 (range 20-36) and 22 (range 18-35), respectively. There were no treatment-related deaths. Non hematologic toxicity included severe (WHO grades 3 or 4) nausea/vomiting in 19 (18%) cycles, mucositis in 8 (7%) cycles and diarrhea in 7 (6%) cycles. CONCLUSION: Support with hematopoietic progenitor cells and growth factors allows the timely administration of repetitive cycles of high-dose chemotherapy in chemotherapy-naive patients, resulting in a significant increase in dose intensity. Toxicity is noteworthy but manageable and does not compromise further therapy. PMID- 9402167 TI - Multiple cycles of high-dose doxorubicin and cyclophosphamide with G-CSF mobilized peripheral blood progenitor cell support in patients with metastatic breast cancer. AB - BACKGROUND: In a previous study we applied doxorubicin and cyclophosphamide in a dose-intensive regimen with GM-CSF to patients with metastatic breast cancer (MBC). That treatment failed to prolong the remission duration compared to conventional-dose chemotherapy. In the present study we escalated the dosages of the same agents to: 1) determine the maximum tolerated dosages (MTD) when given for three cycles with G-CSF mobilised peripheral blood progenitor cell (PBPC) reinfusion and 2) evaluate the antitumour effect of this regimen. PATIENTS AND METHODS: For mobilisation of PBPC, G-CSF 15 microg/kg/day was given subcutaneously (s.c.), and in subsequent cohorts leucapheresis was started on days 3, 4 or 6. The intention was to treat MBC patients with three cycles of doxorubicin and cyclophosphamide at a starting dose of doxorubicin 90 mg/m2 and cyclophosphamide 1000 mg/m2. Dosages were then escalated in subsequent cohorts of at least three patients. In case of dose-limiting mucositis, only the dose of cyclophosphamide was escalated in the next cohort. RESULTS: Twenty-one patients entered this protocol, of which 18 patients received high-dose chemotherapy. The mobilisation of PBPC using G-CSF only was sufficient for three cycles of high dose chemotherapy in 10 of 21 (47%) patients. Mucositis precluded dose escalation of doxorubicin beyond 110 mg/m2. The MTD in this combination was 110 mg/m2 for doxorubicin, and 4 g/m2 for cyclophosphamide, with haemorrhagic cystitis being the dose-limiting toxicity. The overall response rate was 78% (95% confidence interval (95% CI): 57%-97%), with 22% (95% CI: 3%-41%) complete responses. CONCLUSION: The MTD of this three cycle high-dose regimen was doxorubicin 110 mg/m2 and cyclophosphamide 4 g/m2 with mucositis and cystitis being dose-limiting toxicities. Although the primary aim was not the evaluation of antitumour effect, this high-dose regimen does not appear to provide an improvement of treatment results in comparison with our previous study with the same drugs at moderately high-dosages without stem cell support. PMID- 9402168 TI - Predictors of response to subsequent chemotherapy in platinum pretreated ovarian cancer: a multivariate analysis of 704 patients [seecomments]. AB - BACKGROUND: The probability of response to chemotherapy following platinum based treatment in ovarian cancer has usually been related to the 'platinum-free interval'. However, in a recent European-Canadian trial of paclitaxel, serous histology, tumor bulk, and hemoglobin, but not treatment free interval, were predictors of response. To determine if these observations were unique to this study (or this drug), data from other active agents given as second- or third line treatment in ovarian cancer were obtained and analyzed. METHODS: In the first part of the study, results of trials in 1185 platinum pretreated ovarian cancer patients were obtained on six agents: paclitaxel, epirubicin, docetaxel, carboplatin, irinotecan, and gemcitabine. Response results according to histology, baseline hemoglobin, tumor size and time from last chemotherapy were determined for each agent and the Cochran-Mantel-Haenszel procedure was used to obtain an overall assessment of significance for each factor. In the second part of the study, individual data from 704 patients in four studies (three agents: paclitaxel, docetaxel and epirubicin) were pooled for univariate and multivariate analysis of factors predictive of response. RESULTS: In the analysis of results of individual agents all factors examined were significant predictors of response: serous histology (P = 0.001), tumor size < or = 5 cm (P = 0.02), normal baseline hemoglobin (P = 0.003), and > or = 6 mo since last treatment (P = 0.001). While these results were interesting, they did not supply definitive answers regarding independent response predictors. Therefore a multifactor analysis was undertaken on the 704 patients for whom individual data were available. Of the 11 factors examined in a univariate analysis, 10 met the criteria for inclusion in a stepwise logistic regression. In the final model only 3 factors remained as significant independent predictors of response: serous histology (P = 0.009), no. disease sites (P = 0.003), and tumor size (P = 0.001). Time from last treatment, when evaluated as a continuous variable, was not in the final model and was highly correlated with tumor size (P = 0.0005). CONCLUSIONS: On the basis of this analysis, we conclude that tumor burden (as assessed by size of the largest lesion and number of disease sites) and histology are factors of importance in response to subsequent chemotherapy in relapsed ovarian cancer. Time from last treatment was correlated with tumor size in this data set and its effect on response was dependent on whether it was examined as a categorical or continuous variable, so we conclude it is not the sole critical factor of biologic importance. We recommend description of these factors in reports of phase II studies, confirmation of these findings in other data sets and further investigation of the mechanism of sensitivity of serous tumors. PMID- 9402169 TI - Cisplatin-, epirubicin- and paclitaxel-containing chemotherapy in uterine adenocarcinoma. AB - PURPOSE: To evaluate the toxic effects and antitumour activity of a multidrug regimen with cisplatin, epirubicin and paclitaxel (CEP) as initial therapy in patients with uterine adenocarcinoma. PATIENTS AND METHODS: Forty-nine patients with histologically-confirmed diagnoses of locally advanced, recurrent or metastatic cervical or endometrial adenocarcinoma entered the study. Treatment consisted of epirubicin (E) given at 70 mg/m2 followed by paclitaxel (P) (175 mg/m2 over three hours) and cisplatin (C) (50 mg/m2), repeated every three weeks. Eligibility criteria also included age < or = 75 years, no previous chemotherapy, no previous radiotherapy to the tumour parameters, bidimensionally-measurable lesions, no previous or ongoing cardiac disease, and renal and liver function within normal limits. Complete blood cell counts were repeated weekly, and tumor response was assessed every three cycles. A maximum of eight courses was administered in responding patients. RESULTS: From January 1996 to January 1997, 30 patients with endometrial adenocarcinoma and 19 with cervical adenocarcinoma entered the study, for a total of 213 cycles of treatment. In patients with endometrial carcinoma the overall clinical and pathological response rates were 73% (95% CI, range 54%-88%) and 35% (95% CI, range 16%-57%) respectively; in patients with locally advanced cervical carcinoma the overall clinical and pathological response rates were 64% and 62%. WHO grade 3-4 neutropenia occurred in 61% of the patients, with one possible toxic death. Retreatment had to be delayed for at least one week because of persisting neutropenia in 34% of the patients. Mild peripheral neuropathy and stomatitis were observed in 46% and 23% of the patients. One patient presented acute congestive heart failure after the third cycle of treatment. CONCLUSION: The high antitumour activity and the good tolerability of CEP suggest that this regimen should be prospectively compared to standard combinations as initial treatment of advanced endometrial carcinoma. PMID- 9402170 TI - Effect of age on the characteristics and clinical behavior of non-Hodgkin's lymphoma patients. The Non-Hodgkin's Lymphoma Classification Project. AB - BACKGROUND: The goals of this study are to describe the frequency, clinical characteristics, and outcome of the different non-Hodgkin's lymphomas according to age. PATIENTS AND METHODS: Patients included in the recently published analysis of the Non-Hodgkin's Lymphoma Classification Project were analyzed. All patients had their slides reviewed and classified by five independent expert hematopathologists. Lymphomas were classified according to the Revised European American Classification of lymphoid neoplasms. Sufficient data were available on 1283 cases. Five age groups were analyzed: < 35 years, 35-49 years, 50-59 years, 60-69 years, and > or = 70 years. RESULTS: Few differences were observed between the age groups with regard to lymphoma types and clinical characteristics. Anaplastic large cell lymphoma, Burkitt's lymphoma, and lymphoblastic lymphoma were observed more frequently in patients younger than 35 years, whereas small lymphocytic and lymphoplasmacytoid lymphomas were observed more frequently in patients older than 70 years. Mantle cell lymphoma and marginal zone lymphomas were observed more frequently in middle-aged patients. Poor performance status was more frequent in older patients, as was bone marrow infiltration, whereas spleen involvement was more frequent in younger patients. Young and older patients had a slightly worse age-adjusted International Prognostic Index score (P < 0.01). Complete response rates decreased with age from 68% in the youngest patients to 45% in the oldest patients (P < 0.0001). Median event-free survival and overall survival also decreased with age (P < 0.0001). CONCLUSIONS: Elderly patients have a poorer outcome than younger patients but age alone is not sufficient to discriminate patients with a poor outcome. However, the histologic type of lymphoma and clinical characteristics may define a subgroup of patients with a poor outcome in each age category. PMID- 9402171 TI - t(11;18)(q21;q21) is the most common translocation in MALT lymphomas. AB - BACKGROUND: Low grade malignant lymphomas arising from mucosa associated lymphoid tissue (MALT) represent a distinct clinicopathological entity. The cytogenetic findings and molecular genetics of MALT lymphomas remain minimally defined. Cytogenetic studies infrequently constitute part of the diagnostic work-up of MALT lymphomas, most commonly due to small biopsy size and their extranodal localization. Only 28 MALT cases with a clonal karyotype have been published to date. A number of chromosomal abnormalities have been observed with the majority of the cases featuring trisomy of chromosome 3 which is present in up to 78% of the cases. MATERIALS AND METHODS: A total of 116 cases of MALT lymphoma were diagnosed at BCCA between 1988 and 1997. Eleven cases of pathologically confirmed MALT lymphomas were subjected to cytogenetic analysis at the time of the initial evaluation. Eight of 11 cases yielded successful cultures and the presence of a clonal karyotype using standard cytogenetic methodology. In addition, a single case of orbital MALT lymphoma with a clonal karyotype has been obtained through our consultative practice from University of Nebraska Medical Center. These nine cases of MALT lymphoma with a clonal karyotype are the subject of this report. RESULTS AND CONCLUSION: In this study we report nine cytogenetically studied MALT lymphomas, three of which feature a novel t(11;18)(q21;q21) translocation which has also been observed in five other MALT cases described in the literature. This recurrent translocation is the most common translocation associated with MALT lymphomas being present in 33% (three of nine) of our cases and 18% (five of 28) of the previously published cases. The results suggest that a potentially important gene located at one of these breakpoints may be involved in the pathogenesis of MALT lymphomas. PMID- 9402172 TI - Intraclonal molecular heterogeneity suggests a hierarchy of pathogenetic events in Burkitt's lymphoma. AB - BACKGROUND: Burkitt's lymphoma is a B-cell neoplasm characterized by a chromosomal translocation involving the c-myc gene. BL may carry, besides the c myc translocation, several other lesions including a) mutations in c-myc, b) mutations in bcl-6, c) mutations in p53 and d) EBV genomes. In this report we describe a unique study of the timing of these genetic lesions during the evolution and progression of Burkitt's lymphoma. MATERIALS AND METHODS: From each of two patients with Burkitt's lymphoma, we established three different cell lines from different sites or at different times in the clinical course of the disease (diagnosis and relapse). Chromosomal aberrations were analyzed by karyotyping and the presence of molecular lesions determined by Southern blot, PCR, SSCP and sequence analyses. RESULTS: In each patient all the clones carry identical c-myc translocations, identical bcl-6 status (wild type or mutant) and the same productive VDJ rearrangement. However, within each individual patient, we could demonstrate the presence of intraclonal variation with respect to EBV, p53 mutations and c-myc mutations. CONCLUSIONS: c-myc translocation and bcl-6 mutations appear to be early events, mutations in the coding region of c-myc occur early but are an ongoing event, while mutations in the p53 gene seem to occur later. Discrete clonal bands reflecting independent EBV infection were observed in the cell lines from one HIV-associated Burkitt's lymphoma, suggesting the possibility that EBV infection may occur as a late event, at least in some HIV associated lymphomas. PMID- 9402173 TI - A prospective randomised trial of protracted venous infusion 5-fluorouracil with or without mitomycin C in advanced colorectal cancer. AB - BACKGROUND: To compare protracted venous infusion (PVI) 5-fluorouracil (5-FU) with and without mitomycin C (MMC) in a prospectively randomised study and analyse for tumour response, survival, toxicity and quality of life (QL). PATIENTS AND METHODS: Two hundred patients with advanced colorectal cancer received PVI 5-FU 300 mg/m2/day for a maximum of 24 weeks and were randomised to PVI 5-FU alone or PVI 5-FU + MMC 10 mg/m2 (7 mg/m2 from June 1995) 6 weekly for 4 courses. RESULTS: Overall response was 54% (95% confidence interval [CI] 44.1% 63.9%) with PVI 5-FU + MMC compared to 38% (95% CI: 28.3%-47.7%) for PVI 5-FU alone (P = 0.024). The median failure free survival was 7.9 months in PVI 5-FU plus MMC and 5.4 months with PVI 5-FU alone (P = 0.033) and at one year 31.9% for the combination compared to 17.7% for PVI 5-FU alone. Median survival was 14 months with MMC and 15 months in 5-FU alone; one-year survival 51.7% vs. 57.2%. PVI 5-FU + MMC caused more overall haematological toxicity but CTC grades 3/4 was increased only for thrombocytopaenia. Two patients treated with a cumulative dose of MMC of 40 mg/m2 developed haemolytic uraemic syndrome warranting the reduction in cumulative MMC dose to 28 mg/m2. The global QL scores were better for PVI 5-FU + MMC arm at 24 weeks, but the remaining QL data showed no differences. CONCLUSIONS: PVI 5-FU + MMC results in failure-free survival and response advantage, tolerable toxicity and better QL when compared to PVI 5-FU alone but no overall survival advantage. There is no irreversible toxicity with MMC at a cumulative dose of 28 mg/m2. PMID- 9402174 TI - Phase I study of daily times five topotecan and single injection of cisplatin in patients with previously untreated non-small-cell lung carcinoma. AB - BACKGROUND: The objectives were to determine the dose-limiting toxicity of topotecan in combination with cisplatin, to describe the principal toxicities, and to define the maximally-tolerated doses of the drugs in previously untreated patients with advanced non-small-cell lung carcinoma. PATIENTS AND METHODS: The study was designed to evaluate escalated doses of topotecan (starting at 0.75 mg/m2/day) as a 30-minute infusion daily for five consecutive days with a fixed clinically-relevant dose of 75 mg/m2 cisplatin given on day 1, every three weeks. RESULTS: Fifteen chemotherapy-naive patients entered the study and 14 were evaluable for toxicity. All 11 patients treated at the first topotecan/cisplatin dose level of 0.75/75 mg/m2, experienced at least one episode of grade 4 neutropenia. For six patients, absolute neutrophil counts were below 500/ml for more than five days, and two of them developed a grade 4 thrombocytopenia. At the next higher topotecan/cisplatin dose level (1.0/75 mg/m2), grade 4 neutropenia lasting longer than five days occurred in all three evaluable patients, including one patient who expired due to a severe neutropenia associated with sepsis. Non hematologic toxicities, predominantly nausea and vomiting, were mild to moderate in severity and manageable. Four patients had partial responses (30.7%; 95% confidence interval (9%-61%) of relatively short duration. CONCLUSION: Both severe neutropenia and thrombocytopenia precluded dose escalation of topotecan and cisplatin administered on this schedule. In previously untreated patients, the first topotecan/cisplatin dose level (0.75/75 mg/m2), was associated with intolerable myelosuppression, and, therefore, the dose levels evaluated in this study cannot be recommended for subsequent phase II investigations. The high toxicity of this schedule and the recent understanding of the pharmacokinetic interaction between those drugs may encourage the investigation of the alternate sequence of cisplatin after TPT in phase II studies. PMID- 9402175 TI - Long-term survivors of small-cell lung cancer (SCLC): a French multicenter study. Groupe d'Oncologie de Langue Francaise. AB - BACKGROUND: The aim of this study was to analyze SCLC patients beyond 30 months, particularly their outcome, their way of life, and factors which could influence relapses, second-primary cancers and death. PATIENTS AND METHODS: Between January 1986 and May 1995, 263 SCLC patients who survived longer than 30 months were included from 52 French institutions. The analysis was performed on the 155 cases confirmed by a pathologic review. RESULTS: Physical, mental and psychological states were considered as normal at 30 months in respectively 70.3%, 87.7% and 67.7% of patients, not influenced by prophylactic cranial irradiation, number of chemotherapy cycles, CCNU or cisplatin. Therapeutic sequelae were neurological impairment (13%), pulmonary fibrosis (18%) and cardiac disorders (11%) at 30 months. Return to work was possible for 40% of patients in the first two years following diagnosis. Among 43 relapsing patients, 33 benefited from a second-line treatment. Their median survival was 12 months since retreatment, and seven patients have survived again longer than 30 months. Age > 60 at the time of diagnosis was found as an independent factor increasing the risk of relapse beyond 30 months (OR = 2.46, IC 95% (1.16-5.26), P = 0.01). The risk of relapse became less than 10% beyond five years. Twenty patients (13%) developed a second primary cancer in a mean time of 58.6 months. The risk of second primary cancer was increased by a number of chemotherapy cycles > 6 (OR = 3.25, IC 95% (1.08 9.8) P = 0.02) and by an age > 60 (OR = 2.92, IC 95% (1.07-7.97), P = 0.03). Five and 10-year survival rates were respectively 68% and 44%. In these patients having reached a 30-month survival, three independent factors were predictive of a survival longer than five years: age < or = 60 at the time of diagnosis (OR = 2.85, IC 95% (1.23-6.6), P = 0.01), chest radiotherapy (OR = 3.1, IC 95% (1.28 7.69), P = 0.006) and absence of relapse (OR = 4.5, IC 95% (1.75-12.5), P = 0.002). This study suggests that: 1) therapeutic sequelae are rather mild, allowing return to work in 40% of patients; 2) relapsing 30-month survivors can benefit from second-line treatment; 3) SCLC cure can be achieved with a 10-year follow-up. PMID- 9402176 TI - Elevated ocular levels of vascular endothelial growth factor in patients with von Hippel-Lindau disease. AB - BACKGROUND: Von Hippel Lindau disease (VHL) is a rare autosomal dominant inherited disorder characterized by highly vascularized tumors in various organs. The abundant presence of endothelial cells in VHL tumors strongly suggest a role of the VHL tumor suppressor gene in the regulation of angiogenesis. Recently, in vitro studies have shown that the VHL tumor suppressor gene regulates the expression of vascular endothelial growth factor (VEGF). We investigated whether VHL patiens have increased levels of VEGF in their body fluids. PATIENTS AND METHODS: The concentration of VEGF was measured in fluid of the anterior chamber of the eye, serum, urine, and fluid from renal cysts of VHL patients and unaffected individuals by ELISA. In addition, levels of basic fibroblast growth factor (bFGF), interleukin-8 (IL-8) and endothelin-1 (ET-1) were measured in urine and serum of VHL patients and control subjects. RESULTS: In 80% of the VHL patients VEGF was detectable in aqueous fluid of the anterior chamber of their eyes. A strong positive correlation (r = 0.90) was found between the age of VHL patients and ocular VEGF concentrations. At comparable age, VEGF levels in ocular fluid of VHL patients were significantly higher (P < 0.001) than in unaffected subjects. No correlation was found between VEGF concentration and the presence of retinal angiomas. A 10 and 16 fold increase of VEGF concentration was seen in fluid from two independent VHL-related cysts as compared with VEGF serum levels of the same patient. The mean concentration of VEGF in serum of VHL patients (n = 15) (319 +/- 84 pg/ml) was higher than in matched controls (238 +/- 68 pg/ml; P = NS). The mean concentration of VEGF in urine of VHL patients (128 +/- 36 pg/ml) was lower than in matched controls (183 +/- 25 pg/ml; P = NS). Concentrations of VEGF did not correlate with the presence of VHL-related tumors. No differences were observed between concentrations of bFGF, IL-8 and ET-1 in serum and urine of VHL patients and matched controls. CONCLUSIONS: These findings support a role for the VHL tumor suppressor gene in the in vivo regulation of VEGF. PMID- 9402177 TI - Growth factor profiles of human gliomas. Do non-tumour cells contribute to tumour growth in glioma? AB - BACKGROUND: Growth factors play a role in proliferation and motility of malignant glial cells, through autocrine and paracrine mechanisms. Also, proliferation of non-tumour cells, e.g., endothelial cells, is likely to be controlled by growth factors. Several growth factors with their appropriate receptors can be involved, but studies on tissue specimens evaluating this in glioma are rare. MATERIALS AND METHODS: We evaluated the potential role of Transforming growth factor-alpha (TGF alpha) and Epidermal growth factor receptor (EGF-R), the Platelet-derived growth factor A- and B-chain (PDGF-A and PDGF-B) and its receptors (PDGFR alpha and PDGFR beta, and basic fibroblast growth factor (bFGF) in gliomas by analysing 86 of these tumours on the single cell level for the presence of immunoreactive growth factors and receptors. In a few cases double-staining experiments were done to directly visualize co-expression of factor and receptor. RESULTS: Multiple growth factors and their receptors are present in astrocytic tumours; the higher the grade, the more growth factors and the more positive cells are found. Oligodendroglial tumours and pilocytic astrocytomas showed little expression. Autocrine and paracrine mechanisms were frequently possible in the astrocytic tumours, often more than one loop could be involved. Interestingly, it was also frequently possible that non-tumour cells produced a growth factor for which the tumour cells expressed the receptor. CONCLUSIONS: Multiple growth factors appear to be involved in astrocytic tumours, with frequent autocrine and paracrine loops. Expression of these molecules seems to increase with increasing grade. The results argue for a contribution of non-tumour cells to the growth of a tumour. PMID- 9402178 TI - Complete spontaneous remission in a patient with metastatic non-small-cell lung cancer. Case report, review of the literature, and discussion of possible biological pathways involved. AB - Spontaneous remission of cancer (SR) is defined as a complete or partial, temporary or permanent disappearance of all or at least some relevant parameters of a soundly diagnosed malignant disease without any medical treatment or with treatment that is considered inadequate to produce the resulting regression. We report the case of a 61-year-old man who presented with extensive metatastic disease five months after pneumonectomy for poorly differentiated large cell and polymorphic lung cancer. A vast metastatic tumour mass of the abdominal wall was confirmed histolologically and there was clinical and radiographic evidence of liver and lung metastases. Eight months later, the patient was operated on for a hernia, which had developed in the inguinal biopsy scar and the surgeon confirmed complete clinical SR of the abdominal wall metastases. Again five months later there was no longer any radiologic evidence of liver and lung metastases. Complete remission has persisted more than five years. Histology of the primary and of the abdominal metastases were reviewed by several independent pathologists. SR is an extremly rare event in lung cancer. This is the first documented case of clinically evident visceral metastases of a bronchiogenic adenocarcinoma developing after complete resection of the primary and then showing complete SR. The epidemiology of SR is reviewed and possible mechanisms involved in SR are discussed. PMID- 9402179 TI - High-dose treatment with lanreotide of patients with advanced neuroendocrine gastrointestinal tumors: clinical and biological effects. AB - BACKGROUND: Neuroendocrine tumors usually present with inoperable metastatic disease and severe hormonal symptoms. Specific chemotherapy, alpha-interferon and the somatostatin analog octreotide are established therapies in these patients but all of them eventually fail. Other somatostatin analogs, e.g., RC-160 and lanreotide, are currently being studied in different doses and modes of administration. PATIENTS AND METHODS: Nineteen patients with advanced neuroendocrine gastrointestinal tumors [13 carcinoids and six endocrine pancreatic tumors (EPT)], liver metastases being present in 18, most of them heavily pretreated, were included. Seventeen out of 18 patients had somatostatin receptors demonstrated by octreotide scintigraphy. Lanreotide was given as four daily subcutaneous injections, starting with 750 microg/d, then increasing every week up to 12,000 microg/d after six weeks, a dose which was maintained, if tolerated, for 12 months, or until progression. RESULTS: There was a significant tumor size response (>50%) in one patient (5%), whereas 12 patients (70%) had tumor stabilization for 12 months. Bichemical tumor markers were significantly reduced at six months (urinary 5-hydroxyindoleacetic acid and plasma chromogranin) and 12 months (chromogranin) and the overall biochemical response rate was 58% with this high dose of lanreotide. Adverse events were observed and four patients stopped the treatment due to adverse events. Studies of tumor biopsies before and during treatment indicated induction of apoptosis in patients with tumor stabilization and biochemical response. CONCLUSION: High-dose treatment with lanreotide (12,000 microg/d) produced tumor size response in 5%, stabilization in 70% and a biochemical response in 58% of patients. These results should be related to the advanced stage of the disease as indicated by the mean duration of disease of more than four years, but they do not appear to be better than those achieved with standard doses of somatostatin analogs. However, in responding patients we observed induction of apoptosis in the tumors, a phenomenon not seen with regular doses of somatostatin analogs, but often produced by chemotherapeutic agents. PMID- 9402181 TI - Severe CPT-11 toxicity in patients with Gilbert's syndrome: two case reports. AB - BACKGROUND: CPT-11 is hydrolyzed to its active metabolite SN-38, which is mainly eliminated through conjugation by hepatic uridine diphosphate glucuronosyl transferases (UGTs) to the glucuronide (SN-38G) derivative. Preclinical studies showed that UGT*1.1 is the isozyme responsible for SN-38 glucuronidation. Patients with Gilbert's syndrome have deficient UGT*1.1 activity, therefore may have an increased risk for related CPT-11 toxicity. PATIENTS AND METHODS: Two patients with metastatic colon cancer and Gilbert's syndrome were treated with CPT-11 based chemotherapy. CPT-11, SN-38 and SN-38G pharmacokinetics parameters were obtained. Serum bilirubin was analysed by alkaline methanolysis and HPLC. RESULTS: Both patients presented grade 4 neutropenia and/or diarrhea (NCI-CTC) in every treatment cycle. Biliary index (after Gupta et al) values were well above 4000. CONCLUSION: We present the first clinical evidence linking bilirubin glucuronidation status and CPT-11 related toxicity. The severe toxicity experienced by the two patients with Gilbert's syndrome treated with CPT-11 based chemotherapy has a genetic basis. Individuals with Gilbert's syndrome have an enhanced risk for CPT-11 toxicity. Unconjugated serum bilirubin could be predictive parameter of CPT-11 toxicity. PMID- 9402182 TI - Ophthalmic toxicity following paclitaxel infusion. PMID- 9402180 TI - Cisplatin, gemcitabine and vinorelbine in locally advanced or metastatic non small-cell lung cancer: a phase I study. AB - PURPOSE: The objective of this study was to determine the maximally tolerable doses (MTDs) of vinorelbine (VNR) and gemcitabine (GEM) when combined with a fixed dose of cisplatin (CDDP). PATIENTS AND METHODS: Chemotherapy-naive patients with stage IIIB-IV non-small-cell lung cancer (NSCLC) received a fixed dose of CDDP (50 mg/m2) and escalating doses of VNR (starting from 20 mg/m2) and GEM (starting from 800 mg/m2) on days 1 and 8, every three weeks. The single escalation of GEM alone, by 200 mg/m2 at each step, was initially planned up to a dose of 1,200 mg/m2, to be followed by increments of the VNR dose of 5 mg/m2 at each step. RESULTS: Thirty-one patients were enrolled at five different dose levels. The escalation was stopped at level 4 (GEM 1,200 mg/m2 and VNR 25 mg/m2) since two of six patients of this cohort showed dose-limiting neutropenia at treatment cycle 1. Two different dose levels, GEM 1,200 mg/m2 + VNR 20 mg/m2, and GEM 1,000 mg/m2 + VNR 25 mg/m2 were fairly well tolerated. No treatment-related deaths occurred. Neutropenia was the main toxic effect, occurring in 76% of the total of 116 cycles delivered, and in 24% of them was of grades 3 or 4. A total of eight patients (26%) experienced grade 4 neutropenia lasting more than seven days; in five of them it occurred in the first course. Neutropenic fever was observed in four cases. Grade 4 thrombocytopenia occurred in only two patients. Non-hematologic toxicity was a minor problem in all patients but was never dose limiting. No complete responses were obtained, but sixteen out of 31 (52%) patients achieved partial responses. The median duration of response was 20 (range 6-56+) weeks, while at a nine-month median follow-up, the median survival time has not yet been reached. To date, 18 patients are still alive. The one-year projected survival for all patients was 51%. CONCLUSIONS: Our results show that CDDP, VNR and GEM can be safely given together without substantial reductions in their individual dose intensities. In our opinion, the dose level of GEM 1,000 mg/m2 + VNR 25 mg/m2 given in combination with CDDP 50 mg/m2 on days 1 and 8 of a three-week cycle can be recommended for phase II trials, since it provides a better balance in dose intensity of GEM and VNR. A phase II randomised study is underway to establish the activity of this new regimen (at the above-cited dose level) in chemo-naive NSCLC patients. PMID- 9402183 TI - Sublingual nitrate provides cause for fear of food in a carcinoid patient. PMID- 9402184 TI - T1rho-relaxation in articular cartilage: effects of enzymatic degradation. AB - Spin-lattice relaxation in the rotating frame (T1rho) dispersion spectroscopy and imaging were used to study normal and enzymatically degraded bovine articular cartilage. Normal specimens demonstrate significant T1rho "dispersion" (approximately 60 to approximately 130 ms) in the 100 Hz to 9 kHz frequency range. Proteoglycan-degraded specimens have 33% greater T1rho values than collagen-degraded or normal samples. T1rho-weighted images reveal structure not found in conventional T1- or T2-weighted images. Our results suggest that T1rho measurements are selectively sensitive to proteoglycan content. The potential of this method in distinguishing the early degenerative changes in cartilage associated with osteoarthritis is discussed. PMID- 9402185 TI - A multiple echo pulse sequence for diffusion tensor imaging and its application in excised rat spinal cords. AB - A new imaging sequence for rapid determination of the apparent self-diffusion tensor of water was developed and tested on fixed excised rat spinal cords. To reduce the time required to determine the tensor, the sequence utilized a new single-shot approach with multiple spin echoes. An assumption of cylindrical symmetry in the sample was made, thus requiring the measurement of only four of the six unique elements of the tensor. This assumption was found experimentally to be valid, and the results obtained using the new sequence were found to be quantitatively the same as results obtained using a standard spin-echo sequence. PMID- 9402186 TI - Functional MRI BOLD signal coincides with electrical activity in the rat whisker barrels. AB - Functional MRI (fMRI) provides a noninvasive method for mapping brain functional activity based on blood oxygenation level dependent (BOLD) image contrast that is primarily due to localized increases in perfusion. Recently, Malonek and Grinvald (Science 272:551-554, 1996) suggested that during sustained functional activation, the increases in perfusion were spread over a much larger area than the localized electrical activity. In this study, it is demonstrated that the spatial distribution of the BOLD fMRI signal during sustained stimulation of rat whiskers has the same spatial pattern and dimension as that of neuronal electrical activity in the rat whisker barrels. PMID- 9402187 TI - Energetics of 3.5 s neural activation in humans: a 31P MR spectroscopy study. AB - No direct information on brain energetics and energy-related compounds in the first seconds of physiological activation has been reported to date. In this study visual cortex high energy phosphate changes were monitored in 11 normal subjects during 3.5 s activation and the following 23.5 s by a simple 31P magnetic resonance spectroscopic method. An intraactivation decrease of phosphocreatine (PCr) was observed in all subjects, with changes in pH in three, one of them also presenting a change in adenosine triphosphate (ATP). In the subgroup of eight subjects without changes in pH, the mean rate of mean PCr decrease (D(PCr)) was 7.24 +/- 0.78%/s, and the postactivation mean rate of mean PCr recovery was <1/2 D(PCr). Short phasic neural activity requires a large amount of energy, i.e., at least three times basal consumption, in agreement with theoretical calculations. Additional energy demands in the visual cortex are several times those measured by positron emission tomography during prolonged stimulation studies, implying that mean energy requirements decrease with increases in duration of stimulation. During short activation, the vascular responses as detected by brain-mapping techniques (BMT) are preceded by an important reduction of the intracellular high-energy phosphate content, which returns to resting values during an interval that corresponds to the poststimulation return of BMT signals to baseline. PMID- 9402188 TI - Two-point Dixon technique for water-fat signal decomposition with B0 inhomogeneity correction. AB - To separate water and lipid resonance signals by phase-sensitive MRI, a two-point Dixon (2PD) reconstruction is presented in which phase-unwrapping is used to obtain an inhomogeneity map based on only in-phase and out-of-phase image data. Two relaxation-weighted images, a "water image" and a "fat image," representing a two-resonance peak model of proton density, are output. The method is designed for T1- or density-weighted spin-echo imaging; a double-echo scheme is more appropriate for T2-weighted spin-echo imaging. The technique is more time efficient for clinical fat-water imaging than 3PD schemes, while still correcting for field inhomogeneity. PMID- 9402189 TI - Simultaneous T2* and diffusion measurements with 3He. AB - It has recently been demonstrated that magnetic resonance (MR) imaging of human lungs and airways is possible with hyperpolarized gases such as 3He. Because the influence of the apparent transversal relaxation (T2* decay) and diffusion in 3He imaging have not been quantified, an imaging pulse sequence was developed to measure these two parameters simultaneously. The imaging pulse sequence generates two series of multiply recalled gradient echo images with both different echo spacings and diffusion-sensitizing gradients. From differences in exponential signal decay between the two series, T2* and diffusion coefficients, D, of both hyperpolarized and unpolarized 3He samples could be measured on a standard clinical imager using a home-built Helmholtz coil. In a hyperpolarized sample of pure 3He values of D = (1.8 +/- 0.2) x 10(-4) m2/s and T2* = 36 +/- 13 ms were measured, while D = (0.3 +/- 0.1) x 10(-4) m2/s and T2* = 136 +/- 66 ms were found in a Boltzmann-polarized 3He/O2 mixture. PMID- 9402190 TI - Dynamic 13C NMR analysis of pyruvate and lactate oxidation in the in vivo canine myocardium: evidence of reduced utilization with increased work. AB - In this work, substrate selection was monitored in the left ventricle of the canine myocardium by following pyruvate and lactate oxidation under in vivo conditions at basal and elevated workloads. These studies were conducted in the open chest model using dynamic 13C NMR techniques in the presence and absence of dichloroacetic acid (DCA), a well-known activator of pyruvate dehydrogenase (PDH). Following the infusion of (3-(13)C) pyruvate or (3-(13)C) lactate into the left anterior descending artery, highly variable 13C enrichments of glutamate, alanine, aspartate, and citrate were noted under low (RPP < 14,500 mmHg/min), intermediate (RPP = 15,000-25,000 mmHg/min), and high (RPP > 25,500 mmHg/min) rate pressure products (RPP). At low workloads, the myocardium typically oxidized the infused (3-(13)C) pyruvate or (3-(13)C) lactate and incorporated the labeled carbon into the glutamate pool as expected. However, in a few notable instances (n = 3), 13C-enriched pyruvate and lactate were unable to label the glutamate pool under in vivo conditions even at the lowest RPPs, indicating a lack of selection for these substrates by the tricarboxylic acid (TCA) cycle. Nonetheless, the levels of glutamate C4 enrichment observed at low workloads could usually be enhanced by infusion of DCA. Importantly, 13C NMR extract analysis revealed that (3-(13)C) pyruvate or (3-(13)C) lactate labeling of the glutamate pool was reduced (< 20%) at high workloads in spite of increased DCA concentrations. PMID- 9402191 TI - 19F NMR monitoring of in vivo tumor metabolism after biochemical modulation of 5 fluorouracil by the uridine phosphorylase inhibitor 5-benzylacyclouridine. AB - A uridine phosphorylase inhibitor, 5-benzylacyclouridine (BAU), has been utilized as biochemical modulator of 5-fluorouracil (5-FU) anti-tumor activity in a murine tumor model. The effect of BAU on 5-FU metabolism has been evaluated using in vitro and in vivo 19F NMR spectroscopy. The analysis of the NMR data revealed an increased formation and retention of fluorouracil nucleotides and fluorouridine in colon 38 tumors treated with the regimen containing BAU and a reduction in 5 FU catabolites (alpha-fluoro-beta-ureidopropionic acid and alpha-fluoro-beta alanine). In the normal tissues evaluated, the presence of BAU did not significantly alter the metabolism and presence of fluoropyrimidine species, indicating a more selective effect on tumor tissues. Therapy experiments on C57/BL6 mice bearing colon 38 tumor showed that the administration of 120 mg/kg BAU 30 min before 5-FU at 85 mg/kg, on a weekly basis, resulted in an increased antineoplastic effect compared to the same dose of 5-FU alone. A smaller dose of 5-FU (60 mg/kg) also administered 30 min after 120 mg/kg BAU caused a reduction in tumor growth similar to 5-FU alone. The addition of BAU to 5-FU (85 mg/kg) resulted in a slight increase, although statistically nonsignificant, in host toxicity without causing any toxic death during the chemotherapeutic treatment. 19F NMR spectroscopy is here shown to be a powerful technique to evaluate changes in the metabolism of fluoropyrimidines after the use of biochemical modulator and to allow a correlation between improved therapeutic response with the biochemical effects generated in tissues. PMID- 9402192 TI - The effect of perfusion on T1 after slice-selective spin inversion in the isolated cardioplegic rat heart: measurement of a lower bound of intracapillary extravascular water proton exchange rate. AB - Many NMR measurements of cardiac microcirculation (perfusion, intramyocardial blood volume) depend on some kind of assumption of intracapillary-extravascular water exchange rate, e.g., fast exchange. The magnitude of this water exchange rate, however, is still unknown. The intention of this study was to determine a lower limit for this exchange rate by investigating the effect of perfusion on relaxation time. Studies were performed in the isolated perfused cardioplegic rat heart. After slice-selective inversion, the spin lattice relaxation rate of myocardium within the slice was studied as a function of perfusion and compared with a mathematical model which predicts relaxation rate as a function of perfusion and intracapillary-extravascular exchange rate. A linear relationship was found between relaxation rate T(-1) and perfusion P normalized by perfusate/tissue partition coefficient of water, lambda: deltaT(-1) = m x deltaP/lambda with 0.82 < or = m < or = 1.06. Insertion of experimental data in the model revealed that a lower bound of the exchange rate from intra- to extravascular space is 6.6 s(-1) (4.5 s(-1), P < 0.05), i.e., the intracapillary lifetime of a water molecule is less than 150 ms (222 ms, P < 0.05). Based on this finding, the T1 mapping after slice-selective inversion could become a valuable noncontrast NMR method to measure variations of perfusion. PMID- 9402193 TI - Homocarnosine and the measurement of neuronal pH in patients with epilepsy. AB - Homocarnosine is a dipeptide of gamma-aminobutyric acid (GABA) and histidine found uniquely in the brain, most likely in a subclass of GABAergic neurons. By comparison of spectra from the occipital lobe of patients receiving a homocarnosine elevation drug to normal subjects we have assigned two elevated resonances in the short TE 1H MRS spectrum to homocarnosine. These resonances are partially resolved at 7.05 and 8.02 ppm in a short TE spectrum at 2.1 T when macromolecule resonances are removed by subtraction of a spectrum in which the metabolite resonances are nulled by inversion recovery. The chemical shift of both of these resonances is sensitive to pHi. By comparison with a titration curve the pHi was calculated from the downfield resonance to be 7.06 in the patient group which is similar to values reported using the P(i) resonance. Based on the in vivo results and theoretical considerations the potential sensitivity for using nonelevated homocarnosine to measure pH is similar to that of P(i) under physiological conditions. PMID- 9402194 TI - Localized q-space imaging of the mouse brain. AB - Localized q-space imaging was used to obtain water displacement profiles from mouse brain. These profiles take the form of unidirectional diffusive displacement probability distributions. Two groups of mice were studied, a normal group and a group in which surgery had been performed to produce a unilateral reduction in the supply of blood to the forebrain. q-Space measurements were made both in vivo and postmortem. The displacement profiles were characterized using the summary parameter prob[d < 10], which is the proportion of water molecules that undergo a net diffusive displacement that is less than +/-10 microm, during the diffusion period (50 ms). The range of prob[d < 10] values in the normal group was 0.71 to 0.77 in vivo compared with 0.78 to 0.87 in the impaired hemisphere of the surgically treated group. An increase in prob[d < 10] occurred postmortem to yield values in the range 0.79 to 0.81 and 0.80 to 0.89 in the normal and surgically treated group, respectively. These observations are consistent with the diffusion-weighted image intensity changes that occur after a period of ischemia. PMID- 9402195 TI - MR microimaging of the lung using volume projection encoding. AB - Radial acquisition (RA) techniques have been extended to produce isotropic, three dimensional images of lung in live laboratory animals at spatial resolution down to 0.013 mm3 with a signal-to-noise ratio of 30:1. The pulse sequence and reconstruction algorithm have been adapted to allow acquisition of image matrices of up to 256(3) in less than 15 min. Scan-synchronous ventilation has been incorporated to limit breathing motion artifacts. The imaging sequence permits randomizing and/or discarding selected views to minimize the consequences of breathing motion. The signal in lung parenchyma was measured as a function of flip angle (alpha) for different repetition times and found to follow the predictions for which there is an optimum excitation (Ernst) angle. A single T1 relaxation value of 780 +/- 54 ms fits all data from six guinea pigs at 2.0 T. This T1 value parameterizes the signal and allows for a priori optimization, such as calculation of the Ernst angle appropriate for lung imaging. PMID- 9402196 TI - Recording of EEG during fMRI experiments: patient safety. AB - The acquisition of electroencephalograms (EEG) during functional magnetic resonance imaging (fMRI) experiments raises important practical issues of patient safety. The presence of electrical wires connected to the patient in rapidly changing magnetic fields results in currents flowing through the patient due to induced electromotive forces (EMF), by three possible mechanisms: fixed loop in rapidly changing gradient fields; fixed loop in a RF electromagnetic field; moving loop in the static magnetic field. RF-induced EMFs were identified as the most important potential hazard. We calculated the minimum value of current limiting resistance to be fitted in each EEG electrode lead for a representative worst case loop, and measured RF magnetic field intensity and heating in a specific type of current-limiting resistors. The results show that electrode resistance should be > or = 13 k(omega) for our setup. The methodology presented is general and can be useful for other centers. PMID- 9402197 TI - On the SAR and field inhomogeneity of birdcage coils loaded with the human head. AB - Birdcage coils are widely used as a radiofrequency (RF) resonator in magnetic resonance imaging (MRI) because of their capability to produce a highly homogeneous B1 field over a large volume within the coil. When they are employed for high-frequency MRI, the interaction between the electromagnetic field and the object to be imaged deteriorates the B1-field homogeneity and increases the specific absorption rate (SAR) in the object. To investigate this problem, a finite-element method (FEM) is developed to analyze the SAR and the B1 field in a two-dimensional (2D) model of a birdcage coil loaded with a 2D model of a human head. The electric field, magnetic field, and SAR distributions are shown, and a comprehensive study is carried out for both linear and quadrature birdcage coils at 64, 128, 171, and 256 MHz. It is shown that to generate the same value of the B1 field, the SAR is increased significantly with the frequency, and for the same imaging method the SAR produced by a quadrature coil is significantly lower than that of a linear coil. It is also shown that the B1-field inhomogeneity is increased significantly with the frequency. PMID- 9402198 TI - Non-Fourier encoding with multiple spin echoes. AB - The advantages and limitations of multiple spin-echo sequences for non-Fourier encoding are investigated. Complications caused by improper encoding of alternate magnetization pathways due to imperfect refocusing pulses are analyzed. It is shown that mirror image ghosts result if the encoding RF pulse matrix is real valued. These ghosts can be avoided as long as the rows of the RF pulse matrix are conjugate symmetric, which implies that spatial profiles are real valued. Non Fourier encoding using bases derived from wavelet, Hadamard, and other real valued orthogonal functions does not result in a mirror ghost artifact. A RARE sequence for non-Fourier encoding has been implemented on a clinical imaging system and successfully applied for brain imaging. PMID- 9402199 TI - Magnetization transfer imaging in vivo of the rat brain at 4.7 T: interpretation using a binary spin-bath model with a superLorentzian lineshape. AB - Proton magnetization transfer contrast (MTC) imaging, using continuous wave off resonance irradiation, was performed on the rat brain in vivo at 4.7 Tesla. The observed MTC was studied in three different brain regions: the corpus callosum, the basal ganglia, and the temporal lobe. By systematically varying the offset frequency and the amplitude of the RF irradiation, the observed signal intensities for each region of interest were modeled using a system including free water and a pool of protons with restricted motions (R. M. Henkelman, X. Huang, Q. Xiang, G. J. Stanisz, SD Swanson, M. J. Bronskill, Magn. Res. Med. 29, 759 (1993)). Most of the relaxation parameters of both proton pools remained fairly constant for the three regions of interest, with a T2 value of about 9 micros for the immobilized protons, whereas the rate of exchange increased significantly from the temporal lobe to the corpus callosum. The optimal acquisition parameters for the improved MTC under steady-state saturation were found to be 2-10 kHz offset frequency and 500-800 Hz RF irradiation amplitude. Conversely, an irradiation amplitude of 3 kHz at an offset frequency of 12 kHz is required to minimize the direct effect of off-resonance irradiation. Such an approach could be extended to human brain imaging with the aim of characterizing tissue-specific disease. PMID- 9402200 TI - MRI quantitative myocardial perfusion with compartmental analysis: a rest and stress study. AB - K1 (first-order transfer constant from arterial plasma to myocardium for Gd-DTPA) and Vd (distribution volume of Gd-DTPA in myocardium) were measured in vivo in a canine model (n = 5) using MRI-derived myocardial perfusion curves and a compartmental model. Perfusion curves were obtained after a bolus injection of Gd DTPA (0.04 mM/kg) with an inversion-prepared fast gradient echo sequence. Myocardium and blood signal intensity were converted to a concentration of Gd DTPA, according to a model appropriate for short (<1 s) interimage intervals characteristic of cardiac-triggered acquisitions. Before dipyridamole-induced stress, K1 and Vd, obtained from the fit of the MRI-derived perfusion curves, were 6.2 +/- 1.4 (mHz) and 17.5 +/- 4.2%, respectively. After dipyridamole infusion, a K1 increase of a factor of 2.82 +/- 0.72 was measured (P = 0.003). No change was observed in Vd (P = 0.98). These results suggest that the K1 increase after dipyridamole reflects a flow-related effect that can be useful to quantify the MRI-derived perfusion curves. PMID- 9402201 TI - Contrast-agent phase effects: an experimental system for analysis of susceptibility, concentration, and bolus input function kinetics. AB - A system is presented for experimental arterial input function (AIF) simulation and for accurate measurement of the concentration, susceptibility effects, and magnetic moment of paramagnetic MR contrast agents. Signal effects of contrast agents are evaluated with a stable, well-characterized, and precise experimental setup. A cylindrical phantom and a closed-loop circulating flow system were designed for AIF simulation, assessment of the physical determinants of contrast agent phase effects, and measurement of contrast-agent properties under controlled conditions. A mathematical model of the AIF dynamics is proposed. From the experimental phase shift (delta phi), either the concentration or molar susceptibility, chiM, is determined. The linear dependence of delta phi on concentration and echo time (TE), the orientation dependence, and the lack of dependence on T1, T2, and diffusion time are proven precisely for water solutions under a wide variety of conditions. The measured effective magnetic moment of Gd+3, mu(eff), was 7.924 +/- 0.015 Bohr magnetons in agreement with the theoretical value of 7.937. PMID- 9402202 TI - Phase alignment of multiple surface coil data for reduced bandwidth and reconstruction requirements. AB - Multiple element surface coils are often used in clinical MRI to increase the image signal-to-noise ratio (S/N). Use of multicoils typically requires increased net sampling bandwidth and data processing for each coil element. A phase alignment technique is described which combines the signals from all coil elements before image reconstruction, greatly relaxing the technical requirements of the standard multicoil methods. Hardware and software implementations allow reduction of the reconstruction requirement to that of a single coil. The hardware implementation additionally allows a significant reduction in the net sampling bandwidth. The method is applicable to high speed MRI techniques, as demonstrated in phantoms and volunteers. PMID- 9402203 TI - MR microscopy of perfused brain slices. AB - To study the origins of signal changes in clinical MRI we have previously studied isolated single neuronal cells by MR microscopy. To account for the extracellular environment of the cells, we have developed a prototype perfusion chamber for MR microimaging of perfused rat hippocampal brain slices. To demonstrate the utility of this model, brain slices were initially perfused in isotonic solutions and then subjected to osmotic perturbations via perfusate exchange with 20% hypertonic and 20% hypotonic solutions. In diffusion weighted images, signal intensity changes of +16(sigma(n-1) = 11)% (hypotonic) and -26(sigma(n-1) = 10)% (hypertonic) were observed. No significant variation in response was observed across the slice when several subregions were examined. These observations are consistent with the view that contrast changes are driven primarily by changes in the intra- and extracellular compartmentation of water. This is the first report of MR microimaging of the isolated brain slice. The technique will enable the correlation of MR microimaging measurements with microscopic changes using other modalities and techniques to provide a better understanding of signals in clinical MRI. PMID- 9402204 TI - Elimination of eddy current artifacts in diffusion-weighted echo-planar images: the use of bipolar gradients. AB - Small gradient fields resulting from incompletely canceled eddy currents can cause geometric distortion in echo-planar images. Although this distortion is negligible in most echoplanar applications, the large gradient pulses used in diffusion-weighted echo-planar imaging can result in significant image distortion. In this report, it is shown that this distortion can be significantly reduced by the application of bipolar gradient waveforms. Both bipolar diffusion sensitizing gradients and an inverted gradient preparatory pulse were examined for minimizing the eddy currents responsible for these distortions. PMID- 9402205 TI - Spectrally weighted twisted projection imaging: reducing T2 signal attenuation effects in fast three-dimensional sodium imaging. AB - A scheme for the reduction of T2 signal attenuation effects in three-dimensional twisted projection imaging is presented. By purposely reducing the sample density at the high spatial frequencies, a considerable reduction in readout time is achieved. The reduction in readout time leads to decreased T2 signal attenuation which translates into improved signal-to-noise ratio (SNR). The SNR improvement is achieved without decreasing the image's resolution since the point spread function depends on the sample weighting as well as the T2 attenuation. The results indicate that SNR improvements of up to 40% can be achieved using the proposed scheme. PMID- 9402206 TI - k-Space detection and correction of physiological artifacts in fMRI. AB - Signal phase variations caused by physiology are a major source of instability in fMRI images produced by multiple RF pulses. k-Space phase variation maps show cyclic phase variations at the frequency of respiration combined with a cardiac variation of lower amplitude. The amplitude of the variation increases with gradient echo time and proximity to the chest, suggesting that the dominant cause of the phase variation is a B0 shift (approximately 0.01 ppm) produced by movement of organs in the chest. A simple k-space phase correction method is proposed and demonstrated for FLASH fMRI. The retrospective method requires no pulse sequence modification, and is more effective than navigator echo correction. Physiological noise is dramatically reduced, especially at inferior slice locations. PMID- 9402207 TI - Failure. PMID- 9402208 TI - The antimicrobial dance. PMID- 9402209 TI - The search for the Holy Grail: a century of anterior cruciate ligament reconstruction. AB - A perspective on the history of the development of anterior cruciate ligament reconstruction is presented. The lack of critical analyses establishing the relative effectiveness of many previously described procedures is documented. PMID- 9402210 TI - Bioresorbable fracture fixation in orthopedics: a comprehensive review. Part II. Clinical studies. AB - Bioresorbable materials overcome two major disadvantages of the metal alloys most commonly used in fracture-fixation devices: their extreme stiffness, which causes stress shielding of the underlying bone, and the necessity, in a significant number of cases, of removing metallic implants after fracture healing is complete. The orthopedic surgeon now has the use of polylactic acid, polyglycolic acid, and polydioxanone implants for the fixation of small cancellous bone fractures. The currently available bioresorbable materials lack strength and stiffness and are associated with inflammatory reactions and osteolysis in a significant number of cases. Surgeons should use the available pins and screws with extreme care and attention to the characteristics of each individual injury, particularly its healing characteristics, as well as to the material's initial mechanical properties, degradation rates, and associated complications. PMID- 9402211 TI - Treatment of grade III acromioclavicular separations in professional throwing athletes: results of a survey. AB - Forty-two team orthopedists representing all 28 major league baseball teams were surveyed to ascertain their definitive treatment for a hypothetical starting rotation pitcher who had sustained a grade III acromioclavicular (AC) separation to his throwing arm 1 week before the season. Twenty-nine (69%) of the physicians would treat the injury nonoperatively, while 13 (31%) would operate immediately. Twenty-five (60%) of the orthopedists had actually treated a pitcher or position baseball player with a grade III AC separation in the throwing arm, the 25 treating a total of 32 patients. Twenty (63%) of these injuries were treated nonoperatively, and 12 (37%) were treated operatively. The physicians reported that 16 (80%) of the patients treated nonoperatively regained normal function and achieved complete relief of pain, while 18 (90%) had normal range of motion after treatment; of those treated operatively, 11 (92%) regained normal function, achieved complete relief of pain, and had normal range of motion after surgery. PMID- 9402212 TI - Anatomic variations of the musculocutaneous nerve in the arm. AB - To determine the course and anatomic relationships of the musculocutaneous nerve in the arm, we dissected 54 cadaver arms and measured the length of any interconnection between the musculocutaneous nerve and the median nerve (36% of dissections; mean, 1.77 cm) and the distance from the coracoid process to (1) the musculocutaneous nerve (mean, 0.46 cm distal); (2) the median nerve (mean, 1.91 cm distal); (3) the musculocutaneous nerve's entrance to and exit from the coracobrachialis muscle (mean, 4.99 cm and 7.55 cm, respectively); and (4) the musculocutaneous nerve's entrance into the biceps muscle (mean, 11.66 cm). The high percentage of anomalies found emphasizes the complexities and irregularities of this anatomic region with regard to surgical approaches. PMID- 9402213 TI - Propagation of a patellar stress fracture in a basketball player. AB - Cyclic loading of cortical bone with repetitive, low-intensity loading can result in damage at its microstructural level. If this damage accumulates over time without appropriate repair, the strength of bone is reduced. Accumulation of fatigue damage may be manifest as stress fractures. As a link in the extensor mechanism of the leg, the patella is subject to complex loading, which varies with knee flexion and extension. Stress fractures of the patella originate on its anterior cortex and rarely propagate to completion. PMID- 9402214 TI - Paraspinal pyomyositis, a rare cause of severe back pain: case report and review of the literature. AB - Pyomyositis, a pyogenic infection of skeletal muscle, is rarely reported in temperate climates. A case of pyomyositis within the paraspinal muscles of a 63 year-old man is reported, with details of diagnostic evaluation and medical and surgical treatment of the condition. Failure to recognize this clinical entity can lead to diagnostic delay and inappropriate management. PMID- 9402215 TI - Bilateral femoral neck stress fractures in an amenorrheic athlete. AB - Stress fractures in athletes rarely involve the femoral neck. This report described the diagnosis and treatment of bilateral femoral neck stress fractures in a 30-year-old amenorrheic triathlete who is lactose intolerant and has a low caloric intake. The possibility of fatigue fracture should be considered in patients who have pain in the lower extremities that is exacerbated by activity, especially if they have hormonal or nutritional disorders. PMID- 9402216 TI - Syndesmotic ankle injuries in rodeo bull riders. AB - This report describes five male rodeo bull riders, aged 18 to 32 years, who sustained syndesmotic ankle injuries during competition. The mechanism of injury was identical in each case. As the rider attempted to escape after being thrown, the bull stepped on his lateral ankle, resulting in forced external rotation. All patients delayed seeking medical care, the interval between the injury and presentation ranging from 2 days to 4 weeks. All five patients sustained significant syndesmotic tears, two with associated fibular fractures. Three patients underwent surgical stabilization, and all riders returned to competitive rodeo events, often prior to medical clearance. To the authors' knowledge, this is the first reported association between injuries caused by livestock stepping and syndesmotic disruptions. PMID- 9402217 TI - Hyphenated history: the Hueter-Volkmann law. AB - The language that orthopedic surgeons speak is filled with eponyms. Perhaps more than any other medical specialty, we speak cryptically to one another using code words and secret language. Certain hyphenated eponyms are particularly interesting, because they pique one's curiosity as to how these persons came to be partners in orthopedic history. We will offer some bits of orthopedic hyphenated history, outlining the pertinent work of the respective authors as well as associated background information. This paper focuses on the Hueter Volkmann law, an important orthopedic concept concerning bone growth. This law bears the names of the two men credited with enunciating and popularizing it, Carl Hueter and Richard von Volkmann. PMID- 9402218 TI - Radius osteotomy assisted by temporary fixation with the Agee-Wristjack. AB - A modified technique for performing a distal radius osteotomy, using the Agee WristJack for temporary fixation, is described. The preoperative and postoperative conditions of a patient, a 26-year-old woman, are illustrated in radiographs. PMID- 9402219 TI - AMPA/kainate receptors modulate the survival in vitro of embryonic brainstem cells. AB - This study aimed at analyzing the involvement of (RS)-alpha-amino-3-hydroxy-5 methyl-4-isoxazolepropionic acid/kainate (AMPA/kainate) receptors in the survival of cultured rat embryonic brainstem cells, dissociated on embryonic day 14. The cell number was estimated after pharmacological manipulation of the receptors by exposure to agonists or antagonists. The developmental stage at the moment of drug application was critical for cell survival. We observed after 8 days in vitro a much stronger decrease in the number of gamma-enolase-positive cells when the cultures were treated for 3 days with the antagonist 6,7-dinitroquinoxaline 2,3-dione (DNQX) starting on the day of plating than when DNQX was added after 5 days in vitro. Conversely, exposure to the agonists (RS)-2-amino-3-(3-hydroxy-5 tri-fluoromethyl-4-isoxazolyl)-propion ic acid (T-AMPA) or kainate for 3 days significantly reduced cell survival only when the treatment was initiated after 5 days in vitro. Survival of S-100-positive cells was not affected after exposure to either agonists or antagonists. Neither agonist nor antagonist treatment modified cell proliferation, as assessed by 5-bromo-2'-deoxyuridine (BrdU) staining, suggesting that the decrease in the number of gamma-enolase-positive cells is essentially due to cell death. If some of the processes we observed in vitro correspond to analogous events in vivo, then exposure to excitatory amino acid receptor agonists or antagonists at critical stages of embryogenesis may alter the development of the central nervous system. PMID- 9402220 TI - Effect of a long-term nerve growth factor treatment on body weight, blood pressure, and serum corticosterone in rats. AB - Nerve growth factor is a well-characterized neurotrophin essential for the development and maintenance of certain central and peripheral neurons. As many neurons affected by aging depend for their survival on a constant supply of neurotrophins, nerve growth factor has been proposed as a possible treatment to prevent aging-associated neurodegeneration. There is evidence that nerve growth factor also plays a role in the immune system and modulates certain aspects of endocrine function. Here we have determined the effects of prolonged peripheral (intraperitoneal) treatment with nerve growth factor on body weight, blood pressure, and serum corticosterone levels in the rat. Our data indicate that intraperitoneally-injected nerve growth factor can affect body weight gain in rats. This effect may not be mediated by nerve growth factor-induced increases in serum corticosterone levels, as exogenous administration of corticosterone did not result in a similar body weight loss. These results show that, as previously reported for intracerebroventricular treatment with nerve growth factor, intraperitoneally-injected nerve growth factor also reduces body weight gain in rats. The data also suggest that exogenous delivery of nerve growth factor as part of therapeutic regimens is likely to have several effects. PMID- 9402221 TI - Cytokine regulation of embryonic rat dopamine and serotonin neuronal survival in vitro. AB - Interleukin-1beta (IL-1beta), interleukin-6 (IL-6), and tumor necrosis factor alpha (TNF-alpha) are cytokines with pleiotropic effects in the central nervous system (CNS), including an emerging role in neurodevelopment. This study measured the effects of cytokines on the survival of tyrosine hydroxylase (TH) immunoreactive dopamine neurons from the substantia nigra (SN), and 5 hydroxytryptamine (5-HT) immunoreactive serotonin neurons from the rostral raphe (RR), using cultures from embryonic day 14 (E14) rat brain. IL-1beta, IL-6, and TNF-alpha were added to cell cultures at 1, 10 and 100 U/ml. After 3 days in vitro, TH and 5-HT neurons were counted. The survival of 5-HT neurons was significantly reduced by 20-30% at 10 U/ml of IL-6. IL-1beta and TNF-alpha at doses of 1 and 10 U/ml appeared to have a similar effect on the survival of these neurons, but this effect was not statistically significant. Comparable non significant reductions of survival also occurred for TH neurons at the lower doses of IL-6 and TNF-alpha. In separate experiments, SN and RR cultures were exposed to the cytokines at a higher dose (1000 U/ml), causing a significant 30 40% decrease in the survival of TH neurons, but little or no change in 5-HT neuronal survival. Taken together, these results show that IL-1beta, IL-6, and TNF-alpha can affect developing monoamine neurons at physiologically relevant concentrations, and that high doses differentially inhibit the survival of TH and 5-HT neurons after short exposures. PMID- 9402222 TI - Fetuin expression in the dorsal root ganglia and trigeminal ganglia of perinatal rats. AB - Fetuin, a fetal plasma glycoprotein, has been shown previously to be present in sub-populations of neurons in the developing central and peripheral nervous system. To gain a more complete description of the time course of the appearance of fetuin during neurogenesis we have examined fetuin immunoreactivity, and the presence of fetuin mRNA, in the developing rat trigeminal and dorsal root ganglia. Fetuin immunoreactivity and its mRNA were first seen at embryonic day 15 in the trigeminal ganglia, and at embryonic day 16 in dorsal root ganglia. In both trigeminal and dorsal root ganglion, fetuin appeared to be present up until around the time of birth, and then again between postnatal days 3 and 16. The results suggest that fetuin first appears at around the time that ganglion cell axons reach their central targets, which is also approximately when the cell death period begins. The proportion of ganglion neurons that were fetuin immunoreactive at different ages was inversely related to the amount of cell death that is known to occur in these populations, thus it seems that fetuin is more likely to be associated not with dying cells, but with those that survive the cell-death period. PMID- 9402223 TI - Development of serotoninergic chick retinal neurons. AB - Numerous neurotransmitters have been studied in detail in the developing retina. Almost all known neurotransmitters and neuromodulators were demonstrated in vertebrate retinas using formaldehyde-induced fluorescence, uptake autoradiography or immunohistochemistry procedures. Serotoninergic (5HT) amacrine neurons were described in the inner nuclear layer (INL) of the retina with their dendrites spreading within the inner plexiform layer (IPL). The present work describes the morphological pattern of development of serotoninergic amacrine neurons with a stratified dendritic branching pattern in the chick retina from embryonic day 12 to postnatal day 7. Serotoninergic-bipolar neurons are also described. SHT-amacrine neurons have round or pear-shaped somata and primary dendritic trees oriented toward the IPL that runs through the INL, showing several varicosities. Secondary dendrites then go through the INL, without any collateral branch. At the outer and inner margin of the IPL the primary and secondary dendrites originate an outer and an inner serotoninergic network, respectively. When the primary dendritic tree reaches the IPL it deflects laterally in sublayer 1-the outer serotoninergic network. Tertiary branches then arise from the secondary dendrite and deflect in the innermost sublayer of the IPL-the inner serotoninergic network. The final pattern of branching of 5HT amacrine cells was present at embryonic day 14 and was completely developed at hatching. Serotoninergic (5HT) bipolar neurons were also present in the INL at hatching. They are weakly immunoreactive and are probably a subset of bipolar cells that accumulate serotonin from the intersynaptic cleft and are not "true" 5HT neurons. PMID- 9402224 TI - Age-dependent changes of presynaptic neuromodulation via A1-adenosine receptors in rat hippocampal slices. AB - The presynaptic neuromodulation of stimulation-evoked release of [3H] acetylcholine by endogenous adenosine, via A1-adenosine receptors, was studied in superfused hippocampal slices taken from 4-, 12- and 24-month-old rats. 8 Cyclopentyl-1,3-dimethylxanthine (0.25 microM), a selective A1-receptor antagonist, increased significantly the electrical field stimulation-induced release of [3H]-acetylcholine in slices prepared from 4- and 12-month-old rats, showing a tonic inhibitory action of endogenous adenosine via stimulation of presynaptic A1-adenosine receptors. In contrast, 8-cyclopentyl-1,3 dimethylxanthine had no effect in 24-month-old rats. 2-Chloroadenosine (10 microM), an adenosine receptor agonist decreased the release of [3H] acetylcholine in slices taken from 4- and 12-month-old rats, and no significant change was observed in slices taken from 24-month-old rats. In order to show whether the number/or affinity of the A1-receptors was affected in aged rats, [3H]-8-cyclopentyl-1,3-dimethylxanthine binding was studied in hippocampal membranes prepared from rats of different ages. Whereas the Bmax value was significantly lower in 2-year-old rats than in younger counterparts, the dissociation constant (Kd) was not affected by aging, indicating that the density rather than the affinity of adenosine receptors was altered. Endogenous adenosine levels present in the extracellular space were also measured in the superfusate by high performance liquid chromatography (HPLC) coupled with ultraviolet detection, and an age-related increase in the adenosine level was found. In summary, our results indicate that during aging the level of adenosine in the extracellular fluid is increased in the hippocampus. There is a downregulation and reduced responsiveness of presynaptic adenosine A1-receptors, and it seems likely that these changes are due to the enhanced adenosine level in the extracellular space. PMID- 9402225 TI - Transynaptic modulation by insulin-like growth factor I of dendritic spines in Purkinje cells. AB - Purkinje cells synthesize insulin-like growth factor I and express insulin-like growth factor I receptors during their entire life. An additional source of insulin-like growth factor I for these cells is provided by climbing fiber afferents originating in the inferior olive nucleus. Recently we found that insulin-like growth factor I from the inferior olive is necessary for motor learning processes probably involving Purkinje cell synaptic plasticity. We now studied whether inferior olive insulin-like growth factor I influences the synaptic structure of Purkinje cells, because changes in synaptic morphology are related to neuronal plasticity events. We injected an insulin-like growth factor I antisense in the inferior olive of adult rats, a procedure which we previously found to elicit a significant and reversible decrease of insulin-like growth factor I levels in the contralateral cerebellum. Ultrastructural analysis of the cerebellar cortex of these animals showed a significant reduction in the size of dendritic spines on Purkinje cells of antisense-treated rats compared to controls. The decrease in spine size was linked to a diminished numerical density of dendritic spines on Purkinje cells, without affecting the numerical density of synapses in the molecular layer of the cerebellum. This reduction was not due to a change in the thickness of the molecular layer. Climbing or parallel fiber terminals were also unaffected. Taken together with previous findings, these results support a role for insulin-like growth factor I produced in the inferior olive in the maintenance of Purkinje cell synaptic plasticity. PMID- 9402226 TI - Neuronal changes in the basolateral complex during development of the amygdala of the rat. AB - Neuronal changes in the amygdala basolateral complex were studied during development and maturation in fetal and postnatal rat brains using morphometrical methods. Forty brains of animals of various ages were fixed in formalin, frozen and cut into 25 microm thick sections and stained with cresyl violet or haematoxylin and eosin (H&E). In cresyl violet preparations, the complex appeared for the first time on embryonic day (E)17 and was composed of two homogeneous nuclei lateral and basolateral. On about the seventh postnatal day, each of these nuclei was divided into two parts the first one into the dorsolateral and ventromedial and the second one into the anterior and posterior. Morphometric investigations showed a different increase of the neuronal and nuclear size in various parts of the basolateral complex up to postnatal day (P)14; after that time these parameters did not change significantly. The neuronal density and the total number of neurons stabilized at P7 in all parts of this complex, except for the dorsolateral part of the lateral nucleus in which a 30% decrease of the total number of cells was observed. From P14, in all nuclei under study, the total number of neurons did not change significantly. PMID- 9402227 TI - Cloning, characterization and developmental expression of alpha2,8 sialyltransferase (GD3 synthase, ST8Sia I) gene in chick brain and retina. AB - GD3 and GM2 synthases act on ganglioside GM3 at the branching point of the pathway of synthesis of gangliosides in which the "a", "b" and "c" families are produced. The relative activities of these enzymes are important for regulating the ganglioside composition of a given tissue. In the present work, we report the cloning and characterization of a chick GD3 synthase cDNA. The cloned cDNA directed the synthesis of a functionally active enzyme in transiently transfected CHO-K1 cells and was highly homologous to mammalian GD3 synthases. In Northern blot experiments the cDNA detected a single specific GD3 synthase mRNA of about 9.0 kb both in the chicken brain and retina. The abundance of the specific mRNA transcript declined steadily from E7-E9 to very low values around PN2. The levels of enzyme activities measured at the same developmental stages roughly followed the changes of specific mRNA levels in both tissues. In situ hybridization of embryonic neural retina cells in culture showed that both glial- and neuron-like cells expressed the specific GD3 synthase mRNA, although with different intensities. Results indicate that transcription and/or stability of the specific GD3 synthase mRNA constitute a level of control of the expression of GD3 synthase and indirectly of the ganglioside composition in the developing chicken central nervous system (CNS). PMID- 9402228 TI - Valproic acid suppresses G1 phase-dependent sialylation of a 65kDa glycoprotein in the C6 glioma cell cycle. AB - The influence of valproate on in vitro glycosylation events in C6 glioma has been investigated, as this major human teratogen restricts proliferation in the mid-G1 phase of the cycle and alters the prevalence and/or glycosylation state of cell surface glycoproteins with the potential to mediate cell-cell and cell matrix interactions critical to development. C6 glioma cultured continuously in the presence of 1 mM valproate exhibited a significant depression of exponential growth but attained confluency one day later, when the majority of cells entered the G1 phase of the cycle. Glycoprotein sialyltransferase, which exhibited a four fold increase during exponential growth and a small decrease at confluency, was markedly attenuated in valproate-exposed cells in a manner which was indirect. This was associated with an inhibition of transient alpha2,3 sialylation of a 65 kDa glycoprotein expressed maximally at 4 hr into the G1 phase of the cell cycle. This effect was cell-cycle phase-specific, as exposure of synchronized cells to valproate inhibited transient sialylation at 4 and 5 hr into the G1 phase. Inhibition of the 65 kDa glycoprotein sialylation by valproate is suggested to arise from impaired signal transduction preceding the eventual arrest by the drug at a 5-6 hr G1 phase restriction point. PMID- 9402229 TI - Differential effect of taurine and serotonin on the outgrowth from explants or isolated cells of the retina. AB - The sulfur amino acid taurine and the indoleamine serotonin increases and decreases, respectively, the outgrowth from goldfish retinal explants. Taurine seems to be acting, at least partially, through an increase in calcium fluxes, and the serotonin-inhibiting effect appears to be mediated by serotonin1A receptors and cAMP. Isolated cells of postcrush goldfish retina and of retina from 5-day-old rats were cultured in the presence of taurine or serotonin. In the goldfish, the classical morphology of postcrush ganglion cells was observed. An antibody against the glycoprotein Thy-1 labelled three types of cells in the cultures of goldfish retina. The number of cells outgrowing and the length of the main neurite was measured at 5 days in culture in both species. The number of cells presenting neurites was increased in the goldfish retina by the addition of taurine, and decreased by serotonin. However, the length of the neurites was unaffected by the addition of the modulators. In the rat, only a slight decrease in the number of cells outgrowing was observed in the presence of serotonin. The incorporation of [3H]thymidine was not modified after 5 days in culture in the presence of taurine or serotonin, either in the goldfish or in the rat retina. The antibody Thy 1.1 can label retinal cells of the goldfish in vitro, one of them being ganglion cells. The trophic effect exerted by taurine in the postcrush goldfish retina needs the integrity of the tissue favoring the interaction of cells and factors, because outgrowth increases in retinal explants, but not in isolated cells. PMID- 9402230 TI - In vitro pattern formation during neurogenesis in neuroectodermal progenitor cells immortalized by p53-deficiency. AB - In vitro neural differentiation was induced in a p53-deficient immortalized neuroectodermal progenitor cell line, NE-4C, by treatment with retinoic acid [K. Schlett and E. Madarasz (1997) J. Neurosci. Res. 47, 405-416]. Rearrangement of nestin filaments was an early marker of neuron-formation. The increase in neurofilament protein content was accompanied by a decrease in the expression of nestin filaments in induced precursors. Cells with astroglial features appeared with a delay of 4-5 days compared to the appearence of neurons. Future neurons were sorted out from the substrate-attached population of apparently non-induced cells. The sorting out of future neurons resembled the separation of neural precursors in vivo. The continuous changes in the shape and also in the position of the cells resulted in the formation of characteristic morphological patterns. On the basis of morphological changes, five characteristic stages of in vitro neural differentiation were distinguished. The analysis of the morphological changes revealed that cell-to-cell interactions played an essential role in the cell fate decision made by induced precursors. Our observations indicate that the NE-4C cell line can serve as an in vitro model to investigate some early steps of neurogenesis. PMID- 9402231 TI - Developmental pattern of plasminogen activator activity in chick optic lobe. AB - Plasminogen activators are serine proteases which play a key role in morphogenesis and tissue remodelling. Two different molecular types, tissue-type and urokinase-type, were identified and they were postulated to play a role in neural development. The developing chick optic lobe plays a central role in processing visual information. In previous studies we demonstrated the occurrence of high levels of plasminogen activator activity in this model. The aim of the present paper is to study the temporal pattern of expression of this activity and characterize the type of plasminogen activator expressed in the developing optic lobe. Using soluble fractions derived by ultracentrifugation from Triton X-100 treated membrane fractions we measured the protease activity with a radial fibrinolytic assay. Employing different inhibitors of fibrinolytic activity and a zymographic assay, we showed that the developing optic lobe expresses only one type of plasminogen activator which corresponds to an urokinase-type of 70 kDa. Our results indicate that peaks of protease activity temporally correlate with massive neuronal migration, neurite outgrowth and synapse formation and maturation. This suggests that a plasminogen activator could play a role in these developmental events. This consistent pattern of variability strongly suggests that it is developmentally regulated and, if so, it could be a reliable parameter to study neural plastic changes induced by modifications in the environmental stimulation. PMID- 9402232 TI - Development of the tecto-thalamic projection neurons and the differential expressions of calcium-binding proteins in the rat. AB - We studied expression of calbindin-D 28 K and parvalbumin in tecto-thalamic projection neurons and during the formation of their tecto thalamic projections using a double-labeling with Fluoro-Gold. To discern the completion of these projections, Fluoro Gold, an opalescent fluorescent dye, was injected into the dorsal lateral geniculate and/or the lateral posterior nucleus in rats of various ages from neonates to adults. After one day's survival, the brains were removed and sections of the brain were immunohistochemically processed using Cy3, a red fluorescent dye, as a marker for calbindin-D 28 K or parvalbumin. The three types of tecto thalamic neurons, which have been described previously in the adult rats, were identified in the present study. The results revealed that in developing rats: 1) A population of medium-sized neurons (the presumed pyriform cells) express calbindin-D 28 K as early as the day of birth prior to the formation of their tecto thalamic projection that occured on postnatal day 4. Most (over 90%) of them project to the dorsal lateral geniculate nucleus; 2) A population of large neurons (the presumed wide-field vertical cells) express calbindin-D 28 K on postnatal day 7, and most of them (over 90%) project to the lateral posterior nucleus; 3) Another population of medium-sized neurons (the presumed narrow-field cells) express parvalbumin on post-natal day 17, but only a half (45%) of them project to the dorsal lateral geniculate nucleus. In the developing nervous system, calcium ions play important roles in the biological and molecular events underlying neural development. Changes in the free intracellular calcium ion level, indicating neuronal activity has been reported to be correlated with onset of calbindin-D 28 K or parvalbumin-immunoreactivity that participate in the regulation of intracellular calcium homeostasis in neurons. Therefore, the present findings may reflect distinct developmental events in the different classes of tectal relay neurons that form parallel visual pathways, but which have such different functions as the detection of luminance, discrimination of direction, and the detection of fast movements. PMID- 9402233 TI - An automated PACS image acquisition and recovery scheme for image integrity based on the DICOM standard. AB - The data quality and completeness of acquired images, which we refer to as integrity, is considered as the most important requirement in the image acquisition design of the Picture Archiving and Communication System (PACS). The Digital Imaging and Communications in Medicine (DICOM) standard significantly simplifies the task of acquiring radiological images from a DICOM compliant imaging system into the PACS. However, human interaction with the imaging system by changing the DICOM communication settings can result in missing images during the PACS image acquisition. A scheme based on the DICOM Query and Retrieve (Q/R) service class was developed to automatically identify and recover missing images. In addition, grouping sequential scanned images such as a CT and MR image series is another potential process that can miss images because of no indication of the end of series. Two methods are presented for determining the end of series and the pros and cons of each method are discussed in detail. Two experiments in a real clinical environment were conducted; one with and one without the Q/R implementation. The statistical results indicate two highlights from this work. First, the Q/R scheme faithfully recovered all missing images caused by human interaction with the DICOM compliant imaging system. Second, there was no single image slice missed when grouping slices into a series using the presented grouping algorithm in the two experimental periods. PMID- 9402234 TI - "Follow-up" method for processing of brain function MR images. AB - FMRI with standard 1.5 T scanners requires adapted algorithms because the time course of intensity signal showed a non-linearity of the baseline. The protocol contains sequential images covering periods of rest followed periods of stimulation. The images of each period of rest and stimulation were averaged, offering a series of averaged images. From this series, we conserved only the pixels which presented the alternated variations corresponding to the temporal pattern of the paradigm. A colour scale was used to present the average percentage of variations of each pixel selected. We have performed activation paradigms with a classical motor protocol. This simple "follow-up" method appears effective for the identification of activated areas. PMID- 9402235 TI - TAGASIST: a post-processing and analysis tools package for tagged Magnetic Resonance Imaging. AB - The Magnetic Resonance Imaging technique myocardial tagging allows the tracking and measurement of local parameters of heart wall motion. Tagging provides detailed information about regional cardiac function never before available with non-invasive techniques. The growth of this technique has been limited in part by the availability of post-processing tools. We introduce the TAGged cine AnalySIS Tools package (TAGASIST) for the processing of images from this technique. TAGASIST includes a graphical user interface for image segmentation, kinematic analysis, plotting of regional averages as well as multiple subjects (or patient populations) simultaneously, statistical analysis, and a database of all studies processed. PMID- 9402236 TI - Maxillary sinus inflammatory disease: ultrasound compared to computed tomography. AB - Fifty-six patients (age range, 15-79 yr, average, 37.0+/-18.5 yr), with a clinical and/or radiological diagnosis of acute maxillary sinusitis, were prospectively studied with ultrasound (US) and computed tomography (CT). The imaging finding which supported the diagnosis of acute sinusitis with US was the identification of the hyperechoic posterior antral wall through the hypoechoic inflammation. The findings were compared to CT (3 mm axial sections). The sensitivity of US for maxillary sinus disease was found to be 66.7% and the specificity was 94.9%, which were similar to the plain film ones (65.2 and 96.8%, respectively). The results of the present study suggest US as the method of first choice for acute sinusitis of the maxillary antra, particularly for children and pregnant women. PMID- 9402237 TI - Dynamic MR imaging of splenic tumor. AB - Inflammatory pseudotumor and hemangioma of the spleen are rare benign tumors, and MRI findings of splenic diseases have been reported only rarely. We recently observed three patients with inflammatory pseudotumor and hemangioma of the spleen. Abdominal ultrasonography, computed tomography (CT), magnetic resonance imaging (MRI) and angiography demonstrated within the enlarged spleen. MRI and dynamic MRI after administration of gadolinium DTPA provide the characterization of the splenic tumor. PMID- 9402238 TI - Invasive adenocarcinoma of the uterine cervix: MR imaging. AB - The detection of adenocarcinoma of the cervix is difficult in clinical examination, leading to poor prognoses. We evaluated MR appearances of 23 patients with adenocarcinoma of the cervix and appearances of 65 patients with cervical SCCs were also evaluated for comparison. Both T2-weighted spin echo, dynamic and postcontrast Gd-DTPA enhanced techniques were obtained at a 1.5 Tesla MR unit. The results of our study showed that adenocarcinoma of the cervix have MR imaging features which may permit distinction from SCCs which include: large tumors with preservation of endocervical epithelium, lesser contrast enhancement, and greater enhancement of the tumor periphery. Understaging may be more problematic with adenocarcinoma than SCC. PMID- 9402239 TI - Multiple, small, intracranial arachnoid cysts. AB - Arachnoid cysts are CSF-filled intraarachnoidal collections. They are most commonly located in the middle cranial fossa followed by suprasellar and quadrigeminal cisterns, posterior fossa, cerebral convexities, and interhemispheric fissure. We report an incidental, exceptional case with the coexistence of multiple arachnoid cysts in five different locations including two in the middle cranial fossae, and the other three in the retroclival region, quadrigeminal cistern, and cerebral convexity. PMID- 9402240 TI - Definitive diagnosis of enzymatic deficiencies of steroidogenesis in at-risk newborns and infants by urinary marker analysis using GC/MS-SIM. AB - A simplified urinary marker analysis for diagnosis of congenital adrenal hyperplasia (CAH) and 5alpha-reductase deficiency in infancy by GC/MS-SIM is introduced. The analysis was performed in 161 patients aged 3-90 days, 99 females and 62 males. CAH due to 21-hydroxylase deficiency was diagnosed in 61 patients (42 females and 19 males; in 10 cases simple virilizing form and in 51 patients salt-wasting form) and CAH induced by 3beta-hydroxysteroid dehydrogenase deficiency without salt loss in 1 female patient. In 2 full-term newborns and 6 preterm infants, a false-positive diagnosis of CAH, which had been based on serum steroid evaluation, was made. In these cases, increased excretion of fetal adrenal zone steroids was confirmed as a possible source of false-positive serum 11-deoxycortisol and 17alpha-hydroxyprogesterone values. Lack of fetal adrenal zone steroid metabolites in 2 male newborns with salt loss symptoms led to the diagnosis of adrenal insufficiency due to X-linked adrenal hypoplasia and adrenal hemorrhage. A single analysis of urinary CAH markers by the very sensitive and selective GC/MS-SIM method can replace numerous assays of various steroids that must be carried out for positive diagnosis of abnormal steroidogenesis in infancy. PMID- 9402241 TI - Measurement of serum exon 3-retaining growth hormone-binding protein in children and adolescents by radioimmunoassay. AB - Recently described assays for the determination of growth hormone-binding protein (GHBP) show a wide variety of normal ranges. Their results depend on the assay design and in the case of ligand-immunofunctional assay (LIFA), probably also on the binding characteristics, i.e. epitope specificity and affinity of the employed antibody. These facts underline the necessity to look for more accurate and specific assays. In this report we describe an accurate and simple radioimmunoassay (RIA) which allows the specific quantitation of the exon 3 retaining GHBP isoform (E3-GHBP). Data of the E3-GHBP RIA were compared to those of a LIFA measuring undifferentiated functional forms of GHBP. Our results demonstrate significant relationships between GHBP and age, BMI and IGF-I as determined by RIA and by LIFA in normal children and adolescents (n = 115, p < 0.001). Moreover, BMI is the only regulating factor of GHBP for both methods as shown by multiple regression analysis (p < 0.001). All our data suggest a qualitatively paralleled regulation of E3-GHBP and undifferentiated functional GHBP forms. This finding was confirmed by a good correlation between RIA and LIFA data (r = 0.74, p < 0.001). Children with idiopathic short stature (ISS, n = 47) had significantly lower GHBP levels than normal controls (n = 58) measured by the E3-GHBP RIA (p < 0.0001) and by LIFA (p < 0.01). We conclude that (1) ISS children may have a structural or quantitative defect at the level of the GHR, and (2) the highly specific assay for E3-GHBP immunoreactivity provides a sensitive diagnostic tool in conditions with partial GH insensitivity. PMID- 9402242 TI - Growth before and during growth hormone treatment in children operated for craniopharyngioma. AB - Height and weight growth before and during treatment with human growth hormone (hGH) was studied in 46 Dutch patients treated for craniopharyngioma. Weight was expressed as body mass index (BMI, weight/height). At the time of tumor treatment mean +/- SD height standard deviation score (SDS) was -1.22 +/- 1.38 and BMI SDS was 0.56 +/- 1.32. The initial height SDS was inversely related to age (r = 0.38; p < 0.02). Before hGH treatment height SDS decreased to - 1.57 +/- 1.08 (p < 0.05) and BMI SDS increased to 1.54 +/- 1.58 (p < 0.005) during the first year after tumor treatment. Changes in height SDS correlated positively with basal prolactin (PRL) levels (r = 0.46; p < 0.05). Neither tumor localization nor treatment mode was related to changes in height SDS and BMI SDS. Forty patients were treated with hGH, started a median interval of 2.0 years after tumor treatment. At the time of the start of hGH treatment height SDS in these patients was -1.70 +/- 1.13, and BMI SDS was 1.44 +/- 1.79. During treatment with hGH, height SDS increased to -1.05 +/- 1.10 (p < 0.001) in the first, and to -0.80 +/- 1.04 (p < 0.001) in the second year. BMI SDS did not change during hGH therapy. In conclusion, there is a large variation in height SDS and BMI SDS at the time of initial presentation as well as during spontaneous growth after tumor treatment. Spontaneous growth is related to serum PRL concentrations. Treatment with hGH significantly increased height SDS during the first 2 years, whereas BMI SDS did not change. PMID- 9402243 TI - Androstenedione, dehydroepiandrosterone sulfate, and estradiol levels throughout female puberty: relation to height velocity. AB - Sex steroids are important contributors to the pubertal growth spurt. Both androgens and estrogens have been related to this moment of rapid growth, but the role of estrogens is thought to be the most important one. Since exogenous estrogens are not capable to induce an appropriate growth spurt in girls, there might be an additional contributing factor involved. In a recent pilot study of 32 healthy pubertal girls, we found that the peak height velocity (HV) is preceded by relatively high levels of dehydroepiandrosterone sulfate and androstenedione (delta4A) as compared with the end-pubertal level. In the present study we evaluated HV in relation to dehydroepiandrosterone sulfate and delta4A levels in 149 healthy girls of various Tanner stages. HV was correlated with delta4A and estradiol levels in Tanner stages I-III. These results suggest that, like estrogens, delta4A might be an important stimulator of the female growth spurt. PMID- 9402244 TI - Concomitant evaluation of plasma galanin and catecholamine levels during a cold pressor test in healthy human male and female subjects. AB - The neuropeptide galanin (GAL) is localized in the peripheral and central nervous systems as well as in the adrenal medulla where it coexists with catecholamines. We evaluated the changes in GAL plasma levels as well as systolic and diastolic blood pressures and in the plasma levels of epinephrine and norepinephrine (NE) in normal human male and regularly menstruating female subjects during the activation of the sympathoadrenal system by a cold pressor test. The test was performed by immersing the hand of the subject in 1 degree C cold water for 4 min. Blood samples were collected both under basal conditions and at subsequent intervals during the cold stimulus as well as at the end of the recovery phase. The values were compared with those obtained when the same subjects were sham tested. As expected, systolic and diastolic blood pressures increased in both sexes during the cold test; the systolic blood pressure values were significantly (p < 0.05) higher in males. Epinephrine and NE levels rose significantly above baseline in both male and female subjects after the cold stimulus; the NE increments were significantly (p < 0.05) higher in males. The basal GAL levels were found to be variable but not sexually dimorphic. In both sexes, during cold stimulus and recovery phase, GAL values were found to be not significantly different from those detected during the sham test. These results demonstrate that the release of GAL in peripheral blood is not associated with that of catecholamines in response to the cold pressor test. PMID- 9402245 TI - Final height after growth hormone therapy in short children: correlation with siblings' height. AB - Sixteen prepubertal children (10 males, 6 females) were treated with growth hormone (GH) 0.75 U/kg/week for 4-5 years until final height was attained. Before initiation of GH therapy height was below 2 SDS for age and gender, growth velocity was <4.5 cm/year, bone age was more than 2 SD below the mean for age, and the GH response to provocative tests was more than 10 microg/l. The final height of 9 patients exceeded their predicted and target heights. The final height of the treated patients was highly correlated with their siblings' heights (r = 0.806, p < 0.001) and to a lesser degree with the target or the predicted height (r = 0.401, p = 0.124 and r = 0.465, p = 0.06, respectively). It is concluded that the secular trend should be taken into consideration when evaluating the success of GH therapy. PMID- 9402246 TI - Surface changes of ram spermatozoa by adsorption of homologous and heterologous seminal plasma proteins revealed by partition in an aqueous two-phase system. AB - Ram spermatozoa freed from seminal plasma by a dextran 'swim-up' procedure were incubated with Tween 20 and fractionated into motile (PEG-rich) and stationary (dextran-rich) fractions by centrifugal countercurrent distribution (CCCD) in an aqueous dextran-Ficoll-polyethylene glycol (PEG) two-phase system. Increasing concentrations of Tween 20 resulted in greater amounts of extracted protein and lower cell viability. Addition of bull seminal plasma increased the proportion of live cells, whereas ram seminal plasma increased the proportion of stationary cells. Proteins isolated from each type of seminal plasma restored the CCCD profile of treated spermatozoa to the right, this effect being reduced when proteins were thermally denatured. Bovine serum albumin induced a slight displacement to the left. No restoration of profile was achieved when ram spermatozoa were exposed to proteins from bull seminal plasma in the presence of protein-free ram seminal plasma. Adsorption of seminal plasma proteins by spermatozoa previously stripped of surface proteins by exposure to detergent reversed the detergent effect on motility. The findings are consistent with the concept that ram seminal plasma contains a factor that interferes with protein adsorption on the cell surface and prevents the protective effect of seminal plasma proteins on maintenance of cell viability. PMID- 9402247 TI - Nitric oxide mediates human chorionic gonadotrophin-induced prostaglandin E generation in rat oocyte-cumulus complexes. AB - To determine whether nitric oxide (NO) generation mediates human chorionic gonadotrophin (hCG)-induced prostaglandin E (PGE) secretion by oocyte-cumulus complexes (OCC), the secretion of PGE by cultured rat OCC in the presence of NO donors and NO synthase (NOS) inhibitors was characterized. NO donors (sodium nitroprusside and 3-morpholino-sydnonimine-hydrochloride) increased PGE accumulation in OCC to values similar to those obtained in the presence of hCG. The three NOS inhibitors tested (NG-nitro-L-arginine methyl ester, NG-monomethyl L-arginine and aminoguanidine) prevented the hCG-induced PGE accumulation in cultured OCC. This effect appears to be specific since D-enantiomers NG-nitro-D arginine methyl ester and NG-monomethyl-D-arginine had no effect. The present results suggest that NO mediates the hCG-induced accumulation of PGE in rat OCC, a process which may occur in vivo in preovulatory follicles prior to ovulation. PMID- 9402248 TI - Characterization and proteolytic activity of a cathepsin L-like polypeptide in endometrium and uterine flushings of cycling, pregnant and steroid-treated ovariectomized gilts. AB - Cathepsin L has been proposed to be involved with the endothelial-chorial type of placentation in the cat. Little information concerning the presence and secretion of cathepsin L is available for a species with noninvasive epitheliochorial placentation such as the pig. Cathepsin L activity in uterine flushings and endometrium from gilts during different days of the oestrous cycle and early pregnancy was analysed through specific substrate metabolism and Western blot analyses with antiserum against cat endometrial cathepsin L. This antiserum was utilized to determine the cellular localization of the enzyme within porcine endometrium. Cathepsin L activity within uterine flushings was elevated on Day 15 of the oestrous cycle and early pregnancy, with activity declining on Day 18. Cat cathepsin L antiserum cross-reacted with a group of 46, 40 and 38 kDa uterine proteins and detected a product within the surface and glandular epithelium of the endometrium. The appearance of the 40 kDa protein was first detected on Day 10 of the oestrous cycle with the 38 kDa proteins appearing on Day 15 and 18 of pregnancy. The 40 and 38 kDa uterine proteins appear to be steroid regulated as 12 days of progesterone administration is necessary to detect the proteins and cathepsin L activity. PMID- 9402249 TI - Localization and synthesis of collagen types III and V during remodelling and decidualization in rat uterus. AB - Uteri of pregnant rats on Days 6, 7 and 8 of pregnancy were studied to determine the histochemical distribution of collagen types III and V and the incorporation of [3H]glycine into fibrillar collagens during the period of embryonic implantation. Types III and V had a similar distribution in the non-decidual stromal region and muscle layers in implantation sites. They were found to have very low levels in the primary decidual tissue on Day 6 and were not detected in developing decidual tissues on Days 7 or 8. Following injection of labelled glycine, collagen was extracted and the specific activity of the collagens determined by fluorography and 3H incorporated into the collagen bands in the gels. It was found that incorporation of label into both types I and III was similar (33.4+/-12.0 and 31.8+/-18.1 cpm microg-1 collagen respectively) but 3.5 times that of type V (7.7+/-5.3 cpm microg-1). These studies suggest that although fibrillar collagens are metabolized or redistributed in the growing decidual tissue, they are incorporated rapidly into the extracellular matrix during remodelling of the outer stroma and muscle tissues. PMID- 9402250 TI - Perifusion culture system for bovine embryos: improvement of embryo development by use of bovine oviduct epithelial cells, an antioxidant and polyvinyl alcohol. AB - Three experiments were conducted in an attempt to improve a continuous flow perifusion system capable of maintaining embryo development for long periods of time. Bovine embryos (8-16 cells) obtained from static co-culture with cumulus cells in a serum-free medium were perifused in an ACUSYST-S cell culture incubator. Culture chambers of the incubator consisted of a 0.2-mL unit (Chamber 1) connected to a 1.5-mL unit (Chamber 2), with the outflow from Chamber 1 routed to the inlet to Chamber 2. A bovine embryo culture medium supplemented with 3 mg mL-1 bovine serum albumin (BSA) and 25 mM HEPES was used as a perifusion culture medium (PCM). Embryos were perifused in Chamber 2 for 24, 48 and 72 h and further co-cultured in a static system up to 216 h after insemination. In Experiment 1, conditioning PCM with frozen-thawed bovine oviduct epithelial cells (BOEC) placed in Chamber 1 enhanced (P < 0.05) blastocyst formation of embryos in Chamber 2, after 24, 48 and 72 h of perifusion culture. The proportion of blastocysts was not further increased by placing BOEC in Chamber 2 along with the embryos. In Experiment 2, embryos were perifused with PCM conditioned with BOEC in Chamber 1 for 48 h or 72 h. A higher proportion of perifused embryos developed to the blastocyst stage after addition of 25 U mL-1 or 50 U mL-1 of superoxide dismutase (SOD) to PCM than in its absence. However, blastocyst formation of embryos perifused for 72 h was not increased after addition of 50 U mL-1 SOD compared with its absence. In Experiment 3, the proportions of morulae and blastocysts were not decreased by replacement of 3 mg mL-1 BSA with 1 mg mL-1 polyvinyl alcohol (PVA) in a BOEC-conditioned medium containing 50 U mL-1 SOD after perifusion for 48 h. In conclusion, PCM conditioning with BOEC and addition of an antioxidant to the perifusion medium improved the developmental capacity of perifused embryos. PVA is an adequate replacement for BSA in the perifusion medium. PMID- 9402251 TI - Isolation, characterization and radioimmunoassay of luteinizing hormone in the brushtail possum. AB - Luteinizing hormone (LH) was purified from brushtail possum (Trichosurus vulpecula) pituitary glands. The purification procedure consisted of ammonium sulfate precipitation followed by triazinyl-dye chromatography, hydrophobic interaction chromatography and gel filtration. A yield of 10 microg LH g-1 pituitary with a recovery of 20% was obtained from 1400 pituitary glands (20.3 g). Contamination with possum follicle-stimulating hormone (FSH) was < or =0.05%. The amino acid analysis and the N-terminal sequencing for 10 cycles revealed close homology with LH from other mammals. Minor amounts of LH that had been truncated near the N-terminal were also detected. No contaminating proteins were found by amino acid sequencing. The potency of possum LH was 20% that of ovine LH in a receptor assay using possum testicular receptors and 4% that of ovine LH when bovine corpora lutea receptors were used. Possum LH was able to stimulate production of cyclic adenosine 3',5'-monophosphate by bovine granulosa cells. A radioimmunoassay (RIA) for possum LH using 125I-possum LH and an antiserum raised against ovine LH was developed. The RIA has a sensitivity of 0.15 ng mL-1, a 50% displacement of 1.9 ng mL-1 and a cross-reactivity of <0.02% against possum FSH. Plasma concentrations were 0.24+/-0.04 ng mL-1 (n = 8) and 0.39+/-0.12 ng mL-1 (n = 8) in female and male possums respectively. Administration of mammalian gonadotrophin-releasing hormone (GnRH) and chicken GnRH II stimulated increases in plasma LH concentrations in male and female possums. When comparing LH responses with administration of mammalian GnRH or chicken GnRH II, plasma LH concentrations remained elevated for a longer period of time in males than in females (P < 0.01); plasma LH concentrations also remained elevated for longer after mammalian GnRH than after chicken GnRH II (P < 0.01). Gonadectomy stimulated an increase in plasma concentrations of LH in both male (P < 0.01) and female (P < 0.05) possums. The rate of increase in plasma LH concentrations in males was faster than that in females. In summary, we have purified, partially characterized, and developed a RIA for possum LH. PMID- 9402252 TI - Effects of pivalic acid and sodium pivalate on L-carnitine concentrations in the cauda epididymidis and on male fertility in the hamster. AB - In this study, administration of pivalic acid or its sodium salt was found to decrease the L-carnitine concentration in the epididymal lumen of the hamster; it also tested whether this decrease affected sperm cell motility, chromatin structure, or fertilizing capacity. Provision of pivalic acid or its sodium salt (20 mM or 40 mM) in the drinking water of mature male golden hamsters for 30 days reduced (by 72%, 75%, and 83% in three experiments) the L-carnitine concentration of the cauda epididymidis but did not inhibit sperm chromatin condensation, as assessed by flow cytometry. The treatments did not alter the location of motile sperm in the epididymidis nor did they appreciably affect the motility of sperm obtained from the distal cauda epididymidis. The numbers and percentage of ova that reached the 2-cell stage 36-40 h after uterine insemination with spermatozoa from control and treated hamsters served as a measure of sperm fertility. Treatment with pivalic acid or sodium pivalate did not render male hamsters infertile although it appeared to reduce the fertilizing ability of their spermatozoa. These results suggest that the high concentration of L-carnitine present in the lumen of the cauda epididymidis is not required for maturation of sperm chromatin or development of sperm motility. PMID- 9402253 TI - Evidence that nitric oxide synthase is involved in progesterone-induced acrosomal exocytosis in mouse spermatozoa. AB - In a recent work, we detected nitric oxide synthase (NO synthase) in the acrosome and tail of mouse and human spermatozoa by an immunofluorescence technique. Also, NO-synthase inhibitors added during sperm capacitation in vitro reduced the percentage of oocytes fertilized in vitro, suggesting a role for NO synthase in sperm function. Therefore, in the present study the effect of three NO-synthase inhibitors, NG-nitro-L-arginine methyl ester (L-NAME), NG-nitro-D-arginine methyl ester (D-NAME) and L-NG-nitro-arginine (NO2-arg), and of a nitric oxide donor, spermine-NONOate, on the progesterone-induced acrosome reaction of mouse sperm was examined. NO-synthase inhibitors were added at 0, 60 or 90 min during capacitation; at 120 min, mouse epididymal spermatozoa were exposed to 15 microM progesterone for another 15 min. In another set of experiments, different concentrations of spermine-NONOate were added to capacitated spermatozoa for 15 min; in these experiments, progesterone was not included. NO2-arg and L-NAME blocked progesterone-induced exocytosis regardless of the time at which these inhibitors were added. Moreover, D-NAME did not inhibit exocytosis. In contrast, spermine-NONOate stimulated the acrosomal exocytosis in vitro directly. These results provide evidence that mouse sperm NO synthase participates in the progesterone-induced acrosome reaction in vitro and that nitric oxide induces this event. PMID- 9402254 TI - Effect of cytochalasin B and cycloheximide on the activation rate, chromosome constituent and in vitro development of porcine oocytes following parthenogenetic stimulation. AB - Activation rate, chromosome constituent and developmental pattern of porcine oocytes was examined in the presence and absence of cytochalasin B and cycloheximide following parthenogenetic stimulation. Treatment with cycloheximide after ethanol or Ca2+ ionophore treatment increased the incidence of activation. The percentage of oocytes with two or more female pronuclei was higher (P < 0.05) in oocytes treated with cytochalasin B than in control or cycloheximide-treated oocytes. Treatment with both electrical stimulation and cytochalasin B increased the incidence of diploid chromosome spreads, and accelerated development to the morula and blastocyst stage compared with the control and cycloheximide-treated groups, suggesting a role of ploidy in the development of parthenote. PMID- 9402255 TI - Effects of passive immunization against oestradiol-17beta and inhibin on the secretion of gonadotrophin in the cyclic golden hamster (Mesocricetus auratus). AB - To investigate the physiological importance of oestradiol-17beta and inhibin in the regulation of gonadotrophin secretion in the cyclic golden hamster, females were passively immunized against two hormones. When 200 microL antiserum against oestradiol-17beta (oestradiol-AS) was given on Day 3 (Day 1 = day of ovulation), the preovulatory gonadotrophin surge was completely blocked for 24 h and the length of the oestrous cycle was also prolonged for one day. In the group given 200 microL oestradiol-AS on Day 3, basal levels of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) increased slightly and superovulation (19.6+/ 0.8, mean+/-s.e.m.) occurred. When 200 microL antiserum against inhibin (inhibin AS) was given at 1100 hours on Day 3, a dramatic increase in plasma FSH and a slight increase in LH were noted, resulting in superovulation (38.2+/-2.6) on the expected Day 1. The present study indicates clearly that inhibin plays a major role in regulating the specific ovulation rate in the hamster through the control of FSH secretion. Present results also indicate that oestradiol-17beta suppresses basal LH secretion. Oestradiol-17beta may act as an indicator of the follicular maturation, and the high plasma concentration of oestradiol-17beta noted from Day 3 to Day 4 may play an important role in determining the timing of initiation of the preovulatory gonadotrophin surge. PMID- 9402256 TI - Collagen in the fetal membranes of sheep: changes throughout gestation and effects of dexamethasone at 60 days. AB - The tensile strength of fetal membranes is largely due to their collagen content. In this study we have examined the changes in collagen in the amniotic and allantoic membranes of the sheep over a wide gestational range (27-142 days of gestation; term, 145-150 days). The results have been correlated with volume changes in normal development, and in particular, the changes in allantois have been studied after a rapid and extensive increase in allantoic volume, as a result of maternal dexamethasone treatment (0.76 mg h-1 for 48 h) from Day 60 of gestation. Electron microscopy and immunohistochemistry were used to delineate collagen distribution, and gel electrophoresis was used to assess the relative proportions of each type. In the amnion, collagen content increased from 37 +/- 4% to 50 +/- 1% dry weight of the tissue from 41-102 days and declined slightly thereafter. In the allantois, collagen content increased from 20 +/- 1% at Day 27 to 50 +/- 6% at Day 142, significantly correlated with a volume increase from 25 +/- 3 mL to 813 +/- 274 mL. Collagen types I (>85%), III (10%) and small amounts of types IV and V (<5%) were identified in both membranes at all ages. When allantoic fluid volume was increased rapidly by maternal dexamethasone infusion, there was a significant decrease in collagen content from 38 +/- 6% to 25 +/- 2% (P < 0.05). By immunohistochemistry it was observed that both epithelial cells and fibroblasts were synthesizing collagen. PMID- 9402257 TI - Male-dependent variability of fertilization and embryo development in two bovine in vitro fertilization systems and the effects of casein phosphopeptides (CPPs). AB - This study investigated the effects of semen from five different bulls and two different ejaculates of the same bull on penetration, cleavage, blastocyst formation, and cell allocation in bovine blastocysts produced in vitro. Casein phosphopeptides (CPPs) were tested for their ability to enhance fertilization and minimize variability among bulls and ejaculates. In Experiment 1, the BO fertilization system was employed. Penetration and polyspermy both displayed great variation among bulls and between ejaculates, whereas no significant differences were observed in cleavage and blastocyst-formation rates. Similar variability was observed in penetration, polyspermy, cleavage, blastocyst formation rates and cell allocation and distribution when the two fertilization systems, TALP and BO, were compared in Experiment 2. The BO-system supported penetration and polyspermy better (P < 0.05) than the TALP-system, whereas the TALP-system was superior (P < 0.05) in supporting cleavage and blastocyst formation. Significant interactions existed between bulls and the fertilization system employed. It is concluded that the success of in vitro fertilization is markedly dependent on individual bulls as well as on ejaculates from the same bull. CPPs are able to enhance penetration and embryo development in certain bulls or ejaculates and thus contribute to reducing the degree of individual variability, but they do not generally improve the success of bovine embryo production in vitro. PMID- 9402258 TI - Isolation and partial characterization of tammar wallaby luteinizing hormone and development of a radioimmunoassay. AB - Tammar wallaby (Macropus eugenii) luteinizing hormone (LH) was purified from pituitaries collected from wild and captive populations by salt sequential precipitation, ion exchange chromatography and gel filtration. Pituitary tissue (5 g) yielded 1.8 mg of purified wallaby luteinizing hormone (ME-14B), as verified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). A heterologous radioimmunoassay has been developed for measurement of LH in plasma of marsupials using a monoclonal antibody raised against bovine LH (518B7). This assay system was able to measure basal LH concentrations in male and female tammars and detected a significant rise in plasma LH in response to oestradiol benzoate in female tammars and luteinizing hormone-releasing hormone (LHRH) in males. Parallel dose-response curves were also obtained from pituitary extracts from four other species of marsupial (brushtail possum, Trichosurus vulpecula; brown antechinus, Antechinus stuartii; kowari, Dasyuroides byrnei; and Eastern pygmy possum, Cercartetus nanus) in this assay, which suggests its usefulness in the measurement of LH in other marsupial species. PMID- 9402259 TI - Why pre-eclampisa? PMID- 9402260 TI - From birds and bees to babies? Can theories on genetic conflict aid the clinician? PMID- 9402261 TI - Cervical ripening in guinea-pigs after a local application of nitric oxide. AB - In humans cervical ripening is an inflammatory reaction accompanied by an infiltration of white blood cells and the remodelling of the extracellular matrix. Similar changes occur in guinea-pigs during cervical ripening. Nitric oxide (NO) is thought to be important in the maintenance of pregnancy because it is synthesized by the uterus and inhibits contractility. Previous studies in rats also demonstrated that an NO generating system is present in the cervix and is up regulated during labour. We studied the local effect of the NO donor sodium nitroprusside (SNP) on cervical ripening in guinea-pigs during advanced pregnancy. SNP (5 mg/injection) was administered into the cervical canal in 0.2 ml phosphate-buffered saline containing 3% hydroxycellulose twice a day either for 1 [on day 42 post coitum (p.c.)] or 2 consecutive days (days 42-43 p.c.; term day 67 + 3 p.c.). The effects were assessed 24 h after treatment by both extensibility measurements (force resistance to incremental stretch) and morphological evaluation (light and electron microscopy after in-situ fixation). The control animals were treated with the vehicle. In another experiment, the guinea-pigs were subcutaneously (s.c.) treated on day 43 p.c. with either the progesterone antagonist onapristone (3 and 10 mg/animal s.c.) or with prostaglandin E2 (PGE2) (1 and 3 mg/animal s.c.) and the PGE2 analogue sulprostone (0.03 and 0.1 mg/animal s.c.). The cervical extensibility was measured 24 h later. One-day SNP treatment tended to reduce cervical resistance, but not significantly, whereas 2 day treatment with SNP led to a significant increase in cervical extensiblity (P < 0.05) with little effect on cervical dilatation (indirect evidence of the absence of uterine contractions). The effects on cervical resistance were comparable to those achieved with 10 mg onapristone and high-dose prostaglandins (PG)s (3 mg PGE2 and 0.1 mg sulprostone) treatment. An electron microscope study of the SNP-treated animals revealed a dissolution of collagen fibres, stromal oedema, arterial dilatation, and the infiltration of macrophages, lymphocytes and granulocytes. Numerous mast cells were also present. The morphological effects of SNP were similar to those observed during normal cervical ripening at term. We conclude that the local application of a NO donor effectively induces cervical ripening without inducing labour in pregnant guinea-pigs. PMID- 9402262 TI - Splenocytes in early pregnancy promote embryo implantation by regulating endometrial differentiation in mice. AB - We have reported that i.v. administration of splenocytes prepared from early pregnant mice promoted embryo implantation in pseudopregnant CD-1 (ICR) (closed colony) mice. In this study, the similar effects of splenocytes were confirmed using an inbred strain, BALB/c mice, and the mechanism was further investigated. Splenocytes were prepared from pregnancy day 4 (P4-spl) and dioestrus mice (Di spl), and supernatant of P4-spl suspension (P4-sup) was used as controls. On pseudopregnancy day 2, splenocytes or supernatant were injected into caudal vein or endometrial stroma of the recipient mice, and blastocysts were transferred into the endometrial lumen. In both BALB/c and ICR strains, the implantation rates per recipient with i.v. and intraendometrial injection were significantly higher in P4-spl groups. Then, ICR mice were oophorectomized on pseudopregnancy day 3. After 3 day progesterone supplementation, blastocysts were transferred with i.v. injection of P4-spl and P4-sup, or s.c. oestradiol injection. Under progesterone supplementation, successful implantations were observed in the P4 spl- and oestradiol-treated groups, but not in P4-sup-treated group. Reverse transcriptase-polymerase chain reaction analysis revealed that messenger RNA expression of leukaemia inhibitory factor in the uterus was induced by P4-spl and oestradiol, but not by P4-sup. These findings showed that splenocytes of early pregnant mice promote embryo implantation by regulating endometrial differentiation. PMID- 9402263 TI - Ontogeny of gonadotrophin and inhibin secretion in normal girls through puberty based on overnight serial sampling and a comparison with normal boys. AB - We measured luteinizing hormone (LH) and follicle stimulating hormone (FSH) by immunofluorometric assays and alpha-inhibin by radioimmunoassay in serum sampled every 10 min throughout the night (2100-0500 h) from 44 normal girls. Mean overnight LH values rose log-linearly from a mean of 0.2 IU/l in prepubertal girls to 3.0 IU/l in late pubertal girls. Log2 mean overnight FSH rose rapidly through early puberty and then remained constant; mean FSH rose from 1.0 IU/l in prepubertal girls to approximately 2.8 IU/l in Tanner III-V girls. Mean overnight inhibin increased through puberty, rising from 151 ng/l in prepubertal girls to 432 ng/l in fully pubescent girls. Within each of the first three Tanner stages, LH differed approximately 100-fold between the smallest and largest mean concentrations but differed <10-fold within stages IV or V. Such within-pubertal stage variability was less pronounced for FSH, which differed approximately 16 fold among Tanner I subjects and 4-10-fold at later stages, and for inhibin, which varied approximately 4-fold within each Tanner stage. The frequency of LH pulses during overnight sampling increased significantly during puberty, but the frequency of FSH and inhibin pulses remained constant. We compared the results from girls to those from 50 normal boys [Manasco et al. (1995) J. Clin. Endocrinol. Metab., 80, 20462052]. At each pubertal stage, girls had approximately the same mean overnight LH values as boys; girls had higher mean overnight FSH, particularly during Tanner stages II-IV; and boys had mean overnight alpha-inhibin immunoreactivity approximately 1.5 times that of girls at each pubertal stage. Still, hormone concentrations for individuals of both sexes intergraded at each pubertal stage. PMID- 9402264 TI - Increased concentrations of renin, aldosterone and Ca125 in a case of spontaneous, recurrent, familial, severe ovarian hyperstimulation syndrome. AB - We report for the first time increased concentrations of aldosterone and renin in a case of spontaneous, recurrent, familial, severe ovarian hyperstimulation syndrome (OHSS). High concentrations of Ca125 were also found. Our patient was a 26 year old woman, gravida 2, para 1, affected by severe OHSS, who denied having ever consumed any ovulation drug. Both the patient and her only sister had suffered from a similar condition in their previous pregnancies. The patient was treated with i.v. fluid therapy. Paracentesis was performed on one occasion. The patient was dismissed after 25 days in good condition. Blood count and blood chemistry confirmed the severity of the clinical picture. We conclude that spontaneous OHSS, although very rare, may have been underestimated so far. It can be recurrent and may also be familial. The intra-ovarian prorenin-renin angiotensin system may play a role in its aetiopathogenesis. PMID- 9402265 TI - Effect of disruption versus continuation of gonadotrophin-releasing agonist after human chorionic gonadotrophin administration on corpus luteum function in patients undergoing ovulation induction for in-vitro fertilization. AB - Previous studies have described the luteolytic effect of gonadotrophin-releasing hormone agonist (GnRHa) administered in the early luteal phase. The present work was undertaken to compare in a prospective and randomized design the effect of disruption versus continuation of daily GnRHa after human chorionic gonadotrophin (HCG) administration on corpus luteum function in patients undergoing ovulation induction for in-vitro fertilization (IVF). Two different studies were designed and a total of 38 ovum donors, aged 23-30 years, were included. In the first study, the effect of GnRHa on the early luteal phase of IVF-stimulated cycles was investigated (n = 27); the patients were divided into two groups, according to whether they stopped (n = 13) or continued with daily GnRHa injections (n = 14) for an additional period of 15 days after HCG administration. Blood was drawn from luteal phase days 2 to 6 (day 0 = day of HCG administration) and oestradiol and progesterone concentrations were analysed. The second study focused on the effects of continuation versus disruption of GnRHa administration in the mid-late luteal phase. A similar design was employed including six patients who stopped GnRH on day 0 and five other women who continued GnRHa for 15 days after HCG administration. In this second study, blood was drawn from days 5 to 11 and oestradiol, progesterone and luteinizing hormone (LH) concentrations were analysed. IVF parameters were similar in both groups. The results indicate that continuous GnRHa administration, after HCG injection, does not produce changes in oestradiol, progesterone and LH concentrations in the early, mid- and late luteal phases compared to those patients in whom GnRHa is discontinued at the day of HCG administration. The present work demonstrates that, when ovulation induction is performed, the corpus luteum is driven primarily by the HCG, regardless of the administration or disruption of GnRHa in the luteal phase. This suggests that the lack of differences between continuation versus disruption of GnRHa may be due to the accumulation of the product over the previous 2-3 weeks of treatment. PMID- 9402266 TI - Changes in serum concentrations of growth hormone, insulin, insulin-like growth factor and insulin-like growth factor-binding proteins 1 and 3 and urinary growth hormone excretion during the menstrual cycle. AB - Few studies exist on the physiological changes in the concentrations of growth hormone (GH), insulin-like growth factors (IGF) and IGF-binding proteins (IGFBP) within the menstrual cycle, and some controversy remains. We therefore decided to study the impact of endogenous sex steroids on the GH-IGF-IGFBP axis during the ovulatory menstrual cycle in 10 healthy women (aged 18-40 years). Blood sampling and urinary collection was performed every morning at 0800 h for 32 consecutive days. Every second day the subjects were fasted overnight before blood sampling. Follicle stimulating hormone, luteinizing hormone (LH), oestradiol, progesterone, IGF-I, IGFBP-3, sex hormone-binding globulin, dihydroepiandrosterone sulphate and GH were determined in all samples, whereas insulin and IGFBP-1 were determined in fasted samples only. Serum IGF-I concentrations showed some fluctuation during the menstrual cycle, with significantly higher values in the luteal phase compared to the proliferative phase (P < 0.001). Mean individual variation in IGF I concentrations throughout the menstrual cycle was 13.2% (SD 4.3; range 0.1 18.3%). There were no cyclic changes in IGFBP-3 serum concentrations and no differences in IGFBP-3 concentrations between the luteal and the proliferative phases. Mean individual variation in IGFBP-3 concentrations throughout the menstrual cycle was 8.8% (SD 2.7; range 3.2-14.1). IGFBP-1 concentrations were inversely associated with insulin concentrations, and showed a significant pre ovulatory increase that returned to baseline at the day of the LH surge. Fasting insulin concentrations showed large fluctuations throughout the menstrual cycle without any distinct cyclic pattern. No cyclic changes in urinary GH excretion during menstrual cycle were detected. We conclude that, although IGF-I concentrations are dependent on the phase of the menstrual cycle, the variation in IGF-I concentrations throughout the menstrual cycle is relatively small. Therefore, the menstrual cycle does not need to be considered when evaluating IGF I or IGFBP-3 serum values in women suspected to have GH deficiency. PMID- 9402267 TI - Utilization of retrieved oocytes as an index of the efficiency of superovulation strategies for in-vitro fertilization treatment. AB - An analysis of 581 in-vitro fertilization treatment cycles was carried out to determine the pattern of utilization of retrieved oocytes. Patients were divided into five groups depending on the number of retrieved oocytes. The mean fertilization rate of 57% was broadly similar amongst the groups but the proportion of retrieved oocytes that produced embryos of a quality suitable for transfer or cryopreservation (the cycle efficiency index) fell significantly with increase in the retrieved oocyte number. However, the pregnancy rate increased with the number of retrieved oocytes (13-38%). It is important to determine the point at which advantages of multifollicular development are outweighed by the potential for complications. Increased utilization of retrieved oocytes will decrease the need for production of a large number of oocytes. PMID- 9402268 TI - Recombinant human follicle stimulating hormone (r-hFSH; Gonal-F) versus highly purified urinary FSH (Metrodin HP): results of a randomized comparative study in women undergoing assisted reproductive techniques. AB - A prospective, randomized, comparative, assessor-blind study was carried out in two centres to compare the efficacy and safety of recombinant human follicle stimulating hormone (r-hFSH; Gonal-F) versus highly purified urinary FSH (u-hFSH HP; Metrodin HP), both administered s.c. in women undergoing ovarian stimulation for in-vitro fertilization including intracytoplasmic sperm injection (ICSI). A total of 235 patients started a long gonadotrophin-releasing hormone agonist protocol: 119 received r-hFSH and 114 received u-hFSH HP (150 IU/day) for the first 6 days. Two patients were excluded from the study because they mistakenly received the incorrect treatment combination. Human chorionic gonadotrophin (HCG; 10000 IU, s.c.) was administered once there was at least one follicle 18 mm in diameter and two others > or = 16 mm. In all, 119 (100%) and 102 (89%) of the patients respectively in the r-hFSH and u-hFSH HP groups achieved the criteria for HCG. The mean numbers (+/- SD) of oocytes recovered (the primary endpoint) were 12.2 +/- 5.5 and 7.6 +/- 4.4 in the r-hFSH and u-hFSH HP groups respectively (P < 0.0001). However, the number of FSH treatment days (11.0 +/- 1.6 versus 13.5 +/- 3.7) and the number of 75 IU ampoules (21.9 +/- 5.1 versus 31.9 +/- 13.4) used were significantly less (P < 0.0001) in the r-hFSH group than in the u-hFSH HP group. In patients treated using ICSI (63 patients in each group), no difference in oocyte maturation was observed. The mean numbers of embryos obtained were 8.1 +/- 4.2 and 4.7 +/- 3.5 (P < 0.0001), in favour of the r-hFSH group. In the majority of patients (96 and 99% respectively) only one or two embryos were replaced (mean 2.0 +/- 0.2 and 1.9 +/- 0.1 respectively) in the r hFSH and u-hFSH HP groups. The clinical pregnancy rates per started cycle and per embryo transfer were 45 and 36%, and 48 and 47%, respectively in the r-hFSH and u hFSH HP groups (not significant). There were six (5.1%) and two (1.7%) cases of ovarian hyperstimulation syndrome respectively. In conclusion, it was found that r-hFSH was more effective than u-hFSH at inducing multiple follicular development. However, the high rate of low ovarian response in the u-hFSH group compared with our general experience was unexpected. The availability of a gonadotrophin with less inter-batch variation would be beneficial for clinicians. r-hFSH seems to fulfil such a requirement. PMID- 9402269 TI - Beta2-glycoprotein I-dependent anticardiolipin antibody in early recurrent spontaneous abortion. AB - The objective of this study was to assess the clinical significance of autoimmune anticardiolipin antibody that can react with cardiolipin only in the presence of beta2-glycoprotein I (beta2-glycoprotein I-dependent anticardiolipin antibody) in the pathogenesis of early recurrent abortion. A total of 72 early recurrent spontaneous aborters and 175 normal healthy women were analysed for the occurrence of beta2-glycoprotein I-dependent anticardiolipin antibody in serum samples by an enzyme-linked immunosorbent assay specific for the detection of beta2-glycoprotein I-dependent anticardiolipin antibody. The incidence of beta2 glycoprotein I-dependent anticardiolipin antibody in the early recurrent spontaneous aborters was essentially the same as that of normal women. Thus, the beta2-glycoprotein I-dependent anticardiolipin antibody seemed to have little, if any, implication in the pathogenesis of early recurrent spontaneous abortion. PMID- 9402270 TI - Leukocytosis in response to exogenous gonadotrophin stimulation. AB - Leukocytosis may develop in women undergoing ovulation induction. The production of blood leukocytes and their numbers in circulation are regulated by complex interactions involving endogenous haematopoietic cytokines, such as granulocyte colony stimulating factor (G-CSF), monocyte-colony stimulating factor (M-CSF), and interleukins. The purpose of this prospective study was to explore the presence of leukocytosis in women who receive urinary menotrophins, and to determine whether haematopoietic cytokines are changed in the stimulation process. Controls were volunteers of the same age range, not taking any medication, who received daily saline injections. Subjects underwent phlebotomy at defined points for determination of complete blood counts, G-CSF, M-CSF, and interleukin-6 concentrations. Baseline white blood cell (WBC) counts were similar in patients and controls. In menotrophin-treated patients the WBC counts rose significantly (4.19 +/- 0.28 to 6.37 +/- 0.71) during the stimulation and peaked in the luteal phase (P = 0.037). In contrast, WBC counts decreased in controls. Other leukocytic lineages were not affected. In treated patients G-CSF concentrations rose significantly (P = 0.028 versus controls), while changes in M CSF and interleukin-6 were not significant. PMID- 9402271 TI - Is the outcome of in-vitro fertilization and embryo transfer treatment improved by spontaneous or surgical drainage of a hydrosalpinx? AB - A pilot study was designed to examine whether the outcome of embryo transfer in women with a hydrosalpinx might be improved by surgical drainage of the hydrosalpinx at the time of oocyte collection for in-vitro fertilization treatment. A comparative, controlled but retrospective analysis of the results was performed of all women with infective tubal damage aged <40 years old, who had ovulatory cycles, a normal uterus and a partner with normal spermatozoa. A standardized treatment regimen was used. A maximum of three embryos were transferred. Hydrosalpinx was defined by prior hysterosalpingography and/or laparoscopy with transcervical dye injection. A total of 237 embryo transfer cycles in women with hydrosalpinges (tubal distension not visible in 151, visible but not drained in 30 and drained in 56) were compared with 705 embryo transfer cycles in women with tubal disease but no hydrosalpinx. Results were analysed in the first three cycles but also separately in the first cycle to check for bias. Success rates were higher in the first cycle, but did not significantly influence overall differences. Implantation rates were significantly reduced overall in the hydrosalpinx group (8.0 versus 13.2% for controls; P < 0.001), being 8.3% (P < 0.01) in the subgroup without evident tubal distension and 7.5% (not significant) in the drained hydrosalpinx group. This study shows that tubal damage with distal occlusion is associated with a marked reduction in embryo implantation, even in the absence of obvious fluid distension. Surgical drainage of distended hydrosalpinges appears to offer no benefit. PMID- 9402272 TI - Sonohysterographic evaluation of uterine abnormalities noted on hysterosalpingography. AB - Transvaginal sonohysterography was performed on 40 consecutive patients with infertility or recurrent pregnancy loss and uterine abnormalities on hysterosalpingography. The findings were correlated with the hysterosalpingogram and subsequent diagnostic and/or operative hysteroscopy. Hysterosalpingography was incorrect in nine cases. Sonohysterography was more accurate than hysterosalpingography and provided more information about uterine abnormalities. Sonohysterography was in complete agreement with hysteroscopy. Diagnostic hysteroscopy can therefore be avoided if the sonohysterogram is normal. Sonohysterography also provides additional information on the relative proportion of the intracavitary and intramyometrial components of submucus myomas, as well as extracavitary myomas and the adnexae. This may help in planning the surgical procedure. PMID- 9402273 TI - Analyses of meiotic chromosomes in testicular biopsies of infertile patients. AB - Between 1989 and 1992 meiotic chromosome studies and synaptonemal complex analyses were evaluated using light and, in part, electron microscopy in 46 infertile males with highly abnormal spermiograms. This examination focused on whether the breakdown of spermatogenesis could be attributed to pairing anomalies of bivalents. The study of meiotic chromosomes and synaptonemal complexes indicated normal spermatogenesis in five patients (11%); in the remainder, maturation arrest was diagnosed. In 21 individuals (50%) the breakdown was accompanied by pairing anomalies (asynapsis, fragmented synaptonemal complexes, X/Y univalence). Thus it is shown that male infertility can often partly be explained by meiotic disorders. PMID- 9402274 TI - Treatment for infertility and risk of invasive epithelial ovarian cancer. AB - The relationship between the use of fertility drugs and the risk of ovarian cancer was analysed using data from an Italian case-control study. The study comprised 971 women below the age of 75 years with histologically confirmed invasive epithelial ovarian cancer diagnosed within the year before the interview. The controls were 2758 women admitted to the same network of hospitals where the cases of ovarian cancer had been identified. Five cases (0.5%) and 11 controls (0.4%) reported use of fertility drugs. In comparison with women who had never used fertility drugs, the multivariate odds ratio (OR) for women who had taken fertility drugs was 1.1 [95% confidence interval (CI) 0.4-3.3]. The OR were 0.7 (95% CI 0.1-7.9) and 1.0 (95% CI 0.2-3.8) for women who had used fertility drugs for <6 and > or =6 cycles respectively. Considering the 14 cases and 45 controls reporting difficulty in conception, the risk of ovarian cancer was 0.5 (95% CI 0.1-3.6) for women who reported use of fertility drugs. Considering nulliparous women only, the estimated OR of ovarian cancer for any fertility drug use was 0.6 (95% CI 0.1-3.5). Although the present results have limitations in terms of statistical power and available information, they provide reassuring evidence of the absence of a strong association between fertility drugs and subsequent risk of developing epithelial ovarian cancer. PMID- 9402275 TI - Intrauterine insemination: effect of the temporal relationship between the luteinizing hormone surge, human chorionic gonadotrophin administration and insemination on pregnancy rates. AB - The optimal time period for intrauterine insemination (IUI) in relation to either luteinizing hormone (LH) surge or human chorionic gonadotrophin (HCG) administration leading to the best pregnancy rates has not been determined. In this study, 856 consecutive human menopausal gonadotrophin (HMG)-stimulated and 49 natural unstimulated IUI cycles carried out at a reproductive medicine unit affiliated with a tertiary centre were analysed in a retrospective fashion. There were three scenarios in the temporal relationship of the LH surge, HCG administration and artificial insemination. These were (group A) subjects who had an endogenous LH surge but were not given HCG; (group B) subjects who were given HCG after an observed LH surge, and (group C) subjects who were given HCG before the LH surge. The overall pregnancy rate (PR) was 16% per cycle. The PR was 9% in group A, 20% in group B and 14% in group C. The PR in group B was significantly better than group C (P = 0.04). In group B, the longer the time interval between the LH surge and HCG administration, the better the PR up to 20 h (P = 0.025); the timing of IUI based on the LH surge was not critical to the achievement of pregnancy within 3 days. In group C, PR improved with the increasing interval between HCG and IUI from <28 h up to 60 h. We conclude that a better PR is achieved if a spontaneous LH surge occurs before HCG administration, especially where the administration of HCG is delayed 8-20 h after an observed LH surge; the timing of IUI based on the LH surge is not critical to the achievement of pregnancy within 3 days. PMID- 9402276 TI - Thromboembolic disease associated with ovarian stimulation and assisted conception techniques. AB - Thromboembolic disease, as a complication of ovarian stimulation and assisted conception techniques, is generally considered to be a rare complication of ovarian hyperstimulation syndrome and, by implication, lower limb in origin. Sporadic cases of unusually sited thromboses, both venous and arterial, have been reported. This paper aims to draw attention to the relatively large number of such thromboses reported in the world literature compared with those cited in previous commentaries, and to emphasize how little is known about their pathogenesis. It is believed that this is an issue which requires to be addressed in order to understand the background pathology to such incidents and if possible to identify women at greatest risk from such potentially debilitating or fatal complications, such that appropriate prophylactic measures can be taken. PMID- 9402277 TI - Upper limb thrombosis associated with assisted conception treatment. AB - Three cases of upper limb deep venous thrombosis occurring in association with assisted conception treatment are presented. The accepted argument that lower limb thrombosis occurring in cases of complicated or severe hyperstimulation syndrome represents the likeliest thrombo-embolic disorder in this situation is questioned. PMID- 9402278 TI - Hamster origin of metaphases with multiple chromosome rearrangements in first cleavage human-hamster embryos. AB - Laboratories using the human sperm-hamster egg fertilization system to analyse sperm chromosomes obtain, sporadically, metaphases with multiple aberrations. Due to the high number of aberrations, these metaphases cannot be fully karyotyped. In some of them, one or several human chromosomes can be identified, guaranteeing the human origin of the whole metaphase. However, in others, none of the chromosomes can be recognized as human. This latter type of grossly rearranged metaphases is characterized by complex chromatid exchanges, multifragmented chromosomes and pulverized chromosome material. Using fluorescent in-situ hybridization techniques (FISH) with either human or hamster genomic DNA probes, we examined the origin of this second type of metaphase with multiple chromatid exchanges and fragmented chromosomes. Our study demonstrates that all of them hybridize with hamster genomic DNA probes and not with human DNA, proving their hamster origin. Since some of these metaphases seem to be diploid, we suggest that they may arise from hamster eggs that have failed to complete meiosis and have not extruded the second polar body. PMID- 9402279 TI - No evidence for a decreased fertilizing potential after in-vitro fertilization using spermatozoa from polyzoospermic men. AB - Polyzoospermia is generally recognized as a male factor contributing to infertility and/or recurrent abortion. Although a reduced spermatozoal fertilizing capacity is assumed to be involved, so far there is no conclusive explanation for the assumed reduced reproductive performance in these patients, and data on the fertilizing capacity of spermatozoa from polyzoospermic men are lacking. The present study therefore aimed at analysing the outcome after in vitro fertilization (IVF)-embryo transfer in polyzoospermic patients. Retrospective analysis showed that only 0.5% out of 7863 IVF cycles were performed with spermatozoa from polyzoospermic men. The outcome of these IVF cycles shows neither a reduction in spermatozoal fertilizing capacity nor an increase in pregnancy wastage in cycles in which a pregnancy was obtained. These results may suggest a normal reproductive potential in polyzoospermic patients and therefore the question may be raised whether polyzoospermia represents a real pathological entity leading to infertility. PMID- 9402280 TI - Is transrectal ultrasonography a reliable diagnostic approach in ejaculatory duct sub-obstruction? AB - We studied the diagnostic predictive power of transrectal ultrasonography (TRUS) coupled with semen volume in cases of distal seminal tract sub-obstruction. As a gold standard for diagnosis we used seminal tract washout (STW). Non-azoospermic subjects (n = 112) were submitted to transrectal ultrasonography because of suspected excretory infertility or other andrological pathologies, before performing STW. STW indicated ejaculatory duct sub-obstruction in 36.6% of the patients. Seminal vesicle enlargement (anterior-posterior diameter > or = 15 mm) and seminal vesicle roundish anechoic areas (stasis) were the ultrasonographic anomalies more often associated with ejaculatory duct sub-obstruction. Stepwise logistic regression (SLR) analysis revealed that the ultrasonographic evidence of stasis was highly diagnostic only in the presence of a low semen volume (< or = 1.5 ml) and that ejaculatory duct sub-obstructions may be present but with no evidence of ultrasonographic anomalies. Therefore, TRUS is a useful approach for the treatment of suspected ejaculatory duct sub-obstruction, but is not a reliable diagnostic tool and, before performing transurethral surgery, STW should be mandatory. PMID- 9402282 TI - Immunomodulation by human seminal plasma: a benefit for spermatozoon and pathogen? AB - The immunosuppressive effect of human seminal plasma and its implications for sperm survival are reviewed. Human semen contains high concentrations of prostaglandins that can effect a cytokine-mediated switch away from a cell mediated immune response. This effect on antigen presenting cells would induce a state of non-responsiveness to sperm antigens in the female reproductive tract. It is postulated that the induction of anergy to sperm antigen may be fundamental to the continuing fecundity of the individual. However, although this immune system modulation will benefit the spermatozoa, the response to infective agents present in semen will also be affected, which may play a critical role in the aetiology and progress of sexually transmitted disease. PMID- 9402281 TI - Pentoxifylline induces hyperactivation and acrosome reaction in spermatozoa of golden hamsters: changes in motility kinematics. AB - Pentoxifylline (PF) is used to enhance motility of spermatozoa from infertile human subjects. We have previously shown that 0.45 mM PF improved capacitation of spermatozoa and fertilization of oocytes in vitro in hamsters. The present study was carried out to assess PF-induced changes in motility kinematics of hamster spermatozoa by a computer-aided sperm analyser (CASA) and determine the timing of onset of hyperactivation (HA) and acrosome reaction (AR) in PF-treated spermatozoa. Motility kinematics were analysed by CASA for 0-8 h in the absence or presence of 0.45 mM PF in Tyrode's medium supplemented with lactate, pyruvate and polyvinyl alcohol (TLP-PVA) or in TLP-PVA with bovine serum albumin (TALP PVA). Conventional assessment was also made on the percentage of motility and quality of motility of spermatozoa; values were expressed as sperm motility index (SMI). Both in TALP-PVA and TLP-PVA, PF markedly increased SMI, especially the quality of motility (P < 0.02) by 2-3 h which was sustained up to 6 h. The motility kinematic data of PF-treated spermatozoa in TALP-PVA showed that average path velocity, curvilinear velocity and amplitude of lateral head displacement significantly (P < 0.05) increased as early as 2 h, with the expected decrease in straightness (STR) and linearity (LIN). Similar changes were also observed with PF-treated spermatozoa in TLP-PVA. Moreover, the percentage of hyperactivated spermatozoa in PF-treated samples was significantly (P < 0.001) higher than the untreated control at 2 h. To determine whether PF could induce AR, independent of bovine serum albumin, quantitative AR was assessed by observing the presence or absence of acrosomal cap on viable spermatozoa. PF significantly (P < 0.001) increased the percentage of AR as early as 2 h, reaching maximum at 4 h both in TALP-PVA (P < 0.05) and in TLP-PVA (P < 0.001). These results show that, in hamsters, PF induces early onset (by 2 h) of HA and AR and increases the proportion of spermatozoa undergoing physiological maturation. PMID- 9402283 TI - A simple and objective approach to identifying human round spermatids. AB - Although round spermatids have been studied extensively using staining techniques and electron microscopy, little information is available about their appearance in living conditions. We describe a method of collecting and identifying round spermatids from ejaculates and testicular biopsies. The validity of the selection procedure was confirmed by fluorescence in-situ hybridization. Based on cell size, morphological characteristics of nucleus and cytoplasm, and on the nucleus/cytoplasm ratio, we harvested a population of cells that was 84% haploid. This procedure can be applied to select spermatids for clinical or research purposes. PMID- 9402284 TI - Microsurgical epididymal sperm aspiration with motile trophozoite cells but no spermatozoa. AB - This paper reports on a patient in whom the clinical diagnosis of obstructive azoospermia was made according to clinical observations, i.e. azoospermia, normal andrological examination, normal follicle stimulating hormone and a misleading histopathological report of a testicular biopsy. Microsurgical vasoepididymostomy failed to restore fertility, and as a last resort, microsurgical sperm aspiration was performed. Although flagellated cells were observed in the epididymal aspiration, no spermatozoa were observed and wet preparation of multiple testicular biopsies failed to demonstrate any spermatozoon. This patient was diagnosed to have a non-obstructive azoospermia, resulting from maturation arrest associated with trichomonas infection at the level of the epididymis. PMID- 9402285 TI - Possible contribution of follicular interleukin-1beta to nitric oxide generation in human pre-ovulatory follicles. AB - The aim of this study was to investigate the relationships between follicular nitric oxide (NO) metabolite concentrations and several related variables, with special reference to follicular interleukin-1beta (IL-1beta). The follicular fluid from the leading and secondary follicles was collected individually from 20 women undergoing in-vitro fertilization (IVF) treatment, and the concentrations of nitrite (NO2-) and nitrate (NO3-) were determined fluorometrically using 2,3 diaminonaphthalene. Both follicular nitrite (r = 0.42, P < 0.01) and nitrate (r = 0.49, P < 0.001) were found to be significantly correlated with follicular IL 1beta concentrations. There were also significant positive correlations between follicular nitrate and the number of oocytes retrieved (P < 0.01) and serum oestradiol concentration on the day of human chorionic gonadotrophin (HCG) administration (P < 0.05). When follicular cells were incubated in vitro with 10 ng/ml of IL-1beta for 24 h, nitrate generation was significantly (P < 0.01) elevated compared with the control. In conclusion, our study demonstrates that follicular IL-1beta and the number of developing follicles are significant variables that affect follicular NO concentrations, and points to the possible contribution of IL-1beta to NO generation in human preovulatory follicles. PMID- 9402286 TI - A novel mechanism of vascular endothelial growth factor, leptin and transforming growth factor-beta2 sequestration in a subpopulation of human ovarian follicle cells. AB - This study describes the occurrence of a highly specialized subpopulation of granulosa and cumulus oophorus cells that accumulate and sequester specific growth factors by a novel mechanism. These cells are characterized by multiple balloon-like processes tethered to the cell by means of a slender stalk of plasma membrane. Time-lapse analyses demonstrate that these tethered structures (TS) form in minutes and frequently detach from the cell with the bulbous portion remaining motile on the cell surface. Serial section reconstruction of transmission electron microscopic images shows a specific and stable intracellular organization in which an apparent secretory compartment composed of densely packed vacuoles, vesicles, and cisternae is separated by a thick filamentous network from a nuclear compartment containing mitochondria, polyribosomes, lipid inclusions, and rough-surfaced endoplasmic reticulum. Immunofluorescent analysis performed during the formation of these structures showed a progressive accumulation of vascular endothelial growth factor, leptin, and transforming growth factor-beta2 in the bulbous region. TS were identified in newly aspirated masses of granulosa and cumulus oophorus, and their production persists for months in culture. Observations of TS-forming cells made over several days of culture indicates that their production is episodic and factor release from these cells may be pulsatile. The findings suggest that a novel method of growth factor storage and release by an apparent apocrine-like mechanism occurs in the human ovarian follicle. The results are discussed with respect to possible roles in pre- and post-ovulatory follicular development. PMID- 9402287 TI - Apoptosis of germ cells during human prenatal oogenesis. AB - During human oogenesis two contrasting processes can be observed: germ cell proliferation and differentiation, and contemporaneous germ cell death. It is well known that apoptosis is a type of physiological cell death that occurs in proliferating and differentiating tissues. The aim of this study is to demonstrate, through ultrastructural and in-situ 3' end labelling observations in intact sections of human fetal ovaries, that germ cell loss during fetal life is due to a phenomenon of apoptosis. We evaluated the presence of programmed cell death in female germ cells in fetal ovaries at 18-20 weeks of postconceptional age. The alterations that occur during apoptosis were detected by the electron microscope and include cytoplasmic condensation, organelle relocalization and compaction, chromatin condensation, and finally, production of membrane-enclosed particles containing intracellular material, known as apoptotic bodies, that are phagocytosed. The fragmentation of DNA, characteristic of apoptotic cells, was detected by the use of the in-situ 3' end labelling procedure on histological sections of ovaries where only some nuclei of germ cells were positively stained. The parallel use of these two methods on human ovaries of 18-20 weeks postconceptional age has allowed us to show that the numerical decrease of human female germ cells during the fetal period is due to an apoptotic phenomenon. PMID- 9402288 TI - Ultrastructural observations in human oocytes and preimplantation embryos after zona opening using an erbium-yttrium-aluminium-garnet (Er:YAG) laser. AB - For more than 3 years we have performed laser-assisted hatching prior to embryo transfer in patients with recurrent implantation failure using an erbium-yttrium aluminium-garnet (Er:YAG) laser system that operates in the infrared region of the light spectrum. The laser beam is guided through a quartz fibre and is brought into direct contact with the zona pellucida. This study was undertaken to evaluate the ultrastructural effects of this laser on the zona pellucida and underlying cell membrane of unfertilized human oocytes and pathologically fertilized preimplantation embryos using light and scanning electron microscopy. The Er:YAG laser produces an almost circular zona opening in the shape of a truncated cone tapering off towards the inside, with a mean diameter of 18 mm. The exact diameter of the drilled site depends on the diameter of the fibre tip and the total number of pulses applied. After laser interaction, the zona matrix and the surface of the underlying ooplasm membrane showed no degenerative alterations. We conclude that the Er:YAG laser is an effective microsurgical tool for achieving reproducible, precise zona openings particularly suitable for purposes of assisted hatching because of their characteristic shape. PMID- 9402289 TI - Effects of different hyaluronidase concentrations and mechanical procedures for cumulus cell removal on the outcome of intracytoplasmic sperm injection. AB - The aim of this study was to compare concentrations of hyaluronidase and mechanical methods used to denude human oocytes from surrounding cumulus and corona cells prior to intracytoplasmic sperm injection (ICSI). Cumulus and corona cells were removed in two pipetting steps: first in a medium containing hyaluronidase, and then in a medium without enzyme. The first step in the procedure was investigated. Different hyaluronidase concentrations (78, 39 or 10 IU/ml) and pipettes of different size (inner diameter 250 or > 1000 microm) were used to remove the cumulus cells. The time required to denude the oocytes was recorded. Metaphase II oocytes were injected, and the survival, fertilization, embryo development and pregnancy rates were evaluated. We found that by using a pipette with an inner diameter of at least 1000 microm we were able to decrease significantly the time an oocyte is exposed to hyaluronidase, even if the concentration of enzyme is very low (10 IU/ml). For the different conditions there was no statistically significant effect on the outcome in terms of survival, normal fertilization [two pronuclear (2PN)], parthenogenetic activation (1PN), abnormal fertilization (3PN), embryo development and pregnancy rates after ICSI. In conclusion, a concentration as low as 10 IU/ml hyaluronidase in combination with a pipette of at least 1000 microm inner diameter can be used successfully to denude oocytes for microinjection. PMID- 9402290 TI - Transcription of tissue-specific genes in human preimplantation embryos. AB - We have recently detected de-novo transcripts of the predominantly muscle specific myotonin protein kinase gene in human preimplantation embryos from the 1 cell to the 4-cell stages. Others have shown de-novo transcripts of the Y-linked genes, ZFY and SRY, in the 1-cell zygote. In order to assess the significance of early transcription of these predominantly tissue-specific genes in preimplantation development, we have analysed individual human oocytes and preimplantation embryos for the presence of transcripts of two further tissue specific genes, alpha-globin and beta-globin, and two house-keeping genes, HPRT and APRT. Reverse transcriptase polymerase chain reaction assays were developed to the required single cell sensitivity, using human red blood cells and fibroblasts, prior to their application to human oocytes and embryos. As expected, transcripts of the house-keeping genes, HPRT and APRT, were detected at all stages of preimplantation development. Transcripts of 'tissue-specific' alpha globin were readily detected in preimplantation embryos from the 1-cell stage. However, transcripts of beta-globin were detected only rarely (in only one of the 11 embryos analysed). This difference may be due to the fact that alpha-globin contains a CpG island. A survey of the data on gene expression in early human development suggests that CpG-island-containing genes may be expressed in preimplantation embryos. Expression of these genes in gametes and early embryos may be involved in the survival of CpG islands in evolution. PMID- 9402291 TI - The chromosomal constitution of multipronuclear zygotes resulting from in-vitro fertilization. AB - We have attempted to analyse the chromosome constitution of 77 multipronuclear uncleaved zygotes obtained from our in-vitro fertilization programme. Complete karyotypes could be established for 51 tripronuclear cells and eight zygotes with four pronuclei. When compiling the results, the varying arrangement of the chromosome sets was taken into consideration. Eighteen tripronuclear zygotes showed three separate haploid metaphases (distribution pattern n/n/n), 16 cells had one haploid and one diploid chromosome set (n/2n), and in 15 zygotes the individual sets were not distinguishable (3n). Two zygotes were in fact tetraploid, the distribution of metaphases on the slide being n/3n and n/n/2n, respectively. In tripronuclear zygotes the sex chromosome ratio XXX:XXY:XYY was 14:16:18, excluding the two tetraploid cells and one zygote with a 23,X/23,X/22, C or -Y karyotype. Chromosome abnormalities were found in 16 zygotes (31.4%) and included numerical (six cells), structural (four cells) as well as combinations of numerical and structural alterations (six cells). Four of the zygotes with four pronuclei (50%) had numerical and/or structural chromosome aberrations. Excluding two cells with one uninterpretable metaphase and a 22,-C or -Y karyotype, respectively, the sex chromosome distribution XXXX:XXXY:XXYY:XYYY was 1:1:2:1 in zygotes with four pronuclei. Another zygote was found to be pentaploid after fixation. These results suggest that analysis of multipronuclear zygotes yields valuable information about cytogenetic abnormalities occurring at the earliest stage of conception. PMID- 9402292 TI - Prospective randomized study comparing human serum albumin with fetal cord serum as protein supplement in culture medium for in-vitro fertilization. AB - The use of human serum albumin (HSA) instead of fetal cord serum (FCoS) as protein supplement highly simplifies the preparation of culture medium for human in-vitro fertilization (IVF) but whether they are equivalent in sustaining embryo development is still controversial. We performed a prospective randomized study of patients undergoing IVF or intracytoplasmic sperm injection (ICSI) where embryos were cultured in Earle's balanced salt solution containing either 8% (v/v) FCoS or 0.4% (w/v) HSA as protein source. Fertilization rates, morphological embryonic quality and pregnancy rates were compared. A total of 2189 oocytes from 210 cycles were cultured in medium supplemented with HSA in patient group 1 and 2109 oocytes from 203 cycles in medium supplemented with FCoS in patient group 2. The fertilization rate, defined as the presence of two nuclei, for microinjected oocytes was similar in both patient groups (77.4 and 76.7%, respectively). The fertilization rate for inseminated oocyte-cumulus complexes was significantly higher in the HSA group than in the FCoS group (62.9 versus 53.8%, P < 0.025). The embryonic quality was significantly better after culture in medium supplemented with HSA than with FCoS (13.7 versus 9.9% morphologically excellent embryos, P < 0.001). Implantation rates per transferred embryo were not significantly different (22.5 versus 18.2%), but there was a significantly higher pregnancy rate per embryo transfer in the HSA group (45.7 versus 35.9%, P < 0.05, respectively). Non-evolutive pregnancy rates were significantly different (27.4 and 16.7%). Our data demonstrate that the use of human serum albumin as a protein supplement for culture medium in human IVF programmes is associated with improved embryonic quality and significantly higher pregnancy rates. For this reason as well as the additional benefits of being virus-free and being purified, HSA is preferable to FCoS for the preparation of culture media in human IVF. PMID- 9402293 TI - Induction of artificial endometrial cycles with s.c. oestrogen implants and injectable progesterone in in-vitro fertilization treatment with donated oocytes: a preliminary report. AB - Endometrial preparation for embryo implantation in oocyte recipients with retained ovarian function presents a special problem. In all, 10 women with preserved ovarian function received donated oocytes in an in-vitro fertilization programme. In the preparatory cycles with oral oestrogens, all failed to develop adequate secretory endometrium because of ill-timed early luteinization occurring during the proliferative phase of the cycle. Subsequently the patients were treated with s.c. 17-beta oestradiol implants and injectable progesterone. The implants successfully induced complete down-regulation of the hypothalamus, pituitary axis, and prevented luteinizing hormone (LH) surge. In these preparatory cycles, all the treated patients produced adequate secretory endometrium. Clinical trials of 27 oocyte donation cycles yielded one biochemical and eight clinical pregnancies. PMID- 9402294 TI - Endometrial thickness and serum oestradiol concentrations as predictors of outcome in oocyte donation. AB - Adequate endometrial preparation with exogenous steroids is mandatory for successful ovum donation. This study was undertaken to assess the value of endometrial thickness by ultrasound and serum oestradiol as predictors of ovum donation outcome and to analyse the correlation between serum oestradiol concentrations and the endometrial thickness. Endometrial thickness and serum oestradiol concentrations on the day of oocyte donation were recorded and compared to several in-vitro fertilization parameters. The cycles (n = 465) were classified according to serum oestradiol values and endometrial thickness. Comparison of the groups showed that endometrial thickness was significantly (P = 0.002) higher when serum oestradiol was >400 pg/ml as compared to <100 pg/ml. Pregnancy and implantation rates did not differ among the groups, women with serum oestradiol <50 pg/ml having similar outcome to the remaining cases. Endometrial thickness showed a similar picture in terms of pregnancy and implantation. Also, women with an endometrium <4 mm in size had normal pregnancy and implantation rates. There was a positive correlation (P = 0.0044) between endometrial thickness and implantation, as well as between endometrial thickness and serum oestradiol (P = 0.0184). None of the parameters examined was able to predict ovum donation outcome. It is concluded that endometrial thickness is preferred to serum oestradiol for the monitoring of endometrial development, although neither is able to predict success in oocyte donation. PMID- 9402295 TI - Reproductive impact of congenital Mullerian anomalies. AB - This retrospective longitudinal study was undertaken in order to determine the incidence and reproductive impact of uterine malformations on women desiring to conceive during their reproductive years. A total of 3181 patients in whom the morphology of the uterus was ascertained by hysterosalpingography (HSG) and laparoscopy/laparotomy during the years 1980-1995 was included in the study. The population analysed included fertile, infertile and sterile patients. The overall frequency of uterine malformations was 4.0%. Infertile patients (6.3%) had a significantly (P < 0.05) higher incidence of Mullerian anomalies, in comparison with fertile (3.8%) and sterile (2.4%) women. Septate (33.6%) and arcuate (32.8%) uteri were the most common malformations observed. Each malformation was individually analysed in fertile and infertile patients, in order to ascertain its actual reproductive impact. The performance of the unicornuate and didelphys uteri was similar with a chance of having a living child of 37-40%. The reproductive potential of the bicornuate uterus showed a live birth rate of 62.5% and the septate uterus showed a live birth rate of 62%. In all these abnormalities, early miscarriages (25-38%) and preterm deliveries (25-47%) were quite common. The arcuate uterus presented a live birth rate of 82.7%. It is concluded that uterine anomalies are relatively frequent in fertile women, and more frequent in infertile patients. Nevertheless, fertile patients with normal reproductive performance do exist, and Mullerian defects can permit an absolutely normal obstetric outcome. The reproductive performance of the unicornuate and didelphys uteri was poor, while that of the septate and bicornuate uteri was better than expected. The arcuate uterus had no impact on reproduction. PMID- 9402296 TI - The impact of endometriosis in couples undergoing intracytoplasmic sperm injection because of male infertility. AB - To assess the impact of endometriosis on intracytoplasmic sperm injection (ICSI) outcome, we have retrospectively evaluated 980 ICSI cycles, comparing the results of women with and without endometriosis. A total of 101 cycles was identified in which various degrees of endometriosis were involved, and in the remaining 879 cycles, male infertility was the only cause of infertility. Ejaculated spermatozoa were microinjected in all cycles. There was a significant reduction (P = 0.004) in the number of oocytes retrieved from women with endometriosis as compared to those without endometriosis. However, there were no significant differences in either fertilization or pregnancy and implantation rates between women with or without endometriosis. We conclude that the presence of endometriosis in patients undergoing ICSI because of severe male infertility does not affect fertilization, pregnancy and implantation rates, although significantly fewer oocytes are retrieved from patients with endometriosis. PMID- 9402297 TI - Intrauterine sonographic assessments of embryonic heart diameter. AB - Our purpose was to evaluate embryonic heart diameter in early first-trimester pregnancy using intrauterine sonography with a 20 MHz flexible catheter-based high-resolution real-time miniature transducer. A total of 40 women about to undergo therapeutic abortion from 6-9.9 weeks gestational age and one abnormal pregnancy with fetal hydrops at 9 weeks were studied with a specially developed catheter-based high-resolution real-time miniature (2.4 mm outer diameter) ultrasound transducer (20 MHz). A curvilinear relationship was found between the menstrual age and embryonic heart diameter (R2 = 95.7%), and a normal range of embryonic heart diameter for estimating the growth of the embryonic heart during early first-trimester pregnancy was generated. A normogram of menstrual age as predicted by embryonic heart diameter was also established. There was a good curvilinear correlation between embryonic heart diameter and crown-rump length (R2 = 90.1%). The embryonic heart diameter/crown-rump length ratio rapidly decreased from week 6 to week 7, and remained almost constant thereafter. Embryonic heart diameter (5.2 mm) in the case of fetal hydrops at 9 weeks was above the normal range. These results may provide an additional method of estimating gestational age in the early first trimester of pregnancy. In this limited series, a single case of embryonic heart enlargement was demonstrated, suggesting its potential use in the detection of embryonic congestive heart failure. PMID- 9402298 TI - Serum levels of folate and cobalamin in women with recurrent spontaneous abortion. AB - We evaluated the folate and cobalamin status in 29 non-pregnant women with a history of recurrent spontaneous abortion (three or more consecutive) of unknown aetiology in comparison to 29 healthy nulligravidae of similar reproductive age (controls). Serum concentrations of folate and cobalamin showed no significant differences between the two groups. No correlation between age and vitamin concentrations was found. In the study group there was a significant negative correlation of the number of previous abortions and the concentration of serum folate. Patients with at least four previous miscarriages had significantly lower serum values of folic acid than women with three abortions, but not than controls. The underlying cause of this finding remains unclear. In conclusion, the serum concentrations of folic acid and vitamin B12 are not significantly altered in women with unexplained recurrent spontaneous abortions, and an association between a deficiency of these vitamins and an increased risk of pregnancy loss appears to be questionable in the majority of gestations. PMID- 9402299 TI - Remodelling of guinea-pig aorta during pregnancy: selective alteration of endothelial cells. AB - It is known that aortic blood flow is increased during pregnancy, which may be due to a pregnancy-associated decrease in aorta sensitivity to vasoconstrictors on one side, and increased response to vasodilators on the other. Recent studies have shown that alteration of blood flow or pressure could remodel some arteries over a short time frame. However, the possibility of remodelling of aorta during pregnancy has not yet been examined. Therefore, the aim of the present study was to assess the morphometric and stereological characteristics of guinea-pig aorta during different stages of pregnancy (non-pregnant, early-pregnant, mid-pregnant, late-pregnant, n = 8-10 for each group). The cross-sectional areas of different aortic layers and of endothelial cells were measured using both light and electron microscopy. The values of external and internal diameters, wall thickness, total cross-sectional area and cross-sectional area of media and adventitia were not significantly different, regardless of the stage of pregnancy. In contrast, the cross-sectional area of intima significantly and progressively decreased during pregnancy (non-pregnant: 61 +/- 5 x 10(4) microm2, late-pregnant: 38 +/- 3 microm2, P < 0.01). The volume:surface density ratio of intima also significantly and progressively decreased during pregnancy (non pregnant: 5.31 +/- 0.51, late-pregnant: 4.38 +/- 0.42, P < 0.01). Electron microscopy revealed that the cross-sectional area of endothelial cells was significantly decreased during different stages of pregnancy (non-pregnant: 56.8 +/- 6.2 microm2, late-pregnant: 28.9 +/- 3.8 microm2, P < 0.01). It is concluded that during pregnancy there is selective thinning of intimal layer of guinea-pig aorta, which probably reflects hypotrophy of aortic endothelial cells. PMID- 9402300 TI - The obstetric implications of teenage pregnancy. AB - A retrospective review was performed on the obstetric outcome of teenage pregnancies delivered in 1 year in a tertiary centre. The results were compared with the rest of the obstetric population in the same hospital in the same year. The teenage mothers (n = 194) had increased incidence of sexually transmitted diseases (5.2 versus 1.0%, P < 0.05), and preterm labour (13.0 versus 7.0%, P < 0.01), but decreased incidence of gestational glucose intolerance (3.1 versus 11.4%, P < 0.001), when compared with the non-teenage mothers (n = 4914). There was no difference in the types of labour, while the incidence of Caesarean section was lower (4.1 versus 12.6%, P < 0.001) in the teenage mothers. Although the incidence of low birthweight was higher in the teenage mothers (13.5 versus 6.5%, P < 0.001), there was no significant difference in the mean birthweight, gestation at delivery, incidence of total preterm delivery, or perinatal mortality or morbidity. The results indicate that the major risk associated with teenage pregnancies is preterm labour, but the perinatal outcome is favourable. The good results accomplished in our centre could be attributed to the free and readily available prenatal care and the quality of support from the family or welfare agencies that are involved with the care of teenage mothers. PMID- 9402301 TI - Expression of superoxide dismutase and xanthine oxidase in myometrium, fetal membranes and placenta during normal human pregnancy and parturition. AB - Superoxide, an agent which attenuates the half-life of nitric oxide, is metabolized and synthesized by superoxide dismutase (SOD) and xanthine oxidase, respectively. Over the last few years much work has focused on the role of nitric oxide in human parturition. The aim of this study was to determine whether the onset of human parturition is associated with a change in the expression of copper/zinc superoxide dismutase (Cu/Zn SOD), manganese superoxide dismutase (Mn SOD) or xanthine oxidase within the uterus. Samples of myometrium, placenta, decidua and fetal membranes were obtained from women before and after the onset of labour at term. Immunocytochemistry was used to localize Cu/Zn SOD, Mn SOD and xanthine oxidase and measure SOD enzyme activity. Cu/Zn and Mn SOD-like immunoreactivity was detected in syncytiotrophoblast cells, villous stromal cells and endothelial cells of blood vessels in the placenta. In the myometrium Cu/Zn and Mn SOD were localized to myocytes and endothelial cells and to some vascular smooth muscle cells. In the fetal membranes we observed staining for Cu/Zn SOD and Mn SOD in the amnion, chorion, extravillous trophoblast and decidua. There was no difference in SOD enzyme activity or staining intensity for SOD between different cell types before and during labour. Xanthine oxidase immunoreactivity was identified in each of the tissues examined and again there was no difference in immunostaining in tissues obtained from women delivered before or after the onset of labour. These results show that the pregnant uterus is capable of both synthesizing and degrading superoxide and suggest that superoxide dismutase and xanthine oxidase may play a role in the maintenance of uterine quiescence during pregnancy, but not in the initiation of parturition. PMID- 9402303 TI - A shift from a male to a female majority in newborns with the increasing age of grand grand multiparous women. AB - In this longitudinal study, we investigated the relationship of birth order and the age of mother and father to the gender of 1795 newborns (mean +/- SD 12.5 +/- 1.6 per mother) of 143 grand grand multiparous (i.e women who have had >10 deliveries). The frequency of boys was 52.2% in the group of 1st to 9th paras and 46.2% in the group of 10th to 20th paras (P = 0.022). Mothers aged > or =35 years had 7.0% more female than male newborns (P = 0.024). The respective figure for fathers was 5.6% (P = 0.023). The interpregnancy interval evaluated for 96 mothers with 1091 deliveries had no correlation with the gender of the infants. In the stepwise logistic regression analysis, the age of the mothers remained the only significant independent factor for the shift from a male to a female majority in the newborns (P = 0.0389). The present data thus indicate that the age of the mother is the factor which explains why grand grand multiparous women deliver more girls than boys. PMID- 9402302 TI - Cyclooxygenase-1 and -2 in human placenta and placental bed after normal and pre eclamptic pregnancies. AB - In pre-eclampsia, the ratio of prostacyclin:thromboxane production rate is decreased favouring the vasoconstrictive thromboxane. One of the rate-limiting steps in prostaglandin synthesis is cyclooxygenase (COX) activity. Therefore, we investigated the expression of COX-1 and COX-2 in human placenta and placental bed. Tissue specimens from the 29th to 40th week of pregnancy were obtained from Caesarean sections after uncomplicated and pre-eclamptic pregnancies before the onset of labour. COX-1 and COX-2 were localized immunohistochemically with the identification of positive cells by double immunofluorescence staining. The protein and mRNA levels were analysed by immunoblotting and quantitative reverse transcriptase-polymerase chain reaction. Expression of both COX-1 and COX-2 could be observed in placenta and placental bed. COX-1-like immunoreactivity was observed in most cell types with strongest staining in macrophages. Only macrophages, endothelium, vascular leiomyocytes and fibroblasts stained positively for COX-2. In placenta, COX-1 and -2 expression was unchanged after pre-eclampsia. In placental bed, protein and mRNA levels of COX-1 were increased in the pre-eclamptic group (P < 0.05), whereas COX-2 expression did not differ significantly from normal pregnancies. An increased expression of COX-1 could be involved in the pathophysiology of pre-eclamptic changes within the placental bed. A therapy with drugs inhibiting COX-1 might be beneficial in this condition. PMID- 9402304 TI - A prospective study of psychosocial stress and fertility in women. AB - The objective of this study was to compare average stress levels during the month of conception to those of previous infertile months. We postulated that stress level during the actual month of conception would be lower than that during previous non-conception cycles. Thirteen normal women from the general community, who were attempting pregnancy, kept daily records of coital activity and basal body temperature, and twice a month completed self-administered questionnaires and provided a 12 h overnight urine sample. On average, women reported significantly more favourable mood states on standard psychometric tests, during the month of conception than during the previous non-conception cycles. In addition, they felt significantly less 'hassled' during the month of conception. However, mean urinary hormone excretion of adrenaline, noradrenaline and cortisol did not significantly differ between conception and non-conception cycles and there was little relationship between the psychological measures of mood state and excretion of adrenaline and cortisol. There was no evidence of increased coital frequency during the month of conception when mood states were improved, suggesting that stress effects on libido were unlikely to account for the findings. The results support the conclusion that psychosocial stress influences fertility in females but as yet mechanisms remain unclear. PMID- 9402305 TI - Semen donor recruitment strategies--a non-payment based approach. AB - Actual and projected prohibition of payment for semen donation in the UK and Canada has increased the need to examine alternative methods of donor recruitment. Evidence from a number of sources suggests that there is a large group of current and potential donors who are motivated more by meeting esteem needs than by payment. We develop an argument for using social marketing tools to create systematically an esteem-based approach to donor recruitment as an alternative to the payment approach. We conclude that esteem is a useful method of reciprocating the gift that donors make. PMID- 9402306 TI - Mitotic chromosomal anomalies among infertile men. PMID- 9402307 TI - DNA topoisomerase targeting drugs: mechanisms of action and perspectives. AB - The nuclear enzymes DNA topoisomerases I and II appeared as cellular targets for several antitumor drugs: campthotecin derivatives interacting with topoisomerase I, and actinomycin D, anthracycline derivatives, elliptinium acetate, mitoxantrone, epipodophyllotoxine derivatives, amsacrine and a new olivacine derivative, NSC-6596871 (S 16020-2), which interact with topoisomerase II. The functions of these enzymes are numerous and important since they are critical for DNA functions and cell survival. Despite the fact that they share the same target, topoisomerase II inhibitors have different mechanisms of action. Two principle types of induced alterations are involved in cellular resistance to topoisomerase II drugs: qualitative or quantitative alteration of the enzyme and/or increased drug efflux due to overexpression of P-glycoprotein. S 16020-2, a new olivacine derivative with a high antitumor activity against solid tumors, shows a potent cytotoxic effect against tumor cells expressing P-glycoprotein. This observation suggests that the comprehension of the respective effects of topoisomerase inhibitors and the precise knowledge of their mechanisms of resistance would improve the use of this therapeutic class in the clinic within rational chemotherapeutic combinations. PMID- 9402308 TI - A phase I study of ambulatory continuous infusion paclitaxel. AB - Chemotherapy given by continuous infusion may have different toxicity profiles and efficacy than when given by bolus administration. Thirty-one patients with refractory tumors entered a phase I trial in which paclitaxel was administered for 7 days by continuous infusion every 28 days. Only one patient required hospitalization for treatment, because of an initial poor performance status, and most carried out normal activities on an ambulatory basis. After the first three patients, patients were entered in cohorts of five with the starting dose of 120 mg/m2. Each subsequent cohort was begun at a dose 10% higher than the previous cohort. Later courses within each cohort were increased 10% in an individual patient, if toxicity allowed. Nausea was rare. Of 15 patients with a soft tissue sarcoma refractory to doxorubicin, dacarbazine, ifosfamide, and etoposide, there were: one partial response (PR), five stable diseases and eight progressive; one patient was not evaluable for response. The PR occurred in a patient with a very aggressive sarcoma and very bulky disease, and was maintained for more than 1 year. We conclude that paclitaxel given by ambulatory continuous i.v. infusion is well tolerated with a maximally tolerated starting dose of 160 mg/m2. PMID- 9402309 TI - Phase I study of VRCTC-310, a purified phospholipase A2 purified from snake venom, in patients with refractory cancer: safety and pharmacokinetic data. AB - A phase I study was performed to evaluate the maximum tolerated dose (MTD), safety profile and pharmacokinetic data with VRCTC-310, a natural product derived from purified snake venom fractions, with phospholipase A2 activity and inhibitory effects against human and murine tumor cell lines. Fifteen patients with refractory malignancies were entered after providing written informed consent. VRCTC-310 was administered as an intramuscular injection daily for 30 consecutive days. Doses were escalated from 0.0025 to 0.023 mg/kg. Toxicities included local pain at the injection site, eosinophilia, reversible diplopia and palpebral ptosis. Dose escalation was stopped at 0.023 mg/kg, when two patients had developed anaphylactoid reactions. Both cases had high VRCTC-310-specific IgG by EIA. MTD was 0.017 mg/kg and the recommended dose for phase II studies is 0.017 mg/kg. Stabilization was found in six patients. PMID- 9402310 TI - Pharmacokinetics of methotrexate-albumin conjugates in tumor-bearing rats. AB - Linking chemotherapeutic drugs to a macromolecular carrier system may enhance tumor targeting, reduce toxicity and overcome drug resistance mechanisms. As an elementary model to evaluate the pharmacological properties of macromolecular drug carrier systems we chose rat serum albumin (RSA) for carrier and methotrexate (MTX) as antineoplastic drug. The conjugation procedure yielded conjugates with an approximate 1:1 molar loading rate (MTX(1)-RSA). In the first part of the study a residualizing [111In]DTPA protein label was used for mapping in vivo the catabolic sites of the native carrier protein and of the MTX(1)-RSA drug conjugate in Walker 256 carcinosarcoma bearing rats. The tumor accumulation was about 14% of the injected dose for the RSA and MTX(1)-RSA tracers after 24 h. Tracer entrapment by organs with an active mononuclear phagocyte system was low (liver below 7% and spleen below 1.5% of the injected dose after 24 h). The 1:1 conjugation of MTX to RSA did not decisively alter the pharmacokinetic properties nor the tumor or tissue distribution of the native carrier protein RSA. In the second part of the study the different properties of the MTX(1)-RSA conjugate were compared with MTX in vivo. About 2 mg MTX/kg body weight either of the drug conjugate or of the original drug were injected after being additionally spiked with radiolabeled tracers. Plasma concentrations were simultaneously determined by immunological and radioactive means. After 24 h about 12% MTX(1)-RSA was found in circulation compared to 0.03% MTX. Favorable tumor accumulation rates of about 14% were achieved for MTX(1)-RSA versus 0.04% for MTX. About 45-fold more of the injected dose of [3H]MTX accumulated in the liver as compared to the tumor (1.5 versus 0.03%) after 24 h. Conjugation of MTX to RSA reversed this ratio in favor of the tumor to 1:1.4 (13.6 versus 9.6%). In conclusion, the potential therapeutic benefit of the MTX(1)-RSA conjugate lies in its very long tumor exposure time and its improved tumor accumulation rate compared to conventional MTX. In addition the conjugation to albumin might enhance the therapeutic effects over those achieved by long-term continuous infusion of MTX, as MTX(1)-RSA enters the cells by a different uptake mechanism. This might also help to circumvent MTX resistance mechanisms, such as a reduction in folate receptor numbers or impaired MTX polyglutamylation. PMID- 9402311 TI - Cytotoxic and antitumor activity of MEN 10710, a novel alkylating derivative of distamycin. AB - MEN 10710 is a new synthetic distamycin derivative possessing four pyrrole rings and a bis-(2-chloroethyl)aminophenyl moiety linked to the oligopyrrole backbone by a flexible butanamido chain. Its biological properties have been investigated in comparison with the structurally related compound, tallimustine (FCE24517), and the classical alkylating agent, melphalan (L-PAM). Cytotoxic potency of MEN 10710 was increased from 10- to 100-fold, as compared to tallimustine or L-PAM in murine L1210, human LoVo and MCF7 tumor cell lines. MEN 10710 was still active against L1210/L-PAM leukemic cells, while a partial cross-resistance was observed in LoVo/DX and in MCF7/DX cells selected for resistance to doxorubicin and expressing a MDR phenotype. Treatment with verapamil (VRP) reduced the resistance to tallimustine, but not to MEN 10710, in MCF7/DX cells. The cytotoxic effects reflect in vivo antitumor potency and toxicity in the treatment of human tumor xenografts. MEN 10710 was more effective in A2780/DDP, an ovarian carcinoma selected for resistance to cisplatin. On the other hand, the IC30 for inhibiting murine granulocyte/macrophage colony formation was 50 times higher for MEN 10710 than for tallimustine, suggesting a lower myelotoxic potential. In conclusion, the particular biological profile of MEN 10710 characterized by a marked cytotoxic potency, an interesting antitumor efficacy and a reduced in vitro myelosuppressive action may represent a further improvement in the rational design of a novel distamycin-related alkylating compound. PMID- 9402312 TI - Effects of cisplatin and taxol on inducible nitric oxide synthase, gastrin and somatostatin in gastrointestinal toxicity. AB - Cisplatin (9 mg/kg) or taxol (20 mg/kg) treatment of Wistar rats produced a sharp decrease in inducible nitric oxide synthase (iNOS) and gastrin in the pyloric region of the stomach, and an increase in iNOS and somatostatin in the pancreatic islets. Nitric oxide (NO) functions as a relaxation factor in the smooth muscle of the muscularis mucosa while gastrin plays an important role in the gastroprotection of the mucosa through NO. It is proposed that a decline of the iNOS and gastrin after cisplatin or taxol treatments is related to distention of the stomach, and possibly nausea and vomiting. Hyperglycemia and glucose intolerance after cisplatin treatment may be caused by increases of somatostatin and iNOS in the pancreatic islets. Combination therapy with cisplatin and taxol seems to ameliorate various toxicities due to these two individual drugs. PMID- 9402313 TI - In vitro toxicity studies with mitomycins and bleomycin on endothelial cells. AB - Pulmonary side effects are increasingly observed as dose-limiting toxicity (DLT) of cancer treatment. The available preclinical models have a limited predictive value for lung toxicity in humans. We have attempted to elucidate potential mechanisms involved in these reactions, by studying the effects on cells, possibly involved in these reactions after in vitro exposure to drugs with known lung toxic effects. We have investigated the effects of bleomycin (BLM), mitomycin C (MMC), KW-2149 and its two known metabolites, M16 and M18, on oxygen radical production by granulocytes, on cytokine production: interleukin (IL)-6, transforming growth factor (TGF)-beta, tumor necrosis factor (TNF)-alpha by a human macrophage cell line (THP-1), by human endothelial cells (HVEC and HMEC) and a human colorectal cancer cell line (DLD-1), and on the cytotoxicity on endothelial cells in both confluent and non-confluent culture. The generation of oxygen radicals by normal and pre-stimulated granulocytes was not increased after preincubation with any of the drugs, at the concentrations tested. None of the cytokines (IL-6, TNF-alpha or TGF-beta) was found significantly increased in culture medium after exposure to any of the mitomycins. This was in contrast with the effect of BLM incubation, causing a rise in TGF-beta concentration. Both types of endothelial cells showed a dose-dependent, exposure duration-dependent, proliferation inhibition for all agents tested. This inhibitory effect was clearly proliferation dependent as shown by the increased inhibition in semi confluent as opposed to confluent endothelial cell cultures. Both mitomycins tested were more cytotoxic than BLM to both confluent and proliferating endothelial cells. PMID- 9402314 TI - Comparative effect of verapamil, cyclosporin A and SDZ PSC 833 on rhodamine 123 transport and cell cycle in vinblastine-resistant Chinese hamster ovary cells overexpressing P-glycoprotein. AB - The product of the mdr1 gene, P-glycoprotein (P-gp), represents a common mechanism of cellular resistance to a wide variety of structurally and functionally unrelated drugs. A range of structurally different P-gp inhibitors, such as verapamil, cyclosporin A and SDZ PSC 833, have been shown to modify multidrug resistance (MDR). We used flow cytometry to investigate in vitro modulation of P-gp-dependent efflux of rhodamine 123 (Rh123). The capacity to modulate the MDR phenotype of vinblastine-resistant Chinese hamster ovary (CHO) cells was assessed by analyzing the concentration of modulator needed to decrease the Rh123 mean fluorescence intensity by 50%. We found that the cyclosporin derivative SDZ PSC 833 was significantly more effective than cyclosporin A and verapamil, either in the presence or absence of fetal calf serum-supplemented media. This study indicates that analysis of Rh123 efflux modulation can be used to determine the optimal doses of MDR inhibitors in vitro and suggests that more than one modulator is needed to measure P-gp function, since verapamil had no effect on Rh123 modulation when MDR cells were used. PMID- 9402315 TI - Antitumor activity of oxaliplatin in combination with 5-fluorouracil and the thymidylate synthase inhibitor AG337 in human colon, breast and ovarian cancers. AB - Oxaliplatin, classical [5-fluorouracil (5-FU)] and non-classical (AG337) thymidylate synthase inhibitors have shown promising activity in the treatment of cancer. This study investigates the cytotoxic effects of oxaliplatin in combination with 5-FU and AG337 in cultured human colon (HT29, CaCo2), breast (MCF-7, MDA-MB-231) and ovarian (2008) cancer cell lines, and their derived counterparts selected for their resistance to 5-FU (HT29-5-FU), doxorubicin (MCF 7mdr) or cisplatin (2008C13). Therapeutic experiments were conducted in mice bearing colon-HT29 xenografts and in the GR hormone-independent mammary carcinoma model. In vitro, oxaliplatin shows potent cytotoxic activity in colon (IC50 from 2.1 +/- 1.1 to 5.9 +/- 1.7 microM), ovarian (IC50 = 10 +/- 1.6 microM) and breast cancer cells (IC50 from 7.4 +/- 2.7 to 17.9 +/- 7.1 microM). Oxaliplatin was a potent inhibitor of DNA synthesis and bound to cellular DNA. Surprisingly, the overall amount of oxaliplatin DNA binding was significantly inferior to that induced by isocytotoxic concentrations of cisplatin in HT29 (p=0.026). In vitro, synergistic antiproliferative effects were observed when oxaliplatin was added to 5-FU and AG337. Those synergistic effects of combinations were maintained in colon HT29-5-FU cancer cells. In vivo, 5-FU increased significantly the antitumor activity of oxaliplatin in HT29 xenografts (p=0.0036), and similarly 5-FU and AG337 increased the activity of oxaliplatin in the GR tumor model (p=0.0012). These data may encourage further clinical investigation of oxaliplatin in combination with classical and non-classical thymidylate synthase inhibitors in the treatment of human cancers. PMID- 9402316 TI - The mechanism of locally enhanced production of tumor necrosis factor-alpha in tumor tissues by the administration of a new synthetic lipid A analog, ONO-4007, in hepatoma-bearing rats. AB - ONO-4007 is a new synthetic lipid A analog with low endotoxic activities. We previously found that ONO-4007 induced the production of tumor necrosis factor (TNF)-alpha in rat hepatoma KDH-8 tumor tissues and brought about the regression of transplanted KDH-8 cells. By contrast, ONO-4007 did not induce TNF-alpha production in spleens and sera 90 min after treatment. In the present study we attempted to elucidate how ONO-4007 induces TNF-alpha production in tumor tissues locally. We found that extracellular matrix including gelatin, fibronectin and Matrigel did not induce TNF-alpha production in splenocytes treated with ONO-4007 in vitro. However, splenocytes co-cultured with cKDH-8/11 tumor cells in the presence of ONO-4007 produced more TNF-alpha than splenocytes cultured by themselves in the presence of ONO-4007. The stimulation of cKDH-8/11 cells in the presence of ONO-4007 for splenocytes to produce TNF-alpha depended on the type of contact between the cells. The cKDH-8/11 cells fixed in formalin were not able to induce TNF-alpha production of splenocytes even in the presence of ONO-4007. However, syngeneic fibrosarcoma cell line KMT-17/A3, allogeneic hepatocellular carcinoma cell line LDH and rat lung endotherial cell line RLE induced TNF-alpha production in splenocytes, but their stimulation was weaker than that of cKDH 8/11. The soluble form of the cKDH-8/11 cell membrane did not stimulate splenocytes to produce TNF-alpha in the presence of ONO-4007. cKDH-8/11 cells did not stimulate the splenocytes devoid of macrophages to produce TNF-alpha in the presence of ONO-4007. PMID- 9402317 TI - Prevention of peritoneal metastasis of cancer with dextran sulfate--an experimental study in mice. AB - Peritoneal metastases occur most often in the greater omentum, where tumor implantation sites are densely distributed. We used dextran sulfate (S-Dex) as an anti-cell-adherence agent to prevent i.p. seeded malignant cells from causing peritoneal metastases. S-Dex was tested for its anti-adherent activity against B 16 melanoma cells on plastic, and was examined for its ability to prevent implantation in the omentum and to improve survival in mice after B-16 melanoma was inoculated i.p. S-Dex prevented B-16 melanoma cells from adhering to the plastic wall. S-Dex reduced the number of B-16 melanoma cells implanted into the greater omentum and improved the survival of mice inoculated with B-16 melanoma cells. We conclude that S-Dex attenuated peritoneal metastases when B-16 melanoma cells were seeded i.p. PMID- 9402318 TI - Therapeutic effects of a new synthetic lipid A analog, ONO-4007, on rat hepatoma KDH-8 depend on tumor necrosis factor-sensitivity of the tumor cells. AB - ONO-4007 is a new synthetic lipid A derivative with low endotoxic activities. ONO 4007 was effective against KDH-8, a tumor necrosis factor (TNF)-sensitive rat hepatoma cell line, but neither effective against KMT-17, a TNF-resistant rat fibrosarcoma cell line, nor SST-2, a TNF-resistant rat mammary adenocarcinoma cell line. We have established two sublines from KDH-8 to further examine the therapeutic mechanisms of ONO-4007 in vivo: TNF-sensitive KDH-8/YK and TNF resistant cKDH-8/11. The two sublines equally proliferated in vitro. Multiple systemic i.v. administration of ONO-4007 was performed on days 7, 14 and 21 after tumor implantation. Although treatment with ONO-4007 had no effect on the growth of cKDH-8/11 in WKAH rats in vivo, 60% of KDH-8/YK-bearing rats treated with ONO 4007 survived. The administration of ONO-4007 brought about significant therapeutic effects on KDH-8/YK-bearing rats but not on cKDH-8/11-bearing rats. These results suggest that ONO-4007 is therapeutically useful for the treatment of TNF-alpha-sensitive tumors. PMID- 9402319 TI - Reed-Sternberg cells and the TNF family of receptors/ligands. AB - Treatment of cancer with means other than chemo- and radiation therapy becomes more and more important. Through the better understanding of tumor biology approaches towards the cure of cancer interfering with the pathophysiological mechanisms of malignancy can be considered. Hodgkin's disease is a good example for the role of the immune system in cancer. The Reed-Sternberg (RS) cells, malignant cells of Hodgkin's disease (HD), are surrounded by tumor infiltrating lymphocytes (TILs) and still evade immunesurveillance. In this respect the importance of the superfamily of tumor necrosis factor (TNF) receptors and ligands is becoming more and more clear. Ligand-receptor interaction either leads to death or survival signals. Many of these receptors and ligands are expressed by the RS cells and the surrounding lymphocytes. Their expression and function in HD are discussed and future directions for possible therapeutical investigations are proposed. PMID- 9402320 TI - The Wilms tumour gene WT1 in leukaemia. AB - The WT1 gene is essential for kidney development and is mutated in some Wilms tumours. It is also expressed at a high level in many acute leukaemias and in some haematopoietic progenitor cells, and mutations have been found in leukaemias. The function of WT1, which is a zinc finger protein and has domains characteristic of transcription factors, is not well understood. The level of expression is highest in leukaemias with immature phenotypes. Expression of WT1 is downregulated during differentiation of leukaemic cell lines and high levels of WT1 expression can cause cell cycle arrest and/or apoptosis. This may reflect a role in the control of normal haematopoiesis, which can be abrogated by mutations in the gene and form part of the pathway towards leukaemogenesis. PMID- 9402321 TI - Fibroblast growth factors and early hemopoietic cell development. AB - The role of fibroblast growth factors (FGFs) in the regulation of the human hematopoiesis is controversial. Older publications suggest that FGFs play an important role in regulating the proliferation of human hematopoietic progenitor and stem cells. Such studies should be interpreted with caution because they were typically carried out in the presence of serum which not only contains significant amounts of FGFs, but other cytokines as well. The development of greatly improved techniques for isolating purified populations of primitive hematopoietic cells, for culturing such cells in serum free conditions, and for analyzing the molecular consequences of exposing these cells to FGF cytokines allow the conclusions of these earlier studies to be tested more rigorously. Our investigations, and those of others, suggest that FGFs may not play an important direct role in regulating the development of human hematopoietic stem cells. These data, and our view of their implications, are discussed in this review. PMID- 9402322 TI - Clonal populations of T-cells in patients with B-cell malignancies. AB - Patients with B-cell malignancies are often immunosuppressed and have defective T cell function in vitro. In addition they frequently have unusual T-cell populations in the peripheral blood including an increase in the number of activated T-cells and an inverted CD4:CD8 ratio. More recently several reports have documented the presence of large, monoclonal populations of T-cells in patients with paraproteinaemia, B-CLL and hairy cell leukemia. Such cells can reach very high levels in the peripheral blood, occasionally representing over 50% of all CD8+ T-cells. These clonally expanded cells have a characteristic morphology and phenotype in that they are often large, granular cells with natural killer cells markers. Their properties have not been studied in detail but they appear to suppress immunoglobulin production and kill cell targets in an MHC-unrestricted manner. The relationship of clonal T-cells to the B-cell tumour is unclear. They may be directly interacting with the malignant clone or alternatively be nonspecifically activated secondary to a disruption of the immune homeostasis by tumour cells. If they indeed represent an attempt by the immune response to control the malignant cells it is possible that they may be utilised in future attempts at immunotherapy. PMID- 9402323 TI - The granulocyte colony-stimulating factor (G-CSF)/G-CSF receptor (G-CSFR) system in B-cell chronic lymphocytic leukemia. PMID- 9402324 TI - Characterization of the novel focal adhesion kinase RAFTK in hematopoietic cells. AB - Protein tyrosine kinases (PTKs) mediate signals that respond to many pivotal cellular functions. Tyrosine phosphorylation, controlled by the coordinated actions of protein tyrosine phosphatases (PTPs) and PTKs, is a critical control mechanism for various physiological processes, including cell growth, differentiation, metabolism, cell cycle regulation and cytoskeleton function. The focal adhesion kinase (FAK) is a widely expressed non-receptor tyrosine kinase that is implicated in integrin-mediated signaling and plays a role in signal transduction pathways mediating cell adhesion, motility and anchorage-independent growth. Recently, we and others have identified a novel protein tyrosine kinase termed RAFTK, (also known as Pyk2 or Cak-beta), which is related to FAK. This review describes the role of RAFTK in various signaling cascades mainly in reference to hematopoietic cell lineages. PMID- 9402325 TI - P-glycoprotein expression in de novo acute myeloid leukemia. AB - Detection of the multidrug resistance P-glycoprotein (PGP) phenotype was performed at the time of diagnosis in 223 patients with acute myeloid leukemia (AML) by flow cytometry using C219 Monoclonal Antibody (MoAb). On the other hand, JSB1 MoAb was tested in 173 of these samples. At onset, PGP was detected in 57.4% of cases with C219 and 75.9% of cases with JSB1. There was no correlation between PGP expression and sex, age, marrow blast percentage or extramedullary disease. On the contrary, strict correlations were noted either between C219 negativity and FAB M3 subtype or between C219 positivity and FAB M5 group (P = 0.003). Significant correlation was found between PGP phenotype and CD7, as 143 of 223 samples had similar patterns of staining with C219 (P < 0.0001). Finally, there was a close relationship between C219 and JSB1 positivity: all the C219+ cases were positive for JSB1 (P < 0.0001). Concerning the karyotype, most patients with monosomy or del (7) were MDR positive; on the other hand, most patients with t(8;21) or t(15;17) were MDR negative. Rh123 accumulation studies showed a significant decrease of mean fluorescence intensities both in C219 and in JSB1 positive cases in comparison with PGP negative ones (P < 0.001). A significant decrease of remission induction rates (CR) was highlighted both between C219+ and C219- and between JSB1+ and JSB1- cases (32.1% v 62.1% and 32.6% v 73.8%, respectively, with P < 0.0001). The overall survival and the remission duration (CCR) were significantly shorter both in C219+ and in JSB1+ patients with no relationship to age. Furthermore, a higher rate of early relapses was noted among MDR+ when compared with MDR- patients both for C219+ and JSB1+ cases. The combination (C219- JSB1+) identified a subset of patients with an intermediate prognosis. On multivariate analysis, C219 and JSB1 were confirmed to be independent prognostic factors for achievement of CR, overall survival and CCR. In conclusion, the assessment of MDR phenotype by flow cytometry is a crucial prognostic factor of treatment outcome in AML. PMID- 9402326 TI - Large scale manufacturing of TXU(anti-CD7)-pokeweed antiviral protein (PAP) immunoconjugate for clinical trials. AB - We have conjugated the murine monoclonal anti-CD7 antibody TXU to the plant hemitoxin pokeweed antiviral protein (PAP) to construct an effective immunotoxin against CD7 antigen positive hematologic malignancies. The scaled-up production and purification of TXU antibody, PAP toxin, and TXU-PAP immunotoxin permitted the manufacturing of a highly purified clinical-grade TXU-PAP preparation. In clonogenic assays, TXU-PAP elicited selective and potent cytotoxicity against CD7 antigen positive human leukemia cells and killed primary clonogenic leukemic cells from T-lineage acute lymphoblastic leukemia (ALL) patients. To our knowledge, this pre-IND work represents the first effort of producing a clinical grade PAP immunotoxin for treatment of T-lineage ALL. Since the CD7 antigen is also expressed on AML cells, TXU-PAP could also be useful for the treatment of CD7 positive acute myeloid leukemia (AML) patients. PMID- 9402327 TI - Benign and malignant smooth muscle tumors containing Epstein-Barr virus in children with AIDS. AB - Smooth muscle tumors (leiomyosarcomas) are the second most prevalent malignancy of children with the acquired immunodeficiency syndrome (AIDS). We have investigated the tumors, plasma, and peripheral white blood cells of eight children with AIDS with smooth muscle tumors for evidence of tumor association with human immunodeficiency virus (HIV) and Epstein-Barr virus (EBV). Very low levels of HIV were found in the tumors of the AIDS patients, probably resulting from blood-borne carriage of virus. These smooth muscle tumors had very high quantities of EBV in all the tumor cells by in situ hybridization, with an average of 4.5 EBV genomes per cell by quantitative polymerase chain reaction amplification. Increased amounts of EBV were found in the peripheral blood cells of two AIDS patients before the time of tumor diagnosis. EBV clonality studies demonstrated different monoclonal EBV infection of two separate colonic tumors from one patient, and dual or mixed monoclonal EBV infection in another patient. The muscle cells of leiomyomas and leiomyosarcomas of patients with AIDS demonstrated prominent staining with antibodies to the EBV receptor. The uniform distribution and striking amount of EBV in the tumor cells demonstrates that EBV is capable of infecting smooth muscle cells and that these cells support EBV replication. Clonal EBV proliferation suggests that EBV infection occurs at an early stage of tumor development. These findings indicate that EBV has a causal role in the oncogenesis of leiomyosarcomas of patients with AIDS. PMID- 9402328 TI - Incidence and impact of light chain associated (AL) amyloidosis on the prognosis of patients with multiple myeloma treated with autologous transplantation. AB - Little is known about the incidence of clinically occult AL amyloid in patients with multiple myeloma and its impact on prognosis of these patients. To address these issues, subcutaneous fat pad aspirates (SAFA) and bone marrow biopsies were evaluated for the presence of amyloid in a cohort of newly diagnosed patients with multiple myeloma prior to enrollment on a phase II study including tandem transplants. Organ directed biopsies were performed when clinically indicated. Presence of amyloid at > or = 1 site was noted in 32 patients (38%). SAFA was positive in 25 (31%), bone marrow in 8 patients (10%) and other organ sites in 7 patients. Patients with and without amyloid did not differ in disease characteristics, in particular no lambda predominance was observed in patients with amyloid. Event free survival (59+ vs 52 months; p = .9) and overall survival (59+ vs 66+ months; p = .9) were similar in both groups. Even the seven patients with symptomatic organ involvement with AL amyloid had a median overall survival of 38+ months. In conclusion, AL amyloidosis occurs more often than previously reported, but its presence does not influence the outcome of these patients after transplantation. PMID- 9402329 TI - Pharmacokinetics of high-dose cytarabine and its deamination product--a reappraisal. AB - Cytarabine is intracellularly activated and correlations have been established between the pharmacokinetic behaviour of active metabolites and their antileukemic effect. Recently, a good response to high-dose treatment of leukemias has additionally been attributed to a so-called low deamination phenotype of cytarabine inactivation. Consequently, these findings would support plasma level monitoring of cytarabine and its metabolite uracil arabinoside in high-dose cytarabine regimens. This pharmacokinetic study presents data attempting to reevaluate these observations. Thirty-seven patients were treated by 3-h high-dose cytarabine infusions (9 patients 1000 mg/m2, 28 patients 3000 mg/m2) as part of their treatment for acute leukemia. Serial blood samples during and post infusion were analysed for cytarabine (araC) and its deamination product uracil arabinoside (araU) using HPLC with UV-detection. Considerable interindividual variation was observed in end-infusion plasma concentrations of araC (1000 mg/m2: 2.1-fold, 3000 mg/m2: 5.5-fold) and araU (1000 mg/m2: 2.7-fold, 3000 mg/m2: 2.9-fold). The median ratio of end infusion concentrations araU/araC (on a molar basis) was 5.6 (S.D. 3.0), extreme ratio values were 2 and 14. No differences of the araU/araC ratio were found between the two dosages used. Minimum plasma araC concentrations at the end of infusion were 10.5 micromol/l and 22.0 micromol/l at a dose of 1000 and 3000 mg/m2, respectively. In our European study population a "fast" deamination phenotype of cytarabine (araU/araC ratio > 14) was not be observed. PMID- 9402330 TI - Rearrangements of the BCL6 gene and chromosome aberrations affecting 3q27 in 54 patients with non-Hodgkin's lymphoma. AB - Chromosome aberrations affecting 3q27 are among the most frequent non-random abnormalities in non-Hodgkin's lymphomas (NHL), especially the diffuse, large cell type. Recently, an association between BCL6 rearrangement and frequent extranodal lesions, rare bone marrow infiltration and a favorable clinical outcome was reported. We performed molecular studies of the BCL6 gene in 54 patients with NHL. Twelve patients (22%) with rearranged BCL6 genes were selected for histological, clinical, molecular, and cytogenetic studies. Ten of these cases were diffuse, large cell type lymphoma, one a follicular lymphoma, and one a mantle cell lymphoma (MCL). All cases were of the B-cell type and this is the first time a rearranged BCL6 gene has been found in an MCL. Cytogenetic data for 10 cases were available and the partner sites of the 3q27 translocation were determined in 7 of 10 patients. These locations were variable, including 6p21.3, 9p22, and 14q11 in addition to the immunoglobulin loci 14q32 (IGH), 2p12 (IGK), and 22q11 (IGL). The heterogeneity in partner sites is distinct from other lymphoma subgroups and may suggest that the genetic events are not uniform among patients with BCL6 rearrangements. PMID- 9402331 TI - Eosinophilia associated with clonal T-cell proliferation. AB - Eosinophilia associated with the expansion of cloned T-cells is reviewed in relation to cytokine production. It has been proved that eosinophilopoiesis is caused by eosinophil-stimulating cytokines, including interleukin-5 (IL-5), granulocyte-macrophage colony-stimulating factor and interleukin-3, which are secreted from T-cells. Recently, we and other groups have reported several cases of eosinophilia including hypereosinophilic syndrome (HES) accompanied with proliferation of abnormal T-cells with an unusual phenotype CD3- CD4+ or CD3+ CD4 CD8- in the peripheral blood. The T-cells clonally proliferate, as confirmed by clonal rearrangements of the T-cell receptor (TCR) gene, and produce eosinophil stimulating cytokines, especially IL-5, with or without stimulation in vitro. Although HES is defined by the combination of unexplained prolonged eosinophilia and evidence of organ involvement, these observations suggest that increased production of eosinophil-stimulating cytokines from the abnormal T-cells with phenotype CD3- CD4+ or CD3+ CD4- CD8- may cause eosinophilia, some of which have been diagnosed as HES. PMID- 9402332 TI - Viability and quantification of progenitor cells in venesected blood from patients receiving escalated-dose epirubicin and cyclophosphamide with G-CSF for lymphoma: potential role in further increasing dose-intensity. AB - We assessed the concentration of haemopoietic progenitors in peripheral blood in six patients with de novo intermediate grade non-Hodgkin's lymphoma receiving multiple cycles of escalated dose epirubicin and cyclophosphamide on day 1 followed by 5 microg/kg of G-CSF (filgrastim) on days 2-14. Specimens were taken at days 12, 15 and 18 in cycles 1 and 2 and on day 15 for cycles 3-6. Progenitor numbers were maximal on day 15 in cycles 1 and 2. The median number of granulocyte-macrophage colony forming cells (GM-CFC) and CD34+ cells on day 15 of cycles 1 and 2 was 3.8 x 10(4)/ml and 11 x 10(4)/ml, respectively. A 600 ml venesection at this time would contain a median of 36 x 10(4) GM-CFC/kg (range 25 47) and 1.04 x 10(6) CD34+ cells/kg (range 0.73-1.4), based on individual patient weights. Day 15 progenitor numbers were maintained for the first 3 cycles but tended to fall thereafter. The viability of the progenitors collected in whole blood and stored at 4 degrees C for various time intervals was also assessed. The median percent of GM-CFC and erythroid blast forming units (BFU-e) surviving after storage for 48 hrs was 79% and 69% respectively and after 72 hrs was 48% and 63% respectively. Serum collected 2 hrs after the completion of chemotherapy had minimal inhibitory effect on progenitors collected prior to treatment. Our data demonstrate that two weeks after anthracycline-based chemotherapy and G-CSF in previously untreated patients the peripheral blood contains large numbers of progenitors. A 600 ml venesection at this time stored at 4 degrees C, and then reinfused after the next cycle of chemotherapy would contain sufficient viable progenitors to potentially hasten haematological recovery. PMID- 9402333 TI - Treatment of AL-amyloidosis with dexamethasone plus alpha interferon. AB - Current therapy for primary systemic (AL) amyloidosis has only modest efficacy (response rate 25%) and because it includes alkylating agents, it has a significant leukemogenic potential (actuarial risk 21% at 3.5 years). We treated 9 consecutive patients with biopsy proven AL amyloidosis seen at our institution with pulse dexamethasone induction (40 mg on days 1-4, 9-12, 17-20 repeated q 35 days) for 3-6 cycles followed by maintenance alpha interferon 3-6 million units thrice weekly. Three patients also received maintenance dexamethasone (40 mg/day x 4 days q 4-8 weeks) for the first year. Improvement in > or = 1 AL organ involvement was seen in 8 of 9 patients. Of 7 patients with nephrotic range proteinuria, 6 had > or = 50% reduction in nonspecific proteinuria with a median time to response of 4 months (range 3-9 months). Marked improvement in organ function was also seen in 4 patients with gastrointestinal, hepatic and neuropathic involvement. However, none of the 2 patients with congestive heart failure improved. This dexamethasone plus alpha interferon regimen, devoid of leukemogenic potential, may lead to rapid and durable improvement in organ function in a significant proportion of patients with AL amyloidosis and deserves further evaluation as front line therapy. PMID- 9402334 TI - A translocation t(8;14) and c-myc gene rearrangement associated with the histological transformation of B-cell acute lymphocytic leukemia (FAB-L2) into Burkitt's type (FAB-L3) leukemia. AB - We report a case of B-cell acute lymphocytic leukemia which showed histological transformation from an FAB-L2 into a Burkitt's type (FAB-L3). Both leukemias had identical immunoglobulin heavy-chain joining gene and kappa light-chain joining gene rearrangements, indicating the clonal identity of the two leukemias. A chromosomal analysis of leukemia cells on the onset indicated normal karyotype, whereas that of the transformed FAB-L3 showed t(8;14)(q24;q32). Furthermore, the proto-oncogene c-myc was in the germline configuration in the initial leukemia but in the rearranged configuration after transformation. Presence of t(8;14)(q24;q32) and the c-myc gene rearrangement after transformation suggested that the chromosomal translocation followed by the activation of the c-myc proto oncogene might be involved in the Burkitt's type transformation of the FAB-L2 leukemic clone, but not in the leukemogenesis of the initial FAB-L2 leukemia. PMID- 9402335 TI - Chronic myeloid leukemia in a woman with papillary carcinoma of the thyroid treated with radioactive iodine. AB - A 48-year-old woman presented with chronic myeloid leukemia six years after beginning repeated radioiodine treatments for papillary carcinoma of the thyroid gland. The literature on the association of radioiodine and chronic myeloid leukemia is reviewed. PMID- 9402337 TI - Maintained all-trans retinoic acid therapy in a patient with pseudotumour cerebri despite aggravated symptoms. PMID- 9402338 TI - The role of platelets in antimicrobial host defense. PMID- 9402336 TI - Bilateral eyelid localisation of a lymphoplasmacytoid lymphoma. AB - Eyelid localisation of non-Hodgkin's lymphoma is rare, and even more so when it is bilateral. We report a 58 year-old man who presented with an eyelid localisation of lymphoplasmacytoid lymphoma. The initial treatment was chemotherapy with good improvement but the relapse lead us to give radiotherapy with no further relapse 20 months later. Radiotherapy is the current treatment of localised eyelid lymphomas with excellent results. The prognosis is related to the initial staging and the 10-year survival rate is close to 80%. PMID- 9402339 TI - Proceedings of the 2nd International Workshop on Helicobacter pylori Infections in the Developing World. Lima, Peru, 28-31 January 1996. PMID- 9402340 TI - Epidemiological features of Helicobacter pylori infection in developing countries. AB - Helicobacter pylori infection has a worldwide distribution, and it has distinct epidemiological features in developing countries. In contrast to that in developed countries, H. pylori infection in developing countries seems to be nearly universal, beginning in early childhood. Children become infected in the first few months of life; in some communities as many as 50% of the children are infected by the age of 5 years, and up to 90% are infected by the time they reach adulthood. In some developing countries with improvements in industrialization, socioeconomic conditions, and hygiene, infection rates are lower. The incidence of H. pylori infection, determined indirectly, also suggests a rate several times higher than that in developed countries. Marked differences in H. pylori seroprevalence have been observed between various ethnic and racial groups. Although the mode of transmission of H. pylori remains uncertain, evidence suggests person-to-person transmission occurs. PMID- 9402341 TI - Helicobacter pylori infection in Desert Storm troops. AB - To determine whether military personnel deployed outside the United States are at increased risk of Helicobacter pylori infection, we evaluated U.S. Army personnel who served in the Persian Gulf from August 1990 to April 1991. Of 204 subjects from whom paired predeployment and postdeployment serum specimens were obtained, 76 (37%) were seropositive for IgG antibody to H. pylori before deployment by an enzyme-linked immunosorbent assay. Of the 111 initially seronegative subjects evaluated before and after a 7.5-month deployment, five (4.5%) seroconverted. The calculated annual seroconversion rate was 7.3%. In a postdeployment questionnaire, 62% of soldiers reported an episode of diarrhea while deployed, but there was not an increased rate of diarrhea or upper gastrointestinal symptoms in soldiers who were infected before deployment or in those who seroconverted. These data suggest that the risk of H. pylori infection increases during long-term deployment and that acute infection is not distinguishable from other gastrointestinal illnesses encountered during deployment. PMID- 9402342 TI - Helicobacter pylori infection in Chile. AB - This article summarizes studies designed to evaluate the role of Helicobacter pylori infection in Chile, described in 21 reports from nine centers in various Chilean regions published between 1985 and 1995. According to their data, H. pylori infection is quite frequent among patients with a variety of gastric conditions, including adults (43%-92%) and children (6%-100%). Levels of specific IgG antibodies to H. pylori are also elevated among patients with duodenal ulcers (100%) and gastritis (86%) as well as asymptomatic adults (75%). Combination therapy with three (but not two) drugs has been proved effective, with clinical improvement, ulcer cure, and H. pylori eradication occurring in well-controlled studies. Available evidence suggests that antibiotic resistance is not a major problem in treatment. The H. pylori reinfection rate is low (4.2% per year), suggesting that combination therapy with three drugs constitutes a cost-effective alternative for treating colonized symptomatic patients. Concurrent preliminary studies revealed that antibodies to VacA but not CagA proteins correlate with disease severity in Chilean patients. It can be concluded that local research assists local administrators of health resources to implement adequate policies to prevent, control, and treat H. pylori-related pathologies. PMID- 9402343 TI - Helicobacter pylori: prevalence, transmission, and serum pepsinogen II concentrations in children of a poor periurban community in Bangladesh. AB - The aim of this study was to determine the age-specific prevalence of Helicobacter pylori infection in infants and children aged 1-99 months from a poor periurban community in Bangladesh. We also examined the frequency of infection among infants and their 53 immediate family members and evaluated the relationship between infection and fasting serum group II pepsinogen (pepsinogen II) concentration in 76 children. Sixty-one percent of 1-3 month-old infants tested positive for H. pylori; this rate declined steadily to 33% in children aged 10-15 months and then increased to 84% in children aged 5-8 years. The H. pylori infection rate was 2.5 times higher in children with illiterate mothers. No difference in infection rate was detected among the family contacts of infected vs. noninfected infants. H. pylori-infected children had significantly higher serum pepsinogen II concentrations than did noninfected children (P < .001). We conclude that infection with H. pylori is highly prevalent and occurs at an early age. An environmental factor or factors, rather than or in addition to intrafamilial spread of this infection, are important in poor communities of Bangladesh. The higher levels of serum pepsinogen II in H. pylori-positive children might indicate the presence of gastritis in such asymptomatic children. PMID- 9402345 TI - Nondispersive infrared spectrometry: a new method for the detection of Helicobacter pylori infection with the 13C-urea breath test. AB - Nondispersive infrared spectrometry (NDIRS) was used to detect Helicobacter pylori infection with the 13C-urea breath test. The results were compared with those of standard isotope ratio mass spectrometry (IRMS). Both methods accurately distinguished between H. pylori-positive and H. pylori-negative individuals. The results demonstrate that NDIRS technology is accurate and therefore of equal value to standard IRMS for detection of H. pylori infection. It can be recommended for routine clinical application. As NDIRS technology is much cheaper than current IRMS machines, we consider the new method extremely useful for clinical applications. PMID- 9402344 TI - Helicobacter pylori populations in Peruvian patients. AB - Helicobacter pylori is an extremely diverse species. The characterization of strains isolated from individual patients should give insights into colonization and disease mechanisms and bacterial evolution. We studied H. pylori isolates from patients in the Japanese-Peruvian Polyclinic in Lima, Peru, by determining metronidazole susceptibility or resistance and by random amplified polymorphic DNA (RAPD) fingerprinting (a measure of overall genotype). Strains isolated from several biopsy specimens from each of 24 patients were studied. Both metronidazole-susceptible and -resistant strains were isolated from 13 patients, whereas strains of more than one RAPD type were isolated from only seven patients. We propose that the homogeneity in RAPD fingerprints for strains isolated from most persons reflects selection for particular H. pylori genotypes during chronic infection in individual hosts and the human diversity in traits that are important to this pathogen. Carriage of related metronidazole-resistant and -susceptible strains could reflect frequent metronidazole use in Peru and alternating selection for resistant and susceptible phenotypes during and after metronidazole therapy. PMID- 9402346 TI - Chronic atrophic gastritis: early diagnosis in a population where Helicobacter pylori infection is frequent. AB - Chronic atrophic gastritis (CAG) is a premalignant condition characterized by loss of gastric antral deep glands. The histologic changes in antral gastric biopsy specimens from 54 Peruvian patients with dyspepsia were studied to detail the development and characteristics of CAG. Ninety-six percent of the biopsies revealed severe superficial mucosal inflammation and 89% showed deep inflammation. Moderate or severe CAG was present in 36 (67%) of the 54 patients. In the early stages of CAG, a glandular lymphoid adherence lesion was noted in 17 (31%) of the 54 biopsy specimens. This lesion consisted of lymphocytes adherent to the antral deep gland cells and was associated with glandular epithelium alterations. The late stage was characterized by small glands, remnants of glands, and gland replacement with a fibrocellular infiltrate or intestinal metaplasia. We propose that the development of CAG probably proceeds via a stereotyped sequence, with an early deep inflammatory component that may trigger local gland destruction and eventual permanent loss. PMID- 9402347 TI - Geographic factors probably modulating alternative pathways in Helicobacter pylori-associated gastroduodenal pathology: a hypothesis. AB - It is hypothesized that probable geographic factors of nutritional type, nonrelated to development or socioeconomic level, may modulate the conversion of Helicobacter pylori-associated active chronic gastritis from its early stages to chronic atrophic gastritis (CAG). The factors could be diets low in antioxidant vitamins and other micronutrients such as selenium. In regions of the world where these modulating factors are not present, active chronic gastritis tends to stay in its early stages and to predispose individuals to duodenal ulcer. On the contrary, in regions where the modulating factors are present, the frequency of CAG increases markedly. When CAG becomes severe and extensive, hypochlorhydria ensues. Hypochlorhydria decreases the predisposition to duodenal ulcer, while CAG, a precancerous lesion, predisposes individuals to gastric cancer of the intestinal type. The hypothesis could be tested in a multicenter, multiregional study to (1) determine endoscopically and histologically the prevalence rates of duodenal ulcer, gastric ulcer, gastric cancer, and H. pylori-associated CAG in large series of dyspeptic patients and (2) correlate these prevalence rates with blood levels of micronutrients in these patients. PMID- 9402348 TI - Helicobacter pylori gastritis, peptic ulcer, and gastric cancer: clinical and molecular aspects. AB - Helicobacter pylori causes specific ultrastructural changes to the gastric mucosa. In developing countries a high percentage of infants acquire this infection, which initially causes a transient drop in stomach acid and thus allows transit of lower bowel pathogens, with consequent diarrhea and malnutrition. When infection occurs at an early age, the acid-producing cells of the stomach are involved in the inflammation, and the lifelong reduced acid output means a duodenal ulcer rarely develops. However, lifelong gastric inflammation leads in due course to atrophy, and in the presence of other factors gastric cancer may develop. People infected with H. pylori on average are of shorter stature than uninfected people. Adherence of H. pylori to the gastric mucosa is a prerequisite for infection, and a new binary model of adherence has been shown recently. Chaperonins of H. pylori induce macrophages to secrete cytokines, which leads to an immunologic cascade and inflammation. PMID- 9402349 TI - Role of Helicobacter pylori infection in the development of pernicious anemia. AB - It is now accepted that most patients with atrophic gastritis of the stomach have been infected with Helicobacter pylori. Several investigators have also suggested the possibility that H. pylori is involved in the early stages of pernicious anemia, which leads to severe atrophic gastritis of the fundus. In this article, studies investigating the association of this specific form of atrophic gastritis and H. pylori infection are reviewed. Most of the published studies indicate that patients with pernicious anemia are infected with H. pylori less often than are age-matched controls. However, because H. pylori infection may be present before the development of pernicious anemia, prospective studies during the pre pernicious anemia stage of gastritis are needed. PMID- 9402350 TI - Antimicrobial treatment of Helicobacter pylori infection. AB - Helicobacter pylori is susceptible to many antimicrobials, but clinically only a few are effective. Two antimicrobials with bismuth or ranitidine or a proton pump inhibitor such as omeprazole are required to achieve a cure rate of >90% and to avoid resistance, which occurs when clarithromycin or metronidazole is the single antimicrobial used. Bismuth plus metronidazole and tetracycline is effective but causes more side effects than does treatment with omeprazole, amoxicillin, and clarithromycin; metronidazole can replace clarithromycin. To ensure a high cure rate, treatment is required for 10 days, but 7-day regimens have sometimes been as successful. A course of ranitidine bismuth citrate for 28 days, given with clarithromycin for the first 14 days, cures 80%-85% of patients, but given with amoxicillin it cures only 74%. In developing countries resistance to metronidazole can reach 95%. An inexpensive regimen is bismuth subsalicylate (two tablets) plus furazolidone (100 mg), four times daily for 4 weeks; however, as this yields a cure rate of only 72%, this regimen is not truly cost-effective. PMID- 9402351 TI - Rapid recurrence of Helicobacter pylori infection in Peruvian patients after successful eradication. Gastrointestinal Physiology Working Group of the Universidad Peruana Cayetano Heredia and The Johns Hopkins University. AB - Helicobacter pylori is associated with gastritis, peptic ulcer disease, and gastric cancer. Since gastric cancer is common in Peru, eradication of H. pylori may help to reduce the occurrence of gastric cancer. This study involved three randomized trials to determine the efficacy of four different triple-drug therapy regimens. The most successful regimen was furazolidone combined with bismuth subsalicylate and amoxicillin, which eradicated infection in 82% of patients. Patients successfully treated were followed every 2-3 months to determine the recurrence rate of H. pylori infection. Of 105 patients with H. pylori eradication documented by pathology and culture, 52% (55) returned for follow-up endoscopy, and in 73% (40) of these 55 the infection recurred during the 8-month follow-up period. Thirty-five patients from whom H. pylori was eradicated and who were tested for antibodies to H. pylori remained consistently seropositive. Rapid recurrence of H. pylori infection after successful eradication suggests that measures other than antimicrobial therapy are needed to fight H. pylori in developing countries. PMID- 9402353 TI - Value of serology as a noninvasive method for evaluating the efficacy of treatment of Helicobacter pylori infection. AB - The systemic humoral response to Helicobacter pylori was studied in 86 infected adult patients before antimicrobial therapy and at intervals following therapy. Endoscopy with collection of biopsy specimens was performed immediately before treatment; a 13C-labeled urea breath test was performed, and blood specimens were collected before treatment and at 1, 3, 6, 9, and 12 months after treatment. Serum samples from three patient groups (eradication success [n = 50], eradication failure [n = 16], and no treatment [n = 20]) were assayed for IgA and IgG antibodies to H. pylori by enzyme-linked immunosorbent assay. Levels of antibody to H. pylori before treatment were similar in all three groups. As expected, the no treatment and eradication failure groups had no significant changes in antibody levels during the study period. In contrast, for the eradication success group, the specific IgA and IgG antibody levels decreased progressively and significantly. We conclude that serology is a potentially useful way to monitor the success of treatment of H. pylori infection without using invasive or more expensive methods. PMID- 9402352 TI - Ranitidine versus colloidal bismuth subcitrate in combination with amoxicillin and metronidazole for eradicating Helicobacter pylori in patients with duodenal ulcer. AB - One hundred twenty-two patients were randomly assigned to three groups of treatment (A, B, and C), with (1) ranitidine (300 mg q.d. for 6 weeks), (2) ranitidine (300 mg q.d. for 6 weeks) with amoxicillin (500 mg t.i.d.) and metronidazole (500 mg b.i.d.) for the first 12 days, or (3) colloidal bismuth subcitrate (120 mg q.i.d. for 6 weeks) with amoxicillin and metronidazole (at same dosages as in the latter group). Six weeks after the beginning of treatment, an endoscopy showed that ulcers had healed in 49 of 52 patients (94.2%) from whom Helicobacter pylori had been eradicated and in 59 of 70 patients (84.3%) from whom it had not (NS). The rates of H. pylori eradication in groups A, B, and C were zero, 47.5%, and 86.8%, respectively. At 6, 12, and 18 months, an endoscopy was repeated for monitoring ulcer recurrence and H. pylori status. Reinfection rates at 6 months were 42.1% and 15.1% in groups B and C, respectively (P < .05). At 18 months, ulcers recurred in 82.9% (63) of 76 patients with noneradicated H. pylori infection, vs. 5.7% (2) of 35 patients without H. pylori infection (P < .001). We conclude that colloidal bismuth subcitrate is more effective for eradication of H. pylori than ranitidine when given with amoxicillin plus metronidazole for the treatment of duodenal ulcer, as both early reinfection and ulcer recurrence are diminished. PMID- 9402355 TI - Lactobacillus bacteremia and endocarditis: review of 45 cases. AB - Lactobacilli are part of normal gastrointestinal and genitourinary flora but are an uncommon cause of bacteremia. We reviewed the cases of 45 patients with clinically significant lactobacillus bacteremia occurring over 15 years. Underlying conditions were common, including cancer (40%), recent surgery (38%), and diabetes mellitus (27%). Twenty-two patients were in the intensive care unit at the time of onset of lactobacillus bacteremia. Eleven of the 45 patients were receiving immunosuppressive therapy, 11 were receiving total parenteral nutrition, and 23 had received antibiotics without activity against Lactobacillus prior to the occurrence of bacteremia. Bacteremia was polymicrobial in 27 patients and developed during hospitalization in 39. Thirty-one patients died, but only one death was attributable to lactobacillus bacteremia. Lactobacilli are relatively avirulent pathogens that produce bacteremia in patients with serious underlying illnesses, many of whom have received prior antibiotic therapy that may select out for the organism. While rarely fatal in itself, lactobacillus bacteremia identifies patients with serious and rapidly fatal illness. PMID- 9402354 TI - Acquired drug resistance in Mycobacterium tuberculosis isolates recovered from compliant patients with human immunodeficiency virus-associated tuberculosis. AB - We describe five compliant patients with human immunodeficiency virus (HIV) associated tuberculosis (TB) that relapsed, with acquisition of resistance by the original Mycobacterium tuberculosis strains. Both the first and second isolates from each patient had the same IS (insertion sequence) 6110-based DNA fingerprint patterns. Three of the five patients developed TB that was resistant to rifampin alone; no mutation in the region of the rpoB gene was detected by a line probe assay in two of the isolates from these patients. We discuss several factors presumably associated with acquired drug resistance in HIV-infected patients, including exogenous reinfection, drug interactions, malabsorption of drugs, and the presence of a large organism burden. PMID- 9402356 TI - Clostridium difficile infection is a risk factor for bacteremia due to vancomycin resistant enterococci (VRE) in VRE-colonized patients with acute leukemia. AB - A cohort study was conducted in a cancer center to identify risk factors for bacteremia with vancomycin-resistant enterococci (VRE) in neutropenic cancer patients colonized with VRE. There were 10 patients with VRE bacteremia among 56 colonized with VRE, of whose charts 51 were available for review. One hundred percent of patients with VRE bacteremia (10 of 10) vs. 56% of patients without VRE bacteremia (23 of 41) had acute leukemia (P = .01, Fisher's exact test). Four of the 10 patients with VRE bacteremia had a positive Clostridium difficile toxin assay within 6 days of their first positive VRE blood culture. Both C. difficile infection and antimicrobial (vancomycin and ciprofloxacin) use during VRE colonization were significant risk factors for VRE bacteremia in univariate analysis. When a Cox proportional hazards model was used to account for differences in follow-up time, C. difficile infection was the only statistically significant risk factor (risk ratio, 8.2; P = .007) for VRE bacteremia in VRE colonized patients with acute leukemia. PMID- 9402357 TI - Postsurgical mediastinitis: a case-control study. AB - We report the results of a case-control study of postsurgical mediastinitis (PSM) that we conducted from 1985 to 1993. The incidence of PSM was 2.2% (81 of 3,711 cases who underwent sternotomy); we analyzed the findings for 73 cases and 73 controls. Univariate analysis revealed that the risk factors for PSM were emergency surgery (27% of cases vs. 13% of controls), New York Heart Association functional class IV (46.5% vs. 21.9%), heart transplantation (12% vs. 0), and coronary artery bypass graft (CABG) surgery (60% vs. 41%). The incidences of fever, reoperation for bleeding, pacemaker placement, use of vasoactive drugs, prolonged mechanical ventilation, use of central lines, and treatment in the intensive care unit were also higher for cases. Multivariate analysis identified the following independent risk factors for PSM: reoperation (risk ratio [RR], 9.2), need for vasoactive drugs (RR, 3.5), CABG surgery (RR, 3.2), and fever that persisted after the third postsurgical day (RR, 406). The related mortality was 13.7%, and death was significantly more frequent among cases (17.7%) than among controls (2.7%). Multivariate analysis identified the following independent risk factors for mortality: bacteremia (RR, 21.5), the use of an intraaortic balloon (RR, 14.9), advanced age (RR, 1.14 per year), and prolonged mechanical ventilation (RR, 1.1 per day). PMID- 9402358 TI - Subcutaneous phaeohyphomycosis. PMID- 9402359 TI - Differential quantitative blood cultures in the diagnosis of catheter-related sepsis in intensive care units. AB - The aim of this prospective study was to compare differential blood cultures and quantitative catheter tip cultures for the diagnosis of catheter-related sepsis. Over a period of 2 years, 283 central venous catheters were inserted in 190 adult patients. Catheters were removed when they were no longer needed or when infection was suspected. Immediately before removal of the central venous catheters, blood cultures were performed, with blood drawn simultaneously from the catheter and the peripheral vein. After removal, quantitative catheter culture was performed according to the Brun-Buisson modified Cleri technique. Fifty-five quantitative catheter cultures were positive. They were classified as contaminated (n = 18), colonized (n = 23), or infected (n = 14). Differential blood cultures correctly identified 13 infections. With a catheter/peripheral cfu ratio of 8, differential blood cultures had a sensitivity of 92.8% and a specificity of 98.8%. When the catheters were removed because of suspected infection, differential blood cultures had a sensitivity of 92.8% and a specificity of 100%. Differential blood culture, a technique that does not necessitate catheter removal, seems effective in the diagnosis of catheter related sepsis in patients in the intensive care unit. PMID- 9402360 TI - A molecular epidemiological approach to studying the transmission of tuberculosis in Amsterdam. AB - We conducted a retrospective, population-based study with use of restriction fragment length polymorphism (RFLP) analysis to determine the incidence of and risk factors for clustering of Mycobacterium tuberculosis isolates, indicative of recently transmitted infection, among patients with culture-proven tuberculosis diagnosed between 1 July 1992 and 1 January 1995 in Amsterdam. We found that 214 (47%) of 459 patients were in 53 clusters, probably because of recent transmission of M. tuberculosis among 161 (35%) of these patients. Conventional contact tracing resulted in identification of 5.6% of the 161 patients. Clustering was more frequent among Dutch patients (59.3%) than among foreign ethnic patients (42.1%) (P = .002). The independent risk factor for clustering among Dutch patients was younger age; the independent risk factors among foreign ethnic patients were hard-drug use; alcohol abuse; and country of origin (Surinam or the Netherlands Antilles). These findings suggest the shortcomings of the usual tuberculosis control policies in Amsterdam. We identified several risk factors for clustering, which may guide adjustment of tuberculosis control and contact tracing strategies. PMID- 9402362 TI - Impaired pulmonary function in patients with hemorrhagic fever with renal syndrome. AB - Pulmonary and cardiac functions were investigated in 13 patients hospitalized with nephropathia epidemica, a European form of hemorrhagic fever with renal syndrome. As compared with reference values, the patients' diffusion capacity for carbon monoxide was decreased (P = .002) and pulmonary clearance of inhaled technetium-99m-labeled diethylenetriamine pentaacetic acid was increased (P = .002). In four of 11 patients, arterial blood gas analysis disclosed a reduction in partial pressure of O2 (< 10 kPa) and oxygen saturation (< 94%). In three of 13 patients, chest radiography revealed interstitial infiltrates or pleural effusions. Lung volumes and expiratory flow rates of the patients were not significantly changed. By electrocardiography and echocardiography, no significant cardiac dysfunction was demonstrable. The pulmonary dysfunction was best explained by an alveolocapillary lesion. The two hantavirus-caused clinical syndromes, hemorrhagic fever with renal syndrome and hantavirus pulmonary syndrome, may be pathophysiologically more similar than appears from the clinical presentations. PMID- 9402361 TI - Cytomegalovirus infection after intestinal transplantation in children. AB - Sixteen episodes of cytomegalovirus (CMV) disease occurred in 10 of 41 children undergoing intestinal transplantation from 1990 to 1995. Stratification of CMV disease by donor (D)/recipient (R) serological status was as follows: 3 of 8, D+/R-; 3 of 9, D+/R+; 4 of 9, D-/R+; and 0 of 15, D-/R-. Treatment resulted in resolution of CMV disease in 93.3% of episodes. No deaths attributable to CMV disease occurred in this series. CMV in D+/R- children resulted in more extensive and persistent disease. However, patient and graft survival rates were similar in the different D/R subgroups and between children with and without CMV disease. Cumulative dose of steroid boluses (relative risk [RR], 1.59; 95% confidence interval [CI], 1.14-2.21) and history of steroid recycles (RR, 2.72; 95% CI, 1.21 6.13) were associated with CMV disease. These results suggest that although CMV associated morbidity in pediatric intestinal transplant recipients was substantial, it was not associated with an increased rate of mortality or graft loss, even among high-risk D+/R- patients. PMID- 9402363 TI - Chlamydia pneumoniae in children with otitis media. AB - In this study the polymerase chain reaction was used to test for the presence of Chlamydia pneumoniae DNA in 118 middle-ear aspirates from 20 children with acute otitis media (AOM) and 53 children with otitis media with effusion (OME). C. pneumoniae was detected in 8 samples obtained from 5 children with OME and, together with Streptococcus pneumoniae, in a sample from 1 child with AOM. The mean age of these five children (6.6 +/- 1.4 years) was significantly higher than that of the 48 children with OME in whom C. pneumoniae could not be detected (4.3 +/- 1.9 years). The presence of C. pneumoniae in 9.4% of the examined children with OME suggests that C. pneumoniae might be a significant supplementary factor in the etiology of this common children's disease. PMID- 9402364 TI - Imipenem-resistant Pseudomonas aeruginosa: risk factors and antibiotic susceptibility patterns. AB - Potential risk factors for the detection of imipenem-resistant Pseudomonas aeruginosa in hospitalized patients were assessed by a case-control study. Forty patients whose first P. aeruginosa isolate was resistant or intermediate to imipenem were more likely than 387 controls to have received imipenem (odds ratio [OR] = 16.9; P < .0001) and to have undergone organ transplantation (OR = 3.9; P = .008). No significant difference was found for treatments with other antibiotics, other underlying diseases, demographic characteristics, different exposures to the hospital environment, or the culture site. Imipenem-resistant P. aeruginosa isolates were more likely to be resistant to other common antipseudomonal agents than were imipenem-susceptible isolates. It is concluded that treatment with imipenem, but not with other beta-lactam drugs, is a major risk factor for the detection of imipenem-resistant P. aeruginosa in hospitalized patients, that these organisms may relatively often be resistant to other antipseudomonal agents, and that the hospital environment per se might not play a major role in their epidemiology. PMID- 9402365 TI - Evaluation of pertussis in U.S. Marine Corps trainees. AB - One hundred twenty male U.S. Marine Corps trainees with histories of at least 7 days of cough underwent evaluation for Bordetella pertussis infection by culture, B. pertussis-specific polymerase chain reaction (PCR) analysis, and serology. Antibody levels in preexposure, acute-phase, and convalescent-phase serum samples were measured in a microagglutination assay and in enzyme linked immunosorbent assays (ELISAs) for IgG and IgA antibodies to pertussis toxin, filamentous hemagglutinin, pertactin, and fimbriae types 2 and 3. Culture and PCR analysis revealed that none of the patients were positive for B. pertussis; however, 20 of 120 trainees had serological evidence of B. pertussis infection. Of these cases, one was confirmed by a rise in the level of antibody to pertussis toxin, and six were classified as probable by increases in levels of antibodies measured by two or more assays. Of the 20 individuals with serological evidence of infection, 16 had rises in levels of antibodies to fimbriae or agglutinating antibodies. The utility of ELISA for detecting antibodies to fimbriae and the microagglutination assay for diagnosing pertussis in adults should be evaluated by application to larger and more diverse study populations. These results indicate that pertussis should be considered in the diagnosis of coughing illness in military populations. PMID- 9402366 TI - Pyogenic brain abscess caused by Streptococcus pneumoniae: case report and review. AB - While Streptococcus pneumoniae is the most common cause of bacterial meningitis in adults, cases of pneumococcal brain abscess have rarely been reported. We describe a case of otogenic brain abscess caused by S. pneumoniae that developed in a patient who was receiving ciprofloxacin for the empirical treatment of otitis media. We also review 23 additional cases of pyogenic brain abscess caused by S. pneumoniae that have previously been reported. The development of a pneumococcal brain abscess was associated with a contiguous intracranial focus of infection in 50% of cases. The majority of patients presented with headache (81%) and focal neurological deficits (86%). However, the classic triad of headache, fever, and focal neurological deficits was present in only 24% of patients. The mortality rate for patients with brain abscess caused by S. pneumoniae was 35%; persistent neurological deficits were documented in 40% of patients who survived. PMID- 9402367 TI - Duration of nasopharyngeal carriage of penicillin-resistant Streptococcus pneumoniae: experiences from the South Swedish Pneumococcal Intervention Project. AB - As a part of an intervention project, all detected carriers of penicillin resistant pneumococci (PRP) (MIC, > or = 0.5 mg/L) in Malmohus County, southern Sweden, were followed by means of weekly nasopharyngeal cultures. The median duration of carriage in 678 individuals was 19 days (range, 3-267 days). The duration of carriage was longest in children < 1 year old (median, 30 days) and shortest in adults > 18 years old (median, 14 days). Index cases, whose cultures were performed during an acute infection, were carriers for a mean of 10 days longer than asymptomatic contact cases (P < .05). The PRP spontaneously disappeared from the nasopharynx within 4 weeks in 68%, within 8 weeks in 87%, and within 12 weeks in 94% of the individuals. Other significant risk factors for prolonged carriage were the occurrence of > 6 episodes of acute otitis media (AOM) or first episode of AOM before the age of 1 year (P < .01), the carriage of PRP by other family members (P < .05), and the obtainment of a first positive culture during the winter months (P < .05). PMID- 9402368 TI - Nontyphoidal salmonella intracranial infections in HIV-infected patients. AB - Salmonella focal intracranial infections are unusual in human immunodeficiency virus (HIV)-infected patients. Six such infections have been reported in the world literature. We report a case of salmonella subdural and epidural cerebral empyema with concomitant osteomyelitis of the frontal bone. Such a complication in the course of salmonellosis is reported for the first time. In previously published case reports, four patients had brain abscess and two had subdural empyema. Salmonella typhimurium was isolated from two patients, and different serotypes were recovered from the others. All patients had advanced HIV disease, and all but two had had opportunistic infections before the diagnosis of salmonella intracranial infection. Surgical drainage combined with systemic antibiotic therapy resulted in the recovery of four of five patients. No regression of the lesions occurred in one patient treated only with antibiotics for multiple cerebral abscesses. PMID- 9402369 TI - Increased risk of maternal-infant hepatitis C virus transmission for women coinfected with human immunodeficiency virus type 1. Italian Study Group for HCV Infection in Children. AB - To estimate the risk of mother-to-child transmission of hepatitis C virus (HCV) and identify correlates of transmission, 245 perinatally exposed singleton children followed prospectively beyond 18 months of age were studied. Overall, 28 (11.4%) of the 245 children acquired HCV infection. Transmission occurred in 3 of 80 children (3.7%) whose mothers had HCV infection alone and in 25 of 165 (15.1%; P < .01) whose mothers had concurrent infection with human immunodeficiency virus type 1 (HIV-1). The percentage of HIV-1-infected children was similar (22 of 165, 13.3%), but each virus was transmitted independently; only six infants (3.6%) were coinfected with HCV and HIV-1. The risk of HCV transmission was not associated with maternal HIV-1-related symptoms, intravenous drug use, prematurity, low birth weight, or breast-feeding, whereas it was lower with cesarean section than with vaginal delivery (5.6% vs. 13.9%, P = .06). This suggests that transmission occurs mainly around the time of delivery. PMID- 9402370 TI - Effect of treatment with zidovudine on subsequent incidence of Kaposi's sarcoma. AB - Despite much investigation of zidovudine, little has been reported regarding its effect on the development of most individual AIDS-defining illnesses, including Kaposi's sarcoma (KS). We used observational data from the Multicenter AIDS Cohort Study (MACS) to estimate the effect of zidovudine use on the subsequent incidence of KS. To do this, we examined and adjusted for predictors of zidovudine use. CD4 lymphocyte counts, the development of HIV-related symptoms and AIDS, and changes in these factors were important predictors of zidovudine use. We used these associations to control for confounding by these and other factors with the G-estimation approach. We found no evidence that zidovudine use affected the time to KS in the MACS; the point estimate (95% confidence interval [CI]) for increase in time to KS was zero (-28%-68%). The relative risk was 1.0 (95% CI, 0.54-1.84). Randomized trials suggest that zidovudine may prevent KS. We discuss possible explanations for differences between results. PMID- 9402371 TI - Elevated concentrations of human neutrophil peptides in plasma, blood, and body fluids from patients with infections. AB - Neutrophil peptides, also called defensins, are antimicrobial molecules localized in the azurophil granules of neutrophils. We used a sensitive radioimmunoassay to measure the concentrations of human neutrophil peptides (HNPs) 1-3 in the plasma and blood of 86 healthy volunteers who served as controls and 54 patients with various infections. The respective mean plasma concentrations of HNPs 1-3 in the patients at the onset of bacterial infection, nonbacterial infection, and pulmonary tuberculosis were 4.2, 3.2, and 1.8 times the mean value (+/- SE) for the controls (254.8 +/- 7.1 pg/microL). Plasma concentrations of HNPs 1-3 for the patients were correlated with the peripheral blood neutrophil counts (correlation coefficient = 0.629; P = .0001). The mean concentration (+/- SE) of blood HNPs 1 3 in patients with bacterial infections was higher than the mean value (+/- SE) for controls (10.4 +/- 0.6 ng/microL), whereas the mean concentration (+/- SE) in patients with nonbacterial infections or tuberculosis did not differ from that for controls. Reverse-phase high performance liquid chromatography of plasma samples from patients with bacterial infections showed high concentrations of two precursors of HNPs 1-3, indicating that the biosynthesis of HNPs in neutrophil precursor cells is stimulated in response to infection and/or that the release of pro-HNPs is enhanced. HNPs 1-3 concentrations in the pleural fluid, bronchoalveolar lavage fluid, urine, and cerebrospinal fluid were also elevated in patients with infections. Our results suggest that HNPs have physiological significance in infection and that this family of peptides may be useful in assessing neutrophil function during infection. PMID- 9402372 TI - Questions and answers about defensins. PMID- 9402373 TI - Imported yellow fever in a United States citizen. AB - The last imported case of yellow fever seen in this country was in 1924. We report a case of yellow fever acquired by an American tourist who visited the jungles of Brazil along the Rio Negro and Amazon Rivers. The patient died 6 days after hospital admission and 10 days after his first symptoms appeared. Yellow fever virus was recovered from clinical specimens, and the isolate was genetically similar to the E genotype IIB of South American yellow fever viruses. This patient's illness represents a case of vaccine-preventable death since he failed to be immunized with a recommended preexposure yellow fever vaccine. PMID- 9402374 TI - Two strategies for managing invasive aspergillosis: a decision analysis. AB - We devised a diagnostic approach based on screening plasma for an Aspergillus antigen with use of a sandwich enzyme-linked immunosorbent assay (ELISA), thoracic computed tomographic scanning, and radionuclide imaging for managing patients at risk for invasive aspergillosis. We used a decision analytic model to compare this alternative strategy with the conventional strategy, which relies only on the presence of clinical symptoms, persistent fever, and chest roentgenographic findings. Use of the alternative strategy reduced the number of patients who would receive antifungal treatment empirically, but this strategy was more expensive. The specificity of the sandwich ELISA had a significant impact on cost, but the sensitivity did not. A 13% prevalence of infection resulted in equal costs for both strategies. As much as 43.3% of the patients treated empirically could be given liposomal amphotericin B (L-AmB) before the conventional strategy became the most expensive. The costs of the alternative strategy were less than those of the conventional strategy when >5.3% of all patients, irrespective of strategy, were treated with L-AmB. PMID- 9402375 TI - Spinal epidural abscess associated with the use of temporary epidural catheters: report of two cases and review. AB - Spinal epidural abscess has rarely been associated with the use of epidural catheters. We describe two patients with epidural abscesses that occurred in relation to the use of temporary epidural catheters; a literature review yielded 20 additional well-described cases. The mean age of these 22 patients was 49.9 years, the median duration of epidural catheter use was 3 days, and the median time to onset of clinical symptoms after catheter placement was 5 days. The majority of patients (63.6%) had major neurological deficits, and 22.7% also had concomitant meningitis. Staphylococcus aureus was the predominant pathogen. Despite antibiotic therapy and drainage procedures, 38% of the patients continued to have neurological deficits. These unusual but serious complications of temporary epidural catheter use require efficient and accurate diagnostic evaluation, as they can be substantial. PMID- 9402376 TI - Role of hemophagocytic histiocytosis in the etiology of thrombocytopenia in patients with sepsis syndrome or septic shock. AB - We conducted a study to assess the incidence and outcome of hemophagocytic histiocytosis (HH) in thrombocytopenic patients with sepsis syndrome or septic shock and to define the possible associations between HH, disseminated intravascular coagulation (DIC), and platelet-associated IgG (PAIgG) in promoting thrombocytopenia. Twenty immunocompetent thrombocytopenic patients were included. Bone marrow aspirates were obtained from each patient to identify hemophagocytic histiocytes. Coagulation parameters, PAIgG levels, and bacterial and viral infections were studied. Twelve patients with HH were identified. The presence of DIC and of PAIgG were often associated with this disease. No herpesvirus infection was demonstrated. Eight of the 12 patients with HH and four of the eight patients without HH died (P = NS). The results of this study suggest that HH could be involved in the development of thrombocytopenia in immunocompetent patients with sepsis syndrome or septic shock. HH does not seem to be associated with increased mortality. PMID- 9402377 TI - Streptococcus pneumoniae blood culture isolates from patients with and without human immunodeficiency virus infection: alterations in penicillin susceptibilities and in serogroups or serotypes. AB - We performed a 3-year retrospective study of Streptococcus pneumoniae blood culture isolates recovered at Baragwanath Hospital, Soweto, South Africa, from 1993 to 1995. The study group comprised 457 patients, including 98 children, of known human immunodeficiency virus (HIV) serostatus. Of these patients, 70 (30 [8.4%] of 359 adults and 40 [40.8%] of the 98 children) were infected with penicillin-resistant S. pneumoniae strains (minimal inhibitory concentration, > or = 0.12 microg/mL); 56 of these strains were intermediately resistant to penicillin. HIV-positive patients had significantly more penicillin-resistant isolates than did HIV-negative patients (43 [29.7%] of 145 HIV-positive patients vs. 27 [8.6%] of 312 HIV-negative patients; P < .001); this difference was found for both adults (19% vs. 4.3%; P < .001) and children (53.3% vs. 30.2%; P < .0343). Multiple resistance occurred more frequently in HIV-positive children (P = .02). HIV-positive adults had a statistically significant increase in the percentage of serogroups and serotype usually found in children and commonly associated with antimicrobial resistance, i.e., serotype 14 and serogroups 6, 19, and 23 (48% vs. 28.6%; P < .001). The increased prevalence of serogroups or serotypes usually found in children was also found among penicillin-susceptible strains. These data suggest that HIV-infected adults may again become susceptible to the serogroups or serotypes found in children. PMID- 9402379 TI - Group B streptococcal meningitis in adults: report of twelve cases and review. AB - Group B streptococcus (GBS) is the leading etiologic agent of bacterial meningitis and sepsis during the neonatal period, but it is an infrequent cause of meningitis in adults. We report 12 episodes of group B streptococcal meningitis in adults and review 52 cases reported in the literature. A total of 24 men and 40 women were included in the study; the mean age (+/- SD) was 49.2 +/ 20.5 years (range, 17-89 years). All the patients had cerebrospinal fluid cultures positive for GBS. Eighty-six percent of the patients had comorbid conditions, 50% had a distant focus of infection, and blood cultures yielded GBS for 78.7%. The overall case-fatality rate was 34.4% (22 patients). Factors associated with a poor outcome were advanced mean age (+/- SD) (61.5 +/- 17.4 years vs. 42.8 +/- 19.2 years; P = .0003) and the presence of complications on admission (P = .0001). Seven percent of survivors had neurological sequelae. Group B streptococcal meningitis in adults has become increasingly frequent in recent years; it tends to occur in patients with severe underlying conditions and is associated with a high case-fatality rate. Factors associated with a poor prognosis are advanced age and the occurrence of neurological and extraneurological complications. PMID- 9402378 TI - Randomized comparison of ganciclovir plus intravenous immune globulin (IVIG) with IVIG alone for prevention of primary cytomegalovirus disease in children receiving liver transplants. AB - A randomized placebo-controlled trial was conducted to determine the benefit of ganciclovir (5 mg/[kg x d]) for 30 days in addition to intravenous immune globulin (IVIG) for 16 weeks for prevention of primary cytomegalovirus (CMV) disease in children receiving liver transplants. Patients were monitored for 6 months after transplantation. The two groups of patients (recipients of 29 ganciclovir plus IVIG and 27 recipients of IVIG alone) were similar in terms of age, sex, and underlying disease. The incidence of CMV disease among the ganciclovir plus IVIG recipients and the IVIG alone recipients was 17% and 26%, respectively, and the time to disease in these recipients was 46 days and 32 days, respectively. There was no difference between groups in terms of survival; episodes of rejection, bacteremia, or fungemia; use of immunosuppressive agents; and incidence of leukopenia or thrombocytopenia. These results suggest that a 4 week course of ganciclovir with IVIG is not more effective than IVIG alone for prevention of primary CMV disease. Since short-term prophylaxis with ganciclovir may delay the onset of CMV disease, further studies with a longer course of ganciclovir prophylaxis are warranted. PMID- 9402380 TI - Efficacy of brompheniramine maleate for the treatment of rhinovirus colds. AB - We tested the efficacy of brompheniramine maleate in a large randomized, controlled trial of volunteers with experimental rhinovirus colds. Brompheniramine (12 mg) or placebo was administered at 8:00 A.M. and 8:00 P.M. for < or = 4 days after the onset of symptoms (24, 36, or 48 hours after virus challenge). During the first 3 days of treatment (the first 4 days after virus challenge), nasal secretion weights were lower for infected evaluable subjects receiving brompheniramine (n = 113) than for controls (day 1: 4.3 g vs. 6.8 g; day 2: 4.8 g vs. 7.7 g; and day 3: 3.3 g vs. 5.3 g) (P < or = .03), as were rhinorrhea scores (day 1: 0.6 vs. 0.8; day 2: 0.5 vs. 0.8; and day 3: 0.3 vs. 0.5) (P < .03), sneeze counts (day 1: 1.8 vs. 3.6; day 2: 2.1 vs. 5.1; and day 3: 1.3 vs. 3.3) (P < or = .001), and sneeze severity scores (day 1: 0.3 vs. 0.6; day 2: 0.25 vs. 0.7; and day 3: 0.2 vs. 0.4) (P < .001) (n = 112). Cough counts were lower after day 1 of treatment for the brompheniramine group than for controls (4.7 vs. 7.9) (P = .05) (day 2 after virus challenge), and other symptoms were modestly reduced or were unaffected in the brompheniramine group. Total symptom scores were also lower for the brompheniramine group than for controls on treatment days 1 (4.8 vs. 6.0) (P = .03) and 2 (4.1 vs. 5.6) (days 2 and 3 after virus challenge) (P = .003). Treatment with brompheniramine was associated with the adverse effects of somnolence (n = 3) and confusion (n = 1). Brompheniramine was efficacious treatment for the sneezing, rhinorrhea, and cough associated with rhinovirus colds. PMID- 9402381 TI - Quinolone arthropathy in animals versus children. AB - The use of quinolones in children and accumulation of data on the pharmacodynamics of these drugs have been limited and delayed by concern regarding their chondrotoxicity. A comprehensive review of the findings in animals compared with the cumulative published findings in children and adolescents (>7,000 to date) allows the conclusion that such concern is not justified. Prospective controlled studies in children are justifiable in view of a continuing lack of correlation between findings in juvenile animals and those in children and because of the selected therapeutic advantages of the current and newer quinolones. PMID- 9402382 TI - Prediction of relapse after treatment of coccidioidomycosis. AB - Relapse after apparently successful treatment of coccidioidomycosis has been a problem with both amphotericin B and the azoles. We conducted a retrospective cohort study of 34 patients who required therapy for coccidioidomycosis between 1973 and 1993; 10 relapsed and 25 (one patient received two courses of therapy) did not relapse during follow-up. The mean time to relapse after completion of therapy was 7.3 months (range, 1-21 months). All 34 patients responded clinically to therapy. A fourfold or greater decrease in titers of antibody, as determined by complement fixation (CF), during therapy was seen in seven (78%) of nine patients who relapsed and 17 (85%) of 20 patients who did not relapse (P = .956). There was no significant difference between relapsers and nonrelapsers in terms of the lowest CF titer during therapy, the CF titer at the end of therapy, or the peak CF titer. The risk of relapse was increased among those with a peak CF titer of > or = 1:256 (relative risk [RR] = 4.7; 95% confidence interval [CI] = 1.4 16.1), as compared with patients who did not mount such a high antibody response. Similarly, the risk of relapse was higher among those with serially negative coccidioidin skin tests (CSTs) than those with serially positive CSTs (RR = 4.8; 95% CI = 1.2-19.5). We conclude that clinical response, lowest CF titer, end-of therapy CF titer, and decrease in the CF titer of at least fourfold are not predictive of relapse in patients with coccidioidomycosis. Negative serial coccidioidin skin tests and a peak CF antibody titer of > or = 1:256 are independently associated with increased risk of relapse. PMID- 9402383 TI - Adequacy of therapy for coccidioidomycosis. PMID- 9402384 TI - The clinical use of fluoroquinolones for the treatment of mycobacterial diseases. AB - Mycobacterial diseases often require prolonged therapy with multidrug regimens. Fluoroquinolones have excellent bactericidal activity against many mycobacteria; achieve effective serum, tissue, and intracellular levels following oral administration; and produce few adverse effects. These properties have led to the increasing use of fluoroquinolones for the treatment of mycobacterial infections. We reviewed clinical studies and reports involving the use of fluoroquinolones for mycobacterial diseases. Ofloxacin, ciprofloxacin, sparfloxacin, and pefloxacin exhibit clinical efficacy against mycobacterial diseases, especially tuberculosis and leprosy. Fluoroquinolones have generally been administered in regimens that include other agents. However, when a fluoroquinolone has been found to be the sole active agent in a multidrug regimen, the ready emergence of resistance to fluoroquinolones has been recognized, just as when they have been used as monotherapy. Therefore, to forestall the emergence of resistance to fluoroquinolones during the treatment of mycobacterial diseases, these drugs should always be used in combination with at least one other active agent, and they should be used only when effective alternative drugs are not available. PMID- 9402385 TI - Acremonium species: new emerging fungal opportunists--in vitro antifungal susceptibilities and review. AB - We provide an overview of opportunistic fungal infections caused by Acremonium (Cephalosporium) species and discuss the classification of these species as well as the diagnosis and treatment of acremonium infections. We used a microdilution broth method to compare in vitro susceptibilities and minimum inhibitory concentrations and minimum fungicidal concentrations of amphotericin B, miconazole, itraconazole, 5-fluorocytosine, fluconazole, and ketoconazole for 33 clinical and environmental isolates of Acremonium. In general, the isolates tested displayed little susceptibility to the antifungals tested. Fluconazole and 5-fluorocytosine were ineffective in all cases. The efficacy of the remaining drugs was dependent on the strain. Amphotericin B showed the best results. PMID- 9402386 TI - Nosocomial infections in human immunodeficiency virus-infected patients in a long term-care setting. AB - To our knowledge, the epidemiology of hospital-acquired infections in human immunodeficiency virus (HIV)-infected patients during long-term care has not been reported. For 13 months, we observed HIV-infected patients (50 men and 15 women) in a dedicated 21-bed unit in a long-term-care facility to determine the rate of nosocomial infections. The mean age of the patients was 39 years (range, 22-78 years); 74% of the patients had CD4 cell counts of < 200/mm3. There was a total of 152 infections (24 infections per 1,000 long-term-care days). The factors associated with the occurrence of a nosocomial infection were low CD4 cell counts, poor functional status, and longer duration of stays at the facility. The three most common infections were Clostridium difficile-associated diarrhea, primary bacteremia, and urinary tract infection. Eighteen hospital-manifested opportunistic infections occurred. More than 50% of the cases of bacteremia were due to multidrug-resistant organisms. Nosocomial infections occur commonly in HIV infected patients in long-term care and thus are important considerations in patient management. PMID- 9402387 TI - Endogenous interleukin-2 serum levels in children infected with human immunodeficiency virus. AB - Levels of interleukin-2 (IL-2) in serum obtained from human immunodeficiency virus (HIV)-infected children at health maintenance visits were measured to characterize endogenous IL-2 responses and to examine the association between these responses and progression of immunosuppression. IL-2 was detectable (level >8.7 pg/mL) in the serum of 28 of 45 HIV-infected children; 42% (19 of 45) had serum IL-2 levels of >39 pg/mL. Children without evidence of immunosuppression (Centers for Disease Control and Prevention Pediatric HIV Classification Immunologic Category 1, n = 15) and children with severe immunosuppression (immunologic category 3, n = 20) had statistically significant lower serum IL-2 levels (mean +/- [SD], 134.4 +/- 227.3 pg/mL and 18.2 +/- 30.3 pg/mL, respectively) than those with moderate immunosuppression (mean +/- [SD], 450.5 +/ 311.8 pg/ml; immunologic category 2, n = 10) (P < .05, Wilcoxon rank sum test). In those children in whom immunosuppression was evident, decreasing serum IL-2 levels correlated with depletion of CD4+ lymphocytes (r = 0.74), whereas there was an inverse correlation between serum IL-2 levels and CD4+ lymphocyte counts (r = -0.47) in children with no or moderate immunosuppression. PMID- 9402388 TI - Acute Epstein-Barr virus infection complicated by severe thrombocytopenia. AB - We describe one patient with acute Epstein-Barr virus (EBV) infection associated with severe thrombocytopenia and review 36 additional cases reported in the literature. Complications of EBV infection due to severe thrombocytopenia occurred in 10 (27.0%) of 37 patients, and 2 (5.4%) of 37 patients died. Although acute EBV infections are generally benign and self-limiting, thrombocytopenia, a potentially serious complication, should not be overlooked. PMID- 9402390 TI - Rhabdomyolysis associated with acute Q fever. PMID- 9402389 TI - Effect of eliminating seropositive canines on the transmission of visceral leishmaniasis in Brazil. AB - In Brazil, where Leishmania chagasi causes endemic American visceral leishmaniasis (AVL), the spread and maintenance of human disease are attributed to canine reservoirs. However, despite measures directed toward the elimination of infected canines, the incidence of human disease continues to increase. To evaluate the role of infected canines in the acquisition of AVL by humans, we undertook a controlled intervention study in three similar, but isolated, valleys of Pancas, Espirito Santo, Brazil. In the two experimental (intervention) valleys, infected dogs were eliminated whereas in the control valley, seropositive canines remained untouched. During the 12-month study period, human seropositivity rates, as measured by dot enzyme-linked immunosorbent assay, increased from 15% to 54% in the intervention valleys and from 14% to 54% in the control valley. The elimination of infected canines in the intervention valleys did not result in a statistically significant difference between the incidences of human serological conversion in the intervention and control valleys at either 6 (20% and 22%, respectively; P = .5961) or 12 months (26% and 27%, respectively; P = .9442). The role of humans as a significant reservoir for AVL is proposed as an explanation for the study results. PMID- 9402391 TI - Hepatic abscess: rare complication of ventriculoperitoneal shunts. PMID- 9402392 TI - Mycobacterium lentiflavum: an etiologic agent of cervical lymphadenitis. PMID- 9402393 TI - Adoptive immunotherapy for interstitial pneumonia associated with cytomegalovirus infection. PMID- 9402394 TI - Isolation of toxigenic Clostridium difficile from dialysate fluid in a fatal case of chronic ambulatory peritoneal dialysis-related peritonitis. PMID- 9402395 TI - Postpartum epidural abscess due to group B Streptococcus. PMID- 9402396 TI - Recovery of small colony variants of Staphylococcus aureus following gentamicin bead placement for osteomyelitis. PMID- 9402397 TI - Phaeoacremonium parasiticum infective endocarditis following liver transplantation. PMID- 9402398 TI - Invasive cryptococcosis in a family with epidermodysplasia verruciformis and idiopathic CD4 cell depletion. PMID- 9402399 TI - IgG response to pneumococcal polysaccharide-protein conjugate appears similar to IgG response to polysaccharide in bone marrow transplant recipients and healthy adults. PMID- 9402400 TI - Ocular toxoplasmosis after autologous peripheral-blood stem-cell transplantation. PMID- 9402401 TI - Salmonella neck abscesses. PMID- 9402402 TI - Treatment of acyclovir-resistant herpes simplex virus keratitis in a patient with Wiskott-Aldrich syndrome. PMID- 9402403 TI - Coinfection with human immunodeficiency virus and human T-cell lymphotropic virus type I: reciprocal activation with clinical and immunologic consequences. PMID- 9402404 TI - Pulmonary coccidioidomycosis in Japan: case report and review. PMID- 9402405 TI - Hemorrhagic dengue with spontaneous splenic rupture: case report and review. PMID- 9402406 TI - High serum ferritin concentration in an AIDS patient with miliary tuberculosis. PMID- 9402407 TI - Bacteremia due to Burkholderia gladioli: case report. PMID- 9402408 TI - Cutaneous protothecosis in a patient with AIDS and a severe functional neutrophil defect: successful therapy with amphotericin B. PMID- 9402409 TI - Fatal acute hepatic necrosis due to dose-dependent fluconazole hepatotoxicity. PMID- 9402410 TI - Fever, erythroderma, abdominal pain, and renal failure following initiation of indinavir therapy. PMID- 9402411 TI - High-dose ampicillin plus streptomycin for treatment of a patient with severe infection due to multiresistant enterococci. PMID- 9402412 TI - Renal dysfunction in a human immunodeficiency virus-infected patient who was treated with indinavir. PMID- 9402413 TI - Nonsteroidal antiinflammatory drugs: concurrent or causative drugs in serious infection? PMID- 9402415 TI - The role of glucose-6-phosphate dehydrogenase deficiency in blackwater fever. PMID- 9402414 TI - Transmission of tuberculosis during medical procedures. PMID- 9402416 TI - High levels of adenosine deaminase in patients with aseptic meningitis. PMID- 9402417 TI - [Chronic depression in the year 2000: therapeutic hopes. 10th World Congress for Psychiatry. Madrid, 24 August 1996]. PMID- 9402418 TI - [Herpes genitalis, an underrated disease?]. PMID- 9402419 TI - [Amiodaron improves prognosis in high risk patients]. PMID- 9402420 TI - [Mibefradil: a new class calcium inhibitor]. PMID- 9402421 TI - [ACE-inhibitors indicated even in non-diabetic nephropathy]. PMID- 9402422 TI - [Uniform guidelines for manuscripts for publication in biomedical periodicals 1997. Abbreviations for titles in periodicals. International Committee of Medical Journal Editors]. PMID- 9402423 TI - [The Giessen glaucoma symposium. Glaucoma: new aspects of research and therapy]. PMID- 9402424 TI - [New therapeutic methods in the treatment of multiple sclerosis]. PMID- 9402425 TI - [New methods in the pharmacotherapy of Parkinson disease]. PMID- 9402426 TI - [Effective anxiolytic therapy with Buspirone. Focusing on modulation of the serotonin-neurotransmitter system]. PMID- 9402427 TI - The relationship between electromyography and work intensity revisited: a brief review with references to lacticacidosis and hyperammonia. AB - The aim of this investigation was to re-evaluate the relationship between electromyography and work intensity during incremental work in light of highly discrepant literature. Trained male subjects participated in the study (n = 14). Each subject completed a VO2max test on a cycle ergometer. Tests started at a power output of 60 Watts with a 30 Watt.4 min-1 work increment. Each test was terminated at exhaustion. Blood was collected at the end of each work intensity for lactate and ammonia analysis. EMG were recorded from the vastus lateralis, rectus femoris and vastus medialis using pre-amplified surface electrodes. EMG were collected at each intensity over a period of 60 cycle revolutions. EMG signals were analyzed using integration and EMG spectral analysis. Gas exchange variables were recorded on-line for each test (15 second interval). Ammonia and lactate threshold points were surpassed at the same absolute work intensity (200 Watts) which was equivalent to 64-69% VO2max. When a linear model was applied to the iEMG data, coefficients of determination achieved were greater than those obtained when an exponential model was used for the vastus lateralis and medialis. Gradients of regression lines fitted to iEMG data at pre- and post lactate/ammonia threshold work intensities were not different. Alternatively, the iEMG-work intensity relationship for the rectus femoris muscle tended to be curvilinear. Significant increases in iEMG were observed at post-lactate/ammonia threshold work intensities for the rectus femoris reflecting increases in fatigue and type II motor unit recruitment at these intensities. In general, median frequency of the EMG power spectrum function were unchanged during incremental work, although highly individualistic results were observed between some subjects and muscles. Grouped median frequency values were insensitive to changes in recruitment, metabolite accumulation and fatigue associated with the increases in work intensity. Consequently, the usefulness of EMG spectral analysis during incremental work was questioned. PMID- 9402428 TI - Early detection of diabetic neuropathy: a neurophysiological study on 100 patients. AB - As a long-term complication of diabetes mellitus, autonomic neuropathy has received considerable attention in the last few years since a tripled 5-year mortality of patients with diabetic neuropathy including autonomic disturbances has been observed. In a total of 100 diabetics with clinical manifest neuropathy of different stages (N0 = 8, N1 = 15, N2a = 24, N2b = 28, N3 = 25), 26 of whom were insulin-dependent and 74 non-insulin-dependent, the validity of different neurophysiologic and autonomic function test procedures proposed in literature was assessed. Apart from clinical examination, nerve conduction velocity measurement of five nerves as well as amplitude measurement of evoked sensory and motor actin potentials, electromyography of at least four muscles of the lower limbs, and measurement of sympathetic skin response on hands and feet were performed. Furthermore, heart rate variation at deep periodical breathing (E/I ratio), during Valsalva's manoeuver and after standing up (30/15-ratio) was determined. In addition, the drop in systolic blood pressure to standing up was measured. The further developed the clinical picture of neuropathy, the more pathological were the results obtained in different tests. The results suggest that most changes leading to pathological values of nerve conduction velocity and heart rate variation measurement take place in a clinical stage, in which no or only very slight clinical signs give evidence of diabetic neuropathy (N0-N1). Therefore, especially these examinations should be performed on diabetics with no or only slight clinical signs of neuropathy in order to reveal those with beginning neuropathic disturbances. EMG examination is preferable in later stages of the disease. PMID- 9402429 TI - Auditory event-related potentials in Parkinson's disease. AB - We studied 49 patients with Parkinson's disease (PD) by a neuropsychological battery examining the temporo-spatial orientation, short-term memory, comprehension, non-verbal intelligence, long-term memory and anomia and the Auditory Event-Related Potentials. In the patients the latencies of the N100 and N200 waves were prolonged and the amplitude of the P300 wave was reduced compared with controls. No difference was found in the ERP of patients with and without cognitive deficits. Equally, no correlation was found between the ERP, the cognitive impairment, the length or the severity of the disease evaluated by Hoehn-Yahr's and Webster's scales. PMID- 9402430 TI - Short and long latency cortical potentials evoked by electrical stimulation of the oesophageal mucosa in normal alert humans. AB - Cerebral responses from the oesophagus were investigated in 16 normal male and female volunteers ranging in age from 20 to 54 years. The stimulus was applied by a naso-oesophageal probe equipped with bipolar ring electrodes. Short and long latency EP (SLEP and LLEP) were observed in all the subjects examined. SLEP consisted in a low threshold potential of 30 to 70 microV amplitude, biphasic or triphasic in shape and of approximately 5 to 10 ms duration; mean latency at the largest peak was 4.5 +/- 1.7 at 25 cm from the nostrils. Early components at about 2.5-3.5 ms and of small amplitude are also present. Recording from the neck at C7 with a common non-cephalic reference, SLEP components occurred from 2 to 6 ms earlier than that from the scalp, suggesting an oligo-synaptic transmission of the excitement via ganglion and lemniscal pathways to the cortex. SLEP was always followed by a complex potential formed of a succession of negative and positive waves with latencies ranging from 20 to 300 ms: the LLEP. This LLEP was usually not very stable and reproducible during the course of successive recordings and in the same subject because it tended to adjust. Preliminary observations concerning the topographical cortical distribution of oesophageal evoked potentials show a circumscribed localization of the SLEP in the parieto-temporal region of the hemisphere whereas LLEP was more widespread. It is the authors' opinion that oesophageal evoked potentials are generated both by the excitation of myelinic fibres with a wide range of conduction speed and of amyelinic fibres from the oesophageal mucosa and the paraoesophageal peripheral nerves of vagal origin. PMID- 9402431 TI - Brainstem auditory evoked potentials (BAEPs) and somatosensory evoked potentials (SEPs) in enteric encephalopathy (EE). AB - BAEPs and SEPs were studied in 25 patients of enteric encephalopathy in acute phase and the results were compared with 25 healthy control persons. In the study the important observations of BAEPs were delayed peak latency of wave III, wave V and delayed ILP I-V, and of SEPs was prolonged peak latency of N20. The electrophysiological evidence suggests metabolic cause for the coma and the SEP changes were similar to those observed in cerebral malaria reported earlier in this laboratory. PMID- 9402432 TI - Three-Hertz postural oscillation in patients with brain stem or cerebellar lesions. AB - Postural instability was quantitatively studied in patients with lesions in the 1) pontine tegmentum, 2) deep cerebellar nuclei, or 3) cerebellar hemisphere. As compared to controls, patients with each lesion demonstrated a characteristic 3 Hertz (Hz) body oscillation. Cross-correlation functions revealed a strong correlation between the body sway and activities in the lower leg muscles. Based on these findings, we conclude that the 3 Hz body oscillation may be the result of any disturbance in the loop of long-latency reflexes mediated by the cerebellum. PMID- 9402433 TI - H-reflex latency: a maturity criterion for newborn babies. AB - One hundred newborn babies were investigated for H-reflex latency (H-RL) in between 33 to 40 weeks (wk) of post-conceptional age (PCA). Weekly data of both preterms (33 to 36 wk PCA) and fullterms (37 to 40 wk PCA) were compared. A significant reduction in H-RL was noticed at 37 wk PCA when newborns attain their term. The babies could be classified into preterms and fullterms by using their H RL values. PMID- 9402434 TI - Comparison of surface EMG signals between electrode types, interelectrode distances and electrode orientations in isometric exercise of the erector spinae muscle. AB - The influence of electrode type, interelectrode distance (IED) and electrode orientation on EMG signals from the paraspinal muscles was investigated. Bipolar electrodes were placed at distances 2, 3, 4, 6 and 8 cm over the erector spinae in the cranio-caudal direction ("in series") as well as in the direction perpendicular to it ("in parallel"). Ten subjects performed 5 s isometric contractions of the erector spinae at 20, 40, 60, 80 and 100% MVC by pulling upward on a handlebar attached to the floor. RMS EMG signals were analyzed for mean average amplitude (AA). Mean total power (TP) and mean median frequency (MF) of the raw EMG signal were determined using fast Fourier transform. In addition to graded loading, sustained fatiguing contractions were performed from which TP and MF were obtained. With increasing IED the AA and TP increased while MF decreased. Although a trend towards higher AA, TP and MF was found for electrodes "in series", as compared to those "in parallel", the difference never reached significance. It is concluded that consistent information about muscle activity was obtained with Miniature Biopotential Skin Electrodes and 14445C Hewlett Packard electrodes independently from IED or orientation. Orientation "in parallel" prevented the electrodes from sliding during muscle contraction. The third tested type, electrodes developed in the Neuromuscular Research Center, Boston, proved extremely sensitive to movement. PMID- 9402435 TI - Optimizing cholesterol lowering therapy: contribution of the Post Coronary Artery Bypass Graft Trial. PMID- 9402436 TI - Digoxin in heart failure: results of the recent Digoxin Investigation Group trial in the context of other treatments for heart failure. PMID- 9402437 TI - Percutaneous mitral commissurotomy: an effective treatment in 'ideal' candidates whatever the approach. PMID- 9402438 TI - The absence of ventricular premature beats on a Holter is like a normal sedimentation rate. PMID- 9402439 TI - Determinants of peak aerobic capacity after heart transplantation. PMID- 9402440 TI - Low energy intracardiac cardioversion of atrial fibrillation. PMID- 9402441 TI - Cholesterol lowering in coronary disease: a step in the right direction. PMID- 9402442 TI - Gap junctions and clinical cardiology: from molecular biology to molecular medicine. PMID- 9402444 TI - Endothelial dysfunction and accelerated coronary artery disease in cardiac transplant recipients. Microcirculation Working Group, European Society of Cardiology. PMID- 9402443 TI - Reperfusion strategies in acute myocardial infarction. PMID- 9402445 TI - Compliance and adverse event withdrawal: their impact on the West of Scotland Coronary Prevention Study. AB - AIMS: To assess the additional benefit gained from high compliance in the West of Scotland Coronary Prevention Study and to examine cases where withdrawal from trial medication was due to an adverse event. METHODS: The incidence of definite coronary heart disease or non-fatal myocardial infarction, cardiovascular mortality, definite or suspect coronary heart disease death or non-fatal myocardial infarction, the need for coronary revascularization procedures, all cause mortality and incident cancers were measured in the entire cohort and compared with the high compliance group. The adverse events associated with withdrawal were coded by body system. RESULTS: In subjects with compliance > or = 75%, treatment with pravastatin resulted in a 38% risk reduction for definite coronary heart disease death or non-fatal myocardial infarction and for cardiovascular mortality, a 46% reduction in risk or coronary revascularization and a 32% risk reduction (P = 0.015) for all-cause mortality. CONCLUSIONS: The analysis of the effect of pravastatin in the subgroup of high compliers to randomized medication demonstrated a substantial increase in the estimated risk reductions in comparison with that achieved in the intention-to-treat analysis. This result has significant implications for the motivation of high compliance among patients and for the assessment of the cost-effectiveness of treatment. PMID- 9402446 TI - Cholesterol lowering after participation in the Scandinavian Simvastatin Survival Study (4S) in Finland. AB - BACKGROUND: Patient compliance is crucial for the effectiveness of preventive medication. The aim of the study was to investigate changes in serum cholesterol levels and the use of cholesterol lowering drugs one year after the end of the Scandinavian Simvastatin Survival Study (4S), a randomized secondary prevention study of coronary heart disease with simvastatin and placebo. METHODS AND RESULTS: A questionnaire asking the current use of cholesterol lowering drugs, most recent serum cholesterol value and attitudes towards cholesterol lowering was sent to 785 surviving 4S participants in four 4S centres in Finland. The response rate was 94%. The current use of cholesterol lowering drugs and the reported mean serum cholesterol values were similar to the original simvastatin and placebo groups. In all, 74% (n = 546) reported that they had used cholesterol lowering drugs after the study, and 63% (n = 467) were currently using them, mostly simvastatin (96%) with an average dose of 14 (SD 5) mg.day-1. Cholesterol lowering was considered to be 'very important' by 53% and 'important' by 37% of the respondents. The most frequent reasons for discontinuation were 'drug costs' (38%) and 'normal cholesterol values' (30%). The reported mean serum cholesterol levels were 5.1 (SD 1.0) and 5.7 (SD 1.1) mmol-1 in the current cholesterol lowering drug users and non-users, respectively (P < 0.0001). The in-trial treatment goal of serum cholesterol (< or = 5.2 mmol-1) was not met in 38% of the users and in 68% of the non-users of cholesterol lowering drugs. CONCLUSION: One year post-trial the original simvastatin and placebo groups of the 4S had become similar with regard to the use of cholesterol lowering drugs and serum cholesterol levels. The adherence to medication, however, still remained relatively high, but there was a shift toward lower doses, and consequently toward higher post-trial serum cholesterol levels. PMID- 9402447 TI - Prevalence of coronary artery disease and coronary risk factors in rural and urban populations of north India. AB - OBJECTIVE: This study was conducted to determine and compare the prevalence of coronary artery disease and coronary risk factors in both a rural and an urban population of Moradabad in north India. DESIGN AND SETTING: A cross-sectional survey of two randomly selected villages from the Moradabad district and 20 randomly selected streets in the city of Moradabad. SUBJECTS AND METHODS: The 3575 subjects were between 25 and 64 years old; 1769 (894 men and 875 women) lived in the countryside and 1806 (904 men and 902 women) lived in the city. The survey methods were questionnaires, physical examination and electrocardiography. RESULTS: The overall prevalence of coronary artery disease, based on a clinical diagnosis and an electrocardiogram, was 9.0% in the urban and 3.3% in the rural population. The prevalences were significantly (P < 0.001) higher in the men compared with the women in both urban (11.0 vs 6.9%) and rural (3.9 vs 2.6%) populations, respectively. The prevalence of symptomatic coronary artery disease (known coronary disease and Rose questionnaire-positive angina) was 2.3% in the men (n = 19) and 1.5% in the women (n = 13) in the rural subjects, and 8.5% in the men (n = 77) and 3.4% in the women (n = 31) in the urban population. When diagnosed on the basis of electrocardiographic changes alone, the prevalences were 1.5% (n = 26) in the rural population and 3.0% (n = 55) in the urban. Coronary risk factors were two- or three-fold more common among urban subjects compared to the rural population in both sexes. Central obesity was four times more common in the urban population compared to the rural in both sexes. Sedentary lifestyle and alcohol intake were significantly (P < 0.01) higher in the urban population compared to the rural subjects. There was a significant association between coronary disease and age, hypercholesterolaemia, hypertension and central obesity in both sexes. Smoking was a significant risk factor of coronary disease in men. CONCLUSIONS: Coronary artery disease and coronary risk factors were two or three times higher among the urban compared with the rural subjects, which may be due to greater sedentary behaviour and alcohol intake among urbans. It is possible that some Indian populations can benefit by reducing serum cholesterol, blood pressure and central obesity and increasing physical activity. PMID- 9402448 TI - 'Expected infarct size without thrombolysis', a concept that predicts immediate and long-term benefit from thrombolysis for evolving myocardial infarction. AB - BACKGROUND: Thrombolytic therapy should only be used when expected benefits outweigh the risks. In order to obtain a precise estimation of prognosis, with and without thrombolytic therapy, we postulated that mortality reduction by thrombolytic therapy is a function of the area of myocardium at risk for necrosis. We developed a model to estimate the myocardial area at risk for necrosis from clinical parameters readily available upon hospital admission. This model was validated in relation to long-term prognosis and benefits of thrombolytic therapy. METHODS: Enzymatic infarct size with and without thrombolysis was predicted from the haemodynamic state and the electrocardiogram on hospital admission by multivariate regression analysis in 885 patients in the rt-PA placebo and rt-PA/PTCA trial of the European Cooperative Study Group. This multivariate function was used to validate the 'expected infarct size without thrombolytic treatment' in a test population of 533 patients from the Intracoronary Streptokinase trial of the Interuniversity Cardiology Institute of The Netherlands (ICIN) and 1741 patients from the Intravenous Streptokinase in Acute Myocardial Infarction (ISAM) study, both trials with a non-thrombolysed control group. RESULTS: Expected infarct size correlated well with the actual enzymatic infarct size in the non-thrombolysed patients of the latter two series. Limitation of infarct size by thrombolytic therapy was greatest in patients with a large 'expected infarct size' and absent in patients with a small area at risk. Similarly, one year mortality reduction was greatest in patients with a large 'expected infarct size without thrombolysis'; four deaths were prevented per hundred (95% confidence interval 0 to 9) if the area at risk was large, vs one death (95% confidence interval -2 to 3) in patients with a small area at risk. Benefit was most pronounced in patients with a large area at risk who were treated early within 3 h of symptom onset. A score for the determination of 1 year mortality with and without thrombolytic therapy is presented to help the clinician determine who to treat with thrombolytic therapy. CONCLUSION: 'Expected infarct size without thrombolysis' is a useful tool for clinicians to estimate the amount of myocardium at risk of necrosis in individual patients and to decide whether thrombolytic therapy is warranted. It is the only validated parameter of myocardium at risk for necrosis that is readily available for all patients with myocardial infarction and does not need high-tech equipment. PMID- 9402449 TI - Right coronary artery stenosis is associated with impaired cardiac endocrine function during exercise. AB - AIMS: Resting plasma levels of atrial natriuretic peptide and B-type natriuretic peptide rise with left ventricular dysfunction, but little is known about effects of cardiac ischaemia on atrial natriuretic peptide and B-type natriuretic peptide levels during exercise. We investigated exercise levels of atrial natriuretic peptide and B-type natriuretic peptide in patients with suspected angina to determine whether these measurements could improve non-invasive assessment of coronary disease severity. METHODS AND RESULTS: One hundred patients performed an exercise test (Bruce protocol) within 2 weeks of coronary angiography. Plasma levels of atrial natriuretic peptide and B-type natriuretic peptide were measured at rest and at peak exercise. Multivariate regression analysis was used to assess effects of age, sex, coronary anatomy, exercise time and ventricular function on atrial natriuretic peptide and B-type natriuretic peptide levels. Increasing age and female sex were significantly associated with higher resting atrial natriuretic peptide levels; age alone was associated with higher exercise atrial natriuretic peptide levels. As expected, left ventricular end-diastolic pressure and disease of left anterior descending and circumflex coronary arteries were associated with increased resting B-type natriuretic peptide levels. However, the usual rise in B-type natriuretic peptide levels during exercise was independently reduced by disease of the right coronary artery. CONCLUSION: This paradoxical effect of right coronary artery disease limits the value of natriuretic peptide measurements as predictors of coronary disease severity. Impaired release of B type natriuretic peptide may reduce exercise tolerance in patients with right coronary artery disease. PMID- 9402450 TI - Addition of felodipine to metoprolol vs replacement of metoprolol by felodipine in patients with angina pectoris despite adequate beta-blockade. Results of the Felodipine ER and Metoprolol CR in Angina (FEMINA) Study. Working Group on Cardiovascular Research, The Netherlands (WCN). AB - AIMS: The study aimed to compare the addition of felodipine to metoprolol, and of the replacement of metoprolol by felodipine, with continuation of metoprolol, in patients with angina pectoris despite optimal beta-blockade. METHODS AND RESULTS: The study was double-blind, parallel, randomized and controlled, and comprised 363 patients from 27 outpatient cardiology clinics in the Netherlands. The patients had angina and positive bicycle exercise tests despite optimal beta blockade (resting heart rate < 65 beats.min-1). Randomization was to three treatment groups: continuation of metoprolol (control), addition of felodipine to metoprolol, and replacement of metoprolol by felodipine. Exercise tests were repeated after 2 and 5 weeks. The main outcome measure was: exercise result after 5 weeks, compared with baseline, between-group comparison of changes vs control. There were no significant differences in exercise duration and onset of chest pain vs control. The addition of felodipine increased time until 1 mm ST depression (43 s, 95% confidence interval 20-65 s), and decreased both ST depression at highest comparable work load (0.46 mm, 95% confidence interval 0.19 0.72), and maximal ST depression (0.49 mm, 95% confidence interval 0.23-0.74). Exercise results after replacement of metoprolol by felodipine were not different from control, apart from a significant increase in rate pressure product. Significantly more patients experienced adverse events in the felodipine monotherapy group. CONCLUSION: Combination of metoprolol and felodipine is to be preferred to felodipine monotherapy in patients who have signs and symptoms of myocardial ischaemia despite optimal beta-blockade. PMID- 9402452 TI - Trends in hospitalization and mortality for heart failure in Spain, 1980-1993. AB - AIMS: To describe, for the first time, trends in hospitalization and mortality rates for congestive heart failure in Spain during the period 1980-1993. METHODS AND RESULTS: Data on primary diagnosis of congestive heart failure were taken from the National Hospital Morbidity Survey and the National Vital Statistics. The number of hospital admissions for congestive heart failure rose by 71% (from 42,965 in 1980 to 73,448 in 1993) and hospitalization rates for congestive heart failure increased by 47% (from 348 per 100,000 in 1980 to 511 per 100,000 in 1993). The rise in hospitalizations was limited to persons aged > or = 65 years, and proved greater among women. Congestive heart failure was the leading cause of hospitalization in persons aged > or = 65 years, accounting for 5% of all hospital admissions in this age group. Age-adjusted congestive heart failure mortality declined by 23%. The decline affected all age groups, with the sole exception of the > or = 80-year group in which mortality rose. Nevertheless, congestive heart failure remained the third leading cause of cardiovascular death. CONCLUSION: Congestive heart failure represents a significant hospital and demographic burden for the Spanish population. The hospital burden increased substantially in the period 1980-1993, and will continue to do so in future with the growth of the elderly population. PMID- 9402451 TI - Balloon mitral valvotomy: comparison between antegrade Inoue and retrograde non transseptal techniques. AB - AIMS: The results of percutaneous mitral valvotomy performed by the antegrade transseptal method using the Inoue balloon (n = 1000; group 1) and by the retrograde non-transseptal technique using a polyethylene balloon (n = 100; group 2) were compared in a retrospective, non-randomized study. METHODS AND RESULTS: Both the groups were similar with respect to baseline characteristics. The success rate was 95% in group 1 and 93% in group 2. There was a significant increase in mitral valve area estimated by Gorlin's equation (Group 1: from 0.8 +/- 0.5 to 2.1 +/- 0.8 cm2; Group 2: from 0.8 +/- 0.3 to 1.9 +/- 0.8 cm2, both P < 0.001) and by Doppler echocardiography using the pressure half-time method (Group 1: from 0.9 +/- 0.4 to 2.2 +/- 0.6 cm2; Group 2: from 0.9 +/- 0.3 to 2.0 +/- 0.7 cm2, both P < 0.001). However, the calculated immediate post-valvotomy mitral valve area was larger with the Inoue technique (2.1 +/- 0.8 vs 1.9 +/- 0.8 cm2; (P < 0.02). Results were considered optimal when the mitral valve area increased to > or = 1.5 cm2, the percentage increase was > or = 50, and mitral regurgitation was < or = 2/4. Out of the total successful procedures, optimal results were obtained in 95% patients in Group 1 and 94% in Group 2. Incidence of significant mitral regurgitation (> or = grade 3/4) was similar in two groups (Group 1: 4% vs Group 2: 5%, P = ns). A significant left to right atrial shunt (Qp/Qs > or = 1.5:1) in 2.5% and tamponade in 2% of cases occurred exclusively with the Inoue technique, while conduction disturbances, such as transient (< 24 h) left bundle branch block (28%) and complete heart block (2%) were noted with the retrograde technique (Group 2). Local complications were significantly higher in Group 2 (3% vs 0.5%, P < 0.01). The procedure time with the Inoue technique was shorter than with the retrograde (Group 1: 15 +/- 8, range 10 to 35 min; Group 2: 22 +/- 14, range 15 to 45 min, P = 0.05). Echocardiographic follow-up at 1 year showed no significant difference in mitral valve area between the two groups (Group 1 (n = 300): 1.8 +/- 0.8 vs Group 2 (n = 60): 1.9 +/- 0.9 cm2; P = 0.3). CONCLUSIONS: Balloon mitral valvotomy using the Inoue balloon and the retrograde non-transseptal technique results in significant immediate haemodynamic and symptomatic improvement. The Inoue technique achieved a larger immediate post-valvotomy mitral valve area, but the difference was not apparent at 1 year follow-up. Incidence of significant mitral regurgitation was similar with both the techniques; however, local complications occurred more frequently with the retrograde technique. Both techniques may complement each other in technically difficult cases. PMID- 9402453 TI - A prospective, randomized comparison of temperature-controlled vs manually delivered radiofrequency catheter ablation in patients undergoing atrioventricular nodal modification or accessory pathway ablation. AB - AIMS: In a prospective, randomized study, the effect of temperature control on radiofrequency catheter ablation was compared in 69 patients undergoing atrioventricular nodal modification (n = 32) or ablation of an accessory pathway (n = 37). METHODS AND RESULTS: Thirty-five patients were randomized to temperature control, 34 to manually delivered radiofrequency ablation. The success rate was 92.5% for accessory pathway ablation and 100% for atrioventricular nodal modification. Mapping duration was significantly reduced only in patients undergoing atrioventricular nodal modification. The number of applications was higher for manually delivered ablation in patients undergoing atrioventricular nodal modification (5.6 +/- 1.1 vs 1.9 +/- 0.4, P = 0.004) as was the cumulative energy delivered (5034 +/- 1008 vs 2054 +/- 517 W, P = 0.013) whereas the mean power per application was higher with temperature control (41.4 +/- 1.8 vs 34.1 +/- 1.1 W, P = 0.002). No significant differences in these parameters were found in patients undergoing accessory pathway ablation. Coagulum formation on the catheter tip was observed more often with manually delivered ablation 5.3% vs 0.9%, P = 0.026). The success rate with the initially randomized application mode was higher for temperature control (94.3 vs 61.8%, P = 0.003). CONCLUSIONS: Temperature control during radiofrequency current ablation significantly reduces mapping duration, necessary applications and cumulative energy in atrioventricular nodal modification, but not accessory pathway ablation. Coagulum formation on the catheter tip still occurs but is significantly reduced compared to manually delivered radiofrequency current. PMID- 9402454 TI - Survival and incidence of myocardial infarction in men with ambulatory ECG detected frequent and complex ventricular arrhythmias. 10 year follow-up of the 'Men born 1914' study in Malmo, Sweden. AB - AIM: To assess to what extent do frequent or complex ventricular arrhythmias, detected during 24 h ambulatory electrocardiographic recording (ECG), influence prognosis with regard to survival and incidence of ischaemic heart disease. METHODS AND RESULTS: The study subjects were the 456 randomly selected men born in 1914, the population-based cohort study of 1982-83, in Malmo, Sweden. The main outcome measures were total mortality and incidence of cardiac event (myocardial infarction and death from ischaemic heart disease). Frequent or complex ventricular arrhythmias (Lown classes 2-5) were detected in 49% of the men with (n = 77), and in 35% of those without, a history of myocardial infarction or angina pectoris at baseline, P = 0.019. Independent of clinically evident coronary artery disease at baseline, and after adjustment for traditional atherosclerotic risk factors and use of digitalis or beta-blocker therapy, frequent or complex ventricular arrhythmias were associated with an increased mortality from ischaemic heart disease (relative risk (RR), 2.1; 95% confidence interval (CI), 1.2-3.9) and an increased cardiac event rate (RR, 1.6; 95% CI, 1.0 2.5)). Men free from both ischaemic-type ST depression and frequent or complex ventricular arrhythmias (used as the control group) had the lowest ischaemic heart disease death rate, 5.9 per 1000 person-years. The combination of ST depression and frequent or complex ventricular arrhythmias was associated with an ischaemic heart disease death rate of 20.9 per 1000 person-years. The cardiac event rate in these two groups was 15.6 and 76.1 per 1000 person-years, respectively (adjusted RR, 2.3; CI, 1.1-4.6). CONCLUSIONS: In elderly men without a history of myocardial infarction and angina pectoris, frequent or complex ventricular arrhythmias during ambulatory ECG recording is associated with an increased incidence of myocardial infarction and mortality. Men who, during ambulatory ECG recording, also demonstrate ST-segment depression have an even less favourable prognosis. PMID- 9402455 TI - A comparison of treatment of atrial fibrillation with low-energy intracardiac cardioversion and conventional external cardioversion. AB - AIM: Low-energy (1 to 15 J), catheter-based intracardiac cardioversion was compared with transthoracic external cardioversion (360 J) in a prospective, cross-over clinical trial. METHODS AND RESULTS: In 187 consecutive patients with chronic atrial fibrillation, over a period of a mean of 10.0 +/- 7.3 (SD) months, 217 cardioversion attempts were made. Intracardiac shocks were randomly applied between two 6-F catheters located in either the right atrium and coronary sinus or between the right atrium and left pulmonary artery. When a cardioversion attempt with one method failed, the other method was implemented. After cardioversion, all patients were treated orally with sotalol with a mean daily dose of 174 +/- 54 mg. Internal cardioversion was more effective than external cardioversion (65/70 = 93% vs 92/117 = 79%, P < 0.01). The mean energy for successful cardioversion was 5.8 +/- 3.2 J for the internal and 313 +/- 71 J for the external cardioversion group. At a mean follow-up of 12.5 +/- 6.4 months, 48% (38%) of the patients treated with internal (external) cardioversion were in sinus rhythm (P < 0.05). In 22 of 25 patients in whom external cardioversion failed, sinus rhythm was restored with internal cardioversion at a mean energy of 6.5 +/- 3.0 J. Overweight patients had twice the risk of unsuccessful external cardioversion. CONCLUSIONS: Internal cardioversion is effective in restoring sinus rhythm. It might be indicated in patients in whom external cardioversion had failed or in whom external cardioversion is assumed to be difficult or even contraindicated. PMID- 9402456 TI - Electrophysiological properties in patients undergoing atrial compartment operation for chronic atrial fibrillation with mitral valve disease. AB - AIMS: Surgical treatment for atrial fibrillation is now feasible in selective cases. The aim of this study was to assess the electrophysiological properties of patients undergoing atrial compartment operation for chronic atrial fibrillation. METHODS AND RESULTS: Electrophysiological studies were performed in 20 mitral valve patients with atrial fibrillation who had been maintained in sinus rhythm for more than 1 year after atrial compartment operation. Intra-cardiac recording and programmed electrical stimulation were performed in various atrial compartments. The parameters studied included sinus node function, atrial conduction and refractoriness, atrioventricular conduction function and inducible arrhythmias if any. Intra-cardiac recordings showed that the rhythm was of sinus origin in all cases, with the earliest atrial activity located in the high right atrium. The mean sinus cycle length was 750 +/- 110 ms, AH time 106 +/- 29 ms, and HV time 53 +/- 7 ms. The sinus node function was normal in 18 patients (90%), and only two patients had prolonged sinus node recovery and sino-atrial conduction. The right atrial appendage compartment was driven by the sinus node in all patients. However, the conduction time from the high right atrium to the right atrial appendage compartment was markedly prolonged in 12 of 15 patients (80%) undergoing the three-compartment operation in which an incision was placed between the high right atrium and right atrial appendage compartments. On the other hand, the electrical activities in the left atrial compartment were much more varied. In 13 of 20 patients (65%), the left atrial compartment was driven by the sinus node; 11 of the 13 patients had a normal or mildly prolonged conduction time (ranged 75 to 146 ms), whereas two patients had a marked delay in conduction (200 ms and 266 ms, respectively). In the remaining seven patients, the left atrial compartments were dissociated from the rest of the heart; five of them had a quiescent left atrium, one a fluttering left atrial rhythm, and one a slow left atrial rhythm. The effective refractory period was longer in the left atrial compartment (242 +/- 47 ms) as compared to that of the high right atrium (224 +/- 26 ms, P < 0.01) and right atrial appendage compartments (219 +/- 25 ms, P < 0.01). Programmed electrical stimulation could not induce atrial fibrillation in any patient, whereas two patients had inducible atrial flutter and three repetitive atrial responses. CONCLUSIONS: (1) Atrial compartment operation does not impair sinus node function in most cases. (2) Elimination of atrial fibrillation while maintaining the electrical connection between different atrial compartments is feasible. PMID- 9402457 TI - Cardiopulmonary physiology after surgical closure of asymptomatic secundum atrial septal defects in childhood. Exercise performance is unaffected by age at repair. AB - AIMS: Most secundum atrial septal defects, once diagnosed, are corrected at a young age. The evidence to justify early vs delayed or even non-closure is equivocal and little is known regarding long-term effects of later closure. This is particularly pertinent to those patients awaiting transcatheter closure of their defect for whom a device is only just becoming available. We examined the exercise cardiorespiratory physiology of children surgically treated for an isolated secundum defect. METHODS AND RESULTS: One hundred and six healthy control children and 22 children more than 6 months after surgical repair for an isolated secundum atrial septal defect were studied. All were asymptomatic. Measurements of effective pulmonary blood flow, stroke volume, arteriovenous oxygen difference, minute ventilation, heart rate, oxygen consumption and carbon dioxide production were made using a quadrupole mass spectrometer during rest and graded exercise. Data from the normal children allowed calculation of z scores for the atrial septal defect group matched for age, sex, pubertal stage and surface area. Maximal exercise performance was equal between control and atrial septal defect groups, however, the atrial septal defect group had a significantly greater effective pulmonary blood flow and stroke volume but a lower heart rate than controls at a given exercise stage. Stroke volume abnormalities were most closely related to duration of follow-up (29% of the variance explained, P < 0.01) rather than age at surgery. CONCLUSIONS: We were unable to show a medium term benefit from early surgery for an asymptomatic secundum atrial septal defect during exercise. The clinical relevance of the haemodynamic differences that do exist remains unclear. PMID- 9402458 TI - Predictive factors of maximal aerobic capacity after cardiac transplantation. AB - Exercise capacity in cardiac transplanted patients has been reported to remain decreased in some studies; however, functional results after transplantation may vary, ranging from modest to spectacular improvement. The aim of the study was to quantify exercise capacity in a large series of transplanted patients and to search for factor predictive of a good functional result. Eighty-five patients (mean 52.1 +/- 11.8 years) underwent exercise testing with respiratory gas exchange measurements 1 to 100 months after transplantation. Mean performance was 112.4 +/- 33 W with a peak VO2 of 21.1 +/- 6 ml.min-1.kg-1. Heart rate was 103 +/ 14 at rest, reaching 142 +/- 22 beats.min-1 at the end of exercising. In univariate analysis, maximal or submaximal aerobic capacity parameters were strongly correlated with chronotropic reserve (r = 0.63; P < 0.001) without correlation with cold ischaemic time, number of rejection episodes or right bundle branch block. In multiple regression analysis, chronotropic reserve, time from transplantation, age of donor and age of patient were proved to be the variables best correlated with peak VO2. Our study confirms the persistence of a large decrease in aerobic functional capacity despite cardiac transplantation; limited exercise capacity does not improve over time, and is limited not only by the patient's age but by that of the donor, and especially by chronotropic reserve. PMID- 9402460 TI - C-reactive protein and complement in myocardial infarction and postinfarction heart failure. PMID- 9402459 TI - Catecholamines contribute to exertional dyspnoea and to the ventilatory response to exercise in normal humans. AB - BACKGROUND: Exogenous catecholamine administration in humans stimulates ventilation. The present study was designed to investigate whether increased endogenous catecholamine release influences objective measures of ventilation and subjective measures of breathlessness in normal subjects. METHODS: Yohimbine, a pre-synaptic alpha 2 adrenoceptor antagonist, or placebo was administered to 10 normal male subjects in a double-blind cross-over fashion. Ventilation and metabolic gas exchange were measured during steady state exercise at 60% of previously determined maximal oxygen consumption. Venous lactate and noradrenaline were measured during exercise. Subjects' sensation of breathlessness and fatigue were recorded using visual analogue scales. RESULTS: Plasma noradrenaline was higher following yohimbine administration (at 6 min exercise; 4.58 +/- 0.56 nmol.l-1 vs 8.74 +/- 1.53; P < 0.05). Oxygen consumption was unchanged, but ventilation was greater throughout exercise following yohimbine. The sensation of exertion was greater following yohimbine, and at any given level of ventilation, the sensation of exertion was greater. CONCLUSIONS: Yohimbine administration causes increased noradrenaline release. This is associated with an increased ventilatory response and an increase in the sensation of exertion during steady state exercise. An increase in circulating noradrenaline might be a mechanism for both increased ventilation and pathological conditions of breathlessness such as chronic heart failure. PMID- 9402461 TI - Aspirin therapy in diabetic subjects post-myocardial infarction. PMID- 9402462 TI - Valvular heart disease in systemic lupus erythematosus: another symptom of the disease or a hallmark of secondary antiphospholipid syndrome? PMID- 9402463 TI - Is thrombolytic therapy beneficial to acute stroke patients? PMID- 9402464 TI - Arterial structural modifications in hypertension. Effects of treatment. AB - Hypertensive changes in the vasculature occur at all levels of the circulation- from the large arteries through to the microcirculation. Detection of these changes may offer useful predictive information in assessing cardiovascular risk and the need for treatment. Recent evidence shows that at least some of these changes are reversible with anti-hypertensive treatment. PMID- 9402465 TI - Rationalizing the heart failure trials: from theory to practice. AB - It is 10 years since the CONSENSUS I study showed that ACE inhibitors improved mortality in heart failure. This finding has been confirmed in numerous trials, for example SOLVD, SAVE. Indeed, in the intervening 10 years, many other potential therapies have been examined in mortality trials, but so far no other therapy has had as good effect on mortality as ACE inhibitors. The other therapies which have been examined are digoxin, amlodipine, beta-blockers, amiodarone, etc. Despite ACE inhibitors being a very effective therapy for heart failure, there is still remarkable under-use of them in clinical practice. The reason for this needs to be explained further, but fear of hypothermia and renal dysfunction appear to be major factors. PMID- 9402466 TI - Adequate blood pressure control: perception and reality. AB - Blood pressure should be controlled over 24 h to reduce or prevent cardiac hypertrophy and reduce the prevalence of sudden death, myocardial infarction and myocardial ischaemia at the time of the morning rise in blood pressure. Anti hypertensive medication is usually given once-daily in the morning and the dose is titrated on the basis of post-dose (peak) blood pressure. This frequently leads to inadequate control prior to the next drug dose unless drugs with appropriate pharmacokinetics or pharmacodynamics are used. ACE inhibitors exhibit an E(max) plasma concentration: blood pressure response relationship, and thus short-acting ACE inhibitors can exert an effect over 24 h if titrated based on pre-dose (trough) blood pressure. However, when titrated in clinical practice on post-dose (peak) blood pressure response, doses are used that are inadequate to control blood pressure for 24 h. ACE inhibitors with appropriate pharmacokinetics such as perindopril can control blood pressure when titrated at peak provided a dose is used (4 or 8 mg) that is known to have a T:P close to 1.0. Shorter-acting ACE inhibitors frequently give inadequate control when titrated at peak. An understanding of the pharmacokinetics and pharmacodynamics of a drug, coupled with knowledge of the time the drug was taken, together with the time of blood pressure measurements, enables control to be achieved with once-daily therapy even if the drug is titrated at peak response. PMID- 9402467 TI - From kinetics to dynamics: are there differences between ACE inhibitors? AB - Angiotensin converting enzyme inhibitors are established treatment for hypertension and heart failure. There are well documented differences between ACE inhibitors both in physicochemical properties and pharmacokinetics. Pharmacodynamic actions are similar for most members of the ACE inhibitor class but there are compounds with additional effects which may reflect protease inhibition or non-enzyme-directed pharmacological properties. Clinically relevant differences are few and far between, particularly in the treatment of hypertension when the optimal dose and dose intervals are used. In heart failure there may be a role for drugs with additional properties such as neutral endopeptidase inhibition. In addition, ACE inhibitors differ in the profile of blood pressure changes after the first dose. Early haemodynamic changes with a fall in blood pressure in heart failure patients may be disadvantageous in terms of subsequent outcome. Thus the haemodynamic effects of the first dose may be relevant to the choice of ACE inhibitors in heart failure. PMID- 9402468 TI - Endothelial dysfunction and atherosclerosis. AB - The endothelium mediates a number of responses (relaxation or contraction) of arteries and veins from animals and humans. The endothelium-dependent relaxations are due to the release, by endothelial cells, of potent non-prostanoid vasodilator substances. Among these, the best characterized is endothelium derived relaxing factor (EDRF), which is believed to be nitric oxide (NO). Nitric oxide is formed by the metabolism of L-arginine by the constitutive NO synthase of endothelial cells. In arterial smooth muscle, the relaxation evoked by EDRF is explained by the stimulation by NO of soluble guanylate cyclase that leads to the accumulation of cGMP. In a number of animal blood vessels and in human coronary arteries, the endothelial cells release a substance that causes hyperpolarization of the cell membrane (endothelium-derived hyperpolarizing factor, EDHF). The release of EDRF from the endothelium can be mediated by both pertussis toxin sensitive (alpha 2-adrenoceptor activation, serotonin, aggregating platelets, leukotrienes) and insensitive (adenosine diphosphate (ADP), bradykinin) G proteins. In blood vessels from animals with regenerated and reperfused endothelium, and/or atherosclerosis, there is a selective loss of the pertussin toxin-sensitive mechanism of EDRF release, which favours the occurrence of vasospasm, thrombosis and cellular growth. The available information from isolated human blood vessels or obtained in situ concurs with the conclusions reached from studies with isolated animal tissues. In addition to relaxing factors, the endothelial cells can produce contracting factors (endothelium derived contracting factors; EDCFs) which include superoxide anions, endoperoxides, thromboxane A2 and endothelin. From animal studies it can be concluded that the propensity to release EDCFs is maintained, or even augmented, in diseased blood vessels. The switch from a normally predominant release of EDRFs to that of EDCFs may play a crucial role in atherosclerosis. PMID- 9402469 TI - The stimulatory effect of rabphilin 3a on regulated exocytosis from insulin secreting cells does not require an association-dissociation cycle with membranes mediated by Rab 3. AB - Rabphilin 3a is a Rab 3-GTP binding protein concentrated on secretory vesicles of neurons and endocrine cells. There is evidence that rabphilin 3a undergoes cycles of association-dissociation with membranes and that recruitment of rabphilin 3a to secretory vesicles is mediated by Rab 3a, suggesting that rabphilin 3a is a downstream effector of this Rab. In this study we have investigated whether a membrane-anchored form of rabphilin 3a mimics the action of rabphilin 3a on secretion and bypasses the need for Rab 3 function. Overexpression of both wild type rabphilin 3a and of a transmembrane anchored form of rabphilin 3a stimulated (about 2-fold) evoked secretion of coexpressed human proinsulin from clonal HIT T15 cells. A similar transmembrane-anchored protein which lacked the Rab 3 binding region stimulated secretion even more effectively. Unexpectedly, a rabphilin 3a deletion mutant missing the Rab 3 binding domain was also stimulatory on secretion, although a further deletion of rabphilin to exclude the first of the two proline-rich regions abolished its stimulatory effect. The first of these two mutants was primarily particulate, while the second mutant was primarily soluble, suggesting that the first proline-rich region of rabphilin 3a plays a role in targeting rabphilin to its site of action. We conclude that the action of rabphilin 3a can be independent of Rab 3 if other mechanisms produce a sufficient concentration of the protein in proximity of exocytotic sites. These results provide new evidence for a fundamental similarity in the mechanisms by which Ras and Rab GTPase produce their distinct physiological effects. PMID- 9402470 TI - Rab3 dissociation and clathrin-mediated endocytosis, two key steps in the exo endocytotic pathway of large dense-cored vesicles in primary cultures of superior cervical ganglia. AB - In this study we have used primary cultures of porcine superior cervical ganglia as a model system to study exo-endocytosis in sympathetic neurons. Pure neuronal cultures with a defined noradrenergic phenotype can be obtained when antimitotics are included in the culture medium, and the high yield from prenatal piglets allows a biochemical approach in addition to morphological studies. Release of large dense-cored vesicles (LDCVs) was visualized by exposing stimulated neurons to anti-dopamine-beta-hydroxylase (D beta H) prior to fixation. Double immunofluorescent staining revealed that synaptotagmin was colocalized with anti D beta H labeled exocytotic spots, but not the small GTP-binding protein rab3. Density gradient centrifugation of a postmitochondrial supernatant of cultured cells confirmed the dissociation of rab3 from a population of mature LDCVs. As expected, rab3 did not reassociate with the lighter D beta H-positive membrane fraction in the gradient representing internalized LDCV membranes. The fluid phase marker horseradish peroxidase colocalized with retrieved LDCV membranes. Indirect immunofluorescence demonstrated the colocalization of clathrin with D beta H-positive exocytotic spots on the plasma membrane. At the ultrastructural level, depolarization of neurons resulted in the abrupt loss of LDCVs and concomitant appearance of clathrin-coated vesicles and coated omega profiles. The size distribution of coated structures overlapped strongly with that of LDCVs. Taken together, these data clearly suggest that two key regulatory events in the process of exo-endocytosis, i.e. rab3 dissociation and clathrin-mediated internalization, are not only reminiscent of small synaptic vesicles, but also are a feature of the LDCV pathway at the presynaptic plasma membrane of sympathetic neurons. PMID- 9402471 TI - De novo acquisition of neuronal polarity in retinoic acid-induced embryonal carcinoma cells. AB - The mouse embryonal carcinoma cell line PCC7-Mz1 represents an advantageous model to study acquisition of polarity by neurons. During the first two days after differentiation is induced by the addition of retinoic acid, the neuronal derivatives develop extensions which for at least four more days do not differ from each other in growth characteristics, morphology, and marker expression. Beginning around differentiation day 6 and following the relocation of the nucleus from a central to a polar position in the cell soma, the morphology and marker expression changes dramatically: expression of MAP2 diminishes and eventually disappears in the thinner neurite (future axon), which originates at the nucleated pole, but remains strong in the branched, broad based neurite(s). The opposite changes in expression are observed for synaptophysin, together with a clustering of the vesicle protein in varicosity-like areas. Complete segregation of expression of the two markers is achieved around day 12, shortly followed by dendrite-specific location of MAP2 mRNA and the ability to generate and conduct action potentials. Our studies add several aspects to the process of neuronal polarity acquisition, as it was previously studied in primary cultures of embryonic neurons: (i) we monitored neuronal differentiation from the birth of neurons, rather than from later and less defined maturation stages, (ii) cell nucleus relocation may be associated with the induction of neuronal polarity, and (iii) functional competence of neurons is closely associated with previous acquisition of polarity. Acquisition of polarity by PCC7-Mz1 neuronal derivatives probably refers to de novo acquisition rather than to reestablishment of polarity. PMID- 9402472 TI - Triglyceride synthesis and secretion and lipogenesis implicated gene expression in the chicken hepatocarcinoma cell line LMH. AB - Avian lipogenesis was studied in the chicken hepatocarcinoma LMH cell line. The differentiated and lipogenic status of these cells was evidenced by the presence of the albumin mRNA as well as of some mRNA coding for enzymes involved in lipogenesis (acetyl-CoA carboxylase, fatty acid synthase, delta 9 desaturase) and for apoproteins (apoprotein B and A1). These results were further confirmed by the analysis of triglyceride synthesis and secretion rates in growing cells. A time course analysis showed that triglyceride metabolism was affected by cell density. Hormone responsiveness of triglyceride production was also analyzed. Insulin, triiodothyronine and glucagon to a lesser extent were shown to regulate lipogenesis of LMH cells. The results were compared with those obtained in primary cultures of chicken hepatocytes. PMID- 9402473 TI - Modulation of osteogenesis and adipogenesis by human serum in human bone marrow cultures. AB - Knowledge of the controlling mechanisms of human osteoprogenitor cell differentiation has important implications for understanding bone turnover. The in vitro differentiation of human bone marrow fibroblasts into adipogenic and osteogenic cells and the interaction of 1,25 dihydroxyvitamin D3 (1,25(OH)2D3) and dexamethasone in this process has been investigated together with the effects of human serum. Marrow fibroblasts cultured in human serum and dexamethasone for 28 days, generated lipid containing cells as confirmed by morphology, Oil red O staining and immunocytochemistry using antiserum to the adipocyte-specific protein, adipocyte P2 (aP2). In cultures containing 1,25(OH)2D3 and dexamethasone, adipogenesis was stimulated within 21 days. Osteocalcin expression, as assessed by in situ hybridization, was dependent on the presence of 1,25(OH)2D3 and was decreased in cultures treated with dexamethasone. Northern analysis confirmed the decrease in osteocalcin expression and increase in lipoprotein lipase expression with the appearance of the adipogenic phenotype in these cultures. Marrow cultures maintained for 14 days in human serum and osteotropic agents before switching to fetal calf serum indicated the continuous requirement of human serum in these cultures for adipogenesis. These results demonstrate that human serum contains factors that exert dramatic effects on human bone marrow cell differentiation to augment the osteogenic and adipogenic activity of 1,25(OH)2D3 and dexamethasone. PMID- 9402474 TI - Effect of PMA on the integrity of the membrane skeleton and morphology of epithelial MDCK cells is dependent on the activity of amiloride-sensitive ion transporters and membrane potential. AB - The signaling pathways from an activation of protein kinase C (PKC) by phorbol myristate acetate (PMA) to the rearrangement of actin-based cytoskeleton and membrane skeleton of epithelial MDCK cells were studied by visualizing the cytoskeletal organization with immunofluorescence microscopy and by measuring intracellular pH, sodium ion concentration and membrane potential with the aid of fluorescent intracellular indicators. Upon PMA treatment the MDCK cells lost their cubic shape and acquired a spindle-like morphology. The stress fibers were depolymerized, and fodrin, the main component of the membrane skeleton, was released from the lateral walls to the cytosol. These changes were accompanied by depolarization of the cells, decrease in the intracellular pH and sodium ion concentration. In order to test the mutual correlation between the PMA-induced alterations we treated the cells with PMA in the presence of channel inhibitors or ionophores and in defined media. The effects of PMA on the membrane skeleton and morphology could be reversed in media lacking Na+ or K+ ions or by hyperpolarizing agents, dimethylamiloride and valinomycin, suggesting that the effects of PMA on the cytoskeleton were dependent on the ion gradients and membrane potential across the cell membrane. Moreover, the morphological changes and instabilization of the membrane skeleton of MDCK cells took place spontaneously without PMA in depolarizing conditions, in potassium gluconate buffer. We suggest that the membrane potential across the cell membrane of MDCK cells together with the activity of amiloride-sensitive cation transporters transmits signals in the protein kinase C (PKC) pathway leading from activation of PKC to fibroblast-like morphology and cytoplasmic localization of membrane skeleton components, features characteristic for cancer cells. PMID- 9402475 TI - Deuterium oxide (heavy water) accelerates actin assembly in vitro and changes microfilament distribution in cultured cells. AB - While deuterium oxide (D2O) is known to produce various biological effects in living animals and cultured cells, the detailed mechanisms by which it does so remain unclear. The present study was designed to assess the effects of D2O on microfilaments (MFs) via fluorescence staining of BALB 3T3 cells and in vitro actin polymerization studies. After BALB 3T3 cells had been exposed to a concentration of more than 30% D2O for several hours, stress fibers in the peripheral region became thick and distinct, while the quantity of perinuclear MFs was drastically reduced. This effect was transient and returned to the original distribution within 12 h. Cytoplasmic F-actin (FA) also increased transiently coincident with the enhancement of stress fibers. The pattern of cell locomotion became simpler, and total locomotor activity was suppressed in a D2O concentration-dependent manner. Analysis of in vitro studies demonstrated that, when purified G-actin was polymerized in D2O at a concentration greater than 10%, the rate of actin polymerization was accelerated, whereas the total amount of polymerized actin at the steady state in D2O was the same as that in H2O controls. A gelation assay and transmission electron microscopy (TEM) showed that the network of crosslinked FA with alpha-actinin became denser in 30% D2O than in H2O. These findings concerning actin polymerization and FA gelation suggest that the alteration of stress fibers in cultured cells is caused by a direct effect of D2O on cellular MF dynamics. PMID- 9402476 TI - Copy number, epigenetic state and expression of the rRNA genes in young and senescent rat embryo fibroblasts. AB - The recent cloning of the gene that causes the premature aging in Werner syndrome patients has evoked speculations that deficits in expression of the ribosomal RNA genes could be related to cellular aging in general. Here we compare the state of the rRNA genes and the rRNA metabolism in young and senescent (aged) rat embryo fibroblasts (REF). Southern blot analysis revealed that the copy number and the methylation state of the genes did not change significantly with increasing cumulative population doublings (CPD) of the culture. Hence, young (low numbers of CPD) and senescent REF (high numbers of CPD), respectively, have the same repertoire of rDNA units that can be transcribed. The rRNA synthesis in these cells was analyzed by incorporation of labeled uridine at conditions allowing the measurement of absolute rather than relative rRNA synthesis rates. We revealed that the cell density dependence of the rRNA synthesis diminishes in senescent cells. Exponentially growing young REF exhibited an rRNA synthesis of 16 amol uridine incorporation per minute and cell. The rRNA synthesis decreased 10-fold in quiescent cells at saturating cell densities. Exponentially growing REF near the end of their replicative lifespan exhibited a 2-fold lower rRNA synthesis rate compared to young cells. However, in senescent REF the rRNA synthesis rate decreased only 2-fold with increasing cell densities resulting in a 3-fold higher rRNA synthesis rate compared to young cells at saturating cell densities. These data could be confirmed by calculating the rRNA synthesis rates from the rRNA content, the rRNA half-life, and the proliferation rate of the cells. Hence, senescent REF exhibited a higher rRNA synthesis rate when compared to young cells at similar growth rates resulting in the generally observed higher rRNA content (and cell size) of senescent cells. We conclude that cellular senescence of REF is not accompanied by rRNA expression deficiencies. PMID- 9402477 TI - Frailty of two cell cycle checkpoints which prevent entry into mitosis and progression through early mitotic stages in higher plant cells. AB - Allium cepa L. root meristems were given two short caffeine treatments spaced by 15 hours, the time which roughly corresponds to the duration of one cell cycle. In this way two subsequent cytokineses were prevented, and multinucleate cells with their in complement distributed into two, three or four nuclei were formed. Though all nuclei started to replicate synchronously in these cells, some of them (fast nuclei) completed their replication earlier than others (slow nuclei). The present report shows that two successive checkpoints operate before prometaphase in these cells. The first one prevents the entry of the fast nuclei into prophase until the slow ones have completed their replication. The second checkpoint ensures the synchronous entry into prometaphase after all nuclei have reached and finished prophase. By treating the multinucleate cells with an inhibitor of DNA synthesis at that time when fast but not slow nuclei had finished their replication, it was observed that both checkpoint mechanisms became leaky with time. Under these conditions the fast nuclei entered prophase in the presence of nuclei which were prevented from finishing the replication of their DNA. Subsequently, even prometaphase was triggered after a prolonged prophase. Finally, as expected from the presence of mitotic stages in these cells, nuclei with incompletely replicated DNA endured premature chromosome condensation. The prematurely condensed chromosomes either remained in a prometaphase-like stage until reconstitution nuclei formed or they followed the progression of the fast nuclei into metaphase and anaphase leading to the appearance of acentric chromosomal segments which after reconstitution gave rise to aneuploid nuclei containing unstable and broken DNA. PMID- 9402478 TI - Influence of 12(S)-hydroxyeicosatetraenoic acid (12(S)-HETE) on the localization of cathepsin B and cathepsin L in human lung tumor cells. AB - Cathepsins B and L are catabolic lysosomal enzymes but are likely candidates for extracellular proteolysis in normal and malignant processes. The signal mediator 12(S)-HETE selectively triggers a shot-gun release of cathepsin B. We have therefore investigated the intracellular distribution of cathepsins in unstimulated and 12(S)-HETE-stimulated tumor cells. Cathepsins B and L have only limited colocalization, which is found in the regions of synthesis and sorting (endoplasmic reticulum, Golgi, trans Golgi network). Treatment by 12(S)-HETE scatters cathepsin B but not cathepsin L and proform of cathepsin B. Colocalization with both mannose 6-phosphate receptors is very limited for both cathepsins. But extensive colocalization of cathepsin B and the endosomal/lysosomal marker CD63 (LIMP-I) documents the main fraction of the enzyme in these compartments. The supposed non-lysosomal fraction of cathepsin B is very likely the secretable material which follows a regulated secretory pathway. Storage and regulated secretion in tumor cells support extracellular proteolysis as a means in invasion which may lead to metastasis. But the mechanisms by which cells might acquire and eventually apply this means is still unknown. PMID- 9402479 TI - Phenoloxidase-dependent cytotoxic mechanism in ascidian (Styela plicata) hemocytes active against erythrocytes and K562 tumor cells. AB - The cytotoxic activity against rabbit erythrocytes (RE) and human K562 tumor cells by Styela plicata hemocytes was significantly related to the phenoloxidase (PO) which converts phenols to quinone and initiates the melanogenic pathway. The effector hemocyte population, separated in a Percoll density gradient band, enriched in a granulocyte type named "morula cells", was examined with RE in a hemocyte cytotoxic assay and plaque forming cell assay. Inhibition experiments with the copper chelating agents 1-phenyl-2-thiourea and tropolone, the substrate analogue sodium benzoate and sodium ascorbate support the notion that hemocyte cytotoxic activity is a PO-dependent mechanism. Treatments of hemocytes with the antioxidant enzymes, superoxide dismutase and catalase rule out oxy radicals produced by the melanogenic process as responsible of erythrolysis. Such a result suggests that quinone compounds derived from the melanogenic pathway might be the cytotoxic molecules. The PO-dependent anti-RE activity was also shown in a plaque forming assay in which "morula cells", containing polyphenols and PO, were identified as cytotoxic. PMID- 9402480 TI - Muscle biopsy for malignant hyperthermia screening in children. AB - Malignant Hyperthermia (MH) remains a life-threatening event in anaesthetic practice. In paediatric patients, triggering agents such as volatile anaesthetics and other succinylcholine are widely used. For children with a positive family history or previous clinical signs of MH, muscle biopsy for the halothane and caffeine in vitro muscle contracture tests is the only reliable procedure for diagnosis of MH susceptibility. Here we investigated outcome and compliance of patients and parents involved in the test. Twenty-four children between 6 and 14 yrs of age were admitted to hospital for biopsy. Muscle withdrawal was performed at the upper leg from lateral vastus muscle using regional or trigger-free general anaesthesia. Outcome and compliance were controlled by a telephone interview or direct physical re-evaluation. Seventeen patients out of 24 were diagnosed as clinically MH-susceptible according to the protocol of the European MH Group. Seven children were excluded as MH-negative by the test. Twenty-one children were evaluated postoperatively. Minor side effects of wound healing occurred, but none of the patients showed any abnormalities of muscle contracture or movement performance. Considering the high risk of fatal complications in the presence of MH-susceptibility, muscle biopsy of the upper leg for in-vitro diagnosis is a justified procedure that is acceptable to children and their parents. PMID- 9402481 TI - Characteristics of multisystem organ failure in neonates. AB - The records of 45 neonatal deaths in a four year period were reviewed retrospectively. Sixteen patients (35.5%) developed multisystem organ failure (MSOF). The criteria for pulmonary, hepatic, renal, hematologic, cardiac and microvascular failures were established. The onset of the first organ involvement was calculated in days prior to death. The earliest organ involved was kidney (14.2 +/- 15.1) followed by microvascular (9.4 +/- 7.6), hematologic (84. +/- 10.1), liver (6.8 +/- 6.7), lung (6.3 +/- 6.6) and cardiac (6.0 +/- 8.7) failure. Blood culture analyses revealed 5 patients with culture-positive sepsis. Yeast was the leading septic agent (n = 3) followed by Klebsiella pneumoniae (n = 2), Pseudomonas sp. (n = 1) and E. coli (n = 1). The first organ involvement was noted at 17.6 +/- 23.2 days. We concluded that the sequence of neonatal MSOF is different from that of adults, yet the inciting events are not clear-cut. Lung, which is the first organ involved in adults, seems to be a lately involved organ in neonates. PMID- 9402482 TI - Alpha-fetoprotein (AFP) levels in normal children. AB - Alpha-fetoprotein (AFP) is an important tumor marker for yolk sac tumor and hepatoblastoma in childhood. We have been using the graph of the normal range of serum AFP made by Tsuchida et al, when we evaluate the serum AFP levels in early infancy. We measured the serum AFP levels by an immunoradiometric assay in 163 normal infants under 2 years of age, in order to make a more precise graph. Our normal range was a little wider than that of Tsuchida et al. According to our graph, false-positive cases would be fewer. Referring to the half-lives of serum AFP levels in normal infancy is also useful, when it is difficult to evaluate the AFP level. PMID- 9402483 TI - The effect of hepatic vascular exclusion on hepatic blood flow and oxygen supply- uptake ratio in the pig. AB - The hemodynamic disturbances produced by total hepatic vascular exclusion (THVE) for 40 minutes were studied in 7 pigs (19-22 kg). THVE was produced by clamping the hepatic pedicle and inferior vena cava, above and below the liver, for a 40 minutes period, followed by unclamping. Compared to baseline values, 30 minutes after onset of THVE, there was a decrease in cardiac output (3.86 +/- 0.55 vs 1.23 +/- 0.23 L x min-1), systemic arterial pressure (97.54 +/- 13.58 vs 43.43 +/ 11.38 mm Hg), and pulmonary artery pressure (16.57 +/- 6.38 vs 12.57 +/- 3.58) and an increase in systemic and pulmonary vascular resistance (1772 +/- 198 vs 2351 +/- 462, and 182 +/- 66 vs 361 +/- 124 dyn x s x cm-5 respectively). As a result of diminished cardiac output, the systemic oxygen supply decreased (461 +/ 131 vs 101 +/- 46 ml x min-1), but the systemic oxygen extraction rate rose from 17.3% t0 31.2%. Thirty minutes after unclamping, the changes had reversed and all the parameters tended to normalize. Total hepatic blood flow 30 minutes after unclamping was higher than at baseline (5.08 +/- 1.2 vs 6.66 +/- 0.67 ml x min-1 x 100 g-1), because of the increase in portal blood flow (4.52 +/- 1.21 vs 6.07 +/- 0.70 ml x min-1 x 100 g-1). There were no significant differences in hepatic oxygen supply and uptake at baseline and after unclamping (152.6 +/- 23.0 vs 187.0 +/- 34.7 and 22.7 +/- 4.9 vs 28.7 +/- 8.4 ml O2 respectively). AST rose (29 +/- 7 vs 136 +/- 91 U/l), but there was no change in the remaining liver enzymes, glucose, creatinine and serum electrolytes, so we conclude that the hemodynamic disturbances produced by 40 minutes of THVE are manageable and spontaneously reversible. Liver metabolism was not greatly disturbed, so THVE was judged to be a viable technique to be added to the surgeon's range of options. PMID- 9402484 TI - Meconium ileus: a ten-year review of thirty-six patients. AB - Of 36 neonates with meconium ileus secondary to cystic fibrosis treated over a 10 year period, twenty-one (58%) had simple uncomplicated disease while fifteen (42%) had complications which included perforation (5), volvulus (6) and atresia (5). Gastrografin enema was employed in 20 infants with relief of obstruction in 8 (40%). Operative procedures consisted of resection and primary anastomosis in seventeen patients, stomas were fashioned in six, three had an enterotomy with irrigation only and two had Bishop-Koop enterostomy. Post-operative complications developed in 5 (18%) of these 28 patients. The overall survival rate was 97%. The one death occurred in an infant with short bowel syndrome, patent ductus arteriosus, hydrocephalus and pulmonary damage. There were eight additional patients who had meconium obstruction in the absence of cystic fibrosis. PMID- 9402485 TI - Effects of H2 receptor blocking agents on bacterial translocation in burn injury. AB - We experimentally studied the effects of H2 receptor blockers (ranitidine) on bacterial translocation (BT) in 42 male albino rats. Sham group (Group I, n = 12 rats) were exposed to 21 degrees C water while Burn group (Group II, n = 15 rats) and Ranitidine group (Group III, n = 15 rats) were exposed to 95 degrees C hot water for 10 seconds to produce a full thickness burn in 30% of total body surface area. 300 mg/kg ranitidine was administered to Group III starting immediately after the burn injury. Rats were sacrificed on the fifth postburn day. Sham group gained weight while groups II and III had significant weight loss. Gastric pH increased with the administration of ranitidine. Both gram negative and total number of bacteria were found to be reduced in cecal stool cultures in ranitidine group. Significant increase in BT was observed in Group III, and translocating bacteria were found to be different in burn and ranitidine groups with a final conclusion that administration of ranitidine changes intestinal ecological equilibrium and promotes BT. PMID- 9402486 TI - Compensatory renal growth post fetal nephrectomy in the rabbit. AB - Compensatory renal growth post-nephrectomy is well documented both clinically and experimentally. However, little is known about the capacity for compensatory growth in utero. We performed unilateral nephrectomy in fetal rabbits and studied the growth of the contralateral kidney. Thirty fetal rabbits underwent in utero uninephrectomies at day 25 of gestation. On gestational day 28, all the fetuses were delivered by cesarean section and the ratios kidney weight/body weight of the operated fetuses were compared to those of control littermates. The kidneys were then analysed by histology. A significant increase in renal weight was observed. The histological study of the remaining kidneys indicated a statistically significant increase of the glomerular area which confirmed the renal hypertrophy. This experiment demonstrates the capacity for the rabbit to develop in utero compensatory renal growth. PMID- 9402487 TI - Chromosomal aberrations in Wilms' tumour. AB - In 26 consecutive patients operated for Wilms' tumour samples from the tumour were genetically analyzed. Clonal acquired chromosome aberrations were found in 13 patients and a constitutional trisomy 18 as the sole change in 1. The chromosome number was altered in 13 patients. Numerical changes occurred in 16 patients and breakpoint of chromosome 1 in 6 patients. There was no structural alteration of chromosome 11. The observed cytogenetic heterogeneity illustrates the complexity of genetic changes involved in the genesis and progression of Wilms' tumour. To further elucidate the phenotypic impact of chromosomal aberrations the correlation to histology and the clinical course will be important. PMID- 9402488 TI - Microvascular autotransplantation of the testis: the "refluo" technique. AB - We think that the microvascular orchidopexy is the best technique for the treatment of the high intraabdominal testis. To prevent the problems related to the performing of the anastomosis between the spermatic and the inferior epigastric arteries we changed the traditional way for microvascular orchidopexy performing the venous anastomosis only, relying on the collateral deferential circle for the arterial supply; we called this technique the "refluo testicular autotransplantation". We supported our idea with an experimental investigation in rats and rabbits, with which we confirmed the efficacy of our proposal. This original technique, performed in 41 cases, allowed us to gain three important ends: a) reduction of the age at operation (under two years); b) reduction of the operating time (2 hours); c) improved success rate compared with the Fowler Stephens technique or staged orchidopexy. PMID- 9402489 TI - Surgical treatment of buried penis. AB - The buried penis is a rare congenital entity, whose treatment is surgical. There are few publications concerning this matter. The authors report on their experience in 10 cases (1990-1995). In this abnormality, the tip of the glans does not project from the pubic or scrotal skin. It is due to: 1) an excessive development of the penile fascia which retracts the penis; 2) insufficient attachment of the penile skin at the base of the penis; 3) often excessive prepubic fat worsens the appearance of the abnormality but does not by itself totally explain it; 4) a tight phimosis is often present. Surgical treatment is necessary because this aspect tends to persist even after puberty. One cannot indeed count on the development at the age of puberty, neither on the diminution of the fat, nor on the simple cure of the phimosis. One must above all ban circumcision which causes the risk of eliminating the skin necessary for reconstruction. The surgical procedure will comprise: 1) a longitudinal dorsal incision extended circumferentially; 2) resection of the thickened fascia penis; 3) anchoring of the deep face of the dermis to the proximal part of the fascia penis at the base of the penis. This surgical procedure has always brought a significant improvement to the appearance of the penis. PMID- 9402490 TI - Treatment of Madelung's deformity by lengthening and reaxation of the distal extremity of the radius by Ilizarov's technique. AB - Madelung's deformity was first described in 1878. It is characterised by a typical deformity of the carpus and not only causes pain but also impedes mobility and aesthetic appearance. Surgical correction can be effected during adolescence, the most frequently employed technique being conical osteotomy. We present a novel technique of lengthening and aligning the distal radial extremity using Ilizarov's technique. Five carpal joints were operated on in three 13-year old girls. An aesthetic effect was obvious in all the cases. Mobility improved by 30 degrees in the direction of the extension and pain always subsided directly after surgery. PMID- 9402491 TI - Ventriculoscopic implantation of ventricular shunts in children with hydrocephalus. AB - A modified technique of ventriculoscopy-aided implantation of ventricular shunts in children with hydrocephalus or intracranial cysts is presented. A 1 mm 0 degrees optic is introduced into the ventricle catheter and the shunt is positioned under video control. Finally, the optic is removed and the catheter left in place. Four endoscopy-aided implantations were performed with excellent visualization and without technical failure, the smallest patient weighted 680 grams. PMID- 9402492 TI - A case of non-functioning antiperistaltic retrosternal colic conduit replaced in situ and substituted with an isoperistaltic segment of ileum-caecum. AB - The authors report the case of a female patient, affected by long-gap oesophageal atresia, who, at 5 months old, was operated on for retrosternal substitution with a right-transverse antiperistaltic colic segment, in her local district hospital. Due to the anomalous position the neooesophagus never worked, and the baby was seriously dysphagic and failed to thrive. For this reason, when she was 11 months old, she was reoperated in our department. Through a medium sternolaparotomy the antiperistaltic colon was removed and replaced between the left and right colon; reconstruction was carried out with a retrosternal and isoperistaltic segment of ileo-caecum. The reoperation resolved her problems. This case is reported to confirm the author's opinion that all intestinal conduits must be positioned in the isoperistaltic direction. PMID- 9402493 TI - Bilateral adrenal neuroblastoma. AB - Neuroblastoma has been recognized as the most common solid tumor of infancy and childhood. The occurrence of bilateral adrenal neuroblastoma, however, is extremely rare and only a small number of cases have been previously reported. The authors herein report the clinical, histopathological and molecular biological features of two bilateral adrenal cases out of 125 neuroblastoma patients treated at Kyushu University Hospital over a 35-year period. The clinical and histopathological data of the two cases of bilateral adrenal neuroblastoma were reviewed. In Case 1, which had multiple liver metastases, a post-mortem examination revealed bilateral adrenal involvement. In Case 2, which had been detected by mass screening, CT showed masses in both adrenal glands. No big differences in size were recognized between the tumors in either of the cases. A histopathological examination revealed rosette fibrillary type neuroblastomas in both cases. The DNA of these tumor samples stored at -80 degrees C was extracted and the number of copies of the N-myc gene was determined by a Southern blot analysis. Fourteen copies of the gene were detected in Case 1, whereas neither of the tumors in Case 2 showed any amplification. The clinical outcome, histopathological findings and N-myc gene analysis of two cases might support the variety of biological features of this rare group of neuroblastoma. PMID- 9402494 TI - Synchronous occurrence of Wilms tumor and ganglioneuroblastoma in a child with neurofibromatosis. AB - A 2-year-old male with neurofibromatosis who had a Wilms tumor of the right kidney and an ipsilateral adrenal ganglioneuroblastoma is reported. Both tumors were completely removed and no recurrence occurred for 4 years after completion of the therapy. In a review of the literature, the prognosis of neurofibromatosis with these embryonal tumors is not satisfactory due to development of secondary tumors and disseminated metastases of the tumors. The synchronous occurrence of Wilms tumor and neuroblastoma in neurofibromatosis is extremely rare and this may be the first report in the world. PMID- 9402495 TI - Yolk sac tumor of the ovary in a conjoined twin. AB - A 5-year-old conjoined twin girl had a yolk sac tumor of the right ovary. She was admitted to our hospital because of abdominal distension. Comprehensive clinical and radiological investigations revealed a tumor of the right ovary. She underwent right salpingo-oophorectomy. The diagnosis was yolk sac tumor (stage I A), and postoperative chemotherapy (vincristine, actinomycine-D, cyclophosphamide) according to the UK-CCSG was provided. The postoperative course was uneventful with no evidence of recurrence 3 years after surgery. Her twin sister had no tumor. This is the first reported of yolk sac tumor of the ovary in a conjoined twin. PMID- 9402496 TI - Duplication of urethra--case report and review of literature. AB - Duplication of urethra is very rare. The duplicated urethra is always dorsal to the normal urethra which contains the sphincteric mechanism. A case is reported of urethral duplication where the duplicated urethra presented as a sinus on the dorsum of the penis. PMID- 9402497 TI - Carcinoma of the colon in childhood; report of 2 cases expressing p53 without K ras mutation. AB - We report on 2 children with colonic carcinoma and also review 62 cases of Japanese children with colonic carcinoma including ours. Although the dismal prognosis in colonic cancer in children is possibly due to the predominance of poorly differentiated carcinoma, there is no significant difference in the 5-year survival rates among well, moderately and poorly differentiated carcinomas in children. Positive staining with p53 in tumor cells was observed in each, but K ras mutations were not detected in any. Therefore, these carcinomas possibly developed from de-novo carcinoma. The development pathway of colonic carcinoma may relate to the prognosis in children, and be different from that in adults. PMID- 9402498 TI - Tooth mobility and periodontal disease. AB - Tooth mobility (TM) is an important feature of periodontal disease. This is evidenced by the large number of devices and methods of TM assessment that have been developed and tested. TM had been considered and investigated as an indirect measure of the functional condition of the periodontium as well as possible aggravating co-factor for periodontal disease. PMID- 9402499 TI - A study to investigate the effect of a propolis-containing mouthrinse on the inhibition of de novo plaque formation. AB - The results of a study to investigate the effectiveness of a propolis-containing mouthrinse in the inhibition of de novo plaque formation are presented. Subjects used a propolis-containing rinse, a negative control and a positive control in a double-blind, parallel, de novo plaque formation study design. The chlorhexidine mouthrinse was significantly better than the others in plaque inhibition. The propolis-containing rinse was marginally better than the negative control, but this difference was not significant. PMID- 9402500 TI - Substance P and neurokinin A in gingival crevicular fluid in periodontal health and disease. AB - The aims of the present study were to investigate whether the tachykinins substance P and neurokinin A were present in gingival crevicular fluid in both periodontal health and disease and to study the relationship with periodontal inflammation. Gingival crevicular fluid (GCF) was collected from a healthy, a gingivitis and a periodontitis site in 20 subjects with periodontitis and from a healthy site in 20 subjects without periodontitis. The volume of GCF was measured and each sample subsequently analysed for substance P and neurokinin A by radioimmunoassay. There were significantly increased levels of substance P-like immunoreactivity (SP-LI) and neurokinin A-like immunoreactivity (NKA-LI) in gingivitis and periodontitis sites compared with healthy sites. Both tachykinins were significantly elevated in periodontitis affected subjects, with significantly more tachykinin-like immunoreactivity at healthy sites in periodontitis affected compared with periodontally-healthy subjects. Despite the considerable individual variation in the levels of SP-LI and NKA-LI, both tachykinins were present at levels at which they could have biological activity. It is concluded that substance P and neurokinin A may have a role in the pathogenesis of periodontal disease and that further investigations could prove useful in clarifying the mechanisms through which neuropeptides could modulate periodontal health and disease. PMID- 9402501 TI - Amoxicillin and clavulanic acid concentrations in gingival crevicular fluid. AB - The beta-lactams are bactericidal antibiotics, but some of them may be inactivated by bacterial beta-lactamases which destroy the beta-lactam ring. The inactivation of amoxicillin by beta-lactamases of gram negative anaerobic bacteria can be circumvented by the addition of clavulanic acid, a beta lactamases inhibitor. Thus, most of these bacteria are susceptible to this combination. The aim of this study was to investigate the concentrations of amoxicillin and clavulanic acid in gingival crevicular fluid (GCF). These concentrations were measured in 20 patients with rapidly progressive periodontitis 1 h after a dose of 500 mg (1 tablet Augmentin) on day 0 and 1 h after the 10th intake on day 3. For the sampling of GCF, Periopapers were introduced in 16 gingival sites per subject and time. The GCF volumes collected were estimated using the Periotron 6000. A high performance liquid chromatography method has been developed for the determination of amoxicillin and clavulanic acid in microsamples (1 to 10 microliters) of GCF. The concentrations of amoxicillin and clavulanic acid were respectively, 14.05 micrograms ml-1 and 0.40 microgram ml-1 at day 0, 13.93 micrograms ml-1 and 0.37 microgram ml-1 at day 3. Effective levels of amoxicillin and clavulanic acid, well above the minimal inhibitory concentrations of some susceptible periodontal anaerobes (P. intermedia) involved in destructive periodontal diseases, are achieved following the multiple administration of amoxicillin combined with clavulanic acid. PMID- 9402502 TI - Guidelines for the design and conduct of clinical trials on dentine hypersensitivity. AB - Clinical trials on dentine hypersensitivity have been numerous and protocols varied. To date there is little consensus as to the conduct of studies on this poorly-understood yet common and painful dental condition. A committee of interested persons from academia and industry was convened to discuss the subject of clinical trials on dentine hypersensitivity and a consensus report is presented. A double-blind randomized parallel groups design is recommended, although cross-over designs may be used for the preliminary screening of agents. Subjects may have multiple sites scored. Sample size will be determined by estimating the variability in the study population, the effect to be detected and the power of the statistical test to be used. Subject selection is based on a clinical diagnosis of dentine hypersensitivity, excluding those with conflicting characteristics such as currently-active medical or dental therapy. The vestibular surfaces of incisors, cuspids and bicuspids are preferred as sites to be tested. A range of sensitivity levels should be included. Tactile, cold and evaporative air stimuli should be applied. Negative and benchmark controls should be incorporated. Most trials should last 8 weeks. Sensitivity may be assessed either in terms of the stimulus intensity required to evoke pain or the subjective evaluation of pain produced by a stimulus using a visual analog or other appropriate scale. The subject's overall assessment may be determined by questionnaire. Outcomes should be expressed in terms of clinically significant changes in symptoms. Follow-up evaluation is required to determine the persistence of changes. At least 2 independent trials should be conducted before a product receives approval. PMID- 9402503 TI - CD11b mRNA expression in neutrophils isolated from peripheral blood and gingival crevicular fluid. AB - Adhesion molecule CD11b/CD18 expressed by neutrophils (PMNs) participates in cell migration and phagocytosis of C3bi derivatized bacteria. It is this phagocytic function that eliminates some of the known periodontal pathogens in periodontal pockets. In patients with advanced periodontitis, homotypic aggregation of crevicular fluid PMNs (CF-PMNs) may occur due to overexpression of CD11b/CD18 and this may lead to ineffective elimination of periodontal pathogens. We have previously shown that CF-PMNs isolated from the periodontal pockets overexpress CD11b compared to PB-PMNs. This study tested the hypotheses that (1) overexpression of surface CD11b correlates with expression of CD11b mRNA in CF PMNs isolated from advanced periodontitis subjects, and (2) the intrinsic capacity of CD11b mRNA upregulation by PB-PMNs from periodontitis patients differs from that of control subjects. CF-PMNs and peripheral blood PMNs (PB PMNs) were isolated from 13 subjects with healthy gingiva (control group) and 13 subjects with advanced periodontitis (patient group). The surface expression of CD11b was determined by flow cytometry and CD11b mRNA was determined by extraction of mRNA and reverse transcription to cDNA followed by DNA amplification using primers to detect a segment of the cDNA which encodes CD11b. The results of this study confirm that the surface expression of CD11b on CF-PMNs is significantly higher in periodontitis subjects vs control subjects (p = 0.03), whereas surface CD11b expression on PB-PMNs does not differ significantly between groups (p = 0.06). The level of surface CD11b expression on CF-PMNs did not correlate with the amount of mRNA present in CF-PMNs in either group (p = 0.056, 0.07 for control and periodontitis patients, respectively). Most (9 of 13) individuals in the patient group expressed CD11b mRNA whereas very few control subjects (2 of 11) had CD11b mRNA in their CF-PMNs. This difference between groups was statistically significant (p = 0.004). The capacity to upregulate CD11b mRNA upon stimulation with fMLP and/or GM-CSF was highly variable and there was no statistical difference between the 2 groups. PMID- 9402504 TI - Reduced serum IgG reactivities with bacteria from dental plaque in HIV-infected persons with periodontitis. AB - Serum samples were obtained from 44 HIV-seropositive (HIV+) and 37 HIV seronegative (HIV-) persons that were grouped according to periodontal status. Serum IgG and IgA reactivities towards Streptococcus mutans, Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis. Prevotella intermedia, Prevotella nigrescens and Fusobacterium nucleatum were measured by means of ELISA. HIV+ persons with chronic marginal periodontitis showed significantly lower IgG reactivities to the periodontal pathogens A. actinomycetemcomitans, P. gingivalis, P. intermedia and F. nucleatum as compared with their HIV- counterparts (p < 0.05). Specific serum IgA reactivities were similar in the two periodontitis groups, except for P. nigrescens where the HIV+ group with chronic marginal periodontitis had lower values than their systemically healthy counterparts (p < 0.05). The results indicate that HIV infection affects the humoral serum immune responses against bacteria in dental plaque; the depressed antibody responses may contribute to the increased susceptibility for periodontal infections in HIV-infected patients. PMID- 9402505 TI - Comparison of the subgingival microbiota of periodontally healthy and diseased adults in northern Cameroon. AB - Our study is the 1st report on subgingival microbiota in adult Cameronians. The aim was to investigate, using the checkerboard DNA-DNA hybridization technique, the prevalence of 18 oral species in subgingival plaque samples obtained from sex and age-matched Cameronian adults with and without periodontal destruction. We also compared cultivation and the Affirm DP test with the checkerboard technique in their capability to detect some selected species among the 18. 21 adult periodontitis patients and 21 periodontally healthy subjects were examined and the results were compared statistically. Each periodontitis patient had at least 4 pockets of > or = 6 mm depth, while the healthy subjects had no sites more than 3 mm deep. Results of the checkerboard analysis showed that significantly (p < 0.05) more periodontitis patients tested positive for most of the 18 bacterial species. The Gram-positive species Actinomyces naeslundii, Streptococcus mitis and Streptococcus sanguis, known as microbiota of healthy sites, were detected significantly more frequently in the healthy group. Cultivation demonstrated P. gingivalis, B. forsythus, A. actinomycetemcomitans, P. intermedia and F. nucleatum in significantly lower %s of patients as compared to the checkerboard technique. Furthermore, the Affirm DP test detected P. gingivalis and B. forsythus in significantly fewer patients than did the checkerboard technique. A. actinomycetemcomitans was detected in 52.3% of the patients by the latter technique while the Affirm DP test failed to detect the bacterium in any of the samples. Overall, the results of the present study confirm the importance of the screening method and indicate that the prevalences of the investigated putative periodontal pathogens and beneficial species in the healthy and diseased adult Cameronians were similar to those reported for adults in the West and in some developing countries. PMID- 9402506 TI - Loss of deciduous teeth and germs of permanent incisors in a 4-year-old child. An atypic prepubertal periodontitis? A clinical, microbiological, immunological and ultrastructural study. AB - A 4-year-old child was referred, in April 1988, to Rennes Dental School (France) for deciduous tooth mobility with premature loss of 4 deciduous teeth and germs of 2 permanent incisors. Microbiological examinations by culture revealed the presence of the periodontal pathogen Actinobacillus actinomycetemcomitans. Immunofluorescence of plaque samples revealed the presence of Porphyromonas gingivalis that had not been isolated by culture. Neutrophil functions were within normal ranges. Transmission electron microscopy of gingiva showed a disorganised epithelium. The connective tissue was infiltrated by inflammatory cells. The basement membranes were normal, but the connective tissue-epithelium interface was mainly composed of short rete pegs. Scanning electron microscopy of extracted deciduous teeth revealed lack of cementum, lacunae in the cementum and lack of fibrillar insertion on the middle part of the root. Skin lesions, mainly situated on face, were observed. Treatment was by extraction of mobile deciduous teeth combined with 3-week courses of metronidazole. Clinical and microbiological follow-up was continued over a 7-year period. No periodontal lesions have been detected since eruption of the permanent teeth. The present subgingival and lingual microflora (December 1995) is composed of bacteria associated with periodontal health. However, the future appearance of a hitherto undetected systemic disease is still possible. PMID- 9402507 TI - A semiquantitative approach to the evaluation of acute cardiac allograft rejection at endomyocardial biopsy. AB - BACKGROUND: Histopathologic criteria for grading of acute cardiac allograft rejection are focused on the most severe lesion that is recognized among the myocardial fragments provided by each endomyocardial biopsy specimen. Considering the distribution of rejection lesions among all the fragments improved the accuracy in characterizing the severity of rejection in pathologic studies. This study was undertaken to verify the usefulness of a semiquantitative evaluation of endomyocardial biopsy specimens, consisting of the calculation of the proportion of fragments showing rejection in the clinical setting. METHODS: Of the 2386 biopsy specimens obtained during the first posttransplantation year in 168 consecutive cardiac allograft recipients, 290 biopsy specimens constituted by > or = 3 adequate fragments and showing rejection not followed by treatment (n = 159) or being the first biopsy specimen prompting treatment with augmented immunosuppression for that rejection episode (n = 131) were selected. These biopsy specimens (index biopsy specimens) were grouped according to whether rejection was present in < or = 33%, > 33% to < or = 67%, and > 67% of the fragments. The rejection grade (according to the standardized grading system) and the proportion of fragments positive for rejection were correlated with the occurrence of clinical symptoms and signs of rejection at index biopsy and with the results of the next biopsy. RESULTS: Rejections graded > or = 3A were more frequently symptomatic (36% vs 9% for those graded < 3, p < 0.0001), as were those involving increasing proportions of fragments (< or = 33%: 5 of 124, 4%; > 33 to < or = 67%: 13 of 99, 13%; > 67%: 19 of 67, 28% [p < 0.0001]). Spontaneous resolution after untreated biopsies was more frequent in focal (grade 1A and 2) than in diffuse mild (1B) rejections (68% vs 38% [p < 0.04]), whereas progression to grade 3A or greater was less frequent (4% vs 27% [p < 0.01]). Increasing proportions of positive fragments were associated with lower frequencies of spontaneous resolution (p < 0.05) and higher frequencies of worsening (9%, 22%, 43% [p < 0.009]) or progression to grade 3A or greater (2%, 6%, 28% [p < 0.005]). Complete resolution after treatment was less frequent for increasing proportions of positive fragments at index biopsy (80%, 66%, 49% [p < 0.05]). CONCLUSIONS: Diffuse versus focal rejection pattern and the proportion of positive fragments seem to be clinically relevant in terms of occurrence of symptoms, spontaneous evolution, and response to treatment. PMID- 9402508 TI - Efficacy of combining donor-specific presensitization with a simultaneous single injection of tacrolimus on pulmonary allografts. AB - BACKGROUND: We investigated the effects of combining donor-specific presensitization with a simultaneous injection of tacrolimus by use of a fully allogeneic rat lung allograft model. METHODS: Lungs from Brown Norway donor rats were orthotopically transplanted into Lewis recipient rats. Seven days before transplantation, allograft recipients received a transfusion of donor splenocytes (1 x 10(8) cells, intravenously), tacrolimus (3 or 1.5 mg/kg, intramuscularly), or a combination. No posttransplantation immunosuppression was given. In the transplantation study, graft survival was evaluated, and histologic characteristics of acute rejection were graded. In the in vitro studies, mixed lymphocyte reaction assays were used to investigate the effects of pretreatment on immune response. RESULTS: The graft survival evaluation disclosed that untreated rats rejected the allografts at 4.6 +/- 0.2 days. Donor splenocyte transfusion alone accelerated the graft rejection (3.3 +/- 0.2 days). Tacrolimus (3 mg/kg) alone moderately improved the graft survival (8.7 +/- 0.6 days). When donor splenocyte transfusion was combined with tacrolimus, graft survival was significantly increased to 29.6 +/- 12.0 days. In a mixed lymphocyte reaction assay, peripheral blood lymphocytes obtained from animals pretreated with donor splenocyte transfusion alone seemed to be hyperresponsive against the donor lymphocytes. In contrast, donor splenocyte transfusion with tacrolimus significantly suppressed the proliferative response against the donor lymphocytes but not against third-party lymphocytes obtained from naive Wistar King A rats. CONCLUSION: These data demonstrate that donor-specific presensitization with a simultaneous single injection of tacrolimus prevented both sensitization and graft rejection and induced donor-specific unresponsiveness. PMID- 9402509 TI - Long-term renal function in heart transplant recipients receiving cyclosporine therapy. AB - BACKGROUND: Immunosuppression with cyclosporine has improved allograft function and reduced both morbidity and mortality in organ transplantation. However, cyclosporine-induced nephrotoxicity still is a concern. The purpose of our study was to evaluate the effects of cyclosporine on renal function in orthotopic heart transplant recipients. METHODS: Thirty-nine patients who received transplants from 1985 to 1991 and had at least three yearly glomerular filtration rate measurements posttransplantation by 125I-iothalamate clearance method were included in the study. In addition, serum creatinine (before and after transplantation) and cyclosporine doses were analyzed. RESULTS: Maintenance immunosuppression at 1 year consisted of prednisone (0.1 mg/kg/day), azathioprine (2 mg/kg/day), and cyclosporine (12-hour trough level 100 to 150 ng/ml by fluorescence polarization immunoassay). The mean serum creatinine at 1 year was significantly higher than the mean pretransplantation serum creatinine (1.51 +/- 0.32 versus 1.28 +/- 0.38, p < 0.05) and stabilized after the first year. The mean glomerular filtration rate by 125I-iothalamate clearance method was 70.6 +/- 20.3 ml/min/1.73 m2 (range 32 to 105) at 1 year and remained relatively stable during the follow-up period of up to 7 years. Creatinine clearance calculated by the Cockcroft and Gault formula overestimated the true glomerular filtration rate after the third year. The mean cyclosporine dosage was significantly lower after the first-year dose of 3.9 +/- 1.8 mg/kg/day (p < 0.05). Three patients in 39 started hemodialysis at 5, 7, and 10 years after transplantation. CONCLUSION: Our data indicate that the adequacy of renal function is preserved with long-term cyclosporine therapy in heart transplant recipients. PMID- 9402510 TI - Causes of late failure after heart transplantation: a ten-year survey. AB - BACKGROUND: Little is known about the causes of death of heart transplant recipients who survive long-term. METHODS: The pathologic and clinical records of 97 patients who underwent heart transplantation in Italy from 1985 to 1995 and died (85 of 97) or underwent retransplantation (12 of 97) at least 2 years after transplantation were surveyed. Graft failures were classified as late (occurring between 2 and 5 years after transplantation) and belated (more than 5 years). RESULTS: Graft vasculopathy was the single most common cause of death (40.0%) and the only cause of late retransplantation. Tumors ranked second (23.5% of deaths), but the expected non-Hodgkin's lymphomas and Kaposi's sarcoma were accompanied by a high number of lung cancers (especially metastasizing adenocarcinomas). They were followed by the emergence or recurrence of pretransplantation diseases (9.4%), fatal infections (exclusively bacterial) (4.7%), the development of transmissible diseases (viral hepatitis and acquired immunodeficiency syndrome, 4.7%), and late acute rejection (2.3%). The distribution of failures differed in the late and belated periods: death and organ loss proportions for graft vasculopathy, respectively, fell and rose from the late to the belated period; some types of malignancy and fatal acute rejection were never observed in the belated period, whereas the emergence of pretransplantation diseases prevailed in the belated period. Graft vasculopathy was more frequent and tumors were less frequent among patients undergoing transplantation for ischemic heart disease. CONCLUSIONS: The reasons why heart transplant recipients die or undergo retransplantation, respectively, in the late and belated periods slightly differ from one another and are widely different than in short-term survivors. PMID- 9402511 TI - Overcoming right ventricular failure with left ventricular assist devices. AB - BACKGROUND: Right ventricular failure can lead to circulatory collapse while on left ventricular assist device support. By shunting blood from the femoral vein to the left ventricular assist device, cardiac output can be increased, but arterial oxygen saturation will decrease. METHODS: To determine the effects on O2 delivery, a model was developed on the basis of O2 uptake in the lungs and whole body O2 consumption. An equation was derived that related cardiac output, pulmonary venous O2 saturation, O2 consumption, and the ratio of shunt-to systemic blood flow to systemic O2 delivery. RESULTS: When total cardiac output increases, the shunt will increase systemic O2 delivery while decreasing arterial O2 saturation and leaving systemic venous O2 saturation unaltered. When total output does not increase, the shunt will decrease systemic O2 delivery, arterial O2 saturation, and systemic venous O2 saturation. CONCLUSIONS: The analysis suggests that measuring systemic venous oxygen saturation may be a useful way to monitor patient safety. A decrease in systemic venous O2 saturation when creating the shunt implies an inadequate increase in cardiac output. PMID- 9402512 TI - Cost-utility of lung transplantation: a pilot study. AB - BACKGROUND: The purpose of this study was to conduct a pilot investigation of the cost-utility of lung transplantation. With this study we provide a threshold analysis to estimate the survival gains that must be achieved for lung transplantation to be considered a beneficial use of society's resources. METHODS: A cross-sectional cohort design was used. All patients having undergone lung transplantation at the University of Pittsburgh Medical Center between March 1 and August 31, 1994, were identified via roster of transplant recipients (n = 20). Surviving patients were interviewed, by telephone, at their 1-year anniversary date. Utility was assessed by use of the quality of well-being scale. Direct cost of care was estimated from adjusted charges for the surgical admission, plus physician fees per the Medicare Physician Fee Schedule. RESULTS: The mean quality of well-being score for this group was 0.54 +/- 0.198 SD (median = 0.599, range 0 to 0.728). Summing the physician cost and the adjusted charges for the inpatient operative admission, the average cost of lung transplantation was $153,921 +/- $133,981 SD (median $94,324, range $63,405 to $598,482). At a cost of $94,324 and a utility of 0.599, the survival gain from surgery must be 2.7 years for the cost of the procedure to be justified from a societal perspective. CONCLUSIONS: Because of the many limitations in this pilot study, no firm policy implication may be drawn from these data. Directions for future research are discussed. PMID- 9402513 TI - Increased concentration of soluble human leukocyte antigen class I levels in the bronchoalveolar lavage of human pulmonary allografts. AB - BACKGROUND: Human leukocyte antigen (HLA) class I antigens, are glycoproteins expressed on the cell surface and are also secreted (sHLA) into the surrounding fluids. This study investigates whether pulmonary allograft rejection is associated with an increased amount of sHLA class I in the bronchoalveolar lavage (BAL) fluid. METHODS: Enzyme-linked immunosorbant assays were used to measure sHLA class I levels in BAL samples from 66 lung transplant recipients. RESULTS: Analysis of pulmonary allograft recipients revealed that the mean concentration of sHLA class I was significantly higher in samples from recipients with acute rejection than in recipients with no evidence of rejection. Seventy-two percent of the patients with rejection had sHLA levels above the normal range for patients with infection or rejection. Conversely, sHLA levels were within normal limits in 94% of the control population. Time kinetic analysis revealed that in subjects with rejection, sHLA class I levels peaked in the first 2 weeks after transplantation and decreased thereafter. Increased levels of sHLA were found in patients with acute rejection but not in those with chronic rejection. Fifty-nine percent (n = 10) of the patients with infection had sHLA levels within the normal (no infection or rejection) range. The remaining 41% (n = 7) with infection had sHLA above the normal range. Fifty-seven percent (n = 4) of the latter group had cytomegalovirus infection. These results were confirmed in a prospective study carried out by analyzing paired samples obtained during and after rejection. Eight of the nine patients had high BAL sHLA class I during acute rejection, and low BAL sHLA class I levels after resolution of the acute rejection. CONCLUSION: Our data demonstrate that measurement of sHLA class I levels in BAL samples is a sensitive indicator of acute rejection. PMID- 9402514 TI - Inhibition of accelerated coronary atherosclerosis with short-term blockade of intercellular adhesion molecule-1 and lymphocyte function-associated antigen-1 in a heterotopic murine model of heart transplantation. AB - BACKGROUND: Graft arteriopathy limits the long-term survival of allograft recipients. Cardiac allografts in mice develop graft coronary arteriopathy similar to that observed in clinical chronic rejection in human beings. We found that antiintercellular adhesion molecule-1 (ICAM-1) and antilymphocyte function associated antigen-1 (LFA-1) monoclonal antibodies (mAbs) induce immunologic tolerance to mice with cardiac allografts. METHODS: To evaluate the effects of short-term administration of anti-ICAM-1 plus anti-LFA-1 mAbs in preventing graft arteriopathy, we treated C3H/He mice that received cardiac allografts from BALB/c mice with anti-ICAM-1 plus anti-LFA-1 mAbs for the first 5 days after transplantation. For control studies, FK506 was administered daily to other allograft recipients. Allografts were harvested on day 60. Immunohistochemical analysis was used to detect the expression of ICAM-1 and vascular cell adhesion molecule (VCAM)-1, and in situ reverse transcriptase polymerase chain reaction was performed to detect platelet-derived growth factor (PDGF)-B mRNA expression in the graft arteries. RESULTS: Allografts from mice that received FK506 treatment daily showed significant neointimal thickening with increased expression of ICAM-1, VCAM-1, and PDGF-B mRNA, whereas there was almost no intimal thickening and ICAM-1, VCAM-1, and PDGF-B mRNA expression in the mice that received anti-ICAM-1 plus anti-LFA-1 mAbs. CONCLUSION: Short-term blockade of ICAM-1 and LFA-1 adhesion not only induces immunologic tolerance to cardiac allografts but also prevents graft arteriopathy. PMID- 9402515 TI - Assessment of tolerance induction to cardiac allograft by anti-ICAM-1 and anti LFA-1 monoclonal antibodies. AB - BACKGROUND: Transplantation tolerance is induced by selective manipulation of intercellular adhesion molecule-1 (ICAM-1) and leukocyte function-associated antigen-1 (LFA-1) molecules in the adult mouse. However, the mechanism of this tolerance induction has not been elucidated. METHODS AND RESULTS: C3H/He mice were heterotopically transplanted with BALB/c hearts; recipients were injected with anti-ICAM-1 and anti-LFA-1 monoclonal antibodies (mAbs). Of 3-day administration of both mAbs at 50 micrograms/day, all recipients accepted cardiac allografts indefinitely (n = 20); this also occurred in 5 of 10 mice that were not treated until day +4 or day +5. Long-term cardiac allograft acceptance was also achieved in thymectomized recipients. This tolerance could not be abrogated by the injection of either naive recipient lymphocytes or sensitized lymphocytes taken from the recipient strain mice that rejected the same donor strain hearts. Mixed lymphocyte culture showed that splenocytes from allograft recipient mice treated with the mAbs showed normal allogeneic response without donor alloantigen specificity. CONCLUSIONS: These results demonstrate the unique character of the peripheral tolerance induced by anti-ICAM-1 and anti-LFA-1 mAbs. Further studies are required to elucidate detailed mechanisms of transplantation tolerance development. PMID- 9402517 TI - Differential pulmonary flow after heart transplantation in patients with malposition of the great arteries. AB - In transposition of the great arteries the right lung is preferentially perfused, and this may be due to the orientation of the right pulmonary artery when the main pulmonary artery comes off in a posterior position. In this study the mean perfusion of the right lung was 61% in six patients who had "correction" of the great artery malposition at the time of heart transplantation. This suggests that anatomic correction of pulmonary artery orientation does not correct the preferential perfusion to the right lung. PMID- 9402516 TI - Clinical correlates of the myocardial force-frequency relationship in patients with end-stage heart failure. AB - BACKGROUND: This study tested the hypothesis that in patients with suspected heart failure, peak oxygen consumption was the best predictor of heart muscle failure. Failing human myocardium is characterized by an abnormal force-frequency relationship, which has been previously shown to be altered in parallel with the severity of heart failure. METHODS: We examined whether seven different functional parameters of isolated electrically driven ventricular trabeculae carneae obtained from 34 explanted hearts of patients undergoing heart transplantation for end-stage heart failure correlated with any of 47 separate pretransplantation clinical parameters. The functional muscle parameters were active force at 0.33 Hz, time to 80% relaxation (RT 80%) of twitch force at 0.33 Hz, optimal frequency (OF), active force at 1.0 Hz (AF1), diastolic force at 1.0 Hz (DF1), active force at 2.0 Hz (AF2), and diastolic force at 2.0 Hz (DF2). RESULTS: Before transplantation the mean left ventricular ejection fraction was 21% +/- 10%, and all patients were in New York Heart Association class III or IV. Mean peak whole body VO2 was 10.9 +/- 3.3 ml/min/kg and percent body mass/age/sex adjusted maximum VO2 oxygen consumption was 34.7% +/- 10.4%. Univariate analysis of VO2 yielded the following significant correlations: active force at 33 Hz, RT80%, OF, AF1, DF1, AF2, DF2; whereas univariate analysis of percent body mass/age/sex-adjusted VO2 yielded the following significant correlations: RT80%, OF, DF1, AF2, DF2. Multivariate analysis showed that OF and DF1 were independent predictors of peak VO2. CONCLUSION: In this study we show that peak oxygen uptake measured during cardiopulmonary exercise testing obtained before transplantation is correlated with the force-frequency behavior of isolated muscles at the time of transplantation. Peak VO2 seems to be a strong indicator of the severity of cardiac contractile dysfunction in patients with heart failure. PMID- 9402518 TI - Cytomegalovirus infection presenting as an apple-core lesion of the colon. AB - Cytomegalovirus infection is highly prevalent among heart transplant recipients. Symptomatic cytomegalovirus infection can occur in all parts of the gastrointestinal tract. Colonic lesions are usually manifest as hemorrhagic colitis. This is a case of cytomegalovirus colitis presenting as a colonic stricture mimicking a colonic carcinoma. The initial presentation was that of both cellular and humoral rejection with fever, abdominal pain, and microcytic anemia with heme-positive stools. An abdominal computed tomogram was pertinent for a suspicion of carcinoma in the midtransverse colon. After resolution of the rejection episode, colonoscopy was performed, the result of which was abnormal for a short, high-grade stricture in the midtransverse colon. The patient underwent a right hemicolectomy for the suspected tumor. The pathologic specimen showed cytomegalovirus inclusion bodies with acute suppurative ulceration. The early diagnosis and treatment of cytomegalovirus colitis may lead to avoidance of more serious complications such as stricture formation. PMID- 9402519 TI - Successful mechanical circulatory support for more than two years with a left ventricular assist device in a patient with dilated cardiomyopathy. AB - A 36-year-old man with dilated cardiomyopathy was supported with a left ventricular assist device for 795 days. During this support time gradual functional recovery was noted. The patient tolerated the device well, enjoyed a good quality of life, and experienced no technical device-related problems. When a suitable donor heart became available, the device was switched off, and native ventricular function was assessed. This was judged to be acceptable, and the left ventricular assist device was successfully explanted. Postexplantation function remained acceptable and improved over the following days. The patient was discharged on the twentieth postoperative day and remains well. PMID- 9402520 TI - Left ventricular assist devices: permanent implant versus bridge to transplantation--is either cost-effective? PMID- 9402521 TI - SIADH and acute lung rejection. PMID- 9402522 TI - Characterization of the phosphoribosylpyrophosphate synthetase gene from Candida albicans. AB - Phosphoribosylpyrophosphate (PRPP) synthetase catalyzes the transfer of phosphate from ATP to D-ribose-5-phosphate during the synthesis of purine and pyrimidine nucleotides and tryptophan and histidine biosynthesis in a variety of organisms. We cloned and sequenced the PRPP synthetase gene (PRS1) of Candida albicans because, in Saccharomyces cerevisiae, a deficiency in PRPP synthetase activity interacts with a mutation in ELM4-1 (elongated morphology) to cause constitutive pseudohyphal growth in nitrogen-rich media. In order to study the role of the C. albicans PRS1 in growth and morphogenesis, we used gene disruption to isolate PRS1 mutants; however, while heterozygous PRS1 clones were readily obtained, homozygous, null strains were not recovered indicating that PRS1 is probably essential for growth of the organism. Heterozygotes in PRS1 produced approximately 35% less PRPP synthetase (P = 0.0004) and exhibited a similar reduction in transcript levels. Confirmation of a heterozygous, single disruption in PRS1 was obtained by I-SceI digestion of chromosomal-sized DNA and Southern blot hybridizations. While no role in morphogenesis is elucidated by this work, the data strongly suggests that PRS1 is an essential gene in C. albicans and supports earlier results that indicated the presence of a single PRS gene in C. albicans. PMID- 9402523 TI - Study of Cryptococcus neoformans actin gene regulation with a beta-galactosidase actin fusion. AB - An expression plasmid carrying a heterologous gene fusion between the Cryptococcus neoformans actin promoter and the Escherichia coli reporter gene, LACZ, was constructed to study actin regulation in C. neoformans. Two randomly stable transformants, designated 20.6 and 20.9, were selected for further examination. Both ectopic and homologous recombination with vector insertion in tandem repeats occurred in these transformants. Transformant 20.9 carried more copies of ACTp::LACZ in its genome than 20.6 and this was reflected in expressing higher levels of beta-galactosidase activity. In vitro, these transformants showed higher levels of beta-galactosidase activity expressed when the transformants were propagated at higher temperatures (37 degrees C vs 30 degrees C). However, beta-galactosidase expression in the transformants was variable during logarithmic and stationary growth phases and this differential expression was temperature dependent. This report shows that the constitutive actin gene in C. neoformans is regulated by temperature and growth and this fact should be taken into consideration when actin expression is used as a standard to compare the expression of other regulated genes. Also, a more sensitive reporter construct will be needed for in vivo gene analysis of regulation. PMID- 9402524 TI - Coccidioidomycosis in Tulare County, California, 1991: reemergence of an endemic disease. AB - In 1991, 1208 cases of coccidioidomycosis were reported to the California Department of Health Services, compared with an annual average of 450 during 1986 90. We conducted a study in Tulare County to define the epidemiology of the disease and identify risk factors for severe disease, focusing on the epidemic period September 1991-December 1991. To identify cases, we used data from the Coccidioidomycosis Serology Laboratory at the University of California, Davis, other laboratories, and the Tulare County Health Department's coccidioidomycosis reporting system. We compared patients who were hospitalized with those who were not to determine risk factors for severe disease. We identified 128 cases of acute coccidioidomycosis diagnosed between 1 September and 31 December 1991 (attack rate 41/100,000); south central Tulare County had the highest attack rate. Thirty-five (27%) case-patients were hospitalized. Male sex (relative risk (RR) 2.5, 95% confidence interval (CI) 1.2-5.0), black people and Asian races (RR 4.8, 95% CI 2.4-9.6), and age > or = 20 years (RR 8.3, 95% CI 1.2-57.4) were univariately significant and remained independently associated with hospitalization in multivariate analysis. The 1991 Tulare County outbreak of coccidioidomycosis was part of a much larger outbreak that began in California during 1991 and continued through 1993. The outbreak was preceded by an unusually rainy spring. Although dust reduction measures during times of increased coccidioidomycosis incidence can help reduce exposure, definitive control awaits the development of a safe, effective vaccine. PMID- 9402526 TI - Ascosubramania gen. nov., and its Fonsecaea-like anamorph causing chromoblastomycosis in India. AB - This paper deals with the discovery of the teleomorph of a Fonsecaea-like pathogen causing chromoblastomycosis in India. A new genus, Ascosubramania, is proposed to accommodate it. PMID- 9402525 TI - Induction of interleukin-6 mRNA in rat alveolar macrophages by in vitro exposure to both Cryptococcus neoformans and anti-C. neoformans antiserum. AB - Lewis rat alveolar macrophages (AM) were harvested and exposed in vitro to Cryptococcus neoformans to investigate the induction of inflammatory cytokines. AM in tissue culture wells were incubated with viable yeast cells of C. neoformans or the capsular polysaccharide, glucuronoxylomannan (GXM), with or without rabbit anti-C. neoformans antiserum. At 3, 6, 12 and 24 h, AM were washed, lyzed and total RNA was isolated. Using reverse transcription-PCR, the transcripts of cytokine genes were semi-quantified by comparison with constitutive transcripts. Incubation of AM with lipopolysaccharide, as positive control, induced elevated levels of the three transcripts measured: interleukin (IL)-1 alpha, IL-6 and tumour necrosis factor (TNF)-alpha. Under the same conditions, no obvious changes were observed in the levels of transcription of these cytokines by AM after exposure to several strains of C. neoformans. However, AM that were incubated with both the yeast cells and rabbit polyclonal antisera to C. neoformans manifested significantly increased levels of mRNA for IL-6, but not IL-1 alpha or TNF-alpha. This increased level of IL-6 mRNA was detectable after incubation for 6 or 12 h. Levels of transcription in AM were unaffected by exposure to normal rabbit serum, specific antiserum alone. GXM at concentrations of 10, 100 or 500 micrograms ml-1, or GXM and antiserum. Adsorption of the antiserum with heat-killed yeast cells of C. neoformans diminished its ability to induce IL-6 mRNA in combination with fresh, viable yeast cells. The induction of IL-6 mRNA by yeast cells and antiserum does not require intact complement. In the absence of complement, the rabbit antiserum served as a potent opsonin and markedly increased phagocytosis of C. neoformans by AM. These results indicate that antibody-opsonized C. neoformans are readily phagocytosed by rat AM, and that antibody-mediated phagocytosis may differ from complement-mediated phagocytosis in the subsequent stimulation of IL-6. PMID- 9402527 TI - Expression of glycoprotein gp43 in stage-specific forms and during dimorphic differentiation of Paracoccidioides brasiliensis. AB - Expression of the 43 kDa glycoprotein (gp43) was analysed in several Paracoccidioides brasiliensis isolates. Using one- and two-dimensional analysis of crude cellular extracts, it was shown that protein expression in yeast and mycelium was dependent on the isolate analysed. In two strains, in both yeast and mycelium cells. gp43 was present, whereas expression was restricted to the yeast phase of two other strains. The clinical implications of this phase-specific gp43 expression are uncertain. PMID- 9402528 TI - Histopathological and electron microscopical studies on experimental Penicillium marneffei infection in mice. AB - Experimental Penicillium marneffei infection in mice was investigated histopathologically and by electron microscopy. Viable conidia (5 x 10(6) cells) of P. marneffei were inoculated into each mouse of group A (BALB/c mice) and group B (BALB/c-nu/nu-SIc mice) through the tail vein. All the mice were sacrificed at intervals and the livers were examined. In group A, the conidia were phagocytosed by Kupffer cells soon after inoculation, and proliferated by fission in the cytoplasm. Marked proliferation of yeast cells was observed 7 and 14 days after inoculation. With proliferation of the fungus, the number of lysosomes in Kupffer cells increased, and numerous granulomas were formed in the liver. These granulomas consisted mainly of macrophages with yeast cells, together with a few polymorphonuclear leukocytes, lymphocytes and giant cells. From 28 days on yeast cells were gradually cleared from the granulomas, and 56 days after inoculation almost all the granulomas disappeared. In group B, at an early stage of infection, similar pathological changes to those seen in mice of group A were observed. However, as the infection progressed, the number of granulomas continued to increase and yeast cells continued to proliferate although lymphocytes did not infiltrate these granulomas. With proliferation of yeast cells the liver tissue was replaced with both yeast cells engulfed by macrophages and extracellular yeasts, and dissemination occurred. PMID- 9402529 TI - Identification by polymerase chain reaction fingerprinting of Cryptococcus neoformans serotype AD. AB - Seventy-three Cryptococcus neoformans isolates and eight other yeast strains were studied. Fingerprints produced by priming with (GACA)4 differentiated C. neoformans from all other yeasts tested and identified the five C. neoformans serotypes. Four major bands of molecular size 800, 540, 475 and 410 bp were recognized for serotypes A, AD and D. Two of them were specific for serotype A and the other two for serotype D isolates. Serotype AD strains were identified by five different genotypic patterns in which at least one of the two bands specific for serotype A and D were present in different combinations. On repeated and simultaneously performed genotype and serotype testing of nine strains, the genotypic pattern did not change, whereas serotyping was unstable in three cases. PCR-fingerprinting using (GACA)4 as a primer proved more stable than serology in discriminating among C. neoformans serotypes A, D and AD and was able to distinguish among serotype AD strains. PMID- 9402530 TI - Fungal keratitis caused by Metarhizium anisopliae var. anisopliae. AB - Metarhizium anisopliae var. anisopliae (Metschnikov) Sorokin 1883 to our knowledge has never been reported as an agent of human or animal mycosis. This fungus has great importance as an agent of biological control of different pests and mosquito larvae in Colombia. It has been isolated as the aetiological agent of keratomycosis for the first time from the eye of a Colombian male. PMID- 9402531 TI - Maxillary sinusitis caused by Schizophyllum commune and experience with treatment. AB - A case of sinusitis caused by the basidiomycete Schizophyllum commune is reported in a 36-year-old female with a history of allergic rhinitis and dermatitis. The patient presented with sudden nasal obstruction, purulent nasal discharge, headache and general discomfort. Computer tomography revealed extensive opacity of the left maxillary sinus as well as erosion of the nasal wall and maxillary bone. Mycological examinations of nasal discharges and material aspirated during anthrostomy showed hyaline, septate hyphae with rare spicules. Primary isolation yielded a white, woolly mould which demonstrated clamp connections and basidiocarp primordia but these characteristics were lost in subculture. Identification was confirmed by vegetative compatibility studies. The patient was treated with itraconazole to avoid possible postsurgical dissemination. Three months after cessation of therapy, no recurrence of infection had occurred. PMID- 9402533 TI - In vitro susceptibility of Malassezia furfur to the essential oil of Melaleuca alternifolia. AB - The susceptibility of 64 Malassezia furfur isolates to Melaleuca alternifolia oil was determined. The minimum inhibitory concentration for 90% of isolates was 0.25% by agar dilution and 0.12% by broth dilution. These data indicate that tea tree oil may be useful in the treatment of skin conditions involving M. furfur. PMID- 9402532 TI - Detection of plasma (1 --> 3)-beta-D-glucan in patients with Fusarium, Trichosporon, Saccharomyces and Acremonium fungaemias. AB - (1 --> 3)-beta-D-glucan, a characteristic fungal molecule, is known to increase in blood in invasive candidiasis, aspergillosis and cryptococcosis. This report shows that the plasma glucan concentration was also elevated in four patients infected with Fusarium, Trichosporon beigelii, Saccharomyces cerevisiae and Acremonium. In three of the patients, the elevation preceded positive blood cultures by 5-17 days, and in one of them it even preceded the onset of fever by 6 days. In a fourth patient, the glucan level decreased with clinical improvement. Plasma (1 --> 3)-beta-D-glucan determination appears to be useful also for diagnosis and follow-up of these unusual deep mycoses. PMID- 9402534 TI - Asymptomatic sequelae to acute sarin poisoning in the central and autonomic nervous system 6 months after the Tokyo subway attack. AB - Six to eight months after the Tokyo subway attack in March 1995, the neurophysiological effects of acute sarin poisoning were investigated in 18 passengers exposed to sarin (sarin cases) in the subways to ascertain the focal or functional brain deficits induced by sarin. The event-related and visual evoked potentials (P300 and VEP), brainstem auditory evoked potential, and electrocardiographic R-R interval variability (CVRR), together with the score on the posttraumatic stress disorder (PTSD) checklist, were measured in the sarin cases and the same number of control subjects matched for sex and age. None of the sarin cases had any obvious clinical abnormalities at the time of testing. The P300 and VEP (P100) latencies in the sarin cases were significantly prolonged compared with the matched controls. In the sarin cases, the CVRR was significantly related to serum cholinesterase (ChE) levels determined immediately after exposure; the PTSD score was not significantly associated with any neurophysiological data despite the high PTSD score in the sarin cases. These findings suggest that asymptomatic sequelae to sarin exposure, rather than PTSD, persist in the higher and visual nervous systems beyond the turnover period of ChE; sarin may have neurotoxic actions in addition to the inhibitory action on brain ChE. PMID- 9402535 TI - Dissociated neglect for objective and subjective sizes. AB - We examined the effect of line length and viewing distance on the line bisection performance in near space in five patients with left unilateral spatial neglect following right parietal lesions. A line bisection task by fixation was devised to avoid the influence of manual responses. The rightward deviation measured in visual angle increased linearly as a function of the visual angle of lines 150 mm or more long. This linearity, however, did not hold for lines of 100 mm or less. The deviation measured in length was nearly constant for each of these short lengths, even when the visual angle was varied at different viewing distances. The patients therefore discriminated the objective lengths of the short lines. For small objects, neglect patients may distribute attention mainly on the coordinates scaled for objective size. PMID- 9402536 TI - Abnormal calcium metabolism in myotonic dystrophy as shown by the Ellsworth Howard test and its relation to CTG triplet repeat length. AB - Myotonic dystrophy (DM) is an autosomal dominant disorder characterized by peculiar clinical features. Its molecular basis is the unstable expansion of a CTG triplet repeat in the gene encoding myotonin protein kinase (Mt-PK), the nucleotide sequence of which has extensive homology to the cyclic AMP (cAMP) dependent protein kinase gene. Extensive efforts have been made to clarify the signal transduction pathway in which the responsible gene operates, but confirming evidence has yet to be obtained. Because some symptoms in DM are similar to those in hypoparathyroidism, we divided 24 DM patients into two groups on the basis of their serum calcium levels; Group 1, those with normocalcemia (11 patients), and group 2, those with hypocalcemia (13 patients). The highly sensitive parathyroid hormone (HS-PTH) plasma levels in group 1 were within normal limits, whereas those in group 2 were abnormally high. Laboratory findings for the group 2 patients resembled those for pseudohypoparathyroidism (PHP), whereas those for group 1 patients were normal. The Ellsworth-Howard (EH) test was used to determine which type of PHP the group 2 patients belonged to. Both the phosphaturic (delta P) and urinary cAMP (UcAMP) responses were estimated. The delta P responses in group 2 were significantly lower than those in group 1, but their UcAMP responses did not differ. This is evidence that group 2 patients had PHP type II, whereas group 1 patients were normal. We also investigated whether the disease severity differed between the groups. Cataracts, ectopic calcifications, and ossifications, which are associated with PHP, were more frequent in group 2. In addition, the mean IQ in that group was significantly lower. Clinically, the group 2 signs agreed well with those of PHP, whereas for group 1 there was only a slight similarity. These results are additional evidence that the patients in group 2 have abnormal calcium metabolism, the abnormality being in the postadenylate cyclase-cAMP pathway in the renal tubular cells. The degree of (CTG)n expansion, the so-called expanded DNA fragment (EF) size, was determined by standard Southern blot analysis. The allelic EF sizes in both groups were greater than in the healthy controls. Moreover, those in group 2 were significantly longer than those in group 1. We therefore investigated whether EF size is correlated with the serum calcium and plasma PTH levels, the delta P responses in the EH test, and IQ. All these items were significantly correlated with EF size. Our findings show that the expanded DNA fragment size in DM is correlated with the degree of abnormal calcium metabolism. PMID- 9402537 TI - Psychogenic pseudoptosis. AB - Three patients with psychogenic pseudoptosis of one eyelid are reported. All showed depression of the eyebrow on the affected side. The clinical course varied: in two patients the symptom resolved spontaneously after positive reassurance; in the third patient it remained unchanged for 2 years. PMID- 9402539 TI - Facial palsy in multiple sclerosis. AB - Facial palsy occurred in 21 (19.6%) of 107 Japanese patients with multiple sclerosis (MS) during a mean follow-up period of 4.3 years. We observed residual signs of facial palsy in five other patients in whom acute onset was confirmed from medical records. Facial palsy began on average 7.6 years after the onset of MS but in five patients (4.7%) was the first symptom of MS, preceding the next MS symptom by 0.5-3 years. Facial palsy was usually associated with other brainstem signs, while two patients showed only facial palsy 1 and 3 years after the onset of MS. Twenty-one (84.0%) of the 25 patients who underwent brain magnetic resonance imaging (MRI) showed brainstem lesions in the pontine tegmentum ipsilateral to the facial palsy. However, the two patients without other symptoms or signs had no apparent causal lesion on MRI, which suggests difficulty in differentiating idiopathic Bell's palsy from MS- associated facial palsy by MRI, although it has an excellent capacity to detect causal lesions of facial palsy associated with MS. PMID- 9402538 TI - Criteria for early detection of conduction block in multifocal motor neuropathy (MMN): a study based on control populations and follow-up of MMN patients. AB - Motor conduction block (MCB) has been used as the main diagnostic criterion in multifocal motor neuropathy (MMN). Nonetheless, no agreed definition of block currently exists; the proposed required percent decrement of proximal compound muscle action potential (CMAP) amplitude varies from > 20% to > 50%. The aim of this work was to evaluate, through a follow-up study of patients with MMN, the behaviour of MCB over time. The percent decrement and temporal dispersion of proximal CMAP have also been calculated in normal controls and in patients affected by amyotrophic lateral sclerosis (ALS). The results show that MCB in patients with MMN is a dynamic entity which greatly varies over time and that a > 50% CMAP amplitued reduction may well be preceded by a smaller decrement that is nonetheless indicative of focal myelin damage in the appropriate clinical context. This datum and the results obtained in the control group and in patients with ALS suggest that a reappraisal of the diagnostic criteria for MCB, in cases with clinical and electrophysiological data strongly indicative of MMN, should be considered. Since MMN is a treatable disorder, the use of the proposed less restrictive criteria for the identification of MCB could allow for a promp and more effective treatment. PMID- 9402540 TI - Level of transforming growth factor beta 1 is elevated in cerebrospinal fluid of children with acute bacterial meningitis. AB - We investigated the levels of transforming growth factor beta 1 (TGF-beta 1) in cerebrospinal fluid (CSF) in children with meningitis, with a view to prognostic relevance. CSF TGF-beta 1 levels on admission were measured by a sandwich enzyme immunoassay in children with bacterial meningitis (n = 16), aseptic meningitis (n = 12), and control subjects without evidence of central nervous system (CNS) infection (n = 16). Patients were followed up for a mean duration of 13 months, and neurodevelopmental sequelae was determined for those with bacterial meningitis. On admission, CSF TGF-beta 1 levels were significantly higher in children with bacterial meningitis (mean, standard error, 32.92, 2.36 pg/ml) as opposed to those with aseptic meningitis (25.26, 1.72 pg/ml) (P = 0.0155), or control subjects (20.53, 1.05 pg/ml) (P < 0.0001). The CSF TGF-beta 1 levels in children with aseptic meningitis were higher than those in the control group, but without significance (P = 0.02). No apparent correlation existed between CSF TGF beta 1 levels and CSF protein or cell counts in patients with bacterial meningitis. No significant difference in CSF TGF-beta 1 levels was found between patients with or without major sequelae following bacterial meningitis. PMID- 9402541 TI - Ocular palsies in the absence of other neurological or ocular symptoms: analysis of 105 cases. AB - We studied prospectively 105 unselected patients complaining of ptosis and/or diplopia due to extrinsic ophthalmic muscle palsies without other neurological signs. All patients underwent the same diagnostic protocol. The presenting symptoms were: ptosis, 35 patients (33%); diplopia, 27 patients (26%); ptosis and diplopia, 43 patients (41%). The oculomotor nerve was most frequently involved, followed by the abducens nerve. The final diagnoses were: ocular myasthenia, intracranial and/or orbital pathology, thyroid ophthalmopathy, diabetic ophthalmoplegia, mitochondrial myopathy, oculopharyngeal muscular dystrophy. In 26 patients (25%) the cause remained undetermined. Our study confirms the difficulty of establishing an aetiological diagnosis in patients with isolated ocular palsies. PMID- 9402542 TI - Difference in P300 latency in two types of leukoaraiosis. AB - P300 examination was performed in patients with periventricular leukoaraiosis and in patients with leukoaraiosis in the centrum semiovale. Ten patients with periventricular leukoaraiosis, ten patients with leukoaraiosis in centrum semiovale and ten age-matched controls without leukoaraiosis were studied. The P300 was measured with an evoked potential recorder using an oddball paradigm. The mini-mental state examination score was significantly lower and the P300 latency was significantly longer in the leukoaraiosis in the centrum semiovale group than in the periventricular leukoaraiosis group and the control group. Leukoaraiosis in centrum semiovale may be related to dementia and the prolongation of P300 latency. PMID- 9402544 TI - Evolution of cardiac abnormalities in Becker muscular dystrophy over a 13-year period. AB - We evaluated the course of cardiac involvement in 27 previously reported patients with Becker muscular dystrophy (BMD) originating from nine kindreds. Since almost all affected individuals of each kindred were included, intrafamilial variability could be studied. We also attempted to identify associations between cardiac involvement, functional ability and mutations at DNA level. The mean follow-up period was 12.5 years. The number of patients with electrocardiographic abnormalities progressed from 44% to 71%. Dilated cardiomyopathy (DCM) with or without congestive heart failure was now present in 33% as compared with 15% in the previous study. In addition, 22% developed borderline echocardiographic abnormalities. Six patients (22%) became symptomatic and four patients died of congestive heart failure. In all families cardiac abnormalities were found. There was no association between DCM and mutation type. Despite equal functional motor ability, there was a considerable intrafamilial variation in cardiac involvement, even in brother pairs. We conclude that cardiac abnormalities are the rule and not the exception in BMD and are progressive over time. Left ventricular dilatation may begin at any moment in the course of BMD and the rate of progression is unpredictable. A substantial proportion of patients will develop an incapacitating and life-threatening DCM. PMID- 9402543 TI - Slower recovery of muscle phosphocreatine in malignant hyperthermia-susceptible individuals assessed by 31P-MR spectroscopy. AB - Our aim was to develop an exercise protocol using 31P-magnetic resonance spectroscopy (31P-MRS), which can discriminate between malignant hyperthermia susceptible (MHS) individuals and controls. MRS spectra of the forearm muscles were recorded at rest, during and after a standardized exercise protocol in 10 MHS patients and compared with spectra obtained in 10 controls. There was no difference in resting intracellular pH (pHi) or PCr/(Pi+PCr) ratio between the groups (PCr = phosphocreatine, Pi = inorganic phosphorus). At the end of the exercise and during the initial recovery phase, the pHi and PCr/(Pi+PCr) ratio were significantly lower in the MHS group ([pHi: 6.37 (0.07) for MHS vs 6.70 (0.05) for controls, P < 0.005; PCr/(Pi+PCr): 0.784 (0.017) for MHS vs 0.954 (0.020) for controls, P < 0.0005]). For PCr/(Pi+PCr), complete separation between the two groups was observed during the initial recovery phase. The mean recovery time of PCr/(Pi+PCr) was 0.57 min for the control group and 1.28 min for the MHS group. The slower recovery of PCr/(Pi+PCr) is likely to be caused by a combination of several factors, including the lower pHi in MHS subjects at the start of recovery (inhibiting ATP production) and excessive sarcoplasmic calcium overload (causing continued enzyme activation and ATP consumption). Our exercise protocol can be a valuable adjunct to discriminate between MHS and non susceptible subjects. PMID- 9402545 TI - Slowly progressive isolated dysarthria: longitudinal course, speech features, and neuropsychological deficits. PMID- 9402546 TI - Vigabatrin-induced optic neuropathy. PMID- 9402547 TI - Palliative therapy in the terminal stage of neurological disease. AB - As recently pointed out by the American Academy of Neurology, providing adequate palliative care to dying patients is the duty of every neurologist. Because of a lack of relevant articles in the neurological literature, we have compiled current treatment recommendations for the most important symptoms arising in the endstage of neurological diseases. These recommendations include treatment of dyspnea, death rattle, restlessness, pain, thirst, depression, and others. A discussion of difficult decisions is included, e.g., the appropriate extent of fluid substitution or the ethical implications of sedation in the terminal phase. It is hoped that this compilation may provide a basis for future research in palliative therapy in neurology. PMID- 9402548 TI - Palliative care in amyotrophic lateral sclerosis. AB - The poor prognosis of amyotrophic lateral sclerosis (ALS) makes palliative care a challenge for the neurologist. Most disabilities associated with progressive disease can be ameliorated by symptomatic treatment. Prognosis and treatment options should be openly discussed with the patient and his/her relatives. Nutritional deficiency due to pronounced dysphagia can be efficiently relieved by a percutaneous enterogastrostomy. Respiratory insufficiency can be treated by non invasive ventilation at home, provided the familial environment is supportive. Adequate assistance and palliative treatment in the terminal phase is of paramount importance. PMID- 9402549 TI - Quality of life issues in palliative medicine. AB - The assessment of patient's quality of life is assuming increasing importance in medicine and health care. Illnesses, diseases and their treatments can have significant impacts on such areas of functioning as mobility, mood, life satisfaction, sexuality, cognition, and ability to fulfil occupational, social and family roles. The emerging quality of life construct may be viewed as a paradigm shift in outcome measurement since it shifts the focus of attention from symptoms to functioning. This holistic approach more clearly establishes the patient as the centre of attention and subsumes many of the traditional measures of outcome. Quality of life assessment is particularly relevant to patients with progressive conditions, particularly in the later phases of the disease. Despite the fact that current definitions of palliative medicine include quality of life as a central concern, relatively little research has been conducted on the impact of palliative care on patient quality of life. This paper introduces the concept of quality of life and describes the significant difficulties in definition, measurement and interpretation that must be addressed before such measures can be used as reliable and valid indicators of disease impact and treatment outcomes. It is argued that the unique individual perspective of the patient on his or her own quality of life must be incorporated into outcome assessments aimed at improving the quality of health care delivery in progressive diseases. PMID- 9402550 TI - Palliative medicine training for physicians. AB - In recent years, it has been widely recognised that modern doctors are not receiving the training they need to provide appropriate and effective palliative care. Deficiencies in care have been reported in several formal studies and, anecdotally, in many professional and popular lay publications. Doctors themselves have attested to their lack of confidence and competence in many aspects of palliative care. These problems cross national, cultural and religious boundaries, as evidenced by initiatives in many countries to correct the situation, which will be reviewed in this paper. PMID- 9402551 TI - The zeta pulse: a new stimulus waveform for use in electrical stimulation of the nervous system. AB - A new waveform which can be used instead of pulses for general electrical stimulation of the nervous system is described. This new waveform has been called the 'Zeta' pulse. The potential advantages of the use of this waveform are discussed together with a brief review and discussion of the mechanisms of extracellular stimulation in nervous tissue. A circuit diagram for the generation of the Zeta waveform is given. PMID- 9402552 TI - Visualization of significant differences in somatotopic maps: a distributed t test. AB - In order to test for differences in the properties of two populations of cells within a somatotopic map we need to be able to compare data sets in which sampled cells are randomly scattered throughout the map, and the variable being compared varies with location in the map. We can describe cell properties as exponentially smoothed surfaces fitted to data in the plane of the map, where all data contribute to the computation of the value of each grid point on the surface, with weights which decline exponentially with distance from the grid point. Means, variances and Student's t values can be computed at all grid points, keeping in mind the fact that grid points' t values are not independent of each other. We used Monte Carlo methods to demonstrate that two random samples of 500 values from two populations of 100,000 values at 4000 grid can provide a very useful picture of regions with significant differences. We recommended this procedure, or analogous approaches using other statistical tests, for any analysis where it is necessary to compare values of dependent variables when matched locations on the independent axis or plane cannot be sampled in the two populations. PMID- 9402553 TI - A new method for evaluation of motor deficits in 3-acetylpyridine-treated rats. AB - We established a novel method for quantitative evaluation of the motor deficits induced by 3-acetylpyridine (3-AP) in rats. 3-AP (75 mg/kg, i.p.) was injected in a time-controlled manner by a following injection of niacinamide (NIA; 300 mg/kg, i.p.). Changes in the motor function were evaluated by measuring the maximal height of vertical jump (MHVJ) to escape from an electrical shock on the foot, as well as ataxic gait. The MHVJ decreased and reached a minimum value 5-7 days after the 3-AP treatment. The close correlation of the MHVJ and the incidence of ataxic gait indicates that the decrease of MHVJ is a useful quantitative index of motor deficits. PMID- 9402554 TI - Development of a narrow water-immersion objective for laserinterferometric and electrophysiological applications in cell biology. AB - Laserinterferometric studies of the micromechanical properties of the organ of Corti using isolated temporal bone preparations are well established. However, there are relatively few measurements under in vivo conditions in the apical region of the cochlea because of its inaccessibility with commonly used techniques. Recently, optical-design programs have become affordable and powerful, so that the development of an optimized optical system is within the budget of physiologists and biophysicists. We describe here the development of a long-range water-immersion objective. To circumvent anatomical constraints, it has a narrow conical tip of taper 22 degrees and diameter 2.4 mm. It is a bright field reflected-light illumination, achromatic objective with magnification of 25x/infinity, a working distance of 2.180 mm and a numerical aperture of 0.45. Chromatic errors are corrected at 546.1 and 632.8 nm, with emphasis on the latter wavelength which is used by the laser interferometer. The field curvature is relatively flat and a diffraction limitation (Strehl ratio better than 0.8) can be obtained in a field of 0.4 mm diameter. Using this objective, sound-induced vibrations of hair cells and Hensen cells could be recorded without placing a reflector on the target area. In addition, this objective was found to be diffraction-limited in the near infra-red (750-830 nm), with a slightly different working distance (2.186 mm), making it suitable for patch-clamp experiments using infra-red, differential interference contrast. PMID- 9402555 TI - Isolation of single immunohistochemically identified whole neuronal cell bodies from post-mortem human brain for simultaneous analysis of multiple gene expression. AB - In Alzheimer's disease (AD), one cell in the brain may clearly be affected, while an adjacent cell appears healthy or unaffected. Previous technology has allowed us to examine one message at a time, at the level of a single cell (in situ hybridization, ISH), or multiple messages in a heterogeneous population of cells (Northern analysis). We have developed a methodology to build up a profile of multiple mRNA expression in single, whole, post-mortem cells that have been immunohistochemically (IHC) characterized. Fresh post-mortem tissue is spread into a layer one cell thick and fixed. Neurons are identified using an antibody to neurofilament and isolated using a micropipette. The mRNA is reverse transcribed and PCR carried out to confirm that material is present. A radioactively labeled antisense aRNA probe, which is representative of the messages contained in the cell is then amplified. This aRNA is used as a probe for a reverse Northern blot, allowing us to profile many genes from one cell at the same time. This technology has the potential to be applied to a wide variety of diseases encompassing many different cell types. PMID- 9402556 TI - A simple method, using 2-hydroxypropyl-beta-cyclodextrin, of administering alpha chloralose at room temperature. AB - Effective long term stable anaesthesia is a goal of many drug regimens employed in neuroscience in which procedures carried out are not practical in awake animals. A particular problem is the study of nociceptive mechanisms where good anaesthesia is essential. Similarly studies of cardiovascular or cerebrovascular mechanisms require that normal physiological reflexes be preserved as much as is practical. For non-recovery anaesthesia alpha-chloralose is a good choice since it provides good anaesthesia without excess depression of physiological reflexes. However, alpha-chloralose is sparingly soluble so that its use is not straightforward. We describe the characterisation of a simple procedure to solubilise alpha-chloralose in a solution of 2-hydroxypropyl-beta-cyclodextrin. The resulting solution is stable at room temperature and gives a high concentration of alpha-chloralose making it easier to administer regularly during longer time course experiments. PMID- 9402557 TI - A new protocol for the transmission of physiological signals by digital telemetry. AB - A method has been devised to transmit physiological signals from the brains of rats using a new digital telemetry transmission protocol. The chief advantage of the present system over existing systems is that the circuit only consumes power during the transmission of a brief pulse of information. This results in a very much extended battery life allowing the device to be implanted and recordings to be carried out over a longer period of time. PMID- 9402558 TI - Microelectrode arrays for stimulation of neural slice preparations. AB - A planar 6 x 6 array of iridium electrodes with four reference electrodes has been developed for use with neural tissue preparations. Precise knowledge of the relative locations of the array elements allows for spatial neurophysiological analyses. The 10 microns diameter platinized iridium electrodes on a 100 microns pitch have been used to stimulate acutely prepared slices of spinal cord from free-ranging rodents. An intracellular recording from a single neuron in the substantia gelatinosa (SG) using the whole-cell, tight-seal technique allowed low noise, high resolution studies of excitatory or inhibitory electrical responses of a given neuron to inputs from the primary afferent fibers or from stimulation by individual electrodes of the array. The resulting maps of responses provide an indication of the interconnectivity of neural processes. The pattern emerging is that of limited interconnectivity in the SG from areas surrounding a recorded neuron but with strong excitatory or inhibitory effects from those oriented in a longitudinal (rostral-caudal) direction relative to the neuron. The observations to date suggest the neurons of the SG are arranged in sets of independent networks, possibly related to sensory modality and input from particular body regions. PMID- 9402559 TI - Elvax as a slow-release delivery agent for a platelet-activating factor receptor agonist and antagonist. AB - The ability of the slow-release polymer, Elvax, to deliver a large number of different molecules to distinct brain regions has been well-documented. However, there are currently no reports of Elvax-mediated delivery of ether phospholipids, whose lipid structure renders them difficult to solubilize and detect under experimental conditions. In order to potentially examine the role of platelet activating factor (PAF) receptors in the brain in vivo, we tested the ability of Elvax to release the ether phospholipid. PAF receptor agonist, methyl-carbamyl platelet-activating factor (mc-PAF), and the ginkolide, PAF receptor antagonist, BN 52021. Mc-PAF and BN 52021 containing Elvax sections (200 microns) prepared in methylene chloride were assayed for release characteristics with a rabbit platelet aggregation bioassay. We measured a slow, sustained release of mc-PAF and BN 52021 from Elvax in vitro over a 5 day period. Similar in vivo mc-PAF and BN 52021 release kinetics were determined. Therefore, we believe that this novel method of slow-release delivery of PAF receptor agonists and antagonists as measured with platelet aggregation is viable and offers a simple, sensitive and inexpensive method for potential in vivo studies of the roles of PAF and its receptors in neural systems. PMID- 9402560 TI - Pseudo-immunolabelling with the avidin-biotin-peroxidase complex (ABC) due to the presence of endogenous biotin in retinal Muller cells of goldfish and salamander. AB - Immunodetection techniques are dependent on enzyme-protein conjugates for the visualisation of antigen-antibody complexes. One of the most widely used is the avidin-biotin-peroxidase complex (ABC) method. The present study demonstrates that direct treatment of goldfish and salamander retinal sections with ABC, followed by an incubation with the chromogenic substrate 3,3-diaminobenzidine tetrahydrochloride (DAB) and H2O2, manifested a punctate staining pattern across the neural retinae, presumably through binding of avidin to endogenous biotin. Incubation with a primary antiserum against biotin followed by immunoprocessing with the peroxidase--anti-peroxidase (PAP) method showed a pattern similar to the punctuate framework as detected with solo ABC-treated sections. Moreover, the ABC DAB/H2O2 mediated pattern corresponded to the spatial orientation of Muller cells as identified by GFAP immunostaining. These findings indicate the presence of endogenous biotin in Muller cells and calls for caution in the application of the ABC method in immunotechniques in retinal research. PMID- 9402561 TI - Multivariate analysis of RNA levels from postmortem human brains as measured by three different methods of RT-PCR. Stanley Neuropathology Consortium. AB - The analysis of RNA from postmortem human brain tissue by reverse transcription polymerase chain reaction (RT-PCR) provides a practical method to measure both normal and abnormal brain gene expression. A major limitation in using human material is that yields can vary dramatically from individual to individual, making comparisons between samples difficult. In this report, we study the association of pH and several pre- and postmortem factors on the RNA yields from 89 postmortem human occipital cortices. Glyceraldehyde phosphate dehydrogenase (GAPdH) mRNA levels were measured by RT-PCR. A major variant in this method is the priming used in the reverse transcription reaction. Three different methods of reverse transcription were performed and the resultant levels of products compared against the pre- and postmortem factors and pH. The levels of GAPdH correlated significantly to pH and pH itself to the rapidity of death (RoD) (agonal state) indicating that premortem factors may play the greatest role in determining postmortem RNA levels. The three methods of priming showed different sensitivities, most notably that oligo dT priming alone is vulnerable to long freezer intervals (FI). We conclude that premortem factors are the major affectors of RNA levels variations and that the polyA tail region of the molecule appears to be adversely affected by extended freezer storage. PMID- 9402562 TI - Identification of synaptic connections in neural ensembles by graphical models. AB - A method for the identification of direct synaptic connections in a larger neural net is presented. It is based on a conditional correlation graph for multivariate point processes. The connections are identified via the partial spectral coherence of two neurons, given all others. It is shown how these coherences can be calculated by inversion of the spectral density matrix. In simulations with GENESIS, we discuss the relevance of the method for identifying different neural ensembles including an excitatory feedback loop and networks with lateral inhibitions. PMID- 9402563 TI - Generation of anti-peptide antibodies against serotonin 5-HT2A and 5-HT2C receptors. AB - Anti-peptide antibodies were generated against several 13-17 amino acid regions of rat serotonin 5-HT2A and 5-HT2C receptors. Peptides containing terminal cysteine residues were conjugated to bovine serum albumin (BSA) and ovalbumin (OVA) with the cross-linking reagent sulfo-SMCC (sulfosuccinimidyl 4-(N maleimidomethyl) cyclohexane-1-carboxylate). Both the carrier protein and the number of peptide molecules per carrier molecule were changed during the immunization schedule. For the early immunizations, immunogens were BSA-peptides at ratios of 8-27 mol peptide per mol BSA. For the later boosts, immunogens were OVA-peptides at ratios of 1-2 mol peptide per mol OVA. The peptide constructs were used to immunize rabbits and chickens. Anti-peptide antibodies were purified from sera (rabbits) or egg yolks (hens) using peptide matrices. Cell lines expressing similar densities of rat 5-HT2A or 5-HT2C receptors were used to monitor the specificity of purified antibodies on immunoblots and in immunocytochemistry. A total of five out of the six rabbit antibodies were positive on immunoblots (three anti-5-HT2A and two anti-5-HT2C) and four were also positive in immunocytochemistry (three anti-5-HT2A and one anti-5-HT2C). None of the anti-peptide chicken antibodies were useful on immunoblots or in immunocytochemistry. Since there is a paucity of high affinity reagents selective for 5-HT2A or 5-HT2C receptors, these rabbit antibodies will be useful tools. The methods used to generate site-directed antibodies specific for 5-HT2A or 5-HT2C receptors should be applicable to other proteins. PMID- 9402564 TI - The effects of patellar taping on stride characteristics and joint motion in subjects with patellofemoral pain. AB - Although patellar taping has been reported to be effective in reducing pain, the effects of this procedure on functional outcomes, such as ambulation, have not been documented. The purpose of this study was to compare stride characteristics and joint motion in subjects with patellofemoral pain, with and without the application of patellar taping using the McConnell technique. Fifteen female subjects between the ages of 14 and 41 years with diagnosis of patellofemoral pain participated in this study. Stride characteristics (Stride Analyzer) and sagittal plane joint motion (VICON) were recorded simultaneously during taped and untaped trials of free walking, fast walking, and ascending and descending a ramp and stairs. A repeated measures analysis of variance was used to determine differences between taped and untaped trials. Although subjects reported an average pain reduction of 78% using a visual analogue scale, the only significant change in stride characteristics was an increase in stride length during ramp ascent. Patellar taping did, however, result in a small but significant increase in loading response knee flexion across all conditions tested. We believe this finding demonstrates more willingness by the patellofemoral pain subjects to load the knee joint, thus permitting increased shock absorption, increased quadriceps activity, and tolerance of increased patellofemoral joint reaction force. PMID- 9402565 TI - Electromyographic analysis of selected lower extremity musculature in normal subjects during ambulation with and without a Protonics knee brace. AB - Often, braces are an integral part of treatment programs for patients with pathology of the knee joint. Little evidence exists, however, as to the effect of braces on muscle function. The purpose of this investigation was to compare electromyography (EMG) from six lower extremity muscles during level walking without the Protonics knee brace and with the brace at eight resistance settings. Surface electrodes were placed on one lower extremity of 19 subjects (ages = 21 57) to evaluate EMG activity during ambulation with and without the knee brace. Data were normalized to maximum voluntary contractions and averaged across cycles. There was a significant increase in muscle activity of the rectus femoris, vastus medialis, and vastus lateralis muscles when the brace resisted knee extension and was set at the level of 9. Significantly higher EMG levels also occurred in the vastus lateralis and vastus medialis with the extension module set at level 6 when compared with the no brace trial and resistance levels set at 6 and 2 with the flexion module. In this normal population, there was an increase in activity of selected muscles when the brace was set at the highest resistance settings. These data serve as a guide for clinicians when considering incorporation of a brace of this type into patient management. PMID- 9402566 TI - The three-dimensional passive support characteristics of ankle braces. AB - Studies of the passive support provided by ankle braces have focused primarily on inversion support. The goal of this study was to develop a technique to measure the support provided by ankle braces in all rotational directions and to use this technique to compare four common braces (Ascend, Swede-O, Aircast, and Active Ankle). For this purpose, a 6 degrees-of-freedom linkage was used to measure the flexibility of the ankle complex in 10 healthy subjects. Each subject was tested without brace support and with each of the four braces. Testing was repeated on each subject on two different occasions. The angular displacement at specified moment values and the four segmental flexibility values obtained from the loading portion of the moment-angular displacement data were used in the data analysis. Repeated measure analysis of variance followed by a Student Neuman-Keuls test at p < 0.05 was performed. This statistical analysis was used to identify significant differences among the braces and differences between each brace and the no brace condition. Each of the four braces provided significant support in inversion, eversion, and internal rotation, but the amount of support varied significantly among the braces. In external rotation, only the stirrup braces provided significant support. The braces also varied significantly in the amount of interference with dorsiflexion and plantar flexion. Clinicians may be assisted by objective data on the amount and nature of passive support when prescribing braces to their patients. PMID- 9402567 TI - The slump test: the effects of head and lower extremity position on knee extension. AB - Maitland's slump test is a widely used neural tissue tension test. During slump testing, terminal knee extension is assessed for signs of restricted range of motion (ROM), which may indicate impaired neural tissue mobility. A number of refinements that modify hip and ankle position has been added to the basic slump test procedure, but no research to date has measured the effects of ankle and hip position on knee extension ROM during testing. The purpose of this study was to examine the effect of neural tension-producing movements of the cervical spine and lower extremity on knee extension ROM during the slump test. Thirty-four males with no significant history of low back pain were tested in the slump position with the cervical spine flexed and extended in each of three lower extremity test positions: neutral hip rotation with the ankle in a position of subject comfort (neutral), neutral hip rotation with ankle dorsiflexion (ankle dorsiflexion), and medial hip rotation with ankle dorsiflexion. Results showed significant decreases in active knee extension ROM (F1,198 = 29.53, p < 0.0001) in the cervical flexion compared with the cervical extension conditions. Subjects also exhibited significant decreases in active knee extension ROM (F2,198 = 56.76, p < 0.0001) as they were progressed from neutral to the ankle dorsiflexion to the medial hip rotation with ankle dorsiflexion positions of the lower extremity. The results of our study indicate that limitations in terminal knee extension ROM may be considered a normal response to the inclusion of cervical flexion, ankle dorsiflexion, or medial hip rotation in the slump test in young, healthy, adult males. In addition, the presence of a cumulative effect on knee extension ROM with the simultaneous application of these motions is noted. These findings may assist clinicians when assessing knee extension ROM during slump testing. PMID- 9402568 TI - Effectiveness of visual feedback during isokinetic exercise. AB - Although previous investigators have observed that knowledge of performance via visual feedback tends to enhance performance during an isokinetic test, the time frame over which visual feedback remains advantageous is unclear. The purpose of this study was to compare knee extensor torques produced by visual feedback and no visual feedback groups on three occasions, completed over a 2-week period, and at 4 weeks after the third test. Healthy, sedentary subjects were each randomly assigned to either a visual feedback or a no visual feedback group (N = 10 males and 10 females per group). Visual feedback consisted of viewing a computer monitor which displayed the current and a target knee extension force. Torques produced by the visual feedback group were consistently greater (p < 0.05) and more reliable than those produced by the no visual feedback group. The effectiveness of visual feedback tended to decrease over the first three occasions, suggesting that visual feedback may not be as advantageous once a skill is well learned. Further research needs to examine the contribution of visual feedback to motor learning as well as retention and transfer of motor skills during more complex functional tasks. PMID- 9402569 TI - Assessing anterior cruciate ligament injuries: the association and differential value of questionnaires, clinical tests, and functional tests. AB - It is important to examine the package of questionnaires and clinical and functional tests as used in anterior cruciate ligament (ACL)-injured patients in order to gain insight on the patient's present status. Nine measuring systems in three categories were examined: four questionnaires, three clinical tests, and two functional tests. Differences between sports activity rating system, factor occupational rating system scale, and Tegner scores pre- and post-injury and the differences between the affected and unaffected knee in the clinical and functional tests were calculated using the Wilcoxon test for paired observations. These differences proved to be significant (p < 0.05). The association between the various tests was also examined. None of the associations satisfied the preset standards. Based on these low levels of association, it does not seem possible to reduce the package of tests to one questionnaire, one clinical test, and one functional test as all questionnaires and tests seem to be related to different aspects of the injured ACL. Based upon these results, the total package should be used to gain insight in both impairment and disability level in patients with an injured ACL. PMID- 9402570 TI - The effects of the number and frequency of physical therapy treatments on selected outcomes of treatment in patients with anterior cruciate ligament reconstruction. AB - Health care reform will quite possibly change the delivery of physical therapy by demanding physical therapists to be more accountable for providing appropriate, yet cost-effective treatment. The purpose of this study was to retrospectively compare the results after anterior cruciate ligament (ACL) reconstruction between two groups of patients with different numbers and frequencies of physical therapy visits postoperatively. Two random samples of 100 patients from a total of 1,345 patients identified as undergoing ACL reconstruction from 1990 through 1993 were included. Group A patients attended physical therapy regularly and participated in a home exercise program, while patients in Group B attended limited physical therapy visits and also performed a prescribed home exercise program. Both groups followed the same postoperative rehabilitation program for early range of motion, early weight bearing, and muscle control. The outcome variables measured 1, 6, and 12 months postoperatively included the number of structured visits to physical therapy, range of motion, isokinetic strength testing, and subjective rating. Group A averaged 20 visits in the first 6 months while Group B averaged seven visits. The results revealed no significant difference for flexion, isokinetic strength, or subjective rating. There was a significant difference for hyperextension (Group A, 2 degrees; Group B, 6 degrees). The results of this investigation indicated that by following a structured physical therapy program postoperatively, it is possible for patients to achieve a successful outcome with a limited number of routine physical therapy visits. PMID- 9402571 TI - Effects of detraining on knee extensor strength and functional mobility in a group of elderly women. AB - Long-term detraining results for individuals 75 years and older are needed. The purpose of this study was to assess long-term detraining effects on quadriceps strength and functional mobility in nursing home residents. Ten women (X = 82.8 years) who completed a strength training program were reassessed 1 year later. Clinical methods were used to remeasure dynamic and isometric quadriceps strength and functional mobility. One repetition maximum quadriceps strength declined 68.3% (p < 0.05) from trained values. Isometric strength losses were 29.8% at 90 degrees (p < 0.05), 28.7% at 60 degrees (p < 0.05), and 24.4% at 20 degrees (p < 0.05) of knee flexion 1 year postexercise. Fast-paced walking, self-selected paced walking, and timed up and go speed decreased 28.6% (p < 0.05), 19.5% (p < 0.05), and 54.1% (not significant), respectively, from posttraining. One year vs. baseline, isometric strength decreased 0-14.3%, dynamic strength decreased 48.9%, and functional mobility declined 16.5-20.7% despite an intervening training program. An increased strength loss rate beyond the age of 80 years may be a major factor influencing functional independence. PMID- 9402572 TI - Management of shoulder dysfunction with an alternative model of orthopaedic physical therapy intervention: a case report. AB - One common approach to patient care in dealing with many musculoskeletal dysfunctions involves two to three patient visits to physical therapy per week over a period of weeks. Some patients may benefit from an alternative, graduated treatment model emphasizing a minimal number of office visits and focusing on intensive patient education, home program therapeutic exercise, and specific manual interventions. Patient education focuses on home program compliance and empowerment of the patient by adjusting office visits as needed based on patient progress rather than multiple patient contacts in the first weeks. This emphasis may improve long-term patient compliance by preventing the development of an external locus of control in which the patient is dependent upon the therapist for management of his/her condition. This case study is an example of the use of this alternative treatment model for the resolution of impingement syndrome and adhesive capsulitis in a 53-year-old female. A comprehensive program of patient education and home exercise was initiated during the first visit. Joint mobilization and active exercise were performed at each subsequent visit. The patient was seen a total of six visits over a period of approximately 10 1/2 weeks, followed up via telephone at 1 month after the last treatment and reexamined after 1 year. The objective exam revealed no abnormalities after the last visit or after 1 year. The patient subjectively reported compliance with the home program for 6 months after the last visit. This model of patient care was successful for the patient described in this case study. The treatment approach may have contributed to the development of an internal locus of control by allowing the patient to be as actively involved as possible in the treatment of her condition. In addition, this approach is timely when one considers current reimbursement systems. Though successful with this patient, this graduated treatment model is not intended to be applicable to every patient with this diagnosis. PMID- 9402573 TI - Treatment of inoperable carotid aneurysms with endovascular carotid occlusion after extracranial-intracranial bypass surgery. AB - OBJECTIVE: Hunterian ligation of the internal carotid artery (ICA) is an accepted treatment for inoperable carotid aneurysms. Preliminary extracranial-intracranial (EC-IC) bypass surgery is required in some patients. The reported incidence of thromboembolic and ischemic complications remains significant for these patients, despite a variety of advocated management strategies. We present our treatment paradigm. METHODS: Between April 1992 and March 1997, nine patients with inoperable ICA aneurysms were treated using EC-IC bypass surgery and then permanent endovascular ICA occlusion. All of the patients except one had been selected for bypass surgery on the basis of failing results of the ICA test occlusion with hypotensive challenge. ICA occlusion was performed by endovascular means and was delayed after bypass surgery was performed by a mean of 6 days (range, 2-20 d). All patients were managed in the intensive care unit after ICA occlusion. RESULTS: Clinical improvement was noted in all patients (mean follow up, 21 mo; range, 3-42 mo). There were no major complications. Aneurysmal thrombosis was confirmed in all patients. Although ICA occlusion was delayed after bypass surgery, only one bypass was noted to be occluded. The occluded bypass occurred in a patient who subsequently underwent successful ICA occlusion. This patient was thought to have been improperly selected for bypass surgery. CONCLUSION: Certain carotid aneurysms can be effectively managed with hunterian ICA ligation. After preliminary identification of patients with borderline cerebrovascular reserve as candidates for EC-IC bypass surgery, close attention to the following points may help enhance clinical outcome: 1) excellence in surgical technique for EC-IC bypass surgery, 2) occlusion of the parent vessel as close to the aneurysm neck as possible by endovascular means, and 3) judicious postoperative combination of anticoagulation, fluid, and pressure management. PMID- 9402574 TI - Safety and efficacy of endovascular treatment of acutely ruptured aneurysms. AB - OBJECTIVE: To study the safety and efficacy of endovascular treatment of acutely ruptured aneurysms with Guglielmi detachable coils. METHODS: From August 1992 until December 1995, 75 patients were referred for endovascular treatment of acutely ruptured aneurysms. There were 49 women and 26 men, with a mean age of 55 years. Patients were classified according to the Hunt and Hess grading system. There were 18 Grade I patients (24%), 13 Grade II patients (17%), 30 Grade III patients (40%), 11 Grade IV patients (15%), and 3 Grade V patients (4%). Fifty patients (66%) were treated within 48 hours, and 64 (85%) were treated within 1 week of hemorrhage. The most frequently treated aneurysms were located at the basilar bifurcation (32%), anterior communicating artery (16%), posterior communicating artery (15%), and ophthalmic segment of the carotid artery (11%). Most of the aneurysms were smaller than 15 mm (77%). Fifty-six percent of the aneurysms had small (4 mm) necks, and 44% had wide (> 4 mm) necks. Clinical follow-up was performed at 6 months, and results were classified according to the Glasgow Outcome Scale (GOS). Control angiograms were performed immediately, at 6 months, and yearly thereafter. RESULTS: Immediate angiographic results were considered to be satisfactory in 58 patients (77%) (complete obliteration, 40%; residual neck and dog ear, 37%). Technical failures occurred in 5 patients (7%), and 12 patients experienced some residual opacification of their aneurysms (16%). The procedure-related mortality and morbidity rate was 8%. At 6 months, the outcomes were as follows: GOS score of 1, 50 patients (66.7%); GOS score of 2, 4 patients (5.3%); GOS score of 3, 4 patients (5.3%); and GOS score of 5, 17 patients (22.7%). The main causes of death and disability at 6 months were the direct effect of the initial hemorrhage (9%), delayed ischemia (6.7%), subsequent bleeding (4%), intraprocedural rupture (4%), open surgical complications (3%), and unrelated deaths (4%). Six-month angiographic follow-up data were available for 50 patients (67%). The morphological results were considered to be satisfactory in 44 of these 50 patients (88%) (complete occlusion, 46%; residual neck or dog ear, 42%). CONCLUSION: Endovascular treatment of acutely ruptured aneurysms was attempted without clinically significant complication in 92% of the patients. The morphological results were unsatisfactory in 23% of the patients. Complete obliteration of the sac, with or without residual neck, is essential to prevent subsequent bleeding, which occurred in 5% of the patients. The overall outcome at 6 months was similar to that of surgical series, despite a selected group of patients with negative prognostic factors. PMID- 9402575 TI - Familial intracranial aneurysms: an autopsy study. AB - OBJECTIVE: Familial intracranial aneurysms are more common than has been appreciated, but systematic autopsy studies of affected individuals have not been reported. We reviewed the autopsy findings of a group of patients with familial aneurysms to elucidate the nature of the putative underlying arteriopathy. METHODS: Using a computerized diagnostic index, we identified all patients with intracranial aneurysms in whom postmortem examination had been performed at the Mayo Clinic between January 1, 1992, and December 31, 1994. The medical records, radiographic studies, and autopsy findings of these patients were reviewed. RESULTS: Among the 28 patients with intracranial aneurysms, 3 (11%) had one or more first-degree relatives with documented intracranial aneurysms. The mean age of the three patients (two women and one man) was 54 years. Microscopic examination of the vascular system revealed medial changes, consisting of degeneration of elastic fibers and increased ground substance, in the systemic arteries of 2 of the 3 patients with familial aneurysms but in none of the 25 patients with sporadic aneurysms. These nonspecific medial changes involved both common and extracranial internal carotid arteries in one patient and the entire aorta as well as intracranial and common carotid arteries in the other patient. CONCLUSION: These observations suggest that an underlying arteriopathy in patients with familial intracranial aneurysms involves the tunica media and commonly may affect systemic (extracranial) arteries. PMID- 9402576 TI - Preoperative activation and intraoperative stimulation of language-related areas in patients with glioma. AB - OBJECTIVE: Evaluation of the accuracy of preoperative localization of language related cortex by magnetic resonance imaging-guided positron emission tomography. METHODS: Patients with gliomas in the left dominant hemisphere were examined preoperatively with magnetic resonance imaging-guided positron emission tomography and intraoperatively by electrical stimulation of cortex. RESULTS: A verb generation task yielded more intense and better lateralized local increases of cerebral blood flow in the positron emission tomographic examination than did a naming task. Significant correspondence of preoperative and intraoperative findings was observed for the verb generation task. Cortical sites with aphasic disturbance during electrical stimulation had a significantly higher cerebral blood flow increase during preoperative activation than did sites without intraoperative language impairment. Areas with cerebral blood flow increases above an optimum threshold had 73% sensitivity and 81% specificity to predict aphasic disturbance during intraoperative stimulation. CONCLUSION: The data suggest that with further technical improvements, imaging of language function may become a preoperative diagnostic tool for patients with tumors close to language-related brain structures. PMID- 9402577 TI - Infratentorial empyema: analysis of 22 cases. AB - OBJECTIVE: Infratentorial empyema is an uncommon form of intracranial suppuration that is usually secondary to neglected otogenic infection. The diagnosis is frequently delayed and often confused with that of meningitis. The associated mortality is distressingly high, yet it has, as a clinical entity, received scant attention in the literature. We present a 13-year experience of this condition. PATIENTS AND METHODS: From a retrospective analysis of 3865 patients with intracranial suppuration during a 13-year period, 22 patients with infratentorial empyema were identified. The inpatient notes for these patients were analyzed with reference to clinical, radiological, bacteriological, operative, and outcome data. RESULTS: Twenty-two patients with infratentorial empyema accounted for 0.6% of admissions caused by intracranial suppuration during the study period. Of these 22 empyemas, 13 were subdural and 9 epidural. Hydrocephalus was present in 17 (77.3%). Except for two epidural empyemas that did not warrant neurosurgical intervention, all patients underwent standard surgical management (wide posterior fossa craniectomy). Nineteen underwent mastoidectomy because the source of infection was otogenic. Concomitant and persistent hydrocephalus was treated aggressively. Five patients died (mortality rate of 22.7%). All fatalities had subdural empyemas, and all three patients with cerebellopontine angle extension of subdural purulent collections died. CONCLUSION: Although rare, infratentorial empyema, especially when subdural, is a lethal disease. Cerebellopontine angle extension of pus was a particularly ominous sign in our experience. Early surgical drainage via wide posterior fossa craniectomy, aggressive treatment of associated hydrocephalus, eradication of the primary source of sepsis, and, finally, intravenous high dosage of appropriate antibiotics form the mainstay of treatment. PMID- 9402578 TI - Current treatment of brain abscess in patients with congenital cyanotic heart disease. AB - OBJECTIVE: The goal of this study was to define clearly the role of management in patients with cyanotic heart disease and brain abscesses by evaluating retrospectively the factors influencing poor outcome in these patients. METHODS: This study included 62 patients with cyanotic heart disease and brain abscesses diagnosed in the computed tomography era. Basic characteristic parameters (number, size, location, computed tomographic classification and organism type of abscess, convulsion, type of cyanotic heart disease, age distribution, immunocompromised status, pretreatment neurological state, and intraventricular rupture of brain abscess [IVROBA]) and therapeutic parameters (type of antibiotics and duration of administration, steroid medication and therapeutic modalities, aspiration with or without cerebrospinal fluid drainage, total extirpation after aspiration, or primary extirpation and medical treatment) were evaluated as independent predictors of poor outcome (totally disabled state or death) by using univariate and multivariate logistic regression analysis. We also statistically estimated the possible causes of IVROBA and the multiplicity of brain abscess. RESULTS: Although there were no statistically significant correlations between patients with good and poor outcomes in regard to other basic characteristic and therapeutic parameters, patients with poor outcomes were older (P < 0.02), more frequently had IVROBA (P < 0.005), and had a higher frequency of neurological deterioration (P < 0.01) than those with good outcomes. Multiple logistic regression analysis predicted that poor outcome increased the relative risk of IVROBA by a factor of 18.9 (odds rate, 18.9; 95% confidence interval, 1.7-211.6; P < 0.02). More patients with multiple abscesses had positive immunocompromised states than those with single abscesses (P < 0.01). Deep-located abscesses also more frequently had IVROBA (P < 0.005) and abscesses located in the parieto-occipital region ruptured into the occipital horn of the lateral ventricle in a short period (P < 0.02). CONCLUSIONS: Our findings suggest that IVROBA strongly influences poor outcome in patients with cyanotic heart disease. The key to decreasing poor outcomes may be the prevention and management of IVROBA. To reduce operative and anesthetic risk in these patients, abscesses should be managed by less invasive aspiration methods guided by computed tomography. Abscesses larger than 2 cm in diameter, in deep-located or parieto occipital regions, should be aspirated immediately and repeatedly, mainly using computed tomography-guided methods to decrease intracranial pressure and avoid IVROBA. IVROBA should be aggressively treated by aspiration methods for the abscess coupled with the appropriate intravenous and intrathecal administration of antibiotics while evaluating intracranial pressure pathophysiology. PMID- 9402579 TI - Use of cerebrospinal fluid shunts in patients having acquired immunodeficiency syndrome with cryptococcal meningitis and uncontrollable intracranial hypertension. AB - OBJECTIVE: To evaluate the treatment of serious and uncontrollable intracranial hypertension in patients with acquired immunodeficiency syndrome who developed cryptococcal meningitis. METHODS: All cases of cryptococcal meningitis with elevated pressure and acquired immunodeficiency syndrome were reviewed in detail and described. RESULTS: Cerebrospinal fluid shunting dramatically improved these critically ill patients and was much more successful than serial lumbar punctures or the use of high-dose dexamethasone. CONCLUSION: Patients with acquired immunodeficiency syndrome who develop cryptococcal meningitis and who suffer serious visual loss or ocular palsies with elevated pressures should be considered for cerebrospinal fluid shunting at an early stage. PMID- 9402581 TI - Recent advances in epilepsy surgery: temporal lobectomy and multiple subpial transections. AB - THIS ARTICLE REVIEWS four major advances in epilepsy surgery, especially the most frequently performed surgery, temporal lobectomy, as follows: 1) the ability to preoperatively identify (using magnetic resonance imaging) the pathological condition of hippocampal sclerosis (a key component to the syndrome of mesial temporal lobe epilepsy, 2) the ability to identify preoperatively which temporal lobe candidates are at risk for postoperative memory problems, 3) the standardization of temporal lobectomy with respect to how much hippocampus should be resected, 4) a validation of the novel surgical technique of multiple subpial transections. This technique allows surgeons to attack foci within nondispensible cortex and therefore enlarges the applicability of surgical treatment to otherwise inoperable patients and potentially improves outcome. PMID- 9402580 TI - Cerebrospinal fluid adenosine concentration and uncoupling of cerebral blood flow and oxidative metabolism after severe head injury in humans. AB - OBJECTIVE: Uncoupling of cerebral blood flow (CBF) and oxidative metabolism is observed after severe head injury in comatose patients; however, the mechanism(s) involved remain undefined. Adenosine can produce cerebral vasodilation and reduce neuronal activity and is a possible mediator of uncoupling. We hypothesized that cerebrospinal fluid (CSF) adenosine concentrations would be increased during uncoupling of CBF and oxidative metabolism, defined as a narrow arterio-jugular venous oxygen difference [D(a-v)O2 4 vol%] after head injury. METHODS: Adenosine concentrations were measured using fluorescent-based high-pressure liquid chromatography in 67 CSF samples obtained from 13 comatose (Glasgow Coma Scale score 7) adult patients who sustained a severe closed head injury. At the time each sample was obtained, CBF was measured by the xenon-133 method, and blood samples were obtained for determination of D(a-v)O2. RESULTS: CSF adenosine concentration was negatively associated with D(a-v)O2 (P < 0.05, generalized multivariate linear regression model). In addition, CSF adenosine concentration was increased when D(a-v)O2 was 4 versus > 4 vol% (38.5 [3.2-306.3] versus 14.0 [2.7-795.5] nmol/L, respectively, median [range]; P < 0.025) and in patients who died versus survivors (40.1 [6.9-306.3] versus 12.9 [2.7-795.5] nmol/L, respectively, median [range]; P < 0.001). CONCLUSION: The association between increased CSF adenosine concentration and a reduction in global cross-brain extraction of oxygen supports a regulatory role for adenosine in the complex balance between CBF and oxidative and nonoxidative metabolism severe head injury in humans. PMID- 9402582 TI - Analysis of pallidotomy lesion positions using three-dimensional reconstruction of pallidal lesions, the basal ganglia, and the optic tract. AB - OBJECTIVE: To assess the position of radiofrequency pallidotomy lesions placed using microelectrode stimulation and cellular recordings in relation to a stereotactically defined starting point. Radiofrequency lesion locations were also evaluated in relation to the putamen, posterior limb of the internal capsule, and optic tract. METHODS: Magnetic resonance images obtained from 23 patients with Parkinson's disease who underwent pallidotomy at the University of Kansas Medical Center were analyzed. Using computerized techniques, lesion positions in relation to the midcommissural point and a hypothetical starting point were determined. Data segmentation and three-dimensional reconstruction of pallidal lesions, the internal capsule, and the optic tract allowed assessment of lesion position in relation to internal anatomy. Clinical outcome of pallidotomy was assessed using both the Unified Parkinson's Disease Rating Scale and the Dementia Rating Scale. RESULTS: Pallidal lesions were usually placed anterior and dorsal to the stereotactically defined starting point. The position of pallidal lesions in the men were observed, in four trials, to be significantly more dorsal than the lesions in the women. The outer zone of the lesion was usually adjacent to the internal capsule and the putamen and relatively close to the optic tract. The inner zone of the lesion was usually several millimeters removed from anatomic boundaries of the putamen, internal capsule, and optic tract. Patients achieved favorable outcomes, with reduced dyskinesias and "off" time and improvement of their Parkinsonian symptoms, as evidenced by clinical assessment, the Unified Parkinson's Disease Rating Scale, and the Dementia Rating Scale. CONCLUSION: Microelectrode stimulation and cellular recordings usually led to a final pallidotomy lesion position that deviated from the stereotactically defined starting point. The pallidotomy lesions in the men were observed to be more dorsal than the lesions in the women. Clinical outcomes were not correlated with either lesion location relative to the starting point or distances between the pallidal lesion and the putamen, internal capsule, or optic tract. Kinesthetically responsive cells may be localized generally more anterior and dorsal to the starting point (within the globus pallidus) and may be grouped variably from patient to patient in relation to other basal ganglia structures. Although the primary lesion site is most likely within the sensorimotor region of the globus pallidus internus, the more dorsal locations of responsive cell groups may indicate that some lesion sites may be localized within the globus pallidus externus. PMID- 9402583 TI - Stereotactic transcranial magnetic stimulation: correlation with direct electrical cortical stimulation. AB - OBJECTIVE: To evaluate stereotactic transcranial magnetic stimulation (TMS) as a tool for presurgical functional mapping of human motor cortex. METHODS: Transcranial magnetic stimulation using a frameless stereotactic system was performed in two patients with tumors near the central sulcus. TMS motor function maps were plotted on the patients' three-dimensional volumetric magnetic resonance imaging data and compared with direct electrical cortical stimulation at surgery with the patient under local anesthesia. RESULTS: Stereotactic TMS was well tolerated by both patients and was consistent with known somatotopic representation of human motor cortex. The results demonstrated a good correlation between the TMS and electrical cortical stimulation maps, with all TMS responses eliciting more than 75% of the maximum motor evoked potential falling within 1 cm of the electrical cortical stimulation site. CONCLUSIONS: Our findings indicate that stereotactic TMS is feasible and can provide accurate noninvasive localization of cortical motor function. It may prove to be a useful method for presurgical planning. PMID- 9402584 TI - The role of motor evoked potentials during surgery for intramedullary spinal cord tumors. AB - OBJECTIVE: This is a prospective study of the methodology and clinical applications of motor evoked potentials (MEPs) during surgery for intramedullary spinal cord tumors. METHODS: Transcranial electrical stimulation was used to activate corticospinal motoneurons, and the traveling waves of the spinal cord were recorded through catheter-electrodes placed epi- or subdurally. Intraoperative MEP monitoring was performed in 32 consecutive patients (age range, 1-50 yr) undergoing resection of intramedullary spinal cord tumors. In 19 patients, MEPs were present before myelotomy (monitorable group), and in 10 patients, MEPs were absent before myelotomy (unmonitorable group). Placement of an epidural electrode was not possible in two patients, and technical problems prevented recording in one. RESULTS: MEP amplitudes decreased intraoperatively by more than 50% of baseline in three patients, all of whom had postoperative paraplegia. Two of these patients recovered within 1 week after surgery, and one remained paraplegic. None of the patients with preserved MEP amplitude (> 50%) sustained immediate significant postoperative deterioration. Motor function was significantly deteriorated 1 week after surgery in one patient in the monitorable group and in five patients in the unmonitorable group. MEP monitorability was significantly associated with good surgical outcome for adult patients (P < 0.05), although not for pediatric patients (P > 0.6). Preoperative motor status and surgical outcome were not significantly associated for the adult (P = 0.13) or pediatric groups (P > 0.4). CONCLUSION: MEP monitorability was a better predictor of functional outcome than the patient's preoperative motor status for the adult group. Significant predictors of MEP monitorability in the adult group were preoperative motor function (P < 0.01), history of no previous treatment (surgery or irradiation) (P < 0.01), and small tumor size (P < 0.05). Weak associations with monitorable MEPs existed for low-grade tumors (P = 0.09), the presence of baseline somatosensory evoked potentials (P = 0.10), and tumor pathological abnormalities (ependymoma) (P = 0.13). No associations were determined for sex (P > 0.4), associated syrinx (P > 0.3), or tumor location (P > 0.5). In the pediatric group, none of the examined factors were associated with MEP monitorability (P > 0.3). A decline of more than 50% in MEP amplitude during tumor removal should serve as a serious warning sign to the surgeon. PMID- 9402585 TI - Value of nerve action potentials in the surgical management of traumatic nerve lesions. AB - OBJECTIVE: The goals of the study were to investigate the value of intraoperative electrically evoked nerve action potentials (NAPs) in the surgical treatment of traumatic peripheral nerve injuries (nerve lesions in continuity). METHODS: Sixty four patients with 76 traumatic nerve lesions in continuity were investigated intraoperatively by stimulating and recording NAP from the whole nerve across the suspected lesion site. Among the 76 nerves (nerve lesions) were 43 with incomplete and 33 with complete loss of function. In cases (nerves) with complete loss of function (n = 33), the surgical procedure (external neurolysis, internal neurolysis, or nerve repair) was performed according to the microscopic aspect of the nerve and the result of the intraoperative electrophysiological testing. In cases (nerves) with incomplete loss of function (n = 43), the surgical procedure was performed solely according to the microscopic aspect of the nerve and independently from the result of the intraoperative electrophysiological testing. RESULTS: Of 43 nerves with incomplete loss of function, we were able to record reproducible NAPs in 41 (95%) across the lesion site, thus demonstrating a high reliability of the method. Of 33 nerves with complete loss of function, a reproducible NAP could be recorded only in 3. Assuming an axonotmetic lesion in regeneration, we did nothing else on the nerve with excellent clinical results (full recovery). Of the remaining nerves with no NAP, 24 showed a caliber shift of the nerve (in 20 cases a thickening of the nerve, suggesting a neuroma in continuity). A grafting procedure was performed, and the histological evaluation revealed a neurotmetic lesion. However, in six patients with no NAP, there was no clear caliber shift of the nerve. The epineurium was opened and an internal neurolysis performed showing fascicles in continuity. Three patients had good and three had partial (but useful) recovery. CONCLUSIONS: In nerve lesions in continuity with complete loss of nerve function, intraoperative NAPs are able to detect axonotmetic lesions in regeneration. Thus, unnecessary further surgical procedures can be avoided. On the other end of the spectrum, no recordable NAP together with a caliber shift of the nerve (suggesting a neuroma in continuity) may facilitate the surgeon's decision for a grafting procedure without a time consuming internal neurolysis. But there is also evidence from our data that not every nerve lesion in continuity without a NAP needs to be grafted. PMID- 9402586 TI - Cerebral arteriovenous malformation feeding artery aneurysms: a theoretical model of intravascular pressure changes after treatment. AB - OBJECTIVE: A quantitative model may be used to estimate the magnitude of expected pressure changes along the vascular tree with shunt ablation and may provide information to assess the hemodynamic risk of arteriovenous malformation (AVM) treatment. METHODS: A computer model of the cerebral circulation was applied to estimate the changes in intravascular pressure, velocity, biomechanical stress, and shear stress that might be expected from either endovascular or surgical ablation of AVMs. Two AVM sizes and two feeding artery constellations were simulated. The effect of different shunt flows on vascular pressure was modeled. In each simulation, AVMs were occluded in a stepwise fashion. The effects of systemic hypertension and hypotension in various vascular zones were also simulated. RESULTS: As large (1000 ml/min) AVMs were occluded, the mean feeding arterial pressure increased from 18 to 68 mm Hg; the percent-occlusion at half maximal pressure increase was 92%. For medium (500 ml/min) AVMs, feeding arterial pressure increased from 37 to 66 mm Hg; the percent-occlusion at half-maximal pressure increase was 71%. During manipulation of systemic pressure, hemodynamic changes in the circulation close to the nidus were proportionally less than changes in systemic pressure; the degree of proportionality depended on the magnitude of AVM shunt flow. CONCLUSION: In this simulation, shunt obliteration increased pressure in the nidus and feeding arteries with little effect on the proximal circulation. The shunt provided a "buffering" effect, i.e., higher flow fistulas were exposed to smaller variations in intravascular pressure in feeding artery and nidal pressures during manipulation of systemic pressure. PMID- 9402587 TI - Progesterone receptor gene expression in craniopharyngiomas and evidence for biological activity. AB - OBJECTIVE: Previous studies have demonstrated the presence of estrogen receptors in human craniopharyngiomas, raising the possibility that these lesions can be influenced by steroids. To complement these earlier findings, we examined for the presence of progesterone receptor (PR) messenger RNA in surgically removed craniopharyngiomas and performed some studies to determine whether progestogens can exert biological effects on these tumors in vitro. METHODS: Total RNA was extracted from fresh surgically removed craniopharyngiomas and reverse transcribed into cDNA. The polymerase chain reaction was applied to this craniopharyngioma-derived cDNA using amplimers complementary to exons 4 and 7 of the PR gene. Additionally, craniopharyngioma cell cultures were established, and the in vitro effects of progesterone and 6 alpha-methyl-17 alpha hydroxyprogesterone acetate on [3H]thymidine uptake and 17 beta-estradiol oxidoreductase activity were determined. RESULTS: Reversed-transcribed polymerase chain reaction of craniopharyngioma-derived RNA yielded bands of predicted size (389 base pairs) in six of seven tumors studied. Hinfl digestion and direct sequencing of the bands confirmed that the polymerase chain reaction DNA was representative of PR messenger RNA. Treatment of craniopharyngioma cell cultures with progesterone resulted in reduced [3H]thymidine uptake. Both progesterone and 6 alpha-methyl-17 alpha-hydroxyprogesterone acetate powerfully increased oxidative 17 beta-estradiol oxidoreductase activity. CONCLUSION: These results provide evidence that PR messenger RNA can be produced by at least some human craniopharyngiomas and indirectly show that this is translated into biologically active receptor protein. PMID- 9402588 TI - Characterization of a spontaneous murine astrocytoma and abrogation of its tumorigenicity by cytokine secretion. AB - OBJECTIVE: The promise of immunotherapies developed against brain tumors in animal models has not been realized in human clinical trials. This may be because of the routine use of rodent tumors artificially induced by chemicals or viruses that do not accurately portray the intrinsic qualities of spontaneously arising human tumors and that often fail to incorporate the role of immunosuppressants, such as transforming growth factor-beta, that are secreted by human gliomas. From an astrocytoma that arose spontaneously in inbred VM/Dk mice, we have characterized a highly tumorigenic spontaneous murine astrocytoma cell line (SMA 560) that retains features of glial differentiation and naturally produces high levels of biologically active transforming growth factor-beta. We have used this model to determine whether cytokine production by tumor cells will inhibit intracerebral astrocytoma growth. METHODS: Packaging cell lines producing replication-incompetent retroviral vectors were used to transfect the SMA-560 cell line in vitro with the genes encoding the murine cytokines interleukin (IL) 2, IL-3, IL-4, IL-6, tumor necrosis factor-alpha, gamma-interferon, or granulocyte-macrophage colony-stimulating factor or the costimulatory molecule B7.1 (CD80). RESULTS: Mice challenged intracerebrally with 5000 untransfected SMA 560 cells all succumbed to tumor within 30 days, with a median survival of 25 days. In contrast, mice challenged with SMA-560 cells producing IL-2, IL-4, or tumor necrosis factor-alpha each had a more than 400% increase in median survival (P < 0.0001). In these groups, 78.3% (18 of 23 mice), 66.7% (10 of 15 mice), and 60% (6 of 10 mice) of the mice, respectively, remained alive without evidence of tumor for longer than 100 days after the initial tumor challenge. All other cytokines tested and the expression of B7.1 failed to result in an increase in median survival. CONCLUSION: Using a spontaneous astrocytoma model in an inbred mouse strain, we have shown that cytokine production by glial tumors can abrogate their tumorigenicity in vivo despite production of transforming growth factor beta. These results predict that approaches directed at cytokine production within intracerebral astrocytomas may be efficacious in human trials and that the "immunological privilege" of the brain may not be absolute under such conditions. PMID- 9402589 TI - Effect of implant dose/volume and surgical resection on survival in a rat glioma brachytherapy model: implications for brain tumor therapy. AB - OBJECTIVE: This study sought to investigate the effects of implant dose/volume and surgical resection on survival in a rat glioma brachytherapy model. Two doses were investigated to determine a suitable therapeutic range. METHODS: We performed two experiments. Three treatment groups and one control group of male F 344 rats bearing 9L brain tumors 12 days after tumor inoculation were used in the first experiment. Day 12 tumors were an average of 4 to 6 mm in diameter. Animals treated with brachytherapy received a tumor dose of 80 Gy delivered to a 5.5-mm radius volume. Total macroscopic tumor removal was achieved by microsurgical techniques. A subsequent experiment compared the survival of tumor-burdened rats treated with an implant dose of 60 Gy delivered to a 5.5-mm-radius volume with a control group. RESULTS: Surgery alone produced an increased life span of 28.6% over control animals treated with sham surgery and dummy seed implants, a statistically significant increase in survival (P = 0.0023, log-rank test). Brachytherapy alone produced the most significant increase in survival over control animals (P = 0.0001, log-rank test; median survival not attained with an implant dose of 80 Gy delivered to a 5.5-mm-radius volume; and P = 0.0001, increased life span 121% with an implant dose of 60 Gy delivered to a 5.5-mm radius volume). This was not improved by the addition of surgical tumor removal. CONCLUSION: We have demonstrated a relationship between implant dose/volume and survival of tumor-burdened rats in this model that is not improved by the addition of tumor removal. Implications for brain tumor brachytherapy are discussed. PMID- 9402590 TI - Systemic administration of the iron chelator deferiprone attenuates subarachnoid hemorrhage-induced cerebral vasospasm in the rabbit. AB - OBJECTIVE: Iron catalyzed generation of injurious free radicals has been implicated in the pathogenesis of cerebral vasospasm after subarachnoid hemorrhage (SAH). The present study assessed the effects of the iron chelator deferiprone on cerebral vasospasm in an in vivo rabbit model of SAH. METHODS: Twenty-four rabbits were assigned to three groups as follows: SAH plus placebo (n = 8), SAH plus deferiprone (n = 8), or control plus placebo (n = 8). Deferiprone was administered to an additional group of three rabbits that were not subjected to SAH. Drug administration was initiated 8 hours after SAH was induced and was repeated at 8-hour intervals. The animals were killed using perfusion-fixation 48 hours after SAH. Cross-sectional areas of basilar artery histological sections were measured by an investigator blinded to the treatment groups. RESULTS: In placebo-treated animals, the average luminal cross-sectional area of the basilar artery was reduced by 54% after SAH compared to controls (i.e., from 0.272 to 0.125 mm2). The vasospastic response after SAH was attenuated significantly in animals treated with deferiprone (0.208 mm2, representing a 24% reduction). CONCLUSION: Previous experimental studies suggested that iron chelation can be effective in attenuating cerebral vasospasm after SAH. Deferiprone is a recently developed iron chelator that has been extensively evaluated for the treatment of patients requiring chronic blood transfusions. The present study demonstrates that deferiprone is effective in attenuating experimental cerebral vasospasm. Because of its stability, lipophilicity, and ability to penetrate the blood-brain barrier, deferiprone represents an attractive candidate for the treatment of cerebral vasospasm. PMID- 9402591 TI - Bow Hunter's stroke caused by a nondominant vertebral artery occlusion: case report. AB - OBJECTIVE AND IMPORTANCE: Bow hunter's stroke is a consequence of vertebrobasilar insufficiency as a result of mechanical occlusion or stenosis of the vertebral artery at the C1-C2 level by head rotation. In most cases, a dominant vertebral artery is involved. No case of bow hunter's stroke as a result of mechanical occlusion of a nondominant vertebral artery has ever been reported. CLINICAL PRESENTATION: We describe a rare case of Wallenberg's syndrome caused by occlusion of a nondominant vertebral artery induced by head rotation. The patient complained of vertigo and paresthesia of the left face and the right extremities when he rotated his head 45 degrees or more to the right. INTERVENTION: Dynamic angiography revealed that the left vertebral artery was smaller than the right, terminated in a branch of the posteroinferior cerebellar artery, and was stretched and completely occluded at the C1-C2 level with the head rotated 45 degrees to the right. The right vertebral artery was normal when the head was rotated to either the right or the left. Three-dimensional enhanced computed tomography with the head rotated 45 degrees to the right revealed that the left vertebral artery was stretched and occluded by dislodgment between C1 and C2. Cerebral blood flow scintigraphy with head rotation demonstrated that blood flow was decreased in the lower portion of the left cerebellar hemisphere. C1-C2 posterior fixation was performed to prevent life-threatening neurological accidents. CONCLUSION: We emphasize that the diagnosis of bow hunter's stroke should be based not only on angiographic findings but also on hemodynamic studies with head rotation. PMID- 9402592 TI - Stroke related to a dermoid cyst: case report. AB - OBJECTIVE AND IMPORTANCE: We report the case of a woman presenting with sudden neurological deficit, revealing a parasellar dermoid cyst. To our knowledge, this clinicopathological finding is the first reported in the literature. CLINICAL PRESENTATION: A neurological examination of the patient revealed a left hemiparesis, including central facial palsy, which hampered her speech. The well documented neuroradiological work-up (including computed tomography, magnetic resonance imaging, and magnetic resonance angiography) demonstrated right frontorolandic ischemia caused by a right supra- and parasellar dermoid cyst leading to middle and anterior cerebral arterial stenoses. INTERVENTION: Surgical intervention, using a right subfrontopterional approach, was successful. Complete dermoid cyst removal was achieved. The mechanism of the arterial stenoses is extensively discussed and is thought to result from an inflammatory reaction of the basal vessels. CONCLUSION: The patient recovered fully. Nevertheless, postoperative magnetic resonance imaging confirmed cerebral infarction. PMID- 9402593 TI - Traumatic basilar aneurysm after endoscopic third ventriculostomy: case report. AB - OBJECTIVE AND IMPORTANCE: This case illustrates that although endoscopic third ventriculostomy for patients with aqueductal stenosis is successful and minimally invasive, it can have severe, life-threatening complications. CLINICAL PRESENTATION: A 3-year-old girl presented with hydrocephalus and aqueductal stenosis. She underwent endoscopic third ventriculostomy with laser fenestration of the third ventricular floor. During the procedure, she developed a severe intraventricular hemorrhage that required prolonged external ventricular drainage and ultimately ventriculoperitoneal shunting. Despite having a negative angiogram after the procedure, she presented 1 month later with a subarachnoid hemorrhage and a traumatic basilar tip aneurysm. INTERVENTION: The patient underwent a right subtemporal approach with clip ligation of the aneurysm and subsequently had a good recovery. CONCLUSION: Hemorrhagic complications after endoscopic third ventriculostomy are rare. The formation of a traumatic basilar tip aneurysm after this procedure has not been reported in the literature. Laser fenestration of the third ventricular floor may increase the risk of this event. PMID- 9402594 TI - A diffuse white matter ischemic lesion appearing 7 years after stereotactic radiosurgery for cerebral arteriovenous malformations: case report. AB - OBJECTIVE AND IMPORTANCE: Little information is available about radiation-induced complications occurring more than 5 years after radiosurgical treatment for arteriovenous malformations. CLINICAL PRESENTATION: We present a patient with arteriovenous malformations who experienced hemimotor weakness caused by a diffuse white matter necrotic lesion developing 7 years after gamma knife radiosurgery. The original nidus had been too large (24.1 cm3) to be totally covered and irradiated with a peripheral dose of 20 to 25 Gy. Therefore, the lower half of the nidus, which was adjacent to the major feeding artery, had been partially covered with a 30% isodose volume using two target points with an 18-mm collimator. A central dose of 70 Gy was used to obtain 21 Gy at the periphery. Complete nidus obliteration was angiographically confirmed 38 months after radiosurgery. After a 6-year uncomplicated period, this patient experienced a convulsive seizure and then mild right hemiparesis. INTERVENTION: Computed tomography demonstrated a diffuse hypodense area in the left white matter, which had not been revealed by the previous examination. With steroid treatment, this patient achieved clinical improvement, although there was no significant improvement in the computed tomography-demonstrated white matter lesion. CONCLUSION: Although the evaluation of this patient may not be sufficient and further examinations may be necessary, we tentatively conclude that the computed tomography-demonstrated hypodense lesion in this patient is a radiation-related necrotic lesion. Long-term follow-up is crucial, even after the "treatment goal" has been achieved. PMID- 9402595 TI - Ganglioglioma of the spinal cord: report of two rare cases and review of the literature. AB - OBJECTIVE AND IMPORTANCE: The goal of this article is to present the clinical and histopathological features of two rare cases of ganglioglioma occurring in the cervicothoracic and thoracolumbar spinal cord. CLINICAL PRESENTATION: A 4-year old female patient presented with tetraparesis, whereas a 54-year-old woman showed paraparesis of both feet. INTERVENTION: Both tumors could be removed totally by microsurgical techniques. Light microscopically, the tumors in both cases showed basically identical histological features and were diagnosed as benign gangliogliomas. Postoperatively, the two patients did not show improvement. Tumor recurrence was not noted at follow-up examinations within 11 and 24 months after surgery, respectively. CONCLUSION: Ganglioglioma must be considered in the differential diagnosis of tumors affecting the spinal cord. In cases of suspected spinal ganglioglioma showing no sharp delineation from the surrounding tissue, a subtotal tumor removal should be considered to prevent severe neurological deficits. PMID- 9402596 TI - Primary intramedullary primitive neuroectodermal tumor of the spinal cord: case report and review of the literature. AB - OBJECTIVE AND IMPORTANCE: Primary intraspinal primitive neuroectodermal tumors (PNETs) are rare. We report a case and review the literature. CLINICAL PRESENTATION: A 22-year-old woman presented with rapidly progressive paraparesis and neurogenic bladder. INTERVENTION: Preoperative computed tomography myelograms revealed a complete block at T12-L1, consistent with an intramedullary lesion. An urgent operation was performed with gross total tumor removal. The pathological findings were consistent with a PNET. Recurrence was noted within 10 weeks of surgery and has been somewhat responsive to chemotherapy and radiotherapy thus far. A review of the English literature shows that only 13 cases of primary intraspinal PNETs have been reported to date, and the present case is the second one in which the tumor was purely intramedullary. Most of the reported patients survived less than 2 years. CONCLUSION: Primary intraspinal PNETs are rare tumors and carry a poor prognosis. PMID- 9402597 TI - Anterolateral lumbar lipomyelomeningocele: case report and review of the literature. AB - OBJECTIVE AND IMPORTANCE: Meningoceles associated with defects of the abdominal wall are exceedingly rare. One such complex case is presented along with a review of the relevant literature. The current pathophysiological theories and surgical management are discussed. CLINICAL PRESENTATION: A case of a patient with an anterolateral lumbar lipomyelomeningocele associated with multiple congenital anomalies, including defects in the abdominal wall and urogenital system, is presented. The lipomyelomeningocele presented as an expanding abdominal mass. INTERVENTION: Ventriculoatrial shunting and two operations to repair the myelomeningocele were performed to control the expanding abdominal mass. CONCLUSION: This report illustrates that the surgical management of complex lipomyelomeningoceles is similar to the more common types. It also demonstrates that in the infant in whom the intra-abdominal cerebrospinal fluid collection is the primary cause of the symptoms, cerebrospinal fluid shunting may be used to delay the definitive repair until the dura strengthens, thus avoiding the complications of complex repairs of dura with low tensile strength. PMID- 9402598 TI - Telemetric intraventricular pressure measurements after third ventriculocisternostomy in a patient with noncommunicating hydrocephalus. AB - OBJECTIVE: To examine and document intraventricular pressure (IVP) dynamics in an adult after endoscopic third ventriculocisternostomy performed as treatment for hydrocephalus associated with aqueductal stenosis. METHODS: A 30-year-old man who had undergone ventriculoperitoneal shunting at age 21 years for aqueductal stenosis caused by a tectal mass presented with symptoms and imaging studies consistent with shunt malfunction. He underwent urgent ventriculoscopic third ventricular ventriculocisternostomy, which resolved his symptomatology. The existing shunt was not revised. At the time of surgery, a catheter connected to an intracranial pressure TeleSensor device (Radionics, Burlington, MA) was inserted into the ventricular system. Postoperatively, the patient's recovery was assessed by IVP recordings. This system allowed us to record IVP in an awake patient with a functioning third ventriculocisternostomy. RESULTS: We observed an initial postoperative IVP of 17 cm H2O in the supine position, which decreased to 0 cm H2O at 90 degrees of head elevation. The IVP decreased during the first 48 hours postoperatively to 0 to 2 cm H2O when supine. By 1 week postoperatively, the patient's IVP had returned to a baseline of 15 to 17 cm H2O when supine, with a gradual decrease to 0 cm H2O at 30 degrees of head elevation. Three months postoperatively, the patient's IVP in the supine position was 8 cm H2O, with IVP decreasing to 0 cm H2O at 45 degrees of head elevation. Magnetic resonance (MR) imaging performed at that time revealed evidence of flow through the third ventriculocisternostomy. CONCLUSION: We conclude that after an initial period of adjustment, the IVP in this patient returned to an unremarkable baseline despite the novel fluid pathway into the prepontine cistern. This may represent maturation of the breach through the third ventricular floor or brain recovery from a period of high pressure. Also, the shape of the postural IVP curve closely resembled that observed in patients who are not hydrocephalic. These data represent the first documentation of the intraventricular pressure response to ventriculocisternostomy and suggest possible intracerebral responses to this alteration in cerebrospinal fluid flow. PMID- 9402599 TI - Technical accuracy of a neuronavigation system measured with a high-precision mechanical micromanipulator. AB - OBJECTIVE: This study was designed to determine and evaluate the different system inherent sources of erroneous target localization of a light-emitting diode (LED) based neuronavigation system (StealthStation, Stealth Technologies, Boulder, CO). METHODS: The localization accuracy was estimated by applying a high-precision mechanical micromanipulator to move and exactly locate (+/- 0.1 micron) the pointer at multiple positions in the physical three-dimensional space. The localization error was evaluated by calculating the spatial distance between the (known) LED positions and the LED coordinates measured by the neuronavigator. The results are based on a study of approximately 280,000 independent coordinate measurements. RESULTS: The maximum localization error detected was 0.55 +/- 0.29 mm, with the z direction (distance to the camera array) being the most erroneous coordinate. Minimum localization error was found at a distance of 1400 mm from the central camera (optimal measurement position). Additional error due to 1) mechanical vibrations of the camera tripod (+/- 0.15 mm) and the reference frame (+/- 0.08 mm) and 2) extrapolation of the pointer tip position from the LED coordinates of at least +/- 0.12 mm were detected, leading to a total technical error of 0.55 +/- 0.64 mm. CONCLUSIONS: Based on this technical accuracy analysis, a set of handling recommendations is proposed, leading to an improved localization accuracy. The localization error could be reduced by 0.3 +/- 0.15 mm by correct camera positioning (1400 mm distance) plus 0.15 mm by vibration eliminating fixation of the camera. Correct handling of the probe during the operation may improve the accuracy by up to 0.1 mm. PMID- 9402600 TI - Exploration of the solar system and the introduction of advanced technology to neurosurgery: pegasus or pandemonium. PMID- 9402601 TI - Solar system exploration, technology development, and neurosurgery. PMID- 9402602 TI - Charles S. Sherrington (1857-1952) PMID- 9402603 TI - Bovine pericardium for dural grafts: clinical results in 35 patients. PMID- 9402604 TI - Predictability of extracranial/intracranial bypass function: a retrospective study of patients with occlusive cerebrovascular disease. PMID- 9402605 TI - Frameless stereotactic guidance for surgery of the upper cervical spine. PMID- 9402606 TI - Hemianopic visual field defects in children with intracranial shunts: report of two cases. PMID- 9402607 TI - Circadian variation in skin blood flow responses to passive heat stress. AB - To examine whether there is a circadian variation in skin blood flow response to passive heat stress and maximal skin blood flow, which was measured by local warming to 42 degrees C for 45 min, we studied six men at an ambient temperature of 28 degrees C at four different times of day [0400-0700 (morning), 1000-1300 (daytime), 1600-1900 (evening), and 2200-0100 hours (night)], each time of day being examined on separate days. Heat stress at rest was performed by immersing the legs below the knee in hot water (42 degrees C) for 60 min. The esophageal temperature (Tes) at rest was significantly higher in the evening than in the morning. The maximal skin blood flow (SkBFmax) on both sites, back and forearm, did not show a significant difference among the four times of day. The variation in Tes thresholds for cutaneous vasodilation to heat stress was similar to the circadian rhythm in resting Tes. The relationship of the percentage of SkBFmax (%SkBF) with Tes was significantly lower in the morning than in the evening. The results suggest that the maximal skin blood flow during local warming does not show variation over the day, but the sensitivity of vasodilation to passive heat stress shows a circadian variation. PMID- 9402608 TI - Flavor preferences conditioned by intragastric polycose in rats: more concentrated polycose is not always more reinforcing. AB - Prior studies have obtained conditioned preferences for flavors paired with intragastric (IG) infusions of Polycose (hydrolyzed starch) at concentrations of 1-32% over a different flavor paired with IG water. The present study determined if rats would also learn to prefer a flavor paired with concentrated Polycose infusion over a flavor paired with a more dilute Polycose infusion. In Experiment 1, adult female rats were food-deprived and trained during alternating one-bottle sessions (30 min/day) to associate one flavored solution (the CS + 8) with IG infusions of 8% Polycose, a second flavored solution (the CS + 16) with IG infusions of 16% Polycose, and a third flavored solution (the CS-) with IG water infusions. In subsequent choice tests, the rats displayed similar preferences for the CS + 8 and CS + 16 over the CS-, but preferred the CS + 16 to CS + 8 in a direct choice test. A similar preference pattern was obtained in 22 h/day tests with the rats nondeprived. In Experiment 2, new rats were similarly trained and tested but with CS + 16 and CS + 32 solutions paired with 16% and 32% Polycose infusions, respectively. The rats preferred both CS+ solutions over the CS- solution in the short- and long-term tests. However, the CS + 16 was preferred over the CS + 32 by the food-deprived rats in the short-term tests. The two CS+ solutions were equally preferred in the long-term tests with food ad lib. These and other findings indicate that the postingestive reinforcing action of Polycose increases as concentration increases from 1% to 16% but does not increase further, and may actually decrease, at a 32% concentration. The rapid satiating effect of concentrated carbohydrate solutions may limit their reinforcing consequences. PMID- 9402609 TI - Bombesin affects the central nervous system to produce sodium intake inhibition in rats. AB - Bombesin (BN) elicits in the rat important behavioural modifications, including inhibition of food and of water intake. Recently, it has been observed that the peptide also inhibits the intake of sodium chloride. To state whether BN possesses a selective antinatriorexic effect or it elicits only an aspecific depression of ingestive behaviour, we studied the effects of this peptide on the intake of sodium, water or sucrose of Wistar rats after injections into the fourth brain ventricle or into selected brain areas involved in the control of sodium intake, containing BN-like peptides and/or their precursors or specific receptors. We observed that: a) BN (100-200 ng/rat) injected into the fourth brain ventricle inhibits not only the intake of 2% NaCl of sodium depleted rats but also that of water and of 5% sucrose; b) BN (5-50 ng/rat) administered into the nucleus of the solitary tract and the medial amygdala does not influence the intake of these fluids and c) BN (5-50 ng/rat) injected into the paraventricular nucleus does not influence the intake of water and 5% sucrose but potently inhibits that of 2% NaCl. We concluded that the inhibitory effect elicited on salt intake by intracranial administration of BN is selective for this behaviour and is not the expression of an aspecific depression of ingestive behaviour. PMID- 9402610 TI - Effects of interleukin-1 on sexual attractivity in a model of sickness behavior. AB - Interleukin-1 (IL-1), a cytokine secreted by activated macrophages, inhibits sexual behavior in female but not male rats. The present study examined the effects of IL-1 on sexual attractiveness of the injected animal and on the sexual responses exhibited by its mating partner. In Experiment 1, a male rat was placed with an estrous female, injected with either IL-1 beta (2 or 10 micrograms/kg) or saline. Males exhibited more mounts and intromissions per ejaculation and longer ejaculation latencies with IL-1- than with saline-injected females. In a second experiment, a male was placed with two estrous females, one injected with IL-1 beta (5 micrograms/kg) and the other with saline. Males performed less sexual behavior and spent less time with the IL-1-injected female. In a third experiment, an estrous female was placed with two males, one injected with IL-1 beta (5 or 20 micrograms/kg) and the other with saline. IL-1 had no effect on the time spent by the female with each male, and only the high dose reduced proceptive (courtship) behavior. In conclusion, IL-1 administration to females reduces the quality of the sexual act, thus reducing the chances for conception during infection, which is associated with spontaneous abortion and abnormal development of the fetus. In males, the chances for reproduction are less affected by IL-1, possibly because reproduction during infection is not as risky in males as in females. PMID- 9402611 TI - Decreased glucose utilisation does not increase food intake in the marsupial Sminthopsis crassicaudata. AB - The marsupial Sminthopsis crassicaudata increases food intake following a fast. However, the role of metabolic fuel availability, in particular glucose, in the regulation of food intake in this animal is unknown. In this study, we have demonstrated that neither insulin-induced hypoglycaemia nor metabolic blockade of glucose utilisation with 2-deoxy-D-glucose effects food intake compared to saline treated controls, suggesting that mechanisms other than glucose availability are important in the regulation of food intake in this marsupial. These data are discussed in the context of the role of glucoprivation in feeding in other mammals. PMID- 9402612 TI - Caffeine withdrawal symptoms following brief caffeine deprivation. AB - The effects of short-term caffeine deprivation on mood, withdrawal symptoms and psychomotor performance were studied in habitual coffee drinkers. Thirty-one male and female coffee drinkers were tested twice at midday (1130 to 1330 h) 4 h after double-blind administration of 250 mg of caffeine or placebo. Mood and withdrawal symptoms reports were collected by questionnaires. Psychomotor performance was tested with a brief computerized test battery, and causal blood pressure was measured. Caffeine deprivation was associated with decreased vigor and increased fatigue and with symptoms including sleepiness and yawning. Blood pressure was lower by 5-6 mm Hg. No changes in psychomotor performance were observed. Even short periods of caffeine deprivation, equivalent in length to missing regular morning coffee, can produce noticeable unpleasant caffeine withdrawal symptoms in habitual coffee drinkers. Such symptoms may be common side effects of habitual caffeine consumption that contribute to the maintenance of this behavior. PMID- 9402613 TI - Paternal care reduces maternal hyperthermia in Djungarian hamsters (Phodopus campbelli). AB - Throughout lactation, maternal body temperature, nest attendance, activity level and reproductive success of solitary female Djungarian hamsters housed at the recommended ambient temperature of 23 degrees C (Canadian Council on Animal Care guidelines) were compared with those of paired females housed at the same temperature and with solitary females housed at the natural burrow temperature of 18 degrees C. As expected, cooler ambient temperature improved pup survival and weaning weight. Likewise, paternal presence largely compensated for the poor pup growth typical at 23 degrees C. However, the mechanisms were not the same. Females at reduced ambient temperatures were as hyperthermic as females at the higher temperature and spent the same proportion of their day at very high body temperatures. However, the steeper temperature gradient available for passive cooling allowed those females to enhance maternal care by shortening their nest bout absences. In contrast, body temperatures of paired females were tightly regulated compared to the hyperthermia of solitary females and rarely included the highest body temperatures. This alleviation of maternal hyperthermia was not achieved through a reduction in nest attendance. Therefore, maternal hyperthermia in Djungarian hamsters is not essential and may be considered a substantial cost to females when males are not present. PMID- 9402614 TI - Effects of paraventricular lesions on sex behavior and seminal emission in male rats. AB - Oxytocinergic neurons of the paraventricular nucleus (PVN) of the hypothalamus have been implicated in modulating male sexual responses in rats. Previous investigators have shown that cerebrospinal fluid concentrations of oxytocin (OT) increased after ejaculation and that intraventricular administration of OT and electrolytic lesions of the PVN increased temporal measures of male sexual behavior. Recently, we have demonstrated that OT-immunoreactive neurons in the parvocellular subnuclei of the PVN project to lower levels of spinal cord. In the present study, N-methyl-D-aspartic acid lesions, which have been shown to destroy parvocellular PVN neurons while leaving magnocellular neurons intact, were used to evaluate the role of parvocellular neurons on male copulatory behavior and seminal emissions. OT-immunoreactive fibers were reduced in the lower lumbar spinal cord (L5-L6) following N-methyl-D-aspartic acid lesions in the PVN. This reduction was associated with a significant decrease in seminal emission at the time of ejaculation, but mount, intromission and ejaculatory latencies were unaffected. PMID- 9402615 TI - A regional variation of acetylcholine-induced relaxation in different segments of rat aorta. AB - The tension of the isolated thoracic aorta of the rat was measured isometrically to determine if there is a regional variation of the relaxation to acetylcholine (ACh). Endothelium-derived nitric oxide (NO)-dependent relaxation to ACh in the thoracic aorta precontracted with norepinephrine was significantly greater in the middle and distal segments than in the proximal segments. The relaxation to ACh was inhibited by NG-nitro-L-arginine methyl ester, an inhibitor of NO synthase. The relaxation to the NO donor sodium nitroprusside did not differ between the same three segments of the thoracic aorta. These results suggest that there are regional variations in the ACh-induced release of endothelium-derived NO in the rat thoracic aorta. PMID- 9402616 TI - Metabolic fuel privation in hibernating and awake ground squirrels. AB - The nature of metabolic fuel utilization during hibernation and periodic arousal is not completely understood. 2-Deoxy-D-glucose (2DG) and mercaptoacetate (MA) were administered to hibernating ground squirrels. These drugs disrupt glucose and fatty acid oxidation, respectively. Telemetrically recorded body temperature (Tb) was analyzed to determine rate of rewarming from hibernation, duration of euthermia during periodic arousal, and proportion of animals arousing after treatments. 2DG given during hibernation significantly increased latency to regain euthermia, especially during the initial phase of rewarming (from first Tb > 10 degrees C to first Tb > 15 degrees C), without affecting the duration or other features of the ensuing euthermic period; MA did not affect rate of rewarming. MA treatment during hibernation affected thermoregulation after the animals aroused, including an increased duration of euthermia and maintenance of erratic patterns of Tb. The percentage of animals that aroused from hibernation was increased in a dose-dependent fashion by each drug. 2DG and MA treatments had little or no impact on nonhibernating ground squirrels in the cold. We suggest that glucose oxidation is important for rewarming from deep torpor; limited glucose availability cannot, however, support normal levels of euthermia when fatty acid oxidation is compromised. On the other hand, fatty acid oxidation may be less necessary for normal arousal from torpor, but critical for the maintenance of euthermia during the arousal phase. PMID- 9402617 TI - Rat strain differences suggest a role for corticotropin-releasing hormone in modulating sleep. AB - Corticotropin-releasing hormone (CRH) mediates many of the hormonal, behavioral, and autonomic responses to a variety of stressors. There is also evidence suggesting that CRH may be involved in the modulation of physiologic waking. Lewis (LEW) rats possess a hypothalamic gene defect that results in reduced synthesis and secretion of CRH relative to genetically related Fischer 344 (F344) and Sprague-Dawley (Sp-D) rat strains. We therefore hypothesized that LEW rats would spend less time awake, and more time asleep, than either F344 or Sp-D rats. Adult male LEW, F344, and Sp-D rats were surgically provided with electroencephalograph (EEG) recording electrodes, and a thermistor to measure cortical brain temperature [T(cort)]. Additional rats were also provided with a chronic guide cannula directed into a lateral cerebral ventricle. Spontaneous sleep-wake behavior was determined from 48-h recordings of the EEG, T(cort), and body movements from freely behaving, undisturbed rats. Analyses of 48-h recordings from undisturbed animals indicate that LEW rats spend less time awake and more time in slow-wave sleep, relative to the other strains tested. Rapid eye movement sleep did not differ consistently between rat strains. LEW and Sp-D rats exhibit the same degree of waking in response to intracerebroventricular administration of CRH, indicating central mechanisms mediating behavioral responses to exogenously administered CRH are intact in LEW rats. These data provide support for the hypothesis that CRH may be a modulator of waking and sleep. PMID- 9402618 TI - Alarm pheromone induces stress analgesia via an opioid system in the honeybee. AB - Changes of the stinging response threshold of Apis mellifera scutellata were measured on foragers fixed on a holder and stimulated with an electric shock as a noxious stimulus. The threshold of responsiveness to the noxious stimulus increased when bees were previously stimulated with isopentyl acetate, which is a main component of the alarm pheromone of the sting chamber. This effect is antagonised by previous injection of naloxone-hydrochloride (Endo Laboratories Inc.). Results suggest that in the honeybee an endogenous opioid system activated by isopentyl acetate is responsible for modulation of perception for nociceptive stimuli. The resulting stress-induced analgesia in the defender bee would reduce its probability of withdrawal thus increasing its efficiency against enemies. PMID- 9402619 TI - Timing of mating, developmental asynchrony and the sex ratio in mice. AB - According to the developmental asynchrony hypothesis, changing the time of mating within the estrous cycle could alter the interval between completion of blastocyst development and uterine responsiveness for implantation. This may then lead to sex ratio skews in animals that exhibit sex-differential blastocyst development, because uterine stage may now benefit either slow (female) or fast (male) developing blastocysts. To test this hypothesis, the responses of two strains of mice to altered mating dynamics were compared. In a strain that exhibits higher male than female blastocyst developmental rates, sex ratios became significantly female-biased when mated late during the estrous cycle as opposed to early mating. However, timing of mating did not affect sex ratios in a strain with synchronous development of male and female preimplantation embryos. Hence, it is concluded that developmental asynchrony between male and female blastocysts on the one hand, and blastocysts and uterus on the other, are indeed responsible for the effect of timing of mating on litter sex ratios in mice. PMID- 9402620 TI - Recovery from activity-stress ulcer by ad lib feeding in rats. AB - In order to investigate the recovery from activity-stress ulcers by ad lib feeding and/or cessation of running, male Wistar rats were exposed to the activity-stress paradigm, and the rats that revealed hypothermia (their rectal temperature fell below 36 degrees C) were sacrificed either immediately or after several 24 h periods of healing. Rats that were sacrificed immediately after the appearance of hypothermia and those that were exposed to restricted feeding plus cessation of running revealed severe activity-stress ulcers, whereas few ulcers were observed in rats given ad lib-feeding and those that were given ad lib feeding plus cessation of running. Although no significant differences in relative weights of spleen and thymus were obtained among the different recovery conditions, the relative weights of the adrenal glands were highest in the restricted feeding plus cessation of running group, whereas, the other animals exposed to the activity-stress paradigm showed no differences. These results indicate that activity-stress ulcers recover under conditions of ad lib-feeding within 24 h, but they are not influenced by cessation of running. These data also suggest that organ weights are not affected by any manipulations employed in the present study. PMID- 9402621 TI - Failure to change exploration or saccharin preference in rats exposed to chronic mild stress. AB - Chronic mild stress (CMS) exposes animals to unpredictable stressors. Reduced consumption of sucrose or saccharin solutions by CMS rats has been used as a putative measure of anhedonia, typical of depression. Our objective was to determine whether saccharin consumption and preference and suppression of exploratory and rearing behaviors in the open field were reliable indicators of CMS-induced behavioral depression. In Experiment 1, male Wistar rats subjected to 6 weeks of CMS consumed significantly less food and gained less weight than controls. CMS did not effect saccharin intake, or preference, measured in a two bottle test with water. CMS rats exposed to a novel open field showed increased exploration and rearing. In a second test, performed immediately after a novel stress of restraint, there were no differences in exploratory or rearing behavior of CMS and control rats. In Experiment 2, CMS was reduced to 3 weeks and rats were single or group housed in their home cages. Open field activity of CMS rats was similar to that in Experiment 1. Saccharin preference of CMS rats was significantly suppressed when tested after 24 hours of water deprivation, but was not different from controls after 5 hours of water deprivation. In the final experiment Sprague Dawley rats behaved the same as Wistar rats in the CMS paradigm. Therefore, the CMS protocol used in these experiments did not induce behaviors indicative of depression but did cause a mild anorexia and weight loss. Saccharin intake of CMS rats was dependent upon their dehydration state and could not be attributed to stress-induced anhedonia. PMID- 9402622 TI - Behavioral specificity of the bitter taste gene Soa. AB - In mice, aversion to the bitter acetylated sugar sucrose octaacetate (SOA) is determined by a single genetic locus with three alleles. SWR/J (SW) inbred mice are SOA tasters: They avoid many compounds characterized as bitter-tasting by humans, at concentrations to which C3HeB/FeJ (C3:SOA demitasters) mice are less sensitive. C3.SW-Soa(a) congenic taster mice contain the taster allele transposed to a 99% C3 bitter-insensitive genetic background. SW, C3, C3.SW-Soa(a) congenic taster, and C3.SW demitaster mice were behaviorally tested with a series of 48-h two-bottle preference tests to determine the influence of the Soa(a) taster allele on sensitivity to a variety of bitter-tasting compounds. Soa allelic variation had a major effect on sensitivity to 0.003-1.0 mM SOA and several concentrations of the bitter-tasting alkaloids brucine, strychnine, and quinine. Effects were also found for 0.1 mM denatonium and 1 mM propylthiouracil. For caffeine, cycloheximide, thiamine, and two nonbitter compounds (NaCl and calcium hydroxide), the SW mice avoided lower concentrations than the other strains, but this avoidance was not due to the Soa(a) allele because both the C3 inbred and C3.SW-Soa(a) congenics were less sensitive. These results suggest the Soa gene product influences sensitivity to a subset of bitter-tasting compounds. PMID- 9402623 TI - Relation between parotid saliva flow and composition and the perception of gustatory and trigeminal stimuli in foods. AB - The objective of this study was to investigate whether and how parotid saliva flow and composition correlated with the perception of gustatory and/or trigeminal stimuli in foods. Thirty (15 male and 15 female) subjects tasted seven foods or beverages (lemonade, beer, wine, soup, methyl cellulose, peanut butter, and crackers) with three levels each of a gustatory or trigeminal stimulus and rated the perceived intensity of the corresponding sensation over time using the time-intensity (TI) method while their parotid saliva was being collected. Salivary flow rates of males were significantly higher than those of females for all stimuli (p < 0.001). That did not translate, however, into consistent differences in perception of sensory attributes between males and females. Significant positive correlations were found between saliva flow and (1) TI parameters for adhesiveness of peanut butter and cohesiveness of mass of crackers (p < 0.05 or lower) and (2) time from intake to swallowing of crackers and peanut butter (p < 0.05). No correlations were found between saliva composition (e.g., sodium and total protein) and TI parameters. These results indicate that parotid saliva flow may correlate with the perception of some texture and mouthfeel attributes (presumably through oral work and bolus formation) but not with that of the taste attributes examined in this study (at the concentrations studied). PMID- 9402624 TI - Modifications of operant thermoregulatory behavior of the young pig by environmental temperature and food availability. AB - Piglets (Sus scrofa domesticus) were weaned at 14 days and acclimated to 10 ("cold animals") or 35 degrees C ("warm animals"). Cold animals were either fed ad lib. (10HH) or maintained on a high (10H) or low (10L) nutrition plane. Warm animals were maintained on a high (35H), low (35L), or a very low (35LL) nutrition plane. After 3 weeks of acclimation, operant thermoregulatory behavior (animals pressed a lever to turn on a heater) at 15 degrees C was assessed immediately following or 22 h after a meal. It was found that cold piglets or those on low nutrition planes produced more heat reinforcements than warm piglets and those on high nutrition planes. All animals tested 22 h after a meal produced more reinforcements than when tested immediately after a meal. Rectal temperature before the sessions of operant heating was lower in cold animals and those on low nutrition planes than in warm animals and those on high nutrition planes, but the difference dissipated after 90 min of access to heat. Piglets were then "deacclimated" for 24 h at 25 degrees C and their behavior observed once again. Deacclimation eliminated the effect of acclimation temperature on body temperature and behavior but it did not eliminate the effect due to nutrition. There was no interaction between temperature of acclimation and nutrition. The experiments demonstrate that the body temperature of the pig is independently affected by environmental temperature, the quantity of food eaten by the animal, and a possibility to use behavior. The behavioral drive is to reach body temperature which can be defined as a thermal set-point (temperature at which there is no need for a thermoregulatory response to occur) for behavioral thermoregulatory responses. Adaptation to different environmental conditions does not affect this behavioral set-point, but it can involve a temporary shift in the set points for specific autonomic thermoregulatory responses. PMID- 9402625 TI - Behavioural lateralisation of the tetrapod type in the zebrafish (Brachydanio rerio). AB - Visual lateralisation resembling that found in a bird (domestic chick) is here demonstrated in a teleost (zebrafish, Brachydanio rerio). Zebrafish predominantly view objects with the body axis close to facing the object (0-20 degrees on either side of facing). Strange objects were viewed at first exposure chiefly with the right frontal field; so was a complex and unfamiliar scene made up of familiar components. In a second trial, using the same stimulus or scene, left frontal viewing tended to be used instead. A familiar partner (a fish of another species) was viewed left frontally. The domestic chick also uses the left eye to view familiar stimuli, shifting to the right when it has to decide what response is appropriate to the object at which it is looking. An empty scene in which nothing could be concealed (and so no response was called for) was viewed by zebrafish with the left eye from the start. In zebrafish and the chick, the right eye is used when it is necessary to inhibit premature response, in order to sustain viewing until a decision is reached, and the left is used when it is necessary to keep an eye on a familiar or clearly empty scene. The findings suggest homology of cerebral lateralisation in teleost fish and tetrapods. PMID- 9402626 TI - D3 dopamine receptor-deficient mouse: evidence for reduced anxiety. AB - Mice without functional D3 dopamine receptors were examined in two animal models for anxiety: the open-field test and the elevated plus-maze test. In the open field, D3 receptor-deficient mice (D3-/-) entered the center significantly more often than normal (D3+/+) littermates, suggesting an anxiolytic-like effect of the D3 receptor mutation. Consistent with this finding, D3-/- mice entered open arms of the plus maze significantly more often and longer than D3+/+ littermates, but did not differ in closed-arm entries, an index of general activity. Heterozygous (D3 +/-) animals showed intermediate behavioral changes. We interpret these results as indicative of reduced anxiety in mice without D3 receptors. Our findings thus suggest that D3 dopamine receptors are involved in the regulation of anxiety. PMID- 9402627 TI - Greater behavioral effects of stress in immature as compared to mature male mice. AB - The effect of sexual maturity on behavioral effects of stress was examined in male mice. Immature (4-week-old) or mature (8-week-old) animals were subjected to either social stress (exposure to an isolated adult male) or restraint stress for 5 days and examined for body weight, food intake, or plus-maze behavior. Social stress reduced food intake, body weight, and open-arm entries in 4-week-old but not 8-week-old mice. Restraint reduced body weight in 4-week-old but not 8-week old mice. It is concluded that immature male mice show greater behavioral disturbances after stress than their mature counterparts. The findings are in agreement with much anecdotal evidence that children are more vulnerable to stress than adults. PMID- 9402628 TI - Evidence that adaptation to cold water swim-induced analgesia is a learned response. AB - Controlling for novelty of the test context, the present experiment determined if adaptation to forced cold water swim stress-induced analgesia is learned through Pavlovian conditioning. Following baseline measurement of pain sensitivity, Group A swam in Context A for 3 min and, 8 h later, sat in Context B for 3 min. The conditions were reversed for Group B. All rats were given a tail-withdrawal test immediately after swimming or sitting in each context. On the first test day, conducted 24 h after the completion of the adaptation phase, all rats swam in Context A for 3 min, and tail-withdrawal latencies were immediately obtained after the swim. On the second test day, 24 h later, all rats swam in Context B, with tail-withdrawal latencies measured immediately thereafter. Both groups showed significantly less analgesia when tested in the adaptation context in which they previously swam than in the other context. These data provide strong evidence that adaptation to stress-induced analgesia is a learned response. PMID- 9402629 TI - The postpubertal change in the playful defense of male rats depends upon neonatal exposure to gonadal hormones. AB - The pattern of playful defense used during play fighting by male rats (Rattus norvegicus) castrated at birth was compared to that of sham-operated and untreated controls during the juvenile phase and after puberty. The neonatal castrates failed to exhibit the age-related changes in playful defense present in intact male rats of the same age. Following puberty, control rats, but not neonatal castrates, switched from juvenile to more adult-typical defensive tactics. That the neonatal castrations were effective was independently shown by the animals' failure to exhibit the asymmetries in weight and play behavior indicative of dominance-subordinance relationships present in normal adult males. A previous study found that castration following weaning did not prevent the pubertal change in playful defense, but did block the formation of dominance subordinance relationships. Therefore, it appears that the age-related shift in playful defense is a feature of the development of play fighting in males that is likely preprogrammed by gonadal hormone exposure in the perinatal period. PMID- 9402630 TI - Measurement of resuspended aerosol in the Chernobyl area. I. Discussion of instrumentation and estimation of measurement uncertainty. AB - Results of measurements of the resuspended radioactive aerosols in the Chernobyl area are presented which were obtained soon after the Chernobyl reactor accident and in a European project in 1992-1993. The measurements were carried out with the intention of obtaining a data base for dose assessment of resuspended radioactive particles. Potential significant dose contributions may result from inhalation and secondary contamination due to resuspended radionuclides. In this first article of a series of three papers, the instrumentation and the measurement uncertainties are discussed. An effort was made to sample quantitatively giant aerosol particles (particles larger than 10 microns aerodynamic diameter) as well. The comparison of the samplers shows, in general, an agreement of concentration measurements of 137Cs and 7Be within a factor of two. One sampler was identified with larger discrepancies and needs additional investigation of its sampling characteristics; for another device, the recalibration of the analysing system is recommended. Ordinary integrating samplers have a loss of about 30% in 137Cs activity compared to an isokinetic sampler collecting giant particles as well. The mean ratio of 137Cs activity concentration between an instrument sampling only particles larger than 10 microns and an ordinary integrating sampler is 0.39 +/- 0.15 during anthropogenic enhanced resuspension. These findings demonstrate the significant contribution of giant particles to resuspended airborne radioactivity. The results of this study concerning integral measurements during wind-driven resuspension proved to be in good agreement with previously published data on resuspension. PMID- 9402631 TI - Exposure levels for persons involved in recovery operations after the Chernobyl accident. Statistical analysis based on the data of the Russian National Medical and Dosimetric Registry (RNMDR). AB - We present a detailed description of dosimetric data entered in the Russian National Medical and Dosimetric Registry (RNMDR) for emergency workers (liquidators) involved in recovery operations (RO) after the Chernobyl accident. The data on the absorbed doses from external exposure are based on the documents given to liquidators by organizations that performed dosimetric monitoring in the zones of operation. Using the data on external doses currently available in the RNMDR for 119,416 liquidators (78.4% of the total number of 152,325 persons), different statistical characteristics were derived to assess the reliability of the information. The paper also discusses dose distributions according to the date of beginning work in the RO zone [up to 250 km from the Chernobyl nuclear power plant (NPP)], on the distance of the settlement where the liquidators were accommodated or worked from the NPP, and on the duration of their stay in the RO zone. To analyse the reliability of the dosimetric data, the notion of an effective exposure dose rate (EEDR), i.e. the ratio of the dose registered in RNMDR and the duration of stay in the RO zone, was introduced for each liquidator, and corresponding statistical characteristics for the distribution of EEDR depending on the date of entry into the RO zone and distance from the place of residence or work to the NPP were obtained. The analysis for different groups of liquidators shows that the dosimetric information of the RNMDR is, as a statistical aggregate, generally consistent with the data on the radiation situation in the RO zones. PMID- 9402632 TI - Cancer risks in the Kaluga oblast of the Russian Federation 10 years after the Chernobyl accident. AB - Cancer morbidity and mortality were studied in areas of the Kaluga oblast contaminated with radionuclides. The main objective of the study was to assess the influence of radiation exposure on existing levels of cancer morbidity and mortality. Time trends and relative population risks were analysed. Based on this analysis, it was concluded that the current levels of morbidity from cancers among the populations residing in the studied areas were primarily a result of a complex of factors which predated the exposure from the Chernobyl accident. However, there seems to be an unfavourable trend concerning malignant neoplasms of the respiratory organs for women residing in the contaminated areas. To date, no statistically significant effect of radiation on cancer morbidity (except for thyroid cancer in women) has been noted. The levels of cancer morbidity and mortality in the contaminated areas generally reflect the changes in cancer incidence in the oblast as a whole. The findings are consistent with international data on latent periods for the induction of radiogenic cancers and the biological effects for similar levels of exposure to populations residing in contaminated territories. Further studies are necessary in order to monitor possible effects that are related to the accident. PMID- 9402633 TI - Dynamics and distribution of radiocaesium in broiler chicken. AB - The distribution and biological half-life of radiocaesium (137Cs) in broiler chickens after three oral applications (in course of 1 day at the age of 14 days) of artificially contaminated feed mixture were studied. There was a rapid uptake of the orally administered 137Cs (within a few hours) and also a rapid loss of 137Cs which varied in the different organs (the initial biological half-life was: liver 0.6 day, intestine 0.6 day, breast meat 2 days, leg meat 1.2 days). More than one-half of the total administered 137Cs activity (55%) was excreted from the body within the 1st day after dosage, and after 14 days more than 90% had been excreted. The highest accumulation of 137Cs occurred in meat (50%-90%), and the proportion of total activity in breast and leg meat varied during decontamination. The transfer of radiocaesium from feed into the chicken body (measured as ratios of the 137Cs activity concentrations in the organ to the 137Cs activity concentration in the applied dose) 1 day after application was: 0.0220, 0.0294, 0.0216 and 0.0195 for breast meat, leg meat, intestine and liver, respectively. Significant differences between the values were demonstrated (P < 0.05) except between those of breast meat and intestine. For the first 3 days there was a higher proportion of 137Cs activity in leg meat, whereas from the 4th day a greater part of total activity was found in breast meat. The latter results were confirmed in a subsequent study. Data from this study suggest that if broiler chickens are contaminated by radiocaesium to a level of 5 kBq/chicken in the course of 1 day at the age of 14 days, then immediate feeding with uncontaminated feed mixture for 18 days should be effective in decontaminating the chicken's meat below the intervention levels for radiocaesium in animal products, i.e. below 1000 Bq. kg-1. PMID- 9402634 TI - Biological dosimetry: the potential use of radiation-induced apoptosis in human T lymphocytes. AB - An assay for biological dosimetry based on the induction of apoptosis in human T lymphocytes is described. Radiation-induced apoptosis was assessed by flow cytometric identification of cells displaying apoptosis-associated DNA condensation. CD4 and CD8 T-lymphocytes were analysed. They were recognized on the basis of their cell-surface antigens. Four parameters were measured for both cell types: cell size, granularity, antigen immunofluorescence and DNA content. Apoptosis was quantified as the fraction of CD4-, or CD8-positive cells with a characteristic reduction of cell size and DNA content. At doses below 1 Gy, levels of radiation-induced apoptosis increased for up to 5 days after irradiation. Optimal dose discrimination was observed 4 days after irradiation, at which time the dose-response curves were linear, with a slope of 8% +/- 0.5% per 0.1 Gy. In controlled, dose-response experiments the lowest dose level at which the radiation-induced apoptosis frequency was still significantly above control was 0.05 Gy. After 5 days post-irradiation incubation, intra- and interdonor variations were measured and found to be similar; thus, apoptotic levels depend more on the dose than on the donor. The results demonstrate the potential of this assay as a biological dosimeter. PMID- 9402635 TI - PARATI--A dynamic model for radiological assessments in urban areas. Part II. Specifications of individuals and populations, their radiation exposures and variabilities. AB - After a large-scale contamination of an urban area with gamma-ray emitting radionuclides (e.g. caesium isotopes) decision makers will need guidance as to its potential radiological consequences and to optimum means of mitigation. To provide such information, a dynamic radioecological model PARATI has been developed and used to simulate the contamination of realistic urban environments in a computer model and to estimate the various radiation fields in such environments. In this study, the computer-simulated realistic behaviour and movements of individuals and populations in such radiation fields are described, and the resulting radiation exposures and their variabilities are estimated. For the scenarios considered, the doses of individuals in the same contaminated environment may vary by more than one order of magnitude. Studies on population habits and on the behaviour of radionuclides on important urban surfaces even a long time after the contamination might reduce the uncertainty considerably. PMID- 9402636 TI - Thyroid cancer incidence in the Ukraine after the Chernobyl accident: comparison with spontaneous incidences. AB - The thyroid cancer incidence in the Ukraine among those born in the period 1968 1986 was analyzed with the aim to identify the enhancement due to the Chernobyl accident. Since any Ukrainian data referring to the time period before the accident are scarce and the variation of spontaneous incidences in other countries is immense, the Ukrainian incidences in the period 1986-1989 were used to estimate the baseline risk. Following 1990, the incidence in the southern part of the Ukraine increased by about 30%, independent of age. In the other parts the increase of the incidence depended on age at exposure. In the age group of 9-year old children, the incidences in three regions defined as the 'high-dose area', the northern, and the middle oblasts, increased by factors of 50, 20, and 6, respectively. These rates (1991-1995) are well above spontaneous rates in other countries. In the age group of 17-year-old juveniles, the incidence increased by a factor of 6 for the 'high dose area' and in the three northern oblasts, whereas in the nine 'middle' oblasts it was similar to the incidence of the 'southern' Ukraine. These rates are within the range found in other countries. PMID- 9402637 TI - The Semipalatinsk nuclear test site: a first assessment of the radiological situation and the test-related radiation doses in the surrounding territories. AB - As a result of atmospheric nuclear tests at the Semipalatinsk test site 'Polygon', adjacent territories were contaminated by radionuclide fallout. The population of some districts in the Semipalatinsk oblast were exposed to elevated levels of radiation. Contamination and exposure mostly resulted from early atmospheric tests. The radiological situation of the Semipalatinsk oblast is described. Effective dose estimates due to external and internal exposure attributable to the 1949 and 1953 tests in villages near the Polygon range from 70 mSv to 4470 mSv. PMID- 9402638 TI - No evidence for increased tumor rates below 200 mSv in the atomic bomb survivors data. AB - We investigated for which doses a significantly increased tumor rate can be seen in the RERF Life Span Study data sets on mortality or incidence of solid tumors. No significant increase was found below about 200 mSv. PMID- 9402639 TI - on: 'No evidence for increased tumor rates below 200 mSv in the atomic bomb survivors' data'. PMID- 9402640 TI - Radiation risk estimates for leukemia and thyroid cancer among Russian emergency workers at Chernobyl. PMID- 9402641 TI - Role of neurotrophins in synapse development and plasticity. AB - Neurotrophic factors are traditionally viewed as secretory proteins that regulate long-term survival and differentiation of neurons. The role of neurotrophic factors in the structural integrity of the nervous system makes them attractive candidates as therapeutic agents for neurodegenerative diseases. However, the fact that expression of many neurotrophic factors in the central nervous system is rapidly enhanced by neuronal activity suggests a new role for these factors in activity-dependent processes, such as synaptic development and plasticity. A series of recent studies has provided strong evidence for this novel function of neurotrophic factors. The neurotrophin family of proteins has been shown to acutely potentiate synaptic transmission at the neuromuscular junction and in the brain. These factors are also involved in the maturation of the neuromuscular synapses and in the development of synapses in the visual system. Gene targeting and physiological experiments demonstrate that brain-derived neurotrophic factor (BDNF) plays an important role in long-term potentiation (LTP), a cellular model for learning and memory. These findings have brought together two hotly pursued areas of neuroscience, namely, the function of neurotrophic factors and the mechanisms for synaptic plasticity. Continuous studies in this new field will help understand how synapses develop and function in the brain, and may have significant implications in treating learning disorders in both children and adults. PMID- 9402642 TI - Neurotrophin regulation of the developing nervous system: analyses of knockout mice. AB - The neurotrophins, NGF, BDNF, NT3 and NT4, are one family in a growing repertoire of neurotrophic factors. The neurotrophins have long been implicated in neuronal survival and recent studies from mice with targeted disruptions of the neurotrophin genes confirm this role, but also reveal that the action of the neurotrophins is more complex, and in some instances more interactive, than originally envisaged. Lack of functional NGF, BDNF and NT3 genes results in severe neuronal deficits and an early postnatal death. However, NT4 is unique among the neurotrophins and while the absence of NT4 does result in limited sensory neuron loss these mice do not die early, suggesting that NT4-dependent neurons are not critical for survival. Phenotypic analyses of mice lacking neurotrophin receptors, TrkA, B and C, confirm that TrkA is the functional receptor for NGF, TrkB acts as the primary receptor for BDNF and NT4, and NT3 signals primarily through TrkC. However, the finding that TrkC mutant mice have a less dramatic phenotype than their NT3 counterparts implicates NT3 in signaling via receptors other than TrkC. Further studies, using combinatorial Trk and neurotrophin deletions, reveal that while BDNF and NT4 subserve distinct neuron populations in most cases, other neuron sub-populations can be supported by either BDNF or NT4, providing evidence for compensatory actions between neurotrophins. As a mechanism to explain programmed cell death that occurs in the developing nervous system, recent studies examining neurotrophin gene-dosage effects suggest that the availability of neurotrophins, NGF, BDNF and NT3, may be limiting for some neuron populations. In addition, the proposed switch in neurotrophin dependency for some neuron populations is now being determined using neurotrophin mutant mice. We discuss these and other recent findings on neurotrophin requirements for the developing nervous system. PMID- 9402643 TI - The neuroimmune hypothesis in Parkinson's disease. AB - The role of immune mechanisms in the pathogenesis of Parkinson's disease is still a matter of controversy. Immunological abnormalities have been reported in various brain areas and in peripheral immune parameters. Beside antigen specific findings which might indicate an autoimmune process directed against dopaminergic neurons, non-antigen specific abnormalities have been found, which could be caused either by a disease specific process and/or by immunological properties of various anti-parkinsonian drugs, e.g. bromocriptine. Immune reactions may imitate or maintain the degenerative process. In this case possible neuroprotective strategies interfering with immune functions could be developed. Further investigations are necessary to elucidate the impact of immune mechanisms on the degenerative process in Parkinson's disease. PMID- 9402644 TI - The neurobiological basis of movement initiation. AB - Simple reaction time (RT) is defined as the elapsed time between presentation of a single stimulus and onset of movement. In choice RT, there are at least two stimuli, requiring two distinct responses. The neurobiological basis of RT in humans has mostly been evaluated in patients with Parkinson's disease or cerebellar disease. Lesion studies in animals have assessed the different contributions of various subregions of the basal ganglia and the cerebellum. There is a prolongation of simple RT and in some cases of choice RT in Parkinson's disease. Both simple and choice RT are susceptible to modulation by brain dopamine levels. However, such is not invariably the case, attesting to the contribution of non-dopaminergic neurons in the sensori-motor slowing found in Parkinson's disease. An increase in simple RT and in choice RT are found in patients with cerebellar atrophy. The initiation of fast ballistic movements is associated with the dentate efferent system. This system is modulated by dopaminergic and glutamatergic pathways to the striatum. PMID- 9402645 TI - The regulation of motor control: an evaluation of the role of dopamine receptors in the substantia nigra. AB - The importance of the nigrostriatal dopaminergic pathway in motor control is widely accepted and it is generally believed that the motor symptoms of Parkinson's disease result solely from reduced release of dopamine from terminals in the striatum. Over recent years there has been a growing body of evidence which suggests that dendritic dopamine release in the substantia nigra is of importance in the regulation of neuronal activity and behaviour. This evidence is reviewed together with a description of our recent findings that show nigral dopamine receptors are essential for the maintenance of normal muscle tone. It is concluded that current views of the basal ganglia circuitry involved in motor control need to be re-evaluated to take into account these recent reports. A scheme is suggested to explain how dopamine mechanisms in the substantia nigra regulate motor activity. PMID- 9402646 TI - Primary motor cortex reorganization in a long-term monkey amputee. AB - The primary motor cortex (M1) was mapped with intracortical microstimulation (ICMS) in a 15 year-old macaque whose right upper extremity was amputated at the shoulder joint prior to 2 years of age. Movements of the right shoulder girdle and stump were evoked by ICMS throughout the left M1 upper extremity region. The size of the left M1 upper extremity region contralateral to the amputated arm was not appreciably different from the size of the right upper extremity region contralateral to the intact arm. Long stimulus trains and/or higher stimulus currents were needed to evoke detectable movements at significantly more loci in the left than in the right M1 upper extremity region. These observations would be consistent with unmasking of a high threshold representation of shoulder musculature that normally exists throughout the central core of the upper extremity region, where it underlies a lower threshold representation of the distal forelimb. Alternatively, invasion of the de-efferented distal forelimb core by surrounding shoulder representation may have occurred. Differences between the limited M1 reorganization observed in the present study and the more extensive reorganization of S1 observed in other studies may reflect fundamental differences between M1 and S1, and/or differences in the extent of de efferentation versus deafferentation. PMID- 9402647 TI - Organization, development, and effects of infraorbital nerve transection on galanin binding sites in the trigeminal brainstem complex. AB - Previous experiments from this laboratory have indicated that transection of the infraorbital nerve (ION, the trigeminal [V] branch that supplies the mystacial vibrissae follicles) at birth and in adulthood has markedly different effects on galanin immunoreactivity in the V brainstem complex. Adult nerve transection increases galanin immunoreactivity in the superficial layers of V subnucleus caudalis (SpC) only, while neonatal nerve transection results in increased galanin expression in vibrissae-related primary afferents throughout the V brainstem complex. The present study describes the distribution of binding sites for this peptide in the mature and developing V ganglion and brainstem complex and determines the effects of neonatal and adult ION damage and the associated changes in galanin levels upon their distribution and density. Galanin binding sites are densely distributed in all V brainstem subnuclei and are particularly dense in V subnucleus interpolaris and the superficial layers of SpC. They are present at birth (P-0) and their distribution is similar to that in adult animals. Transection of the ION in adulthood and examination of brainstem 7 days later indicated marked reductions in the density of galanin binding sites in the V brainstem complex. With the exception of the superficial laminae of SpC, the same reduction in density remained apparent in rats that survived > 45 days after nerve cuts. Transection of the ION on P-0 resulted in no change in the density of galanin binding sites in the brainstem after either 7 or > 60 days survival. These results indicate that densely distributed galanin binding sites are present in the V brainstem complex of both neonatal and adult rats, that they are located in regions not innervated by galanin-positive axons, and that their density is not significantly influenced by large lesion-induced changes in the primary afferent content of their natural ligand. PMID- 9402648 TI - The effects of skin temperature on the detection and discrimination of tactile stimulation. AB - Detection thresholds and intensity-difference thresholds were measured on four subjects ranging in age from 19 to 22 years. The stimuli were 250-Hz bursts of vibration applied through a 3.0 cm2 contactor to the thenar eminence of the right hand. Detection thresholds were substantially higher at 20 degrees C than at 30 degrees or 40 degrees C and were only slightly higher at 40 degrees C than 30 degrees C. When the intensity-difference threshold was expressed in relative terms, as the proportion by which two stimuli must differ in amplitude to be discriminated (delta phi/phi), discrimination capacities were unaffected by surface-skin temperature. The results are consistent with the hypothesis that surface-skin temperature alters the sensitivity of tactile receptors, and that, because of the 'near miss' to Weber's law, the relative difference threshold is unaffected substantially by skin temperature. It was concluded that, at least a partial explanation of the 'near miss' lies in the fact that, at low to moderate sensation levels, the P channel is exclusively activated whereas, at moderate to high sensation levels, because of recruitment of activity in Non-Pacinian channels, neural information for intensity discrimination is additionally provided by channels with superior discriminative capacities. PMID- 9402649 TI - Perceptions of effort and heaviness during fatigue and during the size-weight illusion. AB - Previous work has shown that force perception and the sense of motor effort are different attributes of sensorimotor function. This study explores the hypothesis that one reason force and effort perceptions are distinct is to inform an individual of impaired motor function when muscular force lags effort. This hypothesis predicts that effort and force perceptions will dissociate when motor function is impaired by fatigue but not during the size-weight illusion. All subjects reported a distinct increase in effort when lifting a standard test weight as fatigue developed. When fatigue was sufficiently marked so that they could barely lift the test weight, they rated their effort as similar to that required to lift a maximal weight in the unfatigued state. The perceived heaviness of the test weight also increased as fatigue developed, but this fatigue-weight illusion was smaller than the increase in effort for all subjects and displayed greater variability. In contrast, both the perceived weight of a small object and the effort required to lift it increased in parallel when small and large objects were lifted sequentially. The size-weight and size-effort illusions appear to be examples of a common phenomenon in which perceptual experience is rescaled to maintain acuity under different working conditions. The fatigue-weight illusion also has the effect of increasing perceptual acuity as the subject's weight lifting range decreases due to fatigue. PMID- 9402650 TI - Matching object size by controlling finger span and hand shape. AB - The ability of human subjects to accurately control finger span (distance between thumb and one finger) was studied. The experiments were performed without visual feedback of the hand and were designed to study the dependence of accuracy on object size, shape, distance, orientation and finger configuration. The effects of finger combination and sensory modality used to perceive object size (vision and haptics) were also studied. Subjects were quite proficient at this task; the small errors tended to be predominantly negative, i.e., finger span < object size. The thumb-little finger combination was less accurate than the other finger combinations, irrespective of the sensory modality used. Subjects made larger under-estimating errors when matching the size of cylinders than when matching cubes and parallelepipeds. No effect of viewing distance, object orientation and finger configuration was found. Accuracy in matching object size was not dependent on the sensory modality used. The question of how the individual degrees of freedom of the fingers and thumb contributed to the control of finger span was also addressed. Principal components analysis showed that two components could characterize the hand postures used, irrespective of object size. The amplitude of the first principal component was constant, and the amplitude of the second scaled linearly with object size. This finding suggests that all of the degrees of freedom of the hand are controlled as a unit. This result is discussed in relation to the 'virtual finger' hypothesis for grasping. PMID- 9402651 TI - The neocortical local circuit--a research workshop held in Sde-Boker, Israel, May 4-8, 1997. AB - This report does not capture the raw vitality of the Sde-Boker workshop. We did not anticipate that so many presentations would focus on the importance of spike timing in the operations of cortical microcircuits and the profound effect of this on other cortical and subcortical networks. The roles of individual spikes, temporal firing patterns, neuronal synchronization, intrinsic neuronal and network oscillation, coincidence detection, integration by individual cells or even individual dendritic spines were also strong themes of nearly all presentations. The cortex could be viewed as a network, which, is similar to an orchestra, where each neuron (musician) generates particular timed spike patterns (notes) for which 'every spike counts' (the symphony). This assertion would have been the butt of considerable skepticism less than a decade ago. The conference led to a view that the cerebral cortex is a spatially distributed timing network. As such, the consensus reached provides a conceptual framework for exciting new discoveries in the next decade. One of the truly exciting outcomes of the workshop was the seamless integration of concepts derived from studies employing direct approaches such as brain slices to cell cultures and computer simulations to whole animal physiology. The availability of an ever expanding armamentarium of increasingly sophisticated techniques and cross-fertilization promises develop the field in a direction that is as flexible and as subtle as the cortical tissue it seeks to explain. It was perhaps symbolic that this meeting was held at a place in the desert where David Ben-Gurion, the first Prime Minister of the modern state of Israel, had a vision of pioneers settling the untamed Negev that had intrigued and intimidated mankind for so long. The hauntingly beautiful Negev, like the cerebral cortex, was an unyielding frontier. Now thanks to modern ingenuity and technology the Negev can, like the cerebral cortex, finally be conquered. PMID- 9402652 TI - Structure, function and expression on blood and bone marrow cells of the urokinase-type plasminogen activator receptor, uPAR. AB - Several important functions have been assigned to the receptor for urokinase-type plasminogen activator, uPAR. As implied by the name, uPAR was first identified as a high affinity cellular receptor for urokinase plasminogen activator (uPA). It mediates the binding of the zymogen, pro-uPA, to the plasma membrane where trace amounts of plasmin will initiate a series of events referred to as "reciprocal zymogen activation" where plasmin converts pro-uPA to the active enzyme, uPA, which in turn converts plasma membrane-associated plasminogen to plasmin. This is an efficient machinery to generate broad-spectrum proteolytic activity which is spatially restricted to the plasma membrane, since plasmin that diffuses away from the plasma membrane is rapidly inactivated by circulating inhibitors (i.e., alpha 2-antiplasmin). The system is controlled by a series of plasminogen activator inhibitors (PAIs), most importantly PAI-1 and PAI-2, providing means of temporally restricting the process of plasminogen activation. In addition to its role in plasminogen activation, compelling evidence has demonstrated a role for uPAR in cell-cell and cell-extracellular matrix adhesion, both directly and indirectly. uPAR is directly involved in binding to the extracellular matrix molecule, vitronectin, and the affinity of this binding is increased when uPAR is occupied by (pro-)uPA. A more indirect but presumably very important role of uPAR in cell adhesion seems to be mediated through interactions between uPAR and beta 1- or beta 2-integrins. It has been demonstrated that uPAR may bind physically to integrins in a reversible manner. The interaction seems to be of functional importance since the affinity of the integrin for its corresponding ligand is modulated by the association of integrin with uPAR. In some experimental setups uPAR has been shown to reduce the affinity of the associated integrin for certain ligands, while other experimental systems have demonstrated an increased affinity of the interaction between integrin and ligand after binding of uPAR to the integrin. Finally, uPAR has also been shown to participate in signal transduction events. Since uPAR is not a transmembrane molecule but belongs to the group of proteins that are tethered to the plasma membrane via a glycosyl phosphatidylinositol anchor, association with a transmembrane adaptor is required for transmission of signals via uPAR. Integrins may serve as such signal transducers, and indeed uPAR has been shown to be associated in the plasma membrane with complexes of integrins and (phosphorylated) tyrosin kinases suggesting a role for these complexes in transmembrane transmission of signals via uPAR. In the hematopoietic system it has been shown that urokinase-type plasminogen activator (uPAR) is expressed as a differentiation antigen on cells of the myelomonocytic lineage and as an activation antigen on monocytes and T lymphocytes. Neutrophils contain intracellular reservoirs of uPAR that are translocated to the plasma membrane upon activation, and neutrophils from patients with the rare blood disease paroxysmal nocturnal hemoglobinuria (PNH) that fail to express glycosyl-phosphatidylinositol-anchored proteins including uPAR, show a very significantly reduced transmigration over an endothelial barrier. Cell-associated plasminogen activation by PNH-affected neutrophils is severely impaired, and it has been proposed that this may be causally related to the propensity for thrombosis in PNH. The pattern of expression of uPAR in hematological malignancies mirrors the expression by normal blood and bone marrow counterparts with some exceptions (differentiated myeloid leukemias are positive, undifferentiated myeloid may be negative and the majority of lymphoid leukemias and lymphomas are negative). The potential clinical relevance of uPAR expression in leukemias and lymphomas has not been determined. PMID- 9402653 TI - Dendritic cell development: multiple pathways to nature's adjuvants. AB - Dendritic cells are a system of bone marrow-derived antigen-presenting cells specialized for interaction with T lymphocytes and essential for initiating primary T cell immune responses. Recent investigation indicates that dendritic cells are of diverse origin, with at least two types of myeloid precursors and a lymphoid precursor implicated in their generation. Mature dendritic cell subtypes, while sharing the capacity to activate T cells, show additional functional specialization. Some dendritic cells are equipped with additional mechanisms to regulate the response of the T cells they activate, while others are able to interact with B cells and modify B cell responses. PMID- 9402654 TI - Induction of c-kit molecules on human CD34+/c-kit < low cells: evidence for CD34+/c-kit < low cells as primitive hematopoietic stem cells. AB - c-kit, a receptor for stem cell factor, has been widely accepted as a distinctive marker for hematopoietic stem cells. However, the level of c-kit expression on pluripotent hematopoietic stem cells is still controversial in mice and humans. We purified CD34+/c-kit < low cells (phenotypically c-kit-negative but only detectable at the message level) from human cord blood and examined their maturational steps in relation to the expression of c-kit molecules. When the CD34+/c-kit < low cells were cultured with cytokines (flt 3 ligand, interleukin 6 and interleukin 7) plus immobilized anti-CD34 monoclonal antibody (to crosslink CD34 molecules), c-kit molecules were clearly induced within 24 h. The c-kit expression gradually increased until day 8. When CD34+/c-kit(low) or CD34+/c-kit+ cells that had been induced from CD34+/c-kit < low cells were resorted and recultured using a methylcellulose culture system, they showed the same colony forming ability as the freshly isolated CD34+/c-kit(low) or CD34+/c-kit+ cells, respectively. Furthermore, CD34+/c-kit < low cells have a similar hematopoietic potential to CD34+/c-kit(low) cells in assays for long-term culture initiating cell and colony-forming unit culture generated from long-term cultures. These findings suggest that CD34+/c-kit < low cells mature into CD34+/c-kit(low) and CD34+/c-kit+ cells, and acquire the reactivity to various humoral hematopoietic stimuli. Moreover, CD34+/c-kit < low cells showed a low level of rhodamine 123 retention, suggesting that CD34+/c-kit < low cells have multidrug resistance. Therefore, the CD34+/c-kit < low cells without colony-forming unit-granulocyte erythroid-macrophage-megakaryocyte activity are also a pluripotent hematopoietic stem cell population, and the expression of c-kit on c-kit < low cells is the first maturational step of hematopoiesis. PMID- 9402655 TI - Intrathymically injected hemopoietic stem cells can differentiate into all lineage cells in the thymus: differences between c-kit+ cells and c-kit < low cells. AB - To investigate whether hemopoietic stem cells (HSCs) can differentiate into all lineage cells even in the thymus, we injected two types of HSCs (c-kit+ and c-kit < low cells) obtained from C57BL/6 Ly5.1 mice directly into the thymus of 7.5 Gy irradiated C57BL/6 Ly5.2 mice. When c-kit < low cells (low density/lineage-/CD71 /major histocompatibility complex class I high/Sca-1+/Thy-1low/ c-kit < low) were injected, donor-derived (Ly5.1) cells were detected on day 8 after intrathymic (i.t.) injection, and the number reached a maximum on day 24 after injection. Granulocytes and macrophages were also detected on day 8 after injection. However, B220+ B cells were observed on day 13. Eighteen days after i.t. injection, the injected lobes showed red color due to the synchronous development of erythroid cells. Histological studies revealed the development not only of erythroid lineage cells but also of megakaryocytes in the thymus. In contrast, when c-kit+ cells were injected, a significant number of donor-derived cells were detected on day 5 after i.t. injection (three days earlier than in the case of c kit < low cell injection). The differentiation into erythroid lineage cells was also observed six days earlier than when c-kit < low HSCs were injected. These findings suggest that c-kit < low HSCs are more primitive than c-kit+ HSCs, although both can differentiate into all lineage cells after i.t. injection. PMID- 9402656 TI - IL-6 interferes with stimulation of HPP-CFC and large CFU-Mk in conjunction with cytokine combinations from primitive murine marrow cells. AB - The growth-promoting activities of interleukin 6 (IL-6) in combination with early acting hematopoietic factors, i.e., stem cell factor (SCF) and interleukin-1 alpha (IL-1 alpha), on primitive hematopoietic and megakaryocyte progenitors (high proliferative potential colony-forming cells [HPP-CFC] and colony-forming units-megakaryocyte [CFU-Mk], respectively) from 5-fluorouracil (5-FU)-treated murine bone marrow cells (BMC) were evaluated in serum-free fibrin clot cultures. IL-6 in combination with SCF and IL-1 induced an irregular and abortive hematopoiesis characterized by a reduction in colony size of at least 50% over those stimulated by SCF + IL-1 + IL-3 and an inability to continue growth to day 12. Moreover, IL-6 in combination with the early-acting factors, SCF and IL-1, had no effect on the formation of HPP-CFC. IL-6 is synergistic with SCF + IL-1 on day 7 CFU-Mk but did not stimulate large day 12 CFU-Mk. Our results suggest that, in the absence of serum, IL-6 prevents the continued proliferation of early hematopoietic and megakaryocytic progenitors initiated by SCF + IL-1 + IL-3. Optimization of cytokine combinations for use in ex vivo expansion of marrow progenitors, either for stem cell transplants or gene therapy, must consider not only the number of colonies but their size, as well as the contributions of serum components. PMID- 9402657 TI - Long-term cytokine production from engineered primary human stromal cells influences human hematopoiesis in an in vivo xenograft model. AB - Human hematopoiesis can be supported in beige/nude/ XID (bnx) mice by coinjection of human bone marrow stromal cells engineered to secrete human interleukin 3 (HuIL-3). The major limitation is a total absence of human B cell development in the mice, which could be due to supraphysiological levels of HuIL-3 in the circulation. In an effort to obtain human B lymphoid, as well as T lymphoid and myeloid cell development in the mice, CD34+ cells were coinjected with human marrow stromal cells engineered to secrete human IL-2, IL-7, stem cell factor or FLT3 ligand, +/- IL-3. No single factor other than IL-3 supported sustained human hematopoiesis in the mice, although cytokines were expressed for four to six months post-transplantation. Production of both HuIL-3 and IL-7 in the mice supported extrathymic development of human T lymphocytes, but no B cells, myeloid cells, or clonogenic progenitors were detected. Human B cells were not produced from CD34+ cells in the bnx mice under any condition tested. Another limitation to the bnx/Hu system is a lack of maturation of human red blood cells, although BFU-E are maintained. Stromal cells secreting human erythropoietin and IL-3 were cotransplanted into mice with HuCD34+ cells and an increase in hematocrit from 40%-45% to 80%-85% resulted, with production of human and murine red blood cells. Unfortunately, all mice (n = 9) suffered strokes, displayed paralysis and died within three weeks. The bnx/Hu cotransplantation model provides an interesting system in which to study human hematopoietic cell differentiation under the influence of various cytokines. PMID- 9402658 TI - Inhibitory action of the peptide AcSDKP on the proliferative state of hematopoietic stem cells in the presence of captopril but not lisinopril. AB - The effect of Angiotensin I-converting enzyme (ACE) inhibitors on their own and in combination with the peptide AcSDKP on the proliferation of hematopoietic stem cells has been investigated. Hematopoietic stem cells from murine bone marrow induced into cell cycle following exposure to 2 Gy gamma-irradiation were incubated in vitro for up to 24 h in the presence of medium, captopril/lisinopril, AcSDKP, and AcSDKP with either ACE inhibitor. Hematopoietic stem cells were monitored using the high proliferative potential-colony forming cell-1 (HPP-CFC-1) population cloned in the presence of human IL-1 beta, murine IL-3, and murine M-CSF. No significant inhibitory effect was observed in the presence of AcSDKP on its own and AcSDKP in combination with lisinopril. However, there was a significant inhibition of stem cell cycling when AcSDKP and captopril were combined. This suggests that captopril inhibits AcSDKP breakdown better than lisinopril. The combination of AcSDKP and captopril also had an inhibitory effect on cell recruitment into S phase. The fact that a combination of AcSDKP and captopril switches cycling hematopoietic stem cells out of cycle indicates the importance of the N-active catalytic site of ACE in AcSDKP hydrolysis in vitro. Thus, AcSDKP in combination with appropriate ACE inhibitors may be of use in regulating the proliferation of hematopoietic stem cells in vitro. PMID- 9402659 TI - Requirement of major histocompatibility complex-compatible microenvironment for spleen colony formation (CFU-S on day 12 but not on day 8). AB - To clarify major histocompatibility complex (MHC) restriction between hematopoietic stem cells (HSCs) and microenvironments, T cell-depleted bone marrow cells (BMCs) were transplanted into MHC-compatible and MHC-incompatible recipients. A significantly larger number of spleen colony-forming units (CFU-S) on day 12 were noted in MHC-compatible recipients, while only a small number were observed in MHC-incompatible recipients. There was, however, no significant difference in CFU-S counts on day 8 between the two groups. A large number of CFU S counts on day 12 were also observed in F1 hybrid recipients, as seen in syngeneic recipients. The decrease in CFU-S counts on day 12 in MHC-incompatible recipients was also observed even after in vivo abrogation of T and NK cells. The difference in CFU-S counts on day 12 became more prominent when HSC-enriched cells were transferred. These results suggest that an MHC restriction exists between pluripotent HSCs (P-HSCs) and spleen microenvironments. Furthermore, experiments using B10. A recombinant strains revealed that H-2D and S loci play a crucial role in the MHC restriction. The experiments of serial transplantation suggest that the differentiation and proliferation of P-HSCs are inhibited in MHC incompatible microenvironments. It is therefore likely that the MHC-compatible microenvironment is essential to the differentiation and proliferation of P-HSCs. PMID- 9402660 TI - Response of the cellular immune system to cardiopulmonary bypass in vivo. AB - Cardiopulmonary bypass (CPB) is known to induce an inflammatory response. Previous studies reported an impairment of the cellular immune response with activation of neutrophils and changes in lymphocyte subpopulations. The objective of the present study was to investigate the effect of CPB on leukocyte activation in vivo. In 27 patients undergoing coronary artery bypass grafting, the quantitative and the qualitative response of leukocyte populations to CPB was analysed pre-, intra-, and postoperatively using flow cytometry. A significant increase in leukocyte counts was detected during CPB, resulting in a marked leukocytosis postoperatively. The total number of lymphocytes peaked in the early phase of CPB, followed by a significant decrease, mainly due to a loss in B and cytotoxic T lymphocytes. In contrast, the lymphocytopenia observed 8 h after protamin administration was mainly caused by a drop in the population of helper T lymphocytes. Activation of distinct cell populations could be detected during and following CPB. The results indicate an influence of CPB on the cellular immune system, however an immuno-suppression was detectable only transiently. PMID- 9402662 TI - Tachyarrhythmias following coronary artery bypass graft surgery: epidemiology, mechanisms, and current therapeutic strategies. AB - The distribution pattern of perioperative complications of coronary artery bypass graft surgery (CABG) is currently changing, due to the fact that it has become a routine operation with an increasing proportion of patients older than 70 years. Supraventricular and ventricular tachyarrhythmias are among the most commonly observed postoperative adverse events following CABG. The aim of this review is to give an update on epidemiology and mechanisms of postoperative arrhythmias and to present current diagnostic tools and therapeutic strategies. PMID- 9402661 TI - Improved myocardial protection using continuous coronary perfusion with normothermic blood and beta-blockade with esmolol. AB - Continuous coronary perfusion with warm beta-blocker-enriched blood has been suggested as an alternative to cardioplegic arrest for myocardial protection during coronary artery surgery. The purpose of the present work was 1.) to experimentally investigate this technique using an animal model, and 2.) to clinically apply this alternative myocardial protection technique and compare it to standard crystalloid cardioplegia in a controlled study. We placed 6 dogs on CPB and 6 dogs on a biventricular assist device and created "beta-blocker-induced cardiac surgical conditions" by suppressing myocardial chronotropy and inotropy with systemic infusion of the ultra-short acting beta-blocker esmolol. For the clinical study we randomized 60 coronary artery surgery patients to receive either crystalloid cardioplegia (Bretschneider HTK) or selective continuous coronary perfusion via the aortic root with warm esmolol-enriched CPB blood. In the experimental study we found that continuous coronary perfusion with warm esmolol-enriched blood avoided myocardial ischemia and minimized myocardial edema, thus completely preserving cardiac performance. Our clinical data showed the alternative technique to be superior to standard crystalloid cardioplegia in terms of both functional and structural myocardial protection. The concept of beta-blocker-induced cardiac surgical conditions is a useful alternative for myocardial protection during coronary artery surgery and may be particularly beneficial for severely compromised hearts. PMID- 9402663 TI - Open heart interventions in premature low- and very-low-birth-weight neonates: risk profile and ethical considerations. AB - In premature, very-low-birth-weight (VLBW) neonates, complex cardiac malformations can be successfully repaired under conditions of cardiopulmonary bypass. However, due to the immaturity of organ systems, these patients are exposed to a specific risk resulting from noxious effects of extracorporeal circulation, especially on the central nervous system. Two premature neonates with low and very low birth weight of 1160 g and 1650 g, were operated on using cardiopulmonary bypass for severe pulmonary artery stenosis and truncus arteriosus communis type II, respectively. The neonate with pulmonary valve stenosis survived, but at 2-year-follow-up examination motoricity retardation as a result of cerebral immaturity-related changes was evident. The other neonate died suddenly on the fifth postoperative day of a massive intracranial haemorrhage. Due to the fact that the natural history of VLBW children is a priori characterized by a high incidence of major neurological handicaps, open heart surgery may by improving survival chances contribute to an increased incidence of mentally handicapped children. PMID- 9402664 TI - Long-term follow-up of patients with operative stabilisation of a flail chest. AB - The outcome is reported of patients after external chest wall stabilisation for respiratory insufficiency due to a traumatic flail chest. Since 1990, all patients with a flail chest causing respiratory insufficiency despite peridural analgesia and without further reason for prolonged mechanical ventilation underwent osteosynthesis of the chest wall using the AO-technique with 3.5 mm thick reconstruction plates, and were prospectively followed-up by use of clinical and radiological evaluation. 23 patients underwent external chest wall fixation between 1990 and 1996 and were followed for a mean time of 28 months. 2 patients died after the operation, giving a 30-day-survival rate of 91.3% 21 patients survived and were extubated and transferred to the ward after a mean time interval of 3.9 and 7.8 days, respectively. 95% of the survivors revealed a 100% working capacity at assessment and 86% returned to preoperative sports activities without complaining of chest wall or shoulder girdle pain or dysfunction. External chest wall fixation appears to be an attractive alternative to prolonged intubation and mechanical ventilation for selected patients with flail-chest respiratory insufficiency despite peridural analgesia, providing they do not require prolonged intubation for other reasons. PMID- 9402665 TI - Myxoma complex associated with transient hyperthyroidism: are diseases of the thyroid part of the complex? AB - The myxoma complex is a very rare disease. We report on a 31-year-old woman who suffered a hyperthyreosis, which resolved spontaneously. A few months later she developed a Cushing's syndrome, caused by a primary adrenocortical nodular dysplasia. A bilateral adrenalectomy had to be performed. 15 months after this operation she developed a cerebral infarction caused by an embolus from a left atrial myxoma. Our patient is a further case of Carney's syndrome. The aspect of the hyperthyreosis is interesting: Perhaps disease of the thyroid represents a new element of Carney's syndrome. PMID- 9402666 TI - Sweet's syndrome and associated sarcoidosis--a rare clinical case. AB - Sweet's syndrome is frequently idiopathic; however, it has been associated with malignancy in up to 20% of reported cases. We report a case of concurrent presentation of Sweet's syndrome with sarcoidosis. Although this association has been infrequently described we feel that this could be an alternative diagnosis in patients with Sweet's syndrome who present with mediastinal lymphadenopathy with or without erythema nodosum. The relevant literature on this subject is reviewed. PMID- 9402667 TI - Cardiac hydatid cyst causing massive pulmonary embolism. AB - Cardiac hydatid cyst is a rare parasitic disease. Since it may be associated with fatal complications, early diagnosis and treatment of a cardiac hydatid cyst is very important. We present a case with hydatid cyst localized in the right atrium and bilaterally in the lungs, and embolized pulmonary arteries bilaterally. The right atrial cyst localized on the interatrial septum was removed using cardiopulmonary bypass and the cyst in the right pulmonary artery was extracted by an embolectomy catheter. The patient died of pulmonary hypertension and pulmonary insufficiency three months postoperatively. PMID- 9402668 TI - Constrictive pericarditis caused by an old hematoma. AB - A 46-year-old woman was admitted for surgery after diagnosis of a constrictive pericarditis. She had suffered from acute pericarditis and undergone pericardiocentesis to relieve pericardial effusion several times 10 years previously. Cardiac catheterization showed elevated mean right-atrial and pulmonary wedge pressures. Chest CT revealed only a calcified mass in the retrosternal space. At surgery, there was no severe adhesion between the pericardium and the epicardium, and the pericardium was thin and almost normal except over the right artio-ventricular groove, where a hard mass with an atheromatous content adhered to the epicardium. After resection of the mass, central venous pressure and pulmonary wedge pressure decreased. The mass was found to be an old hematoma by pathological examination. The patient was subsequently discharged from hospital in good condition. PMID- 9402669 TI - Primary tracheal fibrosarcoma in a child: a case of tracheal resection under ECMO support. AB - We report a case of severe airway obstruction due to endotracheal fibrosarcoma in a 3-year-old boy. Successful tracheal resection and reconstruction was performed under extracorporeal membrane oxygenation support. A solid, elastic hard tumor with a smooth surface was attached by a tiny stalk structure to the membranous part of the lower trachea. Histological findings of the tumor were consistent with infantile fibrosarcoma, showing proliferation of spindle cells forming interlacing patterns. PMID- 9402670 TI - Epithelioid hemangioendothelioma presenting as a chest wall tumor. AB - We report a case of epithelioid hemangioendothelioma presenting as a chest wall tumor. The patient had invasive bladder carcinoma and a soft-density mass protruding into the left thoracic cavity from the lateral chest wall on a computed tomographic scan. Percutaneous needle biopsy was performed to obtain a definite diagnosis of the chest wall tumor. Because of an intrathoracic hemorrhage after the procedure, the patient underwent an emergency thoracotomy and excision of the mass with adjacent structures. Pathologic examination demonstrated the mass to be a subpleural epithelioid hemangioendothelioma. This rare tumor has never been reported previously as arising from the chest wall. PMID- 9402671 TI - "Reversed" latissimus dorsi musculocutaneous flap for closure of large bronchopleural fistula. AB - The patient was a 54-year-old male with diabetes mellitus and liver abscess perforating into the right lung through the diaphragm. After right lower lobectomy of the lung, S3-segmentectomy of the liver, and debridement of the subdiaphragmatic abscess a bronchopleural fistula appeared. After open-drainage thoractomy, secondary operation for closure of a large bronchopleural fistula and obliteration of the empyema cavities was performed with a "reversed" latissimus dorsi musculocutaneous flap. PMID- 9402672 TI - Pulmonary metastasis from a basal-cell carcinoma of the retroauricular region. AB - Basal-cell carcinoma of the skin is a common facial neoplasm, usually regarded as benign. It is also called basalioma. Distant metastasis is very rare and may involve the brain, lung, and bones. We report a 74-year-old white male who was admitted to our hospital with cough and fever. Chest radiograph revealed an opacity of 2 x 1 cm in diameter in the upper lobe of the right lung. Bronchoscopy and thoracic fine-needle aspiration could not establish a diagnosis. Therefore the patient underwent right thoracotomy and wedge excision of the lesion. Histologic evaluation was consistent with pulmonary metastasis of a facial basal cell carcinoma. The patient recovered uneventfully from surgery and is well 5 years after the operation. According to the English literature the median survival of patients with metastatic basal-cell carcinoma is 10 months. The clinical features, pathology, and treatment of this rare entity are discussed. PMID- 9402673 TI - Bronchogenic carcinoma following pulmonary aspergilloma. AB - We have observed two cases of bronchogenic carcinoma following pulmonary aspergilloma among 25 cases of pulmonary aspergilloma. Patients with a pulmonary aspergilloma may be susceptible to bronchogenic carcinoma. PMID- 9402674 TI - Doubly angled pleural drain circumventing the transcostal route relieves pain after cardiac surgery. AB - Standard pleural drainage after cardiac surgery is accomplished through the intercostal space and the divided parietal pleural, often causing severe additional chest pain. To circumvent this route of insertion a doubly angled polyvinyl chloride drain was developed which can be placed via the median approach through the rectus abdominis muscle just beside the anterior mediastinal drains without irritation of the heart and parietal pleura into the phrenico costal sinus. PMID- 9402675 TI - Surgical treatment of aortic root abscess with damage of the mitral valve apparatus. AB - A 35-year-old man with an aortic root abscess successfully underwent aortic valve replacement and mitral valve repair using a prosthetic valve with a skirt which was secured to the sewing cuff. This is a useful technique for reinforcement of a fragile annulus and prevention of annular detachment. A further useful technique is to infuse normal saline from a Foley catheter inserted into the aortic root to estimate mitral valve coaptation. PMID- 9402676 TI - Three-dimensional contrast-enhanced MR angiography. AB - Three-dimensional contrast-enhanced magnetic resonance (MR) angiography is a relatively new technique that uses the transient shortening in blood T1 following the intravenous injection of gadolinium chelates to image blood vessels irrespective of flow. For many applications, 3D contrast-enhanced MR angiography is developing into a safe, fast, and cost-effective alternative to conventional diagnostic angiography. One of its biggest advantages over other MR angiography techniques (and CT angiography) is the ability to image in a plane parallel to the vessel of interest. This feature, combined with the inherent properties of a 3D gradient refocused sequence, make 3D contrast-enhanced MR angiography intrinsically fast, high resolution and free from saturation and turbulence related artifacts. This article is designed to familiarize the reader with the theory of 3D contrast-enhanced MR angiography and the application of the technique to different vascular territories. Contrast agents, relaxation effects, contrast bolus effects, pulse sequences, artifacts, and post-processing, as well as the present state of thinking with regard to optimal contrast injection timing/detection and Fourier space mapping are discussed. Patient preparation and techniques and imaging parameters for body applications of gadolinium-enhanced MR angiography, including aorta, renal arteries, mesenteric arteries, portal venous system, pelvis and legs, pulmonary arteries, and carotid arteries are included. PMID- 9402677 TI - MR angiography of the head and neck. AB - A review of the basic physics and techniques for acquiring and evaluating magnetic resonance angiograms is provided, including time-of-flight and phase contrast techniques. Magnetic resonance (MR) angiography is becoming a routine method of evaluating carotid bifurcation atherosclerotic disease in both a screening and diagnostic capacity. The expanding clinical utility of MR angiography in the detection of intracranial aneurysms, characterization of arteriovenous malformations, and evaluation of intracranial atherosclerotic disease are also reviewed. Furthermore, MR angiography allows for the noninvasive diagnosis of arterial dissection. Magnetic resonance venography also allows the confirmation of the previously elusive and likely underdiagnosed entity of cerebral venous thrombosis. PMID- 9402678 TI - MR angiography with three-dimensional MR digital subtraction angiography. AB - We have developed a time-resolved, contrast-enhanced, volume-imaging technique for magnetic resonance (MR) angiography, known as three-dimensional (3D) MR digital subtraction angiography (DSA). This technique greatly improves MR angiogram quality because it combines the injection of a contrast agent with the ability to image the temporal passage of this agent and, thereby, obviates the need for timing scans or other complicated synchronization schemes. Three dimensional MR DSA also represents a potential improvement in the sense that, relative to DSA and computed tomography (CT) angiography, the contrast agent is less toxic. Additionally, unlike CT angiography, images may be acquired during the passage of the contrast agent. Therefore, 3D MR DSA shows the sequential passage of contrast through the arterial and venous system, followed by uptake in various organs. Unlike conventional DSA, 3D MR DSA imaging acquires full volume datasets, which allows subsequent reprojection and reformatting. Because images are obtained at approximately 2-6 s time intervals using a temporal aperture on the order of several seconds, motion (such as respiration) causes only a temporary disruption of image quality, similar to that observed in MR fluoroscopy. These temporal characteristics also make the proposed sequence insensitive to variations in the shape and timing of the contrast-pass curve. Although the individual time-resolved images will have somewhat decreased signal to-noise ratio (SNR) relative to nontime-resolved scans collected in the same acquisition time, the SNR improvement due to the gadolinium appears to accommodate this trade-off. Additionally, if motion between successive images is small, then the full suite of temporal processing schemes, previously investigated in connection with DSA and time-resolved two-dimensional (2D) MR, such as mask mode subtraction, simple matched filtering and Eigen filtering, can be used to obtain composite images. These derived images generally have an increased SNR or negligible venous signal if an arterial-phase image is not obtained in the early time-resolved images. In summary, 3D MR DSA will significantly advance MR angiography because of the following intrinsic advantages: (1) improved signal-to-noise, (2) scan orientation may be chosen independently of the direction of blood flow, (3) uniform vascular signal, even from regions of complex flow, (4) minimization of motion artifacts, (5) greatly reduced sensitivity to variation in the shape and timing of the contrast bolus, (6) ability to be reformatted or reprojected, and (7) ability to apply a variety of temporal postprocessing techniques. PMID- 9402679 TI - Fluid-attenuated inversion recovery (FLAIR): clinical prospectus of current and future applications. AB - A relative weakness of the traditional spin-echo technique, and particularly of the newer "FAST" or "TURBO" spin-echo sequences, has been diminished conspicuousness of lesions affecting the peripheral cortical mantle or those located in the periventricular region. This is a consequence of partial volume effects and high cerebrospinal fluid (CSF) signal adjacent to pathologic regions. Fluid-attenuated inversion recovery (FLAIR) is a magnetic resonance imaging (MRI) sequence that produces strong T2 weighting, suppresses the CSF signal, and minimizes contrast between gray matter and white matter. This effect produces images with significantly increased lesion-to-background CSF contrast and enhances the visibility of lesions as well as their detectability, particularly in the peripheral subcortical and periventricular regions. Applications are evolving, though preliminary reports highlight the superiority of FLAIR in the evaluation of infarction, multiple sclerosis, metastatic disease, tuberous sclerosis, and, possibly, subarachnoid hemorrhage. Early reports also address the application of FLAIR to imaging of the spinal cord. Modified versions of FLAIR are currently being developed; these modifications will further shorten acquisition times and eliminate pulsation artifacts. FLAIR may ultimately supplant conventional spin-echo imaging in routine MR screening of the brain. PMID- 9402680 TI - Concerns regarding the current paradigm for chronic allograft rejection. PMID- 9402681 TI - Role of macrophages in vascular tissue remodelling. PMID- 9402682 TI - Cell-cell interactions in vessel assembly: a model for the fundamentals of vascular remodelling. PMID- 9402683 TI - Extracellular matrix modulation of vascular cell behaviour. PMID- 9402684 TI - Apoptosis in vascular disease. PMID- 9402685 TI - Cytokine mRNA expression in tolerant heart allografts after immunosuppression with cyclosporine, sirolimus or brequinar. AB - We sought to examine the impact of the preferential activation of Th2 cells on the induction and maintenance of a tolerant state in heart allograft rat recipients treated with a short course of cyclosporine (CsA), sirolimus (SRL) or brequinar (BQR). A quantitative polymerase chain reaction (PCR) method was used to measure the levels of cytokine mRNAs, namely interferon (IFN)-gamma and interleukin (IL)-2 in T helper 1 (Th1) cells and IL-4, IL-5 and IL-10 in Th2 cells. Our main findings were that on day 5 postgrafting allografts from untreated recipients had increased levels of IFN-gamma (216 +/- 119 fg), IL-2 (449 +/- 75 fg), IL-4 (6.2 +/- 1.3 fg), IL-5 (34.8 +/- 9.3 fg) and IL-10 (1554 +/ 184 fg) mRNAs compared with normal hearts. CsA reduced the levels of IFN-gamma, IL-2, IL-5 and IL-10, but not IL-4, mRNAs. SRL did not affect the expression of cytokine mRNAs. BQR decreased the levels of IFN-gamma, IL-2 and IL-10, but not IL 5 or IL-4 mRNAs. Compared with grafts from untreated recipients, those from CsA- or BQR-treated tolerant hosts (day 100) displayed undetectable IL-2 mRNA levels, and reduced levels of IFN-gamma, IL-4 and IL-10 mRNAs. In fact, the patterns of cytokine mRNA expression in grafts from CsA- and BQR-treated tolerant hosts were similar to those of normal hearts. Grafts from SRL-treated tolerant hosts merely showed slightly increased Th2 cell activity. In conclusion the selective activation of Th2 cells is not absolutely required for induction or maintenance of tolerance. PMID- 9402686 TI - Interleukin-12 p40 m-RNA expression in human kidney allograft biopsies. AB - Interleukin-12 (IL-12) is a heterodimeric cytokine implicated in the early differentiation of naive T-lymphocytes into the Th1 subset. IL-12 is important for induction of the cellular immune response against viruses, intracellular parasites and neoplasms. Its role in alloresponsiveness has not been fully elucidated. Preliminary data in the literature point toward the prevalence of Th1 lymphocytes in processes of allograft rejection. In attempt to further investigate the expression of this cytokine during episodes of cellular rejection of renal allografts, we searched for IL-12 message in human kidney allograft biopsies using the reverse transcriptase-polymerase chain reaction technique. Twenty-three allograft core biopsies from 19 patients were obtained percutaneously for clinical indications in 18 cases, and as part of an investigational protocol in five cases. A portion of the tissue was used for RNA extraction using the guanidium-thiocyanide phenol-chloroform method. Histology was performed on the remaining core material. Ten mg of total RNA were used for reverse transcription. PCR of the c-DNAs was done for 40 cycles using primers for the p40 subunit of IL-12 and GAPDH which was used as a control. PCR products were photographed after electrophoresis, transferred to a nylon membrane and hybridized with a radiolabelled cloned human IL-12 p40 1 kb c-DNA fragment. Autoradiographies were developed after 20-min exposure. All samples were run in triplicate. IL-12 p40 m-RNA was expressed in all 17 biopsies showing acute cellular rejection as well as in all three biopsies showing focal interstitial fibrosis. No message was found in the presence of normal allograft histology. This is the first in vivo report of IL-12 p40 subunit m-RNA expression during renal allograft rejection in humans. The role of this Th1 cytokine in the alloresponse deserves further investigation. PMID- 9402687 TI - Single dose anti-CD4 monoclonal antibody for induction of tolerance to cardiac allograft in high- and low-responder rat strain combinations. AB - Repeated administration of monoclonal antibodies (mAb) directed against the CD4 lymphocyte receptor may induce specific, long-lasting unresponsiveness to fully MHC-mismatched cardiac allografts in rats without additional immunosuppression. We assessed the effect of a single dose of murine anti-rat depleting anti-CD4 mAb (OX-38) on allograft survival in high- and low-responder rat strain combinations. Isogenic strains of DA (RT1av1), PVG (RT1c), AUG (RT1c), and WF (RT1u) rats were used. Recipients in antibody treated groups were given one dose of 5 mg/kg OX-38 mAb on the day of transplant, a dose which was shown to effectively deplete (or block) circulating CD4+ T cells. Other groups were treated for 10 days with cyclosporin A (CsA) and/or Linomide, a novel immunomodulator, which is the first compound able to fully eliminate the effect of CsA in the rat cardiac allograft model. The DA strain was identified as a low-responder to the allogeneic haplotype RT1c (PVG or AUG), but not to RT1u (WF), and developed true tolerance following RT1c grafting and OX-38 or low-dose CsA (5 mg/kg) induction, as verified by the response to retransplantation of a graft from the same donor strain or a third-party challenge. PVG recipients of DA grafts were characterized by high response and only modest (OX-38; median 9.5 days) or moderate (CsA; 23.5 days) prolongation of graft survival. Contrasting graft survival results were obtained in the low-responder combination, either very early rejection (at 10 days) or permanent graft survival (> 100 days). Linomide challenge affected CsA treatment in the high-responder combination but not tolerance induction in the low-responder combination, or the effect of OX-38. It was concluded that in rat heart transplantation a single-dose anti-CD4 mAb therapy may induce permanent donor-specific unresponsiveness in a low-responder strain combination, and that anti-CD4 mAb seems to be unique among immunosuppressive agents while being resistent to challenge by Linomide. PMID- 9402689 TI - Serum soluble human leucocyte antigen class I in paediatric liver transplantation with live, related donors. AB - Serum soluble human leucocyte antigen (HLA) class-I is a useful marker for predicting immunological events in organ transplantation. In cadaver liver transplant cases it is especially the case that high amounts of soluble HLA-I are excreted from the grafts. In Japan, almost all liver transplants have been performed from living parent donors to their children. Therefore, it is interesting to know how soluble HLA-I changes in relation to clinical course. As part of this study we first examined serum concentrations of soluble HLA-I in 33 paediatric patients using enzyme-linked immunosorbent assay. Soluble HLA-I is composed of three different sized molecules (45, 39 and 34-36 kDa); then the change of distribution of these three molecules was demonstrated by Western blot analysis. When donor and recipient have different soluble HLA-I band patterns, the origin of the antigen can be assumed by this method. We found that in a comparison between pre- and post-transplants, the six out of eight (75%) patients that suffered episodes of acute rejection showed a significant elevation of soluble HLA-I, and all patients with infectious episodes had an elevated soluble HLA-I. Meanwhile, 10 out of 22 (45%) patients without any clinical complications still showed increased soluble HLA-I. The Western blot analysis showed that the soluble HLA-I molecules were considerably derived from the grafted liver, from one week to 24 months after grafting. In acute rejection, the band signals of donor origin were significantly increased. These signals were attenuated after immunosuppressive therapy. The grafted liver appears to contribute to the increase of soluble HLA-I following liver transplantation, and this increase is greater with the effects of the host immune system. PMID- 9402690 TI - Unexpected augmentation of mycophenolic acid pharmacokinetics in renal transplant patients receiving tacrolimus and mycophenolate mofetil in combination therapy, and analogous in vitro findings. AB - Mycophenolate mofetil (MMF) a potent immunosuppressive agent, has recently been approved for clinical use (CellCept) in renal transplant patients in combination with cyclosporine (CsA). With the expanded use of tacrolimus (Prograf) as well in renal transplant patients, there is a lack of pharmacokinetic studies clarifying drug interactions between the three agents. A pharmacokinetic study was performed on 18 stable renal transplant patients receiving MMF and tacrolimus together, and four control groups, one receiving tacrolimus alone, two receiving CsA, in combination with MMF (1.0 or 1.5 g bid), and one receiving CsA microemulsion (Neoral). Area-under-the-curve values were calculated for each drug to assess if there was a reciprocal effect on the respective bioavailability of each. In vitro, the immunosuppressive effect of trough level plasma from each patient group was studied using mixed lymphocyte culture (MLC), as well as MLC reactions spiked with various combinations of each drug. There was a minimal effect of MMF on tacrolimus pharmacokinetics. However, patients receiving tacrolimus and MMF displayed significantly higher levels (Cmin and area under the curve) of mycophenolic acid (MPA) than those receiving CsA (Sandimmune or Neoral) and the same dose of MMF (50.2 +/- 16.5 vs 32.1 +/- 16.7 micrograms h/ml AUC, p < 0.02). Equivalent MPA levels could be attained in patients receiving CsA if the MMF dose was increased by 50% (1.5 g bid). There were also significantly lower levels of the glucuronide metabolite of MPA (MPAG) (755 +/- 280 vs 1230 +/- 250 micrograms h/ml AUC, p = 0.02), suggesting a specific inhibition (either direct or indirect) of the conversion of MPA to MPAG in tacrolimus patients, as opposed to those receiving CsA. For each drug combination, there was a positive correlation between the plasma immunosuppressive effect seen in MLC assays and the MMF dose. In addition, trough plasma from patients receiving tacrolimus and MMF was significantly more MLC inhibitory than from those receiving CsA or CsA microemulsion and equivalent-dose MMF. Culture media containing MPA and tacrolimus equal to clinical therapeutic trough concentrations (10 ng/ml) were significantly more MLC inhibitory than CsA at equivalent clinical therapeutic trough concentrations (200 ng/ml) with equivalent MPA levels. These studies in renal transplant patients suggest that tacrolimus in combination with MMF may result in a greater degree of immunosuppression than may be anticipated. PMID- 9402691 TI - Antigen presentation by endothelium: heparin reduces the immunogenicity of interferon-gamma-treated endothelial cells. PMID- 9402688 TI - The role of natural anti-Gal alpha 1-3Gal antibodies in hyperacute rejection of pig-to-baboon cardiac xenotransplants. AB - Xenoreactive natural antibodies in humans and higher primates are directed predominantly at Gal alpha 1-3Gal. These antibodies are thought to initiate hyperacute rejection of porcine organ xenografts. The contribution of anti-Gal alpha 1-3Gal antibodies to the xenoractive natural antibody repertoire and to the initiation of hyperacute rejection was tested in a pig-to-baboon cardiac xenograft model. Anti-Gal alpha 1-3Gal antibodies were depleted from baboons by extracorporeal absorption of anti-Gal alpha 1-3Gal antibodies from plasma using columns with a matrix bearing Gal alpha 1-3Galb1-4GlcNAc. Specific removal of anti-Gal alpha 1-3Gal antibodies was achieved prior to transplantation as demonstrated by immunoassay. Porcine hearts were then transplanted into these baboons and the outcome of the transplants was analysed. Immunofluorescence revealed little deposition of baboon antibodies in the grafts. The porcine hearts did not undergo hyperacute rejection even though complement activity was approximately 90% of baseline at the time of transplantation. These findings demonstrate that anti-Gal alpha 1-3Gal antibodies constitute a major fraction of xenoreactive natural antibodies in primate blood and that these antibodies contribute significantly to the pathogenesis of hyperacute xenograft rejection. PMID- 9402692 TI - Production of IL-2 by hepatocytes in allografted liver of rats. PMID- 9402693 TI - Understanding the epidemiology of BSE. PMID- 9402694 TI - Multiple myeloma and Kaposi's sarcoma-associated herpesvirus--a paracrine model of tumorigenesis? PMID- 9402695 TI - To be a mutator, or how pathogenic and commensal bacteria can evolve rapidly. PMID- 9402696 TI - Mycobacterium tuberculosis: beyond genome sequencing. PMID- 9402697 TI - Pathogenicity islands: important but not unique factors contributing to Salmonella virulence. PMID- 9402698 TI - Toxins or signals in Dutch elm disease pathogenesis. PMID- 9402699 TI - Developmental biology of biofilms: implications for treatment and control. AB - Although of heterogeneous spatiotemporal and species compositions, all biofilms undergo certain common developmental events: organic molecules on the substratum can play a role in initial attachment, attached cells grow and additional cells attach from the bulk liquid. Biofilm growth is a four-dimensional (X, Y, Z and T) process similar to organ development. PMID- 9402700 TI - How rolling circle plasmids control their copy number. AB - Rolling circle DNA replication is inherently continuous and unregulated. This 'go for-broke' strategy works well for lytic phages but is suicidal for plasmids that must coexist with their host. Plasmids have consequently evolved elaborate copy number control systems that operate at the transcriptional, translational and post-translational levels. PMID- 9402701 TI - Why are hepadnaviruses DNA and not RNA viruses? AB - Virion assembly in hepadnaviruses is a two-step process leading to (1) the packaging of viral pregenomic RNA and reverse transcriptase into nucleocapsids and (2) the assembly of nucleocapsids with envelope components, which results in the formation of mature virus particles. Characteristically, both steps are intimately coupled to viral DNA synthesis. While assembly of nucleocapsids is coupled to the protein priming of reverse transcription, virion formation is linked to genome maturation. PMID- 9402702 TI - Filamentous growth in budding yeast. AB - The recent discovery that some laboratory strains of Saccharomyces cerevisiae are capable of limited filamentous growth has stimulated genetic analysis of dimorphism in this microorganism. The puzzle of why most strains are nonfilamentous is now resolved. Remarkably, a single point mutation with broad consequences separates these domesticated yeast from their wild ancestors. PMID- 9402703 TI - Are bacterial exotoxins cytokine network regulators? AB - Bacterial exotoxins are generally thought to act by damaging cells or altering cell metabolism. However, recent work has established that many exotoxins modulate eukaryotic cell cytokine synthesis. Cytokine induction may play a significant role in exotoxin action, and therapeutic targeting of cytokines could be beneficial in infectious diseases involving bacterial exotoxins. PMID- 9402704 TI - White collar proteins: PASsing the light signal in Neurospora crassa. AB - The filamentous fungus Neurospora crassa is an excellent paradigm for the study of blue light signal transduction. The isolation and characterization of the genes for two central regulators of the blue light response, white collar-1 and white collar-2, have begun to shed light on the mechanism of blue light signal transduction in fungi. These proteins are not only proposed to encode blue-light activated transcription factors but also to be elements of the blue light signal transduction pathway. PMID- 9402705 TI - Correlation between thoracic radiographic changes and remission/survival duration in 270 dogs with lymphosarcoma. AB - A retrospective study was undertaken wherein the medical records and thoracic radiographs of 270 dogs with lymphosarcoma were reviewed to determine the type and frequency of thoracic radiographic changes. Statistical evaluation of the relationship between radiographic, clinical and immunologic factors and the primary remission duration and survival times was performed using univariate and multivariate analysis. One hundred ninety-two dogs (71%) had some type of thoracic radiographic abnormality, including 80 dogs (29.6%) with pulmonary infiltrates and 164 dogs (64.4%) with thoracic lymphadenomegaly. Only T-cell phenotype (p = 0.0056 for survival, p = 0.0045 for remission) and the presence of cranial mediastinal lymphadenomegaly (p = 0.0005 for survival, p = 0.0129 for remission) were identified as having a significant negative correlation to both primary remission and survival duration by multivariate analysis. PMID- 9402706 TI - Radiographic imaging of otitis media and interna in pigs. AB - Middle and inner ear infections have been reported as a clinical entity in swine, other animal species and humans. In pigs, the anatomical-pathological and microbiological findings have been described. In this report, we describe radiographic findings in affected pigs. A total of 25 pigs with a head tilt and circling, as clinical signs of otitis media and interna, were examined. The majority were weaner-pigs with dyspnea or rhinitis. In radiographs, there was an increased opacity of the bulla tympanica, often accompanied by marginal destruction or thickening of the bulla wall. The radiographic findings confirmed the clinical diagnosis in each affected pig, but there were 5 false positive interpretations. PMID- 9402707 TI - Computed tomographic evaluation of comminuted middle phalangeal fractures in the horse. AB - Comminuted fractures of the middle phalanx have been well described in the horse. Choice of treatment, surgical planning and prognosis have traditionally been based upon evaluation of radiographs. However, the complex nature of comminuted fractures makes radiographic interpretation difficult. Computed tomography (CT) allows the production of cross-sectional images with spatial separation of structures which are superimposed on survey radiographs. This allows accurate assessment of the number and direction of fracture lines within the bone. In this paper we report the use of CT in the evaluation of 6 comminuted middle phalangeal fractures. Computed tomography is potentially useful in deciding the type of treatment, surgical planning and determining the prognosis. PMID- 9402708 TI - Magnetic resonance imaging of the lateral ventricles in beagle-type dogs. AB - Magnetic resonance (MR) imaging was performed on 21 presumed normal Beagle-type dogs. The size and symmetry of their lateral ventricles were evaluated and dogs were categorized on the basis of the percentage of their ventricular height (Vh) to brain height (Bh) and the ratio of the largest to the smallest ventricular area (rVA). Eleven dogs had lateral ventricles classified as normal sized (0-14% Vh/Bh) while 10 of 21 dogs had moderate enlargement (15-25% Vh/Bh) of one or both lateral ventricles. None of the dogs had severe lateral ventricular enlargement. Degree of asymmetry was also arbitrarily categorized on basis of rVA as normal to minimal (rVA < 1.5), mild (1.5 < rVA < 2.0), or severe (2.0 < rVA). Of the dogs having normal-sized lateral ventricles, six of eleven had symmetric, three of eleven had mildly asymmetric and two of eleven had severely asymmetric lateral ventricles. Of the dogs having at least one moderately enlarged lateral ventricles, five of ten had symmetric lateral ventricles, and two of ten had mild asymmetry and three of ten had severe asymmetry. Gender and body weight had no statistical relationship to lateral ventricle symmetry. Clinically insignificant ventricular enlargement and asymmetry was common in this group of Beagle dogs. PMID- 9402709 TI - Radiographic diagnosis: arachnoid cyst in a dog. PMID- 9402710 TI - Quantification of cerebral ventricular volume in English bulldogs. AB - Quantitative measurement of cerebral ventricle volume of eight English bulldogs was performed using magnetic resonance (MR) imaging. The mean ventricular volume was 14.8 ml. with a range of 8.6 ml.-38.1 ml. The mean ventricular volume of two beagles was 2.2 ml with a range of 0.7 ml.-3.7 ml. The percent of intracranial volume occupied by ventricle was found to be significantly larger in bulldogs (14.0%; S.D. = 7.9%) than in beagles (Range = 1.0-4.8%). The relationship between the percent of intracranial volume occupied by ventricle and measurements of body weight, age, sex, and various measures of skull anatomy of the bulldog was also determined. The relationship between ventricular volume and neurologic dysfunction was examined. There was a possible trend between high percent of intracranial volume occupied by ventricle and low body weight. This study will serve as a pilot study for examining the relationship between ventricular volume and neurologic disease in bulldogs. PMID- 9402711 TI - Appearance of canine abdominal tumors with magnetic resonance imaging using a low field permanent magnet. AB - The study was carried out to evaluate the applicability of magnetic resonance imaging (MRI) in detecting tumors in the abdomen of the dog. Abdominal ultrasound and MRI were performed on 8 dogs having a mass lesion on abdominal radiography. MR images were obtained in the transverse, sagittal and dorsal planes using T1- and T2-weighted spin echo pulse sequences. There was good visual correlation of the lesion site by MRI and ultrasonography (US). PMID- 9402712 TI - Retrospective ultrasonographic evaluation of adrenal lesions in 26 dogs. AB - A review was performed of ultrasonographic findings in 26 dogs with confirmed adrenal lesions. Adrenal shape, size, echogenicity, laterality, and the presence of vascular invasion were evaluated. Histopathologic diagnoses were obtained in all dogs. Adrenal lesions were confirmed as pheochromocytomas (9), adenocarcinomas (6), a poorly differentiated blastoma (1), bilateral adrenal metastases of a carcinoma (1), adenomas--one of which was bilateral--(4) and hyperplasia (6). Size and shape were extremely variable and not specific to lesion type. There was a tendency for pheochromocytomas (7), adenocarcinomas (5) and poorly differentiated blastoma (1) to be rounded masses. Adenomas (4), hyperplasia (7) and adrenal metastases (2) presented predominantly as nodules. No specificity in echogenicity was noted. Mineralization and bilaterality were present in both benign and malignant lesions. Vascular extension or the presence of a thrombus were suggestive but not specific signs of malignancy. Based on our preliminary study, ultrasonography is an effective method for localizing adrenal lesions and is helpful in assessing their extension. However, no definitive differentiation between benign and malignant lesions was possible using ultrasonographic criteria alone. PMID- 9402713 TI - Peripheral neuroblastoma in a dog. AB - Primitive neuroectodermal tumors are composed of primitive neuroepithelial cells and include tumors of the central and peripheral nervous system. Neuroblastoma, medulloblastoma and retinoblastoma are examples of these rare malignant tumors that usually occur in young patients. This report describes a peripheral neuroblastoma in a 2 year old Boxer that presented with signs of renal disease and a palpable abdominal mass. The purpose of this paper is to describe the clinical presentation, imaging and immunohistological studies of this abdominal tumor in a young dog and to review the literature. PMID- 9402714 TI - High-resolution parathyroid sonography. AB - The purpose of this study was to assess the diagnostic utility of parathyroid ultrasonography to differentiate causes of hypercalcemia in dogs. We analyzed qualitative and quantitative ultrasound imaging findings and clinical pathology data from 33 dogs that underwent parathyroid ultrasound examination as part of the diagnostic evaluation for hypercalcemia. Diagnoses of the diseases causing hypercalcemia included parathyroid carcinoma (n = 5), parathyroid adenoma (n = 15), parathyroid adenomatous hyperplasia (n = 6), chronic renal insufficiency (n = 3), and hypercalcemia of malignancy (n = 4). All parathyroid lesions were round or oval and hypoechoic compared with surrounding thyroid parenchyma. Adenomatous and adenocarcinomatous glands were 4 mm or larger in longest linear measurement on US examination and were statistically significantly larger than hyperplastic glands. (p < 0.001) Linear measurements of parathyroid glands acquired at the time of ultrasound examination correlated well with direct size determination after surgical excision. (r2 = 0.9, p < 0.0001) Parathyroid lesions > or = 4 mm are highly suspicious for parathyroid adenoma or carcinoma, while US lesions < 4 mm most likely represent primary adenomatous hyperplasia or secondary parathyroid hyperplasia. Parathyroid size estimation from ultrasound examination is an accurate predictor of true size. PMID- 9402715 TI - Ultrasound diagnosis: left ventricular thrombus in a cat with hypertrophic cardiomyopathy. PMID- 9402716 TI - The effect of intravenous diazepam on solid phase gastric emptying in normal cats. AB - Intravenous diazepam has been advocated as an appetite stimulant in anorexic cats. Diazepam has also been used to stimulate the intake of radiographic contrast medium-food mixture to determine the gastric emptying time of a solid meal. Diazepam has been suspected to delay gastric emptying in cats. One study found diazepam combined with valium to have little effect on gastric transit times in cats while diazepam alone accelerates gastric emptying in humans. The purpose of this study was to determine if diazepam influences gastric emptying times in normal cats. The gastric emptying half-time of solid food in normal, non diazepam treated cats has been previously determined using a scintigraphic technique using 99mTc-sulfur colloid to radiolabel solid dry food. The median gastric emptying half-time was 2.3 hours and the mean meal size was 16.1 grams. Gastric emptying half-times were determined in this study using diazepam as an appetite stimulant. The median gastric emptying half-times of diazepam treatment groups given both a 16.1 gram meal and a large meal were both significantly longer than the normal non-treated group (P < 0.05). Solid phase gastric emptying is therefore significantly delayed when diazepam is used as an appetite stimulant, irrespective of the volume of the meal. PMID- 9402718 TI - Subdural collection of contrast medium during cervical myelography. PMID- 9402717 TI - Slow release cisplatin combined with radiation for the treatment of canine nasal tumors. AB - Thirteen dogs with malignant tumors of the nasal cavity were treated with a combination of slow release cisplatin and megavoltage radiation. Radiation was delivered on a Monday through Friday schedule using a 6 MV linear accelerator. The median total dose was 49.5 Gy (range 49.5-56 Gy). Cisplatin was given using an open-cell polylactic acid polymer, impregnated with the drug and implanted intramuscularly at a distant site, as a slow release delivery system (OPLA-Pt [THM Biomedical, Inc]). The median dose used was 60 mg/m2 (range 60-100 mg/m2). When combined with radiation, this delivery system caused no systemic drug toxicity, and a local tissue reaction was seen in only two dogs. Acute side effects to normal tissue from radiation were not enhanced, as measured by subjective assessment. When compared to a group of historical controls that received radiation without OPLA-Pt, the dogs that received combined radiation and cisplatin had longer overall survival times, with a median of 580 days. The control group had a median survival of 325 days. Previously reported median survival times for comparable megavoltage radiation treatment range from 6 to 13 months. Some dogs in both groups also received adjubant chemotherapy but this did not influence survival time. By multivariate analysis, only the use of OPLA-Pt was found to significantly influence survival, with a p value of p = 0.023. Mega voltage radiation and slow release cisplatin appears to be a well tolerated combination that may favorably affect survival of dogs with nasal tumors. PMID- 9402719 TI - Comparison of histology with maternal and fetal serology for the diagnosis of abortion due to bovine neosporosis. AB - An indirect fluorescent antibody test was applied to sera from normally calving and aborting cows and to samples of pleural fluid from their aborted calves, and the antibody titres were compared with histology and immunocytochemistry for the diagnosis of Neospora-associated abortion. Two groups of aborting cows and a third group of cows which had calved normally were used; group A consisted of 36 cows which aborted calves showing characteristic non-suppurative inflammatory lesions in which Neospora was demonstrated by immunocytochemistry, group B consisted of 100 cows which aborted calves without histological evidence of neosporosis, and group C consisted of 128 normally calved cows which were sampled within one month of calving. Serology on the maternal sera and fetal fluids was highly specific and sensitive for Neospora infection although 5 per cent of the cows which aborted Neospora-negative calves and 4.7 per cent of the normally calved cows were also seropositive. Anti-Neospora antibodies were also detected in 7 per cent of the samples of fetal fluid from Neospora-negative abortions. PMID- 9402720 TI - Gross and ultrasonographic anatomy of the carpal flexor tendon sheath in horses. AB - This study was undertaken to establish the gross anatomy and the ultrasonographic appearance of the carpal digital flexor tendon sheath (carpal sheath) and the palmar carpal region in normal horses. The isolated forelimbs from 15 horses were used to study the morphology of the sheath and associated structures, including a detailed study of the location of the main blood vessels and nerves in that region. These limbs and the forelimbs of five live, sound horses were also examined ultrasonographically. The examination yielded good soft tissue detail of the tendons and ligaments, synovial and perisynovial tissues and larger blood vessels. There was a good correlation between the ultrasonographic and gross anatomical appearance of the limbs. The sheath cavity was only identified after it had been distended with water, and the various synovial recesses at the level of the carpal canal were poorly imaged. PMID- 9402721 TI - Canine dysautonomia resembling the Key-Gaskell syndrome in Germany. PMID- 9402722 TI - Appearance of acute PRRS-like symptoms in sow herds after vaccination with a modified live PRRS vaccine. PMID- 9402723 TI - Morantel resistance by Haemonchus placei in cattle. PMID- 9402724 TI - Beef exports: looking for an opening. AB - BSE, food safety and the veterinarian's role in certification were among matters discussed by the BVA's President, Mr Ted Chandler, at a speech in Ballymena on October 29. Speaking at the BVA's Northern Ireland dinner, at which representatives of Government, local veterinary associations, the farming community and other organisations were present, Mr Chandler emphasised the need for the Government to recognise the veterinary profession's vital contribution to food hygiene and its pivotal role in ensuring that standards are maintained. The main text of his speech is given here. PMID- 9402725 TI - Availability of medicines. PMID- 9402726 TI - Foxes and neosporosis. PMID- 9402727 TI - Salmonella typhimurium DT104 in the northeast USA. PMID- 9402728 TI - Equine foot care in an arid environment. PMID- 9402729 TI - Equine headshaking survey. PMID- 9402730 TI - Evolutionary dynamics of tandem repeats in the mitochondrial DNA control region of the minnow Cyprinella spiloptera. AB - Length variation due to tandem repeats is now recognized as a common feature of animal mitochondrial DNA; however, the evolutionary dynamics of repeated sequences are not well understood. Using phylogenetic analysis, predictions of three models of repeat evolution were tested for arrays of 260-bp repeats in the cyprinid fish Cyprinella spiloptera. Variation at different nucleotide positions in individual repeats supported different models of repeat evolution. One set of characters included several nucleotide variants found in all copies from a limited number of individuals, while the other set included an 8-bp deletion found in a limited number of copies in all individuals. The deletion and an associated nucleotide change appear to be the result of a deterministic, rather than stochastic, mutation process. Parallel origins of repeat arrays in different mitochondrial lineages, possibly coupled with a homogenization mechanism, best explain the distribution of nucleotide variation. PMID- 9402731 TI - The Bag320 satellite DNA family in Bacillus stick insects (Phasmatodea): different rates of molecular evolution of highly repetitive DNA in bisexual and parthenogenic taxa. AB - The Bag320 satellite DNA (satDNA) family was studied in seven populations of the stick insects Bacillus atticus (parthenogenetic, unisexual) and Bacillus grandii (bisexual). It was characterized as widespread in all zymoraces of B. atticus and in all subspecies of B. grandii. The copy number of this satellite is higher in the bisexual B. grandii (15%-20% of the genome) than in the parthenogenetic B. atticus (2%-5% of the genome). The nucleotide sequences of 12 Bag320 clones from B. atticus and 17 from B. grandii differed at 13 characteristic positions by fixed nucleotide substitutions. Thus, nucleotide sequences from both species cluster conspecifically in phylogenetic dendrograms. The nucleotide sequences derived from B. grandii grandii could be clearly discriminated from those of B. grandii benazzii and B. grandii maretimi on the basis of 25 variable sites, although all taxa come from Sicily. In contrast, the Bag320 sequences from B. atticus could not be discriminated accordingly, although they derive from geographically quite distant populations of its three zymoraces (the Italian and Greek B. atticus atticus, the Greek and Turkish B. atticus carius, and the Cyprian B. atticus cyprius). The different rate of evolutionary turnover of the Bag320 satDNA in both species can be related to their different modes of reproduction. This indicates that meiosis and chromosome segregation affect processes in satDNA diversification. PMID- 9402732 TI - Determination of the entire sequence of turtle CR1: the first open reading frame of the turtle CR1 element encodes a protein with a novel zinc finger motif. AB - CR1 elements are a family of retroposons. They are classified as long interspersed elements (LINEs) or non-long-terminal-repeat (non-LTR) retrotransposons, and they have been found in the genomes of many vertebrates. However, they have been only partially characterized, and only a 2-kb region of the 3' end of chicken CR1 has been sequenced. In the present study, we determined the entire consensus sequence of CR1 elements in the turtle genome, designated PsCR1. The first open reading frame (ORF1) of PsCR1 has two unusual arrangements of Cys residues. One of them includes a zinc finger motif, CX2CX14CX2C. The putative zinc finger has cysteine residues with identical spacing and a similar amino acid composition to those found in the species-specific transcription initiation factors SL1 and TIF-IB. The 5' untranslated region (5' UTR) of PsCR1 contains a sequence similar to part of the human L1 promoter, L1 site A, and several cis elements of the type found in eukaryotic genes. Within a region of about 500 bp, there are nine "E boxes," cis elements that are recognized by the basic helix-loop-helix (bHLH) family of proteins. This observation raises the possibility that cellular transcription factors that bind to these sequences might act in concert to regulate the expression of PsCR1. The extent of the sequence divergence of the 3' UTR of CR1 between species was found to be lower than the rate of nonsynonymous substitutions per site in ORF2, suggesting that a strict functional constraint must exist for this region. This result strongly suggests that the conserved 3'-end sequence of CR1 is the recognition site for the reverse transcriptase of CR1. A discussion is presented of a possible mechanism for the integration of CR1 elements and also of the intriguing possible recruitment of the reverse transcriptase for the retroposition of SINEs. PMID- 9402733 TI - A single lineage of r2 retrotransposable elements is an active, evolutionarily stable component of the Drosophila rDNA locus. AB - R2 elements are non-long-terminal-repeat (non-LTR) retrotransposons that insert specifically in the 28S rRNA genes of many insects. Previous reports concerning this element in the genus Drosophila have suggested that R2 elements are absent from many species of this genus, particularly those species from the subgenus Drosophila. In this report, we present an extensive study of the distribution and evolution of R2 elements in Drosophila. A PCR survey of 59 species from 23 species groups of the two major Drosophila subgenera found that R2 elements are present in all but two species of the melanogaster species subgroup. Phylogenetic analysis based on partial nucleotide sequences of R2 elements from 23 species demonstrates that the relationships of R2 elements are congruent with those of the Drosophila species phylogeny, suggesting that these elements have been vertically inherited since the divergence of this genus some 60 MYA. Sequence variation between different copies of R2 elements within each species was less than 0.16%, indicating that these elements are undergoing concerted evolution similar to that of the 28S genes. Several properties of the R2 sequences suggest that these elements depend on retrotransposition in addition to simple recombination to remain within the rDNA locus: the rates of synonymous substitutions averaged 4.8 times the rate of replacement substitutions, 82 of 83 R2 copies partially sequenced contained intact open reading frames, and, finally, length variation associated with the poly(A) 3' tails indicated that many R2 copies are the direct result of retrotransposition. PMID- 9402735 TI - Rates of DNA sequence evolution are not sex-biased in Drosophila melanogaster and D. simulans. AB - To determine whether male- or female-biased mutation rates have affected the molecular evolution of Drosophila melanogaster and D. simulans, we calculated the male-to-female ratio of germline cell divisions ([symbol: see text]) from germline generation data and the male-to-female ratio of mutation rate ([symbol: see text]) by comparing chromosomal levels of nucleotide divergence. We found that the ratio of germline cell divisions changes from indicating a weak female bias to indicating a weak male bias as the age of reproduction increases. The range of [symbol: see text] values that we observed, however, does not lead us to expect much, if any, difference in mutation rate between the sexes. Silent and intron nucleotide divergence were compared between nine loci on the X chromosome and nine loci on the second and third chromosomes. The average levels of nucleotide divergence were not significantly different across the chromosomes, although both silent and intron sites show a trend toward slightly more divergence on the X. These results indicate a lack of sex- or chromosome-biased molecular evolution in D. melanogaster and D. simulans. PMID- 9402734 TI - Phylogenetic analyses of the rbcL sequences from haptophytes and heterokont algae suggest their chloroplasts are unrelated. AB - Using the large subunit of RuBisCo (rbcL) sequences from cyanobacteria, proteobacteria, and diverse groups of algae and green plants, we evaluated the plastid relationship between haptophytes and heterokont algae. The rbcL sequences were determined from three taxa of heterokont algae (Bumilleriopsis filiformis, Pelagomonas calceolata, and Pseudopedinella elastica) and added to 25 published sequences to obtain a data set comprising 1,434 unambiguously aligned sites (approximately 98% of the total rbcL gene). Higher levels of mutational saturation in third codon positions were observed by plotting the pairwise substitutions with and without corrections for multiple substitutions at the same site for first and second codon positions only and for third positions only. In accordance with this finding phylogeny reconstructions were completed by omitting third codon positions, thus using 956 bp in weighted-parsimony and maximum likelihood analyses. The midpoint-rooted phylogenies showed two major clusters, one containing cyanobacteria, glaucocystophytes, a phototrophic euglenoid, chlorophytes, and embryophytes (the green lineage), the other containing proteobacteria, haptophytes, red algae, a cryptophyte, and heterokont algae (the non-green lineage). In the nongreen lineage, the haptophytes formed a sister group to the clade containing heterokont algae, red algae, and the cryptophyte Guillardia theta. This branching pattern was well supported in terms of bootstrap values in weighted-parsimony and maximum-likelihood analyses (100% and 92%, respectively). However, the phylogenetic relationship among red algae, heterokonts, and a cryptophyte taxon was not especially well resolved. A four cluster analysis was performed to further explore the statistical significance of the relationship between proteobacteria, red algae (including and excluding Guillardia theta), haptophytes, and heterokont algae. This test strongly favored the hypothesis that the heterokonts and red algae are more closely related to each other than either is to proteobacteria or haptophytes. Hence, this molecular study based on a plastid-encoded gene provides additional evidence for a distant relationship between haptophytes and the heterokont algae. It suggests an evolutionary scenario in which the ancestor of the haptophyte lineage engulfed a phototrophic eukaryote and, more recently, the heterokont lineage became phototrophic by engulfing a red alga. PMID- 9402736 TI - Multiregional introgressions inferred from the mitochondrial DNA phylogeny of a hybridizing species complex of gobiid fishes, genus Tridentiger. AB - Partial sequences of the cytochrome b gene (402 bp) in mtDNA were determined for brackishwater gobiid fishes, genus Tridentiger, collected from geographically distant locations in the Japanese Archipelago, and their interspecific and geographic variations were analyzed and compared. Contrary to the results of a previous allozyme analysis which revealed the existence of considerable genetic divergence (Nei's genetic distance > 0.5) between T. obscurus and T. brevispinis, the mtDNA haplotypes (mitotypes) of these two species were very similar and could not be distinguished by any of the neighbor-joining, maximum-likelihood or parsimony analyses. Hybrid individuals between the two species were also found, with several mitotypes being shared by both species and their hybrids. The phylogenetic relationships of mitotypes were divided into three subgroups, the geographical distributions of the latter being allied to geographical features of the Archipelago. These results suggested the occurrence of multiregional introgression between the two species, with mitotypes transferring from one species to the other. PMID- 9402737 TI - The complete mitochondrial genome of Alligator mississippiensis and the separation between recent archosauria (birds and crocodiles). AB - The complete mitochondrial genome of the alligator, Alligator mississippiensis, was sequenced. The size of the molecule is 16,642 nucleotides. Previously reported rearrangements of tRNAs in crocodile mitochondrial genomes were confirmed and, relative to mammals, no other deviations of gene order were observed. The analysis of protein-coding genes of the alligator showed an evolutionary rate that is roughly the same as in mammals. Thus, the evolutionary rate in the alligator is faster than that in birds as well as that in cold blooded vertebrates. This contradicts hypotheses of constant body temperatures or high metabolic rate being correlated with elevated molecular evolutionary rates. It is commonly acknowledged that birds are the closest living relatives to crocodiles. Birds and crocodiles represent the only archosaurian survivors of the mass extinction at the Cretaceous/Tertiary boundary. On the basis of mitochondrial protein-coding genes, the Haemothermia hypothesis, which defines birds and mammals as sister groups and thus challenges the traditional view, could be rejected. Maximum-likelihood branch length data of amino acid sequences suggest that the divergence between the avian and crocodilian lineages took place at approximately equal to 254 MYA. PMID- 9402738 TI - Lactate dehydrogenase (LDH) gene duplication during chordate evolution: the cDNA sequence of the LDH of the tunicate Styela plicata. AB - L-Lactate dehydrogenase (L-LDH, E.C. 1.1.1.27) is encoded by two or three loci in all vertebrates examined, with the exception of lampreys, which have a single LDH locus. Biochemical characterizations of LDH proteins have suggested that a gene duplication early in vertebrate evolution gave rise to Ldh-A and Ldh-B and that an additional locus, Ldh-C arose in a number of lineages more recently. Although some phylogenetic studies of LDH protein sequences have supported this pattern of gene duplication, others have contradicted it. In particular, a number of studies have suggested that Ldh-C represents the earliest divergence among vertebrate LDHs and that it may have diverged from the other loci well before the origin of vertebrates. Such hypotheses make explicit statements about the relationship of vertebrate and invertebrate LDHs, but to date, no closely related invertebrate LDH sequences have been available for comparison. We have attempted to provide further data on the timing of gene duplications leading to multiple vertebrate LDHs by determining the cDNA sequence of the LDH of the tunicate Styela plicata. Phylogenetic analyses of this and other LDH sequences provide strong support for the duplications giving rise to multiple vertebrate LDHs having occurred after vertebrates diverged from tunicates. The timing of these LDH duplications is consistent with data from a number of other gene families suggesting widespread gene duplication near the origin of vertebrates. With respect to the relationships among vertebrate LDHs, our data are not consistent with previous claims that Ldh-C represented the earliest divergence. However, the precise relationships among some of the main lineages of vertebrate LDHs were not resolved in our analyses. PMID- 9402739 TI - Intragenic duplication and divergence in the spectrin superfamily of proteins. AB - Many structural, signaling, and adhesion molecules contain tandemly repeated amino acid motifs. The alpha-actinin/spectrin/dystrophin superfamily of F-actin crosslinking proteins contains an array of triple alpha-helical motifs (spectrin repeats). We present here the complete sequence of the novel beta-spectrin isoform beta(Heavy)-spectrin (beta H). The sequence of beta H supports the origin of alpha- and beta-spectrins from a common ancestor, and we present a novel model for the origin of the spectrins from a homodimeric actin-crosslinking precursor. The pattern of similarity between the spectrin repeat units indicates that they have evolved by a series of nested, nonuniform duplications. Furthermore, the spectrins and dystrophins clearly have common ancestry, yet the repeat unit is of a different length in each family. Together, these observations suggest a dynamic period of increase in repeat number accompanied by homogenization within each array by concerted evolution. However, today, there is greater similarity of homologous repeats between species than there is across repeats within species, suggesting that concerted evolution ceased some time before the arthropod/vertebrate split. We propose a two-phase model for the evolution of the spectrin repeat arrays in which an initial phase of concerted evolution is subsequently retarded as each new protein becomes constrained to a specific length and the repeats diverge at the DNA level. This evolutionary model has general applicability to the origins of the many other proteins that have tandemly repeated motifs. PMID- 9402740 TI - Nucleotide polymorphism in the acidic chitinase locus (ChiA) region of the wild plant Arabidopsis thaliana. AB - To investigate DNA variation in natural plant populations, a 1.8-kb region of the acidic chitinase locus (ChiA)was analyzed for 17 ecotypes of Arabidopsis thaliana sampled worldwide and 3 Arabis species in Japan. As in the Adh region, dimorphism was detected throughout the investigated ChiA region, suggesting the possibility that dimorphic DNA variation exists in the entire nuclear genome of A. thaliana. The ChiA region was divided into two blocks by an intragenic recombination between two parental sequence types, which diverged 7.4 MYA under the assumption that nucleotide mutation rate per site per year is mu = 10(-9). Nucleotide diversity in the entire ChiA region was 0.0104. Tajima's test was significantly negative for both nucleotide and indel variations, which was manifested as an excess of unique polymorphisms. However, the level and pattern of polymorphism in the ChiA region were inconsistent with simple theoretical explanations. The HKA test detected no difference in the levels of intra- and interspecific variations between the ChiA and Adh regions. In the ChiA coding region, no difference in the patterns of synonymous and replacement variation was found in intra- and interspecific comparisons by the MK test. Although it was difficult to determine the exact genetic mechanism acting on the ChA locus, these results suggested that the ChA locus region was under the same genetic mechanism before and after the establishment of A. thaliana as a species. PMID- 9402741 TI - Characterization and molecular analysis of Adh retrosequences in species of the Drosophila obscura group. AB - Retrosequences, genes, and pseudogenes originated by retrotranscription are frequent components of vertebrate genomes, but they have only occasionally been described in invertebrates. In Drosophila, very few retrosequences have been reported, among them those of alcohol dehydrogenase (Adh) and phosphoglyceromutase (Pglym). Although 52 Adh gene sequences are available for comparison, Adh retrosequences have been described only in the sibling species D. teissieri and D. yakuba (melanogaster subgroup) and in D. subobscura (obscura subgroup). Here, we report the presence of Adh retrosequences in two closely related species of D. subobscura: D. madeirensis and D. guanche. Extensive sequence comparisons with their functional paralogs suggest separate retrotranscriptional events: one in the melanogaster subgroup in the ancestor of D. teissieri and D. yakuba, and the other in the obscura subgroup before the radiation of the lineages leading to D. subobscura, D. madeirensis, and D. guanche. In the former, the Adh retrotranscript originated a new expressed gene, named jingwei. However, in the obscura Adh retrosequences, retention of codon bias and higher Ks than Ka values, both distinctive evolutionary features supporting functionality, have to be considered together with a frameshift, premature stop codons, and other nucleotide substitutions, which, added to the lack of the original promoter elements, suggest that they are pseudogenes. At least two different Adh retrosequences have been characterized in each of the obscura species, and their phylogenetic analysis indicates that paralogs and their flanking genomic regions share a higher degree of similarity than orthologous sequences. Two alternative hypotheses could explain this current organization and structure: either a multiplication event occurred independently in each species, or gene conversion events should be invoked after a single duplication in the species ancestor. The significance of retrotranscriptional events in the evolution of invertebrate genomes is discussed. PMID- 9402743 TI - Small-sample tests of episodic adaptive evolution: a case study of primate lysozymes. PMID- 9402742 TI - Early evolution of metazoan serine/threonine and tyrosine kinases: identification of selected kinases in marine sponges. AB - The phylum Porifera (sponges) was the first to diverge from the common ancestor of the Metazoa. In this study, six cDNAs coding for protein-serine/threonine kinases (PS/TKs) are presented; they have been isolated from libraries obtained from the demosponges Geodia cydonium and Suberites domuncula and from the calcareous sponge Sycon raphanus. Sequence alignments of the catalytic domains revealed that two major families of PS/TK, the "conventional" (Ca(2+)-dependent) protein kinase C (PKC), the cPKC subfamily, as well as the "novel" (Ca(2+) independent) PKC (nPKC), form two separate clusters. In each cluster, the sequence from S. raphanus diverges first. To approach the question about the origin of protein-tyrosine kinases (PTK), which are found only in Metazoa, we analyzed two additional PS/TKs which have been cloned from S. domuncula: the stress-responsive protein kinase (KRSvSD) and the protein-kinase-C-related kinase (PRKvSD). The construction of the phylogenetic tree, comprising the eight PS/TKs and the PTK cloned previously from G. cydonium, revealed that the PTK derived from the branch including the KRSvSD kinase. These data facilitate the first molecular approach to elucidate the origin of metazoan PTK within the PS/TK superfamily. PMID- 9402744 TI - Molecular evolution of the amy multigenes in the subgenus Sophophora of Drosophila. PMID- 9402745 TI - Pharmacology of chronic oral daily administration of idarubicin. AB - Idarubicin (4-demethoxydaunorubicin) (IDA) is a daunorubicin analogue with substantial activity in hematologic malignancies and solid tumors. Among several reasons, IDA is of interest because of its main metabolite derivative, the C-13 alcohol analogue, idarubicinol (IDOL). Previous studies have suggested that IDOL, unlike other anthracycline metabolic derivatives, possesses a striking growth inhibitory activity in tumor cell lines. This suggests that IDOL, like IDA could be useful in circumventing MDR. IDA is bioavailable in an oral dosage form. After oral administration of IDA to the patients, the concentration of IDOL quickly exceeds that of IDA and is retained in the plasma for a longer period. Hence, administration of IDA to cancer patients results ina much greater overall exposure of the tumor to IDOL than to the parent compound. At the Oncology Center (CRO) in Aviano we performed a dose-finding and pharmacokinetic (PK) study of chronic daily oral IDA with intrapatient escalation in patients with metastatic breast cancer (MBC). All the patients were pretreated with anthracyclines (the cumulative dose was 530 mg and 264 mg, respectively, for epirubicin and (DOX) and had at admittance a PS < or = 2 and a left ventricular ejection fraction > 50%. IDA (1 mg capsules) was administered orally twice a day for 21 days every two weeks. Treatment was continued at escalating doses until progression or intolerance. Twenty-five patients were enrolled. MTD has not yet been reached and clinical results are reported in Table 1. Treatment was well-tolerated in all but one patient (300 ANC at day 28). Three patients had tox G3 ANC for more than three weeks after 3, 6, and 7 mg doses, respectively. Two of them stopped chemotherapy after 1 cycle and 1 patient stopped after 2 cycles (6 mg doses). Despite previous treatments with anthracyclines (the mean cumulative dose before entering the study was 530 and 264 mg, respectively, for epirubicin and DOX) no cardiotoxicity due to IDA treatment was observed. This trial demonstrates the feasibility of chronic daily IDA administration. At the dosage reported, treatment was generally well tolerated. The PK findings (high IDOL concentrations) and the unexpected G4 myelotoxicity in patients with the highest IDOL plasma concentrations suggest that IDOL is clinically relevant. PMID- 9402746 TI - Low-dose long-term oral idarubicin in maintenance treatment of elderly acute myeloid leukemia. AB - BACKGROUND AND OBJECTIVE: Low-dose long-term oral IDA may play a role in maintainance treatment of elderly patients with AML; in fact, continuous exposure to IDA and IDAol could be efficacious in the disease control possibly inducing cell-differentiation and/or apoptosis. METHODS: We enrolled 25 previous responder patients in standard induction therapy to receive maintenance oral IDA 5 mg daily on days 1-14 at 2-week intervals for at least 6 months. We also evaluated the cell-cycle and apoptosis in leukemic cells from patients after IDA administration and, as a control, from HL60 lines exposed to IDA and IDAol in vitro. RESULTS: Long-term long-dose IDA was well-tolerated. Neutrophil and platelet count never below under 1 x 10(9)/L and 50 x 10(9)/L respectively in CR patients, and no infectious complications were encountered. Non-hematological toxicity was also acceptable: easily controlled nausea and vomiting, non-recorded diarrhea or mucositis were reported. The convenience of oral administration contributed to excellent compliance. DNA analysis performed in vivo after IDA and IDAol exposure showed an increase of G2/M cell frequencies and evidence of sub-G1 peak. INTERPRETATION AND CONCLUSIONS: In conclusion, long-term low doses of oral IDA would appear valuable as a maintenance regimen for elderly patients. Our results seem to confirm the preliminary hypothesis that IDA + IDAol induce an increase of apoptosis in leukemic cells. PMID- 9402747 TI - Idarubicin and cytosine arabinoside in the induction and maintenance therapy of high-risk myelodysplastic syndromes. AB - BACKGROUND AND OBJECTIVE: Recently, the results of a few pilot studies have shown the efficacy of the association of idarubicin (IDA) and cytosine arabinoside (Ara C), already successfully employed in acute myeloid leukemia (AML), for remission induction in patients with myelodysplastic syndrome (MDS). We set out to evaluate in a multicenter study the efficacy and tolerability of an intensive therapy with IDA and Ara-C in patients with RAEB and RAEB-t, the rate and duration of CR and the overall survival in adults treated with full doses and in the elderly treated with lower doses; furthermore, we investigated the efficacy of low-dose maintenance chemotherapy. METHODS: Pretreated adult patients with de novo RAEB and RAEB-t, meeting at least one of the following criteria, were included: neutrophils < 0.5 x 10(9)/L or moderate neutropenia with infectious episodes, platelets < 30 x 10(9)/L or moderate thrombocytopenia but with bleeding symptoms, transfusion > 4 red cell units/months, rapid increase of bone marrow blasts. Induction treatment consisted of a cycle with IDA and Ara-C. Adult patients less than 65 years old were treated with the following doses: Ara-C 1 g/m2/day i.v. 6 hour infusion, on days 1-4, IDA 10 mg/m2/day i.v., on days 1-3. Elderly patients (> or = 65 yrs) were treated with lower doses: Ara-C 1 g/m2/day 6 hours infusion, on days 1 and 2, IDA 10 mg/m2/day i.v., on days 1 and 2. Responders followed a consolidation course identical to induction. RESULTS: From February 1994 to February 1997, 25 patients were enrolled, 20 males and 5 females aged between 22 and 76, 10 were > or = 65 years old, 7 had RAEB and 18 had RAEB-t. Twelve cases (48%) achieved complete remission (CR), 7 cases (28%) achieved partial remission, 4 patients were resistant and two patients (8%) died during the aplastic phase. A significantly higher CR rate was found in younger patients (p = 0.036), while gender, FAB subtype, presence of Auer rods, cytogenetic findings, and the interval from diagnosis to treatment did not significantly influence CR achievement. INTERPRETATION AND CONCLUSIONS: Our results show that in de novo RAEB and RAEB-t, the employed treatment with IDA and Ara-C is associated with satisfactory frequency of response with acceptable toxicity. PMID- 9402748 TI - Idarubicin in induction treatment of acute myeloid leukemia in the elderly. AB - BACKGROUND AND OBJECTIVE: The best approach to treatment of acute myeloid leukemia (AML) in elderly patients remains controversial. Intensive chemotherapy is the treatment of choice in selected patients, but age related changes might affect the pharmacokinetics of antineoplastic agents resulting in enhanced toxicity. We report our experience in elderly patients treated with idarubicin at attenuated doses plus cytarabine and etoposide. METHODS: Sixty-six AML patients, median age 66, with progressive disease and high tumor burden received idarubicin 8 mg/sqm i.v. d 1,3,5; cytarabine 200 mg/sqm by continuous i.v. infusion d 1-7; etoposide 60 mg/sqm i.v. d 1-5. A second course with the same drugs was planned irrespective of complete remission (CR) achievement. No consolidation was given; 44% had a documented preexisting myelodysplasia, 45% had a documented preexisting myelodysplasia, 45% presented with fever. Promyelocytic leukemias were excluded. RESULTS: Thirty-five patients (53%) achieved CR and 9 PR for an overall response rate of 67%. Nine of them (13%) died early or during the aplastic phase. Preexisting myelodysplasia had no significant impact on CR achievement. Resistant disease was associated with CD7 phenotype and unfavorable karyotype. Overall survival and disease free survival were 14 and 13 months, respectively. The major toxicity consisted of infectious complications (WHO > 2 in 24% of patients). Six patients died for infection, 2 for heart failure, 1 for pulmonary embolism. INTERPRETATION AND CONCLUSIONS: This induction regimen with attenuated doses of idarubicin is feasible and effective, but long-term survival remains an unresolved problem. Alternative post remission approaches are advisable in the aim of improving the remission duration. PMID- 9402749 TI - A single high dose of idarubicin combined with high-dose ABA-C (MSKCC ALL-3 protocol) in adult and pediatric patients with acute lymphoblastic leukemia. Experience at the University La Sapienza of Rome. AB - BACKGROUND AND OBJECTIVE: The anthracycline analogue idarubicin, either alone or in combination with other antineoplastic drugs, has shown antileukemic activity in relapsed and refractory acute lymphoblastic leukemia (ALL). In an attempt to minimize the non-hematologic toxicity and obtain a potent antileukemic effect, MSKCC activated a pilot study in previously treated adult ALL, using HD-ARA-C combined with idarubicin administered as a single high-dose infusion. We herein report our experience with a series of pediatric and adult high risk ALL and NHL patients treated with the protocol above, which confirms its feasibility, response rate and individual compliance. METHODS: In a clinical phase II study the combination of a single high dose (HD) idarubicin and HD cytosine-arabinoside (ARA-C), as designed at the Memorial Sloan Kettering Cancer Center, was applied to 70 adults and children with refractory or early relapse acute lymphoblastic leukemia (ALL) and T-cell lymphoblastic non-Hodgkin's lymphoma (NHL). Therapy consisted of HD-ARA-C 3 g/m2/d on days 1-5, idarubicin 40 mg/m2 on day 3, prophylactic intrathecal methotrexate on days 1 and 4, and G-CSF 5 mg/kg/d s.c. from day 7 to hematopoietic reconstitution (PMN > 0.5 x 10(9)/L). RESULTS: Fifty five of the 70 patients (78%) achieved complete remission (CR), four died in aplasia due to infection and 11 were non-responders. Recovery of blood counts occurred at a median of 21 days from the start of treatment. Non-hematologic side effects were extremely limited and consisted predominantly of infections. INTERPRETATION AND CONCLUSIONS: In view of the highly unfavorable series of patients selected, this study confirms the feasibility and antileukemic activity of the HD-idarubicin + HD- ARA-C combination in patients with refractory and early relapse ALL and NHL. The excellent tolerance to this regimen does not preclude bone marrow transplantation as post-remission treatment. PMID- 9402750 TI - Idarubicin in low-grade non-Hodgkin's lymphomas. AB - The majority of responses produced in patients with low-grade lymphomas are unique among non-Hodgkin's lymphomas (NHL), and even with a more intensive chemotherapy regimen, they are only partial; the very few complete responses which are induced are usually of short duration and do not influence overall survival. There is, therefore, a need for new approaches to the management of low grade NHLs. Studies are currently in progress to assess the potential benefits in the treatment of NHL offered by new drugs, including fludarabine, idarubicin and 2-chlorodeoxyadenosine. In order to evaluate idarubicin in combination with purine analogs, we used a combination of fludarabine and idarubicin, called the FLU-ID regimen, to treat 10 patients with recurrent low-grade NHL. Of the 10 patients, 2 (20%) achieved complete response, 5 (50%) partial response, and the remaining 3 showed no benefit from the treatment. The 2 CR patients are still in remission after 12 and 14 months, respectively. The median duration of overall survival of all patients was 18 months. These results indicate the efficacy of the FLU-ID regimen in inducing a good remission rate with moderate side effects in recurrent low-grade NHL. On the basis of this pilot study, we planned a cooperative randomized trial for untreated patients. PMID- 9402751 TI - [Paclitaxel (Taxol) opens new perspectives on bronchial carcinoma. 8th World Conference on Lung Cancer. Dublin, 10-15 August 1997]. PMID- 9402752 TI - Identification and characterization of a catalytic base in bacterial luciferase by chemical rescue of a dark mutant. AB - Mutation of the His44 residue of the alpha subunit of Vibrio harveyi luciferase to an alanine was known to reduce the enzyme bioluminescence activity by five orders of magnitude [Xin, X., Xi, L., and Tu, S.-C. (1991) Biochemistry 30, 11255 11262]. We found that the residual activity of the alpha H44A luciferase was markedly enhanced by exogenously added imidazole and other simple amines. The peak luminescence intensity in nonturnover assays was linearly proportional to levels of alpha H44A and the rescue agent, indicating a lack of significant binding under our experimental conditions. The rescue effect of imidazole was pH dependent and quantitatively correlated well with the amount of imidazole base. The rescue efficiencies of imidazole and amines were found to be regulated by both their molecular volume and pKa. A Bronsted analysis revealed a beta value of 0.8 +/- 0.1. The enhancement of alpha H44A activity by imidazole took place after the formation of the flavin 4a-hydroperoxide intermediate. The predominant form of the flavin 4a-hydroperoxide intermediate generated by alpha H44A was inactive in bioluminescence, but was reactive with the aldehyde substrate for bioluminescence in the presence of imidazole. These findings, taken together, provide evidence for assigning a role for the alpha His44 imidazole as a catalytic base in the luciferase reaction. This study provides the first characterization of a catalytic residue for bacterial luciferase and the first demonstration of the rescue of a histidine-mutated enzyme by exogenous imidazole and amines. PMID- 9402753 TI - Decreased stability of transforming growth factor beta type II receptor mRNA in RER+ human colon carcinoma cells. AB - Transforming growth factor beta (TGF-beta) is a potent inhibitor of cell growth and tumor progression. Previous work has shown that loss of functional TGF-beta type II receptor (RII) due to a frameshift mutation in the 5' half of the RII gene leads to TGF-beta resistance in a highly progressed, RER+ human colon carcinoma cell line designated HCT116. Expression of this mutated RII gene was highly repressed in RER+ cell lines such as HCT116 and RKO, as analyzed by RNase protection assays. Nuclear run-on and RII promoter-reporter (CAT) assays showed that the transcriptional levels of the RII gene in these RER+ cells were not reduced, compared to RII-expressing cells. However, the half-lives of the RII mRNA, as analyzed by RNase protection assays following actinomycin D treatment, were significantly decreased. This suggested that the decreased expression of the RII gene mutant was due to decreased mRNA stability. Furthermore, RII mRNA from HCT116 transfected with wild-type RII had a longer half-life than the endogenous mutated RII mRNA. A dominant negative RII mutant, which encodes a similarly truncated RII protein as HCT116 but lacks the extensive 3' untranslated region of RII mRNA, gave the same half-life as endogenous wild-type RII mRNA. We conclude that the frameshift mutation which results in a premature stop codon in the 5' half of the mRNA transcript accounts for the reduced RII mRNA levels in RER+ cells. PMID- 9402754 TI - Expression, purification, and inhibitory properties of human proteinase inhibitor. AB - In a previous report, the cDNA for human proteinase inhibitor 8 (PI8) was first identified, isolated, and subcloned into a mammalian expression vector and expressed in baby hamster kidney cells. Initial studies indicated that PI8 was able to inhibit the amidolytic activity of trypsin and form an SDS-stable approximately 67-kDa complex with human thrombin [Sprecher, C. A., et al. (1995) J. Biol Chem. 270, 29854-29861]. In the present study, we have expressed recombinant PI8 in the methylotropic yeast Pichia pastoris, purified the inhibitor to homogeneity, and investigated its ability to inhibit a variety of proteinases. PI8 inhibited the amidolytic activities of porcine trypsin, human thrombin, human coagulation factor Xa, and the Bacillus subtilis dibasic endoproteinase subtilisin A through different mechanisms but failed to inhibit the Staphylococcus aureus endoproteinase Glu-C. PI8 inhibited trypsin in a purely competitive manner, with an equilibrium inhibition constant (Ki) of less than 3.8 nM. The interaction between PI8 and thrombin occurred with a second-order association rate constant (kassoc) of 1.0 x 10(5) M-1 s-1 and a Ki of 350 pM. A slow-binding kinetics approach was used to determine the kinetic constants for the interactions of PI8 with factor Xa and subtilisin A. PI8 inhibited factor Xa via a two-step mechanism with a kassoc of 7.5 x 10(4) M-1 s-1 and an overall Ki of 272 pM. PI8 was a potent inhibitor of subtilisin A via a single-step mechanism with a kassoc of 1.16 x 10(6) M-1 s-1 and an overall Ki of 8.4 pM. The interaction between PI8 and subtilisin A may be of physiological significance, since subtilisin A is an evolutionary precursor to the intracellular mammalian dibasic processing endoproteinases. PMID- 9402755 TI - Mechanism of formation of novel covalent drug.DNA interstrand cross-links and monoadducts by enediyne antitumor antibiotics. AB - The potent enediyne antitumor antibiotic C1027 has been previously reported to induce novel DNA interstrand cross-links and drug monoadducts under anaerobic conditions [Xu et al. (1997) J. Am. Chem. Soc. 119, 1133-1134]. In the present study, we explored the mechanism of formation of these anaerobic DNA lesions. We found that, similar to the aerobic reaction, the diradical species of the activated drug initiates anaerobic DNA damage by abstracting hydrogen atoms from the C4', C1', and C5' positions of the A1, A2, and A3 nucleotides, respectively, in the most preferred 5'GTTA1T/5'ATA2A3C binding sequence. It is proposed that the newly generated deoxyribosyl radicals, which cannot undergo oxidation, likely add back onto the nearby unsaturated ring system of the postactivated enediyne core, inducing the formation of interstrand cross-links, connecting either A1 to A2 or A1 to A3, or drug monoadducts mainly on A2 or A3. Comparative studies with other enediynes, such as neocarzinostatin and calicheamicin gamma 1I under similar reaction conditions indicate that the anaerobic reaction process is a kinetically competitive one, depending on the proximity of the drug unsaturated ring system or dioxygen to the sugar radicals and their quenching by other hydrogen sources such as solvent or thiols. It was found that C1027 mainly generates interstrand cross-links, whereas most of the anaerobic lesions produced by neocarzinostatin are drug monoadducts. Calicheamicin gamma 1 (1) was found to be less efficient in producing both lesions. The anaerobic DNA lesions induced by enediyne antitumor antibiotics may have important implications for their potent cytotoxicity in the central regions of large tumors, where relative anaerobic conditions prevail. PMID- 9402756 TI - Regional trauma systems. PMID- 9402757 TI - Emergency medical admissions: taking stock and planning for winter. PMID- 9402758 TI - Commissioning specialist services in the NHS. PMID- 9402759 TI - Lumbar puncture needn't be a headache. PMID- 9402760 TI - UK government fails its first test on public health. PMID- 9402762 TI - Clearing house needed for specialist treatments. PMID- 9402761 TI - Climate change: decision time in Kyoto. PMID- 9402763 TI - Medical decisions must be logically defensible. PMID- 9402765 TI - BMA wants licensing of cannabis to be changed. PMID- 9402766 TI - US journal finds lower risk from diet pills. PMID- 9402767 TI - Scottish trusts miss cataract surgery targets. PMID- 9402768 TI - US hospital mergers threaten reproductive services. PMID- 9402769 TI - UK needs more cancer specialists. PMID- 9402770 TI - UK government consults on clinical disputes. PMID- 9402771 TI - GPs will be at heart of overhaul of NHS. PMID- 9402772 TI - Neonatal screening recommended for hearing impairment. PMID- 9402773 TI - Association of upper gastrointestinal toxicity of non-steroidal anti-inflammatory drugs with continued exposure: cohort study. AB - OBJECTIVES: To determine the profile of risk of upper gastrointestinal toxicity during continuous treatment with, and after cessation of, non-steroidal anti inflammatory drugs. DESIGN: Cohort study with a prospectively constructed, population based, record linkage database containing details of exposure to all community dispensed non-steroidal anti-inflammatory drugs and also all admissions to hospital for upper gastrointestinal diagnoses. SETTING: The population of Tayside, Scotland. SUBJECTS: 52,293 subjects aged 50 and over who received one or more non-steroidal anti-inflammatory between 1 January 1989 and 31 December 1991 and 73,792 subjects who did not receive one during the same period (controls). MAIN OUTCOME MEASURES: Admission to hospital for upper gastrointestinal bleeding and perforation, and admission for other upper gastrointestinal diagnoses. RESULTS: About 2% of the non-steroidal anti-inflammatory cohort were admitted with an upper gastrointestinal event during the study period compared with 1.4% of controls. The risk of admission for upper gastrointestinal haemorrhage and perforation was constant during continuous non-steroidal anti-inflammatory exposure and carried over after the end of exposure. The results were similar for admissions for all upper gastrointestinal events. CONCLUSION: This study provides evidence that non-steroidal anti-inflammatory toxicity persists with continuous exposure. There seems to be carryover toxicity after the end of prescribing. These findings have implications for the management of patients requiring non steroidal anti-inflammatory drugs. PMID- 9402774 TI - Randomised trial of octreotide for long term management of cirrhosis after variceal haemorrhage. AB - OBJECTIVE: To assess the efficacy of long term octreotide as adjuvant treatment to programmed endoscopic sclerotherapy after acute variceal haemorrhage in cirrhotic portal hypertension. DESIGN: Randomised clinical trial. SETTING: University hospital. SUBJECTS: 32 patients with cirrhotic portal hypertension. INTERVENTIONS: Programmed injection sclerotherapy with subcutaneous octreotide 50 micrograms twice daily for 6 months, or programmed injection sclerotherapy alone. MAIN OUTCOME MEASURES: Episodes of recurrent variceal bleeding and survival. RESULTS: Significantly fewer patients receiving combined octreotide and sclerotherapy had episodes of recurrent variceal bleeding compared with patients given sclerotherapy alone (1/16 v 7/16; P = 0.037, Fisher's exact test), and their survival was significantly improved (P < 0.02, log rank test); this improvement was maintained for 12 months after the end of the study. Combined treatment also resulted in a sustained decrease in portal pressure (median decrease -6.0 mm Hg, interquartile range -10 to -4.75 mm Hg, P = 0.0002) compared with sclerotherapy alone (median increase 1.5 mm Hg, interquartile range 0.25 to 3.25 mm Hg), as well as a significant improvement in liver function as assessed by plasma concentrations of bilirubin, albumin, and alanine aminotransferase and by hepatocyte metabolism of aminopyrine labelled with carbon-14. CONCLUSION: Long term octreotide may be a valuable adjuvant to endoscopic sclerotherapy for acute variceal haemorrhage in cirrhotic portal hypertension. PMID- 9402775 TI - Does malnutrition in utero determine diabetes and coronary heart disease in adulthood? Results from the Leningrad siege study, a cross sectional study. AB - OBJECTIVE: To investigate the relation between decreased maternal food intake and risk factors for coronary heart disease in adult life. DESIGN: Cross sectional study. SUBJECTS: 169 subjects exposed to malnutrition in utero (intrauterine group) during the siege of Leningrad (now St Petersburg) in 1941-4; 192 subjects born in Leningrad just before rationing began, before the siege (infant group); and 188 subjects born concurrently with the first two groups but outside the area of the siege (unexposed group). SETTING: Ott Institute of Obstetrics and Gynaecology, St Petersburg. MAIN OUTCOME MEASURES: Development of risk factors for coronary heart disease and diabetes mellitus-obesity, blood pressure, glucose tolerance, insulin concentrations, lipids, albumin excretion rate, and clotting factors. RESULTS: There was no difference between the subjects exposed to starvation in utero and those starved during infant life in: (a) glucose tolerance (mean fasting glucose: intrauterine group 5.2 (95% confidence interval 5.1 to 5.3), infant group 5.3 (5.1 to 5.5), P = 0.94; mean 2 hour glucose: intrauterine group 6.1 (5.8 to 6.4), infant group 6.0 (5.7 to 6.3), P = 0.99); (b) insulin concentration; (c) blood pressure; (d) lipid concentration; or (e) coagulation factors. Concentrations of von Willebrand factor were raised in the intrauterine group (156.5 (79.1 to 309.5)) compared with the infant group (127.6 (63.9 to 254.8); P < 0.001), and female subjects in the intrauterine group had a stronger interaction between obesity and both systolic (P = 0.01) and diastolic (P = 0.04) blood pressure than in the infant group. Short adult stature was associated with raised concentrations of glucose and insulin 2 hours after a glucose load-independently of siege exposure. Subjects in the unexposed group had non-systematic differences in subscapular to triceps skinfold ratio, diastolic blood pressure, and clotting factors compared with the exposed groups. CONCLUSIONS: Intrauterine malnutrition was not associated with glucose intolerance, dyslipidaemia, hypertension, or cardiovascular disease in adulthood. Subjects exposed to malnutrition showed evidence of endothelial dysfunction and a stronger influence of obesity on blood pressure. PMID- 9402776 TI - Commentary: a hypothesis challenged. PMID- 9402777 TI - Effectiveness of a regional trauma system in reducing mortality from major trauma: before and after study. AB - OBJECTIVE: To assess the effect of the development of an experimental trauma centre and regional trauma system on the survival of patients with major trauma. DESIGN: Controlled before and after study examining outcomes between 1990 and 1993, spanning the introduction of the system in 1991-2. SETTING: Trauma centre in North Staffordshire Royal Infirmary and five associated district general hospitals in the North West Midlands regional trauma system, and two control regions in Lancashire and Humberside. SUBJECTS: All trauma patients taken by the ambulance services serving the regions or arriving other than by ambulance with injury severity scores > 15, whether or not they had vital signs on arrival at hospital. MAIN OUTCOME MEASURES: Survival rates standardised for age, severity of injury, and revised trauma score. RESULTS: In 1990, 33% of major trauma patients in the experimental region were taken to the trauma centre, and by 1993 this had risen to only 39%. Crude death rates changed by the same amount in the control regions (46.5% in 1990-1 to 44.4% in 1992-3) as in the experimental region (44.8% to 41.3%). After standardisation, the estimated change in the probability of dying in the experimental region compared with the control regions was -0.8% per year (95% confidence interval -3.6% to 2.2%); for out of hours care, the change was 1.6% per year (-2.3% to 5.6%), and, for multiply injured patients, the change was -1.6% (-6.1% to 2.6%). CONCLUSION: Any reductions in mortality from regionalising major trauma care in shire areas of England would probably be modest compared with reports from the United States. PMID- 9402778 TI - Effect of a strict HLA matching policy on distribution of cadaveric kidney transplants to Indo-Asian and white European recipients: regional study. PMID- 9402779 TI - Improving uptake of breast screening in multiethnic populations: a randomised controlled trial using practice reception staff to contact non-attenders. AB - OBJECTIVES: To determine whether a two hour training programme for general practice reception staff could improve uptake in patients who had failed to attend for breast screening, and whether women from different ethnic groups benefited equally. DESIGN: Controlled trial, randomised by general practice. SETTING: Inner London borough of Newham. SUBJECTS: 2064 women aged 50-64 years who had failed to attend for breast screening. Women came from 26 of 37 eligible practices, 31% were white, 17% were Indian, 10% Pakistani, 14% black, 6% Bangladeshi, 1% Chinese, 4% were from other ethnic groups, and in 16% the ethnic group was not reported. MAIN OUTCOME MEASURES: Attendance for breast screening in relation to ethnic group in women who had not taken up their original invitation. RESULTS: Attendance in the intervention group was significantly better than in the control group (9% v 4%). The response was best in Indian women--it was 19% in the intervention group and 5% in the control group. CONCLUSIONS: This simple, low cost intervention improved breast screening rates modestly. Improvement was greatest in Indian women--probably because many practice staff shared their cultural and linguistic background. This intervention could be effective as part of a multifaceted strategy to improve uptake in areas with low rates. PMID- 9402780 TI - Work factors and upper limb disorders. PMID- 9402781 TI - Oxybutynin and cognitive dysfunction. PMID- 9402782 TI - ABC of palliative care. Depression, anxiety, and confusion. PMID- 9402783 TI - Disclosure of clinical audit records in law: risks and possible defences. PMID- 9402784 TI - New method for expressing survival in cancer. Use of percentage of "normal remaining life" may be confusing. PMID- 9402785 TI - New method for expressing survival in cancer. New method would be more meaningful to patients than 10 year survival rates. PMID- 9402786 TI - New method for expressing survival in cancer. Relation between survival times and patients' age at diagnosis must be taken into account. PMID- 9402787 TI - New method for expressing survival in cancer. Use of age specific relative survival is sufficient. PMID- 9402788 TI - Cognitive behaviour therapy. Review was unsystematic. PMID- 9402789 TI - Cognitive behaviour therapy. Patients are more likely to be treated with drugs. PMID- 9402790 TI - Patients with implants should be given implant cards for reference. PMID- 9402791 TI - General practice should be central to community mental health services. PMID- 9402792 TI - Treating diarrhoea. Proposals are irresponsible and impractical. PMID- 9402793 TI - Treating diarrhoea. Patients must be educated about which symptoms require treatment. PMID- 9402794 TI - Treating diarrhoea. Rehydration should have been emphasised. PMID- 9402795 TI - Treating diarrhoea. Diarrhoea is problem for animals too. PMID- 9402796 TI - Treating diarrhoea. Children deserve special attention. PMID- 9402797 TI - Treating diarrhoea. Advocating widespread use of antimicrobial agents has economic implications. PMID- 9402798 TI - Treating diarrhoea. More evidence is needed that widespread use of quinolones will benefit patients. PMID- 9402799 TI - Study linking enteroviral infection with motor neurone disease is not confirmed. PMID- 9402801 TI - Family secrets. Patients have right to privacy and confidentiality. PMID- 9402800 TI - Charity helps doctors with addictive diseases to obtain treatment. PMID- 9402802 TI - Family secrets. Most patients speak more freely when on their own. PMID- 9402803 TI - Nicotine replacement therapy on the NHS. Nicotine treatment cannot be regarded as replacement therapy. PMID- 9402804 TI - Nicotine replacement therapy on the NHS. Success rates of different smoking cessation treatments need to be compared. PMID- 9402805 TI - Old fashioned methods of diagnosis have their place. PMID- 9402806 TI - Reflex sympathetic dystrophy: fact and fiction. PMID- 9402807 TI - HIV management update. PMID- 9402808 TI - Mirtazapine revisited. PMID- 9402809 TI - Project Read--the importance of early learning. Rx: Read to your child. PMID- 9402810 TI - Preventing stroke in atrial fibrillation. AB - Atrial fibrillation, a common cardiac arrhythmia, is now recognized as a powerful risk factor for stroke. Previously, atrial fibrillation was thought to predispose persons to stroke only in the presence of rheumatic heart disease with mitral stenosis. The significant impact of nonvalvular atrial fibrillation on stroke incidence, recurrence and mortality was not fully appreciated. A series of clinical trials have confirmed that a five-fold increase in stroke incidence occurs in patients with atrial fibrillation, and that warfarin anticoagulation is efficacious in stroke prevention. This anticoagulation benefit was achieved with an acceptably low risk of serious hemorrhage. PMID- 9402811 TI - Paresthesias: a practical diagnostic approach. AB - Paresthesias may be caused by central or peripheral nervous system abnormalities. Central nervous system-induced paresthesias are most commonly caused by ischemia, structural or compressive phenomena, infection, inflammation or degenerative conditions. Peripherally induced paresthesias can be caused by entrapment syndromes, metabolic disturbances, trauma, inflammation, connective tissue diseases, toxins, hereditary conditions, malignancies, nutritional deficiencies and miscellaneous conditions. Confirming the diagnosis and establishing an etiology may require appropriate laboratory and radiologic studies, or other studies. In most cases, the specific clinical syndromes associated with the paresthesias, coupled with the presenting neurologic findings, provide the physician with a framework for the diagnosis. PMID- 9402812 TI - Complex regional pain syndrome. AB - The term "complex regional pain syndrome" encompasses causalgia and reflex sympathetic dystrophy. Symptoms of burning pain with autonomic and tissue changes begin shortly after an injury, usually to a distal extremity. The diagnosis is based on the history and the clinical findings. No confirmatory tests are available, although plain radiographs or a three-phase bone scan may be helpful in diagnosing some cases. Aggressive treatment, which may include sympathetic blockade, medications, physical therapy and psychotherapy, is essential for a favorable outcome. Despite treatment, many patients are left with varying degrees of chronic pain and disability. PMID- 9402813 TI - Drug treatment of migraine: Part II. Preventive therapy. AB - In most cases, successful preventive therapy for migraines requires daily medication for months or years. Perimenstrual use of a preventive agent is a common exception. Preventive therapy is usually undertaken in patients who have more than two headache episodes per month or those very much disabled by headaches. Beta blockers are usually the first choice for preventive therapy, and amitriptyline is also commonly used. Despite widespread use of calcium channel blockers for prevention of migraine, their benefits are controversial. Although effective for prevention of migraine, methysergide and phenelzine are usually relegated to last-resort use because of potentially serious side effects. The migraine patient who is refractory to standard preventive therapy may have rebound headache related to overuse of abortive migraine medications, or concomitant psychopathology. PMID- 9402814 TI - Occupational infections in health care workers: prevention and intervention. AB - Health care workers may be exposed to a variety of infections as they carry out their job responsibilities. Guidelines have been issued for prophylaxis following exposure to blood or body fluids known to be infected with the human immunodeficiency virus. Hepatitis B vaccine must be offered to all workers who may be exposed to blood and body fluids. Chemoprophylaxis is not available for workers exposed to hepatitis C. Health care facilities must conduct a tuberculosis risk assessment, provide skin testing at least yearly and develop isolation procedures for potentially infectious patients. The Occupational Safety and Health Administration currently mandates two-stage skin testing for all new employees at risk for tuberculosis exposure who have not had a skin test in the past year. Recent skin-test converters should be evaluated for isoniazid prophylaxis after a chest radiograph rules out active tuberculosis. Workers should be removed from the workplace from days 10 to 21 following exposure to varicella infection; vaccination of nonimmune workers should be considered. Because of possible side effects, the standard pertussis vaccine is not used in adults, but a new acellular pertussis vaccine has been effective in this group. PMID- 9402815 TI - AAP develops recommendations for the evaluation and treatment of HIV-exposed infants. PMID- 9402816 TI - First combination drug for patients with HIV. PMID- 9402817 TI - Prevalence of dry eye among the elderly. AB - PURPOSE: To study the demographics and estimate the prevalence of dry eye among elderly Americans. METHODS: A population-based prevalence study was performed in 2,520 residents of Salisbury, Maryland, aged 65 years and older as of September 1993. The population was derived from the Health Care Financing Administration Medicare database. After completing a standardized questionnaire pertaining to dry eye symptoms, 2,420 subjects underwent Schirmer and rose bengal tests and anatomic assessment of the meibomian glands. RESULTS: In this population, 14.6% (363/2,482) were symptomatic, defined as reporting one or more dry eye symptoms often or all the time; 2.2% (53/2,448) were symptomatic and had a low Schirmer test result (< or = 5 mm), and 2% (48/2,432) were symptomatic and had a high rose bengal test score (> or = 5). Furthermore, 3.5% (84/2,425) were symptomatic and had either a low Schirmer score or a high rose bengal score, and 0.7% (17/2,420) were symptomatic and had both a low Schirmer score and a high rose bengal score. No association of symptoms or signs was seen with age, sex, or race. Although anatomic features of meibomianitis were associated with the presence of symptoms (P = .01), 76% (67/88) of the individuals with these anatomic features were asymptomatic; 10.5% (260/2,480) reported that they currently use artificial tears or lubricants. CONCLUSIONS: Symptoms and signs of dry eye are common among the elderly but were not associated with age, race, or sex in this population-based sample of elderly Americans. Extrapolating to the United States population aged 65 to 84 years, the study yields an estimate of 4.3 million who experience symptoms of ocular irritation often or all the time. PMID- 9402818 TI - Use of high-speed, high-resolution thermography to evaluate the tear film layer. AB - PURPOSE: To evaluate the tear film layer in patients with dry eye and in normal subjects by measuring the corneal temperature with infrared radiation thermography. METHODS: One eye of each of 13 patients with dry eye and one eye of each of seven normal subjects were evaluated randomly. The corneal temperature was measured continuously with a recently improved infrared radiation thermography technique. We calculated the k value, which reflected the steepness of the corneal temperature change. The bigger the k value was, the more rapid was the decrease in corneal temperature, and this was directly related to increased evaporation. RESULTS: With normal blinking, the mean k value for patients with dry eye (5.6 +/- 2.9 per second) was significantly less than that in the control subjects (9.3 +/- 5.0 per second; P < .05). Keeping the eyes open after closing the eyes significantly decreased the k values compared with normal blinking in both groups (P < .05). CONCLUSIONS: Our findings demonstrate the usefulness of this method of measuring corneal temperature to evaluate the tear film layer. High-speed, high-resolution thermography detected subtle changes in corneal temperature with enhanced sensitivity and spatial and temporal resolution. We found that the mean k value, and therefore the rate of decline in corneal temperature in patients with dry eye, was significantly less than that in normal subjects. The k value may therefore reflect tear film layer stability. The measurement of the changes in the corneal temperature can thus give us valuable information on the tear film layer. PMID- 9402819 TI - Rose bengal staining and cytologic characteristics associated with lipid tear deficiency. AB - PURPOSE: To characterize ocular surface features of patients with an unstable tear film caused primarily by a lipid tear abnormality resulting from noninflamed meibomian gland dysfunction. METHODS: Retrospective clinical data and results from rose bengal staining, modified meibography, and impression cytology were reviewed in 78 patients (142 eyes), all of whom had normal tear secretion and clearance verified by the fluorescein clearance test but an unstable tear film evidenced by tear breakup time +/- SD of 3.4 +/- 2.1 seconds (normal, > 8 seconds). RESULTS: Of 201 symptoms, 147 (73%) were presumably caused by an unstable tear film, 46 (23%) resulted from inflammation, and none were diurnally worsened. All patients had meibomian gland dysfunction characterized by poor or no meibum expression, orifice squamous metaplasia, or acinar atrophy. Rose bengal staining was negative in 95 eyes (67%), positive on nonexposure zones in 30 eyes (21%), and positive on exposure zones in 17 eyes (12%). Among 90 eyes receiving impression cytology, six (7%) were normal, 49 (54%) had pure "lytic" changes characterized by disrupted cell-cell junctions of normal small cells in the nonexposure zone, three (3%) had pure squamous metaplasia without mucous aggregates, two (2%) had squamous metaplasia with mucous aggregates (the latter being a frequent finding of aqueous tear deficiency), and 31 (34%) were mixed with lytic changes and squamous metaplasia. CONCLUSION: Preferential distribution of rose bengal staining in the nonexposure zone and lytic cytologic changes without squamous metaplasia characterize lipid tear deficiency and help to differentiate it from aqueous tear deficiency in patients with an unstable tear film. PMID- 9402820 TI - Expression of a mucin-like glycoprotein produced by ocular surface epithelium in normal and keratinized cells. AB - PURPOSE: We previously characterized a monoclonal antibody (H185) to a mucin-like glycoprotein produced by human ocular surface epithelium. In the current study, we used H185 to investigate the pattern of the mucin-like glycoprotein in normal and keratinized apical cells of the ocular surface epithelium. METHODS: We compared the cell characteristics and the pattern of H185 binding in cytologically and immunohistochemically stained samples of apical cells of conjunctival surface epithelium from 20 normal subjects and six patients before and after treatment for superior limbic keratoconjunctivitis. RESULTS: In the superior bulbar conjunctiva of normal subjects, the intensity with which H185 antibody bound to apical surface epithelial cells varied, with areas of high-, medium-, and low-intensity binding occurring in a mosaic pattern. This mosaic pattern, and presumably expression of the mucin-like glycoprotein, was absent or remarkably reduced in keratinized cells obtained from patients with superior limbic keratoconjunctivitis before treatment. However, the pattern of H185 binding was normal in samples obtained 2 months after the start of treatment for superior limbic keratoconjunctivitis, and cells had recovered their normal small, round appearance. CONCLUSION: When they are keratinized, apical cells of the ocular surface epithelium are altered in appearance and lack the normal mosaic pattern of expression of a mucin-like glycoprotein. PMID- 9402821 TI - Gravity, blink rate, and lacrimal drainage capacity. AB - PURPOSE: To investigate the influence of gravity and blink rate on lacrimal drainage capacity and to learn whether lacrimal pump function can be measured with the drop test. METHODS: The drop test for lacrimal drainage capacity was performed in 20 test subjects, aged 12 to 30 years. Drops of a known volume of lukewarm saline solution were repeatedly instilled in the tear film for 3 minutes. Excessive saline solution was then removed, and the volume drained by the lacrimal passages was calculated. The drop test was performed both with the nasolacrimal duct in a 45-degree position and with the nasolacrimal duct in a horizontal position. The drop test was performed two times in each position, with the individual reading and not reading. A lower blink rate was induced by reading. RESULTS: There was a high correlation between blink rate and lacrimal drainage when the nasolacrimal duct was in a horizontal position. The volume drained with each blink was approximately 2 microliters. However, when gravity acted upon the fluid in the lacrimal sac-nasolacrimal duct in the direction of the tear flow, the lacrimal drainage capacity showed a significant but variable increase, and there was no significant correlation between blink rate and lacrimal drainage capacity. CONCLUSIONS: Lacrimal drainage capacity in young individuals was significantly affected by both blink rate and gravity. Lacrimal pump function can be measured quantitatively with the drop test. PMID- 9402823 TI - Changes in the anterior chamber configuration after small-incision cataract surgery with posterior chamber intraocular lens implantation. AB - PURPOSE: To report quantitative changes in the anterior chamber configuration after small-incision cataract surgery with implantation of a posterior chamber intraocular lens by means of ultrasound biomicroscopy. METHODS: We examined the anterior chamber configuration of 20 eyes of 20 patients before and 3 months after small-incision cataract surgery (phacoemulsification and aspiration plus foldable intraocular lens implantation through a 3.0- to 4.0-mm self-sealing wound) by means of ultrasound biomicroscopy. The following variables were measured: the anterior chamber depth at the center of the cornea, the angle opening distance 250 microns from the scleral spur (AOD250), the angle-opening distance 500 microns from the scleral spur (AOD500), and the trabecular-iris angle. RESULTS: The anterior chamber depth at the center of the cornea, AOD250, AOD500, and trabecular-iris angle increased significantly after surgery. The preoperative anterior chamber depth at the center of the cornea and trabecular iris angle were negatively correlated with the differences between the postoperative and preoperative values (P < .01). The preoperative values of all variables examined were negatively correlated with the ratios of the postoperative value to the preoperative value (P < .002). CONCLUSIONS: The present results showed that small-incision cataract surgery significantly deepened the anterior chamber and widened its angle. The more shallow the preoperative anterior chamber was, the greater the postoperative change of the chamber was; and the more narrow the preoperative angle was, the greater the postoperative change of the angle was. PMID- 9402822 TI - Amniotic membrane transplantation for conjunctival surface reconstruction. AB - PURPOSE: To determine whether preserved human amniotic membrane can be used to reconstruct the conjunctival defect created during surgical removal of a large lesion or during symblepharon lysis. METHODS: Amniotic membrane transplantation was performed in six consecutive patients (seven eyes) during removal of large conjunctival lesions and in nine patients (nine eyes) during removal of conjunctival scars or symblepharon. RESULTS: During a mean follow-up period +/- SD of 10.9 +/- 9.1 months (range, 2.2 to 34.0 months), 10 patients (11 eyes) showed successful surface reconstruction without recurrence, five patients (five eyes) showed improved visual acuity, and one patient (one eye) showed epithelialization within 3 weeks and resolution of motility restriction. Two patients (two eyes) showed partial success, with surrounding conjunctival inflammation. Three cases (three eyes) failed and exhibited recurrent scarring: one patient had received mitomycin treatment and beta radiation, whereas the transplanted amniotic membrane of the second patient was partially, and of the third patient was completely, dissolved or replaced by the inflamed pseudopterygial tissue. Two patients (two eyes) had epithelial cyst formation. CONCLUSION: Amniotic membrane transplantation can be considered an alternative substrate for conjunctival surface reconstruction during removal for large tumors, disfiguring scars, or symblepharon, especially for those whose surrounding conjunctival tissue remains relatively normal. PMID- 9402824 TI - Evaluation of Nd:YAG laser membranectomy in blocked tubes after glaucoma tube shunt surgery. AB - PURPOSE: To determine the effectiveness of Nd:YAG laser membranectomy for reopening blocked glaucoma tube shunts and maintaining the patency over time. METHODS: We reviewed retrospectively the records of 13 patients (13 eyes) who, during the period January 1990 through June 1996, underwent Nd:YAG laser membranectomy in an attempt to reopen a blocked glaucoma tube shunt. Intraocular pressure and tube patency were evaluated at each follow-up visit. RESULTS: Nd:YAG laser membranectomy effectively opened the blocked glaucoma tube shunts in 11 (84.6%) of 13 eyes. Two tubes could not be reopened. Reblockage occurred in seven eyes (53.8%) within the first 11 weeks; four tubes (30.8%) remained patent through follow-up periods of 39, 82, 106, and 169 weeks. Postlaser complications were moderate anterior chamber reaction in four eyes (30.8%), hyphema in two eyes (15.4%), corneal edema in two eyes (15.4%), pressure spike in one eye (7.7%), and shallow anterior chamber in one eye (7.7%). CONCLUSIONS: Nd:YAG laser membranectomy is effective in reopening blocked glaucoma tube shunts but is associated with a relatively high rate of subsequent reblockage in the initially successful cases. PMID- 9402825 TI - Endoscopic photocoagulation of the ciliary body for treatment of refractory glaucomas. AB - PURPOSE: To evaluate the safety and efficacy of endoscopic cyclophotocoagulation in the treatment of refractory glaucomas. METHODS: The preoperative and postoperative courses of 68 eyes of 68 patients who underwent endoscopic cyclophotocoagulation at our institution were retrospectively reviewed. Study patients had diverse forms of glaucoma, and most had failed maximal medical therapy as well as failed filtration or transscleral cyclodestructive procedures, or both. Endoscopic cyclophotocoagulation treatment encompassed 180 to 360 degrees of the ciliary body circumference and was performed through a limbal incision (56 eyes, 12 of which underwent concurrent cataract extraction) or pars plana incision (12 eyes). A second laser treatment was required in five eyes (7%). RESULTS: During the mean follow-up period of 12.9 months, mean +/- SD intraocular pressure decreased from 27.7 +/- 10.3 mm Hg preoperatively to 17.0 +/ 6.7 mm Hg at the final postoperative visit (P < .0001), for a mean reduction of 10.7 mm Hg and a mean percent decrease of 34%. Sixty-one eyes (90%) achieved an intraocular pressure < or = 21 mm Hg. Using this definition of success, Kaplan Meier analysis predicted a successful outcome in 94% of patients after 1 year and 82% after 2 years. The mean number of glaucoma medications used by each patient was reduced from 3.0 +/- 1.3 preoperatively to 2.0 +/- 1.3 postoperatively (P < .0001). Best-corrected visual acuity was stable or improved in 64 eyes (94%), with four (6%) losing 2 or more lines of Snellen acuity. No case of hypotony (intraocular pressure < 5 mm Hg) or phthisis was observed. CONCLUSION: These early results suggest that endoscopic cyclophotocoagulation is a safe and effective therapeutic modality for refractory glaucomas. PMID- 9402826 TI - Diode laser contact transscleral cyclophotocoagulation for refractory glaucoma in Asian patients. AB - PURPOSE: To evaluate the effectiveness and safety of diode laser contact transscleral cyclophotocoagulation in Asian patients with refractory glaucoma by lower energy settings with an innovative probe featuring a glass ball tip that focused the laser beam onto the ciliary body. METHODS: This prospective clinical study included consecutive Asian patients with dark irides and confirmed for glaucoma. Only one eye of each patient was treated. Diode laser contact transscleral cyclophotocoagulation treatment was performed with the center of the probe placed 1.5 mm behind the limbus. About 30 pulses of 810-mm laser radiation (power, 1.8 to 2.0 W; duration, 0.3 to 0.5 second) were applied around the eye. Patients were examined at fixed postoperative intervals. Intraocular pressure levels and postoperative complications were recorded. The relation between patient and disease characteristics, total laser energy delivered, and intraocular pressure effects were analyzed. RESULTS: Thirty-three patients were studied, with a mean follow-up period of 9.4 months. An average 56% of patients showed a 30% or greater drop in intraocular pressure. About 38% of patients achieved sustained intraocular pressure lowering to below 22 mm Hg at 18 months. Complications were few and included transient hypotony and iritis. CONCLUSIONS: In Asian patients with refractory glaucoma or painful glaucomatous eyes with poor visual acuity (defined for this study as worse than 20/200), low-energy-setting diode laser contact transscleral cyclophotocoagulation by means of the glass ball probe is relatively effective and safe. PMID- 9402827 TI - Quantitative differences between the optic nerve head and peripapillary retina in low-tension and high-tension primary open-angle glaucoma. AB - PURPOSE: To determine whether quantitative differences in optic nerve topography could be identified between patients having primary open-angle glaucoma with normal intraocular pressure (low-tension primary open-angle glaucoma [LT-POAG]) vs those with elevated intraocular pressure (high-tension primary open-angle glaucoma [HT-POAG]). METHODS: We attempted to match 31 eyes of 31 patients in the LT-POAG group on a case-by-case basis with comparable eyes of 31 patients with HT POAG. We used the Heidelberg Retina Tomograph to evaluate the optic nerve head and retinal nerve fiber layer. RESULTS: Cup areas and cup:disk area ratios were significantly larger (P < .05), whereas rim areas, rim volumes, retinal nerve fiber layer heights, and retinal nerve fiber layer cross-sectional areas were consistently smaller, but not significantly so (P > .05), in the LT-POAG group. The inferior neuroretinal rim area was significantly smaller (P < .05) and the mean deviation of superior arcuate area was significantly greater than the opposite sector in patients with LT-POAG but not in those with HT-POAG. A relationship between localized measurements of the optic nerve head and mean deviation was more apparent in the LT-POAG group than in the HT-POAG group. CONCLUSIONS: The optic cups were larger in patients with LT-POAG than in those with HT-POAG. Measurements of sectors of the optic disk correlated better with visual field changes in LT-POAG than did global measurements of the whole nerve head, indicating more vulnerability of the optic nerve to focal damage with low intraocular pressure. PMID- 9402828 TI - Langerhans cell histiocytosis with orbital involvement. AB - PURPOSE: To review three cases of Langerhans cell histiocytosis with orbital involvement that represent a significantly excessive incidence of this rare disease in one community. Current diagnostic criteria and therapeutic modalities related to Langerhans cell histiocytosis are reviewed. METHODS: Case reports. We present clinical, radiologic, histopathologic, and epidemiologic information on three patients with Langerhans cell histiocytosis. RESULTS: All three children, born within 18 months of one another, manifested rapidly progressive unilateral proptosis at age 2 years. By computed tomography, all had moderately enhancing lesions with involvement of the sphenoid bone and lateral orbit as well as the temporal lobe of the brain. All patients were treated with a combination of vincristine and prednisone, with variable resolution of their lesions. The occurrence of three cases in children born in Nogales, Arizona/ Mexico, suggests an incidence rate of 40 per million, which is approximately 26 times the expected rate (P = .0001). CONCLUSIONS: The extraordinarily high incidence and the concentration of cases in both time and space of this cluster implies that Langerhans cell histiocytosis may be a sentinel disease for unusual environmental exposures. PMID- 9402829 TI - Important concepts for treating ocular surface and tear disorders. AB - PURPOSE: To outline important concepts for treating ocular surface and tear disorders. METHOD: A review was conducted of recently published findings. RESULTS: Five concepts were delineated: ocular surface health is ensured by a close relationship between ocular surface epithelia and the preocular tear film; a stable tear film is inherently maintained by external adnexae; the intact protective mechanism is controlled by effective neuroanatomic integration; corneal epithelial stem cells are located at the corneoscleral limbus; and ocular surface epithelial cell function is supported by stromal fibroblasts and matrix. CONCLUSIONS: These concepts stress that ocular surface epithelia and the preocular tear film function as a unit and, furthermore, that several corneal and external diseases can be categorized as ocular surface and tear disorders. These concepts also help one formulate unified diagnostic and therapeutic strategies for management of these diseases. PMID- 9402830 TI - Treatment of fungal corneal ulcers with amphotericin B ointment. AB - PURPOSE: To report two patients with severe fungal corneal ulcers who were treated successfully with topical amphotericin B ointment. METHODS: Two eyes of two patients developed corneal ulcers and hypopyon after corneal foreign body removal. Aspergillus fumigatus and Fusarium solani were isolated in Patients 1 and 2, respectively. By antifungal susceptibility testing, amphotericin B was shown to have the lowest minimal inhibitory concentration. RESULT: Topical administration of amphotericin B ointment resulted in dramatic improvement in fungal corneal ulcers. CONCLUSIONS: Antifungal susceptibility tests may aid with selection of antifungal agents. Amphotericin B ointment is one of the promising therapies for keratomycosis caused by antimycotic-resistant fungi. PMID- 9402831 TI - Serum beta carotene, alpha tocopherol, and age-related maculopathy: the Blue Mountains Eye Study. AB - PURPOSE: To assess associations between serum beta carotene, alpha tocopherol, and age-related maculopathy. METHODS: We studied 156 subjects with age-related maculopathy matched for age, sex, and month of blood collection to 156 control subjects without age-related maculopathy. Subjects were identified from the Blue Mountains Eye Study: those with late age-related macular degeneration and early age-related maculopathy using examination and grading of retinal photographs, and control subjects without age-related maculopathy randomly sampled from the study population. RESULT: Neither serum alpha tocopherol nor beta carotene was significantly associated with age-related maculopathy. CONCLUSION: These findings provide no evidence of a protective association between serum alpha tocopherol or beta carotene and age-related maculopathy. PMID- 9402832 TI - Bilateral retinal vein occlusion associated with 5,10-methylenetetrahydrofolate reductase mutation. AB - PURPOSE: To report on the occurrence of 5,10-methylenetetrahydrofolate reductase (MTHFR) deficiency, known to cause mild to moderate hyperhomocystinemia and increased risk of vascular occlusive disease, in a young patient with bilateral central retinal vein occlusion. METHODS: A 25-year-old man was initially examined with central retinal vein occlusion in the right eye, followed 4 months later by a central retinal vein occlusion in the left eye. Studies for risk factors predisposing to thrombosis were performed. RESULTS: Hematologic studies failed to detect any pathology. However, the patient was found to be homozygous for 677C-T mutation in MTHFR enzyme. CONCLUSION: Central retinal vein occlusion may be associated with a mutation in MTHFR. PMID- 9402833 TI - Talc embolism: a static retinopathy. AB - PURPOSE: To document the static nature of crystalline deposits in talc retinopathy. METHOD: Case report. RESULTS: A 38-year-old woman with a remote history of intravenous drug abuse showed the incidental finding of fine, irregularly shaped refractile deposits in the retinal microvasculature. At 2-year follow-up, fundus findings were identical to the initial manifestation with respect to the distribution and position of crystalline deposits in each fundus. CONCLUSION: This case documents the static nature of crystalline deposits in talc retinopathy. PMID- 9402834 TI - Retinal toxicity associated with occupational exposure to the fish anesthetic MS 222. AB - PURPOSE: To report a case of retinopathy associated with chronic occupational exposure to ethyl-m-aminobenzoic acid methanesulfonate (MS-222), a retinotoxic fish anesthetic. METHOD: Case report with electroretinograms to document changes in visual electrophysiology. RESULTS: An ichthyologist with a long history of skin exposure to MS-222 was initially examined for decreased vision, photophobia, and photopsia. His electroretinogram abnormalities were similar to those seen in animal models of acute MS-222 toxicity. After terminating MS-222 contact for 7 months, his vision returned to normal, and his electroretinogram improved. CONCLUSION: Individuals with occupational exposure to MS-222 should exercise caution to avoid systemic absorption of this retinotoxic compound. PMID- 9402835 TI - Bilateral congenital optic nerve head pits in monozygotic siblings. AB - PURPOSE: To report bilateral congenital optic nerve head pits in monozygotic siblings. METHOD: Case reports. RESULTS: Pits were found in abnormally large optic disks in both eyes of two otherwise healthy female monozygotic siblings aged 15 years. Pit size increased and visual acuity decreased with increased optic disk area. In one eye, nonrhegmatogenous retinal detachment developed that eventually necessitated pars plana vitrectomy. The siblings' parents were unremarkable. CONCLUSIONS: Congenital optic nerve head pits can occur bilaterally in otherwise healthy monozygotic siblings with ophthalmologically unremarkable parents. Associated nonrhegmatogenous retinal detachment may be treated by pars plana vitrectomy. Pit size is positively correlated with disk area. PMID- 9402836 TI - Coats-type retinitis pigmentosa in a 4-year-old child. AB - PURPOSE: To describe the occurrence of Coats-like exudative retinopathy secondary to underlying retinitis pigmentosa in a 4-year-old child. METHOD: Case report. RESULTS: A 4-year-old girl had bilateral exudative retinal telangiectasia requiring photocoagulation. She subsequently developed progressive nyctalopia, photophobia, and reduced peripheral vision. Electroretinography and dark adaptometry at age 8 years confirmed the diagnosis of retinitis pigmentosa. CONCLUSIONS: Coats-like exudative retinopathy secondary to retinitis pigmentosa can manifest as early as age 4 years and can precede the diagnosis of the underlying retinal dystrophy. PMID- 9402837 TI - Trifocal uveal melanoma. AB - PURPOSE: To report the singular case of a patient who developed three noncontiguous uveal melanomas over a 30-year period. METHOD: Case report. RESULT: Systemic evaluation of a 50-year-old man with an iris melanoma and bilateral choroidal melanomas disclosed no evidence of metastases or other primary neoplastic disease. CONCLUSION: Although rare, the possibility of bilateral and multifocal uveal melanoma should be recognized. PMID- 9402838 TI - Bilateral subinternal limiting membrane hemorrhage with Terson syndrome. AB - PURPOSE: To report the anatomic location of bilateral dome-shaped posterior pole hemorrhages in a patient with Terson syndrome. METHODS: Case report. We performed bilateral vitrectomy for vitreous hemorrhage in a patient with Terson syndrome. After removal of vitreous hemorrhage, the tissue overlying a large discrete hemorrhage in the posterior pole was removed, and the tissue from one eye was examined histologically. RESULT: The discrete dome-shaped hemorrhage in the posterior pole was confined to the retina anteriorly by the internal limiting membrane. CONCLUSION: Large dome-shaped retinal hemorrhages with Terson syndrome can be located beneath the internal limiting membrane of the retina. PMID- 9402839 TI - Cavernous-dural fistula with secondary angle-closure glaucoma. AB - PURPOSE: To report a rare case of angle-closure glaucoma, secondary to the rapid development of a choroidal effusion, in a patient with a long-standing cavernous dural shunt. METHODS: Case report. Investigations included computed tomographic scan, magnetic resonance imaging, and carotid angiography. RESULTS: The development of the choroidal effusion occurred because of partial thrombosis of the ipsilateral superior ophthalmic vein and cavernous sinus. Drainage of the choroidal effusion resolved the angle-closure glaucoma. CONCLUSIONS: The combination of worsening signs and evidence of thrombosis indicates impending resolution of a cavernous-dural shunt. However, if a choroidal effusion causes angle-closure glaucoma, prompt surgical drainage should be considered to prevent permanent peripheral anterior synechiae formation, with the expectation that the effusion will not recur. PMID- 9402840 TI - Severe vision loss and neovascular glaucoma complicating superior ophthalmic vein approach to carotid-cavernous sinus fistula. AB - PURPOSE: To report a patient with unilateral vision loss and neovascular glaucoma after attempted superior ophthalmic vein embolization in the treatment of a carotid-cavernous sinus fistula. METHODS: A 69-year-old man with a history of a left dural carotid-cavernous sinus fistula underwent attempted treatment with superior ophthalmic vein embolization. The procedure was unsuccessful, and the left superior ophthalmic vein was ligated. RESULTS: Uncontrolled left proptosis and intraocular pressure necessitated urgent orbital decompression with severe vision loss and neovascular glaucoma. CONCLUSION: Superior ophthalmic vein embolization in the management of carotid-cavernous fistula may be associated with vision-threatening complications. PMID- 9402842 TI - Warthin tumor of the lacrimal gland. AB - PURPOSE: To describe a case of Warthin tumor involving the lacrimal gland. METHODS: Case report. The patient underwent a lateral orbitotomy to remove a nodular lesion involving the lacrimal gland. The specimen was submitted for histopathologic examination. RESULTS: The lesion was characterized by epithelial columnar cells arranged in solid nests or lining cystic spaces containing an exudative fluid and by pronounced lymphoid infiltrate with focal follicular formation. CONCLUSIONS: To our knowledge, this is the first case of Warthin tumor involving the lacrimal gland. Management requires complete excision of the lesion. PMID- 9402841 TI - Endometrial carcinoma in a patient with blepharophimosis syndrome and menstrual abnormality. AB - PURPOSE: To describe a woman with blepharophimosis syndrome and menstrual abnormality who developed endometrial carcinoma. METHODS: A 26-year-old woman with blepharophimosis syndrome and menstrual abnormality had increased serum luteinizing hormone and serum follicle-stimulating hormone levels. She was followed up for 5 years. RESULT: She developed endometrial carcinoma. CONCLUSIONS: Patients with blepharophimosis syndrome and menstrual abnormality should be examined gynecologically. Hormone-dependent endometrial carcinoma may develop. PMID- 9402843 TI - Non-African Burkitt lymphoma presenting with oral thrush and an orbital mass in a child. AB - PURPOSE: To report an unusual case of orbital involvement with non-African Burkitt lymphoma in a child. METHODS: A 26-month-old boy developed oral thrush and painless right proptosis with eyelid swelling. Magnetic resonance imaging disclosed a diffuse mass involving the inferior, lateral, and superior right orbit and both maxillary sinuses. Incisional biopsy of the orbital mass was performed. RESULTS: Histopathology disclosed diffuse infiltration of the orbital tissues by a high-grade lymphoid neoplasm that showed positive immunoreactivity for B-cell markers. Immunoglobulin gene rearrangement studies disclosed a characteristic t(8;14) translocation consistent with a small, noncleaved malignant lymphoma of the Burkitt type. CONCLUSION: Non-African Burkitt lymphoma may manifest with orbital involvement. PMID- 9402844 TI - Familial aggregation of age-related maculopathy. PMID- 9402846 TI - MTS1 expression and prognosis in acute myeloid leukemia. AB - MTS1, a tumor suppressor gene located on chromosome 9p21, has been shown to be altered in a number of human tumor cell lines, primary solid tumors, and leukemias. In this study we found low expression of MTS1 in lymphocytes from seven of nine healthy donors, but in none of eight granulocyte samples from the same controls, suggesting a physiological role of MTS1 in peripheral blood cells capable of proliferation, but not in end-stage differentiated cells. We detected MTS1 mRNA expression in 38 of 57 patients (66%) with acute myelogenous leukemia (AML) treated in a standardized clinical protocol. No deletion of the MTS1 gene was found in any of the AML samples tested. There was no significant association between expression of MTS1 and response to therapy, progression-free, or overall survival. Except for a negative correlation between MTS1 level and leukocyte count at diagnosis (p = 0.03), there was also no association with any of the known prognostic parameters in AML. We conclude that MTS1 shows a significantly higher expression in leukemic than in normal peripheral blood cells, that deletion of MTS1 is not a frequent event in AML, and that its expression is not significantly correlated with outcome of the disease. PMID- 9402845 TI - Randomized open label phase III trial of CEOP/IMVP-Dexa alternating chemotherapy and filgrastim versus CEOP/IMVP-Dexa alternating chemotherapy for aggressive non Hodgkin's lymphoma (NHL). A multicenter trial by the Austrian Working Group for Medical Tumor Therapy. AB - Primary end point of this trial was to reduce neutropenic infections during the treatment of aggressive NHL with CEOP/IMVP-Dexa (cyclophosphamide, epirubicin, vincristine, prednisolone ifosfamide, methotrexate, VP-16, and dexamethasone). Further, we studied the influence of filgrastim on dose intensity of CEOP/IMVP Dexa, on the rate of complete remissions, on the time to relapse, and on survival. Eighty-five patients with untreated large-cell NHL were randomized to one of two treatment arms; 74 patients were eligible. Thirty-eight patients in arm 1 were treated with CEOP/IMVP-Dexa chemotherapy and filgrastim, 36 in arm 2 with CEOP/IMVP-Dexa chemotherapy alone. In arm 1 filgrastim was self-injected by the patients at 5 micrograms/kg body wt. s.c. daily, except on the days when cytotoxic drugs were given. During treatment we did weekly complete blood counts. Median leukocyte counts were 10.91 x 10(9)/l and 5.46 x 10(9)/l in arm 1 and 2, respectively (p = 10(-6)). Median neutrophil counts were 7.7 x 10(9)/l in arm 1 and 2.72 x 10(9)/l in arm 2 (p < 10(-6)). Median neutrophil nadirs were 0.199 x 10(9)/l and 0.213 x 10(9)/l in arm 1 and 2, respectively (p = 0.09). Mean platelet nadirs were 95 and 152 x 10(9)/l (p = 0.000004) and mean hemoglobin nadirs 83.95 g/l and 92.78 g/l (p = 0.00558) in arm 1 and 2, respectively. Dose intensity of CEOP/IMVP-Dexa was 82.3% and 76.2% in arm 1 and 2, respectively (p = 0.041). Forty-two percent and 58% of patients experienced a febrile neutropenia in arm 1 and 2, respectively (not significant, NS). Median time to first neutropenic infection was in treatment week 11 and 6 in arm 1 and 2, respectively (NS). There was no significant difference in rate, duration, and kind of infection, duration of hospitalization, or antibiotic treatment. Seven toxic deaths occurred, all due to neutropenic infection, 6 and 1 in arm 1 and 2, respectively (p = 0.0732). Four of the six patients, who died of infection in arm 1 were older than 60 years. Complete remission rate was 83% and 66.7% in arm 1 and 2, respectively (NS). After a median observation time of 3 years there was no difference in time to relapse or survival. Filgrastim increases leukocyte and neutrophil counts and dose intensity, if used with CEOP/IMVP-Dexa chemotherapy in high-grade lymphomas. There was no significant effect on febrile neutropenia or infections. The more frequent fatal neutropenic infection rate in the filgrastim arm was not statistically significant. It is most appropriate to explain it by the patient's age in combination with the high dose intensity. The small increase in dose intensity had no effect on survival but probably decreased hemoglobin levels and platelet counts in arm 1. We were unable to show a benefit for filgrastim in combination with CEOP/IMVP-Dexa. PMID- 9402847 TI - In vitro down-regulation of bcl-2 expression by all-trans retinoic acid in AML blasts. AB - Using flow cytometry, we have investigated the effects of 0.5 microM all-trans retinoic acid (ATRA) on bcl-2 expression in the blast cells of 25 acute myeloblastic leukemia (AML) patients and the HL-60 cell line after incubation for 6 days. We observed a significant decrease of bcl-2 expression after treatment with ATRA in 12 of 25 AML samples and the HL-60 cells. The mean fluorescence intensity (MFI) ratio for the bcl-2 levels of the ATRA responders (n = 12) was reduced to 7.9 +/- 4.8 following incubation with ATRA compared with 10.9 +/- 6.5 (mean +/- SD) for control samples incubated without ATRA (p = 0.011). There was no significant difference between the baseline bcl-2 MFI ratio in the ATRA responders (11.14 +/- 7, n = 12) and the non responders (14.18 +/- 11.3, n = 13; p = 0.432). The down-regulation of bcl-2 expression by ATRA was not significantly associated with CD34-negative or -positive AML. There was no correlation between AML subtypes and regulation of bcl-2 expression by ATRA. Complete remission and overall survival were not significantly improved in bcl-2 down-regulated cases. Our data confirm that ATRA can down-regulate the bcl-2 expression in AML blasts. Because many chemotherapeutic agents also operate through the activation of programmed cell death and bcl-2 levels are positively associated with resistance to apoptosis, ATRA can be used in combination chemotherapy to increase the chemosensitivity of some patients with AML. PMID- 9402848 TI - Proliferative effect of postapheresis platelet donor plasma on a megakaryoblastic leukemia cell line. AB - Platelet donors sometimes show an increased platelet count following apheresis. We analyzed the effect of plasma obtained from such donors, along with thrombopoietin (Tpo), for growth and differentiation-inducing activity on a megakaryoblastic leukemia cell line, Meg-J. Colony formation was stimulated by donor plasma for Meg-J cells in a dose-dependent manner, and the time course of the sampling peaked 1 day after apheresis. Neither the donor plasma nor Tpo induced morphological differentiation. Donor plasma from any sampling time decreased CD41 and CD42b expression by more than 10%, and more than a 10% decrease of CD41 was induced by Tpo. The measurement of plasma Tpo showed no statistically significant increase after platelet donation. Our data suggest that, following apheresis, donor plasma contains factor(s) that: (a) stimulate growth rather than induce differentiation of a megakaryoblastic cell line; (b) are present in increased levels after apheresis; and (c) are different from Tpo. PMID- 9402850 TI - Prolonged fever as an unusual manifestation of the hyaline vascular type of Castleman's disease in the chest: report and review of the literature. PMID- 9402849 TI - Serum levels of interleukin-1 beta, luteinizing hormone, and prolactin correlate with the expression of CD45 isoforms on CD4+ peripheral blood T lymphocytes in healthy women. AB - The expected association between age and the CD45 isoforms expression on CD4+ T PBL is much more obvious in men than in women. We investigated whether or not circulating factors influence the differentiation of CD4+ T-PBL. Peripheral blood samples were obtained from 56 healthy age-matched subjects (28 men and 28 women, 21-55 years old). Mononuclear leukocytes were analyzed by three-color flow cytometry. The serum concentrations of interleukin-1 beta (IL-1 beta), interleukin-6, tumor necrosis factor-alpha (TNF-alpha), GM colony-stimulating factor, prolactin (Prl), and luteinizing hormone (LH) were determined by ELISA. The expected age-related decrease of naive (CD45RA+,RO-) cells and increase of memory (CD45RA-,RO+) cells among CD4+ T-PBL were observed in men only (p < 0.001 and 0.005). In women, these correlations were not significant. On the other hand, in women only, elevated IL-1 beta was associated with fewer naive and more memory cells among CD4+ T-PBL (p < 0.001). In both sexes, IL-1 beta correlated with the expression of CD25 on CD4+ T-PBL (on either naive or memory cells, p < 0.001). Other cytokines or the CD8+ T-PBL showed no significant correlation. In women, the elevation of LH at mid-cycle inversely correlated with the proportion of naive CD4+ T-PBL (p < 0.01). Elevated LH was associated with more CD25 on memory CD4+ T-PBL (p < 0.01). A significant correlation exists between IL-1 beta and LH (p < 0.001). Furthermore, in both sexes, Prl correlated with the proportion of CD4+ cells among T-PBL. In men, elevated Prl was associated with more naive CD4+ T-PBL (p < 0.005), while in women, Prl correlated with more transient CD45RA+, RO+ cells among CD4+ T-PBL and increased TNF-alpha (p < 0.05 for both). Thus, circulating IL-1 beta could be involved in the expression of CD25 on CD4+ T-PBL and favors the generation of memory CD4+ T-PBL. In women, the IL-1 beta- and/or mid-cycle-dependent processes seem to overwhelm the age-related changes. Elevated Prl might exert a dual influence: it favors the development of naive CD4+ T lymphocytes and possibly acts in, synergy with other cytokines during immune stimulation. PMID- 9402851 TI - Subcutaneous low-molecular-weight heparin for treatment of Trousseau's syndrome. AB - We report the case of a 76-year-old man with recurrent thromboses despite oral anticoagulation with phenprocoumon and low-grade chronic disseminated intravascular coagulation. Workup revealed a bronchial carcinoma (NSCCL) with hilar and mediastinal lymph node metastases. The clinical condition was consistent with Trousseau's syndrome. Based on reports in the literature, the therapy was changed to intravenous unfractionated heparin (UFH), which was effective in controlling the thrombotic coagulopathy. For practical reasons, despite a lack of evidence of its effectiveness in Trousseau's syndrome, therapy with UFH was changed to subcutaneous low-molecular-weight heparin (LMWH, nadroparine) in therapeutic doses of 100 IU/ kg body wt. 12 hourly. On an outpatient basis, five chemotherapy cycles were administered, and after metastases of the brain had been detected radiotherapy was initiated. Following 7.5 months of progressive neoplastic disease the patient died. He had remained free of thromboembolic complications under continued LMWH therapy during the last 6.5 months of his life. LMWH might be a convenient alternative to the established therapy with UFH in Trousseau's syndrome. PMID- 9402852 TI - Pancytopenia and fever of unknown origin in a 72-year-old woman. PMID- 9402853 TI - A 75-year-old woman with fatigue, monocytosis, and hypothyroidism. PMID- 9402854 TI - Mucosal tolerance and rheumatoid arthritis. PMID- 9402856 TI - Rheumatology--the whole is more than the sum of the parts... PMID- 9402855 TI - Musculoskeletal ultrasound imaging: a new diagnostic tool for the rheumatologist? PMID- 9402857 TI - Cryptic T-cell epitopes and their role in the pathogenesis of autoimmune diseases. AB - Immune recognition of self-proteins features prominently in the early pathogenesis of autoimmune rheumatic diseases such as rheumatoid arthritis (RA), Sjogren's syndrome (SS), systemic lupus erythematosus (SLE) and systemic sclerosis. The mechanisms which provide lymphocytes with access to such autoantigens are therefore fundamental in creating the opportunity for autoimmune responses to develop. It has long been thought that the tissue or cellular location of some self-proteins may determine that they are normally 'hidden' from immune recognition, thereby reducing their potential for autoantigenicity. Recently, this concept has been extended to apply even to different epitopes within the same protein. Many studies, encompassing a wide variety of antigens, have shown that some epitopes are not presented for recognition by T lymphocytes unless they are produced in unusually large concentrations or unless they are freed from the configuration of their native antigen. Epitopes for which this phenomenon occurs are described as cryptic. There is increasing interest in the possibility that crypticity may be an important characteristic of epitopes which are recognized by T lymphocytes in autoimmune pathogenesis. The evidence which has led to this theory and its significance are reviewed. PMID- 9402858 TI - Small fragments of cartilage oligomeric matrix protein in synovial fluid and serum as markers for cartilage degradation. AB - We determined the tissue distribution of cartilage oligomeric matrix protein (COMP) in man and evaluated COMP in synovial fluid (SF) and serum. COMP was purified from human articular cartilage. Polyclonal antibodies were used to detect COMP in tissue cryosections and protein extracts. COMP was determined quantitatively and qualitatively in SF and serum by competitive enzyme-linked immunosorbent assay and immunoblotting. Knee joint SF was taken from nine cadaveric and six living controls, 52 patients with osteoarthritis (OA), 85 patients with rheumatoid arthritis (RA) and 60 patients with other forms of inflammatory arthritis. The degradative potential of SF on native COMP was tested in vitro. The highest concentrations of COMP were measured in articular cartilage and meniscus, the lowest in rib and trachea. Compared with controls, the concentrations of COMP in SF and serum were elevated in 36 and 50% of the patients. A total of 84% of patients with RA and 60% of patients with other forms of inflammatory arthritis showed significant amounts of low-molecular-weight COMP fragments (50-70 kDa) in SF. In contrast, SF fragments were present in only 21% of the OA patients. Furthermore, 13% of SF taken from patients with RA or other forms of inflammatory arthritis were able to degrade COMP in vitro. Using inhibitors, the involvement of serine proteinases could be demonstrated in only 8% of the cases. Based on these results, the absolute levels of COMP in SF and serum, and its fragmentation pattern in SF, seem to be promising as markers of joint tissue metabolism. PMID- 9402859 TI - Tissue-derived macromolecules and markers of inflammation in serum in early rheumatoid arthritis: relationship to development of joint destruction in hands and feet. AB - We have previously shown that serum concentrations of cartilage oligomeric matrix protein (COMP) are increased early in rheumatoid arthritis (RA) patients who subsequently develop advanced large-joint destruction. A prognostic value for joint damage of serum concentrations of hyaluronan (HA) is also suggested by previous studies. In contrast, serum concentrations of bone sialoprotein (BSP) have not been useful for identifying patients with progressive large-joint destruction. In the present study, we have examined the hypothesis that serum concentrations of these tissue-derived markers are of prognostic value in RA for the development of radiographically detectable joint damage in hands and feet. Serum concentrations of COMP, HA and BSP were quantified in samples obtained from 62 patients within the first year after onset of RA and were related to the development of radiographically detectable damage in these joints after 5 yr. Neither the serum concentrations of COMP nor of BSP at inclusion predicted joint damage in hands and feet after 5 yr, and the concentration of these proteins did not change over the 5 yr period. However, the serum concentration of HA at inclusion correlated with the radiographic score at the 5 yr follow-up (r = 0.425, P < 0.01), but was not a better predictor in this respect than the erythrocyte sedimentation rate or C-reactive protein levels at inclusion. Thus, serum concentrations of the three studied tissue-derived macromolecules were in this study not useful for identifying patients prone to small-joint destruction. PMID- 9402860 TI - Different approaches to synovial membrane volume determination by magnetic resonance imaging: manual versus automated segmentation. AB - Automated fast (5-20 min) synovial membrane volume determination by MRI, based on pre-set post-gadolinium-DTPA enhancement thresholds, was evaluated as a substitute for a time-consuming (45-120 min), previously validated, manual segmentation method. Twenty-nine knees [rheumatoid arthritis (RA) 13, osteoarthritis (OA) 16] and 17 RA wrists were examined. At enhancement thresholds between 30 and 60%, the automated volumes (Syn(x%)) were highly significantly correlated to manual volumes (SynMan) (knees: rho = 0.78-0.91, P < 10(-5) to < 10(-9); wrists: rho = 0.87-0.95, P < 10(-4) to < 10(-6)). The absolute values of the automated estimates were extremely dependent on the threshold chosen. At the optimal threshold of 45%, the median numerical difference from SynMan was 7 ml (17%) in knees and 2 ml (25%) in wrists. At this threshold, the difference was not related to diagnosis, clinical inflammation or synovial membrane volume, e.g. no systematic errors were found. The inter-MRI variation, evaluated in three knees and three wrists, was higher than by manual segmentation, particularly due to sensitivity to malalignment artefacts. Examination of test objects proved the high accuracy of the general methodology for volume determinations (maximal error 6.3%). Preceded by the determination of reproducibility and the optimal threshold at the available MR unit, automated 'threshold' segmentation appears to be acceptable when changes rather than absolute values of synovial membrane volumes are most important, e.g. in clinical trials. PMID- 9402861 TI - Circulating soluble adhesion molecules in patients with classical polyarteritis nodosa. AB - The objective was to evaluate whether changes in circulating soluble adhesion molecule levels reflect disease activity in patients with systemic polyarteritis nodosa (PAN). A sandwich ELISA was used to measure soluble (s) intercellular adhesion molecule 1 (sICAM-1), vascular cell adhesion molecule 1 (sVCAM-1), E selectin, L-selectin and P-selectin in sera and plasma from 22 patients with active PAN, in sera from 13 of these patients taken serially during follow-up, and in sera from 13 healthy controls. At the time of diagnosis, sICAM-1, sVCAM-1 and sE-selectin levels (488.5 +/- 201.3, 1176.5 +/- 514.1 and 60.6 +/- 27 ng/ml, respectively) were significantly higher in patients than in controls (P < 0.0001, P = 0.001 and P = 0.003, respectively). In contrast, sL-selectin levels tended to be lower in patients than in controls. Within the first 7 days after starting treatment, there was a significant increase in sICAM-1 concentrations, which fell thereafter, but did not completely reach normal levels when patients achieved clinical remission. sE-selectin also remained elevated during follow-up. Only sVCAM-1 decreased, tending to reach normal values in inactive disease. Increased levels of sICAM-1, sVCAM-1 and sE-selectin, and decreased levels of sL-selectin, in active PAN suggest immune and endothelial stimulation during disease activity. Abnormal levels of soluble adhesion molecules in clinically inactive patients might reflect persistence of immune activation and/or endothelial cell exposure to a remaining inflammatory microenvironment. PMID- 9402862 TI - The association of HLA-DRB genes and the shared epitope with rheumatoid arthritis in Pakistan. AB - The association of particular HLA-DR alleles and the shared epitope with rheumatoid arthritis (RA) is now well established. The strength of these links varies between races. Furthermore, the proposition that the presence of the shared epitope is indicative of severe disease has been more difficult to sustain in non-Europeans. This study examines the frequency of HLA-DR and HLA-DRB1 amongst Pakistanis for the first time. Using the polymerase chain reaction (PCR) and sequence-specific oligonucleotide probes (PCR-SSOP) and primers (PCR-SSP), HLA-DR phenotype and genotype frequencies were ascertained in 86 RA hospital out patients and 79 healthy controls matched for age, gender and ethnicity. HLA-DR1 and HLA-DR4 frequency was similar in patients and controls. HLA-DR10 occurred in 26 instances (15%) in RA and in eight (5%) controls (Pcorr = 0.048). HLA-DR2 was also increased in patients (P = 0.053) and its major subtype DR15 was significantly increased (Pcorr = 0.03). HLA-DR5 frequency was 5% in patients and 19% in controls (Pcorr = 0.002). The HLA-DR4 alleles possessing the shared epitope were more common in RA (Pcorr = 0.03) and this difference was enhanced by inclusion of other alleles possessing the shared epitope (Pcorr = 0.002). Shared epitope alleles were observed in 43 (50%) patients and 17 (22%) controls (Pcorr = 0.003). The shared epitope did not distinguish patients with more severe disease, as reflected by pain, joint deformities, disability, rheumatoid factor or X-ray damage. The distribution of HLA-DR alleles in Pakistanis with RA supports the shared epitope hypothesis. In common with other non-European racial groups, HLA DR4 was not associated with RA. Unlike other groups, there was a weak link of RA with HLA-DR2. A protective effect of HLA-DR5 was apparent. In accord with some other studies, the shared epitope in this hospital out-patient population was not a marker for more severe disease. PMID- 9402863 TI - Mesangial glomerulonephritis as an extra-articular manifestation of rheumatoid arthritis. AB - Mesangial glomerulonephritis (MesGN) is a frequent renal biopsy finding in patients with rheumatoid arthritis (RA) presenting with haematuria and/or proteinuria. The purpose of this study was to describe the mesangial immunofluorescence (IF) findings in 37 such RA patients, and to correlate the nature of the glomerular immunodeposits with clinical data, levels of serum immunoglobulins and the concentrations of serum rheumatoid factors (RFs). The serological findings in RA patients with MesGN were correlated with those of RA patients matched for age, sex and duration of RA, but without clinically evident renal disease. The most characteristic mesangial IF finding in 37 RA patients with MesGN was IgM observed in 29 specimens (78%). IgA (n = 16) and C3 (n = 15) deposits were also frequently found, whereas C1q (n = 3) and IgG (n = 2) deposits were only occasionally seen. Two main patterns of IF findings could be classified. (1) Granular IgM deposits as a sole or main finding (IgM GN; 25 specimens). The intensity of the mesangial IgM deposits did not correlate with the duration or severity of RA nor with the levels of serum immunoglobulins. However, a significant correlation with the intensity of mesangial IgM deposits and the levels of serum IgM class RF was observed. (2) Granular IgA deposits usually together with the complement component C3, i.e. IgA glomerulonephritis (IgA GN; nine specimens). The intensity of mesangial IgA deposits correlated significantly with the duration and severity of RA, and especially with serum IgA levels. Both RA patients with IgA GN or IgM GN were associated with a more frequent occurrence and higher titres of serum RFs when compared with the RA patients without nephropathy. The clinicopathological correlations and the association with RF support the concept that MesGN is related to the basic rheumatoid disease, and it should be regarded as an extra-articular manifestation of RA. PMID- 9402864 TI - The rational use of methotrexate in rheumatoid arthritis and other rheumatic diseases. AB - Methotrexate's mechanism of action affects both the inflammatory and immunosuppressive aspects of response. Its kinetics are defined and include variable absorption, intracellular metabolism, and both renal and biliary excretion. Methotrexate is clearly effective in the treatment of rheumatoid arthritis and may be able to decrease the rate of formation of new bony erosions. It is also effective in psoriatic arthritis and is being used in a multiplicity of other rheumatic diseases. The most common toxicities ascribed to methotrexate are gastrointestinal (e.g. stomatitis) and central nervous system (e.g. headache, fatigue, malaise). Methotrexate-induced hepatic cirrhosis is less common in rheumatoid arthritis than previously thought, although its occurrence in psoriasis is probably higher than in rheumatoid arthritis. Haematological, renal and pulmonary toxicity occur, but are rare, while teratogenicity is well documented. A new and disturbing adverse event, pseudolymphomas are being reported at present. PMID- 9402865 TI - Rheumatology in Switzerland. PMID- 9402866 TI - Mycobacterium marinum infection causing septic arthritis and osteomyelitis. AB - A 48-yr-old female on immunosuppressive therapy for fibrosing alveolitis and polymyositis developed a septic arthritis of the left middle finger proximal interphalangeal joint, tenosynovitis of the left palm and osteomyelitis of the right hindfoot due to infection with Mycobacterium marinum. Such widespread and severe bone and joint involvement has not been described previously with this organism. PMID- 9402867 TI - Polyarteritis nodosa and the antiphospholipid syndrome. AB - We describe a case of classical polyarteritis nodosa (PAN) with visceral aneurysms presenting with renal infarction and hypertension. The female patient also had all the laboratory features of the antiphospholipid syndrome (APS) and 2 months into her illness developed a large iliofemoral thrombosis. She responded well to immunosuppressive therapy and anti-coagulation. Repeat arteriogram showed regression of the visceral aneurysms. The link between PAN and APS, and the therapeutic dilemma posed by this association, are discussed. PMID- 9402869 TI - School attendance and juvenile chronic arthritis. AB - We studied the school attendance of 113 children and adolescents (mean age 11 yr, S.D. 3.8, range 3-18 yr) with juvenile chronic arthritis (73 with pauci- and 40 with polyarthritis). The mean attendance rate for the group was 92% (equivalent to 15 absent days a year) with a median of 97%. Attendance was significantly lower in the more severely affected poly group (90% vs 98% in the pauci group; P = 0.03). We found associations of school absence (i) with decreased compliance with physical treatments (r = -0.35, P < 0.05 for compliance with physiotherapy) in the poly group and (ii) with child psychological deviance (r = 0.36 for parentally rated and r = 0.42 for teacher-rated psychological deviance; both P < 0.05) in the pauci group. We conclude that school attendance can be good in severely affected children. Severity of illness, treatment compliance and psychological problems in the child may affect school attendance. PMID- 9402868 TI - Juvenile chronic arthritis: diagnosis and management of tibio-talar and sub-talar disease. AB - The aim of this study was to compare clinical evaluation of the site of hindfoot synovitis with contrast-enhanced magnetic resonance imaging (MRI) findings in children with juvenile chronic arthritis (JCA), and to evaluate the efficacy of selective guided intra-articular steroid injections. Thirteen symptomatic ankles of 11 consecutive JCA patients were examined clinically and with contrast enhanced MRI. Pannus was demonstrated on MRI in both tibio-talar and sub-talar joints in 10 ankles, in the tibio-talar joint only in one ankle and in neither joint in two ankles. Correlation of clinical and MRI findings was good for the tibio-talar joint with concordance in 11/13 cases. Correlation was poor for the sub-talar joints. Of the 10 sub-talar joints shown to have pannus on MRI, only two were thought to have had definite clinical evidence of synovitis. Guided intra-articular steroid injection resulted in at least 6 months remission in 6/9 ankles compared with 1/10 ankles which had had previous unguided injections. We therefore recommend the use of image guidance for intra-articular triamcinolone hexacetonide injection in children with hindfoot synovitis. PMID- 9402870 TI - Pachydermoperiostosis in childhood. AB - We report a family with pachydermoperiostosis (idiopathic hypertrophic osteoarthropathy) spanning four generations with 10 affected individuals, four of whom are children although pachydermoperiostosis is rare in childhood. In this family, with intermarriage, the inheritance is autosomal recessive and it is possible that there are individuals who are homozygous for the pachydermoperiostosis gene. These individuals do not appear to be more severely affected, although one of them had a cleft palate and congenital heart defect which may be a manifestation of being homozygous. PMID- 9402871 TI - Protracted arthritis of familial Mediterranean fever (an unusual complication). AB - An unusual case of familial Mediterranean fever and vasculitis in which the patient developed amyloidosis and had protracted arthritis persisting for years is presented. The long-standing arthritis did not respond to corticosteroid and colchicine therapy, but an excellent response to synovectomy was achieved. PMID- 9402872 TI - Clinical improvement and radiological deterioration in rheumatoid arthritis. PMID- 9402873 TI - The use of granulocyte colony-stimulating factor in joint replacement surgery in a patient with Felty's syndrome. PMID- 9402874 TI - Gastrointestinal complications and meloxicam. PMID- 9402875 TI - Pre-treatment X-rays for rheumatoid arthritis treated with methotrexate. PMID- 9402876 TI - Anti-beta 2-glycoprotein I antibody testing in patients with antiphospholipid syndrome. PMID- 9402877 TI - Tibiofemoral joint space narrowing and meniscal lesions. PMID- 9402878 TI - Kikuchi's lymphadenitis developing in a patient with mixed connective tissue disease and Hashimoto's thyroiditis. PMID- 9402879 TI - Iliopsoas bursa enlargement. PMID- 9402880 TI - Mannose-binding lectin gene polymorphism in Greek systemic lupus erythematosus patients. PMID- 9402881 TI - EEG coherency. I: Statistics, reference electrode, volume conduction, Laplacians, cortical imaging, and interpretation at multiple scales. AB - Several methodological issues which impact experimental design and physiological interpretations in EEG coherence studies are considered, including reference electrode and volume conduction contributions to erroneous coherence estimates. A new measure, 'reduced coherency', is introduced as the difference between measured coherency and the coherency expected from uncorrelated neocortical sources, based on simulations and analytic-statistical studies with a volume conductor model. The concept of reduced coherency is shown to be in semi quantitative agreement with experimental EEG data. The impact of volume conduction on statistical confidence intervals for coherence estimates is discussed. Conventional reference, average reference, bipolar, Laplacian, and cortical image coherencies are shown to be partly independent measures of neocortical dynamic function at different spatial scales, due to each method's unique spatial filtering of intracranial source activity. PMID- 9402882 TI - Correlation of developmental neurological findings with spectral analytical EEG evaluations in pre-school age children. AB - For the differentiation of developmental neurological disorders in pre-school age children, the relationship between automatically derived EEG parameters and developmental neurological findings was investigated. Within the scope of the Munich Pediatric Longitudinal Study, the sample sets of 4- and 5-year-old children (according to the frontal and parieto-occipital EEG derivations) with selected abnormal findings categorized by special items were compared with the corresponding control groups. This was carried out by means of one-sided t tests and relative frequency band-related as well as single-step spectral power parameters in the alpha range of the EEG. Automatic analysis using single-step power values was superior to that using band-related parameters. This led to the conclusion that use of age-specific single-step parameters for a quantitative EEG analysis and ignoring the classical frequency bands will yield statistically greatly improved results. For 4- and 5-year-old children, the best separation of the neurologically abnormal groups from the normal control groups was obtained using relative spectral values in the frequency range of 9.0-9.8 Hz with a maximum at 9.4 Hz. At the same time, the topographical conditions of brain immaturation should be taken into account. The results for the children examined in this study differ in a stronger distinction over the frontocentral brain region of 4- and 5-year-olds (P < 0.01) and through an additional distinction over the parietooccipital region of the 5-year-olds (P < 0.001). It still must be tested whether the spectral parameter at 9.4 Hz is age-specific for 4- and 5-year old children or whether in other age groups different spectral parameters are of use. As an examiner-independent method, the automatic EEG analysis should become an integral component of developmental neurological diagnostics. PMID- 9402883 TI - Quantitative analysis of discontinuous EEG in premature and full-term newborns during quiet sleep. AB - To assess the spatio-temporal structure of discontinuous EEG tracing in mature and immature newborns, we analysed mean spectral power in frequency bands between 0.8 and 16.8 Hz in 6 full-term newborns and 7 premature newborns < 32 weeks of conceptional age. The most striking results showed a significantly higher mean spectral power for the first half of bursts than for the second half recorded in > 2.8-14.8 Hz frequency bands. This pattern was more pronounced in premature than in full-term newborns. No clear differences were observed in comparisons between the first and the second half of the interburst periods. In addition, as far as mid and high frequency band spectra were considered, the mean spectral power of burst was, in both groups, higher in the right as compared to the left occipital regions. PMID- 9402885 TI - FFT analysis of EEG during stage 2-to-REM transitions in narcoleptic patients and normal sleepers. AB - The EEG in REM sleep is markedly different from the EEG in non-REM sleep; however, very little research has been conducted analyzing the transition from NREM to REM sleep. The purpose of this study was to describe the changes in EEG power spectra that coincide with significant physiological events throughout the transition from stage 2 to REM sleep. Furthermore, a comparison of the stage 2 to REM transition between narcoleptic patients and normal sleepers was conducted. Five female and five male patients diagnosed with narcolepsy-cataplexy and 10 normal sleepers matched for age and gender participated. Analyses were based on the consecutive appearance of 3 common physiological indicators of REM sleep: First, a decrease in tonic submental EMG activity; second, the appearance of saw tooth waves; and third, the appearance of rapid eye movements. Systematic changes in EEG power density were evident for delta, theta, alpha and sigma frequencies, across the stage 2-REM transition. These changes appeared to be relatively continuous throughout this transition rather than changing dramatically following the onset of any one of the 3 primary physiological indicators of REM sleep. Furthermore, the transition from stage 2 to stage REM appeared to be similar for narcoleptic patients and normal sleepers. PMID- 9402884 TI - Spindle frequency activity in the sleep EEG: individual differences and topographic distribution. AB - The brain topography of EEG power spectra in the frequency range of sleep spindles was investigated in 34 sleep recordings from 20 healthy young men. Referential (F3-A2, C3-A2, P3-A2 and O1-A2) and bipolar derivations (F3-C3, C3-P3 and P3-O1) along the anteroposterior axis were used. Sleep spindles gave rise to a distinct peak in the EEG power spectrum. The distribution of the peak frequencies pooled over subjects and derivations showed a bimodal pattern with modes at 11.5 and 13.0 Hz, and a trough at 12.25 Hz. The large inter-subject variation in peak frequency (range: 1.25 Hz) contrasted with the small intra subject variation between derivations, non-REM sleep episodes and different nights. In some individuals and/or some derivations, only a single spindle peak was present. The topographic distributions from referential and bipolar recordings showed differences. The power showed a declining trend over consecutive non-REM sleep episodes in the low range of spindle frequency activity and a rising trend in the high range. The functional and topographic heterogeneity of sleep spindles in conjunction with the intra-subject stability of their frequency are important characteristics for the analysis of sleep regulation on the basis of the EEG. PMID- 9402886 TI - A normative study of the maintenance of wakefulness test (MWT). AB - The maintenance of wakefulness test (MWT) is a daytime polysomnographic procedure which quantifies wake tendency by measuring the ability to remain awake during soporific circumstances. We present normative data based on 64 healthy subjects (27 males and 37 females) who adhered to uniform MWT procedural conditions including polysomnographic montage, illuminance level, seating position, room temperature, meal timing, and subject instructions. When allowed a maximum trial duration of 40 min, subjects' mean sleep latency to the first epoch of sustained sleep was 35.2 +/- 7.9 min. The lower normal limit, defined as two standard deviations below the mean, was 19.4 min. Calculation of data on the basis of a maximum trial duration of 20 min and sleep latency to the first appearance of brief sleep (a microsleep episode or one epoch of any stage of sleep) yielded a mean sleep latency of 18.1 +/- 3.6 min and a lower normal limit of 10.9 min. Sleep latency scores were significantly higher than those previously reported in patients with disorders of excessive somnolence. Therefore, the MWT appears to be a useful procedure in differentiating groups with normal daytime wake tendency from those with impaired wake tendency and in identifying individuals with pathologic inability to remain awake under soporific circumstances. PMID- 9402887 TI - G-force induced alterations in rat EEG activity: a quantitative analysis. AB - A major physical limitation affecting pilots is G-force (+Gz, head-to-foot inertial load) induced loss of consciousness. Previous studies have shown that +Gz produces qualitatively similar effects on human and rat EEG activity. The present study sought to quantitatively correlate changes in rat EEG activity with increasing +Gz levels. A frontal-parietal differential electrode recorded rat EEG data during +Gz exposures (30 s) ranging from +0.5 to +25.0 Gz. Acceleration levels < or = +10 Gz had little effect on EEG activity. Acceleration levels of +15 to +20 Gz were associated with increased EEG slowing, depression and sharp waves. Acceleration levels > or = +17.5 Gz evoked burst suppression followed by isoelectric activity. Times to first onset of delta, depressed, and isoelectric EEG activity were approximately 12, 14 and 18 s, respectively. Acceleration effects on delta (1-4 Hz), theta (5-8 Hz), alpha (9-12 Hz), beta (13-30 Hz) and total (1-30 Hz) EEG powers were examined using Fourier transform analysis. EEG measures with the most predictive value at the following post-acceleration onset times (PAOT) were as follows (in s); increasing theta power: PAOT 0-2, decreasing delta power: PAOT 3-9, and decreasing beta power: PAOT > or = 12. This study provides a quantitative description of +Gz-induced alterations in EEG magnitude, time course and spectral content. Additionally, several EEG measures were identified which correlated with acceleration level at specific post-acceleration onset times. PMID- 9402888 TI - Somatosensory evoked potential monitoring in carotid surgery. I. Relationships between qualitative SEP alterations and intraoperative events. AB - This paper presents the results of intraoperative median nerve SEP monitoring in 205 successive patients undergoing isolated carotid endarterectomy (CE) (N = 172) or CE followed by coronary bypass (CBP) and/or vascular replacement (VR) (N = 33). The left and right median nerves were alternately stimulated and recordings performed on 4 channels: cervical, ipsi- and contralateral parietal, and frontal. SEPs were qualitatively rated in terms of mild, moderate, or severe ipsilateral, contralateral, or bilateral abnormalities. The SEP abnormalities were subdivided into 5 categories as a function of their relationships with intraoperative events: no alterations (67.3%), early or late SEP alterations after carotid cross clamping (15.6%), SEP alterations after a drop in blood pressure (occurring outside of or within the cross-clamping period) (15.1%), SEP alterations of a most likely embolic origin (2.4%), SEP changes after head positioning (1%), and SEP changes after a modification of the anesthetic regimen (1.5%). Only moderate to severe SEP alterations occurring soon after carotid cross-clamping justified shunt installation in 16% of the cases. SEP alterations after a drop in blood pressure were reversed merely by restoring blood pressure. The neurological outcome was uneventful in 94.2% of cases. Of the 12 patients who developed neurological sequellae, only one case presented transient sequellae after isolated CE without SEP changes while most cases either had undergone combined CE and CBP and/or VR (6 cases) or had presented SEP alterations of embolic origin (3 cases). We conclude that our system of qualitative rating of SEPs proved very sensitive to intraoperative hemodynamic disturbances or macroembolisms. PMID- 9402889 TI - Long sensory tracts (cuneate fascicle) in cervical somatosensory evoked potential after median nerve stimulation. AB - Low amplitude high frequency waves (LHW) were investigated in normal and patient cervical somatosensory evoked potentials after median nerve stimulation (CSEP) in parallel to normal and patient conducted somatosensory evoked potentials (SEP) after tibial nerve stimulation. Normal recordings were obtained in five subjects undergoing dorsal root entry zone (DREZ) coagulation for pain relief. Patient recordings were obtained in 11 subjects suffering from either syringomyelia, spinal cord tumour, or both. All recordings were made intraoperatively from the dorsal spinal cord surface using the subpial recording technique. Normal CSEP showed typical triphasic potential starting with an initial P9, followed by N13 and a final positivity, P1. Numerous LHW were superimposed on slow triphasic potential. To improve the visibility of LHW, slow triphasic potential was removed from the original CSEP. Potentials thus obtained contained only high frequency components of CSEP, i.e. LHW. They were compared with conducted SEP after tibial nerve stimulation. Comparison revealed similarities in high frequency, low amplitude and general wave form, LHW thus showing characteristics of conducted potential. Duration was found to be significantly shorter than normal duration in both patient LHW (Student's t-test, P < 0.0005) and patient conducted SEP (Student's t-test, P = 0.064). A shorter duration was associated with worsening of configuration in patient LHW and patient conducted SEP. These changes of LHW could not be connected with distortion of N13 seen in patient CSEP. A shorter duration and worsening of configuration in patient LHW were most prominent in cases with a loss of vibration and posture senses, but were also observed in cases where only pain and temperature senses were affected. We therefore concluded that cuneate fascicle is the most likely generator of LHW, although the participation of other cervical long sensory tracts, e.g. spinothalamic tract, cannot be ruled out. PMID- 9402890 TI - Somatosensory evoked magnetic fields to median nerve stimulation: interhemispheric differences in a normal population. AB - The objective of the present study was to evaluate the normal interhemispheric variability of the locations and activation strengths of the somatosensory cortices. Somatosensory evoked magnetic fields (SEFs) were recorded with a 122 channel magnetometer in 23 healthy subjects (mean age 57 years) to stimulation of left and right median nerves. Equivalent current dipole (ECD) strengths and locations were determined for the main SEF deflections at the contralateral primary sensorimotor (SMI) and secondary somatosensory (SIIc) cortices. In a Cartesian co-ordinate system, defined by the preauricular points and the nasion, the SMI sources were slightly but significantly more laterally and anteriorly located in the right than in the left hemisphere. No systematic co-ordinate asymmetries were found for the SIIc sources. In individual subjects, the interhemispheric differences in the ECD co-ordinates averaged less than 6 mm at both SMI and SIIc. The group means of the source strengths did not differ between the hemispheres, but individual differences were on average 20% for the SMI and 65% for the SIIc sources. We conclude that at the individual level, the median nerve SEFs from SMI can be used to detect abnormally large interhemispheric asymmetries of source locations in the centimetre scale. PMID- 9402891 TI - P300 from a single-stimulus paradigm: passive versus active tasks and stimulus modality. AB - The P300 component of the event-related brain potential (ERP) was elicited with auditory and visual stimuli in separate experiments. Each study compared an oddball paradigm that presented both target and standard stimuli with a single stimulus paradigm that presented a target but no standard stimuli. Subjects were instructed in different conditions either to ignore the stimuli, press a response key to the target, or maintain a mental count of the targets. For the passive ignore conditions, P300 amplitude from the single-stimulus paradigm was larger than that from the oddball paradigm. For the active tasks, P300 amplitude from the oddball paradigm was larger than that from the single-stimulus paradigm. For the press and count conditions, P300 amplitude and latency were highly similar for the oddball and single-stimulus procedures. The findings suggest that the single-stimulus paradigm can provide reliable cognitive measures in clinical/applied testing for both passive and active response conditions. PMID- 9402892 TI - An overview of age-related changes in the scalp distribution of P3b. AB - In this overview of 7 studies, the scalp distribution of the P3b component (i.e. the P3 or P300) of the event-related potential elicited by target events in young and older adults was assessed. The target P3b data were recorded in either auditory oddball paradigms or in visual study tasks in which orienting activity was manipulated (as a within-subjects variable) in investigations of indirect memory. Some of the studies required choice reaction time responses, whereas others required responses only to the target stimuli. Motor response requirements had a profound effect on the P3b scalp distribution of older but not of younger subjects. The presence of a frontally oriented scalp focus in the topographies of the older adults in most of the tasks described here is consistent with older adults continuing to use prefrontal processes for stimuli that should have already been well encoded and/or categorized. However, although older subjects generally had different P3b scalp distributions than younger subjects, their scalp distributions were modulated similarly by task requirements. These data suggest that similar mechanisms modulate the scalp distribution of P3b in older compared to younger adults. However, in the older adult, these scalp distribution changes in response to task demands are superimposed on a frontally oriented scalp focus due to a putative frontal lobe contribution to target P3b topography. PMID- 9402893 TI - Differential changes of laser evoked potentials, late auditory evoked potentials and P300 under morphine in chronic pain patients. AB - The present study investigates the differential behavior of laser evoked brain potentials (LEPs), late auditory evoked potentials (AEP) and the endogenous P300 in response to morphine treatment, examined in 6 chronic pain patients. The main result was that in parallel with marked clinical pain relief, amplitudes of the long latency LEP positivity (P400) were significantly reduced under morphine. One patient suffering from extremely painful osteoporosis for 20 years exhibited a large middle latency component (N170) which was prominently attenuated by morphine. In contrast to LEP amplitude reductions, auditory N1 and P2 potentials appeared either unchanged or even enlarged during morphine treatment. Also P300 amplitude was slightly increased under morphine. Reaction time and mood scales also failed to indicate any sedation. Obviously, LEPs reflected specifically the analgesic morphine effect in this study, while stability or enhancement of AEPs and P300 during morphine treatment indicated lack of sedation or even improved perception and concentration due to the removal of persistent pain as a disruptive perceptual-cognitive stressor. PMID- 9402894 TI - Short latency vestibular evoked potentials (VsEPs) to linear acceleration impulses in rats. AB - In this study, short latency (t < 12.7 ms) vestibular evoked potentials (VsEPs) in response to linear acceleration impulses were recorded in 37 rats. A new technique (based on a solenoid) was used for generating linear force impulses that were delivered to the animal's head. The impulse had a maximal peak acceleration of 12 g. During the impulse, the displacement was 50 microns (at 4 g) and the rise time was 1.0 ms. A stimulation rate of 2/s was usually used. The VsEPs (averaged responses to 128 stimulations, digital filter: 300-1500 Hz) were recorded with electrodes on pinna and vertex, and were composed of 4-6 clear waves with mean amplitudes (for a 4 g stimulus) of 1-5 microV. The VsEPs were resistant to white noise masking, and were significantly suppressed (P < 0.05) following bilateral application of a saturated KCl solution to the inner ear, showing that contributions of the auditory and somatosensory systems are negligible. The latency of the response decreased as a power law function of stimulus magnitude, and the amplitude of the first wave increased as a sigmoid function of stimulus magnitude. VsEP responses were still present at the lowest intensities attainable (0.06-0.4 g) and reached saturation at 9 g. The amplitude of the later components was reduced when stimulus rate was elevated to 20/s. These results suggest that VsEPs in response to linear accelerations are similar in their nature to VsEPs in response to angular acceleration impulses that were previously recorded. These VsEPs to linear accelerations are most likely initiated in the otolith organs. PMID- 9402895 TI - Activation of duration-sensitive auditory cortical fields in humans. AB - The influence of stimulus duration on auditory evoked potentials (AEPs) was examined for tones varying randomly in duration, location, and frequency in an auditory selective attention task. Stimulus duration effects were isolated as duration difference waves by subtracting AEPs to short duration tones from AEPs to longer duration tones of identical location, frequency and rise time. This analysis revealed that AEP components generally increased in amplitude and decreased in latency with increments in signal duration, with evidence of longer temporal integration times for lower frequency tones. Different temporal integration functions were seen for different N1 subcomponents. The results suggest that different auditory cortical areas have different temporal integration times, and that these functions vary as a function of tone frequency. PMID- 9402896 TI - Developmental changes in P1 and N1 central auditory responses elicited by consonant-vowel syllables. AB - Normal maturation and functioning of the central auditory system affects the development of speech perception and oral language capabilities. This study examined maturation of central auditory pathways as reflected by age-related changes in the P1/N1 components of the auditory evoked potential (AEP). A synthesized consonant-vowel syllable (ba) was used to elicit cortical AEPs in 86 normal children ranging in age from 6 to 15 years and ten normal adults. Distinct age-related changes were observed in the morphology of the AEP waveform. The adult response consists of a prominent negativity (N1) at about 100 ms, preceded by a smaller P1 component at about 50 ms. In contrast, the child response is characterized by a large P1 response at about 100 ms. This wave decreases significantly in latency and amplitude up to about 20 years of age. In children, P1 is followed by a broad negativity at about 200 ms which we term N1b. Many subjects (especially older children) also show an earlier negativity (N1a). Both N1a and N1b latencies decrease significantly with age. Amplitudes of N1a and N1b do not show significant age-related changes. All children have the N1b; however, the frequency of occurrence of N1a increases with age. Data indicate that the child P1 develops systematically into the adult response; however, the relationship of N1a and N1b to the adult N1 is unclear. These results indicate that maturational changes in the central auditory system are complex and extend well into the second decade of life. PMID- 9402897 TI - Reflection of working memory: ERP mnemonic effects. AB - The study of working memory often utilizes a delayed matching to sample paradigm (DMS). Typically in the matching condition, the test and sample stimuli are identical, raising the possible confound of retinotopic projections for the matching stimuli in contrast to the non-matching stimuli. In the present study, 65 healthy subjects performed a modified delayed matching to sample task while monitoring their ERP waveforms. The stimuli consisted of 60 different sample stimuli (S1) and 60 different test stimuli (S2). Half of the S2s were complementary to the sample stimuli (Fit), the other half of the S2s were not complementary (Nonfit). After S2, the subjects pressed one of the buttons to indicate whether the test stimulus fits the sample stimulus. Our statistical results indicated that the ERPs to sample stimuli differed from the ERPs to test stimuli from 200 ms poststimulus to the end of the recording epoch. The ERPs to fitting stimuli were significantly different from those to non-fitting stimuli from 200 to 400 ms poststimulus. The ERP patterns in the present study may reflect ERP mnemonic effect for working memory. Our results ruled out the retinotopic confound as a potential mediator variable, and are in agreement with other animal or human neurophysiological studies on memory. PMID- 9402898 TI - Use of nitrous oxide to dissociate the non-specific and specific components of the human auditory N1. AB - The components of the N1 are thought to be related to sensory functioning (Components 1 and 2) and arousal (Component 3). To provide direct evidence for the involvement of Component 3 in arousal, we hypothesized that it should be more sensitive to the anesthetic gas nitrous oxide (N2O) than Component 1. Using the technique of selective adaptation, 30 blocks of 5 tones were presented at 1 min intervals to 9 subjects who breathed air, 25% and 35% N2O. As hypothesized, the amplitude of Component 3 was significantly reduced in a dose-dependent manner by N2O, but the amplitude of Component 1 was not, although the latter showed some evidence of a decrease at 25% N2O. PMID- 9402899 TI - The new environment for health services research: private and public sector opportunities. PMID- 9402900 TI - Managed care: achieving the benefits, negating the harm. PMID- 9402901 TI - The determinants of dumping: a national study of economically motivated transfers involving mental health care. AB - OBJECTIVE: To examine the prevalence and determinants of economically motivated transfers (aka "dumping") from hospitals treating mental illness. DATA SOURCES: A composite data set constructed from three national random-sampled surveys conducted in 1988 and 1989: (1) of hospitals providing mental health care, (2) of community mental health centers, and (3) of psychiatrists. STUDY DESIGN: The study uses reports from administrators of community mental health centers (CMHCs) to assess the extent of patient dumping by hospitals. To assess the determinants of dumping, reported perceptions of dumping are regressed on variables describing the catchment area in terms of the proportion of for-profit hospitals, intensity of competition among hospitals, extent of utilization review, and capacity of the local treatment system, as well as competition among community mental health centers. To assess if dumping is motivated by factors distinct from those affecting other aspects of access, comparable regressions are estimated with ease of hospital admission as the dependent variables. PRINCIPAL FINDINGS: Economically motivated transfers of psychiatric patients were widespread in 1988: according to the reports of CMHC administrators, 64.7 percent of all hospitals providing inpatient mental health care engaged in transfers of this sort. The extent of dumping was higher in catchment areas with more competition among hospitals, more proprietary hospitals, and less inpatient capacity in the public sector. Dumping appeared to be more sensitive to capacity in the public sector but less sensitive to involvement by for-profit hospitals than were other measures of access to care. CONCLUSIONS: Economically motivated transfers of patients with mental illness were widespread in 1988 and likely have increased since that time, affecting the viability of the community mental health care system. PMID- 9402902 TI - Commentary: economic transfers, the changing face of a familiar problem. PMID- 9402903 TI - The cost and outcomes of community-based care for the seriously mentally ill. AB - OBJECTIVE: To examine the cost-effectiveness of community-based mental health care. DATA SOURCES/STUDY SETTING: Administrative data from Medicaid and the Massachusetts Department of Mental Health; primary data from 144 psychiatrically disabled adult Medicaid beneficiaries who lived in Boston, central Massachusetts, and western Massachusetts. STUDY DESIGN: A cross-sectional observational study compared the costs and outcomes of treatment in three different types of public mental health service systems. DATA COLLECTION/EXTRACTION METHODS: Beneficiaries, randomly sampled from outpatient mental health programs, were interviewed about their mental health status. All their acute treatment and long-term continuing care for the preceding year were abstracted from Medicaid and Department of Mental Health files. Costs were extracted from Medicaid paid claims and from Department of Mental Health contracts and other financial documents. PRINCIPAL FINDINGS: Clients in the region allocating a greater proportion of its Department of Mental Health budget to community support services used far fewer hospital days, resulting in lower per person treatment expenditures. Outcomes, however, were not significantly different from outcomes of clients in the other regions. For all regions, substance abuse comorbidity increased hospitalization and total treatment costs. An individual-level cost-effectiveness analysis identified western Massachusetts (community-based care) as significantly more cost effective than the other two regions. CONCLUSIONS: Systems with stronger community-based orientation are more cost effective. PMID- 9402904 TI - Cost-effectiveness of inpatient substance abuse treatment. AB - OBJECTIVE: To identify the characteristics of cost-effective inpatient substance abuse treatment programs. DATA SOURCES/STUDY SETTING: A survey of program directors and cost and discharge data for study of 38,863 patients treated in 98 Veterans Affairs treatment programs. STUDY DESIGN: We used random-effects regression to find the effect of program and patient characteristics on cost and readmission rates. A treatment was defined as successful if the patient was not readmitted for psychiatric or substance abuse care within six months. PRINCIPAL FINDINGS: Treatment was more expensive when the program was smaller, or had a longer intended length of stay (LOS) or a higher ratio of staff to patients. Readmission was less likely when the program was smaller or had longer intended LOS; the staff to patient ratio had no significant effect. The average treatment cost $3,754 with a 75.0% chance of being effective, a cost-effectiveness ratio of $5,007 per treatment success. A 28-day treatment program was $860 more costly and 3.3% more effective than a 21-day program, an incremental cost-effectiveness of $26,450 per treatment success. Patient characteristics did not affect readmission rates in the same way they affected costs. Patients with a history of prior treatment were more likely to be readmitted but their subsequent stays were less costly. CONCLUSIONS: A 21-day limit on intended LOS would increase the cost effectiveness of treatment programs. Consolidation of small programs would reduce cost, but would also reduce access to treatment. Reduction of the staff to patient ratio would increase the cost-effectiveness of the most intensively staffed programs. PMID- 9402905 TI - Performance contracting for substance abuse treatment. AB - OBJECTIVE: To describe an innovation in performance contracting for substance abuse services in the State of Maine and examine data on measured performance by providers before and after the innovation. DATA SOURCES AND COLLECTION: From the Maine Addiction Treatment System (MATS), an admission and discharge data set collected by the Maine Office of Substance Abuse (OSA). The MATS data for this study include information on clients of programs receiving public funding from October 1, 1989 through June 30, 1994. Additional data are drawn from the contracts between the state and providers, and from service delivery reports submitted to OSA. STUDY DESIGN: Client-level performance measures were calculated directly from MATS using OSA's formulas and standards, and then aggregated to the treatment program level. Multivariate regression analysis was done for each performance indicator as a dependent variable with performance contracting, time, extent of state funding, and provider characteristics as independent variables. PRINCIPAL FINDINGS: Performance contracting is positively related to better performance for effectiveness indicators overall. Individual effectiveness indicators that showed improvement include drug use indicators (abstinence and reduction in use) and social functioning indicators. In addition, performance contracting is associated with an increase in efficiency performance, defined as delivery of the contracted amount of service, for agencies that depend heavily on OSA for funding. Finally, performance contracting appears unrelated to the special populations indicators that measure services to target populations that OSA considers harder to treat. CONCLUSIONS: There is tentative evidence of a relationship between provider performance and the introduction of performance contracting. More definite conclusions await more detailed analyses of client level data. PMID- 9402906 TI - Rehabilitation costs: implications for prospective payment. AB - OBJECTIVE: To obtain information relevant to development of prospective payment for Medicare rehabilitation facilities (RFs) and skilled nursing facilities (SNFs): compares service utilization, length of stay (LOS), case mix, and resource consumption for Medicare patients receiving postacute institutional rehabilitation care. DATA SOURCES/STUDY SETTING: Longitudinal patient-level and related facility-level data on Medicare hip fracture (n = 513) and stroke (n = 483) patients admitted in 1991-1994 to a sample of 27 RFs and 65 SNFs in urban areas in 17 states. STUDY DESIGN: For each condition, two-group RF-SNF comparisons were made. Regression analysis was used to adjust RF-SNF differences in resource consumption per stay for patient condition (case mix) and other factors, since random assignment was not possible. DATA COLLECTION/EXTRACTION METHODS: Providers at each facility were trained to collect patient case-mix and service utilization information. Secondary data also were obtained. PRINCIPAL FINDINGS: RF patients had shorter LOS, fewer total nursing hours (but more skilled nursing hours), and more ancillary hours than SNF patients. After adjustment, ancillary resource consumption per stay remained substantially higher for RF than SNF patients, particularly for stroke. The adjusted nursing resource consumption differences were smaller than the ancillary differences and not statistically significant for hip fracture. Supplemental outcome findings suggested minimal differences for hip fracture patients but better outcomes for RF than SNF stroke patients. CONCLUSIONS: Much can be gained from an integrated approach to developing prospective payment for RFs and SNFs. In that context, consideration of condition-specific per-stay payment methods applicable to both settings appears warranted. PMID- 9402907 TI - Introduction: research on health care organizations and markets--the best and worst of times. PMID- 9402908 TI - Managed care research: moving beyond incremental thinking. PMID- 9402909 TI - Studying access to care in managed care environments. PMID- 9402910 TI - Managed care and chronic illness: health services research needs. PMID- 9402911 TI - Five priority areas for research on long-term care. PMID- 9402912 TI - Advances in the management of chronic depressive and anxiety disorders. Introduction. PMID- 9402913 TI - Panic disorder as a chronic illness. AB - Panic disorder is a chronic illness that waxes and wanes, and the prognosis is worse with comorbid agoraphobia, depression, or personality disorder. Both medication and cognitive-behavioral therapy have been found helpful in acute treatment trials of panic disorder. However, recent studies suggest that therapeutic gains are lost in many instances when treatment is stopped after short-term medication or cognitive-behavioral therapy. Thus, maintenance treatment appears necessary for many panic patients. Patients appear relatively stable during medication maintenance, but studies of maintenance psychosocial treatment for panic disorder have not yet been reported. Whether combined medication and psychosocial treatment lead to more durable effects after treatment discontinuation than are seen with individual treatments also remains to be determined. PMID- 9402914 TI - Treatment strategies for chronic and refractory obsessive-compulsive disorder. AB - Obsessive-compulsive disorder (OCD), despite our increasing understanding of its causes and its effective treatment, remains a chronic and underdiagnosed disorder. Both treatment with SSRIs and the behavioral treatment strategy of exposure with response prevention have been proved by clinical trials to be effective and safe in treating OCD; however, even these treatments sometimes elicit only moderate patient response, and some OCD patients do not respond to them at all. Preliminary data suggest that OCD has lifelong persistence and that discontinuation of pharmacotherapy often leads to relapse. Nonetheless, further prospective, controlled, maintenance studies of OCD are needed to determine factors in and predictors of recovery, remission, and relapse. Finally, new procedures for treatment-refractory patients are needed; neurosurgical and new pharmacologic approaches have shown promise in treating these patients and should be studied in controlled trials. PMID- 9402915 TI - Strategies and tactics in the treatment of chronic depression. AB - Chronic depressions include major depressive disorder, recurrent, without full interepisode recovery; major depressive disorder, currently in a chronic (i.e., > or = 2 years) episode; double depression; dysthymic disorder; and those depressive disorders--not otherwise specified (NOS)--that are persistent or predictably recurrent with substantial disability. Strategic treatment decisions include (1) whether to treat with medication, psychotherapy, ECT, or other methods; (2) selection among specific agents with long-term efficacy and tolerability (preferably established by randomized controlled trials); (3) selection of the next treatment should the initial treatment fail or be found intolerable; and (4) deciding whether to provide maintenance treatment. Tactical decisions are those needed to optimally implement the strategies selected; for example, (1) how to optimally dose, (2) how long to continue an acute phase treatment trial, (3) how to measure outcome, and (4) how to identify and manage subsequent symptomatic breakthroughs or side effects (which may also require revisions in the initial strategies). Some antidepressant medications evidence efficacy and safety in acute phase treatment of the chronically depressed, but continuation and maintenance phase treatments for these patients are less well investigated and deserve further study. The clinical implications of what is known to date for managing these patients are discussed. PMID- 9402916 TI - When at first you don't succeed: sequential strategies for antidepressant nonresponders. AB - Now, more than ever before, a wealth of options exists for depressed patients who do not benefit from treatment with standard, first-line antidepressant agents. In this paper, alternate antidepressant strategies are reviewed within the context of a five-stage strategy, ranging from lesser to greater degrees of treatment resistance. The overall strategy recommended progresses from simpler (i.e., an alternate monotherapy) to more complex strategies (i.e., combination or augmentation regimens), with the nonselective monoamine oxidase inhibitors (+/- lithium salts) and electroconvulsive therapy typically reserved for treatment of Stages III and IV of resistance, respectively. Psychotherapeutic management also is an important ingredient in the ongoing treatment of these patients, particularly to counteract the demoralization and frustration that understandably accompany the failure to respond to so many treatments. PMID- 9402917 TI - Psychotherapeutic approaches to the treatment of anxiety and depressive disorders. AB - Psychotherapy, both alone and in combination with pharmacotherapy, is one of the most prevalent treatments for depression and anxiety. Research data are sparse, but there is ample evidence that several psychotherapies are effective for acute affective and panic disorders. The best data are for interpersonal and cognitive behavioral therapies, with only early reports on the more common psychodynamic psychotherapies. There has been less study of more chronic disorders, but once again the suggestion is that appropriate psychotherapy is effective. Treatment should be active and focused on the patient's symptoms and current problems, not on character pathology or developmental psychodynamics. PMID- 9402918 TI - Controlled release vaccines. PMID- 9402919 TI - Resistance to beta-lactam antibiotics in Bacteroides spp. AB - Bacteroides spp., particularly B. fragilis, are well-recognised bacterial pathogens. Production of the typical beta-lactamases of Bacteroides restricts the therapeutic use of beta-lactam agents mainly to the beta-lactamase inhibitor combinations and carbapenems. These compounds have the advantage of broad spectrum activity and the ability to combat polymicrobial infections. Resistance of Bacteroides spp. to beta-lactam antibiotics appears to be increasing, largely because of an overall increase in beta-lactamase activity. There has been a rise in the prevalence of isolates showing high-level production of typical Bacteroides beta-lactamases and an increase in reports other potent beta lactamase types. In the case of B. fragilis, metallo-enzymes are a particular threat to current therapeutic practice, as they are not inhibited by common beta lactamase inhibitors and are able to hydrolyse carbapenems. The presence of permeability barriers may confer low-level beta-lactam resistance and supplement the effect of beta-lactamase activity. There are also sporadic reports of loss of beta-lactam activity because of reduced affinity of the penicillin-binding proteins. PMID- 9402920 TI - The isolation and cloning of chromosomal DNA specific for a clonal population of Staphylococcus aureus by subtractive hybridisation. AB - Subtractive hybridisation was used to screen for and identify conserved DNA sequences associated with clinical, clonal populations of Staphylococcus aureus. DNA from S. aureus strain ISP8 (a clinical isolate) was digested with TaqI and hybridised with randomly fragmented pooled DNA from 10 non-clinical (community) isolates of S. aureus. The mixture of DNA fragments was then ligated to AccI digested and dephosphorylated pTZ19U. One recombinant plasmid (pWASA) was identified as containing specific DNA from strain ISP8; DNA sequencing revealed a 40-bp TaqI DNA fragment (WASA). PCR amplification and hybridisation analysis, with pWASA as a probe, showed that 84% of clinical isolates from a clonal line present in hospitals in major eastern Australian cities carried sequences homologous to WASA, compared with only 10% of community isolates. The isolates that hybridised were closely related by RFLP analysis to strain ISP8. The plasmid pWASA was used to identify, isolate and clone a 3.5-kb DraI fragment (DSA) from strain ISP8 total DNA. Sequence analysis of the DSA fragment identified two open reading frames (ORFs) of 2475 and 576 bp, respectively; the larger ORF1 contained a series of six tandem repeats, each consisting of 384 nucleotides. The nucleotide sequence of the repeats was 96% identical, and no significant sequence homologies to previously described protein sequences were identified. However, tandem repeats of amino acids are structural motifs characteristic of a number of gram-positive surface proteins, and are thought to play a role in generating genetic and phenotypic diversity in these and other bacteria. PMID- 9402921 TI - Streptococcus agalactiae beta gene and gene product variations. AB - Streptococcus agalactiae (group B streptococci; GBS) are serotyped on the basis of the capsular polysaccharide antigens and subtyped on the basis of the strain variable and surface-localised c proteins c alpha, c beta, and R proteins. This study compared c beta protein detection and the polymerase chain reaction (PCR) for beta gene detection, by examining 50 clinical GBS strains. The c beta protein was detected by antibody-based immunofluorescence in a GBS whole-cell assay and Western blotting by probing with the anti-c beta antibody or human IgA. Absorption experiments were performed to test for surface-anchoring of c beta; and bacterial supernates were examined to test for c beta production. Primers for the PCR target regions resulted in a 620-bp product that included beta gene encoding IgA-binding domains. The results demonstrated four categories of GBS with respect to the beta gene and the c beta protein: (1) strains (16 of 50) that harboured the beta gene and regularly expressed normal surface-localised c beta with a M(r) of 120 kDa; (2) strains (5 of 50) that harboured the gene but did not express the protein; (3) strains (2 of 50) that harboured the gene but expressed a c beta that was not surface-localised and had reduced M(r); (4) strains (27 of 50) without beta gene and c beta expression. One strain amongst the third group generated a PCR product of 1330 bp. These results demonstrate considerable strain variability of the beta gene of GBS and of its product the c beta protein. PMID- 9402922 TI - Screening of TnphoA mutants of Vibrio cholerae O139 for identification of antigens involved in colonisation. AB - A new serogroup of Vibrio cholerae non-O1, designated as O139, has emerged causing cholera-like disease among adults. Laboratory and field studies clearly show that there is no cross-protection between O1 and O139 pathogenic strains. Since colonisation of the intestine is a most important step in the pathogenesis of cholera caused by O1 strains and colonising antigens are known to be protective, investigation of the colonising antigens of O139 strain was initiated. By TnphoA mutagenesis, mutants were generated with insertions in the genome encoding membrane spanning or secretory proteins. Screening of the mutants for adherence to rabbit intestinal surface and colonisation in 5-day-old mice resulted in the identification of mutant clones, which were less adhesive than was the wild-type parent strain and which could not efficiently colonise the gut. Such non-colonising strains were attenuated in virulence. Analysis of the proteins by SDS-PAGE revealed that the non-colonising mutants did not express a 40-kDa outer-membrane protein. PMID- 9402923 TI - Structural studies of the surface projections of Chlamydia trachomatis by electron microscopy. AB - Rod-like projections on the surface of Chlamydia trachomatis have been studied by a combination of computer image analysis and electron microscopy. The rods, c. 60 80 A in diameter and c. 500 A in length, were found on the surface of prokaryocells of C. trachomatis inserted in the cytoplasmic membrane through a ring-like structure in the outer membrane. The rod-like structures were found at all stages of the life cycle, even in very small elementary bodies (EBs) of C. trachomatis and in vesicles < 0.2 micron. Computer image analysis of isolated rods indicated that they comprise helically arranged subunits with a periodicity of c. 50 A. From their localisation and distribution, these structures may be related to the proliferation, or to the infectivity, of chlamydiae. PMID- 9402924 TI - An epidemiological study of Serratia marcescens isolates from nosocomial infections by enzyme electrophoresis. AB - Serratia marcescens isolates from 164 patients with suspected nosocomial infection in several hospitals in the greater Paris region were investigated by analysis of the electrophoretically demonstrable allelic variations of gene loci coding for five esterases and five other enzymes. All the loci were polymorphic and the mean number of alleles per locus was 6.1. A total of 72 distinctive electrophoretic types (ETs) representing multilocus genotypes was distinguished. The isolates were divided into two groups according to their resistance to antibiotics: 82 multiresistant isolates (MRI) and 82 relatively susceptible isolates (RSI). Seventy-two MRI (88%) were in four genetically related ETs: ET1, ET2, ET8 and ET9; ET1 was found in 48 isolates, whereas the remaining MRI were in 10 ETs, and all RSI in 61 ETs. Three ETs contained both MRI and RSI. The mean coefficients of genetic diversity for the 10 enzyme loci among ETs and isolates were smaller for MRI than for RSI, while the modal ET of MRI resembled that of RSI. The epidemiological significance of isolates varied according to their ET. Thus, isolates belonging to ET1, ET2 and ET8 were responsible for outbreaks or for sporadic infections, whereas isolates of other ETs were responsible for only sporadic infections. The temporal distribution of ET1 isolates among hospitals identified seven outbreaks in seven clinical departments. PMID- 9402925 TI - Enhancement of the specific mucosal IgA response in vivo by interleukin-5 expressed by an attenuated strain of Salmonella serotype Dublin. AB - It has been shown that cytokines have potential as therapeutic adjuvants in vaccination. Interleukin-5 (IL-5) is a cytokine that regulates antibody production, in particular enhancing IgA production by activated mucosal B cells. This study examined the expression of a cloned cytokine gene encoding murine IL-5 (mIL-5) by an attenuated aroA strain (SL5631) of Salmonella serotype Dublin. The resulting strain, SL5631(pTRXFUS-mIL-5), expressed mIL-5 as a fusion with thioredoxin as demonstrated by immunological and biological assays. When strain SL5631(pTRXFUS-mIL-5) was used as a live vaccine in BALB/c mice, it colonised and multiplied at higher levels in spleens and livers than the strain carrying the empty plasmid. A reduction in invasiveness of SL5631(pTRXFUS-mIL-5) was observed in in-vitro invasion assays. Enhanced IgA response against salmonella LPS in mucosal secretions and enhanced IgA and IgG responses were detected by ELISA and ELISPOT methods in sera of mice immunised with the strain expressing mIL-5. Results with IL-5-deficient mice showed that the enhanced IgA response was due to Salmonella-expressed mIL-5 rather than endogenous mIL-5. PMID- 9402926 TI - Low sensitivity of counter-current immuno-electrophoresis for serodiagnosis of typhoid fever. AB - Counter-current immuno-electrophoresis was evaluated as a diagnostic test for the serodiagnosis of typhoid fever with somatic (O), flagellar (H) and capsular polysaccharide (Vi) antigens of Salmonella typhi on the sera of patients who were blood culture positive (confirmed typhoid cases) or had high Widal agglutination titres, > or = 320, (presumptive typhoid cases). Of the 37 sera from confirmed cases, 30% showed positivity with O antigen, 24% with H antigens and 51% with Vi antigen. In patients with a presumptive diagnosis, 45% were positive for O antibody, 27% for flagellar antibody and 52% for Vi antibody. When all three antigens were combined the reactivity to any of the antigens was found to be 59% in confirmed typhoid cases, 79% in presumptive typhoid cases and 93% in patients who were simultaneously positive by blood culture and Widal agglutination. However, none of the sera from 45 controls gave a positive precipitation reaction with any of the antigens. It is concluded that counter-current immuno electrophoresis is a rapid test with low sensitivity and high specificity with Vi antigen, a panel of antigens being most effective, and is, therefore, recommended for rapid diagnosis of typhoid fever. PMID- 9402927 TI - PCR identification of Trichophyton mentagrophytes var. interdigitale and T. mentagrophytes var. mentagrophytes dermatophytes with a random primer. AB - Dermatophytes are a group of keratinophilic fungi falling within the genera of Epidermophyton, Microsporum and Trichophyton. The genus Trichophyton is particularly important and complex; it comprises at least 15 recognised species. In addition, there are several different variants in the species T. mentagrophytes, which occur both in man and animals. The current methods of determining T. mentagrophytes varieties may require several different culture media and time-consuming procedures, as well as specialist skills. This study used a random primer, 5'-GAGCCCGACT-3', in the arbitrarily primed polymerase chain reaction (AP-PCR) and showed that the two common T. mentagrophytes varieties (var. interdigitale and var. mentagrophytes) can be clearly identified on the basis of their characteristic DNA band patterns. The relative reproducibility, ease of use and precision of this method make the AP-PCR a valuable tool in the laboratory diagnosis of human dermatophytosis. PMID- 9402928 TI - Drugs for HIV infection. PMID- 9402929 TI - The age-associated alterations in late diastolic function in mice are improved by caloric restriction. AB - Caloric restriction reduces the magnitude of many age-related changes in rodents. Cardiac function is altered with senescence in mice, rats, and healthy humans. We examined the effects of life-long caloric restriction on diastolic and systolic cardiac function in situ using Doppler techniques in ad libitum-fed 30- to 32 month-old (AL) and calorically restricted (CR) 32- to 35-month-old female B6D2-F1 hybrid mice. The heart weight to body weight ratio was similar in AL (5.74 +/- .24 mg/g) and CR (5.68 +/- .20 mg/g) mice. Two systolic functional parameters known to decrease with age in both humans and mice, peak aortic velocity and aortic acceleration, were unchanged by CR compared to AL. In contrast, diastolic function was altered by caloric restriction. Although left ventricular peak early filling velocity (E) was not different between CR and AL, peak atrial filling velocity (A) was 50% lower in CR compared to AL (p < .001). The ratio of early diastolic filling to atrial filling (E/A ratio) was 64% higher in the CR (2.74 +/ .31) than the AL (1.55 +/- .07; p = .004). The fraction of ventricular filling due to atrial systole, the atrial filling fraction, was also reduced in CR (.21 +/- .04) compared to AL (.36 +/- .02; p = .007). These changes occurred in CR without alteration in E deceleration time, which is consistent with improved diastolic function in CR. Through mechanisms that remain unknown, lifelong caloric restriction may prevent the age-related impairments in late diastolic function but does not alter the impairments in systolic or early diastolic cardiac function. PMID- 9402930 TI - The effects of age and dietary restriction on metabotropic glutamate receptors in C57B1 mice. AB - This study examined the effects of aging and dietary restriction on metabotropic type 1 (met1; high quisqualate affinity) and type 2 (met2; low quisqualate affinity) binding sites for glutamate and the relationship between these binding sites and spatial memory. Quantitative autoradiography was performed on 3-, 10-, and 26-month-old mice. Ten- and 26-month-old mice were fed ad libitum or fed restricted diets. Met1 binding sites were not notably affected by age or diet. Binding to met2 sites was maintained during aging in the ad libitum-fed mice, but there were decreases in binding in the diet-restricted mice as compared to 3 month-old mice. Binding to met1 and met2 sites did not correlate significantly with performance in the Morris water maze. These results suggest that dietary restriction induces a decrease in one or more of the functions associated with met2 binding sites during the aging process. PMID- 9402931 TI - Effects of changes in calorie intake on intestinal nutrient uptake and transporter mRNA levels in aged mice. AB - In aged, chronically calorie-restricted (CR) mice, intestinal nutrient uptake is significantly higher than in same-age ad libitum controls. Can this chronic restriction-induced enhancement of uptake be reversed by ad libitum feeding? We addressed this question by switching 32-mo-old chronically CR mice to ad libitum feeding for 4 wk (CRAL). Intestinal transport rate and total intestinal absorptive capacity for D-sugars and several nonessential L-amino acids decreased significantly in CRAL mice. In contrast, switching CR mice to an ad libitum regimen for only 3 d had no effect on intestinal nutrient transport, indicating that the negative effects of ad libitum feeding require a duration longer than the 3-d lifetime of most enterocytes. Permeability of the intestinal mucosa to L glucose was independent of the switches in diet. Levels of the brushborder glucose transporter SGLT1, brushborder fructose transporter GLUT5, and basolateral sugar transporter GLUT2 mRNA as determined by reverse transcriptase polymerase chain reaction in 6-, 24-, and 32-mo-old mice were each apparently independent of caloric restriction and age. We conclude that the high rates of intestinal nutrient uptake exhibited by chronically CR mice can be reversed by ad libitum feeding of only 1 mo duration. These decreases in uptake were due mainly to specific decreases in transport per unit weight of intestine and not to nonspecific decreases in intestinal mass. Changes in rates of sugar uptake induced by chronic CR and age are apparently not accompanied by changes in steady state levels of mRNA coding for those transporters. PMID- 9402932 TI - Ethanol activates the interleukin-6 promoter in a human bone marrow stromal cell line. AB - Chronic ethanol consumption is associated with the development of osteoporosis. The pro-inflammatory cytokine interleukin-6 (IL-6) plays a role in the development of osteoporosis through stimulation of osteoclastic activity. We hypothesized that ethanol promotes osteoporosis, in part, by increasing IL-6 production in the bone microenvironment. Accordingly, we evaluated ethanol's effect on IL-6 production in the Saka human bone marrow stromal cell line and in the HOBIT human osteoblast-like cell line. We found that ethanol increased IL-6 protein levels in the culture supernatants from Saka, but not HOBIT, cells. In addition, we observed that ethanol increased steady-state IL-6 mRNA levels and activated an IL-6 promoter-driven reporter vector in Saka cells. We conclude that ethanol stimulates IL-6 expression in the Saka bone marrow stromal cell line by enhancing transcriptional activity of the IL-6 gene. Our findings support the contention that ethanol may contribute to the pathogenesis of osteoporosis, in part, by increasing IL-6 expression in the bone microenvironment. PMID- 9402933 TI - Aging stimulates fatty acid oxidation in rat colonocytes but does not influence the response to dietary fiber. AB - Metabolism was studied in colonocytes isolated from young (4 mo) and aged (24 mo) Fischer 344 rats. Animals were fed fiber-free, low-fiber (5% cellulose), or high fiber (oat bran or NIH 31 stock) diets. Colonocytes isolated from aged animals oxidized both short- and long-chain fatty acids at significantly higher rates than did colonocytes isolated from young animals. No differences between the young and aged were noted for the oxidation to CO2 of glucose and glutamine or for flux of glucose through glycolysis. Net adenosine triphosphate (ATP) production by colonocytes was calculated to be 20% higher for the aged than for the young, although the relative contribution of substrates to net ATP production from exogenous substrates was similar for the young and aged (45-50% from butyrate, 20-25% from glucose, and 30% from other substrates including acetate, propionate, palmitate, and glutamine). Substrate oxidation was generally higher in colonocytes from the oat bran (17% total dietary fiber, highly soluble fiber) versus fiber-free diet. PMID- 9402934 TI - Cellular and molecular biomarkers indicate precocious in vitro senescence in fibroblasts from SAMP6 mice. Evidence supporting a murine model of premature senescence and osteopenia. AB - A variety of short-lived mouse strains (SAMP strains) and control strains of less abbreviated life span (SAMR strains) have been proposed as murine models of accelerated senescence. Each SAMP strain, in addition to displaying "progeroid" traits of accelerated aging, exhibits a singular age-related pathology. The application of this animal model to the study of normal aging processes has been and remains controversial. Therefore, we have undertaken a study of dermal fibroblasts derived from the short-lived SAMP6 strain, which shows early-onset and progressive osteopenia. We have investigated cellular and molecular characteristics that are associated with in vitro aging of normal human fibroblasts, and which are exacerbated in fibroblasts from patients with Werner syndrome, a human model of premature senescence. We found that SAMP6 dermal fibroblasts, relative to SAMR1 and C57BL/6 controls, exhibit characteristics of premature or accelerated cellular senescence with regard to in vitro life span, initial growth rate, and patterns of gene expression. PMID- 9402936 TI - Geriatrics and medical oncology: finding the common ground. PMID- 9402935 TI - An evaluation of the length-tension relationship in elderly human plantarflexor muscles. AB - The purpose of this study was to determine the effect of aging on the muscle length-tension relationship in the plantarflexor muscles of 10 subjects aged 20 30 yr (Mean = 23; 5 males, 5 females), 10 subjects aged 60-80 yr (Mean = 72.3; 5 males, 5 females), and 10 subjects over 80 yr (Mean = 84.1, 5 males, 5 females). Isometric twitch properties, maximum voluntary strength, passive tension, and range of motion were measured at five different joint angles [20 degrees dorsiflexion (DF), 10 degrees DF, 0 degree, 10 degrees plantarflexion (PF), and 20 degrees PF]. Active (evoked and voluntary) and passive torque production were maximal when the ankle was rotated into the DF positions for all three age groups, whereas the lowest values were recorded when the ankle was rotated into 20 degrees PF. Males were stronger than females at all joint angles (p < .01). Also, young adults were stronger than both elderly adult groups (p < .01). These results illustrate that despite the considerable age-associated loss in both voluntary and evoked strength in the plantarflexors, the optimal angle for torque production remains the same for younger and older adults. PMID- 9402937 TI - Integration of aging and cancer research in geriatric medicine. PMID- 9402938 TI - Failure to thrive in old age: follow-up on a workshop. PMID- 9402939 TI - Determinants of bone mineral density in postmenopausal white Iowans. AB - BACKGROUND: Osteoporosis is a major health problem for older individuals. For women, development of osteoporosis is a function of the accretion of "peak" bone mass in the third decade, age at menopause, and rate of bone loss with aging. Low bone mineral density (BMD) is a major risk factor for osteoporosis and fracture. The purpose of this study was to identify life style, nutritional, medical, and genetic predictors of low BMD in postmenopausal Iowa women. METHODS: One hundred thirty-four postmenopausal White women ranging in age from 57 to 81 years were included in this case-control study. Bone mineral density was measured at the femoral neck, using dual photon X-ray absorptiometry (Hologic 2000 QDR). Sixty six women with BMD measurements below 0.68 g/cm2 (the bottom quartile of the BMD distribution in the population from which participants were recruited), and 68 women with values at or above 0.83 g/cm2 (the top quartile of the BMD distribution in the same population) were included. Information about environmental, nutritional, medical, and life style modifiers of BMD was obtained by written questionnaire and telephone interview. To assess familial factors that might influence BMD, we obtained a detailed family history for each participant. In addition, we tested the hypothesis that allelic variation at the Vitamin D receptor (VDR), and the type I collagen gene (COL1A1 and COL1A2) loci influence BMD. RESULTS: Weight, loss of height, age, and age at menopause were strong predictors of BMD in our population. After adjustment for these differences, we found no effect of genotype at the COL1A1, COL1A2, and VDR loci on BMD. CONCLUSIONS: Bone mineral density is a complex trait that is influenced by several different modifiers; in the present study, weight was the best predictor of postmenopausal BMD. While several studies suggest that VDR genotype is an important determinant of BMD, we did not find this association in our population, nor did we identify an association between allelic variation at the type I collagen gene loci and BMD. Identification of genes that determine body mass index may provide additional insight into risk factors for low BMD, and osteoporotic fractures. PMID- 9402940 TI - Comparison of alveolar bone loss, alveolar bone density and second metacarpal bone density, salivary and gingival crevicular fluid interleukin-6 concentrations in healthy premenopausal and postmenopausal women on estrogen therapy. AB - BACKGROUND: Osteoporosis is an age-related metabolic bone disease characterized by decreased mass and increased susceptibility to fracture. The literature suggests a relationship between oral bone loss and skeletal osteoporosis; however, most studies have produced conflicting results. The purpose of this study was to determine if a relationship exists among alveolar bone loss, alveolar bone density, second metacarpal density, salivary and gingival crevicular fluid interleukin 6 (IL-6), and IL-8 concentrations in premenopausal and postmenopausal healthy women receiving estrogen therapy. METHODS: Twenty eight healthy women (aged 23-78) were evaluated for this study. A vertical bitewing and hand radiographs were taken, and the subjects were evaluated for the presence of active periodontitis. The bitewing and hand radiographs were digitized, and measurements were made from the cemento-enamel junction to the alveolar crest from both arches. Bone density was evaluated in the maxillary and mandibular alveolar process and at the mid-shaft of the second metacarpal. Percent cortical area and the moment of inertia measurements were also determined. Stimulated whole saliva was collected for a 5-min period using a cube of paraffin as a stimulant and was analyzed for total protein by a colorimetric reaction and IL-6 and IL-8 by ELISA. RESULTS: The results of the study showed that postmenopausal women on estrogen therapy had more alveolar bone loss, more missing teeth, and reduced alveolar and second metacarpal bone density than premenopausal women. In addition, postmenopausal women on estrogen therapy had higher salivary IL-6 concentrations than premenopausal women. Alveolar bone densities were also strongly correlated to second metacarpal densities. CONCLUSIONS: The results of the study suggest that changes in alveolar bone density and levels of bone resorptive cytokines in saliva may be secondary to changes in menopausal status. These changes may predispose loss of alveolar bone with resultant loss of teeth. PMID- 9402941 TI - A single bout of concentric resistance exercise increases basal metabolic rate 48 hours after exercise in healthy 59-77-year-old men. AB - BACKGROUND: It has been shown that basal metabolic rate (BMR) decreases with age. The extent to which some of the decrease can be reversed by exercise in older men and women is unclear. Resistance exercise has been shown to significantly increase muscle mass in older individuals, and because muscle is a highly active metabolic tissue there is potential to increase BMR as a secondary outcome to the training adaptation. METHODS: Twelve healthy men aged 59-77 years performed single-leg knee extension exercise (right and left leg) and bench press lifts (16 sets, 10 reps/set with timed recovery between sets) at 75% of the individual's 3RM. Subjects only performed the concentric phase of the lift. BMR was measured on two separate occasions, once after a nonexercise control period and again 48 hrs after a bout of resistance exercise. RESULTS: BMR was significantly increased (p < .006) 48 hrs after exercise (EX) compared to control (CON) (284.0 +/- 34.0 vs 274.9 +/- 34.0 kJ/hr, respectively). Calculated over a 24-hour period, the energy expenditure corresponded to 1570 +/- 193 and 1627 +/- 193 kcal/24 hr (p < .0002) for the CON and EX measures, respectively. VO2 (L/min) was higher (p < .0002) 48 hrs after the EX bout compared to 48 hrs post-CON (0.232 +/- 0.03 vs 0.225 +/- 0.03 L/min, respectively). CONCLUSION: We conclude that in healthy 59 77-year-old men, an acute bout of resistance exercise causes a sustained increase in BMR that persists for up to 48 hours after exercise. PMID- 9402942 TI - Predictors of mobility decline: the Hong Kong old-old study. AB - BACKGROUND: Old age is often accompanied by functional decline and loss of autonomy. This longitudinal study examines the factors associated with mobility decline among a Chinese elderly cohort aged 70 years and above. METHODS: Analyses were carried out on data collected from 1,483 elderly subjects who were functionally mobile at baseline and survived the 18-month follow-up period. The outcome variable "mobility decline" was measured using the Barthel Activities of Daily Living Scale, which accesses subjects' ability to be independent in walking a distance of 50 meters and/or moving up and downstairs during the 18-month follow-up interview. RESULTS: Multivariate backward stepwise logistic regression analysis revealed that the following baseline characteristics were independently associated with mobility decline during the follow-up period: increasing age (OR 1.4, 95% CI 1.2-1.6 for every 5-year increase in age), no formal level of education (OR 1.9, 95% CI 1.0-3.9), no current practice of exercise (OR 2.1, 95% CI 1.4-3.1), symptoms of palpitation (OR 1.7, 95% CI 1.1-2.8), body mass index [weight (kg)/height (m)2] below 20 (OR 1.7, 95% CI 1.1-2.6), and slow gait velocity (OR 1.12, 95% CI 1.09-1.16 per second increase in gait time). There was also significant association between the experience of falls during follow-up and mobility decline (OR = 2.9, 95% CI = 1.9-4.5). CONCLUSION: Low body weight, lack of exercise, and falls during the follow-up period might serve as markers as to which subjects are at risk for mobility decline. PMID- 9402943 TI - Reproducibility of performance-based and self-reported measures of functional status. AB - BACKGROUND: The reproducibility of a performance-based and a self-reported measure of functional status was investigated, as well as the impact of age and cognitive function on the reproducibility. METHODS: Of a random sample of 114 men of the 1995 survey of the Zutphen Elderly Study, 105 men (aged 79.9 +/- 4.5 years) participated in a test-retest study. They filled out a questionnaire on disabilities and carried out performance tests twice, in a 2-week interval. Four performance tests were administered (standing balance, walking speed, chair stand, and external shoulder rotation), and a summary performance score was constructed. The number of self-reported disabilities in basic activities of daily living, mobility, and instrumental activities of daily living were assessed. Kappa statistics and Pearson correlation coefficients between test and retest measurements were computed for the total group and stratified by age and cognitive function. RESULTS: Three performance tests and the summary performance score had fair to good reproducibility (walking speed: Pearsons r = .90, chair stand: r = .82, shoulder rotation: kappa = .49, summary score: kappa = .52). Only the test for standing balance was poorly reproducible (kappa = .29). The self reported functional status was fairly to good reproducible (kappa = .63, r = .87). Self-reported functional status was significantly less reproducible in very old and cognitively impaired than in younger and nonimpaired individuals. CONCLUSIONS: In the elderly male subjects, performance tests and self-reported disabilities had moderate to good reproducibility, with the exception of the test for standing balance. In very old or cognitively impaired populations, self reported functional status may have a lower reproducibility. PMID- 9402944 TI - Management of verbally disruptive behaviors in nursing home residents. AB - BACKGROUND: Verbally disruptive behaviors (VDB) are verbal or vocal behaviors that are inappropriate to the circumstances in which they are manifested. These behaviors are a source of concern because they disturb persons around the older person and may be an indicator of distress. METHODS: Three interventions were tried and compared to a control no-intervention phase. The interventions were: (1) Presentation of a videotape of a family member talking to the older person, (2) in vivo social interaction, and (3) use of music. RESULTS: Thirty-two nursing home residents suffering from dementia and manifesting VDB were observed before, during, and after the interventions, and the duration of VDB was recorded. The behaviors decreased by 56% during the social interaction, 46% during the videotape, 31% during the music, and 16% during the no-intervention. CONCLUSIONS: The effects of the interventions were clinically and statistically significant, indicating the importance of providing stimulating activities and a richer environment to cognitively impaired nursing home residents. PMID- 9402945 TI - Primary hepatocyte culture as an experimental model for the evaluation of interactions between xenobiotics and drug-metabolizing enzymes. PMID- 9402946 TI - Preclinical evaluation of drug-drug interaction potential: present status of the application of primary human hepatocytes in the evaluation of cytochrome P450 induction. AB - Cytochrome P450 (CYP) inhibition and induction are the key mechanisms in drug drug interactions. Aside from clinical studies, primary human hepatocytes may represent the most appropriate experimental system for the evaluation of CYP induction in humans. A consensus of an international panel on the present status and future research directions in the application of primary human hepatocytes in the evaluation of CYP-induction is presented here. The following observations are concluded to be generally true: (1) Human hepatocytes isolated from both biopsy samples and transplantable livers are suitable for induction studies. (2) Hormonally-defined media can be used for the evaluation of CYP induction. (3) Isozyme-selective induction of CYP1A and 3A by known inducers are observed. (4) Reproducibility of induction could be improved by using hepatocytes plated as confluent cultures. (5) Induction could be observed for hepatocytes treated at 1 3 days after culturing. (6) Treatment duration of 2 days in general leads to near maximal induction. (7) In general, there is a good qualitative correlation between human hepatocyte results in vitro and clinical observations in vivo. (8) When the same inducers were evaluated in independent laboratories, similar data were generally observed. We conclude that primary human hepatocytes represent an appropriate model for mechanistic evaluation of CYP induction and as a screening tool for CYP induction potential of xenobiotics. A set of data acceptance criteria are proposed: (1) Positive response should be observed with concurrent positive control chemicals; (2) reproducible observation should be observed with multiple human donors; (3) for negative response, the doses used should not be cytotoxic; and (4) replicate treatment and/or multiple dose treatment should be performed to allow statistical analysis. Future studies should include the further development of on: (1) The inducibility of CYP isozymes other than CYP1A and 3A, and phase II enzymes; (2) further development of culturing condition to allow optimal gene expression; (3) evaluation of the involvement of nonparenchymal cells on CYP induction of parenchymal cells; (4) the and validation of quantitative approaches to extrapolate in vitro data to in vivo data; (5) evaluation of possible individual variations and potential genetic polymorphism in inducibility; (6) further definition of species differences in CYP induction; (7) development of a 'normal' human hepatocyte cell line for CYP induction studies; (8) improvement of cryopreservation procedure of human hepatocytes; (9) definition of the molecular mechanisms of CYP induction; and (10) evaluation of the induction of phase II metabolic pathways. PMID- 9402948 TI - Long-term primary cultures of adult human hepatocytes. AB - In this work we have investigated a system of long-term primary cultures of adult human hepatocytes which, in contrast to those previously described, has the advantage of requiring neither the use of additive cells as in co-cultures, nor of matrix component preparations like Matrigel or collagen sandwich. This system has been used previously for long-term cultures of hepatocytes from young baboon, and some modifications have been introduced here to take into account the specificity of adult human hepatocytes. In this system, hepatocytes are plated at confluence on collagen-coated dishes and cultured in a serum-free medium consisting of Williams'E supplemented with hormones and growth factors. Proteins secreted specifically by the liver, including albumin, alpha-1 antitrypsin, plasminogen, fibrinogen, lipoproteins ApoA1 and ApoB100, have been quantified in the extracellular medium as a function of time, either by immunoblot or ELISA. In addition, the expression and inducibility of CYP proteins of the CYP1, CYP2 and CYP3 families in response to their prototypical inducers including 2,3,7,8 tetrachlorodibenzo(p)dioxin and rifampicin, have been evaluated by immunoblot analysis of microsomes or cell lysates. Moreover, the oxidative metabolism of cyclosporin A, a monoxygenase activity depending on CYP3A4, has been monitored directly on the cultured cells by HPLC analysis of extracellular medium. Our results show that, under these culture conditions, adult human hepatocytes retain these phenotypical characteristics for at least 35 days. This system meets the requirements for use as a model for screening CYP protein inducers. PMID- 9402947 TI - Primary human hepatocytes as a tool for the evaluation of structure-activity relationship in cytochrome P450 induction potential of xenobiotics: evaluation of rifampin, rifapentine and rifabutin. AB - In our laboratory, primary human hepatocytes are being investigated as an in vitro experimental system for the evaluation of pharmacokinetic drug-drug interactions. Our study here represents the first reported study that directly compares the cytochrome P450 isozyme 3A (CYP3A) induction potential of three antimicrobials derived from rifamycin B, namely, rifampin, rifapentine and rifabutin. Two endpoints of CYP3A activity, testosterone 6 beta-hydroxylation and midazolam 1-hydroxylation have been used. Results obtained with hepatocytes from four different human donors show consistently that rifampin and rifapentine are potent inducers of CYP3A, while a significantly lower induction potential is observed for rifabutin. The relative induction potency of the three antimicrobials (rifampin > rifapentine >> rifabutin) is consistent with the available human in vivo data. For CYP1A measured as ethoxyresorufin O-deethylase activity, CYP2C8/9 measured as tolbutamide 4-hydroxylation activity, CYP2D6 measured as dextromethorphan O-demethylation, and AZT glucuronidation, there is either no effect or, where induction is found to be statistically significant in these other endpoints, the maximum induction values are consistently < 100% of the control. Our results suggest that CYP3A is the major CYP induced by these rifamycin B derivatives. These studies illustrate the application of human hepatocytes in the evaluation of the structure-activity relationships in CYP induction for this class of chemicals and as an in vitro screen for drug-drug interaction potential via CYP induction. PMID- 9402949 TI - Quantitative reverse transcriptase/PCR assay for the measurement of induction in cultured hepatocytes. AB - Hepatic enzyme induction is often evoked as the cause for a variety of effects in animal studies, e.g. hepatic hypertrophy and secondary thyroid neoplasms in rodents. In clinical practice, enzyme induction often enhances drug clearance and may result in reduced efficacy. For example, carbamazepine or rifampin treatment induces P450 3A in humans, and as a result, dramatically reduces the efficacy of midazolam or cyclosporine. Due to species differences in substrate specificity and the regulation of various drug-metabolizing enzymes, assessing enzyme induction in human tissues is a desirable goal. Since induction often occurs as a result of increased synthesis of mRNA coding for a particular enzyme, induction may be quantified by measuring specific mRNA levels. We describe an approach to quantifying mRNA levels using reverse transcriptase-polymerase chain reaction (RT PCR). This approach makes use of either radiolabeled PCR primers or fluorimetric quantification of product and does not require the synthesis of a competitor RNA or DNA molecule. Thus, Cyp2B1/2 mRNA can be shown to be induced 17-fold in the H4IIE rat hepatoma cell line. Likewise, Cyp3A and Cyp2A6 mRNAs can be shown to be induced in primary human hepatocytes cultured on collagen-coated plates and treated with rifampin for 72 h. By contrast, mRNA levels for Cyp1A1 and Cyp2E1 were not increased by rifampin treatment. This report demonstrates the potential of this approach for examining a number of mRNAs from drug-treated cultured cells, as a means of assessing metabolic enzyme induction. PMID- 9402950 TI - An evaluation of the CYP1A induction potential of pantoprazole in primary rat hepatocytes: a comparison with other proton pump inhibitors. AB - The ability of pantoprazole to affect the induction of cytochrome P450 (CYP) 1A subfamily was evaluated and compared with two other proton pump inhibitors, omeprazole and lansoprazole, in primary cultured hepatocytes from female Sprague Dawley rats. The hepatocytes were cultured for 2 days, followed by treatment for 2 days with the proton pump inhibitors at 2, 5 and 10 microM, concentrations that are similar to plasma concentrations found in rats in vivo. The CYP1A inducer 3 methylcholanthrene (at 1 microM) was also evaluated as a positive control. Induction potentials of these chemicals for CYP1A were determined by 7 ethoxyresorufin O-deethylase activity and isozyme contents. The results showed that for CYP1A induction, the rank ordering in induction potential was consistently lansoprazole > omeprazole > pantoprazole. The results are consistent with the existing rat in vivo data, i.e. pantoprazole has lower CYP1A induction potential than omeprazole and lansoprazole. PMID- 9402951 TI - Insulin effects on CYP2E1, 2B, 3A, and 4A expression in primary cultured rat hepatocytes. AB - Although hyperketonemia and/or altered growth hormone secretion caused by diabetes have been implicated in enhanced CYP2E1, 2B, 3A and 4A expression, the effect of insulin on hepatic P450 expression, in the absence of associated metabolic/hormonal alterations, remains unknown. Primary cultured rat hepatocytes have been shown (Zangar et al., Drug Metab. Dispos., 23:681, 1995) to express stable and inducible CYP2E1 mRNA and protein levels, and provide an excellent system for mechanistic examination of the effect of insulin on CYP2E1, 2B, 3A and 4A expression. Maintaining primary rat hepatocytes in culture in the absence of insulin for 48, 72, or 96 h increased CYP2E1 mRNA levels 5-, 11-, and 4-fold, respectively, relative to cells maintained in the presence of the standard concentration of 1 microM insulin. In contrast, CYP2B mRNA levels increased only approximately 2-fold in the absence of insulin, when compared with the presence of 1 microM insulin. CYP2E1 and 2B protein levels were increased 6.7- and 3.8 fold, respectively, in cells cultured for 96 h in the absence of insulin as compared with those cultured in medium containing 1 microM insulin. Concentration response studies revealed that decreasing the concentration of insulin below 10 nM (i.e. 1 nM, 0.1 nM, no insulin) increased CYP2E1 mRNA levels 4-, 7-, and 11 fold, respectively. In contrast, no such concentration-dependence was observed for CYP2B mRNA expression. As CYP3A and 4A expression is also elevated in diabetic rats, the effects of insulin on these P450s was also examined. CYP3A mRNA levels were unaltered and CYP4A mRNA levels were decreased marginally (approximately 50%) by the absence of insulin relative to levels in cells cultured in the presence of 1 microM insulin over 96 h in culture. The results of this study provide evidence that insulin itself, in the absence of other diabetes induced metabolic or hormonal alterations, affects CYP2E1 and 2B, but not CYP3A or 4A, expression in primary cultured rat hepatocytes. Furthermore, CYP2E1 expression is differentially regulated by insulin relative to CYP2B, 3A or 4A. This study also demonstrates that decreasing the concentration of insulin in the culture medium provides a method by which CYP2E1 levels can be increased in primary cultured hepatocytes to facilitate mechanistic studies on the regulation of CYP2E1 expression. PMID- 9402952 TI - Pig hepatocytes as an in vitro model to study the regulation of human CYP3A4: prediction of drug-drug interactions with 17 alpha-ethynylestradiol. AB - The objective of this study was to provide evidence of the validity of pig hepatocytes as a model to study the regulation of human CYP3A4 with special emphasis on drug-drug interactions. Thirteen different drugs were incubated with primary monolayer cultures of pig hepatocytes (n = 4). The study included both drugs reported to cause drug interactions in the clinic with 17 alpha ethynylestradiol (EE2), other drugs metabolized by CYP3A4, and drugs not reported to cause any problems. Effect of the drug exposure to pig hepatocytes was determined by immunodetection using a monoclonal human CYP3A4 antibody and measurement of 6 beta-hydroxylation of testosterone and 2-hydroxylation of 17 alpha-ethynylestradiol (EE2), both reactions known to be catalyzed by CYP3A4 in humans. Data were compared to data from human hepatocytes and to reported observations of drug-drug interactions in the clinic. The drugs known to be inducers of CYP3A4 in humans significantly increased a CYP isoform in pigs catalyzing 6 beta-hydroxylation of testosterone and 2-hydroxylation of EE2, whereas drugs not reported to have clinical interactions with EE2 had no or only marginal effect. Induction by the drugs known to be inducers of CYP3A4 increased with drug exposure time and the CYP3A4 activity, represented by testosterone 6 beta-hydroxylation, was highest at 72 h for the investigated induction periods (24, 48 and 72 h), except for dexamethasone where the effect peaked after 24 h. Induction of the 2-hydroxylation of EE2 correlated well with the increase in 6 beta-hydroxylation of testosterone (except for sulphinpyranzone) and the increase in the protein level of CYP3A detected by a monoclonal human CYP3A4 antibody, thus confirming the 2-hydroxylation of EE2 in pigs as being biotransformed by a CYP isoform presumably belonging to the CYP3A subfamily as in humans. In conclusion, these results indicate that pig hepatocytes may be a valuable model to mimic the regulation of human CYP3A4. PMID- 9402953 TI - Extrapolation of in vitro enzyme induction data to humans in vivo. AB - Enzyme induction generally increases the rate and extent of xenobiotic metabolism in vitro, but physiological constraints can dampen these effects in vivo. Biotransformation kinetics determined in hepatocytes in vitro can be extrapolated to whole animals based on the hepatocellularity of the liver, since the initial velocity of an enzyme-catalyzed reaction is directly proportional to the total enzyme present in the cell. The biotransformation kinetics of various xenobiotics determined with isolated hepatocytes in vitro have been shown to accurately predict pharmacokinetics in whole animals. Analysis of the kinetic data, using physiologically based pharmacokinetics, allows extrapolation of xenobiotic biotransformation across dose routes and species in a biologically realistic context. Several fold variations were observed in the bioactivation of the hepatotoxicant furan by isolated human hepatocytes, due to induction of cytochrome P450 2E1. Extrapolation of these data to humans in vivo showed that furan bioactivation was limited by hepatic blood flow delivery of the substrate. One important consequence of hepatic blood flow limitation is that the amount of metabolite formed in the liver is unaffected by increases in Vmax due to enzyme induction. Therefore, interindividual variations in cytochrome P450 2E1 among human populations would not affect the bioactivation of many rapidly metabolized hazardous chemical air pollutants. The hepatic blood flow limitation of biotransformation is also observed after oral bolus dosing of rapidly metabolized compounds. More slowly metabolized xenobiotics, such as therapeutic agents, are only partially limited by hepatic blood flow and other processes. PMID- 9402954 TI - An apoptosis-inducing genotoxin differentiates heterozygotic carriers for Werner helicase mutations from wild-type and homozygous mutants. AB - Immortalized B lymphocytes from Werner syndrome subjects are shown to be hypersensitive to 4-nitroquinoline-1-oxide (4NQO), supporting earlier work on T lymphocytes. We also show that B cell lines from clinically normal heterozygous carriers exhibit sensitivities to this genotoxic agent, which are intermediate to those of wild-type and homozygous mutants. 4NQO is shown to induce an apoptotic response. These data encourage research on DNA repair with such cell lines and raise the question of an enhanced sensitivity of the relatively prevalent heterozygous carriers to certain environmental genotoxic agents. PMID- 9402955 TI - Association of transferrin C2 allele with late-onset Alzheimer's disease. AB - Transferrin (Tf), an iron-transporting protein, has many variants, but C1 and C2 variants account for the majority of the population in all races. Since Tf is reported to be immunocytochemically detectable in senile plaques in Alzheimer's disease (AD), we have examined the Tf allele frequency among AD patients. The C2 allele frequency in late-onset AD patients is significantly higher than that in age-matched controls. Unexpectedly, the C2 allele frequency in AD patients homozygous for the ApoE epsilon 4 allele is markedly increased, i.e., it is twice as high as that in the remaining AD patients carrying zero or one copy of the epsilon 4 allele. PMID- 9402956 TI - Fine mapping of the gene for autosomal dominant juvenile-onset glaucoma with iridogoniodysgenesis in 6p25-tel. AB - Glaucoma is a group of ocular disorders leading to reduced visual capabilities and sometimes blindness. The biochemical defect is unknown but it is shown that reduced drainage of the aqueous humour from the anterior chamber may lead to increased intraocular pressure and gradual atrophy of the optic neurons. Families with various forms of autosomal dominant (AD) glaucoma have been linked to 1q21 31, 2cen-q13, 4q25-27, and 13q14 and autosomal recessive congenital glaucoma have been localized to chromosome 1p36 and 2p21. Recently, a locus for AD iridogoniodysgenesis anomaly (IGDA) was mapped to chromosome 6p25. This study refines the localization of IGDA to an approximately 6-cM interval between D6S1600 and D6S1617/D6S1713 at 6p25-tel, based on recombinations in affected individuals with AD juvenile-onset glaucoma and concomitant iridogoniodysgenesis. PMID- 9402957 TI - Evolutionary origins of two tightly linked mutations in arylsulfatase-A pseudodeficiency. AB - Deficient arylsulfatase A activity causes the neurodegenerative disease metachromatic leukodystrophy. However, some individuals with deficient enzyme activity appear clinically normal. This "pseudodeficiency" allele commonly found among many reported populations (frequency approximately 0.10) is associated with two A-->G transitions in cis in the arylsulfatase A gene causing the simultaneous loss of an N-glycosylation and a polyadenylation signal. To understand the evolutionary relationship between such common and tightly linked mutations, we studied 400 individuals in the African, European, Indian and East Asian populations and found none carrying the polyadenylation mutation alone. However, the N-glycosylation mutation could occur independently. Its frequency varied from 0.01 in Indians, 0.06 in Europeans, 0.21 in East Asians to 0.32 in Africans. The frequencies of both mutations occurring together ranged from almost non-existent in the Africans and East Asians, to 0.075 in the Europeans and 0.125 in the Indians. These frequencies were significantly different among populations. Haplotype analysis among homozygous pseudodeficiency individuals and eight multi generation families with six polymorphic enzymes showed that, of the five haplotypes found in the general population, only one was linked to the double mutations. Alleles among the four populations with only the N-glycosylation mutation also supported linkage to the same haplotype except in some Europeans whose alleles were discordant. These results are consistent with the hypothesis that the N-glycosylation mutation may be a recurrent event among the Europeans but first occurred in an ancestral allele before the emergence of modern Homo sapiens from Africa at approximately 100,000-200,000 years ago. Subsequently, the polyadenylation mutation occurred in this ancient allele with the N-glycosylation mutation, an event that likely took place after the divergence between the European and East Asian lineages. PMID- 9402958 TI - Evidence for convergent evolution of A and B blood group antigens in primates. AB - To determine whether convergent or trans-specific evolution is responsible for the persistence of the ABO polymorphism in apes, we have sequenced segments of introns 5 and 6 of the ABO gene. Four substitutions and one insertion or deletion group human A, B, and O alleles together, separate from their chimpanzee A and gorilla B counterparts. No shared substitutions support a trans-species mode of evolution for any of the alleles examined. We conclude that the A and B antigens of the chimpanzee and gorilla, respectively, have arisen by convergent evolution. Phylogenetic analysis suggests that the human A and B alleles are ancient, having diverged at least 3 million years ago. These alleles must have therefore been trans-specifically inherited within the genus Homo. PMID- 9402959 TI - A novel source of highly specific chromosome painting probes for human karyotype analysis derived from primate homologues. AB - We have established a series of highly specific painting probes for human acrocentric chromosomes. These chromosomes are involved in the formation of the nucleolar organizer region (NOR) and show DNA sequence homologies within their pericentric heterochromatin. To date, these chromosomes have shown considerable cross hybridization in chromosome painting experiments. Our probe set has been established from primate homologues that are not involved in the NOR in that particular species or from species in which highly repetitive sequences have undergone rapid sequence divergence. The new painting probes should be of particular value for automated microscopy, for which highly specific signals are required as they are recorded at low magnification, e.g. when scoring chromosome 21 domains in interphase nuclei. PMID- 9402960 TI - Gene structure and allelic expression assay of the human GLI3 gene. AB - The GLI3 gene encodes a putative zinc finger transcription factor that is important in early vertebrate development. Haploinsufficiency of this gene has been associated with the Greig cephalopolysyndactyly syndrome and truncation mutations cause Pallister-Hall syndrome. In the course of studies to determine the etiology of Pallister-Hall syndrome, we required knowledge of the fine structure of GLI3 to perform detailed genetic and physical mapping and mutation screening of this gene. The coding region of GLI3 is composed of 14 exons, including a large exon of more than 2500 bp. In addition, the gene contains two intragenic dinucleotide repeats, and four single-base pair polymorphisms in the coding region. We have used these coding region polymorphisms to design an allele specific expression study that will be useful for studying patients with Greig cephalopolysyndactyly syndrome. In addition, GLI3 should be considered a candidate gene for related developmental anomalies of humans. Such hypotheses will be more readily addressed with the availability of the fine structure of the gene and the allele-expression assay. PMID- 9402961 TI - ACP1 and human adaptability. 2. Association with season of conception. AB - We have studied the pattern of association between the season of conception and cytosolic low molecular weight phosphotyrosine phosphatase (ACP1) genetic polymorphism in 329 consecutively newborn infants from the population of Penne and 361 consecutively newborn infants from the population of Rome. In addition, 329 mothers were studied in the population of Penne. A concordant, highly significant association was observed in the two populations between ACP1 parameters and the season of conception of newborn infants. The total activity of ACP1 shows a minimum in infants conceived in January-February and a maximum in those conceived at the end of the solar year. Analysis of the joint mother newborn ACP1 distribution in relation to the season of fertilisation has shown that among mothers carrying ACP1*A (the allele showing the lowest activity), the proportion of newborns carrying this allele is higher in those conceived in the first months of the year than in those conceived subsequently. Since ACP1 probably functions as a phosphotyrosine phosphatase and as a flavin mononucleotide phosphatase, low activity could enhance the metabolic rate and would be advantageous in a cold environment. The cycle of variation of ACP1 in infants follows the cycle of solar illumination. It is possible that individuals who have a genetic background allowing them to adapt easily and readily to seasonal demand are more successful in reproducing themselves. The population of zygotes conceived in a given season would therefore reproduce the pattern of gene combination more fit for that season. PMID- 9402962 TI - A new mutation in the type II hair cortex keratin hHb1 involved in the inherited hair disorder monilethrix. AB - Monilethrix is a rare dominant hair disease characterized by beaded or moniliform hair which results from the periodic thinning of the hair shaft and shows a high propensity to excess weathering and fracturing. Several cases of monilethrix have been linked to the type II keratin gene cluster on chromosome 12q13 and causative heterozygous mutations of a highly conserved glutamic acid residue (Glu 410 Lys and Glu 410 Asp) in the helix termination motif of the type II hair keratin hHb6 have recently been identified in monilethrix patients of two unrelated families. In the present study, we have investigated two further unrelated monilethrix families as well as a single case. Affected members of one family and the single patient exhibited the prevalent hHb6 Glu 410 Lys mutation. In the second family, we identified in affected individuals a lysine substitution of the corresponding glutamic acid residue, Glu 403, in the type II hair keratin hHb1, suggesting that this site represents a mutational hotspot in these highly related type II hair keratins. Both hHb1 and hHb6 are largely coexpressed in cortical trichocytes of the hair shaft. This indicates that monilethrix is a disease of the hair cortex. PMID- 9402963 TI - Phosphorylase-kinase-deficient liver glycogenosis with an unusual biochemical phenotype in blood cells associated with a missense mutation in the beta subunit gene (PHKB). AB - We have identified mutations in the phosphorylase kinase (Phk) beta subunit gene in a male patient with liver glycogenosis caused by Phk deficiency. The patient's DNA has been analyzed for mutations in the genes encoding the alpha L, beta, and gamma TL subunits of Phk, all of which can be responsible for liver glycogenosis, by a strategy primarily based on reverse transcription/polymerase chain reaction of blood RNA and complemented by analysis of genomic DNA. His alpha L and gamma TL coding sequences are normal, whereas he is compound-heterozygous for two mutations in the beta subunit gene, PHKB. The first is a splice-site mutation (IVS4 [-2A-->G]) causing the reading-frame-disrupting deletion of exon 5 in the mRNA from this allele. The second is an Ala117Pro missense mutation, also in exon 5. This is the first missense mutation identified in PHKB, as opposed to nine translation-terminating mutations described to date. It offers an explanation for the unique biochemical phenotype of this patient. In his leukocytes, low Phk activity is measured when tested with the endogenous liver isoform of phosphorylase as the protein substrate, but normal activity is observed when tested with muscle phosphorylase added in vitro. In contrast, Phk activity in his erythrocytes is low with both substrates. The missense mutation may selectively impair the interaction of Phk with one isoform of its substrate protein and may destabilize the enzyme in a cell-type-specific way. This phenotype shares some aspects with X-linked liver glycogenosis subtype 2 (XLG2), a variant of liver Phk deficiency arising from missense mutations in the alpha L subunit gene (PHKA2), but differs from XLG2 in other respects. The present case demonstrates that mutations in Phk genes other than PHKA2 can also be associated with untypically high activity in certain blood cell types. Moreover, it emphasizes that missense mutations in Phk may cause unusual patterns of tissue involvement that would not be predicted a priori from the tissue specificity of expression of the mutated gene sequences. PMID- 9402964 TI - Inv(10)(p11.2q21.2), a variant chromosome. AB - We present 33 families in which a pericentric inversion of chromosome 10 is segregating. In addition, we summarise the data on 32 families in which an apparently identical inv(10) has been reported in the literature. Ascertainment was through prenatal diagnosis or with a normal phenotype in 21/33 families. In the other 12 families, probands were ascertained through a wide variety of referral reasons but in all but one case (a stillbirth), studies of the family showed that the reason for referral was unrelated to the chromosome abnormality. There has been, to our knowledge, no recorded instance of a recombinant chromosome 10 arising from this inversion and no excess of infertility or spontaneous abortion among carriers of either sex. We propose that inv(10)(p11.2q21.2) can be regarded as a variant analogous to the pericentric inversion of chromosome 2(p11q13). We conclude that prenatal chromosome analysis is not justified for inv(10) carriers. In addition, family investigation of carrier status is not warranted in view of the unnecessary concern this may cause parents and other family members. PMID- 9402965 TI - RB1 deletion in gonadoblastoma in an XY female. AB - Cytogenetic studies of normal and tumor cells in a patient with gonadal dysgenesis and bilateral gonadoblastoma were performed. The karyotype was 46,XY in peripheral blood lymphocytes and skin fibroblasts. The conserved region of the SRY gene was detected by polymerase chain reaction amplification. Sequencing of this region did not reveal any alterations. A 46,XY chromosome constitution was observed in the right gonadoblastoma, but a partial deletion of chromosome 13 was present in the left tumor. This deletion included band 13q14, where the retinoblastoma gene is mapped. The study of the polymorphism of the variable number of tandem repeats region in intron 17 of the RB1 locus disclosed loss of heterozygosity in both the left tumor, which showed the deletion of chromosome 13, and in the right tumor, where no chromosome alterations of chromosome 13 were detected. In situ hybridization covering 130 kb of RB1 showed that a partial deletion of one of the RB1 alleles had occurred in the right tumor. Since the deletions affected different alleles in each tumor, independent events must have been involved in the development of the tumors. These findings point toward a significant role of RB1 in the development of gonadoblastoma. PMID- 9402966 TI - No founder effect detected in Jewish Ashkenazi patients with fragile-X syndrome. AB - Several studies on small homogenous populations suggested that fragile-X syndrome originated from a limited number of founder chromosomes. The Israeli Jewish population could serve as an adequate model for tracing a founder effect due to the unique ethnic makeup and traditional lifestyle. Furthermore, a common haplotype for Jewish Tunisian fragile X patients was recently reported. To test for a similar occurrence in the Jewish Ashkenazi population, we performed haplotype analysis of 23 fragile-X patients and 28 normal chromosomes, all Jewish Ashkenazi, using microsatellite markers within and flanking the FMR-1 gene: FRAXAC1, FRAXAC2, and DXS548. The combined triple-marker analysis identified a wide range of diverse haplotypes in patients and controls, with no distinct haplotype prevalent in the patient group. Our data suggest that no common ancestral X chromosome is associated with the fragile-X syndrome in the Israeli Jewish Ashkenazi patient population studied. These findings are in contrast to other reports on founder effect associated with fragile-X syndrome in distinct European as well as Jewish Tunisian populations. On this basis, a more complex mechanism for the development of fragile-X syndrome in the Jewish Ashkenazi population should be considered. PMID- 9402967 TI - Identification of an amplified gene cluster in glioma including two novel amplified genes isolated by exon trapping. AB - Gene amplification, which occurs in more than 50% of malignant gliomas, is considered to play a pivotal role in tumorigenesis. There are, however, few studies aimed toward the isolation of novel genes from amplified sequences. Previously, we reported amplification of the protooncogene MET (hepatocyte growth factor receptor; 7q31) in more than 20% of glioblastomas. For an approximate size estimation of the amplification unit we analyzed three glioblastomas all of which carried an amplified MET gene, by Southern blot analysis and/or competitive polymerase chain reaction using eight DNA markers. Although the extent of the amplified domain varied, the close vicinity of the MET gene was the only region consistently amplified in these glioblastomas. A yeast artificial chromosome (YAC) contig of 900 kb was refined spanning the amplified region flanking the MET gene. The YAC inserts were subcloned into 59 cosmids, which were used for exon trapping. Eight sequences were identical to parts of the genes MET and CAPZA2 (human actin capping protein alpha-subunit). Two newly identified exons and the CAPZA2 exons were amplified in tumor TX3095, which retains an amplified MET gene. The new exons were localized close to MET and CAPZA2. Characterization of the clones, which were termed glioma-amplified sequence (GAS)7-1 and GAS7-2, showed an open reading frame and a different expression pattern in multiple human tissues. This study reports the identification of a cluster of amplified genes including two novel genes in a region amplified in more than 20% of glioblastomas. PMID- 9402968 TI - The human lysyl oxidase-like gene maps between STS markers D15S215 and GHLC.GCT7C09 on chromosome 15. AB - The lysyl oxidase-like (LOXL) gene is a new member of the lysyl oxidase family, a copper-dependent enzyme that is implicated in the crosslinking of collagen and elastin fibers. We have mapped the LOXL gene to chromosome 15q23, between STS markers D15S215 and GHLC.GCT7C09. This position corresponds to the q23 locus, not to the q24-25 locus suggested in a preliminary report. PMID- 9402969 TI - Pepsinogen polymorphism in the Indian population and its association with duodenal ulcer. AB - To date, there have been few studies on pepsinogen polymorphism. The present study examines the polymorphism of pepsinogen by PAGE in 155 duodenal ulcer cases and 92 control subjects. The Indian population presents a higher frequency of the B phenotype (associated with absence of the pg 5 fraction) and the C haplotype compared to other populations. Heterozygotes, in particular AC phenotypic individuals, are found to be associated significantly with the disease compared to control subjects. All the genes of the multigene complex controlling pepsinogen polymorphism seem to be interacting, thereby leading to such an association. Thus, studies at the gene level may be helpful in explaining the genetic etiology and heterogeneity of duodenal ulcer disease. PMID- 9402970 TI - Assignment of human plasma methylumbelliferyl-tetra-N-acetylchitotetraoside hydrolase or chitinase to chromosome 1q by a linkage study. AB - Plasma methylumbelliferyl tetra-N-acetylchitotetraoside hydrolase or chitinase (CHIT) might play a role in degrading the chitin wall of some microorganisms. In about 6% of Caucasian people the enzyme shows pseudodeficiency (defined as very low activity without apparent symptoms). We have mapped this locus by linkage analysis to the marker D1S306 (z = 4.00 at theta M = F = 0.0) on chromosome 1q between the flanking markers D1S191 and D1S245 in the area of 1q31-1qter. PMID- 9402971 TI - Genotype-phenotype correlation in cystic fibrosis patients compound heterozygous for the A455E mutation. AB - Cystic fibrosis (CF) has a high incidence in the French-Canadian population of Saguenay Lac-Saint-Jean (Quebec). The A455E mutation accounts for 8.3% of the CF chromosomes. This mutation was shown to be associated with a milder lung disease in the Dutch population. Twenty two CF patients distributed in 17 families and compound heterozygotes for the A455E mutation have been followed at the Clinique de Fibrose Kystique de Chicoutimi. Fourteen patients also carried the delta F508 mutation while the remaining eight patients had the 621 + 1G-->T mutation. Each patient was matched by sex and age to a patient homozygous for the delta F508 mutation. The pairs were analyzed for several clinical and laboratory variables. The A455E compound heterozygotes were diagnosed at a later age (P = 0.003) and had chloride concentrations at the sweat test lower than those homozygous for the delta F508 mutation (P = 0.007). More patients were pancreatic sufficient (P = 0.004). They had a higher Shwachman score (P = 0.001) and better pulmonary function tests (P < 0.02). CF patients compound heterozygous for the A455E mutation have a milder pancreatic and lung disease than the delta F508 homozygotes. Therefore, the A455E should be associated with a better prognosis. PMID- 9402972 TI - Genetic linkage study of a major susceptibility locus (D2S125) in a British population of non-insulin dependent diabetic sib-pairs using a simple non isotopic screening method. AB - A polymorphic microsatellite marker (D2S125) was recently reported to show significant linkage to non-insulin dependent diabetes mellitus (NIDDM) in a population of Mexican-American affected sib-pairs. We have used a simple non isotopic screening technique employing the polymerase chain reaction (PCR) with a biotinylated primer to study the genetic linkage and allele frequency distribution of the D2S125 marker in a population of 109 British NIDDMs (62 possible affected sib-pairs). The analysis provided no evidence for linkage of the D2S125 marker in the British subjects (MLS = 0.029, P > 0.05). The PCR screening method used proved to be a convenient and reliable alternative to the radiolabelling of PCR products. PMID- 9402973 TI - Sequence analysis of long FMR1 arrays in the Japanese population: insights into the generation of long (CGG)n tracts. AB - The human fragile-X syndrome is associated with expansions of a (CGG)n triplet repeat within the FMR1 gene. Whilst normal FMR1 arrays consist of variable numbers of (CGG)7-13 blocks punctuated with single AGG triplets, unstable arrays contain longer blocks of uninterrupted (CGG)n. The degree of instability, and subsequent risk of expansion to the fragile-X mutation, is dependent upon the length of this uninterrupted repeat. Detailed analyses of normal FMR1 array structures suggest that longer uninterrupted blocks of repeat could arise either through a process of gradual slippage or a more dramatic loss of an intervening AGG triplet. Up to 15% of Japanese and Chinese individuals have FMR1 triplet arrays centred on 36 repeats in length, a modal group not found in Caucasians. As longer FMR1 arrays have been associated with high-risk fragile-X haplotypes in some populations, we investigated the nature of these larger arrays. Sequence analysis revealed that the unusual length is due to the presence of a novel (CGG)6 block within the array. Several haplotypically related arrays contain blocks of (CGG)16 or (CGG)15, consistent with the fusion of adjacent (CGG)9 and (CGG)6 blocks after loss of the intervening AGG triplet. This is compatible with inferences from the Caucasian population that AGG loss is a mechanism by which long blocks of identical repeats are generated. PMID- 9402974 TI - Asynchronous replication of p53 and 21q22 loci in chronic lymphocytic leukemia. AB - In this study, in order to evaluate the replication pattern and the cell cycle dynamics of normal and malignant cells from patients with chronic lymphocytic leukemia, we applied the FISH technique with the p53 gene. Asynchrony was determined by the presence of one single and one set of double dots in the same cell. The rate of asynchronous replication was significantly higher in malignant cells than in normal cells (a mean of 28 vs 13, respectively, P = 0.023). There were proportionately more cells with two single dots among the normal cells (P = 0.0047). These results probably reflect the changes in gene replication and cell cycle progression that occur in malignant cells. PMID- 9402975 TI - Dual blastomere analysis improves reliability of preimplantation trembler mouse diagnosis. AB - Dual blastomere biopsy and independent blastomere analysis dramatically improved preimplantation diagnostic reliability as confirmed by testing the remaining biopsied eight-cell mouse embryo. The autosomal dominant trembler mouse point mutation was selected as a model for human preimplantation diagnosis because: (1) single cell assay failure is predicted to be the highest when testing autosomal dominant mutations; (2) point mutations represent the most common of all mutation categories and the most demanding mutation to assay reliably; and (3) the trembler mouse point mutation in peripheral myelin protein 22 (Pmp22) is a model of human Charcot-Marie-Tooth type 1A disease. Mathematical models predict our experimental results assuming amplification of 80% of each target allele as well as trembler sperm DNA contamination in 1 of 44 normal biopsied single blastomeres. Single blastomere analysis correctly predicted the genotype in only 84% of embryos that would have been implanted as normal. In contrast, when independent tests of both biopsied blastomeres agreed, test results were confirmed in 20 of 21 (95.2%) of the remaining six-cell biopsied embryos designated as normal. Thus, biopsied six-cell embryo confirmation demonstrated that dual biopsied blastomere analysis improved test reliability remarkably. PMID- 9402976 TI - Novel stop and frameshifting mutations in the autosomal dominant polycystic kidney disease 2 (PKD2) gene. AB - Autosomal dominant polycystic kidney disease (ADPKD) is one of the most frequent inherited disorders. The majority of cases are due to mutation of the PKD1 gene, on 16p13.3, while in most of the remainder the disease maps to the PKD2 locus, at chromosome 4q21-q23. Recently, the PKD2 gene has been positionally cloned and three nonsense mutations within the coding sequence of the gene identified. Here we report a systematic mutation screening of all 15 exons of the PKD2 gene in chromosome 4-linked ADPKD families, using heteroduplex and SSCP analyses. We have identified and characterized seven novel mutations, with a detection rate of approximately 90% in the population studied. All of the mutations result in the premature stop of translation: four nonsense changes and three deletions. The deletions are all frameshifting, of four T nucleotides in one case and one G nucleotide in the other two. All mutations are unique and are distributed throughout the gene without evidence of clustering. Comparison of specific mutations with the clinical profile in ADPKD2 families shows no clear correlation. PMID- 9402977 TI - Exclusion of PPEF as the gene causing X-linked juvenile retinoschisis. AB - X-linked juvenile retinoschisis (RS) is a progressive vitreoretinal degeneration localised in Xp22.1-p22.2. A human homologue of the retinal degeneration gene C (rdgC), a gene that in Drosophila melanogaster prevents light-induced retinal degeneration, was localised in the RS obligate gene region. We have tested the gene, designated PPEF in humans, as a candidate gene in RS patients using RT-PCR and the protein truncation test on RNA and SSCP on DNA. No mutations were identified, making it highly unlikely that PPEF is the gene implicated in RS. The data presented facilitate mutation analysis of the PPEF gene in other diseases which have been or will be localised to this region. PMID- 9402978 TI - Familial Mediterranean fever: clastogenic plasma factors correlated with increased O2(-)--production by neutrophils. AB - Familial Mediterranean fever (FMF) is an autosomal recessive disease predominantly affecting Armenians and non-Ashkenazi Jews. The disease begins in childhood with paroxysmal attacks of pain and fever accompanied by peritonitis, pleuritis, and synovitis. During the acute phase, there is a massive influx of polymorphonuclear leukocytes into the serosal membranes, connected with degranulation of the neutrophils and with secretion of lysosomal enzymes and pyrogenic substances. An increase in the lipoxygenase product, leukotriene B4, a chemotactic agent, and a decrease in the activity of the inhibitor of chemotaxis, C5a, in serosal fluids have been considered responsible. Previous work from our laboratories had shown that the chromosomal instability observed in blood cultures of patients with FMF is secondary to circulating clastogenic factors (CFs), and that the antioxidant enzyme superoxide dismutase, as well as lipoxygenase inhibitors, reduce the chromosome damaging effects. CFs are observed in chronic inflammatory diseases and in various other pathological conditions accompanied by oxidative stress. Similar clastogenic materials were found in supernatants of neutrophils and monocytes after a respiratory burst and were shown to contain lipid peroxidation products and cytokines. In the present study we compared the clastogenic effects exerted by plasma ultrafiltrates from 20 adult patients with FMF to the unstimulated O2- production of their neutrophils. In comparison to 20 age- and sex-matched controls, which were studied simultaneously, the O2- production by patient's neutrophils was routinely higher than that of controls. The clastogenic effects of patient's plasma, expressed as the number of chromosomal aberrations induced in test cultures of healthy donors, were correlated with the importance of O2- production by their neutrophils (r = 0.5235). Even if the relative contribution of disturbance in arachidonic acid metabolism, neutrophil activation, and CF formation in the disease process remains unclear, the demonstration of oxidative stress in this genetic disorder suggests the use of antioxidants and free radical scavengers, in particular during acute attacks, when the classical colchicine treatment is without effect. PMID- 9402979 TI - Serotonin transporter protein (SLC6A4) allele and haplotype frequencies and linkage disequilibria in African- and European-American and Japanese populations and in alcohol-dependent subjects. AB - The SLC6A4 locus encodes the serotonin transporter, which in turn mediates the synaptic inactivation of the neurotransmitter serotonin. Two PCR-formatted polymorphisms at this locus have been described, the first of which is a variable number tandem repeat located in exon 2, and the second a repeat sequence polymorphism located in the promoter region. The latter polymorphism alters transcriptional activity of SLC6A4, and has been reported to be associated with anxiety and depression-related traits. We studied allele frequencies, and computed haplotype frequencies and linkage disequilibrium measures, for these two polymorphisms in European-American, African-American, and Japanese populations, and in a set of alcohol-dependent European-American subjects. Allele frequencies for both systems showed variation, with significant differences overall for each system, and significant differences between each pair of populations for both systems. Linkage disequilibrium also varied among the populations. There were no significant differences in allele or haplotype frequencies between the European American population samples and alcohol-dependent subjects. The population differences demonstrate a potential for population stratification in association studies of either of these SLC6A4 polymorphisms. If genetic variation at this locus really is associated with behavioral variation, these results could reflect either different behavioral adaptations in different populations, or random genetic drift of a behaviorally important but selectively neutral polymorphism. PMID- 9402980 TI - No mitochondrial cytochrome oxidase (COX) gene mutations in 18 cases of COX deficiency. AB - Cytochrome c oxidase (COX) deficiency causes a variety of neuromuscular and non neuromuscular disorders in childhood and adulthood and can theoretically undergo either a nuclear or a mitochondrial (mt) mode of inheritance, making genetic counseling in COX deficiency particularly hazardous. In an attempt to determine the respective roles of mtDNA and nuclear DNA mutations in COX deficiency, we sequenced the three mitochondrially encoded COX subunits (COXI-III) in a series of 18 patients with isolated COX deficiency, especially as COXI-III code for the catalytic site of the enzyme. We failed to detect any deleterious mutations in this series. Moreover, no mtDNA deletion was observed and sequencing of the flanking tRNA genes involved in the maturation of the COX transcripts failed to detect deleterious mutations as well. The present study supports the view that the disease-causing mutations do not lie in the mt genome but, rather, in the nuclear genes encoding either the COX subunits or the proteins involved in assembly of the complex and suggests a recurrent risk of 25% rather than other modes of inheritance in COX deficiencies. PMID- 9402981 TI - Fragile site or dissociation of a t(Y;13)? PMID- 9402983 TI - A pharmacophore model of tautomycin, an inhibitor of protein phosphatases 1 and 2A. PMID- 9402984 TI - Amphistin, a new melanogenesis inhibitor, produced by an actinomycete. AB - A new melanogenesis inhibitor, named amphistin, was isolated from the fermentation broth of an actinomycete strain KP-3052. Amphistin was purified from the culture filtrate by the combination of cation exchange, gel filtration, and aminosilyl silica gel chromatographic methods. The structure of amphistin was elucidated as gamma-(beta-histidinoalanino)homoalanine by NMR experiments including 1H-15N HMBC experiment and other spectroscopic analyses. Amphistin inhibited the melanogenesis of B16 melanoma cells at concentration of 6.8 microM. PMID- 9402985 TI - BE-32030 A, B, C, D and E, new antitumor substances produced by Nocardia sp. A32030. AB - New antitumor substances, designated BE-32030A, B, C, D and E, were isolated from the culture broth of Nocardia sp. A32030. The active principles were extracted from the mycelium by methanol and purified by Sephadex LH-20 and reversed-phase column chromatographies and finally by reversed-phase HPLC. BE-32030A, B, C, D and E inhibited the growth of P388 murine leukemia, DLD-1 human colon cancer, PC 13 human lung cancer and MKN-45 human stomach cancer cell lines. PMID- 9402986 TI - Nocardicyclins A and B: new anthracycline antibiotics produced by Nocardia pseudobrasiliensis. AB - Nocardicyclins A (1) and B (2), new anthracycline antibiotics have been isolated from the mycelial cake of Nocardia pseudobrasiliensis IFM 0624 (JCM 9894). The molecular formulae of 1 and 2 have been determined as C30H35NO11 and C32H37NO12, respectively, and the structures were characterized by 1- and 8-methoxyl groups, a 10-carbonyl group and a novel carbon-methylated aminosugar constituent. Nocardicyclin A (1) exerts cytotoxic activity against L1210 and P388 leukemia. Nocardicyclins A (1) and B (2) are active against Gram-positive bacteria including Mycobacterium spp. and Nocardia spp., but inactive against Gram negative bacteria. PMID- 9402987 TI - A new antiherpetic agent, AH-1763 IIa, produced by Streptomyces cyaneus strain no. 1763. AB - A new antiherpetic agent, AH-1763 IIa, was isolated from the culture broth of strain No. 1763 identified as Streptomyces cyaneus. It was purified through column chromatographies of Diaion HP-10 and silica gel. The structure was determined to be 11-hydroxy-5-methyl-2-(2-hydroxy-1-methylpropyl)-4H-anthraceno [1,2-b]pyran-4,7,12-trione by several spectroscopic experiments, that is a new antibiotic belonging to pluramycin-group. PMID- 9402988 TI - CJ-12,954 and its congeners, new anti-Helicobacter pylori compounds produced by Phanerochaete velutina: fermentation, isolation, structural elucidation and biological activities. AB - Seven new phthalide compounds with anti-Helicobacter pylori activities were isolated from the basidiomycete Phanerochaete velutina CL6387. The two most potent phthalide compounds, CJ-12,954 and CJ-13,014, have MICs of 5 ng/ml. The structure-activity relationship shows that the presence of a spiroketal part in addition to the phthalide part, greatly enhances the activity. The phthalide compounds appear to be specific for H. pylori, since they did not show antibacterial activities when tested against a panel of other microorganisms. PMID- 9402989 TI - Novel anthelmintic metabolites from an Aspergillus species; the aspergillimides. AB - Two members of a novel class of anthelmintics, the aspergillimides, have been isolated from the Aspergillus strain IMI 337664. This novel fungus also produced two known and one structurally novel paraherquamide. This paper describes the fermentation, isolation, structure elucidation and anthelmintic activity of aspergillimide (VM55598, 1), 16-keto aspergillimide (SB202327, 2), and the paraherquamides VM54159 (3), SB203105 (4) and SB200437 (5). The aspergillimides are equivalent to paraherquamides which have lost both the dioxygenated 7 membered ring and the phenyl ring to which this is fused; gaining in their place a C8-keto group. SB203105 is the first example of a 4-substituted paraherquamide. PMID- 9402990 TI - Involvement of afsA in A-factor biosynthesis as a key enzyme. AB - The afsA gene of Streptomyces griseus has been postulated to encode a key enzyme for A-factor biosynthesis from primary metabolites commonly present in Streptomyces strains. Escherichia coli cells harboring afsA under the control of the T7 promoter specified distinct A-factor activity in the culture broth, as determined by induction of streptomycin production and aerial mycelium and spore formation in an A-factor-deficient S. griseus mutant strain. Production of the substance(s) having A-factor activity was inhibited by cerulenin, an inhibitor of fatty acid biosynthesis. These observations suggest that afsA encodes a key enzyme in the A-factor biosynthetic pathway in which a beta-keto acid derived from fatty acid biosynthesis and a glycerol derivative serve as precursors. PMID- 9402992 TI - New dihydro and tetrahydro derivatives of desmycosin. III. The opening of oxirane ring of 12,13-epoxydesmycosin. AB - Opening the oxirane ring of 12,13-epoxydesmycosin dimethylacetal (1) by catalytic hydrogenation gave the 10,11-dihydro-12,13-epoxy derivative (3) as the main product. Reductive oxirane cleavage was accomplished with dissolved metal (Zn) giving the 10,13-dihydro-13-hydroxy compound (6). Mild acid hydrolysis of 6 gave expected 10,13-dihydro-13-hydroxydesmycosin (8), but hydrolysis of 3, under the same conditions, gave three tautomeric desepoxy products. PMID- 9402991 TI - Inhibition of anchorage-independent growth of tumor cells by IT-62-B, a new anthracycline. AB - IT-62-B, a new anthracycline isolated from fermentation broths of Streptomyces sp. IT-62, reversed certain tumor cell phenotypes in vitro including some of human origin. The observed normal phenotypes were anchorage dependence of cell growth, flattened cell morphology and restoration of actin stress fibers. The extent of the anchorage dependence of cell growth induced by IT-62-B was generally greater than that by doxorubicin or pirarubicin. The cell-flattening effect of IT-62-B on cells of T24 (human bladder), but not on C-33A (human cervix), accompanied inhibition of fos gene expression. T24 cells, once flattened by IT-62-B, retained their flat morphology even in drug-free, fresh medium and eventually died in several days. IT-62-B, unlike doxorubicin, only slightly inhibited the topoisomerase II reaction in vitro and DNA synthesis in isolated cell nuclei. PMID- 9402993 TI - Chemical synthesis and structural study of lincomycin sulfoxides and a sulfone. AB - Oxidation of lincomycin with dimethyldioxirane resulted in the sulfoxide glycosides 3a and 3b, whose treatment with osmium tetraoxide and N methylmorpholine-N-oxide afforded the same sulfone; 4. According to FAB-MS and CD investigations, the absolute configuration of the sulfur atom in 3a and 3b is R and S, respectively. The new, unsaturated antibiotic analog (6) derived from clindamycin exists in the 4C1 conformation. The antibiotic activities of the synthesized compounds were also studied. PMID- 9402994 TI - delta-Indomycinone: a new member of pluramycin class of antibiotics isolated from marine Streptomyces sp. PMID- 9402995 TI - Macrosphelides C and D, novel inhibitors of cell adhesion. PMID- 9402996 TI - Discovery of methyl-2-(2'-hydroxyphenyl)-2-oxazoline-4-carboxylate as a secondary metabolite from Actinomadura sp. PMID- 9402997 TI - Corynecandin: a novel antifungal glycolipid from Coryneum modonium. PMID- 9402998 TI - The identification of 1,6'- and 1,3"-di-N-(L-4-amino-2-hydroxybutyryl) derivatives of kanamycin as synthetic byproducts of amikacin. PMID- 9402999 TI - Phomopsidin, a new inhibitor of microtubule assembly produced by Phomopsis sp. isolated from coral reef in Pohnpei. PMID- 9403000 TI - Induction of mixed allogeneic chimerism for leukemia. AB - Hybrid (C56BL/6 x DBA) (BDF1; H-2b/H-2d) mice bearing the P815 leukemia (H-2d) were grafted with a (CBA x C57BL/6)F1 (CBF1; H-2k/H-2b) cell suspension, comprising bone marrow cells (BMC; 25 x 10(6)/mouse) and spleen cells (SC; 55 x 10(6)/mouse) on day-4, then treated with cyclophosphamide (200 mg/kg) on day-2 and finally grafted once more with CBF1 cells (25 x 10(6) BMC + 7 x 10(6) SC) on day 0. Allogeneic cell graftings performed in this way induced durable mixed hematopoietic chimerism and significantly prolonged the survival of recipients, compared with that of leukemia-bearers grafted with syngeneic cells. The results obtained raise the possibility of using allogeneic hematopoietic tissue transplantation in combination with non-lethal cytoreductive therapy to induce a long-lasting graft-vs-leukemia effect. PMID- 9403002 TI - Allelic loss of the FMS gene in acute myeloid leukaemia. AB - The FMS proto-oncogene encodes for the colony stimulating factor-1 receptor expressed on monocytes and B lymphocytes within the peripheral blood system. Allelic loss of the FMS gene occurs in patients with refractory anaemia and the 5q- syndrome associated with the myelodysplastic syndromes. To determine the frequency of FMS gene loss in patients with myeloid malignancy, 50 DNA samples from patients with acute myeloid leukaemia (AML) and 30 samples from haematologically normal samples were analysed using a quantitative Southern blotting technique. Allelic loss of one allele (hemizygous) was detected in five of 18 samples of AM-M4 and eight of 27 samples of AML M1, M2 and M3. In addition, loss of both FMS alleles (homozygous) was demonstrated in three of 18 samples of AML M4 and 0127 samples of AML M1, M2 and M3. One patient with AML M5 and one with AML M6 were assessed although no allelic loss of FMS was detected. Three samples from patients with secondary AML were also analysed and hemizygous loss was detected in one case. Homozygous or hemizygous loss of FMS was not detected in any of 30 DNA samples isolated from haematologically normal individuals. These data indicate that loss of the FMS gene is common in AML, with an increased frequency in those patients with AML subtype M4. PMID- 9403001 TI - The FEL (AF-4) protein donates transcriptional activation sequences to Hrx-Fel fusion proteins in leukemias containing T(4;11)(Q21;Q23) chromosomal translocations. AB - The t(4;11) chromosomal translocation marks a subset of acute lymphoblastic and secondary myeloid leukemias. It results in the fusion of the FEL (AF-4) gene on chromosome band 4q21 with the HRX (MLL) gene on chromosome band 11q23. This translocation results in the expression of fusion transcripts from both translocated chromosomes, with the derivative 11 product (fusing the amino terminal third of the Hrx protein to the C-terminal two-thirds of the Fel protein) thought to be involved in leukemic transformation. The mechanism of transformation by Hrx-Fel in leukemic cells, however, is unknown and the specific leukemogenic contributions of Fel have not been defined. In this study, we demonstrate that Fel is capable of activating transcription from a minimal adenoviral E1b promoter as a Gal4-Fel fusion protein in transient transcriptional assays. The Fel transactivating sequences were localized to amino acids 365-572 which are consistently retained by Hrx-Fel fusion proteins created by t(4;11) translocations in leukemias. Furthermore, we demonstrate that the transactivation properties of Fel vary in different cell types. While Gal4-Fel constructs strongly activated transcription in Cos-7 cells and the MCF-7 breast tumor cell line, they displayed low to no activity in the precursor B-cell line REH, breast tumor cell line Gl-101A and epithelial-derived A431 cells. These data are consistent with a potential role of Hrx-Fel as a chimeric transcription factor in which Fel contributes transcriptional effector properties and suggest the requirement for cell-specific accessory factors. PMID- 9403003 TI - Growth inhibition of B-cell precursor acute lymphoblastic leukemia cell lines by monocytes: a role for prostaglandin E2. AB - Leukemic cell lines have proven invaluable in the molecular analysis of recurring chromosomal translocations but the optimal methods for leukemia cell line establishment are unknown. During in vitro culture, most B-cell precursor acute lymphoblastic leukemia (BCP-ALL) cells die within 1 week at least partially mediated by inhibitors elaborated by peripheral blood mononuclear cells (PB MNCs) present within the leukemia sample. In experiments reported here, cyclooxygenase inhibitors (indomethacin and meclofenamic acid) blocked the PB MNC-mediated inhibition of BCP-ALL proliferation. Also, prostaglandin E2 (PGE2) was detected in supernatants from PB MNC cultures. When PGE2 was mixed directly with BCP-ALL cells, proliferation decreased significantly. Under the culture conditions used, PB MNCs secreted PGE2 which appears to be one of the major inhibitors of BCP-ALL growth in vitro. PMID- 9403004 TI - A novel method for direct and fluorescence independent determination of drug efflux out of leukemic blast cells. AB - Multi drug resistance (MDR) is often due to an increased efflux of anti cancer drugs out of leukemic blast cells. Efflux assays are used to get an impression of functional resistance in those cells. Dyes like rhodamine 123 or 3'3' diethyloxocarbocyanine iodide are commonly used for this purpose. A major known disadvantage is that dyes do not behave like cytotoxic drugs in efflux experiments. Assays using the self fluorescence of drugs like anthracyclines can not reveal a real impression of intracellular or effluxed drug due to quenching of the drug fluorescence in the nuclei of the cells. We have developed a reproducible and sensitive assay for direct and quantitative determination of drug efflux out of blast cells. This was done by a novel double radioactive labelling using a 3H-labelled drug and 14C-labelled sucrose as extracellular marker. So this assay can be applied to every drug of interest. Quenching of fluorescence is also by-passed with this technique as well as protracting washing or silicon oil procedures. As a model system we used the T-lymphoblastoid cell line CCRF CEM and its resistant sublines vincristine 100 and adriamycin 5000. The results were also transferable to clinical specimens of leukemic patients. In conclusion our assay may be used for precise and direct efflux measurement of a broad range of anti-cancer drugs in clinical MDR evaluation. PMID- 9403005 TI - Characterization and sensitivity to interleukin 2 and interferon alpha of leukemic cells from a patient with large granular lymphocytic leukemia associated with chronic active Epstein-Barr virus infection. AB - A patient presented with chronic large granular lymphocyte leukemia associated with chronic active Epstein-Barr virus infection (CAEBV). Cell cycle analysis revealed a minimal growth compatible with chronic lymphocytic leukemia After 5 months of treatment, the patient died from acute transformation of the leukemia. Cell harvested during chronic phase were analyzed for sensitivity to interleukin 2 (IL-2) and interferon alpha (IFN alpha) in vitro by means of surface phenotyping and cell cycle assay. IL-2 induced remarkable growth of the cells, whereas IFN alpha did not confer a growth advantage. Since IFN alpha was expected to have no growth induction effect on the leukemia cells, it was administered to the patient to treat the CAEBV. PMID- 9403006 TI - Retroviral transfer of herpes simplex virus-thymidine kinase and beta galactosidase genes into U937 cells with bicistronic vector. AB - In this study we describe a new retroviral vector utilizing an internal ribosome entry site (IRES) from encephalomyocarditis virus to co-express two genes. One is the herpes simplex virus type 1 thymidine kinase gene (HSV-TK) which induces sensitivity to ganciclovir, and the second is the bacterial beta-galactosidase gene (LacZ) which was revealed by an histochemical staining with 5-bromo-4-chloro 3-indolyl-beta-D-galactopyranoside (X-Gal). We engineered the U937 human cell line to co-express both genes and monitored transduced cells using X-Gal staining. Several transduced clones were selected. The clones exhibiting X-Gal positive cells were sensitive to ganciclovir treatment (1 microgram/ml) while X Gal negative clones were not. Monoclonal cell lines showed a single copy of the provirus integrated in their genome with the TK-IRES-LacZ sequence stably inserted in all clones. The band distribution pattern of the proviral DNA differed only at the long terminal repeat (LTR) level. Northern blot analysis of an X-Gal positive/ganciclovir sensitive clone showed an mRNA band of 6 kb with both LacZ and TK probes. An X-Gal negative/ganciclovir resistant clone was negative with both probes. This report shows: (1) a therapeutic gene can be linked to a marker gene by an IRES element achieving equivalent expression of both proteins; (2) the co-expression of a marker gene makes fluorescein-di-beta-D galactopyranoside staining possible, and consequently separation of cells expressing the LacZ gene by fluorescence activated cell sorting. Thus the cells expressing the HSV-Tk gene are enriched; (3) the use of a marker gene such as LacZ could open up interesting perspectives in gene therapy protocols because of the opportunity to monitor the transduced cells using a simple cytochemical stain. PMID- 9403007 TI - Camptothecin causes cell cycle perturbations within T-lymphoblastoid cells followed by dose dependent induction of apoptosis. AB - We have investigated the effect of the anticancer compound, camptothecin on Jurkat T-cells, a lymphoblastoid leukemic cell-line. Exposure to low concentrations led to rapid cessation of DNA (more than 95%) and RNA (more than 75%) synthesis. Perturbations to the cell cycle were observed following exposure which caused a significant accumulation of cells within G1 (P = 0.03) with a concomitant decrease in G2/M (P = 0.025). Concentrations below 0.1 microM could inhibit DNA synthesis but not induce apoptosis. Induction of apoptosis was dose dependent and could be detected as early as 3 h post exposure. The apoptotic population appeared to be derived from G1 and S-phase cells but not G2/M, coinciding with the cell cycle compartments in which DNA and RNA polymerases function. However, direct inhibition of DNA polymerase alone was not shown to be associated the induction of apoptosis or with a decrease in susceptibility to camptothecin-induced cell death. The effects of camptothecin on Jurkat T-cells and the potential mechanisms involved are discussed in the context of observations made in other transformed cell lines. PMID- 9403008 TI - Mitochondrial ultracondensation, but not swelling, is involved in TNF alpha induced apoptosis in human T-lymphoblastic leukaemic cells. AB - Mitochondrial permeability transition (PT) pore opening and mitochondrial swelling have been reported in association with apoptosis. Conformational alterations of mitochondria induced by tumour necrosis factor-alpha (TNF alpha), and the association with TNF alpha-induced apoptosis, were, therefore, studied in the human acute T-lymphoblastic leukaemia (T-ALL) cell line, CCRF-CEM and its vinblastine-resistant CEM/VLB100 cell line by transmission electron microscopy (TEM). The CEM/VLB100 cell line possessed more condensed (C phase) mitochondria in the resting state compared with its parental cell line, consistent with increased activity of the mitochondrial electron transport chain (ETC). Following exposure to TNF alpha, conformational alterations of mitochondria occurred in both apoptotic and non-apoptotic cells. Orthodox (O phase) mitochondria in non apoptotic cells underwent C-phase, transitional O-phase and slightly swollen (S phase) conformational changes. TNF alpha-induced mitochondrial swelling was a late event and was found to a far lesser extent than mitochondrial condensation. No swollen mitochondria were observed in apoptotic cells. Ultracondensed (UC phase) mitochondria were observed in cells undergoing both TNF alpha-induced and spontaneous apoptosis and were seen when TNF alpha-induced apoptosis was inhibited by 3-methyladenine (3MA). The structural integrity of UC phase mitochondria persisted through the apoptotic process. We conclude that TNF alpha induced mitochondrial swelling and apoptosis are separate events. Mitochondrial ultracondensation is associated with the processes signalling apoptosis and is not a result of TNF alpha-induced apoptotic shrinkage. PMID- 9403010 TI - Getting plant toxins to fuse. PMID- 9403011 TI - HL-60 growth suppression by citrate-seric growth factor interaction. PMID- 9403009 TI - The prognostic significance of chromosomal analysis and immunophenotyping in 117 patients with de novo acute myeloid leukemia. AB - Chromosomal abnormalities is one of the most important prognostic factors in acute myeloid leukemia (AML). Other parameters which may influence the prognosis include age, French-American-British-type, clinical variables and possibly the expression of certain immunophenotypic surface makers. However, only rarely has the expression of these markers been analyzed in multivariate models including the information from cytogenetics and clinical variables. We conducted a retrospective study of 117 consecutive adult patients with de novo AML diagnosed and treated in our institution during a 6-year period. Following standard induction chemotherapy with daunomycin and cytosine arabinoside 75 patients (64%) achieved complete remission (CR). The overall 5 year survival rate was 23% and, for patients achieving CR, 30%. When all patients were analyzed age, chromosomal aberration and lack of CD33 expression were of independent prognostic value. The overall 5 year survival rate was 28% for patients aged 55 years or younger, 25% for patients aged 56-65 years and 4% for those > 65 years, P = 0.041. Patients with good-risk chromosomal abnormalities presented an overall 5 year survival of 36%, compared to 25% in patients with normal karyotype, 22% in patients with intermediate risk abnormalities and 5% in patients with poor-risk abnormalities, P = 0.004. Patients with CD33+ myeloblasts had an overall survival of 25% at 5 years compared to 0% in the CD33- patients, P = 0.021. Analysis of the expression of CD7, CD34 and terminal deoxynucleotidyl transferase on myeloblasts had no impact on overall survival in a multivariate analysis. Thus, this study confirmed the prognostic value of age and cytogenetic risk group and defined CD33 as a novel factor of independent prognostic importance in adult de novo AML. PMID- 9403012 TI - An atypical case of plasma cell leukemia. PMID- 9403013 TI - Activity of high-dose cyclophosphamide in the treatment of childhood malignant gliomas. AB - Seventeen patients less than or equal to 20 years of age with newly diagnosed (n = 10) or recurrent (n = 7) malignant gliomas (anaplastic astrocytoma and glioblastoma multiforme) were treated with cyclophosphamide in association with hematopoietic cytokines (GM-CSF or G-CSF). Cyclophosphamide was given at a dose of 2 g/m2 daily for 2 days at 4-week intervals. Toxicity consisted of grade IV neutropenia and thrombocytopenia in 95% and 48% of cycles, respectively. There were no cyclophosphamide-related cardiac, pulmonary, or urothelial toxicities observed. Four of 10 patients with newly diagnosed disease demonstrated responses (three complete and one partial responses; one CR was only of 2 months duration). None of the seven patients with recurrent tumors demonstrated a response. We conclude that high-dose cyclophosphamide warrants further evaluation in children with newly diagnosed malignant glioma. PMID- 9403014 TI - Subcutaneous sacrococcygeal myxopapillary ependymoma. AB - We report an 8-year-old boy with a primary subcutaneous sacrococcygeal ependymoma, a rare tumor that is thought to arise in embryologic rests. The lesion was completely removed in our patient, who has been followed without recurrence for 20 months. Our experience, together with that of the other 15 cases in the world literature, supports surgical excision as the mainstay of treatment. PMID- 9403015 TI - Electronic measurement of compliance with mercaptopurine in pediatric patients with acute lymphoblastic leukemia. AB - Twenty-four pediatric patients with acute lymphoblastic leukemia (ALL) on maintenance therapy were evaluated for their compliance with taking their prescribed doses of oral mercaptopurine (6-MP). PROCEDURE AND RESULTS: We utilized the Medication Event Monitoring System (MEMS; Aprex Corporation, Fremont, CA) for the study. Compliance was defined as the number of days doses were taken as a percentage of the total number of days doses were prescribed during the study period. The mean age of the patients was 7.3 years (range 2.6 17.2 (years). Patients were evaluated for a mean of 44 days (range 15-94 days). Thirty-three percent of patients (8) took less than 90% and 17% (4) took less than 80% of their prescribed pills. Eight patients were also evaluated for a difference in compliance between morning and evening administration. For the comparison of compliance between a morning vs. an evening schedule a tren toward improved compliance in the evening was found. Five patients had an increase and one patient a decrease in compliance with an evening schedule (differences ranged from 0.2% to 51.3%), with two patients having 100% compliance on both schedules. CONCLUSIONS: Our data raise concern that a significant proportion of pediatric patients are non-compliant with pill taking and demonstrate that the timing of administration of 6-MP in children with ALL may be crucial in some patients and supports the hypothesis that evening administration of 6-MP is associated with a lower risk of relapse. PMID- 9403016 TI - Cytogenetic evidence for a less malignant leukemic cell population in the central nervous system in a critical case of acute myeloblastic leukemia. AB - BACKGROUND: With the exception of a single study the cytogenetic aspects of leukemic cells in the central nervous system (CNS) have not been investigated. PATIENTS AND RESULTS: During the course of a work-in-progress on the chromosomal constitution both of the spinal fluid and of bone marrow (BM) in children with acute myeloblastic leukemia (AML), we have observed a unique case of AML and CNS leukemia (CNSL) at diagnosis. The patient showed the simultaneous presence at diagnosis of a 46 cytogenetic line in the spinal fluid and a 47 (+8) cell line in the BM, present in the great majority of the metaphases examined. DISCUSSION: This observation allows hypotheses on the relationship between BM and CNS disease in AML. Regardless of the pathogenetic mechanism, the cytogenetic findings of the present case clearly suggest that the leukemic population in the CNS compartment represents a less malignant cell process compared to the BM leukemic population. This easily fits in with the usually less malignant course of CNSL in AML. CONCLUSION: The foregoing findings may have critical pathogenetic and therapeutic implications. PMID- 9403017 TI - Home intravenous antibiotic treatment for febrile episodes in immune-compromised pediatric patients. AB - The purpose of this work was to assess the feasibility of home intravenous antibiotic treatment (HIAT) for febrile episodes in immune-compromised (neutropenic, splenectomized), low-risk pediatric patients. Thirty hematology oncology patients who presented to our emergency room from January 1993 to January 1995 and who suffered from a febrile episode and were considered at low risk for septic complications were immediately discharged on HIAT. Patients were followed for at least 3 weeks after recovery. Patients and parents were retrospectively questioned about adverse effects and about their degree of satisfaction with home treatment. Patients who required hospitalization during this period were considered unresponsive to HIAT and were analyzed for causes and adverse effects. Thirteen out of 60 (22%) febrile episodes, or eight out of 42 (19%) episodes of fever and neutropenia eventually led to hospitalization. Pseudomonas species infections were associated with the highest rate of unresponsiveness (88%). A central venous catheter infection developed in two cases following HIAT (two cases out of 640 days of therapy). No other complications were identified. No infection-related morbidity was observed. Patients and parents were highly satisfied with HIAT and wanted to use it again, if necessary. Immediate discharge on HIAT for low-risk pediatric immune compromised patients suffering from a febrile episode is feasible, safe, and well accepted by patients and families. Patients who are found to have Pseudomonas infections should probably be hospitalized. Our results are preliminary and must be confirmed by a prospective, randomized trial before definite recommendations can be made. PMID- 9403018 TI - Emergent/urgent therapeutic irradiation in pediatric oncology: patterns of presentation, treatment, and outcome. AB - PURPOSE: We reviewed all pediatric cases referred for emergent/urgent therapy (requiring treatment within 48 hours) to identify frequency, patterns of presentation, and efficacy of therapy. We defined five categories of emergent/urgent therapy based on irradiated site and/or signs: Group I, spinal cord compression; Group II, respiratory compromise; Group III, infradiaphagmatic distress; Group IV, intracranial signs; Group V, pain. MATERIALS AND METHODS: From 2/1/88-3/1/ 94, 104 children with 115 problems were referred by specialists at the Children's Hospital of Philadelphia. Diagnosis, nature of the emergency, and response were examined. Responses were categorized as complete resolution, improvement or stabilization, and progression. RESULTS: The 104 children represented 12% of referrals during the study period. The most common tumors were CNS PNET and gliomas (20%); and neuroblastoma (20%). Forty-five problems occurred with newly diagnosed tumors and 70 after progression. Ninety-one episodes were managed with radiation therapy and 24 with other modalities. Patients with spinal cord/cauda equina (n = 33) compression improved (55%) or stabilized (30%). Patients with respiratory compromise from thoracic (n = 14) or abdominal (n = 5) disease had a response rate of 72%. Eight patients in group III had a 66% response. In Group IV (n = 16), 63% had complete responses and 19% had stabilization. Group V (n = 15) patients had a complete or partial response of 93%. CONCLUSION: Approximately 10% of children referred for radiation therapy required emergent/urgent treatment. Eighty percent of patients achieved stabilization or showed improvement in signs and symptoms, indicating that radiotherapy is a valuable and reliable component of multimodal care in such situations. PMID- 9403019 TI - Successful management with octreotide of a child with L-asparaginase induced hemorrhagic pancreatitis. AB - BACKGROUND: Octreotide is a synthetic somatostatin analogue which has been suggested for use in the management of acute pancreatitis. While studies have looked at octreotide use in the setting of pancreatitis due to chronic alcohol use or trauma, little is known of its role in management of drug induced acute pancreatitis; particularly in the pediatric setting. PATIENTS AND METHODS: We present a case of a 5 1/2-year-old white female who developed severe, necrotizing, hemorrhagic pancreatitis with pseudocyst formation secondary to L asparaginase use as a part of her therapy for acute lymphoblastic leukemia (ALL). She was managed initially with intravenous fluids, bowel rest, nasogastric suctioning, parenteral narcotices, and broad spectrum antibiotics. In addition, within 12 hours of admission to The Children's Hospital (TCH) in Denver, Colorado, she began therapy with octreotide (5 micrograms/kg/day IV divided b.i.d.). With this management, her pseudocyst decompressed without need for surgical intervention, her pancreatitis fully resolved, and she recovered full pancreatic function without any long-term sequelae. CONCLUSION: Use of octreotide may have served a role in limiting the severity of the disease process in this case. Further studies need to be done to verify its usefulness in this setting. PMID- 9403020 TI - Recombinant human erythropoietin for the treatment of anemia in children with solid malignant tumors. AB - BACKGROUND: Cancer is often associated with chronic anemia which frequently requires blood transfusions. This study was performed to assess the efficacy and safety of r-HuEPO therapy in children with cancer. PATIENTS AND METHODS: Twenty five patients under 18 years of age with solid malignant tumors were treated with 150 U/kg/day of r-HuEPO 5 times weekly for 12 weeks. Response was defined as an increase of the baseline hemoglobin level by at least 2 g/dl. r-HuEPO patients were compared to 25 matched historical controls. RESULTS: Response was achieved in 72% of r-HuEPO patients. Hemoglobin level increased from 9.8 +/- 0.6 g/dl at baseline to 12.4 +/- 1.7 g/dl at the end of treatment in the r-HuEPO group and increased from 9.5 +/- 0.7 g/dl to 9.6 +/- 1.4 g/dl in the control group (P < .001, Student's t-test). Only 16% of patients receiving r-HuEPO required blood transfusions vs 96% of control patients (P < .001, Student's t-test), with mean units of blood transfused per patient being 0.35 in the r-HuEPO group and 3.56 in controls (P < .001, Student's t-test). There was a statistically significance improvement in Karnofsky's index in r-HuEPO patients. No adverse reaction related to r-HuEPO therapy was observed. CONCLUSIONS: r-HuEPO is a safe and effective means of increasing hemoglobin level and reducing blood requirements in children with solid malignant tumors receiving chemotherapy. PMID- 9403021 TI - Use of pamidronate in the management of acute cancer-related hypercalcemia in children. AB - PURPOSE: To determine whether pamidronate is a safe and effective agent for the treatment of severe hypercalcemia of malignancy in children. MATERIALS AND METHODS: A retrospective review of the charts of five children treated with pamidronate 1-2 mg/kg for severe, refractory hypercalcemia of malignancy. All children failed conventional therapy. Statistical analysis was done utilizing the two-tailed Student's t-test. RESULTS: All five children had complete resolution of their hypercalcemia in a predictable pattern within 24-48 hours. The average decrease in serum calcium was 1.63 mmol/L (6.54 mg/dl). (P < .01) The adverse effects were mild and transient, and consisted of hypocalcemia, hypophosphatemia, and hypomagnesemia. CONCLUSIONS: Pamidronate at a dose of 1 mg/kg is a safe and effective treatment for severe, refractory hypercalcemia of malignancy in children. PMID- 9403022 TI - Misleading leads: focal xanthogranulomatous pyelonephritis in childhood. AB - Xanthogranulomatous pyelonephritis is a morphologic variant of pyelonephritis. Focal disease is very rare and can be misdiagnosed. PMID- 9403023 TI - "Arteriovenous fistula: a complication following renal biopsy of suspected bilateral Wilms tumor". PMID- 9403024 TI - "Pancreatoblastoma in childhood: the role of alpha-fetoprotein". PMID- 9403025 TI - Interindividual variation in cytogenetic response to X-ray and colchicine measured with the cytokinesis-block micronucleus assay. AB - Interindividual variation in cytogenetic response to two different types of micronucleus (MN) inducer, X-rays (a clastogen) and colchicine (a spindle poison), was investigated in the peripheral blood lymphocytes of normal healthy donors by the cytokinesis-block MN method. The data for 124 donors between the ages of 19 and 80 years showed that the histogram of individual frequency of X ray (2 Gy)-induced micronucleated cells followed the normal distribution (Shapiro Wilks W-test) with a significant interindividual variance (ANOVA, p < 0.001). This was, however, not the case for colchicine (0.03 microgram/ml)-induced micronucleated cells. Instead, a skewed distribution illustrating interindividual variation was evident (ANOVA, p < 0.001). Statistical analysis of the effect of age and sex on MN incidence by using the Kruskal-Wallis test indicated that age affected the baseline and colchicine-induced MN incidences strongly but not the X ray-induced MN incidence. There was no effect of sex on the incidence of micronuclei induced by either agent. In order to avoid any possible effect of age on the MN index, data for young subjects aged less than 30 years old were analyzed separately. The results of this analysis again showed significant interindividual variations in baseline, X-ray-induced, and colchicine-induced micronucleated cell rates. Results of the correlation-coefficient analysis showed that neither X-ray-induced MN incidence nor colchicine-induced MN incidence was related to baseline MN incidence. No correlation between X-ray-induced and colchicine-induced MN incidences was also found by this analysis. These results suggest that interindividual variance in chromosomal response to mutagens in normal populations may be a real phenomenon, as is interindividual variance in baseline MN frequency, and that individual susceptibilities to the two different types of micronucleus inducers (X-ray and colchicine) are unrelated, and the baseline MN level is not of predictive value for the susceptibilities. PMID- 9403026 TI - New tester strains of Salmonella typhimurium lacking O6-methylguanine DNA methyltransferases and highly sensitive to mutagenic alkylating agents. AB - Salmonella typhimurium YG7104 and YG7108 are derivatives of the Ames tester strain TA1535, and have chromosomal deletions of the ogtST gene or both the ogtST and adaST genes, respectively. The ogtST and adaST genes encode O6-methylguanine DNA methyltransferases that are involved in the repair of DNA damage caused by alkylating agents. The sensitivities of these strains to 15 mutagens with different structures were tested and compared with those of the parent strain TA1535. Deletion of ogtST or ogtST plus adaST substantially increased the sensitivity of strain TA1535 to the mutagenicity of alkylating agents, such as N ethyl-N'-nitro-N-nitrosoguanidine, ethyl methanesulfonate or dimethylnitrosamine (DMN). Preincubation of the chemical with S9 mix and bacteria for 20 min at 37 degrees C before pouring them together on agar plates was not necessary to detect the mutagenicity of DMN when strain YG7104 or YG7108 was used as a tester strain. Introduction of plasmid pKM101 did not enhance but rather decreased the sensitivity of YG7104 and YG7108 to alkylating agents. Since the new strains are highly sensitive only to alkylating agents, they will be useful to detect the mutagenicity with high efficiency and to study the mechanism of mutagenesis induced by environmental alkylating agents. PMID- 9403027 TI - Mutations at codon 249 of p53 gene in human hepatocellular carcinomas from Tongan, China. AB - Codon 249 (exon 7) of the putative tumor suppressor gene p53 is a mutational hot spot for hepatocellular carcinoma (HCC) but not other tumors. DNA samples from primary HCC patients from Tongan, an area of high HCC incidence in China (> 40 per 100,000 population), were analyzed for specific mutations in codon 249 of the p53 gene using polymerase chain reaction (PCR)/restriction-digest methods and direct DNA sequencing. Seven of the 21 samples screened were found to have a point mutation at the third base position of codon 249 (AGG to AGT). The result is consistent with previous reports that the G-->T transversion is positively associated with the level of dietary aflatoxin B1 (AFB1) contamination, which has been implicated as one of the risk factors in Tongan area. Of the 7 HCC patients that contained the codon 249 point mutation, one was hepatitis B virus (HBV) negative. This is only the second documentation of an HCC patient harboring the p53 codon 249 mutation, who was HBV-negative. PMID- 9403028 TI - Comparison of alkaline single cell gel (Comet) and peripheral blood micronucleus assays in detecting DNA damage caused by direct and indirect acting mutagens. AB - The alkaline single cell gel (SCG) or 'comet' and peripheral blood micronucleus (pbMN) assay have been used to compare the effects of the direct acting mutagens, methyl methanesulfonate (MMS) and N-nitroso-N-methylurea (NMU), and the indirect acting mutagens, benzo[a]pyrene (BAP), cyclophosphamide (CP) 9, 10-dimethyl-1,2 benzanthracene (DMBA), and mitomycin C (MMC) in an inbred house mouse (Mus domesticus) strain. The alkaline SCG assay was able to detect DNA damage from direct acting mutagens. However, it appears that, even at the highest concentrations tested, the SCG assay was not able to detect DNA damage caused by 3 of 4 indirect acting mutagens tested. The exception was BAP. The pbMN assay was sensitive to DNA damage caused by both groups of mutagens. Multiple injections did not increase the sensitivity of the SCG assay to the indirect acting mutagen CP. Further, simultaneous injections of CP and MMS, in one experiment, resulted in significantly lower (p < 0.05) average DNA ratios and micronucleated polychromatic erythrocyte counts than those obtained after treatment with MMS alone. Although the SCG assay has been shown to be sufficiently sensitive to detect DNA damage caused by both direct and indirect acting mutagens in deermice (Peromyscus maniculatus) and bullheads (Ameiurus nebulosus), similar results are not seen in the inbred house mouse strain tested. PMID- 9403029 TI - Nitroreductase independent mutagenicity of 1-halogenated-2,4-dinitrobenzenes. AB - The 1-halogen substituted 2,4-dinitrobenzenes have been found to be mutagenic in Salmonella TA98 with an activity order of 1-fluoro > 1-chloro > 1-bromo > 1-iodo. This specific activity was not lowered in the nitroreductase deficient strain TA98NR and O-acetyltransferase deficient strain TA98/1,8-DNP6, indicating that the mutagenicity of these compounds is not dependent upon -NO2 reduction to -NHOH and its subsequent esterification. The activity was, rather, higher in the former and remained almost equal in the latter strain compared to TA98, suggesting these compounds to react directly with bacterial DNA. Further, the reaction of these halodinitrobenzenes with methionine, at physiological pH, resulting in the formation of 1-S-methyldinitrobenzene showed that these compounds have the ability to attack DNA directly through nucleophilic substitution of the halogen atom and the role of the bacterial nitroreductases for production of mutagenic lesions was not considered. It was, further, proposed that these compounds interact with DNA through SNAr mechanism instead of SN2 cited in literature. PMID- 9403030 TI - Mutations induced by monofunctional and bifunctional phosphoramide mustards in supF tRNA gene. AB - The relative mutagenicity, nature of the mutations and the sequence specificity of mutations induced by the bifunctional alkylating agent, phosphoramide mustard (PM) and a monofunctional derivative, dechloroethyl phosphoramide mustard (dePM), were analyzed by the Ames test and by an in vitro shuttle vector mutagenesis assay. Both PM and dePM increased the mutation frequency above background in either assay. However, on an equimolar basis, dePM was less mutagenic than PM. In the in vitro shuttle vector mutagenesis assay, sequencing demonstrated that about 40% of the mutant plasmids contained more than one mutation in the supF tRNA gene segment of the plasmid. About 70% of the mutations observed in dePM-treated plasmids were single base substitutions with A:T and G:C base pairs being mutated at equivalent rates. In contrast, only about 50% of the mutations observed in PM treated plasmids were single base substitutions, 80% of which involved G:C base pairs. Single base deletions and insertions were found in approximately equal proportions with both compounds; however, these lesions were in greater abundance in PM-treated plasmids. Putative hot-spots for mutation in the supF tRNA gene included base pairs at position 102 and 110 for PM and positions 170 and 171 for dePM. PMID- 9403031 TI - In vitro fertilization rate of mouse oocytes with spermatozoa from the F1 offspring of males irradiated with 1.0 Gy 137Cs gamma-rays. AB - Previous studies suggest that the spermatozoa from acutely irradiated male mice exhibit a reduced fertilization rate in vitro with the maximum decrease occurring for spermatozoa produced 6 weeks after irradiation (Y. Matsuda et al., Mutation Res. 142 (1985) 59-63). We have found that spermatozoa from unirradiated F1 males conceived 6 weeks after paternal F0 irradiation also exhibit a significantly reduced fertilization rate in vitro. After acute 137Cs gamma-irradiation yielding an absorbed dose of 1.0 Gy, adult CD1 F0 male mice were mated at weekly intervals with unirradiated female CD1 mice. Unirradiated adult males from F1 litters conceived 5 and 6 weeks after paternal F0 irradiation were allowed to mature. Their epididymal spermatozoa were evaluated for in vitro fertilization rates using oocytes from unirradiated 8-12-week-old CD1 females. The mean fertilization rate for spermatozoa from F1 males conceived 5 weeks after paternal F0 irradiation (80.74 +/- 15.74 SD %, n = 5) did not differ significantly from the control fertilization rate (89.40 +/- 10.94 SD %, n = 8). However, the fertilization rate for spermatozoa from F1 males conceived 6 weeks after paternal F0 irradiation (56.14 +/- 21.93 SD %, n = 5) was significantly less than the fertilization rate for control spermatozoa (p < 0.006) or for that of the F1 males conceived 5 weeks after paternal F0 irradiation (p < 0.04). These data suggest that spermatozoa obtained 6 weeks after paternal F0 irradiation can transmit a decrease in fertilization rate to the F1 generation males as well as exhibit decreased fertilization rate themselves when tested directly in vitro. PMID- 9403032 TI - Modulation of NF-kappa B, and Bcl-2 in apoptosis induced by cisplatin in HeLa cells. AB - Cisplatin exposure induces apoptosis in HeLa cells. Since the interaction of this drug with DNA produces reactive oxygen species, we performed an analysis of the oxidative stress-responsive factors AP-1 and NF-kappa B. Although AP-1 levels were not modified during cisplatin exposure, electrophoretic mobility shift assays demonstrated an increase in NF-kappa B DNA binding activity that correlated with a decrease of the inhibitory protein I kappa B alpha and a specific relocalization of c-Rel, as assessed by immunoblotting and immunofluorescence. No changes in the levels or localization of p65 were found. Interestingly, I kappa B alpha relocalized to the nucleus, probably in order to regulate the binding of specific complexes. This process was accompanied by a decrease of the antiapoptotic protein Bcl-2, and a relocalization of p53 protein to the nucleus. Since HeLa cells lost most of their p53 protein due to a specific E6-dependent degradation, cisplatin could be inhibiting this degradation, since the p53 total levels were not increased during the exposure to the drug. PMID- 9403033 TI - Does the increase of 8-hydroxydeoxyguanosine lead to poor sperm quality? AB - Several studies have suggested a population-wide decline in the quality of human semen during the past three decades, but the mechanism remains unclear. It was proved that damage to the DNA of germ cells would lead to mutation, and 8 hydroxydeoxyguanosine (8-OHdG), one of the major products of oxidative damage to DNA, may serve as one of the important factors to affect the sperm quality. In the present paper, 67 semen samples were collected to analyze the sperm quality (density, sperm number, motility, normal head, etc.), and 8-OHdG was measured in DNA isolated from the same sperm samples. The results showed that a significant inverse correlation exists between 8-OHdG/dG(deoxyguanosine) and sperm density (r = -0.358, p = 0.004), and between 8-OHdG/dG and sperm number (r = -0.389, p = 0.01). The results indicate that endogenous oxidative DNA damage could affect sperm quality and subsequently increase the risk of genetic defects. PMID- 9403034 TI - Pilot study of free and conjugated urinary mutagenicity during consumption of pan fried meats: possible modulation by cruciferous vegetables, glutathione S transferase-M1, and N-acetyltransferase-2. AB - Epidemiological and experimental evidence indicates that consumption of fried meats in conjunction with certain genotypes of phase I and II metabolism genes poses an elevated risk for colorectal cancer. Parallel to this, the consumption of cruciferous vegetables is associated with a reduced risk of colon cancer. Therefore, we designed a 6-week pilot feeding study to evaluate the effect of these variables on urinary mutagenicity, which is a biomarker associated with fried-meat consumption. Eight subjects were fed fried meats daily for six weeks; four ate cruciferous vegetables, and four ate non-cruciferous vegetables. Urinary mutagenicity was evaluated in the presence of S9 in strain YG1024 of Salmonella, which is a frameshift strain that overproduces acetyltransferase. C18/methanol extracts of 24-h urines collected once each week were tested unhydrolyzed (free mutagenicity) and hydrolyzed (total mutagenicity); the difference between the two was the conjugated mutagenicity. Although not significant, the levels of conjugated urinary mutagenicity doubled among crucifera consumers and decreased to 30% of the initial levels among non-crucifera consumers, suggesting the possibility that crucifera may enhance the level of conjugated urinary mutagenicity resulting from consumption of fried meats. Such an effect would be consistent with the documented ability of cruciferous vegetables to induce phase II enzymes. The NAT2 rapid phenotype was significantly associated with approximately 2-fold increases in conjugated (p = 0.05) and total (p = 0.004) urinary mutagenicity relative to NAT2 slow subjects, consistent with the elevated risk confirmed by the NAT2 rapid phenotype for colorectal cancer among meat consumers. An approximately 2-fold increase in urinary mutagenicity among the GSTM1- subjects relative to the GSTM1+ subjects approached significance for free (p = 0.18) and total (p = 0.13) urinary mutagenicity. This is the first report on (a) the mutagenicity of hydrolyzed urine, which was consistently more mutagenic than unhydrolyzed urine; (b) the potential enhancement of conjugated urinary mutagenicity by crucifera; and (c) the association of the rapid NAT2 and possibly the GSTM1- phenotype with elevated levels of fried meat-associated urinary mutagenicity. PMID- 9403035 TI - Genetic disorders in house mouse germ cells after the Chernobyl catastrophe. AB - Genetic effects were studied in house mice caught from 1986 to 1994 in regions polluted by radionuclides as a result of the Chernobyl disaster. The dose rates of gamma-radiation on the soil surface ranged from 0.0002 to 2 mGy/h. The frequency of reciprocal translocations in mouse spermatocytes was relatively low, but increased with the dose rate. Embryo mortality was increased only in the progeny of male mice in males caught in 1987 in the area with maximal contamination. The frequency of mice heterozygous for recessive lethal mutations decreased with time after the accident. PMID- 9403036 TI - Frequency of spontaneous and induced micronuclei in the peripheral blood of aging mice. AB - The mouse peripheral blood micronucleus assay, a measure of DNA damage in erythroblastic cells, was used to determine: (1) the incidence of spontaneously occurring micronucleated reticulocytes (MNRETs) as a function of age, and (2) the induction of micronuclei following treatment of young and old animals with mitomycin C. Male C57BL/6 mice, 92 weeks of age, exhibited a significantly higher frequency of spontaneously occurring peripheral blood MNRETs than mice that were 6 or 10 weeks of age. Mice that were 5-6 weeks or 91-92 weeks old were treated with one dose, or two consecutive doses of mitomycin C; this resulted in dose related increases in the frequency of MNRETs. Mitomycin C, at a single dose of 1 or 2 mg/kg, induced one-third as many MNRETs in the older animals as compared to the younger animals. When treated with a split dose of mitomycin C (total dose 0.5 to 2 mg/kg), older animals displayed on average two-thirds the mutagenic response of the younger animals. However, analysis of variance performed on these data indicated that the age of the animals did not have a significant effect on their mutagenic response to mitomycin C at any dose level. It appears that aging mice may not be more sensitive to the mutagenic effects of chemically-induced DNA damage than younger mice, suggesting that the higher spontaneous mutation frequency in older mice could be the result of an increased load of accumulated DNA damage. PMID- 9403037 TI - Involvement of recA and recF in the induced precise excision of Tn10 in Escherichia coli. AB - It has been shown that the increased frequency of precise excision of Tn10 observed after UV or mitomycin C (MMC) treatment or with uvrD- mutants is recA dependent. Previous work has also shown that expression of SOS genes is required for UV- or MMC-induced Tn10 precise excision. In order to determine if the increased excision of Tn10 in uvrD- mutants requires only expression of recA, or expression of other SOS genes, or both, we studied the precise excision of Tn10 in lexA3 (Ind-, SOS non-inducible) and lexA3 recAo98 (operator constitutive recA) mutants. The results of these experiments indicate that the induced excision of Tn10 in the uvrD- null mutant depends on the expression of recA rather than on any of the other genes repressed by LexA. The effect of a null recF mutation on the excision of Tn10 in a uvrD- mutant was also investigated and found to abolish the increased frequencies of this process. Similarly, the recF mutation was found to decrease markedly the increased precise excision of Tn10 induced by MMC in a uvrD+ isogenic strain. These observations indicate that recA and recF are involved in the increased frequencies of Tn10 excision exhibited by uvrD- mutants or after MMC treatment. It remains to be determined whether these two genes participate in these two induction processes in the same biochemical pathways. PMID- 9403038 TI - Proposed mechanism for the photodynamic generation of 8-oxo-7,8-dihydro-2' deoxyguanosine produced in cultured cells by exposure to lomefloxacin. AB - In this study, lomefloxacin (LMX), a widely used quinolone antibiotic with a high frequency of clinical phototoxicity, was investigated by measuring the effects of several antioxidants on its ability to form of 8-oxo-7,8-dihydro-2' deoxyguanosine (8-oxo-dG) in cultured adult rat liver cells after exposure to UVA. In the current study the observed DNA damage, reflected by the formation of 8-oxo-dG, was almost completely inhibited by co-incubation of LMX and cultured cells with sodium azide (NaN3) that specifically quenches singlet oxygen. Vitamin E (alpha-tocopherol), known to quench both superoxide and singlet oxygen, inhibited 8-oxo-dG formation by approximately 54%. Mannitol, a hydroxyl radical scavenger, inhibited 8-oxo-dG formation by 64%. Butylated hydroxyanisole (BHA), a scavenger of hydroxyl, peroxy and alkoxy radicals, showed no inhibition of 8-oxo dG formation but in fact enhanced levels of 8-oxo-dG by 169%. The results of this study suggest that the mechanism for the photodynamic generation of 8-oxo-dG by LMX is mediated, at least in part, by both singlet oxygen and hydroxyl radical and involves both type I and type II photosensitization. PMID- 9403039 TI - Bioactivation of mushroom hydrazines to mutagenic products by mammalian and fungal enzymes. AB - Agaritine (N2-[L-(+)-glutamyl]-4-(hydroxymethylphenyl)hydrazine), the principal hydrazine found in the edible mushroom Agaricus bisporus, as well as the N' acetyl derivative of 4-(hydroxymethyl)phenylhydrazine and 4 (hydroxymethyl)benzene diazonium ion, as the tetraborate salt, considered as the putative proximate and ultimate carcinogens of agaritine, were all synthesised chemically. The mutagenicity of these compounds and of 4-hydrazinobenzoic acid, a precursor of agaritine biosynthesis in mushroom, was investigated in the Ames test, using Salmonella typhimurium strain TA104, in the absence and in the presence of either mushroom tyrosinase or rat hepatic cytosol as activation systems. In the absence of an activation system the diazonium ion was clearly the most mutagenic of the four compounds studied. When tyrosinase was used as activation system, the mutagenicity of N'-acetyl-4-(hydroxymethyl)phenylhydrazine was enhanced; glutathione and superoxide dismutase markedly suppressed the mutagenic response. When the mutagenicity of the four compounds was evaluated in the presence of rat hepatic cytosol, an increase was seen only in the case of N' acetyl-4-(hydroxymethyl)phenylhydrazine; this was shown to be due to deacetylation releasing the more mutagenic free hydrazine. Collectively, the above data are compatible with an activation of agaritine that involves an initial loss of the gamma-glutamyl group followed by microsomal oxidation of the free hydrazine to generate the diazonium ion. Also of interest is the observation that mushroom tyrosinase can convert N'-acetyl-4-(hydroxymethyl)phenylhydrazine to mutagenic product(s); whether these products contribute to the mutagenicity of mushroom extracts remains to be established. PMID- 9403040 TI - Increased sister chromatid exchanges in peripheral lymphocytes of patients with Crohn's disease. AB - A cytogenetic study was performed using Crohn's disease patients to determine whether the presence of chromosome instability is related to Crohn's disease, a chronic inflammatory bowel disease. Sister chromatid exchange (SCE) frequencies in peripheral blood lymphocyte cultures from 22 Crohn's disease patients and an equal number of healthy controls matched for sex and age were analyzed. The mean of SCE frequency in Crohn's disease patients was 11.64 +/- 0.42 (SEM) per cell, which was significantly higher than the value of 8.38 +/- 0.22 per cell in the matched controls (p < 0.0001). The Crohn's disease patients showed significantly increased high frequency cells (HFC) as compared to those among the matched controls. There was a significant correlation between HFC frequencies of the Crohn's disease patients and the severity of their disease as determined by the number of relapses per year and the degree of chronic activity after adjusting for the smoking status (r = 0.54, p = 0.011). In both smokers and non-smokers, the mean SCE and HFC frequencies of the patients were significantly higher than those of the controls. These results suggest that Crohn's disease is a condition with increased chromosome instability characterized by a high level of SCE frequencies which are associated with the inflammatory condition itself. PMID- 9403041 TI - The Janeway Lecture. Iatrogenic carcinogenesis: clinical implications. PMID- 9403042 TI - Intraoperative lymphatic mapping and sentinel lymphadenectomy: community standard care or clinical investigation? PMID- 9403043 TI - Exploring new technologies and treatment strategies for locally advanced prostate cancer. PMID- 9403044 TI - Positron emission tomography (PET)--measured biochemical response to radiotherapy of laryngeal tumors. PMID- 9403045 TI - Sentinel lymphadenectomy for breast cancer in a community managed care setting. AB - PURPOSE: To evaluate the feasibility, accuracy, and reproducibility of intraoperative lymphatic mapping and sentinel lymphadenectomy (IOLM/SL) in the staging of breast cancer patients in a community managed care setting. PATIENTS AND METHODS: One hundred forty-five patients with primary breast cancer were prospectively studied over a 26-month period. They underwent vital dye injection at their primary breast cancer site. Lymphatic channels were traced to the sentinel lymph node, which was excised, serially sectioned, and examined. A level I and II axillary lymph node dissection and definitive breast surgery were then performed. RESULTS: Sentinel nodes were identified in 103 of 145 procedures (71.0%). Sentinel and nonsentinel lymph nodes were concordant in 100 of 103 cases (97.1%). Three patients (9.7%) had falsely negative sentinel nodes; there were none in the last 80 patients. Of 28 positive sentinel nodes, 12 (42.9%) represented the only tumor-containing node within the axilla. Sentinel nodes were significantly more likely to contain tumor than nonsentinel nodes (33/50, 66.0% vs 54/467, 11.6%, P < 0.0001). IOLM/SL identified more micrometastases (< 2 mm) than standard axillary lymph node dissection (13/33, 39.6% vs 4/177, 2.2%, P < 0.001). Nine of 42 patients (21.4%) whose sentinel node could not be identified had five or more nodal metastases. Two of six patients with presumed Tis primaries had nodal metastases. DISCUSSION: IOLM/SL accurately identifies the sentinel lymph node(s) most likely to contain metastatic disease. A procedural learning curve was present. An unsuccessful IOLM/SL was a risk factor for considerable nodal metastases. IOLM/SL with a tumor-free sentinel node may obviate a formal axillary lymph node dissection. The technique was feasible, economical, and reproducible within the context of a community managed care facility, while not placing exacting demands on operating room, pathology, or nuclear medicine personnel. PMID- 9403046 TI - Radioguided surgery for the ultrastaging of the patient with melanoma. AB - PURPOSE: Lymphatic mapping techniques have changed the standard of surgical care for the malignant melanoma population and are being investigated to improve the staging and decrease the morbidity of patients with all types of cancer. This study aimed to describe a combination of techniques and the use of multiple disciplines for accurately staging and treating patients with melanoma. MATERIALS AND METHODS: Over a 4-year period, 595 patients were studied using a protocol consisting of preoperative lymphoscintigraphy using filtered technetium sulfur colloid to define all regional basins at risk for metastatic disease, and intraoperative lymphatic mapping with a vital blue dye and radiocolloid to identify the node in the basin most at risk for metastases (the sentinel lymph node). Detailed pathological exam (serial sectioning, immunohistochemical staining, reverse transcriptase polymerase chain reaction [RT-PCR] analysis) of the sentinel lymph node was used to stage the melanoma patient. RESULTS: A combination of blue dye and radiocolloid intraoperative mapping resulted in a 98% success rate for the identification of the sentinel lymph node. Routine pathological examination identified 73.8% of the metastases. The remainder were detected with serial sectioning (7.8%) and immunohistochemical staining (18.4%). RT-PCR analysis based on a tyrosinase probe has upstaged 47% of the histologic sentinel lymph node-negative population. CONCLUSION: Lymphatic mapping technology provides accurate staging of the melanoma patient, at lower costs for the health care system and a lower morbidity for the patient. PMID- 9403047 TI - Conformal high dose rate iridium-192 boost brachytherapy in locally advanced prostate cancer: superior prostate-specific antigen response compared with external beam treatment. AB - PURPOSE: Prostate-specific antigen levels are used to judge disease control of prostate cancer. No published data attest to the greater ability of conformal brachytherapy to control disease compared with conventional radiation at a single institution. This report compares the biochemical response rates in patients with stages T2b to T3c prostate cancer treated with conformal brachytherapy boost and external beam radiation with the rates in patients treated with conventional external radiation alone. MATERIALS AND METHODS: From November 1991 through November 1995, 58 patients received 45.6 Gy pelvic external irradiation and three high dose rate iridium-192 conformal boost implants of 5.5 to 6.5 Gy each. They were compared with 278 similarly staged patients treated from January 1987 through December 1991 with external beam radiation to prostate-only fields (median dose 66.6 Gy). No patient received androgen deprivation. Patient outcome was analyzed for biochemical control. Biochemical failure was defined as a prostate-specific antigen level > 1.5 ng/mL and rising on two consecutive values. If serial posttreatment prostate-specific antigen levels were showing a continuous downward trend, failure was not scored. RESULTS: Median follow-up was 43 months for the conventionally treated group and 26 months for the brachytherapy boost group. The median pretreatment prostate-specific antigen level was 14.3 ng/mL for the external-beam-radiation-alone group and 14.0 ng/mL for the brachytherapy boost group. The median Gleason scores were 6 and 7, respectively, for the two groups. The biochemical control rate was significantly higher in the brachytherapy boost treatment group. Three-year actuarial biochemical control rates were 85% versus 52% for the conformally and conventionally treated patients, respectively. In a multivariate analysis, the use of conformal brachytherapy boost and pretreatment prostate-specific antigen level were significant prognostic determinants of biochemical control. The 3-year actuarial rates of biochemical control for conformally versus conventionally treated patients, respectively, were 83% versus 72% for a pretreatment prostate specific antigen level of 4.1 to 10.0 ng/mL, 85% versus 47% for a prostate specific antigen level of 10.1 to 20.0 ng/mL, and 89% versus 29% for prostate specific antigen > 20 ng/mL. When the analysis was limited to patients in both groups with a minimum 12-month follow-up, the brachytherapy boost group continued to show a higher biochemical control rate compared with the conventional radiation group (3-year actuarial rates of 86% vs 53%). DISCUSSION: These preliminary results show a significant improvement in the biochemical response rate with conformal boost brachytherapy and pelvic external radiation compared with conventional radiation alone. These results, coupled with our previously reported acceptable toxicity rates, support the use of this technique. PMID- 9403048 TI - Can positron emission tomography distinguish tumor recurrence from irradiation sequelae in patients treated for larynx cancer? AB - PURPOSE: Distinguishing persistent or recurrent tumor from post-radiation edema or soft-tissue/cartilage necrosis in patients treated for carcinoma of the larynx can be difficult. Because recurrent tumor is often submucosal, multiple deep biopsies may be necessary before a diagnosis can be established. Positron emission tomography with F-18 fluorodeoxyglucose was studied for its ability to aid in this problem. PATIENTS AND METHODS: FDG PET scans were performed on 31 patients who were suspected of having persistent or recurrent tumor after radiation treatment for carcinoma of the larynx. Patients underwent thorough history and physical examinations, scans with computed tomography (23 patients), and pathological evaluation when indicated. PET scans were interpreted by each of the two radiologists, who were blinded to patient outcome and the other's report. RESULTS: The time between completion of radiation treatment and positron emission tomography examination ranged from 2 to 61 months with a median of 6 months. Fifteen patients had pathological evidence of tumor in the larynx, while 16 have remained without evidence of disease. The overall sensitivity and specificity of the positron emission tomography interpretations were 80% and 81%, respectively. The sensitivity and specificity of the computed tomography scan interpretations were 58% and 100%, respectively. Of the 23 patients with computed tomography scans, eight patients acquired useful information from the positron emission tomography, three patients had incorrect positron emission tomography interpretations and correct computed tomography interpretations, and one patient had positive tumor despite a negative positron emission tomography and computed tomography. DISCUSSION: Positron emission tomography is useful in distinguishing benign from malignant changes in the larynx after radiation treatment. This noninvasive technique can supplement information provided by computed tomography scans. It is reasonable to delay biopsy, which could traumatize radiation-damaged tissues and precipitate necrosis, for those patients with negative positron emission tomography scans who have clinical signs and symptoms associated with recurrence. PMID- 9403049 TI - Whole-abdomen radiation therapy as salvage treatment for epithelial ovarian carcinoma. AB - PURPOSE: This study aimed to evaluate the efficacy and safety of whole-abdomen radiation therapy as salvage treatment in patients with ovarian cancer. PATIENTS AND METHODS: Twenty-seven patients who failed aggressive cytoreductive surgery followed by multiple-drug platinum-based chemotherapy were found to have recurrent epithelial carcinoma of the ovary and were treated with whole-abdomen radiation as salvage therapy. Dosage fractions were planned at 100 to 150 cGy daily to 3000 to 3500 cGy, followed by a pelvic boost at 150 to 180 cGy daily. All patients completed the planned treatment. The average treatment program required 53.5 days (range, 42-71 days). RESULTS: Survival rates at years 1 through 5 were 66%, 48%, 26%, 15%, and 15%, respectively. Residual disease at initiation of radiation correlated strongly with length of survival. The patients with microscopic disease survived an average of 63 months (range, 30-111 months). Patients with disease larger than 2 cm survived an average of 9 months (range, 5 17 months). Toxicity was seen in all patients. Eight patients experienced grade 3 or 4 toxicity, primarily white blood cell count and gastrointestinal toxicity. There were no deaths related to toxicity. DISCUSSION: This experience strongly suggests that whole-abdomen radiation is a viable salvage option, especially for patients with microscopic retroperitoneal disease or small-volume macroscopic disease. PMID- 9403051 TI - Telomerase. The end-replication problem. PMID- 9403050 TI - TA90 immune complex predicts survival following surgery and adjuvant vaccine immunotherapy for stage IV melanoma. AB - PURPOSE: Although prognosis remains poor for patients with distant metastatic melanoma, we have observed significantly prolonged survival in patients receiving our polyvalent melanoma cell vaccine (PMCV) following complete metastasectomy for American Joint Committee on Cancer (AJCC) stage IV melanoma. Clinical prognostic factors specific to this stage IV subgroup have not been well characterized. We previously reported that the serum immune complex (IC) level of a 90-kD glycoprotein antigen (TA90) was an objective predictor of survival and recurrence in patients with early-stage melanoma. In the present study we correlated the postoperative TA90-IC level of AJCC stage IV patients prior to adjuvant PMCV therapy with their duration of subsequent survival. PATIENTS AND METHODS: From October 1, 1984, to December 31, 1995, 125 stage IV patients began PMCV after complete resection of distant melanoma metastases. One blood sample was obtained immediately prior to vaccine therapy, and the serum TA90-IC level was assessed as positive or negative using our double-determinant ELISA. Disease-free and overall survival were recorded prospectively from the start of vaccine therapy. The correlation between prevaccine TA90-IC level and survival was assessed by the log rank test and Cox proportional hazards model. RESULTS: Median follow-up after PMCV therapy was 36.5 months, with a minimum of 12 months. Univariate analysis demonstrated that TA90-IC level is significant for both overall survival and disease-free survival. Median overall survival, median disease-free survival, and rate of 5-year survival were higher for patients with negative TA90-IC levels than for those with positive TA90-IC level (58 vs 19 months, 7 vs 4 months, and 49% vs 27%, respectively). Multivariate analysis established TA90-IC as an independent prognostic indicator for both overall and disease-free survival following adjuvant PMCV therapy for AJCC stage IV melanoma. CONCLUSION: Prevaccine TA90-IC level correlated strongly with overall and disease-free survival in our stage IV melanoma patients receiving postoperative PMCV immunotherapy. This is the first serum marker shown to have importance in predicting the survival of melanoma patients receiving adjuvant immunotherapy after complete resection of distant metastases. PMID- 9403052 TI - Mapping quantitative trait loci for fear-like behaviors in mice. AB - Two mouse models developed for screening anxiolytic drugs were selected for genetic analysis, namely "wall-seeking" tendency in an open field ("thigmotaxis") and the light-to-dark transition (LD) paradigm, a conflict test. These tests measure differences in naturalistic tendencies of mice to explore a novel environment and to avoid a bright light or the center of an open field. In an F2 intercross of two strains of mice (A/J and C57BL/6J) that differ markedly in these behaviors, we estimated a broad sense heritability ranging from 0.3 to 0.59. With this intercross (n = 518), we have mapped several quantitative trait loci (QTL) for these behaviors by performing a genome-wide search. A significant QTL on chromosome 10 (near D10Mit237; LOD of 9.3) that affects LD behavior was identified, and suggestive QTL (LOD > 2.8) were mapped to chromosomes 6, 15, 19, and X. For center time behaviors, QTL were identified on chromosome 1 (LOD of 7.7 and 4.0 for the initial 5-min epoch and the first trial average of the next two 5 min epochs, respectively), and suggestive QTL (LOD > 2.8) were mapped to chromosomes 6 and 14. These QTL individually explain from 2.3 to 8.4% of the phenotypic variance. Collectively, the multiple independent QTL explain from 3.5 to 26.5% of the F2 population's phenotypic variance, depending on the trait. The complexity and heterogeneity of the genetic factors underlying these fear-like behaviors are illustrated by the lack of shared QTL between paradigms and by mapping different QTL for repeated trials of behavior. The identification of QTL affecting individual differences in fear-like behavior may lead to the identification of new gene products and pathways that modulate behavior, providing targets for rational drug design. PMID- 9403053 TI - A 2.5-Mb transcript map of a tumor-suppressing subchromosomal transferable fragment from 11p15.5, and isolation and sequence analysis of three novel genes. AB - 11p15.5 is an important tumor-suppressor gene region, showing loss of heterozygosity in Wilms tumor, rhabdomyosarcoma, adrenocortical carcinoma, and lung, ovarian, and breast cancer. We previously mapped directly by genetic complementation a subtransferable fragment (STF) harboring an embryonal tumor suppressor gene and spanning about 2.5 Mb. We have now mapped the centromeric end of this STF between D11S988 and D11S12 and its telomeric end between D11S1318 and TH. We have isolated a complete contig of PAC, P1, BAC, and cosmid genomic clones spanning the entire 2.5-Mb region defined by this STF, as well as more than 200 exons from these genomic clones using exon trapping. We have isolated genes in this region by directly screening DNA libraries as well as by database searching for ESTs. Nine of these genes have been reported previously by us and by others. However, the initial mapping of most of those genes was based on FISH or somatic cell hybrid analysis, and here we precisely define their physical location. These genes include RRM1, GOK (D11S4896E), Nup98, CARS, hNAP2 (NAP1L4), p57KIP2 (CDKN1C), KVLQT1 (KCNA9), TAPA-1, and ASCL2. In addition, we have identified several novel genes in this region, three of which, termed TSSC1, TSSC2, and TSSC3, are reported here. TSSC1 shows homology to Rb-associated protein p48 and chromatin assembly factor CAF1, and it is located between GOK and Nup98. TSSC2 is homologous to Caenorhabditis elegans beta-mannosyl transferase, and it lies between Nup98 and CARS. TSSC3 shows homology to mouse TDAG51, which is implicated in FasL-mediated apoptosis, and it is located between hNAP2 and p57KIP2. Thus, these genes may play a role in malignancies that involve this region. PMID- 9403054 TI - Improved analysis of microsatellites using mass spectrometry. AB - The primer oligo base extension reaction combined with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, recently introduced by our group for detection of single-point mutations and small insertions/deletions, has been applied to the reliable quantification of nucleotide repeat units in microsatellites. The AluVpA DNA marker within intron 5 of the interferon-alpha receptor gene was chosen as the model system. By varying the dNTP/ddNTP mixtures used, the assay could also be directed to detect the location of second-site mutations within the repeats, resulting in identification of alleles not detectable by electrophoretic sizing methods and thus an increase of the polymorphism information content for a sampling of 28 unrelated individuals. The method results in highly informative mass signals and has the potential to increase the polymorphism information content for systems containing second-site mutations; thus it is a very attractive alternative technique in statistics-based gene mapping, cancer diagnostics, and forensic applications. PMID- 9403055 TI - Comparison of DNA sequences with protein sequences. AB - The FASTA package of sequence comparison programs has been expanded to include FASTX and FASTY, which compare a DNA sequence to a protein sequence database, translating the DNA sequence in three frames and aligning the translated DNA sequence to each sequence in the protein database, allowing gaps and frameshifts. Also new are TFASTX and TFASTY, which compare a protein sequence to a DNA sequence database, translating each sequence in the DNA database in six frames and scoring alignments with gaps and frameshifts. FASTX and TFASTX allow only frameshifts between codons, while FASTY and TFASTY allow substitutions or frameshifts within a codon. We examined the performance of FASTX and FASTY using different gap-opening, gap-extension, frameshift, and nucleotide substitution penalties. In general, FASTX and FASTY perform equivalently when query sequences contain 0-10% errors. We also evaluated the statistical estimates reported by FASTX and FASTY. These estimates are quite accurate, except when an out-of-frame translation produces a low-complexity protein sequence. We used FASTX to scan the Mycoplasma genitalium, Haemophilus influenzae, and Methanococcus jannaschii genomes for unidentified or misidentified protein-coding genes. We found at least 9 new protein-coding genes in the three genomes and at least 35 genes with potentially incorrect boundaries. PMID- 9403056 TI - A tool for analyzing and annotating genomic sequences. AB - We describe a tool for analyzing and annotating large genomic sequences containing introns. The analysis and annotation tool (AAT) includes two sets of programs, one for comparing the query sequence with a protein database and the other for comparing the query with a cDNA database. Each set contains a fast database search program and a rigorous alignment program. The database search program quickly identifies regions of the query sequence that are similar to a database sequence. Then the alignment program constructs an optimal alignment for each region and the database sequence. The alignment program also reports the coordinates of exons in the query sequence. Pairwise alignments of the query sequence with protein and cDNA database sequences are combined into multiple sequence alignments, which provide a view of all protein and cDNA sequences matching a query region. On a data set of 570 DNA sequences, AAT identified 94% of coding nucleotides correctly and 74% of exons exactly. Results of analyzing a human BAC sequence with the AAT tool are also presented. The AAT tool reduces the labor-intensive work of locating the exons of the query sequence and improves the process of defining intron-exon boundaries by using the wealth of available protein and cDNA data. PMID- 9403057 TI - Comparative gene mapping of the human and mouse TEP1 genes, which encode one protein component of telomerases. AB - The chromosomal locations of the human TEP1 (telomerase protein component 1) and mouse Tep1 genes, which were originally named TLP1 (telomerase protein 1) or TP1 (telomerase-associated protein 1), were determined by direct R-banding FISH and a molecular linkage analysis with interspecific backcross mice. The human TEP1 and mouse Tep1 genes were mapped by FISH to human chromosome 14q11.2 and to the C2D1 band of mouse chromosome 14, respectively. By means of genetic linkage mapping, the mouse gene was further localized as being 2.7 cM distal to D14Mit18 and D14Mit134 and 2.0 cM proximal to D14Mit5 on mouse chromosome 14, where conserved linkage homology with human chromosome 14q11-q12 has been identified. PMID- 9403058 TI - The telomere-associated DNA from human chromosome 20p contains a pseudotelomere structure and shares sequences with the subtelomeric regions of 4q and 18p. AB - The human chromosome 20p telomere has been cloned on a yeast artificial chromosome (YAC). The telomere-associated DNA contains an interstitial tract of (TTAGGG)n telomeric repeats 60 kb in from the chromosome end. Frequent truncation of the YAC clone was observed due to resolution of the internal telomeric array into a telomere. The 20p internal telomeric repeat tract is flanked on its centromeric side by telomere-associated repeated sequences that have previously been found adjacent to terminal telomeric repeat arrays. The pseudotelomere structure of the 20p subtelomeric region is similar to the structure of some yeast subtelomeric regions where these sequences act as substrates for recombinational repair of chromosome ends that have lost their terminal telomeric repeat arrays. Sequences flanking the telomeric end of the internal (TTAGGG)n repeat array on 20p are found adjacent to three other subtelomeric (TTAGGG)n tracts on 4q, 18p, and an unknown chromosome end, respectively. These shared sequences provide evidence of exchange between nonhomologous chromosomes in humans. PMID- 9403059 TI - The Charcot-Marie-Tooth binary repeat contains a gene transcribed from the opposite strand of a partially duplicated region of the COX10 gene. AB - Misalignment between the two elements of the CMT1A-REP binary repeat on chromosome 17p11.2-p12 causes two inherited peripheral neuropathies, Charcot Marie-Tooth type 1A (CMT1A) and hereditary neuropathy with liability to pressure palsies. This binary repeat contains repetitive DNA elements that include LINES, SINES, medium reiteration frequency repeats, and a transposon-like element. The COX10 gene has been mapped 10 kb centromeric to the distal CMT1A-REP element, and a portion of this gene is present in both the proximal and the distal CMT1A-REP elements. We report the isolation and characterization of a novel cDNA (C170RF1), which maps centromeric to and partially within the proximal CMT1A-REP element. Part of C170RF1 is transcribed from the opposite strand of the COX10 partial gene duplication present in the proximal CMT1A-REP element. This finding shows that C170RF1 and COX10 are being transcribed from opposite strands of identical DNA sequences that are separated by 1.5 Mb in the genome. RT-PCR analysis showed the proximal transcript was expressed in skeletal muscle. Sequence analysis identified an open reading frame encoding a 199-amino-acid protein. Zoo blot analysis showed that the transcript is conserved in nonhuman primates. The presence of a binary repeat contributes to the instability of this region of chromosome 17, yet two CMT1A-REP elements are present in the chimpanzee and all human populations. The presence of expressed sequences in both elements of the CMT1A-REP binary repeat could explain the maintenance of this repeat in humans. PMID- 9403060 TI - Identification of new translocation breakpoints at 12q13 in lipomas. AB - Cytogenetic studies of banded chromosomes and fluorescence in situ hybridization (FISH) of several yeast artificial chromosomes (YACs) that are part of a 128-kb resolution physical map of a portion of 12q13 revealed that 4/14 (28%) lipomas have breakpoints in 12q13. These breakpoints are more than 10 Mb away from the HMGIC gene at 12q14-q15, which is known to be modified in some lipomas. FISH with individual YACs at 12q13 enabled us to identify four YACs that span three breakpoints. Our results suggest that genes other than HMGIC on human chromosome 12 may be involved in the etiology of lipoma development. PMID- 9403061 TI - Cloning and characterization of freac-9 (FKHL17), a novel kidney-expressed human forkhead gene that maps to chromosome 1p32-p34. AB - We describe the cloning of a near full-length cDNA of 4258 nucleotides encoding freac-9 (HGMW-approved symbol FKHL17), a novel human forkhead gene. The 5' untranslated region is unusual since it is very long, 2127 nucleotides, and contains 15 upstream AUG codons. Hybridization to a panel consisting of RNA derived from 50 different tissues showed that freac-9 is transcribed exclusively in the kidney. The kidney-derived cell lines COS-7 and 293 are shown to express freac-9. A combination of fluorescence in situ hybridization and somatic cell hybrids localizes freac-9 to the chromosomal region of 1p32-p34. The conceptual translation product predicts a protein of 372 amino acids with an N-terminal domain rich in acidic amino acids and with a high likelihood of forming an amphipatic helix, a DNA binding forkhead domain, and a C-terminal region that has a high probability of forming an amphipatic beta-sheet. The amino acid sequence of the DNA binding forkhead motif of FREAC-9 is identical to that of another forkhead protein, FREAC-4, whereas 12 substitutions are present at the nucleotide level. There are no similarities in regions outside of the DNA binding domains of FREAC-9 and FREAC-4 and since freac-4 maps to a different chromosome (5q12-q13) it is likely that an evolutionary selection has acted to maintain identical DNA binding domains between these two kidney expressed transcription factors. PMID- 9403062 TI - Aberrant mRNA splicing causes sorbitol dehydrogenase deficiency in C57BL/LiA mice. AB - Sorbitol dehydrogenase (Sord) catalyzes the interconversion of sorbitol and fructose and is functionally important both in the metabolism of dietary sorbitol and as a source of fructose in semen. Together with aldose reductase, Sord forms the polyol pathway, which plays an important role in the etiology of diabetic complications. The Sord-deficient mouse (C57BL/ LiA) is very useful in animal model studies of the involvement of the polyol pathway in both diabetic and congenital cataracts. To understand more about this strain, we characterized the molecular basis underlying this Sord deficiency and found that this was due to a point mutation in the exon 8/intron 8 junction. Substitution of an A for G at the first position of the strictly conserved GT donor completely abolished normal splicing of exon 8. Aberrant splicing of this junction generates at least three types of transcripts: one lacking exon 8, another that has a truncated exon 8, and a third that contains intron sequences. We have devised two convenient PCR based methods to identify this mutation in C57BL/LiA mice. These methods are useful in animal experiments that involve cross-breeding with these mice because they allow early determination of genotype without the need to sacrifice the animals for enzyme assay. PMID- 9403063 TI - Genomic structure and chromosomal mapping of the nuclear orphan receptor ROR gamma (RORC) gene. AB - The nuclear orphan receptor subfamily ROR/RZR is part of the steroid and thyroid hormone/retinoid receptor superfamily and consists of three different genes, alpha, beta, and gamma. In this study, we determined the genomic structure of mouse ROR gamma and the chromosomal localization of both mouse ROR gamma and human ROR gamma (HGMW-approved symbol RORC). The genomic structure of the mouse ROR gamma gene was derived from the analysis of P1 vector clones containing large genomic fragments encoding ROR gamma. These results revealed that the mROR gamma gene has a complex structure consisting of 11 exons separated by 10 introns spanning more than 21 kb of genomic DNA. The DNA-binding domain is contained in two exons, 3 and 4, each encoding one zinc-finger. The splice site between exon 3 and exon 4 is identical to that found in RAR and TR3 receptors. ROR gamma is expressed as two mRNAs, 2.3 and 3.0 kb in size, that are derived by the use of alternative polyadenylation signals. We show by fluorescence in situ hybridization that the mouse ROR gamma gene is located on chromosome 3, in a region that corresponds to band 3F2.1-2.2. The human ROR gamma was mapped to chromosome region 1q21. The results demonstrate that the ROR gamma genes are located in chromosomal regions that are syntenic between mouse and human. PMID- 9403064 TI - Characterization of human homologs of the Drosophila seven in absentia (sina) gene. AB - Studies of Drosophila photoreceptor development have illustrated the means by which signal transduction events regulate cell fate decisions in a multicellular organization. Development of the R7 photoreceptor is best understood, and its formation is dependent on the seven in absentia (sina) gene. We have characterized two highly conserved human homologs of sina, termed SIAH1 and SIAH2. SIAH1 maps to chromosome 16q12 and encodes a 282-amino-acid protein with 76% amino acid identity to the Drosophila SINA protein. SIAH2 maps to chromosome 3q25 and encodes a 324-amino-acid protein that shares 68% identity with Drosophila SINA and 77% identity with human SIAH1. SIAH1 and SIAH2 were expressed in many normal and neoplastic tissues, and only subtle differences in their expression were noted. However, one of three murine homologs, Siah1B, was strongly induced in fibroblasts undergoing apoptotic cell death. While a previous study suggested that SINA was a nuclear protein, epitope-tagged SINA and SIAH1 proteins were found in the cytoplasm of Drosophila and mammalian cells. Their substantial evolutionary conservation, role in specifying cell fate, and activation in apoptotic cells suggest the SIAH proteins have important roles in vertebrate development. Furthermore, given the role of sina in Drosophila photoreceptor development, SIAH2 is a candidate for the Usher syndrome type 3 gene at chromosome 3q21-q25. PMID- 9403065 TI - Molecular cloning and characterization of the human mitochondrial 2,4-dienoyl-CoA reductase gene (DECR). AB - 2,4-Dienoyl-CoA reductase (EC 1.3.1.34) is an auxiliary enzyme of beta-oxidation, and it participates in the metabolism of unsaturated fatty enoyl-CoA esters having double bonds in both even- and odd-numbered positions. In this article we describe the molecular cloning of the human gene for the 120-kDa isoform of mitochondrial 2,4-dienoyl-CoA reductase (DECR). The gene is approximately 30 kb and comprises 10 exons varying in size from 79 to 203 bp and 9 introns whose sizes vary from 95 bp to about 10 kb. The 5' UTR and 3' UTR are included in exons 1 and 10, respectively. The promoter region contains putative binding sites for several transcription factors, e.g., Sp1, AP-2, AP-4, and C/EBP, but no TATA box was found. Primer extension analysis and 5' RACE-PCR revealed variability in the length of the 5'-UTR, the longest being 72 bp. Through the use of FISH analysis on metaphase chromosomes with a genomic fragment of 2,4-dienoyl-CoA reductase, the gene was assigned to the chromosomal band 8q21.3. PMID- 9403066 TI - Mouse connexin40: gene structure and promoter analysis. AB - A family of related connexin genes encodes the subunit gap junction proteins that form intercellular channels in different tissues. Connexin40 (Cx40) is one of these proteins, and it exhibits limited expression only in a few cells of the cardiovascular system. To begin to analyze Cx40 expression, we isolated a 3.3-kb rat Cx40 cDNA by hybridization screening of a bacteriophage library prepared from BWEM cells and isolated corresponding mouse genomic clones from a bacterial artificial chromosome library. Restriction mapping, sequencing, and comparison to the rat cDNA showed that the mouse Cx40 gene contained a short first exon, an 11.4-kb intron, and a second exon containing the complete coding region and 3' UTR. Exon I contained only 1 base that differed between rat and mouse. Primer extension experiments yielded a single band and confirmed the position of the transcriptional start site. We obtained 1.2 kb of sequence 5' of the transcriptional start site and 400 bp 3' of exon I. Exon I was closely preceded by a consensus TATA box. The flanking sequences contained a number of potential transcription factor binding sites (including AP-1, AP-2, SP1, TRE, and p53). To identify transcriptional regulatory elements in the Cx40 promoter region, a series of DNA deletion fragments flanking exon I was prepared, subcloned adjacent to a luciferase reporter gene, and used for transient transfections of BWEM, SHM, and N2A cells. The resulting luciferase activity determinations suggested that an area of 300 bp 5' of the transcription start site acted as a basal promoter for Cx40 and that there was a strong negative regulatory element in the region from +100 to +297. PMID- 9403067 TI - Site-specific retrotransposition of L1 elements within human alphoid satellite sequences. AB - In the course of a search for microsatellites as centromeric polymorphic markers at the 3' ends of Alu or L1 elements, we observed a much higher frequency of L1 than Alu elements embedded within alpha satellite DNA. By sequence analysis of the L1 elements at their alphoid locus of insertion, we found that the insertion site was specific, with the consensus being (Py)2-10/ (Pu)3-7. All potential sites within the consensus alphoid 171-bp repeat are occupied by such elements. This confirms the finding by Feng et al. (1996; Human retrotransposon encodes a conserved endonuclease required for retrotransposition, Cell 87:905-916) that the progenitor L1 elements encode a site-specific endonuclease and that they generate copies that are inserted at these specific sites. The analysis of retrotransposed L1 elements within the alphoid domains of the acrocentric chromosomes showed that a number of loci are shared among all five acrocentrics. This sheds light on the manner in which centromeric regions of these chromosomes are exchanging information during evolution. PMID- 9403068 TI - Cloning and chromosomal mapping of human casein kinase I gamma 2 (CSNK1G2). AB - A clone of immature cDNA for human casein kinase I gamma 2 (CSNK1G2) was isolated by screening the human testis cDNA library with a PCR-amplified probe (about 400 bp) representing the kinase domain of rat casein kinase I gamma 2 (CKI gamma 2). Comparison of the entire sequence with that of rat CKI gamma 2 showed that the cDNA contained the complete coding sequence of CKI gamma 2 as well as an intron like sequence of 1006 bp, part of which was homologous to the Alu sequence. To obtain an insertion-free CSNK1G2 cDNA, PCR cloning was performed based on the above sequence. The amplified 1687-bp fragment was subcloned and sequenced. The predicted amino acid sequence consisted of 416 residues, 94% of which were identical to that of the rat homologue. Although there are two Src homology 3 (SH3) domain-binding motifs (Pro-X-X-Pro consensus), Pro-Lys-Val-Pro and Pro-Ser Glu-Pro in the C-terminal region of rat CKI gamma 2, only the latter was conserved in the human counterpart. This finding suggests that the latter motif is important for binding to the signal transduction adaptor protein Nck (NCK). The human CSNK1G2 gene was mapped to chromosome 19p13.3 by fluorescence in situ hybridization and PCR analysis of the human/rodent hybrid cell panel. PMID- 9403069 TI - Mapping of eight testis-specific genes to mouse chromosomes. AB - We previously identified eight testis-specific genes using antibodies raised against testicular germ cells. They are expressed during spermatogenesis and are presumed to be involved in testicular germ cell differentiation and sperm formation. We have mapped the genomic loci for these testis-specific genes using restriction fragment length variants in interspecific backcross mice. The calmegin gene (Clgn) was mapped to Chr 8. The synaptonemal complex protein gene 1 (Sycp1) probe hybridized with two sequences on different chromosomes; Sycp1-rs2 was mapped to Chr 3, whereas Sycp1-rs3 was mapped to Chr 7. The relaxin-like factor gene (Rlnl) was mapped to Chr 8, and collapsin response mediator protein 1 (Crmp1) was mapped to Chr 5. Three novel genes encoding testis-specific proteins A2 (Tsga2), A8 (Tsga8), and A12 (Tsga12) were mapped to chromosomes 3, X, and 10, respectively. PMID- 9403070 TI - Use of the Indian muntjac idiogram to align conserved chromosomal segments in sheep and human genomes by chromosome painting. AB - We have hybridized all 28 chromosome-specific painting probes from the domestic sheep (Ovis aries, 2n = 54) onto metaphase chromosomes of the Indian muntjac deer (Muntiacus muntjak vaginalis, 2n = 6,7) and identified 35 conserved chromosomal segments. Results from this study show that most of the sheep acrocentric chromosomes hybridized to single regions in the Indian muntjac genome. This conserved hybridization pattern supports the concept that the large Indian muntjac chromosomes were derived from multiple tandem fusions from an ancestral deer species. Using previously reported fluorescence in situ hybridization data in which human chromosomes were hybridized onto the Indian muntjac genome, we were able to align chromosomal segments of the sheep and human genomes. Using this three-species genome alignment approach, we have identified a minimum of 42 conserved chromosomal segments between sheep and human genomes including 7 new regions not previously reported. PMID- 9403071 TI - Mapping of the human ribosomal large subunit protein gene RPL29 to human chromosome 3q29-qter. AB - The human ribosomal protein L29, which we reported previously, was subsequently shown to have the same nucleotide sequence as that of cell surface heparin/heparan sulfate-binding protein, designated HP/HS interacting protein. A polymerase chain reaction-based strategy was used to distinguish the functional intron-containing gene RPL29 (HGMW-approved symbol) from multiple pseudogenes. By somatic cell hybrid analysis, radiation hybrid mapping, and fluorescence in situ hybridization, we have located RPL29 on the telomeric region of the q arm of chromosome 3. RPL29 is the most distal marker of the long armof chromosome 3. Of the human ribosomal protein genes mapped, RPL29 is the shortest distance from another ribosomal protein gene marker, RPL35 a which has also been mapped to the 3q29-qter region. PMID- 9403072 TI - Linkage mapping of Thiel-Behnke corneal dystrophy (CDB2) to chromosome 10q23-q24. AB - Corneal dystrophy of the anterior basement membrane is a heterogeneous set of diseases characterized by painful, recurrent, bilateral erosions of the cornea, which often result in significant visual impairment. There are several similar but clinically distinct forms of anterior basement membrane/Bowman's membrane disease, including two autosomal dominant forms, Reis-Bucklers and Thiel-Behnke corneal dystrophy. Genes causing autosomal, nonsyndromic corneal dystrophy have been mapped to human chromosomes 1p, 5q, 12q, 16q, 17p, and 20p. Using microsatellite markers closely linked to the known corneal dystrophy loci, we excluded linkage between the known sites and the disease locus in a large, four generation family with Thiel-Behnke corneal dystrophy. A genome-wide search using a panel of microsatellite markers demonstrated a maximum two-point lod score of 4.0 at 0% recombination between the disease locus in this family and the marker D10S1239, which maps to 10q23-q24. Testing with additional microsatellite markers from 10q places the disease locus between D10S677 and D10S1671, a distance of approximately 12.0 cM, with a maximum multipoint lod score of 5.5. Based on this evidence, we have identified another locus (CDB2) for corneal dystrophy of the anterior basement membrane/Bowman's membrane, Thiel-Behnke type, further demonstrating the exceptional genetic and phenotypic heterogeneity of these diseases. PMID- 9403073 TI - The chromosome location of the human homolog of the mouse mammary tumor associated gene INT6 and its status in human breast carcinomas. AB - The INT6 gene is a common integration site for the mouse mammary tumor virus in mouse mammary tumors. We have determined that the human homolog of INT6 is located on chromosome region 8q22-q23. A processed INT6 pseudogene is located on chromosome 6q. INT6 is composed of 13 exons that span 45 kb of genomic DNA. The deduced amino acid sequence of the gene product is identical to the mouse protein and contains three potential translation start signals. We have examined 100 primary breast carcinoma DNAs for evidence of genetic alteration affecting INT6. Loss of heterozyosity (LOH) was detected in 11 of 39 (28%) of the tumor samples informative for a polymorphic sequence in intron 7 of INT6. Since single-strand conformation and hybrid mismatch analysis of the remaining allele in these tumor DNAs failed to detect any mutations, we conclude that the target gene for LOH must be closely linked to INT6. PMID- 9403074 TI - Evidence for linkage of chromosome 12q15-q24.1 markers to high total serum IgE concentrations in children of the German Multicenter Allergy Study. AB - Linkage of asthma and high total serum IgE levels to chromosome 12q15-q24.1 has been recently described. To evaluate this region further in regard to total IgE responsiveness, we genotyped 52 unrelated German children with persistently "high" total serum IgE (selected from a noninterventional prospective multicenter cohort study) and their parents. We carefully defined a most extreme IgE phenotype and analyzed it as a dichotomous trait. We tested for linkage between high total IgE concentrations and nine polymorphic microsatellite markers on chromosome 12q15-q24.1 using the transmission/disequilibrium test. Evidence for linkage and allelic association for high total IgE was observed for four markers in this region. This study demonstrates the value of using extreme phenotypes in genetic analysis of a complex quantitative trait. PMID- 9403075 TI - Automated capillary electrophoresis in the genotyping of apolipoprotein E. PMID- 9403076 TI - Radiation hybrid mapping of the genes for tenascin-R (TNR), phosducin (PDC), laminin C1 (LAMC1), and TAX in 1q25-q32. PMID- 9403077 TI - A low-copy repeat in Xq26 represents a novel putatively prenylated protein gene (CXX1) and its pseudogenes (DXS9914, DXS9915, and DXS9916). PMID- 9403078 TI - Scapula form and biomechanics in gorillas. AB - Gorillas are generating renewed interest as mounting evidence from field and molecular studies strongly suggests the western lowland (Gorilla gorilla gorilla) and eastern mountain (Gorilla gorilla beringei) gorillas are considerably more distinct than has previously been accepted. Schultz (1927, 1930, 1934) was one of the earliest investigators to document morphological differences between the two groups, noting differences in pedal, limb and scapular morphology. These differences led Schultz to conclude that while lowland gorillas retained some features suited to an arboreal habitat, the mountain gorilla had evolved into a specialized terrestrial quadruped. In particular, he noted that mountain gorillas exhibited lower values for the scapular index, higher values for ratios of infraspinous fossa vs. scapula length and spine length vs. scapula length and variability in the extent of curvature of the vertebral border. However, Schultz' observations were based upon small sample sizes of mostly adult specimens. This study extends Schultz' preliminary work by assessing, with appreciably larger sample sizes, patterns of relative growth of the scapula in these two subspecies of Gorilla. Scapula measurements were obtained for ontogenetic series of G.g. gorilla (n = 366) and G. g. beringei (n = 43). Statistical analyses reveal mountain gorillas exhibit significantly (P < 0.05) greater spine lengths and scapula breadths and smaller scapula lengths than lowland gorillas of comparable superior border lengths. However, at comparable body weights, mountain gorillas exhibit significantly shorter spines and superior borders than lowland gorillas. These differences in scapula proportions are evaluated in the context of biomechanical predictions regarding scapula form and locomotion. PMID- 9403079 TI - Experimental determinations of carcass processing by Plio-Pleistocene hominids and carnivores at FLK 22 (Zinjanthropus). Olduvai Gorge, Tanzania. AB - Published and unpublished skeletal and surface mark data from the large, well preserved, bovid dominated FLK 22 (Zinjanthropus) archaeofauna are analyzed using data derived from four different experimental control samples. The control samples are realistic because they are based on natural history and paleoecological data collected from FLK 22, and other Olduvai Gorge assemblages; they are precise because independent experimental studies following the same methods have generated the same results; and they restore generality to the study of site formation because each one models a different hominid and/or carnivore scenario of site formation. Comparability between FLK 22 and the control samples is established by excluding specimens from the former which do not meet identification and reporting standards derived from the latter. As in two previous studies, a comprehensive analysis of tooth marks and tool marks on long bone specimens from FLK 22 indicates that they were processed in three stages. In stage one, carnivores defleshed long bones, as inferred from the high percentage of tooth marks on midshaft fragments. In stage two, hominids processed intact long bones for marrow, as inferred from percussion mark percentages. Cut marks suggest that long bones retained flesh, but the amount, as yet, cannot be determined using cut mark percentages. In stage three, carnivores processed long bone epiphyses for grease, as inferred from the under-representation of long bone epiphyses and the high percentage of tooth marks on near-epiphyses and surviving epiphyses. The lack of comprehensive skeletal and surface mark data on cranial, axial, compact, and other specimens currently limits the application of experimental results. However, the available data suggest that the condition and representation of these items in the FLK 22 assemblage are also consistent with a carnivore to hominid to carnivore sequence of site formation. The variety of elements present, and their extensive processing by hominids, indicates that FLK 22 functioned as a central place/refuge where hominids could transport a variety of carcass parts and process them in an unhurried fashion. The presence of numerous small and medium sized individuals also indicates that hominids could have passively scavenged carcasses from a number of different sources including lions, leopards, sabertooth cats, and mass drownings. PMID- 9403080 TI - The occipital torus and developmental age of Sangiran-3. AB - Since its discovery in 1938 Sangiran-3 has been considered a juvenile Pithecanthropus (Homo) erectus, and therefore, excluded from studies of adult H. erectus. Although morphological features align Sangiran-3 with H. erectus, its age designation rests on an unconvincing reconstruction of the occipital torus and lack of sutural fusion. Evaluation of the occipital shows the original reconstruction is faulty and that the current midline occipital torus is actually the right lateral torus. The new reconstruction of Sangiran-3 results in midline toral morphology and development that is comparable with that in Sangiran-2. Compared with juvenile and adult H. erectus and Homo sapiens Sangiran-3 has three fully developed layers of vault bone with localized hypertrophy of the outer table into a sagittal keel, bregmatic eminence, and occipital torus. Sangiran-3's absolute vault thickness is also within the range of adult H. erectus. In addition, the coronal suture is fully interdigitated and sagittal sutural complexity is consistent with adult H. erectus. Sangiran-3's parietal sagittal contours are indistinguishable from adult H. erectus, whereas sagittal vault contours of juvenile H. erectus are usually more rounded than adults. These features indicate that Sangiran-3 is best considered a young adult H. erectus and should be included in metric and non-metric analyses of this taxon. PMID- 9403081 TI - Fuente Nueva-3 (Orce, Granada, Spain) and the first human occupation of Europe. PMID- 9403082 TI - Academy helps budding colleagues through ethics program. PMID- 9403083 TI - Delayed allergy-like reactions to X-ray contrast media. Exploration of the problem. PMID- 9403084 TI - [What is new? Therapy with lipid lowering agents]. PMID- 9403086 TI - [Bronchial asthma. Fluticasone propionate--new perspectives in the management of a frequently occurring disease entity]. PMID- 9403085 TI - [Sleep apnea and nightly hypertension. Interaction of sleep and respiration]. PMID- 9403087 TI - [Management of atherosclerosis--new findings for the practice]. PMID- 9403088 TI - [The effectiveness of the neurotropic agents: benfotiamin combinations]. PMID- 9403089 TI - [The metabolic syndrome]. PMID- 9403090 TI - [45th session of the Association of Ophthalmologists of Northwestern Germany. Lubeck, 1-2 June 1996. 46th Session of the Association of Ophthalmologists of Northwestern Germany, Bremen, 21-22 June 1997]. PMID- 9403091 TI - [Multiple sclerosis: from immunopathogenesis to disease management]. PMID- 9403092 TI - [3 years of clinical experience with risperidone: new therapeutic standards in the management of schizophrenia]. PMID- 9403093 TI - [Thrombolysis: Reteplase as an innovative alternative]. PMID- 9403094 TI - Behavioral responses of Barbary macaques (Macaca sylvanus) to variations in environmental conditions in Algeria. AB - In this study, the behavioral responses of Barbary macaques to seasonal and interhabitat variations in resource availability were analyzed over an entire annual cycle. Two groups, one in an evergreen cedar-oak forest (Djurdjura) and the other in a deciduous oak forest (Akfadou), were observed. In this paper, references to data on resource availability published elsewhere are made. Time budget has been studied. Variations in foraging and moving time, in day-range lengths, and in time moving in trees have been considered to estimate the variations in foraging effort and thus energy expenditure. Great monthly variations in foraging effort and other activities were observed in both habitats. In early spring, when resource availabilities were maximal, foraging effort was low while monkeys maximized their feeding time (about 5 h/day). In June, during the peak of the birth season and the rearing period, monkeys minimized their feeding time to the benefit of social interactions (to 1.6-2.7 h/day), whatever the food availability, which was low in Akfadou and high in Djurdjura. In addition, foraging effort remained low in Djurdjura, while it increased in Akfadou. Thus, at the beginning of the dry summer period, monkeys in Akfadou were in a less favorable position than those in Djurdjura. At both sites, in periods of food shortage in summer or in winter, monkeys displayed two different strategies. In the former case, their foraging effort increased, while in the second one it remained relatively low. Whatever the foraging effort, monkeys did not reach the same amount of feeding time as in early spring. In the poorest site of Akfadou, foraging effort was globally greater than in the richest site of Djurdjura, especially for adults. At both sites, adult males spent more time feeding than juveniles and less time in social interactions. Results are discussed according to rearing period, temperatures, and day length constraints. The limits of adaptability to different habitats are considered in light of the demographic parameters. PMID- 9403095 TI - Seasonal variation in the feeding behavior and diet of Japanese macaques (Macaca fuscata yakui) in lowland forest of Yakushima. AB - The feeding behavior of the southern subspecies of Japanese macaque (Macaca fuscata yakui) was studied over a period of 18 months in warm temperate broad leaved forest on the island of Yakushima, Japan. Focal animal data were collected for the eight adults in the troop. Over a full annual cycle, 35.0% of foraging on identified foods was on leaves and shoots, 30.2% on fleshy fruit, 13.2% on seeds, and 5.5% on flowers. Invertebrates and other animal matter accounted for 10.3% of foraging and fungi for 4.6%. There was marked seasonal variation in the use of different food categories, and seeds, leaves, fleshy fruit, and animal matter were each predominant at different times of year. There was also evidence of annual cyclicity in patterns of foraging on all major food types. The monkeys spent less time moving and ate a greater variety of foods when feeding on leaves than when feeding on fruit and seeds, or on insects. Time spent foraging was positively correlated with diversity of the diet, but there was no simple relationship between time spent foraging and the predominant food type. This suggests that a wide variety of foods takes longer to harvest and process, irrespective of the food type. The diet of the study troop was flexible and could not be assigned to a simple dietary category, such as frugivorous or folivorous. If these data are representative of the subspecies, the Yakushima macaque is much more of a dietary generalist than most primates for which there are adequate data. PMID- 9403096 TI - Transvaginal ultrasonographic (TVS) evaluation of baboon gestation from 37-62 days postconception. AB - Our objective was to determine the growth of the embryo and surrounding structures during baboon (Papio anubis) gestation using transvaginal sonography (TVS). To this end, we evaluated 19 timed-mated baboons using TVS between 37 and 62 days of gestation. After visualization of the gestational sac, amniotic sac, and yolk sac, the three largest diameters of each of these extra embryonic structures were measured using longitudinal and transverse views. Embryonic crown rump length (CRL) was also recorded. Embryonic heart rates were determined using the M-mode function of the ultrasound equipment. All 19 gestations developed without complications. No significant trend could be demonstrated for heart rate or yolk sac diameters over the 37-62 day gestational age period. Mean (SD) gestational age in days, heart rate, and yolk sac diameter, respectively, for the group were 48 (7.8) days (range: 37-61), 180 (15) beats per minute (range: 156 221) and 5 (0.1) mm (range: 3-8). Significant correlations (P < 0.0001) were determined between gestational age and CRL and gestational and amniotic sacs. We conclude that TVS allows a clear visualization of the embryo proper and all the cavities within the gestational sac of the baboon gestation. This study has determined the normal pattern of changes of these cavities from 37-62 days of gestation. Future applications of these findings may include sampling fluid from these cavities for biochemical, cytological, and metabolic studies. PMID- 9403098 TI - Propagation of handclasp grooming among captive chimpanzees. AB - A grooming posture previously reported for two wild chimpanzee (Pan troglodytes) communities developed spontaneously in a captive group of the same species. This offered a unique opportunity to follow the propagation of a new social custom. The posture consists of two partners grasping hands--either both right hands or both left hands--and raising the arms in an A-frame above their heads while mutually grooming with their free hands. The propagation of this pattern was followed over a 5 year period. In the beginning, handclasps were always initiated by the same adult female. This female initiated the posture mainly with her adult female kin. In subsequent years, these relatives became frequent participants in the posture with each other as well as with nonrelatives. Over the years the posture increased in frequency and duration and spread to the majority of adults and also to a few adolescents and older juveniles. The pattern persisted after removal of the apparent originator. PMID- 9403097 TI - Reproductive events of wild cotton-top tamarins (Saguinus oedipus) in Colombia. AB - Reproductive patterns of wild cotton-top tamarin (Saguinus oedipus) females located in La Reserva Forestal Protectora Serrania de Coraza-Montes de Maria in Coloso, Colombia, were examined using long-term behavioral observations and fecal steroid analysis. Using an enzyme immunoassay, we analyzed fecal samples for E1C and PdG. Comparisons of reproductive cycles of a reproductively active female and her daughters were made. An inhibition of ovarian cycles has been observed in daughters living in their families. However, daughters also exhibited normal ovarian cycling that subsequently resulted in pregnancy. Factors influencing the fertility are discussed as they relate to the reproductive strategies of wild cotton-top tamarin females. PMID- 9403099 TI - Ovarian function of pygmy marmoset daughters (Cebuella pygmaea) in intact and motherless families. AB - Reproductive inhibition of subordinate Callitrichid group members has been shown to vary with genus; whereas female Leontopithecus subordinates have normal ovarian cycles and occasionally breed within groups, subordinate Saguinus females almost never do so, with Callithrix species showing intermediate levels of reproductive inhibition. No information has been available on patterns of reproduction or inhibition in subordinate females in the genus Cebuella. We assessed fertility in Cebuella pygmaea daughters to allow comparison with the remaining Callitrichid genera. Specifically, the project had two goals: 1) to determine if there was evidence of reproductive inhibition of daughters living in family groups, and 2) to compare the ovarian function of daughters living in intact family groups with that of daughters residing in motherless families (i.e., without the breeding female). We collected daily urine samples for 6-8 weeks from eight pygmy marmoset daughters living in intact family groups or motherless families. Determination of ovulatory cycling or reproductive quiescence depended on the hormonal profiles generated from urinary luteinizing hormone and pregnanediol-3-glucuronide concentrations. All females in the motherless condition (aged 13-30 months) (n = 4) ovulated during the study. In contrast, only one of four daughters (aged 13-20 months) residing in intact families was found to be cycling. Data from motherless groups indicate ovarian cycling may begin between 15 and 17 months of age. Reproductive inhibition occurs in pygmy marmosets, although in a pattern more similar to Callithrix than to other Callitrichid genera. PMID- 9403100 TI - Hypervariable microsatellite loci in the Japanese macaque (Macaca fuscata) conserved in related species. AB - We describe seven polymorphic microsatellites isolated from a Japanese macaque (Macaca fuscata) genomic library selected for (GT)n content. The primer sets amplified from four to 11 different alleles in a sample of 14 Japanese macaques from nine different sites along the central and southern distribution of the species. These heterologous primers also detected variability in four other cercopithecine species. PMID- 9403101 TI - DNA amplification polymorphisms of Mucor piriformis. AB - Mucor piriformis can cause postharvest decay in various fruits and vegetables stored at low temperatures. Thirty isolates of this fungus, collected from infected fruit, were subjected to random amplified polymorphic DNA (RAPD) analysis. Seven different 10-bp primers were used to determine the type and extent of intraspecific genetic polymorphisms. Nineteen composite amplification types were identified, indicating a higher degree of variability than found in previous isoenzyme studies. Numerical analysis with the UPGMA technique revealed three clusters, which correlated with the mating competency of the isolates or their place of origin. These results demonstrate that RAPD analysis can identify isolates and subspecific populations of M. piriformis. PMID- 9403102 TI - Pyrimidine biosynthesis in Pseudomonas stutzeri ATCC 17588. AB - The de novo pyrimidine biosynthetic enzymes in the denitrifying bacterium Pseudomonas stutzeri ATCC 17588 were assayed and their activities were lower in glucose-grown cells than in succinate-grown cells. When P. stutzeri was grown in the presence of uracil, the de novo enzyme activities in succinate-grown cells were lowered while they remained largely unchanged in glucose-grown cells. A uracil auxotroph of P. stutzeri, deficient for aspartate transcarbamoylase activity, was isolated and its auxotrophic requirement was met by only uracil and cytosine. The inability of pyrimidine ribonucleosides to meet the auxotrophic requirement was related to the limited ability of P. stutzeri to transport uridine and cytidine. Pyrimidine limitation of the auxotroph elevated the de novo enzyme activities indicating that this pathway may be repressible by a uracil related compound in succinate-grown P. stutzeri cells. Regulation of pyrimidine synthesis in P. stutzeri was similar to that observed for other pseudomonads classified within rRNA homology group I. PMID- 9403103 TI - Inhibition of denitrification activity but not of mRNA induction in Paracoccus denitrificans by nitrite at a suboptimal pH. AB - The influence of pH on the denitrification activity of a continuous culture of Paracoccus denitrificans was studied in relation to the presence of nitrite. After a transition from aerobic to anaerobic conditions at the suboptimal pH of 6.8, P. denitrificans was not able to build up a functional denitrification pathway. Nitrite accumulated in the medium as the predominant denitrification product. Although the nitrite reductase gene was induced properly, the enzyme could not be detected at sufficient amounts in the culture. These observations was somehow inhibited, or once synthesized nitrite reductase was inactivated, possibly by the high concentrations of nitrous acid (HNO2). Interestingly, when a P. denitrificans culture which was grown to steady-state under anaerobic conditions was then exposed to suboptimal pHs, cells exhibited a reduced overall denitrification activity, but neither nitrite nor any other denitrification intermediate accumulated. PMID- 9403104 TI - The production of gamma-linolenic acid by selected members of the Dikaryomycota grown on different carbon sources. AB - In this study, seven fungal strains, representing different phylogenetic groups within the Dikaryomycota, were tested for the presence of gamma-linolenic acid [18:3(omega 6)], when grown in synthetic liquid media devoid of fatty acids, on a series of 40 different carbon sources. The fungal strains represented the species Dipodascopsis uninucleata, Eurotium rubrum, Galactomyces geotrichum, Neurospora crassa, Saccharomyces cerevisiae, Spongipellis unicolor and Talaromyces flavus. Cultures were periodically harvested during growth and the fatty acids in the total lipids analysed as methyl esters, using gas chromatography and mass spectrometry. It was found that 18:3(omega 6) is present in E. rubrum CBS 350.65, S. unicolor CBS 117.16 and in T. flavus CBS 310.38NT, when these strains were grown on certain carbon sources. No correlation between the growth phase of the organism and the presence of 18:3(omega 6) could be detected. In order to confirm the production of 18:3(omega 6), the lipid metabolism of two unrelated dikaryomycotan fungi (S. unicolor CBS 117.16 and E. rubrum CBS 350.65) grown on two different carbon sources each, was examined. Cultures of E. rubrum CBS 350.65 were grown on glucose and sorbose and cultures of S. unicolor CBS 117.16 on glucose and sucrose in synthetic liquid media with a C:N ratio of 50:1 (w/w). The total lipids of these cultures were fractionated and the fatty acids in the fractions analysed as methyl esters, using gas chromatography and mass spectrometry. The lipid metabolism of both E. rubrum CBS 350.65 and S. unicolor CBS 117.16 differed on the two carbon sources used. The ab initio production of 18:3(omega 6) by E. rubrum CBS 350.65 in synthetic liquid media was confirmed. In contrast, the ab initio production of 18:3(omega 6) by S. unicolor CBS 117.16 in synthetic liquid media could not be confirmed. PMID- 9403105 TI - Assimilation of volatiles from ripe apples by Sporidiobolus salmonicolor and Tilletiopsis washingtonensis. AB - Sporidiobolus salmonicolor ATCC 623 and Tilletiopsis washingtonensis NRRL Y-2555 grew on carbon resources provided as volatiles by ripe 'Golden Delicious' apples. This ability was not correlated with the reported natural habitats of the 21 species (26 strains) tested. Ethylene, the major volatile produced, was not utilized but butyl acetate, hexyl acetate and hexyl-2-methyl-butanoate (identified by GC-MS) were. These yeasts also assimilated ethanol, butanol, hexanol (Tilletiopsis excepted), acetate, propionate, butyrate and ethyl acetate at appropriately low concentrations. Ethanol and acetate aside, this is the first report of such assimilations by any yeast. PMID- 9403106 TI - Evolutionary advances in the higher fungi. AB - In this report the phrase 'evolutionary advances' is used in three ways: 1. to describe monophyletic changes perceived within a lineage; 2. to describe evolutionary sequences that appear to have become parallel/convergent; 3. to describe major transitions inferred between primary taxa. In monophyletic evolution the changes occur within a specific lineage that arises from a common ancestor, e.g., modern Man, horses, rusts. In parallel/convergent evolution different lineages respond similarly over time to environmental challenges and opportunities and come to acquire a great deal of comparability (e.g., Webster, 1987). Such lineages may be designated as separate taxa, e.g., similarities in marsupial and placental carnivores, or, if the polyphyleticism is cryptic, as a collective taxon, e.g., Aves, the class of birds, the obsolete Amentiferae for catkin bearing plants, the Gasteromycetes, and lichens. In major transitions there are significant paradigm shifts in which evolutionary changes from one predominant life style pattern to another are accompanied by increases in complexity (see Smith & Szathary, 1995), e.g., symbiosis, the water to land transition, the changes between the phyla of land fungi. Three particular terms are used in evaluating evolutionary relationships (Moore, 1996a): homology, paramology, and analogy. Homology, from Darwin's theory of common descent, is the phenomenon of having a common historical origin but not necessarily the same final structure or function (e.g., vertebrate forelimbs). Paramology (Moore, 1971) applies to inferred relationships in evolutionary schemes based on contemporary forms that lack fossil antecedents, e.g., the various phylogenetic interpretations of prokaryotes, algae, and fungi; Boekhout et al. (1993) have evaluated the taxonomic resolution of a variety of morphologic, biochemical, physiological, and molecular characters (Table 1). Analogy is generally applied to similar forms that are unrelated, e.g., insect/vertebrate wings; prokaryote/eukaryote flagella. It should also be borne in mind that, in a given taxon, biotrophism (Coffey, 1975) is an advanced character (Health, 1987) over, respectively, weaker parasitism, symbiotism, commensalism, and freeliving and that seemingly simple or less differentiated forms can be, and more than likely than not are, reduced, polyphyletic, and specialized rather than ancient and rudimentary, e.g., yeasts (Hoog et al., 1988; Kurtzman & Fell, 1996; Moore, 1988b; 1996a). PMID- 9403107 TI - MauE and MauD proteins are essential in methylamine metabolism of Paracoccus denitrificans. AB - Synthesis of enzymes involved in methylamine oxidation via methylamine dehydrogenase (MADH) is encoded by genes present in the mau cluster. Here we describe the sequence of the mauE and mauD genes from Paracoccus denitrificans as well as some properties of mauE and mauD mutants of this organism. The amino acid sequences derived from the mauE and mauD genes showed high similarity with their counterparts in related methylotrophs. Secondary structure analyses of the amino acid sequences predicted that MauE is a membrane protein with five transmembrane spanning helices and that MauD is a soluble protein with an N-terminal hydrophobic tail. Sequence comparison of MauD proteins from different organisms showed that these proteins have a conserved motif, Cys-Pro-Xaa-Cys, which is similar to a conserved motif found in periplasmic proteins that are involved in the biosynthesis of bacterial periplasmic enzymes containing haem c and/or disulphide bonds. The mauE and mauD mutant strains were unable to grow on methylamine but they grew well on other C1-compounds. These mutants grown under MADH-inducing conditions contained normal levels of the natural electron acceptor amicyanin, but undetectable levels of the beta-subunit and low levels of the alpha-subunit of MADH. It is proposed, therefore, that MauE and MauD are specifically involved in the processing, transport, and/or maturation of the beta subunit and that the absence of each of these proteins leads to production of a non-functional beta-subunit which becomes rapidly degraded. PMID- 9403109 TI - Growth, maintenance and fermentation pattern of the salt-tolerant lactic acid bacterium Tetragenococcus halophila in anaerobic glucose limited retention cultures. AB - The homofermentative lactic acid bacterium Tetragenococcus halophila showed mixed acid fermentation at low growth-rates under glucose limiting conditions and in the presence of 10% NaCl. Maximum growth yields in fermentors with cell retention were not affected by pH, but maintenance requirement was at pH 5.2 four times higher than at pH 7.0. Despite the high salt-concentration of the medium, maintenance requirements were low compared to other lactic acid bacteria. The possible causes of the observed differences in maintenance requirements are discussed. PMID- 9403108 TI - The incorporation of mannoproteins in the cell wall of S. cerevisiae and filamentous Ascomycetes. AB - In yeast, glucanase extractable cell wall proteins are anchored to the plasma membrane at an intermediate stage in their biogenesis via a glycosylphosphatidylinositol (GPI) moiety before they become anchored to the wall glucan via a beta 1,6-glucan linkage. The mechanism of the membrane processing step of cell wall proteins is not known. Here, we report that Ascomycete filamentous fungi involved in food spoilage such as Aspergillus, Paecilomyces and Penicillium, also contain GPI membrane-anchored proteins some of which are processed by an endogenous phospholipase C activity. Furthermore, similar to the situation in yeast, their cell walls contain mannoproteins which are linked to the glucan backbone through a beta 1,6-glucan linkage. Interestingly, one mould which contains a significant amount of non covalently linked beta 1,6 glucosylated cell wall proteins, is much more sensitive towards beta 1,3 glucanases and membrane perturbing peptides than the others. PMID- 9403110 TI - Staphylococcal serine proteinase increases intracellular free calcium concentration in human polymorphonuclear leukocytes. AB - Staphylococcal serine proteinase (SSP) can influence various functions of human polymorphonuclear leukocytes (PMNL) including chemotaxis and phagocytosis. Since the rise in intracellular free calcium concentration is an important step in signal transduction leading to phagocyte activation, we tested the ability of SSP to increase the intracellular free calcium concentration in human PMNL using the fluorescent calcium indicator Fura-2AM. PMNL isolated from healthy donors responded to SSP in the concentration range of 10 to 100 micrograms/ml. The highest Ca2+ rise (104 +/- 47 nM) was observed for 10 micrograms/ml SSP. It was mainly dependent (81 +/- 11%) on extracellular calcium influx, however, SSP mobilized 68 +/- 7% of Ca2+ from intracellular calcium stores. Boiling of SSP or preincubation with phenylmethylsulphonylfluoride (an serine proteinase inhibitor) did not change its ability to increase intracellular free calcium concentration in PMNL. It suggests that active center of SSP is not responsible for Ca2+ mobilization. Finally, PMNL responded to each of three consecutive stimulations with SSP independently of the presence of high or low extracellular Ca2+ concentration. This may be an additional mechanism responsible for activation of human PMNL and degradation of alveolar walls during the staphylococcal infection in the lower airways. PMID- 9403111 TI - Molecular evolution and optimization. AB - Microbial populations (and life) not only evolve, they optimize. The transition from a random, unorganized, lifeless Earth to the present situation, where the Earth is virtually covered with nucleic acids and diverse and complex species, required numerous molecular changes and the integration of metabolic pathways over billions of years. Primitive prokaryotic life was dependent on and constrained by the physical-chemical conditions on the Earth, while slowly reshaping conditions present. In this review, molecular evolution and molecular optimization are examined with an emphasis on the order in which evolutionary events occurred. PMID- 9403112 TI - Soil mycoflora from the Dead Sea Oases of Ein Gedi and Einot Zuqim (Israel). AB - Samples were taken from the top 10 cm of soils from 24 points in the Ein Gedi area. Among 329 isolates, 142 species were identified: 11 genera of ascomycetes, one genus of coelomycetes, 28 genera of hyphomycetes, 7 genera of zygomycetes and one yeast, in addition to some unidentified basidiomycetes. The hyphomycetes were represented by 17 dematiaceous, 9 mucedinaceous and two tuberculariaceous. Melanconiaceous and stilbellaceous genera were not found. Two new varieties of Microascus recently described were reisolated. No strict thermophiles or halophiles were obtained. There is apparently no very characteristic or specific fungal flora of the Dead Sea Oases although it was different from that found in the desert soil surrounding this area. PMID- 9403113 TI - Investigation of calcium-binding sites on the surfaces of selected gram-positive oral organisms. AB - Dental plaque is rich in anionic groups with a high calcium-binding capacity which may affect mineral dynamics at the tooth surface. The two major calcium binding sites on Gram-positive cell surfaces are carboxylate groups (in proteins and peptidoglycan cross-links) and phosphate groups (in lipoteichoic and teichoic acid). Equilibrium dialysis was used to measure calcium-binding capacities of whole cells and purified cell-wall material (CWM) from Streptococcus mutans R9, Strep. oralis EF186, Strep. gordonii NCTC 7865, Strep. downei NCTC 11391, Actinomyces naeslundii WVU627 and Lactobacillus casei AC413. This material was stripped of phosphate (PS-CWM) and treated to mask carboxylate groups (CM-CWM). Whole-cell calcium-binding capacities ranged from 240 (Strep. downei) to 50 (L. casei) mu mol/g (dry wt). Differences in CWM, PS-CWM and CM-CWM calcium-binding capacities demonstrated the greater importance of phosphate in comparison with carboxylate groups in cell calcium binding. These data indicate that, in streptococci, calcium binding is predominantly phosphate group-based, especially in the teichoic acid-containing Strep. oralis. In the other species tested, calcium binding is predominantly carboxylate group-based. PMID- 9403114 TI - Caries prevalence in the permanent dentition of a mediaeval population from the south-west of Scotland. AB - The prevalence, distribution and location of dental caries were studied in the permanent dentition of skeletons from a large mediaeval cemetery, where successive phases of use could be distinguished. The main phases dated from 1240 AD to 1440 AD. During this 200-year period, caries prevalence showed a statistically significant linear trend to increase. There was an increase in caries prevalence with increasing age from age band 20-25 through 26-35 to 36-45, and this trend was statistically significant in all phase groups but one. The teeth attacked by caries were chiefly molars, followed by premolars, with a low rate of attack in incisors and canines. The differences in caries prevalence between these major tooth classes were significant. Juveniles and adults presented different patterns of carious attack on tooth surfaces, occlusal surfaces being most frequently affected in juveniles, and approximal surfaces in adults. The overall caries prevalence in the mediaeval population of Whithorn was 6.4% of the teeth present, a figure similar to those published for other Scottish mediaeval groups, but lower than the caries prevalence in an English mediaeval group. PMID- 9403115 TI - Nitric oxide synthase activities in mammalian parotid and submandibular salivary glands. AB - Nitric oxide is important as a physiological messenger molecule in various organs and cells. It is synthesized from the amino acid L-arginine by nitric oxide synthase. Here, the specific activities of nitric oxide synthase in the cytosolic fractions of rabbit, bovine, mice, rat, and guinea-pig parotid and submandibular glands were compared. Marked specific activities were detected in the rabbit and bovine parotid and submandibular glands and in the parotid of mice. The activity in rabbit parotid was highest and was similar to that in rabbit brain. The significant activities in the salivary glands were completely blocked in the absence of Ca2+ or the presence of a calmodulin inhibitor. These findings suggest that the rabbit parotid glands are useful for studying the regulation of nitric oxide generation by Ca2+/calmodulin-dependent nitric oxide synthase in salivary glands. PMID- 9403116 TI - Spatiotemporal expression of the homeobox gene S8 during mouse tooth development. AB - The murine S8 gene encodes a nuclear homeodomain containing transcription factor that is expressed at sites of epithelial-mesenchymal interactions, including those in cranofacial tissues. The spatiotemporal expression of S8 mRNA was examined in tooth primordia by in situ hybridization. S8 transcripts were found in all stages of tooth development in 13- to 16.5-day-old mouse embryos (E13 E16.5), covering the early bud stage up to the late bell stage. S8 mRNA was found exclusively in the ectomesenchyme and its derivatives that originate from the neural crest: future pulp cells, odontoblast precursors and dental follicle cells. Expression was highest at the late cap and early bud stages and declined at the mid-bell stage, in both first molar and incisor primordia. In E13 jaw explants grown in organ culture for 48 h, S8 mRNA was still present in first and second molar primordia after culture. At E15.5, S8 mRNA was also transiently present in the surrounding osteogenic tissue. It is concluded that the distribution pattern of S8 mRNA during tooth development indicates a role for the gene in defining the identity of dental papilla and follicle cells. It is speculated that the time-restricted expression of S8 in tooth primordia involves establishing the definitive form of the tooth organ. PMID- 9403117 TI - Comparison of calcium mobilization in response to noradrenaline and acetylcholine in submandibular cells of newborn and adult rats. AB - The response of mature and immature rat submandibular cells to alpha-receptor stimulation was compared in terms of the generation of inositol triphosphate (IP3) and Ca2+ mobilization, and of how the calcium mobilization response affects acetycholine (ACh)-induced Ca2+ mobilization. In mature cells, noradrenaline (NA) caused much smaller IP3 and Ca2+ responses than ACh. However, the Ca2+ release induced by NA was enough to partially discharge an agonist-sensitive store and to reduce Ca2+ release by a subsequent ACh stimulus. Exposure to NA also caused an influx of Ca2+ in the mature cells, which was largely associated with Ca2+ entry induced by store depletion (i.e. capacitative entry). In the immature submandibular cells of newborn rats, NA caused essentially no IP3 response and a small Ca2+ release, which only partially affected the Ca2+ released by a subsequent exposure to ACh. In contrast to adult cells, immature cells did not show an increased Ca2+ influx after exposure to NA. However, prestimulation with this agonist potentiated the Ca2+ influx activated by ACh in the cells of newborn rats, but not in cells of adult rats. As both mature and immature submandibular cells have a well-developed phosphoinositide turnover response to ACh, the findings in mature cells suggest a less efficient coupling between alpha receptors and phospholipase C, while those in immature cells suggest that this coupling is even less functional in the early stages of postnatal development. In permeabilized and 45Ca(2+)-loaded mature cells, cyclic ADP-ribose (cADPR) released 13.4% of loaded 45Ca2+ and this release was significantly reduced by pre exposure to IP3. Similarly, pre-exposure to cADPR also reduced the IP3-induced 45Ca2+ release. It is concluded that: (1) stimulation with NA induces a smaller Ca2+ release in mature and immature submandibular cells than ACh; (2) the mediator for this small Ca2+ mobilization may be cADPR; and (3) NA stimulates capacitative Ca2+ entry in mature cells, but not in immature cells, and it also activates a Ca2+ entry pathway distinct from the one induced by store depletion, particularly in immature cells. PMID- 9403118 TI - The influence of dental metal alloys on cell proliferation and fibronectin arrangement in human fibroblast cultures. AB - The biocompatibility of six single-phase dental metal alloys was studied by determining cell proliferation rates correlated to the arrangement of fibronectin (FN) in fibroblast cultures. Immunocytochemical methods were used to detect cell proliferation by 5-bromodeoxyuridine (BrdU) incorporation, and FN organization [i.e. diffuse in the extracellular matrix and organized in fibrils or in focal adhesions (FA)] in human fibroblast cultures. Cell proliferation rates were related to FN arrangement and in particular a higher percentage of cells in the S phase was related to a predominance of FA. The greatest difference in behaviour compared to that of the controls was detected after 120 and 168 hr: at these times, as well as at previous ones, the alloy with the highest Au content seemed the most biocompatible among those tested, as it behaved in a very similar way to the controls. In contrast, fibroblasts exposed to the other five alloys showed different behaviours from the controls. It is assumed that a correlation exists between FN organization and the percentage of BrdU-positive cells, and that these features vary in the presence of different alloys. The observation of FN arrangement together with cell proliferation rates could be another useful tool in determining the biocompatibility of dental metal alloys. PMID- 9403119 TI - Phenotypic characterization of mononuclear inflammatory cells in salivary glands of bio-breeding rats. AB - The purpose of this study was to assess whether mononuclear cell abnormalities exist in salivary glands from autoimmune Bio-Breeding (BB) rats. Frozen sections of gland tissues were prepared from five diabetes-resistant BB rats (BB-DR), from five BB rats with diabetes (BB-DP) and from five Wistar rats. A panel of six monoclonal antibodies was used to identify membrane antigens associated primarily with monocytes (ED1), mature tissue macrophages (ED2), lymphoid macrophages (ED3), MHC class II (Ia) antigen (OX6), CD5+ T lymphocytes (OX19), and rat B lymphocytes (OX33). Normal submandibular, sublingual and parotid glands contained few ED1-positive cells, usually two or fewer per field. Tissue macrophages identified by clone ED2 comprised a major mononuclear cell subset in both Wistar and BB rats. However, the number of ED2-positive mononuclear cells was significantly depressed in the submandibular and parotid glands from BB-DR and BB DP animals, being present in quantities 25-50% of those observed in glands from normal Wistar rats (p < 0.001). In contrast, 25- to 30-fold greater numbers of ED3-positive macrophages were observed in submandibular glands from BB rats (p < 0.001). MHC class II (Ia) antigen expression also was 4- to 6-fold greater in BB rat submandibular glands, compared to Wistar rats (p < 0.001). CD5+ T-lymphocytes were rare or entirely absent in BB sublingual glands (0 to 1 cell per 0.87 mm2 field), compared to 47 cells per field from Wistar sublingual glands. No B lymphocytes were identified with antibody OX33 in any of the rat strains. These findings indicate that BB rat salivary glands differ significantly from Wistar salivary glands. In BB rats there is a rich population of ED3-positive macrophages and T lymphocytes in submandibular gland, low quantities of T lymphocytes in sublingual gland, and fewer ED2-positive macrophages in all three major salivary glands. These differences in mononuclear cell subpopulations may also influence salivary gland function in mucosal immunity. PMID- 9403120 TI - Dissolution of powdered human enamel suspended in acid solutions at a high solid/solution ratio under a 5% CO2 atmosphere at 20 degrees C. AB - The aim was to examine the nature of enamel dissolution in aqueous suspensions with a high solid/solution ratio and in a CO2-rich atmosphere. Before experimentation, a water-saturated mixture of 95% N2-5% CO2 was passed through the acid solutions for 24 hr. Samples of 2 g of powdered enamel were suspended in 7 ml of either 5 or 10 mmol/l HClO4, with or without 2 parts/10(6) fluoride and kept gently agitated for 24 hr in the above atmosphere. The same enamel samples were repeatedly exposed to fresh acid for 26 runs. All experiments were duplicated. The aqueous phase was analysed after 20 min and 24 hr for calcium, phosphate, fluoride, chloride, sodium and magnesium. It was found that after 20 min the fluoride was invariably taken up in the enamel and the solution was supersaturated with respect to hydroxyapatite with pH ranging 6.7-5.6. During the following 24 hr pH increased further, the supersaturation remained unchanged and the concentrations of calcium and phosphate in solution decreased. In contrast, sodium, magnesium and chloride were released from enamel during the entire period. In the later runs, the supersaturation with respect to hydroxyapatite was only modest and the decrease of calcium and phosphate concentrations limited, as were the release of sodium, magnesium and chloride. It is concluded that despite a CO2-rich atmosphere, calcium, phosphate and carbonate were released from enamel and quickly established a supersaturation with respect to hydroxyapatite with a secondary reprecipitation of mineral. It indicates that within the dental caries lesion in vivo, lesion fluid cannot exist undersaturated with respect to enamel apatite. PMID- 9403121 TI - Growth hormone secretion in the elderly: ageing and the somatopause. AB - The syndrome associated with lack of growth hormone (GH) in adults can be reversed by treatment with recombinant human GH (rhGH) with apparently beneficial clinical effects. This syndrome is strikingly similar to the characteristics of normal older adults which are known as the somatopause. GH secretion and insulin like growth factor I levels are reduced in healthy older people and it has been suggested that the somatopause is an age-related GH deficiency state. This review describes the physiological control of GH secretion in adults and seeks an explanation for the age-related decline, considering the impact of other factors such as nutrition and mobility, and particularly whether exercise offers a physiological approach to changing both the GH decline and the somatopause. The benefits and side-effects of treatment with rhGH for normal older people or older patients facing catabolic stresses are reviewed together with alternative approaches to stimulate GH such as GH-releasing hormone and the new pharmaceutical GH secretagogues. PMID- 9403122 TI - Thyroid diseases in the elderly. AB - The ageing thyroid is associated with a number of morphological and functional changes, such as decreased serum T3 and mean thyroid-stimulating hormone concentrations, that are to some extent independent of intercurrent non-thyroidal illnesses. All thyroid diseases, including clinical and subclinical hypo- and hyperthyroidism, non-toxic nodular goitre and thyroid cancer, are encountered in the elderly, but their prevalence and clinical expression differ from those observed in younger patients. In the elderly, autoimmune hypothyroidism is particularly prevalent, hyperthyroidism is mainly characterized by cardiovascular symptoms and is frequently due to toxic nodular goitres, and differentiated thyroid carcinoma is more aggressive. The interpretation of thyroid function tests is difficult in old individuals, because of age-associated changes in thyroid function and frequent alterations secondary to non-thyroidal illnesses and/or drugs. Treatment of thyroid disease deserves special attention in old patients because of the increased risk of complications. PMID- 9403123 TI - Ageing and adrenal cortical function. AB - Ageing is associated with changes in the secretion of adrenal cortical steroids. In the elderly, cortisol secretion increases after stimulation, while the secretion of dehydroepiandrosterone (DHEA) decreases. Each of these hormones influences the age-related processes of energy metabolism, fat depot distribution, immune function and neurodegeneration. In animals the effects of adrenal steroids are dramatic and easily measured. In humans the effects are more subtle. This review summarizes these actions and emphasizes the differences of dosages used in various experimental designs. It is concluded that adrenal hormones may play a significant role in human ageing, but research is hindered because the molecular pathways of DHEA action are not known. PMID- 9403124 TI - Declining gonadal function in elderly men. AB - Ageing in men is accompanied by a progressive decline of gonadal function with, in particular, a decline of total and free testosterone (T) plasma levels resulting in a significant proportion of elderly men over age 60 years presenting with subnormal T levels compared with the levels in young adults. A great interindividual variation in T levels is observed in elderly men, a variability explained in part by physiological variables and differences in life style, while associated acute or chronic diseases may accentuate the age-related decline of T levels. The progressive decrease of plasma T levels has been shown to result from both primary testicular changes and altered neuroendocrine regulation of Leydig cell function. At present, little is known about the clinical relevance of the relative hypoandrogenism of elderly men and there is an urgent need for more longitudinal studies, which may clarify a possible role of decreased T levels in the modulation of the clinical consequences of ageing in men. In view of the lack of relevant controlled clinical trials having careful assessment of the risks and benefits of androgen replacement therapy in elderly men, this treatment should be reserved for selected patients with clinically and biochemically manifest hypogonadism, after careful screening for contraindications. PMID- 9403125 TI - Menopause and post-menopause. AB - From the endocrine point of view, menopause is considered a deficiency state and oestrogen therapy regarded as restoring the pre-menopausal endocrine milieu. Oestrogen therapy alleviates acute climacteric symptoms and also reduces the risk of cardiovascular disease, osteoporosis and Alzheimer's disease. Cardiovascular protection seems to be the major benefit of oestrogen replacement: it reduces morbidity and mortality from coronary heart disease by approximately 50%. The mechanisms are complex and not fully under-stood. In this review we discuss currently available data on the effects of hormone replacement therapy on serum lipids and lipoproteins, the vessel wall (endothelium dependent and endothelium independent), blood flow, cardiac function, blood pressure, haemostasis, insulin sensitivity and direct anti-atherosclerotic effect as possible mechanisms of cardioprotection. Oestrogen therapy reduces the rate of post-menopausal bone loss, increases bone mineral density (BMD) and decreases fracture rate. Recent evidence suggests that initiation of oestrogen therapy in older women produces larger increases in BMD which might provide a significant protective effect at the time when fracture is common. The incidence of Alzheimer's disease is reduced by 50% in post-menopausal women taking oestrogen replacement. Limited clinical trials of oestrogen treatment in women with this disease have documented beneficial effects on cognitive function. The results of epidemiological studies of the effects of oestrogens on breast cancer risk are conflicting but recent evidence suggests that the risk is increased in current users after 5 years of use and among older women. In contrast, increase in the risk of venous thromboembolism is most significant within the first 12 months of therapy, strongly suggesting the importance of individual susceptibility. PMID- 9403126 TI - Ageing and calcium metabolism. AB - Ageing alters the metabolism of calcium and vitamin D in a number of ways. Intake of calcium and vitamin D, exposure to sunlight, cutaneous production of vitamin D3, renal production of 1,25-dihydroxyvitamin D (1,25(OH)2D3), intestinal absorption of calcium and the ability to adapt to a low calcium diet may all be reduced in elderly subjects. As a consequence, secondary hyperparathyroidism often occurs with ageing and can contribute to accelerated bone loss. In fact, alterations in calcium and vitamin D metabolism may be widespread in the ageing population and play a central role in the pathogenesis of senile (age-related) osteoporosis. From a preventive point of view, recent intervention studies have indicated the need to optimize calcium intake and to maintain serum 25(OH)D3 levels within the normal range in elderly people. PMID- 9403127 TI - Fluid and electrolyte homeostasis in the elderly: physiological changes of ageing and clinical consequences. AB - Characteristic of the normal ageing process are changes in the renal, hormonal and thirst regulatory systems involved in the control of sodium and water balance. In the presence of disease or drug use, the ageing changes put the elderly person at increased risk of either sodium retention or loss and of water retention or loss. Clinically, these alterations in water and sodium balance are commonly expressed as either hyponatraemia or hypernatraemia with central nervous system dysfunction as the symptomatic expression. Thus, the impaired homeostasis of the many systems affecting fluid balance in the elderly is readily influenced by many of the disease states and medications which are often present in the elderly with resultant adverse clinical consequences. Awareness of these age associated circumstances can allow the physician to anticipate the impact of illnesses and drugs and to implement a rational approach to therapeutic intervention and management. PMID- 9403128 TI - Non-insulin-dependent diabetes mellitus in the elderly. AB - The prevalence of non-insulin-dependent diabetes mellitus dramatically increases with age. Older diabetic subjects have an increased frequency of complications from diabetes compared with their younger counterparts and higher morbidity and mortality rates compared with age-matched non-diabetic controls. Elderly patients with diabetes are generally treated following the same approach as in younger patients: dietary therapy first, followed by oral hypoglycaemic agents and ultimately insulin. However, several specificities should be pointed out. Changes associated with ageing may affect the pharmacokinetics and pharmacodynamics of both sulphonylureas (increasing the risk of severe hypoglycaemia) and biguanides (increasing the risk of lactic acidosis). The best insulin regimen in old age is not known, but a twice-daily injection of a pre-mixed insulin preparation is usually recommended. Goals of therapy must be realistic and not cause disabling side-effects. The general practitioner plays a crucial role in the care of elderly diabetic patients, but access to a multidisciplinary specialized team may be necessary. PMID- 9403129 TI - Pituitary tumours in the elderly. AB - Pituitary tumours in the elderly are a neglected topic in the literature of endocrinology and gerontology. Data from autopsy studies in the elderly show that the commonest tumours are microadenomas, which are either immunocytochemically negative or stain for prolactin. Prolactin-secreting pituitary tumours, however, appear rarely in clinical series of patients in this age group and non functioning adenomas are the commonest tumour seen, although pituitary metastasis and craniopharyngiomas also occur. Diagnosis can be difficult, but the commonest presentation is visual field loss. Hypopituitarism may be diagnosed late and incidental radiological diagnosis is not uncommon. There are few data which specifically answer the question, but there is a suggestion that the syndromes associated with hormonal hypersecretion may be milder in this age group. Conventional treatment with drugs, surgery and radiotherapy should be decided on an individual basis as these therapies appear to be well tolerated and beneficial in selected patients. PMID- 9403130 TI - Effect of dietary selenium on the response of stressed and unstressed chickens to Escherichia coli challenge and antigen. AB - Selenium was added to the feed of White Leghorn type chickens 1 day prior to challenge with either Escherichia coli or sheep erythrocyte antigen. The incidence of death or lesions was reduced from 86% to 21% at the optimal dose of selenium (0.4 mg/kg resulting in feed concentration of 0.45 mg/kg). After the chickens were stressed by chilling, selenium was ineffective against E. coli. Dietary additions of selenium between 0.1 and 0.8 mg/kg resulted in an antibody titer increase from 2.2 to 3.9 to the log2 against sheep erythrocytes (SRBC). Following chilling, antibody titer response was reduced from 4.9 to 2.4 to the log2. This titer reduction could be prevented with dietary additions of selenium between 0.1 and 1.2 mg/kg. The effects of a nitrofuran and selenium were additive against E. coli challenge infection. PMID- 9403131 TI - Intranigral iron infusion in the rat. Acute elevations in nigral lipid peroxidation and striatal dopaminergic markers with ensuing nigral degeneration. AB - Iron is known to induce lipid peroxidation and recent evidence indicates that both iron and lipid peroxidation are elevated in the substantia nigra in Parkinson's disease (PD). To test whether excess intranigral iron induces lipid peroxidation, we infused an iron citrate solution (0.63 nmol in 0.25 microL) into the rat substantia nigra and measured nigral thiobarbituric acid reactive products at 1-h, 1-d, 1-wk, and 1-mo postinfusion. In a separate group of iron infused animals, histologic analysis within the substantia nigra through 1-mo postinfusion was accomplished by thionine- and iron-staining, with concurrent assessment of striatal neurochemical markers. Concentrations of nigral thiobarbituric acid reactive products were significantly elevated at 1 h and 1 d in iron-infused animals compared to vehicle-infused and unoperated animals, with a return to control values by 1 wk. Similarly, striatal dopamine turnover was acutely elevated, suggesting damage to dopaminergic neurons, which was confirmed histologically. Although iron-staining within the iron diffusionary area was increased through the postinfusion month, there was an apparent progression of the cellular character of staining from predominantly neuronal to reactive glial and finally to oligodendroglial by 1 mo postinfusion. This progression of cellular iron-staining may indicate a shifting of infused iron to a more bound unreactive form, thus explaining only an acute elevation in lipid peroxidation through 1 d following intranigral iron infusion. The data indicate that damage to nigral neurons induced by iron infusion is transciently associated with a marker of oxidative damage and supports the possibility that iron-induced oxidative stress contributes to the pathogenesis of PD. PMID- 9403132 TI - Changes in certain hematological and physiological variables following single gallium arsenide exposure in rats. AB - Gallium arsenide (GaAs), a group III-VA intermetallic semiconductor, possesses superior electronic and optical properties and has a wide application in electronic industry. Exposure to GaAs in the semiconductor industries could be a possible occupational risk. The aim of the present study was to determine the dose-dependent effect of single oral exposure to GaAs (500, 1000, or 2000 mg/kg) on some biochemical variables in heme synthesis pathway and few selected physiological variables at d 1, 7, and 15 following administration. The results indicate that GaAs produced a significant effect on the activity of delta aminolevulinic acid dehydratase (ALAD) in blood and heart (particularly at d 7) following exposure to 2000 mg/kg, whereas urinary delta-aminolevulinic acid (ALA) excretion was elevated only at d 7. No marked influence of GaAs on blood hemoglobin, zinc protoporphyrin, and packed cell volume was noticed. Blood glutathione (GSH) was significantly reduced at d 7, but remained unchanged at two other time intervals. On the other hand, heart GSH contents remained uninfluenced on GaAs exposure. Most of the physiological variables, viz. blood pressure, heart and respiration rate, and twitch response, remained unchanged, except for some minor alterations observed at d 7 and 15 following exposure to GaAs at a dose of 2000 mg/kg. Blood gallium concentration was not detectable in normal animals and rats exposed to 500 mg/kg GaAs. Blood arsenic concentration was, however, detectable even at the a lower dose level and increased in a dose-dependent manner. All these changes showed a recovery pattern at d 21, indicating that the alterations are reversible. PMID- 9403133 TI - Trace element status of hemodialyzed patients. AB - The trace elements Ba, Bi, Cd, Co, Cs, Cu, Hg, La, Mn, Mo, Pb, Rb, Sb, Sn, Sr, Tl, and Zn were determined by inductively coupled plasma mass spectrometry in plasma samples of 68 hemodialysis patients. The same elements (with exception of La and Mn) were also determined in whole blood after mineralization with high purity nitric acid/hydrogen peroxide in a closed-pressurized microwave system. The accuracy and precision was checked by analyzing two Seronorm "whole blood" reference materials. All samples were contaminated with barium (heparinized tubes) and the plasma samples with tin (collection tubes). The concentrations for Bi, Hg, Pb, Rb, Sb, and Sr in whole blood were within the literature ranges for healthy adults. All of the concentrations for Co, and some of the concentrations for Cd, Cs, Tl, and Zn were higher than the high limits of the normal ranges. Approximately 14% of the Cu concentrations were lower than the low limit of the normal range. The Mo and Sn concentrations are difficult to evaluate, because the normal ranges appears to be unreliable. All concentrations for Cd, Co, Mo, Pb, Sn, and Sr and some of the concentrations for Cu (15%) and Mn (75%) in the plasma samples were higher than the high limits of the normal ranges. The concentrations for Rb tended to be lower than the normal range. To establish unequivocally the causes for elevated and reduced concentrations of trace elements in whole blood and plasma of dialysis patients, all fluids in the dialysis process must be investigated. PMID- 9403134 TI - PIGE-PIXE analysis of medicinal plants and vegetables of pharmacological importance. AB - PIGE and PIXE techniques were employed to the study of elemental constituents of some traditional medicinal plants generally used in curing many diseases and ailments in southwestern Nigeria. Analyses were also carried out on commonly edible vegetables of medicinal and pharmacological importance. PIGE measurements were carried out using 3.5-MeV collimated protons from the 7 mV CN Van-de-Graaff accelerator of INFN, LNL, Legnaro (Padova), Italy, whereas the PIXE measurements were carried out using 1.8 MeV from the 2.5 MV AN 2000 Van-de-Graaff accelerator of the same laboratory. The results show that many of the medicinal plants contain elements of cardinal importance in human metabolism. The results from the vegetables also show the presence of vital elements that are needed for growth and development. In addition, some of the toxic elements, which include As, Cd, Hg, and so forth, were not detected. However, some of the recipes contain trace amounts of Pb at very low concentrations. This calls for proper control of dose rates in some samples to prevent the attendant negative cumulative effects. PMID- 9403135 TI - Dietary selenium- and vitamin E-induced alterations in some rabbit tissues. AB - The present study was designed to investigate and compare the effects of dietary selenium (Se) and vitamin E on some physiological parameters and histological changes in liver, heart, and skin tissues, as well as the blood parameters and the related enzymes. Both sex young rabbits were fed with deficient (9.8 micrograms/kg diet), adequate (225 micrograms/kg diet), and rich (4.2 mg/kg diet) Se and vitamin E diets for 12-15 wk for this purpose. As the plasma Se levels and the erythrocyte glutathione (GSH) peroxidase activity decreased (79.8 +/- 9.4 ng/mL and 2.0 +/- 0.3 U/g Hb, respectively) in the deficient group, these values increased (100.4 +/- 2.7 ng/mL and 14.5 +/- 4.3 U/g Hb) in the rich group significantly with respect to the control group. The other antioxidant enzyme activities and the related element levels did not change significantly in either one of the experimental groups. Although the platelet counts of the two experimental groups were not different from the control values, the collagen and the adenosine diphosphate (ADP) stimulated platelet aggregation rate and intensity increased in the deficient group (p < 0.05) and decreased very significantly (p < 0.001) in the rich group. In both of the experimental groups, as the percentage values of the neutrophils decreased, the lymphocytes and the eosinophils increased significantly. According to the light microscopic investigations, the observed lesions of considerable intensity within the tissues that elicit cell degenerations were more pronounced in the animals fed with the rich diet than in those fed with the deficient diet. The deficiency as well as toxicity of Se and the deficiency of vitamin E caused several alterations in the physiological functions of the tissues, and these alterations were supported by the histological lesions within these tissues. PMID- 9403136 TI - Bioavailability of rumen bacterial selenium in mice using tissue uptake technique. AB - A tissue uptake experiment was conducted to determine the bioavailability of rumen bacterial Selenium (Se) in mice. The donor animal was wether fed a diet containing 0.2 mg Se/kg dietary dry matter (DM). Ruminal fluid was collected 2 h postprandially. Bacterial-rich precipitate was obtained by differential centrifugation of the ruminal fluids. This was later freeze-dried and mixed in the diet to be used in feeding the mice experiment. Thirty growing female mice with a body wt (mean +/- SD) of 21.4 +/- 0.74 g were housed in plastic cages (5 mice/cage) and allotted equally to three dietary treatments. Diet 1 and Diet 2 were formulated based on AIN-76, except that no Se supplementation in the form of selenite was made in the former. In Diet 3, rumen bacterial matter was 20% of the diet, which gave an equivalent of 0.1 mg Se/kg dietary DM. The other two diets, Diet 1 and Diet 2, had an Se content of 0.025 and 0.1 mg/kg dietary DM, respectively. A 7-d feeding commenced after 7 d of acclimatization of the semipurified diet. Results showed that those mice fed an Se- (selenite) supplemented diet (Diet 2) had higher (P < 0.05) tissue Se concentrations than those mice fed the other two diets. No statistical differences were observed on various tissue Se concentrations between Diet 1 and Diet 3, although the latter diet had higher values. Kidney and liver had the highest Se concentrations compared to the other tissues. This study concludes that bacterial Se collected from the rumen of wether is not fully available for absorption in the intestine of the mice. PMID- 9403137 TI - Topology of ligand binding sites on the nicotinic acetylcholine receptor. AB - The nicotinic acetylcholine receptor (AChR) presents two very well differentiated domains for ligand binding that account for different cholinergic properties. In the hydrophilic extracellular region of both alpha subunits there exist the binding sites for agonists such as the neurotransmitter acetylcholine (ACh) and for competitive antagonists such as d-tubocurarine. Agonists trigger the channel opening upon binding while competitive antagonists compete for the former ones and inhibit its pharmacological action. Identification of all residues involved in recognition and binding of agonist and competitive antagonists is a primary objective in order to understand which structural components are related to the physiological function of the AChR. The picture for the localisation of the agonist/competitive antagonist binding sites is now clearer in the light of newer and better experimental evidence. These sites are mainly located on both alpha subunits in a pocket approximately 30-35 A above the surface membrane. Since both alpha subunits are sequentially identical, the observed high and low affinity for agonists on the receptor is conditioned by the interaction of the alpha subunit with the delta or the gamma chain, respectively. This relationship is opposite for curare-related drugs. This molecular interaction takes place probably at the interface formed by the different subunits. The principal component for the agonist/competitive antagonist binding sites involves several aromatic residues, in addition to the cysteine pair at 192-193, in three loops-forming binding domains (loops A-C). Other residues such as the negatively changed aspartates and glutamates (loop D), Thr or Tyr (loop E), and Trp (loop F) from non-alpha subunits were also found to form the complementary component of the agonist/competitive antagonist binding sites. Neurotoxins such as alpha-, kappa bungarotoxin and several alpha-conotoxins seem to partially overlap with the agonist/competitive antagonist binding sites at multiple point of contacts. The alpha subunits also carry the binding site for certain acetylcholinesterase inhibitors such as eserine and for the neurotransmitter 5-hydroxytryptamine which activate the receptor without interacting with the classical agonist binding sites. The link between specific subunits by means of the binding of ACh molecules might play a pivotal role in the relative shift among receptor subunits. This conformational change would allow for the opening of the intrinsic receptor cation channel transducting the external chemical signal elicited by the agonist into membrane depolarisation. The ion flux activity can be inhibited by non-competitive inhibitors (NCIs). For this kind of drugs, a population of low affinity binding sites has been found at the lipid-protein interface of the AChR. In addition, several high-affinity binding sites have been found to be located at different rings on the M2 transmembrane domain, namely luminal binding sites. In this regard, the serine ring is the locus for exogenous NCIs such as chlorpromazine, triphenylmethylphosphonium, the local anaesthetic QX-222, phencyclidine, and trifluoromethyliodophenyldiazirine. Trifluoromethyliodophenyldiazirine also binds to the valine ring, which is the postulated site for cembranoids. Additionally, the local anaesthetic meproadifen binding site seems to be located at the outer or extracellular ring. Interestingly, the M2 domain is also the locus for endogenous NCIs such as the neuropeptide substance P and the neurotransmitter 5-hydroxytryptamine. In contrast with this fact, experimental evidence supports the hypothesis for the existence of other NCI high-affinity binding sites located not at the channel lumen but at non-luminal binding domains. (ABSTRACT TRUNCATED) PMID- 9403138 TI - A circuitry model of the expression of behavioral sensitization to amphetamine like psychostimulants. AB - Repeated exposure to psychostimulants such as cocaine and amphetamine produces behavioral sensitization, which is characterized by an augmented locomotor response to a subsequent psychostimulant challenge injection. Experimentation focused on the neural underpinnings of behavioral sensitization has progressed from a singular focus on dopamine transmission in the nucleus accumbens and striatum to the study of cellular and molecular mechanisms that occur throughout the neural circuitry in which the mesocorticolimbic dopamine projections are embedded. This research effort has yielded a conglomerate of data that has resisted simple interpretations, primarily because no single neuronal effect is likely to be responsible for the expression of behavioral sensitization. The present review examines the literature and critically evaluates the extent to which the neural consequences of repeated psychostimulant administration are associated with the expression of behavioral sensitization. The neural alterations found to contribute to the long-term expression of behavioral sensitization are centered in a collection of interconnected limbic nuclei, which are termed the 'motive' circuit. This neural circuit is used as a template to organize the relevant biochemical and molecular findings into a model of the expression of behavioral sensitization. PMID- 9403139 TI - Cerebral cortex pathology in aging and Alzheimer's disease: a quantitative survey of large hospital-based geriatric and psychiatric cohorts. AB - In order to explore the relationships between the involvement of specific neuronal populations and cognitive deterioration, and to compare the hierarchical patterns of cortical involvement in normal brain aging and Alzheimer's disease, over 1200 brains from elderly subjects without cognitive deficits, as well as from patients with age-associated memory impairment and Alzheimer's disease, were examined. Our results suggest that the neuropathological changes associated with normal brain aging and Alzheimer's disease affect select cortical circuits at different points in time. Extensive hippocampal alterations are correlated with age-associated memory impairment, whereas substantial neurofibrillary tangle formation in neocortical association areas of the temporal lobe is a prerequisite for the development of Alzheimer's disease. Despite several lines of evidence involving amyloid deposit in the pathogenesis of Alzheimer's disease and Down's syndrome, our observations indicate that there is no correlation between senile plaque densities and degree of dementia in both disorders. In contrast to younger elderly cases, in the ninth and tenth decades of life, there is a differential cortical involvement in that parietal and cingulate areas are early affected in the course of Alzheimer's disease, and neocortical senile plaques densities are strongly correlated with the severity of dementia. Moreover, Alzheimer's disease symptomatology is characterized in these very old patients by high neurofibrillary tangle densities in the anterior CA1 field, but not in the entorhinal cortex and inferior temporal cortex. These observations are discussed in the light of the hypothesis of global corticocortical disconnection and with respect to the notion of selective neuronal vulnerability in Alzheimer's disease. PMID- 9403140 TI - Low-threshold Na+ currents: a new family of receptor-operated inward currents in mammalian nerve cells. AB - In the mammalian nervous system, various neurotransmitters can modulate cell excitability by inducing slow membrane potential changes. In the last decade, inhibition of potassium currents has been characterized as the primary mechanism by which neurones can undergo sustained depolarization. More recently (1990s), a new class of inward currents, which are voltage-dependent and mainly carried by sodium ions, has been found to be activated by various neurotransmitter receptors in mammalian central and peripheral neurones. Because the channels involved pass depolarizing current, are open at more negative membrane potentials than the resting potential, and are voltage-gated and persistent, these currents are capable of producing regenerative and maintained depolarizations and play an important role in neuronal signalling. PMID- 9403141 TI - Submissive behaviour and psychopathology. AB - OBJECTIVES: A variety of behaviours have been identified as submissive (Buss & Craik, 1986). These are believed to be associated with vulnerability to psychopathology. This paper explores the construct and measurement of submissive behaviours and their association with psychopathology. DESIGN: Two self-report scales were designed to measure the frequencies of (a) typical submissive behaviours (SBS) and (b) passive/withdrawal and affiliative strategies focused on conflict de-escalation (CDS). The association of these scales with psychopathology was explored in a series of questionnaire studies. METHODS: Study 1 assessed the SBS using a student sample (N = 332) and a mixed clinical group (N = 136). Of these, 177 students and 66 patients also completed the SCL-90-R. In Studies 2 and 3, the CDS and its association with depressive symptoms were assessed using a student sample (N = 154) and a depressed patient group (N = 60). RESULTS: The SBS and CDS appeared reliable. There was a positive relationship between the SBS and the SCL-90-R, including interpersonal sensitivity and unexpressed hostility. The passive/withdrawal subscale of the CDS was associated with depressive symptoms. Evidence was obtained for sex differences with the affiliative subscale. CONCLUSIONS: Some forms of submissive behaviour, especially those associated with passive/withdrawal and inhibition, are associated with a wide range of psychological problems. PMID- 9403142 TI - Selective processing of concern-related information in depression. AB - OBJECTIVES: The major question examined in this paper is whether selective attentional and interpretative processing of emotional information occurs in depression, and if so, whether it depends on a close match between the material used and current concerns. DESIGN: Twenty-four depressed patients and the same number of matched controls were tested using two selective processing tasks (described below), and their performance related to self-reported sociotropic and autonomy-related concerns. METHODS: Colour-naming interference and interpretation of ambiguous situations were assessed using material judged relevant to each of the Sociotropy-autonomy Scales. RESULTS: Depression was associated with a general interference effect for all negative concern words, and more negative interpretations of ambiguous situations, while controls showed a converse bias in favour of all positive interpretations. There was no convincing evidence that this negative processing bias was proportional to the match between material and self-reported sociotropic or autonomous concerns. CONCLUSIONS: Depressed patients showed evidence of cognitive biases favouring all negative self-related information, on both attentional and interpretative tasks. We suggest that such effects in depression may occur only under conditions allowing the elaborative processing of negative material related to oneself. PMID- 9403143 TI - Attributions and accessibility of explanations for future events in anxiety and depression. AB - OBJECTIVES: The research examined qualitative and quantitative aspects of future thinking in mood-disturbed participants. DESIGN: A cross-sectional, mixed design compared three groups of participants on measures of future thinking using an adapted fluency paradigm. METHODS: Participants who were either anxious (N = 25), anxious and depressed (mixed; N = 25), or neither anxious nor depressed (control; N = 25) were presented with a range of future positive and negative events and asked to provide explanations as to why those events would (pro reasons) or would not (con reasons) happen to them. Number of reasons given of each type was measured. The reasons were further analysed in terms of a number of attributional dimensions. RESULTS: Mood-disturbed participants (anxious and mixed) provided more pro relative to con reasons for negative events and more con relative to pro reasons for positive events. Compared to the control group, mood-disturbed participants also provided more internal and more global reasons for why negative events would happen and for why positive events would not happen. CONCLUSION: Mood-disturbed participants differ from controls on qualitative as well as quantitative aspects of future thinking. PMID- 9403144 TI - Interpersonal responses to threats to status and interpersonal relatedness: effects of dependency and self-criticism. AB - OBJECTIVES: Previous research investigating the interpersonal environments of dependent and self-critical individuals has focused primarily on attachment issues, such as relationship satisfaction. DESIGN: In the present study, we examined how dependent and self-critical individuals respond to experimentally manipulated events that threaten or bolster self-worth and status. METHOD: Forty pairs of female college students were allowed to believe, first, that they outperformed a close friend or were outperformed by a close friend on 14 trials of a behaviour detection task and, second, that friends generally agreed or disagreed with them on a second 14 trials, in which participants informed friends who had the better response. RESULTS: Dependent women were more concerned with maintaining interpersonal relatedness, whereas self-critical women were more concerned with preserving self-worth and status. Dependent women adopted the responses of friends they outperformed, praised friends even when friends disagreed, and minimized disagreement with disagreeing friends. In contrast, self critical individuals contested threats to status and self-worth, withheld praise from friends who challenged them, and did not minimize disagreement with disagreeing friends. CONCLUSIONS: Results support the utility of an interactional framework in which depressive personality styles, such as dependency and self criticism, and situational events interact to regulate interpersonal behaviour. PMID- 9403145 TI - Automaticity of cognitive biases in addictive behaviours: further evidence with gamblers. AB - OBJECTIVES: The hypotheses that automatic, non-volitional, attentional and memory biases for addiction-related constructs exist is tested with compulsive gamblers. DESIGN: An independent groups design was employed. Processing of gambling, compared to neutral and drug-related information was examined in 15 gamblers recruited from new members of Gamblers Anonymous. Comparisons were made with the performance of their spouses (N = 15) to help distinguish addiction mechanisms from more non-specific emotional experiences with gambling, and an independent control group (N = 15), recruited from the staff and students of a university department. METHODS: A modified Stroop procedure was first employed. Automative cognitive interference was assessed relatively, by comparing colour-naming times on the gambling, drug and neutral Stroops. A subsequent word-stem completion task of implicit memory was then used to assess selective and automatic priming of the gambling constructs in memory. RESULTS: Only the gamblers showed selective and automatic interference for gambling-related constructs on the Stroop task. Spouses behaved like the control group on this task. An implicit memory bias for gambling-related words was statistically detected only in the gamblers compared to the control group, although the trend was similar in the comparison with spouses. Further evidence for the specificity of these effects was obtained in subgroup comparisons involving fruit-machine with racing gamblers. CONCLUSIONS: Results are generally consistent with an automaticity in the cognitive biases gamblers show for gambling-related information. Implications for cognitive understanding and treatments are highlighted. PMID- 9403147 TI - Social reasoning in individuals with persecutory delusions: the effects of additional information on attributions for the observed behaviour of others. AB - The extent to which individuals suffering from persecutory delusions, as compared to matched controls, were prepared to change their attributional judgments following additional information, was investigated. Participants were required to decide whether the actor or target individual had caused an action before and after information, which was either high or low in terms of distinctiveness, consistency and consensus. There were no differences between the three participant groups which all made decisions, and changed their decisions, in the directions predicted by attribution theory. PMID- 9403146 TI - Relatives' locus of control and expressed emotion in schizophrenia and related psychoses. AB - OBJECTIVES: Knowledge of what predicts relatives' expressed emotion (EE) may contribute to improved family work in schizophrenia. In the present study we examined locus of control (LOC) beliefs as determinants of EE components. DESIGN: This study is observational, prospective and partly cross-sectional, partly longitudinal (stability of LOC). METHODS: In a Norwegian sample of 47 recently hospitalized patients (schizophrenia or schizophreniform disorder) and 70 relatives, the relatives' EE was assessed by the Camberwell Family Interview and LOC by Levenson's Internality, Powerful Others and Chance scales. RESULTS: Confirmatory multiple regression analyses showed that Chance LOC was positively related to emotional overinvolvement (p < .005). Powerful Others LOC, especially 'wish to ingratiate' items, were positively linked to criticism and, among workers/lower grade employees only, to emotional overinvolvement. Internal LOC was not linked to any EE scale. CONCLUSIONS: This study indicates that LOC beliefs may be determinants of emotional overinvolvement and criticism, and should be taken into account in family work that aims at modifying relatives' EE. PMID- 9403148 TI - The effect of self-referent material on the reasoning of people with delusions. AB - People with delusions have been shown to have both generalized (Huq, Garety & Hemsley, 1988) and content-specific biases in reasoning (Bentall, 1994). Our concern here was whether the hastiness that has been found when people with delusions reason on relatively abstract tasks would be present on a more realistic task. A second concern was whether reasoning with salient or emotional material would increase the hastiness bias in people with delusions. Two versions of a probabilistic reasoning task were used to study the data gathering of people with delusions. The first version employed realistic but emotionally neutral material. People with delusions requested less evidence before making a decision than psychiatric and normal comparison groups. Therefore, the hastiness found previously with abstract materials was seen to generalize to a more realistic task. In the second version participants were required to reason with material that had an emotional content and may have been regarded as being personally meaningful. In this condition all groups reduced the amount of evidence requested before making a decision. PMID- 9403149 TI - A three- to six-year follow-up of former long-stay residents of mental handicap hospitals in Northern Ireland. AB - OBJECTIVES: Little is known about the first cohorts of long-stay hospital residents with learning disabilities who moved to the community. This study describes the pattern of residential reprovision for all former long-stay residents discharged from the three mental handicap hospitals in Northern Ireland between 1987 and 1990 (N = 283) as well as describing aspects of quality of life for a smaller sample of people. METHOD: The study employs a retrospective survey design and the method and findings are discussed within a quality of life framework. Information about destinational outcomes between 1987 and 1993 was collected for each former resident. Several instruments were also used to assess material, emotional and social well-being and development and activity for a 40 per cent sample of people (114/283) discharged from hospital during 1987-1990 and followed up in 1993. RESULTS: Approximately 70 per cent of residents were discharged to, and subsequently remained in, highly supported settings such as residential and nursing homes. Only 3 per cent were discharged to 'independent living' with their own families or foster families. Few of the sample had 'major' problems with daily living skills and serious behavioural problems were uncommon. Former patients were also more satisfied with their new homes and reported feeling happier, healthier and more independent since discharge. However, social networks were poor and there was no evidence to suggest that people were undertaking new or 'ordinary' daytime activities. CONCLUSION: Although the material needs of former hospital residents (many of whom may have been 'cream skimmed' from the long-stay population) appeared to be met and they were content with their new homes in the community, they had a limited choice of mainly private sector accommodation and few opportunities for personal and social development. PMID- 9403150 TI - WAIS-R short forms: assessing the statistical significance of subtest differences. AB - OBJECTIVES: To assist with analysis, in the individual case, of subtest profiles obtained from WAIS-R short-form administrations. Secondly, to offer a method of profile analysis for a full-length WAIS-R which is appropriate for use with age graded scaled scores. DESIGN: The study used a psychometric method for profile analysis (Silverstein, 1982a) which deals more effectively than alternative approaches with the problems raised by conducting multiple comparisons. METHODS: A formula to compute the standard error of the difference between a subtest and mean subtest scores was applied to nine short forms and the full-length WAIS-R. The data used were derived from the WAIS-R standardization sample. RESULTS: Tables for examining whether there are significant subtest differences in a clients WAIS-R profile are presented. CONCLUSIONS: An elegant method of analysing subtest profiles has been extended to permit its use with WAIS-R short forms. Guidance on the use of the tables is offered; the distinction between reliable and abnormal differences is highlighted. PMID- 9403151 TI - Assessing the reliability and abnormality of subtest differences on the Test of Everyday Attention. AB - OBJECTIVES: To assist clinicians with the analysis of an individual's profile of subtest strength and weaknesses on the Test of Everyday Attention (TEA). DESIGN: The study applied psychometric methods for the quantitive analysis of subtest profiles (Silverstein, 1982, 1984a, b). METHODS: Formulae to compute the standard error of the difference and standard deviation of the difference between a subtest and a client's mean subtest scores were applied to determine critical values for reliable and abnormal differences. The data used were derived from the TEA standardization sample (N = 154). RESULTS: Tables for examining whether an individual's TEA subtest profile contains reliable and abnormal subtest discrepancies are presented. CONCLUSIONS: Elegant methods of analysing a subtest profile were extended for use with the Test of Everyday Attention. In keeping with the rationale underlying the measurement of neuropsychological deficit (Lezak, 1995), these methods complement the existing TEA normative comparison standards by providing individual comparison standards for a client's performance. Guidance on the use of the tables is offered; the distinction between reliable and abnormal differences is highlighted. PMID- 9403152 TI - Short NART, CCRT and Spot-the-Word: comparisons in older and demented persons. AB - This study compares the efficacy of three measures of premorbid intelligence: the National Adult Reading Test (NART), the Cambridge Contextual Reading Test (CCRT), and the Spot-the-Word Test. The results in a population sample of elderly and demented participants show that test efficacy varies between groups. In a demented group and a normal group of average readers, the CCRT leads to a higher estimate of premorbid word reading ability than the NART. Spot-the-Word results in good performance by normal groups and participants with minimal dementia, but performance is grossly impaired in subjects with mild/moderate dementia. We conclude that each test may be appropriate for specific groups. PMID- 9403153 TI - An evaluation of the Cambridge Contextual Reading Test (CCRT) in Alzheimer's disease. AB - Thirty patients with a diagnosis of probable Alzheimer's disease were assessed using the traditional National Adult Reading Test (NART) and also by placing the NART stimulus words in context (meaningful sentences)--the Cambridge Contextual Reading Test (CCRT) condition. Placing the stimulus words in sentences acted to significantly reduce overall pronunciation error rates. This beneficial effect was most marked for more severely cognitively impaired patients. NART performance was significantly correlated with Mini-Mental State total score; however, CCRT performance was not. Placing the stimulus words in context acted to improve the performance of more cognitively impaired Alzheimer patients and thus provides a more valid estimate of premorbid ability compared with the standard presentation of NART stimulus words in isolation. PMID- 9403154 TI - The discriminant validity of the Eating Disorder Inventory--2. AB - The scores of 78 bulimia nervosa (BN) patients and 67 general psychiatric outpatients on the Revised Eating Disorder Inventory (EDI-2) were compared in a multivariate discriminant analysis. The bulimia scale was found to correctly classify 97 per cent of all cases. Of the EDI-2 scales thought to be not directly related to food and weight, only interoceptive awareness (IA) and asceticism (AS) showed discriminative validity. Three eating-related items were found to account for the discriminative value of the IA scale, and the new AS scale was found to be a discriminative extension of the EDI. PMID- 9403156 TI - Schizotypal traits and dimensions of religiosity. AB - In the present study the association between religiosity and schizotypal traits was investigated. Two hundred and one respondents completed self-report measures of schizotypal traits as well as measures of religiosity developed, on the basis of theoretical work on the nature of delusional thinking, to tap the dimensions of preoccupation, guidance, conviction, and emotional involvement. For women, no associations were found between scores on the religiosity and schizotypy scales. For men, however, higher scores on religious preoccupation were associated with higher scores on magical ideation. It would seem that religiosity is not clearly associated with schizotypal traits. PMID- 9403155 TI - Processing of facial expressions of emotion and alexithymia. AB - OBJECTIVE: The primary objective was to examine emotional responsiveness in alexithymia. DESIGN: A quasi-experimental design was followed with the alexithymia variable being manipulated by subject stratification based on Toronto Alexithymia Scale-20-H. METHOD: Alexithymics (N = 12) and non-alexithymics (N = 12) were asked to match, label and verbally describe photographs displaying facial emotions along with TAT card II. RESULTS: Alexithymics did not differ from non-alexithymics in emotional matching and labelling tasks but had significant difficulty in verbally describing emotional expressions as evident by less duration of utterance, greater response latency and increased linguistic-type speech disruptions. Speech disruptions did not produce a group difference for TAT card II. CONCLUSION: Such difficulty in alexithymics may be associated with their inability to use emotional words in the appropriate context. PMID- 9403157 TI - Video games and clinical practice: issues, uses and treatments. PMID- 9403158 TI - Psychology and risk assessment. PMID- 9403159 TI - Cellular electrophysiologic mechanisms of cardiac arrhythmias. AB - Cardiac arrhythmias are caused by alterations in the electrophysiologic properties of the cardiac cells, which affect the characteristics of the transmembrane potentials. The electrophysiologic properties that cause arrhythmias are automaticity, triggered activity, and reentrant excitation. Each of these mechanisms is described in terms of the characteristics of the transmembrane potentials and how these influence the appearance of the arrhythmia on the electrocardiogram. PMID- 9403161 TI - Electrophysiologic evaluation of supraventricular tachycardia. PMID- 9403160 TI - Role of the surface electrocardiogram in the diagnosis of patients with supraventricular tachycardia. AB - In this era of interventional electrophysiology, the accuracy of the electrocardiogram in diagnosis of supraventricular tachycardia could be improved by detailed endocardial mapping and confirmed by results of radiofrequency catheter ablation. This article describes the electrocardiographic characteristics for different types of supraventricular tachycardia: atrial fibrillation, atrial flutter, atrial tachycardia, atrioventricular reciprocating tachycardia using an accessory pathway, and atrioventricular node reentrant tachycardia. Several limitations, including the identification of P wave morphologies and polarities and separation between the terminal part of T wave and P wave during tachycardia, should be resolved before an accurate algorithm of the 12-lead surface electrocardiogram is developed for the diagnosis of supraventricular tachycardia. PMID- 9403162 TI - Current role of pharmacologic therapy for patients with paroxysmal supraventricular tachycardia. AB - Intravenous antiarrhythmic drugs will continue to have an important role in the acute management of SVT. Long-term antiarrhythmic drug therapy is often effective in preventing or reducing frequency and severity of arrhythmic episodes. The cost, adverse effects, and inconvenience of long-term drug therapy will result in the increasing use of curative ablation for most individuals with problematic SVT. PMID- 9403163 TI - Inappropriate sinus tachycardia. Diagnosis and treatment. AB - Inappropriate sinus tachycardia is characterized by consistently elevated heart rates and exaggerated responses to minimal physiologic activity. The syndrome of inappropriate sinus tachycardia is defined by the clinical presentation of palpitations and presyncope that commonly appear to be out of proportion to the severity of the tachycardia. The development of a potentially curative procedure for patients suffering from inappropriate sinus tachycardia has renewed interest in the treatment of such patients. Continued research directed at the pathophysiology of inappropriate sinus tachycardia and what the optimal end point for achieving adequate rate control by radiofrequency catheter ablation without the need for the implantation of a permanent pacemaker is required. The optimal treatment may be the use of drugs that specifically inhibit sinus node pacemaker current. PMID- 9403164 TI - Catheter ablation for patients with atrial tachycardia. AB - Although atrial tachycardias are relatively rare, their poor response to standard therapies, the suboptimal hemodynamic results of complete atrioventricular node ablation and pacer implantation, and their potential for serious hemodynamic effects make management difficult. Although their mechanisms are complex and divergent, catheter ablation has proven to be highly effective in management of atrial tachycardias. This article discusses arrhythmia mechanisms and therapeutic approaches by catheter ablation. PMID- 9403165 TI - Catheter ablation for atrioventricular nodal reentrant tachycardia. AB - The current status of catheter ablation techniques for the management of atrioventricular nodal reentry tachycardia is outlined in this article. Some pertinent aspects of the atrioventricular nodal anatomy and physiology are discussed, to the extent that they are essential for understanding of the mechanism of this arrhythmia and the technique of catheter ablation. PMID- 9403166 TI - Catheter ablation of accessory pathways. AB - Radiofrequency catheter ablation is a highly effective, curative treatment for arrhythmias related to accessory atrioventricular connections. Compared with medical therapy, ablation is more definitive, is more cost-effective, and is associated with a lower risk of proarrhythmia. This article updates the reader on the current indications, techniques, and innovations related to ablation of accessory pathways using radiofrequency energy. PMID- 9403167 TI - Mechanisms and medical management of patients with atrial flutter. AB - Type I atrial flutter is due to reentrant excitation, principally in the right atrium. The standard ECG remains the cornerstone for its clinical diagnosis. Acute treatment should be directed at control of the ventricular response rate and, if possible, restoration of sinus rhythm. Radiofrequency catheter ablation therapy provides the best hope of cure, although atrial fibrillation may subsequently occur after an ostensibly successful ablative procedure. Alternatively, antiarrhythmic drug therapy to suppress recurrent atrial flutter episodes may be useful, recognizing that occasional recurrences are common despite therapy. Radiofrequency ablation of the His bundle ablation with placement of an appropriate pacemaker system may be useful in selected patients. PMID- 9403168 TI - The laboratory evaluation and role of catheter ablation for patients with atrial flutter. AB - The anatomic substrate for atrial flutter has now been recognized, and improved methods for catheter ablation have been developed. Using mapping techniques such as entrainment mapping, recognizing the different types of flutter that can occur, and testing for conduction block with pacing after ablation, long-term cure of atrial flutter can be achieved in most patients with catheter ablation. Not only is catheter ablative cure of atrial flutter the treatment of choice for drug-refractory patients, but also may now be offered as an alternative to drug therapy. PMID- 9403169 TI - The medical management of atrial fibrillation. AB - Atrial fibrillation is an extremely common arrhythmia that is associated with significant sequelae. Certain aspects of therapy, such as anticoagulation, are studied in well-constructed randomized trials. Other therapy, such as the maintenance of sinus rhythm with antiarrhythmic agents, is supported by limited evidence. This article reviews the epidemiology and medical treatment of this arrhythmia, addressing anticoagulation, ventricular rate control, and restoration and maintenance of sinus rhythm. Randomized trials in progress that attempt to answer important questions in the management of atrial fibrillation are also discussed. PMID- 9403170 TI - Radiofrequency catheter ablation for atrial fibrillation. AB - Until recently, catheter-based radiofrequency ablation for atrial fibrillation was limited to palliative approaches of either atrioventricular node ablation or modification. It is now recognized that at least a proportion of patients with paroxysmal atrial fibrillation may be suitable for curative ablation of an underlying single arrhythmogenic focus. With the intense interest in this area, a catheter-based cure involving endocardial linear lesion creation for patients with chronic or paroxysmal atrial fibrillation may not be far in the future. PMID- 9403171 TI - The maze procedure for cure of atrial fibrillation. AB - Atrial fibrillation is the most common dysrhythmia encountered in clinical practice. A significant number of patients fail medical therapy because of inability to convert or control the rhythm pharmacologically, intolerance of the requisite medication, or persistent symptoms despite apparently satisfactory rate control. Based on experimental studies establishing the electrophysiologic basis of atrial fibrillation, a surgical procedure has been developed that is highly effective in restoring sinus rhythm without further requirement for medications. The evolution of this procedure, its current indications, and results are outlined. PMID- 9403172 TI - Thermodynamics of peroxynitrite and its CO2 adduct. AB - The equilibrium constant, K3, of aqueous homolysis of peroxynitrous acid into hydroxyl and nitrogen dioxide free radicals was estimated to be 5 x 10(-10) M. This value was derived from a thermodynamic cycle by use of the experimentally known delta fH degree(ONOO-,aq) = -10.8 kcal/mol and the enthalpy of ionic dissociation of ONOOH(aq), delta H degree 1 = 0 kcal/mol, as well as of the entropy of gaseous ONOOH, S degree(ONOOH,g) = 72 eu. Furthermore we assumed the entropy of hydration of ONOOH, delta S degree 2, to be -25 eu, a value closely bracketed by the hydration entropies of analogous substances. The rate constant of radical recombination of OH. with NO2. to yield ONOOH, k-3, was resimulated from experimental data and found to be ca. 5 x 10(9) M-1 s-1. Together with the estimated K3, this yields the homolysis rate constant k3 = 2.5 s-1. This value is close to 0.5 s-1, the rate constant of formation of a reactive intermediate during the isomerization of peroxynitrous acid to nitrate. Our thermodynamic estimate is therefore consistent with substantial amounts of OH. and NO2. free radicals being formed in this process. The thermodynamic implications for the carbon dioxide/peroxynitrite system are also discussed. PMID- 9403173 TI - Glutathione conjugation of bay- and fjord-region diol epoxides of polycyclic aromatic hydrocarbons by glutathione transferases M1-1 and P1-1. AB - Metabolism of polycyclic aromatic hydrocarbons in mammalian cells results in the formation of vicinal diol epoxides considered as ultimate carcinogens if the oxirane ring is located in a bay- or fjord-region of the parent compound. In the present study, individual stereoisomers of the bay-region diol epoxides of chrysene, dibenz[a,h]anthracene, and benzo[a]pyrene as well as of the fjord region diol epoxides of benzo[c]phenanthrene, benzo[c]chrysene, and benzo[g] chrysene have been incubated with GSH in the presence of human glutathione transferases GSTM1-1 (a mu-class enzyme) and GSTP1-1 (a pi-class enzyme). As previously shown with GSTA1-1 (an alpha-class enzyme) both M1-1 and P1-1 demonstrate considerable activity toward a number of the diol epoxides studied, although a great variation in catalytic efficiency and enantioselectivity was observed. With GSTM1-1, the bay-region diol epoxides, in particular the syn diastereomers were in most cases more efficiently conjugated with GSH than the fjord-region analogues. GSTM1-1 demonstrated an enantioselectivity ranging from no preference (50%) to high preference (> or = 90%) for conjugation of the enantiomers with R-configuration at the benzylic position of the oxirane ring. With GSTP1-1, the enzyme demonstrated appreciable activity toward both bay- and fjord-region diol epoxides and, in most cases, a preference for the anti diastereomers. In contrast to GSTM1-1 and as previously shown for GSTA1-1, GSTP1 1 showed an exclusive preference for conjugation of the enantiomers with R configuration at the benzylic oxirane carbon. With both GSTM1-1 and GSTP1-1, the chemically most reactive diol epoxide, the (+)-syn-enantiomer of trans-7,8 dihydroxy-9,10-epoxy-7,8,9,-10-tetrahydrobenzo[a]pyrene (BPDE), was the best substrate. As for GSTA1-1, no obvious correlation between chemical reactivity or lipophilicity of the compounds and catalytic efficiencies was observed. Molecular modeling of diol epoxides in the active sites of GSTP1-1 and -A1-1 is in agreement with the assumption, based on functional studies, that the H-site of GSTA1-1 [Jernstrom et al. (1996) Carcinogenesis 17, 1491-1498] can accommodate stereoisomers of different sizes. Further, modeling of the enantiomers of anti- and syn-BPDE in the active site of GSTP1-1 provides an explanation for the exclusive preference for the enantiomers with R-configuration at the benzylic oxirane carbon. These isomers could be snuggly fitted in the H-site close to the GSH sulfur, whereas those with opposite stereochemistry could not. PMID- 9403174 TI - Biotransformation of the naturally occurring isothiocyanate sulforaphane in the rat: identification of phase I metabolites and glutathione conjugates. AB - Sulforaphane (SFN) is a naturally occurring isothiocyanate present in cruciferous vegetables, such as broccoli, that has been identified as a potent inducer of glutathione S-transferase activities in laboratory animals. The present studies were carried out to elucidate the metabolic fate of SFN in the rat. Particular emphasis was placed on glutathione (GSH)-dependent pathways because conjugation with GSH is a major route by which many isothiocyanates are eliminated in mammals. Male Sprague-Dawley rats were administered a single dose of SFN (50 mg kg-1 ip), and bile and urine were collected over ascorbic acid. Analysis of biological fluids was carried out by ionspray LC-MS/MS using the neutral loss (129 Da) and precursor ion (m/z 164) scan modes to detect GSH and N acetylcysteine (NAC) conjugates, respectively. In bile, five thiol conjugates (designated M1-M5) were detected. Metabolites M2 and M4 were identified as the GSH conjugates of SFN and erucin (ERN, the sulfide analog of SFN), respectively, by comparing their LC-MS/MS properties with those of standards obtained by synthesis. M1 was characterized as the GSH conjugate of a desaturated metabolite of SFN (tentatively assigned the structure of delta 1-SFN), suggesting that the parent compound also undergoes oxidative metabolism. Metabolites M3 and M5 were identified as the NAC conjugates of SFN and ERN, respectively, and together with the NAC conjugate of delta 1-SFN, these species also were detected in urine. Quantitative determination of the former two mercapturates in urine indicated that approximately 60% and approximately 12% of a single dose of SFN is eliminated in 24 h as the NAC conjugates of SFN and ERN, respectively. The corresponding figures in rats dosed with ERN were approximately 67% and approximately 29%. When the GSH conjugate of SFN was incubated with phosphate buffer (pH 7.4, 37 degrees C), < 1% of the conjugate dissociated to liberate free SFN. On the other hand, the conjugate underwent a facile thiol exchange reaction (> 70% conversion) when incubated in the presence of excess cysteine, thereby acting as an effective carbamoylating agent. It is concluded that SFN undergoes metabolism by S-oxide reduction and dehydrogenation and that GSH conjugation is the major pathway by which the parent compound and its phase I metabolites are eliminated in the rat. PMID- 9403175 TI - Substrate specificity of human O6-methylguanine-DNA methyltransferase for O6 benzylguanine derivatives in oligodeoxynucleotides. AB - To investigate the substrate specificity of human O6-methylguanine-DNA methyltransferase (MGMT) for O6-benzylguanine (6BG) derivatives incorporated in oligodeoxynucleotides, we prepared 25-mer lengths of sequences containing various 6BG derivatives and their related compounds and then measured the ability of these derivatives to inactivate MGMT in vitro. Oligodeoxynucleotides containing a 6BG, O6-(2-fluorobenzyl)guanine (2F-6BG), O6-(3-fluorobenzyl)guanine (3F-6BG), O6 (4-fluorobenzyl)guanine (4F-6BG), O6-benzylhypoxanthine (6BH), or O6 methylguanine (6MG) were all good substrates for MGMT, and no obvious differences were observed among them. Oligodeoxynucleotides containing N2-isobutyrylated 6BG and 6MG showed only a slightly reduced capacity for inactivating MGMT compared to N2-nonmodified forms of these derivatives. No obvious differences were observed in the corresponding double-stranded and single-stranded oligodeoxynucleotides. MGMT substrate specificity for the 6BG derivatives in the oligodeoxynucleotide was found to be quite different from that seen in our previous study [Mineura, K., et al. (1994) Int. J. Cancer 58, 706-712; (1995) Int. J. Cancer 63, 148-151. Kohda, K., et al. (1995) Biol. Pharm. Bull. 18, 424-430] and others [Moschel, R. C., et al. (1992) J. Med. Chem. 35, 4486-4491. Chae, M.Y., et al. (1994) J. Med. Chem. 37, 342-347] using the corresponding free bases. In brief, (i) 6BG, 3F-6BG, and 4F-6BG greatly inhibited human MGMT, whereas 2F-6BG, 6BH, and 6MG displayed much weaker activity; (ii) any modifications at the 2-amino group of the 6BG resulted in severe reductions in the ability to inactivate MGMT. These results obtained by the experiments using oligodeoxynucleotides and free bases suggest that human MGMT has low substrate specificity for 6BGs in oligodeoxynucleotides. Conformational changes in human MGMT which favor binding to oligodeoxynucleotides containing 6BG derivatives and the subsequent transfer of their benzyl groups may account for the difference in substrate specificity between the incorporated 6BG derivatives and their free base form. PMID- 9403176 TI - Hypochlorous acid-induced base modifications in isolated calf thymus DNA. AB - Exposure of calf thymus DNA to hypochlorous acid/hypochlorite leads to extensive DNA base modification. Large concentration-dependent increases in pyrimidine oxidation products [thymine glycol (cis/trans), 5-hydroxycytosine, 5 hydroxyuracil, 5-hydroxyhydantoin] but not purine oxidation products (8 hydroxyguanine, 2- and 8-hydroxyadenine, FAPy guanine, FAPy adenine) were observed at pH 7.4. In addition, large increases in 5-chlorouracil (probably formed from 5-chlorocytosine during sample preparation), a novel chlorinated base, were observed. Addition of HOCl to DNA already damaged by .OH generated by a mixture of ascorbate, copper(II) chloride, and hydrogen peroxide showed that hypochlorous acid led to a loss of 8-hydroxyguanine, 2- and 8-hydroxyadenine, FAPy guanine, FAPy adenine, and 5-hydroxycytosine in a concentration- and pH dependent manner. Nevertheless, time course studies suggested that the formation of purine oxidation products in isolated DNA by hypochlorous acid was not a major oxidation pathway. If this pattern of damage, especially the production of 5 chlorocytosine, is unique to hypochlorous acid, it might act as a "fingerprint" of damage to DNA by HOCl. PMID- 9403177 TI - Investigation of hydrolytic deamination of 1-(2-hydroxy-1-phenylethyl)adenosine. AB - The ring nitrogen of adenosine reacts at both the alpha- (benzylic) and beta carbons of styrene oxide to form 1-substituted products. The 1-(2-hydroxy-1 phenylethyl)adenosines formed by oxirane ring opening at the alpha-position are prone to an unusually facile hydrolytic deamination. By conducting hydrolysis reactions in [18O]water and analyzing the reaction products by electrospray mass spectrometry, we find that deamination occurs by direct attack of water at the 6 position of the adenine ring system with displacement of the exocyclic amino group. PMID- 9403178 TI - Effect of glutathione depletion on exocyclic adduct levels in the liver DNA of F344 rats. AB - The effects of glutathione (GSH) depletion on the in vivo formation of cyclic 1,N2- propanodexoxyguanosine adducts (AdG and CdG) as background lesions in the liver DNA of F344 rats were investigated. A group of 5 male F344 rats were given drinking water containing 30 mM L-buthionine (S,R)-sulfoximine (BSO) for 21 days, and another group of 8 rats were given only drinking water as controls. The BSO treated rats had significantly lower weight gain than control rats. The hepatic GSH levels in the BSO-treated group were reduced by 84% as compared with the control group, from 4.43 to 0.72 mumol/g of tissue. The isomeric AdG3, CdG1, and CdG2 were detected by the 32P-postlabeling/HPLC method in the liver DNA of rats without carcinogen treatment, as we reported previously [Nath, R. G., and Chung, F.-L. (1994) Proc. Natl. Acad. Sci. U.S.A. 91, 7491-7495. Nath, R. G., et al. (1996) Cancer Res. 56, 452-456]. The mean levels (mumol/mol of guanine) for AdG3, CdG1, and CdG2 were 0.57 +/- 0.25, 0.15 +/- 0.18, and 0.16 +/- 0.22 for the control group and 1.18 +/- 1.03, 3.16 +/- 3.26, and 2.50 +/- 2.59 for the BSO group, respectively. These increases correspond to approximately 2-fold for AdG and 15-21-fold for CdG adducts. The dramatic increase in the cyclic adduct levels in rat liver DNA could have resulted mainly from GSH depletion as a result of the BSO treatment, even though other unknown effects due to the toxicity of BSO cannot be ruled out. These results suggest that GSH plays an important role in protecting the liver against cyclic propano DNA adduction and provide further support for the endogenous origin of these adducts. PMID- 9403179 TI - Synthesis and cleavage of oligodeoxynucleotides containing a 5-hydroxyuracil residue at a defined site. AB - Oxidation and hydrolysis of a cytosine residue can lead to the formation of 5 hydroxyuracil in DNA. The biological consequences of this modification are not fully understood. To facilitate biochemical and biophysical studies aimed at elucidating the effects of this modification in DNA, we have developed a solid phase synthetic method for the placement of 5-hydroxyuracil residues at defined sites in oligodeoxynucleotides. This method is based upon the enhanced acidity of the 5-hydroxyl proton which allows selective aqueous acetylation. Under standard aqueous ammonia deprotection conditions, however, we observed that 5 hydroxyuracil residues are lost substantially from synthetic oligonucleotides. Substitution of aqueous ammonia with methanolic potassium carbonate and the use of phosphoramidite derivatives with alternatively protected amino groups allow synthesis of oligonucleotides containing 5-hydroxyuracil and all normal bases in high yield. The composition of the oligodeoxynucleotides prepared by this method has been verified by enzymatic digestion followed by high-performance liquid chromatography (HPLC) analysis as well as acid hydrolysis followed by GC/MS analysis. The location of the 5-hydroxyuracil residue is demonstrated by selective permanganate oxidation of the 5-hydroxyuracil residue followed by beta elimination. We have also probed a synthetic oligonucleotide containing a unique 5-hydroxyuracil residue with uracil DNA N-glycosylase, previously reported to remove this lesion from DNA. PMID- 9403180 TI - Synthesis and 32P-postlabeling/high-performance liquid chromatography separation of diastereomeric 1,N2-(1,3-propano)-2'-deoxyguanosine 3'-phosphate adducts formed from 4-hydroxy-2-nonenal. AB - 4-Hydroxy-2-nonenal (HNE), a major electrophilic byproduct of lipid peroxidation, is mutagenic and cytotoxic. The two pairs of HNE-derived diastereomeric 1,N2 propanodeoxyguanosine 3'-monophosphate adducts were synthesized from reaction of HNE with 2'-deoxyguanosine 3'-monophosphate. After HPLC separation, these adducts were characterized by UV-visible absorption and negative ion electrospray ionization MS/MS analysis. To further characterize the structures, these adducts were dephosphorylated to the corresponding HNE-modified deoxyguanosine adducts and their HPLC retention times and UV spectra were compared with those of the synthetic standards prepared from reaction of HNE with 2'-deoxyguanosine. Separation of these adducts by 32P-postlabeling/HPLC was developed. Reaction of HNE with calf thymus DNA resulted in only one pair of diastereomeric adducts, with one adduct predominantly formed with a modification level of 1.2 +/- 0.5 adducts/10(7) nucleotides. PMID- 9403181 TI - Effect of substitution site upon the oxidation potentials of alkylanilines, the mutagenicities of N-hydroxyalkylanilines, and the conformations of alkylaniline DNA adducts. AB - Carcinogenic arylamines typically undergo metabolic activation via N hydroxylation followed in most instances by O-esterification. In this study, the ability of methyl-, dimethyl-, and ethylaniline constituents of tobacco smoke to undergo oxidation at the nitrogen atom was analyzed. In addition, the mutagenicity of the corresponding N-hydroxyalkylanilines and the conformational properties of the DNA adducts generated from their N-acyloxy derivatives were investigated. All the arylamines underwent irreversible electrochemical N oxidation at potentials higher than those observed for the oxidation of carcinogenic polynuclear aromatic amines. There were minor differences in the oxidation potentials, which were consistent with the position and electron donating abilities of the alkyl substituents; however, the differences appeared to be too small to account for the range of genotoxic effects among the alkylanilines. N-Hydroxyarylamines containing p-alkyl substituents had increased mutagenicities in Salmonella typhimurium TA100, which was attributed to their higher efficiencies of adduct formation. Increased mutagenicities were also observed upon o-alkyl substitution; however, this property was not related to a greater ability of the ortho-substituted species to form DNA adducts, which suggested that adducts from ortho-substituted alkylanilines may be intrinsically more mutagenic than their meta- and para-substituted analogues. In all instances, N-(acyloxy)-arylamines generated from the N-hydroxyarylamines reacted with dG, dG nucleotides, and DNA to yield C8-substituted dG derivatives as the major adducts. The alkylaniline-dG adducts displayed distinct conformational trends that were determined by the location of the alkyl substituents. Spectroscopic data indicated higher percentages of low-energy syn conformers in the adducts that contained alkyl groups ortho to the arylamine nitrogen as opposed to adducts not bearing ortho substituents. The data strongly suggest that the conformational properties of the DNA adducts, in particular their ability to adopt syn conformations, may be determinant factors for the genotoxic responses elicited by certain alkylanilines (e.g., 2-methylaniline and 2,6-dimethylaniline). PMID- 9403182 TI - Covalent DNA adducts formed by benzo[c]chrysene in mouse epidermis and by benzo[c]chrysene fjord-region diol epoxides reacted with DNA and polynucleotides. AB - The metabolic activation in mouse skin of benzo[c]chrysene (B[c]C), a weakly carcinogenic polycyclic aromatic hydrocarbon (PAH) present in coal tar and crude oil, was investigated. Male Parkes mice were treated topically with 0.5 mumol of B[c]C, and DNA was isolated from the treated areas of skin at various times after treatment and analyzed by 32P-postlabeling. Seven adduct spots were detected, at a maximum level of 0.89 fmol of adducts/microgram of DNA. Four B[c]C-DNA adducts persisted in skin for at least 3 weeks. Treatment of mice with 0.5 mumol of the optically pure putative proximate carcinogens (+)- and (-)-trans-benzo[c]chrysene 9,10-dihydrodiols [(+)- and (-)-B[c]C-diols] led to the formation of adducts which comigrated on TLC and HPLC with some of those formed in B[c]C-treated mice. The major adduct formed in mouse skin treated with B[c]C coeluted on TLC and HPLC with an adduct formed in mouse skin treated with (-)-B[c]C-diol. These results suggested that the detected adducts were formed by the fjord-region B[c]C-9,10 dihydrodiol 11,12-epoxides (B[c]CDEs). To test this, the four optically pure synthetic B[c]CDEs were reacted in vitro with DNA and with synthetic polynucleotides and these samples were 32P-postlabeled. Cochromatography, both on TLC and HPLC, of in vitro and in vivo adducts indicated that B[c]C is activated in mouse skin through formation of the (-)-anti- and (+)-syn-B[c]CDE with 9R,10S,11S,12R- and 9S,10R,11S,12R- absolute configuration, respectively, both of which formed two DNA adducts in vivo. However, the major adduct present in the B[c]C-treated skin DNA was not a fjord-region B[c]CDE adduct but was possibly derived from a bay region B[c]CDE at the 1,2,3,4-position. The extent of DNA adduct formation by B[c]C in mouse skin DNA was lower than that of moderately carcinogenic PAHs previously studied by this method, suggesting a correlation between extent of DNA adduct formation and carcinogenic potential. PMID- 9403183 TI - Formation and properties of peroxynitrite as studied by laser flash photolysis, high-pressure stopped-flow technique, and pulse radiolysis. AB - Flash photolysis of alkaline peroxynitrite solutions results in the formation of nitrogen monoxide and superoxide. From the rate of recombination it is concluded that the rate constant of the reaction of nitrogen monoxide with superoxide is (1.9 +/- 0.2) x 10(10) M-1 s-1. The pKa of hydrogen oxoperoxonitrate is dependent on the medium. With the stopped-flow technique a value of 6.5 is found at millimolar phosphate concentrations, while at 0.5 M phosphate the value is 7.5. The kinetics of decay do not follow first-order kinetics when the pH is larger than the pKa, combined with a total peroxynitrite and peroxynitrous acid concentration that exceeds 0.1 mM. An adduct between ONOO- and ONOOH is formed with a stability constant of (1.0 +/- 0.1) x 10(4) M. The kinetics of the decay of hydrogen oxoperoxonitrate are not very pressure-dependent: from stopped-flow experiments up to 152 MPa, an activation volume of 1.7 +/- 1.0 cm3 mol-1 was calculated. This small value is not compatible with homolysis of the O-O bond to yield free nitrogen dioxide and the hydroxyl radical. Pulse radiolysis of alkaline peroxynitrite solutions indicates that the hydroxyl radical reacts with ONOO- to form [(HO)ONOO].- with a rate constant of 5.8 x 10(9) M-1 s-1. This radical absorbs with a maximum at 420 nm (epsilon = 1.8 x 10(3) M-1 cm-1) and decays by second-order kinetics, k = 3.4 x 10(6) M-1 s-1. Improvements to the biomimetic synthesis of peroxynitrite with solid potassium superoxide and gaseous nitrogen monoxide result in higher peroxynitrite to nitrite yields than in most other syntheses. PMID- 9403184 TI - Preoperative pulmonary evaluation of the thoracic surgical patient. AB - A test designed to separate those undergoing thoracic surgery without complications and those with complications must be both highly specific and sensitive. Clearly, the difference between patients at opposite ends of the population curves is easy to identify. Spirometry can be helpful for screening, although it is not a very discriminating test. If patients fall in the overlap region between the populations, however, it is impossible to discern the risks with any certainty using low-yield tests. A test with higher sensitivity, specificity, and predictive values is necessary to ascertain such marginal differences. With this kind of analysis at hand, preoperative testing can be divided into three predictive value groups. Calculating the predictive value of each preoperative test can provide a comparative measure of usefulness of discriminative power (Table 1). In this way, spirometry, blood gas analysis, and stair climbing tolerance are shown to be poor predictors of outcome. An intermediate predictive value can be achieved using diffusion capacity, exercise induced decreases in O2 saturation, and exercise PVR. High predictive value can be accomplished with combination indexes (PPP, possibly PRQ), measurement of VO2 at 40 watts of exercise, or VO2max. Logic dictates a step-wise preoperative evaluation using prediction value analysis (Fig.4). A flow decision chart for the preoperative evaluation of patients for pulmonary resection begins with exercise oximetry, spirometry, and blood gas analysis as general screening tests to separate those patients at minimal or no risks for complications from those patients that require further evaluation. Functional indexes (PPP, PRQ) or exercise testing can aid further in the selection of those patients in whom a nonsurgical option should be considered. Flow decision chart for the preoperative evaluation of patients for pulmonary resection should continue to evolve as new information about outcome studies is gathered. Examination of outcome data will provide us with reduction of the size of the nonoperable population, so that we can deny only those patients who truly pose a prohibitive risk. PMID- 9403185 TI - Preoperative cardiac assessment of the thoracic surgical patient. AB - The preoperative cardiac assessment of the thoracic patient differs very little from the assessment of any patient for noncardiac surgery, aside from a few special issues. Therefore, rather than reviewing the general issue of evaluation for noncardiac surgery, which is a topic that has been reviewed many times in the recent past, this article focuses on the purposes, methods, and limitations of risk assessment in the noncardiac surgical patient with suspected coronary artery disease (CAD), including thoracic surgical patients. Because risk assessment is imprecise and the main indications for invasive coronary procedures prior to noncardiac surgery are the same for any person for whom life-expectancy is expected to be prolonged, meticulous preoperative evaluation for CAD is not usually warranted, even for patients undergoing high-risk surgery or with multiple risk factors for CAD. To help understand this point of view and to utilize cardiology consultants appropriately, implications of basic pathophysiology as well as statistical principles are also discussed. PMID- 9403186 TI - Pulmonary rehabilitation exercise program for high-risk thoracic surgical patients. AB - Preoperative assessment of the patient with moderate to severe COPD continues to be a difficult task. Pulmonary function tests cannot be the only deciding factor. Exercise testing is supported in the literature to improve the sensitivity and specificity in predicting perioperative morbidity and mortality rates. The presence of comorbid disease, especially COPD, increases the chances of postoperative pulmonary complications. Pulmonary rehabilitation in the nonsurgical COPD patient has been proven to be beneficial in improved exercise capacity and quality of life. The question which remains to be answered is whether a short-term, intense, and focused preoperative program in surgical candidates with resectable NSCLC and preexisting pulmonary dysfunction can influence outcome. PMID- 9403187 TI - Selection of anesthetic agents for thoracotomy. AB - The selection of anesthetic agents for thoracotomy requires consideration of multiple factors. The goals of anesthetic management are to choose an anesthetic plan that will allow not only for appropriate intraoperative conditions but also for rapid extubation and adequate pain control so that the perioperative course will have the least possible associated risk of morbidity and mortality. A patient's pre-existing medical condition must be optimized. Length and type of surgical procedure, as well as the patient's medical status, determine anesthetic technique and agents. PMID- 9403189 TI - Lung isolation. Tube design and technical approaches. AB - Airway management for thoracic surgery frequently requires isolation of a portion of the respiratory system. In some circumstances lung isolation is mandatory and in others elective. Several techniques utilizing specialized endotracheal tubes and blockers are currently available. There are specific advantages and complications associated with each that, in part, determine optimal outcome in this specialized group of surgical patients. PMID- 9403188 TI - Intraoperative monitoring. AB - This article discusses some of the routine as well as more specialized monitoring devices available. In thoracic surgery monitoring may be even more challenging because the surgery itself may involve manipulation of the airways, the pulmonary as well as cardiovascular systems. The anesthesiologist must have a full understanding of the required monitoring devices and decide which if any special techniques are needed depending on the surgical procedure and the patient's preoperative condition. PMID- 9403190 TI - Physiology of the lateral position and one-lung ventilation. AB - The first part of this article reviews the distribution of ventilation (V) and perfusion (Q) during the supine and the lateral decubitus position. The changes in the V/Q during the lateral position with and without paralysis are discussed. The second part evaluates the degree of transpulmonary shunt during one lung ventilation (OLV) and the role of hypoxic pulmonary vasoconstriction in maintaining arterial oxygenation. Finally, the influence and the use of nitric oxide during OLV is reviewed. PMID- 9403191 TI - Postoperative pain management. AB - Postoperative pain management is essential and must be approached as an integral part of the perioperative care. It should be systematic and based on sound physiologic and pharmacologic principles. The intra-operative management of pain is crucial, as there is perhaps an important role for preemptive analgesia. Because of its unique nature, pain is difficult to assess, but for good results adequate and repeated assessment are vital. The literature also points to the detrimental effects of inadequate pain control. There are a variety of methods available for pain management. In choosing a method, various factors need to be considered including physician skill, knowledge of analgesics and routes of administration, patient-related and clinical circumstances, the availability of an environment supportive of effective pain management, and the knowledge and skill of staff to assess and monitor patients. These need to be considered along with the risks and benefits and cost-benefit of the various drugs and techniques. The cornerstone of therapy is opioids, which can be administered by a variety of routes. The use of TEA with opioids and local anesthetics is highly beneficial, especially in high-risk patients. The aim should be to provide all patients a balanced analgesic regimen based on the identification of multiple mechanisms involved in postoperative pain. PMID- 9403192 TI - Mechanical ventilation of the postoperative patient. AB - This article explains the operating characteristics of the most commonly used modes of mechanical ventilation and provides descriptions of how mechanical breaths are delivered. Also included are simple guidelines for choosing tidal volume, respiratory and flow rate, and alarm settings. The article concludes with a brief overview of weaning techniques and commonly used criteria to determine when patients are ready to be weaned from ventilating support. PMID- 9403193 TI - Prevention and management of dysrhythmias following thoracic surgery. AB - Supraventricular dysrhythmias (SVDs) occur frequently after thoracic surgery and have been associated with prolonged hospital stays. The reported incidence of supraventricular dysrhythmias in the thoracic surgery patient population ranges from 10% to 40%, with factors such as age and extent of surgery markedly influencing the incidence. This article focuses on new issues leading to improved understanding of the pathophysiology and mechanisms of SVDs after surgery. New approaches directed at prophylaxis and acute therapy of SVDs are also discussed. PMID- 9403194 TI - Anesthesia for the pediatric patient. AB - Infants and children have unique anatomic, physiologic, pharmacologic, and psychological issues relating to perioperative management. Combining this knowledge with the technical skills required for instrumentation of children is essential when contemplating anesthesia for thoracic surgery. Experience and versatility with anesthetic induction technique, airway instrumentation, vascular access and monitoring, single-lung ventilation, regional anesthesia, and postoperative pain management allow for the comprehensive management of thoracic surgical patients at any age. PMID- 9403195 TI - Aqueduct stenosis in animal models of hydrocephalus. PMID- 9403196 TI - MRI in Smith-Lemli-Opitz syndrome type 1. PMID- 9403197 TI - Primary intracranial neoplasms of infancy and early childhood. AB - We investigated the age-related location, gender distribution, and histology of 107 brain tumors in children under 4 years of age seen in our department between 1984 and 1997. The male-to-female ratio was 1.4 (62/45 cases) with a prevalence of supratentorial tumors (60/47 = 1.3); the main histological entity was astrocytoma (33.6%), followed by ependymoma (14.0%). In the 1st year of life 22 cerebral neoplasms became clinically apparent. A higher ratio for supratentorial tumors was revealed (17/5 = 3.4), but without gender preference, and primitive neuroectodermal tumors (PNET) were the most frequent (5/22). In the 2nd year 25 tumors were found. The male-to-female ratio was 1.5 (15/10) and the supratentorial-to-infratentorial ratio, 1.1 (13/12). The two most common entities were astrocytoma and ependymoma (6 cases each). In addition, a survey of previously published investigations into this subject was performed and a compilation of data on 1960, 545 and 1084 tumors in children below the age of 1, 2 and 4 years, respectively, was prepared, which makes it the most extensive review of brain tumors of infancy and early childhood yet undertaken. PMID- 9403198 TI - Thalamic tumors in children. AB - Thalamic tumors (TT) merit individual analysis and must not be confused with tumors that, while involving the entire thalamus have a different origin. We analyzed 26 patients who fulfilled our criteria of having "strictly" TT. We examined incidence, clinical features, histology, response to treatment (mainly surgery), recurrence rate, mortality and prognosis. We considered that histology and surgical treatment were the most important items related to prognosis. Low grade tumors (LGT) had a good prognosis, while anaplastic tumors (AT) had a discouraging one; nevertheless both must be operated on. We believe that total removal of LGT is curative and total removal of AT, even if it is not curative, can extend survival by some months. Radiotherapy and chemotherapy seemed to be of little value in our series of TT. PMID- 9403199 TI - Cerebral cavernous hemangiomas in childhood. Clinical presentation and therapeutic considerations. AB - Cerebral cavernous hemangiomas (CCH) are relatively rare vascular hamartomas. Since the introduction of MRI there has been an increase in the number of case reports of CCH in the medical literature. CCH are often asymptomatic; they may, however, cause epilepsy or neurological deficits due to their space-occupying effects or hemorrhagic sequelae. The tendency of CCH to bleed has been well recognized, though gross hemorrhage is infrequent owing to the relatively low blood pressure and small blood flow in CCH. MRI findings of a CCH are characteristic and can differentiate the lesions from other vascular abnormalities. To date, there has been no consensus on indications for surgical intervention. Three cases are presented, which together demonstrate by their different presentation, clinical course and MRI findings that each patient with a CCH requires an individually tailored management. Presentation, clinical course and accessibility for operation are the factors that determine whether a surgical or a conservative approach should be adopted. PMID- 9403200 TI - Continuous external subdural drainage in the management of infantile subdural collections: a prospective study. AB - Continuous external subdural drainage (CESD) was suggested as a treatment step to be inserted prior to SP shunting, primarily because it makes it possible to avoid shunt placement in a significant number of patients. Thirty-three patients with symptomatic chronic subdural collection confirmed by computed tomography were included in this study. Unilateral CESD was performed in all cases, using a lumbar drainage set. The drains were left in place for no more than 10 days. A subduroperitoneal (SP) shunt was inserted in those patients in whom re accumulation of the subdural collection had occurred. Of 33 patients, 17 were definitively treated by CESD and 16 subsequently needed an SP shunt. The cost of treatment with CESD was just less than half the cost of treatment with SP shunting. CESD can be used as a step before SP shunting in the management of chronic infantile subdural collections, since it is effective without further treatment in half the patients and safer than subdural tapping. PMID- 9403201 TI - Outcome and life prospects after surgical management of medically intractable epilepsy in patients under 18 years of age. AB - A retrospective analysis of seizure outcome and quality of life assessment was done in 64 patients under 18 years of age with medically refractory epilepsy who underwent 64 primary and 16 repeat operative procedures in an attempt to control their epilepsy. At least 2 years' follow-up data were available for each patient. Operative procedures were 44 temporal lobe resections; 16 extratemporal resections; and 4 hemispherectomies. Effective control of previously intractable seizures was obtained in most patients: 55%, 11%, and 17% achieved Engel class I, II, and III status, respectively. Successful seizure control was thus obtained in 83%, while 17% (Engel class IV) failed to improve significantly after operation. Quality-of-life measures parallelled the improvements in seizures control, being highest in Engel I, outcome group and lowest in Engel IV outcome group. In appropriately selected pediatric and adolescent patients with medical refractory epilepsy, surgical management can offer a safe and effective adjunct to medication. PMID- 9403202 TI - Tumors of the orbit. Pitfalls of the surgical approach in 37 children with orbital tumor. AB - The authors review their experience with 37 children with orbital tumors, summarizing their surgical techniques, the indications applied, and the pitfalls of each surgical approach. Tumors located in the retro-ocular or intraorbital space were surgically excised through a transcranial approach (28 cases), while for tumors in other sites lateral orbitotomy (5 cases), medial orbitotomy (1 case) and biopsy (3 cases) were performed. A transcranial approach was used for tumors with intracranial extension and for those located in the orbital apex and deep medial orbital compartment. Lateral orbitotomy was used for tumors located in the superior, temporal or inferior compartment of the orbit and those in the lateral apex. A medial orbitotomy was used for tumors located medial to the optic nerve. Outcomes of the surgical intervention varied, depending on the pathology, location and extent of the individual tumors. To obtain optimal exposure and minimize functional deficits, the pitfalls of surgical approaches to orbital tumors are discussed. PMID- 9403203 TI - Acute cerebellar ataxia in children. AB - Acute cerebellar ataxia is a benign syndrome usually occurring after an acute febrile disease. In a few cases neuroradiological investigations reveal cerebellar alterations. Clinical and neuroradiological involvement of the brain stem has rarely been reported in the literature. We present five cases of acute cerebellar ataxia. In two cases the cerebellar symptomatology was associated with neurological signs of brain stem involvement. CT scans did not show any pathologic findings in three patients. MRI disclosed cerebellar or brain stem alterations in all the patients. Clinical and neuroradiological findings allow differentiation of this pathologic entity from other demyelinating or dysmyelinating diseases. The value of MRI in detection and localization of the lesions and in following their evolution is emphasized. PMID- 9403204 TI - Immunoglobulin prophylaxis in shunt infections: a prospective randomized study. AB - Cerebrospinal fluid shunt infection is serious and one of the most frequent complications of shunt implantation. Age has been one of the most significant host factors for the development of shunt infections. A relative deficiency of the immune response against bacteria in infants could partly explain the higher infection rate in the very young patients. This prospective-randomized study was conducted in two groups: group A (immunoglobulin group) and group B (control group). There were 30 patients in each group. The patients in group A received intravenous immunoglobulin (Sandoglobulin) at a dose of 1 g/kg in the night before surgery. Each patient was followed up to 6 months. No infection was seen in group A. In group B, infection rate per procedure were 5.1% (P = 0.494) and 6.6% (P = 0.492), respectively. Intravenous immunoglobulin prophylaxis in infants seems to reduce the shunt infections. PMID- 9403205 TI - Intracranial and spinal metastases from a ganglioglioma with unusual cytogenetic abnormalities in a patient with complex partial seizures. AB - We describe an unusual clinical presentation of a ganglioglioma in a patient with complex partial seizures. The patient underwent a right temporal lobectomy with subtotal tumor resection at age 15 years, followed by a complete resection 1 year later. Follow-up MRI scan a year later documented recurrence and leptomeningeal dissemination. Another biopsy was performed. Pathological examination revealed similar histology in all three resections, with a ganglioglioma showing no evidence of anaplasia. The tumor exhibited a number of karyotypic abnormalities, notably, a paracentric inversion of chromosome 7. In summary, despite lacking anaplastic features by conventional histological criteria, this ganglioglioma showed an unsusual karyotype and demonstrated radiological evidence of widespread dissemination. PMID- 9403206 TI - Congenital immature teratoma of the fetal brain. AB - Congenital intracranial tumors are very rare and only account for 0.5-1.5% of all childhood brain tumors. Even rarer are those with prenatal manifestation. The most common of these present at birth are teratomas, which show divergent differentiation with 90% of them containing tissues from all three germ layers. We report a rare case of an intrauterine congenital immature teratoma in a female fetus at 23 weeks of gestation, which was sonographically diagnosed in vivo by detection of the tumor and associated craniomegaly. Because of the poor prognosis, termination of the pregnancy was induced by Rivanol instillation. The cerebral tumor was confirmed at autopsy and was not associated with any other malformations. Histological and immunohistochemical features of this tumor are presented. PMID- 9403207 TI - Assessment of brain perfusion by 99mTc-HMPAO SPECT in akinetic mutism due to high dose intravenous methotrexate therapy. AB - Chemotherapy of the central nervous system may cause neurotoxicity in children with acute lymphocytic leukemia. We evaluated regional blood flow in a 6-year-old child presenting with akinetic mutism, using 99mTc-HMPAO single photon emission tomography (SPECT) following high-dose intravenous methotrexate therapy. While findings in X-ray computerized tomography were decreased density in bilateral basal ganglia and thalamic nuclei with diffusely decreased attenuation of the periventricular white matter, a global, frontal dominant profoundly abnormal perfusion pattern involving both gray and white matter was observed in the SPECT study. Treatment of the central nervous system with high dose intravenous chemotherapy may cause profound abnormalities in white and gray matter blood flow and early assessment of the neurotoxicity may be identified by 99mTc-HMPAO SPECT in the pediatric age group. PMID- 9403208 TI - Germinoma in cerebral hemisphere associated with Down syndrome. AB - A Down syndrome patient with germinoma developing in the cerebral hemisphere is reported. A review of the literature yielded only 14 cases of Down syndrome with brain tumors, including our case. This finding of brain tumors in patients with Down syndrome may reflect chance occurrence. However, it is of interest in this regard that in 6 of the 14 (43%) reported cases the lesions were intracranial germ cell tumors. PMID- 9403209 TI - Rapidly progressive IgA nephropathy with anti-myeloperoxidase antibodies benefits from immunosuppression. AB - CLINICAL OBSERVATIONS: Three patients with previous pulmonary infections were recently admitted with rapidly progressive renal failure. Renal biopsy showed crescentic glomerulonephritis with deposits of IgA, C3c and C3d. Serology disclosed P-ANCA with high-titer anti-myeloperoxidase antibodies. Two out of three patients became dialysis dependent despite immunosuppression with methylprednisolone and cyclophosphamide. Renal function improved in both patients after 2 weeks and 9 months, respectively, permitting termination of hemodialysis. All patients benefited from immunosuppressive treatment which is currently still being continued. CONCLUSION: The data suggest that early immunosuppression is beneficial in patients presenting with crescentic rapidly progressive IgA GN and anti-myeloperoxidase antibodies, which may represent a novel subset of crescentic IgA GN associated with high-titer anti-myeloperoxidase antibodies constituting an overlap group between microscopic polyangiitis and IgA GN. PMID- 9403210 TI - Membranoproliferative glomerulonephritis induced by portosystemic shunt surgery for non-cirrhotic portal hypertension. AB - The liver and spleen both have important phagocytic functions and contain monocytes/macrophages which clear immune complexes. We describe here three patients who presented proteinuria and hematuria 7 to 13 years after portosystemic shunt surgery, which diverted portal venous blood to the systemic circulation. They had hematemesis and/or melena and underwent mesocaval shunt surgery and splenectomy in childhood because of non-cirrhotic portal hypertension with esophageal varices. Renal biopsy specimens revealed findings characteristic of membranoproliferative glomerulonephritis (MPGN) type I. Immunohistologically, these three cases were accompanied by a distinct IgA deposition along with a marked C3 deposition. The IgA observed in these three cases contained not only IgA1 but also IgA2, which is the predominant form of mucosal IgA. On the other hand, of 20 patients with idiopathic MPGN type I with IgA deposition (n = 20), only two were positive for IgA2, and the distribution was focal and segmental. Our study shows that MPGN type I may have developed secondary to portosystemic shunt. This secondary form of MPGN type I may be caused by a reduced clearance of immune complexes in the liver and their deposition in the glomerulus, since a portosystemic shunt routes portal venous blood from the intestinal tract directly to the systemic circulation. PMID- 9403211 TI - Plasmapheresis in the treatment of steroid-resistant focal segmental glomerulosclerosis. AB - A patient presented with a severe nephrotic syndrome and a renal biopsy consistent with early focal segmental glomerulosclerosis (FSGS). After a year of intensive immunosuppressive therapy proteinuria was unabated and renal function began to deteriorate. Treatment with weekly plasmapheresis combined with moderate doses of prednisone and azathioprine produced a dramatic decrease in proteinuria and serum creatinine. A marked fall in the B-cell population occurred during treatment, as well as an increase both in T-lymphocytes of the mature CD4+ helper/suppressor phenotype and in the immature/cytotoxic CD8+ phenotype. Activation of the immune system during treatment was demonstrated by an increase in the rate of spontaneous proliferation of peripheral blood mononuclear cells and an increase in T-cell expression of the interleukin-2 receptor. PMID- 9403212 TI - Hematuria due to hyperuricosuria in children: 36-month follow-up. AB - Hyperuricosuria (HU), defined as a urinary acid excretion higher than 95 percent of normal values, is an important lithogenic factor, accounting for about 5-20% of recurrent hematuria in childhood. We prospectively studied 30 children (15 male, 15 female; aged 3 to 13 years old) with previously undiagnosed isolated hematuria and HU for 6 to 36 months. The family history of nephrolithiasis was positive in 40%. Idiopathic hypercalciuria (IH), UCa > 4 mg/kg/day, was not found initially, but was diagnosed after 6 to 24 months in 20% of the patients. The following treatments were utilized: restricted purine diet (13%), allopurinol (4%) and potassium citrate (1%). No specific treatment was given to 82% of the patients. Therapy normalized uricosuria with resolution of hematuria over 6-12 months. Thirteen percent and 6% of untreated patients developed urolithiasis after 6 and 12 months respectively. The data suggest that HU, like IH, is associated with hematuria. Furthermore, recognition of this association may prevent unnecessary, and in some cases, invasive diagnostic manoeuvres. PMID- 9403213 TI - Validation by image analysis of a turbidimetric method to study calcium oxalate crystallization. AB - Numerous studies of calcium oxalate crystal formation have been carried out in the past two decades. In the present study, experiments were carried out to validate a turbidimetric method allowing to assess the calcium oxalate crystallization process. This method is quick and reproducible and can be used to quantify the inhibition of calcium oxalate crystal growth by various compounds. An experimental method of validation has been developed, which consisted in filtering solutions pure or containing modifiers at given crystallization times, photographing the filters used on scanning electron microscopy and analyzing the images using mathematical methodology. The results obtained through image analysis, namely crystal density (mean particle number per unit volume) and mean area, were correlated with the turbidimetric parameters. This finding was consistent with the qualitative examination of the photographs. Moreover, the morphological differences in crystals observed on the photographs were confirmed by the calculated length/width ratio. One can therefore assume that inhibition of calcium oxalate crystal growth is at least, partly explained by surface adsorption phenomena, which may add to complex formation. PMID- 9403214 TI - Fate of recurrent acute interstitial cellular rejection in an HLA identical kidney transplant recipient: impact of donor microchimerism. AB - We and others have shown that the incidence of acute interstitial rejection in HLA identical and non-identical kidney transplantation is similar. Chronic vascular rejection is, however, rare in full matched recipients. In light of the known correlation between previous acute cellular and chronic vascular rejections, lack of chronic rejection in full matched kidney allografts suggest that the immunological basis of acute and chronic rejections are different and/or only high-grade acute cellular rejection leads to chronic vascular rejection. Herein, we present the case of an HLA identical kidney transplant from a male donor to a female recipient who, because of poor compliance, had frequent acute interstitial cellular rejection culminating into chronic interstitial fibrosis with no evidence of vasculopathy or glomerulopathy characteristic of chronic vascular rejection. Donor cells, as examined by the presence of the Y chromosome DNA, were present in the peripheral blood during the most recent acute rejection but not thereafter. These findings support the notion that acute interstitial cellular rejection can lead to interstitial fibrosis but not chronic vasculopathy/glomerulopathy, and that microchimerism did not confer protection against acute cellular rejection. PMID- 9403215 TI - Effects of isradipine on renal hemodynamics in renal transplant patients treated with cyclosporine. AB - Cyclosporine A is the mainstay of modern immunosuppressant therapy in human organ transplants. However, its use has been associated with nephrotoxicity and hypertension that may be hemodynamically mediated. This study was undertaken to see if the calcium channel blocker, isradipine, could improve renal hemodynamics by decreasing renal vascular resistance and to determine its effects on cyclosporine pharmacokinetics. Eighteen hypertensive renal transplant recipients (3 to 60 months post transplant) were enrolled. Antihypertensive medications were stopped. Patients were placed on isradipine and followed for 8 weeks. Blood pressure, creatinine clearance, cyclosporine trough levels, and renal plasma flow using p-aminohippurate clearance, were measured pre- and at 8 weeks of therapy. Only one patient did not complete renal plasma flow measurement. Systolic and diastolic blood pressures dropped significantly (p < 0.001) from baseline to 8 weeks with isradipine. Sbp decreased from 166 +/- 18 to 142 +/- 14 mmHg and Dbp decreased from 97 +/- 8 to 82 +/- 10 mmHg (mean +/- 1 SD). Renal vascular resistance, calculated from mean arterial pressure and renal blood flow, decreased significantly (p < 0.002) from 0.57 +/- 0.21 to 0.41 +/- 0.12 mmHg/ml/min (mean +/- 1 SD). Isradipine appeared to have no significant effect on cyclosporine trough levels, creatinine clearance, or renal plasma flow. This study shows a role for isradipine use in renal transplant recipients on cyclosporine. Renal vasoconstriction caused by cyclosporine may be partially prevented with isradipine therapy. Furthermore, isradipine is effective in the treatment of hypertension in renal transplant recipients without affecting cyclosporine levels. PMID- 9403216 TI - A case of superantigen-related glomerulonephritis after methicillin-resistant Staphylococcus aureus (MRSA) infection. AB - A case in which the enterotoxins of Staphylococcus aureus may have served as bacterial superantigens is presented. This 71-year-old man developed proteinuria and renal dysfunction after contacting pneumonia caused by methicillin-resistant Staphylococcus aureus (MRSA), coagulase type II. The infection occurred after surgery for recurrent lung cancer. Staphylococcus enterotoxins B, C, and TSST-1 were detected from the bacillus. Ten days after the onset of pneumonia, proteinuria was noted; urinary protein was as high as 1.8 g/day. The serum creatinine was elevated from 1.0 mg/dl to 3.7 mg/dl. Several immunological reactions were detected; the serum levels of IgG and IgA were increased, and the selective usage of T-cell receptor V beta (TCRV beta) was observed. Serum levels of IL-1 beta, IL-2, IL-6, IL-8, IL-12, and tumor necrosis factor alpha (TNF alpha) were also elevated. Examination of the renal biopsy specimen by light microscopy showed minor to mild mesangial proliferative glomerulonephritis. Immunofluorescence microscopy demonstrated the deposition of IgG, IgA, and C3, mainly along the capillary walls. Electron microscopy revealed electron dense deposits, mainly in the subepithelial areas, and injury to the glomerular basement membrane. When the pneumonia improved following antibiotic therapy, the renal function also improved, and proteinuria decreased. The levels of immunoglobulins and the usage of TCRV beta also decreased. Because staphylococcus enterotoxins act as superantigens, we believe this to be a typical case of superantigen-related glomerulonephritis. PMID- 9403217 TI - Cerebral venous thrombosis in nephrotic syndrome. AB - This report describes cerebral venous sinus thrombosis, a rare and perhaps under diagnosed complication of nephrotic syndrome. We review the pathophysiology of the coagulopathy associated with nephrotic syndrome including abrupt renal loss of antithrombin III. We propose a rationale approach to treating this condition with low-molecular-weight heparin and antithrombin III replacement. PMID- 9403218 TI - Lobular form idiopathic glomerulonephritis with massive subendothelial and paramesangial immune deposits, a three-year follow-up case. AB - We report a patient with unusual glomerulonephritis. A 24-year-old Japanese female was hospitalized in October 1995 because of nephrotic syndrome. Lobular form glomerulonephritis with mesangial proliferation associated with massive wide spread accumulation of slightly eosinophilic, periodic acid Schiff-positive amorphous materials in the luminal side of the capillary walls and paramesangial area was observed in the renal biopsy specimen. Immunofluorescent study revealed massive strong staining for IgM and C4 along the capillary walls and in the mesangium. Deposits of IgA, IgG, C3 and fibrinogen were also observed. Electron microscopy showed normal thickness of the capillary basement membrane and a large amount of subendothelial and paramesangial electron dense, finely granular deposits without fibrils or tubular structures. There were no clinical or laboratory findings of systemic diseases, such as systemic lupus erythematosus and cryoglobulinemia. Therefore, we believed that this case involved an unusual idiopathic glomerular disease with massive subendothelial and paramesangial immune deposits. Glomerulonephritis in this patient appeared to be resistant to treatment with corticosteroids and that this glomerulopathy may be a progressive disease as shown during the 3-year observation. Furthermore, our patient had idiopathic hyperprolactinemia and subclinical hypothyroidism. However, the relationship between glomerulonephritis and endocrinopathy in our patient is unknown. PMID- 9403219 TI - Systemic lupus erythematosus in a patient with remitting minimal change nephrotic syndrome. AB - Minimal change nephrotic syndrome (MCNS) developed in a 17-year-old female and spontaneously remitted. One month later the nephrotic syndrome relapsed. Prednisolone therapy, 60 mg/day, was started and resulted in a full remission within a week and the prednisolone dose was subsequently tapered. Seven months later, when 10 mg/day of prednisolone was being administered, she developed erythematous rash with photosensitivity and polyarthralgia without exacerbation of the nephrotic syndrome, and fulfilled four of the American College of Rheumatology criteria for classification of systemic lupus erythematosus (SLE). Avoidance of direct sunlight ameliorated the erythematous rash and the polyarthralgia disappeared even though the prednisolone dose was decreased further. This is the first reported case of SLE developed in a patient with remitting MCNS. PMID- 9403220 TI - Acute renal failure with severe tubulointerstitial changes in a patient with minimal change nephrotic syndrome treated with enalapril. AB - A 35-year-old nephrotic man developed acute renal failure with serum creatinine to 1543 micromol/l after a month of therapy with enalapril. Renal biopsy demonstrated minimal glomerular changes with fusion of podocytes, tubular necrosis with regeneration of tubular epithelial cells, interstitial edema with focal interstitial fibrosis, and interstitial infiltration with neutrophils, eosinophils, plasma cells and mononuclear cells. Three hemodialyses were performed in the patient during the oliguric phase of the disease. Renal function was restored after withdrawal of enalapril and initiation of steroid therapy. Steroids also contributed to the improvement of the nephrotic syndrome and proteinuria decreased from maximal ranges of 27 g/l to 2.2 g/l after six months of the follow-up. Similar cases were previously described associated with captopril treatment, but not with enalapril. PMID- 9403221 TI - Losartan for therapy of posttransplant erythrocytosis. PMID- 9403222 TI - Amlodipine has a minor effect on cyclosporine metabolism. PMID- 9403223 TI - Role of peripheral-type benzodiazepine receptors in steroidogenesis. AB - Peripheral-type benzodiazepine (BZ) receptors (PBRs) have been identified in various peripheral tissues as well as in glial cells in in the brain. PBRs are located mainly on the outer mitochondrial membrane and bind with high affinity the BZ Ro 5-4864 (4'-cholorodiazepam) and the non-BZ PK 11195 (an isoquinoline carboxamide derivative), but bind with very low affinity the BZ clonazepam. PBRs have been cloned from various species. PBRs are multimeric receptors composed of the 18-kDa binding site for isoquinolines, the 32-kDa voltage-dependent anion channel, and the 30-kDa adenine nucleotide carrier (which binds BZs). The expression of PBRs is especially high in steroidogenic organs. Steroid administration affects PBR density, whereas depletion of hormones by hypophysectomy in female rats, or castration (surgical or chemical) in male rats, decreases PBR density in endocrine organs, which can be elevated to normal values after administration of the appropriate hormone. PBRs are probably involved in several functions, including cell proliferation, respiration, and steroidogenesis. It has been suggested that PBRs are involved in the translocation of cholesterol from the outer to the inner membrane of the mitochondria and have an effect on the biosynthesis of steroids. PMID- 9403224 TI - Melatonin, its receptors, and relationships with biological rhythm disorders. AB - Melatonin is a neurohormone produced during the night by the pineal gland. Its secretion is regulated by circadian and seasonal variations in daylength, transmitted via visual projections to the suprachiasmatic nucleus which functions as a circadian clock in mammals. Melatonin has been proposed to act as an internal synchronizer of circadian rhythms generated at different levels of the organism. The chronobiotic effects of melatonin in humans have been mainly studied in circadian rhythm sleep disorders related to jet lag, shift work, blindness or aging. Alterations of the melatonin profiles have also been reported in other biological rhythm disorders. PMID- 9403225 TI - Effect of citalopram on brain serotonin release in experimental hepatic encephalopathy: implications for thymoleptic drug safety in liver insufficiency. AB - In the present study, effects of citalopram (CIT) on brain 5-hydroxytryptamine (5 HT) release in experimental chronic hepatic encephalopathy (HE) were investigated. Neocortical administration of CIT (1.0 microM) increased the brain 5-HT output to a similar extent in portacaval shunted (PCS) rats and sham operated controls, indicating that a previous described mismatch between increased 5-HT turnover and unchanged release in PCS rats is not explained by an accelerated brain 5-HT reuptake. Subsequent systemic administration of CIT (5 mg/kg subcutaneously) resulted in a more pronounced attenuation of the brain 5-HT release in PCS rats than in sham-operated controls, possibly indicating a higher susceptibility to indirect mid-brain 5-HT1A autoreceptor activation in experimental portal-systemic encephalopathy (PSE). A KCl (60 mM) challenge in the presence of locally administered CIT (1 microM) induced a more marked neocortical 5-HT response in PCS rats than in sham-operated controls, confirming previous results of a higher than normal amount of 5-HT available for depolarization induced release in PCS rats. Although the pharmacodynamic parameters in this study was investigated for CIT, the likelihood of a parallel pharmacokinetic alteration existing for this drug in the PCS condition also was indicated. It is thus suggested that otherwise generally safe central nervous system 5-HT-active drugs may represent a potential hazard in patients with liver failure with or without PSE. PMID- 9403226 TI - Motor complications of chronic levodopa therapy in Parkinson's disease. AB - We report on motor complications of chronic levodopa therapy among 811 levodopa responsive patients with idiopathic Parkinson's disease (PD), stratified by duration after diagnosis. Predictable "offs" were noted in 20.2% of patients in the first 5 years, in 58.3% after 15 years. Unpredictable or sudden offs and early morning dystonia were less common. Longer duration was associated with greater percentages of patients with off periods or dyskinesias (up to 70% after 15 years), although patients with 6-15 years' duration saw relatively little increase in frequency of those complications, and a minority of patients (approximately 30%) with duration into the second decade did not experience off periods or dyskinesia. Across groups, mean Hoehn and Yahr stage and daily levodopa dosage progressively increase (and mean Schwab and England disability ratings decrease), but more conservatively than in prior reports in the postlevodopa era. We note that with advancing PD duration, levodopa complications are more common, but in many cases there appear to be relatively stable periods in terms of levodopa dosage and disease severity, and a minority of patients will be relatively free of motor complications into the second decade of their disease. PMID- 9403227 TI - Effect of tolcapone on plasma levodopa concentrations after coadministration with levodopa/carbidopa to healthy volunteers. AB - Tolcapone (Ro 40-7592) is a novel inhibitor of catechol-O-methyltransferase that is being developed for clinical use in the treatment of Parkinson's disease as add-on therapy to a combination of levodopa and a peripheral amino acid decarboxylase inhibitor (benserazide or carbidopa). The current single-blind, randomized study was designed to evaluate the effect of tolcapone compared with placebo on plasma levodopa concentrations in healthy volunteers concomitantly receiving 25 mg of carbidopa and 100 mg of levodopa (Sinemet 25-100) and to assess the tolerability and safety of this combination. Placebo or tolcapone at doses of 5, 10, 25, 50, 100, 200, 400, and 800 mg was coadministered orally with Sinemet 25-100. Each dose was tested in a crossover fashion in a new group of six participants who each received active drug on one occasion and placebo on the other. Tolcapone increased the area under the plasma concentration-time curve and half-life of levodopa approximately twofold, without appreciably increasing the peak concentration. The maximum effect on levodopa half-life was observed with the 200-mg dose. Adverse effects were minor at all doses. PMID- 9403228 TI - Selegiline and excess mortality. PMID- 9403229 TI - Coronary intervention in myocardial infarction. AB - The ability to achieve rapid and sustained reperfusion in cases of acute MI with an interventional approach constitutes a true revolution in the practice of cardiology. This article discusses those interventions. PMID- 9403230 TI - Colon rectal cancer: a common disease. AB - Early diagnosis is a key in properly screening and surveillance. The author discusses preventive medicine, risk ratings, and fecal occult blood testing. PMID- 9403231 TI - Lung transplantation: an overview. PMID- 9403233 TI - Should circulating anticoagulant [AKA anti-phospholipid] assays be a routine component of well-patient assessment? AB - Circulating anticoagulants are a major risk factor for thrombotic problems (eg, myocardial infarction, stroke) and pregnancy complications. The authors present a retrospective survey of anticardiolipin antibody and lupus anticoagulant in 200 consecutive patients presenting to their office. PMID- 9403232 TI - New developments in osteoporosis. AB - Osteoporosis is no longer a disorder about which nothing can be done. Many of the new developments in this field have already assumed important places in clinical management. The author reviews many of the new developments in treating osteoporosis and attempts to place them in a practical, clinical context. PMID- 9403234 TI - Self-directed work teams: an 18-month review. AB - Visionaries are individuals who imagine how the ideal practice should be setup. Realists view practices the way they are actually setup. To bring these individuals together toward a common goal is what self-directed work teams can provide your practice. Components that make up a self-directed work team are reviewed. PMID- 9403235 TI - Benefit plans in a changing practice environment. AB - Managed care is affecting the business of medical practices. Many physicians are joining with larger organizations and employee benefits are even more significant in each physician's financial security. Background on benefit issues and common areas of caution when contemplating joining a large entity are addressed. PMID- 9403236 TI - Cerebral evoked responses to gastrointestinal stimulation in humans. AB - Recent advances have permitted recording of evoked potentials (EPs) in response to electrical and mechanical stimulation of the gastrointestinal (GI) organs via methods used primarily in clinical neurophysiology. Current research involving stimulation of the esophagus, rectum, and colon, and recording the corresponding responses on the scalp, is being practiced in only a few laboratories. This review examines the engineering aspects of recording EPs, such as characteristics of the stimuli, placement of stimulus electrodes in the GI tract, and enhancement of evoked potential signals. We also discuss the physiological concepts involved in the generation of EPs, and how these compare with somatosensory evoked responses. Current experimental techniques employed by various investigators and results reported from their laboratories are compared. We believe that cerebral EPs to GI stimulation could be useful in studying a number of pathophysiological conditions such as gastroesophageal reflux disease, diffuse esophageal spasm, chronic inflammatory bowel disorders, chronic abdominal pain, and irritable bowel syndrome, among others. We hope that the present review will generate interest in the use of EPs arising out of GI stimulation, aiding in understanding their physiological implications in healthy subjects and in GI disorders. PMID- 9403237 TI - Biomechanics of atherosclerotic plaque. AB - Atherosclerosis is the leading cause of death in the U.S. In balloon angioplasty, pressure is applied directly to atherosclerotic plaque to reopen the occluded blood vessel. The mechanical behavior of the plaque often determines the outcome of the angioplasty. Little information on the material properties of atherosclerotic plaque is available, yet the properties govern the plaque's behavior. Our discussion of the experimental testing and numerical analysis of plaque is directed toward summarizing the current knowledge of plaque material properties. Atherosclerotic plaque exhibits a wide range of behaviors consistent with the variability in the underlying composition. Overall, plaques exhibit nonlinear and inelastic mechanical behavior, although geometry and material properties are not well known. The histomorphological composition is critical in determining the plaque's mechanical response. Finite element approximations have been used to study the stresses developed in the diseased vessel; however, material properties are a critical component of a finite element analysis: the predictive capabilities depend on how accurately the material is modeled. When more information on plaque behavior is generated through careful and extensive experimental investigations, better models will be constructed to more accurately predict plaque responses. As the biomechanics community learns about plaque mechanics, we can use the knowledge to enhance the reliability of interventional procedures. PMID- 9403238 TI - Dental mercury amalgam: Part II. Safety of mercury amalgam. PMID- 9403239 TI - Aural vignettes of dermatologic history. Interview by Victor H Witten. PMID- 9403240 TI - Verrucous plaque on the back of a hand. AB - We report the case of a 44-year-old slaughter-house operator with a skin lesion that had been present for two years on the back of his left hand. The lesion had increased progressively in size despite numerous topical treatments. Physical examination revealed an infiltrated erythematous-violet plaque with a verrucous surface featuring numerous orifices draining purulent material. Histologic study of the lesion disclosed tuberculoid granulomatous infiltrates at the dermoepidermal limit. Presence of Mycobacterium tuberculosis, together with other epidemiologic, clinical, histologic, and immunologic data, permitted a diagnosis of tuberculosis verrucosa cutis to be made. The excellent response of the patient to treatment confirmed this hypothesis. However, polychemotherapy withdrawal was temporarily needed due to analytical abnormalities. PMID- 9403241 TI - Recurrent condyloma acuminatum due to piercing of the penis. AB - An unusual case of recurrent condyloma acuminatum in a tubular penile piercing scar is presented. This case documents a previously unreported dermatologic infectious sequela of this practice. We comment on complications of bodily and genital piercing. PMID- 9403242 TI - Vitiligo. AB - Vitiligo is a common disorder afflicting people of all ages around the world. It produces milky white patches of depigmentation that cause significant morbidity due to cosmetic disfigurement. One should distinguish segmental from nonsegmental vitiligo, and be aware the latter is associated with an extremely small risk of associated autoimmune disorders. New therapeutic options offer some additional hope to patients with vitiligo, but more progress in the understanding of its pathogenesis and therapy is still needed. PMID- 9403243 TI - Linear focal elastosis associated with striae distensae in an elderly woman. AB - We report the case of a 73-year-old woman with linear focal elastosis (LFE) associated with striae distensae. Yellow palpable striae were found extending horizontally in the lumbar region. The yellow striae appeared to be joined to striae distensae in the lateral sides of the yellow striae. Striae distensae were also present in regions other than the back but were not associated with yellow striae. Histologic examination of a yellow stria revealed a focal increase in elastic fibers in the dermis. Although there have been some unusual cases of LFE arising in women or in young persons, this disorder still predominantly affects aged men. A fairly uniform feature of LFE is its occurrence on the dorsal skin, suggesting involvement of this specific anatomical site in its pathogenesis. We suspect that LFE usually arises de novo in the skin, although striae distensae may also be a cause of it. PMID- 9403244 TI - Hair casts: a case of pseudonits. AB - Hair casts are often misdiagnosed because of their close resemblance to nits from an infestation with pediculosis capitis. In their clinical presentation both of these disorders may appear to have white keratinous material adherent to hair shafts and can look very similar. Microscopic examination provides the definitive and correct diagnosis. We report a case of hair casts, also referred to as pseudonits. PMID- 9403245 TI - Trichophyton rubrum onychomycosis in a 17-year-old girl. AB - Onychomycosis is one of the most common nail disorders seen by dermatologists. An increasing number of immunocompromised patients, in combination with an aging worldwide population, has resulted in a significant increase in the incidence of this disorder. While nearly one fifth of all occurrences are found in patients aged 40 to 60, the incidence dramatically increases in patients over 60. However, onychomycosis is very uncommon in children. PMID- 9403246 TI - Massive dystrophic calcinosis cutis secondary to chronic needle trauma. AB - Extensive dystrophic calcinosis cutis of the hips and anterolateral thighs in a woman with insulin-dependent diabetes mellitus is described. Clinical, histologic, and radiographic data are provided to familiarize the reader with this unique and newly described condition. We speculate that repetitive, chronic, needle trauma in susceptible persons can induce massive dystrophic calcinosis cutis. PMID- 9403247 TI - Leukemia cutis as the initial manifestation of acute nonlymphocytic leukemia in a young child. AB - A 15-month-old boy was well except for asymptomatic, erythematous, wheal-like papuloplaques, macules, and nodules on his face and four extremities. It was misdiagnosed by a pediatrician and treated as urticaria for six months. Later, he was sent to our hospital for evaluation of prolonged fever. Acute nonlymphocytic leukemia (M5) with leukemia cutis was diagnosed by results of hematologic examination and examination of a skin biopsy specimen. After one course of chemotherapy, all of the skin lesions completely resolved and had not recurred. Five months after acute nonlymphocytic leukemia was diagnosed, bone marrow relapse and central nervous system involvement were noted. PMID- 9403248 TI - Povidone-iodine in antisepsis--state of the art. AB - The natural element iodine has been used for more than 150 years to prevent infection and treat wounds. Yet only due to the development of iodophors has it become possible to use this highly efficient microbicide in a wide range of medical applications. The antimicrobial spectrum is universal. Its efficiency against clinically and epidemiologically significant new pathogens, such as methicillin-resistant Staphylococcus aureus and Enterococcus sp. has also been validated. No development of resistance has been determined. New data are also available on the excellent local tolerability of Betaisodona (povidone-iodine) preparations. On these grounds, a number of clinical fields exist in prophylaxis and therapy, for either once only or repeated applications: the disinfection of hands and skin, mucosa antisepsis, intra- and postoperative wound treatment, therapy of skin infections, burns and chronic wounds. PMID- 9403249 TI - Nondevelopment of resistance by bacteria during hospital use of povidone-iodine. AB - Since the bacterial ability to develop resistance against various factors of their surroundings is a well-known phenomenon, resistance against iodine and specifically against povidone-iodine (PVP-I) has been widely investigated. Yet there is little known about bacterial resistance in long-term daily use of disinfectants in continuous ambulatory peritoneal dialysis (CAPD) patients. The aim of our study was to investigate whether on daily use of PVP-I over a period of at least 6 months coagulase-negative staphylococci (CNS)--the predominant infective organisms of peritonitis--developed resistance against PVP-I. At the catheter exit site of 40 CAPD patients we isolated 36 CNS. 23 CNS (CNS + PVP) orginate from patients using PVP-I, 13 CNS (CNS + CI) from patients using sodium hypochlorite (NaOCl) as disinfectant. The strains were biotyped, antibiotic resistance patterns were determined and resistance against PVP-I or NaOCl was calculated as reduction factor using the quantitative suspension test combined with a turbidimetric standardization. Resistance against PVP-I 0.01% and against NaOCl 0.005% was determined at two contact times (30 and 300 s) for each patient group. In addition, we investigated the effects of plasmid loss on sensitivity to PVP-I. Out of 5 multiple-antibiotic-resistant CNS, 3 strains showed no difference in reduction factor against PVP-I before and after curing. There was no significant difference in reduction factor against NaOCl. CNS + PVP were even significantly more sensitive to PVP-I than CNS + Cl. Taken together, our results demonstrate that long-term use of PVP-I does not cause any bacterial resistance in CNS of CAPD patients. PMID- 9403250 TI - Investigation on the efficacy of povidone-iodine against antiseptic-resistant species. AB - The bactericidal activity of commonly used antiseptics against clinical isolates was determined. It is noteworthy that povidone-iodine (PVP-I) solution showed high bactericidal activity against all of the test strains after 30 s of exposure. However, in the case of chlorhexidine gluconate (CHG), residual bacteria were observed in most species. Next, acquisition of resistance to antiseptics was examined, and it was found that remarkable increases in MICs were seen for CHG and alkyldiaminoethylglycine hydrochloride. The strains which acquired resistance against one antiseptic showed cross-resistance to all antiseptics except for PVP-I. As for bactericidal activity against biofilm, no viable cells were seen after a 10-min exposure to PVP-I solution. No decrease in viable cell count was seen even after a 60-min exposure to any of the other antiseptics. PVP-I showed high activities in all the tests conducted in this study. Thus, PVP-I was confirmed to be a clinically useful antiseptic. PMID- 9403251 TI - Comparison of bactericidal effects of commonly used antiseptics against pathogens causing nosocomial infections. Part 2. AB - Opportunistic infections caused by gram-negative rods (GNR), conventionally regarded as organisms with low or no pathogenicity, and intractable infections caused by various resistant organisms pose a great problem now. In view of this, we determined the bactericidal effects of 5 commonly used disinfectants using as the test strains Xanthomonas maltophilia and Serratia marcescens, chosen among other GNR since they often cause nosocomial infections. Regarding the bactericidal activities against X. maltophilia and S. marcescens, both sensitive strains and resistant strains were killed within 20 s of exposure to povidone iodine and sodium hypochlorite. With chlorhexidine, 1 strain each of both species was not killed within 10 min of exposure at a concentration of 0.2%. Both sensitive strains and resistant strains of X. maltophilia were killed within 20 s of exposure to benzalkonium at 0.02%, while a concentration of 0.1% was required for benzalkonium to kill S. marcescens within 20 s. With Tego-51, both sensitive strains and resistant strains of X. maltophilia were killed within 20 s at 0.02%, while 1 strain of S. marcescens was not killed within 20 s at a concentration of 0.1%. In the use of disinfectants, comparative bactericidal effects of various disinfectants against clinical isolates should be taken into consideration. PMID- 9403252 TI - Inactivation of human viruses by povidone-iodine in comparison with other antiseptics. AB - Inactivation of a range of viruses, such as adeno-, mumps, rota-, polio- (types 1 and 3), coxsackie-, rhino-, herpes simplex, rubella, measles, influenza and human immunodeficiency viruses, by povidone-iodine (PVP-I) and other commercially available antiseptics in Japan was studied in accordance with the standardized protocol in vitro. In these experiments, antiseptics such as PVP-I solution, PVP I gargle, PVP-I cream, chlorhexidine gluconate, alkyldiaminoethyl-glycine hydrochloride, benzalkonium chloride (BAC) and benzethonium chloride (BEC) were used. PVP-I was effective against all the virus species tested. PVP-I drug products, which were examined in these experiments, inactivated all the viruses within a short period of time. Rubella, measles, mumps viruses and HIV were sensitive to all of the antiseptics, and rotavirus was inactivated by BAC and BEC, while adeno-, polio- and rhinoviruses did not respond to the other antiseptics. PVP-I had a wider virucidal spectrum, covering both enveloped and nonenveloped viruses, than the other commercially available antiseptics. PMID- 9403253 TI - Antiseptic efficacy of disinfecting solutions in suspension test in vitro against methicillin-resistant Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli in pressure sore wounds after spinal cord injury. AB - In pressure sore wounds after spinal cord injury, methicillin-resistant Staphylococcus aureus can be detected in 2% of the cases. The elimination of the germ is the aim of the treatment. Pressure sore wounds are an often found complication after spinal cord injury. For local treatment five commercially available antiseptics for the skin and mucous membrane were tested in vitro. The method used is a modified qualitative and quantitative suspension test. The antiseptics were tested without and with addition of 5% albumin in order to simulate the conditions of the wound in vivo. The results show a superior efficacy of the povidone-iodine preparations. Betadine, probably due to the higher concentration, is more efficacious than Braunol; chlorhexidine is sufficiently efficacious without the addition of albumin. These results still have to be confirmed by in vivo studies. PMID- 9403255 TI - Profile of patients treated with Betadine cream and ointment in a major burn centre over a period of ten years (1986-1995). AB - The management of burns has shown substantial progress over the years. The application of an effective topical agent improves the healing rate of burns. Betadine cream and ointment have been used in all the burn admissions since 1986. The data collected over 10 years are analysed in this report. All the burn patients admitted were evaluated and the factors investigated included patient age, sex, cause of injury, extent of burn wound, length of hospital stay, bacteriology, multi-organ failure and mortality. A total of 6,056 patients were admitted to a 40-bed unit during the 10-year period. Sixty-two percent were adults and 37% children. There were 67.5% males and 32.5% females. Eighty-one percent were admitted directly to the unit. Intensive/high care admissions accounted for 19.2% patients. The major cause of injury was accidental scalding in 34.2% patients, and the majority (65.4%) stayed between 1 and 4 weeks in hospital. There were 452 deaths. This audit reviewing 6,056 sequential burn cases represents a 10-year experience of burn management, using Betadine cream and ointment as topical agent. PMID- 9403254 TI - Effects of Betaisodona on parameters of host defense. AB - The numbers of patients in intensive care units, with immunosuppression, and of elderly people increase in parallel with antibiotic-resistant microorganisms. Therefore the demand for an effective antisepsis increases. Moreover, it became evident that the pathophysiology and the outcome of infection are dependent on the properties of the microorganisms, e.g. synthesis of endo- and exotoxins, and on the host defense, the immune system. In addition to the microbicidal action, we studied the effects of povidone-iodine (PVP-I, Betaisodona) on the generation, release and activity of exotoxins (alpha-hemolysin, phospholipase C, lipase), as well as on granulocyte-derived tissue-destructive enzymes (elastase, beta glucuronidase) and microbial-induced cytokine generation from human neutrophils. Our results clearly show that PVP-I does not only kill a wide range of bacteria but also inhibits the generation and release of bacterial exotoxins; furthermore, it also inactivates bacterial exotoxins as well as granulocyte-derived tissue destructive enzymes and cytokines. These data support the usefulness and efficacy of PVP-I as an effective therapeutic agent to combat infection. PMID- 9403256 TI - Vaginal antisepsis for hysterectomy: a review of the literature. AB - Infectious complications of hysterectomy remain common despite the use of prophylactic antibiotics. Most are caused by contamination of the surgical site by vaginal bacteria which are not controlled by current methods of pre-operative antisepsis. The medical literature concerning antiseptic preparation of the vagina for surgery was reviewed to discover the evidence on which practice may be based. A search using Medline, Current Contents, the Cochrane Library and the reference lists of articles on the subject and of major gynaecology textbooks produced 13 comparative studies. No conclusive randomized controlled trials were found and most of the studies had severe methodological problems limiting interpretation of their results. The scant available data suggest that use of vaginal antiseptics before the patient arrives in the operating room is probably not useful, and that application of povidone-iodine vaginal gel at the beginning of abdominal hysterectomy is sufficiently promising to justify further investigation. PMID- 9403257 TI - Povidone-iodine to prevent mucositis in patients during antineoplastic radiochemotherapy. AB - In an open study, the efficacy of povidone-iodine in the prophylaxis of mucositis during antineoplastic radiochemotherapy was determined. 40 patients were randomly assigned to a treatment or control group (each 20 patients). All patients received standard prophylaxis of mucositis with nystatin, dexpanthenol, rutoside and immunoglobulin. In addition, the patients of the treatment group performed 4 times daily rinsing with povidone-iodine, the control patients with sterile water. Clinical examination of the oral mucosa was performed weekly during the radiation period and up to 6 weeks after the end of therapy. Oral mucositis was observed in 14 patients of the treatment group (mean grading: 1.0) and in all 20 patients of the control group (mean grading: 3.0). The mean onset of mucositis was after 2.25 weeks in treatment patients and 1.5 weeks in control patients. The mean total duration of mucositis was 2.75 weeks in treatment patients and 9.25 weeks in control patients. The mean AUC values were 2.5 in treatment patients and 15.75 in control patients. All findings were statistically significantly different between the two groups. It is concluded that rinsing with povidone iodine reduces the incidence, severity and duration of oral mucositis during antineoplastic radiochemotherapy. PMID- 9403258 TI - New successful treatment with disinfectant for atopic dermatitis. AB - For the treatment of atopic dermatitis, a variety of therapies are used including folk medicine. At present, there is no single treatment which is effective to cure the symptoms of atopic dermatitis completely in all patients. We are drawing attention to the high isolation rate of Staphylococcus aureus when starting disinfectant treatment combined with topical steroid therapies for the purpose of killing S. aureus. As a result, we examined many patients in whom almost a complete remission was obtained even after short periods of therapy, though it had been difficult to obtain improvement by conventional treatments. In many patients, IgE values and reagin antibody titer decrease dramatically soon after starting treatment. As a disinfectant, 10% povidone-iodine solution was used. We investigated also the effect of iodine contained in the povidone-iodine solution on the thyroid gland. PMID- 9403260 TI - Prevention of catheter-associated urinary tract infection by meatal disinfection. AB - The incidence of catheter-associated urinary tract infections (UTIs) becomes higher with prolongation of the indwelling period of a catheter. As to the entry of bacteria, ascending UTIs have now attracted attention. In the present study. the metal area was examined bacteriologically and the possibility to use antiseptics for blocking the route of developing infections was investigated. The subjects included 72 patients with an indwelling, urethral catheter inserted post operatively. These patients were divided into three groups treated with once or twice daily application of povidone-iodine or once daily application of povidone iodine cream. In these groups, the relation between changes in isolation of bacteria from the meatal area and the incidence of UTI was evaluated. It was found that reduction in bacterial count by antisepsis is effective to prevent ascending UTIs. Moreover, once daily application of povidone-iodine was proven to be effective in male patients. The effective antisepsis in females was twice daily application of povidone-iodine. PMID- 9403259 TI - The influence of long-term treatment with povidone-iodine on thyroid function. AB - Povidone-iodine (PVP-I) is generally very safe, but cases of thyroid dysfunction induced by PVP-I have been reported. The effect of long-term treatment with PVP-I on thyroid function was to be assessed. In 40 inpatients of the department of neurology, the status of the use of PVP-I preparations and their effects on serum inorganic iodine levels and thyroid functions were investigated. In 27 patients treated with PVP-I for a long term, inorganic iodine levels were significantly increased as compared to those in 13 patients without PVP-I treatment. Out of 27 patients treated with PVP-I in the long term, subclinical hypothroidism was seen in 3 patients, mild hyperthroidism was seen in 1 patient, and subclinical hyperthyroidism was suspected in 7 patients. Patients treated with PVP-I for a long time should be observed carefully for any manifestation of thyroid dysfunction. PMID- 9403262 TI - An open-label study conducted to evaluate the efficacy of Betadine cold sore paint. AB - An open-label, randomised, parallel-group efficacy study was designed to compare Betadine cold sore paint and Stoxil topical ointment for the prevention of shedding of herpes simplex virus (HSV) from cold sores. Seventy-two patients aged 18-61 years (mean 32.2 years), with symptoms indicating recent onset of herpes labialis, were entered into the study. Patients were randomised to receive Betadine cold sore paint, Stoxil topical ointment or no treatment. To detect infectious virus, swabs were taken for virus culture before and 2 h after treatment. The no-treatment control group was included to monitor the efficiency of the swabbing technique. The primary measure of efficacy was the proportion of patients in each group returning a swab positive for HSV prior to treatment application and negative for HSV 2 h after treatment application. All 72 patients completed the study. HSV clearance rates were 0.636 for the Betadine treatment group and 0.092 for the Stoxil treatment group (p = 0.00056). It was concluded that recovery of infectious HSV from the lips of patients who applied Betadine cold sore paint to their lesions was significantly lower than from patients who applied Stoxil topical ointment. PMID- 9403261 TI - Antiseptic effect of povidone-iodine solution on abdominal skin during surgery and on thyroid-gland-related substances. AB - No definite guidelines have been published concerning the suitable exposure time and frequency of skin antisepsis of the operative field. In the present study, the antiseptic effect of a single use (single-application group) of 10% povidoneiodine solution for skin antisepsis was compared with its triple use (triple-application group). The exposure time was 2 min for both groups. Moreover, the effects on blood levels of total iodine and thyroid-gland-related substances were evaluated. High antiseptic efficacy was obtained in both groups, indicating that our antiseptic method to disinfect the operative field is effective. Slightly better results were obtained from the triple-application group, although the difference was not statistically significant. Blood levels of total iodine and thyroid-gland-related substances remained within the normal range in all cases. However, in cases receiving a large amount of 10% povidone iodine solution a transient elevation in blood total iodine was observed. Thus, the 10% povidone-iodine solution is considered to be safe and effective for skin antisepsis of the operative field when applied repeatedly in a small amount per dose with an exposure time of about 2 min. PMID- 9403263 TI - Cytotoxicity and sensitization of povidone-iodine and other frequently used anti infective agents. AB - Povidone-iodine is an antiseptic widely used in dermatology. In vitro experiments showed a certain cytotoxicity, yet it is not easy to transfer these toxicological data to in vivo circumstances. In vivo investigations in animals and in humans could exclude cytotoxic effects of povidone-iodine, measured by the wound healing process. Only when administered in combination with detergents was an obvious cytotoxicity seen in wounds but not on the intact skin. In comparison to the frequently used antibiotic neomycin, the sensitization rate of povidone-iodine is very low. PMID- 9403265 TI - Pharmaceutical and bacteriological study on povidone-iodine sugar ointment. AB - Povidone-iodine sugar ointment is an excellent preparation for the treatment of decubitus. It has been used as an intrahospital preparation made according to the formula each hospital decided on from experience. Although commercial products have also been developed and used, they are too expensive. The efficacy of a povidone-iodine sugar ointment formulation which can be prepared by a single method and which has the stability and antibacterial activity equal to commercially available products was evaluated. As the test drugs, one commercially available product (UP), and three preparations with different formulas (P-1, P-2 and P-3) were used. All of these test drugs were stored at 20 and 40 degrees C. Specimens were sampled immediately after storage and after 20, 60, 90, 120 and 150 days and examined pharmaceutically (measurement of pH value and determinations of available iodine and sucrose levels). For the determination of bacteriological effects, 5 standard strains of 5 genera and 5 strains of methicillin-resistant Staphylococcus aureus (MRSA) were used and the time required to kill the bacteria was determined. For UP and P-3, no changes were seen pharmaceutically after 150 days of storage at 20 and 40 degrees C. However, MRSA could not be killed within 30 min. P-1 and P-2 showed remarkable changes pharmaceutically after 60 days of storage at 40 degrees C and could not be used any more. It became possible to make a preparation of povidone-iodine sugar ointment which has a stability almost similar to that of UP. Moreover, such a preparation can be made at low cost. However, since the bactericidal activity against MRSA was not higher than those of other drugs, the future task is to improve the bactericidal activity. PMID- 9403264 TI - Povidone-iodine liposomes--an overview. AB - In recent years, liposomes have been increasingly explored as novel drug delivery systems, and several liposome-based drug products have been approved in Europe, the USA and Japan. Depending on size, composition and surface characteristics, liposomes interact specifically with biological structures. Liposomal drug products provide a topical activity at the desired locus of action and are deemed more effective and less toxic than conventional drug formulations. The combination of povidone-iodine (PVP-I) and liposomes unites the exceptional microbicidal activity of the antiseptic substance with the excellent tolerability and lack of immunogenicity of liposomes; in addition, liposomes provide a moist molecular film for the wound environment. The multilamellar vesicles act as microreservoirs hence prolonging the release of the active ingredient. Although no commercial product for repeated application on the eye is currently available, PVP-I has been used in ophthalmology not only for pre- and postoperative antisepsis, but also for the treatment of bacterial and viral conjunctivitis and for prophylaxis against ophthalmia neonatorum. For these indications, liposomal formulations with 2.5 and 5.0% PVP-I were developed. These eye drops are isotonic with tear fluid at pH 6. First in vitro tests demonstrated an excellent antimicrobial efficacy, and a placebo-controlled clinical study on volunteers showed a very good local tolerability. A study on rabbits demonstrated positive results of the PVP-I liposome eye drops compared to placebo and the broadspectrum antibiotic Polyspectran in a standardized model of Staphylococcus aureus deep eye infection. The other aim is a well-tolerated liposomal PVP-I hydrogel for improved antiseptic wound treatment with moisturizer. It has been reported that liposomes are enriched at the wound bottom for direct action against infection and support of wound healing. An animal study on the efficacy and tolerability of different formulations of a hydrogel with PVP-I liposomes in deep dermal burn wounds has indicated an outstanding quality of wound healing with smooth granulation tissue, less inflammation, less wound contraction and no hyperkeratotic reactivity, especially with the 3% PVP-I liposome formulation. PMID- 9403266 TI - Bactericidal activities of povidone-iodine against Mycobacterium. AB - Three standard strains of Mycobacterium (M. tuberculosis H37Rv, M. avium ATCC15769 and M. kansasii ATCC12478) and 15 clinical isolates of Mycobacterium (7 M. tuberculosis, 2 M. avium, 3 M. kansasii, 1 M. intracellulare, 1 M. chelonae subsp. abscessus and 1 M. gordonae) were selected in order to study the bactericidal activities of povidone-iodine (PVP-I) drug substance and a commercially available PVP-I solution (Isodine solution) against Mycobacterium. After the bacilli had been exposed to the disinfectant solution at concentrations of 0.1 or 0.02% with 2% human serum for various incubation periods from 30 to 120 s, the PVP-I drug substance was inactivated by addition of 0.5% sodium thiosulfate. In the case of the commercially available PVP-I solution, a mixture of 10% Tween 80, 3% soybean lecithin and 0.5% sodium thiosulfate was used as inactivator. It was demonstrated that the 3 standard strains were completely inactivated within 30 s by 0.1% PVP-I drug substance and that the 15 clinical isolates were almost killed by 0.1% commercially available PVP-I solution within 60 s. As a result, the commercially available PVP-I product appeared to be a useful agent as disinfectant against all the tested species of Mycobacterium. PMID- 9403267 TI - Antiseptic effects at injection sites. AB - To our knowledge, there are no published papers detailing antisepsis for injection sites. In view of this, the efficacies of povidone-iodine (PVP-I) ethanol solution and chlorhexidine (CH) ethanol, the agents most commonly used for antisepsis of the operative field, were compared. Before and after the injection site was disinfected with either of these antiseptics, specimens of indigenous bacteria on the skin were collected by the cylinder scrub method, and the bacteria reduction rate and the reduction factor (RF) were determined to evaluate the efficacy of antisepsis. The bacteria reduction rate and RF value obtained for PVP-I ethanol were 95.1 +/- 11.2 and 2.1 +/- 0.9% and those for CH ethanol were 93.5 +/- 9.3 and 1.8 +/- 0.9%. Since there were individual differences in cell count before antisepsis, no significant difference was seen in bactericidal activity. However, slightly more favorable results were obtained with PVP-I ethanol. Although it is impossible to eradicate completely the indigenous microbes with currently available methods, it is considered important for the prevention of infection of the injection site to decrease bacterial counts as much as possible. PMID- 9403268 TI - Molecular effects of povidone-iodine on relevant microorganisms: an electron microscopic and biochemical study. AB - The aim of this study was to elucidate the effects of povidone-iodine (PVP-I) on cell ultrastructure by electron microscopy and to monitor changes in enzyme activity and nucleotide efflux. Staphylococcus aureus, Escherichia coli and Candida albicans, medically relevant gram-positive, gram-negative and yeast microorganisms, served as models. In the presence of PVP-I, rapid partitioning of the cytoplasm and pronounced coagulation of nuclear material was noted. E. coli and S. aureus showed no major structural wall damage. C. albicans exhibited a rapid, dose-dependent 'loosening' of the cell wall; cells remained intact without lysis, rupture or wall breakage. Changes in beta-galactosidase and nucleotide concentrations were measured in E. coli. A rapid and dose-dependent loss of cellular beta-galactosidase activity was found, with no increase in the supernatant; loss of cellular nucleotides corresponded with an increase in the supernatant. Electron-microscopic and biochemical observations support the conclusion that PVP-I interacts with cell walls of microorganisms causing pore formation or generating solid-liquid interfaces at the lipid membrane level which lead to loss of cytosol material, in addition to enzyme denaturation. PMID- 9403269 TI - Antiproliferation effects of hexadecylphosphocholine on solid tumour and leukaemia selectively in vitro. AB - Hexadecylphosphocholine (HePC), an alkylphospholipid analogue representing a new class of antitumor agents, exerts sufficient oncolytic potencies that it might be developed as a selective antitumour drug. This paper reports that 12 cell lines of leukaemia and solid tumour employed in this study were inhibited by HePC in vitro. This result indicated that HePC possessed general antitumour properties, but did not interfere significantly with cell proliferation of normal BMC in tests of colony formation in granulocytic-macrophage colonies (CFU-GM). Because of these advantages, we suggest that HePC would be an excellent antitumour agent for selectively killing tumour cells and is of low toxicity as compared to conventional cytotoxic agents. PMID- 9403270 TI - Effect of heparin on cell proliferation: lack of correlation with heparin binding sites on cell membrane. AB - In this study an attempt was made to correlate the in-vitro anti-proliferative effect of heparin with the heparin binding on the cell surface. Cells with different sensitivities to the anti-proliferative effect of heparin (BHK-21, FAO, SMC, BAEC, A-431, V-79, and skin fibroblasts) were incubated with [3H]heparin either in the presence or in the absence of unlabelled heparin. A saturable binding was found only in BHK-21, FAO, SMC, BAEC and V-79. Scatchard analysis revealed the presence of a single class of binding sites. The binding of [3H] heparin was efficiently displaced by unlabelled heparin, pentosan polysulfate and low-molecular-weight heparin, but not by dermatan sulfate. Although the sensitivity to the anti-proliferative effect of heparin varied considerably among the cell types (BHK-21 > SMC, FAO > BAEC > V-79), there was no correlation between the reduction of proliferation of these cells and either their heparin binding capacity or the number of binding sites per cell. PMID- 9403271 TI - Effect of peroxovanadate compound on phosphoenolpyruvate carboxykinase gene expression and lipid metabolism in diabetic rats. AB - Although it was proved that peroxovanadate-nicotinic acid (POV) can decrease hyperglycaemia and hyperlipaemia, its molecular biochemical mechanism of action is unclear. The present investigation aimed at studying the effect of POV on gene expression and enzymatic activity of hepatic phosphoenolpyruvate carboxykinase (PEPCK) and blood lipid metabolism in streptozotocin-diabetic rats in order to suggest the molecular biochemical mechanism of POV action in lowering hyperglycaemia and hyperlipaemia. The results showed that the gene expression and enzymatic activity of hepatic cytosolic PEPCK, which were increased in diabetic rats, were significantly reduced following POV treatment. Similarly, POV was shown to oppose significantly the hyperglycaemia and hyperlipaemia in these diabetic rats. These results suggested that a possible mechanism of POV action was to inhibit PEPCK gene expression as well as PEPCK activity which could explain the reduced gluconeogenesis and hyperglycaemia. PMID- 9403272 TI - Effect of acute and chronic inflammation on plasma growth hormone levels in rats. AB - We investigated the effect of acute and chronic stress on growth hormone (GH) plasma levels in rats. Acute stress was provoked by intravenous administrations of IL-1 beta and TNF-alpha. Determinations were made at 10, 30, 60, 120 and 180 min following i.v. injection of these cytokines into the caudal vein. We also investigated the chronic stress induced by hind paw injections of Freund's adjuvant. Arthritis was developed by 21 days following such injection. GH levels were studied at 7, 14 and 21 days after induction of arthritis on several blood samples which were withdrawn from tail veins, and long-term hormonal profiles (3 hours' sampling) were determined at 12.00 am, 1.30 pm and 3.00 pm. Local administration of dexamethasone and the monoclonal antibody anti-ICAM-1 were also used in arthritic rats. Following acute stress, a significant reduction of plasma GH levels has been evidenced, possibly related to the stimulation of corticotropin-releasing hormone. Following chronic stress, we demonstrated a significant increase of GH levels, which were significantly reduced by dexamethasone treatment and to a lesser extent by anti-ICAM-1 administration. PMID- 9403273 TI - Paradoxical effects of intra-arterial administration of basic fibroblast growth factor on inflammatory angiogenesis in rats. AB - The authors studied the effects of basic fibroblast growth factor (b-FGF) on inflammatory angiogenesis in rats. In the corneal cauterization model b-FGF was given intra-arterially (i.a.) (carotid) and in the mesenteric window angiogenesis model, topically (i.e., intraperitoneally (i.p.)). The corneal cauterization was done under anaesthesia by topical application of silver nitrate. Mesenteric window angiogenesis was induced by injection of saline or b-FGF for four days. There were the same two groups of treatment in both models b-FGF 2.5 micrograms/kg/day or saline 1.2-5 ml/kg/day. The area of neovessels and the number of polymorphonuclear cells/field were considered for the corneal angiogenesis, the total length of neovessels was measured for the mesenteric window angiogenesis. The results were expressed as mean values (s.d.). When given i.a., b-FGF significantly reduced the number of polymorphonuclear cells three days after corneal cauterization (from 107 +/- 27 to 41.8 +/- 26, p < 0.01) and inhibited the area covered by neovessels (30 +/- 7.7% vs 51 +/- 20%, p < 0.01) after five days. In contrast, given through the extracellular space, it significantly stimulated the length of mesenteric window microvessels (169 +/- 60 mm vs 90 +/- 31 mm, p < 0.05). These results suggest that b-FGF stimulates inflammatory angiogenesis through interaction with extracellular matrix components, but inhibits it directly when given intra-arterially. PMID- 9403274 TI - Phase I clinical trial of cefditoren pivoxil (ME1207). Tolerance in healthy volunteers. AB - A tolerance study to ME1207 was conducted in healthy volunteers using five single doses administered before a meal (50, 100, 200, 300 and 500 mg), a single 200 mg dose administered after a meal, and 200 mg administered every 12 h for 7 days. No subjective symptoms or changes in haematological and biochemical examinations and urinalysis were observed after single administration of 50 to 500 mg Four subjects complained of minimal upper abdominal discomfort when 200 mg was administered every 12 h for 7 days, but this symptom disappeared without remedial action in all cases. Therefore ME1207 was thought to be safe when orally administered at the above-mentioned doses by the above-mentioned administration methods. PMID- 9403275 TI - Possible involvement of nitric oxide in the quinolone-induced tendon lesions in rats. AB - Quinolone antibacterial agents have been reported to induce adverse effects on the tendon and the musculoskeletal system in humans. We have previously demonstrated that Achilles tendon lesions could be induced in juvenile rats by a single oral administration of quinolones at high doses with simultaneous induction of lesions in the muscle, synovial membrane and articular cartilage. In the present investigation, we examined the involvement of nitric oxide (NO) in pefloxacin (PFLX)-induced lesions of the Achilles tendon in juvenile rats. The incidence of lesions was diminished markedly by co-administration of a potent NO synthase inhibitor, N-nitro-L-arginine methyl ester (L-NAME). Further, the urinary nitrate/nitrite excretion was decreased significantly by 4 h after administration, unchanged between 4 and 8 h, and significantly increased in the 8 to 24-h samples in the PFLX group, as compared to the control group. In contrast, the serum concentration of nitrate/nitrite was significantly higher in the PFLX group 4 h after administration, but there was no difference from controls was observed at 8 and 24 h. These results suggest that NO is involved in the induction of Achilles tendon lesions in juvenile rats by pefloxacin (PFLX) and may be similar to the tendon disorder of humans receiving quinolones. PMID- 9403276 TI - Who cares about pharmacovigilance? PMID- 9403277 TI - University hospital-based drug information service in a developing country. AB - In 1994, a clinically oriented drug information unit was established at the Tribhuvan University Teaching hospital in Nepal, with a view to providing objective and independent information through a question-answer service and bulletin production. During the first 2 years of its service, the unit received a total of 674 encounters, with an average of 28 inquiries a month (range 13-42): about three-quarters (74.5%) of all the inquiries were from prescribing doctors, including 38.0% from specialist clinicians: about a quarter (24.6%) were related to patient problems. Most (86.8%) of the responses were provided within 24 h of the inquiry. Frequently encountered queries related to: pharmacotherapy of a disease or drug indication(s), adverse drug reactions, drug doses, availability, drug use in pregnancy, ingredient(s) of a proprietary product, precautions for use and drug interactions. Details of the inquiries received and the responses provided by the unit are documented in a standard question-answer form. The unit also carries out proactive dissemination of information through the publication and free distribution of a bimonthly bulletin which includes brief referenced reviews on drug- and therapeutics-related topics. Nepal- or the local situation related write-ups are now being increasingly included in the bulletin. A users' survey carried out at the end of 1-year service indicated that the question answer and bulletin production activities of the unit were well-perceived by its target audiences, i.e. the prescribing doctors and postgraduate medical students. Although Medline on CD-ROM and original journal articles available in the hospital library were consulted for answering a few of the questions, the vast majority of them could be adequately handled by consulting a limited number of well-known drug information books. Our experience indicates that in developing countries such as Nepal, where funds are often severely limited, a small-scale drug information centre, serving a local area, can be usefully initiated by a few motivated staff with a modest collection of about a dozen key reference books. PMID- 9403278 TI - Twenty-four-hour blood pressure monitoring during treatment with extended-release felodipine versus slow-release nifedipine in elderly patients with mild to moderate hypertension: a randomized, double-blind, cross-over study. AB - OBJECTIVE: This double-blind, placebo-controlled randomized study was designed to compare the antihypertensive effect and tolerability of extended-release felodipine and slow-release nifedipine retard in elderly hypertensive patients. METHODS: Thirty patients of both sexes (mean age 71 years) with mild to moderate essential hypertension were recruited from our hypertension outpatient clinic. After a 2-week placebo period, felodipine extended-release (felodipine ER), 10 mg once daily, nifedipine slow-release retard (nifedipine SR), 20 mg twice daily or placebo were administered to each patient for 2 weeks according to a 3 x 3 latin square design. At the end of each treatment period, the patients underwent 24-h noninvasive blood pressure monitoring. RESULTS: All of the patients completed the trial and no serious adverse experience was reported. In comparison with placebo, felodipine and nifedipine decreased mean 24-h diastolic blood pressure by 6.7 and 4.3 mmHg, respectively, with no significant difference between the two drugs. Mean 24-h systolic blood pressure also decreased after felodipine and nifedipine, with no difference between the two drugs. Both drugs reduced blood pressure variability, lowering the 24-h mean standard deviation of mean hourly blood pressure values. The trough:peak ratio for felodipine was 80% for systolic and 75% for diastolic blood pressure. CONCLUSION: Felodipine ER once daily lowers blood pressure in elderly hypertensives and is as effective as nifedipine SR twice daily. The high trough:peak ratio suggests that the dose and the between dose interval of felodipine provides adequate therapeutic coverage. PMID- 9403279 TI - Povidone-iodine wash solutions in the prevention of superficial fungal infections; predictive evaluation using the corneofungimetry bioassay. AB - OBJECTIVE: Prevention of superficial mycoses remains a stubborn problem. The effect of antiseptics for that purpose is largely unknown. We studied the potential fungitoxic activity of povidone iodine (PVP-I) contained in wash solutions. METHODS: The corneofungimetry bioassay was conducted using PVP-I at 1.33%, 2.5%, 4% and 7.5% as test products and four target fungi, namely Candida albicans, Trichophyton rubrum, T. mentagrophytes, var. interdigitale (20 strains) and Microsporum canis. RESULTS: Data show that PVP-I limits fungal growth on stratum corneum. Different species and strains of fungi are not similarly affected. There also exists a diversity of individual susceptibility of the stratum corneum to promote germination of yeasts and arthroconidia. PMID- 9403280 TI - Drug-induced chest pain and myocardial infarction. Reports to a national centre and review of the literature. AB - OBJECTIVES: To analyse reports of drug-induced myocardial infarction and chest pain sent to a national reporting centre. To review which drugs were suspected of exhibiting these adverse events and what mechanisms were involved. METHODS: During the 20-year period 1975 through 1994, a total of 19,141 reports on adverse reactions to drugs were received by the Netherlands Centre for Monitoring of Adverse Reactions to Drugs. Of these 19,141 reports, 220 (1.1%) were concerned with drug-induced chest pain or myocardial infarction. After excluding reports in which the causal relationship was unlikely, poorly documented reports and reports on cases of overdosage, 183 reports (84%) were analysed. RESULTS: There were 130 reports (71%) of drug-induced chest pain and 53 reports (29%) of drug-induced myocardial infarction. A total of 104 reports concerned females (57%). The most frequently reported suspected drugs were the antimigraine drug sumatriptan (33 reports, 4 concerning myocardial infarction), the calcium antagonist nifedipin (9 reports, 2 of myocardial infarction) and nicotine [9 reports (8 patches, 1 chewing gum), 5 concerning myocardial infarction]. There were 18 reports of a fatal outcome. CONCLUSIONS: Several drugs can produce chest pain or myocardial ischaemia. It is important to recognise drugs as a potential cause, especially in patients with normal coronary arteries. PMID- 9403281 TI - Pharmacokinetics of mezlocillin and sulbactam under continuous veno-venous hemodialysis (CVVHD) in intensive care patients with acute renal failure. AB - OBJECTIVE: In intensive care medicine, continuous detoxication methods, such as continuous veno-venous hemodialysis (CVVHD), are used for treating acute renal failure. However, in contrast to conventional hemodialysis, little is known about the pharmacokinetics of many drugs administered in this setting and guidelines for dosages of drugs often do not exist. This holds particularly true for broad spectrum antibiotics, which are often required during intensive care. METHODS: In this study, we investigated the pharmacokinetics of the acylureidopenicillin mezlocillin and the beta-lactamase inhibitor sulbactam during CVVHD and deduced dosage recommendations from the kinetic parameters with the goal of maintaining trough levels of above 10 mg.l-1 for mezlocillin and 5 mg.l-1 for sulbactam. Six intensive care patients with acute renal failure, receiving mezlocillin (n = 5) and/or sulbactam (n = 4), were examined during CVVHD and during intervals between CVVHD. The serum concentrations and the amounts of the drugs excreted into the dialyzate and into the urine within one dosage interval were measured using high performance liquid chromatography (HPLC). Three of the patients were jaundiced, indicating functional impairment of the liver. RESULTS: The clearances by CVVHD (CLCVVHD) for mezlocillin ranged between 11.0 and 44.9 ml.min-1 and the half lives ranged between 1.12 and 8.84 h. Low CL and long half lives were observed in the patients with jaundice. For sulbactam, CLCVVHD ranged between 10.1 and 22.8 ml.min-1 and serum half lives were 4.25-6.11 h, independent of liver function. CONCLUSION: Due to high hepatobiliary clearance of mezlocillin, dosage adjustments in patients with acute renal failure, treated by CVVHD, are needed only with concurrent impaired liver function. For sulbactam, the optimal dose was found to be 0.5 g, administered every 12 h, regardless of liver function. PMID- 9403282 TI - The pharmacokinetics of pravastatin in patients on chronic hemodialysis. AB - OBJECTIVE: The single-dose and steady-state pharmacokinetics of the HMG CoA reductase inhibitor pravastatin and its two metabolites, SQ 31,906 and SQ 31,945, were evaluated in 12 hemodialysis patients. A single 20-mg i.v. dose was employed, followed by daily oral dosing of 20 mg over four hemodialysis intervals. RESULTS: No statistical differences in the pharmacokinetics of pravastatin or SQ 31,906 were evident when comparing the first and last days of oral dosing with pravastatin. The pharmacokinetic parameters of pravastatin and SQ 31,906 were similar to those of healthy volunteers. SQ 31,945, the inactive polar metabolite, did accumulate in dialysis patients, as evidenced by an accumulation index of 1.7 +/- 1.0. Although metabolic clearance is the predominant mode of elimination of pravastatin, hemodialysis clearances of pravastatin, SQ 31,906 and SQ 31,945 will contribute to total body clearance since dialytic clearance ranged from 40 to 80 ml.min-1. CONCLUSION: Pravastatin can be safely administered in the usual dosages to subjects with renal failure on hemodialysis and no change in dosing is necessary. PMID- 9403283 TI - Clearance of ceftriaxone during haemodialysis using cuprophane, haemophane and polysulfone dialysers. AB - OBJECTIVE: The purpose of the present study was to investigate whether the clearance of ceftriaxone during haemodialysis is influenced by the type of membrane used (cuprophane, haemophane or polysulphone). METHODS: After administration of a single 2-g dose of ceftriaxone, the half-life of the drug during haemodialysis and the clearance of the dialyser were measured. RESULTS: The mean dialysis clearance normalised for square metre of membrane surface was significantly different for the three dialysers (haemophane 24 ml.min-1.m-2; cuprophane 32 ml.min-1.m-2; polysulphone 42 ml.min-1.m-2). CONCLUSIONS: Polysulphone membranes are more permeable and increase the extraction of ceftriaxone more than the other dialysers studied (haemophane and cuprophane membranes). These results, taken together with previous data, show that an increase of the dose in dialysis patients treated with large surface (> 0.8 m2) and high permeability membranes might be necessary. PMID- 9403284 TI - Comparative pharmacokinetics of dirithromycin and erythromycin in normal volunteers with special regard to accumulation in polymorphonuclear leukocytes and in saliva. AB - OBJECTIVE: In a randomized cross-over study, we assessed pharmacokinetics and intracellular concentrations in polymorphonuclear leukocytes (PMN) and saliva of erythromycin and erythromycylamine, the active metabolite of dirithromycin. METHODS: Ten healthy volunteers received 1 g erythromycin b.i.d. or 500 mg dirithromycin qd for 5 days (wash out period, 35 days). Concentrations of erythromycin and erythromycylamine were measured in serum, urine, saliva, and granulocytes by bioassay and high-performance liquid chromatography (HPLC) on days 1, 3, and 5 of each study period, respectively. RESULTS: While maximal serum concentrations (Cmax) and the area under the data (AUDtot) of erythromycin were significantly higher (Cmax 1.44 mg.l-1, AUDtot 5.66 mg.h.l-1) than those of erythromycylamine (Cmax 0.29 mg.l-1, AUDtot 1.96 mg.h.l-1), erythromycylamine had a significantly higher mean residence time (21 h) than erythromycin (5.5 h). Erythromycylamine accumulated significantly more in PMN than erythromycin; the accumulation factor of erythromycylamine was 100 with a maximal intracellular concentration of 13.4 mg.l-1, whereas the maximal accumulation factor of erythromycin was 4 with a maximal intracellular concentration of 6.1 mg.l-1. There were no significant differences in maximal saliva concentrations (erythromycin 0.35 mg.l-1, erythromycylamine 0.31 mg.l-1). PMID- 9403285 TI - Food increases the bioavailability of mefloquine. AB - OBJECTIVES: To assess the effect of food on the pharmacokinetics of the antimalarial mefloquine and its major plasma metabolite in healthy volunteers. METHODS: In an open, two-way cross-over study, 20 healthy male volunteers who had fasted overnight were randomised to receive a single oral dose of 750 mg mefloquine in the absence or presence of a standardised, high-fat breakfast, administered 30 min before drug administration. Blood samples were taken at specific times over an 8-week period. Plasma concentrations of mefloquine and its carboxylic acid metabolite were determined by high-performance liquid chromatography for pharmacokinetic evaluation. RESULTS: The parameters Cmax and AUC of both mefloquine and its metabolite were significantly (P < 0.05) higher under post-prandial conditions than under fasting conditions (mefloquine: mean Cmax 1500 vs 868 micrograms.l-1, mean AUC 645 vs 461 mg l-1.h; metabolite: Cmax 1662 vs 1231 micrograms.l-1, AUC 1740 vs 1310 mg l-1.h). The intersubject variability in Cmax and AUC of mefloquine was less than 30% (coefficient of variation). The time to peak plasma concentration of mefloquine was significantly shorter after food intake (17 vs 36 h). Compared with absorption in volunteers who had fasted, food did not alter t1/2 (mefloquine and its metabolite) and tmax (metabolite). CONCLUSION: Under the conditions of this study, food increases the rate and the extent of mefloquine absorption. It is reasonable to recommend that mefloquine be administered with food in travellers receiving chemoprophylaxis and in patients on recovery receiving curative treatment. In acutely ill patients, mefloquine should be taken as soon as possible and administration with or shortly after meals should be attempted as soon as feasible. PMID- 9403286 TI - The influence of two types of meal on the pharmacokinetics of a modified-release formulation of nifedipine (Adalat Retard). AB - OBJECTIVE: The effect of two different types of meal on the absorption of a modified-release formulation of nifedipine (Adalat Retard) was studied. RESULTS: A light breakfast produced a delay in gastric emptying (indicated by the rate of paracetamol absorption) compared with the fasting state but did not alter the tmax or Cmax for nifedipine significantly. After a cooked breakfast, there was less delay in gastric emptying and again no delay in tmax for nifedipine. However, the Cmax for nifedipine was significantly higher than in the fasting state. Neither meal influenced the bioavailability of nifedipine. CONCLUSION: The results suggest that the nature of the meal has an important influence on the absorption profile of this formulation of nifedipine, probably by an effect on its dissolution. This study illustrates the importance of considering the effects of different types of meal before concluding that food does not affect the pattern of drug absorption. PMID- 9403287 TI - The effect of codeine on gastrointestinal transit in extensive and poor metabolisers of debrisoquine. AB - METHODS: Codeine (50 mg) was administered to 12 extensive metabolisers (EM) and 12 poor metabolisers (PM) of debrisoquine. The oro-caecal transit time was estimated by the hydrogen breath test. The urinary excretion of codeine and metabolites during a 6-h interval was estimated after simultaneous analysis of codeine, morphine-3-glucuronide (M3G), morphine-6-glucuronide (M6G), morphine (M), normorphine (NM), norcodeine, norcodeine glucuronide and codeine-6 glucuronide using HPLC. RESULTS: The mean transit times after placebo were 1.3 h in the EM and 1.4 h in the PM. The corresponding figures after ingestion of codeine were 2.2 h and 2.1 h. The differences between the groups were statistically and clinically insignificant. The effect of codeine compared with placebo was significantly different in both groups. As expected, the metabolites of the O-demethylation pathway, M, M6G, M3G and NM were significantly lower in the PM. Interestingly, the recovery of the dose in the form of codeine (> 1.7 times) and norcodeine (> 2.5 times) was significantly higher in the PM, indicating compensatory metabolism via N-demethylation. CONCLUSION: In contrast to the analgesic effect, the prolongation of gastrointestinal transit caused by the drug does not depend on the formation of O-demethylated active metabolites M, M6G or NM. PMID- 9403288 TI - Concomitant administration of the alpha-glucosidase inhibitor voglibose (AO-128) does not alter the pharmacokinetics of glibenclamide. AB - OBJECTIVE: Voglibose is a new and potent inhibitor of alpha-glucosidases used for treatment of diabetes mellitus. It increases gastro-intestinal motility and could thus affect absorption of other concurrently administered antidiabetic drugs. The aim of this study was to investigate whether or not voglibose modifies the pharmacokinetics of glibenclamide, a widely used oral antidiabetic, and the glibenclamide-induced decrease in fasting serum glucose. METHODS: Twelve healthy male subjects were included in this double-blind cross-over study and received a single 1.75-mg dose of glibenclamide on the 8th day of continuous administration of either placebo (reference) or voglibose 5 mg t.i.d. (test). Blood samples were taken to determine the pharmacokinetic characteristics of glibenclamide and the test/reference ratios were evaluated according to bioequivalence criteria. Additional blood samples were taken to measure serum glucose on the same day. RESULTS: The concentration-time course of glibenclamide under concomitant voglibose administration was similar to that under placebo. The equivalence ratio (test/reference) for the pharmacokinetic characteristics AUCnorm was 1.03 (geometric mean; 0.95-1.11, 90% confidence interval) and Cmax.norm 1.01 (0.94 1.08). The parameters were within the accepted range of 0.8-1.25 (AUC) or 0.7 1.43 (Cmax), thus fulfilling equivalence criteria and indicating no effect of voglibose on glibenclamide kinetics. The glibenclamide-induced decrease in fasting serum glucose concentration was similarly independent of placebo or voglibose co-administration. CONCLUSIONS: Voglibose did not interact with glibenclamide on a pharmacokinetic level. Concomitant treatment was well tolerated and has been proven to be safe for further clinical use. PMID- 9403289 TI - The alpha-glucosidase inhibitor voglibose (AO-128) does not change pharmacodynamics or pharmacokinetics of warfarin. AB - OBJECTIVE: Voglibose is a new and potent inhibitor of alpha-glucosidases and is used for the treatment of diabetes mellitus. Since voglibose increases gastrointestinal motility and could thus affect absorption of concomitantly administered drugs, it was investigated whether or not voglibose modifies the pharmacodynamics and pharmacokinetics of warfarin, an oral anticoagulant frequently used in cardiovascular disorders likely to arise in diabetic patients. METHODS: Twelve healthy male subjects were given individually adjusted doses of warfarin to reduce prothrombin time (Quick's method) to a value of about 30-40% of the normal range within the first 8 days. Then, the individuals maintenance dose, given in the morning, was maintained until day 15. On study days 11-15, voglibose was co-administered per os in a dose of 5 mg t.i.d. The prothrombin time was determined on days 10 and 11 (reference) and on days 15 and 16 (test), and the steady-state pharmacokinetic characteristics of the warfarin enantiomers were determined on days 10 (reference) and 15 (test). The ratios test/reference were evaluated according to bioequivalence criteria. RESULTS: The equivalence ratio (test reference) for the pharmacodynamic parameter prothrombin time was 0.97 and for the pharmacokinetic characteristics AUC0-24 h.t.ss: S-(-)-warfarin, 1.05; R-(+)-warfarin, 1.01; and Cmax.ss: S-(-)-warfarin, 1.08; R-(+)-warfarin, 1.04. All parameters were within the predetermined accepted range of 0.7-1.43 (pharmacodynamics) or 0.8-1.25 (pharmacokinetics), thus fulfilling equivalence criteria. CONCLUSIONS: Voglibose modified neither the pharmacodynamics nor the pharmacokinetics of warfarin under steady-state conditions. Concomitant treatment was well tolerated and has been proven to be safe for further clinical use. PMID- 9403290 TI - Tramadol withdrawal in a neonate. PMID- 9403291 TI - Internal fixation sets for use in the hand. A comparison of available instrumentation. AB - Optimal clinical results of internal fixation in the hand depend on appropriate patient selection criteria, good surgical techniques, and appropriate implant selection. A variety of implant sets with plates and screws from different manufacturers are available for hand bone fixation. Traditionally, such sets have been made of stainless steel, but more recently, implants have been manufactured from titanium. Sizes and configurations vary significantly between manufacturers. In this article, commonly available internal fixation sets are compared. Plate, screw, and instrument design are considered. These comparisons should help the surgeon make an enlightened choice of implant sets. PMID- 9403292 TI - Fixation of phalangeal fractures. AB - The indications and methods for operative treatment of phalangeal fractures are reviewed. Included are descriptions with examples of closed reduction and percutaneous pin fixation, percutaneous reduction and percutaneous fixation, open reduction and internal fixation, and static and dynamic external fixation. Also included are tips for facilitating phalangeal fixation and a description of potential complications. PMID- 9403293 TI - Metacarpal fixation. AB - This article discusses and categorizes common metacarpal fractures and their treatments, including various techniques of obtaining skeletal fixation. It reviews metacarpal shaft fracture; intra-articular metacarpal head fractures; and metacarpal neck and base fractures including carpal-metacarpal dislocations, and Rolando's and Bennett's fractures of the base of thumb metacarpal. Also discussed are the effects of shortening of lengthening the digital skeleton and bioabsorbable implants, a potential treatment modality currently under development. PMID- 9403294 TI - Internal fixation of scaphoid fractures. AB - Scaphoid fracture fixation is indicated in certain acute situations and for scaphoid nonunion. Internal fixation requires an understanding of the distinctive scaphoid anatomy with relation to its shape, blood supply, fracture healing, and radiographic evaluation. The choice of surgical approach varies with the fracture configuration and procedure planned. A variety of innovative implants are available to accomplish internal fixation of the scaphoid. The ultimate goal is to achieve union and restore stability to the carpus. PMID- 9403295 TI - Fractures of the carpal bones. AB - Fractures of the carpal bones, excluding the scaphoid, are less common and are often missed on standard, plain radiographs. The diagnosis requires knowledge of the anatomy and common fracture patterns of the bones and the specialized radiographic views necessary to image them. Although the hamate hook, trapezial ridge, and pisiform often fracture from direct trauma, other fractures of the carpus may indicate more widespread injury and require a detailed evaluation to rule out associated perilunate or carpometacarpal fracture-subluxations. Fortunately, if treated appropriately, the vast majority of these fractures heal uneventfully and allow recovery of motion and function of the wrist and hand. PMID- 9403296 TI - Internal fixation for small joint arthrodeses in the hand. The interphalangeal joints. AB - Interphalangeal arthrodeses are performed frequently for disabling pain and deformity in the proximal and distal interphalangeal joints. Such arthrodeses are the gold standard for eliminating pain in arthritic joints of the fingers. Kirschner wire fixation is simple to perform but often leads to nonunion or delayed union. Fixation with tension band wires and Herbert and Accutrak screws is discussed in this article. The techniques are simple and straightforward, and they afford considerably more stability and a higher union rate than Kirschner wires alone. PMID- 9403298 TI - Static external fixation in the hand and carpus. AB - Static external fixation in the hand and carpus remains a valuable part of the hand surgeon's armamentarium for the treatment of complex injuries. This article describes a brief history, indications, case examples, and complications using these constructs. Pertinent pitfalls are emphasized to avoid suboptimal outcomes. PMID- 9403297 TI - Limited wrist fusion. AB - Limited wrist fusion can provide patients a measure of pain relief with retention of a functional range of motion. Recent biomechanic investigations have shed light on the potential benefits of such procedures. Clinical studies have demonstrated the usefulness of limited wrist fusion for the treatment of numerous conditions involving the wrist. Significant complications are associated with many of these fusions and should be considered before performing these procedures. PMID- 9403299 TI - Dynamic skeletal fixation in the upper extremity. AB - Recent advances in the understanding of the biomechanics of the elbow and improvement in material designs have revolutionized the ability to treat difficult problems of the proximal interphalangeal and elbow joints. Dynamic skeletal fixation addresses the desire to allow an injured joint to heal, move, and remain stable simultaneously. In the past, such goals were achieved sequentially or in series. It is now possible to perform such tasks in parallel. PMID- 9403300 TI - Fixation for distal radius fractures. AB - Distal radius fractures are common fractures that can cause significant disability. As techniques and implants have improved, better results can be expected from internal and external fixation of complex wrist fractures. With meticulous technique, articular alignment can be secured and small articular fragments can be replaced in anatomic locations through limited open or arthroscopic techniques. Bone graft should be used liberally in comminuted articular fractures. Such procedures are demanding, however. Familiarity with the techniques described in this article will enhance the surgeon's ability to restore function in this group of patients. PMID- 9403301 TI - Wrist fusion. AB - Wrist arthrodesis is a reliable procedure for the treatment of a variety of disorders of the wrist. It provides predictable pain relief, enhanced hand function, and a high degree of patient satisfaction. The AO/ASIF wrist fusion plate allows rigid internal fixation and optimizes wrist position for maximum hand function. In comparison to other wrist arthrodesis techniques, the wrist fusion plate produces a high rate of fusion utilizing local bone graft from the distal radius. PMID- 9403302 TI - Forearm fixation. AB - The great majority of forearm fractures in adults are best treated by open reduction and internal fixation. Although alternative methods exist, plate fixation is favored by most surgeons. With strict attention to surgical detail, complication rates are low and early active function is possible. The treatment of high-energy, open fractures can include various techniques such as internal or external fixation. Refracture remains the greatest risk following hardware removal, which is not necessary for all patients. PMID- 9403303 TI - Fixation of complex elbow fractures, Part I. General overview and distal humerus fractures. AB - Complex elbow fractures are those intra-articular injuries which result in displacement or incongruity of one or more of the articulations between the distal humerus, proximal radius, or proximal ulna. The article discusses a system for clinical and radiographic evaluation, operative management, and rehabilitation of distal humerus fractures. This review presents detailed descriptions of skeletal reconstructive techniques for these injuries based on a review of the literature, biomechanic principles, and clinical experience. The importance of rigid internal fixation and early postoperative mobilization is emphasized. PMID- 9403304 TI - Fixation of complex elbow fractures, Part II. Proximal ulna and radius fractures. AB - Complex fractures of the elbow often include significant injury to the proximal ulna and/or proximal radius. Such injuries are often combined with injury to the periarticular soft tissues. Appropriate treatment is dependent on accurate diagnosis, definitive treatment of both the skeletal and ligamentous components of injury, and initiation of rehabilitation programs the stress early motion. PMID- 9403305 TI - Fixation methods in contaminated wounds and massive crush injuries of the forearm. AB - This article presents a critical review of the current literature on the stabilization methods of severe open fractures or crush injuries of the forearm. Due to the complexity of the problem, the advantages and disadvantages of the different fixation methods are presented in order to provide the reader with a combination of possibilities for timing and decision-making when confronted with such fractures in clinical practice. PMID- 9403306 TI - Infection in the presence of skeletal fixation in the upper extremity. AB - Infection is infrequent after open fractures of the upper extremity. Treatment begins with prevention through the appropriate use of prophylactic antibiotics, adequate wound debridement, and timely soft-tissue coverage. If infection supervenes, the surgeon must identify the responsible bacteria and administer antibiotics accordingly. Stable fixation is retained. Unstable fixation is removed and skeletal stability restored. Using these principles, infection in the presence of skeletal fixation can be controlled and fracture union achieved. PMID- 9403307 TI - Therapy after skeletal fixation in the hand and wrist. AB - Skeletal fixation can be very beneficial to the recovery of wrist and hand function after a displaced fracture because it allows mobilization of soft tissues before the completion of fracture healing. The benefits of skeletal fixation can be greatly diminished, however, if excessive force causes the fixation to fail before fracture healing has occurred, infection occurs around the implant, or the patient develops reflex sympathetic dystrophy. Those complications, as well as others, are often caused by inappropriate or inadequate hand therapy. This article discusses the techniques needed to avoid many such complications while providing the best possible functional result for every patient. PMID- 9403308 TI - Wave-intensity analysis: a new approach to left ventricular filling dynamics. AB - In order to explore a new approach to the analysis of diastolic dysfunction, we adapted wave-intensity analysis (WIA), a time-domain analysis that provides information regarding both upstream and downstream events, to left ventricular (LV) filling. WIA considers the pressure and flow waves as summations of successive wavelets, characterised by the direction they travel and by the sign of the pressure gradient associated with them. Wave intensity is the product, dPdU, calculated from the incremental differences in LV pressure (dP) and mitral velocity (dU) and, during the diastolic filling interval, yields up to five dPdU peaks. Peak 1 is caused by backward-travelling expansion waves that accelerate the blood while LV pressure falls, and may be related to "diastolic suction". Peak 2 is caused by forward-travelling compression waves which occur if acceleration continues after LV pressure begins to increase. Peak 3 is caused by backward compression waves and is associated with rising LV pressure and deceleration. Peak 4 is caused by forward compression waves and is associated with the increasing LV pressure and acceleration caused by atrial contraction. Peak 5 is caused by backward compression waves and is associated with increasing pressure and deceleration. These preliminary observations suggest that WIA can be useful in describing the mechanics of LV filling and, after much further work has been accomplished, it might prove useful in the detection and characterization of diastolic dysfunction. PMID- 9403309 TI - Anti-atherogenicity in women does not prevent restenosis after balloon angioplasty. AB - To test the hypothesis that anti-atherogenicity in women exerts beneficial effects to prevent restenosis formation after coronary angioplasty, we studied 493 men (988 lesions) and 81 women (159 lesions), aged 40-60 years, who had undergone successful balloon angioplasty and had follow-up angiography, 4.9 +/- 4.1 months later. We compared the extent of restenosis between men and women, and between pre- and post-menopausal women, which was assessed by a categorical definition of restenosis (more than 50% diameter stenosis at follow-up) and by percent diameter measured immediately after angioplasty and at follow-up. Hypertension was more frequent in women and a significantly lower percentage of women smoked. In women, the levels of total cholesterol and low-density lipoprotein cholesterol were higher. The location of dilated lesions, frequency of angioplasty for lesions with chronic total occlusion, and frequency of emergency angioplasty in patients with unstable angina or acute myocardial infarction were similar in men and women. Restenosis formation, estimated by the categorical definition or percent diameter, did not differ between men and women, or between pre- and post-menopausal women. Menopausal status or sex was not an independent predictor of restenosis by multivariate analysis. Thus, the benefit of anti-atherogenicity in women does not play an important role in preventing restenosis after coronary angioplasty. PMID- 9403310 TI - Effects of levamisole, an immunomodulator, upon murine encephalomyocarditis virus myocarditis. AB - To test the therapeutic efficacy of levamisole, 5-week-old DBA/2 mice were inoculated intraperitoneally with 10 plaque-forming units of encephalo myocarditis virus. Levamisole (2.5 mg/kg/per day) was administered intraperitoneally daily, starting simultaneously with the virus inoculation, in experiment I for 14 days, and daily on days 14 to 28 in experiment II in mice that survived to 14 days after virus inoculation. In experiment I, survival was higher, the severity of myocarditis was less, and myocardial virus titers were lower in treated than in untreated animals. In experiment II, levamisole was not effective. No significant changes in serum neutralizing antibody titers occurred in either experiment. Furthermore, levamisole prevented associated lymphoid organ atrophy induced by the virus infection. An additional in vitro study revealed the absence of anti-viral activity of the drug. Thus, levamisole may have favorable effects upon encephalomyocarditis virus myocarditis by preventing the virus induced lymphoid organ atrophy and reducing myocardial virus replication in the acute stage. PMID- 9403311 TI - Activation of cardiac muscarinic receptor and ischemic preconditioning effects in in situ rat heart. AB - Activation of cardiac muscarinic receptors by vagal stimulation decreases cardiac work, which may have a protective effect against ischemic injury. To determine whether cardiac muscarinic receptors contribute to the mechanisms of preconditioning effects, we examined the effect of carbachol on ischemia/reperfusion damage and the effect of vagotomy on cardioprotection induced by ischemic preconditioning. Rats were subjected to 30 min of left coronary artery occlusion followed by 30-min reperfusion in situ. Pre conditioning was induced by three cycles of 2-min coronary artery occlusion and, subsequently by 5 min of reperfusion. The incidence of ischemic arrhythmias, such as ventricular tachycardia (VT) and ventricular fibrillation (VF), and the development of myocardial infarction were markedly reduced by the preconditioning. Carbachol infusion (4 micrograms/kg per min) delayed the occurrence of VT and VF during ischemia and reduced the infarct size. Compared with non-ischemic left ventricle, the cyclic guanosine monophosphate (GMP) content in the ischemic region of the left ventricle was decreased by ischemia/reperfusion, whereas the cyclic adenosine monophosphate (AMP) content of this region was increased. These changes were reversed by preconditioning. Similar changes in cyclic GMP and AMP content in the ischemic region were seen in rats undergoing carbachol treatment. These results suggest the possible contribution of muscarinic receptor stimulation to preconditioning. Vagotomy prior to preconditioning diminished the antiarrhythmic effects, whereas it did not block the anti-infarct effect afforded by pre-conditioning. Vagotomy abolished the preconditioning effect on the tissue cyclic GMP, but it did not attenuate the decrease in tissue cyclic AMP. The results suggest that muscarinic stimulation exerts preconditioning-mimetic protective effects in ischemic/reperfused hearts, but that a contribution of reflective vagal activity to the mechanism for preconditioning is unlikely. PMID- 9403313 TI - Flow characteristics and required control algorithm of an implantable centrifugal left ventricular assist device. AB - As the clinical application of LVADs has increased, attempts have been made to develop smaller, less expensive, more durable and efficient implantable devices using rotary blood pumps. Since chronic circulatory support with implantable continuous-flow LVADs will be established in the near future, we need to determine the flow characteristics through an implantable continuous-flow LVAD. This study describes the flow characteristics through an implantable centrifugal blood pump as a left ventricular assist device (LVAD) to obtain a simple non invasive algorithm to control its assist flow rate adequately. A prototype of the completely seal-less and pivot bearing-supported centrifugal blood pump was implanted into two calves, bypassing from the left ventricle to the descending aorta. Device motor speed, voltage, current, flow rate, and aortic blood pressure were monitored continuously. The flow patterns revealed forward flow in ventricular systole and backward flow in diastole. As the pump speed increased, an end-diastolic notch became evident in the flow profile. Although the flow rate (Q [l/min]) and rotational speed (R [rpm]) had a linear correlation (Q = 0.0042R 5.159; r = 0.96), this linearity was altered after the end-diastolic notch was evident. The end-diastolic notch is considered to be a sign of the sucking phenomenon of the centrifugal pump. Also, although the consumed current (I [A]) and flow rate had a linear correlation (I = 0.212Q + 0.29; r = 0.97), this linearity also changed after the end-diastolic notch was evident. Based upon the above findings, we propose a simple algorithm to maintain submaximal flow without inducing sucking. To maintain the submaximal flow rate without measuring flow rate, the sucking point is determined by monitoring consumed current according to gradual increases in voltage. PMID- 9403312 TI - Effects of methylprednisolone on hemodynamics and beta-adrenergic receptor signaling in rabbits with acute left ventricular failure. AB - Studies suggest that corticosteroids may restore the responsiveness to catecholamines in hypotensive patients. Since the significance of this promising intervention in congestive heart failure remains to be explored, we determined the effects of methylprednisolone, a potent activator of beta-adrenergic receptor signaling, on hemodynamics and beta-adrenergic receptor regulation in an animal model of heart failure. Acute left ventricular overloading was produced by aortic regurgitation (AR) in 22 Japanese white rabbits. Eleven animals received an intravenous administration of methylprednisolone (AR + PSL), while 11 received saline (AR + C) for 1 week. A sham operation was performed on 10 other rabbits (S). There was no difference between the AR + C and AR + PSL groups in the decrease in aortic diastolic pressure immediately after the production of AR. The aortic diastolic pressure and regurgitant fractions were also similar in the two groups. The left ventricular end-diastolic and end-systolic dimensions were both larger, and the left ventricular end-diastolic pressure was higher in AR + C or AR + PSL than in S rabbits. Between the AR + C and AR + PSL, there were no differences in any of these variables. Cardiac output was lower in AR + C, but not in AR + PSL, than in S. Cardiac output in AR + PSL was significantly higher than in AR + C. The myocardial concentration of norepinephrine and the number of beta-adrenergic receptors were both lower in the AR + C and AR + PSL than in the S groups. The number of receptors in AR + PSL was higher than in AR + C. Maximal isoproterenol-stimulated adenylyl cyclase activity was similar in the AR + C and AR + PSL groups. Results suggest methylprednisolone yielded some benefits, including an increase in cardiac output and in total beta-adrenergic receptor number, in this animal model of heart failure. PMID- 9403314 TI - Myocardial histological changes in dilated cardiomyopathy during a long-term left ventricular assist device support. AB - As the myocardium in patients with dilated cardiomyopathy (DCM) is deteriorating progressively, resulting in a decrease in left ventricular function, patients with end-stage DCM may require implantation of a left ventricular assist device (LVAD) unless they undergo heart transplantation. Although LVAD has been reported to provide excellent hemodynamic support, no data are currently available about the effects of long-term LVAD support on the myocardium in patients with DCM. We describe two patients with end-stage DCM who underwent LVAD implantation and were supported with LVAD for 524 and 245 days, respectively. Serial myocardial biopsies showed increases in myocardial cell diameter and intercellular fibrosis, despite excellent hemodynamic support by LVAD. These data suggest that the myocardium in patients with end-stage DCM deteriorates progressively, even if the pre-load of the left ventricle is reduced by LVAD. PMID- 9403315 TI - Weight, nutrition and hormonal events in women. Introduction. PMID- 9403316 TI - Genetic determinants of regional fat distribution. AB - Upper body fat and abdominal visceral fat are two obesity-related phenotypes of interest because of their relationships with a variety of metabolic complications. The heritability of the amount of upper body fat or the level of upper body fat relative to lower body fat ranges from approximately 30-50% of the phenotype's age, sex and total body fat adjusted variance. On the other hand, familial studies of abdominal visceral fat reveal that the familial transmission reaches > 50% of the age, sex and total body fat adjusted variance. Complex segregation analysis undertaken with a panel of nuclear families indicates that major genes may account for a significant fraction of the variance in upper body fat and abdominal visceral fat. Two intervention studies conducted with pairs of male identical twins have shown that changes in upper body fat and visceral fat are more similar within pairs than between pairs, either in phenotype increments when challenged by chronic overfeeding, or in adipose tissue losses after exposure to long-term negative energy balance conditions. The evidence accumulated to date is sufficient to justify undertaking a search for the specific genes and molecular markers involved in the heterogeneity commonly observed in human fat topography. PMID- 9403317 TI - Adrenergic regulation of adipocyte metabolism. AB - Five adrenoceptor (AR) subtypes (beta 1, beta 2, beta 3, alpha 2 and alpha 1), are involved in the control of white and brown fat cell function. A number of metabolic events are controlled by the adrenergic system in fat cells. The stimulatory effect of catecholamines on lipolysis and metabolism is mainly connected to increments in cAMP levels, cAMP protein kinase activation and phosphorylation of various target proteins. Norepinephrine and epinephrine operate through differential recruitment of alpha 2- and beta-AR subtypes on the basis of their relative affinity for the different subtypes (the relative order of affinity is alpha 2 > beta 1 > or = beta 2 > beta 3 for norepinephrine). Antagonistic actions at the level of cAMP production exist between alpha 2- and beta 1-, beta 2- and beta 3-AR-mediated lipolytic effects in human white fat cells. The role of fat cell alpha 2-ARs, which largely outnumber beta-ARs in fat cells of certain fat deposits, in human and primate has never been clearly understood. The other AR type which is not linked to lipolysis regulation, the alpha 1-AR, is involved in the control of glycogenolysis and lactate production. Pharmacological approaches using in-situ microdialysis and selective alpha 2- and beta-AR agonists and antagonists have revealed sex- and tissue-specific differences in the adrenergic control of fat cell function and nutritive blood flow in the tissue surrounding the microdialysis probe. PMID- 9403319 TI - Obesity and reproduction. AB - Obesity produces a variety of alterations in the reproductive system and, similarly, manipulations of the hypothalamic-pituitary-gonadal axis produce changes in food intake, body weight and fat distribution. In men, the primary effects of obesity are a weight related reduction in testosterone and, with massive overweight, a reduction in free testosterone. In females, the weight related development of menarche leads to earlier menarche in obese girls than in normal weight girls. One explanation for the relationship of fatness to menarche may be the ob protein (leptin) which is defective in the obese (ob/ob) mouse. Leptin is secreted by adipose tissue in proportion to the quantity of fat and may serve as a signal to the hypothalamus that fat stores are adequate to nourish a conceptus to term. In women, parity affects obesity and obesity in turn affects the regularity of the menstrual cycle. In many experimental animals with obesity, particularly the genetic forms of obesity, there is complete infertility in the females and marked impairment of reproductive function in the males. In animals with hypothalamic lesions, there is a gender effect on the magnitude of weight gain associated with the sexually dimorphic regions in the medial preoptic area. Castration with removal of oestrogen is followed by obesity in female animals and this can be prevented, as can most forms of obesity, by adrenalectomy. The inhibitory effects of oestrogen on food intake may result from suppression of neuropeptide-Y or galanin peptidergic systems in the arcuate nucleus or medial preoptic area. PMID- 9403318 TI - Hormonal control of regional fat distribution. AB - Hormones exert powerful influences on body fat distribution in humans. Studies under fully controlled conditions in vitro have indicated that cortisol and insulin facilitate lipid accumulation by expressing lipoprotein lipase (LPL). Growth hormone (GH) abolishes this and turns metabolism towards lipid mobilization. Testosterone and GH inhibit LPL and stimulate lipolysis markedly. Cortisol effects are mediated via a glucocorticoid receptor, and testosterone effects via an androgen receptor, the density of which appears to be higher in visceral than subcutaneous adipose tissue. The receptor-mediated effects are probably expressed via transcription of appropriate genes. The female sex steroids also regulate adipose tissue metabolism, but apparently not directly in the absence of specific cellular receptors. Oestrogens seem to exert net effects similar to those of testosterone. These results of cellular studies agree well with in-vivo studies of triglyceride uptake and turnover in different adipose tissue regions. Furthermore, clinical entities with characteristic disturbances in hormone levels show the expected redistribution patterns. PMID- 9403320 TI - Involvement of insulin-like growth factors in the interactions between nutrition and reproduction in female mammals. AB - The effects of nutrition on the reproductive system of female mammals are discussed in an attempt to determine the importance of the role of the insulin like growth factor (IGF) system of proteins in the physiological mechanisms. The IGF system comprises IGF-I, IGF-II, their receptors and binding proteins. For this review, insulin and its receptors have been included in this system. The reproductive responses have been separated into two classes, based on fundamental differences in the reproductive biology and physiological mechanisms underlying them. The first involves the effects of nutrition on the induction of ovulation, at puberty or postpartum. In this case, the question is whether or not the animal will reproduce. The second class of response, changes in ovulation rate, involves consideration of reproductive efficiency in animals in which ovulation is inevitable. PMID- 9403321 TI - Insulin regulation of human ovarian androgens. AB - Hyperinsulinaemic insulin resistance is characteristic of many, if not all, women with polycystic ovary syndrome (PCOS). We will review evidence suggesting that hyperinsulinaemia promotes hyperandrogenism in PCOS by two distinct and independent mechanisms: (i) by increasing circulating ovarian androgens; and (ii) by directly reducing serum sex hormone-binding globulin concentrations. The net result of these actions is to increase circulating free testosterone concentrations. It appears likely that an inherent (probably genetically determined) ovarian defect need be present in women with PCOS, which makes the ovary susceptible to insulin stimulation of androgen production. Limited evidence suggests that hyperinsulinaemia might also promote ovarian androgen production by influencing pituitary release of gonadotrophins. This latter possibility, however, has not been critically evaluated. The clinical implication of these findings is that amelioration of hyperandrogenism in women with PCOS may be achieved by interventions which improve insulin sensitivity and reduce circulating insulin. Such measures might include, but are not limited to, weight loss, dietary modification, and insulin-sensitizing medications. PMID- 9403322 TI - Perspective in the treatment of insulin resistance. AB - Improving the action of insulin is a relatively new concept in therapy. It should, however, become more and more important because of the rapid expansion of the insulin resistance syndrome (including upper body adiposity, glucose intolerance, hypertension, dyslipidaemia, etc.) in industrialized countries and its dramatic consequences for public health. Insulin sensitivity can be improved by non-pharmacological means, essentially reduction of excessive body weight, promotion of regular physical activity and modification of dietary habits, as well as, possibly, cessation of smoking and correction of subclinical magnesium deficiency. Currently available pharmacological means mainly include the biguanide compound metformin and possibly anti-obesity agents, such as (d-) fenfluramine, fluoxetine and benfluorex. New compounds aiming at improving the action of insulin are in development, especially the thiazolidinedione derivatives (e.g. troglitazone), known as 'insulin sensitizers'. Treatment of insulin resistance may have important gynaecological applications, essentially in polycystic ovary syndrome and, possibly, after menopause. Hopefully, improving insulin sensitivity could ameliorate the cardiovascular prognosis of numerous individuals having some or all components of insulin resistance syndrome. PMID- 9403323 TI - Influences of weight, body fat patterning and nutrition on the management of PCOS. AB - Polycystic ovary syndrome (PCOS) is a heterogeneous clinical entity that is defined as the association of hyperandrogenism with chronic anovulation in women without specific underlying diseases of the adrenal or pituitary glands. PCOS is also associated with a metabolic disturbance (insulin resistance). The nature of the complex interrelation of obesity, insulin resistance and endocrine abnormalities in PCOS remains unresolved. However, several studies link obesity, body fat distribution and nutritional habitus with the hormonal and metabolic profiles of PCOS. Moreover, intervention studies have suggested that reducing weight and/or hyperinsulinaemia either by diet alone or by a combination of diet and drugs improves hirsutism, fertility and the hormonal and metabolic profiles of PCOS. In fact, the evaluation of nutritional factors in PCOS is helpful for the screening of metabolic abnormalities and the management of women with PCOS. A point of particular interest in the management of PCOS is that the choice of contraception remains difficult in these high cardiovascular risk women. The impact of pills with ethinyl oestradiol on weight, body fat distribution and carbohydrate metabolism in women with PCOS has not been thoroughly evaluated. The lack of prospective studies to evaluate long-term metabolic and cardiovascular tolerance necessitates care and the assessment of other hormonal possibilities. PMID- 9403324 TI - Weight control and its beneficial effect on fertility in women with obesity and polycystic ovary syndrome. AB - Obesity may be an important pathogenetic factor involved in the development of hyper-androgenism in women with polycystic ovary syndrome (PCOS). Among several other mechanisms, hyperinsulinaemia plays a fundamental role, due to its gonadotrophic function, which has been demonstrated both in vitro and in vivo. Therefore, not surprisingly, weight loss may be expected to have several beneficial effects upon clinical, endocrinological and metabolic features of obese women presenting both PCOS. In particular, weight loss appears to be associated with a significant improvement in menses abnormalities, ovulation and fertility rates, and with a reduction of hyperandrogenism, hyperinsulinaemia, and altered gonadotrophin pulsatile secretion. The central role of improved insulin concentrations and insulin-resistant state is emphasized by the fact that similar effects can be achieved by both short- and long-term administration of metformin, an insulin-lowering drug which ameliorates peripheral insulin action in non diabetic insulin resistant states. We therefore recommend weight loss as a first line therapeutic option in all women with obesity and PCOS. PMID- 9403325 TI - Choice of stimulation in polycystic ovarian syndrome: the influence of obesity. AB - Obesity modifies insulin sensitivity and gonadotrophins dynamics, and is associated with disorders of spontaneous ovulation. High concentrations of leptin are possibly a link between weight and spontaneous ovulation. Weight excess in polycystic ovary syndrome (PCOS) patients increases hyperinsulinaemia, which may result in altered follicular maturation. Obese PCOS women are characterized by a decreased efficiency of the different stimulation treatments. Although clomiphene resistance is not associated with obesity, the dose of clomiphene required to achieve ovulation is positively correlated with body weight. Obese PCOS women also require higher doses of gonadotrophins than their lean counterparts, with ultimately poorer results in pulsatile gonadotrophin releasing hormone-stimulated cycles. The first stage in the optimal management of obese PCOS anovulatory women is a weight loss programme, which helps to correct the clinical and endocrine abnormalities. PMID- 9403326 TI - Deficiency of energy balance and ovulatory disorders. PMID- 9403327 TI - Weight gain in pregnancy. AB - Pregnancy and body weight development are intertwined in complicated patterns. Of the obese patients at our Obesity Unit, 73% had retained > 10 kg in connection with a pregnancy. For the general population the effect of a pregnancy on future weight development is surprisingly difficult to predict. In the Stockholm pregnancy and weight development study the estimated mean weight retention associated with a pregnancy and estimated 1 year after delivery was 0.5 kg but ranged from -12 to +26 kg. Weight increase during pregnancy was the strongest predictor for sustained weight retention 1 year later. Pre-pregnancy weight did not predict the weight development outcome. The lactation pattern had only a minor influence on weight development. Smoke cessation was an important predictor for sustained weight increase. More weight retention was observed in those women who reported an unfavorable change in lifestyle as regarded eating habits, meal patterns and physical activity. The eventual body weight after pregnancy seems to be more determined by the changes associated with that particular pregnancy than with the lifestyle before. PMID- 9403328 TI - Obesity and breast cancer risk. AB - Being overweight appears to be associated with a higher risk of post-menopausal breast cancer in most studies. Although the relative risk of breast cancer related to Quetelet's index is generally weak (range 1.1-1.9 in the major cohort studies), some studies have found that timing of weight gain and body fat distribution could be more significant factors of an increased risk. Conversely, obesity appears to be slightly correlated with a decreased risk of breast cancer in pre-menopausal women. These contrasting effects of excess weight on breast cancer incidence according to menopausal status, and the lack of a strong association between obesity and breast cancer in some studies, could be due to a number of confounding factors. Among these factors, age, country of origin, family history, alcohol consumption, nutrition, and hormonal treatment could account for the differences observed, and are reviewed in the present study. Obesity and central fat distribution are believed to act through endocrine intermediates such as hyperinsulinaemia and steroid hormones. Since obesity is one of the few breast cancer risk factors that can be modified, the influence of weight loss, particularly in women at high risk, deserves to be further investigated. PMID- 9403329 TI - Weight gain at the time of menopause. AB - The menopause is associated with changes in body composition: a decline in bone mineral content, a decrease in collagen synthesis, a loss of lean body mass and an increase in total and abdominal fat mass. Oestrogen deficiency seems to play a role in the menopause-related changes in body composition, but life styles (diet, exercise, smoking habits, alcohol consumption) are also involved. The time course of the decrease in lean mass deserves attention since it could justify specific actions, i.e. exercising or hormonal treatment, early during the perimenopausal period. A decrease in fat-free mass may be responsible for a decrease in energy expenditure favouring weight gain if the calorie intake is not reduced. PMID- 9403330 TI - Weight gain and cardiovascular risk factors in the post-menopausal women. AB - Cardiovascular disease is the leading cause of mortality and morbidity in the post-menopausal woman. The natural menopause does not appear to be an independent risk factor (or a minor one) for coronary heart disease. Obesity, more precisely excessive intra-abdominal fat, is a cardiovascular risk factor especially with regard to the metabolic risk factors associated with this type of obesity. There is a progressive increase in weight gain at the age of menopause but this weight gain is related to ageing independent of whether women are post-menopausal or not, or treated with oestrogens or not. At the same time, there is a central redistribution of fat with a decrease in gluteo--femoral fat and an increase in intra-abdominal fat with an associated muscle mass loss. This trend to central obesity obviously favours an increased cardiovascular risk. With regard to weight gain, these changes in body composition are related to ageing. Different factors (e.g., diet, physical activity, GH secretion, etc.) may be involved. Are these changes related to menopause? Can hormonal replacement therapy prevent them? The results of the studies in this field are not consistent and these questions remain under debate. PMID- 9403332 TI - Sensory neuropeptides: their role in inflammation and wound healing. AB - Vasoactive neuropeptides including substance P and calcitonin gene-related peptide (CGRP) are localised in sensory nerves which innervate blood vessels. These are the major vasoactive neuropeptides released from sensory nerve endings and both have been suggested to have roles in inflammatory and cardiovascular disease. The neuropeptides have potent effects on microvascular tone and permeability, which are seen soon after release from perivascular nerves. There is also evidence that neuropeptides can affect various activities of inflammatory cells and that sensory nerves play a role in the recovery of the healthy microcirculation during wound healing phases. This review concentrates on evidence that the neuropeptides substance P, acting via tachykinin NK1 and NK2 receptors, and CGRP, acting via CGRP1 receptors, play a pro-inflammatory role in disease and a beneficial role in wound healing. In addition, results from clinical trials of recently developed neuropeptide antagonists are discussed. PMID- 9403331 TI - Vascular leak syndrome: a side effect of immunotherapy. AB - The major dose-limiting toxicity of interleukin-2 (IL-2) and of immunotoxin (IT) therapies is vascular leak syndrome (VLS). VLS is characterized by an increase in vascular permeability accompanied by extravasation of fluids and proteins resulting in interstitial edema and organ failure. Manifestations of VLS include fluid retention, increase in body weight, peripheral edema, pleural and pericardial effusions, ascites, anasarca and, in severe form, signs of pulmonary and cardiovascular failure. Symptoms are highly variable among patients and the causes are poorly understood. The pathogenesis of endothelial cell (EC) damage is complex and can involve activation or damage to ECs and leukocytes, release of cytokines and of inflammatory mediators, alteration in cell-cell and cell-matrix adhesion and in cytoskeleton function. VLS restricts the doses of IL-2 and of ITs which can be administered to humans and, in some cases, necessitates the cessation of therapy. This review discusses the diversity of clinical manifestation, possible mechanisms and therapeutic modalities for VLS induced by IL-2 and ITs. PMID- 9403333 TI - Prevention of ultraviolet radiation-induced suppression of accessory cell function of Langerhans cells by Aloe vera gel components. AB - The active components of Aloe vera gel that can prevent ultraviolet B (UVB) induced suppression of accessory cell function of Langerhans cells (LC) were purified by activity-guided sequential fractionation followed by in vitro functional assay. The functional assay was based on the fact that exposure of freshly isolated murine epidermal cells (EC) to UVB radiation resulted in impairment of accessory cell function of LC, as measured by their ability to support anti-CD3 monoclonal antibody (mAb)-primed T-cell mitogenesis. This UVB suppressed LC accessory cell function was prevented by addition of partially purified Aloe gel components to cultures of UVB-irradiated EC. The Aloe gel components appeared to prevent events occurring within the first 24 h after UVB irradiation that lead to the impairment of accessory cell function. The Aloe gel components did not cause proliferation of anti-CD3 mAb-primed T-cells, nor did induce proliferation of normal EC. The activity-guided final purification of Aloe gel components resulted in the isolation of two components. Both of the components were small molecular weight (MW) substances with an apparent MW of less than 1,000 Da but different from each other in net charge characteristics at pH 7.4. These results suggest that Aloe vera gel contains at least two small molecular weight immunomodulators that may prevent UVB-induced immune suppression in the skin. PMID- 9403334 TI - Inhaled corticosteroids and beta-agonists inhibit oxidant production by bronchoalveolar lavage cells from normal volunteers in vivo. AB - To study the anti-inflammatory mechanisms of inhaled corticosteroids and beta agonist therapies, we evaluated basal and stimulus-induced superoxide production by human airway inflammatory cells obtained by bronchoalveolar lavage from normal volunteers before and after 3 weeks of an inhaled corticosteroid (flunisolide) and beta-agonist (metaproterenol). Assay of superoxide production by the bronchoalveolar lavage cells was performed in the presence of media alone or media containing phorbol ester by optical density determination of reduced ferricytochrome c at 550 nm. Interleukin-1 beta released from unstimulated cells and cells stimulated with lipopolysaccharide was quantitated by enzyme immunoassay. Interestingly, phorbol ester-stimulated superoxide production was strikingly inhibited (P < 0.05) by inhaled therapies, while stimulus induced Interleukin-1 beta production was not significantly affected (P = 0.12). Suppression of oxidant production by airway inflammatory cells may be a major mechanism for the beneficial anti-inflammatory effects of inhaled corticosteroids and beta-agonists. PMID- 9403335 TI - Reduction of disease causative T-cells in experimental autoimmune disease models by a new antirheumatic drug, TAK-603. AB - We investigated the mode of action of a new quinoline derivative, TAK-603 (ethyl 4-(3,4-dimethoxyphenyl)-6,7-dimethoxy-2-(1,2,4-triazol-1-ylmeth yl) quinoline-3 carboxylate), in adjuvant arthritis (AA), a model of rheumatoid arthritis. AA rat splenocytes transferred the arthritis to normal syngeneic rats upon inoculation, but the cells from AA rats treated with TAK-603 (6.25 mg/kg/day) caused only mild arthritis with significantly less foot pad swelling and a lower arthritis score. An effect of TAK-603 in the induction phase of AA was suggested. TAK-603 had little effect on CD4+ and CD8+ T-cell populations in the AA rat splenocytes. We therefore estimated the frequency of T-cells which are reactive to the so-called disease causative antigen using a limiting dilution assay (LDA). The ratio of T cells responsive to PPD, which increased in AA rat splenocytes with the severity of the arthritis, was reduced in AA rats treated with TAK-603. Furthermore, the ratio of MBP (myelin basic protein)-reactive T-cells, which were generated in experimental allergic encephalomyelitis (EAE) rats, were also reduced by TAK-603 administration. These data suggest that TAK-603 acts on the immune system and reduces the number of cells reactive to the relevant antigen. PMID- 9403336 TI - Monophosphoryl lipid A stimulated up-regulation of nitric oxide synthase and nitric oxide release by human monocytes in vitro. AB - Monophosphoryl lipid A (MPL) is a derivative of lipopolysaccharide (LPS) with reduced toxicity which has been shown to modulate various immune functions in monocytes. We examined whether human monocytes can be stimulated to produce nitric oxide (NO) and its catalytic enzyme nitric oxide synthase (NOS). Monocytes were stimulated with LPS or MPL and both NOS and NO (as nitrite) production were measured. MPL at high doses (> 100 micrograms/ml) stimulated monocytes to release NO that was significantly greater than both the control and LPS-treated monocytes (p < 0.05). NO release by control cells and the LPS treated cells was not significantly different. Both arginase and N-monomethyl arginine (NMLA) inhibited the MPL stimulated release of NO (p < 0.01). MPL significantly increased inducible NOS (iNOS) expression as measured by both fluorescent microscopy and flow cytometry (p < 0.05). Similarly, both soluble NOS (sNOS) and particulate NOS (pNOS) activity were significantly up-regulated by MPL (p < 0.05). Significant correlations were found between pNOS expression and sNOS release (r = 0.72, p < 0.0001) and between 12 h NO release and sNOS production (r = 0.44, p < 0.005). These experiments confirm that human monocytes can be stimulated with MPL to produce NO in vitro and suggest that up-regulation of pNOS does not preclude NO release. PMID- 9403337 TI - Superoxide generation by monocytes following infection with human cytomegalovirus. AB - A significant level of superoxide (O2-) generation was observed in a U937-derived subclone following infection with human cytomegalovirus (HCMV). Although there were no detectable levels of viral mRNA and/or protein expression, HCMV DNA content transiently increased immediately before O2- generation. Similarly, O2- generation was also observed in peripheral blood monocytes derived from healthy donors. PMID- 9403338 TI - Immunosuppressive activity of 13-cis-retinoic acid in rats: aspects of pharmacokinetics and pharmacodynamics. AB - Pharmacokinetics and pharmacodynamics of 13-cis-retinoic acid (13-cRA) administered at doses that suppress experimental autoimmune encephalomyelitis (EAE) have been investigated in rats. Serum concentration of the drug measured following oral administration of 37 mg/kg/12 h reached a peak of 1.8 x 10(-5) M in 2 h and linearly declined to 7.8 x 10(-7) M at hour 12. When spleen cells (SC) collected from 13-cRA-administered animals were cultured in vitro, their proliferative response to the T-cell mitogen concanavalin A (ConA) was suppressed and this effect was dependent on in vivo serum concentrations of the drug. In addition, in vitro exposure of antigen-specific T-cell lines to 13-cRA concentrations equivalent to those observed in vivo caused a dose-dependent suppression of the proliferation induced by the antigen as well as by T-cell mitogens. On a molar basis, 13-cRA showed a stronger in vitro immunosuppressive activity than two immunosuppressive agents used in human therapy, cyclosporin A and 6-mercaptopurin. PMID- 9403339 TI - Thymotrophic effect of ether lipid 1-O-octadecyl-2-O-methoxy-rac-glicero-3 phosphocholine in the mouse. AB - 1-O-octadecyl-2-O-methoxy-rac-glicero-3-phosphocholine (ET-18-OCH3) is a synthetic derivate of 2-lysophosphatidyl-choline, endowed with some immunomodulatory and anticancer effects. In the present work we report that a chronic (1 microgram/g b.w. for 2 weeks) or acute single dose injection of ET-18 OCH3 produced a recovery of thymus weight and thymocytes cellularity in two different strains of mice, C57BL6 and Swiss mice, undergoing thymus age-dependent involution. This effect was significant when the thymus weight was reduced at 50% and it was without effect on thymus lacking age dependent involution, such as young mice. The ability of ET-18-OCH3 to produce thymus weight and thymocyte cellularity recovery was also demonstrated in adult mice showing hypotrophy of thymus induced by chronic corticosterone treatment, suggesting that this compound could be effective against thymus hypotrophy induced by external stimuli. This thymotrophic effect of ET-18-OCH3 was not dependent on direct action on thymocyte proliferation, but probably it was dependent on its action on thymic epithelial cells to produce hormone thymulin, which level was found significantly increased in the plasma. These results provide further immunomodulatory propriety of ET-18 OCH3 and open the possibility to use this compound to counteract thymus hypotrophy. PMID- 9403340 TI - A dimeric form of soluble recombinant sheep LFA-3(CD58) inhibits human T-cell proliferation by generating regulatory T cells. AB - We recently observed that the soluble recombinant from of sheep LFA-3, termed sLFA-3 is biologically active as determined by E-rosette inhibition and inhibition of human T-cell proliferation in response to the recall antigen. In the present study, we examined the immunosuppressive properties of a derivative of sLFA-3, a dimeric form of the first domain (D1) of sLFA-3, named sD1Hcys dimer which was made by oxidative binding of the two D1 molecules through disulfide bonds formed between the SH side chains of a cysteine which was added to the C terminal of the D1 domain. By investigating the suppressive properties of the sD1Hcys dimer, we obtained evidence that antigen-stimulated T-cell proliferation was inhibited by the suppressor T cell, mainly CD4 + CD45RA - CD45RO + and CD8 + CD45RA - CD45RO + T cells, generated by incubating PBLs with a low dose (0.5 microgram/ml) of sD1Hcys dimer in the presence of a low dose of IL-2 and GM-CSF. Flow cytometric analysis showed that the expression of some surface molecules on T cells were modulated by a high dose (5 micrograms/ml) of sD1Hcys dimer such as downregulation of CD3 and upregulation of IL-2R, but were not modulated by a low dose (0.5 microgram/ml) of the sD1Hcys dimer. These findings suggest that the sD1Hcys dimer exerts its suppressive effects on the antigen-induced proliferation assay by generating suppressor T cells. The sD1Hcys dimer might therefore have potential as an immunotherapeutic agent to inhibit and/or anergize antigen specific T-cell responses. PMID- 9403341 TI - Inhibition of contact hypersensitivity with different analogs of benzoporphyrin derivative. AB - Four structural analogs of benzoporphyrin derivative (BPD), a potent anti-tumor photosensitizer, were evaluated for their capacity to influence the immunologically-mediated contact hypersensitivity (CHS) response against the hapten 2,4-dinitrofluorobenzene (DNFB). Immunocompetent hairless strain mice received BPD monoacid ring A (BPD-MA, verteporfin) and returned to normal housing conditions or treated with 690 nm red light (transcutaneous photodynamic therapy, PDT). Unexpectedly, we found that mice given BPD-MA exhibited significantly reduced CHS ear swelling responses to DNFB upon antigenic challenge, whether or not they had been treated with PDT. A significant reduction in the CHS response to DNFB was observed when BPD-MA or PDT was given 48 or 24 h prior to, on the same day, or 24 or 72 h after DNFB sensitization. However, the magnitude of the CHS response was unaffected if these treatments were given 96 h after DNFB sensitization, 24 h before challenge with DNFB. Significantly reduced CHS responses also occurred in Balb/c mice given BPD-MA with or without PDT. Mice given BPD-MA but retained in total darkness throughout the experimental period generated full-fledged ear swelling responses to DNFB indicating that CHS suppression with BPD-MA was light dependent. BPD monoacid ring B (BPD-MB) strongly reduced the CHS response of Balb/c mice kept under ambient light while BPD diacid ring A (BPD-DA) and BPD diacid ring B (BPD-DB) also lowered the CHS response but were less effective than the monoacid forms. Other photosensitizers including Photofrin, tin etiopurpurin, and zinc phthalocyanine did not alter the CHS response of Balb/c mice maintained under ambient light. The ability of different BPD analogs to inhibit the CHS response in mice held under ambient light conditions appears related to the potent photosensitizing activity of these compounds. PMID- 9403343 TI - Augmented antibody-mediated footpad responses in mice treated with an anti-IL-4 monoclonal antibody. AB - The effect of the anti-IL-4 monoclonal antibody 11B11 mAb on antigen-induced footpad responses in mice in which Arthus reaction is involved was investigated. Antigen-induced footpad responses were induced by immunization with hen egg lysozyme (HEL) and by challenge injection of HEL into the footpads on day 20 after the immunization. Five hours after the challenge injection, footpad swelling was measured. 11B11 mAb was daily administered i.p. over a period of 10 days, commencing on the day of immunization with HEL. The results showed that the treatment with 11B11 mAb significantly augmented the footpad swelling. There was an increase in the production of anti-HEL IgG2a antibodies in 11B11 mAb-treated mice, while a decrease in anti-HEL IgG1 and IgG antibody production was observed in the animals. The secretion of IL-4 from lymphoid cells of 11B11 mAb-treated mice was markedly reduced. In contrast, increased IFN-gamma production was observed in these animals. Thus, treatment with anti-IL-4 antibodies appears to be effective in augmenting antibody-mediated in vivo responses in which antigen specific IgG2a antibodies and IFN-gamma produced following the antibody treatment play a role. PMID- 9403342 TI - A purine nucleoside phosphorylase (PNP) inhibitor induces apoptosis via caspase-3 like protease activity in MOLT-4 T cells. AB - Children with congenital homozygous deficiency of purine nucleoside phosphorylase (PNP) have abnormalities in purine metabolism that result in T-cell selective immune deficiency. The mechanism of action for cell death has been attributed to intracellular accumulation of dGTP, a potent inhibitor of ribonucleotide reductase and subsequently DNA synthesis, in thymocytes and T-cells but not B cells. However, the mode of cell death has not been determined to be either necrosis or apoptosis. To examine the involvement of apoptosis in T-cells following PNP inhibition, MOLT-4 cells, a human T cell leukemia cell line, were co-treated with the PNP inhibitor, CI-1000 (2-amino 3,5-dihydro-7-(3 thienylmethyl)-4H-pyrrolo[3,2-d]-pyrimidin-4-one HCl), and 2'-deoxyguanosine (dGuo) which resulted in a concentration-dependent loss of cell viability (trypan blue) and inhibition of tritiated thymidine ([3H]-TdR) uptake. Staining of cells with the DNA dye Hoechst 33,258 showed nuclear morphology characteristic of apoptosis. Western blots (24 h lysates) were probed with antibodies against several proteins implicated in apoptosis. Anti-PARP revealed the presence of an 85 kD PARP breakdown product while, anti-alpha-spectrin revealed the accumulation a 120 kD breakdown product, both suggestive of CPP32 cleavage (caspase-3; an ICE like cysteine protease). Western blots also detected the loss of the intact 32 kD caspase-3 isoform, a biochemical event associated with caspase-3 activation. Corresponding fluorometric activity assays detected a marked increase in caspase 3-like activity using the substrate Ac-DEVD-MCA. Lastly, a pan caspase inhibitor (Z-D-DCB) and 2'-deoxycytidine (dCyd), which is known to prevent dGTP accumulation following PNP inhibition, were able to prevent cell death and all indicators of caspase-3-like activity in MOLT-4 cells co-treated with dGuo and CI 1000. In summary, we provided several lines of evidence for the role of apoptosis and the contribution of caspase-3-like proteases in T-cell death following PNP inhibition. PMID- 9403344 TI - Effects of aprotinin on the kallikrein-kinin system in type I diabetes (insulitis). AB - Sub-diabetogenic doses of streptozotocin (STZ) produce insulitis, beta cell destruction and diabetes in mice. Since kinin have been proposed as an inflammatory mediator in several diseases, we decided to evaluate the role of the kallikrein-kinin system in the evolution of insulitis. Male C 57 BL/KsJ mdb mice were injected with STZ (40 mg/kg) for 5 consecutive d. Aprotinin (4000 KIU/d) was injected simultaneously with STZ during 10 d. Plasma and urine samples collected on day 15 were assayed for glucose concentration and proteins, nitrites and kallikrein. Diabetic mice showed hyperglycemia and increased diuresis, marked proteinuria, nitrites and kallikrein. Administration of aprotinin, a potent tissue kallikrein inhibitor, to STZ mice, reduced the hyperglycemia and the altered renal function of the diabetic mice to level no different from normal mice. The present studies are consistent with the hypothesis that the over production of tissue kallikrein in insulitis could be controlled by the effect of aprotinin. PMID- 9403345 TI - Potentiation and inhibition of fMLP-activated exocytosis in neutrophils by exogenous nitric oxide. AB - Exogenous nitric oxide (NO), not derived from NO-donors, but applied directly, could enhance exocytosis of rabbit peritoneal neutrophils induced by suboptimal concentrations of the chemotactic peptide fMLP. The enhancement was maximal at 30 microM NO. Higher concentrations of NO strongly inhibited fMLP-induced exocytosis. The potentiation of fMLP-induced exocytosis by NO could not be reversed by the inhibitors of guanosine-3',5'-cyclic monophosphate (cGMP) accumulation, LY-83583 and methylene blue, or the antagonists of cGMP-dependent protein kinase, Rp-8-pCPT-cGMPS and Rp-8-Br-cGMPS. The concentration of NO needed to enhance fMLP-induced exocytosis was much higher than the concentration leading to an increase in intracellular cGMP levels. These observations suggest that the enhancement of exocytosis by NO is not likely to be mediated by cGMP. At the concentration which inhibited fMLP-induced exocytosis, NO reduced the intracellular level of glutathione. Since it is known that inactivation of intracellular sulfhydryl groups causes complete inhibition of the exocytotic response, it seems evident that the very strong inhibition of exocytosis by high NO concentrations is due to the reaction of NO with glutathione or with other sulfhydryl group-containing targets. PMID- 9403347 TI - Predictions of serum vancomycin concentrations by means of six different equations for calculation of creatinine clearance. AB - Serum vancomycin concentrations were predicted in 59 patients by means of the PKS programme using six different equations to calculate creatinine clearance. These equations are (i) Chiou & Hsu equation, (ii) Cockroft-Gault lean body-weight equation, (iii) Cockroft-Gault total body-weight equation, (iv) Jelliffe equation, (v) Jelliffe & Jelliffe equation, and (vi) Siersback-Neilsen equation. The predictions were made by using (i) the implemented population pharmacokinetic parameters (IPPP), and (ii) the Bayesian method (BM). The results show that (i) if IPPP can lead to under-predictions, and Chiou & Hsu equation is most suited for predictions, and (ii) if IPPP can lead to over-predictions, the Cockroft Gault total body-weight equation is the most suited for predictions. PMID- 9403346 TI - In vivo cimetidine immunomodulatory effects on the cutaneous reaction to oxazolone in the chicken. AB - The effect of the histamine H2-receptor antagonist, cimetidine, on the cutaneous Arthus-like hypersensitivity to oxazolone elicited injecting subcutaneously oxazolone conjugated to egg-albumin (EA-OX) has been examined in the chicken. Cimetidine had opposite effects on the cutaneous reaction to oxazolone in relation to a different immunization schedule. Cimetidine enhanced the cutaneous reaction to oxazolone obtained immunizing chickens with oxazolone dissolved in ethanol (Eth-OX); instead cimetidine inhibited the cutaneous reaction obtained in chickens immunized with oxazolone dissolved in complete Freund adjuvant (CFA-OX). Optimum enhancement of the cutaneous arthus-like reaction to oxazolone occurred when cimetidine was given for three consecutive days starting at the immunization. The enhancing effect was absent in neonatally bursectomized chickens. Moreover, cimetidine stimulated bursal cell proliferation at day 1 after sensitization. The study of the immunoglobulin class of oxazolone antibodies produced in the immunized chickens demonstrated that cimetidine stimulated the IgM oxazolone antibody synthesis in Eth-OX immunized chickens and inhibited the IgY oxazolone antibody production in Eth-OX and total oxazolone antibody production in CFA-OX immunized chickens. The relationship between increased IgM oxazolone antibody synthesis and enhancement of the cutaneous Arthus reaction is discussed. The role of IgM antibodies in the pathogenesis of Arthus reaction in the chicken is hypothesized. PMID- 9403348 TI - Efficacy of alpha,beta-arteether in acute uncomplicated P. falciparum malaria. AB - A phase-III clinical trial was conducted in 50 patients (42M + 8F) with acute uncomplicated falciparum malaria from Delhi during the period of September to November 1995. Their mean age was 27.2 years, and the mean parasitaemia on day 0 was 0.65%. Patients were hospitalized and treated with a new ethyl derivative of artemisinin developed at CDRI called alpha, beta-arteether, at the dosage of 150 mg l/M for three consecutive days. Peripheral smears were examined every day for 4 days and then weekly up to 28 days. The results of the study showed that the mean parasite and fever clearance times were respectively 19.94 +/- 6.87 and 37.81 +/- 21.67 hours. Within 48 h, 70% of the cases became afebrile and the peripheral smear was negative in 100% of the cases. The drug was well tolerated. Three cases (6%) had recrudescence within 28 days. It is concluded that alpha, beta-arteether is a safe, effective and rapidly acting antimalarial. PMID- 9403349 TI - Evaluation of clinical patterns in ulcerative colitis: a long-term follow-up. AB - The aim of this prospective research was to compare, in a seven-year follow-up, the clinical outcome of ulcerative pancolitis with that of non-progressive ulcerative colitis. The activity of the disease was evaluated by a Clinical Activity Index and an Endoscopic Index. Of 112 cases of ulcerative colitis observed, 95 showed no change in extent and were studied as examples of non progressive UC, and in this group the extension of the disease was: pancolitis in 19%, left-sided colitis in 39%, proctosigmoiditis in 17% and proctitis in 25%. A colectomy had to be performed in 5%. None of the enrolled cases developed a cancer during the follow-up. The patients with ulcerative pancolitis or left sided colitis were treated with 5-ASA 1.6 g/day in a delayed-release formulation, while the cases with proctosigmoiditis or proctitis were treated with 5-ASA enemas 4 g/day. The cases with more than one relapse/year were 39%. The proportion of patients with only one relapse/year was 53%. The patients with steady remission for all the seven years of the trial were only 8%, but with a statistically significant difference between the groups with initial diagnosis of proctosigmoiditis or proctitis and the group with initial diagnosis of pancolitis or left-sided colitis (12% versus 5%). Among the cases with continuous remission, 37% showed colonic alterations, with an endoscopic score higher than 4 but a clinical score less than 6. Side-effects were observed in 6% patients but without treatment withdrawal. Non-progressive ulcerative colitis throughout the colon has a relatively good prognosis which seems to be independent of the location of the disease, even if we have found a statistically significant higher percentage of cases with steady remission among the patients with more distal disease. PMID- 9403350 TI - Synthesis and disposition of 14C-labelled 81/470, a new anthelminthic agent in rats. AB - CDRI compound 81/470 (AH) is a new broad-spectrum anthelminthic agent under development for clinical and veterinary application. We herewith report the synthesis of 14C-labelled AH, and a study of the tissue distribution and excretion of radioactivity after administering a single 1 mg/kg p.o. or i.v. dose in young male rats. After oral administration of the dose, percent radioactivity recovered in 24-h urine, faeces and tissues were 17.9, 59.7 and 22.9 respectively. The levels were below detection limit in brain and gonads up to 24 h. In bile-duct-cannulated rats, the majority (37.8 +/- 2.8 and 43.8 +/- 6.4) of the radioactivity was excreted in the bile within 24 h of p.o. and i.v. administration, respectively. After an oral dose (1 mg/kg), the urinary excretion of radioactivity in rats was found to be approximately one-half (21 +/- 5.7, 18.3 +/- 2.1) of that obtained by i.v. administration of an equal dose (40.2 +/- 3.1 and 35 +/- 1.3), in bile-duct-intact and cannulated rats respectively. PMID- 9403351 TI - Natural killer (NK) cell assay within bladder mucosa of patients bearing transitional cell carcinoma (TCC) after interferon (IFN) therapy: an immunohistochemical and ultrastructural study. AB - The authors studied the number and the ultrastructural evidence of NK cell activation in the non-involved urothelium in patients with transitional cell carcinoma (TCC) of the urinary bladder, before and after transurethral resection (TUR) and interferon (IFN) therapy. Eight male patients, free of recurrence 1 year after TUR and IFN-a2b therapy, were studied. Each patient received 22 instillations of 50MU of IFN-a2b over a period of 1 year. Two specimens from the non-involved urothelium, one adjacent to and another away from the tumour, were obtained before and after therapy, for immunohistochemical and ultrastructural studies. The number of NK cells was evaluated immunohistochemically in paraffin sections with the CD57 monoclonal antibody, and their activation was detected by routine electron microscopy processing. Before treatment, few NK cells were randomly found in the lamina propria. At the end of therapy, however, their number increased and NK cells were found to infiltrate the urothelium, a finding that was not observed before treatment. The number of NK cells did not correlate with the degree of the inflammatory infiltrate of the mucosa. Moreover, the ultrastructural study revealed activation of NK cells with enhanced cytolytic activity. IFN therapy increases the number and promotes the activation of NK cells within the bladder mucosa. This finding could be of clinical significance in the prevention of tumour recurrence, given that NK cells enhance the immunological defense mechanisms of the bladder. PMID- 9403352 TI - Intracerebroventricular self-injection of beta-endorphin by rats in response to intermittent electrical shocks. AB - Rats were taught to self-administer beta-endorphin (0.1 microgram or 0.5 microgram/microliter) or NaCl 9% by pressing a lever that activated a pump connected with a cannula implanted in the lateral cerebral ventricle (icv). The pump delivered 1 microliter at each lever pressing. After a training period of 2 weeks, the self-injection behaviour was studied before, during and after nociceptive stimulation. In response to a nociceptive stimulation, 4 rats increased their self-injection of beta-endorphin at 0.5 microgram/microliter per injection. This effect is specific for beta-endorphin since under identical conditions 6 rats did not increase the injection of isotonic NaCl saline solution. Another 6 rats did not increase their self-injection during nociceptive electrical stimulation and the post-nociceptive period when the dose of beta endorphin was 0.1 microgram/microliter per injection. However these animals self administered significantly higher levels of beta-endorphin during the pre-control period. Also studied were the acute effects of 10 micrograms of icv beta endorphin on tail-flick latency in sec. The acute administration of 10 micrograms of beta-endorphin induced a long-lasting analgesia. The results show that the rats increased beta-endorphin self-administration during the pre-control period when the dose was 0.1 microgram/microliter and during the nociceptive stimulation period when the dose was 0.5 microgram/microliter. In the former case the self administration had the profile of a voluntary doping drug intake, in the latter case the profile was that of an antalgic self-medication. PMID- 9403353 TI - Effect of flurenizide on adaptive reactions in patients with chronic obstructive pulmonary disease. AB - A total of 75 patients with chronic obstructive pulmonary disease (COPD) was investigated, 50 of which received Flurenizide in addition to traditional therapy. The remaining 25 received traditional therapy only and constituted the control group. Clinical improvement was observed in both groups, but this was attained in different ways. This depended on the type of the adaptive reactions at the beginning of exacerbation and the transition from one reaction to the other. In the basic group that received Flurenizide, clinical improvement was attained by a transition from physical conditions of defective adaptation (DA) or reaction of orientation (RO) into positive reactions of slight (RSA) or elevated (REA) activation in 72% of patients. Negative transition was not observed, and 14% of the patients remained in the DA condition. In the control group, a transition from DA into RO was observed in 27% of the patients, while 73% remained in DA without any positive adaptive reactions being definable. A significant improvement in the parameters of immunological and lung function tests was observed only in the group of patients who received Flurenizide. The highest levels of the above-mentioned parameters were observed in patients whose adaptive reactions had transited into the RSA or REA condition. We suggest that the reason for this positive dynamics was induced by the immunostimulating effect of Flurenizide. PMID- 9403354 TI - Evolution of the dopaminergic system and its relationships with the psychopathology of pleasure. AB - This paper summarizes the fundamental steps in the evolution of the dopaminergic system. A rudimentary dopaminergic system is present in primitive creatures, already able to select information processing, modulate "emotional" behaviours and react to perturbations in environmental conditions. Pharmacological manipulations of the dopaminergic transmission are able to modify basic behaviours present in all animals from fishes to lizards to mammals. The ability to put the organism in motion and the hedonic capacity of giving pleasure, would justify the conservation through evolution of such a neuronal system. The fact however that the dopaminergic system has remained identical for the last several centuries, while many external conditions which interfere with its physiology have dramatically changed, may contribute to explain the transition from the original vital advantages of the dopaminergic system to its crucial role in the psychopathology of pleasure. PMID- 9403355 TI - Immediate-early genes expression in spinal cord as related to acute noxious stimulus. AB - During the "central sensitization" phenomenon, noxious stimuli lead to expression of IEGs (c-fos, c-jun, krox-24); their proteic products have been postulated to convert short-term stimulations into long-lasting responses in dorsal-horn neurons. The aim of this study was to verify if analgesic drugs, such as morphine and ketorolac, may affect the c-fos protooncogene expression by using a method highly sensitive and specific, based on transformation of activated c-fos specific mRNA in cDNA (reverse transcription), its amplification (PCR) and final visualization by electrophoresis on agarose gel. Male Wistar rats were submitted to various stimuli in order to assess which procedure resulted in genic derepression; monolateral sciatic nerve ligature appeared to be the most effective. When the animals were pretreated with morphine or ketorolac and subsequently exposed to the monolateral sciatic nerve ligature, or treated with ketorolac immediately after the same painful stimulus, we found that only pretreatment with morphine completely blocked c-fos depression. Our results confirm that pretreatment with opioids is able to prevent IEGs derepression and the central sensitization phenomenon. PMID- 9403356 TI - Central neuronal changes in recurrent visceral pain. AB - Repeated colics from urinary calculosis produce referred muscle hyperalgesia whose extent is proportional to the number of colics. The pathophysiology of this hyperalgesia was investigated in electrophysiological studies (spinal cord recordings) on an animal model of artificial ureteral calculosis. Stone-implanted rats show both visceral pain episodes and muscle hyperalgesia of the ipsilateral lumbar musculature; the extent of hyperalgesia is a function of the number of episodes. In the dorsal horn of stone-rats compared to controls the following were found: a) significantly higher percentages of neurons driven by stimulation of the hyperalgesic muscle, of spontaneously active cells with muscle input and of neurons with muscle input with a low mechanical threshold of activation, and b) a significantly higher frequency of background activity of spontaneously active cells with muscle input. These findings were proportional in extent to the number of visceral episodes presented by the rats before recordings; in cases of an extremely high number (> 50), several neurons also displayed abnormal activity, i.e. permanent short rhythmic bursts. These changes reflect a state of central sensitization and are probably due to the abnormal inflow from the affected ureter which facilitates the central effect of muscular input, thus accounting for the referred hyperalgesia. The degree of sensitization appears to be a function of the repetition of the visceral afferent barrage. PMID- 9403357 TI - Decentralization monoamine super-sensitivity of migraine and opiate abstinence: common features and different target mechanisms? AB - The similarity between opiate withdrawal and migraine (M) has been confirmed regarding increased monoamine sensitivity at the neuromuscular junction of the hand's dorsal vein as well as at the neuraxis where dopamine (DA) supersensitivity was observed. Similarities also included an increase in cAMP levels as a precocious sign in both M and opiate withdrawal. Particular attention has been devoted to the time-course of monoamine supersensitivity in M and in abstinence. It has been found that the maximum level of super-sensitivity occurs in M at the end of the M attack, whereas the maximum super-sensitivity is present at the very beginning of opiate abstinence. The inverse time-course of this phenomenon suggests that it could play some pathophysiological role in inducing the end of the M attack. Conversely, it can represent the expected transient result of a pharmacological denervation which ought to result in a supersensitivity of opioid-dependent neuron during withdrawal. In M, the super sensitivity is wider, indeed, it involves more receptor types. This could be an indirect proof of the involvement of inhibitory pathways other than the opioidergic one. PMID- 9403358 TI - Hypersensitivity to meta-chlorophenylpiperazine (mCPP) in migraine and drug withdrawal. AB - Administration of a bolus dose of mCPP, a 5-HT2C receptor agonist, to rats provokes endocrine and behavioural effects that are reminiscent of some of the symptoms of human depression. Rats exposed to chronic mild stress (which is also a key factor in the precipitation of human depression) were hypersensitive to mCPP, whilst chronic treatment with antidepressant serotonin re-uptake inhibitors suppressed the responsiveness to mCPP. Similarities also exist with respect to withdrawal reactions following chronic alcohol or benzodiazepine abuse. In humans, a bolus dose of mCPP can cause alcohol craving (in abstinent alcoholics) and migraine (in susceptible persons), suggesting that there is a 5-HT2C receptor hyperresponsiveness in these conditions also. It is hypothesized that chronic treatment with SSRI's can prevent migraine attacks and drug craving. PMID- 9403359 TI - The way to serotonergic use and abuse in migraine. AB - 5-HT is currently indicated to play a role in migraine (M). Previously evidenced 5-HT supersensitivity which characterizes M is insufficient to compensate for a possible deficit in 5-HT bioavailability. Inducing a further up-regulation of 5 HT receptor can yield improvement of M syndrome. Chronic treatments of methysergide and propranolol, drugs exerting antagonist action at 5-HT receptors, induced a significant amelioration in 256M sufferers. On the contrary, chronic treatments of ergotamine and sumatriptan, both provided with a 5-HT1 agonist activity, induced a worsening of M in 134 M sufferers. The M worsening was paralleled by an increase in consumption of analgesic drugs. Discussion concerns the effects of the chronically given 5-HT agonists and antagonists as well as the possible receptor mechanism underlying "craving for serotonin" in severe M. The increase of 5-HT supersensitivity evidenced at the end of M attacks is also discussed and its role in determining the interruption of the attack is here suggested. PMID- 9403360 TI - Inheritable and acquired hyperalgesia associated to abnormal opioid receptor set up seem to act as opiate addiction preventors. AB - Hyperalgesia is known to depend on neuroplastic changes chiefly represented by long-term potentiation. These phenomena are proved to depend on excitatory amino acids (EAAs) action at the level of NMDA receptors. This action is known to be related to nitric oxide (NO) release. We found a visceral/vascular hyperalgesia state in migraine (M) sufferers as well as an inheritable systemic hyperalgesia in healthy subjects who are first-degree consanguineous with M sufferers; this type was labelled 'third hyperalgesia'. We discovered that a hyper-increase of plasma L-citrulline, equimolar co-product in the synthesis of NO, characterizes both M sufferers and their first-degree relatives who are exempt from primary headache. A similar pattern never occurred in healthy subjects having both a personal and family history negative for primary headache. We conclude that both 'third hyperalgesia' and a pattern of NO synthase (NOS) hyperactivity seems to be inheritable and can constitute, at least in part, a ground for developing headache. Morphine, proved to be unable to relieve M attack, was given in low doses that caused pain and side-effects in M sufferers only. This outcome seemingly indicates that M sufferers are characterized by a set-up of opioid receptor subtypes different from that of healthy headache-exempts. Following a period of morphine addiction and a withdrawal period, 65.07% of a group of 63 opiate addicts developed M syndrome. All these subjects were first-degree consanguineous relatives of primary headache sufferers. Discussion topics concern the activity of morphine in NO release and the role of NO in sensory transmission of both controls and hyperalgesia sufferers. It is suggested that the inheritable couple consisting of hyperalgesia and NOS hyperactivity can play some role in setting off the pain occurring following morphine in M sufferers. PMID- 9403361 TI - Bradykinin and cerebral circulation: selective non-receptor interaction with histamine and serotonin through the system of nitric oxide. AB - Bradykinin, histamine and serotonin were the most active physiological agonists in increasing jugular levels of nitric oxide when administered into the left common carotid artery of anaesthetized rabbits. Bradykinin potentiated selectively the effects of histamine and serotonin (but not vice versa) on both nitric oxide and carotid blood flow, without involving activation of specific receptors. Since these effects were demonstrated using doses of the three agonists in the order of their physiological plasma concentrations, it was concluded that cerebral circulation in regulated, also within its autoregulatory limits, by bradykinin through selective autocoidal interactions converging on nitric oxide. PMID- 9403362 TI - Bioeffects of a nitric oxide donor in a human preosteoclastic cell line. AB - Nitric oxide is a short-lived free radical produced by different isoforms of the enzyme nitric oxide synthase. It regulates a whole range of functions in the body, but little is known about its effects on bone. Rat osteoclasts and a human preosteoclast cell line (FLG 29.1) have been shown to produce nitric oxide and to express nitric oxide synthases. In the present study we investigated the role of a nitric oxide donor, 3-morpholinosydnonimine hydrochloride, on the FLG 29.1 cells. 3-Morpholinosydnonimine hydrochloride has been shown to significantly increase IL6 production and tartrate resistant acid phosphatase activity in FLG 29.1 cells, indicating a positive modulation of osteoclast differentiation. However FLG 29.1 cell adhesion on osteoblast-like cells was significantly inhibited, suggesting an inhibition of osteoclast motility. All these results confirm the bidirectional effect of nitric oxide whose basal production is necessary in promoting osteoclast differentiation, while at high levels it is effective in inhibiting osteoclast activity. PMID- 9403363 TI - Modulation of excitatory amino acids pathway: a possible therapeutic approach to chronic daily headache associated with analgesic drugs abuse. AB - The use of antagonists of N-methyl-D-aspartate (NMDA) receptors, or the administration of inhibitors of the synthesis or of the release of excitatory amino acids, enables the analgesic drug-dependence associated with chronic daily migraine to be overcome without any physical abstinence sign. Follow-up period indicates that negative modulators of excitatory amino-acid function can induce a stable benefit. The persistent benefit is seemingly due to an inhibitory effect on the process underlying the hyperalgesia state which is a crucial feature of migraine. It can also be suggested that the antagonist activity at NMDA receptor might play a role in very severe non-opioid analgesic drug dependence. PMID- 9403364 TI - Therapy of migraine by modulating dopamine hypersensitivity: its effect on mood and pain. AB - Clinical and pharmacological evidences support the hypothesis of a hypersensitivity of dopamine (DA) receptors in migraine patients. The aim of our single-blind and placebo-controlled study is to evaluate the presence of this hypersensitivity and to desensitize the DA system with apomorphine, a classical DA receptor agonist. We studied six patients suffering from migraine without aura resistant to the prophylactic treatment and with high frequency of attacks. In these patients, we first tested individual sensitivity by a single i.m. low dose of apomorphine to assess the lowest effective dose to be administered s.c. in continuous infusion without side-effects. We used the response of growth hormone (GH) to apomorphine as a marker of DA2-receptor function. Clinical evaluation, blood pressure and heart rate were recorded before placebo or apomorphine administration and monitored every 15 min for 1 h after each injection. The treatment consisted of the continuous subcutaneous infusion of apomorphine for three weeks at the dosage previously assessed by the test. No difference in blood pressure and heart rate was found during test and treatment. The treatment was effective in reducing the frequency and the severity of migraine attacks. PMID- 9403365 TI - Involvement of central cholinergic system in antinociception induced by sumatriptan in mouse. AB - The antinociceptive effect of the antimigraine drug sumatriptan was assessed in mice (hot-plate and abdominal constriction tests). Antinociception induced by sumatriptan (10-30 mg kg-1 i.p.) was prevented by the muscarinic antagonist atropine (5 mg kg-1 i.p.), the ACh-depletor hemicolinium-3 (1 microgram per mouse i.c.v.) and the 5-HT1A antagonist NAN-190 (0.5 mg kg-1 i.p.). Naloxone, CGP-35348 and reserpine administered in doses suitable for blocking analgesia respectively induced by morphine, baclofen and clomipramine did not modify sumatriptan antinociception. On the basis of the above findings, we can deduce that sumatriptan was able to induce antinociception by increasing cholinergic neurotransmission through the stimulation of 5-HT1A receptors. PMID- 9403366 TI - Stress, mood disorders and memory in headache. AB - Comorbidity between headache and other disorders such as psychological or memory problems is a topic of increasing scientific interest both for us diagnostic and therapeutic implications but also for pathogenetic advances. A central neurogenic mechanism such as a dysregulation of some neurotransmitter system might underlie not only headache but also other coexistent disorders; findings highlight the role of serotonin pathways. PMID- 9403367 TI - Intermittent but not continuous inescapable footshock stress and intracerebroventricular interleukin-1 similarly affect immune responses and immunocyte beta-endorphin concentrations in the rat. AB - Both CNS- and immunocyte(lymphocytes, splenocytes)-derived beta-endorphin is involved in immune responses to stress. We show in the rat that stress-induced immunodepression (decrease of mitogen-induced lymphocyte proliferation and NK activity) is present only after the administration of a stress paradigm that increases immunocyte-derived beta-endorphin, while this is absent when the concentrations of the opioid are not modified. Interestingly, plasma corticosterone levels were similarly elevated after stresses whether or not they suppress immune responses, thus suggesting a pivotal role of the opioid. The increase of immunocyte beta-endorphin and immunosuppression are similarly present also after the intracerebroventricular administration of interleukin 1, thus suggesting a role for this cytokine in stress responses. The modifications of immunocyte beta-endorphin concentrations and immune responses induced by stress and interleukin 1 are not affected by indomethacin, adrenalectomy or hypophysectomy, whereas they are completely blocked by a CRH antagonist and depletion of the serotoninergic or catecholaminergic systems. In conclusion, our results suggest that immune responses to stress are not uniquely linked to an activation of the HPA axis. PMID- 9403368 TI - Psychological characteristics of juvenile headache: differences between tension headache and migraine. AB - In 235 juveniles equalized for sex and averaging 10 years of age, comparisons were made between 76 with migraine without aura, 79 with episodic tension type H, and 80 normal controls, on the basis of five psychological characteristics and six formal tests. The results allowed the description of definitely different profiles for the two groups of headache sufferers. PMID- 9403369 TI - Diagnosing autism: analyses of data from the Autism Diagnostic Interview. AB - Results from ROC curves of items from two scales, the Autism Diagnostic Interview (ADI) and Autism Diagnostic Interview-Revised (ADI-R), operationalizing DSM-IV criteria for autism are presented for 319 autistic and 113 other subjects from 8 international autism centers. Analyses indicate that multiple items were necessary to attain adequate sensitivity and specificity if samples with varying levels of language were considered separately. Although considering only current behavior was generally sufficient when a combination cutoff and additive model was employed, predictive power was highest when history was taken into account. A single set of criteria, as operationalized by individually structured questions in the ADI/ADI-R, was effective in differentiating autism from mental handicap and language impairment in subjects with a range of chronological ages and developmental levels. PMID- 9403370 TI - Does teaching theory of mind have an effect on the ability to develop conversation in children with autism? AB - The present research examined whether teaching children with autism to pass tasks that assess mental state understanding had any positive effects on communication. Two aspects of communication previously shown to be deficient in children with autism were considered. These are conversational ability, in particular the ability to expand on conversation, and the use of mental state terms in speech. Results showed that no discernible improvement was seen on either measure of communication following mental state teaching. Discussion centers on real versus superficial changes in understanding mental states as a result of teaching. PMID- 9403371 TI - Are deficits in the decoding of affective cues and in mentalizing abilities independent? AB - It has been hypothesized that deficits in theory of mind (ToM) and emotion recognition abilities in subjects with autisticlike disorders are independent. We examined the relationships between deficits in the various social cognitive domains in children with an autistic disorder (N = 20), in children with a pervasive developmental disorder not otherwise specified (PDDNOS) (N = 20), and in psychiatric control (N = 20) and normal children (N = 20). The clinical groups were matched person-to-person on age and verbal IQ. The clinical children were 8 18 years old, the normal children 8-13 years old. The test battery included tasks for the matching and the context recognition of emotional expressions, and a set of first- and second-order ToM tasks. ToM and emotion recognition functioning proved to be better integrated in the non-PDD children than in the PDD children, but also in the PDD children significant correlations were found between ToM and emotion recognition measures. PMID- 9403372 TI - Delay versus deviance in autistic social behavior. AB - The pattern of acquisition of social, communication, and daily living skills was examined for autistic children, compared to retarded and normal controls, by quantifying intradomain scatter on the Vineland Adaptive Behavior Scales. Autistic children were matched to normal children and mentally retarded children on Vineland raw scores; group differences in scatter were examined for each domain of adaptive behavior. Autistic children had significantly more scatter on Communication and Socialization than both control groups. Item analyses showed that the autistic children had particular weaknesses on items reflecting attention to and pragmatic use of language, as well as play and reciprocal social interaction; the autistic children had particular strengths on items reflecting written language and rote language skills, and rule-governed social behavior. The number of items showing consistent group differences, however, was small, suggesting that although autistic development appears sequentially deviant and not merely delayed, individual autistic children derive their scatter from different items, and are a developmentally heterogeneous group. PMID- 9403373 TI - The role of touch in facilitated communication. AB - Imagine that one day a nonverbal autistic child suddenly starts to type messages, such as "I am not retarded," using a computer keyboard while being touched by an assistant. Facilitated communication (FC) appears to create this miracle around the world. To understand how this works, experiments were conducted involving a "telepathy game" using a rod with an attached strain gauge. A force from the assistant, which controlled what was spelled through physical support, was measured. It was thus completely possible for any message to appear to be typed with FC regardless of the autistic child's actual knowledge or language ability. PMID- 9403374 TI - Brief report: motor incoordination in children with Asperger syndrome and learning disabilities. PMID- 9403376 TI - Brief report: interrater reliability of the psychoeducational profile (PEP). PMID- 9403375 TI - Brief report: electroencephalographic paroxysmal activities in the frontal area emerged in middle childhood and during adolescence in a follow-up study of autism. PMID- 9403377 TI - Piracetam: a novel therapy for autism? PMID- 9403378 TI - Sensation Seeking Scales and consumer satisfaction with a substance abuse treatment program. AB - Satisfaction of 119 addicts with an addiction treatment program was measured by an 11 item satisfaction scale. The scale's internal consistency was acceptable (Cronbach's alpha = .75). The total satisfaction score was weakly but significantly correlated with Zuckerman's Sensation Seeking scales: Those with higher scores on the Boredom Susceptibility scale (i.e., those easily bored) reported less satisfaction, whereas those with higher scores on Thrill and Adventure Seeking scale (i.e., risk, adventure, and thrill seekers) reported higher levels of treatment satisfaction. Older patients were more satisfied with the feedback they received from their psychological tests and also with staff's respect for their rights. PMID- 9403379 TI - Readability of self-report clinical outcome measures. AB - Because the validity of data obtained from self-report clinical outcome measures depends upon the ability of the client to comprehend the inventories, readability was assessed for five frequently employed measures: Beck Depression Inventory, Integra Outpatient Tracking Assessment, MOS 36-Item Short-Form Health Survey, Social Adjustment Scale-Self Report, and Symptoms Checklist-90-Revised. The Flesch Reading Ease (RE) formula and a Flesch abstraction formula were applied. The measures are generally shown to be useful for patients with an eighth or ninth grade education, suggesting that outcome researchers must choose only those measures appropriate to the educational background of their clients. PMID- 9403380 TI - MMPI-2 profiles of women differing in sexual abuse history and sexual orientation. AB - Four groups of women (N = 115) self-identified as having histories of childhood sexual abuse or no such histories and self-identified as either heterosexual or lesbian were compared using a questionnaire and the MMPI-2. Subjects ranged in age from 21-60 years with 60% between ages 30-50 years. Results of a Three-Way MANOVA for abuse history and sexual orientation repeated across MMPI-2 clinical scales showed a between-subjects effect for abuse, and within-subjects effects for orientation and abuse. T scores of women with abuse histories were significantly higher than those of women without abuse histories on Hs, D, Pd, Pa, Pt, Sc, and Ma scales of the MMPI-2. Profiles indicated an 8-4 codetype and a Scarlett O'Hara V configuration for the group with abuse history. Heterosexual women obtained significantly higher t scores than did lesbians on the Depression scale. Results show that the MMPI-2 can be used to help detect lesbian as well as heterosexual adults who were sexually molested as children. PMID- 9403381 TI - Combat guilt and its relationship to PTSD symptoms. AB - Guilt regarding combat experiences is often considered an associated symptom of PTSD in military veterans. Little is known, however, about the role combat guilt plays in the development and maintenance of PTSD. Inadequate measurement of combat-related guilt may be one reason for this deficiency in the literature. In the present study, 40 veterans with PTSD completed a novel measure of combat guilt. Items on the scale assessed various types of guilt and shame concerning combat experiences (i.e., survival guilt, guilt over acts of omission and acts of commission, guilt about thoughts/feelings). Guilt was quite prevalent within this sample, and severity of guilt regarding combat was positively correlated with the reexperiencing and avoidance symptoms of PTSD and a general measure of PTSD severity. Implications of these findings and recommendations for the development of measures for combat-related guilt are discussed. PMID- 9403382 TI - Assessment of PTSD symptoms in a community exposed to serial murder. AB - This study examined the presence of PTSD symptoms across time in a community exposed to serial murder. One hundred eighty four subjects (48% response rate) responded to the initial survey while 64 and 30 subjects, respectively, participated in the 9- and 18-month follow-up studies. Results indicated widespread endorsement of PTSD symptoms following the murders. The most severe reactions were found among residents demographically similar to the victims. PTSD symptoms, while not transient, appeared to decrease over time with few subjects still reporting symptoms at 18 months. These data suggest that violent acts such as serial murder can have far reaching psychological consequences for the community and result in vicarious victimization. PMID- 9403383 TI - The brief admission unit in emergency psychiatry. AB - The study evaluates a Brief Admission Unit for clients of an emergency service located within a comprehensive community psychiatric program. Eighty-five clients completed the Brief Symptom inventory and a structured interview. Substance abuse disorder (n = 29) and major depression (n = 24) were the most common Axis I diagnoses, of which 30 subjects had two or more. Sixty subjects had an Axis II diagnosis. Mean duration of admission was 3.9 days, compared with the average in other acute units of 11.5 days. At discharge, half the subjects were rated as moderately to greatly improved and client satisfaction was high. The unit was crucial to the psychiatric emergency service and had a key role in relieving pressure on beds elsewhere within the system. PMID- 9403384 TI - MCMI-III as a treatment outcome measure for psychiatric inpatients. AB - The Million Clinical Multiaxial Inventory-III (MCMI-III) recently was introduced to replace and update the MCMI-II. A sample of 97 psychiatric inpatients were administered the MCMI-III shortly following admission, and again 7-10 days later. Changes in the personality and symptom scales generally paralleled those found in previous work with the MCMI-II, although the mean retest interval was considerably shorter than in the earlier study. However, some differences between the two instruments were observed, confirming the need for ongoing cross validation work on the MCMI-III as an instrument that is distinct from the MCMI II. PMID- 9403385 TI - Stability of psychiatric patients' perceptions of their admission experience. AB - The primary aim of this study was to evaluate the stability (i.e., consistency of patients' responses over time) of newly developed scales to measure the admission experience of psychiatric hospitalization. Eighty-four psychiatric patients involuntarily committed to a crisis stabilization unit participated. All participants were admitted under an emergency petition or ex parte order for a psychiatric evaluation. Patients were interviewed soon after admission (M = 3.33 days, SD = 1.86 days). The test-retest interval was 24-48 hours with most (83.3%) re-evaluated at 24 hours. Overall, the measures showed acceptable levels of stability (r's range from .62 to .72). Factors associated with reliable responses were lower overall psychiatric symptom severity, less severe psychotic symptoms, and mentioning the same person as an influence of perceptions about the admission experience at each assessment point. PMID- 9403386 TI - Utilizing the PK scale of the MMPI-2 to detect posttraumatic stress disorder in college students. AB - This study investigated the utility of the PK scale of the MMPI-2 with college students. Results indicated that the PK scale, when combined with DSM IV criteria, does discriminate between college students who obtain a score of 65 or higher and those who score below 65. PMID- 9403387 TI - Minnesota Multiphasic Personality Inventory profiles of Vietnam combat veterans with posttraumatic stress disorder and their children. AB - Forty children of 28 fathers who are Vietnam veterans with posttraumatic stress disorder (PTSD) completed the Minnesota Multiphasic Personality Inventory. Each of the fathers had at least one elevated clinical scale. Fathers averaged eight elevated clinical scales, and compared to more recent norms, fathers averaged seven elevated clinical scales. Seventy-eight percent of the children had at least one clinically elevated scale (averaging three elevated clinical scales). Compared to contemporary normal adolescents and adults, 65% of children had at least one clinically elevated scale (still averaging three elevated clinical scales). No consistent MMPI profile patterns emerged within or across the two groups. No gender differences were detected among child MMPI profiles. Forty percent of the children reported illegal drug use, and 35% reported behavior problems. Fifteen percent of children reported previous violent behavior. Eighty three percent of the children reported elevated Cook-Medley hostility scores as compared to an age-matched national normative sample. Children with higher PK scores were also significantly more likely to report higher Cook-Medley hostility scores. Forty-five percent of children reported significant elevations on the PTSD/PK subscales. PMID- 9403388 TI - A preliminary evaluation of treatment outcomes at a veterans' hospital's inpatient psychiatry unit. AB - This article describes a preliminary effort to evaluate inpatient psychiatric services at the Carl Vinson DVA Medical Center in Dublin, Georgia. The facility annually treats a large number of veterans for a variety of psychiatric disorders. To determine whether these veterans improved following care, a simple pretest-posttest group design was employed, using the SCL-90-R, to assess psychiatric symptomatology before and after inpatient treatment. Both statistically significant and practically meaningful improvements in symptomatology were evident at discharge. While the research design does not permit causal inferences, low-cost evaluations such as this one simply demonstrating that patients get better are important first steps in empirically determining the efficacy of inpatient psychiatric services, and represent one means of demonstrating accountable practice. PMID- 9403389 TI - Interpersonal violence and its correlates in Vietnam veterans with chronic posttraumatic stress disorder. AB - Two studies were conducted to investigate interpersonal violence in Vietnam veterans with posttraumatic stress disorder (PTSD). In study one, combat veterans with PTSD reported significantly greater occurrence of violent behaviors over the past year (22 acts) versus combat veterans without PTSD (.2 acts). Combat exposure had an independent positive association with interpersonal violence. In study two, variables related to current interpersonal violent behavior in 118 PTSD combat veterans were evaluated. In rank order of importance, lower socioeconomic status, increased aggressive responding and increased PTSD severity were related to interpersonal violence. These results suggest that combat veterans with PTSD exhibit greater interpersonal violence than combat veterans without PTSD, and that there are multiple factors in this population which determine violent behavior. PMID- 9403390 TI - Relationship between scores on anger measures and PTSD symptomatology, employment, and compensation-seeking status in combat veterans. AB - The interrelationship between the theoretically related constructs of anger and posttraumatic stress disorder (PTSD) symptoms was examined in a group of 42 combat veterans with PTSD using a multimeasure assessment strategy. Scores on several anger measures were found to be quite high in this sample and were significantly correlated with PTSD symptomatology. Furthermore, anger measures were found to be related to employment status independent of PTSD severity, but were not related to disability compensation-seeking status. Clinicians are advised to be aware of the potential implications for physical health and interpersonal functioning, and to incorporate anger management strategies into treatment plans for this population. PMID- 9403391 TI - The MMPI-2 GM and GF Scales as measures of psychological well-being. AB - The MMPI-2, Symptom Checklist (SCL-90-R), and Tennessee Self-Concept Scales (TSCS) scores were analyzed for 117 male and 139 female college students. A median split on the MMPI-2 GM and GF scores by gender divided subjects into high and low groupings on the MMPI-2 gender variables. Multivariate analyses revealed that scores on the MMPI-2, TSCS, and SCL-90-R consistently differed by GM and GF status with differences most pronounced for the GM scale. Higher scores on the GM variable were associated with scores on the MMPI-2, TSCS, and SCL-90-R that reflected less psychopathology. Similar trends were noted for higher scorers on the GF variable, but fewer significant differences across the scales were found. The results are consistent with an interpretation of GM and GF as correlates of psychological wellbeing. PMID- 9403392 TI - Secondary trauma: assessing inter-generational transmission of war experiences with a modified Stroop procedure. AB - Intergenerational transmission of war experiences was assessed using a modified Stroop task. Adult children of war veterans and those of nonveterans named the colors in which war related words were printed. They also named the colors of neutral, positive, and OCD related words in addition to color naming a series of zeros contained on a control card. All participants completed the MMPI-II PTSD Scale, the Impact of Event Scale, and a demographic questionnaire. A statistically significant difference between the children of veterans and nonveterans was found only on the Stroop card containing war related words. Results suggest that the modified Stroop task is a sensitive measure that may have value in assessing transmission of war experiences from parents to children. PMID- 9403393 TI - A moratorium on research using the Jenkins Activity Survey for Type A behavior? AB - This article presents a brief review of literature which indicates that it may be premature to abandon the use of the Jenkins Activity Survey (JAS) for assessing Type A behavior. In particular, the literature suggests that the time urgency/irritability subcomponent of the JAS may represent a nonspecific susceptibility factor for general ill health. Moreover, studies demonstrate that the time urgency/irritability facet of the JAS is associated with a negative health-risk profile which consists of the presence of vulnerability factors and the absence of protective factors. Finally, results of this brief review are consistent with previous research with paper and pencil measures of Type A behavior showing that subcomponent measures are superior to global measures. PMID- 9403394 TI - Perceptions of the media in a community exposed to serial murder. AB - This study examined resident perceptions of the media in a community exposed to serial murder. Residents were surveyed regarding the popularity, accuracy and trust of various information sources. One hundred eighty-four residents (48% response rate) responded to mailed questionnaires. Results indicated that traditional sources of news (television, radio, and newspapers) were the most popular sources of information. However, police press conferences were judged to be the most accurate and trustworthy. Residents viewed media reports on methods of reducing personal risk as beneficial and enhancing feelings of safety. However, details of mutilations and sensational reporting were judged to increase personal fears and led to widespread dissatisfaction with the media. PMID- 9403395 TI - The efficacies of three relaxation regimens in the treatment of PTSD in Vietnam War veterans. AB - Ninety male Vietnam veterans with posttraumatic stress disorder (PTSD) were administered relaxation instructions, relaxation instruction with deep breathing exercises, or relaxation instructions with deep breathing training and thermal biofeedback. Improvement appeared on only 4 of the 21 PTSD and physiological dependent variables studied. All 21 Treatment x Time interactions were nonsignificant. This suggests that the treatments were mildly therapeutic, but that the additions of training in deep breathing and thermal biofeedback did not produce improvement beyond that associated with simple instructions to relax in a comfortable chair. PMID- 9403396 TI - Psychotherapeutic change is predictable, spontaneous change is not. AB - As a test of the possibility clinically to predict psychotherapy outcome for individual patients, eight psychotherapist judges rated 30 patients for suitability for psychotherapy and predicted individual outcomes of psychotherapy. Unknown to the judges, 15 of the patients had been in psychotherapy (T group), whereas the remaining 15 had not (NT group), but all patients had been rated with respect to change. The hypothesis was confirmed that judges would be able to predict change in the T group better than in the NT group, but that their suitability ratings would be equally predictive of change in the two groups. It was concluded that psychotherapeutic change was a systematic effect over and above initial status, large with some patients, small with others, but reliably so. PMID- 9403397 TI - The relationship of male self-report of rape supportive attitudes, sexual fantasy, social desirability and physiological arousal to sexually coercive stimuli. AB - Studies have supported the finding that sexually coercive behavior exists between males and females on college campuses and that when women say "no" to sexual behavior, men do not believe them. This study utilized penile plethysmography to investigate male sexual arousal to rape myth scenarios in a college population. The scenarios portrayed a woman who said "no" to sexually coercive intercourse behavior by a male. Results indicated that a low level of social desirability, sexual fantasies involving group sexual activity, as well as hurting and being hurt by a partner were associated with greater levels of physiological responding to coercive stimuli. Supportive attitudes about rape showed no relationship with physiological responding, yet did correlate with the sexual fantasy of being hurt by a partner, which was itself related to increased sexual arousal to sexually coercive audio stimuli. PMID- 9403398 TI - Assessing PTSD with the Millon Clinical Multiaxial Inventory-III. AB - We studied the utility of the Millon Clinical Multiaxial Inventory-III (MCMI-III) in assessing substance-abusing (n = 228), combat-related PTSD patients (n = 32). The MCMI-III produced a code type (16A) that was quite different from MCMI-I and MCMI-II code types (8A2) among similar populations. The PTSD Scale (R) successfully differentiated between a PTSD and non-PTSD substance-abusing group using mean Base Rate scores, was the best predictor of PTSD in a multiple regression equation, and the scale's sensitivity and specificity in detecting and/or ruling out the disorder was above that provided by chance alone and higher than the values reported in the test manual for that scale. The MCMI-III may be used as a broad band screening instrument for PTSD, at least among patients with combat-related stress. PMID- 9403399 TI - Do physicians discuss HIV and AIDS with patients? A survey of physician practices. AB - HIV and AIDS continue to be major concerns to the health care community and the world around them. Preventive efforts and education have been the focus of the fight against AIDS thus far. By the year 2000, 75% of physicians are expected to conduct risk-reduction counseling for patients regularly. Previous studies show that a smaller percentage "routinely" follow this recommendation. The purpose of our study was to assess with what percentage of patients physicians discuss several HIV/ AIDS-related topics, what percentage of their patients are considered at risk for infection, and how comfortable the physicians are with their knowledge level and discussing the subject matter. We sent surveys to the last five graduating classes from St. Louis University School of Medicine and to 169 physician preceptors in the community. The survey asked about patients considered at risk, physician comfort level with HIV/ AIDS, the percentage of patients they discuss various HIV/AIDS topics with, and his or her preparedness for these discussions. Total responses were 464 (53.7%) representing all areas of medicine. Most of the physicians (72.9%) consider 0-25% of their patients at risk for HIV/AIDS. Eighty-one percent claim they are moderately or very comfortable discussing the material with patients and more than 90% feel they have at least adequate knowledge. Most of the respondents discuss the HIV/ AIDS topics with 0 25% of patients. Recent medical school graduates and primary care physicians are more comfortable with HIV/AIDS and discuss the surveyed topics with a higher percentage of patients. PMID- 9403400 TI - Preventive services in a Medicare managed care environment. AB - The results of a four year demonstration project of preventive services for Medicare managed care enrollees suggest that health promotion programs can impact health behaviors and outcomes. The study provided selected preventive services to 1,800 Medicare enrollees in a managed care environment. Participants were randomly assigned to control and experimental groups with the experimental group receiving an intervention service package and the control group usual care. The results included enhanced health behavior practices, lower depression, and higher immunization rates among those individuals in the experimental group. This study suggests that selected preventive services can be provided in a managed care environment to Medicare enrollees with likely positive health status and utilization outcomes. PMID- 9403401 TI - Heart disease education and prevention program targeting immigrant Latinos: using focus group responses to develop effective interventions. AB - Although research has provided considerable knowledge concerning the positive effects of behavioral change on morbidity and mortality from heart disease and related risk factors, some segments of the population have not benefited equitably from this information. In April 1995, the National Heart, Lung, and Blood Institute (NHLBI) conducted seven focus groups to determine knowledge and attitudes about heart disease and associated risk factors, identify media usage and preferences, and assess publications usage and preferences among Spanish speaking Latino immigrants residing in the Washington, D.C., metropolitan area. This information was gathered to assist in the development of key messages and strategies for the NHLBI Latino Community Cardiovascular Disease Prevention and Outreach Initiative, Salud para su Corazon--a heart disease prevention and education campaign. Findings from these focus groups indicate that Latinos may not benefit from heart disease prevention messages developed for the general population because of language and cultural differences. The researchers concluded that health education and disease prevention programs targeting the Latino community should develop educational materials and interventions that address language preferences and cultural values. Furthermore, to be effective, these programs should show people how to make positive behavioral changes based on their current circumstances, while remaining sensitive to the fact that Latino immigrants face major life adjustments and many are still greatly influenced by their country of origin. PMID- 9403402 TI - The "Yes, I Quit" smoking cessation course: does it help women in a low income community quit? AB - The objectives were to evaluate the impact of "Yes, I Quit" (a smoking cessation course tailored for women in a low income, low education community), and to identify baseline predictors of short and longer-term self-reported cessation. The impact was evaluated in a before-after study design with no comparison group. Baseline data were collected in self-administered questionnaires at the beginning of the first session of the course. Follow-up data were collected in telephone interviews at one, three and six months after the designated Quit Day. Self reported quit rates among 122 participants were 31.1%, 24.7% and 22.3% at one, three and six months. Non-quitters reduced their consumption by 10.3, 8.3, and 7.1 cigarettes per day at one, three and six months. Multivariate logistic regression analyses showed that being in excellent/good health was significantly associated with cessation at one month (odds ratio (OR) = 2.4). Being married (OR = 13.0) and no other smokers in the household (OR = 3.6) were associated with three-month cessation. Only being married was associated with six-month cessation (OR = 6.8). "Yes, I Quit" produced quit rates among low income, low education participants comparable to those reported for cessation programs directed at the general population of smokers. Good health is associated with early cessation, while support from a spouse is important to maintaining a non-smoking status among quitters. PMID- 9403403 TI - Pharmacists' opinions and practices related to the sale of cigarettes and alcohol -a follow-up study. AB - Opinions that pharmacists hold and their practices concerning sale of cigarettes and alcohol are of interest to health experts. As a follow-up to a 1990 statewide survey of pharmacists opinions and practices related to the sale of cigarettes and alcohol, this study was designed (1) to determine current opinions and practices of pharmacists' related to the sale of cigarettes and alcohol; (2) compare these findings with results from the 1990 study; and (3) to gather new information on pharmacists' practice of health promotion activities. A structured survey questionnaire was designed and reviewed by a jury of experts and subsequently administered to half of the 1340 pharmacies in Indiana. Collected data were analyzed by using descriptive and inferential statistical methods. Findings reveal that 64 percent of responding pharmacists sell cigarettes in their stores even though 82 percent think that their stores should not sell cigarettes. Approximately 42 percent of the pharmacies sell alcoholic beverages while more than two-thirds of the pharmacists (68%) think pharmacies should not sell alcoholic beverages. These findings represent a decline of 7.2 percent in pharmacies that sell cigarettes and a 6.8 percent increase in pharmacies selling alcoholic beverages compared to the 1990 study. Study results also revealed that most pharmacists agree the use of cigarettes and alcohol were important causes of morbidity and mortality and that pharmacists should play a role in health promotion and disease prevention to the public. However, the majority do not ask their patients about their smoking and alcohol habits and do not participate in health education/promotion programs for the general community. PMID- 9403404 TI - A clinimetric approach to the components of the patient-physician relationship. AB - Although patient-physician relationships have been expressed with diverse concepts and models, we have formulated a clinimetric classification derived from several years of observation and discussions at weekly house-staff conferences devoted to "difficult" patients. The observed phenomena are classified into the following components: (1) background factors intrinsic to patient and physician before they meet, (2) individual anticipations and hopes for what may happen, (3) extrinsic features of the setting, (4) individual reactions during the encounter, and (5) the consequences thereafter. These interacting components are usually too complex for characterizations based on single models for the relationship or single titles (such as "hateful" or "noncompliant") for the patient. The components can serve as a "review of systems" for identifying manifestations, sources, and solutions to such common problems as discordant hopes, the physician's unawareness of the patient's pertinent extramedical status, psychiatric and mental-status challenges, and cogent factors in chronic illness. PMID- 9403405 TI - Worry grows as antibiotic-resistant bacteria continue to gain ground. PMID- 9403406 TI - Studies suggest a darker side of 'benign' microbes. PMID- 9403407 TI - From the Centers for Disease Control and Prevention. Impact of promotion of the Great American Smokeout and availability of over-the-counter nicotine medications, 1996. PMID- 9403408 TI - From the Centers for Disease Control and Prevention. State-specific prevalence of cigarette smoking among adults, and children's and adolescents' exposure to environmental tobacco smoke--United States, 1996. PMID- 9403410 TI - A piece of my mind. Whispering through the keyhole. PMID- 9403409 TI - From the Centers for Disease Control and Prevention. Medical-care expenditures attributable to cigarette smoking during pregnancy--United States, 1995. PMID- 9403411 TI - Mortality and length of stay in teaching vs nonteaching hospitals. PMID- 9403412 TI - Mortality and length of stay in teaching vs nonteaching hospitals. PMID- 9403413 TI - Mortality and length of stay in teaching vs nonteaching hospitals. PMID- 9403414 TI - Prepublication of NIH Consensus Conferences on the Internet. PMID- 9403415 TI - Correction and clarification: factors contributing to the death of General Stonewall Jackson. PMID- 9403416 TI - Early discharge of newborns. PMID- 9403417 TI - Early discharge of newborns. PMID- 9403418 TI - Early discharge of newborns. PMID- 9403419 TI - Early discharge of newborns. PMID- 9403420 TI - Risk of end-stage renal disease in diabetes mellitus: a prospective cohort study of men screened for MRFIT. Multiple Risk Factor Intervention Trial. AB - CONTEXT: Diabetes is a frequent cause of end-stage renal disease (ESRD). However, the degree of risk is uncertain. OBJECTIVE: To determine the relative risk (RR) of ESRD related to diabetes in the United States. DESIGN: Nonconcurrent prospective cohort study. PARTICIPANTS: A total of 332544 men aged 35 to 57 years from 18 US cities screened in 1973 to 1975 for participation in the Multiple Risk Factor Intervention Trial (MRFIT). MAIN EXPOSURE: Diabetes mellitus defined by self-reported use of medication for diabetes at baseline. MAIN OUTCOME: Incident ESRD through 1990 assessed from a national ESRD registry and by surveillance for death from renal disease. RESULTS: Over an average follow-up of 16 years, there were 136 cases of ESRD in 5147 diabetic men and 678 cases in 327397 nondiabetic men. Age-adjusted incidence of all-cause ESRD in the diabetic men was 199.8 per 100000 person-years compared with 13.7 per 100000 person years in their nondiabetic counterparts (RR, 12.7; 95% confidence interval [CI], 10.5-15.4). Diabetic men were also at higher risk for ESRD ascribed to causes other than diabetes (RR=4.3; 95% CI, 3.2-5.9). With simultaneous adjustment for age, ethnicity, income, blood pressure, serum cholesterol level, and history of myocardial infarction, diabetic men remained at higher risk for all-cause ESRD (RR, 9.0; 95% CI, 7.4-11.0), ESRD ascribed to diabetes (RR, 92.3; 95% CI, 64.6 131.9), and ESRD ascribed to nondiabetic causes (RR, 3.0; 95% CI, 2.2-4.1). CONCLUSIONS: Diabetes mellitus is a strong independent risk factor for ESRD, even for ESRD ascribed to causes other than diabetes. Improvements in the prevention and control of diabetes should produce substantial reductions in ESRD incidence. PMID- 9403421 TI - Implementation of the Ottawa Knee Rule for the use of radiography in acute knee injuries. AB - CONTEXT: The Ottawa Knee Rule is a previously validated clinical decision rule that was developed to allow physicians to be more selective and efficient in their use of plain radiography for patients with acute knee injuries. OBJECTIVE: To assess the impact on clinical practice of implementing the Ottawa Knee Rule. DESIGN: Controlled clinical trial with before-after and concurrent controls. SETTING: Emergency departments of 2 teaching and 2 community hospitals. PATIENTS: All 3907 consecutive eligible adults seen with acute knee injuries during two 12 month periods before and after the intervention. INTERVENTION: During the after period in the 2 intervention hospitals, the Ottawa Knee Rule was taught to all house staff and attending physicians who were encouraged to order knee radiography according to the rule. MAIN OUTCOME MEASURES: Referral for knee radiography, accuracy and reliability of the rule, mean time in emergency department, and mean charges. RESULTS: There was a relative reduction of 26.4% in the proportion of patients referred for knee radiography in the intervention group (77.6% vs 57.1 %; P<.001), but a relative reduction of only 1.3% in the control group (76.9% vs 75.9%; P=.60). These changes over time were significant when the intervention and control groups were compared (P<.001). The rule was found to have a sensitivity of 1.0 (95% confidence interval [CI], 0.94-1.0) for detecting 58 knee fractures. The K coefficient for interpretation of the rule was 0.91 (95% CI, 0.82-1.0). Compared with nonfracture patients who underwent radiography during the after-intervention period, those discharged without radiography spent less time in the emergency department (85.7 minutes vs 118.8 minutes) and incurred lower estimated total medical charges for physician visits and radiography (US $80 vs US $183). CONCLUSIONS: Implementation of the Ottawa Knee Rule led to a decrease in use of knee radiography without patient dissatisfaction or missed fractures and was associated with reduced waiting times and costs. Widespread use of the rule could lead to important health care savings without jeopardizing patient care. PMID- 9403422 TI - Quality of care, process, and outcomes in elderly patients with pneumonia. AB - CONTEXT: Pneumonia is a frequent cause of hospitalization and death among elderly patients, but the relationships between processes of care for pneumonia and outcomes are uncertain, making quality improvement a challenge. OBJECTIVES: To assess quality of care for Medicare patients hospitalized with pneumonia and to determine whether process of care performance is associated with lower 30-day mortality. DESIGN: Multicenter retrospective cohort study with medical record review. SETTING: A total of 3555 acute care hospitals throughout the United States. PATIENTS: A total of 14069 patients at least 65 years old hospitalized with pneumonia. MAIN OUTCOME MEASURES: Four processes of care: time from hospital arrival to initial antibiotic administration; blood culture collection before initial hospital antibiotics; blood culture collection within 24 hours of hospital arrival; and oxygenation assessment within 24 hours of hospital arrival. Associations between processes of care and 30-day mortality were determined with logistic regression analysis. RESULTS: National estimates of process-of-care performance were antibiotic administration within 8 hours of hospital arrival, 75.5% (95% confidence interval [CI], 73.1-77.9); blood cultures before antibiotics, 57.3% (95% CI, 54.5-60.1); initial blood culture collection, 68.7% (95% CI, 66.2-71.2); and initial oxygenation assessment, 89.3% (95% CI, 87.5 90.9). Lower 30-day mortality was associated with antibiotic administration within 8 hours of hospital arrival (odds ratio [OR], 0.85; 95% CI, 0.75-0.96) and blood culture collection within 24 hours of arrival (OR, 0.90; 95% CI, 0.81 1.00). State and territory performance estimates varied from 49.0% to 89.7% for antibiotics given within 8 hours and from 45.6% to 82.6% for blood cultures drawn within 24 hours. CONCLUSIONS: Administering antibiotics within 8 hours of hospital arrival and collecting blood cultures within 24 hours were associated with improved survival. The fact that states varied widely in the performance of these measures suggests that opportunities exist to improve hospital care of elderly patients with pneumonia. PMID- 9403423 TI - Risk factors and clinical presentation of acute primary HIV infection in India. AB - CONTEXT: Most previous studies of clinical presentation and risk factors in early human immunodeficiency virus (HIV) infection have relied on retrospective analyses and referred seroconverters, and thus were subject to possible bias. OBJECTIVES: To apply a method based on measurement of prevalent HIV-1 p24 antigenemia for identification of risk factors for newly acquired HIV infection and to describe the signs and symptoms of acute HIV infection. DESIGN AND SETTING: Nested case-control study in Pune, India. PARTICIPANTS: HIV antibody negative persons attending 2 sexually transmitted disease (STD) clinics between May 1993 and June 1996. OUTCOME MEASURES: Prevalent p24 antigenemia, risk factors for HIV infection, and clinical symptoms of acute primary HIV infection. RESULTS: Of 3874 HIV antibody-negative persons tested, 58 (1.5%) were p24 antigen positive at initial presentation to the clinics. Unprotected sexual contact with a commercial sex worker (CSW) was reported by 39 (77%) of the 51 p24 antigenemic men, compared with 131 (51 %) of 255 control men (adjusted odds ratio [AOR], 3.4; 95% confidence interval [CI], 1.2-9.6; P=.02). The presence of an active genital ulcer at the time of screening was found in 46 (79%) of the 58 p24 antigenemic men and women, compared with 137 (47%) of the 290 control subjects (AOR, 4.2; 95% CI, 2.0-9.0; P<.001). Signs and symptoms independently associated with p24 antigenemia in HIV antibody-seronegative persons included fever, which was reported by 28 (48%) of the 58 p24 antigenemic subjects, but only 52 (18%) of the 290 control subjects (AOR, 4.7; 95% CI, 2.4-9.0; P<.001). Joint pain was reported by 10% of subjects recently HIV infected, compared with 2% of the control subjects (AOR, 6.5; 95% CI, 1.7-24.8; P=.006). Night sweats were reported by 9% of the p24 antigenemic, but only 1% of the control subjects (AOR, 9.1; 95% CI, 1.7-47.6; P=.009). Overall, fever, joint pain, and/or night sweats were reported in 27 (47%) of the 58 subjects with recent HIV infection. CONCLUSIONS: This systematic case-control study of p24 antigen screening in HIV-seronegative patients attending STD clinics in India identified unprotected sex with a CSW and a genital ulcer as independent risk factors associated with newly acquired HIV infection. In addition, p24 antigen positivity identified recent fever, night sweats, and arthralgias as symptoms that may be predictive of recent HIV infection. In a study of patients attending STD clinics in India, screening for p24 antigen in HIV antibody-negative persons was found to be a reliable and effective research method for determining recent risk behavior and identifying clinical signs of acute primary HIV infection. PMID- 9403424 TI - Spread of HIV infection in married monogamous women in India. AB - CONTEXT: A high prevalence of human immunodeficiency virus (HIV) infection in female sex workers (FSWs) and men who attend sexually transmitted disease (STD) clinics poses a risk for spread of infection to other populations. OBJECTIVE: To examine spread of HIV to a low-risk population by comparing prevalence of, and risk factors for, HIV and STDs in FSWs and non-FSWs. METHODS: Women attending STD clinics in Pune, India, were assessed for STDs and HIV from May 13, 1993, to July 11, 1996. Demographic and behavioral information was collected, and clinical and laboratory assessment was performed. MAIN OUTCOME MEASURE: Prevalence and risk determinants of HIV infection. RESULTS: Of 916 women enrolled, 525 were FSWs and 391 were non-FSWs. Prevalence of HIV in FSWs and non-FSWs was 49.9% and 13.6%, respectively (P<.001). In multivariate analysis, inconsistent condom use and genital ulcer disease or genital warts were associated with prevalent HIV in FSWs. History of sexual contact with a partner with an STD was associated with HIV in non-FSWs. CONCLUSIONS: Infection with HIV is increasing in non-FSWs, previously thought to be at low risk in India. Since history of sexual contact with their only sex partner was the only risk factor significantly associated with HIV infection, it is likely that these women are being infected by their spouses. This underscores the need for strengthening partner-notification strategies and counseling facilities in India. PMID- 9403425 TI - Comparison of primary coronary angioplasty and intravenous thrombolytic therapy for acute myocardial infarction: a quantitative review. AB - OBJECTIVE: To provide a quantitative review of the treatment effects of primary coronary angioplasty vs intravenous thrombolysis for acute myocardial infarction. DATA SOURCES: Ten randomized trials were identified through computerized bibliographic search of MEDLINE from January 1985 through March 1996 and by queries of principal investigators. STUDY SELECTION: Single-center and multicenter randomized trials comparing primary angioplasty with intravenous thrombolytic therapy among 2606 patients were included. Four trials compared angioplasty with streptokinase, 3 compared angioplasty with a 3- to 4-hour infusion of tissue-type plasminogen activator, and 3 compared angioplasty with "accelerated" administration of tissue-type plasminogen activator over 90 minutes. DATA EXTRACTION: Each investigator provided definitions and exact data for outcome events. Odds ratios (ORs), 95% confidence intervals (CIs), and P values were calculated using exact tests for categorical data. DATA SYNTHESIS: Mortality at 30 days or less was 4.4% for the 1290 patients treated with primary angioplasty compared with 6.5% for the 1316 patients treated with thrombolysis (34% reduction; OR, 0.66; 95% CI, 0.46-0.94; P=.02). The effect was similar among thrombolytic regimens, and no subgroup demonstrated a significant reduction in death. The rates of death or nonfatal reinfarction were 7.2% for angioplasty and 11.9% for thrombolytic therapy (OR, 0.58; 95% CI, 0.44-0.76; P<.001). Angioplasty was associated with a significant reduction in total stroke (0.7% vs 2.0%; P=.007) and hemorrhagic stroke (0.1% vs 1.1%; P<.001). CONCLUSIONS: Based on outcomes at hospital discharge or 30 days, primary angioplasty appears to be superior to thrombolytic therapy for treatment of patients with acute myocardial infarction, with the proviso that success rates for angioplasty are as good as those achieved in these trials. Data evaluating longer-term outcomes, operator experience, and time delay before treatment are needed before primary angioplasty can be universally recommended as the preferred treatment. PMID- 9403426 TI - Palliative options of last resort: a comparison of voluntarily stopping eating and drinking, terminal sedation, physician-assisted suicide, and voluntary active euthanasia. AB - Palliative care is generally agreed to be the standard of care for the dying, but there remain some patients for whom intolerable suffering persists. In the face of ethical and legal controversy about the acceptability of physician-assisted suicide and voluntary active euthanasia, voluntarily stopping eating and drinking and terminal sedation have been proposed as ethically superior responses of last resort that do not require changes in professional standards or the law. The clinical and ethical differences and similarities between these 4 practices are critically compared in light of the doctrine of double effect, the active/passive distinction, patient voluntariness, proportionality between risks and benefits, and the physician's potential conflict of duties. Terminal sedation and voluntarily stopping eating and drinking would allow clinicians to remain responsive to a wide range of patient suffering, but they are ethically and clinically more complex and closer to physician-assisted suicide and voluntary active euthanasia than is ordinarily acknowledged. Safeguards are presented for any medical action that may hasten death, including determining that palliative care is ineffective, obtaining informed consent, ensuring diagnostic and prognostic clarity, obtaining an independent second opinion, and implementing reporting and monitoring processes. Explicit public policy about which of these practices are permissible would reassure the many patients who fear a bad death in their future and allow for a predictable response for the few whose suffering becomes intolerable in spite of optimal palliative care. PMID- 9403427 TI - Preserving scientific debate and patient choice: lessons from the Consensus Panel on Mammography Screening. National Institutes of Health. PMID- 9403428 TI - Translating good advice into better practice. PMID- 9403429 TI - Primary angioplasty compared with thrombolytic therapy for acute myocardial infarction. PMID- 9403430 TI - Effect of CCR2 and CCR5 variants on HIV disease: abstract and commentary. PMID- 9403431 TI - Derek F. C. Harwood Nash, MB, ChB, DSc, FRCPC, FACR, FC RAD (SA). PMID- 9403432 TI - Can academic neuroradiology survive in this era of managed care? PMID- 9403433 TI - The encephalopathic neonate: choosing the proper imaging technique. PMID- 9403434 TI - The roles of MR angiography, CT angiography, and sonography in vascular imaging of the head and neck. PMID- 9403435 TI - The treatment of acutely ruptured cerebral aneurysms: endovascular therapy versus surgery. PMID- 9403436 TI - Prethrombolysis brain imaging: trends and controversies. PMID- 9403437 TI - Rule out eighth nerve tumor: contrast-enhanced T1-weighted or high-resolution T2 weighted MR? PMID- 9403438 TI - Cryptic vascular malformations: controversies in terminology, diagnosis, pathophysiology, and treatment. PMID- 9403439 TI - Imaging the brain in dementia: expensive and futile? PMID- 9403440 TI - Screening sinus CT: a good idea gone bad? PMID- 9403441 TI - Do the benefits of image guidance in neurosurgery justify the costs? From stereotaxy to intraoperative MR. PMID- 9403442 TI - The proper terminology for reporting lumbar intervertebral disk disorders. PMID- 9403443 TI - Controversies in imaging acute cervical spine trauma. PMID- 9403444 TI - The present controversy over the imaging method of choice for evaluating the soft tissues of the neck. PMID- 9403445 TI - The value of proton MR spectroscopy in pediatric metabolic brain disease. PMID- 9403446 TI - Cardiac rupture complicating cerebral intraarterial thrombolytic therapy. AB - We report a case of fatal cardiac rupture occurring during intraarterial thrombolytic therapy for acute embolic stroke in a patient with recent myocardial infarction. PMID- 9403447 TI - Failure as teacher. PMID- 9403448 TI - Foreign bodies in small arteries after use of an infusion microcatheter. AB - Over a 31-month period, we performed four neurointerventional procedures after which unexpected foreign bodies were noted in multiple arteries. All four procedures had in common the use of Fastracker-18 infusion microcatheters. Histologically, the intravascular debris looked strikingly similar to the hydrophilic coating on the catheter. An in vitro test mimicking clinical use of the microcatheter revealed that the hydrophilic coating can separate from the catheter. Until the coating is refined to make it more resistant to stripping, it may be advisable to reduce the amount of back-and-forth movement of these microcatheters if they have been positioned through guide catheters with small inner diameters and angled tips. PMID- 9403449 TI - The use of hydrophilic catheters in small arteries. PMID- 9403450 TI - An arteriovenous malformation model for testing liquid embolic materials. AB - An arteriovenous malformation model with a transparent nidus was constructed to investigate the embolization behavior of polyvinyl acetate (PVAc) solution relative to flow velocity in the feeding artery and injection speed. It was found that the liquid embolic material flowed distally when injected too fast or when the flow velocity was too low. In addition, we found that solution consisting of PVAc plus metrizamide was a better embolic material than solution containing only PVAc. PMID- 9403451 TI - Percutaneous polymethylmethacrylate vertebroplasty in the treatment of osteoporotic vertebral body compression fractures: technical aspects. AB - PURPOSE: To describe a technique for percutaneous vertebroplasty of osteoporotic vertebral body compression fractures and to report early results of its use. METHODS: The technique was used over a 3-year period in 29 patients with 47 painful vertebral fractures. The technique involves percutaneous puncture of the involved vertebra(e) via a transpedicular approach followed by injection of polymethylmethacrylate (PMMA) into the vertebral body. RESULTS: The procedure was technically successful in all patients, with an average injection amount of 7.1 mL PMMA per vertebral body. Two patients sustained single, nondisplaced rib fractures during the procedure; otherwise, no clinically significant complications were noted. Twenty-six patients (90%) reported significant pain relief immediately after treatment. CONCLUSION: Vertebroplasty is a valuable tool in the treatment of painful osteoporotic vertebral fractures, providing acute pain relief and early mobilization in appropriate patients. PMID- 9403452 TI - Paradoxical hyperfixation of hexamethylpropyleneamine oxime in cerebral infarction. AB - We describe four patients with cerebral infarction and 99mTc hexamethylpropyleneamine oxime (HMPAO) hyperfixation in distributions corresponding to the infarctions seen at CT and/or MR imaging. Increased HMPAO extraction due to hyperpermeability across the blood-brain barrier and increased retention due to reduced back diffusion of the tracer probably accounted for the increased fractional fixation in infarcts seen in our patients. PMID- 9403453 TI - Proton MR spectroscopy and neuropsychological testing in adrenoleukodystrophy. AB - PURPOSE: To determine early signs of disease in patients with childhood-onset cerebral adrenoleukodystrophy (COCALD) with the use of proton MR spectroscopy. METHODS: Eleven children with posterior COCALD involvement and three children with anterior COCALD involvement were studied with single-voxel proton MR spectroscopy and neuropsychological testing. Findings were compared with those in five healthy control subjects. RESULTS: Areas of abnormal T2 signal intensity in children with COCALD showed abnormal metabolite ratios relative to those of control subjects as follows: decreased N-acetylaspartate (NAA)/Creatine (Cr) and NAA/Choline (Ch) and increased Ch/Cr. Metabolite ratios from normal-appearing brain regions in the same patients also were abnormal, with reduced NAA/Cr and NAA/Ch and increased Ch/Cr values. The mean metabolite ratios in normal-appearing regions were between those in the abnormal regions and those found in the control subjects. Statistical comparison of these ratios with neuropsychological test scores, which are specific for anterior and posterior brain functions, showed a significant correlation with the abnormal metabolite ratios. Our results indicate that the normal-appearing brain regions in these patients are metabolically abnormal. CONCLUSION: Proton MR spectroscopy could be a useful noninvasive tool to evaluate extent of disease in patients with COCALD. PMID- 9403454 TI - Congenital ocular motor apraxia: imaging findings. AB - PURPOSE: To determine the frequency of cerebellar and cerebral abnormalities on brain imaging studies in children with congenital ocular motor apraxia. METHODS: Brain imaging studies were performed in 19 children with typical congenital ocular motor apraxia who were in the care of a visual impairment program at a children's hospital. Independent clinical review categorized the subjects as having partial (n = 10) or expanded (n = 9) congenital ocular motor apraxia on the basis of extent of associated speech or neurodevelopmental problems. Fifteen CT studies and 13 MR examinations of the brain performed in these children were reviewed independently by two pediatric neuroradiologists. Radiologic findings were agreed on by consensus. RESULTS: Cerebellar abnormalities were found in 12 of 19 cases. The cerebellar vermis was small in 10 children. A small cerebellar vermis was the only abnormality in five of 10 children with partial congenital ocular motor apraxia and in two of nine children with expanded congenital ocular motor apraxia. Among seven children with a small vermis examined with high resolution MR imaging, the inferior portion of the vermis was preferentially involved in each case. Of these seven subjects, none of four with partial congenital ocular motor apraxia but two of three with expanded congenital ocular motor apraxia had an abnormality of the superior portion of the vermis. Miscellaneous supratentorial lesions affecting both gray and white matter were found in six subjects. Five of the 19 children had normal imaging findings. CONCLUSION: Inferior vermian hypoplasia is the most common abnormality in children with congenital ocular motor apraxia. PMID- 9403455 TI - GRASE (gradient- and spin-echo) MR of the brain. AB - PURPOSE: To assess the clinical utility of GRASE (gradient- and spin-echo) MR imaging of the brain by comparing it with the T2-weighted turbo spin-echo technique. METHODS: Fifty-three consecutive patients referred for MR imaging of the brain were studied with T2-weighted turbo spin-echo and GRASE techniques, matched for effective echo time (110 milliseconds), echo train length (eight), and spatial resolution. The examinations were evaluated independently by two neuroradiologists for lesion detection (high- and low-signal-intensity lesions) and lesion conspicuity, and for susceptibility, motion, and chemical-shift artifacts. RESULTS: The GRASE technique provided greater detection of both high- and low-signal-intensity lesions and of low-signal-intensity lesions with paramagnetic susceptibility characteristics (ie, calcium and hemorrhage). Chemical-shift artifacts in the frequency-encoding direction were more prominent with the turbo spin-echo technique, whereas chemical-shift artifacts in the phase encoding direction were more prominent with the GRASE technique. There was no significant difference in the degree of diamagnetic susceptibility artifacts at the base of the skull, or in motion artifacts. CONCLUSION: T2-weighted GRASE is a fast imaging technique with a potential for replacing turbo spin-echo in routine MR imaging of the brain. GRASE maintains the contrast resolution of turbo spin echo imaging and is better at depicting lesions with paramagnetic susceptibility characteristics. PMID- 9403456 TI - Changes in blood flow velocity and diameter of the middle cerebral artery during hyperventilation: assessment with MR and transcranial Doppler sonography. AB - PURPOSE: To compare blood flow velocity changes within the middle cerebral artery (MCA) during hyperventilation, as measured with by both transcranial Doppler sonography and MR imaging, with the diameter of the MCA as measured with MR imaging alone. METHODS: The studies were performed in six healthy volunteers ranging in age from 22 to 31 years (mean, 27 years). Transcranial Doppler sonography was carried out with a range-gated 2-MHz transducer. MR examinations were done on a 1.5-T imaging unit. MR angiography was performed using the time-of flight technique. MR flow measurements were carried out by using the phase mapping technique with an ECG-triggered phase-contrast sequence. RESULTS: During hyperventilation, the mean blood flow velocity of the proximal MCA declined by 49.6% +/- 5.7 (mean +/- standard deviation) as measured with Doppler sonography, and by 47% +/- 4.6 as measured with MR flow calculation. The diameter of the MCA (3.4 +/- 0.3 mm) remained unchanged on MR imaging studies (3.3 +/- 0.3 mm). CONCLUSION: We found a good correlation between relative flow velocity changes measured by transcranial Doppler sonography and MR techniques. MR imaging revealed no significant changes in the diameter of the proximal MCA during normal versus hyperventilation. Relative changes in flow velocity in the MCA would thereby reflect relative changes in cerebral blood flow, at least during hyperventilation. PMID- 9403457 TI - Cerebral complications of murine monoclonal CD3 antibody (OKT3): CT and MR findings. AB - Treatment of acute renal allograft rejection with mouse monoclonal antibody (OKT3) is associated with systemic and neurologic side effects. We describe cerebral abnormalities in a 13-year-old boy with steroid-resistant renal allograft rejection. After treatment with OKT3, an acute neurologic syndrome developed, including seizures, lethargy, and decreased mental function. CT and MR imaging revealed confluent cerebral lesions at the corticomedullary junction. Contrast-enhanced MR images showed patchy enhancement, indicating blood-brain barrier dysfunction. The diagnosis of OKT3-induced encephalopathy with cerebral edema and capillary leak syndrome was made. Although CT and MR findings are nonspecific, neuroradiologists should be aware of this condition in transplant patients treated with OKT3. PMID- 9403458 TI - T2-weighted three-dimensional turbo spin-echo MR of intracranial aneurysms. AB - A T2-weighted three-dimensional turbo spin-echo sequence is described that accurately depicted the anatomy of 18 intracranial aneurysms in 10 patients. PMID- 9403459 TI - True malignant mixed tumor (carcinosarcoma) of tonsillar minor salivary gland origin: diagnostic imaging and endovascular therapeutic embolization. AB - We report a case of carcinosarcoma of the minor salivary glands of the left palatine tonsil, an especially rare location. Imaging characteristics assessed at CT, MR imaging, and angiography are presented. In addition, we describe our experience with preoperative therapeutic endovascular embolization of this hypervascular tumor. PMID- 9403460 TI - Self-induced ethmoidectomy from rhinotillexomania. AB - A 53-year-old woman with a long history of compulsive nose picking (rhinotillexomania) presented with a large, self-inflicted nasal septal perforation and right-sided penetration of the ethmoidal sinus, or "ethmoidectomy." PMID- 9403461 TI - New vistas in occupational health. PMID- 9403462 TI - Occupational health and safety in national development--the case of Australia. AB - Over the last 15 years occupational health and safety has undergone a rapid transformation in Australia. This review discusses the changes, emphasizing, the sociopolitical and economic context, national developments in policy and practice, and the dialogue between the public and occupational health and safety agencies and professionals about occupational health and safety matters. First came the classical phase when activities followed the accepted hygienic, medical, and inspection traditions laid down early in the century. A phase followed in which modern legislation was introduced, new institutions were created, and research and data gathering on important issues was intensified. Finally came the current phase characterized by a performance paradigm. Emphasis was given to regulatory reform and the use of nonregulatory initiatives to facilitate better occupational health and safety. PMID- 9403463 TI - Mortality from cardiovascular diseases and sudden death in ferroalloy plants. AB - OBJECTIVES: The aim of this study was to examine mortality from circulatory diseases and sudden death among workers in 12 Norwegian ferroalloy plants. METHODS: The cohort comprised 14730 men employed for the first time during 1933 1990 and for at least 6 months. Deaths observed during 1962-1990 were compared with expected figures calculated from national mortality rates. Internal comparisons of rates were performed by Poisson regression analysis. RESULTS: The overall mortality from cardiovascular diseases was not increased [standardized mortality ratio (SMR) 1.01], but a significantly increased mortality from sudden death (SMR 1.55) and hypertensive disease (SMR 1.37) was observed. Among the ferromanganese/silicomanganese (FeMn/SiMn) furnace workers the sudden death mortality was significantly increased during the employment period (SMR 2.47). In an internal comparison of the sudden death rates, a significant increase of 0.05 in the rate ratio per workyear was observed in this group. The mortality from 3 hypertension-related diseases combined (cerebrovascular, hypertensive, and renal diseases) showed identical positive mortality trends among the ferrosilicon/silicon-metal (FeSi/Si-met) and the FeMn/SiMn furnace workers by increasing duration of work. CONCLUSIONS: Increased mortality from sudden death among the FeMn/SiMn furnace workers is not likely to be explained by smoking or alcohol consumption. Associations with work exposures (manganese and possibly carbon monoxide and heat) are suspected. The increasing mortality from hypertension-related diseases with increasing duration of work in both groups of furnace workers may be associated with common furnace work conditions (eg, heat, psychosocial stress, shift work, noise, carbon monoxide). PMID- 9403464 TI - Mortality from nonmalignant respiratory diseases among male workers in Norwegian ferroalloy plants. AB - OBJECTIVES: This study examined mortality from nonmalignant respiratory diseases among ferroalloy workers. METHODS: The cohort comprised 14730 men employed for the first time in 1933-1990 and for at least 6 months in 1 of 12 plants. The duration of work in specific departments and exposure to amorphous silica in the ferrosilicon/silicon-metal (FeSi/Si-met) plants, estimated from a job-exposure matrix, were the main exposure variables. Deaths were observed during 1962-1990. The mortality was analyzed with the use of standardized mortality ratios (SMR) and internal comparisons of rates. RESULTS: Overall mortality from nonmalignant respiratory diseases was not increased, but mortality from bronchitis, emphysema, and asthma combined was significantly increased among the men with at least 3 years of FeSi/Si-met furnace work (SMR 1.82, 16 deaths). A Poisson regression analysis of the mortality from these causes among 6359 employees in the FeSi/Si met plants showed a significant increase of 0.06 per unit of amorphous silica exposure observed 10-20 years after the exposure. Six men died of pneumonia while still employed in a ferromanganese/silicomanganese (FeMn/SiMn) plant. No corresponding deaths occurred among employees in FeSi/Si-met plants. Only 2 deaths from pneumoconiosis were observed in the total cohort. CONCLUSIONS: Among employees in FeSi/Si-met plants increased mortality from bronchitis, emphysema, and asthma may be associated with previous exposure to amorphous silica. Deaths from pneumonia among FeMn/SiMn workers may be associated with manganese exposure. PMID- 9403465 TI - Respiratory symptoms, across-shift lung function changes and lifetime exposures of welders in New Zealand. AB - OBJECTIVES: The possibility was investigated that exposure to welding fumes causes an acute decrease in pulmonary function. METHODS: Changes in the pulmonary function of 62 current welders and 75 nonwelders at the same sites and the relationship with work-related symptoms were recorded. RESULTS: Work-related cough was reported by 22.6% of the current welders and 6.7% of the nonwelders [odds ratio (OR) 4.1, 95% confidence interval (95% CI) 1.5-11.5], and the respective figures for at least 1 work-related symptom were 30.7% and 16.0% (OR 2.3, 95% CI 1.0-5.2). The groups' preshift lung function did not differ. The mean percentage change from the base-line value of the forced expiratory volume in 1 s (FEV1.0) after 15 minutes of work was -2.8% (range -29.3% to +20.9%) for the current welders and +1.0% (range -20.7% to +22.6%) for the nonwelders (P = 0.01). A multivariate analysis identified current welding as the most significant risk factor for a decrease of at least 5% in FEV1.0 after 15 minutes, the risk being greater for those with no local exhaust ventilation (OR 22.4, 95% CI 4.5-113.4) than for those with personal protection only (OR 16.0, 95% CI 2.1-122.9) and those with local exhaust ventilation (OR 2.8, 95% CI 0.2-41.2) or passive exposure only (OR 2.0, 95% CI 0.5-8.8). CONCLUSIONS: An acute decrease in FEV1.0 in relation to work is more prevalent among welders than among nonwelders, and is more common among welders without local exhaust ventilation than among those with it. PMID- 9403466 TI - Lead concentrations in human plasma, urine and whole blood. AB - OBJECTIVES: Blood-lead levels (B-Pb), and to some extent urinary lead (U-Pb), are the most employed measures of lead exposure and risk. However, the small fraction of lead present in plasma (usually below 1% of that in blood) is probably more relevant to lead exposure and toxicity. Nevertheless, the lead content of plasma lead (P-Pb) has only seldom been used, mainly due to analytical limitations, which have now been overcome. We examined P-Pb in occupationally exposed subjects, as well as its relationship with B-Pb and U-Pb. METHODS: Blood samples were obtained from 145 male workers, 110 of whom were employed in lead work. After a simple dilution of plasma, P-Pb was determined by inductively coupled plasma mass spectrometry. The detection limit was 0.04 microg/l, and the imprecision was 5%. RESULTS: The lead concentration ranges were 0.20-37 microg/l for P-Pb, 0.9-176 microg/l (density adjusted) for U-Pb, and 9-930 microg/l for B Pb. A close exponential relation was obtained between B-Pb and P-Pb. When B-Pb was plotted versus log P-Pb, a straight line (log P-Pb = 0.00225 x B-Pb - 0.58; r = 0.97) was obtained. Both the relation between U-Pb and P-Pb and that between U Pb and B-Pb showed a large scattering (r = 0.78 in both cases). The relation to B Pb appeared to be exponential, while that to P-Pb appeared to be linear. CONCLUSIONS: The low detection limit and good precision of P-Pb determinations make it possible to use P-Pb in assessments of lead exposure and risk. Furthermore, in relative terms, P-Pb is a more sensitive measure than B-Pb, especially at high lead levels. This development is of importance for studies of exposure, possibly also for studies of risks. PMID- 9403467 TI - Occupational and personal risk factors for carpal tunnel syndrome in industrial workers. AB - OBJECTIVES: The purpose of the study was to evaluate both nonoccupational and occupational factors associated with carpal tunnel syndrome (CTS) in industrial workers. METHODS: Sixty-five workers with CTS were compared with 65 referents matched for gender, age, and plant. The medical history and household activities of the workers and the ergonomic and organizational characteristics of the job were analyzed. RESULTS: Exertion of force over 1 kg was associated with CTS [odds ratio (OR) 9.0]. Two risk factors were related to motion repetitiveness: length of the shortest elementary operation of < or = 10 s (OR 8.8) and lack of change in tasks or lack of breaks for at least 15% of the daily worktime (OR 6.0). No posture of the upper limb was associated with CTS. Workstation design involving the manual supply of the workers (OR 5.0) and the lack of job rotation (OR 6.3) were associated with CTS. The only personal factor associated with CTS was a parity of at least 3 (OR 3.2). There was a continuous increase in the odds ratio against the number of risk factors accumulated by the workers; the odds ratio thus ranged from 5.6 when 3 of the 6 risk factors were present to > or = 90 when 4, 5, or 6 risk factors were accumulated. CONCLUSIONS: The results were in agreement with a model for CTS which included 1 personal and 5 occupational risk factors. The number of risk factors cumulated by the workers seems to be a major determinant of CTS. PMID- 9403468 TI - Possible bias from rating behavior when subjects rate both exposure and outcome. AB - OBJECTIVES: In many epidemiologic studies the subjects rate both the exposure and the outcome, assigning numerical values to the variables according to their perceptions and judgments. Hypothetically, subjects who tend to overestimate, exaggerate, or use high numerical values in rating tasks would rate both exposure and outcome higher than subjects who tend to underestimate, dissimulate, or use low numerical values. A range of such rating behaviors among the subjects would introduce uncontrollable bias to relative risk estimates, in most cases an overestimation. The aim of this study was to assess the possible presence of and effects on relative risk estimates of such high and low rating behavior among subjects in an epidemiologic study of musculoskeletal disorders. METHODS: Rating behavior was analyzed by intercorrelating the ratings of 19 different stimuli. High positive correlations would indicate the presence of high and low rating behavior. RESULTS: The correlations were, however, both positive and negative and close to zero. Adjusting for rating behavior did not affect relative risk estimates, based on subjective ratings of both exposure and outcome. CONCLUSION: There is no support in this study for the existence of a range of high and low rating behavior among subjects who rate neutral and nonaffective stimuli, such as time, weight, number and physical exposure, as well as pain and other symptoms. There is therefore no support for the idea of a bias to relative risk estimates from such rating behavior in studies where subjects rate both exposure and outcome variables of this kind. PMID- 9403469 TI - Job adjustment as a means to reduce sickness absence during pregnancy. AB - OBJECTIVES: This study examined the effect of job adjustment on sickness absence during pregnancy and also determined the conditions under which such adjustments are obtained. METHODS: Data were derived from a nationally representative survey on work conditions during pregnancy in Norway in 1989. For employees (N = 2713) remaining in the same job throughout pregnancy, the percentage of women on sick leave immediately before delivery was determined according to the need for job adjustment and the obtainment of job adjustment. Those obtaining job adjustment were grouped according to workplace size, labor-market sector, co-worker gender, educational level, work schedules, weekly workhours, children under 16 years of age in the household, and age. RESULTS: All told, 1691 women (62.3%) needed job adjustment, among whom 936 (55.4%) obtained such adjustment. The proportions of those on sick leave before delivery were 45.2% for "no need", 67.9% for "need - adjustment obtained", and 79.2% for "need - adjustment not obtained". In the last category, the difference (versus "adjustment obtained") constituted 44.5% of the weeks lost because of sickness absence in the last half of pregnancy. The odds ratio (OR) for obtaining job adjustment was larger for workplaces with more than 50 employees (OR 1.4) and smaller for jobs with work schedules other than daytime or shift work (OR 0.5) and also for women living with children under 16 years of age (OR 0.8). CONCLUSIONS: Job adjustment is associated with reduced sickness absence during pregnancy. Further studies should explore workplace characteristics that make it difficult to obtain such adjustments, as required by law. PMID- 9403470 TI - Sensitization to industrial enzymes in enzyme research and production. AB - OBJECTIVES: This study investigated sensitization to industrial enzymes in Finnish enzyme production and in Finnish laboratories. METHODS: A cross-sectional study was conducted in 2 plants producing industrial enzymes and in their product development and research laboratories. Sensitization to enzymes and environmental allergens was examined by skin prick tests and specific immunoglobulin E determinations (radioallergosorbent test). The employees were interviewed for work-related respiratory symptoms. Altogether 173 employees were examined. RESULTS: The skin prick test showed 21 employees (12%) to be sensitized to one or more enzymes. Sixteen positive persons also had specific immunoglobulin E. Atopy was distinctly associated with enzyme sensitization. An exposure-response relationship was found for enzyme sensitization and for respiratory symptoms during work. For sensitization, the exposure-response linear trend was statistically significant. It weakened but remained statistically significant after stratification for atopy. For symptoms, likewise, the exposure-response linear trend was statistically significant, and the statistical significance remained after stratification for atopy. CONCLUSION: The study confirmed that industrial enzymes are potent sensitizers. The handling of dry enzymes in laboratory work may cause sensitization. Sensitization may even follow minute degrees of exposure, such as among office personnel. Atopics are more susceptible to sensitization than nonatopics. Nonatopics are also clearly at risk; the demonstrated exposure-response relationship emphasizes the need for and advantages of proper exposure control. PMID- 9403471 TI - Developing national indicators for occupational health. PMID- 9403472 TI - Occupational diseases in Finland in 1996. PMID- 9403473 TI - Aging and the brain: a new frontier. AB - With aging, both "normal" senescent age-related changes (ARCs) and late-onset diseases affect the brain, producing declines in performance. The brain as a postmitotic structure is particularly vulnerable to ARCs, and senescence is by far the most powerful risk factor for neurological diseases of the elderly such as sporadic Alzheimer's disease. The concept of senescence as an immutable result of the passage of time is yielding to understanding of the biology of ARCs. Both individual and species differences in longevity illustrate the variable effects of time. Whereas human life expectancy has been extended by prevention and treatment of specific diseases, life span can be altered by modifying the processes producing ARCs. Models of prolonged life span (eg, modifications of Caenorhabditis elegans longevity genes, restricted caloric intake) demonstrate the feasibility of extending longevity throughout the phylogenetic spectrum. Both programmed and variable factors produce ARCs. Cell survival depends on a balance of opposing factors--oncogene and anti-oncogene products, cyclins, growth factors, and so on; apoptotic death results when the balance shifts. Variable factors, including accumulation of oxygen free radicals, protein conformational changes, decline in chaperone functions, and secondary loss of mitochondrial energy production, can also result in neuronal degeneration. To prevent the increased neuronal vulnerability of senescence, ARCs must be modified. The "new frontier" in neurology is the challenge of understanding the changes of aging, both to determine their impact on disease and to prevent their consequences. PMID- 9403475 TI - Inhibition of axonal growth from sensory neurons by excess nerve growth factor. AB - Fifteen-day embryonic rat dorsal root ganglion (DRG) neurons were exposed to 1 to 200 ng/ml nerve growth factor (NGF). Maximal neurite outgrowth was obtained with 10 to 20 ng/ml. Neurite outgrowth was reduced to 89% of maximal by increasing NGF to 50 ng/ml, to 66% by 100 ng/ml, and to 18% by 200 ng/ml NGF. Identical effects were seen with mouse 2.5S NGF and recombinant human NGF. Neuron cell counts demonstrated that significant cell death did not occur. In time course experiments, significant inhibition, compared with control, began within 1 hour of adding 200 ng/ml and 3 hours of adding 50 ng/ml NGF. The inhibitory effect of NGF on neurite outgrowth was reversed within 3 hours when DRG were incubated with 5 ng/ml NGF after treatment with 50 or 200 ng/ml NGF medium for 12 hours. The inhibition demonstrated for neurons did not occur in PC12 cells; axonal growth was not inhibited by up to 1,000 ng/ml NGF. Excess brain-derived neurotrophic factor or neurotrophin-3 did not inhibit neurite outgrowth. We conclude that high concentrations of NGF produces specific and reversible arrest of neurite outgrowth from sensory neurons. This observation has important clinical implications, because these inhibitory concentrations have been exceeded when NGF has been administered into the central nervous system of humans and animals. PMID- 9403474 TI - Presurgical multimodality neuroimaging in electroencephalographic lateralized temporal lobe epilepsy. AB - The purpose of this study was to compare 2-[18F]fluoro-2-deoxy-D-glucose positron emission tomography (FDG-PET), hippocampal volumetry (HV), T2 relaxometry, and proton magnetic resonance spectroscopic imaging (1H-MRSI) in the presurgical neuroimaging lateralization of patients with nonlesional, electroencephalogram (EEG)-defined unilateral temporal lobe epilepsy (TLE). Twenty-five patients were prospectively studied, along with age-matched controls. T2 relaxometry examinations were performed in 13 patients. Comparison of FDG-PET, HV, and 1H MRSI was possible in 23 patients. FDG-PET lateralized 87% of patients, HV 65%, N acetyl aspartate (NAA)/(choline [Cho] + creatine [Cr]) 61%, and [NAA] 57%. Combined HV and NAA/(Cho + Cr) results lateralized 83% of the patients, a value similar to PET. Of 10 patients with normal magnetic resonance imaging (MRI) scans, 2 were lateralized with HV, 6 with FDG-PET, 4 with NAA/(Cho + Cr), and 3 with [NAA]. T2 relaxometry lateralized no patients without hippocampal atrophy. Bilateral abnormality was present in 29 to 33% of patients with 1H-MRSI measures and 17% with HV. Only hippocampal atrophy correlated with postoperative seizure free outcome. FDG-PET remains the most sensitive imaging method to correlate with EEG-lateralized TLE. Both FDG-PET and 1H-MRSI can lateralize patients with normal MRI, but only the presence of relative unilateral hippocampal atrophy is predictive of seizure-free outcome. Bilaterally abnormal MRI and 1H-MRSI measures do not preclude good surgical outcome. PMID- 9403476 TI - Human immunodeficiency virus type 1 and its coat protein gp120 induce apoptosis and activate JNK and ERK mitogen-activated protein kinases in human neurons. AB - Detection of apoptotic neurons and microglial cells in the brains of human immunodeficiency virus type 1 (HIV-1)-infected patients has suggested that programmed cell death may be implicated in the physiopathology of HIV-1 encephalopathy. To analyze in vitro the intracellular signals induced by HIV-1 in human neurons and the associated neuronal death, we tested cultured human central nervous system (CNS) cells for apoptosis induced by HIV-1 and gp120 and for signaling pathways activated by gp120. HIV-1 and gp120 induced apoptosis of neurons and microglial cells but not of astrocytes or transformed microglial cells. Gp120 activated c-Jun N-terminal kinase (JNK) and p42 extracellular regulated kinase (ERK) in primary CNS cells, with an early peak of activation at 2 to 5 minutes that was not present when pure microglial or astrocyte cultures were tested, followed by a late and sustained activation (10 and 60 minutes) in primary and enriched glial cell cultures as well as in transformed microglial cells. This demonstrates that gp120 could be an effector of HIV-1-induced apoptosis in the CNS and act directly on neuronal and glial cells. PMID- 9403477 TI - A randomized trial of anticoagulants versus aspirin after cerebral ischemia of presumed arterial origin. The Stroke Prevention in Reversible Ischemia Trial (SPIRIT) Study Group. AB - Aspirin is only modestly effective in the secondary prevention after cerebral ischemia. Studies in other vascular disorders suggest that anticoagulant drugs in patients with cerebral ischemia of presumed arterial (noncardiac) origin might be more effective. The aim of the Stroke Prevention in Reversible Ischemia Trial (SPIRIT) therefore was to compare the efficacy and safety of 30 mg aspirin daily and oral anticoagulation (international normalized ratio [INR] 3.0-4.5). Patients referred to a neurologist in one of 58 collaborating centers because of a transient ischemic attack or minor ischemic stroke (Rankin grade < or =3) were eligible. Randomization was concealed, treatment assignment was open, and assessment of outcome events was masked. The primary measure of outcome was the composite event "death from all vascular causes, nonfatal stroke, nonfatal myocardial infarction, or nonfatal major bleeding complication." The trial was stopped at the first interim analysis. A total of 1,316 patients participated; their mean follow-up was 14 months. There was an excess of the primary outcome event in the anticoagulated group (81 of 651) versus 36 of 665 in the aspirin group (hazard ratio, 2.3; 95% confidence interval [CI], 1.6-3.5). This excess could be attributed to 53 major bleeding complications (27 intracranial; 17 fatal) during anticoagulant therapy versus 6 on aspirin (3 intracranial; 1 fatal). The bleeding incidence increased by a factor of 1.43 (95% CI, 0.96-2.13) for each 0.5 unit increase of the achieved INR. Anticoagulant therapy with an INR range of 3.0 to 4.5 in patients after cerebral ischemia of presumed arterial origin is not safe. The efficacy of a lower intensity anticoagulation regimen remains to be determined. PMID- 9403478 TI - Correlation between PMP-22 messenger RNA expression and phenotype in hereditary neuropathy with liability to pressure palsies. AB - Hereditary neuropathy with liability to pressure palsies (HNPP) is associated with a deletion in chromosome 17p11.2, which includes the gene for the peripheral myelin protein 22 (PMP-22). A "gene dosage" effect is probably the mechanism underlying HNPP, but the amount of PMP-22 mRNA in sural nerves of HNPP patients is highly variable and the role of PMP-22 underexpression in impairing myelination has yet to be clarified. We have studied 6 genetically proven HNPP patients, to evaluate the relationship between PMP-22 mRNA levels, and clinical, neurophysiological, and neuropathological findings. Underexpression of PMP-22 mRNA correlates with disease severity and with mean axon diameter and g ratio, but not with myelin thickness, number of "tomacula," or nerve conduction parameters. Our findings further confirm that underexpression of PMP-22 is the main pathogenetic mechanism underlying the severity of clinical symptoms and signs in HNPP. Smaller axons in sural nerves of HNPP patients with lower PMP-22 levels suggests that underexpression of PMP-22 may also affect axon development. PMID- 9403479 TI - Outcome after temporal lobectomy in bilateral temporal lobe epilepsy. AB - We determined how noninvasive presurgical data relate to prognosis after temporal lobectomy in patients with independent bilateral temporal lobe (IBTL) complex partial seizures on the intracranial electroencephalogram (EEG). Between 1986 and 1994, 28 patients had IBTL seizures on intracranial EEG. Fifteen of these 28 patients underwent temporal lobectomy and 13 were not offered surgery. Of the 15 patients who had surgery, 10 patients became seizure-free. Magnetic resonance imaging (MRI) and the Wada test were the only variables associated with a seizure free outcome. Seven of 10 seizure-free patients had a lateralized Wada result or the presence of unilateral hippocampal sclerosis, whereas none of the patients with persistent seizures had either of these findings. Variables not found to be predictive of a seizure-free outcome included location of scalp interictal spikes, degree of seizure-onset laterality, presence of early epilepsy risk factor, duration of epilepsy, and full-scale intelligence quotient. We conclude that MRI and the Wada test provide information of prognostic value in patients with bilateral temporal seizures independent of intracranial EEG data. PMID- 9403480 TI - Spinocerebellar ataxia type 6: CAG repeat expansion in alpha1A voltage-dependent calcium channel gene and clinical variations in Japanese population. AB - Autosomal dominant spinocerebellar ataxias (SCAs) are clinically and genetically a heterogeneous group of neurodegenerative disorders. Recently, mild CAG repeat expansion in the alpha1A voltage-dependent calcium channel gene has been found to be associated with a type of autosomal dominant SCA (SCA6). We analyzed 98 Japanese families with autosomal dominant SCAs, for whom CAG repeat expansions of the SCA1, SCA2, Machado-Joseph disease/SCA3, and dentatorubral-pallidoluysian atrophy genes were excluded, and 5 apparently sporadic cases of cortical cerebellar atrophy. The diagnosis of SCA6 was confirmed in 30 families (31%) comprising 47 affected individuals and 1 sporadic case. The size of expanded CAG repeats ranged from 21 to 26 repeat units and was found to be correlated inversely with age at onset. We identified 2 SCA6 patients homozygous for expanded CAG repeats, whose ages at onset were earlier than the 95% lower confidence level, suggesting the presence of a gene dosage effect of expanded CAG repeat. Ataxia is the most common initial symptom found in 45 of the 48 patients. Patients with a prolonged disease course showed other accompanying clinical features including dystonic postures, involuntary movements, and abnormalities in tendon reflexes. PMID- 9403481 TI - Mapping of a second locus for familial hemiplegic migraine to 1q21-q23 and evidence of further heterogeneity. AB - Familial hemiplegic migraine (FHM) is an autosomal dominant variety of migraine with aura. We previously mapped an FHM gene on the short arm of chromosome 19. Mutations in this gene, recently shown to be the alpha1 subunit of a P/Q-type voltage-dependent calcium channel, CACNL1A4, are involved in approximately 50% of unselected FHM families and in all families where migraine attacks are associated with permanent cerebellar ataxia. As a first step toward the identification of other FHM genes, we conducted a genetic linkage analysis in one large French pedigree and showed significant linkage to two microsatellite markers D1S2635 (Zmax: 3.33 at theta = 0.05) and D1S2705 (Zmax: 3.64 at theta = 0.05), establishing the existence of a second locus for FHM (FHM2) on chromosome 1q21 q23. Analysis of six additional FHM families favored linkage to this locus in two of them; linkage was excluded in the last four families, indicating further heterogeneity. Chromosome 1-linked families differ from the ones linked to chromosome 19, because penetrance in those families is much lower, and in some of their members, epileptic seizures occur during severe migraine attacks. PMID- 9403482 TI - Unilateral cerebellar lesions affect initiation of ipsilateral smooth pursuit eye movements in humans. AB - To clarify the role of the cerebellum in pursuit initiation (first 100 msec), we used infrared oculography to examine the effect of unilateral cerebellar lesions on the initial (0-20 msec) and later (80-100 msec) periods as well as the steady state response (200-300 msec) of horizontal smooth pursuit in 10 patients with unilateral cerebellar lesions. These results were compared with those of 17 age matched healthy subjects. Smooth pursuit was elicited with a step-ramp target movement with randomized horizontal directions and velocities of 10 degrees/sec and 30 degrees/sec. In the first 20-msec pursuit, velocity was 22% lower toward the side of the cerebellar lesion than away from it and 16% lower in the period 80 to 100 msec (normal differences, 2% and 3%). Later (200-300 msec), the ipsiversive/contraversive difference was smaller, but pursuit velocity in both directions was significantly lower in patients than in normals. No lesion affected the floccular region and/or the nodulus/uvula. Five lesions extended so far medially that they could have affected the medial deep cerebellar nucleus (fastigial nucleus). The remaining five were in the lateral hemisphere in areas previously considered uninvolved in pursuit generation. Our findings prove that the cerebellum participates in human pursuit initiation and that lesions in the lateral cerebellum possibly affect smooth pursuit. PMID- 9403483 TI - Amyloid beta-protein deposition in the leptomeninges and cerebral cortex. AB - To further investigate the process of amyloid beta-protein (Abeta) deposition, we determined, using sensitive enzyme immunoassays, the levels of Abeta40 and Abeta42 (Abetas) in the soluble and insoluble fractions of the leptomeninges (containing arachnoid mater and leptomeningeal vessels) and cerebral cortices from elderly control subjects showing various stages of Abeta deposition and from patients affected by Alzheimer's disease (AD). In both locations, insoluble Abeta levels were higher by orders of magnitude than soluble Abeta levels. Soluble Abeta levels in cortices were much lower than those in leptomeninges. In insoluble Abeta in the cortex, Abeta42 was by far the predominant species, and Abeta42 in AD cortices was characterized by the highest degree of modifications in the amino terminus. In contrast, this Abeta42 predominance was not observed in insoluble Abeta in the leptomeninges, which were found to be able to accumulate Abetas to an extent similar to that in the cortex, on a weight basis. The levels of insoluble Abeta in the leptomeninges or cortex generally correlated with the degree of cerebral amyloid angiopathy or the abundance of senile plaque, respectively. However, the presence of plaque-free cortical samples showing significant levels of insoluble Abeta42 suggests that biochemically detectable Abeta accumulation precedes immunocytochemically detectable Abeta deposition in the cortex. PMID- 9403484 TI - Butyrylcholinesterase in the life cycle of amyloid plaques. AB - Deposits of diffuse beta-amyloid (Abeta) may exist in the brain for many years before leading to neuritic degeneration and dementia. The factors that contribute to the putative transformation of the Abeta amyloid from a relatively inert to a pathogenic state remain unknown and may involve interactions with additional plaque constituents. Matching brain sections from 2 demented and 4 nondemented subjects were processed for the demonstration of Abeta immunoreactivity, butyrylcholinesterase (BChE) enzyme activity, and thioflavine S binding. Additional sections were processed for the concurrent demonstration of two or three of these markers. A comparative analysis of multiple cytoarchitectonic areas processed with each of these markers indicated that Abeta plaque deposits are likely to undergo three stages of maturation, ie, a "diffuse" thioflavine S negative stage, a thioflavine S-positive (ie, compact) but nonneuritic stage, and a compact neuritic stage. A multiregional analysis showed that BChE-positive plaques were not found in cytoarchitectonic areas or cortical layers that contained only the thioflavine S-negative, diffuse type of Abeta plaques. The BChE-positive plaques were found only in areas containing thioflavine S-positive compact plaques, both neuritic and nonneuritic. Within such areas, almost all (>98%) BChE-containing plaques bound thioflavine S, and almost all (93%) thioflavine S plaques contained BChE. These results suggest that BChE becomes associated with amyloid plaques at approximately the same time that the Abeta deposit assumes a compact beta-pleated conformation. BChE may therefore participate in the transformation of Abeta from an initially benign form to an eventually malignant form associated with neuritic tissue degeneration and clinical dementia. PMID- 9403485 TI - A triethnic comparison of intracerebral hemorrhage mortality in Texas. AB - Intracerebral hemorrhage (ICH) is a significant cause of stroke death. Little is known about the relative risk of Hispanic Americans (HAs), African Americans (AAs), and non-Hispanic whites (NHWs) for ICH mortality. Based on the high prevalence of hypertension in AAs and the low prevalence of hypertension in HAs, we expected AAs to have the highest ICH mortality rates and HAs the lowest. Race/ethnic age-specific ICH mortality rates were calculated from Texas vital statistics for the years 1980 through 1995. Rate ratios (RRs) are reported with NHWs as the referent group. There were 15,042 deaths due to ICH in Texas during this time. In the 45- to 59-year age group, AAs had an RR of 4. The RR for HAs was 1.9. In the 60- to 74-year age range, AAs had an RR of 1.7 and HAs had an RR of 1.3. In the 75+ age group, the rates were similar among all three race/ethnic groups. We conclude that there is a significant interaction of age and race/ethnicity for ICH. At younger ages, AAs and HAs have the highest ICH mortality rates. Access to care and socioeconomic status may play a role in the unexpectedly high ICH mortality rates in HAs. PMID- 9403486 TI - Autosomal dominant cerebellar ataxia: phenotypic differences in genetically defined subtypes? AB - Seventy-seven families with autosomal dominant cerebellar ataxia were analyzed for the CAG repeat expansions causing spinocerebellar ataxia (SCA) types 1, 2, 3, and 6. The SCA1 mutation accounted for 9%, SCA2 for 10%, SCA3 for 42%, and SCA6 for 22% of German ataxia families. Seven of 27 SCA6 patients had no family history of ataxia. Age at onset correlated inversely with repeat length in all subtypes. Yet the average effect of one CAG unit on onset age was different for each SCA subtype. We compared clinical, electrophysiological, and magnetic resonance imaging (MRI) findings to identify phenotypic characteristics of genetically defined SCA subtypes. Slow saccades, hyporeflexia, myoclonus, and action tremor proposed SCA2. SCA3 patients frequently developed diplopia, severe spasticity or pronounced peripheral neuropathy, and impaired temperature discrimination, apart from ataxia. SCA6 presented with a predominantly cerebellar syndrome and patients often had onset after 55 years of age. SCA1 was characterized by markedly prolonged peripheral and central motor conduction times in motor evoked potentials. MRI scans showed pontine and cerebellar atrophy in SCA1 and SCA2. In SCA3, enlargement of the fourth ventricle was the main sequel of atrophy. SCA6 presented with pure cerebellar atrophy on MRI. However, overlap between the four SCA subtypes was broad. PMID- 9403487 TI - Spinocerebellar ataxia type 6: gaze-evoked and vertical nystagmus, Purkinje cell degeneration, and variable age of onset. AB - Spinocerebellar ataxia type 6 (SCA6) was recently identified as a form of autosomal dominant cerebellar ataxia associated with small expansions of the trinucleotide repeat (CAG)n in the gene CACNL1A4 on chromosome 19p13, which encodes the alpha1 subunit of a P/Q-type voltage-gated calcium channel. We describe clinical, genetic, neuroimaging, neuropathological, and quantitative oculomotor studies in four kindreds with SCA6. We found strong genetic linkage of the disease to the CACNL1A4 locus and strong association with the expanded (CAG)n alleles in two large ataxia kindreds. The expanded alleles were all of a single size (repeat number) within the two large kindreds, numbering 22 and 23 repeat units. It is noteworthy that the age of onset of ataxia ranged from 24 to 63 years among all affected individuals, despite the uniform repeat number. Radiographically and pathologically, there was selective atrophy of the cerebellum and extensive loss of Purkinje cells in the cerebellar cortex. In addition, clinical and quantitative measurement of extraocular movements demonstrated a characteristic pattern of ocular motor and vestibular abnormalities, including horizontal and vertical nystagmus and an abnormal vestibulo-ocular reflex. These studies identify a distinct phenotype associated with this newly recognized form of dominant SCA. PMID- 9403488 TI - Morphometry of in vivo human white matter association pathways with diffusion weighted magnetic resonance imaging. AB - The precise characterization of cortical connectivity is important for the understanding of brain morphological and functional organization. Such connectivity is conveyed by specific pathways or tracts in the white matter. Diffusion-weighted magnetic resonance imaging detects the diffusivity of water molecules in three dimensions. Diffusivity is anisotropic in oriented tissues such as fiber tracts. In the present study, we used this method to map (in terms of orientation, location, and size) the "stem" (compact portion) of the principal association, projection, and commissural white matter pathways of the human brain in vivo, in 3 normal subjects. In addition, its use in clinical neurology is illustrated in a patient with left inferior parietal lobule embolic infarction in whom a significant reduction in relative size of the stem of the left superior longitudinal fasciculus was observed. This represents an important method for the characterization of major association pathways in the living human that are not discernible by conventional magnetic resonance imaging. In the clinical domain, this method will have a potential impact on the understanding of the diseases that involve white matter such as stroke, multiple sclerosis, amyotrophic lateral sclerosis, head injury, and spinal cord injury. PMID- 9403489 TI - Dendritic injury is a pathological substrate for human immunodeficiency virus related cognitive disorders. HNRC Group. The HIV Neurobehavioral Research Center. AB - To determine the neuropathological substrate of human immunodeficiency virus (HIV)-associated neurocognitive disorders, we examined persons with acquired immunodeficiency syndrome before their death and related their antemortem neuropsychological performance to postmortem indicators of HIV encephalitis, viral burden, and presynaptic and postsynaptic neuronal injury. Of 20 prospectively examined cases, 9 were neurocognitively normal, 5 showed neuropsychological impairment, 5 had minor cognitive/motor disorder, and 1 was demented. Degree of neurocognitive impairment was strongly related to the amount of dendritic simplification based on microtubule-associated protein 2 immunohistochemical staining, somewhat less so to a semiquantitative viral burden score based on numbers of HIV gp41-immunoreactive cells, and much less so to the presence of multinucleated giant cells or microglial nodules. It appears that even milder neurocognitive impairment reflects microneuroanatomical injury to synaptic structures. PMID- 9403490 TI - Treatment of gustatory sweating with botulinum toxin. AB - Gustatory sweating is an autonomic disorder that frequently occurs after parotid gland surgery. We investigated the action of intracutaneous injections of botulinum toxin (BTX) (1.0-2.0 mouse units/2.25-cm2 skin area) in 45 patients (mean age, 52 years) with gustatory sweating. The area of hyperhidrosis was determined by Minor's iodine test before and up to 24 weeks after the injection. The effect of BTX was assessed by measuring the hyperhidrotic area. The maximum BTX-induced reduction of gustatory sweating was seen at 7.4 +/- 4.5 days after injection. The area of sweating decreased from 17.6 +/- 8.6 cm2 before BTX to 1.3 +/- 1.6 cm2 after BTX (p < 0.0001). Half the patients rated gustatory sweating subjectively as completely abolished, and the remainder felt pronounced improvement. No toxic effects were observed. In none of the patients did hyperhidrosis recur over a 6-month follow-up. We conclude that BTX is a safe and effective treatment that can be recommended as the therapy of choice in gustatory sweating. PMID- 9403491 TI - Are portable folding chairs useful to combat orthostatic hypotension? AB - The effect of sitting on a derby chair (height, 48 cm), fishing chair (38 cm), and footstool (20 cm) on the improvement of postural hypotension was studied serially in 8 patients with autonomic failure. The increment in blood pressure during sitting was higher by using a lower portable chair. This was related to an increasing cardiac output at lower sitting heights. The fishing chair was, according to our patients' experience, most useful. PMID- 9403492 TI - Decreased cerebrospinal fluid levels of substance P in patients with Rett syndrome. AB - To clarify the mechanism of brain and spinal cord impairment in Rett syndrome (RS), we measured the cerebrospinal fluid (CSF) levels of substance P in 20 patients with RS including 16 childhood patients and 4 adult patients. Findings were compared with those obtained in age-matched controls and diseased controls. The CSF level of substance P was significantly lower in patients with RS compared with controls. The alteration in the CSF level of substance P may be related to the neurological impairment, especially autonomic dysfunction, and neuropathological involvement of dorsal root ganglia and peripheral nerve observed in RS. PMID- 9403494 TI - Application of the Poser criteria in primary progressive multiple sclerosis. PMID- 9403493 TI - Ibuprofen does not suppress active multiple sclerosis lesions on gadolinium enhanced MR images. PMID- 9403495 TI - Cortical matching of visual and vestibular 3D coordinate maps. PMID- 9403496 TI - Multiple sclerosis and retroviruses. PMID- 9403497 TI - Xenotransplantation with special reference to genetic engineering. PMID- 9403499 TI - Participation of CD14 in the phagocytosis of smooth-type Salmonella typhimurium by the macrophage-like cell line, J774.1. AB - The role of CD14 in the phagocytosis and killing of microorganisms was investigated using macrophage-like cell lines, CD14-positive J774.1 cells and CD14-negative mutant J7.DEF.3 cells derived from J744.1 cells. The cells were infected with Salmonella typhimurium organisms of the smooth (S)-form LT2, mutant rough (R)-form TV148 or Staphylococcus aureus 248betaH. At 30 or 180 min incubation, the cells were washed and disrupted. Colony-forming units (CFUs) liberated from the disrupted cells were determined by quantitative cultivation, and the phagocytic index and killing rate were calculated. Both the phagocytic index and killing rate of J774.1 cells against LT2 organisms were greater than those of J7.DEF.3 cells. However, the index and rate of J774.1 cells against TV148 and 248betaH organisms were similar to those of the J7.DEF.3 cells. The phagocytosis of LT2 organisms by J774.1 cells was partially inhibited by S-form LPS (S-LPS) and anti-CD14 antibody, but not by R-chemotype LPS (R-LPS). These results suggest that CD14 participates in the phagocytosis of S-form Salmonella. PMID- 9403498 TI - Morphological features of a filamentous phage of Vibrio cholerae O139 Bengal. AB - A filamentous phage was isolated from carrier strain AI-1841 of Vibrio cholerae 0139 Bengal and thus was termed fs phage. The phage was measured to be approximately 1 microm in length and 6 nm in width. One end of the phage was slightly tapered and had a fibrous appendage. The plaques developed on strain AI 4450 of V. cholerae 0139 were small and turbid. The phage grew in strain AI-4450 and reached a size of 10(8) to 10(9) pfu/ml at 5 hr after infection without inducing any lysis of the host bacteria. The group of phages attached on rod shaped materials like fimbriae of this bacteria, with their fibrous appendages at the pointed end, were often found in the phage-infected culture. The anti fimbrial serum effectively inhibited the infection of fs phage to the host strain AI-4450. We thus concluded that the phage can be adsorbed on fimbriae with a fibrous appendage on the pointed end of the phage filament. PMID- 9403500 TI - Epidemiological study on infectious diarrheal diseases in children in a coastal rural area of Kenya. AB - Diarrheal diseases are major causes of morbidity and mortality among children in developing countries. We have analyzed the causative agents of diarrhea in children under five years of age who resided in rural environments but attended a hospital in Malindi, a coastal town in Kenya. Bacterial diarrhea was found in 239 (27.7%) of 862 patients with diarrhea. Diarrheagenic Escherichia coli, including enteropathogenic, enterotoxigenic, and enterohaemorrhagic strains, was isolated from 119 (13.8%) patients, followed by Salmonella spp. (63 cases, 7.3%) and Shigella spp. (56 cases, 6.5%). Intestinal parasites were found in 109 (12.6%) of the patients. Entamoeba histolytica and Giardia lamblia were found in 67 (7.8%) and 42 (4.9%) of the cases, respectively. Rotavirus was found in 69 (16.1%) of 428 cases, a part of the 862 cases. Significant differences in age distribution were seen in diarrheal cases due to Campylobacter spp., G. lamblia, and rotavirus. No significant seasonal incidence of specific pathogens was found, but the number of diarrheal patients was significantly correlated to rainfall. Drinking water was contaminated with bacteria at concentrations ranging from 10(3) to 10(6) CFU/ml in 98% of the households and by coliform bacteria at concentrations of 10(2) to 10(5) CFU/ml in 72% of the households. These results suggest that the main routes of infection may be contaminated drinking water and fecal-oral transmission of enteric pathogens. Consequently, we propose that the enhancement of hygienic practice through health education is a feasible control measure of diarrhea in the study area. PMID- 9403501 TI - Modification of delivery system enhances MHC nonrestricted immunogenicity of V3 loop region of HIV-1 gp120. AB - A successful peptide vaccine for AIDS is desired to elicit T-helper and cytotoxic T lymphocyte responses besides neutralizing antibodies. The V3 loop peptide of HIV-1 has been shown to contain the principal neutralizing domain, one of the most immunodominant regions, having both B-cell and T-cell determinants. In this study, the tip of the V3 loop region was mutated from GPGR to GPGQ based on the sequence of Indian isolates (CKRKIHIGPGQAFYT). To further enhance the immunogenicity of this epitope, two delivery systems of immune stimulating complexes (ISCOMs) and liposomes were used to incorporate the peptide. Mice of differing haplotypes, H-2b, H-2d, H-2k and H-2s, showed no MHC restriction when immunized with these formulations. The IgG levels as assessed by ELISA were found to be significantly higher (P < 0.05 to P < 0.001) for even five-fold lower doses of the peptide in ISCOMs and liposomes as compared to the conventional alum-based preparation. The major subtype elicited was IgG2a/IgG2b, suggestive of a Th1-like response for all the formulations. Thus, it would appear that the same peptide incorporated in ISCOMs and liposomes selects a Th1 response and may therefore be important not only for neutralization but also for virus clearance. PMID- 9403502 TI - Characteristics of IgA anti-HIV antibodies in plasma from patients with HIV infection. AB - The concentration of total IgA and the specificity and molecular size of IgA anti human immunodeficiency virus (HIV) type-1 antibodies in plasma obtained from individuals at different stages of HIV infection were analyzed. The concentration of total IgA in the plasma was not decreased even in the late stage of HIV infection, in contrast with those of total IgG and IgM. The IgA anti-HIV antibodies differed to the IgG anti-HIV antibodies in their specificity as determined by Western blotting. The IgA antibodies mainly bind to Env glycoproteins. The IgA anti-HIV antibodies in plasma were detected between IgG and IgM by gel filtration, suggesting the presence of polymeric IgA anti-HIV antibodies. These results indicate that the production of non-specific IgA in plasma is enhanced by unknown mechanisms in every stages of HIV infection, and suggest that IgA anti-HIV antibodies in plasma which are possibly polymeric and have unique specificity may play an important role in HIV infection. PMID- 9403503 TI - Amplification of rfbE and fliC genes by polymerase chain reaction for identification and detection of Salmonella serovar Enteritidis, Dublin and Gallinarum-Pullorum. AB - Polymerase chain reaction (PCR) primers for O9 antigen (rfbE) and phase 1 flagellin antigen (fliC) were designed for the rapid identification and detection of Salmonella serovar Enteritidis and Dublin. The rfbE primer pairs selectively amplified the rfbE region of group O9 Salmonella serovars. The fliC primer pairs amplified the DNAs of g,m and g,p-type flagellar antigen; Salmonella serovar Enteritidis, Dublin, and Essen. However, DNA from flagellar-negative Salmonella serovar Gallinarum-Pullorum was also amplified. The sensitivity of PCR primer pairs was 10(4) CFU/assay by boiled DNA preparation and 10(2) CFU/assay by proteinase K-treated DNA preparation. PMID- 9403504 TI - Hyaluronidase activity in human pus from which Streptococcus intermedius was isolated. AB - Hyaluronidase (HAase) activity was detected in both a human pus sample and the culture supernatant of the only bacterial isolate from the pus, Streptococcus intermedius, using a zymographic technique. The optimum pH range for HAase activity was similar for both samples. Although the bands showing the strongest HAase activity of these samples differed from each other with respect to molecular size, both samples were equally inhibited by an antiserum raised against HAase of S. intermedius. These results suggest that S. intermedius may produce HAase in vivo as well as in vitro, and that this enzyme and/or its fragments may play an important role in host tissue degradation. PMID- 9403505 TI - Protease-induced multicell formation in Staphylococcus haemolyticus. AB - Multicellular cells were efficiently induced in Staphylococcus haemolyticus by the addition of protease to exponentially growing cultures at 30 C. Electron microscopy revealed the formation of tetrad-shaped multicells that were septated but not separated from each other. Incubation of the multicells with extract from the cells grown without protease resulted in a fourfold increase in the number of colony-forming units as compared with the untreated control. An electrophoretic analysis showed that protease caused a loss of cell wall-lytic activity of the cell, which possibly led to the formation of multicells through cessation of cross wall separation. PMID- 9403506 TI - Cloning and sequence analysis of another Shiga-like toxin IIe variant gene (slt IIera) from an Escherichia coli R107 strain isolated from rabbit. AB - An Escherichia coli R107 strain (O26 serotype) producing a Shiga-like toxin IIe variant (SLT-IIera) was isolated from the mesenteric lymph node of a freshly dead rabbit carcass. The entire structural gene for this SLT-IIera was cloned from chromosomal DNA by PCR using primers based on previously published slt-IIe sequences. Nucleotide sequence analysis indicated that the slt-IIera gene was very similar to slt-IIe (formerly called slt-IIv) from E. coli strains S1191 and 412; five and one nucleotide changes were detected in A and B subunits, respectively, which resulted in changes in amino acid sequences of the corresponding subunits by three and one residues. Recombinant SLT-IIera and SLT IIe produced using an E. coli host-vector system showed similar cytotoxicity, suggesting that the variations in the structural gene of SLT-IIera have no significant effect on cytotoxic level. PMID- 9403507 TI - Cloning and characterisation of the aroA and aroD genes of Shigella dysenteriae type 1. AB - The aroA and aroD genes from Shigella dysenteriae type 1, encoding 5 enolpyruvylshikimate 3-phosphate synthase and 3-dehydroquinase, respectively, were cloned by polymerase chain reaction (PCR). Their nucleotide sequences were determined and predicted to code for 46 kDa and 27.5 kDa proteins, respectively. Protein expressed from these genes using the minicell system, corresponded to the size of the predicted protein products. The cloned genes were shown to be functional by complementation of Escherichia coli aroA- and aroD- mutants. The predicted amino acid sequences of the cloned aroA (427 amino acids) and aroD (252 amino acids) genes of S. dysenteriae type 1 were found to be highly homologous to the corresponding genes in other bacterial species, indicating the high conservation of these housekeeping genes. The use of the cloned aroA and aroD genes in the development of a vaccine strain against S. dysenteriae is discussed. PMID- 9403508 TI - Intracytoplasmic localization of antigenic heat-stable 120- to 130-kilodalton proteins (PS120) common to spotted fever group rickettsiae demonstrated by immunoelectron microscopy. AB - Immunoelectron microscopy demonstrated antigenic heat-stable 120- to 130 kilodalton proteins (PS120) of spotted fever group (SFG) rickettsiae with antiserum against recombinant PS120 of Rickettsia japonica. In the case of R. japonica, a major part of the protein was shown to be localized outside the electron-lucent nucleoid-like region in the cytoplasm of the organisms. The other SFG rickettsiae represented a similar localization of the PS120 antigens cross reactive to that of R. japonica. On the other hand, a typhus group rickettsia demonstrated no antigens cross-reactive to the PS120 of SFG rickettsiae. PMID- 9403510 TI - Prevalence of herpes simplex virus type 1- and 2- specific antibodies among the acute, recurrent, and provoked types of female genital herpes. AB - Sixty-eight sera from the acute, recurrent, and provoked types of female genital herpes were compared for the seroprevalence of herpes simplex virus (HSV) types 1 and 2 by immunodot assay using HSV glycoprotein G. In the HSV-1-isolated patients, no HSV-2 antibodies were detected, whereas in the HSV-2-isolated patients, HSV-1 seroprevalence was 9% for the acute type, 89% for the provoked type (P < 0.005), and 55% for the recurrent type (P < 0.05). The natural history of female genital herpes and the possible protective role of pre-existing antibodies in preventing the acquisition or clinical manifestation of a subsequent HSV infection are discussed. PMID- 9403511 TI - Genotypic analysis of the 5'-untranslated region of a pestivirus strain isolated from human leucocytes. AB - The 5'-untranslated genomic region of the pestivirus strain Europa, originated in human leucocytes and previously identified as bovine diarrhea virus (BVDV), was amplified by reverse transcription-PCR and sequenced. Analyses based on primary nucleotide sequence homology and on secondary palindromic sequence structures characteristic to genotypes revealed that this human isolate should be assigned to a novel genotype of pestivirus, type Ic. This newly emerged genotype was related to, but distinguishable from the three known BVDV genotypes, Ia, Ib and II. Three other bovine field isolates of BVDV originated from Germany were also found to belong to this new genotype Ic. Within pestivirus genotype Ic strains, the overall nucleotide sequence homology was 95-96%, and 88-92%, 88-90% and 77 79% with the other BVDV genotypes Ia, Ib and II, respectively. With the strains from border disease virus (genotype III) and hog cholera virus (genotype IV), homologies were less than 75%. PMID- 9403509 TI - Direct detection by PCR of Escherichia coli O157 and enteropathogens in patients with bloody diarrhea. AB - Direct detection of Escherichia coli O157 and foodborne pathogens associated with bloody diarrhea were achieved using polymerase chain reaction (PCR) after the preparation of DNA from stool specimens using the microspin technique. PCR was compared with cultivation and toxin production tests with respect to the efficiency of detection of each pathogen; E. coli O157, Vibrio parahaemolyticus, Salmonella serovar Enteritidis and Campylobacter jejuni. Detection of some or all of the above pathogens in clinical stool specimens was achieved using PCR. The minimum number of cells required for the detection of the above pathogens by PCR was 10(1) CFUs/0.5 g of stool sample. PCR was completed within 6 hr. The above pathogens were also detected in cultivation and toxin production tests. Partial purification of the template DNA using the microspin technique was essential for the elimination of PCR inhibitors from the DNA samples. This PCR method is an accurate, easy-to-read screening method for the detection of Shiga-like toxin producing E. coli O157 and enteropathogens associated with bloody diarrhea in stool specimens. PMID- 9403512 TI - Sho-saiko-to, a traditional Kampo medicine, enhances the anti-HIV-1 activity of lamivudine (3TC) in vitro. AB - Sho-saiko-to (SST), a traditional Kampo medicine, has been examined for its inhibitory effect on human immunodeficiency virus type 1 (HIV-1) replication in peripheral blood mononuclear cells (PBMCs). SST alone moderately inhibited HIV-1 replication at a concentration of 25 microg/ml. When SST was combined with zidovudine (AZT), lamivudine (3TC) or AZT plus 3TC, SST enhanced the anti-HIV-1 activity of 3TC. In contrast, SST slightly enhanced the anti-HIV-1 activity of AZT plus 3TC but did not enhance the activity of AZT alone. These results suggest that the combination of SST and 3TC has potential as a chemotherapeutic modality of HIV-1 infection. PMID- 9403513 TI - Immune response to neutralizing epitope on human cytomegalovirus gylcoprotein B in Japanese: correlation of serologic response with HLA-type. AB - Antigenic domain 1 (AD-1), located between amino acids 608 and 625 of human cytomegalovirus (CMV) gB protein, is the major domain recognized by neutralizing antibodies. Amino acids 552 to 630 are essential for the binding of neutralizing antibodies. We developed an enzyme-linked immunosorbent assay (ELISA) to detect antibodies against a fusion protein containing amino acid residues 549 to 644 of the gB polypeptide and maltose binding protein (MBP). Of 180 seropositive samples, 106 (58.9%) showed positive immuno-reactivity against the fusion protein. None of the seronegative samples reacted with the fusion protein. Among 57 seropositive individuals typed for HLA, subjects with HLA-DR9 had a higher positive rate against the fusion protein (13/14=92.9%) than those without HLA-DR9 (25/43=58.1%). In addition, subjects with HLA-DR15 had a lower positive rate against the fusion protein (7/16=43.3%) than those without HLA-DR15 (31/41=75.6%). Mean OD values of HLA-DR15-positive individuals were significantly lower than those of HLA-DR15-negative individuals. Thus, among CMV-infected individuals, HLA-DR9 may be associated with responders for neutralizing antibodies and HLA-DR15 may be associated with non/low-responders. PMID- 9403514 TI - Language, aging, and inhibitory deficits: evaluation of a theory. AB - This article evaluates the success of Inhibitory Deficit theory in addressing two basic functions of a theory: explaining available results and predicting new findings. The review focuses on language comprehension and production, domains of cognition vulnerable to age-linked inhibitory deficits under the theory. Considerable research, however, reports remarkable age constancy in many aspects of language performance, contrary to the predictions of Inhibitory Deficit theory. For conditions that do produce age differences in language comprehension and production, evidence for inhibitory deficits is controversial at best. In predicting new findings, Inhibitory Deficit theory is constrained by lack of a well specified model, producing confusion between inhibition that occurs at a behavioral level versus a theoretical level. Modification of the theory is required to bring it in line with empirical findings on language and aging, and greater specification of underlying processes is required to reduce contradictions in predictions. PMID- 9403515 TI - Inhibition in attention and aging. AB - The literature on inhibition and aging has grown steadily in the wake of Hasher & Zacks' (1988) inhibitory deficit theory of cognitive aging. Not all of the findings support the notion of an age-related inhibitory decline, and some refinement of the theory is now required. This article has three goals: (a) to evaluate the role of inhibitory mechanisms in selective attention and aging; (b) to provide an evaluation of inhibition as a theoretical concept in theories of cognitive aging, with a specific focus on attention and aging; and (c) to consider the more general problem of evaluating progress in theory development. PMID- 9403516 TI - Cognitive gerontology and attentional inhibition: a reply to Burke and McDowd. AB - Our response to the Burke and McDowd critiques (in this issue) begins with a history of the origins of the inhibitory deficit view and of its development since 1988 as well as with an account of some particularly useful findings and of our preferred mode of theory building, which is nonformal and empirically driven. Against this background, we find many points of agreement with Burke and McDowd but also many points of disagreement. For example, we agree with Burke that many aspects of language comprehension and production are age invariant, but we disagree that all such findings count against our viewpoint. Likewise, we readily acknowledge the problems in measuring inhibition that McDowd so clearly documents, but do not feel that this is a fatal problem as long as the inhibitory deficit view continues to be viable within the basic attentional literature, continues to permit the integration of a large body of existing data, and continues to generate new predictions. PMID- 9403517 TI - Anticipated support, received support, and economic stress among older adults. AB - This study examined the interface between anticipated support, received support, recent economic stressors, and depressive symptoms in later life. A theoretical perspective was developed suggesting that received support exacerbates the effects of financial stress on depressive symptoms. However, this conceptual framework further specified that the noxious effects of economic stress are buffered or offset by anticipated support. Data from a nationwide survey of elderly people provided empirical support for both hypotheses. PMID- 9403518 TI - Predicting change in activities of daily living: a longitudinal study of the oldest old in Sweden. AB - We examined predictors of stability and decline in activities of daily living (ADLs) and mobility in a population-based sample of the oldest old. Respondents were people aged 84 to 90 living in South Central Sweden. Predictors were drawn from three domains: sociodemographic variables, vitality, and physical and psychological health. Using a logistic regression model, we sought to identify variables that were associated with changes in functioning. Over the 2-year interval, we found significant main effects for stability in ADL functioning for three variables: residential status (e.g., living in the community), subjective health, and mastery (n = 142). For mobility, we identified three variables associated with stability: lung function, subjective health, and mastery (n = 137). Over the 4-year period we found that residential status was significantly associated with stability in ADL performance (n = 89), while age, marital status, grip strength, and mastery were significant predictors for stability in mobility (n = 90). The findings can direct researchers toward interventions within particular residential environments that maintain a sense of mastery and an individual's aggressive attitude toward challenging situations. PMID- 9403519 TI - Forgetful but forgiven: how age and life style affect perceptions of memory failure. AB - Young and older perceivers read a narrative in which a forgetful young or old target was described as having either a young or old life style. Perceivers attributed memory failures more to mental difficulty for old targets but to lack of effort for young targets, regardless of life style. Life style did make a difference in perceivers' memory opinion and sympathy for the old but not for the young targets. Perceivers had a less negative memory opinion when the old target had an old rather than a young life style. Also, the old target with the old life style elicited a greater degree of sympathy in young perceivers, but a lesser degree of sympathy in older perceivers. PMID- 9403520 TI - Processing speed and memory in aging and dementia. AB - We examined the role of processing speed (PS) as a mediator of age-related and dementia-related differences in cued recall and text memory. Consistent with previous research, statistical control of PS significantly attenuated or eliminated age differences on each of the memory measures. However, age-related decline in the ability to benefit from conditions of increased encoding specificity was not mediated by PS. In contrast to the results for age effects, statistical control of PS did not significantly attenuate dementia-related memory differences, suggesting that processing speed is not an important dementia related memory impairment. The implications of these findings for interpreting residual age effects and the possible influence of preclinical dementia on studies of normal aging are discussed. PMID- 9403521 TI - Frequency discrimination vs frequency estimation: adult age differences and the effect of divided attention. AB - In this experiment we explored age differences in frequency judgment. Young and older adults studied words occurring from one to six times under divided or focused attention and then completed either a frequency discrimination or a frequency estimation test for these items. Divided attention led to poorer performance on both frequency judgment tests, suggesting that distraction during the encoding of target events results in less optimal encoding of the information that is necessary for any type of frequency judgment. Contrary to the notion that older adults encode this information more superficially than young adults, older adults were as sensitive as young adults to relative differences in the frequency of target words, and distraction did not magnify age differences for either type of frequency judgment task. On the other hand, older adults were less accurate in assigning an absolute numerical value to the frequency of the target words. Altogether, the results are consistent with the idea that the encoding and/or retrieval processes required for accurate numerical estimation of frequency suffer a larger age-related decline than do those required for accurate discrimination of relative frequency. PMID- 9403522 TI - Religion, aging, and health: current status and future prospects. PMID- 9403523 TI - Religion among disabled and nondisabled persons I: cross-sectional patterns in health practices, social activities, and well-being. AB - What is the relationship between religious involvement and functional disability among elderly people? Is being disabled different for those who frequently attend religious services? Does religious involvement have an effect on subsequent change in disability? Deriving our hypotheses from traditional theories in the sociology of religion, these questions are explored in these two related articles. Both employ data from the New Haven site of the Established Populations for the Epidemiologic Study of the Elderly (N = 2812). In the first, cross sectional correlates of religious involvement and disability are examined at the baseline of the study, including multiple indicators of health practices, social activities, and subjective well-being. We test for interactions between religious attendance and disability. Findings are (a) that religious involvement in 1982 is tied to a broad array of behavioral and psychosocial resources, (b) that these resources are associated primarily with attendance at services, and not with subjective feelings of religiousness, and (c) that some of these associations are especially pronounced among disabled respondents. PMID- 9403524 TI - Religion among disabled and nondisabled persons II: attendance at religious services as a predictor of the course of disability. AB - Does religious involvement influence changes in physical health? We perform a longitudinal analysis of the effect of religious participation on functioning over a 12-year follow-up period, in a large, prospective, representative sample of elderly persons from New Haven, Connecticut, a religiously diverse community. To examine the possibility that disability or changes in disability may be affecting religious involvement, we perform a second longitudinal analysis of changes in religious practices. Finally, we ask whether psychosocial correlates explain the effect of religious involvement on disability. Findings are (a) that attendance at services is a strong predictor of better functioning, even when intermediate changes in functioning are included, (b) that health practices, social ties, and indicators of well-being reduce, but do not eliminate these effects, and (c) that disability has minimal effects on subsequent attendance. The findings illustrate the short- and long-term importance of religious participation to the health and well-being of elderly people, and suggest a particular significance for religious participation in the lives of disabled elders. PMID- 9403525 TI - Determining the amount of help used by disabled elderly persons at home: the role of coping resources. AB - The purpose of this study is to quantify the effects of coping resources on the amount (hours) of help used by disabled elderly persons in their home. The findings are based on the 1989 National Long-Term Care Survey. The distribution of help-hours is very skewed, mirroring the skewness of limitations in physical and cognitive functioning. Controlling for these limitations, the most important coping resources are the combinations of helpers who join forces, and coresidence with a helper. The effect of helpers' networks is large and consistent across marital status and living arrangements. The networks are more extensive for married than for unmarried persons. In reference to persons who rely only on nonrelatives: (a) a network of a spouse and children enables a married person to have 40 additional weekly hours of help, (b) a network of children and others enables an unmarried person to have 29 additional help-hours per week if he/she coresides with other adults and 10 additional weekly help-hours if he/she does not. The issue of concern for public policy is whether such family networks will be preserved and, if not, how to obtain the funds for the alternative of sufficient paid help in the community. PMID- 9403526 TI - Use of medical care by African American and White older persons: comparative analysis of three national data sets. AB - Historically, there has been a large gap between African Americans and Whites in access to health care, but this gap was ostensibly lessened by the advent of Medicare and Medicaid for older adults in the mid 1960s. The extent to which older African Americans continue to receive less access to medical care as a result of economic inequalities, institutionalized forms of discrimination, and life-style factors remains a subject of policy debate. Empirical enquiry has produced inconsistent results. The purpose of this study is to test the same set of models of medical use using identically measured predictor variables in three nationally representative data sets of older Americans: 1984 Study of Aging (SOA); 1984 National Long-Term Care Survey (NLTC); and the 1987 National Medical Care Expenditure Survey (NMES). Multivariate logistic regression of use of physician and hospital services and Poisson regression of amount of service use identified inconsistent results in race differences across data sets, but consistent results in terms of the importance of health status and insurance as predictors of use and amount of use. The findings suggest that health status and financial resources may be more relevant areas for policy interventions than considerations related to race and ethnicity. PMID- 9403527 TI - Differences by race in the decline of health over time. AB - Previous research on race differences in health, we believe, has failed to take into account the initial state of health of the respondents. Other research has demonstrated that elders in poor health are more likely to experience a change in their health over time. It is unclear if the greater probability of decline in health observed among African Americans is a result of being more likely to begin such observations in health states that are worse than those for Whites. This investigation examines declines in health over a 30-month period in a sample of African American and White elders who began the study in similar "good health." Findings support the supposition that African Americans are more likely to report a decline in their health, regardless of the health measure used. Differences by race in the decline of health appear to be a consequence of economic and educational discrepancies between the two groups. PMID- 9403528 TI - Dysregulated expression of neutrophil apoptosis in the systemic inflammatory response syndrome. AB - OBJECTIVE: To study the effect of the systemic inflammatory response syndrome (SIRS) or major elective surgery on the apoptosis of circulating polymorphonuclear neutrophils because an activated inflammatory response is terminated, in part, through the programmed cell death, or apoptosis, of its effector cells. DESIGN: A prospective inception cohort study. SETTING: A mixed surgical and medical intensive care unit of an adult tertiary care hospital. PATIENTS: Sixteen patients with SIRS, 7 uninfected patients who had undergone elective aortic aneurysmectomy, and 8 healthy laboratory control subjects. INTERVENTIONS: Serial blood samples were drawn for evaluation of neutrophil apoptosis, activational state, and surface receptor expression by flow cytometry. MAIN OUTCOME MEASURES: Spontaneous apoptosis was significantly delayed in neutrophils from patients with SIRS (8.6%+/-6.8%) and patients who had undergone elective aortic aneurysmectomy (11.0%+/-5.0%) when compared with controls (34.9%+/-6.8%). These neutrophils were activated as evidenced by enhanced respiratory burst activity and augmented surface expression of CD11b. Apoptosis in response to engagement of cell surface Fas (also known as CD95 or APO-1) with an agonistic antibody was blunted. Plasma from patients with SIRS or patients who had undergone elective aortic aneurysmectomy suppressed the apoptotic responses of control neutrophils (plasma from patients with SIRS, 18.8%+/-10.3%; plasma from patients who had undergone elective aortic aneurysmectomy, 20.0%+/-6.1%; P<.01). Western blot analysis showed normal expression of the key proapoptotic proteases, interleukin 1beta converting enzyme and CPP32 (also known as YAMA, apopain, and caspase 3), indicating that delayed apoptosis was not a consequence of decreased levels of proapoptotic enzymes. CONCLUSIONS: Circulating neutrophils from patients with SIRS or from patients who have undergone major elective surgery show delayed expression of constitutive programmed cell death, and antiapoptotic factors are present in the general circulation. While prolonged neutrophil survival may represent an appropriate adaptive response to injury, the presence of activated and apoptosis-resistant cells in an antiapoptotic environment may contribute to the systemic inflammatory injury characteristic of SIRS and predispose to the development of the multiple organ dysfunction syndrome. PMID- 9403529 TI - Intestinal epithelial cell regulation of macrophage and lymphocyte interleukin 10 expression. AB - BACKGROUND: The intestinal mucosa is subject to daily antigenic challenge and to injury by proinflammatory cytokines. Interleukin 10 (IL-10) is an important anti inflammatory cytokine produced by macrophages and lymphocytes that modulates this response. OBJECTIVE: To investigate the hypothesis that intestinal epithelial cells are a significant local source of IL-10 in the gut milieu and also participate in the regulation of macrophage and lymphocyte IL-10 expression in the intestinal microenvironment. METHODS: C-205 cells, a human intestinal epithelial cell line, were cultured for 2 days; lipopolysaccharide or tumor necrosis factor was then added. Media and cells were harvested at specific time points to determine the kinetics of IL-10 expression. C-205 cells were then cocultured with macrophages or lymphocytes isolated from human peripheral blood mononuclear cells, and IL-10 expression was assessed in unstimulated and stimulated conditions. Interleukin 10 protein was measured by enzyme-linked immunosorbent assay; IL-10 gene expression was measured by reverse transcriptase polymerase chain reaction. RESULTS: Constitutive production of IL-10 protein by C 205 cells was maximal at 3 days, paralleled by a peak in IL-10 messenger RNA (mRNA) expression at 24 hours. Lipopolysaccharide or tumor necrosis factor strikingly up-regulated IL-10 mRNA and protein expression by C-205 cells. Coculture of C-205 cells with macrophages or lymphocytes significantly increased lipopolysaccharide-stimulated IL-10 protein and mRNA release compared with C-205 cells, macrophages, or lymphocytes cultured alone. CONCLUSIONS: Enterocytes are a responsive source of IL-10 and may play a role in modulating production of this important cytokine by the local inflammatory cells of the gut. These redundant mechanisms to augment IL-10 production suggest a central role for this cytokine in regulation of the local intestinal inflammatory response. PMID- 9403530 TI - Induction of heat shock protein 70 protects thymocytes against radiation-induced apoptosis. AB - OBJECTIVES: To determine if induction of heat shock protein 70 (HSP 70), a stress protein that plays a cytoprotective role and inhibits cell death in response to various stimuli, will protect thymocytes and T-cell clones from radiation-induced apoptosis, and to define the mechanism of such protection. DESIGN: Thymocytes from BALB/c mice or T-lymphocyte clones were incubated at 43 degrees C for 1 hour to induce HSP 70, then irradiated. Control cells were irradiated but not heated. Fragmentation of DNA was quantitated, and p53, bax, and bcl-2 expression was analyzed at various times by the Western blot method. RESULTS: Only heated cells expressed HSP 70. The induction of HSP 70 increased basal apoptosis but significantly decreased radiation-induced apoptosis. Furthermore, introduction of an HSP 70 antisense oligomer prior to heating reversed the protective effect of HSP 70. Induction of HSP 70 in T-cell clones with sodium arsenite had a similar protective effect against radiation-induced apoptosis. Irradiation induced p53 and markedly up-regulated bax. The expression of p53 peaked at 4 hours and preceded maximal bax induction. Induction of HSP 70 prior to irradiation suppressed p53 and significantly decreased bax levels. Levels of bcl-2 were unaffected. CONCLUSIONS: Our data show that HSP 70 induction protects thymocytes from radiation-induced apoptosis by down-regulating p53 and bax expression. The induction of HSP 70 may represent a novel mechanism by which the immunosuppressive effects and the associated infectious complications of radiation therapy can be minimized. PMID- 9403531 TI - Heat shock induces IkappaB-alpha and prevents stress-induced endothelial cell apoptosis. AB - OBJECTIVE: To determine whether prior heat shock would attenuate endothelial cell apoptosis and whether any effect of preemptive heat shock is mediated through a nuclear factor kappa B and inhibitor kappa B alpha mechanism. DESIGN: A randomized, controlled in vitro study. SETTING: A laboratory in a large, academic medical center. INTERVENTIONS: Cultured primary porcine endothelial cells were treated with increasing doses of sodium arsenite (40-160 micromol/L), after which the interval until subsequent apoptotic (lipopolysaccharide-arsenite) challenge was varied (4-16 hours). The degree of cell death and apoptosis were determined using neutral red uptake and staining with annexin V and propidium iodide, respectively. Inducible heat shock protein 70 and inhibitor kappa B alpha levels in treated cells were determined by Western blot analysis. Lipopolysaccharide induced nuclear factor kappa B activity was assessed using an electrophoretic mobility shift assay. RESULTS: Prior arsenite treatment decreased cell death by apoptosis in a time- and dose-dependent manner. Specifically, a higher sodium arsenite concentration and shorter intervals afforded better protection (P=.01, 160 micromol/L at 4 hours). Protection against apoptosis correlated with increased heat shock protein 70 and inhibitor kappa B alpha levels and decreased nuclear factor kappa B binding activity. CONCLUSIONS: Arsenite, an inducer of the heat shock response, decreased stress-induced endothelial cell apoptosis. The mechanism of this protection may include decreased nuclear factor kappa B activity or increased inducible heat shock protein 70 levels. Heat shock protein 70 may serve as a molecular marker to determine not only the phenotypic state of the cell but also the durability of protection afforded by heat shock. These data support the hypothesis that stress-induced changes in transcription factor activity and protein expression can regulate the induction of apoptosis. PMID- 9403532 TI - Complement C3 production in human intestinal epithelial cells is regulated by interleukin 1beta and tumor necrosis factor alpha. AB - BACKGROUND: Sepsis and endotoxemia are associated with increased mucosal production of complement component C3; the enterocyte may be a source of C3 in these conditions. OBJECTIVE: To test the hypothesis that interleukin 1beta (IL 1beta) and tumor necrosis factor alpha (TNF-alpha) regulate the production of C3 in the enterocyte at the transcriptional level and that this regulation is potentiated by interferon gamma (IFN-gamma). METHODS: Cultured Caco-2 cells, a human intestinal epithelial cell line, were treated with various concentrations of human recombinant IL-1beta (0.005-1.25 ng/mL) or TNF-alpha (1-1000 U/mL) with or without the addition of IFN-gamma (250 U/mL). C3 levels in the culture medium were measured by enzyme-linked immunosorbent assay and cellular messenger RNA levels by Northern blot analysis. RESULTS: Treatment of the Caco-2 cells with IL 1beta or TNF-alpha resulted in a time- and dose-dependent increase in C3 production. The use of IFN-gamma alone did not affect C3 production but potentiated the effect of IL-1beta and TNF-alpha in a synergistic manner. C3 messenger RNA levels were increased following stimulation with either cytokine. CONCLUSIONS: C3 production in the enterocyte is regulated by IL-1beta and TNF alpha at the transcriptional level, and this response is potentiated by IFN gamma. The results suggest that C3 production in the intestinal mucosa may be regulated locally by cytokines in a paracrine or autocrine manner. PMID- 9403533 TI - A randomized, double-blind clinical trial comparing cefepime plus metronidazole with imipenem-cilastatin in the treatment of complicated intra-abdominal infections. Cefepime Intra-abdominal Infection Study Group. AB - OBJECTIVE: To evaluate the safety and efficacy of cefepime hydrochloride plus metronidazole vs the combination of imipenem and cilastatin sodium in the treatment of complicated intra-abdominal infections in adult patients. DESIGN: Prospective, randomized, double-blind multicenter study. SETTING: University affiliated hospitals in the United States and Canada. PATIENTS: Three hundred twenty-three patients with complicated intra-abdominal infections in whom an operative procedure or percutaneous drainage was required for diagnosis and management. INTERVENTION: Cefepime, 2 g, was administered intravenously every 12 hours (n= 164) in addition to metronidazole, 500 mg (or 7.5 mg/kg) intravenously every 6 hours. Imipenen-cilastatin sodium, 500 mg, was administered intravenously every 6 hours (n= 159). Surgical infection management was determined by the patients' surgeons. MAIN OUTCOME ASSESSMENTS: Clinical cure, defined as elimination of all signs and symptoms relevant to the original infection; and treatment failure, defined as persistence, increase or worsening of signs and symptoms resulting in an antibiotic change, requirement of an additional surgical procedure to cure the infection, or a wound infection with fever. RESULTS: Of the initial isolates, 84% were susceptible to cefepime and 92% were susceptible to imipenem-cilastatin. Among the 217 protocol-valid patients, those treated with cefepime+metronidizole were deemed clinical cures (88%) more frequently than were imipenem-cilastatin-treated patients (76%) (P=.02). Using multivariate analysis to adjust for identified clinical risk factors for an adverse outcome (severity of presenting illness, isolation of enterococcus, type of infection, and duration of prestudy hospitalization), there was a trend (P=.06) toward a higher cure rate favoring cefepime+metronidazole. Pathogens were eradicated in significantly (P=.01) more patients treated with combined cefepime and metronidazole (89%) than with imipenem-cilastatin (76%). CONCLUSION: The combination of cefepime plus metronidazole is safe and effective therapy for patients with severe intra abdominal infections. PMID- 9403535 TI - Enteral feeding intolerance: an indicator of sepsis-associated mortality in burned children. AB - OBJECTIVE: To determine if enteral feeding intolerance (EFI) is associated with sepsis and increased mortality in children with severe burns. DESIGN: A survey. SETTING: A pediatric burn unit. PATIENTS: Ninety-one children surviving longer than 5 days with greater than 80% total body surface area burns. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Enteral feeding intolerance indicated by high gastric residuals (> 150 mL/h) or uncontrollable diarrhea (> 2500 mL/d); thrombocytopenia (platelet count < 100 x 10(9)/L); hyperglycemia (glucose level > 11.1 mmol/L [> 200 mg/dL]); sepsis (pathogenic bacteremia or fungemia noted on blood culture results); and mortality. RESULTS: Neither EFI nor sepsis developed in 71 patients, EFI alone developed in 2 patients, sepsis alone developed in 5 patients, and EFI and sepsis developed in 13 patients. Enteral feeding intolerance and sepsis were associated by contingency table analysis (P<.001). Mortality was 8% (6 patients) in those with neither EFI nor sepsis, 50% (1 patient) in those with EFI alone, 60% (3 patients) in those with sepsis alone, and 77% (10 patients) in those with EFI-associated sepsis. The 2 latter groups were different from the group with neither EFI nor sepsis (P<.05). Enteral feeding intolerance was identified in 70% of patients before sepsis; thrombocytopenia, 64%; and hyperglycemia, 66%. When compared with thrombocytopenia and hyperthermia, no variables were found to be superior to others for predicting sepsis. CONCLUSIONS: Enteral feeding intolerance was associated with the development of sepsis and increased mortality in children with greater than 80% total body surface area burns. This sign was identified in 70% of the cases before pathogens were found in the blood; no difference could be shown between the identification of EFI, thrombocytopenia, and hyperglycemia before sepsis. These data indicate that the development of EFI should be used as an indicator of infection and should prompt a search for an inciting focus. PMID- 9403534 TI - A temporal study of TPN-induced changes in gut-associated lymphoid tissue and mucosal immunity. AB - BACKGROUND: Total parenteral nutrition (TPN) is associated with decreases in small-intestinal gut-associated lymphoid tissue (GALT) T cells, B cells, and IgA levels and impairs IgA-mediated defenses in the respiratory tract. The impaired respiratory tract defenses are speculated to be due to reduced respiratory tract IgA levels. OBJECTIVES: To determine the time course of GALT cell reductions and document any changes in respiratory tract IgA levels in mice receiving TPN. DESIGN: Prospective randomized trial. SETTING: Animal research laboratory. MATERIALS: Thirty-five male ICR mice weighing 25 to 35 g. INTERVENTIONS: Mice underwent cannulation with intravenous catheters and received chow for 2 days followed by TPN for 0 (n=6), 1 (n=6), 2 (n=6), 3 (n=6), 4 (n=6), or 5 (n=5) days. Mice were killed after receiving TPN their respective number of days. The small intestine was harvested, and washings were obtained from the small intestine and the respiratory tract. Lymphocytes and IgA levels were analyzed by flow cytometry and enzyme-linked immunosorbent assay, respectively. MAIN OUTCOME MEASURES: Lymphocyte yields from Peyer patches, intraepithelial spaces, and the lamina propria; IgA levels from the small intestine and the respiratory tract. RESULTS: T- and B-cell yields in the Peyer patches and lamina propria were significantly reduced by day 2 (P<.05) and thereafter compared with day 0. The lamina propria CD4+/CD8+ ratio declined significantly by day 4 (P<.05) compared with day 0. Small-intestinal and respiratory tract IgA levels were significantly diminished by day 3 (P<.05) and thereafter compared with day 0. CONCLUSION: Total parenteral nutrition produces rapid changes in GALT cell profiles and reduces respiratory tract IgA levels consistent with the impairment of respiratory IgA-mediated defenses. PMID- 9403536 TI - Surgical wound infection in renal transplantation: outcome data in 102 consecutive patients without perioperative systemic antibiotic coverage. AB - BACKGROUND: The incidence of surgical wound infection in the presence of immunosuppression has been reported in the literature to approach 7%. Perioperative systemic antibiotic therapy is routinely used to reduce the occurrence of wound infections. This therapy is not without complications, including adverse effects and development of resistant strains. DESIGN: Surgical wound infection rates during the first 100 days after renal transplantation were studied in 102 consecutive patients. Eighty-one patients underwent cadaveric transplantation and 21 patients underwent living-related donor transplantation from February 1, 1991, to January 1, 1992. No systemic perioperative antibiotic coverage was used, but local antibiotic irrigation was part of the perioperative protocol. SETTING: Hahnemann University Hospital, Philadelphia, Pa, is a large, tertiary care center. Patients were initially hospitalized and were discharged during the 100-day follow-up period based on clinical status and improvement in renal function. PATIENTS: Twenty-seven (25%) of 102 patients had diabetes mellitus. INTERVENTIONS: Induction immunosuppression consisted of azathioprine, prednisone, and anitlymphocyte globulin, while maintenance immunosuppression consisted of azathioprine, prednisone, and cyclosporine. Acute allograft rejection episodes were treated with steroids and/or OKT3 (Ortho Pharmaceutical Group, Raritan, NJ). RESULTS: Two surgical wound infections (2%) occurred. In both, infection was superficial, resolving with wound drainage and intravenous antibiotics. The surgical wound infection rate was not significantly affected by age, sex, allograft source, or presence of diabetes mellitus. CONCLUSIONS: Despite immunosuppression, the incidence of surgical wound infection was minimal, comparing favorably to rates reported for renal transplantation with the use of systemic antibiotics. Possible explanations for the low incidence of surgical wound infections include local wound irrigation, meticulous hemostasis, improved organ procurement techniques, and continuity in perioperative care. PMID- 9403537 TI - Influences of type and duration of antimicrobial prophylaxis on an outbreak of methicillin-resistant Staphylococcus aureus and on the incidence of wound infection. AB - OBJECTIVE: To clarify how antibiotic prophylaxis influenced an outbreak of methicillin-resistant Staphylococcus aureus (MRSA) and postoperative infection. DESIGN: Retrospective review. SETTING: University-affiliated teaching hospital. PATIENTS: All patients (n=1824) undergoing subtotal esophagectomy, gastrectomy, or colorectal surgery during the period 1982 through 1995. MAIN OUTCOME MEASURES: Type, timing, and duration of prophylactic antibiotics. Postoperative infection by the Centers for Disease Control and Prevention definition and the organisms isolated. RESULTS: Third-generation cephalosporins were frequently administered for prophylaxis during the period 1982 through 1990. The rate of isolates of MRSA from the infected site increased, peaking in 1988 to 1990. Since 1991 to 1992, along with a marked decrease in third-generation cephalosporin use, the rates of MRSA isolated have declined dramatically. The timing of administration changed from postoperative to intraoperative. Although the duration was gradually decreased, coverage was still provided until about the fifth postoperative day, even during 1993 to 1995. Prolonged coverage did not reduce the rate of superficial incisional or organ/space surgical site infection or that of pneumonia. CONCLUSIONS: Overuse of third-generation cephalosporins for long periods caused an MRSA outbreak. Long-term prophylaxis did not lower infection rates. The briefest possible prophylaxis with first- or second-generation cephalosporins should be used in general surgery. PMID- 9403538 TI - Suppression of natural killer cell activity in patients with fracture/soft tissue injury. AB - BACKGROUND: Natural killer cells (NKCs) participate in "innate" cell-mediated immunity. Fracture/soft tissue injuries are cytokine rich and may influence cell mediated immunity. OBJECTIVE: To study the effects of fracture cytokines on NKC function. DESIGN: A case-control study. SETTING: A level I trauma center and laboratory in a university medical center. PARTICIPANTS: Patients requiring open fracture fixation and healthy volunteers. INTERVENTIONS: Fracture supernatants and peripheral plasma were collected during open fracture fixation. Volunteer mononuclear cells were used as effector (NKC) sources. Mononuclear cells were preincubated with fracture supernatants, paired peripheral plasma, or normal plasma under various conditions. MAIN OUTCOME MEASURES: Natural killer cell lysis of K562 target cells was assessed by chromium 51 release. RESULTS: Fracture supernatants suppressed NKC function more rapidly than peripheral plasma. Fracture supernatants from 1 to 4 days after injury were most suppressive. Inactivation of complement and reactive oxygen species failed to restore lysis. Neutralizing antibodies to interleukin 4 and interleukin 10 further suppressed lysis. Antibodies to transforming growth factor beta1 failed to restore lysis. The addition of interferon gamma did not restore lysis but the addition of interleukin 12 did. CONCLUSIONS: Fracture supernatants and peripheral plasma from patients with fractures suppress NKCs. The responsible mediators may be concentrated in fracture/soft tissue injuries. Responses to manipulation of the cytokine environment suggest that fracture cytokines may impair cooperation between NKCs and accessory cells. PMID- 9403539 TI - Splenic abscess: another look at an old disease. AB - OBJECTIVE: To study the changes in the incidence, causes, bacteriologic profile, and management of a splenic abscess. DESIGN: Retrospective case study. SETTING: Tertiary, university referral center. PATIENTS: Thirty-nine patients with a splenic abscess. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Demographics, signs and symptoms, causes, risk factors, diagnostic methods, bacteriologic profile, treatment, and outcome. RESULTS: Patients presented at a mean age of 43 years (range, 2-83 years), after a mean symptomatic period of 16 days, with fever (69%), abdominal pain (56%), nausea and vomiting (38%), and splenomegaly (31%). The majority of abscesses represented metastatic infection (n=19), and 11 were secondary to immunosuppression. Twelve patients had human immunodeficiency virus disease and 9 used intravenous drugs. In patients who underwent computed tomography, all had abnormal scans (n=33), with a well-defined abscess(es) in 28. Nine abscesses were polymicrobial; monomicrobial isolates included gram-positive organisms (23%), gram-negative organisms (31%), fungi (23%), and mycobacteria (23%). Patients presenting before 1989 (1981-1988) (n=15) and those presenting after 1989 (1989-1996) (n=24) differed in risk factors (intravenous drug abuse, 0% vs 47% [P=.02]; hematologic malignancy, 43% vs 9% [P=.04]) and gram-positive isolates (18% vs 64%; P=.06). Patients underwent splenectomy (n=18), open drainage (n=4), medical therapy (n=10), or percutaneous drainage (n=5) with respective survival rates of 94%, 50%, 70%, and 100%. CONCLUSIONS: In 1996, splenic abscesses are increasingly common. Intravenous drug abuse and human immunodeficiency virus disease are significant risk factors, and the diagnosis should be considered in a patient with fever and abdominal pain who uses intravenous drugs. Antimicrobial agents should be broad since 36% of abscesses were polymicrobial, and should include coverage of gram-positive organisms. PMID- 9403540 TI - Enteral vitamin E supplementation inhibits the cytokine response to endotoxin. AB - OBJECTIVE: To evaluate the effect of short-term, high-dose enteral supplementation of 3 different vitamin E derivatives: free alpha-tocopherol (VE), alpha-tocopherol succinate (VES), and alpha-tocopherol acetate (VEA) on macrophage and monocyte activation. DESIGN: Sprague-Dawley rats (weight, 150-200 g) were assigned to 1 of 5 experimental groups: saline (control), ethanol (control), VES (100 mg/kg), VEA (100 mg/kg), or VE (100 mg/kg). Rats underwent oral gavage once per day for 5 days with 0.5 mL of their assigned solution. All vitamin E derivatives were diluted in 75% ethanol. Rats were then killed and whole-blood and peritoneal macrophages were harvested and stimulated with lipopolysaccharide (10 microg/mL) in vitro. Tumor necrosis factor (TNF) production was measured by enzyme-linked immunosorbent assay. Additional serum samples were analyzed for alpha-tocopherol concentration by high-performance lipid chromatography. RESULTS: Whole-blood TNF production was maximal in the control groups after 3 hours of incubation and began to decline by 6 hours. Supplementation with all 3 vitamin E derivatives resulted in suppression of lipopolysaccharide-induced TNF production at both time points when compared with both ethanol and saline controls (P<.05, analysis of variance [ANOVA]). All 3 vitamin E derivatives also resulted in significant inhibition of lipopolysaccharide-induced TNF production by peritoneal macrophages when compared with their ethanol-carrier control but not with the saline control (P<.05, ANOVA). The degree of TNF suppression correlated directly with serum alpha tocopherol levels. CONCLUSIONS: Our data demonstrate that a short-term, high-dose enteral supplementation of vitamin E can modulate the monocyte and macrophage response to endotoxin. These data, along with other animal studies showing a protective effect of vitamin E treatment in sepsis and ischemia-reperfusion injury, suggest a potential role for vitamin E supplementation in patients at risk of the systemic inflammatory response syndrome. PMID- 9403541 TI - Endotoxin-induced macrophage gene expression depends on platelet-activating factor. AB - BACKGROUND: The development of multiple organ failure in septic patients is due to a systemic inflammation orchestrated by macrophages (Mphi). Elucidation and control of the mechanism involved in Mphi activation in sepsis is crucial to improving survival. An early event of Mphi activation involves the hydrolysis of membrane phospholipid by phospholipase A2 (PLA2) and subsequent generation of platelet-activating factor (PAF). OBJECTIVE: We designed this study to test the hypothesis that Mphi gene expression depends on PAF. DESIGN: Rabbit alveolar Mphi were obtained by bronchoalveolar lavage and were stimulated with 10 ng/mL of Escherichia coli endotoxin lipopolysaccharide (LPS), PAF (1 micromol/L), LPS+/ CV3988 (10 micromol/L), a PAF receptor antagonist, or LPS+/-PLA2 inhibitors: AACOCF3 (50 micromol/L) or manoalide (10 micromol/L). After 4 hours of incubation, Mphi tumor necrosis factor (TNF) messenger RNA (mRNA) expression was assessed by Northern blot analyses. The TNF production in the Mphi supernatant was measured by L929 bioassays. RESULTS: The LPS-stimulated Mphi expressed increased levels of TNF mRNA and produced an enormous amount of TNF. CV3988, a PAF antagonist, inhibited LPS-induced TNF mRNA. Furthermore, inhibiting PAF production with AACOCF3, or manoalide, also inhibited LPS-induced Mphi TNF mRNA expression. The effect of PAF depends on changes in intracellular calcium concentration. Inhibitors of calcium flux attenuated the PAF effects on LPS stimulated Mphi. CONCLUSIONS: Our data suggest that LPS-induced Mphi gene expression is mediated by PAF. It is likely that modulation of PAF production or activity may be beneficial in down-regulating the overactivity of Mphi in sepsis. PMID- 9403542 TI - A prospective randomized trial of an antibiotic- and antiseptic-coated central venous catheter in the prevention of catheter-related infections. AB - OBJECTIVE: To test the efficacy of the ARROWgard (Arrow International Inc, Reading, Pa) central venous catheter (CVC) coated with silver sulfadiazine and chlorhexidine (A-CVC) in the prevention of CVC-related infections. DESIGN: Prospective, randomized trial. SETTING: A tertiary care medical center. PATIENTS AND INTERVENTION: Two hundred eighty-two patients who required CVC placement were evaluated in this study. Patients were prospectively randomized to receive either a standard CVC (S-CVC) or the A-CVC. Only fresh-stick double- and triple-lumen catheters were studied. MAIN OUTCOME MEASURES: Patients were evaluated for catheter site inflammation, catheter site colonization, local catheter-related infection, and catheter-related septicemia. RESULTS: The 2 groups were matched for age, percentage in the intensive care unit, percentage receiving total parenteral nutrition, percentage with triple-lumen catheters, and duration of catheterization. Rates of catheter site inflammation in the 2 groups were similar (12% vs 10%, S-CVC group and A-CVC group, respectively). The A-CVC was associated with a significantly decreased catheter site colonization rate (49% vs 28%; 43% reduction; P<.001) and local catheter-related infection rate (22.4% vs 5.8%; 74% reduction; P<.001). Rates of catheter-related septicemia were reduced by 41% in the A-CVC group (6.4% vs 3.8%, S-CVC group and A-CVC group, respectively), but this was not statistically significant. CONCLUSIONS: Despite a marked decrease in catheter site colonization and catheter-related infection rates, the A-CVC did not significantly reduce the incidence of catheter-related septicemia. This may be due to a greater pathogenic dependence on catheter hub contamination rather than catheter site colonization or local catheter-related infection, or the relatively short (5.2 days) duration of catheterization in this study. PMID- 9403544 TI - Willy Meyer's radical mastectomy. PMID- 9403543 TI - Carbon monoxide contributes to the cytokine-induced inhibition of surfactant synthesis by human type II pneumocytes. AB - BACKGROUND: An increase in cyclic guanosine 3',5'-monophosphate (cGMP) due to nitric oxide generation is known to participate in the mediation of the tumor necrosis factor alpha (TNF-alpha) effect in type II cells. Because guanylyl cyclase can be activated also by carbon monoxide (CO), in this study we examined the ability of human type II pneumocytes to produce CO in the presence of cytokines and the relative contribution of this molecule to the TNF-alpha and interleukin 1 effects. DESIGN: Type II pneumocytes were isolated from cadaveric multiple-organ donors by enzymatic digestion, adherence separation of macrophages, and gradient purification. After preculture for 24 hours, cells were cultured for 24 hours in the presence or absence of TNF-alpha, interleukin 1, sodium nitroprusside, N(omega)-nitro-1-arginine, CO, hemin, zinc-protoporphyrin type IX, deferoxamine mesylate, S-adenosyl-L-methionine, alpha-tocopherol, methylene blue (a guanylyl cyclase inhibitor), 8-bromine-cGMP, and combinations of these reagents. Both CO (picomole per microgram of protein) and nitric oxide release to the medium and the cGMP (picomole per microgram of protein) content of the cells were measured. In a different set of experiments, D-glucose labeled with radioactive carbon (14C) was added to the medium, and the labeling of several lipid fractions was determined (picomole per microgram of protein). RESULTS: D-[14C]glucose incorporation into phosphatidylcholine, the main surfactant component, was selectively inhibited in the presence of cytokines, CO, sodium nitroprusside, or 8-bromine-cGMP. The inhibitory effect of TNF-alpha was partially reversed by N(omega)-nitro-L-arginine, deferoxamine, or alpha tocopherol and totally reversed by methylene blue. Tumor necrosis factor alpha induced an increase in cGMP cell content and in the CO and nitric oxide release to the medium. Hemin increased CO and cGMP production and decreased phosphatidylcholine synthesis. Zinc-protoporphyrin type IX, an inhibitor of heme oxygenase, and all 3 antioxidants, which inhibited CO production, also antagonized the TNF-alpha effect on cGMP and phosphatidylcholine synthesis. CONCLUSIONS: Intracellular cGMP increase due to an endogenous generation of both CO and nitric oxide mediates the cytokine-induced inhibition of surfactant synthesis by type II pneumocytes. Both lipid peroxidation and heme oxygenase activity are sources for the observed CO production. PMID- 9403545 TI - The sixth report of the Joint National Committee: an appropriate celebration of the 25th anniversary of the National High Blood Pressure Education Program. PMID- 9403546 TI - A history of the Council for High Blood Pressure Research: the first 50 years. PMID- 9403547 TI - Theodore Cooper Memorial Lecture. A mouse view of hypertension. AB - Essential hypertension probably results from combinations of genetic variations, not necessarily the same in all afflicted persons, which individually may not cause sufficient deviation from normality to be significantly harmful. Genes contributing to hypertension are being sought by analytic experiments aimed at identifying candidate genes associated or segregating with the phenotype in humans and animals and by synthetic experiments in which changes are made in candidate genes in animals and their effects on blood pressure are determined. We have used gene targeting to vary the amounts of angiotensinogen and angiotensin converting enzyme (ACE) synthesized from their genes (Agt and Ace). These "gene titration" experiments establish that changes in Agt gene expression cause changes in the blood pressures of mice. Surprisingly, quantitative changes in Ace gene expression over a threefold range do not affect blood pressures. Computer simulations with a simple version of the renin-angiotensin system predict that changes in Agt function alter the steady state levels of both angiotensin I (Ang I) and angiotensin II (Ang II). In contrast, modest changes in Ace function alter Ang I levels considerably but scarcely affect Ang II levels. Simulations over the ranges of ACE levels that can be achieved with ACE inhibitors predict that Ang II levels will decrease only when Ang I levels have plateaued. Comparisons of the computer simulations with our genetic experiments and with prior work of others using wide dose ranges of ACE inhibitor show a satisfactory agreement and help reconcile the apparent contradictions between the genetic and pharmacological experiments. PMID- 9403549 TI - Association between the angiotensinogen 235T-variant and essential hypertension in whites: a systematic review and methodological appraisal. AB - Recently, an allelic variant of the angiotensinogen gene (AGT 235T) has been associated with increased risk of hypertension. However, this finding has not been confirmed by all investigators. A meta-analysis was performed to examine the association between the AGT 235T-allele and hypertension in whites and to identify potential reasons for the controversial results. All relevant articles published between 1992 and 1996 were identified through multiple sources. The studies were methodologically appraised, and the frequency of the AGT 235T-allele was extracted. The 235T-allele frequency was pooled using the common odds ratio (OR) estimator by Mantel-Haenszel. Homogeneity was assessed using the Breslow-Day test. Together these studies present data on 5493 patients. The AGT 235T-allele was significantly associated with hypertension (OR: 1.20; 95% [CI]: 1.11 to 1.29; P<.0001). This association increased in studies with positive family history (OR: 1.42; 95% CI: 1.25 to 1.61, P<.0001), recruitment of cases from referral centers (OR: 1.39; 95% CI: 1.20 to 1.62, P<.0001), and more severe hypertension (OR: 1.34; 95% CI: 1.22 to 1.47, P<.0001). However, the presence of methodological problems in all studies gives rise to serious concerns regarding bias and confounding. Despite a statistically significant, albeit weak, association between the AGT 235T variant and hypertension that has been confirmed through sensitivity analysis, this finding has to be interpreted with caution, as the methodological weaknesses of the individual studies are likely to have biased the outcome of the meta-analysis. Clearly, more rigorous methods need to be applied in association studies on the genetics of human hypertension. PMID- 9403548 TI - Essential hypertension and 5' upstream core promoter region of human angiotensinogen gene. AB - The angiotensinogen (AGT) gene M235T variant is associated with essential hypertension and elevated plasma AGT concentrations, although the underlying mechanisms are unknown. Recent studies have suggested that AGCE 1 (human AGT gene core promoter element 1) located in the 5' upstream core promoter region (position -25 to -1) of the human AGT gene has an important part in the expression of AGT mRNA by binding with transcription factor AGCF 1 (human AGT gene core promoter element binding factor 1), and a mutation at -20 from adenine to cytosine (A-20C) increases the level of expression of this transcript. We therefore examined subjects with this mutation to study the association with increased plasma AGT concentrations and with essential hypertension. One hundred eighty-eight subjects receiving no antihypertensive medication were examined with regard to the correlation between A-20C and plasma AGT concentrations, and 234 subjects were studied with respect to the association between A-20C and essential hypertension. A-20C was determined by polymerase chain reaction-restriction fragment length polymorphism analysis with EcoOR 109I. Multiple regression analysis showed a weak but significant correlation between A-20C and plasma AGT concentrations (P=.047) and essential hypertension (P=.049). The results suggest that A-20C may underlie the increase in plasma AGT concentrations and be involved in the development of essential hypertension. PMID- 9403550 TI - Membrane ion transport in Bartter's syndrome: evidence for a new syndrome subtype. AB - Fifteen patients with Bartter's syndrome (hyponatremic hypochloremic hypokalemic metabolic alkalosis) were compared with 15 healthy volunteers. Red blood cell Na+/H+ and Cl-/HCO3- exchanges were enhanced in all patients with Bartter's syndrome. In calciuric normomagnesemic patients, sensitive to nonsteroidal anti inflammatory drugs (classic Bartter's syndrome), red blood cell Na+,K+,2Cl- cotransport was markedly reduced, calcium-dependent K+ permeability was moderately increased, and up to 60% of sodium permeability was represented by cAMP-activated fraction (presumably human analog of beta-isoform of Na+/H+ exchange). In noncalciuric hypomagnesemic patients insensitive to indomethacin (Gitelman's syndrome), Na+,K+,2Cl- cotransport was enhanced, Na+ permeability was increased due to calmodulin-dependent fraction, and calcium-dependent K+ permeability was markedly enhanced. A new subtype of Bartter-like syndrome ("variant Bartter's syndrome") has been described in which calciuria, hypomagnesemia, and insensitivity to nonsteroidal anti-inflammatory drugs were associated with decreased Na+,K+,2Cl- cotransport, enhanced calmodulin-activated fraction of Na+ influx, and reduced calcium-dependent K+ permeability. PMID- 9403552 TI - Regulation of the rat atrial natriuretic peptide gene after acute imposition of left ventricular pressure overload. AB - The upregulation of left ventricular (LV) atrial natriuretic peptide (ANP) mRNA is a highly conserved marker of cardiac hypertrophy. The aim of this study was to further examine the pathway leading to ANP induction during pressure overload of the heart. Systolic wall stress was imposed acutely on isovolumetrically beating rat hearts in a Langendorff apparatus (sigma-=300 x 10[3] dyne/cm2). Northern and Western blots revealed that elevated wall stress induced LV c-fos and c-jun mRNAs (3.5- and 3-fold, P<.05 after 60 minutes), c-Fos and c-Jun proteins (3.9- and 4.3 fold, P<.05 after 120 minutes), as well as ANP mRNA (2.2-fold, P<.05 after 120 minutes). ANP upregulation was prevented by inhibition of protein synthesis (cycloheximide). Electrophoresis mobility shift assays were performed to link c Fos and c-Jun (ie, components of the heterodimeric transcription factor AP-1) and ANP induction. A putative AP-1 binding site within the rat ANP promoter (nucleotides -512 to -473) bound specifically to nuclear proteins of wall stress stimulated hearts. Antibodies directed against c-Fos protein resulted in a shift of this DNA/protein complex, suggesting physical interaction between AP-1 and the ANP promoter. Myocardial transfection of promoter constructs revealed that after acute imposition of wall stress, this AP-1 site enhanced a reporter gene (8- to 10-fold compared with a minimal promoter, P<.05). Interestingly, nuclear extracts of stimulated hearts as well as pure AP-1 protein bound to a putative CRE site (nucleotides -613 to -584) as well. Like the AP-1 site, this cAMP-responsible element (CRE) site was found to enhance the transfected ANP promoter/reporter gene significantly (17.5-fold, P<.05). Mutation of either AP-1 or CRE sites did not decrease reporter gene activity, whereas mutation of both resulted in loss of inducibility. These experiments suggest that LV ANP regulation after acute wall stress includes the activation of AP-1 and/or CRE cis acting elements. However, the transient nature of c-fos and c-jun upregulation also suggests that AP-1 is not the only mediator of ANP induction in LV hypertrophy. PMID- 9403551 TI - Tissue-specific regulation of renal and cardiac atrial natriuretic factor gene expression in deoxycorticosterone acetate-salt rats. AB - Atrial natriuretic factor (ANF) is expressed in several noncardiac tissues where it may have an autocrine or paracrine function. Such function may be expected of locally synthesized ANF in the renal parenchyma. Previous investigations of the existence of ANF mRNA in the renal parenchyma have yielded conflicting results. The investigations reported here were designed to detect and measure ANF mRNA in normal rats and in rats subjected to a deoxycorticosterone acetate (DOCA)-salt treatment schedule known to strongly activate cardiac ANF gene expression. The expression of the renal ANF gene was measured using a newly developed quantitative competitive reverse transcription-polymerase chain reaction (QC-RT PCR). This method uses an internal competitor that serves as an internal standard and makes the procedure independent of measurement relative to housekeeping genes. It was found that renal ANF mRNA levels were 10(7) times lower than those found in left or right atria, but immunoreactive (ir) renal ANF concentration by specific radioimmunoassay was 10(4) times lower than that of atrial irANF levels. Reverse-phase high-performance liquid chromatography analysis revealed that more than 99% of renal irANF is processed ANF(99-126). This finding suggests that most of the irANF measured in kidney extracts likely originates from atrial sources. Left atrial ANF mRNA levels after 1 week of DOCA-salt treatment was significantly higher than that of control rats ([21.06+/-2.99] x 10(-l5) mol/microg total RNAversus [8.59 +/-1.26] x 10(-5) mol/microg total RNA, P<.05). However, renal ANF mRNA levels in DOCA-salt rats were significantly decreased compared with those of control rats ([1.64+/-0.34] x 10(-22) mol/microg total RNA versus [3.96+/-0.61]x 10(-22) mol/microg total RNA, P<.05). These results indicate that (1) renal ANF mRNA can be consistently and specifically demonstrated after reverse transcription and PCR amplification; (2) renal and cardiac ANF synthesis are regulated in a tissue-specific, opposite manner during DOCA-salt treatment; and (3) the finding that renal ANF mRNA is downregulated by DOCA-salt treatment together with previous findings suggest the need for further investigation into the role of renal ANF mRNA downregulation in the pathogenetic mechanism that leads to volume expansion and hypertension after chronic DOCA-salt treatment. PMID- 9403553 TI - Alpha-adrenergic signal transduction in renin transgenic rats. AB - The alpha1-adrenoceptor-G protein-phosphoinositide-specific phospholipase C (PLC) signal transduction pathway is assumed to play an important role in the regulation of contractile force and in the pathophysiology of myocardial hypertrophy. In the present study, the components of this pathway were investigated in left ventricles of hearts from hypertensive transgenic rats overexpressing the mouse renin gene [TG(mREN2)27] in comparison to age- and weight-matched Sprague-Dawley control rats. Contractile force was assessed in isolated electrically driven left ventricular papillary muscle strips. Alpha1 adrenoceptor density was measured by radioligand binding using [3H]prazosin, steady state levels of alpha q/11, and G protein beta-subunits by Western blotting. PLC activity was determined by a cell-free assay using exogenous phospholipid vesicles containing [3H]phosphatidylinositol (4,5)-bisphosphate as a substrate. Alpha1-adrenoceptor density was significantly increased (by 80%) in transgenic rats compared with control rats, while the positive inotropic response to the alpha1-adrenoceptor agonist phenylephrine was significantly reduced, suggesting a postreceptor defect in TG(mREN2)27. The expression of alpha q and alpha11 was verified by reverse transcription-polymerase chain reaction, and alpha q/11 steady state protein levels were shown to be similar in transgenic and control rats. Western blotting using a beta-common antibody revealed two bands at approximately 35 and 36 kD. The quantities of both were similar in TG(mREN2)27 compared with those in control rats. In contrast, PLC activity was significantly reduced (by 32%) in transgenic rats. In conclusion, our findings are consistent with a desensitization of the alpha1-adrenergic signal transduction pathway at the level of the effector. PMID- 9403554 TI - Captopril modifies gene expression in hypertrophied and failing hearts of aged spontaneously hypertensive rats. AB - The spontaneously hypertensive rat (SHR) exhibits a transition from stable compensated left ventricular (LV) hypertrophy to heart failure (HF) at a mean age of 21 months that is characterized by a decrease in alpha-myosin heavy chain (alpha-MHC) gene expression and increases in the expression of the atrial natriuretic factor (ANF), pro-alpha1(III) collagen, and transforming growth factor beta1 (TGF-beta1) genes. We tested the hypotheses that angiotensin converting enzyme inhibition (ACEI) in SHR would prevent and reverse HF associated changes in gene expression when administered prior to and after the onset of HF, respectively. We also investigated the effect of ACEI on circulating and cardiac components of the renin-angiotensin system. ACEI (captopril 2 g/L in the drinking water) was initiated at 12, 18, and 21 months of age in SHR without HF and in SHR with HF. Results were compared with those of age-matched normotensive Wistar-Kyoto (WKY) rats, and to untreated SHR with and without evidence of HF. ACEI initiated prior to failure prevented the changes in alpha MHC, ANF, pro-alpha1(III) collagen, and TGF-beta1 gene expression that are associated with the transition to HF. ACEI initiated after the onset of HF lowered levels of TGF-beta1 mRNA by 50% (P<.05) and elevated levels of alpha-MHC mRNA two- to threefold (P<.05). Circulating levels of renin and angiotensin I were elevated four- to sixfold by ACEI, but surprisingly, plasma levels of angiotensin II were not reduced. ACEI increased LV renin mRNA levels in WKY and SHR by two- to threefold but did not influence LV levels of angiotensinogen mRNA. The results suggest that the anti-HF benefits of ACEI in SHR may be mediated, at least in part, by effects on the expression of specific genes, including those encoding alpha-MHC, ANF, TGF-beta1, pro-alpha1(III) collagen, and renin angiotensin system components. PMID- 9403555 TI - Cardiac production and secretion of adrenomedullin are increased in heart failure. AB - Plasma adrenomedullin (AM) levels are reportedly increased in heart failure, but whether the cardiac production and secretion of AM is increased in heart failure remains unknown. To investigate the sites of production and secretion of AM in heart failure, we measured plasma AM levels and peptide and mRNA levels of AM in various tissues in rats with heart failure. We also examined whether the heart actually secretes AM into the circulation in patients with heart failure. We measured plasma and tissue AM levels by specific radioimmunoassay and AM mRNA by Northern blot analysis in rats with heart failure produced by aortocaval fistula. We also measured plasma AM levels in the coronary sinus and aorta in patients with left ventricular dysfunction before and after rapid right ventricular pacing. The increase in plasma AM levels in heart failure rats correlated with ventricular weight. Tissue AM levels were increased in the heart and lungs but not in the kidneys or adrenals of rats with heart failure. Similarly, tissue AM mRNA levels were also increased in the heart and lungs of heart failure rats. Plasma AM levels were higher in the coronary sinus than in the aorta in patients with left ventricular dysfunction. Rapid right ventricular pacing increased plasma atrial natriuretic peptide but not AM. These results suggest that plasma AM levels are increased in heart failure in proportion to the severity of heart failure and that cardiac production and secretion of AM is increased in heart failure rats. The lung may be another site for increased production of AM in heart failure rats. Human failing heart actually secretes AM into the circulation, and the regulation of AM secretion appears to differ from that of atrial natriuretic peptide. PMID- 9403556 TI - Reduced cardiac contractile responsiveness to isoproterenol in obese rabbits. AB - Although obesity is characterized by increased sympathetic nervous system activity, there is often a paradoxical reduction in cardiovascular end-organ response to sympathetic stimulation. Mechanisms involved in reduced sympathetic responsiveness in obesity have not been well characterized. Therefore, we determined cardiac contractile responsiveness to beta-stimulation in the obese rabbit model using both isolated heart (IH) and isolated papillary muscle (IPM) preparations. Female New Zealand White rabbits were fed control (IH: n=9; IPM: n=6) or 10% fat diets (IH: n=9; IPM: n=7) for 12 weeks. Contractile responsiveness in the IH was determined using a modified Langendorff preparation to evaluate the dose-response relationship between isoproterenol and 1) peak developed pressure/g of left ventricular wet weight and 2) maximal rate of pressure development (+dP/dt/P). Contractile responsiveness in the IPM was determined using right ventricular papillary muscles to evaluate the dose response relationship between isoproterenol and (1) peak developed tension (T)/mm2 cross-sectional area (CSA) and (2) maximal rate of tension development (dT/dt/CSA). In the IH, baseline and maximum developed pressure/g were reduced in obese rabbits by 37% and 31%, respectively (P< or =.05). In the IPM, baseline and maximum T/CSA responses were reduced in obese rabbits by 59% and 33%, respectively (P< or =.05). Potency of isoproterenol as reflected by the EC50 did not differ between lean and obese animals in either preparation. These results demonstrate that left ventricular contractility in obesity is reduced at baseline and in response to stimulation with isoproterenol and suggest that decreased responsiveness to beta-stimulation may be a factor in the obesity-related systolic dysfunction. PMID- 9403557 TI - Relation of exercise-induced myocardial ischemia to cardiac and carotid structure. AB - There is a strong relation of carotid atherosclerosis to coronary artery disease and left ventricular hypertrophy. In addition, abnormalities of carotid structure are strongly associated with abnormal left ventricular geometry and structure. However, little is known regarding the relation of exercise-induced ST depression to carotid atherosclerosis, carotid, or left ventricular structure in the absence of apparent coronary disease. The relationship of exercise ECG myocardial ischemia to the presence of carotid atherosclerosis and to carotid and left ventricular structure was assessed in 204 asymptomatic subjects free of clinical evidence of cardiovascular disease. Myocardial ischemia on the exercise ECG, defined by a chronotropically adjusted ST/HR slope of >3.47 microV/bpm, was associated with a nearly threefold greater likelihood of discrete carotid atherosclerosis (50% [6 of 12] versus 17% [29 of 192], P=.007) and with older age, male sex, higher systolic and diastolic blood pressures, greater left ventricular mass and mass index, and greater common carotid artery intimal-medial thickness and cross-sectional area index. Stepwise logistic regression analyses, including standard risk factors, revealed that only carotid artery cross sectional area index (P=.0007) and systolic blood pressure (P=.005) independently predicted an abnormal chronotropically adjusted ST/heart rate slope. Moreover, among 132 subjects with > or = 10 microV of ST-segment depression, only left ventricular mass index and carotid artery cross-sectional area index were significant predictors of the chronotropically adjusted ST/heart rate slope response. Subendocardial ischemia on the exercise ECG is strongly associated with the presence of carotid atherosclerosis and is related to systolic blood pressure, carotid artery cross-sectional area index, and left ventricular mass index, independent of age, sex, and other cardiac risk factors. These findings provide additional insights into the relation between coronary and carotid atherosclerosis and suggest that an association among ischemia and left ventricular and carotid structural abnormalities may contribute to the pathogenesis of coronary events. PMID- 9403558 TI - Mannose 6-phosphate receptor-mediated internalization and activation of prorenin by cardiac cells. AB - The binding and internalization of recombinant human renin and prorenin (2500 microU/mL) and the activation of prorenin were studied in neonatal rat cardiac myocytes and fibroblasts cultured in a chemically defined medium. Surface-bound and internalized enzymes were distinguished by the addition of mannose 6 phosphate to the medium, by incubating the cells both at 37 degrees C and 4 degrees C, and by the acid-wash method. Mannose 6-phosphate inhibited the binding of renin and prorenin to the myocyte cell surface in a dose-dependent manner. At 37 degrees C, after incubation at 4 degrees C for 2 hours, 60% to 70% of cell surface-bound renin or prorenin was internalized within 5 minutes. Intracellular prorenin was activated, but extracellular prorenin was not. The half-time of activation at 37 degrees C was 25 minutes. Ammonium chloride and monensin, which interfere with the normal trafficking and recycling of internalized receptors and ligands, inhibited the activation of prorenin. Results obtained with cardiac fibroblasts were comparable to those in the myocytes. This study is the first to show experimental evidence for the internalization and activation of prorenin in extrarenal cells by a mannose 6-phosphate receptor-dependent process. Our findings may have physiological significance in light of recent experimental data indicating that angiotensin I and II are produced at cardiac and other extrarenal tissue sites by the action of renal renin and that intracellular angiotensin II can elicit important physiological responses. PMID- 9403559 TI - Monocyte chemoattractant protein-1 expression in aortic tissues of hypertensive rats. AB - Monocyte chemoattractant protein-1 (MCP-1), a potent monocyte chemoattractant synthesized by vascular cells and monocytes, has been proposed to be an important mediator of inflammatory responses in the arterial vasculature. It was recently demonstrated that hypertension is associated with an inflammatory response in the arterial wall. To determine the effect of hypertension on arterial MCP-1 expression, we induced hypertension in Sprague-Dawley rats by infusing angiotensin II (0.75 mg x kg[-1] x d[-1] SC) for 7 days. Using Northern blot analysis, we detected a 3.6-fold increase in MCP-1 mRNA in the aortas of hypertensive rats. When we normalized blood pressure in angiotensin II-treated rats through oral administration of the nonspecific vasodilator hydralazine (15 mg x kg[-1] x d[-1]), aortic MCP-1 mRNA expression was significantly reduced. Similar results were obtained with a norepinephrine model of hypertension. Taken together, these data suggest that mechanical factors may be responsible in part for the upregulation of expression. Consistent with this interpretation, we found that cultured rat aortic vascular smooth muscle cells exposed to mechanical strain (20% peak deformation at 1 Hz) exhibited a marked increase in MCP-1 expression, suggesting the hemodynamic strain imparted onto arterial cells in hypertension is an important stimulus underlying this phenomenon. These results provide important insights into the in vivo regulation of MCP-1 and have potential implications for understanding the influence of hypertension on atherosclerosis. PMID- 9403560 TI - Increased expression of Ca2+-sensitive K+ channels in aorta of hypertensive rats. AB - Potassium efflux through Ca2+-sensitive K+ channels (K[Ca] channels) is increased in arterial smooth muscle cells from hypertensive rats, but the molecular mechanism is unknown. The goal of this study was to compare the levels of K(Ca) channel current between aortic smooth muscle cells from adult Wistar-Kyoto rats (WKY) and spontaneously hypertensive rats (SHR) and then use Western blot methods and ribonuclease protection assays to examine the expression and mRNA levels for the K(Ca) channel in these same vascular tissues. Whole-cell patch-clamp methods indicated a larger component of K(Ca) channel current, sensitive to block by iberiotoxin (100 nmol/L), in single aortic smooth muscle cells from SHR compared with WKY. Subsequent Western blot analysis using a site-specific antibody (anti alpha[913-926]) directed against the S9/S10 linker of the alpha-subunit of the K(Ca), channel revealed a 125-kD immunoreactive band in lanes loaded with either WKY or SHR aortic muscle membranes. The immunoreactive density of this band, which corresponded to the known molecular size of the alpha-subunit, was 2.2-fold greater in lanes loaded with aortic smooth muscle membranes from the hypertensive animals. However, despite this evidence for an increased expression and functional enhancement of K(Ca) channels in aortic smooth muscle membranes of SHR, ribonuclease protection assays with a 32P-labeled riboprobe targeted against the S9/S10 linker of the K(Ca) channel alpha-subunit revealed no difference in mRNA levels for the alpha-subunit between WKY and SHR aortic tissue. These findings provide initial evidence that (1) an increased expression of K(Ca) channels may be a mechanism for the enhanced K(Ca) current in aortic smooth muscle membranes of SHR, and (2) the upregulation of K(Ca) channels in arterial muscle membranes during hypertension, which is regarded as a homeostatic mechanism for buffering vascular excitability, may rely on posttranscriptional events. PMID- 9403561 TI - Pulse pressure: a predictor of long-term cardiovascular mortality in a French male population. AB - Studies on the usefulness of blood pressure as a prognostic factor in cardiovascular disease have more often involved investigations of the levels of diastolic or systolic blood pressure. However, blood pressure may be divided into two other components: steady (mean pressure) and pulsatile (pulse pressure). In this study, the relationship of pulse pressure to cardiovascular mortality was investigated in 19 083 men 40 to 69 years old who were undergoing a routine systematic health examination and were being followed up after a mean period of 19.5 years. Subjects were divided into four groups according to age (40 to 54 and 55 to 69 years) and mean arterial pressure (<107 and > or =107 mm Hg). Each group was further divided into four subgroups according to the pulse pressure level. A wide pulse pressure (evaluated according to the quartile group or as a continuous quantitative variable) was an independent and significant predictor of all-cause, total cardiovascular, and, especially, coronary mortality in all age and mean pressure groups. No significant association between pulse pressure and cerebrovascular mortality was observed. In conclusion, in a large population of men with a relatively low cardiovascular risk, a wide pulse pressure is a significant independent predictor of all-cause, cardiovascular, and, especially, coronary mortality. PMID- 9403562 TI - On-line synthesis of the human ascending aortic pressure pulse from the finger pulse. AB - Although systolic pressure in the ascending aorta (AA) can be determined accurately from the radial arterial waveform using a single generalized transfer function (TF) of the upper limb, a better on-line methods is needed for accurate noninvasive synthesis of the AA pressure contour to characterize left ventricular contractile function and ventricular-vascular coupling. AA, tonometric carotid (CA), and photoplethysmographic finger (FA) arterial pressure waveforms were recorded in 12 subjects (10 male, aged 59.1+/-10.3 years, mean+/-SD) during cardiac catheterization. The AA-FA TF was estimated using (1) a single generalized TF (GAA), (2) individualized TFs directly determined from CA-FA recordings in each patient (DAA), and (3) individualized TFs computed from CA-FA recordings in each patient with a mathematical model of the human upper limb (MAA). AA pressure waveforms were synthesized from FA recordings in real time using convolution windows derived from these TFs. Under steady state conditions, the root mean square error (RMSE) between measured and synthesized AA was lower by DAA (3.3+/-1.3 mm Hg) and MAA (3.9+/-1.2 mmHg) than by GAA (4.8+/-2.0 mm Hg, P<.05). During dynamic load alteration induced by the Valsalva maneuver, however, the MAA method performed better (5.4+/-2.8 mm Hg) than both the GAA (5.8+/-3.3 mm Hg, P<.05) and DAA (6.5+/-2.7 mm Hg, P<.01) methods. The beat-to-beat AA contour can be accurately and noninvasively synthesized on-line using individualized TFs. During dynamic load alteration, individualized TFs derived with an upper limb arterial model provide greater accuracy. PMID- 9403563 TI - Increased stiffness of radial artery wall material in end-stage renal disease. AB - The incremental elastic modulus (Einc), which is the slope of the relationship between stress and strain of arteries, is a marker of vascular wall material stiffness. Isobaric Einc is reduced at the site of the radial artery in patients with essential hypertension and increased at the site of the common carotid artery in subjects with end-stage renal disease (ESRD). Whether the changes in Einc are influenced by the topography of the vessels, the composition of the arterial wall, and/or by the presence of ESRD is largely ignored. Radial artery Einc was measured in 19 patients with ESRD and compared with the Einc of 89 subjects with essential hypertension and 20 normotensive control subjects. Transcutaneous measurements of radial artery internal diameter and wall thickness (echo-tracking device) and digital pulse pressure (Finapres) were allowed to calculate Einc under operational (ie, at the mean arterial pressure of each group) and isobaric (100 mm Hg) conditions, as well as for a given wall stress. Internal diameter and pulsatile changes in diameter were identical in the three groups. Wall thickness and mean blood pressure were significantly elevated in subjects with hypertension but not in ESRD patients. Circumferential wall stress was identical in the three groups. For the same operational wall stress, and therefore at the operational mean arterial pressure of each group, Einc (kPa x 10[3]) was increased in patients with ESRD (5.53+/-4.0 versus 3.3+/-2.4 in control subjects; P<.05) and normal in subjects with essential hypertension (3.87+/-4.0). Under isobaric conditions, Einc was also significantly lower in subjects with hypertension and elevated in patients with ESRD. Thus, at the site of a medium-sized muscular artery constantly devoid of atherosclerosis, the stiffness of wall material is increased in patients with ESRD. The demonstrated alterations of the arterial wall are independent of the level of blood pressure and tensile stress and should be related to the status ESRD. PMID- 9403564 TI - Perivascular sensory nerve Ca2+ receptor and Ca2+-induced relaxation of isolated arteries. AB - The present study tested two hypotheses: (1) that a receptor for extracellular Ca2+ (Ca2+ receptor [CaR]) is located in the perivascular sensory nerve system and (2) that activation of this receptor by physiological concentrations of extracellular Ca2+ results in the release of vasodilator substance that mediates Ca2+-induced relaxation. Reverse transcription-polymerase chain reaction using primers derived from rat kidney CaR cDNA sequence showed that mRNA encoding a CaR is present in dorsal root ganglia but not the mesenteric resistance artery. Western blot analysis using monoclonal anti-CaR showed that a 140-kD protein that comigrates with the parathyroid CaR is present in both the dorsal root ganglia and intact mesenteric resistance artery. Immunocytochemical analysis of whole mount preparations of mesenteric resistance arteries showed that the anti-CaR stained perivascular nerves restricted to the adventitial layer. Biophysical analysis of mesenteric resistance arteries showed that cumulatively raising Ca2+ from 1 to 1.25 mol/L and above relaxes precontracted arteries with an ED50 value of 2.47+/-0.17 mmol/L (n=12). The relaxation is endothelium independent and is unaffected by blockade of nitric oxide synthase but is completely antagonized by acute and subacute phenolic destruction of perivascular nerves. A bioassay showed further that superfusion of Ca2+ across the adventitial surface of resistance arteries releases a diffusible vasodilator substance. Pharmacological analysis indicates that the relaxing substance is not a common sensory nerve peptide transmitter but is a phospholipase A2/cytochrome P450-derived hyperpolarizing factor that we have classified as nerve-derived hyperpolarizing factor. These data demonstrate that a CaR is expressed in the perivascular nerve network, show that raising Ca2+ from 1 to 1.25 mol/L and above causes nerve-dependent relaxation of resistance arteries, and suggest that activation of the CaR induces the release of a diffusible hyperpolarizing vasodilator. We propose that this system could serve as a molecular link between whole-animal Ca2+ balance and arterial tone. PMID- 9403565 TI - Growth factors mediate intracellular signaling in vascular smooth muscle cells through protein kinase C-linked pathways. AB - Intracellular Ca2+ and pH are potent modulators of growth factor-induced mitogenesis and contraction. This study examined platelet-derived growth factor (PDGF-BB) and insulin-like growth factor (IGF-1)-mediated signal transduction in primary cultured unpassaged vascular smooth muscle cells (VSMC) from mesenteric arteries of Sprague-Dawley rats. Intracellular free Ca2+ concentration ([Ca2+]i) and intracellular pH (pHi) were measured by fluorescence digital imaging using fura-2 AM and 2'7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein, respectively. Characteristics of [Ca2+]i transients were determined by pre-exposing cells to Ca2+-free buffer, and involvement of the Na+/Ca2+ exchanger was assessed by withdrawal of extracellular Na+ and by exposure to dimethylbenzamil (Na+/Ca2+ exchange blocker). To determine whether pHi responses were mediated via the Na+/H+ exchanger, cells were preincubated with 10(-5) mol/L 5-(N-ethyl-N isopropyl)amiloride (a selective Na+/H+ exchange blocker). The role of protein kinase C (PKC) and tyrosine kinases in growth factor signaling was assessed by pre-exposing cells to calphostin C and chelerythrine chloride (selective PKC inhibitors; 10(-5) mol/L) and tyrphostin A23 (a selective tyrosine kinase inhibitor; 10(-5) mol/L). PDGF-BB and IGF-1 (1 to 10 ng/mL) increased [Ca2+]i and pHi in a dose-dependent manner. At concentrations greater than 1 ng/mL both growth factors induced a biphasic [Ca2+]i response with an initial transient peak followed by a sustained elevation. At 5 ng/mL PDGF-BB and IGF-1 significantly increased [Ca2+]i from 95+/-3 nmol/L to 328+/-28 and 251+/-18 nmol/L, respectively. Ca2+ withdrawal abolished the second phase of [Ca2+]i elevation. Agonist-induced [Ca2+]i responses were similarly altered by Na+ withdrawal, by Na+/ Ca2+ exchange blockade, and by PKC inhibition; latency, the period from stimulus application to the first [Ca2+]i peak, was increased, the initial [Ca2+]i peak was attenuated, and the sustained phase was prolonged. PDGF-BB and IGF-1 (10 ng/mL) significantly increased pHi from 6.89+/-0.04 nmol/L to 7.11+/ 0.01 and 7.09+/-0.02 nmol/L, respectively. EIPA and calphostin C completely inhibited agonist-elicited alkalinization. Tyrphostin A-23 abolished second messenger responses to PDGF-BB and IGF-1, whose receptors have tyrosine kinase activity. In conclusion, PDGF-BB and IGF-1 elicit significant [Ca2+]i and pHi responses in VSMC. The underlying pathways that mediate these responses are partially dependent on Na+/ Ca2+ transporters and the Na+/H+ exchanger, both of which are linked to PKC activation. PMID- 9403566 TI - Role of angiotensin II in the regulation of a novel vascular modulator, hepatocyte growth factor (HGF), in experimental hypertensive rats. AB - Hepatocyte growth factor (HGF) is a mesenchyme-derived pleiotropic factor that regulates cell growth, cell motility, and morphogenesis of various types of cells, and is thus considered a humoral mediator of epithelial-mesenchymal interactions responsible for morphogenic tissue interactions. We have previously reported that HGF is a novel member of endothelium-specific growth factors whose serum concentration is positively associated with blood pressure in humans. Therefore, we speculated that serum HGF secretion might be elevated in response to high blood pressure as a counter-system against endothelial dysfunction. However, it is difficult to elucidate the role of circulating and tissue HGFs in human hypertension. To address this issue, we measured circulating and tissue HGF concentrations in spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY) at different ages. Serum HGF concentration in SHR was significantly higher than that in WKY at 6, 15, and 25 weeks of age (P<.01). Serum HGF concentration was also significantly positively correlated with blood pressure in SHR (P<.02, r=.455). In contrast, tissue HGF concentrations in heart, aorta, and kidney were significantly decreased in SHR as compared with WKY at 25 weeks of age, when these organs showed hypertrophic changes induced by hypertension (P<.01). Cardiac HGF mRNA was also decreased in SHR as compared with WKY at 25 weeks of age. Moreover, cardiac HGF concentration showed a significant negative correlation with left ventricular (LV) weight (P<.01), whereas serum HGF concentration showed a significant positive correlation with LV weight (P<.05). Interestingly, concentrations of cardiac and vascular angiotensin II, a suppressor of HGF, were increased in SHR as compared with WKY at 25 weeks of age (P<.01). Therefore, we examined the effects of angiotensin blockade on circulating and tissue HGF concentrations, to study the role of angiotensin II in HGF regulation. Administration of an angiotensin-converting enzyme inhibitor (enalapril) and angiotensin II type 1 receptor antagonists (losartan and HR 720) for 6 weeks resulted in a significant increase in cardiac HGF concentration, accompanied by increased cardiac HGF mRNA, and a significant decrease in serum HGF concentration, accompanied by lowered blood pressure and reduced LV weight (P<.01). Here, we demonstrated increased circulating HGF and decreased vascular, cardiac, and renal HGF in SHR as compared with WKY at the maintenance stage of hypertension. Decreased tissue HGF in target organs of hypertension may be due to increased tissue angiotensin II. These results suggest that decreased local HGF production may have an important role in the cardiovascular remodeling of target organs in hypertension, since HGF prevented endothelial injury and promoted angiogenesis. Blockade of angiotensin augmented local decreased cardiovascular HGF in hypertension, potentially resulting in the improvement of endothelial dysfunction. PMID- 9403567 TI - Cellular aspects of vascular remodeling in hypertension revealed by confocal microscopy. AB - Cellular aspects of remodeling in intact arteries have not been fully investigated, mainly due to the lack of an appropriate methodology that allows for simple measurements. The aim of this study was to develop a method based on laser scanning confocal microscopy (LSCM), compare it with previous methodology, and apply it to the study of remodeling in hypertension. The morphology of mesenteric resistance arteries from stroke-prone spontaneously hypertensive rats (SHRSP) and Wistar-Kyoto rats (WKY) was determined with wire myography on one segment with a standardized diameter setting (0.9[d100]) and with perfusion myography on a second segment from the same artery at the calculated equivalent pressure. The second segments were stained with the nuclear dye Hoechst 33342 (live tissue) or propidium iodide (fixed tissue) and measured with LSCM and MetaMorph software. Compared with wire myography, perfusion myography showed similar differences from those previously reported. Compared with LSCM, perfusion myography showed a similar lumen but significantly smaller wall thickness in both live and fixed tissue, probably due to measurement underestimation. In the study with LSCM, arteries from SHRSP compared with those from WKY showed (1) reduced lumen, (2) altered cell density that was significantly increased in the adventitia, decreased in the media, and unchanged in the intima, (3) significantly increased medial volume, (4) significantly smaller endothelial cell nuclei, and (5) adventitial-like cells in the media. We conclude that (1) LSCM is a reliable and straightforward method to study morphology in intact vessels, (2) it provides new information on the cellular changes in remodeling, (3) adventitia might play an active role in the process of remodeling in hypertension, and (4) endothelium "remodels" in hypertension. PMID- 9403569 TI - AT1 receptor antagonist treatment caused persistent arterial functional changes in young spontaneously hypertensive rats. AB - The effects of chronic treatment with an AT1 receptor antagonist (L-158,809) on hypertension development and cardiovascular changes were studied in spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY). L-158,809 treatment (0.6 mg/kg PO) was initiated at 3 weeks of age and lasted 12 weeks, to 15 weeks of age. The treatment prevented hypertension development in the SHR (systolic blood pressure, BP, of 136+/-1 mm Hg compared with 198+/-3 mm Hg in control SHR), and lowered the BP of WKY (99+/-2 vs 128+/-1 mm Hg in control WKY). Treatment significantly reduced the heart weight in SHR and WKY. Ten weeks after treatment withdrawal (25 weeks of age), BP had increased in SHR and WKY to 172+/-8 and 117+/-3 mm Hg, respectively. Body weight and kidney weight were not affected by the treatment. Mesenteric arteries from treated SHR were less responsive than control SHR arteries to periarterial nerve stimulations at transmural pressures higher than 80 mm Hg (15 and 25 weeks). Control WKY arteries were less responsive than control SHR arteries at almost all transmural pressures tested (15 weeks) and to pressures greater than 80 mm Hg (25 weeks). Pretreatment of arteries with 10(-8) mol/L angiotensin II enhanced their response to nerve stimulation in vessels from control SHR and WKY (25 weeks) but not from treatment-withdrawn SHR and WKY. Treatment did not alter arterial reactivity in response to norepinephrine. Alteration in arterial structure due to L-158,809 treatment was found only when measured at a transmural pressure of 100 mm Hg. In conclusion, L 158,809 was effective in preventing hypertension during the treatment period, in reducing hypertension severity during the withdrawal period, and in persistently decreasing the reactivity of the arteries. PMID- 9403568 TI - Role of nitric oxide in the regulation of the mechanical properties of peripheral conduit arteries in humans. AB - Whether nitric oxide (NO) contributes to the regulation of the mechanical properties of large arteries in humans is not known. We measured the effect of local administration of the inhibitor of NO synthesis N(G)-monomethyl-L-arginine (L-NMMA; 1 and 4 micromol x L[-1] x min[-1] for 5 minutes) and acetylcholine (3 and 30 nmol x L[-1] x min[-1] for 3 minutes) on radial artery diameter and wall thickness in 11 healthy volunteers using an echo-tracking system coupled to a measurement of radial blood flow (Doppler) and arterial pressure. At the highest dose, L-NMMA reduced radial blood flow but surprisingly decreased incremental elastic modulus (from 1.36+/-0.22 to 1.00+/-0.22 kPa x 10[3]; P<.05) and increased arterial compliance (from 3.20+/-0.46 to 4.07+/-0.45 m2 x kPa x 10(-8), P<.05), without affecting radial artery internal diameter, wall thickness or midwall stress, thus reflecting a decrease in vascular tone. Acetylcholine decreased incremental elastic modulus (from 1.27+/-0.08 to 0.88+/-0.07 kPa x 10[3]; P<.05) and increased arterial diameter, radial blood flow, and compliance (from 2.82+/-0.16 to 5.30+/-0.62m2 x kPa x 10[-8]; P<.05). These results demonstrate in vivo that NO is involved in the regulation of the mechanical properties of large arteries in humans. However, the effects of L-NMMA, ie, a decrease in arterial wall rigidity and an increase in arterial compliance, which occur in the absence of any changes in blood pressure or arterial geometry, suggest that inhibition of NO synthesis is associated in humans with a paradoxical isometric smooth muscle relaxation. This effect could be due to the development of compensatory vasodilating mechanisms after NO synthesis inhibition. PMID- 9403570 TI - Regional renal nitric oxide release in stroke-prone spontaneously hypertensive rats. AB - Diminished nitric oxide (NO) production has been implicated in the pathogenesis of salt-sensitive hypertension. We questioned whether such a defect is responsible for the malignant hypertension and nephrosclerosis in stroke-prone spontaneously hypertensive rats (SHRSP) fed a high-salt/stroke-prone diet (S) versus a regular diet (R). NO release from 30-minute incubates of cortex and outer and inner medulla were studied in SHRSP at 10, 12, and 16 weeks of age on the S diet versus R diet. SHRSP-S (n=16) exhibited a marked age-dependent increase in NO release, especially in the cortex. Increases were only modest in SHRSP-R (n=21). At 16 weeks, cortical NO was 93+/-25 versus 6+/-1 pmol/mg tissue in SHRSP-S versus SHRSP-R (P<.001). Immunohistochemical staining increased mostly for neuronal, slightly for endothelial, and negligibly for inducible isoforms of NO synthase and was predominantly in the cortex of SHRSP-S versus SHRSP-R. Despite similar hypertension in SHRSP-S versus SHRSP-R (mean arterial pressure, 174+/-7 versus 177+/-2 mm Hg), malignant nephrosclerosis was seen only in SHRSP S, affecting 22+/-6% of glomeruli and 23+/-4 vessels per 100 glomeruli by 16 weeks. N omega-nitro-L-arginine (15 mg/kg per day) in SHRSP-S (n=6) abrogated the increase in cortical NO but further augmented the hypertension and accelerated lesion development. Wistar-Kyoto rats at 16 weeks on the R diet (n=8) had NO levels similar to those of SHRSP-R, showed increased cortical NO to only 28+/-10 pmol/mg on the S diet (n=9) (P<.05 versus SHRSP-S), but remained normotensive and lesion-free. We conclude that hypertension and lesion development in SHRSP are not due to deficient renal NO. Accelerated onset of malignant nephrosclerosis by NO synthase inhibition suggests that NO is protective in these animals, mitigating the effects of hypertension and S diet on renal pathology. PMID- 9403571 TI - Lead-induced hypertension: interplay of nitric oxide and reactive oxygen species. AB - An elevation of mean blood pressure was found in rats treated with low lead (0.01% lead acetate) for 3 months, as contrasted to paired Sprague-Dawley control rats. In these rats, measurement of plasma and urine endothelins-1 and -3 revealed that plasma concentration and urinary excretion of endothelin-3 increased significantly after 3 months (plasma: lead group, 31.8+/-2.2, versus controls, 23.0+1.7 pg/mL, P<.001; urinary excretion: lead group, 46.6+11.7, versus controls, 35.6+6.7 pg/24 h, P<.05), whereas endothelin-1 was unaffected. Plasma and urinary nitric oxide (NO) and cyclic GMP concentrations were not significantly changed. However, assay of plasma and kidney cortex malondialdehyde by high-pressure liquid chromatography, as a measure of reactive oxygen species, was elevated in lead-treated rats compared with that in control rats (plasma: lead group, 4.74+1.27, versus controls, 2.14+.49 micromol/L, P<.001; kidney cortex: lead group, 28.75+3.46, versus controls, 16.38+2.37 nmol/g wet weight, P<.001). There was increased NO synthase activity in lead-treated rat brain cortex and cerebellum. In lead-treated rat kidney cortex, the endothelial constitutive NO synthase protein mass was unaffected, whereas the inducible NO synthase protein mass was increased. These data suggest a balance between increased NO synthesis and degradation (by reactive oxygen species) in lead treated rats, which results in normal levels of NO. Thus, the hypertension may be related to an increase in the pressure substances, endothelin-3 and reactive oxygen species, rather than to an absolute decrease in nitric NO. PMID- 9403572 TI - Hypothalamic hypertensive factor: an inhibitor of nitric oxide synthase activity. AB - Human and rat plasma and rat hypothalamus contain a cytochemically detectable substance, the concentration of which rises with an increase in salt intake. The plasma concentration of this material is also raised in essential hypertension and in the spontaneously hypertensive rat (SHR), the Milan hypertensive rat, and the reduced renal mass (RRM) hypertensive rat. In the normal rat, the greatest concentration is found in the hypothalamus of the SHR and the RRM hypertensive rat. The physicochemical characteristics of this cytochemically detectable hypothalamic hypertensive factor (HHF), including chromatographic behavior and molecular weight range, suggest that it may share features common to a substituted guanidine that is present in established nitric oxide synthase (NOS) inhibitors. It was therefore decided to determine the effect on NOS activity of the HHF obtained from mature SHR. The ability of HHF to inhibit NOS activity was studied on (1) NOS extracted from bovine aorta, rat brain, and human platelets by measuring the conversion of radiolabeled L-arginine to L-citrulline and (2) rat liver NOS measured indirectly with a cytochemical technique based on the stimulation of soluble guanylate cyclase activity in hepatocytes by NO. HHF showed a biphasic inhibitory action on platelet NOS activity that was greater with HHF obtained from SHR than from Wistar-Kyoto rats. HHF also had a biphasic inhibitory effect on hepatocyte NOS activity that was more potent when obtained from SHR. It is proposed that the increase in HHF, a novel form of NOS inhibitor that is elevated in SHR, may be involved in the rise in arterial pressure. PMID- 9403573 TI - Cardiovascular effects of nitric oxide and N-methyl-D-aspartate receptors in the nucleus tractus solitarii of rats. AB - Nitric oxide (NO) is an endogenously synthesized effector molecule that acts as a neurotransmitter with novel properties in both the central and peripheral nervous systems. We previously reported that NO was involved in central cardiovascular regulation and modulated the baroreflex in the nucleus tractus solitarii (NTS) of rats. The aim of the present study was to determine whether NO and excitatory amino acids reciprocally release each other in the NTS. In normotensive Sprague Dawley rats, intra-NTS microinjection of L-arginine (1 to 100 nmol/60 nL) produced a dose-dependent decrease in blood pressure and heart rate. Microinjection of excitatory amino acids L-glutamate and NMDA also produced depressor and bradycardic effects. These effects of L-glutamate or NMDA were blocked by prior administration of NO synthase inhibitor N(G)-methyl-L-arginine or N(G)-nitro-L-arginine methyl ester. Similarly, prior administration of N methyl-D-aspartate (NMDA) receptor antagonist MK-801 and non-NMDA receptor antagonist 6,7-dinitroquinoxaline-2,3-dione significantly attenuated the depressor and bradycardic effect of L-arginine. These results demonstrated a reciprocal attenuation of NO synthase inhibitor and NMDA receptor antagonist on NMDA and L-arginine responses, respectively, in the NTS and suggest that NO and NMDA receptors may interact in central cardiovascular regulation. PMID- 9403574 TI - Autonomic nervous system dysfunction in elderly hypertensive patients with abnormal diurnal blood pressure variation: relation to silent cerebrovascular disease. AB - To investigate the relationships among diurnal blood pressure (BP) variations and autonomic nervous system dysfunction, we assessed heart rate variability (HRV) using power spectral analysis of the 24-hour RR interval in 51 asymptomatic elderly hypertensive patients with various patterns of nocturnal BP fall. The extreme-dippers with marked nocturnal BP fall (n=16) had lower asleep low frequency power (LF)/high-frequency power (HF) ratios (a relative index of sympathetic nervous system activity), while the nondippers without nocturnal BP fall (n=18) had lower awake LF/HF ratios and asleep/awake ratio for HF (an index of parasympathetic nervous activity), when compared with dippers with appropriate nocturnal BP fall (n=17). The incidence of multiple lacunar infarction detected by brain magnetic resonance imaging was 56% in the extreme-dippers and 38% in the nondippers, and both were markedly higher than that (6.3%) in the dippers (both P<.01). There was no significant relationship between the BP level and any HRV parameter for either the daytime or nighttime period. The asleep/awake ratio for systolic BP was significantly correlated with the asleep/awake ratio for HF (r= .363, P<.01) and with the asleep/awake ratio for the LF/HF ratio (r=.540, P<.001), regardless of whether multiple lacunar infarction was present. In conclusion, the autonomic nervous system activity is not related to high BP level per se, rather its diurnal variation is more important as a determinant of the diurnal BP patterns, regardless of the presence or absence of cerebrovascular disease. PMID- 9403575 TI - Relationship between blood pressure and body mass index in lean populations. AB - Associations between body mass index (BMI) and blood pressure (BP) have been consistently observed, but remain poorly understood. One unresolved question is whether there is a linear relationship across the entire BMI range. We investigated this question among 11,235 adult men and women from seven low-BMI populations in Africa and the Caribbean. We used kernel smoothing and multivariate linear and spline regression modeling to examine gender differences in the relationship and to test for a threshold. Age-adjusted slopes of BP on BMI were uniformly higher in men than women, with pooled slope ratios of 2.00 and 2.20 for systolic and diastolic BPs, respectively. Men displayed no evidence of age modification or nonlinearity in the relationship, and the age-adjusted slope of systolic BP on BMI was 0.90 (95% confidence interval [CI], 0.76 to 1.04). Women demonstrated both age modification and nonlinearity. For both younger (<45 years) and older (45+ years) women, the optimal change point for a single threshold model was found to be 21 kg/m2. Slopes of systolic BP on BMI above this threshold were positive and significant: 0.68 (95% CI, 0.54 to 0.81) and 0.53 (95% CI, 0.29 to 0.76) for younger and older women, respectively. Slopes below the threshold were essentially zero for both groups of women, and the difference between the slopes above and below the threshold was significant for younger women (P=.019). In summary, we observed a threshold at 21 kg/m2 in the relationship between BMI and BP for women but not for men. This contributes to the effort to identify the mechanisms that underlie this relationship and how they differ by gender. PMID- 9403576 TI - Sex differences in the abundance of endothelial nitric oxide in a model of genetic hypertension. AB - A deficiency of nitric oxide may be responsible for the increased vascular resistance associated with human essential hypertension and that seen in animal models of hypertension. Premenopausal females are relatively protected from hypertension and cardiovascular complications. Levels of superoxide can influence the availability of nitric oxide. We hypothesize that there are differences in nitric oxide availability between stroke-prone spontaneously hypertensive rats (SHRSP) and Wistar-Kyoto rats (WKY) and that superoxide may be responsible for at least some of these differences. We studied vascular reactivity in endothelium intact aortic rings from WKY and SHRSP. We measured nitric oxide synthase activity in endothelial cells removed from aortas and also measured circulating nitrite/nitrate levels. We found the response to N(G)-nitro-L-arginine methyl ester to be significantly greater in WKY compared with SHRSP (95% CI: 20 to 174; P=.015) and in females compared with males in WKY (95% CI: 143 to 333; P=.00004) and SHRSP (95% CI: 70 to 224; P=.0006). Endothelial nitric oxide synthase activity was significantly greater in SHRSP compared with WKY (95% CI: 2.3 to 17.6; P=.016). The EC50 for relaxation to carbachol was significantly greater in male rats compared with female rats (95% CI: -1.1 to -0.2; P=.003) within the SHRSP strain. The maximum relaxation to carbachol was significantly attenuated in stroke prone spontaneously hypertensive compared with Wistar-Kyoto rats (95% CI: 1.7 to 14.4; P=.015). Diethyldithiocarbamate had a significantly greater effect on the stroke prone spontaneously hypertensive rats' carbachol response than that of Wistar-Kyoto rats (95% CI: 14.3 to 47.0; P=.0008). We conclude that superoxide may be responsible for strain differences in vascular reactivity, whereas nitric oxide availability may be responsible for sex differences independently of endothelial nitric oxide synthase activity and superoxide. PMID- 9403577 TI - Maintenance of maternal diet-induced hypertension in the rat is dependent on glucocorticoids. AB - Recent epidemiological evidence suggests that adult cardiovascular risk is determined by birth weight and factors that influence birth weight, such as maternal nutrition. Data from animal models suggest that an interaction between nutrition and glucocorticoid hormones "programs" increased risk of adult hypertension. Increased fetal exposure to maternal glucocorticoids that is proposed to occur from a reduction in the placental barrier to maternal glucocorticoid, 11beta-hydroxysteroid dehydrogenase, is suggested to program hypertension in the resultant offspring from both glucocorticoid-treated and maternally protein-restricted rats. The extent to which postnatal glucocorticoid stimulation may influence the progression of hypertension in the offspring from protein-restricted rat dams was assessed in 6-week-old male Wistar rats, prenatally exposed to either an 18% casein (control) or 9% casein (low protein) diet. Rats from each dietary group were sham operated, adrenalectomized or adrenalectomized, and treated with 20 mg corticosterone/kg body weight per day. Before surgery, systolic blood pressure was significantly higher in the low protein-exposed rats compared with controls (165+/-3.8 versus 142+/-3.3 mm Hg, P<.0001). Adrenalectomy of the low protein-exposed animals significantly reduced the blood pressure to control levels, while corticosterone replacement restored the hypertensive state. No effect of adrenalectomy on blood pressure was observed in 18% casein controls. In both dietary groups adrenalectomy decreased brain, but not hepatic, glucocorticoid-sensitive enzyme activities and corticosterone treatment elevated activities of all enzymes. The data suggest that maternal diet induced hypertension is dependent on an intact adrenal gland postnatally and that glucocorticoids are key trophic agents in maintaining the high blood pressure. PMID- 9403578 TI - Diagnosing gestational hypertension and preeclampsia with the 24-hour mean of blood pressure. AB - The use of ambulatory blood pressure monitoring has provided a method of blood pressure assessment that may compensate for some of the limitations of isolated measurements. Here we aim to examine prospectively the effectiveness of the commonly used 24-hour mean as a potential screening test for the identification of gestational hypertension and preeclampsia. We analyzed 503 blood pressure series from 71 healthy pregnant women and 256 series from 42 women who developed gestational hypertension or preeclampsia. Forty-eight-hour blood pressure monitoring was done once every 4 weeks after the first obstetric consultation. Sensitivity and specificity of the 24-hour mean of blood pressure were computed for each trimester of pregnancy by comparing distributions of values obtained for healthy and complicated pregnancies, without assuming an a priori threshold for diagnosing gestational hypertension on the basis of mean blood pressure. Sensitivity ranges from 31.8% for diastolic blood pressure in the second trimester to 84.1% for systolic blood pressure in the third trimester. However, specificity is as low as 6.9% for diastolic blood pressure in the first trimester. The positive predictive value does not reach 55% for any variable in any trimester. The higher relative risk was consistently obtained for systolic blood pressure (4.9 in the third trimester). Despite the highly statistically significant differences in blood pressure found between healthy and complicated pregnancies in all trimesters, the daily mean of blood pressure does not provide a proper and stable individualized test for diagnosing hypertensive complications in pregnancy. Other indexes obtained from the blood pressure series have been shown, however, to identify early in pregnancy those women who subsequently will develop gestational hypertension or preeclampsia, rendering ambulatory blood pressure monitoring a useful, but still costly, technique in pregnancy. PMID- 9403579 TI - Estrogen supplementation decreases norepinephrine-induced vasoconstriction and total body norepinephrine spillover in perimenopausal women. AB - Estrogens are reported to provide protection against the development of cardiovascular disease in women, but the mechanisms underlying these effects are not well defined. We hypothesized that estrogen might reduce neural cardiovascular tone. We therefore studied responses to exogenous norepinephrine and norepinephrine spillover in 12 perimenopausal women randomized to 8 weeks of estrogen supplementation (estradiol valerate, 2 mg daily, n=7) or placebo (n=5). Forearm blood flow was measured by venous occlusion plethysmography, and vasoactive agents were infused through a brachial artery cannula in doses that did not influence blood pressure or heart rate. Total body and forearm norepinephrine spillover were measured by radiotracer methodology. Forearm vasoconstrictor responses to norepinephrine (25, 50, and 100 ng/min) were attenuated after estrogen supplementation (P=.002). Vasoconstrictor responses to angiotensin II (8, 16, and 32 ng/min) were unchanged postestrogen. There was a significant reduction in total body spillover of norepinephrine after estrogen supplementation (pre-estrogen, 700+/-152; postestrogen, 439+/-150 ng/min; P<.05), but there was no change after placebo. Total body clearance and forearm spillover of norepinephrine were unchanged by either estrogen or placebo. Estrogen supplementation also significantly decreased both systolic and diastolic blood pressures. Therefore, estrogen supplementation in perimenopausal women selectively attenuates vasoconstrictor responses to norepinephrine and reduces total body norepinephrine spillover, which is an index of sympathetic neural activity. PMID- 9403580 TI - Effect of carbidopa on the excretion of sodium, dopamine, and ouabain-like substance in the rat. AB - The effect of carbidopa, a competitive inhibitor of dopamine synthesis on the excretion of sodium, dopamine (DA), and endogenous sodium transport inhibitor (ESTI), was examined in rats on normal and high salt diets for 7 days. ESTI activity was measured (1) as ouabain-like substance (OLS) by radioimmunoassay using an antibody raised against ouabain, (2) by inhibition of purified Na+,K+ ATPase activity (ATPI), and (3) as digoxin-like substance (DLS) using a commercial digoxin assay. The OLS immunoassay was validated against rubidium transport studies. High salt diet caused an increase in OLS and ATPI but not DLS. Sodium excretion in rats on normal sodium intake was not affected by carbidopa treatment but was significantly lower in the high sodium diet group during the first 5 days of the study. In the low salt group, carbidopa treatment caused significant increases in OLS. We conclude that ESTI (measured as OLS) and DA are important in the natriuresis of salt loading. PMID- 9403581 TI - Improvement of insulin sensitivity by short-term exercise training in hypertensive African American women. AB - African American women have a high prevalence of insulin resistance, non-insulin dependent diabetes mellitus, obesity, and hypertension that may be linked to low levels of physical activity. We sought to determine whether 7 days of aerobic exercise improved glucose and insulin metabolism in 12 obese (body fat >35%), hypertensive (systolic blood pressure > or =140 and/or diastolic blood pressure > or =90 mmHg) African American women (mean age 51+/-8 years). Insulin-assisted frequently-sampled intravenous glucose tolerance tests were performed at baseline and 14 to 18 hours after the 7th exercise session. There was no significant change in maximal oxygen consumption, body composition, or body weight after the 7 days of aerobic exercise. The insulin sensitivity index increased (2.68+/-0.45 x 10[-5] to 4.23+/-0.10 x 10[-5] [min(-1)/pmol/L], P=.02). Fasting (73+/-9 to 50+/-9 pmol/L, P=.02) and glucose-stimulated (332+/-58 to 261+/-45 pmol/L, P=.05) plasma insulin levels decreased. Additional measures related to the insulin resistance syndrome also changed with the 7 days of exercise: basal plasma norepinephrine concentrations were reduced (2.46+/-0.27 to 1.81+/-0.27 nmol/L, P=.02) and sodium excretion rate increased from 100+/-13 to 137+/-7 mmol/d (P=.03); however, there was no change in potassium excretion or 24-hour ambulatory blood pressure. We conclude that a short-term aerobic exercise program improves insulin sensitivity in African American hypertensive women independent of changes in fitness levels, body composition, or body weight. The present study indicates that short-term exercise can improve insulin resistance in hypertensive, obese, sedentary African American women and confirms previous reports that a portion of the exercise-induced improvements in glucose and insulin metabolism may be the result of recent exercise. PMID- 9403582 TI - Relation of fasting insulin to blood pressure and lipids in adolescents and parents. AB - This study was intended to clarify the relation between fasting insulin, lipids, and blood pressure in adolescents before the onset of hypertension and to examine the association of these data with similar data obtained in their parents. The participants in this study were 183 adolescents 14 to 18 years old (96 girls) completing a 4-year intervention trial and their parents (164 mothers, 122 fathers). Blood pressure was measured twice on the right arm in a seated position using a random-zero sphygmomanometer. Fasting blood samples were obtained for lipid and insulin analyses. Fasting insulin was significantly correlated with systolic blood pressure in the adolescents and also in the parents before and after adjustment for body mass index. Fasting insulin was correlated significantly with levels of cholesterol, triglycerides, and HDL and LDL cholesterol in the adolescents. It was correlated only with triglycerides and HDL cholesterol in mothers and fathers. After adjustment for body mass index, the correlations between fasting insulin and lipids in the children were not significant. A significant relation was shown between children's systolic blood pressure and mothers' fasting insulin and systolic blood pressure. Significant correlations were found between the children's and fathers' triglycerides and HDL cholesterol, whereas significant correlations were found for fasting insulin and all lipids between mothers and children, and these remained significant after adjustment for body mass index. These results show (1) a significant relation between fasting insulin and both lipids and systolic blood pressure in adolescents and (2) a significant relation for these factors between adolescents and their parents. Although weight appears to play an important role in this relation during adolescence, genetic and environmental factors other than those mediated via weight may control insulin metabolism within families. The data support a role for studies during early biological development to address these issues. PMID- 9403583 TI - Antihypertensive agent moxonidine enhances muscle glucose transport in insulin resistant rats. AB - The sympatholytic antihypertensive agent moxonidine, a centrally acting selective I1-imidazoline receptor modulator (putative agonist), may be beneficial in hypertensive patients with insulin resistance. In the present study, the effects of chronic in vivo moxonidine treatment of obese Zucker rats--a model of severe glucose intolerance, hyperinsulinemia and insulin resistance, and dyslipidemia- on whole-body glucose tolerance, plasma lipids, and insulin-stimulated skeletal muscle glucose transport activity (2-deoxyglucose uptake) were investigated. Moxonidine was administered by gavage for 21 consecutive days at 2, 6, or 10 mg/kg body weight. Body weights in control and moxonidine-treated groups were matched, except at the highest dose, at which final body weight was 17% lower in the moxonidine-treated animals compared with controls. The moxonidine-treated (6 and 10 mg/kg) obese animals had significantly lower fasting plasma levels of insulin (17% and 19%, respectively) and free fatty acids (36% and 28%, respectively), whereas plasma glucose was not altered. During an oral glucose tolerance test, the glucose response (area under the curve) was 47% and 67% lower, respectively, in the two highest moxonidine-treated obese groups. Moreover, glucose transport activity in the isolated epitrochlearis muscle stimulated by a maximally effective insulin dose (13.3 nmol/L) was 39% and 70% greater in the 6 and 10 mg/kg moxonidine-treated groups, respectively (P<.05 for all effects). No significant alterations in muscle glucose transport were elicited by 2 mg/kg moxonidine. These findings indicate that in the severely insulin-resistant and dyslipidemic obese Zucker rat, chronic in vivo treatment with moxonidine can significantly improve, in a dose-dependent manner, whole-body glucose tolerance, possibly as a result of enhanced insulin-stimulated skeletal muscle glucose transport activity and reduced circulating free fatty acids. PMID- 9403584 TI - Dopamine D3 receptor in peripheral mononuclear cells of essential hypertensives. AB - Dopamine D3 receptor was studied in peripheral mononuclear cells of high-normal, stage 1, stage 2, and stage 3 essential hypertensives using a radioligand binding assay technique with [3H]-7-hydroxy-N,N-di-n-propyl-2-aminotetraline (7-OH-DPAT) as a radioligand. A group of de novo Parkinsonian patients was also examined as a reference group of impaired dopaminergic function. [3H]-7-OH-DPAT was bound specifically to human peripheral mononuclear cells in a manner consistent with the labeling of a dopamine D3 receptor. No changes in free dopamine, norepinephrine, epinephrine and aldosterone levels, renin activity, dissociation constant of [3H]-7-OH-DPAT binding, or the pharmacological profile of [3H]-7-OH DPAT binding were found between normotensive control subjects and essential hypertensives or Parkinsonians. The density of peripheral mononuclear cell [3H]-7 OH-DPAT binding sites increased in essential hypertensives parallel to blood pressure value augmentation. A higher density of [3H]-7-OH-DPAT binding sites was found in Parkinsonians. In these patients, the density of [3H]-7-OH-DPAT binding sites was similar to that observed in high-normal subjects and in stage 1 essential hypertensives. The increased density of peripheral mononuclear cell dopamine D3 receptor in hypertension as well as in Parkinson's disease may represent an upregulation mechanism consequent to impaired dopaminergic function. In view of the difficulty in identifying markers of peripheral dopamine function, analysis of dopamine D3 receptor in peripheral mononuclear cells may help evaluate whether the dopaminergic system is involved in hypertension. PMID- 9403585 TI - Patterns of neuronal activation during development of sodium sensitive hypertension in SHR. AB - The effects of regular (RNa) or high (HNa) sodium diet for 3, 7, and 14 days on Fra-like immunoreactivity (Fra-LI) in the brains of Wistar-Kyoto rats (WKY) and spontaneously hypertensive rats (SHR) were examined using an antibody that recognizes all known members of the Fos family (Fos, Fos-B, Fra-1, and Fra-2). Two weeks of HNa significantly exacerbated hypertension in SHR but had no effects in WKY. On RNa, compared with WKY, SHR showed higher Fra-LI in the median preoptic nucleus, supraoptic nucleus, both parts of the paraventricular nucleus, nucleus of the solitary tract, and central gray. Fra-LI in the subfornical organ did not differ between the two strains. On RNa, Fra-LI in the anterior hypothalamic area could be detected only in WKY. In osmoregulatory areas, HNa diet increased Fra-LI in both SHR and WKY to comparable extents, but in the median preoptic nucleus, Fra-LI was increased to a greater extent in SHR. HNa produced smaller increases in the subfornical organ of SHR compared with WKY. In both the parvocellular and magnocellular paraventricular nuclei, increases in Fra LI by HNa were more pronounced in SHR than in WKY. In the anterior hypothalamic area, Fra-LI could no longer be detected in WKY on HNa, whereas it appeared in SHR. HNa increased Fra-LI in the NTS and central gray to similar levels in WKY and SHR. These results indicate that WKY and SHR differ in the pattern of neuronal activation accompanying maturation on RNa. HNa activates neurons in a number of brain areas, and the pattern of these changes also differs between WKY and SHR. PMID- 9403586 TI - Role of endothelin in hypertension of experimental chronic renal failure. AB - Surgical ablation of renal mass leads to a reduction in kidney function and commonly to the development of hypertension and chronic renal failure (CRF) in rats. The objective of this study was to determine whether endothelin (ET)-1 is involved in the maintenance of the hypertension that accompanies loss of renal mass. First, we demonstrated the antihypertensive efficacy of PD 155080, a selective, orally active ET(A) receptor antagonist, in a group of rats made hypertensive by continuous intravenous infusion of ET-1 (2.5 pmol x kg(-1) x min[ 1]) for 7 days. ET-1 produced a sustained hypertension and PD 155080 (56.4 micromol/kg [25mg/kg] BID PO) normalized blood pressure (BP) during the 5 days of drug administration. In a second experiment, Sprague-Dawley rats underwent a 5/6 reduction in renal mass (RRM); 4 weeks later, PD 155080 administered for 7 days resulted in a sustained reduction in BP. Sham-operated rats also showed a slight hypotensive response to PD 155080 administration. Plasma urea nitrogen, plasma creatinine, urinary protein excretion, and creatinine clearance were not altered by PD 155080 administration in RRM or sham rats. In a third experiment, we investigated the contribution of the renin-angiotensin system to BP control in RRM rats given PD 155080. In these rats, PD 155080 reduced BP during 5 treatment days, and this antihypertensive effect was not altered by coadministration of the angiotensin-converting enzyme inhibitor enalapril in the drinking water (508 micromol/L [250 mg/L]). These results demonstrate that (1) ET-1 plays a role in established RRM hypertension through activation of the ET(A) receptor subtype, (2) lowering blood pressure with PD 155080 in RRM rats does not adversely affect renal function, and 3) the antihypertensive effect of ET(A) receptor antagonism is not opposed by the renin-angiotensin system. PMID- 9403587 TI - Effects of continuous infusion of endothelin-1 in pregnant sheep. AB - Plasma concentration of endothelin-1, a potent vasoconstrictor produced by the vascular endothelium, has been observed to be significantly increased in a number of pathophysiological states, including preeclampsia. In the present study we have evaluated the effects of elevated plasma endothelin-1 in pregnant sheep by continuous exogenous endothelin-1 administration. Nine pregnant ewes (110+/-5 days' gestation) were instrumented for measurements of maternal mean arterial pressure, renal blood flow, and uterine blood flow. After recovery, endothelin-1 was infused intravenously for 4 hours at a dose that was adjusted to raise mean arterial pressure by approximately 20 mm Hg by the end of the first hour (range 5 to 20 ng/kg per minute). Mean arterial pressure, renal blood flow, uterine blood flow, urinary protein excretion, hematocrit, and plasma endothelin-1 concentration were measured hourly, and renal and uterine vascular resistances were calculated. Endothelin-1 produced significant increases (% change from baseline at t=4 hours) in mean arterial pressure (45+/-8%), renal vascular resistance (353+/-66 %), and uterine vascular resistance (59+/-21%). Endothelin-1 also increased microvascular permeability both systemically and within the kidney, as suggested by marked increases in hematocrit (0.27+/-0.01 to 0.32+/ 0.01) and urinary protein concentration (0.95+/-0.1 to 7.9+/-3.2 mg/mL per mg creatinine). There was a highly significant correlation (P<.0001) between plasma endothelin-1 and mean arterial pressure, renal vascular resistance, uterine vascular resistance, hematocrit, and urinary protein content in all sheep studied. In addition, plasma endothelin-1 corresponded well with the time course of the changes in cardiovascular parameters and urinary protein excretion observed. These results provide evidence to suggest that elevation of circulating endothelin-1 in pregnant sheep can produce cardiovascular and hemodynamic changes that in many ways resemble the human disease preeclampsia. This supports the hypothesis that endothelial cell damage and/or dysfunction that is associated with increased production of endothelin-1 could directly contribute to the progression of preeclampsia. PMID- 9403588 TI - Endothelin converting enzyme-1 gene expression in the kidney of spontaneously hypertensive rats. AB - Abnormal renal handling of water and sodium is implicated in the pathogenesis of hypertension in spontaneously hypertensive rats (SHR). Alteration of renal endothelin-1 synthesis is also reported in SHR. Endothelin-1, a potent vasoconstrictor and regulator of sodium reabsorption in the nephron, has a pathophysiological potential in the development of hypertension. Because synthesis of bioactive endothelin-1 requires endothelin converting enzyme-1 (ECE 1), we investigated whether renal ECE-1 gene expression is altered in the kidney of SHR. Kidneys from both 4- and 12-week-old SHR and age-matched Wistar-Kyoto rats (WKY) were studied. ECE-1 mRNA in microdissected nephron segments was assessed by reverse transcription-competitive polymerase chain reaction, and ECE 1 protein level by Western blot. In 4-week-old SHR, ECE-1 mRNA was significantly increased in the proximal straight tubule, medullary thick ascending limb, cortical thick ascending limb, and inner medullary collecting duct. ECE-1 protein level was increased in both the outer and inner medulla. In 12-week-old SHR, ECE 1 gene expression was significantly increased in the proximal straight tubule, medullary thick ascending limb, and also in the glomeruli. Glomerular preproendothelin-1 mRNA expression was not different between the two strains at both 4 and 12 weeks. We conclude that high ECE-1 gene expression in the nephron, via increase of endothelin-1 synthesis, may promote sodium retention that contributes to the development and/or maintenance of hypertension in SHR. PMID- 9403589 TI - Endothelium-derived hyperpolarizing factor, but not nitric oxide, is reversibly inhibited by brefeldin A. AB - The subcellular localization of the enzymes synthesizing endothelium-derived vasodilator autacoids has been proposed to play a role in determining the ability of endothelial cells to enhance autacoid production in response to stimulation. We therefore investigated the effects of brefeldin A-induced disruption of the Golgi apparatus and Golgi-plasma membrane trafficking on the production of nitric oxide (NO), prostacyclin, and the endothelium-derived hyperpolarizing factor (EDHF) by native and cultured endothelial cells. In porcine coronary artery segments, brefeldin A (35 micromol/L, 90 minutes) did not affect relaxations to sodium nitroprusside or the K+ channel opener cromakalim but elicited a rightward shift in the concentration-response curve to bradykinin without altering the maximum vasodilator response (Rmax). Brefeldin A failed to attenuate the bradykinin-induced, NO-mediated relaxation under depolarizing conditions but inhibited the bradykinin response under conditions of combined cyclooxygenase/NO synthase blockade, suggesting that this agent selectively interferes with the production of EDHF. Indeed, incubation of porcine coronary arteries with brefeldin A, which did not affect the bradykinin-induced accumulation of either cyclic GMP or 6-keto-prostaglandin F1alpha, markedly and reversibly attenuated the EDHF-mediated hyperpolarization of detector smooth muscle cells in a patch clamp bioassay system. The microtubule destabilizer nocodazole also affected both the EC50 and Rmax to bradykinin in porcine coronary arteries. Since EDHF is thought to be a cytochrome P450-derived metabolite of arachidonic acid and both brefeldin A and nocodazole are known to interfere with the targeting of cytochrome P450 from the Golgi apparatus to the plasma membrane, it is conceivable that brefeldin A inhibits EDHF formation by preventing the targeting of the EDHF-synthesizing enzymes to the plasma membrane. PMID- 9403590 TI - Lacidipine restores endothelium-dependent vasodilation in essential hypertensive patients. AB - Essential hypertension is characterized by impaired endothelium-dependent vasodilation. The present study was designed to test whether antihypertensive treatment with the calcium antagonist lacidipine can improve endothelium dependent vasodilation in essential hypertensive patients. In 12 normotensive subjects (mean age, 47.8+/-8.6 years; blood pressure, 118.6+/-4.2/76.7+/-3.9 mm Hg) and 19 hypertensive patients (mean age, 49.4+/-10.2 years; blood pressure; 153.5+/-13.3/101.3+/-6.4 mm Hg), we studied forearm blood flow modifications (strain-gauge plethysmography) induced by intrabrachial infusion of acetylcholine (0.15, 0.45, 1.5, 4.5, and 15 microg/100 mL per minute) and bradykinin (5, 15, and 50 ng/100 mL per minute), two endothelium-dependent vasodilators that act through different receptors and signal transduction pathways, and sodium nitroprusside (1, 2, and 4 microg/100 mL per minute), an endothelium-independent vasodilator. In essential hypertensive patients, vascular reactivity was repeated during prolonged (8 weeks of oral treatment at 6 mg/d) lacidipine administration and 2 weeks after withdrawal of chronic (32-week) treatment. Hypertensive patients showed significantly (P<.01) blunted vasodilation in response to acetylcholine (vascular resistance, 31.5+/-4.9 to 7.6+/-2.4 SU) and bradykinin (vascular resistance, 32.3+/-5.8 to 8.5+/-3.0 SU) compared with control subjects (vascular resistance: acetylcholine, 24.3+/-3.9 to 3.7+/-1.2 SU; bradykinin, 24.7+/-0.4 to 4.1+/-1.3 SU), whereas the response to sodium nitroprusside was similar. After either 8 or 32 weeks of lacidipine treatment, the vasodilation in response to acetylcholine (30.6+/-7.7 to 5.7+/-1.5 and 34.3+/-6.6 to 5.9+/-1.9 SU, respectively) and bradykinin (31.3+/-7.2 to 6.4+/-1.6 and 33.7+/-5.4 to 6.1+/ 1.5 SU, respectively), but not to sodium nitroprusside, proved to be significantly (P<.05) increased compared with baseline. In essential hypertensive patients, oral treatment with lacidipine increased forearm vasodilation in response to acetylcholine and bradykinin, suggesting that this drug can improve endothelial function in patients with essential hypertension. PMID- 9403591 TI - Bosentan prevents preglomerular alterations during angiotensin II hypertension. AB - The present study was performed to characterize structurofunctional alterations of preglomerular vessels during chronic angiotensin II (Ang II)-induced hypertension (Ang II group: 400 ng x kg[-1] x min[-1], 10 days) and to assess the role of endothelin-1 in rats receiving Ang II and the mixed receptor antagonist bosentan (Ang II+B group: 30 mg x kg[-1] x d[-1], 10 days). Systolic blood pressure rose by 56+/-3 and 54+/-6 mm Hg in Ang II and Ang II+B rats, respectively. Albuminuria increased similarly in both Ang II-treated groups, reflecting glomerular barrier dysfunction. Preglomerular vessels were isolated after HCI maceration and comprised arcuate arteries and their branches, interlobular arteries (ILA), and afferent arterioles (AA). In the Ang II group, focal vascular lesions affected 36+/-6%, 20+/-5%, and 4+/-1% of arcuate arterial branches, ILA, and AA, respectively. They were characterized by 74% increased media thickness and accumulation of Sudan black-positive (SB+) lipid droplets, and media cell proliferation was documented through immunohistochemistry. The occurrence of SB+ lesions was strikingly reduced with bosentan. Autoregulatory responses (AR) were assessed along ILA and AA with the use of blood-perfused juxtamedullary nephron preparations. AR were elicited by raising blood perfusion pressure from 60 to 160 mm Hg and quantified through videomicroscopy as pressure induced constrictions. AR were inhibited in Ang II-treated rats along ILA and AA; Ang II-induced AR changes were prevented by bosentan. Maximal relaxation induced by Mn2+ revealed equal basal tone in Ang II-treated, Ang II+B-treated, and control vessels. Chronic Ang II-induced hypertension is therefore associated with the development of SB+ lesions and selective impairment of AR in juxtamedullary nephrons. Endothelin-1 likely mediates the structurofunctional alterations of preglomerular vasculature during Ang II hypertension. PMID- 9403592 TI - Chronic angiotensin-converting enzyme inhibition and angiotensin II type 1 receptor blockade: effects on cardiovascular remodeling in rats induced by the long-term blockade of nitric oxide synthesis. AB - We have shown previously that angiotensin-converting enzyme (ACE) inhibitors prevent coronary vascular remodeling (medial thickening and perivascular fibrosis) and myocardial remodeling (fibrosis and hypertrophy) in rats induced by long-term inhibition of nitric oxide (NO) synthesis with oral administration of N omega-nitro-L-arginine methyl ester (L-NAME). ACE inhibitors inhibit both the formation of angiotensin II and the catabolism of bradykinin. In this study, we aimed to determine the relative contribution of the latter two mechanisms to the beneficial effects of an ACE inhibitor on structural remodeling. First, we examined the effects of the ACE inhibitor temocapril and the angiotensin II AT1 subtype receptor antagonist CS-866 on the structural remodeling induced by administering L-NAME for 8 weeks. Temocapril and CS-866 were equally effective in preventing remodeling. Second, we examined whether the effect of temocapril on the remodeling induced by L-NAME was reduced by the bradykinin receptor antagonist HOE140. The latter drug did not alter the beneficial effect of temocapril on remodeling. In conclusion, although species differences must be considered to apply our conclusion to clinical conditions, the present results suggest that the inhibition of angiotensin II activity, mediated via the AT1 receptors, is responsible for the beneficial effects of an ACE inhibitor in our animal model of coronary vascular and myocardial remodeling induced by the long term inhibition of NO synthesis. PMID- 9403593 TI - Oxidative stress in the Dahl hypertensive rat. AB - Enhanced production of oxygen free radicals may play a role in hypertension by affecting vascular smooth muscle contraction, resistance to blood flow, and organ damage. The aim of this study was to determine whether oxygen free radicals are involved in the development of salt-induced hypertension. Dahl salt-sensitive (Dahl-S) and salt-resistant (Dahl-R) rats were fed either a high salt (6.0% NaCl) or low salt (0.3% NaCl) diet for 4 weeks. The high salt diet caused the development of severe hypertension in Dahl-S animals and had no effect on blood pressure in Dahl-R animals. A tetranitroblue tetrazolium dye was used to detect superoxide radicals in microvessels of the mesentery. Light absorption measurements revealed enhanced staining along the endothelium of arterioles and venules in hypertensive Dahl-S animals, with significantly lower values in normotensive animals. In addition, a Clark electrochemical electrode was used to measure hydrogen peroxide levels in fresh plasma. Hypertensive Dahl-S animals had a higher plasma hydrogen peroxide concentration compared with their normotensive counterparts (2.81+/-0.43 versus 2.10+/-0.41 micromol/L), while no difference was detected between high- and low salt-treated Dahl-R animals (1.70+/-0.35 versus 1.56+/-0.51 micromol/L). The plasma hydrogen peroxide levels of all groups correlated with mean arterial pressure (r=.77). These findings demonstrate an enhanced production of oxygen free radicals in the microvasculature of hypertensive Dahl-S rats. PMID- 9403594 TI - Fatty acids augment endothelium-dependent dilation in hand veins by a cyclooxygenase-dependent mechanism. AB - Evidence supports the hypothesis that elevated nonesterified fatty acids (NEFAs) in patients with insulin resistance, eg, obese hypertensive subjects, contribute to increased vascular alpha-adrenergic reactivity and tone by impairing endothelium-dependent vasodilation. To generate further support for this notion, we studied responses to endothelium-dependent and independent dilators under control (0.9% NaCl/heparin) conditions in one hand and with elevated NEFAs in the contralateral hand (10% intralipid/heparin). To observe venodilator responses, the dorsal hand vein diameter was first reduced by approximately 60% with phenylephrine. Studies were repeated with indomethacin to block the generation of cyclooxygenase products. In contrast to previous in vitro data, elevating NEFAs locally in vivo augmented rather than suppressed venodilator responses to the two endothelium-dependent dilators acetylcholine and methacholine (P<.05). Responses to the endothelium-independent dilator nitroglycerin were unaffected. Indomethacin attenuated the capacity of intralipid/heparin to enhance endothelium dependent dilator responses to acetylcholine and methacholine. Indomethacin did not affect venodilator responses to nitroglycerin. The effect of intralipid/heparin to significantly reduce the phenylephrine infusion rate required to reduce hand vein diameter by approximately 60% was reversed by indomethacin. These data indicate that raising fatty acids locally augments endothelium-dependent dilation by a cyclooxygenase-dependent mechanism. The findings also suggest that NEFAs augment alpha-adrenoceptor-mediated constriction in hand veins by a cyclooxygenase-dependent mechanism. These hand vein studies do not support the notion that the elevated NEFAs in obese hypertensive patients augment alpha1-adrenoceptor-mediated reactivity by reducing nitric oxide synthesis. PMID- 9403595 TI - Influence of family history on frequency of glucagon receptor Gly40Ser mutation in hypertensive subjects. PMID- 9403596 TI - Essential hypertension and insulin clearance and resistance. PMID- 9403597 TI - Cancer risk in users of calcium channel blockers. PMID- 9403598 TI - Cancer risk in users of calcium channel blockers. PMID- 9403599 TI - Trends in cardiovascular mortality in Europe. PMID- 9403600 TI - Heart research: celebration and renewal. PMID- 9403601 TI - Transition from hypertrophy to failure. PMID- 9403602 TI - Monitoring platelet GP IIb/IIIa [corrected] antagonist therapy. PMID- 9403603 TI - GATA4: a novel transcriptional regulator of cardiac hypertrophy? PMID- 9403604 TI - Population versus clinical views in coronary disease: can epidemiological data be useful to clinicians? PMID- 9403605 TI - Aortic plaque morphology and vascular events: a follow-up study in patients with ischemic stroke. FAPS Investigators. French Study of Aortic Plaques in Stroke. AB - BACKGROUND: Atherosclerotic disease of the aortic arch has been found to be associated with the risk of ischemic stroke. We have shown that atherosclerotic plaques > or = 4 mm in thickness in the ascending aorta and proximal arch detected by transesophageal echocardiography are a risk factor for ischemic stroke. The purpose of this study was to evaluate the impact, if any, of plaque morphology (ulceration, hypoechoic plaques or calcification) on the risk of subsequent vascular events. METHODS AND RESULTS: We followed for a period of 2 to 4 years, a cohort of 334 patients 60 years or older who were consecutively admitted with brain infarction and who had transesophageal echocardiography. The risk of vascular events in patients with plaques in the aortic arch according to the presence of surface ulceration, calcifications, and sessile or mobile thrombus was estimated during a total of 788 person-years of follow-up. Hypoechoic plaques, calcifications, and ulceration were more frequently found in patients with plaques > or = 4 mm as compared with those with plaques < 4 mm. The presence of ulceration did not increase the relative risk of vascular events in patients with plaque > or = 4 mm (the relative risk was 4.3 [P<.001] in those with ulceration and 5.7 [P<.001]) in those without ulceration. The lack of calcification did increase the risk of vascular events in patients with plaque > or = 4 mm. The highest relative risk of events was found among the patients with noncalcified plaques (relative risk, 10.3; 95% confidence interval, 4.2 to 25.2; P<.001). The risk of events was systematically higher in patients without calcifications than in patients with calcifications regardless of what other morphological features were considered. CONCLUSIONS: In patients with brain infarction, the risk associated with aortic plaque thickness (> or = 4 mm) is markedly increased by the absence of plaque calcifications. These findings are important for the design of therapeutic trials in such patients. PMID- 9403606 TI - Exaggerated blood pressure responses during mental stress are associated with enhanced carotid atherosclerosis in middle-aged Finnish men: findings from the Kuopio Ischemic Heart Disease Study. AB - BACKGROUND: Exaggerated cardiovascular reactivity to mental stress is hypothesized to increase atherosclerotic risk. We examined this hypothesis using cross-sectional data from the Kuopio Ischemic Heart Disease study, a population based epidemiological sample. METHODS AND RESULTS: 901 Eastern Finnish men from four age cohorts (age, 42 to 60 years) were administered a standardized testing battery to assess cardiovascular reactivity to mental stress. Ultrasound measures of intima-medial thickness (IMT) and plaque height from the common carotid arteries were used as noninvasive markers of atherosclerosis. Diastolic blood pressure (DBP) responses to mental stress were significantly associated with mean IMT (b=.021, P=.006), maximum IMT (b=.026, P=.013), and mean plaque height (b=.017, P=.041). Significant associations were also shown between stress-related systolic blood pressure (SBP) reactivity and mean IMT (b=.0151, P=.042). When examined separately by age, associations with IMT were significant only in the youngest half of the sample (age, 46 and 52 years, n=433; for mean IMT, DBP b=.033, P=.0002, SBP b=.0266, P=.003; for maximum IMT, DBP b=.039, P=.002, SBP b=.032, P=.011). Results remained significant in the younger subjects after adjustment for smoking, lipid profiles, fasting glucose, and resting blood pressure (b=.024, P=.011); results also remained significant in a subgroup of unmedicated younger subjects without symptomatic cardiovascular disease (n=135; for SBP reactivity, b=.031, P=.036; for DBP, b=.037, P=.007). CONCLUSIONS: The tendency to show exaggerated pressor responses to mental stress is a significant independent correlate of atherosclerosis in this population sample of Finnish men. The effect does not appear to be accounted for by the confounding influence of other risk factors or preexisting clinical disease. PMID- 9403607 TI - Population versus clinical view of case fatality from acute coronary heart disease: results from the WHO MONICA Project 1985-1990. Multinational MONItoring of Trends and Determinants in CArdiovascular Disease. AB - BACKGROUND: The clinical view of case fatality (CF) from acute myocardial infarction (AMI) in those reaching the hospital alive is different from the population view. Registration of both hospitalized AMI cases and out-of-hospital coronary heart disease (CHD) deaths in the WHO MONICA Project allows both views to be reconciled. The WHO MONICA Project provides the largest data set worldwide to explore the relationship between CHD CF and age, sex, coronary event rate, and first versus recurrent event. METHODS AND RESULTS: All 79,669 events of definite AMI or possible coronary death, occurring from 1985 to 90 among 5,725,762 people, 35 to 64 years of age, in 29 MONICA populations are the basis for CF calculations. Age-adjusted CF (percentage of CHD events that were fatal) was calculated across populations, stratified for different time periods, and related to age, sex, and CHD event rate. Median 28-day population CF was 49% (range, 35% to 60%) in men and 51% (range, 34% to 70%) in women and was particularly higher in women than men in populations in which CHD event rates were low. Median 28-day CF for hospitalized events was much lower: in men 22% (range, 15% to 36%) and in women 27% (range, 19% to 46%). Among hospitalized events CF was twice as high for recurrent as for first events. CONCLUSIONS: Overall 28-day CF is halved for hospitalized events compared with all events and again nearly halved for hospitalized 24-hour survivors. Because approximately two thirds of 28-day CHD deaths in men and women occurred before reaching the hospital, opportunities for reducing CF through improved care in the acute event are limited. Major emphasis should be on primary and secondary prevention. PMID- 9403608 TI - Rapid assessment of platelet function with a modified whole-blood aggregometer in percutaneous transluminal coronary angioplasty patients receiving anti-GP IIb/IIIa therapy. AB - BACKGROUND: The glycoprotein (GP) IIb/IIIa receptor antagonist abciximab (c7E3 Fab, ReoPro) is approved for use in high-risk percutaneous transluminal coronary angioplasty (PTCA). At present, no "point of care" exists for measuring pharmacological GP IIb/IIIa blockade. To address this need, the Chrono-log Whole Blood Aggregometer, which measures platelet aggregation by electrical impedance, was adapted to test platelet function at the bedside. METHODS AND RESULTS: GP IIb/IIIa receptor blockade, impedance (5 microg/mL collagen), and turbidimetric aggregation (5 and 20 micromol/L ADP) measurements were obtained on 14 PTCA patients who received the standard bolus plus a 12-hour infusion of abciximab. During abciximab administration, mean GP IIb/IIIa receptor blockade was > 91%, and both impedance and turbidimetric aggregation were inhibited by > or = 90%. At 12 hours after abciximab treatment, the mean inhibition of turbidimetric platelet aggregation to 5 and 20 micromol/L ADP was 65+/-20% and 49+/-14%, respectively, and inhibition of impedance aggregation was 69+/-12%. GP IIb/IIIa receptor blockade was 67+/-8%. At 36 hours after abciximab treatment (n=8), the mean inhibition of turbidimetric platelet aggregation to 5 and 20 micromol/L ADP was 44+/-21% and 30+/-14%, respectively, whereas impedance aggregation was inhibited by 60+/-14%. GP IIb/IIIa receptor blockade was 57+/-7%. CONCLUSIONS: During and at 12 hours after abciximab therapy, impedance and turbidimetric platelet aggregation to 5 micromol/L ADP were comparable and closely correlated with GP IIb/IIIa receptor blockade. However, at 36 hours after abciximab treatment, impedance platelet aggregation more closely paralleled GP IIb/IIIa receptor blockade and indicated a slower recovery of platelet function than turbidimetric aggregometry. PMID- 9403609 TI - Contemporary percutaneous treatment of unprotected left main coronary stenoses: initial results from a multicenter registry analysis 1994-1996. AB - BACKGROUND: Coronary artery bypass surgery (CABG) has been considered the therapy of choice for patients with unprotected left main (ULMT) coronary stenoses. Selected single-center reports suggest that the results of percutaneous intervention may now approach those of CABG. METHODS AND RESULTS: To assess the results of percutaneous ULMT treatment from a wide variety of experienced interventional centers, we requested data on consecutive patients treated after January 1, 1994, from 25 centers. One hundred seven patients were identified who were treated either electively (n=91) or for acute myocardial infarction (n=16). Of patients treated electively, 25% were considered inoperable, and 27% were considered high risk for bypass surgery. Primary treatment included stents (50%), directional atherectomy (24%), and balloon angioplasty (20%). Follow-up was 98.8% complete at 15+/-8 months. Results varied considerably, depending on presentation and treatment. For patients with acute myocardial infarction, technical success was achieved in 75%, and survival to hospital discharge was 31%. For elective patients, technical success was achieved in 98.9%, and in-hospital survival was strongly correlated with left ventricular ejection fraction (P=.003). Longer-term event (death, infarction, or bypass surgery) -free survival was correlated with ejection fraction (P<.001) and was inversely related to presentation with progressive or rest angina (P<.001). Surgical candidates with ejection fractions > or = 40% had an in-hospital survival of 98% and a 9-month event-free survival of 86+/-5%, whereas patients with ejection fractions < 40% had 67% and 22+/-12% in-hospital and 9-month event-free survivals, respectively. Nine hospital survivors (10.6%) experienced cardiac death within 6 months of hospital discharge. CONCLUSIONS: While results for selected patients appear promising, until early post-hospital discharge cardiac death can be better understood and minimized, percutaneous revascularization of ULMT stenosis should not be considered an alternative to bypass surgery for most patients. When percutaneous revascularization of ULMT is required, directional atherectomy and stenting appear to be the preferred techniques, and follow-up angiography 6 to 8 weeks after treatment is probably advisable. PMID- 9403610 TI - Long-term angiographic and clinical outcome of patients undergoing multivessel coronary stenting. AB - BACKGROUND: Randomized clinical trials have shown that multivessel coronary angioplasty is feasible and provides similar long-term survival as bypass surgery in selected patients. However, the higher need for repeat intervention, in particular, coronary artery bypass graft surgery, remains a problem. The objective of this study was to test the hypothesis that multivessel stenting is safe and effective in reducing the need for repeat interventions, in particular, the need for bypass surgery. METHODS AND RESULTS: Between March 1993 and June 1995, 100 consecutive patients (243 lesions) had multivessel coronary stenting. High-pressure stent optimization was used in all patients. Procedural success was achieved in 97% of lesions; 2 patients (2%) required emergency bypass surgery. Angiographic follow-up was obtained in 89% of patients at 5.2+/-2.5 months. Angiographic restenosis occurred in 22% of the lesions, but 37% of patients had > or = 1 lesion with restenosis. Clinical follow-up was obtained in all patients at 21 +/- 10 months: target lesion revascularization was needed in 30 patients (30%), repeat angioplasty in 28 patients (28%) and coronary bypass surgery in 2 patients (2%); the overall survival rate was 96% (2% noncardiac death). CONCLUSIONS: Multivessel coronary stenting can be performed with high success rate and low need for emergency bypass surgery. Compared with historical results with multivessel percutaneous transluminal coronary angioplasty, patients who undergo multivessel stenting need less repeat interventions, in particular, less coronary bypass surgery and have similar long-term survival. PMID- 9403611 TI - Bimodal distribution of angiographic measures of restenosis six months after coronary stent placement. AB - BACKGROUND: Restenosis has been perceived as the tail end of a normal distribution of the response of the vessel to the intervention. However, recent studies have described a bimodal distribution for de novo lesions after percutaneous transluminal coronary angioplasty. This finding suggests that some lesions may be more susceptible for restenosis. Whether this holds true for a wider spectrum of lesions undergoing stent placement is not yet known. The present study analyzes the frequency distribution of angiographic indexes of restenosis 6 months after coronary stent implantation. METHODS AND RESULTS: Quantitative angiographic evaluation was performed in 1084 lesions of 1084 patients before, immediately after, and 6 months after successful Palmaz-Schatz stent placement; this represented 80.4% of patients eligible for follow-up angiography. Principal end points of the analysis were angiographic indexes of restenosis at 6 months. Twenty-two lesions that became totally occluded at follow up were excluded from most parts of the analysis. Diameter stenosis, minimal luminal diameter (MLD), and lumen loss at 6 months did not follow a normal pattern; the bimodal pattern was demonstrated through deconvolution that yielded two separate normal components delineating two lesion populations, which developed distinctively different degrees of lumen renarrowing. The first and larger subgroup of lesions, which was less prone to restenosis, was centered around a mean value of 27% for diameter stenosis and 2.19 mm for MLD, whereas the second subgroup, with a greater tendency for restenosis, was situated around a mean value of 68% for diameter stenosis and 0.76 mm for MLD. The intersection point between the two theoretical normal distribution components was 53.5% for diameter stenosis and 1.09 mm for MLD at follow-up. CONCLUSIONS: Frequency distribution curves of angiographic indexes of restenosis after coronary stent placement have a bimodal pattern, suggesting the existence of two distinct populations with different propensity to restenosis. These findings may encourage future efforts for the timely identification of the subset with a higher risk as the target of specific antirestenotic strategies. PMID- 9403612 TI - Aggregating human platelets stimulate the expression of thrombin receptors in cultured vascular smooth muscle cells via the release of transforming growth factor-beta1 and platelet-derived growth factorAB. AB - BACKGROUND: Thrombin and the thrombin receptor have been implicated in the proliferation of vascular smooth muscle cells (VSMCs) observed after angioplasty and in atherosclerosis. Because thrombin receptor activation is an irreversible proteolytic event, the marked upregulation of the smooth muscle cell thrombin receptor after vascular injury may account for the maintained mitogenic activity of thrombin. The present study was designed to determine whether aggregating platelets stimulate thrombin receptor expression in cultured VSMCs and, if so, to identify the mediators. METHODS AND RESULTS: Thrombin receptor expression was assessed by Northern and Western blot analyses and functionally by measuring the release of 6-keto prostaglandin F1alpha. Platelet-derived products (PDPs) released by aggregating human platelets enhanced thrombin receptor mRNA levels in a time- and concentration-dependent manner, an effect that was potentiated by transient acidification of PDPs, which release bioactive transforming growth factor (TGF)-beta1, and that was slightly inhibited by ketanserin. Among several factors known to be released by aggregating platelets, only TGF-beta1, platelet derived growth factorAB (PDGF(AB)), and serotonin mimicked the PDP effect. The level of membrane thrombin receptor protein was increased in TGF-beta1-treated VSMCs. Pretreatment of VSMCs with either acidified PDP, or TGF-beta1 increased the alpha-thrombin-stimulated release of 6-keto prostaglandin F1alpha. This effect was blunted by incubating acidified PDP with either a TGF-beta- or a PDGF neutralizing antibody. CONCLUSIONS: Aggregating human platelets stimulate the expression of thrombin receptors in VSMCs through the release of TGF-beta1, PDGF(AB), and, to a lesser extent, serotonin. The upregulation of the thrombin receptor by products released by aggregating platelets may sustain the mitogenic activity of thrombin in the vascular wall at sites of injury. PMID- 9403613 TI - Cardiovascular and sympathetic effects of nitric oxide inhibition at rest and during static exercise in humans. AB - BACKGROUND: Nitric oxide (NO) regulates vascular tone and blood pressure, and studies in animals suggest that it does so, at least in part, by modulating sympathetic neural outflow. Loss of NO-induced vasodilator tone and restraint on sympathetic vasoconstrictor outflow could lead to exaggerated vasoconstrictor and pressor responses to physical stress in humans. METHODS AND RESULTS: To determine the role of NO in the modulation of central sympathetic outflow and vascular tone at rest and during a physical stress, we tested effects of systemic inhibition of NO synthase by N(G)-monomethyl-L-arginine (L-NMMA) infusion (a stereospecific inhibitor of NO synthase) on sympathetic nerve activity (microneurography), regional vascular resistance, and blood pressure at rest and during static handgrip. The major new findings are that (1) under resting conditions, L-NMMA infusion, which increased mean arterial pressure by approximately 10%, did not have any detectable effect on muscle sympathetic nerve activity, whereas a similar increase in arterial pressure evoked by phenylephrine infusion (an NO independent vasoconstrictor) decreased the rate of sympathetic nerve firing by approximately 50%; (2) during static handgrip, the exercise-induced sympathetic nerve responses were preserved during L-NMMA infusion but markedly attenuated during phenylephrine infusion; and (3) the L-NMMA-induced loss of vasodilator tone did not result in exaggerated exercise-induced pressor and calf vasoconstrictor responses. CONCLUSIONS: These findings indicate that NO is involved in the central regulation of sympathetic outflow in humans and suggest that both neuronal and endothelial NO synthesis may contribute to the regulation of vasomotor tone. PMID- 9403614 TI - Three-dimensional mapping of the common atrial flutter circuit in the right atrium. AB - BACKGROUND: The full circuit of common atrial flutter using conventional methods of sequential or multielectrode activation mapping is not completely understood. METHODS AND RESULTS: We performed three-dimensional right atrial endocardial activation mapping during common counterclockwise atrial flutter in 17 patients (16 men, 1 woman; mean age, 53+/-11 years) by using the Cordis-Biosense EP Navigation system and assessed the distribution of estimated conduction velocities and double and fractionated potentials. ECG flutter wave morphologies were compared with activation patterns. Points (91+/-29) were sequentially acquired covering 88+/-11% of the flutter cycle length of 239+/-22 ms. A wide and variable posterior zone of double and fractionated potentials coincided with blocking and colliding wave fronts and formed the posterior limit of the circuit. A progressively widening septal (sep) wave front ascending from just beyond the coronary sinus ostium, passed cranially as a broad front anterior to the superior vena cava (SVC) in 14 patients, whereas fusion around the SVC formed the superior (sup) limb of the circuit in 3. Bounded anteriorly by the tricuspid valve, the wave front descended down the lateral (lat) aspect of the right atrium before completing the circuit in all cases through the inferior vena cava-tricuspid annulus isthmus. The estimated conduction velocity in the medial isthmus (0.6+/ 0.3 m/s) was lower than in the other limbs of the circuit (sup=1+/-0.5 m/s, lat=1+/-0.5 m/s, sep=0.9+/-0.4 m/s, P=.05). Double and fractionated potentials were constant and more prevalent in the posterior right atrium. ECG flutter wave morphology did not correlate with three-dimensional activation maps. CONCLUSIONS: Interindividual variations occur in the right atrial circuit of common atrial flutter, with constant activation through the cavotricuspid isthmus. A variable zone of block forms the posterior limit. Fusion around the SVC can occur, and ascending medial septal activation does not follow a consistent pattern. PMID- 9403615 TI - QT dispersion is determined by the extent of viable myocardium in patients with chronic Q-wave myocardial infarction. AB - BACKGROUND: QT dispersion is lower in patients with successful thrombolysis after acute myocardial infarction, suggesting that QT dispersion may be determined by the extent of viable and scarred myocardium. METHODS AND RESULTS: To test this hypothesis, QT dispersion was measured in a 12-lead resting ECG in 44 patients with chronic Q-wave myocardial infarction. To assess the extent of viable and scarred myocardium, all patients underwent F-18 fluorodeoxyglucose (FDG) positron emission tomography (PET). In addition, all patients had revascularization of the infarct-related artery and repeated angiography 4 months later. QT dispersion was lower (53+/-20 versus 94+/-24 ms, P<.0001) in patients with evidence of a substantial amount of viable myocardium in the infarct region as demonstrated by PET (average FDG uptake > or = 50% of normalized, maximum FDG uptake) than in patients with only minimal residual viability. Average FDG uptake of the infarct region and FDG defect size were significantly related to QT dispersion (r=.64, P<.0001; r=.67, P<.0001), whereas ejection fraction was not (r<.1, P=NS). QT dispersion of < or = 70 ms had a sensitivity of 85% and a specificity of 82% to predict viable myocardium in the infarct region. QT dispersion was also lower in patients with improvement of left ventricular function 4 months after revascularization (54+/-21 versus 88+/-30 ms, P=.0003). QT dispersion of < or = 70 ms had a sensitivity of 83% and a specificity of 71% to predict improvement of left ventricular function. CONCLUSIONS: QT dispersion is determined by the amount of viable myocardium in the infarct region and may serve as a novel, rapidly available marker of substantial viability in the infarct region of patients with chronic Q-wave myocardial infarction. PMID- 9403616 TI - Adenosine-induced atrioventricular block in patients with unexplained syncope: the diagnostic value of ATP testing. AB - BACKGROUND: ATP and its related nucleoside, adenosine, are ubiquitous biological compounds with potent depressant activity on the atrioventricular node. We hypothesized that an increased susceptibility of the atrioventricular node to adenosine may, in some cases, play a role in the genesis of syncope. METHODS AND RESULTS: The study was performed in two parts. In part 1, we evaluated the effects of a bolus injection of 20 mg ATP in a group of 60 patients (57+/-19 years, 31 men) with syncope of unexplained origin and in 90 control subjects without syncope (55+/-17 years, 46 men). In control subjects, the upper 95th percentile of the maximum RR interval distribution, during ATP-induced atrioventricular block (AVB), was 6000 ms. In the syncope group, 28% of patients had a maximum RR interval above this limit (P=.000). The distribution of the maximum RR interval below the 95th percentile was similar in the two groups. In part 2, we validated the ATP test in 24 patients who had the fortuitous ECG recording of a spontaneous syncope caused by a transient asystolic pause (AVB in 15 and sinus arrest in 9). The ATP test caused AVB with an asystolic pause of > or = 6000 ms in 53% of the patients with documented AVB but in none (0%) of the patients with documented sinus arrest (P=.01). Among the patients with spontaneous AVB, the ATP test was abnormal in 6 of the 7 patients (86%) in whom all conventional investigations for syncope had been negative and in 2 of the 8 patients (25%) who had shown positivity (P=.03). CONCLUSIONS: An increased susceptibility to ATP testing is present in patients with SUO and patients with syncope due to paroxysmal AVB. Thus, a logical inference is that ATP testing can be used to identify patients with syncope due to paroxysmal AVB. The results of this study form the necessary background for future prospective studies with an aim to validate this assumption. PMID- 9403617 TI - Interventional treatment for children with severe coronary artery stenosis with calcification after long-term Kawasaki disease. AB - BACKGROUND: About 4% of children with Kawasaki disease (KD) eventually develop ischemic heart disease, which is often associated with calcified stenosis. We assessed the utility of the percutaneous transluminal coronary rotational ablation (PTCRA) in children with coronary artery stenosis after KD. METHODS AND RESULTS: Four children (three boys and one girl; age, 12 to 13 years) with coronary artery stenosis underwent percutaneous transluminal coronary angioplasty (PTCA) and PTCRA 11.8+/-0.9 years after the onset of KD. Morphology of the coronary artery wall was evaluated by intravascular ultrasound imaging. In one patient, the targeted lesion for intervention was in the left anterior descending artery (90% stenosis); in the other three patients, it was in the middle of the right coronary artery (75% to 90% stenosis). PTCA failed in three patients because of severe stenosis with calcification. However, PTCRA proved effective, with stenosis rates reduced from 90% to 25%. Follow-up coronary angiography performed 4 months after the procedure demonstrated no restenosis, but mild aneurysms occurred in two patients. CONCLUSIONS: This study suggests that PTCRA is useful for revascularizing coronary arteries with severe stenosis and calcification as long-term sequelae of KD. Intravascular ultrasound imaging is useful in assessing the coronary artery wall pathology and in selecting the best treatment intervention. PMID- 9403618 TI - Cardiopulmonary interactions after Fontan operations: augmentation of cardiac output using negative pressure ventilation. AB - BACKGROUND: The low-output state is the chief cause of morbidity and mortality after Fontan operations. An alternative hemodynamic tool would be a welcome addition for these patients, who are typically resistant to conventional therapeutic measures. METHODS AND RESULTS: The hemodynamic effects of conversion from conventional intermittent positive pressure ventilation (IPPV) to cuirass negative pressure ventilation (NPV) was investigated in nine acute postoperative Fontan patients on the pediatric intensive care unit and nine anesthetized patients undergoing cardiac catheterization in the convalescent phase after Fontan operations. Pulmonary blood flow was measured using the direct Fick method during IPPV and after a brief period of NPV. In one subgroup of patients, pulmonary blood flow was measured again after reinstitution of IPPV, and in a second subgroup, pulmonary blood flow was measured after an extended period of NPV. A brief period of NPV increased pulmonary blood flow from 2.4 to 3.5 L x min(-1) x /m(-2), with a mean increase of 42%. Pulmonary blood flow continued to improve, with a total increase of 54% after an extended period of NPV. Values fell toward baseline after reinstitution of IPPV. Heart rate was unchanged during NPV, and the improvement in pulmonary blood flow was achieved by an increase in stroke volume from 25 mL/m2 to 37 mL/m2. CONCLUSIONS: Through improvement of the stroke volume alone, NPV brought about a marked increase in the pulmonary blood flow and, hence, cardiac output of Fontan patients. An improvement in cardiac output of this order, and by this mechanism, is currently unmatched by any therapeutic alternatives. PMID- 9403619 TI - cis-Acting sequences that mediate induction of beta-myosin heavy chain gene expression during left ventricular hypertrophy due to aortic constriction. AB - BACKGROUND: Marked alterations in the expression of specific genes occur during the development of cardiac hypertrophy in vivo. Little is known, however, about the cis-acting elements that mediate these changes in response to clinically relevant hypertrophic stimuli, such as hemodynamic overload, in intact adult animals. METHODS AND RESULTS: The left ventricular expression of a directly injected reporter gene driven by 3542 bp of rat beta-myosin heavy chain (beta MHC) promoter was increased 3.0-fold by aortic constriction (P<.005), an increment similar to the 3.2-fold increase in the level of the endogenous beta MHC mRNA in the same left ventricles. Subsequent analysis identified a 107-bp beta-MHC promoter sequence (-303/-197) sufficient to convert a heterologous neutral promoter to one that is activated by aortic constriction. These sequences contain two M-CAT elements, which have previously been demonstrated to mediate inducible expression during alpha1-adrenergic-stimulated hypertrophy in cultured neonatal cardiac myocytes, and a GATA element. Although simultaneous mutation of both M-CAT elements markedly decreased the basal transcriptional activity of an injected 333-bp beta-MHC promoter, it had no effect on aortic constriction stimulated transcription (3.5-fold increase, P<.005 for both wild type and mutant). In contrast, mutation of the GATA motif markedly attenuated aortic constriction-stimulated transcription (1.6-fold, P=NS) without affecting the basal transcriptional activity. This GATA site can interact with in vitro translated GATA-4 and compete with an established GATA site for GATA-4 binding activity in nuclear extracts from aortic constricted hearts. CONCLUSIONS: Basal and aortic constriction-stimulated transcription of the beta-MHC gene is mediated, at least in part, through different mechanisms. A GATA element within beta-MHC sequences -303/-197 plays a role in the transcriptional activation of this gene by aortic constriction. PMID- 9403620 TI - Angiotensin type 2 receptors are reexpressed by cardiac fibroblasts from failing myopathic hamster hearts and inhibit cell growth and fibrillar collagen metabolism. AB - BACKGROUND: Angiotensin (Ang) II type 1 receptor (AT1-R) induces cardiomyocyte hypertrophy and fibroblast proliferation, whereas the physiological role of AT2-R in cardiac remodeling remains poorly defined. METHODS AND RESULTS: Using Bio14.6 cardiomyopathic (CM) hamsters, we found that AT2-R sites were increased by 153% during heart failure compared with F1B controls. AT1-R numbers were increased by 72% in the hypertrophy stage and then decreased to the control level during heart failure. Such differential regulation of AT2-R and AT1-R during heart failure was consistent with changes in the respective mRNA levels. Autoradiography and immunocytochemistry revealed that both AT2-R and AT1-R are localized at higher densities in fibroblasts present in fibrous regions. Surrounding myocardium predominantly expressed AT1-R, but the level of expression was less than that in fibrous regions. Cardiac fibroblasts isolated from CM hearts during heart failure but not from control hamsters expressed AT2-R (30 fmol/mg protein). Using the cardiac fibroblasts expressing AT2-R, we found that Ang II stimulated net collagenous protein production by 48% and pretreatment with an AT2-R antagonist, PD123319, evoked a further elevation (83%). Ang II-induced synthesis of fibronectin and collagen type I were enhanced by 40% and 53%, respectively, by pretreatment with PD123319. Ang II-induced DNA synthesis (assessed by [3H]thymidine uptake) was significantly increased by PD123319, and the AT2-R agonist CGP42112A reduced the serum-stimulated increase in cell numbers by 23%. Treatment with an AT1-R antagonist, TCV116, for 20 weeks inhibited progression of interstitial fibrosis by 28%, whereas with 44-week PD123319 treatment but not 20 week treatment, the extent of the fibrous region was increased significantly, by 29%. CONCLUSIONS: These findings demonstrate that AT2-R is re-expressed by cardiac fibroblasts present in fibrous regions in failing CM hearts and that the increased AT2-R exerts an anti-AT1-R action on the progression of interstitial fibrosis during cardiac remodeling by inhibiting both fibrillar collagen metabolism and growth of cardiac fibroblasts. PMID- 9403621 TI - Chronic endothelin receptor blockade attenuates progressive ventricular dilation and improves cardiac function in rats with myocardial infarction: possible involvement of myocardial endothelin system in ventricular remodeling. AB - BACKGROUND: Left ventricular dilatation after myocardial infarction is associated with impaired ventricular function and heart failure and has important implications for survival. The purpose of this study was to investigate the role of endothelin-1 (ET-1) in ventricular dilatation and the effects of chronic endothelin receptor blockade by a mixed ET(A) and ET(B) receptor blocker (bosentan) on the circulating and cardiac endothelin systems. METHODS AND RESULTS: Three hours after coronary ligation or sham operation, bosentan (100 mg x kg body wt(-1) x d(-1)) or placebo was given by gavage. Seven days and 8 weeks after surgery, hemodynamic and left ventricular volume studies were performed. Acute bosentan treatment (7 days) had no effects on hemodynamic parameters and early left ventricular dilatation. In the rats with large infarcts, chronic bosentan treatment (8 weeks) versus placebo reduced left ventricular systolic pressure (116+/-2 versus 125+/-3 mm Hg, P<.05) and arterial pressure (93+/-2 versus 103+/-3 mm Hg, P<.05), improved stroke volume index (0.69+/-0.06 versus 0.52+/-0.04 mL/kg, P<.05), and prevented in part the rightward shift of the pressure-volume curve. Chronic bosentan treatment also decreased ET-1 levels (390+/-33 versus 475+/-22 pg/g tissue, P<.05) and density of ET-1 receptors (262+/-24 versus 346+/-31 fmol/mg protein, P<.05) in left ventricular myocardium. CONCLUSIONS: In the present study, a mixed ET(A) and ET(B) receptor antagonist (bosentan) partially prevented left ventricular dilatation and improved hemodynamics, suggesting that endothelin plays a role in left ventricular remodeling after myocardial infarction. Supporting this hypothesis, we show inhibitory effects of bosentan on the peripheral and myocardial endothelin system. PMID- 9403622 TI - Isolated myocyte contractile function is normal in postinfarct remodeled rat heart with systolic dysfunction. AB - BACKGROUND: Postinfarction ventricular remodeling is associated with lengthening and contractile dysfunction of the remote noninfarcted myocardium. Mechanisms underlying this phenomenon remain unclear. METHODS AND RESULTS: We studied serial changes in global left ventricular (LV) structure and function in infarcted (1, 2, 4, and 6 weeks after myocardial infarction) and sham-operated rat hearts and correlated them with structural and functional changes in myocytes isolated from the remote LV myocardium in the same hearts. Rats with myocardial infarction developed significant remodeling. The heart weight-to-body weight ratios were increased. LV volumes at filling pressure of 10 mm Hg were higher (305+/-28 versus 215+/-12 microL, P<.01). This was accompanied by global LV dysfunction (in vivo LV end-diastolic pressure, 4+/-1 versus 23+/-1.6 mm Hg; Langendorff LV developed pressure, 105+/-4 versus 62+/-9 mm Hg, P<.001 for both). Myocytes isolated from these hearts showed significant structural remodeling (LV myocytes, 24% longer and 15% wider; right ventricular myocytes, 38% longer and 31% wider, all P<.05). LV myocyte length correlated with changes in LV volume (r=.79) and function (LV developed pressure, r=-.81). However, LV myocytes from the same hearts showed normal contractile function and intracellular Ca2+ transients at baseline and during inotropic stimulation with increasing extracellular Ca2+ (1 to 6 mmol/L). The shortening-frequency relationship was also similar in myocytes from sham and myocardial infarction rats. CONCLUSIONS: Postinfarct LV remodeling occurs predominantly by myocyte lengthening rather than by myocyte slippage. However, contractile function of the unloaded myocytes from the remote noninfarcted LV myocardium of the remodeled heart is normal. Therefore, myocyte contractile abnormalities may not contribute to global dysfunction of the remodeled heart. Reduced myocyte mass and nonmyocyte factors like increased wall stress, altered LV geometry, and changes in the myocardial interstitium may be more important in the genesis of postinfarct LV dysfunction in this model. PMID- 9403623 TI - Profibrillatory effects of intracoronary thrombus in acute regional ischemia of the in situ porcine heart. AB - BACKGROUND: An intracoronary thrombus during regional ischemia is related to life threatening arrhythmias. The electrophysiological consequences of a thrombus are unknown. METHODS AND RESULTS: In open chest pigs, regional ischemia was induced by intracoronary injection of a thrombus (protocol 1). In protocol 2, coronary ligation was followed by injection of heparinized blood. Three consecutive episodes of ischemia (10 minutes) and reperfusion (20 minutes) were studied in protocols 3 and 4 (ligation). During the former, an intracoronary thrombus started the third period of ischemia. Multiple (78) local electrograms were recorded simultaneously, and activation patterns were determined. In a first period of ischemia, ventricular fibrillation (during the first 10 minutes) occurred more often after intracoronary thrombosis than during the other protocols (4/7 versus 2/19, P<.05) despite similar size of the ischemic tissue. The incidence of delayed arrhythmias (between 15 and 30 minutes) was not different. Epicardial activation delay was larger 2 to 4 minutes after intracoronary thrombosis compared with ligation. ST elevation was larger with than without a thrombus (2 minutes of ischemia, 12.9+/-4.1 versus 8.2+/-3.0 mV; +/-SD, P<.05). In protocols 3 and 4 the second period and third period of ischemia were similar irrespective of the presence of an intracoronary thrombus. CONCLUSIONS: More conduction slowing underlies the profibrillatory effect of an intracoronary thrombus relative to coronary ligation. After preconditioning with ischemia, the profibrillatory effects are no longer detectable. PMID- 9403624 TI - Nitroglycerin enhances the ability of dobutamine stress echocardiography to detect hibernating myocardium. AB - BACKGROUND: A biphasic response of wall thickening with initial improvement and subsequent deterioration during dobutamine stress echocardiography (DSE) has been increasingly used for detection of hibernating myocardium. However, the improvement of wall thickening at low-dose DSE may be limited in hibernating myocardium by severe hypoperfusion. Nitroglycerin (NTG) improves myocardial perfusion, reduces oxygen demand, and may enhance low-dose dobutamine to improve wall thickening. METHODS AND RESULTS: A pig model of myocardial hibernation of 24 hours to 7 days was created through severe left anterior descending coronary artery stenosis with coronary flow reductions of approximately 40%, producing severe regional left ventricular dysfunction but no infarction in seven pigs. Myocardial infarction was produced in five pigs with occlusion of the artery. DSE was performed with incremental doses with and without an NTG infusion of 50 to 100 microg/min. In the hibernating group, NTG alone improved wall thickening in the hibernating region modestly from 11.4+/-7.2% at baseline to 19.1+/-7.0%. The improvement was associated with increased regional coronary flow from 0.46+/-0.12 to 0.55+/-0.13 mL x beat(-1) x 100 g myocardium(-1) (P<.05). There was an additive effect of NTG to low-dose (2.5 to 5 microg x kg(-1) x min(-1)) dobutamine on wall thickening in the hibernating region. The improvement of wall thickening of hibernating myocardium with NTG and dobutamine, from 23.7+/-11.1% to 31.1+/-8.9% (P<.001), was associated with an increase in regional coronary flow (P<.01). NTG did not prevent high doses of dobutamine from inducing deterioration of wall thickening in hibernating myocardium. In the infarcted group, no improvement in wall thickening was observed in infarcted regions during NTG infusion, dobutamine infusion, or the combination. CONCLUSIONS: NTG enhances the improvement of wall thickening at low-dose dobutamine and does not prevent high-dose dobutamine from inducing ischemia in hibernating myocardium. Thus, NTG augments the biphasic response of wall thickening and improves the accuracy of DSE for detecting viable myocardium. PMID- 9403625 TI - Effect of angiotensin-converting enzyme inhibition on myocardial fibrosis and function in hypertrophied and failing myocardium from the spontaneously hypertensive rat. AB - BACKGROUND: After a period of stable hypertrophy, male spontaneously hypertensive rats (SHR) develop heart failure between 18 to 24 months of age, with depression of active myocardial function and increased passive stiffness. We tested the hypothesis that chronic ACE inhibition by captopril would prevent and possibly reverse impairment of myocardial function. METHODS AND RESULTS: Male SHR and normotensive Wistar-Kyoto rats (WKY) were assigned to no treatment or captopril treatment (2 g/L in drinking water) begun at ages 12, 18, and 21 months; animals were studied at 24 months of age, or earlier when evidence of heart failure was found in SHR (mean age, 19+/-2 months). In an additional group, captopril treatment was begun when SHR developed heart failure; surviving animals were studied at 24 months of age. In untreated SHR, relative to WKY, isometric stress development at Lmax, maximum rate of stress development, and shortening velocity were depressed, whereas passive stiffness was increased, in association with the development of myocardial fibrosis. In the SHR treated before cardiac dysfunction, captopril administration attenuated hypertrophy and prevented contractile dysfunction, fibrosis, and increased passive stiffness. Captopril treatment begun after cardiac function was impaired reduced left ventricular hypertrophy but did not restore intrinsic contractile function or reduce fibrosis or passive stiffness. CONCLUSIONS: In the male SHR, early treatment with captopril was associated with the most marked attenuation of dysfunction relative to the untreated SHR. Treatment initiated after the onset of heart failure improved clinical signs of heart failure and decreased left ventricular hypertrophy in surviving animals but did not reverse the fibrosis and contractile dysfunction associated with heart failure. PMID- 9403626 TI - Effects of quinidine on repolarization in canine epicardium, midmyocardium, and endocardium: I. In vitro study. AB - BACKGROUND: The antiarrhythmic action of quinidine is associated with a slowing of conduction and prolongation of repolarization. The latter effect has no consistent correlation with quinidine actions on action potential duration (APD) in isolated tissue experiments. To enhance our understanding of the mechanisms of quinidine action, we studied its effect on APD in canine epicardial, midmyocardial, and endocardial tissues. METHODS AND RESULTS: Standard microelectrode techniques were used to study the effects of quinidine 2.5 to 20 micromol/L on APD in ventricular epicardial, endocardial, and transmural (M-cell) slabs at cycle lengths (CLs) from 300 to 4000 ms. Qualitatively different time courses of actions and concentration- and rate-dependent effects were seen in M cells compared with the others. In endocardium and epicardium, quinidine induced monotonic and concentration-dependent APD prolongation at all CLs. In contrast, the effects of quinidine in M cells varied from prolongation to shortening, depending on duration of superfusion, concentration, and CL. Experiments with E4031 and TTX suggested that in M cells, quinidine-induced APD lengthening was attributable to block of delayed rectifier potassium current and APD shortening was due to inhibition of TTX-sensitive steady-state sodium current. CONCLUSIONS: In vitro, there is a significant difference of quinidine effects in M cells versus epicardial and endocardial cells that appears to reflect differences in the contributions of specific ion channels to the APD at the three sites. The differences may influence the actions of quinidine on repolarization of the heart in situ and determine both the proarrhythmic and antiarrhythmic actions of the drug. PMID- 9403627 TI - Effects of quinidine on repolarization in canine epicardium, midmyocardium, and endocardium: II. In vivo study. AB - BACKGROUND: In the companion article, we report a significant difference in quinidine effects on the action potential duration between surface (epicardial and endocardial) cells and midmyocardial cells (M cells) of canine left ventricle in vitro. This article considers two questions raised by the previous study: (1) Are the complex quinidine effects in vitro reflected in its actions on the heart in situ? (2) What are the cellular determinants of quinidine effects on QT interval in ECG? METHODS AND RESULTS: We used plunge and surface electrodes to measure activation-recovery intervals (ARIs) of bipolar electrograms obtained from epicardium, endocardium, and midmyocardium (3, 5, and 9 mm from epicardium) of canine left ventricle in conditions of AV block and right ventricular pacing. Quinidine was infused continuously; its plasma level increased from 1.6+/-0.1 microg/mL at 30 minutes to 7.6+/-0.7 microg/mL at 180 minutes. At cycle lengths (CLs) from 300 to 1500 ms, there was no ARI gradient across the ventricular wall before and during quinidine infusion. At a CL of 300 ms, therapeutic concentrations of quinidine prolonged ARIs and QT intervals. At a CL of 1500 ms, ARIs were significantly prolonged at low quinidine concentrations. With an increase of quinidine concentration, this effect subsided and disappeared. CONCLUSIONS: In situ, quinidine-induced prolongation of repolarization is uniform in all myocardial layers and follows the pattern observed in M cells in vitro. The ability of quinidine in therapeutic concentrations to prolong repolarization at rapid heart rates can contribute to its antiarrhythmic efficacy. PMID- 9403628 TI - Functional mechanisms underlying tachycardia-induced sustained atrial fibrillation in a chronic dog model. AB - BACKGROUND: Rapid atrial activation causes electrical remodeling that promotes atrial fibrillation (AF), but underlying mechanisms are incompletely understood. We applied epicardial mapping to evaluate atrial electrophysiology and AF duration in dogs subjected to rapid atrial pacing (400/min). METHODS AND RESULTS: Dogs paced for 1 (P1, n=7), 7 (P7, n=13), or 42 (P42, n=7) days were compared with sham dogs (P0, n=13). Atrial pacing progressively increased AF duration. Atrial effective refractory period (ERP) and ERP accommodation to rate were significantly decreased by pacing, with near-maximal changes within 7 days. Atrial conduction velocity decreased more slowly, with maximum changes at 42 days, contributing to increases in AF duration after ERP stabilized. Stepwise multilinear regression indicated that both wavelength (P=.02) and duration of pacing (P=.0001) were independent determinants of changes in AF duration. Mean atrial fibrillation cycle length (AFCL) at 112 recording sites decreased with increased duration of rapid pacing (P<.001), and the SD of AFCL increased progressively (P<.0001), together accounting for 72% of the variance in AF duration. Increases in AFCL variability were due to regionally determined differences in AFCL changes caused by rapid pacing. The number of zones of reactivation per cycle of AF increased as AF became more sustained, consistent with multiple-wavelet reentry. CONCLUSIONS: Rapid atrial activation causes time dependent decreases in ERP, conduction velocity, and wavelength, which, along with increased regional heterogeneity, provide a substrate for AF. The conduction abnormalities and increased regional heterogeneity previously noted in patients with AF may be a consequence, as well as a cause, of the tachyarrhythmia. PMID- 9403629 TI - Activation-repolarization coupling in the normal swine endocardium. AB - BACKGROUND: While abnormalities of activation and repolarization play an important role in arrhythmogenesis, little information is available on the interaction between their spatial dispersions in the heart. This study examined the effects of activation spread on the spatial distribution of the repolarization properties during different depolarization patterns. METHODS AND RESULTS: Left ventricular (LV) endocardial activation and repolarization patterns were mapped in 13 healthy pigs. LV local activation, repolarization, and activation-recovery interval (ARI) times were determined from the intracardiac unipolar electrograms, color-coded, and superimposed on a three-dimensional anatomic map of the ventricle generated with a nonfluoroscopic mapping system. ARI values correlated with the duration of monophasic activation potential recorded from onset of activation to time of 90% repolarization (r=.97, P<.01). Activation time range of the left ventricle was 42+/-5 ms (mean+/-SEM) during sinus rhythm and 54+/-5 ms during right ventricular septal pacing. ARI inversely correlated with the corresponding activation times during both sinus (r2=.76+/ .03) and paced (r2=.77+/-.02) rhythms. The longest ARIs were located at the sites of earliest activation and shortest at the latest activation areas, with gradual shortening between them. CONCLUSIONS: The spatial distribution of repolarization is dependent on the activation pattern. Repolarization dispersion in the healthy swine heart is relatively small as the result of tight coupling of the action potential duration to the activation process, assigning longer ARIs to sites activated earlier. This coupling reduces global and regional dispersion of repolarization and may serve as an important antiarrhythmic mechanism present in normal myocardium. PMID- 9403631 TI - Electrophysiological effects of acute dilatation in the isolated rabbit heart: cycle length-dependent effects on ventricular refractoriness and conduction velocity. AB - BACKGROUND: Acute ventricular dilatation has important electrophysiological effects: Dilatation shortens action potential duration and refractoriness without an apparent effect on conduction velocity. These effects have been implicated as a potential mechanism of arrhythmias in patients with congestive failure. Because the influence of cycle length on these phenomena has not been studied, we examined the effects of dilatation during ventricular pacing at cycle lengths from 1000 to 150 ms. METHODS AND RESULTS: Thin epicardial layers were created in isolated, perfused rabbit left ventricles (n=7). A fluid filled latex balloon was secured in the left ventricle to dilate the left ventricle. Mapping was performed with 248 epicardial electrodes. Longitudinal conduction velocity (76+/-1 cm/s; mean+/-SEM) and transverse conduction velocity (26+/-1 cm/s) were not influenced by dilatation at any cycle length. In contrast, the effects of dilatation in decreasing left ventricular effective refractory period (ERP) were significantly greater at shorter drive cycle lengths: The decrease in ERP was 2+/-2 ms (a 1% change) at a drive cycle length of 1000 ms and 18+/-4 ms (a 20% change) at a drive cycle length of 150 ms. In 10 additional intact, isolated perfused rabbit hearts, dilatation decreased ERP to a greater degree during 250 ms drive cycle length pacing than during pacing at 400 ms (25+/-4 versus 16+/-3 ms; P=.01). CONCLUSIONS: Acute dilatation exaggerates the normal rate-dependent shortening of refractoriness but does not influence transverse or longitudinal conduction velocity. This observation suggests that the electrophysiological effects of acute dilatation may be greater during tachycardia than at slower cycle lengths. This may have implications for arrhythmias in patients with congestive heart failure. PMID- 9403630 TI - Intrapericardial delivery of L-arginine reduces the increased severity of ventricular arrhythmias during sympathetic stimulation in dogs with acute coronary occlusion: nitric oxide modulates sympathetic effects on ventricular electrophysiological properties. AB - BACKGROUND: Nitric oxide (NO) modulates autonomic effects on myocardial contractility and sinus and atrioventricular nodal function of the heart. Whether NO influences autonomic actions on ventricular electrophysiological properties and arrhythmogenesis is not known. METHODS AND RESULTS: Four groups consisting of 43 autonomically denervated dogs were studied. To "superfuse" sympathetic nerves innervating the ventricles, test drugs were introduced into the pericardial sac for 30 minutes, and their effects on ventricular effective refractory period (ERP) and arrhythmia development were assessed before and during sympathetic stimulation (SS). In group 1 (n=12), ventricular ERPs showed no significant difference between control and superfusion with L-arginine, a NO precursor (222+/ 20 versus 222+/-19 ms, P=.485). However, L-arginine significantly reduced SS induced ERP shortening compared with control (9+/-7 versus 13+/-7 ms, P<.001). Simultaneous administration of N(G)-monomethyl-L-arginine (2 mg/mL) abolished the inhibitory effects of L-arginine (13+/-7 versus 13+/-7 ms, P=.885). In group 2 (n=15), the severity of ventricular arrhythmias significantly increased during SS. L-Arginine reduced this increase caused by SS. In group 3 (n=8), plasma norepinephrine spillover measured from the coronary sinus significantly increased during SS and was reduced by pericardial superfusion with L-arginine compared with control (6005.2+/-1525.6 versus 8503.4+/-2044.5 pg/min, P=.012). In group 4 (n=8), L-arginine pericardial superfusion significantly increased NO overflow measured from the coronary sinus during SS (93.25+/-59.20 versus 114.82+/-74.92 nmol/min, P=.043). CONCLUSIONS: Pericardial L-arginine reduces ERP shortening and increased severity of ischemic ventricular arrhythmias during SS in dogs. NO induced reduction of norepinephrine release in the heart may be one of the underlying mechanisms. PMID- 9403632 TI - Nonuniform heating during radiofrequency catheter ablation with long electrodes: monitoring the edge effect. AB - BACKGROUND: Long, narrow electrodes are being considered for radiofrequency ablation of atrial fibrillation; however, preliminary work revealed coagulum formation on the electrodes and lack of lesion continuity. This may be due to the "edge effect," which concentrates radiated energy at sharp geometric gradients. It is proposed that temperature sensors at electrode edges are preferable to a single centered sensor for temperature feedback and monitoring of long electrode geometries. METHODS AND RESULTS: A finite element model was used to predict the heating properties of new long electrode geometries. Sixteen dogs with atrial fibrillation underwent left and right atrial ablation using catheters with multiple 12.5-mm coil electrodes. Electrodes with a single thermistor were compared with electrodes with dual thermocouples placed at opposite ends and on opposing sides of the electrode. Power, temperature, and impedance were recorded for all lesions, and coagulum adhesion and magnitude were noted in a subset of lesions. Finite element analysis shows uneven heating, with the main heating concentrated at the electrode edges and a propensity toward temperatures >100 degrees C with single-thermistor feedback control. Ablations with dual thermocouple electrodes achieved higher measured temperatures at lower power levels than those that used single-thermistor electrodes. Impedance rises and coagulum adherence occurred less frequently with dual thermocouple electrodes than with single, centered thermistor electrodes (176 of 395 versus 9 of 425 lesions; P<.0001; 46 of 98 versus 7 of 150 lesions; P<.0001, respectively). CONCLUSIONS: Maximum heating from radiofrequency energy occurs at the electrode edges, particularly with long electrodes. The safety of temperature-feedback atrial ablation with these electrodes is significantly improved by monitoring temperatures at the edges. PMID- 9403633 TI - Extracellular matrix remodeling in heart failure: a role for de novo angiotensin II generation. PMID- 9403635 TI - Roles of infectious agents in atherosclerosis and restenosis: an assessment of the evidence and need for future research. PMID- 9403634 TI - Modulation of growth factor action: implications for the treatment of cardiovascular diseases. AB - Peptide growth factors are involved in fundamental cellular processes relevant for cardiovascular physiology and pathology, namely, atherogenesis and angiogenesis. The modulation of growth factor-related signals represents a novel strategy for the treatment of cardiac and vascular disease. Experimental modulation of growth factor action has already provided a better understanding of cardiovascular biology and pathophysiology. In turn, the development of specific and powerful molecular tools is setting the stage for the exploration of their clinical potentials. Current strategies include the use of recombinant proteins, specific inhibitors of protein-protein interactions, tyrosine kinase inhibitors, the generation and application of dominant-negative molecules, the development of antisense strategies, and a variety of different gene transfer approaches. Parallel avenues of research are heading toward the same goal, the specific suppression of potent pathogenic stimuli that induce and promote atherogenesis or the augmentation of beneficial ones such as induction of therapeutic angiogenesis. The successful application of one of these strategies seems to be in reach and will certainly be a milestone in molecular medicine. PMID- 9403636 TI - Insulin as a vascular and sympathoexcitatory hormone: implications for blood pressure regulation, insulin sensitivity, and cardiovascular morbidity. AB - The past several years have witnessed a major surge of interest in the cardiovascular actions of insulin. This interest has stemmed on the one hand from epidemiological studies that demonstrated an association between obesity, insulin resistance, and hypertension, leading to the so-called insulin hypothesis of hypertension. On the other hand, this interest has been stimulated by experimental evidence suggesting that the vascular actions of insulin may play a role in its main action, namely the promotion of glucose uptake in skeletal muscle tissue. Two tenets have emerged about how insulin may exert its cardiovascular actions. First, it is now firmly established that acute insulin administration stimulates sympathetic nerve activity in both animals and humans. Second, there is increasing evidence that insulin stimulates muscle blood flow, an effect that appears to be mediated at least in part by an endothelium dependent mechanism. This review summarizes the current understanding and gaps in knowledge on cardiovascular actions of insulin in humans and pathophysiological consequences of derangements of such actions. PMID- 9403637 TI - Images in cardiovascular medicine. Effect of hiccuping on atrial septum. PMID- 9403638 TI - Images in cardiovascular medicine. Three-dimensional computed tomography of corrected transposition of the great arteries. PMID- 9403639 TI - Recombinant erythropoietin for the anemia of prematurity: still a promise, not a panacea. PMID- 9403640 TI - Are certain children more likely to develop neuroblastoma? PMID- 9403641 TI - Beyond hydrolysates: use of L-amino acid formula in resistant dietary protein induced intestinal disease in infants. PMID- 9403642 TI - The effect of erythropoietin on the transfusion requirements of preterm infants weighing 750 grams or less: a randomized, double-blind, placebo-controlled study. AB - BACKGROUND: Clinical trials of erythropoietin (EPO) administration to preterm infants have not focused on infants weighing 750 gm or less, the population most likely to receive multiple transfusions because of large phlebotomy losses. It is unknown whether preterm infants weighing 750 gm or less will respond to EPO by accelerating erythropoiesis, or whether EPO administered to this population will decrease blood transfusions. METHODS: We randomly assigned 28 extremely low birth weight preterm infants (mean +/- SEM: 24.7 +/- 0.3 weeks' gestation, 662 +/- 14 gm birth weight), in the first 72 hours of life, to receive either EPO (200 U/kg/day) or placebo for 14 days and administered transfusions only according to protocol over a 21-day study period. All infants received 1 mg/kg/day iron dextran in their total parenteral nutrition solution during the 14-day treatment period. RESULTS: During the 21-day study period, a lower number and volume of transfusions were received by the EPO recipients (4.7 +/- 0.7 transfusions per patient and 70 +/- 11 ml/kg per patient) than by the placebo recipients (7.5 +/- 1.1 transfusions per patient and 112 +/- 17 ml/kg per patient; p < 0.05, EPO vs placebo), whereas hematocrits remained similar in the two groups. Reticulocyte counts were similar in both groups on day 1 but were greater in the EPO recipients on day 14 (EPO day 1, 351 +/- 53; EPO day 14, 359 +/- 40 x 10(3)/microl; placebo day 1, 334 +/- 64; placebo day 14, 120 +/- 10 x 10(3)/microl; p < 0.01, EPO vs placebo). Serum ferritin concentrations were similar in both groups at the beginning of the study but were greater in the placebo recipients by day 14 (EPO, 262 +/- 44 microg/L; placebo, 593 +/- 92 microg/L; p < 0.01). No adverse effects of EPO or iron were noted. CONCLUSION: The combination of EPO and parenteral iron stimulates erythropoiesis in preterm infants weighing 750 gm or less and results in fewer transfusions during their first 3 weeks of life. PMID- 9403643 TI - Neuroblastoma and related tumors in Turner's syndrome. AB - OBJECTIVES: The identification of constitutional cytogenetic abnormalities in patients with cancer may indicate loci of genes, abnormalities of which are responsible for tumor development or progression. This study was undertaken to determine whether girls with Turner's syndrome (TS) (partial or complete deletion of an X chromosome, short stature, gonadal dysgenesis) are at increased risk of neural crest-derived tumors. STUDY DESIGN: Medical records of 394 patients with TS who were followed up at Thomas Jefferson Hospital and Children's Hospital of Pittsburgh were reviewed for documentation of TS phenotype, constitutional cytogenetics, and history of neuroblastoma or related tumors. Informative cases were reviewed for tumor pathology, primary site, disease stage, associated symptoms, treatment, and outcome. RESULTS: Three patients were found to have neuroblastoma. A fourth child who died of neurofibrosarcoma was found to have extensive areas of ganglioneuroma, the benign counterpart of neuroblastoma, at autopsy. An additional four girls with TS and neuroblastoma were identified in the literature, as were two more patients with ganglioneuroma. These 10 patients ranged in age from 1 week to 16 10/12 years (median age, 3 years), and all but two of the children had localized lesions. Two of the seven children with neuroblastoma had courses complicated by opsoclonus-myoclonus, a syndrome found in fewer than 5% of all patients with neuroblastoma. CONCLUSIONS: These data strongly suggest that girls with TS are predisposed to the development of neuroblastoma and related tumors. Because these tumors are often of limited stage and may be underdiagnosed, screening of urine of patients with TS for elevated catecholamine metabolite levels may strengthen this association. PMID- 9403644 TI - High birth weight and risk of specific childhood cancers: a report from the Children's Cancer Group. AB - OBJECTIVES: High birth weight has been associated with a number of childhood cancers. This study was conducted to test the hypothesis that elevated birth weight is associated with an increased risk of diagnosis-specific and age specific groups of childhood cancers. METHODS: A case-control study, using a large Children's Cancer Group database, examined birth weight as a risk factor for childhood cancer. Birth weight information for the index child was available for 3711 cases and 816 control subjects. RESULTS: There was a statistically significant increased risk of acute lymphoblastic leukemia, Wilms' tumor, and neuroblastoma with increasing birth weight (p, trend = 0.006, 0.003, and 0.001, respectively). A statistically significant decreased risk of cancer was observed for soft tissue sarcoma (p, trend = 0.04). When data were stratified on the basis of age at diagnosis, many of these associations were apparent for children whose disease was diagnosed before the age of 2 years. Moreover, for acute myeloid leukemia, age at diagnosis was an important effect modifier. For children with acute myeloid leukemia whose disease was diagnosed before 2 years of age, there was a statistically significant increased risk with high birth weight (odds ratio = 2.5, 95% confidence interval 1.1 to 5.5); there was no increased risk of acute myeloid leukemia with high birth weight noted for children whose disease was diagnosed after 2 years of age (odds ratio 1.3, 95% confidence interval 0.8 to 2.2). CONCLUSIONS: Biologic studies are needed to address why high birth weight may increase risk (particularly at younger ages) of development of certain cancers. PMID- 9403645 TI - Prognostic signs in the surgical management of plexiform neurofibroma: the Children's Hospital of Philadelphia experience, 1974-1994. AB - OBJECTIVES: To estimate the rate of progression of plexiform neurofibroma after surgery and to identify prognostic factors that predict progression. STUDY DESIGN: A retrospective review of the inpatient and outpatient records of 121 patients, who had 302 procedures on 168 tumors over a 20-year period at a single large pediatric referral center. Data on age, location, indication for surgery, and extent of resection was analyzed for prognostic significance. RESULTS: The overall freedom from progression was 54%. Children < 10 years old had a shorter interval of tumor control than older children (p = 0.0004). Tumors of the head/neck/face fared worse than tumors of the extremities (p = 0.0003). Less extensive resection predicted shorter interval to progression (p < 0.0001). Indication for surgery was not of prognostic importance. In multivariable analysis older age and location in the extremities were predictors of a better outcome. CONCLUSIONS: Tumor progression is a serious problem for children with plexiform neurofibroma. Younger children, children with tumors of the head/neck/face, and tumors that cannot be nearly completely removed are at particular risk. These data may be useful in helping clinicians decide which patients and which tumors are most likely to benefit from surgical intervention. PMID- 9403646 TI - Equivalent antipyretic activity of ibuprofen and paracetamol in febrile children. AB - The antipyretic activity of ibuprofen in the Sparklets form was compared, in an equivalence study, with that of paracetamol in the same formulation. The study was conducted as a double-blind multicenter trial, with random allocation of the treatments. One hundred sixteen children of both sexes, aged 4.1 +/- 2.6 years, with a fever related to an infectious disease and a mean temperature of 39 degrees +/- 0.5 degrees C at the time of inclusion, were treated with single doses of either 10.3 +/- 1.9 mg/kg of ibuprofen or 9.8 +/- 1.9 mg/kg of paracetamol. The subjects' rectal temperature was regularly monitored for 6 hours. The statistical analysis of the results confirmed that ibuprofen and paracetamol are equivalent with respect to the following criteria (1) time elapsed between dosing and the lowest temperature: 3.61 +/- 1.34 hours for ibuprofen and 3.65 +/- 1.47 hours for paracetamol (95% confidence interval [CI] of the difference: -0.48; +0.56); (2) extent of the temperature decrease: 1.65 degrees C +/- 0.80 degrees C for ibuprofen and 1.50 degrees C +/- 0.61 degrees C for paracetamol, (95% CI of the difference: -0.41; +0.11); (3) rate of temperature decrease: 0.52 +/- 0.32 degrees C/hr for ibuprofen and 0.51 degrees C +/- 0.38 degrees C/hr for paracetamol (95% CI of the difference: -0.45; +0.55); (4) duration of temperature below 38.5 degrees C: 3.79 +/- 1.33 hours for ibuprofen and 3.84 +/- 1.22 hours for paracetamol (95% CI of the difference: 0.14; +0.12). PMID- 9403647 TI - Response to the varicella vaccine in children with nephrotic syndrome. AB - Varicella vaccine was administered to seven children with corticosteroid sensitive nephrotic syndrome. Immunization was not associated with any significant reactions or with increased frequency of relapse. The antibody response was, however, variable and a second dose was necessary before seroconversion was achieved in four patients. The findings indicate that immunization with varicella vaccine is safe in children with nephrotic syndrome in remission, but that a two-dose vaccine schedule should be considered. PMID- 9403648 TI - Enteral glutamine supplementation for very low birth weight infants decreases morbidity. AB - Glutamine, described as a "conditionally essential" amino acid for critically ill patients, has not been routinely added to parenteral amino acid formulations for critically ill neonates and is provided in only small quantities by the enteral route when enteral intake is low. We conducted a blinded, randomized study of enteral glutamine supplementation in 68 very low birth weight neonates randomly assigned to receive glutamine-supplemented premature formula versus premature formula alone between days 3 and 30 of life. Primary end points consisted of hospital-acquired sepsis, tolerance to subsequent enteral feedings (days with no oral intake), and duration of hospital stay. Hospital acquired sepsis was 30% (control group) and 11% (glutamine group). Logistic regression with birth weight as a covariate showed that: (1) feeding group was significant (p = 0.048) in determining the probability of developing proven sepsis over the course of hospitalization and (2) the estimated odds of developing sepsis were 3.8 times higher for infants in the control group than for those treated with glutamine. Glutamine-supplemented infants had better tolerance to enteral feedings as measured by percent of days on which feedings needed to be withheld (mean percentage of 8.8 vs 23.8, p = 0.007). Analysis of T cells demonstrated a blunting of the rise in HLA-DR+ and CD16 subsets in glutamine-supplemented infants. There were no differences in growth; in serum ammonia, urea, liver transaminase, or prealbumin concentrations; or in mean hospital stay. This study provides evidence for decreased morbidity in very-low-birth-weight neonates who receive enteral glutamine supplementation. PMID- 9403649 TI - Essential fatty acid status in children with cholestasis, in relation to serum bilirubin concentration. AB - The liver plays a central role in the metabolism of polyunsaturated fatty acids. We studied the relationship between essential fatty acid (EFA) status and indicators of liver function in 15 children with chronic cholestasis aged 9 months to 3.4 years (median, 1.3 years). Compared with 13 control children, the patients studied had low percentage values of phospholipid EFAs, particularly of the omega-6 fatty acids linoleic acid (18:2omega-6) and arachidonic acid (20:4omega-6). Fatty acid values exhibited an inverse relationship to serum bile acids, as well as to serum bilirubin. Bilirubin values were unrelated to the EFA precursors linoleic acid and alpha-linolenic acid but correlated inversely with the long-chain metabolites arachidonic acid (r = -0.75; p = 0.001), docosapentaenoic acid (22:5omega-3; r = -0.63; p = 0.01), and docosahexaenoic acid (22:6omega-3; r = 0.72; p = 0.002). We conclude that children with chronic cholestasis are at a high risk for EFA deficiency, which increases with progressive elevation of serum bilirubin. Hepatic conversion of essential precursor fatty acids into their long-chain metabolites may be increasingly impaired with advancing severity of liver disease. PMID- 9403650 TI - Linear growth characteristics of children with cleft lip and palate. AB - OBJECTIVE: To study the linear growth characteristics of children with isolated cleft lip (CL), cleft palate (CP), or both (CLP) and to determine whether this population is at risk for short stature. STUDY DESIGN: Retrospective chart review identified 324 patients with CL, CP, or CLP that displayed no additional congenital anomalies. Longitudinal height and growth rate analyses were performed on routine anthropometric measurements gathered from hospital and clinic records. One-sample t tests (p < 0.05) of average height percentiles were performed at yearly intervals. Analysis of variance was performed on clefting subgroups. RESULTS: From birth to 10 years of age, the average height of both male and female white patients is consistently near the 40th percentile. At yearly intervals, 60% of male and 70% of female average heights demonstrate statistical difference from the population mean. For all patients, 64% of male but only 36% of female growth rates, from 2.5 to 12 years of age, were above the population mean. CONCLUSIONS: White children from birth to 10 years of age with isolated CL, CP, or CLP demonstrated a mean height below the population mean. These data suggest that children with isolated clefting manifest an intrinsic tendency toward short stature. In addition, male patients display above-average growth rates, whereas female patients display below-average growth rates, from 2 to 18 years of age. The data imply that female patients may be at increased risk of overall short stature, whereas male patients may eventually obtain mean population height. PMID- 9403651 TI - Prognosis for survival and improvement in function in children with severe developmental disabilities. AB - OBJECTIVE: To derive prognostic data for survival and clinical improvement in children with severe developmental disabilities. STUDY DESIGN: A 13-year follow up study of several cohorts of children initially evaluated before their first birthday. The outcomes studied were survival and improvement in condition. Methods were used to overcome limitations in previously published work on the same California data base. Of the 11,912 children who received services from the California Department of Developmental Services between January 1980 and December 1993, we focused on three cohorts defined according to mobility and need for tube feeding. RESULTS: Children who were tube fed and unable to lift their heads by ages 3 to 12 months were at high risk for early death, with a median remaining life expectancy of 3.2 years. Of those who survived an additional 2 years, the condition of about one third improved. A substantial majority of those who either showed improvement or died had done so by that age. CONCLUSION: By age 5 years, the prognoses for survival and improvement have to a large extent been clarified. For children who survive to age 5 years, even those in the lowest functioning cohort have a 60% chance of surviving an additional 5 years. Detailing the probabilities of various outcomes at various ages should be useful to parents, pediatricians, and others concerned with children with developmental disabilities. PMID- 9403652 TI - Heavy prenatal alcohol exposure with or without physical features of fetal alcohol syndrome leads to IQ deficits. AB - OBJECTIVE: To assess general intellectual functioning in children with histories of heavy prenatal alcohol exposure, with or without the facial features and growth deficiencies characteristic of fetal alcohol syndrome (FAS). DESIGN: Forty seven alcohol-exposed children were recruited on evaluation at a dysmorphology clinic and evaluated as part of a university research project using standard tests of IQ. Thirty-four of the alcohol-exposed patients met the traditional diagnostic criteria for FAS. The other 13 alcohol-exposed children lacked both the pattern of facial features and prenatal or postnatal growth deficiency characteristic of the diagnosis. RESULTS: Compared with normal control subjects matched for age, sex, and ethnicity, both groups of alcohol-exposed children displayed significant deficits in overall IQ measures and deficits on most of the subtest scores. Although those in the nondysmorphic group usually obtained marginally higher IQ scores than those in the FAS group, few significant differences were found between the two alcohol-exposed groups. CONCLUSIONS: These results indicate that high levels of prenatal alcohol exposure are related to an increased risk for deficits in intellectual functioning and that these can occur in children without all of the physical features required for a diagnosis of FAS. They also emphasize the need for conducting a thorough history of prenatal alcohol exposure in children with intellectual deficits. PMID- 9403653 TI - Suppression and recovery of the neonatal hypothalamic-pituitary-adrenal axis after prolonged dexamethasone therapy. AB - OBJECTIVE: To evaluate the duration and level of hypothalamic-pituitary-adrenal (HPA) axis suppression in premature infants treated with a prolonged course of glucocorticoids for chronic lung disease. STUDY DESIGN: We evaluated HPA axis function in nine very low birth weight (VLBW) infants before and 48 hours after a prolonged (14 to 42 days) dexamethasone (Dex) course. Seven of these infants underwent serial testing in the Clinical Research Center to evaluate the time course of HPA axis recovery. Adrenal function was assessed directly with synthetic adrenocorticotropic hormone (ACTH) stimulation, pituitary function with ovine corticotrophin releasing hormone (oCRH) stimulation, and combined axis function with 3-hour metyrapone testing. RESULTS: Baseline cortisol values were higher before Dex therapy (18.6 +/- 3.9 microg/dl; mean +/- SEM) than after (5.77 +/- 1.45 microg/dl; p < 0.01), as were ACTH-stimulated cortisol levels (24.8 +/- 1.7 microg/dl vs 12.0 +/- 2.2 microg/dl; p < 0.001). ACTH response to oCRH decreased after Dex treatment (22.8 +/- 7.6 pg/ml vs 11.5 +/- pg/ml), but this was not statistically significant (p = 0.18). 11-Deoxycortisol (11-DOC) response to metyrapone dropped from 11.1 +/- 0.5 microg/dl to 4.7 +/- 1.0 microg/dl after Dex therapy (p < 0.0001). Longitudinal testing reveals that adrenal suppression may be short-lived, while recovery of higher centers is more delayed. CONCLUSIONS: Basal cortisol levels may be used as a screening test, but if the level is less than 15 microg/dl, more definitive testing should be performed. The sluggish recovery of higher HPA axis centers is most reliably evaluated by using 11-DOC response to a single dose of metyrapone in VLBW infants after prolonged Dex therapy. PMID- 9403654 TI - Consequences of irregular versus continuous medical follow-up in children and adolescents with insulin-dependent diabetes mellitus. AB - OBJECTIVES: To study the social and family characteristics of patients with insulin-dependent diabetes mellitus with irregular versus continuous clinical follow-up and to study the medical outcomes of patients with these follow-up patterns. METHODS: An onset cohort of 61 children and adolescents with insulin dependent diabetes mellitus and their parents were studied. Aspects of their social and family environment were assessed at study inception and examined in relation to frequency of follow-up early in the course of the illness. Follow-up was dichotomized so that patients with continuous follow-up were compared with patients with irregular follow-up, who were defined as those missing 1 full year of planned medical appointments during the second through fourth years after diagnosis. Patients with irregular and continuous follow-up were compared in terms of acute metabolic complications, glycemic control, and retinopathy status during a 10-year period. RESULTS: Compared with individuals with continuous follow-up, patients with irregular clinical visits were more likely to be from families of lower socioeconomic class levels, have a parental history of separation and divorce, and were members of families that reported being least openly expressive of positive emotions. Poor glycemic control in year 1 was associated with irregular follow-up in years 2 through 4. Patients with irregular follow-up continued to have worse glycemic control in years 2 through 4 than patients with continuous follow-up. However, in years 7 and 10 their glycemic control no longer differed from patients with continuous follow-up. More episodes of diabetic ketoacidosis occurred in the irregular follow-up group. Finally, retinopathy occurred more frequently among those in the irregular follow-up group. CONCLUSION: Early irregular clinical follow-up should be considered a risk factor for complications of insulin-dependent diabetes mellitus. PMID- 9403655 TI - Ultrasonography of the optic nerves: clinical application in children with pseudotumor cerebri. AB - OBJECTIVE: Pseudotumor cerebri (PTC) in children has a wide spectrum of clinical presentations, from headache, which may be posterior and associated with nausea, vomiting or diplopia, to nonspecific headache, which may not be posterior and related or unrelated to other symptoms. In children with nonspecific headache, supportive evidence for diagnosis may be required before invasive procedures such as lumbar punctures are performed. Ultrasonography of the optic nerves (USON) is a noninvasive examination that can provide information about optic nerve changes, including those resulting from increased intracranial pressure. The applicability of USON in the diagnosis and follow-up of PTC was examined. STUDY DESIGN: Seventeen children were referred to our service because of a clinical suspicion of PTC, suggested by the presence of swollen optic nerve discs and/or headache, without clinical evidence of another cause. All had normal brain computed tomography and/or magnetic resonance imaging results before referral. Each child was examined with USON while in the supine position and with a 30-degree head tilt and underwent a lumbar puncture. USON was repeated on follow-up evaluation. RESULTS: The diagnosis of PTC was confirmed by lumbar puncture in 10 children and ruled out in 6 children. Overall, the USON results correlated with an increased opening pressure on lumbar puncture in 11 children. CONCLUSION: We noted an excellent correlation between the clinical results and the USON findings in PTC, and in many cases repeated lumbar punctures could be avoided. USON is more easily applied than a lumbar puncture, without the accompanying risks. It may be used as an indicator of increased intracranial pressure, as well as a follow-up tool. However, further studies are required before the accuracy of USON can be fully established. PMID- 9403656 TI - Intolerance to protein hydrolysate infant formulas: an underrecognized cause of gastrointestinal symptoms in infants. AB - The purpose of this study was to determine the effectiveness of an amino acid based infant formula in infants with continued symptoms suggestive of formula protein intolerance while they were receiving casein hydrolysate formula (CHF). Twenty-eight infants, 22 to 173 days of age, were enrolled; each had received CHF for an average of 40 days (10 to 173 days) and continued to have bloody stools, vomiting, diarrhea, irritability, or failure to gain weight, or a combination of these symptoms. Sigmoidoscopy with rectal biopsy was performed in all infants. The infants then received an amino acid-based infant formula, Neocate, for 2 weeks. After 2 weeks of treatment, 25 of the infants demonstrated resolution of their symptoms and underwent challenge with CHF. Of the 25 who were challenged, eight tolerated the CHF and the remainder had recurrence of their symptoms. The histologic features in these infants varied from eosinophilic infiltration to normal. We conclude that not all infants with apparent formula protein-induced colitis respond to CHF and that these infants may have resolution of their symptoms when fed an amino acid-based infant formula. PMID- 9403658 TI - Effects of milk-borne colony stimulating factor-1 on circulating growth factor levels in the newborn infant. AB - Colony stimulating factor-1 (CSF-1) concentrations in colostrum were 20 to 25 times higher than in serum at birth and declined with lactation. No difference in concentrations of circulating CSF-1, however, were noted between breast-fed and formula-fed infants, suggesting that milk-borne CSF-1 may feed back negatively on endogenous growth factor levels, may act locally in the gastrointestinal tract, or may be locally degraded. PMID- 9403657 TI - Allergy to extensively hydrolyzed cow milk proteins in infants: identification and treatment with an amino acid-based formula. AB - We report on 13 infants allergic to extensively hydrolyzed protein formulas (EHFs) who were first seen with chronic digestive symptoms. Feeding with an amino acid-based formula decreased symptoms and improved weight gain. A challenge with EHF produced positive results in all these infants. Allergy to EHF must be considered in patients who are allergic to cow milk and have persisting symptoms on an EHF diet. PMID- 9403659 TI - Postviral gastroparesis: presentation, treatment, and outcome. AB - We describe the clinical features and long-term outcome of 11 children who had persistent gastroparesis after an acute viral illness, eight of whom tested positive for rotavirus. Gastric emptying was delayed in the 10 children evaluated with scintigraphy. Antroduodenal manometry confirmed postprandial antral hypomotility in 10 subjects. All children recovered within 6 to 24 months. PMID- 9403660 TI - Computer-assisted adjustment of inspired oxygen concentration improves control of oxygen saturation in newborn infants requiring mechanical ventilation. AB - Open-loop computer control of inspired oxygen concentration was evaluated in 16 newborn infants requiring mechanical ventilation. FIO2 and oxygen saturation were compared for 2-hour periods of computer versus routine manual FIO2 adjustment. During computer-assisted FIO2 adjustment, patients spent more time at the target SaO2 and less time with SaO2 < 90%. PMID- 9403661 TI - Discontinuation of long-term transfusion therapy in patients with sickle cell disease and stroke. AB - Long-term transfusion therapy in patients with sickle cell disease and stroke markedly decreases the risk of stroke recurrence. However, it is not known how long the transfusions should be continued. Published reports have documented a high risk of stroke recurrence after stopping transfusion. We report on nine consecutive patients with sickle cell disease and stroke whose long-term transfusion therapy was discontinued and in whom no ischemic strokes developed during 80.75 patient years of follow-up. PMID- 9403662 TI - Umbilical venous catheterization and the risk of portal vein thrombosis. AB - Portal vein thrombosis has been associated with umbilical venous catheterization. We studied the incidence of portal vein thrombosis associated with umbilical venous catheterization with the catheter tip not in the portal venous system. Appropriate placement of an umbilical venous catheter in sick neonates is associated with a low risk of portal vein thrombosis (actual incidence, 1.3%). PMID- 9403663 TI - Development of systemic to pulmonary collateral arteries in premature infants. AB - Infants with congenital heart disease and chronic lung disease are at risk for development of systemic-to-pulmonary collateral arteries (SPCA). This study characterizes associated clinical findings in 20 premature infants without CHD who were diagnosed as having SPCA with echocardiography. SPCA can occur in premature infants without chronic lung disease and may represent a transient phenomenon. PMID- 9403664 TI - Hypoparathyroidism in a patient with long-chain 3-hydroxyacyl-coenzyme A dehydrogenase deficiency caused by the G1528C mutation. AB - Mitochondrial trifunctional protein (MTP), an enzyme complex participating in fatty acid beta-oxidation, is the potential site of two documented defects: long chain 3-hydroxyacyl-coenzyme A dehydrogenase (LCHAD) and MTP deficiencies. LCHAD deficiency usually manifests as hypoglycemia, with hepatopathy, hypotonia, cardiomyopathy, and retinopathy. Hypoparathyroidism has been detected in a patient with MTP deficiency. We now report on a patient with LCHAD deficiency and hypoparathyroidism, evidenced by hypocalcemia, hyperphosphatemia, and a low level of parathyroid hormone, in whom the parathyroid glands could not be located after death. PMID- 9403665 TI - Overflow encopresis and stool toileting refusal during toilet training: a prospective study on the effect of therapeutic efficacy. AB - We determined the incidence of stool toileting refusal in 53 children with overflow encopresis; 24 (45%) experienced difficulty toilet training for bowel movements. One-year follow-up data were obtained for 43 children, 31 with secondary encopresis and 12 with primary encopresis. Among the children with secondary encopresis, no difference was observed in response to treatment between children with and without difficulty toilet training. Ninety-one percent (11 of 12) of the children who had been soiling for less than 1 year at the time of presentation were free of soiling and no longer required therapeutic medication, compared with 55% (10 of 18) of the children who had been soiling for a longer period. Only 1 of 12 children with primary encopresis was free of soiling and no longer receiving therapeutic medication at 1 year, compared with 21 of 37 with secondary encopresis (p = 0.003). We concluded that children with primary encopresis who demonstrated stool toileting refusal during toilet training were resistant to medical treatment. PMID- 9403666 TI - Osteopenia in Rett syndrome. AB - Bone density analysis, dietary intake, and anthropometrics were compared in 20 subjects with Rett syndrome (RS), 25 normal control subjects, and 11 girls with cerebral palsy. Bone mineral density, bone mineral content, and spine (bone) mineral density were significantly reduced in the RS group. When weight and age were kept constant, the bone density was still reduced in the patients with RS. Subjects with RS are at risk for osteoporosis. PMID- 9403667 TI - Tietze's syndrome in children and infants. AB - Tietze's syndrome, characterized by isolated swelling and tenderness of a costochondral junction, usually occurs in adults. We describe eight cases of Tietze's syndrome in children, four of them aged 1 year or less. The clinical aspects and laboratory and imaging findings should enable the clinician to recognize this benign entity, thereby avoiding invasive diagnostic procedures to rule out other conditions. PMID- 9403668 TI - Lymphadenopathy and pulmonary infiltrates in a 12-year-old girl. PMID- 9403669 TI - The central issue in precocious puberty. PMID- 9403670 TI - Neurofibromatosis type I and precocious puberty: beyond the chasm. PMID- 9403671 TI - Seizures caused by chloral hydrate sedative doses. PMID- 9403672 TI - $40m plant genome sequencing effort targets the best science. PMID- 9403673 TI - UK accused over risk of CJD in plasma...and latest BSE beef ban 'comes too late'. PMID- 9403674 TI - Asia/Pacific 'bionetwork' comes to life. PMID- 9403675 TI - How to evaluate journal impact factors. PMID- 9403676 TI - Standards for safety cabinets. PMID- 9403677 TI - Learning and memory. Never fear, LTP is hear. PMID- 9403678 TI - Genome sequencing. New tricks of tick-borne pathogen. PMID- 9403680 TI - Deafness. Sounds from the cochlea. PMID- 9403679 TI - The painless synergism of aspirin and opium. PMID- 9403681 TI - Anticancer drugs. Ringing the changes. PMID- 9403682 TI - An E2F-like repressor of transcription. PMID- 9403683 TI - Ribonuclease inhibits Kaposi's sarcoma. PMID- 9403684 TI - Mosaicism in Turner's syndrome. PMID- 9403686 TI - Emergence of symbiosis in peptide self-replication through a hypercyclic network. AB - Symbiosis is an association between different organisms that leads to a reciprocal enhancement of their ability to survive. Similar mutually beneficial relationships can operate at the molecular level in the form of a hypercycle, a collective of two or more self-replicating species interlinked through a cyclic catalytic network. The superposition of cross-catalysis onto autocatalytic replication integrates the members of the hypercycle into a single system that reproduces through a second-order (or higher) form of nonlinear autocatalysis. The hypercycle population as a whole is therefore able to compete more efficiently for existing resources than any one member on its own. In addition, the effects of beneficial mutations of any one member are spread over the entire population. The formation of hypercycles has been suggested as an important step in the transition from inanimate to living chemistry, and a large number of hypercycles are expected to be embedded within the complex networks of living systems. But only one naturally occurring hypercycle has been well documented, while two autocatalytic chemical systems may contain vestiges of hypercyclic organization. Here we report a chemical system that constitutes a clear example of a minimal hypercyclic network, in which two otherwise competitive self replicating peptides symbiotically catalyse each others' production. PMID- 9403685 TI - Genomic sequence of a Lyme disease spirochaete, Borrelia burgdorferi. AB - The genome of the bacterium Borrelia burgdorferi B31, the aetiologic agent of Lyme disease, contains a linear chromosome of 910,725 base pairs and at least 17 linear and circular plasmids with a combined size of more than 533,000 base pairs. The chromosome contains 853 genes encoding a basic set of proteins for DNA replication, transcription, translation, solute transport and energy metabolism, but, like Mycoplasma genitalium, it contains no genes for cellular biosynthetic reactions. Because B. burgdorferi and M. genitalium are distantly related eubacteria, we suggest that their limited metabolic capacities reflect convergent evolution by gene loss from more metabolically competent progenitors. Of 430 genes on 11 plasmids, most have no known biological function; 39% of plasmid genes are paralogues that form 47 gene families. The biological significance of the multiple plasmid-encoded genes is not clear, although they may be involved in antigenic variation or immune evasion. PMID- 9403687 TI - Absence of contour linking in peripheral vision. AB - Human foveal vision is subserved initially by groups of spatial, temporal and orientational 'filters', the outputs of which are combined to define perceptual objects. Although a great deal is known about the filtering properties of individual cortical cells, relatively little is known about the nature of this 'linking' process. One recent approach has shown that the process can be thought of in terms of an association field whose strength is determined conjointly by the orientation and distance of the object. Here we describe a fundamental difference in this feature-linking process in central and peripheral parts of the visual field, which provides insight into the ways that foveal and peripheral visual perception differ. In the fovea, performance can be explained only by intercellular linking operations whereas in the periphery intracellular filtering will suffice. This difference represents a substantial economy in cortical neuronal processing of peripheral visual information and may allow a recent theory of intercellular binding to be tested. PMID- 9403688 TI - Fear conditioning induces associative long-term potentiation in the amygdala. AB - Long-term potentiation (LTP) is an experience-dependent form of neural plasticity believed to involve mechanisms that underlie memory formation. LTP has been studied most extensively in the hippocampus, but the relation between hippocampal LTP and memory has been difficult to establish. Here we explore the relation between LTP and memory in fear conditioning, an amygdala-dependent form of learning in which an innocuous conditioned stimulus (CS) elicits fear responses after being associatively paired with an aversive unconditioned stimulus (US). We have previously shown that LTP induction in pathways that transmit auditory CS information to the lateral nucleus of the amygdala (LA) increases auditory-evoked field potentials in this nucleus. Now we show that fear conditioning alters auditory CS-evoked responses in LA in the same way as LTP induction. The changes parallel the acquisition of CS-elicited fear behaviour, are enduring, and do not occur if the CS and US remain unpaired. LTP-like associative processes thus occur during fear conditioning, and these may underlie the long-term associative plasticity that constitutes memory of the conditioning experience. PMID- 9403689 TI - Fear conditioning induces a lasting potentiation of synaptic currents in vitro. AB - The amygdala plays a critical role in the mediation of emotional responses, particularly fear, in both humans and animals. Fear conditioning, a conditioned learning paradigm, has served as a model for emotional learning in animals, and the neuroanatomical circuitry underlying the auditory fear-conditioning paradigm is well characterized. Synaptic transmission in the medial geniculate nucleus (MGN) to lateral nucleus of the amygdala (LA) pathway, a key segment of the auditory fear conditioning circuit, is mediated largely through N-methyl-D aspartate (NMDA) and non-NMDA (such as alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA)) glutamate receptors; the potential for neural plasticity in this pathway is suggested by its capacity to support long-term potentiation (LTP). Here we report a long-lasting increase in the synaptic efficacy of the MGN-LA pathway attributable to fear-conditioning itself, rather than an electrically induced model of learning. Fear-conditioned animals show a presynaptic facilitation of AMPA-receptor-mediated transmission, directly measured in vitro with whole-cell recordings in lateral amygdala neurons. These findings represent one of the first in vitro measures of synaptic plasticity resulting from emotional learning by whole animals. PMID- 9403690 TI - How opioids inhibit GABA-mediated neurotransmission. AB - The midbrain region periaqueductal grey (PAG) is rich in opioid receptors and endogenous opioids and is a major target of analgesic action in the central nervous system. It has been proposed that the analgesic effect of opioids on the PAG works by suppressing the inhibitory influence of the neurotransmitter GABA (gamma-aminobutyric acid) on neurons that form part of a descending antinociceptive pathway. Opioids inhibit GABA-mediated (GABAergic) synaptic transmission in the PAG and other brain regions by reducing the probability of presynaptic neurotransmitter release, but the mechanisms involved remain uncertain. Here we report that opioid inhibition of GABAergic synaptic currents in the PAG is controlled by a presynaptic voltage-dependent potassium conductance. Opioid receptors of the mu type in GABAergic presynaptic terminals are specifically coupled to this potassium conductance by a pathway involving phospholipase A2, arachidonic acid and 12-lipoxygenase. Furthermore, opioid inhibition of GABAergic synaptic transmission is potentiated by inhibitors of the enzymes cyclooxygenase and 5-lipoxygenase, presumably because more arachidonic acid is available for conversion to 12-lipoxygenase products. These mechanisms account for the analgesic action of cyclooxygenase inhibitors in the PAG and their synergism with opioids. PMID- 9403691 TI - A Schwann cell mitogen accompanying regeneration of motor neurons. AB - Motor neurons are the only adult mammalian neurons of the central nervous system to regenerate following injury. This ability is dependent on the environment of the peripheral nerve and an intrinsic capacity of motor neurons for regrowth. We report here the identification, using a technique known as messenger RNA differential display, of an extracellular signalling molecule, previously described as the pancreatic secreted protein Reg-2, that is expressed solely in regenerating and developing rat motor and sensory neurons. Axon-stimulated Schwann cell proliferation is necessary for successful regeneration, and we show that Reg-2 is a potent Schwann cell mitogen in vitro. In vivo, Reg-2 protein is transported along regrowing axons and inhibition of Reg-2 signalling significantly retards the regeneration of Reg-2-containing axons. During development, Reg-2 production by motor and sensory neurons is regulated by contact with peripheral targets. Strong candidates for peripheral factors regulating Reg-2 production are cytokines of the LIF/CNTF family, because Reg-2 is not expressed in developing motor or sensory neurons of mice carrying a targeted disruption of the LIF receptor gene, a common component of the receptor complexes for all of the LIF/CNTF family. PMID- 9403692 TI - Role of cytosolic phospholipase A2 in allergic response and parturition. AB - Phospholipase A2 (PLA2) comprises a superfamily of enzymes that hydrolyse the ester bond of phospholipids at the sn-2 position. Among the members of this superfamily, cytosolic PLA2 has attracted attention because it preferentially hydrolyses arachidonoyl phospholipids and is activated by submicromolar concentrations of Ca2+ ions and by phosphorylation by mitogen-activated protein kinases (MAP kinases). Here we investigate the function of cytosolic PLA2 in vivo by using homologous recombination to generate mice deficient in this enzyme. These mice showed a marked decrease in their production of eicosanoids and platelet-activating factor in peritoneal macrophages. Their ovalbumin-induced anaphylactic responses were significantly reduced, as was their bronchial reactivity to methacholine. Female mutant mice failed to deliver offspring, but these could be rescued by administration of a progesterone-receptor antagonist to the mother at term. Considered together with previous findings, our results indicate that cytosolic PLA2 plays a non-redundant role in allergic responses and reproductive physiology. PMID- 9403693 TI - Reduced fertility and postischaemic brain injury in mice deficient in cytosolic phospholipase A2. AB - Phospholipase A2 (PLA2) enzymes are critical regulators of prostaglandin and leukotriene synthesis and can directly modify the composition of cellular membranes. PLA2 enzymes release fatty acids and lysophospholipids, including the precursor of platelet-activating factor, PAF, from phospholipids. Free fatty acids, eicosanoids, lysophospholipids and PAF are potent regulators of inflammation, reproduction and neurotoxicity. The physiological roles of the various forms of PLA2 are not well defined. The cytosolic form, cPLA2, preferentially releases arachidonic acid from phospholipids and is regulated by changes in intracellular calcium concentration. We have now created 'knockout' (cPLA2-/-) mice that lack this enzyme, in order to evaluate its physiological importance. We find that cPLA2-/- mice develop normally, but that the females produce only small litters in which the pups are usually dead. Stimulated peritoneal macrophages from cPLA2-/- animals did not produce prostaglandin E2 or leukotriene B4 or C4. After transient middle cerebral artery occlusion, cPLA2-/- mice had smaller infarcts and developed less brain oedema and fewer neurological deficits. Thus cPLA2 is important for macrophage production of inflammatory mediators, fertility, and in the pathophysiology of neuronal death after transient focal cerebral ischaemia. PMID- 9403694 TI - Miranda directs Prospero to a daughter cell during Drosophila asymmetric divisions. AB - Asymmetric cell division is a general process used in many developmental contexts to create two differently fated cells from a single progenitor cell. Intrinsic mechanisms like the asymmetric transmission of cell-fate determinants during cell division, and extrinsic cell-interaction mechanisms, can mediate asymmetric divisions. During embryonic development of the Drosophila central nervous system, neural stem cells called neuroblasts divide asymmetrically to produce another multipotent neuroblast and a ganglion mother cell (GMC) of more restricted developmental potential. Intrinsic mechanisms promote asymmetric division of neuroblasts: for example, the transcription factor Prospero localizes to the basal cell cortex of mitotic neuroblasts and then segregates exclusively into the GMC, which buds off from the basal side of the neuroblast. In the GMC, Prospero translocates to the nucleus, where it establishes differential gene expression between sibling cells. Here we report the identification of a gene, miranda, which encodes a new protein that co-localizes with Prospero in mitotic neuroblasts, tethers Prospero to the basal cortex of mitotic neuroblasts, directing Prospero into the GMC, and releases Prospero from the cell cortex within GMCs. miranda thus creates intrinsic differences between sibling cells by mediating the asymmetric segregation of a transcription factor into only one daughter cell during neural stem-cell division. PMID- 9403695 TI - Antiproliferative action of interferon-alpha requires components of T-cell receptor signalling. AB - Signal transduction through both cytokine and lymphocyte antigen receptors shares some common pathways by which they initiate cellular responses, such as activation of mitogen-activated protein kinase(s). However, other signalling components appear to be uniquely coupled to each receptor. For example, the interferon receptors transduce regulatory signals through the JAK/STAT pathway, resulting in an inhibition of growth and of antiviral effects, whereas this pathway apparently plays no role in T-cell-receptor (TCR)-dependent gene expression. Conversely, signal transduction through the TCR requires the tyrosine kinases Lck and ZAP-70 and the tyrosine phosphatase CD45. Here we show that, unexpectedly, transmission of growth-inhibitory signals by interferon-alpha (IFN alpha) in T cells requires the expression and association of CD45, Lck and ZAP-70 with the IFN-alpha-receptor signalling complex. PMID- 9403696 TI - Cdc42 and Rac1 induce integrin-mediated cell motility and invasiveness through PI(3)K. AB - Transformation of mammary epithelial cells into invasive carcinoma results in alterations in their integrin-mediated responses to the extracellular matrix, including a loss of normal epithelial polarization and differentiation, and a switch to a more motile, invasive phenotype. Changes in the actin cytoskeleton associated with this switch suggest that the small GTPases Cdc42 and Rac, which regulate actin organization, might modulate motility and invasion. However, the role of Cdc42 and Rac1 in epithelial cells, especially with respect to integrin mediated events, has not been well characterized. Here we show that activation of Cdc42 and Rac1 disrupts the normal polarization of mammary epithelial cells in a collagenous matrix, and promotes motility and invasion. This motility does not require the activation of PAK, JNK, p70 S6 kinase, or Rho, but instead requires phosphatidylinositol-3-OH kinase (PI(3)K). Further, direct PI(3)K activation is sufficient to disrupt epithelial polarization and induce cell motility and invasion. PI(3)K inhibition also disrupts actin structures, suggesting that activation of PI(3)K by Cdc42 and Rac1 alters actin organization, leading to increased motility and invasiveness. PMID- 9403697 TI - An extended microtubule-binding structure within the dynein motor domain. AB - Flagellar dynein was discovered over 30 years ago as the first motor protein capable of generating force along microtubules. A cytoplasmic form of dynein has also been identified which is involved in mitosis and a wide range of other intracellular movements. Rapid progress has been made on understanding the mechanism of force production by kinesins and myosins. In contrast, progress in understanding the dyneins has been limited by their great size (relative molecular mass 1,000K-2,000K) and subunit complexity. We now report evidence that the entire carboxy-terminal two-thirds of the 532K force-producing heavy chain subunit is required for ATP-binding activity. We further identify a microtubule binding domain, which, surprisingly, lies well downstream of the entire ATPase region and is predicted to form a hairpin-like stalk. Direct ultrastructural analysis of a recombinant fragment confirms this model, and suggests that the mechanism for dynein force production differs substantially from that of other motor proteins. PMID- 9403698 TI - Structure of the proteasome activator REGalpha (PA28alpha). AB - The specificity of the 20S proteasome, which degrades many intracellular proteins, is regulated by protein complexes that bind to one or both ends of the cylindrical proteasome structure. One of these regulatory complexes, the 11S regulator (known as REG or PA28), stimulates proteasome peptidase activity and enhances the production of antigenic peptides for presentation by class I molecules of the major histocompatibility complex (MHC). The three REG subunits that have been identified, REGalpha, REGbeta and REGgamma (also known as the Ki antigen), share extensive sequence similarity, apart from a highly variable internal segment of 17-34 residues which may confer subunit-specific properties. REGalpha and REGbeta preferentially form a heteromeric complex, although purified REGalpha forms a heptamer in solution and has biochemical properties similar to the heteromeric REGalpha/REGbeta complex. We have now determined the crystal structure of human recombinant REGalpha at 2.8 A resolution. The heptameric barrel-shaped assembly contains a central channel that has an opening of 20 A diameter at one end and another of 30 A diameter at the presumed proteasome binding surface. The binding of REG probably causes conformational changes that open a pore in the proteasome alpha-subunits through which substrates and products can pass. PMID- 9403699 TI - The hair follicle: dying for attention. PMID- 9403700 TI - Fibrous dysplasia of bone: the bone lesion unmasked. PMID- 9403701 TI - Expression of a virus-derived cytokine, KSHV vIL-6, in HIV-seronegative Castleman's disease. AB - Castleman's disease is a rare B cell lymphoproliferative disorder related to excess interleukin-6 (IL-6)-like activity. Kaposi's sarcoma-associated herpesvirus (KSHV or HHV8), which encodes a functional cytokine (vIL-6), has been found in some patients with Castleman's disease. Lymph nodes from 14 HIV seronegative Castleman's disease patients were compared to hyperplastic lymph nodes from 25 HIV-seronegative patients as well as Kaposi's sarcoma lesions from 48 patients for KSHV infection and vIL-6, human IL-6, and Epstein-Barr virus EBER expression. While all Kaposi's sarcoma tissues examined were polymerase chain reaction-positive and all control lymph nodes were polymerase chain reaction negative for KSHV, none had detectable vIL-6 expression. Six of 14 (43%) Castleman's tissues were positive for KSHV by polymerase chain reaction and all 6 had evidence of vIL-6 expression by immunohistochemistry. vIL-6-positive Castleman's disease patients generally had the multicentric plasma cell variant form of the disease and had a rapidly fatal clinical course frequently associated with autoimmune hemolytic anemia and gammopathy. In contrast, 7 (88%) of the 8 vIL-6-negative Castleman's disease patients had localized disease and have remained disease-free after therapy. KSHV vIL-6 expression appears to be limited to hematopoietic cells and is not present in Kaposi's sarcoma spindle cells. These data suggest that Castleman's disease is a syndrome of multiple etiologies involving aberrant IL-6 activity from either endogenous or viral sources. PMID- 9403702 TI - Neutralizing antibodies against epidermal growth factor and ErbB-2/neu receptor tyrosine kinases down-regulate vascular endothelial growth factor production by tumor cells in vitro and in vivo: angiogenic implications for signal transduction therapy of solid tumors. AB - The overexpression in tumor cells of (proto)-oncogenic receptor tyrosine kinases such as epidermal growth factor receptor (EGFR) or ErbB2/neu (also known as HER 2) is generally thought to contribute to the development of solid tumors primarily through their effects on promoting uncontrolled cell proliferation. However, agents that antagonize the function of the protein products encoded by these (proto)-oncogenes are known to behave in vivo in a cytotoxic-like manner. This implies that such oncogenes may regulate critical cell survival functions, including angiogenesis. The latter could occur as a consequence of regulation of relevant growth factors by such oncogenes. We therefore sought to determine whether EGFR or ErbB2/neu may contribute to tumor angiogenesis by examining their effects on the expression of vascular endothelial cell growth factor (VEGF)/vascular permeability factor (VPF), one of the most important of all known inducers of tumor angiogenesis. We found that in vitro treatment of EGFR-positive A431 human epidermoid carcinoma cells, which are known to be heavily dependent on VEGF/VPF in vivo as an angiogenesis growth factor, with the C225 anti-EGFR neutralizing antibody caused a dose-dependent inhibition of VEGF protein expression. Prominent suppression of VEGF/VPF expression in vivo, as well as a significant reduction in tumor blood vessel counts, were also observed in established A431 tumors shortly after injection of the antibody as few as four times into nude mice. Transformation of NIH 3T3 fibroblasts with mutant ErbB2/neu, another EGFR-like oncogenic tyrosine kinase, resulted in a significant induction of VEGF/VPF, and the magnitude of this effect was further elevated by hypoxia. Moreover, treatment of ErbB2/neu-positive SKBR-3 human breast cancer cells in vitro with a specific neutralizing anti-ErbB2/neu monoclonal antibody (4D5) resulted in a dose-dependent reduction of VEGF/VPF protein expression. Taken together, the results suggest that oncogenic properties of EGFR and ErbB2/neu may, at least in part, be mediated by stimulation of tumor angiogenesis by up-regulating potent angiogenesis growth factors such as VEGF/VPF. These genetic changes may cooperate with epigenetic/environmental effects such as hypoxia to maximally stimulate VEGF/VPF expression. Therapeutic disruption of EGFR or ErbB2/neu protein function in vivo may therefore result in partial suppression of angiogenesis, a feature that could enhance the therapeutic index of such agents in vivo and endow them with anti-tumor effects, the magnitude of which may be out of proportion with their observed cytostatic effects in monolayer tissue culture. PMID- 9403703 TI - Molecular aberrations of the G1-S checkpoint in myxoid and round cell liposarcoma. AB - Myxoid and round cell liposarcoma represents a morphological spectrum in which tumor progression from low-grade myxoid to high-grade round cell areas is frequently observed. A distinctive t(12;16)(q13;p11) reciprocal translocation rearranges the CHOP gene localized to 12q13 in most cases. Data concerning the occurrence of cell cycle aberrations in this subset of mesenchymal malignancies are very limited. Therefore, we analyzed a histologically homogeneous series of 21 cases of myxoid and round cell liposarcoma. The p53 pathway was studied by investigating the TP53 gene and protein, mdm2 protein, and p21Waf1 protein. The Rb-cyclin D pathway was analyzed by studying the pRb protein, the p16MTS1 gene, cyclin D1, cyclin D3, p27Kip1, cdk4, and cdk6 proteins. In contrast with the rare involvement of the TP53 gene in well differentiated liposarcoma, aberrations of the TP53 gene were observed in approximately 30% of cases of myxoid and round cell liposarcoma. Notably, mdm2 overexpression was seen in 56% of cases and correlated with histological grade, therefore indicating a possible role in tumor progression. Abnormalities involving the Rb-cyclin D pathway were observed in more than 90% of cases. pRb loss was present in one-third of cases and, at variance with that observed in other subsets of sarcoma, overexpression of cyclin Ds represented a rare event. Interestingly, upregulation of either cdk4 or cdk6 was demonstrated in 85% of cases. PMID- 9403704 TI - The FHIT gene product is highly expressed in the cytoplasm of renal tubular epithelium and is down-regulated in kidney cancers. AB - Loss of heterozygosity and homozygous deletion of the 3p14.2 region in human cancers implies the existence of a tumor suppressor gene. One such candidate is the fragile histidine triad (FHIT) gene. To investigate the role of FHIT gene product in tumorigenesis, we generated specific polyclonal antibodies to the human protein and studied its expression in normal and tumor tissues. Immunoblot analysis revealed highly variable expression of pFhit in normal adult human tissues. The highest steady-state level of pFhit was found in kidney and brain, whereas breast, intestine, and skeletal muscle expressed only trace amounts. Within the kidney, the pattern of pFhit immunoreactivity was confined to the tubular epithelium and absent in the glomeruli. Immunofluorescence analysis and biochemical fractionation have sublocalized pFhit to the cytosolic compartment. Compared with normal kidney, pFhit was found to be down-regulated in a subset of primary renal cell carcinoma. Two of 12 renal cell carcinoma cell lines that are known not to contain VHL mutations showed complete loss of pFhit expression. This is supported by the appearance of aberrant reverse transcription-polymerase chain reaction products and loss of the normal-size fragment. Our results are consistent with a potential role of pFhit loss or dysfunction in human renal cell carcinoma independent of VHL involvement. PMID- 9403705 TI - MAGE-3 immunoreactivity in formalin-fixed, paraffin-embedded primary and metastatic melanoma: frequency and distribution. AB - Monoclonal antibody 57B specifically detects MAGE-3 gene protein expression. MAGE derived peptides are recognized by CD8+ T cells and applied in immunotherapy. We examined formalin-fixed, paraffin-embedded tissue of 61 melanoma (primary, n = 40; metastatic, n = 21) and 46 control cases (junctional, dermal, compound, Spitz, Reed, and balloon-cell nevi) by immunohistochemistry using the alkaline phosphatase anti-alkaline phosphatase method after antigen retrieval. Immunoreactivity was rated positive at 20 positive cells per tumor or more. Staining pattern was homogeneous, scattered, or focal. All control samples and internal controls were immunonegative. Staining with monoclonal antibody 57B showed a specificity of 100% with a sensitivity of 44%. Immunopositivity (overall, 44% of melanomas) increased along with tumor, node, and metastasis stage; pT1 showed 13%, pT2 22%, pT3a 29%, pT3b 45%, pT4 100%, pTxN1 60%, and pTxNxM1a 63% of samples positive. The staining pattern was homogeneous on pT1 to pT3a tumors, homogeneous or focal in pT3b and pT4a, and homogeneous, focal, or scattered in pTxN1 and pTxNxM1a. The frequency of immunopositivity relates well to data on mRNA expression using reverse transcriptase polymerase chain reaction in a subgroup analyzed by both methods. Monoclonal antibody 57B can be used to allow profiling of melanomas using routine archival tissue, when considering immunotherapeutic approaches involving MAGE-3-derived epitopes. PMID- 9403706 TI - Matrix vesicles in osteomalacic hypophosphatasia bone contain apatite-like mineral crystals. AB - Hypophosphatasia, a heritable disease characterized by deficient activity of the tissue nonspecific isoenzyme of alkaline phosphatase (TNSALP), results in rickets and osteomalacia. Although identification of TNSALP gene defects in hypophosphatasia establishes a role of ALP in skeletal mineralization, the precise function remains unclear. The initial site of mineralization (primary mineralization) normally occurs within the lumen of TNSALP-rich matrix vesicles (MVs) of growth cartilage, bone, and dentin. We investigated whether defective calcification in hypophosphatasia is due to a paucity and/or a functional failure of MVs secondary to TNSALP deficiency. Nondecalcified autopsy bone and growth plate cartilage from five patients with perinatal (lethal) hypophosphatasia were studied by nondecalcified light and electron microscopy to assess MV numbers, size, shape, and ultrastructure and whether hypophosphatasia MVs contain apatite like mineral, as would be the case if these MVs retained their ability to concentrate calcium and phosphate internally despite a paucity of TNSALP in their investing membranes. We found that hypophosphatasia MVs are present in approximately normal numbers and distribution and that they are capable of initiating internal mineralization. There is retarded extravesicular crystal propagation. Thus, in hypophosphatasia the failure of bones to calcify appears to involve a block of the vectorial spread of mineral from initial nuclei within MVs, outwards, into the matrix. We conclude that hypophosphatasia MVs can concentrate calcium and phosphate internally despite a deficiency of TNSALP activity. PMID- 9403707 TI - Acute pulmonary toxicity of urban particulate matter and ozone. AB - We have investigated the acute lung toxicity of urban particulate matter in interaction with ozone. Rats were exposed for 4 hours to clean air, ozone (0.8 ppm), the urban dust EHC-93 (5 mg/m3 or 50 mg/m3), or ozone in combination with urban dust. The animals were returned to clean air for 32 hours and then injected (intraperitoneally) with [3H]thymidine to label proliferating cells and killed after 90 minutes. The lungs were fixed by inflation, embedded in glycol methacrylate, and processed for light microscopy autoradiography. Cell labeling was low in bronchioles (0.14 +/- 0.04%) and parenchyma (0.13 +/- 0.02%) of air control animals. Inhalation of EHC-93 alone did not induce cell labeling. Ozone alone increased (P < 0.05) cell labeling (bronchioles, 0.42 +/- 0.16%; parenchyma, 0.57 +/- 0.21%), in line with an acute reparative cell proliferation. The effects of ozone were clearly potentiated by co-exposure with either the low (3.31 +/- 0.31%; 0.99 +/- 0.18%) or the high (4.45 +/- 0.51%; 1.47 +/- 0.18%) concentrations of urban dust (ozone X EHC-93, P < 0.05). Cellular changes were most notable in the epithelia of terminal bronchioles and alveolar ducts and did not distribute to the distal parenchyma. Enhanced DNA synthesis indicates that particulate matter from ambient air can exacerbate epithelial lesions in the lungs. This may extend beyond air pollutant interactions, such as to effects of inhaled particles in the lungs of compromised individuals. PMID- 9403708 TI - Central nervous system lipoproteins in Alzheimer's disease. AB - Alzheimer's disease (AD) is the most common form of dementia in the United States and has been associated with APOE genotype. Apolipoprotein (apo) E along with apoAI serve as the major apolipoproteins in the central nervous system; however, we are unaware of any study addressing lipoprotein metabolism in AD. We tested the hypothesis that lipoprotein metabolism is altered in patients with AD by isolating and characterizing ventricular fluid (VF) lipoproteins obtained during a rapid autopsy protocol from patients with AD and age-matched nondemented control patients. Our results demonstrated abnormalities in the protein and lipid constituents of VF lipoproteins from AD patients. Apolipoprotein concentration was reduced by half in AD patients relative to controls; however, there was no selective reduction in apoE or apoAI. In addition, cholesteryl ester fatty acids, but not phospholipid fatty acids, from AD patients demonstrated a significant reduction in some polyunsaturated fatty acids (18:2 and 22:6) and an enrichment in 18:0. None of these changes were directly related to APOE genotype. Our data indicate that VF lipoprotein composition is altered, at least terminally, in AD patients, and that these changes are not associated with APOE. These findings suggest that altered VF lipoprotein metabolism may be a component of AD pathogenesis. PMID- 9403709 TI - CNA.42, a new monoclonal antibody directed against a fixative-resistant antigen of follicular dendritic reticulum cells. AB - A new monoclonal antibody (MAb), CNA.42, was generated using the CEM T-cell line. It recognizes a 120-kd formalin-resistant glycosylated antigen that is mainly expressed by follicular dendritic reticulum cells (FDRCs). This antigen is also expressed by a few mononuclear cells in the paracortical area of reactive lymph nodes and by some cortical thymocytes. Two hundred and eighty-nine cases of hematopoietic tumors of various types were tested with this antibody. They showed either intact FDRC networks or FDRC networks dispersed among malignant cells. In follicular lymphomas, the follicular pattern was highlighted by CNA.42 MAb. Expanded FDRC networks were found in angioimmunoblastic T-cell lymphomas. Neoplastic cells were positive in 43.6% (24/55) of T-cell and 4.6% (6/129) of B cell lymphomas. The highest percentage of cases with positive neoplastic cells was found in anaplastic large-cell lymphomas (62.5%; 15/24). In Hodgkin's disease, FDRC networks, sometimes encasing Hodgkin and Reed-Sternberg (HRS) cells, were found. HRS cells were also stained by this antibody in 23 (21.9%) of the 105 cases examined. A variety of normal nonlymphoid tissues and nonhematopoietic tumors, such as some neurogenic tumors, carcinoma, and occasional sarcomas, were found to be positive. Analysis of the reactivity of CNA.42 antibody with FDRCs of lymphoid tissue from different animal species showed similar reactivity to that observed in humans, suggesting widespread evolutionary conservation of the antigen recognized by this antibody. In daily diagnostic practice, CNA.42 MAb seems to be a suitable FDRC marker and possibly has an auxiliary role in recognizing T-cell lymphomas. PMID- 9403710 TI - Fibrous dysplasia of bone in the McCune-Albright syndrome: abnormalities in bone formation. AB - In addition to cafe-au-lait pigmentation patterns and hyperendocrinopathies, fibrous dysplasia of bone is a major finding in the McCune-Albright syndrome. Activating missense mutations of the Gs alpha gene leading to overactivity of adenylyl cyclase have been identified in patients with McCune-Albright syndrome, but the mechanism leading to the specific development of fibrous dysplasia in bone has not been elucidated. By means of specific peptide antisera and reverse transcriptase polymerase chain reaction in situ hybridization, we show that expression of Gs alpha and its mRNA is critically up-regulated during maturation of precursor osteogenic cells to normal osteoblast cells and that this pattern of expression is retained in fibrous dysplasia. A functional characterization of fibrous dysplastic tissues revealed that the fibrotic areas consist, in fact, of an excess of cells with phenotypic features of pre-osteogenic cells, whereas the lesional bone formed de novo within fibrotic areas represents the biosynthetic output of mature but abnormal osteoblasts. These cells are noted for peculiar changes in cell shape and interaction with matrix, which were mimicked in vitro by the effects of excess exogenous cAMP on human osteogenic cells. Osteoblasts involved with the de novo deposition of lesional bone in fibrous dysplasia produce a bone matrix enriched in certain anti-adhesion molecules (versican and osteonectin), and poor in the pro-adhesive molecules osteopontin and bone sialoprotein, which is in contrast to the high levels of these two proteins found in normal de novo bone. Our data indicate the need to reinterpret fibrous dysplasia of bone as a disease of cells in the osteogenic lineage, related to the effects of excess cAMP on bone cell function. They further suggest that a critical, physiological, maturation-related regulation of Gs alpha levels makes cells in the osteogenic lineage a natural target for the effects of mutations in the Gs alpha gene and may provide a clue as to why bone itself is affected in this somatic, mutation-dependent disease. PMID- 9403711 TI - Analysis of apoptosis during hair follicle regression (catagen) AB - Keratinocyte apoptosis is a central element in the regulation of hair follicle regression (catagen), yet the exact location and the control of follicular keratinocyte apoptosis remain obscure. To generate an "apoptomap" of the hair follicle, we have studied selected apoptosis-associated parameters in the C57BL/6 mouse model for hair research during normal and pharmacologically manipulated, pathological catagen development. As assessed by terminal deoxynucleotide transferase dUTP fluorescein nick end-labeling (TUNEL) stain, apoptotic cells not only appeared in the regressing proximal follicle epithelium but, surprisingly, were also seen in the central inner root sheath, in the bulge/isthmus region, and in the secondary germ, but never in the dermal papilla. These apoptosis hot spots during catagen development correlated largely with a down-regulation of the Bcl 2/Bax ratio but only poorly with the expression patterns of interleukin-1beta converting enzyme, p55TNFR, and Fas/Apo-1 immunoreactivity. Instead, a higher correlation was found with p75NTR expression. During cyclophosphamide-induced follicle dystrophy and alopecia, massive keratinocyte apoptosis occurred in the entire proximal hair bulb, except in the dermal papilla, despite a strong up regulation of Bax and p75NTR immunoreactivity. Selected receptors of the tumor necrosis factor/nerve growth factor family and members of the Bcl-2 family may also play a key role in the control of follicular keratinocyte apoptosis in situ. PMID- 9403713 TI - Diverse cell death pathways result from a single missense mutation in weaver mouse. AB - Neuronal death affects selectively granule cell precursors of the cerebellum and the dopaminergic neurons of midbrain in the weaver mutant mouse. The weaver phenotype is associated with a missense mutation in the gene coding for the GIRK2 potassium channel, which results in chronic depolarization. Using DNA gel electrophoresis, electron microscopy (EM), the in situ end-labeling (ISEL) technique at the light and EM level, and immunohistochemistry for apoptosis related proteins c-Jun and proliferating cell nuclear antigen (PCNA), we have investigated the mechanisms of cell death in cerebellum and substantia nigra. Between postnatal day P1 and P21, in the external germinal layer of the cerebellum, most degenerating granule cell precursors were found to aggregate to form clusters. Degenerating cells exhibited strong nuclear staining for ISEL, c Jun, and PCNA and had a typical apoptotic morphology by EM. Increased c-Jun and ISEL staining were also occasionally seen in Purkinje cells. Between P14 and P21, when dopaminergic neurons start to degenerate, staining for ISEL, c-Jun, and PCNA in weaver substantia nigra was the same as in controls. By EM, however, we found only in weaver mice numerous dopaminergic cells that showed extensive vacuolar and autophagic changes of cytoplasm, preservation of membrane and organelle integrity, and absence of chromatin condensation or DNA fragmentation by EM-ISEL. The combination of vacuolar and autophagic changes identifies a novel type of non necrotic, nonapoptotic cell death. After biochemical analysis of DNA, a clear-cut laddering, suggestive of oligonucleosomal fragmentation, was present in samples from weaver cerebellum. Cell death diversity appears to be influenced by specific features of target cells. These findings may be relevant for understanding the mechanisms of cell death in neurodegenerative diseases. PMID- 9403712 TI - Altered expression of keratinocyte growth factor and its receptor in psoriasis. AB - One of the biological characteristics of psoriasis is excessive flaking of the skin. This is directly related to the marked hyperplasia of epidermal keratinocytes and to incomplete epidermal differentiation. Keratinocyte growth factor (KGF), a potent mitogen for human keratinocytes, is expressed by stromal cells. Alterations in the KGF signaling pathway might account for the epidermal hyperplasia associated with psoriasis. To test this hypothesis, we investigated the expression of KGF and its receptor (KGFR) in psoriasis tissue. KGF and KGFR mRNA levels were found to be frequently elevated in psoriatic skin specimens as compared with normal skin. Increased KGF transcript expression was localized to the dermal layer of the involved skin specimen using in situ hybridization. In contrast, KGFR transcript and protein expression was localized to the basal layer of keratinocytes in normal skin and to the basal and suprabasal layers of the psoriatic epidermis, coincident with the expanded proliferative keratinocyte pool. To identify molecules that might regulate KGFR expression we investigated the effects of various pharmacological agents and cytokines on KGFR synthesis by keratinocytes. Phorbol ester, interleukin-6, interferon-gamma, and ultraviolet B (UVB) treatment all led to substantial down-regulation of KGFR expression. The down-regulation of KGFR synthesis by UVB suggests a possible mechanism for the antiproliferative action of this agent in the treatment of psoriasis. Taken together, these results suggest that increased KGFR-mediated signaling in keratinocytes in the lesional epidermis might account in part for the epidermal hyperplasia in psoriasis. PMID- 9403714 TI - Loss of tuberin in both subependymal giant cell astrocytomas and angiomyolipomas supports a two-hit model for the pathogenesis of tuberous sclerosis tumors. AB - Tuberous sclerosis complex (TSC) is an autosomal dominant disorder characterized by seizures, mental retardation, and tumors of skin, brain, heart, and kidney. In this study, we focused on two of the most frequent tumors in TSC patients, renal angiomyolipomas and subependymal giant cell astrocytomas (SEGAs). Two questions were addressed. First, is loss of tuberin, the product of the TSC2 gene, seen in both renal and central nervous system tumors from TSC patients? Second, when loss of tuberin occurs, does it affect each of the cell types seen in these tumors? We used a loss of heterozygosity approach to identify tumors from TSC2 patients. We found loss of tuberin immunostaining in the spindle and epithelioid cells but not in the giant cells of six TSC2 SEGAs. We also found loss of tuberin immunostaining in all three cell types (smooth muscle, fat, and vessels) of six TSC2 angiomyolipomas. Chromosome 16p13 loss of heterozygosity occurred in both spindle and epithelioid cells of a SEGA and in smooth muscle and fat but not the vessels of two angiomyolipomas. These results support a two-hit tumor suppressor model for the pathogenesis of SEGAs and angiomyolipomas. The vascular elements of angiomyolipomas and the giant cells of SEGAs may be reactive rather than neoplastic. PMID- 9403716 TI - Organ-site dependence for the production of urokinase-type plasminogen activator and metastasis by human renal cell carcinoma cells. AB - We examined the role of urokinase-type plasminogen activator (u-PA) in the metastasis of the human renal cell carcinoma (HRCC) implanted in athymic nude mice. Cells from a HRCC KG-2 line were implanted in orthotopic (kidney) and ectopic (subcutaneous) organs. The KG-2 cells implanted in the kidney produced local tumors and lung metastases, whereas those implanted subcutaneously produced only local tumors. The production of u-PA was determined by immunohistochemistry and an enzyme-linked immunosorbent assay (ELISA). High levels of u-PA were produced by the metastatic kidney tumors and lung metastases, whereas the subcutaneous tumors produced low levels. KG-2 cells co-cultured with mouse kidney or lung fibroblasts produced higher levels of u-PA than KG-2 cells co-cultured with mouse skin fibroblasts. Furthermore, KG-2 cells cultured with the conditioned medium from mouse kidney or lung fibroblasts produced higher levels of u-PA than KG-2 cells cultured with the conditioned medium from mouse skin fibroblasts. The results indicate that the expression of u-PA by KG-2 cells is one of the important factors that determine their metastatic potential and that the production of u-PA is influenced by the organ microenvironment, including soluble factors produced by surrounding fibroblasts. PMID- 9403715 TI - Universal absence of merlin, but not other ERM family members, in schwannomas. AB - NF2 (neurofibromatosis 2, encoding the merlin protein) gene mutations and chromosome 22q loss have been demonstrated in the majority of sporadic and NF2 associated schwannomas, but many schwannomas fail to demonstrate genetic evidence of biallelic NF2 gene inactivation. In addition, the role of the merlin-related ERM family members (ezrin, radixin, and moesin) remains unclear in these tumors. We therefore studied expression of NF2-encoded merlin as well as ezrin, radixin, and moesin in 22 vestibular and peripheral schwannomas that had been evaluated for NF2 mutations and chromosome 22q loss. Western blotting and immunohistochemistry with antibodies directed against the amino and carboxy termini of merlin demonstrated loss of merlin expression in all studied schwannomas, including 12 tumors lacking genetic evidence of biallelic NF2 gene inactivation. Western blotting with antibodies directed against ezrin, radixin, and moesin, however, showed expression of these proteins in all schwannomas. In addition, immunohistochemistry with an antibody to moesin revealed widespread expression in tumor and endothelial cells. These data indicate that the specific loss of merlin is universal to schwannomas and is not linked to loss of ezrin, radixin, or moesin expression. PMID- 9403717 TI - Sp1-mediated transactivation of LamC1 promoter and coordinated expression of laminin-gamma1 and Sp1 in human hepatocellular carcinomas. AB - The laminin-gamma1 chain is present in most basement membranes and is involved in various physiological and pathological processes, including carcinogenesis in the liver. We have investigated the role of the transcription factor Sp1 in the activation of the LamC1 gene, which encodes laminin-gamma1, both in hepatocytes and in human hepatocellular carcinomas. DNAse I hypersensitive sites were mapped in the murine LamC1 promoter using early hepatocyte primary cultures in which LamC1 becomes activated. Three hypersensitive sites were found in enhancer-like elements that contain GC-rich regions. Gel-shift analyses showed that specific complexes were resolved using GC-containing oligonucleotides and Faza 567 hepatoma cells, which constitutively express laminin-gamma1 at a high level. Increased GC-binding activity was observed using nuclear extracts from early hepatocyte cultures versus normal liver. Sp1 overexpression in normal hepatocytes transfected with an Sp1 expression vector induced a marked increased of laminin gamma1 mRNA content and co-transfection of promoter fragments in Drosophila melanogaster SL2 cells demonstrated that Sp1 transactivates LamC1. In human hepatocellular carcinomas, Sp1 and laminin-gamma1 mRNA were simultaneously expressed at high levels, and gel-shift experiments demonstrated a higher GC binding activity to Sp1 compared with control livers. In situ hybridization indicated that cells exhibiting a high content of laminin-gamma1 mRNA were also strongly positive for Sp1 mRNA, including both cancer cells at the invasion front and stromal cells. These results show that Sp1 is involved in the activation of LamC1 that occurs in human hepatocellular carcinomas. PMID- 9403718 TI - Simple epithelium keratins are required for maintenance of hepatocyte integrity. AB - Keratin 8 (K8)-deficient adult mice develop a severe disease of the gastrointestinal tract characterized mainly by colorectal hyperplasia and inflammation. Given that hepatocytes contain K8/K18 heteropolymers only, this animal model was used to assess the contribution of these simple epithelium keratins to hepatocyte structural and functional integrity. Homozygous mutant (HMZ), heterozygous, and wild-type (WT) mice were examined for hepatocyte structural and metabolic features and their survival to partial hepatectomy. Except for the presence of few necrotic foci, no other tissular or cellular alterations were observed in nonhepatectomized HMZ mouse livers; glycogen and lipid peroxidation levels were essentially normal, but a small reduction in bile flow was observed. In response to a single pentobarbital injection, HMZ mice had longer sleeping times than heterozygous and WT mice. After a two-thirds partial hepatectomy under pentobarbital anesthesia, all HMZ mice died within a few hours, whereas those anesthetized with ether survived for 1 to 2 days. One hour after hepatectomy after pentobarbital anesthesia, many hepatocytes contained erythrocytes and large vacuoles in the cytoplasm, which suggests damage at the plasma membrane level in response to a sudden increase in portal blood flow. In line with these findings, an uptake of trypan blue by HMZ but not WT mouse hepatocytes was observed during a 10 ml/minute perfusion via the portal vein with a dye-supplemented buffer. Subsequent cellular dispersion led to viable WT mouse hepatocytes but largely nonviable HMZ mouse hepatocytes. Better viability was obtained at lower perfusion rates. Partially hepatectomized heterozygous mice developed liver steatosis, a condition that was not associated with a change in K8 content but perhaps linked to the presence of the neo gene. Transgenic HMZ mouse rescue experiments with a full-length K8 gene confirmed that the phenotypic alterations observed in partially hepatectomized HMZ mice were caused by the disruption of the K8 gene. Taken together, these findings demonstrate that simple epithelium keratins are essential for the maintenance of hepatocyte structural and functional integrity. PMID- 9403719 TI - NEU overexpression in the furan rat model of cholangiocarcinogenesis compared with biliary ductal cell hyperplasia. AB - Immunohistochemical studies have suggested that the tyrosine kinase growth factor receptor p185neu is overexpressed in a high percentage of human cholangiocarcinomas. To establish the specificity and temporal relationship between the expression of this receptor in cholangiocarcinogenesis, we investigated c-neu expression in precancerous cholangiofibrotic tissue and subsequently derived primary and transplantable cholangiocarcinomas originated in the livers of furan-treated rats. Proliferated bile ductules formed in rat models of bile ductular hyperplasia and the cell types of normal adult rat liver were also analyzed for c-neu expression. c-neu expression was not detected in normal adult rat liver by either Western blotting, immunohistochemistry, or in situ hybridization. In comparison, all of the cholangiocarcinomas analyzed, which were characterized by intestinal-type mucin-producing neoplastic glands, exhibited a prominent band with a molecular weight 185 kd, corresponding to p185neu. Only the neoplastic glandular epithelia of the cholangiocarcinomas showed a strong immunoreactivity for p185neu, which was predominantly localized to their cell surface but also observed cytoplasmically. In situ hybridization further revealed the cytoplasm of the tumor glandular epithelial cells to be strongly positive for c-neu mRNA transcripts. Of particular interest was our finding that c-neu is expressed early in furan cholangiocarcinogenesis, being more pronounced in the metaplastic intestinal glands of cholangiofibrotic tissue than in hyperplastic biliary epithelial cells in either the same tissue or in hyperplastic bile ductule tissue. Our results demonstrate that c-neu overexpression is a prominent feature of intestinal-type cholangiocarcinomas as well as of metaplastic intestinal glands that precede their development and is detected at lower levels in hyperplastic biliary epithelia. The overexpression of c-neu in the metaplastic and malignant neoplastic glands also correlated with their increased proliferating cell nuclear antigen (PCNA) labeling indices relative to those of hyperplastic biliary ducts and ductules and also appeared to correlate with their intestinal glandular pattern of differentiation. PMID- 9403720 TI - Increase in proliferation and apoptosis of gastric epithelial cells early in the natural history of Helicobacter pylori infection. AB - Childhood acquisition of Helicobacter pylori is a critical risk factor for gastric cancer. Since tumorigenesis involves deregulation of proliferation and apoptosis, we examined gastric epithelial cell proliferation and apoptosis in H. pylori-infected children. Apoptosis and proliferation of gastric antral epithelial cells in biopsy specimens from patients with H. pylori-induced gastritis, secondary gastritis, and noninflamed controls were compared. p53 protein expression was examined immunohistochemically. Apoptotic cells were identified in the surface epithelium in each group. The apoptotic index was higher in specimens from patients with H. pylori gastritis (120 +/- 10) than secondary gastritis (50 +/- 10) and noninflamed controls (40 +/- 10, analysis of variance P < 0.005). Apoptosis decreased following H. pylori eradication and resolution of gastritis (P < 0.02). An expanded proliferative compartment was identified in H. pylori-induced gastritis (32.4 +/- 3.5; proliferative labeling index +/- SE) compared with secondary gastritis (18.9 +/- 2.8) and noninflamed controls (13.7 +/- 3.1, analysis of variance P < 0.01). The accelerated cell turnover was associated with p53 overexpression (analysis of variance P < 0.005). Accumulation of p53 was not associated with expression of the cyclin-dependent kinase inhibitor p21. The occurrence of altered cell turnover early in the natural history of chronic infection provides an explanation for the increased risk of gastric cancer development associated with childhood acquisition of infection. PMID- 9403721 TI - Chronic cyclosporin A nephrotoxicity, P-glycoprotein overexpression, and relationships with intrarenal angiotensin II deposits. AB - P-glycoprotein (P-gp) expels hydrophobic substances from the cell, including chemotherapeutic agents and immunosuppressants such as cyclosporin A (CsA) and FK506. Exposure of cultured renal tubular cells to CsA induces P-gp overexpression in cell membranes. Angiotensin II has recently been implicated as the principal factor responsible for progression of interstitial fibrosis induced by CsA. To investigate the in vivo relationships between histological lesions, P gp overexpression, and intrarenal angiotensin II deposits, we developed a model of chronic CsA toxicity in Sprague-Dawley rats treated with 25 mg/kg/day CsA for 28 and 56 days and fed either a standard maintenance diet or a low-salt diet. Immunohistochemical methods were used to study the expression of P-gp in renal tubular cells and the appearance of intrarenal angiotensin II deposits. Rats treated with CsA developed chronic nephrotoxicity lesions that were more evident in the group fed the low-salt diet. Treatment with CsA induced overexpression of P-gp in tubular cells of the kidney that increased with time. We found that immunohistochemical expression of P-gp was slightly more severe in rats fed a low salt diet. Intrarenal deposits of angiotensin II were more evident in rats treated with CsA; these deposits also increased with time. This finding was also more relevant in rats given the low-salt diet. The up-regulation of P-gp was inversely related to the incidence of hyaline arteriopathy (r = -0.65; P < 0.05), periglomerular (r = -0.58; P < 0.05) and peritubular fibrosis (r = -0.63; P < 0.05), and intrarenal angiotensin H deposits in animals with severe signs of nephrotoxicity (r = -0.65; P < 0.05). These results support the hypothesis that the role of P-gp as a detoxicant in renal cells may be related to mechanisms that control the cytoplasmic removal of both toxic metabolites from CsA and those originating from the catabolism of signal transduction proteins (methylcysteine esters), which are produced as a result of ras activation in presence of angiotensin II. PMID- 9403722 TI - Lymphocyte recruitment and the kinetics of adhesion receptor expression during the pulmonary immune response to particulate antigen. AB - The selectins and beta2 integrins participate in the recruitment of neutrophils in acute pulmonary inflammation. However, the cell adhesion receptors that mediate lymphocyte trafficking into the lung have not been defined. This study examined the relationship between cell adhesion molecules on the pulmonary vasculature and on lymphocytes recovered from the lung during a pulmonary immune response to intratracheal (I.T.) sheep red blood cells (SRBCs) in sensitized C57BL/6J mice. Silver-enhanced immunogold staining and reverse transcriptase polymerase chain reaction of lung tissues revealed sustained induction of VCAM-1, E-selectin, and P-selectin on the pulmonary vasculature for up to 7 days after I.T.-SRBC challenge. Neither the MECA 79 nor MECA 367 antigens were induced on the pulmonary vasculature during this period. In the peripheral blood, both CD4 and CD8 T-cell subsets showed an initial increase in P-selectin ligand expression after I.T.-SRBC challenge. The number of P-selectin ligand-positive T cells in the peripheral blood fell as T cells with both P-selectin and, to a lesser extent, E-selectin ligands accumulated in the bronchoalveolar lavage fluid. We conclude that I.T.-SRBC challenge in sensitized mice elicits prolonged synthesis of P-selectin, E-selectin, and VCAM-1 by the lung vasculature as well as selectin ligand synthesis by responding T cells. Furthermore, the entry of selectin-ligand positive T cells into the circulation and their accumulation in the bronchoalveolar lavage fluid indicates that these receptors may contribute to T cell recruitment. Finally, VCAM-1 on the vasculature may also participate; however, the vascular addressins, required for homing to peripheral and mucosal lymphoid organs, are not essential for T-cell entry into the lung following I.T. SRBC challenge. PMID- 9403723 TI - Biochemical and immunohistochemical characterization of human type XIX defines a novel class of basement membrane zone collagens. AB - Nineteen types, the product of 33 genes, comprise the collagen family of proteins. Types I, II, III, V, and XI constitute the fibrillar collagens, whereas types IV, VI to X, and XII to XIX represent the structurally diverse, nonfibrillar members. Type XIX collagen was discovered from the sequence of rhabdomyosarcoma cDNA clones. The type XIX chain consists of 1142 amino acids that contribute primarily to a unique five subdomain triple-helical region. To characterize the protein, to determine the tissue distribution, and to provide some insight into its function, we generated two type XIX-specific polyclonal antibodies. One was directed against a recombinant molecule containing amino terminal sequences, and the second was derived from a synthetic peptide corresponding to most of the short carboxy terminus. These antibodies were used in immunoblot assays of rhabdomyosarcoma cell/matrix homogenates to identify a 165-kd disulfide-bonded and bacterial collagenase-sensitive protein. Immunohistochemical analysis of type XIX collagen was performed for human skeletal muscle, spleen, prostate, kidney, liver, placenta, colon, and skin. In contrast to Northern blot hybridizations, which showed very low levels of the 12 kb transcript in few tissues, the protein was found in all tissues examined. The type XIX collagen distribution was restricted to vascular, neuronal, mesenchymal, and some epithelial basement membrane zones, which is similar to the profile recently established (Ref. 8) and further extended here for type XV collagen. Nevertheless, localization of type XIX exhibited significant differences from type XV collagen that were particularly evident in the kidney, liver, and spleen. This report, in conjunction with the type XV results and other studies of type XVIII collagen, indicates the existence of a new collagen subgroup founded on their widespread presence in basement membrane zones regardless of chain homology. In addition to their role in basement membrane-stromal interactions, the pronounced vascular association suggests involvement of these related collagen types with angiogenic and pathological processes. PMID- 9403724 TI - Cellular retinol-binding protein-1 is transiently expressed in granulation tissue fibroblasts and differentially expressed in fibroblasts cultured from different organs. AB - We have reported that cellular retinol-binding protein-1 (CRBP-1) is transiently expressed by arterial smooth muscle cells during experimental intimal repair (P. Neuville, A. Geinoz, G. Benzonana, M. Redard, F. Gabbiani, P. Ropraz, G. Gabbiani: Am J Pathol 1997, 150:509-521). We have examined here the expression of CRBP-1 during wound healing after a full-thickness rat skin wound. CRBP-1 was transiently expressed by a significant proportion of fibroblastic cells including myofibroblasts. Expression started 4 days after wounding, reached a maximum at 12 days, and persisted up to 30 days when a scar was formed. After wound closure, most CRBP-1-containing fibroblastic cells underwent apoptosis. We have further investigated CRBP-1 expression in rat fibroblasts cultured from different organs. CRBP-1 was abundant in lung and heart fibroblasts and was detected in decreasing amounts in muscle, tendon, subcutaneous tissue, and granulation tissue fibroblasts. Dermis fibroblasts contained no detectable levels of CRBP-1. All trans retinoic acid and transforming growth factor-beta1 inhibited cell proliferation and increased CRBP-1 expression in fibroblastic populations except dermis fibroblasts. We demonstrate that during granulation tissue formation a subpopulation of fibroblastic cells express CRBP-1 de novo. We also demonstrate that CRBP-1 expression by fibroblasts is regulated in vitro by retinoic acid and transforming growth factor-beta1. Our results suggest that CRBP-1 and possibly retinoic acid play a role in the evolution of granulation tissue. PMID- 9403725 TI - Type 2 helper T-cell predominance and high CD30 expression in systemic sclerosis. AB - The pattern of cytokine production of skin-infiltrating T cells from patients with progressive systemic sclerosis was investigated. Most CD4+ T-cell clones generated from skin biopsy specimens showed a type 2 helper (Th2) cytokine profile (production of interleukin-4, but no interferon (IFN)-gamma). High interleukin-4 but little or no IFN-gamma mRNA expression was found by in situ hybridization in skin perivascular mononuclear cell infiltrates. The immunohistochemical analysis revealed CD30 expression by high numbers of CD4+ T cells in the same specimens. Finally, the great majority of patients with diffuse disease had elevated levels of soluble CD30 in their sera. These data suggest the existence in patients with progressive systemic sclerosis of a predominant activation of Th2-like T cells, which may account for the major alterations (endothelial cell injury, fibrosis, and autoantibody production) occurring in this disease. PMID- 9403726 TI - Prostatic neuroendocrine cells have a unique keratin expression pattern and do not express Bcl-2: cell kinetic features of neuroendocrine cells in the human prostate. AB - We investigated the keratin phenotype and bcl-2 immunoreactivity of neuroendocrine cells in the human prostate to determine whether the postmitotic status of these cells is associated with protection from apoptosis by bcl-2 protein expression and to elucidate the possible cell kinetic relationship between neuroendocrine cells and the other epithelial components of the prostate. Tissue specimens were selected from prostates of 19 patients harboring normal secretory glands (n = 15) and glandular benign prostatic hyperplasia (n = 10). Using a novel sequentially selective destaining immunoenzymatic cytochemical technique we were able to demonstrate the distribution of neuroendocrine cells, keratin markers identifying either basal, luminal, or intermediate cells, and the bcl-2 protein in single sections. Basal cell keratins were expressed in the minority of the neuroendocrine cells. In most of the cells, intermediate and luminal cell keratins were found and bcl-2 was constantly negative. Our findings indicate that neuroendocrine cells and other epithelial cells in the human prostate share a common keratin phenotype and probably originate from a common epithelial precursor. From the absence of bcl-2 we infer that the neuroendocrine cells have no progenitor cell characteristics. PMID- 9403727 TI - Methylation, a major mechanism of p16/CDKN2 gene inactivation in head and neck squamous carcinoma. AB - We studied 11 head and neck squamous carcinoma (HNSC) cell lines and 46 primary tumors for p16 gene status by protein, mRNA, and DNA genetic/epigenetic analyses to determine the incidence, the mechanism(s), and the potential biological significance of its inactivation. Of the 11 cell lines, only 1 showed intact p16 and 10 lacked its protein and mRNA; DNA analysis of these 10 cell lines showed 2 homozygous deletions, 6 methylations at exon 1 and 2, and 2 with no detectable abnormalities. In primary tumors, 16 (34.7%) of the 46 showed detectable p16 protein and mRNA; of these, 12 had no DNA abnormalities and 4 had only exon 2 methylation. Loss of p16 expression was found in three tumors with concurrent mutation at exon 2 and methylation at exon 2 (two) and both 1 and 2 (one). Of the 30 tumors that lacked p16 protein, 27 also lacked mRNA, 1 had detectable p16 mRNA, and 2 failed RT-PCR amplification. Twenty-two of the thirty tumors showed DNA alterations and eight manifested no abnormalities; DNA alterations comprised 6 homozygous deletions, 2 concurrent mutations and methylation of exon 2, and 13 with methylation at exon 1 and exons 1 and 2 (12 with methylation only and 1 with mutation) at exon 1. Except for patients' gender (P = 0.02), no significant correlation between p16 and clinicopathological factors was observed. We conclude that in HNSC 1) intragenic p16 alterations are infrequent events, 2) methylation of exon 1 constitutes a common mechanism in silencing the p16 gene, 3) p16 inactivation may play an important role in the early development and progression of HNSC, and 4) no association between p16 alterations and conventional clinicopathological factors was noted in this cohort. PMID- 9403728 TI - Fibroblast growth factor-2 inhibits endothelial cell apoptosis by Bcl-2-dependent and independent mechanisms. AB - Intact endothelium acts as a sensor and transducer of signals and also provides a nonthrombogenic surface at the blood-vascular wall interface. Hence, mechanisms that maintain the integrity of the endothelium are of interest in physiological and pathological states. In this study we show that apoptosis induced by growth factor and serum deprivation of endothelial cells occurs at all phases of the cell cycle and can be blocked by fibroblast growth factor-2 (FGF-2) independently of its mitogenic activity. As the Bcl-2 family of proteins plays a prominent role in regulating cell survival, we attempted to identify Bcl-2 homologues expressed in endothelial cells. Here we demonstrate that, in addition to the previously identified A1, four other members of the Bcl-2 family, Bcl-2, Mcl-1, Bcl-X(L), and Bax, are expressed in endothelial cells. Of these family members, only Bcl-2 is induced by FGF-2. Overexpression of Bcl-2, using a retroviral vector, protects endothelial cells from serum and growth factor deprivation. There is no difference in FGF-2-induced proliferation between Bcl-2-overexpressing cells and those transduced with the empty retroviral vector. At early time points Bcl-2 is not up-regulated, but FGF-2 still has a protective effect. However, FGF-2 protects only adherent endothelial cells but not those that are cultured in suspension. The early effect of FGF-2 is dependent on tyrosine phosphorylation but not on activation of the MAP kinase pathway. Thus, FGF-2 inhibits endothelial cell apoptosis by Bcl-2-dependent and independent mechanisms. PMID- 9403729 TI - Chronic obstructive pulmonary disease: role of bronchiolar mast cells and macrophages. AB - Chronic obstructive pulmonary disease (COPD) is considered to be caused in part by smoking-induced inflammation, but it is unknown which inflammatory cells within the small airways are associated with the obstruction. We investigated the inflammatory infiltrate in the small airways of 16 current or ex-smokers with COPD (FEV1 < or = 75% predicted) and 15 without COPD (FEV1 > or = 85% predicted) in pneumectomy specimens that were removed for lung cancer. Mast cells, macrophages, neutrophils, eosinophils, T cells, and B cells were identified using immunohistochemistry on formalin-fixed, paraffin-embedded specimens. These cells were quantified in the epithelium and the remainder of the airway wall. The number of mast cells and macrophages in the epithelium, but not in the remainder of the airway wall, was significantly increased in patients with COPD. Neutrophil and T cell numbers did not differ between the groups. Only few B cells and eosinophils were present in both groups. Smoking history, perioperative steroid usage, tumor localization, or reversibility in the FEV1 to salbutamol could not account for the observed differences. We conclude that the number of epithelial mast cells and macrophages is increased in the bronchioli in smokers with airflow limitation, suggesting a role in development of COPD. PMID- 9403731 TI - Accumulation of genetic changes is associated with poor prognosis in grade II astrocytomas. AB - Unexpectedly aggressive clinical course of some grade II astrocytomas is a diagnostic dilemma for routine histopathology. Because increasing tumor malignancy is a consequence of progressive accumulation of chromosomal alterations, we investigated whether aggressive behavior of grade II astrocytomas could be predicted by the number and type of gross chromosomal aberrations. We used comparative genomic hybridization to analyze 11 grade II astrocytomas with typical (good, n = 7) or poor (n = 4) prognosis. The results were also compared with a reference material of 13 grade III-IV astrocytomas and nine established cell lines. We found a median of two aberrations (range 0 to 4) in tumors with good prognosis and of 15.5 changes (range 8 to 28) in tumors with poor prognosis. Chromosomal gains were present in both groups, whereas chromosomal losses were frequent in tumors with poor prognosis (median 9.5, range 3 to 14) but rare in tumors with good prognosis (range 0 to 2). All chromosomal gains were also found in the high-grade astrocytoma group and the majority of them in cell lines. Chromosomal losses in grade II astrocytomas with poor prognosis were very similar to those in grade III-IV astrocytomas and cell lines. We conclude that an early accumulation of genetic changes in grade II astrocytomas is closely associated with poor patient prognosis, suggesting diagnostic use for comparative genomic hybridization in characterization of grade II astrocytomas. PMID- 9403730 TI - Host modifier genes affect mouse autoimmunity induced by the lpr gene. AB - A defect in apoptotic signal transmission through CD95 is an essential genetic mechanism for lymphoproliferation and autoimmunities in lpr or gld mice. However, disease manifestations are largely affected by the host genetic background. To identify and map such host genes modifying lpr gene effect, ie, the lpr modifier (Lprm) genes, 82 MRL/lpr x (MRL/lpr x C3H/lpr) F1 mice were subjected to immunopathological and genetical analyses. High-grade vasculitis and glomerulonephritis among backcross mice were observed in separate groups of mice. Microsatellite analysis revealed that there were two host genes affecting the occurrence of vasculitis, Lprm1 (chromosome 4) and Lprm2 (chromosome 3). A recessive MRL allele at Lprm1 enhanced vasculitis to occur in both sexes, whereas that of Lprm2 inhibited its development selectively in females. Genotype combinations of these two genes explained the severity of vasculitis in crosses of MRL/lpr and C3H/lpr mice and also the vasculitis-prone recombinant inbred strain McH5/lpr. A recessive MRL allele at Lprm3 (chromosome 14) suppressed glomerulonephritis. The weight of the spleen was increased by a recessive MRL allele at Lprm4 (chromosome 5) yielding a logarithm of odds score of 2.02 in a quantitative trait locus analysis. In contrast, the weight of axillary lymph nodes was increased by a recessive MRL allele at a locus on chromosome 2, but its presence was not supported by the quantitative trait locus analysis. The titer of anti-dsDNA autoantibody was controlled by the locus Lprm5 on chromosome 16, which had an logarithm of odds score of 3.41. Possible candidate genes for Lprm genes deduced from their map locations are discussed and compared with the autoimmunity genes reported thus far. In conclusion, autoimmune disease manifestations by the lpr mutation are affected by multiple host genes separately. PMID- 9403732 TI - Human cytotrophoblast differentiation/invasion is abnormal in pre-eclampsia. AB - During human placental development, cytotrophoblast stem cells differentiate and invade the uterus. Simultaneously, the cells modulate their expression of several classes of stage-specific antigens that mark transitions in the differentiation process and play a role in either uterine invasion (integrin cell-extracellular matrix receptors and matrix metalloproteinase-9) or immune interactions (HLA-G). The pregnancy disease pre-eclampsia is associated with shallow cytotrophoblast invasion. Previously we showed, by immunofluorescence localization on placental tissue, that in pre-eclampsia invasive cytotrophoblasts fail to properly modulate their integrin repertoire. This finding suggests possible abnormalities in the differentiation pathway that leads to uterine invasion. Here we used a culture system that supports this differentiation process to compare the differentiative and invasive potential of cytotrophoblasts obtained from control (n = 8, 22 to 38 weeks) and pre-eclamptic (n = 9, 24 to 38 weeks) placentas. In culture, the cells from pre-eclamptic placentas failed to properly modulate alpha1 integrin and matrix metalloproteinase-9 expression at the protein and mRNA levels. Their invasive potential was also greatly reduced. Likewise, the cells failed to up regulate HLA-G protein and mRNA expression. These results suggest that defective cytotrophoblast differentiation/invasion can have significant consequences to the outcome of human pregnancy (ie, development of pre-eclampsia) and that, by the time delivery becomes necessary, the defect is not reversed by removing the cells from the maternal environment. PMID- 9403734 TI - Outcome following cardiopulmonary resuscitation in the pediatric intensive care unit. PMID- 9403733 TI - Duodenal content reflux esophagitis in the rat: an animal model for the ulcer associated cell lineage (UACL)? AB - We have studied the histological changes observed in the mucosa of 10 rats in the region of a esophagojejunostomy to evaluate it as a model for the ulcer associated cell lineage (UACL). In man, the UACL has a distinctive morphology, proliferative organization, and pattern of trefoil peptide localization. We have therefore examined these aspects aided by immunohistochemistry and in situ hybridization to the trefoil peptides TFF1, TFF2, and TFF3. Only TFF2 was studied by immunohistochemistry, whereas the mRNAs for all three peptides were examined by in situ hybridization using 35S-labeled riboprobes. The marker MIB-1 to the Ki67 proliferation-related antigen was used to examine the proliferative organization of UACL-like changes. In all cases, columnar epithelialization of the distal esophagus was seen, and in all, glands with morphological and gene expression attributes of the UACL were identified. TFF3 mRNA localized patchily throughout the UACL, whereas TFF1 mRNA was found in the upper portions of the lineage and TFF2 mRNA and its product in the acini. These lineages showed virtually no intrinsic proliferative activity. These appearances are similar to those seen in early human UACL, and we therefore propose this that this represents the first published animal model of this lineage. PMID- 9403735 TI - Body mass index and the hospitalized patient: a "mean" outcome for the "lean". PMID- 9403736 TI - Optimally managing fluid overload in intensive care. PMID- 9403737 TI - Sedation of the critically ill: goals, plans, and cost-effectiveness. PMID- 9403738 TI - The jury on femoral vein catheterization is still out. PMID- 9403739 TI - Intratracheal pulmonary ventilation versus conventional ventilation in a model of meconium aspiration: searching for a safer and more efficient ventilation modality. PMID- 9403740 TI - Another antiendotoxin strategy to be added to the list. PMID- 9403741 TI - Cardiopulmonary resuscitation in pediatric intensive care units. AB - OBJECTIVE: To determine the effectiveness of cardiopulmonary resuscitation (CPR) in the pediatric intensive care unit (ICU). DESIGN: A nonconcurrent cohort study of consecutive admissions. SETTING: Thirty-two pediatric ICUs. PATIENTS: Consecutive admissions to 32 pediatric ICUs. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Pediatric ICU patients were followed for the occurrence of a cardiopulmonary arrest (external cardiac massage for at least 2 mins). Patients who were in a state of continuous cardiopulmonary arrest on admission, or who never achieved stable vital signs, were excluded from the study. A total of 205 patients, from a sample of 11,165 (1.8%) pediatric admissions, experienced a cardiopulmonary arrest. Overall, 28 (13.7%) patients survived to hospital discharge. Neither mean ages nor age distribution affected survival. Only two diagnostic categories, traumatic illness, and other etiologies, were associated with survival. None of the patients fitting this category survived (p = .0028). The durations of CPR for survivors and nonsurvivors were 22.5 +/- 10.1 and 24.8 +/- 1.9 mins, respectively (p = .015). For CPR durations of <15 mins, 15 to 30 mins, and >30 mins, the survival rates were 18.6%, 12.2%, and 5.6%, respectively (linear trend p = .022). Thirty-five (17.1%) patients had a cardiopulmonary arrest before pediatric ICU admission and another arrest in the pediatric ICU. Only two (5.7%) of these 35 patients survived to discharge. Pediatric ICU survival decreased as the number of pediatric ICU arrests increased. Patients with one arrest (n = 155), two arrests (n = 29), and more than three arrests (n = 21) experienced survival rates of 14%, 14%, and 9.5%, respectively. Severity of illness, as measured by the Pediatric Risk of Mortality III score, was a significant predictor of survival (p < .001). CONCLUSIONS: Pediatric ICU cardiac arrest is an uncommon event. When it does occur, prehospital CPR, duration of resuscitation, traumatic etiology, and severity of illness are important factors associated with survival. PMID- 9403742 TI - Polymyxin-dextran antiendotoxin pretreatment in an ovine model of normotensive sepsis. AB - OBJECTIVE: To test the hypothesis that adult sheep pretreated with polymyxin dextran and then made septic by cecal ligation and perforation would have fewer changes in microvascular integrity and cellular architecture in extrapulmonary organs. DESIGN: Prospective, randomized, double-blind, placebo-controlled animal study. SETTING: An animal research facility in a university-affiliated hospital. SUBJECTS: Mature, male Suffolk sheep (32 to 67 kg). INTERVENTIONS: Animals with chronic indwelling catheters were pretreated with polymyxin B-dextran (6 mg/kg) or placebo (dextran) and an intra-abdominal focus of infection was then produced by cecal ligation and perforation. Treatment (polymyxin B or placebo) was continued every 8 hrs for 48 hrs. MEASUREMENTS AND MAIN RESULTS: Forty-eight hours after randomization, the polymyxin B-dextran group manifested significantly less pyrexia (p = .04), higher mean arterial pressures (p = .02), less variable serum albumin concentrations (p = .05), and a trend toward decreased lactate concentrations (p = .10). Qualitative morphometry and semiquantitative scoring of tissue from gastrocnemius muscle demonstrated that polymyxin B-dextran-treated sheep had significantly increased total capillary (p = .04) and capillary luminal areas (p = .038) and less mitochondrial swelling and damage (p = .03) compared with the placebo sheep. CONCLUSIONS: Pretreatment of sheep in a polymicrobial, peritonitis model of sepsis with polymyxin B-dextran resulted in a significant amelioration of sepsis-induced ultrastructural damage. In placebo-treated control animals, these ultrastructural lesions were associated with a greater severity of sepsis, as measured by the presence of pyrexia, increased lactate concentrations, and less stable blood pressures. These findings justify the investigation of the effects of polymyxin B-dextran in a post onset model of sepsis. PMID- 9403743 TI - Relationship of body mass index to subsequent mortality among seriously ill hospitalized patients. SUPPORT Investigators. The Study to Understand Prognoses and Preferences for Outcome and Risks of Treatments. AB - OBJECTIVE: To determine if body mass Index (BMI = weight [kg]/height [m]2), predictive of mortality in longitudinal epidemiologic studies, was also predictive of mortality in a sample of seriously ill hospitalized subjects. DESIGN: Prospective, multicenter study. SETTING: Five tertiary care medical centers in the United States. PATIENTS: Patients > or = 18 yrs of age who had one of nine illnesses of sufficient severity to anticipate a 6-month mortality rate of 50% were enrolled at five participating sites in the Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments (SUPPORT). INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Patients were asked their current height and weight as part of the demographic data. Stratifying body mass index by percentile rank (< or = 15, 15 to 85, and > or = 85th percentiles), risk ratios for mortality were calculated by Cox Proportional Hazards using the 15th to 85th percentile of body mass index as the reference group while controlling for multiple variables such as prior weight loss, albumin, and Acute Physiology Score. A body mass index in the < or = 15th percentile was associated with an excess risk of mortality (risk ratio = 1.23; p < .001) within 6 months. High body mass index (> or = 85th percentile) was not significantly related to risk of mortality. CONCLUSIONS: Body mass index, a simple anthropometric measure of nutrition employed in community epidemiologic studies, has now been demonstrated to be a predictor of mortality in an acutely ill population of adults at five different tertiary centers. Even when controlling for multiple disease states and physiologic variables and removing from the analysis all patients with significant prior weight loss, a body mass index below the 15th percentile remained a significant and independent predictor of mortality. Examination of patient vs. proxy data did not change the results. Future studies examining variables predictive of mortality should include body mass index, even in acutely ill populations with a poor probability of survival. PMID- 9403744 TI - Protocol-guided diuretic management: comparison of furosemide by continuous infusion and intermittent bolus. AB - OBJECTIVE: To evaluate the safety and relative effectiveness of two diuretic protocols in the intensive care unit (ICU). DESIGN: Prospective, randomized comparative study. PATIENTS: Thirty-three cardiac and medical ICU patients with pulmonary edema or fluid overload for which aggressive diuresis was intended. INTERVENTIONS: Enrolled patients were randomized to fluid management strategies combining fluid restriction and individually adjusted diuretic therapy by either continuous or bolus infusions of furosemide, titrated to achieve negative hourly fluid balance. MEASUREMENTS AND MAIN RESULTS: Cumulative intake minus output (primary endpoint); change in serum creatinine, and length of ICU and hospital stay (secondary endpoints). Diuresis by either protocol was feasible, safe, and effective. The main outcome measures were not significantly different for either group managed with a standardized protocol. CONCLUSIONS: Protocol-guided diuretic management, with individualized titration of dosage to defined physiologic endpoints can be readily and safely implemented in the ICU. Both continuous and bolus diuretic regimens appear equally effective in achieving negative fluid balance. Larger studies with a randomized control arm are needed before these protocols can be recommended as routine practice. PMID- 9403745 TI - Propofol 2% in critically ill patients: effect on lipids. AB - OBJECTIVE: To investigate the concentrations of triglyceride, cholesterol, and high-density lipoprotein during a 50-hr infusion of 2% propofol, starting within 24 hrs of admission to the intensive care unit (ICU). DESIGN: Prospective, clinical study. SETTING: ICU, university hospital. PATIENTS: Thirty adult patients, who were ventilated and expected to be sedated for >2 days, were studied for 50 hrs, beginning at 1800 hrs on the first day of ICU admission. MEASUREMENTS AND MAIN RESULTS: Triglyceride, cholesterol, and high-density lipoprotein were measured at 2000, 0400, and 0800 hrs. Tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, and C-reactive protein were measured at 2000 hrs. Median cholesterol and high-density lipoprotein concentrations were at the low end of the normal range. In seven patients, peak triglyceride concentrations were >3 mmol/L up to a maximum of 4.83 mmol/L. Although there was no statistical difference in lipid concentrations between days 1 and 2, there was an apparent pattern of increasing triglyceride concentrations. There was a correlation between peak triglyceride concentration and total propofol consumption, but there was no correlation between lipids and age, gender, or Acute Physiology and Chronic Health Evaluation II scores. There was a direct correlation between triglyceride and C-reactive protein concentrations, and an inverse correlation between cholesterol and C-reactive protein. Twenty-two patients had evidence of TNF and 11 patients had an IL-6 of >1000 pg/mL, but there was no relationship between concentrations of cytokines and triglycerides in plasma. CONCLUSIONS: Infusion of 2% propofol to critically ill patients over a 50-hr period does not result in a significant increase in triglyceride concentrations. Mean cholesterol and high-density lipoprotein concentrations were low throughout the study period. There was a significant direct correlation between triglyceride and C-reactive protein and an inverse correlation between cholesterol and C-reactive protein, suggesting that the changes in lipids in critically ill patients may be partly attributable to the acute-phase response. PMID- 9403746 TI - Lower extremity deep vein thrombosis: a prospective, randomized, controlled trial in comatose or sedated patients undergoing femoral vein catheterization. AB - OBJECTIVES: To determine the rate of lower extremity deep vein thrombosis after the use of femoral catheters in intensive care unit (ICU) comatose or sedated adults. Results were then compared with results of patients undergoing superior vena cava catheterization. DESIGN: Prospective, randomized, controlled, unblinded study. SETTING: A mixed medical/surgical ICU in a university hospital. PATIENTS: Sixty-one comatose or sedated patients admitted to the ICU who underwent central venous catheterization. INTERVENTIONS: Patients were monitored for signs of thrombotic complications. On catheter removal, a lower-extremity bilateral phlebographic examination was performed in each patient. MEASUREMENTS AND MAIN RESULTS: After randomization, 31 patients underwent femoral vein catheterization and 30 patients underwent superior vena cava catheterization, either by axillary (21 patients) or internal jugular vein (10 patients) cannulation. Single lumen polyurethane catheters were inserted for a mean duration of 7.1 +/- 4.6 (SD) days in the femoral vein group and 9.9 +/- 5.5 days in the superior vena cava group (p = NS). No patient had clinical signs of leg venous thrombosis or pulmonary embolism during the study period. In each patient, lower extremity bilateral phlebography was performed at the time of catheter removal. Leg phlebographies were normal in 18 (60%) patients in the femoral vein group and 26 (84%) patients in the superior vena cava group. Fibrin sleeves which developed around the femoral catheters were seen in seven (23.3%) patients in the femoral vein group and in no patients in the superior vena cava cannulation group. Three patients had femoral vein thrombosis, two (6.6%) patients in the femoral vein group (two nonobstructive thromboses, adherent to the common femoral vein wall) and one (3.0%) patient in the superior vena cava group (nonobstructive thrombosis which developed in the superficial femoral vein) (p = NS). Lower deep extremities thrombosis developed in five (16.7%) patients in the femoral vein group and in five (16%) patients in the superior vena cava group (p = NS). CONCLUSIONS: Femoral vein catheterization with a polyurethane catheter is associated with a lower rate of extremity deep vein thrombosis which is similar to the rate observed after superior vena cannulation in comatose or sedated patients. Femoral vein thrombosis was observed at a rate of 6.6% after femoral vein cannulation and a rate of 3% after superior vena cava cannulation. Given the acceptable rate of this clinically important complication, femoral vein cannulation offers an attractive alternative to insertion via the vena cava in the critically ill. PMID- 9403747 TI - A prospective evaluation of the use of femoral venous catheters in critically ill adults. AB - OBJECTIVE: To determine the rate of complications following the use of femoral catheters in adults. DESIGN: Prospective survey of major and minor complications. SETTING: A mixed medical/surgical intensive care unit (ICU) in a university hospital. PATIENTS: Eighty consecutive patients admitted to the ICU who underwent right femoral venous catheterization over a 13-month period. INTERVENTIONS: Patients were carefully monitored for mechanical, infectious, and thrombotic complications. On catheter removal, a lower extremity bilateral phlebographic examination was performed in each patient. MEASUREMENTS AND MAIN RESULTS: There were 80 polyurethrane catheters inserted for a mean duration of 8.8 +/- 4.4 (SD) days. Catheters were inserted by interns or residents (75%) or by critical care fellows (25%). Minor complications consisted of arterial puncture (15%), local hematoma (4.4%), local bleeding (3%), and local inflammation (5%). After insertion, 17% of catheter tips were in the right atrium, 13% in the abdominal vena cava, 63% in the thoracic inferior vena cava (correct position), and 8% in aberrant abdominal intravascular positions. After repositioning, 80% of catheter tips were in the thoracic inferior vena cava. One patient developed a catheter related bacteremia. Catheter-related sepsis were seen in three (3.7%) patients and catheter colonization in 11 (13.7%) patients. No patient had clinical signs of deep vein thrombosis or pulmonary embolism. Bilateral phlebography was performed in 70 patients at the time of catheter removal and was normal in 45 (64%) patients. Fibrin sleeves were seen in 11 (15.7%) patients. Lower extremity deep vein thrombosis developed in 24 (34%) patients. Six (8.5%) patients had femoral vein thrombosis (common femoral vein in two patients, and superficial femoral vein in four patients). Eighteen (25.7%) patients developed popliteal vein or posterior tibial vein thrombosis that was either bilateral (n = 16) or homolateral (n = 2) to the femoral catheter. CONCLUSIONS: Based on the data from this study, we conclude that femoral vein catheterization with a polyurethane catheter is associated with an 8.5% frequency rate of femoral vein thrombosis. Thrombosis in the popliteal vein or posterior tibial vein is higher (25.7%), but is homolateral to the catheter with only a 2.8% frequency rate. Infectious complications are low and similar to those of other central venous routes. Given the acceptable rate of clinically important complications, femoral venous catheterization offers an attractive alternate site of insertion to the jugular and subclavian veins for central venous access in the critically ill. PMID- 9403748 TI - Sympathetic response during cardiopulmonary bypass: mild versus moderate hypothermia. AB - OBJECTIVE: To determine the sympathetic response during cardiopulmonary bypass at mild (34 degrees C) and moderate (28 degrees C) hypothermia. DESIGN: A randomized study. SETTING: Tertiary university hospital. PATIENTS: Adults undergoing elective coronary artery bypass graft surgery. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Plasma norepinephrine, epinephrine, and neuropeptide Y concentrations were measured. Moderate, but not mild, hypothermic cardiopulmonary bypass evoked a significant sympathetic response with increases in plasma norepinephrine and neuropeptide Y concentrations. A significantly higher incidence of postoperative atrial fibrillation was also observed in the moderate hypothermic compared with the mild hypothermic group. CONCLUSIONS: Our results indicate that the degree of hypothermia significantly influences the sympathetic response during cardiopulmonary bypass. The higher incidence of postoperative atrial fibrillation in the moderate hypothermic group suggests that the enhanced sympathetic response might be one contributing factor in the development of atrial fibrillation. PMID- 9403749 TI - Systemic inflammatory response syndrome and organ dysfunction following gastrointestinal surgery. AB - OBJECTIVES: Progression from systemic inflammatory response syndrome (SIRS) to sepsis, severe sepsis, and septic shock has been demonstrated in a variety of patients. However, the presence of SIRS alone was not helpful in predicting the development of multiple organ dysfunction syndrome (MODS) since SIRS includes many nonprogressive conditions. This study was conducted to investigate the clinical significance of SIRS in postoperative patients. DESIGN: Retrospective study. SETTING: The surgical department of a university hospital. PATIENTS: Two hundred ninety-two consecutive patients who received elective common gastrointestinal surgery (esophagectomy, pancreatoduodenectomy, hepatectomy, gastrectomy, colorectal resection, and laparoscopic cholecystectomy) between 1992 and 1995. INTERVENTIONS: Patients were analyzed for preoperative physiologic status, surgical stress parameters, and postoperative status of SIRS, complications, and end-organ dysfunction. MEASUREMENTS AND MAIN RESULTS: Duration of SIRS or positive criteria's number of SIRS after surgery significantly correlated with surgical stress parameters (blood loss/body weight and operation time) and peak serum C-reactive protein concentrations. SIRS that continued or reappeared after postoperative day 3 was an early sign of postoperative complications. SIRS continuing consecutively for 2 days after postoperative day 3 had a 70.6% positive predictive value and a 92.5% negative predictive value for postoperative complications. Septic complications and prolongation of SIRS were associated with MODS. Five of six patients who met the SIRS criteria for >30 days developed severe MODS, and three of them died. CONCLUSIONS: SIRS is a useful criterion for the recognition of postoperative complications and end-organ dysfunctions. Early recovery from SIRS may arrest the progression of organ dysfunction. PMID- 9403750 TI - Impact of different customization strategies in the performance of a general severity score. AB - OBJECTIVE: To compare the impact of two different customization strategies in the performance of the admission Mortality Probability Model II (MPM II0) using formal statistical assessment. DESIGN: Analysis of the database of a multicenter, multinational, prospective cohort study, EURICUS-1, involving 89 intensive care units (ICUs) from 12 European countries. SETTING: Eighty-nine ICUs from 12 European countries. PATIENTS: Data from 16,060 patients consecutively admitted to 89 ICUs from 12 European countries were collected during a 4-month period. In accordance with original MPM II0 criteria, the following patients were excluded from analysis: patients <18 yrs of age; patients considered readmissions; patients with acute myocardial infarction or burns; and patients in the postoperative period recovering from coronary artery bypass surgery. A total of 10,397 patients were analyzed. INTERVENTIONS: Collection of the data necessary for the calculation of MPM II0 and basic demographic statistics. Vital status at hospital discharge was registered. Two new logistic regression equations were developed to relate MPM II0 to mortality after splitting the database into development and validation samples, the first with the original logit of MPM II0 as an independent variable (first-level customization), and the second with all 15 original variables (second-level customization). Discrimination (area under the receiver operating curve), Hosmer-Lemeshow goodness-of-fit tests H and C, and observed/expected mortality ratios were evaluated in both samples and within relevant subgroups. MEASUREMENTS AND MAIN RESULTS: The discriminative capability of the models was only slightly affected by customization (0.810 vs. 0.803), remaining lower than in the original description of the MPM II0 (0.824). Calibration improved, with Hosmer-Lemeshow goodness-of-fit tests H and C showing a good fit of the models. However, the formal evaluation of discrimination, calibration, and observed/expected mortality ratios across relevant subgroups appeared to be poor in some groups. CONCLUSIONS: In this ICU patient database, second-level customization was more effective than first-level customization in improving the overall goodness-of-fit of MPM II0 and should probably be chosen as the preferential strategy to improve the fit of a model when the sample size is large enough. However, second level customization had only a slight impact on discrimination. Its effects on the uniformity of fit are insufficient to overcome the problems that can arise when the model is applied in populations in which the case-mix is distinct from the population where it was originally developed. PMID- 9403751 TI - Early bloodstream infection after cardiopulmonary bypass: frequency rate, risk factors, and implications. AB - OBJECTIVE: To determine the incidence, predisposing factors, and outcome of early bloodstream infection after cardiopulmonary bypass. DESIGN: A case control study. SETTING: A 54-bed cardiac surgical intensive care in a tertiary referral center. PATIENTS: Patients from a 30-month period with preoperative hospital stay of <48 hrs and subsequent bloodstream infection within 96 hrs of cardiopulmonary bypass were included in a case group. The control group consisted of patients who had cardiac surgery on the same day as the case group. MEASUREMENTS AND MAIN RESULTS: Patient demographics, history of comorbidity, preoperative laboratory testing, details of surgery, transfusion requirement, inotropic infusions, hemodynamics, and arterial blood gases on admission to intensive care were compared in the two groups. Measures of outcome were duration of mechanical ventilation and intensive care stay, serum creatinine on the first postoperative day, highest creatinine and bilirubin concentrations, and hospital mortality. During the study period, 7,928 patients had cardiac surgery. Sixteen (0.2%) patients had early bloodstream infection; the control group consisted of 95 patients. Thirteen of the patients with bloodstream infection had Gram-negative bacilli on blood culture, two had Candida species, and two had Gram-positive bacteria. On multivariate logistic regression analysis, greater prevalence of preoperative pulmonary hypertension (odds ratio 9; 95% confidence interval 2 to 41.8; p = .004), diabetes (odds ratio 4.6; 95% confidence interval 1.4 to 15.8; p = .01), number of blood products transfused (odds ratio 1.09; 95% confidence interval 1.04 to 1.17; p = .005), and infusion of inotropes (odds ratio 4.7; 95% confidence interval 1.3 to 16.4; p = .02) or vasopressors (odds ratio 4.1; 95% confidence interval 1.3 to 15.6; p = .02) were associated with postoperative bloodstream infection. Early bloodstream infection was associated with significantly prolonged duration of mechanical ventilation (117.2 +/- 21.5 vs. 18 +/- 8.8 hrs; p = .0001), intensive care stay (213 +/- 27.5 vs. 53 +/- 11.3 hrs; p < .0001), greater creatinine concentrations on the first postoperative day (1.6 +/- 0.1 vs. 1.2 +/- 0.04 mg/dL; p = .0002), greater maximum creatinine concentration (2.4 +/- 0.2 vs. 1.3 +/- 0.1 mg/dL; p < .0001), and greater maximum bilirubin concentration (4.7 +/- 0.6 vs. 1.3 +/- 0.2 mg/dL; p < .0001) when compared with the control group. Five (32%) of 16 bacteremic patients died vs. none of the 95 control patients (p < .0001). CONCLUSIONS: Early bloodstream infection after cardiac surgery is uncommon and involves predominantly Gram-negative bacteria. The risk factors associated with bloodstream infection were preoperative morbidity and more complex surgery. Bloodstream infection was associated with a significantly adverse impact on outcome after cardiac surgery. PMID- 9403752 TI - Circulating complement proteins in multiple trauma patients--correlation with injury severity, development of sepsis, and outcome. AB - OBJECTIVE: To investigate protein complement 3a (C3a) and protein complement 3 (C3) plasma levels in trauma patients directly after the injury, in relation to the patients' outcome, the development of sepsis, or the injury severity, as determined by either the Polytrauma Score (PTS), the Injury Severity Score (ISS), or the Trauma and Injury Severity Score (TRISS). DESIGN: Prospective study. SETTING: Surgical intensive care unit in a university hospital. PATIENTS: Thirty four patients with multiple trauma. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: C3a and C3 concentrations, as well as the C3a/C3 ratio, were determined at the time of the accident (T0), at the emergency admission (T1), 8 hrs after the accident (T2), and every 8 hrs until day 3, every 12 hrs until day 6, and once daily on days 7 and 8. The C3a plasma concentrations and the C3a/C3 ratios of nonsurvivors were significantly greater at T0 or T1 as compared with those survivors (p = .008 or .033). Patients who developed sepsis had higher C3a plasma levels at the scene of accident than patients without complications. However, these differences did not reach statistical significance (p = .051), although a clear trend was apparent. Patients were grouped according to the severity of injury, as determined by either the PTS, ISS, or TRISS. We found significant differences in the both the mean C3a values and the C3a/C3 ratio among the different groups, during the first 8 hrs after the injury. In addition, a significant correlation was observed between the C3a concentration or the C3a/C3 ratio at T0 to T2 and either the ISS (r2 = .49), PTS (r2 = .22) or the TRISS (r2 = .45), which was similar to correlations between injury severity scores themselves (r2 = .36 to .58). CONCLUSIONS: Complement activation occurs immediately after the injury. The degree of activation is a hallmark for the outcome of a patient. Determination of C3a concentrations, at the scene of the accident, may prove helpful to assess the severity of the injury and to determine the prognosis. The amount of C3a and the C3a/C3 ratio may be useful as additional parameters to the existing trauma scoring systems, such as, PTS, ISS, and TRISS. PMID- 9403753 TI - Use of intratracheal pulmonary ventilation versus conventional ventilation in meconium aspiration syndrome in a newborn pig model. AB - OBJECTIVE: To determine whether intratracheal pulmonary ventilation (ITPV) allows for effective oxygenation and ventilation at lower mean airway pressures and peak inspiratory pressures than conventional ventilation in a piglet model of meconium aspiration syndrome. DESIGN: Prospective, interventional study. SETTING: The animal research laboratory at Children's National Medical Center, Washington, DC. SUBJECTS: Twenty newborn piglets, 2 to 7 days of age, weighing 1.8 to 2.8 kg. INTERVENTION: Animals were anesthetized, paralyzed, intubated, and ventilated. Femoral arterial and venous catheters were inserted; 5 mL/kg of 20% meconium in normal saline was instilled into the endotracheal tube. Animals were randomized to either ITPV or conventional ventilation, and settings were adjusted to maintain ideal blood gases, i.e., pH 7.35 to 7.45, PCO2 40 to 45 torr (5.3 to 6 kPa), PO2 80 to 100 torr (10.7 to 13.3 kPa), and SaO2 > or = 90%. Ventilatory settings were adjusted as needed to a maximum of: FIO2 1.0, peak inspiratory pressure 40 cm H2O, positive end-expiratory pressure 5 cm H2O, and respiratory rate 80 breaths/min. MEASUREMENTS AND MAIN RESULTS: Arterial blood gases were taken every 30 mins for 4 hrs and ventilatory settings were adjusted to maintain optimal blood gases. Heart rate, mean arterial blood pressure, and arterial saturation were monitored continuously. The animals in the ITPV group had significantly lower peak inspiratory pressure at 1, 2, 3, and 4 hrs after meconium instillation (p < .018) and significantly lower mean airway pressure at 2, 3, and 4 hrs after meconium instillation (p < .03). The mean peak inspiratory pressure in the ITPV animals ranged from 17 +/- 2.7 cm H2O at baseline to 16.6 +/ 5.7 cm H2O at 4 hrs compared with 16.5 +/- 2.7 cm H2O at baseline to 31.8 +/- 9.1 cm H2O at 4 hrs in the conventionally ventilated animals (p < .04). The mean airway pressure ranged from 6.3 +/- 1.1 mm Hg at baseline to 6.8 +/- 2.5 mm Hg at 4 hrs in the ITPV group compared with 5.5 +/- 1.2 mm Hg at baseline to 10.7 +/- 3.4 mm Hg at 4 hrs in the conventional ventilation group (p < .03). The lungs of the ITPV animals were less hemorrhagic and had less pathologic evidence of injury than the lungs of the conventionally ventilated animals. CONCLUSIONS: These results indicate that ITPV can be used to effectively ventilate and oxygenate piglets with meconium aspiration syndrome at lower mean airway pressures and peak inspiratory pressures than conventional ventilation. This lower pressure causes less injury to the lungs of the animals. PMID- 9403754 TI - Isolyte S, a physiologic multielectrolyte solution, is preferable to normal saline to wash cell saver salvaged blood: conclusions from a prospective, randomized study in a canine model. AB - OBJECTIVES: The purpose of this study is to compare normal saline with Isolyte S as the wash solutions during high-volume cell saver autologous blood transfusion. Normal saline, the standard wash solution in cell saver autologous blood transfusion, is associated with acid-base and electrolyte derangements. Isolyte S is a physiologic, balanced multielectrolyte crystalloid solution that approximates the electrolyte content of plasma. DESIGN: Open-label, prospective, randomized study. SETTING: Research laboratory in a Department of Veterans Affairs medical center. SUBJECTS: Fourteen mongrel dogs, weighing 22 to 23 kg each. INTERVENTIONS: Fourteen mongrel dogs were prospectively randomized to receive normal saline (n = 7) or Isolyte S (n = 7). Animals were anesthetized, received heparin for anticoagulation, and underwent 18 cycles of cell saver autotransfusion. In each cycle, 125 mL of blood was arterially withdrawn, and washed with either normal saline (mEq/L) (sodium 154, chloride 154) or Isolyte S (mEq/L) (sodium 141, potassium 5, magnesium 3, chloride 98, phosphate 1, acetate 28, and gluconate 23). The washed blood was retransfused. MEASUREMENTS AND MAIN RESULTS: Acid-base and electrolyte analyses were performed throughout the study on the systemic blood of each group and compared. By the end of the study, the Isolyte S group had a normal pH and an increased bicarbonate concentration (mEq/L: normal values 24 to 32; normal saline 9.0 +/- 1.9 vs. Isolyte S 13.2 +/- 3.0 [p < .01]) and an increased magnesium concentration (mg/dL: normal values 1.6 to 2.4; normal saline 1.6 +/- 0.2 vs. Isolyte S 2.2 +/- 0.2 [p < .0001]). Additionally, the Isolyte S group had a lower chloride concentration (mEq/L: normal values 95 to 110; normal saline 130 +/- 9 vs. Isolyte S 117 +/- 7 [p < .02]) and a lower potassium concentration (mEq/L: normal values 3.5 to 5.0; normal saline 4.4 +/- 0.5 vs. Isolyte S 3.7 +/- 0.3 [p < .01]). There were no significant differences between normal saline or Isolyte S in the values of PCO2, lactic acid, sodium, total and ionized calcium, inorganic phosphorus, total protein, albumin, hemoglobin, and hematocrit. CONCLUSIONS: Fewer systemic acid base and electrolyte derangements were observed when blood was washed with Isolyte S. Differences between the normal saline and Isolyte S groups are ascribed primarily to the constituents of the wash solution. We conclude that Isolyte S, a physiologic, balanced, multielectrolyte solution, should be considered as the wash solution in high-volume autologous cell saver blood processing and transfusion. PMID- 9403755 TI - Experimental investigation of battery-induced esophageal burn injury in rabbits. AB - OBJECTIVE: In recent years, small high-performance batteries have become very popular. With this increasing miniaturization of batteries, clinicians have noted an increasing frequency rate of esophageal injury due to battery ingestion by infants. The situation is associated with severe injury to the esophagus due to the electrical current produced, particularly in the case of high-performance batteries producing high currents. The pathophysiologic features and complications of esophageal battery burns have not been thoroughly investigated. Our study intended to investigate the pathophysiologic features and complications of esophageal battery burn. DESIGN: Open, randomized, controlled study. SETTING: Experimental animal laboratory in a university hospital. SUBJECTS: Male adult mixed-breed rabbits, 22 wks old and weighing 3 to 3.5 kg. INTERVENTIONS: The experimental rabbit model of esophageal injury due to battery ingestion described herein was designed to study not only the direct influence of contact with the battery but also damage to neighboring tissues and the biochemical and pathologic mechanisms of injury. We investigated the relationship between the direction of the inserted battery and the mechanism underlying these complications. Esophageal burn injury was created by placing a 3-V battery into the esophagus for 9 hrs. MEASUREMENTS AND MAIN RESULTS: The cathode side of the esophagus became increasingly alkaline, while the anode side was acidic. Low-voltage battery burns are likely to be due to secondary chemical reactions caused by the electric current because of acid generated at the anode and alkali at the cathode using a micro pH meter. Injury was significantly more severe on the alkaline side when a battery was placed with its cathode directed toward the trachea. Alkaline complications affecting neighboring tissues were more severe than acid complications. These results indicate that as well as the esophageal mucosa itself being injured, deleterious effects are exerted on surrounding tissues, the severity of which vary depending on the orientation and duration of the battery being lodged in the esophagus. CONCLUSIONS: The direction of the battery cathode, which produces alkali, is important in determining the severity of complications. Based on our investigation of the underlying mechanisms of these complications, we advocate the establishment of treatment guidelines for battery swallowing accidents. PMID- 9403756 TI - Perflubron decreases inflammatory cytokine production by human alveolar macrophages. AB - OBJECTIVE: To determine whether inflammatory cytokine production by stimulated human alveolar macrophages is affected by perflubron exposure. DESIGN: Controlled laboratory investigation of alveolar macrophage function in vitro. SETTING: Research laboratory. SUBJECTS: Cultured alveolar macrophages obtained by bronchoalveolar lavage from eleven normal volunteers. INTERVENTIONS: Endotoxin stimulated alveolar macrophages were treated with perflubron. MEASUREMENTS AND MAIN RESULTS: Alveolar macrophages were stimulated for 1 hr with lipopolysaccharide and then treated with perflubron for 23 hrs. Cell-free supernatants were collected and cytokines were assayed by enzyme-linked immunosorbent assay. Tumor necrosis factor-alpha, interleukin-1, and interleukin 6 were stimulated by lipopolysaccharide (endotoxin) and all of these cytokines were significantly (p < .05) inhibited by perflubron. Cell viability was not affected by perflubron. Basal cytokine concentrations from unstimulated alveolar macrophages were not altered by perflubron. CONCLUSIONS: Exposure of stimulated human alveolar macrophages to perflubron in vitro decreases cytokine production. This observation suggests that perflubron may have anti-inflammatory activity. PMID- 9403757 TI - Importance of textual data in multimodality monitoring. AB - OBJECTIVES: The use of multimodality monitoring of patients in the intensive care unit (ICU) and the subsequent collection and analysis of such data are increasing. The aim of this work was to assess the importance of recording complementary textual data referring to patient care maneuvers, calibrations, and other incidents, in addition to the raw numerical values. DESIGN: A retrospective analysis of multimodality monitoring data, which included comments entered concurrently at the bedside, collected from head-injured patients admitted to an ICU. PATIENTS: One hundred forty-seven patients with a postresuscitation Glasgow Coma Scale score of < or = 12 were monitored for a total of nearly 1 million minutes on up to eight commonly used channels. MEASUREMENTS AND MAIN RESULTS: Approximately 13,000 comments were added to the raw data at the time of collection. The data were subsequently validated using these comments as indicators of artifactual values. The comments were classified into a surprisingly small number of important categories, with the most frequent referring to monitor calibrations and regular ICU care maneuvers. The difference between validated and unvalidated data on the quantity of secondary insult observed was in some cases nearly 50%. CONCLUSIONS: This work demonstrates that such textual information should be recorded concurrently with the raw monitoring values to ensure proper interpretation of the data in any retrospective analysis. Furthermore, it also suggests that a small number of prespecified categories could be used in the on-line validation of such data. PMID- 9403758 TI - Comparison of two different pulse oximeters in monitoring preterm infants. AB - OBJECTIVE: The aim of the study was to test the reliability and variation in the readings of two widely used pulse oximeters in preterm infants. DESIGN: Two different pulse oximeters and a transcutaneous PO2 monitor were used to record the data continuously on a cotside computer database. PATIENTS: Sixteen preterm infants were studied in the Neonatal Unit, Simpson Memorial Maternity Pavilion, Edinburgh, UK. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Approximately one fifth of the time, the pulse oximeter readings could be established as artifactual. Study of the remaining four fifths of the data showed that, on average, the Nellcor pulse oximeter recorded saturation percentages 2.2% higher than the Ohmeda oximeter. CONCLUSIONS: We recommend that all neonatal units adopt a policy of using different saturation alarm limits for these two instruments. We further recommend that other pulse oximeters be tested by a methodology similar to the one we present in this paper, before their use in monitoring oxygenation in preterm infants. PMID- 9403759 TI - Nonradiographic assessment of enteral feeding tube position. AB - OBJECTIVE: To determine whether a clinical, nonradiographic criterion can be used to predict when the tip of a blindly placed feeding tube is in the small intestine. DESIGN: Prospective sample. SETTING: Pediatric intensive care unit at a tertiary care children's hospital. PATIENTS: Critically ill children requiring transpyloric feeding. INTERVENTIONS: The small bowel was intubated, using a blind, bedside transpyloric feeding tube placement protocol. The feeding tube was considered to be in the small bowel when <2 mL of a 10- mL aliquot of insufflated air could be aspirated from the feeding tube. This clinical criterion was confirmed with an abdominal radiograph. MEASUREMENTS AND MAIN RESULTS: Patient age ranged from 1 month to 19 yrs (median 6 months). Weight ranged from 2.2 to 60 kg (median 4.9). Median time to feeding tube placement was 10 mins (range 5 to 60). Eighty-nine percent of the patients were mechanically ventilated, while 28% of these patients were pharmacologically paralyzed. Seventy-five feeding tubes were inserted. There were no known complications. Ninety-nine (74/75) percent of the feeding tubes were positioned in the small bowel. The inability to aspirate insufflated air correctly predicted small bowel intubation with 99% certainty (Sequential Probability Ratio Test, p = .05 and power = .80). This test incorrectly predicted the position of only one feeding tube, the 26th, which was in the stomach. Of the 74 feeding tubes positioned in the small bowel, 13 feeding tubes were in the duodenum and 61 were in the jejunum. CONCLUSIONS: The inability to aspirate insufflated air confirms the transpyloric position of a feeding tube. Other clinical criteria did not successfully predict small bowel intubation. Use of this single test may obviate confirmatory abdominal radiographs in carefully selected patients and may lead to more cost-effective and timely initiation of enteral feedings. PMID- 9403760 TI - Show me the evidence: a critical appraisal of the Pulmonary Artery Catheter Consensus Conference and other musings on how critical care practitioners need to improve the way we conduct business. PMID- 9403761 TI - Pulmonary Artery Catheter Consensus Conference: the first step. PMID- 9403762 TI - Inhaled nitric oxide and pulmonary edema formation. PMID- 9403763 TI - Lactic acidosis and cardiopulmonary bypass. PMID- 9403764 TI - Effect of unilateral once or twice daily milking of cows on milk yield and udder characteristics in early and late lactation. AB - Twelve multiparous British Friesian cows in early (40 +/- 23 d in milk; n = 6) or late (216 +/- 17 d in milk; n = 6) lactation were used to study the effects of milking frequency on yield, udder volume and milk storage within the udder. After a 2 week control period of twice daily milking, diagonally opposed udder halves within a cow were milked once or twice daily for 3 weeks. Milk yield was 28-38% lower from the halves that were milked once daily than from halves that were milked twice daily. The loss of milk yield, expressed as a decrease in the relative milk yield quotient (an index that accounts for pretreatment differences), was greater for cows in early than in late lactation (0.59 v. 0.68). Empty udder-half volume was not decreased by once daily milking, suggesting that no cell loss occurred. Instead, once daily milking reduced the secretion efficiency (units of milk per unit of empty udder-half volume) by 46 and 27% respectively in early and late lactation; thus, at least part of the loss was due to reduced metabolic activity of the mammary epithelium. There were positive correlations between the relative milk yield quotient and the proportion (r = 0.804) or volume (r = 0.644) of cisternal milk in the glands that were milked once daily. These results confirm that, during extended milking intervals, milk loss was smallest for cows that stored a larger proportion of milk in the gland cistern. PMID- 9403765 TI - Relationship between degree of casein hydrolysis and phosphopeptide release. AB - In an approach to develop a commercial-scale process for the production of casein phosphopeptides containing the cluster sequence-SerP-SerP-SerP-Glu-Glu-, we have studied the relationship between casein hydrolysis and phosphopeptide release. The degrees of hydrolysis (DH) of casein using Novo trypsin PTN 3.0 S and pancreatin 4NF independently, at enzyme to substrate (E:S) ratios of 1:50-1:1600 (by weight), were determined using the pH-stat method. Casein phosphopeptides (CPP) were selectively precipitated using Ca2+ and ethanol from the acid clarified hydrolysates. The precipitates were analysed by high performance capillary electrophoresis to calculate individual phosphopeptide yields based on extinction coefficients of the purified peptides. Individual peptides were purified by reversed-phase HPLC and anion-exchange FPLC and characterized by MALDI-TOF mass spectrometry and amino acid sequence analysis. For both enzymes, lowering the E:S ratio resulted in reductions in the DH and the release of the CPP, and an increase in peptide chain length. The longer chain length offset the reduction in release such that the gravimetric yields of CPP preparations remained relatively constant. For Novo trypsin the highest yields of the major cluster peptides (beta-casein(CN)f(1-25), alpha s1-CNf(59-79), alpha s2-CNf(1 21), alpha s2-CNf(46-70) and related peptides) in the selective precipitates were obtained at a casein DH of 17%. At lower DH values (9-15%), there was a decrease in yield of the peptides derived from alpha s1-CN and alpha s2-CN while the yield of the beta-CN-derived cluster peptides remained relatively constant. The CPP produced using pancreatin were found to be truncated at all E:S ratios, relative to the tryptic CPP, owing to higher levels of chymotryptic and carboxypeptidase activities in pancreatin. The highest yields of the truncated forms of the major cluster peptides using pancreatin were obtained at a casein DH of 19-23%. PMID- 9403766 TI - Study of the polymorphism of caprine milk caseins by capillary electrophoresis. AB - Polymorphism of caprine milk proteins was studied by capillary electrophoresis. Identification of casein (CN) fractions was effected by using isolated fractions from cation-exchange fast protein liquid chromatography. Genetic polymorphisms in caprine alpha s2-CN, alpha s1-CN, beta-CN and kappa-CN have been determined. kappa-CN A and B, beta-CN A and null, alpha s2-CN A, B and C, alpha s1-CN A, B, C and null, and other forms with intermediate and low alpha s1-CN content have been identified. The capillary electrophoresis method made it possible to analyse whole caprine milk using simple sample preparation and was rapid, automated and suitable for phenotyping studies. This method may also permit the quantitative study of different protein fractions. PMID- 9403767 TI - Study of the polymorphism of ovine alpha s1- and alpha s2-caseins by capillary electrophoresis. AB - Polymorphism of ovine alpha s-caseins was studied by capillary electrophoresis at pH 3.0 +/- 0.1. Individual caseins (CN) were selected according to their genetic variants, as determined by PAGE and isoelectric focusing. The ovine caseins, containing different genetic variants of alpha s1-CN and alpha s2-CN, were fractionated by cation-exchange FPLC. The alpha s1-CN variants A, B and C, and the fast moving alpha s2-CN variant were identified by a capillary electrophoresis method. The fast moving alpha s2-CN variant, so-called for its behaviour in PAGE, also had a faster electrophoretic mobility than the common alpha s2-CN when analysed by the present technique. The capillary electrophoresis method gave excellent, rapid, automated separation of alpha s1-CN and alpha s2-CN variants and was suitable for screening studies. PMID- 9403768 TI - Use of a single-strand conformation polymorphism analysis to perform a simple genotyping of bovine kappa-casein A and B variants. AB - We propose an alternative method for casein genotyping, generally carried out using polymerase chain reaction followed by restriction fragment length polymorphism analysis. Application of the single-strand conformation polymorphism technique detects nucleotide changes in the fragment amplified by means of polymerase chain reaction and thus avoids the use of restriction enzymes. A 453 bp fragment from exon IV of kappa-casein has been amplified. The two variants (A and B), found with the highest frequencies in most bovine breeds and included in some dairy cattle selection schemes, can be discriminated using single-strand conformation polymorphism analysis of heat denatured fragments in acrylamide-bis acrylamide (100:1) gels followed by silver staining. kappa-Casein genotyping is therefore simplified, although variants A and E on the one hand, and B and C on the other, are not distinguishable with this technique. PMID- 9403769 TI - Diarrhoeal toxin production at low temperature by selected strains of Bacillus cereus. AB - The growth of four Bacillus cereus strains producing diarrhoeal toxin at 32 degrees C (F4433/73 and 29.155, isolated on the occasion of foodborne outbreaks, and F4581/76L and F4581/76R, two variants of a clinical strain), a weakly toxigenic strain isolated in routine analysis of food (3505M) and an emetic isolate (F3502/73) was investigated at low temperature. Biomass was determined by protein assay. Generation times were: for strain F3502/73, which grew at > or = 12 degrees C, 8.71 h (at 12 degrees C); for other strains, which grew at > or = 10 degrees C, 10.2 to approximately 18.9 h (at 10 degrees C). Toxin production during growth was evaluated by a commercial kit (Oxoid) and by a toxicity test on Chinese hamster ovary cells. Strains F4433/73 and F4581/76, secreting high levels of diarrhoeal toxin during the exponential phase at 32 degrees C, produced high levels of toxicity at 10 degrees C until the stationary phase. Strain 29.155 had decreased toxin production at 10 degrees C. Toxicities for cellular extracts remained low when compared with culture filtrates. A correlation was found between the toxicity values given by the two detection methods tested, and the suitability of both methods for the detection of potential poisoning isolates is discussed. PMID- 9403770 TI - Phosphopeptides from Grana Padano cheese: nature, origin and changes during ripening. AB - Casein phosphopeptides (CPP) which develop in Grana Padano cheese at different ages were isolated by precipitation with Ba2+ and analysed by HPLC. Profiles were complex throughout the period between 4 and 38 months. CPP in a cheese sample 14 months old were identified by a combination of fast atom bombardment-mass spectrometry and Edman degradation. They were found to consist of a mixture of components derived from three parent peptides, beta-CNf(7-28)4P, alpha s1-CNf(61 79)4P and alpha s2-CNf(7-21)4P. In total, 45 phosphopeptides were identified: 24 from beta-CN, 16 from alpha s1-CN and 5 from alpha s2-CN. The presence of aminopeptidase activity during cheese ripening was deduced from the presence of a number of CPP of different lengths with the loss of one or more residues from the N-terminus. The longest had C-terminal lysine and seemed to be progressively hydrolysed by carboxypeptidases A and B to shorter peptides. CPP in cheese appeared to be shortened plasmin-mediated products. Moreover, those most resistant to further hydrolysis contained at least three closely located phosphoserine residues. The anticariogenic activity of CPP is also discussed. PMID- 9403771 TI - Effect of feed availability on post-milking standing time in dairy cows. PMID- 9403772 TI - Effect of altering the daily herbage allowance in mid lactation on the composition and processing characteristics of bovine milk. PMID- 9403773 TI - Naloxone and adrenergic blocking agents fail to abolish central inhibition of milk ejection in cows. PMID- 9403774 TI - Cortisol, parathyroid hormone-related protein and the onset of calcium secretion by the mammary gland of the goat. PMID- 9403775 TI - Genetic polymorphism of plasminogen in dairy cattle. PMID- 9403776 TI - Comparison of five incubation systems for rat liver slices using functional and viability parameters. AB - Precision-cut liver slices are presently used for various research objects, e.g. to study metabolism, transport, and toxicity of xenobiotics. Various incubation systems are presently employed, but a systematic comparison between these incubation systems with respect to preservation of slice function has not been performed yet. Therefore, we started a comparative study to evaluate five of these systems: the shaken flask (an Erlenmeyer in a shaking water bath), the stirred-well (24-well culture plate equipped with grids and magnetic stirrers), rocker platform (6-well culture plate with Netwell insert rocked on a platform), the roller system (dynamic organ culture rolled on an insert in a glass vial), and the 6-well shaker (6-well culture plate in a shaking water bath). The liver slices were incubated in these incubation systems for 0.5, 1.5, and 24.5 h and subsequently subjected to viability and metabolic function tests. The viability of the incubated liver slices was evaluated by: potassium content, MTT assay, energy charge, histomorphology, and LDH leakage. Their metabolic functions were studied by determination of the metabolism of lidocaine, testosterone, and antipyrine. Up to 1.5 h of incubation all five incubation systems gave similar results with respect to viability and metabolic function of the liver slices. However, after 24 h, the shaken flask, the rocker platform, and the 6-well shaker incubation systems appeared to be superior to the stirred well and the roller incubation systems. PMID- 9403777 TI - Development of a monoclonal antibody to 6 beta-hydroxycortisol and its application in an enzyme-linked immunosorbent assay (ELISA) for 6 beta hydroxycortisol in urine. AB - A novel monoclonal antibody to 6 beta-hydroxycortisol (6 beta-OHC) was generated and incorporated into an antigen-coated indirect enzyme-linked immunosorbent assay (ELISA) using 6 beta-OHC-protein conjugate as the steroid-coating antigen. The monoclonal antibody is specific to 6 beta-OHC and 6 beta-OHC-3 carboxymethyloxime. Cross-reactivity with other structurally related steroids such as cortisol, cortisone, and 6 beta-hydroxycortisone was less than 10%. Two different clones (clone 5C1 and 19F) of the monoclonal anti-6 beta-OHC antibody have been developed, each with slightly different sensitivity and specificity. The sensitivity of the MAb clones was not significantly improved when compared to the rabbit polyclonal antibodies in this study, but still within the accepted detection limit for 6 beta-OHC in both human and laboratory animals. The assay had a detection limit of 200 ng/ml, an intraassay variation of 6.4% and an interassay variation of 7.3%. The application of the anti-6 beta-OHC-MAb-based ELISA was tested by measuring the urinary output of 6 beta-OHC in human before and after enzyme induction by rifampicin treatment. The mean 24-h urine output of 6 beta-OHC in human subjects was 485 +/- 100 micrograms and 1478 +/- 281 micrograms before and after rifampicin administration, respectively. In conclusion, the monoclonal anti-6 beta-OHC antibody developed in this study has the required specificity and sensitivity as an alternative method for measuring urinary 6 beta-OHC in the detection of enzyme induction or enzyme inhibition of CYP3A in humans and laboratory animals. PMID- 9403778 TI - On the reliability of affinity and efficacy estimates obtained by direct operational model fitting of agonist concentration-effect curves following irreversible receptor inactivation. AB - Recently, Zernig and colleagues (1996) (J Pharmacol Toxicol Meth 35: 223-237) suggested that for the estimation of agonist affinity and efficacy, the method of simultaneously fitting of concentration-effect curves from control and irreversible antagonist-treated tissues to the operational model of agonism is superior to other analytical approaches. In the present study, we have evaluated the limitations of this simultaneous curve fitting method. Simulation studies showed that this method can be only employed with confidence when the upper asymptotes of the control curves display minimal variation between tissues, which makes its practical utility rather limited. The unreliability of the simultaneous fitting procedure was further underscored with the analysis of experimental data obtained from the interaction between noradrenaline and phenoxybenzamine in rat isolated aorta. The lack of robustness of the parameter estimates showed that under standard experimental conditions the outcomes of simultaneous model fitting are highly dependent on between-tissue variations of the upper asymptotes of the control curves and, therefore, may be unreliable. Therefore, whenever possible, a multiple curve design should be adopted, in which control and treated curves are obtained in one tissue and provide enough information for an independent estimation of affinity and efficacy that is free of intertissue differences. PMID- 9403779 TI - Tissue water content in rats measured by desiccation. AB - Tissue water content was determined by desiccation to constant weight at 40 degrees-50 degrees C in 14 tissues from two groups of rats weighing 200-250 and 270-430 g, respectively. The water content (mean +/- SE; ml/g) was highest in testes (0.861 +/- 0.002) and lowest in adipose (0.183 +/- 0.017) followed by bone (0.446 +/- 0.017) and skin (0.651 +/- 0.007). The average water content in the remaining tissues was 0.763 (+/- 0.003). Upon correction for the water content of residual tissue blood, significant difference between the uncorrected and corrected tissue water was observed for spleen, lungs, kidneys, heart, liver, and brain. Tissue water was independent of body weight, and was the same for right and left kidneys as well as testes and bone. Whereas the position of the muscle (back, abdomen, hindlimb) and adipose tissue (perirenal and subcutaneous) had no influence on water content, for skin, a slight difference was found between back and abdomen. In general, the current results are in agreement with composite literature values, but provide in one study data for all tissues used in the development of physiologically based pharmacokinetic models of rat. PMID- 9403780 TI - Determination of nitric oxide generation in mammalian neurons using dichlorofluorescin diacetate and flow cytometry. AB - A method for the rapid detection of intracellular nitric oxide (NO) generation in dissociated cerebellar granule cells using dichlorofluorescin (DCFH) and flow cytometry was developed. DCFH can be oxidized specifically by NO and this was assessed by 1) the use of SIN-1 (10 nM-100 microM), an NO donor, that induced a concentration-dependent increase in dichlorofluorescein (DCF) fluorescence and 2) the use of hemoglobin (10 microM), an NO-scavenger, that totally inhibited the increase of fluorescence induced by SIN-1 (10 microM). This assay was used to determine the ability to kainate to stimulate NO production in dissociated cerebellar granule cells. Kainate (1 microM-10 mM) induced an increase in DCF fluorescence that was partially reduced by NG-nitro-L-arginine (1 nM-10 microM), a nitric oxide synthase inhibitor (61.9% +/- 9.1), or hemoglobin (10 microM) (55.0% +/- 4.1). The method described allows evaluation of the oxidation of DCFH to produce DCF as a parameter for measuring intracellular NO generation. The extent of DCFH oxidation by NO and ROS can be determined by using NO scavengers or NO synthase inhibitors. PMID- 9403781 TI - Pharmacokinetic-pharmacodynamic modelling of the EEG effect of alfentanil in rats. AB - The purpose of the present investigation was to develop a methodology for quantification of the concentration-pharmacological effect relationship of alfentanil in individual rats by using effect parameters derived from quantitative EEG analysis. In particular, the role of the opioid-induced side effects of proconvulsant activity, hypothermia, and respiratory depression was investigated. Amplitudes in the 0.5- to 4.5-Hz frequency band of the EEG power spectrum were used as a descriptor of the effect. The pharmacokinetics and pharmacodynamics of alfentanil were determined after intravenous administration of 2000 micrograms/kg during 20 min. The administration of alfentanil induced paroxysmal seizure patterns in the EEG which made meaningful analysis of the EEG effect impossible. A constant infusion of midazolam (5.5 mg/kg/hr) prevented alfentanil-induced seizures. When no precautions were taken to control body temperature, analysis of the concentration-EEG effect relationship was complicated by proteresis due to alfentanil-induced hypothermia. This proteresis disappeared when body temperature was stabilized at 37.5 degrees-38.5 degrees C with isothermal pads. Alfentanil-induced respiratory depression was managed successfully by artificial ventilation. Adequateness of artificial ventilation was ascertained by monitoring of arterial pO2, pCO2, and pH. When these opioid side effects were controlled, the pharmacokinetics were most adequately described by a biexponential function. The values of the pharmacokinetic parameters were (mean +/- SE, n = 7); clearance = 53 +/- 6 ml.min/kg, volume of distribution = 1.19 +/- 0.19 l/kg and terminal half-life = 24.5 +/- 2.3 min. By pharmacokinetic pharmacodynamic modelling, the individual concentration-effect relationships of alfentanil were derived, which were successfully characterized by the sigmoidal Emax pharmacodynamic model. The values of the pharmacodynamic parameters were (mean +/- SE, n = 7): E0 = 57 +/- 4 microV, Emax = 95 +/- 17 microV, EC50 = 202 +/- 55 ng/ml and Hill factor = 1.53 +/- 0.20. No delay was observed between alfentanil concentration and effect. The results of the present study show that when side effects are controlled adequately, the concentration-EEG effect relationship of alfentanil can be characterized in individual rats using amplitudes in the 0.5- to 4.5-Hz frequency band of the EEG as a measure of the pharmacological response. The described methodology can be very useful for detailed studies into the pharmacodynamics of (new) synthetic opioids in vivo. PMID- 9403782 TI - Audible and ultrasonic vocalization elicited by a nociceptive stimulus in rat: relationship with respiration. AB - Brief electrical pulses applied to a rat's tail elicit complex vocal responses including audible ("peeps," "chatters") and ultrasonic components. These responses, particularly the two first peeps which have been shown to be triggered by A delta- and C-fibers, could provide a useful tool in pain studies. In the present study, we aimed to optimize this test by investigating the influence of respiration on the vocal responses. The following results were obtained: 1) As expected, the vocalization periods were concomitant with expiration; 2) The phase of the respiratory cycle at the onset of stimulation did not modify the mean intensities of the peeps; 3) The lung volume at the onset of stimulation significantly influenced the intensity and duration of the first peep and the latency of the second peep. Taking account of respiratory parameters in pain tests based on a quantified analysis of vocal responses could improve their sensibility by reducing variability and their specificity by detecting confounding factors such as effects of drugs on respiratory centres or on motor function. PMID- 9403783 TI - Developmental factors modify stress ulcer incidence in a stress-susceptible rat strain. AB - Our multifactor theory of stress ulcer assumes that environmental factors that operate during early growth stages influence the elaboration of stress ulcer in adult rats. The theory would predict that rats exposed to either neonatal handling, or raised in a stimulus enriched environment, would reveal differences in stress ulcer susceptibility. In study 1, some Wistar rats and ulcer susceptible Wistar Kyoto (WKY) rats were handled daily from birth to day 21, whereas other rats from each strain were not disturbed. In study 2, Wistar and WKY rats were raised (3 months) in a large stimulus-dense enriched environment, whereas other rats from each strain were raised in standard rat cages where visual and auditory stimuli were minimized. At 3 months all rats were observed in the open field test (OFT), a test of emotionality, as well as the Porsolt forced swim test (FST), a test of behavioral depression, and subsequently exposed to the ulcerogenic water restraint procedure. Neonatal handling produced results suggesting increased wall climbing activity in the FST, reduced response latency in the OFT, increased body weight and reduced ulcer severity, but these differences were not significant. Rearing in an enriched environment produced similar results but these difference were more pronounced and significant in the Wistar rats as compared to the WKY rats. Thus early environmental manipulations can influence adult behavior and the elaboration of stress ulcer disease, but the impact of these manipulations is less salient in an organism with an endogenous susceptibility to the disease. PMID- 9403784 TI - Pentadecapeptide BPC 157 positively affects both non-steroidal anti-inflammatory agent-induced gastrointestinal lesions and adjuvant arthritis in rats. AB - Besides a superior protection of the pentadecapeptide BPC 157 (an essential fragment of an organoprotective gastric juice peptide BPC) against different gastrointestinal and liver lesions, an acute anti-inflammatory and analgetic activity was also noted. Consequently, its effect on chronic inflammation lesions, such as adjuvant arthritis, and non-steroidal anti-inflammatory agents (NSAIAs)-induced gastrointestinal lesions was simultaneously studied in rats. In gastrointestinal lesions (indomethacin (30 mg/kg s.c.), aspirin (400 mg/kg i.g.) and diclofenac (125 mg/kg i.p.) studies, BPC 157 (10 micrograms or 10 ng/kg i.p.) was regularly given simultaneously and/or 1 h prior to drug application (indomethacin). In the adjuvant arthritis (tail-application of 0.2 mL of Freund's adjuvant) studies (14 days, 30 days, 1 year) BPC 157 (10 micrograms or 10 ng/kg i.p.), it was given as a single application (at 1 h either before or following the application of Freund's adjuvant) or in a once daily regimen (0-14th day, 14 30th day, 14th day-1 year). Given with the investigated NSAIAs, BPC 157 consistently reduced the otherwise prominent lesions in the stomach of the control rats, as well as the lesions in the small intestine in the indomethacin groups. In the adjuvant arthritis studies, the lesion's development seems to be considerably reduced after single pentadecapeptide medication, and even more attenuated in rats daily treated with BPC 157. As a therapy of already established adjuvant arthritis, its salutary effect consistently appeared already after 2 weeks of medication and it could be clearly seen also after 1 year of application. Taking together all these results, the data likely point to a special anti-inflammatory and mucosal integrity protective effect. PMID- 9403785 TI - Capsaicin-sensitive mechanisms in the modulation of rat colonic vascular permeability under physiological and pathological conditions. AB - Inflammatory bowel disease (IBD) causes a prolonged life-quality reduction of patients and high costs for health services. The aim of this study was to explore the possible involvement of peptidergic capsaicin-sensitive afferent nerves (CSN) in the pathogenesis of IBD. For the defunctionalization of colonic CSN, the lower part of the colon (1-4 cm from the anus) was exposed through a midline laparotomy and small pieces of gelfoam moistened with a solution of capsaicin (1%, 100 microL) was applied onto the serosal surface for 30 min in male Wistar rats. Colonic vascular permeability was assessed by measuring the extravasation of [125I] human serum albumin (2 microCi/kg, i.v., 2 h prior to killing). Two months after capsaicin treatment a significant increase in albumin extravasation was found in the lower (P < 0.005), but not in the upper (5-8 cm from the anus) part of the colon as compared to the sham-operated control. Intrarectal (8 cm from anus) administration of trinitrobenzene-sulphonic acid (TNBS; 30 mg/rat) induced similar plasma leakage in the lower and upper colon of control (CSN-intact) rats (P < 0.001) 1 week later. TNBS + ethanol (50%) produced further extravasation throughout the colon (P < 0.001) of CSN-intact animals. In the lower colon of capsaicin-pretreated rats TNBS-alone provoked an increase in plasma extravasation (P < 0.001) similar to that caused by TNBS + ethanol in CSN-intact rats. In the upper colon there was no difference in the effect of TNBS-alone on plasma leakage between control (CSN-intact) and CSN-depleted rats. The results suggest that capsaicin-sensitive nerves may play a significant protective/anti-inflammatory role in the colon under normal and pathological conditions. PMID- 9403786 TI - Growth hormone and somatostatin interaction in the ulcerogenic effect of cysteamine in female rats. AB - It is known that cysteamine's ulcerogenic effect depends, among others, on a depletion of somatostatin (SRIH). Since growth hormone (GH) affects the release of hypothalamic SRIH, we have studied the influence of GH and the GH-SRIH interaction on the severity of gastric mucosa lesions induced by cysteamine. Female rats of the Sprague-Dawley strain were pretreated with GH (1 mg/kg) and subjected to cysteamine-induced gastric lesions. We found that these animals showed an increased mortality and severity of gastric lesions. Pretreatment with SRIH (25 or 50 micrograms/kg) was followed by a decrease in mortality and of incidence and severity of gastric mucosa lesions as compared to those found in control animals pretreated with saline. The dose of 5 micrograms/kg was ineffective in this respect. The combined administration of GH and SRIH revealed that cysteamine ulcerogenic action remained unchanged. It is possible that high levels of plasma GH, as induced by exogenous GH administration, may decrease the release of gastro-intestinal SRIH and this in turn may potentiate the ulcerogenic activity of cysteamine. PMID- 9403787 TI - Role of nitric oxide in pathogenesis of gastric mucosal damage induced by compound 48/80 in rats. AB - We examined the effects of various nitric oxide synthase (NOS) inhibitors on development of gastric lesions induced by compound 48/80 (48/80) in rats and investigated the roles of NO and inducible NOS (iNOS) in inflammatory gastric responses. Animals were given 48/80 (1 mg/kg, i.p.) once daily for 4 days, and the stomachs were examined for lesions 24 h after the final administration. NOS inhibitors such as L-NAME, L-NMMA, aminoguanidine or dexamethasone were administered for 4 days during 48/80 treatment. The repeated administration of 48/80 caused damage in the stomach with severe edema in the submucosa. These lesions induced by 48/80 were dose-dependently prevented by concurrent administration of L-NAME. The protective effect of L-NAME on 48/80-induced gastric lesions was mimicked by L-NMMA, aminoguanidine as well as dexamethasone, and significantly antagonized by co-administration of L-arginine but not by D arginine. Acid secretion was slightly decreased after 48/80 treatment, but was significantly augmented by the combined administration of L-NAME with 48/80. The mucosal MPO activity, TBA reactants and vascular permeability in the stomach were all increased after 48/80 treatment, but these changes were also significantly mitigated by co-administration of L-NAME. The Ca(2+)-independent NOS activity in the mucosa was increased four times during 48/80 treatment, and this change was also inhibited by dexamethasone. These results suggest that: 1) the repeated administration of 48/80 induced inflammatory gastric lesions in the rat stomach; 2) the pathogenic mechanism of these lesions involves endogenous NO produced by iNOS, in addition to oxyradical formation; and 3) the deleterious role of NO during 48/80 treatment may be accounted for by a cytotoxic action of peroxynitrite, which is formed in the presence of NO and superoxide radicals. PMID- 9403788 TI - The influence of BPC 157 on nitric oxide agonist and antagonist induced lesions in broiler chicks. AB - We describe the effects of nitric oxide (NO) agonists and antagonists and the influence of a novel organoprotective pentadecapeptide BPC 157, on the development of pulmonary hypertension syndrome and tissue lesions in chicks. Acute toxicity, which includes single dose application of saline (1 mL intraperitoneally (i.p.)), BPC 157 (10 micrograms/kg bw), L-NAME (NO antagonist, doses 50, 100, 150 mg/kg bw) and L-arginine (NO agonist/100 mg/kg bw with their combination L-NAME + BPC 157; L-NAME + L-arginine) was investigated. In this experiment pathohistological examination of the spleen, heart, liver and lungs and hematological analysis was conducted. In the chronic toxicity experiment, the animals were treated daily for 5 weeks with L-NAME (10 mg/kg bw), L-arginine (100 mg/kg bw), BPC 157 (10 micrograms/kg bw) and their combinations (L-NAME + BPC 157; L-NAME + BPC 157; L-NAME + L-arginine) i.p. Seven animals from each group, including controls (saline 1 mL i.p.) were killed every week. Application of L NAME caused pulmonary hypertension syndrome (PHS) in the treated chicks, which was prevented by the simultaneous application of L-arginine and BPC 157. Pathohistological examination of both acute and chronic toxicity revealed that L NAME caused severe tissue damage (myocardial and hepatic cell necrosis, necrosis of the lymphoid cells in the spleen) while L-arginine provoked predominantly congestion, edema and hemorrhages in all organs. The effect of L-NAME was successfully inhibited by the application of L-arginine and BPC 157 but the latter substance did not cause any tissue or organ damage. Hematological analysis shows significant hemoglobin and leukocyte number decrease in the L-NAME-treated groups of chicks. PMID- 9403789 TI - Capsaicin and the stomach. A review of experimental and clinical data. AB - Capsaicin, the pungent principle of hot pepper, because of its ability to excite and later defunctionalize a subset of primary afferent neurons, has been extensively used as a probe to elucidate the function of these sensory neurons in a number of physiological processes. In the rat stomach, experimental data provided clear evidence that capsaicin-sensitive (CS) sensory nerves are involved in a local defense mechanism against gastric ulcer. Stimulation of CS sensory nerves with low intragastric concentrations of capsaicin protected the rat gastric mucosa against injury produced by different ulcerogenic agents. High local desensitizing concentrations of capsaicin or systemic neurotoxic doses of the agent markedly enhanced the susceptibility of the rat gastric mucosa to later noxious challenge. Resiniferatoxin, a potent analogue of capsaicin possesses an acute gastroprotective effect similar to that of capsaicin in the stomach. The gastroprotective effect of capsaicin-type agents involves an enhancement of the microcirculation effected through the release of mediator peptides from the sensory nerve terminals with calcitonin gene-related peptide being the most likely candidate implicated. They do not depend on vagal efferent or sympathetic neurons or involve prostanoids. The gastric mucosal protective effect of prostacyclin is retained after systemic or topical capsaicin desensitization. Capsaicin-sensitive fibers are involved in the repair mechanisms of the gastric mucosa. A protective role for CS sensory nerves has also been demonstrated in the colon. In most studies, capsaicin given into the stomach of rats or cats inhibited gastric acid secretion. In humans, although recent studies provide evidence in favor of a beneficial effect of capsaicin on the gastric mucosa, an exact concentration-related assessment of the effect of the agent is still lacking. PMID- 9403790 TI - BPC 157's effect on healing. AB - The 15 amino acid agent BPC 157, showing a wide range of organoprotective action in different experimental models, was used in our experiments in order to establish its influence on different elements connected with the healing process. Elements thought to be of greatest importance in the process of healing are formation of granulation tissue, angiogenesis and production of collagen. In our work we tested the influence of BPC 157 on: granulation tissue and collagen formation, on angiogenesis as well as on tensile strength development, using three experimental rat models: 1) skin incisional wounds; 2) colon-colon anastomoses; and 3) angiogenesis model with synthetic sponge implantation. The specimens were histologically assessed for collagen, reticulin and blood vessels using scoring and morphometry. In all experiments significant differences between BPC 157-treated animals and controls were found, showing a strong, promoting involvement of BPC in the healing process. It is worth noting that these effects were achieved by different routes of application, including intragastric and local, making BPC 157 a potentially useful therapeutic agent. PMID- 9403791 TI - Some aspects of the effects of PL-10.1.AK-15 on the gastrointestinal tract. AB - PL-10.1.AK-15 is an active fragment of a naturally occurring protein first isolated from human gastric juice. Among its other protective effects, PL-10.1.AK 15 has demonstrated a protective effect on the gastrointestinal tract. The aim of this study was to investigate the influence of PL-10.1.AK-15 on two functional parameters of gastrointestinal function: gastric acid secretion and gastrointestinal motility. Gastric acid secretion was assessed in male Wistar rats using a modified method of Shay, while gastrointestinal motility was assessed in male NMRI mice by charcoal propulsion. PL-10.1.AK-15 was given in three different doses (3, 10 and 100 micrograms/kg body weight) in accordance with the experimental protocol. The results of these experiments indicate that PL 10.1.AK-15 in the investigated doses had no influence on gastric acid secretion or gastrointestinal motility. PMID- 9403793 TI - A study of the actions of naloxone and morphine on gastric acid secretion and gastric mucosal damage in the rat. AB - There exists a considerable controversy in the literature with regard to the effect of either opiate receptor blockade or that of morphine in different gastric and intestinal ulcer models in the rat. We performed experiments to evaluate the effects of naloxone and morphine on gastric acid secretion and gastric mucosal damage in different experimental models of gastric mucosal injury, namely in indomethacin-, HCl (0.6N)- and ethanol (96%)-models. We found that: 1) 10 mg/kg naloxone i.p. given twice, effectively protected gastric mucosa against indomethacin (30 mg/kg i.p.) and against the acid-dependent injury caused by 0.6 N HCl (1 mL i.g.), but not against the non acid-dependent injury caused by 96% ethanol (1 mL i.g.); 2) morphine (10 + 10 mg/kg i.p.) increased ulcers in the HCl-model, but had no effect in the two other models; 3) this ulcer-aggravating effect of morphine in the HCl-model was blocked by pretreatment of 2 mg/kg i.p. naloxone; and 4) both naloxone (5 + 5 and 10 + 10 mg/kg i.p.) and morphine (10 + 10 mg/kg i.p.) significantly decreased gastric acid secretion in 1-h pylorus ligated rats. We conclude that: 1) naloxone dose-dependently protects against the indomethacin- and HCl-, but not against the ethanol-induced gastric mucosal damage; 2) morphine aggravates the HCl-induced ulcerogenesis; and 3) both opioid receptor agonist and antagonist decrease gastric acid secretion. PMID- 9403792 TI - Direct cellular effects of some mediators, hormones and growth factor-like agents on denervated (isolated) rat gastric mucosal cells. AB - The brain-gut axis has an important role in the mechanism of gastric cytoprotection in vivo. The aim of this study was to evaluate the in vitro effect of protective agents without any central and peripheral innervation. A mixed population of rat gastric mucosal cells was isolated by the method of Nagy et al (Gastroenterology (1994) 77, 433-443). Cells were incubated for 60 min with cytoprotective drugs such as prostacyclin, histamine, pentagastrin and PL-10 substances (synthesized parts of BPC). At the end of this incubation cells were treated by 15% ethanol for 5 min. Cell viability was tested by trypan blue exclusion test and succinic dehydrogenase activity. The following results were obtained: 1) prostacyclin, histamine and pentagastrin had no direct cytoprotective effect on isolated cells; and 2) PL-10 substances significantly protected the cells against ethanol-induced cellular damage. This led to the following conclusions: 1) in the phenomenon of gastric cytoprotection only the growth factor-like agents have a direct cellular effect; and 2) the intact peripheral innervation is basically necessary for the development of mediators and hormone-induced gastric cytoprotection. PMID- 9403794 TI - Different mechanisms are responsible for the contractile effects of histaminergic compounds on isolated intestinal smooth muscle cells. AB - The effects of histamine and dimaprit on intestinal smooth muscle contractility were investigated on isolated cells from longitudinal muscle of the guinea pig ileum. Both histamine (10(-14)-10(-10) M) and dimaprit (10(-13)-10(-10) M) exerted a concentration-dependent contraction of intestinal cells, causing a maximum decrease in cell length of about 20%. This effect was not significantly different from that induced by cholecystokinin-octapeptide (CCK-8) 10(-9) M. The concentration-response curves to histamine and dimaprit were shifted to the left in the presence of the histamine H2-receptor antagonist famotidine (10(-7) M) indicating the occurrence in the smooth muscle of H2 receptors mediating relaxation. Whereas the contraction produced by histamine was competitively antagonized by the H1 receptor antagonist mepyramine (10(-8) M), neither mepyramine (10(-7) M) nor temelastine (10(-7) M) did modify the contractile effect of dimaprit. In contrast, atropine (10(-8) M) significantly depressed the maximum response to dimaprit without affecting that exerted by histamine. These data indicate that histamine and dimaprit can modify intestinal contractility, by acting via different mechanisms; while the contractile action of histamine is related to H1 receptor activation, that produced by dimaprit involves cholinergic pathways. PMID- 9403795 TI - Evidence for a dual mechanism of gastric motor responses to intravenously administered endothelin-1 in anesthetized rats. AB - We have recently reported that endothelin-1 (ET-1), administered intracisternally or microinjected into the DVC of rats, increases gastric motor function via vagal pathways. To determine whether circulating ET-1 acts peripherally or centrally to alter gastric motility, ET-1 (30 and 300 pmol/kg) was administered intravenously in alpha-chloralose anesthetized rats, while monitoring intragastric pressure, gastric motility, heart rate and blood pressure. Endothelin-1, at a dose of 300 pmol/kg, increased intragastric pressure, stimulated pyloric circular muscle contractile activity, and increased arterial pressure. When ET-1 (300 pmol/kg) was administered after bilateral vagotomy at midcervical level, a marked gastric motor inhibition with an increase in arterial blood pressure were observed. We conclude that the gastric motor effects of circulating ET-1 are a result of central excitatory and peripheral inhibitory actions of the peptide. PMID- 9403797 TI - Nitric oxide synthase, oxytocin and vasopressin immunoreactivities in the paraventricular, supraoptic and vagal nuclei of the ferret. PMID- 9403796 TI - Cyclooxygenase inhibition in the dorsal vagal complex of the rat evokes increases in gastric motor function. AB - To characterize the involvement of brainstem cyclooxygenase (COX) in the vagal control of gastric motor function, tolmetin, a reversible COX inhibitor, was applied to the surface of the dorsal medulla oblongata or microinjected into the dorsal vagal complex (DVC) in alpha-chloralose anesthetized rats, while intragastric pressure and contractile activity of the pyloric circular and greater curvature longitudinal muscle were monitored. Tolmetin, applied to the surface of the medulla oblongata, increased intragastric pressure and stimulated contractile activity of gastric smooth muscle. Comparable gastric motor effects were observed after microinjection of tolmetin into the DVC. All the effects of tolmetin were abolished by bilateral vagotomy at the midcervical level. These results demonstrate for the first time that COX inhibition evokes vagally mediated gastric motor effects in the DVC of the rat and support a role for COX products in gastrointestinal regulation. PMID- 9403798 TI - The effect of L-arginine/nitric oxide pathway on salivary amylase secretion in conscious rats. AB - In a recent study we have demonstrated the presence of nitric oxide synthase immunoreactive neurons and also perivascular, periacinar and periductal nerve fibres in feline submandibular salivary gland. The role of nitric oxide (NO) in salivary vasoregulation has been suggested by other data too, but the effect of NO on salivary amylase secretion has not been investigated yet. Under ether anaesthesia a catheter was introduced into the oesophagus for salivary juice collections, and a cannula was inserted into the jugular vein for infusions. After postanaesthesia recovery, submaximal carbachol infusion was given as a background to obtain steady secretion because of the low basal secretory rate. Then different groups of animals received NO synthase inhibitor NOLA (NG-nitro-L arginine), L-arginine, D-arginine or NO donor SIN-1 (3-morpholinosydnonimine). Volume and amylase activity were determined in mixed saliva samples collected for 30 min. Carbachol background infusion alone induced an elevated, sustained salivary secretion. NOLA (30 mg/kg) increased both volume and amylase output (P < 0.001). This effect was prevented by L-arginine but not by D-arginine. SIN-1 did not change either volume or amylase secretion. The present results suggest that the L-arginine/NO pathway has a modulatory effect on salivary fluid and amylase secretion, which is probably not related to its effect on salivary blood flow. NO may block certain presently unidentified secretagogue mechanisms and/or may relax myoepithelial cells. PMID- 9403799 TI - Effect of somatostatin immunoneutralization on gastric acid and pancreatic enzyme secretion in anesthetized rats. AB - The involvement of somatostatin in urethane-anesthesia-evoked suppression of gastric acid secretion has been described. The present study has examined the role of endogenous somatostatin in diminished pancreatic enzyme secretion during anesthesia, while monitoring acid secretion concurrently. Rats were anesthetized with either urethane or sodium pentobarbital. An indwelling catheter was placed into the right jugular vein. The esophagus and the pylorus were ligated, and the stomach was perfused with saline. The common bile duct was ligated at the hepatic hilum, and cannulated at the duodenal end of the duct for collecting pure pancreatic juice. Purified somatostatin monoclonal antibody (CURE.S6) or control antibody (keyhole limpet hemacyanin, KLH) was injected iv in increasing doses (0.05; 0.15; 0.5; and 1.5 mg) every 30 min (n = 6). Gastric acid and pancreatic amylase secretions were measured. The effect of the antibodies on CCK-8 stimulated (0.25-2.50 nmol/kg/h) pancreatic amylase secretion was also tested. During urethane anesthesia somatostatin antibody induced a dose-dependent increase in acid output, while control antibody did not change it. Basal pancreatic amylase secretion was not affected by either somatostatin or by control antibody. Pancreatic secretory responses to high but not to low doses CCK 8 were found to be significantly increased following immunoneutralization of somatostatin. In sodium pentobarbital-anesthetized rats somatostatin antibody stimulated basal acid secretion but did not affect basal pancreatic amylase secretion. Our data indicate that in anesthetized rats endogenous somatostatin mediates suppression of basal gastric acid secretion but not that of basal pancreatic amylase secretion, and this action does not depend on the type of anesthesia. Furthermore, endogenous somatostatin may play a physiological role in modulating stimulated pancreatic enzyme secretion in this species. PMID- 9403800 TI - Stimulatory effect of PACAP on gastroduodenal bicarbonate secretion in rats. AB - The effects of pituitary adenylate cyclase activating polypeptides (PACAPs) on gastroduodenal HCO3- secretion were investigated in anesthetized rats and compared with those of vasoactive intestinal polypeptide (VIP). Under urethane anesthesia, a rat stomach mounted in an ex vivo chamber (in the absence of acid secretion) or a rat proximal duodenal loop was perfused with saline, and the HCO3 secretion was measured at pH 7.0 using a pH-stat method and by adding 10 mM HCl. Intravenous injection of PACAP-27 stimulated HCO3- secretion in a dose-dependent manner in the duodenum but not in the stomach; at 8 nmol/kg PACAP-27 increased the HCO3- secretion to maximal values of four times greater than basal levels, although this peptide had no effect on duodenal HCO3- secretion after intracisternal administration (1 nmol/rat). PGE2 (300 micrograms/kg, iv) significantly increased HCO3- secretion in both the stomach and the duodenum. The potency of duodenal HCO3- secretory action was in the following order; PACAP-27 > PACAP-38 = VIP, and that of PACAP-27 was about 100-fold greater than that of PGE2. The duodenal HCO3- secretory action of PACAP-27 as well as PGE2 was markedly potentiated by prior administration of isobutylmethyl xanthine (10 mg/kg, sc), the inhibitor of phosphodiesterase. Folskolin (250 micrograms/kg, iv), the stimulator of adenylate cyclase, also increased HCO3- secretion in the duodenum but not in the stomach. These results suggest that: 1) PACAPs are potent stimulators of HCO3- secretion in the duodenum but not in the stomach; 2) this action is mediated by cAMP through stimulation of adenylate cyclase; 3) cAMP is a mediator in duodenal but not gastric HCO3- secretion; and 4) PACAPs may be involved in the peripheral regulation of duodenal HCO3- secretion. PMID- 9403801 TI - Impaired duodenal bicarbonate secretion in diabetic rats. Salutary effect of nitric oxide synthase inhibitor. AB - We previously reported the impaired HCO3- secretion and the increased mucosal susceptibility to acid in the duodenum of streptozotocin (STZ)-induced diabetic rats. In this study, we investigated the salutary effect of the NO synthase inhibitor L-NAME (NG-nitro-L-arginine methyl ester) on these changes and compared it with those of insulin. Animals were injected streptozotocin (STZ: 70 mg/kg, ip) and used after 1, 3-4, and 5-6 weeks of diabetes with blood glucose levels of > 300 mg/dL. Under urethane anesthesia the HCO3- secretion was measured in the proximal duodenal loop using a pH-stat method and by adding 10 mM HCl. L-NAME (20 mg/kg x 2) or insulin (4 units/rat) was administered sc for 4-5 weeks, starting 1 week after STZ treatment. The duodenal HCO3- secretory responses to various stimuli such as mucosal acidification (10 mM HCl for 10 min), 16,16-dimethyl prostaglandin E2 (dmPGE2: 10 micrograms/kg, i.v.), and vagal stimulation (0.5 mA, 2 ms, 3 Hz) were significantly decreased in STZ-treated rats, depending on the duration of diabetes. Repeated administration of L-NAME, starting from 1 week after STZ treatment, significantly reduced blood glucose levels toward normal values and restored the HCO3- responses to various stimuli in STZ rats, the effects being similar to those observed after supplementation of insulin. Diabetic rats developed duodenal lesions after perfusion of the duodenum with 150 mM HCl for 4 h, but this ulcerogenic response was significantly inhibited by the repeated treatment with L-NAME as well as insulin. We conclude that L-NAME is effective in ameliorating hyperglycemic conditions in STZ-diabetic rats, similar to insulin, and restores the impaired HCO3- secretion and the increased mucosal susceptibility to acid in diabetic rat duodenums. PMID- 9403802 TI - Comparison between beta 3 and beta 2 adrenoceptor agonists as inhibitors of gastric acid secretion. AB - In order to investigate the role of beta 3 adrenoceptors in the regulation of gastric acid secretion we studied the effects of compound SR58611A (a selective agonist for atypical beta adrenoceptors), alone or in combination with beta adrenoceptor antagonists, in the gastric fistula of a conscious cat. The effects of SR58611A were compared with those of clenbuterol, a selective agonist for beta 2 adrenoceptors. Intravenous infusion of SR58611A (0.3-3 mumol/kg/h) caused a dose-dependent, but partial, inhibition of the acid secretory response to 2-deoxy D-glucose 100 mg/kg i.v., maximum effect not exceeding 40%. Clenbuterol (0.03-0.1 mumol/kg/h) caused a similar effect (maximum inhibition about 50%) at doses approximately 30 times lower. The acid secretion induced by the histamine H2 receptor agonist dimaprit (1 mumol/kg/h) was minimally affected by both beta adrenoceptor agonists. The inhibitory effect of SR58611A (3 mumol/kg/h) on 2 deoxy-D-glucose-induced acid secretion was not modified by pretreatment with the non-selective beta 1- and beta 2-adrenoceptor blocker propranolol, administered at doses (1.5 mumol/kg iv) that completely blocked the inhibitory effect of clenbuterol (0.1 mumol/kg/h). In contrast, bupranolol (10 mumol/kg i.v.) (a drug endowed with beta 3 antagonistic properties) prevented the inhibitory effects of both SR58611A and clenbuterol. The present data provide functional evidence that, besides beta 2-, also beta 3-adrenoceptors can have negative effects on gastric acid secretion, particularly when it is stimulated by indirect stimuli, like 2 deoxy-D-glucose. This gastric antisecretory activity may represent an additional mechanism for the physio-pharmacological control of gastric acid secretion. PMID- 9403803 TI - Dopaminergic characteristics of isolated parietal cells from rats. AB - Recently we have identified a dopamine-producing system in the gastric mucosa of rats. All the available morphological data suggest that parietal cells synthesize dopamine. In the present study we investigated the dopaminergic characteristics of isolated parietal cells by different methods. Mixed gastric mucosal cells were isolated and size-fractionated by elutriation. The proportion of neurons, parietal and endocrine cells in the fractions were determined by immunocytochemistry (ICC) using antibodies to neurofilament, proton pump and chromogranin A, respectively. No neurons were found in any of the cell preparations, while 56% parietal cell and 0.0% endocrine cell were achieved in the parietally enriched fraction. By Western blot, a tyrosine hydroxylase (TH, the rate-limiting enzyme of the catecholamine synthesis) immunoreactive protein species was demonstrated in isolated mucosal cells, comigrating with the TH immunoreactivity from PC12 cells. The TH immunoreactivity was colocalized to parietal cells by ICC. Dopamine transporter (DAT), a regulator of extracellular/intracellular dopamine balance in the nervous system, was also demonstrated in parietal cells. A significant amount of dopamine and DOPA were measured by HPLC (13.4 and 9.57 pg/10(6) cell, respectively) in parietally enriched cell fraction. Since this enriched cell fraction was virtually clear of both neurons and endocrine cells, demonstration of TH enzyme, DAT and dopamine in this fraction confirms that the parietal cell population might be a major source of dopamine in the rat stomach, supporting our previous results achieved using whole tissue samples. PMID- 9403804 TI - Effect of a new CCK-A receptor antagonist, dexloxiglumide, on the exocrine pancreas in the rat. AB - The effect of dexloxiglumide, a new potent cholecystokinin (CCK) antagonist, on pancreatic enzyme secretion and growth was studied in the rat. Pancreatic exocrine secretion was studied both in vitro (isolated and perfused pancreatic segments) and in vivo (anaesthetized animals with cannulation of the common bile duct) whereas the trophic effect was investigated after short-term (7 days) administration of the CCK-agonist, caerulein, or camostate (a potent trypsin inhibitor), with or without dexloxiglumide. CCK-8 stimulated amylase release from in vitro pancreatic segments in a concentration-dependent manner. Dexloxiglumide displaced the concentration response curves to CCK-8 to the right without affecting the maximum response, suggesting a competitive antagonism. The Schild plot analysis of data gave a straight line with a slope (0.90 +/- 0.36) not significantly different from unity. The calculated pA2 for dexloxiglumide was 6.41 +/- 0.38. In vivo experiments confirmed results from in vitro studies since intravenous dexloxiglumide reduced pancreatic exocrine secretion induced by submaximal CCK-8 stimulation (0.5 nmol/kg/h) in a dose-dependent manner, the ID50 being 0.64 mg/kg. Both exogenous and endogenous (released by camostate) CCK increased the weight of the pancreas, the total pancreatic protein and DNA, trypsin and amylase content. Dexloxiglumide (25 mg/kg), administered together with caerulein (1 microgram/kg), reduced the peptide-induced increase in pancreatic weight, protein and enzyme content. Similarly, when dexloxiglumide was given together with camostate (200 mg/kg), all the observed changes were reduced by concomitant administration of the antagonist. These results demonstrate the ability of dexloxiglumide to antagonize the effects of CCK on pancreatic secretion and growth, suggesting that this compound is a potent and selective antagonist of CCK-A-receptors in the pancreas. PMID- 9403805 TI - Effects of TRH on gastric acid secretion: a model for human study. AB - The possible effects of TRH administration on different parameters of gastric function were studied in 10 patients with different gastrointestinal complaints. Basal (BAO) and pentagastrin stimulated (6 micrograms pentagastrin/kg bw sc) maximal (MAO) acid output were determined and serum levels of TSH, total and free thyroxine (T4 and FT4), triiodothyronine (T3) were measured. After determinations of BAO and MAO and the hormones indicated above, one group of patients received a TRH injection (0.2 mg protirelin) intravenously. The second group of patients was injected with atropine (atropinum sulfuricum, 1 mg, iv). At different times following the injections in both groups of patients BAO, MAO and serum levels of TSH, total and free T4, T3, gastrin were determined. Injection of TRH resulted in an increase in TSH and with some delay in thyroxine and gastric acid levels. Atropine treatment was followed by a decrease in gastric acid secretion and a small decrease in TSH and no changes in the values of the other studied hormones. The results suggest a complex interrelationship between TRH, vagal system and pentagastrin-dependent gastric acid secretion operating in human subjects. PMID- 9403806 TI - What is your diagnosis? Erythroderma form of mycosis fungoides. PMID- 9403808 TI - Incidence of contralateral versus ipsilateral neurological signs associated with lateralised Hansen type I disc extrusion. AB - Asymmetrical neurological signs were noted in 50 dogs presenting with Hansen type I thoracolumbar disc extrusion. Thoracolumbar myelograms and surgical decompression were performed in all cases. Dogs were divided into two groups (acute and chronic) based on the duration of clinical signs prior to presentation to the University of Georgia. Lateralising extradural cord compressive lesions were noted on all myelograms. In the acute group, 35 per cent of the dogs had asymmetrical neurological signs contralateral to the myelographic and surgical lesion, while in the chronic group only 11 per cent had neurological signs contralateral to the lesion. There was found to be no significant difference in frequency of contralateral asymmetrical clinical signs between the two groups (Fischer's exact test; P = 0.095). The high frequency of contralateral signs documents the importance of thoracolumbar myelography for accurate localisation of the disc material before decompressive surgery. PMID- 9403807 TI - Hemilaminectomy for the treatment of thoracolumbar disc disease in the dog: a follow-up study of 40 cases. AB - A study was made of dogs with Hansen type I thoracolumbar disc extrusions that had been treated by hemilaminectomy and fenestration of the affected disc. Follow up information was available for 40 dogs undergoing treatment over a five-year period. The follow-up period ranged from 12 to 72 months (mean 34 months). The case details and the results of treatment of these 40 dogs are presented. All dogs were graded according to the degree of neurological dysfunction at the time of initial presentation and at the conclusion of the study period. Twenty-seven dogs (68 per cent) had no detectable signs of neurological dysfunction or thoracolumbar pain at the final assessment and a further eight dogs (20 per cent) had mild ambulatory paraparesis but were regarded by their owners as functional pets. Recurrence of neurological signs consistent with thoracolumbar disc disease was seen in five dogs (13 per cent) and was successfully resolved completely in one of three dogs that were treated. PMID- 9403809 TI - Hereditary multifocal renal cystadenocarcinomas and nodular dermatofibrosis in 51 German shepherd dogs. AB - The clinical findings in German shepherd dogs with hereditary multifocal renal cystadenocarcinomas and nodular dermatofibrosis are presented. Between 1978 and 1996, 51 cases were examined. Eight cases were detected after being offered a clinical examination because the disease was present in a parent. The remaining 43 dogs were diagnosed after an unsolicited visit to a clinic because of a specific problem. Skin lesions were the main reason (37 per cent) the owners presented their dog for examination. The mean age at diagnosis of renal cystadenocarcinomas and nodular dermatofibrosis was 8.2 years. The male-to-female ratio was 1.1, while the corresponding figure for a reference population was 1.25. Enlarged and abnormally shaped kidneys were palpated in 60 per cent of the dogs and were detected by radiography in 86 per cent of cases. The renal lesions, including metastases, were the main reason for euthanasia and death. The mean age at death was 9.3 years, and the mean age at the first detection of nodular dermatofibrosis was 6.4 years. PMID- 9403810 TI - Testicular fine needle aspiration cytology as a diagnostic tool in dog infertility. AB - The results of testicular aspirate cytology taken from clinical patients with a history of infertility were compared with the clinical and histological findings. Azoospermia was the most common and the most rewarding indication for the examination. Samples were also taken from cases with suspected testicular tumours, orchitis, epididymitis, severe oligo- and teratozoospermia, lack of libido and unilateral testicular atrophy. Histological and cytological findings were found to correlate well. Identification of cell types from normal germinal epithelium was relatively easy. No immediate adverse effects of aspiration were noted. Five normospermic dogs were monitored for two to six months after aspiration, and there were no marked deleterious effects on testicular consistency, testicular histology or semen characteristics. Testicular cytology obtained by fine needle aspiration may, at least to some extent, be used to assist clinical diagnosis, especially in azoospermic dogs and dogs with palpable changes of testicular tissue. PMID- 9403811 TI - Use of ultrasonography in the diagnosis of tracheal collapse. AB - Ultrasonographic imaging of the cervical trachea was performed with the neck in both a neutral and a hyperextended position in 10 dogs with tracheal collapse. Tracheoscopy was used to confirm a diagnosis of tracheal collapse. The ultrasound investigation was repeated in 10 dogs of similar size but without tracheal abnormality. The ultrasonographic findings of the affected dogs were compared with those of the normal group and showed an alteration in the shape of the tracheal lumen in the ventrodorsal projection. This study highlights the possibility of identifying changes in the shape of the tracheal lumen during ultrasound investigations as an aid to the diagnosis of tracheal collapse. PMID- 9403812 TI - Mitotane (o,p'-DDD) treatment in a cat with hyperadrenocorticism. AB - An 11-year-old male castrated Persian cat with spontaneous hyperadrenocorticism was presented. Both adrenals were grossly enlarged and calcified. A diagnosis of pituitary-dependent hyperadrenocorticism was made. Signs of hyperadrenocorticism resolved with long-term mitotane treatment. Concurrent diabetes mellitus resolved after 220 days of therapy. No severe adverse drug reactions were noted. PMID- 9403813 TI - Persistent left cranial vena cava associated with multiple congenital anomalies in a six-week-old puppy. AB - A six-week-old male puppy was presented with a distended abdomen, dypsnoea and cyanosis. Auscultation revealed a grade II/VI systolic murmur. Thoracic radiographs showed gross cardiomegaly. An electrocardiogram revealed a narrow complex tachycardia, deep S waves in leads I, II, III and aVF, and negative P waves in lead III. Two-dimensional echocardiography showed a high ventricular septal defect and marked dilation of the right-sided chambers. There was also an echolucent structure lateral to the left atrium at a site corresponding to the coronary sinus. Contrast echocardiography revealed right-to-left shunting through the septal defect. Necropsy confirmed the existence of a septal defect in the membranous part of the septum and a persistent left cranial vena cava with dilation of the coronary sinus. In addition, a small patent ductus arteriosus and tricuspid dysplasia were present. PMID- 9403814 TI - Toxocara canis. PMID- 9403815 TI - Gastrointestinal hormone in dumping syndrome and reflux esophagitis after gastric surgery. AB - Several problems are associated with gastric resection, including the dumping syndrome, reflux esophagitis, and malabsorption. A better understanding of the pathophysiological changes will shed light on new and improved therapy. Serum levels of seven circulating gastrointestinal hormones following a standardized solid meal and a brief score of symptoms were evaluated in 10 patients after partial distal gastrectomy and 12 patients after total gastrectomy, both groups reconstructed by Billroth II anastomosis, and 9 age-matched healthy controls. Patients underwent resection for gastric cancer and were studied 45 +/- 10 months after surgery. At the time of study, the patients had adapted well to surgery and no longer exhibited the severe symptoms of dumping seen immediately post operatively. In contrast, the total gastrectomy patients exhibited the symptoms of reflux esophagitis. The gastrointestinal hormone changes could be divided into three patterns; obtunded responses (gastrin, PP), normal release (motilin, GIP) and increased secretion (CCK, neurotensin, PYY). In these, the early reaction of neurotensin correlated with the scores of late dumping syndrome and reflux esophagitis. In the literature, many gastrointestinal hormones have been shown to respond as an enhancement rather than adaptation. In other gastrointestinal hormones, secretin belonged to the obtunded type and enteroglucagon were classified in the increased type. However, pathophysiological significance of these hormonal changes remained uncertain. The late adaptive changes in gastrointestinal hormone secretion may help to compensate for loss of gastric motor function which accompanies gastric resection. On the other hand, these hormonal changes may exacerbate the esophageal reflux following gastrectomy. PMID- 9403816 TI - Inhibitory actions of endothelial products on mechanical and calcium responses in aortic smooth muscle cells. AB - The effects of endothelial products on intracellular calcium ion concentrations ([Ca2+]i) were investigated in the guinea-pig aorta. The perfusate of bradykinin stimulated cultured endothelial cells relaxed aortic rings by approximately 70%; it was reduced to approximately 50% by nitroarginine, to approximately 30% in high-K solution and remained unaltered by indomethacin. The perfusate elevated the cyclic GMP content in the muscle, which was inhibited by nitroarginine. In cultured, aortic muscle cells, bradykinin elevated [Ca2+]i with an initial transient and following sustained phase; the former was absent after treatment with cyclopiazonic acid while the latter was abolished in [Ca2+]o-free medium. The perfusate lowered aortic [Ca2+]i, and this action was weakened by nitroarginine and diminished in high-K solution. Therefore, the perfusate reduced aortic [Ca2+]i with and without an increase in cyclic GMP production. These actions were sensitive to nitroarginine and high-K, respectively, suggesting that the perfusate contained at least two relaxants, EDRF and EDHF, with both lowering [Ca2+]i in aortic muscle mainly by inhibiting Ca2+ influx. PMID- 9403817 TI - HPC-1/syntaxin-1A activity in the enteric nervous system of developing rat gastrointestinal tract. AB - The HPC-1/syntaxin-1A antigen was originally identified as a neuron-specific membrane protein in the central nervous system. The presence of HPC-1 antigen in the nervous system of the fetal rat gastrointestinal tract was immunohistochemically demonstrated using the antibody against HPC-1 to clarify the role of this protein in the development of the enteric nervous system. Rat gastrointestinal tract from 14-, 16-, 18-, and 20-day fetuses and adults were immunohistochemically examined for HPC-1 antigen by light microscopy. Acetylcholinesterase (AchE) activity was also examined as a comparison. HPC-1 activity was first detected on 18th day of gestation. AchE activity was first detected at the Auerbach's plexus of the esophagus on the 16th day of gestation. The presence of HPC-1 in the developing rat intestine revealed that the HPC-1 antigen may be a good indicator for expressing the maturation of enteric nervous system in the development of the enteric nervous system. PMID- 9403818 TI - Comparison of the relaxing actions of acetylcholine and substance P in smooth muscle of the guinea-pig aorta. AB - The relationship between relaxation produced by acetylcholine (ACh) or substance P (SP) and tissue cyclic GMP content was investigated in the isolated guinea-pig aorta. ACh and SP relaxed aortic rings precontracted with noradrenaline (NA) or high-K solution ([K+]o = 38.8 mM), in an endothelium-dependent manner. The amplitude of relaxation was larger for SP than for ACh. Nitroarginine inhibited ACh-induced but not SP-induced relaxation in NA-contraction, while this chemical inhibited both ACh- and SP-induced relaxations in high-K contraction. The tissue cyclic GMP content was not changed by nitroarginine or by removal of endothelial cells, but was elevated by stimulation with NA, ACh or SP by a factor of about 3, 5 or 11 times, respectively. These actions of ACh or SP were endothelium dependent, and were inhibited by nitroarginine and remained unaltered by high-K solution. Thus, ACh and SP relax muscles indirectly by releasing endothelial factors, and the former by releasing mainly an endothelium-derived relaxing factor (EDRF), and the latter by releasing EDRF and other unidentified factors. As the relaxing actions of the latter factors are inhibited by high-K solution with no relation to the production of cyclic GMP, an involvement of hyperpolarizing factor, possibly EDHF, is suggested. PMID- 9403819 TI - Humanitarian aid operations in Republica Srpska during Operation Resolute 2. AB - The humanitarian aid experience of a unit in Bosnia is described. Data are presented for primary care clinics undertaken, showing the range of conditions and age of patients seen. The role of the civilian aid agencies involved is described, together with recommendations for future training requirements for similar operations. PMID- 9403820 TI - The Pre-Hospital Emergency Care Course (PHEC). A TA initiative. AB - The Pre-Hospital Emergency Care (PHEC) course is a 3 day course run by BASICS Education for the Royal College of Surgeons of Edinburgh, Royal College of Nursing and the NHS Ambulance Service. PHEC teaches and examines the care of medical, trauma, paediatric and obstetric patients in the pre-hospital environment. 243 (The Wessex) Field Hospital has held two of an intended ongoing programme of PHEC courses and they have proved suitable for medical and dental officers, nurses and combat medical technicians. We believe the courses to have a valuable role in the ongoing training of CMT is as well as medical and nursing officers. Furthermore, being nationally recognised qualifications we hope the courses will have a positive effect not only on personnel retainment but on recruitment. Demand for places has far exceeded availability and we believe will continue to do so as these courses are not only highly relevant to TA service but to civilian career progression. Furthermore, as the humanitarian role of the armed forces becomes more and more important the PHEC course will become increasingly relevant to regular army personnel not only during their service but also after leaving the forces. The ultimate beneficiaries will be our patients. PMID- 9403821 TI - An evaluation of infra-red tympanic thermometry for thermal physiology research. AB - This study forms part of a research programme to investigate the relationships between the rise in core temperature and the environmental temperature in soldiers exercising at a constant rate. The measurement of core temperature is a fundamental requirement for this research. The aim of this study was to evaluate the use of an infra-red tympanic thermometer (IVAC Corecheck) for use in thermal physiological studies conducted outside the laboratory environment by comparing the right ear temperature by TM thermometer and the left ear external auditory meatus by thermistor. The slope of the regression line between the two measurements was 0.9967 with a correlation coefficient of 0.706 which is reasonable. The TM thermometer in general reads higher than the aural thermometer with a 95% confidence limit for agreement from +0.78 to -0.53 degrees C of the aural temperature. Thus this study demonstrated that the tympanic thermometer (IVAC Corecheck) is sufficiently reliable compared to the aural thermistor to justify field trials. PMID- 9403822 TI - Establishing a new ENT-head and neck clinic. AB - We report our experience of the first fourteen months of a new head and neck surgical clinic established at a district general hospital. The structure of the clinic together with analysis of the patient case mix and referral patterns are considered. The perceived benefits include rapid patient access to specialist management, the development of subspecialty skills and excellent training opportunities for trainee registrars. PMID- 9403823 TI - The chronic prostatitis syndromes. AB - Three chronic prostatitis syndromes are recognised, chronic bacterial prostatitis (CBP), chronic nonbacterial prostatitis (CNBP), and prostatodynia. All may occur in men of military age, and may tax the patience of medical officers and patients whose capacity for full duty will be impaired. Diagnosis depends on identifying micro-organisms in CBP and white cells in CNBP in prostatic secretion (EPS) expressed by prostatic massage. In prostatodynia there are clinical features of prostatitis but no evidence of inflammation. Prostatic massage should be preceded by trans-rectal ultrasound which may show prostatitis and other pathology, and has simplified the investigation of these syndromes. Management includes a high fluid intake, regular bowels with a soft stool, regular prostatic drainage by ejaculation and limited alcohol intake. Antimicrobials are indicated for CBP and probably for CNBP, and need to be continued for at least three months in many cases. Other measures for treating CNBP are less well established. Prostatodynia is an ill defined syndrome which requires careful evaluation and patients may need psychiatric therapy. PMID- 9403824 TI - BATLS course--9 January 1997. PMID- 9403825 TI - Aviation medicine--international conference in the "Windy City". PMID- 9403826 TI - Caribbean itch: eight cases and one who didn't (Exercise Blue Calypso Diamond). AB - Stinging plankton are a potential source of irritation and morbidity for those pursuing aquatic sports in areas where they are numerous. The problem is seasonal, and likely to be familiar to local people. We describe an incident in which a number of military personnel were affected by the stings of the thimble jelly fish (Linuche unguiculata), with a brief review of the biology of the problem and some recommendations for prevention. PMID- 9403827 TI - Acute compartment syndrome--presenting as severe pain in an extremity out of proportion with the injury. AB - A 24 year old combat medic was admitted to the field hospital at Tomislavgrad in Bosnia, with a suspected forearm, fracture. Radiographs did not show any bony injury. Clinical examination showed marked swelling and tenderness over the extensor compartment. The pain became more severe over the following 12 hours with the pain becoming most intensely felt in the extensors on passive extension. Fasciotomy for suspected acute compartment syndrome was carried out. Acute compartment syndrome is a common complication of extremity injury, and is a clinical diagnosis which should be suspected in all injuries with marked swelling and severe pain. PMID- 9403828 TI - Clostridium difficile toxic megacolon following splenectomy. AB - A case of toxic megacolon following splenectomy for lymphoma is presented. The aetiology of Clostridial difficile infection is reviewed and the hazards of perioperative prohylactic antibiotics are discussed. PMID- 9403829 TI - Enterolith ileus as a complication of small bowel diverticulosis. AB - We report a case of small bowel obstruction which was found to be due to an enterolith obstructing the distal ileum in the manner of "gallstone ileus." The enterolith had formed in an acquired jejunal diverticulum. We review the accepted mechanism of formation of these "stones" and propose a protocol for treatment of this rare complication. PMID- 9403830 TI - Early 19th century Maltese doctors in the service of the Crown. AB - The establishment of a British garrison in Malta in September 1800, provided opportunities for Maltese doctors to join the Army Medical Department. This article follows the academic and military life of four Maltese practitioners, whose joint careers span the era of the Napoleonic Wars. PMID- 9403831 TI - Physiology, idiosyncrasy and heat stress limits. PMID- 9403832 TI - 'Total Trauma Care'--trauma care (UK) 1st biennial conference Organising Committee of Trauma Care (UK) PMID- 9403833 TI - Trauma management on the battlefield: a modern approach. PMID- 9403834 TI - Leishmania major infection in C57BL/10 mice differing at the Lps locus: a new non healing phenotype. AB - The course of cutaneous leishmaniasis was examined in mice from two genetically closely related strains, C57BL/10ScCr (Cr) and C57BL/10ScSn (Sn). Sn mice are able to heal Leishmania major infections, while Cr mice are unable to heal. The cutaneous lesions of the Cr mice progressed continuously and the increase in lesion size was paralleled by an unrestricted growth of the parasites in vivo. Cr mice, in contrast to their Sn counterparts, are highly resistant to all effects of lipopolysaccharide (LPS). The nonhealing L. major infection in Cr mice is in sharp contrast to the course of infection in another endotoxin-nonresponder mouse strain, C3H/HeJ, which heal infections with L. major. Cr mice exhibit, in addition to the defective LPS responsiveness, an impaired interferon-gamma (IFN gamma) response after infection with a variety of microorganisms. The insufficient activation of parasitized macrophages to kill intracellular L. major could be due to the inability of splenocytes from infected Cr mice to secrete IFN gamma upon restimulation with L. major. IFN-gamma is essential for the efficient activation of parasitized macrophages to kill intracellular L. major by producing nitric oxide (NO). Although bone marrow-derived Cr macrophage do not produce NO in response to LPS, both Sn and Cr macrophages release NO upon stimulation with IFN-gamma and tumor necrosis factor, indicating that they are responsive to activation by these cytokines. PMID- 9403836 TI - Direct sputum analysis of Pseudomonas aeruginosa macrorestriction fragment genotypes in patients with cystic fibrosis. AB - The distribution of bacterial populations in the airways of 13 patients with cystic fibrosis who were colonized for 6-23 years with Pseudomonas aeruginosa was investigated by genotyping of bacterial chromosomes directly isolated from 21 sputa. After removal of host material from sputum by hypotonic cell lysis and repetitive washing and centrifugation steps, agarose-embedded bacterial cells were lysed, residual eukaryotic DNA separated by field inversion gel electrophoresis, and the purified bacterial chromosomes subjected to macrorestriction fragment pattern and Southern analyses. Bacterial populations consisted of a single P. aeruginosa clone in 17 sputa, of which more than one clonal variant was apparent in two SpeI fragment fingerprints. Two clones of P. aeruginosa and another species co-existed in four samples. Genomically homogeneous populations of P. aeruginosa are characteristic for chronically colonized lungs in most cases of cystic fibrosis. PMID- 9403835 TI - Antirotaviral activity of milk proteins: lactoferrin prevents rotavirus infection in the enterocyte-like cell line HT-29. AB - Different milk proteins were analyzed for their inhibitory effect on either rotavirus-mediated agglutination of human erythrocytes or rotavirus infection of the human enterocyte-like cell line HT-29. Proteins investigated were alpha lactalbumin, beta-lactoglobulin, apo-lactoferrin, and Fe(3+)-lactoferrin, and their antiviral action was compared with the activity of mucin, a milk glycoprotein known to affect rotavirus infection. Results obtained demonstrated that beta-lactoglobulin, apo- and Fe(3+)-lactoferrin are able to inhibit the replication of rotavirus in a dose-dependent manner, apo-lactoferrin being the most active. It was shown that apo-lactoferrin hinders virus attachment to cell receptors since it is able to bind the viral particles and to prevent both rotavirus haemagglutination and viral binding to susceptible cells. Moreover, this protein markedly inhibited rotavirus antigen synthesis and yield in HT-29 cells when added during the viral adsorption step or when it was present in the first hours of infection, suggesting that this protein interferes with the early phases of rotavirus infection. PMID- 9403838 TI - Autoantibodies with a protective function: polyreactive antibodies against alkaline phosphatase in bacterial infections. AB - In patients with acute bacterial infections antibodies directed against a particular bacterial antigen were detected. The molecular mass of this bacterial antigen was 50 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. By comparison of the NH2-terminal amino acid sequence, the 50 kDa antigen was identified as alkaline phosphatase (AP). Affinity-purified antibodies from patient's sera directed against the bacterial AP (anti-alpha) were also shown to react with human and animal AP, which have different structures. Anti-alpha are IgG subtype 3 immunoglobulins, and their light chains are of the kappa type. Upon isoelectric focussing, the anti-alpha formed a scalariform pattern with five to seven bands in the pH range 7-9. The anti-alpha have an opsonic activity and cause a five- to eightfold increase of phagocytosis of gram-positive and gram-negative bacteria. According to their polyreactivity, their sudden rise early in infection, their oligoclonality, as well as their opsonizing properties, they are assumed to be permanently available natural antibodies that take part in early defence mechanisms. PMID- 9403837 TI - Enhancement of human parainfluenza virus-induced cell fusion by pradimicin, a low molecular weight mannose-binding antibiotic. AB - Oligosaccharides, especially mannose residues, expressed on the cell surface, are thought to be important for virus-induced membrane fusion. We examined the effect of mannose-binding compounds, pradimicin derivative BMY-28,864 (PRM) and concanavalin A (Con A), on cell fusion of human parainfluenza type 2 virus (hPIV2)-infected HeLa cells. Syncytium formation of hPIV2-infected HeLa cells was suppressed in the presence of Con A. On the other hand, PRM enhanced cell fusion of hPIV2-infected HeLa cells. These effects were blocked by addition of mannose rich mannan. However, PRM shows little effect on virus growth and the expression of viral glycoproteins on the cell surface in hPIV2-infected HeLa cells. Fluorescein-isothiocyanate-labeled pradimicin and Con A bound to both uninfected and hPIV2-infected mononuclear cells, indicating that these compounds have an affinity to several cellular component(s). In contrast to Con A, PRM had little affinity to the viral glycoproteins. It is inferred from these results that the enhancement of hPIV2-induced cell fusion is probably due to the interaction between PRM and cellular component(s). PMID- 9403839 TI - Protein tyrosine kinase activation provides an early and obligatory signal in anti-FRP-1/CD98/4F2 monoclonal antibody induced cell fusion mediated by HIV gp160. AB - The mechanism by which anti-fusion regulatory protein-1 (FRP-1) monoclonal antibody (mAb) induced cell fusion was investigated using U2ME-7 cells that are CD4+U937 cells transfected with the HIV gp160 gene. Protein kinase inhibitors (H 7, H-89, herbimycin A and genistein) suppressed cell fusion of Cd+U2ME-7 cells induced by anti-FRP-1 mAb. H-7 and H-89 also inhibited the cell aggregation, but herbimycin A and genistein did not. Intriguingly, only when herbimycin A was added either before or simultaneously with addition of anti-FRP-1 mAb, was cell fusion suppressed, suggesting that tyrosine kinase is related with the initial step of polykaryocyte formation. Anti-FRP-1 mAb induced the rapid tyrosine phosphorylation of multiple cellular proteins. These effects occurred within 1 min and returned to near baseline by 60 min. The rapid tyrosine phosphorylation was suppressed by herbimycin A and genistein. Although it remains to be determined which protein tyrosine kinase(s) is involved in this response, pp130 tyrosine phosphorylation appears to be a specific and early signal transmitted after the interaction of FRP-1 with a specific antibody. pp130 was present in the cytosol fraction and was distinct from pp125FAK, p130CAS, vinculin, and beta 1 integrin. Thus, our study may present evidence for a novel pathway of protein tyrosine kinases that phosphorylate specific, still unknown protein substrates during polykaryocyte formation. PMID- 9403840 TI - Cloning and functional characterization of the genes encoding 3-dehydroquinate synthase (aroB) and tRNA-guanine transglycosylase (tgt) from Helicobacter pylori. AB - The aroB gene from Helicobacter pylori strain P1 was cloned and further characterized by sequence analysis and by functional complementation of the aroB mutation in Escherichia coli. The aroB gene encodes the enzyme 3-dehydroquinate synthase which catalyzes one of the early steps in the shikimate pathway. This pathway, which creates aromatic molecules from sugar precursors, is present in prokaryotes, fungi and plants but is absent from mammalian cells. The predicted amino acid sequence of the H. pylori aroB gene product showed significant homology (30-40% identity and 50-60% similarity) to 3-dehydroquinate synthases from various other prokaryotes and eukaryotes. The single gene on a plasmid was biologically active in E. coli. It suppressed the specific phenotype of aroB mutants by restoring the shikimate pathway-dependent synthesis of aromatic amino acids and the production of the siderophore enterobactin. Two other reading frames were found adjacent to the aroB gene. The first, designated as orf1, had no significant homology to proteins and genes present in databases, whereas the second was found to share a significant degree of homology with the tgt gene encoding tRNA-guanine transglycosylase from a variety of other bacteria (40-50% identity and 60-70% similarity). The function of the tgt gene was confirmed by heterologous complementation. The gene on a plasmid was shown to complement the queuosine biosynthesis defect in a genetically defined tgt- strain of E. coli. The presence of the aroB gene and the putative tgt homologue in unrelated H. pylori strains was confirmed by Southern blot hybridization and by polymerase chain reaction with specific primers. PMID- 9403841 TI - Interstrain variation of immediate early DNA sequences and glycoprotein B genotypes in cytomegalovirus clinical isolates. AB - Cytomegalovirus (CMV) disease is associated with a high mortality in recipients of an allogeneic stem cell transplant. Apparent differences in biological behaviour have been noted among clinical CMV isolates. By amplifying specific functionally relevant regions of the CMV genome [immediate early (IE) exon 3, glycoprotein B (gB)], a possible association of strain variation and clinical symptoms of infection was analysed in 24 patients. A high number of genome mutations of the IE exon 3 region could be documented translating into amino acid changes of viral isolated of 8 out of 15 patients with symptomatic and 2 out of 9 patients with asymptomatic CMV infection. Identical IE mutations and gB types were observed in isolates from two different sites in 6 patients. gB strain 2 was found to be associated with symptomatic CMV infection (P = 0.03). Thus, apart from host factors viral factors might influence the virus-host interaction in severely immunosuppressed patients. PMID- 9403842 TI - Differential uptake and killing potential of Campylobacter jejuni by human peripheral monocytes/macrophages. AB - The ability of Campylobacter jejuni to survive in monocytes after phagocytic uptake was tested in a new in vitro model using adherent macrophages derived from human peripheral monocytes. The cells were stimulated with cytokines before use to ensure full phagocytic and killing activity. The kinetics of uptake and killing of bacteria was followed for 72 h with 16 strains, including stool and blood isolates and laboratory adapted strains. Significant bacterial strain differences were not observed, but the viability of phagocytosed bacteria was dependent on the individual donating the macrophages. The majority of blood donors carried macrophages that killed phagocytosed Campylobacter within 24 or 48 h. There was no correlation between the source of isolation of the strains and relative intracellular survival. Bacterial mutants of superoxide dismutase, catalase or polyphosphate kinase were all as sensitive to macrophage killing as their isogenic wild-type strain. In contrast, about 10% of the voluntary blood donors carried monocytes which were incapable of killing phagocytosed bacteria. Such macrophages displayed normal uptake, but killing was insufficient and bacterial growth was observed with all strains and mutants tested. We conclude that (1) since in most cases activated human macrophages kill C. jejuni efficiently after phagocytosis, intra-phagocytic survival is not a common phenomenon during Campylobacter infection; and (2) those individuals carrying macrophages that are unable to destroy phagocytosed bacteria are at risk to develop a bacteremia during Campylobacter infection. PMID- 9403843 TI - Enzyme-linked immunosorbent assays with recombinant internal flagellin fragments derived from different species of Borrelia burgdorferi sensu lato for the serodiagnosis of Lyme neuroborreliosis. AB - The serodiagnosis of early Lyme neuroborreliosis is hampered by false negative results and one of the reasons could be the heterogeneity of strains of Borrelia burgdorferi sensu lato. For this study IgG enzyme-linked immunosorbent assays (ELISAs) with recombinant internal flagellin fragments (p41/i; amino acids 129 251) derived from strains PKo (B. afzelii), B31 (B. burgdorferi sensu stricto), and PBi (B. garinii) and ELISAs with whole-cell detergent extracts of strains PKo, PKa2 (B. burgdorferi sensu stricto), and PBi were compared. Cut off absorbance values were defined by the 94th and 92th percentiles of 200 sera from blood donors. Sera from patients with clinically defined neuroborreliosis [n = 88; 41 culture proven and 47 intentionally selected by the criteria specific IgG cerebrospinal fluid/serum index > or = 2 and a serum IgG immunofluorescence assay value of < 1:64] were tested. The sensitivities of the three whole-cell detergent extract ELISAs were similar (46.6-49.2%), whereas those of the recombinant ELISAs were highest with p41/i:PBi (57.1%) and lowest with p41/i:PKo (21.5%). A combination of the p41/i:PBi and the p41/i:B31 test was most sensitive (59.8%). The correlation of absorbance values of different assays was very high for the three whole-cell detergent-extract ELISAs, whereas the absorbance values obtained with the three recombinant tests differed considerably. The greatest differences were observed between p41/i:PKo and any of the other two recombinant antigens. The differences in immune reactivity of patients sera (due to strain heterogeneity?) seems to have more influence on the results of an assay based on a single antigen than on a whole-cell-based test. PMID- 9403844 TI - Detection and preliminary characterization of circulating immune complexes in patients with Lyme disease. AB - To investigate whether circulating immune complexes can be used as a disease marker for assessment of the activity of Lyme disease and for monitoring patients response to treatment, we tested 104 sera from patients with different stages of Lyme disease using the C1q enzyme-linked immunosorbent assay (ELISA) and a modified Raji cell test. Among 62 sera of patients with clinically active disease 27 sera (43.5%) reacted positively in the C1q-ELISA and 21 sera (33.9%) positively in the Raji cell test. In contrast, serum circulating immune complexes were found in less than 10% of 42 sera after antibiotic treatment. Similar results were obtained by both tests in 35 cerebrospinal fluid samples from patients with neuroborreliosis. Most importantly, dot blot analysis revealed the presence of both Borrelia burgdorferi-specific antigen(s) and host-derived components in the isolated immune complexes from serum samples of patients with active Lyme disease. These results indicate that detection of circulating immune complexes may be an useful parameter for judging the activity of Lyme disease. Moreover, preliminary characterization of spirochete-specific immune complexes implies new pathophysiological aspects of Lyme disease. PMID- 9403845 TI - Functional characterization of an isogenic meningococcal alpha-2,3 sialyltransferase mutant: the role of lipooligosaccharide sialylation for serum resistance in serogroup B meningococci. AB - The neisserial alpha-2,3-sialyltransferase, which is encoded by the lst gene, terminally links sialic acid to the lacto-N-neotetraose residue of neisserial lipooligosaccharide (LOS). We used the recently published nucleotide sequence of the neisserial lst gene to construct an isogenic serogroup B meningococcal lst mutant by insertion of a kanamycin resistance gene. The resulting lst mutant expressed the unsialylated lacto-N-neotetraose structure. Using bactericidal assays and an infant rat model of meningococcal infection, we were able to demonstrate that lst mutation, in contrast to galE mutation, which results in a truncated LOS, or to siaD mutation, which results in loss of the capsule, neither had an effect on resistance to normal human serum, nor did it impair the ability of meningococci to spread systemically in the non-immune host. The lst mutant was serum resistant despite of the fact that the central factor of complement activation, C3b, was deposited on the lst mutant as efficiently as it was on the galE mutant. Thus, the terminal sialic acid residue linked to the wild-type LOS inhibited C3b deposition on the meningocuccus. However, in contrast to the galE mutant, where C3b deposition is promoted by IgM binding, the lst mutant's surface is not a target for IgM molecules. Thus, the lacto-N-neotetraose residue of neisserial LOS alone, without the presence of terminal sialic acid, is sufficient to block IgM epitopes either on the LOS itself, or on other surface molecules. Our data provide further insight into the complex interplay of capsular and LOS sialic acids in serogroup B meningococci with host effector mechanisms, and suggest that LOS sialylation in meningococci is of a less central importance as it is in gonococci. PMID- 9403846 TI - Adventures in pharmacology, aspirin, prostacyclin and nitric oxide. PMID- 9403847 TI - Gene therapy. PMID- 9403848 TI - Molecular bases of neuroprotection in brain ischemia. PMID- 9403850 TI - The role of nitric oxide in blood cell interaction. PMID- 9403849 TI - Regulation of endothelial NO synthase (NOS3) expression in situations of vascular injury. PMID- 9403851 TI - Regulation of the nitric oxide/cyclic GMP system in astroglial cells. PMID- 9403852 TI - Role of nitric oxide in gastric functions. PMID- 9403853 TI - Antisense oligonucleotides in the design of new therapeutic strategies. PMID- 9403854 TI - Computer-assisted drug design: current status and future trends. PMID- 9403855 TI - Combinatorial organic synthesis and the lead discovery process. PMID- 9403856 TI - The impact of automation on high-throughput screening. PMID- 9403857 TI - Information technology in drug R&D. PMID- 9403858 TI - Desensitization of NMDA receptors as a mechanism of neuroprotection. PMID- 9403859 TI - Neuroprotective effects of functional antagonists at the glycine site of the NMDA receptor. PMID- 9403860 TI - Presynaptic receptors in the control of glutamate release: from facilitation to inhibition. PMID- 9403861 TI - Molecular mechanisms controlling apoptotic cell death in the nervous system. PMID- 9403862 TI - Neuroprotective effect of neurotrophic factors in experimental models of neurodegenerative disorders. PMID- 9403863 TI - Mechanisms of worsening in progressing stroke: from the laboratory to the clinical setting. PMID- 9403864 TI - Expression of mouse RNase MRP RNA in human embryonic kidney 293 cells. AB - We report on the expression of mouse RNase MRP RNA in human embryonic kidney 293 cells upon DNA transfection. Stable cell lines were selected by cotransfection with a neor gene. Transcription of wild-type and deletion mutants of MRP RNA and ribonucleoprotein formation were assessed by RNase protection and immunoprecipitation experiments. Mouse MRP RNA as expressed in 293 cells readily associates with human proteins to form a chimeric Th ribonucleoprotein. 5' truncated MRP RNAs, however, failed to associate with Th antigen(s) and deletion of the 3' sequences of MRP RNA greatly reduced the expression in stable as well as in transient transfectants. PMID- 9403865 TI - Affinity chromatography of RNase inhibitor. AB - Of the reagents used in reverse transcription-PCR (RT-PCR), RNase Inhibitor is the most costly on a per assay basis. A simple method for purification of RNase Inhibitor from bovine liver is described. Approximately 40,000 units of RNase Inhibitor can be purified to homogeneity from 400 g of bovine liver within two days using inexpensive reagents. The method described employs (a) facile procedures to rapidly obtain a clarified liver extract, (b) affinity chromatography of RNase Inhibitor on RNase-A-Sepharose, and (c) batchwise concentration of the purified protein with a molecular sieve resin. PMID- 9403866 TI - High order DNA structure as inferred by optical fluorimetry and scanning calorimetry. AB - New quantitative insights on the native high order chromatin-DNA structure existing within interphase nuclei are obtained by monitoring the effects of two common well-characterized fixatives, glutaraldehyde and ethanol/acetic acid mixture, at the level of the intranuclear DNA distribution and structures. Reproducible distinct levels of DNA fluorescence intensity and their intranuclear distribution are apparent in unfixed and fixed thymocytes by using DAPI and quantitative optical microscopy based on a charge coupled device. The fluorescent histograms correlated with the calorimetric thermograms on the very same thymocytes fixed and unfixed, establish an unequivocal baseline for the different levels of structural organization of the chromatin within the intact nucleus; namely their number, DNA packing ratio and fiber diameter. A systematic comparison among all the numerous models, being so far proposed for the quinternary and quaternary levels of DNA folding, to identifies the rope or ribbon-like and the chromonema as the ones that best fit with the in situ distribution. PMID- 9403868 TI - Contribution of Electron Paramagnetic Resonance to the studies of hemoglobin: the nitrosylhemoglobin system. AB - Since the initial work of Ingram (8,10) Electron Paramagnetic Resonance contributed considerably to research in hemoglobins. Now, 40 years later we review some of the results of the application of EPR to nitrosylhemoglobin (HbNO), as an example of the diversity of information which this technique can provide. PMID- 9403867 TI - Receptor-like protein tyrosine phosphatases: alike and yet so different. AB - Reversible phosphorylation on tyrosine residues is an extremely rapid and powerful posttranslational modification that is used in signalling pathways for the regulation of cell growth and differentiation. Over the past several years an impressive number of receptor-like protein tyrosine phosphatase (RPTPase) family members have been identified by molecular cloning, and undoubtedly many more will follow. This review provides an overview of the molecular data that are available for the currently identified RPTPases and discusses their possible biological implications. PMID- 9403870 TI - Preventing and treating rhinitis medicamentosa. PMID- 9403871 TI - Common adult infectious skin conditions. AB - Infectious skin conditions of the adult patient that primary care providers may often encounter in practice include those of viral (herpes simplex, herpes zoster, verruca and condylomata, molluscum), fungal (candidiasis, dermatophyte infections) and ectoparasitic (scabies, pediculosis) origin. Correct diagnosis and treatment of these conditions can provide relief to the patient and prevent spread to household and sexual contacts. This article discusses the epidemiology, etiology, history, appearance, diagnosis, and treatment of these commonly encountered infectious skin conditions. The primary care provider can differentiate these skin conditions by history, appearance, and laboratory tests, and should be able to diagnose and provide cost-effective therapy and secondary prevention for them. The primary care provider should also be able to recognize those lesions that are harbingers of systemic diseases and appropriately refer for further management. PMID- 9403869 TI - Tissue matrix protein expression in human osteoblasts, osteosarcoma tumors, and osteosarcoma cell lines. AB - Treatment for osteosarcoma is problematic because there are no prognostic markers. Diagnosis is primarily limited to cytologic grading. Oncogenesis alters cell structure therefore osteoblast tissue matrix proteins (extracellular matrix, cytoskeletal, intermediate filament, and nuclear matrix proteins), components of the cell substructure, are candidates for osteosarcoma markers. Structural proteins of the extracellular matrix, e.g. the collagens, are useful for diagnosis but not for tumors that produce little osteoid. To identify principal cellular tissue matrix proteins that distinguish normal from transformed human osteoblasts, their expression in normal osteoblasts, two osteosarcoma cell lines, and three primary osteosarcoma tumors were compared. The tumors were graded as (i) intermediate, (ii) high, and (iii) high grade recurrent. The 1-D SDS/PAGE profiles of the major components of the nuclear matrix and intermediate filament fractions from normal osteoblasts did not vary with biopsy site, age, or sex of patients. These profiles included known cytoskeletal proteins and OB250, a approximately 250 kD protein(s) observed in the intermediate filament fraction. A loss of protein bands, including OB250, was observed in the osteosarcoma cell lines and tumors. The intermediate and high grade tumors exhibited nearly identical protein profiles including potential tumor-specific proteins and collagen, consistent with the presence of intracellular collagen fibers in osteosarcoma. A microsequence was obtained for OT25, a novel low molecular weight protein observed in osteosarcoma cell lines. Fibrinogen gamma-chain, a protein that mediates cell adhesion was recovered from the high grade recurrent tumor. PMID- 9403873 TI - Gastroesophageal reflux disease: a common condition in the elderly. AB - Gastroesophageal reflux disease (GERD) is a condition frequently seen in the primary care setting and is more common among the elderly. Its manifestations may mimic other disease processes, and therefore the practitioner must be able to determine the cause of the presenting symptoms and know when to refer the patient for further diagnostic testing. When the presentation is straightforward, the practitioner should initiate treatment for GERD. Management of GERD must include patient education regarding postural measures, dietary and lifestyle modifications, and pharmacologic interventions. PMID- 9403872 TI - Emergency contraception: preventing unintended pregnancy. AB - Approximately 60% of all pregnancies are unintended at the time of conception, either unwanted or mistimed. Emergency contraception (i.e., use of a birth control method after intercourse has occurred) is a preventive treatment that has been underutilized. Six brands of oral contraceptives have recently been found by the U.S. Food and Drug Administration to be safe and effective as emergency contraceptive pills. These pills have been shown to reduce the likelihood of pregnancy occurring by at least 74%. Treatment with emergency contraceptive pills should be initiated within 72 hours of unprotected intercourse. Adverse effects include nausea, vomiting headache, and a change in menstrual bleeding patterns. Postcoital insertion of an intrauterine device is also highly effective as a postcoital contraceptive, but only in a select group of patients at low risk for sexually transmitted diseases. Promotion of access to emergency contraception includes educating both patients and providers about the method and making emergency contraceptive pills available to patients even before the need arises. PMID- 9403874 TI - Promoting healthy sexuality: guidelines for the school-age child and adolescent. AB - The purpose of this article is to provide the clinician with practical information to promote the healthy sexuality of the school-age child through adolescence. Much of the current literature related to sexuality is problem focused; this article focuses on sexuality from a health promotion viewpoint. The development of a healthy sexuality is a life-long process with different issues and problems pertaining to each age group. Issues common to the school-age child and adolescent include changing physical characteristics and body image concerns. A set of guidelines has been developed that can be used in the clinical setting. In order to follow a holistic approach to health promotion with these age groups, the clinician must address sexuality. These guidelines will provide the information the clinician will need to initiate a discussion about sexuality. Patient education guidelines are also presented that will help the parent, child, and adolescent in sexual issues and understanding. Suggested reading materials about sexuality are also presented. PMID- 9403875 TI - Influenza: immunization/prophylaxis for children, adults, and older adults. CDC Advisory Committee on Immunization Practices. PMID- 9403876 TI - Atorvastatin: a new agent for hyperlipidemia. PMID- 9403877 TI - A ministry of food for Britain for 21st century? PMID- 9403878 TI - Prevalence of malnutrition and vitamin A deficiency in Nigerian preschool children subsisting on high intakes of carotenes. AB - The prevalence of malnutrition and vitamin A deficiency was determined in 204 preschool children of both sexes aged 3-57 months. The children were recruited from 2 rural communities of Atakumosa Local Government Area of Osun State in South West Nigeria. Dietary vitamin A intake was estimated from frequency of consumption of locally available vitamin A containing food items. Vitamin A status of the children was assessed from concentration of retinol in plasma. Nutritional status was assessed from height and weight compared with international reference standards. The results indicate widespread malnutrition among the children. The prevalence of stunting (low height for age) was 60.8% while prevalence of wasting (low weight for height) was 7.4% and of underweight (low weight for age) 27.5%. Dietary vitamin A intake appeared to be adequate in the children. Intake of vitamin A is predominantly from plant sources. At least 43% of the children consumed the carotene rich red palm oil 6 or more times per week in contrast to less than 1% who consumed eggs or milk for 6 or more times per week. Vitamin A deficiency was low in the children. Only 11.3% of the children had plasma retinol concentration < 0.70 mumol/L. The results indicate that childhood malnutrition of public health magnitude can coexist with adequate dietary vitamin A intakes or vitamin A status. PMID- 9403879 TI - Effect of early complementary feeding on nutritional status in term infants in rural Nigeria. AB - Breastfeeding is common in developing countries, but exclusive breastfeeding is rare, and complementary foods are introduced at an early age. The objective of the present study is to determine the effect of early complementary feeding on the nutritional status of infants. Weight-for-age (WA) indices have been determined for 82 infants 3 to 4 months old participating in a breastfeeding study. They included 42 that started complementary feeding early (before 2 months) and 40 that started later. Weight-for-age indices were significantly lower for the early group than for the later group (t = 3.00, p = 0.004). The prevalence of underweight (WA SD scores below -2.0) was 7.5% in the later complementary feeding group and 28.6% in the early group (chi(2) = 4.76, p = 0.0292). Severe underweight (WA SD scores below -3.0) prevalences were 0% and 14.3% respectively (p = 0.0259). Thus, poorer nutritional status was significantly associated with earlier complementary feeding. The results suggest that exclusive breastfeeding, (together with promotion of weaning education and growth monitoring) should be vigorously promoted in these rural communities. PMID- 9403880 TI - The influence of time on dietary data: differences in reported summer and winter food consumption. AB - A pilot study was conducted to test whether or not the time of data collection affected subjects' responses to a dietary questionnaire and could therefore bias the results of a study. A 117 item food frequency questionnaire was administered to 43 subjects--16 in the summer and 27 in the winter. The summer and winter consumption per person per month was calculated and compared. Of the 117 foods, 105 (89%) showed differences between summer and winter consumption. While differences in summer fruit and hot cereal might be expected, this study showed that many other foods not usually thought of as seasonal were seasonally consumed by the population tested. Since food consumption is affected by supply and demand factors, and the reported memory of food intake is biased toward the present, the time of data collection could greatly influence reported intake for a great number of foods. Recommendations for enhancing dietary studies are given. These recommendations include the development of a gold standard of consumption for each food in a given population, incorporating the elements of person, place and time into study design, building the rate and length of data collection into the study plan and using the gold standard and improved design to produce a new generation of dietary studies. PMID- 9403881 TI - Nutrition education interventions in a community setting: 2 Dutch examples. AB - This article presents the results of two studies among participants of two Dutch nutrition education interventions, i.e. a self-help program and a living room group session. Both interventions aim to reduce fat intake in the general population and were developed as part of the Dutch community health project 'Healthy Bergeyk'. The objectives of the studies were to assess number of participants, participant characteristics, use of the self-help program, participant satisfaction and self-reported effects. Telephone interviews and written questionnaires were completed by inhabitants after participating in the interventions. The results are encouraging and the interventions seem to be useful as part of community projects. PMID- 9403882 TI - Effects of smoking on serum levels of lipid peroxides and essential fat-soluble antioxidants. AB - The effect of smoking on serum levels of the fat-soluble essential antioxidants (beta-carotene, alpha-tocopherol, gamma-tocopherol, and retinol) and lipid peroxides was studied in 31 smokers and 38 nonsmokers. The study subjects were male student, at King Saud University, aged 18-26 years. Smokers had significantly higher serum levels of lipid peroxides and significantly lower serum concentration of beta-carotene than nonsmokers (P < 0.05). Mean serum concentrations of retinol, alpha-tocopherol, gamma-tocopherol, and total vitamin E were lower among smokers than those of nonsmokers, but the differences were not statistically significant. The lower level of beta-carotene among smokers is probably brought about by the destruction of this antioxidant during neutralization of free radicals present in cigarette smoke. It seems possible that, during such oxidative stress, the interaction among antioxidants and their relative levels in serum are a crucial determinant of the concentration of a particular antioxidant. Therefore, the lower mean levels of alpha-tocopherol and gamma-tocopherol observed in smokers serum may also be related to cigarette smoking. This study has given rise to some concern about the adequacy of dietary beta-carotene, vitamin A and vitamin E of Saudis. The combination of cigarette smoking and low dietary intakes of essential antioxidants may provoke damage by oxidants present in cigarette smoke. PMID- 9403883 TI - Human bodily health and the common good. Proposals for action by individuals and the Catholic Church. PMID- 9403884 TI - Corneal geometry reconstruction with the Keratron videokeratographer. AB - BACKGROUND: Color-coded corneal maps produced by computer-assisted videokeratographers (CAVKs) have become an indispensable tool for clinical understanding of corneal shape. However, Placido-based CAVKs are criticized as theoretically incapable of producing accurate corneal height information. PURPOSE: This paper describes how the Keratron (Optikon 2000, Rome, Italy) integrated design innovations to achieve accuracy and map use previously thought to be unobtainable. METHODS: The Keratron implemented a spherically unbiased surface reconstruction method that yields height, axial power, and instantaneous curvature without derivation of one quantity from another. Processing innovations resulted in sub-micron height accuracy. The Keratron includes axial power, instantaneous curvature, refractive maps, a height map (spherical offset), pupil edge detection, a "move axis" feature, process editing, indices, a contact lens program, and photorefractive keratectomy (PRK) simulation. CONCLUSIONS: The algorithms for surface reconstruction and the design solutions implemented in the Keratron resulted in accurate height, axial power, and instantaneous curvature measurement and valid, clinically useful maps, as well as additional user options and features. PMID- 9403885 TI - Modeling the cornea with the topographic modeling system videokeratoscope. AB - The algorithms and models used by the Topographic Modeling System (TMS) corneal topography systems are described. The systems are calibrated to report the power at a number of points along each of 256 meridians. The power reported is the power of an equivalent sphere. From the radius of the equivalent sphere and the optical properties of the videokeratoscope, shape and a more accurate estimate of power are calculated. The TMS differs from most other videokeratoscopic systems in that it estimates the power and does not use axial distance. PMID- 9403886 TI - Reconstruction of the corneal shape with the MasterVue Corneal Topography System. AB - This paper explains how the Humphrey MasterVue Corneal Topography System measures the corneal shape and explains what is meant by the various values presented. Covered are the geometrical concepts underlying the method, the necessary reconstruction steps, a description of the arc step reconstruction method used, a description of the cone of focus concept, and definitions of the presented power values. PMID- 9403887 TI - Using corneal height maps and polynomial decomposition to determine corneal aberrations. AB - PURPOSE: To review the use of corneal videokeratoscopic height date, elaborate on the advantages and disadvantages of such data, describe techniques for overcoming the limitations of height data, and demonstrate its use in quantifying the optical properties and aberrations of the cornea. METHODS: The steep sag of the cornea hides fine variations in corneal height that arise naturally or due to disease or surgery. The dynamic range, or ratio of the overall sag to the feature height, is the primary limitation of videokeratoscopic height data. Techniques for removing single or multiple reference surfaces are described in detail, and applications of the methodology to wavefront and raytracing analysis of corneal aberrations arising from radial keratotomy (RK), photorefractive keratectomy (PRK), and keratoconus are described. RESULTS: Removing a single reference surface from the raw corneal height data begins to reveal subtle variations in corneal height. However, expansion of surface height data into a complete set of basis functions provides a sophisticated method for extracting high-order corneal variations. Choosing an orthogonal basis set provides a robust least-squares fit and forms unique expansions of the surface. The resulting coefficients are uncorrelated and form a simple measure of the optical quality. CONCLUSION: Videokeratoscopic height data are useful for analyzing and quantifying corneal deformity arising from disease or refractive surgery and they provide a sophisticated alternative or complement to dioptric power maps. PMID- 9403888 TI - Gaussian power with cylinder vector field representation for corneal topography maps. AB - The corneal maps displayed on commercially available corneal topography instruments are really one-dimensional, defining quantities only along a meridian, and ignoring shape information along any other direction. Both axial and instantaneous power have the drawbacks that there is a singularity at the center data point on the videokeratograph axis, and the computed powers and appearance of the corneal map change depending on the videokeratograph axis or on the location of asymmetries. We propose a new representation for corneal topography maps, Gaussian power with cylinder vector field, that has no singularity, and faithfully produces power values and corneal map patterns that are independent of videokeratograph axis or location of asymmetries. PMID- 9403889 TI - Mathematical model of a Placido disk keratometer and its implications for recovery of corneal topography. AB - The purpose of this paper is to illustrate the importance of radial contours in the target pattern of a Placido disk keratometer. We do so by presenting an example of a corneal surface which cannot be determined solely by the use of Placido ring images, but rather which requires radial contours for its determination. In order to prove our assertions, we derive partial differential equations (called the corneal transform), which relate the ring targets to their images. PMID- 9403890 TI - Axial curvature and the skew ray error in corneal topography. AB - BACKGROUND: Many intuitions about corneal shape are based on corneas with axial symmetry. In that case, the corneal normal intersects the axis of symmetry and there is an unambiguous definition for axial curvature. If, however, the corneal normal is not in the meridional plane, at least three definitions are available. The present paper defines, analyzes, and makes quantitative comparisons among the different definitions of axial curvature. When Placido rings are used as targets, current algorithms incur an error (the skew ray error) because the axial curvature that is measured is different from the axial curvature used for corneal reconstruction. METHODS: A cylindrical coordinate system is used that greatly simplifies the visualization and mathematics of skew rays. In the process of handling the axial curvature for different shapes, we develop a number of convenient formulas for handling ellipsoids and bispheres such as occur in keratoconus and photorefractive keratectomy (PRK). We also develop a simple approach based on vector cross-products for calculating axial curvature for an arbitrarily placed axis. RESULTS: The skew error is calculated for ellipsoidal and keratoconic corneas, and for corneas that have undergone decentered PRK ablation. For reasonable ellipsoidal and PRK corneas, the skew ray error is found to be negligible. However, for keratoconus and for decentered views of PRK corneas the error can be substantial. The implications of the skew ray ambiguity for instantaneous curvature are also discussed. CONCLUSIONS: The formalism developed in this paper facilitates the interpretation of axial curvature maps for corneas that are not axially symmetric. The skew ray error is negligible except for a small class of distorted corneas. PMID- 9403891 TI - Corneal topography reconstruction algorithm that avoids the skew ray ambiguity and the skew ray error. AB - BACKGROUND: Because corneal shape is typically measured with Placido ring targets, there is an ambiguity in determining which point on the ring corresponds to which point on the image. This ambiguity is expected to lead to errors (which we call the skew ray error) in reconstructing the corneal shape from Placido ring videokeratoscope (VKS) images. METHODS: A coordinate system based on cylindrical coordinates is developed for handling the case when the corneal normal does not lie in the meridional plane. A simple solution is found for the forward problem of locating the Placido ring object given the VKS image and the corneal slope. RESULTS: Algorithms for solving two inverse problems are developed: (1) the inverse problem of determining the Placido image given the Placido ring object and the corneal shape, and (2) the inverse problem of reconstructing the corneal shape based on knowledge of the Placido ring object and image. The algorithm compensates for the skew ray error and does not assume the cornea to be small. Both inverse algorithms are applied to a corneal shape with eight-fold corrugations similar to what might occur in radial keratotomy (RK). The reconstruction algorithm assumes sufficient smoothness for an arc-step algorithm to be possible. We find that the reconstruction algorithm does an excellent job of obtaining the correct corneal shape without the skew ray error. CONCLUSIONS: The concrete reconstruction algorithm demonstrates that one can recover the correct corneal shape just from the reflection of the Placido rings. A dart board pattern is not necessary for removing the skew ray ambiguity, as long as the cornea has sufficient smoothness in the radial direction, an assumption common to all reconstruction algorithms. PMID- 9403892 TI - Cone dimensions in keratoconus using Zernike polynomials. AB - PURPOSE: To determine the physical dimensions and location of the cone in keratoconic corneas from videokeratoscopic height data. METHODS: Corneal height date from keratoconus patients are obtained with a commercial videokeratoscope and decomposed into the set of orthogonal Zernike polynomials. The low-order Zernike terms are removed from the height data to expose the cone. The location of the peak of the cone and the cone's height and lateral dimensions are measured from the residual height data. This technique is compared to previous dioptric power-based methods of locating the cone. RESULTS: The corneal protrusion due to keratoconus takes on a variety of shapes and locations. These dimensions are easily measured with the Zernike decomposition technique and provide more detailed and accurate information regarding the cone than dioptric power data. CONCLUSIONS: The proposed method for quantifying cone dimensions and locale in keratoconic corneas provides practical and detailed information which may be useful in tracking the progression of the disease, contact lens fitting, and surgical planning. PMID- 9403893 TI - Cinemakeratography using computer morphing. AB - PURPOSE: TMS-1 videokeratography (Tomey Technologies, Inc.) was animated using computer morphing to visualize corneal shape change over time. METHODS: Morphing was performed using keratoconus (8 examinations/76 months) and postoperative photorefractive keratectomy (PRK) corneas (8 examinations/37 months; LSU PRK Phase IIa). RESULTS: In the keratoconus animation, the inferior cone increased curvature, but not equally in all directions, nor at a constant rate. The cone apex did not drift over time. Periodic flattening and steepening was observed superiorly, as well as cone regression at month 76. The Keratoconus Prediction Index (KPI) at month 0 was 0.18, plateaued at 0.36 at month 46, and dropped to 0.33 at month 76. In the post-PRK animation there was continuous corneal remodeling out to 30 months. Fluctuating steep and flat regions of the ablation zone correlated with the Surface Regularity Index (SRI). CONCLUSION: Cinemakeratography (Cinema-K) enhanced our appreciation of subtle changes in corneal topography. PMID- 9403894 TI - Care for postabortion complications: saving women's lives. AB - In developing countries each year more than half a million women die from maternal causes. Nearly all of these deaths could be prevented. Efforts to prevent maternal deaths from one major cause--complications of unsafe abortion- are crucial but inadequate in most of the world. Providing appropriate medical care immediately could save many thousands of women's lives. Offering family planning could prevent many future unintended pregnancies and unsafe abortions. PMID- 9403895 TI - Functional imaging and neuropsychiatry. PMID- 9403896 TI - Clinical and psychometric correlates of dopamine D2 binding in depression. AB - BACKGROUND: Single photon emission tomography (SPET) with the dopamine D2/3 ligand 123I-IBZM gives a semi-quantitative estimate of dopamine binding. In depressed patients, we predicted evidence of reduced function, i.e. increased binding, particularly in more retarded patients. METHODS: Fifteen depressed patients with major depressive illness and 15 healthy, age- and sex- matched volunteerS were examined with a clinical and neuropsychological test battery and high resolution IBZM-SPET. Estimates for specific binding were computed by averaging striatum to whole slice or frontal uptake ratios over 8-10 scans acquired from 70 min after tracer injection. RESULTS: Using whole slice as reference, left striatal uptake ratios did not significantly differ for patients from controls. Right ratios were significantly higher in patients than controls (P = 0.03). There were significant correlations between IBZM binding in left and right striatum and measures of reaction time and verbal fluency. CONCLUSIONS: Increased IBZM binding in striatum probably reflects reduced dopamine function, whether due to reduced release of dopamine, or secondary up-regulation of receptors. The observed abnormalities may be trait or state related, an issue that needs to be addressed with longitudinal study designs. The possible role of medication as a confounding variable requires further exploration. PMID- 9403897 TI - Structural brain abnormalities in male schizophrenics reflect fronto-temporal dissociation. AB - BACKGROUND: Many studies have separately reported abnormalities of frontal and temporal lobe structures in schizophrenia, but little is known of structural fronto-temporal associations in this condition. We investigated whether male patients with chronic schizophrenia would show abnormal patterns of correlation between regional brain volumes. METHODS: Structural magnetic resonance images of the brain in 42 patients were compared with 43 matched unaffected controls. We explored the pattern of association between regional brain volumes by correlational analyses, and non-parametrically tested for significance of between group differences by randomization. RESULTS: The schizophrenics demonstrated significant volume deficits in several brain regions (left temporal lobe and hippocampus, right dorsolateral prefrontal cortex), and significant volume increases in the ventricular system (third ventricle and left temporal horn of the lateral ventricle). Controls demonstrated large positive correlations (r > 0.4) between prefrontal and temporal lobe regions. By contrast, inter-regional correlations significantly reduced in schizophrenics included those between prefrontal, anterior cingulate and temporal regions, and between posterior cingulate and hippocampus (P < 0.05). The most salient abnormality in patients was a dissociation between prefrontal and superior temporal gyrus volumes (P < 0.01). CONCLUSIONS: These results support the existence of a relative 'fronto temporal dissociation' in schizophrenia which we suggest may be due to lack of mutually trophic influences during frontal and temporal lobe development. PMID- 9403898 TI - Temporal lobe magnetic resonance imaging can differentiate Alzheimer's disease from normal ageing, depression, vascular dementia and other causes of cognitive impairment. AB - BACKGROUND: Previous work suggests that temporal lobe magnetic resonance imaging (MRI) can distinguish those with dementia of the Alzheimer type (DAT) from healthy age-matched controls. However, its specificity with regard to conditions such as vascular dementia, depression and other disorders associated with cognitive impairment has not been determined. METHODS: We studied 222 subjects using T1 weighted MRI with 5.1 mm coronal slices throughout the temporal lobe. Subjects included: healthy controls (N = 40); DSM-III-R major depression (N = 61); NINCDS/ADRDA DAT (N = 77) and OTHER (N = 44, comprising subjects with vascular dementia, Huntington's disease, schizophrenia, alcohol related cognitive impairment and a group of 'memory complainers'). Hippocampus, amygdala, entorhinal cortex, parahippocampal gyrus and cerebral cortex were rated visually on a 0-3 scale by two experienced neuroradiologists blind to clinical diagnosis. RESULTS: Ratings of temporal lobe atrophy provided good separation between those with AD and all other groups. For example, anterior hippocampal atrophy had a sensitivity of 83% for detecting DAT, a specificity of 80% for controls, 87% for depressed subjects and 89% for OTHER. Other regions were less sensitive, but more specific for the diagnosis of DAT. In particular parahippocampal gyrus and entorhinal cortex had high specificity (97% for depressed subjects and 98% for OTHER). Because of an age-related increase in atrophy, sensitivity was highest for those over the age of 75, while specificity was highest for younger subjects. Significant correlations were observed between atrophy ratings of hippocampus, amygdala, entorhinal cortex and parahippocampal gyrus and CAMCOG memory score and length of history. CONCLUSIONS: Temporal lobe MRI may have an important role in assisting with the clinical diagnosis of DAT, particularly its differentiation from depression and other disorders that may cause diagnostic difficulties in clinical practice. PMID- 9403899 TI - Neuropsychological function in young patients with unipolar major depression. AB - BACKGROUND: While neuropsychological studies have consistently reported impaired cognition in elderly patients with unipolar depression, studies of cognitive function in younger patients with depression have produced equivocal results. The aim of this study was to examine the presence and nature of cognitive deficits in young patients with depression. METHODS: Neuropsychological function was assessed in 20 young patients with unipolar depression, in comparison to 20 age-, education- and IQ- matched controls. Subtests from the Cambridge Neuropsychological Test Automated Battery (CANTAB) were employed, as this battery has proved sensitive to deficits in middle-aged and elderly patients with depression. RESULTS: The patients were not impaired for short-term memory capacity, spatial working memory, planning ability, cognitive speed, delayed matching to sample or recognition memory. Compared to controls, the patients showed impaired subsequent movement latencies on the Tower of London task, suggesting deficits in the ability to sustain motor responses in depression. The depression group were also impaired on the task of attentional set shifting, requiring more trials to criterion at the intradimensional stage of the task and being more likely to fail the task at the extradimensional shift stage than controls. Further analysis indicated that half of the depression group failed to complete all stages of the set shifting task. These patients were more likely to have required in-patient hospitalization at some time during their illness. CONCLUSIONS: These results indicate that there are specific cognitive deficits in young patients with depression and that their presence may be related to a history of hospitalization. PMID- 9403900 TI - Use of a sequencing task designed to stress the supervisory system in schizophrenic subjects. AB - BACKGROUND: We investigated whether schizophrenic subjects are impaired in non routine behaviour because of the dysfunction of a general executive component labelled, in neuropsychological terms, the supervisory system. METHODS: A specific verbal sequencing test was designed for this purpose. Subjects had to perform sequential reasoning with verbal material. Each test sequence consisted of a series of words presented in jumbled order. The construction of some sequences had to be done using familiar routine associations (valid conditions). In contrast, some other sequences required the overriding selection of familiar routine associations, which were inappropriate within the general context of the task (invalid conditions). Twenty verbal sequences (10 valid-10 invalid) were administered. Thirty-seven DMS-IV schizophrenic patients and 21 normal volunteers matched for age and educational level were recruited. RESULTS: Compared to the control group the schizophrenic group was impaired in both valid and invalid conditions. The number of 'capture errors' specific to supervisory system failure was significantly higher in the schizophrenic group and only the schizophrenic patients had significantly fewer correct sequences in invalid conditions than in valid conditions. Poor performance in invalid conditions alone was observed only among the schizophrenic subjects without a general cognitive defect. CONCLUSIONS: These findings suggest that sequencing procedures requiring an executive input are impaired in schizophrenia. PMID- 9403901 TI - Heterogeneity in cognitive functioning of schizophrenic patients evaluated by a lexical decision task. AB - BACKGROUND: The disorganization pattern in schizophrenia, which involves formal thought disorders, is thought to be correlated with a deficit in integrative processes of contextual information. We tested the hypothesis that thought disordered schizophrenics, unlike non-though disordered schizophrenics, would present a deficit in the processing of the context during a task which involves these integrative processes. METHODS: A group of 22 schizophrenic patients diagnosed in accordance with DSM-III-R criteria and a group of 11 control subjects were compared using a semantic priming version of the lexical decision task. The experimental design used low-level structuration of verbal material to reveal the difficulty that schizophrenic patients encounter in using semantic regularities. RESULTS: A significant difference in priming effect was found between the three groups. Control subjects and non-thought disordered schizophrenics exhibit a priming effect for related word pairs when compared with unrelated pairs (respectively F(1,10) = 17.7; P < 0.002 and f(1,10) = 14.5; P > 0.003) but thought disordered schizophrenics did not (F(1,10) < 1; NS). CONCLUSIONS: This finding provides evidence for the cognitive heterogeneity of schizophrenic subjects. This absence of priming effect in thought-disordered schizophrenic subjects supports the hypothesis that these patients present a deficit in the post-lexical controlled information processing that permits the integration of semantic information. PMID- 9403902 TI - Psychopathology, executive (frontal) and general cognitive impairment in relation to duration of initially untreated versus subsequently treated psychosis in chronic schizophrenia. AB - BACKGROUND: It has been suggested that the expression of psychosis may reflect an active morbid process that is associated with increasingly poor outcome unless ameliorated by antipsychotic drugs. METHODS: The subjects of this study were 48 in-patients with schizophrenia, many of whom had been admitted before the introduction of antipsychotic drugs to rural Irish psychiatric hospitals in the late 1950s. Each patient was assessed for positive and negative symptoms, and for general and executive (frontal) cognitive function. RESULTS: After controlling for age and for duration and continuity of subsequent antipsychotic treatment, current severity both of negative symptoms and of general cognitive impairment was predicted strongly by increasing duration of initially untreated psychosis; duration of illness following initiation of antipsychotic medication failed to predict the severity thereof. Neither of these indices of illness duration predicted the severity of positive symptoms or of executive dyscontrol. CONCLUSIONS: Increasing duration of initially untreated psychosis was associated specifically with heightened accrual of prominent negative symptoms and general cognitive impairment. Executive dyscontrol, though also prominent in these patients, may be 'locked-in' at an earlier phase of the illness. PMID- 9403903 TI - IQ and risk for schizophrenia: a population-based cohort study. AB - BACKGROUND: This study aimed to quantify the association between low IQ and the later development of psychosis in a population-based cohort study of 18-year-old conscripts. METHODS: Fifty thousand males conscripted into the Swedish army in 1969-1970 were followed by means of the Swedish National Register of Psychiatric Care up to 1983. Tests of verbal and visuospatial abilities, general and mechanical knowledge and several psychosocial variables were recorded at conscription. RESULTS: One hundred and ninety-five subjects were admitted to hospital with schizophrenia and 192 with a non-schizophrenic psychosis on ICD-8 criteria. The distribution of scores in those later diagnosed as suffering from schizophrenia was shifted in a downward direction, with a linear relationship between low IQ and risk. This remained after adjustment for potential confounders. The risk for non-schizophrenic disorders was also higher in those with lower IQ but the effect was less marked and non-linear. Only poorer performance on the verbal tasks and mechanical knowledge test conferred a significantly increased risk for schizophrenia after taking into account general intellectual ability. Low IQ at conscription was not related to age of onset. CONCLUSIONS: The results confirm the importance of low intellectual ability as a risk factor for schizophrenia and other psychoses. This is unlikely to be due to prodromal decline or known confounders. The association could be directly causal with cognitive impairment leading to false beliefs and perceptions, or could be indirect with any factors causing lower IQ, such as abnormal brain development increasing the risk for schizophrenia. PMID- 9403904 TI - The care of patients with chronic schizophrenia: a comparison between two services. AB - BACKGROUND: There has been a widespread development of community multi disciplinary teams aimed to deliver coordinated comprehensive mental health care, yet there is little published evidence on the quality of care and economics of providing such care for people with severe mental illness. METHOD: This is a clustered randomized controlled economic comparison of the quality of care for patients with chronic schizophrenia by a multi-disciplinary community team with close links with primary care, and a traditional psychiatric service in a district general hospital psychiatric unit. RESULTS: Two years after it was established, patients with access to the community team had more of their needs met; they had fewer unmet needs; and they were more satisfied with the care they had received. They had more service contacts and received more interventions. The community team resulted in savings in the use of some hospital resources but these were not sufficient to offset the cost of the new service. The community team successfully directed care to patients with more needs, whereas no such relationship was evident for the traditional hospital-based service. Four years after the team was established, it met a greater proportion of needs for underactivity, daily living skills, use of public amenities and managing finances. CONCLUSIONS: Better quality care was provided at 2 and 4 years after its establishment by the multi-disciplinary community service than the traditional hospital-based service. Resources were targeted more efficiently by the community service. PMID- 9403905 TI - The effects of intelligence and education on the development of dementia. A test of the brain reserve hypothesis. AB - BACKGROUND: A number of recent epidemiological studies have shown that the prevalence and incidence of dementia are increased in population strata with low compared to high levels of education. This has been explained as a consequence of a greater 'brain reserve capacity' in people with a high level of education. Theoretically, however, brain reserve capacity is better reflected by intelligence than by level of education. Thus, the emergence of dementia will be better predicted by low pre-morbid intelligence than by low education. METHODS: This prediction was tested in a population based sample of elderly subjects (N = 2063; age range 65-84; Amsterdam Study of the Elderly) who were followed over 4 years. Dementia was diagnosed using the Geriatric Mental State examination (GMS). Pre-morbid intelligence was measured using the Dutch Adult Reading Test (DART), a short reading test which gives a good estimate of verbal intelligence, and is relatively insensitive to brain dysfunction. The effects of age, gender, occupational level, number of diseases affecting the central nervous system and family history of dementia or extreme forgetfulness were also examined. RESULTS: Logistic regression analysis showed that low DART-IQ predicted incident dementia better than low level of education. A high occupational level (having been in charge of subordinates) had a protective effect. CONCLUSIONS: This result supports the brain reserve theory. It also indicates that low pre-morbid intelligence is an important risk factor for cognitive decline and dementia. Use of reading ability tests is to be preferred over years of education as estimator of pre-morbid cognitive level in (epidemiological) dementia research. PMID- 9403906 TI - The omnipotence of voices: testing the validity of a cognitive model. AB - BACKGROUND: A preliminary report by the authors suggested that the range of affect generated by voices (anger, fear, elation) was linked not to the form, content or topography of voice activity, but to the beliefs patients held about them, in particular their supposed power and authority. We argued that this conformed to a cognitive model; that is, voice beliefs represent an attempt to understand the experience of voices, and cannot be understood by reference to the form/content of voices alone. This study puts this cognitive model to empirical test. METHODS: Sixty-two voice hearers conforming to ICD-10 schizophrenia or schizoaffective diagnoses were interviewed and completed standardized measures of voice activity; beliefs about voices and supporting evidence, coping behaviour; affect and depression. RESULTS: Beliefs about the power and meaning of voices showed a close relationship with coping behaviour and affect (malevolent voices were associated with fear and anger and were resisted; benevolent voices were associated with positive effect and were engaged) and accounted for the high rate of depression in the sample (53%). Measures of voice form and topography did not show any link with behaviour or affect and in only one-quarter of cases did neutral observers rate voice beliefs as 'following directly' from voice content. CONCLUSION: The study found support for our cognitive model and therapeutic approach. Factors governing the genesis of these key beliefs remain unknown. A number of hypotheses are discussed, which centre around the possibility that voice beliefs develop as part of an adaptive process to the experience of voices, and are underpinned by core beliefs about the individuals self-worth and interpersonal schemata. PMID- 9403907 TI - Continuity of care for patients with schizophrenia and related disorders: a comparative south-Verona and Groningen case-register study. AB - BACKGROUND: It is widely believed that for the severely mentally ill continuity of care is essential to ensure a better outcome and prevent long-term hospitalization. However, not much progress has been made in the operationalization and measurement of this concept. We used two indicators to compare continuity of care of schizophrenic patients in two kinds of mental health systems. One is a community mental health system without the back-up of a mental hospital (South-Verona, Italy). The other is an institution-based system in which mental hospitals are still predominant (Groningen, The Netherlands). METHODS: The first indicator of continuity of care, readiness of aftercare, is the time from discharge from hospital to the first day- or out-patient contact. Survival analysis was applied to correct for censored observations. The second indicator, flexibility of care, is the use of combinations of in-, day- and out patient care during 2-year follow-up. RESULTS: More patients in South-Verona received community care within 2 weeks after discharge (71.5%), than in the Groningen register area (54.6%). The survival functions differed significantly. Cox regression analysis revealed that in both systems a contact before admission, the time between this contact and admission and the duration of the admission are predictors for aftercare. A higher percentage of patients made multiple service use (combinations of in-, day- and out-patient care) in South-Verona than in Groningen (62 v. 45%). CONCLUSIONS: Both indicators showed a higher continuity of care in the South-Verona system. PMID- 9403908 TI - Incidence of first onset alcoholism among Taiwanese aborigines. AB - BACKGROUND: An initial prevalence survey of mental disorders among 993 subjects aged 15 and above randomly drawn from four major Taiwanese aboriginal groups (the Atayal, Ami, Bunun and Paiwan) was conducted from 1986 to 1988. The incidence of alcoholism was investigated in a follow-up survey from 1990 to 1992. METHODS: Both surveys employed a semi-structured clinical interview with satisfactory reliability for case identification, and DSM-III-R as the diagnostic criteria for alcohol use disorders. The estimation of incidence rates of first onset alcoholism (alcohol abuse or dependence) was based on person-years at risk of 499 subjects who did not have any lifetime diagnosis of such morbidity at phase I. RESULTS: The follow-up rate was 99.6% and only four subjects among the survivors were not found. The age-standardized annual incidence rates of alcoholism ranged from 2.8 to 4.9% among the four aboriginal groups, and the rank order of rates was consistent with that of prevalences among these groups. The incidence rates of alcoholism were the highest among adolescents and young adults in men, and among the middle-aged in women. CONCLUSIONS: High rates of first onset alcoholism among the Taiwanese aborigines indicate an interaction of sociocultural and biological factors in the development of such morbidity. PMID- 9403909 TI - Relationship of seizure duration to antidepressant efficacy in electroconvulsive therapy. AB - BACKGROUND: A relationship between the anticonvulsant and antidepressant properties of electroconvulsive therapy (ECT) has been hypothesized. The goal of this study was to see whether the anticonvulsant effects of ECT could be measured in a clinical setting and whether there was any relationship between the anticonvulsant effects of ECT and the antidepressant response to it. METHODS: We examined the temporal relationship between change in seizure duration (as an index of anticonvulsant activity) and improvement in Hamilton Rating Scale for Depression scores in a retrospective sample of 114 depressed patients who received 145 courses of ECT. A linear mixed effects model was utilized for analysis so that the repeated measures nature of the data could be taken into account. RESULTS: Both seizure duration and depression scores decreased significantly through the course of ECT. However, no evidence was found for a relationship between decrease in seizure duration and clinical improvement as measured by Hamilton ratings. CONCLUSIONS: The process underlying the reduction in seizure duration through a course of ECT may not be related to antidepressant efficacy. PMID- 9403910 TI - Genetic and environmental contributions to alcohol dependence risk in a national twin sample: consistency of findings in women and men. AB - BACKGROUND: Genetic influences on alcoholism risk are well-documented in men, but uncertain in women. We tested for gender differences in genetic influences on, and risk-factors for, DSM-III-R alcohol dependence (AD). METHOD: Diagnostic follow-up interviews were conducted in 1992-3 by telephone with twins from an Australian twin panel first surveyed in 1980-82 (N = 5889 respondents). Data were analysed using logistic regression models. RESULTS: Significantly higher twin pair concordances were observed in MZ compared to DZ same-sex twin pairs in women and men, even when data were weighted to adjust for over-representation of well educated respondents, and for selective attrition. AD risk was increased in younger birth cohorts, in Catholic males or women reporting no religious affiliation, in those reporting a history of conduct disorder or major depression and in those with high Neuroticism, Social Non-conformity, Toughmindedness, Novelty-Seeking or (in women only) Extraversion scores; and decreased in 'Other Protestants', weekly church attenders, and university-educated males. Controlling for these variables, however, did not remove the significant association with having an alcoholic MZ co-twin, implying that much of the genetic influence on AD risk remained unexplained. No significant gender difference in the genetic variance in AD was found (64% heritability, 95% confidence interval 32-73%). CONCLUSIONS: Genetic risk-factors play as important a role in determining AD risk in women as in men. With the exception of certain sociocultural variables such as religious affiliation, the same personality, sociodemographic and axis I correlates of alcoholism risk are observed in women and men. PMID- 9403911 TI - Consequences of major and minor depression in later life: a study of disability, well-being and service utilization. AB - BACKGROUND: The consequences of major depression for disability, impaired well being and service utilization have been studied primarily in younger adults. In all age groups the consequences of minor depression are virtually unknown. In later life, the increased co-morbidity with physical illness may modify the consequences of depression, warranting special study of the elderly. With rising numbers of elderly people, excess service utilization by depressed elderly represents an increasingly important issue. METHODS: Based on a large, random community-based sample of older inhabitants of the Netherlands (55-85 years), the associations of major and minor depression with various indicators of disability, well-being and service utilization were assessed, controlling for potential confounding factors. Depression was diagnosed using a two-stage screening design. Diagnosis took place in all subjects with high depressive symptom levels and a random sample of those with low depressive symptom levels. The study sample consists of all participants to diagnostic interviews (N = 646). RESULTS: As in younger adults, associations of both major and minor depression with disability and well-being remained significant after controlling for chronic disease and functional limitations. Adequate treatment is often not administered, even in subjects with major depression. As the vast majority of those depressed were recently seen by their general practitioners, treatment could have been provided in most cases. Bivariate analyses show that major and minor depression are associated with an excess use of non-mental health services, underscoring the importance of recognition. In multivariate analyses the evidence of excess service utilization was less compelling. CONCLUSIONS: Both major and minor depression are consequential for well-being and disability, supporting efforts to improve the recognition and treatment in primary care. However, controlled trials are necessary to assess the impact this may have on service utilization. PMID- 9403912 TI - Smooth pursuit eye movements in schizophrenia and affective disorder. AB - BACKGROUND: Smooth pursuit eye movement (SPEM) dysfunction is considered to be a promising candidate for a biological marker for genetic vulnerability to schizophrenia. There are conflicting findings regarding the question of what is exactly dysfunctional in SPEM dysfunction and what component of eye movements is really specific to schizophrenia. The purpose of the current study was to help to clarify the nature of (SPEM) dysfunction and its specificity to schizophrenia. METHODS: Smooth pursuit eye movements of 43 schizophrenic patients, 34 patients with major depression and 42 normal controls were examined using high resolution infrared oculography. These groups were compared on several indices of oculomotor functioning (gain, different saccadic categories). RESULTS: Schizophrenics had a significantly higher catch-up saccade rate than depressed patients and normals. The percentage of subjects with an abnormally high catch-up saccade rate defined as beyond the mean plus 2 S.D. of the normal control group was significantly higher in schizophrenics (27.9%) than in depressed patients (8.8%) and normal controls (0%). Low gain and higher numbers of intrusive saccades tended to be more prevalent in both patient groups but did not distinguish schizophrenics from depressed patients. CONCLUSIONS: Low gain and high rates of intrusive saccades contribute to SPEM dysfunction in major depression. Abnormally high rates of catch-up saccades seem to be the oculomotor component in smooth pursuit, that is specific to schizophrenia. PMID- 9403913 TI - Psychological preparedness for trauma as a protective factor in survivors of torture. AB - BACKGROUND: Although much research has focused on mechanisms of traumatization and factors related to post-trauma psychological functioning in survivors of trauma, there have been few studies of survivors of torture despite the widespread practice of torture in the world. The aim of this study was to examine the role of 'psychological preparedness' for trauma in post-traumatic stress responses in survivors of torture. METHOD: Thirty-four torture survivors who had no history of political activity, commitment to a political cause or group, or expectations of arrest and torture were compared with 55 tortured political activists, using structured interviews and measures of anxiety, depression, and post-traumatic stress disorder. RESULTS: Compared with tortured political activists, tortured non-activists were subject to relatively less severe torture but showed higher levels of psychopathology. Less psychological preparedness related to greater perceived distress during torture and more severe psychological problems, explaining 4% of the variance in general psychopathology and 9% of the variance in post-traumatic stress disorder symptoms. CONCLUSIONS: The study findings lend support to the role of prior immunization to traumatic stress and to unpredictability and uncontrollability of stressors in the effects of traumatization. Further research aimed at identifying the behavioural and cognitive components of psychological preparedness that play a role in traumatization may provide useful insights into effective treatment strategies for survivors of torture. PMID- 9403914 TI - In menstrual cycle stage a confounder in population-based psychiatric research? AB - BACKGROUND: It has been suggested that a failure to control for point in the menstrual cycle can lead to biased results in assessing psychiatric symptoms among women since state affects associated with premenstrual symptoms may lead to unreliability of symptom reporting as well as an artificial elevation of symptom ratings. We examine these hypotheses and the extent to which they can account for gender differences in symptom scale scores of demoralization and enervation. METHODS: The data are derived from an epidemiological study of Jews born in Israel between 1949 and 1958. The symptom scale scores of 2265 men and 1769 women (368 premenstrual, 458 menstruation and 943 postmenstrual) were compared regarding reliability, homogeneity and mean score. RESULTS: There were no differences among the menstrual groups, or between the men and women, in reliability of their responses as measured by the alpha coefficient and the coefficient of variation. There were no significant differences among the female groups on mean symptom scale score. The mean scale scores for each female group were significantly higher than the mean scores for men. CONCLUSIONS: Our results suggest that menstrual cycle stage does not influence the reliability of reporting, the variability of response or mean symptom levels. However, our conclusions may not apply to studies of drug effects or clinical studies of premenstrual dysphoria. PMID- 9403915 TI - Recurrent personal memories during intoxication reported by patients with alcoholism. AB - BACKGROUND: Experimental studies have demonstrated that alcohol has state dependent effects on learning and memory. We, therefore, sought to determine if alcohol intoxication triggers selective retrieval of memories which could alter patterns of alcohol use. METHODS: Eighteen alcoholic patients were studied as well as a comparison group of 12 patients who abused cocaine, a drug not associated with memory state-dependence. Patients underwent a semi-structured interview to elicit information about recurrent personal memories experienced when intoxicated. Recurrent memories experienced during craving were also studied as a comparison condition. RESULTS: The prevalence of recurrent personal memories during intoxication was reported to be much higher for alcoholic patients compared with the cocaine-abusing patients. These experiences occurred more frequently than during craving, generally reflected prior disturbing events and were often reported to promote continued drinking. CONCLUSIONS: The association of recurrently experienced personal memories with intoxication in alcoholic patients suggests, but does not establish, pharmacological state-dependence. Further studies of this memory phenomenon are indicated. PMID- 9403916 TI - Conceptual obstacles to research progress in affective disorders. PMID- 9403917 TI - Depression in multiple sclerosis. AB - BACKGROUND: An association between multiple sclerosis (MS) and depression has been recognized for several decades and has attracted considerable attention in research. However, there are considerable gaps in the current state of knowledge. In this review, the literature concerned with: (1) the burden of depression in MS; (2) the etiology of depression in MS, and (3) the treatment of depression in MS are critically examined. METHOD: The literature review utilized Medline (1966 1996), and was supplemented by citations extracted from the papers originally uncovered. RESULTS: Numerous studies have identified elevated depressive symptom scores in MS patients relative to nonclinical and (some) clinical control groups. Furthermore, studies of depressive disorders have clearly documented elevated prevalence rates in MS samples. The literature does not identify any specific pattern of neurological involvement as being consistently associated with depressive symptoms or disorders. Psychosocial risk factors contribute to the etiology of depression in MS, but the relative importance of various risk factors is yet to be determined. A single randomized controlled clinical trial, and additional anecdotal evidence, suggests that antidepressant pharmacotherapy is effective for depressive disorders in MS. CONCLUSIONS: Future epidemiological studies should not restrict their evaluation of risk factors to those specific factors that are closely related to the disease process. In particular, future researchers should resist the temptation to focus too exclusively on neuropathology. Biological, psychological and social risk factors are all potentially important. Additional empirical efforts to refine the various treatment approaches would be a welcome addition to this literature. PMID- 9403918 TI - The benefit of an insight-oriented and experiential approach on panic and agoraphobia symptoms. Results of a controlled comparison of client-centered therapy alone and in combination with behavioral exposure. AB - BACKGROUND: It is a common view that psychodynamic treatment does not help much to ameliorate the symptoms of panic and agoraphobia. The effects of an insight oriented treatment on central anxiety symptoms are the subject of the present controlled study. METHODS: Forty patients with severe panic and agoraphobia were admitted to an inpatient anxiety treatment program. Most of the patients had been treated by pharmacological means unsuccessfully. The patients were randomly assigned to pure client-centered therapy or to additional behavioral exposure treatment. Client-centered and behavioral agoraphobia manuals were used. The patients were examined on admission, at discharge and at 3, 6, and 12 months follow-up for panic (Structured Clinical Interview for DSM III-R--SCID), anxiety (Hamilton Anxiety Scale), agoraphobia (SCID, Fear Survey Schedule), and depressive (Hamilton Depression Scale) symptoms. RESULTS: Both client-centered treatment and a combination with exposure treatment reduced panic, avoidance and depressive symptoms significantly. For a short period the combined treatment was superior in patients' coping actively with anxiety and improving agoraphobic symptoms. However, at 1-year follow-up there was no further difference in the reduction of anxiety and depressive symptoms. CONCLUSIONS: The results are discussed with regard to the combination of these forms of therapy and to widespread skepticism about the efficacy of insight-oriented treatment. PMID- 9403919 TI - Psychological predictors of glycemic change with relaxation training in non insulin-dependent diabetes mellitus. AB - BACKGROUND: Previous findings are unclear regarding the possible glycemic benefits of applying behavioral relaxation training in non-insulin-dependent diabetes mellitus (NIDDM). METHODS: Subjects with NIDDM were randomized to relaxation training (6 sessions of progressive muscle relaxation and imagery, n = 12) or control treatment (routine medical care, n = 10). Physiological measures were total glycosylated hemoglobin (GHb) and area under the 2-hour oral-glucose tolerance curve (AUC). Psychological measures of generalized distress, anxiety and daily stress were also administered. All subjects were assessed before and after the 8-week intervention, and again at 16 weeks of long-term follow-up. RESULTS: There were no postintervention group differences in physiological variables. Highly distressed subjects and those who rated their glucose as more stress responsive tended to practice relaxation less between sessions. Within the treated group only, lower preintervention stress responsivity was associated with greater improvement in GHb, and lower anxiety and distress levels predicted long term improvement in AUC. CONCLUSIONS: It remains unclear whether relaxation training produces glycemic benefits in NIDDM. Perhaps the least anxious and stress-responsive patients only benefit from group-based relaxation training, whereas anxious individuals require intensive individually administered interventions. PMID- 9403920 TI - Personality traits and metabolic control: a study in insulin-dependent diabetes mellitus patients. AB - BACKGROUND: In the present study the authors evaluated the relationship between personality traits (according to DSM-III-R) and poor metabolic control in an adult onset insulin-dependent diabetes mellitus sample (n = 77). METHODS: Personality traits were assessed with the Personality Diagnostic Questionnaire- Revised. Metabolic control was evaluated through glycosilated hemoglobin (HbA1c): poor metabolic control was defined as HbA1c > or = 9% (normal values < 6.0%). RESULTS: Principal Component Analysis revealed three personality profiles: 'Cluster A/C Mixed', 'Cluster B Dependent' and 'Cluster B Aggressive'. Oneway ANCOVA, using sex as covariate, revealed a significant association (p = 0.01) only between poor metabolic control and Cluster B Dependent profile. No correlation was found between HbA1c and the other profiles. CONCLUSION: These data suggest that a specific personality profile is associated with poor metabolic control. PMID- 9403921 TI - Mood states and Type A behavior in Japanese male patients with myocardial infarction. AB - BACKGROUND: In Japanese studies, job involvement, e.g., job-centered lifestyle has been thought to be a major component in Type A behavior (TAB) rather than hostility and anger. We examined the influence of TAB including job involvement such as job-centered lifestyle on mood states such as depression following cardiac attack among Japanese male patients with myocardial infarction (MI). METHODS: After the first attack, the Jenkins Survey Activity was administered to 46 male patients with acute MI. Five years later, 33 patients remained as subjects. The Profile of Mood States was administered to the 33 subjects. RESULTS: The subjects are prone to unstable mood states compared to healthy controls. The severity of depression was significantly correlated with the global TAB. Of the global TAB, the job involvement was closely associated with depression as well as other mood states such as anxiety and fatigue. CONCLUSION: TAB, in particular job involvement, may be a predictor for depression following the cardiac attack. PMID- 9403923 TI - Young survivors of myocardial infarction. PMID- 9403922 TI - A comparison of Japanese and American psychiatrists' attitudes towards patients wishing to die in the general hospital. AB - BACKGROUND: The attitudes of the psychiatrists in Japan and the US were compared in order to investigate their ideas on whether patients in general medical hospitals who have a desire to die should be allowed to do so or be assisted in this regard, and whether they require psychiatric evaluation and intervention, and the cultural influences on these attitudes. METHODS: Japanese and American general hospital psychiatrists' attitudes towards the reasonability of suicide, physician-assisted suicide, and removal of life supports under various medical and psychosocial situations were compared. Seventy-two American and 62 Japanese psychiatrists' data were collected using the Suicidal Attitudes Inventory. RESULTS: The majority of both American and Japanese psychiatrists agreed that there may be times when suicidal ideation or completed suicide in med-surg patients could be reasonable. Significantly more Japanese psychiatrists responded with some agreement to the reasonability of suicide when one is unable to fulfill social role expectations, and had more concern about causing suicidal ideation by informing terminal patients of their diagnosis. CONCLUSIONS: The results indicate that psychiatrists' attitudes towards the relationship of psychopathology with suicidal ideation, the effect of depression, and other cultural factors on the desire to die in the medically ill are issues that need better clarification among both the medical profession as well as within society. Looking at how other societies handle these matters may help to understand one's own approach to them. PMID- 9403924 TI - Social-emotional behavior of preschool-age children with and without developmental delays. AB - Differences in parent and teacher ratings of social-emotional behavior among young children with developmental delays and those without significant developmental problems were examined. Participants included 198 preschool-age children identified as having a developmental delay (DD group) and 198 preschool age children without significant developmental problems (Comparison group) who were matched to the DD group by age and gender, using a randomized block procedure. Parent and teacher perceptions of social-emotional behavior of the participants were assessed using the Preschool and Kindergarten Behavior Scale (PKBS), a social skills and problem behavior rating scale for the use with young children. PKBS scores were found to classify the participants into their respective groups with a substantial degree of accuracy. Statistically significant differences in social skills and problem behavior scores between the two groups were found, with the DD participants evidencing greater social skills deficits and problem behavior excesses than the Comparison group. Individuals in the DD group were found to be four to five times more likely to have significant social skills deficits and problem behavior excesses than individuals in the Comparison group. The critical social-emotional behaviors separating the two groups appeared to be social interaction and independence skills, and socially withdrawn and isolated behavior patterns. New validity evidence for the PKBS is discussed, as are future needs pertaining to research and clinical practice in the area of social-emotional behavior of young children with developmental delays. PMID- 9403925 TI - The psychopathological model of mental retardation: theoretical and therapeutic considerations. AB - Mental retardation has heterogenous elements including genetic-biological and environmental factors that occur in a complex relationship. One construction commonly utilized is the overlapping etiologies, pathogenesis, and symptomatology in a bio-psycho-social model framework. A new integrated bio-psycho-social model we call "universe line" provides the possibility of integrating data from different research areas, specially cognitive and psychopathologic indicators. However, further methodology to refine and experimentally verify that model need to be elaborated. Implications of this theoretical approach are discussed. PMID- 9403926 TI - Measuring problem behaviors in children with mental retardation: dimensions and predictors. AB - Scores from the Child Behavior Checklist (CBCL; Achenbach, 1991a) and the Client Development Evaluation Report (CDER; California Department of Developmental Services, 1980) for 67 children and adolescents with mental retardation were examined to evaluate the factorial validity of the instruments. Four factor analyses were conducted. The initial factor analysis of CBCL data failed to confirm the presence of the five first-order factors previously reported for the CBCL standardization sample (Achenbach, 1991b). Second, the higher-order factors of Externalizing and Internalizing behaviors, similar to the structure reported for the CBCL standardization sample (Achenbach, 1991b), were confirmed on the present sample. Third, the two CDER factors of Personal Maladaption and Social Maladaption, previously identified by Widaman, Gibbs, and Geary (1987), were also confirmed. Finally, a higher-order factor analysis of the two factor scores from the CBCL and two factor scores from the CDER was conducted to study the congruence between the CBCL Externalizing and CDER Social Maladaption dimensions, and between the CBCL Internalizing and CDER Personal Maladaption factors. Moderate levels of congruence were found. Next, child characteristics, including level of mental retardation, age, and four dimensions of adaptive behavior, were used as predictors of problem behavior. No child characteristics were significantly related to the CBCL Externalizing dimension, but child age and level of mental retardation were significant predictors of the CBCL Internalizing dimension. CDER Cognitive Competence predicted CDER Social Maladaption, and child age predicted CDER Personal Maladaption. The findings are discussed in relation to previous studies of problem behaviors of children and adolescents with mental retardation. PMID- 9403927 TI - Validation of the Lifestress Inventory for people with a mild intellectual disability. AB - The Subjective Stress Scale (SSS; Bramston & Bostock, 1994) was developed to measure stress in people with a mild intellectual disability. In previous research, the SSS was found to measure two broad dimensions of stress (a) a General Worry factor and (b) a factor that tapped concerns about Negative Interpersonal Relations (Bramston & Fogarty, 1995). The present study sought to continue this line of research by introducing a slightly modified form of the SSS, to be known as the Lifestress Inventory (LI) and examining the psychometric properties of the scale when administered to a new sample of 221 people with mild intellectual disabilities. Confirmatory factor analysis indicated that three underlying factors corresponding to General Worry, Negative Interpersonal Relations, and Coping were sufficient to account for the correlations among the items in the LI. Rasch analysis indicated some improvements to the scoring format for the LI and also showed that the most easily experienced stressors were associated with the Negative Interpersonal Relations dimension. The refinements introduced by the LI and the further demonstration that some of the broad stress dimensions identified in the general population can also be found in people with an intellectual disability represent important milestones for researchers interested in exploring reactions to stress among this population. PMID- 9403928 TI - Characteristics of stereotypic movement disorder and self-injurious behavior assessed with the Diagnostic Assessment for the Severely Handicapped (DASH-II). AB - The first experiment involved 143 individuals with severe and profound mental retardation. Individuals with Stereotypic Movement Disorder, Self-Injurious Behavior (SIB), and Stereotypic movement disorder with self-injurious behavior as assessed by the Diagnostic Assessment for the Severely Handicapped-II DASH-II were validated against Diagnostic and Statistical Manual of Mental Disorders, fourth edition (DSM-IV; American Psychiatric Association, 1994) criteria. In a second study DASH-II scores for 1480 individuals with severe and profound mental retardation were compared on demographic variables, core and associated features of each disorder. Characteristics of persons in each group were reviewed. Persons with profound mental retardation were more likely to evince stereotypies or self injury compared to their severely impaired counterparts. Also, those with stereotypies were more likely to present with Pervasive Developmental Disorder (PDD)/autism, organicity, and eating disorders, while persons with SIB were more likely to evince sleep, sexual, and eating disorders. PMID- 9403929 TI - The effect of stereotypies on adaptive skills as assessed with the DASH-II and Vineland Adaptive Behavior Scales. AB - The relationship of the Stereotypy subscale of the Diagnostic Assessment for the Severely Handicapped-II (DASH-II) to adaptive functioning was investigated. Differences in adaptive skills measured with the Vineland Adaptive Behavior Scales (VABS) for individuals scoring at or above the cutoff of the Stereotypy scale and below the cutoff of the scale were analyzed. Individuals with high stereotypy scores had significantly lower VABS scores. Implications of these findings for assessment and treatment are discussed. PMID- 9403930 TI - Site-specific binding of polymerase-containing particles of the Giardia lamblia double-stranded RNA virus to the viral plus-strand RNA. AB - The non-segmented, double-stranded RNA genome of the Giardia lamblia virus (GLV) contains two genes encoding the major capsid protein (gag) and a fusion of gag with the viral RNA-dependent RNA polymerase (pol). Computer analysis of the viral RNA genome revealed three putative stem-loop structures that were predicted to mediate replication, transcription and packaging of the GLV genomic RNA by binding to the pol domain of the virus-encoded fusion protein. To provide evidence of these postulated RNA/protein interactions, gel retardation assays were employed to examine the potential binding capacity of various viral RNA genome-related sequences to native GLV protein(s). Viral proteins were obtained by disrupting purified GLV particles under low-ionic-strength conditions. The resulting viral protein particles maintained their RNA polymerase activity in the presence of GLV genomic RNA and thus appeared to be suitable tools for the analyses of GLV-protein-mediated binding reactions. A 72-nt short single-stranded in vitro transcript containing a putative stem-loop structure predicted to participate in the packaging of GLV (+)-strand RNA bound specifically to the disrupted virus particles. RNAs containing modified motifs of this stem-loop structure failed to bind to the GLV capsid. PMID- 9403931 TI - Glycoprotein-mediated biological properties of a host range mutant of Junin virus. AB - The biological properties mediated by the main envelope glycoprotein (GP1) of a mouse-attenuated mutant of Junin virus, named Cl67, were investigated in comparison with its parental strain XJCl3. In contrast to the parental strain, this mutant was unable to multiply in primary cultures of mouse embryo fibroblasts (MEFs). Impairment of Cl67 multiplication was associated with a lack of virus binding to MEF, probably due to an altered interaction between GP1 and cellular receptors. Antigenic and immunogenic characterization of GP1, performed by neutralization assays, demonstrated that, under certain conditions, polyclonal and monoclonal antibodies exhibited differential affinity and specificity for each virus. Cl67-infected Vero cells showed a marked pH-dependent fusion capability, suggesting more efficient low pH triggering of fusion by mutant virus GP1 in comparison with the parental strain. PMID- 9403932 TI - Previous heat shock treatment inhibits Mayaro virus replication in human lung adenocarcinoma (A549) cells. AB - Human lung adenocarcinoma cells (A549) were submitted to mild or severe heat shock (42 degrees C or 44 degrees C) for 1 h, while another group of cells was double-heat-shocked (submitted to 42 degrees C for 1 h, returned to 37 degrees C for 3 h, then exposed to 44 degrees C for 1 h). After each heat treatment, the cells were infected with Mayaro virus for 24 h and incubated at 37 degrees C. The results showed that the double-heat-shocked thermotolerant cells exhibited a 10(4)-fold virus titre inhibition, despite the recovery of protein synthesis and original morphology 24 h post-infection. In contrast, cells submitted to mild or severe heat shock exhibited weaker inhibition of Mayaro virus titre (10(2)-fold). The mildly heat-shocked cells also presented a full recovery in protein synthesis, which was not observed in severely heat-shocked cells. These results indicate that exposure of A549 cells to a mild or to a double heat shock treatment before Mayaro virus infection induces an antiviral state. PMID- 9403933 TI - Decrease in viral load associated with topical dinitrochlorobenzene therapy in HIV disease. AB - Quantitative measurement of human immunodeficiency virus (HIV) RNA is being used to assess the efficacy of various treatment modalities in HIV disease. Topical dinitrochlorobenzene (DNCB) therapy has been associated with improved clinical and immunologic parameters in HIV-infected patients. We have now examined the effect of topical DNCB treatment on plasma HIV RNA levels in a small prospective study. Eight HIV-infected subjects had T-cell counts and plasma viral load measured prior to initiation of DNCB therapy and 3-4 months after starting treatment. Six patients who refused DNCB therapy served as simultaneous controls. The mean CD4 and CD8 T-cell counts did not change significantly in either group over the study period. In contrast, plasma HIV RNA levels decreased one-half to greater than one log in each DNCB-treated subject, and the decrease in viral copies was statistically significant (p = 0.006). In the control group, plasma HIV RNA levels increased significantly over the course of the study (p = 0.037). We conclude that topical DNCB therapy has the ability to lower viral load in HIV infected patients. The long-term effect on viral burden of this inexpensive, readily available therapeutic modality merits further investigation. PMID- 9403934 TI - Mitogenic effect and activation of HIV1 production in serotonin-treated peripheral blood mononuclear cells derived from infected patients. PMID- 9403935 TI - Inhibition of sandfly fever Sicilian virus (Phlebovirus) replication in vitro by antiviral compounds. AB - Sandfly fever Sicilian virus (SFSV) was used in our laboratory to screen antiviral substances active toward viruses of the Bunyaviridae family. Antiviral activity was estimated by the reduction of the cytopathic effect of SFSV on infected Vero cells. Cytotoxicity was evaluated by determining the inhibition of Trypan blue exclusion. The specificity of action of each tested compound was estimated by the selectivity index (CD50/ED50). Selectivity indices of human recombinant interferon-alpha (IFN alpha) (Roferon and Introna), iota-, kappa- and lambda- carrageenans, fucoidan and 6-azauridine were much higher than that of ribavirin, the only antiviral substance which has been previously investigated for its inhibitory effects on Phlebovirus infections. Other compounds showed significant antiviral activity: glycyrrhizin, suramin sodium, dextran sulphate and pentosan polysulphate. All these compounds caused a concentration-dependent reduction in the virus yield. Ribavirin, 6-azauridine and IFN alpha have been shown to inhibit a late step of the virus replicative cycle, whereas glycyrrhizin and suramin sodium were active at an early step and the sulphated polysaccharides inhibited adsorption of SFSV on the cells. The antiviral compounds selected in this study as specific inhibitors of in vitro replication of SFSV are promising candidates for the chemotherapy of haemorrhagic fevers caused by viruses of the Bunyaviridae family. The combination of IFN alpha and ribavirin, which showed a synergistic antiviral effect, should be evaluated for the treatment of these infections. PMID- 9403937 TI - Endoscopic sinonasal surgery in the management of primary headaches. AB - Primary headaches (migraine, cluster, tension-type) are common disorders thought to be unrelated to nasal and sinus abnormalities. We present data on 19 patients with refractory primary headaches in the absence of significant sinus symptoms. The majority of patients responded with decreased pain to office application of nasal anaesthesia. A high prevalence of sinonasal abnormalities was found on coronal CT scans. Seventy-nine per cent responded with either decreased pain severity or headache frequency after endoscopic sinonasal surgery. We discuss possible underlying mechanisms to explain these findings. PMID- 9403936 TI - Distribution of HCV genotypes in patients infected by different sources. AB - To determine the distribution of hepatitis C virus (HCV) genotypes in our population within the Basque country, 58 HCV-infected patients were analysed. The predominant HCV genotype among subjects infected by blood transfusions (n = 9) and surgical procedures (n = 14) was 1b (88 and 50%, respectively). On the other hand, genotype 3a was the most common among intravenous drug users (n = 16), with data statistically significant (p < 0.0001). There were 18 patients (31%) with unknown risk factors, twelve of whom (66.7%) were infected with genotype 1b. Patients infected with genotype 1b were older than patients infected with genotype 3a (p < 0.0001). There existed a relationship between the HCV genotype and the source of infection. PMID- 9403938 TI - Combined endoscopic and subciliary orbital decompression for thyroid-related compressive optic neuropathy. AB - Compressive optic neuropathy is a feared, although unusual, complication of thyroid-related orbitopathy. A variety of surgical approaches have been described to achieve orbital decompression and alleviate the hallmark apical orbital crowding of this condition. We describe a subciliary anterior orbitotomy approach to the floor combined with an endoscopic medial wall resection. The anterior orbitotomy allows removal of the bones of the orbital floor both medial and lateral to the canal of the infraorbital nerve. The anterior orbital floor is retained for globe support. This combined approach retains the low morbidity of the endoscopic operation while achieving increased apical medial orbital wall and orbital floor decompression. We describe two illustrative cases where this approach produced a dramatic improvement in visual function. The surgical refinements associated with this combined approach offer technical advantages over other operations in the treatment of thyroid-related compressive optic neuropathy. PMID- 9403939 TI - Effect of nasal lavage on nasal symptoms and physiology in wood industry workers. AB - Nasal complaints and impaired nasal physiology are common in various occupational environments. Saline lavage has been recommended as treatment but has not yet been sufficiently evaluated. In this cross-sectional study of 45 wood industry workers, a significant decrease in nasal symptoms (such as obstruction, posterior secretions, itching, irritation and sneezing) was seen after a 3-week treatment with Rhinomer, which contains de-ionized, sterilized, isotonic seawater. Nasal peak expiratory flow (NPEF), especially in workers with nasal complaints, and nasal mucociliary clearance also improved significantly. The treatment, according to participants, was simple to perform and there were only a few side effects. PMID- 9403941 TI - The incidence of sinusitis in patients with multiple sclerosis. AB - A retrospective study was performed to assess the incidence of sinus disease in patients with MS. The MRI scans of 108 patients referred to a regional Neurosciences Unit with a diagnosis of multiple sclerosis were examined. There were 71 females and 37 males with an age range of 22 to 67 years (mean: 39.7 years). The sagittal and axial images were reviewed and the degree of sinus disease noted. This was graded as absent, minimal, polypoid and pansinus. Fifty seven patients (53%) had disease, the most common sinus involved was the maxillary followed by the ethmoid, frontal and sphenoid. Thirty-six patients had bilateral disease affecting the ethmoid sinuses most commonly. Three patients had fluid levels and four patients had retention cysts. The incidence of sinus disease is higher than in some other studies of normal populations. PMID- 9403940 TI - Polypoid rhinosinusitis in patients with host defence deficiencies: cellular infiltration and disease severity. AB - Polypoid rhinosinusitis is a chronic inflammatory, mucosal disease. Eosinophils may play a key role in driving and maintaining this inflammation. Polyps in conditions associated with chronic infective rhinosinusitis--such as cystic fibrosis (CF) and primary ciliary dyskinesia--however have been described as neutrophilic. We compared cell counts in polyps from 55 patients with host defence deficiencies (HDD) to polyps from 50 patients without HDD. The CT-scan appearance was also compared to the cell counts in the HDD group. No difference was detected in the percentage of patients with eosinophils from either group. Significantly more patients in the HDD group had polyp neutrophils (p < 0.001). Non-HDD-patient polyps contain more eosinophils (p < 0.000) whilst HDD-patient polyps contained more neutrophils (p = 0.005) and plasma cells (p = 0.05). Significant correlation was found between the neutrophil count and the CT score (p = 0.012) and the mast-cell count and the CT score (p = 0.02). Eosinophils are present in HDD and non-HDD polyps. Whilst the degree of cellular infiltration may vary, to classify polyps as eosinophilic or neutrophilic may be a false distinction. PMID- 9403942 TI - Relationship between nasal nitric oxide concentration and nasal airway resistance. AB - In the present study the relationship between nasal nitric oxide (NO) concentration and nasal airway resistance (NAR) was investigated in healthy volunteers at rest. Endothelially derived NO is established as a potent vasodilator and as such may be involved in the regulation of the nasal vasculature. Nasal airway resistance is dependent upon the tone of the nasal vasculature. It is therefore suggested that NO may play a role in the regulation of nasal airway resistance. Nasal NO concentration and nasal airway resistance were measured in 123 healthy volunteers. Posterior rhinomanometry was used to obtain the total and unilateral nasal airway resistance. Nasal NO concentration was measured from both the left and the right nostrils, consecutively, during a 20-sec breath hold, using a chemiluminescence gas analyser. NO was measured by sealing a cannula into each nostril consecutively and drawing air through both nasal passages. The results demonstrated that there was no significant difference between the concentrations of NO from the left and the right nostrils (p = 0.7). This indicated that the sampling technique provide a measure of nasal NO which was independent of the side of the nose used for sampling. The mean (+/- s.d) NO concentration sampled from the left nostril was 1,145 +/- 367 ppb. The mean NO concentration sampled from the right nostril was 1,163 +/- 401 ppb. There was a highly significant correlation between the right and left measurements (rho, corrected for ties = 0.95, p < 0.0001). The mean total NAR (+/- s.d) was 0.25 +/- 0.06 Pa/cm3/s. The mean left NAR was 0.50 +/- 0.28 Pa/cm3/s, whilst the mean right NAR was 0.48 +/- 0.31 Pa/cm3/s. There was no significant correlation between total NAR and the left nasal NO concentration (rho = 0.10) or total NAR and right nasal NO concentration (rho = 0.05). Similarly, no correlation was found between the left or right unilateral NAR and left or right nasal NO concentration, respectively. The results of the present study on healthy volunteers demonstrate that the nasal concentration of NO is not related to the total NAR. However, the present study cannot eliminate the possibility that nasal NO may be involved in the regulation of unilateral NAR. PMID- 9403943 TI - A quantitative appraisal of change in nasal tip projection after open rhinoplasty. AB - Exact knowledge about the magnitude of the factors influencing nasal tip projection (NTP) as a result of surgery are scarce. This study focuses on NTP changes after primary rhinoplasty using open transcolumellar approach in combination with a columellar strut, while specifically addressing four different nasal tip surgery techniques. Measurements were taken from standardised pre- and post-operative profile photographs of 73 patients with a minimal follow-up of one year. No statistically significant differences in pre- and post-operative NTP could be noted for the total group of patients. Only the group of patients in whom a nasal tip graft was applied a mean increase in NTP could be measured. A comparison of selected data from our study to data of previous studies on NTP changes after endonasal rhinoplasty demonstrates the effectiveness of the columellar strut for maintenance of NTP. Overall, our findings testify to the versatility of cartilage-structuring techniques including columellar struts and tip grafts for NTP maintenance or increase. The quantitative effect of the open approach itself on NTP remains elusive and needs further study. PMID- 9403945 TI - Isolated sphenoid sinusitis. AB - Isolated sphenoid sinusitis is a relatively rare clinical entity and can cause severe complications. Diagnostic nasal endoscopy using Hopkins telescopes and coronal and axial paranasal-sinus CT made the diagnosis of the sphenoid sinus disease easier. Eight out of 221 patients with paranasal sinus infection refractory to medical treatment--and treated surgically at the 2nd ENT Clinic of Ankara Numune Hospital between 1990-1995--had isolated sphenoid sinus infection. The most common symptom was headache felt in the retro-orbital region. Surgical procedure was intranasal endoscopic approach to the sphenoid sinus. The symptoms of the patients with isolated sphenoid sinusitis were completely resolved after surgery. As the literature is reviewed, it is concluded that endoscopic approach to the sphenoid sinus disease is the most appropriate method of surgery in order to reduce intra-operative morbidity and mortality. PMID- 9403944 TI - Patterns of hospital attendance with epistaxis. AB - The age and sex distribution of epistaxis admissions to hospital was examined. A retrospective analysis of 6,885 patients admitted to hospitals in the whole of Wales was performed, over a period of five years. The findings were compared with data from the 1991 National Population Census for the same region, thus providing a more representative estimate of the behaviour of this disease. A clear age relationship is demonstrated, with the incidence of epistaxis increasing rapidly after the age of 40 years. The female-to-male ratio is also age dependent. In the group aged between 20 to 49 years twice as many males as females were admitted, where no sex difference was expected from the population data. This difference was not present in the group aged 50 years and over where the ratio was similar to that in the underlying population. There was a 1.6 fold difference between the sex ratios of the two groups (95% confidence intervals of 1.9 to 1.4; p < 0.0001). It is possible that the female pre-menopausal state may provide a significant protection from this disease. The mechanism for this is unknown, but may be secondary to a direct effect of oestrogen on the nasal mucosa or vasculature, or the healing of vessels in this region. Alternatively, this observation may simply be a reflection of protection the pre-menopausal state provides against cardiovascular disease in general. PMID- 9403946 TI - Isolated sphenoid sinus aspergillomas. AB - Fungi are more often than previously believed to be the causative organisms of paranasal sinusitis. Aspergillus, a fungus belonging to the Ascomycetes class, accounts for the majority of these infections, which affect not only debilitated patients but healthy people as well. There are two distinct clinical forms of Aspergillus sinusitis, invasive and non-invasive, and each of them is further divided in two subtypes. Isolated aspergillosis of the sphenoid sinus is a rare disease, which is usually misdiagnosed for a long time because of its varying symptomatology. In the present study, four cases of isolated sphenoid Aspergillus disease are described and the recent literature is reviewed. Physicians should be aware of this rare clinical entity, as in many cases early diagnosis and appropriate treatment provide the key to achieve favourable outcomes. PMID- 9403947 TI - Cholesterol granuloma of the maxillary sinus. A case report. PMID- 9403948 TI - Poverty and health in West Germany. AB - The relationship between poverty and several health-related characteristics in West Germany was investigated. Data were derived from the National and Regional Health Surveys conducted in West Germany from 1984 to 1992. 25,544 males and 25,719 females with German nationality aged 25-69 years were examined. Poverty was defined as a household income of 50% less than the mean for West Germany. Multiple logistic regression analysis was used to analyze the relationship between poverty and four health variables: individual health behavior, subjective assessment of health status, cardiovascular disease risk factors, and self reported prevalence of lifetime chronic diseases. 10.2% of males and 12.8% of females were classified as being below the poverty line. For most but not all health parameters, less favourable results were found for the segment of the population with a household income below the poverty line. The most striking poverty-related differences were observed for lack of regular sport activities, subjective health satisfaction, obesity and myocardial infarction/stroke. Significantly lower prevalence rates for study subjects below the poverty line were observed for hypercholesterolemia in females only. Allergic disorders were the only chronic diseases reported significantly less often in males and females below the poverty line. Poverty has strong effects on individual health status and the prevalence of chronic diseases. Due to the rising unemployment rates in Germany in the last years it is very likely that the strong negative consequences of poverty for health are increasing. PMID- 9403949 TI - [Is there a correlation between alcohol drinking and sexual risk taking? A discussion of conceptional aspects exemplified by HIV infected men with homosexual behavior]. AB - Does sexual risk behavior and alcohol consumption correlate or is sexual risk behavior due to alcohol consumption? These questions are controversially discussed in the literature. We investigated whether the two different methodological concepts (global association, e.g. not specific to the critical incident and situational association) cause these discrepancies in a sample of 64 HIV-infected gay men. There was no significant global association between alcohol consumption and sexual risk behavior. Using the situational approach, drinking was not related to sexual risk behavior. The combination of these two concepts may help to identify persons at risk. PMID- 9403950 TI - [Assessment of AIDS risks: effects of different scale ranges and preventive knowledge]. AB - In a sample of 552 respondents effects of different scale ranges and respondents' knowledge about AIDS for answering questions about AIDS-risks were examined. Knowledge was assessed with 12 items related to risks for HIV infections and preventive measures. AIDS hazards for different groups (prostitutes, fixers, hospital personnel, homosexuals, and haemophiliacs) had to be rated with five items, one for each group. Response variations were assessed in three parallel questionnaire versions with different ranges of risk assignments. Estimations had to be performed in comparison with the average population. Some groups were consistently rated to be at high risk, others were assigned low risks for becoming infected with the HIV-virus. The analysis of response patterns revealed that the three versions of numeric quantifiers determined the risk ratings only to some extent. The distribution patterns suggest that our subjects had an ordinal risk ordering in mind that might better be expressed in verbal terms. The knowledge about AIDS had no systematic effects on response behavior. PMID- 9403951 TI - [Use of the International Classification of Diseases for disability insurance in Switzerland: study of reproducibility]. AB - The International Classification of Diseases (ICD-10) was introduced in a pilot phase for coding of diagnoses in a sample of cases encountered in the Swiss Disability Insurance (SDI), as a substitute for the SDI codes, judged unsuitable for the description of disability. The goal of this study is to evaluate the reliability of use of these codes, by studying the coding reproductibility between the Medical Observation Centers (MOC) and the Evaluation and Coordination Center. A second goal was to evaluate the feasibility of coding by non physicians. Reliability of diagnoses was examined by level of precision: chapters, groups, categories and subcategories. The agreement between the two coding Centers varies from 78%, at the chapter level, to 29% at the most detailed level. This agreement is improved if taking into account the three main diagnoses for the comparison, reaching 91%. The results of the physician coder, concerning the precision and reliability of coding, are significantly superior to those of an economist coder and a medical secretary coder. To improve coding agreement, it is proposed to train coding professionals, provide coding tools, clarify which diagnosis is the main one, and seek a consensus for cases of systematic disagreement. The ideal level of precision should not be so high as to produce diagnostic codes with low reliability. It should, however, be precise enough to give a satisfactory description of the problems encountered. PMID- 9403952 TI - Duration of hospitalization during the first two years after AIDS diagnosis: a descriptive study. AB - Has there been a change in the duration of periods of hospitalization during the first two years after diagnosis of AIDS between patients diagnosed before 1988, compared with patients diagnosed since 1988? A cohort of 212 AIDS patients was studied. They were diagnosed before December 31, 1990 and were hospitalized between January 1, 1981 and March 31, 1993 in the University Hospital of Geneva, Switzerland. Overall, the duration of hospitalization did not seem to differ according to the year of AIDS diagnosis, though the more recently diagnosed patients were hospitalized with a more advanced level of immunosuppression. However, the pattern of hospitalization was slightly different. The periods of hospitalization for subjects diagnosed before 1988 were relatively longer soon after the AIDS diagnosis and at a late stage in the course of the disease, whereas for the more recent patients the lengths of hospital stays were more uniform during the whole course of the disease. PMID- 9403953 TI - Methods of correcting for multiple testing: operating characteristics. AB - We examine the operating characteristics of 17 methods for correcting p-values for multiple testing on synthetic data with known statistical properties. These methods are derived p-values only and not the raw data. With the test cases, we systematically varied the number of p-values, the proportion of false null hypotheses, the probability that a false null hypothesis would result in a p value less than 5 per cent and the degree of correlation between p-values. We examined the effect of each of these factors on family-wise and false negative error rates and compared the false negative error rates of methods with an acceptable family-wise error. Only four methods were not bettered in this comparison. Unfortunately, however, a uniformly best method of those examined does not exist. A suggested strategy for examining corrections uses a succession of methods that are increasingly lax in family-wise error. A computer program for these corrections is available. PMID- 9403954 TI - Some comments on frequently used multiple endpoint adjustment methods in clinical trials. AB - Confirmatory clinical trials often classify clinical response variables into primary and secondary endpoints. The presence of two or more primary endpoints in a clinical trial usually means that some adjustments of the observed p-values for multiplicity of tests may be required for the control of the type I error rate. In this paper, we discuss statistical concerns associated with some commonly used multiple endpoint adjustment procedures. We also present limited Monte Carlo simulation results to demonstrate the performance of selected p-value-based methods in protecting the type I error rate. PMID- 9403955 TI - Different disease rates in two populations: how much is due to differences in risk factors? AB - Two populations with different disease rates may differ in their risk factors for the disease. If so, it is desirable to know what proportion of the disease excess in the high-risk population is attributable to its greater exposure to the risk factors. This proportion has been called the relative attributable risk (RAR). A related measure is the adjusted relative risk (ARR), defined as the ratio of rates in high-risk to low-risk populations that would be observed if the distribution of risk factors in the high-risk population equalled that of the low risk population. We present methods for obtaining consistent estimates and asymptotic confidence intervals for both the RAR and the ARR using data from case control studies in the two populations. The methods are applied to the problem of estimating the differences in ovarian cancer incidence between U.S. white women (high-risk) and U.S. black women (low-risk) attributable to differences in reproductive risk factors. Simulations show that the methods perform well; however, when the true RAR is close to 0 or 1 or when sample sizes are small, RAR estimates may fall outside the unit interval. We discuss circumstances when the true RAR lies outside the unit interval; in such circumstances the ARR is easier to interpret. PMID- 9403957 TI - Large sample Bayesian inference on the parameters of the proportional hazard models. AB - This paper considers large sample Bayesian analysis of the proportional hazards model when interest is in inference on the parameters and estimation of the log relative risk for specified covariate vectors rather than on prediction of the survival function. We use a normal prior distribution for the parameters and make inferences based on the derived posterior distribution. The suggested approach is much simpler than alternative Bayesian analyses previously suggested for the proportional hazards models. Using simulated data we compare estimates obtained from the Bayesian analysis with those obtained from the full proportional hazards model and the reduced model after backwards elimination. We show that under a wider range of assumptions, the Bayesian analysis provides reduced estimation errors and improved rejection of noise variables. Finally, we illustrate the methodology using data from a large study of prognostic markers in breast cancer. PMID- 9403956 TI - Bayes estimates for immunological progression rates in HIV disease. AB - We develop Bayesian methods for calculating shrinkage estimates of immunological progression rates (for example, CD4 count decline rates) in populations of HIV infected patients. These methods make the assumption that decline of immunological markers may be modelled as approximately linear on some suitable chosen scale. They are applicable in situations where seroconversion times are unknown and follow-up of patients is variable, with some patients having only sparse measurements of immunological markers. Fitting of models is achieved by Gibbs sampling and CD4 count data from 603 members of the Edinburgh City Hospital Cohort with at least two CD4 determinations are analysed to provide an illustration. It is found that Bayesian shrinkage estimates for CD4 slopes on the square root scale are much more effective predictors of future CD4 counts than the least squares estimates, with respect to squared error loss. Of various shrinkage estimators considered, the most effective corresponds to the simplest model, which can also be fitted using SAS. A characterization of the pattern of CD4 loss in the Edinburgh cohort is obtained (mean rate of decline on root scale 1.61 per annum, standard deviation 1.03) and the effect of various covariates (sex, age, risk category and HLA antigen type) on immunological progression is considered. It is found that homosexual men in Edinburgh and patients with HLA haplotype A1B8DR3 experience significantly faster loss of CD4. PMID- 9403958 TI - An examination of methods for sample size recalculation during an experiment. AB - In designing experiments, investigators frequently can specify an important effect that they wish to detect with high power, without the ability to provide an equally certain assessment of the variance of the response. If the experiment is designed based on a guess of the variance, an under-powered study may result. To remedy this problem, there have been several procedures proposed that obtain estimates of the variance from the data as they accrue and then recalculate the sample size accordingly. One class of procedures is fully sequential in that it assesses after each response whether the current sample size yields the desired power based on the current estimate of the variance. This approach is efficient, but it is not practical or advisable in many situations. Another class of procedures involves only two or three stages of sampling and recalculates the sample size based on the observed variance at designated times, perhaps coinciding with interim efficacy analyses. The two-stage approach can result in substantial oversampling, but it is feasible in many situations, whereas the three-stage approach corrects the problem of oversampling, but is less feasible. We propose a procedure that aims to combine the advantages of both the fully sequential and the two-stage approaches. This quasi-sequential procedure involves only two stages of sampling and it applies to the stopping rule from the fully sequential procedure to data beyond the initial sample which we obtain via multiple imputation. We show through simulations that when the initial sample size is substantially less than the correct sample size, the mean squared error of the final sample size calculated from the quasi-sequential procedure can be considerably less than that from the two-stage procedure. We compare the distributions of these recalculated sample sizes and discuss our findings for alternative procedures, as well. PMID- 9403959 TI - A review of tests for detecting a monotone dose-response relationship with ordinal response data. AB - This tutorial reviews methods for testing independence between discrete levels of a dose and an ordered categorical response variable. The tests are designed to be powerful for cases in which the response improves monotonically as dosage level increases. First, we show how to apply some standard tests for doubly-ordered contingency tables. Then, we show how to construct tests as part of a model building strategy. Other topics discussed include generalizations to stratified data, small-sample methods, and sample size and power considerations. PMID- 9403960 TI - Survival analysis of randomized clinical trials adjusted for patients who switch treatments. PMID- 9403961 TI - Detection of overall space-time clustering in a non-uniformly distributed population. PMID- 9403962 TI - Cardiotoxin-like basic protein (CLBP) from Naja naja atra is not a cardiotoxin. PMID- 9403963 TI - Pharmacologic characterization of botulinum toxin for basic science and medicine. AB - The use of Botulinum neurotoxin (BoNT) is increasing in both clinical and basic science. Clinically, intramuscular injection of nanogram quantities of BoNT is fast becoming the treatment of choice for a spectrum of disorders including movement disorders such as torticollis, blepharospasm, Meige Disease, and hemifacial spasm (Borodic et al., 1991, 1994a; Jankovic and Brin, 1991; Clarke, 1992). Neuroscientists are using BoNTs as tools to develop a better understanding of the mechanisms underlying the neurotransmitter release process. Consequently, our ability to accurately and reliably quantify the biologic activity of botulinum toxin has become more important than ever. The accurate measurement of the pharmacologic activity of BoNTs has become somewhat problematic with the most significant problems occurring with the clinical use of the toxins. The biologic activity of BoNTs has been measured using a variety of techniques including assessment of whole animal responses to in vitro effects on neurotransmitter release. The purpose of this review is to examine the approaches employed to characterize, quantify and investigate the actions of the BoNTs and to provide a guide to aid investigators in determining which of these methods is most appropriate for their particular application or use. PMID- 9403964 TI - The use of hens' eggs as an alternative to the conventional in vivo rodent assay for antidotes to haemorrhagic venoms. AB - One of the tests used routinely for the preclinical assessment of antivenom efficacy is the WHO-approved rodent intradermal skin test for assessing neutralization of venom-induced haemorrhagic activity. This is a useful test as in many viperid venoms haemorrhage is considered to be the principal lethal (pathogenic) venom effect in envenomed humans. The main problems with such an assay are, first, the necessity of using large numbers of experimental rodents (rats or mice) in order to obtain statistically significant results and, second, that the test must result in pain for the animals during the 24 hr assay period. The present study compares the rodent assay with an alternative assay using venom, in both the presence and absence of antidote, applied to a filter paper disc and placed on the highly vascularized yolk sac membrane of chickens' eggs at an early developmental stage. This avoids sensitivity to pain as reflex arcs have not yet developed, and haemorrhage or neutralization/inhibition of haemorrhage can be easily recorded. Preliminary results showed a high level of correlation between the results of the two tests when used to assess the efficacy of an antidote. It is hoped that the new assay will reduce the need for pain-sensitive experimental animals in the future. PMID- 9403965 TI - Cytotoxicity of a low molecular weight fraction from Aloe vera (Aloe barbadensis Miller) gel. AB - The cytotoxicity of a low mol. wt fraction (LMWF) obtained from Aloe vera gel was determined by two different assays. Firstly, exposure of monolayers of chicken fibroblasts to LMWF induced disruption of intercellular junctions and detachment of individual cells from the bottom of the flask, with formation of cell-free gaps in the monolayer. Secondly, LMWF inhibited the production of reactive oxygen species by human polymorphonuclear leukocytes stimulated by zymosan, as followed by luminol-dependent chemiluminescence. The toxic activity of LMWF was compared to that of sodium dodecyl sulfate (a well-known toxic substance), aloe-emodin and aloin (an anthraquinone and its precursor present in Aloe vera cortex) using the chemilumescence assay, and was found to be of similar potency to these toxic substances on a weight-to-weight basis. These results confirm that Aloe vera gel contains toxic low mol. wt compounds, and every effort must be made to limit the amount of these toxins in the commercially prepared Aloe vera gel products. PMID- 9403966 TI - The RNA-N-glycosidase activity of Shiga-like toxin I: kinetic parameters of the native and activated toxin. AB - Shiga toxin and Shiga-like toxins are ribosome-inactivating proteins with RNA-N glycosidase activity which remove a specific adenine from 28S RNA. The toxins are composed of an A subunit non-covalently associated to a multimer of receptor binding B subunits. Near the COOH-terminus of the A subunit, a disulfide-bonded loop contains two trypsin-sensitive arginine residues. Proteolytic nicking at these sites, followed by reduction, removes from the A subunit the C-terminal end together with the associated B subunits. The requirement of such cleavage for biological activity of Shiga toxin and Shiga-like toxins has been recently questioned. The present paper reports the kinetic constants of the adenine release from highly purified Artemia salina ribosomes catalysed by Shiga-like toxin I and by its A subunit before and after treatment with trypsin, urea and dithiothreitol or urea and dithiothreitol alone. All reactions had approximately the same Km (1 microM). The Kcat was 0.6 min-1 for the untreated holotoxin and 6 min-1 for the isolated A subunit, respectively. The trypsin treatment increased 1000-fold the Kcat of the holotoxin (770 min-1) and 100-fold the Kcat of the A subunit (640 min-1). The same Kcat (693 min -1) was also observed when the A subunit was treated only with urea and dithiothreitol. Thus the full activity of Shiga-like toxin I required not only removal of the B subunits but also activation of the A subunit itself. Such activation could be largely induced in vitro by drastic loosening of the molecule induced by urea and dithiothreitol, but in vivo would probably require a proteolytic cleavage of the toxin. Inactivation of ribosomes by Shiga-like toxin I did not require sensitization of ribosomes by ATP and macromolecular cofactors present in postribosomal supernatants. PMID- 9403967 TI - Structural features of aminoquinolines necessary for antagonist activity against botulinum neurotoxin. AB - Certain aminoquinoline antimalarial compounds, such as chloroquine, antagonize the paralytic actions of botulinum neurotoxins (BoNT). These studies have been extended to determine the critical structural groups necessary for synthetic aminoquinolines to have antagonist activity against BoNT. Isolated mouse hemidiaphragms were maintained at 36 degrees C and indirectly stimulated; the resulting isometric twitch tensions were recorded as a measure of synaptic function. The muscles were exposed to the test compounds before being treated with a challenge concentration of BoNT (typically 0.2 nM of serotype A). The time to onset of 50% muscle paralysis due to BoNT was used to assess quantitatively the efficacy of the test compounds, which were then ranked on the basis of the concentrations necessary to delay paralysis by a specified time increment. Of the compounds tested, those having a 7-chloro-4-aminoquinoline configuration, similar to chloroquine (or the structurally similar 6-chloro-9-amino acridine group in quinacrine), were most effective. Truncation of the alkyl-amino-alkyl group from chloroquine and conversion of the 4-amino nitrogen to a primary amine did not significantly alter its effectiveness as a BoNT antagonist. However, the 6-chloro or 8-chloro- isomers of chloroquine were essentially ineffective. These results suggest that aminoquinolines antagonize the paralytic actions of BoNT through interaction with a selective, stereospecific site that is not well correlated with antimalarial activity. PMID- 9403968 TI - Neoplastic nodular formation in mouse liver induced by repeated intraperitoneal injections of microcystin-LR. AB - Neoplastic nodules were observed in mice liver treated with microcystin-LR (MCLR) by the intraperitoneal (i.p.) route over 28 weeks. After 100 i.p. injections of a sublethal dose (20 micrograms/kg) of MCLR, neoplastic nodules were observed without the use of an initiator. Multiple neoplastic nodules up to 5 mm in diameter were observed in the liver of mice in both groups, i.e. those injected 100 times i.p. and those injected 100 times with a 2 month withdrawal. The cysteine conjugate of MCLR was detected mainly in the affected livers. In contrast, when 80 micrograms/kg was orally administered 100 times, characteristic chronic injuries such as fibrous changes and nodule formation were not observed. PMID- 9403970 TI - Bibliography of toxinology. PMID- 9403969 TI - Scorpion envenomation in Merida, Venezuela. AB - This study involves a review of the records of 64 children referred to the University of los Andes Hospital with the diagnosis of scorpion envenomation. The patients were divided into group 1 (local manifestations, 42.4%) and group 2 (systemic manifestations, 7.8%). Seven of these patients had severe cardiorespiratory complications, came from distinct geographical zones and received antivenin. Our results indicate that poisonous and extremely dangerous scorpions predominate in certain geographical zones within the state of Merida, Venezuela. PMID- 9403971 TI - Comparative pharmacokinetics of doramectin and ivermectin in cattle. AB - Plasma pharmacokinetics were compared for 40 cattle dosed by subcutaneous injection with doramectin or ivermectin (200 micrograms kg-1), commercial formulations of doramectin or ivermectin, 20 cattle per product). Doramectin exhibited a similar peak plasma concentration to ivermectin (about 32 ng ml-1), but the time to Cmax was longer for doramectin (5.3 +/- 0.35 days) than for ivermectin (4.0 +/- 0.28 days). The area under the curve from time 0 to infinity post-injection was significantly higher (p < 0.001) for doramectin (511 +/- 16 ng day ml-1) than for ivermectin (361 +/- 17 ng day ml-1). This was explained by a lower clearance, a lower volume of distribution and, probably, a higher bioavailability of doramectin over ivermectin. It is concluded that the pharmacokinetic differences between doramectin and ivermectin may explain the longer duration of preventive efficacy of doramectin. PMID- 9403972 TI - Persistent efficacy of doramectin against experimental challenge with Ostertagia ostertagi in cattle. AB - Two studies were conducted in North America to evaluate the persistent efficacy of doramectin injectable solution against experimental challenge with infective larvae of Ostertagia ostertagi. In both studies, four groups of 10 randomly assigned calves, negative for trichostrongyle-type eggs on fecal examination, were treated subcutaneously in the midline of the neck with saline (1 ml 50 kg-1) on Day 0 or doramectin (200 micrograms kg-1 = 1 ml 50 kg-1) on Day 0, 7, or 14. Two additional calves from the same pool of animals were randomly assigned as larval-viability monitors and received no treatment. Beginning on Day 14 and continuing through Day 28, the 40 treated calves each were given approximately 1000 infective larvae of O. ostertagi by gavage daily; the two larval-viability monitors were inoculated in a similar manner with approximately 30,000 larvae as a single dose on Day 28. Animals were slaughtered on Day 42 in one study and on Days 42, 43, or 46 in the second. The abomasum from each calf was harvested and processed for worm recovery. A 2% aliquot of abomasal contents plus wash was examined for worm quantification and identification. Geometric mean O. ostertagi burdens were calculated from the log (O. ostertagi count + 1) and were used to estimate percentage reduction. In both studies, doramectin injectable solution was > or = 99.6% efficacious in reducing infection resulting from challenge with infective larvae of O. ostertagi for at least 21 days posttreatment; by 28 days posttreatment, efficacy was 87.3% in one study and 99.7% in the other. PMID- 9403973 TI - Duration of anthelmintic efficacy of doramectin and ivermectin injectable solutions against naturally acquired nematode infections of cattle. AB - A comparison of persistent efficacy of doramectin injectable (D) and ivermectin injectable (I) was investigated under field conditions with treated permanent principal (PP) and interval-grazed principal (IGP) calves. The experiment was initiated on October 13, 1992 (day 0). Cattle used were crossbred beef heifers of 185 kg average weight and 8 to 10 months old. By random allotment, 66 calves were divided into two groups of 15 PP-D and PP-I calves for each treatment and two groups of 15 IGP-D and IGP-I calves for each treatment. Three extra or replacement calves were allotted for each group. Permanent principal calves in three replicates of five cattle per treatment grazed continuously on nematode contaminated replicate pastures from day 0 to day 70. At 2-week intervals, i.e., days 0 to 14, 14 to 28, 28 to 42, 42 to 56 and 56 to 70, one IGP-D and one IGP-I calf was grazed with each of the respective PP-D and PP-I calf replicates and necropsied 21 days after removal from pasture. All respective PP calves and IGP calves were treated with doramectin at 200 micrograms kg-1 or ivermectin at 200 micrograms kg-1 by s.c. injection on day 0. After the day 0-14 interval, all IGP D calves had zero egg counts. From the day 14-28 interval through the next three grazing intervals, the arithmetic mean egg counts of IGP-D calves were 18, 90, 281 and 31; those of IGP-I calves were 30, 226, 74 and 185. This suggested a persistence effect of approximately 2 to 4 weeks. In PP-D calves, egg counts reached a mean maximum at day 56 of only five eggs per gram, while counts of PP-I calves reached a peak of 40 on day 42. From the day 14-28 interval and through all subsequent intervals, arithmetic mean total worm counts from IGP-I calves were 58 to 73% greater than those in IGP-D tracers. A maximal total worm count of 4159 was observed in IGP-D calves of the day 42-56 interval; total worm counts in IGP-I calves from the day 14-28 interval through the day 42-56 interval were: 5420, 6739 and 9979, respectively. Haemonchus and Cooperia were higher in prevalence than Ostertagia in both treatments. Results of PP-D egg counts and total worm burdens in IGP-I calves indicated a high level of doramectin persistent activity for approximately 4 to 5 weeks and an advantage over persistence activity of ivermectin. PMID- 9403974 TI - Nematode reinfection following treatment of cattle with doramectin and ivermectin. AB - A study was conducted to investigate reinfection with nematodes in calves following treatment with doramectin or ivermectin administered subcutaneously at a dose rate of 200 micrograms kg-1 of body weight under conditions of natural challenge. Thirty calves were allocated to three groups of 10 calves each based on body weight, sex, breed and pre-treatment faecal egg counts (FEC) after grazing together on a common pasture for three months. Treatments were doramectin, ivermectin and no treatment. Calves were returned to the same pasture for 56 days, placed on dry lot from days 56 to 63 and then necropsied over days 64-66. Faecal egg counts were done at days -1 and 0, then bi-weekly from day 14 to 56 and day 63. Mean FEC in control calves continued to rise throughout the grazing period. Trichostrongyle FEC were significantly (P < 0.05) greater in the control group compared to either treated group at each sample time following treatment. At necropsy, a total geometric mean of 19,847 nematodes per calf was recovered from untreated controls, of which eight genera were identified. The predominant nematode genera were Ostertagia (8749), Nematodirus (3702) and Cooperia (1927). In the ivermectin-treated calves, geometric mean worm burden was similar to that of the untreated controls: A total of 20,349 nematodes per calf was present including the genera Nematodirus (8633), Ostertagia (4700) and Cooperia (1740). In contrast, the geometric mean worm burden in doramectin treated calves was 12,173, including Ostertagia (4310), Cooperia (1141) and Nematodirus (1667). Doramectin was more effective than ivermectin in protecting calves from reinfection over a 56-day post-treatment grazing period under conditions of natural challenge as measured by accumulated mean worm burdens. PMID- 9403975 TI - Comparative persistent efficacy of doramectin, ivermectin and fenbendazole against natural nematode infections in cattle. AB - A study was conducted in Argentina, to investigate the period of protection of a single injection of doramectin administered subcutaneously (s.c.) at 200 micrograms kg-1 (1 ml/50 kg) compared with single treatments of ivermectin (200 micrograms kg-1 s.c.) and fenbendazole (5 mg kg-1 p.o.), against field infections of gastrointestinal parasites of cattle. Eighty-three animals were selected and ranked on the basis of serial fecal egg counts (e.p.g.'s). From this group, three animals were slaughtered before treatment and their lungs, abomasum, small and large intestines, were processed for parasite counts and identification. The remaining 80 animals were allocated in ranked groups of four to a control or one of three treated groups. Animals of the four groups were grazed together in the same pasture for the duration of the study. Treatments were administered on Day 0. Individual fecal samples were collected at weekly intervals for the first 49 days post-treatment and twice a week from Day 52 to Day 84 (end of study). At each collection day fecal samples were pooled for coprocultures. On Day 28 and 56, two animals from each group, previously identified on Day 0, were killed and their parasite burdens determined. The duration of protection of a single injection of doramectin was longer than ivermectin or fenbendazole treatment. On Day 56, the total number of parasites found in doramectin-treated animals was significantly (P < 0.05) lower than parasite burdens found in either ivermectin- or fenbendazole-treated animals. The longer persistent activity of doramectin was expressed by the lower number of adults and L4 stages of Ostertagia ostertagi. Data from this experiment demonstrated the limitations of using fecal egg counts to evaluate the persistent efficacy of anthelmintics. The duration of activity of doramectin was demonstrated more accurately by parasite counts in cattle from each group since decreasing e.p.g.'s were seen in non-medicated animals without changes in total parasite burdens. PMID- 9403976 TI - Therapeutic and protective efficacy of doramectin injectable against gastrointestinal nematodes in cattle in New Zealand: a comparison with moxidectin and ivermectin pour-on formulations. AB - Two similar studies were conducted in New Zealand to compare the therapeutic and persistent activity of doramectin injectable, moxidectin pour-on, ivermectin pour on and oxfendazole oral drench when administered to nematode-infected cattle which were then grazed on common pastures. On day 0 (treatment day), 40 cattle were weighed, faecal sampled and allocated on the basis of day--3 faecal egg counts (FEC) to four treatment groups. Cattle were then treated with either doramectin by subcutaneous (s.c.) injection, moxidectin and ivermectin by topical application, or oxfendazole orally using label-recommended dosages. Oxfendazole treatment served primarily as a control to monitor reinfection without persistent activity. Faecal samples for nematode egg counts and coprocultures for larval differentiation were collected six times between day 0 and day 56 and all cattle were reweighed on day 56. Doramectin reduced pretreatment FEC by 99.1% in the first study and by 100% in the second study when assessed at 14 days posttreatment. Corresponding reductions for moxidectin were 80.8% and 85.2%, for ivermectin 86.0% and 80% and oxfendazole 78.3% and 100%, respectively. Posttreatment rise in FEC indicated that reinfection of moxidectin-treated animals occurred at the same time as oxfendazole controls in both trials. Posttreatment rise in FEC with ivermectin pour-on was similar to moxidectin and oxfendazole in one study, but in the other study ivermectin pour-on delayed the rise by 14-21 days. The rise in FEC for doramectin was delayed for 14-21 days in one study and at least 21 days in the other. The better parasite control provided by doramectin resulted in greater weight gains for cattle over the 56-day period as compared to moxidectin pour-on, ivermectin pour-on and oxfendazole in both trials. Gains of doramectin treated cattle were significantly (p < 0.05) greater than those of ivermectin and moxidectin groups in one trial and the oxfendazole group only in the other. PMID- 9403977 TI - The effectiveness of a single treatment with doramectin or ivermectin in the control of gastrointestinal nematodes in grazing yearling stocker cattle. AB - Four studies were conducted to a similar experimental design in the U.S. to evaluate the effectiveness of doramectin injectable administered to yearling stocker cattle in the control of gastrointestinal nematodiasis over the subsequent grazing period. Studies were conducted in Wisconsin (WI) and Arkansas (AR) during the summer season. The other two studies were conducted in Georgia (GA) and Mississippi (MS) during the winter/spring season. Doramectin was compared with both ivermectin injectable and ivermectin pour-on in the WI study, with ivermectin injectable alone in the GA study and with ivermectin pour-on alone in the other two studies. At each study site, an area of permanent pasture previously grazed by parasitized animals was subdivided by fencing into equal pasture units each with its own water supply. A treatment designation (non medicated control, doramectin injectable, ivermectin injectable or ivermectin pour-on) was randomly assigned to each pasture unit. Weaned beef calves with confirmed gastrointestinal nematode infections were randomly allotted to a pasture unit and corresponding treatment group. Each treatment group consisted of three replicates of seven animals per pasture unit (total 21 animals) in the WI study, three replicates of four or six animals per pasture unit (total 16 animals) in the AR study, five replicates of six animals per pasture unit (total 30 animals) in the GA study and three replicates of 12 animals per pasture unit (total 36 animals) in the MS study. Treatments were 1% doramectin injectable solution, 1% ivermectin injectable solution, 0.5% ivermectin pour-on solution or non-medicated controls. The injectables were administered at a dose of 1 ml/50 kg body weight (200 micrograms doramectin or ivermectin/kg) by subcutaneous injection in the neck. Ivermectin pour-on solution was administered topically at a dose of 1 ml/10 kg body weight (500 micrograms ivermectin/kg). After receiving their prescribed treatment, animals were placed on their designated pasture unit where they remained for the entire grazing period (84-140 days). Fecal nematode egg counts and body weights were monitored at predetermined intervals throughout each study. Doramectin treatment reduced pretreatment egg counts by between 95 and 100% by 21 days post-treatment. Subsequent rises in egg output from exposure to infective pastures were delayed by two to four weeks resulting in substantial reductions in total egg deposition over the grazing period and, therefore, potential pasture recontamination. Doramectin treatment resulted in substantial average daily weight gain advantages (0.152-0.272 kg) over the grazing season compared to non-medicated controls. Advantages were statistically significant (P < 0.05) in three of the four studies. There were no significant differences (P > 0.05) in average daily gain between the doramectin and ivermectin injectable or ivermectin pour-on treated groups. PMID- 9403978 TI - A comparison of the efficacy of two treatments of doramectin injectable, ivermectin injectable and ivermectin pour-on against naturally acquired gastrointestinal nematode infections of cattle during a winter-spring grazing season. AB - Four groups of 18 crossbred beef steer calves (three replicates of six per group) were used to compare persistent efficacy of doramectin injectable, ivermectin injectable and ivermectin pour-on against naturally acquired infections of gastrointestinal nematodes during winter-spring grazing in Louisiana. The experiment was initiated on January 11. Treatments administered on Day 0 and again on April 5 (Day 84, 12-week interval) were: Group 1, untreated controls (CONT); Group 2, doramectin (DOR) at 200 micrograms/kg, s.c. injection; Group 3, ivermectin (IVM-INJ) at 200 micrograms/kg, s.c. injection; Group 4, ivermectin pour-on (IVM-PO) at 500 micrograms/kg, back midline. The cattle were weighed and fecal samples (for egg counts and for culture-larval identification) were collected at regular intervals throughout the 161 day experiment. In the interval between Day 0 and 84, arithmetic mean egg counts of the CONT group averaged about 890 eggs per gram, but then decreased markedly between Days 119 and 126, and remained at a lower plane for the remainder of the experiment. From Day 28 to 56, egg counts of the DOR group were consistently lower (P < 0.05) than those of controls and both IVM-treated groups. Egg counts of the DOR group were always lowest after the second treatment, but differed (P < 0.05) only from IVM-PO counts between Days 119 and 140 (35 and 56 days after the second treatment). Ostertagia was the predominant genus, followed by Cooperia in all four groups. Oesophagostomum, Trichostrongylus, Haemonchus, and Bunostomum were other genera identified. Bodyweights of the DOR group remained significantly greater (P < 0.05) than those of all other groups from Day 112 through the end of the experiment. Total gains for the CONT, DOR, IVM-INJ, and IVM-PO groups were 96, 159, 147, and 150 kg, respectively; treated groups were significantly (P < 0.05) greater than CONT, but differences among treated groups was not significant (P > 0.05). PMID- 9403979 TI - Evaluation of the therapeutic and protective efficacy of doramectin against psoroptic scabies in cattle. AB - Three studies were conducted to evaluate the therapeutic and protective efficacy of doramectin when given by injection at a dose of 200 micrograms/kg against induced Psoroptes otis infestations of cattle. The first study investigated therapeutic efficacy. Mite infestations were established on 15 test animals held in stanchions by transfer of material from infested donor calves. Test animals were then allotted on the basis of mite counts to a treatment group (10 animals) which received doramectin and a control group (5 animals) which received saline. Skin scrapings were collected for mite counts on the day before treatment and on days 7, 14, 21 and 28 after treatment. Efficacy assessed on the basis of the proportion of animals cured by day 28 was 100%. The second study was designed to determine the duration of protective efficacy. Forty-eight scabies-free heifers were allotted to a treated group of 32 which received doramectin, and a control group of 16 which remained untreated. These treatment groups were each divided into eight subgroups. Commencing on treatment day and continuing at weekly intervals for 7 weeks, a subgroup of animals from each treatment was placed in stanchions and challenged by transfer of material from infested donor calves. Skin scrapings for mite counts were collected 7 and 14 days later. Infestations were successfully established on all untreated control calves. Doramectin prevented the establishment of infestation for three weeks and significantly (P < 0.05) reduced infestation levels for an additional two weeks. The third study established the duration of residual protection conferred by doramectin and ivermectin under contact transmission. Ninety-six scabies-free heifers were divided into two equal treatment groups. Animals in one group received doramectin and animals in the other group received ivermectin at its recommended dose of 200 micrograms/kg by subcutaneous injection. Each treatment group was then divided into eight subgroups of six animals. Commencing on treatment day and continuing at weekly intervals for 7 weeks a subgroup of animals from each treatment was exposed to purposely infested seeder animals for one week. Three animals from each treatment subgroup were then placed in individual stanchions in which grooming was prevented and the other three were placed together in a pen where normal grooming behavior was permitted. Skin scrapings for mite counts were collected at weekly intervals for up to 4 weeks. Doramectin provided complete protection against infestation for five weeks compared to four weeks for ivermectin. These periods were not influenced by grooming behavior. PMID- 9403980 TI - Evaluation of the protective efficacy of doramectin against sucking lice of cattle. AB - The protective efficacy of doramectin against sucking lice was evaluated under natural challenge conditions in Canada. Two studies with a similar experimental design were conducted in sequence. In each study, two groups of louse-free cattle received either doramectin at a dose of 200 micrograms/kg by subcutaneous injection or no treatment and were then mixed with louse-infested animals for the winter housing period. All animals were examined for lice on treatment day and thereafter at weekly intervals for 13 weeks. In the preliminary study, acquisition of infestation in the face of a very mild challenge of Linognathus vituli was delayed by a mean period of 49 days in doramectin-treated animals compared with untreated controls. The difference in the delay between the groups was significant (P < or = 0.04). In the second study, test animals experienced a moderate challenge of a mixed infestation of L. vituli and Solenopotes capillatus. Acquisition of infestation was delayed by a mean period of 25.6 days in doramectin-treated animals compared with controls, the difference between the two groups being highly significant (P < or = 0.0001). These studies confirmed the protective efficacy of doramectin against sucking lice under natural challenge conditions showing that infestation is prevented for a period of about 4 weeks following administration of the drug at its recommended dose. PMID- 9403981 TI - Protective efficacy of doramectin and ivermectin against Cochliomyia hominivorax. AB - Two studies were conducted in Brazil using induced infestations of the New World screwworm, Cochliomyia hominivorax, to investigate: a) the comparative prophylactic efficacy of doramectin and ivermectin at their recommended use levels (200 micrograms kg-1 s.c.), and b) the duration of protection of a single injection of doramectin. In the comparative efficacy study, two groups of six animals each were treated with ivermectin or doramectin. Two hours after treatment, four incisions were made. Each incision was infested with 30 first instar C. hominivorax larvae and their status evaluated daily for 7 days post treatment (p.t.). Doramectin treatment was 100% effective in prevention of C. hominivorax infestations whereas ivermectin efficacy was incomplete. First instar larvae were eliminated in doramectin-treated calves by 48 h p.t., while in the ivermectin group, C. hominivorax developed in over 29% of the incisions. Healing began in wounds of doramectin-treated animals at 24 h p.t. and was in progress in 100% of all wounds at 2 days p.t., while 50% of ivermectin-treated calves showed incisions with active lesions. In the duration of protection study, 24 calves were allocated to six groups (T1-T6) of four animals each. Three groups (T1, T3 and T5) were treated with saline and three groups (T2, T4 and T6) with doramectin. Animals were infested as described previously according to the following schedule: T1 and T2 at day 14, T3 and T4 at day 21, and T5 and T6 at day 28 p.t. Incisions were evaluated daily for 8 days post-infestation. Screwworm infestations and viable third-instar larvae developed of all incisions of saline treated calves, while doramectin was 100% effective preventing development of C. hominivorax for 21 days p.t. and showed partial activity at 28 days p.t. PMID- 9403982 TI - Impact of expert testimony on the believability of repressed memories. AB - Research suggests that people question the believability of trial testimony based on an alleged victim's previously repressed memories. Participants read one of six scenarios depicting the trial of a man accused of sexually assaulting a young girl. The alleged victim either reported the assault immediately (child witness) or waited 20 years to report it (adult witness). In the adult witness condition, the woman's memory for the event had either been repressed until recently or had always been available, and expert testimony was offered on behalf of the defense, the prosecution, both, or neither. Regression analyses revealed that women perceived the accuser's testimony as more believable and the defendant's testimony as less believable than men did. Similarly, the belief in the accuser's testimony decreased and the belief in the defendant's testimony increased when the accuser was an adult in contrast to a child, and when the defense offered expert testimony in contrast to its absence. In addition, guilty verdicts were associated with higher levels of accuser believability, lower levels of defendant believability and testimony based on repressed memories in contrast to testimony based on memories that were never repressed. PMID- 9403983 TI - Comparison of beginning counselors' responses to lesbian vs. heterosexual partner abuse. AB - This study compared responses of masters and doctoral level counseling students to two domestic violence scenarios. Participants read a two paragraph description of a battering incident involving either a heterosexual or lesbian couple and then gave their impressions via a series of open and closed ended questions. Scenarios were identical save the manipulation of sexual partner as same or opposite sex. Experience and/or education with battered and/or gay/lesbian clients is also examined. Results indicated that subjects perceived the heterosexual battering incident as more violent than the lesbian battering incident and would be more likely to charge the male batterer than the female batterer with assault. Differences in treatment recommendations were made according to sexual orientation of the victim. Less than half of the respondents had coursework or practical experience pertaining to domestic violence and/or gay/lesbian concerns. PMID- 9403984 TI - The psychological impact of withholding disclosure of child sexual abuse. AB - Researchers have found equivocal results with respect to whether the disclosure of child sexual abuse is helpful or not. The threat of harm as well as the possibility of being humiliated, not believed, or blamed, render the disclosure of child sexual abuse difficult for some victims. Suppressing of traumatic events has been linked to negative health effects. The current study investigated the relationship between the inability to fully disclose the abuse and subsequent traumatic symptomatology. Questionnaires including the Trauma Symptom Checklist 40, the Child Sexual Experiences Questionnaire, and the Parental Support Scale were completed by 204 victims of child sexual abuse. Multiple regression analyses were performed using traumatic symptomatology as the dependent variable. The extent to which a victim wanted to tell about the abuse but held back from doing so and the severity of the abuse were related to adult symptomatology. Findings suggest that victims enduring more severe abuse are more likely to hold back from fully disclosing the abuse which is associated with more trauma-related symptoms. PMID- 9403985 TI - Social ecology and entitlements involved in battering by heterosexual college males: contributions of family and peers. AB - This study examined how the social ecological factors of family history and relationships with peers were associated with 193 college men's partner violence and attitudes regarding battering. Multivariate (path) analyses revealed that witnessing paternal battering in childhood was both directly and indirectly (through male peer variables and attitudes concerning battering) related to a man's violence toward female partners. Specifically, those men who reported witnessing paternal domestic violence as a child were more likely to associate with male peers who are abusive and who provide informational support for relationship violence. Associating with abusive male peers and receiving male peer informational support for battering were also related to perpetrating relationship violence. Of particular interest were the findings that after controlling for witnessing paternal battering, male peer informational support exerted a direct effect on the increased likelihood of using violence against female partners, and that, in the path model predicting battering ever, witnessing battering ceased to be a significant predictor of men's violence when peer and attitudinal variables were considered. Male peer-related variables also predicted men's increased beliefs of entitlement to abuse female partners, and the belief that battering is justified directly affected partner violence perpetrated. These results support the inclusion of the broader social ecology of the batterer in examinations of male partner violence. PMID- 9403986 TI - Health care utilization and history of trauma among women in a primary care setting. AB - Participants were 150 women seen consecutively by a female family physician in an HMO setting for nonemergent medical care. Each participant completed a questionnaire that explored three areas of trauma. Twelve months after the administration of the questionnaire, medical records of each participant were reviewed for several measures of health care utilization (i.e., number of telephone contacts, physician visits, ongoing prescriptions, acute prescriptions, specialist referrals). Age, education, and current marital status were unrelated to medical utilization. Participants' acknowledged history of physical and emotional abuse significantly correlated with most measures of health care utilization, whereas sexual abuse generally did not. The implications of these findings are discussed. PMID- 9403987 TI - Victimization and perpetration rates of violence in gay and lesbian relationships: gender issues explored. AB - This study explores gender differences in victimization and perpetration experiences of gays and lesbians in intimate relationships. A sample of 283 gays and lesbians reported on their experiences both as victims and perpetrators of gay/lesbian relationship violence by completing a modified version of the Conflict Tactics Scale (Straus, Gelles, & Steinmetz, 1980). General results indicate that 47.5% of lesbians and 29.7% of gays have been victimized by a same sex partner. Further, lesbians reported an overall perpetration rate of 38% compared to 21.8% for gay men. Other findings were as follows: (1) lesbians were more likely to be classified as victims and perpetrators of violence than gay men; (2) lesbians were more likely to report pushing or being pushed than gay men; (3) lesbians reported experiencing a greater number of different victimization and perpetration tactics than gay men; and finally, (4) when items were weighted to create an indicator of severity, no significant differences between lesbians and gay men were found. PMID- 9403988 TI - Synthesis of chromogenic substrates of alpha-amylases on a cyclodextrin basis. AB - One-pot acetylation and subsequent partial acetolysis of alpha-, beta- and gamma cyclodextrins resulted in crystalline peracetylated malto-hexaose, -heptaose, and -octaose, respectively. Prolonged acetolysis of beta-cyclodextrin gave a mixture of acetylated maltooligosaccharides, from which peracetylated malto-triose, tetraose, and -pentaose were isolated. The acetylated oligosaccharides were converted into alpha-acetobromo derivatives, and then transformed into 4 nitrophenyl and 2-chloro-4-nitrophenyl beta-glycosides. From the 4-nitrophenyl glycosides 4,6-O-benzylidene derivatives were prepared, which were used together with the free glycosides as substrates of porcine pancreatic alpha-amylase. PMID- 9403989 TI - Structural determination of the lipo-chitin oligosaccharide nodulation signals produced by Rhizobium fredii HH103. AB - Rhizobium fredii HH103 produces extracellular signal molecules that are able to induce deformation of root hairs and nodule organogenesis of soybean. This strain produces a large variety of nodulation factors, consisting of a linear backbone of GlcNAc with different degrees of polymerization, bearing on the non-reducing residue various different N-acyl residues. The reducing terminal residue is 2-O methylfucosylated at position 6. Several analogous molecules substituted with fucose were also detected. PMID- 9403990 TI - Structural analysis of the exopolysaccharide produced by Pediococcus damnosus 2.6. AB - The exopolysaccharide produced by a ropy strain of Pediococcus damnosus (2.6) in a semi-defined medium was found to be an homopolymer composed of D-glucose. On the basis of monosaccharide and methylation analysis, 1H, 13C, 1D and 2D NMR experiments the polysaccharide was shown to consist of repeating units with the following structure. [sequence: see text] PMID- 9403991 TI - Matrix-assisted laser desorption ionization mass spectrometry for the analysis of monosulfated oligosaccharides. AB - Sulfated oligosaccharides are an important class of compounds in the field of glycobiology. Mass spectrometric analysis of these molecules is challenging due to their readiness to dissociate in sample preparation and their tendency to fragment during ionization. Moreover, their presence in small quantity in biological systems poses additional problems. We report the development of a mass spectrometric method based on matrix-assisted laser desorption ionization (MALDI) in a time-lag focusing time-of-flight mass spectrometer for the analysis of monosulfated oligosaccharides. It is found that coumarin 120 is an excellent matrix for the analysis of monosulfated disaccharides, whereas the use of a mixture of coumarin 120 and 6-aza-2-thiothymine is very effective for the ionization of sulfated trisaccharides and tetrasaccharides including those containing N-acetylneuraminic acid. Molecular ions for a series of synthetic sulfo/sialo beta Gal(1-->3)GlcNAc and beta Gal(1-->4)GlcNAc structures can thus be observed with subpicomole detection sensitivity using a uniform microcrystal matrix/sample preparation procedure. It is demonstrated that, with this matrix formulation, the presence of a high amount of sodium chloride or sodium phosphate buffer, which is often the case for the HPLC fractionated samples, does not deteriorate the MALDI performance. The analysis of mixtures containing different types of oligosaccharides is also examined. It is found that different classes of oligosaccharides require different matrix preparation methods. PMID- 9403992 TI - Structural characterization of novel L-galactose-containing oligosaccharide subunits of jojoba seed xyloglucans. AB - Jojoba seed xyloglucan was shown to be a convenient source of biologically active xyloglucan oligosaccharides that contain both L- and D-galactosyl residues [E. Zablackis et al., Science, 272 (1996) 1808-1810]. Oligosaccharides were isolated by liquid chromatography of the mixture of oligosaccharides generated by treating jojoba seed xyloglucan with a beta-(1-->4)-endoglucanase. The purified oligosaccharides were reduced with NaBH4, converting them to oligoglycosyl alditol derivatives that were structurally characterized by a combination of mass spectrometry and 2-dimensional NMR spectroscopy. This analysis established that jojoba xyloglucan oligosaccharides contain the novel side-chain [alpha-L-Gal p-(1 ->2)-beta-D-Galp-(1-->2)-alpha-D-Xyl p-(1-->6)-], which is structurally homologous to the fucose-containing side-chain [alpha-L-Fucp-(1-->2)-beta-D-Galp (1-->2)-alpha-D-Xyl p-(1-->6)-] found in other biologically active xyloglucan oligosaccharides. PMID- 9403993 TI - Synthesis of ganglioside GM1 containing a thioglycosidic bond to its labeled ceramide(s). A facile synthesis starting from natural gangliosides. AB - Capitalizing on the readily available ganglioside, GM1, we have devised a simple synthesis of labeled GM1 analogues with sulfur in place of oxygen in their linkage to the ceramide residue (SGM1). The sugar moiety of ganglioside GM1 was released by ozonolysis and subsequent alkaline fragmentation in good yield. During acetylation of the ganglioside sugar, the carboxyl group of the sialic acid residue lactonized with the 2-hydroxyl group of the inner galactose moiety (galactose II). The resulting sialoyl-II2-lactone of pentadeca-O-acetyl monosialogangliotetraose could be readily transformed into the alpha-glycosyl bromide. Subsequent treatment of this glycosyl bromide with potassium thioacetate afforded the sialoyl-II2-lactone of tetradeca-O-acetyl-1-S-acetyl-1-thio-beta monosialogangliotetra ose. The latter could be condensed with (2R, 3R, 4E)-3-O benzoyl-2-dichloroacetamido-1-iodo-4-octadecen -3-ol in methanolic sodium acetate to afford a protected lyso-SGM1 derivative. One-step removal of the protecting groups under alkaline conditions gave beta-monosialogangliotetraosyl thiosphingosine. This lyso-SGM1 was converted into labeled analogues of SGM1 using the N-succinimidoyl derivative of radiocarbon-labeled octanoic and octadecanoic acid, respectively. Subsequent actions of GM1-beta-galactosidase, beta-hexosaminidase A, sialidase and again GM1-beta-galactosidase on these labeled analogues of SGM1 in the presence of taurodeoxycholate produced the respective analogues of GM2, GM3, lactosylceramide and glucosylceramide, respectively. PMID- 9403994 TI - The system of sulfated alpha-(1-->3)-linked D-mannans from the red seaweed Nothogenia fastigiata: structures, antiherpetic and anticoagulant properties. AB - Structural analysis of two xylomannans extracted from Nothogenia fastigiata was carried out. The results are consistent with the general pattern previously reported for other xylo-mannans of the same system, alpha-(1-->3)-linked D mannans 2- and 6-sulfated and having single stubs of beta-(1-->2)-linked D xylose, but one of the new samples contains a significant amount of 2,6 disulfated units. Both xylomannans studied are obtained as complexes with a beta D-(1-->3)-, alpha-L-(1-->4)-galactan and a beta-D-(1-->3)-, beta-D-(1-->4)-'mixed linkage' xylan co-existing in the seaweed, a fact that limits the accuracy of the data determined. The structures of the galactan and the xylan are similar to those previously informed for this seaweed. The antiviral activity against four different herpes simplex viral strains and the anticoagulant properties of all the xylo-mannans of the system are reported. PMID- 9403995 TI - Representation and searching of carbohydrate structures using graph-theoretic techniques. AB - This paper describes how the carbohydrate structures in the Complex Carbohydrate Structure Database (CCSD) can be represented by labelled graphs, in which the nodes and edges of a graph are used to denote the residues and the inter-residue linkages, respectively, of a carbohydrate. These graph representations are then searched with a subgraph-isomorphism algorithm. We describe the use of one such algorithm, that due to Ullmann, and demonstrate that it provides a very precise way of searching the structures in CCSD. We also describe the use of screening techniques that can eliminate many of the CCSD structures from the subgraph isomorphism search, with a consequent increase in the speed of the search. PMID- 9403996 TI - Novel alginate lyases from marine bacterium Alteromonas sp. strain H-4. AB - A bacterium Alteromonas sp. strain H-4 isolated from Laminaria fronds produced extra- and intra-cellular alginate lyases and utilized alginate as its sole carbon source. An extracellular alginate lyase was purified from the culture supernatant of the strain and its substrate specificity was characterized. The estimated molecular mass of the enzyme was 32 kDa and the isoelectric point was 4.7. Both polyM and polyG block degrading activities were observed using the substrate-containing gel overlay technique after isoelectric focusing of the enzyme. By analyzing the reaction products from the polyM block, polyG block, MG random block and intact alginate, three major peaks containing unsaturated tri uronide through octa-uronide were detected for each substrate. The results indicate that the enzyme of Alteromonas sp. H-4 can degrade both polyM and polyG blocks with a K(m) in mg/mL 20-times higher for the polyM block. PMID- 9403997 TI - Chemical characterization of a new 5,7-diamino-3,5,7,9-tetradeoxynonulosonic acid released by mild acid hydrolysis of the Legionella pneumophila serogroup 1 lipopolysaccharide. AB - A derivative of a new 5,7-diamino-3,5,7,9-tetradeoxynonulosonic acid was released from the lipopolysaccharide of Legionella pneumophila serogroup 1 (strain Philadelphia 1) by mild acid hydrolysis, and identified, using NMR spectroscopy and GLC-MS, as 5,7-diacetamido-8-O-acetyl-3,5,7,9-tetradeoxy-L-glycero-D-talo- nonulosonic acid or its enantiomer. PMID- 9403998 TI - Preparation, conformation, and mild hydrolysis of 1-glycosyl-2-acetylhydrazines of the hexoses, pentoses, 2-acetamido-2-deoxyhexoses, and fucose. AB - The title compounds were prepared and their conformations studied by 1H-NMR. Their acid hydrolysis under mild conditions was monitored by 1H-NMR. PMID- 9403999 TI - Identification of an alpha-D-Manp-(1-->8)-Kdo disaccharide in the inner core region and the structure of the complete core region of the Legionella pneumophila serogroup 1 lipopolysaccharide. AB - A disaccharide alpha-D-mannopyranosyl-(1-->8)-3-deoxy-D-manno-octulosonic acid [alpha-D-Manp-(1-->8)-Kdo] was released by mild acid degradation of Legionella pneumophila serogroup 1 (strain Philadelphia 1) lipopolysaccharide (LPS) and identified using NMR spectroscopy and GLC-MS of derived products. These data, together with methylation analysis of the native LPS and previously reported data [Y.A. Knirel, H. Moll, and U. Zahringer, Carbohydr. Res., 293 (1996) 223-234], allowed elucidation of the complete core region of the LPS as having the following nonasaccharide structure: [Sequence: see text] PMID- 9404000 TI - Stimulation of translation and cytoplasmic polyadenylation by the Xenopus c-mycI 3'-untranslated region. AB - Expression of messenger RNA (mRNA) in both embryonic and adult cells may be profoundly influenced by untranslated sequences in the 3'-end. Elements in the 3' untranslated regions (UTRs) of messengers are known to influence messenger stability, polyadenylation, and translation. We have examined the effects of the 3'-UTR of Xenopus laevis c-mycI (either alone or in combination with the 5'-first exon) on the expression of a chloramphenicol acetyltransferase (CAT) reporter in Xenopus embryos. The Xenopus c-mycI 3'-UTR enhanced messenger translation independent of the 5'-UTR. RNase H analysis indicated that the Xenopus c-mycI 3' UTR can promote the cytoplasmic polyadenylation of CAT mRNA in embryos. The result suggests that the post-fertilization enhancement of translation caused by the c-mycI 3'-UTR may be a consequence of cytoplasmic polyadenylation. A uridine (U)-rich sequence in the Xenopus c-mycI 3'-UTR that may be responsible for polyadenylation is similar to an element that destabilizes mammalian c-myc transcripts. We discuss the possibility that U-rich sequences may play a dual role by destabilizing growth-related transcripts in adult cells and stimulating their polyadenylation during development, and we propose that a switch in the role of such sequences in adult cells could lead to stabilization of these messengers, increased translation, and abnormal growth control. PMID- 9404001 TI - Predominant expression of a Z-chromosome-linked immunoglobulin superfamily gene, ZOV3, in steroidogenic cells of ovarian follicles and in embryonic gonads of chickens. AB - A cDNA clone, pZOV3, was isolated from the cDNA library of immature chicken ovaries and its gene was mapped to the middle of the short arm of the Z chromosome. The cDNA sequence suggests that ZOV3 is a novel member of the immunoglobulin superfamily. cDNA clones of homologues of chicken ZOV3 were also obtained from Japanese quail and pigeon. Northern blot hybridization suggests that the high-level expression of the ZOV3 gene is restricted to the gonads: embryonic, immature and mature ovaries, and embryonic and immature testes. Western blot analysis and immunocytological detection using specific polyclonal antibodies against amino- and carboxyl-terminal regions of ZOV3 demonstrate that ZOV3 is a plasma membrane-bound glycoprotein that exists in granulosa cells and islets of cells in the theca externa layer of ovarian follicles. The latter islets coincide with those producing estradiol-17 beta. In male and female embryos, production of ZOV3 is first prominent in medullary and seminiferous codes, respectively, of developing gonads. Then, after hatching, it is shifted to the cortex surrounding the primitive follicles in the ovary or is continued weakly in the primary seminiferous tubules in the testis. Expression of the ZOV3 gene and production of ZOV3 are no longer detectable in the mature testis. ZOV3 is unique among the immunoglobulin superfamily proteins in that it is produced predominantly in gonads. Its possible role in differentiation or maintenance of steroidogenic cells in an ovarian follicle is discussed. PMID- 9404002 TI - The expression of tenascin-C with the AD1 variable repeat in embryonic tissues, cell lines and tumors in various vertebrate species. AB - Tenascin-C is a modular glycoprotein composed of domains of amino acid repeats. All forms of tenascin-C have eight constant fibronectin type III repeats, but additional fibronectin type III repeats can be spliced into a variable domain found between the fifth and sixth constant repeats. Four extra repeats, named A, B, C and D, have been examined previously. Here, we have used in situ hybridization to determine the tissue origins of the novel AD1 and AD2 repeats. In the embryonic-day-10 chicken embryo, transcripts encoding the AD2 repeat are limited to the tips of lung bronchioles and the base of feather buds. In contrast the AD1 hybridization signal was widespread. Quantitative in situ hybridization reveals AD1-containing transcripts represent up to 85% of the total tenascin-C mRNA in some tissues (developing bone), and are undetectable in others (e.g. radial glia). Avian and human tumor cell lines were examined for the expression of the AD1 repeat using the reverse transcriptase polymerase chain reaction (RT PCR). Transcripts encoding six different tenascin-C splice variants incorporating the AD1 repeat were found in the fibrosarcoma cell line, QT6. Many human tumor cells, including malignant melanoma and ductal breast carcinoma, were positive for AD1 tenascin-C expression. In addition, we found evidence of AD1 tenascin-C expression in samples of excised human tumors. Our results show that a novel variant may be a major part of the tenascin-C of the embryonic extracellular matrix, and may also be found in the stroma surrounding some human tumors. PMID- 9404003 TI - Hierarchical expression of desmosomal cadherins during stratified epithelial morphogenesis in the mouse. AB - Desmosomes contain two heterogeneous families of specialized cadherins (desmogleins or Dsgs and desmocollins or Dscs), subtypes of which are known to be expressed in tissue-specific and differentiation-dependent patterns in adult epithelial tissues. To examine the temporal and spatial order in which the individual desmosomal cadherins are expressed during stratified epithelial development we have obtained partial cDNA clones of all six murine desmosomal cadherins and have carried out in situ hybridization analysis on E12.5 to E16.5 mouse embryos. The results indicate that the type 2, type 3 and type 1 desmosomal cadherin messages are not obligatorily expressed as pairs during stratified epithelial morphogenesis. Instead the individual genes appear to be transcribed in hierarchical, overlapping temporal and spatial patterns extending from DSG2 to DSC1. DSG2 was the most uniformly expressed message in all E12.5 epithelia, gradually becoming confined to the basal cell layers during epithelial stratification indicating that its transcription was restricted to undifferentiated cells. In contrast, DSC2 message was expressed variably in early epithelia and was strongly upregulated in the suprabasal cell layers during the stratification of wet-surfaced epithelia. DSC3 message was expressed before that of DSG3 in the dental and lingual epithelium where its spatial distribution matched that of DSG2, but after DSG3 in the non-glandular gastric epithelium. DSC3 transcripts became confined to the lower layers of stratifying epithelia but were usually less basally restricted than those of DSG2. Like DSC2, DSG3 mRNA was strongly upregulated in the suprabasal layers of wet-surfaced epithelia as they stratified. Upregulation of DSG1 message was temporally linked to that of DSG3 in all tissues apart from the non-glandular gastric epithelium. PMID- 9404004 TI - MAL mRNA is induced during the differentiation of human embryonal carcinoma cells into neurons and is also localised within specific regions of the human brain. AB - We have found that the MAL gene, which encodes a membrane proteolipid expressed during the late stages of T-lymphocyte maturation, is also activated during neuronal differentiation of NTERA2 human embryonal carcinoma cells following induction with retinoic acid. An RT-PCR fragment with a sequence identical to MAL was found amongst 30 cloned DNA fragments corresponding to genes putatively activated during NTERA2 differentiation and isolated using a differential display PCR screen. PCR and Northern blot analysis with a cloned MAL cDNA as a probe confirmed that MAL is not expressed by undifferentiated NTERA2 EC cells, but is expressed, predominantly as a 1.1 kb transcript, within 7 days of retinoic acid induced differentiation and later in the post-mitotic neurons arising in such cultures. MAL was not expressed in the non-neuronal lineages induced by treatment of NTERA2 cells with the gratuitous inducer hexamethylene bisacetamide. Analysis of cDNA libraries constructed from EC cells, purified neurons and a sub population of non-neuronal cells (ME311+), confirmed that expression of the MAL gene is activated in the neural lineage of NTERA2 differentiation. Using in situ hybridisation we found that MAL is expressed in the human CNS and especially in grey matter of the cerebral cortex, with less in the cerebellum and the amygdala and little or none in subcortical white matter. In contrast to reports concerning the expression pattern of a rat MAL homologue, MAL was expressed in the human brain predominantly in cell bodies which include neurons, correlating with in vitro data from the NTERA2 line. PMID- 9404005 TI - Singular value decomposition of 3-D DNA melting curves reveals complexity in the melting process. AB - The thermal denaturation of synthetic deoxypolynucleotides of defined sequence was studied by a three dimensional melting technique in which complete UV absorbance spectra were recorded as a function of temperature. The results of such an experiment defined a surface bounded by absorbance, wavelength, and temperature. A matrix of the experimental data was built, and analyzed by the method of singular value decomposition (SVD). SVD provides a rigorous, model-free analytical tool for evaluating the number of significant spectral species required to account for the changes in UV absorbance accompanying the duplex--to- single strand transition. For all of the polynucleotides studied (Poly dA-Poly dT; [Poly (dAdT)]2; Poly dG-Poly dC; [Poly(dGdC)]2), SVD indicated the existence of at least 4-5 significant spectral species. The DNA melting transition for even these simple repeating sequences cannot, therefore, be a simple two-state process. The basis spectra obtained by SVD analysis were found to be unique for each polynucleotide studied. Differential scanning calorimetry was used to obtain model free estimates for the enthalpy of melting for the polynucleotides studied, with results in good agreement with previously published values. PMID- 9404006 TI - Temperature and pressure dependence of Shaker K+ channel N- and C-type inactivation. AB - Shaker B potassium channels undergo rapid N-type and slow C-type inactivation. While N-type inactivation is supposed to be mediated by occlusion of the pore by the N-terminal protein structure, the molecular mechanisms leading to C-type inactivation are less well understood. Considering N-type inactivation as a model for a protein conformational transition, we investigated inactivation of heterologously expressed Shaker B potassium channels and mutants thereof, showing various degrees of C-type inactivation, under high hydrostatic (oil) pressure. In addition to the derived apparent activation and reaction volumes (delta V), experiments at various temperatures yielded estimates for enthalpic (delta H) and entropic (T delta S) contributions. N-type inactivation was accelerated by increasing temperature and slowed by high hydrostatic pressure yielding at equilibrium delta H = 76 kJ/mole, T delta S = 82 kJ/mole, and delta V = 0.18 nm3 indicating that the transition to the N-type inactivated state is accompanied by an increase in volume and a decrease in order. N-terminally deleted Sh delta 6-46 constructs with additional mutations at either position 449 or 463 were used to investigate C-type inactivation. In particular at high temperatures, inactivation occurred in two phases indicating more than one process. At equilibrium the following values were estimated for the major inactivation component of mutant Sh delta 6-46 T449A: delta H = -64 kJ/mole, T delta S = -60 kJ/mole, and delta V = 0.25 nm3, indicating that the C-type inactivated state occupies a smaller volume and is more ordered than the noninactivated state. Thus, hydrostatic pressure affects N- and C-type inactivation in opposite ways. PMID- 9404007 TI - The charge state of an ion channel controls neutral polymer entry into its pore. AB - Electrostatic potentials created by fixed or induced charges regulate many cellular phenomena including the rate of ion transport through proteinaceous ion channels. Nanometer-scale pores of these channels also play a critical role in the transport of charged and neutral macromolecules. We demonstrate here that, surprisingly, changing the charge state of a channel markedly alters the ability of nonelectrolyte polymers to enter the channel's pore. Specifically, we show that the partitioning of differently-sized linear nonelectrolyte polymers of ethylene glycol into the Staphylococcus aureus alpha-hemolysin channel is altered by the solution pH. Protonating some of the channel side chains decreases the characteristic polymer size (molecular weight) that can enter the pore by approximately 25% but increases the ionic current by approximately 15%. Thus, the "steric" and "electric" size of the channel change in opposite directions. The results suggest that effects due to polymer and channel hydration are crucial for polymer transport through such pores. PMID- 9404009 TI - Purification and partial characterization of vitellin from the black-legged tick, Ixodes scapularis. AB - Vitellin from the black-legged tick, Ixodes scapularis, was purified from eggs using gel filtration and ion exchange chromatography. The purified protein had a native molecular mass of 480 kDa. Under reducing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis; SDS-PAGE), vitellin was composed of seven polypeptides each at 154, 135, 87, 78, 67, 64 and 35 kDa. The isoelectric point was pH 6.9 and absorption maxima for the yolk protein were 280 and 400 nm. As in other ticks, vitellin from I. scapularis is also a hemoglycolipoprotein. Carbohydrates detected in vitellin were predominantly mannose with a small amount of N-acetylglucosamine. Lipids detected by thin layer chromatography (TLC) were triglycerides, free fatty acids, and cholesterol. Phospholipids associated with vitellin were phosphatidylethanolamine and phosphatidylcholine. Polyclonal serum produced in rabbits recognized vitellin from the eggs and ovaries, and vitellogenin from the hemolymph and fat body in reproductive females. This is the first report on the characterization of yolk proteins from a prostriate tick. PMID- 9404008 TI - Differentially regulated inhibitor-sensitive and insensitive protease genes from the phytophagous insect pest, Helicoverpa armigera, are members of complex multigene families. AB - Ingestion of soybean Kunitz trypsin inhibitor (SKTI) by larvae of the phytophagous insect pest Helicoverpa armigera induced production of inhibitor insensitive protease activity. The induced activity was not due to proteolytic enzymes of different mechanistic classes, but rather to variants of the existing enzymes. Characterization of cDNAs showed that sequences encoding proteins of the serine protease family were abundant in gut tissue of both control and SKTI-fed insects. The majority of serine protease family cDNAs encode enzymes closely homologous to trypsin and chymotrypsin; comparison of these sequences shows variation in amino acid residues within the region which would be in contact with a protein protease inhibitor. More diverged sequences which may not encode active proteases are also present. All the cDNAs examined were found to derive from multigene families; at least 28 different genes are present in the serine protease family. Chronic ingestion of SKTI results in some serine protease encoding mRNA species increasing in level, whereas others decrease. Chymotrypsin encoding mRNAs tend to increase in level as a result of SKTI ingestion, but no clear trend is shown by trypsin-encoding mRNAs. It is suggested that multiple, varying protease-encoding genes are an adaptive mechanism for reducing the deleterious effects of plant protease inhibitors. PMID- 9404010 TI - Occurrence of a common binding site in Mamestra brassicae, Phthorimaea operculella, and Spodoptera exigua for the insecticidal crystal proteins CryIA from Bacillus thuringiensis. AB - Specific binding to midgut membrane proteins is required for the toxicity of insecticidal crystal proteins (ICP) from Bacillus thuringiensis. A direct relationship between toxicity and binding has been proposed. It has been hypothesized that sharing of a single receptor by more than one ICP could lead to the occurrence of multiple resistance in the event of an alteration in the common receptor. Binding of CryIA(a), CryIA(b) and CryIA(c), three structurally related ICPs, has been studied in Phthorimaea operculella, Mamestra brassicae and, Spodoptera exigua using brush border membrane vesicles (BBMV) from the midgut tissue. Using iodinated CryIA(b), the three insects showed similar results: one binding site for CryIA(b), which is shared with CryIA(a) and CryIA(c). The binding site concentrations obtained for CryIA(b) in P. operculella, M. brassicae and S. exigua were 5.1, 16.3 and 2.2 pmol/mg vesicle protein, respectively. In the same way, dissociation constants were 3.8, 5.3 and 0.7 nM. Data show that binding for an ICP does not directly imply toxicity. The occurrence of a common receptor for the CryIA subgroup of ICPs in P. operculella, M. brassicae and S. exigua might theoretically discourage the use of combinations of these ICPs in integrated pest management programmes. PMID- 9404011 TI - The soluble alpha-mannosidases of Drosophila melanogaster. AB - The alpha-mannosidases are implicated in both the catabolism of carbohydrates and the N-linked glycosylation pathway in insects, but little is known of the biochemistry of these glycosidases. In order to study the soluble alpha mannosidases of Drosophila melanogaster we have used artificial fluorogenic substrates for detection of activity in situ following non-denaturing gel electrophoresis. This approach also permitted examination of the mannosidases present in different tissues and the sensitivity of the enzymes to known mannosidase inhibitors. Fluorogenic substrates were also used to determine the pH optima of partly purified mannosidases. We report that D. melanogaster contains several soluble alpha-mannosidase activities. Acidic mannosidases were detected in the gut, fat body and haemolymph of third-instar larvae. The major activity detected in larval guts was a neutral mannosidase presumed to be involved in digestion. PMID- 9404012 TI - Evidence that free fatty acids in trophocytes of Periplaneta americana fat body may be regulated by the activity of phospholipase A2 and cyclooxygenase. AB - Previous studies have shown that palmitic, stearic, oleic and linoleic acid levels in trophocytes prepared from the fat body of male Periplaneta americana are increased following treatment of the cells with hypertrehalosemic hormone (HTH). Melittin, an activator of phospholipase A2, mimicked the action of HTH by increasing the free fatty acid content in a concentration-dependent manner. The increase caused by HTH could be eliminated by pretreatment of the trophocytes with 1 mM 4'-bromophenacyl bromide (BPB), an inhibitor of phospholipase A2. BPB also decreases the concentration of free fatty acids in trophocytes not treated with HTH but by a smaller margin. Nordihydroguaiaretic acid (NDGA) and indomethacin, inhibitors of lipoxygenase and cyclooxygenase, respectively, eliminated the increase in free fatty acids evoked by HTH. In the absence of HTH both inhibitors increased the free fatty acid content of the trophocytes, an effect consistent with the known mode of action of these agents. None of the inhibitors tested, all of which blocked HTH activated trehalose synthesis, prevented activation of phosphorylase by HTH. This is taken as evidence that other downstream sites are also important in the regulation of trehalose production by the fat body. It is suggested that the increase in free fatty acids evoked by HTH, or metabolites of those fatty acids, may regulate the synthesis and release of trehalose from the trophocytes because of potential effects on trehalose phosphate synthase, trehalose 6-phosphate phosphatase, and the trehalose transport mechanism in the trophocyte membrane. PMID- 9404013 TI - Molecular cloning of cDNAs for two pro-phenol oxidase subunits from the malaria vector, Anopheles gambiae. AB - Phenol oxidase exists in insect hemolymph as a zymogen, pro-phenol oxidase (pro PO), which is activated by specific proteolysis in response to infection or wounding. Phenol oxidase catalyses the synthesis of quinones that polymerize to form melanin deposits, which encapsulate parasites and help to seal wounds. Antibodies to pro-PO from Manduca sexta bound to 76, 72, and 71 kDa polypeptide bands from hemolymph of Anopheles gambiae larvae. This antiserum was used to screen a cDNA library from A. gambiae fourth-instar larvae. Full-length clones were isolated for two different pro-POs, designated A. gambiae proPO-p1 and proPO p2, which are 67% identical in nucleotide sequence and 66% identical in deduced amino acid sequence. The A. gambiae pro-PO sequences are more similar to pro-PO from Drosophila melanogaster than to lepidopteran or crustacean pro-PO sequences in the GenBank database. Like the other arthropod pro-POs, the A. gambiae pro-PO sequences lack a signal peptide and have two conserved regions predicted to bind two copper atoms in the active site of the enzyme. The availability of these pro PO cDNAs should be useful in examining the biochemical differences between A. gambiae strains that are refractory or susceptible to Plasmodium infection, and differ in their ability to encapsulate the parasites. PMID- 9404014 TI - Skeletal muscle histology and biochemistry of an elite sprinter, the African cheetah. AB - To establish a skeletal muscle profile for elite sprinters, we obtained muscle biopsy samples from the vastus lateralis, gastrocnemius and soleus of African cheetahs (Acinonyx jubatus). Muscle ultrastructure was characterized by the fiber type composition and mitochondrial volume density of each sample. Maximum enzyme activity, myoglobin content and mixed fiber metabolite content were used to assess the major biochemical pathways. The results demonstrate a preponderance of fast-twitch fibers in the locomotor muscles of cheetahs; 83% of the total number of fibers examined in the vastus lateralis and nearly 61% of the gastrocnemius were comprised of fast-twitch fibers. The total mitochondrial volume density of the limb muscles ranged from 2.0 to 3.9% for two wild cheetahs. Enzyme activities reflected the sprinting capability of the cheetah. Maximum activities for pyruvate kinase and lactate dehydrogenase in the vastus lateralis were 1519.00 +/ 203.60 and 1929.25 +/- 482.35 mumol min-1.g wet wt-1, respectively, and indicated a high capacity for glycolysis. This study demonstrates that the locomotor muscles of cheetahs are poised for anaerobically based exercise. Fiber type composition, mitochondrial content and glycolytic enzyme capacities in the locomotor muscles of these sprinting cats are at the extreme range of values for other sprinters bred or trained for this activity including greyhounds, thoroughbred horses and elite human athletes. PMID- 9404015 TI - Expression of titin isoforms in red and white muscle fibres of carp (Cyprinus carpio L.) exposed to different sarcomere strains during swimming. AB - Titin (also known as connectin) is a striated-muscle-specific protein that spans the distance between the Z- and M-lines of the sarcomere. The elastic segment of the titin molecule in the I-band is thought to be responsible for developing passive tension and for maintaining the central position of thick filaments in contracting sarcomeres. Different muscle types express isoforms of titin that differ in their molecular mass. To help to elucidate the relation between the occurrence of titin isoforms and the functional properties of different fibre types, we investigated the presence of different titin isoforms in red and white fibres of the axial muscles of carp. Gel electrophoresis of single fibres revealed that the molecular mass of titin was larger in red than in white fibres. Fibres from anterior and posterior axial muscles were also compared. For both white and red fibres the molecular mass of titin in posterior muscle fibres was larger than in anterior muscle fibres. Thus, the same fibre type can express different titin isoforms depending on its location along the body axis. The contribution of titin to passive tension and stiffness of red anterior and posterior fibres was also determined. Single fibres were skinned and the sarcomere length dependencies of passive tension and passive stiffness were determined. Measurements were made before and after extracting thin and thick filaments using relaxing solutions with 0.6 mol.l-1 KCl and 1 mol.l-1 KI. Tension and stiffness measured before extraction were assumed to result from both titin and intermediate filaments, and tension after extraction from only intermediate filaments. Compared to mammalian skeletal muscle, intermediate filaments developed high levels of tension and stiffness in both posterior and anterior fibres. The passive tension-sarcomere length curve of titin increased more steeply in red anterior fibres than in red posterior fibres and the curve reached a plateau at a shorter sarcomere length. Thus, the smaller titin isoform of anterior fibres results in more passive tension and stiffness for a given sarcomere strain. During continuous swimming, red fibres are exposed to larger changes in sarcomere strain than white fibres, and posterior fibres to larger changes in strain than anterior fibres. We propose that sarcomere strain is one of the functional parameters that modulates the expression of different titin isoforms in axial muscle fibres of carp. PMID- 9404016 TI - Ontogeny of cardiac rate regulation and brown fat thermogenesis in golden hamsters (Mesocricetus auratus). AB - It was shown previously in infant rats (Rattus norvegicus) that the ability to produce heat in the cold using brown adipose tissue (BAT) is closely related to the ability to maintain cardiac rate. When the limits of BAT thermogenesis were exceeded, interscapular temperature (which reflects the temperature of the interscapular BAT depot) and cardiac rate fell together. As an extension of this earlier study, the relation between BAT thermogenesis and cardiac rate was examined here in the golden hamster (Mesocricetus auratus), a species whose young do not exhibit BAT thermogenesis until the end of the 2nd week postpartum. It was found that 3 to 12-day-old hamsters were unable to increase shivering or nonshivering thermogenesis in the cold and exhibited decreases in cardiac rate that proceeded in lock-step with decreases in interscapular temperature. In contrast, as the thermogenic capability of hamsters increased after 12 days of age, cardiac rate was maintained within narrow limits across a wide range of air temperatures. These results support the hypothesis that heat produced by BAT helps to warm the heart and thus aids in the maintenance of cardiac rate during cold exposure. PMID- 9404017 TI - Cellular signalling of PCH-induced pigment aggregation in the crustacean Macrobrachium potiuna erythrophores. AB - The cellular system responsible for the transduction of the pigment-concentrating hormone (PCH) signal was investigated in erythrophores of the freshwater shrimp, Macrobrachium potiuna. Dose-response curves to the hormone were determined in the absence and in the presence of several drugs that affect sequential steps of the Ca(2+)-dependent signalling pathway. Additionally, the ability of forskolin to induce pigment dispersion was evaluated. Neomycin sulphate (10(-4) and 10(-3) mol.l-1), trifluoperazine (10(-5) and 10(-4) mol.l-1), 1-(5 isoquinolinesulfonlyl)-2-methylpiperazine (10(-7) and 10(-5) mol.l-1) and okadaic acid (10(-7) mol.l-1) significantly (P < 0.05) decreased the responses to PCH. However, okadaic acid at low concentration (10(-9) mol.l-1) and cyclosporin A (10(-6) and 10(-5) mol.l-1) did not significantly (P > 0.05) affect PCH activity. Forskolin (10(-4) mol.l-1) was able to half-maximally reverse the hormone-induced aggregation. Our results suggest that the pigment-concentrating hormone induces pigment aggregation through a Ca(2+)-dependent pathway with a posteriori phosphatase activation, probably the serine/threonine phosphatase 1. PMID- 9404018 TI - Differential effects of glycolytic and oxidative metabolism blockers on the Na-K pump in erythrocytes of the frog, Rana temporaria. AB - This study was undertaken to evaluate the effects of various metabolic blockers on the Na-K-pump activity and ATP content of frog erythrocytes. To eliminate K-Cl cotransport, the frog erythrocytes were incubated in nitrate media at 20 degrees C. Incubation of the red cells in a glucose-free medium for 2 h had no effect on cell ATP content and K+ influx measured as 86Rb uptake for 60 min. The Na(+)-K(+) pump activity was also unchanged in the frog erythrocytes incubated in a glucose free medium containing 10 mM 2-deoxy-D-glucose or adenosine. Unexpectedly, the treatment of red cells with 1-2 mM glycolytic blocker iodoacetate produced a 2 fold increase in the ouabain-sensitive K+ influx. The cell ATP content declined by 9.4% after 2 h of cell incubation with iodoacetate. Incubation of the red cells for 90 min in the presence of 2 mM cyanide, 0.01 mM antimycin A or 5 mM azide resulted in a significant reduction in K+ influx by about 50%, 45% and 32%, respectively. The cell ATP content diminished over 60 min and 120 min of cell incubation with 2 mM cyanide by 15.6% and 31.7% of control levels, respectively. In time-course experiments, a 50% reduction in the K+ influx was observed when the frog erythrocytes were incubated for only 30 min in the presence of 2 mM cyanide. In contrast, 0.01-0.10 mM rotenone, a site I inhibitor, and 0.01 mM carbonyl cyanide m-chlorophenylhydrazone, an uncoupler of oxidative phosphorylation were without effect on K+ influx into frog erythrocytes. These results indicate that about one-half of the Na(+)-K(+)-pump activity in frog erythrocytes is tightly functionally coupled to cytochromes via a separate "membrane-associated" ATP pool. PMID- 9404019 TI - Antigenic relationships among pathogenic Beauveria bassiana with Engyodontium album (= B. alba) and non-pathogenic species of the genus Beauveria. AB - Exoantigenic extracts of 15 isolates belonging to hyalohyphomycosis-causing Beauveria bassiana (1), and Engyodontium album (1), as well as other species of the genus Beauveria (one isolate each of B. brogniartii, B. densa, B. stephanoderis, B. velata, B. vermiconia and six isolates of unknown Beauveria species) were studied. Aqueous-merthiolated extracts derived from 10-day-old Sabouraud's dextrose agar slant cultures (25 degrees C) were concentrated (25X), and reacted against rabbit anti-B. bassiana serum in the presence of partially purified homologous antigen (20X) prepared from 5-week-old shaken cultures (30 degrees C), using a microimmunodiffusion procedure. Beauveria bassiana reference antigen and antiserum reacted to produce four bands of identity. With the exception of E. album, which was negative, extracts of the isolates of B. brogniartii, B. densa, B. stephanoderis, B. velata, B. vermiconia and the unknown Beauveria species all produced 2-4 lines of identity against the homologous anti B. bassiana serum. These results suggested that all the species of the genus Beauveria tested were antigenically related to B. bassiana. Engyodontium album demonstrated antigenic distinctness, however, from B. bassiana and thus supported the validity of this taxon. PMID- 9404020 TI - Quantification of thigmotropism (contact sensing) of Candida albicans and Candida tropicalis. AB - To quantify the thigmotropism, we adapted the our previous method using a chemotaxifilter system in combination with a bioluminescent adenosine triphosphate (ATP) assay based on firefly luciferase-luciferin system and analyzed the relationship between the ability of germ tube formation and thigmotropism of C. albicans and C. tropicalis. Both the ability to form germ tube and the amount of hyphae exhibiting thigmotropism varied depending upon both the species and strains of Candida. C. albicans formed more germ tubes than C. tropicalis. A good correlation was observed between the ability to form a germ tube and the capacity for thigmotropism, and the results gave a level of significance (p < 0.05). Further, SEM observation revealed that relatively long hyphae of C. tropicalis with penetrated through the pores of filter membrane. This phenomenon may be of importance in the development of pathogenesis of C. tropicalis as well as C. albicans. PMID- 9404021 TI - Protein A induced protection against experimental candidiasis in mice. AB - Staphylococcus aureus protein-A (SpA), an important multipotent immunostimulator, increased the resistance to infection with Candida albicans in Swiss albino mice. The mice treated with repeated immunologically active doses (1 microgram/mouse) of SpA, pre- and post-infection, showed one hundred percent protection against experimental candidiasis, and constituted the first such report. Protection could be demonstrated on the basis of leukocyte counts, colony forming unit (CPU) kinetics in liver, spleen, kidney and peritoneal cavity, and phagocytosis by monocyte-derived macrophages. PMID- 9404022 TI - Effect of iron on fluconazole activity against Candida albicans in presence of human serum or monocyte-derived macrophages. AB - Human serum, transferrin, and apotransferrin are known to profoundly inhibit the growth of Candida albicans by iron deprivation. On the other hand, iron overload (iron saturated transferrin) is a serious risk factor for candidiasis in newborn and in leukemic patients. We tested the efficacy of fluconazole and the previously demonstrated synergy of fluconazole and effector cells against C. albicans under iron overload conditions where efficacy might be diminished. We confirm that exogenous iron completely reversed the inhibitory effect of human serum and report that the efficacy of fluconazole against C. albicans was not significantly compromised in a 24 h assay system. Although exogenous iron inhibited fungistatic activity of monocyte-derived macrophages, it did not interfere with the synergistic candidacidal activity of fluconazole and monocyte derived macrophages. In 72 h assays, where fluconazole had candidacidal activity, exogenous iron did not compromise efficacy of fluconazole, and fluconazole activity was often increased. These in vitro results suggest that effectiveness of fluconazole therapy would not be compromised in iron overload situations in vivo. PMID- 9404023 TI - Antigenic similarities to Paracoccidioides brasiliensis in thermo-dependent dimorphic fungi isolated from soil in Botucatu, SP, Brazil. AB - We compared the antigenic characteristics of two thermo-dependent dimorphic fungi isolated from soil in Botucatu, an endemic area of paracoccidioidomycosis (PCM) and Paracoccidioides brasiliensis. The soil isolates grew as cerebriform colonies at 37 degrees C (yeast form) and as cottonous colonies at 25 degrees C (mycelial form). No pathogenicity for ddY mice or hamsters were observed. In immunodiffusion test, there were precipitation bands between the 2 soil isolates and pooled PCM patient sera. There were also common precipitation bands at 21, 50 and 58 kDa between the soil isolates antigens and PCM patient sera by Western blotting, but no gp43 kDa band. No gene for gp 43 kDa protein was detected in the soil isolates by PCR. The fact that these isolates were obtained from an endemic area of PCM and there were some antigenic similarities between the soil isolates and P. brasiliensis in immunodiffusion test and Western-blotting may have some importance in epidemiological surveys done with paracoccidioidin as well interfering with the immune response of the exposed population. PMID- 9404024 TI - Motion edges and regions guide image segmentation by colour. AB - Image segmentation is an important early stage in visual processing in which the visual system groups together parts of the image that belong together, prior to or in conjunction with object recognition. Two principal processes may be involved in image segmentation: an edge-based process that uses feature contrasts to mark boundaries of coherent regions, and a region-based process that groups similar features over a larger scale. Earlier, we have shown that motion and colour interact strongly in image segmentation by the human visual system. Here we explore the nature of this interaction in terms of edge- and region-based processes. We measure performance on a region-based colour segmentation task in the presence of distinct types of motion information, in the form of edges and regions which in themselves do not reveal the location of the colour target. The results show that both motion edges and regions may guide the integrative process required for this colour segmentation task. Motion edges appear to act by delimiting areas over which to integrate colour information, whereas motion similarities define primitive surfaces within which colour grouping and segmentation processes are deployed. PMID- 9404025 TI - Ecological constraints drive social evolution in the African mole-rats. AB - The African mole-rats (family Bathyergidae) are subterranean hystricomorph rodents occurring in a variety of habitats and displaying levels of sociality which range from solitary to eusocial, making them a unique mammalian taxonomic group to test ecological influences on sociality. Here, we use an extensive DNA based phylogeny and comparative analysis to investigate the relationship between ecology, sociality and evolution within the family. Mitochondrial cytochrome-b and 12s rRNA trees reveal that the solitary species are monophyletic when compared to the social species. The naked mole-rat (Heterocephalus glaber) is ancestral and divergent from the Damaraland mole-rat (Cryptomys damarensis), supporting previous findings that have suggested the multiple evolution of eusociality within the family. The Cryptomys genus is species-rich and contains taxa exhibiting different levels of sociality, which can be divided into two distinct clades. A total of seven independent comparisons were generated within the phylogeny, and three ecological variables were significantly correlated with social group size: geophyte density (p < 0.05), mean months per year of rainfall greater than 25 mm (p < 0.001), and the coefficient of rainfall variation (p = 0.001). These results support the food-aridity hypothesis for the evolution of highly social cooperative behaviour in the Bathyergidae, and are consistent with the current theoretical framework for skew theory. PMID- 9404026 TI - The transmission dynamics of antibiotic-resistant bacteria: the relationship between resistance in commensal organisms and antibiotic consumption. AB - We propose a mathematical model of the transmission dynamics of colonization by commensal bacteria within a human community subject to varying levels of antibiotic use designed to control morbidity induced by pathogenic strains of the normally commensal organisms. Colonization is assumed not to induce morbidity in the majority of cases, and antibiotic use is assumed to be related to the arrival and growth of pathogenic strains that give rise to infections including clinical symptoms of disease. In the absence of antibiotic resistance, the model shows how the pattern of antibiotic prescription and use can eliminate the non-pathogenic commensal strains from the host community if the fraction of people taking antibiotics with a defined efficacy exceeds some critical level. The model is extended to take account of the evolution of antibiotic resistance in the commensal population. We assume resistance may be either plasmid-mediated or conferred by selection of low-level pre-existing mutants, and that resistant organisms may experience reduced reproductive fitness. Invasion of the host community by drug-resistant commensals is possible if certain antibiotic prescribing patterns pertain. We calculate these conditions in terms of the transmission parameter of the organism and the level of antibiotic prescription and use. The model is employed to address the issues of how best to use antibiotics in populations harbouring resistant organisms, and when resistant bacteria will out-compete sensitive strains. PMID- 9404027 TI - On the critical behaviour of simple epidemics. AB - We show how ideas and models which were originally introduced to gain an understanding of critical phenomena can be used to interpret the dynamics of epidemics of communicable disease in real populations. Specifically, we present an analysis of the dynamics of disease outbreaks for three common communicable infections from a small isolated island population. The strongly fluctuating nature of the temporal incidence of disease is captured by the model, and comparisons between exponents calculated from the data and from simulations are made. A forest-fire model with sparks is used to classify the observed scaling dynamics of the epidemics and provides a unified picture of the epidemiology which conventional epidemiological analysis is unable to reproduce. This study suggests that power-law scaling can emerge in natural systems when they are driven on widely separated time-scales, in accordance with recent analytic renormalization group calculations. PMID- 9404028 TI - Analysis of dam-calf pairs of BSE cases: confirmation of a maternal risk enhancement. AB - We investigate whether a calf born to a dam that develops bovine spongiform encephalopathy (BSE) (prior or subsequent to the birth) is itself at an enhanced risk of developing BSE. Analyses utilize the main database on reported BSE cases in the British cattle herd maintained by the Central Veterinary Laboratory in Weybridge to trace the dams of BSE-affected animals born following the ruminant feed ban in July 1988. The data reveal a significantly enhanced risk of disease in calves born to BSE-affected dams, with the risk being greatest when birth occurs after the onset of clinical signs of disease in the dam. The dependence of the maternally enhanced risk on the maternal incubation stage at birth argues for a significant component of direct maternal transmission of the aetiological agent of BSE, and offers little support for the hypothesis of genetic predisposition. Using a statistical likelihood model, we obtain estimates of the rate of direct maternal transmission by maternal incubation stage; however, biases in the available data make these values minimum estimates. PMID- 9404029 TI - Parental antagonism, relatedness asymmetries, and genomic imprinting. AB - The theory of inclusive fitness can be modified to consider separate coefficients of relatedness for an individual's maternal and paternal alleles. A gene is said to have parentally antagonistic effects if it has an inclusive fitness benefit when maternally derived, but an inclusive fitness cost when paternally derived (or vice versa). Parental antagonism favours the evolution of alleles that are expressed only when maternally derived or only when paternally derived (genomic imprinting). PMID- 9404030 TI - Infanticide risk and the evolution of male-female association in primates. AB - Year-round association between adult males and females is common in primates, even though internal gestation and lactation predispose males to mate-desertion in the majority of mammals. Because there is little a priori support for alternative explanations, we hypothesized that permanent male-female association in primates serves to reduce the risk of infanticide by strange males whenever females and infants are closely associated. For a phylogenetic test of this hypothesis, we reconstructed the evolution of male-female and female-infant association among primates. The results of Maddison's concentrated changes test confirmed the prediction that mother-infant association, as opposed to infant parking, and female-male association did not evolve independently. Changes in litter size and activity, in contrast, were not significantly associated with evolutionary changes in male-female association. Thus, we demonstrate a fundamental link between primate life history and social behaviour, explain the most basic type of variation in primate social organization, and propose an additional determinant of social organization that may also operate in other mammals. PMID- 9404031 TI - Can skull morphology be used to predict ecological relationships between bat species? A test using two cryptic species of pipistrelle. AB - Can ecological relationships between bat species be predicted largely on the basis of morphology? This question was addressed by investigating skull morphology of two cryptic species of the pipistrelle bat. Since 45 Pipistrellus pipistrellus apparently eats larger prey than 55 P. pipistrellus, we predicted that it would have a larger overall skull size, a larger dentary apparatus, and a larger gape. To test these predictions, variables were measured from skulls of the two cryptic species, and comparisons made between them. In accordance with our predictions, overall skull size was larger in 45 P. pipistrellus than in 55 P. pipistrellus, and 45 P. pipistrellus had a longer lower jaw and the distance between the jaws at maximum gape was larger. In addition, 45 P. pipistrellus had longer upper canines, which may allow it to pierce harder prey items than 55 P. pipistrellus. Only some aspects of dietary differences between the two cryptic species could be explained by differences in skull morphology, and we suggest that empirical data, at least on diet and habitat use, are also required to explain mechanisms of resource partitioning among species in bat communities. PMID- 9404033 TI - Accurate peptide sequencing by post-source decay matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - Partial 18O-labeling of peptides has been applied to post-source decay experiments in a matrix-assisted laser desorption/ionization time-of-flight mass spectrometer. The ions which originate from the carboxyl terminus of a peptide partially retain 18O atoms which have readily been incorporated into the C terminal carboxyl groups during enzymatic hydrolysis in a buffer containing 40 atom percent H218O. The isotopic resolution of singly charged precursor ions and their product ions obtained for peptides up to ca. 2800 Da has been achieved using the delayed extraction method, which permits the rapid identification and assignment of the 18O-labeled and non-labeled ion species in the PSD spectra. The results obtained from several 18O-labeled peptides, derived from an enzymatic digest, demonstrate the accuracy and reproducibility of the present method, which will be in widespread use for protein identification via database searching or for investigations of totally unknown proteins. PMID- 9404032 TI - Motile areas of leech neurites are rich in microfilaments and two actin-binding proteins: gelsolin and profilin. AB - Cell motility is produced by changes in the dynamics and organization of actin filaments. The aim of the experiments described here was to test whether growing neurites contain two actin-binding proteins, gelsolin and profilin, that regulate polymerization of actin and affect non-neuronal cell motility. The distribution of gelsolin, profilin and the microfilaments was compared by immunocytochemistry of leech neurons growing in culture. We observed that microfilaments are enriched in the peripheral motile areas of the neurites. Both gelsolin and profilin are also concentrated in these regions. Gelsolin is abundant in filopodia and is associated with single identifiable microfilament bundles in lamellipodia. Profilin is not prominent in filopodia and shows a diffuse staining pattern in lamellipodia. The colocalization of gelsolin and profilin in motile, microfilament-rich areas supports the hypothesis that they synergistically regulate the actin dynamics that underlie neurite growth. PMID- 9404034 TI - Stable isotope ratio analysis of amino acids: the use of N(O,S)-ethoxycarbonyl trifluoroethyl ester derivatives and gas chromatography/mass spectrometry. AB - N(O,S)-Ethoxycarbonyl trifluoroethyl amino acid esters are formed by the reaction of amino acids with ethylchloroformate plus trifluoroethanol plus pyridine. The use of these derivatives for a rapid and sensitive determination of specific enrichment of stable isotopically labeled tracer amino acids in blood plasma and protein hydrolysates, by using gas chromatography/electron impact mass spectrometry, is discussed. PMID- 9404035 TI - Characterization of the conformations of antigenic peptides of protein lactate dehydrogenase (LDH-C4) by electrospray ionization mass spectrometry. AB - The conformations of several rationally designed antigenic peptides that mimic, to varying degrees, an antibody-binding region of protein lactate dehydrogenase isozyme (LDH-C4) are investigated by deuterium/hydrogen exchange and electrospray ionization mass spectrometry (ESI-MS). The approach involves monitoring the reverse-exchange of deuterium, incorporated at the labile sites in the peptides, with hydrogen as a function of time by ESI-MS. Idealized forms of a segment of the native antigen are shown to be more conformationally restricted than the native peptide based on level of deuterium that remains incorporated at the labile sites over time. From the number of amide groups of the peptide backbone that retain deuterium, estimates of the helical content of each peptide have been measured that are in close agreement with those determined by Fourier transform infrared (FTIR) spectroscopy in separate experiments. A single amino acid substitution in the idealized helical construct results in a conformational change easily detected by the deuterium exchange ESI-MS method. The approach is shown to be a viable method for characterizing the conformations of protein antigens at the local level and for screening the conformations of antigenic peptides designed to elicit optimal immune responses. PMID- 9404036 TI - Matrix-assisted laser desorption/ionization mass spectrometry of proteins extracted directly from sodium dodecyl sulphate-polyacrylamide gels. AB - A new strategy for the characterization of Coomassie Brilliant Blue stained SDS PAGE separated proteins by UV-MALDI-MS is reported. The proteins are extracted directly from the polyacrylamide gel by treatment with an organic solvent mixture consisting of formic acid, acetonitrile, isopropanol and water in an ultrasonic bath. A fraction of the supernatant is then mixed directly with the matrix solution and measured by MALDI-MS. High quality spectra could be obtained from gels which were loaded with 6 pmol of myoglobin. Compared to other methods based on electroblotting or electroelution this method is much simpler and less time consuming. The sensitivity is higher than or comparable to the Coomassie Blue staining procedure for proteins up to about 25 kDa. Another advantage is that mass shifts due to charging effects of the membranes, which are common if membranes are mounted directly on the sample target, can be avoided. However, all proteins studied showed slightly higher masses than expected which reduces mass accuracy to 0.2-0.3%. This is presumably partly due to formylation of serine or threonine residues during incubation in formic acid. Gel electrophoresis induced modifications can contribute as well. The possibility of further characterizing the remaining part of the supernatant after extraction by means of proteolytic digestion is also demonstrated. The knowledge of both molecular weight of the whole protein and of the proteolytic fragments increases specificity for protein identification by searching in sequence databases. PMID- 9404037 TI - Assessment of bog-body tissue preservation by pyrolysis-gas chromatography/mass spectrometry. AB - Flash pyrolysis-gas chromatography/mass spectrometry (py-GC/MS) was used to assess the quality and mechanism of protein preservation in the tissue of Iron Age bog bodies from Lindow, UK, and south-eastern Drenthe, The Netherlands. Abundant pyrolysis products of the fresh skin tissue, including 2,5 diketopiperazines of Pro-Gly, Pro-Ala, Pro-Val, Pro-Pro and Hyp, were readily assigned to specific amino acid or dipeptide moieties. Comparison of the pyrolysates of the bog-body tissues with that of modern samples revealed qualitative similarities suggesting good preservation of the collagen and non collagenous proteins in the ancient tissues. Examination of the pyrolysates of samples of fresh calf skin, which had been treated with various vegetable tanning agents, clearly revealed markers of non-hydrolysable tannins including 1,2 benzenediol, 1,3-benzenediol and 1,2,3-benzenetriol, although chromatographic quality inevitably diminished with increasing functionalization of the compounds. Such markers were not detected in the pyrolysates of the bog-body tissues. Instead 4-isopropenylphenol, a characteristic pyrolysis product of Sphagnum moss, was detected in both solvent-extracted and base-treated samples of tissue. The presence of 4-isopropenylphenol in the pyrolysates of the bog-body tissues provides evidence that their preservation involves reactions of amino acids with sphagnum acid, and possibly other agents derived from the peat. The study constitutes the first chemical characterization of the pyrolysis products of modern and ancient collagen. PMID- 9404038 TI - High-performance liquid chromatography matrix-assisted laser desorption/ionization time-of-flight mass spectrometry peptide fingerprinting of tarantula venoms in the genus Brachypelma: chemotaxonomic and biochemical applications. AB - Precise identification of arthropod species is fundamental in venom research, particularly in groups where taxonomy problems remain unsolved. High-performance liquid chromatography and matrix-assisted laser desorption/ionization time-of flight mass spectrometry (MALDI-TOFMS) analysis of crude venoms of six tarantula species in the genus Brachypelma showed that the characteristic chromatographic and peptide ion profiles obtained can be used to discriminate amongst closely related species. This method permits rapid mass fingerprinting of large numbers of samples in a reproducible manner, and offers a powerful systematic tool in combination with morphological methods for the classification of tarantula species. The sensitivity and precision of the method may offer a way to solve complex taxonomic relationships not easily resolved by morphological measurements, in a non-destructive manner. Additionally, peptide mapping of crude venoms by MALDI-TOFMS will speed up the discovery of novel ligands of neuronal receptors, since major venom components of related species share a high sequence homology and are likely to possess similar pharmacological properties. PMID- 9404039 TI - Rapid profiling of E. coli proteins up to 500 kDa from whole cell lysates using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry was used to rapidly detect and profile large proteins from Escherichia coli whole cell lysates in the mass range 25-500 kDa. The bacterial samples were treated with guanidine hydrochloride and Triton X-100 to disrupt and solubilize the large inner membrane proteins. A sample preparation involving a nitrocellulose polymer film, and alpha-cyano-4-hydroxycinnamic acid, sinapinic acid or caffeic acid as matrix was utilized to rapidly monitor the presence of induced and repressed protein synthesis in response to L-arabinose catabolism in E. coli cells. The results were compared to those of 1-D or 2-D gel electrophoresis. PMID- 9404040 TI - Rapid determination of corticosteroids in urine by combined solid phase microextraction/liquid chromatography/mass spectrometry. AB - Solid-phase microextraction combined with fast short-column liquid chromatography/mass spectrometry (SPME/LC/MS) was used for isolating and analysing 11 corticosteroids and 2 steroid conjugates from urine samples. Several SPME parameters such as polarity of fibres, extraction time and effect of ionic strength, were investigated, and their impact on the SPME/LC/MS technique was studied. To demonstrate the potential of the SPME/LC/MS method, its application to the determination of steroids in human urine was investigated. The method was shown to be sensitive with detection limits between 4 and 30 ng/mL and precision between 4.9 and 11.1% RSD through the use of an internal standard technique. The versatility of the method was also exhibited by its excellent linearity in the concentration range of 20 to 20,000 ng/mL in urine for all investigated compounds. A comparison of the SPME/LC/MS method with LC/MS methods utilizing traditional sample preparation techniques, shows that the former offers similar performance in terms of precision, linearity and detection limits, but is clearly easier to use and faster to perform. PMID- 9404041 TI - Covalent binding of 2,4,6-trinitrotoluene to human hemoglobin. Evidence for protein adducts probed by electrospray ionization mass spectrometry. PMID- 9404042 TI - Breast cancer, radon, and brassieres??? PMID- 9404043 TI - ATSDR science panel on the bioavailability of mercury in soils: lessons learned. AB - On the basis of discussion and analysis during and following an ATSDR science panel on the bioavailability of mercury in soils, it is apparent that the default assumption of 100% relative bioavailability for mercury-contaminated soils is excessively conservative. However, current knowledge does not allow the development of default assumptions or guidelines for determining relative bioavailability of mercury in soils. Until such default assumptions or guidelines can be developed, site-specific assays of bioavailability, preferably using either animal bioassays or validated in vitro techniques, may provide the best approach for estimating soil-mercury bioavailability. PMID- 9404044 TI - Current views on the oral bioavailability of inorganic mercury in soil: implications for health risk assessments. AB - Due to the presence of mercury at a number of major contaminated sites in the United States, the bioavailability of inorganic mercury in soil following ingestion has emerged as an important public health issue. Studies of the leachability/solubility of inorganic mercury in soil have shown that it is largely immobile, thereby suggesting that it will not be readily available for absorption in the gastrointestinal tract. Ignoring the effect of the soil matrix on decreasing bioavailability may result in a substantial overprediction of risks due to ingestion of contaminated soil. This paper discusses current knowledge about the oral bioavailability of inorganic mercury in soil and offers suggestions about how these data may be applied in human health risk assessment. Though precise estimates are not available, in vivo and in vitro estimates of the bioavailability of different inorganic mercury species in different matrices suggest that the bioavailability of mercury in soil is likely to be significantly less, on the order of at least three- to tenfold, than the bioavailability of mercuric chloride, the species used to derive the toxicity criteria for inorganic mercury. Because bioavailability can vary significantly with soil type, soil aging, the presence of co-contaminants and other factors, it is suggested that whenever the fiscal aspects justify a more precise estimate of bioavailability, site-specific estimates be developed. To develop a database for identifying a less expensive and more efficient method for estimating bioavailability, it is suggested that in vivo studies be conducted concurrently with in vitro studies. However, due to the lack of precision associated with the derivation of the most widely-used health guidance value for inorganic mercury (the USEPA RfD), additional work to address the uncertainties in the RfD is recommended. PMID- 9404045 TI - Evaluation of methods for assessing the oral bioavailability of inorganic mercury in soil. AB - The risks associated with environmental exposures to inorganic mercury are typically assessed based on toxicity studies conducted with the soluble salt, mercuric chloride (HgCl2). Evidence indicates, however, that inorganic mercury is present in soil as a variety of compounds and that oral absorption of inorganic mercury decreases with a decrease in the solubility of the mercury compound being studied. Thus, while HgCl2 is approximately 15-20% bioavailable, the bioavailability of cinnabar (HgS) may be 30- to 60-fold less. The solubility and, hence, bioavailability of inorganic mercury in soil is expected to be substantially less than that of HgCl2 due to the presence of less soluble compounds and their interactions with soil constituents. Quantification of this difference in bioavailability is important in assessing potential risks associated with exposure to mercury-containing soil. A review of available studies supports the expectation that mercury bioavailability in soils will be reduced. This paper reviews methods for assessing soil metal absorption with consideration of the characteristics of the oral absorption of elemental and inorganic mercury that should be evaluated in designing additional studies. Because of the very slow elimination of mercury in some species, it is recommended that a repeated-dose study be conducted. Such a study would yield an estimate of relative bioavailability based on a comparison of tissue mercury concentrations in animals ingesting soil with those of animals receiving HgCl2. The dose, age, gender, and species of animal selected are not expected to affect relative bioavailability estimates; however, it is recommended that studies be conducted in two animal species. Rats should be used because they have been used in many studies of mercury absorption and toxicity. A species of large animals such as monkeys, swine, or dogs should also be used to provide confirmation in a species with greater similarities to humans in gastrointestinal physiology and anatomy. Other critical factors in designing these studies, such as selection and characterization of soil samples, are also addressed. PMID- 9404046 TI - The environmental geochemistry and bioaccessibility of mercury in soils and sediments: a review. AB - There have been many studies of mercury geochemistry in the environment and its bioconcentration/bioaccumulation through the aquatic food chain. However, there is a dearth of information regarding the bioaccessibility of mercury in human receptors exposed primarily by soil ingestion. This paper reviews the current state of knowledge of mercury bioaccessibility and speciation in soils, and the utility of speciation methods to estimate mercury bioaccessibility. We conclude that additional research is necessary to determine: (1) whether analytical measurements can adequately determine the bioaccessibility of mercury in sediments and soils; (2) the accuracy of in vitro analyses in assessing mercury bioaccessibility; (3) the ability of mercury to cross tissue membranes of the mouth, esophagus, stomach, and the small and large intestines; (4) the speciation and distribution of mercury in biological fluids; and (5) mercury bioavailability using an in vivo animal model relevant to human gastrointestinal tract conditions. PMID- 9404048 TI - A physiologically-based pharmacokinetic model assessment of methyl t-butyl ether in groundwater for a bathing and showering determination. AB - Methyl t-butyl ether (MTBE) is a gasoline additive that has appeared in private wells as a result of leaking underground storage tanks. Neurological symptoms (headache, dizziness) have been reported from household use of MTBE-affected water, consistent with animal studies showing acute CNS depression from MTBE exposure. The current research evaluates acute CNS effects during bathing/showering by application of physiologically-based pharmacokinetic (PBPK) techniques to compare internal doses in animal toxicity studies to human exposure scenarios. An additional reference point was the delivered dose associated with the acute Minimum Risk Level (MRL) for MTBE established by the Agency for Toxic Substances and Disease Registry. A PBPK model for MTBE and its principal metabolite, t-butyl alcohol (TBA) was developed and validated against published data in rats and humans. PBPK analysis of animal studies showed that acute CNS toxicity after MTBE exposure can be attributed principally to the parent compound since the metabolite (TBA) internal dose was below that needed for CNS effects. The PBPK model was combined with an exposure model for bathing and showering which integrates inhalation and dermal exposures. This modeling indicated that bathing or showering in water containing MTBE at 1 mg/L would produce brain concentrations approximately 1000-fold below the animal effects level and twofold below brain concentrations associated with the acute MRL. These findings indicate that MTBE water concentrations of 1 mg/L or below are unlikely to trigger acute CNS effects during bathing and showering. However, MTBE's strong odor may be a secondary but deciding factor regarding the suitability of such water for domestic uses. PMID- 9404049 TI - Biosphere model for high level waste repository. AB - Method for environmental impact modeling involving uncertainty, overcoming the disadvantage of providing only one upper bound based on accumulated effects from all extreme events. This method provides a suite of upper and lower bounds based on any subset of such extreme events, to be chosen among by the decision maker. PMID- 9404050 TI - A simulation model for occupational tuberculosis transmission. AB - A simulation model of tuberculosis (TB) transmission among hospital employees is described. A hypothetical cohort of 1000 workers was divided into low-, medium-, and high-risk groups. The number of TB patients admitted daily was treated as a Poisson random variable. A patient imparted a daily infection risk that was identical for all workers within a risk group but that varied between risk groups. In some scenarios, infected employees were assigned a daily risk of developing TB disease. If disease developed, the individual remained on the job for 3 calendar weeks and imparted a substantial infection risk to 25 close contacts. Simulations were run over 5-year intervals. Cumulative infection incidence increased over time and with more TB patients admitted. Given a scenario in which there were 600, 300, and 100 susceptibles in the low-, medium-, and high risk groups, respectively, 50 TB patients admitted annually and accounting for disease among infected employees, at 5 years there were approximately 100 primary infections (due to infection by patients), 40 secondary infections (due to infection by diseased coworkers), five primary disease cases, and two secondary disease cases. The input parameter values and simulation outcomes were reasonably consistent with the sparse information reported in the literature. PMID- 9404051 TI - A unified approach to a class of stochastic carcinogenesis models. AB - Survival and hazard functions play a pivotal role in carcinogenesis modeling. This paper provides a unified approach to computing these two quantities by classifying a large class of carcinogenesis models from a computational point of view, and prescribes specific computational recipes according to this classification. PMID- 9404052 TI - Using GIS in risk analysis: a case study of hazardous waste transport. AB - This paper provides an illustration of how a geographic information system (GIS) can be used in risk analysis. It focuses on liquid hazardous waste transport and utilizes records archived by the London Waste Regulatory Authority. This data source provides information on the origin and destination of each waste stream, but not the route followed during transport. A GIS was therefore employed to predict the paths used, taking into account different routing criteria and characteristics of the available road network. Details were also assembled on population distribution and ground-water vulnerability, thus providing a basis for evaluating the potential consequences of a waste spillage during transport. Four routing scenarios were implemented to identify sections of road which consistently saw heavy traffic. These simulations also highlighted that some interventions could lead to risk tradeoffs rather than hazard mitigation. Many parts of the research would not have been possible without a GIS, and the study demonstrates the considerable potential of such software in environmental risk assessment and management. PMID- 9404053 TI - The vulnerability of the Nevada visitor economy to a repository at Yucca Mountain. AB - This article reviews the studies commissioned by the Nevada Nuclear Waste Project Office to estimate the economic impact of a high-level nuclear waste repository at Yucca Mountain. Case studies found that visitor impacts occur for some analogous facilities, but not for others. Assessments of behavioral intent indicate that at least some economic agents would avoid visiting Nevada under repository scenarios. A third set of studies tested the risk-aversion and negative-imagery models of visitor decision making; people avoid visiting places associated with either a significant health risk or negative imagery, but it has yet to be shown that a repository would induce these perceptions in nearby places. In sum, the NWPO-sponsored studies suggest the potential for visitor impacts, but do confirm that these effects will occur. PMID- 9404054 TI - Variability in PAH-DNA adduct measurements in peripheral mononuclear cells: implications for quantitative cancer risk assessment. AB - Biomarkers such as DNA adducts have significant potential to improve quantitative risk assessment by characterizing individual differences in metabolism of genotoxins and DNA repair and accounting for some of the factors that could affect interindividual variation in cancer risk. Inherent uncertainty in laboratory measurements and within-person variability of DNA adduct levels over time are putatively unrelated to cancer risk and should be subtracted from observed variation to better estimate interindividual variability of response to carcinogen exposure. A total of 41 volunteers, both smokers and nonsmokers, were asked to provide a peripheral blood sample every 3 weeks for several months in order to specifically assess intraindividual variability of polycyclic aromatic hydrocarbon (PAH)-DNA adduct levels. The intraindividual variance in PAH-DNA adduct levels, together with measurement uncertainty (laboratory variability and unaccounted for differences in exposure), constituted roughly 30% of the overall variance. An estimated 70% of the total variance was contributed by interindividual variability and is probably representative of the true biologic variability of response to carcinogenic exposure in lymphocytes. The estimated interindividual variability in DNA damage after subtracting intraindividual variability and measurement uncertainty was 24-fold. Inter-individual variance was higher (52-fold) in persons who constitutively lack the Glutathione S Transferase M1 (GSTM1) gene which is important in the detoxification pathway of PAH. Risk assessment models that do not consider the variability of susceptibility to DNA damage following carcinogen exposure may underestimate risks to the general population, especially for those people who are most vulnerable. PMID- 9404056 TI - Genetic analysis of plasmid-specific pheromone signaling encoded by pPD1 in Enterococcus faecalis. AB - Certain plasmids in Enterococcus faecalis encode a mating response to recipient produced peptide sex pheromones. Targeted disruption of tra genes on pPD1 suggested that TraA plays a central role in the plasmid-specific pheromone signaling pathway. TraA functioned as a negative regulator for the pheromone inducible conjugal transfer. Complementation analysis of pPD1 tra gene mutants by pAD1 suggested that the pheromone binding function of TraC was non-specific between these plasmids, but the function of TraA and the pheromone shutdown function of TraB are plasmid-specific. PMID- 9404055 TI - Effects of lecithin addition in oil or water phase on the stability of emulsions made with whey proteins. AB - The effects of lecithin addition in oil or water phase on the stability of oil-in water emulsions made with 0.1 wt% whey protein and 10 wt% n-tetradecane at neutral and acidic pH were studied by monitoring the gravitational creaming and phase separation. The effects of lecithin addition on the interfacial behavior of beta-lactoglobulin were also studied to compare with the results of emulsion stability. At neutral pH, crude phosphatidylcholine (PC) from egg yolk or soybean increased the stability of the emulsion made with protein and lowered the interfacial tension of protein films more effectively than pure egg PC. A more remarkable effect on both the emulsion stability and the interfacial tension was found when crude PC was added in the oil phase rather than in the water phase. The purity of lecithins and the way to add them are suggested to be very important to make a stable emulsion with protein. On acidic pH (4.5 or 3.0), the increased creaming or phase separation in a whey protein-stabilized emulsion, but the lowered interfacial tension of beta-lactoglobulin films, were found upon the addition of pure or crude PC in oil or water phase. These results suggest that in acidic pH, densely packed films may be formed on a planar oil-water interface, but not on adsorbed layers around oil droplets in an emulsion. PMID- 9404057 TI - Contents of resveratrol, piceid, and their isomers in commercially available wines made from grapes cultivated in Japan. AB - The presence of resveratrol (3,5,4'-trihydroxystilbene) and its beta-glucoside, piceid (resveratrol-3-beta-D-glucopyranoside), together with their isomers in wine appears to be one of the beneficial factors conferring a protective effect against cardiovascular disease through red wine ingestion. A total of 42 red and white wines was collected in areas from Hokkaido to Kyushu in Japan. The wines were fractionated with a C18 Sep-pak cartridge, and the active principles were eluted with ethyl acetate. Crude trans- and cis-piceid were extracted from a Chinese medicine, 'Kojohkon' (Polygonum cuspidatum), and their retention times and UV absorption were confirmed by HPLC. trans- and cis-Resveratrol, and trans- and cis-piceid were analyzed in a short C18 HPLC column, and cis-resveratrol was quantified from the amount of cis-isomer converted from authentic trans resveratrol that had been treated by UV irradiation. The content of piceid is shown as the resveratrol equivalent. The average content of total stilbene compounds was 4.37 mg/liter in red wines, while only 0.68 mg/liter in white wines. Red wines made from Pinot noir, Merlot, and Zweigeltrebe grapes all had a high resveratrol content. PMID- 9404058 TI - Specific interaction of cytokinins and their analogs with rotenone-sensitive internal NADH dehydrogenase in potato tuber mitochondria. AB - Effects of cytokinins were studied on rotenone-sensitive NADH dehydrogenase in mitochondria from fresh potato tubers (Solanum tuberosum), in consideration of the operation of external and rotenone-insensitive internal NADH dehydrogenases that has not been fully accounted for in previous studies. In submitochondrial particles (smp), zeatin was only weakly active, and zeatin riboside (ZR) was inactive. Inhibition rates at 400 microns of isopentenyladenine (iP) and isopentenyladenosine (iPA) were 45% and 30%, respectively, and that of BA (BA) was 64%. In intact mitochondria, the inhibition by iP and BA significantly increased, I50 being 50 and 250 microM, respectively, but that by zeatin and iPA decreased. A structure-activity study showed that hydrophobic and steric factors are important for the activity. Cytokinins inhibited the electron flow via natural quinone more strongly than that via synthetic quinone. These results suggest that among the cytokinins the species that can regulate the electron transport is iP rather than its riboside or zeatin. PMID- 9404059 TI - Different responses of apolipoprotein A-I, A-IV, and B gene expression during intestinal adaptation to a massive small bowel resection in rats. AB - Gene expression of apolipoproteins (apo) A-I, A-IV, and B, the predominant protein components of chylomicrons, was investigated in the residual ileum after a massive small bowel resection in rats. A Northern blot analysis showed that the apo A-IV mRNA level, but not the apo A-I and B mRNA levels, in the ileum was significantly higher in the resected rats than in the sham-operated rats 24 h and 2 wk post-surgery. RT-PCR coupled with a primer extension assay revealed that the apo B-48 mRNA/apo B-100 mRNA ratio, i.e., apo B mRNA editing, in the ileum was unchanged by the resection. It is thus concluded that, among the major intestinal apolipoproteins, apo A-IV is the only one whose gene expression is influenced by loss of the proximal intestine. PMID- 9404060 TI - Metabolic behavior of angiotensins in normotensive human plasma in the supine and upright postures. AB - The effect of activating the renin-angiotensin system on the metabolism of angiotensins (ANGs) in normotensive human plasma was investigated. In normotensive supine human plasma, four peptides with in vitro angiotensin I converting enzyme (ACE) inhibitory activity which correspond to the sequence of ANG (3-8), ANG (4-8), ANG (5-8), and ANG (3-4) existed at a concentration of > 39 fmol/ml of plasma. When activating the renin activity by keeping upright in posture for 60 min, ANG II and the four peptides significantly increased as compared with the levels in the supine posture, except for ANG I. In particular, Val-Tyr corresponding to ANG (3-4) in the upright posture was about 4-fold more than the value in the supine posture, and was predominantly present (447 fmol/ml of plasma) as well as ANG I. As a result of in vitro degradation tests on ANGs, ANG (3-4) was produced from ANG I, and not from ANG II, III or (3-8), during the 30-min incubation. PMID- 9404061 TI - Water permeability of plasma membranes of cultured rice, grape, and CH27 cells measured dielectrically. AB - The capacitance of suspensions of cultured rice cells (Oryza sativa L. ssp. japonica), grape cells (Vitis sp.), and CH27 cells originated from murine B-cell lymphoma was measured in the frequency range of 0.2 to 10 MHz. The relationship between the increase in capacitance caused by the presence of cells at 0.4 MHz, delta C, and the cell density was linear. Measurement of capacitance was useful in measurement of transitional changes in cell volume under external osmotic stress when sucrose was added. From the course of volume changes with such stress, the water permeabilities of the plasma membrane, Lp, were measured to be 0.015, 0.020, and 0.090 pm/(s.Pa) at 25 degrees C, for rice cells, grape cells, and CH27 cells, respectively. The smaller Lp for plant cells seemed to explain why preservation of plant cells by freezing is more difficult than for animal cells. From the temperature dependence of Lp, the apparent activation energies were calculated to be 12.0 +/- 2.9 and 13.0 +/- 5.2 kcal/mol for rice cells and CH27 cells, respectively. PMID- 9404062 TI - A model system for convenient fluorescent labeling of sugar chain in taka-amylase A. AB - A convenient detection of sugar chains in Taka-amylase A (TAA) was done by using 40 micrograms of enzyme, where a decrease in the UV absorption of NaIO4 during the periodate oxidation reaction was monitored. The periodate-oxidized sugar chain was labeled with a fluorescent reagent, N-1-ethylenediaminonaphthalene (EDAN), by incubation at pH 9.5 and 30 degrees C for 1 h. The excess EDAN was removed by either quenching with o-phthaladehyde or Bio-Gel P-2 gel adsorption. Among the peptide fragments prepared from the EDAN-labeled TAA, a fluorescent peptide corresponding to the sugar chain was distinguished by the ODS column. These results suggest that periodate oxidation and subsequent fluorescent labeling were useful for the sensitive analysis of various glycoprotein samples. PMID- 9404063 TI - Growth factors for a primary chick muscle cell culture from shochu distillery by products. AB - An unidentified growth factor (UGF) was separated from shochu distillery by products (SDBP) and its effect on the growth of a primary chick muscle cell culture was investigated. Chick muscle cells were isolated from fertile eggs (13 day-old embryos) of commercial broilers. UGF was separated on Sephadex LH-20 with a solvent system of water-methanol-ethylene dichloride (10:90:20, v/v), and the fraction eluted between 136 and 164 min was collected (fraction I). Fraction I was further purified by HPLC with an Inertsil ODS-2 column using a solvent system of methanol-butanol (80:20, v/v). Three fractions having retention times of 3.76, 4.57, and 5.12 min were collected and are referred to as fraction A, B, and C, respectively. In experiment 1, chick muscle cells were cultured in an m-199 medium containing 0.001, 0.01, or 0.1% of fraction I. In experiment 2, chick muscle cells were cultured with 0.01 or 0.005% of each fraction A, B, and C. Creatine kinase (CK) activity, protein and DNA contents were measured as indices of myotube growth, cell growth and cell proliferation, respectively. N tau methylhistidine (N tau-MH) release from the muscle cell was also measured to observe the effect on proteolysis. In experiment 1, the protein content was significantly (p < 0.05) increased by fraction I, despite the low dose level. CK activity was significantly (p < 0.05) higher than the control when 0.001% of fraction I was added to the medium. However, increasing the level beyond 0.01% did not further increase the CK activity. The DNA content was not significantly changed. In experiment 2, the protein content, CK activity, and DNA content were significantly (p < 0.05) higher when fractions A and B were added to the medium. However, this was not the case when fraction C was added. N tau-MH release was significantly (p < 0.05) higher when fraction A was added, but, was significantly (p < 0.05) lower when fraction B was added, while fraction C had no effect on N tau-MH release. The present results show that SDBP contained two growth-promoting factors for a primary chick muscle cell culture, although their modes of action may be different. PMID- 9404064 TI - Cloning and sequence analysis of the gene (alyII) coding for an alginate lyase of Pseudomonas sp. OS-ALG-9. AB - Pseudomonas sp. OS-ALG-9 produces several kinds of alginate-degrading enzymes both intra- and extracellularly. As a second alginate lyase of this bacterium, the gene encoding alyII has been cloned in Escherichia coli JM109 by shotgun techniques and then sequenced. The alyII gene has an open reading frame of 2141 bp encoding 713 amino acid residues with a calculated molecular mass of 79,803 Da. The deduced amino acid sequence did not show any extensive similarity with those of other known alginate lyases, however, hydrophobic cluster analysis showed that alyII belonged to class 3 of alginate lyases. The alginate lyase from E. coli harboring the alyII gene showed a single active band, which coincided with one of four major alginate lyases from the crude cell extracts of Pseudomonas sp. OS-ALG-9 on a zymogram. PMID- 9404065 TI - Secretion of human interleukin-2 in biologically active form by Bacillus brevis directly into culture medium. AB - We constructed an efficient system for synthesis and secretion of human interleukin-2 (IL-2) by Bacillus brevis. The secretion vector we constructed had strong promoters and contained the region coding for the signal peptide of the gene for B. brevis 47 cell-wall protein, followed directly by the gene encoding mature IL-2. Modification of the signal peptide and use of a protease-deficient mutant of B. brevis HPD31 increased productivity. When the signal peptide was more basic near its amino terminal and more hydrophobic in the middle region, IL 2 production increased 20 fold. Production by the mutant harboring the secretion vector was four fold that of the parent harboring the same plasmid. The yield of IL-2 increased further to 0.12 g/liter, when cultural conditions were made optimal, such by the addition of Tween 40 to the medium. The IL-2 produced by B. brevis had the same biological activity as authentic IL-2. Biologically active human IL-2 was produced efficiently and secreted directly into the medium by B. brevis. PMID- 9404067 TI - Hyperproduction of L-threonine by an Escherichia coli mutant with impaired L threonine uptake. AB - An efficient production strain for L-threonine fermentation was derived from Escherichia coli by multiple rounds of mutation programs that aimed at deregulation of the L-threonine biosynthetic pathway and blocking of L-threonine degradation pathways. When the optimum amount of DL-methionine was added, this strain KY10935, an L-methionine auxotroph, gave 100 g/liter L-threonine after 77 h cultivation. In this strain, key enzymes in the L-threonine biosynthetic pathway were highly derepressed, but some were inhibited by lower concentrations of L-threonine than the accumulated level. Such incomplete deregulation of the pathway was accounted for by the intracellular concentration of L-threonine being lower than the extracellular level. In an assessment of L-threonine transport in terms of phenotypic growth responses to the amino acid, L-threonine-auxotrophic mutants with a lesion in the L-threonine operon were derived from strain KY10935 by selection for auxotrophy for dipeptide L-alanyl-L-threonine or glycyl-L threonine, the transport systems of which were different from those of L threonine. All three independent mutants isolated needed an extraordinarily high concentration (10 mg/ml) of L-threonine, but grew in the presence of a low concentration (10 micrograms/ml) of either dipeptide, indicating that strain KY10935 had impaired L-threonine uptake. These results suggested that the strain had an unusual mechanism of L-threonine hyperproduction: the inability to take up L-threonine that had accumulated extracellularly decreased the steady-state level of intracellular L-threonine, freeing the remaining regulatory steps of feedback inhibition. PMID- 9404066 TI - Structural analysis of N-glycans of storage glycoproteins in soybean (Glycine max. L) seed. AB - The structures of N-linked sugar chains (N-glycans) of storage glycoproteins in soybean seeds have been identified. Eight pyridylaminated (PA-) N-linked sugar chains were derived and purified from hydrazinolysates of the storage glycoproteins by reverse-phase HPLC and size-fractionation HPLC. The structures of the PA-sugar chains purified were first identified by two-dimensional PA-sugar chain mapping and ion-spray mass analysis, considering the results of sugar composition analysis or sequential exoglycosidase digestion. The deduced structures were further analyzed by ion-spray tandem mass spectrometry and 500 MHz 1H-NMR spectrometry. The eight structures fell into two categories; the major class (96.6%) was a typical high mannose-type, the minor class was a xylose containing-type (Man3Xyl1GlcNac2, Man3Fuc1Xyl1GlcNac2; 3.4%). PMID- 9404068 TI - Activation of macrophages by sulfated glycopeptides in ovomucin, yolk membrane, and chalazae in chicken eggs. AB - Sulfated glycopeptides in ovomucin, chalazae and yolk membrane were found to activate cultured macrophage-like cells, J774.1, and TGC-induced macrophages from the peritoneal cavity of male mice. The macrophage-stimulating activity was estimated by the growth and morphology of the cells, H2O2 generation, and interleukin-1 (IL-1) production from the cells. The in vitro culture assay with macrophages showed that the protease digests of ovomucin, yolk membrane, and chalazae induced morphologic alteration and increased H2O2 generation and IL-1 production in lower concentration (100 micrograms/ml). The isolation of the components having macrophage-stimulating activity was attempted to elucidate the molecular mechanism. The O-linked carbohydrate chains, consisting of N acetylgalactosamine, galactose, N-acetylneuraminic acid and sulfate, in the sulfated glycopeptide were identified as a component having macrophage stimulating activity. PMID- 9404069 TI - Protective effect of green tea extract and tea polyphenols against the cytotoxicity of 1,4-naphthoquinone in isolated rat hepatocytes. AB - The cytoprotective effect of green tea extract and its phenolic compounds against 1,4-naphthoquinone-induced hepatotoxicity was evaluated in primary cultured rat hepatocytes. After exposure to 1,4-naphthoquinone, lactate dehydrogenase (LDH) leakage and cell viability were both improved by the presence of the tea extract and tea polyphenols. This cytoprotective effect was related to the structure of tea polyphenols, the galloyl group of (-)-epigallocatechin-3-gallate and (-) epicatechin-3-gallate being particularly effective. The production of liquid peroxidation by 1,4-naphthoquinone was not inhibited by the tea extract nor by tea polyphenol addition. After 2 h of incubation, the protein thiol concentration was reduced by 1,4-naphthoquinone, but this reduction was prevented by the tea extract and tea polyphenols. The reduction in protein thiol content of the cells closely paralleled the LDH leakage and loss of cell viability. These results suggest that the mechanism of protection by tea polyphenols against 1,4 naphthoquinone-induced toxicity to rat hepatocytes was due to the maintenance of protein thiol levels. PMID- 9404070 TI - Immunopotentiating activity of the water-soluble lignin rich fraction prepared from LEM--the extract of the solid culture medium of Lentinus edodes mycelia. AB - The water-soluble lignin in LEM (the extract of the solid culture medium of Lentinus edodes mycelia) has been known to have antiviral and immunopotentiating activities in vivo and in vitro. The water-soluble lignin rich fraction (JLS-18) was prepared from LEM using ultrafiltration and hydrophobic column chromatography. JLS-18 showed about 70 times higher antiviral activity than LEM in vitro. JLS-18 activated the cytotoxicity of NK cells and macrophages, and activated T cells in vitro. JLS-18 also induced interleukin 6 (IL-6) secretion from human leukocytes infected with Sendai virus in vitro. These data showed that JLS-18, the water-soluble rich fraction of LEM, had antiviral and immunopotentiating activities. PMID- 9404072 TI - Structural property and in vitro self-assembly of shark type I collagen. AB - The main structure of shark type I collagen is similar to that of land mammals, with a partial difference in amino acid sequence and post-translational modification. By static light scattering, the weight-average molecular weight of shark collagen (7.52 x 10(5)) suggests the presence of some aggregated molecules, oligomeric collagen. The self-assembly curve of shark collagen had a shorter lag phase and a longer growth phase than that of pig collagen. The optimum temperature and pH of shark collagen self-assembly is different from that of pig collagen. PMID- 9404071 TI - Enzymatic properties of double mutant enzymes at Asp51 and Trp49 and Asp51 and Tyr57 of RNase Rh from Rhizopus niveus. AB - Mutation of Asp51 of a base-nonspecific RNase, RNase Rh, to Ser, Thr, or Gln makes the enzyme more preferential for the dinucleoside phosphate (XpY) having G and C at the 5'-side (X). On the other hand the mutation of one of the B1 site components, Tyr57 to Trp, and Trp49 to Phe makes the enzyme more preferential for purine bases and pyrimidine bases, respectively. In this study, to obtain more specific RNases and RNases with different base specificity, we prepared double mutant enzymes that have Ser, Thr, and Asn at the 51st position and Trp at the 57th position or Phe at the 49th position, and their enzymatic specificities were studied with XpYs as substrates. The double-mutant enzymes D51SY57W and D51TY57W are more guanylic acid preferential than the mother single-mutant enzymes, D51S and D51T, respectively. They are extremely guanylic preferential RNases. D51NY57W is more a guanylic acid preferential enzyme than D51N, but cytidylic acid preference is of a similar order to that of D51N. The double mutant enzymes D51NW49F and D51TW49F showed an increased cytidylic acid preference as well as guanylic acid preference as compared to the mother single-mutant enzymes, D51T and D51N. The results of analysis of base specificity by the release of mononucleotides from RNA and the rates of hydrolysis of homopolynucleotides led to the same conclusion as in the case of the hydrolysis of XpY. PMID- 9404073 TI - Growth suppressing activity for endothelial cells induced from macrophages by carboxymethylated curdlan. AB - A carboxymethylated derivative of a linear (1-->3)-beta-D-glucan (CMCD) from Alcaligenes faecalis var. myxogenes acted directly on mouse peritoneal macrophages and mouse lymphoma P388D1 cells, and induced a growth suppressing activity for bovine artery endothelial cells (BAEs) from themselves at a concentration of 100 micrograms/ml. The suppressing activity was also detected in the mouse serum administered as an i.p. injection of CMCD at a dose of 100 mg/kg, suggesting that the growth suppressing activity was induced from macrophages potentiated by CMCD in vivo. PMID- 9404074 TI - Lipase-catalyzed synthesis of arbutin cinnamate in an organic solvent and application of transesterification to stabilize plant pigments. AB - Arbutin cinnamate was synthesized from arbutin (4-hydroxyphenyl beta-D glucopyranoside) and vinyl cinnamate by regioselective transesterification with a bacterial lipase in acetonitrile. The product was identified by NMR and FAB-MS analyses. These spectra showed that one ester bond was formed between the primary alcohol moiety of the D-glucose of arbutin and the carboxyl residue of cinnamic acid. Furthermore, plant pigments such as isoquercitrin (quercetin 3-O-beta-D glucopyranoside) and callistephin (pelargonidin 3-O-beta-D-glucopyranoside) were also converted to their corresponding cinnamate esters in the same manner. PMID- 9404075 TI - Heterodimeric aminopeptidase A from Bacillus licheniformis NS115. AB - An aminopeptidase A (EC 3.4.11.7) was purified to homogeneity from Bacillus licheniformis NS115 and its enzymatic properties were characterized. The enzyme had an apparent molecular mass of 64 kDa, consisting of heterodimeric 42 kDa and 22 kDa subunits, and is a new enzyme from N-terminal analysis of heavy and light subunits. The light subunit had no catalytic activity against the substrate and apparent K(m) values of heavy and whole enzyme were 0.26 and 0.087 mM of gamma glutamyl-p-nitroanilide, respectively. PMID- 9404076 TI - Stimulation of tumor necrosis factor and interleukin-1 productivity by the oral administration of cabbage juice to rats. AB - The effect of orally administering cabbage juice on tumor necrosis factor alpha (TNF) and interleukin-1 (IL-1) productivity was studied in resident peritoneal macrophages from normal and hepatoma-bearing rats. The productivity of TNF and IL 1 was stimulated by gastric intubation of cabbage juice in the normal state, but not in the hepatoma-bearing state where the production of these cytokines had already been stimulated. From these results, cabbage may contain some effective component(s) that can be absorbed from the gastrointestinal tract to stimulate the production of TNF and IL-1. PMID- 9404077 TI - An established hybridoma clone producing a monoclonal antibody against Vibrio anguillarum. AB - Vibrio anguillarum is a pathogenic microorganism of vibriosis, an infectious disease found in various fish species. A mouse hybridoma clone, named C5, that produced a monoclonal antibody to V. anguillarum was established. The specific reaction of C5 antibody with V. anguillarum was confirmed by the pre-adsorption effect of the V. anguillarum cells in ELISA and a cell immunoprecipitation experiment. Western blotting analysis indicated that the C5 antibody recognized a high molecular weight substance extracted from cells with detergents. PMID- 9404078 TI - Antioxidant activity of ferulic acid beta-glucuronide in the LDL oxidation system. AB - Antioxidant activity of ferulic acid beta-glucuronide, which is prepared from the plasma of feruloyl arabinose fed rats, in the CuSO4- induced LDL autoxidation system was studied. It has been found that absorbed ferulic acid occurred as the beta-glucuronide and that the antioxidant activity of ferulic acid beta glucuronide, which has not only the hydrophobic ferulic acid moiety but also a hydrophilic sugar moiety, is stronger than ferulic acid in the LDL oxidation system. PMID- 9404079 TI - Photocatalysis-dependent inactivation of Lactobacillus phage PL-1 by a ceramics preparation. AB - A ceramics preparation (Cleansand-205), which was coated with a mixture of the oxides of Si, Al, Ti, and Ag, was found to inactivate Lactobacillus phage PL-1 suspended in a buffer solution. The inactivation of phage was dependent on the amounts of Cleansand-205 added, and the reaction obeyed almost first-order reaction kinetics. The phage inactivation was considerably accelerated by the presence of light. PMID- 9404080 TI - Human placental fructose-6-phosphate,2-kinase/fructose-2,6-bisphosphatase: its isozymic form, expression and characterization. AB - The nucleotide sequence of 1981 bp cDNA containing the entire coding region of a human placental fructose-6-phosphate,2-kinase/fructose-2,6-bisphosphatase was determined. The sequence encodes 469 amino acids and, based on homology to the rat testis enzyme, appears to be the testis-type isozyme expressed in placenta. The enzyme was expressed in Escherichia coli BL21 (DE3) by using a T7 RNA polymerase-based expression system and purified to homogeneity. The expressed enzyme was bifunctional with specific activities of 75 and 80 mU/mg of kinase and phosphatase, respectively. Kinetic parameters of the expressed enzyme are similar to those of the rat testis enzyme. PMID- 9404081 TI - Nitric oxide formation by macrophages stimulated with water extracts from meats and offals. AB - Water extracts from meats and offals were incubated with a macrophage cell line (RAW 264.7), and nitrite in the medium was measured as an index of the macrophage stimulating activity. Ten of 38 water extracts had macrophage stimulants and chicken meat, chicken gizzard, cattle reticulorumen, swine stomach, and swine cerebrum had high activities. PMID- 9404082 TI - Synthesis of asymmetrically labeled sucrose by a recombinant sucrose synthase. AB - About 80% of radioactivity was recovered in asymmetrically labeled sucrose from UDP-[14C]glucose or [14C]fructose with recombinant mung bean sucrose synthase expressed in Escherichia coli harboring pEB-01. This high recovery is due to the fact that the enzyme conserving the activity of sucrose synthase has a similar affinity for UDP-glucose and fructose to an intact enzyme from the mung bean, but a lower affinity for sucrose. PMID- 9404083 TI - Biodegradabilities of ethylenediamine-N,N'-disuccinic acid (EDDS) and other chelating agents. AB - Biodegradabilities of chelating agents were tested with activated sludge. Ethylenediaminetetraacetic acid (EDTA) remained intact in the effluent even after acclimation for 100 days, but propanediamine-N,N'-disuccinic acid (PDDS) and nitrilotriacetic acid (NTA) were biodegraded after acclimation for 5 and 23 days, respectively. Optical isomers of ethylenediamine-N,N'-disuccinic acid (EDDS) had different biodegradabilities: SS- and RS-isomers were susceptible to biodegradation, but the RR-isomer was resistant. SS-isomer was degraded even by activated sludge without acclimation. PMID- 9404084 TI - Panton-valentine leukocidin genes in a phage-like particle isolated from mitomycin C-treated Staphylococcus aureus V8 (ATCC 49775). AB - The staphylococcal Panton-Valentine leukocidin (PVL) genes [lukS-PV-lukF-PV] existed in a hexagonal phage-like particle (phi PVL) isolated from mitomycin C induced Staphylococcus aureus V8 (ATCC 49775). The genome packed in phi PVL was a linear double-stranded 40-kb DNA with single-stranded cohesive ends (cos). The [lukS-PV-lukS-PV], attP, and int (integrase gene) of phi PVL were all located very close to one another within a 4.0 kb-segment on the genome in the order given, and the segment is located at the center from the left and the right cos sites. In addition, the [lukS-PV-lukF-PV]-attP-int region contains 5 direct repeat sequences that show high similarity with the recombinase-binding sites of bacteriophages of S. aureus. PMID- 9404085 TI - [Unknown episodes on IgE]. PMID- 9404086 TI - [Clinical practice of childhood asthma]. PMID- 9404087 TI - [Biology of mast cells ad basophils]. PMID- 9404088 TI - [Assessment of allergenic activity of heated and ovomucoid-depleted egg white]. AB - We investigated allergenic activity of heated and ovomucoid (OM)-depleted egg white by RAST inhibition tests, skin tests and and oral challenge tests. Freeze dried egg white, OM and ovalbumin (OA) were coupled with CNBr-activated paper discs. Freeze-dried, heated, and heated and OM-depleted egg white antigens were used as inhibitors in RAST inhibition tests. Freeze-dried egg white significantly inhibited the IgE-binding to freeze-dried egg-white-, OM- and OA-discs. Heated egg white showed a significant inhibition against only OM-disc, but heated and OM depleted egg white didn't effectively suppress IgE-binding to all discs. Fifty six patients were subjected for skin tests and 27 patients for oral challenge tests with freeze-dried egg white, heated egg white and heated and OM-depleted egg white. Eighteen of 25 subjects with positive prick tests results to freeze dried egg white showed negative results to heated egg white. Four of 7 cases with positive prick test results to heated egg white gave negative results to heated and OM-depleted egg white. 20-minutes patch tests showed almost similar results to prick tests. Four of 8 subjects showing positive oral challenge tests with freeze-dried egg white were negative to heated egg white. All of 4 with positive oral challenge tests by heated egg white were negative results to heated and OM depleted egg white. These evidence indicated that heated and OM-depleted egg white was more hypoallergenic than heated egg white. PMID- 9404089 TI - [Effect of vacuum cleaning of room floors and bed clothes of patients on house dust mites counts and clinical scores of atopic dermatitis. A double blind control trial]. AB - By a randomized double blind control trial we studied the effect of vacuum cleaning of room floors, mattresses and quilts for twelve months on clinical symptoms and laboratory data of atopic dermatitis patients. All patients used the identical new vacuum cleaners. Thirty patients (3-12 years of age) with relatively stable skin conditions were randomly allocated to either of the following two groups. In the monitor group we visited patient's home every three weeks and mite specialists cleaned drastically the room floors, mattresses and quilts and the patient continued to clean in the same way in-between. In the control group we visited similarly but the cleaning was made insufficiently which was also followed by the patient. But, at 2 occasions (the first and the last visits), cleaning was made drastically also in the control group. Thus the mite numbers were counted precisely at the start and the end of the experiment both in the monitor and control groups. Each patient was seen every six weeks by the same doctor and estimated of his symptoms using our unique scoring system (Fig. 1). At the start and the end of the study, total IgE and specific IgE antibodies to house dust mites in the serum were evaluated. The monitor group showed a tendency to count smaller number of mites than the control group after a year, when there was a significant difference only in quilts. However, a statistically significant decrease in the mite counts was observed only in room floors and not in mattresses and quilts both in the monitor and control groups (Fig. 2). Clinical scores after a year significantly improved only in the monitor group and not in the control group (Fig. 3). Serum IgE value and specific antibody titer to house dust mites were not changed significantly after the trial in both groups. As a conclusion, vacuum cleaning of the patient's room improved the clinical symptoms of atopic dermatitis but this could not be related to the reduction of house dust mite number. PMID- 9404090 TI - [A epidemiologic study on the prevalence of the allergic diseases in school children in Kyoto City]. AB - The prevalence of allergic diseases in school children in Kyoto City (17,906 children) was examined with the questionnaire which was prepared by the Study Group of Epidemiology of Allergic Diseases funded by the Ministry of Public Welfare in 1993. RESULTS: 1) The number of valid reply was 16,176 (reply rate 90.3%). The reply rate of each question was from 89.5% to 98.9% (average 95.7%). 2) The prevalence of bronchial asthma was 4.9% in the elementary school children, 3.5% in the junior high school children and 4.5% in total, and the prevalence in boys was higher than girls (ratio 1.6). The prevalence of wheezing was 6.5%, 2.7% and 5.5%, respectively, and the sex ratio was 1.2. 3) The prevalence of atopic dermatitis was 5.3% in elementary school children, 4.0% in junior high school children and 5.0% in total, and the prevalence in boys was lower than girls (ratio 0.87). 4) The prevalence of allergic rhinitis was 19.8% in elementary school children, 21.2% in junior high school children and 20.3% in total, and the prevalence in boys was higher than girls (ratio 1.3). 5) The prevalence of allergic conjunctivitis was 11.9% in elementary school children, 15.4% in junior high school children and 13.0% in total and the prevalence in boys was almost same as girls (ratio 0.98). 6) The prevalence of any allergic diseases was 32.2% in elementary school children, 31.7% in junior high school children and 32.1% in total and the prevalence in boys was higher than girls (ratio 1.2). PMID- 9404091 TI - [The comparative study of LUMIWARD immunoassay system and CAP RAST system for determination of food allergy on infantile atopic dermatitis]. AB - The reliability of assay methods was studied between LUMIWARD and CAP RAST due to the removal of some kinds of foods and/or challenge test. In the 45 infants with atopic dermatitis, egg, milk, flour, rice, and soybean were removed from foods and/or challenge test of those foods were carried out by every three months as possible. At the same time, the allergen-specific IgE antibodies were measured with LUMIWARD and CAP RAST. The negative concordance rate of LUMIWARD and CAP RAST. The negative concordance rate of LUMIWARD was elevated more than that of CAP RAST and the positive concordance rate of CAP RAST was elevated more than that of LUMIWARD. These results suggests that CAP RAST is useful for the determination of removal of foods, while LUMIWARD is useful for the determination of foods to challenge. The risk of overlooking the allergen exists, because the positive concordance concordance rates of LUMIWARD were low in the milk and flour. With developing age, the negative concordance rates decreased in CAP RAST, therefore unnecessary removal of foods may continue for a long time. The determination of low concentration with LUMIWARD was unreliable. PMID- 9404092 TI - [Preventative effect of a whey hydrolyzed formula (Nestle, NAN H.A.) on the development of allergic symptoms in infants]. AB - We examined the prevent effect of a whey hydrolyzed formula (NAN H.A.) on the development of allergic disease from new born infants. A hundred thirty three pregnant women were divided into the NAN H.A. group and the control group. Mothers, who agreed to use the whey hydrolyzed formula to their infant, were classified into the NAN H.A. group. We examined laboratory date such as IgE levels of these infants as well as clinical symptoms for evaluating of the preventive effect of NAN H.A. The serum IgE levels from infants of the NAN H.A. group were lower than those of the control group. The incidance of the infants who showed allergic clinical symptoms was significant lower in the NAN H.A. group. The odds ratio to skin symptom at 3 years of age with multiple logistic regression was 5.32 between the control group and the NAN H.A. group (p < 0.05). It was 6.68 between mother's history of allergy was positive and negative (p < 0.02). These results suggest that NAN H.A. can prevent the development of allergic symptoms in infants. PMID- 9404093 TI - [Comparative studies of airway deposition of 99mTc-DTPA aerosol particles among 4 different nebulizers by pulmonary scintillation method]. AB - The inhalation therapy is one of the important treatment of bronchial asthma. The effectiveness of the therapy, however, is different depending on the size of particles produced by each inhalation equipment. Therefore, we have tried to clarify the relationship between the size of inhaled particles and the deposition rate in the lung by using different types of equipment. Four different equipments, Nishou, Pari-Boy, Pari-Master and Atom Soniclizer 905 were used. 99mTc-DTPA was dissolved in physiological saline and was inhaled for evaluation of "deposition" rate in the lung. The deposition rates by Nishou, Pari-Boy, Pari Master and Atom Soniclizer 905 were 10.4 +/- 5.8%, 27.5 +/- 14.5%, 28.5 +/- 6.5%, and 31.1 +/- 6.8%, respectively. Most of 99mTc-DTPA inhaled was observed in oral cavity. The deposition in the lung by Nishou was significantly reduced compared to other three nebulizers. Those three were evaluated as appropriate equipments for the treatment. The average size of particles in an airsol by those three equipments was less than 5 microns. The study suggests that the size of "particles" in an airsol is very important factor for the inhalation therapy. PMID- 9404094 TI - [Clinical effect of alprazolam and trazodone on bronchial asthmatics with anxiety disorder]. AB - Onset as well as many symptoms of asthma are said to be related to autonomic dysfunction which often appear as a result of actual daily stress of the patients. Minor tranquilizers and anti-depressants are often successful in treating such symptoms, though precise effect of these drugs on asthma is not yet available, while many still hesitate to use these drug for fear of the anticholinergic effect which may hinder the efficient expectoration of sputum which could be critical during severe asthma attacks. In this study, these drugs were evaluated by the asthma score, peak flow, acetylcholine provocation test and the suppressive effect on lymphocytes. They could be beneficial in increasing the quality of asthma therapy if employed with necessary care. PMID- 9404095 TI - [18 cases of asthma induced by hamster or guinea-pig bred as pets]. PMID- 9404096 TI - [Study on the evaluation of total and regional liver function using 99mTc-GSA dynamic SPECT]. AB - To evaluate regional and total liver function, 99mTc-GSA dynamic SPECT was studied in 58 patients with various liver disease and 5 normal volunteers. Using dual-head gamma camera, 60 projection data (90 seconds/rotation) were acquired to obtain SPECT imaging. Forty continuous SPECT were obtained for 60 minutes. Time activity curve (TAC) of each voxel was created. The TAC was assumed to be equally the function of C(t) = Cmax (l-e-kt). The liver uptake rate (K value) was calculated using the least squares method. The product of K value and functional volume of each voxel was defined as liver functional index and the sum of the indices was defined as total liver functional index. The total liver functional index had good correlation with various liver function test, HH15, LHL15, and the index showed significant difference between each group of Child-Pugh's liver dysfunction stage. Because of unnecessariness of setting up the ROI, this method is simple and the result is no difference among operators. Three-dimensional liver functional index map exactly shows regional liver function. So that, this method seems to be useful for predicting the residual liver function after hepatectomy. PMID- 9404097 TI - [Quantification of brain perfusion SPECT with N-isopropyl-p-iodoamphetamine using noninvasive microsphere method: estimation of arterial input by dynamic imaging]. AB - We have developed a noninvasive method to quantify brain perfusion SPECT with 123I-N-isopropyl-p-iodoamphetamine (IMP) using serial dynamic planar imaging of the initial transit phase. The method is based on the microsphere model, but does not require arterial sampling. Serial dynamic planar imaging was performed for 6 min after the bolus injection of IMP (167 MBq in 1.5 ml), followed by additional planar imaging at 20 min and SPECT scan thereafter. The total arterial input to the brain during the initial 5 min after injection was estimated by the injected dose, with the correction of the lung retention, divided by cardiac output (CO). CO was estimated from the initial transit of IMP in the right heart. Cardiac output index (COI), obtained from the integral of the first transit of IMP in the right heart divided by the injected dose, was calibrated by CO measured by Doppler ultrasonography. Regional cerebral blood flow (rCBF) obtained by this method in normal subjects was acceptable. However, the results may be influenced by the injection technique, and careful attention should be considered for clinical application of this method. PMID- 9404098 TI - [Reduction in mean cerebral blood flow measurements using 99mTc-ECD-SPECT during normal aging]. AB - Mean cerebral blood flow (mCBF) was measured by SPECT using the 99mTc-ECD-Patlak Plot method in a selected group of 61 normal non-hospitalized subjects aged 51 to 91 years. The mCBF values showed 48.4 +/- 4.7 ml/100 g/min in aged 50-59 years group, 49.9 +/- 5.9 ml/100 g/min in aged 60-69 years group, 46.4 +/- 6.5 ml/100 g/min in aged 70-79 group, 38.0 +/- 3.7 ml/100 g/min in aged 80-89 years group, 38.9 ml/100 g/min in aged 90-99 years group. There was a statistically significant reduction of mCBF with advancing age (R = -0.41; p = 0.001). Women have significantly higher mCBF values than men up to aged 70 years group. In this study, there was no significant laterality in the mCBF between right and left hemispheres in all decade groups. The history of hypertension, alcohol consumption, and cigarette smoking failed to show significant difference in the mCBF values. The present study shows that normal aging is associated with mCBF reduction. PMID- 9404099 TI - [Assessment of 123I-beta-methyl iodophenyl pentadecanoic acid (BMIPP) myocardial scintigraphy in patients of chronic right ventricular overload--fatty acid metabolism in right ventricular myocardium]. AB - An investigation on the right ventricular pressure level and the abnormalities in the fatty acid metabolism of myocardium was made using 123I-beta-methyl iodophenyl pentadecanoic acid (BMIPP) myocardial SPECT in patients with chronic right ventricular overloading. Twenty patients who presented with right ventricular systolic pressure (RVSP) of 35 mmHg or more were used as the subjects. Dual myocardial SPECT with 201TlCl (Tl) and BMIPP was carried out for the subjects and RVc/LVc, a ratio of radioactivity count incorporated in the right ventricular free wall to the left one was determined for Tl and BMIPP. And the correlations between RVc/LVc and RVSP, and RVc/LVc and RVSP/LVSP were examined. The subjects were classified into 3 groups based on the RVSP levels and the count ratio, BMIPP/Tl was compared among the three groups. With respect of Tl uptake, there were significant, positive correlations between RVc/LVc and RVSP (correlation coefficient r = 0.51, p < 0.05) and between RVc/LVc and RVSP/LVSP (correlation coefficient r = 0.59, p < 0.01). On the other hand, no significant correlation was found between them with respect of the uptake of BMIPP. The BMIPP/Tl ratio in the group with higher than 80 mmHg of RVSP was 0.82 +/- 0.06, which was significantly lower than the ratio's for two groups of less than 80 mmHg; 0.91 +/- 0.07 and 0.98 +/- 0.04 in the group with 35-49 and 50-79 mmHg of RVSP, respectively. These results show that when compared with BMIPP, Tl is superior for the estimation of right ventricular pressure. For the patients with right ventricular overloading, it was suggested that when RVSP reaches 80 mmHg or more, there appear some disorders in the fatty acid metabolism in the right ventricular myocardium. PMID- 9404100 TI - [Successful TSH suppression therapy with triiodothyronine in a patient with pulmonary metastases from differentiated thyroid carcinoma in the absence of 131I uptake]. AB - A 28-year-old woman was referred to us to undergo 131I therapy who had multiple pulmonary metastases from papillary thyroid carcinoma after total thyroidectomy. There was no increased accumulation of a tracer in the pulmonary metastatic foci on whole-body scanning using a 111 MBq diagnostic dose of 131I. However, the pulmonary metastases were gradually decreased in size, and then clearly reduced 8 months after the start of TSH suppression therapy, which was maintained by T3 instead of T4 to bring down the serum TSH level below 0.1 microU/ml. Reduction rates of the foci were 33-76% on chest X-ray. The reduction was confirmed and no new lesions were found on the serial CT scans. Serum thyroglobulin level was lowered 80 to 25 ng/ml by this suppression therapy and progression of disease was not observed under a 54 months' T3 treatment. Thus, TSH suppression therapy might improve survival of patients with differentiated thyroid cancer. PMID- 9404101 TI - [Pitfall in 201Tl-99mTc subtraction scintigraphy: a case of 99mTc-pertechnetate uptake in a parathyroid hyperplasia]. AB - 201Tl-99mTc subtraction scintigraphy has been recognized as a useful procedure in the preoperative localization of hyperfunctioning parathyroid glands. We experienced a case which showed 99mTc-pertechnetate uptake in a parathyroid hyperplasia. This case warned us to focus a lot of attention on the detection for preoperative localization. There has been no such case reported in the previous Japanese literatures. Hypervascularity and thick fibrous capsule presumed explanation for a rare case of marked pertechnetate uptake into a parathyroid hyperplasia. PMID- 9404102 TI - [Comparative study on the tumor accumulation of 99mTc-tetrofosmin and 99mTc-MIBI in rabbits bearing VX-2 cancer]. AB - Each of myocardial blood flow imaging agents has a potential usefulness as an agent for tumor scintigraphy. The tumor accumulation and washout of 99mTc tetrofosmin and 99mTc-MIBI were comparatively studied using rabbits bearing VX-2 cancer. From seventeen to twenty days after the implantation of VX-2 cancer into the femoral region of seven rabbits, tumor to soft tissue accumulation ratio (T/S ratio) of each agent was calculated in early images (5 min after injection) and in late images (50 min after injection). Compared with 99mTc-tetrofosmin, the T/S ratio of 99mTc-MIBI was higher and, moreover, the washout was delayed. These results suggest that there is a difference in tumor accumulation property between these two agents. PMID- 9404103 TI - [Emergent operation after percutaneous transluminal coronary rotational atherectomy (ROTABLATOR)]. AB - Emergent operations were performed in seven patients after percutaneous transluminal coronary rotational atherectomy (PTCRA). The causes of the emergent operations were coronary rupture in three patients, acute coronary occlusion in two patients, perforation of the ascending aorta in one patient and impossible weaning from IABP in one patient. IABP was used preoperatively in all patients. Coronary artery bypass grafting was performed in all patients. Ruptured sites of the coronary arteries were closed and perforated site of the ascending aorta was repaired. Two patients died due to cardiac failure but five patients were recovered. Coronary artery rupture was main complication after PTCRA. It is important to recognize the difference between the complication after PTCRA and that after conventional PTCA. PMID- 9404104 TI - [Wide resection and reconstruction of chest wall: usefulness of A-O metal plate]. AB - A 53-year-old male was admitted to our department with a chest tumor due to infiltration of lung cancer. Right upper lobectomy and a wide en bloc excision of the right upper chest wall were performed, including the 2nd, 3rd, 4th, and 5th ribs with upper lobectomy of right lung. The defect was replaced with three A-O metal plates, which joined both stumps of the ribs, from the third to the fifth rib. These metal plates were very useful because it was easy to bent and twist them to fit the defect in operation. And curved metal plates preserved a cone from of chest cage. Postoperative course was favourable without mechanical ventilation and wound infection. PMID- 9404105 TI - [Surgical relief of airway obstruction from a double aortic arch associated with corrected transposition of the great arteries, pulmonary atresia and bilateral patent ductus arteriosus in a neonate]. AB - A rare 20-day-old male with double aortic arch, corrected transposition of the great arteries (cTGA), pulmonary atresia and bilateral patent ductus arteriosus (PDA) was transported to our institute because of severe respiratory dysfunction and cyanosis. The patient had been already intubated and ventilated on respirator. A echocardiography and cine-angiography demonstrated that the both sides aortic arch had almost identical sizes, originating common carotid arteries and subclavian arteries and PDAs respectively, and the descending aorta located on the left side of the mid-line. At the first surgery, the distal of the right aortic arch was divided just proximal to the descending aorta after complete tissue dissection around the arch. The divided right sided aortic arch was mobilized from posterior to anterior aspect of the bronchus. Then the right subclavian artery was divided and an original Blalock-Taussig shunt was employed. The right sided PDA was ligated. After the first surgery, respiratory dysfunction lasted for weeks mainly because of the PGE1 dependent left sided PDA. At the second surgery, left sided modified Blalock-Taussig shunt was constructed and the left sided PDA was divided. These procedures resulted in stable respiratory status and oxygen saturation. The patient was extubated three days later and now in satisfactory clinical condition. PMID- 9404106 TI - [Clinical effect of high-flow pulsatile cardiopulmonary bypass in coronary artery bypass grafting]. AB - The effect of high-flow pulsatile cardiopulmonary bypass was evaluated in 36 patients undergoing coronary artery bypass grafting in our unit. The patients were divided into two groups, based on cardiopulmonary bypass (CPB) flow; high (3.0 +/- 0.2 l/min/m2), or moderate (2.4 +/- 0.2 l/min/m2). Multidose cold crystalloid cardioplegia was administered for myocardial protection. Pulsatile flow during CPB was used and systemic perfusion pressure was maintained between 50 and 80 mmHg. Preoperatively, there were no differences between groups in left ventricular ejection fraction or extent of coronary artery disease. The times required for CPB and weaning from CPB were significantly shorter in high-flow group than moderate-flow group. The urinary output during CPB was significantly higher in high-flow group than moderate-flow group. Postoperatively, there were no significant differences in the incidence of myocardial infarction, stroke, or 30-day mortality between groups. In conclusion, high-flow pulsatile CPB shortens the length of CPB and does not differ significantly from moderate-flow with respect to mortality and morbidity. PMID- 9404107 TI - [A simple device of chest wall reconstruction]. AB - We devised a simple method of chest wall reconstruction in two cases of malignant tumor of the chest. We strained a suture (adsorbable or monofilament) to the intact ribs above and below the defect and fixed the sheets of Marlex mesh in double layers and closed the skin without any myocutaneous flap. The postoperative course was uneventful. This device is simple and effective method to maintain the stability of the chest wall defect. PMID- 9404108 TI - [MIDCAB (minimally invasive direct coronary artery bypass) using mini-CABG instruments: a case report]. AB - A 61-year-old man with unstable angina due to complete occlusion of the proximal LAD underwent MIDCAB (left internal thoracic artery (ITA)-left anterior descending artery (LAD) anastomosis), because of active hepatitis and unsuccessful PTCA. The left fourth intercostal mini-thoracotomy (9 cm long) was performed. The left ITA was harvested through the thoracotomy. The left ITA to LAD anastomosis under the beating heart was successfully performed using Mini CABG instruments (USSC). Postoperative recovery was uneventful and postoperative coronary angiography on the 5th postoperative day revealed a widely patent graft. The patient was discharged on the 8th postoperative day. This device was very useful to perform the MIDCAB procedure. PMID- 9404110 TI - [Case performed: simultaneous surgery for left coronary ostial stenosis and gastric cancer]. AB - This report describes a case in which a 68-year-old male underwent two operations simultaneously for left coronary ostial stenosis and gastric cancer. Successfully performed procedures were a single coronary artery grafting with the saphenous vein to the left anterior descending artery, and a subtotal gastrectomy using the Billroth II method. The postoperative course was uneventful and the patient was discharged from the hospital in good condition after 42 days. At present, one year postoperative, the patient has been visiting the outpatient clinic in healthy condition. PMID- 9404109 TI - [Reoperation for mitral regurgitation 13 years after aortic valve replacement and manouguian's anulus enlargement: report of a case]. AB - The patient was 22-year-old female. She had undergone aortic valve replacement and Manouguian's anulus enlargement with low porosity woven Dacron patch for congenital aortic stenosis 13 years ago, and developed mitral regurgitation 9 years after that operation. Two regurgitant flow were observed. One was originated from the orifice due to mitral prolapse. The other was from a tear in the anterior leaflet. It was around the tip of the prosthetic patch, approximately 7 mm in size, and was repaired easily. But the mitral valve itself was found to be malformed and prolapsed, requiring mitral valve replacement. Her postoperative course was uneventful. PMID- 9404111 TI - [A case report of coronary artery bypass grafting in a patient with ankylosing spondylitis]. AB - We reported a case of ankylosing spondylitis who successfully underwent coronary artery bypass grafting (CABG) for unstable angina pectoris. A 67-year-old man was admitted with symptom of anginal pain. Selective coronary angiography revealed coronary artery stenoses; 90% in seg 6, 90% in seg 11, proximal 75%, distal 90% in seg 3, 99% in 4 PD and 99% with delay in 4 AV. The left internal thoracic artery was anastomosed to seg 7 and saphenous vein (SVG) to PL-2, PL-1 sequentially, and another SVG to 4 PD. His postoperative course was uneventful. Cardiac lesions accompanied by ankylosing spondylitis are rare in Japan. Perioperative problems of these lesions therefore, are discussed. PMID- 9404112 TI - [Surgical treatment for endocardial cushion defect in an elderly patient with pulmonary hypertension]. AB - We performed surgery for a partial endocardial cushion defect (ECD) in a 67-year old female patient with pulmonary hypertension. The patient had grade 2 tricuspid regurgitation and pulmonary hypertension (Pp/Ps = 0.58, Rp = 8.4 Um2). The operative procedure involved patch closure of the ostium primum and tricuspid annuloplasty with a Carpentier ring. The postoperative course was uneventful and the pulmonary hypertension improved (Pp/Ps = 0.33). Surgery for partial ECD should be recommended even for elderly patients. PMID- 9404113 TI - [A Rastelli operation with a reconstruction of the central pulmonary artery for a pulmonary atresia with an absent left pulmonary artery]. AB - A Rastelli procedure was successfully performed on a 6-year-old girl with an absence of the intrapericardial pulmonary artery and the left pulmonary artery (PA), following a right B-T shunt. The central PA was reconstructed with an 18 mm diameter xenopericardial roll behind the ascending aorta and the superior vena cava. After intracardial repair, the Rastelli operation was performed using an 18 mm diameter composite graft which consisted of a valved xenopericardial roll and a knitted Dacron graft. Since the size of the right PA was large enough to undergo this surgical procedure (the preoperative right PA index was 300 mm2/m2), the postoperative peak systolic pressure ratio of the right ventricle to the left ventricle declined from 1.0 to 0.64. Although the patient showed slight signs of right ventricular failure on the operative day, the postoperative course was uneventful. PMID- 9404114 TI - [Coronary artery bypass grafting in a patient with a permanent tracheal stoma]. AB - The performance of open heart surgery in a patient with a tracheostoma can present difficult problems, including mediastinitis and inadequate operative exposure. A 79-year-old man was admitted because of angina pectoris, and had undergone tracheostomy for carcinoma of the larynx 14 years previously. He underwent coronary artery bypass grafting with saphenous vein graft to # 8, 12. A skin incision was made from the angle of Louis to the xiphoid process and carried down through the subcutaneous tissue. The sternum was divided from the xiphoid process to the manubrium and then dislocated from the intact manubrium. Operative time was 165 minutes and arrest time was 64 minutes. Postoperative course was satisfactory and discharged within 24 days after operation. We think that in a case of open heart surgery with a tracheostoma no dissection near a tracheostoma is necessary in order to decrease the risk of postoperative wound infection and mediastinitis. PMID- 9404115 TI - [A case of esophagopleural fistula successfully cured by conservative therapy after middle and lower lobectomy of right lung]. AB - We experience a case of esophagopleural fistula successfully cured by conservative therapy after the lung cancer operation. A 46-year-old man was received middle and lower lobectomy for adenoid cystic carcinoma of the right lung. Complication of empyema associated with an esophagopleural fistula occurred on postoperative 4th day. Conservative therapy of nothing by mouth, intravenous hyperalimentation and antibiotics was started. Three thoracic drains were inserted and the thoracic irrigation of total 3,000 ml warm saline per day twice on one day was continued. The esophagopleural fistula was closed on 6th week and the patient was discharged on 11th week after the therapy start. This complication is much rare, but recent advance in the diagnostic methods seemed to increase the indication of conservative therapy in future. PMID- 9404116 TI - [A case of medullary carcinoma of the thyroid gland: metachronous bilateral metastasis of the neck and mediastinal lymph nodes were treated successfully by staged resection]. AB - This patient, a 52-year-old male, underwent subtotal thyroidectomy on the diagnosis of medullary carcinoma of the thyroid gland in 1980 and postoperative course was uneventful. Since November 1990 he had a persistent diarrhea for 6 months and was admitted to the hospital for the further examination on June 1991. The serum CEA and calcitonine level was very high and chest CT scan findings showed the swelling of right neck and mediastinal lymph nodes. Dissection of the lymph nodes was performed by anterior approach which was gained through a proximal median sternotomy extended into the anterior fourth intercostal space as well as to the base of the neck on the right side. On the pathological examination it was metastasis of medullary carcinoma of the thyroid gland. And 50 months later after second operation he had a persistent diarrhea once again. Left neck and mediastinal lymph node metastasis was detected by chest CT with high serum CEA and calcitonine level. Similarly resection was performed by the same anterior approach on the left side. Irrespective of the extended resection he was free of severe complication; he is still alive 10 months after the third operation without any evidence of recurrence and his current performance status is very good. PMID- 9404117 TI - [A long-term surviving case with invasive thymoma undergone 4 times operations for tumor recurrences]. AB - We reported a long-term survivor with invasive thymoma associated with myasthenia gravis. The patient underwent operations 4 times for these 13 years. At the third and fourth operation, the tumor was resected after preoperative neo-adjuvant chemotherapy with cisplatin and adriamycin, and postoperative radiotherapy was added. Her postoperative course was uneventful, and she has been alive. Our strategy for invasive thymoma is that tumors which include both primary and recurrent lesions are removed as completely as possible, and that postoperative radiotherapy is added for stage II, III and IV in Masaoka's classification, and preoperative neo-adjuvant chemotherapy for stage III and IV. We believe that these multidisciplinary treatments and long term follow-up lead good results without harm to quality of life. PMID- 9404118 TI - [Sleeve lobectomy for tuberculous bronchial stenosis: a case report]. AB - We describe a patient with tuberculous bronchial stenosis who was subjected to bronchoplasty. The patient was a 33-year-old man who had stenosis of the left main bronchus. Because the lesion was associated with bronchomalacia, previous balloon dilatation therapy had failed. At thoracotomy, the left upper lobe was found not to be saved for the tuberculous lesion. Although there were many inflamed nodules in the left lower lobe due to repeated episodes of pneumonia, we decided to save it using bronchoplasty expecting its respiratory functional recovery. He ran uneventful course postoperatively and his lung function improved. We conclude that bronchoplasty may prove effective for patients with tuberculous bronchial stenosis associated with bronchomalacia; and thus, to avoid pneumonectomy, bronchoplasty should be attempted even if the reconstructed lung is mildly inflamed. PMID- 9404119 TI - [Redo operation for recurrent pulmonary artery aneurysm associated with pulmonary stenosis and regurgitation]. AB - A 63-year-old woman who underwent surgical correction of a recurrent pulmonary artery aneurysm associated with pulmonary stenosis and regurgitation is reported. On April 1986, she underwent commissurotomy of pulmonary valve, reconstruction of right ventricle out flow tract using a Polystan monocusp patch and pulmonary aneurysmorrhaphy for pulmonary artery aneurysm. Pathological examination of the resected pulmonary arterial wall revealed mucoid degeneration of media and fragmentation of elastic fiber. Nine years after the operation, recurrence of pulmonary artery aneurysm, pulmonary stenosis and regurgitation were recognized. On September 1995, she underwent redo operation with graft replacement of pulmonary artery and pulmonary valve replacement using woven Dacron prosthesis containing a Carpentier-Edwards bioprosthetic valve. We should choose as an initial procedure with graft replacement for pulmonary artery aneurysm with fragility of the pulmonary arterial wall. She is now doing very well at one year and 5 months after the redo operation. PMID- 9404120 TI - [A case of invasive thymoma that recurred with pericardial effusion 15 years postoperatively]. AB - We treated a 74-years-old male who had recurrence with pericardial effusion and a tumor between the pulmonary artery and left atrium 15 years after operation for invasive thymoma complicated by myasthenia gravis. Despite pericardial drainage, CDDP infusion, and radiation therapy for the tumor, the patient died due to pneumonia after 1 and half years. Long-term observation is considered to be needed after surgical resection of thymoma for recurrence or multiple oncogenesis in ectopic thymus. PMID- 9404121 TI - [An elderly case of pneumothorax treated with omentopexy]. AB - A 74-year-old male was admitted to our hospital because of left pneumothorax with persistent air leakage. He had undergone negative pressure drainage, chemical pleurodesis and transbronchial embolization in another hospital. Chest X-ray and CT scan showed collapse of the left lung and a defect of the pleura in the left lung S9. Patch closure was performed for the round pleurobronchial fistula (35 x 35 mm in size) using polyglycol acid felt and fibrin glue. But as severe air leakage was observed again about 24 hours after surgery, omentopexy was performed. The postoperative course was uneventful, and he was discharged 17 days after the second surgery. PMID- 9404123 TI - [Treatment of ARDS--present status and an outlook for the future]. PMID- 9404124 TI - [Aspiration pneumonitis; progress in understanding its acute pathophysiology and its therapy]. AB - Aspiration of gastric contents has been a major cause of acute respiratory failure in adult patients, and its mortality has been very high. Current method of treatment is limited, but the pathogenesis of acid aspiration lung injury has been well studied. The lung injury can be divided into direct and secondary injury. Neutrophils are thought to be a primary mediator of the secondary lung and systemic organ injury. Various substances such as cytokines, complements, and arachidonic acid metabolites, which activate neutrophils, are produced in acid aspiration pneumonitis. In this article, the progress in research of the acute pathophysiology of acid aspiration pneumonitis is reviewed and the possibility of its application to therapy is discussed. PMID- 9404125 TI - [The effect of hyperbaric oxygenation on nitrite and nitrate production in rats treated with endotoxin]. AB - The effect of hyperbaric oxygenation (HBO; 3 ATA, 90 min) on production of stable endproducts of nitric oxide (NO); nitrite (NO2-) and nitrate (NO3-), in plasma was studied in anesthetized rats treated with lipopolysaccharide (LPS). Intravenous injection of LPS (1 mg.kg-1) increased NO2-/NO3- production in plasma significantly 5-6 hrs after treatment, compared with non-treated rats. Production of NO2-/NO3- was not influenced by HBO performed 3 hrs after LPS injection. HBO performed 1 hr after LPS treatment, however, depressed NO2-/NO3- production significantly, compared with HBO performed 3 hrs after LPS, although NO2- production remained increased. These data suggest that HBO may influence the NO2 /NO3- production in NO producing processes in rats treated with LPS. PMID- 9404126 TI - [The effects of isoflurane on the formalin-induced activity in the spinal dorsal horn of transected cat]. AB - Although the formalin test is widely used in rodents, the electrical response to this stimulus and the effect of isoflurane on this response, has not been well understood in cats. We have therefore examined the effect of isoflurane on spinal wide-dynamic-range neuronal activity evoked by a subcutaneous (s.c.) formalin injection in cats. In decerebrate, spinal cord-transected cats (L 1-2), a s.c. injection of formaldehyde solution (0.05 ml; 5%) was performed in the receptive fields of spinal wide-dynamic-range neurons at the hind paw. Isoflurane (1.5%) inhalation was given 20 mins prior to formalin injection or 5 mins after the formalin injection. The formalin injection elicited an immediate and continuous discharge or burst activity in the neurons that lasted at least 90 min. Inhaled isoflurane was found to markedly inhibit this neuronal discharge. At 30 mins after discontinuing of isoflurane, the neuronal activity in both groups recovered to approximately 55% of the control value. These results suggest that formalin injection will reliably produce a continuous burst of neuronal activity in the spinal dorsal horn of cats, and that this activity can be markedly reduced by isoflurane. The ability of isoflurane to inhibit this neuronal response was not temporally related to the formalin injection. PMID- 9404127 TI - [Is laparoscopic cholecystectomy minor invasive surgery?]. AB - We conducted a comparative study to investigate whether or not laparoscopic cholecystectomy is less invasive and safer than abdominal subcostal laparotomy, using findings in blood pressure, heart rate, blood gas and endocrinological functions. In laparoscopic cholecystectomy, the patients' hospitalization period and expense for surgery were markedly reduced. However, those patients who underwent laparoscopic cholecystectomy had increases in blood pressure, heart rate, plasma cortisol and norepinephrine concentrations; it was assumed that these increases might be the result of reduced venous return to the heart which accompanied increased intra-thoracic and abdominal pressures, with reduction of cardiac output. In conclusion, since laparoscopic surgery is very advantageous for patients, we consider that for the success of this surgical procedure, it will be necessary to conduct safe anesthetic managements as well as to select patients carefully. PMID- 9404128 TI - [Postoperative nausea and vomiting after gynecologic abdominal surgery--a comparison of propofol versus inhalational technique]. AB - Propofol has been reported to reduce emesis. This study was performed to evaluate the incidence of postoperative nausea and vomiting (PONV) in gynecologic abdominal surgery patients after propofol anesthesia and inhalational anesthesia. Sixty patients were evaluated for the incidence of PONV. Thirty patients received oxygen-propofol-epidural anesthesia (propofol group) and the others were maintained with nitrous oxide-oxygen-isoflurane/sevoflurane--epidural anesthesia (inhalational group). The incidence of PONV was 33.3% in propofol group and 60% in inhalational group (P < 0.05). The means of frequency of postoperative nausea were 0.63 and 1.97 in propofol-group and inhalational group, respectively (P < 0.05). Those of postoperative vomiting were 0.17 after propofol and 1.00 following inhalational anesthesia (P < 0.01). For the gynecologic abdominal surgery patients, PONV was significantly less following intravenous anesthesia with propofol than after isoflurane or sevoflurane inhalational anesthesia. This study indicated that propofol anesthesia was useful in reducing PONV after gynecologic abdominal surgery. PMID- 9404129 TI - [Anesthesia in two patients with essential thrombocythemia]. AB - Case 1. The patient was a 69-year-old man with essential thrombocythemia (ET), who underwent urgent laparotomy. On admission he was dehydrated and the platelet count was more than 160 x 10(4).microliter-1, with hematocrit of 50%. Anesthesia was induced with ketamine i.v. and maintained with nitrous oxide and sevoflurane in oxygen. Postoperative care included the administration of gabexate mesilate (GM) which is an antiplatelet agent. Case 2. An 84-year-old woman with ET was diagnosed as gastric cancer and elective gastrectomy was scheduled. The platelet count was more than 100 x 10(4).microliter-1. The patient was anesthetized with nitrous oxide and oxygen supplemented with fentanyl and mepivacaine via epidural catheter. Intravenous infusion of GM was performed at a rate of 1 mg.kg-1.hr-1 during surgery. PF-4 and beta-TG were measured. These are platelet releasing factors. The level of PF-4 decreased to normal level during this procedure. In conclusions, it will be important to use GM during anesthesia in order to avoid the complications such as myocardial or pulmonary infarction caused by thrombocythemia. PMID- 9404130 TI - [Regional cerebral hypoperfusion reduces the effect of rectal midazolam in children with Moyamoya disease]. AB - To investigate the effect of regional cerebral blood flow on the effect of midazolam, we evaluated 99mTc-hexamethylpropylene-amine-oxime-single photon emission computed tomography (SPECT) in 37 cases of childhood moyamoya disease. They were divided into two groups according to the findings of SPECT; one group showed hypoperfusion in the bifrontal regions (n = 20), and the other did not (n = 17). Both groups received 1 mg.kg-1 of midazolam transrectally 30 min before the anesthesia induction and level of sedation was measured with six point scales. Significantly lower level of sedation score was recognized in the group that showed hypoperfusion in bifrontal regions (P < 0.05). Our finding may suggest that regional cerebral hypoperfusion may modify the sedative effect of midazolam in children with moyamoya disease. PMID- 9404131 TI - [Two cases of severe metabolic acidosis after monitoring of evoked spinal cord potential]. AB - Evoked spinal cord potential (ESCP) elicited by direct spinal cord stimulation has been applied to monitor the spinal cord function during spinal surgery. We report here two patients who showed excessive muscle contraction and developed severe metabolic acidosis as a result of the stimulation of spinal cord to measure ESCP. These changes were prevented by vecuronium administration prior to ESCP monitoring. These results suggest that excessive muscle contraction by direct spinal cord stimulation may lead to metabolic acidosis. In conclusion, the use of a non-depolarizing muscle relaxant is recommended to prevent the adverse effects of ESCP monitoring. PMID- 9404132 TI - [Anesthesia management for a patient with pulmonary eosinophilic granuloma associated with bilateral pneumothorax--general anesthesia with positive pressure ventilation]. AB - Pulmonary eosinophilic granuloma (PEG) is a rare fibroinflammatory disorder and is characterized by a mixed interstitial infiltrate of eosinophils, lymphocytes and Langerhans cells. We experienced anesthetic management for a patient with PEG associated with bilateral pneumothorax. Anesthesia was maintained with epidural anesthesia and general anesthesia using positive pressure ventilation. At first we ventilated with a low airway pressure, but recognized an increase in endtidal CO2. Then we switched to a high airway pressure (= 22 cmH2O) and decreased the concentration of carbon dioxide. No new air leaks were observed by ventilating with the high airway pressure. It was concluded that in anesthetic management for PEG we must be careful about the airway pressure during operation. PMID- 9404133 TI - [Effects of human atrial natriuretic peptide on hemodynamics and pulmonary gas exchanges in cardiac surgery patients]. AB - We investigated the effects of human atrial natriuretic peptide (hANP) on hemodynamics and pulmonary gas exchanges in 22 cardiac surgery patients without pulmonary hypertension. In 10 patients, hANP was infused at a rate of 0.2 microgram.kg-1.min-1 throughout the surgery (hANP group), while in other 12 patients hANP was not infused at all (control group). Before cardiopulmonary bypass (CPB), mean arterial pressure and systemic vascular resistance decreased and cardiac output increased significantly in hANP group as compared with those in control group. After weaning from CPB and at the completion of surgery there was no significant difference in these hemodynamic variables between the two groups. Mean pulmonary arterial pressure, pulmonary vascular resistance, arterial pH, arterial oxygen tension, arterial carbon dioxide tension and shunt ratio did not show any significant difference between the two groups throughout surgery. These findings indicate that hANP infusion causes greater systemic vasodilation with less pulmonary vasodilation, and suggest that this systemic vasodilating effect contributes to the improvement of left ventricular function in patients undergoing open heart surgery. PMID- 9404134 TI - [Pre- and post-operative management of cesarean section in a parturient with severe preeclampsia accompanied by hyperdynamic state]. AB - A 37-year-old parturient with severe preeclampsia accompanied by pulmonary edema underwent emergency cesarean section. Pulmonary artery (PA) catheter inserted while the patient was awake revealed hyperdynamic state with increased cardiac index and preload, and decreased systemic vascular resistance. Epidural anesthesia and analgesia were provided with a satisfactory outcome. Monitoring of PA pressure and cardiac index was continued postoperatively in ICU for fluid management. We conclude that preoperative PA catheterization provides useful hemodynamic information in severe preeclamptic patients associated with persistent oliguria, pulmonary edema and hyperdynamic state. PMID- 9404136 TI - [Coronary artery spasm during anesthetic induction in a patient with bronchial asthma]. AB - A 68-year-old man was scheduled for subtotal gastrectomy. He had bronchial asthma, but had no history of ischemic heart disease and showed normal ECG. He stopped taking antiasthmatic drugs after the admission. His operation had been postponed for 10 days for an attack of bronchial asthma. The asthmatic attack was suppressed by infusing aminophilline. Before the operation, general anesthesia combined with epidural anesthesia (mepivacaine; 60 mg) was induced. At the time of the insertion of a stomach tube, bradycardia (48 bpm) and hypotension (48/30 mmHg) with an elevation of ST-segment in ECG were observed. We administrated 10 mg of isosorbide dinitrate followed by continuous intravenous injection (0.5 mg.kg-1.min-1) of dopamine (6 mg.kg-1.min-1). After 20 minutes, increases of both blood pressure (82/49 mmHg) and heart rate (89 bpm) were achieved and ST-segment in ECG was reversed. The operation was postponed again. Although the patient had refused to take coronary angiogram, the episode was explained by coronary artery spasm. Pathogenesis of the spasm was likely to be 1) elevation of endogenous cathecolamine due to asthma, 2) inhibition of cardiac sympathetic system by epidural anesthesia and 3) stimulation of vagal system by the insertion of a stomach tube. PMID- 9404135 TI - [Anesthesia for a patient with macrothrombocytopenia]. AB - A 56-year-old female with macrothrombocytopenia was scheduled for colectomy and hepatectomy. She had not shown significant bleeding tendency. Her preoperative platelet counts were 0.5-1.6 x 10(4) microliters-1 with the use of an automated cell counter. However, microscopic examination showed platelet number ranging 0.9 3.4 x 10(4) microliters-1 and many macrothrombocytes. Therefore, platelet biomass (platelet number x platelet volume) seemed almost normal. Bleeding time was 3 minutes and platelet function was normal. She received preoperatively high dose gamma-globulin administration and intraoperatively platelet transfusion. The operation was performed under combined epidural and general anesthesia. Intra- and post-operative course was uneventful. PMID- 9404137 TI - [The efficacy of postural drainage in a case of pulmonary edema following cholecystectomy]. AB - An 81-year-old man underwent percutaneous transluminal gallbladder drainage. As the drain was accidentally removed six days later, he received cholecystectomy. After the operation, he developed hypotension, hypoxemia and ST level depression on ECG. He received artificial ventilation and cathecholamines. His chest CT showed marked pulmonary edema, and total protein of the edema-fluid was 3.3 g.dl 1. These findings suggested permeability pulmonary edema. He received postural drainage of the edema-fluid, and the pulmonary oxygenation was gradually improved. He was weaned from artificial ventilation on the 6th ICU day and discharged the next day. PMID- 9404138 TI - [Cuff damage during naso-tracheal intubation for general anesthesia in oral surgery]. AB - In our hospital, twenty-one cases of endotracheal tube cuff rupture during naso tracheal intubation were noted in cases using 725 polyvinyl chloride (PVC) tracheal tubes. We analysed the causes of cuff troubles in these 21 samples of tubes. When the cuffs were inflated, they were not capable of containing the air in most cases. Some cuffs had small holes (described as pinholes), and the others had longer slits on scrape marks and burst. These scrape marks may have been caused by the object with sharp edges such as spina or crista of the nasal septum, or otherwise by the tip of intubation forceps. The cuff material appeared to be slightly hardened in some samples which may be due to the lubrication. We usually lubricated the tube with lidocaine spray or gel formulation and then sometimes placed it in hot water to soften it for avoiding naso-mucosal injury. It is not generally recommended to place tubes in hot water, as this procedure may soften the cuff and make it more suspectible to damage. The clarification is also needed on the use of lidocaine. Although the gel formulation is acceptable, but the spray formulation is known to react with cuff material and make it more susceptible to inducing blistering, pinholes and sudden rupture of PVC cuffs. We conclude that these cuff damages might have occurred from various causes. A main cause must be passing the tube through the narrow nasal turbinate with spina or crista. Other causes could not only be the use of Magill forceps but also lubrication of the tube with lidocaine spray and placing it in hot water. PMID- 9404139 TI - [Anesthetic management of patients undergoing irrigation and drainage of chronic subdural hematoma--retrospective analysis of analgesia and sedation under locoregional anesthesia]. AB - We retrospectively investigated the relationship between intraoperative hemodynamic variability and variables including patient background, anesthetic profile, and operative profile in 108 patients undergoing irrigation and drainage of chronic subdural hematoma under locoregional anesthesia. Patients were divided into two groups according to the degree of changes in mean arterial blood pressure (MAP) and heart rate (HR) during operation. Group A (n = 66) had MAP and HR changes < 20%, and Group B (n = 42) had MAP or HR changes > or = 20% of preoperative baseline values. Age was significantly higher in group B than Group A (mean +/- SD; 70 +/- 12 vs. 62 +/- 14 years, P < 0.01). In respect to additional anesthetic agents used (none; no additional anesthetic agents, enough; pentazocine > or = 0.3 mg.kg-1 and droperidol > or = 0.05 mg.kg-1, pentazocine > or = 0.5 mg.kg-1 or droperidol > or = 0.15 mg.kg-1, little; less than "enough"), ratio of "little" administered in Group B was significantly higher than that in Group A (64.3% vs. 37.9%, P < 0.05). These findings suggest that intraoperative hemodynamic variability under locoregional anesthesia in patients with chronic subdural hematoma is associated with age and insufficient use of hypnotic and/or analgesic agents. PMID- 9404140 TI - [Which laryngoscope is the most stressful in laryngoscopy; Macintosh, Miller, or McCoy?]. AB - Stress responses during laryngoscopy were compared among the situations using three different laryngoscopes, Macintosh (curved standard blade), Miller (straight blade), or McCoy (levering). Blood pressure, heart rate (in 58 patients) and plasma concentration of catecholamines (in 29 patients) were measured before, during and after laryngoscopy without tracheal intubation. Systolic blood pressure after laryngoscopy was significantly higher in the Miller group than in other two groups. Plasma epinephrine concentrations after laryngoscopy in the McCoy group were lower than other two groups. Heart rate and plasma norepinephrine concentration were not different among the three groups. These results suggest that the stress response during laryngoscopy without intubation is the biggest in using the Miller laryngoscope and the smallest in using the McCoy laryngoscope. PMID- 9404141 TI - [A case of probable Creutzfeldt-Jakob disease with a point mutation of prion protein gene codon 180 and atypical MRI findings]. AB - A case of probable Creutzfeldt-Jakob disease (CJD) is reported. A 70-year-old Japanese woman was admitted with a complaint of amnesia. She initially developed Kluver-Bucy syndrome, hypergraphia, and later showed myoclonus and startle response. She developed akinetic mutism within nine months from the onset. Prion protein gene codon 180 point mutation (Met/Ile) was detected and we diagnosed her as CJD. Serial MRI studies revealed abnormal T2 elongation and thickening limited to the cerebral cortices, which started at bilateral temporal lobes and later extended to frontal, parietal, and occipital lobes with relative sparing of hippocampi and central gyri. Serial EEG did not show periodic synchronous discharge (PSD). Three cases of CJD with a point mutation of codon 180 were reported in the past. There are several common features in the past cases and the present case, i.e. 1) negative familial history, 2) late onset, 3) T2 high intensity at cerebral cortices on MRI, and 4) negative PSD. These may be characteristic features of CJD with a point mutation of codon 180. PMID- 9404142 TI - [Two cases of pure akinesia with unusual activation in dorsal part of the frontal lobe during gait--surface EMG and PET study]. AB - FDG-PET activation study was performed in two pure akinesia (PA) patients with silent pattern in surface electromyography (EMG) frozen gait analysis. Three patients (2 PA and 1 PD with reciprocal co-contraction pattern) and 7 normal subjects underwent FDG-PET study in two different condition of "rest" and "walk", in order to evaluate the change in regional cerebral activity during gait. In a PD patient, the activated pattern was no different from that of normal individuals; the cerebellar vermis was solely activated significantly. In contrast, 2 PA patients showed characteristic activation in the dorsolateral frontal area in addition to the areas found in normals. These results suggest that the dorsolateral frontal cortex may play an inhibiting role to the motor cortex resulting in the silent pattern of frozen gait in PA. Combined study of surface EMG and FDG-PET during gait may be useful for analyzing the pathophysiological mechanism of gait disturbance. PMID- 9404144 TI - [A case of malignant rheumatoid arthritis with transverse myelopathy and multiple lacunar infarction]. AB - We reported a 34-year-old woman with malignant rheumatoid arthritis (MRA) associated with transverse myelopathy and multiple lacunar infarction. She had suffered from MRA for 9 years, then developed sensory disturbance of left big toe and weakness of right lower limb. Neurological examination revealed the muscle weakness of right lower limb. Deep tendon reflexes were hyperactive in bilateral lower limbs. Babinski's sign was positive bilaterally. Superficial sensation was decreased below Th10 level on the left side. Urinary bladder and rectal disturbance were not present. Laboratory examination disclosed perinuclear antineutrophil cytoplasmic autoantibody (p-ANCA) and anti-nuclear antibody (ANA). Multiple lacunar infarction and syringomyelia were found by MRI studies. Histological examinations of skin ulcer biopsied at 17 years of age disclosed vasculitis. We speculated that vasculitis associated with MRA might cause the damage of central nervous system (CNS) in our case. The p-ANCA may accelerate the vasculitic changes in CNS. PMID- 9404143 TI - [A case of incomplete Kearns-Sayre syndrome with a stroke like episode]. AB - A 32-year-old woman developed chronic progressive hearing impairment, trunkal ataxia, bilateral ptosis and external ophthalmoplegia. She also showed slowly progressive mild to moderate proximal dominant muscle weakness and atrophy. ECG showed incomplete right bundle branch block. An aerobic exercise test showed abnormal blood lactate elevation and muscle biopsy revealed ragged-red fibers in addition to the myopathic change. Analysis of mitochondrial DNA extracted from biopsied muscle and fibroblast samples revealed a 1,758bp deletion from the cytochrome b to ND6 coding regions. Common mutations in tRNALeu(UUR) coding region to the mitochondrial encephalomyopathy, lactic acidosis and stroke-like episodes (MELAS) were not present. She was diagnosed as having incomplete Kearns Sayre syndrome (KSS). Since the age of 35, she developed complex partial seizure attacks with secondary generalization frequently and at the age of 42, she had a severe generalized seizure with delayed consciousness loss followed by left hemiplegia. MRI showed wide T2-high signal lesions in the right temporo-parieto occipital area. The proton MR-spectroscopy showed prominent increase of lactate beyond the lesions detected by MRI, indicating diffuse aerobic metabolic dysfunction in the central nervous system. We reviewed two other KSS cases with a stroke like episode, who also had epilepsy and large deletion but no tRNALeu(UUR) mutation, in mitochondrial DNA. Patients with KSS who have seizure may develop the stroke-like episode as seen in MELAS patients. PMID- 9404145 TI - [A family with facioscapulohumeral muscular dystrophy and hereditary long QT syndrome]. AB - We describe a family with facioscapulohumeral muscular dystrophy (FSHD) and hereditary long QT syndrome (LQT) for three generations. The proband, a 50-year old woman, had noticed difficulty in raising the upper extremities since the age of 40. At the age of 48, she was admitted to our hospital because of arrhythmia attack. She, her mother, and one of her three children were diagnosed as having LQT. These three individuals and the proband's two siblings were clinically diagnosed as having FSHD which was confirmed by genetic analysis using EcoR1. FSHD is an autosomal dominant disorder and the gene locus is mapped to chromosome 4q35-ter, but the gene has not been isolated. LQT is a group of disorders which cause syncope and sudden death from ventricular arrhythmia in an autosomal dominant fashion. Four loci for this syndrome (LQT1-4) have been known, and three of the genes have been shown to encode ion-channels. Genetic analysis of the proband failed to detect any of previously known mutations in the LQT1, LQT2, and LQT3 genes. The locus for LQT4 has been mapped to chromosome 4q25-7. There have so far been no report of FSHD associated with LQT. Although the pathogenesis is unclear, we speculate that these two diseases are linked each other on chromosome 4q. PMID- 9404146 TI - [An adult case of bacterial meningitis caused by penicillin-resistant Streptococcus pneumoniae with gene mutations of penicillin binding proteins]. AB - A 62-year-old Japanese male developed acute bacterial meningitis. Administration of ABPC, CDZM, INH, and RFP was started. The causative bacteria was identified as penicillin-resistant Streptococcus pneumoniae (PRSP) 3 days later. We changed ABPC to PAPM/BP and the bacterial meningitis improved dramatically. Measurement of minimum inhibitory concentration (MIC) of the bacterial strain from CSF and blood showed that it is susceptible to PAPM/BP and RFP. Mechanism of drug resistance is considered to be gene mutations of penicillin binding protein (PBP) that decreases drug affinity to PBP. Recently penicillin resistance of Streptococcus pneumoniae has been found to be associated with mutations of pbp 2b, 1a genes and the cefem resistance of that is with those of pbp-2x, and 1a genes. By PCR (polymerase chain reaction) analysis we identified the mutations of pbp-1a, 2x, and 2b genes of the isolated strain of Streptococcus pneumoniae. We emphasize the importance of detection of pbp gene mutations for the diagnosis of PRSP infections. PMID- 9404147 TI - [Hyperglycemia and symmetrical proximal neuropathy in diabetes]. AB - A 60-year-old woman, diagnosed as having a diabetic symmetrical proximal motor neuropathy, is presented. In March 1995, she was referred to us because of untreated diabetes mellitus since 1990. Insulin treatment during one month decreased her postprandial plasma glucose level from more than 400mg/dl to about 200mg/dl. Soon after the treatment, she noticed lower proximal limb weakness bilaterally. In several months, her weakness progressed as the fasting plasma glucose level was increased. Her muscle power gradually recovered when the plasma glucose level was normalized. We therefore suggest that metabolic changes related to hyperglycemia, rather than ischemic vascular changes, play an important role in the pathogenesis of a diabetic symmetrical proximal motor neuropathy. PMID- 9404148 TI - [A case of polyneuropathy with B-CLL and HTLV-I associated myelopathy (HAM)]. AB - We report a case of polyneuropathy associated with chronic B lymphoid leukemia (B CLL) and HAM. A 65-year-old man had an initial onset with polyneuropathy, then myelopathy appeared. High HTLV-I antibody and anti-ganglioside antibodies were observed. The biopsy examination of the sural nerve showed mainly demyelinated changes. Muscle weakness gradually improved after cyclophosphamide therapy. Anti ganglioside antibodies were also decreased in titer. This case suggested that the production of antiganglioside antibodies and the occurrence of polyneuropathy might be caused by both factors of HTLV-I infected T cells and B cell tumors. PMID- 9404149 TI - [A case of Kallmann syndrome with empty sella and arachnoid cyst]. AB - We report a 28-year-old man of Kallmann syndrome with arachnoid cyst and empty sella. At age 22, he was admitted with acute slipped capital epiphysis and diagnosed as primary hypogonadotropinemia, because of no response to LH-RH before and after 7-day LH-RH injection. He was treated with androgen for only one year. On his second admission due to femoral head necrosis at age 28, the endocrinological evaluation suggested hypothalamic hypogonadotropinemia. Although he had mild hyposomia, we diagnosed him as Kallmann syndrome, because abnormalities of rhinencephalon was present on MRI. Arachnoid cyst in the middle cranial fossa and empty sella were also observed on MRI and the ballooning of the sella had been advanced on plain X-ray for these 6 years. As Kallmann syndrome is known to be accompanied with midline craniofacial anomalies, the dysplasia of sellar diaphragm might be originated by the same pathogenesis. In this case, empty sella might be caused by impaired CSF dynamics due to arachnoid cyst as well as possible constitutional anomaly of the diaphragm. PMID- 9404150 TI - [An apparently sporadic case with spinocerebellar ataxia type 1 (SCA1)]. AB - We reported a sporadic case with late onset SCA1. There was no family history of neurological diseases. His parents had been healthy until they died at the age of 77 and 89 years, respectively. The patient noticed gait disturbance at age of 60. Thereafter, he gradually developed cerebellar ataxia, hyporeflexia, mild atrophy of the facial and limb muscles and moderate deep sensory disturbance. MRI of the brain showed moderate atrophy of the cerebellum and brainstem. Sequencing analysis of SCA1 gene demonstrated that the patient had an expanded allele with 40 CAG repeats and no CAT interruption. Consequently, he was diagnosed as having SCA1. These results suggest the possibility that among apparently sporadic cases with cerebellar ataxia, there are some cases of SCA1 with mild CAG repeat expansion. PMID- 9404151 TI - [A case of lumbosacral lipoma-associated adult onset tethered cord syndrome with initial symptoms of sensory disturbance and intractable foot ulcers]. AB - A 63-year-woman who complained of sensorimotor disturbance of the lower extremities and urinary disturbance was presented. She noted loss of superficial sensation in both feet and foot ulcers at the age of 20 years. Her illness was initially diagnosed as hereditary sensory neuropathy type 1 (HSN1). The foot ulcers were so intractable that she had to have her right leg amputated at the age of 48 years. She had a severely impaired superficial sensation in the lower extremities and buttock, distal weakness of the left leg, and dysuria at the age of 60 years. The neurological examination revealed that she had segmental sensorimotor disturbance below the levels of the 5th lumbar segment. MRI demonstrated tethered cord with a lumbosacral lipoma. Adult onset tethered cord syndrome (TCS) that presents with HSN 1-like symptoms as initial clinical features has not yet been reported. Foot ulcers are often seen in child onset TCS in which the degree of tethered cord is severer than adult onset cases. It is reported that release of the tethered cord promotes healing of the foot ulcers. We recommend MRI for the study of the lumbosacral cord of patients with HSN 1 like symptoms, because there is a possibility that such patients may have TCS and early surgical treatment is effective for TCS. PMID- 9404152 TI - [Cataract as an only clinical manifestation of myotonic dystrophy--a new example of somatic instability of CTG repeats expansion in myotonin protein kinase gene]. AB - We reported a 69-year-old woman who developed a cataract as a single clinical expression of myotonic dystrophy (MD). There are many MD patients in her family including her 29-year-old daughter suffering from congenital MD. We compared CTG repeats expansion in the motonin protein kinase gene from the lens obtained at operation with that of her and her daughter's lymphocytes using two techniques, Southern blot and PCR amplification analyses. On the Southern analysis using Eco RI and BGII-digested DNA, lymphocytes from the patient and her daughter showed a band of abnormal size with about 80 and 2,500 repeats respectively in addition to the normal size allele. No band was visualized in the patient's lens because the amount of DNA was presumably too small. On PCR analysis the band of expanded allele was observed in the patient's lymphocytes only whereas the band of normal allele was present in all the specimens. The expanded CTG repeats were too large to be amplified by PCR as demonstrated on the Southern blot of her daughter's lymphocyte. Consequently the discrepancy in the size of CTG repeat expansion in lens and lymphocyte is a new example of somatic instability of the repeat which explains the cataract as an only clinical manifestation of MD in this patient. PMID- 9404153 TI - [A case of multiple cerebral arterial thrombosis due to congenital protein C deficiency]. AB - We report a 49-year-old man who had right hemiparesis and motor aphasia. A computed tomography revealed hypodense areas in the left frontal subcortex. A cerebral angiography demonstrated occlusion of the left distal internal carotid artery and both anterior cerebral arteries, as well as stenosis of the left internal carotid artery at the cervical portion. The second angiogram obtained a month later showed no changes. The diagnosis of atherothrombotic cerebral infarction was established on the basis of clinical profile and angiographic findings. Protein C activity and antigen levels were reduced to approximately one half of the normal level in the patient and his brother. The patient had no other risk factors for stroke. Protein C deficiency has been considered one of the risk factors for thrombotic diseases. Venous thrombosis is the most common clinical manifestation, whereas arterial thrombosis is relatively rare. It is generally believed that arterial ischemic stroke associated with protein C deficiency occurs with embolic mechanism, and atherothrombotic infarction is extremely rare. This is the first report suggesting the possibility that protein C deficiency can cause cerebral thrombosis. PMID- 9404154 TI - [Cell cycle switch from mitotic to meiotic in fission yeast: critical role for an RNA-binding protein]. PMID- 9404155 TI - [Non-mevalonate pathway. A new pathway for the biosynthesis of isopentenyl diphosphate]. PMID- 9404156 TI - [Glycolipids in chloroplast thylakoid membrane: the mechanism for the biosynthesis and evolutionary origin]. PMID- 9404157 TI - [Phycobilisomes: supramolecular assembly in cyanobacteria for capturing of light energy]. PMID- 9404158 TI - [Genomic imprinting and its related diseases]. PMID- 9404159 TI - [Glutamate production of coryneform bacteria]. PMID- 9404160 TI - [Gene modified foods and their safety]. PMID- 9404161 TI - Multiple trauma and massive transfusion. PMID- 9404162 TI - Auditory evoked potential index: a quantitative measure of changes in auditory evoked potentials during general anaesthesia. AB - We describe a novel index derived from the auditory evoked potential, the auditory evoked potential index, and we compare it with latencies and amplitudes related to clinical signs of consciousness and unconsciousness. Eleven patients, scheduled for total knee replacement under spinal anaesthesia, completed the study. The initial mean (SD) value of the auditory evoked potential index was 72.5 (11.2). During the first period of unconsciousness it decreased to 39.6 (6.9) and returned to 66.8 (12.5) when patients regained consciousness. Thereafter, similar values were obtained whenever patients lost and regained consciousness. Latencies and amplitudes changed in a similar fashion. From all parameters studied, Na latencies had the greatest overlap between successive awake and asleep states. The auditory evoked potential index and Nb latencies had no overlap. The consistent changes demonstrated suggest that the auditory evoked potential index could be used as a reliable indicator of potential awareness during propofol anaesthesia instead of latencies and amplitudes. PMID- 9404163 TI - Staff stress on the intensive care unit: a comparison of doctors and nurses. AB - Working on an intensive care unit is perceived as stressful. This study investigated occupational stress in staff working on an intensive care unit using the occupational stress indicator. Questionnaires were given to all intensive care staff; the replies were then analysed and compared with normative data. The response rate was 62%. Intensive care unit staff found aspects of their job relating to career and achievement and organisational design and structure more stressful than a normal working population. Their coping strategies differ but the only significantly different measure of adverse outcome was related to personal relationships at work. The job itself was not found to be a significant source of stress. Nursing staff have different sources of stress from medical staff and individuals with partners or children are relatively protected from stress. PMID- 9404165 TI - Effects of cardiopulmonary bypass and hypothermia on electroencephalographic variables. AB - We studied the effects of hypothermia and cardiopulmonary bypass (CPB) on four depth of anaesthesia monitors; spectral edge frequency (SEF), median frequency (MF), bispectral index (BIS) and auditory evoked potential index (AEPIndex) in 12 patients during uneventful cardiac anaesthesia. Each variable was recorded simultaneously at 10 periods during anaesthesia. All four variables were not affected by the transition to CPB. During hypothermia, values of AEPIndex, MF and SEF were tightly distributed but values of BIS were very variable and overlapped with those before induction of anaesthesia. The variability decreased during rewarming. The values of AEPIndex throughout the anaesthesia never overlapped with those before induction of anaesthesia. The AEPIndex was the most stable and reliable as a depth of anaesthesia monitor among the four variables in cardiac bypass surgery. PMID- 9404164 TI - An audit of the safety of an acute pain service. AB - We audited and analysed the adverse effects and safety of postoperative pain management on 2509 consecutive patients under care of the Acute Pain Service at a tertiary referral teaching hospital over a 32-month period. Our standard respiratory monitoring consisted of continuous pulse oximetry, hourly respiratory rate counting, sedation scoring and intermittent arterial blood gas sampling. This protocol was reliable and detected six episodes of bradypnoea, 13 of hypercapnia and 23 of oxygen desaturation occurring in 39 patients (1.8% of all spontaneously breathing patients). Two patients required naloxone injection and none had long-term sequelae. Hypotension due to epidural bupivacaine 0.0625% and fentanyl 3.3 micrograms.ml-1 infusion occurred in four patients (1.2%), all with a sensory block higher than T5. They readily responded to fluid infusion and ephedrine (two patients). Postoperative nausea or vomiting occurred in 723 (28.8%) and 380 (15.1%) patients, respectively. Odds ratio analysis showed that the risk factors for postoperative nausea and vomiting were: female gender, gynaecological operations, nongeriatric patients and systemic analgesia. Postoperative nausea and vomiting decreased analgesic efficacy by discouraging the use of patient-controlled analgesia and was regarded as equally distressing as pain. Other side-effects included: pruritus in 182 patients; dizziness in 333 and lower limb weakness in 73 (21.2% of patients receiving epidural local anaesthetics). It is concluded that a standard monitoring and management protocol, an experienced nursing team and reliable Acute Pain Service coverage is mandatory for the safe use of modern analgesic techniques. PMID- 9404166 TI - Minimum alveolar concentration for halothane in children with cerebral palsy and severe mental retardation. AB - Children with cerebral palsy and severe mental retardation who present for operation may require lower concentrations of inhalational anaesthetics than healthy children. The minimum alveolar concentration (MAC) for halothane was measured in 36 children and adolescents, aged 4-18 years, who underwent orthopaedic surgery. The control group consisted of 12 healthy children (group 1). Children with cerebral palsy and severe mental retardation were allocated to one of two groups: those taking chronic anticonvulsant medication (group 2) (n = 12) and those who did not take any drugs (group 3) (n = 12). The mean (SEM) MAC value for halothane (expressed in volume per cent) was 0.90 (0.02) for healthy children. Children with cerebral palsy had significantly lower MAC values whether they took anticonvulsant drugs or not (0.62 (0.03) and 0.71 (0.10), respectively). PMID- 9404167 TI - In vitro evaluation of the effect of profound haemodilution with hydroxyethyl starch 6%, modified fluid gelatin 4% and dextran 40 10% on coagulation profile measured by thromboelastography. AB - Synthetic colloids have been implicated as a cause of coagulopathy when administered in large quantities. The effect of profound haemodilution (50%) on coagulation profile was measured in vitro by thromboelastography. Blood samples were taken from 11 ASA grade 1 patients prior to induction of anaesthesia for elective surgery. Each sample was simultaneously tested in four different preparations: undiluted blood (control sample); blood diluted with hydroxyethyl starch 6%; blood diluted with modified fluid gelatin 4%; blood diluted with dextran 40 10%. There was a significant decrease in reaction time in the preparations treated with hydroxyethyl starch 6% and modified fluid gelatin 4%, reflecting activation of initial fibrin formation. A significant increase in clot formation time was noted in the hydroxyethyl starch 6%-treated preparations. There was also a significant decrease in clot formation rate and maximum amplitude in the hydroxyethyl starch 6% group. Clot formation time, clot formation rate and maximum amplitude did not change in the modified fluid gelatin 4% group. Profound haemodilution with dextran 40 10% exerted extreme effects on the measured variables, very often resulting in a straight line on the thromboelastography profile. PMID- 9404168 TI - Serum electrolyte and blood gas changes after intrathecal and intravenous bolus injections of magnesium sulphate. An experimental study in a rat model. AB - The effect of intrathecally administered magnesium sulphate on serum levels of magnesium, sodium, potassium, calcium and blood gas variables was studied in a rat model. Magnesium sulphate given intrathecally has previously been shown to produce segmental spinal blockade with no permanent neurological damage. The previous studies, however, had not investigated the possible systemic effects of the magnesium sulphate. The serum magnesium level increased significantly at 1 and 2 h after the intrathecal injection of both 6.3% and 12.6% magnesium sulphate (6.3%: 28% at 1 h, 24% at 2 h; 12.6%: 22% at 1 h, 16% at 2 h). These changes were not as great as occurred when the same dose of magnesium sulphate was administered intravenously. In all cases, the serum magnesium had returned to normal by 24 h. There were no significant changes in calcium, sodium or potassium levels, nor in arterial blood gas variables. These results show that intrathecally administered magnesium sulphate has little effect on electrolyte homeostasis. PMID- 9404169 TI - An inexpensive, self-assembly pressure algometer. AB - We have developed a simple instrument for pressure algometry. It can be made easily using components found in most anaesthetic rooms. Ten students were able to make the device using written instructions. All the resulting algometers performed within 10% accuracy limits for values up to 4 kg.cm-2. PMID- 9404170 TI - Convection warmers--not just hot air. AB - We sought to determine whether the forced air convection warmers (nine Bair Huggers, Augustine Medical, and one Warm Touch, Mallinkrodt Medical) used in our operating theatres could be a source of microbial pathogens. Agar plates were placed directly in the air stream of the warmers. Four of these grew potentially pathogenic organisms. When the warmers were set to blow through perforated blankets, no growth occurred. Three of the warmers were swabbed and sites of colonisation were found in their hoses. After fixing a microbial filter to the end of the hose, organisms were no longer detectable. We conclude that these warming devices are a potential source of nosocomial infection. They should only be used in conjunction with perforated blankets, should have their microbial filters changed regularly and their hoses sterilised. The inclusion of a microbial filter into the nozzle of the hose could be incorporated into the design of the warmer. PMID- 9404171 TI - Anaesthetic pollution. Potential sources, their identification and control. AB - Anaesthetic gases escape into the ambient air mainly from the anaesthetic breathing system but there are many other important sources of anaesthetic pollution. These include the filling of vaporisers, inhalational induction techniques, leaks around the patient's face mask, leaks from monitoring equipment and loose-fitting or perishing equipment. The control of substances hazardous to health (COSHH) regulations together with the recent implementation of the occupational exposure standards (OES) for anaesthetic gases require that any risk to health workers from anaesthetic exposure be assessed, control measures implemented, the environment monitored and OES not exceeded. The installation of costly scavenging equipment is believed to have reduced the levels of pollution in operating theatres, but several independent reports suggest that their use has not been wholly effective. Many sources of pollution remote from scavenging equipment may be responsible for the ineffective control of anaesthetic pollution. During an initial COSHH assessment in our hospital, using an infra-red analyser specifically designed for leak testing and background monitoring, we have identified several controllable sources of pollution. These included leaks from an ill-fitting face mask, loose connections in the anaesthetic breathing system, leaks from the laryngeal mask at the end of the operation and a leak of more than 5400 ppm N2O was found near the unscavenged gas outlet at the back of the multigas monitor. Leak testing, using infra-red analysers with their rapid response, has been recommended as an important aid in the identification of hidden sources of pollution, most of which we believe are amenable to control and remedy. PMID- 9404172 TI - Cervical osteomyelitis following tonsillectomy. AB - We present a case of fatal cervical osteomyelitis following an elective tonsillectomy in a previously fit young man. Following induction of general anaesthesia, and prior to surgery, the patient received bilateral glossopharyngeal nerve blocks with 0.5% bupivacaine and adrenaline 1:200,000. The initial recovery was uneventful but persistent throat and neck pain developed at home which was diagnosed as a throat infection and possible hyperextension injury of the neck. It is impossible to say how much the dissection of chronically infected tonsils or the infiltration of local anaesthetic into or near a potentially infected area contributed to the development of cervical osteomyelitis. The absence of any other symptoms and signs, a normal blood count and cervical spine X-ray, and the rarity of cervical osteomyelitis, all contributed to a delay in diagnosis. PMID- 9404173 TI - Management of severe tracheal obstruction with helium/oxygen and a laryngeal mask airway. PMID- 9404174 TI - Thoracic epidural anaesthesia for coronary artery bypass graft surgery. Effects on postoperative complications. AB - We have performed a retrospective analysis of the peri-operative course of 218 consecutive patients who underwent routine coronary artery bypass graft surgery in this institution. All patients received a standardised general anaesthetic using target-controlled infusions of alfentanil and propofol. One hundred patients also received thoracic epidural anaesthesia with bupivacaine and clonidine, started before surgery and continued for 5 days after surgery. The remaining 118 patients received target-controlled infusion of alfentanil for analgesia for the first 24 h after surgery, followed by intravenous patient controlled morphine analgesia for a further 48 h. Using computerised patient medical records, we analysed the frequency of respiratory, neurological, renal, gastrointestinal, haematological and cardiovascular complications in these two groups. New arrhythmias requiring treatment occurred in 18% of the thoracic epidural anaesthesia group of patients compared with 32% of the general anaesthesia group (p = 0.02). There was also a trend towards a reduced incidence of respiratory complications in the thoracic epidural anaesthesia group. The time to tracheal extubation was decreased in the epidural group, with the tracheas of 21% of the patients being extubated immediately after surgery compared with 2% in the general anaesthesia group (p < 0.001). There were no serious neurological problems resulting from the use of thoracic epidural analgesia. PMID- 9404175 TI - Comparison of nalbuphine and buprenorphine in total intravenous anaesthesia. AB - Nalbuphine (0.3 mg.kg-1) and buprenorphine (2.5 micrograms.kg-1) were compared as part of a total intravenous anaesthesia regimen using a propofol infusion in 60 patients undergoing laparoscopic cholecystectomy in a randomised double-blind study. Changes in haemodynamic variables greater than 20% from the baseline were noted. No difference was observed in blood pressure but the heart rate was significantly lower in the buprenorphine group. Intra-operative bradycardia (heart rate < 60 beat.min-1) occurred more often in the buprenorphine group. Recovery was fast and comparable with both drugs and no patient reported awareness. Quality of analgesia was similar in both groups. Both drugs provide suitable analgesic supplementation to total intravenous anaesthesia. PMID- 9404176 TI - Analgesic and respiratory effect of nalbuphine and pethidine for adenotonsillectomy in children with obstructive sleep disorder. AB - Opioids may depress respiration and contribute to airway obstruction after adenotonsillectomy for obstructive sleep disorder. We compared the respiratory and analgesic effects of nalbuphine, which has a ceiling effect for respiratory depression, and pethidine in 90 children (aged 2-12 years) with a history of obstructive sleep disorder undergoing adenotonsillectomy. Children were scored for their obstructive sleep disorder history and were randomly allocated to receive intravenously at induction of anaesthesia either nalbuphine 0.1 mg.kg-1 (group N) or pethidine 1 mg.kg-1 (group P). End-tidal carbon dioxide was measured in the recovery period using a nasopharyngeal catheter and oxygen saturation whilst breathing air; pain and sedation scores were recorded for 6 h postoperatively. Both groups were similar with respect to the demographic data and respiratory measurements: mean (SD) oxygen saturation on air in the recovery area (96.2% (1.2) vs. 96.5% (1.1) in group N and P, respectively) and mean (SD) end-tidal carbon dioxide (46.4 (5.5) mmHg vs. 47.7 (4) mmHg in group N and P, respectively). High obstructive sleep disorder score, history of apnoea, hyperactivity and loud snoring were found to be the best predictors of early postoperative oxygen desaturation in both groups. PMID- 9404177 TI - The site of airway irritation during induction of anaesthesia. AB - The aim of this investigation was to study the role of the nasal airway in mediating upper airway reflexes during induction of anaesthesia when the commonly used irritant inhalational anaesthetic agent enflurane is used. In a prospective randomised study, 40 ASA 1 & 2 day-case patients undergoing body surface surgery were recruited. Following intravenous induction using propofol, 20 patients received enflurane administered via a laryngeal mask airway (LMA), the anaesthetic vapour therefore bypassing the nasal airway. In the other group, 20 patients received enflurane anaesthesia administered using a face mask, the nasal airway therefore being exposed to inhalation anaesthetic. We were unable to demonstrate any significant (p < 0.05) differences between the two groups in relation to upper airway complications (cough, breath holding, laryngeal spasm, bronchospasm and excitement). Previous work has identified the nose as a possible important reflexogenic site for upper airway reflexes in humans during anaesthesia. We have been unable to demonstrate any difference in upper airway complications when the nasal airway was included or excluded from exposure to irritant anaesthetic vapours, when administered in a clinical setting. PMID- 9404179 TI - Guidelines and cardiac anaesthetists. PMID- 9404180 TI - Cancellation of elective abdominal aortic aneurysms due to lack of ICU beds. PMID- 9404178 TI - Comparison of caudal block using bupivacaine and ketamine with ilioinguinal nerve block for orchidopexy in children. AB - Forty boys weighing less than 25 kg undergoing unilateral orchidopexy were randomly allocated to receive one of two analgesic regimens. Group C received a caudal epidural block with 0.25% bupivacaine 1 ml.kg-1 and preservative-free ketamine 0.5 mg.kg-1; Group L received an ilioinguinal nerve block with 0.25% bupivacaine 0.5 ml.kg-1 and infiltration of the wound with 0.25% bupivacaine 0.5 ml.kg-1. All subjects received diclofenac sodium 1-2 mg.kg-1 as a rectal suppository. Postoperative pain was assessed by means of a modified Objective Pain Score and analgesia was administered if this exceeded a value of 4. The median duration of analgesia was 10 h (range 2.6 to > 24 h) in Group C and 2.9 h (range 0.7 to > 24 h) in Group L (p < 0.05). There were no differences between groups in the incidence of motor block, urinary retention, postoperative vomiting or postoperative sedation. Subjects in Group L required significantly more doses of postoperative analgesia than those in Group C (p < 0.05). PMID- 9404181 TI - Direct measurement of pulsus paradoxus in acute severe asthma. PMID- 9404182 TI - Pain relief in familial Mediterranean fever. PMID- 9404183 TI - Correct choice of anti-emetic. PMID- 9404184 TI - Inadvertent dural tap related to problems with a loss of resistance device. PMID- 9404185 TI - Epidural catheter 'gels': a sticky problem. PMID- 9404186 TI - Does esmolol block the cardiovascular response to intubation? PMID- 9404187 TI - Extravasation associated with a multilumen central catheter. PMID- 9404188 TI - Sumatriptan and postdural puncture headache. PMID- 9404189 TI - Dangers of scavenging systems. PMID- 9404190 TI - Peri-operative use of nonsteroidal anti-inflammatory drugs in children. PMID- 9404191 TI - Complete airway obstruction during awake fibreoptic intubation. PMID- 9404192 TI - Predictors of smoking cessation at 6 months after surgery. PMID- 9404194 TI - A non-rebreathing coaxial anaesthesia system. PMID- 9404193 TI - Inadvertent intra-arterial propofol. PMID- 9404196 TI - Anaesthesia, anaesthetics and anaesthesiology. PMID- 9404195 TI - Oral temazepam. PMID- 9404197 TI - Can careful ultrasound examination of the urinary tract exclude vesicoureteric reflux in the neonate? AB - The aim of the study was to determine whether a urinary tract appearing normal when assessed by meticulous ultrasound (US) examination may coexist with vesicoureteric reflux (VUR) and whether a normal US scan can be used to exclude VUR, thereby avoiding unnecessary voiding cystourethrography (VCUG). The US features of 35 neonates with known VUR were reviewed. Criteria studied included pelvic dilatation above 7 mm on a transverse scan, calyceal or ureteral dilatation, pelvic or ureteral wall thickening, absence of the corticomedullary differentiation (CMD) and signs of renal dysplasia (small kidney, thinned or hyperechoic cortex and cortical cysts); all signs that have been shown to result from or to be associated with VUR. 57 refluxing renal units (RRU) were found among the 35 patients. VUR was bilateral in 22. Among the 57 RRU, at least one US anomaly that would have prompted VCUG was present in 50 (87.7%). Pelvic dilatation above 7 mm was present in 29 RRU (50.9%) only. Calyceal dilatation was present in 24 RRU, the dilatation involving the calyces but not the renal pelvis in seven. Ureteral dilatation was observed in 15 RRU. Pelvic or ureteral wall thickening was present in seven RRU. CMD was absent in 32 RRU (56.1%). US signs of dysplasia were found in 19 RRU. No US anomaly was found in seven RRU (12.3%) in six patients. A careful and meticulous US examination of the neonatal urinary tract allows the detection of over 87% of RRU by showing at least one sonographic abnormality. It is concluded that a normal appearing urinary tract on US does not usually coexist with VUR and that in such cases VCUG is not necessary. PMID- 9404198 TI - The clinical significance of fetal right heart enlargement with a structurally normal heart. AB - Isolated right heart enlargement of the fetus without cardiac structural abnormalities appears to be associated with growth retardation or abnormal perinatal outcome. We report the outcome of eight fetuses with subjective enlargement of the right heart. All cases were diagnosed antenatally from a standard four-chamber view on an ultrasound scan. All fetuses had otherwise normal cardiac anatomy. Seven out of the eight fetuses ultimately had outcomes which deviated from the norm. An isolated finding of fetal right heart enlargement warrants close antenatal fetal surveillance. PMID- 9404199 TI - The changing face of emphysematous cholecystitis. AB - Emphysematous cholecystitis is a variant of acute cholecystitis characterized by the presence of gas in the gall bladder lumen, wall or pericholecystic tissues in the absence of an abnormal communication between the biliary system and the gastrointestinal tract. In the past, the diagnosis has relied on the plain abdominal radiograph (AXR), since there are no clinical features to separate this condition from simple acute cholecystitis. The apparently high mortality and morbidity associated with emphysematous cholecystitis has previously emphasized the importance of emergency cholecystectomy. We have reviewed eight cases of emphysematous cholecystitis presenting to this hospital over the last 5 years. The diagnosis was made on AXR in only one of these cases. Ultrasound (US) scans were performed in all eight cases, of which five were positive and three negative, due to non-visualization of the gall bladder. In the three negative cases, the diagnosis was made on subsequent CT scans. On initial clinical examination, only one of the eight patients appeared systemically unwell and conservative management was employed in five of the patients. The remaining three patients underwent cholecystectomy within 3-5 days because of continuing signs or symptoms. It is concluded that the AXR is relatively insensitive in the diagnosis of emphysematous cholecystitis. As a result of the regular use of US in suspected hepatobiliary disease, emphysematous cholecystitis is being diagnosed with increased frequency, uncovering a broad spectrum of disease ranging from mild to severe. Previously, failure to separate milder cases from simple acute cholecystitis may have been responsible for reports of unremitting severity and progression requiring emergency cholecystectomy. Based on clinical assessment, conservative surgical management is possible in a significant proportion of patients. PMID- 9404200 TI - Ultrasound assessment of swallowing in malnourished disabled children. AB - Oral motor dysfunction is common in children with neurological impairment. Nutritional advice depends upon an accurate assessment of feeding potential in these cases. Videofluoroscopic assessment of oral motor function has been the accepted "gold standard" investigation for several years but has significant drawbacks, including the time constraints set by the use of ionizing radiation and the problems posed by the cumbersome equipment needed in mimicking the child's normal feeding situation. Ultrasonography (US) has been suggested as an alternative or additional investigation of oral motor function in children with neurological impairment. We prospectively evaluated a scoring system derived from US assessment of oral motor function in 32 malnourished disabled children with feeding problems by comparing them with a group of matched control children without neurological impairment. US imaging provided useful information with regard to the oral cavity and the soft tissue structures, capturing the salient features of tongue/hyoid/palate activity and bolus transport across the tongue and through the hypopharyngeal area. The mean percentage score obtained by US assessment of oral motor function in children with neurological impairment was 54.3 +/- 23.2 and from children without neurological impairment 91.9 +/- 12.7 (p < 0.0001). Scores for the oral and pharyngeal phases of swallowing were also very significantly lower than that in the control group, both phases being equally impaired in the disabled children. This study has demonstrated that a scoring system based on US assessment of different components of oral motor activity detects statistically significant differences in the feeding capabilities of children with neurological impairment. PMID- 9404201 TI - Sensitivity of computed tomography in detecting local recurrence of prostatic carcinoma following radical prostatectomy. AB - The aim of this study was to evaluate CT imaging in the post-operative follow-up and in the detection of recurrence after radical prostatectomy in cases of prostatic carcinoma. In over 500 patients undergoing radical prostatectomy for prostatic carcinoma, 22 cases with local recurrence were found. CT examinations of the pelvis were retrospectively evaluated in these patients. Local recurrence was detected by PSA uptake and confirmed by transrectal ultrasound (TRUS) in combination with guided biopsy. In 22 cases of confirmed local recurrence, positive results on CT were found in eight patients (36%) and negative results in nine patients (41%). In the remaining five cases (23%), no distinction could be made between scar and local recurrence. All cases definitively classified as recurrent tumour disease showed a soft tissue mass of 2 cm or more. CT sensitivity in local recurrence of prostatic carcinoma after surgery is low. Even in a very careful follow-up, the understaging would be up to 41%. In comparison, PSA, TRUS and needle biopsy are the methods of choice and are superior to CT imaging. Based on these results, there would be no reason for including pelvic CT examinations in the follow-up of prostatic carcinoma after radical prostatectomy. PMID- 9404202 TI - Comparison of two oral evacuants (Citromag and Golytely) for bowel preparation before barium enema. AB - Oral regimens for bowel preparation before barium enema examination are routinely used because of their convenience and simplicity, rather than the traditional method of colonic wash-out. We performed a prospective study comparing the side effects and efficacy of two commonly used oral bowel evacuants (Citromag and Golytely) for bowel preparation before barium enema examination in 102 patients. The side-effects associated with the agents were assessed by analysing a questionnaire completed by the patients. There was no significant difference in the side-effects between the two agents although more patients taking Golytely (45.5%) deemed its taste unacceptable than those taking Citromag (25.9%). The results of the bowel preparation were assessed by two independent radiologists giving scores on the amount of faecal residue and the quality of mucosal coating. The amount of faecal residue was less in the Golytely group (p < 0.05). The quality of mucosal coating by barium was also better in the Golytely group than the Citromag group (p < 0.05). PMID- 9404203 TI - Comparison of in vivo cardiac function with ex vivo cardiac performance of the rat heart after thoracic irradiation. AB - The aim of the study was to compare in vivo cardiac function with ex vivo cardiac performance after local heart irradiation in the same rat. Left ventricular ejection fraction (LVEF) was measured in vivo by radionuclide ventriculography in Sprague-Dawley rats up to 16 months after a single dose of 20 Gy. Four days after in vivo measurements, cardiac performance was determined ex vivo, using the isolated working rat heart preparation. After irradiation, cardiac performance measured ex vivo deteriorated more rapidly than the in vivo measured LVEF. Within 4 months post-treatment, ex vivo cardiac output and stroke volume started to decrease and declined continuously throughout the observation period of 16 months. The reduction in stroke volume was already significant (p < 0.04) at 4 months post-treatment, whereas the decline in cardiac output was significant (p < 0.05) at 12 months post-treatment. In vivo, no change in LVEF was observed during the first 12 months post-treatment. Thereafter, LVEF decreased rapidly from 65 +/ 2% to 46 +/- 8% (p < 0.01), at 16 months post-treatment. Up to 12 months post irradiation, LVEF was not correlated to ex vivo cardiac output. At 16 months post treatment, when clinical symptoms of heart failure become evident, a positive relation between both parameters was found. The lack of correlation between the in vivo and ex vivo measurements of cardiac function during the first 12 months post-treatment might be explained by the involvement of compensatory mechanisms being operative in vivo to maintain sufficient cardiac output. PMID- 9404204 TI - Reference data for ultrasonic bone measurement: variation with age in 2087 Caucasian women aged 16-93 years. AB - Data from the measurement of broadband ultrasonic attenuation (BUA) and speed of sound (SOS), using the Lunar Achilles ultrasonic densitometer, were collected for Caucasian women from five centres in the United Kingdom (Leeds, London, Nottingham, Lincoln and Sheffield). After correcting for machine variability at each site, the data were combined into a central reference database comprising 2087 women aged 16-93 years. The data are presented in 5-year bands and show a mean fall of 0.36% per year for BUA and 0.08% per year for SOS in the 60 years following the attainment of peak bone mass. This fall in BUA compares with that observed using dual energy X-ray absorptiometry studies of the lumbar spine and femoral neck of 0.32% per year and 0.44% per year, respectively, for the age range 25-65 years. PMID- 9404205 TI - The relationship between breast asymmetry, breast size and the occurrence of breast cancer. AB - Breast cancer is the second most common cancer among women in the world and in developed countries it is the most common. The early identification of women at risk is therefore of great importance and any additional measures which may aid diagnosis, particularly in high risk groups, would be of benefit. Breast volume and breast asymmetry were calculated from mammograms of 250 women with breast cancer and compared with those of 250 age-matched controls. There was evidence that breast cancer patients had more breast asymmetry and larger breasts than age matched healthy women. The former observation is the first evidence that high breast asymmetry may be a risk factor for breast cancer. Breast asymmetry is likely to be a predictor of, rather than the effect of breast cancer. PMID- 9404206 TI - A comparison of the effectiveness of 28 kV (grid) versus 25 kV (no grid) mammographic techniques for breast screening. AB - The purpose of this study was to compare the clinical effectiveness of breast screening using a mammographic technique of 25 kV without a grid, with one of 28 kV with a grid. Effectiveness is judged by cancer detection, interval cancer rates, sensitivity and specificity calculations and tumour characteristics. The doses on standard physics tests given by the three machines when these exposure factors are selected were compared to see whether there is any scientific basis for recommendations on which is the more effective technique. The experiment was undertaken in the prevalence round of a screening programme set up in the UK in 1987. The main comparison is on 25,078 women randomized to one or other technique after March 1989. Comparison can also be made with the preceding 8482 women, who were examined by the 25 kV method, but not randomized and in whom there were a variety of other differences. In the randomized group there was no statistically significant difference in cancer detection. A minor difference in overall numbers without statistical significance was seen in favour of the 28 kV grid technique, but is offset by a greater interval cancer rate in this group. Small cancer detection was equal in the two groups. By contrast, the first 8482 women showed significantly worse screening performance, both in lower overall and small cancer detection rate, and in increased number of interval cancers, for which the explanation is likely to be complex. The dose measurements show that the use of a higher tube potential with the grid mitigates the dose increase that may have been expected. The choice between these two techniques is therefore neither automatically made by greater cancer detection nor made on grounds of dose. There was a minor dose penalty in using the 28 kV technique with grid. PMID- 9404207 TI - The identification of bias in studies of the diagnostic performance of imaging modalities. AB - The demand for evidence-based healthcare is increasing nationally and internationally and it is equally necessary in both diagnostic and therapeutic practice. Evidence may be collected and combined by means of a systematic literature review of published and unpublished data on a well-defined topic. The output of such reviews is then available to guide health policy, influence good practice or direct research. Published guidelines are available on the performance of systematic reviews, especially those of randomized controlled trials. Although there is an extensive literature base of research data in diagnostic imaging there are few such trials, but it is still possible to perform systematic reviews. With the alternative study designs encountered it is important to be aware of the main threats to study validity. In this paper the biases likely to be encountered in studies of diagnostic performance are reviewed, with particular reference to diagnostic imaging tests. The biases are sub-divided into three categories. The first category is patient selection and covers the validity of generalizing results beyond the study population. The other two, concerning study design and execution and the interpretation of results, affect the likely validity of the results of a study. An understanding of these factors is an essential prerequisite for those undertaking or using a systematic literature review in the field of diagnostic imaging. The definitions form the foundations of a defensible review protocol. PMID- 9404208 TI - Effect of automatic kV selection on dose and contrast for a mammographic X-ray system. AB - The effect of automatic tube potential (kV) selection on breast dose and contrast has been assessed using a Philips MammoDiagnost 3000 mammography X-ray set. The performance of the X-ray set using automatic kV selection has been compared with that found using a fixed kV of 28. The AUTOKV mode selected 25 kV for breasts with thickness up to about 50 mm, which increased the contrast by 5-10%, and increased the mean glandular dose (MGD) per film by, on average, 30-40%. For large breasts with a compressed thickness of 70 mm and above, kVs up to 30 were selected so that the average MGD per film was reduced by 19% from 3.62 to 2.94 mGy, with an estimated loss in contrast of about 4-8%. For all breasts the mean MGD per film was 1.85 +/- 0.05 mGy where AUTOKV was used, and 1.74 +/- 0.08 mGy per film when 28 kV was used. The overall image quality of the mammograms was found to be higher when AUTOKV was used. Overall, the AUTOKV facility on this X ray set generally worked well and resulted in slightly higher contrast and slightly better image quality at the price of a small increase in the average dose for this patient group when compared with the usual UK procedure of using a fixed 28 kV. PMID- 9404209 TI - The agreement between colour Doppler systems in measuring internal carotid artery peak systolic velocities. AB - The study was undertaken to determine if the internal carotid artery peak systolic velocities (ICA PSVs) measured by two colour Doppler imaging systems (Acuson 128 and Siemens Quantum) agree sufficiently for the two systems to be interchangeable in evaluating carotid artery disease. One operator obtained blinded measurements of ICA PSV in 63 prospective nonrandomized patients at risk of stroke. The operator examined 20 patients in the first cohort to assess the intraobserver variation, and 43 patients in the second cohort to assess the limits of agreement between the systems. In vitro comparisons of the systems were also undertaken, using both string and flow phantoms. Excluding one outlier, the intraobserver reproducibility coefficient for both machines was 0.48 m s-1. The limits of agreement (within which 95% of differences lie) between systems were 0.47 to 0.45 m s-1. This reduced to -0.39 to 0.33 m s-1 when the one outlier was excluded. This is within the intraobserver reproducibility range. In vitro data show little intersystem variation with phantom velocity. Intratransducer differences increase when the Doppler angle is increased using the string phantom; maximum differences: Acuson 0.30 m s-1 (42%) and Siemens 0.32 m s-1 (32%). These are within the in vivo reproducibility range. Intratransducer difference decreases when the Doppler angle is increased using the flow phantom, maximum differences: Acuson 0.05 m s-1 and Siemens 0.07 m s-1. The results show the systems agree sufficiently to be interchangeable in evaluating carotid artery disease; however, errors in maximum PSVs, caused by operator or system variation, may lead to errors in percent stenosis grading of the carotid arteries. PMID- 9404210 TI - Hawthorne effect: shortening of fluoroscopy times during radiation measurement studies. AB - Screening times were recorded before (n = 92, 13 radiologists) and after (n = 75, 6 radiologists) commencing a protocol with dose-area product (DAP) measurements and filling of structured questionnaires. Fluoroscopy times were significantly (p = 0.0001) longer before starting these measurements (median 4.3, mean 5.2 min) than during them (median 3.2, mean 3.6 min), which indicates a Hawthorne effect. Fluoroscopy times did not increase during the DAP measurement period up to 21 barium enemas and a study period of up to 45 days per radiologist. Previous fluoroscopic radiation measurements based on action during an analysis period may be biased towards too short fluoroscopy times and too low doses. Radiation measurement, even if not scientifically indicated, seems a practical way of reducing doses. PMID- 9404211 TI - Three phase 99Tcm (V)DMSA scintigraphy in Paget's disease: an indicator of pamidronate effect. AB - The use of three phase 99Tcm (V)DMSA scintigraphy is reported in a patient with Paget's disease of bone before and after intravenous pamidronate therapy. It was a useful modality for estimating the activity of Pagetoid lesions and the therapeutic effect of pamidronate, from a different aspect to bone scintigraphy. Three phase 99Tcm (V)DMSA scintigraphy evaluates both the blood flow and the metabolic activity of Pagetoid bone. PMID- 9404212 TI - Visualization of central stellate fibrosis in hyaline vascular type Castleman's disease. AB - Hyaline vascular type Castleman's disease (HVCD) is a benign cause of lymph node hypertrophy which radiologically resembles tumours such as paraganglioma and lymphoma. However, a distinguishing pathological characteristic of HVCD is the presence of central stellate fibrosis. Since conventional CT and MRI scans failed to differentiate areas of fibrosis, three-phase dynamic CT was performed on a patient who was thought to have HVCD. The images showed central fibrosis and consequently the correct pre-operative diagnosis of HVCD was made. PMID- 9404213 TI - Parapharyngeal space lipoma causing sleep apnoea. AB - Lipoma of the parapharyngeal space is very rare, only three cases having been reported in the literature. A parapharyngeal space lipoma causing obstructive sleep apnoea has not been reported before. A 60-year-old man presented at the ear, nose and throat (ENT) clinic with a history of loud snoring associated with sleep apnoea secondary to a right parapharyngeal space lipoma. The causes of sleep apnoea and the radiological features of a parapharyngeal space lipoma are discussed. PMID- 9404214 TI - Fistula formation from neovascularity developing in malignant fibrous histiocytoma of the heart. AB - Neovascularity in cardiac tumours is uncommon and fistula formation from such neovascularity is extremely rare. Fistula formation from neovascularity developing in a malignant fibrous histiocytoma of the heart is described in a 48 year-old man. The the case illustrates that cardiac tumour is not only a cause of cardiac output obstruction but also of coronary artery fistula. PMID- 9404215 TI - Uterine lipoleiomyoma: MRI, CT and ultrasonographic findings. AB - MRI, CT and ultrasonographic findings in lipoleiomyoma of the uterus are reported. A 73-year-old woman presented with a palpable mass in the lower abdomen. T2 weighted MR images showed an 8 x 5 cm exophytic well defined hyperintense mass of the uterine body. Fat content within the tumour was confirmed because of evident chemical shift artefact. Microscopically, the tumour proved to be lipoleiomyoma consisting of smooth muscle cells and mature adipose tissue. Five cases have been reported in the imaging literature, and the presence of fat within a uterine mass on CT or MRI is diagnostic of lipoleiomyoma. PMID- 9404216 TI - CT scanning of the paranasal sinuses--the effect of reducing mAs. AB - High quality CT scans are required prior to fibreoptic endoscopic sinus surgery (FESS) surgery and in many institutions such scans are performed using a high mAs technique. Consequently, the investigation imparts a radiation dose to the patient and in particular to the eye. Such a radiation dose is a possible source of morbidity. We believe that the mAs, and consequently the radiation dose, can be considerably reduced without affecting scan quality. The present study compares the quality of sinus CT scans performed at two mAs values, 40 and 60. Scan quality was assessed in terms of the ability to visualize clearly important anatomical structures and in terms of overall perceived quality. We show that mAs values as low as 40 can be used without adversely affecting the diagnostic quality of the examination. PMID- 9404217 TI - A leak and a dowel movement. PMID- 9404218 TI - Haemoptysis: a rare cause. PMID- 9404219 TI - Glycoprotein changes in tumours: a renaissance in clinical applications. AB - 1. Oligosaccharides linked to protein (glycoprotein) or lipid (glycolipid) are the major components at the outer surface of mammalian cells. Studies using antibodies and lectins have shown in the past that the oligosaccharides they recognize exhibit tumour-associated changes, i.e. they are carbohydrate tumour associated antigens. 2. The oligosaccharides have been further characterized in recent years by structural analysis using high-resolution chromatographic techniques, MS and NMR. NMR gives an oligosaccharide finger-print that is characteristic of monosaccharide type and linkage and which can be correlated with magnetic resonance spectroscopic data on fine-needle tissue aspirates. 3. Also of relevance is the new understanding of the molecular biology of MUC genes, which code for mucin protein backbones, and of the glycosyltransferase genes, which determine oligosaccharide structure and immunological recognition. 4. For these reasons, we believe that tumour-associated oligosaccharide changes should be revisited in the context of what we now know about structure and expression. This review synopsizes the past data using the detection of carbohydrate tumour associated antigens by binding of lectins and antibodies, and puts it into the context of NMR fingerprints or signatures. PMID- 9404220 TI - Activation of two inward chloride transport systems in rat femoral arterial smooth muscle in deoxycorticosterone acetate/salt hypertension. AB - 1. Intracellular [Cl-] ([Cl-]i) was measured with ion-selective microelectrodes in rat femoral arterial smooth muscle in normotensive controls and after the induction of deoxycorticosterone acetate/salt hypertension. 2. Linear regression of [Cl-]i and time after the induction of hypertension showed good correlation (r = 0.96) for 5-6 weeks, as [Cl-]i increased from 30 +/- 1 mmol/l (mean +/- SD, n = 16), to 49 +/- 2 mmol/l (n = 9, P < 0.0001). 3. Arterial systolic blood pressure also increased linearly (r = 0.97) for 5-6 weeks as hypertension developed from 122 +/- 1 mmHg (n = 20) to 187 +/- 7 mmHg (n = 14): there was consequently a linear relationship between [Cl-]i and arterial systolic blood pressure (r = 0.96). 4. The increase in [Cl-]i was partly because Na(+)-K(+)-Cl- co-transport activity, estimated from the fall in [Cl-]i caused by bumetanide, was greater in hypertension (18 mmol/l) than in normotension (10 mmol/l). This finding, and the depolarization of the membrane potential in hypertension (-56 +/- 3 mV compared with -64 +/- 4 mV in normotension; P < 0.0001), confirms previous studies. 5. The increase in [Cl-]i was also partly due to greater activity of an Na(+)- and HCO3( )-independent, acetazolamide-sensitive inward Cl- transport system; thus acetazolamide reduced [Cl-]i by 7 mmol/l in normotension and by 16 mmol/l in hypertension. 6. In Cl(-)-free media, the membrane potential in normotension (-59 +/- 5 mV) was not significantly different from that in hypertension (-60 +/- 4 mV). 7. The role of [Cl-]i in the depolarization of the membrane potential in hypertension is discussed. PMID- 9404221 TI - Age, gender and fractal scaling in heart rate variability. AB - 1. The fractal scaling of heart rate variability, gauged by the correlation dimension (CD), is hypothesized to be characterized by a time structure (chronome), which in health shows differences as a function of gender and age. 2. From 24 h Holter records of 44 clinically healthy male subjects in four age groups (5-10, 20-25, 40-45 and 60-65 years; n = 11 in each group), 500 s sections at 4 h intervals for 24 h were analysed for smoothed R-R intervals sampled at 4 Hz. Using an algorithm modified from Grassberger and Procaccia (Physica D 1983; 9: 189-208), the correlation integral was estimated for embedding dimensions from 1 to 20 with a 1.0 s time lag for each section. Nightly (02.00 hours-06.00 hours) ECG records were similarly analysed in 72 additional clinically healthy subjects of both genders, 5-70 years of age. The single cosinor assessed the circadian characteristics; one- and two-way analyses of variance and linear regression were used to examine changes as a function of gender and age. 3. The 24 h average of CD is largest in the 20-25-year-old men and decreases with age there-after (P < 0.05). These changes apply in particular to the nightly CD values, which are higher in female than in male subjects (P < 0.001). Increasing age is associated with a decrease in the amplitude and an advance in the phase of the circadian rhythm in CD (P < 0.05). 4. A chaotic end-point from fractal scaling, yielding a non-linear index, such as the correlation integral, undergoes a circadian rhythm and changes with gender and age. This assessment in the chronome represents an added diagnostic tool in cardiology, and provides new end-points for the study of coherence among internal variables of autonomic mechanisms and of influences by external environmental variables upon them. PMID- 9404222 TI - Age- and gender-related changes in endothelin and catecholamine release, and in autonomic balance in response to head-up tilt. AB - 1. There is an increase in circulating levels of vasoconstrictive hormones and an alteration in baroreceptor responsiveness with aging. The role of changes in endothelium-dependent and -independent vasoconstrictive hormones in relation to age and gender, with simultaneous assessment of autonomic balance in response to head-up tilt, has been incompletely studied. 2. Sixteen young [25 +/- 3 years (mean +/- SEM)] and 16 older normal volunteers (68 +/- 7 years) underwent a 30 min head-up tilt test at 60 degrees. Haemodynamics were measured every 5 min and blood samples for neurohormone measurement were drawn at baseline, 5, 10, 15 and 30 min into the test. Heart rate variability was analysed in 5 min segments at the baseline, and during the test. The younger subjects exhibited a greater increase in heart rate and diastolic blood pressure, despite lower absolute levels of noradrenaline (norepinephrine) and endothelin-1. Analysis of heart rate variability yielded a decrease in both high- and low-frequency bands in the aged; power at low-frequency decreased only in the young subjects. The age-related differences in blood pressure and noradrenaline levels were markedly attenuated in the female subjects. In addition, endothelin-1 levels and power spectral measurements at low frequency were the lowest in younger females throughout the tilt. 3. Despite attenuated cardiovascular response to tilt, both systemic adrenergic 'drive' and endothelin-1 levels increase in parallel with aging. Thus, endothelium-dependent and -independent vasoconstrictive hormone levels increase with age in the resting state and in response to neurohumoral stimulation in humans. PMID- 9404223 TI - Reproducibility of ultrasonographic measurements of different carotid and femoral artery segments in healthy subjects and in patients with increased intima-media thickness. AB - 1. The reproducibility of measurements of the arterial wall thickness in both the carotid and femoral artery was investigated by means of high-resolution B-mode ultrasonography. For this purpose, subjects with normal and increased intima media thickness were selected. Images were stored on an optical disk and were analysed with a semi-automatic software program by two readers. Individuals were scanned twice by two independent observers. 2. Measurements were performed of the far and near wall of the common carotid artery and bulbous in 30 healthy subjects and 19 patients known to have an increased intima-media thickness. Far-wall measurements were made of the internal carotid artery on both sides and common femoral artery on the right side only. 3. In healthy subjects the mean within observer coefficient of variation was 1.8% and 3.0% for the far wall in the common carotid artery on the right side and left side, respectively. For the near wall the mean coefficient of variation of the common carotid artery was 2.8% on the right and 3.4% on the left side. The mean coefficient of variation was less than 4% for both far and near wall in the bulbous and far wall in the internal carotid artery. Even in patients with increased intima-media thickness the mean coefficient of variation of each segment was less than 4.5%. In the control subjects the between-observer coefficient of variation of the common carotid artery was 2.8% and 5.1% for the far wall on the right and left side, respectively, and 3.4% and 4.2% for the near wall on the right and left side. In healthy subjects a mean difference of 0.002 mm within observers was found in the right far-wall common carotid artery, with limits of agreement of -0.048 to 0.052 mm. The coefficient of repeatability was 0.050 mm. For patients with increased intima-media thickness the mean difference in this segment was -0.006 mm (-0.094 to 0.082) with a coefficient of repeatability of 0.088 mm. For the near wall in the common carotid artery and far and near wall in the bulbous and internal carotid artery the mean differences were larger, but were all below 0.1 mm. The differences and limits of agreements increased between observers. In patients the between-observer mean difference of the far wall of the common carotid artery was -0.055 mm (-0.255 to 0.145). For the common femoral artery of normal control subjects the within- and between-observer mean differences were 0.005 mm (-0.119 to 0.129) and 0.015 mm (-0.081 to 0.111), respectively. 4. In conclusion, the reproducibility of intima-media thickness measurements in the common carotid artery is reliable, even in patients with increased artery wall thickness. Also in other segments prone to atherosclerosis, such as the bulbous, internal carotid artery and common femoral artery, a good reproducibility was found. To obtain good reproducibility it is highly recommended to use the same ultrasonographer to scan patients in follow-up studies. PMID- 9404224 TI - Evaluation of QT interval length, QT dispersion and myocardial m iodobenzylguanidine uptake in insulin-dependent diabetic patients with and without autonomic neuropathy. AB - 1. An association has been reported between QT interval abnormalities and cardiovascular autonomic neuropathy in diabetic patients. The QT interval abnormalities reflect local inhomogeneities of ventricular recovery time and may be related to an imbalance in cardiac sympathetic innervation. Sympathetic innervation of the heart can be visualized and quantified by single-photon emission-computed tomography with m-[123I]iodobenzylguanidine. In this study we evaluated cardiac sympathetic integrity by m-[123I]iodobenzylguanidine imaging and the relationship between both QT interval prolongation and QT dispersion from standard 12-lead ECG variables and m-[123I]iodobenzylguanidine uptake in insulin dependent diabetic patients. 2. Three patient groups were studied, comprising six healthy control subjects, nine diabetic patients without cardiovascular autonomic neuropathy (CAN-) and 12 diabetic patients with cardiovascular neuropathy (CAN+). Resting 12-lead ECG was recorded for measurement of maximal QT interval and QT dispersion. The QT interval was heart rate corrected using Bazett's formula (QTc) and the Karjalainen approach (QTk). Quantitative measurement (in counts/min per g) and visual defect pattern of m-[123I]iodobenzylguanidine uptake were performed using m-[123I]iodobenzylguanidine single-photo emission-computed tomography. 3. Global myocardial m-[123I]iodobenzylguanidine uptake was significantly reduced in both diabetic patient groups compared with control subjects. The visual defect score of m-[123I]iodobenzylguanidine uptake was significantly higher in CAN+ diabetic patients than in control subjects and in CAN- patients. This score was not significantly different between control subjects and CAN- patients. QTc interval and QT dispersion were significantly increased in CAN+ diabetic patients as compared with control subjects (QTc: 432 +/- 15 ms versus 404 +/- 19 ms, P < 0.05; QT dispersion: 42 +/- 10 versus 28 +/- 8 ms, P < 0.05). QT dispersion was also significantly longer in CAN- diabetic patients than in control subjects (41 +/- 9 ms versus 28 +/- 8 ms, P < 0.05). QTc interval was significantly related to global myocardial m-[123I]iodobenzylguanidine uptake and defect score in diabetic patients (r = -0.648, P < 0.01, and r = 0.527, P < 0.05, respectively). There was no correlation between QT dispersion and both m-[123I]iodobenzylguanidine uptake measures. 4. In conclusion, these findings suggest that m [123I]iodobenzylguanidine imaging is a valuable tool for the detection of early alterations in myocardial sympathetic innervation in long-term diabetic patients without cardiovascular autonomic neuropathy. Insulin-dependent diabetic patients with cardiovascular autonomic neuropathy have a delayed cardiac repolarization and increased variability of ventricular refractoriness. The cardiac sympathetic nervous system seems to be one of the determinants of QT interval lengthening, but does not appear to be involved in dispersion of ventricular recovery time. It is assumed that QT dispersion is based on more complex electrophysiological mechanisms which remain to be elucidated. PMID- 9404225 TI - Mutations at the lipoprotein lipase gene locus in subjects with diabetes mellitus, obesity and lipaemia. AB - 1. The common association of obesity, diabetes mellitus and hyperlipidaemia may have a primary aetiological basis. Insulin resistance has been postulated as a possible cause, but defects in the plasma transport of triacylglycerol or fatty acids could also be primary determinants. 2. We have therefore studied 18 patients with diabetes mellitus, obesity and severe hypertriglyceridaemia for defects of a key protein involved in the clearance of plasma triacylglycerols, lipoprotein lipase. 3. DNA was prepared from leucocytes of 18 patients with the above syndrome, and exons encoding lipoprotein lipase were amplified by PCR. The products were sequenced using the dideoxy chain-termination method. 4. Eight of the subjects were found to possess genetic variants at the lipoprotein lipase gene locus. These were: (a) G579-->A, V108V; (b) G818-->A, G188E; (c) C829-->T, R192; (d) A1127-->G, N291S; (e) C1308-->G, F351L; (f) C1338-->A, T361T; and (g) C1595-->G, S447. Three of these, (c), (e) and (f), have not hitherto been described. Variant (f), appears to be a population polymorphism whose allele frequency in normolipidaemic diabetics was found to be 0.12 (162 chromosomes studied). The others are all rare at frequencies of < 0.01 and may contribute to the phenotype by impairing clearance of plasma triacylglycerols. 5. We conclude that genetic variants at the lipoprotein lipase locus occur commonly in subjects with this syndrome (four out of 18 subjects with probably functional mutants) and may affect the individual's response to obesity and diabetes mellitus for the development of lipaemia. PMID- 9404226 TI - Elevated post-prandial gastric inhibitory polypeptide concentrations in hypertriglyceridaemic subjects. AB - 1. We investigated whether abnormalities of gastric inhibitory polypeptide (GIP) and glucagon-like peptide-1 (7-36 amide) (GLP-1) contribute to the hypertriglyceridaemia and hyperinsulinaemia in hypertriglyceridaemic subjects. Serum triglycerides and plasma glucose GIP, GLP-1 and immunoreactive insulin (IRI) concentrations were measured before and after a mixed meal in 15 hypertriglyceridaemic patients and in eight healthy normotriglyceridaemic control subjects. 2. Integrated post-prandial GIP concentrations were greater than in controls (P < 0.05) and correlated positively with both fasting and integrated post-prandial triglyceride concentrations (P < 0.05 for both). Fasting and integrated post-prandial IRI levels were higher in hypertriglyceridaemic subjects than in controls (P < 0.02 and P < 0.05 respectively) and correlated positively with fasting triglycerides (P < 0.02 and P < 0.001 respectively) and integrated post-prandial triglycerides (P < 0.005 and P < 0.05 respectively). There was no correlation between GIP concentrations and either fasting or post-prandial IRI levels. Fasting and post-prandial concentrations of GLP-1 were similar in patients and controls. 3. Hypertriglyceridaemic subjects have post-prandial hyperGIPaemia in addition to the well-documented hyperinsulinaemia. We found no association between GIP and insulin. There is, however, clear evidence for an association between post-prandial GIP concentrations and triglyceride levels. We suggest that this association may depend on changes in lipoprotein lipase activity and that there may be a feedback loop between GIP and triglyceride lipolysis. PMID- 9404227 TI - Brown adipose tissue and its uncoupling protein in chronically hypoxic rats. AB - 1. Hypoxia is known to decrease thermogenesis. We set out to determine whether this is accompanied by alterations in the brown adipose tissue, which is a major source of non-shivering thermogenesis. 2. Measurements were performed on 25- and 64-day-old rats, after 4 days of hypoxia (10% inspired O2), and on approximately 3.5-month-old rats in hypobaric hypoxia since birth, at an ambient temperature of 25 degrees C. 3. All hypoxic rats had higher haematocrit and lower body mass than corresponding controls. 4. In the 25-day-old rats, hypoxia had minimal and non significant effects on brown adipose tissue mass, proteins and DNA concentration. The content of the mitochondrial uncoupling protein thermogenin, evaluated by immunoblot after electrophoretic separation, relative to the cytoskeleton actin (UCP/Act), was not significantly altered. 5. In 25-day-old rats exposed for 4 days to cold (ambient temperature = 7-9 degrees C), brown adipose tissue was hyperplastic, with increased UCP/Act; hypoxia did not appreciably alter the response to cold. 6. In the 2-month-old rats, after 4 days of hypoxia UCP/Act was reduced to about 40% of control. 7. In the 3.5-month-old rats maintained in hypoxia since birth, brown adipose tissue mass was reduced in proportion to body mass, with little effect on total proteins and DNA; UCP/Act was decreased to about 50% of control. 8. We conclude that chronic hypoxia had a minimal effect on brown adipose tissue total proteins and DNA content. However, the uncoupling protein content can be greatly reduced, depending upon age and duration of hypoxia. PMID- 9404228 TI - Selective migration of alpha-smooth muscle actin-positive myofibroblasts toward fibronectin in the Boyden's blindwell chamber. AB - 1. In order to address the hypothesis that migrating fibroblasts have a different phenotype, human fetal lung fibroblasts (HFL-1) cells were evaluated in the Boyden blindwell chamber migration assay followed by immunoelectron microscopy. 2. HFL-1 cells were placed on nucleopore filters and incubated for 2 h using purified human plasma fibronectin (pFn) as a chemoattractant. Filters were then processed for immunoelectron microscopy using antibodies for alpha-smooth muscle (alpha-SM) actin as a marker for myofibroblasts, cellular fibronectin (cFn) and VLA-5. 3. Cells which had migrated to the bottom side of the filter were more likely to express alpha-SM actin, 29.1 +/- 3.4% of cells, compared with cells which did not migrate through the filter, 12.4 +/- 1.3% (P < 0.05). The total proportion of alpha-SM-actin-positive cells located on both sides of the filter showed no difference between those which had migrated toward pFn and controls (17.7 +/- 1.0% compared with 20.2 +/- 2.5%). 4. cFn-positive cells showed minimal differences compared with control cells, while perinuclear and endoplasmic reticulum staining of VLA-5 was observed only in the cells treated with pFn. 5. The results show that HFL-1 cells are heterogenous for alpha-SM actin expression. Short-term incubation with pFn did not change the proportion of alpha-SM-actin positive HFL-1 cells. Cells which migrate, however, are enriched for alpha-SM actin expression. pFn-induced fibroblast chemotaxis can selectively recruit myofibroblasts with increased alpha-SM actin expression, a feature which may contribute to the altered population of cells at sites of fibrosis. PMID- 9404229 TI - Mast cell activation in arthritis: detection of alpha- and beta-tryptase, histamine and eosinophil cationic protein in synovial fluid. AB - 1. Although mast cell hyperplasia is a feature of rheumatoid arthritis and osteoarthritis, the extent and nature of mast cell activation in joint disease have not been clearly established. 2. We have investigated the levels of mast cell tryptase and histamine and also of eosinophil cationic protein in synovial fluid collected from 31 patients with rheumatoid arthritis, 14 with seronegative spondyloarthritis and nine with osteoarthritis. Two RIAs for tryptase were employed: one with monoclonal antibody AA5, which was found to bind equally well to both alpha and beta isoforms on Western blots of the recombinant enzyme, and the other with antibody G5, which recognizes predominantly beta-tryptase. 3. alpha-Tryptase, which is likely to be released constitutively from mast cells, appeared to be the major form in synovial fluid, as the assay with antibody AA5 detected appreciably more tryptase than that with antibody G5. beta-Tryptase, which is released on anaphylactic activation of mast cells, was detected in 14 out of 45 synovial fluid samples studied, with concentrations of up to 12 micrograms/l measured by the G5 assay. The apparent levels of beta-tryptase, but not of alpha-tryptase, were closely correlated with those of histamine in the synovial fluid. Patients with osteoarthritis appeared to have a greater proportion of beta-tryptase in the synovial fluid than those with rheumatoid arthritis, as well as higher concentrations of histamine. Eosinophil cationic protein was present at high levels in the synovial fluid, although eosinophil numbers were low, and its concentrations were not correlated with the concentrations of the mast cell products. 4. These data suggest that anaphylactic degranulation of mast cells may have occurred to a greater extent in osteoarthritis than in rheumatoid arthritis, despite the relative lack of synovial inflammation in osteoarthritis. Although the eosinophil cationic protein detected may not reflect eosinophilic inflammation in the joint, the presence in synovial fluid of tryptase of both major forms, and of histamine, appears to indicate that mast cell products are secreted constitutively, as well as by processes of anaphylactic degranulation in rheumatoid arthritis, seronegative spondyloarthritis and osteoarthritis. PMID- 9404230 TI - A single dose of intravenously injected poloxamine-coated long-circulating particles triggers macrophage clearance of subsequent doses in rats. AB - 1. Adsorption of the block copolymer non-ionic surfactant poloxamine-908 on to the surface of polystyrene nanospheres (60 nm in diameter) produced 'phagocyte resistant' particles (otherwise known as long-circulating particles). This was reflected by a profound reduction in uptake of such engineered nanospheres by macrophages of the reticuloendothelial system and extended blood circulation time, after intravenous administration to rats. 2. A single intravenous administration of poloxamine-908-coated particles dramatically affected the circulation half-life and body distribution of a second subsequent dose. The degree of alteration depended on the interval between the two doses. At 3 days after a single intravenous injection of poloxamine-coated particles, Kupffer cells and spleen macrophages could clear a second dose of long-circulating beads from the blood. When tested at day 14, the second dose of intravenously injected poloxamine-coated particles avoided rapid uptake by liver and spleen macrophages and remained in the blood. 3. The coating polymer (poloxamine-908) apparently triggered bead clearance by resident Kupffer cells and certain sub-populations of spleen macrophages, since a single intravenous dose of an endotoxin-free solution of poloxamine 3 days before the administration of long-circulating particles induced similar effects. When the interval between the two injections was 2 weeks, poloxamine-coated particles again exhibited long circulation half-life. This cycle could be repeated after intravenous administration of a second poloxamine dose 2 weeks after the first poloxamine injection. 4. The mechanism of particle recognition by resident tissue macrophages was found to be independent of opsonization processes. 5. These studies could have important implications in biomedical application, design and engineering of poloxamine-based long circulating drug carriers for repeated intravenous administration. PMID- 9404231 TI - Prognosis of chronic hepatitis C patients correlates with circulating monocyte/macrophage function. PMID- 9404232 TI - New clinical strategies for symptom management and quality of life enhancement in cancer patients. PMID- 9404233 TI - Cancer-specific quality of life questionnaires: the state of the art in Europe. AB - Quality of life (QL) assessment is now regarded as desirable, if not mandatory, by agencies supporting cancer clinical trials around the world, yet doubts persist about the relevance of QL data to clinical practice. A plethora of QL measures is available but the quality of published work remains suboptimal. The appropriate choice of instrument is essential if outcome measures are to be valid and clinically meaningful. This paper reviews the considerations which should determine the choice of QL questionnaire and, taking the specific example of the EORTC approach, aims to provide users with an update on the current state of the art in the development of cancer-specific QL questionnaires. PMID- 9404234 TI - New approaches to pain control in patients with cancer. AB - Pain affects most patients with malignant disease, and the prevalence of severe pain increases in the advanced stages of the condition. One in 5 patients with cancer has uncontrolled pain, even after 10 years of the use of the World Health Organization programme for cancer pain control and its 'three-step ladder' for the rational use of analgesics including morphine. Morphine has long been the 'gold standard' for the treatment of severe cancer pain. However, its side effects, particularly sedation, cognitive impairment and myoclonus at high doses, have provoked the use of 'opioid rotation' to alternatives such as methadone and hydromorphone. The new 72-h transdermal patch for fentanyl also offers advantages of reduced side-effects and increased convenience over oral morphine. Intravenous strontium-89 and bisphosphonate therapy are effective for both short- and long term control of metastatic bone pain. The spinal N-methyl-D-aspartate (NMDA) receptor is important in modulating the plasticity of the central nervous system and in aggravating chronic pain through the phenomenon of 'wind-up'. The NMDA antagonist ketamine, an anaesthetic, can be used at low doses for the management of refractory and neuropathic pains. Among adjuvant drugs, ketorolac has emerged as a potent non-steroidal anti-inflammatory drug. Palliative care is gaining acceptance as a new discipline in healthcare. Its strategic role is being reviewed as an adjunct to cancer therapy at all stages and its use is no longer confined to the terminal phase of disease after curative treatment has failed. Pain control and other aspects of symptom control are, therefore, viewed as an integral part of cancer management. PMID- 9404235 TI - Communication skills and psychological training in oncology. AB - Preserving the best possible quality of life for cancer patients and their families has become a major goal in cancer care. However, the cumulative effect of stressors related to cancer care, many of which involve communicating with patients and relatives, may lead to the development of burnout in staff. Many health care professionals lack the psychosocial knowledge and communications skills needed to identify patients' problems because general professional training focuses on technical care. Teaching strategies known as psychological training programs (PTP) are therefore being developed to help improve health care professionals' sensitivity to communication problems with patients and relatives. Cognitive (e.g. theoretical information), experiential (e.g. case-history discussions), behavioural (e.g. role-playing exercise) and supportive (e.g. stressor identification) training techniques are used to teach the essential skills of good communication, i.e. listening, empathy, response to cues and appropriate use of reassurance. PTP range from one-day courses and residential workshops to full-time 1- or 2-year curricula. However, one of the main obstacles to implementing PTP is scepticism among health care professionals about its usefulness. Research on training effectiveness should therefore be developed to assess the impact of communication skills on quality of care and patients' quality of life. PMID- 9404236 TI - Guidelines for clinical trials in Helicobacter pylori infection. Working Party of the European Helicobacter pylori Study Group. PMID- 9404238 TI - Statistical annex: statistical aspects of clinical trials in Helicobacter pylori infection. Working Party of the European Helicobacter pylori Study Group. PMID- 9404237 TI - Technical annex: tests used to assess Helicobacter pylori infection. Working Party of the European Helicobacter pylori Study Group. PMID- 9404239 TI - Transferring patients for primary angioplasty. PMID- 9404240 TI - Willem Einthoven (1860-1927). PMID- 9404241 TI - Transferring patients for primary infarct angioplasty. PMID- 9404242 TI - When should patients with acute myocardial infarction be transferred for primary angioplasty? PMID- 9404243 TI - Treating coronaries, at home or away? PMID- 9404244 TI - Lessons from myocardial contrast echocardiography studies during primary angioplasty. PMID- 9404245 TI - Transferring patients for primary angioplasty: a retrospective analysis of 104 selected high risk patients with acute myocardial infarction. AB - OBJECTIVE: To investigate the feasibility of primary coronary angioplasty as a treatment option in patients with acute myocardial infarction after initial diagnosis in a local community hospital. SETTING: Referral centre for interventional treatment of coronary artery disease. METHODS: During a five year period, 520 candidates for primary coronary angioplasty were treated in our institution, 104 after transfer from a community hospital. The transferred patients and the non-transferred patients (n = 416) were compared with regard to baseline clinical characteristics, time interval from symptom onset to treatment, and clinical outcome at six months. RESULTS: In this setting, the influence of transportation on total ischaemic time was limited, and there was no difference in clinical outcome between the transferred and the non-transferred patients. Clinical outcome was mainly dependent on the indication for transfer. CONCLUSIONS: Safe and expedient transportation may facilitate the more widespread use of primary angioplasty in patients with acute myocardial infarction. A large randomised multicentre trial is needed to compare the relative merits of intravenous thrombolytic treatment in a local hospital with primary angioplasty after transfer in selected high risk patients with acute myocardial infarction. PMID- 9404247 TI - An unusual palliative shunt for cyanotic congenital heart disease. PMID- 9404246 TI - Restenosis after elective coronary balloon angioplasty in patients with end stage renal disease: a case-control study using quantitative coronary angiography. AB - OBJECTIVE: To assess the rate of angiographic restenosis in patients with end stage renal disease after elective coronary angioplasty. DESIGN: A retrospective case-control study of 20 patients with end stage renal disease and 20 sex and age matched controls without renal disease, who had undergone primarily successful coronary angioplasty. Control coronary angiography was performed regardless of worsening or renewed incidence of anginal symptoms. MAIN OUTCOME MEASURES: Group comparison of coronary morphology, as evaluated by quantitative coronary angiography, and of cardiovascular risk factors. RESULTS: The rate of angiographic restenosis was 60% in patients with renal disease and 35% in controls. In patients with end stage renal disease the following differences (mean (SD) were found versus controls: raised plasma fibrinogen (483 (101) v 326 (62) mg/dl, p < 0.001); raised plasma triglyceride (269 (163) v 207 (176) mg/dl, p < 0.01); smaller diameter of the coronary reference segment (2.59 (0.87) v 2.90 (0.55) mm, p < 0.10); smaller minimum luminal diameter of the dilated stenosis (0.77 (0.46) v 0.97 (0.27) mm, p < 0.05). Discriminant analysis showed that minimum luminal diameter before angioplasty (r = -0.79) and fibrinogen (r = +0.34) had the highest statistical association with restenosis. CONCLUSIONS: The high rate of angiographic restenosis in patients with end stage renal disease seems to be related to the size of the vessel dilated and to an increased prothrombotic risk, as indicated by higher fibrinogen concentrations. PMID- 9404248 TI - Coronary artery bypass graft surgery in dialysis patients. AB - OBJECTIVE: To examine the short term results and long term survival of patients on long term dialysis undergoing coronary artery bypass graft surgery. METHODS: A retrospective analysis of 19 patients on established dialysis who underwent coronary revascularisation between 1983 and 1995; 14 patients (73%) had class IV angina and five (25%) had unstable angina requiring heparin and nitrate infusions before surgery. RESULTS: The 30 day mortality was 5%. Follow up was completed in the remaining 18 patients. The mean follow up time was 34 months (range eight to 61). During the follow up period four patients died of cardiac causes. The actuarial survival at one, two, and three years was 87%, 78%, and 59%, respectively. The overall functional status was significantly improved compared to preoperative levels, with a mean Karnofsky score of 76% (p < 0.01) at three years. CONCLUSIONS: Coronary artery bypass graft surgery can be performed with increased but acceptable morbidity and mortality in chronic dialysis patients. It results in considerable improvement in symptoms and functional status. However, long term survival is limited and this requires further investigation. PMID- 9404249 TI - Dipyridamole thallium-201 scintigraphy for early risk stratification of patients after uncomplicated myocardial infarction. AB - OBJECTIVE: To determine the safety and prognostic value of dipyridamole thallium 201 scintigraphy performed in patients within three to five days of acute myocardial infarction, including those receiving thrombolytic treatment. DESIGN: A prospective study of dipyridamole thallium-201 scintigraphy in patients early after acute myocardial infarction. SETTING: University hospital. PATIENTS: 200 patients who were clinically uncomplicated at day 3 after infarction, 92 (46%) of whom had received thrombolysis. MAIN OUTCOME MEASURES: Incidence of cardiac death, non-fatal reinfarction, readmission to hospital for unstable angina, or non-elective revascularisation procedure within six months' follow up. RESULTS: No patient had a serious complication from the dipyridamole study. At six month follow up, 55 patients (28%) had suffered a defined cardiac event. Patients who received thrombolysis had the same extent of thallium-201 redistribution and the same occurrence of subsequent cardiac events as those not receiving thrombolysis. Patients who subsequently had an event had more myocardial segments showing thallium-201 redistribution than event free patients: 2.7 (SD 1.9) v 1.2 (1.4), respectively (p < 0.001). Among all clinical and scintigraphic variables, multivariate analysis identified the extent of thallium-201 redistribution as the only independent predictor of outcome (p < 0.001). Among 63 patients (32%) of the study cohort who showed more than two myocardial segments with thallium-201 redistribution, the adjusted risk ratio for a cardiac event was 7.5 (95% confidence interval 2.9 to 19.1) compared with patients without any redistribution. CONCLUSIONS: Dipyridamole thallium-201 scintigraphy can be performed safely within a few days of the event in patients with uncomplicated myocardial infarction, including those who received thrombolysis, and can identify a subgroup of patients at high risk of future ischaemic events. PMID- 9404250 TI - Combination treatment with trimetazidine and diltiazem in stable angina pectoris. AB - OBJECTIVE: To assess antianginal efficacy and possible adverse haemodynamic effects of combination treatment with trimetazidine and diltiazem in patients with stable angina. DESIGN: Double blind, randomised, placebo controlled trial of four weeks duration. SETTING: Outpatient department of two Indian hospitals. SUBJECTS: 64 male patients with stable angina, uncontrolled on diltiazem alone. INTERVENTIONS: Diltiazem 180 mg and trimetazidine 60 mg, or diltiazem 180 mg and placebo daily. MAIN OUTCOME MEASURE: Change in exercise time to 1 mm ST segment depression. RESULTS: 33 patients (55%) had no exercise induced angina at 3 mm ST segment depression at inclusion in the study (silent ischaemia). Intention to treat analysis showed that of 32 patients in each treatment group, the number (%) of patients responding to trimetazidine compared to placebo was: for anginal attacks, 28 (87.5) v 15 (46.9), p < 0.001; for exercise time to 1 mm ST segment depression, 21 (65.6) v 9 (28.1), p < 0.003; for exercise time to angina, 12 (37.5) v 5 (15.6), p < 0.05; and for maximum work at peak exercise, 17 (53.1) v 8 (25), p < 0.02. Compared to placebo, there was net improvement with trimetazidine in mean anginal attacks of 4.8/ week (95% confidence interval (CI) 7.5 to 2.1; p < 0.002); in mean exercise times at 1 mm ST segment depression of 94.2 seconds (95% CI 182.8 to 5.6; p < 0.05), and at onset of angina of 113.1 seconds (95% CI 181.6 to 44.6; p < 0.02); and in mean maximum work at peak exercise of 1.4 metabolic equivalents (95% CI 2.4 to 0.3; p < 0.05). CONCLUSIONS: Patients with stable angina uncontrolled with diltiazem had a clinically important improvement after combination treatment with trimetazidine, without adverse haemodynamic events or increased side effects. PMID- 9404251 TI - Effects of atrioventricular asynchrony on platelet activation: implication of thromboembolism in paced patients. AB - OBJECTIVE: To investigate the platelet activation in different modes of pacing in patients implanted with dual chamber rate adaptive pacemaker (DDDR) for bradyarrhythmias, and to explore the possible underlying mechanism of the higher thromboembolic incidence in single chamber ventricular rate adaptive (VVIR) pacing. DESIGN: Platelet activation was determined in chronically paced patients during three different pacing modes (VVIR, DDD, and DDDR) in a randomised crossover fashion. SETTING: Pacemaker clinic at a university teaching hospital. PATIENTS: 15 patients with complete heart block, mean (SD) age 63 (10) years, and 12 patients with sick sinus syndrome, mean age 68 (9) years, implanted with DDDR pacemakers. MAIN OUTCOME MEASURES: Platelet activation was assessed by measuring the plasma concentrations of platelet factor 4 (PF4) and beta thromboglobulin using an enzyme linked immunosorbent assay (ELISA). Mean log plasma PF4 and beta thromboglobulin values were compared in paced patients during different pacing modes and with controls. RESULTS: Compared with controls, patients paced in DDDR, DDD, and VVIR modes had higher mean log plasma concentrations of PF4 (0.90 (0.32), 0.92 (0.29), and 1.12 (0.33) v 0.61 (0.29) log IU/ml, all p < 0.05, respectively) and beta thromboglobulin (1.55 (0.20), 1.59 (0.16), and 1.71 (0.18) v 1.40 (0.12) log IU/ml, all p < 0.05, respectively). In paced patients, VVIR pacing was associated with higher plasma concentrations of PF4 and beta thromboglobulin than either DDDR or DDD pacing (all p < 0.05). There was no significant difference in plasma PF4 and beta thromboglobulin between patients with complete heart block and sick sinus syndrome in the corresponding pacing mode. Holter monitoring showed no difference in mean pacing rate and occurrence of cardiac arrhythmias to account for the increased platelet activation during VVIR pacing. There was no relation between the percentage of ventricular pacing on Holter during DDDR, DDD, and VVIR modes and the log mean plasma concentrations of PF4 (r = 0.002, 0.001, and 0.001, respectively, all p > 0.05) and beta thromboglobulin (r = 0.007, 0.01, and 0.001, respectively, all p > 0.05). CONCLUSIONS: Single chamber ventricular pacing was associated with enhanced spontaneous systemic platelet activation compared with physiological dual chamber pacing. This was related to the loss of atrioventricular synchrony rather than to the underlying cause of bradycardia, lack of rate response, or coexisting arrhythmia. This abnormality may be associated with increased thromboembolism and was correctible by an appropriate pacing mode prescription and possibly antiplatelet treatment. PMID- 9404252 TI - Age dependent efficacy of implantable cardioverter-defibrillator treatment: observations in 450 patients over an 11 year period. AB - OBJECTIVE: To determine whether implantable cardioverter-defibrillator (ICD) treatment is beneficial in elderly patients with life threatening ventricular tachyarrhythmias. DESIGN: Since January 1984, ICDs were implanted in 450 patients to evaluate surgical risk, complications and mean survival in relation to patient age; 81 patients (18%) were < or = 50 years at the time of ICD implant, 254 patients (56%) were between 51 and 64 years, and the remaining 115 (26%) were > or = 65 years. Epicardial lead systems were implanted in 209 patients (46%), while transvenous lead systems were implanted in 241 (54%). RESULTS: 13 patients (3%) died perioperatively, more often after epicardial (11 of 209 patients, 5%) than after transvenous ICD implantation (one of 241 patients, < 1%) (p < 0.05). During a mean (SD) follow up of 28 (24) months (range < 1 to 114 months), 90 patients (20%) died. Of these, nine (2%) died from sudden arrhythmic death; five (1%) died suddenly, probably as a result of non-arrhythmic causes; 55 (12%) died from other cardiac causes (congestive heart failure, myocardial infarction); and 21 (5%) died from non-cardiac causes. The three, five, and seven year survival for arrhythmic mortality was 95% in patients < or = 50 years compared with a three year survival of 93% and a five and seven year survival of 91% in patients of 51 to 64 years, and a three, five, and seven year survival of 99% in patients > or = 65 years. 362 patients (80%) received ICD discharges (21 (43) shocks per patient), with a similar incidence among all three patient groups (< or = 50 years, 80%; 51 to 64 years, 81%; > or = 65 years, 79%). The time interval between ICD implant and the first ICD treatment was shorter in patients > or = 65 years (8 (8) months) than in patients between 51 and 64 years (11 (14) months) or < or = 50 years (11 (11) months) (p < 0.05). Survival time following first appropriate shock was 30 (24) months in patients < or = 50 years, 30 (26) months in patients of 51 to 64 years, and 19 (20) months in patients > or = 65 years. CONCLUSIONS: Elderly patients benefit from ICD treatment, and survive for a considerable time after the first treatment. Therefore, elderly patients should be considered candidates for ICD implantation if life threatening ventricular tachy-arrhythmias are present. PMID- 9404253 TI - Dispersion of ventricular repolarisation: a marker of ventricular arrhythmias in patients with previous myocardial infarction. AB - OBJECTIVE: To examine whether, in coronary patients after myocardial infarction, the dispersion of ventricular repolarisation measured through QT and JT intervals from a surface electrocardiogram could allow separation of those with ventricular tachyarrhythmias (VT) complicating their myocardial infarct from those without. DESIGN: A retrospective comparative study. SETTING: University hospital. PATIENTS: 39 patients with myocardial infarction complicated by VT, 300 patients after myocardial infarction without arrhythmic events, and 1000 normal subjects. The myocardial infarction groups were divided into anterior, inferior, and mixed locations. INTERVENTIONS: A computer algorithm examined an averaged cycle from a 10 second record of 15 simultaneous leads (12 lead ECG + Frank XYZ leads). After interactive editing, four intervals were computed: QTapex, JTapex, QTend, and JTend. For each interval, the dispersion was defined as the difference between the maximum and minimum values across the 15 leads. RESULTS: The mean values of all four dispersion indices were higher in patients with myocardial infarction than in normal subjects (p < 0.01). In the infarct groups, patients with VT had significantly greater mean and centile dispersion values than those without VT. For instance, the 97.5th centile value of QTend was 65 ms in normal individuals, 90 ms in infarct patients without arrhythmia, and 128 ms in those with VT; 70% of the infarct patients who developed serious ventricular arrhythmias had values exceeding the 97.5th centile of the normal group, while only 18% of the infarct patients without arrhythmia had dispersion values above this normal upper limit. Among the infarct patients, nearly half of those (18 of 39) with tachyarrhythmias had dispersion values that exceeded the 97.5th centile of those without arrhythmia. CONCLUSIONS: Dispersion of ventricular repolarisation may be a good non-invasive tool for discriminating coronary patients susceptible to VT from those who are at low risk. PMID- 9404254 TI - Long-term ventricular performance after intra-atrial correction of transposition: left ventricular filling is the major limitation. AB - OBJECTIVE: To establish the incidence of systolic and diastolic dysfunction of the right and left ventricle in a large cohort of patients after Mustard or Senning operations and to assess changes in the incidence on long term follow up. DESIGN: Postoperative case-control study using radionuclide ventriculography. Ejection fractions, peak filling rates, rapid filling periods and fractions, slow filling periods and fractions, and atrial contraction periods and fractions were studied. SETTING: Tertiary care centre, ambulatory and hospital inpatient care. PATIENTS: A convenience sample of 153 patients studied at median age of 6.9 years (median 4.4 years after surgery). In 99 cases another study was available at a median age of 15.3 years (median 13 years after surgery and 8.8 years after the first study). RESULTS: Respective incidences of dysfunction in the first and the second study were as follows: ejection fraction-right ventricle 7.8% and 8.1%, left ventricle 7.2% and 10.1%: peak filling rate-right ventricle 0% and 4.2%, left ventricle 14.3% and 29.5% (p < 0.05); rapid filling period-right ventricle 18.3% and 11.6%, left ventricle 30.2% and 30.5%; slow filling period-right ventricle 4.8% and 3.2%; left ventricle 11.9% and 23.2%; atrial contraction period-right ventricle 0.8% and 4.2%, left ventricle 15.1% and 26.3%; rapid filling fraction-right ventricle both 0%, left ventricle 82.5% and 79.0%; slow filling fraction-right ventricle 0.8% and 4.2%, left ventricle 37.3% and 30.5%; atrial contraction fraction-right ventricle both 0%, left ventricle 79.4% and 71.6%. CONCLUSIONS: The incidence of systolic ventricular dysfunction is 8% (right ventricle) and 10% (left ventricle) 13 years after surgery, without a significant increase over the eight year follow up. Diastolic filling is abnormal in up to 80% of patients and left ventricular peak filling rate deteriorates with time. PMID- 9404255 TI - Historical note: letter regarding heart surgery in England. PMID- 9404256 TI - Assessment of atrial septal defect morphology by transthoracic three dimensional echocardiography using standard grey scale and Doppler myocardial imaging techniques: comparison with magnetic resonance imaging and intraoperative findings. AB - OBJECTIVE: To determine whether transthoracic three dimensional echocardiography is an accurate non-invasive technique for defining the morphology of atrial septal defects (ASD). METHODS: In 34 patients with secundum ASD, mean (SD) age 20 (17) years (14 male, 20 female), the measurements obtained from three dimensional echocardiography were compared to those obtained from magnetic resonance imaging (MRI) or surgery. Three dimensional images were constructed to simulate the ASD view as seen by a surgeon. Measured variables were: maximum and minimum vertical and horizontal ASD dimension, and distances to inferior and superior vena cava, coronary sinus, and tricuspid valve. In each patient two ultrasound techniques were used to acquire three dimensional data: standard grey scale imaging (GSI) and Doppler myocardial imaging (DMI). RESULTS: Good correlation was found in maximum ASD dimension (both horizontal and vertical) between three dimensional echocardiography and both MRI (GSI r = 0.96, SEE = 0.05 cm; DMI r = 0.97, SEE = 0.04 cm) and surgery (GSI r = 0.92, SEE = 0.06 cm; DMI r = 0.95, SEE = 0.06 cm). The systematic error was similar for both three dimensional techniques when compared to both MRI (GSI = 0.40 cm (27%); DMI = 0.38 cm (25%)) and surgery (GSI = 0.50 cm (29%); DMI = 0.37 cm (22%)). A significant difference was found in both horizontal and vertical ASD dimension changes during the cardiac cycle. This change was inversely correlated with age. These findings were consistent for both DMI and GSI technique. In children (age < or = 17 years), the feasibility of detecting structures and undertaking measurements was similar for both echo techniques. However, in adult ASD patients (age > or = 18 years) this feasibility was higher for DMI than for GSI. CONCLUSIONS: Transthoracic three dimensional imaging using both GSI and DMI accurately displayed the varying morphology, dimensions, and spatial relations of ASD. However, DMI was a more effective technique than GSI in describing ASD morphology in adults. PMID- 9404258 TI - An association between arterial pulse pressure and variation in the fibrillin-1 gene. AB - OBJECTIVE: To investigate whether variation in the fibrillin-1 gene was associated with blood pressure in healthy middle aged men, as had been observed in patients with abdominal aortic aneurysm. DESIGN, SETTING, AND PATIENTS: Middle aged men (n = 245), aged 50 to 61 years, were recruited from one of the nine general practices participating in the second Northwick Park heart study. Blood samples were obtained for the preparation of genomic DNA and analysis of plasma variables. MAIN OUTCOME MEASURES: Systolic, diastolic, and pulse pressures; fibrillin-1 genotype characterised with a four allele variable tandem nucleotide repeat polymorphism in intron 28. RESULTS: In healthy middle aged men only three common genotypes were observed: 2-2 (frequency 54.1%), 2-3 (16%) and 2-4 (15%). The mean arterial systolic (and pulse) pressure varied according to fibrillin-1 genotype: 2-4 genotype, 126-3 (47.6) mm Hg; 2-2 genotype, 131.0 (51.3) mm Hg; and 2-3 genotype, 135.5 (54.2) mm Hg. The median pulse pressure was 50 mm Hg. Distribution of men around the median pulse pressure, according to genotype, showed a significant trend for patients of 2-4 genotype to have the lowest pulse pressures, those of 2-2 genotype to have intermediate pressures, and those of 2-3 genotype to have the highest pulse pressures (p = 0.003 for healthy men). CONCLUSIONS: There appears to be a significant association between fibrillin-1 genotype and arterial pulse pressure in men aged 50 to 61 years. PMID- 9404257 TI - Plasma concentrations of adrenomedullin correlate with the extent of pulmonary hypertension in patients with mitral stenosis. AB - OBJECTIVE: To examine the pathophysiological significance of adrenomedullin in the pulmonary circulation by investigating the relation between plasma concentrations of adrenomedullin and central haemodynamics in patients with mitral stenosis. METHODS: Plasma concentrations of adrenomedullin in blood samples obtained from the femoral vein, pulmonary artery, left atrium, and aorta were measured by a newly developed specific radio-immunoassay in 23 consecutive patients with mitral stenosis (16 females and seven males, aged 53 (10) years (mean (SD)) who were undergoing percutaneous mitral commissurotomy. RESULTS: Patients with mitral stenosis had higher concentrations of adrenomedullin than age matched normal controls (3.9 (0.3) v 2.5 (0.3) pmol/l, p < 0.001). There was a reduction in adrenomedullin concentrations between the pulmonary artery and the left atrium (3.8 (0.2) v 3.2 (0.4) pmol/l, p < 0.001). The venous concentrations of adrenomedullin correlated with mean pulmonary artery pressure (r = 0.65, p < 0.001), total pulmonary vascular resistance (r = 0.83, p < 0.0001), and pulmonary vascular resistance (r = 0.65, p < 0.001). Plasma concentrations of adrenomedullin did not change immediately after percutaneous mitral commissurotomy; however, they decreased significantly one week later. CONCLUSIONS: Plasma concentrations of adrenomedullin are increased in patients with mitral stenosis. This may help to attenuate the increased pulmonary arterial resistance in secondary pulmonary hypertension due to mitral stenosis. PMID- 9404259 TI - Heartstart Scotland: the use of paramedic skills in out of hospital resuscitation. AB - OBJECTIVE: To assess the frequency with which paramedic skills were used in out of hospital cardiac arrest and the effect of tracheal intubation on outcome. DESIGN: Retrospective analysis of ambulance service reports and hospital records. SETTING: Scottish Ambulance Service and hospitals admitting acute patients throughout Scotland. RESULTS: A total of 8651 out of hospital resuscitation attempts were recorded and tracheal intubation was attempted in 3427 (39.6%) arrests. One hundred and thirty six patients (3.7%) were intubated and 476 (9.1%) of the patients who were not intubated survived to hospital discharge (p < 0.001). Among the patients who were defibrillated the proportion intubated was highest in the patients who received the greatest number of shocks (p < 0.01). Among subjects receiving similar numbers of shocks survival rates were lower for intubated patients (p < 0.01). Patients with unwitnessed arrests were most frequently intubated and survival rates were lowest in this group. CONCLUSIONS: Patients who are intubated seem to have lower survival rates. This may however reflect the difficulty of the resuscitation attempt rather than the effects of intubation. The use of basic life support skills rapidly after cardiac arrest is associated with the best survival rates. PMID- 9404260 TI - Clinical competence in electrophysiological techniques. PMID- 9404261 TI - Junctional ectopic tachycardia in six paediatric patients. AB - The presenting features and treatment responses of six children with junctional ectopic tachycardia are evaluated. Two of the patients were siblings and both presented in early childhood with cardiopulmonary failure. The elder sibling died, the surviving sibling was controlled on a combination of amiodarone, digoxin, and sotalol. The remaining four patients presented in later childhood with tachycardia induced cardiomyopathy. Two of the patients were diagnosed incidentally and have normalised their myocardial function on sotalol therapy. The other two presented in congestive cardiac failure. Radiofrequency His bundle ablation and insertion of a permanent pacemaker to control the arrhythmia was undertaken in the elder of the two patients. The remaining patient has had marginal recovery of myocardial function on a combination of amiodarone and sotalol treatment. Improvement in myocardial function may take several months and is dependent on control of the tachycardia in some patients. Sotalol, when used as single or combination treatment, was partially successful in four cases in reducing heart rate. None of the patients reverted to sinus rhythm. PMID- 9404262 TI - Cardiac conduction abnormalities preceding transoesophageal echocardiographic evidence of perivalvar extension of infection in a case of Salmonella prosthetic valve endocarditis. AB - A 59 year old African-American man developed complete heart block in association with Salmonella enteritidis prosthetic valve endocarditis. Severe cardiac conduction abnormalities signalled the presence of perivalvar extension of infection before development of evidence of abscess by transoesophageal echocardiography. Cardiac conduction temporarily returned after debridement and aortic homograft placement. This case emphasises the value of electrocardiographic monitoring in the detection of perivalvar extension of infection complicating infective endocarditis, even in the era of sophisticated imaging modalities. PMID- 9404263 TI - Hibernating myocardium caused by isolated, radiation induced left main stem coronary artery stenosis. AB - A 45 year old man presented with a five week history of worsening exertional dyspnoea and orthopnoea. He had also noted mild, bilateral ankle swelling. The patient had been diagnosed with stage III Hodgkin's lymphoma in 1968 at the age of 21. During the same year he underwent total nodal irradiation followed by chemotherapy in 1971. He had remained entirely asymptomatic over the course of the next 24 years with no evidence of relapse. Cardiac catheterisation undertaken soon after admission revealed a tight left main stem stenosis with a left dominant system. Left ventriculogram showed severe, global hypokinesia, and raised left ventricular end diastolic pressure (22 mm Hg). Urgent coronary artery bypass graft surgery was carried out. He made an uncomplicated recovery and his condition improved sufficiently to allow discharge eight days following the procedure. His heart failure slowly resolved and repeat transthoracic echocardiogram performed six months after surgery showed an unequivocal improvement in left ventricular function. Left ventricular ejection fraction continued to improve and increased from 23% at two months to 42% at two years. He currently remains entirely asymptomatic off all medication. PMID- 9404264 TI - Atrial fibrillation begets trouble. PMID- 9404265 TI - Increase in hospital admission rates for heart failure in The Netherlands, 1980 1993. PMID- 9404266 TI - Coronary patients need cardiologists. PMID- 9404267 TI - Follicular growth and ovarian dynamics in mammals. AB - General characteristics of female reproductive activity, such as seasonality, cyclicity and triggering of ovulation differ widely among mammals, but common mechanisms underlie ovarian function. In all mammals, follicles begin to grow from a pool of primordial follicles constituted early in life, continuously throughout the life of the female. Follicular development involves two phases. In a first phase (basal follicular growth), follicles grow slowly and follicular growth rate is tightly related to proliferation of granulosa cells. Basal follicular growth is mainly under the control of growth factors of paracrine origin. In these follicles, FSH may exert an indirect mitogenic effect on granulosa cells by enhancing expression of growth factors or growth factor receptors. In a second phase (terminal follicular growth), follicular growth is rapid and occurs by enlargement of the antrum. In addition, it is accompanied by important changes in differentiation of follicular cells. Terminal follicular development is strictly dependent on gonadotrophins. FSH plays determinant roles in enhancing granulosa cell differentiation and survival. These actions are mediated or modulated in an important way by paracrine factors, particularly steroids and growth factors. LH stimulates steroidogenesis in theca cells and sustains terminal maturation of granulosa cells in preovulatory follicles. Follicular growth, atresia and ovulation are accompanied by important tissue remodelling processes, which are under the fine control of proteinases and inhibitors of proteinases. In particular matrix metalloproteinases and their inhibitors are probably involved in the control of rapid terminal follicular growth and regression of atretic follicles as well as in follicular rupture at ovulation. PMID- 9404268 TI - Changes in the number of follicles and of oocytes in ovaries of prepubertal, peripubertal and mature bitches. AB - Ovaries were collected from prepubertal (< 6 months of age, n = 4 ovaries), peripubertal (6 to 10 months of age, n = 12 ovaries) and mature (> 10 months, n = 12 ovaries) bitches after routine ovariohysterectomies and fixed in formalin. Ovaries were bisected, embedded in paraffin wax and 20 serial sections were made at intervals of 10 microns. Sections were stained with haematoxylin and eosin to examine follicles and oocytes in a cross-section of cortex of known size. Counts were made on all sections, resulting in examination of the entire cortical area present in the sections. Oocytes were counted and classified as nucleate or anucleate. Follicles were counted and classified as large (> 100 microns in diameter) or small (< 100 microns in diameter), containing one oocyte (monovular), more than one oocyte (polyovular) or no oocytes (anovular). It was concluded that at first oestrus there was an increase (P < 0.05) per section in number of total oocytes and small monovular follicles and a significant increase (P < 0.05) in the number of nucleated oocytes in monovular follicles, suggesting that oogenesis or folliculogenesis is still occurring at this age. At pre- and peripubertal ages polyovular follicles were found which persist into maturity. There was no difference in numbers of anovular follicles and total number of polyovular follicles among different age groups. PMID- 9404269 TI - New methods for gamete rescue from gonads of nondomestic felids. AB - Methods for rescuing oocytes and spermatozoa post mortem are described, which were adapted from domestic cat as a model. Ovaries were mechanically processed for large-scale recovery of oocytes. Numbers of intact cumulus-oocyte complexes (COCs) per animal and of preantral follicles per ovary were 18 +/- 2 and 2892 +/- 665 in domestic cats, respectively. Similar results were obtained from 13 individuals of 6 nondomestic felids: 16 +/- 2 COCs and 1867 +/- 1144 follicles. Preantral follicles were cultured for at least 7 days. Intact COCs were maturated for 24 h and fertilized in vitro with homologous or heterologous (from domestic cat) spermatozoa. Spermatozoa were collected from caudae epididymides (n = 11; five nondomestic species) and cryopreserved (n = 8) using a programmed freezer. The reproductive competence of oocytes collected post mortem was demonstrated by development to embryos (> or = 8 cells) in vitro. Spermatogenic efficiency of males was assessed by flow cytometric analysis of mitotic and meiotic testicular cells as well as by estimation of testosterone concentration in testes. The results demonstrate the possibility of retrospective assessment of male and female reproductive capacity. In conclusion, the described methods could be a useful part of gamete rescue programmes for endangered felids. PMID- 9404270 TI - LH release, induction of oestrus and fertile ovulations in response to pulsatile administration of GnRH to anoestrous dogs. AB - In one study in 33 bitches, in which each of seven doses of GnRH were injected into six anoestrous bitches each, doses of > or = 400 ng kg-1 released LH consistently, and doses of < or = 200 ng kg-1 did not. In a second study, GnRH was administered in pulses of 15-500 ng kg-1 every 90 min for 7-9 days to 36 anoestrous bitches, four prepubertal bitches and three luteal phase bitches. In 36 anoestrous bitches, GnRH pulses resulted in pro-oestrus, oestrus, ovulation and pregnancy in 26, 20, 16 and 12 bitches, respectively. Mean time for pro oestrus onset, oestrus, ovulatory LH surge and parturition were 5.1 +/- 0.4, 11.2 +/- 0.7, 14.0 +/- 9.0, and 77.3 +/- 1.0 days, respectively, from start of treatment. In response to GnRH, pro-oestrus and pregnancy occurred in two of four prepubertal bitches, and pro-oestrus but not ovulation occurred in one of three luteal phase bitches. Efficacy in anoestrous bitches was dose dependent with high doses of > 280 ng kg-1 (n = 12) versus intermediate 85-270 ng kg-1 (n = 12) and low < 85 ng kg-1 doses (n = 12) resulting in a higher incidence of pro-oestrus (100 versus 80 and 33%), oestrus (92 versus 50 and 25%), ovulation (84 versus 42 and 8%) and pregnancy (58 versus 33 and 8%). With few exceptions, failed or weak responses were related to inadequate LH release, as measured in frequently collected samples following successive pulses on selected days of treatment; vaginal cornification, oestrous behaviour, and LH response were similar in oestrus without ovulation and fertile oestrus except for the lack of an ovulatory LH surge. PMID- 9404271 TI - Dopamine agonistic effects as opposed to prolactin concentrations in plasma as the influencing factor on the duration of anoestrus in bitches. AB - The duration of the interoestrous interval and plasma concentrations of prolactin were determined in seven bitches treated with 0.1 mg metergoline kg-1 body mass orally, twice a day during anoestrus to determine whether shortening of the interoestrous interval in dogs is due to suppression of prolactin secretion. The results were compared with those of a previous experiment in which four bitches were treated with 20 micrograms bromocriptine kg-1 body mass orally during the luteal period and anoestrus. The interoestrous interval of the dogs treated with metergoline, 240 +/- 67 days, was similar to the interoestrous interval before treatment, 225 +/- 54 days. However in the dogs treated with bromocriptine, the interoestrous interval was shortened to 123 +/- 46 days. The average of the means of the plasma concentrations of prolactin during metergoline treatment, 1.5 +/- 0.5 micrograms l-1, was lower (P = 0.03) than the average of the means of the plasma concentrations of prolactin during the 2 weeks before treatment, 3.6 +/- 2.3 micrograms l-1, but did not differ from the average of the means of the plasma concentrations of prolactin, 2.1 +/- 0.5 micrograms l-1, of the dogs treated with bromocriptine during anoestrus. As the plasma concentrations of prolactin were similar during metergoline and bromocriptine treatment, and the anoestrous period was shortened during bromocriptine treatment, but not during metergoline treatment, these results suggest that the induction of oestrus by dopamine agonists is not initiated by suppression of prolactin secretion, but by other direct or indirect dopaminergic effects. PMID- 9404272 TI - Termination of dioestrus and induction of oestrus in dioestrous nonpregnant bitches by the prolactin antagonist cabergoline. AB - The effects of treatment with a dopamine agonist from day 30 after the LH surge of a nonpregnant oestrous cycle were analysed in five bitches previously demonstrated to be fertile. Five untreated nonpregnant bitches were studied as controls. Cabergoline was given (5 micrograms kg-1 body weight) daily from day 30 after the LH peak until +/- 2 days after the first sign of pro-oestrus. Cabergoline decreased progesterone concentrations to < 1 ng ml-1 within 15-20 days. Four of the five treated bitches came into pro-oestrus after a mean of 29.75 +/- 5 days of treatment, at 66.5 +/- 7.1 days after the previous LH surges. The previous and following interoestrous intervals in the treated and responding animals were 216 +/- 6 and 66.5 +/- 7.1 days, respectively, and were significantly different (P < 0.05). In untreated animals, interoestrous intervals were longer (P < 0.05) than in treated animals, and not different from previous interoestrous intervals. The duration of pro-oestrus and oestrus was normal in the induced oestrous cycle compared with previous or untreated cycles in both groups. Variations in progesterone in the induced cycles were not different from those in control cycles. Prolactin concentrations were lower (P < 0.05) in treated animals compared with untreated animals. All bitches mated. However, none of the treated bitches became pregnant at the induced oestrous cycle. Lack of fertility was presumably due to the inability of the uterus to receive the embryos and was related to insufficient uterine involution and regeneration. Nevertheless, this study demonstrates that it is possible to induce oestrus not only in anoestrus but also in dioestrous nonpregnant bitches using a D2 dopamine agonist. PMID- 9404273 TI - In vitro and in vivo development of embryos produced by in vitro maturation and in vitro fertilization of cat oocytes. AB - Cumulus-oocyte complexes of domestic cats were classified by morphology of ooplasm (A = good, B = fair, C = poor) and cultured for 24 h in TCM 199 with gonadotrophins (eCG, FSH, hCG or FSH/hCG). More of type A oocytes (52%) underwent in vitro maturation (IVM) than of type B (41%) or type C (17%). The gonadotrophin source did not affect frequency of IVM of type A (50-53%) or type B (38-44%) oocytes, but IVM of type C oocytes in hCG or FSH/hCG (27%/19%) was about double that in eCG or FSH alone (13%/10%). After IVF, frequency of cleavage for type A (54%), B (41%) and C (26%) oocytes was similar to the IVM frequency of the equivalent type. After 7 days, development to the morula (M) stage in vitro was similar among types (47-58%); however, higher percentages of type A and B oocytes developed to blastocysts (Bl), 31% and 29%, respectively, than of type C (15%). After transfer of day 5 (n = 70) or 6 (n = 32) M and Bl to day 4 or 5 recipients in trial 1 (n = 4) and 2 (n = 3), respectively, the three recipients in trial 2 gave birth to four live kittens. Development in vitro to M of IVM/IVF embryos frozen in propanediol plus sucrose during early cleavage was similar (64-69%) to that of cohort controls (64%), but Bl formation was reduced (13-17% versus 32%). Damage to the zona pellucida after plunging into liquid nitrogen at -30 degrees C was higher (11%) than that of the embryos cooled at 10 degrees C min-1 from -30 degrees C to -150 degrees C before storage (2%). PMID- 9404274 TI - Effect of preovulatory endocrine events upon maturation of oocytes of domestic bitches. AB - The effect of preovulatory endocrine changes upon oocyte maturation was investigated in a combined in vivo and in vitro study. A preliminary study was performed to investigate the effect of endocrine influences in vivo on subsequent oocyte maturation in vitro. A hemi-ovariectomy was performed in four bitches during pro-oestrus at the first detected rise of plasma progesterone (mean progesterone 2.36 +/- 0.58 ng ml-1) and a second performed during oestrus immediately before ovulation, 1-4 days later (mean progesterone 4.96 +/- 0.75 ng ml-1). The mean numbers of high quality (grade 1) oocytes with identifiable nuclear material collected per pro-oestrous ovary was 15.5 +/- 9.88, and per oestrous ovary was 31.75 +/- 23.76. All of the grade 1 oocytes harvested were cultured for 96 h, after which 37% and 19% of pro-oestrous oocytes had matured to germinal vesicle breakdown (GVBD) and metaphase I/anaphase I/metaphase II (MI/AI/MII), respectively, whereas 41% and 11% of oestrous oocytes had matured to GVBD and MI/AI/MII, respectively. There was no significant difference in maturation between the two groups of oocytes. In a second study, high quality grade 1 oocytes harvested from bitches in the late luteal or anoestrous stage of the oestrous cycle were cultured in medium supplemented with either 1 microgram oestradiol ml-1 (64 oocytes), 1 microgram progesterone ml-1 (52 oocytes), a combination of oestradiol and progesterone (50 oocytes), or no supplementation (55 oocytes). The mean numbers of high quality (grade 1) oocytes with identifiable nuclear material collected per luteal ovary was 9.71 +/- 7.96, and per anoestrous ovary was 8.33 +/- 5.61. The number of oocytes obtained per bitch was highly variable, and may have been affected by the stage of oestrous cycle. After 48 h, maturation to GVBD was 56, 33, 50 and 50%, and to MI/AI/MII was 13, 17, 20 and 4% for each treatment respectively, and after 96 h was 45, 59, 63 and 63% and 10, 0, 4 and 0%, respectively. There was no significant difference in maturation between the four groups. The hormonal environment in vivo did not affect subsequent in vitro maturation of high quality oocytes. Similarly, culture in steroid hormone supplemented medium did not improve maturation in vitro of high quality oocytes. The effects upon oocytes from specific preovulatory follicles were not studied. PMID- 9404275 TI - Effects of slow and ultrarapid freezing on morphology and resumption of meiosis in immature cat oocytes. AB - This study examined the ability of immature cat oocytes to survive after cryopreservation by evaluating their subsequent development following maturation in vitro. The effect of slow and ultrarapid freezing using one of two cryoprotectants dimethylsulphoxide (DMSO) or 1,2-propanediol (PROH) at different concentrations (1.5 or 3.0 mol-1) and the slow freezing with the cryoprotectant ethylene glycol (EG 1.5 mol l-1 and 3.0 mol l-1) were tested. Morphology, resumption of meiosis and metaphase II rates of oocytes were recorded after thawing. Freshly collected oocytes were used as controls. Results indicate that immature cat oocytes can survive, resume meiosis and achieve metaphase II in vitro after freezing. The highest rates of resumption of meiosis and metaphase II were achieved with slow freezing and 1.5 mol DMSO or EG l-1 (DMSO: 47.4%, 18/38 and 23.7%, 9/38 and EG: 52%, 13/25 and 20%, 5/25, respectively). The ultrarapid procedure did not result in resumption of meiosis in vitro, despite intact morphology of the oocytes after thawing. These results suggest that morphology of oocytes after freezing and thawing has no predictive value for their ability to resume meiosis. PMID- 9404276 TI - Effects of cooling, freezing and glycerol on penetration of oocytes by spermatozoa in dogs. AB - A homologous zona penetration assay was used to evaluate dog spermatozoa after selected treatments and associated gamete interaction with sperm characteristics. Canine semen, diluted in egg-yolk extender, was treated to compare samples quickly cooled to 0 degree C (after 0.5 and 3 h), slowly cooled to 0 degree C after 3 h or held for 3 h at room temperature. In a second study, slowly cooled spermatozoa with or without the addition of 4% glycerol were studied. After each treatment, spermatozoa were incubated with canine oocytes for 18 h, stained with Hoechst and examined for bound sperm heads. After 3 h, cells that had been cooled quickly demonstrated lower numbers of spermatozoa per ovum than fresh or slowly cooled treatments (2.4 +/- 0.6 versus 5.2 +/- 1.1 and 3.6 +/- 0.9 sperm per oocyte, respectively) and coincided with a reduction in acrosomal integrity and motility (58.6% fast cool 3 h versus 83.1% fresh). Addition of glycerol effected no change in acrosomal status or motility, but resulted in a decline in spermatozoa bound per oocyte to 1.5 +/- 0.3. While freezing caused a further decline in motility, acrosomal status and oocyte penetration, there was no difference between freezing with and without seeding (0.3 +/- 0.08 versus 0.2 +/- 0.04 spermatozoa per ovum). Evaluation of sperm penetration of homologous oocytes provides new insights into the effects of cooling, seeding and glycerol on canine spermatozoa. PMID- 9404277 TI - Semen quality after thawing: correlation with fertility and fresh semen quality in dogs. AB - Fifty-four semen samples from five dogs were evaluated both, fresh and after thawing. Some of these semen samples were mixed with autologous prostatic fluid after thawing and used to inseminate each of nine bitches 4-7 times intravaginally. All bitches conceived and the mean number (+/- SD) of conceptuses, number of corpora lutea and ratio between conceptuses and corpora lutea (implantation rate) were 5.7 +/- 2.8, 9.4 +/- 1.1 and 0.63 +/- 0.34, respectively. The mean incidence of normal sperm morphology and progressively motile spermatozoa for all semen samples were 71.5 +/- 13.5% and 74.4 +/- 7.1%, respectively, in fresh semen and 52.0 +/- 18.5% and 53.4 +/- 12.6% in frozen thawed semen. Extension rate was 1:3 for all semen samples and the mean sperm concentration after thawing was 12.08 +/- 6.66 x 10(7) ml-1. The only semen quality variables after thawing that were correlated with implantation rate were the number of spermatozoa inseminated on day-2 and number of progressively motile spermatozoa inseminated on day-2 (where day 0 is the day of onset of dioestrus as determined by cytology) (Spearman's rank correlation coefficient > 0.7, n = 9, P < 0.05). This study suggests that it is essential that frozen-thawed semen is inseminated on day-2 and that an insemination dose of 10-11 x 10(7) progressively motile frozen-thawed spermatozoa is adequate to achieve a mean implantation rate of 75% or higher. The incidence of either proximal or distal cytoplasmic droplets in fresh semen was negatively correlated with motility after thawing in three of five dogs (Spearman's rank correlation < -0.5, n = 6-17, P < 0.05). Neither the percentage spermatozoa with normal morphology in fresh semen nor the percentage progressively motile spermatozoa in fresh semen nor the concentration of spermatozoa after thawing were correlated with motility after thawing. Fresh semen quality, with the exception of the incidence of retained cytoplasmic droplets, has little value in predicting the progressive motility after thawing in frozen dog semen. PMID- 9404278 TI - Effect of pentoxifylline on motility and longevity of fresh and thawed dog spermatozoa. AB - The effect of pentoxifylline on fresh and frozen-thawed dog spermatozoa was evaluated. In Expt 1, 0.0036 mol l-1 or 0.0072 mol l-1 pentoxifylline was added to fresh dog spermatozoa. Semen was incubated at 25 degrees C or 39 degrees C. In Expt 2, pentoxifylline was added either before freezing or in the process of thawing at a final concentration of 0.0072 mol l-1. Addition of pentoxifylline significantly increased the percentage of progressively motile spermatozoa in fresh semen. The most beneficial effect of pentoxifylline on frozen-thawed semen was observed (P < 0.001) when it was added to semen in the process of thawing. PMID- 9404279 TI - Effect of progestogens and androgens upon spermatogenesis and steroidogenesis in dogs. AB - Two different progestogens, mixed testosterone esters, and a combination of progestogen and androgens were evaluated for their effect on semen quality and peripheral plasma testosterone and LH concentration in dogs. Megestrol acetate (2 mg kg-1) given orally for 7 days, and medroxyprogesterone acetate (10 mg kg-1) given subcutaneously produced no change in semen quality compared with that of control dogs. Higher doses of megestrol acetate (4 mg kg-1) produced minor secondary sperm abnormalities, whereas 20 mg medroxyprogesterone acetate kg-1 produced a rapid and significant decrease in sperm motility, morphology and output. The effect of progestogen was probably mediated by action upon the epididymis, since semen quality declined rapidly, morphological changes were the result of more secondary sperm abnormalities, and there was no suppression of plasma LH concentration. Mixed testosterone esters (5 mg kg-1) produced a significant decline in semen quality, which occurred 3 weeks after treatment and persisted for 3 months. There was an increase in the number of primary sperm abnormalities, and it was thought that the effect was probably related to suppression of gonadotrophin resulting in an effect on spermatogenesis. The combination of mixed testosterone esters (5 mg kg-1) and medroxyprogesterone acetate (20 mg kg-1) produced a rapid and profound decrease in semen quality. It was postulated that the effect of this combination of treatment on semen quality was mediated directly by the progestogen upon the epididymal phase of development, and the androgen causing suppression of gonadotrophins. Indeed a reduction in the plasma LH concentration was noted, and both primary and secondary sperm abnormalities were present. The dog appears to differ from other species in the sensitivity of the pituitary-hypothalamus to progestogen feedback. Gonadotrophin suppression does not occur even when high doses of progestogens are used and there are no significant effects on libido. However, combinations of progestogens and androgens may provide a clinically useful method of reversible contraception in the dog. PMID- 9404280 TI - Effect of finasteride (Proscar MSD) on seminal composition, prostate function and fertility in male dogs. AB - This study reports the long-term effects and reversibility of the administration of Finasteride, a 5 alpha-reductase inhibitor, on the prostate, prostatic fluid composition and volume, sperm output and characteristic, and fertility of adult beagle dogs treated for 21 weeks at a dose of 1 mg kg-1 body weight. Finasteride was not associated with any side effects. After 5 to 15 weeks of treatment, it induced a marked decrease in the size of the prostate and a fall in its secretions and in the production of spermatozoa. Sperm concentration increased and was inversely correlated with the decrease in the volume of prostatic secretions. At maximum effect, the calculated prostate volume was reduced to 30% of the initial value, and together with the reduced prostatic secretion, no ejaculate could be collected despite normal behaviour. These effects were reversible in 6 to 8 weeks after the cessation of treatment. Matings at 20 to 22 weeks after the start of treatment were fertile, demonstrating the absence of long-term effects of this treatment on male fertility. As fertility was not impaired and prostatic benign hyperplasia successfully regressed. Finasteride treatment in this preliminary study appears to be an interesting alternative therapy in valuable breeding dogs with benign prostatic hyperplasia. PMID- 9404281 TI - Luteal and placental characteristics of carnivore gestation: expression of genes for luteotrophic receptors and steroidogenic enzymes. AB - Experiments were carried out to investigate the abundance of mRNA for luteotrophic receptors and steroidogenic elements in the ovaries and corpora lutea of mink during the embryonic diapause, peri-implantation and postimplantation pregnancy. The second aim was to determine whether the mink placenta synthesized progesterone. Homologous cDNA probes for the mink LH and prolactin receptors were generated by the polymerase chain reaction. Heterologous cDNA probes for steroidogenic acute regulatory protein (StAR), cytochrome P450 side chain cleavage (P450scc) and 3 beta-hydroxysteroid dehydrogenase-delta 4 delta 5 isomerase (3 beta HSD) were also used. The abundance of mRNA encoding the prolactin receptor was low during the period of embryonic diapause and increased concurrent with circulating progesterone. The abundance of LH receptor message reached peak values during the peri-implantation period followed by maintenance of a steady-state after implantation. The abundance of StAR and P450scc messages appeared not to vary during gestation, while that for 3 beta HSD was correlated with changes in circulating progesterone. There was no evidence of 3 beta HSD activity or transcripts in the placenta. These results indicate that prolactin and LH are necessary for activation of the corpus luteum during the period of embryonic diapause, and for its maintenance during postimplantation gestation. The mink placenta does not synthesize progesterone. PMID- 9404282 TI - Immunocytochemical localization of oestrogen and progesterone receptors in the uterus of the normal bitch during oestrus and metoestrus. AB - Frozen uterine biopsy samples obtained from eight bitches during oestrus and metoestrus were used to develop immunocytochemical methods for identifying steroid receptors. The use of anti-steroid receptor monoclonal antibodies DAKO ID5 and NCL-PGR resulted in reliable detection of oestrogen and progesterone receptors, respectively. The distribution and heterogeneity of both oestrogen receptor and progesterone receptor expression varied during the oestrous cycle. During oestrus, steroid receptor expression was greatest as indicated by a distinct and homogeneous distribution of nuclear staining within cells of the endometrial glandular and luminal epithelium and myometrium. In contrast, during metoestrus there was an overall reduction in both the number and intensity of staining of nuclei. Distribution and intensity of nuclear staining was heterogeneous. This reduction was obvious within the luminal epithelial cells where nuclear staining was negligible compared with that of the glandular epithelial cells. This method could be used for further receptor studies, particularly in those diseases mediated by steroid hormones such as cystic endometrial hyperplasia and mammary neoplasia. PMID- 9404283 TI - Incidence of spontaneous ovulation in young, group-housed cats based on serum and faecal concentrations of progesterone. AB - Cats are considered to be reflex ovulators that exhibit a luteal phase (pregnancy or pseudopregnancy) only after copulatory stimulation. In a group-housed colony of 15 1-year-old domestic queens, 23 noncopulatory, spontaneous ovulations were observed in 87% of the queens over 4.5 months based upon the detection of increased concentrations of progesterone in faeces, serum, or both. Luteal phases were detectable for periods of 3 to 5 weeks with peak progesterone concentrations averaging 21 +/- 1 ng ml-1 in serum and 1874 +/- 281 ng g-1 in faeces. Individual cats exhibited from 0 to 3 spontaneous ovulations and subsequent pseudopregnancies each. A male was added to a separate cage within the room during the last 1.5 months of the study. The incidence of ovulation per 10 day period ranged from 0% to 22% before introduction of the male and from 33% to 57% immediately after introduction of the male, suggesting a potential noncopulatory influence of the male on the incidence of spontaneous ovulation in young, group housed cats. PMID- 9404284 TI - Dynamics of haemostasis during the oestrous cycle and pregnancy in bitches. AB - The blood coagulation status was studied in 31 bitches of different breeds during 33 oestrous cycles and during nine pregnancies. Two other bitches were ovariohysterectomized and received subcutaneous injections of oestradiol benzoate for 7 consecutive days. Blood samples were taken in early and late follicular phases, at ovulation, at day 1 after the end of oestrus as determined by cytology, at days 30, 60, 90 and 120 of metoestrus and in anoestrus. The samples were analysed for the concentrations of fibrinogen, fibrin(ogen) degradation products, as well as for the prothrombin time, the activated partial thromboplastin time, the antithrombin III activity, the number of platelets and the haematocrit. In other blood plasma samples the concentrations of oestradiol and progesterone were measured. In the two bitches that were ovariohysterectomized and received subcutaneous injections of oestradiol benzoate for 7 consecutive days, the coagulation parameters and hormones were examined in blood samples collected at appropriate terms and time intervals as in intact dogs. The significantly increased concentrations of fibrinogen and fibrin(ogen) degradation products, the large number of platelets and the decreased antithrombin III activity observed during the luteal phase of the nonpregnant and pregnant bitches are attributed to direct or indirect effects of the high peripheral progesterone concentrations. In the mid-luteal phase (day 30) this activation was more distinct during pregnancy than in the nonpregnant dogs presumably owing to additional effects of local processes in the uteroplacental area. Influences of high concentrations of oestradiol were not observed either during the follicular phase of the intact bitches or after oestradiol benzoate administration in the ovariohysterectomized dogs. PMID- 9404285 TI - Prolactin profiles of pregnant, lactating and non-mated female mink (Mustela vison). AB - This study was part of an experiment on energy metabolism in pregnant and lactating female mink (Mustela vison). Ten mated and three non-mated female mink were kept in metabolic cages in the laboratory from immediately after mating until the kits were three to four weeks old. Consecutive energy balance experiments with periods each of one week duration, including a 22 h respiration experiment (only three with females that were not mated) were performed, and weekly blood samples were collected for determination of plasma concentrations of thyroid hormones, prolactin, insulin and glucose. Prolactin profiles of pregnant and lactating females had a biphasic pattern: there was an increase in April, a decline immediately before parturition, and peak values were recorded in early May, usually when the females were in the first week of lactation. In females that were not mated plasma concentrations of prolactin did not rise above basal concentration. These females also exhibited a delayed spring moult, and had a lower feed intake. In addition, plasma profiles of thyroid hormones of mated and non-mated animals were different; concentrations of thyroid hormones decreased in April to early May, but the decrease started earlier and was most pronounced in pregnant female mink. These data indicate that prolactin secretion in female mink is regulated by photoperiod, but that other endocrinological events during pregnancy may also be involved. PMID- 9404286 TI - Secretion patterns of plasma prolactin and progesterone in pregnant compared with nonpregnant dioestrous beagle bitches. AB - The plasma concentration of prolactin has been poorly described in dogs. Published data are still controversial. In this study, plasma prolactin and progesterone concentration patterns were determined in groups of ten pregnant and ten nonpregnant beagle bitches. Blood samples were collected daily between 08:30 h and 09:30 h from day-15 to day 135 after the LH peak. Plasma concentrations of prolactin increased from day 25 after the LH surge in the pregnant animals to a peak value on day 65, the mean day of parturition. After parturition, they fell for 24-48 h, and then increased again and remained high during lactation. Variations observed in lactation were great and probably due to suckling. In the nonpregnant bitches, plasma prolactin remained constant throughout the observation period. An increase was observed at about day 70, but was not significant. Plasma prolactin patterns differed greatly, depending on whether the bitch was pregnant or not, even though the luteal phases had the same apparent duration in both cases. No significant difference was observed between progesterone concentrations of pregnant and nonpregnant bitches, even if plasma progesterone was higher from day 40 in pregnant animals. The luteal period ended abruptly in pregnant bitches on day 64, that is 1 day before parturition, and was more progressive in nonpregnant animals. Significantly higher prolactin secretion in pregnancy suggests specific luteal regulation, whereas the lower concentrations observed in nonpregnant dioestrous bitches make its role in nonpregnant luteal phases questionable. PMID- 9404287 TI - Progestogen concentration and ionic composition of the mammary secretion of periparturient bitches. AB - Mammary secretions were collected from nine bitches of various breeds during the periparturient period. Initial mean progestogen concentrations were high and started to decline from 5 days before parturition, in a similar way to changes in plasma progesterone concentration. Values were basal by 3-4 days after parturition. Marked individual and day variations prevented this method from reliably indicating the time of impending parturition. Calcium concentrations in milk increased from 3-5 days after parturition, whereas a decline in magnesium concentration was noted from day 1 postpartum. There was an increase in the Ca:Mg ratio from 1 day after parturition, although this was not significant until day 3, and was therefore not useful for predicting impending parturition. There were no significant changes in sodium concentration in milk. PMID- 9404288 TI - Characteristics of an oviductal glycoprotein and its potential role in fertility control. AB - At the time of ovulation the lining epithelium of the mammalian oviduct consists of columnar ciliated and secretory cells. These mature cells are dependent on ovarian steroids in carnivores. Oestradiol induces differentiation of these cells and maintains their mature functional state, and progesterone induces dedifferentiation. The secretory cells synthesize and secrete an oestrogen dependent high molecular weight glycoprotein. The cDNAs encoding oviductal glycoproteins from several species have been sequenced and show high similarity. The human cDNA hybridized with a single message on northern blots of total oviduct RNA obtained from oestradiol-treated cats (about 2.3 kb) and dogs (about 2.1 kb). This glycoprotein is the major nonserum protein present in the oviductal lumen at the time of ovulation, fertilization and early embryonic development. The glycoproteins associate with the zona pellucida of oviductal eggs in all species studied to date. Recent studies suggest that the bovine glycoprotein facilitates sperm capacitation and significantly increases the ability of bovine spermatozoa to fertilize bovine oocytes in vitro, that the hamster glycoprotein increases the sperm penetration rate of the zona pellucida by three times and that the human glycoprotein increases sperm binding to the zona pellucida by three times. All of the evidence for a biological function for this glycoprotein is derived from studies performed in several different species at reproductive stages before fertilization. The biological actions of this glycoprotein suggest a potential role for the glycoprotein in fertility control. Specifically, purified or recombinant glycoprotein may improve success in IVF procedures by enhancing binding of spermatozoa to the zona pellucida and improving fertilization rates. The glycoprotein may also be a potential immunocontraceptive target since antibodies generated against the oviductal glycoprotein may prevent fertilization by preventing binding of spermatozoa to the zona pellucida. PMID- 9404289 TI - Comparison of long-term effects of ovariectomy versus ovariohysterectomy in bitches. AB - Although ovariectomy is less invasive and less time-consuming than ovariohysterectomy, most surgical textbooks recommend ovariohysterectomy for routine neutering of bitches. This advice is probably based on concerns about the development of uterine disease after ovariectomy. However, there is no evidence that conditions such as cystic endometrial hyperplasia (CEH)-endometritis develop in the ovariectomized bitch, unless progestagens are administered. The purpose of this study was therefore to compare the long-term effects of ovariectomy and ovariohysterectomy, including the incidence of urinary incontinence. Questionnaires were sent to 264 owners of bitches, in which ovariectomy (126) or ovariohysterectomy (138) had been performed as a routine neutering procedure 8-11 years earlier. Complete data were available for 69 bitches of the ovariectomy group and for 66 bitches from the ovariohysterectomy group. There were no indications that endometritis had developed in bitches of the ovariectomy group. None of the bitches was sexually attractive to male dogs after neutering. The occurrence of a clear to white vaginal discharge was reported in two bitches of each group, but none of these four bitches appeared to be ill during the periods when the discharge was present. Furthermore, with the exception of urinary incontinence, no problems were reported that could be related to the surgical neutering. Six of the ovariectomized bitches and nine of the ovariohysterectomized bitches eventually developed urinary incontinence. Of these 15 bitches (11%), 12 weighed more than 20 kg. Bouvier des Flandres bitches were at a higher risk of developing urinary incontinence than were those of the other breeds. The possibility that the urinary incontinence was due at least in part to other conditions must be considered, since eight of the bitches were 9 years or older before urinary incontinence occurred and seven of the incontinent bitches also had polyuria or polydipsia. There were no significant differences in the incidence of urogenital problems listed above between the bitches of the ovariectomy and ovariohysterectomy group. It is hypothesized that a uterine disease such as CEH-endometritis cannot develop after complete ovariectomy, unless progestagens are administered. The results of this study indicate that ovariectomy does not increase the risk of CEH-endometritis or other complications in comparison with ovariohysterectomy. It is concluded that there is no indication for removing the uterus during routine neutering in healthy bitches. On the contrary, ovariectomy should be considered the procedure of choice. PMID- 9404290 TI - Clinical use of dexamethasone for termination of unwanted pregnancy in dogs. AB - Dexamethasone was administered orally for 7.5 or 9.5 days to 80 pregnant bitches to terminate unwanted pregnancy at an outpatient clinic. Treatment was initiated at day 30-35 (n = 74) or day 45-50 (n = 6) of confirmed pregnancy using one of two dosages. In 62 bitches, dexamethasone was administered twice a day for 7.5 days increasing from 0.1 to 0.2 mg kg-1 over the first 2 days of administration and decreasing from 0.16 to 0.02 mg kg-1 over the last five administrations. The 18 remaining bitches were given dexamethasone twice a day for 9.5 days, at a dosage of 0.2 mg kg-1 for 7 days and then decreasing from 0.16 to 0.02 mg kg-1 over the last five administrations. Pregnancy was terminated without complication in 75 of the 80 bitches, and uterine contents were absorbed or aborted or both. Pregnancy was not terminated in five bitches treated for 7.5 days beginning at day 30-35 of gestation. In four of them parturition occurred at the normal time; in one, pregnancy was terminated by a second treatment. Pregnancy termination occurred at 7-15 days after the start of treatment. During ultrasonographic imaging of 13 bitches treated with dexamethasone, fetal deaths occurred between 5 and 13 days after the start of treatment. The side effects reported by the owners included polydipsia and polyuria, which were observed in all of the bitches and which disappeared when the treatment was discontinued. Some bitches also experienced vaginal discharge, restlessness, anorexia or emesis. At the first or second cycle after treatment, 20 bitches were mated and had normal pregnancies and normal litters. The results suggest that oral treatment with dexamethasone can be used to terminate pregnancy in bitches, but that in some cases the withdrawal of treatment after 8 days can result in retention of live pups and require a further treatment or the use of another abortifacient. PMID- 9404291 TI - Prevention of pregnancy in bitches following unwanted mating: a clinical trial using low dose oestradiol benzoate. AB - A field investigation was undertaken in mated bitches to assess the efficacy and safety of a low dose of oestradiol benzoate (0.01 mg kg-1), given by subcutaneous or intramuscular injection, on days 3, 5 and sometimes day 7 after mating. Twenty five veterinary practices in the UK collected data from 358 cases in total. Bitches were treated with oestradiol and then examined at about 65 days after mating, at which time owners were also questioned about any possible adverse signs seen since treatment. Overall 16 bitches (4.5%) became pregnant, although the apparent duration of gestation in some cases suggested that a later mating may have taken place, thus giving a pregnancy rate of only 3.4%. As regards possible side effects of the treatment, 4.5% experienced signs of false pregnancy, 7.8% had evidence of vaginal discharge and 7.3% developed pyometra at some stage during the ten weeks after treatment. None of the bitches showed any clinical signs of bone marrow suppression. The efficacy and safety of this low dose regimen suggest that it is suitable for widespread use, under veterinary supervision. However, it is important for bitch owners to be informed that efficacy can never be guaranteed in any particular case and thus a follow up pregnancy examination may sometimes be advisable. PMID- 9404292 TI - Clinical protocol for pregnancy termination in bitches using prostaglandin F2 alpha. AB - Sixty-seven pregnant bitches were given atropine sulphate (0.025 mg kg-1), prifinium bromide (0.1 ml kg-1) and metopimazine (0.5 mg kg-1) and 15 min later 2.5 micrograms cloprostenol kg-1 s.c., three times at 48 h intervals (day 1, day 3, day 5). After one treatment, 53 of the 67 bitches had aborted, and after a second treatment, 62 of the 67 bitches had aborted. In 18 bitches, progesteronemia kinetics were followed-up: the first injection of cloprostenol resulted in a significant (P < 0.01) fall in progesteronemia. In 12 of the 18 bitches that had aborted following the first protocol, this rapid fall in progesterone was noteworthy as it decreased progesterone concentration on average from 17.07 +/- 8.20 ng ml-1 on day 1 to 1.31 +/- 0.34 ng ml-1 on day 3. The premedication administered 15 min before the injection of prostaglandins, prevented the appearance of side effects in 39 of the 67 bitches (58.2%). PMID- 9404293 TI - Effects of low doses of prostaglandin F2 alpha during the early luteal phase before and after implantation in beagle bitches. AB - Three groups of five beagle bitches were treated three times a day with natural prostaglandin F2 alpha (PGF2 alpha) at a dosage of either 20 micrograms kg-1 bodyweight (days 5-8 of metoestrus), 50 micrograms kg-1 bodyweight (days 5-11 of metoestrus) or 20 micrograms kg-1 bodyweight after detection of pregnancy (days 20-21 after ovulation) for 7 days. A dose of 20 micrograms PGF2 alpha kg-1 bodyweight administered during the early luteal stage could not induce a reliable decrease of progesterone concentrations, while injections of 50 micrograms PGF2 alpha kg-1 bodyweight beginning before implantation resulted in arrest of luteal progesterone production and prevention of nidation in all five bitches. The application of 20 micrograms PGF2 alpha kg-1 bodyweight shortly after implantation induced functional arrest of corpora lutea and led to embryonic or fetal resorption in all cases. In general, the luteolytic effect of low PGF2 alpha doses was insufficient because of the recovery of the corpora lutea seen in nearly all bitches and the prolonged process of embryonic or fetal resorption that increase the risk of uterine disease. PMID- 9404294 TI - Termination of pregnancy in cats using a combination of cabergoline, a new dopamine agonist, and a synthetic PGF2 alpha, cloprostenol. AB - The objective of this study was to increase efficacy of treatment with the dopamine agonist, cabergoline, for inducing abortion in cats by combining it with a synthetic PGF2 alpha analogue, cloprostenol. Side effects of cloprostenol were avoided by using low doses. An oral formulation of cabergoline was chosen to facilitate administration. Cabergoline was given daily (5 micrograms kg-1 day-1) while s.c. injections of cloprostenol were administered every 2 days (5 micrograms kg-1) (2 days)-1). Plasma concentrations of progesterone, side effects and pregnancy outcome were compared with those of five untreated pregnant queens. The treatment was administered from day 30 after first mating in five queens confirmed as pregnant and lasted a mean of 11 +/- 1 days. All treated animals aborted in 9 +/- 1 days without any side effect, except a mild haemorrhagic vulvar discharge. Subsequent fertility of the queens was not compromised. Abortion was mediated by an abrupt and constant decrease in plasma concentrations of progesterone. Progesterone concentrations were lower than 1 ng ml-1 from day 38 onwards in all treated animals and remained at this value thereafter, except for one queen which showed a further increase in plasma progesterone concentration from day 44 without any clinical consequence. In conclusion, a combination of daily oral administration of cabergoline and cloprostenol injections every 2 days appears to be a reliable, safe and practical method for terminating pregnancy in cats at day 30 of pregnancy, when a diagnosis of pregnancy by palpation or ultrasonography can easily be made. PMID- 9404295 TI - Urinary LH, plasma LH and progesterone and their clinical correlates in the periovulatory period of domestic bitches. AB - Ten bitches were examined daily from pro-oestrus until metoestrus to correlate endocrine, vaginoscopic and cytological assessments of stage of the oestrous cycle. It was considered that oocytes were fertile by 4 days after the LH peak assuming a 2 day lag before ovulation and a further 2 days for oocyte maturation. Eight bitches showed a plasma LH peak and five bitches showed a peak in urinary LH, which corresponded to the plasma LH peak. Plasma concentration of progesterone increased in all bitches. Initially, this coincided with the preovulatory LH peak and reached values of 7.7 +/- 0.6 ng ml-1 4 days later. The peak in number of a nuclear vaginal epithelial cells and the onset and peak of vaginal mucosal shrinkage with angulation occurred on average 2.4 +/- 1.5, 2.1 +/ 2.4, and 6.1 +/- 1.1 days after the plasma LH peak, respectively, giving sufficient precision to be of practical value in estimating the fertile period. However, the onset of vulval softening, changes in rectal temperature and colour of the vulval discharge varied considerably. Nine of the bitches were mated at the time of highest percentage of a nuclear cells, coinciding with the onset of the period of vaginal mucosal shrinkage with angulation. Eight became pregnant. Selected clinical parameters are therefore useful in predicting the optimal mating time of bitches, as is detection of plasma progesterone concentration higher than about 8 ng ml-1, but the latter appears to be the most reliable method. Detection of the urinary LH peak was more difficult and as yet offers no practical advantages except from the point of view of easy sample collection. PMID- 9404296 TI - Use of an immunoenzymatic assay to detect the luteinizing hormone peak in bitches. AB - The success of artificial insemination with fresh chilled or frozen semen depends on the time of insemination. Determination of oestrous behaviour, use of vaginal smears or measurement of the vaginal resistivity are unreliable techniques and interpretation of the dosages of progesterone may be critical as it may vary from one laboratory to another. The plasma concentration of LH displays a peak of secretion that can be a good reference to date the events of the ovarian cycle. The plasma concentration of LH was measured in ten bitches by a new sandwich immunoenzymatic assay (Reprokit, Sanofi) and results were compared with the vaginal smears and plasma concentrations of progesterone. The LH peak was easily detected and lasted 3.3 days with maximum values ranging from 10 to 22 ng ml-1. At the time of the peak, the vaginal smears displayed features of pro-oestrus and the plasma concentration of progesterone was 2.9 ng ml-1. This immunoenzymatic assay, already used for many species, is an effective tool for determining the time of the LH peak. Added to the study of the progesterone concentration, it may help to determine the optimal time for artificial inseminations, particularly when frozen semen is used. PMID- 9404297 TI - New techniques using transcervical uterine cannulation for the diagnosis of uterine disorders in bitches. AB - A technique for collecting uterine samples from bitches without the need for surgery was developed. This technique involved visualizing the cervix with a rigid endoscope and passing a catheter through the cervix into the uterus. Samples for microbiology and cytology were obtained by the infusion and aspiration of sterile normal saline. This technique allowed uterine microbiology and cytology of the normal bitch throughout the reproductive cycle. Microorganisms were frequently recovered from the uterus during pro-oestrus and oestrus, but rarely at other stages of the reproductive cycle. The uterine microflora often reflected the vaginal microflora during pro-oestrus and oestrus. The cells found in uterine cytology samples from normal bitches included endometrial epithelial cells, leukocytes, erythrocytes, cervical cells, spermatozoa and bacteria. The types, proportions, morphology and numbers of cells varied throughout the reproductive cycle. The endoscope could be passed into the uterus and the endometrium examined from parturition until day 23 post partum. These procedures and contrast hysterography were used to investigate the reproductive tract of bitches. The above techniques have facilitated the diagnosis of postpartum metritis (n = 3), pyometra (n = 2), endometritis (n = 1), abortion (n = 1), retained placenta (n = 1), postpartum uterine rupture (n = 1), endometrial subinvolution (n = 1) and misalliance (n = 1) in 21 bitches investigated. PMID- 9404298 TI - Plasma concentrations of prolactin in overtly pseudopregnant Afghan hounds and the effect of metergoline. AB - The effect of metergoline, a 5-hydroxytryptamine (serotonin) antagonist, on the plasma concentrations of prolactin in overtly pseudopregnant Afghan hounds and on the clinical symptoms of overt pseudopregnancy were studied. Plasma concentrations of prolactin and progesterone were determined in six Afghan hounds with signs of overt pseudopregnancy for 2-3 weeks and in three Afghan hounds that were not pseudopregnant at the time of blood sampling. In the overtly pseudopregnant bitches the plasma concentrations of prolactin before treatment (35.5 +/- 8.5 micrograms l-1) were significantly higher than the plasma concentrations of prolactin of the three bitches that were not pseudopregnant (6.3 +/- 0.5 micrograms l-1); the latter values were similar to those of non pseudopregnant beagle bitches during the total luteal phase. The six pseudopregnant Afghan hounds were treated for 10 days with the antiserotoninergic drug metergoline. At 2 h after the onset of treatment with metergoline, the mean plasma concentration of prolactin had decreased to 10.8 +/- 2.9 micrograms l-1. The plasma concentrations of prolactin continued to decline to 5.4 +/- 1.0 micrograms l-1 at 4 h and to 1.0 +/- 0.1 microgram l-1 during treatment days 3 10. Signs of pseudopregnancy, such as swelling of the mammary glands and digging, decreased during the treatment period. The treatment was associated with mild behavioural side effects such as whimpering and aggressiveness. These side effects are probably not related to suppression of prolactin but are due to a direct effect on serotoninergic pathways in the brain. It is concluded that high plasma concentrations of prolactin are associated with the development and maintenance of pseudopregnancy. The serotonin antagonist metergoline strongly suppresses plasma concentrations of prolactin in pseudopregnant dogs and decreases the clinical signs of pseudopregnancy. PMID- 9404299 TI - Transrectal ultrasonographic examination of the female urogenital tract in nonpregnant and pregnant captive bears (Ursidae). AB - The development of reliable methods for monitoring the reproductive cycle of bears is needed to optimise breeding management in captivity. The aim of the present study was to develop non-invasive procedures for obtaining basic data on the reproductive status of nonpregnant and pregnant bears. Eight female captive bears (five Ursus arctos arctos, two Tremarctos ornatus and one Melursus ursinus) were examined using transrectal adaptor ultrasonographic imaging. Plasma concentrations of progesterone and gestagens in faeces were determined simultaneously. The ultrasonographic appearance of the female urogenital tract was assessed during the nonbreeding season and verified in four adult bears post mortem. Pregnant bears were examined twice, as the reproductive physiology of bears is characterized by delayed implantation and pseudopregnancy. During embryonic diapause there was no difference between pseudopregnant and pregnant bears both by ultrasonographic imaging and endocrinological data. After implantation (early December) the pregnancies were visualised by ultrasonographic imaging, whereas plasma concentrations of progesterone and gestagens in faeces had not yet increased. In conclusion, both transrectal ultrasonographic imaging and gestagen monitoring in faeces are efficient methods for obtaining important data on reproduction in bears. PMID- 9404300 TI - Feline colostrum--friend or foe: maternal antibodies in queens and kittens. AB - The transfer of immunoglobulins (Ig) by colostrum from the queen to the neonatal kitten not only provides protection from infection, but may also cause serious illness. Neonatal isoerythrolysis may occur when kittens of blood type A or AB receive colostral anti-A alloantibodies from a type B queen. In contrast to other species, Ig concentrations in milk and colostrum did not differ markedly. Gastrointestinal absorption of IgG was limited to the first day of life. The half lifes of maternally derived IgG and IgA in kittens were shorter than in puppies. In conclusion, milk from another queen may be given as a replacement for colostrum to neonatal kittens. Kittens at risk of neonatal isoerythrolysis must be removed from their type B queen during the first day of life and may safely receive milk or colostrum from a type A queen. PMID- 9404301 TI - Hormonal state and effects of the use of an antiprogestin in bitches with pyometra. AB - The pathogenic significance of progesterone in pyometra in bitches was investigated by evaluating the stages of the oestrous cycles of 369 bitches with pyometra. Concentrations of progesterone and oestradiol were determined in 100 bitches with pyometra before ovariohysterectomy. Six bitches with pyometra with progesterone > 5 ng ml-1 and oestradiol < 25 pg ml-1 were treated with the antigestagen RU 46534 (6 mg kg-1, s.c.) twice on day 1 and once on days 2, 3 and 4; ovariohysterectomy was performed on day 6. Six control bitches that met the same criteria were ovariohysterectomized and uterine morphology was compared with that of the treated bitches. The effects of treatment were monitored by hormone, ultrasonographic imaging and histomorphological studies. The results confirm that pyometra may occur at any stage of the reproductive cycle and that most animals are in the phase of dioestrus. Uterine mass (g kg-1 bodymass) with versus without content was not different in treated bitches and was significantly lower (P < 0.01) than in the control bitches. In all six dogs general clinical conditions improved during the observation period; 23.5 +/- 11.1 h after the start of antiprogestin treatment a profuse, purulent vulval discharge was observed. The effusion was nearly complete at day 6. The maximal detectable uterine lumen varied between 23 and 39 mm in diameter, it was no longer detectable on day 6. Oestradiol concentrations remained constant while progesterone decreased with distinct individual variations. Morphometrical examination of the endometrium revealed no differences between groups. The data suggest that antigestagen treatment is a promising approach for treatment of pyometra in bitches. PMID- 9404302 TI - Treatment of pyometra (cystic endometrial hyperplasia) in bitches with an antiprogestin. AB - On the basis of a previous study showing the effectiveness of an antiprogestin treatment on the involution of the pyometric uterus and general health of bitches, in the present open clinical study seven bitches with pyometra and progesterone concentrations > or = 2 ng ml-1 were treated with the antiprogestin RU 46534. The dose selected was 5 or 6 mg kg-1 body mass s.c., on the first day of treatment and 3 mg kg-1 body mass on days 2, 3, 4, 8, 12 and 16. Antibiotics were administered until day 16. A vulval discharge was observed within 12-24 h. In one bitch the dose of the antiprogestin had to be increased after day 4. In six bitches the uterine lumen became ultrasonographically undetectable between days 8 to 12; in one bitch some luminal material could still be detected on day 28. The number of blood leucocytes tended to increase after the onset of treatment but had returned to the upper normal range by day 16. In all dogs general condition and feed consumption improved rapidly and were normal within 8 days. No side effects were noted. Two of the dogs were mated subsequently and produced two healthy litters. These observations confirm those of a preceding study and show that the treatment of pyometra with an antiprogestin may lead to a clinical recovery. These results justify further studies to enable the optimum treatment regimen to be established. PMID- 9404303 TI - Progestin-induced hypersecretion of growth hormone: an introductory review. AB - In the 1970s acromegalic features were reported in some dogs used in long-term toxicity studies of progestins. In 1980 confirmation that progestagen administration can lead to increased circulating growth hormone (GH) concentrations was obtained. This phenomenon appeared not to be confined to exogenous progestins, for an excess of GH was also found in bitches during the luteal phase of the oestrous cycle. In bitches with a progestin-induced excess of GH, GH secretion could neither be inhibited nor stimulated by well-known regulatory neurohormones, indicating autonomous secretion. Because it could not be attributed to a neoplasm and was reversible, an extra-pituitary site of GH production was investigated. The progestin-induced GH was found to originate from the mammary gland. This phenomenon seems to play a role in the mammary development that occurs during the luteal phase of the oestrous cycle. The increase in cell proliferative activity may also be responsible for the susceptibility of the mammary gland to neoplastic transformation. The discovery of mammary GH in the dog has recently become of wider importance now that expression of the GH gene has also been demonstrated in other species, namely, humans and cats. PMID- 9404304 TI - The role of progestins, insulin-like growth factor (IGF) and IGF-binding proteins in the normal and neoplastic mammary gland of the bitch: a review. AB - The growth hormone (GH) and insulin-like growth factor (IGF) system is an important regulatory system of mammalian epithelial cell proliferation and differentiation. The biological effects of the IGFs are modulated by six different binding proteins (IGFBPs). Progestins play an important role in the regulation of the dynamics of mammary gland development and involution through the modulation of these growth regulating factors. In dogs and cats, progestins stimulate the local production of GH in the mammary gland. In dogs, this results in high plasma concentrations of GH and a concomitant increase in plasma IGF-I and IGFBP-3 concentrations. The administration of progestins also induces high plasma concentrations of IGF-II, even before GH concentrations start to increase. In the mammary gland of the normal bitch, IGFBP-5 and IGFBP-2 are the main IGFBPs expressed. Progestin administration results in a decrease of mRNA encoding IGFBP 5, but does not alter the concentration of mRNA encoding IGFBP-2. This local mammary system of GH, IGFs and IGFBPs plays an important role in the regulation of mammogenesis, lactation and involution. However, the presence of a high proliferative environment may also enhance the risk of malignant transformation and promotion of tumour growth with an associated inhibition of programmed cell death. PMID- 9404305 TI - Effects of progestin administration on the hypothalamic-pituitary-adrenal axis and glucose homeostasis in dogs. AB - The effects of medroxyprogesterone acetate (MPA) and proligestone (PROL) on the hypothalamic-pituitary-adrenocortical axis and glucose homeostasis were studied in two groups of eight ovariohysterectomized beagle bitches. In addition, the binding characteristics of MPA and PROL for the progesterone and glucocorticoid receptor were investigated. The administration of both progestins resulted in suppression of the hypothalamic-pituitary-adrenal axis. Whereas basal plasma concentrations of adrenocorticotrophic hormone (ACTH) were only moderately affected, the basal plasma concentrations of cortisol and the cortisol:creatinine ratio in urine were significantly decreased after the first administration of both progestins. In the group given MPA the increase of ACTH after stimulation with corticotrophin-releasing hormone (CRH) remained normal but it was suppressed in the group treated with PROL. In both treatment groups the increase of cortisol after stimulation with CRH was lower. After cessation of progestin administration both basal and stimulated plasma ACTH concentrations returned to pretreatment concentrations within a few weeks. In contrast, it took 6 month to restore the basal plasma cortisol concentrations and cortisol:creatinine ratios in urine. Paradoxically, the stimulated cortisol concentrations returned to normal shortly after the cessation. Histological examinations revealed a severe atrophy of the zona fasciculata and reticularis of the adrenal gland in all treated dogs and a steroid-induced hepatopathy in 50% of them. During the first half of the progestin treatment, glucose homeostasis was maintained by increased plasma concentrations of insulin in both groups. After prolonged treatment the response to a glucose load became impaired. None of these parameters improved during the 6 month recovery period. Histological changes in the pancreas, characteristics of diabetes mellitus, were found in two dogs of each group. Most probably, the glucocorticoid action of the progestins is not the sole explanation for the insulin resistance since progestin treatment resulted in a concomitant increase in plasma concentrations of growth hormone which has diabetogenic properties. Experiments in vitro confirmed the strong glucocorticoid component of MPA and PROL. The inhibition constants (Ki) of PROL for both the progesterone receptor (PR) and the glucocorticoid receptor (GR) were approximately then times higher than those of MPA. Nonetheless, the ratios of the Ki for the GR and PR indicated that the specificity of MPA and PROL was only slightly different but considerably smaller than that of progesterone. It is long-term treatment with high doses of these progestins may result in a iatrogenic Cushing's syndrome and diabetes mellitus. PMID- 9404306 TI - Lack of association of progestin-induced cystic endometrial hyperplasia with GH gene expression in the canine uterus. AB - Growth hormone (GH) is produced in progestin-induced hyperplastic ductular mammary epithelia in dogs. Progestins also induce the development of cystic endometrial hyperplasia (CEH) in this species. The study reported here investigated whether GH gene expression could also be demonstrated in progestin induced hyperplastic epithelium in the canine uterus. Eight beagle bitches were treated with 10 mg medroxyprogesterone acetate (MPA) kg-1 body mass s.c. at intervals of 3 weeks, for a total of five times in four dogs (group I) and for a total of 13 times in the other four dogs (group II). Blood samples were collected twice during each 3 week period for measurement of plasma concentrations of GH, insulin-like growth factor I (IGF-I) and IGF-II. At the end of the series of injections uterine tissue was obtained by ovario-hysterectomy. Histological examination confirmed that CEH was present in all uteri after MPA treatment; the changes in the dogs of group I were less marked than those in group II. Immunohistochemical examination of the uterine tissues showed that immunoreactive(i) GH was present in a number of uteri with CEH. iGH was usually located in the cytoplasm of glandular epithelial cells. However, reverse transcriptase PCR using GH-specific primers failed to demonstrate mRNA encoding GH in the uterine tissue of all dogs. It is concluded that local production of GH is not involved in progestin-induced hyperplasia of uterine epithelial cells in dogs. PMID- 9404307 TI - Growth hormone concentrations in mammary secretions and plasma of the periparturient bitch and in plasma of the neonate. AB - The presence of growth hormone (GH) in mammary secretions of bitches was investigated in relation to plasma GH concentrations at about the time of parturition and during the first weeks of lactation. Plasma GH concentrations in neonates were measured during the first weeks of lactation, to determine whether GH in maternal milk contributes to plasma concentrations of GH in the neonate. Gastrointestinal uptake of GH was studied by measurement of plasma bovine GH (bGH) concentrations after intragastric administration of bGH. High concentrations of GH were found in the mammary secretions of the bitches, particularly before parturition and in colostrum, exceeding maternal plasma concentration up to 100-1000 times. GH concentrations in milk were not not significantly correlated with GH concentrations in plasma of bitches or neonates. Bovine GH could not be detected in neonatal plasma for 4 h after intragastric administration of bGH. The presence of high concentrations of canine GH (cGH) in ante-partum and colostral mammary secretions is consistent with the progesterone induced mammary biosynthesis of GH. GH in milk is probably not absorbed from the gastrointestinal tract into the blood circulation of the newborn in its intact form. PMID- 9404308 TI - Geriatricians: past, present and future. PMID- 9404309 TI - The politics of disability: some implications for geriatricians. PMID- 9404310 TI - Can a medical and social assessment be combined? PMID- 9404311 TI - Lewy body dementia. PMID- 9404312 TI - Alcohol problems in the older person. PMID- 9404313 TI - Frailty: help or hindrance? PMID- 9404314 TI - Personality, health and ageing. PMID- 9404315 TI - Sarin poisoning of a rescue team in the Matsumoto sarin incident in Japan. AB - OBJECTIVES: A nerve agent sarin (isopropyl methyl phosphonofluoridate) was released in Matsumoto city, Japan, on 27 June 1994. About 600 people were affected by the sarin, including seven who died. Fifty two rescuers engaged in helping the victims and 18 were affected. The aim was to investigate how the rescuers were affected by sarin. METHODS: Health examinations and a questionnaire survey were conducted with all rescuers. RESULTS: A rescuer who was one of the first engaged and who worked for about five hours in areas contaminated with sarin was admitted to hospital after poisoning; the others did not consult doctors although they showed slight muscarinic symptoms. The later the rescuers started their work, the less likely they were to experience symptoms of sarin exposure, and no one starting work 270 minutes after the original release of sarin was affected. The symptoms of exposure included ocular pain, darkness of visual field, nausea, vomiting, headache, rhinorrhea, narrowing of visual field, sore throat, fatigue, and dyspnoea, which were similar to those reported by the citizens who were sarin victims. There were no rescuers who had abnormal physical or neurological signs associated with sarin at the time of the physical examination conducted three weeks after the sarin release. A year after the sarin incident, the symptoms of all the rescuers had resolved. CONCLUSIONS: Rescuers should protect themselves with appropriate clothing, gloves, and a mask to prevent a secondary disaster for at least 24 hours after a similar accident. PMID- 9404316 TI - Mortality of workers exposed to dieldrin and aldrin: a retrospective cohort study. AB - OBJECTIVE: To investigate the occurrence of long term health effects in humans exposed to aldrin and dieldrin, with an update of an earlier retrospective cohort mortality study. METHODS: A group of 570 workers employed between 1 January 1954 and 1 January 1970 either in a production or formulation plant were followed up for mortality until 1 January 1993. There were extensive industrial hygiene data available and biological monitoring data of aldrin and dieldrin for most of the workers. From these data individual estimates were made of the total intake of dieldrin. A total number of 2539.37 person-years at risk was added to the original study. RESULTS: 118 deaths were observed compared with 156 expected. No increase in mortality from liver cancer was found. However, there was an excess in mortality from rectal cancer. This excess was inversely related to the dose gradient. An analysis by job title did not show any excess cancer in any particular job. CONCLUSION: The study does not support a carcinogenic effect of dieldrin and aldrin in humans. PMID- 9404317 TI - Mortality of Dutch coal miners in relation to pneumoconiosis, chronic obstructive pulmonary disease, and lung function. AB - OBJECTIVES: To analyse the mortality patterns of former Dutch coal miners, focusing on coal workers' pneumoconiosis (CWP) and chronic obstructive pulmonary diseases (COPD) in relation to pre-existing impairment of lung function. METHODS: 3790 selected miners, medically examined between 1952 and 1963, were followed up to the end of 1991 with the municipal population registries and the causes of death from the death certificates were ascertained and converted to the codes from the ninth revision of the international classification of diseases (ICD-9). Mortality comparisons were made with the male population in The Netherlands, resulting in standardised mortality ratios (SMRs). 3367 miners had radiological manifestation of CWP at medical examinations. RESULTS: 80% of the miners died during the follow up period. Excess mortalities from CWP (SMR 4523) and COPD (SMR 179) were found. Coal miners without CWP also showed an increased mortality from COPD (SMR 2913). A diminished lung function (forced expiratory volume in one second (FEV1), or FEV1/FVC (forced vital capacity) ratio) at medical examination resulted in a significantly increased SMR for COPD (322 and 212 respectively) whereas normal lung function yielded expected mortalities from COPD. A positive correlation also emerged between diminished lung function and the SMR due to CWP. The body mass index (BMI) at the moment of medical examination was correlated with the risk of dying of COPD and CWP: a decreasing BMI resulting in an increased SMR. CONCLUSIONS: Not only infectious diseases and CWP but also COPD is an important cause of occupational mortality in miners with extensive exposure to coal mine dust. No obvious connection between pre-existing CWP and the COPD mortality exists. Impaired FEV1 and FEV1/FVC ratios are predictors of an increased risk of COPD death. The BMI seems to indicate the severity of the COPD, resulting in premature death. PMID- 9404318 TI - Efficiency of different grouping schemes for dust exposure in the European carbon black respiratory morbidity study. AB - OBJECTIVES: The aim of this study was to assess the theoretical efficiencies of different grouping strategies and its effect on the exposure-response relation in a study of respiratory morbidity associated with exposure to total inhalable and respirable carbon black dust. METHODS: A large epidemiological study is being undertaken to investigate the respiratory health of employees in the European carbon black manufacturing industry in relation to exposure to carbon black dust. In phase 2 of the study, repeated measurements of total inhalable and respirable dust were taken which enabled estimation of various components of variability in the exposure data (within and between worker variance and within and between group variance). These variance components were used to calculate the contrast in exposure between the groups in various classification schemes and to calculate the theoretical attenuation of the exposure-response relation and the standard error (SE) of the slope. RESULTS: High contrast in exposure was found when workers were classified according to the combination of their factory and job category as well as when these combinations were amalgamated into five exposure groups. Attenuation was minimal with most grouping schemes; only with the individual based strategy was the attenuation large. The SE of the theoretically attenuated exposure-response slope was smallest for the strategy based on individual people followed by the classification scheme based on factory and job category. CONCLUSIONS: It was concluded that, although some assumptions for the calculations of the attenuation of the exposure-response slope were not met, the most appropriate classification scheme of the worker seems to be by the combination of factory and job category. PMID- 9404319 TI - Cancer mortality among electric utility workers exposed to polychlorinated biphenyls. AB - OBJECTIVES: To assess whether excess mortality from cancer, malignant melanoma of the skin, and cancers of the brain and liver in particular, is associated with long term occupational exposure to polychlorinated biphenyls (PCBs). METHODS: An epidemiological study of mortality was conducted among 138,905 men employed for at least six months between 1950 and 1986 at five electrical power companies in the United States. Exposures were assessed by panels composed of workers, hygienists, and managers at each company, who considered tasks performed by workers in 28 job categories and estimated weekly exposures in hours for each job. Poisson regression was used to examine mortality in relation to exposure to electrical insulating fluids containing PCBs, controlling for demographic and occupational factors. RESULTS: Neither all cause nor total cancer mortality was related to cumulative exposure to PCB insulating fluids. Mortality from malignant melanoma increased with exposure; rate ratios (RRs) relative to unexposed men for melanoma were 1.23 (95% confidence interval (95% CI) 0.56 to 2.52), 1.71 (0.68 to 4.28) and 1.93 (0.52 to 7.14) for men with < 2000, > 2000-10,000, and > 10,000 hours of cumulative exposure to PCB insulating fluids, respectively, without consideration of latency. Lagging exposure by 20 years yielded RRs of 1.29 (0.76 to 2.18), 2.56 (1.09 to 5.97), and 4.81 (1.49 to 15.50) for the same exposure levels. Mortality from brain cancer was modestly increased among men with < 2000 hours (RR 1.61, 95% CI 0.86 to 3.01) and > 2000-10,000 hours exposure (RR 1.79, 95% CI 0.81 to 3.95), but there were no deaths from brain cancer among the most highly exposed men. A lag of five years yielded slightly increased RRs. Mortality from liver cancer was not associated with exposure to PCB insulating fluids. CONCLUSIONS: This study was larger and provided more detailed information on exposure than past investigations of workers exposed to PCBs. The results suggest that PCBs cause cancer, with malignant melanoma being of particular concern in this industry. PMID- 9404320 TI - Risk of cancer among paper recycling workers. AB - OBJECTIVES: Studies in traditional paper mills have indicated an excess cancer risk, and mutagenic compounds have been identified in the industry. No studies have reported on risk of cancer in paper recycling. Therefore the cancer incidence in Danish paper recycling mills was investigated. METHODS: 5377 employees in five paper recycling plants were included in a historical cohort study. The workers had been employed in paper recycling in 1965-90, and the cohort was followed up until 31 December 1993. The expected number of cancer cases was calculated from national rates. RESULTS: There was significantly more pharyngeal cancer among male workers (seven observed (standardised incidence ratio (SIR) 3.33, 95% confidence interval (95% CI) 1.34 to 6.87)). There was slightly more lung cancer among male workers in production (39 observed, SIR 1.21, 95% CI 0.86 to 1.65). Risk of Hodgkin's disease was doubled in male production worker (four observed, SIR 1.90, 95% CI 0.51 to 4.85). CONCLUSIONS: The increased risk of pharyngeal cancer found in this study is interesting but may be influenced by confounders such as smoking and alcohol intake. This study also indicates an excess risk of Hodgkin's disease, which is in accordance with some studies in the traditional paper mills. As this is the first report on risk of cancer in paper recycling, further studies are needed. PMID- 9404321 TI - Risk factors for carpal tunnel syndrome in a general population. AB - OBJECTIVE: To determine the individual, physical, and psychosocial risk factors for carpal tunnel syndrome in a general population. METHODS: Population based case-control study in Marshfield epidemiological study area in Wisconsin, USA. Cases were men and women aged 18-69 with newly diagnosed carpal tunnel syndrome (n = 206 (83.1%) of 248 eligible). Controls were a random sample of residents of the study area who had no history of diagnosed carpal tunnel syndrome (n = 211 (81.5%) of 259 eligible). Cases and controls were matched by age. Telephone interviews and reviews of medical records obtained height and weight, medical history, average daily hours of exposure to selected physical and organisational work factors, and self ratings on psychosocial work scales. RESULTS: In the final logistic regression model, five work and three non-work variables were associated with risk of carpal tunnel syndrome, after adjusting for age. For each one unit of increase in body mass index (kg/m2), risk increased 8% (odds ratio (OR) 1.08; 95% confidence interval (95% CI) 1.03 to 1.14). Having a previous musculoskeletal condition was positively associated with carpal tunnel syndrome (OR 2.54; 95% CI 1.03 to 6.23). People reporting the least influence at work had 2.86 times the risk (95% CI, 1.10 to 7.14) than those with the most influence at work. CONCLUSIONS: Carpal tunnel syndrome is a work related disease, although some important measures of occupational exposure, including keyboard use, were not risk factors in this general population study. The mechanism whereby a weight gain of about six pounds increases the risk of disease 8% requires explanation. PMID- 9404322 TI - Work environment and low back pain: the influence of occupational activities. AB - OBJECTIVES: To find associations between the prevalence of low back pain and occupational activities. METHODS: Interviews of a random sample of 5185 19-59 year old Danish employees analysed by logistic regression. RESULTS: Increased risks of low back pain were found for "vibration affecting the whole body" (odds ratio (OR) = 1.28), "physically hard work" (OR = 1.28), "frequently twisting or bending" (OR = 1.71), "standing up" (OR = 1.20), and "concentration demands" (OR = 1.28). In the analysis of dose-response relations between low back pain and the risk factors, the one year period prevalence increased with increasing exposure time during a working day to each of the risk factors. The prevalence proportion ratio for those reporting to be exposed for most of the working time were 1.30 for vibrations affecting the whole body, 1.54 for physically hard work, 1.48 for frequently twisting or bending, 1.29 for standing up, and 1.13 for concentration demands. These associations seemed to be stronger in the subset of subjects who worked for 37 hours or more per week. The population attributable fractions were 15.1% for frequently twisting or bending, 15.0% for standing up, 7.6% for concentration demands, and 4.4% for physically hard work. CONCLUSION: Vibrations affecting the whole body, physically hard work, frequently twisting or bending, standing up, and concentration demands proved to be risk factors for the occurrence of low back pain, even after controlling for age, sex, educational level, and duration of employment in a specific occupation. PMID- 9404323 TI - Neurological signs in relation to cancer in patients with asbestosis. AB - OBJECTIVE: To chart the subtle neurological abnormalities in patients with asbestosis relative to possible development of cancer. METHODS: In 1979-81 a standardised neurological examination was made of 115 patients with asbestosis who carried a high risk of occupational cancer and their cancer morbidity was analysed 15 years later. RESULTS: Slight disturbances of unknown aetiology were found in the central nervous system (CNS) of 33 and in the peripheral nervous system (PNS) of 41 patients. Of these 17 had disturbances of both the CNS and PNS. This cohort was followed up to the end of 1994. During this time 47 of the patients developed cancer. Statistical analyses showed that disturbances of the CNS such as psycho-organic syndrome, cerebellar dysfunction, and motor disturbances of unknown origin were significantly associated with cancer, whereas no such association was found for peripheral neuropathy. Interaction between the radiological progression of asbestosis and disturbances of the CNS was an even stronger predictor of cancer. CONCLUSIONS: It seems that slight disturbances of the CNS are predictors of development of cancer. Whether or not these disturbances are manifestations of involvement of a paraneoplastic nervous system or some factor associated with progression of asbestosis remains open. PMID- 9404324 TI - Concentration of cadmium in rice and urinary indicators of renal dysfunction. AB - OBJECTIVES: (1) To examine the relation between concentrations of cadmium (Cd) in rice and urinary concentrations of indicators of renal dysfunction and the prevalence of abnormalities in urine in areas polluted by Cd. (2) To establish the maximum allowable concentration of Cd in rice from these findings. METHODS: The target population consisted of 1703 inhabitants (832 men and 871 women) aged over 50 years who consumed home grown rice and had lived in the same hamlet in areas polluted by Cd in the Kakehashi River basin in Ishikawa Prefecture, Japan for at least 30 years. The correlation coefficients between concentrations of Cd in rice and several urinary substances, the prevalence of abnormalities in urine and sex in hamlets polluted by Cd were calculated. Finally, regression analysis was performed for significant indicators to calculate the maximum allowable concentration of Cd in rice based on values in a control group. CONCLUSIONS: Significant correlations between concentration of Cd in rice and concentrations of urinary beta 2-microglobulin, metallothionein, glucose, and aminonitrogen were established. Similarly, there were significant correlations between concentration of Cd in rice and the prevalence of beta 2-microglobulinuria, metallothioneinuria, glucosuria, proteinuria, proteinuria with glucosuria, and aminonitrogenuria. The highest maximum allowable concentration of Cd in rice calculated for these indicators was 0.34 ppm/l and 0.29 ppm/g creatinine. Both values are lower than 0.4 ppm, the tentative limit prescribed by the Japanese government. PMID- 9404325 TI - Outcome determinants for isocyanate induced occupational asthma among compensation claimants. AB - OBJECTIVES: To compare the outcome of occupational asthma (OA) induced by isocyanates in Ontario (where a surveillance programme for exposed workers has been in place for over 15 years), with the outcome of OA induced by other work agents. METHODS: Compensated OA claims during the period 1984-88 in Ontario were retrospectively reviewed in a standardised way. RESULTS: 136/235 compensated claims were attributed to isocyanates. Compared with other causes of OA, those attributed to isocyanates had a shorter latent period before onset (5.9 v 7.9 years, P < 0.05), shorter duration of symptoms before diagnosis (2.0 v 3.0 years, P < 0.05), and less associated atopy (43% v 58%, P < 0.05). Outcome at a mean of 1.9 years after initial assessment was significantly better in those with OA induced by isocyanates; 73% cleared or improved v 56% with other causes of OA (P < 0.05). Ten subjects with OA induced by isocyanates stayed at the same work; none cleared and four had worsened at follow up. A better outcome in OA induced by isocyanates was associated with early diagnosis (P < 0.05), and early removal from isocyanates after the onset of asthma. CONCLUSIONS: The outcome in the group with OA induced by isocyanates is similar to previous follow up studies. However, it is better than the outcome in our comparison group with OA due to other causes, perhaps because of earlier diagnosis in the group with OA induced by isocyanates. This may be attributable to the medical surveillance of workers exposed to isocyanates in Ontario, either directly from the surveillance assessments, or indirectly by increasing awareness of the condition. PMID- 9404326 TI - Occupational asthma due to porcine pancreatic amylase. AB - A case of occupational asthma in a 41 year old histopathology laboratory technician attributable to a powder preparation of the porcine pancreatic enzyme amylase is reported. The diagnosis was confirmed by a double blind, placebo controlled, inhalation challenge study which showed immediate and late asthmatic reactions associated with a significant increase in airway responsiveness to methacholine. PMID- 9404327 TI - Predictive value of nerve conduction studies. PMID- 9404328 TI - Prolonged exposure to an epoxy resin leading to interstitial nephritis. PMID- 9404329 TI - Complementary medicine: finding a balance. PMID- 9404330 TI - Do common sources of dietary protein increase calcium needs? PMID- 9404331 TI - New ADA campaign seeks more cosponsors for Medicare Medical Nutrition Therapy Act; update on child/elderly bills. PMID- 9404332 TI - Complementary and alternative medicine: friend, foe, or OWA? PMID- 9404334 TI - A Food Variety Index for Toddlers (VIT): development and application. AB - OBJECTIVE: To develop a variety index based on the Food Guide Pyramid that is specific to toddlers and is indicative of dietary adequacy. DESIGN: Subjects' mothers were assigned randomly to two in-home interviews with a registered dietitian at four possible collection periods: 24, 28, 32, or 36 months. Three days of dietary information were collected at each period. A Variety Index for Toddlers (VIT) was developed to assess variety within and among food groups based on the number of servings from the food groups in the Food Guide Pyramid. SUBJECTS: White children aged 24 to 36 months (n = 124) and their mothers who were participants in an ongoing longitudinal study. STATISTICAL ANALYSES: Descriptive statistical procedures were performed on VIT scores. Mean adequacy ratio (MAR) scores were calculated for all subjects and compared with VIT scores. RESULTS: Bread group scores were consistently the highest of the individual food groups (mean score = 0.94 to 0.96 on the 0.0 to 1.0 scale); the vegetable and meat groups were generally the lowest (mean score = 0.68 to 0.73 and 0.73 to 0.76, respectively). Mean VIT scores (an average of the five food group scores) over the four collection periods ranged from 0.79 +/- 0.14 to 0.81 +/- 0.15; a score of 1.00 represented intake of at least the minimum number of recommended servings from each food group. VIT scores were strongly correlated to the MAR score of nutrient adequacy (r = +.74, P < .01). APPLICATIONS: The VIT can provide a numeric description of dietary variety specific to toddlers. VIT scores can be compared with other characteristics of children, and this index has the potential to be adapted for use with other age groups and populations. PMID- 9404333 TI - Serum and plasma markers of nutritional status in children infected with the human immunodeficiency virus. AB - OBJECTIVE: To determine whether reduced serum or plasma protein and micronutrient levels are common in children infected with the human immunodeficiency virus (HIV) and whether these levels are different in children with growth retardation compared to those with normal growth. SUBJECTS: Children were separated into three groups: (a) HIV-infected with growth retardation (HIV + Gr); (b) HIV infected with normal growth (HIV+); (c) HIV-uninfected with normal growth (HIV-). All children were afebrile and free of acute infection at the time of study. During a 24-hour stay in the Pediatric Clinical Research Unit, blood was drawn for analysis of total protein, albumin, zinc, selenium, and vitamin A levels; growth measurements were obtained; and dietary intake was assessed by 24-hour weighed food intake and 24-hour dietary recall. STATISTICAL ANALYSIS: Mean differences between groups were assessed by analysis of variance, and differences in the frequency of nutrient deficiency were determined by chi 2 analysis. RESULTS: Thirty-eight children between 2 and 11 years of age were studied: 10 HIV + Gr, 18 HIV+, and 10 HIV-. No statistically significantly differences were noted in mean levels of albumin, prealbumin, zinc, and selenium. Mean serum level of vitamin A was significantly higher in the HIV + Gr group than in the other two groups. There were no significant differences between groups in the frequency of deficiency for any nutrient studied. Mean energy and nutrient intake was similar among groups. APPLICATIONS/CONCLUSIONS: Abnormal serum or plasma protein or micronutrient levels were uncommon in this cohort of HIV-infected children, even in children with growth retardation. Routine monitoring of the level of proteins and micronutrients studied is unnecessary in the absence of specific clinical indicators of deficiency. PMID- 9404335 TI - Nephrologists' and internal medicine physicians' expectations of renal dietitians and general clinical dietitians. AB - OBJECTIVE: To document and compare nephrologists' and internal medicine physicians' expectations of renal dietitians and general clinical dietitians. DESIGN: Subjects completed a mailed survey. Respondents provided demographic information and used a 5-point Likert scale to note whether each of 14 job functions was appropriate for general clinical dietitians, renal dietitians, or both. SUBJECTS: Five hundred forty-one physicians registered with the Ohio State Medical Board (OSMB) were surveyed. Within this group were 283 nephrologists (the population of nephrologists registered with the OSMB) and 258 internal medicine physicians (selected randomly by the OSMB). A total of 133 physicians (25%) returned the survey; 119 surveys were usable: 70 from nephrologists and 49 from internists. STATISTICAL ANALYSES PERFORMED: A composite variable was created by coding and summing physicians' responses regarding dietitian job functions. This variable was averaged for both physician categories. A t test was conducted to compare composite variable results between the two physician groups. RESULTS: At least 50% of nephrologists and internists agreed that both types of dietitians should conduct nutrition assessments, determine patients' energy needs, evaluate medication-nutrient interactions, recommend diet and tube-feeding orders, instruct patients about physician-ordered diets, and teach nutrition concepts to hospital interns. Few physicians agreed that either type of dietitian should order diets, tube feedings, or diet instructions. APPLICATIONS/CONCLUSIONS: Clinical dietitians can educate physicians about dietitians roles informally in their institutions and formally by supporting programs like The American Dietetic Association Physician Nutrition Education Program. In addition, dietetics educators can hone their students' communication and problem-solving skills to promote positive physician-dietitian interaction. PMID- 9404336 TI - Comparison of dietary risk factors for cardiovascular disease in African-American and white women. AB - OBJECTIVE: To compare African-American and white women's knowledge, attitudes, and energy and nutrient intakes related to cardiovascular disease risk. DESIGN: The 1989 through 1991 Continuing Survey of Food Intakes by Individuals and the Diet and Health Knowledge Survey (DHKS). SUBJECTS: A nationally representative sample of 2,684 white and 449 African-American women who completed the DHKS and provided 3 days of dietary information. STATISTICAL ANALYSES PERFORMED: Comparisons between groups were made using t tests and chi 2 analyses. Analysis of covariance was used to adjust for age, percentage of poverty, and education. RESULTS: Significant differences in dietary risk for cardiovascular disease were identified. White women consumed significantly less cholesterol and more potassium than African-American women. African-American women had significantly lower knowledge scores, but they were more likely than white women to indicate that their diets should be lower in fat and salt and to consider nutrition very important when shopping. White women had more positive attitudes toward the impact of diet on health than African-American women. Accounting for differences between the groups in age, education, and income explained few differences between the groups. APPLICATIONS: This study identified differences in nutrient intake, knowledge, and attitudes about diet and health that can influence the willingness and ability of women to choose foods for a more healthful diet. Dietitians can use these findings to target strategies for changing behavior. For example, many African-American women acknowledge the need to change their current diets, so dietitians can help them design implementation plans or specific action plans to accomplish needed changes. PMID- 9404337 TI - Influence of inhaled corticosteroids and dietary intake on bone density and metabolism in patients with moderate to severe asthma. AB - OBJECTIVES: Compare the effect of high doses of inhaled corticosteroids on bone loss in subjects with moderate to severe asthma or mild asthma, and examine the influence of dietary intake on bone metabolism. DESIGN: A survey on the effects of corticotherapy and nutrition on bone density was conducted in 74 subjects currently being treated for asthma in the asthma clinic of Hopital Laval (Sainte Foy, Quebec, Canada). Fifty-eight subjects completed the study (attrition rate = 15%). MAIN OUTCOME MEASURES: In all subjects expiratory volumes were determined and urinary analysis was conducted for hydroxyproline, calcium, phosphorus, and cortisol levels. Osteocalcin, calcium, phosphorus, cortisol, alkaline phosphatase, and gamma-glutamyltransferase levels were measured in blood samples. Bone density of the lumbar spine was determined by means of dual-energy x-ray absorptiometry. Nutrition evaluation was based on a 3-day food diary analyzed using progiciel Nutri 91. The nutritional parameters examined were calcium; phosphorus; magnesium; zinc; vitamins A, C, and D; protein; total fiber; oxalates; energy; caffeine; and alcohol in relation to bone density. SUBJECTS: Thirty-one patients with moderate to severe asthma who had been taking more than 1,000 micrograms beclomethasone per day or the equivalent for more than 2 years and 27 patients with mild asthma who were taking less than 500 micrograms beclomethasone per day or the equivalent. STATISTICAL ANALYSES PERFORMED: Four factor analysis of variance with hierarchized interactions of four levels, Duncan's test, Pearson correlation coefficients. RESULTS: Blood levels of osteocalcin and protein intake were lower in patients with moderate to severe asthma than in those with mild asthma (P < .05). Significant correlations (P < .02) were observed between bone density and calcium intake (r = .40), phosphorus intake (r = .35), protein intake (r = .30), and serum alkaline phosphatase level (r = -.30). Bone density was not significantly different between the two groups of patients with asthma. APPLICATIONS: A follow-up of patients with asthma who are taking inhaled corticosteroids is needed to assess bone density, osteocalcin levels, and dietary intakes of calcium. Verify if osteocalcin level decreases over time in patients with moderate to severe asthma, monitor possible modifications in bone density, and verify if the correlation between dietary calcium and bone density is maintained. PMID- 9404338 TI - Derivation of daily values used for nutrition labeling. AB - Daily Values (DVs) are the daily dietary intake standards used for nutrition labeling. Information on the derivation of DVs is important for dietetics professionals and nutrition educators who use DVs to educate and instruct patients and students about diet planning and evaluation and about adherence to modified diets. The first daily intake standards for nutrition labeling were established in 1973 and were referred to as the US Recommended Daily Allowances (US RDAs). They were based on the 1968 Recommended Dietary Allowances (RDAs) developed by the National Academy of Sciences. These intake standards were mandatory for 8 and optional for 12 food components on nutrition labels. Regulations revising the daily intake standards for nutrition labeling were published in 1993. The new standards included Reference Daily Intakes (RDIs) for 19 food components and Daily Reference Values (DRVs) for 8 food components. The RDIs were based on the 1973 US RDAs, and the DRVs were based on consensus recommendations. On the nutrition label, the RDIs and DRVs are referred to as DVs. Percent DVs are mandatory on nutrition labels for 10 food components and optional for 16 food components. In 1995, DVs were established by regulation for 6 additional food components; these DVs are optional for nutrition labels. The DVs established in 1995 were based on information from the 1980 and 1989 revisions of the RDAs and Estimated Safe and Adequate Daily Dietary Intakes. Currently, percent DVs are mandatory on nutrition labels for 10 food components and optional for 22. Optional percent DVs become mandatory if claims are made about the food components or if the food components are added to the food through fortification or as food additives. PMID- 9404339 TI - Do oats belong in a gluten-free diet? AB - Celiac disease is an intolerance to protein fractions in wheat, rye, barley, and possibly oats. When these grains are consumed by a person with celiac disease, they damage the mucosa of the small intestine, which eventually leads to malabsorption of nutrients. Patients are therefore advised to remove these grains from their diet, with lifelong adherence generally suggested. Although many dietitians and physicians consider this dietary prescription to be standard protocol, it is actually quite controversial. Whether oats can safely be consumed by persons with celiac disease has been debated since the gluten-free diet was first advocated more than 40 years ago. Historically, there have been several reasons for this debate, including the difficulty in identifying the precise amino acid sequence in gliadin that is responsible for toxicity; the differences in cereal chemistry between wheat and oats; and the lack of well-designed studies to assess the toxicity of oats. A growing body of evidence now suggests that moderate amounts of oats may be safely consumed by most adults with celiac disease. If further research continues to find no adverse effects from oat consumption, a consensus may emerge on the place of oats in the gluten-free diet. In the meantime, individual dietary prescriptions, routinely assessed for appropriateness using histologic and/or serologic studies, may be warranted to prevent unnecessary dietary restrictiveness and undesirable medical complications. PMID- 9404340 TI - Nutrition management of congenital glucose-galactose malabsorption: a case study. AB - In this article we describe the clinical history, diagnostic evaluation, and management of an infant who had congenital glucose-galactose malabsorption (CGGM) -a rare disorder thought to be inherited as an autosomal recessive trait. Because of defective sodium-coupled cotransport of glucose and galactose in the intestinal mucosa, infants with CGGM suffer from chronic, profuse, watery diarrhea that often leads to hypertonic dehydration. This infant experienced persistent diarrhea and hypernatremic dehydration during the first 3 months of life. Despite management with elemental formulas and continuous nasogastric feedings during initial hospitalizations, worsening diarrhea and dehydration persisted and malnutrition occurred. Diagnostic evaluations ruled out cystic fibrosis and bacterial or viral gastroenteritis. Diagnostic tests also revealed normal pancreatic exocrine function and normal villus architecture. The persistence of glucose-positive, watery diarrhea, even when the infant was fed an oral electrolyte solution, led to the diagnosis of CGGM. The infant was treated successfully with a carbohydrate-free infant formula to which fructose was added incrementally to meet energy requirements. Parental education about dietary management of CGGM with specialized formula supplemented with fructose and solid food feedings was an important component of this infant's nutrition therapy. Aggressive nutrition intervention for the infants and judicious dietary counseling for parents can lead to normal growth and neurological development for an infant with CGGM. PMID- 9404341 TI - Evaluation of nutrition care provided to patients with traumatic injuries at risk for multiple organ dysfunction syndrome. PMID- 9404342 TI - Heights and weights of Head Start preschool children in Hawaii. PMID- 9404343 TI - Position of the American Dietetic Association: management of health care food and nutrition services. PMID- 9404344 TI - [Conn's syndrome. Study of systemic cardiovascular hemodynamics after medical treatment with spironolactone and after surgical treatment]. AB - OBJECTIVES: Assess hemodynamic effect of Conn's syndrome in order to better prepare patients for surgical resection of their adenoma. PATIENTS AND METHODS: Hemodynamic investigations were conducted before any treatment in 13 patients with Conn's syndrome. Results were compared with those in 13 control subjects with permanent primary hypertension. In the 13 patients with Conn's syndrome, the same hemodynamic parameters were studied in 13 after drug therapy using spironolactone and in 8 after surgery. RESULTS: Hypertension was associated with a significant increase in stroke volume and a non-significant increase in cardiac index. Blood pressure normalized after sprironolactone and after surgery in parallel with a significant decrease in blood volume. DISCUSSION: These hemodynamic disorders in Conn's disease patients suggest that the increase in stroke volume is a consequence of increased venous return and more likely, in myocardium contractility or a combination of both. CONCLUSION: These hemodynamic characteristics of Conn's disease should be useful in guiding monitoring schemes for these patients in the perioperative period. PMID- 9404345 TI - [Malaria attacks after returning from endemic areas. Failure or inadequate chemoprophylaxis?]. AB - OBJECTIVE: Determine the causes of malaria attacks in subjects who have returned from endemic areas by assessing prescriptions for chemical prophylaxis and compliance. PATIENTS AND METHODS: All patients who developed a paroxysmal episode of malaria diagnosed at the University of Nice hospital in 1995 answered specific questions concerning their anti-malaria prophylaxis. RESULTS: Thirty-three patients were hospitalized for paroxysmal episodes of malaria in 1995. In 32 cases (97%) the attack resulted from either the lack of any prophylaxis (17 cases, 52%), inadequate prescription (11 cases, 12%) or poor compliance (4 cases, 12%). The prescribed chemical prophylaxis was not adapted to the chloroquinone resistant area in 8 cases (24%) and medical recommendations concerning administration rules were inadequate in 3 cases (9%). Only one patient developed a paroxysmal episode despite correct compliance to a chloroquine-resistant zone adapted well-conducted prescription. The cost of poor prophylaxis in terms of human suffering and financial cost was high for this preventable disease. Four patients had to be hospitalized in the intensive care unit and one died during hospitalization. The cumulative cost of hospitalization for these 33 cases was evaluated at 660,000 FF. CONCLUSION: Preventive measures for malaria must include better information for physicians on changing recommendations for chemical prophylaxis as well as better information for travelers provided by all those involved in organizing travel to endemic areas. PMID- 9404346 TI - [Poisoning with "poppers", a rare cause of methemoglobinemia observed in emergency cases]. AB - BACKGROUND: Methemoglobulinemia should be entertained as a differential diagnosis in patients with cyanosis. Recently in France there has been an increase in the number of cases of acquired methemoglobulinemia due to inhalation of poppers. CASE REPORTS: Four patients were admitted to the emergency room of a Paris hospital in a state of unconsciousness with cyanosis. All four patients had inhaled poppers shortly before admission. The clinical course was rapidly favorable after intravenous infusion of methylene blue in 3 cases. DISCUSSION: Poppers are inorganic aliphatic nitrites used for their relaxing effect on smooth muscle and for their aphrodisiac effect. One poorly recognized effect is the development of methemoglobulinemia. Tissue hypoxia results because methemoglobulin cannot bind oxygen, leading to a brown or blue coloration of the blood. Methemoglobulin usually results from exposure to a wide variety of oxidizing compounds including certain drugs. Methylene blue is the specific treatment for symptomatic methemoglobulinemia. These four cases emphasize the toxic effect of products sold in sex shops and calls attention to the life threatening risks involved. PMID- 9404347 TI - [Type 12 F Streptococcus pneumoniae meningitis in a splenectomized patient: failure of vaccination]. PMID- 9404348 TI - [Epididymitis in Behcet disease]. PMID- 9404349 TI - [At the National Academy of Medicine. Desire for pregnancy in the couples with a HIV-seronegative woman and a seropositive partner. A report presented by R. Henrion]. PMID- 9404350 TI - [Value of techniques of molecular biology in the management of hepatitis C virus infections]. PMID- 9404351 TI - [Treatment of malignant hemopathies with all transretinoic acid]. PMID- 9404352 TI - [Indications for thoracoscopy]. PMID- 9404353 TI - [Well-being of insulin-dependent diabetics. Evaluation of 100 adolescents and young adults in relation to their metabolic control]. AB - OBJECTIVE: The principal aim of therapeutic management of the child, adolescent and adult with type I diabetes is to avoid severe hypoglycemia and long-term complications, by maintaining blood glucose concentrations and thus glycated hemoglobin levels (HbA1c)-dose to the normal range. However, the therapeutic constraints should not decrease the quality of life and well-being of patients. Therefore, the purpose of the present study was to evaluate by a questionnaire the well-being of our autonomous diabetic adolescents and young adults in relationship with their HbA1c levels and other characteristics. PATIENTS AND METHODS: A total of 100 unselected subjects (73 men and 44 women), with a mean age of 21 years (14-38) and a mean diabetes duration of 12 years (0-26), were included in the study over a 3-month period. Mean age at onset of diabetes was 10 years. Twenty-five percent of the patients were of Moroccan origin. All the patients were autonomous for self-management and treatment. Their socioeconomic status was not different from that of the normal population. The mean annual HbA1c level in the 100 diabetic patients was 7.3 (4.7-11.7). Well-being was measured using a questionnaire developed by a working group of the World Health Organisation, International Diabetes Federation and St Vincent Declaration. The questionnaire included 4 subscales labelled depression, anxiety, energy and positive well-being. The measurement of all 4 subscales involved 22 items and allowed an estimation of general well-being. RESULTS: General well-being in women was not as good as in men due to a greater tendency toward depression. Well being was better in patients with a professional activity than in the others. Patients age, duration of diabetes, number of insulin injections, frequency of home blood glucose monitoring, presence of 1 or 2 subclinical complications, had no effect on well-being. On the other hand, well-being was negatively correlated with the HbA1c levels: higher the HbA1c, higher the anxiety and the depression, and lower the energy and the positive well-being. CONCLUSION: Well-being was mainly associated with HbA1c levels; it improved with better glucemic control. PMID- 9404354 TI - [Blood levels of homocysteine in patients under 55 years of age with acute coronary insufficiency]. AB - OBJECTIVES: High blood levels of homocysteine have been recently described as a risk factor for thromboembolic events and early development of atherosclerosis. The aim of this work was to study homocysteine blood levels in patients under 55 years of age with acute coronary artery disease. PATIENTS AND METHODS: The study included 110 patients (98 men, 12 women) with poorly controlled angina pectoris (n = 35) or in the acute phase of myocardial infarction (n = 65). Homocysteine was assayed by liquid chromatography in all patients on the day of the acute episode and 24 hours later. Homocysteine levels were also determined in 40 controls under 55 years of age with no history of coronary artery disease. RESULTS: Blood level of homocysteine was 10.6 +/- 6.2 mumol/l in the patients and 7.7 +/- 2.5 mumol/l in the controls (p < 0.01). The difference was greater in the 30-40 year age rang with 14.4 +/- 2 mumol/l in patients versus 6.4 +/- 1.5 mumol/l in controls (p < 0.001). The assays were reproducible at 24 hours (difference less than 10%). The levels were significantly higher in patients with several diseased arteries than those with single-artery disease. The difference between patients and controls was especially remarkable for non-smokers and those with high cholesterol levels. CONCLUSION: Hyperhomocysteinemia would be a factor favoring early development of coronary atherosclerosis. PMID- 9404355 TI - [Renal fibromuscular dysplasia and cerebral aneurysm in a hypertensive patient with a familial history of cerebral vascular complications]. AB - BACKGROUND: The possibility of an association between two vascular anomalies, renal artery dysplasia and cerebral artery aneurysm, merits recognition. CASE REPORT: We report the case of a 63 year old woman who was found to have fibromuscular dysplasia affecting the right renal artery while being investigated for systemic hypertension. Given a family history of cerebrovascular accident occurring before the age of 50, a cerebral angiogram was performed which demonstrated a saccular aneurysm of the middle cerebral artery measuring 6 mm in diameter. DISCUSSION: The association of these two anomalies could result from a familial arterial fibromuscular dysplasia. There are practical implications, notably the risk of aneurysm rupture and the role of hypertension. PMID- 9404356 TI - [True and false osteomalacia: diagnostic value of bone histo-morphometrics]. PMID- 9404357 TI - [Crohn disease disclosed by cerebral infarction. Favoring role of protein C deficiency]. PMID- 9404358 TI - [Local complications after intravenous injection of dissolved tablets of buprenorphine]. PMID- 9404359 TI - [What degree of glycemic control can be obtained in diabetics?]. AB - In diabetic patients, blood glucose should be controlled to a level which prevents acute metabolic complications, forestalls the development of micro and macroangiopathic complications and remains compatible with good quality of life. Recent interventional trials in both insulin-dependent and non-insulin-dependent patients have helped identify this target glucose level. Maintaining HbA1c between 7 and 7.5% appears to be a realistic objective with minimum risk of severe hypoglycemia. This goal, which may be difficult to reach in certain patients, can only be achieved through the co-ordinated efforts of patients and health care providers. The ideal system includes patient education, renewed training for general practitioners who care for most of the non-insulin-dependent as well as a large number of insulin-dependent diabetic patients in France, close follow-up with regular consultations (calling upon specialists when therapeutic adaptations are required) and an organized system of nursing care by specially trained caregivers. The extra cost of this combined organization is to be balanced against expenditures for complications of diabetes. PMID- 9404360 TI - [Neuropsychology, what is the use?]. PMID- 9404361 TI - [Analgesics: the WHO standards and degree of efficacy]. PMID- 9404362 TI - [Anal incontinence: role of perineal neuropathy]. PMID- 9404363 TI - [Fractures of the proximal femur. Epidemiology and economic impact]. PMID- 9404364 TI - [Fractures of the proximal femur. Risk factors]. PMID- 9404365 TI - [Human recombinant granulocytic growth factors: glycosylation makes the difference]. PMID- 9404367 TI - Ischaemia/reperfusion, inflammatory responses and acute lung injury. PMID- 9404366 TI - Salbutamol enantiomers: early clinical evidence in humans. PMID- 9404368 TI - Obstructive sleep apnoea: a progressive disorder? PMID- 9404369 TI - Effect of single doses of S-salbutamol, R-salbutamol, racemic salbutamol, and placebo on the airway response to methacholine. AB - BACKGROUND: Commercially available salbutamol is a racemic mixture consisting of equal amounts of the two enantiomers, R-salbutamol and S-salbutamol, felt to be active and inert, respectively. METHODS: A double blind, randomised, four way, crossover study was performed in 12 well controlled asthmatic subjects (forced expiratory volume in one second (FEV1) > 70% predicted, no beta 2 agonists for > or = 4 weeks). Subjects were studied on four days at intervals of 48 hours to seven days. FEV1 was assessed before and both FEV1 and methacholine PC20 were measured 20 and 180 minutes after a single dose of nebulised racemic salbutamol 2.5 mg, R-salbutamol 1.25 mg, S-salbutamol 1.25 mg, and placebo. RESULTS: Equivalent bronchodilation was seen for both R-salbutamol and racemic salbutamol (mean (SE) 12.4 (3.1)% and 12.0 (3.0)%, respectively, at 20 minutes and 5.9 (2.9)% and 5.2 (2.2)% at 180 minutes). The increase in FEV1 of 5.2 (0.9)% at 20 minutes and the decline in FEV1 of 2.9 (2.1)% at 180 minutes after S-salbutamol were not significantly different from the placebo response. Compared with placebo the methacholine PC20 after R-salbutamol and racemic salbutamol improved by 3.3 (95% CI 2.5 to 4.1) and 3.4 (95% CI 2.6 to 4.2) doubling doses, respectively, at 20 minutes and 1.2 (95% CI 0.6 to 1.8) and 1.0 (95% CI 0.2 to 1.8) doubling doses at 180 minutes. S-salbutamol resulted in an improvement of 0.9 (95% CI 0.3 to 1.5) doubling doses at 20 minutes and no change at 180 minutes. Restlessness (n = 11) and increased pulse were seen 20 minutes after racemic and R-salbutamol but not S-salbutamol or placebo, and not at 180 minutes. There were no other adverse events. CONCLUSION: A single dose of 1.25 mg nebulised R-salbutamol produced equivalent bronchoprotection, bronchodilation, restlessness, and tachycardia as did 2.5 mg of racemic salbutamol. S-salbutamol 1.25 mg had a weak bronchoprotective effect; this could be because of a small amount of contamination with R-salbutamol or because S-salbutamol is an intrinsically weak beta 2 receptor stimulant. PMID- 9404370 TI - Pharmacokinetics and extrapulmonary beta 2 adrenoceptor activity of nebulised racemic salbutamol and its R and S isomers in healthy volunteers. AB - BACKGROUND: Racemic salbutamol remains one of the most commonly used bronchodilators in the treatment of reversible airways obstruction. Data from animal and human studies suggest that the S-isomer, whilst contributing no bronchodilator activity, may induce increased bronchial hyperreactivity and may explain the adverse effects of regular racemic salbutamol on asthmatic disease control. The purpose of this study was to evaluate the dose-response effects of racemic (+/-) salbutamol and its R(-) and S(+) isomers in terms of pharmacokinetics and pharmacodynamics at extrapulmonary beta 2 adrenoceptors when given by the inhaled route to healthy volunteers. METHODS: Twelve healthy volunteers of mean age 20.6 years were studied in a double blind, placebo controlled, crossover design comparing cumulative doubling doses of nebulised R salbutamol (R) and S-salbutamol (S) isomers (200 micrograms/400 micrograms/800 micrograms/1600 micrograms/3200 micrograms) and racemic salbutamol (RS) (400 micrograms/800 micrograms/1600 micrograms/3200 micrograms/6400 micrograms). Doses were administered at 20 minute intervals (t0/t20/t40/t60/t80) and measurements were made of extrapulmonary beta 2 responses as an increase in finger tremor and heart rate and fall in plasma potassium at baseline and each dose level (t0/t20/t40/t60/t80/t100). Plasma levels of salbutamol were measured at 15 minutes after each dose with a further sample at 30 minutes after the last dose (t110). RESULTS: Pharmacodynamics showed dose related beta 2 responses for R salbutamol and RS-salbutamol but not for the S isomer, and a plateau in response was not reached within the administered dose range. No differences in responses were found between R-salbutamol and RS-salbutamol when compared on a 1:2 microgram basis. The effects of the S isomer were indistinguishable from those of placebo. For all beta 2 responses there were differences between R-salbutamol and S-salbutamol (for t100 response as change from placebo); tremor (log units): R 0.74 vs S 0.03 (95% CI 0.39 to 1.03); fall in potassium (mmol/ 1): R 0.35 vs S 0.02 (95% CI 0.03 to 0.71). Pharmacokinetics showed consistently higher levels for S-salbutamol than R-salbutamol at 15 minutes after each dose, with R salbutamol already being cleared and S-salbutamol reaching peak levels at 30 minutes after the last dose (at t110). There were higher plasma levels of R salbutamol and S-salbutamol following administration of the respective isomers alone compared with their levels after administration of the racemate, suggesting an influence of each isomer on the clearance of the opposite isomer when given as a racemate. CONCLUSIONS: The S-isomer of salbutamol has no detectable activity at extrapulmonary beta 2 adrenoceptors whilst exhibiting higher plasma levels than the R-isomer, in keeping with greater clearance of R-salbutamol than S salbutamol. Inhalation of R-salbutamol and RS-salbutamol produced dose-related beta 2 responses which were equivalent when compared on a 1:2 microgram basis, despite higher plasma levels of R-salbutamol after administration of the R isomer than after administration of the racemate. Further dose ranging studies are required at steady state to evaluate the pharmacokinetics of R- and S-salbutamol and their relative effects on bronchial hyperreactivity when given on a regular basis to asthmatic subjects. PMID- 9404371 TI - Cross sectional investigation of the effects of inhaled corticosteroids on bone density and bone metabolism in patients with asthma. AB - BACKGROUND: Bone mineral density has been reduced in patients with asthma taking inhaled corticosteroids in some cross sectional studies and this could be important if treatment is continued for several decades. The possibility of confounding by age, menopausal status, physical activity and, especially, past oral steroid use has not been excluded in most studies. The present study was designed to assess the magnitude of any reduction in bone mineral density in relation to inhaled steroid use after adjusting for these factors. METHODS: Bone mineral density (BMD), vertebral fractures, and markers of bone metabolism (serum osteocalcin, procollagen peptide I, bone-specific alkaline phosphatase, and urinary deoxypyridinoline cross links) were measured in 81 patients with asthma age 20-40 years; 34 patients (19 men) who had never had inhaled or systemic steroids and 47 (19 men) who had taken inhaled steroids for at least five years with limited exposure to systemic steroids in the past. Data relating to past medication use, physical activity, smoking, and other confounding factors were collected by questionnaire. The relation between inhaled steroid dose and duration and BMD was assessed by linear regression analysis, accounting for potential confounders including weight, exercise, and oral steroid use. RESULTS: The 47 patients taking an inhaled steroid had a mean current dose of 620 micrograms/day (range 100-3000 micrograms), a mean duration of use of 7.8 years, and had had a mean of 0.85 courses of prednisolone in the past. There was no significant difference in mean BMD values between those who were and those who were not on inhaled steroids in men or women. However, on multivariate analysis, cumulative inhaled steroid dose was associated with a reduction in posterior anterior (P-A) and lateral lumbar spine bone mineral density in women, equivalent to a 0.11 standard deviation reduction in bone density per 1000 micrograms/day inhaled steroid per year after adjustment for potential confounding factors (95% CI for P-A spine 0.01 to 0.22; for lateral spine 0.02 to 0.21). Previous oral steroid use was not an important confounding factor in these patients. Inhaled steroid use was not related to BMD at the wrist or hip in women or at any skeletal site in men. Women taking an inhaled steroid had lower levels of serum osteocalcin than those not taking them, although this was not dose related. Inhaled steroid use was not associated with differences in other markers of bone metabolism in men or women or with the presence of vertebral fractures. CONCLUSIONS: Although an effect of confounding factors cannot be excluded entirely in a cross sectional study, our findings are in keeping with an effect of inhaled steroid therapy in reducing bone density in the spine in women and provide an estimate of the magnitude of this effect. PMID- 9404372 TI - Attenuation of propranolol-induced bronchoconstriction by frusemide. AB - BACKGROUND: Inhaled propranolol causes bronchoconstriction in asthmatic subjects by an indirect mechanism which remains unclear. Inhaled frusemide has been shown to attenuate a number of indirectly acting bronchoconstrictor challenges. The aim of this study was to investigate whether frusemide could protect against propranolol-induced bronchoconstriction in patients with stable mild asthma. METHODS: Twelve asthmatic subjects were studied on three separate days. At the first visit subjects inhaled increasing doubling concentrations of propranolol (0.25-32 mg/ml), breathing tidally from a jet nebuliser. The provocative concentration of propranolol causing a 20% reduction in FEV1 (PC20FEV1 propranolol) was determined from the log concentration-response curve for each subject. At the following visits nebulised frusemide (4 ml x 10 mg/ml) or placebo (isotonic saline) was administered in a randomised, double blind, crossover fashion. FEV1 was measured immediately before and five minutes after drug administration. Individual PC20FEV1 propranolol was then administered and FEV1 was recorded at five minute intervals for 15 minutes. Residual bronchoconstriction was reversed with nebulised salbutamol. RESULTS: Frusemide had no acute bronchodilator effect but significantly reduced the maximum fall in FEV1 due to propranolol: mean fall 18.2% after placebo and 11.8% after frusemide. The median difference in maximum % fall in FEV1 within individuals between study days was 3.6% (95% CI 1.2 to 11.7). CONCLUSIONS: Frusemide attenuates propranolol induced bronchoconstriction, a property shared with sodium cromoglycate. Both drugs block other indirect challenges and the present study lends further support to the suggestion that frusemide and cromoglycate share a similar mechanism of action in the airways. PMID- 9404373 TI - Acute lung injury after aortic surgery: the relation between lung and leg microvascular permeability to 111indium-labelled transferrin and circulating mediators. AB - BACKGROUND: Aortic surgery is a risk factor for acute lung injury and this may relate to ischaemia/reperfusion (I/R) of the lower body and release of inflammatory mediators. The aim of this study was to define the changes in microvascular protein permeability and circulating inflammatory mediators after aortic surgery. METHODS: In 11 consecutive patients who underwent elective aortic surgery microvascular permeability in lung and leg was measured before and a median of 2.8 hours after completion of surgery using 111indium (In)-labelled transferrin and 99mtechnetium (Tc)-labelled red blood cells, yielding a protein leak index (PLI) that is specific for protein permeability. Circulating leucocyte counts and levels of inflammatory mediators were determined. RESULTS: In the lung the PLI rose from a median of 0.6 (range -0.5 to 2.2) x 10(-3)/min before surgery to 5.4 (-2.3 to 33.5) x 10(-3)/min after surgery, and in the leg from 0.3 (-1.6 to 1.7) x 10(-3)/min to 5.0 (1.0 to 27.8) x 10(-3)/min. The increase in PLI in the lung was related to that in the leg. Levels of activated complement C3a and tumour necrosis factor-alpha did not change, but levels of interleukin (IL)-6, IL 8 and elastase-alpha 1-antitrypsin increased. After surgery there was slight neutrophilia and the leucocyte counts were inversely related to the IL-8 level. The rise in lung but not in leg PLI was greatest in patients with the highest IL 8 levels and the lowest leucocyte counts. CONCLUSIONS: Early after aortic surgery microvascular protein permeability increases in the leg and lung. Leg I/R injury may result in neutrophil activation and release of IL-8, which may induce neutrophil sequestration and subsequently increased pulmonary microvascular permeability. These findings may help to explain the occurrence of acute lung injury after I/R in man. PMID- 9404374 TI - Natural evolution of moderate sleep apnoea syndrome: significant progression over a mean of 17 months. AB - BACKGROUND: Obstructive sleep apnoea (OSA) is associated with increased morbidity and mortality. It has remained unclear whether or not it is progressive. The evolution of OSA was examined in a retrospective case note study of 55 unselected patients of mean (SD) age 55.8 (10) years with mild to moderate disease untreated by interventional methods such as continuous positive airway pressure (CPAP) or surgery. Correlations between clinical and functional variables, upper airway anatomy, and change in disease severity were also investigated. METHODS: Patients underwent full polysomnography on two occasions (T0 and Tx) at a mean interval of 77 (50) weeks (range 17-229). In addition, upper airway imaging with computed tomographic scanning or cephalometry had been performed in 43 patients at T0. Morbidity before, during, and after the study period was assessed by questionnaire, as was smoking history and alcohol and sedative intake. RESULTS: The apnoea hypopnoea index (AHI) for the group as a whole increased from 21.8 (11.5) to 33.4 (21.3) (p = 0.0001). Using a 25% change in AHI to divide patients into worsened, stable, and improved groups showed that, although most of the patients deteriorated, 25 patients improved or remained stable. The change in AHI was not correlated with body mass index which remained stable at 29.7 (5.4) kg/m2 versus 29.7 (5.6) kg/m2. There was a trend for apnoea duration to increase. No patient reported increased alcohol consumption and only one patient reported increased use of sedatives between T0 and Tx. No correlation was found between change in AHI and age, time between recordings, anatomical measurements of the upper airway, respiratory function, oximetry, or arterial blood gas tensions. Total cardiovascular and cerebrovascular morbidity was high: hypertension (26 patients, 46%), cardiac arrhythmia (17 patients, 33%), angina (12 patients, 23%), myocardial infarction (10 patients, 19%), and stroke (10 patients, 19%). Twenty nine patients (52%) were prescribed CPAP after Tx, two of whom went on to have maxillofacial surgery. These 29 treated patients had significantly higher values of AHI at T0 and Tx and greater change in AHI than the untreated patients. CONCLUSIONS: This study shows that mild to moderate OSA has a tendency to worsen in the absence of significant weight gain and that upper airway anatomy and clinical variables do not appear to be useful in predicting progression. It follows that mild to moderate OSA justifies systematic follow up. Deterioration in AHI over a mean of 17 months led to interventional treatment in over 50% of patients in the study. PMID- 9404375 TI - Comparison of outcome measures for patients with chronic obstructive pulmonary disease (COPD) in an outpatient setting. AB - BACKGROUND: To assist clinicians and researchers in choosing outcome measures for patients with chronic obstructive pulmonary disease attending routine outpatient clinics, a comparative assessment was undertaken of four questionnaires designed to reflect the patients' perception of their physical and emotional health in terms of their feasibility, validity, reliability, and responsiveness to health change. METHODS: Two condition specific questionnaires, the St George's Respiratory Questionnaire (SGRQ) and Guyatt's Chronic Respiratory Questionnaire (CRQ), and two generic questionnaires, the Short Form-36 Health Survey (SF-36) and Euroqol (EQ), were compared for their discriminative and evaluative properties. Spirometric tests and a walking test were also performed. One hundred and fifty six adults who were clinically judged to have COPD and who attended an outpatient chest clinic were assessed at recruitment and six and 12 months later. Patients were also asked whether their health had changed since their last assessment. RESULTS: Completion rates and consistency between items for dimensions of the SGRQ were lower than for dimensions of the other questionnaires. The distributions of responses were skewed for certain dimensions in all questionnaires except the CRQ. Validity was supported for all instruments insofar as patients' scores were associated with differences in disease severity. The generic questionnaires better reflected other health problems. All instruments were reliable over time. The condition specific questionnaires were more responsive between baseline and first follow up visit but this difference did not persist. While certain dimensions of the SF-36 were responsive to patient perceived changes, this did not apply to the derived single index of the EQ. The rating scale of the EQ, however, provided a quick and easy indicator of change. CONCLUSIONS: Evidence from this study supports the CRQ and the SF-36 as comprehensive outcome measures for patients with longstanding COPD. PMID- 9404376 TI - Cryptogenic fibrosing alveolitis: lack of association with Epstein-Barr virus infection. AB - BACKGROUND: Cryptogenic fibrosing alveolitis (CFA) is a well defined clinical entity of unknown aetiology. An association between CFA and the presence of protein indicating Epstein-Barr virus (EBV) replication within epithelial cells of the respiratory tract has recently been suggested, leading to speculation for a role for EBV in the pathogenesis of CFA. METHODS: Lung tissue was obtained from patients in three groups: those with cryptogenic fibrosing alveolitis, either lone or associated with systemic sclerosis; patients with other pulmonary disorders; and patients with normal lung. Paraffin blocks were stained using three antibodies raised against well defined EBV antigens. In addition, EBER-1 and EBER-2 anti-sense nucleotide probes were used in an attempt to identify EBV RNA. DNA was also extracted from the tissue sections and evaluated for evidence of EBV DNA using the polymerase chain reaction. RESULTS: Immunohistochemistry showed inconsistent focal positive staining with anti-EBV antibodies in all three groups, but there was no evidence of EBV RNA using in situ hybridisation. None of the samples from patients with pulmonary fibrotic disorders was found to contain EBV DNA following gene amplification. CONCLUSION: Contrary to an earlier report, these results do not support the hypothesis that EBV has a role in the pathogenesis of CFA. PMID- 9404377 TI - Risk of malignant neoplasms in patients with pulmonary sarcoidosis. AB - BACKGROUND: For over 20 years the association between sarcoidosis and malignancy, particularly lymphoma and lung cancer, has been disputed with misclassification being the major concern. The aim of the present study was to analyse the incidence of malignancies in a cohort of patients with sarcoidosis by linkage to a nationwide population based cancer register. METHODS: The cohort comprised 254 patients followed for a median of 25 years until death, emigration, or 31 December 1992, whichever came first. The expected number of cancer cases was calculated using the annual age and sex specific cancer rates from the Danish Cancer Registry. RESULTS: Thirty six cancers were registered, three of which were misclassified as sarcoidosis, leaving 33 cancers compared with 23 expected (standardised incidence ratio (SIR) = 1.4; 95% CI 0.99 to 2.0). Five lung cancers were observed compared with 2.5 expected, yielding an SIR of 2.0 (95% CI 0.7 to 4.7). There was no incidence of lymphoma and only one case of leukaemia. There was a significant excess number of pharyngeal cancers based on two cases (SIR = 15.4; 95% CI 1.7 to 56). CONCLUSIONS: This study does not support the theory of an association between sarcoidosis and malignancy, and the main reason other studies have shown such an association is most likely to have been due to selection bias and misclassification. PMID- 9404378 TI - Relation of fetal growth to adult lung function in south India. AB - BACKGROUND: Follow up studies in Britain have shown that low rates of fetal growth are followed by reduced lung function in adult life, independent of smoking and social class. It is suggested that fetal adaptations to undernutrition in utero result in permanent changes in lung structure, which in turn lead to chronic airflow obstruction. India has high rates of intrauterine growth retardation, but no study has examined the association between fetal growth and adult lung function in Indian people. We have related size at birth to lung function in an urban Indian population aged 38-59 years. METHODS: Two hundred and eighty six men and women born in one hospital in Mysore City, South India, during 1934-1953 were traced by a house-to-house survey of the city. Their mean forced expiratory volume in one second (FEV1) and forced vital capacity (FVC) were measured using a turbine spirometer. These measurements were linked to their size at birth, recorded at the time. RESULTS: In both men and women mean FEV1 fell with decreasing birthweight. Adjusted for age and height, it fell by 0.09 litres with each pound (454 g) decrease in birthweight in men (95% confidence interval (CI) 0.01 to 0.16) and by 0.06 (95% CI -0.01 to 0.13) in women. Likewise, mean FVC fell by 0.11 litres (95% CI 0.02 to 0.19) with each pound decrease in birthweight in men, and by 0.08 litres (95% CI 0.002 to 0.16) in women. FEV1 and FVC were lower in men who smoked, but the associations with size at birth were independent of smoking. Small head circumference at birth was associated with a low FEV1/FVC ratio in men which may reflect restriction in airway growth in early gestation. CONCLUSION: This is further evidence that adult lung function is "programmed" in fetal life. Smoking may be particularly detrimental to the lung function of populations already disadvantaged by poor rates of fetal growth. PMID- 9404379 TI - Effect of increasing doses of hypertonic saline on mucociliary clearance in patients with cystic fibrosis. AB - BACKGROUND: Patients with cystic fibrosis are known to have decreased mucociliary clearance. It has previously been shown that inhalation of a 7.0% solution of hypertonic saline significantly improved mucociliary clearance in a group of adult patients with cystic fibrosis. The aim of this study was to measure the response to increasing concentrations of inhaled hypertonic saline. METHODS: Ten patients (seven men) of mean (SE) age 22 (4) years and mean forced expiratory volume in one second (FEV1) 52.0 (6.7)% predicted completed the study. Mucociliary clearance was measured using a radioaerosol technique for 90 minutes after the interventions which comprised 0.9% NaCl + voluntary cough (control), 3.0% NaCl, 7.0% NaCl, and 12% NaCl. RESULTS: There was a significant increase in the amount of activity cleared from the right lung with all concentrations of hypertonic saline (HS) compared with control. The amount cleared at 90 minutes on the control day was 12.7% (95% confidence interval (CI) 9.8 to 17.2) compared with 19.7% (95% CI 13.6 to 29.5) for 3% HS, 23.8% (95% CI 15.9 to 36.7) for 7% HS and 26.0% (95% CI 19.8 to 35.9) for 12% HS. The improvement in mucociliary clearance was not solely due to coughing as the number of coughs recorded on the control day exceeded that recorded on any other day. The hypertonic saline did not induce a clinically significant change in FEV1. CONCLUSIONS: Within the range of concentrations examined in this study, the effect of hypertonic saline appears to be dose dependent. Inhalation of hypertonic saline remains a potentially useful treatment for patients with cystic fibrosis. PMID- 9404380 TI - Health effects of passive smoking. 1. Parental smoking and lower respiratory illness in infancy and early childhood. AB - BACKGROUND: A systematic quantitative review was conducted of evidence relating parental smoking to acute lower respiratory illness in the first three years of life. METHODS: Fifty relevant publications were identified after consideration of 692 articles selected by electronic search of the Embase and Medline databases using keywords relevant to passive smoking in children. The search, completed in April 1997, identified 24 studies ascertaining illnesses in a community setting, including five surveys of schoolchildren with retrospective ascertainment of early chest illness, and 17 studies of admissions to hospital for lower respiratory illness in early life. Thirty eight studies were included in a quantitative overview using random effects modelling to derive pooled odds ratios. RESULTS: The results of community and hospital studies are broadly consistent, with only one publication reporting a reduced risk among children of smokers. The pooled odds ratios were 1.57 (95% CI 1.42 to 1.74) for smoking by either parent and 1.72 (95% CI 1.55 to 1.91) for maternal smoking. There is a significantly increased risk of early chest illness associated with smoking by other household members in families where the mother does not smoke (1.29, 95% CI 1.16 to 1.44). The associations with parental smoking are robust to adjustment for confounding factors, and show evidence of a dose-response relationship in most studies in which this has been investigated. CONCLUSIONS: The relationship between parental smoking and acute lower respiratory illness in infancy is very likely to be causal. Although it is impossible to distinguish the independent contributions of prenatal and postnatal maternal smoking, the increased risk associated with smoking by other household members suggests that exposure to environmental tobacco smoke after birth is a cause of acute chest illness in young children. PMID- 9404381 TI - Erythromycin and diffuse panbronchiolitis. PMID- 9404382 TI - Adenosine bronchoprovocation: a promising marker of allergic inflammation in asthma? PMID- 9404384 TI - Three cases of meningococcal pneumonia. AB - Three cases of pneumonia due to Neisseria meningitidis are described. In all three cases the organism was isolated only from blood cultures, but in the presence of good clinical and radiological evidence of pneumonia. The isolates belonged to three different serogroups: B type 2b, C, and Y. The cases illustrate the fact that N meningitidis can cause pneumonia and that culture of blood plays an important part in the diagnosis. Clinically there is nothing to differentiate meningococcal pneumonia from other causes of community acquired pneumonia. Predisposing factors include aspiration, immunosuppression, influenza, and adenovirus infections. When diagnosed, pneumonia due to N meningitidis should be notified and prophylaxis given as for meningitis or septicaemia. PMID- 9404383 TI - Proteoglycans: the "Teflon" of the airways? AB - Proteoglycans are a family of structurally distinct, polyanionic complex carbohydrates composed of repeating disaccharide units. Proteoglycans include heparin, heparan sulphate, chondroitin 4-sulphate, chondroitin 6-sulphate, dermatan sulphate, and hyaluronic acid. Heparin is found in the granules of a subset of mast cells where it is bound to various mediators including histamine. Heparan sulphate has a much wider distribution in the body, being associated with stromal matrices, basement membrane and many cell surfaces, particularly the surface of endothelial cells. Heparin is an anticoagulant, but it is now very apparent that it possesses many other biological activities that have relevance to our understanding of lung diseases, particularly inflammatory diseases of the airway. Recent evidence suggests in the airway when administered by inhalation that could be exploited therapeutically. PMID- 9404385 TI - Markers of inflammation in induced sputum in acute bronchitis caused by Chlamydia pneumoniae. AB - Little is known of the inflammatory characteristics of acute infections of the respiratory tract caused by virus and unusual bacteria such as Chlamydia pneumoniae. A case is reported in whom inflammatory indices in sputum were used to investigate, for the first time, the airway inflammation during an episode of acute bronchitis caused by C pneumoniae. The patient presented with a dry cough of five days duration. C pneumoniae was identified by polymerase chain reaction (PCR) in a nasopharyngeal swab collected on day 5. Virological studies were negative. Clinical and inflammatory indices in induced sputum were measured on days 6, 8, and 11. The cough cleared spontaneously by day 11. Forced expiratory volume in one second was normal throughout. Sputum findings identified intense airway inflammation characterised by increased total cell and lymphocyte counts followed by an increase in neutrophils and a decrease in the CD4/CD8 ratio, activation of CD8 lymphocytes, and exudation as indicated by an increase in fluid phase fibrinogen. These observations suggest that sputum might be useful to monitor an inflammatory/immune response of the airway in acute infections. PMID- 9404386 TI - Increase in incidence of childhood empyema. PMID- 9404387 TI - Asthma guidelines. PMID- 9404388 TI - Births, marriages, divorces, and deaths for January 1997. PMID- 9404389 TI - Asian or Pacific Islander mortality, selected states, 1992. AB - OBJECTIVES: In this report the National Center for Health Statistics (NCHS) presents mortality data in greater race detail than has previously been presented for the Asian or Pacific Islander (API) population. METHODS: Deaths, estimated death rates, age-adjusted death rates, exploratory life expectancies, and ranking of leading causes are presented for a selected area. RESULTS: In 1992 a total of 19,478 deaths occurred in the Asian or Pacific Islander population in the seven States examined in this report. Heart disease and cancer were the two leading causes of death for each of the Asian or Pacific Islander subgroups. By age there is striking variation in leading causes among the race groups. Among the API subgroups in these States, age-adjusted death rates are greatest for the Samoan and Hawaiian populations and smallest for the Asian Indian, Korean, and Japanese populations. Life expectancy is lowest for the Hawaiian and Samoan populations. PMID- 9404391 TI - Births, marriages, divorces, and deaths for February 1997. PMID- 9404390 TI - Births and deaths: United States, 1996. AB - OBJECTIVES: This report presents preliminary data on births and deaths in the United States from the National Center for Health Statistics for 1996. U.S. data are shown by age, race, and Hispanic origin. National and State data on births by marital status, prenatal care, cesarean delivery, and low birthweight are also presented. Mortality data presented include life expectancy, leading causes of death, and infant mortality. METHODS: This report, the third in a new statistical series, presents preliminary data for 1996 on births and deaths based on a substantial sample of vital records. The records are weighted to independent control counts of births, infant deaths, and total deaths received in State vital statistics offices during calendar year 1996. RESULTS: According to preliminary data for 1996, the birth rate for teenagers dropped 4 percent in 1996 to 54.7 births per 1,000 women aged 15-19 years. The teen birth rate has declined 12 percent since 1991 (62.1), with larger reductions for young teenagers 15-17 years and for black teenagers. Birth rates for women aged 20-34 years increased 1-2 percent, while rates for women aged 35-44 years rose 3 percent. The number and percent of births to unmarried women increased about 1 percent, while the birth rate for unmarried women declined 1 percent. The rate of prenatal care utilization improved and the cesarean delivery rate declined. The overall low birthweight rate increased to 7.4 percent. The 1996 preliminary infant mortality rate reached a record low of 7.2 infant deaths per 1,000 live births with all time lows for white and black infants. Life expectancy reached a record high of 76.1 years with all-time highs for white and black males and black females. The largest declines in age-adjusted death rates among the leading causes of death were for Human immunodeficiency virus (HIV) (26 percent) and Homicide (11 percent), which dropped from the 12th to the 14th leading cause of death. PMID- 9404392 TI - Births, marriages, divorces, and deaths for March 1997. PMID- 9404393 TI - Quality of life in HIV-infected women in the south-eastern United States. AB - The purpose of this study was to evaluate the relative importance of social (social support, material resources, disclosure, and family functioning) and psychological factors (stigma, emotional distress, intrusion, avoidance, and fatalism) as predictors of the quality of life of women infected with HIV. The cross-sectional data were drawn from interviews of a sample of 264 women recruited from 8 HIV/AIDS treatment sites in a south-eastern state. Variance in quality of life variables, included limited daily functioning, general anxiety, and HIV symptoms was analyzed using ANOVA, correlations, and hierarchical multiple regression analysis. Limited daily functioning was predicted by stigma, fatalism, employment status, and stage of disease (R2 = 0.179). General anxiety was predicted by emotional distress, intrusion, and marital status (R2 = 0.503). Reported HIV symptoms were predicted by material resources, disclosure, intrusion, age, employment status, and race (R2 = 0.294). The results of this study support that social and, particularly, psychological factors are important in their influence on quality of life in women with HIV infection and suggest the need for interventions which address such factors. PMID- 9404394 TI - A survey of women with HIV about their expectations for care. AB - As an increasing number of women become infected with the human immunodeficiency virus (HIV), it is important to understand their expectations regarding the health care services they receive. In 1995 a new centre was opened at an academic centre in St Louis, Missouri, to provide comprehensive care to women with HIV. To assist in the Centre's development, we interviewed 50 of 119 enrolled clients (42%) using a survey instrument focusing on what they consider important in their care. In response to open-ended questions, clients most often mentioned wanting a sense of personalized caring and respect by medical staff (n = 28, [56%]), having someone to talk to about problems (n = 28, [56%]), honest answers about their condition (n = 23, [46%]), medical follow-up (n = 21, [42%]), reduced barriers to care (n = 20, [40%]), and education about their condition (n = 15, [30%]). The highest-ranked aspects of care were seeing the doctor, learning about their condition, and being seen in a pleasant environment (92% [n = 46%]). Significant differences were found in some responses when analysed according to race, educational level, and severity of disease. It is important that programmes delivering health care services to women with HIV provide services that take into account their individualized needs. Ideally, this requires incorporation of a multidisciplinary team to provide psychological and social support, patient education, and medical management. PMID- 9404395 TI - Trauma of discovery: women's narratives of being informed they are HIV-infected. AB - The purposes of this research report are to describe women's subjective experiences of being informed of a positive HIV antibody test and, from their point of view, to explain the meaning and impact of discovering that one is HIV infected. In this qualitative narrative study, a racially diverse, low-income sample of 38 HIV-infected women shared their stories of HIV discovery during in depth interviews. Findings of a multi-staged narrative analysis suggest that, for women like those in this study, the discovery of HIV seropositivity is a traumatic event, carrying with it elements that are common to other types of trauma: perceived threat to one's life and perceived responsibility for the deaths of others. Overarching personal meanings, or metaphors, framed these women's experiences of the trauma of HIV discovery. HIV discovery was an epiphany for 10% of the sample, a confirmation for 37%, and a calamity for 53%. Among their calamitous reactions were shock, fear, anguish, and suicidality. The impact of learning that they were HIV-infected often took its toll in unrelenting misery, escalated drug use, transmission risks, and destabilization of relationships, income, and shelter. Extensive excerpts from participants' interviews illustrate analytic findings. Implications for counselling and follow up at diagnosis and early in the course of HIV illness are elaborated. PMID- 9404396 TI - Vulnerability on the streets: female sex workers and HIV risk. AB - In-depth interviews were conducted with 24 purposively selected female sex workers who were perceived to be vulnerable to risks associated with their lifestyle and occupation. Brothel workers were found to be considerably less exposed to risk than the women working on the streets. Client resistance was the major obstacle to women maintaining safe sex practices. Physical threats and coercion from clients, the absence of legal protection for street workers, the workers' extreme social isolation and lack of community support added to the difficulties experienced by women in their attempts to insist on condoms for all sex services. Youth, homelessness and heavy drug use had contributed to women being at times even more vulnerable because they had less capacity to manage situations of potential violence or STD risk. Whether through sex work or in their private relationships, HIV remains a risk for some of these women. This study highlights the dangers associated with illegal sex work. While decriminalization of prostitution would reduce some of the dangers to which women were exposed and increase women's capacity to insist on safe sex practices, it is also important for community education programmes to address men's failure to accept responsibility for condom use when seeking the services of sex workers. PMID- 9404397 TI - 'Appropriate' male and female safer sexual behaviour in heterosexual relationships. AB - This paper is an attempt to explore how traditional gender role socialization runs counter to safer sexual practices. Structured interviews (N = 448 sexual encounters) were conducted with heterosexual young adults in 'natural settings' such as summer music festivals and youth houses/youth clubs. Congruent with a perspective of gender role theory, females were found to care more about safer sex. The data do not suggest that females are forced to rely on less efficient ways of practising safer sex, but they have to try harder to be successful. Males may implement safer sex in an encounter without negotiation, because it is obvious, or they just have to mention it at the moment of sexual intercourse. Females have to start negotiating safer sex long before the actual encounter in order to be as successful as males. There is one important critical situation for females, notably the encounter in which they perceive the male to be dominant. PMID- 9404398 TI - Concomitants of HIV/STD risk behaviours and intention to engage in risk behaviours in adolescents in India. AB - A total of 1230 year 11 and 12 anglophone college students, modal age 16 and 17, in three colleges in Mumbai (Bombay), India, were studied with regard to sexual behaviours or risk of sexual behaviours, beliefs about sex, HIV/STD knowledge, and perceived norms regarding sexual behaviours. Data indicated that 8% of males and 1% of females had had sexual experience, but over one-third were not sure at all of being able to abstain from sexual activity with either steady or casual partners. However, perceived norms were slanted toward sexual abstinence for the majority of the sample. Knowledge of the protective effects of condoms was high, although half of those who had had sex did not use condoms. Logistic regression showed that knowledge was higher among males, those who believed it was OK to have sex with a steady partner and that they should not wait until they were older, those who believed that condoms should be used even if the partner is known, and those who believed it was acceptable to have multiple partners. Gender differences in sexual activity and beliefs about sexual activity showed that males were less likely to believe in abstaining from sexual activity and to engage in it. We conclude that this age-group is appropriate for HIV/STD reduction education given the low rate of sexual activity but that, despite knowledge of the importance of condom use, the social skills to apply this knowledge are lacking. PMID- 9404399 TI - Prevention of vertical HIV transmission with zidovudine: projected impact of HIV testing and prenatal care. AB - We sought to estimate the impact of maternal HIV testing and prenatal care on the potential to reduce vertical transmission through zidovudine (AZT) use by HIV infected mothers. We evaluated the prepartum maternal HIV diagnosis rate, prenatal care, disease stage, and vertical transmission rate (from a two-part mixture model) using New York State Medicaid and vital statistics data for HIV infected mothers and their singletons in 1985-90. We used published data to estimate the effect of AZT on vertical transmission and expert input to define other parameters for the model. Our HIV-infected (N = 1514) had a vertical transmission rate of 27.0%. HIV was diagnosed prepartum for 39.5% of women in 1990. Transmission would have been 22.2% if AZT had been taken only by the subset of women diagnosed prepartum with HIV and receiving prenatal care by 34 weeks gestation (86.7%). Transmission would have dropped to 11.2% if all women had been diagnosed prepartum with HIV and received adequate prenatal care. The observed deficiencies in prenatal care and maternal HIV diagnosis rates in this Medicaid population-based cohort must be addressed to realize the promise of AZT to reduce vertical transmission. PMID- 9404400 TI - Disclosing HIV status and sexual orientation to employers. AB - Seropositive gay and bisexual men who reveal their sexual orientation or HIV/AIDS status to their employer risk discriminatory reprisals. However, non-disclosure may limit potential social, emotional, and tangible support. Among our sample of 389 seropositive gay and bisexual men employed in the US, 52% were 'out' to their employer and 35% had disclosed their serostatus to him or her. Among gay men, employer awareness of their sexual orientation was related to their being European American (vs Latino or African American), being HIV-seropositive for more than 4 years, and having a gay or bisexual employer. Disclosure of HIV infection in the total sample was related to being European American, HIV seropositive for more than 4 years, symptomatic (vs asymptomatic), 'out' at work, and having a gay or bisexual employer. Men who had informed their employers of their HIV status reported consequences that were substantially more positive than those anticipated by men who had not disclosed. Policy and research implications for improving the work environment for gay and bisexual men living with HIV are considered. PMID- 9404401 TI - Caring for patients with HIV disease: the experience of a generic hospice. AB - This study describes the experience of a generic hospice admitting people with advanced HIV disease over a 4-year period. Data were collected retrospectively for all patients with HIV disease admitted. The aim of the study was to review the number of referrals, the reason for referral, subsequent symptom control and multidisciplinary team involvement together with the outcome for these patients. Twenty-six patients were admitted for the first time. Two patients were female, 24 were male; median age was 36 years (range 25-58 years). Hospitals referred more patients than general practitioners (18 (70%) and 5 (20%) respectively), but most were from non-HIV specialist areas within hospitals (11 (42%)). The commonest reason for referral was locality, particularly in terms of ease of access. The most prevalent symptoms on admission were weakness, immobility and weight loss (77%, 73% and 62% respectively). These were not improved during admission. There was significant improvement in the control of other symptoms including pain, gastrointestinal disturbance, confusion and dyspnoea. Use of the full multidisciplinary team was high. Median length of stay was 19 days (range 1 77 days). Seventeen patients (65%) died on their first admission. This study confirms the high prevalence of symptomatology among patients with HIV disease. Many generic hospices can offer skilled multidisciplinary symptom control and psychosocial care, complementing other HIV specialist services. It is important that patients with HIV disease and specialist health care professionals working in the HIV field are made aware of what generic hospices are able to offer so that patients can make informed choices about their care. PMID- 9404402 TI - Parental HIV discordancy and its impact on the family. AB - There is little or no available information on the effect of HIV discordancy in heterosexual relationships on different family members. A review of case notes was carried out on all families who had a child referred to the paediatric HIV service/family clinic at St Mary's Hospital between January 1991 and March 1996. The children had been exposed to HIV infection because they were born to HIV positive women. There was HIV discordancy in more than one-fifth of the parents' relationships. In over 46% of the relationships, the HIV status of the natural or birth father was not known because he was either untested or unavailable. It is likely that not all of these men are infected and the number of discordant couples is greater. There were more discordant couples where the man and woman came from different ethnic groups. Consideration of the potential impact of discordancy on individual men, women and children is discussed. PMID- 9404403 TI - [Ventriculoarterial coupling and left ventricular performance in hypertensive patients with left ventricular hypertrophy]. AB - It has been shown that 1) contractile performance of hypertrophied left ventricle (LV) of hypertensive patients (HP) is depressed, and 2) ventriculoarterial (VA) coupling is altered when myocardial contractile performance is reduced and when afterload is increased. To assess the relationship between contractile performance of hypertrophied LV and the VA coupling in hypertensive patients. LV angiography coupled with simultaneous recording of pressures with micromanometer were used to determine end-systolic stress/volume index ratio (ESS/ESVi), the slope of end-systolic pressure-volume relationship, i.e. end-systolic elastance (Ees), effective arterial elastance (Ea), external work (EW) and pressure-volume area (PVA). Comparison of results in 30 HP and 20 control subjects (CS) showed that LV contractile performance assessed by Ees/100 g left ventricular myocardial mass (LVM, echocardiographic determination) was depressed (HT: 4.35 +/- 1.13; CS: 5.21 +/- 1.89 mmHg/ml/100 g; p < 0.02) and was negatively correlated to the LVM (Ees = -0.026 LVM + 3.363; r = 0.581; p < 0.001), when ESS/ESVi, another estimate of LV contractile performance, was comparable in the 2 groups (6.66 +/- 1.55 g/cm2/ml/m2 in HT vs 6.72 +/- 1.36 in CS; NS) and negatively correlated with the LVM (ESS/ESVi = -0.019 LVM + 8.947; r = 0.369; p < 0.01). Ventriculoarterial coupling evaluated through Ea/Ees ratio (Ea and Ees in mmHg/ml/m2) was slightly higher in HT (0.53 +/- 0.08 vs 0.48 +/- 0.09 in CS; p < 0.05), work efficiency (EW/PVA) was similar in the 2 groups (0.78 +/- 0.04 in HP vs 0.80 +/- 0.03 in CS) and PVA, which is representative of the myocardial oxygen demand per beat, is negatively related to LVM (PVA = -0.003 MVG + 1.44; r = 0.434; p < 0.01). CONCLUSIONS: this study shows that despite a slight depression of LV contractile performance, work efficiency is preserved and ventriculoarterial coupling is almost normal in HP with LV hypertrophy. Thus, it appears that LV hypertrophy might be a useful means of preservation of matching LV and arterial receptor with minimal energetical cost. PMID- 9404404 TI - [Left ventricular repercussion of obesity-induced arterial hypertension in the dog]. AB - Obesity and hypertension are frequently associated. The aim of our study was to assess the effects of high fat diet on weight, blood pressure and left ventricule in dogs. We studied 6 male Beagle dogs before and after 7 weeks of hypercaloric hyperlipidic diet. Echocardiography was used to measure left ventricular wall thickness, volumes, ejection fraction and mass. Results are expressed as % of variation of initial values. After 20 weeks, dogs presented abdominal obesity with increased body weight (11.9 +/- 2.3 to 15.2 +/- 2 kg; p < 0.03) associated with an increasing of systolic (196.5 +/- 14.6 to 260.1 +/- 17.5 mmHg; p < 0.03), diastolic (76.6 +/- 9 to 110.6 +/- 10.2; p < 0.004) and mean blood pressure (128.8 +/- 7 to 152.7 +/- 7.6 mmHg; p < 0.004). There were non significant changes concerning diastolic thickness of septum and posterior wall. Left ventricular volumes increased in diastole (41.1 +/- 4.5 to 48.9 +/- 10.3 cm3; p < 0.03) and systole (12.2 +/- 1.7 to 14.9 +/- 3.2 cm3; p < 0.03). So, despite any changes in wall thickness, we observed an increased of ventricular mass (67 +/- 15 to 80 +/- 24.3 g; p < 0.03). Ejection fraction remained unchanged. CONCLUSION: it appears that hight fat diet induces obesity and hypertension in dogs; changes in left ventricule suggest a volodependent hypertension. PMID- 9404405 TI - [Response of the coronary arteries to cold test and flow velocity increase is improved by deferoxamine but not by L-arginine in diabetic patients]. AB - Acetylcholine produces coronary artery (CA) constriction in diabetic patients suggesting an impairment of endothelium-dependent dilation. To examine the mechanism of this abnormal response. 2 physiological tests, i.e. cold pressor test (CPT) and coronary flow-increase induced by 10 mg papaverine (PAP) injection in the distal left anterior descending CA (dLAD), were performed before (1) and after (2) either i.v. L-arginine (L-arg, 625 mg/min x 10 min) or i.v. desferrioxamine (DFX, 50 mg/min x 10 min) in 15 normotensive nonsmoker diabetic patients with angiographically normal CA and normal cholesterol. Dimensions of the proximal LAD (pLAD) were measured by quantitative angiography. [table: see text] Before administration of L-arg or DFX, CPT induced a decrease of pLAD diameter, and PAP injection in dLAD dit not modify pLAD diameter. In the 7 diabetic patients receiving L-arg, responses to CPT and PAP were not modified. Conversely in the 10 patients receiving DFX, pLAD dilated in response to the 2 tests. Intracoronary isosorbide dinitrate, an endothelium-independent dilator, produced similar dilation in the 2 groups (+20 +/- 8% and +16 +/- 6%, respectively). CONCLUSIONS: 1) responses of angiography normal CA to CPT and to flow increase are impaired in diabetic patients; 2) abnormal responses are not improved by L-arg suggesting that a deficit in substrate for NO synthesis is not involved; 3) DFX restores a vasodilator response to the 2 tests suggesting that inactivation of NO by superoxide radicals might be partly responsible of the impairment of CA dilatation in diabetic patients. PMID- 9404406 TI - [Determinants of the left ventricular mass in obese patients. Influence of lean body mass]. AB - Systolic blood pressure and body mass index (BMI) are the main determinants of the left ventricular mass (LVM). The mechanism of this cardiac hypertrophy in the obese individual is multifactorial and involves hemodynamic as well as metabolic factors. The association of LVM with the morphologic features of the individual are well known. The aim of this study was to assess the influence of the morphologic and metabolic features of obese women on LVM. 2D echocardiography evaluation of LVM was done in 24 normotensive, normoglycemic obese women (BMI [27.5-52.2 Kg/m2). Lean and fat body mass were determined by bio-impedancemetry, insulin sensitivity (Si) by the minimal model (Bergman), and basal metabolism by using indirect calorimetry. There was a positive correlation between LVM and BMI (r = 0.61; p = 0.001), waist to hip ratio (r = 0.45; p = 0.03), basal metabolism (r = 0.61, p = 0.001), lean (r = 0.74, p = 0.0002) and fat (r = 0.49; p = 0.01) body mass. Fasting glycemia was positively correlated with LVM (r = 0.62; p = 0.001), but not Si. LVM was also positively correlated to the triglyceride level. No relations were found with systolic or diastolic blood pressure. Multivariate regression analysis was performed to determine the relative contribution of lean body mass (the morphologic variable with the best association to LVM in univariate analysis), blood glucose, waist to hip ratio, age and triglycerides. The multiple r for the model was 0.87 (p < 0.001). Lean body mass and blood glucose were found to be the only significant and independent predictors of LVM (p = 0.001 and p = 0.03 respectively). We conclude that: 1) lean body mass is an important determinant of LVM in obese normotensive individuals. Hence, in obese women, correcting LVM for lean body mass might be more accurate than correcting it for body surface area or height. 2) There is no relationship between LVM and insulin sensitivity. The link between blood glucose and LVM needs to be studied further. PMID- 9404407 TI - [Prognostic value of ventricular arrhythmia in hypertensive patients]. AB - OBJECTIVE: Hypertensive left ventricular hypertrophy (LVH) is associated with increased risk of arrhythmias and mortality. However, no clinical study demonstrated a significant relation between ventricular arrhythmias and mortality in systemic hypertension. DESIGN AND METHODS: To evaluate the prognostic value of arrhythmogenic markers in systemic hypertension, we included between 1987 and 1993. 214 hypertensive patients, 59.1 +/- 12.8 years old, without symptomatic coronary disease, myocardial infarction, systolic dysfunction, electrolyte disturbances or antiarrhythmic therapy. At inclusion, an ECG, a 24 h Holter ECG (204 patients) with Lown classification of ventricular arrhythmias, an echocardiography (reliable in 187 patients) with left ventricular mass index and ejection fraction calculation, a SAECG (125 patients, enrolled after 1988) with ventricular late potentials (LP) were recorded. QT interval dispersion (QTd) was calculated on 12 leads standard ECG and LVH was appreciated. RESULTS: At baseline echocardiographic LVH was recorded in 63 patients (33.7%) with normal ejection fraction (75 +/- 7.4%). Non-sustained ventricular tachycardia (Lown IVb) was found in 33 pts (16.2%) and LP in 27 patients (21.6%). After a mean follow up of 42.4 +/- 26.8 months, all-cause mortality was 11.2% (24 patients); 17 patients died of cardiac causes (7.9%); of these 9 patients (4.2%) died suddenly. In univariate analysis, age, strain pattern of LVH, advanced Lown classes and abnormal QT dispersion (> 80 ms) were significantly related to global, cardiac and sudden death (p < or = 0.01). Left ventricular mass index was closely related to cardiac mortality (p = 0.002). LP failed to predict mortality. In multivariate analysis, only Lown class IVb was an independent predictor of global and cardiac mortality, increasing the risk of global death 2.6 fold [1.2-6.0] (CI 95%) and the risk of cardiac death 3.5 fold [1.2-9.7] (CI 95%). CONCLUSIONS: In hypertensive patients the presence of non-sustained ventricular tachycardia on 24 h Holter has a prognostic value. PMID- 9404408 TI - [Value of scintigraphy using meta-iodo-benzyl-guanidine (MIBG) in the investigation of cardiac autonomic neuropathy in diabetic patients. Comparison with Ewing tests]. AB - Cardiac autonomic neuropathy is a frequent complication of diabetes leading to resting tachycardia, postural hypotension, painless myocardial ischaemia, rhythm disturbances and sudden cardiac death. The aim of the study was to evaluate in a diabetic population the sensitivity of two exploration modes of autonomic neuropathy in diabetics: the Ewing tests which are, at present time, the reference method and the (123-I) meta-iodo-benzyl-guanidine (MIBG) single photon emission computed tomography (SPECT) which evaluates the cardiac sympathetic innervation. PATIENTS AND METHODS: 9 male insulin-dependent diabetes mellitus patients were studied. Mean age was 40.7 +/- 15 years and diabetes duration was 10.8 +/- 6 years. None had hypertension or macroangiography as demonstrated by patient's history, clinical examination, rest and exercise electrocardiography and ambulatory blood pressure monitoring. The complications observed were background retinopathy in 2 patients, incipient nephropathy in 3 and a peripheral neuropathy in 1 patient. Ewing tests, i.e. Valsalva maneuver, beat to beat heart rate variation during deep breathing and standing, blood pressure response to standing and to sustained handgrip, were performed. The results were considered as pathologic when the score was over 2. After injection of 10 mCi (123-I) MIBG, planar images were realized at times 1, 2 and 4 hours and SPECT images after 2 hours. The heart/mediastinum uptake ratio was calculated. RESULTS: We noted abnormalities of planar images in 3 patients, SPECT images in 1, and both in 1 patient. None was positive for Ewing tests. CONCLUSION: Although MIBG SPECT will explore only the sympathetic innervation, these preliminary findings suggest that this technique could be more sensitive for the evaluation of cardiac autonomic neuropathy. Nevertheless cost and lack of disponibility of this technique should limit its use. PMID- 9404409 TI - [Prevalence and profile of renovascular disease in type II diabetic patients with severe hypertension]. AB - The aim of this study was to determine the prevalence and profile of renal artery stenosis (RAS) in NIDDM population with severe hypertension. 60 consecutive NIDDM with severe HT (> or = 3 hypotensive drugs), 42 F/18 M (SR: 2.8), mean age: 66.6 +/- 6.5 years, diabetes duration: 14.1 +/- 6 years have had metabolic, ABPM and renal investigations: color duplex scan (CDS) (with renal us): n = 60, and/or arteriography: n = 17). 13 (21.5%) renal artery stenosis > or = 70%: 8 unilateral/5 bilateral were proved by arteriography. We compared classic HT (n = 47) versus renovascular HT (n = 13). There was no difference for age (years): 64.8 +/- 8 versus 70.6 +/- 6.4, HT duration (years): 11.6 +/- 6.8 versus 12.3 +/- 6. B.M.I.: 31.5 +/- 6 versus 27.6 +/- 3.3, HBA1C (%): 8.9 +/- 2.2 versus 8.8 +/- 0.9, cholesterol (mmol/L): 5.7 +/- 1.3 versus 5.5 +/- 0.6. Significant difference (p < 0.05) was noticed for S.R. (F/M): 2.9 versus 1.16, diabetes duration (years): 11.7 +/- 5 versus 16.5 +/- 8, frequency of retinopathy (%): 30 versus 61, smoking (%): 10 versus 40, triglycerides (mmol/L): 1.9 +/- 1.1 versus 2.6 +/- 1.1, and (p < 0.01) for blood pressure level (mmHg) (SBP: 142 +/- 20 vs 155 +/- 7, DBP: 81 +/- 13 vs 87 +/- 10, MBP: 103 +/- 16 vs 111 +/- 6), frequency (%) of HT escape (> or = 140/SBP, > or = 90/DBP) on ABPM: 40 versus 75 and 24 versus 40, insulin requirence (%): 36 versus 69, macroangiopathy (%): 51 versus 100 (coronaropathy: 34 vs 61, legs arteritis: 21 vs 69, carotid stenosis: 17 vs 30) and for renal function: frequency (%) of micro-macroalbuminuria: 36 versus 92 creatinaemia (mmol/L): 80 +/- 24 versus 124 +/- 44, creatinaemia clearance (mmL/min): 65 +/- 30 versus 40 +/- 12 while are found 5 renal insufficiencies (> or = 120 mmol/L). In NIDDM population with severe HT, renovascular HT is frequent (21.5%), and RAS must be evocated in unstable HT and/or renal injury with macro angiopathy, old NIDDM (> 15 years), requiring insulin. Colour duplex scan (+ renal US) mays lead to arteriography to confirm renal artery stenosis. PMID- 9404410 TI - [Prevalence and aspects of arteriopathies in non-insulin-dependent diabetes mellitus with severe hypertension]. AB - Severe hypertension may lead to macroangiopathy complications especially when a major vascular risk factor as diabetes exists. We have studied the prevalence of macroangiopathy in a group of 40 consecutive NIDDM patients with severe hypertension (> or = 3 hypotensive drugs) (grS) that we have compared to 80 consecutive NIDDM patients with controlled hypertension (1 or 2 hypotensive drugs) (grC). All patients have had metabolic, blood pressure (ABPM) and vascular (color duplex) investigations. The two groups were similar for age (years): 61.9 > or = 9 versus 65.2 +/- 9.5, diabetes duration (years): 10.7 +/- 7 versus 12.1 +/- 8 and hypertension duration: (years) 8.9 +/- 8 versus 11.7 +/- 7.3. The mean level of blood pressure was the same in all patients (mmHg): SBP = 138 +/- 14 versus 144 +/- 20; DBP = 80 +/- 9 versus 83 +/- 13; MBP = 100 +/- 10 versus 105 +/- 15. The frequency (%) of escape SBP (> 140): 50 versus 80, p < 0.01), and DBP (> 90): 29 versus 35, p < 0.05 was significantly higher in grS. Twenty (25%) patients in grC and 20 (50%) in grS had one or more macroangiopathy which was dispatched as follow: coronary heart disease n = 8 (7%) versus 13 (32.5%), p < 0.01; lower limb arteritis n = 12 (15%) versus n = 9 (22%), NS; carotid atheroma n = 5 (25) versus n = 6 (15%), NS. All significant renal artery stenosis (RAS) n = 8 (20%) were found in grS (p < 0.001). Only plasma triglyceride level (mmol/L) was statistically higher in grS 2.5 +/- 1.2 versus +/- 1 while BMI, plasma cholesterol, HbA1C, and creatininemia were NS. The sex-ratio (F/M) 1.28 versus 3, insulin requirement (%): 11 versus 42.5, retinopathy (%) 14 versus 45 and micromacroalbuminuria were statistically significant p < 0.01. CONCLUSION: macroangiopathy is frequent in severe hypertension (50%) versus controlled hypertension (25%) in NIDDM patients especially coronary heart disease (32.5%); the prevalence of RAS is high in grS (20%). The following criteria are frequently noticed in high risk patients: insulin requirement, micro or macroalbuminuria and high plasma triglyceride. PMID- 9404411 TI - [Decrease of vascular response to iloprost in diabetic rats]. AB - The functional dilatory response in the streptozotocin-induced diabetic rat was investigated using thoracic aortas and coronary microcirculation. The aortas were cut in 4 mm intact or denuded rings and mounted into 20-ml organ baths. Coronary microcirculation was evaluated with isolated hearts perfused under constant flow conditions. Firstly, vasodilation to iloprost (Ilo) was examined. Dose-response curves to Ilo (10 pM-10 microM) on phenylephrine (PE, 30 nM for endothelium denuded, and 0.3 microM for intact) preconstricted rings of diabetics and age matched controls were comparable (n = 6). Decreased vasodilation in diabetic group was observed when dose-response curves to Ilo (1 nM-0.1 microM) were realized in isolated hearts (-22 +/- 3.3% vs -46 +/- 3.9%, n = 6, p < 0.05). Secondly, dose-response curves to forskolin (FSK), an adenylate-cyclase activator, performed in hearts (1 nM-3 microM), and on PE preconstricted rings (10 pM-10 microM) of diabetics and age-matched controls were comparable. Finally, the effect of an activator of ATP sensitive potassium channels (KATP), cromakalim (CMK), was evaluated in coronary circulation (0.3 nM-3 microM) and in aortas (10 pM-10 microM). Decreased vasodilation to CMK was observed in diabetic hearts ( 10.5 +/- 4.3 vs -30.1 +/- 2.8%, n = 6, p < 0.05). In conclusion, under our experimental conditions, diabetes affects selectively the coronary vasodilation to iloprost. This modification of vascular reactivity may be due to a decrease of KATP channels sensitivity but not to a decreased activity of adenylate-cyclase. PMID- 9404412 TI - [Intima media thickness of the carotid artery in white coat and ambulatory hypertension]. AB - Recent epidemiological studies have reported an association between carotid intima media thickness (IMT), ambulatory blood pressure (ABP) and absolute cardiovascular risk. To study the relation between white coat effect and vascular changes in hypertensives (HT), 57 essential HT (office blood pressure (OBP) 152.2 +/- 19.5/93.7 +/- 12.4 mmHg) were recruited (46.4 +/- 11.8 years old, 49 men). After antihypertensive drugs withdrawal, an ABP was performed (Spacelabs 90207). The right common carotid artery IMT 3 cm proximal to the bifurcation was examined by ultrasonography. IMT (0.59 +/- 0.09 mm; Software lotec system) were measured by a reader blinded to the ABP data. White coat hypertension (WCH) was defined by a mean day-time ambulatory BP (d-ABP) lower than the 90th percentile of the distribution of daytime ABP of a normotensive population reported by Verdecchia et al. (131/86 mmHg in women and 136/87 mmHg in men). [table: see text] White coat hypertension was found in 8 from 57 (14%) subjects. IMT was significantly increased in ambulatory HT when compared with white coat HT while age, sex ratio, OBP, smoking status were not different. In stepwise regression age and systolic d ABP were the only determinants of IMT (p < 0.05). In our hypertensive population. ABP appears more closely related to IMT than OBP and IMT in sustained is greater than in white coat hypertensive. PMID- 9404413 TI - [Short-term variability of blood pressure]. AB - The regulation of blood pressure (BP) is traditionally described in terms of homeostasis, and indicates that BP although being continuously perturbed by external stimulations always displays the tendency to come back toward a reference set point. Experimental and clinical studies indicate that these fluctuations occurring around the average present a source of complementary information on the mechanism of cardiovascular control. Recently a wide variety of algorithm and models have been proposed to study the cardiovascular system through new technics of continuous non invasive BP or heart rate (HR) measurements. They give new insites for the evaluation of hypertensive patients and relevance to the understanding of the role of the disorder of the tonic regulation of BP, rather than its short-term variability or reactivity. However, if available data unequivocally indicate that the analysis of variability is a useful tool, the interpretation of those data in clinical trials is not always optimal because there is lot of interaction between BP, HR and other biological signals, and furthermore the use of laboratory data introduces problems to predict what happens on daily life ambulatory conditions. PMID- 9404414 TI - [Blood pressure determination and medical competence]. AB - Hypertension diagnosis depends closely to the blood pressure measurement. The aim of this work is to show whether blood pressure measurement should be done by a beginner or a competent doctor. The blood pressure of 180 patients, (150 females, 30 males) was taken by two physicians. The patients' average age was 51 +/- 11. One of the two physicians was a cardiologist who took all the patients blood pressure. Others where six doctors in training, that is sixth' year students at the faculty of medicine. They took part in this study for a week. The procedure was that the cardiologist and one of the training doctors took the patient's blood pressure at the same time after 15 min rest. We have calculated the average systolic blood pressure and diastolic blood pressure of 720 measures. Then the difference between the cardiologist's measures and those of all the training doctors. After that we have analysed the difference between the average of 120 measures taken by one of the training doctors and the corresponding measurement of the specialist. We have then compared the difference of the 20 measures of every day taken by the training doctor and the ones taken by the specialist. The difference wasn't statistically significant either for the systolic blood pressure or the diastolic blood pressurement. We have studied the evolution of the average of the 20 measurements of every day during the whole week. So, we have noticed that the difference lowers from the first days to the sixth. In the end, we were interested in the last figure of each measurement of blood pressure. The training doctor often gave measurements up to 0 or 5 whereas the specialist gave precise measurements. We have concluded from this work that if experience is needed, the physician has to know the principals and the tricks of blood pressurement. Moreover, the blood pressure variations by "white coat" effect can't be explained by measurements techniques. This effect can be considered as psychic, interactions between doctors and patients. PMID- 9404415 TI - [Reactivity of "white coat" type is associated with reactivity to mental stress]. AB - This study was aimed to compare the white coat effect and the response to a mental stress. 29 subjects, referred for high blood pressure (BP) were included. Systolic BP (SBP) was recorded beat-to-beat with a Finapres device during 3 periods of at least 5 minutes: 1) rest (alone, in lying position); 2) white coat (5 measurements of BP with a standard mercury sphygmomanometer by the same physician); 3) mental stress (version for computer of the Stroop Word Color Conflict Test). A Coarse-graining spectral analysis was performed to compute the power in the low frequency band (PLF: 0-0.150 Hz) and in the high frequency band (PHF: 0.150-0.500 Hz). SBP was 142 +/- 3.7 during the rest period and increased significantly during the white coat (156.7 +/- 3.9 mmHg) and the mental stress (190.7 +/- 4.8 mmHg) periods. These rises of SBP levels were associated with a rise of PLF, significant only during mental stress (11.3 +/- 1.4, 15.7 +/- 3.7, 17.2 +/- 2.4 mmHg2/Hz, during rest, white coat and mental stress periods, respectively). Moreover, a significant correlation (r = 0.76; p < 0.0001) was found between the white coat effect (PAS "white coat"-PAS "rest") and the response to stress (PAS "stress"-PAS "rest"). This work shows that white coat effect is not a specific response but may rather represent an increased reactivity to stress. As it is associated with an increased power in the LF band like the response to stress, this white coat effect may involve an activation of the sympathetic system. PMID- 9404416 TI - [Baseline blood pressure in healthy normotensive volunteers]. AB - The aim of this study was to define the reproducibility and the time required to obtain a stable baseline level of blood pressure (BP) in normotensive volunteers during a phase I trial. Blood pressure was recorded automatically (Dinamap and Marquette monitors) every 5 min during a 2-hour period and manually (Random zero device) at T20, T60 and T120, twice at a one-week interval, under similar study conditions (6 in the morning, 7 in the afternoon) in supine position in 13 normotensive men (aged 20 to 28). The average BP was compared using a 3-way ANOVA (subject, time, week). 1.SBP/DBP decreased significantly (p < 0.001) from one week to the other and SBP, but not DBP, decreased significantly over time up to T75 (p < 0.001): [table: see text] 2. SBP was significantly higher in the morning than in the afternoon during both weeks (p = 0.001). The decrease in SBP with rest was only observed in the morning (p = 0.00001). 3. Reproducibility and change over time and period did not significantly differ between manual and oscillometric methods. The best reproducibility of T75 was obtained with the mean of 3 automatic values (T70, T75, T80). CONCLUSION: in normotensive subjects, BP decreased from one period to the next and with rest. The baseline value of BP was obtained from T75 with the best reproducibility when baseline BP level is defined by 3 automatic values. PMID- 9404417 TI - [Value of a predictive model of ambulatory blood pressure integrating physical activity]. AB - OBJECTIVE: To determine how much of the variations of blood pressure during a 24 hour period could be accounted for by a change in activity and establish a predictive model. MATERIALS AND METHODS: Twenty three healthy subjects (mean age 25 +/- 2 years) were studied. The BP, heart rate (HR), and time of measure (T) were recorded by ambulatory BP monitoring using Spacelabs (4 measures per hour). At each measure the subject noted in a diary the degree of activity on a six level semi-quantitative scale. DATA ANALYSIS: A model was constructed using an analysis of covariance. Different parameters were added in succession to reach a model of the type P: P0 + A + beta + (HR-HR0) + H, were P = predicted systolic pressure, P0 = mean systolic BP over the 24 hours. A variation in systolic BP for activity level, beta = the slope of the regression between systolic BP and HR during activity A, and HR0 the mean HR during this activity. RESULTS: 1) In order to test the model, the values measured in one subject were compared to the predicted values from the model in 22 others. The procedure was then repeated for the other subjects. This common model predicted 41 +/- 21% of fluctuations in BP of the subject analysed with a range of 0 to 66%. 2) In order to refine the individual model two subjects were explored 7 times over 24 h of non consecutive days. The measures of the last recording were compared to the predicted values from the application of the model to the six preceding recordings. The model then predicted 81% and 66% of the BP values of the test day. The mean of the 24 hour individual difference over a one hour period between the measures and its predicted value by the model was 0.13 +/- 4.8 mmHg, and -0.75 +/- 7.7 mmHg. CONCLUSION: This study expresses in a quantitative fashion the importance of the level of activity in the evaluation of the level of ambulatory BP. The introduction of this method of quantification and analysis seems logical in therapeutic trial. The difference in the predictions by the model for some subjects poses the problem of uniform coding of activities and that of the recognition of other events such as stress and dreaming in sleep. PMID- 9404418 TI - [A case of apparent mineralocorticoid excess caused by type 2 11 beta- hydroxysteroid dehydrogenase deficiency]. AB - The syndrome of apparent mineralocorticoid excess is a recessively inherited form of low renin hypertension. The syndrome is characterised by sodium retention and hypervolemia despite low plasma renin activity and aldosterone levels. Patients with this syndrome have mutations in the 11HSD2 gene which encodes the enzyme which normally converts cortisol in the renal tubule to its inactive form, cortisone. The unconverted cortisol is thus able to bind and activate the mineralocorticoid receptor, displacing its usual ligand, aldosterone, causing the apparent mineralocorticoid excess. We have studied a patient with severe hypertension, low renin and aldosterone, and a chronic hypokalemic alkalosis at age 4. The analysis of cortisone, cortisol and their metabolites showed the specific pattern of the apparent mineralocorticoid excess. In serum and urine, there was a dramatic decrease of cortisone and its metabolite, while cortisol and its metabolites were non affected. PMID- 9404419 TI - [Bosentan attenuates the hypertensive effect of angiotensin II in rats]. AB - The influence of nonspecific blockade of endothelin receptors by bosentan (30 mg/kg per day, gavage) was assessed on hypertension induced by infusion of angiotensin II (AngII 200 ng/kg/min sc for 10 days) in rats. Tail-cuff pressure was measured before and every second day of AngII-infusion period. At the end of experiments, mean arterial pressure (MAP, mmHg), cardiac output (CO ml/min/kg body weight) and renal blood flow (RBF ml/min/g kidney weight) were determined (microspheres technique) in conscious rats, and total peripheral and renal vascular resistances were calculated (TPR = MAP/CO and RVR = MAP/RBF). [table: see text] Tail-cuff pressure increased from 126 +/- 4 to 164 +/- 8 mmHg in rats infused with AngII alone whereas it did not change (basal: 132 +/- 3 and final: 135 +/- 3 mmHg: p = NS) when bosentan was coadministered with AngII. At the end of study in conscious rats, the AngII-induced rise in MAP was accompanied by a reduction in CO and RBF and a marked increased in TPR and RVR. In AngII-perfused rats, CO, RBF, TPR and RVR were restored by bosentan to values observed in untreated rats. These results indicate that blockade of endothelin A and B receptors by bosentan prevents the development of AngII-induced hypertension through attenuation of the effect of AngII on vascular tone and suggest that endothelin is an important mediator of the vasoconstrictor action of angiotensin II in rats. PMID- 9404420 TI - [Gene transfer of interferon beta inhibits vascular smooth muscle cell proliferation in vitro and in animal model of arterial injury]. AB - Vascular hypertrophy may increase the blood pressure by its effect on vascular resistance. In this study, adenoviral gene transfer of IFN-beta was analysed in a porcine model of balloon injury to determine whether a secreted growth inhibitory protein might affect the regrowth of vascular smooth muscle cells (VSMC) in vitro and in arteries. An adenoviral vector encoding IFN-beta (ADV-IFN-beta) was constructed by homologous recombination between sub360 genomic DNA, an ADV 5 derivative with a deletion in the E3 region and a porcine IFN-beta expression plasmid. Its antiproliferative effect was analysed using cell proliferation assays, and used in a porcine model of balloon injury. After injury, arteries were immediately transfected with 7 x 10(9) plaques forming units of either ADV IFN-beta or a control E1A deficient adenovirus that does not encode a recombinant protein, ADV-delta E1. The intima/media (I/M) area ratio was determined by quantitative morphometry 21 days after artery injury and gene transfer. Expression of recombinant porcine IFN-beta in VSMC reduced cell proliferation significantly in vitro, and supernatants derived from IFN-beta vector infected cells inhibited VSMC proliferation relative to controls. When introduced into porcine arteries after balloon injury, a reduction in I/M ratio of 30% was found. I/M ratio in the IFN-beta transduced arteries was 0.54 +/- 0.03 vs 0.69 +/- 0.06 in ADV-delta E1 transfected arteries and 0.702 +/- 0.05 in the non-transfected arteries. Gene transfer of an adenoviral vector encoding IFN-beta to VSMC and injured arteries reduced cell proliferation and vascular thickening. This approach is potentially applicable to vascular proliferative diseases. PMID- 9404421 TI - [Direct gene transfer in the rat kidney in vivo]. AB - Gene delivery to the kidney has both experimental and therapeutic potential in hypertension, although the delivery methods, distribution of transgene and subsequent inflammatory response have been poorly characterized. In adult male Sprague-Dawley rats (200 g, n = 26), the left iliac artery was catheterized and a small catheter (Microbore Tygon S-54-HL) was advanced to the origin of the left renal artery. Loops were tied transiently around the aorta and below the renal arterial bifurcation. After flushing the kidney, the renal vein was tied and 500 microL of transfection solution was instilled. After 15 min all the loops were released, the catheter was removed and the left iliac artery ligated. Both replication-defective adenovirus (ADV) constructions used were based on an Ad5 derivative with a partial E3 deletion. Virus ADV-chloramphenicol acetyl transferase (CAT) and ADV-human placental alkaline phosphatase (hpAP), 10(8), 3 x 10(8), 10(9) and 10(10) plaques forming units/mL (pfu/mL), were used respectively to compare the degree of transfection (CAT) and to localize the transgene in the kidney (hpAP), 48 h after transfection. Controls were infused with vehicle. ADV CAT 10(10) pfu/mL induced a gene expression, respectively, 1.4 (NS), 12 (p < 0.001) and 28 (p < 0.001) fold greater than the 10(9), 3 x 10(8) and 10(8) pfu/mL formulations. HpAP staining was located in the juxta-medullary part of the cortex, predominantly in the interstitium. Genetically-modified cells were identified as endothelial cells, mainly in peritubular capillaries but also in efferent arterioles and hilar arteries. Highly efficient gene transfer achieved with ADV-hpAP 10(10) pfu/mL was associated with focal necrosis of the proximal convoluted tubules. No changes were observed with the other viral concentrations. Gene delivery, mediated by a replication-defective ADV, to one rat kidney via the renal artery, induced a dose-dependent gene expression located in endothelial cells in peritubular capillaries. Toxicity was observed only with the highest viral concentration. PMID- 9404422 TI - [Renal vascular responses of bradykinin in the isolated rat kidney]. AB - Kinins, by an autocrine or paracrine hormonal action, are potent modulators of regional vasomotricity. Their effects on the renal circulation are not well defined. The aim of this study was to analyse the renal vascular response induced by bradykinin, to precise the type(s) of receptor involved and to evaluate the contribution of various peptidases in the local catabolism of the kinin. Experiments were performed on the isolated rat kidney, perfused in an open circuit, at a constant flow of 8 mL/min, with a Tyrode's solution. Vasodilator responses were evaluated after renal vascular tone had been restored by a continuous perfusion with prostaglandin F2 alpha. Infusion of bradykinin (0.1-30 nM) induced a concentration-dependent renal vasorelaxation. A maximal response of 39.5 +/- 2.8% (n = 32) reversion of the tone induced by prostaglandin F2 alpha (about 50% of the maximal response induced by acetylcholine on the same kidneys) was obtained at 30 nM. Bradykinin-induced vasodilatation was completely inhibited by HOE 140 (10 nM), a selective bradykinin B2 receptor antagonist. At a supramaximal concentration of 300 nM, bradykinin-induced vasorelaxation was modulated by a concomitant vasoconstriction. A concentration-dependent vasoconstriction was also obtained with desArg9 bradykinin (1-8 microM), a selective agonist of the bradykinin B1 receptor. The inhibition of neutral endopeptidase by phosphoramidon (10 microM) or the inhibition of carboxypeptidase M by MGTPA (10 microM) did not modify the bradykinin-induced renal vasorelaxation. On the other hand, the inhibition of angiotensin I converting enzyme by lisinopril (1 microM) potentiated by about 32% the vasorelaxant response induced by 30 nM bradykinin (52.3 +/- 11.8% relaxation, n = 5, p < 0.05). Present results demonstrate that 1) bradykinin primarily evokes B2 receptor-linked renal vasodilatation, 2) bradykinin B1 receptors appear also to be present on the rat renal vasculature and 3) angiotensin 1 converting enzyme contributes to the local vascular catabolism of the kinin. PMID- 9404423 TI - [Renal tissue angiotensins during converting enzyme inhibition of angiotensin I in spontaneously hypertensive rat]. AB - To compare the effects of an angiotensin-converting enzyme inhibitor on circulating and tissue renin-angiotensin system, we measured different renin angiotensin system parameters during the first day of treatment (Day 1) as well as after two weeks of treatment (Day 14). Ramipril was given orally once daily to adult male spontaneously hypertensive rats. Renin activity, angiotensin converting enzyme activity and levels of angiotensin I and angiotensin II in the plasma, renal cortex and renal medullar were assessed at Day 1 and Day 14 of the treatment. In the plasma, both renin activity and angiotensin I increased 10 to 15 fold one to four hours after acute as well as at Day 14 of ramipril treatment and then returned to basal values within 24 hours. Plasma angiotensin II levels were not significantly decreased at Day 1 or Day 14. The decrease in the angiotensin II/angiotensin I ratio suggested a sustained inhibition of plasma angiotensin-converting enzyme at Day 14. In the renal cortex and medulla, a clearly different pattern was observed: in ramipril treated rats, renin activity in the renal cortex and medulla did not change at Day 1 but at Day 14 we observed a slight and sustained increase in renin activity. Despite very high basal levels of renin activity, angiotensin I levels in the renal cortex were comparable to those in the plasma. The angiotensin I level increased only one-fold one hour after ramipril intake at Day 1 and Day 14. This suggests that angiotensinogen may have a limiting role in the synthesis of angiotensin I in the kidney. Angiotensin II levels were slightly higher in the renal cortex and medulla than in the plasma suggesting local synthesis of the peptide. In the kidney, angiotensin II levels decreased one and four hours after the acute or prolonged ramipril treatment and the angiotensin II/angiotensin I ratio was reduced at the same time. Our results show that the responses of the plasma and kidney components of the renin angiotensin system to angiotensin-converting enzyme inhibition are different in the plasma and the kidney suggesting that the circulating and tissue renin angiotensin system are at least in part independent. PMID- 9404424 TI - [Insulin resistance and essential hypertension in Vietnamese subjects]. AB - Several epidemiological and experimental studies suggest that essential arterial hypertension is associated with hyperinsulinism and insulin resistance in obese subjects and also in subjects with normal body weight. Undernutrition remains frequent in adult Vietnamese people and mean body mass index is around 18.5 kg/m2 in Vietnam. The aim of this study was to look for insulin resistance in hypertensive Vietnamese subjects, despite a markedly lower BMI in Vietnam than in occidental countries. One hundred and eight hypertensive patients (51 men and 57 women) over 40 years (mean = 65.4 years) were compared with 36 healthy subjects (23 men and 13 women) over 40 years (mean = 63.8 years). Hypertensive patients had significantly higher BMI (20.5 +/- 0.3 (SEM) kg/m2 vs 18.4 +/- 0.4 kg/m2; p < 0.01), thicker triceps skinfold (1.26 +/- 0.07 cm vs 0.71 +/- 0.07 cm; p < 0.001) and not significantly different waist/hip ratio (0.88 +/- 0.01 vs 0.85 +/- 0.01). Blood glucose at fasting and 2 hours after 75 g glucose taken orally were similar in hypertensive and normotensive subjects. Plasma insulin at fasting and 2 hours after glucose were significantly higher in hypertensive patients (44.4 +/- 5.1 pmol/L vs 21.6 +/- 3.2 pmol/L; p < 0.05 and 271.1 +/- 21.6 pmol/L vs 139.1 +/- 15.2 pmol/L; p < 0.001). Thus, despite under-nutrition, hypertensive Vietnamese patients have a moderate but significant increase in BMI and fat mass without predominant abdominal localization, and a state of insulin-resistance, compared with normotensive healthy subjects. PMID- 9404425 TI - [Peripheral insulin resistance and free intralymphocyte magnesium and calcium concentrations in patients with essential hypertension]. AB - To ascertain the claimed links between peripheral insulin resistance and intracellular magnesium and calcium concentrations, we measured free intralymphocyte magnesium (Mg(i)) and calcium (Ca(i)) concentrations, as well as the rate constant of plasma glucose disappearance (K(itt)) after insulin injection (insulin tolerance test: ITT), in a group of 15 normotensive control subjects (NC) and 29 essential hypertensive subjects (EH). Mg(i) and Ca(i) were measured in triplicate by means of a fluorimetric technique based on the dyes furaptra and fura-2 respectively. K(itt) value were significantly reduced in hypertensive subjects as compared to control subjects (M +/- SD, EH: 4.54 +/- 1.31 vs 5.63 +/- 1.07; p < 0.02; 95% confidence limits: 0.22-1.96). Mg(i) and Ca(i) were not statistically different in hypertensive subjects as compared to control subjects (Mg(i), NC: 0.274 +/- 0.02 mmol/L; EH: 0.248 +/- 0.05 mmol/L; Ca(i), NC: 47.6 +/- 9 mmol/L, EH: 46.7 +/- 13.6 mmol/L). A statistically significant inverse correlation was found in the whole study group between K(itt) and body mass index (R = -0.394, p = 0.01) and a statistically significant positive correlation between K(itt) and Mg(i) (R = 0.386; p = 0.012). The latter correlation was no longer statistically significant if adjusted for body mass index. Our data are in favour of a link between index of peripheral insulin resistance and body mass index. A dependence from Mg(i) is possible but the study lack so far the statistical power to demonstrate it. PMID- 9404426 TI - [Autonomic nervous system abnormalities in the initial phase of insulin resistance syndrome. Value of the study of variability of cardiac rate and blood pressure on a model of nutritional obesity]. AB - Changes in the activity of the sympathetic activity are often involved in the development of human insulin-resistance syndrome. However, the nature of changes in both the parasympathetic and orthosympathetic components are still controversial. We have recently developed an experimental model reproducing in dog this morbid triptyque (obesity, hypertension and hyperinsulinism), obtained by hypercaloric hyperlipidic diet. The aim of the present study was to characterize the changes in autonomic nervous system and spontaneous baroreflex in the initial period of obesity-hypertension syndrome. Ten male Beagle-Harrier dogs were used in this study. We investigated before and during 20 weeks after the beginning of the hypercaloric diet, plasma insulin, noradrenaline levels, spontaneous baroreflex efficiency (using the sequence method), arterial blood pressure, heart rate and their spectral analysis (fast Fourier Transformation) in both low (LF: 50-150 mHz, reflecting sympathetic activity) and high (HF: respiratory rate +/- 50 mHz, reflecting parasympathetic activity) frequency bands. Body weight (+20%), systolic (SBP: +23%) and diastolic (+16%) blood pressure and heart rate (+19%) increased during 6 weeks and then remained stable. Concomitantly, high frequency of HR (22.01 +/- 1.9 vs 14.15 +/- 1.04% at 7th week) and BF of systolic blood pressure (15.6 +/- 1.1 vs 19.2 +/- 1.2% at 4th week); p < 0.07, showed a rapid decrease in parasympathetic tone and a early increase in sympathetic activity. Nevertheless, in steady state of this syndrome, parasympathetic tone returned to initial values (18.43 +/- 3.25% at 20th week). Insulinemia significantly increased from the 4th week (14.2 +/- 0.9 vs 25.3 +/- 2.2 microUI/mL at 20th week), but noradrenaline remained not modify (400 +/- 85 vs 312 +/- 45 pg/mL at 20th week). Spontaneous baroreflex efficiency also decreased from the 2nd week (35.5 +/- 5.5 vs 16.7 +/- 4.9 mmHg/ms at 20th week). This study shows that an hyperlipidic hypercaloric diet induces a decrease in both parasympathetic tone and spontaneous baroreflex efficiency, which could be the physiopathological link between obesity, hypertension and hyperinsulinism. PMID- 9404427 TI - [Alteration of cardiovascular vagosympathetic control evaluated by spectral analysis of variations of heart rate and blood pressure in obesity]. AB - Several studies suggest alterations of parasympathetic and sympathetic control in obesity. We have already shown that more than 40% of non diabetic obese subjects have alterations of parasympathetic control of heart rate (HR) variations. The present study aimed to investigate parasympathetic and sympathetic cardiovascular control by using spectral analysis. Sixty-two non diabetic obese subjects were compared to 38 sex-matched healthy controls. Spectral analysis was performed by Anapres system and identified two particular peaks: the one of high frequency (0.20-0.25 Hz) for heart rate variations during controlled breathing which depends on parasympathetic activity, the other of low frequency (around 0.10 Hz) for systolic BP variations in the standing position which mainly depend on sympathetic activity. In control subjects the amplitude of the high frequency peak (r = -0.556, p < 0.0001) but not the amplitude of the low frequency peak correlated negatively with age. In the obese subjects both the high and low frequency peaks correlated negatively with age (r = -0.249; p = 0.05 and r = 0.289, p = 0.036 respectively) and did not correlate with body mass index. The high frequency peak was significantly lower than in controls (4.80 +/- 3.37 (SD) vs 8.38 +/- 4.14; p < 0.0001). In the 25 obese subjects over 40 years the low frequency peak was also significantly lower than in controls (10.00 +/- 3.10 vs 11.95 +/- 4.25; p < 0.05). This study suggests that 1) age must be taken into account when interpreting the cardiovascular parameters under vagosympathetic control; 2) in non diabetic obese subjects vagal activity is decreased and in those over 40 years sympathetic activity is also decreased. PMID- 9404428 TI - [The Hypertension Cochrane Review Group. Presentation and user's guide]. AB - Cochrane collaboration has been developing since 1992 as an international network aiming at performing systematic reviews of available data on therapeutic effectiveness. The fundamental principles of this organisation are trying to avoid duplication of efforts, seeking the best reliability, using reproducible and quantitative synthesis techniques, offering constantly updated results. All health domains are progressively covered. The production unit in one domain is the review group. The Hypertension Cochrane Review Group (HTN CRG) has been officially registered on May 15th 1996. Information and products from the group are available through its news letter, through the Cochrane Library CD-ROM, regularly updated, and on the Internet (http://merece.uthscsa.edu/htncrg). The Hypertension Cochrane Review Group includes an editorial board (with an administrator and three editors), the authors of systematic reviews, internal and external reviewers. The geographic link is the San Antonio Cochrane Centre (Texas, USA). Invitations to participate have been sent to people interested in hypertension and who where known to the Cochrane collaboration, and to authors of previous reviews in hypertension. It is possible to collaborate with the HTN CRG through: performing a systematic review; reviewing protocols and systematic reviews; hand-searching medical journals; being a member of the editorial team. The first protocol for a systematic review edited by the group concerns antihypertensive treatment in the elderly, and is available in the 1996 and subsequent editions of the Cochrane Library. The group welcomes other reviews from domains awaiting registration, and collaborates with related domains review groups such as Diabetes CRG, or Stroke CRG. The group contributes to the effort of hand-searching medical literature, Pr Plouin being responsible for the Archives des Maladies du Coeur et des Vaisseaux. The second edition in 1996 of the Cochrane Library included 114 systematic reviews and 131 protocols, being the only media with similar objectives. PMID- 9404429 TI - [Long-term clinical tolerance of antihypertensive treatment during the HOT study. Groupe francais de l'etude HOT]. AB - The objective of the HOT study, an international, prospective, randomised study is to determine the optimal level of the blood pressure under treatment, in linked with the lowest cardiovascular mortality and morbidity. The target diastolic blood pressure of 80, 85 and 90 mmHg was determined at the randomisation. In order to reach the target blood pressure, a strategy of treatment was determined: the 1st step was felodipine (a long acting dihydropyridine) and the next steps (if the blood pressure reduction is not enough) propose the addition of different therapeutic classes and/or the increase of each drug doses. The available data after 2 years of the patients follow-up allow us to evaluate the incidence of the reported side effects according to the target blood pressure assigned by randomisation and the number of hypertension drugs used to reach these targets. The percentage of patients with at least one side effect at 12 and 24 months of follow-up are respectively: for the target group DBP < or = 90 mmHg: 9.2% versus 6%; for the target group DBP < or = 85 mmHg: 8% versus 4.4%; for the target group DBP < or = 80 mmHg: 7.9% versus 4.9%. The overall tolerability is not influenced by the target diastolic blood pressure but depends on the number of hypertension drugs used. At 24 months, 2.8% of patients are under monotherapy; 7% under bitherapy and 9.8% under tritherapy. The incidence of the side effects decreases after the 1st year, but slower than between the third months and the first year. There is an influence of the region on the incidence of the side effects, the south European countries describing more side effects than France or the north European countries. This seems to be linked with a perception of the side effects more than with a higher rate. In conclusion, these results confirm the possibility to reach a targeted blood pressure using a predetermined strategy without increasing dramatically the incidence of the side effects. PMID- 9404430 TI - [Isolated systolic hypertension and cognitive function in the aged. Experience of the Syst-Eur study]. AB - OBJECTIVE: To determinate cognitive status and its correlates in older patients with isolated systolic hypertension. METHODS: Syst-Eur is a double-blind placebo controlled outcome trial conducted in European patients over 60 years of age with isolated systolic hypertension. Moreover, a side-project--the Vascular Dementia Project--is designed to assess cognitive functions and to follow-up their evolution to determine the influence of antihypertensive therapy on vascular dementia incidence. Cognitive functions were evaluated at entry with the MiniMental State Examination (MMSE) in 2250 patients included in Syst-Eur. Cognitive impairment was defined with a MMSE score < or = 23 and led to further evaluation. Baseline blood pressure (BP) was based on the average of six sitting blood pressure readings at three run-in visits 1 month apart. Statistical analysis used Spearman correlation. RESULTS: The MMSE was analysed in 1374 women and 751 men whose mean age was 70 years (range: 60-100). The median level of education expressed as the age at which they stopped their education at school, was 15 years. Baseline blood pressure averaged 173 +/- 10/86 +/- 6 mmHg. Before randomisation in the trial, 899 (40%) patients had received antihypertensive therapy and 602 (27%) had experienced cardiovascular complications. The MMSE scores ranged from 15 to 30 (median = 29). The maximal score of 30 was reached by 609 (30%) subjects. Among the 59 (3%) patients with a MMSE-score of 23 or less, 5 were considered to be demented according to the DSM IIIR criteria. The MMSE scores decreased with advancing age in men (r = -0.16; p < 0.001) and women alike (r = -0.24; p < 0.001). In both men and women, they were positively correlated with the level of education (r = 0.30 and 0.32, respectively; p < 0.001). They were negatively correlated with systolic blood pressure (r = 0.10; p < 0.001) and slightly positively correlated with diastolic blood pressure (r = 0.05; p = 0.03). Previously treated patients or patients reporting cardiovascular complications at baseline had lower MMSE-scores than their non-treated counterparts or subjects without cardiovascular complications (median = 28 and 29, respectively, p < 0.003). CONCLUSION: In a European cohort of 2225 patients over 60 years of age with isolated systolic hypertension, the level of cognitive functions evaluated with the MMSE decreases with advancing age or lesser educational level. It also decreases with higher systolic blood pressure or lower diastolic blood pressure. The influence of antihypertensive therapy on cognitive status will be prospectively evaluated in Syst-Eur Vascular Dementia Project. PMID- 9404431 TI - [HOT study: quality of the blood pressure control after 2 years follow-up. Pour le Groupe francais de l'etude HOT]. AB - The objective of the HOT study, an international, prospective, randomised study was to determine the optimal level of the blood pressure under treatment, linked with the lowest cardiovascular mortality and morbidity. The target diastolic blood pressure of 80, 85 and 90 mmHg was determined at the randomisation. In order to reach the target blood pressure, a strategy of treatment was predefined: the 1st step was felodipine (a long acting dihydropyridine) and the next steps (if the blood pressure reduction was not enough) proposed the addition of different therapeutic classes and/or the increase of the doses of each drug. The blood pressure measurements were made, using the oscillometric method (automatic blood pressure measuring device, Hestia). The quality of the blood pressure control observed in the HOT study was verified after 6 months of follow-up ("Quality of the blood pressure control in the clinical practice and in the HOT study", for the French research group of the HOT study. French hypertension meeting, Paris, December 1994). The aim of this second evaluation was to see if the quality of this control was still effective in France and for all countries after 2 years of follow-up. At the inclusion, the mean diastolic blood pressure was 106 +/- 4 mmHg in France (n = 1.574) and 105 +/- 4 mmHg for all countries (n = 18.790). The results at 24 months were the following, according to the target groups: 79.9 for the < or = 80 mmHg target group; 82.1 for the < or = 85 mmHg target and 83.6 for the < or = 90 mmHg target group. The percentages of patients who reached the target blood pressure were respectively 74; 80; 89% for the 3 target groups. The number of antihypertensive treatments needed to reach this blood pressure control slightly increased in the 3 target groups between the first and the second year with a lower rate of monotherapy and a higher rate of bi and tritherapy. But in the 80 mmHg target group (the most strict), the monotherapy was used in more than half of the patients. In comparison with all countries, France had lower number of bi and tritherapies (i.e. in the 85 mmHg target group: 38.4% of bitherapy in France versus 45.6% in all countries). CONCLUSION: after 2 years of follow-up, the quality of the blood pressure control is still good. There is a trend toward a slight increase in the number of antihypertensive drugs after the first year in the 3 target groups. PMID- 9404432 TI - [Hypertension and primary glomerulonephritis in adults. A study of 302 cases]. AB - The purpose of the present work was to show the place of hypertension in primary glomerulonephritis in adults. Hypertension was defined as diastolic blood pressure above 90 mmHg and renal insufficiency as serum creatinine above 135 mc mol/L. Secondary glomerulonephritis was excluded. The study was performed in 302 patients with primary glomerulonephritis biopsied between March 1994 and March 1996. They were 183 males and 119 females, aged from 16 to 63 years (mean: 29.8 years). The incidence of hypertension at the time of admission was 46.6%: 141/302 cases. The only consideration of prolonged hypertension (excluded transient hypertension of acute nephritic syndrome) shows an incidence of 31.4%: 95/302 cases (table). Frequency of hypertension (HT) in different types of primary glomerulonephritis (GN): [table: see text] The histological types observed in these cases of hypertension were represented essentially by the proliferative lesions: 73% (72/95 cases) who were grouped mainly in proliferative glomerulonephritis postinfectious and IgA nephropathy. No proliferative lesions: 24% (23/95 cases) were especially represented by focal segmental sclerosis. Renal insufficiency noted in 69 cases on 95 hypertensions was probably the result of the parallel evolution of hypertension renal lesions and those belonging to these histologic types. In conclusion, this study shows a narrow correlation between the hypertension and proliferative glomerulonephritis in our young adults population. PMID- 9404433 TI - [Para-aortic paraganglioma found by arteriography. Apropos of a case and review of the literature]. AB - A 53 years old man had an angiography for suspected renovascular hypertension (arteritis, renal insufficiency, duplex scanning). It showed a narrow right renal artery streched by a 45 mm mass arising from the adrenal. The computed tomography showed the tumor and the nuclear magnetic resonance imaging indicated a pheochromocytoma. The patient had no complain of headaches, palpitations or sweating. Biochemistry was normal except for a slight serum creatinin elevation and a non significant urinary noradrenaline level. A diagnostic of non functioning pheochromocytoma was made. The therapeutics consisted in a surgical ablation of the tumor and the right kidney (non functioning) and the patient became normotensive thereafter without treatment. The histologic feature was an aortico-sympathetic paraganglia, the adrenal was normal. Paraganglias are arising from the paraganglion system including chemodectoma and glomus jugulare tumor. Non functioning retroperitoneal paraganglias are uncommon: less than 50 in the literature between 1902 and 1992. PMID- 9404434 TI - [Evaluation of the quality of filling out medical records in a hypertension consultation]. AB - The aim of the study was to evaluate the completion of medical records of a hypertension clinic and to compare standardized computerized records versus standard medical records. The medical records of 163 consecutive hypertensive patients attending at the Broussais hospital hypertension clinic between December 1995, 6th and January 1996, 21st were checked. At the last visit, the patients were attended by 16 physicians working in 4 different teams. The medical data were recorded by physicians in the computerized system called ARTEMIS in 120 patients and in standard structured forms in 43 patients. The patients notes were checked to see if 9 clinical items were recorded at the first visit (V1), at the visit before last (V2) and at the last visit (V3). The overall completion rate was high at V1 (92.2%) and significantly decreased at follow-up visits (82.6% at V2 and 83.2% at V3). The completion rate was significantly higher in the computerized records than in the standard notes: 95.8% vs 82.2% at V1, 91.9% vs 56.3% at V2 and 91.6% vs 59.7% at V3. During follow-up (V2 vs V1), a significant decrease in the completion rate of 6 items was observed in the standard notes (tobacco use, alcohol consumption, physical activity, compliance to treatment, body weight, manual blood pressure measurement). In the computerized records, only physical activity completion rate decreased. In conclusion, the computer may help to increase the quality of the medical records as reflected by the completion rate of items related to hypertension care. PMID- 9404435 TI - Gonadal function in men with chronic illness. PMID- 9404436 TI - Hypothalamic adipsic syndrome: diagnosis and management. AB - Patients with hypothalamic adipsic syndrome, especially in conjunction with diabetes insipidus, pose management difficulties. They are at risk of both under- and over-hydration. We present 4 patients with hypothalamic adipsic syndromes, due to different causes, illustrating the practical difficulties encountered in this condition. The principles of management, with a sliding scale of water intake related to changes in daily body weight, are discussed. PMID- 9404437 TI - Thyroid function in very low birth weight infants. AB - OBJECTIVES: The purpose of this study was to test the hypothesis that low circulating thyroxine concentrations characteristic of very low birth weight (VLBW) neonates (< 1500 g) are the result of decreased protein binding of thyroid hormones and to elucidate the mechanism(s) responsible and possible significance thereof. DESIGN: Cross-sectional comparison of thyroid related measurements in cord blood specimens from VLBW infants and from full term infants. Longitudinal comparison in cord and 2- and 4-week blood specimens from VLBW infants. PATIENTS: Cord blood specimens were analysed from 47 VLBW and 45 full term infants weighing > or = 2500 g. Repeat analyses in venous bloods from 32 of the VLBW infants were analysed at 2 weeks of age and again at 4 weeks in 23. The first cohort of patients was studied in 1994 and comprised 28 VLBW and 24 full term infants (Cohort A). The studies were repeated in 1995-96 in 19 VLBW infants and 21 full term infants (Cohort B). MEASUREMENTS: T4, free T4 (FT4), T3, thyroxine binding globulin (TBG), and TSH were measured in cord blood and 2- and 4-week venous specimens from VLBW infants and in cord blood specimens of full term infants. Molar ratios of T4/TBG were calculated. RESULTS: (1) Cord blood TBG, T4 and T3 concentrations of VLBW infants were each 60% of those of term infants. TBG concentrations were 397 +/- 111 vs 680 +/- 172 nmol/l (P < 0.0005). T4 concentrations were 76 +/- 22 vs 139 +/- 26 nmol/l (P < 0.0005). FT4 concentrations were in the normal adult range in both neonatal groups. T4/TBG ratios did not differ between the neonatal groups but were significantly less than that of adults (P < 0.001). (2) TSH concentrations in VLBW infants at 2 and 4 weeks were less than 50% of cord blood values. At 2 weeks, TBG concentrations of VLBW infants were unchanged from cord blood concentrations but mean T4 concentration fell by 18% and T4/TBG ratios by 21% (P < 0.005). Mean FT4 rose by 78% (P < 0.02). The changes in mean T4 and FT4 were due largely to FT4 concentrations of 37-113 pmol/l and T4 concentrations of 13-48 nmol/l in 5 infants. These infants also had lower T4/TBG ratios and were smaller and more ill than the remainder of the cohort. The changes disappeared by 4 weeks in 3 of the 4 infants tested. CONCLUSIONS: Cord T4/TBG ratios are the same in very low birth weight and term infants and are significantly lower than in adult blood. These are more than compensated for in term infants by a 236% increase in thyroxine binding globulin concentrations. The lower thyroxine binding globulin concentrations in very low birth weight infants explain their much lower T4 concentrations. Cord FT4 concentrations of full term and very low birth weight infants are in the normal adult range. T4 concentrations are further depressed and free T4 concentrations elevated in the most ill very low birth weight infants at 2 weeks of age in a manner analogous to that of the 'sick euthyroid syndrome'. PMID- 9404438 TI - Thyroid function in very low birthweight infants. PMID- 9404439 TI - Is polycystic ovary syndrome a neuroendocrine or an ovarian disorder? PMID- 9404440 TI - The effect of propylthiouracil on subsequent radioactive iodine therapy in Graves' disease. AB - OBJECTIVE: Antithyroidal drugs (ATD) are used in the management of Graves' disease either as primary therapy for several months while awaiting remission of the disease or as pretreatment for several weeks prior to definitive radioactive iodine therapy (RAI). We have reported previously that pretreatment with propylthiouracil (PTU) before definitive RAI therapy is associated with a higher RAI treatment failure rate than RAI therapy alone. The objectives of the current study were 2-fold. First, to verify the results of our prior study regarding the effect of PTU used as pretreatment before RAI in a cohort of patients from a different institution and, secondly, to better define the relationship between the number of days off PTU before RAI therapy and therapeutic efficacy of RAI dosing. DESIGN: A retrospective review of Graves' disease patients treated from 1980 to 1994. PATIENTS: Study patients had to meet the following inclusion criteria: radionuclide studies and thyroid hormone values consistent with Graves' disease, at least 1 year of follow-up data available and discontinuation of the ATD at least 4 days before RAI administration. Exclusion criteria included therapy with any ATD other than PTU or ATD therapy during or following RAI dosing. MEASUREMENTS: Effectiveness of RAI therapy, days on PTU, days off PTU and calculated RAI dose to the thyroid were recorded for each subject. We compared the efficacy of RAI therapy in patients treated with PTU (used either as pretreatment in preparation for RAI therapy or as primary long-term therapy) before RAI administrations to those treated with RAI alone with special attention to the number of days on and off PTU before RAI dosing. Patients were considered RAI treatment failures if a second dose of RAI was required to achieve a euthyroid or hypothyroid state. RESULTS: One hundred and sixteen patients met our study criteria. Forty patients received PTU therapy for a mean of 221 +/- 59 days. The PTU was discontinued for a mean of 60 +/- 25 days before RAI dosing. Persistent hyperthyroidism was seen in 9% (7/76) of patients treated with RAI alone. The failure rate of a single dose of radioactive iodine was significantly increased when PTU was discontinued between 4 and 7 days before the administration of RAI (29% vs 9% for RAI alone, P = 0.039). PTU discontinued for at least 1 week before RAI dosing was associated with a nearly 2-fold increase in failure rate, but this difference did not achieve significance (17% vs 9% for RAI alone, P = 0.24). Examining only those patients receiving PTU, patients who had successful single dose RAI therapy tended to receive a higher dose of RAI than patients failing RAI therapy (480 +/- 30 vs 410 +/- 40 MBq administered dose, P = 0.18; and 8.0 +/- 0.9 vs 5.5 +/- 1.1 MBq/g thyroid tissue calculated dose, P = 0.21). Furthermore, total serum thyroxine at diagnosis was significantly higher in patients failing RAI therapy after PTU administration than in patients successfully treated with RAI after receiving PTU (316 +/- 40 vs 225 +/- 13 nmol/L, P = 0.03). CONCLUSIONS: Propylthiouracil discontinued 4-7 days before radioiodine dosing is associated with a significant increase in the failure rate of a single dose of radioiodine. Discontinuation of the propylthiouracil for at least a week before radioiodine administration is associated with a higher, although not statistically significant, radioiodine failure rate. In patients that require treatment with propylthiouracil before radioiodine therapy, a higher total serum thyroxine level at diagnosis is associated with an increased rate of radioiodine failure. Consideration should be given to increasing empirically the dose of radioiodine administered to Graves' disease patients that have received propylthiouracil within a week of radioiodine administration in an effort to decrease the radioiodine failure rate to an acceptable level. PMID- 9404442 TI - Quality of life, body composition and muscle strength in adult growth hormone deficiency: the influence of growth hormone replacement therapy for up to 3 years. AB - OBJECTIVE: Adults with GH deficiency complain frequently of low energy levels, emotional lability and mental fatigue resulting in a low perceived quality of life (QOL). Body composition is altered with increased fat mass and decreased lean body mass and muscle strength is reduced. The aims of this study were to determine the effects of replacement GH treatment on: (a) body composition and muscle strength and (b) QOL, using specifically selected and adapted measures. DESIGN: A 12-month study (double-blind placebo-controlled for the first 6 months and open for the second 6 months) of GH replacement injections (0.125 iu/kg/week for the first month and 0.25 iu/kg/week for the following 5 months of each study period) in GH deficient adults on QOL, body composition and muscle strength. This was followed by an open study of a further 12 months' GH treatment assessing QOL and muscle strength. Finally, QOL was assessed after up to 3 years of GH replacement treatment. PATIENTS: Thirty of the 32 adult patients with GH deficiency enrolled completed the initial 12-month study (10 male, mean age 33.5 years, mean (SD) stimulated serum GH response 3.0 mU/l (2.86)). Nineteen patients then opted to continue GH treatment. Of these, 13 patients were available for assessment after a further 12 months' and 24 months' treatment. MEASUREMENTS: Health-related QOL was assessed using 2 specifically adapted scales for adults with GH deficiency: the Life Fulfillment Scale and the Impact Scale. In addition 4 other self-rating questionnaires were used: Nottingham Health Profile, Hospital Anxiety and Depression Scale, Self Esteem Scale and Mental Fatigue Scale. Body composition was assessed by DEXA and quadriceps muscle strength by measuring maximum voluntary contractions. RESULTS: In the initial 12 months' placebo controlled study perceived energy levels increased after 6 and 12 months of GH treatment (P < 0.01 compared with baseline) in the patients receiving GH for the full 12-month period. There were no changes in energy levels throughout the study in the group receiving placebo for the first 6 months. Also small improvements in impact scores were found after 6 months of GH treatment (P < 0.05) but this was not sustained at 12 months. In both GH and placebo groups life fulfillment worsened after 6 months, but then improved to baseline values after 12 months. In the patients who persisted with GH replacement, energy levels continued to improve (at 2 years, P < 0.01 compared with baseline) but then fell (at 3 years, P = NS compared with baseline). A similar pattern was observed in emotional reaction scores. However, improvements in self-esteem were maintained (at 3 years, P < 0.05 compared with baseline). Body composition altered favourably over the initial 12-month study period with a significant increase in lean mass and decrease in fat mass in both groups after 6-12 months of GH. There were no changes in muscle strength in either group during the initial 12-month study. However, in the patients who were available for assessment after a further 12 months of GH treatment, muscle strength increased significantly (P < 0.02 compared with baseline). CONCLUSION: GH replacement treatment for 6-12 months leads to significant improvements in body composition (DEXA) but longer-term treatment may be needed to increase muscle strength. Self-esteem scores improve and are maintained after 3 years of treatment. Energy levels and emotional reaction improve during treatment for up to 2 years but decline thereafter. PMID- 9404441 TI - Pituitary dysfunction in recently post-menopausal women. Nottingham EPIC Study Group. AB - OBJECTIVE: Some recently post-menopausal women have lower than expected FSH concentrations, raising the possibility of subclinical hypothalamo-pituitary impairment. We have therefore performed pituitary stimulation tests in a group of recently post-menopausal women recruited to a bone loss prevention study. DESIGN: Prospective study of health volunteers. SETTING: Outpatient, teaching hospital in Nottingham UK. SUBJECTS: Forty-seven women selected from a cohort of 428 healthy volunteers to an osteoporosis prevention study all within 10 years of the menopause. MAIN OUTCOME MEASURES: Response of the pituitary to formal stimulation tests and its relationship to bone mineral density. RESULTS: A significantly attenuated response to pituitary stimulation was found in a group of otherwise healthy women with an FSH considered to be inappropriately low for the level of circulating oestradiol. The impaired responses were significant not only for FSH as expected but also to LH and prolactin at 1 hour after injection of GnRH and TRH (area under the curve, FSH P = 0.01, LH P = 0.001, prolactin P < 0.0001). TSH secretion was not significantly impaired. Baseline cortisol, growth hormone and thyroxine (T4) were normal in both control and test subjects. The test group was both heavier and taller, with a higher lean body mass, a higher body mass index and a greater total body fat than the controls. Bone mineral density showed no significant differences between the groups. Test subjects also had a higher free oestradiol index (P < 0.001) which correlated strongly (r = 0.534, P = 0.00026) with baseline FSH levels and possibly reflects a greater tissue exposure to biologically active hormone. Oestrone concentrations were, however, no different between the groups. CONCLUSIONS: Serum FSH concentrations are not invariably elevated in recently post-menopausal women and use of FSH as a determinant for postmenopausal status in clinical trials should be used with caution. Notwithstanding their higher free oestradiol index, women found to have an abnormally low basal FSH had evidence of poor prolactin, FSH and LH but not TSH responses to pituitary stimulation. This may represent either a degree of subclinical pituitary failure of a variant of normal. The low levels of gonadotrophin activity did not affect bone mineral density. PMID- 9404443 TI - Urinary growth hormone: a screening test for growth hormone sufficiency. AB - OBJECTIVES: The majority of short statured children referred for serum GH testing prove to be GH sufficient. The purpose of our study was to evaluate urinary growth hormone (uGH) as a screening test for GH sufficiency. PATIENTS: We studied (i) short statured children previously diagnosed as GH sufficient (n = 44) or GH deficient (n = 41) (peak serum GH > or = 8 micrograms/l or < 8 micrograms/l, respectively); (ii) short children undergoing serum GH stimulation tests (n = 23, test group); (iii) normal statured children (n = 45, control group). DESIGN: Three separate overnight urine collections were obtained in all groups. GH injections in GH deficient subjects were discontinued 4 days prior to urine collection. MEASUREMENTS: uGH concentrations were measured using a chemiluminescence immunoassay. Overnight uGH was expressed in several ways (overnight excretion and overnight excretion corrected for body surface area, time and creatinine). Receiver operator curves (ROC) were constructed from the data obtained in the GH sufficient and deficient subjects. Sensitivity and specificity were then determined for various urinary cut-offs. These cutoffs were validated in turn in the test group by comparison of the predicted with the observed GH status. RESULTS: The GH deficient group had the lowest GH output with respect to overnight uGH, overnight uGH/m2, overnight uGH/h and overnight uGH/creatinine when compared with the GH sufficient and control groups (P = 0.0001). Overnight uGH/m2 data gave the greatest area under the ROC curve. At 100% specificity (no GH deficient subjects), it had the highest sensitivity, 63.6% (49.2-78.0% CI) at a cut-off of 2.3 ng/m2 (63.6% of GH sufficient subjects had uGH levels > 2.3 ng/m2). When this and other cut-offs were applied to the test group, we found consistency between the observed and predicted numbers of GH sufficient and deficient subjects. CONCLUSIONS: We conclude that urinary GH is a useful test for the diagnosis of GH sufficiency as defined by serum criteria and can be used to reduce significantly the number of serum stimulation tests. PMID- 9404444 TI - Anti-GAD65 autoantibody in Taiwanese patients with insulin-dependent diabetes mellitus: effect of HLA on anti-GAD65 positivity and clinical characteristics. AB - OBJECTIVE: Anti-GAD65 antibody has been studied widely in patients with insulin dependent diabetes mellitus (IDDM) in many different populations. However, the prevalence of GAD65 autoantibody has not been assessed in Taiwanese patients with IDDM. We therefore characterized GAD65 antibody and investigated the effect of HLA-DR phenotypes on GAD65 autoimmunity and other clinical characteristics in Taiwanese subjects with IDDM. SUBJECTS AND MEASUREMENTS: Two hundred and twenty five patients (male 102, female 123) with IDDM were recruited. The diagnostic criteria for IDDM were age of onset before 30 years, presence of diabetic ketoacidosis, and insulin-dependency within 3 years of onset. We employed a radioligand method to detect GAD65 antibody. HLA-DR typing was performed by the PCR-SSO techniques. Plasma C-peptide and anti-thyroid microsomal antibody were also measured. RESULTS: The prevalence of GAD65 antibody according to duration of disease were 50/91 (54.9%), 37/95 (38.9%), 8/24 (33%), and 3/15 (20%) among the groups of duration < or = 5, 6-10, 11-15, and > 15 years, respectively (p = 0.0011). There were no significant differences between GAD(+) and GAD(-) patients in age of onset (11.5 +/- 6.5 and 11.6 +/- 13.4 years, respectively), gender distribution (male:female 39:59 and 58:69, respectively) and percentage with residual beta cell function (38.8% and 29.1%, respectively). Multiple regression analysis revealed that duration of IDDM correlated inversely with residual beta cell function. Earlier onset of IDDM correlated with a loss of beta cell function and a HLA-DR phenotype containing DR3/4, DR3/3 or DR3/9. CONCLUSIONS: Prevalence of GAD65 autoantibody among Taiwanese subjects with IDDM was negatively correlated with duration of disease. Different determinants in the HLA-DR locus contributed to the clinical onset of IDDM but not to GAD autoimmunity. PMID- 9404445 TI - Abnormal melatonin secretion in hypogonadal men: the effect of testosterone treatment. AB - OBJECTIVE: We have recently demonstrated that GnRH deficient male patients have increased nocturnal melatonin secretion, whereas hypergonadotrophic hypogonadal males have decreased melatonin levels. We were interested in determining whether testosterone (T) treatment (when T levels were well matched with pubertal control values) has an effect on melatonin secretory profiles in these patients. DESIGN: Prospective, controlled. SUBJECTS: Six male patients with idiopathic hypogonadotrophic hypogonadism (IGD), six males with hypergonadotrophic hypogonadism due to Klinefelter's syndrome (KS) and seven controls. Patients were examined before and during the administration of 250 mg testosterone enanthate/month for four months. MEASUREMENTS: Serum samples for melatonin levels were obtained every 15 minutes from 1990 to 0700 h in a controlled light-dark environment. The results of FSH, LH, T and oestradiol (E2) (determined at hourly intervals) and melatonin profiles, were compared with the pre-treatment values in each group, and with values obtained in the control group. RESULTS: All 12 patients had low pre-treatment T levels (1.4 +/- 0.7 in IGD and 2.0 +/- 0.4 in KS vs. 19.8 +/- 2.3 nmol/l in controls) and attained normal levels after four months of T treatment (19.5 +/- 7 in IGD and 22.7 +/- 3.8 nmol/l in KS). Serum LH, FSH and E2 levels (11 +/- 4 IU/l, 24 +/- 10 IU/l and 113 +/- 12 pmol/l, respectively) were still elevated in KS during T treatment as compared with values in controls (2 +/- 1 IU/l, 2 +/- 1 IU/l and 67 +/- 4 pmol/l, respectively). In IGD, serum LH (0.12 +/- 0.1 IU/l) and FSH (0.16 +/- 0.2 IU/l) levels during T treatment were suppressed. Pretreatment melatonin levels in IGD were greater than those in age matched pubertal controls while in KS, melatonin levels were lower than values in controls. Melatonin levels were equal in all 12 hypogonadal patients and controls when T levels were well matched. Mean (+/- SD) dark-time melatonin levels decreased from 286 +/- 18 to 157 +/- 26 pmol/l in IGD and increased from 92 +/- 19 to 183 +/- 48 pmol/l in KS (vs 178 +/- 59 pmol/l in controls). The integrated melatonin values decreased in IGD (from 184 +/- 14 to 102 +/- 21 pmol/min. 1 x 10(3)) and increased in KS (from 64 +/- 13 to 123 +/- 40, vs. 116 +/- 39 pmol/min. 1 x 10(3) in controls). No correlations were found between melatonin and LH, FSH or E2 levels. CONCLUSIONS: These data indicate that male patients with GnRH deficiency have increased nocturnal melatonin secretion while in hypergonadotrophic hypogonadal males melatonin secretion is decreased. Testosterone treatment normalized melatonin concentrations in these patients. Taken together, the results suggest that GnRH, gonadotrophins and gonadal steroids modulate pineal melatonin in humans. PMID- 9404446 TI - Abnormal pulsatile secretion of growth hormone in non-insulin-dependent diabetes mellitus. AB - OBJECTIVE: Studies of GH secretion in patients with non-insulin dependent diabetes mellitus (NIDDM) have produced conflicting results. We aimed to differentiate the effects of obesity and metabolic control on pulsatile GH secretion in patients with NIDDM. DESIGN: Blood sampling every 15 min from 22.00 hours to 08.00 hours after a fasting period of at least 3 h. PATIENTS: 13 male NIDDM patients, 9 healthy control subjects matched for age and BMI, and 6 lean subjects matched for age. MEASUREMENTS: Measurement of GH by a novel ultrasensitive chemiluminescence assay. Analysis of concentration vs time profiles by a multiparameter deconvolution technique. RESULTS: GH burst frequency was increased in the NIDDM (0.82 +/- 0.28 h-1) compared with both control groups (lean: 0.6 +/- 0.11; obese: 0.56 +/- 0.19). GH burst mass was decreased in patients (1.57 +/- 0.98 micrograms/l.min) and in obese controls (1.46 +/- 1.44) compared to lean controls (3.71 +/- 3.88). These differences resulted in a significantly higher nocturnal pulsatile GH secretion rate in the lean compared to the obese controls, whereas in the patient group enhanced GH burst frequency compensated for reduced burst mass. The characteristics of GH secretion were not related to nocturnal or early morning blood glucose concentrations. However, GH secretion rate was correlated positively with HbA1c (r = 0.57; P = 0.04), and negatively with plasma C peptide concentrations. CONCLUSIONS: The specific increase in GH burst frequency previously described in insulin-dependent diabetes mellitus is also present in NIDDM. However, GH hypersecretion does not occur because GH burst mass is reduced in proportion to the degree of obesity. The effect of diabetes on the hypothalamic control of GH release appears to be determined by the quality of long-term glycaemic control. PMID- 9404447 TI - Immunoreactive endothelin in nodular pathology of the thyroid. AB - OBJECTIVES: Endothelins (ETs) can act as autocrine and/or paracrine regulators of thyroid homeostasis and growth. The aim of this study was to evaluate immunoreactive ET (i-ET) levels in a group of patients with nodular pathology of the thyroid and to correlate them with the cytomorphological features after fine needle aspiration (FNA) and with hormonal and immunological status and blood pressure levels. DESIGN: Plasma and cystic i-ET were assayed in a group of patients with varying thyroid function, who underwent FNA for solid and cystic nodular pathology. PATIENTS: 47 patients (32-81 years) with nodular pathology of the thyroid and 18 controls (28-70 years) with normal thyroid function and morphology were studied. MEASUREMENTS: Fasting venous blood samples were collected and the plasma for i-ET was frozen at -80 degrees C until assayed. Sera were frozen at -20 degrees C for FT3, FT4, TSH, TPO autoantibodies and thyroglobulin autoantibodies assay. Cystic fluid was obtained by FNA, centrifuged, and the supernatant was stored at -20 degrees C until i-ET assay. FNA cytology was examined by light microscopy. RESULTS: In patients with cystic nodules, plasma i-ET levels were significantly (P = 0.002) higher (5.7 +/- 1.1 ng/l, +/- SEM) than in both patients with solid nodules (2.6 +/- 0.4 ng/l) and (P = 0.02) controls (3.0 +/- 0.3 ng/l). In patients with cystic nodules, cystic i-ET levels (12.6 +/- 1.9 ng/l) were significantly (P = 0.003) higher than plasma levels (5.7 +/- 1.1 ng/l) and did not correlate with the percentage of FNA cellularity. i-ET levels in cystic fluid (12.6 +/- 1.9 ng/l) were significantly (P = 0.0001) higher than plasma i-ET levels in both patients with solid nodules and controls. No difference in either plasma or cystic i-ET levels was found in patients with cystic nodules in relation to differences in thyroid function. No difference in plasma i-ET levels was found between patients with solid nodules and controls. In controls, no significant difference in plasma i-ET levels was found between males and females. A negative correlation (r = -0.55, P = 0.02) was found between cystic i-ET levels and systolic and diastolic blood pressure. No correlation between cystic or plasma i-ET levels and FT3, FT4 or TSH was found in any of the subjects studied. CONCLUSIONS: It seems that endothelins do not possess a primary role in determining thyroid function and that the increased levels in cystic fluid found in our subjects could be secondary to cystic nodule development. PMID- 9404448 TI - Two years of replacement therapy in adults with growth hormone deficiency. AB - OBJECTIVES: Although several studies have shown beneficial short-term effects of recombinant human growth hormone (rhGH) therapy in adult GH deficient (GHD) patients, few data are available on large groups of patients treated for more than one year. In addition, the optimal dose of rhGH for each patient and the baseline parameters that predict which patients will benefit most from therapy or will have adverse events are not entirely elucidated. DESIGN: 148 adult GHD patients were enrolled in a multicentre 2-year rhGH replacement study which was placebo controlled for the first six months. rhGH (Genotropin/Genotonorm Pharmacia & Upjohn) was given in a dose of 0.25 IU/kg/week sc (1.5 IU/m2/day). MEASUREMENTS: Every 3-6 months body composition was measured using body impedance analysis and general well being was assessed using the Nottingham Health Profile (NHP) and social self-reporting questionnaire. At the same time patients had a full clinical examination and blood was sampled for glucose, HbA1c, IGF-1, creatinine, full blood count, thyroid hormones and liver function tests. RESULTS: With rhGH therapy IGF-1 levels increased from -2.00 +/- 2.60 SDS to 1.47 +/- 2.6 SDS after six months (P < 0.001), continued to rise despite no change in dose to 1.84 +/- 2.8 SDS after one year and remained constant thereafter (1.98 +/- 2.4 after 2 years). 56% of patients ultimately attained supranormal IGF-1 levels (+2 SD), 22% had levels below the mean, of which 9% were below -2 SD. Within 3 months lean body mass (LBM) increased by +5.09% (P < 0.001), total body water (TBW) by +5.40% (P < 0.001), while body fat (BF) dropped by -10.89% (P < 0.001) and waist circumference by -1.42% (P < 0.004). These effects were maintained during the first year of therapy, but the effect was attenuated after 24 months: LBM, +3.91% (P < 0.001); TBW, +3.28%, P < 0.001, BF, -6.42% (P < 0.001) and waist -2.22% (P < 0.009). Individual differences in response were large and could not be predicted by any of the baseline parameters, except for a better response in males. Treatment resulted in a large and progressive improvement on the NHP scale, especially energy, emotions and sleep, but a similar change was also found in patients during placebo treatment. With rhGH the number of full days of sick leave/6 months decreased from 12.17 +/- 3.90 days (SEM) to 7.15 +/- 3.50 days after six months (P = 0.009), 2.93 +/- 1.55 days after 12 months (P = 0.01), 0.39 +/- 0.17 days after 18 months (P < 0.001) and 3.3 +/- 2.51 days after 24 months (P = 0.026). Similarly, the hospitalization rate went down from 14.9 to 7% after 6 months and remained at this level thereafter (P = 0.12). About one third of patients on rhGH experienced fluid-related adverse events, most often within the first 3 months. They usually disappeared spontaneously or responded well to dose reduction. Cumulative dropout rates were 29% after 1 year and 38% after two years. Two thirds of these patients stopped treatment because of insufficient subjective improvement. Neither drop-outs nor fluid retention could not be predicted by any of the baseline parameters. CONCLUSIONS: We confirmed in a large group of patients the beneficial effects of rhGH therapy on body composition, metabolic parameters and general well-being and found a consistent drop in number of sick days and hospitalization rate. These effects were maintained during two years of therapy, except for an attenuation in body composition changes after 24 months. The high incidence of fluid-related adverse events suggests that it may be better to start with lower doses of rhGH and to increase the dose more slowly over a number of weeks. The finding of suboptimal high or low IGF-1 levels in many patients reinforces guidelines not to give rhGH in a weight-dependent dose but to titrate it individually for each patient. PMID- 9404450 TI - Open letter to scientific journals, the pharmaceutical industry and drugs approval authorities. ESPE Drugs and Therapeutics Committee. European Society of Paediatric Endocrinology. PMID- 9404449 TI - Low hexarelin dose and pyridostigmine have additive effect and potentiate to the same extent the GHRH-induced GH response in man. AB - OBJECTIVES: Hexarelin (HEX) is a synthetic hexapeptide belonging to the growth hormone-releasing peptide (GHRP) family. The exact mechanism underlying the strong GH-releasing activity of GHRPs is still unclear, though it has been shown that they act both at the pituitary and the hypothalamic level, where they have specific receptors. To clarify the influence of the cholinergic system on the GH releasing activity of GHRPs in man, we investigated the effects of pyridostigmine, a cholinergic agonist which stimulates GH secretion by inhibiting somatostatin release, on the GH response to various HEX doses. DESIGN: We studied the GH release induced by various HEX doses (0.25, 0.5 and 2.0 micrograms/kg i.v.) and pyridostigmine (PD, 120 mg po), both alone and coadministered. The interactions between the lowest HEX dose or PD and the maximally effective GHRH dose (1.0 microgram/kg i.v.) were also studied. SUBJECTS: Six normal male volunteers, aged 24-30 years, were studied. MEASUREMENTS: Serum GH was measured in duplicate by immunoradiometric assay. RESULTS: The GH response to HEX administration was dose-dependent. In fact, the GH response to 0.25 microgram/kg HEX (AUC, mean +/- SEM: 816.4 (235.6 mU/l/120 min) was lower, although not significantly, than that to 0.5 microgram/kg HEX (2154.6 +/- 491.6 mU/l/120 min), which, in turn, was lower (p < 0.05) than that after 2.0 micrograms/kg HEX (4819.2 +/- 668.0 mU/l/120 min). The GH rise after GHRH (1299.2 +/- 222.8 mU/l/120 min) was lower (P < 0.05) than that after 2.0 micrograms/kg HEX, but not different from the responses to either 0.25 or 0.5 microgram/kg HEX. PD induced a significant GH rise (559.0 +/- 129.8 mU/l/120 min, P < 0.05 vs saline), similar to that after 0.25 microgram/kg HEX, and lower than those after both 0.5 and 2.0 micrograms/kg HEX (P < 0.05 and p < 0.01, respectively) and GHRH (p < 0.05). PD pretreatment enhanced the GH response to the lowest HEX dose (1961.4 +/- 253.8 mU/l/120 min, p < 0.05) in an additive way, but failed to modify the GH response to either 0.5 or 2.0 micrograms/kg HEX (2753.6 +/- 444.6 and 5179.0 +/- 770.8 mU/l/120 min, respectively). Notably, the GH response to 0.25 microgram/kg HEX + PD was still lower (P < 0.05) than that to 2.0 micrograms/kg HEX. PD pretreatment as well as 0.25 microgram/kg HEX truly potentiated the GH response to GHRH to the same extent (4926.6 +/- 912.8 mU/l/120 min, p < 0.05 and 5958.8 +/- 750.0 mU/l/120 min, p < 0.05 respectively). The GH responses to PD + GHRH and 0.25 microgram/kg HEX + GHRH were similar to that after 2.0 micrograms/kg HEX alone. CONCLUSIONS: Our results demonstrate that pyridostigmine is able to enhance the GH response only to a very low dose Hexarelin which, in turn, potentiates the GHRH-induced GH rise to the same extent as pyridostigmine. As there is evidence that GHRPs do not inhibit hypothalamic somatostatin release, these findings are consistent with the hypothesis that they act by antagonizing somatostatin activity and/or through unknown factors. On the other hand, though there is evidence showing that GHRH activity is needed for GHRP action, our findings indicate that GHRPs act, at least partially, independently of GHRH. PMID- 9404451 TI - Cushing's syndrome secondary to ectopic ACTH secretion from a primary ovarian carcinoma. PMID- 9404453 TI - [Isotopic signatures in ancient bone collagen]. AB - Collagen can be preserved during tens of thousands of years in bones and teeth under favorable conditions. Natural isotopic abundances in carbon (13C/12C) and in nitrogen (15N/14N) of ancient bone and tooth collagen correspond to those recorded during the biogenic synthesis and have not been significantly altered during fossilization. These isotopic abundances are linked to those of the proteic fraction of animal and human diets, and to physiological conditions. Three kinds of applications are made possible through these natural isotopic signatures: determination of subsistence strategies in ancient human populations, determination of the diet of extinct species and the analysis of past environmental changes. PMID- 9404454 TI - [Molecular archaeology: familial relationship into a neolithic deposit]. AB - At the neolithic period, collective burial-sites were common. In these deposits, we can observe a breakdown of bodies according to age and sex. The nature of this selection is completely unknown. It is a cultural or a family-based selection? We decided to approach this problem by doing a genetic study of the skeletons of the collective deposit of Conde-sur-Ifs (7,000 years B.P., Calvados, France). The study based on the variability of the mitochondrial DNA control region shows that the five individuals studied have all different mitochondrial types. Thus, a familial relationship between these individuals (on a maternal lineage) can be excluded. PMID- 9404452 TI - Growth hormone and serum leptin in GH-deficient adults. PMID- 9404455 TI - [Helminth eggs: trace elements of neolithic and paleolithic parasitosis on French sites]. AB - On the neolithic site of Chalain (Jura, France), the analysis of human coprolites has revealed the presence of many well preserved eggs of Helminths: eggs of Trichuris spP., Capillaria spp., Fasciola hepatica, Diphyllobothrium spP. In the paleolithic picturial sanctuary of the Grande Grotte at Arcy-sur-Cure (Yonne, France), eggs of Ascaris spP. have been discovered. The presence of these parasits open a new way of search about the knowledge of ancient populations. PMID- 9404456 TI - [History of the rabbit and ancient DNA]. AB - Present populations of Rabbits (Oryctolagus cuniculus) are organized into two well defined groups A and B according to their mitochondrial DNA sequences. Group A is restricted to the South Western part of the Iberian Peninsula while group B is found everywhere else. Domestic breeds belong to the latter. As evidenced from data on ancient bones (up to 12,000 years BP) the mitochondrial type B1, predominant in domestic animals, originated from Spain. B1 animals were introduced in France by man between late Roman times and Middle Ages. PMID- 9404457 TI - [Genetics of aging]. AB - The classical arguments favouring a genetic basis of aging are reviewed emphasizing the questions that remain unanswered. Genes cannot be the sole genetic determinants of aging. Mendel's paradigm cannot anymore explain all the results recently obtained. Different aspects of the organization of the genome must also play a role in aging. The functions of the largest part of the human genome remain unknown. Moreover the different genetic theories of aging are based on natural selection. However, other paradigms are being proposed that can complement or replace Darwin's paradigm. They are based on the spontaneous organization of complex systems. They must be considered in the reappraisal of the aging phenomenon. PMID- 9404458 TI - [Genetics of longevity]. AB - It is possible to use different studies to demonstrate the genetic component of a phenotype. From the beginning of the century, many authors have studied the possible genetic transmission of longevity. The study of genealogies of ascendants first, and then of descendants of elderly individuals shows that the age at death incorrelated in people belonging to the same family. Finally, the studies carried out on monozygot and dizygot twins have made it possible to estimate that this genetic component accounts for approximately 10% of the individual's lifespan. Research on congenic mice which differ only by the chromosomic region of the Major Histocompatibility Complex (MHC) implies that this particular region might have an effect on longevity. However, the study of several alleles of the MHC indicates a complex sex-dependent influence. Several other chromosomic regions are also implied. As far as human beings are concerned, several research teams have worked on the HLA region. Once again, the situation is still far from clear. PMID- 9404459 TI - [Genetic analysis of two cellular degenerations in filamentous fungus Podospora anserina]. AB - The filamentous fungus Podopsora anserina presents an unavoidable arrest of vegetative growth (Senescence) determined by a cytoplasmic and infectious factor. Senescence is correlated with a disorganization of the mitochondrial DNA. This disorganization is caused by an event which is not the appearance of the first defective DNA molecules. These ones are generated constitutively and their accumulation during Senescence requires the presence of an additional factor. Life span of the strains is under nuclear and cytoplasmic genetic control. At least 600 nuclear genes influence longevity. Our analysis focuses on the role of the genes involved in cytosolic translation, since mutations in these genes seem to display the most drastic effects on longevity but also on the structure of the defective mitochondrial DNA molecules that accumulate during Senescence. We have detected in some Podospora anserina mutant strains (permissive strains) the presence of a novel cytoplasmic and infectious determinant that entails an easily discernible phenotype associated with a severe growth alteration (Crippled Growth). This growth alteration is not associated with mitochondrial DNA modifications. Only the strains that have an increased translational accuracy present Crippled Growth. However, the Crippled Growth Determinant is found in all the strains during the stationary phase; it is eliminated from the non permissive strains during the exit of the stationary phase. The mutants, that have an increased translational accuracy, probably lack a factor which is needed to eliminate the determinant when cells enter the growth phase. PMID- 9404460 TI - [Do mitochondria play a role in aging?]. AB - Ageing is an unavoidable and complex phenomenon which may be a price to pay to evolution. Thus genetics appear to play a predominant role besides environmental factors. Energetic metabolism slowly declines with ageing supporting a possible active role of mitochondria, the power supply of the cells, to this process. Mitochondrial DNA alterations appear during the mid-life and in degenerative diseases such as in Parkinson's and Alzheimer's; they include large scale deletions and point mutations. Since the respiratory chain plays a major role in the generation of superoxide anions which are converted into hydroxyl radicals that may impair lipids, proteins and DNA function in mitochondria, this vicious cycle may result from both an altered control of mitochondrial biogenesis dependent from the nucleus, and/or from a lack of repair and accumulation of somatic mitochondrial DNA mutations. PMID- 9404461 TI - [Relation between genetic and epigenetic mechanisms in aging]. AB - The recent identification of genes involved in the regulation of the longevity of some invertebrates (drosophila, nematodes) as well as the presence of human genes exhibiting homologies to those identified in invertebrates, revived the deterministic theories of aging. It appears however that those mechanisms which were shown experimentally to be involved in aging in vertebrates and in particular in humans belong to the epigenetic mechanisms such as the Maillard reaction and free radical attack in particular. Some diseases which imitate an accelerated aging as Progeria and the Werner syndrome are due to mutations, some of them mutagenic themselves. Reflections based on such arguments concerning reactions with catalysers coded in the genome but with activities escaping strict control, suggest an indirect determinism of phenomena involved directly in cell- and tissue aging. These considerations are illustrated by the example of the elastin-laminin receptor studied in our laboratory. Its sustained activation by circulating elastin peptides appears to be involved in cell and tissue aging. PMID- 9404462 TI - [Oxidative stress and chronic renal insufficiency: what can be a prophylactic approach?]. AB - Cardiovascular diseases represent the first cause of mortality in chronic renal failure patients treated by hemodialysis. Alterations in lipid metabolism and oxidative stress are recognized as vascular risk factors. Their corrections could be of interest for atherosclerosis prevention. In order to evaluate interest of an therapeutic intervention, we have analyzed oxidative metabolism in hemodialysis patients by determining the production of oxygen reactive species (ROS), the level of defense mechanisms, and the balance between nitric oxide (NO) and ROS, responsible for anti- or proxidant effects of NO. During dialysis sessions performed with cellulosic membrane (Cuprophan) an increase in hydroperoxide production by platelets was noted (12 HETE) (5.62 +/- 0.94 pg); similarly, superoxide anion (O2(0)-) production by monocytes (fluorescence index: 115 +/- 24) and by polynuclear cells (fluorescence index: 115 +/- 24) was enhanced. On the other hand, anti-oxidant defenses were significantly reduced with a decrease in RBC SOC activity (0.92 +/- 0.06 U/mg Hg) and in RBC vitamin E (0.7 +/- 0.07 mg/l) concentration. We have demonstrated a profound alteration in the L-arginine/NO pathway consequently to an accumulation of NO synthases inhibitors or activators. The necessity to reduce the production of ROS during dialysis sessions justifies the use of more biocompatible membranes, such as modified cellulosic or synthetic membranes, decreasing leucocyte activation. In addition, NO synthetase inhibitors can be preferentially eliminated by convection. Finally, a supplementation with an exogenous anti-oxidant, such as oral vitamin E (500 mg/day for 6 months) normalizes RBC vitamin E levels and concomitantly allows a decrease in MDA concentrations In conclusion, oxidative metabolism alterations observed in hemodialysis are multifactorial: preventive measures include the use of a more biocompatible material, the reequilibrium of the NO/ROS balance, and supplementation with exogenous anti-oxidants. PMID- 9404464 TI - [In vivo and in vitro study of the effect of somatostatin and somatostatin immunoreactive substance on the histology of the mantle edge in adult and juvenile snails (Helix aspersa)]. AB - We have demonstrated structural differences between the mantle edge gland of young, adult and adult shell repairing snails. In vivo and in vitro, somatostatin 14 and somatostatin-immunoreactive substance (SSI) induce a juvenile status of the gland in adults and an activation of juvenile ones. These results suggest that they could regulate the growth and the repair of the shell and that the mantle edge gland might be considered as one of their tissue targets. PMID- 9404463 TI - [Modulation of respiratory activity of renal macrophages in sea bass (Dicentrarchus labrax) by chronic exposure to sublethal concentration of ammonia]. AB - Sea bass (Dicentrarchus labrax) were exposed for 71 days to three different ammonia concentrations corresponding to 0, 15 and 25% of the lethal threshold concentration causing the death of half a fish population in 96 hours. The study of the respiratory burst from sea bass renal macrophages showed that the luminol dependent chemiluminescence (CL) emitted following stimulation with mezerein is higher when fishes were previously exposed to ammonia. Therefore, it seemed that chronically exposure of fishes to sublethal concentrations of ammonia primed their renal macrophages to secrete higher amounts of oxygen activated species during respiratory burst, even several days after the transfer of fishes into a standard environment. The stimulated-macrophage CL was partially inhibited by sodium azide, superoxide dismutase and a nitric oxide-synthesis inhibitor, the N5 (-1-iminoethyl)-L-ornithine monochloride, showing that hydrogen peroxide, superoxide anions and nitric oxide were released by renal macrophages from sea bass during the respiratory burst. These reactive species could react together to generate peroxynitrite, a strong bactericidal agent. PMID- 9404465 TI - [Dynamic phototherapy: study of anti-tumour potentialities of bis (tri n hexylsiloxy) silicon phthalocyanine on malignant achromic M6 melanocytes. EPR study of phototoxic mechanism]. AB - Photodynamic therapy of cancerous cells is a technique relying upon the irradiation of tumorous cells after selective incorporation of a photosensitizer. The bis (tri n-hexylsiloxy) silicon phthalocyanine is a second generation photosensitizer. Anti-cancerous potentialities of this molecule have been evaluated against the melanotic M6 cell line. Results have evidenced a high phototoxicity at low concentration and no dark toxicity under the same conditions. EPR studies on the photochemical pathways involved in phototoxicity processes have been realised in solvent and model membranes (liposomes). Results provide evidences for the production of singlet oxygen (1O2) as well as superoxide (02(o)-) and hydroxyl radicals (0OH). PMID- 9404466 TI - Is conventional radiography suitable for evaluation of a disease-modifying drug in patients with knee osteoarthritis? PMID- 9404467 TI - Elevated aggrecan mRNA in early murine osteoarthritis. AB - Male STR/ort mice develop osteoarthritis in the tibial articular cartilage. Low grade histological lesions first appear between 10-20 weeks of age. A previous study showed that the level of aggrecan, the major cartilage proteoglycan, is approximately twofold greater in the tibial cartilage of 16-19-week-old STR/ort mice compared with that in normal cartilage in control CBA mice. In the present investigation aggrecan gene transcription was investigated in 20-week-old STR/ort and CBA tibial cartilage using a quantitative reverse transcription-polymerase chain reaction (RT-PCR). The amount of aggrecan cDNA obtained from the STR/ort medial and lateral plateau was 2.8- and 4.6-fold greater per milligram of wet cartilage than that from the CBA tibial plateau. The difference was not due to differences in cellularity of tibial cartilage in the two strains and indicates that aggrecan gene transcription is elevated in early osteoarthritis. PMID- 9404468 TI - Hyaluronic acid suppresses fibronectin fragment mediated cartilage chondrolysis: II. In vivo. AB - Intra-articular sodium hyaluronic acid (HA) has been used as a treatment intervention in the management of osteoarthritis. It has been observed that HA can coat the articular surface, and thus, has been suggested to provide a possible prophylactic barrier for the articular cartilage. In an accompanying manuscript (Homandberg et al.), we report that a commercially available high molecular-weight HA (approximately 800-kDa, ARTZ, Seikagaku Corp.) can partially block fibronectin fragment (Fn-f)-mediated cartilage injury in vitro. Herein we report a study of the effects of intra-articular HA on an in vivo animal model of Fn-f-mediated cartilage injury. Rabbit knees were injected with Fn-f, and after 1 week, the cartilage proteoglycan (PG) content had decreased to 59 +/- 8% of control. In sharp contrast, PG content in knees receiving pre-treatment with HA followed by Fn-f injection had only decreased to 85 +/- 27% of control (P < 0.01). Similarly, the PG content in knees receiving an injection of Fn-f, followed by an injection of HA were significantly higher (74 +/- 18% of control) than Fn-f injured knees with no treatment (P < 0.02). Intra-articular HA alone had no effect on cartilage PG content. The results in this study suggest that HA is effective in partially preventing Fn-f mediated cartilage injury, most likely by coating the articular surface. Further, HA treatment after Fn-f injury may facilitate restoration of matrix components. PMID- 9404469 TI - Defining radiographic osteoarthritis for the whole knee. AB - OBJECTIVE: To determine in the knee which individual radiographic feature or combination of features in the patellofemoral and tibiofemoral joints correlate best with a nonradiographic definition of clinical osteoarthritis in order to recommend a definition of radiographic osteoarthritis for use in studies. METHODS: Using data from the Framingham Osteoarthritis Study, we tested the correlation of clinical OA, defined as frequent knee pain plus crepitus, with a variety of definitions of radiographic OA including those based on individual radiographic features, e.g. > or = grade 2 osteophyte 0-3 scale, and new definitions that included alternative combinations of features, [e.g. either > or = grade 2 osteophyte or joint space narrowing > or = grade 2 (0-3 scale) with a bone feature (such as cyst, sclerosis, or grade 1 osteophyte)]. We performed analyses looking at participants who had obtained both weight-bearing anteroposterior (AP) and lateral radiographs of both knees. RESULTS: In 519 participants, we found that the definitions of radiographic osteoarthritis best correlated with clinical OA were definite osteophyte > or = grade 2' (efficiency 62.4-67.1%) and an 'alternate definition' of either osteophytes > or = grade 2 or joint space narrowing > or = grade 2 with a bony feature of OA (efficiency 62.8 68.1%). A recursive partitioning analysis selected the 'alternate definition' as best. Also, we found that adding lateral views to the AP view improved the diagnostic test performance of the best performing radiographic definitions. CONCLUSION: We suggest that a knee should be characterized as having radiographic OA if there is either an osteophyte of grade 2 or greater severity (0-3 scale) present or the presence of moderate to severe joint space narrowing (> or = 2 on a 0.3 scale) with co-occurrence of a bony feature in the compartment affected. PMID- 9404470 TI - The effects of hyaluronan during the development of osteoarthritis. AB - Ninety-nine mature New Zealand White (NZW) rabbits underwent unilateral anterior cruciate ligament transection (ACLT) and were divided into three groups. The contralateral non-operated knees served as controls. The first group (SA) received intra-articular injections of 0.3 ml hyaluronan (HA: MW; 8 x 10(5)) beginning 4 weeks after ACLT, once a week for 5 weeks. The second group (SV) was injected with vehicle (carrier of HA) in the same fashion as the SA group. The third group (SN) served as a nontreatment group post ACLT. All animals were killed 9 weeks post-surgery and were assessed by gross morphology, histomorphometry and biochemical analysis. Gross morphologic changes on the femoral cartilage in the SA group were less severe than those in the SV and SN groups. Cartilage thickness, cartilage area, and thickness of synovial lining cell layer histomorphometric parameters were measured, showing a positive effect of HA on the preservation of articular cartilage and synovial tissue. Similarly, the cartilage and synovial tissues from knees injected with HA did not demonstrate significant alterations from contralateral controls as measured by biochemical analysis [i.e., water content, pyridinoline concentration, glycosaminoglycan (GAG) content for the cartilage, and DNA concentration for the synovial tissue]. PMID- 9404471 TI - Isolated chondrons: a viable alternative for studies of chondrocyte metabolism in vitro. AB - OBJECTIVE: To develop and test a simple enzymatic procedure for isolating chondrons, which consist of the chondrocytes and their surrounding pericellular microenvironment. DESIGN: Chondrons were obtained by digesting adult human articular cartilage with a mixture of dispase and collagenase. Chondrons and chondrocytes were cultured in alginate beads, immunofluorescence labeled and examined by confocal microscopy. RESULTS: Comparison of freshly isolated chondrons with cryostat sections of cartilage revealed that type VI collagen, type II collagen and aggrecan were retained, but fibronectin and a unique chondroitin sulfate epitope recognized by the antibody, 7D4, were lost. Comparison of enzymatic and mechanical homogenization methods revealed subtle changes in chondron morphology and retention of fibronectin in mechanically isolated chondrons. Average yield of enzyme-isolated chondrons was slightly lower than that of chondrocytes isolated by pronase and collagenase digestion, but was much greater than that reported for mechanically isolated chondrons. Enzyme isolated chondron viability was greater than 80% 1 day after isolation, and continued to be above 80% through 7 weeks of alginate bead culture. Viability of isolated chondrocytes was initially greater than 80% but fell to 60-80% with time in culture. Chondrons and isolated chondrocytes had a similar division rate except osteoarthritic chondrons were significantly slower after 2 weeks in culture. Cell division was more rapid for nonosteoarthritic chondrons than for osteoarthritic ones. CONCLUSIONS: Enzymatic isolation of chondrons is relatively simple, gives better yield and viability than mechanical isolation, but comparable yield and viability of traditional chondrocyte isolation. Enzymatic chondron isolation allows the effect of the in vivo-formed pericellular matrix on chondrocyte metabolism to be studied in vitro. PMID- 9404472 TI - Transplantation of adenovirally transduced allogeneic chondrocytes into articular cartilage defects in vivo. AB - Gene transfer to chondrocytes followed by intra-articular transplantation may allow for functional modulation of chondrocyte biology and enhanced repair of damaged articular cartilage. We chose to examine the loss of chondrocytes transduced with a recombinant adenovirus containing the gene for Escherichia coli beta-galactosidase (Ad.RSVntlacZ), followed by transplantation into deep and shallow articular cartilage defects using New Zealand White rabbits as an animal model. A type I collagen matrix was used as a carrier for the growth of the transduced chondrocytes and to retain the cells within the surgically created articular defects. Histochemical analysis of matrices recovered from the animals 1, 3 and 10 days after implantation showed the continued loss of lacZ positive chondrocytes. The number of cells recovered from the matrices was also compared with the initial innoculum of transduced cells present within the matrices at the time of implantation. The greatest loss of transduced cells was observed in the first 24 h after implantation. The numbers of transduced cells present within the matrices were relatively constant between 1 and 3 days postimplantation, but had progressively declined by 10 days postimplantation. These results suggest that transduction of chondrocytes followed by intra-articular transplantation in this rabbit model may enable us to examine the biological effects of focal transgenic overexpression of proteins involved in cartilage homeostasis and repair. PMID- 9404473 TI - A double-blind, randomized trial to compare meloxicam 15 mg with diclofenac 100 mg in the treatment of osteoarthritis of the knee. AB - Meloxicam is a new nonsteroidal anti-inflammatory drug (NSAID), which, in animal tests, displays a high potency for anti-inflammatory and analgesic action. The aim of this study was to investigate the efficacy and tolerability of 15 mg meloxicam in comparison with 100 mg slow-release diclofenac in patients with osteoarthritis of the knee. Two hundred and fifty-eight patients were included in the intent-to-treat analysis; these were randomized into two groups to receive either 15 mg meloxicam (N = 128) or 100 mg diclofenac (N = 130) for a period of 6 weeks. The results with respect to efficacy showed a trend in favor of meloxicam regarding pain on movement, global efficacy and paracetamol consumption, although these differences did not reach statistical significance. The most frequently occurring adverse events in both groups were of a gastrointestinal (GI) nature. However, there was a higher incidence (26 vs 16%) of GI adverse events in the diclofenac group compared with the meloxicam group. Both drugs were well tolerated when assessed by the patients on a visual analog scale (VAS). Thus, 15 mg meloxicam is an effective and well-tolerated therapy for osteoarthritis and compares favorably with diclofenac 100 mg, a well-established treatment for this indication. PMID- 9404474 TI - Effect of diacetylrhein on the development of experimental osteoarthritis. A biochemical investigation. PMID- 9404475 TI - [Myelodysplastic syndromes]. PMID- 9404476 TI - [Etiological factors of myelodysplastic syndromes]. AB - Specific epidemiologic data on myelodysplastic syndromes are rare. Analysis of data is in fact affected by problems of terminology and classification. The link between the exposure to ionizing radiation or alkylating agents and MDS is well established. Etiologic factors of acute leukemia, or new factors such as non ionizing radiation, solvent, ethylene oxide, glycol eters, tobacco smoke, exhaust gases, agricultural work have been hypothesized but should be confirmed by other studies on MDS. PMID- 9404477 TI - [Cytologic aspects and nosographic update of myelodysplastic syndromes]. AB - A discussion is provided of the cytologic abnormalities in myelodysplastic syndromes (MDS), which remain central to the diagnosis and classification of these conditions. Accurate identification of the blast cells is essential since the proportion of these cells is of considerable prognostic significance. A description is given of the FAB classification scheme, which is both simple and reproducible. Prognostic scoring systems based on bone marrow blast proportion, cytopenias, and karyotypic abnormalities are also discussed. PMID- 9404478 TI - [Cytogenetics of myelodysplastic syndromes]. AB - The authors review the main cytogenetic abnormalities encountered in MDS particularly del 5q, -7, +8, and complex abnormalities. Their presence may be useful to the diagnosis of MDS in difficult cases. Their prognostic value is important, and cytogenetics have recently been included in prognostic scores in MDS. Knowledge of recurrent chromosomal rearrangements is finally a first step in the discovery of genes implicated in the pathogenesis of MDS. PMID- 9404479 TI - [Molecular abnormalities and clonality in myelodysplastic syndromes]. AB - Few genes have a proven role in the pathogenesis of myelodysplastic syndromes (MDS). The most common abnormalities involve the RAS genes, most notably the N RAS gene, and are present in 10% of cases at diagnosis and in 30% to 40% during the course of the disease. Mutations of the p53 are found in 5% to 10% of cases. Mutations of the cFMS genes are less common, abnormalities of the NF1 genes seem to occur only in children, and abnormalities of the RB genes are exceedingly rare. A few instances of t(5;12) or t(3;21) translocation have been demonstrated, and their study has provided evidence that the TEL, EVI1, MDS1, and AML1 genes are involved in some cases of MDS. The presence in MDS of recurrent chromosome 7, 5q, and 20q deletions suggests that these chromosomal segments may bear tumor suppressor genes involved in MDS. The gene(s) involved remain(s) to be identified. Clonality studies have shown that stem cell involvement usually occurs at the myeloid level and that normal multipotent stem cells persist in many patients with MDS. This opens up the promising possibility that transplantation of autologous multipotent stem cells may be an effective therapeutic approach. PMID- 9404480 TI - RAS and the myelodysplastic syndromes. AB - RAS genes have been implicated in several different malignancies. The mechanism of activation in most cases has been due to point mutations at critical domains responsible for guanine nucleotide binding. These changes alter the conformation of the protein resulting in insensitivity of the protein to the GTPase activating protein which normally hydrolyses the active p21RAS GTP-bound form to the inactive GDP-bound form. RAS genes have potent effects on the differentiation and proliferation program of cells. The mechanism induced depends on the context in which RAS is found as well as its mutational status and indeed which RAS gene family member is involved. RAS mutations have been described early in the disease process in haematologically normal individuals at risk of mutations induced by either occupational hazard exposure, such as benzene, or of secondary disease after chemotherapy for a previous malignancy. It also been associated with disease progression from myelodysplasia (MDS) to acute myelogenous leukaemia (AML), but it has also been described to be lost upon disease progression, thus showing that RAS mutations are unlikely to be initiating events or at least not required for maintenance of disease. As RAS appears to be involved in primary and secondary myeloid leukaemias, it is a good candidate for gene targeted therapeutic intervention. Studies to target RAS either directly or indirectly by interfering in the RAS pathway are underway. Clinical trials with a peptide RAS vaccine are also ongoing in solid tumours. This report seeks to review the evidence for RAS involvement as oncogenes, focusing on MDS, the reasons as to why the hot spots of codons 12/13 and 61 are particularly potent in activating the transformation potential of RAS and the different approaches being undertaken to translate laboratory findings into therapeutic reality. PMID- 9404481 TI - [Abnormalities of the expression of cytokines and their receptors in myelodysplastic syndromes]. AB - Abnormalities of the expression of cytokines and their receptors seen in myelodysplastic syndromes (MDS) are reviewed. Autocrine and paracrine secretion of growth factors, most notably GM-CSF and IL-6, has been identified as a factor in clone expansion in young pediatric patients with chronic myelomonocytic leukemia. Mutations affecting cFMS, the receptor for M-CSF, have also been demonstrated. The other reported abnormalities have been found in too small a number of cases to explain the cell growth and differentiation disorders seen during the course of MDS. It remains possible, however, that a more detailed study of signal transduction pathways might detect functional abnormalities explaining the contrast between the small number of reported cases with abnormalities at the cytokine and cytokine receptor sites and the cell growth and differentiation disorders that are prominent in MDS. PMID- 9404482 TI - Apoptosis as a cell biological abnormality in myelodysplasia. AB - Evidence is accumulating to indicate that alterations in the control of apoptosis can contribute to a variety of disorders. Among the disorders associated with abnormal regulation of apoptosis, MDS stands out unequaled in that apoptosis is related, in apparently opposite directions, to ineffective hematopoiesis and leukemic transformation of MDS; notably, excessive apoptosis in the former and deranged apoptosis or escape from apoptotic control in the latter. In this review, we want to focus on the role of apoptosis in various stages of MDS, and to discuss its possible relevance to further therapeutic interventions. PMID- 9404483 TI - [Myelodysplastic syndromes: unusual and mild forms]. AB - Although the FAB classification of the myelodysplastic syndromes (MDS) allows to classify most patients, a few clinical patterns do not fit the FAB categories, including borderline forms with manifestations usually seen in other diseases and forms with atypical or unusual clinical or laboratory features that do not immediately suggest a MDS. Borderline forms are characterized by the presence of any of the following: high or very high platelet counts, myelofibrosis, bone marrow hypoplasia, eosinophilia, or systemic diseases such as relapsing polychondritis. Unusual or atypical forms include manifestations such as hemolysis, high reticulocyte counts, erythroblastopenia, or an abnormality of a single cell line such as isolated thrombocytopenia, isolated neutropenia, or isolated macrocytosis. The definitive diagnosis of these forms of MDS can require a number of investigations such as cytogenetic studies, bone marrow biopsy, and/or radionuclide evaluation, and may not be possible until the patient has been followed for some time. PMID- 9404484 TI - Atypical chronic myeloid leukemias. AB - An ideal classification of the chronic myeloid leukemias would be based on a thorough understanding of the aetiology, pathogenesis, clinical and laboratory features, and natural history of the various conditions which comprise CML. Only in the case of CGL is the pathogenesis well understood and the diagnosis of the remaining disorders is still largely based on clinical and morphological criteria. It is inevitable for the reasons previously discussed that there will be cases which either defy classification or fall within the diagnostic criteria of more than one disorder. As long as careful clinical and morphological observation continues in parallel with advances in cellular and molecular biology it seems inevitable that the current debate about the inter-relationship between CMML, aCML and Philadelphia negative CGL will be resolved. PMID- 9404485 TI - [Myelodysplasia in children and mitochondrial cytopathies]. AB - Sideroblastic anemia associated with vacuolization of haemopoietic precursors can be observed in some constitutional diseases associated with mitochondrial DNA deletion. In this condition, it is the haematological expression of a multi tissue disorder. Haemopoiesis is polyclonal, without abnormality of nuclear DNA differing from the acquired idiopathic sideroblastic anemias which arise from a clonal transformed stem cell. This model of childhood polyclonal myelodysplasia can be observed in others myelodysplasias associated with constitutional polymalformative syndromes. PMID- 9404486 TI - Juvenile chronic myelomonocytic leukemias: molecular and novel therapeutic basis. AB - Myelomonocytic leukemias of children represent a distinct disorder from chronic myelogenous leukemia and adult myelomonocytic leukemias. The prognosis is poor and no therapy has proved to be yet effective. This review summarizes the molecular and cellular characteristics of this disease which may allow to design more targetted therapeutic agents. PMID- 9404487 TI - [Chronic myelomonocytic leukemia in adults]. AB - Chronic myelomonocytic leukemia (CMML) is characterized by the association of myelodysplastic features and proliferation of both granulocytic and monocytic series. This disease frequently indolent, may however demonstrate progressive course and/or transform to acute leukemia. Nosologic and diagnostic problems raised by this malignant hematological pathology, its clinical and biological presentations, and current treatments are reported in this review. PMID- 9404488 TI - Contextual effects on colour appearance: lightness and colour induction, transparency, and illumination. PMID- 9404489 TI - Lightness and junctions. AB - The lightness of a test patch completely surrounded by an inducing field can be predicted by variants of Wallach's ratio rule. When a patch is surrounded by two or more regions with different luminances, a plausible extension of the ratio rule would predict that the effect of the surrounding regions should correlate with the length of the border they share with the test patch. However, as shown by the Wertheimer-Benary and White effects, lightness of such patches can depart appreciably from these predictions. It is argued that a fruitful approach toward the explanation of such effects is based on the analysis of junctions (such as T junctions and X-junctions) between regions. Several new displays and variations of old displays involving such junctions are used to illustrate this approach. An alternative analysis of a lightness effect introduced by Adelson is provided, and the role of depth effects in achromatic perception is discussed. A number of limitations of the approach and possible ways to overcome them are noted. PMID- 9404490 TI - Induced effects of backgrounds and foregrounds in three-dimensional configurations: the role of T-junctions. AB - In three-dimensional configurations, and two-dimensional pictures of such configurations, simultaneous contrast induction from proximate backgrounds affects perceived brightness, color, and internal contrast to a greater extent than induction from coplanar or occluding surrounds or from more distant backgrounds. In the projected image the presence of occluding flanks or retinally adjacent distant backgrounds is indicated by T-junctions. However, the presence of T-junctions inhibits induced contrast irrespective of the three-dimensional percept. The configurations in this paper refute the notions that perceived coplanarity or perceptual belonging necessarily enhance induced contrast. PMID- 9404491 TI - New configurational effects on perceived contrast and brightness: second-order White's effects. AB - Novel phenomena of perceived contrast and brightness in spatial configurations are presented, which are of the type used by White but consist exclusively of contrast variations or of combined contrast and luminance variations. As with White's effect, perceived contrast and brightness in these displays do not correspond to predictions based on low-level mechanisms of contrast and brightness induction, and it is suggested that spatial organisation influences the appearance of first-order and second-order stimuli in a similar fashion. PMID- 9404492 TI - A theory of illusory lightness and transparency in monocular and binocular images: the role of contour junctions. AB - A theory of illusory transparency and lightness is described for monocular and binocular images containing X-, T- and I-contour junctions. This theory asserts that the geometric and luminance relationships of contour junctions induce illusory transparency and lightness percepts by causing a phenomenal scission of a homogenous luminance into multiple contributions. Specifically, it is argued that a discontinuous change in contrast along aligned contours that preserve contrast polarity induces a scission of the lower contrast region into a near transparent surface or an illumination change, and a more distant surface that continues behind behind this near layer. This scission is assumed to cause changes in perceived lightness and/or surface opacity. Discontinuous changes in contrast along contours also are assumed to induce end-cut illusory contours that run roughly perpendicular to the inducing orientation of the contour, both monocularly and binocularly. Binocular illusory contours are shown to be caused by the presence of unmatchable contour terminators. It is argued that the presented theory can provide a unified account of a variety of monocular and binocular illusions that induce uniform transformations in perceived lightness, including neon-color spreading, the Munker-White illusion, Benary's illusion, and illusory monocular and binocular transparency. PMID- 9404493 TI - A vector model of colour contrast in a cone-excitation colour space. AB - A vector model of colour contrast is examined in a colour space that is a logarithmic transformation of the MacLeod-Boynton cone-excitation diagram. Observers set matches in a haploscopic display, in which one eye viewed a standard display (a neutral target square in a coloured surround) and the other viewed a matching display (a variable square in its own surround). Contrast colours are simply represented in this colour space: the vector connecting the right-eye surround and matched chromaticities is parallel to and to the same length and direction as the vector that connects the left-eye (standard) surround and square chromaticities. This describes observers' matches to the hues induced in a neutral square for a range of inducing surround colours, a range of right eye (match) surround colours and four different luminance contrasts. PMID- 9404494 TI - Color transparency. AB - Observation suggests that the chromatic changes which elicit an impression of transparency include translations and convergences in color space. Neither rotations nor shears in color space lead to perceived transparency. Results of matching experiments show that equiluminous translations, which cannot be generated by episcotister or filter models, give rise to the perception of transparency. This implies that systematic luminance change is not needed for transparency to be perceived. These results were used for the development of a method for detecting a transparent overlay within a color image and for separating the overlay from the underlying surfaces. The method tests for the coherence of chromatic change along contours through X-junctions to help detect the contour of a transparent region. The algorithm tests locally for translation and convergence to detect a transparent region. It estimates globally the chromatic parameters of the transparent overlay in order to separate the overlay from the underlying surfaces. PMID- 9404495 TI - Brightness with and without perceived transparency: when does it make a difference? AB - Subjects matched the brightness of test patches whose inner (adjacent) surrounds appeared either as transparent overlays on a wider background that included the test patch or as regions differing in reflectance from the test patch and the outer surround. In the above configurations the luminance and spatial extent of the inner surround was identical, thus controlling for the effects of surround luminance. Configuration condition had a significant effect on test-patch brightness. In general, test-patch brightness was significantly elevated under conditions favouring the interpretation of the stimulus as including a transparent overlay. The largest effect occurred for the configuration in which the perception of transparency was supported by stereo depth cues. The brightness effect was mediated by the virtual transmittance of the transparent overlay, increasing in magnitude with decreasing transmittance. Further, the effect of transparency on brightness was greatest for test-patch luminances near to those of their immediate surrounds. PMID- 9404496 TI - An account of brightness in complex scenes based on inferred illumination. AB - Achromatic brightness matches between two small patches were measured in a display containing ten larger regions of different luminances. The spatial organization of the ten regions was varied while keeping constant the immediate surround (and thus local contrast) of each patch as well as the average luminance of the entire stimulus. Various spatial arrangements were designed to alter the illumination inferred by the observer without changing the ensemble of luminances actually in view. Some spatial arrangements of the ten regions were consistent with five (simulated) surfaces under two distinct levels of illumination, with one luminance edge within the display (an 'apparent illumination edge') dividing the stimuli into an area of lower illumination and an area of higher illumination. In other spatial arrangements the ten regions were configured so that no luminance edge in the display could be interpreted as an ecologically valid illumination edge that provides a parsimonious interpretation of the ten regions; these conditions were designed to induce observers to infer ten surfaces under a single illuminant. When the ten regions were arranged with an apparent illumination edge, the patch within the area of lower perceived illumination was perceived as dimmer than when the same patch and immediate surround were presented with no apparent illumination edge. The results are interpreted by positing that the apparent illumination edge causes an observer to group together regions under the same perceived illuminant, with a consequent effect on brightness: lowering or raising the level of a perceived illuminant causes a patch of fixed contrast to be perceived as less bright or more bright, respectively, just as occurs when lowering or raising the level of real illumination. It is suggested that changes in brightness in a complex scene that result from a change in real illumination may be caused by a difference in inferred illumination at the perceptual level, not by simply a change in the amount of light absorbed by photoreceptors. PMID- 9404497 TI - Shading and stereo in early perception of shape and reflectance. AB - Recent experiments involving shaded 2-D stimuli have shown that early-vision mechanisms are capable of interpreting 3-D shape from shading. In particular, target discrimination tasks suggest that a target 'pops out' when background distractors, but not the target, can be interpreted as convex and lit from above or top-left. Since the problem of extracting 3-D shape from shading is intrinsically ill-defined, early vision may need to make these twin assumptions of convexity and top-left lighting in order to constrain the problem. Would these assumptions be recognized as unnecessary and consequently discarded when 3-D shape could be unambiguously defined by some other cue, like stereo disparity? A 2AFC stimulus onset asynchrony paradigm with masking was used in target discrimination experiments. The performance of five naive subjects on tasks where only shading cues were present was compared with that on tasks involving shading as well as stereo cues that define shape unambiguously. The results show that although stereo disparity information is incorporated by early-vision 3-D mechanisms, it is not used to overturn the default assumptions of lighting and shape. Stereo information is interpreted within the framework of top-left lighting, and a consistent preference for convexity is seen over concavity. PMID- 9404498 TI - Ambiguities in colour constancy and shape from shading. AB - A new visual phenomenon--called the AMBEGUJAS phenomenon--is presented, together with some descriptive data from two initial exploratory experiments. The phenomenon is basically one of shape from shading, but ambiguous as to both shape and colour. There are two spontaneously alternating and mutually exclusive perceived 3-D shapes, and--as the most surprising observation--the colour impressions of these two shapes are markedly different. The stimulus situation is very simple with two differently coloured illuminations (with sharp edges) adjacently cast onto a flat, grey striped surface. In one 3-D shape almost the whole chromatic content disappears, and the surface goes towards its veridically grey colour. In the other the perceived object assumes the two illumination colours as clear surface colours. The decolorised percept is interpreted as a striking example of colour constancy: a perceptual solution with the classical 'discounting of the illuminant'. Experiments show that the phenomenon is robust and appears in varying display layouts and different combinations of chromatic illuminations. PMID- 9404499 TI - Spore coat differentiation in Bacillus subtilis. PMID- 9404500 TI - Inhibition of bacterial cell surface extension by various means causes blocking of macromolecular synthesis. AB - It has been suggested that, in rod-shaped bacteria, two sites for peptidoglycan assembly exist: one which is responsible for septum formation and the other, for lateral wall extension. The balance between the activities of these two sites enables bacteria to conserve their own morphology during cell growth. The effect of specifically inhibiting septum formation by different means (antibiotics and/or mutations), upon cell surface extension and macromolecular synthesis in rod-shaped and coccoid bacteria of various species, was studied. Inhibition of either cell wall expansion or macromolecular synthesis did not occur when septum formation was impaired in both rod-shaped bacteria and cocci possessing the two sites for peptidoglycan assembly, whereas a rapid and complete block of such synthesis was caused by inhibiting both sites in rod-shaped bacteria, or septum formation in cocci which possess only this site. These data indicate that bacteria possess a control mechanism that prevents macromolecular synthesis when envelope extension is inhibited. PMID- 9404501 TI - Molecular characterization of the Salmonella typhi StpA protein that is related to both Yersinia YopE cytotoxin and YopH tyrosine phosphatase. AB - Analysis of the nucleotide sequence of a 4-kb DNA fragment located between the sip and iag loci on Salmonella typhi chromosome revealed three open reading frames, termed sipF, ctpA and stpA. The 82-amino-acid (aa) sipF product showed extensive similarity to the lacP protein from S. typhimurium. The StpA protein (535 aa) exhibited significant similarity to both Yersinia enterocolitica YopE cytotoxin and YopH tyrosine phosphatase. The CtpA polypeptide (130 aa) might be the molecular chaperone of the StpA protein. PMID- 9404502 TI - Glucose starvation response in Enterococcus faecalis JH2-2: survival and protein analysis. AB - We investigated the survival of Enterococcus faecalis following starvation provoked by energy source glucose exhaustion. Inhibition of protein synthesis by chloramphenicol before 3 h of starvation resulted in a dramatic decrease in viable bacteria. Antibiotic treatment of cells after 3 or 6 h of starvation had a progressively lesser influence on bacterial survival. During the first 24 h of deprivation, a total of 42 proteins were identified as glucose-starvation inducible; 4 temporal classes of proteins (A, B, C and D) were defined in relation to their enhanced synthesis after glucose exhaustion. Our results show that proteins from the two early classes (A and B) seem to be the most important for long-term survival in E. faecalis. One protein of each of these classes was analysed at the molecular level. The N-terminal sequence of one of them, belonging to class A, showed strong homology with the N-terminal sequence of carbamate kinase from Streptococcus faecium. This enzyme could be implicated in the development of alternative metabolic pathways of energy production and could be compared to the Cst proteins of Escherichia coli. PMID- 9404503 TI - Oxygen-dependent upregulation of transcription of alginate genes algA, algC and algD in Pseudomonas aeruginosa. AB - The mRNA levels of algA, algC and algD genes increased, coordinately, in cells of the highly mucoid Pseudomonas aeruginosa 8821M grown under increasing dissolved oxygen tensions (DOT) of up to 70% of air saturation. These genes encode the bifunctional protein with phosphomannose isomerase (PMI) and GDP-mannose pyrophosphorylase (GMP) activities (algA), the phosphomannomutase (PMM) (algC) and the GDP-mannose dehydrogenase (GMD) (algD). These four enzyme activities are necessary for the synthesis of GDP-mannuronic acid, which is the activated sugar precursor for alginate polymerization. For growth-limiting DOT--lower than 10% of air saturation--the increase in mRNA levels of algA, algC and algD with oxygen concentration was accompanied by a strong increase in the activity of the encoded enzymes and the consequent increase in alginate synthesis. However, and despite the upregulation of alginate gene transcription by DOT above 10% of air saturation, the activities of the encoded enzymes either maintained (GMP and GMD) or decreased (PMI and PMM) their levels at high oxygen tensions, leading to a slight decrease in alginate synthesis. This has previously been attributed to the oxidative inactivation of alginate enzymes, particularly of PMM and PMI activities. PMID- 9404504 TI - Analysis of heterogeneity of Corynebacterium diphtheriae toxin gene, tox, and its regulatory element, dtxR, by direct sequencing. AB - The largest diphtheria outbreak in the developed world since the 1960s is in progress in the Russian Federation. Seventy-two Corynebacterium diphtheriae strains from throughout Russia and the Ukraine, selected for temporal and geographic diversity, and 6 reference and control strains were assayed by DNA direct sequencing, and DNA sequences of their diphtheria toxin gene, tox, and the regulatory dtxR gene, were compared to those of the Park-Williams no. 8 strain (PW8). Twenty-eight C. diphtheriae strains had entire tox sequences identical to that of the PW8 strain. Among the remaining 40 strains which were toxigenic, 4 point mutations were detected in the tox gene, one within the A and three within the B subunit gene. All four were silent mutations, indicating that diphtheria toxin is highly conserved at the amino acid sequence level; therefore, changes in the efficacy of the current vaccines would be unlikely to occur. Within the open reading frame of the regulatory dtxR gene, 35 point mutations were detected. Only 15 strains had entire dtxR sequences identical to that of the PW8 strain. Nine amino acid substitutions were found in the carboxyl half of dtxR: 22 and 25 strains differed from the PW8 strain in one and two amino acids, respectively. Given that naturally occurring variations of dtxR might be associated with increased diphtheria toxin production, studies to investigate the association of these point mutations and amino acid substitutions with quantified toxin production in the strains causing the current epidemic are under way. PMID- 9404505 TI - Human uropathogenic and bovine septicaemic Escherichia coli strains carry an identical F17-related adhesin. AB - Pathogenic Escherichia coli produce fimbriae which mediate binding to mucosal cells. Generally, different fimbriae are associated with different tissular tropisms and different host specificities. Genes encoding for pilin and adhesin subunits of two F17-related fimbriae were cloned and sequenced. The first, G fimbriae, are synthesized by a human uropathogenic E. coli strain, and the second, 20K fimbriae, by a bovine septicaemic E. coli strain. We showed that both fimbriae are identical and present a high homology with F17a and F17b fimbriae synthesized by bovine enterotoxigenic E. coli strains. Furthermore, data showed that the G adhesin did not mediate adhesion to human uroepithelial cells, suggesting that it is not responsible for the urinary tropism of the strain and confirming the intestinal tropism specificity of F17-related adhesins. PMID- 9404506 TI - A new fimbrial putative colonization factor (PCF02) in human enterotoxigenic Escherichia coli isolated in Brazil. PMID- 9404507 TI - Detection of Legionella pneumophila in wastewater by nested polymerase chain reaction. AB - The study of Legionella in treated wastewater acquires special importance when this water is used in irrigation by spray, as Legionella is transmitted via the inhalation of aerosols and may consequently represent a health risk. In this study, we applied polymerase chain reaction (PCR) amplification as an alternative method to plate culture for detecting L. pneumophila in twelve heavily biocontaminated samples from a wastewater treatment plant. Moreover, we studied the efficiency of rapid gel filtration methods and filtration through chelating ion exchange resin in the elimination of PCR inhibitors from wastewater samples. When Legionella was investigated by PCR without any previous treatment, no amplification occurred, and when we used chromatographic methods to eliminate PCR inhibitors, nine out of twelve samples became positive. These results indicate the abundant presence of Legionella in wastewater, and although the methods used to eliminate PCR inhibitors are effective in the preparation of clean samples, the possible presence of different metal-organic matter compounds, which are not eliminated, may produce false-negative results. PMID- 9404508 TI - Sequence of DNA 16S/23S spacer region of Leuconostoc oenos (Oenococcus oeni): application to strain differentiation. AB - Leuconostoc oenos is involved in malolactic fermentation occurring during wine making. An increasing number of wines are being inoculated with malolactic starters to control the process, and the identification and differentiation of selected strains are now indispensable both for quality control of production and for commercial purposes. In the present work we evaluated the potential use of the intergenic regions of three L. oenos strains for their differentiation. The three 16S/23S rRNA intergenic spacers were amplified in vitro by PCR, and sequences were compared. The spacer sequence was highly conserved in all strains. Inside this spacer, a tRNA-Ala gene containing an 18-bp sequence stretch which is conserved in all tRNA genes was discovered. This sequence, together with random primers, was used for characterization of ten L. oenos strains by PCR. PMID- 9404509 TI - Electrochemical measurement of trace concentrations of biological hydrogen produced by Enterobacteriaceae. AB - Accurate measurements of the hydrogen gas produced by Escherichia coli and Hafnia alvei pure cultures during glucose metabolism were performed under different growth conditions: stagnant, with magnetic stirring or with vibrational shaking. These measurements were carried out using an electrochemical hydrogen sensor based on a platinum-coated solid polymer electrolyte membrane (Pt-SPE). The results obtained were dependent on the hydrodynamic conditions of the growth, with greater hydrogen production being associated with the stagnant conditions. These measurements will eventually enable us to elucidate whether the pathway used for glucose metabolism is either strictly or mainly anaerobic and to modify experimental conditions so as to influence the reaction. PMID- 9404510 TI - Plant regeneration of indica rice (Oryza sativa) cultivars from mature embryo derived calli. AB - Plant regeneration from seven-week-old callus cultures derived from mature embryos of several indica rice cultivars was achieved with frequencies of morphogenic calli from 10 to 47%. Three media were tested both for callogenesis and plant regeneration. For 3 of the 7 genotypes examined, the best combination of media for plant regeneration was Murashige & Skoog basal medium: MSC (callogenesis) and MSR (regeneration). The rates of callogenesis were not related to the capacity for plant regeneration. Two genotypes CR-1113 and CR-5272 produced the highest number of regenerated green plants. The results of this study suggest that genetic differences could be directly linked to the ability to regenerate in these plant cultivars. PMID- 9404511 TI - Responses of some Nigerian vegetables of plant growth regulator treatments. AB - The effects of single and combined growth regulator treatments of indole-3-acetic acid (IAA), gibberellic acid (GA3) and coconut milk on plant height, yield, chlorophyll and vitamin contents of Abelmoschus esculetus L and Solanum gilo L were investigated. The single growth regulator treatments consisted of 50mg/L, 100 mg/L of IAA and GA3 and 10%, 15% of coconut milk. In case of combined growth regulator treatments, the treatments were 100mg/L IAA + 100mg/L GA3, 100mg/L IAA + 15% coconut milk and 100mg/L GA3 + 15% coconut milk. Control vegetable plants were sprayed with water. Single treatments of 100mg/L IAA,100mg/L GA3. 10% and 15% coconut milk resulted in significantly increased plant height, chlorophyll contents and yield of A. esculentus, H. sabdariffa and S. gilo while only combined treatments of 100mg/L IAA + 10% coconut milk and 100mg/L GA3 + 15% coconut milk had such an effect on A. esculentus and S. gilo but not on H. sabdariffa. Moreover, singletreatments of 100mg/L GA3 and 15% coconut milk caused significantly higher vitamins A, B6 and C contents of treated plants whereas the combined treatments produced such an effect on only vitamin C contents of treated plants. Growth regulator treatments of 100mg/L GA3 and 15% coconut milk were consistently the best out of the entire growth regulator treatments tried with the treated plants having the greatest plant height, yield, chlorophyll and vitamin C contents. PMID- 9404512 TI - [Essential oils of Drimys granadensis (Interaceae) leaves and green fruits]. AB - The composition of the essential oils of the leaves and fruits of Drimys granadensis L. f. (Winteraceae) obtained by hydrodistillation were investigated by GC-MS. The main constituents of the leaves oil were the monoterpenoids 4 terpineol (21.9%), sabinene (16.6%), gamma-terpinene (8.3%) and alpha-terpinene (5.5%), together with the sesquiterpene germacrene-D (10.2%). The main constituents of the unripe-fruit oil were the sesquiterpenoids germacrene-D (23.4%) and drimenol (10.0%). PMID- 9404513 TI - [Essential oil components in leaves and stems of Piper bisasperatum (Piperaceae)]. AB - The composition of the essential oils of the leaves and spikes of Piper bisasperatum Trel. (Piperaceae) obtained by hydrodistillation were investigated by GC-MS. The main constituents of the leaves oil were the sesquiterpenes germacrene D (30.7%), beta-caryophyllene (9.5%) and gamma-elemene (5.9%), together with the monoterpene beta-pinene (5.5%). The main constituents of the spikes (flowers and fruits) were the monoterpenes beta-pinene (17.5%), alpha pinene (14.0%) and the sesquiterpenes germacrene D (19.5%), beta-caryophyllene (7.9%) and ylangene (7.6%). The C6-C1 compounds benzyl benzoate (0.3%) and benzaldehyde (< 0.1%) were identified for the first time in the genus Piper. PMID- 9404514 TI - [Effects of acute and subacute administration of Pimenta dioica (Myrtaceae) extracts on normal and hypertensive albino rats]. AB - The intraperitoneal administration of different extracts of Pimenta dioica (L.) Merrill (Myrtaceae) to conscious normotensive and hypertensive rats caused a depression of the central nervous system (CNS). The intensity of this depression depends on the dose. Analgesic and hypothermic effects were also observed. The total aqueous extract was more effective than the ethanolic extract and the final aqueous fraction was the most effective. The peritoneal irritation caused by the extract explains only partially the depressive effect over the CNS. When the final aqueous fraction was given orally to SDN and SHR rats during 14 days there was no observed change on the sistolic blood pressure, heart rate and weight of the animals. PMID- 9404515 TI - [Pharmacologic activity of the aqueous wood extract from Quassia amara (Simarubaceae) on albino rats and mice]. AB - All the assays were done with an aqueous preparation of dry wood from Quassia amara (Simarubaceae). For the hippocratic assay, 12 female SDN rats were used, with an average weight of 144 g and separated in three groups of four individuals each. The dose used were 500 mg/kg and 1,000 mg/kg and the control group received 0.5 ml of distilled water. The extract administration and the observation of the animals were done daily during nine days. Acute toxicity of the preparation was studied with 25 male NGP mice with an average weight of 20.13 g, in groups of five individuals per dose. The oral administration was carry out with the following doses: 250, 500, 750 and 1,000 mg/kg, the control group received 0.5 ml of distilled water. No sign of acute toxicity was observed at any dose. For the toxicity analysis via intraperitoneal injection 15 male NGP mice were assigned to five groups (5 animals each) with doses of 500 and 1,000 mg/kg and a control group with 0.5 ml of distilled water. The group with the dose of 500 mg/kg, presented acute toxicity signs with a 24 hr recovery, and the 1,000 mg/kg dose was lethal to a 100% within 24 hr. The measuring of the peristaltic activity (movement of the intestinal content) were performed on 30 NGP male mice with an average weight of 22 g assigned to three groups of ten individuals each. One dose of 500 mg/kg and 1,000 mg/kg were orally administrated to each experimental group and 0.5 ml of distilled water to the control group. The marker used was activated carbon, orally supplied to every mice 30 min after the administration of the aqueous extract. The animals are decapitated and the measurement of the carbon motion in the small intestine was done after 30 min. Both dose increased the intestinal movement compared to the control group, but only the 1,000 mg/kg dose showed a statistically significant difference (p < or = 0.05). PMID- 9404516 TI - Experimental Trypanosoma rangeli infection in a murine model. AB - Trypanosoma rangeli experimental murine infections were performed in order to study parasitemias and anti-parasite antibody levels. Three groups of mice were used: a) mice infected with metatrypomastigotes derived from infected bugs; b) mice which received four reinoculations of metatrypomastigotes and c) mice immunosuppressed with cyclophosphamide. The results showed that bloodstream parasites can be found from the first day post inoculation reaching a peak at day 5 or 7 and then start to decline. Parasites disappeared completely from the circulation after 20-25 days. However in the immunosuppressed group, parasites were found in blood up to 45 days post infection. The humoral immune response was monitored using an ELISA test and low levels of specific IgG and IgM immunoglobulins were found. However the IgG titers were lower than the IgM. One could conclude that IgM was the predominant immunoglobulin isotype induced in a T. rangeli experimental infection because the highest titers were observed in the reinoculated group. IgM antibodies also showed the most prominent crossreactivities with T. cruzi antigens. PMID- 9404517 TI - Toxoplasma gondii (Eucoccidia: Sarcoystidae) dissemination pattern in rats after oral infection with oocysts of an avirulent strain. AB - Differences in Toxoplasma gondii dissemination in white rats (Sprague Dowley) and mice (Wistar) after oral oocyst inoculation are described. Groups of five animals (both hosts) were infected per os with oocysts of the TCR-2 avirulent strain and the dissemination pattern was compared in brain tissue or by serology. Early dissemination was similar in both species. One hr after infection the parasite was present in blood and peritoneal exudate as well as in heart, lung, liver, spleen, lymph nodes and brain. However, after five days there were important differences between both hosts and after 30 days, the parasite was detected only in rat heart and brain, while in mice it persisted in fluids and all organs. PMID- 9404518 TI - [Host plants of Bemisia tabaci (Homoptera: Aleyrodidae) in Cuba]. AB - The sweet potato white fly, Bemisia tabaci, is an important pest of tomatoes and beans, among other crops, which transmits viral diseases. Since the second quarter of 1989 a significant population increase of this pest has been noted in several cultivated plants. From 1989 to 1992, a survey was done throughout the country, chiefly in vegetable and bean-producing areas. They occur in 119 species (42 families), a great increase over the previous record of four species. Worldwide, this report represents 50 species and six families which are new records. PMID- 9404519 TI - [Host plants of Cerambycidae (Coleoptera) from northwest and central Argentina]. AB - Seventy four species of Cerambycidae (Coleoptera) were reared from forty eight host plants found into Yungas, Chaco and Prepuna phytogeografical provinces. The host plants belong Anacardiaceae, Asclepiadaceae, Bignoniaceae, Cactaceae, Cucurbitaceae, Euphorbiaceae, Juglandaceae, Leguminosae sensu lato (Caesalpinaceae, Fabaceae, Mimosaceae), Loranthaceae, Rhamnaceae, Rutaceae, Salicaceae, Sapindaceae, Solanaceae, Ulmaceae and Vitaceae. Some species of Cerambycidae and their host plants show similar distribution patterns: from northeastern Brazil through Paraguay and reach the Argentina at both sides of the Chaco Provine: at the east, along the river systems through Buenos Aires, and the at the west into forests of the Yungas Province or into North Sierra Chaco, a probably relictual community. Others species are restricted to the South America area of Prosopis, that comprise the Chaco, Monte and Espinal biogeographical provinces, and live in host plants of chaquenian lineage mainly Leguminosae sensu lato and Ulmaceae. PMID- 9404520 TI - [Host plants of Cerambycidae (Coleoptera) species in Espinal and Buenos Aires Province, Argentina]. AB - The host plants of Cerambycidae (Coleoptera) in the Province of Espinal (Chaquenian Dominion) and the province of Buenos Aires are mainly exotics, cultivated or naturalized, the last ones growing spontaneously and forming little forests. Forty two species of Cerambycidae are reported using these plants and other native species as hosts. Hylotrupes bajulus, originally introduced and a serious pest of furniture, is reported for first time in spontaneous pine forests in Sierra de la Ventana, south of the province of Buenos Aires. PMID- 9404521 TI - [Host plants of Cerambycidae (Coleoptera) in the northeast of Argentina]. AB - New host plants and localities are recorded for 84 species of Cerambycidae of Chaco Province (Chaquenian Dominion) and Paranaense Province (Amazonian Dominion) in northeastern Argentina and Brazil. Host plants belong to Anacardiaceae, Asclepiadaceae, Apocynaceae, Bignoniaceae, Bombacaceae, Cactaceae, Caesalpinaceae, Capparidaceae, Casuarinaceae, Fabaceae, Mimosaceae, Moraceae, Nyctaginaceae, Polygonaceae, Rhamnaceae, Rutaceae, Sapindaceae, Sapotaceae and Ulmaceae. First records for Argentina and host plants: Compsocerus barbicornis Serville 1834, Desmiphora lenkoi (Lane 1959), Neocompsa serrana (Martins 1962) and Trachysomus dromedarius (Voet 1778). First host plants records of rare or uncommon Argentine species of Cerambycidae are Methia tubuliventris Gounelle 1913, Paraleptidea femorata Gounelle 1913 and Oncideres pepotinga Martins 1981. PMID- 9404522 TI - [Population parameters of three parasitoids (Hymenoptera: Scelionidae, Encyrtidae) hosted by the predator Podisus nigrispinus (Heteroptera: Pentatomidae)]. AB - The reproductive parameters of Telenomus podisi, Trissolcus brochymenae (Hymenoptera: Scelionidae) and Ooencyrtus sp. (Hymenoptera: Encyrtidae) were studied for three generations on their natural host Podisus nigrispinus (Dallas, 1851) (Hemiptera: Pentatomidae). A parasitism rate of 10 eggs/day/species was used under laboratory conditions of 27 +/- 1 degrees C, 60 +/- 10% RH and 14 h of photophase. No significant differences were detected between generations in terms of net reproductive rate (Ro), intrinsic rate of natural increase (rm), generation time (T) and the effect of doubling time on population increase (DT). However, T. podisi and T. brochymenae had significantly higher Ro and rm values, and lower T and DT values, with rm 1.59 and 1.50 times higher than Ooencyrtus sp., respectively. In addition the former species also had shorter egg-adult period, greater longevity of females and reproduction period than the other species. These results confirm the greater reproductive potential of T. podisi and T. brochymenae, which are commonly found parasitizing P. nigrispinus. PMID- 9404523 TI - Effect of Eucalyptus feeding in the development, survival and reproduction of Tynacantha marginata (Heteroptera: Pentatomidae). AB - The effect of feeding on Eucalyptus leaves of Tynacantha marginata (Heteroptera: Pentatomidae) was studied in Brazil. The use of this plant as a complementary food source for the predatory bug increased its oviposition period, generation time, and net reproductive rate. Eucalyptus leaves apparently had no influence on the survival and development of T. marginata. PMID- 9404524 TI - [Seasonal variation of three Culex species (Diptera: Culicidae) and its parasites and pathogens in Punta Lara, Buenos Aires, Argentina]. AB - The seasonality of immatures, imagoes and pathogens of Culex dolosus Lynch Arribalzaga, Culex intrincatus Brethes and Culex maxi Dyar from Punta Lara (Buenos Aires province, Argentina) was monitored weekly (larvae and pupae) or fortnightly (adults), from 1989 to 1991. Culex dolosus was present during all months. The larvae were parasitized by Geotrichum candidum Link ex Person, Smittium morbosum var. rioplatensis Lopez Lastra, Coelomomyces sp., Achlya sp, Amblyospora dolosi Garcia & Becnel and Strelkovimermis spiculatus Poinar & Camino. Conidia of Hyphomycetes, filarial worms of Onchocercidae and S. spiculatus were detected in adults. Cx. intrincatus was present from January to May; the immatures were parasitized by G. candidum, S. morbosum, Achlya sp. and Amblyospora sp. Larvae of Cx. maxi were collected from January to April, and adults in all months except for June and July. S. morbosum was the only parasite found in this mosquito. PMID- 9404525 TI - [New therapeutic principle in the management of hypertension]. PMID- 9404526 TI - [Signal-enhanced Doppler examination. Better diagnosis with a new contrast medium for ultrasonography]. PMID- 9404527 TI - [Effective cholesterol and triglyceride lowering with atorvastatin]. PMID- 9404528 TI - [Tolperisone in spasticity and muscle spasm. Tolperisone in multiple sclerosis]. PMID- 9404529 TI - [Epilepsy therapy: lamotrigine--retrospective and prospective study]. PMID- 9404530 TI - [Depression in advanced age. Response and tolerance speak for SSRI]. PMID- 9404531 TI - [Therapy of Parkinson disease. Continuous physiologic dopaminergic stimulation with carbergoline]. PMID- 9404532 TI - [Advances in depression therapy. Sertralin: highly selective SSRI with low interaction potential]. PMID- 9404533 TI - [Depression therapy becoming easier]. PMID- 9404534 TI - [Lamotrigin--an effective partner in the management of epilepsy]. PMID- 9404535 TI - [COMT inhibition with tolcapone]. PMID- 9404536 TI - [10th Psychiatric World Congress. Madrid, August 1996. Schizophrenia: how to improve compliance]. PMID- 9404537 TI - Seasonal dynamics of the tissue levels of total protein, free amino acid and ribonucleic acid in an Indian air-breathing teleost, Channa punctatus. AB - The tropical freshwater air-breathing teleost, Channa punctatus, reveals a seasonal metabolic reorganization with reference to its biochemical tissue compositions. The summer-adapted and winter-adapted C. punctatus seem to be two biochemically distinct populations. Besides, the different reproductive phases viz. prespawning, spawning, post-spawning, preparatory-I and preparatory-II are also biochemically distinct. PMID- 9404538 TI - A comparison of blood composition in the common carp (Cyprinus carpio L.) using samples obtained from caudal vessels and dorsal aorta cannulae. AB - Blood samples have been taken from chronic cannulae in the dorsal aorta of Carp under anaesthesia and at intervals of 24, 29 and 101 hours later. A few minutes after the 29 hours and 101 hours sampling was completed similar quantities (1 ml) were withdrawn from a caudal vessel. All samples were treated in the same way and determinations made of haematocrit value, total protein, glucose concentration and activity of the following plasma enzymes-acetylcholinesterase (AChE), glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT) and lactic dehydrogenase (LDH). Comparisons between dorsal aorta and caudal samples at 29 hours and 101 hours showed no significant differences for any of the enzyme activities that were tested. However significant (paired "t"-test, P < 0.05) differences were observed for haematocrit and glucose concentration. It is suggested that the higher haematocrit and lower glucose of caudal samples are consistent with the expected differences between arterial and venous blood. Apart from these differences which reflect normal physiological function, values obtained for samples from the two sites were identical. It is concluded that blood sampling from caudal vessels is a satisfactory method to obtain blood for biomonitoring purposes at least for those enzymes commonly used in recent surveys. PMID- 9404539 TI - Study of the effects of brief exposure to an organophosphorous insecticide (methidathion) on blood characteristics of carp (Cyprinus carpio). AB - 1. Carp with the dorsal aorta cannulated have been used to study the effects of a 5 hour exposure to methidathion at concentrations of 2 or 6 mg/litre. Blood samples taken during control, exposed and recovery periods were used to determine plasma acetylcholinesterase (AChE) activity, haematocrit value, mean cell volume, total plasma protein, and filtration time through Nucleopore filters containing pores of 8 microns. 2. A drastic inhibition of AChE activity was observed and this continued during the recovery for at least 5 days. Changes in other blood parameters were less marked and recovered soon after fish were returned to non polluted water. The increased haematocrit and decrease in filtration time suggests some impairment of gas transfer during exposure but the depletion of AChE activity and associated muscular and neural disturbances are more serious results of pollution with this organophosphate insecticide. PMID- 9404540 TI - In vivo induction of sister chromatid exchanges (SCE) in a tropical fish, Etroplus suratensis (Bloch). AB - Sister chromatid exchange (SCE) test was applied to a tropical cultured fish, Etroplus suratensis by exposing to known mutagens. Intramuscular injections of methyl methane sulphonate (MMS) and cyclophosphamide (CP) for an exposure period of 96 h resulted in significant increase in the frequency of SCE in gill tissues. MMS induced dose dependent increase in SCE. The findings revealed that the test species in the present study may be used in mutagenicity assays for screening environmental pollutants. PMID- 9404541 TI - Ultrastructural pathology of degenerating "dark" granule cells in the hippocampal dentate gyrus of adrenalectomized rats. AB - Adrenalectomy-evoked delayed degeneration and death of granule cells in the hippocampal dentate gyrus (DG) of the rat brain were studied by means of electron microscopy and a recently elaborated silver method that selectively stains the "dark", collapsed neurons in a Golgi-like manner. At the light microscopic level, the silver technique revealed degenerating granule cells located exclusively in the dentate gyrus; other glucocorticoid receptor-containing regions of the brain were not affected. The silver-stained cell bodies were shrunken, most of the dendrites had a beaded appearance, and the stained axons could be traced along their route to the CA3 pyramidal neurons of the hippocampus. The analysis of 2.5 microns thick Epon-embedded sections stained with toluidine blue revealed hyperchromatic, dark granule neurons and their remains and a heavy glial activity in the vicinity of collapsing neuronal profiles. At the ultrastructural level, early and late stages of neuronal degeneration were observed. The early phase was characterized by markedly increased electron density, a massive shrinkage of the whole somato-dendritic domain, vacuolization of mitochondria, swelling of the nucleolus and condensation of the nuclear chromatin. In the late stage, subcellular organelles were hardly recognizable due to the extremely high electron density and dramatic shrinkage of the cytoplasm. These profiles exhibited disintegration of the cellular organelles and loss of their afferents. Concomitantly, disintegration of granule cell dendrites (clasmatodendrosis) and lifting of "dark" mossy fibers from cell bodies and dendrites of CA3 pyramidal neurons were observed. In the latter cells, this partial denervation caused no apparent signs of ultrastructural alterations. Proliferation of astrocytes and microglial cells was also obvious as they engulfed and eliminated the degenerating neuronal elements. Degenerating neurons frequently occurred adjacent neurons with normal morphology. These morphological features indicate that the delayed degeneration of hippocampal granule cells following adrenalectomy might proceed through a cytoskeletal collapse terminating in cell death. PMID- 9404542 TI - Substance P immunoreactive elements in the nervous system of earthworm (Lumbricus terrestris). AB - Substance P immunoreactive elements were detected in the nervous system of the earthworm, Lumbricus terrestris using peroxidase-antiperoxidase method. Each part of the central nervous system contains immunoreactive perikarya. The central neuropil shows no stained fibers. Immunopositive nerve cells were found in the stomatogastric ganglia as well as nerve cells and fibers in the wall of the alimentary canal. In the body wall, epidermal sensory cells and fibers surrounding the chaetae display immunoreactivity. The results are compared with those described previously by other authors. PMID- 9404543 TI - Chromatographic screening of growth inhibitors for Cladosporium herbarum in the extracts of calcareous red algae. AB - The thin-layer chromatographic screening of growth inhibitors for an imperfect fungus was carried out in two types of calcareous red algae, Corallina pilulifera (Corallinaceae) and Lithothamnium pacificum (Corallinaceae) and Crathromorchum circumscriptum (Corallinaceae). Several substances having positive activities on Cladosporium herbarum were detected in the alcoholic extracts of the marine plants. The observed differences among the substances can be attributed to the morphological characteristics of the sea weeds. PMID- 9404544 TI - Factors affecting transient expression of vector constructs in wheat protoplasts. AB - Direct uptake of reporter gene constructs with the bacterial beta-glucuronidase (GUS) gene fused to various promoters was achieved to embryogenic cell suspension culture-derived protoplasts of GK Sagvari winter wheat (Triticum aestivum L.) with polyethylene glycol (PEG) treatment. Based on GUS specific activity values, it was found that Mg2+ with PEG at 20% final concentration can significantly increase the transient expression in wheat protoplasts in comparison to the Ca(2+)-containing medium. The optimum incubation time in transformation mixture was 5-10 min at 25 degrees C. Transient GUS expression as detected by spectrofluorimetry was positively correlated with the time elapsed after DNA uptake (with maximum activity at 48 h), and the incubation time in GUS reaction mixture. It was also found that the protoplast culture medium plays an important role in the efficiency, and the treated wheat protoplasts cultured in KM medium showed a higher GUS activity than those kept in GM medium. Among the five plasmid constructs 6-16-fold higher promoter activity has been achieved with pKM794 driven by CaMV 35S promoter plus two enhancer elements than with the other constructs tested. PMID- 9404545 TI - Effects of dietary L-carnitine supplementation and protein level on performance and degree of meatness and fatness of broilers. AB - Feeding high-protein diets to broilers is a known means for improving performance and carcass composition and quality, however, the combined effects of dietary protein (CP) level concurrent with L-carnitine (CN) supplementation are not known. Performance and carcass traits of broilers fed CN-supplemented diets of different CP-levels were studied from 18 to 53 days of age. Three isocaloric diets containing 18, 20 or 22% CP were formulated, with or without added CN (50 mg/kg), and used. Apart from CP level, supplemental CN attained significantly higher body weight gain (BWG) and feed conversion (FC), means of BWG were 351 vs. 332 and 371 vs. 353 g, and those of FC were 1.25 vs. 1.33 and 1.73 vs. 1.84 for the first and second week of the experimental period, respectively. Amount and percentage of abdominal fat and ether extract (EE) contents of breast meat of 53 day-old broilers were significantly reduced (means of AF were 43.5 vs. 51.5 g, equivalent to 1.98 vs. 2.39%, those of EE were 1.82 vs. 2.24%) in response to CN supplementation. A significant interaction between added CN and CP level was noted on BWG and FC of broilers from 18 to 32 days of age. PMID- 9404546 TI - Influence of dietary L-carnitine on performance and carcass quality of broiler chickens. AB - Effects of dietary L-carnitine supplementation on the performance, carcass yield and components, abdominal fat and composition of meat of broiler chickens were investigated. Ross broiler chicks, 18-day-old were used in this experiment. Four experimental diets were formulated, by adding three levels of supplemental L carnitine (50, 100 or 150 mg/kg) to a basal diet and used from 18 to 46 days of age. Apart from the level of supplementation, L-carnitine (CN) addition resulted in significant increases (P < 0.05) in body weight gains (BWG) of broilers (means were 397 vs. 365 and 410 vs. 377 g, for the first and second weeks of the experimental period, respectively). Abdominal fat (AF) contents of 46-day-old broilers were significantly decreased, both as absolute weights (P < 0.05) and as percentages of body weight (P < 0.01), by added CN (means of AF were 36.8 vs. 44.8 g, equivalent to 1.87 vs. 2.32%). Level of added CN had no effect on BWG or AF of broilers. It was concluded that the effectiveness of supplemental L carnitine for improving BWG and/or decreasing AF of broilers may depend on the age at which L-carnitine is added. Under the conditions of this study, a supplementation of 50 mg L-carnitine per kg diet proved to be effective. PMID- 9404547 TI - Simultaneous isolation of both RNA and DNA from many small tissue samples. PMID- 9404548 TI - A comparison of the clinical performance, contraceptive efficacy, reversibility and acceptability of Norplant implants and Ortho Gynae T380 intrauterine copper contraceptive device. AB - In this comparative study, the five-year continuity rate of 53.7% in the Norplant implants group was comparable to that of 52.7% in the copper IUD group. The difference was not statistically significant. Only one accidental pregnancy occurred during the five years of copper IUD use. Desire for future pregnancy was the main reason for removal in the Norplant implants group (35.9%) while expulsion of the IUD (13.2%) was the main reason for removal of the copper IUD. Menstrual disturbance was not a major side-effect in either group. The post removal conception rates of 78.6% in the Norplant implants group and 75.0% in the copper IUD were good and comparable. Both the Norplant implants and copper IUD are acceptable and effective contraceptive methods in Singapore. PMID- 9404549 TI - The frequency and spectrum of congenital anomalies in natural family planning users in South America: no increase in a case-control study. NFP-ECLAMC Group. Natural Family Planning. Latin-American Collaborative Study of Congenital Malformations. AB - Users of natural family planning (NFP) practice periodic abstinence, leading many to reason that such couples should show increased anomalies in offspring as a result of fertilization involving aging gametes. In an effort to complement our NFP cohort study, we currently conducted a case-control study in the same region (South America) in which the largest number of cases have been recruited for our cohort NFP study. During 1992-94, 5324 case-control pairs of mothers were interviewed during the immediate postpartum period in 18 maternity hospitals participating in the Latin-American Collaborative Study of Congenital Malformations: ECLAMC (Spanish acronym for Latin-American Collaborative Study of Congenital Malformations). Natural family planning (NFP) usage was recorded in 6% of mothers in the ECLAMC sample studied (n = 10,648). Overall, no significant differences in frequency of NFP usage were observed between malformed cases (349/5324 = 6.6%) and normal controls (303/5324 = 5.7%) (chi 2 = 3.3; df = 1; p > 0.05). No significant differences in sex ratios were observed between children of NFP user and non-user mothers. Of special interest is the lack of association between NFP and Down syndrome, the sentinel phenotype for the hypothesis of delayed fertilization (aging gametes). PMID- 9404550 TI - Current status of injectable hormonal contraception, with special reference to the monthly method. AB - Injectable contraceptives are a valid option in every family planning program. Contraceptives which are administered every 2 or 3 months, containing only progestogen agents (DepoProvera, Noristerat) have proven efficacious and do not show long-term safety problems. They differ from other contraceptives in their long lasting action and by not presenting the contraindications of the estrogens. Their most prominent side-effect is the irregularity of cyclic bleeding. Although bleeding irregularities are not life threatening, many users stop the treatment for that reason. Monthly contraceptives comprising progestogens and estrogens, maintain or improve the high efficacy of the earlier forms and have the added benefit of allowing bleeding to resemble the physiologic one. This increases acceptability and the continuation rate. There is no long-term inconvenience. At this point, the greatest experience is with the formulation known as Topasel or Perlutal. Other formulations (Cyclofem, Mesigyna) are beginning to be commercialized and their characteristics must still be confirmed through daily use. Indications, contraindications, precautions and warnings for the use of monthly injectable contraceptives are basically identical to those of the combined oral contraceptives, as are the side-effects. Efficacy, though, proves to be superior, which can be correlated to a simpler method of use and less risk of error when using it. Main motivation factors are: efficacy, simplicity in usage, reversibility and confidentiality. PMID- 9404551 TI - Treatment of bleeding problems associated with progestin-only contraceptives: survey results. AB - Simple, well-documented, effective and inexpensive management approaches are needed for the short-term control of bleeding in progestin-only contraceptive (POC) users. Long-term continuation may improve if acceptable approaches are found to ameliorate these short-term bleeding problems. Family planning providers and researchers were surveyed on the treatment regimens they used for POC associated bleeding disturbances. Results from this limited survey indicate that no clear indication exists regarding the duration and, severity of bleeding and spotting requiring pharmacologic intervention. Counseling was seen as a critical factor when considering treatment for these problems. This report presents the results of this survey on the different treatment regimens being used by providers throughout the world to treat POC-associated bleeding disturbances. PMID- 9404552 TI - Factors associated with unintended pregnancy. AB - This research was designed to identify determinants of unintended pregnancy among women attending family practice or family planning clinics. Survey data were collected from 95 women who were categorized according to whether or not they had experienced an unintended pregnancy. Women reporting unintended pregnancy were younger, reported earlier sexual debut and a greater number of sexual partners than those not having experienced an unintended pregnancy. Those who had avoided unintended pregnancy displayed higher levels of preventive sexual self-efficacy, had more confidence in their ability to use contraceptive methods, perceived more negative consequences associated with having children in the near future, and believed pregnancy among unmarried women to be less acceptable than did women who had had unintended pregnancies. PMID- 9404553 TI - Safety, functionality and acceptability of a prototype polyurethane condom. AB - Male condoms made from synthetic materials offer an alternative to latex condoms that may be more acceptable to users, thereby potentially resulting in more protected acts of intercourse. A prospective, noncomparative clinical study was conducted to evaluate the safety of using certain polyurethane materials to make condoms. Fifty-one healthy, contracepting, mutually monogamous couples were recruited between June 30 and November 24, 1993 to use a prototype roll-on polyurethane condom developed by Family Health International. Couples were to use the condoms for 10 consecutive acts of vaginal intercourse over a 4-week period. Baseline and postexposure genital examinations, including colposcopy for female participants, were performed. Fifty couples completed the study requirements and 517 acts of intercourse occurred using the condoms. Two adverse events were reported: irritation of introitus in a female participant and a small irritated erythematous lesion on a male participant's penis. Neither event was considered to be serious and both were resolved without treatment. Breakage and slippage rates were similar to those reported for latex condoms. These results suggest that polyurethane condoms represent a safe, functional and acceptable alternative to latex condoms. PMID- 9404554 TI - Challenges in epidemiologic allergy research. PMID- 9404555 TI - Citations and journal impact factors: questionable indicators of research quality. PMID- 9404556 TI - Reduction of latex-allergen content in Swedish medical catheter balloons--a survey of 3 years' production. AB - Anaphylactic reactions after intravascular exposure to natural rubber latex (NRL) have been reported. Thus, there is an urgent need to produce medical devices with the lowest possible latex-allergen content. The latex-allergen concentration in extracts prepared from 92 lots of medical catheter (MC) balloons, manufactured by Nolato Polymer AB, Torekov, Sweden, from April 1993 to March 1996, was measured with an EAI (IgE antibody inhibition) assay. Inhibitory capacity was expressed in arbitrary units/ml (U/ml) in relation to reference NRL sap, given an arbitrary value of 1000 U. Extracts from randomly selected lots were measured for protein by the modified Lowry method. Water leaching, chlorination, and treatment with savinase were used experimentally to study reduction of the latex-allergen content. The latex-allergen content in extract from the regular MC balloons varied from 0.1 to 2.9 U/ml. All the methods used to reduce the allergen content were effective, and increased leaching stabilized the allergen content at a low level. The protein concentration of the extracts varied between 9 and 100 mg/l. No correlation was found between protein and allergen content. As a result of this study, the manufacturer has extended the stage of water leaching in the production process. This study shows that cooperation between immunologists and manufacturers may result in product development and improvement. PMID- 9404557 TI - In vivo relevance of CD30 in atopic dermatitis. AB - CD30 expression was evaluated by immunohistochemistry in lesional skin biopsies of eight patients with active atopic dermatitis (AD) and three patients with allergic contact (nickel-induced) dermatitis (ACD). CD30 expression was also assessed in a large panel of CD4+ and CD8+ T-cell clones generated from the skin biopsies of four patients with AD. Finally, the levels of soluble CD30 (sCD30) were measured in the serum of 41 patients with AD, 19 patients with ACD, and 60 healthy controls. In all specimens of lesional AD skin, where the great majority of infiltrating cells were CD4+ T cells, remarkable numbers of cells were CD30+, whereas virtually no CD30+ cells were found in the skin of patients with ACD. In CD4+ T-cell clones generated from the lesional AD skin, most of which produced both interleukin (IL)-4 and interferon-gamma (IFN-gamma) (Th0-like cells) or IL-4 and IL-5, but not IFN-gamma (Th2-like cells), CD30 expression directly correlated with the ability to produce IL-4 and IL-5, but was inversely related to IFN-gamma production. High levels of sCD30 (correlated with disease activity: r = 0.618) were detected in the serum of most AD patients, whereas there was no increase of sCD30 levels in the serum of patients with ACD. These data support the view that Th0/Th2-type responses predominate in the skin of patients with AD and suggest that the presence of CD30+ T cells in tissues and/or increased levels of sCD30 in biologic fluids are indicative of Th2-dominated responses. PMID- 9404558 TI - Incidence of asthma in adults--report from the Obstructive Lung Disease in Northern Sweden Study. AB - Incidence studies offer a better opportunity to study risk factors for asthma than do prevalence studies. However, regular prospective follow-ups of large cohorts are difficult to perform, and that is why direct measurement of the incidence rate of asthma is almost impossible. Thus, cross-sectional follow-up studies of defined cohorts can be used to provide data on incidence. In 1986, a postal questionnaire survey on respiratory symptoms and diseases was performed in the northernmost province of Sweden. The population sample comprised all subjects born in 1919-20, 1934-5, and 1949-50 in eight representative areas of the province, which comprises 25% of the total area of Sweden. Completed answers were given by 5698 subjects (86%) of the 6610 subjects invited to the study. In 1992, the cohort was invited to a follow-up survey during the same season as in 1986, and 6215 subjects were traced. Of the 5393 subjects who answered the questionnaire, 4932 had participated in the 1986 survey, or 87% of those who participated in 1986. For the period 1986-92, the cumulative incidences of asthma were 4.9 and 5.0%, respectively, as assessed by the questions, "Have you ever had asthma?" and "Have you been diagnosed as having asthma by a physician?" Thus, the results indicate a mean annual cumulative incidence of asthma of 0.8%. After correction of the results for subjects who were diagnosed as having asthma in the clinical part later in the 1986 study, the mean annual cumulative incidence of asthma was found to be 0.5%. Risk factors were family history of asthma (OR 3.46) and current and former smoking, while female sex was a strong trend. PMID- 9404559 TI - Genes, environments, and sex: factors of importance in atopic diseases in 7-9 year-old Swedish twins. AB - Various atopic manifestations among adults have been shown to be influenced mainly by genetic factors. With the increase in prevalence of atopic diseases in recent years, especially among children, a great deal of attention has been given to environmental causes. In a study of 1480 Swedish twin pairs, 7-9 years old, we examined the importance of genetic and environmental factors in asthma, hay fever, eczema, and urticaria. Structural equation model fitting showed 33-76% of the variation in liability to the diseases to be due to genetic effects. Shared environmental effects were also important for hay fever and urticaria in both sexes and for eczema among girls. The clustering of atopic disease in families was almost entirely due to a common set of genes, but each disease manifestation also seemed to have specific genes of importance. Investigation of unlike-sex twins showed that boys had a higher cumulative incidence of asthma and hay fever than girls, whereas girls had a higher incidence of eczema. Thus, it may be concluded that although genetic factors are of major importance in atopic manifestation in children, both environmental and sex-related factors play a role. PMID- 9404560 TI - Monitoring of serologic immune parameters in inflammatory skin diseases. AB - This paper deals with the correlation of clinical scoring and serologic markers of inflammation in atopic dermatitis and psoriasis. Serum eosinophil cationic protein (ECP), soluble interleukin-2 receptor (sIL-2R), total serum IgE, IgG and IgM anti-IgE antibodies, and IgE immune complexes were evaluated in monitoring inflammatory skin diseases such as atopic dermatitis and psoriasis. Well established clinical activity scores were used as standards in recording skin improvement under treatment in a clinical setting. Serum ECP was found to be increased in both atopic dermatitis and psoriasis patients compared to normal controls; sIL-2R and IgE immune complexes were increased only in atopics with increased serum IgE. Anti-IgE antibodies did not show any deviation in both groups of patients. There was a significant elevation of sIL-2R and IgE immune complexes and a nonsignificant elevation of ECP in high-IgE atopics in comparison to those with normal serum IgE. In both groups of patients, there was a significant reduction of ECP and sIL-2R accompanying the improving skin condition. Serum IgE and the other immune parameters failed to respond. In contrast to other studies, serum ECP failed to correspond significantly with disease activity in our study. Our results showed measurable changes of ECP and sIL-2R for atopic dermatitis and/or psoriasis under treatment, but comparison to clinical scores remains difficult due to the different basis of the two systems. The only significant correlation was established for relative changes in sIL-2R and psoriasis area and intensity (PASI), a correlation which might be a useful approach in psoriasis. PMID- 9404561 TI - Effects of rapamycin, cyclosporin A, and dexamethasone on interleukin 5-induced eosinophil degranulation and prolonged survival. AB - Interleukin-5 (IL-5) enhances eosinophil degranulation and prolongs eosinophil survival. Rapamycin, cyclosporin A, and dexamethasone have been shown to influence either cytokine transcription, cytokine-mediated signalling, or degranulation by granulocytes. The study aimed to determine whether these agents inhibited IL-5-enhanced eosinophil survival or degranulation. Peripheral blood eosinophils were isolated from atopic subjects. The effects of serial dilutions (10(-6)-10(-9) M) of these drugs or vehicle control on 1) the viability of eosinophils cultured (1-5 days) in the presence and absence of recombinant human IL-5, as measured by propidium iodide staining and flow cytometry, and 2) degranulation of eosinophils preincubated (45 min) with rhIL-5 or medium control, as measured by eosinophil cationic protein (ECP) release after stimulation with serum-coated Sephadex beads, were assessed. Dexamethasone and rapamycin produced significant, concentration-dependent inhibition of IL-5-enhanced eosinophil survival at pharmacologic concentrations, whereas cyclosporin A did not. Prior incubation of eosinophils with IL-5, as compared with medium control, significantly enhanced ECP release by eosinophils on subsequent exposure to serum coated Sephadex beads. Cyclosporin A and rapamycin significantly inhibited IL-5 enhanced ECP release in a concentration-dependent fashion, whereas dexamethasone did not. All three drugs had no significant effect on eosinophil survival and degranulation in the absence of IL-5. Our results suggest that immunosuppressive drugs may inhibit IL-5-mediated mechanisms in eosinophils which result in enhanced survival and release of granule contents. These findings may be relevant to the further development of therapeutic strategies in allergic diseases. PMID- 9404562 TI - Serum tryptase levels in adverse drug reactions. AB - We evaluated the usefulness of individual tryptase levels and variations after adverse drug reactions in 64 patients. Our aim was to find a tool for the diagnosis of drug allergy. Thirty-seven subjects were confirmed to have drug allergy, 12 had nonsteroidal anti-inflammatory drug (NSAID) reactions, five had negative controlled drug challenges (NAAR), and 10 had symptoms after placebo intake (PLA). Serum tryptase levels greatly increased after anaphylactic shocks (2242%) and anaphylaxis (710.5%). Patients with allergic urticaria and those with idiosyncratic responses to acetylsalicylic acid (ASA) exhibited a small increase in serum tryptase (49.5% and 38.2%, respectively). In the other two groups (NAAR and PLA), no variation in this serum protease was observed. The time of appearance of the serum tryptase peak differed considerably among patients with similar clinical reactions (from 30 min to 6 h) and was independent of the latent period, severity of symptoms, or the amount of tryptase released. We conclude that serum tryptase determinations are helpful in the diagnosis of anaphylactic shock and anaphylaxis, but serial measurements may be needed to confirm mast-cell participation in milder reactions. PMID- 9404563 TI - Comparison of four different measures of bronchial responsiveness in asthmatic children. AB - Although several tests are available to assess the presence and severity of bronchial hyperresponsiveness (BHR), there is no agreement on the most appropriate stimulus. The most commonly used stimuli are methacholine, histamine, and exercise. Daily peak expiratory flow (PEF) variation has been reported to correlate with the severity of BHR, and in recent years this has been widely used because of its noninvasiveness and ease of performance. This study was carried out to determine the relationship among these four commonly used measures of bronchial responsiveness in asthmatic children. For this purpose, 12 asthmatic children of varying disease severity were recruited. Subjects underwent three challenges on 3 separate days in 1 week. During the week preceding the challenges (methacholine, histamine, and exercise), patients recorded PEF three times a day. All patients had PC20 less than 8 mg/ml with methacholine and histamine. Patients with PC20 greater than 3.5 mg/ml for both methacholine or histamine had negative exercise challenges. The strongest correlation was between histamine and methacholine (r = 0.95). Exercise-induced bronchospasm had substantial and significant correlation with the other three measures. No significant correlation was observed between PEF variability and histamine or methacholine. The varying degrees of relationships among the four commonly used measures suggests that each method yields information on different but related phenomena. More than one measure may be required to detect the different aspects of asthmatic bronchial responsiveness. PMID- 9404564 TI - The severity of clinical symptoms in ragweed-allergic patients is related to the extent of ragweed-induced complement activation in their sera. AB - We have previously reported a correlation between the extent of ragweed allergen (RWA)-induced in vitro serum complement activation and the symptom scores registered daily during the ragweed (RW)-blooming season in RW-allergic patients. The present study was performed in 22 15-17-year-old RW-allergic adolescents. Serum samples were incubated with 100 micrograms/ml RWA, and the generation of different complement activation products was measured by ELISA or RIA. Symptom scores were registered for 4 weeks during the RW-blooming season. The patients were divided according to the extent (low or high) of the generation of complement activation products, and symptom scores registered in the two groups were compared by two-way ANOVA. Significantly higher symptom scores were obtained in the high than in the low complement activation group (P values: 0.049 for C1rC1sC1inh, 0.022 for C3bBbP, 0.015 for C5b-9, 0.0001 for C3a, and 0.0008 for C5a). Similar results were obtained at the measurement performed in the sera obtained from the same patients half a year before the season (P values: 0.022 for C3bBbP, and 0.005 for C5b-9). These findings indicate that complement activation induced by the allergen may enhance the clinical symptoms of RW allergy. PMID- 9404565 TI - IgE responses to Dermatophagoides pteronyssinus native major allergens Der p 1 and Der p 2 during long-term specific immunotherapy. AB - We investigated by ELISA the IgE response to whole extract of the house-dust mite Dermatophagoides pteronyssinus (Dp) and to the native major allergens, Der p 1 and Der p 2, in sera from 18 adult patients (group A) with Dp-allergic asthma before (t0) and 1, 2, 3, and 4 (t1-t4) years after subcutaneous specific immunotherapy (SIT). A qualitative reduction (P = 0.05) of the IgE responses to Dp and Der p 2 was observed from t1 to t4, but a highly statistical significant decrease appeared at t3 (P < 0.01). With regard to Der p 1 IgE values, the immunotherapy induced a significant decrease (P < 0.01) at t3, but not before. In group A, the IgE responses to Der p 1 and Der p 2 were not correlated at t0 (rs = 0.31; P = 0.21) but were correlated at t3 (rs = 0.78; P = 0.001). We also examined sera from 14 adult patients (group B, same SIT schedule as group A) who were without respiratory symptoms at the end of the third year (t3) of Dp SIT. At this time (t3), there were no significant differences in Der p 1 and Der p 2 IgE levels between group A and group B. PMID- 9404566 TI - Links between hospital diagnoses of bronchiectasis and asthma. AB - In view of the conflicting notions of the relationship between bronchiectasis and asthma, we have analysed the use of hospital services by bronchiectasis and asthma patients and evaluated the links between these diseases, employing data from the Finnish Hospital Discharge Register of over 21 million hospitalization periods recorded in 1972-92. We conclude that asthma is common in hospitalized bronchiectasis patients and appears to be consequent upon this disease. PMID- 9404567 TI - Reduction of the mite-allergen reservoir within mattresses by vacuum-cleaning. A comparison of three vacuum-cleaning systems. AB - Mattresses are considered to be the main source of house-dust-mite (HDM) allergen. This study aimed to investigate different types of vacuum cleaners for their ability to reduce this allergen reservoir. Three types of vacuum cleaners were examined: a conventional vacuum cleaner, a water-trap cleaner, and a central vacuum cleaner. Cut out pieces of mattresses were analyzed for content of mite allergen after vacuum-cleaning. Three polyester mattresses from the homes of three children were selected for the study. These three mattresses had earlier been found to be contaminated by group 1 HDM allergen (8-15 micrograms/g of dust). Each mattress was divided into three main sections, separated by a safety zone, and each main section was subdivided into four parts. The three main sections were cleaned by each vacuum-cleaning system. Pieces 2-cm-thick of the surface of the mattresses were cut out, and extracts were made to assess the amount of mite allergen still present after different intensities of vacuum cleaning. A considerable reduction in the level of mite allergen was found after using all three of the vacuum-cleaning systems. Only 22% of the original mite allergen reservoir remained after intense vacuum-cleaning. Since the number of mattresses in the investigation was low, and niche formations of group 1 HDM allergen within the mattresses were observed, the results must be interpreted with caution. Furthermore, allergen-producing living dust mites were not affected by this method. However, intense vacuum-cleaning seems to reduce considerably the level of the mattress-bound mite-allergen reservoir, a fact which may be important in mite-allergic and mite-exposed families. PMID- 9404568 TI - Exercise tests in large groups of children are not a suitable screening procedure for undiagnosed asthma. AB - Schoolchildren (n = 473), 12-13 years of age, from five schools, and without known asthma, participated in a screening test for exercise-induced asthma (EIA). The children were tested in large groups of 10-15 pupils. Peak expiratory flow (PEF) was measured before, immediately after, and 6-8 min after 6 min of running exercises in a gymnasium. A fall in PEF of at least 10% on two separate test occasions was considered an abnormal result. Children with abnormal results were given an asthma questionnaire and then tested individually in hospital with a standardized exercise test measuring FEV1, PEF, and flow/volume curve. In the screening test, 23 (4.9%) of the 473 children had an abnormal result. When tested in hospital, five (1%) children had a decrease in PEF and/or FEV1 of at least 10% (10-14%) after exercise. Furthermore, three of these five children had a history indicating mild EIA. We conclude that the use of PEF measurement as a screening method for EIA in large groups of schoolchildren cannot be recommended because it yields many false-positive results. PMID- 9404569 TI - Anaphylactic reactions to ingested carmine (E120). AB - We report five cases of anaphylactic reaction to carmine (cochineal, E120) after patients drank an alcoholic beverage. By means of positive skin prick tests (SPT) and positive RAST to carmine. IgE-mediated sensitization could be established. One nonatopic patient showed also a great amount of serum IgE antibodies to the carmine acid-albumin conjugate. Due to its widespread use in the food and cosmetic industry, carmine should be tested in the allergy work-up in case of allergic reactions after a drink or a meal. PMID- 9404571 TI - The optimum concentration for skin prick testing. PMID- 9404570 TI - Mercurochrome allergy. Immediate and delayed hypersensitivity. AB - We describe eight patients suffering from Mercurochrome allergy. Patch and prick tests were carried out with the following organic and inorganic mercury compounds: thimerosal, Mercurochrome, phenylmercuric acetate, phenylmercuric nitrate, metallic mercury, and mercuric chloride, and with sodium fluorescein. Two patients had an anaphylactic reaction a few minutes after application of Mercurochrome. The prick tests with Mercurochrome were positive and they were negative with the other tested products. All patch tests were negative. In the other six patients, the clinical picture was local eczema, and the patch tests were all positive with Mercurochrome and the inorganic mercuric derivatives. Positive patch tests with thimerosal were found only in two patients, and only one had a positive patch test with salts of phenylmercury. In four patients, the prick test with Mercurochrome, negative in immediate reading, gave a late eczematous reaction. PMID- 9404572 TI - Primary sensitization to Morus alba. PMID- 9404573 TI - Wasp sting-associated cold urticaria. PMID- 9404574 TI - Delayed hypersensitivity to tetrazepam. PMID- 9404575 TI - Asthma to latex and amoxicillin. PMID- 9404576 TI - Possible role of IgE in acute-phase response. PMID- 9404577 TI - Anaphylactic reaction to methotrexate. PMID- 9404578 TI - Occupational allergy to rye flour in carpenters. PMID- 9404579 TI - Bone density and osteoporosis: crunching more than numbers. PMID- 9404580 TI - Treatment strategies for acute myocardial infarction: looking through the window of opportunity. PMID- 9404581 TI - Revolutionary and evolutionary progress in breast cancer. PMID- 9404582 TI - Instrument performance in bone density testing at five Australian centres. AB - AIMS: To assess the in vitro precision and accuracy of bone mineral densitometry (BMD) within and between locations at five Australian centres. METHODS: Using a multicentre reliability study the accuracy and short- and long-term precision of dual-energy X-ray absorptiometry (DXA) in vitro was compared on five instruments. Measures were performed using pencil beam mode on four Hologic QDR-2000 densitometers and one Hologic QDR-1000/W (Hologic Inc, Waltham, MA). RESULTS: Short-term precision of bone mineral density measurement was less than 0.5% for spine phantoms (n = 10 for each centre, mean intrasite coefficient of variation [CV] 0.39 +/- 0.09% [SD]) and for hip phantoms (n = 10 for each centre, mean intrasite coefficient of variation [CV] 0.34 +/- 0.10% [SD]). Between-centre measurement (n = 10 for each phantom) of a single spine phantom and a single hip phantom (specified mineral contents-58.5 g and 38.6 g, respectively) revealed ranges of bone mineral content of 57.7-58.1 g (all-point CV = 0.52%) and 37.1 37.8 g (all-point CV = 0.70%), respectively. When results from pairs of machines were compared there were statistically different mean BMD results for the majority of the ten possible pairings for both spine and hip measurements. Each study centre measured in vitro stability of phantom BMD measurements over a one year period (n = 45-283, median 157 for spine; and n = 0-262, median 38, for hip); CVs ranged from 0.38 to 0.53% for the spine measurements and from 0.38 to 0.54% for the hip measurements. The mean all-point accuracy of the spine phantom measurements was 99.1% and the hip phantom measurements was 96.7%. CONCLUSIONS: Across a number of instruments DXA demonstrates in vitro all-point precision of 0.5% for the spine phantom and 0.7% for the hip phantom. The instrument demonstrates accuracy of greater than 99% at the spine and 96% at the hip. This finding has clinical, research and quality control implications. PMID- 9404583 TI - Time delay in the treatment of acute myocardial infarction (AMI): a comparison of primary percutaneous transluminal coronary angioplasty (PTCA) with thrombolysis. AB - BACKGROUND: If primary percutaneous transluminal coronary angioplasty (PTCA) cannot be performed within times comparable to thrombolysis, the possible advantages of that management may be offset by the logistic difficulties associated with its delivery. AIM: To measure and compare the time delay involved in administration of thrombolysis and primary PTCA over a one year period and examine causes for delay greater than 60 minutes. METHOD: Prospective data collection on all patients treated with primary PTCA or thrombolysis. A quality improvement process was applied. RESULTS: Eighty-five patients were treated with thrombolysis with a delay of 39 +/- 8 (SD) minutes, 12 patients being treated more than 60 minutes after presentation. Primary PTCA was used in 79 patients with a delay of 48 +/- 12 (SD) minutes, 21 patients being treated after more than 60 minutes. Time delays in the two management groups were significantly different (p = 0.03) but that in primary PTCA during routine hours was not significantly different from that in thrombolysis treated patients (p = 0.07). Causes for revascularisation delay greater than 60 minutes from presentation are discussed. CONCLUSIONS: With appropriate facilities and organisation, patients with acute myocardial infarction presenting within normal working hours can be treated with primary PTCA without compromising their care due to time delay. Many patients managed with primary revascularisation by thrombolysis or primary PTCA with a delay of more than 60 minutes have identifiable clinically appropriate delays. PMID- 9404584 TI - Sublingual nifedipine in human pregnancy. AB - BACKGROUND: Nifedipine is widely used for acute lowering of blood pressure in obstetric hypertensive emergencies. It has not been approved for this indication or widely assessed. AIMS: To examine retrospectively the efficacy of nifedipine over a 12 month period in a high risk obstetric service. METHODS: Chart review of all patients admitted to hospital in the antenatal period with moderate to severe hypertension. Description of their management, usage of nifedipine, and pregnancy outcome. RESULTS: Sublingual nifedipine resulted in significant lowering of blood pressure without hypotension. Pregnancy outcome was satisfactory in all patients. CONCLUSIONS: Sublingual nifedipine was effective, easy to administer, and without serious complications in this retrospective study. PMID- 9404585 TI - 'Normal' lung function in rural Australian aborigines. AB - BACKGROUND: Although lung diseases are a leading cause of premature mortality in Australian Aborigines, little is known about normal lung function in these people. AIM: To develop models for 'normal' spirometric function in rural Australian Aborigines. METHOD: A cross-sectional population-based study of four rural Aboriginal communities was performed in Queensland, Northern Territory and South Australia, Australia. We studied 261 children aged seven-19 years and 332 adults aged 20-80 years who were free of symptoms and had no clinical signs of chronic lung disease. The outcome measures were forced expiratory volume in one second (FEV1) and forced vital capacity (FVC). Multiple linear regression was used to develop models for FEV1 and FVC and comparisons were made with Caucasians and indigenous people from other countries. RESULTS: The Aboriginal people studied had FEV1 and FVC values that were lower (20% and 30% respectively) than those found in Caucasians of the same height, age and gender. As a consequence, they had relatively high FEV1/FVC ratios. Those studied also had forced expiratory volumes that were lower than those found in African Americans and other indigenous peoples. CONCLUSIONS: Apparently healthy rural Aboriginal people have low forced expiratory volumes when contrasted with Caucasians and indigenous peoples such as African Americans. More research is required to determine if this is 'normal' or a product of the suboptimal environment into which many Aboriginal people are born. PMID- 9404586 TI - The value of the lipid-laden macrophage index in the assessment of aspiration pneumonia. AB - BACKGROUND: A semi-quantitative index of lipid-laden macrophages on broncho alveolar lavage (BAL) has been reported to be highly sensitive but only moderately specific for aspiration in adults. There has been little published literature evaluating this technique since the original report. AIMS: To assess the value of a lipid-laden macrophage index (LLMI) of greater than 100 to confirm the clinical diagnosis of aspiration in patients with abnormal radiological investigations. METHODS: Prospective evaluation of 80 adult patients with abnormal radiology was undertaken using BAL and the LLMI. A diagnosis of aspiration was made prior to bronchoscopy if the patient had one or more of: clinically witnessed aspiration, positive barium swallow or speech pathology assessment; or an upper gastrointestinal endoscopy showed severe reflux oesophagitis and the patient had a history consistent with aspiration. RESULTS: Eighteen patients were diagnosed with aspiration. Of these, 17 had an index > 100, and one had an index of 94 (mean 157, 99% CI 127-187, range 94-238). Of the other 62 subjects, seven had an index > 100 (mean 46, 99% CI 22-70, range 0-303). There was a significant difference between index scores for the two groups (p = 0.002). For aspiration, an index > 100 had a sensitivity of 94% and a negative predictive value of 98%. The specificity was 89%, with a positive predictive value of 71%. CONCLUSIONS: The LLMI is a sensitive indicator for aspiration causing radiological lung disease in adults. Its lack of specificity means it cannot be the sole means of diagnosis, but it can allow better targeting of other investigations in those patients in whom bronchoscopy is undertaken to investigate radiological abnormalities. PMID- 9404587 TI - Early prediction of risk in patients with suspected unstable angina using serum troponin T. AB - BACKGROUND: One-third of patients with rest angina are reported to have detectable cardiac troponin T in the serum and may be at increased risk of serious cardiac events. AIM: To investigate whether a single early estimation of serum troponin T was an independent predictor of serious cardiovascular complications in patients with suspected unstable angina. METHODS: A prospective cohort study in which patients with suspected rest angina had a serum troponin T estimation 14 hours after symptom onset and were classified using discriminator levels of serum troponin T of 0.05 and 0.1 microgram/L as well as a number of other variables. All patients were followed for six months to document any cardiac complications and a stepwise logistic regression analysis was conducted to determine independent risk factors of complications. RESULTS: One hundred and sixty-four patients were evaluated. Using a discriminator level of 0.05 microgram/L 54 patients (33%) had detectable troponin T. The admission ECG was the only independent predictor of cardiac events in hospital--odds ratio 4.0 (95% CI 1.7-9.6). Detectable troponin T did not appear to be an independent predictor of serious complications. During the six-month follow-up period, detectable troponin T using a discriminator of 0.05 microgram/L was an independent predictor of serious complications--odds ratio 3.7 (95% CI 1.8-7.6). CONCLUSIONS: In patients with suspected rest angina, detectable serum troponin T > 0.05 microgram/L is an independent predictor of serious cardiac events during the six month follow-up period although not during hospitalisation. Using a single, early serum troponin T estimation and other variables available at the time of admission, a high risk subgroup who may benefit from early investigation and revascularisation can be identified. PMID- 9404588 TI - Early management and outcome of acute stroke in Auckland. AB - BACKGROUND: Studies of acute stroke management in stroke units and tertiary referral hospitals may not accurately reflect practice within the population. Reliable information on the management of stroke within a population is sparse. AIMS: To compare clinical practice in acute stroke management in Auckland with guidelines for the management and treatment of stroke in other countries; to provide a baseline measure against which future changes in management can be evaluated. METHODS: All new stroke events in Auckland residents in 12 months were traced through multiple case finding sources. For each patient, a record of investigations and treatment during the first week of hospital admission was kept. RESULTS: One thousand eight hundred and three stroke events (including subarachnoid haemorrhages) occurred in 1761 patients in one year. Twenty-seven per cent of all events were managed outside hospital and 73% of the stroke events were treated in an acute hospital. Of the 1242 stroke events admitted to an acute hospital in the first week, only 6% were managed on the neurology and neurosurgery ward, 83% were managed by a general physician or geriatrician and 42% had computed tomography (CT). Of 376 validated ischaemic strokes, 44% were treated with aspirin and 12% with intravenous heparin. Of the 690 unspecified strokes (no CT or autopsy), 38% received aspirin and 0.5% heparin. The 28 day in hospital case fatality for all stroke events admitted to an acute hospital during the first week was 25%. CONCLUSIONS: In Auckland, management of acute stroke differed from clinical guidelines in the high proportion of patients managed in the community, the low rate of neurological consultation, and the low frequency of CT scanning. Despite these deficiencies in management, the 28 day hospital case fatality in Auckland was similar to other comparable studies which had a high proportion of cases evaluated by a neurologist and CT. PMID- 9404589 TI - The Family Atherosclerosis Risk Intervention Study (FARIS): risk factor profiles of patients and their relatives following an acute cardiac event. AB - BACKGROUND: Relatives of patients with coronary heart disease have a heightened risk of cardiovascular disease. Attendance at a family-based screening clinic after an acute cardiac event could motivate patients and relatives to modify their lifestyles. AIMS: The Family Atherosclerosis Risk Intervention Study (FARIS) aimed to determine (i) whether a high proportion of patients and relatives would attend a special screening and prevention programme; (ii) whether the risk factor profiles of relatives would be worse than those in the general community; and (iii) whether ongoing management of patients and families together in a special clinic would improve risk factor profiles. METHODS: Consecutive patients, together with spouse, siblings and offspring, aged 18 to 69 years, were randomly allocated three months after an acute cardiac event to attend a special outpatient clinic, a screening and advice group, or a control group. Risk factor measures were total cholesterol, HDL cholesterol (HDLC), systolic blood pressure (SBP), body mass index (BMI) and smoking behaviour. This paper presents the risk factor profiles of all FARIS attenders and compares those of family members, age adjusted, with risk factors measured in a multicentre urban cross-sectional survey conducted in the same period. Differences between groups were compared using t-tests for numerical variables and ANOVA and chi-square for categorical variables. RESULTS: Six hundred and twenty-eight patients and 1723 family members were enrolled, representing 85.9% and 82.7% of eligible patients and relatives respectively. Risk factors were significantly worse amongst family members than among those in the population survey. PMID- 9404590 TI - The decline in asthma hospitalisations in persons aged 0-34 years in New Zealand. AB - AIMS: Hospitalisation rates for asthma for the 0-14 year and five-34 year age ranges have been examined from 1969 to 1993 to determine whether the rise observed between the 1960s and 1980s has continued into the 1990s. RESULTS: In the 0-14 age range, hospitalisations peaked in 1986 then fell by 18.7% by 1993. There was a corresponding rise in hospitalisation rates for acute bronchitis/bronchiolitis and it is possible that the fall in asthma hospitalisations in this age range is at least partly explained by diagnostic transfer. On the other hand, the trends in the five-34 age range appear unlikely to be explained by diagnostic transfer. The rate peaked in 1986 and fell by 34.7% by 1993, with most of the decline occurring after 1989. This in part parallels the trends in mortality in this age range, which saw a sudden fall in the death rate in 1989. CONCLUSIONS: New Zealand is not only benefiting from a marked fall in asthma deaths, but is also benefiting from a marked decline in asthma hospitalisations in young adults, and probably also in children. PMID- 9404591 TI - The place of amiodarone: an overview of the four recent large controlled trials. PMID- 9404592 TI - Serious infections due to Staphylococcus lugdunensis. PMID- 9404593 TI - Whipple's disease: unusual presentations. PMID- 9404594 TI - Anomalous left anterior descending coronary artery presenting with acute myocardial infarction. PMID- 9404595 TI - An unusual cause of cyanosis (isosorbide dinitrate induced methaemoglobinaemia) PMID- 9404596 TI - CSF dissemination of a benign choroid plexus papilloma (CPP) PMID- 9404597 TI - Thrombocytopenia and hepatitis complicating ticlopidine therapy. PMID- 9404598 TI - Evidence of no difference or just no evidence of any difference. PMID- 9404600 TI - Grounds for more backup. PMID- 9404599 TI - Gastroenteritis in Australia. PMID- 9404601 TI - A case of serotonin syndrome induced by moclobemide during an extreme heatwave. PMID- 9404602 TI - Diffuse alveolar damage following a single administration of a cyclophosphamide containing chemotherapy regimen. PMID- 9404603 TI - The use of biologics in the treatment of rheumatoid arthritis (RA) PMID- 9404604 TI - Interferon therapy for acute hepatitis C viral infection--a review by meta analysis. AB - BACKGROUND: Hepatitis C viral (HCV) infection poses a major health problem for Australia. Currently interferon therapy is approved only for people with chronic infection, yet the literature contains a number of studies that show that there is a better response to interferon in symptomatic acute HCV. AIM: To review the response to interferon therapy in acute HCV by way of meta-analysis. METHODS: This study was a retrospective review of the data on the use of interferon therapy in acute HCV. The meta-analysis was performed using the methods of DerSimonian and Laird. Data were presented by calculating the risk difference which estimated efficacy by calculating the proportion of patients in treatment groups who responded better (0 to +1.0) or worse (0 to -1.0) than untreated control groups. RESULTS: A meta-analysis of six studies on the use of 3MU of interferon alpha 2b (IFN-alpha 2b) three times a week for six to 24 weeks showed a significant response as measured by long term (> 12 months) normalisation of alanine aminotransferase (ALT) and clearance of HCV RNA (as measured by polymerase chain reaction). The risk of difference was +0.31 (95% CI of +0.19 to +0.43, p < 0.01) and +0.33 (95% CI of +0.08 to +0.58, p < 0.001) respectively. Slightly better results were seen with daily doses of 3MU of interferon beta (IFN beta) given intravenously over four to seven weeks. This produced a risk difference of +0.57 (95% CI of +0.26 to +0.88, p < 0.02) for normalisation of ALT and +0.83 (95% CI of +0.61 to 1.00, p < 0.001) for clearance of HCV. Results for higher daily doses of both IFN alpha and beta were limited to a few studies and most were uncontrolled. 6MU of IFN-alpha 2b three times a week for 16 to 24 weeks produced a risk difference of +0.53 (95% CI +0.17 to +0.89, p < 0.05) for normalisation of ALT and +0.44 (95% CI +0.06 to +0.82) for clearance of HCV RNA. Results with 6MU daily for eight weeks of IFN-beta in an uncontrolled study, showed up to 90% patients cleared HCV long term. Preliminary results with 10MU of IFN-alpha 2b daily for four to six weeks also showed long term clearance of HCV RNA and normalisation of ALT in 90% of treated patients. CONCLUSION: Short term (six weeks to six months) treatment of symptomatic acute HCV with interferon (both alpha and beta) produced a better long term response rate than prolonged therapy (> 12 months) in chronic HCV. Daily doses of 6MU and 10MU produced better responses than 3MU but more studies are needed to determine the optimum regime. PMID- 9404605 TI - Microchipping: some light at the end of the tunnel. PMID- 9404606 TI - Laryngeal paralysis in a rottweiler with neuroaxonal dystrophy. PMID- 9404607 TI - Idiopathic eosinophilic meningoencephalitis in rottweiler dogs: three cases (1992 1997). AB - We report three cases of eosinophilic meningitis in young male Rottweiler type dogs in New Zealand. No cause for the disease was identified. There were variable clinical signs referable to central nervous system dysfunction, and a variable response to treatment. PMID- 9404608 TI - Pseudomonas mastitis in a dairy herd. PMID- 9404609 TI - Surgical management of a ruptured bladder secondary to a urethral obstruction in an alpaca. PMID- 9404610 TI - Use of a multifenestrated indwelling lavage system for treatment of septic digital tenosynovitis in cattle. AB - Septic tenosynovitis was diagnosed in seven cattle on the basis of history, physical examination, radiographs, cytological examination of tendon sheath fluids, and microbial culture. A commercially available indwelling multifenestrated silicone rubber drain was used to perform frequent lavage of the flexor tendon sheaths. The sepsis resolved in all cattle. Five of six cattle for which long-term (> 1 year) follow-up information was available were clinically sound on the affected limb and had remained productive members of the herd. PMID- 9404611 TI - Ovine Johne's disease. PMID- 9404612 TI - Meeting food safety challenges. PMID- 9404613 TI - Control of cattle ticks (Boophilus microplus) on Queensland dairy farms. PMID- 9404614 TI - Microsporidia (Encephalitozoon cuniculi) in wild rabbits in Australia. AB - OBJECTIVE: To determine the prevalence of infection with Encephalitozoon cuniculi in wild rabbit populations in Western Australia, and to isolate the organism from seropositive rabbits. DESIGN: Serological screening of wild and clinically affected domestic rabbit populations. SAMPLE POPULATION: Eighty-one wild rabbits from south-western Western Australia and 29 laboratory rabbits. PROCEDURE: Indirect immunofluorescence antibody technique and in-vitro amplification of parasite isolates in fibroblast cultures. RESULTS: Of the 81 wild rabbits and 29 laboratory rabbits, 20 and 22 respectively, had antibodies to E cuniculi. E cuniculi from the urine of one seropositive laboratory rabbit and from brain and kidney tissues of eight and five seropositive laboratory and wild rabbits respectively were isolated in fibroblast cultures. CONCLUSION: E cuniculi infection has been shown for the first time to be prevalent in wild rabbits in Australia. Techniques have been developed for the isolation and culture of the causative agent. Comparative studies can now be undertaken to determine risk factors for clinical disease in domestic rabbits and the relationship among E cuniculi isolates from wild and domestic rabbits. PMID- 9404615 TI - Fasciola hepatica infection in farmed emus (Dromaius novaehollandiae). AB - OBJECTIVE: To describe two cases of infection with Fasciola hepatica in young farmed emus, subacute and chronic fasciolosis and a response to treatment of the flock with albendazole. PROCEDURE: Gross lesions were found at necropsy and hepatic lesions in microscopic examination. The parasite recovered from one emu was identified by its morphological characteristics and an egg count reduction test was carried out after treatment of the flock with albendazole. RESULTS: Hepatic lesions resembling subacute and chronic fasciolosis of ruminants were identified. An adult fluke was recovered from the liver of one of the birds and was identified as F hepatica. The eggs of the fluke were irregular in shape and size. No fluke eggs were identifiable in faeces of live emus 10 days after treatment of the flock with albendazole at a dose of 10 mg/kg. CONCLUSIONS: This is the first reported case of infection with F hepatica in farmed emus and the first report of the occurrence of Fasciola infection is the class Aves. The irregular shape and size of the eggs may be attributable to infection of an aberrant host. Treatment with albendazole eliminated eggs from the faeces of the flock. PMID- 9404616 TI - Reference ranges for biochemical and haematological values in farmed saltwater crocodile (Crocodylus porosus) yearlings. AB - OBJECTIVE: To determine reference ranges for healthy yearling farmed saltwater crocodiles by performing routine biochemical and haematological laboratory tests on blood samples. DESIGN: A clinico-pathological study. PROCEDURE: Blood samples were collected from 120 healthy yearlings from four Northern Territory crocodile farms and body weight and length were measured. All animals had been fasted for 2 days before sample collection. Routine biochemical analytes were determined on 120 samples and haematological values determined on 30 samples (from one farm). RESULTS: Reference ranges for biochemical and haematological values were determined for farmed yearling saltwater crocodiles in the Northern Territory. CONCLUSION: The results were comparable with published reference ranges for other crocodilian species. Other published results of haematological values from saltwater crocodiles were from very young (6-week-old) hatchlings and older (2- to 4-year-old) crocodiles. Differences in values were presumed to be caused by age-related factors. PMID- 9404617 TI - Amoxycillin as an alternative to dihydrostreptomycin sulphate for treating cattle infected with Leptospira borgpetersenii serovar hardjo. AB - OBJECTIVE: To assess the effect of amoxycillin treatment on urinary excretion of leptospires from cattle infected with Leptospira borgpetersenii serovar hardjo. DESIGN: A chemotherapy trial with controls. PROCEDURE: Fourteen heifers serologically negative to L hardjo were inoculated with L hardjo via the conjunctival route and assessed for evidence of infection by serological, fluorescent antibody and microbiological tests. Two injections (48 h apart) of amoxycillin at a dose of 15 mg/kg were administered intramuscularly to seven heifers 6.5 weeks after infection; the remaining heifers acted as untreated controls. Later, these seven control group heifers were treated with a single dose of amoxycillin (15 mg/kg). Samples of urine were collected before and after amoxycillin treatments; kidneys were collected at slaughter, and examined by fluorescent antibody test and microbiological culture. RESULTS: Leptospires were isolated from the urine of 11 of 14 heifers inoculated with L hardjo. After treatment of six of these with two injections of amoxycillin, leptospires were not isolated. Of the controls, four of the five initially leptospiruric heifers continued to shed leptospires; after a single injection of amoxycillin, no leptospires were detected in the kidneys of these four. CONCLUSION: Amoxycillin may be an acceptable alternative to dihydrostreptomycin sulphate for the treatment of cattle infected with L hardjo. PMID- 9404618 TI - Experimental Leptospira borgpetersenii serovar hardjo infection of pregnant cattle. AB - OBJECTIVE: To observe the effect upon the foetus of experimental infection of pregnant cattle with Leptospira borgpetersenii serovar hardjo. DESIGN: A disease transmission study using pregnant cattle. PROCEDURE: Fourteen heifers serologically negative to L hardjo were artificially inseminated and later challenged with a north-Queensland isolate of L hardjo by conjunctival inoculation. The heifers were serologically monitored and their urine examined for the presence of leptospires using culture and fluorescent-antibody tests at appropriate intervals. Elective caesarean sections were performed on pregnant heifers at 6.5 weeks after the challenge. Foetuses were examined using serological, histopathological, microbiological and fluorescent-antibody tests. RESULTS: Ten of the heifers became pregnant, but three subsequently aborted before challenge. After challenge, all 14 heifers seroconverted and L hardjo was isolated from the urine of 6 of the 7 pregnant heifers. No evidence of foetal L hardjo infection was detected. Two of the foetuses had histopatho-logical lesions consistent with Neospora sp infection. CONCLUSION: It is likely that the isolate of L hardjo used in this study does not normally infect the foetus. Neospora sp may be a more significant cause of bovine reproductive wastage. PMID- 9404619 TI - Evaluation of a PCR assay for identification and differentiation of Campylobacter fetus subspecies. AB - OBJECTIVE: To evaluate a polymerase chain reaction assay for identification of Campylobacter fetus and differentiation of the defined subspecies. DESIGN: Characterisation of bacterial strains by traditional phenotyping, polymerase chain reaction, a probabilistic identification scheme and macrorestriction profiling using pulsed field gel electrophoresis. PROCEDURE: The results of identification of 99 bacterial strains as determined by conventional phenotyping or by polymerase chain reaction were compared. Two of these were type strains of C fetus subsp fetus and C fetus subsp venerealis; the remaining strains were field isolates putatively identified as C fetus. In cases where the subspecies identity was disputed, isolates were identified by means of a probabilistic identification scheme and by macrorestriction profiling. RESULTS: The agreement between strain identities initially suggested by traditional phenotypic methods and the PCR assay was found to be 80.8%. The polymerase chain reaction proved to be a reliable technique for the species and subspecies identification of C fetus; equivocal results were obtained in only two instances. Initial misidentifications by conventional phenotyping methods were attributed to methodological differences used in various laboratories. CONCLUSION: Our results indicate that misidentification of C fetus in routine diagnostic laboratories may be relatively common. The PCR assay evaluated gave rapid and reproducible results and is thus a valuable adjunctive method for the identification of C fetus and subsequent subspecies differentiation. PMID- 9404620 TI - Mycobacteriosis in young freshwater crocodiles (Crocodylus johnstoni). PMID- 9404621 TI - Dieldrin in sheep. PMID- 9404622 TI - Increase of the aversive value of taste stimuli following ibotenic acid lesion of the central amygdaloid nucleus in the rat. AB - Male Sprague-Dawley rats received bilateral ibotenic acid lesions of the central amygdaloid nucleus (CeA) and were compared to sham-lesioned rats in their response to different concentrations of saccharin and quinine solutions. In two bottle choice test situation, the lesioned rats exhibited a lower saccharin preference at concentrations of 2.5; 7.5 and 25 mM, while their aversion towards quinine and the highest concentration of saccharin (50 mM) was increased. In a one-bottle test, the lesioned rats showed consistent decreases in their consumption of 2.5 and 7.5 mM saccharin solutions whereas their intake of 0.9 mM solution of saccharin was equal to that of the sham-lesioned rats. The lesion of the CeA had no significant effects on the acquisition of conditioned taste aversion. There was less postoperative weight gain in lesioned rats as compared to sham-lesioned animals but the lesion had no significant effect on daily water intake. These findings suggest that the CeA plays an important role in the normal response to exteroceptive food stimuli via modulation of the aversive value of taste stimuli. The results are discussed in the context of an interaction between the CeA and the lateral hypothalamus (LH) in the modulation of palatability and feeding behavior. PMID- 9404623 TI - The ground reaction forces of postural adjustments during skilled reaching in unilateral dopamine-depleted hemiparkinson rats. AB - Rats with unilateral dopamine (DA) depletions (hemiParkinson analogue rats) produced by intracerebral 6-hydroxydopamine injection are impaired in using the contralateral (bad) limbs for postural adjustments. This article examines whether the bad limbs are impaired in applying the forces required to initiate postural adjustments that anticipate and accompany voluntary movements. The rats were trained to reach for food using their good paw while standing on small platforms, each of which measured force changes produced by an individual limb. In one condition the force platforms were aligned to support the limb placement of normal rats and in the second they were aligned to permit the DA-depleted rats to use a compensatory reaching stance. It was found that the bad limbs of the DA depleted rats produced normal supporting reactions but did not initiate adjustments in posture. Postural adjustments were initiated with the good limbs and preceded rather than accompanied the reaching movements. When constrained to use the posture of normal rats, the DA-deplete rats could not reach successfully, but when allowed to adjust their stance to increase reliance on the good limbs, reaching performance improved. Measures of ground reaction forces confirm that DA depleted rats can support posture but cannot initiate postural adjustments with their impaired limbs. PMID- 9404624 TI - Importance of the medial amygdala in rat penile erection evoked by remote stimuli from estrous females. AB - Effects of medial amygdala lesions (MAL) were examined on rat penile erection in three different experimental situations. Only sexually vigorous males, as identified by preoperative mating tests, were used. Bilateral radiofrequency lesions were confined to the posterior medial amygdala, with little systematic damage to anterior medial amygdala or to adjacent structures. Lesion electrodes were withdrawn without current application in sham-operated animals (SHAM). After recovery for brain surgery, males were tested for (1) noncontact erection (NCE) that occurs when males were placed in proximity to inaccessible estrous females, (2) reflexive erection evoked in supine males by retraction of the penile sheath, and (3) copulatory behaviour with receptive females. In the NCE test, none of the MAL males showed penile erection during the 20 min observation, whereas 70% of the SHAM males showed it (P < 0.001). In contrast, no erectile dysfunction in the MAL males was detected in the other two tests. MAL males displayed more penile body erections (flips) than SHAM males in the reflexive-erection test (P < 0.05). In the copulation test, most of the MAL males achieved intromission, but their intromission ratio, a partial measure of erectile function, was marginally lower than that of SHAM males (P = 0.051). MAL males had longer intervals between intromissions (P < 0.001); as a result, none of them ejaculated during the 20 min period that followed the first intromission. The results suggest that the posterior medial amygdala plays an essential role in the regulation of NCE, and it may also contribute to the regulation of erection in other contexts. PMID- 9404625 TI - Visually induced motion perception and visual control of postural sway in congenital nystagmus. AB - In congenital nystagmus (CN) the threshold for detecting motion of visual objects is increased. To determine whether this increase is due to a deterioration of visual motion signals or whether visual-vestibular interactions (which are necessary to judge object-motion in space) are also involved we examined how CN patients use visual motion signals to evaluate self-motion in perceptual and behavioral tasks. Using an optokinetic drum we measured the minimum optokinetic acceleration necessary to induced motion perception of the visual environment in CN patients. This threshold was significantly elevated in the CN patients compared with normals (20.1 deg/s2 to 3.25 deg/s2). We further addressed the question whether the elevation of this threshold is due to a deficiency in evaluating visual motion in general or to a specific modification affecting the percept of visual object-motion with respect to the inertial reference only. We thus measured the latency of visually induced self-motion perception, which was found to be very similar or even slightly smaller (1.7 +/- 0.7 s) compared with normals (2.2 +/- 1.7 s). Moreover, subjects with CN were found to use vision quite efficiently for the visual stabilization of posture (Romberg quotient 2.0 +/- 1.16), even if they did not reach the level of normals (Romberg quotient 3.7 +/- 1.1). The results indicate that CN affects the estimate of object-motion in a specific and much more severe way than the estimate of self-motion. The minimal effect of CN on self-motion perception can be explained by the low pass characteristics of the optokinetic input to self-motion perception. The specific deficiency in detecting object-motion indicates that adaptation to CN occurs on the level of visual--vestibular interactions for the perception of visual object motion and not on the level of visual motion signals. PMID- 9404626 TI - Hippocampal formation is involved in movement selection: evidence from medial septal cholinergic modulation and concurrent slow-wave (theta rhythm) recording. AB - Hippocampal rhythmical slow-wave field activity which occurs in response to sensory stimulation is predominantly cholinergic (atropine-sensitive theta rhythm), can precede movement initiation, and co-occurs during non-cholinergic theta rhythm associated with ongoing movement (atropine-resistant). This relationship suggests that theta rhythm plays some role in movement control. The present naturalistic experiments tested the idea that atropine-sensitive theta rhythm plays a role in sensory integration and planning required for initiating appropriate movements. One of a pair of hungry rats, the victim, implanted with hippocampal field recording electrodes, a septal injection cannula, and a posterior hypothalamic stimulating electrode, was given food which the other, the robber, tries to steal. Since the victim dodges from the robber with a latency, distance, and velocity dependent upon the size of the food, elapsed eating time, and proximity of the robber, the movement requires sensory integration and planning. Although eating behavior seemed normal, atropine-sensitive theta rhythm and dodging were disrupted by an infusion of a cholinergic antagonist into the medial septum. When the victim in turn attempted to steal the food back, Type 1 theta rhythm was present and robbery attempts seemed normal. Prior to cholinergic blockade, posterior hypothalamic stimulation produced theta rhythm and dodges, even in the absence of the robber, but following injections, atropine-sensitive theta rhythm and dodging were absent as the animals dropped the food and ran. The results provide the first evidence to link atropine-sensitive theta rhythm and hippocampal structures to a role in sensory integration and planning for the initiation of movement. PMID- 9404627 TI - The effects of neurotoxic lesions of the perirhinal cortex combined to fornix transection on object recognition memory in the rat. AB - The effects of lesions centred in the perirhinal cortex region (Prh) or in both the perirhinal cortex region and the fornix (Prh + Fx) were assessed in two different working memory tasks, one spatial the other nonspatial. For the spatial task the rats were tested in an eight arm radial maze, using a standard procedure in which they were rewarded for avoiding previously visited arms. The Prh + Fx, but not the Prh, rats produced significantly more errors (re-entries) and these started significantly earlier in each session when compared with a surgical control group. The nonspatial task was a test of spontaneous object recognition in which rats were tested on their ability to discriminate between a familiar and a novel object. For the initial tests the Prh group failed to discriminate between the objects, but the Prh + Fx group showed a clear preference for the novel object. Observation of the test showed, however, that the Prh + Fx group were spending a greater length of time initially exploring the sample (familiar) object. When the amount of exposure to the sample object was limited to either 20 or 40 s (i.e. was the same for all three groups), the Prh + Fx group now failed to discriminate between the two objects. This change was especially evident for shorter sample duration (20 s). The Prh group did, however, show an amelioration of their deficit with this further testing. The present results support previous dissociation between spatial and nonspatial working memory, and indicate that there may be some recovery of function following perirhinal cortical damage. PMID- 9404628 TI - Rapid decay of cocaine-induced behavioral sensitization of locomotor behavior. AB - Sprague Dawley rats received three daily intraperitoneal (i.p.) injections of saline or 15 mg/kg cocaine. Following an interval of 2, 5 or 8 days, the behavioral response of separate groups of rats to a challenge injection of cocaine (15 mg/kg) was tested in an open field. After repeated cocaine (15 mg/kg) injection, movement in both the vertical and horizontal plane was increased in cocaine-treated rats 2, but not 5 or 8, days after treatment as compared to saline-treated subjects. In addition, behavioral ratings along an ordinal scale designed to reflect increases in behavioral activation were increased in cocaine treated rats 2, but not 5 or 8, days after treatment. These results stand in contrast to other reports demonstrating long-lasting neural and behavioral changes after similar treatment regimens. Taken together, the results suggest that a treatment regimen of 15 mg/kg per day of cocaine for 3 days produces behavioral sensitization of locomotor behavior; however, this cocaine-induced behavioral sensitization does not persist beyond a few (< 5) days after repeated cocaine treatment, using the current experimental parameters. PMID- 9404629 TI - The effect of nonspatial water maze pretraining in rats subjected to serotonin depletion and muscarinic receptor antagonism: a detailed behavioural assessment of spatial performance. AB - A detailed behavioural analysis of water maze spatial performance in the rat was utilized to determine the effect of single and combined administration of p chlorophenylalanine (PCPA; 1000 mg/kg, i.p.), an inhibitor of serotonin biosynthesis, and scopolamine hydrobromide (SCO; 1.0 mg/kg, i.p.), a muscarinic receptor antagonist. In some groups a water maze pretraining regimen known as non spatial pretraining (NSP) was used to familiarize the animals with the general requirements of the task before spatial training was begun. The results showed that: (a) depletion of serotonin with PCPA had no effect on water maze performance and produced no sensorimotor disturbances; (b) antagonism of muscarinic receptors produced impairments in spatial and sensorimotor function in naive rats but neither effect was observed in rats first given NSP; (c) combined disruption of muscarinic and serotonergic function produced a severe deficit in spatial performance that was only partially alleviated by NSP; and (d) there was an association between poor maze acquisition scores and a high incidence of sensorimotor dysfunction. In addition to the water maze task the rats were also assessed for motoric performance on a beam walking test. The role of cholinergic and serotonergic systems in learning and memory is discussed. PMID- 9404630 TI - Anxiolytic actions of diazepam, but not of buspirone, are influenced by gender and the endocrine stage. AB - The effect of diazepam (1.0 mg/kg, i.p.) and buspirone (5.0 mg/kg, i.p.) on the burying behaviour latency (denoting actions on the animals' reactivity) and on the cumulative burying behaviour (directly reflecting the experimental anxiety levels), were analyzed in male-, intact females, at proestrus and metoestrus, and in neonatally-androgenized-rats. Androgenization was performed by injecting 60 micrograms/rat of testosterone propionate on day 5 after delivery. Two main groups of neonatally-androgenized rats were established: A group of animals showing permanent oestrus from the vaginal opening (acyclic females) and a group presenting the delayed anovulatory syndrome. Diazepam produced a clear reduction in experimental anxiety in males and neonatally-androgenized-females. Particularly important was the anxiolytic effect of diazepam on acyclic females that was accompanied by a significant increase in burying behaviour latency. Conversely, buspirone induced a clear reduction in burying behaviour, without modifying its latency, in all groups regardless of the gender and the neonatal treatment. Data are discussed on the basis of the androgen participation on the anxiolytic drug effects. A possible age-related benzodiazepine actions in females is suggested. PMID- 9404631 TI - Hippocampal lesions alter conditioning to conditional and contextual stimuli. AB - The control of conditioned fear behaviour by a conditional stimulus (CS) and contextual stimuli (CXT) was compared in rats with lesions to the hippocampus (HPC) or neocortex (CO), and operated controls (OC). After classical fear conditioning in a distinctive context, rats were subsequently tested in the presence of the CS and CXT (CS + CXT), the CS alone (CS-only), or context alone (CXT-only). Two experiments were conducted in which conditioned fear was measured by an active avoidance response (experiment 1) or by response suppression (experiment 2). Groups did not differ in acquiring the conditioned fear response, as measured in the CS + CON test but, in both experiments, hippocampal (HPC) groups exhibited more conditioned fear behaviour than controls in the CXT-Only and CS-Only conditions. It was suggested that control rats conditioned the fear response to a stimulus complex that incorporated the CS and CTX. Rats with HPC lesions did not form this association between the stimulus elements; instead they segregated the CS and CXT and formed independent associations between the conditioned response (CR) and each component. In showing that HPC damage disrupts the process of forming associations between environmental stimuli and that the effect is not restricted to contextual cues, the results help to resolve apparently contradictory findings regarding the role of HPC in contextual information processing. PMID- 9404632 TI - Behavioural and intestinal responses to novelty in rats selected for diverging reactivity in the open field test. AB - There are indications that the severity of functional gastrointestinal disturbances in humans is linked to individual coping styles. In rodents, the open field test can be used to assess individual differences in behavioural responsivity to novel challenges. Two groups of Wistar rats were selected for high (HA) and low (LA) locomotor activity in a novel open field and fitted with electrodes on the proximal colon. During subsequent exposure to a novel box, a smaller locomotor activation in LA was accompanied by a greater increase in colonic spike burst activity compared to HA rats, even though this novel stressful challenge did not result in a clear defecation response in either group. In contrast, no marked behavioural differences between HA and LA were seen in the shock prod paradigm. Although detection of divergent behavioural responsivity in HA and LA rats may depend on stimulus quality or intensity, combined use of behavioural selection and intestinal motility recording in freely moving rats may offer a model to study individual vulnerability to stress-related disturbances of intestinal function. PMID- 9404633 TI - Short-term memory for food reward magnitude: the role of the prefrontal cortex. AB - Memory for magnitude of reinforcement was assessed in rats using a go/no-go short term memory paradigm. During the task's study phase rats were given a piece of cereal comprised of either 25 or 50% sugar. For all trials, one of the cereal types was designated positive, the other negative. On the ensuing test phase the rat was presented with an object which covered a food well. If a positive food reward was given during the study phase, a second food reward was placed beneath the object. No food reward was placed under the object if the study phase consisted of a negative food reward. Latency to object displacement was used as the measure of performance. Following the establishment of a significant difference between latency to approach the object with reward compared to latency to approach the object without reward, rats were given either agranular insular cortex, anterior cingulate cortex, pre- and infralimbic cortex or control lesions. Agranular insular cortex lesioned animals demonstrated a mild post surgery impairment. Trials consisting of 10 and 20 s delays between the study and test phases were then introduced. Delays exacerbated the previous deficit of the agranular insular cortex lesion group, but had little effect on the other lesion groups. All animals transferred to a new set of cereals containing 25 and 50% sugar and exhibited taste preferences to sugar solutions of different concentrations. These results indicate that the agranular insular cortex may play an important role in the processing of affect-laden information within a prefrontal cortex short-term or working memory system. PMID- 9404634 TI - Behavioral alterations induced by an escalating dose-binge pattern of cocaine administration. AB - Stimulant-induced psychosis and addiction are most commonly associated with a pattern of repeated high dose 'binge' exposures, preceded by progressively escalating doses (ED). We have recently reported that an ED/multiple binge treatment with amphetamine or methamphetamine results in a unique behavioral profile, characterized by a differential change in the relative expression of locomotion and stereotypy and the emergence of a bursting pattern of ambulation. To examine the generality of this behavioral profile, a similar regimen (ED, 2-15 mg/kg; binges, 15 mg/kg x 5 hourly injections x 15 days) was used to characterize the response to cocaine. Our results show that the primary characteristics of the behavioral profile previously found with the amphetamines were also apparent with cocaine. That is, the locomotor response was significantly greater, and was characterized by a bursting pattern. These observations lend further support to the notion that the progressive behavioral changes and the underlying neurochemical alterations produced by this treatment may be implicated in the addiction and psychosis which is associated with high dose stimulant abuse. PMID- 9404635 TI - Neurotoxic lesions of the dorsal hippocampus and Pavlovian fear conditioning in rats. AB - Electrolytic lesions of the dorsal hippocampus (DH) produce deficits in both the acquisition and expression of conditional fear to contextual stimuli in rats. To assess whether damage to DH neurons is responsible for these deficits, we performed three experiments to examine the effects of neurotoxic N-methyl-D aspartate (NMDA) lesions of the DH on the acquisition and expression of fear conditioning. Fear conditioning consisted of the delivery of signaled or unsignaled footshocks in a novel conditioning chamber and freezing served as the measure of conditional fear. In Experiment 1, posttraining DH lesions produced severe retrograde deficits in context fear when made either 1 or 28, but not 100, days following training. Pretraining DH lesions made 1 week before training did not affect contextual fear conditioning. Tone fear was impaired by DH lesions at all training-to-lesion intervals. In Experiment 2, posttraining (1 day), but not pretraining (1 week), DH lesions produced substantial deficits in context fear using an unsignaled shock procedure. In Experiment 3, pretraining electrolytic DH lesions produced modest deficits in context fear using the same signaled and unsignaled shock procedures used in Experiments 1 and 2, respectively. Electrolytic, but not neurotoxic, lesions also increased pre-shock locomotor activity. Collectively, this pattern of results reveals that neurons in the DH are not required for the acquisition of context fear, but have a critical and time-limited role in the expression of context fear. The normal acquisition and expression of context fear in rats with neurotoxic DH lesions made before training may be mediated by conditioning to unimodal cues in the context, a process that may rely less on the hippocampal memory system. PMID- 9404636 TI - Fos-like immunoreactivity in the caudal diencephalon and brainstem following lateral hypothalamic self-stimulation. AB - Fos immunohistochemistry was used to stain neurons in the caudal diencephalon, midbrain and hindbrain driven by rewarding stimulation of the lateral hypothalamus (LH). Increases in Fos-like immunoreactivity were most pronounced ipsilateral to the site of stimulation and tended to be confined within discrete structures such as the posterior LH, arcuate nucleus, ventral tegmental area (VTA), central gray, dorsal raphe, pedunculopontine area (PPTg), parabrachial nucleus, and locus coeruleus. At least two of these structures, the VTA and PPTg, have been implicated in medial forebrain bundle self-stimulation. PMID- 9404637 TI - The impact of physiological insulin concentration and depletion on the metabolism of glucose, endogenous glycogen, and triglycerides in the isolated perfused heart. AB - In isolated nonworking hearts perfused with substrate-free medium for 90 min, 2000 microU insulin.mL-1 reduced the rates of glycogenolysis and lipolysis, whereas 50 microU insulin.mL-1 reduced the rate of lipolysis with no significant effect on glycogenolysis. Glycogen and triglycerides were the chief energy sources during the course of depletion in the presence and absence of insulin. In hearts perfused with medium containing 5 mM glucose for 3 h, glycogen and triglycerides contributed to the heart energy requirement, but in the presence of insulin (50 or 2000 microU.mL-1) endogenous triglycerides contributed to the heart energy requirement with net glycogen synthesis. In hearts preperfused with substrate-free medium for 90 min and perfused for another 90 min with medium containing 5 mM glucose, the depletion of endogenous glycogen and triglycerides stimulated the utilization of glucose and the synthesis of glycogen and triglycerides. In hearts preperfused with substrate-free medium for 90 min and perfused for another 90 min with medium containing glucose (5 mM) and insulin (50 or 2000 microU.mL-1), insulin enhanced the utilization of glucose and the synthesis of glycogen and triglycerides. Glucose utilization, glycogenolysis, lipolysis, and synthesis of glycogen and triglycerides were sensitive to the physiological insulin concentration and were also affected by the heart content of glycogen and triglycerides, in the isolated nonworking perfused rat heart. PMID- 9404638 TI - The polarity of tyrosine 67 in yeast iso-1-cytochrome c monitored by second derivative spectroscopy. AB - The average tyrosine polarity of 10 yeast iso-1-cytochrome c proteins and two horse heart cytochrome c proteins was assayed by second derivative spectroscopy. Yeast iso-1-cytochrome c contains five tyrosines, one of which (tyrosine 67) is in the heme pocket. The wild-type protein and the Y67F, N52V Y67F, and N521 Y67F proteins were used to differentiate events that were occurring in or near the heme pocket from those occurring closer to the protein's surface. The wild-type protein shows a substantial change in the second derivative spectrum as the protein goes from oxidized to reduced; mutants lacking tyrosine 67 do not show this change. This indicates that it is primarily the spectrum of tyrosine 67 that changes as the protein cycles between the oxidized and reduced state. One thing that contributes to the overall polarity of the heme pocket is a water molecule hydrogen bonded to several of the nearby residues. The wild-type protein has one water molecule in the heme pocket but this can be increased or decreased by introducing mutations into the protein. N52A has two water molecules and N52I has no water molecule in the pocket. The three proteins allowed us to assess the contribution of water to the inferred heme crevice polarity. The number of water molecules in the crevice correlates with the perceived polarity of the pocket when one takes account of the fact that the second water molecule in the crevice of the N52A mutant takes the position and hydrogen bonding pattern of the amide it replaces. PMID- 9404639 TI - Structural characterization of the serotype O:5 O-polysaccharide antigen of the lipopolysaccharide of Escherichia coli O:5. AB - The structure of the antigenic O-polysaccharide component of the smooth lipopolysaccharide produced by Escherichia coli serotype O:5 was investigated by composition, methylation, and periodate oxidation methods, and by 1D and 2D nuclear magnetic resonance spectroscopy. The antigenic O-chain was determined to be a high molecular weight polysaccharide composed of repeating tetrasaccharide units containing 2-acetamido-2-deoxy-D-galactose, D-galactose, D-ribose, and 3 acetamido-3,6-dideoxy-D-glucose and having the structure -[-4)-beta-D-Galp-(1-3) alpha-D-GalpNAc-(1-4)-beta-D-Quinp3NAc-(1- 3)-beta-D-Ribf-(1-]-. PMID- 9404640 TI - Pyrimidine-purine-pyrimidine triplex DNA stabilization in the presence of tetramine and pentamine analogues of spermine. AB - The formation and stability of triplex DNA were investigated in the presence of a number of tetramine (+4) and pentamine (+5) derivatives of spermine with altered spacing between the positive charges and bis(ethyl) substitution of pendant amino groups. Thermal denaturation profiles were measured for the duplex and triplex forms of poly[d(TC)].poly[d(GA)] and poly(dA).poly(dT); in both cases the pentamines were more effective than the tetramines in increasing the melting temperature (Tm) of the triplexes. Some structural effects were evident, although bisethylation of the polyamines had only a minor effect on the Tm of pyrimidine purine-pyrimidine triplexes. Relative association constants to poly(dT).poly(dA).poly(dT) and poly[d(AT)] were measured by an ethidium competition assay. These results demonstrated tighter binding of the pentamines by a factor of up to 10-fold, but bisethylation consistently decreased the relative association constants to the triplex. A third assay involving transmolecular triplex formation between separated pyrimidine-purine tracts in plasmid DNA was also employed. Again the pentamines promoted triplex formation at lower concentrations than the tetramines but structural effects were very important in determining the degree of triplex formation. These results may be important for the design of suitable ligands to stabilize triplex DNA in antigene therapeutics and to elucidate the mechanism of action of polyamine analogues as antitumor drugs. PMID- 9404641 TI - Purification and properties of glutamine synthetase from the cellular slime mould Dictyostelium discoideum. AB - Glutamine synthetase (GS) from the cellular slime mould Dictyostelium discoideum was purified to apparent electrophoretic homogeneity with a final yield of 21.7%. The native enzyme appeared to be a GS-II type enzyme. SDS-PAGE of the final preparation revealed a single band of 43.5 kDa. The enzyme has a native molecular mass of 376 kDa, determined using Superose 6, indicating that the enzyme is likely to be an octamer of identical subunits. Dictyostelium discoideum GS has an optimal temperature of 42.5 degrees C, although it is thermolabile in the absence of L-glutamate and (or) Mg(2+)-ATP. The enzyme exhibits a K(m) for L-glutamate, ATP, and NH4Cl of 2.18, 0.18, and 0.11 mM, respectively, in the L-glutamine synthetic reaction with an optimal pH of 7.9. GS from D. discoideum does not appear to be significantly inhibited by various end products of L-glutamine metabolism, although it is potently inhibited by methionine sulphoximine. These properties are those expected for an enzyme for which the primary function is the assimilation of ammonia. PMID- 9404642 TI - Pre-steady-state and steady-state function of the ileal brush border SO4(2-)-OH- exchanger. AB - Rapid-sampling analysis of the detailed time course of 35SO4(2-) uptake under pH gradient (pHin = 7.5; pHout = 5.5) conditions converged to a model of an initial burstlike pre-steady-state with relaxation to linear steady-state uptake across pig ileal brush border vesicles. A model of low affinity transport (K(m) = 7.7 mM) plus an unsaturable component described the steady-state component of 35SO4(2 ) uptake. Steady-state transport was maximal under pH-gradient conditions and sensitive to inhibition by 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid. Significant steady-state 35SO4(2-) transport sensitive to 4,4' diisothiocyanostilbene-2,2'-disulfonic acid was found under acidic (pHin = pHout = 5.5) and neutral (pHin = pHout = 7.5) pH-equilibrated conditions. Varying conditions from pH gradient to neutral pH equilibrated had no effect on the amplitude of the pre-steady-state burst or on the time constant for relaxation from pre-steady-state to steady-state conditions. However, maximal amplitude was obtained under acidic pH-equilibrated conditions. These results are incorporated into a model for the low affinity ileal brush border SO4(2-)-OH- exchanger whereby, under 0-trans (0 internal substrate concentration) conditions, translocation of inner-facing to outer-facing conformations defines the overall rate-limiting step of the transporter. Provision for slippage of the unloaded carrier could account for 35SO4(2-) transport under acidic pH-equilibrated conditions. PMID- 9404643 TI - Proteolytic profile of recombinant pro-opiomelanocortin in embryonal carcinoma P19 cells: conversion to beta-lipotropin and secretion are inhibited following incubation with canavanine. AB - A variety of proteins and peptides are produced through limited proteolysis of precursors at paired basic residues. This proteolytic bioactivation is carried out by subtilisin-like proteases, called convertases. The mRNAs of several convertases are expressed during prenatal life as well as in P19 embryonal carcinoma cells, which are a model of the totipotent cells of the embryo before and at the time of implantation. To determine whether converting activities accompany convertase mRNA expression in the early embryo, we transferred the gene of pro-opiomelanocortin (POMC) into P19 cells, by lipofection, and searched for the presence of mature peptides by high-performance liquid chromatography and radioimmunoassay techniques. In P19 cells, POMC, a precursor of several endocrine peptides, is mainly processed to beta-lipotropin rather than to beta-endorphin, both peptides having been identified by their immunoreactivity, polarity, and molecular size. These results indicate that converting capacities appear early in the embryo and that they are more similar to the activity of furin and of convertase PC1 than that of convertase PC2 in their cleavage selectivity of POMC sites. Efficiency of POMC processing can reach 50%, suggesting that convertases, with other proteases, can have an important role in ontogenesis. As for other peptide precursors in endocrine cells, the conversion of POMC in P19 cells was inhibited by the biosynthetic replacement of its arginine residues by the analog canavanine. However, the incorporation of canavanine into P19 cells also inhibited peptide secretion, suggesting that inhibition of conversion in these cells as well as in endocrine cells could indirectly result from the impairment of intracellular traffic and not only from a direct inhibition of the converting activity. PMID- 9404644 TI - Identification of 5' flanking sequences that affect human pancreatic cholesterol esterase gene expression. AB - Pancreatic cholesterol esterase (CEL) is shown to play a significant role in cholesterol metabolism. As the hydrolytic property of CEL is important for transport of lipid esters, the extent of its expression is an important factor in the metabolism of lipids. Therefore, to identify the elements that modulate the transcription of its mRNA, we obtained several cosmid clones carrying the CEL gene. From one of these cosmid clones a 6.5-kb SmaI fragment that hybridizes to the 5' untranslated region of CEL cDNA was subcloned. Primer extension and S1 protection assays revealed that the 5' untranslated region is relatively short (only 20 nucleotides long). An analysis of the 5' flanking sequence revealed typical TATA and CCAAT boxes that impart tissue specificity. Further, consensus sequences of several cis elements described earlier could also be detected in this region. To identify the promoter sequences, various deletion constructs of the 5' region were made using polymerase chain reaction. These constructs were subcloned into a bacterial plasmid vector carrying chloramphenicol acetyltransferase (CAT) as the reporter gene and transfected into HepG-2 cells. CAT activity in the cell homogenate of the transfected cells was measured 48 h after transfection. Results showed that the promoter activity of human pancreatic CEL mRNA is in a large segment of 5' flanking sequences spanning the -10 and -930 nucleotides of its gene. PMID- 9404645 TI - Partial transfection of liver with a synthetic cholesterol 7 alpha-hydroxylase transgene is sufficient to stimulate the reduction of cholesterol in the plasma of hypercholesterolemic mice. AB - The effect of administering a synthetic transgene encoding cholesterol 7 alpha hydroxylase (cyp7) on plasma cholesterol metabolism of intact mice was investigated. The synthetic cyp7 transgene (Tg1) was constructed by placing the cDNA sequence encoding the full-length cyp7 polypeptide under the control of a heavy metal inducible metallothionein promoter. The transgene was complexed with asialoorosomucoid-polylysine conjugate and introduced into mice via the tail vein. Cell marking experiments using a beta-galactosidase (lacZ) transgene as a tag showed that 5-10% of the liver can be transfected by this procedure. Administration of the Tg1 transgene to older hypercholesterolemic chow-fed mice resulted in about a 50% reduction of plasma cholesterol, regardless of whether or not transgene expression was induced by zinc treatment. In diet-induced hypercholesterolemic mice, the reduction (20%) in total plasma cholesterol was seen only when transgene expression was induced, and this reduction was due primarily to a decrease in non-high-density lipoprotein cholesterol. The maximum reduction was evident at 6 days after the introduction of the transgene and was no longer evident after 9 days. Introduction of the Tg1 transgene into young chow fed mice had no effect on the already low levels of plasma cholesterol. However, compared with the no-transgene and lacZ transgene controls, the gallbladder bile acid content of Tg1-treated mice was increased. The results show that non-viral mediated delivery of a synthetic transgene encoding cyp7 to a subpopulation of hepatocytes in the liver of intact hypercholesterolemic mice is sufficient to facilitate the temporary reduction of plasma cholesterol content. PMID- 9404646 TI - Contact of sarcoma cells with aligned fibroblasts accelerates their displacement: computer-assisted analysis of tumour cell locomotion in co-culture. AB - The shape and locomotion of rat sarcoma XC cells on glass, polystyrene, and confluent monolayer cultures of aligned human skin fibroblasts were studied with quantitative, computer-assisted methods. The cell shape depended upon the substratum; the sarcoma cells seeded on fibroblasts assumed polarized shapes. The tumour cells emigrating from aggregates and in sparse cultures showed random locomotion when plated on glass or on the polystyrene surface of tissue culture dishes in isotropic conditions. However, when sarcoma cell aggregates were plated onto underlying aligned fibroblasts, the sarcoma cells showed contact guidance, migrating along the long axes of fibroblasts. Simultaneously, suppression of migration normal to the axis of fibroblasts orientation was observed. The sarcoma cells displaced a few times faster on aligned fibroblasts than under isotropic conditions in control cultures. This fast displacement was found to result from the less frequent cell turnings and straightening of cell trajectories (i.e., from klinokinesis), and not from an acceleration of cell movement and the longer cell tracks (i.e., not from orthokinesis). The presented results support the suggestion of Abercrombie (M. Abercrombie. 1979. Nature (London). 281: 259-262.) that tumour cells may be guided by the underlying normal cells when invading surrounding tissues and forming metastases. PMID- 9404647 TI - Isolation, spreading, locomotion on various substrata, and the effect of hypotonicity on locomotion of fish keratinocytes. AB - The paper describes improved methods for the isolation of fish skin keratinocytes, which spread and locomote 15 min after trypsinization, in the absence of extracellular matrix proteins. The random locomotion of these keratinocytes under isotropic conditions on glass, plastic (polystyrene), and glass covered with poly-L-lysine or collagen IV was studied with computer-aided methods. Several methods for quantitative description of random cell locomotion were compared. The values of some parameters commonly computed showed non Gaussian distribution. A comparison of keratinocyte locomotion under isotonic and hypotonic conditions revealed that the hypotonic conditions increased cell displacement (net migration) owing to the klinokinetic and not the orthokinetic effect. PMID- 9404648 TI - Listeriolysin O potentiates immunotoxin and bleomycin cytotoxicity. AB - Antitumor immunotoxins were formed by covalently attaching the ribosome inactivating protein ricin A chain (RA) to the antitumor antibodies BR96 and L6. In vitro cytotoxicity assays established that BR96-RA was cytotoxic to H2987 human lung adenocarcinoma cells (IC50 = 6 nM), while L6-RA exhibited very low levels of cytotoxic activity (18% cell kill at 67 nM). The virulence factor from the intracellular pathogen Listeria monocytogenes, listeriolysin O (LLO), was able to potentiate the cytotoxicity of BR96-RA and L6-RA by 120- and > 1340-fold, respectively, resulting in IC50 values of approximately 50 pM. LLO also potentiated the cytotoxicity of the peptide anticancer drug bleomycin by a factor of > 2500 but had no effect on the cytotoxic activities of the anticancer drugs cytarabine and etoposide phosphate. In addition, LLO did not potentiate the cytotoxic activity of unconjugated ricin A chain or L6-RA on H2987 cells that were saturated with L6 prior to conjugate treatment. These results are attributed to LLO-induced alteration of the intracellular trafficking of molecules that are incorporated into acidic vesicles. PMID- 9404649 TI - Synthesis and enzymatic stability of phosphodiester-linked peptide oligonucleotide hybrids. AB - Nucleopeptides Ac-Tyr(p3' dACGT)-Ala-Phe-Gly-NH2, Ac-Thr(p3'dACGT)-Ala-Phe-Gly OH, Ac-Ser(p3'dACGT)-Ala-Phe-Gly-OH, and Phac-Hse(p3'dACGT)-Ala-Phe-Gly-OH, in which the 3'-end of a tetradeoxyribonucleotide is linked by a phosphodiester bond to a hydroxylated amino acid, were synthesized using a stepwise solid-phase methodology to study the influence of the linking amino acid on their stability to 3'-exonucleases. HPLC analysis of the reaction crudes after treatment of each nucleopeptide with snake venom phosphodiesterase showed that the lability of the amino acid-nucleoside linkage increases in the order Thr < Ser < Hse < Tyr. PMID- 9404650 TI - Synthesis, DNA binding, and cleaving properties of an ellipticine-salen.copper conjugate. AB - The synthesis of a DNA-cutting agent that conjugates an ellipticine chromophore and a copper complex of bis(salicylidene)ethylenediamine, referred to as a salen, is reported. The presence of the salen.Cu complex allows cleavage of DNA via oxygen-based radicals, and the ellipticine moiety serves as a DNA anchor. Spectroscopic measurements indicate that the intercalation geometry of the ellipticine chromophore is preserved with the hybrid. The cleavage is much more efficient with the conjugate than with the Schiff base copper complex alone. PMID- 9404651 TI - Synthesis and characterization of high-molecular mass polyethylene glycol conjugated oligonucleotides. AB - The use of synthetic oligonucleotides as possible drugs for human therapy is usually hampered by their low in vivo stability and capacity to achieve high concentrations at their cellular targets. To overcome this, it has been suggested that they be modified chemically. Among the various options, conjugation with short- and long-chain polyethylene glycols (PEGs) has several advantages: PEG is nontoxic and very soluble, reduces immunogenic reactions, and increases the stability of the conjugated molecules. PEG is generally joined to oligonucleotides while bound to the insoluble solid-phase supports, after their synthesis, which does not allow for their being easy scaled up. The use of the liquid-phase approach as an alternative synthetic process, utilizing the PEG polymer both as a soluble, inert synthetic support and a stabilizing agent of the oligonucleotide, is proposed. A new protocol has been set up, characterized by a stable phosphate bond between the support and the oligonucleotide. This method has been tested on a 12mer previously investigated as an antisense agent against HIV. The PEG-conjugated 12mer was efficiently synthesized and purified. It was found that the high-molecular mass PEG chain results in enzymatic stability and does not interfere with the formation of the duplex with its nucleic acid target. PMID- 9404653 TI - Studies on base-boronated oligonucleotides. 2 (1). Incompatibility of DMT and cyanoborane groups during oligonucleotide synthesis. AB - The cyanoborane (-BH2CN) nucleosides and nucleotides are a new class of compounds that mimic natural and synthetic congeners in many ways and exhibit interesting biochemical and biophysical properties. The B-N bond is isoelectronic with the C N+ bond of N7-alkylated 2'-nucleosides, as well as the C-C bond of naturally occurring 7-alkyl-7-deazanucleosides. These compounds differ from normal guanosine in that they are incapable of hydrogen bonding at the 7-position. The syntheses of N7-cyanoborane 2'-deoxyguanosine, N2-(dimethylaminomethylene)-N7 cyanoborane 5'-(dimethoxytrityl)-2'-deoxyguanosine (3), and N2-isobutyryl-N7 cyanoborane 5'-(dimethoxytrityl)-2'-deoxyguanosine (9) are described. Removal of the dimethoxytrityl (DMT) group from 3 or 9 is accompanied by significant loss of the cyanoborane moiety. Additionally, dimethoxytritylation of a cyanoboronated nucleoside leads to partial deboronation, thus limiting use of the commercially available 5'-DMT nucleosides as viable precursors in base-boronated oligonucleotide synthesis. The incompatibility of the cyanoborane moiety under DMT removal/addition conditions necessitated the search for an alternative method of protecting the 5'-hydroxyl of the nucleoside. This paper addresses the possible cause of deboronation and describes the synthesis of N7-cyanoboronated nucleosides by a method that avoids transient protection of the sugar hydroxyls. PMID- 9404652 TI - Role of the central metal ion and ligand charge in the DNA binding and modification by metallosalen complexes. AB - Several metal complexes of three different functionalized salen derivatives have been synthesized. The salens differ in terms of the electrostatic character and the location of the charges. The interactions of such complexes with DNA were first investigated in detail by UV-vis absorption titrimetry. It appears that the DNA binding by most of these compounds is primarily due to a combination of electrostatic and other modes of interactions. The melting temperatures of DNA in the presence of various metal complexes were higher than that of the pure DNA. The presence of additional charge on the central metal ion core in the complex, however, alters the nature of binding. Bis-cationic salen complexes containing central Ni(II) or Mn(III) were found to induce DNA strand scission, especially in the presence of co-oxidant as revealed by plasmid DNA cleavage assay and also on the basis of the autoradiogram obtained from their respective high-resolution sequencing gels. Modest base selectivity was observed in the DNA cleavage reactions. Comparisons of the linearized and supercoiled forms of DNA in the metal complex-mediated cleavage reactions reveal that the supercoiled forms are more susceptible to DNA scission. Under suitable conditions, the DNA cleavage reactions can be induced either by preformed metal complexes or by in situ complexation of the ligand in the presence of the appropriate metal ion. Also revealed was the fact that the analogous complexes containing Cu(II) or Cr(III) did not effect any DNA strand scission under comparable conditions. Salens with pendant negative charges on either side of the precursor salicylaldehyde or ethylenediamine fragments did not bind with DNA. Similarly, metallosalen complexes with net anionic character also failed to induce any DNA modification activities. PMID- 9404654 TI - Biotin reagents for antibody pretargeting. 2. Synthesis and in vitro evaluation of biotin dimers and trimers for cross-linking of streptavidin. AB - Polymerization and/or cross-linking of recombinant streptavidin (r-SAv) with biotin derivatives containing two biotin moieties (biotin dimers) or three biotin moieties (biotin trimers) has been investigated as a model for reagents to be used to increase the amount of radioactivity on cancer cells in tumor pretargeting protocols. In the investigation, six biotin dimers and three biotin trimers were synthesized. Most biotin derivatives synthesized had ether containing linker molecules incorporated to improve their aqueous solubility. The synthesized biotin dimers contained linker moieties which provided distances (when fully extended) of 13-49 A between biotin carboxylate carbon atoms, and the biotin trimers contained linker moieties which provided distances of 31-53 A between any two biotin carboxylate atoms. All of the biotin derivatives were evaluated for their ability to polymerize r-SAv in solution. When the biotin derivatives were mixed with r-SAv, none of the biotin dimers caused polymerization, but all of the biotin trimers resulted in complete polymerization. Some of the biotin dimers did cross-link r-SAv (to form r-SAv dimers, trimers, etc.), but the percentage of cross-linking was low (< or = 40%). The length of the linker molecule was important in cross-linking of biotin dimers. While linkers which provided distances of 13 and 19 A between biotin carboxylate carbon atoms did not result in cross-linking, a linker which provided a 17 A distance resulted in a small (< or = 10%) amount of cross-linking. Also cross-linking was increased in biotin dimers with linkers which provided distances between biotin carboxylate carbon atoms of > or = 23 A. Cross-linking of streptavidin bound in polystyrene wells with biotin dimers and trimers was also examined. In those experiments, an excess of each biotin derivative was incubated at 37 degrees C for 10-30 min in polystyrene wells containing bound SAv. After the excess biotin derivative was rinsed from the wells, an excess of r [125I]SAv was incubated for another 10-30 min. The amount of r-[125I]SAv bound after rinsing the excess from the wells was an indicator of the extent of cross linking that occurred. The process of alternating additions of reagents was repeated four times to demonstrate that bound radioactivity could be increased with each addition of [125I]SAv. The results of cross-linking r-SAv in polystyrene wells paralleled results from cross-linking in solution. PMID- 9404655 TI - Design of comb-type polyamine copolymers for a novel pH-sensitive DNA carrier. AB - The comb-type polycation consisting of a poly[2-(diethylamino)ethyl methacrylate] (PDEAEMA) backbone and poly(L-lysine) (PLL) side chains has been prepared as a novel pH-sensitive DNA carrier. The comb-type copolymer PDEAEMA-graft-PLL was prepared by using the macromonomer method, in which a poly(N epsilon-carbobenzoxy L-lysine) macromonomer was radically copolymerized with DEAEMA. The comb-type copolymer exhibited a two-step proton dissociation and a dual ionic character owing to the two cationic segments in the copolymer, as determined by acid-base titration. In addition, the comb-type copolymer caused no significant turbidity even at pH 10, whereas PDEAEMA homopolymer suddenly precipitated out of the aqueous medium above pH 7.5 owing to the deprotonation of amino groups. Furthermore, a 1H NMR study proved that protonated PLL segments solubilized the comb-type copolymer with a hydrophobic PDEAEMA core at higher pH. Finally, the pH dependent behavior of the DNA complex with the comb-type copolymer was evaluated. The discontinuous turbidity change of the DNA/PDEAEMA-graft-PLL mixture at pH 7.5 suggested that the solubility of the complex varied in response to pH. By circular dichroism measurement, we also found that the comb-type copolymer was capable of varying DNA compaction pH-dependently. In conclusion, we have demonstrated that the comb-type copolymer is capable of sensing a pH signal and outputting the nonlinear change of the physicochemical properties of DNA polyelectrolyte complexes. PMID- 9404656 TI - In vitro gene delivery to hepatocytes with galactosylated polyethylenimine. AB - A hepatocyte-directed vector has been developed; it includes several key features thought to favor in vivo gene delivery to the liver: electrostatically neutral particles which avoid nonspecific binding to other cells, to the extracellular matrix, and to complement proteins; asialoglycoprotein receptor-mediated endocytosis which may address the complexes to the perinuclear region; and polyethylenimine (PEI)-mediated endosome buffering and swelling as an escape mechanism to the cytoplasm. This system is based on a 5% galactose-bearing polyethylenimine (PEI-gal) polymer which is condensed with plasmid DNA to neutrality. Murine (BNL CL.2) and human (HepG2) hepatocyte-derived cell lines were transfected 10(4)-10(5)-fold more efficiently than murine fibroblasts (3T3), whether transfection was assessed globally (luciferase expression from the cell extract) or following histochemical staining (beta-galactosidase). Under these conditions, over 50% of the hepatocytes were selectively transfected in the presence of 10% serum. Transfection was suppressed by removal of the targeting galactose residues, by their replacement with glucose, or by the addition of excess asialofetuin. Thus, results from comparative and competitive experiments indicate the asialoglycoprotein receptor is involved in transfection of hepatocytes with neutral PEI-gal/DNA complexes. PMID- 9404657 TI - Neoglycoproteins with the synthetic complex biantennary nonasaccharide or its alpha 2,3/alpha 2,6-sialylated derivatives: their preparation, assessment of their ligand properties for purified lectins, for tumor cells in vitro, and in tissue sections, and their biodistribution in tumor-bearing mice. AB - Neoglycoproteins were prepared with chemoenzymatically synthesized complex biantennary N-glycan derivatives the nonreducing ends of which bear typical sequences found in glycoproteins. A chemically obtained biantennary heptasaccharide-azide was reduced and acylated with a 6-aminohexanoyl spacer. Elongation of the deprotected heptasaccharide using glycosyltransferases yielded a biantennary nonasaccharide with terminal galactose residues and two undecasaccharides terminating with alpha 2,6- or alpha 2,3-linked sialic acid. The free amino group of the spacer of these oligosaccharides was converted into an isothiocyanate. Its subsequent coupling to bovine serum albumin gave neoglycoproteins with a yield of 2.4-3.6 glycan chains per carrier molecule. This versatile synthetic pathway allows employment of a wide variety of complex-type glycans, which can be introduced to various test systems in vitro and in vivo to evaluate potential biomedical applications. Solid-phase assays with biotinylated sugar receptors revealed discriminatory binding properties of the three neoglycoproteins, especially for the mistletoe lectin. This direct assay system is preferable to the measurement of inhibitory capacities with respect to model ligands. Ligand type- and cell type-dependent quantitative differences in the binding properties of the probes were detected by FACScan analyses with a panel of tumor cell lines and by monitoring of staining in tissue sections for small cell and non-small-cell lung cancer and mesotheliomas. Biodistribution of iodinated neoglycoproteins in mice gave a prolonged presence of the sialylated probes in serum. Relative to the nonasaccharide, the uptake, especially of the iodinated neoglycoprotein with alpha 2,3-sialylated ligand chains, was clearly elevated in mice for kidneys and Ehrlich tumors. On the basis of the documented feasibility of these applications, it is concluded that the further elaboration of glycan chain variants by the described synthetic approach in combination with the given test panel is warranted to evaluate the potential of complex glycan chain-carrying neoglycoproteins for diagnostic and therapeutic purposes. PMID- 9404658 TI - Esterase-triggered fluorescence of fluorogenic oligonucleotides. AB - In the prooligonucleotide approach, a step of activation by cellular esterases is necessary for the removal of internucleoside phosphate masking groups and subsequent intracellular delivery of active antisense oligonucleotides. The efficacy of this approach implies that prooligonucleotides, once they are taken up by cells, are demasked by esterases during their course to their nucleic acid targets. In this regard, a method for labeling oligomers with esterase-activable fluorogenic tag was designed. The two phenolic functions of carboxyfluorescein were protected by pivaloyl groups, yielding a nonfluorescent lactone which was further activated as a N-hydroxysuccinimide ester. Two nuclease-resistant phosphorothioate 18-mer and methylphosphonate 19-mer oligodeoxynucleosides were attached to this biprotected fluorescein derivative via an amino linker at the 5' end of the oligomers. The two conjugates were assayed for their carboxyesterase substrate ability in different biological media. In the presence of purified esterases or when incubated in serum or cell extracts, both oligonucleotide conjugates became fluorescent. In addition, the phosphorothioate oligoconjugate was microinjected into the cytoplasm of human fibroblasts, and a fast cytoplasmic release of fluorescence was observed with a rapid translocation of the fluorescent oligomer into the nucleus. PMID- 9404659 TI - Incorporation of an artificial receptor into a native protein: new strategy for the design of semisynthetic enzymes with allosteric properties. AB - The sugar-facilitated structure and enzymatic activity change of engineered myoglobins bearing a phenylboronic acid moiety, which were semisynthesized by a cofactor reconstitution method, were studied by the denaturation experiment, spectrophotometric titration of the pKa shift of the axial H2O, circular dichloism (CD), and the kinetics of the myoglobin-catalyzed-aniline hydroxylation reaction. Both boronophenylalanine-appended myoglobin [Mb(m-Bphe)2] and phenylboronic acid-appended myoglobin [Mb(PhBOH)2] were stabilized by approximately 2 kcal/mol upon complexation with D-fructose. CD spectral changes and the sugar-induced pKa shift suggested that the microenvironment of the active site of these myoglobins was re-formed from a partially disturbed state to that comparable to the native state upon D-fructose binding. The correlation of pKa with kcat (for the aniline hydroxylase activity) and the delta GDH2O-kcat profile showed that these structural changes of Mb-(m-Bphe)2 and Mb(PhBOH)2 were closely related to their sugar-enhanced aniline hydroxylase activity. Thus, the results established that an incorporation of the artificial receptor molecule can be a valid methodology for the design of stimuli-responsive semiartificial enzymes. PMID- 9404660 TI - Bisintercalation of homodimeric thiazole orange dyes in DNA: effect of modifying the linker. AB - The thiazole orange dye 1,1'-(4,4,8,8-tetramethyl-4,8-diazaundecamethylene)-bis[4 [3-methy l-2, 3-dihydro(benzo-1,3-thiazole)-2-methylidene]]quinolinium tetraiodide (TOTO) binds to double-stranded DNA (dsDNA) in a sequence selective bisintercalation. Each chromophore is sandwiched between two base pairs in a (5' CpT-3'):(5'-ApG-3') site, and the linker spans over two base pairs in the minor groove. The binding of analogs of TOTO in which the linker has been modified is examined. The aim of the study is to utilize the sequence selectivity of the TOTO chromophores to enhance and/or alter the overall selectivity of the binding. One- and two-dimensional 1H-NMR investigations of complexes between TOTO analogs and various dsDNA oligonucleotides are reported. The following analogs were synthesized and used: 1,1'-(4,4,8,8-tetramethyl-4,8-diazadodecamethylene) -bis[4 [3-methyl-2,3-dihydro- (benzo-1,3-thiazole)-2-methylidene]]quinolinium tetraiodide (TOTO10), 1,1'-(5,5,9,9-tetramethyl-5,9-diazatridecamethylene)-bis[4 [3-meth yl-2, 3-dihydro(benzo-1,3-thiazole)-2-methylidene]]quinolinium tetraiodide (TOTO11), and 1,1'-(6,6,10,10-tetramethyl-6,10 diazapentadecamethylene)-bis[4-[3 -methyl-2, 3-dihydro(benzo-1,3-thiazole)-2 methylidene]]quinolinium tetraiodide (TOTO13). The results show that with a longer linker the dyes can bisintercalate into two (5'-CpT-3'):(5'-ApG-3') sites separated by one or two base pairs. Bisintercalation in two such "isolated" binding sites yields non-nearest-neighbor bisintercalation in which the linker spans over more than two base pairs. The investigations also showed that an exact length of the linker is not crucial for the site selectivity since TOTO, TOTO10, and TOTO11 are almost equally suitable in binding selectively to the (5'CTAG-3')2 sequence. Fluorescence measurements show that TOTO10, TOTO11, and TOTO13 have higher fluorescence quantum yields than TOTO when bound to d(CGCTAGCG)2. This indicates that the length of the linker in TOTO may not be the optimum one in terms of using the dye as a fluorescence marker. PMID- 9404662 TI - Libraries of multifunctional RNA conjugates for the selection of new RNA catalysts. AB - An in vitro selection system was developed for the selection of RNA molecules catalyzing bimolecular reactions between small reactants. The system is based on the direct selection protocol and involves libraries of multifunctional RNA conjugates rather than unmodified RNA transcripts. For the preparation of RNA conjugate libraries, a dinucleotide analog has been designed and synthesized containing a poly(ethylene glycol) linker with an embedded photocleavage site and a terminal attachment site for coupling potential reactants. Reactants are first coupled to the dinucleotide analog by activated ester chemistry and then ligated to the 3'-ends of enzymatically prepared RNA pool molecules, giving libraries of complex conjugates. Species that become attached to biotin on incubation with a biotinylated partner are isolated using streptavidin-derivatized matrices and then subjected to a photocleavage step. Selective cleavage of the linker releases only those RNA species in which reaction has taken place at the linker-coupled reactant, while products with the biotin attached to internal positions of the RNA part remain immobilized. Efficient photocleavage is achieved by laser irradiation at 355 nm, and the released RNAs are intact and amplifiable by reverse transcription. All steps are shown to be compatible with the overall selection procedure, as was shown by performing a model selection cycle. Besides allowing a broader scope of reaction types to be selected for, the strategy relieves the RNA from the requirement to possess substrate properties as well as catalytic activity, and the use of a cleavable linker will suppress the selection of catalysts for side reactions. PMID- 9404661 TI - Generation and characterization of an anti-CD19 single-chain Fv immunotoxin composed of C-terminal disulfide-linked dgRTA. AB - Our laboratory utilized two methods to produce the anti-CD19 immunotoxin containing a single-chain Fv (scFv) FVS191 and a ricin A chain (RTA). The first method produced the recombinant protein FVS191CDRTA from a fusing gene containing sequences encoding FVS191, catheptsin D proteinase digestion site (CD), and RTA. FVS191CDRTA did not show CD19 antigen binding and cytotoxic activity. The second method generated a disulfide-linked FVS191cys-dgRTA from a FVS191cys, the FVS191 with an additional C-terminal cysteine, and a deglycosylated RTA (dgRTA). The formation of FVS191cys-dgRTA is efficient; up to 70% of the proteins participating in the reaction had formed FVS191cys-dgRTA when the molar ratio of FVS191cys to dgRTA was 1:1. A competitive ELISA assay indicated that FVS191cys dgRTA and the parental monoclonal antibody B43 possessed comparable CD19 binding abilities. The protein synthesis inhibition assay revealed that FVS191cys-dgRTA was toxic to CD19 positive cell lines, but it was less potent than the intact antibody-conjugated B43-dgRTA, which had an IC50 = 2 x 10(-11) M. 125I-Labeled FVS191 and 125I-labeled B43 were internalized by Nalm-6 cells at 37 degrees C as demonstrated by internalization studies; this result indicates that cross-linking of CD19 antigen is not required for the endocytosis of CD19 and raises the possibility that the lower cytotoxity of FVS191cys-dgRTA is not due to the monovalent binding of CD19 by FVS191cys-dgRTA. Our study with anti-CD19 scFv immunotoxin indicates that the formation of a disulfide-linked scFv immunotoxin is an alternative to the recombinant method of producing scFv immunotoxin. PMID- 9404663 TI - Evidence for highly cooperative binding between molecular umbrella-spermine conjugates and DNA. AB - Double- and tetrawalled molecular umbrella-spermine conjugates (I and II) have been synthesized, and their binding to calf thymus DNA (CT-DNA), poly[d(AT)], and poly[d(GC)] compared with that of a single-walled analogue (III). At moderate salt concentrations (8 mM NaCl), I and II show significantly greater affinity toward each DNA, relative to III; at high salt concentrations (150 mM NaCl), strong binding of I and II (but not III) was maintained toward poly[d(GC)]. Examination of the influence of I-III on the melting behavior of poly[d(AT)] has provided strong evidence that the binding of I and II reflects highly cooperative interactions among DNA-bound conjugates and that the DNA duplex serves as a nucleation site for umbrella aggregation. The implications of these findings for the rational design of novel drug conjugates that operate at the nuclear level, and also novel transfection agents, are briefly discussed. PMID- 9404664 TI - New synthesis and characterization of (+)-lysergic acid diethylamide (LSD) derivatives and the development of a microparticle-based immunoassay for the detection of LSD and its metabolites. AB - In this paper are reported the synthesis and characterization of three LSD derivatives. On the basis of several analytical characterization studies, the most stable derivative has been selected and a procedure to covalently link the derivative to polystyrene microparticles through a carrier protein has been developed. In addition, two new LSD immunogens have been synthesized and characterized, and from these immunogens antibodies that recognize not only LSD but also several major LSD metabolites have been generated. Using the selected derivative and antibody, a homogeneous microparticle-based immunoassay has been developed for the detection of LSD in human urine with the required sensitivity and specificity for an effective screening assay. The performance of this LSD OnLine assay has been evaluated using the criteria of precision, cross reactivity, correlation to the Abuscreen LSD RIA and GC/MS/MS, assay specificity, and limit of detection. PMID- 9404665 TI - Probing biomolecule recognition with electron transfer: electrochemical sensors for DNA hybridization. AB - Identifying infectious organisms, quantitating gene expression, and sequencing genomic DNA on chips all rely on the detection of nucleic acid hybridization. Described here is a novel assay for detection of the hybridization of products of the polymerase chain reaction using electron transfer from guanine to a transition-metal complex. The hybridization assay was modeled in solution by monitoring the cyclic voltammetry of Ru(bpy)3(2+) (bpy = 2,2'-bipyridine) in the presence of a probe strand containing only A, T, and C prior to and after hybridization to a complement that contained seven guanines, which led to high catalytic current due to the oxidation of guanine by Ru(bpy)3(3+). To allow recognition of all four bases in the target sequence, it was shown that inosine 5'-monophosphate was 3 orders of magnitude less reactive than guanosine 5' monophosphate, suggesting that effective hybridization sensors could be realized by immobilization of probe strands in which inosine was substituted for guanosine; hybridization to guanosine-containing target strands would then provide high catalytic currents. A sensor design was tested in a model system for the detection of a synthetic 21-mer oligonucleotide patterned on the sequence of the ras oncogene, which gave an increase in charge collected of 35 +/- 5 microC after hybridization and of only 8 +/- 5 microC after exposure to noncomplementary DNA. Independent quantitation of probe and target by radiolabeling showed that the hybridized electrode contained 3.0 +/- 0.3 ng of target. New sensor electrodes were then prepared for the detection of PCR-amplified genomic DNA from herpes simplex virus type II, genomic DNA from Clostridium perfringens, and genomic RNA from human immunodeficiency virus and gave an additional charge of 35 65 microC for hybridization to complementary amplicon and of only 2-10 microC after exposure to noncomplementary DNA. PMID- 9404666 TI - Development of a kinetic model to describe the effective rate of antibody oxidation by periodate. AB - The oxidation of antibody carbohydrate residues by periodate is a common approach used for site-specific antibody modification and immobilization. This study sought to develop a general kinetic model that could be used to describe the effective rate of this oxidation for process control. A detailed analysis of previous data collected for rabbit immunoglobulin G in the presence of excess periodate indicated that the reaction followed a pseudo-first-order mechanism in which two general classes of sites were being oxidized. The first class of sites was oxidized fairly rapidly (i.e., within 15-30 min), while the second class of sites reacted over the course of several hours. From these results, an equation was developed that gave a good fit under a variety of reaction conditions to the production of oxidized sites available for coupling with a hydrazide label. On the basis of this equation, data obtained at several periodate concentrations under the same pH and temperature conditions were used to estimate the apparent rate and equilibrium constants for the oxidation of each class of sites. The values obtained by using this approach could be used not only to predict the effective rate of oxidation at other periodate concentrations but also to provide information on the individual steps involved in the oxidation process. PMID- 9404667 TI - Efficient chemical introduction of a disulfide cross-link and conjugation site into human hemoglobin at beta-lysine-82 utilizing a bifunctional aminoacyl phosphate. AB - The creation of a cross-link containing a disulfide into hemoglobin has been accomplished with a site-directed reagent, N,N'-bis(Cbz-cystinyl)bis(methyl phosphate) (1). This is prepared from the reaction of the bis acid chloride of N protected cystine with dimethyl phosphate followed by O-demethylation with methyl iodide in acetone. Reaction with deoxyhemoglobin produces two main products: cross-linked hemoglobin as the bis(cystinyl amide) of the epsilon-amino group of the side chain of Lys-82 of the two beta subunits as well as material that has each of the same amino groups modified as the cysteinyl amide but not cross linked. Addition of 2-mercaptoethanol cleaves the disulfide in the material that is not cross-linked while leaving the disulfide intact in the cross-linked species. Dithiothreitol reduces the disulfide in the cross-linked species as well as in the species that is not cross-linked. Spontaneous oxidation in air converts all of the reduced material to the cross-linked bis(cystinyl amide) of hemoglobin. The reagent permits controlled introduction of cystinyl groups at lysyl residues, leading to formation of sulfhydryl groups by reduction and the possibility of re-forming the cross-links or forming conjugates. PMID- 9404668 TI - Preparation and purification of an end to end coupled mEGF-dextran conjugate. AB - The amino terminus of mouse epidermal growth factor (mEGF) was coupled directly to the aldehyde end of dextran through a reductive amination procedure. The highest coupling efficiency was approximately 80% and could be reached after approximately 24 h of reaction time at pH 8. Gel filtration on Sephadex G-50 Fine removed free mEGF from the conjugate. Preparative polyacrylamide gel electrophoresis was used to separate the conjugate from excess noncharged dextran. The conjugate bound specifically to the EGF receptor on cultured glioma cells as shown in displacement tests with free mEGF. The conjugate was stable in the pH interval 4-9, in 2 M sodium chloride, in 7 M urea, and in human serum and could still bind to the EGF receptor after such treatments. The conjugates are candidates for targeted nuclide therapy. PMID- 9404670 TI - Bilayer distribution of phosphatidylserine and phosphatidylethanolamine in lipid vesicles. AB - The distribution of phosphatidylethanolamine (PE) and phosphatydilserine (PS) in liposomes was studied as a function of aminophospholipid concentration using fluorescamine as labeling reagent. The method is suitable for such determination since, in the assay conditions, fluorescamine does not penetrate the vesicles nor does it disrupt them. The liposomes were obtained by sonication, extrusion, or mechanical dispersion (MLV). For any kind of vesicle, the percentage of PS in the external monolayer is higher than that obtained for PE in the corresponding vesicles. In extruded PS liposomes, this aminophospholipid is located preferentially in the outer layer, while for PE liposomes the localization depends on the size of vesicle. Sonicated liposomes present an asymmetrical distribution of both aminophospholipids, and the external location of PS or PE always predominates. In contrast, in MLV, aminophospholipids are mainly found in the inner layers of the vesicles, except for liposomes formed by the lowest PS proportion. A remarkable feature of PS liposomes is the reduction of vesicle size, especially in MLV liposomes, in comparison with neutral liposomes. PMID- 9404669 TI - Covalent protein-oligonucleotide conjugates for efficient delivery of antisense molecules. AB - Antisense oligonucleotides have been covalently attached to asialoglycoprotein (ASGP) via disulfide bond conjugation chemistry. These conjugates were characterized extensively by an array of chemical, chromatographic, and spectroscopic means. Multiple (approximately six) oligonucleotides can be conjugated to each ASGP molecule. The molecular conjugates were used to deliver antisense oligonucleotides complementary to the mRNA of the interleukin 6 signal transduction protein (gp130) to modulate the acute phase response of hepatoma (HepG2) cells in vitro. These conjugates were biologically active, as measured by inhibition of the cytokine-stimulated up-regulation of the acute phase protein haptoglobin. The level of inhibition was comparable to that found with previous technology featuring noncovalent complexes of ASGP-poly(L-lysine) and oligonucleotide. Because of the ability to control the stoichiometry of the conjugate and its unimolecular nature (as opposed to bimolecular for the noncovalent conjugates), this methodology holds great promise for further development and application. PMID- 9404671 TI - Some observations on the trace element concentrations in human dental enamel. AB - The concentration of trace elements has been measured for dental enamel from 86 healthy human teeth using particle-induced X-ray emission (PIXE). The majority of the teeth (n = 70) were collected from dentists in the county of Oxfordshire in the United Kingdom, although a smaller group (n = 16) were collected from Cornwall. The elements K, Ca, Mn, Fe, Co, Ni, Cu, Zn, Sr, Pb, and Hg have been detected and statistically analyzed by grouping according to sex, age, and geographical location. The concentrations of Fe and Cu were found to be lower in the teeth from female donors (P < 5%) and are believed to result from the continued burden of blood loss during menstruation. Strong positive correlations (P < 0.1%) were found between Ca, Co, Ni, and Zn for all groups; these elements were also found to exhibit a negative correlation (P < 1%) with age for teeth from female donors. This is believed to be related to decalcification during the menopause. Pb was found to exhibit a positive correlation (P < 5%) with age for both sexes, and is believed to substitute for Ca in the Ca hydroxy apatite (HAP) within the dental enamel. PMID- 9404672 TI - Lead disrupts eicosanoid metabolism, macrophage function, and disease resistance in birds. AB - Lead (Pb) affects elements of humoral and cell-mediated immunity, and diminishes host resistance to infectious disease. Evidence is presented supporting a hypothesis of Pb-induced immunosuppression stemming from altered fatty acid metabolism, and mediated by eicosanoids and macrophages (MO). Chronic Pb exposure increases the proportion of arachidonate (ArA) among fatty acids in lipid from avian tissues, and this change provides precursors for eicosanoids, the oxygenated derivatives of ArA that mediate MO acute inflammatory response. In the current study, we showed that the concentration of ArA in phospholipids of MO elicited from turkey poults fed 100 ppm dietary Pb acetate was twice that of controls. In vitro production of eicosanoids by these MO was substantially increased, and this effect was most pronounced following lipopolysaccharide stimulation: prostaglandin F2 alpha was increased 11-fold, thromboxane B2 increased threefold, and prostaglandin E2 increased by 1.5 times. In vitro phagocytic potential of these MO was suppressed, such that the percentage of MO engulfing sheep red blood cell (RBC) targets was reduced to half that of control MO. In vivo susceptibility of Pb-treated and control birds to Gram-negative bacteria challenge was also evaluated. The morbidity of chicks inoculated with Salmonella gallinarum and fed either control or 200 ppm Pb acetate-supplemented diets was similar, except early in the course of the disease when mortality among Pb-treated birds was marginally greater. In these studies, effects of Pb that could influence immunological homeostasis were demonstrated for MO metabolism of ArA, for production of eicosanoids, and for phagocytosis. There was also the suggestion that these in vitro indices of immune function are related to in vivo disease resistance. PMID- 9404673 TI - Determination of trace boron in microsamples of biological tissues. AB - A benign-by-design method for the determination of boron (B) in microsamples of biological tissues was developed. This is a simple, automated, microdigestion method. Use of reagents and generation of waste are minimized, and the use of toxic/hazardous reagents is eliminated as compared to currently available B methodology. Microsamples are accommodated by the method; 100-400 mg samples were used in this study. B is determined by inductively coupled plasma atomic emission spectrometry (ICPAES) at 249.678 nm. The instrument detection limit for B is 0.01 microgram/mL. Interference studies have been investigated for 21 common elements. Over 250 analyses of standard reference materials were analyzed during the study duration. Recoveries for a series of biological tissues, both plant and animal, ranged from 82-104%. PMID- 9404674 TI - Disturbances of the mineral incorporation in various species of mice and shrews in the emission area of a phosphate plant. AB - The Cd emission of a phosphate plant was clearly reflected by the Cd status of herbivorous European wood mice and common field voles as well as of European shrews taking in mostly animal food. The antagonistic effect of the emitted Cd and Mo better available for plants with high ground pH most probably caused the deterioration in the Cu status of the animals of both phases in the nutritional chain. The lower Ca, P, and Mg incorporation with European wood mouse and common field vole within the contaminated habitat might as well be owing to emission, whereas the lower Mn content in all three species rather has to be attributed to the lower Mn offer caused by the ground pH. PMID- 9404675 TI - Supplementary selenium influences the response to fatty acid-induced oxidative stress in humans. AB - The mutual influences of wheat selenium (Se) and n-3 polyunsaturated fatty acids (n-3 PUFA) on plasma Se and indicators of increased oxidative stress were investigated in a randomized, double-blind study with 31 women (23.5 +/- 3.4 yr). Groups 1 and 2 ingested 5.4 g n-3 PUFA daily (as ethyl esters), whereas groups 3 and 4 received placebo capsules. Groups 2 and 3 received 3 slices of high-Se bread daily, providing 115 micrograms Se, in addition to the 77 +/- 26 micrograms Se in the diet. Groups 1 and 4 received placebo slices. Blood samples were drawn at baseline and at 3 and 6 wk. Serum Se concentrations increased in both groups given Se-enriched bread, but significantly less in subjects given n-3 PUFA (group 2). There were no changes in the plasma ratio alpha-tocopherol:mg cholesterol or plasma ascorbic acid levels. In group 1, plasma-conjugated dienes and thiobarbituric acid-reactive substances (TBARS) rose by 130% (p < 0.005) and 126% (p < 0.005), respectively. Two-way ANOVA showed significant interaction effects of Se and n-3 PUFA on changes in conjugated dienes (p = 0.03) and TBARS (p = 0.015), Se treatment apparently modifying the peroxidative effects of n-3 PUFA. In subjects receiving n-3 PUFA, changes in conjugated dienes and TBARS were negatively correlated with changes in serum Se. In summary, n-3 PUFA modified the effect of Se supplementation, whereas Se seemed to modify the peroxidative effects of n-3 PUFA. PMID- 9404676 TI - A fluorescence double-quenching study of native lipoproteins in an animal model of manganese deficiency. AB - Iodide and acrylamide were applied simultaneously in a double-quenching experiment to compare acrylamide quenching constants for internal and external fluorophores of high-density lipoproteins (HDL1 and HDL2) from manganese-adequate (MnA) and deficient (MnD) rats, free of the electrostatic effects associated with iodide. In MnA HDL1 compared to MnD HDL1, the acrylamide quenching constant for external fluorophores was different (P < 0.1). In MnA HDL2, there were two populations of fluorophores accessible to acrylamide, whereas in MnD HDL2, all fluorophores were accessible to both quenchers. We concluded that there were structural (local environmental) differences, possibly charge-related, around the external fluorophores, and a slightly larger population of buried fluorophores in the MnD HDL1 compared with MnA HDL1. In MnA HDL2, one-third of the fluorophores were accessible to iodide, and all external and internal fluorophores were accessible to acrylamide, whereas in MnD HDL2, all fluorophores were accessible to both quenchers. PMID- 9404677 TI - Age-dependent changes of mineral contents in men and women's calcanei. AB - To elucidate age-related change of mineral contents in human bones, the mineral content and density of human calcanei were determined by inductively coupled plasma atomic emission spectrometry and dual-energy X-ray absorptiometry. Calcanei were removed from 27 subjects (17 men and 10 women) who died in the age range from 40-98 yr old. Both the inductively coupled plasma emission spectrometry and dual-energy X-ray absorptiometry indicated that there were age dependent decreases of the mineral contents and density in the men's calcaneus in the age range from 40-98 yr, but not in the women's calcaneus in the age range from 42-87 yr. It was also found that the calcanean masses of the men and women remained constant within the same age range until 98 yr. PMID- 9404678 TI - An elemental correlation study in cancerous breast tissue by total reflection x ray fluorescence. AB - The total reflection x-ray fluorescence method (TRXRF) has been employed to determine of P, S, K, Ca, Cr, Mn, Fe, Ni, Cu, Zn, Se, Rb, Sr, and Pb concentration in the benign breast tumor tissue from 68 women and in the cancerous breast tissue from 26 women. Concentrations of most of elements show enhancement in cancerous breast tissue. Examined elements compete for binding sites in the cell, change its enzymatic activity, and exert direct or indirect action on the carcinogenic process accelerating the growth of tumors. Inhibition of enzymatic activity caused by variation in trace element concentrations results in immunological breakdown of the body system. An attempt has been made to correlate measured trace element concentrations with the clinical stage of cancer. Physical bases of used analytical method, experimental setup, and the procedure of sample preparation are described. PMID- 9404679 TI - Daily zinc supplementation effect on zinc deficiency in rats during prolonged restriction of motor activity. AB - The objective of this investigation was to evaluate the effect of 47 mg zinc supplementation on deficiency of zinc in rats during 98 d of restriction of motor activity (hypokinesia), which appeared by higher plasma zinc concentration. One Hundred 13-week-old Sprague-Dawley male rats weighing 360-390 g were used to perform the studies: They were equally divided into four groups: 1. Unsupplemented control animals (UCA); 2. Unsupplemented hypokinetic animals (UHA); 3. Supplemented control animals (SCA); and 4. Supplemented hypokinetic animals (SHA). For the simulation of the effect of hypokinesia (HK), the UHA and SHA were kept in small individual cages made of wood, which restricted their movements in all directions without hindering food and water intake. The SCA and SHA received daily with their food an additional amount of zinc. Before and during the experimental period of 98 d, plasma, urinary and fecal zinc, balance of zinc, food intake, and body weight were determined at different intervals. In the SHA and UHA, the concentration of zinc in plasma, and the elimination of zinc in urine and feces increased significantly when compared with the SCA and UCA, whereas the balance of zinc was negative. The body weight and food intake decreased significantly in the SHA and UHA when compared with the SCA and UCA. The increased plasma concentration of zinc in both the SHA and UHA groups was in contrast to the observed hypozincnemia during prolonged immobilization as during prolonged hospitalization. This reaction suggests that there may be some other mechanisms that are affecting the process of control and regulation of zinc metabolism during prolonged HK. It was concluded that exposure to prolonged restriction of motor activity of rats induces significant increases in plasma concentration, fecal and urinary elimination of zinc in the presence of negative zinc balance and regardless the daily intake of large amounts of zinc with their food, leading to zinc deficiency. PMID- 9404680 TI - Microcalorimetric study of the toxic effect of selenium on the mitochondrial metabolism of Cyprinus carpio liver. AB - The metabolic thermograms and heat output of mitochondria isolated from carp liver have been determined by using an LKB bioactivity monitor. The thermogram can be divided into four parts: the lag phase, active recovery phase, stationary phase, and decline phase. The thermokinetic equation was established for the active recovery and decline phase of metabolism as follows: dP/dt = k(m)P (1-SP). The rate constants k1 and k2 of two phases of active recovery and decline phase have been also calculated. The metabolism activity of mitochondrial inhibited by a high concentration of trace element selenium has been studied. The metabolic heat released, time of each phase, and rate constants can be significantly influenced by excess of selenite added. These results suggested that a high concentration of selenium can damage the structure and function of mitochondria, and thus influence their metabolism. PMID- 9404681 TI - Zinc status in plasma of obese individuals during glucose administration. AB - To know whether plasma zinc status is altered under acute hyperglycemic state, the interrelationships among plasma glucose, insulin, and zinc concentrations during oral glucose tolerance test (OGTT) in obese individuals and their lean controls were studied. Plasma glucose and insulin concentrations under fasting as well as those values in response to OGTT were significantly higher in obese individuals than those in lean controls. On the other hand, the obese had lower fasting plasma zinc concentrations compared to lean controls (13.5 vs 18.1 mumol/L, p < 0.005). Under fasting, plasma zinc concentrations in overall individuals inversely correlated to their body mass index (BMI) (r = -0.516), plasma glucose (r = -0.620), and plasma insulin (r = -0.510). However, there were no significant changes in plasma zinc and copper values during OGTT in both obese individuals and lean controls. This study showed that plasma zinc values had no changes during OGTT in obese individuals. The results also indicated that lower fasting plasma zinc concentrations in obese individuals were not the short-term metabolic result. PMID- 9404683 TI - Depressed antioxidant defense in rat heart in experimental magnesium deficiency. Implications for the pathogenesis of myocardial lesions. AB - Magnesium (Mg) deficiency has been shown to produce myocardial lesions in different experimental models. Based on several lines of evidence, it has been proposed that oxidative injury to the cardiac muscle may explain the pathobiology of such lesions. In pursuance of this postulation, the present study examined the effect of dietary deficiency of Mg on the activity of the antioxidant enzymes, superoxide dismutase (SOD) and catalase, in rat heart. This article reports a significant lowering of the activity of both these enzymes in the cardiac tissue in Mg-deficient rats. Since depressed antioxidant defense in the heart may enhance myocardial susceptibility to oxidative injury, the observation is of possible relevance to the pathogenesis of cardiac lesions in Mg deficiency. PMID- 9404682 TI - Assisting effects of lithium on hypoglycemic treatment in patients with diabetes. AB - In this article, we report the assisting effect of lithium on hypoglycemic treatment in patients with diabetes. Thirty-eight diabetic patients, 15 male and 23 female, aged 20-70 yr, 33 noninsulin-dependent diabetes mellitus (NIDDM) patients, and 5 insulin-dependent diabetes mellitus (IDDM) patients, were recruited in this study. Fasting and 1-h postprandial blood glucose (BG) profiles were undertaken from three groups of patients with diabetes before and after short-term of treatment of lithium carbonate. Group I was treated with diet only, Group II with oral hypoglycemic agents (OHA), and Group III with insulin. The fasting blood glucose (FBG) level and 1-h postprandial blood glucose (1-h PBG) level before and after treatment of lithium were: Group I: FBG: 7.67 +/- 0.48 vs 7.13 +/- 0.82; 1-h PBG 15.13 +/- 0.88 vs 10.33 +/- 0.96; Group II: FBG: 8.84 +/- 0.67 vs 6.04 +/- 0.57; 1-h PBG: 12.33 +/- 0.72 vs 9.95 +/- 0.82; Group III: FBG: 10.87 +/- 0.83 vs 6.83 +/- 0.79; 1-h PBG: 12.45 +/- 0.93 vs 9.17 +/- 1.00 mmol/L, respectively. The FBG and PBG of all three groups decreased significantly after lithium treatment, except the FBG in Group I. These data suggest that combined with other therapy, lithium could improve glucose metabolism in most patients with diabetes. Our results suggest that lithium has an assisting hypoglycemic effect on antidiabetic treatment. PMID- 9404685 TI - Multivessel system for cold-vapor mercury generation. Determination of mercury in hair and fish. AB - A multivessel system for the determination of mercury (Hg) by cold-vapor atomic absorption spectrometry (CV-AAS) and inductively coupled plasma atomic emission spectrometry (ICP-AES) was developed. The performance of the proposed device was tested by determining total Hg in quality-control samples of hair and fishes following acid digestion. Application of the apparatus to the determination of Hg by CV-AAS following alkaline digestion was studied as well. The detection limit obtained for CV-AAS was 0.11 ng/mL and for ICP-AES 1.39 ng/mL. The results show that the system is appropriate to be used in techniques involving cold-vapor generation of Hg. PMID- 9404684 TI - Trace metals and metalloenzymes in placenta after oral administration of lead acetate. AB - The present study was carried out to find the effects of Pb acetate (10-50 mg/kg body wt) after oral administration on: 1. The distribution of elements, such as Fe, Cu, Zn, and Mn; 2. The activity of 6-amino levulenic acid dehydratase (delta ALAD) and alkaline phosphatase (PAP); and 3. On the level of reduced glutathione (GSH) in murine placenta. Pb toxicity expressed on a dry-wt basis was reflected in terms of deficiency of delta-ALAD and PAP and enhanced content of GSH. Analysis of trace elements following Pb exposure showed low levels of Mn and Cu. Although Fe composition of placenta remained within normal range with increasing load of endogeneous Pb, Zn decline was not consistent after oral feeding of Pb acetate. Deficiency of PAP after Pb exposure did not correlate with the endogeneous levels of Pb or Zn therein, but correlated with endogeneous levels of Mn. Placental deficiencies of Cu and Mn have been related to the disturbed placental functions by Pb accumulation. PMID- 9404686 TI - Historical aspects and definition of vitiligo. PMID- 9404687 TI - Pathophysiology of vitiligo. PMID- 9404688 TI - Immune mechanisms in vitiligo. PMID- 9404689 TI - Microscopic changes in vitiligo. PMID- 9404690 TI - The epidemiology and genetics of vitiligo. PMID- 9404691 TI - Vitiligo: a psychologically influenced and influencing disease. PMID- 9404693 TI - Childhood vitiligo. PMID- 9404692 TI - Clinical features of vitiligo. PMID- 9404694 TI - Non PUVA nonsurgical therapies for vitiligo. PMID- 9404695 TI - Psoralen photochemotherapy for vitiligo. PMID- 9404696 TI - Surgical therapies for vitiligo. PMID- 9404697 TI - Angiogenesis in ovarian carcinoma: a formidable biomarker. PMID- 9404698 TI - A randomized, controlled phase III study of cyclophosphamide, doxorubicin, and vincristine with etoposide (CAV-E) or teniposide (CAV-T), followed by recombinant interferon-alpha maintenance therapy or observation, in small cell lung carcinoma patients with complete responses. AB - BACKGROUND: Studies of chemotherapy for patients with small cell lung carcinoma (SCLC) have shown that teniposide (T) may have higher activity than etoposide (E). In this randomized, controlled Phase III study, the authors compared cyclophosphamide, doxorubicin, and vincristine (CAV) with E and CAV with T as induction treatments for patients with SCLC. A second objective of the study was to study patients who had achieved complete response (CR). They were considered for a second randomization to maintenance therapy, in which they would receive either recombinant interferon-alpha (rIFN-alpha) or no treatment. METHODS: From June 1990 to December 1995, 140 untreated SCLC patients were enrolled in this study. Patients were stratified by either limited disease (LD) or extensive disease (ED) and randomized to one of two treatment arms. The schedules for both arms included cyclophosphamide 1000 mg/m2 administered intravenously (i.v.), doxorubicin 50 mg/m2 i.v., and vincristine 2 mg i.v. on Day 1. Arm A (CAV-E) involved the addition of E 100 mg/m2 i.v. on Days 2, 3, and 4; Arm B (CAV-T) involved the addition of T 60 mg/m2 i.v. on Days 2, 3, and 4. Courses were repeated every 3 weeks. After 3 courses, patients with LD received chest radiotherapy and 2 additional consolidation courses, whereas patients with ED received 5 consecutive courses only. Patients with CR were considered for the second randomization, which consisted of either maintenance therapy with intramuscular (i.m.) rIFN-alpha-2b, 3 M.U., once a day for 9 months (IFN-alpha arm) or no therapy (control arm). RESULTS: At 5 years from start-up (3-year median observation time and 90% death rate), the study was closed. Results were as follows: 140 patients (71 in Arm A and 69 in Arm B) were eligible for survival analysis; 131 were evaluable for response and toxicity (66 in Arm A and 65 in Arm B), whereas 9 were not (6 early deaths and 3 with protocol violations). Among evaluable patients, 68 showed LD (35 assigned to Arm A and 33 to Arm B); the responses to treatment were 28.5% (10/35) CR and 51% (18/35) partial response (PR) to CAV-E, and 39% (13/33) CR and 39% PR (13/33) to CAV-T. Sixty-three patients showed ED (31 assigned to Arm A and 32 to Arm B); their responses were 22.5% (7/31) CR and 52% (16/31) PR to CAV-E, and 12.5% (4/32) CR and 50% (16/32) PR to CAV-T. Drug-related toxicity was WHO Grade 3-4 myelosuppression in 20% of 292 CAV-E courses and in 27% of 252 CAV-T courses. There were 6 toxic deaths, 1 in Arm A and 5 in Arm B (chi-square = 2.86); 2 patients in Arm A discontinued therapy due to persistent leukopenia and thrombocytopenia. No other remarkable toxicities were observed. Actuarial median survival (MS) was 13.7 months (range, 1.0-62.5 months) for patients with LD receiving CAV-E (Arm A) and 15.2 months (range, 0.5-68.2 months) for those receiving CAV-T (Arm B) (chi-square = 0.89); in patients with ED it was 10.5 months (range, 0.6-30.4 months) and 8.2 months (range, 0.2-24.8 months), respectively (chi-square = 3.42). Overall, MS was 12 months (range, 0.6-62.5 months) in Arm A and 10 months (range, 0.2-68.2 months) in Arm B (chi-square = 0.059). Thirty-nine patients with CR (27.8%) were candidates for the second randomization. Among them, 26 patients (18.5%) complied with the program and were randomized as follows: 14 were assigned to the IFN alpha arm and 12 to the control arm. Starting from the second randomization, median time to progression was 12 months (range, 3-51 months) for patients in the IFN-alpha arm versus 7 months (range, 1-59 months) for patients in the control arm (chi-square = 0.12). MS was 15 months (range, 5-52.3 months) versus 9 months (range, 2-60.5 months) (chi-square = 0.13). CONCLUSIONS: This study did not show a wide difference in activity and toxicity between CAV-E and CAV-T. The number of patients who entered the second randomization was too small to reach the second study endpoint. PMID- 9404699 TI - Expression of transforming growth factor-beta (TGF-beta) isoforms in osteosarcomas: TGF-beta3 is related to disease progression. AB - BACKGROUND: Transforming growth factor-beta (TGF-beta) is a multipotent growth factor affecting development, homeostasis, and tissue repair. In addition, increased expression of TGF-beta has been reported in different malignancies, suggesting a role for this growth factor in tumorigenesis. METHODS: Using immunohistochemistry, the expression, prevalence, and distribution of TGF-beta isoforms were evaluated in 25 high grade human osteosarcomas. The Cox proportional hazards models and Kaplan-Meier curves were calculated correlating disease free survival with TGF-beta expression. RESULTS: Expression of one or more TGF-beta isoforms was found in all the osteosarcomas. Immunoreactivity for TGF-beta1 and TGF-beta3 generally was stronger than for TGF-beta2. The cytoplasm of the tumor cells showed stronger staining than their surrounding extracellular stroma. Most notably, osteoclasts showed strong to intense staining for all three isoforms. In 11 of 25 specimens angiogenic activity was noted with staining of multiple small vessels in the tumor stroma. Expression of TGF-beta3, but not of TGF-beta2 or TGF-beta1, related to disease progression, such that there was a statistically significant decrease in the disease free interval as the immunoreactivity for TGF-beta3 increased. CONCLUSIONS: All osteosarcomas expressed TGF-beta in the cytoplasm of the tumor cells as well as in their extracellular stroma. The presence of TGF-beta in the endothelial and perivascular layers of small vessels in the tumor stroma suggests angiogenic activity of this growth factor. The expression of TGF-beta3 was correlated strongly with disease progression (P = 0.027). These data suggest that increased expression of TGF-beta isoforms, especially TGF-beta3, may play a role in osteosarcoma progression. PMID- 9404700 TI - The prognostic significance of sialyl-Tn antigen in women treated with breast carcinoma treated with adjuvant chemotherapy. AB - BACKGROUND: Sialyl-Tn (STn) represents an aberrantly glycosylated mucin epitope that is expressed in breast carcinoma and other adenocarcinomas and is an important factor in the development of novel immunotherapeutic approaches. The primary aim of the current study was to investigate the influence of STn expression on the prognoses of patients with breast carcinoma. METHODS: A cohort of 207 women diagnosed with invasive breast carcinoma who were treated with anthracycline-containing adjuvant chemotherapy and were enrolled in a randomized clinical trial were studied. Expression of STn was determined by an immunohistochemical procedure in which the B72.3 monoclonal antibody was used. Kaplan-Meier and Cox proportional regression survival analyses were used to compare low STn and high STn patients. RESULTS: Forty-eight (23%) of the 207 specimens demonstrated high STn staining (>25% cells were immunoreactive). During a median follow-up of 5 years, high STn patients had worse disease free survival than low STn patients (55% vs. 74%, respectively; P = 0.03). High STn expression was significantly associated with age (P = 0.04) but not with other conventional prognostic markers. In multivariate analysis using the Cox regression model, high STn emerged as an independent prognostic indicator for disease free survival (hazard ratio [HR], 2.02; 95% confidence interval [CI], 1.09-3.73) and for overall survival (HR, 2.16; 95% CI, 0.95-4.92). CONCLUSIONS: The results of this study suggest that STn may be a valuable marker for identifying women at high risk of developing recurrent breast carcinoma who may be candidates for trials investigating new therapies in combination with standard adjuvant therapy. PMID- 9404701 TI - Immunohistochemical analysis of the expression of cdk4 and p16INK4 in human endometrioid-type endometrial carcinoma. AB - BACKGROUND: Abnormalities in G1 cell cycle regulation have been associated with the malignant transformation of cells. To obtain further information about the role of factors regulating the G1 cell cycle in the development of endometrial carcinoma, the authors analyzed the expression of cdk4 (cyclin-dependent kinase) and p16INK4 (an inhibitor of cdk4). METHODS: Immunohistochemical staining was performed on 20 specimens of normal endometria and 41 specimens of endometrioid type endometrial carcinoma using antibodies against cdk4 and p16INK4. RESULTS: In the glandular epithelia of the normal endometria, cytoplasmic staining of cdk4 and p16INK4 was observed only in the proliferative phase, but nuclear staining of these agents was negligible. In endometrial carcinomas, 8 (19.5%) and 14 (34.2%) were positive for cdk4 and p16INK4 in the nucleus, respectively. Topographically, the nuclear cdk4 positive tumor cells were negative for p16INK4 and the nuclear p16INK4 positive tumor cells were found in areas without nuclear cdk4 expression, suggesting an inverse correlation between the two agents. In addition, the poorly differentiated carcinomas were more frequently positive for nuclear cdk4 than were the highly differentiated carcinomas (P = 0.03). CONCLUSIONS: These data suggest that increased expression of nuclear cdk4 associated with loss of p16INK4 expression could be involved in the carcinogenesis of a subset of endometrial carcinomas. PMID- 9404702 TI - Tumor angiogenesis as a prognostic factor in ovarian carcinoma: quantification of endothelial immunoreactivity by image analysis. AB - BACKGROUND: The growth of a malignant tumor requires the formation of new capillaries. Quantification of these microvessels is difficult. The purpose of this study was to establish an objective technique for quantifying angiogenesis and to evaluate whether microvessel quantity may predict tumor aggressiveness in patients with ovarian carcinoma. METHODS: Endothelial area was used to quantify microvessel density in immunohistochemically stained sections of 28 International Federation of Gynecology and Obstetrics Stage IIIC ovarian carcinomas. The endothelial area was measured with a computer-aided image analysis system in the subepithelial stroma of highest vascularization. The endothelial area in the specimens of 14 patients who survived for > or =6 years was compared with that of 14 patients matched for stage and treatment who died of the disease. RESULTS: The mean tumor area analyzed was 5.04 +/- 0.23 mm2. The mean endothelial area per mm2 of stroma from survivors and dead patients was 0.038 +/- 0.026 mm2 and 0.110 +/- 0.034 mm2, respectively (P < 0.0001). No significant differences were found in histology, tumor grade, status of lymph nodes, and amount of residual tumor. CONCLUSIONS: Image analysis was used to overcome the potential subjectivity of manual counts. Computer-assisted image analysis can evaluate accurately the angiogenic potential in ovarian carcinomas. Tumor angiogenesis may prove to be a prognostic factor in patients with ovarian carcinoma. This study suggests that the measurement of the endothelial area would be clinically useful in determining microvessel density [See editorial on pages 2219-21, this issue.] PMID- 9404703 TI - The cytotoxic effect of fleroxacin and ciprofloxacin on transitional cell carcinoma in vitro. AB - BACKGROUND: There have been few reports concerning the cytotoxic effects of fluoroquinolone antibiotics on transitional cell carcinoma. This investigation was designed to study the cytotoxic effects of fleroxacin and ciprofloxacin on transitional cell carcinoma quantitatively in vitro. METHODS: Two transitional cell carcinoma cell lines, MBT-2 and T24, were used in this study. The effects of fleroxacin and ciprofloxacin on cell proliferation were determined by counting the number of living cells and by colorimetric MTT assay. RESULTS: Two fluoroquinolones, fleroxacin and ciprofloxacin, significantly inhibited cell proliferation in a dose-dependent manner at a concentration of 50-800 microg/mL in both cell lines. Compared with the cytotoxic effects of the two antibiotics, the inhibitory activity of ciprofloxacin on cell proliferation significantly exceeded that of fleroxacin in the MBT-2 cell line. However, the two fluoroquinolones did not have significantly different effects on the T24 cell line. CONCLUSIONS: Fleroxacin and ciprofloxacin significantly affect cell proliferation in transitional cell carcinoma cell lines. The results encourage further study of the possibility of clinical application of some fluoroquinolones to prevent recurrence of urinary bladder tumors because the urinary excretions after oral administration of these drugs are quite high. PMID- 9404704 TI - Extracapsular invasion of lymph node metastasis is an indicator of distant metastasis and poor prognosis in patients with thyroid papillary carcinoma. AB - BACKGROUND: In patients with thyroid papillary carcinoma, age and the presence or absence of distant metastasis are regarded as the main prognostic factors. However, the histologic characteristics of thyroid papillary carcinoma that develops distant metastasis have not yet been clarified. METHODS: The histologic findings and prognosis of 50 patients with thyroid papillary carcinoma who later developed distant metastasis (metastatic group) were compared with those of 50 patients without local recurrence or distant metastasis (control group). The age, tumor size, and gender ratio of the control group were matched with those of the metastatic group. Univariate analyses (chi-square test and/or Fisher's exact test) and multivariate analyses (logistic regression) were performed. RESULTS: Univariate analyses showed that the incidence of nonpure papillary carcinoma, absence of bone at the periphery of the tumor, invasion of the perithyroidal muscle, large lymph node deposits, and extranodal invasion were significantly higher in the metastatic group. Multivariate analyses revealed that only extranodal invasion was statistically significant (P = 0.0045) and that the odds ratio of extranodal invasion in distant metastasis was 9. Moreover, the risk of death from thyroid carcinoma was higher among the patients with extranodal invasion than those without (P <0.01). CONCLUSIONS: The presence of extranodal invasion in patients with thyroid papillary carcinoma is an indicator of distant metastasis and poorer prognosis. PMID- 9404705 TI - Extended field and total central lymphatic radiotherapy in the treatment of early stage lymph node centroblastic-centrocytic lymphomas: results of a prospective multicenter study. Study Group NHL-fruhe Stadien. AB - BACKGROUND: A prospective multicenter trial was performed to evaluate survival, patterns of relapse, and toxicity for clinically staged patients with lymph node centroblastic-centrocytic (cb/cc) lymphomas in Stages I-IIIA after large extended field irradiation (EFI) or total central lymphatic irradiation (TCLI). METHODS: Between January 1986 and August 1993, 117 adults with clinical Stage I-IIIA lymph node cb/cc lymphoma (Kiel classification) were recruited. Patients in Stages I or II with mediastinal, hilar, periaortic, iliac, or mesenteric involvement and in Stage IIIA received TCLI, whereas patients with more peripherally located cb/cc lymphomas were treated with EFI. TCLI and EFI were administered to a total dose of 26 gray (Gy) with 2 Gy per daily fraction, with the exception of the whole abdomen, which was irradiated to a total dose of 25.5 Gy with 1.5 Gy per fraction. A boost of 10 Gy with 2 Gy per fraction was administered to enlarged and involved lymph nodes at the start of radiotherapy. RESULTS: Sixty, 40, and 17 patients had Stage I, II, and limited IIIA disease (no bulk and less than 6 involved lymph node regions), respectively. Overall survival was 86% at 5 and 7 years; median follow-up was 68 months. The probabilities of relapse at any site, recurrences in lymph nodes, and in-field lymph node recurrences after TCLI were 17% in Stage I; 56%, 43%, and 40% in Stage II, respectively; and 44%, 35%, and 35% in Stage IIIA, respectively. The risk of disseminated extralymphatic relapses was 9% at 7 years. The most important adverse prognostic factor for in-field lymph node recurrences was a deviation of >20% from the assigned total radiation dose. After EFI, patients in Stage I had a significantly lower risk of recurrences in adjuvant irradiated lymph node regions than in unirradiated lymph node regions. Acute toxicity of EFI and TCLI was moderate. CONCLUSIONS: In-field lymph node recurrences remained the main risk after TCLI, and a deviation of >20% from the assigned radiation dose was the major risk factor for in-field recurrences. From these data, a total dose of 40-44 Gy in conventional fractionation for the treatment of macroscopic cb/cc lymphomas and 30 Gy for the treatment of subclinical disease is recommended. A randomized study comparing TCLI with EFI is now being organized by this group. PMID- 9404706 TI - Intensified therapy for infants with acute lymphoblastic leukemia: results from the Dana-Farber Cancer Institute Consortium. AB - BACKGROUND: Infants with acute lymphoblastic leukemia (ALL) have a very poor prognosis. Since 1985, we have intensified therapy for infants with ALL by including a month of high dose multiagent chemotherapy after remission induction. METHODS: Between 1985 and 1995, we treated 23 infants (age < 12 months). We compared the presenting characteristics and outcomes of these infants with the 11 infants treated on our protocols between 1973 and 1985, an era prior to the intensification of therapy. Available bone marrow samples from infants treated since 1985 were analyzed for the presence of MLL gene rearrangements by Southern blot analyses and for TEL-AML1 gene fusion by reverse transcriptase-polymerase chain reaction. RESULTS: With a median follow-up of 5.6 years, the 50-month event free survival (EFS) (+/- standard error) for the 23 infants was 54 +/- 11%, a significant improvement (P = 0.001) compared with the outcome for the 11 infants treated on our protocols prior to 1985 (EFS = 9 +/- 9%). Of the seven infants found to have a rearranged MLL gene, three (43%) remained in first complete remission. None of the nine infant bone marrow specimens tested had evidence of TEL-AML1 gene fusion. The intensified therapy was complicated by a high incidence of infections, including septicemia in 52% of patients and Pneumocystis carinii pneumonitis in 22% of patients. Late effects identified in the 13 long term survivors (median age, 6 years) included developmental delay and learning disabilities of varying severity (82% of evaluable patients), asymptomatic cataracts (67%), asymptomatic echocardiographic abnormalities (30%), obesity (27%), and short stature (18%). CONCLUSIONS: Intensification of therapy significantly improved the EFS of infants with ALL compared with previous, less intensive regimens and with the experience of other investigators. Future treatment for infants should attempt to improve efficacy while minimizing toxicity. PMID- 9404708 TI - The National Cancer Data Base report on recent hospital cancer program progress toward complete American Joint Committee on Cancer/TNM staging. AB - BACKGROUND: American Joint Committee on Cancer (AJCC) staging procedures were first published in 1977. Since 1991 the Commission on Cancer (COC) has required AJCC staging of all nonpediatric cancers. The National Cancer Data Base (NCDB) encouraged recording of AJCC staging through analyses of selected aspects of staging completeness. We reviewed the trend toward the adoption of routine AJCC staging by hospitals for the 5-year period 1990-1994. METHODS: NCDB reports for nearly 2 million stageable cancers diagnosed from 1990 through 1994 were examined with emphasis on the hospital cancer program environment. Staging was complete if the hospital submitted stage codes for > or =90% of stageable cases or absent if stage codes were submitted for <5%. Hospitals were classified by ownership and type of cancer program. Regional staging practices also were reviewed. RESULTS: Overall staging increased from 78% to 88%, with increases for every site except carcinomas of the skin, cancers of the extrahepatic bile ducts and urethra, melanoma of the eyelid, and retinoblastoma The percent of hospitals staging completely increased from 49% to 61%, and the percent not routinely staging decreased from 6% to 3%. Complete staging increased in all hospital categories except For-Profit. The trend toward complete staging was uneven among states and regions. CONCLUSIONS: Hospital staging policies were affected by activities of the AJCC, COC, NCDB, clinical protocol procedures, and state policies. The varied completeness of staging at the hospital level by state, region, and type of hospital indicates that the adoption of routine staging is ongoing. PMID- 9404707 TI - The growth and maturation of the National Cancer Data Base. AB - BACKGROUND: The National Cancer Data Base (NCDB), a joint project of the Commission on Cancer of the American College of Surgeons and the American Cancer Society, is a cancer management and outcomes data base for health care organizations. It provides a comparative summary of patient care that is used by communities and participating hospitals for self-assessment. The most current (1994) data are described here. METHODS: Six calls for data have yielded a total of 4,580,000 cases for the years 1985-1994. A total of 1735 hospital cancer registries have each participated in at least one of the calls for data. RESULTS: Summing the last year's report from each of the 1227 hospitals that participated in 1994, the cases represent the equivalent of 57% of the estimated 1994 U.S. cancer cases. These data were received from all six regions of the country, including all 50 states. Ninety-seven percent of patients received all or part of their treatment at the reporting hospital. The four most common cancers are carcinomas of the breast (15.7%), lung (14.3%), prostate (13.1%), and colon (7.7%), and collectively they comprise a majority of new cases. CONCLUSIONS: The NCDB is a cancer management and outcomes data base for health care organizations that currently provides data on 57% of the estimated new cases in the U.S. Past data have been used extensively to assess patterns of care and outcomes. PMID- 9404709 TI - The National Cancer Data Base report on non-Hodgkin's lymphoma. AB - BACKGROUND: The National Cancer Data Base (NCDB) has reported on many malignancies occurring in men and women in the U. S. from >1400 contributing hospitals. The current report on non-Hodgkin's lymphoma (NHL) is a companion to an upcoming Patient Care Evaluation study of this relatively common and serious cancer. METHODS: This report is comprised of all NHL cases submitted to the NCDB divided into two diagnostic-year groups: 1985-1988 and 1990-1993. Variables routinely collected by hospital cancer registries have been analyzed to report on patterns of diagnosis and treatment. RESULTS: High grade NHL cases were more likely to be Stage IV (40.8%) than were low or intermediate grade cases (34.8% and 32.5%, respectively). Patients with NHL arising from lymph node sites tended to present with more advanced disease (55.8% with Stages III and IV disease), whereas patients with NHL arising from extranodal sites and non-lymph node nodal sites presented at an earlier stage (64.7% and 74.0%, respectively, with Stage I or Stage II disease). Approximately 67% of all patients underwent chemotherapy, whereas only 25% underwent surgery or radiation. By histology, 5-year survival was 68.8% for low grade disease, 51.9% for intermediate grade disease, and 45.8% for high grade disease; by stage, survival rates ranged from 73.5% for Stage I to 42.9% for Stage IV disease. CONCLUSIONS: To the authors' knowledge, the 91,306 cases in this study represent the largest contemporary sample of NHL patients. The material reported here may serve as a reference with which to compare local patterns with national data. The Working Formulation's ability to stratify patients' survival rates confirms its utility for NHL. Stage according to the American Joint Committee on Cancer also was accurate in predicting survival. PMID- 9404710 TI - The National Cancer Data Base report on patterns of childhood cancers in the United States. AB - BACKGROUND: Patterns of and progress against childhood cancer have been reported on multi-institution, regional, national, and international bases by several sources in the past. These sources have included clinical cooperative group trials and population-based registries. In general, the population-based surveys have excluded brain tumors of either benign or uncertain behavior. The authors of this article investigated the patterns of data reported for the period 1985-1993, motivated by their interest in assessing the potential of National Cancer Data Base (NCDB) data to 1) facilitate individual institution review and 2) cover institutions that are not members of the Pediatric Oncology Group or the Children's Cancer Group, which are both national clinical cooperative groups. METHODS: Six annual calls for data, starting with a call for 1985 and 1988 cases, were issued to approximately 2100 hospitals with cancer programs (1340 programs approved by the Commission on Cancer of the American College of Surgeons and 760 other programs). The baseline data items of the NCDB included patient demography, tumor characteristics, initial treatment, and follow-up. The data for each patient were coded in the traditional manner by trained cancer registrars before being transmitted to the NCDB in standard format. RESULTS: In the most recent year for which data were reported, the NCDB included 42% of all estimated U.S. childhood cancers. The cases were reported by institutions that were members of the Pediatric Oncology Group and the Children's Cancer Group as well as nonmember institutions. The distribution of diagnostic groups reported to the NCDB was generally similar to that reported to SEER, except for lymphomas and brain cancer (the NCDB series included benign as well as malignant brain tumors). The distribution of diagnostic groups reported to the NCDB did not change over the 9 year reporting period (1985-1993). With regard to ethnicity, the most varied distribution of diagnostic groups was found among African American patients. For many types of cancer, the survival of those patients reported to the NCDB was similar to that of patients included in the SEER population-based series. These cancers included Wilms' tumor (NCDB 89% vs. SEER 88%), non-Hodgkin's lymphoma (NCDB 74% vs. SEER 70%), soft tissue sarcomas (NCDB rhabdomyosarcomas 70% and sarcomas 79% vs. SEER soft tissue sarcomas 71%), and neuroblastoma (NCDB 58% vs. SEER 57%). CONCLUSIONS: The authors concluded that the number of brain tumors of benign and uncertain behavior being diagnosed were significant enough in number that they should be included in regional and national cancer registries that report data for clinical purposes. They further concluded that for reasons of data inclusion and institutional coverage, the NCDB will be an important data base for pediatric cancers that will warrant increased use by pediatric investigators. PMID- 9404711 TI - The National Cancer Data Base report on gastric carcinoma. AB - BACKGROUND: The National Cancer Data Base (NCDB) represents a national electronic registry system now encompassing almost 60% of incident cancers in the United States. In combination with other programs of the American College of Surgeons Commission on Cancer, the NCDB offers a working example of voluntary, accurate, and cost-effective "outcomes management" on a both a local and a national scale. METHODS: For the accession years 1985-1993, the NCDB has obtained information on demographics, patterns of care, disease stage, treatment, and outcome for a convenience sample of 57,407 gastric carcinoma cases (1.6% of total NCDB cases). In addition to describing trends, this report focuses on 5-year relative survival for a cohort of 1987-1988 cases staged according to the third edition of the American Joint Committee on Cancer's TNM classification, as well as patterns of care for a cohort of 1992-1993 cases. RESULTS: Stage-stratified 5-year relative survival for the 1987-1988 cohort was as follows: IA, 71%; IB, 56%; II, 37%; IIIA, 18%; IIIB, 11%; IV, 5%. Without noteworthy changes in stage distribution, demographics, or other factors, the proportion of patients treated by total gastrectomy is increasing slightly, but proximal gastrectomy for proximal cancers remains surprisingly popular. The proportion of cases receiving postoperative adjuvant treatment has declined slightly. Presumably because of advanced age and/or medical infirmity, a substantial proportion of U.S. patients with disease at every stage receive no treatment for cancer. CONCLUSIONS: This analysis of patterns of care has revealed unexplained variations in treatment and opportunities for improvement. Treatment of the elderly, infirm patient with gastric carcinoma appears problematic. PMID- 9404712 TI - Cell and molecular neurobiology of presenilins: a role for the endoplasmic reticulum in the pathogenesis of Alzheimer's disease? AB - Mutations in genes encoding presenilin-1 (PS-1) and presenilin-2 (PS-2) cause many cases of autosomal dominant inherited forms of early-onset Alzheimer's disease (AD). PSs are expressed in neurons throughout the nervous system, with differences in abundance among cell populations. PS-1 and PS-2 each have six to eight transmembrane domains and are localized mainly in the endoplasmic reticulum (ER). PSs may interact with cytoskeletal proteins and beta-amyloid precursor protein (APP) in ways consistent with roles in membrane trafficking and APP processing. Expression of mutant PSs in cultured cells and transgenic mice results in increased production of an amyloidogenic-cytotoxic form of amyloid beta-peptide (Abeta). Neural cells expressing mutant PSs exhibit increased sensitivity to apoptosis induced by trophic factor withdrawal and Abeta. The proapoptotic action of mutant PSs involves perturbed calcium release from ER stores and increased levels of oxidative stress. PS mutations may also suppress neurotransmitter synthesis in cholinergic neurons, suggesting a role in regulation of neuronal phenotype. Homology of PSs with the C. elegans gene sel-12 and phenotypic similarities of PS-1 and Notch knockout mice suggest a developmental role for PSs in somitogenesis. Collectively, the emerging data suggest intriguing roles of PSs in neuronal plasticity and cell death and highlight the importance of the ER as a regulatory site involved in the pathogenesis of neuronal degeneration in AD. PMID- 9404713 TI - Activation of NMDA receptors and Ca2+/calmodulin-dependent protein kinase participate in phosphorylation of neurofilaments induced by protein kinase C. AB - Aberrant phosphorylation of neurofilaments, similar to that occurring in various motor neuron diseases, is produced in cultured motor neurons by activation of protein kinase C (PKC). Following exposure to synthetic diacylglycerol, persistent change in the phosphorylation state of C-terminal domains of neurofilament proteins was detected by increased perikaryal immunoreactivity with the antibody SMI34; this antibody recognizes NF-M/NF-H when C-terminal KSP repeat domains are highly phosphorylated. SMI34 labeling of perikarya and dendrites was prevented by pretreatment with either the NMDA receptor antagonist APV or by the Ca2+/calmodulin-dependent protein kinase (CaMK) inhibitor KN-62, but not by antagonists of AMPA/kainate or metabotropic glutamate receptors or by inhibitors of arachidonic acid metabolic pathways. Thus, activation of PKC may induce neurofilament phosphorylation in motor neurons by acting in cooperation with stimulation of NMDA receptors and activation of CaMK. These mechanisms may be relevant to motor neuron disease and other neuronal injuries in which increased PKC activity has been measured. PMID- 9404714 TI - 17Beta-estradiol attenuates oxidative impairment of synaptic Na+/K+-ATPase activity, glucose transport, and glutamate transport induced by amyloid beta peptide and iron. AB - Synapse loss, deposits of amyloid beta-peptide (Abeta), impaired energy metabolism, and cognitive deficits are defining features of Alzheimer's disease (AD). Estrogen replacement therapy reduces the risk of developing AD in postmenopausal women. Because synapses are likely sites for initiation of neurodegenerative cascades in AD, we tested the hypothesis that estrogens act directly on synapses to suppress oxidative impairment of membrane transport systems. Exposure of rat cortical synaptosomes to Abeta25-35 (Abeta) and FeSO4 induced membrane lipid peroxidation and impaired the function of the plasma membrane Na+/K+-ATPase, glutamate transporter, and glucose transporter. Pretreatment of synaptosomes with 17beta-estradiol or estriol largely prevented impairment of Na+/K+-ATPase activity, glutamate transport, and glucose transport; other steroids were relatively ineffective. 17Beta-estradiol suppressed membrane lipid peroxidation induced by Abeta and FeSO4, but did not prevent impairment of membrane transport systems by 4-hydroxynonenal (a toxic lipid peroxidation product), suggesting that an antioxidant property of 17beta-estradiol was responsible for its protective effects. By suppressing membrane lipid peroxidation in synaptic membranes, estrogens may prevent impairment of transport systems that maintain ion homeostasis and energy metabolism, and thereby forestall excitotoxic synaptic degeneration and neuronal loss in disorders such as AD and ischemic stroke. PMID- 9404715 TI - Similar pattern of MCP-1 expression in spinal cords and eyes of Lewis rats with experimental autoimmune encephalomyelitis associated anterior uveitis. AB - Monocyte chemoattractant protein-1 (MCP-1) is a member of the CC chemokine family responsible for the recruitment of T cells that have been found during inflammation of the spinal cord in experimental autoimmune encephalomyelitis (EAE) in Lewis rats immunized with myelin basic protein (MBP). Lewis rats injected with MBP also developed anterior uveitis (AU), which coincided with the onset of EAE. In the present studies, we examined the expression and distribution of MCP-1 in the eye and spinal cord during disease and compared it to the expression of Th1 cell type cytokines. Initially, MCP-1 expression was detected at the preclinical phase in the iris/ciliary body and lumbar spinal cord and increased during the course of EAE/AU. Mononuclear infiltrating cells and endothelial cells and astrocytes of the CNS could be identified as a source of MCP-1 by in situ hybridization. Kinetics of expression of Th1 characteristic cytokines such as IL-2 and IFNgamma was in agreement with the expression of MCP-1 chemokine. Moreover, induction of the gene expression of MCP-1 seemed to occur earlier than that of MIP-2, and it correlated with increasing disease severity. MCP-1 seems to contribute to the initial recruitment of inflammatory cells into both the tissues of the eye and CNS over the course of disease. PMID- 9404716 TI - Extracellular matrix proteins expressed by human adult astrocytes in vivo and in vitro: an astrocyte surface protein containing the CS1 domain contributes to binding of lymphoblasts. AB - Primary cultures of human astrocytes, expressing glial fibrillary acidic protein (GFAP), were obtained from postmortem brain tissue samples. These cultured astrocytes produced an extracellular matrix (ECM), containing laminin (Ln) and fibronectin (Fn), as shown with specific antibodies. The perinuclear staining observed in these cells indicated that these proteins were de novo synthesized. Monoclonal antibody (mAb) 90.45, which recognizes the CS1 sequence found in an alternatively spliced form of Fn, also stained cultured astrocytes. Immunohistochemical analysis of normal human brain tissue showed positive staining for the CS1 domain, both on protoplasmic and fibrous astrocytes located in the gray and white matter. In contrast to cultured astrocytes, no immunoreactivity for Ln or Fn was found on astrocytes in normal human brain tissue. These in situ data indicate that the CS1 domain expressed by astrocytes is not part of a splicing variant of Fn. Western blot analysis confirmed that the CS1 domain expressed by cultured human astrocytes is part of an astrocyte protein which is different from human Fn. The CS1 domain is a known ligand for the adhesion receptor alpha4beta1 (VLA-4). We found that the human lymphoma cell lines Jurkat and Ramos, which express alpha4beta1, bound to cultured human astrocytes, and that this interaction could be partly blocked by mAb 90.45 or a synthetic CS1 peptide. Thus, the novel CS1-containing surface protein expressed by astrocytes in vitro and in vivo, contributes to binding of lymphoblasts, and therefore may be a relevant adhesion molecule for the recruitment of alpha4 integrin expressing leukocytes into the central nervous system (CNS). PMID- 9404717 TI - Metabotropic glutamate receptors prevent nitric oxide-induced programmed cell death. AB - Activation of metabotropic glutamate receptor (mGluR) subtypes can prevent neuronal injury through the signal transduction pathways of nitric oxide (NO). It is this link to NO free radical injury and subsequent DNA damage that is the most intriguing. We therefore examined whether neuronal protection through mGluR activation was dependent on the molecular mechanisms of programmed cell death (PCD). The NO generators sodium nitroprusside and 3-morpholino-sydnonimine were administered to induce NO toxicity in primary hippocampal neurons. PCD was documented by hematoxylin and eosin nuclear staining, DNA gel electrophoresis, transmission electron microscopy, and protein synthesis assays. Following NO exposure, PCD induction was rapid and robust in approximately 70% of the neuronal population. Activation of specific mGluR subtypes with 1S,3R-ACPD and L-AP4, agents that are neuroprotective against NO, significantly limited the progression of PCD. In contrast, antagonism of mGluRs with L-AP3 did not prevent the development of PCD. Induction of new protein synthesis, a common requisite for PCD, was evident following NO exposure, but did not appear to represent a principal pathway of modulation by the mGluR agonists. Our studies suggest that mGluR modulation of NO-induced PCD represents a primary molecular pathway responsible for neuronal survival. Further elucidation of the molecular mGluR signaling pathways may yield new insight into specific genetic regulatory mechanisms responsible for neuronal injury. PMID- 9404718 TI - Arginine vasopressin (AVP) depletion in neurons of the suprachiasmatic nuclei affects the AVP content of the paraventricular neurons and stimulates adrenocorticotrophic hormone release. AB - Arginine vasopressin (AVP) produced in the hypothalamic suprachiasmatic nuclei (SCN) plays a role in establishing neuroendocrine rhythms and, in particular, in regulating the corticotrope axis rhythm. It has recently been shown that AVP from SCN inhibits corticosteroid release. In order to investigate the influence of suprachiasmatic AVP on the different peptidergic systems through the hypothalamus, SCN neurons containing AVP were functionally lesioned by using toxins associated with a cytotoxic monoclonal antibody (MAb) raised against AVP. Six days later, the AVP contents and AVP mRNA were measured in different hypothalamic and extrahypothalamic sites. Adrenocorticotrophic hormone (ACTH) concentration was also measured in plasma. Microinjection of the AVP-MAb/toxin mixture into SCN brought about a significant decrease in the AVP expression in SCN. This is demonstrated by the decrease in the AVP immunoreactive content (24%, P < 0.01) and the decrease of AVP hybridized mRNA (33%, P < 0.01). This points to the efficiency of the microinjection in decreasing the production of AVP in the injection area. Modifications of the AVP contents in the two subdivisions of the hypothalamic paraventricular nucleus (PVN) were also observed. AVP contents decreased in the parvocellular subdivision (pPVN); this is coherent with the AVP depletion in SCN since pPVN is the major site of the SCN hypothalamic efferences. AVP content and AVP mRNA increased in the magnocellular subdivision (mPVN); this also confirms the difference in AVP synthesis regulation according to the PVN subdivisions. The microinjection did not modify AVP expression in supraoptic nuclei or oxytocin (OT) immunoreactive content in the main hypothalamic OT containing sites. Plasma ACTH values were double (P < 0.02) the values measured under non-specific IgG treatment 10 hr after lights on. This probably resulted from the stimulation of the hypothalamo-pituitary-adrenal system since corticotrophin-releasing hormone (CRH) mRNA increased simultaneously by 24% (P < 0.05) in the PVN and the immunoreactive CRH content of the median eminence significantly decreased (26%, P < 0.05). Overall, our data confirm that AVP produced in the SCN inhibits the CRH-adrenocorticotrope axis in normal conditions, probably because of SCN projections of AVP neurons on the PVN. PMID- 9404719 TI - Responsiveness to depolarization of hypothalamic neurons secreting somatostatin under stress and estrous cycle conditions: involvement of GABAergic and steroidal interactions. AB - We studied the sensitivity to a depolarizing stimulus of hypothalamic fragments dissected from cycling female donor rats exposed or not to 30-min stress at 4 degrees C. The neuronal response was estimated in terms of the ability of tissue to release somatostatin when stimulated with 40 mM K+. The data showed no differences in response to K+, regardless of the ovarian cycle of the female donors, whereas tissues dissected from ovariectomized or pregnant rats responded significantly to K+. However, when donors underwent previous cold stress, significant differences were noted at all stages of the cycle, except diestrus-1, compared with control rats. We tested whether GABA and/or neuroactive steroids could be involved in this phenomenon and observed no GABA inhibition of somatostatin release in vitro, but inhibition occurred in the presence of a neuroactive steroid, THDOC. The effect of GABA in vivo on somatostatin release was estrogen dependent because bicuculline modified the total amount of somatostatin secreted in estrus but not in diestrus II. Finally, in hypothalamic primary cultures, GABA inhibition of somatostatin release was only detected when steroids were present in the media throughout culture. Our results suggest that steroid-GABA-somatostatin interactions could explain the different responses of neurons to depolarization. PMID- 9404720 TI - Regulation of the Na+-dependent high affinity glutamate/aspartate transporter in cultured Bergmann glia by phorbol esters. AB - The effects of phorbol 12-tetradecanoyl-13-acetate (TPA) and dibutyryl cAMP on the glutamate transport present in chick Bergmann glial cell (BGC) cultures were examined. TPA produced a significant decrease in [3H]-D-aspartate uptake, while dibutyryl cAMP treatment elicited a slight reduction in the transport. This effect was dose and time dependent and sensitive to staurosporine, a Ca2+/diacylglycerol-dependent protein kinase C (PKC) inhibitor. Long-term exposure of the culture to TPA results in a dramatic fall of the transporter activity and a decrease in the amount of the transporter protein. These findings suggest that PKC is involved in transport modulation and possibly in the regulation of the transporter gene expression. PMID- 9404721 TI - OlP-1, a novel protein that distinguishes early oligodendrocyte precursors. AB - Oligodendrocyte development may be divided into three distinct stages: I) commitment of neuroectoderm cells to the oligodendrocyte lineage, II) migration of precursors into the surrounding parenchyma concomitant with increased proliferation, and III) cessation of migration and proliferation and initiation of myelination. Stage II of development has remained enigmatic because of the paucity of known molecules that distinguish these immature migratory cells. We describe a novel surface protein, termed OlP-1, which is restricted in expression to this developmental stage in the mouse. Cytofluorographic comparisons with known developmental markers showed OlP-1 to be expressed primarily by stage II precursors in vitro. Histologic analyses supported this conclusion by showing co localization of OlP-1 with stage II molecules in vivo. Two conclusions were drawn from these results. First, OlP-1 was a novel protein expressed by murine oligodendrocyte precursors at a point in development that suggested a role in migration or proliferation. Second, dispersal of OlP-1-positive cells throughout the developing brain did not correlate with the location of myelination which, observed days later, progressed in a caudal to rostral manner. These data supported the concept that the final steps of maturation and myelin gene expression may be dependent upon extrinsic factors located predominantly within white matter tracts. PMID- 9404722 TI - Opposite effects of astrocyte-derived soluble factor(s) on the functional expression of fetal peptidergic neurons in aggregate cultures: enhancement of neuropeptide Y and suppression of somatostatin. AB - Previous studies established that fetal rat and human neuropeptide Y (NPY) cortical neurons in aggregate cultures are differentially regulated. Whereas brain-derived neurotrophic factor (BDNF) or phorbol 12-myristate-13-acetate (PMA) induces NPY production in rat cultures, only PMA does so in human cultures. We addressed these questions: 1) Do soluble products of rat or human astrocytes (conditioned medium; rCM and hCM, respectively) enhance the functional expression of cultured NPY neurons and if so, do they enhance the expression of somatostatin (SRIF) neurons as well? 2) Is the NPY-enhancing activity (EA) in the CM species specific? rCM enhanced (approximately 2-fold) both basal and BDNF-stimulated production of NPY and coculture of rat aggregates and astrocytes did not prevent this NPY-EA. Likewise, the hCM enhanced (approximately 2.5-fold) basal and PMA stimulated production of NPY by human aggregates. Moreover, the hCM enhanced NPY production by rat aggregates and rCM enhanced NPY production by human aggregates. In addition, rCM and hCM each enhanced BDNF-, forskolin-, or PMA-stimulated NPY production by rat aggregates. Under each of the above conditions, the rCM/hCM suppressed (approximately 50%) production of SRIF by rat aggregates. In summary, secretory products of rat and human astrocytes exert opposite effects on the functional expression of NPY and SRIF neurons in culture: enhancement of NPY and suppression of SRIF. By the criteria evaluated in this study, these astrocyte derived activities do not exhibit species specificity. PMID- 9404724 TI - Responses of cortical noradrenergic and somatostinergic fibres and terminals to adjacent strokes and subsequent treatment with NGF and/or the ganglioside GM1. AB - The occurrence of sprouting by fibre systems in the neocortex following lesion is still a controversial issue. In previous studies, we showed a nerve growth factor (NGF)-induced sprouting and hypertrophy of presynaptic terminals in the cholinergic fibres of the rat neocortex following stroke-type lesions, effects that were potentiated by the monosialoganglioside GM1. The present study investigated whether exogenous NGF and/or GM1 treatment could also affect the noradrenergic and somatostinergic systems in the neocortex. Immediately following unilateral vascular decortication, adult rats received, via minipump, a 7-day infusion of vehicle, NGF (12 microg/day) and/or GM1 (1.5 mg/day) into the cerebroventricular space. Thirty days postlesion, the animals were perfused with histological fixatives, the brains were removed, and relevant sections were processed for dopamine beta-hydroxylase and somatostatin immunocytochemistry at the light and electron microscopic levels. A Quantimet 920 image analysis system was used for the quantification of fibre length and size of presynaptic boutons. The lesion caused a reduction in the dopamine beta-hydroxylase-immunoreactive fibre length, which was not attenuated by either NGF or GM1 treatment or both. The somatostatin-immunoreactive network, in contrast, was unaffected by the lesion, and there was no sprouting of somatostatin fibres following trophic factor therapy. We also found no significant differences in the size and number of synapses of both the dopamine beta-hydroxylase-immunoreactive and somatostatin immunoreactive boutons following lesion and drug treatments. These results indicate that NGF and/or GM1 therapies do not cause regrowth in the noradrenergic and somatostatinergic cortical fibre networks or their presynaptic elements following a cortical devascularizing lesion. PMID- 9404723 TI - Increased expression of monoamine oxidase-B results in enhanced neurite degeneration in methamphetamine-treated PC12 cells. AB - In vivo administration of methamphetamine (MA) produces selective damage to dopaminergic nerve terminals, which is hypothesized to be due to release of dopamine from synaptic vesicles within the terminals, allowing the generation of reactive oxygen species (ROS) via dopamine metabolism. Hydrogen peroxide formed during this reaction can interact with free iron to form hydroxyl radicals, which can oxidize proteins, nucleic acids, and membrane lipids, leading to terminal degeneration. Elevation of activity of the dopamine-metabolizing enzyme monoamine oxidase (MAO) in nerve growth factor-treated PC12 cells resulted in a substantial rise in products of dopamine metabolism following MA treatment, including 3,4 dihydroxyphenylacetic acid and hydroperoxides, as well as an increase in lipid peroxidation and a decrease in neurite number and length compared with control cells. These latter effects could be reversed by treatment with the MAO-B specific inhibitor, deprenyl. These data suggest that dopamine metabolism and subsequent ROS production may be key elements in MA-induced neurite degeneration in dopaminergic neurons. PMID- 9404725 TI - Ganglioside lateralization in the brain of female rats. AB - The ganglioside composition of the cerebral hemispheres of young and adult rats of either sex has been herein assessed for the first time. In females, the total ganglioside content at any age, the content of GM1, GD1a, and GD1b at 8 days, and the content of GM1, GD1b, GT1b, and GQ1b at 60 days were higher in the right than in the left hemisphere. In males, no difference was observed. Concerning the ceramide moiety, a difference was displayed by C18:1 long-chain base in GD1a, whose proportion was higher in the left than in the right hemisphere of females aged 8 days. The comparison between homolateral hemispheres of rats of different sex revealed several differences. On average, in 8-day-old animals, the content of gangliosides was higher in females than in males. At 60 days the amount of gangliosides was on average lower in females than in males, even if with some exception. The data obtained with the current investigation show the existence of a ganglioside lateralization in rat brain, exclusively in females, and almost entirely at charge of the oligosaccharide portion. Moreover, age-dependent changes of ganglioside pattern and content show a dependence on brain lateralization. PMID- 9404726 TI - The trkC receptor is transiently localized to Purkinje cell dendrites during outgrowth and maturation in the rat. AB - In vivo studies of granule cell gene expression during corticocerebellar development and in vitro studies of Purkinje cell neurite outgrowth suggest that neurotrophin-3 may influence growth of Purkinje cell dendrites. To determine whether neurotrophic substances affect the growth of specific neuronal processes (i.e. axons and dendrites) or nonspecifically cause process development by exerting a trophic influence upon neuronal physiology we performed an immunohistochemical examination of trkC protein expression during early postnatal development of the rat cerebellum. Our findings indicate that Purkinje cells begin to synthesize trkC protein coincident with the onset of dendritic outgrowth. Robust immunostaining was evident throughout the entire somatodendritic domain of Purkinje cells during dendritic development but became faint and restricted to the cell body subsequent to the completion of dendritogenesis. These results suggest that growth and maturation of the Purkinje cell dendritic arbor may be influenced by neurotrophin-3 activation of trkC receptors distributed within developing dendrites. PMID- 9404727 TI - Neuronal protection from apoptosis by pituitary adenylate cyclase-activating polypeptide. AB - Pituitary adenylate cyclase-activating polypeptide (PACAP) is known to have trophic effects on neurons. Apoptosis of PC12 cells was induced by depletion of serum and nerve growth factor (NGF) from culture medium. Not only high potassium induced Ca2+ channel activation but PACAP-38 at physiological concentrations (10[ 10] to 10[-8] M) protected PC12 cells from apoptosis. PACAP-38 increased Ca2+ uptake and intracellular Ca2+ concentrations in PC12 cells. The effects of PACAP 38 on cell survival and Ca2+ channels were eliminated by inhibitors for Ca2+ channels and protein kinase A, and mimicked by 8-bromo-cAMP. Mitogen-activated protein (MAP) kinase activity was stimulated by PACAP-38. These findings implicate that PACAP protects PC12 cells from apoptosis by activating Ca2+ channels via the cAMP-protein kinase A pathway to stimulate MAP kinase cascade. PMID- 9404728 TI - Gastrin receptor expression and function during rapid transformation of the enterochromaffin-like cells in an African rodent. AB - The enterochromaffin-like cell (ECL) cells of the stomach are principally regulated by gastrin via a gastrin/CCK(B) receptor (G[R]) which modulates both histamine secretion and cell proliferation. In the African rodent (mastomys) hypergastrinemia generated by the histamine-2 receptor antagonist (loxtidine) results in ECL cell hyperplasia and neoplasia at 8 and 16 weeks respectively. The expression, structure and function of the G(R) during transformation is however unknown. We utilized a pure (approximately 90%) preparation of ECL cells to evaluate alterations in the G(R) utilizing immunocytochemistry, Western blot analysis, reverse transcription polymerase chain reaction (RT-PCR), 5-bromo-2 deoxyuridine uptake and phosphorylation site analysis. Although the expression of ECL cell G(R) was upregulated at both mRNA (PT-PCR) and protein (Western analysis) level, its affinity to gastrin was decreased in the hyperplastic phase and lost during transformation. The coding sequence of the G(R) of mastomys tumor ECL cells was identical to that of normal ECL cells, parietal cells and the brain. However, the mRNA sequence of the third introcytoplasmic loop of the G(R) was significantly different to other species. In addition, the G(R) exhibited phosphorylation site on serine residue(s). We have thus noted a direct correlation between hypergastrinemia and G(R) alteration and function during ECL cell transformation. It is possible that the unique mastomys gastrin receptor mediated ECL cell transformation involves the novel phosphorylation sites and a divergence in the introcytoplasmic domain. PMID- 9404729 TI - Human neuroblastoma cells use either insulin-like growth factor-I or insulin-like growth factor-II in an autocrine pathway via the IGF-I receptor: variability of IGF, IGF binding protein (IGFBP) and IGF receptor gene expression and IGF and IGFBP secretion in human neuroblastoma cells in relation to cellular proliferation. AB - Neuroblastoma cells are thought to depend upon autocrine stimulation by IGF-II but not by IGF-I. We have studied the expression of IGF, IGFBP and IGF receptor mRNA in two human neuroblastoma cell lines, SK-N-MC and CHP, and asked whether or not the expression of the IGF system in these malignant cells determines their growth pattern. SK-N-MC cells grow with a cell doubling time of 36 hours in medium supplemented with 10% fetal calf serum whereas CHP cells only grow with a doubling time of 72 h. In addition, the SK-N-MC cell line has a plating efficiency ten times greater than the CHP cell line. RNase protection assays were performed using (32)P-labelled riboprobes and RNA that had been purified from SK N-MC and CHP cells respectively. A 520 bases human IGF-I, a 556 bases human IGF II, a 480 bases human IGF-I receptor and a 250 human IGF-II/mannose-6-phosphate (M6P) receptor probe were radiolabelled as were human IGFBP-1, -2, -3, -4, -5 and -6 probes. While both SKNMC and CHP neuroblastoma cells expressed mRNAs for IGFBP 2, -4, and -6 no signal was detected for IGFBP-1, and -3 and only SK-N-MC cells expressed IGFBP-5 mRNA. In addition, a 400 bases protected band was seen with the IGF-I receptor probe and a 260 bases protected band with the IGF-IIM6P receptor probe in either cell line. Interestingly, a 300 bases protected species was detected with the IGF-II probe in CHP cell RNA whereas SK-N-MC cells did not express IGF-II transcripts. Conversely, SK-N-MC cells expressed a 520 bases IGF-I transcript while CHP cells did not show IGF-I mRNA expression. As determined by specific radioimmunoassays SK-N-MC cells secreted 0.75+/-0.02 ng/ml IGF-I, 1.2+/ 0.04 ng/ml IGF-II and 149+/-2.1 ng/ml IGFBP-2 within 24 h, whereas CHP cells secreted 0.1+/-0.01 ng/ml IGF-I, but 6.2+/-0.1ng/ml IGF-II and 254.8+/-5.5 ng/ml IGFBP-2 (N=5). IGFBP-2 secretion correlated positively with IGF-II secretion in CHP cells (r=0.85, P=0.05) and negatively with IGF-I (r= -0.9, P<0.01) in SK-N-MC cells. In conclusion, SK-N-MC cells which grow rapidly and have a high plating efficiency, express IGF-I, while CHP cells that grow more slowly express IGF-II. We hypothesize that neuroblastoma cells depend upon autocrine stimulation by either IGF-I or IGF-II. Variable sensitivity to growth inhibitors or apoptotic processes may be related to the differential expression of the IGF system. PMID- 9404730 TI - The potency of dietary amino acids in elevating plasma cholecystokinin immunoreactivity in cats is related to amino acid hydrophobicity. AB - Incomplete agreement exists on the relative potency of amino acids in stimulating endocrine secretion of cholecystokinin (CCK). Species and methodological variations have been suggested to account for the apparent inconsistencies. In the present research, the CCK-releasing potency of dietary amino acids was evaluated in cats using plasma CCK-like immunoreactivity (CCK-LI) as an indicator of CCK secretion rather than pancreatic protein and enzyme secretion, as has been used in past research. Oral-gastric administrations of a casein-simulating amino acid mixture increased (P < 0.05) plasma CCK-LI but not to the extent of that observed for casein or sodium oleate. The response in plasma CCK-LI to administrations of 50 mM solutions of amino acids was significant (P < 0.05) for tryptophan, phenylalanine, leucine, and isoleucine and the response increased linearly (P < 0.01) with increasing amino side-chain hydrophobicity. Control administrations of water and saline also evoked elevation in plasma CCK-LI, but the responses were so transient that amino acid effects were not obscured. This was substantiated by the finding of a significant linear (P < 0.001) dose response to phenylalanine administration. Cholecystokinin-8, 33 and 58 were among the CCK molecular forms identified by HPLC in plasma after administrations of phenylalanine and water. The present findings indicate that lipophilic amino acids released during digestion account for at least part of the endocrine CCK response in cats to ingested protein. The greater CCK-releasing potency observed for intact protein relative to free amino acids may have been the result of a slow digestive release of amino acids, elaboration of peptide secretogogues or protection of protease-sensitive releasing factors. PMID- 9404731 TI - Effect of vasoactive intestinal peptide (VIP) on cytokine production and expression of VIP receptors in thymocyte subsets. AB - Intrathymic T cell precursors undergo a programmed sequence of developmental changes resulting in the production of mature, self-MHC restricted, single positive T lymphocytes which migrate to the periphery. The intrathymic T cell development is controlled by various factors, including cytokines and possibly neuroendocrine hormones. Our previous studies indicate that vasoactive intestinal peptide (VIP) inhibits IL-2 and IL-4 production in thymocytes through different molecular mechanisms. Thymocytes acquire the competence to express IL-2 and IL-2R during thymic development in a maturation-dependent manner. In this study we investigate the effect of VIP on IL-2 production, and the expression of VIP-R1 and VIP-R2 mRNA in different thymocyte subsets in comparison to T cell lines. All thymocyte subsets and T cell lines tested express VIP-R2. In contrast, only single positive, CD4+8- and CD4-8+ thymocytes express VIP-R1. VIP inhibits IL-2 production in CD4+8+ and single positive CD4+8- and CD4-8+ thymocytes and in TH1 cells stimulated through the TCR. No inhibition is observed in CD3-4-8- and single positive CD4+8- and CD4-8+ thymocytes, or in TH1 cells stimulated by a combination of calcium ionophores and phorbol esters. These findings suggest that VIP inhibits IL-2 production through VIP-R2, and that it interferes with a TCR connected transduction pathway. We also investigate the expression of VIP mRNA in thymocyte subsets and T cell lines, and conclude that thymocytes as well as antigen-specific T cells may function as VIP sources within the lymphoid organs. PMID- 9404732 TI - Cyclic AMP modulates part of the relaxant action of calcitonin gene-related peptide in guinea pig gallbladder strips. AB - Calcitonin gene-related peptide (CGRP) has been shown to relax cholecystokinin induced tension in guinea pig gallbladder strips. This relaxation is dependent on the concentration of CGRP, and is primarily due to the opening of ATP sensitive K+ channels; however, other mechanisms may also be involved. Studies using forskolin, 8-bromoadenosine 3', 5' cyclic monophosphate, dibutyryl cAMP, cholera toxin, and Rp-adenosine 3', 5'-cyclic monophosphothioate triethylamine, which measured changes in tension suggest that cAMP may be involved in mediating the actions of CGRP. Radioimmunoassay of strips precontracted with cholecystokinin octapeptide (CCK) and either treated with CGRP or its solvent demonstrated that cAMP concentrations increased with CGRP treatment. The results of these studies demonstrate that CGRP acts through multiple mechanisms to induce relaxation of guinea pig gallbladder strips precontracted with CCK. PMID- 9404733 TI - Effect of islet amyloid polypeptide on somatostatin inhibition of insulin secretion from isolated rat pancreatic islets. AB - In this study, we investigated the presence of islet amyloid polypeptide (IAPP) in somatostatin cells of rat endocrine pancreas and the effect of exogenous IAPP and somatostatin, separate or combined, on in vitro insulin secretion. By immunocytochemistry, IAPP was found in both B and D cells of rat pancreatic islets. Furthermore, the labeling density of IAPP in D cells was nearly four times higher than in B cells. After a 2-day preincubation in RPMI 1640 (11.1 mM glucose), isolated rat pancreatic islets were exposed to IAPP and/or somatostatin for 90 min in modified Krebs-Ringer bicarbonate (KRB) buffers containing 11.1 or 22.2 mM glucose, or 11.1 mM glucose + 10 mM L-arginine, respectively. At 11.1 mM glucose, insulin secretion was not affected by IAPP and/or somatostatin at concentrations investigated. Insulin response to 22.2 mM glucose was inhibited by exogenous somatostatin. Arginine-stimulated insulin secretion was also inhibited by somatostatin, and the effect was significantly potentiated with additional 10( 5) M IAPP. The study shows that rat pancreatic D cells have higher IAPP density than B cells in the same islets and that IAPP and somatostatin may cooperate on rat pancreatic B cells to regulate the insulin secretion in response to potent stimulation. PMID- 9404734 TI - www.journal? Revisited. PMID- 9404735 TI - Flawed paradigms drive aerosol device selection. PMID- 9404736 TI - An analysis of platypnea-orthodeoxia syndrome including a "new" therapeutic approach. PMID- 9404737 TI - The antiphospholipid antibody syndrome: a vascular disease with pulmonary manifestations. PMID- 9404738 TI - Myocardial ischemia in sedated patients undergoing fiberoptic bronchoscopy. AB - STUDY OBJECTIVE: To study the incidence of myocardial ischemia and related hemodynamic alterations in sedated patients undergoing fiberoptic bronchoscopy (FOB). DESIGN: Prospective study. SETTING: Tertiary care, university hospital. PATIENTS: Twenty-nine patients, age 50 years or older, undergoing elective FOB. INTERVENTIONS: Myocardial ischemia was assessed by continuous ECG monitoring beginning 30 min before, and until 2 h after FOB. MEASUREMENTS AND RESULTS: During FOB, there was a significant rise in heart rate (89+/-3 [mean+/-SE] to 120+/-4 beats/min) and fall in oxygen saturation (95+/-1 to 90+/-1%). There was no significant rise in systolic or diastolic BP. Five patients (17%) had myocardial ischemia during FOB that lasted 20+/-8 min. Their demographic and pre FOB characteristics were not different from the other patients. Compared to baseline values, a significant rise in heart rate, a fall in oxygen saturation, and no significant change in BP were observed during FOB in patients, both with or without ischemia. Although not statistically significant, ischemia was associated with more protracted procedures. CONCLUSIONS: Myocardial ischemia may develop in elderly patients undergoing FOB. This observation encourages the routine use of ECG and oximetry during FOB, allowing for early intervention to prevent the dangerous combination of hypoxia, tachycardia, and myocardial ischemia. Moreover, this study suggests that methods to ensure oxygenation during FOB should be adhered to, and that the routine administration of atropine should be reconsidered. PMID- 9404739 TI - Diagnostic accuracy of transbronchial biopsy in acute farmer's lung disease. AB - STUDY OBJECTIVES: To evaluate whether transbronchial biopsy (TBB) is useful for the diagnosis of acute farmer's lung (FL) by calculating the likelihood ratios (LHRs) of (1) simple pathologic criteria and (2) an overall assessment of the biopsy specimens. DESIGN: Retrospective study in which a blinded analysis of 105 TBBs with adequate material from patients with parenchymal diseases (55 cases of FL matched with 50 control samples) was performed by two independent pathologists. SETTING: Respiratory clinic of a university-affiliated referral center. MEASUREMENTS: Three pathologic criteria were first studied: (1) diffuse lymphocytic infiltration (LI); (2) focal LI; and (3) granulomas. Then, an overall assessment or the TBB was done. Four diagnostic categories were considered: (1) probable FL; (2) possible FL; (3) nonspecific; and (4) alternative diagnosis. LHRs favoring the diagnosis of FL were calculated for the pathologic criteria and for each diagnostic category. RESULTS: For both the pathologic criteria and the overall assessments, the interobserver agreement was fair. As a pathologic criterion, "diffuse LI" was better than "loosely formed granuloma" to discriminate FL from control samples (LHR, 9.1 [confidence interval, 2.2 to 37.0] vs 1.8 [confidence interval, 0.5 to 6.9]). After the overall assessment, as many as 48.6% of the TBBs were read as nonspecific. The LHRs of the four diagnostic categories were as follows: (1) probable FL: 1.1 (observer 1) and 2.6 (observer 2); (2) possible FL: 2.2 and 1.7; (3) nonspecific: 0.9 and 0.6; and (4) alternative diagnosis: 0.4 and 0.0. CONCLUSION: Hematoxylin-eosin-stained TBB specimen is of limited usefulness for the diagnosis of FL and should be reserved for patients with intermediate pretest probability of FL. Diffuse LI best discriminates FL from control samples and should be specifically sought. PMID- 9404740 TI - Breathing pattern and arterial blood gases during Nd-YAG laser photoresection of endobronchial lesions under general anesthesia: use of negative pressure ventilation: a preliminary study. AB - STUDY OBJECTIVE: To evaluate the efficacy of negative pressure ventilation (NPV) in avoiding or reducing apneas and related hypoxemia and respiratory acidosis during laser therapy (LT) of endobronchial lesions. DESIGN: A prospective, controlled, randomized study. SETTING: An operating theater of a respiratory endoscopy and laser therapy unit. POPULATION AND INTERVENTION: Twenty-seven consecutive patients referred to LT were entered into the study. Fourteen patients were randomly assigned to LT under general anesthesia and spontaneous assisted ventilation (control group) whereas in 13 cases, NPV by a poncho-wrap ventilator (NPV group) was added to the procedure. MEASUREMENTS AND RESULTS: The prevalence and the duration of apnea/hypopnea periods assessed by respiratory inductive plethysmography during LT were significantly reduced under NPV, compared to the control group. As compared to baseline, during LT, all control patients developed mild to severe hypercapnia (PaCO2 ranging from 55 to 76 mm Hg) and respiratory acidosis (pH from 7.33 to 7.19), whereas only three patients undergoing NPV (23%) developed hypercapnia (PaCO2 from 52 to 68 mm Hg) and related acidosis (pH from 7.29 to 7.21). Optimal oxygenation was achieved in all of the patients; nevertheless, patients under NPV needed a lower mean oxygen supply; five of them (38%) could be treated at a fraction of inspired oxygen of 0.21 for the whole procedure. CONCLUSION: NPV may be useful in reducing apneas during laser therapy under general anesthesia, thus reducing hypercapnia, related acidosis, and need of oxygen supplementation. PMID- 9404741 TI - Cigarette smoking and histologic type of lung cancer in men. AB - STUDY OBJECTIVES: To determine whether intensity, duration, age at initiation, and cessation of cigarette smoking act differently in the development of various histologic types of lung cancer. DESIGN: A case-control study among deceased men who underwent autopsy, a procedure that involves approximately 73% of all local deaths. SETTING: The Province of Trieste in northeastern Italy PARTICIPANTS: Seven hundred fifty-five patients with lung cancer, including 267 with squamous cell carcinoma, 218 with small cell carcinoma, 90 with large cell carcinoma, 158 with adenocarcinoma, and 22 with other histologic types, and 755 control subjects who had died of causes other than chronic lung diseases and certain tumors. Information on smoking habits, residential history, and occupational exposure was obtained from each subject's next of kin. RESULTS: Compared with nonsmokers, the odds ratio (OR) for current smokers was 13.4 for all types combined, 18.8 for squamous cell carcinoma, 14.3 for small cell carcinoma, 34.3 for large cell carcinoma, and 7.9 for adenocarcinoma. Intensity of smoking, duration, age at starting, and dose were all directly associated with all histologic types of lung cancer, although the OR was lower for adenocarcinoma than for other cell types. When results were restricted to ever smokers, exposure-response curves were similar across histologic types. The risk of lung cancer attributable to smoking was 88% for all types combined, 91% for squamous cell carcinoma, 89% for small cell carcinoma, 95% for large cell carcinoma, and 82% for adenocarcinoma. CONCLUSIONS: This study confirms that cigarette smoking causes all types of lung cancer, but the proportion of cases attributable to smoking is lower for adenocarcinoma than for other types, due to a higher proportion of nonsmokers. PMID- 9404742 TI - Mediastinal lymph node staging with FDG-PET scan in patients with potentially operable non-small cell lung cancer: a prospective analysis of 50 cases. Leuven Lung Cancer Group. AB - STUDY OBJECTIVE: To compare the performance of CT, radio-labeled 18F-fluoro-2 deoxy-D-glucose positron emission tomography (FDG-PET) blinded to CT, and FDG-PET visually correlated with CT, in the detection of N2 metastatic mediastinal lymph nodes (MLN) in patients with non-small cell lung cancer (NSCLC) and to hypothesize how PET could influence our actual mediastinal staging procedures. SETTING: Tertiary university hospital. PATIENTS AND METHODS: In 50 patients with potentially operable NSCLC, thoracic CT, PET, and invasive surgical staging were performed. Blinded prospective interpretation was performed for each test and compared with surgical pathology results. Abnormalities on each of these staging examinations were recorded on a standard MLN map. RESULTS: The sensitivity, specificity, and accuracy in detecting N2 disease of CT was 67%, 59%, and 64%, respectively. Results of PET blinded to CT were significantly better (p=0.004): 67%, 97%, and 88%, respectively. For PET visually correlated with CT, this was 93%, 97%, and 96%, respectively. In 22 patients, both CT and PET were normal, and this was correct in all cases. CONCLUSIONS: PET was significantly more accurate than CT in the MLN staging in NSCLC. Both examinations were complementary, since visual correlation with the anatomic information on CT improved the reader's ability to discriminate between hilar vs subaortic MLN FDG uptake, and between paramediastinal tumor vs tracheobronchial MLN FDG uptake. If the results can be confirmed in larger numbers of patients, PET could reduce the need for invasive surgical staging remarkably. PMID- 9404743 TI - Clinical results of treatment of advanced esophageal carcinoma with hyperthermia in combination with chemoradiotherapy. AB - Chemoradiotherapy combined with hyperthermia was administered to 35 patients with advanced esophageal carcinoma who either required preoperative treatment or had nonresectable disease. As a rule, each patient received a total dose of 30 Gy in 15 daily fractions of 2 Gy, 5 d/wk. Bleomycin or cisplatin, in combination with fluorouracil, was employed as chemotherapy. Hyperthermia was applied by intraluminal heating twice a week for a total of six sessions using an apparatus (IH-500T; Japan Crescent Co Ltd; Tokyo, Japan) (radiofrequency, 13.56 MHz) with an intraesophageal applicator and two extracorporeal applicators placed on the chest and back. This treatment method obtained a response rate of 80%, consisting of a complete response rate of 22.9% and partial response of 57.1%. In 15 cases, the tumor became resectable (resectability rate, 42.9%) following treatment. The histologic study of the resected specimens revealed absence of viable tumor cells in five patients (33.3% of the resected cases) (markedly effective), and in six patients (40.0%), the combined therapy was considered to be moderately effective. No complications considered due to hyperthermia itself were recognized. The overall 5-year survival rate was 11.8%. In conclusion, chemoradiotherapy combined with hyperthermia was locally effective, yielding an overall response of 80.0%. However, the prognosis of the patients remains unfavorable. Advanced esophageal carcinoma requires treatment taking into account lymphatic and hematogenic metastasis at the beginning of treatment. PMID- 9404744 TI - Lung volume reduction surgery alters management of pulmonary nodules in patients with severe COPD. AB - OBJECTIVE: To examine the role of lung volume reduction surgery (LVRS) in expanding the treatment options for patients with single pulmonary nodules and emphysema. METHODS: Retrospective review of all patients undergoing LVRS at the University of Michigan between January 1995 and June 1996. Those undergoing simultaneous LVRS and resection of a suspected pulmonary malignancy formed the study group and underwent history and physical examination, pulmonary function tests, chest radiography, and high-resolution CT of the chest. If heterogeneous emphysema was found, cardiac imaging and single-photon emission CT perfusion lung scanning were performed. All study patients participated in pulmonary rehabilitation preoperatively. Age- and sex-matched patients who had undergone standard lobectomy for removal of pulmonary malignancy during the same period formed the control group. RESULTS: Of 75 patients who underwent LVRS, 11 had simultaneous resection of a pulmonary nodule. In 10 patients, the nodules were radiographically apparent with 1 demonstrating central calcification. Histologic evaluation revealed six granulomas, two hamartomas, and three neoplastic lesions (one adenocarcinoma, one squamous cell, and one large cell carcinoma). Preoperative FEV1 was 26.18+/-2.49% predicted in the LVRS group and 81.36+/-6.07% predicted (p=0.000001) in the control group, and the FVC was 65.27+/-5.17% predicted vs 92.18+/-5.53% predicted (p=0.002). Two LVRS patients had a PaCO2 >45 mm Hg while 11 exhibited oxygen desaturation during a 6-min walk test. Postoperative complications occurred in two LVRS patients and three control patients. The mean length of stay in the LVRS group (7.55+/-1.10 days) was not different than in the control group (8.81+/-1.56 days). Three months after LVRS and simultaneous nodule resection, FEV1 rose by 47%, FVC by 25%, and all study patients noted less dyspnea as measured by transitional dyspnea index. CONCLUSIONS: Simultaneous LVRS and resection of a suspected bronchogenic carcinoma is feasible and associated with minimal morbidity and significantly improved pulmonary function and dyspnea. PMID- 9404745 TI - Nebulized wet aerosol treatment in emergency department--is it essential? Comparison with large spacer device for metered-dose inhaler. AB - OBJECTIVE: To determine the efficacy of a metered-dose inhaler (MDI) with a large spacer device as compared to nebulized wet aerosols in the treatment of an unselected population with severe airflow limitation. DESIGN: Randomized, double blind, placebo-controlled trial. SETTING: University Hospital Department of Emergency Medicine (DEM). PATIENTS: Fifty patients, referred to the DEM between October 1, 1994 and March 31, 1995 with a severe, acute obstructive pulmonary event. Thirteen patients were diagnosed as having COPD; 37 patients were diagnosed as having asthma. INTERVENTION AND RESULTS: Patients received either placebo MDI through a 750-mL cone-shaped spacer (Glaxo) [2 puffs] and nebulized salbutamol aerosol 0.5 mL in 1.5 mL saline solution (group 1, n=25) or salbutamol MDI and 0.5 mL saline solution in 1.5 mL saline solution administered in the same manner as above (group 2, n=25). The above treatment was repeated three times every 15 min, unless side effects appeared. Upon enrollment into the study, the FEV1 in group 1 was 0.78+/-0.7 L (mean+/-SD), 32% of predicted, and in group 2, 0.74+/-0.51 L, 29% of predicted (p=0.83). The FEV1 values after the first, second, and third interventions were as follows: in group 1, 1.18+/-0.99 L, 1.40+/-0.8, and 1.47+/-0.79, respectively, and in group 2, 1.17+/-0.99 L, 1.46+/ 1.01, and 1.54+/-0.79 (p=0.83, 0.36, and 0.48, respectively). We observed no difference in spirometric measurements between the two groups at any time. CONCLUSION: Even in the setting of the unselected group of patient referrals to the DEM for episodes of severe airflow limitation, the clinical and the objective bronchodilator responses to the administration of salbutamol are independent of the method of delivery: MDI with a large spacer vs aerosol nebulization. PMID- 9404746 TI - Verbal memory impairment in COPD: its mechanisms and clinical relevance. AB - STUDY OBJECTIVES: Identification of mechanisms accounting for verbal memory impairment in patients with severe COPD; assessing the relationship between verbal memory and the overall cognitive performance; verifying if verbal memory impairment affects medication adherence. DESIGN: Case-comparison study. SETTING: Outpatient Departments of Pneumology and Neurology, Day Hospital of General Surgery. PATIENTS: Forty-two COPD ambulatory patients, age 70+/-9.7 years, with hypoxemia and hypercarbia (group A); 27 normal subjects of comparable age and educational level (group B); 31 patients with Alzheimer's disease (group C); and 26 older normal subjects (group D). MEASUREMENTS AND RESULTS: The overall cognitive function and verbal memory were evaluated by the Mental Deterioration Battery and 14 indexes of verbal memory. Defective retrieval and recognition mechanisms distinguished group A from group B. According to discriminant analysis, verbal memory profile of COPD patients was group specific in 38.1% of cases and conformed to that of group B, C, and D in 19%, 16.7%, and 26.2% of cases, respectively. In COPD patients, both immediate and delayed recall, the strongest determinants of the discriminant function, were significantly correlated with the overall cognitive performance (rho=0.64, p=0.001; rho=0.61, p=0.001, respectively). Poor adherence to medication regimen was significantly associated with abnormal delayed recall score (82.3% vs 36% in subjects with normal delayed recall, p<0.008). CONCLUSIONS: Decline of verbal memory parallels that of the overall cognitive function in COPD patients and is due to the impairment of both active recall and passive recognition of learned material. It could be an important determinant of the level of medication adherence. PMID- 9404747 TI - Routine nebulized ipratropium and albuterol together are better than either alone in COPD. The COMBIVENT Inhalation Solution Study Group. AB - STUDY OBJECTIVE: We compared the long-term safety and efficacy of the combination ipratropium bromide (IB) and albuterol sulfate (ALB) inhalation solution with that of each separate component using three-times-daily administration. DESIGN: Using a parallel design, we randomized patients to receive 3.0 mg ALB, 0.5 mg IB, or the combination by small-volume nebulizer (SVN) for 85 days. Subjects were allowed to use up to two extra doses of study medication daily for control of symptoms on an as-needed basis. The main efficacy evaluation was the acute pulmonary function response to an aerosol of the maintenance study medication over the course of the investigation. Physician global evaluation, subject quality of life assessments, COPD symptom scores, and twice-daily peak expiratory flow rate (PEFR) were also assessed over the study period. SETTING: Twenty-five centers participated in the investigation. PATIENTS: We studied 652 patients with moderate to severe COPD. MEASUREMENTS AND RESULTS: Over the course of the study, the acute spirometric response and evening PEFR values with the SVN combination of IB plus ALB were statistically significantly better compared to ALB or IB alone. The quality of life scores, physician global evaluations, symptom scores, and morning PEFR scores were unchanged over the duration of the study in all treatment groups. There was no significant difference in adverse events in the three treatment groups. CONCLUSIONS: In patients with COPD, maintenance SVN therapy with IB and ALB provides better bronchodilation than either therapy alone without increasing side effects. PMID- 9404748 TI - Evaluation of emphysema in patients with reversible airway obstruction using high resolution CT. AB - OBJECTIVE: This study was carried out to determine whether asthma affects the development of emphysema. METHODS: We studied 62 patients with reversible airway obstruction during remission, and evaluated the presence and severity of emphysema using high-resolution CT. The emphysema score (ES) was evaluated with the visual scoring method on CT scans. RESULTS: Of the 62 patients, 14 were judged to have emphysema. Patients with emphysema were significantly older and more likely to be male than those without emphysema. All patients with emphysema were smokers. There was no significant difference in the duration or severity of asthma between patients with and without emphysema. The 62 patients were divided into three groups according to the ES: 48 patients without emphysema (ES = 0%), 8 patients with mild emphysema (0% < ES < 15%), and 6 patients with more severe emphysema (ES > or = 15%). Highly significant differences between patients without emphysema and those with more severe emphysema were found in FEV1 (p<0.01), FEV1/FVC (p<0.001), diffusing capacity for carbon monoxide (DCO) (p<0.01), and DCO/alveolar volume (p<0.0001). CONCLUSION: Neither the duration nor the severity of asthma was correlated with the presence of emphysema, while smoking history, sex, and age were strongly correlated. No patients with emphysema were found among the nonsmokers, including those with severe asthma or asthma of long duration. These results suggest that asthma does not lead to emphysema. PMID- 9404749 TI - A risk screening questionnaire for adult asthmatics to predict attendance at hospital emergency departments. South Australian Asthma Reference Panel. AB - STUDY OBJECTIVES: To develop a practical screening tool that could identify adult patients highly likely to attend a hospital emergency department (ED) in a 1-year period. DESIGN: Retrospective case-control study of patients who did and did not attend a hospital ED for asthma in the past year. SETTING: Adelaide, South Australia. PARTICIPANTS: One hundred sixty-five adults attending an ED for asthma were compared with 260 adults with asthma from a community survey who had not attended an ED in the previous year. MEASUREMENTS AND RESULTS: The following variables were independently related to ED attendance: having been woken from sleep by asthma in past month; having been admitted to hospital because of asthma in the past year; having seen more than one general practitioner for asthma in the last 12 months; a moderate or severe self-rating of asthma in the last month; and having taken oral steroid medication for asthma in past month. A risk screening questionnaire using the weighted responses to these five variables with a cutoff score of 30/100 demonstrated a sensitivity of 90% and specificity of 88%. CONCLUSIONS: These findings agree with those of previous studies that markers of asthma severity and discontinuity of care are risk factors for adverse asthma outcomes. Validation of the risk screening questionnaire is required in a prospective study. PMID- 9404750 TI - The effect of a peak flow-based action plan in the prevention of exacerbations of asthma. AB - STUDY OBJECTIVE: To determine the effect of a symptom-based and a peak flow-based action plan in preventing acute exacerbations in subjects with poorly controlled asthma. DESIGN: A randomized controlled trial in which subjects who had required urgent treatment for their asthma were allocated to receive no action plan, a symptom-based plan, or a peak flow-based action plan. SETTING: A university hospital asthma clinic. POPULATION: One hundred fifty subjects were recruited after attending an emergency department or a clinic for urgent treatment of asthma. INTERVENTIONS: All subjects received evaluation and education for asthma before being randomly allocated to receive no action plan, a symptom-based action plan, or a peak flowmeter and a peak flow-based action plan. MEASUREMENTS: Subjects were assessed by questionnaire at 3 and 6 months after enrollment with questions relating to their asthma control and their need for urgent treatment or hospital admission for asthma. RESULTS: At 6 months after enrollment, although all three intervention groups experienced improvement in their asthma control, there was a striking reduction in emergency department visits for asthma only in the peak flow-based action plan group (p=0.006). No significant difference in emergency visits was apparent between the symptom-based action plan and no action plan groups. CONCLUSIONS: We conclude that a peak flow-based action plan is effective, at least in the short term, in protecting patients with asthma against severe exacerbations of their disease. PMID- 9404751 TI - Factors that affect normal lung function in white Australian adults. AB - STUDY OBJECTIVE: To classify abnormal lung function in epidemiologic studies, we first calculated "normal" values using data from Australian white adults. We then examined the effects of airway hyperresponsiveness (AHR), respiratory symptoms, current and past asthma, and current smoking on spirometric function. METHODS: A large random sample of 1,527 adults aged 18 to 73 years was studied. We measured respiratory symptoms and smoking history by questionnaire and AHR by histamine inhalation test. RESULTS: Data from 729 "normal" subjects (asymptomatic nonsmokers without AHR) were used to obtain regression models for FVC, FEV1, peak expiratory flow rate, and forced expiratory flow between 25% and 75% of FVC. The R2 values were 0.76, 0.74, 0.58, and 0.29, respectively. The presence of AHR reduced FVC by 0.1 L and FEV1 by 0.2 L, on average. Subjects with asthma-related symptoms had a mean reduction in FVC of 0.1 L for both genders and in FEV1 of 0.08 L for women and 0.2 L for men. Current asthma reduced FVC by 0.3 L, on average, and FEV1 by 0.5 L for women and 0.6 L for men. The FEV1 was reduced by 0.002 L per cigarette smoked daily. CONCLUSION: Recent symptoms, AHR, and current smoking were all important predictors of reduced lung function. PMID- 9404752 TI - Reproducibility of nasal peak inspiratory flow among healthy adults: assessment of epidemiologic utility. AB - STUDY OBJECTIVE: To assess the reproducibility of nasal peak inspiratory flow (PIFn). PARTICIPANTS: Twelve healthy nonsmoking volunteers were studied. METHODS: Repeated measurements of PIFn and oral (PIFm) peak inspiratory flow were performed for 5 consecutive days. Two methods of inhalation were compared. In the residual volume (RV) method, the forced maximal inspiratory maneuver was initiated from the end of a maximal expiration, while in the functional residual capacity (FRC) method, the maneuver was from the end of a tidal breath. Reproducibility was assessed by the intraclass correlation coefficient. Time trend for the 5 days was assessed by random effect models adjusting for different baseline for each subject. RESULTS: The intraclass correlation coefficient (ICC) of PIFn was 0.89 (lower limit of one-sided 95% confidence interval is 0.80) by the RV method and 0.78 (95% lower limit is 0.63) by the FRC method, suggesting that both methods have good reproducibility. These were similar to the ICCs of PIFm by each method. The FRC method did not show a significant time trend for PIFn. The RV method had a small, but significant, decreasing time trend of a magnitude considered inconsequential for the purpose of epidemiologic study. CONCLUSION: PIFn, measured from either RV or from FRC, showed good reproducibility and can be employed in epidemiologic studies investigating the upper airways' response to air pollutant exposure. Further studies of the relationships between PIFn and signs and symptoms of rhinitis are needed to evaluate the utility of this test for clinical and epidemiologic use. PMID- 9404753 TI - Prepubescents' ventilatory responses to exercise with reference to sex and body size. AB - STUDY OBJECTIVES: To examine the ventilatory responses of prepubescent children to submaximal and peak exercise using appropriate allometric modeling to control for differences in body size. DESIGN: Cross-sectional study of a representative sample of children. SETTING: Middle schools (8 to 11 years) in Exeter, UK. PARTICIPANTS: We studied 101 boys and 76 girls aged 11.1 (0.4) years and classified Tanner stage 1 for pubic hair (no true pubic hair). MEASUREMENTS: At rest: stature, mass, sum of skinfolds, hemoglobin concentration, FVC, and FEV1. During treadmill exercise at 7, 8, 9, and 10 km/h, and at peak exercise: oxygen uptake (VO2), minute ventilation (VE), tidal volume (VT), and respiratory frequency (Rf). RESULTS: At peak exercise, boys' VO2, VE, and VT were significantly (p<0.01) higher than girls' values and remained so even when the influence of body size was controlled using allometric principles. There were no significant (p>0.05) sex differences in Rf or the ratios VT/FVC or VE/VO2. When data were compared at the same relative exercise intensity (ie, 70 to 75% or 80 to 85% peak VO2), no significant (p>0.05) sex differences in Rf, VT/FVC, or VE/VO2 were detected. Boys' higher (p<0.001) VO2 values were reflected by their higher VE which remained higher than values for girls at both submaximal levels even when the influence of body size was covaried out. CONCLUSIONS: Prepubescent boys demonstrate higher peak VO2 than girls and this is supported by a higher VE and VT, even when the influence of body size is accounted for using allometry. Other ventilatory responses to both peak exercise and exercise at the same relative intensity are remarkably similar in both boys and girls. PMID- 9404754 TI - Accidents in obstructive sleep apnea patients treated with nasal continuous positive airway pressure: a prospective study. The Working Group ANTADIR, Paris and CRESGE, Lille, France. Association Nationale de Traitement a Domicile des Insuffisants Respiratoires. AB - Many studies have shown a relationship between obstructive sleep apnea (OSA) and accidents, but to our knowledge, none have investigated prospectively the effects of treatment with nasal continuous positive airway pressure (CPAP). CPAP was proposed to 973 patients, of whom 893 patients actually underwent CPAP. These patients were consecutively invited to enter a prospective follow-up study including a questionnaire before treatment and after 6 and 12 months of treatment; 547 patients completed the study (153 left the study, and only partial data were available for 193). The baseline questionnaire included questions concerning accidents in the previous 12 months, asking whether patients had had an accident and, if so, whether they felt that the accident(s) were related to sleepiness, and whether the patients felt that they had had near-miss accidents due to sleepiness. The questionnaires at 6 and 12 months included the same questions referring to the previous 6 months; the accidents reported on each follow-up questionnaire were cumulated and compared with the accidents during the 1-year period before treatment. The number of patients having an accident decreased with treatment for real accidents (from 60 to 36; p<0.01), as well as for near-miss accidents (from 151 to 32; p<0.01). The average number of accidents per patient also decreased, for real accidents (from 1.6+/-1.3 to 1.1+/-0.3; p<0.01) and for near-miss accidents (from 4.5+/-6.5 to 1.8+/-1.4; p<0.01). The cost, in terms of days in hospital related to accidents, decreased from 885 to 84 days. With caution due to the absence of a control group, it is suggested that treatment with CPAP decreases the number of accidents occurring in OSA patients. This result may have important implications in the evaluation of the cost/benefit ratio when treating OSA patients. PMID- 9404755 TI - Respiratory and arousal responses to acoustic stimulation. AB - STUDY OBJECTIVES: Although sleep-related obstructive apnea is most often associated with transient arousal, the impact of this arousal on respiratory control remains unclear. We tested the hypotheses that acoustic arousing stimulation can generate a significant respiratory response during sleep in healthy subjects and that the magnitude or timing of this response is affected by the presence of electrocortical arousal or inhaled carbon dioxide. DESIGN: We employed binaural tone bursts (0.5-s duration, 4-KHz center frequency, 99-s interstimulus interval) to elicit repetitive transient arousals from sleep during nocturnal polysomnographic recordings beginning at 10 PM and ending at 6 AM. PARTICIPANTS: Recordings were conducted in five healthy adult volunteers aged 24 to 37 years. INTERVENTIONS: Inspired gas was alternated between room air and 3% to 7% CO2 (titrated to yield an approximate 50% increase in minute ventilation) at 1-h intervals. MEASUREMENTS AND RESULTS: Each 30-s epoch was scored for sleep/wake stage according to standard criteria. Only results obtained during nonrapid eye movement sleep are presented herein. Tone-evoked arousals were detected by computer analysis as increased EEG frequency occurring within 3 s of acoustic stimulation. For each tone, respiratory parameters for each of three prestimulus and four poststimulus breaths were normalized to the overall mean of prestimulus breaths measured during room air breathing for each subject. Tone bursts elicited repetitive transient arousals with a mean duration of approximately 10 s from all stages of sleep. With respect to the three prestimulus breaths, acoustic stimulation was associated with increased tidal volume and decreased inspiratory duration for at least four breaths. These respiratory responses to acoustic stimulation were not significantly influenced by either presence of transient arousal from sleep or inspired gas. CONCLUSIONS: We conclude that transient EEG arousal may be repeatedly evoked from nonrapid eye movement sleep by transient acoustic stimulation in normal sleepers. This sensory stimulation is associated with augmented ventilation, a response that is not significantly affected by inspired hypercapnia or the presence of generalized EEG arousal. PMID- 9404756 TI - Endothelial selectins in acute mountain sickness and high-altitude pulmonary edema. AB - STUDY OBJECTIVES: Mechanical or inflammatory injury to pulmonary endothelial cells may cause impaired pulmonary gas exchange in acute mountain sickness (AMS) and noncardiogenic pulmonary edema in high-altitude pulmonary edema (HAPE). This study was designed to determine whether markers of endothelial cell activation or injury, plasma E- and P-selectin, were increased after ascent to high altitude, in AMS or in HAPE. DESIGN: We collected clinical data and plasma specimens in control subjects at sea level and after ascent to 4,200 m, and in climbers with AMS or HAPE at 4,200 m. Data analysis was performed using standard nonparametric statistical methods, and results reported as mean+/-SD. SETTING: National Park Service medical camp at 4,200 m on Mt. McKinley (Denali), Alaska. PATIENTS: Blood samples and clinical data were collected from 17 healthy climbers at sea level and again after ascent to 4,200 m, and from a different group of 13 climbers with AMS and 8 climbers with HAPE at 4,200 m. Climbers with AMS were divided into normoxic (n=7) and hypoxemic (n=6) groups. MEASUREMENTS AND RESULTS: Using an enzyme immunoassay technique, plasma E-selectin concentrations were found to be increased in the 17 control subjects after ascent to 4,200 m (17.2+/-8.2 ng/mL) as compared to sea level (12.9+/-8.2 ng/mL) (p=0.001). Plasma E-selectin concentrations were also increased in subjects with hypoxemic AMS (30.6+/-13.4 ng/mL) and HAPE (23.3+/-9.1 ng/mL) compared to control subjects at sea level (p=0.009). Increased plasma E-selectin concentration significantly correlated with hypoxemia (p=0.006). Plasma P-selectin concentrations were unchanged after ascent to 4,200 m and in subjects with AMS and HAPE. CONCLUSION: Because E selectin is produced only by endothelial cells, increased plasma E-selectin after ascent to high altitude and in hypoxemic climbers with AMS and HAPE provides evidence that endothelial cell activation or injury is a component of hypoxic altitude illness. PMID- 9404757 TI - Urokinase in the management of complicated parapneumonic effusions in children. AB - OBJECTIVE: Use of intrapleural fibrinolytic agents in the management of complicated parapneumonic effusions has been widely reported in adults. Such agents promote drainage of fluid through the thoracostomy tube and may obviate surgery. Both streptokinase and urokinase have been used for this purpose, but there are few reports of their use in the children. The objective of this study was to evaluate the role of intrapleural urokinase in the management of complicated parapneumonic effusions in children. METHODS: We reviewed the hospital course of nine children, ages 6 months to 6 years, with complicated parapneumonic effusions who received intrapleural urokinase after failing to respond to I.V. antibiotics and closed-tube thoracostomy drainage. Four subjects had additional thoroscopic adhesiolysis before intrapleural instillation of urokinase; 20,000 IU of diluted urokinase was instilled three times a day via the thoracostomy tube for 3 days. RESULTS: Eight subjects responded to 3 days of urokinase instillation, with increased thoracostomy tube drainage and clinical resolution of symptoms. The remaining subject responded to a second course of instillation. Two subjects needed oral analgesic for transient chest pain. All subjects tolerated the procedure well. No bleeding, fever, anaphylaxis, or allergic reactions were noted. The coagulation parameters remained unchanged. CONCLUSION: Intrapleural instillation of urokinase appears to be a useful and safe adjunct in the management of complicated parapneumonic effusions in children. Its use may be considered in potential decortication patients in an effort to prevent surgery and possibly shorten hospitalization. PMID- 9404758 TI - Effects of early defibrillation by ambulance personnel on short- and long-term outcome of cardiac arrest survival: the Munich experiment. AB - OBJECTIVES: This study evaluates the feasibility of implementing early defibrillation of out-of-hospital cardiac arrest patients for basic life-support providers (EMT-D) in a two-tier emergency system in the city of Munich, Germany. DESIGN: Retrospective consecutive analysis of all EMT-D attempts during a 5-year initiation phase (1990 to 1994) and prospective follow-up of all cardiac arrest survivors discharged from hospital. SETTING: A strictly defined inner-city and suburban area of 978 km2 and a residential population of 1,530,000 inhabitants with 22 ICUs in urban hospitals. One dispatching center to alert a two-tier emergency system with 56 EMT-D-staffed ambulances and physician-staffed mobile ICUs stationed at the nearest of nine hospitals. METHODS: AH EMT-D cases were identified and data on patients were documented in a standardized manner from patients' records, including the resuscitation protocol in the hospitals to which the patients were referred. For those patients discharged from the hospital, a standardized telephone interview was undertaken with the physician in charge of the patient and with the patient/relative leading to an assessment of the patient's status according to the Glasgow-Pittsburgh cerebral performance categories. INTERVENTION: None. RESULTS: During the 5-year initiation phase of the EMT-D program in the two-tier emergency system in Munich, there were 243 resuscitation attempts by EMTs, using the semiautomated defibrillator; 125 patients died immediately on the scene. In 118 patients, spontaneous circulation was reestablished and these patients were admitted to an ICU in 1 of the 22 urban hospitals. Median call-response interval for the EMT-D was 5 min (interquartile range, 3 to 6) and was 10 min (interquartile range, 7 to 13) for the second tier (p < or = 0.0001). In 34 cases (28.8%), EMT-D staff had reestablished spontaneous circulation (ROSC) before the second tier arrived on the scene. Patients with ROSC on the arrival of the second tier were more frequently discharged alive from hospital than were patients without ROSC at that time (p < or = 0.0001). The hospital discharge rate of initially successful resuscitated patients presenting with out-of-hospital ventricular fibrillation was 38.1% (45/118). Overall success rate of all EMT-D attempts was 18.5% (45/243). After a mean follow-up time of 39 (range, 22 to 64) months, 29 (66%) patients were still living. Twenty-five (56.8%) were neurologically not disabled or mildly disabled (CPC 1/2); disability was moderate in 3 (6.8%) patients and was severe in 1 (2.3%) patient. One case was lost to follow-up. CONCLUSION: The present study demonstrates that the upgrading of basic life support providers with semiautomated defibrillators has a significant benefit for cardiac arrest victims outside the hospital in an urban environment. PMID- 9404759 TI - Patient-ventilator trigger asynchrony in prolonged mechanical ventilation. AB - STUDY OBJECTIVE: To investigate patient-ventilator trigger asynchrony (TA), its prevalence, physiologic basis, and clinical implications in patients requiring prolonged mechanical ventilation (PMV). STUDY DESIGN: Descriptive and prospective cohort study. SETTING: Barlow Respiratory Hospital (BRH), a regional weaning center. PATIENTS: Two hundred consecutive ventilator-dependent patients, transferred to BRH over an 18-month period for attempted weaning from PMV. METHODS AND INTERVENTIONS: Patients were assessed clinically for TA within the first week of hospital admission, or once they were in hemodynamically stable condition, by observation of uncoupling of accessory respiratory muscle efforts and onset of machine breaths. Patients were excluded if they had weaned by the time of assessment or if they never achieved hemodynamic stability. Ventilator mode was patient triggered, flow control, volume cycled, with a tidal volume of 7 to 10 mL/kg. Esophageal pressure (Peso), airway-opening pressure, and airflow were measured in patients with TA who consented to esophageal catheter insertion. Attempts to decrease TA in each patient included application of positive end expiratory pressure (PEEP) stepwise to 10 cm H2O, flow triggering, and reduction of ventilator support in pressure support (PS) mode. Patients were followed up until hospital discharge, when outcomes were scored as weaned (defined as >7 days of ventilator independence), failed to wean, or died. RESULTS: Of the 200 patients screened, 26 were excluded and 19 were found to have TA. Patients with TA were older, carried the diagnosis of COPD more frequently, and had more severe hypercapnia than their counterparts without TA. Only 3 of 19 patients (16%), all with intermittent TA, weaned from mechanical ventilation, after 70, 72, and 108 days, respectively. This is in contrast to a weaning success rate of 57%, with a median (range) time to wean of 33 (3 to 182) days in patients without TA. Observation of uncoupling of accessory respiratory muscle movement and onset of machine breaths was accurate in identifying patients with TA, which was confirmed in all seven patients consenting to Peso monitoring. TA appeared to result from high auto-PEEP and severe pump failure. Adjusting trigger sensitivity and application of flow triggering were unsuccessful in eliminating TA; external PEEP improved but rarely led to elimination of TA that was transient in duration. Reduction of ventilator support in PS mode, with resultant increased respiratory pump output and lower tidal volumes, uniformly succeeded in eliminating TA. However, this approach imposed a fatiguing load on the respiratory muscles and was poorly tolerated. CONCLUSION: TA can be easily identified clinically, and when it occurs in the patient in stable condition with PMV, is associated with poor outcome. PMID- 9404760 TI - The influence of high-frequency jet ventilation with varying cardiac-cycle specific synchronization on cardiac output in ARDS. AB - BACKGROUND: Previous studies have shown "beat-to-beat" variation in systemic BP with high-frequency jet ventilation (HFJV). However, it is not clear if such changes are paralleled by changes in cardiac output. OBJECTIVE: To characterize the effect of HFJV near or equal to the heart rate (HR) on beat-to-beat cardiac output in an adult human subject with ARDS. DESIGN: Case study. SETTING: ICU, university teaching hospital. PATIENTS: One patient with end-stage liver disease complicated by sepsis, severe pancreatitis, ARDS, and multisystem organ failure. METHODS: The patient was intubated, sedated, paralyzed, and ventilated with controlled mechanical ventilation (CMV). Ventilatory mode was then switched to HFJV at fixed frequencies (f) near but not equal to the HR (f= 100, 110, and 120 beats/min; HR=108/min). HFJV was then synchronized to the ECG such that f and HR were equal. Continuous cardiac output (COc) was monitored during change of ventilator mode from CMV to fixed-rate HFJV to synchronized HFJV, then followed through progressive delays in jet triggering within the cardiac cycle during the synchronous HFJV mode. COc was monitored by arterial pulse-contour analysis, allowing assessment of beat-to-beat changes in cardiac output. MEASUREMENTS AND MAIN RESULTS: A cyclic variation in COc equal to the beat frequency difference between f and HR was observed (harmonic interaction) during fixed-rate HFJV. This COc oscillation was abolished during synchronous HFJV. COc was significantly greater during systolic synchronous HFJV as compared to diastolic synchronous HFJV or fixed-rate HFJV (10.1 to 9.0 [p<0.05] and to 8.6 [p<0.05] L/min, systolic synchronous to diastolic synchronous and to fixed-rate HFJV, respectively). CONCLUSIONS: This study demonstrates instantaneous variations in cardiac output in a human subject with fixed rates of HFJV near to the HR in humans. These variations are abolished by synchronous HFJV but cardiac output was dependent on the timing of the HFJV inspiration in relation to the cardiac cycle. COc is a potentially valuable method to monitor sudden changes in cardiac output and facilitate attempts to maximize cardiac output during synchronized HFJV. PMID- 9404761 TI - Hyperoxia exacerbates microvascular lung injury following acid aspiration. AB - OBJECTIVES: To examine the effects of an increase in ambient oxygen (O2) concentrations on the extent of inflammatory pulmonary damage following acid aspiration. DESIGN: Prospective, controlled laboratory study. SETTINGS: University-affiliated animal research facility. SUBJECTS: Male, Long Evans rats weighing 250 to 300 g. INTERVENTION: Rats were injured by instillation of 1.2 mL/kg normal saline solution/HCl, pH= 1.25 (acid), into the lungs via a tracheotomy. Animals were allowed to awaken and were exposed to 21%, 50%, or 98% O2 for 0 to 5 h (n/group > or = 10). In a separate set of experiments, injured rats exposed to 98% O2 were treated with different doses of deferoxamine, just prior to injury. Uninjured rats and rats injured with normal saline solution, pH = 5.3, were used as the control group. MEASUREMENTS: Injury was determined by assessing lung function (lung compliance and arterial blood gases) and alveolar capillary wall integrity (wet/dry weight, lung albumin permeability index [PI], and intrapulmonary hemorrhage [HI]). RESULTS: Intrapulmonary instillation of acid increased PI, HI, and decreased static lung compliance compared to uninjured control animals. Increased ambient oxygen following acid aspiration decreased lung compliance, 1.06+/-0.03 mL/kg/cm H2O, in oxygen-exposed lungs when compared to the lungs exposed to air, 1.26+/-0.04, following a low pH aspirate (p<0.05). An increase in protein leakage into the lung tissue was noted in oxygen-exposed animals, PI=1.33+/-0.10, vs air-exposed rats, 0.89+/-0.07 (p<0.05). The hyperoxia induced increase in lung injury was prevented by 30 mg/kg or higher deferoxamine treatment, 0.78+/-0.05 (p<0.05). Exposure of animals to 98% O2 for 2 h was sufficient to produce the same increase in microvascular protein leakage as 5-h exposure to O2 following low pH aspirate. CONCLUSION: Hyperoxia increases acid aspiration-induced inflammatory microvascular lung injury. This appears to be mediated by production of reactive species of O2. PMID- 9404762 TI - Elevated levels of soluble adhesion molecules in serum of patients with diffuse panbronchiolitis. AB - STUDY OBJECTIVE: Adhesion molecules have been implicated in the pathogenesis of inflammatory diseases. This study was designed to determine whether soluble adhesion molecules in serum reflect the disease activity in diffuse panbronchiolitis (DPB). PATIENTS AND METHODS: Using an enzyme-linked immunosorbent assay, we measured the serum levels of soluble L-, E-, and P selectin (sL-, sE-, and sP-selectin), intercellular adhesion molecule-1, and vascular cell adhesion molecule-1 in 27 patients with DPB, 13 with bronchiectasis, and 15 normal adults. BAL was also performed, and the levels of interleukin (IL)-8 and IL-1 beta in BAL fluid (BALF) were measured. RESULTS: The serum levels of these molecules were significantly elevated in DPB patients compared with the control subjects. DPB patients also had significant high levels of circulating sE- and sP-selectin compared with patients with bronchiectasis. There was a significant correlation between serum sE-selectin and the percentage of neutrophils in BALF in all patients. There was a significant inverse correlation between serum sE-selectin and percent vital capacity in DPB patients. In the same patients, the relationships between serum sE-selectin and BALF concentrations of IL-1 beta as well as between serum sL-selectin and BALF IL-8 were also significant. Treatment of DPB patients with macrolides significantly reduced the serum levels of these soluble adhesion molecules and BALF concentrations of IL-1 beta and IL-8. CONCLUSIONS: Our results suggest that these soluble adhesion molecules, particularly selectins, may reflect the disease activity of DPB, and that their levels may be regulated by cytokines produced in the lungs. PMID- 9404763 TI - Pulmonary vascular impedance and recipient chronic pulmonary hypertension following cardiac transplantation. AB - STUDY OBJECTIVES: Recipient chronic pulmonary hypertension (CPH), secondary to long-standing congestive heart failure, represents a significant risk factor for right ventricular (RV) dysfunction following orthotopic cardiac transplantation (TX). This study was designed to characterize the changes occurring in pulmonary hemodynamics, pre-TX and post-TX, using Fourier analysis, a canine model of bicaval TX, and monocrotaline pyrrole (MCTP)-induced recipient CPH. DESIGN: Prospective, controlled study. SETTING: Experimental laboratory. PARTICIPANTS: Twenty adult male mongrel dogs (23 to 26 kg). INTERVENTIONS: Recipients underwent pulmonary artery injection of 3 mg/kg MCTP 4 months pre-TX. On the day of TX, donor hearts were instrumented with an ultrasonic flow probe and micromanometers. Harmonic derivation of functional data was achieved with Fourier analysis. MEASUREMENTS AND RESULTS: At the time of TX, significant increases were observed in the mean pulmonary artery pressure and pulmonary vascular resistance of recipient animals in comparison to donors, which were further significantly increased following the termination of cardiopulmonary bypass. Significant increases were also observed in the input resistance, characteristic impedance, and RV hydraulic power post-TX compared to pre-TX, and occurred in association with a significant decrease in the transpulmonary efficiency. CONCLUSIONS: In the setting of MCTP-induced recipient CPH donor hearts were exposed to significant alterations in cardiopulmonary hemodynamics following bicaval TX. Pulmonary blood flow is maintained by a significantly higher energy expenditure by the RV, but at a lower level of efficiency. This experimental model may provide a useful means by which to evaluate therapeutic options to better manage cardiopulmonary hemodynamics in order to prevent RV failure following TX in the setting of recipient CPH. PMID- 9404764 TI - Disease management of COPD with pulmonary rehabilitation. AB - Pulmonary rehabilitation is a set of tools and disciplines that attends to the multiple needs of the COPD patient. It extends beyond standard care by addressing the disabling features of chronic and progressive lung disease. It centers on self-management, exercise, functional training, psychosocial skills, and contributes to the optimization of medical management. Exercise enables other components by building strength, endurance, confidence, and reducing dyspnea. Patients who have undergone rehabilitation often enjoy a reduced need for health care utilization. On the downside, rehabilitation is a one-time intervention, the benefits of which dissolve over time. The patient's physician is rarely a participant in the program; thus, the physician is at a disadvantage in being able to support a long-term response. Rehabilitation is available to a small percentage of a large patient population who could benefit. Optimal disease management would entail redesigning standard medical care to integrate rehabilitative elements into a system of patient self-management and regular exercise. It should emphasize physician involvement in self-management, which is essential in developing and maintaining an effective exacerbation protocol. Pulmonary rehabilitation should take its place in the mainstream of disease management through its integrative and reconciliative role in the multidisciplinary continuum of services, as defined by the National Institutes of Health, Pulmonary Rehabilitation Research, Workshop of 1994. PMID- 9404765 TI - Penicillin dosing for pneumococcal pneumonia. AB - Most textbook authors still endorse penicillin G as the specific antibiotic of choice for pneumococcal pneumonia. However, problems with early precise etiologic diagnosis of pneumonia and the emergence of drug-resistant pneumococci cause penicillin to be seldom used for this purpose today. A third explanation for the infrequent use of penicillin is lack of clear consensus dosing guidelines. Emergence of pneumococci resistant to the newer cephalosporins and concerns about overuse of vancomycin, however, have prompted renewed interest in the development of precise, rapid methods for diagnosis of pneumococcal pneumonia with the implication that penicillin might be used more frequently. We review several issues concerning penicillin dosing: intermittent vs continuous therapy, high dose vs low dose, relationship of dose to resistance, and cost-effective pharmacology. An optimum "high-dose" regimen for life-threatening pneumococcal pneumonia in a 70-kg adult consists of a 3 million unit (mu) loading dose followed by continuous infusion of 10 to 12 mu of freshly prepared drug every 12 h. The maintenance dose should be reduced in elderly patients and in patients with renal failure according to the following formula: dose (mu/24 h = 4+[creatinine clearance divided by 7]). This regimen provides a penicillin serum level of 16 to 20 microg/mL, which should suffice for all but the most highly resistant strains (minimum inhibitory concentration > or = 4 microg/mL). Newer cephalosporins and vancomycin can be reserved for patients with suspected meningitis or endocarditis or for localities in which highly resistant pneumococci are known to be prevalent. PMID- 9404766 TI - Nebulized lidocaine administered to infants and children undergoing flexible bronchoscopy. AB - STUDY OBJECTIVES: The purpose of this study was to evaluate the safety and efficacy of nebulized lidocaine hydrochloride as a topical anesthetic for use during flexible bronchoscopy in infants and children. DESIGN: This was a prospective, randomized, double-blind study. PATIENTS: Twenty consecutive patients scheduled for flexible bronchoscopy who were not intubated and had no known cardiac or hepatic disease comprised the study group. INTERVENTIONS: The patients were randomized to receive either 8 mg/kg or 4 mg/kg of nebulized 2% lidocaine by face mask prior to bronchoscopy. SETTING: The study took place in a bronchoscopy suite at an academic medical center. MEASUREMENTS: To determine systemic absorption, serum lidocaine levels were obtained. To assess efficacy of nebulized lidocaine as a topical anesthetic, changes in heart rate and blood pressure were recorded, and the bronchoscopist (who did not know the lidocaine dose used) rated the ease of passage of the bronchoscope through nose, vocal cords, trachea, bronchi, and all sites overall, and the degree of cough. RESULTS: Nebulized lidocaine was safe, was well-tolerated, and provided adequate anesthesia for half of the patients. The serum lidocaine levels were much lower than the levels in the toxic range. There was a trend toward easier passage of the bronchoscope in the high-dose group at all sites noted previously that were evaluated. CONCLUSION: Nebulized lidocaine in doses up to 8 mg/kg appears to be safe and moderately effective as a topical anesthetic for flexible bronchoscopy in infants and children. The serum levels were remarkably low. Fifty percent of the subjects required no supplemental lidocaine. PMID- 9404768 TI - Shock in a patient exposed to rodents. PMID- 9404769 TI - Two asymptomatic patients with mediastinal disease. PMID- 9404767 TI - Opening of infectious giant bulla with use of video-assisted thoracoscopic surgery. AB - In 5 cases, an infectious giant bulla was opened with the use of video-assisted thoracoscopic surgery (VATS). Because all bullae adhered to the thoracic wall and were noncommunicating with the airway, they were opened without complete resection, leaving their inside walls at the lung and lateral walls on the thoracic wall. The expansion of remnant lung was excellent, and postoperative air leakage did not occur in any case. The postoperative vital capacity and FEV1 improved significantly over the preoperative condition (p<0.01). Because the bronchial communication of bulla is frequently obliterated after infection within the bulla, opening of a bulla is curative and simpler, more effective, and less invasive than complete resection. PMID- 9404770 TI - Retrocardiac mass in a patient with cirrhosis. PMID- 9404771 TI - A rare cause of dyspnea and arterial hypoxemia. AB - Platypnea-orthodeoxia is a rare pattern of dyspnea with arterial hypoxemia. Platypnea is defined as dyspnea induced by upright posture, and it is relieved by the recumbent position. Orthodeoxia refers to arterial desaturation resulting from assuming an erect or upright position. The case reported involves a 59-year old man with profound, unexplained dyspnea despite extensive investigation performed at the referring institution. The difficulty in diagnosis persisted until it was recognized that the investigations, in having been performed under "standard" (supine) conditions, were insufficient and therefore misleading. Despite normal supine intracardiac pressures, a patent foramen ovale was shown to give rise to a large orthostatic intracardiac shunt, demonstrated by means of an echocardiogram performed with the patient supine and upright. Surgical closure of the foramen was followed by dramatic clinical improvement. Among dyspneic patients, discernment of a pattern of platypnea and orthodeoxia is key to effective evaluation. PMID- 9404772 TI - Platypnea-orthodeoxia related to aortic elongation. AB - An 80-year-old woman presented with progressive shortness of breath. There was no history of pulmonary or cardiac disease. Results of a physical examination were normal. She had significant oxygen desaturation while she was in an upright position. Admission to the hospital for workup followed, and evaluation included tilt-table transesophageal echocardiogram and cardiac catheterization. A massive right-to-left shunt through a patent foramen ovale was detected, and surgical intervention resulted in dramatic improvement of symptoms. In this patient, it seems that the syndrome of platypnea-orthodeoxia was related to aortic elongation, allowing significant right-to-left shunt. PMID- 9404773 TI - Intratracheal ectopic thyroid tissue mass. AB - A diagnosis of severe stridor due to a subglottic tracheal mass was made in a 62 year-old woman. The tumor was removed by external transtracheal surgery, and the pathologic study disclosed that it was intratracheal ectopic thyroid tissue. In this particular case, 10 years prior to the onset of the stridor, normal thyroid tissue was seen in biopsy specimens of the same subglottic localization. Although the pathogenesis of a mass composed of intratracheal thyroid tissue is not known, this case shows that it can be a slowly progressive tumor that is asymptomatic for several years. PMID- 9404774 TI - End-stage cystic fibrosis: improved diabetes control 2 years after successful isolated pancreatic cell and double-lung transplantation. AB - Over a period of years, insulin-dependent diabetes and respiratory insufficiency developed in a 35-year-old patient with end-stage cystic fibrosis. After waiting more than 4 years while receiving maintenance treatment with continuous liquid O2 and nasal ventilation, the patient underwent double-lung and pancreatic islet cell transplantation. Subsequently, the patient has enjoyed a normal life with full employment and much better control of his diabetes. Pancreatic islet cell transplantation is a simple and innocuous technique easily added to the end of lung transplantation. These new pancreatic cells, although locally injected, are still secreting more than 2 years later as assessed by repeated C-peptide measurements. PMID- 9404775 TI - Pulmonary cholesterol crystal embolization. AB - BACKGROUND: Cholesterol crystal embolization (CCE) has been documented to affect nearly every organ system. However, CCE involving the lung is distinctly uncommon and has been documented only in the setting of an aortocaval fistula. DESIGN: A case at the Massachusetts General Hospital and a MEDLINE search of English language medical articles published between 1966 and 1997 provide the basis for this report. RESULTS: The precipitants of CCE include invasive vascular procedures, anticoagulant therapy, and thrombolysis. The most common symptoms include claudication of the calf, gastrointestinal bleeding, and weight loss. The most common signs include livedo reticularis, gangrene, and ulcers. Azotemia, proteinuria, normocytic anemia, and eosinophilia often are found. Herein is described the first pathologically confirmed case of CCE to the lung in the absence of an arteriovenous fistula. CONCLUSION: Pulmonary hemorrhage should now be included in the diverse list of presenting signs of CCE. Moreover, CCE should be considered in the differential diagnosis of pulmonary-renal syndromes. PMID- 9404777 TI - Diverse presentation of aberrant origin of the right subclavian artery: two case reports. AB - Aberrant origin of the right subclavian artery occurs in up to 1% of the population and can result in a wide range of symptoms. In this report, two cases of this anomaly are presented. In the first case, a patient developed fatal group A streptococcal aortitis. In the second case, the patient complained of chronic cough and intermittent dyspnea. The embryologic genesis of this abnormality is discussed and the current literature is summarized. Although relatively uncommon, it is important to consider this vascular anomaly in the differential diagnosis of patients with dysphagia, dyspnea, chest pain, fever, or mediastinal widening evidenced on chest roentgenography. PMID- 9404776 TI - Acute life-threatening toxocaral tamponade. AB - An unusual case of life-threatening visceral larva migrans (toxocariasis) is reported herein. The patient was admitted with acute dyspnea and bilateral pleural effusion; rapidly pericardial tamponade developed. Blood and body fluid eosinophilia were elevated. Extensive investigations revealed no malignant process or vasculitis, but Toxocara infection was confirmed by rising specific antibody titers. The high seroprevalence of Toxocara antibodies, particularly in children, suggests that a diagnosis of visceral larva migrans should be considered before a diagnosis of systemic hypereosinophilic syndrome even when clinical presentation is unusual. Prophylaxis against this widespread polymorphic zoonotic infection is desirable in view of the potentially dramatic consequences of infestation. PMID- 9404778 TI - Bronchiolitis obliterans with organizing pneumonia and cold agglutinin disease associated with phenytoin hypersensitivity syndrome. AB - Phenytoin hypersensitivity syndrome (PHS) is a rare delayed hypersensitivity reaction which occurs following exposure to phenytoin sodium. Pulmonary involvement is uncommonly described. Herein is reported the first case of histopathologic bronchiolitis obliterans organizing pneumonia (BOOP) found on open-lung biopsy in a patient with severe PHS. New onset, clinically significant, cold agglutinin disease was also documented. Hemodynamic parameters mimicking sepsis were present in the absence of significant clinical infection. Rapid, dramatic improvement followed high-dose steroid therapy. PMID- 9404780 TI - Treatment of diffuse tracheomalacia secondary to relapsing polychondritis with continuous positive airway pressure. AB - Relapsing polychondritis (RP) is a rare disease characterized by recurrent inflammation and destruction of the cartilaginous structures. Tracheobronchial chondritis is a dreaded complication of RP. We wish to report a case of RP of the trachea and bronchi which was treated with nasal continuous positive airway pressure. PMID- 9404779 TI - Vasculitis and bronchiectasis in a patient with antibodies to bactericidal/permeability-increasing protein and alpha1-antitrypsin deficiency. AB - A patient with alpha1-antitrypsin deficiency is reported herein; this subject developed aggressive bronchial disease and recurrent cutaneous vasculitis after pulmonary infection with Pseudomonas aeruginosa. Autoantibodies to neutrophil cytoplasmic antigens were detected, which produced granular cytoplasmic staining by indirect immunofluorescence with specificity for a newly characterized antigen: bactericidal/permeability-increasing protein (BPI). The bronchial disease and vasculitis improved, and the IgA anti-BPI titer fell after antipseudomonal treatment. This raises the possibility that anti-BPI antibodies contributed to both the bronchial disease and vasculitis. PMID- 9404781 TI - Necrotizing tracheobronchitis with progressive airflow obstruction associated with paraneoplastic pemphigus. AB - Paraneoplastic pemphigus (PNP) is an autoimmune disease associated with leukemia and non-Hodgkin's lymphoma. A patient with stage IVB poorly differentiated lymphocytic lymphoma developed characteristic upper and lower airway involvement with profound mucocutaneous erosion and tracheobronchial epithelial desquamation. Immunofluorescence testing confirmed autoantibody deposition along the basement membrane of bronchial epithelium. Disruption of the cellular adhesion mechanisms, including desmosomes, hemidesmosomes, and possibly the integrin subunits, is presumed to have led to disruption and desquamation of the tracheobronchial epithelial barrier, severe obstruction of the airways and hypoxia, and possibly bacterial superinfection. As far as can be determined, the feature of airflow obstruction occurring in association with PNP has not been described. Physicians should be aware that these complications of PNP may rapidly lead to hypoxic respiratory failure and death. PMID- 9404782 TI - Antiphospholipid antibody syndrome presenting as a refractory noninflammatory pulmonary vasculopathy. AB - The clinical manifestations of antiphospholipid antibody syndrome (APLAS) are protean. Pulmonary manifestations are often thromboembolic in origin; ARDS and pulmonary hypertension have been reported as features of a widespread vasculopathy associated with systemic lupus or Sjogren's syndrome. This is the report of a woman with primary APLAS who died of a noninflammatory pulmonary vasculopathy. The case is unusual in its pulmonary manifestations, its initial response to corticosteroids and antithrombotic medications, its failure to stabilize with high-intensity warfarin sodium and aspirin treatment, and finally its fulminant progression despite multiple interventions. PMID- 9404783 TI - www.journal--good or bad? PMID- 9404784 TI - CyberCHEST. PMID- 9404785 TI - The power of electronic publishing: enlightened and converted. PMID- 9404786 TI - A complete World Wide Web without CHEST? PMID- 9404787 TI - Pneumonitis due to Mycobacterium avium complex in hot tub water: infection or hypersensitivity? PMID- 9404788 TI - Pulmonary vascular involvement in pulmonary histiocytosis X. PMID- 9404789 TI - Usefulness of walking test for arterial oxygen desaturation screening in general population. PMID- 9404790 TI - Additional experiences with corticosteroids in COPD. PMID- 9404791 TI - The evaluation of eye patching in the treatment of traumatic corneal epithelial defects. AB - The objective of this study was to compare the rates of healing for patched and non-patched traumatic corneal epithelial defects (CEDs) after 1 day of treatment. To achieve this we initiated a randomized, controlled, prospective, clinical investigation comparing patching vs. non-patching of CEDs. Patients were evaluated initially and at 24 h using slit lamp biomicroscopy, and each corneal epithelial defect was documented on standardized initial and follow-up grid sheets. Percentage of healing and healing rates were determined by comparing the grid sheets. Our results found no significant difference in abrasion size between the two groups, but there was found to be a significantly improved rate of healing at follow-up in non-patched patients. This study demonstrates a significant improvement in the healing rate of traumatic CEDs in the non-patched group as compared to the patched group; therefore, the use of eye patching is not mandatory for corneal epithelial healing to occur. PMID- 9404792 TI - The use of analgesics in patients with acute abdominal pain. AB - Analgesics in patients with acute abdominal pain are often withheld for fear that they may change physical examination findings and thus may be unsafe. We conducted a randomized, prospective, placebo-controlled trial to investigate changes in physical examination following the administration of placebo, 5 mg, or 10 mg of morphine to 49 patients with acute abdominal pain. One patient was withdrawn secondary to inadequate documentation. Of the 48 patients who completed the trial, a statistically significant change in physical examination was noted in both groups receiving analgesics, but not in the placebo group. No adverse events or delays in diagnosis were attributed to the administration of analgesics. We conclude that physical examination does change after the administration of analgesics in patients with acute abdominal pain and that a larger study is needed to evaluate analgesic safety in this subpopulation of emergency department patients. PMID- 9404794 TI - Successful use of nasal BiPAP in three patients previously requiring intubation and mechanical ventilation. AB - Noninvasive mask ventilation may be used to treat patients with impending respiratory failure. In this case series, three patients with severe chronic obstructive pulmonary disease, who required mechanical ventilation in the past, were successfully treated with nasal bi-level positive airway pressure (BiPAP). All patients tolerated BiPAP well without complications. Therefore, nasal BiPAP may be considered a treatment option for patients with severe COPD who have previously required intubation and mechanical ventilation. PMID- 9404793 TI - Asymptomatic jaundice after fasting: a diagnostic dilemma. AB - An 18-year-old female with asymptomatic jaundice presented to the emergency department after fasting. She was referred to the regional medical center for evaluation and treatment. The diagnosis of Gilbert's syndrome was made by fractionation of serum unconjugated and conjugated bilirubin fraction by alkaline methanolysis, followed by thin-layer chromatography and analysis of fasting-state levels of cholyl conjugated bile acids. Methods for diagnosing this disorder are discussed. PMID- 9404795 TI - The Flexible Baton TM-12: a case report involving a new police weapon. AB - The purpose of this case report is to describe a new type of police weapon and the injuries sustained by an individual after being shot with it. The Flexible Baton is a type of ammunition that consists of a fabric bag, filled with lead shot pellets, that is shot from a shotgun. It is predicted that this bag may cause bruising, abrasions, and blunt trauma. In our patient, the fabric bag burst and the pellets penetrated the skin of the elbow, resulting in cortical violation of the distal humerus. As more guns with this ammunition have been introduced throughout the United States, we conclude that injuries from this weapon will be seen with greater frequency. PMID- 9404796 TI - Ocular examination techniques for the emergency department. AB - The purpose of this article is to provide a guide to assist the Emergency Physician in examining the eye. The evaluation of a patient with eye problems consists of a history, visual acuity, pupil examination, external examination, extra ocular movements, visual fields, and color vision. The patient is then examined at the slit lamp. After the slit lamp examination, the fundus and optic nerve is examined with a direct ophthalmoscope and intraocular pressure is measured. Special tests such as a plain film study and computed tomography (CT) scan may be obtained when indicated and, finally, referral to an ophthalmologist can be made for a dilated fundus examination, ultrasound studies of the eye and orbit, and surgical treatment. PMID- 9404797 TI - Secondary acute angle closure glaucoma: a complication of AIDS. AB - A 58-year-old man with advanced AIDS presented to the emergency department complaining of headache and decreased vision bilaterally. On evaluation, he was found to have intraocular pressures of 69 and 65 mm Hg. After topical treatment with miotics and apraclonidine, he was given intravenous acetazolamide (Diamox) and peripheral iridotomy was performed. The pressures did not improve significantly. Secondary acute angle closure glaucoma was diagnosed. Emergency physicians should consider this diagnosis when evaluating AIDS patients with visual complaints. PMID- 9404799 TI - HELLP syndrome and cholecystitis: case report and review of the literature. AB - A case of the HELLP syndrome is reported that was initially diagnosed as cholecystitis. Much overlap exists between the two diagnoses, and the emergency physician must be aware of the important differences between them. Because the HELLP syndrome and preeclampsia may occur in both the second and third trimesters, they represent serious diagnoses that must be considered when evaluating a pregnant patient with right upper quadrant abdominal pain. PMID- 9404798 TI - Pseudotoxemia: new onset psychogenic seizure in third trimester pregnancy. AB - Eclampsia, or toxemia of pregnancy, is a disorder of pregnancy characterized by seizures associated with hypertension, edema, and proteinuria. Toxemia carries significant maternal and fetal morbidity and mortality. Psychogenic seizures are defined as events that clinically resemble epileptic seizures but are not accompanied by abnormal electrical activity of the cerebral cortex. We report the case of a third trimester pregnant patient who presented with new onset convulsive activity that was associated with peripheral edema, intermittent hypertension, and proteinuria. The initial impression of the treating physicians- emergency medicine, obstetrical, and neurology--was toxemia of pregnancy. After further review and the application of numerous procedures and therapies with potential risk, the diagnosis of psychogenic seizure was made. PMID- 9404800 TI - Exotic snakebite: envenomation by an African puff adder (Bitis arietans). AB - We report an envenomation by the African puff adder (Bitis arietans), an exotic snake in the United States. The patient developed swelling and ecchymoses in the affected extremity, and cutaneous necrosis of the envenomated fingertip. There was no significant coagulopathy. He received 20 vials of specific antivenin (Schlangengift-Immunserum Behring Zentralafrika, Behringwerke, Marburg, Germany) and debridement of devitalized finger tissue. The only permanent sequelae were cutaneous scarring and permanent loss of the fingernail on the envenomated finger. Exotic snakebite is a rare presenting problem in emergency departments. The initial approach to a patient envenomated by an exotic venomous snake is discussed. Use of antivenin and supportive care are emphasized. PMID- 9404801 TI - Severe rhabdomyolysis with renal failure after intranasal cocaine use. AB - A case of acute renal failure due to rhabdomyolysis in a patient who used cocaine on a daily basis is presented. In contrast to many prior reports of renal failure occurring with cocaine-associated rhabdomyolysis, our patient did not use intravenous cocaine and did not have any evidence of trauma, seizure, hypotension, hyperthermia, hyperactivity, or coma. His creatine phosphokinase peaked at 448,000 U/liter. He was treated initially with forced diuresis and i.v. furosemide, but he became oliguric, developed pulmonary edema, and required hemodialysis. He recovered fully after 3 weeks of dialysis. The literature is reviewed in an attempt to delineate a rational approach to evaluating cocaine users at risk for rhabdomyolysis. PMID- 9404802 TI - Beyond the twelve-lead electrocardiogram: diagnostic tests in the evaluation for suspected acute myocardial infarction in the emergency department, part I. AB - On a daily basis the emergency physician is faced with the difficult task of determining whether or not a patient with acute chest pain is sustaining an acute myocardial infarction. In most cases, this is not a straightforward decision. Although observation units are being used more often for chest pain evaluations, many emergency physicians currently admit such patients to an intensive care setting. Because fewer than one-third of emergency department chest pain patients actually suffer an acute myocardial infarction, expensive resources are, in retrospect, used unnecessarily. Conversely, patients who are infarcting, and are inadvertently discharged home from the emergency department, have a worse prognosis than those admitted. This two-part series reviews the newer modalities available that may help the emergency physician arrive at a more accurate diagnosis. The current article, Part I, examines the use of myocardial imaging, computer assisted diagnostic protocols, and newer uses of the electrocardiogram. Part II reviews the use of biochemical assays of cardiac proteins and the Chest Pain Observation Unit. PMID- 9404803 TI - Strangulated traumatic diaphragmatic hernia simulating a subphrenic abscess. AB - Traumatic diaphragmatic hernias can be difficult to diagnose because of their varied clinical and radiologic signs and because patients may not present with symptoms for months to years following the injury. We report a case of a delayed presentation of a traumatic diaphragmatic rupture through which a portion of the stomach herniated and simulated a large subphrenic abscess. PMID- 9404804 TI - Another false alarm? apnea monitor activation in a Neonatal Intensive Care Unit graduate. AB - Neonatal emergencies have become more common as increasingly sophisticated Neonatal Intensive Care Units graduate lower birth-weight babies born at younger gestational ages. These patients present a number of challenges to emergency physicians. They are often discharged with apnea monitors, which generate a high number of false alarms. Neonatal Intensive Care Unit graduates, however, are predisposed to a number of conditions that can result in true episodes of apnea. We present such a case and will discuss the unusual underlying cause of apnea, the utility of apnea monitors, and the need for emergency physicians to be prepared to evaluate and treat these potentially complicated patients. PMID- 9404805 TI - Acute thoracic aortic dissection: the basics. AB - With an increasing incidence, aortic dissection is the most common acute illness of the aorta. In the setting of chronic hypertension, with or without other risk factors for aortic dissection, this diagnosis should be considered a diagnostic possibility in patients presenting to the emergency department with acute chest or back pain. Left untreated, about 75% of patients with dissections involving the ascending aorta die within 2 weeks of an acute episode. But with successful initial therapy, the 5-year survival rate increases to 75%. Hence, timely recognition of this disease entity coupled with urgent and appropriate management is the key to a successful outcome in a majority of the patients. This article reviews acute thoracic aortic dissection, including ED diagnosis and management. PMID- 9404806 TI - The erythrocyte sedimentation rate. AB - The erythrocyte sedimentation rate has been used for over 50 years for everything from predicting disease severity to assessing general sickness index. Its perceived utility has been based on medical myths and its use too often based only on a consultant's demand or a shotgun approach to diagnosis. This article focuses on examining the specific utility of the erythrocyte sedimentation rate in the emergency department as a tool for predicting both disease likelihood and severity. PMID- 9404807 TI - Acute flank pain. PMID- 9404808 TI - Unilateral proptosis. PMID- 9404809 TI - Broken wings. PMID- 9404810 TI - Broken wings, mandalas, impaired physicians, and medical examining boards. PMID- 9404811 TI - Emergency department triage: a program assessment using the tools of continuous quality improvement. AB - An assessment was undertaken in the emergency department of a busy tertiary care center to illustrate the role of continuous quality improvement in the evaluation of an emergency triage program that utilizes the emergency medical attendant to provide triage. An evaluation team interviewed triage staff, charge nurses, internal customers, risk management, and the patient representative. A detailed review of staff job descriptions, organization charts, orientation manual, and physical facilities was conducted. A chart audit was completed on 100 triage notes. Direct observation was undertaken on nine occasions. An evaluation of the data gathered was performed using the tools of continuous quality improvement, and resulted in specific recommendations being made to improve the process of care. It was concluded that emergency medical attendants function very well in an emergency medicine triage system and the tools of continuous quality improvement can be applied to a clinical service to improve the quality of care. PMID- 9404812 TI - Cerebral penetration injury leads to H2O2 generation in microdialysis samples. AB - Delayed tissue damage is proposed to be caused by reactive oxygen species. We investigated the effects of microdialysis probe penetration into rat piriform cortex on hydrogen peroxide (H2O2) in brain extracellular fluid (ECF). H2O2 decreased immediately after probe insertion into the brain, but increased over 300% in samples within minutes after collection. We assessed H2O2 changes in vitro in microdialysis perfusion media containing various ascorbic acid concentrations and confirmed ascorbic acid is a source of H2O2. We conclude that decreased H2O2 concentrations in perfusion media as it passes through the brain reflect an extracellular antioxidant effect, whereas the increase in H2O2 with time after sample collection indicates that H2O2 generating substances are present in ECF. Thus, the potential for producing reactive oxygen species in brain ECF exists following penetration injury, especially if transition metals are released into the neuronal microenvironment. PMID- 9404813 TI - Effects of pinealectomy on SCN electrical firing rhythm in Djungarian hamsters. AB - Djungarian hamsters (Phodopus sungorus) from a long day photoperiod (16:8 h light dark cycle) were either pinealectomized (PINX; n = 7) or sham-pinealectomized (SHAM; n = 6). One week after surgery coronal brain slices (500 microm), containing the suprachiasmatic nucleus (SCN), were prepared. The firing-rate rhythm of SCN neurons was recorded over at least a 22 h period. The amplitude of the firing rhythm for SHAM slices (3.42 +/- 0.26 Hz) was not significantly different from PINX slices (3.45 +/- 0.29 Hz). Maxima and minima of the firing rhythms also could not be distinguished statistically between PINX and SHAM groups. However, two-way ANOVA of 2 h firing-rate averages indicated a statistically significant effect of pinealectomy on the firing rhythm (P < 0.01). The results show that pinealectomy one week prior to brain slice preparation only had a minor effect on the in vitro neuronal firing rhythm in the SCN. In this study, SCN firing-rate rhythm in the Djungarian hamster is largely independent of endogenous melatonin secretion. PMID- 9404814 TI - Diet can modify autotomy behavior in rats following peripheral neurectomy. AB - Partial sciatic nerve ligation in rats (PSL) produces neuropathic pain disorders [Seltzer, Z., Dubner, R. and Shir, Y., Pain, 43 ( 1990) 205-18 (corrected) ]. Recently we reported that diet markedly affected the levels of these disorders. Here we questioned whether diet also affects neuropathic pain-related behavior in another model, produced by total denervation of the hindpaw following peripheral sciatic and saphenous neurectomy. Sabra rats and HA line rats were fed for 2-3 weeks preoperatively and up to 58 days postoperatively (PO) with one of five different diet formulas. We found that the autotomy behavior differed significantly between the diet groups. Surprisingly, in some diets the effects on autotomy and PSL models were different and even contrasting. Modulation of diet in humans may emerge as a novel therapy of neuropathic pain. PMID- 9404815 TI - The nitric oxide synthase expression of rat cortical and hippocampal neurons changes after early lead exposure. AB - The effect of lead exposure in nitric oxide synthase containing neurons (nNOS) within rat cortex and hippocampus was studied. Lead administration (1 g% lead acetate in drinking water) was commenced prior to mating and continued until 30 postnatal (PN) days. Immunohistochemical studies using antibody to nNOS showed, after lead treatment at PN21-PN30, a reduction in neuronal size and optical density (OD) of nNOS+ cells. In both regions, non-pyramidal immunoreactive neurons exhibited smaller soma size and less developed dendrites. A significant difference in cell areas and OD of lead exposed versus control rats and no variation in the number of nNOS+ neurons was seen. Morphological modifications after early lead exposure, induced nNOS reduction in NOS expressing neurons thereby interfering in NO synthesis. PMID- 9404816 TI - Effects of potassium channel blockers on the acetylcholine-induced currents in dissociated outer hair cells of guinea pig cochlea. AB - Much physiological evidence is available to show that acetylcholine (ACh) hyperpolarizes the outer hair cells (OHCs) of guinea pig cochlea and induces Ca2+ activated K+ currents. In this study, using the nystatin perforated patch-clamp technique, we investigated the effects of various K+ channel blockers on the ACh induced currents (I[ACh]) in dissociated OHCs of guinea pig cochlea. The I(ACh) were inhibited by apamin in a concentration-dependent manner. The half-maximal inhibitory concentration for apamin on the ACh-induced response was 1.59 x 10(-9) M. Charybdotoxin and iberiotoxin had no inhibitory effect on the I(ACh) The inhibitory potency of the various K+ channel blockers on the I(ACh) was as follows: apamin >> quinine approximately quinidine approximately d-tubocurarine > tetraethylammonium chloride > 4-aminopyridine approximately Ba2+ > Cs2+. It is proposed that the Ca2+-activated K+ channels of mammalian cochlear OHCs should be classified as small conductance Ca2+-activated K+ (SK) channels according to their pharmacological properties. PMID- 9404817 TI - Localization of D1 and D2 dopamine receptors in the rat mesencephalic trigeminal nucleus by immunocytochemistry and in situ hybridization. AB - The rat mesencephalic trigeminal nucleus (MTN) receives a dopaminergic innervation. In order to identify and localize dopaminoceptive cells within this nucleus, expression of D1 and D2 dopamine (DA) receptors was examined by immunocytochemistry with subtype-specific antibodies and in situ hybridization with digoxigenin-labeled cRNA probes. Immunocytochemical labeling was restricted to neuronal perikarya and proximal processes whereas the hybridization signal was confined to MTN cell bodies. Cells immunopositive for D1 were located throughout the entire MTN whereas cells labeled with D2 antibodies were concentrated in its caudal portion. D1 receptor message was found in relatively low levels. In contrast, high levels of mRNA for D2 were seen in MTN neurons. The distribution of DA receptor mRNA-containing cells were very similar to those of DA receptor immunoreactivity. Neurons expressing the D1 receptor gene were localized in both rostral and caudal portions, which receive inputs from masticatory muscle spindles and from spindles and periodontal ligament receptors, respectively. D2 receptors were limited to ventrocaudally located cells. These results suggest that processing of proprioceptive information in the MTN is controlled by the DAergic input through different postsynaptic mechanisms. PMID- 9404818 TI - Etomidate inhibits [3H]noradrenaline release from SH-SY5Y human neuroblastoma cells. AB - We have examined the effects of the intravenous anaesthetic induction agent etomidate on K+ and carbachol evoked [3H]noradrenaline ([3H]NA) release and the associated increase in [Ca2+]i in SH-SY5Y human neuroblastoma cells in a attempt to study potential anaesthetic target site(s). Preincubation with etomidate produced a dose-dependent inhibition of both K+ and carbachol evoked [3H]NA release with estimated IC50 values of 88 and 69 microM, respectively. Only K+ stimulated increase in [Ca2+]i was inhibited by etomidate preincubation with an IC50 of 146 microM. Acute addition of etomidate after K+ challenge also inhibited the increase in [Ca2+]i with an IC50 of 99 microM. In addition etomidate displaced the binding of [3H]PN200-110 to L-type voltage sensitive Ca2+ channels with a Ki of 48 microM. As K+ but not carbachol evoked [3H]NA release is extracellular Ca2+ dependent and was inhibited by etomidate these data coupled with the PN200-110 displacement studies suggest that etomidate may interact with L-type voltage sensitive Ca2+ channels. The inhibition of carbachol evoked release without affecting the associated increase in [Ca2+]i suggests that etomidate may exert additional effects at either the muscarinic receptor or the secretory machinery in these cells. PMID- 9404820 TI - Changes of expression levels of choline acetyltransferase and vesicular acetylcholine transporter mRNAs after transection of the hypoglossal nerve in adult rats. AB - Acetylcholine, synthesized in the cytoplasm of cholinergic neurons by choline acetyltransferase (ChAT), is packaged in synaptic vesicles by vesicular acetylcholine transporter (VAChT). The entire VAChT gene has been reported to be located within the first intron of the ChAT gene. In order to examine whether or not ChAT and VAChT transcription may be coordinately regulated, the levels of ChAT and VAChT mRNAs in hypoglossal neurons were analyzed by in situ hybridization following transection of the hypoglossal nerve in adult rats. After unilateral transection, the levels of expression of ChAT and VAChT mRNAs were dramatically reduced in the ipsilateral hypoglossal nucleus 1 week after the surgery. However the expression of both mRNAs gradually recovered thereafter. These results suggest that the transcription of the two cholinergic genes is tightly linked in motor neurons. PMID- 9404819 TI - N-acetylaspartylglutamate (NAAG) protects against rat striatal quinolinic acid lesions in vivo. AB - We examined the effects of N-acetylaspartylglutamate (NAAG), an endogenous peptide thought to be involved in neurotransmission and neuromodulation, on striatal quinolinate lesions, a rodent model of Huntington's disease. We found that NAAG (500 and 1000 nmol) co-injected with quinolinic acid significantly reduced lesion volumes (by 50% and 65%, respectively). A 1000 nmol dose of the non-hydrolyzable analogue, beta-NAAG, also reduced quinolinic acid lesion volumes by 78.4%, indicating that the protection observed was not secondary to cleavage of NAAG into N-acetyl-aspartate (NAA) and glutamate. Likewise, co-injection of both NAA and glutamate (1000 nmol each) with quinolinic acid did not significantly alter the size of lesions. NAAG's protective effect may be mediated through actions on N-methyl-D-aspartate receptors or metabotropic glutamate receptors. PMID- 9404821 TI - Pontine neurons which relay projections from the superior colliculus to the posterior vermis of the cerebellum in the cat: distribution and visual properties. AB - Extracellular unit recording revealed that the dorsolateral pontine nucleus (DLPN) and nucleus reticularis tegmenti pontis (NRTP) of the cat constituted pontine relays for transmission of visual information from the superior colliculus (SC) to the posterior vermis (lobules VI and VII) of the cerebellum. The relay neurons in DLPN/NRTP responded to moving visual stimuli with large receptive fields (23-75 degrees ) which occupied mainly the contralateral hemifield including the fovea. These neurons were usually more responsive to large-sized stimuli than to discrete-spot stimuli, had direction selectivity, and showed preference to visual motion at a relatively high speed. No clear differences in visual properties were observed between DLPN and NRTP. After a local injection of lidocaine into SC, the visual responses in DLPN/NRTP transiently disappeared, indicating that DLPN/NRTP received the visual inputs through SC. PMID- 9404822 TI - Reduced vagal sensory innervation of the small intestinal myenteric plexus following capsaicin treatment of adult rats. AB - To determine whether capsaicin treatment damages small intestinal vagal sensory nerve endings, we made intra-nodose injections of wheat-germ agglutinin horseradish peroxidase conjugate (WGA-HRP) to label peripheral and central vagal endings in control and capsaicin-treated rats. Labeled intraganglionic laminar endings (IGLEs), characteristic of vagal sensory endings of the myenteric plexus, were counted. In controls IGLEs were numerous in the duodenum, less numerous in the jejunum and scarce in the ileum. In capsaicin-treated rats, IGLEs were significantly diminished in all areas of the small intestine. Capsaicin also reduced WGA-HRP activity in the medial and commissural nucleus of the solitary tract. Systemic capsaicin produces a long-lasting loss of vagal IGLEs in the small intestinal myenteric plexus. Such loss is consistent with capsaicin-induced impairment of intestinal reflexes and controls of food intake. PMID- 9404823 TI - Modulation of P-glycoprotein activity by glial factors and retinoic acid in an immortalized rat brain microvessel endothelial cell line. AB - P-glycoprotein (P-gp), a product of the multidrug-resistant (mdr) genes, is expressed in the endothelial cells of the blood-brain barrier (BBB). Effects of glial factors and retinoic acid (RA) on P-gp activity and level were investigated in the immortalized rat brain endothelial cell line RBE4, which expressed immunodetectable P-gp associated with a decrease in accumulation of the P-gp substrates, vinblastine and colchicine. When RBE4 cells were cultured either in the presence of C6-conditioned medium or on C6- or astrocyte-extracellular matrix, intracellular vinblastine and colchicine concentrations were decreased. When the cells were treated with RA, increases in P-gp activities were correlated with increases in P-gp levels. Effects of simultaneous treatments with glial factors and RA were studied in RBE4 cells cultured on astrocyte-extracellular matrix and were shown to be additive on P-gp activity and level. RBE4 cells may serve as a useful in vitro model for basic research on P-gp regulation at the level of the BBB. PMID- 9404824 TI - Cortical [3H]ketanserin binding and 5-HT2A receptor-mediated inositol phosphate production in the spontaneously hypertensive rat and Lewis rat strains. AB - The spontaneously hypertensive rat (SHR) and Lewis rat strains differ in the elevated plus-maze of anxiety, the former and the latter strain displaying low and high anxiety, respectively. A recent study has shown that serotonin (5 hydroxytryptamine, 5-HT)2A receptor-mediated head shakes, but not [3H]ketanserin binding at these receptors, are less in Lewis rats, compared with SHRs. Herein, we have analysed the hypothesis of a difference in 5-HT2A receptor-effector coupling between these two strains. Confirming our previous results, the Bmax and KD values for the specific [3H]ketanserin binding at cortical 5-HT2A receptors were respectively identical in both strains. The accumulation of total inositol phosphates by the 5-HT2A,2B,2C receptor agonist 1-(2,5-dimethoxy-4-iodophenyl)-2 aminopropane (DOI; 0.01-100 microM) was concentration- and strain (Lewis > SHR) dependent. Preincubation with 0.01 and 0.1 microM of the 5-HT2A receptor antagonist SR 46349B, respectively, decreased and prevented DOI-elicited inositol phosphate production in both strains. The aforementioned genetic differences in 5 HT2A receptor-mediated head shakes may thus lie at some point distal from the 5 HT2A receptor. PMID- 9404825 TI - A role of periaqueductal grey NMDA receptors in mediating formalin-induced pain in the rat. AB - In the present study we examined the role of periaqueductal grey (PAG) N-methyl-D aspartate (NMDA) receptors in the perception of tonic and phasic pain. Under sodium pentobarbital anesthesia rats were implanted unilaterally with a guide cannula aimed at the PAG. Following a 7-14 day recovery period rats received an infusion of the NMDA antagonist, 2-amino-5-phosponopentanoic acid (AP5), or saline into the PAG. Five minutes after the infusion of AP5 rats were tested for analgesia in the formalin test, or in the hotplate test. AP5 injections into the PAG reduced pain in the formalin test, but not the hotplate test. These data show that NMDA receptors within the PAG are involved in the perception of tonic, inescapable pain as measured in the formalin test, but not phasic, escapable pain as measured in the hotplate test. PMID- 9404826 TI - Characterization of growth/differentiation factor 5 (GDF-5) as a neurotrophic factor for cultured neurons from chicken dorsal root ganglia. AB - Growth/differentiation factor-5 (GDF-5), a morphogenetic protein, has previously been shown to act as a neurotrophic factor for midbrain dopaminergic neurons. To further elucidate the neurotrophic potential of GDF-5, serum free cultures of dorsal root ganglionic (DRG) neurons from developing chick embryos were treated with GDF-5 with or without the simultaneous addition of other trophic factors. Our results show that GDF-5 has a minor promoting effect on its own, but it can enhance the survival promoting effect of neurotrophin-3 (NT-3) and nerve growth factor (NGF) on cultured DRG neurons. Our finding fits well into the concept that neurotrophic factors may act synergistically in ensuring survival of different neuronal populations. The capacity of GDF-5 to reduce the requirement of a subpopulation of sensory neurons for NT-3 may have implications for the treatment of peripheral neuropathies. PMID- 9404827 TI - The use of multisite high resolution arterial imaging to assess arteriosclerosis. AB - This article presents a model for the use of multisite, sonographic imaging to assess arteriosclerosis. The arteries of 100 randomly selected patients were scanned in three anatomical areas (carotid, femoral-popliteal, aorta-iliac) in conjunction with measurement of selected risk factors (smoking, cholesterol, triglycerides, high density lipoproteins, antioxidant levels). Arteries were interrogated for intimal wall hyperplasia, plaque, and ulceration. Multisite scanning (24 sites) detected the presence of pathology in all sites surveyed. The model was developed at the Cardiovascular Wellness Center in Westbury, NY. PMID- 9404828 TI - Heparin-induced extracorporeal fibrinogen/LDL precipitation (HELP): a promising regimen for the treatment of vascular diseases. AB - Current management of atherosclerotic diseases consists primarily of medical therapy designed to increase oxygen supply to the heart, the brain, retinal vessels, or lower limbs. The development of these diseases is based on atherosclerotic changes induced by risk factors such as elevated levels of fibrinogen and lipoproteins. These risk factors are related to a dramatic deterioration of the hemorrheologic pattern, which reduces perfusion. Consequently, attempts are now being made to treat ischemia via hemorrheological intervention. A new treatment modality utilizing the heparin-induced extracorporeal low-density lipoprotein (LDL) precipitation (HELP), offers the possibility of obtaining therapeutic success not only in cases of severe hypercholesterolemia but also in the field of hemorheology. With HELP a safe and rapid reduction of fibrinogen and lipid fractions has become feasible, thus providing acute improvements of red cell aggregation, of the filterability of blood cells, of whole-blood and plasma viscosity, and thereby of microcirculation. Because cerebrovascular diseases are known to be related to disturbances of the hemorrheological situation, the HELP system is used in the Department of Electrobiology of Graz for the treatment of acute stroke, cerebral multi-infarct disease, and occlusions of retinal arteries. PMID- 9404829 TI - Raynaud's phenomenon in vibration syndrome: the impact of cold feet on skin temperature and vasomotion of the hand after immersion in cold water. AB - Patients with vibration syndrome, suffering from Raynaud's phenomenon, are sensitive to cold. Rewarming time, after local cooling, is delayed. The present study evaluated whether rewarming of the hand after cooling is influenced by the temperature of the feet. In five Japanese patients (former forest workers) with vibration syndrome, suffering from Raynaud's phenomenon, and in five healthy controls, temperature changes of the hand after cooling were registered under the two test situations (on different days) with the feet immersed in water of 35 degrees C or 20 degrees C, respectively. In both patients and controls (in both groups, in four of five cases) rewarming of the hand after cooling was faster when the feet were immersed in cold water, compared with when the feet were immersed in warm water. In this test situation, the systemic thermoregulative counterreaction appears to be more important for rewarming of the hand after cooling than a possible synchronous passive reaction accompanying warming of the feet. A deliberate training of the systemic counterreaction may prove beneficial for patients with Raynaud's phenomenon. PMID- 9404830 TI - Errors of computer electrocardiography. AB - A review of 6938 computer-generated electrocardiogram interpretations was undertaken by a single cardiologist. The computer-assisted program correctly interpreted 51.6% of the tracings. One or more corrections were necessary in 48.4%. The greatest number of incorrect interpretations involved repolarization (T wave) abnormalities, followed by ST segment changes. It is mandatory that computer-generated ECG interpretations be reviewed by an experienced electrocardiographer. Reliance on nonreviewed ECG interpretations may result in incorrect diagnoses that could lead to inappropriate management decisions. PMID- 9404831 TI - Acute electrophysiological effects of dipyridamole on sinus node function in patients with sick sinus syndrome. AB - One of the most widely used tests for evaluation of sinus node function is sinus node recovery time (SNRT), which requires right heart catheterization. On the other hand SNRT has high specificity but only moderate sensitivity in the diagnosis of sick sinus syndrome (SSS). The authors studied acute electrophysiologic effects of dipyridamole (0.40 mg/kg IV) in 16 patients with clinical SSS. All of them had normal SNRT and had undergone permanent DDD pacemaker implantation. By the aid of temporary pacing inhibition, the authors noninvasively measured the corrected sinus node recovery time (SNRTc) and sinus cycle length (SCL) before and after dipyridamole administration. SCL was slightly decreased from a mean basal value of 1025 +/-323 to 913+/-213 msec after dipyridamole administration (mean -10%), but this was not statistically significant. SNRTc was increased from a mean basal value of 344+/-91 to 606+/-156 msec after dipyridamole administration (+76% P< or =0.004). These results suggest that dipyridamole must be used cautiously in patients with SSS. Intravenous dipyridamole may be a useful test to assess sinus node function. SNRT measurement after intravenous dipyridamole may increase sensitivity of this test in patients with suspected SSS and normal SNRT. PMID- 9404832 TI - Evaluation of normal hemodynamic profile of CarboMedics prosthetic valves by Doppler echocardiography. AB - The authors investigated 163 CarboMedics bileaflet prosthetic valves--81 mitral prostheses (MP), and 82 aortic prostheses (AP)--to determine acceptable pressure gradients across normally functioning prostheses and effective mitral valve orifice (MVO) area by Doppler echocardiography. In MP, the mean gradient was 3.6+/-1.7 mm Hg, peak transmitral gradient was 8.7+/-3.7 mm Hg, and mean effective valve area was 2.3+/-0.7 cm2. There was a significant overlap in mean and peak transaortic gradients even with valves of the same size. In AP, the mean gradient was 14.7+/-5.1 mm Hg and peak pressure gradient was 26.1+/-8.2 mm Hg. They observed a weak inverse correlation between valve size and gradients in AP. Mean and peak pressure gradients tended to be higher with smaller valve sizes, but differences were statistically significant (P < 0.5) only when they compared the smallest vs the largest valves. Trivial to mild regurgitation was detected in 28.4% of MP and 54.8% of AP. From the data they conclude that CarboMedics valves offer relatively little resistance to forward flow, both in the mitral and aortic positions, and their hemodynamic profile is comparable to that of the St. Jude bileaflet valves described in published literature. PMID- 9404833 TI - Aortic dissection in a patient receiving chemotherapy for Hodgkin's disease--a case report. AB - Cancer chemotherapy is associated with a wide range of vascular toxicities, which may be related to endothelial cell damage by these agents. The authors describe a patient with Hodgkin's disease who developed an atypical aortic dissection while receiving MOPP/ABV chemotherapy (nitrogen mustard, vincristine, procarbazine, prednisone, doxorubicin, bleomycin, and vinblastine). They would place aortic dissection on the list of potential vascular complications associated with antineoplastic agents. PMID- 9404835 TI - Giant coronary artery pseudoaneurysm causing pulmonary artery obstruction: a rare complication of coronary bypass surgery--a case report. AB - The authors report the diagnosis and successful management of a 57-year-old man with right ventricular outflow tract obstruction from a large pseudoaneurysm of the left anterior descending coronary artery 5 years after he had undergone redo coronary artery bypass grafting. PMID- 9404834 TI - Three-dimensional spiral computed tomography angiography as an alternative imaging modality for a silent dissecting aortic aneurysm--a case report. AB - The authors report a patient with silent dissecting aortic aneurysm in whom three dimensional spiral computed tomography (CT) angiography (3D-CTA) provided important imaging data. Images obtained by 3D-CTA were compared with the results of both conventional angiography and CT. They conclude that 3D-CTA was a powerful diagnostic modality for this patient, in addition to conventional CT and angiography. PMID- 9404836 TI - Pulsus alternans in diastolic left ventricular dysfunction--a case report. AB - Pulsus alternans is usually found in patients with reduced systolic ventricular function. We describe a patient with recurrent pulmonary edema, hypertension, bilateral renal artery stenosis, but with normal systolic function. Pulsus alternans was demonstrated in both pulmonary artery, right ventricle, and left ventricle pressures. After successful renal artery revascularization, the pulsus alternans disappeared. This case illustrates that pulsus alternans can be present with diastolic dysfunction of the left ventricle in the absence of systolic dysfunction. PMID- 9404837 TI - Some factors affecting the prevalence of Trypanosoma evansi in camels in Mauritania. AB - A study was conducted on the epidemiology of camel trypanosomosis in Mauritania using 2073 camels of various ages in five regions (Trarza, Gorgol, Adrar, Hodh E1 Chargui, Nouakchott). The prevalence was determined through blood smear and serological tests: card agglutination test for trypanosomiasis (CATT) and immuno fluorescence antibody test (IFAT). The prevalence of the disease was 1.3% using blood smear examinations, 16.2% with CATT and 25.2% with IFAT. The following variations were observed: (1) Camels in Trarza had the highest prevalence; (2) Intraregion was a significant factor; (3) Animals that migrated to the south were more commonly infected than those in the north; and (4) Animals in the 5- to 10 yr age group had the highest prevalence. The study indicated that camel trypanosomosis was widespread in Mauritania, especially in the wooded areas near waterways in the south. PMID- 9404838 TI - Class-specific antibody responses in cattle following experimental challenge with sporocysts or merozoites of Sarcocystis cruzi. AB - An ELISA using antigen produced from merozoites of Sarcocystis cruzi was developed to monitor specific IgM and IgG antibody, following challenge of cattle with either merozoites or sporocysts of S. cruzi. This assay was compared with an ELISA using antigen produced from the cystozoite stage of the parasite. Both ELISAs were able to detect significant increases in levels of circulating IgM and IgG antibodies against Sarcocystis in all challenged cows; however, the magnitude of the titres was greater in the ELISA which used the antigen derived from the merozoites. This immunoassay also detected increases in the levels of IgG earlier than did the assay using antigen derived from cystozoites of S. cruzi. Since this rise coincided with the presence of clinical signs, and was persistent for several weeks, the IgG-ELISA using antigen derived from merozoites appears to be suitable for the diagnosis of acute sarcocystiosis in cattle. Furthermore, since significant increases in the levels of circulating IgM and IgG antibodies against Sarcocystis were detected in the cows infected with merozoites of S. cruzi, it is evident that merozoites of S. cruzi cultured in vitro maintain their capability to replicate in the natural intermediate host. PMID- 9404839 TI - Clinical observations, pathology, bioassay in mice and serological response at slaughter in pigs experimentally infected with Toxoplasma gondii. AB - Experimental infections of a total of 47 pigs with tachyzoites of the Toxoplasma gondii RH-strain, tissue cysts of the SSI-119 and R92 strains as well as oocysts of the SSI-119 strain were performed to determine the sensitivity of an indirect IgG-ELISA, using tachyzoite lysate of the RH-strain as antigen. The infections led to a dose dependent moderate clinical affection (inappetence, fever and poor general condition). Pigs infected with 10000 oocysts or with 1/2 mouse brain containing tissue cysts of the SSI-119 strain showed a significant decrease in weight gain compared to uninoculated pigs during the first 2 weeks p.i., followed, however, by compensatory growth during the next 6 weeks. At slaughter 3 to 4 months after inoculation 39/41 (95.1%) of pigs positive by bioassay in mice were seropositive in ELISA. Tissue cysts were not demonstrable by immunohistochemistry. ELISA OD-values obtained by analysis of meat juice from heart muscle and tongue (diluted 1:40) correlated strongly with OD-values by analysis of serum (diluted 1:400) (r heart juice = 0.942; r tongue juice = 0.915). Thus, meat juice samples were shown to provide a suitable alternative to serum for serological detection of Toxoplasma infection in pigs. PMID- 9404840 TI - The presence of Cryptosporidium oocysts in stools of clinically diarrhoeic and normal nonhuman primates in Kenya. AB - A total of 114 nonhuman primates comprising 51 vervet monkeys (Cercopithecus aethiops) and 63 olive baboons (Papio anubis) were examined for Cryptosporidium oocysts using the modified Kinyoun's acid-fast staining technique. About 51.7% (59/114) of all the specimens examined, representing 78.4% (40/51) of the vervet monkeys and 30.1% (19/63) of the olive baboons were positive. Bright red, refractile Cryptosporidium oocysts were observed in the stained faecal smears against a blue background. Up to 4/6 (66.7%) of the diarrhoeic vervets and 2/3 (66.7%) baboons, respectively, were positive while the rest were negative. To the best of our knowledge, this report is the first on cryptosporidiosis in old world nonhuman primates in Kenya and probably the first report of the infection in olive baboons. Given the high frequency of oocysts in diarrhoeal specimens, the parasite may have been associated with clinical diarrhoea in the sampled animals. Cryptosporidium, which has been reported in humans in Kenya, is also suspected to occur in livestock. Its isolation from clinically ill, normal colony-borne and newly caught feral nonhuman primates has significant implications for both public health and animal agriculture in Kenya. PMID- 9404841 TI - The potential of nematophagous fungi to control the free-living stages of nematode parasites of sheep: comparison between Australian isolates of Arthrobotrys spp. and Duddingtonia flagrans. AB - Nine isolates of Duddingtonia flagrans and eight isolates of Arthrobotrys spp. which originated from a field survey for the presence of nematophagous fungi in fresh dung of livestock in Australia were used in this study. Comparisons were made between the ability of the different isolates to survive gut passage and subsequently reduce infective larval numbers in sheep faeces. Fungal spores (conidia and/or chlamydospores) were administered orally to sheep in doses ranging from 1 X 10(5) to 4.5 X 10(6) spores. There was no apparent consistent survival of Arthrobotrys spp., whereas D. flagrans showed excellent survival capacity which resulted in profound reductions in Trichostrongylus colubriformis larval numbers in culture. This provides clear evidence that D. flagrans is an ideal candidate as a potential biological control agent for nematode parasites of sheep. PMID- 9404842 TI - Quality control in generic anthelmintics: is it adequate? AB - We became increasingly concerned about indications of possible substandard efficacy of some generic anthelmintics, particularly after P.C. van Schalkwyk (personal communication, 1990) had found some batches of imported generic products obtained from international brokers to be poorly active, despite apparently normal physical characteristics. Therefore, considering the serious consequences this would have for sheep farming, it was decided to test the efficacy of some of the generic rafoxanide products available on the South African market. One of the three commercial formulations (of highly reputable companies) tested against a known susceptible strain of Haemonchus contortus in sheep was markedly substandard, with an arithmetic mean efficacy of 66.2% (Class B, Reinecke, 1973), compared to Class A efficacy of the other two, which also differed significantly from one another (Mann-Whitney; P = 0.01). Larger differences were found between the three products against a natural infection with a partially resistant strain of H. contortus than against the susceptible strain, with corresponding arithmetic mean efficacies of 28.7% (Class X, or ineffective), 71.3% (Class B) and 87.7% (also Class B). It is concluded that the most likely reason for the observed differences is that international brokers do not disclose the sources of supply of different batches of active ingredient (with the result that the companies buying anthelmintics from them have no way of telling when a source of supply is changed); that the efficacy of such batches differs; and that efficacy testing of individual batches in some cases is inadequate. It is suggested that registering authorities should consider simplified efficacy testing of each new batch of active ingredient before it may be marketed. PMID- 9404843 TI - Clinical trial of moxidectin oral gel in horses. AB - A clinical trial carried out over 98 days was done to evaluate treatment of horses with moxidectin gel for efficacy as measured by (1) reduction in the production of parasite ova post treatment, (2) a comparison of the posttreatment parasite egg count suppression of moxidectin to ivermectin, and (3) assessment of the field safety, animal acceptance of the moxidectin formulation, and the utility of the moxidectin delivery device. One hundred and fifty Standardbred horses with naturally acquired parasite infections were used in the study. Moxidectin had more prolonged and greater suppressive influence than did ivermectin on reappearance and magnitude of strongyle egg counts post treatment. Differences were not observed between the capability of ivermectin or moxidectin to reduce and suppress low Parascaris equorum egg counts. Adverse reactions to treatments were not observed, and the utility of the moxidectin delivery syringe and animal acceptance of moxidectin treatment were satisfactory. PMID- 9404844 TI - Comparative curative and preventive efficacies of ivermectin and closantel on Oestrus ovis (Linne 1758) in naturally infected sheep. AB - A field trial was undertaken to assess the efficacy of each of two formulations of ivermectin and of closantel in prevention and treatment of Oestrus ovis in a naturally infected flock grazing on the foothills of the Pyrenees mountains, in south-western France. Within the flock, 875 sheep were randomly divided into four groups, and treated twice during the fly season, with an interval of 60 days between treatments. Group 1 sheep were treated with albendazole (ABZ) at a dose rate of 3.8 mg/kg to maintain control of trichostrongylid parasites without affecting O. ovis; Group 2 received closantel at a dose rate of 10 mg/kg because of its known persistent activity against O. Ovis; Groups 3 and 4 received ivermectin at a dose rate of 200 mcg/kg bodyweight by subcutaneous injection (Isc) and orally (Io), respectively. All sheep were managed as a single group throughout the study. In order to assess the prophylactic effect of each product, immediately prior to the scheduled second treatment on Day 60 (D60), five sheep from each group were chosen at random and necropsied. Similarly, to assess the therapeutic effect, another five sheep from each group were selected on D70 and necropsied for parasite counts. During the 120 days of the trial, a significant number of animals from each group were regularly individually examined to assess their clinical status with regard to O. Ovis infection. Clinical signs of infection had significantly declined in Groups 2, 3 and 4 by 10 days after treatment reaching their lowest level at D30. In the control group during this period, clinical signs increased. Ten days after the second treatment, (D70), there was also evidence of a significant response to treatment. Finally the between-treatment differences in clinical scores of the closantel and ivermectin groups were small, although scores in Group 1 sheep was suggestive of a higher challenge in the second half of the study. On the basis of the postmortem counts and arithmetic means, prophylactic efficacies for the treatments relative to ABZ treated group, were 97.7, 62.5 and 0%, for the closantel, Isc and Io groups respectively. Therapeutic efficacies for the closantel, Isc and Io were 100, 100 and 98% respectively. PMID- 9404845 TI - Construction and initial analysis of a representative lambda ZAPII expression library of the intracellular rickettsia Cowdria ruminantium: cloning of map1 and three other Cowdria genes. AB - The causative agent of heartwater, the rickettsia Cowdria ruminantium, is very poorly understood at the molecular level owing to a profound lack of suitable tools. We have developed an immunoaffinity chromatographic method to purify C. ruminantium from host cell components and the purified rickettsial cells have been used to prepare substantially pure Cowdria DNA. This DNA has been used to construct what we believe to be the first fully representative C. ruminantium expression library. A clone containing the complete Cowdria map1 gene has been isolated and sequenced. This gene has been expressed in E. coli cells from the native Cowdria promoter, suggesting that the mechanisms for gene transcription and translation are similar between these two organisms. Parts of three other Cowdria genes have also been isolated and sequenced. PMID- 9404846 TI - Epidemiological investigation of trichinellosis in Switzerland. AB - Domestic pigs in Switzerland have been considered Trichinella-free for decades, despite the occurrence of Trichinella in the wildlife cycle. In order to reevaluate the present epidemiological situation, tissue samples from 11226 domestic pigs, 356 wild boars and 452 foxes were examined using the standard artificial digestion method. A simultaneous serological study, extended to include 25239 sera from sows provided by a Swiss pig serum bank, was also undertaken. The results of both studies support the conclusion that Trichinella spp. do not occur within the domestic pig population in Switzerland. Among the fox population, Trichinella was detected in four (0.9%) of the animals tested using the digestion method, and Trichinella britovi was identified as the infecting species by RAPD fingerprint analyses. PMID- 9404847 TI - A survey on resistance to anthelmintics in sheep stud farms of southern Brazil. AB - A survey to investigate the status of anthelmintic resistance in 29 sheep studs in southern Brazil was conducted from March 1992 to December 1993. Compounds from three drug families (macrocyclic lactone, levamisole and benzimidazole) were evaluated concurrently on 22 of the 29 studs. On seven of these properties, resistance to all three families was declared or suspected; at 15 of the 22 studs, ivermectin was the only compound found to be effective in reducing faecal egg counts. Resistance to levamisole was detected on 22 of the 23 studs where it was evaluated and was suspected in the remaining one. The position of benzimidazoles was similar, resistance being declared or suspected on all 28 studs where they were tested. Results of larval cultures indicated that Trichostrongylus, Ostertagia and Haemonchus were the most prevalent nematode genera in the survey, with Trichostrongylus and Haemonchus being the genera associated with anthelmintic resistance. PMID- 9404848 TI - Prophylactic use of ivermectin against cattle myiasis caused by Cochliomyia hominivorax (Coquerel, 1858). AB - The prophylactic efficacy of ivermectin against navel or scrotal myiasis in calves was evaluated in eight trials in Argentina and Brazil. In two trials, calves were injected subcutaneously with ivermectin at a dosage of at least 200 microg kg(-1) within 24 h of birth. In the other six trials, two with two-month old calves and four with four-month-old or older calves, all calves were treated with ivermectin at a dosage of at least 200 microg kg(-1) immediately after castration. In all trials, calves were maintained together on pasture and naturally exposed to Cochliomyia hominivorax. Navel and scrotal wounds were examined for myiasis daily for at least 14 days. Incidence of navel and scrotal myiasis was significantly lower (P < 0.01) in treated calves than in control calves. PMID- 9404849 TI - Altered distribution and expression of protein tyrosine phosphatases in normal human skin as compared to squamous cell carcinomas. AB - Amounts and subcellular localizations of 4 protein tyrosine phosphatases (PTPs) were compared in cultured normal human keratinocytes, an immortalized keratinocyte cell line, and 2 squamous cell carcinoma (SCC) lines. Cellular localizations for PTPs were determined in biopsies of normal human skin and SCCs. Compared to normal keratinocytes, SCC cell lines had higher levels of PTP-1B and T-cell PTP and comparable levels of PTP-1C or PTP-1D. The subcellular localization of each PTP was similar in the 3 types of keratinocytes with PTP-1B localizing to the endoplasmic reticulum, T-cell PTP exclusively found in the nucleus, PTP-1C localized to the plasma membrane, cytosol and nucleus, and PTP-1D present in both cytosol and nucleus. Compared to normal skin, immunoreactive PTP 1B was markedly increased in the invasive margins of SCCs while T-cell PTP was generally increased in tumors. PTP-1C immunostaining varied between cells with no obvious difference between normal and neoplastic tissues. The intensity and distribution of immunoreactive PTP-1D varied greatly between cells within tumors. These differences in amounts and in cellular and subcellular localization of these PTPs, especially those differences in invasive margins of SCCs, may reflect the diverse roles these PTPs play in the proliferation and invasive potential of neoplastic keratinocytes. PMID- 9404850 TI - Differential expression of metallopanstimulin/S27 ribosomal protein in melanocytic lesions of the skin. AB - We have previously shown that human metallopanstimulin (MPS-1) is a ubiquitous 9.4-kDa multifunctional ribosomal S27/nuclear "zinc finger" protein which is expressed at high levels in a wide variety of cultured proliferating cells and tumor tissues, including melanoma. In the present study, we have examined the expression of the MPS-1 protein in various types of human benign and malignant melanocytic lesions of the skin. The expression of the MPS-1 protein was studied by immunohistochemistry using specific anti-MPS-1 antibodies. We found that in benign nevi, the staining is weak and in a gradient; most often, only type A melanocytes stain positive. The B and particularly the C types are negative. Remarkably, congenital nevi show a similar gradient staining of regular benign nevi, but in addition one example showed intensely positive dermal nodules adjacent to areas of negative melanocytes. In melanomas, the staining patterns for MPS-1 are more complex. While some melanomas stain evenly and intensely positive, others have remarkably variable expression of MPS-1. The scattered melanocytes migrating to the upper layers of the epidermis are usually intensely positive. In summary, benign lesions stain in an orderly pattern with staining gradients that correlate with the cellular differentiation of the nevi. Malignant melanomas have an erratic, often intense staining that also correlates with the disorderly growth of these neoplasms. These differential results indicate that the MPS-1 antigen is a useful marker for melanocytic lesions at the immunohistochemical level. PMID- 9404851 TI - The immunofluorescent profile of dermatomyositis: a comparative study with lupus erythematosus. AB - We have demonstrated a role for microvascular injury mediated by the membrane attack complex of complement (C5b-9) in the genesis of cutaneous lesions of dermatomyositis (DM) (1). The purpose of this study is to revisit the immunofluorescent (IF) profile of DM, to further investigate the role of C5b-9 in the pathogenesis of cutaneous lesions, and to see if any features of the IF profile reliably distinguish DM from LE. Lesional skin biopsies from 24 patients with clinical findings characteristic of DM were received in formalin and in Michel's transport medium. Conventional light microscopy, and IF studies with antibodies monospecific for IgG, IgA, IgM, C3, fibrin and C5b-9 were performed. The control group comprised biopsies from 31 patients with well-documented LE. A positive lupus band test (LBT) correlated highly with a diagnosis of LE, with a sensitivity of 64.5% and a specificity of 95.6% (p=0.001). The LBT was most sensitive in the setting of DLE and SLE and was least sensitive in the setting of SCLE. The finding of vascular C5b-9 deposition correlated with a diagnosis of DM versus LE (p=0.001) although the false positive rate was 21.4%. The false negative rate was reduced when vascular C5b-9 was seen in the absence of antibodies to Ro, La, or RNP. While a negative LBT correlated with a diagnosis of DM (p=0.001), the specificity was only 64.5%. However, when it was seen in concert with C5b-9 along the DEJ, specificity was increased to 80.6% (p=0.001). The presence of C5b-9 in vessels and along the DEJ in concert with a negative LBT was predictive of DM (p=0.001) with a specificity of 93.5%, sensitivity of 78.3%, a false positive rate of 10% and a false negative rate of 14.7%. The combination of a negative LBT, vascular C5b-9 deposition and negative serology for Ro, La, and RNP was a predictor of DM versus LE with a sensitivity of 90.5%, a specificity of 96.8%, a false positive rate of 5% and a false negative rate of 6.2% (p=0.001). The IF profile of DM in lesional skin comprises a negative LBT, deposition of C5b-9 within vessels and along the DEJ, and variable keratinocyte decoration for IgG and C5b-9. The most statistically powerful predictor of DM is the combination of a negative LBT with vascular C5b-9 deposition and negative serology for antibodies to Ro, La, Sm, and RNP. Demonstration of a negative LBT in all but 1 case of DM suggests that the DEJ is not a primary site for antigen antibody interaction. We postulate that the aforementioned IF findings reflect humorally mediated injury of endothelium and keratinocytes, effected by C5b-9. PMID- 9404852 TI - Cutaneous lupus mucinosis: a review of our cases and the possible pathogenesis. AB - Cutaneous lupus mucinosis (CLM) is a rare variant of lupus erythematosus eruptions. Our 5 cases with CLM were reviewed. All were men with systemic lupus erythematosus (SLE). CLM occurred as asymptomatic cutaneous papules, nodules, or plaques on the trunk, upper and lower extremities, and face. Histopathology of CLM mainly revealed abundant mucin deposits among splayed collagen bundles throughout the dermis. However, some CLM lesions showed discoid lupus erythematosus-like epidermal and dermal changes and/or lupus profundus. Vasculitis was also revealed in the CLM lesions of 2 cases. The pathogenesis of CLM may be closely related to its two important features, the male preponderance and the association with SLE. Vasculopathy may also be involved in the development of CLM. PMID- 9404853 TI - Absence of human herpesvirus 8 and Epstein-Barr virus genome sequences in cutaneous epithelial neoplasms arising in immunosuppressed organ-transplant patients. AB - Recent studies have implicated herpesvirus 8 and Epstein-Barr virus in the development of cutaneous malignancies in immunosuppressed patients. In order to examine the strength of this association, we examined 37 malignant, pre-malignant and benign cutaneous epithelial neoplasms removed from immunosuppressed organ recipients for the presence of human herpesvirus 8 and Epstein-Barr viral genome sequences using polymerase chain reaction (PCR) and in situ hybridization. We examined 2 actinic keratoses, 1 benign keratosis, 11 invasive squamous cell carcinomas, 17 squamous cell carcinomas in situ and 6 basal cell carcinomas. We also examined 4 basal cell carcinomas, 1 invasive squamous cell carcinoma and 3 squamous cell carcinomas in immunocompetent hosts. In contrast to findings reported by other investigators, we were unable to detect viral genome sequences in any of the biopsies examined. Our findings suggest that human herpesvirus 8 and Epstein-Barr virus likely do not play an etiologic role in cutaneous epithelial oncogenesis in immunocompromised patients. PMID- 9404854 TI - Papillary eccrine adenoma: immunohistochemical studies of keratin expression. AB - Despite various studies, there are serious disagreements about the cellular differentiation of papillary eccrine adenoma. In the present study, 2 specimens of papillary eccrine adenoma were analyzed by immunohistochemical techniques, using a panel of monoclonal antibodies against keratins, to elucidate its differentiation. Histopathologically, the tumor was composed of multiple tubular structures lined by two or more layers of epithelial cells. The luminal cells of the tubules were flattened or cuboidal. The former were noted in large dilated tubules. The latter were usually observed in small-to-moderate-sized tubules, and formed intraluminal papillary projections in some tubules. Immunohistochemically, there were two kinds of cuboidal cells in the luminal layers of the tubules. Most of the large dilated tubules and some of the small-to-moderate-sized tubules expressed immunophenotypes similar to those of the eccrine dermal duct. The other tubular structures, including the small tubules resembling those of syringoma, expressed immunophenotypes similar to those of the transitional portions between the dermal ducts and the secretory segments of eccrine glands. From the above comparative studies, papillary eccrine adenoma is considered to differentiate towards the dermal duct and the transitional portions between the dermal ducts and the secretory segments of eccrine glands. PMID- 9404855 TI - Glomangiosarcoma of the lower limb: a case report with a literature review. AB - Glomangiosarcoma (GS) is a very unusual but morphologically distinctive tumor of soft tissue. We report a case of GS which occurred in the subcutaneous tissue of a 56-year-old man. Microscopically, a typical glomus tumor surrounded a central area of sarcoma. Immunohistochemical stains were performed, and there was a strong positive reaction for vimentin, muscle-specific actin and smooth muscle actin. We also review the clinicopathologic findings of the 9 GS that have previously been reported. PMID- 9404856 TI - Perineurial cell tumor (perineurioma) with granular cells. AB - A form of benign cutaneous tumor with perineurioma findings and with the presence of associated granular cells is described. The two cases studied consisted of whorls made up of a high number of circumferentially arranged flattened cells, with perineurial characteristics, including bipolar cell processes, pinocytotic vesicles, a basal lamina, a positive immunoreactivity for EMA, and absence of immunostaining for S-100 protein. The granular cells, enclosed within the whorls, contained densely packed vesicles, particles with an apparently solid core, as well as membrane-limited vacuoles with disintegrating cellular organelles and electron-dense amorphous material. While failing to demonstrate any immunoreactivity for EMA, the granular cells showed positivity for S-100 protein, which supports their Schwann-cell origin. Due to its morphological and immunohistochemical characteristics, this peculiar form of tumor can be considered as a perineurioma with perineurial cell whorls and granular cell changes occurring in associated Schwann cells at the center of the whorls. PMID- 9404857 TI - Expression of HMB45 antigen in spindle cell melanoma. PMID- 9404858 TI - Inter-relationship between muscle morphology, mechanical output and electromyographic activity during fatiguing dynamic knee-extensions in untrained females. AB - Changes in mechanical performance and electromyographic activity during fatiguing dynamic knee-extensions were evaluated with respect to muscle fibre type composition of the vastus lateralis muscle in nine sedentary female [23 (3) years] volunteers. The subjects performed 150 repetitive maximum knee-extensions using a Cybex dynamometer at 1.57 rad x s(-1). EMG activity was recorded from the vastus lateralis, the vastus medialis and the rectus femoris muscles. For each contraction, mean power frequency (MPF) and the root mean square (RMS) of the EMG were calculated, simultaneously with the peak torque (PT), contractional work (CW) and the mean power (MP). The MPF showed an initial decrease followed by a stable phase. The RMS increased during the initial seven contractions, after which a period of variability was displayed until about the 60th contraction. At the plateau level (last 50 contractions) the relative RMS values were not significantly different from the initial values. The PT, CW and MP increased during the initial five to ten contractions, after which a two-phase pattern was displayed, with a gradual decline followed by a stable phase. The absolute plateau level of MPF for the vastus lateralis muscle showed a significant negative correlation with the area percentage of type-1 fibres (r = -0.71). Significant correlations were also demonstrated to occur between the absolute plateau levels of PT, CW and MP and the relative proportion of type-1 fibres (r = 0.80, r = 0.82 and r = 0.82 respectively). Thus, in female subjects the mechanical performance and the MPF during fatigue are at least partly determined by muscle morphology. PMID- 9404859 TI - Effect of endurance training on pancreatic enzyme activity in rats. AB - The effect of chronic exercise on pancreatic enzyme activity and basal pancreatic secretion was investigated in rats. Male Wistar rats were divided into three groups of ten rats each. In a trained (T) group, the animals were exercised on a treadmill at 35 m x min(-1) for 60 min, 5 days x week(-1). A free-fed control (C) group and a pair-fed control (PFC) group were kept sedentary. Food intake in the PFC group was restricted to the T group levels. After 6 weeks, pancreas wet mass per unit of body mass was significantly larger in the T group in comparison to the C and PFC groups. Protein content, and amylase and lipase activities of the pancreas were significantly higher in the T group in comparison to the C and PFC groups. Basal amylase but not bile-pancreatic juice volume was higher in the T group than in the other two groups. There were no significant differences between groups C and PFC in any of the above parameters. These results would suggest that pancreatic enzyme synthesis and basal secretion are accelerated with physical endurance training. This adjustment would be a beneficial adaptation to chronic endurance exercise, which requires a large energy supply from food. PMID- 9404860 TI - Effects of oxygen fraction in inspired air on force production and electromyogram activity during ergometer rowing. AB - Six male rowers rowed maximally for 2500 m in ergometer tests during normoxia (fractional concentration of oxygen in inspired air, F(I)O2 0.209), in hyperoxia (F(I)O2 0.622) and in hypoxia (F(I)O2 0.158) in a randomized single-blind fashion. Oxygen consumption (VO2), force production of strokes as well as integrated electromyographs (iEMG) and mean power frequency (MPF) from seven muscles were measured in 500-m intervals. The iEMG signals from individual muscles were summed to represent overall electrical activity of these muscles (sum-iEMG). Maximal force of a stroke (Fmax) decreased from the 100% pre-exercise maximal value to 67 (SD 12)%, 63 (SD 15)% and 76 (SD 13)% (P < 0.05 to normoxia, ANOVA) and impulse to 78 (SD 4)%, 75 (SD 14)% and 84 (SD 7)% (P < 0.05) in normoxia, hypoxia and hyperoxia, respectively. A strong correlation between Fmax and VO2 was found in normoxia but not in hypoxia and hyperoxia. The mean sum-iEMG tended to be lower (P < 0.05) in hypoxia than in normoxia but hyperoxia had no significant effect on it. In general, F(I)O2 did not affect MPF of individual muscles. In conclusion, it was found that force output during ergometer rowing was impaired during hypoxia and improved during hyperoxia when compared with normoxia. Moreover, the changes in force output were only partly accompanied by changes in muscle electrical activity as sum-iEMG was affected by hypoxic but not by hyperoxic gas. The lack of a significant correlation between Fmax and VO2 during hypoxia and hyperoxia may suggest a partial uncoupling of these processes and the existence of other limiting factors in addition to VO2. PMID- 9404861 TI - Effects of various beverages on the hormones involved in energy metabolism during exercise in the heat in previously dehydrated subjects. AB - The objective of our study was to examine the effects of beverage content on hormone responses involved in fuel substrate metabolism (catecholamines, insulin and glucagon) in previously dehydrated subjects exercising at a moderate intensity in the heat. Six healthy men walked for 60-min on five occasions at 50% maximal oxygen uptake in a warm environment (dry bulb temperature 35 +/- 0.2 degrees C, relative humidity 20%). On each occasion, the subjects were dehydrated before exercise (loss of 2% body mass) by passive controlled hyperthermia, which led to a reduction in plasma volume (PV) of about -5% to -9%. In one session, the subjects exercised without rehydration (Dh). In the other sessions, four beverages (650 ml) were given just before the exercise: mineral water (W), a 60 g x l(-1) glucose and 1.2 g x l(-1) NaCl solution (GS), a 60 g x l(-1) maltodextrin solution, and a 60 g x l(-1) maltodextrin and 1.2 g x l(-1) NaCl solution. Compared to Dh and W, carbohydrate supply with or without NaCl induced a higher glycaemia (P < 0.05), a reduced increase in plasma adrenaline concentration (P < 0.05) and a higher plasma insulin concentration (P < 0.05), which lowered plasma free fatty acids and glycerol concentrations (P < 0.05). The lesser increase in plasma noradrenaline concentrations observed during GS compared to Dh and W sessions can be explained by a larger correction in PV which might have induced better haemodynamic conditions. However, the increase in plasma glucagon with carbohydrate supply--compared to Dh and W (P < 0.05)--remains unexplained. PMID- 9404862 TI - Relative exercise intensity of long-distance marching (120 km in 4 days) in 153 subjects aged 69-87 years. AB - The aim of this study was to determine the relative exercise intensity (oxygen uptake during the march/maximal oxygen uptake, VO2march/VO2max) during a long distance march in subjects or over 70 years of age. Secondly, the effect of hypertension, cardiovascular and pulmonary diseases on the relative exercise intensity was evaluated. One hundred and fifty-three subjects, 97 men aged 76.7 (4.6) years and 56 women aged 72.8 (3.6) years who completed the 1993 Nijmegen day long-distance march (30 km x day(-1) on 4 consecutive days) participated in the study. Oxygen uptake (VO2) during walking at different velocities (v) was measured in a subgroup of nine men and nine women, selected randomly from the population under study. With these data, regression equations describing the relationship between VO2 and v were made. VO2march was estimated with the obtained regression equations from an average of the v(march) measured in all participants. VO2max was determined using incremental cycle ergometry in all subjects. VO2march was 13.7 (1.8) ml x kg(-1) x min(-1) in men and 15.2 (1.3) ml x kg x min(-1) in women at a mean v of 5 km x h(-1) in both sexes. This corresponded to 52% of VO2max in men and 63% in women. In both sexes subjects with cardiovascular and/or pulmonary diseases walked at a slower v and thus lower VO2march compared to subjects without these diseases. Due to the lower VO2max in subjects with these diseases there was no difference in the relative exercise intensity between the groups. A multiple linear regression analysis showed that and not age on the prevalence of hypertension, cardiovascular and/or pulmonary that VO2max was the most important predictor of the variance in self-selected v(march). This study demonstrates that these active people aged over 70 years could maintain a high relative exercise intensity during endurance walking on 4 subsequent days. Furthermore, it shows that the relative exercise intensity of marching is within the range recommended for improving fitness and reducing the risk of cardiovascular diseases. Finally, these results demonstrate that VO2max has a more important influence on performance than does age or chronic diseases in active elderly people. PMID- 9404863 TI - Dehydration in soldiers during walking/running exercise in the heat and the effects of fluid ingestion during and after exercise. AB - The aim of this study was to examine whether ingesting water alone, or dextrose (7.5 g x 100 ml(-1)) with electrolytes, or fructose/corn solids (7.5 g x 100 ml( 1)) (400 ml every 20 min) would reduce the perceived exertion associated with 16 km (3 h) walking/running in the heat compared with that perceived during exercise with no fluid intake. Perceived exertion was assessed at 1-h intervals during exercise. Blood samples, required for analysis of blood glucose, plasma sodium, plasma osmolality and plasma volume, were obtained prior to exercise and at 1-h intervals during the exercise; further samples were obtained 1-h intervals for 3 h following the exercise. Drinking fluids at regular intervals reduced the level of perceived exertion. In the test during which no fluid was ingested, body mass decreased by 4.9 (0.4) kg [mean (SEM)], but decreased less with ingestion of either the dextrose/electrolytes or fructose/corn solids solutions, or water alone [1.3 (0.2) kg, 1.6 (0.3) kg and 2.0 (0.1) kg, respectively]. Plasma volume fell by 17% when taking no fluid, but fell less when ingesting fluids. Blood glucose fell significantly (P < 0.01) when taking no fluid and rose to 8.4 (1.3) mmol x l(-1) (P < 0.001) and 6.8 (1.1) mmol x l(-1) (P < 0.01) with ingestion of the dextrose/electrolytes or fructose/corn solids solutions, respectively. Urine output was greater with ingestion of water than with any of the other drinks. Six subjects experienced fatigue during exercise with no fluid and failed to complete the exercise. These results suggest that fatigue was caused by several interacting factors: a fall in blood glucose and plasma volume, dehydration, and neuroglycopenia. Taking fluids during exercise reduced the strain and the rating of perceived exertion; this was better achieved by ingesting a dextrose/electrolytes solution. PMID- 9404864 TI - Accumulated oxygen deficit measurements during and after high-intensity exercise in trained male and female adolescents. AB - The purpose of this study was to compare accumulated oxygen deficits and markers of anaerobic metabolism [plasma ammonia (NH3) and lactate (La-) concentrations] in anaerobically trained male [n = 8, age 14.8 (0.5) years; maximal oxygen consumption VO2max 61.74 (2.23) ml x kg(-1) x min(-1)] and female [n = 8, age 14.5 (0.2) years; VO2max 49.62 (3.52) ml x kg(-1) x min(-1)] adolescents. The exercise protocol consisted of runs to exhaustion at speeds predicted to represent 120% and 130% of VO2max. Arterialised blood samples were obtained from a pre-warmed hand via a catheter inserted into a forearm vein. Samples were taken at rest and after 1, 3, 5, 7, 10, 15 and 20 min of recovery. The high-intensity exercise resulted in mean accumulated oxygen deficits that were less (P < 0.05) in females (52.3 ml x kg(-1)) than in males (68.6 ml x kg(-1)). Lower (P < 0.05) plasma concentrations of NH3 and La(-1), and a higher pH were evident in females compared with males during various stages of the 20-min recovery period. The increase in anaerobic performance in the male adolescent athletes when compared with their female counterparts was associated with an increased plasma concentration of selected plasma and blood metabolites. The observed results may reflect well-established differences between the sexes in the morphology and metabolic power of muscle. PMID- 9404865 TI - A new method for data presentation in incremental cardiorespiratory exercise testing. AB - In incremental cardiopulmonary exercise testing, the averaging of data is usually performed to provide group mean data for statistical purposes. They are usually presented as averaged maximum values, or as averaged data at different exercise levels. However, during incremental exercise testing the change in metabolic status may vary between subjects, thus averaging data may not classify the metabolic status accurately. We present an averaging method using a segmented ordinal scale based on individual maximal work performance and the anaerobic threshold (AT). Individual exercise data are grouped into ten classes ranging from unloaded exercise to maximal exercise. The classes are defined in relation to the AT, resulting in an ordinal scale of four classes for exercise data below the AT, one class at the AT and five classes beyond the AT. Resting and unloaded pedalling are treated as separate classes. For evaluation, this method of classification is compared to one based on an absolute scale of oxygen uptake (Cabs) and to another based on a relative scale in 10% steps of maximal oxygen uptake (Crel). Ten healthy male subjects (mean age 23.3 years) performed a ramp cycle ergometer test. When using the Cabs classification method for mean data averaging, mean values for performance at high-intensity exercise were calculated using data from only two of the ten subjects because of variations in individual work capacity. In addition, the AT data were distributed across four classes, thus anaerobic and aerobic exercise data were mixed. Using the Crel classification method enabled data for all ten subjects to be included in the calculation of every data point, but the AT values were still distributed across three classes, resulting in the mixing of anaerobic and aerobic exercise data. However, using the segmented ordinal scale method of classification enabled data from all ten subjects to be included in the calculation of all data points, and it permitted the grouping of the AT values into one class. Thus, this latter method more accurately represents the data of the whole group under study and it allows the metabolic status of the subjects to be taken into consideration. PMID- 9404866 TI - The effect of body temperature on the hunting response of the middle finger skin temperature. AB - The relationship between body temperature and the hunting response (intermittent supply of warm blood to cold exposed extremities) was quantified for nine subjects by immersing one hand in 8 degree C water while their body was either warm, cool or comfortable. Core and skin temperatures were manipulated by exposing the subjects to different ambient temperatures (30, 22, or 15 degrees C), by adjusting their clothing insulation (moderate, light, or none), and by drinking beverages at different temperatures (43, 37 and 0 degrees C). The middle finger temperature (Tfi) response was recorded, together with ear canal (Tear), rectal (Tre), and mean skin temperature (Tsk). The induced mean Tear changes were -0.34 (0.08) and +0.29 (0.03) degrees C following consumption of the cold and hot beverage, respectively. Tsk ranged from 26.7 to 34.5 degrees C during the tests. In the warm environment after a hot drink, the initial finger temperature (T(fi,base)) was 35.3 (0.4) degrees C, the minimum finger temperature during immersion (T(fi,min)) was 11.3 (0.5) degrees C, and 2.6 (0.4) hunting waves occurred in the 30-min immersion period. In the neutral condition (thermoneutral room and beverage) T(fi,base) was 32.1 (1.0) degrees C, T(fi,min) was 9.6 (0.3) degrees C, and 1.6 (0.2) waves occurred. In the cold environment after a cold drink, these values were 19.3 (0.9) degrees C, 8.7 (0.2) degrees C, and 0.8 (0.2) waves, respectively. A colder body induced a decrease in the magnitude and frequency of the hunting response. The total heat transferred from the hand to the water, as estimated by the area under the middle finger temperature curve, was also dependent upon the induced increase or decrease in Tear and Tsk. We conclude that the characteristics of the hunting temperature response curve of the finger are in part determined by core temperature and Tsk. Both T(fi,min) and the maximal finger temperature during immersion were higher when the core temperature was elevated; Tsk seemed to be an important determinant of the onset time of the cold-induced vasodilation response. PMID- 9404867 TI - Physiological and biomechanical responses during treadmill walking with graded loads. AB - Eleven healthy men [mean (SD) for age, height, body mass and maximum oxygen consumption: 25.1 (3.0) years, 1.79 (0.06) m, 78.2 (10.5) kg and 56.9 (7.1) ml x kg(-1) x min(-1), respectively) completed two treadmill walking tests at their self-selected velocity while bilaterally carrying 15-kg and 20-kg loads (in a boxed container) for 4 min in front of the body. Each handle of the boxed container was fitted with a load cell so as to allow quantification of the load supported by each hand during load carriage. During the tests, oxygen uptake (VO2), heart rate (HR), and blood pressure (BP) were monitored using standardized procedures, and cardiac output (Qc) was measured using the carbon dioxide rebreathing method. Stroke volume (SV), arterio-venous oxygen difference (C(a v)O2), rate pressure product (RPP) and total peripheral resistance (TPR) were calculated from the above measurements. The results showed that the two extremities sustained approximately 60% to 70% of the total load, with the balance being supported by the body. Significant increases (P < 0.05) in VO2, HR, Qc, and mean BP were observed during both of the load carriage walks compared to unloaded walking. However, SV, C(a-v)O2, RPP and TPR were unchanged (P > 0.05) during load carriage. Although VO2 was significantly higher during the 20-kg load carriage walk, no significant differences were observed between the two loads for any of the cardiovascular responses monitored. Contrary to our hypothesis, these results suggest that increasing the load from 15 kg to 20 kg during treadmill walking does not significantly increase the cardiovascular stress that occurs in healthy subjects. PMID- 9404868 TI - Thermoregulatory responses of paraplegic and able-bodied athletes at rest and during prolonged upper body exercise and passive recovery. AB - The thermoregulatory responses of ten paraplegic (PA; T3/4-L4) and nine able bodied (AB) upper body trained athletes were examined at rest and during prolonged arm-cranking exercise and passive recovery. Exercise was performed for 90 min at 80% peak heart rate, and at 21.5 (1.7) degrees C and 47.0 (7.8)% relative humidity on a Monark cycle ergometer (Ergomedic 814E) adapted for arm exercise. Mean peak oxygen uptake values for the PA and AB athlete groups were 2.12 (0.41) min(-1) and 3.19 (0.38) l x min(-1), respectively (P<0.05). At rest, there was no difference in aural temperature between groups [36.2 (0.4) degrees C for both groups]. However, upper body skin temperatures for the PA athletes were approximately 1.0 degrees C warmer than for the AB athletes, whereas lower body skin temperatures were cooler than those for the AB athletes (1.3 degrees C and 2.7 degrees C for the thigh and calf, respectively). Upper and lower body skin temperatures for the AB athletes were similar. During exercise, blood lactate peaked after 15 min of exercise for both groups [3.33 (1.26) mmol x l(-1) and 4.30 (1.03) mmol x l(-1) for the PA and AB athletes, respectively, P<0.05] and decreased throughout the remainder of the exercise period. Aural temperature increased by 0.7 (0.5) degrees C and 0.6 (0.4) degrees C for the AB and PA athletes, respectively. Calf skin temperature for the PA athletes increased during exercise by 1.4 (2.8) degrees C (P<0.05), whereas a decrease of 0.8 (2.0) degrees C (P<0.05) was observed for the AB athletes. During the first 20 min of recovery from exercise, the calf skin temperature of the AB athletes decreased further [-2.6 (1.3) degrees C; P<0.05]. Weight losses and changes in plasma volume were similar for both groups [0.7 (0.5) kg and 0.7 (0.4) kg; 5.4 (4.9)% and 9.7 (6.2)% for the PA and AB athletes, respectively]. In conclusion, the results of this study suggest that the PA athletes exhibit different thermoregulatory responses at rest and during exercise and passive recovery to those of upper body trained AB athletes. Despite this, during 90 min of arm-crank exercise in a cool environment, the PA athletes appeared to be at no greater thermal risk than the AB athletes. PMID- 9404869 TI - A comparison between laddermill and treadmill maximal oxygen consumption. AB - Maximal O2 consumption (VO2max) is an index of the capacity for work over an 8 h workshift. Running on a treadmill is the most common method of eliciting it, because it is an easy, natural exercise, and also, by engaging large muscle masses, larger values are obtained than by other exercises. It has been claimed, however, that climbing a laddermill elicits a still higher VO2max, probably because more muscle mass is apparently engaged (legs + arms) than on the treadmill (legs only). However, no data in support of this claim have been presented. To see if differences exist, we conducted progressive tests to exhaustion on 44 active coal miners, on a laddermill (slant angle 75 degrees, vertical separation of rungs 25 cm) and on a treadmill set at a 5% gradient. The subjects' mean (range) age was 37.4 (31-47) years, height 174.3 (164-187) cm, body mass 82.2 (64-103) kg. Mean (range) VO2max on the laddermill was 2.83 (2.31 3.64) l x min(-1) and 2.98 (2.03-4.22) l x min(-1) on the treadmill (P < 0.01, Student's paired t-test). Mean (range) of maximal heart rate f(cmax) (beats x min(-1)) on the laddermill and on the treadmill were 181.0 (161-194) and 181.3 (162-195), respectively (NS). Laddermill:treadmill VO2max was negatively related to both treadmill VO2max x kg body mass(-1) (r = -0.410, P < 0.01) and body mass (r = -0.409, P < 0.01). Laddermill:treadmill f(cmax) was negatively related to treadmill VO2max x kg body mass(-1) (r = -0.367, P < 0.02) but not to body mass (r = -0.166, P = 0.28). Our data would suggest that for fitter subjects (VO2max > 2.6 l x min or VO2max kg body mass(-1) > 30 ml x min(-1) x kg(-1)) and/or higher body masses (> 70 kg), exercise on the laddermill is not dynamic enough to elicit a VO2max as high as on the treadmill. For such subjects, treadmill VO2max would overestimate exercise capacity for jobs requiring a fair amount of climbing ladders or ladder-like structures. PMID- 9404870 TI - Effect of short-term creatine supplementation on renal responses in men. AB - There is an increasing utilisation of oral creatine (Cr) supplementation among athletes who hope to enhance their performance but it is not known if this ingestion has any detrimental effect on the kidney. Five healthy men ingested either a placebo or 20 g of creatine monohydrate per day for 5 consecutive days. Blood samples and urine collections were analysed for Cr and creatinine (Crn) determination after each experimental session. Total protein and albumin urine excretion rates were also determined. Oral Cr supplementation had a significant incremental impact on arterial content (3.7 fold) and urine excretion rate (90 fold) of this compound. In contrast, arterial and urine Crn values were not affected by the Cr ingestion. The glomerular filtration rate (Crn clearance) and the total protein and albumin excretion rates remained within the normal range. In conclusion, this investigation showed that short-term oral Cr supplementation does not appear to have any detrimental effect on the renal responses of healthy men. PMID- 9404871 TI - Core outcomes measures for inguinal hernia repair. AB - BACKGROUND: Demands on the medical profession to develop performance measures and demonstrate cost-effectiveness make it imperative that a uniform approach to the measurement of outcomes for common conditions be adopted. We report here on patient acceptance, response rates, and utility of a new set of core outcomes measures for patients with inguinal hernia (IH), which incorporates patient reporting of outcomes. METHODS: Beginning in March 1994, a convenience sample of patients scheduled for IH repair completed a series of questionnaires addressing a range of patient case mix and outcomes dimensions, including demographics, comorbid conditions, SF-36 health status (Medical Outcomes Study 36-item short form health survey), and condition-specific questions, expectations, and responses to the surgical experience before and after operation. Surgical data were abstracted from the medical records. RESULTS: One hundred three patients were entered in the study; 63 completed 2-month reports and 44 completed 6-month reports. Acceptance of the study and response rates were excellent. Differences in health status associated with IH have been identified in two SF-36 domains, and changes in function after repair noted in several others, supporting the applicability of this measure. Outcomes may also differ by type of hernia and type of repair performed. CONCLUSIONS: A core outcomes measurement set for IH that encompasses demographics, comorbidities, health status, expectations, utilization, and condition-specific data provides a portrait of patient outcomes that is useful to providers and patients, and combined with cost and satisfaction data, it can be used for benchmarking and improving surgical care. PMID- 9404872 TI - Communication effectiveness training improves surgical resident teaching ability. AB - BACKGROUND: An important educational objective of academic surgical programs is to train surgical teachers. Whether formal instruction of surgery residents in general principles of teaching has a role in the achievement of this objective is unproven. STUDY DESIGN: We tested whether the teaching ability of surgery residents could be improved by two different interventions: (A) a lecture on communication effectiveness plus home study of their own videotaped lectures and (B) a critical review of their own videotaped lectures with a teaching consultant. Each resident taught four sessions. There was no intervention between sessions 1 and 2; intervention A occurred between sessions 2 and 3; and intervention B, between sessions 3 and 4. Each of the four videotaped sessions was graded for communication effectiveness using a standardized scoring form. RESULTS: There were no significant differences between scores from lectures 1 and 2 (no intervention) or lectures 2 and 3 (intervention A). Intervention B (individualized feedback) resulted in significant improvement in all scores from session 4 compared with sessions 1 and 2: content 3.40 versus 2.98 (p = 0.01), language 3.43 versus 3.22 (p = 0.03), delivery 3.25 versus 2.87 (p = 0.002), and overall 3.43 versus 2.88 (p = 0.002). CONCLUSIONS: Surgical resident teaching ability can be improved by communication effectiveness teaching. Individualized feedback is more effective than a lecture combined with self-study. PMID- 9404873 TI - Surgical treatment of cancer of the thoracic esophagus in association with a major pulmonary operation. AB - BACKGROUND: Pulmonary complications have been a major cause of mortality after operations for cancer of the thoracic esophagus. Although the risk involved in esophagectomy associated with a major pulmonary operation is expected to be high, it has seldom been evaluated on the basis of clinical experience. STUDY DESIGN: Of 408 patients who underwent esophagectomy, 8 had previously undergone major pulmonary operation (7 for tuberculosis and 1 for pulmonary cancer) and 10 underwent concurrent major pulmonary resection (7 for pulmonary invasion of esophageal cancer, 2 for synchronous pulmonary cancer, 1 for extensive bronchiectasia). All patients underwent systematic lymph node dissection for esophageal cancer, except one patient with mucosal cancer. To prevent postoperative complications, the operative approach and dissection procedures for esophageal cancer were modified according to the associated pulmonary operation and the extent of cancer invasion. All thoracotomies for esophagectomy were performed on the same side as the major pulmonary operation. Additional median sternotomy was performed when necessary. In the most recent 8 patients who underwent major pulmonary resection concurrent with esophagectomy, the bronchial stump was covered with a pedicle flap. RESULTS: Of the 18 patients who underwent pulmonary operation, postoperative complications developed in 13 of the 18 object patients, but none was fatal. The 3-year survival rate was 45%. All deaths were caused by esophageal cancer or another cancer. CONCLUSIONS: Aggressive esophagectomy associated with major pulmonary operation is not contraindicated in patients with fair risk conditions. The operative procedures for esophagectomy should be appropriately modified to minimize the effect of the associated pulmonary operation. Special care should be taken with respect to the approach for mediastinal dissection and closure of the bronchial stump. PMID- 9404874 TI - Total thoracic esophagectomy for esophageal cancer. AB - BACKGROUND: Many current methods of esophageal resection have drawbacks that result in inadequate proximal resection, inadequate lymphadenectomy, and difficult gastric and splenic access. We describe a technique that allows reliable and safe access to the chest, abdomen, and neck. STUDY DESIGN: From 1988 to 1995, 113 patients (82 men; mean age 65.3 +/- 4.5 years) with carcinoma of the esophagus or esophagogastric junction (middle third in 34, lower third in 41, and cardia in 38) underwent total thoracic esophagectomy. The histology was adenocarcinoma in 71 (62.8%), squamous cell carcinoma in 32 (28.3%), and undifferentiated carcinoma in 10 (8.9%) of the patients; 57 tumors (50.5%) were stage III. The esophagus and stomach were mobilized through a left thoracoabdominal incision. After completion of the esophageal resection, the fundus of the stomach was sutured to the esophageal stump to allow later delivery of the stomach into the neck. The esophagogastric anastomosis was performed with continuous single-layer absorbable suture through a left oblique cervical incision. RESULTS: The mean duration of the operation was 309.2 +/- 47.9 minutes. Hospital stay ranged from 5 to 49 days (median, 12 days). The perioperative mortality rate was 4.4%. Anastomotic leak occurred in six patients (5.3%), one of whom died. The proximal resection margin was microscopically free of tumor in all cases, and with a minimum followup period of 18 months, there has been no anastomotic recurrence in any patient. Actuarial survival at 1 year was 63.4% +/- 4.9%, at 3 years 41.4% +/- 5.9%, and at 5 years 22.7% +/- 6.3%. CONCLUSIONS: Total thoracic esophagectomy through the left chest with a separate left cervical incision allows clear access to the esophagus and stomach and good tumor clearance. This procedure may be performed with a low rate of anastomotic leakage, a very low mortality rate, and no anastomotic tumor recurrence. PMID- 9404875 TI - Routine preoperative "one-shot" intravenous pyelography is not indicated in all patients with penetrating abdominal trauma. AB - BACKGROUND: To determine which patients need a "one-shot" intravenous pyelogram (IVP) before laparotomy for penetrating abdominal trauma. STUDY DESIGN: Over a 15 month period, 240 laparotomies were performed for penetrating trauma at our urban level I trauma center. Prospectively collected data included clinical suspicion of genitourinary injury, results of preoperative IVP, intraoperative findings, and operative decisions influenced by the IVP. RESULTS: Preoperative IVP was performed in 175 patients (73%). Of these, 71 (41%) had suspicion of a renal injury based on the presence of a flank wound or gross hematuria. The IVP was believed to influence operative decisions in six patients, all in this group. Each of these six patients had either a shattered kidney or a renovascular injury and had a nephrectomy performed with the knowledge that a normal functioning kidney was present on the contralateral side. No patient without a flank wound or gross hematuria had an IVP that was judged to be helpful intraoperatively. Preoperative IVP was helpful only in patients with flank wounds or gross hematuria. Nephrectomy was performed in two additional patients who did not undergo IVP, both of whom presented in shock. CONCLUSIONS: Routine preoperative IVP is not necessary in all patients undergoing laparotomy for penetrating trauma. The number of IVPs can be safely reduced by 60% if the indications are narrowed to include only those stable patients with a flank wound or gross hematuria. PMID- 9404876 TI - Rapid detection of acute appendicitis with Tc-99m-labeled intact polyvalent human immune globulin. AB - BACKGROUND: Acute appendicitis remains problematic for emergency clinicians. A rapid and definitive test is needed for detecting acute appendicitis before surgical intervention. The purpose of this clinical trial was to determine the efficacy of Tc-99m-labeled intact polyvalent human immune globulin (Tc-99m IgG) in the evaluation of acute appendicitis. STUDY DESIGN: Thirty-five patients with clinically suspected acute appendicitis were evaluated with Tc-99m IgG. After the intravenous injection of 25 mCi (92.5 MBq) of Tc-99m IgG, anterior flow, single photon emission computerized tomography (SPECT) and planar delayed images of the abdomen were obtained. Any abnormal focal uptake of Tc-99m IgG in the right lower quadrant was considered to be a positive scan. RESULTS: Twenty-one patients with a positive Tc-99m IgG scan underwent laparotomy and were found to have acute appendicitis. Of the 14 patients who had negative scans, 7 underwent surgery. In this series, Tc-99m IgG study yielded 21 true-positive, 12 true-negative, and 2 false-negative results with a sensitivity, specificity, and accuracy of 91%, 100%, and 94%, respectively. The positive and negative predictive values were 100% and 86%, respectively. There were no false-positive results. CONCLUSIONS: Tc 99m IgG scintigraphy can provide the clinicians a simple, rapid, and definitive test for the diagnosis of acute appendicitis. PMID- 9404877 TI - The value of the MACE (Malone antegrade colonic enema) procedure in adult patients. AB - BACKGROUND: We report our experience with the Malone antegrade colonic enema (MACE) procedure in adult patients suffering from urinary incontinence and intractable constipation with or without fecal soiling. STUDY DESIGN: Since June 1990, the MACE procedure was initiated in 4 female and 12 male patients 14-54 years old (mean age, 29.9 years) with different pathologic conditions (myelodysplasia, n = 7; anorectal anomaly, n = 3; spinal cord lesion, n = 4; neuropathic disease of unclear cause, n = 2). Three surgical techniques were used: reversed and in situ appendix and tapered ileum). Complex simultaneous urologic continence procedures were performed in nine patients. Two patients had undergone previous operations in the lower urinary tract. RESULTS: After 6.6 years of followup (average, 41.7 months), eight patients (50%) were still using the MACE successfully. They were completely clean day and night and were relieved of symptoms of constipation. Eleven complications related to the MACE procedure occurred in seven patients (44%). Eight patients abandoned the procedure for various reasons. The failure rate was higher in chronically constipated patients without fecal soiling. CONCLUSIONS: The MACE procedure is associated with a high failure rate when used in adults, but it may be possible to identify a subgroup of patients in whom the procedure could be beneficial. Success would depend on overcoming technical problems and difficulties with patient compliance. PMID- 9404878 TI - A modification of hepatic portoenterostomy (Kasai operation) for biliary atresia. AB - BACKGROUND: Although the proportion of patients with biliary atresia remaining jaundice-free after hepatic portoenterostomy (i.e., Kasai operation) has recently been increasing, in many cases repeated reoperation is required to achieve this result. Also, with assessment of jaundice using 2.0 mg/dL of serum total bilirubin as the cutoff, progressive liver fibrosis has occurred in longterm survivors, making liver transplantation necessary. Reoperations result in difficulty in removing the liver for this purpose and cause an increase in the probability of sequelae. STUDY DESIGN: We have performed a new modification of the Kasai operation on a series of patients using the Cavitron ultrasonic suction aspirator (CUSA) for obtaining persistent biliary drainage. We assessed the results using <1.5 mg/dL of serum total bilirubin as the criterion for jaundice free patients. RESULTS: Since 1988, 39 patients were available for review of their clinical results to evaluate our new modification. Thirty patients (77%) were completely and continuously free of jaundice, without living-related liver transplantation or reoperation, and the maximum level of total bilirubin was <1.1 mg/dL. CONCLUSIONS: Our new approach to Kasai operations using CUSA as an integral aid to freeing the biliary remnants and facilitating enteric anastomosis is effective for persistent and complete disappearance of jaundice, without complicated reconstruction or reoperation, and decreases the need for liver transplantation. PMID- 9404879 TI - Ninety-six five-year survivors after liver resection for metastatic colorectal cancer. AB - BACKGROUND: Studies have consistently confirmed the benefit of liver resection for metastatic colorectal cancer. Few reports, however, have a long enough followup or sufficient 5-year survivors to study the clinical course of patients beyond 5 years. STUDY DESIGN: From July 1985 through December 1991, 456 patients underwent liver resection for colorectal metastases. Ninety-six actual 5-year survivors (21%) were identified and their clinical course retrospectively reviewed. RESULTS: Five-year survivors (n = 96) were more likely to have a Duke's B primary colorectal carcinoma, fewer than four metastatic lesions, unilobar disease, and a negative histologic margin when compared with patients not surviving 5 years (n = 298). Forty-four (46%) of the 96 five-year survivors had a recurrence after hepatectomy. Of these 44, 19 (43%) were rendered disease free after further treatment. Overall, 71 of the 96 five-year survivors were free of disease at last followup. The actuarial 10-year survival of this group was 78%. CONCLUSIONS: Patients that are disease free 5 years after liver resection are likely to have been cured by liver resection. Patients should be aggressively followed for recurrence because of the potential for further treatment and longterm survival. PMID- 9404881 TI - Core outcomes measures for inguinal hernia repair. PMID- 9404880 TI - Diagnosis, management, and outcome of late duodenal complications in portal enteric pancreas transplantation: case reports. AB - BACKGROUND: Enteric drainage (ED) of pancreas allografts is an alternative to the bladder drainage (BD) technique and eliminates unique metabolic complications seen in the BD pancreas transplant recipients. Little longterm data has been reported in ED pancreas transplants. STUDY DESIGN: Of 53 patients who underwent pancreas transplantations performed with ED drainage of the exocrine secretion to a Roux-en-Y limb, who had more than 6 months graft function, four patients were identified with late duodenal segment complications (more than 6 months after transplantation) and are presented as case reports. RESULTS: The duodenal segment complications occurred between 8 and 48 months after simultaneous pancreas-kidney transplantation. Three patients were diagnosed with leakage from the duodenal segment. All were managed operatively. The fourth patient developed a distal stricture of the transplant duodenum occluding the anastomosis between the duodenum and the Roux-en-Y limb and also had a pancreatic pseudocyst. Drainage via a cyst-jejunostomy resulted in graft salvage. The mean followup after operative management of the duodenal-related complications was 15 months (range, 3-24 months). The patient, pancreas and kidney graft survival are 100%. CONCLUSIONS: Late duodenal complications occurred in 8% of pancreas transplant recipients with ED. Operative intervention in all four patients resulted in excellent graft and patient outcome and is recommended for these complications. PMID- 9404882 TI - Proximal esophagectomy without laryngectomy followed by free jejunal transfer for esophageal cancer at the cervicothoracic junction. PMID- 9404883 TI - Infrahepatic terminolateral cavo-cavostomy as a rescue technique in complicated "modified" piggyback liver transplantation. PMID- 9404884 TI - Elective extraperitonealization for sigmoid volvulus: an effective and safe alternative. PMID- 9404885 TI - Atelectasis after abdominal surgery. PMID- 9404886 TI - The impact of obesity on surgical outcomes: a review. PMID- 9404887 TI - Advanced breast biopsy instrumentation. PMID- 9404888 TI - The presence of an intron within the rat gene for selenium-dependent glutathione peroxidase 1 is not required to protect nuclear RNA from UGA-mediated decay. PMID- 9404889 TI - A new cyclophilin and the human homologues of yeast Prp3 and Prp4 form a complex associated with U4/U6 snRNPs. AB - We have purified three new human U4/U6-snRNP proteins from HeLa cells. The three proteins formed a tightly bound complex and behaved as a single species throughout the purification. All three proteins have been identified by peptide sequencing, and full-length cDNA sequences have been obtained for all of them. Two of the proteins are homologues of the Saccharomyces cerevisiae splicing factors Prp3 and Prp4, and the third protein is a cyclophilin. Both the human and S. cerevisiae Prp4 proteins have seven repeats of the WD motif and likely fold into structures very similar to those of the beta subunits of G proteins. The human Prp3 protein is highly basic and is closely related to S. cerevisiae Prp3 only in its carboxyl-terminal half. The human homologues of Prp3 and Prp4 are part of a stable complex in the absence of RNA. The third protein in the complex is a new cyclophilin. Cyclophilins have been proposed to act as chaperones in a variety of cellular processes, and we discuss some possible roles of this U4/U6 snRNP-associated cyclophilin. PMID- 9404890 TI - A conditional lethal yeast phosphotransferase (tpt1) mutant accumulates tRNAs with a 2'-phosphate and an undermodified base at the splice junction. AB - tRNA splicing is essential in yeast and humans and presumably all eukaryotes. The first two steps of yeast tRNA splicing, excision of the intron by endonuclease and joining of the exons by tRNA ligase, leave a splice junction bearing a 2' phosphate. Biochemical analysis suggests that removal of this phosphate in yeast is catalyzed by a highly specific 2'-phosphotransferase that transfers the phosphate to NAD to form ADP-ribose 1"-2" cyclic phosphate. 2'-Phosphotransferase catalytic activity is encoded by a single essential gene, TPT1, in the yeast Saccharomyces cerevisiae. We show here that Tpt1 protein is responsible for the dephosphorylation step of tRNA splicing in vivo because, during nonpermissive growth, conditional lethal tpt1 mutants accumulate 2'-phosphorylated tRNAs from eight different tRNA species that are known to be spliced. We show also that several of these tRNAs are undermodified at the splice junction residue, which is always located at the hypermodified position one base 3' of the anticodon. This result is consistent with previous results indicating that modification of the hypermodified position occurs after intron excision in the tRNA processing pathway, and implies that modification normally follows the dephosphorylation step of tRNA splicing in vivo. PMID- 9404891 TI - RNA promoters located on (-)-strands of a subviral RNA associated with turnip crinkle virus. AB - Satellite (sat-) RNA C, one of the nonessential subviral RNAs of turnip crinkle virus (TCV), is dependent on the TCV-encoded RNA-dependent RNA polymerase (RdRp) for its replication. Earlier work showed that a stem-loop structure at the 3' end of (+)-strand sat-RNA C is required for synthesis of (-)-strands in vitro using a partially purified, template-specific TCV RdRp (Song C, Simon AE, 1995, J Mol Biol 254:6-14). Cis-sequences on (-)-strands of sat-RNA C that can serve as separate promoters in vitro have now been defined. Two promoter sequences are located on (-)-strand sat-RNA C, one comprising 11 bases located near the 3' end, and the other consisting of 14 bases located 41 bases from the 5' end. Both promoter sequences contain multiple consecutive C residues followed by multiple consecutive purines and have no obvious secondary structure, suggesting that, along with hairpin structures, specific primary sequences can be recognized by the TCV RdRp. The 3'-proximal promoter sequence directed synthesis from the 3' terminus using (-)-strand templates with the natural sat-RNA 3' end (AUCCC-3'). When plasmid-derived bases were present at the 3' ends of the templates, both promoter sequences could direct the RdRp to initiate transcription internally at the multiple consecutive C residues within the promoters. This result suggests that multiple consecutive C residues are important for transcription initiation and that natural 3'-end sequences, when located at 3' termini, help the RdRp to initiate at the 3' end of the molecule. PMID- 9404892 TI - Redundant RNA recognition events in bicoid mRNA localization. AB - A cis-acting signal in the 3' UTR of the Drosophila bicoid mRNA directs both the transport of the mRNA from the nurse cells to the oocyte and its anterior localization within the oocyte. Here we demonstrate that the signal mediates redundant RNA recognition events, A and B, that initiate largely overlapping programs of mRNA localization during oogenesis. Recognition event A requires a region encompassing stem-loops IV/V of the predicted secondary structure, and can be eliminated by a single nucleotide mutation. Localization initiated through event B begins slightly later in oogenesis, and requires sequences that have not been narrowly defined. Using forms of the 3' UTR lacking this RNA recognition redundancy, we reexamine the roles of the swallow, staufen, and exuperantia genes, which are all required for normal bicoid mRNA localization. Our results reveal that exuperantia first becomes essential for localization at a time when well-defined microtubule tracks between the nurse cells and oocyte disappear. Thus, exuperantia may specifically facilitate a form of nurse cell-to-oocyte mRNA transport not dependent on the microtubule tracks. PMID- 9404893 TI - The Pumilio protein binds RNA through a conserved domain that defines a new class of RNA-binding proteins. AB - Translation of hunchback(mat) (hb[mat]) mRNA must be repressed in the posterior of the pre-blastoderm Drosophila embryo to permit formation of abdominal segments. This translational repression requires two copies of the Nanos Response Element (NRE), a 16-nt sequence in the hb[mat] 3' untranslated region. Translational repression also requires the action of two proteins: Pumilio (PUM), a sequence-specific RNA-binding protein; and Nanos, a protein that determines the location of repression. Binding of PUM to the NRE is thought to target hb(mat) mRNA for repression. Here, we show the RNA-binding domain of PUM to be an evolutionarily conserved, 334-amino acid region at the carboxy-terminus of the approximately 158-kDa PUM protein. This contiguous region of PUM retains the RNA binding specificity of full-length PUM protein. Proteins with sequences homologous to the PUM RNA-binding domain are found in animals, plants, and fungi. The high degree of sequence conservation of the PUM RNA-binding domain in other far-flung species suggests that the domain is an ancient protein motif, and we show that conservation of sequence reflects conservation of function: that is, the homologous region from a human protein binds RNA with sequence specificity related to but distinct from Drosophila PUM. PMID- 9404894 TI - The La protein in Schizosaccharomyces pombe: a conserved yet dispensable phosphoprotein that functions in tRNA maturation. AB - Most RNA polymerase III transcripts are bound immediately after synthesis by an abundant nuclear phosphoprotein known as the La autoantigen. Experiments performed in the budding yeast Saccharomyces cerevisiae have revealed that binding of the La protein to tRNA precursors is required for the endonucleolytic maturation of the 3' terminus of many tRNAs. In the absence of this protein, the 3' ends of these tRNAs are trimmed by exonucleases (Yoo CJ, Wolin SL, 1997, Cell 89:393-402). Here we report the characterization of the La protein in the fission yeast Schizosaccharomyces pombe. As was described for budding yeast, S. pombe cells lacking the La protein are viable and exhibit alterations in the pathway of pre-tRNA maturation. Introduction of either the human, S. cerevisiae, or S. pombe La protein into these cells restores the detected pattern of tRNA processing intermediates to that of wild-type cells. By performing immunoprecipitations from cells that were metabolically labeled with 32P-orthophosphate, we demonstrate that the S. pombe and S. cerevisiae La proteins, like the human La protein, are phosphorylated in vivo. Thus, although the La protein is dispensable for growth in these yeasts, both the structure of the protein and its function in pre-tRNA maturation have been highly conserved throughout evolution. PMID- 9404895 TI - Identification of a novel, non-snRNP protein complex containing U1A protein. AB - Mouse monoclonal antibodies (MAbs) were generated against Escherichia coli produced U1snRNP-A (U1A) protein. U1A-specific MAbs as well as MAbs that reacted with both U1A and U2snRNP-B" (U2B") were isolated. MAb 12E12 was unique among the characterized MAbs because it failed to immunoprecipitate U1A protein produced by in vitro transcription and translation using rabbit reticulocyte lysates. However, when U1A protein was made using a wheat germ extract, MAb 12E12 could immunoprecipitate U1A quite readily, as did the other MAbs. These data suggest that the MAb 12E12 epitope is masked when U1A is prepared in reticulocyte lysate. Further studies showed that MAb 12E12 recognizes an epitope that is masked when U1A protein is bound to U1 RNA. The unique nature of MAb 12E12 was used to demonstrate that U1A could be immunoprecipitated from whole-cell extracts in a form that was free of U1 RNA and other snRNP components. However, this snRNP-free U1A (SF-A) was found to co-immunoprecipitate with a unique set of non-snRNP proteins. In order to confirm that U1A exists in at least two distinct complexes (snRNP bound and snRNP free), [35S]-labeled nucleoplasmic extracts were analyzed by sucrose density gradient fractionation and immunoprecipitation. MAb 12E12 specifically immunoprecipitated SF-A, which migrated in a novel non-snRNP complex. Specifically, proteins of approximately 58, 59, 63, 65, and 105 kDa co sedimented and co-immunoprecipitated with SF-A. Our data show that a significant portion of the cellular U1A (at least 3% or approximately 30,000 molecules) exists in the nucleoplasm in one or more novel complexes. Our previous studies have demonstrated an effect of purified U1A on polyadenylation of pre-mRNAs and, consistent with this finding, purified antibodies to SF-A significantly diminish polyadenylation in vitro. PMID- 9404896 TI - Both phosphorylation and dephosphorylation of ASF/SF2 are required for pre-mRNA splicing in vitro. AB - The splicing reaction that removes introns from pre-messenger RNAs requires the assembly of the spliceosome on the nascent transcript, proper folding of the substrate-enzyme complex, and finally, two transesterification reactions. These stages in the splicing reaction must require careful orchestration. Here we show data that suggest that the sequential phosphorylation and dephosphorylation of SR proteins mark the transition between stages in the splicing reaction. Many data had already led to the idea that phosphorylation of SR proteins could modulate their activity, when we showed that dephosphorylation of these proteins abrogates their activity in a reaction measuring conversion of pre-spliceosomes to spliceosomes (Roscigno RF, Garcia-Blanco MA, 1995, RNA 1:692-706). Subsequently, Xiao and Manley (1997, Genes & Dev 11:334-344) showed that phosphorylated ASF/SF2, but not mock-phosphorylated ASF/SF2, activates the splicing of HIV tat pre-mRNA in reactions challenged with excess random RNA. Here we confirm and extend these two findings. Phosphorylated ASF/SF2 efficiently complemented an SR protein-deficient HeLa S100 extract in promoting the splicing of an adenovirus-2 derived pre-messenger RNA, whereas unphosphorylated ASF/ SF2 did not. Moreover, we show that, whereas unphosphorylated ASF/SF2 inhibited splicing in HeLa nuclear extracts, phosphorylation of the ASF/SF2 reversed the inhibition and enhanced splicing. We also present data that shows that dephosphorylation of ASF/SF2 is required for the first transesterification reaction once the spliceosome has assembled. Thiophosphorylated ASF/SF2, which cannot be readily dephosphorylated, can promote spliceosome assembly, but cannot promote the first transesterification reaction. These data, together with other observations, indicate for the first time a requirement for SR protein dephosphorylation in pre messenger RNA splicing in vitro. PMID- 9404897 TI - Complex formation of the spinach chloroplast psbA mRNA 5' untranslated region with proteins is dependent on the RNA structure. AB - RNA-protein interactions are part of many regulatory pathways in gene expression. In chloroplasts of higher plants and of green algae, gene regulation by posttranscriptional processes such as differential regulation of mRNA stability and control of translation plays a major role during chloroplast development and light-dependent protein expression. Regulation here is mediated by interactions of RNA-binding proteins with the respective mRNAs. In this work, structural requirements for protein-RNA complex formation between the 5' untranslated region of the spinach psbA mRNA (encoding the D1 protein of photosystem II) and stromal proteins are analyzed. For this, a combination of temperature gradient gel electrophoresis and gel shift analysis is employed to study several variants of the psbA 5' untranslated region. Supported by theoretical interpretation of the data and analysis of the structures by chemical probing, we show that a certain structure of the RNA is necessary for protein complex formation. Already very subtle structural changes within the RNA interfere with binding and thereby with the biological activity of the mRNA. PMID- 9404898 TI - Nucleotide G-1207 of 18S rRNA is an essential component of the human 80S ribosomal decoding center. AB - mRNA analogues-derivatives of oligoribonucleotides consisting of two different codons and bearing an aryl azide group at the 5'-phosphates-were crosslinked to human 80S ribosomes by UV-irradiation of the various model complexes obtained in the presence of the cognate tRNAs. Three sequences, namely pUUUGUU (coding for Phe and Val), pUUCUAAA (first triplet coding for Phe and second being stop codon), and pGUGUUU (coding for Val and Phe), have been used. Sequences of 18S rRNA containing nucleotides crosslinked to the mRNA analogues were examined by hydrolysis with RNase H in the presence of various cDNA probes. Crosslinked nucleotides were identified by primer extension. In all cases, only nucleotide G 1207 (equivalent to G-926 in Escherichia coli 16S rRNA) has been detected as crosslinked. Crosslinking of the mRNA analogues to the large ribosomal subunit was negligible. PMID- 9404899 TI - The deadly quartet--the insulin resistance syndrome. AB - BACKGROUND: Obesity, non-insulin-dependent diabetes mellitus, hypertension, and dyslipidemia (syndrome X, "the deadly quartet") are common metabolic disorders that predispose to early cardiovascular disease. We examine the relationship between insulin resistance and the deadly quartet and address therapeutic implications. METHODS: We review the literature on insulin resistance, using MEDLINE files from 1975 to the present. Fifty references were reviewed. RESULTS: Insulin resistance consists of a cluster of disorders and biochemical abnormalities. We discuss the mechanisms responsible for the defects in insulin mediated glucose utilization, as well as the relation of insulin resistance to obesity, hypertension, and dyslipidemia. We review the current strategies used in light of this pathophysiologic approach. CONCLUSIONS: This extremely common syndrome contributes excessively to mortality and morbidity of millions of Americans and generates enormous costs to the health care system. Better molecular understanding of insulin resistance is leading to improved treatment of all components of the syndrome. PMID- 9404901 TI - Focal reexpansion pulmonary edema after drainage of large pleural effusions: clinical evidence suggesting hypoxic injury to the lung as the cause of edema. AB - BACKGROUND: The purposes of this study were to review possible causes of reexpansion pulmonary edema (RPE) and to attempt to explain atypical distributions of RPE after drainage of large pleural effusions. METHODS: Five patients had focal RPE after routine drainage of large pleural effusions. In these cases, pleural effusion did not completely fill the hemithorax, and part or all of the ipsilateral upper lobe remained aerated. Reexpansion was accomplished by chest tube drainage with -20 cm H2O suction in four cases and by percutaneous needle aspiration without application of negative intrapleural suction in one. RESULTS: In all five cases, RPE developed in the portion of the lung that had been collapsed but did not develop in the portion of the lung that remained aerated. CONCLUSIONS: This suggests that hypoxic injury to the atelectatic lung, rather than mechanical stress, is the most plausible explanation for RPE. PMID- 9404900 TI - Role of hydrolyzed formulas in nutritional allergy prevention in infants. AB - BACKGROUND: In recent years, more and more discussions have arisen with regard to the role of (partially) hydrolyzed formulas as standard feedings for infants with a high risk to have allergy. METHODS: This review is based on an extensive overview of the literature dealing with the subjects of allergy prevention and hydrolyzed formulas. RESULTS: Although breast-feeding should receive absolute priority in the nutrition of infants, the existence of artificial milk formulas as an addition to or replacement of breast milk is a necessity. In high-risk infants with a family history of allergy, we might consider a hypoallergenic formula instead of the classical start formulas to reduce the risk of allergy. From a nutritional point of view, these formulas should only be hydrolyzed as much as necessary. On the other hand, for the treatment of food allergies, the peptides of the semi-elementary infant formulas should be as short as possible. This can, however, have an impact on the nutritional value of the formula. Therefore, a difference is made between partial and complete hydrolysates. CONCLUSION: While a firm recommendation is not yet possible, physicians might consider partial hydrolysate formulas in high-risk infants if parents can afford the higher-cost option. PMID- 9404902 TI - Significance of nondiagnostic fine-needle aspiration of the thyroid. AB - BACKGROUND: Fine-needle aspiration biopsy (FNAB) has been shown to be rapid and cost effective in the evaluation of thyroid nodules. The significance of nondiagnostic (unsatisfactory) FNAB is uncertain, however. METHODS: We reviewed 345 consecutive thyroid FNABs and identified 59 patients with initially unsatisfactory specimens. These patients had follow-up to determine whether their thyroid nodules proved to be malignant. RESULTS: Three patients (5.1%) were found to have organ-confined papillary carcinoma of the thyroid, the largest tumor mass measuring 1.2 cm. Six patients (10.2%) had benign adenomas. CONCLUSIONS: In most cases of initially nondiagnostic FNAB of a thyroid nodule, neoplasia is not found subsequently. A minority of cases may still harbor malignancy. None of our patients in whom repeated FNA was either nondiagnostic or suggestive of benign disease were ultimately found to have a malignancy. PMID- 9404903 TI - Pulmonary function tests: comparison of 95th percentile-based and conventional criteria of normality. AB - BACKGROUND: Although 95th percentile-based normal limits are recommended instead of conventional criteria of normality to guide pulmonary function test (PFT) readings, we have found no objective assessment of how the choice of normal limits might influence PFT interpretation. METHODS: We did a retrospective comparison of PFT readings referenced to conventional criteria of normality versus independent repeat assessments influenced by 95th percentile-based normal limits in 166 veterans. We also conducted a nationwide telephone survey of VA Hospital PFT laboratories. RESULTS: Discordant readings occurred in only 7.2% of 616 individual PFTs; however, these discrepancies could potentially influence at least one component of the PFT report of 26.5% of our subjects. The 95th percentile-based normal limits were used by only 40% of VA PFT laboratories, without relationship to geography or hospital size. CONCLUSIONS: Discrepancies between 95th percentile-based and conventional normal limits can potentially influence PFT readings, and 95th percentile-based criteria are not used in the majority of VA PFT laboratories. PMID- 9404904 TI - Multicenter, randomized study comparing levofloxacin and ciprofloxacin for uncomplicated skin and skin structure infections. AB - BACKGROUND: The fluoroquinolone, levofloxacin, is active against most common pathogens in skin and skin structure infections. METHODS: The efficacy, tolerability, and safety of levofloxacin and ciprofloxacin were compared in a randomized, open-label, multicenter trial of patients with uncomplicated skin and skin structure infections. Of 469 patients treated, 231 received levofloxacin (500 mg qd) and 238 were given ciprofloxacin (500 mg bid). RESULTS: Overall clinical success rates (cured plus improved) for levofloxacin and ciprofloxacin were 98% and 94%, respectively (95% confidence interval [CI], -7.7, 0.7). Overall microbiologic eradication rates by patient were 98% in the levofloxacin group and 89% in the ciprofloxacin group (95% CI, -14.5, -2.7), whereas eradication rates by pathogen were 98% and 90%, respectively (95% CI, -12.6, -3.7). The eradication rate for Staphylococcus aureus was 100% in the levofloxacin group and 87% in the ciprofloxacin group (95% CI, -20.2, -5.1). Treatment-emergent adverse events were comparable, with drug-related adverse events reported in 6% of levofloxacin patients and 5% of ciprofloxacin patients. CONCLUSIONS: Levofloxacin is as effective and safe as ciprofloxacin in the treatment of uncomplicated skin and skin structure infections. PMID- 9404906 TI - Cefixime: an oral option for the treatment of multidrug-resistant enteric fever in children. AB - BACKGROUND: Enteric fever is a serious public health problem in Pakistan, where multidrug-resistant salmonellosis causes enteric fever with increased morbidity and mortality. Costly parenteral therapy and lack of an established safety profile for the use of quinolones in children necessitate evaluation of an oral treatment option. This study is meant to assess the efficacy, safety, and cost effectiveness of an oral third-generation cephalosporin (cefixime) in the treatment of multidrug-resistant enteric fever. METHODS: Between November 1993 and October 1994, 85 patients, 15 years of age or less, with culture-proven enteric fever were randomly assigned to two groups. Group A (n = 41) received cefixime at a dosage of 10 mg/kg to 12 mg/kg per day in two divided doses. Group B (n = 44) received chloramphenicol at a dosage of 100 mg/kg daily in four divided doses. Both groups were treated for 2 weeks. RESULTS: In group A, 95% (39/41) of the patients receiving cefixime responded favorably, whereas in group B, 30% (14/45) responded to chloramphenicol. The 31 patients not cured in group B were then successfully treated with cefixime. Overall, cefixime was well tolerated. Subsequent antibiogram data showed an overall multidrug-resistance rate of 78% (66/85). CONCLUSIONS: Cefixime is a safe, effective, and cheaper oral option for the treatment of multidrug-resistant enteric fever. Further studies are needed, however, to validate this observation. PMID- 9404905 TI - Hypothyroid Graves' disease. AB - BACKGROUND: Spontaneous conversion from hypothyroidism to hyperthyroidism has generally been considered uncommon. METHODS: Values obtained were serum thyroid stimulating hormone (TSH), thyroxine, free thyroxine index, radioactive iodine uptake (RAIU), thyroid-stimulating immunoglobulins (TSI), and thyrotropin-binding inhibitory immunoglobulin (TBII). RESULTS: Five patients spontaneously had a minimum of two cycles in thyroid function with extremes of hypothyroxinemia to hyperthyroxinemia. One patient had four documented cyclic shifts in thyroid status. When measurements were obtained in the hyperthyroid phase, all patients had TSI, increased RAIU, and an undetectable TSH. When measurements were done in the hypothyroid phase, all patients had positive TBII but negative TSI. CONCLUSIONS: Spontaneous reversal of thyroid function may be more common than previously thought. Clinical features associated with lability of thyroid function were abrupt change in goiter size, exaggerated response to therapy, or the presence of TSH-receptor antibodies. PMID- 9404907 TI - Prospective assessment of triage in an urban emergency department. AB - BACKGROUND: This study examined the effectiveness of a triage system based on patient complaints, medical history, vital signs, and triage nurse impression. Measurements included recognizing patients needing admission, in correlating with disposition, and its effectiveness in all age groups. METHODS: Data were collected prospectively on all patients coming to a general emergency department (ED) of an urban teaching hospital from October 1, 1992, through November 30, 1992. Data included assigned triage acuity, disposition waiting time to physician examination, and disposition, as well as return to the ED within 2 weeks. The patients were divided into age groups: 0 to 16 years, 17 years to 25 years, 25 years to 50 years, 50 years to 65 years, and >65 years of age. RESULTS: There were five patients (n = 4,993, 0.4%) who were triaged nonemergently and subsequently admitted. The sensitivity and specificity of an assigned triage 3 acuity assignment in correlating with lack of admission were 99% and 56%, respectively. Mean waiting time to physician examination was 61 +/- 14 minutes for triage 1, 129 +/- 19 for triage 2, and 182 +/- 22 for triage 3. Mean time to admission from sign-in was 246 +/- 10 minutes for triage 1 and 372 +/- 16 minutes for triage 2. CONCLUSIONS: This triage system accurately correlated with disposition and determined waiting time to examination. PMID- 9404908 TI - Safety of carotid endarterectomy associated with small intracranial aneurysms. AB - BACKGROUND: The incidence and outcome of combined carotid artery disease and intracranial aneurysm (ICA) are not well reported. METHODS: Ten patients with combined disease, ICA and symptomatic carotid artery disease, were identified in 209 consecutive angiograms. Five men and five women with a mean age of 68 years and the risk factors of diabetes, hypertension, smoking, cardiac disease, peripheral vascular disease, and hypercholesterolemia formed the basis for this study. RESULTS: Five patients with carotid endarterectomy (CEA) and arterial aneurysms less than 5 mm and five with carotid stenosis and ICA less than 6 mm were treated with Coumadin; one with combined disease left the hospital without treatment; and one with combined disease died preoperatively of a myocardial infarction. One patient with a 2 cm x 3 cm ICA and carotid had both operated on successfully. CONCLUSIONS: In this group of patients, CEAs were done safely in patients with ICAs less than 6 mm. PMID- 9404909 TI - Perioperative transfusion, postoperative infection, and recurrence of head and neck cancer. AB - BACKGROUND: Immunologic effects of perioperative transfusion and postoperative infection have been purported to influence cancer recurrence rates. METHODS: Records of all head and neck cancer patients having surgical extirpation of the primary tumor and/or regional nodes at our institution over a 5-year period were reviewed. Time to recurrence was the outcome measure. All variables were evaluated via univariate analysis using log rank tests, with Cox proportional hazards used for multivariate analyses. RESULTS: Univariate analysis identified the following as potential prognostic factors associated with recurrence: nodal stage, total lymphocyte count, overall stage, amount transfused, occurrence of a transfusion, and the American Society of Anesthesiologists status. Various backward stepwise multivariate regression models showed that neither transfusion nor postoperative infection independently influenced recurrence. However, transfusion of 3 or more units did surface as an independent contributor to recurrence, and in certain subgroups there was a trend toward improved survival for those who had a postoperative infection. CONCLUSIONS: In this series, neither perioperative transfusion nor postoperative infection independently influenced recurrence. PMID- 9404910 TI - Undocumented smokeless tobacco use in a family practice population. AB - BACKGROUND: Smokeless tobacco (ST) use is associated with significant health risks and is increasing in prevalence in the United States. This study examined the prevalence of ST use among patients seen in two family practice centers and the documentation of that ST use in the medical record. METHODS: A survey of 209 patients seen in two family practice centers in northeast Tennessee was done to determine current and former smoking and ST use. Charts of all current ST users were reviewed for additional data. RESULTS: Current ST use was reported by 7.7% of patients. Highest rates were found in middle-aged men (27.8%) and elderly women (14.8%). Physicians rarely recorded ST use as a significant problem in the medical record. In several cases, management of other medical conditions may have been compromised by the physicians' lack of awareness of the ST use. CONCLUSIONS: The use of smokeless tobacco may be an important unrecognized problem in patients seen in family physicians' offices. PMID- 9404912 TI - Recurrence of spondylothoracic dysplasia (Jarcho-Levin syndrome) in a family. AB - We describe a rare occurrence of two consecutive cases of spondylothoracic dysplasia (Jarcho-Levin syndrome) in a nonconsanguineous Hispanic family. Serial measurements of pulmonary function and energy expenditure were useful in one of these infants for assessment of the evolution and severity of restriction of pulmonary function and the determination of timely therapeutic intervention. PMID- 9404911 TI - Premature rupture of membranes at term with an unfavorable cervix: comparison of expectant management, vaginal prostaglandin, and oxytocin induction. AB - BACKGROUND: Our objective was to determine the best treatment for parturients at term with an unfavorable cervix and premature rupture of membranes (PROM). METHODS: In this prospective study, 96 women with PROM and an unfavorable cervix were randomized into one of three treatment groups: oxytocin induction, vaginal prostaglandin E2 gel followed by oxytocin, or expectant management. RESULTS: Length of labor, cesarean section rate, and maternal/neonatal morbidity were not significantly different. In contrast, the interval from PROM until delivery and length of hospital stay were significantly longer in the expectantly managed group than in the other groups. Four of the patients who received expectant management required delivery because of nonreassuring fetal assessments. CONCLUSIONS: Expectant management of PROM at term significantly prolongs hospital stay without decreasing the incidence of abdominal delivery or infectious morbidity. There appears to be potential for cord compression in patients managed expectantly without continuous electronic fetal surveillance. PMID- 9404913 TI - Colonic metastasis from a primary renal leiomyosarcoma: an unusual cause of gastrointestinal hemorrhage. AB - Primary leiomyosarcoma of the kidney is rare. We report the first case of such a tumor that metastasized to the colon and was manifested by gastrointestinal hemorrhage. PMID- 9404914 TI - Airway management in a patient with Hecht's syndrome. AB - Hecht's syndrome, described in 1969, is a rare autosomal dominant phenotype that includes trismus, pseudocamptodactyly, and somewhat short stature. A 5-year-old white boy with Hecht's syndrome and frequent otitis media, nasal obstruction, and sleep apnea is described to illustrate the otolaryngologic manifestations of this syndrome. Airway management in these patients is complicated by trismus, which does not improve after the induction of anesthesia. Mask anesthesia with spontaneous ventilation has been used successfully in these patients but may be difficult in the majority of otolaryngologic procedures. Blind nasotracheal intubation is an alternative, but in this case it was impossible because of adenoid hypertrophy. In this patient, laryngoscopy and orotracheal intubation were done using a Bullard laryngoscope, and transnasal KTP laser adenoidectomy was done to relieve the obstruction and apnea. The details of Hecht's syndrome and its management are presented in this case review. PMID- 9404915 TI - Thrombosis of the deep femoral vein: a potential pitfall of color flow duplex Doppler ultrasonography. AB - This case illustrates a potential pitfall of color flow duplex Doppler ultrasonography with compression in the evaluation of suspected deep venous thrombosis (DVT). Because of its low cost, accuracy, and noninvasiveness, ultrasonography is the appropriate first choice in the evaluation of suspected DVT, but there does exist the possibility of a false-negative examination. Magnetic resonance venography (MRV) should be reserved for cases in which there is a high clinical suspicion for DVT, as well as either morbid obesity that would limit the evaluation of deep pelvic and deep femoral veins or conflicting results of other imaging studies. All cases of suspected thrombosis, including those not adequately evaluated by ultrasonography, can be accurately assessed by MRV, which is not as invasive as standard venography. PMID- 9404916 TI - Acute pansinusitis with bacteremia due to a beta-hemolytic group C streptococcus: Streptococcus milleri. AB - A 41-year-old truck driver had acute onset of weakness, severe headache and pain over the left side of the face, forehead, and orbital area. He was found to have acute pansinusitis. Blood cultures and culture of the sinus drainage yielded beta hemolytic group C streptococcus: Streptococcus milleri. He recovered completely after treatment with cefazolin, surgical drainage and debridement, and outpatient cephalexin therapy. Beta-hemolytic streptococci are uncommon causes, and bacteremia is rare in acute sinusitis. Speciation of the streptococcus is important in determining the epidemiology and clinical spectrum of streptococcal infections. PMID- 9404917 TI - Pneumoappendix after an inversion appendectomy: an unreported radiologic finding. AB - Radiographically detectable gas only occasionally fills the lumen of the vermiform appendix (pneumoappendix), and the diagnostic significance of this gas remains unresolved in the radiologic literature. The following case report is a previously unreported finding of a postoperative plain abdominal radiograph showing a dilated pneumoappendix after an inversion-ligation appendectomy. A brief review of the surgical technique is included to help explain these radiographic findings. PMID- 9404918 TI - Sudden death due to disseminated cryptococcosis in a child with leukemia in remission. AB - Cryptococcus neoformans typically causes an insidious illness with symptoms related to meningitis or to lung involvement. This is the first reported sudden death due to cryptococcosis, which occurred in a child with leukemia that was in remission. The child had suddenly looked seriously ill and cried with abdominal pain and then died within 25 minutes. Disseminated cryptococcal infection of the lungs, heart, and pancreas was an unexpected finding at autopsy. This clinical experience raises the question whether fungal infections should now be considered in immunosuppressed patients who have an apparent septic collapse. PMID- 9404919 TI - Shifting percussion dullness of the chest: a sign of pleural effusion. AB - Early physicians diagnosed pleural effusion by detecting shifting percussion dullness of the chest. However, the lateral decubitus positions have not been routinely used to distinguish this condition from atelectasis or pneumonia. In this report, postural alterations of percussion dullness established the diagnosis of pleural effusion in a 49-year-old patient with a cough. Percussion in the lateral decubitus positions may enhance the value of examinations of the chest. PMID- 9404920 TI - Nephrotoxicity after high-dose carboplatin, etoposide and ifosfamide in germ-cell tumors: incidence and implications for hematologic recovery and clinical outcome. AB - High-dose carboplatin, etoposide and ifosfamide (CEI) is an active chemotherapy regimen (HDCT) in solid tumors and lymphomas. In patients with previous exposure to cisplatin, its nephrotoxicity is dose-limiting. To determine the implications of nephrotoxicity on hematological recovery and clinical outcome, we analyzed 150 consecutive patients with germ cell tumors treated between August 1989 and September 1995 with carboplatin 1500-2000 mg/m2, etoposide 1200-2400 mg/m2, and ifosfamide 0-10 g/m2 followed by either BM or PBPC rescue. Five patients died (3%), three in the context of severe renal toxicity and early multiorgan failure. Overall, acute nephrotoxicity occurred in 43/150 (29%) patients, particularly at doses of carboplatin >1500 mg/m2. Hemodialysis was required in 12/150 (8%) patients, but could be discontinued until discharge in all except two survivors. Nephrotoxicity did not delay hematologic recovery when adjusted for the use of PBPC and hematopoietic growth factors by multivariate analysis, but resulted in higher transfusion requirements, more overall toxicities and a longer hospital stay. There were no differences in the response rates or survival in patients with or without nephrotoxicity. Acute nephrotoxicity is a frequent and clinically relevant complication of CEI in germ cell tumors. The acute side-effects from CEI are reversible in the majority of patients. PMID- 9404921 TI - Autologous peripheral blood stem cell transplantation for acute myelogenous leukemia. AB - The safety and efficacy of myeloablative therapy followed by autologous peripheral blood stem cell transplantation (ABSCT) for acute myelogenous leukemia (AML) were evaluated in 60 patients. Peripheral blood stem cells (PBSC) were collected during recovery after consolidation chemotherapy. High-dose chemotherapy consisting of busulfan (16 mg/kg), etoposide (40 mg/kg), and cytosine arabinoside (3 g/m2 x 4) (BEA regimen) was used for pretransplant conditioning in 13 patients. For the remaining 47 patients, granulocyte colony stimulating factor (G-CSF) was administered concurrently with the BEA regimen during conditioning. Unpurged, cryopreserved PBSC containing a median number of 5.4 x 10(8) MNC/kg or 12 x 10(4) CFU-GM/kg were reinfused at transplantation. The median number of days to granulocytes exceeding 500/microl and last platelet transfusion were 15 (8-44) and 24 (0->180), respectively. The 3-year probabilities of disease-free survival (DFS) and relapse were 78.6 and 21.4% for patients transplanted in first remission, 29.6 and 64.4% for those in second or third remission, and 11.1 and 77.8% for those in relapse, respectively. There were no transplant-related deaths within 100 days of transplantation. Age, disease status at transplantation, and number of induction chemotherapies to first complete remission were risk factors affecting the outcome of ABSCT. These results of ABSCT for AML in first remission warrant a prospective study of ABSCT as post-remission therapy. PMID- 9404923 TI - Mobilization of hematopoietic progenitors in patients with chronic myeloid leukemia. AB - Although the mobilization and harvesting of Philadelphia chromosome-negative progenitors has been proven feasible by a number of groups, it is not clear which techniques should be used with regard to the monitoring of purging and evaluation of stem cell yield. Therefore, we isolated CD34-positive cells from leukapheresis samples of seven CML patients after induction therapy. These cells were analyzed using the colony-forming unit granulocyte-macrophage (CFU-GM) and long-term culture-initiating cell (LTCIC) assays. In addition, we analyzed frequency and clonogenicity of single stem cells using the LTCIC assay at limiting dilution. Individual colonies derived from these assays were subsequently analyzed for the presence of the bcr-abl gene with interphase fluorescence in situ hybridization (FISH) and for the presence of bcr-abl mRNA with RT-PCR. We compared these results with the cytogenetic analysis of the leukapheresis material. Molecular analysis of individual colonies correlated well with the results of cytogenetic analysis. However, in nine out of 18 samples, cytogenetic analysis exclusively demonstrated the presence of either Philadelphia chromosome-positive or -negative cells whereas with the molecular analysis of individual colonies tumor contamination or the presence of residual normal cells could be substantiated. We concluded that molecular analysis of individual colonies should be employed to further optimize induction protocols. With regard to stem cell mobilization we demonstrated that about 67 CFU-GM colonies and clusters were generated by one single LTCIC both for the CML samples and the samples obtained from patients with non-hematologic malignancy. This number is important since until now most centres use a number of four colonies and clusters generated by one LTCIC. Dividing the CFU-GM output in the LTCIC assay by four to determine LTCIC frequency could thus lead to an almost 20-fold overestimation of the LTCIC frequency. It is concluded that stem cell frequency analysis is an important tool with regard to the interpretation of mobilization protocols. PMID- 9404924 TI - High-dose busulphan/melphalan with autologous stem cell rescue in Ewing's sarcoma. AB - Eighteen patients with poor risk Ewing's sarcoma (including 11 patients with metastatic disease at presentation) received consolidation therapy of busulphan and melphalan with autologous stem cell rescue. There were nine females. The median age at diagnosis was 14.2 years (range 2.75-30 years). There was one early death due to cytomegalovirus pneumonitis. One patient developed a single generalised convulsion during busulphan therapy. Severe renal toxicity was not encountered. One patient developed veno-occlusive disease of the liver (VOD) which eventually resolved. With a median follow up of 2 years, 13 patients survive including six with initial metastatic disease. We conclude that high-dose busulphan/melphalan is well-tolerated and should be evaluated for efficacy in a larger series of patients with high risk Ewing's sarcoma. PMID- 9404922 TI - CD34+-selected autologous peripheral blood stem cell transplantation (PBSCT) in patients with poor-risk hematological malignancies and solid tumors. A single centre experience. AB - Between July 1994 and December 1996, PBSC were mobilized in 28 patients with poor risk hematological malignancies and solid tumors. CD34+ cells were positively immunoselected using the Ceprate CS System. By December 1996, 22 patients had been reinfused with a median of 3.325 (0.078-9.5) x 10(6)/kg CD34+ cells. In three patients unselected back-up PBSC had to be transfused along with selected CD34+ cells because of a CD34+ cell number <0.5 x 10(6)/kg. G-CSF (10 microg/kg) was started on day +1 and all patients engrafted within a median day number of 12 (range, 10-22) until leukocytes >1.0 x 10(9)/l and a median day number of 56 (range, 10-180) until platelets >20.0 x 10(9)/l (ie platelet transfusion independence). Time to leukocyte and platelet recovery was significantly shorter in patients receiving >2.0 x 10(6)/kg purified CD34+ cells as compared to patients reinfused with <2.0 x 10(6)/kg CD34+ cells. The hematopoietic recovery time was similar to that of 18 historical control patients treated with unseparated ABMT +/- PBSCT with the exception of a significantly faster leukocyte engraftment in patients receiving >2.0 x 10(6)/kg CD34+ cells and a significantly delayed platelet recovery time in patients receiving <2.0 x 10(6)/kg purified CD34+ cells. There was a trend for a better overall survival and a lower probability of progression/relapse as compared to the historical controls. We observed five episodes of serious opportunistic infections (three pulmonary fungal infections, two cases of cryptosporidiosis) after the take. Four of these patients had been reinfused with <2.0 x 10(6)/kg CD34+ cells probably indicating a delayed immune reconstitution after CD34+-selected PBSCT. PMID- 9404925 TI - A phase I-II study of high-dose melphalan, mitoxantrone and carboplatin with peripheral blood stem cell support in patients with advanced ovarian or breast carcinoma. AB - The purpose of this study was to develop a high-dose chemotherapy (HDC) and peripheral blood stem cell (PBSC) regimen for treatment of patients with ovarian carcinoma that could be administered in an outpatient setting. Fourteen patients with advanced ovarian (n = 9) or breast (n = 5) carcinoma, who had failed conventional chemotherapy, were entered into a dose-escalation trial to determine the maximum tolerated dose (MTD) of carboplatin that could be administered with fixed doses of melphalan (160 mg/m2) and mitoxantrone (50 mg/m2). Twenty-five additional patients were included in a phase II trial at the MTD. Two of two patients had grade 4 severe regimen-related toxicities (RRT), one fatal, at a dose level of 1600 mg/m2. Two of 29 patients (6.9%) treated at the MTD (carboplatin, 1400 mg/m2) died of RRT. All three patients who died of toxicity had a calculated AUC for carboplatin >30 mg/ml/min. Thirty-one patients with ovarian cancer who had failed chemotherapy were treated, 24 at the MTD. Fourteen of 20 patients (70%) with ovarian carcinoma with evaluable disease achieved a CR and seven (35%) are alive disease-free a median of 20 months (range, 7-26). Five of seven patients with ovarian cancer who had failed chemotherapy but were rendered clinically disease-free following surgery survive without progression a median of 13 months (range, 9-19). Eight of 16 (50%) platinum-resistant and 4/12 (33%) platinum-sensitive patients with ovarian cancer survive disease-free. PMID- 9404927 TI - Allogeneic bone marrow transplantation in patients who relapse after autologous transplantation. AB - Increasing numbers of patients have received autologous stem cell transplants (ASCT) for hematologic malignancies. Since only a fraction of these patients are cured, physicians are more frequently faced with the dilemma of how to manage relapse post-transplant. Potential advantages of allogeneic transplantation (alloBMT) over ASCT include lack of graft tumor contamination and presence of a graft-versus-tumor effect. For this reason, patients who relapse after ASCT are often considered candidates for allogeneic bone marrow transplantation. However, there is limited knowledge on the outcome of alloBMT in patients who relapse after ASCT. We retrospectively analyzed the outcome of 20 patients with malignant lymphoma (n = 14) and AML (n = 6) who underwent alloBMT after failing an ASCT. The median age was 30 (17-41) years and the interval from ASCT to alloBMT was 10.5 (2-25) months. Seventeen patients died between 0.3 to 11 months (median 2.0) after alloBMT, all due to BMT-related toxicities. Three patients remain alive and free of disease at 1.1, 1.2 and 2.5 years after alloBMT. Sixteen of the 18 evaluable patients (89%) developed grade II-IV acute GVHD. Patients undergoing alloBMT after ASCT have a very high treatment-related mortality and incidence of grade II-IV acute GVHD. Alternative treatments with salvage chemotherapy, radiation or investigational approaches should be considered in patients who relapse after ASCT. PMID- 9404926 TI - Efficient peripheral blood stem cell mobilization with low-dose G-CSF (50 microg/m2) after salvage chemotherapy for lymphoma. AB - We compared the use of low-dose G-CSF (50 microg/m2/day), following salvage chemotherapy, for mobilization of PBSC with the results obtained in a comparable historical control group who received a standard dose of G-CSF (5 microg/kg/day, approximately 200 microg/m2/day). Thirty adult patients with relapsed or refractory lymphoma were treated with ifosfamide, VP-16, intermediate-dose Ara-C, methylprednisolone (IAPVP-16) and G-CSF 5 microg/kg/day (group A, n = 15) or 50 microg/m2/day (group B, n = 15) from day 6 until the end of leukaphereses. The duration of neutropenia and thrombocytopenia were equal in both groups. A median of two (1-3) leukaphereses were performed in both groups to harvest >3.5 x 10(6)/kg CD34+ cells. The numbers of circulating CD34+ cells on the first day of leukocyte recovery were similar in both groups in those patients mobilized after a first cycle of IAPVP-16. The numbers of circulating CD34+ cells were similar in patients mobilized after a first and after a second IAPVP-16 in group A. In the low-dose group (group B), however, the numbers of circulating CD34+ cells were significantly lower in those mobilized after a second than after a first course. Additionally, the product of the first leukapheresis contained significantly fewer CD34+ cells in those mobilized after a second course only in group B, with no differences in group A. Nevertheless, the final products harvested did not differ in the content of MNC, CFU-GM and CD34+ cells, suggesting that these differences are not clinically important. These results indicate that the use of low-dose G-CSF (50 microg/m2/day) is as effective as 5 microg/kg/day in accelerating neutrophil recovery and mobilizing CD34+ cells after a first cycle of IAPVP-16 salvage chemotherapy, resulting in a substantial decrease in costs, while more heavily pretreated patients may require higher doses of G-CSF for an equivalent mobilization. PMID- 9404928 TI - Long-term effects of bone marrow transplantation on dental status in children with leukaemia. AB - Minimal data about oral and dental health in long-term survivors after BMT are available. We studied the dental status of 27 children (19 males, eight females) with leukaemia, followed up with a routine oral examination, panoramic tomogram and, when necessary, an endoral radiograph at a median of 2 years (range 1-10) after BMT. Community periodontal index treatment necessity (CPITN), dental caries, missing or filled permanent teeth (DMFT) and dento-facial alterations according to WHO criteria were registered and evaluated. Median age of the patients at BMT was 9 years (range 1.1-17.9). The mean DMFT score ranged from 1.6 to 12.4 according to age at examination and was slightly higher than that which we previously reported in children who received chemotherapy alone. CPITN showed the presence of soft deposits in 77.7%, serious gingivitis in 59.2% and parodontal involvement in 3.7% of cases. Dento-facial abnormalities were found in 55.5% of patients, while 62.9% of the patients had tooth abnormalities or agenesis. Nine out of 27 patients (33%) had root hypoplasia. A negative impact on DMFT index due to multiple post-BMT factors was found. Age is the crucial factor in determining a developmental defect of enamel and root. The follow-up of long term survivors after BMT should include regular dental examination. PMID- 9404930 TI - Dapsone for Pneumocystis carinii prophylaxis in children undergoing bone marrow transplantation. AB - Children who undergo bone marrow transplantation (BMT) are at risk for Pneumocystis carinii pneumonia (PCP). Prophylaxis using trimethoprim/sulfamethoxazole (TMP/SMX) is highly effective but the incidence of adverse drug reactions is significant. We retrospectively reviewed 33 pediatric BMT (25 allogeneic and eight autologous) in whom dapsone was used for PCP prophylaxis because patients were unable to receive TMP/SMX. Dapsone was administered at 50 mg/m2 p.o. once a week from engraftment to 180 days post autologous BMT, and to 1 year or throughout the duration of immunosuppressive treatment post-allogeneic BMT. With a total of 7268 patient days of dapsone prophylaxis and a median follow-up of 353 days post-BMT, no proven PCP was diagnosed. Sixteen cases of chest radiograph abnormalities were noted in this patient population but none was attributed to PCP. Dapsone was well tolerated by all children with no serious adverse effects; however, one patient developed Toxoplasma gondii encephalitis during dapsone prophylaxis. Dapsone warrants further evaluation as an alternative for PCP prophylaxis in pediatric BMT patients intolerant of TMP/SMX. Additional prophylaxis should be considered for patients at high risk for T. gondii encephalitis. PMID- 9404929 TI - Antithrombin-III for the treatment of chemotherapy-induced organ dysfunction following bone marrow transplantation. AB - A hypercoaguable state has been shown to follow high-dose chemotherapy for bone marrow transplantation (BMT). Deficiency of the natural anticoagulants, antithrombin III (AT-III), protein C and protein S correlate with organ dysfunction following BMT. We treated 10 patients with severe post-BMT organ dysfunction with AT-III concentrate. Indications for treatment included AT-III anticoagulant level less than 88% and life-threatening single or multiorgan dysfunction. All patients were loaded with 50 units/kg AT-III every 8 h for three doses followed by 50 units/kg/day each day for 3-12 days. Clinical improvement was seen within 1-5 days of start of therapy in all patients. Patients with veno occlusive disease (VOD) showed a decrease in platelet consumption in nine of nine patients, resolution of hepatic tenderness in six of eight patients, and reduction of severe ascites and weight gain in four of five patients. The probability of death due to VOD and life-threatening organ dysfunction was significantly less in the AT-III-treated group when compared to a historical control group receiving the same preparative regimen (P = 0.047 and P = 0.034, respectively). Significant improvements in organ dysfunction following AT-III treatment in this small study supports a causal relationship between AT-III deficiency and post-BMT chemotherapy-induced organ dysfunction. PMID- 9404931 TI - Difference in the expression of Fas/Fas-ligand and the lymphocyte subset reconstitution according to the occurrence of acute GVHD. AB - Acute graft-versus-host disease (aGVHD) remains a major barrier to a wider application of allogeneic bone marrow transplantation (BMT). Although this complication is mainly dependent on donor-derived T lymphocytes, very little information is available concerning the mechanism of lethality. In this study, we investigated both the expression of Fas/Fas-ligand (FasL) and lymphocyte subset reconstitution in patients who underwent HLA-matched related allogeneic BMT (n = 16) and normal donors (n = 10), and several distinct features were observed. First, the reconstitutions of CD3+ and CD56+ cells were different between the aGVHD+ and aGVHD- group. In particular, the percentage of CD3-CD56+ cells was significantly decreased in patients with aGVHD (P < 0.01). Second, the expansion of CD8+ (P = 0.01) and CD8+ CD28- T cells (P = 0.03) was a characteristic finding in patients with aGVHD. Finally, we found that the percentages of Fas+CD8+, Fas+HLA-DR+ and FasL+ CD8+ cells were significantly increased. Fas antigen was highly coexpressed on most of the lymphocyte subsets, especially on CD8+ cells (P < 0.01), and also, significantly higher coexpression of FasL on CD8+ cells was found in patients with aGVHD (P < 0.01). In summary, an increase in the percentage of CD8+ cells which express Fas and its ligand in patients with aGVHD after BMT points to a possible role for the Fas/FasL pathway in the effector phase of aGVHD. PMID- 9404932 TI - Cost analysis of the introduction of PBPC for autologous transplantation: effect of switching from bone marrow (BM) to peripheral blood progenitor cells (PBPC). AB - Increasingly, PBPC instead of BM are used for autologous transplantation. Limited data exist on the economic effects of this change. Using a resource-based utilization model we prospectively determined the costs of 48 autologous transplants (eight BM, 17 BM + PBPC, 23 PBPC), isolating the post-reinfusion period (day 0 to discharge) to better determine the effect of the rescue product. Length of stay post-reinfusion was significantly shorter in patients receiving PBPC (median 13 days) or BM + PBPC (median 14 days) vs BM alone (median 20 days) (P < 0.01). Accordingly, transplant admission costs were less in the PBPC groups (PBPC $22089, BM + PBPC $23179) vs the BM alone group ($32289) (P < 0.05). Rescue product acquisition costs were higher for PBPC (range $3439-$5157) vs BM ($2766) but these costs were offset by the more rapid recovery of patients receiving PBPC. Overall transplant costs depend on the conditioning regimen with a 10-fold cost variation among regimens. Modeled costs for autologus transplantation using various approaches to rescue product acquisition are given. The introduction of PBPC for autologus transplantation has resulted in cost savings at our institution. Although the acquisition costs of PBPC rescue product are greater than for BM, this incremental expense is more than offset by a less expensive post-reinfusion period. PMID- 9404933 TI - Hepatic injury localized to the field of total lymphoid irradiation. AB - A 37-year-old woman with severe aplastic anemia underwent allogeneic bone marrow transplantation following cyclophosphamide (CY) and total lymphoid irradiation (TLI). On day +30, a CT scan was carried out because of a mild elevation in liver enzymes, and it revealed a low density area with a sharp border in the left lobe corresponding to the irradiated area. MRI showed a hypersignal intensity on both T1 and T2-weighted images and suggested that hepatic damage was mainly severe fatty change. These abnormalities resolved with no treatment. CY with TLI for adult patients with severe aplastic anemia may induce hepatic injury. PMID- 9404934 TI - Consolidation therapy with autologous stem cell transplantation in plasma cell leukemia after VAD, high-dose cyclophosphamide and EDAP courses: a report of three cases and a review of the literature. AB - Plasma cell leukemia (PCL) is a rare lymphoproliferative disorder characterized by a malignant proliferation of plasma cells in blood and bone marrow. Treatment of primary PCL has been mostly disappointing. Three patients with primary PCL are described who received high-dose melphalan with autologous PBSC support after vincristine, doxorubicine and dexamethasone (VAD), high-dose cyclophosphamide, and etoposide, cisplatinum, dexamethasone and cytosine arabinoside (EDAP) courses. All patients were in CR post-transplantation. One patient relapsed after 3 months; the other patients are still in CR, after 14 and 26 months, respectively. These results in conjunction with data from the literature suggest that intensive chemotherapy for PCL is promising. PMID- 9404935 TI - Human herpes virus-6 encephalitis after bone marrow transplantation: successful treatment with ganciclovir. AB - Here we report a case that presented with sudden onset of neurological symptoms and was treated with ganciclovir. Polymerase chain reaction analysis for human herpes virus-6 (HHV-6) from his cerebrospinal fluid was later found to be positive. He subsequently recovered with minimal residual neurological symptoms. Encephalitis secondary to this virus may be more common than previously thought in patients undergoing bone marrow transplantation. This condition should be considered in the differential diagnosis of sudden onset neurological symptoms after bone marrow transplantation. PMID- 9404936 TI - Allogeneic bone marrow transplantation for AL amyloidosis. AB - AL amyloidosis is an infiltrative disorder characterized by the extracellular deposition of insoluble fibrillar immunoglobulin light chains whose production results from a plasma cell dyscrasia. Treatment with melphalan has resulted in an improvement in a few patients. Recently, intensive chemotherapy followed by autologous or syngeneic stem cell support has been shown to offer potential benefit. Allogeneic stem cell support after intensive therapy would retain the benefits of autologous transplantation, with the additional advantages of a tumor free graft and of a possible graft-versus-tumor effect. We report a patient with AL amyloidosis and significant proteinuria. She improved after an allogeneic bone marrow transplantation. PMID- 9404937 TI - NMDAR1-like immunoreactive fibers appear in the ipsilateral optic tract during optic nerve regeneration in Rana pipiens. AB - N-Methyl-D-aspartate receptor subunit 1-like immunoreactivity (NMDAR1-LI) was investigated in the brain of Rana pipiens during optic nerve regeneration. Following unilateral optic-nerve crush, frogs were tested for prey-catching and optokinetic nystagmus responses to assess return of visual function. At 1, 2, 3 and 5 months after the surgery, NMDAR1-LI was assessed in central visual pathways. At 3 and 5 months, conspicuous ipsilateral NMDAR1-LI fibers were detected in the thalamic and pretectal nuclei, and the time of their appearance coincided with the onset of behavioral recovery. Also, only ipsilateral retinorecipient layers in the optic tectum showed increased NMDAR1-LI during optic nerve regeneration. These results suggest that NMDA receptors may be present on retinal ganglion cell axons and terminals that have been misrouted during regeneration. PMID- 9404939 TI - Modification of group I field potentials in the intermediate nucleus of the cat spinal cord after chronic axotomy of an extensor nerve. AB - In walking decerebrate cats the influence of group I afferents from the medial gastrocnemius (MG) and the lateral gastrocnemius/soleus (LGS) muscles on stance phase duration is altered after axotomy of the LGS nerve [Whelan, P.J., Hiebert, G.W. and Pearson, K.G., J. Neurophysiol., 74 (1995) 2782-2787]. We examined whether a site for this plasticity is the connection from group I afferents onto spinal interneurons. Group I field potentials from MG, LGS and plantaris (PL) muscle afferents were recorded in the intermediate nucleus of the L6/L7 segments. Within 5 days following the transection of the LGS nerve in one hind leg, the field potential amplitudes from LGS afferents were decreased, from MG increased and those from PL were unchanged. The changes in the field potentials parallel modifications in the influence of group I afferents from the MG and LGS muscles on stance phase duration during walking. PMID- 9404938 TI - Modulation of c-fos expression in the rat striatum by diazepam. AB - The benzodiazepine agonist, diazepam, inhibits cAMP production in the rat brain. Since cAMP influences c-fos activity, we examined the effects of diazepam on expression of this immediate early gene, as indicated by Western blot analysis. Intraperitoneal administration of diazepam increased Fos protein levels in the striatum, but not in the hippocampus. In contrast, pretreatment with diazepam blocked the potent inducing effect of amphetamine, in both brain regions. Similar induction and blockade effects were also observed for a 90 kDa Fos related antigen (Fra), in the striatum and hippocampus. The possible mechanisms underlying the modulatory effects of diazepam on c-fos expression in the brain are discussed. PMID- 9404940 TI - Oxidative metabolism in cultured fibroblasts derived from sporadic Alzheimer's disease (AD) patients. AB - Fibroblasts from Alzheimer's disease (AD) patients displayed decreased cytochrome c oxidase (complex IV) activity (P < 0.05). The basal oxygen consumption rate (QO2) and the response to an uncoupler of oxidative phosphorylation did not differ between AD and control fibroblasts. The QO2 of AD fibroblasts was more susceptible (P < 0.05) to inhibition by azide in the range 0.5-5 mM. The basal intracellular pH (pHi) in AD fibroblasts was significantly more acidic than in control ones. The results support the hypothesis that subtle dysfunctions of oxidative energy-producing processes are present in fibroblasts from sporadic AD patients. The alterations observed scantly influence the fibroblasts functioning even in stressful conditions; however in tissues, such as the brain, that rely heavily on oxidative metabolism for their function, similar alterations may trigger molecular mechanisms leading to cell damage. PMID- 9404941 TI - Protein kinase C in the parabrachial nucleus of rats during conditioned taste aversion induced by amphetamine. AB - D-Amphetamine (AM) is a potent inducer of conditioned taste aversion (CTA) the mechanism of which differs from that induced by lithium. The aim of the present communication is to see whether AM-induced CTA will produce shift in the protein kinase (PKC) activity in the parabrachial nucleus (PBN). Activity of PKC was measured in PBN of rats during AM-induced CTA. In the control experiments a single intraperitoneal (i.p.) injection of AM (3 mg/kg) alone (not paired with saccharin drinking) resulted in rise of particulate bound PKC by 77% and a tendency to decrease its activity in cytosol 60 min but not 24 and 48 h after AM administration. The results suggest translocation of the enzyme from cytosol to membrane. Cytosolic PKC increased by 17 and 50%, 24 and 48 h, respectively, after acquisition of CTA (15 min after the retrieval test), when the direct effect of AM on PKC had already disappeared. Particulate PKC did not change at either of the two time intervals. Thus the total PKC activity was increased. Since we have previously observed the same PKC shifts using LiCl or CuSO4 as CTA unconditioned stimuli, we assume that any CTA inducer will elicit the same alteration of PKC in PBN. PMID- 9404942 TI - Reduced size of right hippocampus in 39- to 80-year-old normal subjects carrying the apolipoprotein E epsilon4 allele. AB - Hippocampal size on magnetic resonance imaging was compared between normal subjects with the apolipoprotein E (apo E) epsilon4 allele (epsilon4/4, epsilon4/3, and epsilon4/2) and those without the epsilon4 allele (epsilon3/3, epsilon3/2, and epsilon2/2) in the age range of 39-80 years. The Mini-Mental State Examination (MMSE) scores did not differ between the two groups. The right hippocampal area and its ratio to hemisphere area and intracranial cavity area were significantly smaller in epsilon4 carriers than non-carriers, whereas hemisphere area did not differ between the two groups. These results suggest that as early as their forties, apo E epsilon4 allele carriers have a markedly smaller right hippocampus with no apparent cognitive impairment, which may have some significance in the high prevalence of the epsilon4 allele in Alzheimer's disease as well as other conditions that cause dementia. PMID- 9404943 TI - Estrogen receptor-immunoreactive neurons in the lumbosacral cord projecting to the periaqueductal gray in the ovariectomized female cat. AB - The periaqueductal gray (PAG) plays a crucial role in reproductive behavior. The present study investigates whether lumbosacral PAG-projecting neurons contain estrogen receptors. In four ovariectomized adult female cats, injections with cholera toxin subunit (CTb) were made into the PAG to retrogradely label PAG projecting neurons in the lumbosacral cord. Estrogen receptor immunoreactive ER IR neurons in the lumbosacral cord were identified immunohistochemically using the antibody H222. PAG-projecting neurons that were immunoreactive for the estrogen receptor were very scarce, and predominantly present in the medial part of the ventral horn. The results indicate that only very few of the neurons relaying information from the urogenital organs to the PAG contain estrogen receptors. PMID- 9404944 TI - Postnatal development of the autonomic and sensory innervation of the musculature in the rat urinary bladder. AB - The postnatal development of the innervation of the muscle layer in the rat urinary bladder was analysed in whole mount preparations using immunohistochemistry against protein gene-product 9.5 (PGP; general neuronal marker), growth-associated protein 43 (GAP), dopamine beta-hydroxylase (DBH), neuropeptide Y (NPY), vasoactive intestinal polypeptide (VIP), calcitonin gene related peptide (CGRP) and substance P (SP). Immunoreactive nerve fibres for all markers were already present at birth. The density of PGP- and GAP-positive nerve fibres was similar and remained constant throughout the postnatal development. The rank order of densities for the other markers relative to PGP was NPY (129 189%) > CGRP (20-63%) > SP (7-23%) > DBH (7-12%) > VIP (2-11%). While the density of presumably efferent VIP- and DBH-positive fibres did not change postnatally, NPY-positive fibres reached adult density at the fifth postnatal day. Sensory CGRP- and SP-positive nerve fibres approached adult levels at the end of the second week, shortly before the micturition reflex was completely developed. The data suggest that a sufficient relative density of sensory and certain efferent elements might be a prerequisite for the development of the mature micturition reflex. PMID- 9404945 TI - Acute sodium deficiency reduces gustatory responsiveness to NaCl in the parabrachial nucleus of rats. AB - Sodium-deprived rats ingest excessive amounts of salt solutions at high concentrations which are normally avoided. This phenomenon, salt appetite, is known to be controlled by the sense of taste. In the present study, a possible involvement of the parabrachial nucleus (PBN) in sodium appetite was examined by recording PBN neuron responses to 12 different taste stimuli in sodium-deprived and control groups of urethane anesthetized rats. Sodium deficiency was induced by two injections of furosemide 24 and 22 h before the recording. Taste responses to 0.3 and 0.5 M solutions of NaCl were significantly lower in the sodium deprived rats than in controls. In gustatory responses to other taste solutions except for quinine hydrochloride solution, no differences were detected between the two groups of the rats. Correlation coefficients of responses among sodium salts and those between 0.5 M NaCl and sweeteners were larger in the sodium deprived rats than in controls. The results indicate that PBN neurons in sodium deprived rats are critically implicated in induction of quantitative and qualitative changes of gustatory sense to sodium salts. PMID- 9404946 TI - Actions of barium on rapidly inactivating potassium current in bullfrog sympathetic neurons. AB - Whole-cell/voltage-clamp recordings were made from dissociated bullfrog sympathetic neurons to examine the inhibitory actions of barium (0.01-3 mM) on a rapidly inactivating A-type potassium current (IA). The IC50 value was about 0.9 mM. Barium (1 mM) approximately halved the maximum amplitude of IA (approximately 1.7 nA near 0 mV) without significantly affecting a voltage for the 50% activation (approximately -40 mV) and that for the 50%-inactivation (approximately -90 mV), nor did it affected the time course of IA. The results suggest that the barium block is independent of the kinetics of the A-channels in bullfrog sympathetic neurons. PMID- 9404947 TI - Neurotrophin-3 mRNA expression in rat intrafusal muscle fibres after denervation and reinnervation. AB - We have studied the regulation of the expression of neurotrophin-3 (NT-3) mRNA in neonatal and adult rat muscle spindles after denervation and after denervation followed by reinnervation. Denervation of the intrafusal fibres did not result in an upregulation of the NT-3 mRNA expression but decreased this expression below the detection limit of the in situ hybridization method. Reinnervation of intrafusal fibres restored the NT-3 mRNA expression. The results suggest that the expression of NT-3 mRNA in postnatal muscle spindles is controlled by neuronal factors. The intrafusal fibre derived NT-3 may act as an instructive, feedback messenger for innervating neurons and may play a role in stabilizing and maintaining functional neuromuscular connections. PMID- 9404948 TI - U-50,488H, a selective kappa opioid receptor agonist, ameliorates memory impairments induced by muscarinic autoreceptor agonist, carbachol in mice. AB - We investigated whether carbachol, a muscarinic receptor agonist, induces learning and memory impairments, and U-50,488H, a selective kappa opioid receptor agonist, ameliorates the impairments of learning and memory using a step-down type passive avoidance task in mice. Carbachol induced a dose-related dual response. Carbachol (3 nmol/mouse, i.c.v.) significantly shortened the step-down latency, while lower (1 nmol) and higher (10 nmol) doses of carbachol did not induce learning and memory impairments. U-50,488H (0.64 micromol/kg, s.c.) significantly improved carbachol-induced impairments of learning and memory. These findings suggest that kappa opioid receptor agonists ameliorate learning and memory impairments which may associate with dysfunction of presynaptic cholinergic neurons. PMID- 9404949 TI - Deficiency of selenium enhances the K+-induced release of dopamine in the striatum of mice. AB - To determine whether a selenium (Se) deficiency in the brain leads to a functional change in dopaminergic transmission in the striatum, in vivo microdialysis was conducted in mice fed a low-Se diet. After 11-13 weeks of the diet regimen, the activity of glutathione peroxidase (GPx) in the Se-deficient brain was reduced to 60% of the control brain. A high K+ perfusion (100 mM) increased the level of dopamine in the dialysate to 67 +/- 16 times the basal level; the increase was significantly greater than that observed in the control group (28 +/- 4 times). Such a between-group difference was not observed after 4 5 weeks of the Se-diet. These results indicated that prolonged Se deficiency altered the function of striatal dopaminergic neurons in mice. A possible contribution of enhanced oxidative stress due to the reduced GPx activity is discussed. PMID- 9404950 TI - Mechanisms and sites of pyrogenic action exerted by staphylococcal enterotoxin A in rabbits. AB - The febrile responses induced by i.v. administrations of staphylococcal enterotoxin A (SEA) was mimicked by direct injection of SEA into the organum vasculosum laminae terminalis (OVLT) in unanesthetized rabbits. Compared with the febrile responses induced by i.v. injection of SEA, the OVLT route of injection required a much lower dose of SEA to produce a similar fever. Furthermore, the fever induced by intra-OVLT or i.v. injection of SEA was significantly attenuated by pretreatment with intra-OVLT injection of anisomycin (a protein synthesis inhibitor), indomethacin or diclofenac (inhibitors of cyclo-oxygenase (COX)), and aminoguanidine or dexamethasone (inhibitors of inducible nitric oxide synthase (iNOS)). These results suggest that COX or iNOS pathway in the OVLT mediate the SEA-induced fever in rabbits. PMID- 9404951 TI - Dihydrexidine produces hypothermia in rats via activation of dopamine D1 receptors. AB - The selective dopamine D1 receptor agonist dihydrexidine (2.0-8.0 mg/kg, s.c.) caused a dose-dependent decrease in core temperature in rats. The hypothermia produced by dihydrexidine (4.0 mg/kg), was completely blocked by the dopamine D1 receptor antagonists SCH 23390 (0.1 mg/kg, s.c.) or NNC 687 (4.0 mg/kg, s.c.), but not by the dopamine D2/3 receptor antagonist raclopride (0.2 mg/kg, s.c.). Neither of the dopamine antagonists by themselves produced any effects on core temperature. The present results provide important evidence for the notion that activation of dopamine D1 receptors induces hypothermia in rats. PMID- 9404952 TI - Peripheral non-opioid analgesic effects of kyotorphin in mice. AB - Bradykinin (BK) given into the plantar (i.pl.) of the mouse hind-limb produced a flexor response. The flexor responses were dependent on BK doses (0.02-20 pmol, i.pl.), and were completely abolished by Hoe140, a B2-type BK receptor antagonist. Kyotorphin, an analgesic neuropeptide which shows enkephalin release in brain slices, produced a dose-dependent reduction of the BK-induced nociceptive responses in ranges of 10 pmol to 1 nmol (i.pl.). Such analgesic effects of kyotorphin were reversed by leucine-arginine, a specific kyotorphin receptor antagonist, but not by naloxone. The kyotorphin-analgesia was also abolished by pertussis toxin (PTX) pretreatment. These results suggest that peripheral analgesic effects of kyotorphin are mediated through mechanisms of kyotorphin specific receptor and PTX-sensitive Gi/Go, and that the enkephalin release is not necessary for this analgesia. PMID- 9404953 TI - Pathological changes in yearly protocol liver biopsy specimens from healthy pediatric liver recipients. AB - Many centers perform biopsies on transplanted livers annually to assess allograft function because serum biochemical tests do not always correlate with histological findings. Although criteria exist for diagnosing acute cellular rejection, no similar criteria exist to describe the histopathological changes observed in the "normal" liver of an immunosuppressed but healthy child. The purpose of this study was to define the histopathological changes in the allografted livers of healthy children who have undergone transplantation and to evaluate them during long-term follow-up. One hundred fifty-eight yearly protocol liver biopsy specimens of 54 children who received transplants between January 1988 and March 1996 and at least 1 year of follow-up were reviewed, and the biopsy findings were correlated with those of serum tests of liver function performed concomitantly. Thirty-three biopsy specimens were excluded because serum transaminase levels were abnormal, the biopsy specimen was abnormal and diagnostic for a specific lesion, or follow-up showed progression of a specific disease process. In addition, time zero biopsy specimens from 21 of the 54 children were available for comparison. In the protocol biopsy specimens, portal and/or parenchymal mononuclear inflammatory infiltrates were frequent findings (48% and 25%, respectively). Other less common features were mild fibrosis (8%) and focal pericholangitis (6%). Findings in both protocol and time zero biopsy specimens included minimal to mild bile ductular proliferation (15% and 9.5%, respectively) and rare hepatocyte necrosis (1.6% and 5%). No yearly protocol biopsy specimens resulted in any patient benefit. Transplanted livers in immunosuppressed children who are clinically healthy and have normal transaminase levels commonly show histological changes consisting of scattered, mild to moderate, portal and/or parenchymal mononuclear infiltrates that are clinically insignificant. Yearly protocol biopsies in healthy pediatric recipients have been abandoned by the investigators after 3 years of follow-up. PMID- 9404954 TI - Infectious complications after OKT3 induction in liver transplantation. AB - The present study examines the incidence, risk factors, bacteriology, and mortality of infectious episodes and the role of antimicrobial prophylactic regimens after OKT3 induction in liver transplantation. Infections occurring in the first 6 months were evaluated according to the Centers for Disease Control criteria in 102 transplant recipients. Patients were administered OKT3 for 5 to 10 days, beginning intraoperatively, azathioprine, low-dose prednisone, and delayed introduction of cyclosporine. There were 140 major and 30 minor infections for an incidence of 1.7 infections per patient. Twenty-seven patients (26%) had no infectious episodes during the 6 months of follow-up. Bacterial and fungal infections peaked during the first month posttransplantation, whereas viral infections peaked during the second month. Infection-related mortality was 10%. One-year survival rate of patients who suffered a major infection was less than those who were infection free, but the difference was not statistically significant (79% vs. 89%; P = .61). There was a significantly higher incidence of enterococcal infections under cefotetan prophylaxis than under ampicillin sulbactam (.375 vs. 11 infections per patient; P = .0017). There were 14 episodes of cytomegalovirus disease (14%) but no cytomegalovirus-related mortality or graft loss, and all cases responded to ganciclovir treatment. Bivariate and multivariate analyses identified only retransplantation as a risk factor for infection. In conclusion, OKT3 induction after liver transplantation is associated with a manageable incidence of bacterial, viral, or fungal infections. This is caused by, at least in part, improved anti-infective prophylaxis. PMID- 9404955 TI - Neoral in de novo liver transplantation: adequate immunosuppression without intravenous cyclosporine. AB - Absorption of cyclosporine from the traditional oral formulation Sandimmune (Novartis Pharma, Basel, Switzerland) is particularly unpredictable in the early stages after liver transplantation. The absorption of cyclosporine is influenced by liver function, postoperative paralytic ileus, and graft dysfunction. Oral absorption of cyclosporine from Sandimmune is also bile dependent; cholestasis and external biliary drainage are associated with low cyclosporine absorption. Postoperative administration of intravenous Sandimmune is therefore often necessary to obtain adequate immunosuppression, despite the increased risk of renal and neurological toxicity. A microemulsion formulation of cyclosporine, Neoral (Novartis), has been developed to overcome the problems of poor and variable absorption of cyclosporine from Sandimmune. Uptake of cyclosporine from Neoral is rapid and less dependent on bile secretion so that higher peak concentrations are reached and absorption is less variable than with Sandimmune. A review of several open studies in which Neoral was administered to liver transplant patients immediately after transplantation is presented. The results suggest that the use of Neoral as a primary immunosuppressive therapy provides adequate cyclosporine trough levels, minimizing or obviating the need for intravenous cyclosporine administration. In addition, Neoral appears to reduce the risk of acute rejection episodes compared with immunosuppressive regimens involving intravenous cyclosporine. PMID- 9404957 TI - An outbreak of vancomycin-resistant Enterococcus faecium in liver transplant recipients. AB - Vancomycin-resistant Enterococcus faecium (VREF) has become a significant nosocomial pathogen for immunosuppressed patients. During a 5-month period in 1993, 8 cases of invasive infection with VREF (7 with bacteremia) were identified in liver transplant recipients, half of whom were adults. Epidemiology and microbiology studies were designed to identify the source and to determine the risk factors for this infection. Overall mortality was 50% (3 adults and 1 child). Mortality in bacteremic patients was 57%. A case-control study showed that cases were more likely to have been treated with a third-generation cephalosporin or vancomycin and to have undergone more than four biliary tract procedures. Environmental surveillance cultures yielded only one VREF isolate from a rectal temperature probe, but this device was used in only 2 of the cases. Cultures from all surgery and radiology suites were negative. All VREF isolates were genotyped by contour-clamped homogenous electric field electrophoresis of chromosomal DNA restriction fragments. These studies showed that a single clone was responsible for the outbreak, although other clones could be detected in the hospital. After implementing strict contact isolation on the liver transplant unit, only 1 additional patient with VREF was identified during this outbreak. In conclusion, it was found that antibiotic use and biliary tract manipulation were risk factors for developing invasive infections with VREF after liver transplantation. Optimal treatment is still unclear but most likely includes a combination of two or more antibiotics. Prompt institution of infection control measures can preclude rapid spread of this nosocomial pathogen. PMID- 9404956 TI - Hepatitis G virus infection before and after liver transplantation. Liver Transplantation Database. AB - The hepatitis G virus is a newly discovered flavivirus that has been linked to acute and chronic hepatitis of unknown cause. We determined the prevalence of hepatitis G virus infection in 179 selected patients undergoing liver transplantation at three centers participating in the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) Liver Transplantation Database. Pretransplantation and posttransplantation specimens were tested for hepatitis G virus RNA by polymerase chain reaction. Before transplantation, 9 of 38 (24%) patients with fulminant hepatic failure, 9 of 62 (15%) with cryptogenic cirrhosis, 3 of 35 (9%) with cholestatic liver disease, and 5 of 44 (11%) with chronic hepatitis C were positive for hepatitis G virus RNA (P = .27). Patients with and without viral RNA were similar in clinical features, liver test abnormalities, and survival after transplantation. Posttransplantation serum specimens were tested from 73 patients; 9 of 11 (82%) who were positive for viral RNA before transplantation remained positive, but 35 of 62 (56%) patients who were initially negative became positive after transplantation, a rate consistent with that predicted from the number of blood products administered. Only 5% of de novo HGV infections could be attributed to preexisting hepatitis G virus RNA in the donor. Comparison of patients with and without hepatitis G virus infection showed no difference in incidence of hepatitis after transplantation. Thus, hepatitis G virus infection was present in 15% of patients before and appeared de novo in half of patients after liver transplantation. Although hepatitis G virus infection was not associated with poor outcome, the frequency of this infection after transplantation calls for further long-term evaluation. PMID- 9404958 TI - Posttransplant eosinophilic gastroenteritis in children. AB - The cause of eosinophilic gastroenteropathy in older children and adults is unknown. In this report, two post-liver transplantation children treated with low dose cyclosporine A and alternate-day low-dose prednisone are described who were administered a single bolus administration of a lympholytic dose of corticosteroids without taper and who developed intestinal symptomatology several weeks later. Histologic examination of mucosal biopsy specimens from various regions of the gastrointestinal tract showed an intense eosinophilic infiltration of the mucosa and lamina propria. The patients recovered after corticosteroid administration was tapered. Post-transplant gastroenteric eosinophilic inflammation may need to be considered in patients on immunomodulatory medications who have chronic intestinal symptomatology. PMID- 9404959 TI - Hemodynamic effects of inhaled nitric oxide in four patients with severe liver disease and pulmonary hypertension. AB - Patients with moderate and severe pulmonary hypertension have a very high mortality rate when undergoing orthotopic liver transplantation. Because nitric oxide has been successful in reducing pulmonary artery pressures in certain patients with pulmonary hypertension, the efficacy of NO inhalation (40 and 80 ppm) in 4 patients with pulmonary hypertension associated with liver disease was determined. No clinically significant changes in pulmonary artery pressures or other hemodynamic parameters were observed using either concentration of NO. In conclusion, no pulmonary vasodilatory response from inhalation of NO in 4 patients with severe liver disease and pulmonary hypertension was found. PMID- 9404960 TI - Low-dose aspirin therapy is associated with few side effects but does not prevent hepatic artery thrombosis in liver transplant recipients. AB - Hepatic artery thrombosis occurs in 4% to 10% of adult patients and in up to 26% of children undergoing liver transplantation. Aspirin has been used to prevent this complication but may increase procedure-related and gastrointestinal bleeding. The aim of this study was to assess the efficacy and safety of low-dose aspirin in the prophylaxis of hepatic artery thrombosis. The histories of 529 patients who survived liver transplantation between September 1988 and December 1993 were reviewed retrospectively. The routine clinical practice followed until 1992 was to initiate oral aspirin therapy on the first postoperative day (81 mg daily in adults and 40 mg daily in children) as prophylaxis for vascular thrombosis. This was done in 354 patients. Aspirin was not administered to the remaining 175 patients. Hepatic artery thrombosis occurred in 13 patients treated with aspirin (3.7%) and in 7 patients not treated with aspirin (4.0%) (P = .85). Recipient age of younger than 2 years and low donor liver weight were the only factors that predisposed the patients to hepatic artery thrombosis. A total of 1,651 percutaneous liver biopsies were performed in this series, with 1,111 performed in patients treated with aspirin. Significant bleeding after liver biopsy occurred in 12 patients treated with aspirin (1.1%) and in 3 patients not treated with aspirin (0.6%) (P = .29). Gastrointestinal bleeding occurred in 66 patients treated with aspirin (18.9%) and in 23 patients not treated with aspirin (12.8%) (P = .08). Low-dose aspirin therapy is not shown to be effective in preventing hepatic artery thrombosis after liver transplantation. Although aspirin does not produce a statistically significant increase in the risk of bleeding after liver biopsy, there is a trend toward an increased incidence of gastrointestinal bleeding. PMID- 9404961 TI - Hepatic artery resistance index can predict early death in children with biliary atresia. AB - Hepatic artery resistance index has been measured by ultrasonography Doppler and has been found to predict rapid deterioration and death in children with biliary atresia. Clinical, biochemical, ultrasonographic, and outcome data were collected prospectively and retrieved on 32 patients with resistance index of > or = 1.0 (group A). These were compared with the same data for 32 age- and sex-matched patients with biliary atresia and a resistance index of < 1.0 (group B). Group A was found to have significantly worse liver function tests than group B. In group A, all patients died (n = 11) or underwent transplantation (n = 21; of whom 4 died) compared with only 2 patients who died in group B and 4 patients who underwent transplantation without fatality. Survival at 2 years was 52% in group A v 94% in group B. It is suggested that regular ultrasonography Doppler examination in patients with biliary atresia can detect a group with a resistance index of > 1.0 who have a very high risk of early mortality. Such patients require early evaluation and listing for transplantation. Those listed for liver transplantation on other grounds require ultrasonography examinations every 2 to 3 months with immediate upgrading of the priority of those patients found to have a resistance index of > or = 1.0. PMID- 9404962 TI - Gastric mucosal pH is associated with initial graft function but is not a predictor of major morbidity after liver transplantation. AB - Gastric mucosal pH reflects splanchnic perfusion. Monitoring gastric mucosal pH might be useful in predicting outcome after liver transplantation. Forty patients were included in the study. Gastric mucosal pH and gastric mucosal pH corrected for systemic pH were compared with regard to initial liver function and morbidity. Eighty percent of the patients had at least one episode with a gastric mucosal pH of <7.32, and 84% of these had a concomitant arterial pH of <7.32. No differences in morbidity were found between patients with a gastric mucosal pH of <7.32 and those with a gastric mucosal pH of >7.32. If gastric mucosal pH was corrected for arterial pH, only 49% of the patients had an episode during transplantation with a corrected gastric mucosal pH of <7.32. Comparing these patients with the group that did not have such an episode, we found that flow in the venovenous bypass system was significantly lower (2.9 v 3.4 L/min; P < .02) in the first group. Also alanine aminotransferase and aspartate aminotransferase levels were higher, antithrombin III levels and lidocaine clearance rates were lower, and prothrombin times were longer in the group with corrected gastric mucosal pH of <7.32. No differences with regard to major morbidity and mortality were noted. Gastric mucosal pH during liver transplantation should be corrected for arterial pH. Patients with a corrected gastric mucosal pH of <7.32 are more likely to develop initial liver function tests disturbances, but morbidity is not different from patients with gastric mucosal pH of >7.32. PMID- 9404963 TI - Vascular occlusion in hepatic resections in cirrhotic rat livers: an experimental study in rats. AB - The aim of this study was to evaluate the tolerance of normothermic liver ischemia with different degrees of hepatic function in cirrhotic rats. Liver cirrhosis was induced by administering carbon tetrachloride (CCl4) in water solution to male Wistar rats. Hepatic function was graded using the plasma levels of antithrombin III, albumin, and bilirubin and the presence of ascites. Rats were distributed in four groups: noncirrhotic (control group), compensated cirrhosis (group A), decompensated cirrhosis (group B), and decompensated cirrhosis with ascites (group C). Groups A, B, and C were significantly different in all four parameters studied (P < .003). Subtotal liver ischemia was performed for periods of 0, 30, 45, 60, and 75 minutes. At the end of the procedure, the nonischemic lobes were resected. Postoperative evolution of alanine aminotransferase, aspartate aminotransferase, and bilirubin levels was also recorded. Survival rates after the same periods of ischemia were statistically different (P < .05): control group, 7 of 7 after 45 minutes (100%), 7 of 7 after 60 minutes (100%), and 4 of 9 after 75 minutes (44%); group A, 7 of 7 after 45 minutes (100%) and 1 of 7 after 60 minutes (14%); group B, 7 of 7 after 0 minutes (100%), 5 of 7 after 30 minutes (71%), and 1 of 7 after 45 minutes (14%); and group C, 0 of 5 after 0 minutes (0%) and 1 of 7 after 30 minutes (14%). No differences were found in the postoperative course of transaminases. However, bilirubin levels found 24 hours and 7 days after ischemia were significantly greater in cirrhotic rats, and this was directly related to the degree of hepatic insufficiency (P < .001). Histological examination of the livers exposed to CCl4 showed features of liver cirrhosis with ductal proliferation. The ischemia time tolerated by cirrhotic rat livers is shorter than the time tolerated by normal rats. Tolerance to hilar vascular occlusion depends on the degree of hepatic insufficiency. Rats with decompensated cirrhosis and ascites do not tolerate any surgical procedure. PMID- 9404964 TI - Rationale and technical constraints of a tertiary liver transplantation. AB - Because of the current shortage of donor organs, the routine performance of tertiary liver transplantation (LT) may be questioned. In this study, the indications of tertiary LT are discussed, paying particular attention to intraoperative technique. Of 501 LTs performed from 1986 to 1995, eight (1.6%) were tertiary LTs. Three patients underwent an emergent third LT because of associated hepatic artery and portal vein thromboses (n = 2) or hyperacute rejection (n = 1). Five patients had an elective third LT for ischemic cholangitis (n = 4) or chronic rejection (n = 1). The 3 patients who underwent retransplantation emergently died early from multiple-organ failure. Because of previous surgery and subsequent technical difficulties, the third LT in the remaining 5 patients required unroutine surgical procedures including the following: intrapericardial control of the suprahepatic vena cava (n = 1), "en bloc" clamping of both the infrahepatic vena cava and the hepatic pedicle (n = 1), arterial reconstruction onto the aorta via an aortoiliac conduit (n = 5), and aortic resection with aortoaortic prosthetic reconstruction (n = 1). Of these 5 patients, 4 required reoperation because of bowel perforation (n = 5) or intraperitoneal bleeding (n = 1). The 5 patients (62%) who were regrafted electively are alive and well after a median follow-up of 45 months. A third LT can be reasonably offered to selected young recipients if performed electively. Tertiary LT may require unroutine surgical procedures and may lead to severe morbidity. PMID- 9404965 TI - Minimal criteria for placement of adults on the liver transplant waiting list: a report of a national conference organized by the American Society of Transplant Physicians and the American Association for the Study of Liver Diseases. AB - This report summarizes a recent meeting cosponsored by the American Society of Transplant Physicians and the American Association for the Study of Liver Diseases to formulate minimal criteria by which patients with severe liver disease will be placed on the waiting list for liver transplantation. The participants agreed that only patients in immediate need of liver transplantation should be placed on the waiting list. Patients should not be placed in anticipation of some future need for such therapy. It was agreed that minimal criteria could assist but not replace the clinical judgment of the transplant professionals at individual centers. The criteria will be summarized below for adult patients with acute or chronic liver disease. The most important non disease-specific criterion for placement on the transplant waiting list was an estimated 90% chance of surviving 1 year. This translated into a Child-Pugh score of > or = 7 for patients with cirrhosis which places the patient in Child-Pugh class B or C. Cirrhotic patients who have experienced gastrointestinal bleeding caused by portal hypertension or a single episode of spontaneous bacterial peritonitis would meet the minimal criteria irrespective of their Child-Pugh score. There were disease-specific criteria also. These include a sole minimal criterion for patients with fulminant hepatic failure regardless of etiology of the onset of stage 2 hepatic encephalopathy. A requirement for 6 months abstinence from alcohol before placement on the transplant waiting list was considered appropriate for most patients with alcoholic liver disease. Exceptional cases could get access to the waiting list through a regional review process. Chronic cholestatic diseases present difficulties because of a different natural history than that of chronic hepatocellular diseases. The use of specific risk scores for primary biliary cirrhosis and primary sclerosing cholangitis will likely replace Childs-Pugh classification as the scoring systems become refined. Minimal criteria for any patient with a primary hepatocellular cancer would admit any patient with a tumor confined to the liver irrespective of size or number of tumors, after careful investigation had failed to show spread to lymph nodes, the portal vein, or distant organs. Unusual or rare indications for liver transplantation, including Budd-Chiari syndrome, Wilson's disease, and other hereditary disorders, were also discussed. Finally, it was agreed that there should be no absolute contraindications to placement of patients on the liver transplant waiting list. These criteria should be open to regular review to accommodate advances in the field. PMID- 9404967 TI - Splenorenal shunt closure after liver transplantation: intraoperative Doppler assessment of portal hemodynamics. PMID- 9404966 TI - Persistent cortical blindness after cyclosporine leukoencephalopathy. AB - Cyclosporine A (CyA)-related cortical blindness is an uncommon complication of CyA therapy in patients undergoing liver transplantation. Characteristically, neurological symptoms associated with CyA treatment usually regress after cyclosporine withdrawal. We present a case of a liver transplant recipient in whom discontinuation of CyA therapy has resulted in only partial clinical improvement, and cortical blindness remains after 1 year of follow-up. The evolution of cerebral magnetic resonance imaging (MRI) findings over time in this form of CyA leukoencephalopathy is also described. PMID- 9404968 TI - Interleukin-2 receptor antibody therapy. PMID- 9404971 TI - A surgeon's perspective on the management of coagulation disorders before liver transplantation. PMID- 9404970 TI - A hepatologist's perspective on the management of coagulation disorders before liver transplantation. PMID- 9404972 TI - Transfusion based on clinical coagulation monitoring does reduce hemorrhage during liver transplantation. PMID- 9404973 TI - Coagulation techniques are not important in directing blood product transfusion during liver transplantation. AB - Preoperative acquired clotting parameters such as prothrombin time, activated partial thromboplastin time, antithrombin III, platelet concentration, and fibrinogen show coagulopathy caused by insufficiency of the diseased liver. Intraoperative determination of clotting factors or parameters is not helpful to direct intraoperative transfusion of blood, blood components, or platelets because transfusions performed solely for correction of clotting data do not correlate with the real intraoperative requirements and increase the costs of orthotopic liver transplantation. However, the use of antihyperfibrinolytic drugs seems to reduce intraoperative blood loss. Patients with cirrhotic disorders caused by portalvenous hypertension show extensive collaterals and increased intravascular blood volume. Thus it is plausible that especially overcorrection of blood loss during the surgical preparation in the preanhepatic phase of the operation results in extensive blood loss. Therefore, to avoid blood loss we attempt to keep volume substitution to a minimum during the preanhepatic phase of the operation. In contrast, during the anhepatic and postanhepatic phases we attempt to reestablish normovolemia by transfusing red packed blood cells and fresh-frozen plasma. Strictly confined use of blood products in the preanhepatic phase, followed by later correction of intravascular blood volume, may reduce intraoperative blood loss; it also seems to ensure adequate substitution of clotting factors. PMID- 9404974 TI - The use of antifibrinolytic agents results in a reduction in transfused blood products during liver transplantation. PMID- 9404975 TI - Antifibrinolytics do not reduce transfusion requirements in patients undergoing orthotopic liver transplantation. PMID- 9404976 TI - Hepatitis G virus: what is the next step? PMID- 9404977 TI - A collective wisdom. PMID- 9404978 TI - Recurrence of nonalcoholic steatohepatitis (NASH) post-liver transplantation. PMID- 9404979 TI - The 1997 Moyer Award. Cytokine production in patients with hypertrophic burn scars. AB - To clarify the significance of the role of the immune system in the formation of proliferative burn scars, this study attempted to identify differential production of cytokines between patients with burn injuries with and without hypertrophic scars. Mononuclear cell fractions were isolated from the peripheral blood (PBMC) of each patient and incubated with and without antigenic or mitogenic stimulation. The resultant supernatants were then assayed by ELISA techniques for production of various cytokines. The production of IL-1, IL-6, TNF alpha, and TGF-beta2 by unstimulated PBMC was elevated significantly in patients with proliferative scar compared to control patients. Production of TGF-beta2 by stimulated PBMC also was elevated significantly in patients with proliferative scar. This study suggests that an increase in the production of TGF-beta and of proinflammatory cytokines by mononuclear cells may play a significant role in the processes that lead to excessive scar formation after burn injury. PMID- 9404980 TI - The 1997 Lindberg Award. Effects of burn injury on bone and growth in a mouse model. AB - Bone growth and remodeling are inhibited by severe burns in adult and pediatric patients, resulting in alterations in linear growth, bone mass, osteoporosis, and increased risk for pathologic fractures. This study of a mouse model of burn injury showed skeletal changes similar to those reported in patients with burn injuries. Baseline, control, sham, and burned mice were injected with fluorescent markers calcein and tetracycline for histomorphometric analysis. Total femur dry and ash weights and total calcium content were significantly lower 10 days after burn injury compared with sham and control animals. There also were decreases in the percentage of fluorochrome-labeled bone surfaces and bone formation rates in the burn-injured mice compared with control and sham mice; however, there were no differences in the mineral apposition rates. This model now provides an opportunity to examine cellular and molecular mechanisms contributing to skeletal pathology in a well-defined burn injury model. PMID- 9404981 TI - The 1997 Clinical Research Award. Health outcome for burn survivors. AB - Little is known concerning health outcome for patients who survive burn injuries, and how their health outcome compares with that of other medical populations. Such information is important given that the current direction of health care policy decision making is toward outcomes-driven decision models. We compared the health status of 91 patients 1 month after severe burn injury with the published reports of the health status of 39 medical comparison samples, and two reports of health status for the general population. Additionally, we collected longitudinal data on a subsample of our surviving patients with burn injuries at 1 year. Our findings suggest that people who survive a severe burn experience a stable and relatively good health status after their injury compared with other medical samples. However, their health status remains worse than that of the general population over time. Further, people who survive a major burn indicate that the areas of vocational and psychosocial functioning are often the most troublesome for them. PMID- 9404982 TI - Acellular allogenic dermis does not hinder initial engraftment in burn wound resurfacing and reconstruction. AB - Donor site morbidity, including pain and potential hypertrophic scarring, increases directly as donor site thickness increases. Autograft hypertrophy increases inversely to the thickness of autograft harvest. Both of these liabilities might be lessened by the ready availability of a functioning, "off the-shelf" dermal substitute. The most immunogenic components of transplanted allograft skin are the cellular elements of the epidermis and dermis. If these components are removed, the remaining noncellular dermal tissue is relatively immunologically inert. We grafted 10 sites with matched control areas in six patients, with limited (< 25%) areas of the body surface appropriate for donor harvest, who required either acute resurfacing or a reconstructive procedure requiring split-thickness autografting. The six children (three girls and three boys) had an average age of 5.2 +/- 0.9 years (range, 2.8 to 10), and an average burn size of 68.7% +/- 6.7% (range, 47% to 85%). The 10 study and control sites were all parts of 10 separate procedures in these massively burned children, nine sites being reconstructive releases, and one excised acute burn. Successful epithelialization was noted at 7 days over 83% +/- 3.4% (range, 60% to 95%) at the cryopreserved acellular human dermis sites and 83.3% +/- 4.3% (range, 60% to 98%) at the control sites (NS, p = 0.96). Duration of follow-up has been 11.9 +/- 3.5 weeks (range, 2 to 29), with no difference in results by Vancouver Scar Scores. We conclude that cryopreserved acellular human dermis will engraft successfully and support engraftment of overlying thin autograft. Initial results relating to the effectiveness of cryopreserved acellular human dermis in optimizing appearance and function are encouraging, but longer follow-up is required before definitive conclusions can be made. PMID- 9404983 TI - Comparison of pain control medication in three age groups of elderly patients. AB - There are no published reports of burn pain management in the elderly population. To assess the range of requirement and use of opioids among elderly patients with burns of different age categories, a retrospective review of 89 consecutive admissions of patients over 55 years of age (January 1995 through July 1996) was conducted. Complete data were available on 44 patients with a burn mean total body surface area of 17.2%. Patient ages ranged from 55 to 92 years. Individuals were divided into three age categories: Group I (55 to 65) n = 20; Group II (66 to 75) n = 14; and Group III (76 to 92) n = 10. Use of commonly prescribed opioids for procedural pain and breakthrough pain were evaluated. We compared the opioid equivalents of medications prescribed versus the actual amount administered. Paired t tests comparing minimum amount of medication ordered with that given revealed Group I patients received significantly more procedural medication than the minimum prescribed (t = 3.88, p = 0.001), and that Group III patients were given significantly less as needed medication than the minimum prescribed (t = 2.58, p < 0.05). PMID- 9404984 TI - Thermal injury functionally alters bone marrow-derived macrophages: a study of monocyte-hepatocyte interactions. AB - Thermal injury quantitatively and qualitatively alters hematopoiesis, including monocyte-macrophage lineage changes, resulting in altered mononuclear cell function. These bone marrow cells (BMCs) ultimately become fixed tissue macrophages (e.g., Kupffer cells). To study the effects of thermal injury on macrophage-hepatocyte interactions, rat BMCs were isolated 24 hours after burn injury, and myelopoiesis was induced by 7-day culture in granulocyte-macrophage colony-stimulating factor. Separate cultures included inflammatory mediators with growth factor function (IL-6 or PGE2). Cultured cells were incubated up to 96 hours with isolated normal hepatocytes (+/- lipopolysaccharide stimulation). The 96-hour exposure to postburn BMCs produced less of the acute phase proteins (APPs), C3 and transferrin, but more cytotoxicity as measured by 1-lactate dehydrogenase release. Sham BMCs cultured with added IL-6 caused higher APP release and minimal cytotoxicity, whereas burn BMCs stimulated lower APP release and retained cytotoxicity. In conclusion, myeloid cells regulate APP synthesis differently after thermal injury and may become more cytotoxic to hepatocytes. PMID- 9404985 TI - Plasma catalase and glutathione levels are decreased in response to inhalation injury. AB - We determined the effect of a severe smoke exposure on plasma oxidant and antioxidant activity. Adult sheep were given a smoke exposure while under anesthesia that produced a carboxyhemoglobin level of 45% +/- 3%. Twelve sheep were studied; six were given smoke alone and volume-resuscitated with sufficient lactated Ringer's solution to maintain baseline hemodynamics. This response was compared with six control sheep during a 6-hour period. The smoke inhalation injury produced a significant increase in plasma hydrogen peroxide and a significant decrease in plasma lipid peroxidation. Circulating lipid peroxidation did not correlate with tissue lipid peroxidation because lung and liver lipid peroxidation were significantly increased. The plasma antioxidants glutathione, catalase, and vitamin E were significantly reduced in response to the injury. Vitamin C remained unchanged from control. Circulatory failure is not a key element in this study, because lactate levels were controlled with volume resuscitation. The degree of smoke inhalation to the airway produced distant organ lipid peroxidation and a decrease in circulating antioxidants--without producing an increase in circulating lipid peroxidation. Maintaining circulating antioxidants may prevent distant organ lipid peroxidation and may be of clinical use in devising treatment strategies for smoke inhalation injury with the availability of antioxidants. PMID- 9404986 TI - Toxic epidermal necrolysis: an analysis of referral patterns and steroid usage. AB - Toxic epidermal necrolysis (TEN) is an exfoliative disorder associated with epidermal slough and systemic toxicity. As of 1986, the literature has advocated early burn center transfer and has rejected the use of steroids. We questioned whether therapy for TEN has changed to reflect these concepts. All cases of TEN referred to our tertiary burn center since 1988 were reviewed. The history was evaluated for steroid usage and timing of burn center transfer. Drug exposures, septic complications, and deaths were noted. Statistics are expressed as mean +/- SD. Fourteen cases of TEN were identified. Transfer was delayed more than 2 days in 10 (72%) instances. Mean delay was 4.4 +/- 2.7 days. Half received steroids. There were three deaths (21%). Pneumonia developed in five patients (36%), urinary tract infections developed in three (21%) patients, seven (50%) patients required intubation, and three (21%) required hemodialysis. No differences in mortality rates or infectious complications were noted in patients who received steroids or who were transferred late. Septic complications occur frequently in TEN. Delay in transfer and initiation of steroids at referring institutions are common. Early burn center referral and avoidance of steroids needs to be reiterated at the level of the referring physician. PMID- 9404987 TI - Pathogenesis of fever in a rat burn model: the role of cytokines and lipopolysaccharide. AB - We investigated the possible causal relationship between interleukin-6 (IL-6) and increased body temperature (T(B)) in a rat burn model. Transmitters for measuring core temperature and estimating activity were implanted in the abdominal cavity. Animals in the burn group were clipped and received full-thickness scald burns to 45% to 55% of the body surface area, and control animals were clipped. T(B) and activity were measured continuously through the tenth postburn day. Carotid lines were placed, and serial blood samples obtained for lipopolysaccharide, IL-6, and tumor necrosis factor-alpha assay. From the third through the tenth postburn day, the burn group had a consistently significantly higher T(B) during light hours than the control group did (average, 0.45 degrees C +/- 10 degrees C, p = 0.0001). Differences in activity during light hours were not significant between the two groups, therefore, do not account for the observed significant difference in T(B). The average IL-6 serum levels were 3.5-fold higher for the burned animals. In this study, burn and control serum levels of IL-6 demonstrated positive correlation with T(B). These data suggest, but do not prove, a causal relationship between IL-6 and fever in the rat burn model, and make it unlikely that circulating systemic lipopolysaccharide is the cause. PMID- 9404988 TI - Lung compliance, airway resistance, and work of breathing in children after inhalation injury. AB - Pathophysiologic changes associated with inhalation injury make mechanical ventilation in children a challenge. Decreased lung compliance and increased airway resistance after inhalation injury may lead to elevated airway pressures and barotrauma. Previous studies have shown significant decreases in the incidence of pneumonia and death in adult patients with inhalation injury treated with high-frequency percussive ventilation (HFPV) as compared with conventional mechanical ventilation (CMV). No studies to date have compared lung compliance, airway resistance, or work of breathing in children being treated with HFPV versus CMV. The purpose of this study was to evaluate lung compliance, airway resistance, and work of breathing in pediatric patients with inhalation injury who required mechanical ventilation. Ten children with bronchoscopically identified inhalation injury requiring mechanical ventilation were studied. Five children received CMV and five children received HFPV. All patients were treated according to our standard inhalation injury protocol. Based on our data and patient population, children receiving ventilation with the HFPV have a significant decrease in the work of breathing as compared with CMV. PMID- 9404989 TI - Acellular human dermis promotes cultured keratinocyte engraftment. AB - In full-thickness skin injury, loss of dermis may result in compromised wound repair, including contracture, hypertrophic scarring, and wound breakdown. This report examines the effect of an acellular dermal matrix on in vivo skin repair. Human keratinocytes cultured onto a synthetic hydrophilic dressing were applied with (N = 9) and without (N = 11) an acellular dermal matrix to full-thickness skin defects on athymic mice. Host cells progressively repopulated the acellular dermal component of the grafts. All animals with dermal matrix revealed fully differentiated epidermis by postoperative day 21. Human keratinocytes persisted in all animals grafted with dermal matrix, compared to only 63.6% of those animals without a dermal component. Planimetric analysis revealed significantly reduced wound contraction (p = 0.016) in animals receiving the dermal matrix. Histologic, immunohistochemical, and electron microscopic analyses also were performed. These studies suggest that an acellular dermal matrix can effectively direct regeneration of normal skin morphology. PMID- 9404990 TI - Cultured allogeneic keratinocyte sheets accelerate healing compared to Op-site treatment of donor sites in burns. AB - Donor site treatment is a crucial issue in the treatment of extensive burns. In this single-blind, randomized study treatment of donor sites with a polyurethane dressing, Op-Site (Smith & Nephew, York, U.K.) is compared to treatment with allogeneic cultured keratinocyte sheets. Results show a mean healing time of 6.7 days with use of cultured keratinocyte sheets compared to mean healing time of 13.6 days with Op-Site treatment. Also, improvement in the comfort of patients as the result of less exudate formation and pain attenuation was noted. PMID- 9404991 TI - Hyaluronic acid delays or impedes reepithelialization? PMID- 9404992 TI - Measures of line jaggedness and their use in foods textural evaluation. AB - The jaggedness of signatures, such as the force-displacement curves of brittle cereal-based foods, can be a manifestation of physical processes. Consequently, it should be regarded as an inherent property of the system rather than noise that should be discarded. Changes in the degree of jaggedness can provide useful information on the system's performance or on how it is affected by external conditions, such as exposure to moisture. Quantitative assessment of the degree of jaggedness can be done by methods that include statistical analysis, conversion of the record into a Fourier power spectrum, and determination of its apparent fractal dimension. Most can be performed with microcomputers and commercially available software. It is recommended that the jaggedness of any experimental curve or signature be determined by at least two methods or algorithms simultaneously for mutual verification. The magnitude of any single measure of jaggedness is determined by the signature's fluctuations amplitude and frequency. While the amplitude effect can be accounted for by normalization, the frequency cannot. Consequently, only curves having about the same number of points at the pertinent interval can be meaningfully compared. There is, however, a mathematical method, at this time still empirical, that allows meaningful comparison of the jaggedness of signatures obtained at different sampling rates. When several or numerous jagged signatures are recorded simultaneously, as in the case of brittle particulates compressed in bulk, the resulting combined signature is considerably smoothed. This is primarily the result of averaging caused by mutual cancellation of peaks in different directions and reducing the relative weight of any large peak. However, there are mathematical methods, some partly empirical, that enable restoration of the jaggedness of the original signatures. PMID- 9404993 TI - Migration and sorption phenomena in packaged foods. AB - Rapidly developing analytical capabilities and continuously evolving stringent regulations have made food/package interactions a subject of intense research. This article focuses on: (1) the migration of package components such as oligomers and monomers, processing aids, additives, and residual reactants in to packaged foods, and (2) sorption of food components such as flavors, lipids, and moisture into packages. Principles of diffusion and thermodynamics are utilized to describe the mathematics of migration and sorption. Mathematical models are developed from first principles, and their applicability is illustrated using numerical simulations and published data. Simulations indicate that available models are system (polymer-penetrant) specific. Furthermore, some models best describe the early stages of migration/sorption, whereas others should be used for the late stages of these phenomena. Migration- and/or sorption-related problems with respect to glass, metal, paper-based and polymeric packaging materials are discussed, and their importance is illustrated using published examples. The effects of migrating and absorbed components on food safety, quality, and the environment are presented for various foods and packaging materials. The impact of currently popular packaging techniques such as microwavable, ovenable, and retortable packaging on migration and sorption are discussed with examples. Analytical techniques for investigating migration and sorption phenomena in food packaging are critically reviewed, with special emphasis on the use and characteristics of food-simulating liquids (FSLs). Finally, domestic and international regulations concerning migration in packaged foods, and their impact on food packaging is briefly presented. PMID- 9404994 TI - Unraveling the chemical identity of meat pigments. AB - This review examines the chemistry of nitrite curing of meat and meat products as it relates to the development of cured meat color and provides a detailed account of how nitrite-free processed meats could be prepared using the preformed cooked cured-meat pigment (CCMP). Thus, a chemical description of meat color, both raw and cooked, and characterization of nitrosylheme pigments follows. Based on electron paramagnetic resonance (EPR), visible and infrared spectroscopic studies, evidence has been provided to support the hypothesis that the chemical structure of the preformed CCMP is identical to that of the pigment prepared in situ after thermal processing of nitrite-cured meat and is in fact a mononitrosylheme complex. An appendix, which describes the basic principles of EPR spectroscopy used in the context of this review, is attached. PMID- 9404995 TI - Ictal and interictal ECD-SPECT for focus localization in epilepsy. AB - Forty-one ECD (Technetium-99m-ethyl cysteinate dimer) SPECT investigations were undertaken in the course of a presurgical diagnostic work-up in 23 patients with pharmacoresistant focal epilepsy. In 21 patients, both an ictal and interictal SPECT were conducted. In the patients receiving ictal SPECT the tracer was injected between 7 and 30 s after the seizure onset. Of the interictal SPECTs 17 of 23 showed focal hypoperfusion which was consistent in 17 cases (74%) with the area of the electrophysiological focus (EF) and 6 patients had a normal interictal SPECT. Of the ictal SPECTs 18 of 21 (86%) showed regional hyperperfusion, 18 of them in the same location as the EF. Ictal SPECT showed a hypoperfusion similar to that in interictal SPECT in another 3 patients. In these cases seizure duration was short (28-54 s), so that the tracer reached the brain postictally. Our results show that ictal ECD-SPECT is an effective method for demonstrating an epileptogenic focus. Possible reasons for false-negative ictal SPECT results are discussed. PMID- 9404998 TI - Distorted colour discrimination in Parkinson's disease is related to severity of the disease. AB - The Farnsworth-Munsell 100-hue test (FMT) may be used for measurement of colour discrimination and error scores of the FMT provide an unspecific biological marker for the distinction between parkinsonian patients (PP) and healthy controls. The aim of this study was to examine the possible association between distorted colour discrimination and disease severity in untreated "de novo" PP Error scores of the FMT were significantly (P<0.0001) elevated in PP compared to age- and sex-matched controls and correlated to severity of the disease. We conclude that impaired colour discrimination is related to pathophysiology of Parkinson's disease. But it remains unclear whether these alterations of colour vision reflect striatal dopamine deficiency or changes of retinal dopaminergic pathways in PP. PMID- 9404996 TI - Eclamptic encephalopathy: imaging and pathogenetic considerations. AB - Eclampsia is a rare condition peculiar to pregnant and puerperal women, characterized by clinical pre-eclampsia (hypertension, proteinuria, edema) and generalized seizures. Three cases of eclamptic encephalopathy are reported: CT and MRI demonstrated transient abnormalities in the cortical and subcortical regions of the posterior areas of the brain - namely, parieto-occipital lobes - associated with occasional involvement of basal ganglia and/or brainstem. Pathogenesis is still unclear. Strict similarity with the pathological findings characterizing hypertensive encephalopathy suggests that a focal impairment in cerebral autoregulation may be the cause of vasodilation and fluid extravasation leading to hydrostatic edema; selective involvement of posterior areas could be explained by their lesser degree of adrenergic innervation supporting circulatory autoregulation mechanisms. PMID- 9404997 TI - Reduced adhesion of PBMNCs to endothelium in methylprednisolone-treated MS patients: preliminary results. AB - Methylprednisolone (MP) is a synthetic steroid commonly used in the treatment of multiple sclerosis (MS) relapses. It has a wide spectrum of activities on immune cells: it might also act by preventing mononuclear cell/endothelium adhesion. We studied adhesion phenomena between cultured human umbilical vein endothelial cells (HUVECs) and PBMNCs (CD45+, CD14+) from 6 MS patients treated in vivo with MP. We also studied fluctuations in CD11a and CD18 levels on lymphocytes and monocytes, as well as changes in serum sICAM-1 and sVCAM-1 concentrations. After MP treatment, PBMNCs adhesion to endothelium decreased at 3 h, while it went back to baseline levels at 24 h. A tendency to increase in both CD11a and CD18 on the surface of lymphocytes was detected, while an increase in serum sVCAM-1 was seen at 3 h. PMID- 9405000 TI - An association of human T-cell lymphotropic virus type I infection with vascular dementia. AB - Subjects ranging in age from 50 to 89 years old, either with or without dementia were studied by both ELISA for anti-human T-cell lymphotropic virus type I (HTLV I) gag 100-130 antibody and by cranial CT in order to clarify the relationship between HTLV-I infection and dementia. The frequency of anti-HTLV-I antibody was found to be significantly higher in the patients with dementia (24/130, 18.5%) than in those without dementia (11/139, 7.9%) (P=0.0169). Among the various types of dementia, HTLV-I seropositivity was found to be significantly associated with vascular dementia (11/48, 23%) (P=0.0087), but not with Alzheimer type dementia. In addition, HTLV-I seropositivity was also associated with Babinski sign, and the severity of cerebral infarction, ventricular dilatation and periventricular lucency on CT. The presence of HTLV-I therefore appears to be one of the risk factors for vascular dementia in HTLV-I endemic areas. PMID- 9404999 TI - Role of radiosurgery in the management of cavernous sinus meningiomas. AB - OBJECTIVE: To provide our early experience and philosophy in the utility of radiosurgery in the management of cavernous sinus meningiomas. METHODS: Twenty five consecutive cases with cavernous sinus meningiomas treated between 1990 and 1995 were reviewed. Three cases were treated with gamma-knife radiosurgery, 15 with preceding surgery and gamma knife, 7 with surgery. Mean follow-up following radiosurgery and surgery were 34.8 and 25.4 months, respectively. RESULTS: The 5 year actuarial tumor control rate following radiosurgery was 85.7% and tumor remission rate was 61.4%. Permanent neurological deterioration after radiosurgery was seen in 1 case (5.9%), whereas newly developed or worsened neurological deficits permanently persisted in 59.1% of patients after surgery. There was a clear correlation between surgical radicality and postoperative morbidity rate. CONCLUSIONS: Gamma-knife radiosurgery is a valuable addition to surgical removal in the treatment of cavernous sinus meningiomas. Combination of non-radical resection and subsequent radiosurgery is recommended to improve treatment associated morbidity. PMID- 9405002 TI - Early CCT signs of supratentorial brain infarction: clinico-radiological correlations. AB - Early signs of brain infarction can be detected by modern CCT technology even within the first 6 h after stroke. Little is known about the prognostic significance of early infarction signs in CCT. We prospectively evaluated clinical and CCT findings of 95 consecutive patients with an acute ischemia in the territory of the middle cerebral artery. All patients were admitted to our stroke unit within 6 h after stroke. In 55 patients CCT was performed within 3 h, and in 40 cases between 3 and 6 h. In all patients the clinical findings were assessed by the Scandinavian Stroke Scale (SSS). The disability due to stroke was evaluated after 4 weeks by use of the modified Rankin Scale. We could demonstrate the following early signs of cerebral infarction: focal hypodensity (23.2%), obscuration of basal ganglia (12.6%), focal brain swelling (22.1%), hyperdense middle cerebral artery sign (HMCA; 11.5%). In 3 patients early edema led to ventricular compression, in 1 patient to midline shift. The occurrence of early infarction signs did not depend on the etiology of ischemia but was significantly associated with a severe neurological deficit at admission and an unfavourable disability status 4 weeks after stroke. Focal brain swelling and HMCA were often followed by extensive infarction lesions on the follow-up CCT. In conclusion, early signs of hemispheric brain infarction visible on CCT scans performed within 6 h after stroke are correlated with severe stroke and an unfavourable functional outcome. However, a substantial part of our patients had a benign course of the disease in spite of early CCT pathology. Decisions on therapy in individual patients therefore should not depend on early CCT findings exclusively. PMID- 9405001 TI - Neuropsychological deficits in asymptomatic atrial fibrillation. AB - OBJECTIVE: To assess the preclinical effects on cognitive functions of nonrheumatic atrial fibrillation (NRAF) in patients with negative history for cerebrovascular disease. MATERIALS AND METHODS: The study included 37 consecutive patients with chronic (n=16, mean age 65.3+/-6.6 years) or paroxysmal (n=21, mean age 58.3+/-9.5 years) NRAF and an equal number of control subjects in sinus rhythm, who were matched for age, education and presence of hypertension. A comprehensive neuropsychological battery including tests of attention, memory, language and visuospatial skills was administered. RESULTS: Patients with chronic NRAF showed significantly poorer performances in tasks exploring attention and verbal memory functions, while the paroxysmal group was significantly impaired in a long-term memory task. The neuropsychological findings were confirmed excluding from both groups patients with CT evidence of cerebrovascular damage. A small subgroup of patients was also submitted to cerebral MRI. CONCLUSION: Neurologically asymptomatic NRAF is related to a subclinical but significant impairment in attention and memory. These deficits could be produced by minor ischemic lesions due to microembolization, or by diffuse hypoxic damage due to hypoperfusion. PMID- 9405003 TI - Basilar branch disease presenting with progressive pure motor stroke. AB - OBJECTIVES: Isolated infarcts of the pons cause well definable neurological syndromes with distinct pathomechanism, clinical course and prognosis. PATIENTS AND RESULTS: We report 8 cases suffering from a pure motor hemiparesis that was severely progressive within the 1st 3 days and unresponsive to aspirin. A relatively good recovery was observed in all patients, however, stroke recurrence occurred in 2 cases within 3 months and resulted in pseudobulbar paralysis and tetraparesis. MRI displayed unilateral (n=6) and bilateral (n=2) ventromedial pontine infarctions (VPI). Angiographic evaluation (n=4) or color Duplex examination (n=4) revealed atherosclerotic lesions but no basilar artery occlusion. CONCLUSION: Although VPI due to basilar branch disease may clinically mimic a classical lacunar syndrome, it is related to a particular pathogenetic mechanism different from microangiopathy or embolism. In contrast to the MRI feature of lacunes, VPI typically extend to the basal surface of the pons. The progressive pattern, ending up in a relative uniform clinical picture, is probably caused by propagating thrombosis. PMID- 9405004 TI - Transverse myelitis following mumps in an adult -- a case report with MRI correlation. AB - This is a case report of the 2nd oldest patient reported in the literature with transverse myelitis after mumps, and the 1st with magnetic resonance imaging (MRI) correlation. He is a 38-year-old Chinese man presenting with bilateral lower limb weakness and numbness, and urinary retention starting 3 weeks after an attack of mumps parotitis. Clinically, there was mild lower limb paresis, absent plantar responses and reduced pain sensation below the umbilicus. MRI revealed cord swelling and increased T2 signal from T7 to T11. Cerebrospinal fluid showed 23 cells/mm3 and 55 mg protein/dl. He received a 5-day course of intravenous methylprednisolone 0.5 g/d. The sensory and motor deficits improved over 2 weeks; urinary symptoms improved over the next year. Transverse myelitis following mumps is recognizable clinically and radiologically, and potentially responsive to methylprednisolone. PMID- 9405005 TI - Selective vertical saccadic palsy from unilateral medial thalamic infarction: clinical, neurophysiologic and MRI correlates. AB - BACKGROUND: Impairment of vertical gaze has been attributed to lesions involving the neural structures at the mesodiencephalic level. OBJECTIVE: Eye movements were studied in a patient with a unilateral paramedian thalamic infarction documented by MRI. CASE DESCRIPTION: A 63-year-old man presented 3 days after sudden onset vertical diplopia and hypersomnia. Eye movements were studied with electro-oculography and revealed impairment of vertical saccades with sparing of the vertical vestibulo-ocular reflex, vertical pursuit, Bell's phenomenon and vertical optokinetic nystagmus. MRI scan revealed a circular zone of altered signal intensity, suggesting infarction, in the paramedian ventral part of the right thalamus. CONCLUSIONS: This case demonstrates that a unilateral lesion mainly affecting the dorsomedial nucleus of the thalamus can result in selective impairment of vertical saccades and suggests that the corticofugal fibers mediating vertical saccades traverse in the medial thalamus en route to the rostral midbrain. PMID- 9405006 TI - Seasonal variation of the abdominal aortic aneurysm rupture in southwestern Greece. AB - OBJECTIVE: To examine the hypothesis of seasonal variation in the rupture of abdominal aortic aneurysm in our region. DESIGN: Retrospective open study. SETTING: University Hospital, Greece. PATIENTS: Forty-six patients with abdominal aortic aneurysm admitted between 1991-1995. INTERVENTION: The month of admission was registered. MAIN OUTCOME MEASURES: The seasonal variation in the abdominal aortic aneurysm rupture. RESULTS: The majority of ruptures (78%) occurred during the months October and April, a phenomenon proven to be periodical (p<0.05). CONCLUSIONS: The abdominal aortic aneurysm rupture clearly showed seasonal variation. PMID- 9405007 TI - Late results of surgery for abdominal aortic aneurysm. AB - We investigated the long-term survival and quality of life (QOL) of patients after abdominal aortic aneurysm (AAA) repair, in a retrospective study of 216 patients who underwent surgery at Nagoya University Hospital between 1980 and 1992. There were 189 elective operations and 27 emergency operations (22 ruptures and 5 impending ruptures). Complete follow-up information was available for 91% of the 200 patients who survived the surgery. Excluding operative deaths there was no significant difference in survival between patients who underwent elective or emergency surgery. The survival for both groups was close to that of age- and sex-matched cohorts. A significantly shorter survival time was found in patients over 75 than in those who were 74 or less (p<0.05). Preoperative heart disease or hypertension did not affect long-term survival rate. At the time of the last follow-up, 70% of the patients were quite healthy, and approximately 20% continued to work. We conclude that long-term survival after AAA repair is as good as that of the general population, and that survivors enjoy a good quality of life. However, there may have been preoperative selection for low risk patients in this study. PMID- 9405008 TI - Pancreaticoduodenal artery aneurysm. Case report with literature review. AB - Pancreaticoduodenal artery aneurysms are quite rare; only 5 cases have been reported in the Chinese literature. The variable clinical presentations and high incidence of rupture make it difficult to diagnose and treat. In this paper we report a case of inferior pancreaticoduodenal artery aneurysm with literature review. The case was found incidentally during a cholecystectomy and was later successfully treated by elective surgery. PMID- 9405009 TI - Monograft reconstruction of all the major aortic arch trunks. AB - Male aged 56 years, a smoker of 60 cigarettes daily, with multifocal disease, presented with vertigo, headaches, syncopal crises and intermittent claudication in both lower extremities. Angiography revealed pre-occlusive stenosis at the origin of the innominate artery, a significant degree of stenosis at the origin of the left common carotid and occlusion of the left subclavicular artery with steal syndrome as well as injury to the aortoiliac system. Revascularisation of all the branches of the aortic arch concomitantly was achieved with the technically simple monograft method using only one Dacron double velour 8 mm prosthesis, restoring circulation to both the cerebral and upper extremities without postoperative complications. Eight years later the subjective clinical findings have remained unchanged. Angiography revealed good function of the graft and subjectively the patient remains in satisfactory condition. PMID- 9405010 TI - Localised thrombus in the distal aortic arch: its aetiology analysed by three dimensional mould model of the thoracic aorta. AB - Isolated thrombus of the thoracic aorta without aneurysmal change or dissection is a relatively rare event. A case presenting with aortic thrombus and successive distal embolism is reported herein and the aetiology of the thrombus of the distal aortic arch is analysed by a three dimensional aortic model. A 61-year-old man suffered acute ischaemia in his right leg on January 26, 1994. Enhanced computed tomography (CT) showed a localized thrombus in the distal aortic arch expanding towards the descending aorta, and partial infarction of the left kidney and the spleen. Angiography demonstrated abrupt occlusion of the right superficial femoral artery. Immediate anticoagulation with heparin and coumadin was administered, and the thrombus in the aorta disappeared following 1 month of this medical treatment, leaving the renal and splenic infarction unchanged. The unresolved occlusion of the superficial femoral artery and the popliteal artery was treated with a bypass from the right superficial femoral artery to the peroneal artery using a reversed saphenous vein graft. The mould model of the thoracic aorta was reconstructed from CT, and the thrombus was found to be at the most distal and medial site of the lesser curvature of the aortic arch. This specific location is referred mostly as the site for thrombus formation in the literature. The case is reported briefly and the risk of this specific region of the thoracic aorta for thrombus formation is discussed using this mold model. PMID- 9405011 TI - Mycotic aneurysm of the bilateral tibioperoneal trunks associated with bacterial endocarditis: a case report. AB - Since antibiotics have been widely used in the treatment of bacterial endocarditis, mycotic aneurysms caused by septic emboli have become extremely rare. We report the case of a 34-year-old man who had mycotic aneurysms in the tibioperoneal trunks of both legs six weeks after he had a mitral valve replacement due to Streptococcus viridans endocarditis. Color Doppler sonography was used to diagnose pseudoaneurysms in both legs. Surgical treatment included the closures of the orifices of both aneurysms in both legs and arterial reconstructions were not required. In the literature, however, the location of mycotic aneurysms in peripheral arteries related to endocarditis were usually reported to be in the upper extremities or femoral arteries. Thus, we present the extremely rare case of mycotic aneurysms because the aneurysms occurred in the infrapopliteal vessel and developed in both legs. PMID- 9405012 TI - Grossly punched-out lesions in the aorto-iliac region can be histologically classified as false, pseudo-false, or disguised aneurysm. AB - Aneurysms are morphologically classified as true or false based on the nature of their walls. True aneurysms are composed of all or parts of layers of the vessel. False aneurysms are the result of rupture and their walls have only fibrous tissues. The orifice of false aneurysms is narrow relative to the aneurysmal diameter and thus they are grossly or angiographically referred to as punched-out lesions. Hence false aneurysms present with punched-out lesions, but in reverse, are all of punched-out lesions false aneurysms? We experienced some cases of punched-out lesions which histologically contained traces of elastin, and the purpose of this report was to histologically investigate grossly punched-out lesions. We examined 671 elderly autopsy cases, and a total of 21 grossly punched out lesions in the aorto-iliac region were selected. They were histologically classified as false, "pseudo-false", or "disguised" aneurysm. False aneurysms were found in 3 patients (0.45%), and were histologically mycotic. A total of 5 "pseudo-false" aneurysms were found in 3 patients (0.45%). They histologically contained traces of elastin, and thus they were categorised in true aneurysms. A total of 13 "disguised" aneurysms were found in 6 patients (0.89%). They were true fusiform aneurysms with an eccentric thrombus, on which a fibrin-cap formed a narrow orifice. Partial sections are insufficient for diagnosis; cross-sections are necessary. To the best of our knowledge, there have been no reports of "pseudo-false" or "disguised" aneurysms in the aorto-iliac region. PMID- 9405013 TI - Effect of lightweight compression stockings on venous haemodynamics. AB - OBJECTIVE: To investigate the effect of lightweight graduated elastic stockings on venous haemodynamics. DESIGN: The amount of reflux and function of the calf muscle pump were evaluated before and after the application of lightweight graduated compression stockings using air-plethysmography. Each patient acted as his own control and the Wilcoxon rank sum test was used. SETTING: Vascular laboratory of a teaching hospital. SUBJECTS: 19 female patients (20 limbs) with moderate varicose veins. MAIN OUTCOME MEASURES: The haemodynamic parameters: amount of reflux (VFI), ejection fraction (EF) of the calf muscle pump after one tiptoe exercise, residual volume fraction (RVF) after 10 tiptoes and venous volume (VV) were determined for each patient with and without the three strengths of stocking (7, 10, 14 mmHg at the ankle) using air-plethysmography. RESULTS: The mean VFI decreased from 5.7 ml/sec without stockings to 4.6+/-2.2, 3.9+/-2.3, and 3.4+/-1.8 with stockings of 7, 10 and 14 mmHg respectively (p<0.0002). Similarly the RVF showed a significant decrease with all three stockings from the initial value 42.3% to 36.3, 34.4 and 31.5 respectively (p<0.03). EF showed an increase from 49.2% to 51.4, 50.9 and 56, but only with the latter was the increase significant (p<0.02). VV decreased from 118.8 ml to 113.6+/-24.4 (p>0.05), 104.2+/-22.8 and 109.1+/-27.4 (p<0.008) with 10 and 14 mmHg. CONCLUSIONS: The results indicate that lightweight compression stockings can have a significant effect on venous haemodynamics. They decrease the residual volume fraction and by inference ambulatory venous pressure. This is the result of an increase in the ejection fraction with a decrease of reflux. The results offer a possible physiological explanation on the relief of symptoms experienced when patients with varicose veins wear lightweight stockings. PMID- 9405014 TI - The effect of mobilisation of patients during treatment of thromboembolic disorders with low-molecular-weight heparin. AB - OBJECTIVE: To elucidate the risk of pulmonary embolism (PE) in patients with deep vein thrombosis (DVT) who are kept walking with compression bandages. EXPERIMENTAL DESIGN: Perfusion/ventilation scanning of the lungs was performed at admission and after 10 days of treatment. SETTING: General community hospital. PATIENTS: A total of 631 consecutive patients were studied (upper limit of the thrombi: iliofemoral vein, n=212; femoral or popliteal vein, n=302; lower leg, n=117). The patients received different dose regimens of low-molecular-weight heparin (dalteparin) subcutaneously. RESULTS: The study revealed that the prevalence of PE at baseline was between 45.1% and 51% (95% CI 38.2-55.2 and 45.2 56.8% respectively) in patients with proximal DVT, and 31.9% (95% CI 23.6-41.2%) in those with DVT restricted to the lower leg. The majority of these cases of PE were completely asymptomatic. The incidence of a new PE, revealed by a second lung scan on day 10 after admission, was 7.0% (95% CI 3.9-11.4%) in patients with iliofemoral DVT, 5.5% (95% CI 3.2-8.7%) in those with femoropopliteal DVT and 2.7% (95% CI 0.6-7.6%) in those with lower-leg DVT. These incidence rates for new PEs were significantly lower than the rates previously reported (p<0.01). The fatality rate was also lower compared with the literature: one patient suffered a fatal PE (0.2%; 95% CI 0-0.9%), four patients died from malignant tumours, and one from pneumonia. The frequency of malignant tumours was greater in this study than in the literature (23% in patients with iliofemoral DVT, 14% in those with femoropopliteal DVT and 9% in those with DVT of the lower leg). CONCLUSION: Mobile patients with DVT do not need bed-rest. Low-molecular-weight heparin s.c., compression bandages and walking exercises make home-treatment of DVT feasible. PMID- 9405015 TI - Venous thromboembolism in the State of Minas Gerais and its projection to Brazil: study based in 2,331,353 hospitalisations. AB - MATERIALS AND METHODS: This study had as its objective an estimate of the prevalence of venous thromboembolism (VTE) in the State of Minas Gerais and its projection to Brazil, based on an analysis of 2,331,353 hospitalisations in the period of January 1994 to November 1995. RESULTS: Cardiovascular diseases, except for surgical cases, were responsible for 279,982 hospitalisations. The target population were the users of the Brazilian state health system. The evaluation, through clinical diagnosis, demonstrated the high incidence of VTE in patients over 50. The hospitalisation rate and mortality rate, were compared with those presented in studies in the United States and Europe. CONCLUSIONS: The importance of awareness of thrombosis and use of prophylactic methods to reduce the risks of serious complications of VTE. PMID- 9405016 TI - Development of vasa vasorum in the arterially implanted autovein bypass graft and its anastomosis in the dog. AB - Using stereomicroscopy, light microscopy, and scanning electron microscopy, we investigated the development of vasa vasorum in the proliferated neointima of the autovein graft and its anastomoses implanted in the canine femoral artery against a background of poor distal runoff. In the stereomicroscopic examination, a microfil silicone rubber compound (MF) was injected transluminally or via perivascular vasa, and the vascular specimen was prepared for clearing by immersion in a methyl-salicylate solution. Vessel interstices filled with MF were found adjacent to the suture materials within 5 days of grafting. Fourteen days after implantation, luminally originating vasa vasorum were often visible in the neointima along the suture line and distributed into the media and adventitia connecting to the original vasa vasorum. At 6 months or more after grafting, many orifices of luminally originating vasa vasorum were seen along the suture line of both proximal end-to-end and distal end-to-side anastomoses and distributed into the thickened neointima forming a vasa network when the neointima had proliferated to over 250 microm in depth. On the other hand, some clefts filled with MF were found in mural thrombi deposited on the vascular sinus of the graft within 5 days, and these appeared to be one of the sources of luminally originating vasa vasorum on the graft distant from the suture line. Moreover, the development of numerous vasa vasorum was constantly demonstrated in the neointima when it had proliferated to over 250 microm in depth. PMID- 9405018 TI - Biological activities of thyroid hormone receptors. PMID- 9405017 TI - Anplag, a selective 5-HT2 receptor antagonist, reduces stenosis induced by balloon injury in the hypercholesterolaemic rabbit. AB - The purpose of this study was to examine the effect of anplag, a 5-HT2 receptor antagonist, on intimal hyperplasia (IH) after balloon injury. In 39 male New Zealand white rabbits, the left carotid artery was denuded of endothelium by an inflated balloon. The injury not only damaged the endothelium, but also caused damage to underlying smooth muscle cells. Twelve rabbits fed normal chow were divided into 2 groups: Group A; anplag was not administered, and Group B; anplag, 100 mg/kg, was given by gavage, including on the day of ballooning. The remaining 27 rabbits were fed chow containing 1% cholesterol and divided into 3 groups: Group C; anplag was not administered, Group D; anplag, 100 mg/kg, was given orally except on the day of ballooning, and Group E; anplag, 100 mg/kg, was given orally including on the day of ballooning. IH in Group B was 13.2% less than that in Group A and IH in Group D was 11.7% less than that in Group C. However, neither of these differences was significant. IH in Group E was significantly less (by 40.1%) than that in Group C. In conclusion, anplag significantly reduced IH after balloon injury in the hypercholesterolaemic rabbit by inhibiting proliferation of smooth muscle cells and cholesterol-uptake by macrophages when it was administered immediately before the procedure. PMID- 9405019 TI - Expectation bias with respect to growth hormone therapy in Turner syndrome. PMID- 9405020 TI - Testicular activin--too hot to handle? PMID- 9405021 TI - Lipoprotein(a) and growth hormone: is the puzzle solved? PMID- 9405022 TI - Polygenic models of non-insulin-dependent diabetes mellitus. PMID- 9405023 TI - New data on nuclear hormone receptor cofactors suggest a control of transcriptional repression by hormone-dependent chromatin remodelling. PMID- 9405024 TI - The effect of recombinant human GH replacement therapy on lipoprotein(a) and other lipid parameters in adults with acquired GH deficiency: results of a double blind and placebo-controlled trial. AB - The effect of GH substitution on serum lipids and lipoproteins, and in particular lipoprotein(a) (Lp(a)), was investigated in 32 adults with postoperative (acquired) GH deficiency as part of a double-blind, placebo-controlled trial. Seventeen men and fifteen women, aged 18-59 years (mean 42 years) from two centres (Hannover and Gottingen) were randomly assigned to two groups. Group P (placebo) received placebo for the first 12 months and recombinant human GH (rhGH) for the following 12 months (open phase). Group V (verum) was treated with rhGH for two consecutive periods of 12 months each. The target dose of rhGH was 2 U/m2 per day, given subcutaneously daily at bedtime. Serum concentrations of Lp(a), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C) and triglycerides were determined at 0, 12 and 24 months. In group V, Lp(a) increased significantly in the first as well as in the second year of treatment (P=0.02, 12 months versus baseline; P=0.016, 24 months versus 12 months). In contrast, Lp(a) levels remained unchanged in group P during the first 12 months (P=0.826 versus baseline), but increased significantly (P=0.002) during the second year, when all patients were administered GH. The increase in Lp(a) after 12 months of rhGH replacement therapy in all 32 patients was significant for Lp(a) baseline concentrations both above and below 20 mg/dl and was independent of the apolipoprotein(a) isoforms. Total and LDL-C decreased significantly after 1 year and triglycerides after 2 years in group V. A significant reduction was also observed in the TC/HDL-C and the LDL-C/HDL-C ratios. Our study shows an unfavorable effect of rhGH replacement therapy on Lp(a) levels, which is, however, counteracted by a favorable effect of rhGH on TC, LDL-C and the TC/LDL-C and LDL-C/HDL-C ratios. PMID- 9405025 TI - Insulin-like growth factors and their binding proteins in pleural fluid. AB - We investigated the expression and potential regulatory role of insulin-like growth factors (IGFs) and their specific binding proteins (BPs) in tuberculous and nontuberculous pleuritis. By using a radioimmunoassay after acid gel filtration chromatography, we found that mean concentrations of IGF-I were 211.9 +/- 20.2 microg/l and 203.2 +/- 31.1 microg/l in pleural fluid of 14 patients with tuberculous pleuritis and 9 patients with malignant pleuritis respectively. These values were near those in serum of the same patients (221.3 +/- 19.5 microg/l and 204.6 +/- 21.0 microg/l respectively). By using a specific protein binding assay, we found that mean concentrations of IGF-II were 345.3 +/- 61.0 microg/l and 167.6 +/- 22.7 microg/l in tuberculous and malignant pleural effusions respectively. These values were significantly lower than those in serum of the same patients (628.3 +/- 79.0 microg/l, P<0.025 and 532.0 +/- 85.9 microg/l, P<0.025 respectively). Because bioavailability and bioactivity of IGFs may be regulated by their binding to IGFBPs, we studied IGFBP patterns in the pleural fluid of 6 patients with tuberculous pleuritis. As assessed by Western ligand blotting the levels of IGFBP-1 and IGFBP-2 were increased whereas those of IGFBP-3 were decreased in pleural fluid in comparison with serum. The decrease in IGFPB-3 levels reflected increased proteolysis, as assessed by Western immunoblotting. In spite of this presence of IGFBPs, IGFs could be responsible for the local biosynthesis of 1.25-dihydroxyvitamin D (1,25-(OH)2D) since pleural fluid levels of both IGF-I and IGF-II significantly correlated with those of 1,25 (OH)2D. These results indicate that IGFs are detectable in pleural fluid and may contribute to control the activity of 25-hydroxyvitamin D-1alpha hydroxylase in tuberculous pleuritis. PMID- 9405026 TI - Allelic variations in the human growth hormone-1 gene promoter of growth hormone deficient patients and normal controls. AB - OBJECTIVE: Isolated growth hormone deficiency (IGHD) type IB is suggested to be more probably due to alterations in the genes directly involved in the hypothalamo-pituitary axis and/or in the specific transcriptional regulation (cis trans coupling) of the hGH-1 gene than to alterations in the gene itself. In this study we analyzed the hGH-1 gene promoter region for structural alterations and allelic variations. METHODS: The hGH-1 gene promoter region was analyzed by PCR, cycle sequencing and direct-blotting electrophoresis in a total of 212 individuals including 113 patients with IGHD type IB, 21 unaffected family members and 78 normal controls. RESULTS: Twenty-two sequence variation sites were identified. Of these, 14% were located around the region of -1075bp, 77% between 550bp and the translational start site (+1bp) and 9% within the first intron. Only one variation site affected a characterized cis-acting element, namely that of NF-1. Importantly, all the variations found in patients were also observed in non-affected family members as well as in normal unrelated controls. CONCLUSIONS: These findings imply that it is not a single variation within the GH-1 gene promoter, and therefore in the cis-acting elements, which causes IGHD. However, we can not exclude the possibility that combinations of variations might perturb expression. Furthermore, these data illustrate the normal heterogeneity of the GH 1 gene promoter region, a fact that has to be borne in mind whenever transcriptional studies are performed. PMID- 9405027 TI - Immunodetection of G proteins in human pituitary adenomas: evidence for a low expression of proteins of the Gi subfamily. AB - G proteins mediate signal transduction in a variety of cell systems. As the expression of these proteins has not yet been investigated in detail in human pituitary tumors, we evaluated the presence of G proteins in a series of tumors including six non-functioning adenomas, five GH-secreting adenomas, three prolactinomas and one TSH-secreting adenoma, using immunoblotting and immunohistochemistry. By immunoblotting, Gi1/2alpha was undetectable in six and barely detectable in nine tumors. A similar pattern of expression was observed by probing with the antibody to Gi3alpha, which detected a very weak band in 11 tumors and no protein in four. In contrast, using large amounts of membrane proteins (40 microg), both Gi1/2alpha and Gi3alpha were detected, although at very low levels, in the negative tumors. The low expression of these proteins appeared to be specific to tumoral tissues, as both Gi1/2alpha and Gi3alpha were abundant in normal human and rat pituitary. In all tumors, Go alpha, the two Gs alpha forms, Gq/11 and G beta were present in significant amounts. Semiquantitative analysis indicated that Gs alpha was clearly detected when 2.5 microg loaded proteins were used, whereas Gi1/2alpha and Gi3alpha were barely detected with 5 microg. By immunofluorescence, all tumors studied were markedly positive for Gs alpha that was immunolocalized at the cell periphery, whereas they showed a weak positivity for Gi1/2alpha and Gi3alpha. The study is the first to provide evidence for a low expression of Gi proteins, which are involved in the transduction of inhibitory signals, in pituitary adenomas. PMID- 9405028 TI - Longitudinal study of fasting proinsulin in 148 siblings of patients with insulin dependent diabetes mellitus. Study Group on Childhood Diabetes in Finland. AB - OBJECTIVE: To follow proinsulin immunoreactive material (PIM) in healthy siblings from the time of diagnosis of insulin-dependent diabetes mellitus (IDDM) in the proband, for at least 2 years. DESIGN AND METHODS: The study comprised 148 siblings representing 112 families. The siblings were recruited from the nationwide 'Childhood Diabetes in Finland' study and tested for immunological markers. If a sibling was found positive for islet cell antibodies (ICA) or insulin autoantibodies (IAA), PIM sampling was extended beyond 2 years. RESULTS: Of the 148 siblings, 12 developed IDDM 3-53 months after the diagnosis in the proband. Eleven of these siblings exhibited initially normal PIM concentrations. In nine siblings, samples were available both more than 6 months and during the last 6 months before the diagnosis of IDDM; PIM concentrations increased in seven, remained unchanged in one, and decreased in one in the period up to the diagnosis of IDDM (P < 0.05). Median PIM concentration did not change significantly during the examination period of 2 years in the 136 siblings who did not contract IDDM. Constantly increased PIM concentrations were found in 12 of the 136 siblings who did not develop IDDM. These 12 siblings were all ICA negative. CONCLUSION: In healthy siblings of IDDM patients exhibiting an initially low PIM concentration, an abrupt increase in PIM seems to precede the clinical manifestation of IDDM within 0-6 months. However, there were too few patients available to close follow-up to allow calculation of any predictive value of this increase. Persistently increased PIM concentrations were present in some healthy siblings who did not develop IDDM. The reason for that finding remains unclear, but it could be associated with previous B cell damage. PMID- 9405029 TI - Long-term vitamin D3 supplementation may have adverse effects on serum lipids during postmenopausal hormone replacement therapy. AB - OBJECTIVE: The positive short-term effects of postmenopausal hormone replacement therapy (HRT) on serum lipids are well known, but it has been suggested that they vanish with time. Cholecalciferol (vitamin D3) is widely used to prevent postmenopausal osteoporosis but the influence of vitamin D3 on serum lipids is poorly known. The long-term effects of HRT and vitamin D3 on the concentrations of serum lipids were studied in a population-based prospective 3-year study. DESIGN AND METHODS: 464 women were randomized into four treatment groups: (i) HRT (sequential combination of 2 mg estradiol valerate and 1 mg cyproterone acetate), (ii) Vit D3 (vitamin D3 300 IU/day), (iii) HRT+Vit D3 (both as above), (iv) placebo (calcium lactate 500 mg/day). RESULTS: 320 women completed the study. After three years of treatment, serum concentrations of low density lipoprotein (LDL) cholesterol decreased in the HRT group (10.1%, P<0.001) and the HRT+Vit D3 group (5.9%, P=0.005), increased in the Vit D3 group (4.1%, P=0.035) but remained unchanged in the placebo group. The concentrations of total cholesterol decreased by 5.8% in the HRT group (P<0.001) and by 3.3% in the HRT+Vit D3 group (P=0.023), but did not change in the other two groups. Serum concentrations of high density lipoprotein (HDL) cholesterol decreased in the Vit D3 group (5.2%, P=0.001), HRT+Vit D3 group (3.7%, P=0.046), and the placebo group (4.5%, P=0.006) but did not change significantly in the HRT group. The HDL/LDL ratio increased in the HRT group (10.5%, P=0.006) and decreased in the Vit D3 group (10.5%, P<0.001) whereas no changes occurred in the other two groups. In addition, serum triglycerides increased similarly in all groups (14.0-18.8%, P<0.05-0.001). CONCLUSIONS: Our results confirm the positive long-term effect of HRT with sequential estradiol valerate and cyproterone acetate on serum lipid concentrations. In addition, the results suggest that vitamin D3 supplementation may have unfavorable effects on lipids in postmenopausal women. Pure vitamin D3 treatment was associated with increased serum LDL cholesterol. Furthermore, the beneficial effects of HRT on serum LDL cholesterol content were reduced when estradiol valerate was combined with vitamin D3. However, the relevance of these associations to cardiovascular morbidity remains to be established. PMID- 9405030 TI - Anti-pancreatic autoimmunity and Graves' disease: study of a cohort of 600 Caucasian patients. AB - The aim of this study was to investigate the frequencies of clinical diabetes and humoral markers of anti-pancreatic autoimmunity in a homogeneous population of 600 Caucasian patients with recently diagnosed Graves' disease (GD), in order to characterize the specific features of this group of endocrine patients among subjects at risk of diabetes. Ten were already diabetic at GD diagnosis. Among the 590 non-diabetic patients, 29 had islet cell antibodies (ICA), including 15 with low titre ICA and only 1 ICA-positive subject with a familial history of diabetes. Twenty-four patients had insulin autoantibodies, including three in association with ICA. Glutamic acid decarboxylase (GAD)/64 kDa antibodies were found in 16 of the 150 tested sera, including 13 of the 29 ICA-positive sera. Four ICA-positive patients displayed 37/40 kDa antibodies, including three in association with GAD/64 kDa antibodies. During follow-up, one of the ICA-positive patients developed insulin-dependent diabetes, 14 years after the GD diagnosis. To summarize, this anti-pancreatic autoimmunity study was focused on a large but specific and homogeneous group of subjects at risk for diabetes: recently diagnosed GD patients. This population was characterized by a high prevalence of GAD/64 kDa antibodies but also by a low frequency of evolution towards diabetes and the slowness of the process which could be due to the fact that only a minority of subjects possessed a sufficient combination of anti-pancreatic markers at the same time. PMID- 9405031 TI - Molecular screening of both the promoter and the protein coding regions in the human ob gene in Japanese obese subjects with non-insulin-dependent diabetes mellitus. AB - OBJECTIVE: Although the molecular mechanism of obesity has been poorly understood, recent studies indicate that leptin plays a critical role in regulating both food intake and body weight. Because obesity decreases the sensitivity to insulin, the human ob gene is presumed to be one of the candidate genes for non-insulin-dependent diabetes mellitus (NIDDM) associated with obesity. Although the protein coding region in the ob gene has been screened for mutations, the promoter region and the non-coding first exon have not yet been studied. We investigated the involvement of the human ob gene, especially mutations at the promoter region and the non-coding first exon, in the development of NIDDM associated with obesity. SUBJECTS: The study group comprised 60 Japanese obese subjects with NIDDM (body mass index (BMI) 43.6 > or = BMI > or = 26.4, 29.0+/-0.41 (mean+/-S.E.M.)) and 24 obese individuals with impaired glucose tolerance (IGT) (30 > or = BMI > or = 26.4, 27.1+/-0.22). METHODS: Mutations at both the promoter region and all three exons in the human ob gene were screened by the single-stranded conformational polymorphism analysis. When aberrantly migrated bands were recognized, the PCR-amplified DNA fragment was directly sequenced. RESULTS: In the protein coding region a silent mutation in the second exon was detected. The non-coding first exon and the about 100 bp 5' flanking region of the gene which contains a proximal CCAAT/enhancer-binding protein site were screened, but no mutations were found. CONCLUSION: These results suggest that no mutations in either the promoter region at the about 100 bp 5'-flanking region of the gene, or in any of the three exons, are involved in the development of NIDDM or IGT associated with obesity. PMID- 9405032 TI - Diabetes insipidus from sarcoidosis confined to the posterior pituitary. AB - A young white man with new-onset central diabetes insipidus was discovered to have a posterior pituitary mass on magnetic resonance imaging. No other radiological abnormalities were noted in the anterior pituitary, infundibulum or hypothalamus. No other endocrinopathies were present: laboratory investigations showed normal basal concentrations of anterior pituitary hormones, including prolactin. The patient was suspected to have sarcoidosis affecting the posterior pituitary, because of the discovery of pulmonary sarcoidosis during his diagnostic evaluation. His symptoms of polydipsia and polyuria responded promptly to intranasal administration of 1-desamino-8-D-arginine vasopressin (DDAVP). The patient demonstrated complete regression of the posterior pituitary mass after a course of corticosteroid therapy. However, his diabetes insipidus persisted and he continues to need DDAVP treatment, currently at 12 months of follow-up. The resolution of the neurohypophysial mass was compatible with the diagnosis of pituitary sarcoidosis and this precluded the need for a transsphenoidal biopsy or surgery. PMID- 9405033 TI - Follistatin-like immunoreactivity in the cytoplasm and nucleus of spermatogenic cells in the rat. AB - Immunohistochemistry using an antiserum raised against the synthetic follistatin peptide (residues 123-134) was used, in the present study, to detect the stage specific appearance of immunoreactive follistatin in the rat testis. Follistatin immunoreactivity was not found in Sertoli and Leydig cells, while it was clearly detected in spermatogenic cells. Follistatin-like immunoreactivity was detected in the cytoplasm and nucleus of late pachytene spermatocytes. Although the reaction in the cytoplasm disappeared after meiosis, it continued to be intense in the nucleus from pachytene spermatocytes to round spermatids. This finding indicated that follistatin or its closely related peptide produced in late pachytene spermatocytes migrates from the cytoplasm to the nucleus. We subjected rat testis homogenate to affinity chromatography on a sulfate-cellulofine and anti-follistatin Cys (123-134)-Affi-Gel Hz column followed by reverse-phase HPLC and analyzed the resulting fractions by Western blotting using follistatin antiserum. Three major bands at 57, 45 and 39 kDa or four bands at 52, 44, 39 and 34 kDa were detected in crude preparations from rat testis homogenate, under reducing or non-reducing SDS-PAGE respectively. The protein from rat testis, which was recognized by anti-follistatin (123-134) antiserum, exhibited a characteristic pattern for follistatin on SDS-PAGE, i.e. slower migration under reducing conditions than under non-reducing conditions, suggesting that it was follistatin or its closely related protein. Follistatin or its closely related protein may be a stage-specific modulator of spermatogenesis. Since follistatin like immunoreactivity was not found in oocytes in any stage of development from embryonic to adult rats, it may act in an event specific to spermatogenesis, such as nuclear condensation. PMID- 9405034 TI - The activity of 17alpha-hydroxylase/C17-C20 lyase in the ovaries of immature hypophysectomized rats treated with recombinant FSH combined with various doses of human chorionic gonadotropin. AB - The activity of 17alpha-hydroxylase/C17-C20 lyase (17alpha-hydroxylase) in the ovaries and steroid hormone levels in the plasma were studied in immature hypophysectomized rats (IH-rats) treated with human recombinant follicle stimulating hormone (rec-FSH) alone or combined with various doses of human chorionic gonadotropin (hCG). Eleven days after hypophysectomy, rats were given rec-FSH alone (total dose, 40 IU), or combined with various doses (0.1 to 10 IU in total) of hCG, twice daily for 4 days. Plasma levels of progesterone, testosterone and estradiol, and ovarian 17alpha-hydroxylase activity were measured 18 h after the last injection. Histology showed that the ovaries treated with rec-FSH alone had large antral follicles, the theca interna cells of which were small in size compared with those treated with rec-FSH combined with hCG. The activity of 17alpha-hydroxylase in the ovaries of IH-rats treated with rec FSH alone was lower than that in the control IH-rats. It was markedly increased by treatment with rec-FSH combined with 1 or 10 IU hCG. Immunohistochemistry revealed that 17alpha-hydroxylase was localized only in the oocyte in the ovaries of control IH-rats and those treated with rec-FSH alone. When the IH-rats were treated with rec-FSH plus hCG, the number of immunopositive theca interna cells and interstitial cells, and their immunointensity were increased in an hCG dose dependent manner. The plasma estradiol levels in the IH-rats treated with rec-FSH alone were low, but significantly higher than those in the control IH-rats, and estradiol levels were noticeably elevated in IH-rats treated with rec-FSH plus hCG. These results suggest that a synergism between hCG (LH) and FSH is essential for follicular development and steroidogenesis in the ovaries, implying paracrine effects among granulosa cells, theca cells and probably oocytes. The functional significance of 17alpha-hydroxylase in the oocytes is discussed in relation to estradiol production. PMID- 9405035 TI - Organ-specific effects of 3,5,3'-triiodothyroacetic acid in rats. AB - In order to compare the effect of 3,5,3'-triiodothyroacetic acid (TRIAC) with those of triiodothyronine (T3) and thyroxine (T4), severely hypothyroid rats (n=56) were infused over 13 days with 1, 2 or 4 nmol/100 g body weight (BW) per day of T3 or 2, 4 or 8 nmol/100 g BW per day of T4 or TRIAC. The 8 nmol/100 g BW per day of T4 or TRIAC induced the same increase in resting metabolic rate, yet 4 nmol/100 g BW per day of T3 was more potent (P < 0.05). For inhibiting serum TSH levels, 2 nmol/100 g BW per day of TRIAC were significantly less active than 2 nmol/100 g BW per day of T4 or 1 nmol/100 g BW per day of T3 (TRIAC, serum TSH 35.5 +/- 5.7; T3 2.58 +/- 0.91; T4 2.12 +/- 0.59 ng/ml). At higher doses serum TSH and beta-TSH mRNA were unmeasurable. Using serum T3 levels as covariate, the action of T3 and T4 was identical on cardiac monodeiodinase type 1 (5'D1) activity and hepatic malic enzyme (Me) mRNA levels and similar for hepatic 5'D1 activity. The effect of TRIAC was compared with T3 by using increasing doses of 1, 2 and 4 nmol/100 g BW per day of T3 and 2, 4 and 8 nmol/100 g BW per day of TRIAC. ANOVA indicated that there was no major difference between the effects of the hormones since with increasing doses the response of hepatic 5'D1 mRNA levels and enzyme activity and Me mRNA remained parallel. However, when studying the effect on cardiac 5'D1 activity there was not only a difference for type of treatment (T3 > TRIAC) but this difference became greater with each increment in dose. Interestingly there was also only a small effect of TRIAC on increase in heart weight compared with T3 and T4. Brain cortex monodeiodinase type 2 (5'D2) was mainly inhibited by T4 infusions. Monodeiodinase type 3 (5'D3) was stimulated by T4, less so by TRIAC and least by T3, expressing probably the local T3 and TRIAC concentrations. In conclusion, despite apparently similar effects of TRIAC and T3 and T4 on hepatic parameters of thyroid hormone action, TRIAC differs considerably in terms of its effects on cardiac 5'D1 activity and possibly on other fundamental effects of thyroid hormones on the heart since heart weight increased significantly less with TRIAC than with T3 or T4. PMID- 9405036 TI - Species differences between male rat and ram pituitary somatostatin receptors involved in the inhibition of growth hormone secretion. AB - The sheep is a valuable model in which to study GH neuroregulation as its pattern of GH secretion is very close to that in humans. Furthermore, important differences in somatostatin (SRIH) action between rats and sheep have been found previously. Our goal was to compare in male rat and ram pituitaries the binding characteristics of somatostatin receptors and the effect of SRIH and 17 analogues on GH release. Using radioautography, SRIH binding was seen to be evenly distributed over the anterior pituitary of both species. In the binding assay, binding sites were three times more concentrated in rats than in sheep. Important interspecies differences in the action of SRIH and its analogues were found: they inhibited GH at lower concentrations in rats than in sheep. Seven peptides displayed greater inhibitory ability in sheep than in rats while three were more potent in rats. Agonistic potencies to inhibit GH release in rats were correlated with somatostatin receptors subtype 2 (sst2) affinities. Our data confirm and extend the quantitative differences between rat and sheep in SRIH inhibitory action on GH secretion and confirm that ligand-binding properties of a given receptor subtype cannot be extrapolated across species. PMID- 9405037 TI - Recognition between disordered states: kinetics of the self-assembly of thioredoxin fragments. AB - The disordered N- (1-73) and C- (74-108) fragments of oxidized Escherichia colithioredoxin (Trx) reconstitute the native structure upon association [Tasayco, M. L., & Chao, K. (1995) Proteins: Struct., Funct., Genet. 22, 41-44]. Kinetic measurements of the formation of the complex (1-73/74-108) at 20 degrees C under apparent pseudo-first-order conditions using stopped-flow far-UV CD and fluorescence spectroscopies indicate association coupled to folding, an apparent rate constant of association [kon = (1330 +/- 54) M-1 s-1], and two apparent unimolecular rate constants [k1 = (0. 037 +/- 0.007) s-1 and k2 = (0.0020 +/- 0.0005) s-1]. The refolding kinetics of the GuHCl denatured Trx shows the same two slowest rate constants. An excess of N- over C-fragment decreases the kon, and the slowest phase disappears when a P76A variant is used. Stopped-flow fluorescence measurements at 20 degrees C indicate a GuHCl-dependent biphasic dissociation/unfolding process of the complex, where the slowest phase corresponds to 90% of the total. Their rate constants, extrapolated to zero denaturant, k-1 = (9 +/- 3) x 10(-5) s-1 and k-2 = (3.4 +/- 1.2) x 10(-5) s-1, show m# values of (4.0 +/- 0.4) kcal mol-1 M-1 and (3.5 +/- 0.1) kcal mol-1 M-1, respectively. Our results indicate that: (i) a compact intermediate with trans P76 and defined tertiary structure seems to participate in both the folding and unfolding processes; (ii) not all the N-fragment is competent to associate with the C-fragment; (iii) conversion to an association competent form occurs apparently on the time scale of P76 isomerization; and (iv) the P76A variation does not alter the association competency of the C-fragment, but it permits its association with "noncompetent" forms of the N-fragment. PMID- 9405038 TI - Electrostatic channeling of substrates between enzyme active sites: comparison of simulation and experiment. AB - Recent simulation work has indicated that channeling of charged substrates between the active sites of bifunctional enzymes or bienzyme complexes can be significantly enhanced by favorable interactions with the electrostatic field of the enzymes. The results of such simulations are expressed in terms of transfer efficiencies, which describe the probability that substrate leaving the active site of the first enzyme will reach the active site of the second enzyme before escaping out into bulk solution. The experimental indicators of channeling, on the other hand, are factors such as a decrease in the transient (lag) time for appearance of the final product of the coupled enzyme reaction or a decrease in the susceptibility of the overall reaction rate to the presence of competing enzymes or competitive inhibitors. The work reported here aims to establish a connection between the transfer efficiencies obtained from simulation, with the above-mentioned experimental observables. This is accomplished by extending previously reported analytical approaches to combine the simulated transfer efficiency with the Michaelis-Menten kinetic parameters Km and Vmax of the enzymes involved; expressions are derived to allow both transient times and steady state rates to be calculated. These results are applied to the two systems that have been studied both theoretically and experimentally. In the first case, that of the bifunctional enzyme dihydrofolate reductase-thymidylate synthase (DHFR-TS), the experimentally observed decrease in transient times is found to be consistent with a transfer efficiency of >/=80%. In the second case, that of a citrate synthase-malate dehydrogenase fusion protein, a transfer efficiency of 73% is consistent with the experimental transient time measurements. Separate and independent analysis of the effects of adding the competing enzyme aspartate aminotransferase gives a transfer efficiency of 69%, in excellent agreement with the transient time results. The transfer efficiencies thus obtained from experimental results are in both cases in good agreement with those obtained from simulations that include electrostatic interactions. One important discrepancy between simulation and experiment, is however, found in the reported effects of adding a competitive inhibitor in the DHFR-TS system: qualitatively different results are expected from the theoretical analysis. A possible reason for this apparent contradiction is discussed. PMID- 9405039 TI - Determinants for substrate phosphorylation by p21-activated protein kinase (gamma PAK). AB - gamma-PAK, originally designated PAK I and subsequently identified as a member of the p21-activated protein kinase family, has been shown to have cytostatic properties and to be involved in maintaining cells in a nondividing state [Rooney, R. D., et al., (1996) J. Biol. Chem. 271, 21498-21504]. The determinants for phosphorylation of substrates by gamma-PAK have been identified by examining the kinetics of phosphorylation of a series of synthetic peptides patterned after the sequence KKRKSGL, which is the site phosphorylated by gamma-PAK in the Rous sarcoma virus nucleocapsid protein NC in vivo and in vitro. With these peptides, the recognition sequence for gamma-PAK has been shown to contain two basic amino acids in the -2 and -3 positions, as represented by (K/R)RXS, in which the -2 position is an arginine, the -3 position is an arginine or a lysine, and X can be an acidic, basic, or neutral amino acid. A basic amino acid in the -1 or -4 position improves the rate of phosphorylation by increasing the Vmax and decreasing the Km. An acidic amino acid in the -1 position increases the rate (2.5-fold), as does an acidic residue in the -4 position, although to a lower extent (1.6-fold). Proline in the -1 or +1 position has a deleterious effect and inhibits phosphorylation by gamma-PAK. The substrate requirements of protein kinases that recognize basic amino acids on the N-terminal side of the phosphorylatable residue such as cAMP-dependent protein kinase (PKA) and Ca2+/phospholipid-dependent protein kinase (PKC) have been compared with gamma PAK using the same peptides. An acidic residue in the -1 position negatively affects PKA and PKC; thus, peptides containing the sequence KRES can be used to identify gamma-PAK. PMID- 9405040 TI - Atomic resolution (0.94 A) structure of Clostridium acidurici ferredoxin. Detailed geometry of [4Fe-4S] clusters in a protein. AB - The crystal structure of the 2[4Fe-4S] ferredoxin from Clostridium acidurici has been solved using X-ray diffraction data extending to atomic resolution, 0.94 A, recorded at 100 K. The model was refined with anisotropic representation of atomic displacement parameters for all non-hydrogen atoms and with hydrogens riding on their parent atoms. Stereochemical restraints were applied to the protein chain but not to the iron-sulfur clusters. The final R factor is 10.03 % for all data. Inversion of the final least-squares matrix allowed direct estimation of the errors of individual parameters. The estimated errors in positions for protein main chain atoms are below 0.02 A and about 0.003 A for the heavier [4Fe-4S] cluster atoms. Significant differences between the stereochemistry of the two clusters and distortion of both of them from ideal Td tetrahedral symmetry can be defined in detail at this level of accuracy. Regions of alternative conformations include not only protein side chains but also two regions of the main chain. One such region is the loop of residues 25-29, which was highly disordered in the room temperature structure. PMID- 9405041 TI - Solution structure of the cellulose-binding domain of the endoglucanase Z secreted by Erwinia chrysanthemi. AB - Two-dimensional proton nuclear magnetic resonance spectroscopy has been used to determine the three-dimensional structure of the 62 amino acid C-terminal cellulose-binding domain (CBD) of the endoglucanase Z (CBDEGZ), secreted by Erwinia chrysanthemi. An experimental data set comprising 958 interproton nOe derived restraints was used to calculate 23 structures. The calculated structures have an average root-mean-square deviation between Cys4 and Cys61 of 0.91 +/- 0.11 A for backbone atoms and 1.18 +/- 0.12 A for the heavy atoms. The CBDEGZ exhibits a skiboot shape based mainly on a triple antiparallel beta-sheet perpendicular to a less-ordered summital loop. Three aromatic rings (Trp18, Trp43, and Tyr44) are localized on one face of the protein and are exposed to the solvent in a conformation compatible with a cellulose-binding site. Based on its original folding, we have been able to relate the CBD sequence to those of several domains of unknown function occurring in several bacterial chitinases as well as other proteins. This study also provides a structural basis for analyzing the secretion-related information specific to the CBDEGZ. PMID- 9405043 TI - Picosecond to hour time scale dynamics of a "three finger" toxin: correlation with its toxic and antigenic properties. AB - Toxin alpha from Naja nigricollis (61 amino acids, four disulfide bridges) belongs to the "three finger" fold family, which contains snake toxins with various biological activities and nontoxic proteins from different origins. In this paper, we report an extensive 1H and 15N NMR study of the dynamics of toxin alpha in solution. 15N relaxation, 1H off-resonance ROESY, and H-D exchange experiments allowed us to probe picosecond to hour motions in the protein. Analysis of these NMR measurements demonstrates that toxin alpha exhibits various time scale motions, i.e., particularly large amplitude picosecond to nanosecond motions at the tips of the loops, observable microsecond to millisecond motions around two disulfide bridges, second time scale motions around the C-N bonds of asparagine and glutamine side chains which are more or less rapid depending on their amino acid solvent accessibility, and minute to hour motions in the beta sheet structure. The less well-defined regions of toxin alpha solution structures are subject to important picosecond to nanosecond motions. The toxic site is organized around residues belonging to the rigid core of the molecule but also comprises residues exhibiting dynamics on various time scales. The Malpha1 epitope is subject to large picosecond to millisecond motions, which are probably modified by the interaction with the antibody. This phenomenon could be linked to the neutralizing properties of the antibody. PMID- 9405042 TI - Structural studies of the Escherichia coli signal transducing protein IIAGlc: implications for target recognition. AB - In Escherichia coli, the glucose-specific phosphocarrier protein of the phosphotransferase system (PTS), IIAGlc (IIIGlc in older literature), is also the central regulatory protein of the PTS. Depending upon its state of phosphorylation, IIAGlc binds to a number of different proteins that display no apparent sequence homology. Previous structural studies suggested that nonspecific hydrophobic interactions, specific salt bridges, and an intermolecular Zn(II) binding site contribute to the wide latitude in IIAGlc binding sites. Two new crystal forms of IIAGlc have been solved at high resolution, and the models were compared to those previously studied. The major intermolecular contacts in the crystals differ in detail, but all involve the hydrophobic active site of IIAGlc interacting with a hydrophobic patch on a neighbor and all are shown to be surprisingly similar to the physiologically relevant regulatory interaction of IIAGlc with glycerol kinase. In two crystal forms, a helix on one molecule interacts with the face of another, while in the other crystal form, the primary crystal contact consists of a strand of beta sheet that contributes to an intermolecular Zn(II) binding site with tetrahedral ligation identical to that of the zinc peptidase thermolysin. Thus, relatively nonspecific hydrophobic interactions combined with specific salt bridges and an intermolecular cation binding site (cation-promoted association) permit a regulatory protein to bind to target proteins that have little or no sequence or structural homology with one another. It is suggested that signal transduction by IIAGlc is a binary switch in which phosphorylation at the active site directly controls binding to target molecules. PMID- 9405044 TI - Determinants of substrate specificity in the superfamily of amino acid dehydrogenases. AB - The subunit of the enzyme glutamate dehydrogenase comprises two domains separated by a cleft harboring the active site. One domain is responsible for dinucleotide binding and the other carries the majority of residues which bind the substrate. During the catalytic cycle a large movement between the two domains occurs, closing the cleft and bringing the C4 of the nicotinamide ring and the Calpha of the substrate into the correct positioning for hydride transfer. In the active site, two residues, K89 and S380, make interactions with the gamma-carboxyl group of the glutamate substrate. In leucine dehydrogenase, an enzyme belonging to the same superfamily, the equivalent residues are L40 and V294, which create a more hydrophobic specificity pocket and provide an explanation for their differential substrate specificity. In an attempt to change the substrate specificity of glutamate dehydrogenase toward that of leucine dehydrogenase, a double mutant, K89L,S380V, of glutamate dehydrogenase has been constructed. Far from having a high specificity for leucine, this mutant appears to be devoid of any catalytic activity over a wide range of substrates tested. Determination of the three dimensional structure of the mutant enzyme has shown that the loss of function is related to a disordering of residues linking the enzyme's two domains, probably arising from a steric clash between the valine side chain, introduced at position 380 in the mutant, and a conserved threonine residue, T193. In leucine dehydrogenase the steric clash between the equivalent valine and threonine side chains (V294, T134) does not occur owing to shifts of the main chain to which these side chains are attached. Thus, the differential substrate specificity seen in the amino acid dehydrogenase superfamily arises from both the introduction of simple point mutations and the fine tuning of the active site pocket defined by small but significant main chain rearrangements. PMID- 9405045 TI - Crystal structures of the copper-containing amine oxidase from Arthrobacter globiformis in the holo and apo forms: implications for the biogenesis of topaquinone. AB - The crystal structures of the copper enzyme phenylethylamine oxidase from the Gram-positive bacterium Arthrobacter globiformis (AGAO) have been determined and refined for three forms of the enzyme: the holoenzyme in its active form (at 2.2 A resolution), the holoenzyme in an inactive form (at 2.8 A resolution), and the apoenzyme (at 2.2 A resolution). The holoenzyme has a topaquinone (TPQ) cofactor formed from the apoenzyme by the post-translational modification of a tyrosine residue in the presence of Cu2+. Significant differences between the three forms of AGAO are limited to the active site. The polypeptide fold is closely similar to those of the amine oxidases from Escherichia coli [Parsons, M. R., et al. (1995) Structure 3, 1171-1184] and pea seedlings [Kumar, V., et al. (1996) Structure 4, 943-955]. In the active form of holo-AGAO, the active-site Cu atom is coordinated by three His residues and two water molecules in an approximately square-pyramidal arrangement. In the inactive form, the Cu atom is coordinated by the same three His residues and by the phenolic oxygen of the TPQ, the geometry being quasi-trigonal-pyramidal. There is evidence of disorder in the crystals of both forms of holo-AGAO. As a result, only the position of the aromatic group of the TPQ cofactor, but not its orientation about the Cbeta-Cgamma bond, is determined unequivocally. In apo-AGAO, electron density consistent with an unmodified Tyr occurs at a position close to that of the TPQ in the inactive holo AGAO. This observation has implications for the biogenesis of TPQ. Two features which have not been described previously in amine oxidase structures are a channel from the molecular surface to the active site and a solvent-filled cavity at the major interface between the two subunits of the dimer. PMID- 9405047 TI - Energetics of heme binding to native and denatured states of cytochrome b562. AB - Cytochrome b562 is a four-helix bundle protein containing a noncovalently bound b type heme prosthetic group. For the first time, energetics of heme binding to an apocytochrome were measured by isothermal titration calorimetry. The heme is tightly bound to native apocytochrome b562, with a dissociation constant (Kd) of approximately 9 nM (DeltaG degrees = 11 kcal mol-1) at 25 degrees C. Unexpectedly, the thermally denatured apoprotein is also capable of specifically binding heme with modest affinity (Kd = 3 microM, DeltaG degrees = 7.6 kcal mol 1). This interaction results in the dependence of holocytochrome b562 stability on protein concentration in the submicromolar range. PMID- 9405046 TI - The alrestatin double-decker: binding of two inhibitor molecules to human aldose reductase reveals a new specificity determinant. AB - It is generally expected that only one inhibitor molecule will bind to an enzyme active site. In fact, specific drug design theories depend upon this assumption. Here, we report the binding of two molecules of an inhibitor to the same active site which we observed in the 1.8 A resolution structure of the drug Alrestatin bound to a mutant of human aldose reductase. The two molecules of Alrestatin bind to the active site in a stacked arrangement (a double-decker). This stack positions the carboxylic acid of one drug molecule near the NADP+ cofactor at a previously determined anion binding site and the carboxylic acid of the second drug molecule near the carboxy-terminal tail of the enzyme. We propose that interactions of inhibitors with the carboxy-terminal loop of aldose reductase are critical for the development of inhibitors that are able to discriminate between aldose reductase and other members of the aldo-keto reductase superfamily. This finding suggests a new direction for the introduction of specificity to aldose reductase-targeted drugs. PMID- 9405048 TI - The GTP effector site of ornithine decarboxylase from Lactobacillus 30a: kinetic and structural characterization. AB - A nucleotide effector site of the biodegradative form of ornithine decarboxylase from Lactobacillus 30a (OrnDC L30a) has been identified. OrnDC L30a activity at pH 8.0, where the enzyme is normally inactive, is stimulated by GTP and dGTP and to a lesser extent by GDP but not by ATP, CTP, or UTP. The pH profile indicates that activation by GTP is reflected by an increase in kcat/KM,orn (above pH 6.8), while Vmax remains constant over the pH range 4.0-9. 0. Scatchard plot analysis shows that GTP binds to OrnDC L30a at both pH 5.8 (KD = 0.11 microM) and pH 8.0 (KD = 1.6 microM), but unexpectedly, half-site binding is observed at the higher pH. The OrnDC L30a dodecamer dissociates into dimers at high pH in the presence or absence of GTP. The GTP binding site was located in difference electron density maps using low-resolution X-ray data. This represents a new type of GTP binding site. A model explaining the activation of OrnDC L30a by GTP is presented. PMID- 9405049 TI - X-ray structure of motor and neck domains from rat brain kinesin. AB - We have determined the X-ray structure of rat kinesin head and neck domains. The folding of the core motor domain resembles that of human kinesin reported recently [Kull, F. J., et al. (1996) Nature 380, 550-554]. Novel features of the structure include the N-terminal region, folded as a beta-strand, and the C terminal transition from the motor to the rod domain, folded as two beta-strands plus an alpha-helix. This helix is the beginning of kinesin's neck responsible for dimerization of the motor complex and for force transduction. Although the folding of the motor domain core is similar to that of a domain of myosin (an actin-dependent motor), the position and angle of kinesin's neck are very different from those of myosin's stalk, suggesting that the two motors have different mechanisms of force transduction. The N- and C-terminal ends of the core motor, thought to be responsible for the directionality of the motors [Case, R. B., et al. (1997) Cell 90, 959-966], take the form of beta-strands attached to the central beta-sheet of the structure. PMID- 9405050 TI - Dissection of the pH dependence of inhibitor binding energetics for an aspartic protease: direct measurement of the protonation states of the catalytic aspartic acid residues. AB - The catalytic activity and inhibitor binding energetics of enzymes are often pH dependent properties. Aspartic proteases comprise an important class of enzyme targets for structure-based drug design. We have performed a complete thermodynamic study of pepstatin binding to plasmepsin II, an aspartic proteinase found in Plasmodium falciparum, using isothermal titration calorimetry and circular dichroism. Thermodynamic parameters (DeltaG, DeltaH, DeltaCp, and DeltaS) were measured as functions of both pH and temperature. In the pH range from 4.5 to 7.0, pepstatin binding is accompanied by proton transfer between the solvent and the complex. We used thermodynamic proton linkage theory to derive both the pH-independent binding energetics for pepstatin and the number and pKa values of ionizable residues whose pKa values change during ligand binding. These residues were identified as the two catalytic aspartates, with pKas of 6.5 and 3.0, and His 164, with a pKa of 7.5, based on the three-dimensional structure of the pepstatin-plasmepsin II complex. At pH 5.0, where the protease has optimum activity, the proton transfer process contributes almost 40% of the total binding free energy change and the total charge of the active-site aspartic acid residues is -1. These experimental results provide direct measurement for the protonation states of the catalytic aspartates in the presence of bound ligands. Comparison of the thermodynamic and structural data for pepstatin binding with human cathepsin D, a lysosomal aspartic protease that shares 35% sequence identity with plasmepsin II, suggests that the energetic differences between these two proteins are due to a higher interdomain flexibility in plasmepsin II. PMID- 9405051 TI - Folding of an mRNA pseudoknot required for stop codon readthrough: effects of mono- and divalent ions on stability. AB - Unfolding of an mRNA pseudoknot that induces ribosome suppression of the gag gene stop codon in Moloney murine leukemia virus has been studied by UV hyperchromicity and calorimetry. The pseudoknot melts in two steps, corresponding to its two helical stems. The total enthalpy of denaturation is approximately 170 kcal/mol, approximately the value expected for the secondary structure. At low salt concentrations (<50 mM KCl) the unfolding transitions are not two-state, but they approach two-state behavior at higher salt concentrations. The structure is preferentially stabilized by smaller alkali metal ions (Li+ > Na+ > K+ > Rb+ > Cs+) and by NH4+; the same preferences are exhibited by one of the stems in the context of a hairpin. Divalent metal ions are not required to fold the pseudoknot but do stabilize it further. To examine divalent ion effects over a wide concentration range, urea was used to lower the RNA unfolding temperature and was shown not to affect characteristics of the pseudoknot unfolding in other respects. The pseudoknot binds divalent ions somewhat more tightly than a hairpin but shows only weak selectivity for different size ions. It is suggested that a region of "intermediate" divalent ion binding affinity, in between highly ligated specific sites and purely delocalized ion binding in character, is created by the pseudoknot fold but that nonspecific, delocalized ion binding contributes at least half the free energy of pseudoknot stabilization by Mg2+. PMID- 9405052 TI - Comparison of the physiologically equivalent proteins cytochrome c6 and plastocyanin on the basis of their electrostatic potentials. Tryptophan 63 in cytochrome c6 may be isofunctional with tyrosine 83 in plastocyanin. AB - The blue copper protein plastocyanin and the heme protein cytochrome c6 differ in composition and in structure but perform the same function in the photosynthetic electron-transport chain. We compare these two proteins on the basis of their electrostatic potentials in order to understand the structural basis of their functional equivalence. In the first approach, we use a monopole-dipole approximation of the electrostatic potentials to superimpose the proteins. The resulting alignment suggests that Tyr51 in cytochrome c6 corresponds to Tyr83 in plastocyanin. But since Tyr51 is not conserved in all known cytochrome c6 sequences, a physiological role of this residue is questionable. In a more sophisticated approach, we applied the recently-developed Fame (flexible alignment of molecule ensembles) algorithm, in which molecules are superimposed by optimizing the similarity of their electrostatic potentials with respect to the relative orientation of the molecules. On the basis of the Fame alignments of plastocyanin and cytochrome c6, we analyze the docking and the electron-transfer reactions of these two proteins with its physiological reaction partner cytochrome f. We derive functional analogies for individual amino acids in possible electron-transfer paths in the interprotein redox reactions. We identify two surface patches in cytochrome c6 that may be involved in electron-transfer paths. The hydrophobic patch with the exposed heme edge in cytochrome c6 may be equivalent to the hydrophobic patch with His87 in plastocyanin, whereas Trp63 in cytochrome c6 may be equivalent to Tyr83 in plastocyanin. An aromatic amino acid is present at the position of Trp63 in all known cytochrome c6 sequences. The electronic coupling between the heme and the copper site on the one side and several potentially important amino acid residues on the other is analyzed by the Pathways method. We have proposed recently that Lys65 of cytochrome f and Tyr83 of plastocyanin form a cation-pi system, which may be involved in a two-step mechanism of the electron-transfer reaction between these two proteins from higher plants. Now we corroborate this proposal by analyzing available amino acid sequences. PMID- 9405053 TI - Active site of dihydroorotate dehydrogenase A from Lactococcus lactis investigated by chemical modification and mutagenesis. AB - The flavin-containing enzyme dihydroorotate dehydrogenase (DHOD) catalyzes the oxidation of dihydroorotate (DHO) to orotate, the first aromatic intermediate in pyrimidine biosynthesis. The first structure of a DHOD, the A form of the enzyme from Lactococcus lactis, has recently become known, and some conserved residues were suggested to have a role in the active site [Rowland et al. (1997) Structure 2, 239-252]. In particular, Cys 130 was hypothesized to work as a base, which activates dihydroorotate (DHO) for hydride transfer. By chemical modification and site-directed mutagenesis we have obtained results consistent with this proposal. Cys 130 was susceptible to alkylating reagents, and mutants of Cys 130 (C130A and C130S) showed hardly detectable enzyme activity at pH 8.0, while at pH 10 the C130S mutant enzyme had approximately 1% of wild-type activity. Mutants of Lys 43, Asn 132, and Lys 164 were also constructed. Exchange of Lys 43 to Ala or Glu (K43A and K43E) and of Asn 132 to Ala (N132A) affected both catalysis and substrate binding. Expressed as kcat/KM for DHO, the deterioration of these three mutant enzymes was 10(3)-10(4)-fold. Flavin spectra of the mutant enzymes were not, like the wild-type enzyme, bleached by DHO in stopped-flow experiments, showing that they were deficient with respect to the first half-reaction, namely reduction of FMN by DHO, which was not rate limiting for the wild-type enzyme. The binding interaction between flavin and the reaction product, orotate, could be monitored by a red shift of the flavin absorbance in the wild-type enzyme. The C130A, C130S, and N132A mutant enzymes displayed similar capacity to bind orotate. In contrast, orotate did not change the absorption spectra of the K43 mutant enzymes, although it did inhibit their activity. All of the mutant enzymes, except K164A, contained normal levels of flavin. The results are discussed in relation to the structures of DHODA and other flavoenzymes. The possible acid-base chemistry of Cys 130 is compared to previous work on mammalian dihydropyrimidine dehydrogenases, flavoenzymes, which catalyze the reversed reaction, namely the reduction of pyrimidine bases. PMID- 9405054 TI - A novel assay for evaluating glycogenolysis in rat adipocytes and the inability of insulin to antagonize glycogenolysis in this cell type. AB - We report here on a novel procedure for measuring glycogenolysis in rat adipocytes. In this procedure, cells are incubated for 30 min at 37 degrees C with insulin or vanadate, and with [U-14C]glucose to label the glycogen pool with radioactive glucose. The cells are washed and preincubated for an additional 1 h, before being assayed. The extent of glycogenolysis is determined by the decrease in radioactivity in precipitated glycogen, which was quite substantial under experimental conditions facilitating glycogenolysis. From the assay, we determined the following. (a) Glycogenolysis is activated in rat adipocytes in response to lipolytic hormones (i.e. catecholamines and adrenocorticotropic hormone). (b) Other agents and conditions elevating intracellular adenosine 3',5' monophosphate levels (i.e. cholera toxin, dibutyryladenosine 3',5'-monophosphate, and isobutylmethylxanthine) also activate glycogenolysis. (c) Glycogenolysis (as opposed to lipolysis) is activated at concentrations of adrenocorticotropic hormone or isoproterenol 7-11-fold lower and at adenosine 3',5'-monophosphate concentrations 7-fold lower. (d) Calyculin A, a specific inhibitor of protein phosphatase 1, activates glycogenolysis as well. Calyculin A also activates lipolysis at an equimolar potency. (e) Insulin does not antagonize glycogenolysis in rat adipocytes. In conclusion, the assay allowed us to compare glycogenolysis to lipolysis within the same cell, and to find that the sensitivity to hormones and adenosine 3',5'-monophosphate was about 1 order of magnitude higher for glycogenolysis than for lipolysis. A more striking finding was the inability of insulin to antagonize glycogenolysis in the rat adipose cell, an effect which occurs readily in liver and muscle cells via protein phosphatase 1-activating machinery. This rules out a role for adipose protein phosphatase 1 activation in the mechanism by which insulin antagonizes lipolysis and supports the contention that the insulin effect in lowering adenosine 3',5'-monophosphate levels is the central mechanism by which insulin antagonizes lipolysis. PMID- 9405055 TI - Time-resolved protein phosphorescence in the stopped-flow: denaturation of horse liver alcohol dehydrogenase by urea and guanidine hydrochloride. AB - This study reports the implementation of room temperature protein phosphorescence in the stopped-flow technique. Time-resolved Trp phosphorescence can now be detected following rapid mixing of protein solutions with a time resolution of 10 ms and a sensitivity in terms of chromophore concentration down to 0.1 microM. Calibration tests with monomeric and multimeric proteins proved that in all cases the delayed emission is not affected by artefacts that could arise from either enrichment of trace impurities along the flow lines or deformation of the macromolecules by the shear stress of laminar flow. To illustrate the potential of Trp phosphorescence in the stopped-flow to detect the time evolution of protein conformation the interaction of urea and guanidine hydrochloride (GdnHCl) with the native structure of horse liver alcohol dehydrogenase (LADH) has been re examined under conditions of rapid denaturation. Remarkable differences in the action of the two denaturing agents has been confirmed by the phosphorescence lifetime (tauP) of the internal Trp residue (W314). Whereas in urea, up to 8 M, tauP is not minimally perturbed, in GdnHCl it decreases sharply and progressively from 800 ms down to 23 ms in 6 M solutions. Such reduction of tauP implies that in the region of W314 the polypeptide structure has become highly loose and flexible prior to the major unfolding transition. Therefore, denaturation of LADH in GdnHCl, as opposed to urea, proceeds from a partly unfolded intermediate conformation of the protein. Other characteristics of this intermediate state are a partial loss of tertiary structure, as revealed by the circular dichroism of the aromatics, and an almost complete inhibition of the catalytic activity. Control experiments with equimolar NaCl demonstrate that tauP, the tertiary structure and the catalytic activity are affected to a much smaller extent and that, therefore, salt effects do not account for the difference between urea and GdnHCl. Finally, measurements of the unfolding reaction emphazise that the kinetics of LADH denaturation are heterogeneous with both denaturing agents. From the constancy of tauP during the course of the reaction it is concluded that the multiphasic behavior is a manifestation of multiple unfolding pathways owing to a plurality of stable LADH conformations. PMID- 9405056 TI - Redox potentials and quinone reductase activity of L-aspartate oxidase from Escherichia coli. AB - l-Aspartate oxidase (EC 1.4.3.16) is a flavoprotein that catalyzes the first step in the de novobiosynthetic pathway to pyridine nucleotides both under aerobic and under anaerobic conditions. Despite the physiological importance of this biosynthesis particularly in facultative aerobic organisms, such as Escherichia coli, little is known about the electron acceptor of reduced L-aspartate oxidase in the absence of oxygen. In this report, evidence is presented which suggests that in vitro quinones can play such a role. L-Aspartate oxidase binds menadione and 2, 3-dimethoxy-5-methyl-p-benzoquinone with Kd values of 11.5 and 2.4 microM, respectively. A new L-aspartate:quinone oxidoreductase activity is described in the presence and in the absence of phospholipids, and its possible physiological relevance is discussed. Moreover, considering the striking sequence similarity between L-aspartate oxidase and the highly conserved family of succinate-fumarate oxidoreductases, the redox properties of L-aspartate oxidase were investigated in detail. A value of -216 mV was calculated for the midpoint potential of the couple FAD/FADH2 bound to the enzyme. This result perfectly explains why L aspartate oxidase may be considered as a very particular fumarate reductase unable to use succinate as the electron donor. PMID- 9405057 TI - Formation of a long-lived P+BA- state in plant pheophytin-exchanged reaction centers of Rhodobacter sphaeroides R26 at low temperature. AB - Femtosecond transient absorption spectroscopy in the range of 500-1040 nm was used to study electron transfer at 5 K in reaction centers of Rhodobacter sphaeroides R26 in which the bacteriopheophytins (BPhe) were replaced by plant pheophytin a (Phe). Primary charge separation took place with a time constant of 1.6 ps, similar to that found in native RCs. Spectral changes around 1020 nm indicated the formation of reduced bacteriochlorophyll (BChl) with the same time constant, and its subsequent decay in 620 ps. This observation identifies the accessory BChl as the primary electron acceptor. No evidence was found for electron transfer to Phe, indicating that electron transfer from BA- occurs directly to the quinone (QA) through superexchange. The results are explained by a model in which the free energy level of P+Phe- lies above that of P+BA-, which itself is below P*. Assuming that the pigment exchange does not affect the energy levels of P* and P+BA-, our results strongly support a two-step model for primary electron transfer in the native bacterial RC, with no, or very little, admixture of superexchange. PMID- 9405058 TI - Structural and functional investigations on the role of zinc in bifunctional rat peptidylglycine alpha-amidating enzyme. AB - Bifunctional peptidylglycine alpha-amidating enzyme (alpha-AE) catalyzes the two step conversion of C-terminal glycine-extended peptides to C-terminal alpha amidated peptides and glyoxylate. The first step is the ascorbate-, O2-, and copper-dependent hydroxylation of the alpha-carbon of the glycyl residue, producing an alpha-hydroxyglycine-extended peptide. The second step is the ascorbate-, O2-, and copper-independent dealkylation of the carbinolamide intermediate. We show that alpha-AE requires 1.1 +/- 0. 2 mol of zinc/mol of enzyme for maximal (S)-N-dansyl-Tyr-Val-alpha-hydroxyglycine dealkylation activity. Treatment of the enzyme with EDTA abolishes both the peptide hydroxylation and the carbinolamide dealkylation activities. Addition of Zn(II), Co(II), Cd(II), and Mn(II) partially restores carbinolamide dealkylation activity to the EDTA-treated enzyme. Addition of Co(II) produces the greatest restoration of dealkylation activity, 32% relative to a control not treated with EDTA, while Mn(II) addition results in the smallest restoration of dealkylation activity, only 3% relative to an untreated control. The structure and coordination of the zinc center has been investigated by X-ray absorption spectroscopy. EXAFS data are best interpreted by an average coordination of 2-3 histidine ligands and 1-2 non-histidine O/N ligands. Since catalytic zinc centers in other zinc metalloenzymes generally exhibit only O/N ligands to the zinc atom, a zinc-bound water or hydroxide may serve as a general base for the abstraction of the hydroxyl proton from the carbinolamide intermediate. Alternatively, the zinc may function in a structural role. PMID- 9405059 TI - Redox thermodynamics of the native and alkaline forms of eukaryotic and bacterial class I cytochromes c. AB - The reduction potentials of beef heart cytochrome c and cytochromes c2 from Rhodopseudomonas palustris, Rhodobacter sphaeroides, and Rhodobacter capsulatus were measured through direct electrochemistry at a surface-modified gold electrode as a function of temperature in nonisothermal experiments carried out at neutral and alkaline pH values. The thermodynamic parameters for protein reduction (DeltaS degrees rc and DeltaH degrees rc) were determined for the native and alkaline conformers. Enthalpy and entropy terms underlying species dependent differences in E degrees and pH- and temperature-induced E degrees changes for a given cytochrome were analyzed. The difference of about +0.1 V in E degrees between cytochromes c2 and the eukaryotic species can be separated into an enthalpic term (-DeltaDeltaH degrees rc/F) of +0.130 V and an entropic term (TDeltaDeltaS degrees rc/F) of -0.040 V. Hence, the higher potential of the bacterial species appears to be determined entirely by a greater enthalpic stabilization of the reduced state. Analogously, the much lower potential of the alkaline conformer(s) as compared to the native species is by far enthalpic in origin for both protein families, and is largely determined by the substitution of Met for Lys in axial heme ligation. Instead, the biphasic E degrees /temperature profile for the native cytochromes is due to a difference in reduction entropy between the conformers at low and high temperatures. Temperature-dependent 1H NMR experiments suggest that the temperature-induced transition also involves a change in orientation of the axial methionine ligand with respect to the heme plane. PMID- 9405060 TI - Fast cytochrome bo from Escherichia coli binds two molecules of nitric oxide at CuB. AB - The reaction of nitric oxide (NO) with fast cytochrome bo from Escherichia coli has been studied by electronic absorption, MCD, and EPR spectroscopy. Titration of the enzyme with NO showed the formation of two distinct species, consistent with NO binding stoichiometries of 1:1 and 2:1 with observed dissociation constants at pH 7.5 of approximately 2.3 x 10(-)6 and 3.3 x 10(-)5 M. Monitoring the titration by EPR spectroscopy revealed that the broad EPR signals at g approximately 7.3, 3.7, and 2.8 due to magnetic interaction between high-spin heme o (S = 5/2) and CuBII (S = 1/2) are lost. A high-spin heme o signal at g = 6.0 appears as the 1:1 complex is formed but is lost again on formation of the 2:1 complex, which is EPR silent. The absorption spectrum shows that heme o remains in the high-spin FeIII state throughout the titration. These results are consistent with the binding of up to two NO molecules at CuBII. This has been confirmed by studies with the Cl- adduct of fast cytochrome bo. MCD evidence shows that heme o remains ligated by histidine and water. Addition of excess NO to the Cl- adduct leads to the appearance of a high-spin FeIII heme EPR signal. Hence chloride ion binds to CuB, blocking the binding of a second NO molecule. These results suggest a mechanism for the reduction of NO to nitrous oxide by cytochrome bo and cytochrome c oxidase in which the binding of two cis NO molecules at CuB permits the formation of an N-N bond and the abstraction of oxygen by the heme group. PMID- 9405061 TI - Two enzymes with a common function but different heme ligands in the forms as isolated. Optical and magnetic properties of the heme groups in the oxidized forms of nitrite reductase, cytochrome cd1, from Pseudomonas stutzeri and Thiosphaera pantotropha. AB - It is shown that, in the oxidized state, heme c of Pseudomonas stutzeri (ZoBell strain) cytochrome cd1 has histidine-methionine ligation as observed for cytochrome cd1 from Pseudomonas aeruginosa [Sutherland, J., Greenwood, C., Peterson, J., and Thomson, A. J. (1986) Biochem. J. 233, 893-898]. However, the X ray structure of Thiosphaera pantotropha cytochrome cd1 reveals bis-histidine ligation for heme c. It is confirmed by EPR and near-infrared (NIR) MCD measurements that the bis-histidine coordination remains unaltered in the solution phase. Hence, the difference between the heme c ligation states defines two distinct classes of oxidized cytochromes cd1 as isolated. A weak feature in the T. pantotropha NIR MCD at 1900 nm suggests that a small population of heme c has histidine-methionine coordination. The ligation state of heme d1 cannot be defined with the same level of confidence, because the porphyrin-to-Fe(III) charge-transfer (CT) bands are less well characterized for this class of partially reduced porphyrin ring. However, variable temperature absorption and MCD spectra show that, in the T. pantotropha enzyme, heme d1 exists in a thermal low-spin/high-spin mixture with the low-spin as the ground state, whereas in P. stutzeri cytochrome cd1, and d1 heme is low-spin at all temperatures. A weak band, assigned as the heme d1 porphyrin-pi(a1u,a2u)-to-ferric(d) charge-transfer transition has been identified for the first time at 2170 nm. Its magnetic properties show the heme d1 to have an unusual (dxz,yz)4(dxy)1 electronic ground state as is found for low-spin Fe(III) chlorins [Cheesman, M. R., and Walker, F. A. (1996) J. Am. Chem. Soc. 118, 7373-7380]. It is proposed that the localization of the Fe(III) unpaired d-electron in an orbital lying in the heme plane may decrease the affinity of the Fe(III) heme for unsaturated ligands such as NO. Although heme d1 in the enzymes from P. stutzeri and T. pantotropha shows different temperature-dependent spin properties, the positions of the low-spin Fe(III) alpha-absorption band, at approximately 640 nm, are very similar to those observed for cytochromes cd1 from eight other sources, suggesting that all have similar strength fields from the axial ligands and, hence, that all have the same coordination, namely histidine-tyrosine or possibly histidine-hydroxide at the heme. PMID- 9405062 TI - Bicarbonate may Be required for ligation of manganese in the oxygen-evolving complex of photosystem II. AB - It was previously shown in the photosystem II membrane preparation DT-20 that photoxidation of the oxygen-evolving manganese cluster was blocked by 0.1 mM formate, unless 0.2 mM bicarbonate was present as well [Wincencjusz, H., Allakhverdiev, S. I., Klimov, V. V., and Van Gorkom, H. J. (1996) Biochim. Biophys. Acta 1273, 1-3]. Here it is shown by measurements of EPR signal II that oxidation of the secondary electron donor, YZ, is not inhibited. However, the reduction of is greatly slowed and occurs largely by back reaction with reduced acceptors. Bicarbonate is shown to prevent the loss of fast electron donation to . The release of about one or two free Mn2+ per photosystem II during formate treatment, and the fact that these effects are mimicked by Mn-depletion, suggests that formate may act by replacing a bicarbonate which is essential for Mn binding. Irreversible light-induced rebinding in an EPR-silent form of Mn2+ that was added to Mn-depleted DT-20 was indeed found to depend on the presence of bicarbonate, as did the reconstitution in such material of both the fast electron donation to and the UV absorbance changes characteristic of a functional oxygen evolving complex. It is concluded that bicarbonate may be an essential ligand of the functional Mn cluster. PMID- 9405063 TI - Influence of the protein binding site on the absorption properties of the monomeric bacteriochlorophyll in Rhodobacter sphaeroides LH2 complex. AB - Resonance Raman spectroscopy was performed on peripheral light-harvesting proteins from Rhodobacter sphaeroides in which the residue betaArg-10 has been modified by site-selected mutagenesis. We show that this residue is indeed involved (as proposed by X-ray crystallographic studies on the LH2 complex from Rhodopseudomonas acidophila), in an H-bond with the acetyl carbonyl of the 800 nm absorbing BChl in these proteins (B800), and that the presence of such an H-bond induces a ca. 10 nm red shift of the lowest energy transition (Qy) of this molecule. Moreover, other parameters involved in the fine tuning of the absorption of the B800 molecules may be determined from our experiments, and we propose that the local electromagnetic properties of the B800 binding site may induce an additional 10 nm red shift of this transition. These results constitute the first experimental evidence for the parameters able to modify in vivo the absorption of "monomeric" BChl molecules, i.e. BChl not involved in strong excitonic interactions, and will be of great help for understanding the absorption properties of such pigments in other light-harvesting systems. PMID- 9405064 TI - Reconstitution of bovine A1 adenosine receptors and G proteins in phospholipid vesicles: betagamma-subunit composition influences guanine nucleotide exchange and agonist binding. AB - We have studied the interactions of purified A1 adenosine receptors and G proteins reconstituted into phospholipid vesicles to investigate how the betagamma composition of G protein heterotrimers influences coupling. Recombinant hexahistidine-tagged bovine A1 adenosine receptors were expressed in Sf9 cells and purified to homogeneity by sequential chromatography over heparin-sepharose, xanthine amino congener-agarose, and nickel-nitrilotriacetic acid columns. These receptors were reconstituted with pure recombinant G proteins of defined subunit composition. Receptor-G protein complexes containing alphai2 and beta1gamma2 or beta1gamma3 and stimulated with the agonist, (R)-phenylisopropyladenosine, exchange guanine nucleotide 2-3 times more rapidly than do complexes containing beta1gamma1. This difference is not overcome by increasing the concentration of betagamma subunits. Receptor-G protein complexes containing beta1gamma1 also bind less of the agonist, [125I]-iodoaminobenzyladenosine (125I-ABA), than do complexes containing beta1gamma3. Kinetic experiments show that 125I-ABA dissociates 2-fold more rapidly from receptor-G protein complexes containing beta1gamma1 than from complexes containing the other betagamma subunits. The affinity of the interaction between immobilized Galphai2 subunits and beta1gamma1 or beta1gamma2 measured with an optical biosensor in the absence of receptor is similar. Taken together, these data implicate the gamma-subunit in influencing the interaction between the A1 adenosine receptor and G proteins. PMID- 9405065 TI - Use of Trp mutations to evaluate the conformational behavior and membrane insertion of A and B chains in whole diphtheria toxin. AB - The structure of diphtheria toxin was examined using its Trp fluorescence. To examine the interactions of the A and B chains of the toxin independently, mutants were constructed in which Trp residues were restricted to either the A or the B chain. The conformation and stability of the mutants were very similar to those of the wild-type protein. In addition, they underwent the low-pH conformational transition and membrane insertion at about the same pH as wild type toxin. This shows Trp do not play a critical role in these processes which are necessary for the translocation of toxin across endosomal membranes in vivo. There was a shift in fluorescence of the Trp mutants which showed the low-pH induced transition increases exposure of both the A and B Trp to a more polar environment. This supports a model in which the interdomain interactions present at neutral pH break down at low pH. To evaluate the location of the A and B chains in the membrane, the fluorescence quenching of model membrane inserted toxin was measured. Comparison of the amount of quenching by lipid labeled with nitroxides localized at shallow, medium, or deep depths within the bilayer demonstrated that both the A and B chains insert deeply, but the A chain Trp are somewhat less deeply inserted. Trp on the A chain are also less exposed to lipid than on the B chain, as judged by their weaker quenching by the nitroxide-labeled lipid. This conclusion was supported by the observation that the Trp of membrane inserted isolated A chain is more lipid-exposed than when the A chain is part of the whole toxin. Both the A and B chain Trp become less exposed to lipid after neutralizing pH. However, both chains remain inserted, with at least part of the B chain remaining deeply inserted. These results support the "partial wrapper" model in which both the A and B chains are inserted but contacts between the two chains significantly reduce the exposure of the A chain to lipid. PMID- 9405066 TI - Role of prohormone convertases in pro-neuropeptide Y processing: coexpression and in vitro kinetic investigations. AB - Proneuropeptide Y (ProNPY) undergoes cleavage at a single dibasic site Lys38 Arg39 resulting in the formation of 1-39 amino acid NPY which is further processed successively by carboxypeptidase-like and peptidylglycine alpha amidating monooxygenase enzymes. To investigate whether prohormone convertases are involved in ProNPY processing, a vaccinia virus derived expression system was used to coexpress recombinant ProNPY with each of the prohormone convertases PC1/3, PC2, furin, and PACE4 in Neuro2A and NIH 3T3 cell lines as regulated neuroendocrine and constitutive prototype cell lines, respectively. The analysis of processed products shows that only PC1/3 generates NPY in NIH 3T3 cells while both PC1/3 and PC2 are able to generate NPY in Neuro2A cells. The convertases furin and PACE4 are unable to process ProNPY in either cell line. Moreover, comparative in vitro cleavage of recombinant NPY precursor by the enzymes PC1/3, PC2 and furin shows that only PC1/3 and PC2 are involved in specific cleavage of the dibasic site. Kinetic studies demonstrate that PC1/3 cleaves ProNPY more efficiently than PC2. The main difference between the cleavage efficiency is observed in the Vmax values whereas no major difference is observed in Km values. In addition the cleavage by PC1/3 and PC2 of two peptides reproducing the dibasic cleavage site with different amino acid sequence lengths namely (20-49)-ProNPY and (28-43)-ProNPY was studied. These shortened ProNPY substrates, when recognized by the enzymes, are more efficiently cleaved than ProNPY itself. The shortest peptide is not cleaved by PC2 while it is by PC1/3. On the basis of these observations it is proposed, first, that the constitutive secreted NPY does not result from the cleavage carried out by ubiquitously expressed enzymes furin and PACE4; second, that PC1/3 and PC2 are not equipotent in the cleavage of ProNPY; and third, substrate peptide length might discriminate PC1/3 and PC2 processing activity. PMID- 9405067 TI - Effect of Mts1 on the structure and activity of nonmuscle myosin II. AB - The mts1 gene codes for a 9 kDa protein belonging to the S100 subfamily of Ca2+ binding proteins and is known to play a role in metastasis. Its role in metastasis may be through cellular locomotion, as transfection of mts1 into mouse mammary adenocarcinoma cells increases cellular motility in modified Boyden chemotaxis chambers. The Mts1 protein interacts with nonmuscle myosin II in the presence of Ca2+ with an affinity of approximately 7.9 x 10(4) M-1 and an approximate stoichiometry of 3 mol of Mts1/mol of myosin heavy chain. No interaction was found with myosin I or myosin V. The binding site of Mts1 on myosin is in the rod region, particularly to the light meromyosin portion of the rod. To understand the mechanism by which Mts1 alters cellular motility, we examined its effect on myosin structure and activity. Cosedimentation analysis and electron microscopy suggest that Mts1 destabilizes myosin filaments. In the presence of Ca2+, Mts1 inhibits the actin-activated MgATPase activity of myosin in vitro. The data demonstrate an effect of Mts1 on both myosin structure and function, and suggest a route through which Mts1 affects motility as well as metastasis. PMID- 9405068 TI - Neuromedin B receptor activation causes tyrosine phosphorylation of p125FAK by a phospholipase C independent mechanism which requires p21rho and integrity of the actin cytoskeleton. AB - Recent studies show that tyrosine phosphorylation by a number of neuropeptides may be an important intracellular pathway in mediating changes in cell function, particularly related to growth. Neuromedin B (NMB), a mammalian bombesin related peptide, functions through a distinct receptor, the neuromedin B receptor (NMB R), of which little is known about its cellular basis of action. In the present study we explored the ability of NMB-R activation to cause tyrosine phosphorylation of focal adhesion kinase (p125(FAK)), an important substrate for tyrosine phosphorylation by other neuropeptides. NMB caused rapid increases in p125(FAK) phosphorylation which reached maximum at 2 min in both rat C6 glioblastoma cells which possess native NMB-Rs and rat neuromedin B receptor (rNMR-R) transfected BALB 3T3 cells. NMB had a half-maximal effect was at 0.4 nM and was 30-fold more potent than gastrin-releasing peptide (GRP). The stoichiometric relationships between increased p125(FAK) tyrosine phosphorylation and other cellular processes was similar in both C6 cells and rNMB-R transfected cells. TPA (1 microM) caused 45% and the calcium ionophore, A23187, 11% of maximal tyrosine phosphorylation of p125(FAK) seen with NMB. A23187 potentiated the effect of TPA. Pretreatment with the selective PKC inhibitor, GF109203X, inhibited TPA-induced p125(FAK) tyrosine phosphorylation, but it had no effect on the NMB stimulation. Pretreatment with thapsigargin completely inhibited NMB stimulated increases in [Ca2+]i, but had no effect on NMB-stimulation of p125(FAK) phosphorylation either alone or with GF109203X. The tyrosine kinase inhibitor, tyrphostin A25, inhibited NMB-induced phosphorylation of p125(FAK) by 52%. However, tyrphostin A25 did not inhibit NMB-stimulated increases in [3H]inositol phosphates. Cytochalasin D, an agent which disrupts actin microfilaments, inhibited BN- and TPA-induced tyrosine phosphorylation of p125(FAK) completely. In contrast, colchicine, an agent which disrupts microtubules, had no effect. Pretreatment with Clostridium botulinum C3 exoenzyme which inactivates the small GTP-binding protein rho p21, also inhibited tyrosine phosphorylation of p125(FAK) by 55%. These results demonstrate that activation of NMB-R can cause rapid tyrosine phosphorylation of p125(FAK). NMB-induced tyrosine phosphorylation of p125(FAK) is independent of NMB-induced changes in [Ca2+]i or PKC. The integrity of the actin cytoskeleton but not of microtubules is necessary for NMB-stimulated phosphorylation of p125(FAK). The ras-related small GTP binding protein rho p21 is at least partially involved in mediating NMB-induced tyrosine phosphorylation of p125(FAK). These results suggest that similar to some other neuropeptides, activation of this pathway may be an important mechanism in mediating cellular changes by this receptor such as growth. PMID- 9405069 TI - Characterization of the interthiol acyltransferase reaction catalyzed by the beta ketoacyl synthase domain of the animal fatty acid synthase. AB - The enzyme activity responsible for translocation of saturated acyl chains from the 4'-phosphopantetheine of the acyl carrier protein to the active site cysteine of the beta-ketoacyl synthase in the animal fatty acid synthase has been identified. An enzyme assay was devised that allows uncoupling of the interthiol transfer step from the condensation reaction. Experiments with various fatty acid synthase mutants indicate clearly that catalysis of the transfer of saturated acyl moieties from the 4'-phosphopantetheine thiol to the active site cysteine thiol, Cys-161, is an inherent property of the beta-ketoacyl synthase domain. Catalytic efficiency of the interthiol transferase increases from C2 to C12 and decreases with increasing chain-lengths beyond C12. Malonyl, beta-hydroxybutyryl, and crotonyl thioesters are not substrates for the transferase, whereas the beta ketobutyryl moiety is a poor substrate. These features of the substrate specificity are exactly as predicted for a transferase that fulfills the proposed role in the fatty acid synthase reaction sequence and indicate that this activity plays an important role in determining the overall specificity of the beta ketoacyl synthase reaction. PMID- 9405093 TI - Xenopus hindbrain patterning requires retinoid signaling. AB - We have asked how posterior neural tissue is patterned in Xenopus by assaying the involvement of endogenous retinoic acid (RA) in this process and by using the labial Hox gene, HoxD1, as a posterior marker. Although RA is able to inhibit anterior gene expression and activate expression of more posterior genes, the normal role of retinoids in anteroposterior (A/P) patterning is unclear. HoxD1 is an early posterior neurectodermal marker, expressed from midgastrula with a later anterior expression limit in the future hindbrain. We previously showed that HoxD1 was induced as an immediate early response to retinoic acid in naive ectoderm (animal caps). Here, we use a truncated RARalpha2.2 receptor (RARDelta) to dominantly interfere with retinoid signaling. In embryos injected with RARDelta expression of HoxD1 is eliminated. Conjugates of ectoderm and dorsolateral mesoderm show that retinoid receptors are required in the ectoderm for HoxD1 induction. Further, expression of Krox-20 in r3 and r5 of the presumptive hindbrain is compressed into a single stripe that suggests elimination of r5. RARalpha2.2 expression almost precisely overlaps that of HoxD1, suggesting that this receptor may normally activate HoxD1. Expression of neither more anterior genes including cement gland, forebrain, and midbrain markers nor a more posterior spinal cord marker is affected by RARDelta. These data suggest that the posterior hindbrain is the region of the nervous system most sensitive to retinoid loss. Finally, we compare the ability of RA and fibroblast growth factor (FGF) to posteriorize isolated anterior neurectoderm and show that both factors can act directly on this substrate. RA acts in a more anterior domain than does FGF; however, neither factor is equivalent to the natural posteriorizing capacity of the posterior mesoderm. We propose that endogenous retinoid and FGF signals pattern largely nonoverlapping regions along the A/P axis and that posterior neural patterning requires multiple inducers. PMID- 9405070 TI - Kinetic and secondary structure analysis of Naegleria andersoni GIR1, a group I ribozyme whose putative biological function is site-specific hydrolysis. AB - NanGIR1 is a catalytic element inserted in the P6 loop of a group I intron (NanGIR2) in the small subunit rRNA precursor of the protist Naegleria andersoni [Einvik, C., Decatur, W. A., Embley, T. M., Vogt, V. M., and Johansen, S. (1997) RNA 3, 710-720]. It catalyzes site-specific hydrolysis at an internal processing site (IPS) after a G residue that immediately follows the P9 stem-loop. Functional and structural analyses were initiated to compare NanGIR1 to group I introns that carry out self-splicing. Chemical modification and site-directed mutagenesis studies showed that NanGIR1 shares many structural elements with other group I introns, but also contains a pseudoknot (P15), which is important for catalytic activity. Deletion analysis revealed the boundaries of the minimum self-cleaving unit (178 nucleotides). The rate of self-cleavage was measured as a function of mono- and divalent ion concentration, temperature, and pH. The reaction at the IPS yields 5'-phosphate and 3'-hydroxyl termini, requires Mg2+or Mn2+ ions, and is first-order in [OH-] between pH 5.0 and 8.5. The latter results suggest that the nucleophile in the reaction is hydroxide or possibly a Mg2+ coordinated hydroxide. With a second-order rate constant of 1 x 10(5) min-1 M-1, the self-cleavage reaction of NanGIR1 is 2 orders of magnitude faster than a similar site-specific hydrolysis reaction of the circular form of the Tetrahymena group I intron. PMID- 9405095 TI - WNT-7a induces axonal remodeling and increases synapsin I levels in cerebellar neurons. AB - WNT factors play a key role in early patterning of the embryo. However, expression of Wnt genes after cell commitment suggests additional roles in later developmental processes. We report here that Wnt-7a is expressed in cerebellar granule cell neurons as they begin to extend processes and form synapses. WNT-7a increases axonal spreading and branching in cultured granule cells. Moreover, WNT 7a increases the levels of synapsin I, a presynaptic protein involved in synapse formation and function. Lithium mimics WNT-7a in granule cells by inhibiting GSK 3beta, a component of the WNT signaling pathway. These results suggest a direct effect of WNT-7a in the regulation of neuronal cytoskeleton and synapsin I in granule cell neurons. We propose that WNT proteins have a novel function in the formation of neuronal connections. PMID- 9405094 TI - TAL1/SCL is expressed in endothelial progenitor cells/angioblasts and defines a dorsal-to-ventral gradient of vasculogenesis. AB - In this study we establish that TAL1/SCL, a member of the helix-loop-helix family of transcription factors, and an important regulator of the hematopoietic lineage in mice, is expressed in the endothelial lineage of avians. The earliest events of vascular development were examined using antibodies to TAL1/SCL, and the QH1 antibody, an established marker of quail endothelial cells. Analyses using double immunofluorescence confocal microscopy show that: (i) TAL1/SCL is expressed by both quail and chicken endothelial cells; (ii) TAL1/SCL expression precedes that of the QH1 epitope; and (iii) TAL1/SCL, but not QH1, expression defines a subpopulation of primordial cells within the splanchnic mesoderm. Collectively these data suggest that TAL1/SCL-positive/QH1-negative cells are angioblasts. Further, using TAL1/SCL expression as a marker of the endothelial lineage, we demonstrate that in addition to the previously described cranial-to-caudal gradient, there is a dorsal-to-ventral progression of vasculogenesis. PMID- 9405096 TI - Expression of T protein in the primitive streak is necessary and sufficient for posterior mesoderm movement and somite differentiation. AB - A characteristic abnormality of chimeras composed of wildtype and T/T (Brachyury) mutant embryonic stem cells is the aggregation and accumulation of mutant cells in the primitive streak and its descendant, the tail bud (V. Wilson, L. Manson, W. C. Skarnes, and R. S. P. Beddington (1995). Development 121, 877-886). To demonstrate that this aberrant behaviour of mutant cells in the streak is due only to the absence of wild-type T protein and to investigate dosage effects of T function on cell deployment during gastrulation, a vector expressing T under the control of its own promoter (which results in T expression in the primitive streak but not in the notochord) was introduced into T/T mutant ES cells carrying a ubiquitous lacZ lineage marker. Four clones (TR clones) that express T appropriately in the streak and rescue abnormal chimeric morphology were recovered. In chimeras, these four clones fall into two distinct categories with respect to their ability to exit from the primitive streak and their subsequent tissue colonisation profile. TR1 and TR4 descendants no longer accumulated in the tail bud and gave rise to all types of mesoderm as well as colonising ventral neurectoderm. Interestingly, TR2 and TR5 cells (which express higher levels of T protein than TR1 and TR4 in vitro) tended to exit the streak prematurely, showed a marked reduction in posterior mesoderm colonisation, and were virtually excluded from ventral neurectoderm. However, while descendants of all four TR clones can colonise dermomyotome at all axial levels, the parent T/T mutant cells only contribute to this tissue rostral to the forelimb bud and are completely excluded from more caudal dermomyotome. These results show that the abnormal aggregation of mutant cells homozygous for the Brachyury deletion (approximately 200 kb) can be ascribed solely to the lack of wild-type T protein, as can the failure of T/T cells to colonise caudal dermomyotome. They also suggest that patterns of cell recruitment from the streak can be influenced by the level of T expression. PMID- 9405097 TI - The POU gene ceh-18 promotes gonadal sheath cell differentiation and function required for meiotic maturation and ovulation in Caenorhabditis elegans. AB - In Caenorhabditis elegans, specialized contractile myoepithelial cells of the somatic gonad, the gonadal sheath cells, are closely apposed to oocytes and are required for normal meiotic maturation and ovulation. Previously we found that mutations in the ceh-18 gene, which encodes a POU-class homeoprotein expressed in sheath cells, result in oocyte defects. To determine the basis for these oocyte defects, we have used time-lapse video Nomarski microscopy to observe meiotic maturation, ovulation, and early embryogenesis in ceh-18 mutants. In ceh-18 mutants sheath cell contractions are weaker, less frequent, and uncoordinated throughout the sequence of ovulation events, and ovulation is defective. Defective ovulation can result in the formation of endomitotic oocytes in the gonad, the formation of haploid embryos, and reversals in embryonic polarity. ceh 18 mutant oocytes exhibit defects prior to nuclear envelope breakdown, suggesting that they are physiologically different from the wild type. We observed delays in meiotic maturation, as well as maturation out of the normal spatial and temporal sequence, suggesting that proximal sheath cells directly or indirectly promote and spatially restrict meiotic maturation. Analysis of sheath cell differentiation in ceh-18 mutants using antibodies to proteins of the contractile apparatus reveals that although contractile proteins are expressed, the sheath cells appear disorganized. Transmission electron microscopy reveals that ceh-18 mutant sheath cells are morphologically irregular and only loosely cover oocytes. Taken together, these observations indicate that ceh-18 is a crucial determinant of sheath cell differentiation, a function required for normal meiotic maturation and ovulation. PMID- 9405098 TI - Localized calcium transients accompany furrow positioning, propagation, and deepening during the early cleavage period of zebrafish embryos. AB - Through the injection of f-aequorin (a calcium-specific luminescent reporter) and the use of an imaging photon detector, we see a distinct localized elevation of intracellular calcium that accompanies the appearance of the first furrow arc at the blastodisc surface: the furrow positioning signal. As the leading edges of the arc progress outward toward the margins of the blastodisc, they are accompanied by two subsurface slow calcium waves moving at about 0.2 micron/s: the furrow propagation signal. As these wave fronts approach the edge of the blastodisc, another calcium signal arises in the central region where the positioning signal originally appeared. Like the propagation signal, it extends outward to the margins of the blastodisc, but in this case it also moves downward, accompanying the deepening process that separates the daughter cells: the furrow deepening signal. Both of these furrow deepening progressions move at around 0.1 to 0.2 micron/s. The deepening signal begins to diminish from the center outward, returning to precleavage resting levels on completion of cytokinesis. The signaling sequence is repeated during the second cell division cycle. These localized transients do not require external calcium and they can be dissipated after they have begun by introducing calcium shuttle buffers, resulting in furrow delocalization and regression. They also occur in parthenogenetically activated eggs in which, in an attenuated form, they accompany abortive cleavages. PMID- 9405099 TI - P19 embryonal carcinoma cells: a model system for studying neural tube induction of skeletal myogenesis. AB - A model experimental system for investigating myogenic induction signals has been devised with mouse P19 embryonal carcinoma cells. When cocultured with pieces of chick neural tube, aggregated P19 cells are induced to become skeletal muscle. The most potent inducing activity is localized to the dorsal neural tube. Less activity was found in the ventral neural tube, notochord, ectoderm, and lateral plate mesoderm, and none was detected in the neural retina. These results suggest that P19 cells may be a useful model system for investigating the mechanisms underlying induction of somite myogenesis. PMID- 9405100 TI - Activation of the receptor tyrosine kinase Kit is required for the proliferation of melanoblasts in the mouse embryo. AB - The development of neural crest-derived melanocytes, as well as haematopoietic and germ cells, is affected by mutations of the Kit and Mgf genes, which lead to dominant spotting (W) or steel (Sl) phenotypes. Mgf codes for the ligand of the receptor tyrosine kinase encoded by the Kit locus. KitW-v, a point mutation exerting a dominant negative effect, causes a substantial reduction in tyrosine kinase activity of the Kit receptor and leads to a characteristic pigmentation phenotype, namely dilute coat colour and a white ventral and head spot with reduced pigmentation of the feet and tail in the heterozygous animal, as well as slight anaemia. Homozygous animals lack coat pigmentation and are severely anaemic and infertile. Dct is a marker for cells of the melanoblast lineage. In order to study these cells in detail we have generated transgenic mouse lines carrying the lacZ reporter under the control of the Dct promoter and have used the embryonic expression of the reporter to identify early melanoblasts before they begin to produce pigment. Our transgenic lines have simplified the study of melanoblasts in the mouse embryo, and by crossing our mice with KitW-v mutants we have been able to identify the midgestation stages at which melanoblasts rely critically on Mgf/Kit interactions. We conclude that the survival of immature melanoblasts depends crucially upon Kit signalling up until E11, and later in development Kit plays a vital role in melanoblast proliferation. Our data do not describe a dependence upon Kit for melanoblast migration or differentiation. PMID- 9405101 TI - Isolation and characterization of chondroitin sulfate proteoglycans from embryonic quail that influence neural crest cell behavior. AB - The movement of neural crest cells is controlled in part by extracellular matrix. Aggrecan, the chondroitin sulfate proteoglycan from adult cartilage, curtails the ability of neural crest cells to adhere, spread, and move across otherwise favorable matrix substrates in vitro. Our aim was to isolate, characterize, and compare the structure and effect on neural crest cells of aggrecan and proteoglycans purified from the tissues through which neural crest cells migrate. We metabolically radiolabeled proteoglycans in E2.5 quail embryos and isolated and characterized proteoglycans from E3.3 quail trunk and limb bud. The major labeled proteoglycan was highly negatively charged, similar in hydrodynamic size to chick limb bud versican/PG-M, smaller than adult cartilage aggrecan but larger than reported for embryonic sternal cartilage aggrecan. The molecular weight of the iodinated core protein was about 400 kDa, which is more than reported for aggrecan but less than that of chick versican/PG-M. The proteoglycan bore chondroitin sulfate glycosaminoglycan chains of 45 kDa, which is larger than those of aggrecan. It lacked dermatan sulfate, heparan sulfate, or keratan sulfate chains. It bound to collagen type I, like aggrecan, but not to fibronectin (unlike versican/PG-M), collagen type IV, or laminin-1 in solid-phase assays and it bound to hyaluronate in gel-shift assays. When added at concentrations between 10 and 30 microg/ml to substrates of fibronectin, trunk proteoglycan inhibited neural crest cell spreading and migration. Attenuation of cell spreading was shown to be the most sensitive and titratable measure of the effect on neural crest cells. This effect was sensitive to digestion with chondroitinase ABC. Similar cell behavior was also produced by aggrecan and the small dermatan sulfate proteoglycan decorin; however, 30-fold more aggrecan was required to produce an effect of similar magnitude. When added in solution to neural crest cells which were already spread and migrating on fibronectin, the embryonic proteoglycan rapidly and reversibly caused complete rounding of the cells, being at least 30-fold more potent than aggrecan in this activity. PMID- 9405102 TI - The fucose sulfate polymer of egg jelly binds to sperm REJ and is the inducer of the sea urchin sperm acrosome reaction. AB - The sea urchin sperm cell is an advantageous model for studying ligand-mediated exocytosis. Sperm can be obtained in vast quantities and induced to undergo exocytosis of the acrosomal vesicle with great synchrony. During sea urchin fertilization, egg jelly (EJ) triggers the sperm acrosome reaction (AR) which is required for sperm binding and fusion with the egg. Uncertainty exists as to the exact biochemical nature of the AR inducer. The following study was performed in an attempt to clarify the nature of the inducer. EJ from individual females (Strongylocentrotus purpuratus) was analyzed on SDS-PAGE gels. Each female had a unique composition of EJ macromolecules, but all females possessed the previously described fucose sulfate polymer (FSP). Two electrophoretic isotypes of FSP were discovered; 87% of females had only one isotype and 13% had both. Both FSP isotypes bound to the REJ protein (receptor for egg jelly) purified from sperm. The two FSP isotypes had almost equal potency in inducing the AR. EJ was fractionated by DEAE chromatography in 6 M urea/4% beta-mercaptoethanol. All AR inducing activity coeluted with FSP. FSP, purified by trypsin digestion followed by dialysis, was twice as active as the non-trypsin-digested control at inducing the AR. EJ was digested with proteinase K, boiled in detergent and beta mercaptoethanol, and subjected to sucrose density gradient sedimentation. The FSP and AR activity had superimposable sedimentation patterns. Purified FSP had no associated peptide component. Sperm from individual males differed in the concentration dependency of purified FSP to induce the AR. The data indicate that the 138/82 kDa EJ glycoproteins, previously thought to act as AR inducers, do not appear to be involved in triggering the AR. The data are consistent with the hypothesis that FSP is the only inducer of the AR of this sea urchin species. PMID- 9405103 TI - Cell adhesion molecules regulate guidance of dorsal root ganglion axons in the marginal zone and their invasion into the mantle layer of embryonic spinal cord. AB - In order to elucidate the mechanisms regulating the projections of dorsal root ganglion (DRG) axons in the dorsal funiculus and invasion into target regions in the mantle layer (prospective gray matter) of the spinal cord, we examined the interactions between DRG axons and spinal cord. DRG neurons were dissociated from chick embryos and cultured for 1-2 days on cryostat sections of the spinal cord at embryonic day 5 (E5) or at E9. E5 and E9 DRG neurons extended neurites onto both marginal zone (prospective white matter) and mantle layer (prospective gray matter) of the spinal cord, suggesting that both of these regions are permissive for neurite growth. When E5 DRG neurites approached cryosections of E5 spinal cord from outside, most of them ran in the marginal zone without invading the mantle layer. In contrast, about half of E9 DRG neurites entered the mantle layer after crossing the marginal zone of E9 spinal cord. These growth patterns of DRG neurites on spinal marginal zone and mantle layer are similar to the pathway formation of DRG axons at comparable stages in vivo; DRG axons run exclusively in the prospective dorsal funiculus before E6, and enter the mantle layer (prospective dorsal horn) to reach the target regions by E9. Perturbation of functions of Ng-CAM, Nr-CAM, and axonin-1/SC2 by adding the specific antibodies in the culture medium increased the ratio of DRG neurites entering the mantle layer of E5 spinal cord, suggesting that these cell adhesion molecules are involved in keeping DRG neurites in the marginal zone. Taken together with the expression of Ng-CAM, Nr-CAM, and axonin-1/SC2, these CAMs on DRG axons may regulate the guidance of these axons in the marginal zone before E6, and the subsequent decrease in the relative levels of these CAMs might allow DRG axons to invade the target mantle layer. PMID- 9405104 TI - Anteroposterior gradient of epithelial transformation during amphibian intestinal remodeling: immunohistochemical detection of intestinal fatty acid-binding protein. AB - To determine whether the remodeling of the well-organized intestinal epithelium during amphibian metamorphosis is regionally regulated along the anteroposterior axis of the intestine, we raised a polyclonal antibody against the Xenopus laevis intestinal fatty acid-binding protein (IFABP), which is known to be specifically expressed in intestinal absorptive cells, and examined immunohistochemically the differentiation, proliferation, and apoptosis of the epithelial cells throughout X. laevis small intestine. During pre- and prometamorphosis, IFABP-immunoreactive (ir) epithelial cells were localized only in the anterior half of the larval intestine. At the beginning of metamorphic climax, apoptotic cells detected by nick end-labeling (TUNEL) suddenly increased in number in the entire larval epithelium, concurrently with the appearance of adult epithelial primordia. Subsequently, the adult primordia in the anterior part of the intestine developed more rapidly by active cell proliferation than those in the posterior part, and replaced the larval epithelial cells earlier than those in the posterior part. IFABP-ir cells in the adult epithelium were first detectable at the tips of newly formed folds in the proximal part of the intestine. Thereafter, IFABP expression gradually progressed both in the anteroposterior direction and in the crest trough direction of the folds. These results suggest that developmental processes of the adult epithelium in the X. laevis intestine are regionally regulated along the anteroposterior axis of the intestine, which is maintained throughout metamorphosis, and along the trough-crest axis of the epithelial folds, which is newly established during metamorphosis. Furthermore, the regional differences in IFABP expression along the anteroposterior axis of the intestine were reproduced in organ cultures in vitro. In addition, IFABP expression was first down regulated and then reactivated in vitro when the anterior part, but not the posterior part, of the larval intestine was treated with thyroid hormone (TH) for extended periods. Therefore, it seems that, in addition to TH, an endogenous factor(s) localized in the intestine itself with an anteroposterior gradient participates in the development of the adult epithelium during amphibian metamorphosis. PMID- 9405105 TI - The homeobox gene PV.1 mediates specification of the prospective neural ectoderm in Xenopus embryos. AB - Bone morphogenetic protein 4 (BMP4), a member of the TGF beta superfamily, has been implicated in the dorsoventral specification of both mesoderm and ectoderm. High levels of BMP4 signaling appear to specify ventral lineages, while lower levels are causally associated with the development of dorsal lineages. We have previously identified a homeobox-containing transcription factor (PV. 1) which is a likely mediator of the ventralizing effects of BMP4 in the mesoderm. Here we provide evidence that PV.1 also functions downstream of BMP4 in the patterning of ectoderm, specifying epidermal and suppressing neural gene expression. PV.1 is expressed in the prospective neuroectoderm at the time of ectodermal fate determination. BMP4 and xSmad1 (a downstream effector of BMP4) induce PV.1 in uncommitted ectoderm and the dominant negative form of the BMP4 receptor (DN-BR) blocks PV.1 expression. In animal pole explants PV.1 counteracts the neuralizing effects of chordin and the DN-BR and restores them to their original epidermal fate. To address the physiological significance of these observations we employed an animal cap transplantation system and demonstrated that overexpression of PV.1 in the prospective neuroectoderm specifically blocks neurogenesis in intact embryos. Thus, PV.1 plays an important role in the ventralization of both mesoderm and ectoderm. We have previously shown that PV.1 is also preferentially expressed in the ventral endoderm, suggesting that this transcription factor may be involved in the ventralization of all three germ layers. PMID- 9405106 TI - Faithful expression of the Myf-5 gene during mouse myogenesis requires distant control regions: a transgene approach using yeast artificial chromosomes. AB - Myf-5, a member of the family of four muscle-specific basic helix-loop-helix (bHLH) transcription factors is the first to be expressed in somites, branchial arches, and limb buds during prenatal mouse development. However, little is known about control mechanisms which actually regulate Myf-5 gene activity within these various muscle-forming domains. To identify control regions that contribute to the correct spatiotemporal activity pattern of the Myf-5 gene during mouse embryogenesis, here we report the characterization of yeast artificial chromosomes (YACs) which faithfully direct muscle-specific expression of the gene in chimeric mouse embryos. Forty-five kilobases of sequence 5' to the Myf-5 gene together with 500 kb of 3' flanking DNA drives the correct Myf-5 expression in the mesenchyme of the visceral arches and in somites but not in the hypaxial muscles of limb buds. An additional 50 kb of DNA at the 5' end is required to activate Myf-5 gene expression in developing limbs. These results demonstrate for the first time that unexpectedly distant regions of the Myf-5 gene are necessary to recapitulate its precise developmental expression pattern. We also show that Myf-5 expression in hypaxial muscles and in somites and visceral arches is regulated by separate and distinct far upstream regions. The identification of these remote regulatory elements on YACs carrying the mouse Myf-5 gene constitutes the first important step toward further dissection of the complex mechanisms by which cell-autonomous and external cues control Myf-5 expression during skeletal muscle formation in the mouse embryo. PMID- 9405107 TI - Null mutations of the Dictyostelium cyclic nucleotide phosphodiesterase gene block chemotactic cell movement in developing aggregates. AB - Extracellular cAMP is a critical messenger in the multicellular development of the cellular slime mold Dictyostelium discoideum. The levels of cAMP are controlled by a cyclic nucleotide phosphodiesterase (PDE) that is secreted by the cells. The PDE gene (pdsA) is controlled by three promoters that permit expression during vegetative growth, during aggregation, and in prestalk cells of the older structures. Targeted disruption of the gene aborts development, and complementation with a modified pdsA restores development. Two distinct promoters must be used for full complementation, and an inhibitory domain of the PDE must be removed. We took advantage of newly isolated PDE-null cells and the natural chimerism of the organism to ask whether the absence of PDE affected individual cell behavior. PDE-null cells aggregated with isogenic wild-type cells in chimeric mixtures, but could not move in a coordinated manner in mounds. The wild type cells move inward toward the center of the mound, leaving many of the PDE null cells at the periphery of the aggregate. During the later stages of development, PDE-null cells in the chimera segregate to regions which correspond to the prestalk region and the rear of the slug. Participation in the prespore/spore population returns with the restoration of a modified pdsA to the null cells. PMID- 9405109 TI - EDITORIAL PMID- 9405108 TI - GDNF induces branching and increased cell proliferation in the ureter of the mouse. AB - The secreted signaling molecule GDNF is expressed in the metanephric mesenchyme and has recently been implicated as a factor necessary for development of the metanephric kidney. We have examined the effects of GDNF on mouse kidney explants. We show that GDNF increases cell proliferation in ureter tips. There is an increase in the number of ureter tips and expansion and fusion of adjacent tips and some tips appear to grow toward the source of GDNF. These events are accompanied by transcriptional upregulation of several genes localized to the tips, including its own receptor, c-ret, the transcription factor Sox9, and the signal Wnt-11. These results support a model in which GDNF supplied by the mesenchyme regulates growth and branching in the metanephric kidney through the local regulation of ureter tip-specific factors. PMID- 9405110 TI - Immunocytochemical and histological differences in the interrenal axis of feral brown trout, Salmo trutta, in metal-contaminated waters. AB - There are more CRH-like immunoreactive neurons in the preoptic nucleus and nucleus lateralis tuberis in the brain of feral brown trout, Salmo trutta, living in cadmium- and zinc-contaminated regions of the Eagle River than in fish from an uncontaminated control site. Histological analyses revealed that interrenal cells are more stimulated (exhibiting both hypertrophy and hyperplasia) in fish living in contaminated sites than interrenal cells of fish at the control site. These results suggest that the hypothalamo-pituitary-interrenal (HPI) axis of fish living in the metal-contaminated water shows evidence of chronic stimulation. We suggest that assessment of these parameters of the HPI axis may be useful indices of chronic environmental stress in trout. PMID- 9405111 TI - Melatonin and exposure to constant light/darkness affects ovarian follicular kinetics and estrous cycle in Indian desert gerbil Meriones hurrianae. AB - Melatonin mediates photoperiodic influence on reproduction and constant light and darkness affect pineal biosynthesis of melatonin. The present study was undertaken to assess the effects of melatonin and drastic photoperiodic changes on reproduction in a tropical desert species with a fossorial lifestyle. Ovarian follicular kinetics and estrous cycle were studied in the Indian desert gerbil Meriones hurrianae, after treatment with melatonin and exposure to constant light (LL) and darkness (DD) regimes. Melatonin treatment increased (P < 0.001) ovarian weights without changing the uterine weights. While exposure to LL decreased (P < 0.001) both ovarian and uterine weights, exposure to DD had no effect on these weights. Follicular kinetics of growing and regressing follicles revealed that ovaries of melatonin-treated and DD-exposed animals had significantly more growing follicles. Melatonin treatment increased all types of growing follicles, especially antral and Graafian follicles. Exposure to DD increased all types of growing follicles, with the medium sized antral and Graafian follicles being significant (P < 0.01). In contrast to stimulation of follicular growth by melatonin and DD, LL caused regression of all stages of follicular growth and also reduced the number of small preantral follicles. Melatonin treatment increased (P < 0.001) the length of estrous cycle (5.08 to 7.29 days). Gerbils treated with melatonin, exposed to LL and DD, had a longer (P < 0.001) metestrus. Animals held in LL, had the least number (P < 0.001) of estrous smears (1 in 30 days). The results suggest that melatonin is involved in growth of ovarian follicles in the Indian desert gerbil. PMID- 9405112 TI - Rhythmic steroidogenesis by the prothoracic glands of the insect Rhodnius prolixus in the absence of rhythmic neuropeptide input: implications for the role of prothoracicotropic hormone. AB - Circadian rhythms have been reported both in the synthesis of the insect steroid moulting hormones (ecdysteroids) by the prothoracic glands (PGs) and in the release of the cerebral neuropeptide, prothoracicotropic hormone (PTTH). PTTH is known to activate steroidogenesis early in development, but the function of continued rhythmic release is unknown. The functional relationship between these two hormonal rhythms was examined. We report the properties of the rhythm of steroidogenesis by PGs of animals in which PTTH release was prevented by decapitation or by injection of a sublethal dose of tetrodotoxin. Rhythmic steroidogenesis by PGs was maintained in both cases; the rhythm retained entrainment to a light-dark cycle and free-ran in continuous light or darkness. It is inferred that rhythmic neuropeptide input is not required to drive rhythmic steroidogenesis and that in its absence, steroidogenesis becomes entrained by light cues. In both decapitated and paralyzed animals, the rhythm of steroidogenesis showed a reversal of phase from that of intact animals under all conditions of illumination tested. We infer that the rhythm of PTTH appears to entrain rhythmic steroidogenesis and entrainment by PTTH dominates entrainment by light in vivo. Similarities to other circadian systems are discussed, in which neurochemical agents entrain overt rhythms to a phase displaced by 12 hr from that for light. It is concluded that the function of PTTH is not confined to initial activation of steroidogenesis early in development, as previously thought, but continues throughout development as a central element in the circadian organization of the endocrine system that regulates development. PMID- 9405113 TI - Seasonal changes in follicle-stimulating hormone in free-living great tits. AB - We describe the annual cycle in plasma levels of FSH in free-living male and female great tits from southwest Sweden. Both juvenile (here defined as first time breeders or birds <1 year old) and adult great tits, of both sexes, showed clear annual cycles with three periods of elevated FSH levels: period from territorial establishment till end of breeding (mid-March to June), October (a time when the birds break photorefractoriness), and winter (January-February). Significant differences between ages and between sexes occurred only during March and April (period of territorial establishment and gonadal recrudescens). Male FSH levels increased significantly as early as between early February and early March. Levels continued to increase until mid-April when maximal values had been reached in paired males. Territorial, but unpaired, males had significantly lower plasma levels of FSH in mid-April than did territorial and paired males. After this, FSH levels did not change until levels had decreased to basal in early July. On average, females did not show a vernal increase in FSH levels until early/mid April. However, just as in males, female FSH levels were dependent upon whether she was paired or unpaired. Females having a mate in mid-April had significantly higher FSH levels than did single females. Contrary to the males, females showed a transitory FSH peak during egg-laying. Females showed no differences between other breeding stages, until basal levels were reached during incubation of the second clutch in July. Adult birds (of both sexes) had significantly higher plasma levels of FSH than juvenile birds of the same sex during the period March-April. Furthermore, during this period adult males continually had higher FSH levels than did adult females, and juvenile males continually had higher levels than did juvenile females. In all groups a second period of elevated FSH levels occurred during early October, a time when the great tits break photorefractoriness. All groups showed basal levels during November and December. By January all four groups had increased their circulating levels of FSH to moderately high. This elevated level was maintained during February and was at the same level as that observed in October. PMID- 9405114 TI - Identification and characterization of growth hormone receptors in snakehead fish (Ophiocephalus argus cantor) liver. AB - The specific binding of 125I-labeled fish growth hormone (GH) to hepatic membranes prepared from several freshwater fish was assessed. A high level of growth hormone receptor (GHR) was detected on the hepatic membranes of the snakehead fish (Ophiocephalus argus Cantor). Scatchard analysis of the binding data showed a single class of high affinity binding site with a binding affinity (Ka) of 1.45 +/- 0.23 x 10(9) M-1 and a binding capacity (Bmax) of 198 +/- 57 fmol/mg protein. The binding was specific for fish GH and was saturable. In addition, the specific binding was temperature- and time-dependent, reaching a steady state after 16 hr of incubation at 25 degrees . The molecular weight of GHR as measured by Sephadex G-200 column chromatography and Western blot analysis using a monoclonal antibody (Mab263) against GHR was found to be 200-400 and 90 93 kDa, respectively. Two bands at 65 and 89 kDa were identified in ligand crosslinking studies of membrane receptors. A sensitive teleost GH radioreceptor assay (RRA) was developed, using recombinant fish GH and a membrane preparation from snakehead fish liver, capable of measuring bioactive GH in fish sera or other samples. PMID- 9405115 TI - Melatonin does not prevent long photoperiod stimulation of secondary sexual characters in the male three-spined stickleback Gasterosteus aculeatus. AB - Breeding in the three-spined stickleback is stimulated by long but not by short photoperiods in many seasons. The aim of the present study was to test the hypothesis that melatonin plays a role in the inhibitory effect of short photoperiod in this species. Adult nonbreeding males were kept either under constant light (Experiment 1) or under a stimulatory long photoperiod (16L 8D, Experiment 2), in water containing 0, 20, or 80 microg/liter melatonin for 16 hr/day for 28 days during the spring. These melatonin treatments were intended to simulate the daily melatonin pattern of a nonstimulatory short photoperiod. In the second experiment, fish were also kept under a nonstimulatory short photoperiod (8L 16D). In the natural breeding season the only germ cells found in the stickleback testes are spermatozoa and spermatogonia, a condition found in many fish under all treatments. In the first experiment, spermatogenesis was not influenced by melatonin. However, testes also containing spermatocytes and spermatids were more common in fish kept under 8L 16D and fish treated with 80 microg/liter melatonin than in 16L 8D controls in Experiment 2. Kidney hypertrophy, an androgen-dependent male secondary sexual characteristic in the stickleback, appeared in most males kept under constant light or 16L 8D and was not influenced by melatonin treatment. In contrast, control males kept under 8L 16D in Experiment 2 did not display kidney hypertrophy. Therefore, the presence of an extended period of elevated melatonin did not prevent the stimulatory effects of long photoperiod on development of this secondary sexual characteristic in the stickleback. PMID- 9405116 TI - Lipopolysaccharide-induced hyperglycemia is mediated by CHH release in crustaceans. AB - Septicemia in crustaceans may occur occasionally due to Gram-negative opportunistic bacteria, especially under conditions of intensive aquaculture. The lipopolysaccharide (LPS) endotoxin induces in mammals septic shock and the activation by LPS of hormone release through the hypothalamo-pituitary axis is well known. In crustaceans an increase in circulating Crustacean hyperglycemic hormone and hyperglycemia are reported to result from exposure to several environmental stressors but the metabolic and hormonal effects of LPS in vivo are undescribed. A sublethal dose of LPS (Sigma, Escherichia coli 0111:B4) was injected into at least five individuals of species representative of crustacean taxa and life habits: Squilla mantis (Stomatopoda); the Decapoda Crangon crangon and Palaemon elegans (Caridea), Nephrops norvegicus (Astacidea), Munida rugosa and Paguristes oculatus (Anomura), Pilumnus hirtellus, Macropipus vernalis, Parthenope massena, and Ilia nucleus (Brachyura). Within 3 hr an increase in blood sugar developed ranging from 26.00 +/- 8.37 sd mg/dl in M. rugosa to 201.50 +/- 95. 91 sd mg/dl in P. oculatus and a significant increase of 79% in M. rugosa up to 1300% in P. hirtellus over control levels was observed. The involvement of eyestalk hormones in this generalized response was tested on S. mantis, M. vernalis, and P. elegans; LPS injected into eyestalkless animals did not elicit a significant hyperglycemic response compared with saline-injected controls. Eyestalkless animals injected with one eyestalk equivalent homogenate in saline from untreated animals did show a change in color from red to normal likely due to red pigment concentrating hormone and a hyperglycemic response within 2 hr. Eyestalkless animals injected with homogenate from LPS-treated shrimps showed the change in color but not the hyperglycemic response. It is concluded that LPS directly, or cytokines circulated upon challenge by the endotoxin, may act on the medulla terminalis X-organ-sinus gland complex and release CHH selectively eliciting an hyperglycemic stress response, after which CHH stores become relatively depleted. PMID- 9405117 TI - Isolation and characterization of the female-specific protein (vitellogenin) in mature female hemolymph of the freshwater prawn, Macrobrachium rosenbergii: comparison with ovarian vitellin. AB - Purification and characterization of the female-specific protein (vitellogenin) from the hemolymph of mature female prawn, Macrobrachium rosenbergii, were the objectives of this study. The comparison of biochemical characteristics between vitellogenin and ovarian vitellin was also conducted. Hemolymph vitellogenin was purified with DEAE, hydroxylapatite, and another DEAE chromatographic column. The specific protein (vitellogenin) was shown in the fractions of chromatographic columns on the basis of ELISA, Western blotting, and immunoprecipitation. A purified vitellogenin was obtained with an apparent molecular weight of 700 kDa as determined by PAGE. The purified vitellogenin was considered as a lipoglycoprotein on the basis of staining data. Three subunits (170, 100, and 89 kDa) in purified vitellogenin and two subunits (100 and 89 kDa) in vitellin were detected with SDS-PAGE. Nondisulfide bonds were found in the binding of polypeptide subunits. Only the 89-kDa subunit was a glycopolypeptide in both vitellogenin and vitellin. The amino acid composition of vitellogenin differed from that of vitellin in a few amino acids. Eight amino acid sequences from the N terminal end of 89- and 100-kDa subunits were determined and they were identical between vitellogenin and vitellin. Seven amino acid sequence from the N-terminal end of the 170-kDa subunit were also identical to the 100-kDa subunit. Purified vitellogenin was more susceptible to precipitation in a solution with low ionic strength than vitellin. This study suggests a close relationship between vitellogenin and vitellin in M. rosenbergii in their biochemical characteristics. PMID- 9405118 TI - Stage-dependent changes in adrenal steroids and catecholamines during development in Xenopus laevis. AB - Changes in adrenal hormones during the complete developmental cycle from egg to juvenile were investigated in the amphibian Xenopus laevis. Whole-body concentrations of the adrenal steroids corticosterone (B), and aldosterone (Aldo) were determined by radioimmunoassay and those of the adrenal catecholamines epinephrine (E), norepinephrine (NE), and dopamine (D) were determined by HPLC. In addition, the catecholamine-synthesizing enzymes tyrosine hydroxylase, dopamine beta-hydroxylase, and phenylethanolamine N-methyltransferase were immunocytochemically localized for the characterization of chromaffin adrenal cells. B and Aldo were not detectable in the whole body before hatching. B levels rose earlier than Aldo levels from stage 36 onward. B had already peaked at stage 46, whereas the largest amounts of Aldo were found at stage 54. After peaking, both steroids decreased gradually to 2.7 +/- 0.62 (B) and 0.4 +/- 0.1 (Aldo) ng/g body wt (mean +/- SEM, n = 10) in juvenile animals. E, NE, and D were detected just after hatching, when E and D showed an early peak at stage 40. E and NE increased moderately during development and demonstrated a sharp increase at the end of metamorphosis from stages 62 onward to 14.4 +/- 1.7 (E) and 34.1 +/- 4.67 (NE) ng/g body wt (mean +/- SEM, n = 6). Interestingly, D levels had a distinct pattern, because concentrations of D remained lower than those of NE and E over nearly the complete development, but showed a dramatic rise during the latest stages, reaching 707 +/- 54 ng/g body wt in juveniles. This dramatic shift in catecholamine levels was confirmed by immunocytochemistry in parallel. A large increase in chromaffin cells labeled with tyrosine hydroxylase immunoreactivity occurred in the latest developmental stages. The catabolic rates for all catecholamines in vivo were similar, which indicates that the different levels are due to various rates of synthesis. Thus, adrenal corticosteroids as well as catecholamines may have regulatory effects during premetamorphosis and metamorphic climax. PMID- 9405119 TI - Corticosterone secretion in response to capture and handling in free-living red eared slider turtles. AB - The corticosterone response to capture and handling was measured in free-living red-eared slider turtles, Trachemys scripta elegans. To determine the ability of this species to exhibit this endocrine response, slider turtles were bled at the time of removal from hoop nets and again at 30 and 60 min following capture to create plasma profiles of acute corticosterone secretion from individuals. Plasma corticosterone concentration increased significantly with handling time. The greatest rise in corticosterone was within the first 30 min following capture and handling, with this rate of increase declining over the next 30 min of restraint. There was no correlation between corticosterone levels at the time of capture and the length of time it took to get the sample if the sample was taken within the first 10 min after capture. However, when these samples were included with those taken from other turtles sampled 11 to 25 min after capture, hormone levels were significantly correlated with handling time. This suggests that the critical time to obtain an initial sample that best represents the predisturbance level in slider turtles is within 10 min. There was no correlation between the turtles' energetic condition and initial corticosterone concentrations. Plasma corticosterone values at all sampling times were comparable to those observed in other reptile species. The results from this study may be used to investigate the effects of unpredictable resources on reproductive success and survival in freshwater turtles. PMID- 9405120 TI - Induction of male-type gonadotropin secretion by implantation of 11 ketotestosterone in female goldfish. AB - In goldfish, plasma gonadotropin levels increase during spawning in both males and females (GTH surge). A female-typical GTH surge induces ovulation (ovulatory surge), and a male-typical surge triggers milt production in response to sex pheromones released from ovulatory females. This study examined whether the male typical GTH surge occurs in adult females that are implanted with 11 ketotestosterone (KT), which induces male-typical sexual behavior in adult female goldfish. When KT-implanted females were exposed to ovulatory females, a GTH surge occurred without ovulation. No GTH surge was observed when KT-females were exposed to nonovulatory females. The GTH secretion in KT-females was further characterized by exposure to 17alpha,20beta-dihydroxy-4-pregnen-3-one (17,20-P), a female sex pheromone that induces the GTH surge in males. Exposure to waterborne 17,20-P caused an elevation of GTH levels in KT-females as well as in males. The elevation of GTH levels induced by 17,20-P exposure was abolished when the KT-females were rendered anosmic. Unlike the female-typical ovulatory GTH surge that occurs in synchrony with photoperiod and peaks in the dark phase of the day, the 17,20-P-induced surge did not show a peak in the dark phase. These results indicate that the GTH surge in KT-females is a male-typical surge. Together with a previous study showing KT-induced behavioral masculinization (N. E. Stacey and M. Kobayashi, 1996, Horm. Behav. 30, 434-445), this adult gonochoristic species was shown to possess sexual plasticity of brain function in behavior and GTH secretion in response to sex steroid. PMID- 9405121 TI - Somatostatin inhibition of growth hormone release in goldfish: possible targets of intracellular mechanisms of action. AB - Previous studies have demonstrated that growth hormone (GH) release in goldfish is under the stimulatory control of gonadotropin-releasing hormone (GnRH) and dopamine and the inhibitory control of somatostatin (SRIF). GnRH stimulation is mediated through protein kinase C (PKC)- and calcium-dependent mechanisms, whereas dopamine D1 receptor activation increases GH secretion through cyclic (c) AMP-dependent intracellular signal transduction pathways. In this study, the mechanisms of SRIF inhibition on GH secretion were examined using primary cultures of dispersed goldfish pituitary cells in static incubation. Application of 1 microM SRIF inhibited the GH-release responses to 100 nM salmon GnRH, 100 nM chicken GnRH-II, and 1 microM SKF38393, a D1 agonist. These results indicate that inhibitory action of SRIF on stimulated GH release is direct, at the level of the pituitary cells. Addition of SRIF reduced the GH release responses to two activators of PKC (100 microM dioctanoyl glycerol and 100 nM tetradecanoyl phorbol acetate) and to two ionophores (10 microM A23187 and 10 microM ionomycin). Similarly, SRIF abolished the GH responses to an activator of adenylate cyclase (10 microM forskolin), a membrane-permeant cAMP analog (1 mM 8 bromo-cAMP), and a voltage-sensitive calcium channel agonist (1 microM Bay K 8644). Taken together, these observations indicate that the inhibitory actions of SRIF on D1- and GnRH-stimulated GH release can be exerted at sites distal to cAMP production and PKC activation, respectively. SRIF also exerts its effect at sites distal to calcium mobilization. Since SRIF inhibition was more effective against Bay K 8644-induced response than against ionophore-induced GH response, an inhibitory action at the level of extracellular calcium entry through voltage sensitive channels is also possible. PMID- 9405122 TI - Salmon thyroid-stimulating hormone: isolation, characterization, and development of a radioimmunoassay. AB - Coho salmon (Oncorhynchus kisutch) thyroid-stimulating hormone (TSH) was isolated by ethanol extraction of pituitary glands from mature coho salmon. Extraction was followed by gel-filtration chromatography on Sephadex G-100 superfine, anion exchange chromatography on a Whatman DE-52 column, and finally by reverse-phase high-performance liquid chromatography. Fractions were monitored for TSH content by a homologous in vivo bioassay and by immunoblots using anti-human TSH beta subunit antisera. In vivo treatment of coho salmon parr with coho salmon TSH caused a dose-dependent increase in plasma thyroxine level similar to that induced by bovine TSH. The N-terminal sequence (25 residues) of the salmon TSH beta subunit has 56% sequence identity to that of human TSH beta subunit and is identical to the deduced amino acid sequence of trout TSH beta subunit. The N terminal sequence (25 residues) of the salmon TSH alpha subunit is identical to gonadotropin alpha-II subunit. Molecular sizes of the alpha and beta subunits are 18,000 and 24,000 daltons, respectively, as estimated by SDS-PAGE. Antiserum generated against salmon TSH, which was preadsorbed with alpha subunit using an alpha-subunit affinity column, detected only salmon TSH beta subunit by immunoblot and specifically stained thyrotropin-producing cells of the pituitary gland. A homologous radioimmunoassay (RIA) was developed using purified salmon TSH standard, iodinated TSH beta subunit, and antiserum generated against salmon TSH. Cross-reactivities of GTH I, GTH II, GTH I beta and GTH II beta subunits, alpha subunit, growth hormone, prolactin, and somatolactin were less than 1%. Displacement curves for serial dilutions of plasma and pituitary extracts of various salmonid species, as well as coho salmon pituitary cell culture medium, were parallel to the coho salmon TSH standards. In contrast, plasma of hypophysectomized juvenile coho salmon and pituitary extracts of Pacific tomcod (Microgadus proximus) did not displace bound radiolabeled salmon TSH. Finally, in vivo injection of juvenile coho salmon with triiodothyronine decreased plasma TSH levels, whereas the goitrogen, methimazole, increased plasma TSH levels. Injection of gonadotropin-releasing hormone agonist did not alter plasma TSH. These data suggest that the RIA is specific for TSH and confirm a negative feedback relationship between the thyroid hormones and TSH. PMID- 9405123 TI - Interrenal stress responsiveness of tilapia (Oreochromis mossambicus) is impaired by dietary exposure to PCB 126. AB - Activation of the hypothalamus-pituitary-interrenal (HPI) axis is characteristic of stress responses, which may result from a variety of environmental challenges. To investigate whether the stress response, and in particular the HPI axis, in tilapia (Oreochromis mossambicus) is compromised by short-term exposure to PCB 126, fish of both sexes were fed diets containing PCB 126 (50 microg/kg fish . day) for 5 days. In the first approach, which was performed twice, fish were acutely stressed for periods varying between 1 and 30 min at the end of the exposure period; in the second approach fish were sampled at the end of the exposure period either at rest or after 2 h of stress (confinement). After 5 days, the body weights in all experiments were significantly lower in PCB-fed fish than in control fish. There were no changes in basal plasma glucose levels, plasma ion concentrations, or branchial, renal, and intestinal Na,K-ATPase activity following PCB exposure. In the first experimental approach, in which fish experienced acute sampling stress, plasma cortisol levels reached lower levels in PCB-fed fish than in controls. This suggests an impaired ability to acutely activate interrenal steroidogenesis in PCB-treated tilapia. Adrenocorticotropic hormone (ACTH)- and cAMP-stimulated in vitro cortisol release from superfused head kidneys was lower in tissues from tilapia exposed to PCB 126 than in tissues from control animals. This effect persisted after 24 h in vitro, which, together with the high PCB 126 concentrations measured in the head kidneys of PCB-fed fish, may indicate direct toxic effects on the interrenal cells. The second experimental approach demonstrated that basal plasma cortisol and ACTH levels were not influenced by PCB treatment, but that the basal ACTH content of the rostral pars distalis (RPD) of the pituitary gland of PCB-fed fish was lower than that of control fish. After 2 h confinement, plasma cortisol levels and ACTH content of the RPD rose to similar values in both groups, whereas plasma ACTH levels were higher in confined PCB-fed fish than in confined controls. PCB-fed fish showed a lower hyperglycemic response to confinement than control fish. Confinement resulted in similarly elevated renal and intestinal Na,K-ATPase activities in both PCB-fed and control fish; branchial enzyme activities were not affected. Since PCB did not affect Na,K-ATPase activities and plasma ion concentrations, it is concluded that the effects of PCB 126 on the HPI axis in tilapia are not secondary to ionoregulatory dysfunction. PMID- 9405124 TI - Adrenocortical and adrenomedullary homologs in eight species of adult and developing teleosts: morphology, histology, and immunohistochemistry. AB - Morphology, histology, and immunohistochemistry of the adrenocortical and adrenomedullary homologs (adrenal glands) of the following developing and adult teleosts were examined: Salmoniformes-Oncorhynchus mykiss (rainbow trout), Salmo trutta fario (brown trout), Coregonus lavaretus (white fish); Cyprinodontiformes Gambusia affinis (mosquito fish). Perciformes-Dicentrarchus labrax (sea bass), Sparus aurata (sea bream), Diplodus sargus (white bream), Oblada melanura (saddled bream). The anatomical relationships of the gland with the renal system and venous vessels were also noted. In adults of all species steroidogenic and catecholaminergic chromaffin cells were found in the head kidney, which is pronephric in origin and subsequently transformed into a hematopoietic lymphatic organ. In Perciformes, chromaffin cells are distributed around the anterior and posterior cardinal veins and ducts of Cuvier; in Salmoniformes, around the posterior cardinal veins and in the hematopoietic tissue; and in G. affinis, around the ducts of Cuvier and posterior cardinal veins, while a few are visible also around the sinus venosus. In Perciformes and Salmoniformes, numerous chromaffin cells are also present in the posterior kidney, derived from the opisthonephros, in contact with the caudal vein. Steroidogenic cells are always confined to the head kidney. During development chromaffin and steroidogenic cells appear early after hatching in the pronephric kidney, at the level of the ducts of Cuvier and of the cephalic part of the posterior cardinal veins. Later, chromaffin cells in Perciformes reach the anterior cardinal veins, and subsequently, in both Perciformes and Salmoniformes, they reach the developing posterior kidney. Their localization along the posterior kidney is still in progress about 4 months after hatching and is completed about a year after hatching. These findings support the concept that the structure of the adrenal gland in teleosts is intermediate between that of the other actinopterygians and that of tetrapods. The development differs from that of tetrapods in that it occurs mainly in the pronephros and only later do chromaffin cells reach the opisthonephric kidney. PMID- 9405129 TI - Transformation of sensory signals into commands for saccadic eye movements: a neural network study. AB - A biological plausible neural network which simulated the input-output transformation performed by primates during saccadic eye movements is constructed using a selective attention module and multi-layered neural networks with improved back propagation and a competitive learning algorithm. Simulation results show that the trained model can make fine saccades directed by the target. Representations and processing mechanisms in the saccade system are investigated. The features of most hidden units resemble those that have been observed in physiological recordings of neurons in primates visual cortex. The hidden layer even developed structures similar to those of area 7a. PMID- 9405130 TI - Hydroxyaluminosilicates and acute aluminium toxicity in fish AB - The essentiality of silicon in biology might be explained in the terms of its chemistry with aluminium. In a previous study we demonstrated the elimination of acute aluminium toxicity in fish by silicon. We suggested that the reaction of silicic acid with aluminium to form hydroxyaluminosilicates reduced the biological availability, and hence toxicity, of aluminium. Though assumed in a burgeoning number of studies and contended in others this detoxification mechanism has remained unproven. Herein we have tested the toxicity of hydroxyaluminosilicates in fish and in doing so we have provided evidence which strongly supports a role for hydroxyaluminiosilicates in the elimination of acute aluminium toxicity in fish by silicon.Copyright 1997 Academic Press Limited Copyright 1997 Academic Press Limited PMID- 9405131 TI - Deriving shape space parameters from immunological data. AB - We present a method for deriving shape space parameters that are consistent with immunological data, and illustrate the method by deriving shape space parameters for a model of cross-reactive memory. Cross-reactive memory responses occur when the immune system is primed by one strain of a pathogen and challenged with a related, but different, strain. Much of the nature of a cross-reactive response is determined by the quantity and distribution of the memory cells, raised to the primary antigen, that cross-react with the secondary antigen. B cells with above threshold affinity for an antigen lie in a region of shape space that we call a ball of stimulation. In a cross-reactive response, the intersection of the balls of stimulation of the primary and secondary antigens contains the cross-reactive B cells and thus determines the degree of cross-reactivity between the antigens. We derive formulas for the volume of intersection of balls of stimulation in different shape spaces and show that the parameters of shape space, such as its dimensionality, have a large impact on the number of B cells in the intersection. The application of our method for driving shape space parameters indicates that, for Hamming shape spaces, 20 to 25 dimensions, a three or four letter alphabet, and balls of stimulation of radius five or six, are choices that match the experimental data. For Euclidean shape spaces, five to eight dimensions and balls of stimulation with radius about 20% of the radius of the whole space, match the experimental data. PMID- 9405132 TI - Direction of protein biosynthesis as a reflection of the prebiotic environment AB - Systematical analyses of the folding pathways of simple polypeptides show that for systems containing L-stereoisomers their natural tendency to be right-handed abakes when the polypeptide terminal are naturally charged. This disadvantageous effect is minimized by an introduction of charged amino acid residues into the polypeptide chain at the beginning of its synthesis. Since in the rebiotic environment negatively charged amino acids were more prominent than positively charged ones, we conclude that the fixed direction of protein biosynthesis might be a result of "a broken symmetry" of amino acid charges.Copyright 1997 Academic Press Limited Copyright 1997 Academic Press Limited PMID- 9405133 TI - A general model of pheromone transmission within honey bee hives AB - Many activities in a honey bee hive are thought to be regulated by non-volatile pheromones produced by the queen. These pheromones are transmitted among the workers both by direct contact and by deposition on the retrieval from the wax substrate of the hive. This paper presents a general model for the movement of these pheromones through a honey bee colony.The specific pheromone used for the model is produced by honey bee queens, and is thought to play a determining role in colony swarming. Thus, the results presented here are of interest to both beekeepers, for whom swarming represents the loss of a valuable resource, and bee researchers, who are interested in swarming as an example of social coordination.The model consists of a system of stochastic differential equations describing the evolution of the pheromone level and position of each bee in the hive. The equations are studied using a numerical simulation. The results of the simulations show that hive crowding can have a significant effect on pheromone transmission, and that this effect is not simply a dilution of similar quantities of pheromone among more workers. Thus, restricting worker movement induced by crowding may have a strong effect on pheromone transmission and, therefore, advance queen rearing and swarming. Copyright 1997 Academic Press Limited Copyright 1997 Academic Press Limited PMID- 9405134 TI - About a symmetry of the genetic code. AB - Considering the three codonic positions as independent, it is possible to explain the grouping of the 64 trinucleotides (without AAA, TTT, CCC and GGG) into three equal sets T0, T1 and T2 according to their preferable reading frame (0, 1 or 2) in coding sequences. Supposing that the two complementarity strands of DNA are coding, it is demonstrated that the complementary of a codon is classified in to the same set, which has been observed statistically (with a few exceptions) in the coding sequences by Arques & Michel [(1996) J. theor. Biol. 182, 45-58] and Arques et al. [(1997) J. theor. Biol. 185, 241-253]. Finally, the circular property of the code pointed out by these authors is demonstrated, and a direct consequence to biological considerations of these properties is discussed. PMID- 9405135 TI - The n-player war of attrition and territorial groups. AB - The choices made by juveniles, in territorial species, between dispersing and remaining in the natal territory, can be modelled as a simple multi-player evolutionary game, related to the well-known War of Attrition [Maynard Smith, J. (1974) J. theor. Biol. 47, 209-221; Haigh J. & Cannings, C. (1989) Acta Applicandae Mathematicae 14, 59-74]. The game is shown to have a unique evolutionarily stable strategy, involving a random choice between dispersing early in the game and staying indefinitely. An example is given, involving badgers (Meles meles), in which the key factor affecting the pay-off in the game is the possibility of inheriting the territory on the death of the current holders. The example indicates the sensitivity of the size of the group occupying the territory to the mortality rate among dispersers. PMID- 9405136 TI - Deciphering the language of the genome. AB - The non-coding DNA in eukaryotic genomes encodes a language which programs organismal growth and development. We show that a linguistic and cryptographic approach can be used to deduce the syntax of this programming language for gene regulation and to compile a dictionary of enhancers which form its words. PMID- 9405137 TI - Mechanistic modeling of prokaryotic mRNA decay. AB - A mechanistic model of gene expression was developed to test three prevailing and sliding prokaryotic mRNA decay theories: ribosome protection of mRNA from endonucleases, 5' binding and sliding of endonucleases on mRNA, and hybrid 5' binding/ribosome protection. The discrete event simulation incorporates the molecular events that determine both cellular mRNA and protein levels. A Monte Carlo technique was used to approximate the inherent randomness of the molecular processes involved in gene expression. Each of the decay theories was tested for the ability to predict the effects of ribosome loading and translation rate on mRNA stability as well as the observed 5' to 3' directionality of mRNA decay. The modeling results show that the hybrid decay mechanism best predicts the experimentally-observed mRNA decay behaviors. The 5' binding mechanism fails to adequately predict the sensitivity of mRNA stability to changes in translation rate and ribosome loading, while the ribosome protection mechanism does not correctly predict 5' to 3' decay directionality. In addition to discriminating between the three decay theories, the simulations provide insights into hybrid decay mechanism specific details such as RNase binding and cleavage characteristics. Finally, we discuss the application of the current mechanistic model for analysing and predicting expression from more complex genetic systems. PMID- 9405138 TI - A general technique for computing evolutionarily stable strategies based on errors in decision-making. AB - Realistic models of contests between animals will often involve a series of state dependent decisions by the contestants. Computation of evolutionarily stable strategies for such state-dependent dynamic games are usually based on damped iterations of the best response map. Typically this map is discontinuous so that iterations may not converge and even if they do converge it may not be clear if the limiting strategy is a Nash equilibrium. We present a general computational technique based on errors in decision making that removes these computational difficulties. We show that the computational technique works for a simple example (the Hawk-Dove game) where an analytic solution is known, and prove general results about the technique for more complex games. It is also argued that there is biological justification for inclusion of the types of errors we have introduced. PMID- 9405139 TI - Three-dimensional structures of proteins involved in programmed cell death. AB - Programmed cell death (apoptosis) is a controlled process by which unwanted cells are selectively eliminated. Several families of proteins including the Bcl-2, tumor necrosis factor receptor 1, and caspase families play essential roles in the regulation, signaling, and execution of the genetic cell death program. The recently described three-dimensional structures of members of these families elucidate the structural basis of their functions and provide insights into the mechanisms by which these proteins regulate apoptosis. PMID- 9405140 TI - DNA-binding specificity of the homeodomain-leucine zipper domain. AB - Homeodomain-leucine zipper (HD-Zip) proteins are putative transcription factors identified only in plants. The study of the DNA-binding properties of the ATHB-1 and -2 HD-Zip (HD-Zip-1 and -2) domains showed that they interact with DNA as homodimers and recognize two distinct 9 bp pseudopalindromic sequences, CAAT(A/T)ATTG (BS-1) and CAAT(G/C)ATTG (BS-2), respectively, as determined by selecting high-affinity binding sites from random-sequence DNA. Here, we report a mutational analysis of the HD-Zip-2 domain. We determined that conserved amino acid residues of helix 3, Val47 and Asn51, and Arg55 are essential for the DNA binding activity of the HD-Zip-2 domain. We demonstrated that the preferential recognition of a G/C base-pair at the central position by the HD-Zip-2 domain is abolished either by the replacement of Arg55 with lysine or by the substitution of Glu46 and Thr56 with the corresponding residues of the HD-Zip-1 domain (alanine and tryptophan, respectively). In contrast, substitution of Arg55 with lysine in the HD-Zip-1 domain significantly reduced DNA-binding activity without changing the specificity of recognition. Finally, we determined that differences in residues outside helix 3 further contribute to the DNA-binding specificity of the HD-Zip domain. Taken together, the data strongly suggest that the preferential recognition of BS-2 and -1 by the HD-Zip-2 and -1 domains, respectively, may be attributable to a distinct orientation of the side-chain of Arg55 in these two domains. PMID- 9405141 TI - Visualization of caldesmon on smooth muscle thin filaments. AB - Caldesmon, a narrow, elongated actin-binding protein, is found in both nonmuscle and smooth muscle cells. It inhibits actomyosin ATPase and filament severing in vitro, and is thus a putative regulatory protein. To elucidate its function, we have used electron microscopy and three-dimensional image reconstruction to reveal the location of caldesmon on isolated smooth muscle thin filaments. Caldesmon density was clearly delineated in reconstructions and found to occur peripherally, on the extreme outer edge of actin subdomains-1 and 2, without making obvious contacts with tropomyosin strands on the inner domains of actin. When the reconstructions were fitted to the atomic model of F-actin, caldesmon appeared to cover potentially weak sites of myosin interaction with actin, while, together with tropomyosin, it flanked strong sites of myosin interaction, without covering them. These interactions are unlike those of troponin-tropomyosin and therefore inhibition of actomyosin ATPase by caldesmon-tropomyosin and by troponin-tropomyosin cannot occur in the same way. The location of caldesmon would allow it to compete with a number of cellular actin-binding proteins, including those known to sever or sequester actin. PMID- 9405142 TI - The universal stress protein, UspA, of Escherichia coli is phosphorylated in response to stasis. AB - Transcriptional induction of the uspA gene of Escherichia coli occurs whenever conditions cause growth arrest and cells deficient in UspA survive poorly in stationary phase. We demonstrate that the product of uspA is a serine and threonine phosphoprotein. In vivo, three isoforms of UspA were detected, two of which were phosphorylated as determined by alkaline phosphatase treatment; in vitro, phosphorylation with [gamma-32P]ATP yielded two radioactive UspA isoforms. The phosphorylated isoforms were barely visible in growing cells but one increased during starvation conditions causing growth arrest. This phosphorylation is dependent on the o591 gene, which encodes an autophosphorylating tyrosine phosphoprotein and which is involved in the synthesis or modification of six other proteins. In vitro, UspA undergoes a rapid and dynamic autophosphorylation, as shown by chase experiments with GTP or ATP as phosphate donors. PMID- 9405143 TI - Identification of functional surfaces of the zinc binding domains of intracellular receptors. AB - Transcriptional regulatory factor complexes assemble on genomic response elements to control gene expression. To gain insights on the surfaces that determine this assembly in the zinc binding domains from intracellular receptors, we systematically analyzed the variations in sequence and function of those domains in the context of their invariant fold. Taking the intracellular receptor superfamily as a whole revealed a hierarchy of amino acid residues along the DNA interface that correlated with response element binding specificity. When only steroid receptors were considered, two additional sites appeared: the known dimer interface, and a novel putative interface suitably located to contact regulatory factors bound to the free face of palindromic response elements commonly used by steroid receptors. Surprisingly, retinoic acid receptors, not known to bind palindromic response elements, contain both of these surfaces, implying that they may dimerize at palindromic elements under some circumstances. This work extends Evolutionary Trace analysis of functional surfaces to protein-DNA interactions, suggests how coordinated exchange of trace residues may predictably switch binding specificity, and demonstrates how to detect functional surfaces that are not apparent from sequence comparison alone. PMID- 9405144 TI - A novel highly reproducible quantitative competitve RT PCR system. AB - Small changes in plasma fibrinogen concentration are often clinically important. To detect changes in fibrinogen mRNA level, we have designed a novel quantitative competitive reverse transcriptase (RT) PCR system, based on our recent understanding that an important factor causing imprecision with RT PCR is a change in the initial ratio of target to standard RNA templates during reverse transcription. The system comprises: (1) titration of a fixed amount of standard RNA with four different amounts of total RNA in each tube during cDNA synthesis; (2) amplification of each of the four cDNAs with four consecutive PCR cycles; and (3) quality control reactions in which synthesised quality control RNA (rather than total RNA) is added to the standard RNA under the same RT PCR conditions as the assay reaction. To obtain reproducible data, we suggest three criteria for analysis of RT PCR results. Employing these criteria, reproducibility of RT PCR was markedly improved with an inter-assay CV of 6.5% (n=5). Using this system, we are able to detect reduction in mRNA levels of all fibrinogen genes caused by dietary protein restriction in rats after weaning (alpha 39%, p= 0. 028; beta 55%, p = 0.017; gamma 35 %, p= 0.038), with a parallel reduction of plasma fibrinogen concentration (mean (+/-SD), 157(+/-19) versus 130(+/-14) mg/dl, p=0.006). PMID- 9405145 TI - A group II self-splicing intron from the brown alga Pylaiella littoralis is active at unusually low magnesium concentrations and forms populations of molecules with a uniform conformation. AB - We have investigated the reactivity of three of the seven group II introns encoded by the mitochondrial genome of the brown alga Pylaiella littoralis. While the first intron in the protein-coding cox1 gene could not be induced to self splice under any of the conditions tested, the first two introns in the gene encoding the large ribosomal subunit are reactive in vitro and splice primarily by the standard group II two-step transesterification pathway. Intron 2 proved to be of exceptional interest, because in contrast to all group II molecules known so far, its optimal magnesium concentration is less than 10 mM and it still carries out accurate splicing at concentrations as low as 0.1 mM magnesium. Analysis of reaction products under optimal conditions showed no evidence of hydrolysis at the 5' splice site and up to 90% of precursor molecules could be converted into excised lariat intron, which migrated as a single band on non denaturing polyacrylamide gels. Absorbance versus temperature profiles generated from the lariat form of intron 2 reveal the existence of an early melting component, the amplitude of which does not depend on the way the molecules were purified, i.e. with or without a denaturation step. This highly cooperative transition, whose position along the temperature axis changes with the concentration of magnesium, is proposed to consist of the unfolding of the tertiary structure of the molecule. We conclude that group II introns, which are the largest known ribozymes, can form conformationally homogeneous populations of molecules suitable for physical-chemical studies of higher-order structure. PMID- 9405146 TI - Complexes at the replication origin of Bacillus subtilis with homologous and heterologous DnaA protein. AB - The initial steps in the formation of the initiation complex at oriC of Bacillus subtilis were analyzed with special emphasis on the exchangeability of B. subtilis DnaA protein by DnaA of Escherichia coli. The DNA binding domain of B. subtilis DnaA protein was localized in the 93 C-terminal amino acids. Formation of the "initial complex", as analyzed by electron microscopy, was indistinguishable with B. subtilis DnaA protein or with E. coli DnaA. Similarly, both proteins were able to form loops by interaction of DnaA proteins bound to the DnaA box regions upstream and downstream of the dnaA gene in B. subtilis oriC. The region of local unwinding in the "open complex" was precisely defined. It is located at one side of a region of helical instability, a DNA unwinding element (DUE). Unwinding in oriC could only be catalyzed by the homologous DnaA protein. PMID- 9405147 TI - The 118-135 peptide of the human prion protein forms amyloid fibrils and induces liposome fusion. AB - The prion protein (PrPC) is a glycoprotein of unknown function normally found at the surface of neurons and of glial cells. It is involved in diseases such as bovine spongiform encephalopathy, and Creutzfeldt-Jakob disease in the human, where PrPC is converted into an altered form (termed PrPSc). PrPSc is highly resistant towards proteolytic degradation and accumulates in the central nervous system of affected individuals. By analogy with the pathological events occuring during the development of Alzheimer's disease, controverses still exist regarding the relationship between amyloidogenesis, prion aggregation and neuronal loss. To unravel the mechanism of PrP neurotoxicity and understand the interaction of PrP with cellular membranes, a series of natural and variant peptides spanning residues 118 to 135 of PrP was synthesized. The potential of these peptides to induce fusion of unilamellar lipid vesicles was investigated. According to computer modeling calculations, the 120 to 133 domain of PrP is predicted to be a tilted lipid-associating peptide, and to insert in a oblique way into a lipid bilayer through its N-terminal end. In addition to amyloidogenic properties exhibited in vitro by these peptides, peptide-induced vesicle fusion was demonstrated by several techniques, including lipid- and core-mixing assays. Elongation of the 120 to 133 peptide towards the N- and C-terminal ends of the PrP sequence showed that the 118 to 135 PrP peptide has maximal fusogenic properties, while the variant peptides had no effect. Due to their high hydrophobicity, all peptides tested were able to interact with liposomes to induce leakage of encapsulated calcein. We demonstrate also that the propensity of the peptides to fold as an alpha-helix increases their fusogenic activity, thus accounting for the maximal fusogenic activity of the most stable helix at residues 118 to 135. These data suggest that, by analogy with the C-terminal domain of the beta-amyloid peptide, the fusogenic properties exhibited by the prion peptides might contribute to the neurotoxicity of these peptides by destabilizing cellular membranes. PMID- 9405148 TI - X-ray crystal structure and solution fluorescence characterization of Mg.2'(3')-O (N-methylanthraniloyl) nucleotides bound to the Dictyostelium discoideum myosin motor domain. AB - Mant (2'(3')-O-(N-methylanthraniloyl)) labeled nucleotides have proven to be useful tools in the study of the kinetic mechanism of the myosin ATPase by fluorescence spectroscopy. The sensitivity of the mant fluorophore to its local environment also makes it suitable to investigate the exposure of bound nucleotides to solvent from collisional quenching measurements. Here we present the crystal structure of mant-ADP and beryllium fluoride complexed with Dictyostelium discoideum myosin motor domain (S1dC) at 1.9 A resolution. We complement the structural approach with an investigation of the accessibility of the mant moiety to solvent using acrylamide quenching of fluorescence emission. In contrast to rabbit skeletal myosin subfragment 1, where the mant group is protected from acrylamide (Ksv=0.2 M-1), the fluorophore is relatively exposed when bound to Dictyostelium myosin motor domain (Ksv= 1.4 M-1). Differences between the Dictyostelium structure and that of vertebrate skeletal subfragment 1, in the region of the nucleotide binding pocket, are proposed as an explanation for the differences observed in the solvent accessibility of complexed mant nucleotides. We conclude that protection of the mant group from acrylamide quenching does not report on overall closure of the nucleotide binding pocket but reflects more local structural changes. PMID- 9405149 TI - Unique structural features of the monomeric Cu,Zn superoxide dismutase from Escherichia coli, revealed by X-ray crystallography. AB - The first three-dimensional structure of a functional monomeric Cu, Zn superoxide dismutase (from Escherichia coli, E_SOD) is reported at 2.0 A resolution (R factor=16.8%). Compared to the homologous eukaryotic enzymes, E_SOD displays a perturbed antiparallel beta-barrel structure. The most striking structural features observed include extended amino acid insertions in the surface 1, 2-loop and S-S subloop, modification of the disulfide bridge connection, and loss of functional electrostatic residues, suggesting a modified control of substrate steering toward the catalytic center. The active site Cu2+ displays a distorted coordination sphere due to an unusually long bond to the metal-bridging residue His61. Inspection of the crystal packing does not show regions of extended contact indicative of a dimeric assembly. The molecular surface region involved in subunit dimerization in eukaryotic superoxide dismutases is structurally altered in E_SOD and displays a net polar nature. PMID- 9405150 TI - Use of a 3D structure data base for understanding sequence-dependent conformational aspects of DNA. AB - The roll-twist-slide correlation in the DNA crystal structures that are collected in the Nucleic Acid Data Base is analyzed in order to obtain a general understanding of the effects of the nucleotide sequence on the 3D structure of a dinucleotide step. It is concluded that the differences between the pyrimidine bases and the purine bases in terms of their physical shapes are the major factors that determine the stereochemical characteristics of the steps through base to backbone and base to base interactions. The characteristics are further modulated by the differences between the A:T and G:C base-pairs, which can be explained by enhancement of the purine-pyrimidine asymmetry in the A:T base-pair. PMID- 9405151 TI - Structural study of the response regulator HupR from Rhodobacter capsulatus. Electron microscopy of two-dimensional crystals on a nickel-chelating lipid. AB - Two-dimensional crystals of the histidine-tagged-HupR protein, a transcriptional regulator from the photosynthetic bacterium Rhodobacter capsulatus, were obtained upon specific interaction with a Ni2+-chelated lipid monolayer. HupR is a response regulator of the NtrC family; it activates the transcription of the structural genes, hupSLC, of the [NiFe]hydrogenase. The lipid (Ni-NTA-DOGA) uses the metal chelator nitrilotriacetic group as the hydrophilic headgroup and contains unsaturated oleyl tails to provide the fluidity necessary for two dimensional protein crystallization. A projection map of the full-length protein at 18 A resolution was generated by analysing electron microscopy micrographs of negatively stained crystals. The HupR protein appeared to be dimeric and revealed a characteristic "propeller-like" motif. Each monomer forms an L-shaped structure. PMID- 9405152 TI - Interactions between HIV Rev and nuclear import and export factors: the Rev nuclear localisation signal mediates specific binding to human importin-beta. AB - The human immunodeficiency virus type 1 (HIV-1) Rev protein binds to unspliced HIV-1 pre-mRNA and exports it from the nucleus. Rev itself can "shuttle" between the nucleus and cytoplasm. This bi-directional transport is mediated by two specific Rev sequences: a nuclear localisation signal (NLS), which overlaps the RNA-binding domain, and a distinct nuclear export signal (NES). In this study we characterised new monoclonal antibodies that bind different epitopes of Rev, including the import and export sequences. In RNA bandshift assays, we observed that formation of a multimeric complex between Rev and its target RNA completely masks the Rev NLS, whereas the NES remains readily accessible. We then tested for signal-mediated interactions between Rev and different nuclear transport receptors, using mutations in the Rev NES or NLS to control for specificity. Extensive biochemical analyses did not reveal any direct NES-dependent interaction between Rev (free or RNA-bound) and the previously proposed export co factors, human RIP/Rab and eIF-5A. By contrast, similar tests showed that Rev binds directly via its arginine-rich NLS to the human nuclear import receptor, importin-beta. This interaction was highly specific and was abolished by mutation in the Rev NLS. Importin-beta did not bind to the RNA-bound form of Rev, providing a mechanism to ensure that Rev is imported only following release of its RNA cargo. Unlike many NLS-containing proteins that bind stably to an importin-alpha/beta heterodimer, the binding of Rev to importin-beta was actually blocked by importin-alpha receptor. Our findings suggest that Rev and importin alpha bind (via an arginine-rich sequence) to a similar region on importin-beta. In addition, we show that the complex between Rev and importin-beta can be dissociated by the nuclear Ran GTPase, but only when Ran is in the GTP-bound form. The series of interactions we describe provide a novel pathway for the import of Rev across the nuclear pore complex, and a mechanism for its release into the nucleoplasm. PMID- 9405153 TI - Coupled-enzymatic assays for the rate and mechanism of DNA site exposure in a nucleosome. AB - The packaging of DNA in nucleosomes presents obstacles to the action of gene regulatory proteins and polymerases on their natural chromatin substrates. We recently reported that nucleosomes exist in a conformational equilibrium, transiently exposing stretches of their DNA off the histone surface. Such "site exposure" processes potentially provide the needed access of proteins to DNA in chromatin. However, the experiments that reveal site exposure are carried out on timescales of tens of minutes to hours. The actual rates of site exposure are not known. Here we use T7 RNA polymerase and exonuclease III as probes to obtain a more relevant lower bound on the rate of nucleosomal site exposure. We find that the organization of DNA into nucleosomes detectably slows the elongation rate of the polymerase, but that full-length elongation, which requires access to all of the DNA, occurs on the seconds timescale. Independent experiments with exonuclease III, which probes the outermost DNA segments only, similarly show that site exposure in these regions occurs on a timescale of seconds or faster. We conclude that site exposure is sufficiently rapid that it may play a role in the initial binding of regulatory proteins to nucleosomal target sites. These rapid rates argue against a nucleosome sliding model for the mechanism of site exposure. Surprisingly, the measured rates may be too slow to account for the known rates of polymerase elongation in vivo. Mechanisms by which polymerase progression through nucleosomes might be catalyzed are discussed. PMID- 9405154 TI - Temperature-sensitive mutants of the EcoRI endonuclease. AB - The EcoRI endonuclease is an important recombinant DNA tool and a paradigm of sequence-specific DNA-protein interactions. We have isolated temperature sensitive (TS) EcoRI endonuclease mutants (R56Q, G78D, P90S, V97I, R105K, M157I, C218Y, A235E, M255I, T261I and L263F) and characterized activity in vivo and in vitro. Although the majority were TS for function in vivo, all of the mutant enzymes were stably expressed and largely soluble at both 30 degrees C and 42 degrees C in vivo and none of the mutants was found to be TS in vitro. These findings suggest that these mutations may affect folding of the enzyme at elevated temperature in vivo. Both non-conservative and conservative substitutions occurred but were not correlated with severity of the mutation. Of the 12 residues identified, 11 are conserved between EcoRI and the isoschizomer RsrI (which shares 50% identity), a further indication that these residues are critical for EcoRI structure and function. Inspection of the 2.8 A resolution X ray crystal structure of the wild-type EcoRI endonuclease-DNA complex revealed that: (1) the TS mutations cluster in one half of the globular enzyme; (2) several of the substituted residues interact with each other; (3) most mutations would be predicted to disrupt local structures; (4) two mutations may affect the dimer interface (G78D and A235E); (5) one mutation (P90S) occurred in a residue that is part of, or immediately adjacent to, the EcoRI active site and which is conserved in the distantly related EcoRV endonuclease. Finally, one class of mutants restricted phage in vivo and was active in vitro, whereas a second class did not restrict and was inactive in vitro. The two classes of mutants may differ in kinetic properties or cleavage mechanism. In summary, these mutations provide insights into EcoRI structure and function, and complement previous genetic, biochemical, and structural analyses. PMID- 9405155 TI - Resistance to nevirapine of HIV-1 reverse transcriptase mutants: loss of stabilizing interactions and thermodynamic or steric barriers are induced by different single amino acid substitutions. AB - The kinetic parameters governing the inhibition by Nevirapine of the RNA dependent DNA synthesis catalyzed by HIV-1 reverse transcriptase have been determined by steady-state kinetic analysis with the wild-type enzyme and with mutant reverse transcriptases containing the single amino acid substitutions L100I, K103N, V106A, V179D, Y181I and Y188L. While the mutant V179D was inhibited by Nevirapine as the wild-type enzyme, all the other mutations displayed a 17 to 90-fold reduced sensitivity to the drug in the order: Y181I<(i.e. less sensitive) Y188L < V106A < L100I < K103N < wild-type. Determination of the rate constants for Nevirapine binding (kon) and dissociation (koff) for the mutant and wild-type enzymes showed that mutations L100I and V106A increased the koff values by 12 and 8.5-fold, respectively, without significantly affecting the kon, whereas mutation K103N decreased the kon 5-fold without increasing the koff. Mutations Y181I and Y188L, on the other hand, conferred resistance to Nevirapine affecting both koff and kon values. In addition, mutations L100I and Y181I reduced the catalytic potential of HIV-1 RT. Thus, Nevirapine resistance could arise from a combination of loss of stabilizing interactions and emergence of steric and thermodynamic barriers for drug binding, depending on the particular amino acid substitution involved. PMID- 9405156 TI - Use of organic cosmotropic solutes to crystallize flexible proteins: application to T7 RNA polymerase and its complex with the inhibitor T7 lysozyme. AB - We have crystallized, using several approaches that may be of general interest, T7 RNA polymerase (T7RP) and the T7 RNA polymerase-T7 lysozyme complex (T7RPL) in forms suitable for structure determination by X-ray crystallography. A series of polyhydric alcohols, sugars, amino and methylamino acids, compounds known to stabilize protein structure, were found to be critical for both crystallization and subsequent improvement of the crystal's diffraction resolution. Moreover, optimal crystallogenesis was achieved through an unconventional "reverse" vapor diffusion sitting drop method that is suitable for proteins that are insoluble at low ionic strength.T7RP has been crystallized in an orthorhombic form (I), space group P222, with cell parameters a=220 A, b=205 A, c=67 A and a monoclinic form (II), space group P21, with cell parameters a=229 A, b=205 A, c=70 A, beta=106 degrees. Crystal form I diffracts X-rays to 3.5 A and form II to 6.0 A. Three and six copies of the polymerase are predicted to be in the asymmetric unit forms I and II, respectively. Three monoclinic crystal forms of the T7RPL complex have been obtained in space group C2. Form I has cell parameters a=320 A, b=93 A, c=229 A, beta=129 degrees, form II has parameters a=293 A, b=93 A, c=68 A, beta=93 degrees, and form III has parameters a=270 A, b=93 A, c=63 A, beta=103 degrees. Crystal form I diffracts synchrotron wiggler radiation to 3.2 A and form III to 2.8 A. Calculations of crystal density imply three or four copies of the complex in form I and one copy in the asymmetric unit of forms II and III. PMID- 9405157 TI - The 2.35 A crystal structure of the inactivated form of chicken Src: a dynamic molecule with multiple regulatory interactions. AB - The Src protein tyrosine kinase plays a critical role in a variety of signal transduction pathways. Strict regulation of its activity is necessary for proper signalling. We present here the crystal structure of chicken Src which is phosphorylated at Tyr527 and represents its least active form. Our structure, similar to the recently reported human Hck and Src structures, contains the SH3, SH2 and the kinase domains and the C-terminal regulatory tail but not the N terminal unique domain. The SH3 domain uses its hydrophobic surface to coordinate the SH2-kinase linker such that residues Gln251 and Leu255 specifically interact with side chains in the beta2-beta3 and the alphaC-beta4 loops of the N-terminal lobe opposite of the kinase active site. This position of the SH3 domain and the coordination of the SH2-kinase linker also optimally places the SH2 domain such that the phosphorylated Tyr527 in the C-terminal tail interacts with the SH2 binding pocket. Analogous to Cdk2 kinase, the position of the Src alphaC-helix in the N-terminal lobe is swung out disrupting the position of the active site residues. Superposition of other protein kinases including human Hck and Src onto chicken Src indicate that the alphaC-helix position is affected by the relative position of the N-terminal lobe with respect to the C-terminal lobe of the kinase and that the presence of the SH3/SH2-kinase linker/N-terminal lobe interactions restricts the kinase lobes and alphaC-helix access to the active conformation. These superpositions also suggest that the highly conserved alphaC-beta4 loop restricts the conformational freedom of the N-terminal lobe by anchoring it to the C-terminal lobe. Finally, based on sequence alignments and conservation of hydrophobic residues in the Src SH2-kinase linker as well as in the alphaC-beta4 and beta2-beta3 loops, we propose that the Src-related kinases, Abl, Btk and Csk, share the same quaternary structure. PMID- 9405158 TI - Binding of biotin to streptavidin stabilizes intersubunit salt bridges between Asp61 and His87 at low pH. AB - The remarkable stability of the streptavidin tetramer towards subunit dissociation becomes even greater upon binding of biotin. At two equivalent extensive monomer-monomer interfaces, monomers tightly associate into dimers that in turn associate into the tetramer at a less extensive dimer-dimer interface. To probe the structural basis for the enhancement of the stability of streptavidin by biotin, the crystal structures of apostreptavidin and its complexes with biotin and other small molecule and cyclic peptide ligands were determined and compared at resolutions as high as 1.36 A over a range of pH values from as low as 1.39. At low pH dramatic changes occur in the conformation and intersubunit hydrogen bonds involving the loop comprising Asp61 to Ser69. The hydrogen-bonded salt bridge between Asp61 Odelta2 and His87 Ndelta1, observed at higher pH, is replaced with a strong hydrogen bond between Asp61 Odelta1 and Asn85 Odelta1. Through crystallography at multiple pH values, the pH where this conformational change occurs, and thus the pKa of Asp61, was determined in crystals of space group I222 and/or I4122 of apostreptavidin and complexes. A range in pKa values for Asp61 was observed in these structures, the lowest being 1.78+/-0.19 for I222 streptavidin-biotin in 2.9 M (NH4)2SO4. At low pH the decrease in pKa of Asp61 and preservation of the intersubunit Asp61 Odelta2-Ndelta1 His87 hydrogen-bonded salt bridge in streptavidin-biotin versus apostreptavidin or streptavidin-peptide complexes is associated with an ordering of the flexible flap comprising residues Ala46 to Glu51, that in turn orders the Arg84 side-chain of a neighboring loop through resulting hydrogen bonds. Ordering of Arg84 in close proximity to the strong intersubunit interface appears to stabilize the conformation associated with the Asp61 Odelta2-Ndelta1 His87 hydrogen-bonded salt bridge. Thus, in addition to the established role of biotin in tetramer stabilization by direct mediation of intersubunit interactions at the weak interface through contact with Trp120, biotin may enhance tetramer stability at the strong interface more indirectly by ordering loop residues. PMID- 9405159 TI - Solution structure of human p8MTCP1, a cysteine-rich protein encoded by the MTCP1 oncogene, reveals a new alpha-helical assembly motif. AB - MTCP1 (for Mature-T-Cell Proliferation) is the first gene unequivocally identified in the group of uncommon leukemias with a mature phenotype. The three dimensional solution structure of the human p8(MTCP1) protein encoded by the MTCP1 oncogene was determined by homonuclear proton two-dimensional NMR methods at 600 MHz. After sequence specific assignments, a total of 931 distance restraints and 57 dihedral restraints were collected. The location of the three previously unassigned disulfide bridges was determined from preliminary DIANA structures, using a statistical analysis of intercystinyl distances. The solution structure of p8(MTCP1) is presented as a set of 30 DIANA structures, further refined by restrained molecular dynamics using a simulated annealing protocol with the AMBER force field. The r.m.s.d. values with respect to the mean structure for the backbone and all heavy atoms for a family of 30 structures are 0.73(+/-0.28) and 1.17(+/-0.23) A, when the structured core of the protein (residues 5 to 63) is considered. The solution structure of p8(MTCP1) reveals an original scaffold consisting of three alpha helices, associated with a new cysteine motif. Two of the helices are covalently paired by two disulfide bridges, forming an alpha-hairpin which resembles an antiparallel coiled-coil. The third helix is oriented roughly parallel to the plane defined by the alpha antiparallel motif and its axis forms an angle of approximately 60 degrees with respect to the main axis of this motif. PMID- 9405160 TI - Conformational flexibility and polymerization of vesicular stomatitis virus matrix protein. AB - The matrix protein of vesicular stomatitis virus (VSV) plays a pivotal role in viral assembly. We previously demonstrated the ability of M protein to self associate at low salt concentrations. Now, we show the ability of M protein to polymerize in the presence of ZnCl2 in a nucleation-dependent manner. Analysis of kinetics revealed that the nuclei are probably made of three or four molecules of M. These results are consistent with the idea that in vitro self association of M protein is not due to amorphous aggregation but rather reflects an intrinsic ability of M to polymerize. Using attenuated total reflectance Fourier transform infrared spectroscopy, we showed that M polymerization is associated with an increase in the beta-sheet content of the protein. We propose a model explaining both the apparent M protein solubility in infected cells and how M polymerization could promote viral assembly. Data available for other negative strand viruses suggest that M polymerization may be the general basis of viral assembly. PMID- 9405161 TI - Carl Ludwig, the Leipzig Physiological Institute, and introduction to the focused issue: growth factors and cardiac hypertrophy. PMID- 9405162 TI - Phosphatidic acid: a potential signal transducer for cardiac hypertrophy. AB - Phosphatidic acid (PA) is mainly formed by the hydrolysis of phosphatidylcholine due to the activation of phospholipase D (PLD). PA is also generated by phosphorylation of diacylglycerol (DAG) due to the action of DAG kinase and is converted to DAG under the action of PA phosphohydrolase. Most of the positive inotropic agents which are known to stimulate cardiac hypertrophy, have been shown to increase the level of PA in cardiac sarcolemma. Although the growth factor-like effect of PA has been recognized in a wide variety of tissues, there is a lack of similar information in adult cardiomyocytes. By using single cardiomyocytes, we have now shown that PA increased the basal [Ca2+]i level without significant effect on the amplitude of Ca2+ transients. PA (10-50 mu M) also increased the [Ca2+]i in cardiac cell suspension. PA has also been shown to stimulate protein synthesis in cardiomyocytes, which is inhibited by a PKC inhibitor as well as a Ca2+ chelator. PA at the concentration of 1-50 mu M was observed to stimulate the activity of PLC in cardiac sarcolemma; this effect was attenuated by a PLC inhibitor. Since DAG, formed due to the activation of PLC, is considered to play a crucial role in regulating the activity of protein kinase C (PKC), the positive feedback effect of PA on this pathway may be essential for maintaining the sustained elevation in the activity of PKC during the development of cardiac hypertrophy. In view of these observations and other facts available in the literature, it is suggested that PA may be a potential signal transducer for the development of cardiac hypertrophy. PMID- 9405163 TI - Signaling pathways in cardiac myocyte hypertrophy. AB - When a heart responds to increased workload it does so by hypertrophy. This is characterized by an increase in cell size in the absence of cell division, and is accompanied by distinct qualitative and quantitative changes in gene expression. The use of cardiomyocytes in cell culture has identified, besides mechanical loading, a range of substances, such as cytokines, growth factors, catecholamines, vasoactive peptides and hormones, involved in mediating cardiac myocyte hypertrophy, and has enabled the molecular dissection of the pathways involved in signal transduction. Many different pathways are activated in response to different hypertrophic stimuli, and a growing number of crosslinks are being characterized between these pathways. Recent evidence suggests a central role for Ras in transmitting signals from G-protein coupled receptors, from growth factor receptors and from cytokine receptors not only down the Raf MEK-ERK pathway to the nucleus, but also to various other cytosolic effectors. The evaluation of distinct morphological phenotypes, together with biochemical data on gene regulation, suggests that interactions between different signaling pathways take place. Each stimulus provokes a typical cellular phenotype and different stimuli may act alone or in concert in a synergistic, antagonistic or permissive manner. Consequently, hypertrophy of cultured cardiomyocytes cannot simply be characterized as the reversal to the fetal gene expression program. Thus, hypertrophic growth of the heart may similarly be the result of a complex combinatorial action of various stimuli, which may also lead to different morphological and biochemical phenotypes with distinct physiological properties. PMID- 9405164 TI - Molecular mechanisms of angiotensin II in modulating cardiac function: intracardiac effects and signal transduction pathways. AB - Angiotensin II (Ang II), the effector peptide of the renin-angiotensin system (RAS), regulates volume and electrolyte homeostasis and is involved in cardiac and vascular cellular growth in humans and other species. This system, which has been conserved throughout evolution, plays an important role in cardiac and vascular pathology associated with hypertension, coronary heart disease, myocarditis and congestive heart failure. The traditional RAS is viewed as a system in which circulating Ang II is delivered to target organs and cells. However, in the past decade, a local RAS has been described in cardiac cells, providing evidence for autocrine and paracrine pathways by which biological actions of Ang II could be mediated. The critical actions of Ang II are mediated primarily through the AT1, G-protein (guanylyl nucleotide binding protein) coupled receptor. In addition to coupling to conventional G-protein signal transduction pathways, the AT1 receptor was recently shown to increase the tyrosine phosphorylation of several intracellular substrates, including the STAT (Signal Transducers and Activators of Transcription) family of novel transcription factors, in rat cardiac fibroblasts, myocytes and vascular smooth muscle cells, and AT1 receptor transfected CHO cells. It has been shown that Ang II stimulates the tyrosine phosphorylation and nuclear translocation of Stat1 (Stat 91) and Stat3 (Stat 92). Angiotensin II acting directly through the AT1 receptor, induces the formation of a complex of STAT proteins termed SIF (sis inducing factor) which binds the DNA sequence, SIE (sis-inducing element) present in the promotor element of many genes. This provides evidence for a direct role of Ang II in mediating inflammatory and remodeling responses through the JAK-STAT pathway. Thus, it is likely that the JAK-STAT pathway has an important role in Ang II-mediated effects on gene transcription, cardiac and vascular cellular growth/development, and inflammatory responses. PMID- 9405165 TI - Correlation between function and proto-oncogene expression in isolated working rat hearts under various overload conditions. AB - In isolated perfused working rat hearts we have studied the effects of norepinephrine (NE) and of pressure as well as volume overload alone, and in combination on heart function and expression of the proto-oncogenes c-fos and c myc. This preparation was functionally stable over the observation period of 120 min. NE (3x10(-8) M) induced an immediate and sustained increase in aortic and coronary flow as well as in cardiac output. Volume overload was established by increasing left atrial filling pressure (preload) from 8-16 cm H2O and resulted in a marked increase in aortic flow and cardiac output which subsided somewhat over time. Pressure overload was created by elevation of afterload from 80-100 cm H2O and induced a decrease in aortic flow and a slight increase in coronary flow. Using specific cDNA clones, the mRNAs of c-fos and c-myc were measured by Northern blots and quantified with densitometry. In all three conditions, c-fos mRNA was increased first, after 30 min. It was more pronounced and of longer duration in the NE-stimulated hearts. The increase in c-myc-mRNA occurred after 60 or 90 min. After 120 min, all signals were normal, although heart function was still altered in a manner specific for the respective intervention. Combination of NE with preload and afterload elevation as well as combination of preload and afterload elevation led first to an increase in cardiac output which was followed by a decline to various degrees. In all these combinations, the c-fos mRNA signal appeared earlier, after 15 min, and persisted for a longer period of time compared with the effect of a single stimulus. The c-myc mRNA was increased later, after 30 or 60 min. This increase persisted throughout the entire observation period, showing a further progressive rise. These results show that stimuli which cause cardiac hypertrophy in vivo induce a transient and sequential increase in proto-oncogene expression in the isolated working rat heart. Combination of two hypertrophy-inducing stimuli elicit an earlier, more pronounced and longer-lasting expression of the proto-oncogenes c-fos and c-myc, while functional parameters deteriorate. PMID- 9405166 TI - Tumor necrosis factor alpha (TNF alpha) is cardiodepressant in pathophysiologically relevant concentrations without inducing inducible nitric oxide-(NO)-synthase (iNOS) or triggering serious cytotoxicity. AB - Cardiac hypertrophy and heart failure are frequently accompanied by elevated plasma levels of tumor necrosis factor alpha (TNF alpha), the pathogenetic relevance of this finding being a matter of debate. In human acute septic cardiomyopathy, on the other hand, the negative inotropic impact of TNF alpha on the heart is well documented and frequently ascribed to the induction of inducible nitric oxide (NO) synthase (iNOS) and an enhanced production of NO in the heart. Yet the present study presents evidence that in cardiomyocytes TNF alpha in non-toxic concentrations specifically depresses contractile performance independent of NO. In spontaneously beating neonatal rat cardiomyocytes, TNF alpha in a low, pathophysiologically relevant concentration (10 U/ml, 1-3 days) does not alter basal pulsation amplitude, but blocks alpha- and beta-adrenoceptor stimulated increase in contractility and beating irregularity and impairs the impact of high extracellular calcium on contractile performance. However, this low TNF alpha-concentration does not suffice to induce iNOS - documented by reverse transcriptase polymerase chain reaction - or enhance nitrite concentrations in the cell culture supernatants as a measure of cellular NO production, neither in the presence nor absence of dexamethasone (0.1 micro M). Only in high concentration - the specific proinflammatory action being documented by an enhanced release of interleukin-6 from cardiomyocytes - TNF alpha (1000 U/mol; 6, 24 h) weakly induces the mRNA for iNOS, with a consecutive moderate rise in cellular nitrite production. TNF alpha-incubation (10-1000 U/ml) does not alter the morphological appearance of the cells displayed by phase contrast microscopy or evoke gross cytotoxicity. PMID- 9405167 TI - Angiotensin receptors in cardiac and renal hypertrophy in rats. AB - Angiotensin II mediates its effects through activation of specific angiotensin (AT) receptors which can be regulated during cardiovascular disease. This study has investigated whether an increased cardiac and renal AT receptor density is important in the development of left ventricular and renal hypertrophy in three rat models of hypertension [spontaneous hypertensive (SHR), deoxycorticosterone acetate (DOCA)-salt and 2K1C renal hypertensive rats]. Although all hypertensive rats developed left ventricular and renal hypertrophy, AT receptor density increased only in the left ventricle and kidney of SHR during the development of hypertension. Thus, cardiac and renal hypertrophy per se do not increase AT receptor density. AT receptors were increased in the liver of DOCA-salt rats, 2K1C rats and 52-week-old SHR and in adrenal glands of DOCA-salt rats and SHR. A plausible explanation for tissue-dependent AT receptor regulation involves tissue selective control of local renin-angiotensin systems independent of circulating hormone levels, combined with disease-induced cell damage. PMID- 9405168 TI - Effects of angiotensin II receptor blockade on hypoxia-induced right ventricular hypertrophy in rats. AB - It was the aim of the present study to characterize the hemodynamic, biochemical and morphologic effect of angiotensin II receptor blockade on hypoxia-induced right ventricular hypertrophy in rats. Isolated right ventricular hypertrophy was induced in female Sprague-Dawley rats by intermittent hypoxia (IH; 10% O2, 8 h/day, 5 days/week, 20 days of exposition, n=15). After completion of IH, left- (LV) and right-ventricular (RV) hemodynamic parameters were measured under room air conditions in the intact, thiopental-anesthetized animals with special Millar ultraminiature tipcatheter-manometers. Cardiac output was determined using the thermodilution method. Cell volume (CV) of isolated cardiomyocytes was measured with a Coulter Channellyzer after collagenase cell isolation. The specific activities of the myocardial pentose phosphate pathway enzymes glucose-6 phosphate-dehydrogenase (G-6-PD) and 6-phosphogluconate-dehydrogenase (6-PGD) were determined using a spectrophotometric assay. IH caused a rise in right ventricular systolic pressure (RVSP) from 38.1+/-0.83 to 58.1+/-1.42 mmHg and an increase in the RV weight/body weight ratio (RVW/BW) from 0.884+/-0. 053 to 1.166+/-0.049 mg/g. The activities of G-6-PD and 6-PGD were significantly increased after IH in the RV, but not in the LV. CV was increased from 24 248+/ 1193 to 29 541+/-1765 micrometer 3, myocardial cell length was unchanged. IH had no influence on the LV parameters or cardiac output. Co-infusion of the angiotensin II receptor antagonist losartan (LO; 12 mg/kg/d i.p., n=14) during the IH period reduced the rises in RVSP (49.4+/-2.06 mmHg), RVW/BW (0. 99+/-0.072 mg/g), G-6-PD and 6-PGD significantly, but not completely. The increase in CV, however, was prevented (24 524+/-2370 micrometer 3) entirely. We conclude from these data that the IH-induced RV-hypertrophy was primarily of the concentric type. LO attenuated the hypoxia-induced isolated RV hypertrophy and significantly reduced the metabolic response of the RV. The LO effect was most potent with regard to the increase in cardiomyocyte volume. PMID- 9405169 TI - Contractile function of papillary muscle from rats with different infarct size after beta-adrenergic blockade and ACE-inhibition. AB - We tested whether ACE-inhibition with ramipril (A), beta-adrenergic blockade with metoprolol (beta) or combined treatment (beta A) for 6 weeks after inducing myocardial infarction in rats by left coronary artery ligation modifies contractile function of hypertrophied papillary muscle from left ventricles with different infarct size (IS) compared to a placebo group (P). At IS<40% of left ventricle, contraction and relaxation were less impaired than at IS>40% compared to sham operated rats (SO). Isometrically developed peak force and calcium sensitivity of myofilaments, measured in skinned fibres, were significantly higher in beta. Treatment with ramipril or metoprolol improved contraction rate and force development, respectively, mainly at IS<40%, but deteriorated relaxation rate. ACE-inhibition and beta-adrenergic blockade had no significant improving effect on the relaxation rate and further characteristics of the contractile function at IS>40%, although combined treatment reduced the infarct size and ramipril treatment suppressed the development of hypertrophy. Post extrastimulatory potentiation was increased in beta and beta A at IS>40%. Post rest potentiations were influenced hardly at IS<40% and were significantly smaller in A at IS>40%. The twitch-to-twitch decay of the potentiations was faster at IS>40%. Increase in the degree of post-extrastimulatory potentiation, steeper twitch-dependent decay of the potentiations and loss of rest-dependent potentiation at IS>40% indicate relatively increased trans-sarcolemmal Ca2+ transports via Ca2+ channels and Na+/Ca2+ exchange, partly modified by ramipril and metoprolol. The results demonstrate that ACE-inhibition and beta-adrenergic blockade induce a dissociation between trophic effects and phenotypic effects on contractile function after chronic infarction. PMID- 9405170 TI - Effects of ACE-inhibition on redox status and expression of P-selectin of endothelial cells subjected to oxidative stress. AB - Redox stress during post-ischemic reperfusion may be the prime signal for processes leading to myocardial remodelling and hypertrophy. Nitric oxide (NO) is antioxidative, antiadhesive for neutrophils (PMN) and antiproliferative. Thus, enhancing endothelial production of NO, e.g. by inhibiting breakdown of endogenous bradykinin via angiotensin converting enzyme (ACE), could be beneficial. The effect of cilazaprilat (CILA, 10 micro M), an ACE inhibitor, on redox status, expression of the adhesion molecule P-selectin, and PMN adhesion under conditions of oxidative stress was investigated in cultured human umbilical vein endothelial cells (HUVECs). Incubation of the cells with H2O2 (0.1 and 1 mm) for 15 min served as oxidative stimulus. The intra- and extracellular concentrations of reduced and oxidized glutathione (GSH and GSSG) were measured by HPLC as indicators of endothelial redox status. Expression of P-selectin was measured by flow cytometry. Furthermore, firm leukocyte adhesion to HUVECs was assessed. In controls, the intracellular ratio GSH/GSSG averaged 47 and dropped to 30 after incubation with 0.1 mm H2O2. The ratio declined to 6.5 with 1 mm H2O2. CILA blocked the effects of 0.1 mm H2O2, but was ineffective against 1 mm peroxide. The extracellular ratio did not discriminate between 0.1 and 1 mm H2O2, falling from 18 to 1 in both situations. P-selectin expression rose from 100% (control) to 146% after 1 mm H2O2 without CILA, but only to 114% in the presence of CILA. PMN adhesion was enhanced from about 1600 PMN per microwell (control) to 4300/well by 1 mm H2O2. CILA had no significant effect on adhesion (3900 PMN/well). Exposure of HUVECs to 0.1 mm H2O2 affected neither P-selectin expression nor PMN adhesion. Consequently, ACE inhibition can mitigate mild (0.1 mm H2O2) but not more severe redox stress in HUVECs. Irrespectively, CILA reduced the upregulation of P-selectin at the higher H2O2 concentration, indicating that this process is regulated independently of the cellular redox status. The firm adhesion of PMN to HUVECs was independent of P-selectin expression. PMID- 9405171 TI - Different pathophysiology of cardiac hypertrophy in hypertension and hypertrophic cardiomyopathy. AB - In order to investigate differences in the pathophysiology of cardiac hypertrophy between patients with arterial hypertension and hypertrophic cardiomyopathy, DNA synthesis by cardiac myocytes and the effects of an angiotensin-converting enzyme inhibitor were examined in these two groups of patients. DNA synthesis and the cell cycle were investigated by flow cytometry using autopsy materials from patients with hypertension and hypertrophic cardiomyopathy. The hypertension group (n=10) included four men and six women aged 61+/-10 years (heart weight: 470+/-79 g, mean+/-s.d.); the cardiomyopathic group (n=10) included eight men and two women aged 61+/-23 years (heart weight: 615+/-211 g). The percentage of cells in G2M phase of the cell cycle was significantly decreased in the myocardium from patients with hypertrophic cardiomyopathy compared with that from hypertensive patients (cardiomyopathy v hypertension: 1.1+/-0.6 v 7.7+/-2.6%, mean+/-s.d.). Captopril, an angiotensin-converting enzyme inhibitor, was administered for 12 months to patients with hypertension (n=20) and hypertrophic cardiomyopathy (n=15). Regression of cardiac hypertrophy was assessed by echocardiography. Long term administration of captopril achieved regression of cardiac hypertrophy in the hypertensive patients, but not in the patients with hypertrophic cardiomyopathy. In the hypertensive patients, the left ventricular mass was 234+/ 9 g before treatment and 198+/-26 g after treatment (mean+/-s.d., P<0. 01). In cardiomyopathic patients, on the other hand, there was no significant difference of left ventricular mass after treatment (before v after, 305+/-85 v 285+/-90 g, mean+/-s.d.). These results suggest that the mechanism of cardiac hypertrophy differs between patients with hypertension and hypertrophic cardiomyopathy. PMID- 9405173 TI - Effects of hypoxia and metabolic inhibition on increases in intracellular Ca2+ concentration induced by Na+/Ca2+ exchange in isolated guinea-pig cardiac myocytes. AB - We have assessed if the sarcolemmal Na+/Ca2+ exchange of isolated cardiac myocytes becomes inhibited when the cells are subjected to hypoxia and metabolic inhibition. Function of the exchange was assessed by exposing single cardiac myocytes to Na+-free solutions and measuring the increases in [free Ca2+]i using the fluorescent indicator Fura-2. Removal of extracellular Na+ reverses the Na+ gradient and causes Ca2+ influx via Na+/Ca2+ exchange. This influx may trigger further Ca2+ release from the sarcoplasmic reticulum. Hypoxia and metabolic inhibition produced either by (1) vigorously bubbling glucose-free Tyrode with N2, or (2) bubbling with N2 and adding 10 mm 2-deoxy-D-glucose to the Tyrode or (3) using 0.1-1.0 mm sodium dithionite (sodium hyposulphite, Na2S2O4) in glucose free Tyrode caused a significant decline in the rates of increase in [free Ca2+]i during Na2+-free superfusion. The decreases in [free Ca2+]i on re-application of Na+ were also significantly slowed. This suggests that reverse-mode Na+/Ca2+ exchange and the processes leading to release of Ca2+ from the sarcoplasmic reticulum are inhibited in hypoxia and metabolic inhibition. PMID- 9405172 TI - Growth factor expression of human arterial smooth muscle cells and endothelial cells in a transfilter coculture system. AB - By releasing growth factors, vascular cells can modulate proliferation and migration of neighboring cells in the arterial wall. Previous histological studies in transfilter cocultures, a culture model aimed to simulate vessel wall architecture, indicated that human arterial endothelial cells (haEC) can influence human arterial smooth muscle cell (haSMC) growth significantly. The aim of this study was to investigate the expression and secretion of various growth factors in order to better define the functional interactions between haEC and haSMC. Protein levels of platelet-derived-growth factor-AB (PDGF-AB), transforming-growth factor-beta1 (TGF-beta1), and tumor-necrosis factor-alpha (TNF-alpha) in mono- and cocultures were determined by ELISA 6, 12, 24, 48, 72 h after serum reduction. Highest PDGF-AB levels were found in monocultures with proliferative haEC, showing a peak after 24 h. In cocultures of haEC and haSMC, PDGF-AB levels were significantly lower. In contrast, neither proliferative, nor confluent haSMC released PDGF-AB significantly. Highest TGF-beta1 concentrations were detected in cocultures, followed by monocultures of haSMC and monocultures of haEC. In all cultures, TGF-beta1 levels increased in parallel with cultivation time and cell numbers, showing a maximum after 72 h. TNF-alpha could not be detected in any culture. Northern blots demonstrated a strong expression of PDGF B chain-mRNA in haEC, but not in haSMC. PDGF-A chain and TGF-beta1-mRNA were expressed by haSMC and haEC. Addition of PDGF-AB to haSMC resulted in a potent growth stimulation, whereas TGF-beta1 and TNF-alpha exerted only moderate, divergent effects on haSMC. Histological observations in transfilter cocultures demonstrated that proliferative haEC induce the formation of fibromuscular plaques. These results suggest that proliferative haEC act as potent growth stimulators for haSMC, predominantly by PDGF-AB or -BB release. PMID- 9405174 TI - Cardiomyoplasty - improvement of muscle fibre type transformation by an anabolic steroid (metenolone). AB - Dynamic cardiomyoplasty, a method to support ventricular function by the chronically stimulated latissimus dorsi muscle wrapped around the heart is accompanied by a loss of mass and force of the transplanted muscle. These effects and the fast-to-slow transformation of the muscle could be possibly influenced by the additional administration of anabolic steroids. In this study, the left latissimus dorsi muscles of 12 sheep were electrically conditioned (group A). In 12 other animals (group B), stimulation was combined with the administration of metenolone (100 mg/week). Biopsies were taken from the right and left muscles at the beginning and after 6 and 12 weeks of treatment, frozen and cross-sectioned. The muscle fibre type composition was studied enzymhistochemically (SDH-staining and Myosin-ATPase-reaction) and immunocytochemically (using antibodies against different myosin heavy chains, MHC). Furthermore, the expression of different MHC isoforms was investigated electrophoretically. The untreated latissimus dorsi muscle contains 20% type I fibres expressing slow MHC and 80% type II fibres expressing fast MHC. After 6 weeks, the respective fibre type composition was 42 and 58% (group A) and 80 and 20% (group B). After 12 weeks, the percentage of the type I fibres rose in group A to 59% and in group B to 98%. In accordance with these morphological results, the MHC pattern determined electrophoretically showed a corresponding shift from the fast to the slow isoform. Therefore, the administration of metenolone avoids severe muscle atrophy, and improves and accelerates fast to slow fibre type conversion necessary for successful cardiomyoplasty. PMID- 9405175 TI - Repetitive stretching during low-flow ischemia impairs function in isolated porcine hearts: a model for segmental dyskinesis. AB - Myocardial infarction appears after 20 min of regional no-flow ischemia in vivo, but only after a much longer duration of global ischemia in isolated hearts. We tested whether repetitive myocardial stretching (RMS), as occurs in segmental ischemia, is involved in the pathogenesis of myocardial cell injury. Furthermore, we evaluated the role of stretch-activated channels by using Gadolinium (Gd3+). Isolated piglet hearts were perfused with red cell enriched Krebs-Henseleit buffer. RMS was induced by inflating a balloon in the left ventricle, using a control system to provide a pressure of 120 mmHg during one-third of the cycle and 0 mmHg during the rest of the cycle, with a frequency 150 per min. Function and metabolism were compared during 2 h of low-flow ischemia (10% of control), with and without RMS, followed by 1 h of reperfusion. Non-RMS hearts were exposed to saline (Isch), or Gd3+ 25 micromol/l (Gd3+-Isch). During ischemia, left ventricular systolic pressure (LVSP) stabilized in non-RMS hearts, but a further decrease, combined with increased anaerobic metabolism occurred in RMS hearts. After 30 min of reperfusion in the non-stretched hearts, LVSP returned to 77+/-4% of control (mean+/-s.e.) in the Isch group, and to 74+/-2% in the Gd3+-isch group (between groups; P=n.s.). In hearts exposed to RMS, LVSP returned to only 46+/-4% of control (RMS) and to 51+/-3% in the Gd3+-RMS group (both P=0.01 v Isch). The same alterations were seen for LV dP/dt. In RMS hearts, tissue concentrations of ATP were reduced and concentrations of lactate increased. We conclude that stretching of ischemic myocardium severely increases anaerobic metabolism and reduces functional and metabolic recovery. Blockade of stretch activated channels by Gd3+ does not prevent this effect. Thus, the reduced recovery induced by RMS is due to factors other than ion fluxes through stretch-activated channels. PMID- 9405176 TI - Comparison of in vitro preconditioning responses of isolated pig and rabbit cardiomyocytes: effects of a protein phosphatase inhibitor, fostriecin. AB - Calcium tolerant pig and rabbit cardiomyocytes were isolated using retrograde aortic perfusion of nominally calcium-free collagenase. Preconditioning protocols used 1 or 3x10-min episodes of ischemic pelleting or pre-incubation with 100 micro M adenosine, followed by a 15-min post-incubation and 180-240-min ischemic pelleting. Control cells were incubated and washed in parallel with the experimental groups. Injury was assessed by determination of cell morphology, trypan blue permeability following osmotic swelling, lactate and HPLC analysis of adenine nucleotides. Preconditioned pig cardiomyocytes had a reduced rate of ischemic contracture, but protection occurred without conservation of ATP. Preconditioned rabbit cardiomyocytes were protected without significant changes in rates of ischemic contracture or ATP depletion. Incubation of ischemic cells with the protein phosphatase inhibitor, fostriecin, at PP2A-selective concentrations (0.1-10 micro M), mimicked preconditioning in both rabbit and pig cardiomyocytes. In rabbits, the KATP channel blocker, 5-hydroxydecanoate (5-HD), did not block preconditioning or fostriecin protection. In the pig, 5-HD blocked both preconditioning and fostriecin protection, with return of the rates of ischemic contracture to control. However, 5-HD was an effective blocker of protection only in early ischemia. Fostriecin mimicked preconditioning in the rabbit and the early responses of the preconditioned pig. Preconditioning appears associated with protein phosphorylation in both the rabbit and the pig, but major pathways leading to protection may differ in the two species. PMID- 9405177 TI - Patterns of mobilization of copper and iron following myocardial ischemia: possible predictive criteria for tissue injury. AB - Direct evidence for substantial iron and copper mobilization into the coronary flow immediately following prolonged, but not short, cardiac ischemia is presented. When small volumes of coronary flow fractions (CFFs) were serially collected upon reperfusion, after 25-60 min of ischemia, the copper and iron levels in the first CFF were 50-fold and 12- to 15-fold higher, respectively, than corresponding pre-ischemic values. The copper and iron levels after shorter periods (15-21 min) of ischemia were only about five-fold higher than the pre ischemic values. This demonstrates that the resumption of coronary flow is accompanied by a burst of both metal ions. The levels of Cu/Fe in the CFFs correlated well with the loss of cardiac function following global ischemia of varying duration. After 18 min of ischemia, the residual cardiac function was less than 50%, and the damage was essentially reversible. After 25 min of ischemia, it exceeded 50% and was only partially reversible, while after 35 min, the damage exceeded 80%, and was mostly irreversible. The results are in accord with the hypothesis that copper and iron play causative roles in myocardial injury through mediation of hydroxyl radical production. Thus, the pattern of Cu/Fe mobilization from the tissue into the CFF can be used for the prediction of the severity of myocardial damage following ischemia, and could be developed into useful modalities for intervention in tissue injury. PMID- 9405178 TI - Distribution, splicing and glucocorticoid-induced expression of cardiac alpha 1C and alpha 1D voltage-gated Ca2+ channel mRNAs. AB - Dihydropyridine-sensitive voltage-gated (l-type) Ca2+ channels play an essential role in cardiac and smooth muscle excitation-contraction coupling. Transcripts for the two pore-forming subunits of l-type Ca2+ channels, alpha 1C and alpha 1D have been detected in heart and lung; however, distribution, structure and regulated expression of these channel subunit mRNAs have not been examined in detail. Here we use RNase protection and RT-PCR assays to identify cardiac specific features of expression of the two channel mRNAs. First, expression of alpha 1D mRNA is found in lung, aorta and atrium, but is not detected in ventricle. Limited expression of alpha 1D mRNA is also seen in enriched preparations of cardiac myocytes: it is significant in atrial myocytes, but not in ventricular myocytes. Second, RT-PCR analyses indicate that atrial alpha 1D channels exclusively contains the 15-amino acid insertion between third and fourth segments in repeat IV. Finally, expression of alpha 1C mRNA, but not alpha 1D mRNA, is up-regulated by glucocorticoids in atrium and ventricle: adrenalectomy and subsequent injection of the glucocorticoid agonist dexamethasone decreased and increased the channel message, respectively. Dexamethasone also significantly increased the number of dihydropyridine-binding sites in ventricle. In contrast, alpha 1C mRNA levels were glucocorticoid insensitive in lung and aorta. Thus, basal and glucocorticoid-induced expression, and splicing of the two l-type Ca2+ channel alpha 1 subunit transcripts are tissue specifically controlled in atria and ventricles of rat heart. PMID- 9405179 TI - Troponin T mRNA and protein isoforms in the human left ventricle: pattern of expression in failing and control hearts. AB - We and others have previously cloned several cDNAs of human cardiac troponin T (cTnT), demonstrating the multiplicity of cTnT isoforms in the human heart. Four of them named cTnT1, 2, 3 and 4 result from a combinatorial alternative inclusion of 30- and 15-nucleotides in the 5' coding region of the cDNAs. In failing human ventricles, increased expression of cTnT4 has been reported at the protein level. More recent RT-PCR experiments showed increased expression of fetal-type splicing products in the 5' region, one of them corresponding to cTnT1. To clarify this issue, we examined the accumulation of the 4 cTnT mRNA and protein species in left ventricular specimens at the time of heart transplantation, and in control left ventricular samples using RNase protection and Western blotting. In all samples, cTnT3 was the major mRNA isoform, cTnT4 a minor isoform while cTnT1 and cTnT2 mRNAs were present but barely detectable. At the protein level, cTnT3, 4 and 1 were detected with the same relative abundance as that seen at the mRNA level. In addition, we detected a fourth TnT species of very low abundance corresponding either to a skeletal or to a "short" cardiac TNT isoform. Compared to controls, increased levels of cTnT4 mRNA and protein were detected in only half the failing ventricles independently of the cause of failure, suggesting that this increase may not be a general characteristic of left ventricular failure but instead could be related to stress. Unexpectedly, we found a decrease in cTnT1 protein expression in all failing ventricular samples studied, compared to controls. PMID- 9405180 TI - Effects of PTH-rP(107-111) and PTH-rP(7-34) on adult cardiomyocytes. AB - We investigated whether parathyroid hormone-related peptide (PTH-rP), recently found expressed in the heart, exerts growth and contractile effects on adult cardiomyocytes from rat hearts. Synthetic PTH-rP peptides were used covering either a protein kinase C (PKC)-activating domain [PTH-rP(107-111)], or an adenylate cyclase activating domain [PTH-rP(1-34) and PTH-rP(7-34)]. PTH-rP(107 111) (1 micro M) increased creatine kinase BB activity (CK-BB), a CK isoform re expressed during cardiac hypertrophy, within 24 h by 62+/-12%. This induction was abolished in the presence of the mitogen-activated-protein (MAP)-kinase-kinase inhibitor PD 98059. PTH-rP(107-111) activated p42-MAP-kinase within 15 min, increased protein synthesis (19+/- 4%), total protein mass (19+/-5%), cell volume (45+/-7%), and cross-sectional area (38+/-9%) of cardiomyocytes. Activation of p42-MAP-kinase and increase in protein synthesis were abolished in presence of bisindolylmaleimide, a PKC inhibitor. PTH- rP(107-111) did not directly influence contractile activity but reduced the contractile response to isoprenaline. In contrast, PTH-rP(1-34) and PTH-rP(7-34) induced spontaneous contractile activity in 3-day-old cultures. This induction was abolished in presence of Rp-cAMPS, a protein kinase A inhibitor, indicating an involvement of cAMP in this response. PTH-rP(1-34) also increased the cellular accumulation of cAMP. It is concluded that PTH-rP exert direct effects on adult cardiomyocytes by activating either PKC via a functional domain covered by amino acids 107-111 or by activation of cAMP dependent protein kinase via a functional domain covered by amino acids 7-34. Since these parts of PTH-rP have either no homology [PTH-rP(107-111)] or only a limited structural similarity [PTH-rP(7-34)] to parathyroid hormone, these activities of PTH-rP have to be clearly distinguished from those described for parathyroid hormone. PMID- 9405181 TI - Preconditioning with ischaemia reduces both myocardial oxygen consumption and infarct size in a graded pattern. AB - We tested the hypothesis that ischaemic preconditioning reduces pre-ischaemic energy demand and thereby reduces the energy supply-demand mismatch imposed by coronary artery occlusion. Experiments were performed in 52 open chest pigs anaesthetised with sodium pentobarbital. One or two cycles, each of 10 min LAD occlusion followed by 30 min reperfusion, served as the preconditioning stimuli. The degree of protection was evaluated by measuring infarct size (tetrazolium stain) as percentage of area at risk (fluorescent particles) after 45 min LAD occlusion followed by 2 h reperfusion. One preconditioning cycle reduced regional myocardial oxygen consumption (MVO2) by 15+/-3% (P<0.05), whereas two cycles of preconditioning reduced MVO2 by 25+/-3% (P<0.05 v one cycle). This reduction was probably due to reduced energy demand, as both coronary blood flow and arteriovenous oxygen differences decreased, without lactate release or reduction in peak hyperaemic flow response. Energy requirements were most likely also reduced during ischaemia since repayment of flow debt after the second ischaemic period was 33+/-7% less than after the first ischaemic period (P<0.001). One preconditioning cycle reduced infarct size from 58+/-3% of area at risk to 40+/ 5% (P<0.05), whereas two cycles of preconditioning reduced infarct size to 15+/ 4% of area at risk (P<0.05 v one cycle). We conclude that preconditioning with ischaemia reduces energy consumption in a graded pattern. This effect may contribute to the graded protective effect of ischaemic preconditioning. PMID- 9405182 TI - Decrease in ischemic tolerance with aging in isolated perfused Fischer 344 rat hearts: relation to increases in intracellular Na+ after ischemia. AB - While the ischemic tolerance of the myocardium has been reported to decrease with senescence, it is not known when and how this occurs. Our objectives were to determine whether the tolerance to myocardial ischemia in rats decreased before the onset of senescence and whether an increase in myocardial ionic imbalance was associated with an enhanced myocardial injury with aging. Hearts were isolated from Fischer 344 rats categorized as young (12 weeks old), mature adult (24 weeks), middle-aged (50 weeks) or senescent (100 weeks). Hearts were perfused isovolumically by the Langendorff procedure and subjected to 25 min of global ischemia followed by 30 min of reperfusion. In the 50- and 100-week-old rats, the recovery of ventricular function and high-energy phosphate levels was lower and there was increased incidence of ventricular fibrillation after 25 min of global ischemia followed by reperfusion. The release of creatine kinase and lactate dehydrogenase during reperfusion was greater in the 50-and 100-week-old rats than in the 12- and 24-week-old rats, indicating the irreversible myocardial damage due to ischemia-reperfusion increased by middle-age. Intracellular levels of Na+ and K+ before ischemia were higher in the 50- or 100-week-old rats than in the 12 week-old rats. The increase in intracellular Na+ at end of ischemia was greater in the older (50-week-old, 215% of the pre-ischemic value; 100-week-old, 232% of the pre-ischemic value) than in the younger rats (12-week-old, 158% of the pre ischemic value). Results indicated that the rat heart becomes more vulnerable to ischemia in middle-age. This decrease in ischemic tolerance may be caused by an acceleration of myocardial ionic imbalance with aging. PMID- 9405183 TI - Association between hemodynamic parameters and the degeneration of sustained ventricular tachycardias into ventricular fibrillation in rats. AB - Sustained ventricular tachycardias (VT) often degenerate into ventricular fibrillation (VF). In the present study, the impact of VT on mean arterial blood pressure (MAP), myocardial blood flow (MBF), and myocardial oxygen consumption (MVCO2) was assessed. In addition, the degeneration of sustained VT into VF was analysed with respect to MAP. MBF was measured in 48 anesthetized rats with colored microspheres; arterial catecholamine levels were measured by HPLC in 16 additional rats during control conditions and VT. MBF (4. 66+/-1.29 ml/g/min; mean+/-s.d.) did not change with the onset of VT (5.37+/-1.92 ml/g/min, n.s.). Epinephrine (0.22+/-0.13 ng/ml) and norepinephrine (0.37+/-0.12 ng/ml) increased during VT (3.55+/-2.68 ng/ml, P<0.01; 0.88+/-0.44 ng/ml, P<0.05), respectively. VF was more frequent when MAP remained normal (MAP>80 mmHg: 26%) than with hypotension (MAP<80 mmHg: 2%, P<0.05). Mechanical failure was observed in 10% of rats with severe hypotension (MAP<60 mmHg), and 2% with moderate hypotension (MAP 60-80 mmHg). The endo-epicardial MBF ratio in the VF group was significantly lower than that in the non-VF group (0.94+/-0.17 v 1.11+/-0.24, P<0.05). CONCLUSIONS: severe hypotension predisposes to the occurrence of acute mechanical failure during VT; moderate hypotension during VT, however, serves as a protective mechanism against VF in structurally normal hearts. Subendocardial hypoperfusion in the presence of an increased energy demand during VT is suggested to be responsible for the initiation of VF. PMID- 9405184 TI - Secretion of cardiac plasminogen activator during hypoxia-induced right ventricular hypertrophy. AB - In the circulation, fibrinolytic activity is determined to a large degree by the relative levels of tissue plasminogen activator (tPA) and its major inhibitor (PAI-1). Vascular beds in different organs secrete tPA and PAI-1 into the circulation, and the total secretory rate of each protein is balanced by its half life in the bloodstream. We are testing the hypothesis that in the heart, ventricular hypertrophy will alter the rates of formation of tPA and/or PAI-1 and the rates of their release into the cardiac vasculature. In this study, we have examined the effects of continuous hypoxia on PA activity in extracts of rat heart ventricles, on the activity secreted into the cardiac vasculature of perfused hearts, and on the levels of mRNAs for tPA and PAI-1. Rats were subjected to hypobaric hypoxia at 0.5 atm for 1-21 days. The treatment caused polycythemia within 1-3 days, and right ventricular hypertrophy by 3 days. PA activity in extracts of both right and left ventricles was significantly elevated after 3 days of hypoxia, continued to increase for 4 additional days, and remained elevated for 3 weeks. The actions of inhibitors of urokinase and tPA indicated that the PA activity in heart extracts was exclusively tPA. Fibrin zymography confirmed that result. The mRNAs for tPA and for PAI-1 were elevated after 1 day of hypoxia and then returned to near control levels on days 2 and 3. After 7 days, hearts from hypoxic rats secreted more tPA activity into perfusates than did hearts from controls. The difference in secretory rates was proportional to the differences in the levels of tPA in the corresponding heart extracts. PMID- 9405185 TI - Study of microcirculation by coloured microspheres and NMR-microscopy in isolated rat heart: effect of ischaemia, endothelin-1 and endothelin-1 antagonist BQ 610. AB - Although the investigation of coronary microcirculation is of great importance, available methods have severe restrictions. They do not allow the study of vasodynamics of resistance vessels and microscopic conductance vessels simultaneously in the isolated beating rat heart. We now demonstrate that the combined measurement of perfusion which reflects the state of resistance vessels and cross-sections of microscopic conductance vessels is feasible in the model of the isolated constant flow perfused rat heart. Perfusion measurement was based on injection of coloured microspheres. Cross-sections of microscopic conductance vessels (diameter >140 micron) were determined by NMR-microscopy by flow weighted imaging. Both methods were established recently by our group. The combined measurement was applied to hearts which were subjected to ischaemia and reperfusion (group 1: n=5, 15 min ischaemia/group 2: n=7, 30 min ischaemia/measurements before ischaemia and 15/30 min after reperfusion), 200 pmol endothelin-1 bolus application (group 3: n=6/measurements before and 5 min after drug application), continuous infusion of the endothelin-1 antagonist BQ 610 (group 4: n=6/measurements before and 20 min after onset of infusion), and 200 pmol endothelin-1 application superimposed on 20 min of continuous BQ 610 infusion (group 5: n=7/combined measurement before BQ 610 infusion and 5 min after endothelin-1 application). In group 1, 15 min reperfusion restored the pre ischaemic perfusion state, whereas conductance vessels were dilated (80.8+/ 2.6%), after 30 min reperfusion pre-ischaemic conditions were also restored for conductance vessels. In group 2, a redistribution of perfusion from left ventricular endocardium to the right ventricular wall was observed. Post ischaemic rhythm disturbances made NMR-imaging in this group impossible. In group 3, a shift of perfusion from the left ventricular myocardium to the right ventricular wall was observed. Similarly, the cross-section of left ventricular conductance vessels decreased (-32.6+/-2.1%), whereas size of right ventricular vessels increased. In group 4, BQ 610 had no effect on perfusion nor on vessel size and antagonized the effect of endothelin-1 on perfusion and vessel size in group 5. PMID- 9405186 TI - Model systems for modulating the free energy of ATP hydrolysis in normoxically perfused rat hearts. AB - This study has two objectives; first, to develop perfusion conditions that decrease the free energy of ATP hydrolysis, Delta GATP, in isolated hearts; and, second, to modulate the Delta GATP in these perfused hear models. To accomplish the first goal, a series of inhibitors was employed to restrict acetyl-CoA oxidation. The second goal was accomplished by increasing work demand. Rat hearts were perfused with Krebs-Henseleit solution containing glucose and either; (i) no inhibitors (G group hearts); (ii) 0.3 mm bromo-octanoate (BrO), an inhibitor of beta-oxidation (GB group); (iii) 0.4 mm amino-oxyacetate (AOA), an inhibitor of the malate-aspartate shuttle (GA group); (iv) BrO and AOA (GBA group hearts); or (v) BrO, AOA, and 4 mm butyrate, an alternate substrate (GBA-Bu). Pacing hearts at 300 beats per min (beats/min), at 450 beats/min, and at 450 beats/min in the presence of 80 microgram/l dobutamine allowed three increasing levels of work demand to be attained. The Delta GATP values of the five groups of hearts were calculated for each workstate using the concentrations of high energy phosphate metabolites measured by 31P NMR spectroscopy. At the highest levels of workload demand, the G, GB, and GBA-Bu group hearts had Delta GATP values >/=-53 kJ/mol ATP. At the highest levels of workload demand, the GA and GBA hearts had Delta GATP values 20 min. The G, GB, and GBA-Bu hearts attained RPPs of >/=54x10(3) mmHg/min at the highest levels of workload demand. The GA and GBA hearts attained RPPs of 1% of transfected parasites). These vectors may-but need not-include mutant DHFR-TS alleles that confer pyrimethamine resistance to transgenic parasites. Large genomic constructs integrate at the endogenous locus by homologous recombination, but cDNA-derived sequences lacking long stretches of contiguous genomic DNA (due to intron excision) typically integrate into chromosomal DNA by nonhomologous recombination. Nonhomologous integration occurs effectively at random; and coupled with the high frequency of transformation, this allows a large fraction of the parasite genome to be tagged in a single electroporation cuvette. Genomic tagging permits insertional mutagenesis studies conceptually analogous to transposon mutagenesis in bacteria, yeast, Drosophila, etc. In theory (and, thus far, in practice), this allows identification of any gene whose inactivation is not lethal to the haploid tachyzoite form of T. gondii and for which a suitable selection or screen is available. Transformation vectors can be engineered to facilitate rescue of the tagged locus and to include a variety of reporters or selectable markers. Genetic strategies are also possible, using reporters whose function can be assayed by metabolic, visual, or immunological screens to "trap" genes that are activated (or inactivated) under various conditions of interest. PMID- 9405196 TI - Development of molecular genetics for Neospora caninum: A complementary system to Toxoplasma gondii. AB - The development of molecular genetics has greatly enhanced the study of Toxoplasma gondii, and investigations into the biology and pathology associated with neosporosis will be similarly benefited by the development of molecular tools for Neospora caninum. We have demonstrated the feasibility of using the existing DNA vectors developed for T. gondii to transfect and transform the Nc-1 strain of Neospora. We have also shown that T. gondii proteins are faithfully expressed and targeted in N. caninum, indicating the suitability of using Neospora as a heterologous expression system for studying T. gondii. These studies provide the basis for initiating molecular genetic studies on N. caninum and will allow for a number of molecular comparisons of these two closely related, though phenotypically distinct, parasites. Here we describe the methods and reagents used to perform genetic manipulations of N. caninum, and we present some of the principles and potential utilities of these molecular studies, including the use of N. caninum as a heterologous system for the study of T. gondii proteins. PMID- 9405197 TI - Transfection of malaria parasites. AB - The stable genetic transformation of three phylogenetically diverse species of Plasmodium, the parasitic etiological agent of malaria, is now possible. The parasite is haploid throughout the vast majority of its life cycle. Therefore with the single selectable marker activity and protocols currently available, it is possible not only to express introduced transgenes but also to study the effects of site-specific homologous recombination such as gene knockout. Transgene expression will allow the detailed study of many aspects of the cellular biology of malaria parasites, for example, the mechanisms underlying drug resistance and protein trafficking. We describe here the methods for propagation of the two animal models (Plasmodium berghei and Plasmodium knowlesi) and for transfection of these two species and the human parasite, Plasmodium falciparum. Examples of transgene expression are given. PMID- 9405198 TI - Gene targeting in malaria parasites. AB - Gene targeting, which permits alteration of a chosen gene in a predetermined way by homologous recombination, is an emerging technology in malaria research. Soon after the development of techniques for stable transformation of red blood cell stages of Plasmodium falciparum and Plasmodium berghei, genes of interest were disrupted in the two species. The main limitations of gene targeting in malaria parasites result from the intracellular growth and slow replication of these parasites. On the other hand, the technology is facilitated by the very high rate of homologous recombination following transformation with targeting constructs (approximately 100%). Here, we describe (i) the vector design and the type of mutation that may be generated in a target locus, (ii) the selection and screening strategies that can be used to identify clones with the desired modification, and (iii) the protocol that was used for disrupting the circumsporozoite protein (CS) and thrombospondin-related anonymous protein (TRAP) genes of P. berghei. PMID- 9405199 TI - Methods to prepare RNA and to isolate developmentally regulated genes from Eimeria. AB - Coccidians represent a large class of important intracellular parasites that traverse multiple developmental stages that are distinct and required to complete the life cycle. The biochemical details underlying the regulation of transformation from one developmental form to the next are limited and the study of such details presents unique obstacles. However, the genetic program is critical and may provide a basis for understanding the biology of these organisms in addition to the opportunity to suppress development and infection. We provide a basic overview of several strategies, including previously unpublished results, used by this laboratory to isolate stage-specific genes from Eimeria bovis. Additionally, we have included detailed discussions that summarize the associated advantages and disadvantages of each as applied to coccidia and potentially to other parasites in the phylum Apicomplexa. Given that the purification of sufficient quantities of high-quality RNA is vital, we have included detailed protocols for the isolation of RNA from various parasite stages. Also included is a detailed protocol to apply mRNA differential display to investigate stage specific developmental regulation. PMID- 9405200 TI - Techniques for isolation and characterization of apical organelles from Eimeria tenella sporozoites. AB - Apical organelles contain molecules that are of critical importance for the interaction of all apicomplexan parasites with their target host cells. Thus, there is considerable interest in characterizing and understanding the function of molecules that reside in these organelles. Large numbers of surface-sterilized oocysts of Eimeria tenella, an apicomplexan coccidian of the chicken, can be routinely obtained from the animal host, and invasive sporozoites, which contain abundant apical organelles, can be rapidly prepared from these oocysts in the laboratory. Thus, E. tenella is proving to be an amenable parasite for subcellular fractionation techniques that allow the direct isolation and characterization of apical organelles. In this paper, a series of protocols is described for the large-scale culture of E. tenella parasites, the preparation of invasive sporozoites, the isolation of apical organelles, and the use of in vitro culture for localization and functional studies. PMID- 9405201 TI - Immunochemical methods for identification of Babesia bovis antigens expressed on the erythrocyte surface. AB - Intraerythrocytic parasites, such as Babesia bovis, modify the erythrocyte plasma membrane structurally, antigenically, and functionally. For such parasites the infected erythrocyte surface also is thought to be a primary site for interaction with the host immune system. These properties demand characterization of the various alterations to understand the overall host-parasite interaction, immunity to disease or infection, and bases for parasite persistence. A paucity of adequate methods exists for characterization of parasite-derived components of the parasitized erythrocyte surface. To facilitate such studies we developed or modified several techniques to detect, identify, and localize parasite-induced alterations on the B. bovis-infected erythrocyte surface. These methods, which we present here, should be adaptable to a variety of intraerythrocytic parasite-host combinations. PMID- 9405203 TI - Penetration of Listeria monocytogenes in mice infected by the oral route. AB - In this study, it is suggested that the Peyer's patches are the most important point of entry of Listeria monocytogenes in the host after subclinical infection by the oral route. Microbiological, histopathological and ultrastructural evidence of infection was obtained in mice inoculated with a sublethal dose of 10(9) cfu. No mortality was observed. L. monocytogenes was isolated from the mesenteric lymph nodes from 6 hours post infection (hpi) through day 7 p.i. and from the liver and spleen from 24 h p.i. until days 5 and 7 p.i. respectively. Lesions were mainly restricted to the dome area of Peyer's patches and consisted of a purulent to pyogranulomatous inflammatory reaction. Scarce and minor lesions were also observed in the mesenteric lymph nodes and liver. L. monocytogenes was detected by immunohistochemistry in the Peyer's patches from 12 h p. i. to day 6 p.i. Ultrastructural study of Peyer's patches showed that the majority of Listeria cells were free within the cytoplasm of neutrophils and macrophages, not surrounded by a phagosomal membrane, and some of them were dividing. PMID- 9405202 TI - Yeast as a model system to study drugs effective against apicomplexan proteins. AB - Biochemical and genetic analyses are required to identify potential drug targets in apicomplexan parasites, but these studies have proved difficult in most parasite systems. We have developed methods based on expression of parasite proteins in the budding yeast, Saccharomyces cerevisiae, to rapidly screen drugs directed against particular parasite targets, to study the structure and function of these target molecules, and to identify mutations in the parasite genes that alter enzyme specificity or drug sensitivity. In this paper we outline the parameters that need to be considered to design yeast strains that function efficiently to assay function of parasite proteins. Basic protocols and methods are included. We detail some problems that might be encountered in the engineering of these yeast strains and suggest possible solutions. PMID- 9405204 TI - Streptococcal pyrogenic exotoxin A (SPE A) superantigen induced production of hematopoietic cytokines, IL-12 and IL-13 by human peripheral blood mononuclear cells. AB - A quantitative and kinetic study of the release of the hematopoietic cytokines IL 3, IL-5 and GM-CSF, the immunoregulatory cytokine IL-12 heterodimer (and its p40 subunit) and IL-13 by human peripheral blood mononuclear cells (PBMC) stimulated in vitro with the superantigen streptococcal pyrogenic (erythrogenic) exotoxin A (SPE A) from Streptococcus pyogenes is reported. PBMC were stimulated in parallel with heat-killed group A streptococcal cells, E. coli lipopolysaccharide (LPS) and with concanavalin A (Con A) in certain experiments for comparative purposes. The cytokines were assayed in the supernatant fluids by ELISA. IL-13 expression was also determined by a quantitative competitive PCR. IL-3, IL-5, GM-CSF, IL-12 p40, IL-12 heterodimer and IL-13 expression was induced by SPE A in a time- and dose-dependent manner in rather substantial amounts except the IL-12 heterodimer, which was released in small quantities. In contrast to SPE A, IL-3, IL-5 and IL 13 were not or poorly elicited by streptococcal cells or LPS whereas these two stimulants induced relatively high amounts of GM-CSF. Interestingly, both IL-12 p40 and IL-12 heterodimer were released in much higher amounts by streptococcal cells. Con A induced IL-3, IL-5, GM-CSF and IL-13 production in amounts comparable to those elicited by SPE A. The possible pathophysiological relevance of the elicitation by SPE A and streptococcal cells of these cytokines is discussed. PMID- 9405205 TI - Characterization of the Pasteurella haemolytica transferrin receptor genes and the recombinant receptor proteins. AB - The tbpA and tbpB genes encoding the transferrin receptor proteins, TbpA and TbpB, from Pasteurella haemolytica A1 were cloned, sequenced and expressed in Escherichia coli. The genes were organized in a putative operon arrangement of tbpB- tbpA. The tbpB gene was preceded by putative promoter and regulatory sequences, and followed by a 96 base pair intergenic sequence in which no promoter regions were found, suggesting that the two genes are coordinately transcribed. The deduced amino acid sequences of the TbpA and TbpB proteins had regions of homology with the corresponding Neisseria meningitidis, N. gonorrhoeae, Haemophilus influenzae and Actinobacillus pleuropneumoniae Tbp and Lbp proteins. The intact tbpB gene was expressed in a T7 expression system and the resulting recombinant TbpB protein retained the functional bovine transferrin binding characteristics. The availability of the recombinant TbpB enabled us to demonstrate its specificity for ruminant transferrins, its ability to bind both the C-and N-terminal lobes of bovine transferrin and its preference for the iron loaded form of this protein. Several attempts at expressing the cloned tbpA gene were unsuccessful, suggesting that the product of the gene may be toxic to E. coli. PMID- 9405206 TI - Local and systemic immune responses in humans against Helicobacter pylori antigens from homologous and heterologous strains. AB - The capacity of Helicobacter pylori to induce strain specific immune responses was studied in adult Swedish volunteers. Sera and gastric aspirates from 11 H. pylori-infected subjects were tested for specific antibody levels against, respectively, lipopolysaccharides (LPS) and total membrane preparations (MPs) prepared from the study subjects' own strains, as well as with corresponding antigens from two reference H. pylori strains or heterologous strains collected from other subjects within the study. It was found that sera from five of the 11 subjects had significantly higher IgA antibody titres against LPS from the homologous strain than against LPS from either of the reference strains and in five cases sera reacted with higher IgG titres against the homologous LPS than with LPS preparations from the reference or heterologous patient strains. Analyses of specific titres against MPs revealed that six sera had higher IgA titres and four sera had higher IgG titres against MPs prepared from the subjects' own strains than against MPs from either of the two reference strains. Determination of specific antibodies in gastric aspirates revealed significantly higher IgA titres against LPS from the homologous H. pylori isolate than against LPS from the two reference strains in five cases, and six aspirates reacted in higher IgA titre with the homologous H. pylori MPs. Results from immunoblotting analyses of sera support induction of strain specific immune responses against H. pylori LPS. By means of specific monoclonal antibodies against H. pylori LPS, antigenic heterogeneity between the different LPS preparations tested was confirmed. PMID- 9405207 TI - Loss of biological activity due to Glu-->Arg mutation at residue 11 of the B subunit of cholera toxin. AB - Since it has been reported that a single amino acid mutation of Gly-->Arg in the CAGYC region of the beta chain of human thyroid stimulating hormone (hTSH) was responsible for congenital isolated TSH deficiency, and that the same amino acid substitution in this site of hTSH and human chorionic gonadotropin (hCG) introduced by site-directed mutagenesis resulted in loss of activity, the authors studied the role of glutamic acid at position 11 (Glu-11) from the N-terminus of the B subunit of cholera toxin (CT), which corresponds to the glycine in the CAGYC region of the beta chain of hTSH and hCG. A mutant CT constructed by site directed mutagenesis in which Glu-11 was replaced by Arg (CT-E11R) did not induce either morphological changes or accumulation of cytosolic cyclic AMP in Chinese hamster ovary cells, although it formed the holotoxin AB5, retained the ability to bind to GM1-ganglioside and showed ADP-ribosyltransferase activity. Weak assembly of the B subunits in mutant CT-E11R demonstrated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under non-heating conditions might explain the loss of biological activity. PMID- 9405208 TI - Characterization of the transcriptional initiation regions of genes for the major secreted protein antigens 85C and MPB51 of Mycobacterium bovis BCG. AB - The component of mycobacterial 85 complex (85A, 85B, and 85C) and MPB51 are very important from immunological, biochemical, and antimycobacterial points of view. In this study, the transcriptional properties of genes encoding three components of 85 complex and MPB51 from BCG were analysed. The authors' analyses revealed that genes for 85A and MPB51 were transcribed as a single unit despite the one operon-like structure and these four genes were probably under a different regulatory control. These findings may help to understand the immunological and physiological roles of these antigens. PMID- 9405209 TI - Salmonella typhimurium aroB mutants are attentuated in BALB/c mice. AB - The aroB gene of Salmonella typhimurium, encoding dehydroquinate synthase, has been cloned into pUC19 and the DNA sequence determined. The aroB gene was isolated from a cosmid gene bank by complementation of an Escherichia coli aroB mutant and screening by Southern blot analysis. The nucleotide sequence of the S. typhimurium aroB gene revealed the presence of an open reading frame, encoding a protein of 362 amino acids with a calculated molecular mass of 38696 Daltons. The amino acid sequence of S. typhimurium dehydroquinate synthase is nearly identical to the E. coli homologue and shows high homology with other aroB gene products from other organisms. Subsequently, a stable insertional mutation in aroB was introduced into the wild-type S. typhimurium C5 strain. This mutant was auxotrophic for aromatic compounds. Infection of BALB/c mice with this mutant demonstrated attenuation comparable to other S. typhimurium mutants unable to biosynthesize aromatic amino acids. PMID- 9405210 TI - Internal rotation of mutually interacting methyl groups: A 13C NMR study AB - The overall and intramolecular rotational diffusion behavior of 1,3, 7,10 tetramethylbenzo[c]cinnoline was determined from longitudinal 13C NMR relaxation and 1H-13C NOE measurements in dilute chloroform solution. The four methyl groups in this compound represent three different situations of sterical hindrance. One pair of methyl groups is in close mutual sterical contact, forming the ends of an open six-membered ring. Assuming completely anisotropic overall molecular tumbling combined with a 120 degrees jump model for the internal methyl rotations the jump rates of methyl groups were evaluated and compared to earlier results on different sterically hindered compounds, in particular with respect to a potential cogwheel-like intermethyl interaction. To characterize intermethyl interactions in different sterical situations, a new gauge-the "methyl interaction volume"-is introduced. Implications for correlated rotational diffusion of methyl groups are discussed. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9405211 TI - Quadrupolar relaxation of spin 3 in the intermediate omega0tauc regime AB - The equations for the quadrupolar relaxation of spin 3 were derived for the Redfield limit where the molecular reorientation rate is much faster than the size of the quadrupolar interaction. In the extreme narrowing regime (omega0tauc << 1), the results converge to the analytical expressions for the relaxation rates available in the literature. For slower motions, both longitudinal (spin lattice) and transverse (spin-spin) relaxations are described by a superposition of three exponentials, where both the rates themselves and their relative weights are functions of omega0tauc. Numerical calculations of the relevant relaxation parameters in the intermediate omega0tauc regime are presented. Spin-lattice relaxation is described to very good approximation by a single exponential for all values of omega0tauc, with the weight of the dominant decay mode exceeding 0. 97 for the entire range. The predictions of these simulations were found to be in good agreement with experimentally measured relaxation rates of the 10B resonances in the sodium salt of Na2B12H12S, mercaptoundecahydro-closo dodecaborane (sodium borocaptate or BSH) dissolved in glycerol, determined at omega0 = 53. 73 MHz, between temperatures of 268 and 323 K. The fit to the experimental results yielded a value of 1.25 MHz for the average 10B quadrupolar coupling constant in this molecule. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9405212 TI - A method for simulation of NOESY, ROESY, and off-resonance ROESY spectra AB - A formalism is proposed for simulation of NOESY, ROESY, and, more specifically, off-resonance ROESY nuclear magnetic resonance spectra. The off-resonance ROESY experiment has several advantages compared to standard NOESY and ROESY experiments. A simplified formalism which allows rapid computer simulation of the development of magnetization, including relaxation, in the presence of an RF field is of general use, in particular in the implementation and interpretation of off-resonance ROESY experiments. The relevant matrix equations can be derived either from the classical Bloch and Solomon equations or from the quantum mechanical homogeneous master equation in the basis of the Cartesian product operators. Examples of simulated spectra and behavior of magnetization during pulse sequences are shown. In addition, we present the full quantum mechanical theory for a two-spin system derived from the homogeneous master equation. The proposed formalism, here applied to off-resonance ROESY, has many potential applications, e.g., in the development and analysis of ROESY and TOCSY mixing sequences, selective pulses, and decoupling in which the complete spin dynamics, including relaxation, is taken into account. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9405213 TI - Coherence sidebands in adiabatic decoupling AB - RF pulse sequences applied to IS spin systems may produce substantial transverse antiphase S magnetization coupled to antiphase I magnetization, just prior to detection of the S signal, for samples containing a range of J coupling constants, or when pulse sequence delays are misset from ideal values. This magnetization is generally considered to be unobservable. Adiabatic decoupling on the I spins during signal detection efficiently converts this magnetization to observable S signal in the form of sidebands which we dub "coherence sidebands." Three single-transient pulsed-field-gradient methods are described for eliminating these unwanted sidebands. The techniques are applicable to 1H detected 13C-decoupled experiments on spectrometers operating at a 1H frequency of up to 2 GHz. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9405214 TI - Improved method for accurate and efficient quantification of MRS data with use of prior knowledge AB - We introduce AMARES (advanced method for accurate, robust, and efficient spectral fitting), an improved method for accurately and efficiently estimating the parameters of noisy magnetic resonance spectroscopy (MRS) signals in the time domain. As a reference time domain method we take VARPRO. VARPRO uses a simple Levenberg-Marquardt algorithm to minimize the variable projection functional. This variable projection functional is derived from a general functional, which minimizes the sum of squared differences between the data and the model function. AMARES minimizes the general functional which improves the robustness of MRS data quantification. The newly developed method uses a version of NL2SOL, a sophisticated nonlinear least-squares algorithm, to minimize the general functional. In addition, AMARES uses a singlet approach for imposition of prior knowledge instead of the multiplet approach of VARPRO because this greatly extends the possibilities of the kind of prior knowledge that can be invoked. Other new features of AMARES are the possibility of fitting echo signals, choosing a Lorentzian as well as a Gaussian lineshape for each peak, and imposing lower and upper bounds on the parameters. Simulations, as well as in vivo experiments, confirm the better performance of AMARES compared to VARPRO in terms of accuracy, robustness, and flexibility. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9405215 TI - 2H double-quantum NMR spectroscopy for the study of molecular motion in solids AB - An experiment which separates 2H spinning sideband patterns according to the 2H double-quantum chemical shift in a manner suitable for motional studies is proposed. The technique is successfully applied to 4,4'-azoxydianisole at 323 K to separately analyze the motion of the aromatic and methyl deuterons. Comments are made on the formation of a nematic mesophase in this material at 390 K in the light of the results of this NMR study. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9405216 TI - 1D and 2D MAS NMR spectra of a dipolar-coupled homonuclear spin-(1/2) pair AB - An analytical approach based on the average Hamiltonian theory is proposed for efficient calculation of the MAS NMR spectra of a dipolar-coupled homonuclear spin-(1/2) pair. For this purpose a superoperator formalism is developed which allows one to describe the spectra over a broad span of sample spinning rates, including the exact rotational resonances. This formalism can also be applied to the description of 2D polarization exchange spectra, which in many cases turns out to be useful for measuring the coupling strength. The experimental MAS NMR and the 2D spectra of doubly 13C-labeled zinc acetate were found to be in good agreement with the calculated spectra. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9405217 TI - Three-dimensional maximum-quantum correlation HMQC NMR spectroscopy (3D MAXY HMQC) AB - The extension of two-dimensional maximum-quantum correlation spectroscopy (2D MAXY NMR), which can be used to simplify complex NMR spectra, to three dimensions (3D) is described. A new pulse sequence for 3D MAXY-HMQC is presented and exemplified using the steroid drug dexamethasone. The sensitivity and coherence transfer efficiency of the MAXY NMR approach has also been assessed in relation to other HMQC- and HSQC-based 3D methods. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9405218 TI - A simple matrix formalism for spin echo analysis of restricted diffusion under generalized gradient waveforms AB - A simple mathematical formalism is presented which allows closed form expressions for the echo attenuation, E(q), in spin echo diffusion experiments, for practically all gradient waveforms and for the case of restricted diffusion in enclosing pores, with or without wall relaxation. The method, which derives from the multiple propagator approach of A. Caprihan et al. (1996, J. Magn. Reson. A 118, 94), depends on the representation of the gradient waveform by a succession of sharp gradient impulses. It leads to E(q) being expressed as a product of matrix operators corresponding quite naturally to the successive sandwich of phase evolution and Brownian migration events. Simple expressions are given for the case of the finite width gradient pulse PGSE experiment, the CPMG pulse train used in frequency-domain modulated gradient spin echo NMR, and the case of a sinusoidal waveform. The finite width gradient pulse PGSE and CPMG pulse trains are evaluated for the case of restricted diffusion between parallel reflecting planes. The former results agree precisely with published computer simulations while the latter calculation provides useful insight regarding the spectral density approach to impeded Brownian motion. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9405219 TI - Coupling amplification in 2D MAS NMR and its application to torsion angle determination in peptides. AB - A technique for amplifying the apparent magnitudes of 13C-1H and 15N-1H dipolar interactions in magic-angle spinning experiments is described. By inserting rotor synchronized 180 degrees pulses in the evolution period of a 2D dipolar-chemical shift experiment, heteronuclear dipolar couplings are doubled or quadrupled relative to the spinning speed. The increased number of dipolar sidebands is desirable for retaining structural information in the indirectly detected dipolar dimension while resolving inequivalent sites in the isotropic chemical shift dimension at relatively high spinning speeds. This coupling amplification method is incorporated into an experiment that determines the peptide torsion angle phi through the relative orientation of the Calpha-Halpha and N-HN bonds. It is shown both experimentally and theoretically that the angular resolution of the measurement is enhanced significantly by the selective doubling of the N-HN coupling. PMID- 9405220 TI - ROESY with water flip back for high-field NMR of biomolecules. AB - We report a version of the ROESY experiment in which saturation of the water magnetization is avoided without compromising suppression of the water signal during acquisition. Field gradient and selective RF pulses are used to maintain precise control of the water magnetization throughout the experiment and avoid signal losses due to radiation damping and molecular diffusion effects. The pulse sequence includes a delay for intentional radiation damping prior to mixing period. The optimal length of this delay is field and sample dependent, but easily determined from the apparent linewidth of the water signal. NOESY and TOCSY variants of the same experiment are presented which make use of identical manipulations of the water magnetization. The three pulse sequences constitute a suite for which little parameter adjustment is required once one of the experiments has been configured. PMID- 9405221 TI - Discrimination between the different compartments in sciatic nerve by 2H double quantum-filtered NMR. AB - The 2H double-quantum-filtered (DQF) NMR spectrum of isolated rat sciatic nerve, equilibrated with deuterated saline, is composed of three quadrupolar-split water signals. On the basis of the time course of their shift by Co-EDTA2- and CoCl2, the signals with quadrupolar splittings of about 120, 470, and 9 Hz were assigned to water in the epineurium, endoneurium, and intra-axonal compartments, respectively. The signal of the bulk water, which experiences isotropic motion, was eliminated by the DQF pulse sequence. As the maximum intensities of the water signals in the three anisotropic compartments occur at different creation times, in the DQF pulse sequence, it is possible to resolve the signals and measure their properties, such as relaxation times, independently, without perturbing the system with shift reagents. PMID- 9405222 TI - Nuclear magnetic resonance imaging of solid rocket propellants at 14.1 T. AB - Proton NMR images of solid propellant materials, consisting of a polybutadiene binder material filled with 82% solid particles, have been obtained at a magnetic field strength of 14.1 T and at a resolution of 8.5 x 8.5 micron. The images are the first of elastomeric materials obtained at a proton frequency of 600 MHz and have the highest spatial resolution yet reported. The images display a high contrast and are rich in information content. They reveal the distribution of individual filler particles in the polymer matrix as well as a thin polymer film of about 10-30 micron which is found to surround some of the larger filler particles. PMID- 9405224 TI - Regulatory elements in the insulin-responsive glucose transporter (GLUT4) gene. AB - GLUT4, the insulin responsive-glucose transporter, mediates the rate limiting step of glucose metabolism in skeletal muscle and adipose tissue. GLUT4 expression is up-regulated by exercise training and thyroid hormone treatment and is down-regulated by fasting, streptozotocin-induced diabetes, obesity, high-fat diet, and denervation. Since overexpression of GLUT4 in insulin resistant db/db mice and high-fat diet-fed mice has been observed to dramatically improve glycemic control, increasing GLUT4 expression may be an effective strategy with which to alleviate insulin resistance. This review discusses recent findings on the regulation of the GLUT4 gene and on progress in the identification of regulatory elements in the promoter of the gene. PMID- 9405225 TI - Tetrahydrobiopterin modulates cyclooxygenase-2 expression in human mesangial cells. AB - Tetrahydrobiopterin (BH4) biosynthetic pathways are stimulated under inflammatory conditions. The newly synthesized BH4 serves as a cofactor for optimal activity of inducible nitric oxide synthase (NOS2). In human mesangial cells (HMC), BH4 is also a limiting factor for NOS2 expression. In this study we show that BH4 availability can also play a modulatory role in the expression of cyclooxygenase 2 (COX-2) in HMC. Supplementing HMC with the BH4 donor sepiapterin potentiated IL 1beta/TNF-alpha-induced COX-2 expression by approximately 2-fold. This effect was abolished by methotrexate. In contrast, the NOS inhibitor L-NAME and the soluble guanylate cyclase inhibitor ODQ did not block sepiapterin amplification of COX-2 expression. Moreover, sepiapterin was found to modulate the tyrosine phosphorylation of several cellular substrates, an early event which occurred well before the induction of NOS2 could be evidenced. These findings suggest a role for BH4 in the modulation of mesangial cell responses to pro-inflammatory stimuli. PMID- 9405226 TI - Haloenol lactone: a new synergist of chemotherapy in vitro. AB - Over-expression of glutathione S-transferases (GST) has been found to play a significant role in multiple drug resistance in cancer chemotherapy. To combat GST-mediated drug resistance, GST inhibitors are being studied as potential synergists for effective cancer chemotherapy. We have designed and synthesized a haloenol lactone derivative as a mechanism-based inactivator of GST-pi isozyme. In the current study, we examined the inhibitory effect of the haloenol lactone compound on GST of a human renal carcinoma cell line UOK130 and found that this compound shows time-dependent GST inhibition in these cancer cells. The enzyme activity lost upon incubation with the haloenol lactone could not be restored by extensive dialysis against buffer. Pretreatment of the cancer cells with 1.0 microM of haloenol lactone increased cytotoxicity induced by cisplatin in the UOK130 cell line. This report further supports the possibility of synergizing alkylating agents in cancer chemotherapy by use of selective GST inhibitors. PMID- 9405227 TI - Influence of molecular weight, protein core and charge of native heparin fractions on pulmonary artery smooth muscle cell proliferation. AB - Heparin macromolecules have been shown to inhibit cultured pulmonary artery smooth muscle cell proliferation in vitro and prevent hypoxic vascular remodeling in vivo. In an attempt to understand the structural determinants of heparin's antiproliferative properties, we have fractionated an antiproliferative preparation of commercial heparin into low and high molecular weight fractions. Then the high molecular weight heparin fraction was further fractionated on a DEAE-cellulose column by charge density eluting with 0 - 1 M NaCl linear gradient. The heparin protein peptides were both removed and isolated. These heparin fractions were assayed for antiproliferative effects on cultured bovine pulmonary artery smooth muscle cells. No appreciable differences were found among high and low molecular weight heparin fractions The core peptides showed no antiproliferative activity. However, higher charge density fraction was less antiproliferative. PMID- 9405228 TI - Overexpression of members of the AP-1 transcriptional factor family from an early stage of renal carcinogenesis and inhibition of cell growth by AP-1 gene antisense oligonucleotides in the Tsc2 gene mutant (Eker) rat model. AB - We previously isolated subtracted cDNA clones for genes having increased expression in Tsc2 gene mutant (Eker) rat renal carcinomas (RCs). Among them, fra 1 encoding a transcriptional factor activator protein 1 (AP-1) was identified. We have therefore investigated whether other members of the AP-1 transcription factor family might also be involved in renal carcinogenesis in the Eker rat model. In the present study, overexpression of fra-1, fra-2, c-jun, junB, and junD mRNAs was demonstrated in RCs by Northern blot analysis. Interestingly, AP-1 proteins were highly expressed even in the earliest preneoplastic lesions (e.g., phenotypically altered tubules) as suggested by immunohistochemistry. Moreover, 12-O-tetradecanoylphorbol-13-acetate-responsive element (TRE)-binding activity of AP-1 proteins was observed in RC cell extracts by electrophoretic mobility shift assay. As a next step, we transfected antisense oligonucleotides targeting AP-1 genes into RC cells and demonstrated that their growth was strongly inhibited. Thus, the data suggest that overexpression of AP-1 genes might play a crucial role in renal carcinogenesis in the Eker rat model. PMID- 9405229 TI - The interaction of HIV-1 Tat(32-72) with its target RNA: a fluorescence and nuclear magnetic resonance study. AB - We performed intrinsic peptide fluorescence experiments to characterize the interaction between variants of the HIV-1 Tat(32-72) peptide BP1 and TAR RNA. Kd values for wild-type BP1 and cysteine-modified BP1 were found to be in the range of 60 to 70 nM for both peptides, indicating that free sulfhydryl groups of the cysteines within the peptide are not required for high affinity TAR binding. Thus, the mutant peptide BP1 (C34S, C37W) (BP1SW) was used to further investigate peptide RNA interaction by fluorescence studies. Titration of BP1SW with TAR resulted in a dissociation constant (Kd = 9 nM) nearly an order of magnitude lower than that of the wild-type peptide. The change of the BP1SW fluorescence intensity on TAR binding was used to investigate the kinetics of this interaction by stopped-flow experiments. The results can be explained in terms of a two-step binding model, with a rapid diffusion-limited initial formation of a tight, but unspecific peptide RNA complex, followed by a relatively slow structural rearrangement step (k approximately 60 s-1) in order to form the specific BP1SW TAR complex. Comparison of heteronuclear two-dimensional NMR spectra of BP1SW and BP1SW bound to TAR shows that only resonances from amino acid residues of the core and basic sequence regions are shifted on TAR binding. PMID- 9405230 TI - Peroxynitrite protects RAW 264.7 macrophage from Lipopolysaccharide/Interferon gamma-induced cell death. AB - Peroxynitrite, formed by the interaction of superoxide with nitric oxide, has previously been implicated mostly as a cytotoxic agent. In contrast, its physiological and, possibly, beneficial effects are largely unknown. We have previously shown [Journal of Biological Chemistry, 1997, 272, 7253] that RAW 264.7 macrophages can be selected to be resistant toward lipopolysaccharide (LPS)/interferon-gamma (IFN-gamma)-induced cytotoxicity. Resistant cells produced comparable amount of nitric oxide, but showed increased formation of superoxide, which might lead to increased production of peroxynitrite. We utilized this well characterized cell model to seek evidence that peroxynitrite might cause protection of RAW cells from cytokine toxicity. Exogenous peroxynitrite (30-50 microM), applied to RAW cells before cytokine stimulation, dramatically reduced LPS/IFN-gamma toxicity. Measurement of cell viability after overnight incubation with a mixture of LPS (10 microg/ml) and IFN-gamma (100 U/ml), showed that pretreatment with 40 microM peroxynitrite completely reverted LPS/IFN-gamma cytotoxicity. Differently, pretreatment of RAW cells with peroxynitrite (10-60 microM) did not prevent cytotoxicity induced by the nitric oxide-donors S-Nitroso L-glutathione (0.2-1 mM), or spermine NONOate (0.2-2 mM), and by Actimomycin D (0.5-1 microg/ml), suggesting that the protective effect is specific for the LPS/IFN-gamma pathway. These results were confirmed through extensive controlled studies aimed to optimize cell exposure to peroxynitrite, and showed that peroxynitrite protects macrophages from cytokine-induced cytotoxicity. PMID- 9405231 TI - Evaluation of anti-hepatitis B virus (HBV) drugs using the HBV transgenic mouse: application of the semiquantitative polymerase chain reaction (PCR) for serum HBV DNA to monitor the drug efficacy. AB - For evaluation of anti-hepatitis B virus (HBV) drugs, we have employed the HBV transgenic mouse in which virion-like particles can be assayed in the serum. Bispivaloyloxymethyl-9-(2-phosphonylmethoxyethyl)-adenine [bis (POM) PMEA] 100 mg/kg/day, 2',3'-dideoxy-3'-thiacytidine [(+-)-BCH189] 200 mg/kg/day and a placebo were orally administered to mice twice a day for 14 days. Anti-viral effects were monitored by checking the levels of serum HBV DNA by the semiquantitative polymerase chain reaction, HBsAg and HBeAg by enzyme immunoassay, and replicative intermediates in the liver by Southern blotting. As expected, decrease from the 10(0.5) to 10(3) copies of HBV DNA per microl of sera detected before the treatment to the undetectable level was evident for all five animals treated with bis(POM) PMEA 100 mg/kg/day. However (+-)-BCH189 200 mg/kg/day, which is known to act as the inhibitor of reverse transcriptase for HBV or HIV in vivo and in vitro, did not suppress HBV DNA levels in the transgenic mouse. Thus, we were able to detect the effects of anti-HBV drugs semi quantitatively, and confirm differences in drug efficacy. PMID- 9405232 TI - Mechanism of depletion of liver glycogen in cancer cachexia. AB - Mice transplanted with a cachexia-inducing colonic adenocarcinoma (MAC16) show a progressive decrease in liver glycogen in direct proportion to the loss of body weight. Such tumours elaborate a lipid mobilizing factor (LMF), which produces a dose-dependent stimulation, not only of adipocyte adenylate cyclase, but also of hepatocyte adenylate cyclase in a GTP-dependent manner. These results suggest that LMF has the capacity to initiate hepatic glycogenolysis through an increase in cyclic AMP. PMID- 9405233 TI - Molecular cloning and expression of a novel human aquaporin from adipose tissue with glycerol permeability. AB - In a systematic analysis of genes expressed in human adipose tissue, we detected a novel gene that is expressed uniquely in adipose tissue. The sequence showed that it encodes a 342-amino-acid protein containing six putative transmembrane domains, and is a new member of the aquaporin family of water-selective membrane channels. We named this gene aquaporin 9. It features a cyclic-AMP protein kinase phosphorylation consensus site in the NH3-terminal domain. Expression of the cRNA in Xenopus oocytes yielded a 7-fold increase in osmotic water permeability blocked by 0.3 mM HgCl2, and also facilitated the uptake of glycerol. Northern blot analysis demonstrated that the mRNA is abundant in adipose tissue, but not in other tissues. Thus, this gene product may participate in glycerol transport in adipocytes. PMID- 9405234 TI - A bidirectional promoter connects the p14.5 gene to the gene for RNase P and RNase MRP protein subunit hPOP1. AB - We have identified the functional promoter of the translational inhibitor p14.5, the human homologue to a rat perchloric acid-soluble protein (PSP), a mouse heat responsive protein (Hrp12) and a goat tumor antigen (UK114). Sequence analysis revealed a GC-rich promoter with several consensus sequences for transcription factors, but no TATA- and CAAT-box. To confirm promoter activity, DNA fragments of the p14.5 5'-flanking region were ligated in front of the luciferase gene and were transfected into HeLa and HepG2 cells. A minimal promoter between nt -104 and nt +88 relative to the transcription start site was responsible for basal activity. Furthermore, we observed a head-to-head orientation of p14.5 to the gene for the protein subunit of RNase P and MRP ribonucleoproteins (hPOP1). Luciferase assays with fragments of the hPOP1 5'-flanking region revealed a minimal promoter between nt -20 and nt +98 relative to the start of transcription. These data indicate that the 102 bp region between p14.5 and hPOP1 can act as a bidirectional promoter. The p14.5-hPOP1-cluster was mapped to chromosome 8q22 using in situ hybridization technique. PMID- 9405235 TI - The rat serotonin transporter: identification of cysteine residues important for substrate transport. AB - Reduction and alkylation of disulfide bonds are known to affect substrate translocation by and antidepressant binding to the serotonin transporter (SERT). To identify functionally relevant cysteine residues, we substituted 16 cysteins of the rat SERT by alanine or serine residues and analyzed the transport and binding properties of the respective mutant transporters after heterologous expression in a mammalian cell line. Replacement of cysteine 209 by serine resulted in a marked reduction of the maximal transport rate, loss of positive cooperativity, and insensitivity to treatment with disulfide reducing agents, indicating that cysteine 209 participates in a structurally important disulfide bridge. Replacement of cysteine residues 147, 200, 369, and 540 caused a complete loss of both substrate transport and antidepressant binding, a result that is likely to reflect impaired processing and/or cell surface expression of the mutated polypeptides. PMID- 9405236 TI - Serine base exchange enzyme activity is modulated by sphingosine and other amphiphilic compounds: possible role of positive charge in increasing the synthesis of phosphatidylserine. AB - It has been found that sphingosine and sphingosylphosphorylcholine (amphiphilic cations) have a stimulatory, and cholesterol 3-sulfate (an amphiphilic anion), an inhibitory, effect on [14C]serine incorporation into phosphatidylserine in glioma C6 and rat liver microsomes. In glioma intact cells sphingosine stimulates phosphatidylserine synthesis in a process independent of protein kinase C, but suppressed by thapsigargin. We suggest that the stimulation of the enzyme occurs by the interaction of amphiphilic cations with the membrane cosubstrate phospholipids, leading to a charge redistribution on their phosphate groups, and hence facilitating the enzyme action. A new hypothesis concerning the mechanism of the serine base exchange reaction is discussed. PMID- 9405237 TI - Evidence for CYP2D6 expression in human lung. AB - The cytochrome P450 CYP2D6 gene (CYP2D6) expression was examined in samples from human bronchial mucosa and lung parenchyma using reverse transcription-polymerase chain reaction (RT-PCR) and immunochemistry. Except specimen from a patient previously genotyped as homozygous for a complete deletion of the gene, all tissue samples were positive. When compared to that in the liver, the mean level of CYP2D6 mRNA was 3-fold lower in bronchial mucosa and 6-fold lower in lung parenchyma. To our knowledge, these data demonstrate for the first time the presence of CYP2D6 protein in human lung. They also indicate that the gene is nonuniformly distributed within this organ. The possibility that CYP2D6 has a role in lung carcinogenesis by locally activating inhaled chemicals should therefore be considered. PMID- 9405239 TI - A novel DNA virus (TTV) associated with elevated transaminase levels in posttransfusion hepatitis of unknown etiology. AB - By means of representational difference analysis, a viral clone (N22) of 500 nucleotides was isolated from serum of a patient (TT) with posttransfusion hepatitis of unknown etiology. The N22 clone showed a poor homology to any reported sequences. Oligonucleotide primers were deduced from the N22 sequence for detecting it by polymerase chain reaction. N22 sequence in serum banded at a sucrose density of 1.26 g/cm3, indicating its association with a viral particle which was designated TT virus (TTV). Since nucleic acids of TTV were sensitive to DNase I, it would be a DNA virus. TTV DNA was detected in sera from three of the five patients with posttransfusion non-A to G hepatitis, including the index case (TT). TTV DNA titers closely correlated with aminotransferase levels in the three patients. These results indicate that TTV would be a novel DNA virus with a possible capacity to induce posttransfusion non-A to G hepatitis. PMID- 9405240 TI - Involvement of hepatocyte growth factor in formation of bronchoalveolar structures in embryonic rat lung in primary culture. AB - To clarify the role of hepatocyte growth factor (HGF) in embryonic lung development, organoids from fetal rat lung were cultured in collagen gels with or without HGF antisense oligonucleotides. Cyst-like structures formed within 24 h in organoids isolated from fetuses after 14 days' gestation, but this was abolished by the oligonucleotide addition, apparently by interference with the endogenous expression of HGF. Electron microscopy revealed two types of structure: an alveolar type characterized by osmiophilic lamellar bodies in the cytoplasm and lumen, and a bronchial type consisting of epithelial cells bearing microvilli on their apical surfaces. HGF mRNA was detectable from day 14 in fetal lung by RT-PCR. Our results suggest that HGF plays, coordinately with its expression, a crucial role in the morphogenesis of both alveolar and bronchial epithelia in the rat fetal lung. PMID- 9405238 TI - Transforming growth factor-beta1 coregulates mRNA expression of aryl hydrocarbon receptor and cell-cycle-regulating genes in human cancer cell lines. AB - Transforming growth factor (TGF)-beta1 down-regulates mRNA expression of the aryl hydrocarbon receptor (AhR) and of AhR-inducible genes in A549 cells. Here, we describe a dose-dependent inhibition of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-induced cytochrome P450 (CYP) 1A1, CYP1B1 and NADPH-quinone-oxidoreductase (NMO-1) mRNA expression as well as TCDD-induced 7-ethoxyresorufin-O-deethylase (EROD) activity in MDA-MB 231 cells. The AhR mRNA expression was not affected by TGF-beta1. TGF-beta-responsiveness was investigated by examining the effect on the expression of responsive genes. While TGF-beta1 up-regulates mRNA expression of TGF-beta1 and TIEG (TGF-beta-inducible early gene) as well as luciferase activity of a responsive reporter plasmid in both cell lines, a down-regulation of c-myc and cyclin A mRNA expression was only found in A549 cells. Furthermore, TGF-beta1 inhibits only cell proliferation of A549 but not of MDA-MB 231 cells. The results show a coregulation of mRNA expression of AhR and cell-cycle regulating genes, and further indicate that the AhR may be involved in regulation of cell proliferation. PMID- 9405241 TI - The quinoline-based drug, N-[4-[1-hydroxy-2-(dibutylamino)ethyl] quinolin-8-yl]-4 azidosalicylamide, photoaffinity labels the multidrug resistance protein (MRP) at a biologically relevant site. AB - MRP is a member of the ABC trafficking proteins thought to mediate the transport of glutathione S-conjugates and amphiphilic natural products. However, unlike P glycoprotein, the biochemical mechanism by which MRP mediates the resistance to cytotoxic drugs is not clear. In this report, we describe the interactions of a quinoline-based drug, N-{4-[1-hydroxy-2-(dibutylamino)ethyl] quinolin-8-yl}-4 azidosalicylamide (IAAQ), with MRP. Our results demonstrate the ability of IAAQ to photoaffinity label a 190 kDa protein in resistant Small Cell Lung Cancer cells (H69/AR) but not in the parental H69 cells. The photoaffinity labeling of the 190 kDa protein with IAAQ was both saturable and specific. The identity of the 190 kDa protein, as MRP, was confirmed by immunoprecipitation with the monoclonal antibody, QCRL-1. Furthermore, a molar excess of LTC4, MK 571 or vinblastine inhibited the photoaffinity labeling of MRP with IAAQ in intact cells and plasma membranes. Cell growth and drug transport studies showed H69/AR cells to be less sensitive to and to accumulate less IAAQ than the parental H69 cells. In addition, MK 571 and doxorubicin increased the sensitivity to and the accumulation of IAAQ in H69/AR cells. Together, the results of this study show for the first time the direct binding of unaltered cytotoxic drug to MRP. Moreover, given the structural similarities between IAAQ and MK 571, we suggest that MK 571 modulates MRP-mediated resistance by direct binding to MRP. PMID- 9405242 TI - Use of EBV-based Vector/HVJ-liposome complex vector for targeted gene therapy of EBV-associated neoplasms. AB - Targeted suicide gene therapy for Epstein-Barr virus (EBV)-associated neoplasms was attempted by using EBV-based plasmid vectors coupled with hemagglutinating virus of Japan (HVJ)-liposome in vitro. Expression of EBV nuclear antigen (EBNA)1 is a common feature of the neoplasms associated with EBV. When various leukemic cell lines were transduced with a vector carrying a marker gene and EBV replication origin of plasmid (oriP), the marker gene product was exclusively detected in cells expressing EBNA1. Transduction of herpes simplex virus (HSV)-1 thymidine kinase (Tk) gene resulted in a marked reduction in viable cell number by ganciclovir (GCV) specifically in EBNA1 positive cells. The results demonstrate that this virus-free system may be applicable to gene therapy of EBV associated neoplasms. PMID- 9405243 TI - Evidence of his61 imidazolate bridge rupture in reduced crystalline Cu,Zn superoxide dismutase. AB - X-ray absorption spectroscopy has been carried out on the copper K edge in oxidized and reduced bovine Cu,Zn SOD in solution and in crystalline state. The results indicate that the copper coordination geometry is unaffected by the solution or by the crystalline state of the protein, in both oxidation states. Moreover the two oxidation states of the active copper ion are reflected under, all the experimental conditions, by distinct coordination spheres around the catalytic metal, which is four-coordinated and three-coordinated in the Cu(II) and in the Cu(I) enzyme, respectively. PMID- 9405244 TI - Control of expression of PLCbeta1 by LAC repressor system: relationship between nuclear PLCbeta1 overexpression and growth factor stimulation. AB - Swiss 3T3 cells have a nuclear phosphoinositide signalling system which is under the control of insulin-like growth factor I (IGF-I) and acts separately from that at the plasma membrane. By using the Lac repressor system we were able both to obtain the inducible overexpression of phospholipase C beta1 (PLC beta1) and to determine its subcellular localisation and partitioning. Moreover, by comparing the level of expression at the nucleus and the percentage of cells actively incorporating bromodeoxyuridine (BrdU) in S phase it has strengthened the issue of the importance of this PLC in the onset of DNA synthesis mediated by IGF-I. In addition, this system appears to be a very powerful tool for further analysis of the downstream events following the activation of nuclear PLC beta1. PMID- 9405245 TI - Evidence for an asymmetrical uptake of L-carnitine in the blood-brain barrier in vitro. AB - The transport of L-carnitine (4-N-trimethylammonium-3-hydroxybutyric acid) was studied with a primary culture of porcine brain capillary endothelial cells (BCEC) as an in vitro model of the blood-brain barrier. The measurements with suspended cells and cell monolayers allowed to distinguish a polarized transport phenomena. The part of the BCEC cells exposed to the medium (apical membrane) accumulated carnitine by a sodium-independent, saturable (Km=28 microM) system, with k=0.018 min-1. Exposure of the basolateral part revealed a presence of a facilitated diffusion process. Carnitine uptake through the saturable system was inhibited by butyrobetaine. Acylcarnitines and choline have no effect on the carnitine accumulation in suspended cells, a process diminished by phenylalanine, leucine, and L system inhibitor. This points to the possibility that carnitine enters through the basolateral membrane using amino acid transporting systems. A different, novel system is postulated to operate in the apical part of the plasma membrane of BCEC. PMID- 9405246 TI - Protein half-lives of two subunits of an NMDA receptor-like complex, the 71-kDa glutamate-binding and the 80-kDa CPP-binding protein. AB - We determined the half-lives for two subunits of a complex that functions as a glutamate and N-methyl-D-aspartate (NMDA) receptor-ion channel in synaptic membranes. These two proteins are a 71 kDa glutamate-binding protein (GBP) and an 80 kDa CPP-binding protein (CBP). Seven month-old Fischer 344 rats were injected with L-[14C] leucine. The radioactivity in the two proteins was determined in a crude synaptosomal membrane fraction obtained from the brains of rats sacrificed from 4 hours to 13 days after the injection. The previously reported data on time dependent appearance and loss of L-[14C] leucine radioactivity in the serum (Ferrington et al., 1997, Biochem. Biophys. Res. Commun. 237, 163-165) was used in the present study to estimate the half-lives of GBP and CBP. Theoretical curves best fit the experimental data obtained for the two proteins assuming apparent half-lives of 14 (+/- 2.4) and 18 (+/- 1.2) hours for CBP and GBP, respectively. PMID- 9405247 TI - The effects of eotaxin on the surface adhesion molecules of endothelial cells and on eosinophil adhesion to microvascular endothelial cells. AB - Eosinophil recruitment occurs in tissues as the result of allergic diseases. Human eotaxin is thought to be specific to eosinophils. In this study, we examined the effects of human eotaxin on the expression of adhesion molecules on nasal microvascular endothelial cells and on eosinophil adhesion to endothelial cells. Eotaxin upregulated the expression of ICAM-1 and VCAM-1 on human nasal mucosal microvascular endothelial cells (HMMEC), but not human umbilical vein endothelial cells (HUVEC). The eotaxin-induced eosinophil adhesion to HMMEC was increased at 10 ng/ml and significantly increased at the concentration of 100 ng/ml. On HUVEC, however, eotaxin did not induce increases of eosinophil adhesion. Anti-ICAM-1 and anti-VCAM-1 mAbs significantly decreased eotaxin induced eosinophil adhesion. These results suggest that eotaxin regulates eosinophil accumulation to the nasal mucosa through its effect on the adhesion molecules on microvascular endothelial cells. PMID- 9405248 TI - Histone deacetylase inhibitor activates the WAF1/Cip1 gene promoter through the Sp1 sites. AB - Treatment of cultured cells with trichostatin A (TSA), a specific histone deacetylase inhibitor, induces the histone hyperacetylation and modulates expression of some mammalian genes. We examined the effects of TSA on cell growth arrest, and its relation to expression of the WAF1/Cip1 gene, a potent inhibitor of cyclin-dependent kinases, in a p53-mutated human osteosarcoma cell line MG63. TSA at 500 ng/ml induced growth arrest at both G1 and G2/M phases, and the expressions of the WAF1/Cip1 mRNA and protein. We also examined the changes of acetylated isoforms of histone H4. Dose-response and kinetic analysis suggest a close correlation between the level of histone acetylation and the induction of the WAF1/Cip1 expressions. Using several mutant WAF1/Cip1 promoter fragments, we found that the TSA responsive elements are two Sp1 sites at -82 and -69 relative to the transcription start site. These findings indicate that TSA induces the WAF1/Cip1 promoter through the typical Sp1 sites, in a p53-independent fashion. Furthermore, the Sp1-luc plasmid, containing SV40 promoter-derived three consensus Sp1 binding sites, was markedly activated by TSA, compared to the mutant Sp1-luc plasmid. These results demonstrate that transcriptional activation through the Sp1 sites of the WAF1/Cip1 promoter by TSA coincides with induced hyperacetylation of histone H4. PMID- 9405249 TI - A polypeptide derived from mitochondrial dihydrolipoamide succinyltransferase is located on the plasma membrane in skeletal muscle. AB - Dihydrolipoamide succinyltransferase (DLST) is the core-enzyme of 2-oxoglutarate dehydrogenase complex which is located in mitochondria. In this study, several tissues from rat and human were immunostained with an affinity-purified anti-DLST antibody. Of the tissues examined, the plasma membrane of skeletal muscle was immunostained with the antibody besides mitochondria. Furthermore, subcellular fractionation analysis coupled with Western blotting demonstrated that the antigen of the anti-DLST antibody is distributed on the plasma membrane fraction in addition to the mitochondria fraction in skeletal muscle and that it is free from the complex. The molecular weight of the polypeptide bound to the plasma membrane was about 20 kilodaltons (kDa). The polypeptide was purified by immunoprecipitation and its N-terminal amino-acid sequence was determined. The amino-acid sequence exactly corresponded to a part of DLST. Northern blots revealed the presence of mRNA corresponding to the 20 kDa protein. We are the first to report that a mitochondrial protein is also present on the plasma membrane in skeletal muscle as well as in mitochondria. PMID- 9405250 TI - Phosphorylation of A170 stress protein by casein kinase II-like activity in macrophages. AB - A170 is an oxidative stress-inducible protein having a Zinc finger domain, two PEST sequences, and many potential phosphorylation sites for serine/threonine kinases. These structural features suggest that the phosphorylation of A170 affects its function and degradation. We have found that A170 is phosphorylated in cultured murine peritoneal macrophages. In addition, using recombinant A170 proteins, we found two proteins of 40 and 44 kDa with kinase activity in cell extracts using an in-gel kinase assay. We compared the properties of the intrinsic A170 kinases with those of mitogen-activated protein kinase (ERK 2), protein kinase A (PKA), casein kinase II (CK II), and protein kinase C, since their catalytic subunits have molecular masses similar to A170 kinases. ERK 2, CK II, and PKA phosphorylated recombinant A170 as a substrate. The 40 and 44 kDa kinases present in the macrophage extract were similar to alpha and alpha' subunits of CK II in respect to substrate specificity, pharmacological properties, immuno-reactivities, and ubiquitous expression in tissues. PMID- 9405252 TI - Synergism of the ATF/CRE site and GC box in the housekeeping Na,K-ATPase alpha1 subunit gene is essential for constitutive expression. AB - Na,K-ATPase alpha1 subunit gene is constitutively expressed in a wide variety of tissues. Our previous studies revealed that the promoter region between -77 and +17 of the transcription initiation site of the rat Na,K-ATPase alpha1 subunit gene (Atp1a1) is sufficient for the promoter activity. In this region, an ATF/CRE site with an adjacent GC box exists. To elucidate how these sites are involved in the promoter activity, we analyzed effects of point mutations at these sites on transcription by in vitro transcription assays using nuclear extracts prepared from various rat tissues. Mutation at either site resulted in dramatic reduction of the promoter activity in all nuclear extracts, while mutation at both sites did not lead to further reduction. These results indicate that the ATF/CRE site and GC box are both essential for promoter activity and show synergistic activation. Electrophoretic mobility shift assay indicated that Sp1 and/or Sp3 bind to the GC box, and ATF1-CREB heterodimer binds to the ATF/CRE site. Since an element, ATF/CRE site-GC box, is conserved in mammalian Na,K-ATPase alpha1 subunit genes and in other constitutive promoters, we propose that this element is a critical unit for constitutive expression. PMID- 9405251 TI - Bcl-xL overexpression restricts heat-induced apoptosis and influences hsp70, bcl 2, and Bax protein levels in FL5.12 cells. AB - Although several proteins have been identified that can inhibit stress-induced apoptosis, the cytoprotective potential of bcl-xL against heat shock and its ability to alter hsp70 induction is not known. The current study, using control and bcl-xL-overexpressing IL-3-dependent FL5.12 cells, compared the effects of 1 h of acute heat stress (42 degrees C) followed by 1, 4, and 8 h recovery (37 degrees C) on hsp70, bax, bcl-2, and bcl-xL protein levels and apoptosis. Less than 0.5% of untreated cells were apoptotic. There was significantly more apoptosis in control ( approximately 16%) as compared to bcl-xL cells ( approximately 3%) 8 h after heat stress. Immunoblotting revealed a time-dependent increase in hsp70 protein levels following 1 h of heat stress in control, but not bcl-xL-overexpressing cells. bcl-2 protein levels were lower in bcl-xL overexpressing cells than in controls, but decreased in both cell lines after heat stress. bax protein levels in bcl-xL-overexpressing cells were decreased approximately 80% below baseline levels 1 h post heat shock. This decrease was maintained to 8 h. No change in bax protein was observed in control cells up to 8 h post heat shock. These data indicate that bcl-xL overexpression mitigates the effects of acute heat stress so that hsp70 induction is eliminated and apoptosis is prevented. The rapid loss of bax protein following heat stress in bcl-xL overexpressing, but not control, cells may contribute to their resistance to apoptosis. Conversely, the loss of bcl-2 protein following heat stress in control cells may contribute to their susceptibility to apoptosis. PMID- 9405253 TI - Zinc dependent activation of cAMP-specific phosphodiesterase (PDE4A). AB - Cyclic nucleotide phosphodiesterases (PDE), including PDE4A, contain two consensus sequences (HX3HX24-26E) which have been associated with Zn2+ binding and activation with other proteins. This study shows that Zn2+ activates purified recombinant human PDE4A with an EC50 of <1 microM. The EC50 for Mg2+, the generally accepted activating metal ion, is approximately 100 microM. Zn2+ concentrations higher than 5 microM are inhibitory. Mn2+, Co2+ and Ni2+ also activate PDE4A with EC50 values of approximately 2, 3 and 10 microM, respectively. PDE4A binds 65Zn2+ with a Kd of 0.4 microM and approximately 1:1 stoichiometry. Titrations of PDE4A inhibition with Mg2+ and Zn2+ as activating metal ions showed that the competitive inhibitors R-Rolipram, CDP-840, RS-14203 and KF18280 are shifted at least 10-fold to lower potency in the presence of Zn2+. The effect is likely at the site of inhibitor binding as the Km for cAMP in the presence of Mg2+ and Zn2+ is similar (1-3 microM). The Kd of [3H]-R-Rolipram for PDE4A was increased at least 30-fold from 3 nM (with Mg2+) by the presence of Zn2+. The high affinity of Zn2+ for PDE4A indicates that this metal may play a role in the regulation of PDE4A activity. PMID- 9405254 TI - Fluorescence probe study of the lumenal Ca2+ of the sarcoplasmic reticulum vesicles during Ca2+ uptake and Ca2+ release. AB - A limited amount of information is available about the lumenal Ca2+ kinetics of the sarcoplasmic reticulum (SR). Incubation of mag-fura-2AM permitted to incorporate a sufficient amount of the probe into the SR vesicles, as determined by Mn2+ quenching. Rapid changes in the lumenal [Ca2+] ([Ca2+]lum) during Ca2+ uptake and release could be monitored by following the signal derived from the lumenal probe while clamping the extra-vesicular Ca2+ ([Ca2+]ex) at various desired levels with a BAPTA/Ca buffer. Changes in the [Ca2+]lum during uptake and release show the characteristics intrinsic to the SR Ca2+ pump (the [Ca2+]ex dependence of the activation and inhibition by thapsigargin) and the Ca2+ release channel (blocking by ruthenium red), respectively. A new feature revealed by the [Ca2+]lum measurement is that during the uptake reaction the free [Ca2+]lum showed a significant oscillation. Several pieces of evidence suggest that this is due to some interactions between the Ca2+ pump and lumenal proteins. PMID- 9405256 TI - Influence of capping and polyadenylation on mRNA expression and on antisense RNA mediated inhibition of gene expression. AB - In order to investigate the influence of CAP and poly(A)-tail on mRNA expression and on antisense mediated inhibition of gene expression, coinjections of different expression vectors coding for Chloramphenicol-Acetyltransferase (CAT) sense- or antisense RNAs, respectively, were performed. Different in vitro transcribed and modified sense and antisense RNAs were injected into nucleus or cytoplasm of COS7-cells. It can be concluded that the combination of capping and polyadenylation ensures efficient gene expression and is required for transport of mRNA from the nucleus to the cytoplasm. In contrast, antisense experiments suggest that the length of antisense RNAs play an important role for the inhibitory capability of antisense molecules and expression reduction occurs independently of CAP and poly(A) tail. A model for intermolecular hybridization and suppression of gene expression based on the secondary structures of the CAT mRNA and its corresponding antisense RNA is presented. PMID- 9405255 TI - Large unphosphorylated aggregates as the active form of hsp27 which controls intracellular reactive oxygen species and glutathione levels and generates a protection against TNFalpha in NIH-3T3-ras cells. AB - The mammalian small stress protein hsp27 is an oligomeric phosphoprotein which interferes with the cell death induced by several stimuli. In that sense, we and others have recently shown that human hsp27 expression induced cellular protection against tumor necrosis factor (TNFalpha), a protection which depends on the ability of hsp27 to decrease the level of reactive oxygen species and increase that of glutathione. Here, we have analyzed unphosphorylatable mutants of human hsp27 in which serines 15, 78, and 82 were replaced by alanines, glycines, or aspartic acids. Depending on the amino acid which was used to substitute the serine sites, a different pattern of hsp27 structural organization was observed. Alanine substitution generated large hsp27 aggregates while glycine and aspartic acid did the reverse. Hence, these phosphorylatable serine residues can be considered as key elements affecting hsp27 structural organization. Only the large aggregates of hsp27 were able to modulate reactive oxygen species and glutathione and generated cellular protection against TNFalpha. Moreover, using drugs that modulate the intracellular level of glutathione, we show that an increase in glutathione by itself was sufficient to generate large hsp27 structures while the reverse was observed in the case of glutathione deprivation. PMID- 9405257 TI - Genetic analysis of non-insulin-dependent diabetes mellitus in the Otsuka Long Evans Tokushima Fatty rat. AB - The Otsuka Long-Evans Tokushima Fatty (OLETF) rat is an animal model for obese NIDDM. We performed a genome wide scan in F2 progenies obtained by crossing OLETF rats with two control strains, Long-Evans Tokushima Otsuka (LETO) and Fisher 344(F-344) rats. Since diabetes develops only in male progenies, we used only male F2 rats for the linkage studies.Highly significant linkage was observed between the phenotype, postprandial hyperglycemia and P-450ald locus on chromosome 1 and D7Mit 11 locus on chromosome 7. In addition, suggestive linkage was found between fasting glucose level and body weight and these two loci. Four other regions (D1Mit12, D2Mit11, D5Mgh14, and D17Arb1) on chromosome 1, 2, 5, and 17 were detected to influence body weight, fasting glucose level or postprandial hyperglycemia independently. We concluded that non-insulin-dependent diabetes mellitus(NIDDM) in OLETF rats is regulated by multiple genes which affect fasting, postprandial hyperglycemia, and obesity differently. PMID- 9405258 TI - Endothelial derived vasorelaxation is impaired in human APO A-I transgenic rabbits. AB - Endothelium-derived relaxing factor (nitric oxide: NO) may provide an endogenous defence against atherosclerosis which impairs endothelium-dependent vascular relaxation. Atherosclerosis development is inhibited in cholesterol fed human apo A-I transgenic rabbits (Duverger, N., Circulation, 1996, 94, 713-717). We investigated if endothelium-dependent vascular relaxation is modified in human apo A-I transgenic rabbits by testing in vitro endothelium-dependent receptor dependent vascular relaxation to acetylcholine and endothelium-dependent receptor independent vascular relaxation to A23187 of abdominal aorta, precontracted with phenylephrine, in human apo A-I transgenic rabbits (n=4) versus non transgenic littermates (n=4). Endothelium-independent vascular relaxation was investigated with sodium nitroprusside. Vascular precontraction to phenylephrine was significantly increased in human apo A-I transgenic rabbits (p<0.05) while endothelium-independent vascular relaxation to nitroprusside was similar between human apo A-I transgenic rabbits and control rabbits. Endothelium-dependent receptor-dependent and receptor-independent vascular relaxations were reduced in human apo A-I transgenic rabbits (p<0.05). Maximum endothelium-dependent receptor dependent vascular relaxation was negatively correlated with HDL-cholesterol and total apo A-I (rabbit+ human) plasma levels (r=0.87 and 0.86, p=0.01, respectively) but not with atherogenic plasma lipid (VLDL-cholesterol, LDL cholesterol, VLDL+LDL cholesterol, triglycerides, apolipoprotein B) levels. These results suggest that the transgenesis of human apo A-I in rabbits impairs signal transduction of endothelial NO synthesis. PMID- 9405260 TI - Introduction: homeostasis in the immune system AB - No abstractCopyright 1997 Academic Press Limited Copyright 1997Academic Press Limited PMID- 9405259 TI - Upregulation of osteopontin in ischemia-induced renal failure in rats: a role for ET-1? AB - In this study, the involvement of osteopontin in a rat model of ischemia-induced acute renal failure (ARF) was evaluated. In unilaterally nephrectomized Sprague Dawley rats where the left artery was occluded for 30 min., plasma creatinine levels increased significantly within two hours following reperfusion indicating the onset of renal failure. Northern analysis of kidney cortical RNA from these rats showed a time-dependent increase in osteopontin mRNA expression that was significantly higher than sham-operated rats. Since endothelin-1 (ET-1) is implicated as a mediator of acute renal failure, we evaluated its effects on osteopontin expression in a rat mesangial cell-line. Data from in vitro studies indicated that endothelin-1 (ET-1) caused a modest but reproducible increase in osteopontin mRNA in these cells. While the signal for osteopontin upregulation in the rat model is not known, ET-1, which is known to be increased during ischemia, may contribute at least in part to this process. PMID- 9405261 TI - Lymphocyte homeostasis. AB - B- and T-lymphocyte populations have an independent homeostatic regulation of resting (B and T) and activated (B) or memory (T) cell compartments. This organization may provide an efficient mechanism to ensure simultaneously a first natural barrier of protection against common pathogens, the maintenance of immunological T-cell memory and a reservoir of repertoire diversity capable of dealing with new antigenic challenges. PMID- 9405262 TI - Restoration of T-cell homeostasis after T-cell depletion. AB - T-cell homeostasis appears to be maintained throughout much of normal adult life independent of de-novo production from hematopoietic stem cells via thymopoiesis. Instead, peripheral mechanisms are generally sufficient to maintain normal T-cell number, function and adequate TCR repertoire diversity in healthy hosts. Studies of T-cell regeneration in animals, however, have shown that full restoration of T cell homeostasis after profound T-cell depletion is primarily dependent upon thymopoiesis. In this setting, thymic-deficient hosts have prolonged reductions in total T-cell number, restricted TCR repertoire diversity, and limited immunocompetence. In humans, age-related reductions in thymic regenerative capacity as early as young adulthood result in incomplete restoration of T-cell homeostasis after T-cell depletion. PMID- 9405263 TI - The peripheral T-cell pool: regulation by non-antigen induced proliferation? AB - Despite continuous perturbations, the recirculating lymphocyte pool remains relatively constant. Expansion of the pool is compensated for by cell loss. When the T-cell pool is in deficit, either from a congenital defect or an acquired immunodeficiency, T-cell numbers are restored by extrathymic division-a response that occurs without deliberate provocation. We have considered how the recirculating pool may be stably maintained and how a T-cell deficit might be restored to equilibrium. Recent evidence suggests that depleted T-cell compartments are replenished by CD4 T-cell proliferation in the absence of specific antigen, a response that occurs without engaging the T-cell receptor. PMID- 9405264 TI - An immune system switch at birth triggers a change in the lifespan of peripheral T cells. AB - Lymphocyte recirculation is an essential element in the integration of immune responses and is an absolute requirement for the development of systemic memory in postnatal animals. During foetal life a large pool of recirculating T cells develops and migration pathways of naive T cells to skin and peripheral tissues as well as LN are established. At birth a process is triggered whereby naive fetal T cells are rapidly lost from the circulating pool and are replaced by newly arriving T cells which have been formed since birth. At present our data suggest that the thymic export in the fetus creates a pool of long-lived naive T cells of wide diversity. The situation in neonatal lambs is more complex since the thymus is exporting large numbers of short-lived thymic emigrants which enter a peripheral T-cell population where many T cells are dividing. PMID- 9405265 TI - Consequences of viral infections for lymphocyte compartmentalization and homeostasis. AB - The immune system has evolved to deal with pathogens. Analysing what happens during the course of infectious processes provides insights into the limits of lymphocyte homeostasis. Virus infections greatly alter normal T- and B-cell prevalence and localization patterns. Any mechanism that 'counts' T cells and B cells seems to be disrupted, at least while antigen persists. There is no simple 'dumping' process that controls numbers in the blood. Though the cell-surface 'language' that determines lymphocyte trafficking patterns must be central to modulating the consequences of infectious diseases, it is far from clear how such interactions maintain the system in reasonable balance. PMID- 9405266 TI - Homeostatic mechanisms in the control of autoimmunity. AB - The data in this article challenge the current paradigm that self tolerance among T cells can be fully accounted for in terms of intrathymic clonal deletion, anergy and T-cell 'indifference'. We show that autoreactive T cells are under the control of a dominant T-cell-mediated control mechanism and that the thymus is a particularly potent source of these protective cells. We further show that 'peripheral' self antigens implicated in autoimmune diseases are expressed intrathmically and we consider possible consequences of such expression. Finally we show that autoantigen-specific peripheral T cells can be programmed to prevent autoimmunity. PMID- 9405267 TI - T-cell homeostasis in HIV-1 infection. AB - Failure of T-cell homeostasis is an important feature of HIV-1 infection. Substantial evidence indicates that T-cell homeostasis is independent of CD4+ and CD8+ subsets, and this may contribute to the decline of CD4+ T cells to low levels in this disease. Moreover, failure of T-cell homeostasis appears to precede the development of clinically-defined AIDS by approximately 1.5 to 2 years and is thus an important milestone in HIV-1 disease progression. We argue that T-cell turnover and depletion of memory cells in HIV-1 infection can be viewed as the reverse of the process by which immune reconstitution occurs after stem cell transplantation, and that changes in the functional level of T-cell memory may be critical to both processes. An understanding of the relationship between T-cell memory and regeneration of lost T cells may help preserve and/or reconstitute immune system homeostasis in HIV-1-infected individuals. PMID- 9405268 TI - Dynamics of fine T-cell subsets during HIV disease and after thymic ablation by mediastinal irradiation. AB - The T-cell compartment is considerably more complex than just CD4 and CD8 T cells. Indeed, we can identify dozens of functionally and phenotypically distinct subsets within the peripheral blood of humans. These subsets are differentially affected in diseases which may underly some of the functional defects attributable to the disease. In HIV disease, all thymic-derived T-cell populations are gradually lost at identical rates during late-stage disease progression, while unusual, perhaps extrathymically-derived T cells expand. This expansion may reflect an attempt on the part of the immune system to compensate for the significant insult of HIV infection to the host: the abrogation of normal thymopoiesis and T-cell homeostasis. PMID- 9405269 TI - Thymic function in HIV-1 disease. AB - The thymus involutes over time and is generally considered to be non-functional in adults. If this is the case, T-cell depletion in adult HIV-1 disease would in part be due to insufficient intrathymic production. This review summarizes information relevant to the structure and function of the thymus under normal conditions and in the setting of infection with HIV-1. The speculation is raised: could residual thymic reserve be present in some adults and be inducible during pathophysiological states of T-cell depletion? If so, the contribution of thymopoiesis during adult HIV-1 disease may be more significant than otherwise assumed. PMID- 9405272 TI - Cytosolic aspartate aminotransferase gene is a member of the glucose-regulated protein gene family in adipocytes. AB - Stress controls the expression of a cohort of genes. Among these, the glucose regulated protein (GRP) genes are specifically activated by glucose deprivation, reducing agents, glycosylation block, intracellular calcium or ex vivo incubations of tissues or cells. We demonstrate that these stimuli induce the expression of the cytosolic aspartate aminotransferase gene in adipocytes by a process involving the region of the promoter between -2405 and -26 bp. Therefore this transaminase is a new member of the GRP family. PMID- 9405273 TI - Phospholipase A2 from bovine seminal plasma is a platelet-activating factor acetylhydrolase. AB - The major phospholipase A2 activity from bovine seminal plasma was recently purified [Soubeyrand, Khadir, Brindle and Manjunath (1997) J. Biol. Chem. 272, 222-227]. We here show that the 60 kDa enzyme is in fact a platelet-activating factor acetylhydrolase (PAF-AH). Sequences of the N-terminus as well as of internal fragments showed 100% identity with the cDNA-deduced sequences of bovine plasma PAF-AH. The enzyme has kinetic properties similar to those of the human serum PAF-AH. Although capable of hydrolysing long-chained phosphatidylcholine, it displayed a highly preferential activity towards PAF. The enzyme activity towards phosphatidylcholine, but not PAF, was Ca2+-dependent. Biochemical characterization revealed that the enzyme is extensively N-glycosylated and that it exists predominantly as a dimer in solution. Western blot analysis revealed that the enzyme is highly heterogeneous in charge, with a maximal distribution at an isoelectric point of approx. 5.7. The enzyme was expressed exclusively in the seminal vesicles and the ampulla. No association of the enzyme with either epididymal or ejaculated spermatozoa could be detected. PMID- 9405271 TI - Zonation of hepatic cytochrome P-450 expression and regulation. AB - The CYP genes encode enzymes of the cytochrome P-450 superfamily. Cytochrome P 450 (CYP) enzymes are expressed mainly in the liver and are active in mono oxygenation and hydroxylation of various xenobiotics, including drugs and alcohols, as well as that of endogenous compounds such as steroids, bile acids, prostaglandins, leukotrienes and biogenic amines. In the liver the CYP enzymes are constitutively expressed and commonly also induced by chemicals in a characteristic zonated pattern with high expression prevailing in the downstream perivenous region. In the present review we summarize recent studies, mainly based on rat liver, on the factors regulating this position-dependent expression and induction. Pituitary-dependent signals mediated by growth hormone and thyroid hormone seem to selectively down-regulate the upstream periportal expression of certain CYP forms. It is at present unknown to what extent other hormones that also affect total hepatic CYP activities, i.e. insulin, glucagon, glucocorticoids and gonadal hormones, act zone-specifically. The expression and induction of CYP enzymes in the perivenous region probably have important toxicological implications, since many CYP-activated chemicals cause cell injury primarily in this region of the liver. PMID- 9405274 TI - Regulation of the serine-base exchange enzyme system by CD4: effects of monoclonal antibodies, jacalin, interleukin 16 and the HIV membrane protein gp120. AB - Phosphatidylserine (PtdSer) is synthesized by an exchange of the polar head group of phospholipids for a serine residue. The enzyme responsible for this reaction, the serine-base exchange enzyme system (serine-BEES) is inhibited during lymphocyte activation. We show here that triggering the CD4 cell surface molecule in several CD4+ T-cell lines regulates the serine-BEES activity, thus resulting in marked changes in PtdSer synthesis. CD4 ligands able to generate an activating signal in T-cells such as the lectin jacalin, down-regulate the synthesis of PtdSer. In contrast, monoclonal antibodies (mAbs) directed against the CD4 molecule, such as IOT4 and IOT4a, which have previously been described as generating an inhibitory signal to T-cells, induced an up-regulation of the serine-BEES and impaired CD3-induced inhibition of PtdSer synthesis. Similarly, the HIV-gp120 envelope glycoprotein, in both soluble and cross-linked forms, induces an increase in PtdSer synthesis. The protein tyrosine kinase p56lck participates in the regulation of serine-BEES activity because the effect of CD4 mAbs was additive to that of amino-hydroxyflavone, an inhibitor of p56lck. Also, CD4 mAbs were inactive in J Cam 1.6 cells or when the CD3 signals were bypassed by using thapsigargin. These results demonstrate that the CD4 surface molecule can transmit both activating and inhibiting intracellular signals depending on the CD4 ligand used. We suggest that PtdSer synthesis would be one of the intracellular signals that could explain the opposite effects of different CD4 ligands on T-cells. PMID- 9405275 TI - Binding of urokinase-type plasminogen activator-plasminogen activator inhibitor-1 complex to the endocytosis receptors alpha2-macroglobulin receptor/low-density lipoprotein receptor-related protein and very-low-density lipoprotein receptor involves basic residues in the inhibitor. AB - The complex of the type-1 plasminogen activator inhibitor (PAI-1) and its target proteinases, the urokinase and tissue-type plasminogen activators (uPA and tPA), but not the free components, bind with high affinity to the endocytosis receptors alpha2-macroglobulin receptor/low-density lipoprotein receptor-related protein (alpha2MR/LRP) and very-low-density lipoprotein receptor (VLDLR). To characterize the molecular interaction between the complexes and the receptors, alanine codons were introduced into the human PAI-1 cDNA to replace the four basic residues, Arg 78, Lys-82, Arg-120 and Lys-124, as double mutations. The purified recombinant mutant proteins, rPAI-1/R78A-K124A and rPAI-1/K82A-R120A, produced by the yeast Pichia pastoris, were indistinghuisable from wild-type recombinant and natural human PAI-1 with respect to inhibitory activity against uPA, stability of SDS resistant complexes with uPA, and vitronectin binding. Radiolabelled mutant uPA.PAI-1 complexes bound with a 10- to 20-fold, and 3- to 7-fold reduced affinity to purified alpha2MR/LRP and VLDLR respectively. alpha2MR/LRP-mediated endocytosis of the mutant complexes by COS-1 cells was reduced to 48 and 38% of the level of endocytosis of wild-type PAI-1. Binding of the mutant complexes to the uPA receptor was not affected. These findings suggest that the binding mode of the uPA.PAI-1 complex to both alpha2MR/LRP and VLDLR is similar. The four residues are surface exposed in the region defined by alpha-helix D and beta strand 1A in the serine protease inhibitor (serpin) structure. Our study represents the first identification of residues in a surface region implicated in molecular recognition of protease.serpin complexes by endocytosis receptors of the low-density lipoprotein receptor family. PMID- 9405276 TI - Expression of the Schwanniomyces occidentalis SWA2 amylase in Saccharomyces cerevisiae: role of N-glycosylation on activity, stability and secretion. AB - The role of N-linked glycosylation on the biological activity of Schwanniomyces occidentalis SWA2 alpha-amylase, as expressed in Saccharomyces cerevisiae, was analysed by site-directed mutagenesis of the two potential N-glycosylation sites, Asn-134 and Asn-229. These residues were replaced by Ala or Gly individually or in various combinations and the effects on the activity, secretion and thermal stability of the enzyme were studied. Any Asn-229 substitution caused a drastic decrease in activity levels of the extracellular enzyme. In contrast, substitutions of Asn-134 had little or no effect. The use of antibodies showed that alpha-amylase was secreted in all the mutants tested, although those containing substitutions at Asn-229 seemed to have a lower rate of synthesis and/or higher degradation than the wild-type strain. alpha-Amylases with substitution at Asn-229 had a 2 kDa lower molecular mass than the wild-type protein, as did the wild-type protein itself after treatment with endoglycosidase F. These findings indicate that Asn-229 is the single glycosylated residue in SWA2. Thermostability analysis of both purified wild-type (T50=50 degrees C, where T50 is the temperature resulting in 50% loss of activity) and mutant enzymes indicated that removal of carbohydrate from the 229 position results in a decrease of approx. 3 degrees C in the T50 of the enzyme. The Gly-229 mutation does not change the apparent affinity of the enzyme for starch (Km) but decreases to 1/22 its apparent catalytic efficiency (kcat/Km). These results therefore indicate that glycosylation at the 229 position has an important role in the extracellular activity levels, kinetics and stability of the Sw. occidentalis SWA2 alpha-amylase in both its wild-type and mutant forms. PMID- 9405277 TI - Evidence for altered sensitivity of the nitric oxide/cGMP signalling cascade in insulin-resistant skeletal muscle. AB - Nitric oxide activates guanylate cyclase to form cGMP, comprising a signalling system that is believed to be a distinct mechanism for increasing glucose transport and metabolism in skeletal muscle. The effects of a selective cGMP phosphodiesterase inhibitor, zaprinast, on basal glucose utilization was investigated in incubated rat soleus muscle preparations isolated from both insulin-sensitive (lean Zucker; Fa/?) and insulin-resistant (obese Zucker; fa/fa) rats. Zaprinast at 27 microM significantly increased cGMP levels in incubated soleus muscle isolated from lean, but not obese, Zucker rats. Muscles were incubated with 14C-labelled glucose and various concentrations of zaprinast (3, 27 and 243 microM). Zaprinast (at 27 and 243 microM) significantly increased rates of net and 14C-labelled lactate release and of glycogen synthesis in lean Zucker rat soleus muscle; glucose oxidation was also increased by 27 microM zaprinast. In addition, regardless of concentration, the phosphodiesterase inhibitor failed to increase any aspect of 14C-labelled glucose utilization in soleus muscles isolated from obese Zucker rats. The maximal activity of nitric oxide synthase (NOS) was significantly decreased in insulin-resistant obese Zucker muscles. Thus the lack of effect of zaprinast in insulin-resistant skeletal muscle is consistent with decreased NOS activity. To test whether there is a defect in insulin-resistant skeletal muscle for endogenous activation of guanylate cyclase, soleus muscles were isolated from both insulin-sensitive and insulin-resistant Zucker rats and incubated with various concentrations of the NO donor sodium nitroprusside (SNP; 0.1, 1, 5 and 15 mM). SNP significantly increased rates of net and 14C-labelled lactate release, as well as glucose oxidation in muscles isolated from both insulin-sensitive and insulin-resistant rats. A decreased response to SNP was observed in the dose-dependent generation of cGMP within isolated soleus muscles from insulin-resistant rats. A possible link between impaired NO/cGMP signalling and abnormal glucose utilization by skeletal muscle is discussed. PMID- 9405279 TI - Studies of the long-term regulation of hepatic pyruvate dehydrogenase kinase. AB - The administration of a low-carbohydrate/high-saturated-fat (LC/HF) diet for 28 days or starvation for 48 h both increased pyruvate dehydrogenase kinase (PDHK) activity in extracts of rat hepatic mitochondria, by approx. 2.1-fold and 3.5 fold respectively. ELISAs of extracts of hepatic mitochondria, conducted over a range of pyruvate dehydrogenase (PDH) activities, revealed that mitochondrial immunoreactive PDHKII (the major PDHK isoform in rat liver) was significantly increased by approx. 1.4-fold after 28 days of LC/HF feeding and by approx. 2 fold after 48 h of starvation. The effect of LC/HF feeding to increase hepatic PDHK activity was retained through hepatocyte preparation, but was decreased on 21 h culture with insulin (100 micro-i.u./ml). A sustained (24 h) 2-4-fold elevation in plasma insulin concentration in vivo (achieved by insulin infusion via an osmotic pump) suppressed the effect of LC/HF feeding so that hepatic PDHK activities did not differ significantly from those of (insulin-infused) control rats. The increase in hepatic PDHK activity evoked by 28 days of LC/HF feeding was prevented and reversed (within 24 h) by the replacement of 7% of the dietary lipid with long-chain omega-3 fatty acids. Analysis of hepatic membrane lipid revealed a 1.9-fold increase in the ratio of total polyunsaturated omega-3 fatty acids to total mono-unsaturated fatty acids. The results indicate that the increased hepatic PDHK activities observed in livers of LC/HF-fed or 48 h-starved rats are associated with long-term actions to increase hepatic PDHKII concentrations. The long-term regulation of hepatic PDHK by LC/HF feeding might be achieved through an impaired action of insulin to suppress PDHK activity. In addition, the fatty acid composition of the diet, rather than the fat content, is a key influence. PMID- 9405278 TI - Effect of reduced low-density lipoprotein receptor level on HepG2 cell cholesterol metabolism. AB - Low-density lipoproteins (LDL) are taken up by both LDL receptor (LDLr)-dependent and -independent pathways. In order to determine the importance of these pathways in the activity of the various enzymes that are important in maintaining the cellular cholesterol level in hepatic cells, we created HepG2 cells expressing lower levels of LDLr. Thus HepG2 cells were transfected with a constitutive expression vector (pRc/CMV) containing a fragment of LDLr cDNA inserted in an antisense manner. Stable transformants were obtained that showed significant reductions of 42, 72 and 85% of LDLr protein levels compared with the control, as demonstrated by immunoblotting and confirmed by the LDL binding assay. The best inactivation was achieved with the construct containing the first 0.7 kb of LDLr cDNA. Incubating the different HepG2 cell subtypes with LDL showed similar association of apolipoprotein B (apo B) or cholesteryl esters from LDL with the cells, indicating that the LDLr deficiency did not significantly affect LDL uptake by the cell. However, apoB degradation was reduced significantly by 71-82% in the most LDLr-deficient HepG2 cells. We also found that 3-hydroxy-3 methylglutaryl-CoA reductase (HMGCoA red) activity is significantly increased by 32-35% in HepG2 cells expressing very low levels of LDLr that also demonstrate a significant decrease of 20% in acyl-CoA:cholesterol acyltransferase (ACAT) activity. However, these effects are moderate compared with those observed when cells were incubated in lipoprotein-depleted medium, where a >900% increase in HMGCoA red activity and a loss of 60% of ACAT activity was observed. Thus, in HepG2 cells, different levels of LDLr affect LDL-apoB degradation, but have very little effect on LDL association, HMGCoA red and ACAT activities, revealing that LDLr is more important in the clearance of LDL-apoB than in HepG2 cell cholesterol homoeostasis, a role that should be attributable to both LDLr dependent and -independent pathways. PMID- 9405280 TI - Importance of the redox state of cytochrome c during caspase activation in cytosolic extracts. AB - The export of cytochrome c from mitochondria to the cytoplasm has been detected during apoptosis. Addition of cytochrome c to cytosolic extracts can activate the caspases, suggesting that this export could be an important intracellular signal for initiating the apoptotic programme. We have investigated the mechanism of caspase activation by cytochrome c. Mitochondrial cytochrome c normally shuttles electrons between complexes III and IV of the electron transport chain. Interaction with these complexes is dependent on electrostatic interactions via a polylysine binding pocket. Cytosolic caspase activation was only observed with intact holocytochrome c, and increasing the ionic composition of the extracts prevented activation, suggesting that stringent allosteric interactions between cytochrome c and other cytoplasmic factors are necessary. Cytochrome c was fully reduced within 5 min of addition to the cytosolic extracts. Potassium ferricyanide could maintain cytochrome c in an oxidized state, but care was taken to use ferricyanide at concentrations where its polyanion effect did not cause interference. The oxidized form of cytochrome c was able to activate the caspases. We conclude that reduced cytochrome c will function in the cytoplasm; however, its reduction is not a critical step, and electron transfer from cytochrome c to its cytoplasmic-binding partner(s) is not necessary in the pathway leading to apoptosis. PMID- 9405281 TI - Structure of the trigonal crystal form of bovine annexin IV. AB - The structure of a trigonal crystal form of N-terminally truncated [des-(1-9)] bovine annexin IV, an annexin variant that exhibits the distinctive property of binding both phospholipids and carbohydrates in a Ca2+-dependent manner, has been determined at 3 A (0.3 nm) resolution -space group: R3; cell parameters: a=b=118.560 (8) A and c=82.233 (6) A-. The overall structure of annexin IV, crystallized in the absence of Ca2+ ions, is highly homologous to that of the other known members of the annexin family. The trimeric assembly in the trigonal crystals of annexin IV is quite similar to that found previously in non isomorphous crystals of human, chicken and rat annexin V and to the subunit arrangement in half of the hexamer of hydra annexin XII. Moreover, it resembles that found in two-dimensional crystals of human annexin V bound to phospholipid monolayers. The propensity of several annexins to generate similar trimeric arrays supports the hypothesis that trimeric complexes of such annexins, including annexin IV, may represent the functional units that interact with membranes. PMID- 9405282 TI - Differential routes of Ca2+ influx in Swiss 3T3 fibroblasts in response to receptor stimulation. AB - Ca2+ influx into cells in response to stimulation of various receptors was studied with Swiss 3T3 fibroblasts. The mechanisms involved were found to be so diverse that they were classified into four groups, Type I to IV. Type-I influx occurred, via pertussis toxin-susceptible G-proteins, immediately after internal Ca2+ mobilization by bradykinin, thrombin, endothelin, vasopressin or angiotensin II. Type-II influx induced by bombesin differed from Type I in its insusceptibility to pertussis toxin treatment. Ca2+ influx induced by prostaglandin E1, referred to as Type-III influx, was unique in that phospholipase C was apparently not activated without extracellular Ca2+, strongly suggesting that the Ca2+ influx preceded and was responsible for InsP3 generation and internal Ca2+ mobilization. More Ca2+ entered the cells more slowly via the Type-IV route opened by platelet-derived and other growth factors. These types of Ca2+ influx could be differentiated by their different susceptibilities to protein kinase C maximally activated by 1 h of exposure of cells to PMA, which inhibited phospholipase Cbeta coupled to receptors involved in Type-I and -II influx but did not inhibit growth-factor-receptor-coupled phospholipase Cgamma. Type-I and -II Ca2+ influxes, together with store-operated influx induced by thapsigargin, were not directly inhibited by exposure of cells to PMA, but Type III and -IV influxes were completely inhibited. In addition, stimulation of receptors involved in Type-I and -IV Ca2+ influx, but not Type-II and -III influx, led to phospholipase A2 activation in the presence of extracellular Ca2+. Inhibition of Type-I and -IV Ca2+ influxes by their respective inhibitors, diltiazem and nifedipine, resulted in abolition of phospholipase A2 activation induced by the respective receptor agonists, in agreement with the notion that Ca2+ influx via these routes is responsible for receptor-mediated phospholipase A2 activation. PMID- 9405283 TI - Regulation of PtdIns4P 5-kinase C by thrombin-stimulated changes in its phosphorylation state in human platelets. AB - PtdIns(4,5)P2 production by the enzyme PtdIns4P 5-kinase C (PIPkin C) was examined in thrombin-stimulated human platelets. Thrombin caused a rapid, transient 2-3-fold increase in PIPkin activity and a transient net dephosphorylation of the enzyme. PIPkin C was phosphorylated on serine and threonine residues in unstimulated platelets; no evidence for tyrosine phosphorylation was found. The phosphatase inhibitor okadaic acid promoted PIPkin C hyperphosphorylation and a concomitant marked inhibition of its activity in immunoprecipitates. Activity was restored by treatment with alkaline phosphatase, suggesting the existence of an inhibitory phosphorylation site. In support of this idea, alkaline phosphatase treatment of PIPkin C immunoprecipitated from unstimulated platelets caused a modest (1.6-fold) but significant activation of the enzyme. However, alkaline phosphatase treatment of PIPkin C immunoprecipitated from thrombin-stimulated platelets caused a decrease in activity to approximately the same levels, suggesting that the phosphorylation of PIPkin C also contributes to the observed stimulation. Two-dimensional phosphopeptide mapping of immunoprecipitated PIPkin C revealed that the enzyme is multiply phosphorylated and that, whereas some phosphopeptides are indeed lost on stimulation, consistent with the net dephosphorylation of the enzyme, at least two novel sites become phosphorylated. This suggests that thrombin causes complex changes in the phosphorylation state of PIPkin C, one consequence of which is its activation. PMID- 9405284 TI - Comparison of the roles of mitogen-activated protein kinase kinase and phosphatidylinositol 3-kinase signal transduction in neutrophil effector function. AB - Although it is known that many stimuli can activate mitogen-activated protein kinases (MAPKs) and phosphatidylinositol 3-kinases (PI3K) in human neutrophils, little is known concerning either the mechanisms or function of this activation. We have utilized a selective inhibitor of MAPK kinase (MEK), PD098059, and two inhibitors of PI3K, wortmannin and LY294002, to investigate the roles of these kinases in the regulation of neutrophil effector functions. Granulocyte/macrophage colony-stimulating factor, platelet-activating factor (PAF) and N-formylmethionyl-leucyl-phenylalanine are capable of activating both p44ERK1 and p42ERK2 MAPKs and phosphotyrosine-associated PI3K in human neutrophils. The activation of extracellular signal-related protein kinases (ERKs) is correlated with the activation of p21ras by both tyrosine kinase and G protein-coupled receptors as measured by a novel assay for GTP loading. Wortmannin and LY294002 inhibit, to various degrees, superoxide generation, neutrophil migration and PAF release. Incubation with PD098059, however, inhibits only the PAF release stimulated by serum-treated zymosan. This demonstrates that, while neither MEK nor ERK kinases are involved in the activation of respiratory burst or neutrophil migration, inhibition of PAF release suggests a potential role in the activation of cytosolic phospholipase A2. PI3K isoforms, however, seem to have a much wider role in regulating neutrophil functioning. PMID- 9405285 TI - Cross-talk between transcriptional regulation by thyroid hormone and myogenin: new aspects of the Ca2+-dependent expression of the fast-type sarcoplasmic reticulum Ca2+-ATPase. AB - We have previously demonstrated an interaction between the major determinants of skeletal muscle phenotype by showing that continuous contractile activity represses the thyroid hormone (3,3', 5-tri-iodothyronine; T3)-dependent transcriptional activity of fast-type sarcoplasmic/endoplasmic-reticulum Ca2+ ATPase (SERCA1), a characteristic of the fast phenotype. Both the free cytosolic Ca2+ concentration ([Ca2+]i) and the myogenic determination factors MyoD and myogenin have been implicated as mediators of the effect of contractile activity on skeletal muscle phenotype. Using L6 cells we have shown that an increase in the steady-state [Ca2+]i above the resting level of 120 nM indeed can mimic the effect of contractile activity on T3-dependent SERCA1 expression. We now show that the repressing effect of increased [Ca2+]i on T3-dependent SERCA1 expression in L6 cells is exerted at a pre-translational level and is accompanied by increased myogenin mRNA expression. Myogenin overexpression in these cells revealed that increased expression of myogenin alone strongly decreases the T3 dependent stimulation of SERCA1 promoter activity. These results suggest a pathway for the regulation of skeletal muscle phenotype in which [Ca2+]i mediates the effect of contractile activity by regulating the expression of myogenin, which in turn interferes with transcriptional regulation by T3. PMID- 9405286 TI - Role of non-compatible osmolytes in the stabilization of proteins during heat stress. AB - This study is a systematic attempt to understand the roles of non-compatible osmolytes, i.e. solutes that have inhibitory effects on enzymes, in the stabilization of proteins against denaturing stress. Thermal denaturation of RNase A, holo-alpha-lactalbumin, apo-alpha-lactalbumin, lysozyme and metmyoglobin in the absence and presence of various concentrations of free basic amino acids was studied by observing changes in the absorption coefficients of these proteins. It has been observed that arginine and histidine destabilize all proteins in terms of the midpoint of the transition curve and Gibbs energy change on denaturation. Study of the heat-induced denaturation of the proteins in the presence of various concentrations of arginine at different pH values demonstrated that arginine binds to the denatured molecules. In contrast with the effect of arginine and histidine on protein stability, it was observed that the effect of lysine on proteins stability is unpredictable, i.e. it may have a stabilizing effect, no effect or a destabilizing effect on proteins during denaturing stress. The results of this study are considered from an evolutionary perspective. PMID- 9405287 TI - Recombinant human sulphamidase: expression, amplification, purification and characterization. AB - Mucopolysaccharidosis type IIIA (MPS IIIA, Sanfilippo A syndrome) is a lysosomal storage disease that causes a profound neurological deterioration. The disorder is caused by a deficiency of the lysosomal enzyme sulphamidase which is a requisite for the degradation of heparan sulphate. To facilitate the development of enzyme-replacement strategies for MPS IIIA patients, we have constructed a high-level expression system for recombinant human sulphamidase in Chinese hamster ovary (CHO) cells. An expression construct containing a methotrexate resistant dihydrofolate reductase (DHFR) gene allowed amplification of expression levels from less than 1 mg of sulphamidase per litre of culture medium to approx. 15 mg/l. Unlike many cell lines made by gene amplification in DHFR-deficient CHO cells, and utilizing the normal DHFR gene, these cell lines appeared to be stable in the absence of selective pressure. Recombinant human sulphamidase was purified from unamplified and amplified cell lines. The native enzyme was found to be a dimer of 115 kDa. Denaturing and reducing SDS/PAGE revealed a subunit size of 62 kDa. Kinetic analysis demonstrated that the recombinant enzyme had broadly similar kinetic characteristics to sulphamidase purified from liver. Recombinant human sulphamidase was able to correct the storage phenotype of MPS IIIA fibroblasts after endocytosis via the mannose-6-phosphate receptor. PMID- 9405288 TI - Enzymic aromatization of 6-alkyl-substituted androgens, potent competitive and mechanism-based inhibitors of aromatase. AB - To gain insight into the relationships between the aromatase inhibitory activity of 6-alkyl-substituted androgens, potent competitive inhibitors, and their ability to serve as a substrate of aromatase, we studied the aromatization of a series of 6alpha- and 6beta-alkyl (methyl, ethyl, n-propyl, n-pentyl and n heptyl)-substituted androst-4-ene-3,17-diones (ADs) and their androsta-1,4-diene 3,17-dione (ADD) derivatives with human placental aromatase, by gas chromatography-mass spectrometry. Among the inhibitors examined, ADD and its 6alpha-alkyl derivatives with alkyl functions less than three carbons long, together with 6beta-methyl ADD, are suicide substrates of aromatase. All of the steroids, except for 6beta-n-pentyl ADD and its n-heptyl analogue as well as 6beta-n-heptyl AD, were found to be converted into the corresponding 6-alkyl oestrogens. The 6-methyl steroids were aromatized most efficiently in each series, and the aromatization rate essentially decreased in proportion to the length of the 6-alkyl chains in each series, where the 6alpha-alkyl androgens were more efficient substrates than the corresponding 6beta isomers. The Vmax of 6alpha-methyl ADD was approx. 2.5-fold that of the natural substrate AD and approx. 3-fold that of the parent ADD. On the basis of this, along with the facts that the rates of a mechanism-based inactivation of aromatase by ADD and its 6alpha-methyl derivative are similar, it is implied that alignment of 6alpha methyl ADD in the active site could favour the pathway leading to oestrogen over the inactivation pathway, compared with that of ADD. The relative apparent Km values for the androgens obtained in this study are different from the relative Ki values obtained previously, indicating that there is a difference between the ability to serve as an inhibitor and the ability to serve as a substrate in the 6 alkyl androgen series. PMID- 9405289 TI - Ca2+ administration stimulates the binding of AP-1 factor to the 5'-flanking region of the rat gene for the Ca2+-binding protein regucalcin. AB - mRNA of the Ca2+-binding protein, regucalcin, is mainly expressed in the liver and only to a small extent in the kidney, and the expression of hepatic regucalcin mRNA is markedly stimulated by Ca2+ administration [Shimokawa and Yamaguchi (1992) FEBS Lett. 305, 151-154]. The existence of nuclear factors that bind to the 5'-flanking region of the rat regucalcin gene was investigated. When nuclear proteins obtained from various rat tissues were used in gel mobility shift assays, tissue-specific formation of a protein-DNA complex was found in the liver and kidney. An additional novel protein-DNA complex was formed when liver nuclear extracts obtained from Ca2+-administered rats (10mg of Ca2+/100g body weight) were used. Competition gel mobility-shift experiments using consensus and mutant oligonucleotides for AP-1 factor showed that the additional novel complex was formed from binding of the AP-1 factor to the regucalcin gene. Ca2+-induced binding of the AP-1 factor to the regucalcin gene was completely inhibited by simultaneous administration of trifluoperazine, an antagonist of calmodulin, suggesting that the activation of nuclear AP-1 protein is partly mediated through a Ca2+/calmodulin-dependent pathway. Moreover, the 5'-flanking region of the rat regucalcin gene ligated to a luciferase reporter gene possessed the promoter activity in H4-II-E hepatoma cells. This promoter activity was enhanced by treatment with Bay K 8644, a Ca2+-channel agonist. The present study demonstrates that the Ca2+-response sequences are located within the 5'-flanking region of the rat regucalcin gene. PMID- 9405291 TI - Kinetic and chemical mechanisms for the effects of univalent cations on the spectral properties of aromatic amine dehydrogenase. AB - Univalent cations and pH influence the UV-visible absorption spectrum of the tryptophan tryptophylquinone (TTQ) enzyme, aromatic amine dehydrogenase (AADH). Little spectral perturbation was observed when pH was varied in the absence of univalent cations. The addition of alkali metal univalent cations (K+, Na+, Li+, Rb+ or Cs+) to oxidized AADH caused significant changes in its absorption spectrum. The apparent Kd for each cation, determined from titrations of the spectral perturbation, decreased with increasing pH. Transient kinetic studies involving rapid mixing of AADH with cations and pH jump revealed that the rate of the cation-induced spectral changes initially decreased with increasing cation concentration to a minimum value, then increased with increasing cation concentration. A kinetic model was developed to fit these data, determine the true pH-independent Kd values for K+ and Na+, and explain the pH-dependence of the apparent Kd. A chemical reaction mechanism, based on the kinetic data, is presented in which the metallic univalent cation facilitates the chemical modification of the TTQ prosthetic group to form an hydroxide adduct which gives rise to the spectral change. Addition of NH4(+)/NH3 to AADH caused changes in the absorption spectrum which were very different form those caused by addition of the metallic univalent cations. The kinetics of the reaction induced by addition of NH4+/NH3 were also different, being simple saturation kinetics. Another reaction mechanism is proposed for the NH4+/NH3-induced spectral change that involves nucleophilic addition of the unprotonated NH3 to TTQ. The general relevance of these data and models to the physiological reactions of TTQ dependent enzymes and to the roles of univalent cations in modulating enzyme activity are discussed. PMID- 9405290 TI - Characterization of the regulatory regions in the human desmoglein genes encoding the pemphigus foliaceous and pemphigus vulgaris antigens. AB - The adhesive proteins in the desmosome type of cell junction consist of two members of the cadherin superfamily, the desmogleins and desmocollins. Both desmogleins and desmocollins occur as at least three different isoforms with various patterns of expression. The molecular mechanisms controlling the differential expression of the desmosomal cadherin isoforms are not yet known. We have begun an investigation of desmoglein gene expression by cloning and analysing the promoters of the human genes coding for the type 1 and type 3 desmogleins (DSG1 and DSG3). The type 1 isoform is restricted to the suprabasal layers of the epidermis and is the autoantigen in the autoimmune blistering skin disease pemphigus foliaceous. The type 3 desmoglein isoform is also expressed in the epidermis, but in lower layers than the type 1 isoform, and is the autoantigen in pemphigus vulgaris. Phage lambda genomic clones were obtained containing 4.2 kb upstream of the translation start site of DSG1 and 517 bp upstream of the DSG3 start site. Sequencing of 660 bp upstream of DSG1 and 517 bp upstream of DSG3 revealed that there was no obvious TATA box, but a possible CAAT box was present at -238 in DSG1 and at -193 in DSG3 relative to the translation start site. Primer extension analysis and RNase protection experiments revealed four putative transcription initiation sites for DSG1 at positions -163, -151, 148 and -141, and seven closely linked sites for DSG3, the longest being at -140 relative to the translation start site. The sequences at these possible sites at 166 to -159 in DSG1 (TTCAGTCC) and at -124 to -117 in DSG3 (CTTAGACT) have some similarity to the initiator sequence (CTCANTCT) described for a TATA-less promoter often from -3 to +5, and the true transcription initiator site might therefore be the A residue in these sequences. There were two regions of similarity between the DSG1 and DSG3 promoters just upstream of the transcription initiation sites, of 20 and 13 bp, separated by 41 bp in DSG1 and 36 bp in DSG3. The significance of these regions of similarity remains to be elucidated, but the results suggest that they represent a point at which these two desmoglein genes are co-ordinately regulated. Analysis of the upstream sequences revealed GC-rich regions and consensus binding sites for transcription factors including AP-1 and AP-2. Exon boundaries were conserved compared with the classical cadherin E cadherin, but the equivalent of the second cadherin intron was lacking. A 4.2 kb region of the human DSG1 promoter sequence was linked to the lacZ gene reporter gene in such a way that there was only one translation start site, and this construct was used to generate transgenic mice. We present the first transgenic analysis of a promoter region taken from a desmosomal cadherin gene. Our results suggest that the 4.2 kb upstream region of DSG1 does not contain all the regulatory elements necessary for correct expression of this gene but might have elements that regulate activity during hair growth. PMID- 9405292 TI - Soluble form of complement C3b/C4b receptor (CR1) results from a proteolytic cleavage in the C-terminal region of CR1 transmembrane domain. AB - The complement C3b/C4b receptor (CR1) is an integral protein, anchored in the plasma membrane through a hydrophobic domain of 25 amino acids, but is also found in the plasma in soluble form (sCR1). A recombinant, soluble form of CR1 has been demonstrated to reduce complement-dependent tissue injury in animal models of ischaemia/reperfusion. In view of the important pathophysiological relevance of sCR1, we have investigated the mechanisms governing CR1 release by using various mutated and chimaeric receptors transiently expressed in COS cells. Pulse-chase experiments revealed that (1) sCR1 is produced by a proteolytic process, (2) the cleavage site lies within the C-terminus of CR1 transmembrane domain, (3) the proteolytic process involves a fully glycosylated CR1 form and (4) this process takes place in late secretory vesicles or at the plasma membrane. PMID- 9405293 TI - Evidence for existence of tissue-specific regulation of the mammalian pyruvate dehydrogenase complex. AB - Tissue distribution and kinetic parameters for the four isoenzymes of pyruvate dehydrogenase kinase (PDK1, PDK2, PDK3 and PDK4) identified thus far in mammals were analysed. It appeared that expression of these isoenzymes occurs in a tissue specific manner. The mRNA for isoenzyme PDK1 was found almost exclusively in rat heart. The mRNA for PDK3 was most abundantly expressed in rat testis. The message for PDK2 was present in all tissues tested but the level was low in spleen and lung. The mRNA for PDK4 was predominantly expressed in skeletal muscle and heart. The specific activities of the isoenzymes varied 25-fold, from 50nmol/min per mg for PDK2 to 1250nmol/min per mg for PDK3. Apparent Ki values of the isoenzymes for the synthetic analogue of pyruvate, dichloroacetate, varied 40-fold, from 0.2 mM for PDK2 to 8 mM for PDK3. The isoenzymes were also different with respect to their ability to respond to NADH and NADH plus acetyl-CoA. NADH alone stimulated the activities of PDK1 and PDK2 by 20 and 30% respectively. NADH plus acetyl-CoA activated these isoenzymes nearly 200 and 300%. Under comparable conditions, isoenzyme PDK3 was almost completely unresponsive to NADH, and NADH plus acetyl CoA caused inhibition rather than activation. Isoenzyme PDK4 was activated almost 2-fold by NADH, but NADH plus acetyl-CoA did not activate above the level seen with NADH alone. These results provide the first evidence that the unique tissue distribution and kinetic characteristics of the isoenzymes of PDK are among the major factors responsible for tissue-specific regulation of the pyruvate dehydrogenase complex activity. PMID- 9405294 TI - Starvation and diabetes increase the amount of pyruvate dehydrogenase kinase isoenzyme 4 in rat heart. AB - This study investigated whether conditions known to alter the activity and phosphorylation state of the pyruvate dehydrogenase complex have specific effects on the levels of isoenzymes of pyruvate dehydrogenase kinase (PDK) in rat heart. Immunoblot analysis revealed a remarkable increase in the amount of PDK4 in the hearts of rats that had been starved or rendered diabetic with streptozotocin. Re feeding of starved rats and insulin treatment of diabetic rats very effectively reversed the increase in PDK4 protein and restored PDK enzyme activity to levels of chow-fed control rats. Starvation and diabetes also markedly increased the abundance of PDK4 mRNA, and re-feeding and insulin treatment reduced levels of the message to that of controls. In contrast with the findings for PDK4, little or no changes in the amounts of PDK1 and PDK2 protein and the abundance of their messages occurred in response to starvation and diabetes. The observed shift in the relative abundance of PDK isoenzymes probably explains previous studies of the effects of starvation and diabetes on heart PDK activity. The results indicate that control of the amount of PDK4 is important in long-term regulation of the activity of the pyruvate dehydrogenase complex in rat heart. PMID- 9405296 TI - Genes that cause aberrant cell morphology by overexpression in fission yeast: a role of a small GTP-binding protein Rho2 in cell morphogenesis. AB - To identify the genes involved in cell morphogenesis in Schizosaccharomyces pombe, we screened for the genes that cause aberrant cell morphology by overexpression. The isolated genes were classified on the basis of morphology conferred. One of the genes causing a rounded morphology was identified as the rho2+ gene encoding a small GTP-binding protein. The overexpression of rho2+ resulted in a randomized distribution of cortical F-actin and formation of a thick cell wall. Analyses using cdc mutants suggested that the overexpression of rho2+ prevents the establishment of growth polarity in G1. The rho2+ gene was not essential, but among cells deleted for rho2+, those with an irregular shape were observed. The disruptant also showed a defect in cell wall integrity. An HA-Rho2 expressed in the cell was suggested to be present as a membrane-bound form by a cell fractionation experiment. A GFP-Rho2 was localized at the growing end(s) of the cell and the septation site. The localization of GFP-Rho2 during interphase was partially dependent on sts5+. These results indicate that Rho2 is involved in cell morphogenesis, control of cell wall integrity, control of growth polarity, and maintenance of growth direction. Analysis of functional overlapping between Rho2 and Rho1 revealed that their functions are distinct from each other, with partial overlapping. PMID- 9405297 TI - Aminopeptidase B: a processing enzyme secreted and associated with the plasma membrane of rat pheochromocytoma (PC12) cells. AB - Aminopeptidase B (Ap-B) is a Zn2+-dependent exopeptidase which selectively removes Arg and/or Lys residues from the N terminus of several peptide substrates. Isolated and characterized from rat testes, this ubiquitous enzyme may participate in the final stages of precursor processing mechanisms. To test this hypothesis, we have investigated the secretion and subcellular localization of this enzyme in a rat cell line of pheochromocytoma (PC12 cells). By using a combination of biochemical and immunocytochemical methods, the following observations were made: (i) the level of aminopeptidase B detectable in the cell culture medium increased with time; (ii) 8-bromo-adenosine 3'-5'-cyclic monophosphate and the Ca2+ ionophore A23187 both stimulated enzyme liberation in the culture medium; (iii) brefeldin A, an inhibitor of vesicular transport from the endoplasmic reticulum to the Golgi apparatus, decreased enzyme secretion in a time-dependent manner; (iv) whereas nocodazole, a microtubule depolymerizing agent, inhibited enzyme secretion, cytochalasin D, a microfilament disruption agent, had no effect on released aminopeptidase B level; (v) immunofluorescence demonstrated the presence of aminopeptidase B in the Golgi apparatus; (vi) immunofluorescence, electron microscopy and tests of enzyme activity on intact cells showed an association of the peptidase with the external face of the plasma membrane. Together these data strongly argued in favour of the enzyme secretion by PC12 cells. It is concluded that aminopeptidase B may participate in processing events occurring either during its intracellular transport along the secretory pathway or at the plasma membrane level, or both. PMID- 9405295 TI - Subcellular co-localization and potential interaction of glucuronosyltransferases with nascent proteochondroitin sulphate at Golgi sites of chondroitin synthesis. AB - Microsomal membranes from chick embryo epiphyseal cartilage were fractionated by equilibrium sucrose-density-gradient centrifugation and assayed for GlcA (glucuronic acid) transferase I (the enzyme that transfers GlcA from UDP-GlcA to Gal-Gal-Xyl of proteochondroitin linkage region), for comparison with GlcA transferase II (the GlcA transferase of chondroitin polymerization). Gal(beta1 3)Galbeta1-methyl (disaccharide) and GalNAc(beta1-4)GlcA(beta1-3)GalNAc(beta1-4) GlcA(beta1-3)GalNAc(pentasaccharide) were used respectively as acceptors of [14C]GlcA from UDP-[14C]GlcA. Distributions of the two GlcA transferase activities in the sucrose-density-gradient fractions were compared with each other and with the previously reported distribution of the activities of Gal transferases (UDP-Gal to ovalbumin, and to xylose of the proteochondroitin linkage region) and GalNAc (N-acetylgalactosamine) transferase II of chondroitin polymerization. The linkage-region GlcA transferase I had a dual Golgi distribution similar to that of chondroitin-polymerizing GlcA transferase II and distinctly different from the distribution of linkage-region Gal transferases I and II, which were found exclusively in the heavier fractions. Solubilized GlcA transferase I was partly purified by sequential use of Q-Sepharose, heparin Sepharose and wheatgerm agglutinin-agarose and was accompanied at each step by some of the GlcA transferase II activity. Both GlcA transferase I and II bound to the Q-Sepharose as though they were highly anionic. However, treatment with chondroitin ABC lyase eliminated the binding while markedly decreasing enzyme stability. The enzyme activities could not be reconstituted by adding chondroitin or chondroitin pentasaccharide to the chondroitin ABC lyase-treated enzymes. Incubation of the partly purified enzymes with both UDP-GlcA and UDP-GalNAc resulted in a 40-fold greater incorporation than with just one sugar nucleotide, indicating the presence of bound, nascent proteochondroitin serving as the acceptor for chondroitin polymerization. These results, together with the membrane co-localization, indicate that GlcA transferase I and GlcA transferase II occur closely together with nascent proteochondroitin at the site of synthesis and that this complex with the nascent proteochondroitin stabilizes both enzymes during purification. PMID- 9405298 TI - Differential effects of temperature on specific and nonspecific immune defences in fish. AB - The susceptibility of fish to disease is partly dependent on their environment, in particular on water temperature. It is generally accepted that lower temperatures adversely affect specific immune responses mediated by T helper cells. The probable mechanisms involved in such suppression in teleost fish are reviewed. Furthermore, the effects of temperature on nonspecific defences, such as phagocytosis and cytotoxicity, are described and total immune competence in teleosts at low environmental temperatures is discussed. PMID- 9405299 TI - Specific behavioural responses triggered by identified mechanosensory receptor cells in the apical field of the giant rotifer Asplanchna sieboldi. AB - The giant rotifer Asplanchna sieboldi swims by the propulsive effect of thousands of cilia arrayed in clusters around the apical field, which has several mechanosensory structures (sensilla) located at defined positions. Males and females differ in both their patterns of behaviour and their sensory receptor equipment. Unstimulated males swim straight with occasional spontaneous changes in direction until they hit an obstacle with their apical field. Depending on the direction and the strength of the mechanical interference, the animals show different behavioural responses. To analyse the effect of excitation of the apical mechanosensitive sensilla on these responses, males were held on microcapillaries, and the sensitivity of individual sensilla was assayed using micromanipulator-mediated mechanical stimulation. Stimulation of each of the four different types of sensillum triggered a specific and well-defined initial behavioural response. Individual animals behaved identically with respect to the receptor specificity of the responses. The behaviour of free-swimming males upon contact with obstacles or females is discussed on the basis of these results. PMID- 9405300 TI - Suppression of tubulin tyrosine ligase during tumor growth. AB - The C terminus of the tubulin alpha-subunit of most eukaryotic cells undergoes a cycle of tyrosination and detyrosination using two specific enzymes, a tubulin tyrosine ligase (TTL) and a tubulin carboxypeptidase. Although this enzyme cycle is conserved in evolution and exhibits rapid turnover, the meaning of this modification has remained elusive. We have isolated several NIH-3T3 derived clonal cell lines that lack TTL (TTL-). TTL- cells contain a unique tubulin isotype (delta2-tubulin) that can be detected with specific antibodies. When injected into nude mice, both TTL- cells and TTL- cells stably transfected with TTL cDNA form sarcomas. But in tumors formed from TTL rescued cells, TTL is systematically lost during tumor growth. A strong selection process has thus acted during tumor growth to suppress TTL activity. In accord with this result, we find suppression of TTL activity in the majority of human tumors assayed with delta2-tubulin antibody. We conclude there is a widespread loss of TTL activity during tumor growth in situ, suggesting that TTL activity may play a role in tumor cell regulation. PMID- 9405301 TI - Effects of nonylphenol and 17 beta-oestradiol on vitellogenin synthesis, testicular structure and cytology in male eelpout Zoarces viviparus. AB - Nonylphenol has been found to be oestrogenic in fish and may influence the reproductive system of male fish. In the present study, the effects of low (10 microg g-1 week-1) and high (100 microg g-1 week-1) doses of nonylphenol and of 17 beta-oestradiol on the synthesis of vitellogenin and on testicular structure and cytology were investigated in male eelpout Zoarces viviparus during active spermatogenesis (May) and late spermatogenesis (June). Twenty-five days after injection, a significant dose-dependent increase in the plasma vitellogenin concentration, measured by enzyme-linked immunosorbent assay, was observed in the treated groups. A highly significant reduction in the gonadosomatic index was observed concomitant with the increase in the plasma vitellogenin concentration. Macroscopically, milt was observed to be present in the control fish, but was sparse or absent in the treated fish. Histological examination using light microscopy revealed severe effects of nonylphenol as well as of oestradiol treatment on testicular structure. Control fish had seminiferous lobules containing spermatogenic cysts and only a few spermatozoa (May) or had the walls of their seminiferous lobules lined with cuboidal Sertoli cells (June). In the treated fish, the seminiferous lobules were degenerated (May) or were filled with numerous spermatozoa and the Sertoli cells appeared very squamous (June). Electron microscopy revealed greater numbers of phagocytosed spermatozoa in these Sertoli cells. In rats, -glutamyl transpeptidase (-GTP) has been used as a specific marker of Sertoli cell function. In the present study, both nonylphenol and 17 beta-oestradiol treatment resulted in a reduction in the activity of this enzyme. The study provides evidence that nonylphenol is oestrogenic, as indicated by the large increase in vitellogenin synthesis, and that both nonylphenol and oestradiol have marked effects on the testicular structure and cytology of germ cells and Sertoli cells of male Z. viviparus. PMID- 9405302 TI - Messenger ribonucleoprotein complexes containing human ELAV proteins: interactions with cytoskeleton and translational apparatus. AB - Mammalian ELAV proteins bind to polyadenylated messenger RNAs and have specificity for AU-rich sequences. Preferred binding sites in vitro include the AUUUA pentamer and related sequences present in the 3' untranslated regions of many growth regulatory mRNAs. Human ELAV (hELAV) proteins have been implicated in post-transcriptional regulation of gene expression by their effects on the stability and translatability of growth regulatory mRNAs. We have examined the intracellular localization of ELAV proteins in neurons and in tumor cells of neuronal origin using indirect immunofluorescence, confocal microscopy and biochemical separation. Mammalian neuronal ELAV proteins are found predominantly in the cytoplasm of cells in mRNP complexes termed alpha complexes which, when associated with polysomes, form large and high density ss complexes, as assayed by glycerol and accudenz gradients, respectively. Puromycin, cytochalasin or EDTA treatments disrupt beta complexes causing the release of alpha complexes, which then appear, by confocal microscopy, as large hELAV mRNP granules associated with microtubules. Association of partially purified hELAV mRNP alpha complexes with microtubules was confirmed by in vitro reconstitution assays. Furthermore, colchicine treatment of cells suggested that association of hELAV mRNP alpha complexes with microtubules is also necessary for the formation of ss complexes. Our data suggest a model in which a subset of mRNAs is associated with microtubules as ELAV mRNP particles (alpha complexes) which, in turn, associate with polysomes to form a translational apparatus (beta complex) that is, through polysomes, associated with the microfilament cytoskeletal network. hELAV proteins in these mRNP granules may affect post-transcriptional regulation of gene expression via the intracellular transport, localization and/or translation of growth regulatory mRNAs. PMID- 9405303 TI - A peritracheal neuropeptide system in insects: release of myomodulin-like peptides at ecdysis. AB - We identified of a set of neuropeptide-expressing cells sited along the respiratory system of Drosophila melanogaster using an antibody to the molluscan neuropeptide myomodulin. The number and positions of these 'peritracheal' myomodulin (PM) cells were reminiscent of the epitracheal Inka cells in the moth Manduca sexta. These Inka cells release the peptide ecdysis-triggering hormone, which helps elicit ecdysial behavior at the molt, and we show that they are also recognized by the myomodulin (MM) antibody. In both D. melanogaster and M. sexta, the PM and Inka cells are the only MM-positive cells outside the central nervous system. In both insects, MM immunoreactivity disappears at the end of the molt. In D. melanogaster, we have monitored the PM cells throughout development using two enhancer trap lines; the PM cells persist throughout development, but at larval, pupal and adult ecdyses, they display a loss of MM immunoreactivity. This transient loss occurs at a predictable time, just prior to ecdysis. In contrast, MM-positive neurons in the central nervous system do not show these changes. The PM cells also reveal a concomitant loss of immunostaining for an enzyme contained in secretory granules. The results are consistent with the hypothesis that the PM cells release MM-like peptides just prior to each ecdysis. In addition, we demonstrate that peritracheal cells of five widely divergent insect orders show a myomodulin phenotype. The peritracheal cell size, morphology, numbers and distribution vary in these different orders. These data suggest that peritracheal cells release MM-like peptides as part of a conserved feature of the endocrine regulation of insect ecdysis. PMID- 9405304 TI - EGF stimulates an increase in actin nucleation and filament number at the leading edge of the lamellipod in mammary adenocarcinoma cells. AB - Stimulation of metastatic MTLn3 cells with EGF causes the rapid extension of lamellipods, which contain a zone of F-actin at the leading edge. In order to establish the mechanism for accumulation of F-actin at the leading edge and its relationship to lamellipod extension in response to EGF, we have studied the kinetics and location of EGF-induced actin nucleation activity in MTLn3 cells and characterized the actin dynamics at the leading edge by measuring the changes at the pointed and barbed ends of actin filaments upon EGF stimulation of MTLn3 cells. The major result of this study is that stimulation of MTLn3 cells with EGF causes a transient increase in actin nucleation activity resulting from the appearance of free barbed ends very close to the leading edge of extending lamellipods. In addition, cytochalasin D causes a significant decrease in the total F-actin content in EGF-stimulated cells, indicating that both actin polymerization and depolymerization are stimulated by EGF. Pointed end incorporation of rhodamine-labeled actin by the EGF stimulated cells is 2.12+/ 0.47 times higher than that of control cells. Since EGF stimulation causes an increase in both barbed and pointed end incorporation of rhodamine-labeled actin in the same location, the EGF-stimulated nucleation sites are more likely due either to severing of pre-existing filaments or de novo nucleation of filaments at the leading edge thereby creating new barbed and pointed ends. The timing and location of EGF-induced actin nucleation activity in MTLn3 cells can account for the observed accumulation of F-actin at the leading edge and demonstrate that this F-actin rich zone is the primary actin polymerization zone after stimulation. PMID- 9405305 TI - Energized Ca2+ transport by hepatopancreatic basolateral plasma membranes of Homarus americanus. AB - Ca2+ transport by hepatopancreatic basolateral membrane vesicles of Atlantic lobster (Homarus americanus) occurred by at least two independent processes: (1) an ATP-dependent carrier transport system, and (2) a Na+-gradient-dependent carrier mechanism. The sensitivity of ATP-dependent Ca2+ transport to vanadate indicated that it was probably due to a P-type ATPase. This system exhibited an extremely high apparent affinity for Ca2+ (Kt=65.28+/-14.39 nmol l-1; Jmax=1. 07+/-0.06 pmol microg-1 protein 8 s-1). The Na+-gradient-dependent carrier transport system exhibited the properties of a Ca2+/Na+ antiporter capable of exchanging external Ca2+ with intravesicular Na+ or Li+. Kinetic analysis of the Na+-dependence of the antiport indicated that at least three Na+ were exchanged with each Ca2+ (n=2. 91+/-0.22). When Li+ replaced Na+ in exchange for 45Ca2+, the apparent affinity for Ca2+ influx was not significantly affected (with Na+, Kt=14.57+/-5.02 micromol l-1; with Li+, Kt=20.17+/-6.99 micromol l-1), but the maximal Ca2+ transport velocity was reduced by a factor of three (with Na+, Jmax=2.72+/-0.23 pmol microg-1 protein 8 s-1; with Li+, Jmax=1.03+/-0.10 pmol microg-1 protein 8 s-1). It is concluded that Ca2+ leaves hepatopancreatic epithelial cells across the basolateral membrane by way of a high-affinity, vanadate-sensitive Ca2+-ATPase and by way of a low-affinity Ca2+/Na+ antiporter with an apparent 3:1 exchange stoichiometry. The roles of these transporters in Ca2+ balance during the molt cycle are discussed. PMID- 9405306 TI - Drosophila fascin mutants are rescued by overexpression of the villin-like protein, quail. AB - Actin bundle assembly in specialized structures such as microvilli on intestinal epithelia and Drosophila bristles requires two actin bundling proteins. In these systems, the distinct biochemical properties and temporal localization of actin bundling proteins suggest that these proteins are not redundant. During Drosophila oogenesis, the formation of cytoplasmic actin bundles in nurse cells requires two actin bundling proteins, fascin encoded by the singed gene and a villin-like protein encoded by the quail gene. singed and quail mutations are fully recessive and each mutation disrupts nurse cell cytoplasmic actin bundle formation. We used P-element mediated germline transformation to overexpress quail in singed mutants and test whether these proteins have redundant functions in vivo. Overexpression of quail protein in a sterile singed background restores actin bundle formation in egg chambers. The degree of rescue by quail depends on the level of quail protein overexpression, as well as residual levels of fascin function. In nurse cells that contain excess quail but no fascin, the cytoplasmic actin network initially appears wild type but then becomes disorganized in the final stages of nurse cell cytoplasm transport. The ability of quail overexpression to compensate for the absence of fascin demonstrates that fascin is partially redundant with quail in the Drosophila germline. Quail appears to function as a bundle initiator while fascin provides bundle organization. PMID- 9405307 TI - Different functions of different eye types in the spider Cupiennius salei. AB - The Central American hunting spider Cupiennius salei Keys relies mainly on its mechanosensory systems during prey-catching and mating behaviour. The behavioural relevance of its eight eyes has not been studied before, although their optics and sensitivity suggest highly developed visual capabilities. The visual system was examined in a twofold simultaneous-choice experiment. Two targets were presented at a distance of 2 m from the animals, and their walking paths towards the targets were monitored. Spiders showed no preference when choosing between two identical targets, but when choosing between two different targets they strongly preferred a vertical bar to a sloping bar or a V-shaped target. By covering all eyes except the anterior median or posterior median eyes, it could be shown that the spiders were able to detect the targets using any of the eyes. Discrimination between different targets was only possible with the anterior median eyes uncovered, although the visual fields of the anterior median and posterior median eyes overlap completely. It seems most likely that the animals separate visual information in the periphery and therefore that the eyes have different functions. The posterior median eyes support a target-detecting mechanism and the anterior median eyes a target-discrimination mechanism. PMID- 9405308 TI - Active nuclear pore complexes in Chironomus: visualization of transporter configurations related to mRNP export. AB - The Nuclear Pore Complex (NPC) regulates nucleocytoplasmic transport by providing small channels for passive diffusion and multiple docking surfaces that lead to a central translocation channel for active transport. In this study we have investigated by high resolution scanning and transmission electron microscopy the dynamics of NPC structure in salivary gland nuclei from Chironomus during Balbiani ring (BR) mRNP translocation, and present evidence of rearrangement of the transporter related to mRNP export. Analysis of the individual NPC components verified a strong evolutionary conservation of NPC structure between vertebrates and invertebrates. The transporter is an integral part of the NPC and is composed of a central short double cylinder that is retained within the inner spoke ring, and two peripheral globular assemblies which are tethered to the cytoplasmic and nucleoplasmic coaxial rings by eight conserved internal ring filaments. Distinct stages of BR mRNP nuclear export through the individual NPC components were directly visualized and placed in a linear transport sequence. The BR mRNP first binds to the NPC basket, which forms an expanded distal basket ring. In this communication we present stages of BR mRNP transport through the nucleoplasmic, central and cytoplasmic transporter subunits, which change their conformation during mRNP translocation, and the emergence of mRNP into the cytoplasm. We propose that the reorganization of the basket may be driven, in part, by an active translocation process at the transporter. Furthermore, the images provide dramatic evidence that the transporter functions as a central translocation channel with transiently open discrete gates in its globular assemblies. A model of NPC transporter reorganization accompanied with mRNP translocation is discussed. PMID- 9405309 TI - Freezing survival by isolated Malpighian tubules of the New Zealand alpine weta Hemideina maori. AB - The ability of isolated Malpighian tubules from a freeze-tolerant insect, the New Zealand alpine weta (Hemideina maori), to withstand freezing was assessed by measuring post-freeze membrane potentials and rates of fluid secretion. The hemolymph of cold-acclimated Hemideina maori was found to contain relatively high concentrations of the cryoprotectants trehalose (>300 mmol l-1) and proline (41 mmol l-1). Survival of isolated Malpighian tubules was correspondingly high when a high concentration of trehalose was present in the bathing saline. Tubules allowed to recover for 20 min from a 1 h freeze to -5 degrees C in saline containing 400 mmol l-1 trehalose had a basolateral membrane potential of -53 mV compared with a potential of -63 mV in tubules not exposed to a freeze/thaw cycle. Fluid secretion in tubules that had experienced a freeze/thaw cycle in saline containing 400 mmol l-1 trehalose was 9.9+/-2.6 nl h-1 compared with 18.7+/-5.0 nl h-1 (means +/- s.e.m., N=18) in tubules that had not been frozen. Tubules frozen in saline containing a lower concentration of trehalose (200 mmol l-1) or in glucose (400 mmol l-1) showed a similar ability to survive freezing to -5 degrees C. In contrast, freezing for 1 h at -5 degrees C in saline containing 400 mmol l-1 sucrose produced a 57 % decrease in membrane potential and an 88 % decrease in secretion rate. Tubules held in saline lacking high concentrations of sugars showed no survival after freezing to -5 degrees C for 1 h. When frozen to 15 degrees C, tubules appeared to survive best in saline with the highest trehalose concentration (400 mmol l-1). Freezing damage was not simply the result of exposure to cold, since tubules chilled (unfrozen) to -5 degrees C for 1 h were not compromised even when the bathing saline lacked a high sugar concentration. Exposure of tubules to a combination of low temperature and high osmolality mimicked damage caused by actual freezing: the membrane potential showed a 60 % recovery when the test was performed in saline containing trehalose, but showed no recovery in saline containing sucrose. PMID- 9405310 TI - Agonistic monoclonal antibodies against the Met receptor dissect the biological responses to HGF. AB - Hepatocyte growth factor, also known as scatter factor, is a pleiotropic cytokine, which stimulates cell motility, invasion, proliferation, survival and morphogenesis, and induces the expression of specific genes by activating its receptor tyrosine kinase. In this work we have isolated, characterized and used as agonists two monoclonal antibodies (mAbs) directed against the extracellular domain of HGF receptor to investigate the requirements for receptor activation and for the different biological responses. The two mAbs display similar affinities, react with epitopes different from the hepatocyte growth factor binding site, and behave as either full or partial agonists. The full agonist mAb (DO-24) triggers all the biological effects elicited by hepatocyte growth factor, namely motility, proliferation, cell survival, invasion, tubulogenesis and angiogenesis. The partial agonist mAb (DN-30) induces only motility. Only the full agonist mAb is able to induce and sustain the expression of urokinase-type plasminogen activator receptor for prolonged periods of time, while both mAbs up regulate the constitutive expression of urokinase-type plasminogen activator. Both mAbs activate receptor phosphorylation, which, being strictly dependent on mAb bivalence, requires receptor dimerization. Since simple receptor dimerization is not sufficient to trigger full biological responses, we propose that the region on the ss chain of the receptor recognized by the full agonist mAb is crucial for optimal receptor activation. PMID- 9405311 TI - Gas exchange during hovering flight in a nectar-feeding bat Glossophaga soricina. AB - Glossophagine nectar-feeding bats exploit flowers while hovering in front of them. Aerodynamic theory predicts that power output for hovering flight in Glossophaga soricina is 2.6 times higher than during horizontal flight. We tested this prediction by measuring rates of gas exchange during hover-feeding. Five individuals of Glossophaga soricina (mean mass 11.7 g) were trained to feed from a nectar dispenser designed as a flow-through respirometry mask. Single hover feeding events lasted for up to 4.5 s. Measured rates of gas exchange varied as a function of hovering duration. O2 and CO2 during short hovering events (up to 1 s) were 20.5+/-6.7 ml g-1 h-1 (N=55) and 21.6+/-5.6 ml g-1 h-1 (N=39) (means +/- S.D.), respectively. These values are in the range of a previous estimate of the metabolic power input for level forward flight (23.8 ml O2 g-1 h-1). However, during hovering events lasting longer than 3 s, oxygen uptake was only 16.7+/-3.5 ml g-1 h-1 (N=73), which is only 70 % of the value expected for forward flight. Thus, bats reduced their rate of oxygen uptake during longer periods of hovering compared with level forward flight. This result is in contrast to the predicted hovering cost derived from aerodynamic theory. The exact metabolic power input during hovering remains uncertain. During longer hovering events, bats were probably not in respiratory steady state, as indicated by the deviation of the respiratory exchange ratio from the expected value of 1 (oxidization of nectar sugar) to the measured value of 0.8. PMID- 9405312 TI - Modelling quantitative structure-activity relationships between animal behaviour and environmental signal molecules. AB - Quantitative structure-activity relationships (QSARs) between the physicochemical properties of environmental signal molecules and animal behaviour have been determined. Past work has shown that oyster and barnacle larval settlement and mud crab abdominal pumping (for larval dispersal) are stimulated by small peptide cues. In all the peptides examined that were active at ecologically relevant concentrations, arginine or lysine was found at the carboxy terminus, but the amino acids found at preceding positions were highly variable. We used the multivariate partial least squares algorithm to relate composite properties for the hydrophilicity, size and charge of each amino acid and the sequence position to oyster, barnacle and crab behaviour patterns. From the information in these QSAR models, the apparent variability in amino acid sequences eliciting behavioural responses was explained in each case, and more potent peptide analogues are hypothesized on the basis of untested amino acid sequences. Remarkably, these peptide signals are all structurally related to the carboxy terminal sequence of mammalian C5a anaphylatoxin, a potent white blood cell chemoattractant. Even more striking is the fact that these different animal species should rely on apparently similar environmental signal molecules when residing within a common habitat (southeastern US estuaries). Through the physicochemical properties of amino acids, the current QSAR models clearly differentiate between the optimal sequences for eliciting oyster, barnacle and mud crab behaviour. Thus, QSARs provide a novel and powerful method not only for relating the physicochemical properties of molecules to animal behaviour but also for differentiating responses to chemicals by individuals of different species. PMID- 9405314 TI - Persistent desensitisation of the beta 2 adrenoceptors expressed by cultured equine sweat gland epithelial cells. AB - Adrenaline, forskolin and ATP all evoked accumulation of cyclic AMP in equine sweat gland epithelial cells, although the response to adrenaline was more transient than that to forskolin and ATP. Cells preincubated in adrenaline (10 micromol l-1, 32 min) showed essentially complete, homologous desensitisation, and this phenomenon reversed slowly (half-time 6.3+/-0.9 h). After 10 min of recovery from preincubation in adrenaline, isobutylmethylxanthine (IBMX, 5 mmol l 1) had no effect upon the desensitisation and the cells showed no loss of sensitivity to ATP and forskolin. After 10 h, however, the persistent desensitisation was partially reversed by IBMX and the cells showed reduced responses to ATP and forskolin. Increased phosphodiesterase activity may thus contribute to the persistent desensitisation. Experiments using forskolin preincubated (100 micromol l-1, 32 min) cells suggested that increased cytosolic cyclic AMP levels did not underlie the initial loss of sensitivity to adrenaline but that this second messenger may initiate the series of events leading to the generalised loss of sensitivity seen after 10 h. PMID- 9405313 TI - Truncations of the C-terminal cytoplasmic domain of MG160, a medial Golgi sialoglycoprotein, result in its partial transport to the plasma membrane and filopodia. AB - MG160, a type I cysteine-rich membrane sialoglycoprotein residing in the medial cisternae of the rat Golgi apparatus, is highly homologous to CFR, a fibroblast growth factor receptor, and ESL-1, an E-selectin ligand located at the cell surface of mouse myeloid cells and recently detected in the Golgi apparatus as well. The mechanism for the transport of MG160 from the Golgi apparatus to the cell surface is unknown. In this study we found that differential processing of the carboxy-terminal cytoplasmic domain (CD), consisting of amino acids Arg1159 Ile Thr Lys Arg Val Thr Arg Glu Leu Lys Asp Arg1171, resulted in the partial transport of the protein to the plasma membrane and filopodia. In Chinese hamster ovary cells (CHO), stably transfected with the entire cDNA encoding MG160, the protein was localized in the Golgi apparatus. However, when the terminal Arg1171 or up to nine distal amino acids were deleted, the protein was distributed to the plasma membrane and filopodia as well as the Golgi apparatus. This report shows that the CD of an endogenous type I Golgi protein is important for its efficient retention and identifies a unique residue preference in this process. Cleavage within the CD of MG160 may constitute a regulatory mechanism for the partial export of the protein from the Golgi apparatus to the plasma membrane and filopodia. PMID- 9405315 TI - Annexin VI defines an apical endocytic compartment in rat liver hepatocytes. AB - Annexin VI has been demonstrated previously to be a marker for hepatic endosomes. By western blotting with an affinity purified anti-annexin VI antibody it was shown that annexin VI was present in the three morphologically and functionally different endosomal fractions from rat liver. We have quantified the gold-labeled endosomes by immunoelectron microscopy in ultrathin Lowicryl sections of rat liver and now demonstrate that 80% of the total labeling with anti-annexin VI was associated with endocytic structures surrounding the bile canaliculus, the apical domain of hepatocytes, whereas only 20% was found in the subsinusoidal endosomes. In double immuno-gold labeling experiments 80% of the Rab5 positive apical endosomes were also labeled with anti-annexin VI antibodies. However, there was no significant colocalization with antibodies to the polymeric immunoglobulin receptor. Finally, we demonstrate that 50% of endosomes containing internalized gold-labeled transferrin were double labeled with anti-annexin VI antibodies. Thus, annexin VI becomes the first known structural protein at the apical 'early' endocytic compartment of the hepatocyte that may be involved in the receptor recycling and transport to late endocytic/lysosomal compartment pathways. PMID- 9405316 TI - Cardiovascular responses in vivo to angiotensin II and the peptide antagonist saralasin in rainbow trout Oncorhynchus mykiss. AB - The effects of [Asn1,Val5]-angiotensin II (AngII) and [Sar1,Val5, Ala8] angiotensin II (saralasin) on dorsal aortic blood pressure, pulse pressure and heart rate were examined in rainbow trout in vivo. AngII when administered as a single dose of 25 microg kg-1 induced a biphasic response in blood pressure, with a significant hypertensive response during the initial 10 min, followed by a significant hypotension of 70-75 % compared with the initial blood pressure after 50 min and continuing until approximately 80 min post-injection. The co administration of AngII (25 microg kg-1) and saralasin (50 microg kg-1) resulted in the same hypertensive response during the initial phase, but abolished the hypotensive effect of AngII. Heart rate was significantly increased in response to AngII, but the administration of AngII and saralasin together attenuated the increase by approximately 44 %. Stimulation of the endogenous renin-angiotensin system using a vasodilator, sodium nitroprusside, significantly increased drinking rate in rainbow trout fry, a response inhibited by saralasin, indicating a role for AngII-induced hypotension in drinking. For the first time, a decrease in blood pressure in response to AngII in vivo has been demonstrated in fish, and this is discussed in relation to homeostasis of blood pressure and a possible role in the control of drinking. PMID- 9405317 TI - An image correlation analysis of the distribution of clathrin associated adaptor protein (AP-2) at the plasma membrane. AB - Clathrin associated adaptor protein is involved in endocytosis at the plasma membrane (AP-2) and protein sorting at the Golgi membrane (AP-1). There is a great deal of information available on the structure, function and binding characteristics of AP-2, however, there is little quantitative data on the AP-2 distribution at the membrane. Image correlation spectroscopy is a technique which yields number counts from an autocorrelation analysis of intensity fluctuations within confocal microscopy images. Image correlation spectroscopy analysis of the indirect immunofluorescence from AP-2 at the plasma membrane of CV-1 cells shows that AP-2 is in a bimodal distribution consisting of large coated pit associated aggregates of approximately 60 AP-2 molecules, and smaller aggregates containing approximately 20 AP-2 molecules, which we propose are coated pit nucleation sites. Following hypertonic treatment 25% of the AP-2 molecules dissociate from the large AP-2 aggregates and form AP-2 dimers, leaving the remaining AP-2 as large aggregates with approximately 45 molecules. The smaller AP-2 aggregates completely dissociate forming AP-2 dimers. Dispersion of AP-2 with hypertonic treatment is not seen qualitatively because the number of large AP-2 aggregates is unchanged, the aggregates are just 25% smaller. Change in temperature from 37 degrees C to 4 degrees C has no affect on the number of AP-2 aggregates or the AP 2 distribution between the two populations. These data and estimates of the coated pit size suggest that coated pits cover approximately 0.9% of the cell membrane. Combination of image correlation spectroscopy analysis and measurements of the CV-1 cell surface area show that there are approximately 6x10(5) AP-2 molecules per CV-1 cell with approximately 2x10(5) AP-2 molecules within coated pit structures. PMID- 9405318 TI - Effects of incline on speed, acceleration, body posture and hindlimb kinematics in two species of lizard Callisaurus draconoides and Uma scoparia. AB - We examined the effects of incline on locomotor performance and kinematics in two closely related species of iguanian lizards that co-occur in sandy desert habitats. Callisaurus draconoides differs from Uma scoparia of equal snout-vent length by being less massive and having greater limb and tail lengths. We analyzed high-speed video tapes of lizards sprinting from a standstill on a sand covered racetrack which was level or inclined 30 degrees uphill. C. draconoides sprinted significantly faster than U. scoparia on both level and uphill sand surfaces, although U. scoparia is considered to be more specialized for sandy habitats. Initial accelerations (over the first 50 ms) did not differ significantly either between species or between inclines within species. Overall, the effects of incline were more pronounced for C. draconoides than for U. scoparia. For example, the incline caused a significant decrease in the maximum stride length of C. draconoides but not in that of U. scoparia. For C. draconoides, uphill stride durations were significantly shorter than on the level surface, and this partially compensated for the effects of shorter uphill stride lengths on velocity. C. draconoides ran bipedally more often than did U. scoparia on both the level and uphill surfaces. PMID- 9405319 TI - Disruption of Dictyostelium PI3K genes reduces [32P]phosphatidylinositol 3,4 bisphosphate and [32P]phosphatidylinositol trisphosphate levels, alters F-actin distribution and impairs pinocytosis. AB - To understand how phosphatidylinositol 3-kinase (PI3K) modulates cell structure and function, we examined the molecular and cellular defects of a Dictyostelium mutant strain (pik1(Delta)2(Delta)) missing two (DdPIK1 and 2) of three PI3K genes, which are homologues of the mammalian p110 subunit. Levels of [32P]phosphatidylinositol 3, 4 bisphosphate (PI(3,4)P2) and [32P]phosphatidylinositol trisphosphate (PIP3) were reduced in pik1(Delta)2(Delta), which had major defects in morphological and functional correlates of macropinocytosis. This was accompanied by dramatic deficits in a subset of F-actin-enriched structures such as circular ruffles, actin crowns and pseudopodia. Although pik1(Delta)2(Delta) were mobile, they failed to aggregate into streams. Therefore we conclude that PIK1 and 2, possibly through modulation of the levels of PIP3 and PI(3,4)P2, regulate the organization of actin filaments necessary for circular ruffling during macropinocytosis, the extension of pseudopodia and the aggregation of cells into streams, but not the regulation of cell motility. PMID- 9405320 TI - Adenosine and anoxia reduce N-methyl-D-aspartate receptor open probability in turtle cerebrocortex. AB - During normoxia, glutamate and the glutamate family of ion channels play a key role in mediating rapid excitatory synaptic transmission in the central nervous system. However, during hypoxia, intracellular [Ca2+] increases to neurotoxic levels, mediated largely by the N-methyl-D-aspartate (NMDA) subfamily of glutamate receptors. Adenosine has been shown to decrease the magnitude of the hypoxia-induced increase in [Ca2+]i in mammalian brain slices, delaying tissue injury. Turtle brain is remarkably tolerant of anoxia, maintaining a pre-anoxic [Ca2+]i while cerebral adenosine levels increase 12-fold. Employing cell-attached single-channel patch-clamp techniques, we studied the effect of adenosine (200 micromol l-1) and anoxia on NMDA receptor open probability (Popen) and current amplitude. After 60 min of anoxic perfusion, channel Popen decreased by 65 % (from 6.8+/-1.6 to 2.4+/-0.8 %) an effect that could also be achieved with a normoxic perfusion of 200 micromol l-1 adenosine (Popen decreased from 5.8+/-1.1 to 2.3+/-1.2 %). The inclusion of 10 micromol l-1 8-phenyltheophylline, an A1 receptor blocker, prevented the adenosine- and anoxia-induced decrease in Popen. Mean single-channel current amplitude remained at approximately 2.7+/-0.23 pA under all experimental conditions. To determine whether a change in the membrane potential could be part of the mechanism by which Popen decreases, membrane and threshold potential were measured following each experiment. Membrane potential did not change significantly under any condition, ranging from -76.8 to -80.6 mV. Therefore, during anoxia, NMDA receptors cannot be regulated by Mg2+ in a manner dependent on membrane potential. Threshold potentials did decrease significantly following 60 min of anoxic or adenosine perfusion (control -33.3+/-1.9 mV, anoxia -28.4+/-1.5 mV, adenosine -23.4+/-2.8 mV). We conclude that anoxia modulates NMDA receptor activity and that adenosine plays a key role in mediating this change. This is the first direct measurement of ion channel activity in anoxic turtle brain and demonstrates that ion channel regulation is part of the naturally evolved anoxic defence mechanism of this species. PMID- 9405321 TI - Lead poisoning--one approach to a problem that won't go away. AB - A reduction in sources of environmental lead exposure has resulted in substantial declines in mean blood lead concentrations of all age groups in the United States. However, some segments of the population continue to have unacceptable levels of lead exposure and elevated blood lead concentrations. In addition, virtually all residents of industrialized countries have bone lead stores that are several orders of magnitude greater than those of our preindustrial ancestors. Recent studies suggest that these skeletal lead stores adversely affect health and can contribute to reduced birth weights, aggressive behavior in children, and anemia, hypertension, and kidney disease in adults. Evidence is described that demonstrates that an increase in dietary calcium consumption can reduce lead absorption and toxicity from exogenous and endogenous lead exposure. A relatively inexpensive and effective way to reduce the substantial morbidity that will result from widespread lead exposure is by fortification of a variety of foods with low levels of calcium. This approach can complement other efforts to prevent lead exposure and reduce lead toxicity. PMID- 9405324 TI - Integration of human health and ecological risk assessment. PMID- 9405323 TI - Soil ingestion: a concern for acute toxicity in children. AB - Several soil ingestion studies have indicated that some children ingest substantial amounts of soil on given days. Although the EPA has assumed that 95% of children ingest 200 mg soil/day or less for exposure assessment purposes, some children have been observed to ingest up to 25-60 g soil during a single day. In light of the potential for children to ingest such large amounts of soil, an assessment was made of the possibility for soil pica episodes to result in acute intoxication from contaminant concentrations the EPA regards as representing conservative screening values (i.e., EPA soil screening levels and EPA Region III risk-based concentrations for residential soils). For a set of 13 chemicals included in the analysis, contaminant doses resulting from a one-time soil pica episode (5-50 g of soil ingested) were compared with acute dosages shown to produce toxicity in humans in clinical studies or case reports. For four of these chemicals, a soil pica episode was found to result in a contaminant dose approximating or exceeding the acute human lethal dose. For five of the remaining chemicals, the contaminant dose from a soil pica episode was well within the reported dose range in humans for toxicity other than lethality. Because both the exposure episodes and the toxicological response information are derived from observations in humans, these findings are regarded as particularly relevant for human health risk assessment. They suggest that, for some chemicals, ostensibly conservative soil criteria based on chronic exposure using current EPA methodology may not be protective of children during acute soil pica episodes. PMID- 9405322 TI - An alternative approach for investigating the carcinogenicity of indoor air pollution: pets as sentinels of environmental cancer risk. AB - Traditionally, the cancer risks associated with radon,environmental tobacco smoke (ETS), and similar indoor residential exposures have been evaluated through either laboratory experiments in rodents or epidemiology studies in people. Laboratory studies have the advantage of being controlled experiments, but their utility as estimators of human risk is limited by the uncertainties of extrapolating from rodents to people and from high doses to those typically experienced in the home. These experiments also subject animals to noxious exposures, causing suffering that may be considered cruel. Traditional epidemiology studies evaluate human risk directly, at the exposure levels present in residences; however, these studies are limited by their potential for misclassification, biased recall, and uncontrolled confounding. The long time intervals involved between exposure and disease (often 30 years or more) make accurate recall particularly problematic. In this paper we discuss the limitations of these traditional approaches, especially as they relate to residential studies of radon and ETS. The problems associated with the maximum tolerated dose in rodent bioassays and exposure misclassification in traditional epidemiology are particularly examined. A third approach that supplements the traditional approaches and overcomes some of their limitations is suggested. This approach, dubbed pet epidemiology, estimates residential cancer risk by examining the exposure experience of pet dogs with naturally occurring cancers. The history of pet epidemiology is reviewed and its strengths and limitations are examined. PMID- 9405325 TI - Plane pollution. PMID- 9405326 TI - Dietary lead intakes for mother/child pairs and relevance to pharmacokinetic models. AB - Blood and environmental samples, including a quarterly 6-day duplicate diet, for nine mother/child pairs from Eastern Europe have been monitored for 12 to >24 months with high precision stable lead isotope analysis to evaluate the changes that occur when the subjects moved from one environment (Eastern Europe) to another with different stable lead isotopes (Australia). The children were between 6 and 11 years of age and the mothers were between 29 and 37 years of age. These data were compared with an Australian control mother/child pair, aged 31 and 6 years, respectively. A rationale for undertaking this study of mother/child pairs was to evaluate if there were differences in the patterns and clearance rates of lead from blood in children compared with their mothers. Blood lead concentrations ranged from 2.1 to 3.9 microg/dl in the children and between 1.8 and 4.5 microg/dl in the mothers, but the mean of differences between each mother and her child did not differ significantly from zero. Duplicate diets contained from 2.4 to 31.8 microg Pb/kg diet; the mean+/- standard deviation was 5.5 +/- 2.1 microg Pb/kg and total daily dietary intakes ranged from 1.6 to 21.3 microg/day. Mean daily dietary intakes relative to body weight showed that the intake for children was approximately double that for the mothers (0.218 vs. 0. 113 microg Pb/kg body weight/day). The correlations between blood lead concentration and mean daily dietary intake either relative to body weight or total dietary intake did not reach statistical significance (p>0.05). Estimation of the lead coming from skeletal (endogenous) sources relative to the contribution from environmental (exogenous) sources ranges from 8 to 70% for the mothers and 12 to 66% for the children. The difference between mothers and children is not statistically significant (p = 0.28). The children do not appear to achieve the Australian lead isotopic profile at a faster rate than their mothers. These data provide evidence that the absorption or uptake of lead from dietary sources is similar in adult females and children of the age in this study. In spite of lower bone lead and faster bone remodeling and recycling in children compared with adult females, we see no differences between the mothers and their children in overall contribution of tissue lead to blood lead. Results from this study suggest that fractional absorption of ingested lead by children 6 11 years of age is comparable with absorption patterns observed among adult females in the 29-37-year-old age range. Because pharmacokinetic models apply a 40-50% absorption even for 7-year-old children, further investigations on fractional absorption of ingested lead by young children are warranted. Further investigations are especially needed in younger children than those who were subjects in the current study, particularly children in the 1-3-year-old age range. In addition, the effect of nutritional status and patterns of food intake on children's lead absorption require investigation, particularly given the increased prevalence of marginal nutritional status among low-income populations that are at increased risk of elevated blood lead levels. PMID- 9405327 TI - Fast-track cloning. PMID- 9405328 TI - Persisting learning deficits in rats after exposure to Pfiesteria piscicida. AB - Pfiesteria piscicida and other toxic Pfiesteria-like dinoflagellates have been implicated as a cause of fish kills in North Carolina estuaries and elsewhere. Accidental laboratory exposure of humans to P. piscicida has been reported to cause a complex syndrome including cognitive impairment. The current project was conducted to experimentally assess the possibility of cognitive effects of P. piscicida exposure in rats. Samples of water from aquaria in which P. piscicida zoospores were killing fish were frozen, a procedure that has been found to induce encystment. Thawed samples were injected into albino Sprague-Dawley rats. A significant learning impairment was documented in rats administered samples of P. piscicida that were recently frozen. Prolonged storage of Pfiesteria samples diminished the effect. No effect was seen in the recall of a previously learned task, but when the rats were called upon to learn a new task, the Pfiesteria treated animals showed a significant learning deficit. This effect persisted up to at least 10 weeks after a single injection of Pfiesteria. The Pfiesteria induced learning deficit did not seem to be associated with any obvious debilitation or health impairment of the exposed rats. Deficits in habituation of arousal and rearing behavior were detected using a functional observational battery. No Pfiesteria-induced effects on blood count and white cell differential or in a standard pathological screening of brain, liver, lung, kidney, and spleen tissue were seen at 2 months after exposure. These studies document a persistent learning impairment in rats after exposure to the dinoflagellate P.piscicida in otherwise physically well-appearing rats. This effect may partially model the symptoms of cognitive impairments that humans have shown after Pfiesteria exposure. PMID- 9405329 TI - Biological monitoring of organophosphorus pesticide exposure among children of agricultural workers in central Washington State. AB - Children up to 6 years of age who lived with pesticide applicators were monitored for increased risk of pesticide exposure: 48 pesticide applicator and 14 reference families were recruited from an agricultural region of Washington State in June 1995. A total of 160 spot urine samples were collected from 88 children, including repeated measures 3-7 days apart. Samples were assayed by gas chromatography flame photometric detector for dimethylphosphate metabolites. Dimethylthiophosphate (DMTP) was the dominant metabolite. DMTP levels were significantly higher in applicator children than in reference children (p = 0.015), with median concentrations of 0.021 and 0.005 microg/ml, respectively; maximum concentrations were 0.44 and 0.10 microg/ml, respectively. Percentages of detectable samples were 47% for applicator children and 27% for reference children. A marginally significant trend of increasing concentration was observed with decreasing age among applicator children (p = 0.060), and younger children within these families had significantly higher concentrations when compared to their older siblings (p = 0.040). Applicator children living less than 200 feet from an orchard were associated with higher frequency of detectable DMTP levels than nonproximal applicator children (p =0.036). These results indicate that applicator children experienced higher organophosphorus pesticide exposures than did reference children in the same community and that proximity to spraying is an important contributor to such exposures. Trends related to age suggest that child activity is an important variable for exposure. It is unlikely that any of the observed exposures posed a hazard of acute intoxication. This study points to the need for a more detailed understanding of pesticide exposure pathways for children of agricultural workers. PMID- 9405330 TI - Health advisories for consumers of Great Lakes sport fish: is the message being received? AB - Nationwide, 45 states issue health advisories for sport fish consumers. Chemical contaminants in some Great Lakes (GL) sport fish include compounds suspected of causing adverse reproductive and developmental effects. Although advisories to reduce consumption of contaminated fish, especially by women, have been issued by GL states (i.e., Illinois, Indiana, Michigan, Minnesota, New York, Ohio, Pennsylvania, and Wisconsin) since the mid-1970s, little is known about advisory awareness and GL sport fish consumption in the general population. To estimate the prevalence of GL sport fish consumption and health advisory awareness, we conducted a population-based telephone survey of 8,306 adult residents of the eight GL states. We gathered information concerning respondents' demographic characteristics, fish consumption during the preceding year, and sport fish consumption advisory awareness. The survey response rate was 69%. GL sport fish were eaten during the preceding year by 8.4% -95% confidence interval (CI), 7.6 9.2- of adults in the GL states, approximately 4.7 million persons. Women accounted for 43.9% (CI,39. 4-48.4) of consumers. Although 49.9% of GL sport fish consumers were aware of a health advisory, awareness varied significantly by sex: 58.2% (CI, 51.7-64.7) of males and 39.1% (CI, 32.6-45.6) of females were aware. Using logistic regression, we found awareness associated with male sex -odds ratio (OR) = 2.3; CI, 1.5-3.5), white race (OR = 4.2; CI, 1.9-9.1), college degree (OR = 3.1; CI, 1.3-7.6), and consuming >=24 GL sport fish meals/year (OR = 2.4; CI, 1.4-4.3). Only half of GL sport fish consumers reported awareness of a health advisory concerning eating GL sport fish. Awareness was especially low among women, suggesting the need of targeted risk communication programs for female consumers. PMID- 9405331 TI - Dioxinlike components in incinerator fly ash: a comparison between chemical analysis data and results from a cell culture bioassay. AB - Potent polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and dioxinlike polychlorinated biphenyls (PCBs) are among the most relevant toxic emissions from incinerators. Induction of cytochrome P450 1A1 catalyzed 7-ethoxyresorufin O-deethylase (EROD) activity in mammalian cell culture (EROD bioassay) is thought to be a selective and sensitive parameter used for the quantification of dioxinlike compounds. Fly ash extracts from municipal waste incinerators (MWI), a crematorium, wood combustors, and a noble metal recycling facility were analyzed in the EROD bioassay using rat hepatocytes in primary culture. Fractions containing 2,3,7,8-substituted PCDDs/PCDFs, dioxinlike PCBs, and 16 major polycyclic aromatic hydrocarbons (PAHs) were isolated from the extract and analyzed by gas chromatography-mass spectrometry (GC-MS) and by the EROD bioassay. It was found that with MWI samples the bioassay of the extract resulted in a two- to fivefold higher estimate of TCDD equivalents (TEQ) than the chemical analysis of PCDDs/PCDFs and PCBs. However, the outcome of both methods was significantly correlated, making the bioassay useful as a rough estimate for the sum of potent PCDDs/PCDFs and dioxinlike PCBs in extracts from MWI fly ash samples and in a fly ash sample from a crematorium. In noble metal recycling facility and wood combustor samples, higher amounts of PAHs were found, contributing to more pronounced differences between the results of both methods. The remaining unexplained inducing potency in fly ash samples probably results from additional dioxinlike components including certain PAHs not analyzed in this study. The hypothesis that emissions from MWI of hitherto unidentified dioxinlike compounds are higher by orders of magnitude than emissions of potent PCDDs/PCDFs and dioxinlike PCBs could not be confirmed. We found no indication for a marked synergistic interaction of dioxinlike fly ash components in the bioassay. PMID- 9405333 TI - Diagnostic strategies in Pneumocystis carinii pneumonia. AB - Pneumocystis carinii (P. carinii) remains a major pulmonary pathogen for the immunocompromised patient. In HIV infected patients, P. carinii represents the most commonly diagnosed cause of pneumonia. In the AIDS patient, empiric therapy based on clinical presentation has its proponents. However, this approach has been associated with a worse overall prognosis for the at risk patient. Because P. carinii can not be cultured, specific identification relies on examining respiratory specimens ranging from expectorated sputum to bronchoscopy with bronchoalveolar lavage (BAL). The low sensitivity of conventional stains has led to the search for antibodies to P. carinii and the use of immunofluorescent techniques. In addition, the polymerase chain reaction (PCR) is successfully being used in the diagnosis of P. carinii. Overall, these techniques allow the clinician to tailor the diagnostic testing for the individual patient. PMID- 9405334 TI - Ascorbate function and metabolism in the human erythrocyte. AB - Ascorbic acid, or vitamin C, is an important antioxidant in plasma, where it consumes oxygen free radicals and helps to preserve alpha-tocopherol (vitamin E) in lipoproteins. Erythrocytes, as the most plentiful cell in blood, help to preserve ascorbate in the blood plasma. In contrast to nucleated cells, which avidly concentrate ascorbate, the erythrocyte ascorbate concentration is the same as that in plasma. Erythrocytes nonetheless have a high capacity to regenerate the vitamin from its two electron-oxidized form, dehydroascorbic acid (DHA). DHA is rapidly taken up by these cells on the abundant glucose transport protein, GLUT1. Intracellular DHA is rapidly reduced to ascorbate by GSH in a direct chemical reaction, although enzyme-dependent mechanisms involving both glutaredoxin and thioredoxin reductase have also been demonstrated. Ascorbate, which carries a negative charge at physiologic pH, enters and leaves the cells slowly. Nonetheless, this slow release of ascorbate from erythrocytes can preserve both the plasma concentration of the vitamin, and prevent oxidation of alpha-tocopherol in low-density lipoprotein. In addition, intracellular ascorbate can spare and possibly recycle alpha-tocopherol in the erythrocyte membrane. In turn, alpha-tocopherol protects the cell membrane from lipid peroxidation. The ability of erythrocytes to recycle ascorbate, coupled with the ability of ascorbate to protect alpha-tocopherol in the cell membrane and in lipoproteins, provides a potentially important mechanism for preventing lipid peroxidative damage in areas of inflammation in the vascular bed, such as those involved with atherosclerosis. PMID- 9405335 TI - The p130 pocket protein: keeping order at cell cycle exit/re-entrance transitions. AB - Pocket proteins, including the retinoblastoma susceptibility gene product (pRB) and the related proteins p107 and p130, function at cell cycle regulatory steps that link cyclin/CDK-integrated positive and negative growth signals with E2F transcription factor activity on genes required for cell cycle progression. Protein complex formation between pocket proteins and members of the E2F family of transcription factors determines whether E2F complexes act as transcriptional activators or repressors. Experimental work over the last few years indicates that individual pocket proteins interact with specific E2F members to regulate the transcription of certain genes under diverse cell growth conditions. Among these protein associations, p130-containing E2F complexes seem to be of particular importance in controlling gene transcription in quiescent and differentiating cells by repressing the transcription of a set of E2F-responsive genes. Once the cells are progressing through the G1 phase of the cell cycle, pocket protein-mediated regulation of E2F activity is assumed by pRB and p107. p130-mediated transcriptional regulation thus seems to prevent a gene expression program characteristic of dividing cells at the cell cycle exit and re-entrance transitions and in quiescent cells. PMID- 9405336 TI - Signaling through scaffold, anchoring, and adaptor proteins. AB - The process by which extracellular signals are relayed from the plasma membrane to specific intracellular sites is an essential facet of cellular regulation. Many signaling pathways do so by altering the phosphorylation state of tyrosine, serine, or threonine residues of target proteins. Recently, it has become apparent that regulatory mechanisms exist to influence where and when protein kinases and phosphatases are activated in the cell. The role of scaffold, anchoring, and adaptor proteins that contribute to the specificity of signal transduction events by recruiting active enzymes into signaling networks or by placing enzymes close to their substrates is discussed. PMID- 9405337 TI - Large-cage zeolite structures with multidimensional 12-ring channels AB - Zeolite type structures with large cages interconnected by multidimensional 12 ring (rings of 12 tetrahedrally coordinated atoms) channels have been synthesized; more than a dozen large-pore materials were created in three different topologies with aluminum (or gallium), cobalt (or manganese, magnesium, or zinc), and phosphorus at the tetrahedral coordination sites. Tetragonal UCSB-8 has an unusually large cage built from 64 tetrahedral atoms and connected by an orthogonal channel system with 12-ring apertures in two dimensions and 8-ring apertures in the third. Rhombohedral UCSB-10 and hexagonal UCSB-6 are structurally related to faujasite and its hexagonal polymorph, respectively, and have large cages connected by 12-ring channels in all three dimensions. PMID- 9405338 TI - Immune versus natural selection: antibody aldolases with enzymic rates but broader scope. AB - Structural and mechanistic studies show that when the selection criteria of the immune system are changed, catalytic antibodies that have the efficiency of natural enzymes evolve, but the catalytic antibodies are much more accepting of a wide range of substrates. The catalytic antibodies were prepared by reactive immunization, a process whereby the selection criteria of the immune system are changed from simple binding to chemical reactivity. This process yielded aldolase catalytic antibodies that approximated the rate acceleration of the natural enzyme used in glycolysis. Unlike the natural enzyme, however, the antibody aldolases catalyzed a variety of aldol reactions and decarboxylations. The crystal structure of one of these antibodies identified the reactive lysine residue that was selected in the immunization process. This lysine is deeply buried in a hydrophobic pocket at the base of the binding site, thereby accounting for its perturbed pKa. PMID- 9405339 TI - Transcription regulation by initiating NTP concentration: rRNA synthesis in bacteria. AB - The sequence of a promoter determines not only the efficiency with which it forms a complex with RNA polymerase, but also the concentration of nucleoside triphosphate (NTP) required for initiating transcription. Escherichia coli ribosomal RNA (rrn P1) promoters require high initiating NTP concentrations for efficient transcription because they form unusually short-lived complexes with RNA polymerase; high initiating NTP concentrations [adenosine or guanosine triphosphate (ATP or GTP), depending on the rrn P1 promoter] are needed to bind to and stabilize the open complex. ATP and GTP concentrations, and therefore rrn P1 promoter activity, increase with growth rate. Because ribosomal RNA transcription determines the rate of ribosome synthesis, the control of ribosomal RNA transcription by NTP concentration provides a molecular explanation for the growth rate-dependent control and homeostatic regulation of ribosome synthesis. PMID- 9405340 TI - Multivariable dependence of Fe-Mg partitioning in the lower mantle AB - High-pressure diamond-cell experiments indicate that the iron-magnesium partitioning between (Fe,Mg)SiO3-perovskite and magnesiowustite in Earth's lower mantle depends on the pressure, temperature, bulk iron/magnesium ratio, and ferric iron content. The perovskite stability field expands with increasing pressure and temperature. The ferric iron component preferentially dissolves in perovskite and raises the apparent total iron content but had little effect on the partitioning of the ferrous iron. The ferrous iron depletes in perovskite at the top of the lower mantle and gradually increases at greater depth. These changes in iron-magnesium composition should affect geochemical and geophysical properties of the deep interior. PMID- 9405341 TI - Impact of molecular order in langmuir-blodgett films on catalysis AB - Catalytically active Langmuir-Blodgett films of a rhodium complex were prepared and characterized to determine the possible effect of the molecular order of metal complexes on catalytic activity. The hydrogenation of carbon-oxygen double bonds was used as a model reaction. The complex in solution exhibited low catalytic activity, whereas it was highly active in the film. The catalytic activity was found to be highly dependent on the orientation of the complex within the film. The reactions were also highly selective with regard to the substrate. These observations and the observed rate dependence on temperature strongly implicate the molecular order of a metal complex as an important dimension in catalysis. PMID- 9405342 TI - A tunable diode based on an inorganic Semiconductor&cjs3539;Conjugated polymer interface AB - Although in principle semiconductor-metal (Schottky) diodes should be tunable by changing the work function of the metal, such flexibility cannot be achieved in a single device and in practice is often limited by interfacial states that cause Fermi-level pinning. A tunable diode is reported based on a hybrid inorganic organic, n-indium phosphide&cjs3539;poly- (pyrrole)&cjs3539;nonaqueous electrolyte architecture. By electrochemically manipulating the work function of the conjugated polymer poly(pyrrole), the turn-on voltage (more precisely, the forward bias potential required to pass a particular current) of the diode can be continuously and actively tuned by more than 0.6 volt. The work highlights a distinguishing feature of conjugated polymers relative to more traditional semiconductor materials, namely, the ability of dopant ions to permeate conjugated polymers, thereby enabling electrochemical manipulation. PMID- 9405343 TI - Estimating the mass of asteroid 253 mathilde from tracking data during the NEAR flyby AB - The terminal navigation of the Near Earth Asteroid Rendezvous (NEAR) spacecraft during its close flyby of asteroid 253 Mathilde involved coordinated efforts to determine the heliocentric orbits of the spacecraft and Mathilde and then to determine the relative trajectory of the spacecraft with respect to Mathilde. The gravitational perturbation of Mathilde on the passing spacecraft was apparent in the spacecraft tracking data. As a result of the accurate targeting achieved, these data could be used to determine Mathilde's mass as 1.033 (+/- 0.044) x 10(20) grams. Coupled with a volume estimate provided by the NEAR imaging team, this mass suggests a low bulk density for Mathilde of 1.3 grams per cubic centimeter. PMID- 9405344 TI - NEAR's flyby of 253 mathilde: images of a C asteroid AB - On 27 June 1997, the Near Earth Asteroid Rendezvous (NEAR) spacecraft flew within 1212 kilometers of asteroid 253 Mathilde. Mathilde is an irregular, heavily cratered body measuring 66 kilometers by 48 kilometers by 46 kilometers. The asteroid's surface is dark (estimated albedo between 0.035 and 0.050) and similar in color to some CM carbonaceous chondrites. No albedo or color variations were detected. The volume derived from the images and the mass from Doppler tracking of the spacecraft yield a mean density of 1.3 +/- 0.2 grams per cubic centimeter, about half that of CM chondrites, indicating a porous interior structure. PMID- 9405345 TI - Quantum-confined stark effect in single CdSe nanocrystallite quantum dots AB - The quantum-confined Stark effect in single cadmium selenide (CdSe) nanocrystallite quantum dots was studied. The electric field dependence of the single-dot spectrum is characterized by a highly polarizable excited state ( approximately 10(5) cubic angstroms, compared to typical molecular values of order 10 to 100 cubic angstroms), in the presence of randomly oriented local electric fields that change over time. These local fields result in spontaneous spectral diffusion and contribute to ensemble inhomogeneous broadening. Stark shifts of the lowest excited state more than two orders of magnitude larger than the linewidth were observed, suggesting the potential use of these dots in electro-optic modulation devices. PMID- 9405346 TI - Natural variation in a Drosophila clock gene and temperature compensation. AB - The threonine-glycine (Thr-Gly) encoding repeat within the clock gene period of Drosophila melanogaster is polymorphic in length. The two major variants (Thr Gly)17 and (Thr-Gly)20 are distributed as a highly significant latitudinal cline in Europe and North Africa. Thr-Gly length variation from both wild-caught and transgenic individuals is related to the flies' ability to maintain a circadian period at different temperatures. This phenomenon provides a selective explanation for the geographical distribution of Thr-Gly lengths and gives a rare glimpse of the interplay between molecular polymorphism, behavior, population biology, and natural selection. PMID- 9405347 TI - Arabidopsis NPH1: a protein kinase with a putative redox-sensing domain. AB - The NPH1 (nonphototropic hypocotyl 1) gene encodes an essential component acting very early in the signal-transduction chain for phototropism. Arabidopsis NPH1 contains a serine-threonine kinase domain and LOV1 and LOV2 repeats that share similarity (36 to 56 percent) with Halobacterium salinarium Bat, Azotobacter vinelandii NIFL, Neurospora crassa White Collar-1, Escherichia coli Aer, and the Eag family of potassium-channel proteins from Drosophila and mammals. Sequence similarity with a known (NIFL) and a suspected (Aer) flavoprotein suggests that NPH1 LOV1 and LOV2 may be flavin-binding domains that regulate kinase activity in response to blue light-induced redox changes. PMID- 9405348 TI - Alignment of conduits for the nascent polypeptide chain in the ribosome-Sec61 complex. AB - An oligomer of the Sec61 trimeric complex is thought to form the protein conducting channel for protein transport across the endoplasmic reticulum. A purified yeast Sec61 complex bound to monomeric yeast ribosomes as an oligomer in a saturable fashion. Cryo-electron microscopy of the ribosome-Sec61 complex and a three-dimensional reconstruction showed that the Sec61 oligomer is attached to the large ribosomal subunit by a single connection. Moreover, a funnel-shaped pore in the Sec61 oligomer aligned with the exit of a tunnel traversing the large ribosomal subunit, strongly suggesting that both structures function together in the translocation of proteins across the endoplasmic reticulum membrane. PMID- 9405349 TI - Abscisic acid signaling through cyclic ADP-ribose in plants. AB - Abscisic acid (ABA) is the primary hormone that mediates plant responses to stresses such as cold, drought, and salinity. Single-cell microinjection experiments in tomato were used to identify possible intermediates involved in ABA signal transduction. Cyclic ADP-ribose (cADPR) was identified as a signaling molecule in the ABA response and was shown to exert its effects by way of calcium. Bioassay experiments showed that the amounts of cADPR in Arabidopsis thaliana plants increased in response to ABA treatment and before ABA-induced gene expression. PMID- 9405350 TI - Human factor IX transgenic sheep produced by transfer of nuclei from transfected fetal fibroblasts. AB - Ovine primary fetal fibroblasts were cotransfected with a neomycin resistance marker gene (neo) and a human coagulation factor IX genomic construct designed for expression of the encoded protein in sheep milk. Two cloned transfectants and a population of neomycin (G418)-resistant cells were used as donors for nuclear transfer to enucleated oocytes. Six transgenic lambs were liveborn: Three produced from cloned cells contained factor IX and neo transgenes, whereas three produced from the uncloned population contained the marker gene only. Somatic cells can therefore be subjected to genetic manipulation in vitro and produce viable animals by nuclear transfer. Production of transgenic sheep by nuclear transfer requires fewer than half the animals needed for pronuclear microinjection. PMID- 9405352 TI - Receptor-mediated protein kinase activation and the mechanism of transmembrane signaling in bacterial chemotaxis. AB - Chemotaxis responses of Escherichia coli and Salmonella are mediated by type I membrane receptors with N-terminal extracytoplasmic sensing domains connected by transmembrane helices to C-terminal signaling domains in the cytoplasm. Receptor signaling involves regulation of an associated protein kinase, CheA. Here we show that kinase activation by a soluble signaling domain construct involves the formation of a large complex, with approximately 14 receptor signaling domains per CheA dimer. Electron microscopic examination of these active complexes indicates a well defined bundle composed of numerous receptor filaments. Our findings suggest a mechanism for transmembrane signaling whereby stimulus-induced changes in lateral packing interactions within an array of receptor-sensing domains at the cell surface perturb an equilibrium between active and inactive receptor-kinase complexes within the cytoplasm. PMID- 9405351 TI - The 1.25 A crystal structure of sepiapterin reductase reveals its binding mode to pterins and brain neurotransmitters. AB - Sepiapterin reductase catalyses the last steps in the biosynthesis of tetrahydrobiopterin, the essential co-factor of aromatic amino acid hydroxylases and nitric oxide synthases. We have determined the crystal structure of mouse sepiapterin reductase by multiple isomorphous replacement at a resolution of 1.25 A in its ternary complex with oxaloacetate and NADP. The homodimeric structure reveals a single-domain alpha/beta-fold with a central four-helix bundle connecting two seven-stranded parallel beta-sheets, each sandwiched between two arrays of three helices. Ternary complexes with the substrate sepiapterin or the product tetrahydrobiopterin were studied. Each subunit contains a specific aspartate anchor (Asp258) for pterin-substrates, which positions the substrate side chain C1'-carbonyl group near Tyr171 OH and NADP C4'N. The catalytic mechanism of SR appears to consist of a NADPH-dependent proton transfer from Tyr171 to the substrate C1' and C2' carbonyl functions accompanied by stereospecific side chain isomerization. Complex structures with the inhibitor N acetyl serotonin show the indoleamine bound such that both reductase and isomerase activity for pterins is inhibited, but reaction with a variety of carbonyl compounds is possible. The complex structure with N-acetyl serotonin suggests the possibility for a highly specific feedback regulatory mechanism between the formation of indoleamines and pteridines in vivo. PMID- 9405353 TI - Efficient signal transduction by a chimeric yeast-mammalian G protein alpha subunit Gpa1-Gsalpha covalently fused to the yeast receptor Ste2. AB - Saccharomyces cerevisiae uses G protein-coupled receptors for signal transduction. We show that a fusion protein between the alpha-factor receptor (Ste2) and the Galpha subunit (Gpa1) transduces the signal efficiently in yeast cells devoid of the endogeneous STE2 and GPA1 genes. To evaluate the function of different domains of Galpha, a chimera between the N-terminal region of yeast Gpa1 and the C-terminal region of rat Gsalpha has been constructed. This chimeric Gpa1-Gsalpha is capable of restoring viability to haploid gpa1Delta cells, but signal transduction is prevented. This is consistent with evidence showing that the C-terminus of the homologous Galpha is required for receptor-G protein recognition. Surprisingly, a fusion protein between Ste2 and Gpa1-Gsalpha is able to transduce the signal efficiently. It appears, therefore, that the C-terminus of Galpha is mainly responsible for bringing the G protein into the close proximity of the receptor's intracellular domains, thus ensuring efficient coupling, rather than having a particular role in transmitting the signal. To confirm this conclusion, we show that two proteins interacting with each other (such as Snf1 and Snf4, or Ras and Raf), each of them fused either to the receptor or to the chimeric Galpha, allow efficient signal transduction. PMID- 9405354 TI - Induction of the cholesterol metabolic pathway regulates the farnesylation of RAS in embryonic chick heart cells: a new role for ras in regulating the expression of muscarinic receptors and G proteins. AB - We propose a novel mechanism for the regulation of the processing of Ras and demonstrate a new function for Ras in regulating the expression of cardiac autonomic receptors and their associated G proteins. We have demonstrated previously that induction of endogenous cholesterol synthesis in cultured cardiac myocytes resulted in a coordinated increase in expression of muscarinic receptors, the G protein alpha-subunit, G-alphai2, and the inward rectifying K+ channel, GIRK1. These changes in gene expression were associated with a marked increase in the response of heart cells to parasympathetic stimulation. In this study, we demonstrate that the induction of the cholesterol metabolic pathway regulates Ras processing and that Ras regulates expression of G-alphai2. We show that in primary cultured myocytes most of the RAS is localized to the cytoplasm in an unfarnesylated form. Induction of the cholesterol metabolic pathway results in increased farnesylation and membrane association of RAS. Studies of Ras mutants expressed in cultured heart cells demonstrate that activation of Ras by induction of the cholesterol metabolic pathway results in increased expression of G-alphai2 mRNA. Hence farnesylation of Ras is a regulatable process that plays a novel role in the control of second messenger pathways. PMID- 9405355 TI - The role of the linker between the SH2 domain and catalytic domain in the regulation and function of Src. AB - The crystal structures of the regulated Src and Hck tyrosine kinases show intramolecular interactions between the phosphorylated tail and the SH2 domain as well as between the SH3 domain, the SH2-catalytic domain linker (SH2-CD linker) and the catalytic domain. The relative contribution of these interactions to regulation of activity is poorly understood. Mutational analysis of Src and Lck revealed that interaction of the SH2-CD linker with the SH3 domain is crucial for regulation. Moreover, three sites of interaction of the linker with the catalytic domain, one at the beginning (Trp260) and two at the back of the small lobe, opposite the catalytic cleft (beta2/beta3 loop; alphaC/beta4 loop), impinge on Src activity. Other activating mutations map to the front of the catalytic domain in the loop preceding the alphaC-helix (beta3/alphaC loop). SH2-CD linker mutants are deregulated in mammalian cells but transform fibroblasts weakly, suggesting that the linker may bind cellular components. Interpretation of our results on the basis of the crystal structure of Src favours a model in which the correctly positioned SH2-CD linker exerts an inhibitory function on catalysis of Src family members by facilitating displacement of the alphaC-helix. This study may provide a template for the generation of deregulated versions of other protein kinases. PMID- 9405356 TI - Characterization of the ligand-binding site of the transferrin receptor in Trypanosoma brucei demonstrates a structural relationship with the N-terminal domain of the variant surface glycoprotein. AB - The Trypanosoma brucei transferrin (Tf) receptor is a heterodimer encoded by ESAG7 and ESAG6, two genes contained in the different polycistronic transcription units of the variant surface glycoprotein (VSG) gene. The sequence of ESAG7/6 differs slightly between different units, so that receptors with different affinities for Tf are expressed alternatively following transcriptional switching of VSG expression sites during antigenic variation of the parasite. Based on the sequence homology between pESAG7/6 and the N-terminal domain of VSGs, it can be predicted that the four blocks containing the major sequence differences between pESAG7 and pESAG6 form surface-exposed loops and generate the ligand-binding site. The exchange of a few amino acids in this region between pESAG6s encoded by different VSG units greatly increased the affinity for bovine Tf. Similar changes in other regions were ineffective, while mutations predicted to alter the VSG like structure abolished the binding. Chimeric proteins containing the N-terminal dimerization domain of VSG and the C-terminal half of either pESAG7 or pESAG6, which contains the ligand-binding domain, can form heterodimers that bind Tf. Taken together, these data provided evidence that the T.brucei Tf receptor is structurally related to the N-terminal domain of the VSG and that the ligand binding site corresponds to the exposed surface loops of the protein. PMID- 9405357 TI - A urokinase-sensitive region of the human urokinase receptor is responsible for its chemotactic activity. AB - The role of urokinase-type plasminogen activator (uPA) and its receptor (uPAR/CD87) in cell migration and invasion is well substantiated. Recently, uPA has been shown to be essential in cell migration, since uPA-/- mice are greatly impaired in inflammatory cell recruitment. We have shown previously that the uPA induced chemotaxis requires interaction with and modification of uPAR/CD87, which is the true chemoattracting molecule acting through an unidentified cell surface component which mediates this cell surface chemokine activity. By expressing and testing several uPAR/CD87 variants, we have located and functionally characterized a potent uPAR/CD87 epitope that mimics the effects of the uPA-uPAR interaction. The chemotactic activity lies in the region linking domains 1 and 2, the only protease-sensitive region of uPAR/CD87, efficiently cleaved by uPA at physiological concentrations. Synthetic peptides carrying this epitope promote chemotaxis and activate p56/p59(hck) tyrosine kinase. Both chemotaxis and kinase activation are pertussis toxin sensitive, involving a Gi/o protein in the pathway. PMID- 9405358 TI - Perforin is activated by a proteolytic cleavage during biosynthesis which reveals a phospholipid-binding C2 domain. AB - Perforin is a secreted protein synthesized by activated cytotoxic T lymphocytes (CTL) and natural killer (NK) cells. It is a key component of the lytic machinery of these cells, being able to insert into the plasma membrane of targeted cells, forming a pore which leads to their destruction. Here we analyse the synthesis, processing and intracellular transport of perforin in the NK cell line YT. Perforin is synthesized as a 70 kDa inactive precursor which is cleaved at the C terminus to yield a 60 kDa active form. This proteolytic cleavage occurs in an acidic compartment and can be inhibited by incubation of the cells in ammonium chloride, concanamycin A, leupeptin and E-64. The increased lytic activity of the cleaved form can be demonstrated by killing assays in which cleavage of the pro piece is inhibited. Epitope mapping reveals that cleavage of the pro-piece occurs at the boundary of a C2 domain, which we show is able to bind phospholipid membranes in a calcium-dependent manner. We propose that removal of the pro piece, which contains a bulky glycan, allows the C2 domain to interact with phospholipid membranes and initiate perforin pore formation. PMID- 9405359 TI - The catalytic cycle of the escherichia coli SecA ATPase comprises two distinct preprotein translocation events. AB - SecA is the ATP-dependent force generator in the Escherichia coli precursor protein translocation cascade, and is bound at the membrane surface to the integral membrane domain of the preprotein translocase. Preproteins are thought to be translocated in a stepwise manner by nucleotide-dependent cycles of SecA membrane insertion and de-insertion, or as large polypeptide segments by the protonmotive force (Deltap) in the absence of SecA. To determine the step size of a complete ATP- and SecA-dependent catalytic cycle, translocation intermediates of the preprotein proOmpA were generated at limiting SecA translocation ATPase activity. Distinct intermediates were formed, spaced by intervals of approximately 5 kDa. Inhibition of the SecA ATPase by azide trapped SecA in a membrane-inserted state and shifted the step size to 2-2.5 kDa. The latter corresponds to the translocation elicited by binding of non-hydrolysable ATP analogues to SecA, or by the re-binding of partially translocated polypeptide chains by SecA. Therefore, a complete catalytic cycle of the preprotein translocase permits the stepwise translocation of 5 kDa polypeptide segments by two consecutive events, i.e. approximately 2.5 kDa upon binding of the polypeptide by SecA, and another 2.5 kDa upon binding of ATP to SecA. PMID- 9405360 TI - The KDEL receptor, ERD2, regulates intracellular traffic by recruiting a GTPase activating protein for ARF1. AB - The small GTPase ADP-ribosylation factor 1 (ARF1) is a key regulator of intracellular membrane traffic. Regulators of ARF1, its GTPase-activating protein (GAP) and its guanine nucleotide exchange factor have been identified recently. However, it remains uncertain whether these regulators drive the GTPase cycle of ARF1 autonomously or whether their activities can be regulated by other proteins. Here, we demonstrate that the intracellular KDEL receptor, ERD2, self oligomerizes and interacts with ARF1 GAP, and thereby regulates the recruitment of cytosolic ARF1 GAP to membranes. Because ERD2 overexpression enhances the recruitment of GAP to membranes and results in a phenotype that reflects ARF1 inactivation, our findings suggest that ERD2 regulates ARF1 GAP, and thus regulates ARF1-mediated transport. PMID- 9405361 TI - Autocatalytic processing of the ATP-dependent PIM1 protease: crucial function of a pro-region for sorting to mitochondria. AB - The biogenesis of the ATP-dependent PIM1 protease of mitochondria was studied by mutational analysis. The ATPase and proteolytic activities of PIM1 were shown to be essential for mitochondrial function. A proteolytically inactive mutant form of PIM1 protease accumulated as a pro-form in mitochondria, revealing a two-step processing of PIM1: the matrix targeting signal is removed by the mitochondrial processing peptidase and then a pro-region of 61 amino acids is cleaved off in an autocatalytic reaction. This latter process depended on the ATP-dependent assembly of PIM1 protease subunits and can occur by an intermolecular and, most probably, also an intramolecular pathway. The respiratory competence of cells harboring mutant PIM1 protease lacking the pro-region was strongly impaired. Subcellular fractionation revealed a cytosolic localization of mutant PIM1 protease. This demonstrates the requirement for the propeptide for efficient sorting of PIM1 protease to mitochondria. PMID- 9405362 TI - Overexpression of Pex15p, a phosphorylated peroxisomal integral membrane protein required for peroxisome assembly in S.cerevisiae, causes proliferation of the endoplasmic reticulum membrane. AB - We have cloned PEX15 which is required for peroxisome biogenesis in Saccharomyces cerevisiae. pex15Delta cells are characterized by the cytosolic accumulation of peroxisomal matrix proteins containing a PTS1 or PTS2 import signal, whereas peroxisomal membrane proteins are present in peroxisomal remnants. PEX15 encodes a phosphorylated, integral peroxisomal membrane protein (Pex15p). Using multiple in vivo methods to determine the topology, Pex15p was found to be a tail-anchored type II (Ncyt-Clumen) peroxisomal membrane protein with a single transmembrane domain near its carboxy-terminus. Overexpression of Pex15p resulted in impaired peroxisome assembly, and caused profound proliferation of the endoplasmic reticulum (ER) membrane. The lumenal carboxy-terminal tail of Pex15p protrudes into the lumen of these ER membranes, as demonstrated by its O-glycosylation. Accumulation in the ER was also observed at an endogenous expression level when Pex15p was fused to the N-terminus of mature invertase. This resulted in core N glycosylation of the hybrid protein. The lumenal C-terminal tail of Pex15p is essential for targeting to the peroxisomal membrane. Furthermore, the peroxisomal membrane targeting signal of Pex15p overlaps with an ER targeting signal on this protein. These results indicate that Pex15p may be targeted to peroxisomes via the ER, or to both organelles. PMID- 9405363 TI - The chloroplastic protein import machinery contains a Rieske-type iron-sulfur cluster and a mononuclear iron-binding protein. AB - Transport of precursor proteins across the chloroplastic envelope membranes requires the interaction of protein translocons localized in both the outer and inner envelope membranes. Analysis by blue native gel electrophoresis revealed that the translocon of the inner envelope membranes consisted of at least six proteins with molecular weights of 36, 45, 52, 60, 100 and 110 kDa, respectively. Tic110 and ClpC, identified as components of the protein import apparatus of the inner envelope membrane, were prominent constituents of this complex. The amino acid sequence of the 52 kDa protein, deduced from the cDNA, contains a predicted Rieske-type iron-sulfur cluster and a mononuclear iron-binding site. Diethylpyrocarbonate, a Rieske-type protein-modifying reagent, inhibits the translocation of precursor protein across the inner envelope membrane, whereas binding of the precursor to the outer envelope membrane is still possible. In another independent experimental approach, the 52 kDa protein could be co purified with a trapped precursor protein in association with the chloroplast protein translocon subunits Toc86, Toc75, Toc34 and Tic110. Together, these results strongly suggest that the 52 kDa protein, named Tic55 due to its calculated molecular weight, is a member of the chloroplastic inner envelope protein translocon. PMID- 9405364 TI - Reconstitution of a chloroplast protein import channel. AB - The chloroplastic outer envelope protein OEP75 with a molecular weight of 75 kDa probably forms the central pore of the protein import machinery of the outer chloroplastic membrane. Patch-clamp analysis shows that heterologously expressed, purified and reconstituted OEP75 constitutes a voltage-gated ion channel with a unit conductance of Lambda = 145pS. Activation of the OEP75 channel in vitro is completely dependent on the magnitude and direction of the voltage gradient. Therefore, movements of protein charges of parts of OEP75 in the membrane electric field are required either for pore formation or its opening. In the presence of precursor protein from only one side of the bilayer, strong flickering and partial closing of the channel was observed, indicating a specific interaction of the precursor with OEP75. The comparatively low ionic conductance of OEP75 is compatible with a rather narrow aqueous pore (dporeapproximately equal to 8-9 A). Provided that protein and ion translocation occur through the same pore, this implies that the environment of the polypeptide during the transit is mainly hydrophilic and that protein translocation requires almost complete unfolding of the precursor. PMID- 9405365 TI - The novel SAR-binding domain of scaffold attachment factor A (SAF-A) is a target in apoptotic nuclear breakdown. AB - The scaffold attachment factor A (SAF-A) is an abundant component of the nuclear scaffold and of chromatin, and also occurs in heterogeneous nuclear ribonucleoprotein (hnRNP) complexes. Evidence from previous experiments had suggested that SAF-A most likely has at least two different functions, being involved both in nuclear architecture and RNA metabolism. We now show that the protein has a novel scaffold-associated region (SAR)-specific bipartite DNA binding domain which is independent from the previously identified RNA-binding domain, the RGG box. During apoptosis, but not during necrosis, SAF-A is cleaved in a caspase-dependent way. Cleavage occurs within the bipartite DNA-binding domain, resulting in a loss of DNA-binding activity and a concomitant detachment of SAF-A from nuclear structural sites. On the other hand, cleavage does not compromise the association of SAF-A with hnRNP complexes, indicating that the function of SAF-A in RNA metabolism is not affected in apoptosis. Our results suggest that detachment of SAF-A from SARs, caused by apoptotic proteolysis of its DNA-binding domain, is linked to the formation of oligonucleosomal-sized DNA fragments and could therefore contribute to nuclear breakdown in apoptotic cells. PMID- 9405366 TI - Reaper-induced apoptosis in a vertebrate system. AB - The reaper protein of Drosophila melanogaster has been shown to be a central regulator of apoptosis in that organism. However, it has not been shown to function in any vertebrate nor have the cellular components required for its action been defined. In this report we show that reaper can induce rapid apoptosis in vitro using an apoptotic reconstitution system derived from Xenopus eggs. Moreover, we show that a subcellular fraction enriched in mitochondria is required for this process and that reaper, acting in conjunction with cytosolic factors, can trigger mitochondrial cytochrome c release. Bcl-2 antagonizes these effects, but high levels of reaper can overcome the Bcl-2 block. These results demonstrate that reaper can function in a vertebrate context, suggesting that reaper-responsive factors are conserved elements of the apoptotic program. PMID- 9405368 TI - Defective dorsal closure and loss of epidermal decapentaplegic expression in Drosophila fos mutants. AB - Drosophila kayak mutant embryos exhibit defects in dorsal closure, a morphogenetic cell sheet movement during embryogenesis. Here we show that kayak encodes D-Fos, the Drosophila homologue of the mammalian proto-oncogene product, c-Fos. D-Fos is shown to act in a similar manner to Drosophila Jun: in the cells of the leading edge it is required for the expression of the TGFbeta-like Decapentaplegic (Dpp) protein, which is believed to control the cell shape changes that take place during dorsal closure. Defects observed in mutant embryos, and adults with reduced Fos expression, are reminiscent of phenotypes caused by 'loss of function' mutations in the Drosophila JNKK homologue, hemipterous. These results indicate that D-Fos is required downstream of the Drosophila JNK signal transduction pathway, consistent with a role in heterodimerization with D-Jun, to activate downstream targets such as dpp. PMID- 9405367 TI - Induction of TNF-sensitive cellular phenotype by c-Myc involves p53 and impaired NF-kappaB activation. AB - Normal fibroblasts are resistant to the cytotoxic action of tumor necrosis factor (TNF), but are rendered TNF-sensitive upon deregulation of c-Myc. To assess if oncoproteins induce the cytotoxic TNF activity by modulating TNF signaling, we investigated the TNF-elicited signaling responses in fibroblasts containing a conditionally active c-Myc protein. In association with cell death, c-Myc impaired TNF-induced activation of phospholipase A2, JNK protein kinase and cell survival-signaling-associated NF-kappaB transcription factor complex. The TNF induced death of mouse primary fibroblasts expressing deregulated c-Myc was inhibited by transient overexpression of the p65 subunit of NF-kappaB, which increased NF-kappaB activity in the cells. Unlike other TNF-induced signals, TNF induced accumulation of the wild-type p53 mRNA and protein was not inhibited by c Myc. TNF, with c-Myc, induced apoptosis in mouse primary fibroblasts but only weakly in p53-deficient primary fibroblasts. The C-terminal domain of p53, which is a transacting dominant inhibitor of wild-type p53, failed to inhibit apoptosis by c-Myc and TNF, suggesting that the cell death was not dependent on the transcription-activating function of p53. Taken together, the present findings show that the cytotoxic activity of TNF towards oncoprotein-expressing cells involves p53 and an impaired signaling for survival in such cells. PMID- 9405369 TI - Synergistic activation of a Drosophila enhancer by HOM/EXD and DPP signaling. AB - The homeotic proteins encoded by the genes of the Drosophila HOM and the vertebrate HOX complexes do not bind divergent DNA sequences with a high selectivity. In vitro, HOM (HOX) specificity can be increased by the formation of heterodimers with Extradenticle (EXD) or PBX homeodomain proteins. We have identified a single essential Labial (LAB)/EXD-binding site in a Decapentaplegic (DPP)-responsive enhancer of the homeotic gene lab which drives expression in the developing midgut. We show that LAB and EXD bind cooperatively to the site in vitro, and that the expression of the enhancer in vivo requires exd and lab function. In addition, point mutations in either the EXD or the LAB subsite compromise enhancer function, strongly suggesting that EXD and LAB bind to this site in vivo. Interestingly, we found that the activity of the enhancer is only stimulated by DPP signaling significantly upon binding of LAB and EXD. Thus, the enhancer appears to integrate positional information via the homeotic gene lab, and spatiotemporal information via DPP signaling; only when these inputs act in concert in an endodermal cell is the enhancer fully active. Our results illustrate how a tissue-specific response to DPP can be generated through synergistic effects on an enhancer carrying both DPP- and HOX-responsive sequences. PMID- 9405370 TI - XSmad2 directly activates the activin-inducible, dorsal mesoderm gene XFKH1 in Xenopus embryos. AB - Transforming growth factor (TGF)-beta family members play a central role in mesoderm induction during early embryogenesis in Xenopus. Although a number of target genes induced as an immediate-early response to activin-like members of the family have been described, little is known about the molecular mechanisms involved. Our systematic analysis of the activin induction of the target gene XFKH1 reveals two regions that mediate activin-responsive transcription: one, in the first intron, is targeted directly by the activin-signalling pathway; the other, in the 5' flanking sequences, responds to activin indirectly, possibly being required for maintenance of gene expression. We demonstrate that a 107 bp region of the XFKH1 first intron acts as an enhancer and confers activin inducibility onto a minimal uninducible promoter in the absence of new protein synthesis. It bears little sequence similarity to other activin responsive sequences. We further demonstrate that overexpression of a constitutively active derivative of Xenopus Smad2 (XSmad2), which has been implicated as a component of the activin signalling pathway, is sufficient for direct activation of transcription via this enhancer. Moreover, we show that XSmad2 acts indirectly on the proximal promoter element induced by activin via an indirect mechanism. These results establish the XFKH1 intron enhancer as a direct nuclear target of the activin signalling pathway in Xenopus embryos, and provide strong new evidence that XSmad2 is a transducer of activin signals. PMID- 9405371 TI - Position-dependent and promoter-specific regulation of gene expression in Trypanosoma brucei. AB - Trypanosoma brucei evades the mammalian immune response by a process of antigenic variation. This involves mutually exclusive and alternating expression of telomere-proximal variant surface glycoprotein genes (vsgs), which is controlled at the level of transcription. To examine transcription repression in T.brucei we inserted reporter genes, under the control of either rRNA or vsg expression site (ES) promoters, into various chromosomal loci. Position-dependent repression of both promoters was observed in the mammalian stage of the life cycle (bloodstream forms). Repression of promoters inserted into a silent ES was more pronounced closer to the telomere and was bi-directional. Transcription from both ES and rRNA promoters was also efficiently repressed at a non-telomeric vsg locus in bloodstream-form trypanosomes. In cultured tsetse fly midgut-stage (procyclic) trypanosomes, in which vsg is not normally expressed, all inserted rRNA promoters were derepressed but ES promoters remained silent. Our results suggest that vsg promoters and ectopic rRNA promoters in bloodstream-form T.brucei are restrained by position effects related to their proximity to vsgs or other features of the ES. Sequences present in rRNA promoters but absent from vsg ES promoters appear to be responsible for rRNA promoter-specific derepression in procyclic cells. PMID- 9405372 TI - Defective adipocyte differentiation in mice lacking the C/EBPbeta and/or C/EBPdelta gene. AB - To investigate the role of C/EBP family members during adipocyte differentiation in vivo, we have generated mice lacking the C/EBPbeta and/or C/EBPdelta by gene targeting. Approximately 85% of C/EBPbeta(-/-).delta(-/-) mice died at the early neonatal stage. By 20 h after birth, brown adipose tissue of the interscapular region in wild-type mice contained many lipid droplets, whereas C/EBPbeta(-/ ).delta(-/-) mice did not accumulate droplets. In addition, the epidydimal fat pad weight of surviving adult C/EBPbeta(-/-).delta(-/-) mice was significantly reduced compared with wild-type mice. However, these adipose tissues in C/EBPbeta(-/-).delta(-/-) mice exhibit normal expression of C/EBPalpha and PPARgamma, despite impaired adipogenesis. These results demonstrated that C/EBPbeta and C/EBPdelta have a synergistic role in terminal adipocyte differentiation in vivo. The induction of C/EBPalpha and PPARgamma does not always require C/EBPbeta and C/EBPdelta, but co-expression of C/EBPalpha and PPARgamma is not sufficient for complete adipocyte differentiation in the absence of C/EBPbeta and C/EBPdelta. PMID- 9405373 TI - Cisplatin- and UV-damaged DNA lure the basal transcription factor TFIID/TBP. AB - A connection between transcription and DNA repair was demonstrated previously through the characterization of TFIIH. Using filter binding as well as in vitro transcription challenge competition assays, we now show that the promoter recognition factor TATA box-binding protein (TBP)/TFIID binds selectively to and is sequestered by cisplatin- or UV-damaged DNA, either alone or in the context of a larger protein complex including TFIIH. Computer-assisted 3D structural analysis reveals a remarkable similarity between the structure of the TATA box as found in its TBP complex and that of either platinated or UV-damaged oligonucleotides. Thus, cisplatin-treated or UV-irradiated DNA could be used as a competing binding site which may lure TBP/TFIID away from its normal promoter sequence, partially explaining the phenomenon of DNA damage-induced inhibition of RNA synthesis. Consistent with an involvement of damaged DNA-specific binding of TBP in inhibiting transcription, we find that microinjection of additional TBP in living human fibroblasts alleviates the reduction in RNA synthesis after UV irradiation. Future anticancer drugs could be designed with the consideration of lesion recognition by TBP and their ability to reduce transcription. PMID- 9405374 TI - Multiple ATP-dependent steps in RNA polymerase II promoter melting and initiation. AB - Permanganate probing and abortive initiation assays were used to investigate the role of ATP in several successive stages of transcription initiation at the activated adeno E4 and mouse DHFR promoters. Removal of ATP at several points along the multi-step pathway blocked further progress towards its completion. Most strikingly, even if the DNA transcription start site is opened using ATP, the subsequent removal of ATP disallows formation of the first phosphodiester bond of the RNA. After ATP-dependent formation of a short RNA, a new transcription complex forms, which is more stable and has a longer open region. Both RNA and ATP appear to play roles in the formation of this complex. The need for ATP throughout this multi-step initiation pathway leads to new and unexpected possibilities for the use of energy and ATPases in transcription initiation. PMID- 9405376 TI - Ion-induced folding of the hammerhead ribozyme: a fluorescence resonance energy transfer study. AB - The ion-induced folding transitions of the hammerhead ribozyme have been analysed by fluorescence resonance energy transfer. The hammerhead ribozyme may be regarded as a special example of a three-way RNA junction, the global structure of which has been studied by comparing the distances (as energy transfer efficiencies) between the ends of pairs of labelled arms for the three possible end-to-end vectors as a function of magnesium ion concentration. The data support two sequential ion-dependent transitions, which can be interpreted in the light of the crystal structures of the hammerhead ribozyme. The first transition corresponds to the formation of a coaxial stacking between helices II and III; the data can be fully explained by a model in which the transition is induced by a single magnesium ion which binds with an apparent association constant of 8000 10 000 M-1. The second structural transition corresponds to the formation of the catalytic domain of the ribozyme, induced by a single magnesium ion with an apparent association constant of approximately 1100 M-1. The hammerhead ribozyme provides a well-defined example of ion-dependent folding in RNA. PMID- 9405375 TI - Three transitions in the RNA polymerase II transcription complex during initiation. AB - We have analyzed transcription initiation by RNA polymerase II (pol II) in a highly efficient in vitro transcription system composed of essentially homogeneous protein preparations. The pol II complex was stalled on adenovirus major late promoter templates at defined positions, and the open region and RNA products of these complexes were examined. The first transition is formation of the open complex, which can be reversed by addition of ATPgammaS. The open region is no longer sensitive to ATPgammaS after formation of a four-nucleotide RNA, which constitutes the second transition. This indicates that the ATP-dependent DNA helicase activity of TFIIH is required to maintain the open region only during formation of the first three phosphodiester bonds. The downstream part of the transcription bubble expands in a continuous motion, but the initially opened region (-9/-2 on the non-template strand) recloses abruptly when transcription reaches register 11. This third transition is accompanied by a switch from abortive to productive RNA synthesis, which implies promoter clearance. Our findings provide a framework to analyze regulation of these specific transitions during transcription initiation by pol II. PMID- 9405377 TI - Solution structure of a GAAA tetraloop receptor RNA. AB - The GAAA tetraloop receptor is an 11-nucleotide RNA sequence that participates in the tertiary folding of a variety of large catalytic RNAs by providing a specific binding site for GAAA tetraloops. Here we report the solution structure of the isolated tetraloop receptor as solved by multidimensional, heteronuclear magnetic resonance spectroscopy. The internal loop of the tetraloop receptor has three adenosines stacked in a cross-strand or zipper-like fashion. This arrangement produces a high degree of base stacking within the asymmetric internal loop without extrahelical bases or kinking the helix. Additional interactions within the internal loop include a U. U mismatch pair and a G.U wobble pair. A comparison with the crystal structure of the receptor RNA bound to its tetraloop shows that a conformational change has to occur upon tetraloop binding, which is in good agreement with previous biochemical data. A model for an alternative binding site within the receptor is proposed based on the NMR structure, phylogenetic data and previous crystallographic structures of tetraloop interactions. PMID- 9405378 TI - The constitutive transport element (CTE) of Mason-Pfizer monkey virus (MPMV) accesses a cellular mRNA export pathway. AB - The constitutive transport elements (CTEs) of type D retroviruses are cis-acting elements that promote nuclear export of incompletely spliced mRNAs. Unlike the Rev response element (RRE) of human immunodeficiency virus type 1 (HIV-1), CTEs depend entirely on factors encoded by the host cell genome. We show that an RNA comprised almost entirely of the CTE of Mason-Pfizer monkey virus (CTE RNA) is exported efficiently from Xenopus oocyte nuclei. The CTE RNA and an RNA containing the RRE of HIV-1 (plus Rev) have little effect on export of one another, demonstrating differences in host cell requirements of these two viral mRNA export pathways. Surprisingly, even very low amounts of CTE RNA block export of normal mRNAs, apparently through the sequestration of cellular mRNA export factors. Export of a CTE-containing lariat occurs when wild-type CTE, but not a mutant form, is inserted into the pre-mRNA. The CTE has two symmetric structures, either of which supports export and the titration of mRNA export factors, but both of which are required for maximal inhibition of mRNA export. Two host proteins bind specifically to the CTE but not to non-functional variants, making these proteins candidates for the sequestered mRNA export factors. PMID- 9405379 TI - A large nucleoprotein assembly at the ends of the viral DNA mediates retroviral DNA integration. AB - We have probed the nucleoprotein organization of Moloney murine leukemia virus (MLV) pre-integration complexes using a novel footprinting technique that utilizes a simplified in vitro phage Mu transposition system. We find that several hundred base pairs at each end of the viral DNA are organized in a large nucleoprotein complex, which we call the intasome. This structure is not formed when pre-integration complexes are made by infecting cells with integrase-minus virus, demonstrating a requirement for integrase. In contrast, footprinting of internal regions of the viral DNA did not reveal significant differences between pre-integration complexes with and without integrase. Treatment with high salt disrupts the intasome in parallel with loss of intermolecular integration activity. We show that a cellular factor is required for reconstitution of the intasome. Finally, we demonstrate that DNA-protein interactions involving extensive regions at the ends of the viral DNA are functionally important for retroviral DNA integration activity. Current in vitro integration systems utilizing purified integrase lack the full fidelity of the in vivo reaction. Our results indicate that both host factors and long viral DNA substrates may be required to reconstitute an in vitro system with all the hallmarks of DNA integration in vivo. PMID- 9405380 TI - Invertebrate retroviruses: ZAM a new candidate in D.melanogaster. AB - ZAM, a new retroelement of Drosophila melanogaster, was identified as a mutational insertion at the white locus. It displays all the structural features of a vertebrate retrovirus. Its three open reading frames encode predicted products resembling the products of the gag, pol and env genes of retroviruses. Its transcription gives rise to an 8.6 kb full-length RNA and a 1.7 kb spliced message for the env gene. The latter encodes an envelope protein that is typical of elements having an extracellular phase of the life cycle. The identification of a ZAM envelope retrogene provides evidence that ZAM is mobilized through a reverse trancriptional process in the germ line of flies. We report that ZAM is distributed differently among D.melanogaster strains. Two stocks out of >15 tested display a ZAM high copy number, with numerous copies distributed on chromosomal arms. This high copy number is associated with a high transcriptional rate of ZAM. The existence of these two categories of strains offers a new genetic system in which the properties of a potential invertebrate retrovirus can be tested. PMID- 9405381 TI - Solution structure of the Mu end DNA-binding ibeta subdomain of phage Mu transposase: modular DNA recognition by two tethered domains. AB - The phage Mu transposase (MuA) binds to the ends of the Mu genome during the assembly of higher order nucleoprotein complexes. We investigate the structure and function of the MuA end-binding domain (Ibetagamma). The three-dimensional solution structure of the Ibeta subdomain (residues 77-174) has been determined using multidimensional NMR spectroscopy. It comprises five alpha-helices, including a helix-turn-helix (HTH) DNA-binding motif formed by helices 3 and 4, and can be subdivided into two interacting structural elements. The structure has an elongated disc-like appearance from which protrudes the recognition helix of the HTH motif. The topology of helices 2-4 is very similar to that of helices 1-3 of the previously determined solution structure of the MuA Igamma subdomain and to that of the homeodomain family of HTH DNA-binding proteins. We show that each of the two subdomains binds to one half of the 22 bp recognition sequence, Ibeta to the more conserved Mu end distal half (beta subsite) and Igamma to the Mu end proximal half (gamma subsite) of the consensus Mu end-binding site. The complete Ibetagamma domain binds the recognition sequence with a 100- to 1000-fold higher affinity than the two subdomains independently, indicating a cooperative effect. Our results show that the Mu end DNA-binding domain of MuA has a modular organization, with each module acting on a specific part of the 22 bp binding site. Based on the present binding data and the structures of the Ibeta and Igamma subdomains, a model for the interaction of the complete Ibetagamma domain with DNA is proposed. PMID- 9405382 TI - The molecular genetics of rodent single gene obesities. PMID- 9405383 TI - RAD51 interacts with the evolutionarily conserved BRC motifs in the human breast cancer susceptibility gene brca2. AB - Recent work has shown that the murine BRCA2 tumor suppressor protein interacts with the murine RAD51 protein. This interaction suggests that BRCA2 participates in DNA repair. Residues 3196-3232 of the murine BRCA2 protein were shown to be involved in this interaction. Here, we report the detailed mapping of additional domains that are involved in interactions between the human homologs of these two proteins. Through yeast two-hybrid and biochemical assays, we demonstrate that the RAD51 protein interacts specifically with the eight evolutionarily conserved BRC motifs encoded in exon 11 of brca2 and with a similar motif found in a Caenorhabditis elegans hypothetical protein. Deletion analysis demonstrates that residues 98-339 of human RAD51 interact with the 59-residue minimal region that is conserved in all BRC motifs. These data suggest that the BRC repeats function to bind RAD51. PMID- 9405384 TI - Identification of residues in the drug-binding site of human P-glycoprotein using a thiol-reactive substrate. AB - We identified a thiol-reactive compound, dibromobimane (dBBn), that was a potent stimulator (8.2-fold) of the ATPase activity of Cys-less P-glycoprotein. We then used this compound together with cysteine-scanning mutagenesis to identify residues in transmembrane segment (TM) 6 and TM12 that are important for function. TM6 and TM12 lie close to each other in the tertiary structure and are postulated to be important for drug-protein interactions. The majority of P glycoprotein mutants containing a single cysteine residue retained substantial amounts of drug-stimulated ATPase activity and were not inhibited by dBBn. The ATPase activities of mutants L339C, A342C, L975C, V982C, and A985C, however, were markedly inhibited (>60%) by dBBn. The drug substrates verapamil, vinblastine, and colchicine protected these mutants against inhibition by dBBn, suggesting that these residues are important for interaction of substrates with P glycoprotein. We previously showed that residues Leu339, Ala342, Leu975, Val982, and Ala985 lie along the point of contact between helices TM6 and TM12, when both are aligned in a left-handed coiled coil (Loo, T. W., and Clarke, D. M. (1997) J. Biol. Chem. 272, 20986-20989). Taken together, these results suggest that the interface between TM6 and TM12 likely forms part of the potential drug-binding pocket in P-glycoprotein. PMID- 9405385 TI - Cloning and expressional characterization of a novel galanin receptor. Identification of different pharmacophores within galanin for the three galanin receptor subtypes. AB - Galanin, a 29-30 amino acid neuropeptide, is found in the central and peripheral nervous systems and displays several important physiological activities. The actions are believed to be mediated through distinct G protein-coupled receptors. To date, two galanin receptor subtypes have been cloned. In this report, we describe the cloning and expression of a cDNA encoding a novel galanin receptor (GalR3). The receptor has 370 amino acids and shares 36 and 54% homology with the rat GalR1 and GalR2 receptors. 125I-Porcine galanin binds the rat GalR3 receptor expressed in COS-7 cells with high affinity (Kd = 0.6 nM) and could be displaced by galanin and galanin fragments and galanin-chimeric peptides. The pharmacological profile of this novel receptor is distinct from those of GalR1 and GalR2, revealing different pharmacophores within galanin for the three galanin receptor subtypes. Northern blot analysis showed expression in heart, spleen, and testis. Unlike GalR1 and GalR2, no expression of GalR3 was detectable in the brain, suggesting that GalR3 may mediate some of the peripheral functions of galanin. PMID- 9405386 TI - A reevaluation of substrate specificity of the rat cation transporter rOCT1. AB - The substrate specificity of the previously cloned rat cation transporter rOCT1, which is expressed in kidney, liver, and small intestine, was reevaluated. rOCT1 is the first member of a new protein family comprising electrogenic and polyspecific cation transporters that transport hydrophilic cations like tetraethylammonium, choline, and monoamine neurotransmitters. Previous electrical measurements suggested that cations like quinine, quinidine, and cyanine 863, which have been classified as type 2 cations in the liver, are also transported by rOCT1, since they may induce inward currents in rOCT1 expressing Xenopus oocytes (Busch, A. E., Quester, S., Ulzheimer, J. C., Waldegger, S., Gorboulev, V., Arndt, P., Lang, F., and Koepsell, H. (1996) J. Biol. Chem. 271, 32599 32604). Tracer flux measurements with oocytes and with stably transfected human embryonic kidney cells showed that [3H]quinine and [3H]quinidine are not transported by rOCT1. The voltage dependence observed for the quinine- or quinidine-induced inward currents in rOCT1-expressing oocytes, and tracer efflux measurements indicate that the inward currents by type 2 cations are generated by the inhibition of electrogenic efflux of transported type 1 cations. Therefore, rOCT1 cannot contribute to transport of type 2 cations in the liver and the hepatic transporter for type 2 cations remains to be identified. PMID- 9405387 TI - Sulfated lewis X determinants as a major structural motif in glycans from LS174T HM7 human colon carcinoma mucin. AB - This article describes oligosaccharide structures of mucin isolated from nude mouse xenograft tumors produced by LS174T-HM7 cells, a subline of the human colon carcinoma LS174T with higher metastatic tendency and higher mucin production. A striking feature of the oligosaccharides of the LS174T-HM7 xenograft tumor mucin was a predominance of sulfated Lewis X determinants: HSO3-Galbeta1-4(Fucalpha1 3)GlcNAc. In addition to one previously known saccharide with one sulfated Lewis X determinant, the HM7 xenograft tumor mucin contained multiple novel structures containing one, two, or three sulfated Lewis X determinants. This determinant, known to act as a selectin ligand, has been found previously in minor saccharide components of human milk as well as mucins, but never before as a predominant structure in one mucin source. PMID- 9405388 TI - Cloning of a human purinergic P2Y receptor coupled to phospholipase C and adenylyl cyclase. AB - Clones encoding a new human P2Y receptor, provisionally called P2Y11, have been isolated from human placenta complementary DNA and genomic DNA libraries. The 1113-base pair open reading frame is interrupted by one intron. The P2Y11 receptor is characterized by considerably larger second and third extracellular loops than the subtypes described so far. The deduced amino acid sequence exhibits 33% amino acid identity with the P2Y1 receptor, its closest homolog. Northern blot analysis detected human P2Y11 receptor messenger RNA in spleen and HL-60 cells. The level of P2Y11 transcripts was strongly increased in these cells after granulocyte differentiation induced by retinoic acid or dimethyl sulfoxide. The new receptor was stably expressed in 1321N1 astrocytoma and CHO-K1 cells, where it couples to the stimulation of both the phosphoinositide and adenylyl cyclase pathways, a unique feature among the P2Y family. The rank order of agonists potency was: ATP > 2-methylthio-ATP >>> ADP, whereas UTP and UDP were inactive, indicating that it behaves as a selective purinoceptor. PMID- 9405389 TI - Cloning and expression of a cDNA encoding bovine lipoyltransferase. AB - Lipoyltransferase catalyzes the transfer of the lipoyl group from lipoyl-AMP to the specific lysine residue of the lipoate-dependent enzymes. We have isolated lipoyltransferase I (LipTI) and II (LipTII) from bovine liver (Fujiwara, K., Okamura-Ikeda, K., and Motokawa, Y. (1994) J. Biol. Chem. 269, 16605-16609). N terminal amino acid sequences of LipTI and LipTII were identical except that LipTI had an additional Asn residue on the N terminus. We cloned LipTII cDNA from a bovine liver cDNA library. The cDNA insert contained a 1119-base pair open reading frame encoding a precursor peptide of 373 amino acids including a mitochondrial targeting signal of 26 amino acids. The calculated molecular mass of the mature protein is 39,137 Da. The predicted amino acid sequence showed 35% identity with that of Escherichia coli lipoate-protein ligase A. Northern and Southern blot analyses showed a single band, and a single species of mRNA for lipoyltransferase was found by reverse transcription-polymerase chain reaction. Recombinant LipTII was expressed in E. coli and purified to apparent homogeneity. The Kmapp values of the recombinant enzyme for lipoyl-AMP and apoH-protein were comparable with those of native LipTII. An antibody raised against recombinant enzyme cross-reacted with LipTI and LipTII in a similar manner. The results suggest that LipTI and LipTII are derived from the same translated product but processed differently. PMID- 9405390 TI - A family of human beta4-galactosyltransferases. Cloning and expression of two novel UDP-galactose:beta-n-acetylglucosamine beta1, 4-galactosyltransferases, beta4Gal-T2 and beta4Gal-T3. AB - BLAST analysis of expressed sequence tags (ESTs) using the coding sequence of the human UDP-galactose:beta-N-acetylglucosamine beta1, 4-galactosyltransferase, designated beta4Gal-T1, revealed a large number of ESTs with identical as well as similar sequences. ESTs with sequences similar to that of beta4Gal-T1 could be grouped into at least two non-identical sequence sets. Analysis of the predicted amino acid sequence of the novel ESTs with beta4Gal-T1 revealed conservation of short sequence motifs as well as cysteine residues previously shown to be important for the function of beta4Gal-T1. The likelihood that the identified ESTs represented novel galactosyltransferase genes was tested by cloning and sequencing of the full coding region of two distinct genes, followed by expression. Expression of soluble secreted constructs in the baculovirus system showed that these genes represented genuine UDP-galactose:beta-N acetylglucosamine beta1, 4-galactosyltransferases, thus designated beta4Gal-T2 and beta4Gal-T3. Genomic cloning of the genes revealed that they have identical genomic organizations compared with beta4Gal-T1. The two novel genes were located on 1p32-33 and 1q23. The results demonstrate the existence of a family of homologous galactosyltransferases with related functions. The existence of multiple beta4-galactosyltransferases with the same or overlapping functions may be relevant for interpretation of biological functions previously assigned to beta4Gal-T1. PMID- 9405391 TI - Enzymatic characterization of human alpha1,3-fucosyltransferase Fuc-TVII synthesized in a B cell lymphoma cell line. AB - The human alpha1,3-fucosyltransferase, Fuc-TVII, a key enzyme in the biosynthesis of selectin ligands, was expressed as a soluble protein-A chimeric form in a human B cell lymphoma cell line, Namalwa KJM-1, and purified using IgG-Sepharose. The enzymatic properties of recombinant soluble Fuc-TVII were then examined. Its enzyme activity was highest at pH 7.5, and the presence of 25 mM Mn2+ was required for full activity. Fuc-TVII exhibits an acceptor specificity restricted to alpha2,3-sialylated type 2 oligosaccharides, and the apparent Km values for alpha2,3-sialyl lacto-N-neotetraose and GDP-fucose were 3.08 mM and 16.4 microM, respectively. The inhibitory effects of various nucleotides on the activity of Fuc-TVII reflected its donor specificity for the nucleotide portion of GDP. Fuc TVII was demonstrated to be useful for the synthesis of a sialyl Lewis x hexasaccharide from lacto-N-neotetraose in combination with an alpha2, 3 sialyltransferase, ST3Gal IV. Polyethylene glycols enhanced the thermal stability of Fuc-TVII, leading to increased formation of the reaction product. PMID- 9405392 TI - Dissecting the cooperative reassociation of the regulatory and catalytic subunits of cAMP-dependent protein kinase. Role of Trp-196 in the catalytic subunit. AB - The catalytic (C) subunit of cAMP-dependent protein kinase requires two distinct surfaces to form a stable complex with its physiological inhibitors, the regulatory (R) subunits and the heat-stable protein kinase inhibitors. In addition to a substrate-like segment that is common to both inhibitors, R requires a peripheral recognition site, PRS2. This surface is comprised of the essential phosphorylation site, Thr-197, His-87, Trp-196, and several surrounding basic residues. To probe the role of Trp-196 in the recognition of R, Trp-196 was replaced with Arg and Ala. Although both rC(W196A) and rC(W196R) were inhibited readily with cAMP-free R, they failed to form an inhibited holoenzyme complex with native R under conditions in which wild-type holoenzyme formed readily. Pairing rC(W196R) with mutant forms of R lacking domain B or having defects in cAMP binding sites A or B highlighted the importance of the conformation of R, and, in particular, the accessibility of site A. One of these mutants, rR(R333K), having a defect in cAMP binding site B formed a stable complex with rC(W196R) in the absence of cAMP. However, unlike wild-type holoenzyme, this complex was active. PMID- 9405393 TI - Cloning and tyrosine phosphorylation of a novel invertebrate immunocyte protein containing immunoreceptor tyrosine-based activation motifs. AB - Immunoreceptor tyrosine-based activation motif (ITAM) plays an important role in signal transduction through mammalian T-cell and B-cell antigen receptors and Fc receptors. The ITAM has been found only in vertebrate immunocytes. Ascidians are intriguing invertebrates from the viewpoint of the evolution of immune systems because they are considered to be ancestors of the vertebrates. We have previously shown that the monoclonal antibody A74 inhibits cellular defense reactions of the ascidian. In the present studies, we found that the A74 antigen protein has two ITAMs and several motifs that are proposed to function in signal transduction. The A74 protein is tyrosine-phosphorylated and associated with other proteins in the initial stages of cellular defense reactions. The ITAMs of the A74 protein are tyrosine-phosphorylated by a c-Src kinase in vitro. The A74 protein provides a key to the understanding of the origin of vertebrate immune systems. PMID- 9405394 TI - The serine/threonine phosphatase PP5 interacts with CDC16 and CDC27, two tetratricopeptide repeat-containing subunits of the anaphase-promoting complex. AB - The evolutionarily conserved multisubunit complex known as the cyclosome or anaphase-promoting complex is involved in catalyzing the ubiquitination of diverse substrates in M phase, allowing their destruction by the 26 S proteasome and the completion of mitosis. Three of the eight subunits of the anaphase promoting complex (CDC16, CDC23, and CDC27) have been shown to be phosphorylated in M phase, and their phosphorylation is required for the anaphase-promoting complex to be active as a ubiquitin ligase. Several subunits of the anaphase promoting complex contain tetratricopeptide repeats, a protein motif involved in protein/protein interactions. PP5 is a serine/threonine phosphatase that also contains four copies of the tetratricopeptide repeats motif. Here we show by a combination of two-hybrid analysis and in vitro binding that PP5 interacts with CDC16 and CDC27, two subunits of the anaphase-promoting complex. Only the NH2 terminal domain of PP5, containing all four tetratricopeptide repeats, is required for this physical interaction. Deletion analysis suggests that the site of binding to PP5 is localized to the COOH-terminal block of tetratricopeptide repeats in CDC16 and CDC27. In addition, indirect immunofluorescence showed that PP5 localizes to the mitotic spindle apparatus. The direct interaction of PP5 with CDC16 and CDC27, as well as its overlapping spindle localization in mitosis, suggests that PP5 may be involved in the regulation of the activity of the anaphase-promoting complex. PMID- 9405395 TI - Specific interaction of the PDZ domain protein PICK1 with the COOH terminus of protein kinase C-alpha. AB - PICK1 is a protein kinase C (PKC) alpha-binding protein initially identified using the yeast two-hybrid system. Here we report that PICK1 contains a PDZ domain that interacts specifically with a previously unidentified PDZ-binding domain (QSAV) at the extreme COOH terminus of PKCalpha and that mutation of a putative carboxylate-binding loop within the PICK1 PDZ domain abolishes this interaction. The PDZ-binding domain in PKCalpha is absent from other PKC isoforms that do not interact with PICK1. We also demonstrate that PICK1 can homooligomerize through sequences that are distinct from the carboxylate-binding loop, suggesting that self-association and PKCalpha binding are not mutually exclusive. A Caenorhabditis elegans PICK1-like protein is also able to bind to PKCalpha, suggesting a conservation of function through evolution. Association of PKCalpha with PICK1 provides a potential mechanism for the selective targeting of PKCalpha to unique subcellular sites. PMID- 9405396 TI - Cytosolic pyridoxine-beta-D-glucoside hydrolase from porcine jejunal mucosa. Purification, properties, and comparison with broad specificity beta-glucosidase. AB - During studies of the nutritional utilization of pyridoxine 5'-beta-D-glucoside, a major form of vitamin B6 in plants, we detected two cytosolic beta-glucosidases in jejunal mucosa. As expected, one was broad specificity beta-glucosidase that hydrolyzed aryl beta-D-glycosides but not pyridoxine beta-D-glucoside. We also found a previously unknown enzyme, designated pyridoxine-beta-D-glucoside hydrolase, that efficiently hydrolyzed pyridoxine beta-D-glucoside. These were separated and purified as follows: broad specificity beta-glucosidase 1460-fold and pyridoxine-beta-D-glucoside hydrolase 36,500-fold. Purified pyridoxine-beta-D glucoside hydrolase did not hydrolyze any of the aryl glycosides tested but did hydrolyze cellobiose and lactose. Pyridoxine-beta-D-glucoside hydrolase exhibited a pH optimum of 5.5 and apparent molecular mass of 130 kDa by SDS-polyacrylamide gel electrophoresis and 160 kDa by nondenaturing gel filtration, in contrast to 60 kDa for native and denatured broad specificity beta-glucosidase. Glucono-delta lactone was a strong inhibitor of both enzymes. Ionic and nonionic detergents were inhibitory for each enzyme. Conduritol B epoxide, a potent inhibitor of lysosomal acid beta-glucosidase, inhibited pyridoxine-beta-D-glucoside hydrolase but not broad specificity beta-glucosidase, but both were inhibited by the mechanism-based inhibitor 2-deoxy-2-fluoro-beta-D-glucosyl fluoride. Our findings indicate major differences between these two cytosolic beta-glucosidases. Studies addressing the role of vitamin B6 nutrition in regulating the activity and its consequences regarding pyridoxine glucoside bioavailability are in progress. PMID- 9405397 TI - Characterization, sequencing, and expression of the genes encoding a reactivating factor for glycerol-inactivated adenosylcobalamin-dependent diol dehydratase. AB - Diol dehydratase undergoes suicide inactivation by glycerol during catalysis involving irreversible cleavage of the Co-C bond of adenosylcobalamin. In permeabilized Klebsiella oxytoca and Klebsiella pneumoniae cells, the glycerol inactivated holoenzyme or the enzyme-cyanocobalamin complex is rapidly activated by the exchange of the inactivated coenzyme or cyanocobalamin for free adenosylcobalamin in the presence of ATP and Mg2+ (Honda, S., Toraya, T., and Fukui, S. (1980) J. Bacteriol. 143, 1458-1465; Ushio, K., Honda, S., Toraya, T., and Fukui, S. (1982) J. Nutr. Sci. Vitaminol. 28, 225-236). Permeabilized Escherichia coli cells co-expressing the diol dehydratase genes with two open reading frames in the 3'-flanking region were capable of reactivating glycerol inactivated diol dehydratase as well as activating the enzyme-cyanocobalamin complex in situ in the presence of free adenosylcobalamin, ATP, and Mg2+. These open reading frames, designated as ddrA and ddrB genes, were identified as the genes of a putative reactivating factor for inactivated diol dehydratase. The genes encoded polypeptides consisting of 610 and 125 amino acid residues with predicted molecular weights of 64,266 and 13,620, respectively. Co-expression of the open reading frame in the 5'-flanking region was stimulatory but not obligatory for conferring the reactivating activity upon E. coli. Thus, the product of this gene was considered not an essential component of the reactivating factor. PMID- 9405398 TI - Lipase activities of p37, the major envelope protein of vaccinia virus. AB - p37, the major protein of the extracellular enveloped form of vaccinia virus, is involved in the biogenesis of the viral double membrane and in egress of virus from the cell. p37 was expressed as a glutathione S-transferase fusion protein and was purified to homogeneity by silver staining using glutathione-agarose, Sephacryl S-200, and DEAE-cellulose chromatography. Incubation of p37 with phosphatidylcholine labeled in the fatty acyl side chains resulted in the production of multiple lipid products that were identified by thin layer chromatography and mass spectrometry as diacylglycerol, free fatty acid, monoacylglycerol, and lysophosphatidylcholine. Lipid-metabolizing activities colocalized with p37-containing fractions throughout the chromatographic steps. p37 also metabolized phosphatidylethanolamine efficiently, but it had less activity toward phosphatidylinositol and little or no activity toward phosphatidylserine. The purified enzyme also metabolized triacylglycerol to diacylglycerol but was inactive toward sn-1, 2-diacylglycerol. p37 was also expressed in insect cells as a poly-His fusion protein; cell lysates and partially purified proteins also generated products expected from phospholipase C and A activities. Thus, p37 is a broad specificity lipase with phospholipase C, phospholipase A, and triacylglycerol lipase activities. PMID- 9405399 TI - T state hemoglobin binds oxygen noncooperatively with allosteric effects of protons, inositol hexaphosphate, and chloride. AB - Hemoglobin is the paradigm of allosteric proteins. Over the years, cooperative oxygen binding has been explained by different models predicting that the T state of hemoglobin binds oxygen either noncooperatively or with some degree of cooperativity or with strong cooperativity. Therefore, a critical test that discriminates among models is to determine the oxygen binding by the T state of hemoglobin. Fixation of hemoglobin in the T state has been achieved either by crystallization from polyethylene glycol solutions or by encapsulation in wet porous silica gels. Hemoglobin crystals bind oxygen noncooperatively with reduced affinity compared with solution, with no Bohr effect and with no influence of other allosteric effectors. In this study, we have determined accurate oxygen binding curves to the T state of hemoglobin in silica gels with the same microspectrophotometric apparatus and multiwavelengths analysis used in crystal experiments. The T state of hemoglobin in silica gels binds oxygen noncooperatively with an affinity and a Bohr effect similar to those observed in solution for the binding of the first oxygen molecule. Other allosteric effectors such as inositol hexaphosphate, bezafibrate, and chloride significantly affect oxygen affinity. Therefore, T state hemoglobins that are characterized by strikingly different functional properties share the absence of cooperativity in the binding of oxygen. These findings are fully consistent with the Monod, Wyman, and Changeux model and with most features of Perutz's stereochemical model, but they are not consistent with models of both Koshland and Ackers. PMID- 9405400 TI - MEKK1 binds directly to the c-Jun N-terminal kinases/stress-activated protein kinases. AB - Mitogen-activated protein (MAP) kinases mediate responses to a wide array of cellular stimuli. These cascades consist of a MAP kinase or extracellular signal regulated kinase (ERK), activated by a MAP/ERK kinase (MEK), in turn activated by a MEK kinase (MEKK). MEKK1 has been shown to be a strong activator of the c-Jun N terminal kinase/stress-actived protein kinase (JNK/SAPK) pathway. We report here that JNK/SAPK binds directly to the N-terminal, noncatalytic domain of MEKK1 in vitro and in transfected cells. Immobilized MEKK1-derived peptides extract JNK/SAPK selectively from cell lysates. MEKK1 coimmunoprecipitates with multiple JNK/SAPK isoforms in transfected cells. Expression of the N terminus of MEKK1 lacking the kinase domain increases activation of endogenous JNK/SAPK by MEKK1. The data are consistent with a model in which MEKK1-JNK/SAPK binding facilitates the receipt of signals from upstream inputs and localizes JNK/SAPK to intracellular targets of the pathway. PMID- 9405401 TI - Alternative translation of the proto-oncogene c-myc by an internal ribosome entry site. AB - The human proto-oncogene c-myc encodes two proteins, c-Myc1 and c-Myc2, from two initiation codons, CUG and AUG, respectively. It is also transcribed from four alternative promoters (P0, P1, P2, and P3), giving rise to different RNA 5' leader sequences, the long sizes of which suggest that they must be inefficiently translated by the classical ribosome scanning mechanism. Here we have examined the influence of three c-myc mRNA 5'-leaders on the translation of chimeric myc CAT mRNAs. We observed that in the reticulocyte rabbit lysate, these 5'-leaders lead to cap-independent translation initiation. To determine whether this kind of initiation resulted from the presence of an internal ribosome entry site (IRES), COS-7 cells were transfected with bicistronic vectors containing the different c myc 5'-leaders in the intercistronic region. An IRES was identified, requiring elements located within the P2 leader, between nucleotides -363 and -94 upstream from the CUG start codon. This is the first demonstration of the existence of IRES-dependent translation for a proto-oncogene. This IRES could be a translation enhancer, allowing activation of c-myc expression under the control of trans acting factors and in response to specific cell stimuli. PMID- 9405402 TI - Does calponin interact with caldesmon? AB - The roles of calponin and caldesmon and their interaction in regulation of smooth muscle contraction are controversial. Recently, strong binding between these two proteins has been reported (Graceffa, P., Adam, L. P., and Morgan, K. G. (1996) J. Biol. Chem. 271, 30336-30339). Results in this paper fail to confirm their data and are consistent with the concept of independent functions for calponin and caldesmon. To examine the ability of duck gizzard caldesmon to interact with calponin, three caldesmon derivatives, each containing a different sulfhydryl specific reporter probe (6-acryloyl-2-dimethylaminonaphtalene, N-(1 pyrenyl)iodoacetamide, and N-iodoacetyl-N'-(5-sulfo-1-naphtylo)ethylenediamine) attached to a single cysteine located in the C-terminal domain, were synthesized. Addition of calponin to labeled caldesmon at both low and physiological salt concentrations did not induce any changes in fluorescence intensity or maximum shift. Under the same conditions, calmodulin and tropomyosin (known to bind to the C terminus of caldesmon) produced substantial changes in these spectral parameters. Gel filtration of an equimolar caldesmon-calponin mixture on a fast protein liquid chromatography Superose-12 column revealed two base-line-separated peaks, the first containing only caldesmon and the second only calponin, thus confirming the lack of any interaction between these two proteins. Also, the addition of calponin did not change the fluorescence parameters of labeled caldesmon in complexes with F-actin and F-actin-tropomyosin. PMID- 9405403 TI - Molecular determinants of selectivity in 5-hydroxytryptamine1B receptor-G protein interactions. AB - The recognition between G protein and cognate receptor plays a key role in specific cellular responses to environmental stimuli. Here we explore specificity in receptor-G protein coupling by taking advantage of the ability of the 5 hydroxytryptamine1B (5-HT1B) receptor to discriminate between G protein heterotrimers containing Galphai1 or Galphat. Gi1 can interact with the 5-HT1B receptor and stabilize a high affinity agonist binding state of this receptor, but Gt cannot. A series of Galphat/Galphai1 chimeric proteins have been generated in Escherichia coli, and their functional integrity has been reported previously (Skiba, N. P., Bae, H., and Hamm, H. E. (1996) J. Biol. Chem. 271, 413-424). We have tested the functional coupling abilities of the Galphat/Galphai1 chimeras to 5-HT1B receptors using high affinity agonist binding and receptor-stimulated guanosine 5'-3-O-(thio)triphosphate (GTPgammaS) binding. In the presence of betagamma subunits, amino acid residues 299-318 of Galphai1 increase agonist binding to the 5-HT1B receptor and receptor stimulation of GTPgammaS binding. Moreover, Galphai1 containing only Galphat amino acid sequences from this region does not show any coupling ability to 5-HT1B receptors. Our studies suggest that the alpha4 helix and alpha4-beta6 loop region of Galphas are an important region for specific recognition between receptors and Gi family members. PMID- 9405405 TI - Detection of noncovalent tRNA.aminoacyl-tRNA synthetase complexes by matrix assisted laser desorption/ionization mass spectrometry. AB - Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-MS) was used for the study of complexes formed by yeast seryl-tRNA synthetase (SerRS) and tyrosyl-tRNA synthetase (TyrRS) with tRNASer and tRNATyr. Cognate and noncognate complexes were easily distinguished due to a large mass difference between the two tRNAs. Both homodimeric synthetases gave MS spectra indicating intact desorption of dimers. The spectra of synthetase-cognate tRNA mixtures showed peaks of free components and peaks assigned to complexes. Noncognate complexes were also detected. In competition experiments, where both tRNA species were mixed with each enzyme only cognate alpha2.tRNA complexes were observed. Only cognate alpha2.tRNA2 complexes were detected with each enzyme. These results demonstrate that MALDI-MS can be used successfully for accurate mass and, thus, stoichiometry determination of specific high molecular weight noncovalent protein-nucleic acid complexes. PMID- 9405404 TI - Cloning and characterization of a novel promiscuous human beta-chemokine receptor D6. AB - Members of the chemokine family of chemotactic peptides interact with their target cells through heptahelical cell surface receptors. An understanding of the biochemistry and expression patterns of these receptors is therefore central to our overall understanding of the roles played by chemokines in both physiological and pathological processes. To date, eight receptors for the beta-chemokine subfamily have been described. We have recently cloned a novel murine beta chemokine receptor and report here the identification and characterization of a highly homologous human gene termed human D6 (hD6). This is a promiscuous beta chemokine receptor and appears to be able to bind the majority of members of the beta-chemokine family. It is, however, specific for this family and shows no detectable affinity for members of the alpha-chemokine or the C or CXXXC chemokines. Unlike the majority of other chemokine receptors, human D6 does not appear to be able to flux calcium following ligand binding, thus it is currently not clear if this novel receptor is indeed a signaling receptor. Human D6 is expressed in a range of tissues including hemopoietic cells although it appears not to be ubiquitously expressed in hemopoietic cells. Human D6, unlike some other beta-chemokine receptors, appears not to be able to function as an entry co factor for human immunodeficiency virus type 1 (HIV-)1 on CD4-expressing cells. PMID- 9405406 TI - Isolation of a tenascin-R binding protein from mouse brain membranes. A phosphacan-related chondroitin sulfate proteoglycan. AB - We have isolated a chondroitin sulfate proteoglycan from mouse brain by affinity chromatography with a fragment of the extracellular matrix glycoprotein tenascin R (TN-R) that comprises the amino-terminal cysteine-rich stretch and the 4.5 epidermal growth factor-like repeats. The isolated chondroitin sulfate proteoglycan has a molecular mass of 500-600 kDa and carries the HNK-1 carbohydrate epitope. Treatment with chondroitinase ABC reveals a major band of approximately 400 kDa and two minor bands at 200 and 150 kDa. Immunoblot analysis relates the molecule to phosphacan but not to the chondroitin sulfate proteoglycans neurocan and versican. Binding of the phosphacan-related molecule to the epidermal growth factor-like repeats of TN-R is Ca2+-dependent. Co localization of the molecule with TN-R in the retina and optic nerve by immunocytochemistry suggests a functional relationship between the two molecules in vivo. Inhibition of neurite outgrowth from hippocampal neurons by the phosphacan-related molecule in vitro is neutralized by TN-R when coated as a uniform substrate. Furthermore, the phosphacan-related molecule neutralizes growth cone repulsion induced by TN-R coated as a sharp substrate boundary with or without prior treatment with chondroitinase ABC. These observations indicate that TN-R can interact with a phosphacan-related molecule and thereby modulate its inhibitory influence on neuritogenesis. PMID- 9405407 TI - Activation of stress-activated protein kinase/c-Jun N-terminal kinase, but not NF kappaB, by the tumor necrosis factor (TNF) receptor 1 through a TNF receptor associated factor 2- and germinal center kinase related-dependent pathway. AB - A key step by which tumor necrosis factor (TNF) signals the activation of nuclear factor-kappaB (NF-kappaB) and the stress-activated protein kinase (SAPK, also called c-Jun N-terminal kinase or JNK) is the recruitment to the TNF receptor of TNF receptor-associated factor 2 (TRAF2). However, the subsequent steps in TRAF2 induced SAPK and NF-kappaB activation remain unresolved. Here we report the identification of a TNF-responsive serine/threonine protein kinase termed GCK related (GCKR) that likely signals via mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) kinase kinase 1 (MEKK1) to activate the SAPK pathway. TNF, TRAF2, and ultraviolet (UV) light, which in part uses the TNF receptor signaling pathway, all increased GCKR activity. A TRAF2 mutant, which inhibits both TRAF2-induced NF-kappaB and SAPK activation, blocked TNF-induced GCKR activation. Finally, interference with GCKR expression impeded TRAF2- and TNF-induced SAPK activation but not that of NF-kappaB. This suggests a divergence in the TNF signaling pathway that leads to SAPK and NF-kappaB activation, which is located downstream of TRAF2 but upstream of GCKR. PMID- 9405408 TI - A mammalian homolog of the yeast LCB1 encodes a component of serine palmitoyltransferase, the enzyme catalyzing the first step in sphingolipid synthesis. AB - Serine palmitoyltransferase (SPT; EC 2.3.1.50) catalyzes the initial step dedicated to sphingolipid biosynthesis and is thought to be a key enzyme for regulating cellular sphingolipid content. For SPT activity, the yeast Saccharomyces cerevisiae requires two genes, LCB1 and LCB2. We isolated mammalian LCB1 cDNA homologs from mouse and Chinese hamster ovary (CHO) cells and an LCB2 cDNA homolog from CHO cells. The mammalian LCB1 proteins are predicted to have about 35% amino acid identity to the yeast Lcb1 protein, whereas the CHO LCB2 protein is predicted to have about 40% amino acid identity to the yeast Lcb2 protein. Northern blot analysis of mRNA isolated from various mouse tissues revealed that the tissue distribution of both LCB1 and LCB2 messengers followed a similar pattern. Transfection of an SPT-defective CHO mutant strain with a CHO LCB1-expressing plasmid restored both SPT activity and de novo sphingolipid synthesis to the wild type levels, whereas transfection of the mutant strain with a CHO LCB2-expressing plasmid did not exhibit any recovery effects, indicating that the SPT defect in the mutant cells is specifically complemented by the CHO LCB1 homolog. Furthermore, when the SPT-defective mutant cells were transfected with a plasmid encoding a His6-tagged CHO LCB1 protein, SPT activity bound to a Ni2+-immobilized resin. These results indicate that the CHO LCB1 homolog encodes a component of SPT. PMID- 9405409 TI - Tissue-specific alternative splicing of ascidian troponin I isoforms. Redesign of a protein isoform-generating mechanism during chordate evolution. AB - In vertebrates, troponin I (TnI) exists as shorter and longer isoforms encoded by distinct genes expressed in skeletal and cardiac muscle, respectively. We report that the protochordate ascidian Ciona intestinalis expresses a homologous set of shorter and longer TnI isoforms in body wall muscle and heart, respectively. The heart-specific segment of the ascidian longer TnI isoform shares several sequence features with vertebrate cardiac TnI but lacks the protein kinase A phosphorylation sites implicated in sympatho-adrenal control of cardiac function. In contrast with vertebrates, the ascidian longer and shorter TnI isoforms are produced from a single gene by tissue-specific alternative RNA splicing; remarkably, the molecular mechanism of TnI isoform generation has been entirely reworked during ascidian/vertebrate evolution. Because alternative splicing is the more probable chordate ancestral condition, the long/cardiac versus short/somatic muscle pattern of TnI isoforms likely existed before the occurrence of the gene duplication events that created the vertebrate TnI gene family. Thus, gene duplication was apparently not the primary engine of isoform diversity in this aspect of TnI gene family evolution; rather, it simply provided an alternative (transcriptional) means of maintaining a previously established system of isoform diversity and tissue specificity based on alternative RNA splicing. PMID- 9405410 TI - Purification to homogeneity and reconstitution of the individual components of the epoxide carboxylase multiprotein enzyme complex from Xanthobacter strain Py2. AB - Epoxide metabolism in the aerobic bacterium Xanthobacter strain Py2 proceeds by an NADPH- and NAD+-dependent carboxylation reaction that forms beta-keto acids as products. Epoxide carboxylase, the enzyme catalyzing this reaction, was resolved from the soluble fraction of cell-free extracts into four protein components that are obligately required for functional reconstitution of epoxide carboxylase activity. One of these components, component II, has previously been purified and characterized as an NADPH:disulfide oxidoreductase. In the present study, the three additional epoxide carboxylase components have been purified to homogeneity and characterized. These component proteins are as follows: component I, a homohexameric protein consisting of 41.7-kDa subunits; component III, a dimeric protein consisting of 26.0- and 26.2-kDa polypeptides; and component IV, a dimeric protein consisting of a single 25.4-kDa polypeptide. Component I contained 5 mol of tightly bound zinc per mol of protein. Component I was specifically inactivated by methylepoxypropane, a time-dependent irreversible inactivator of epoxide carboxylase activity, suggesting that this component plays an integral role in epoxide binding and activation. No metals or organic cofactors were detected for components III and IV. The molecular weights, N terminal sequences, and amino acid compositions of the purified epoxide carboxylase components were determined and found to correlate with open reading frames within and adjacent to a cloned fragment of DNA that complements Xanthobacter Py2 mutants defective in epoxide degradation. Using the purified epoxide carboxylase system, epoxide carboxylation was found to be stoichiometrically coupled to the transhydrogenation of pyridine nucleotide cofactors according to the following equation: epoxypropane + CO2 + NADPH + NAD+ -> acetoacetate + H+ + NADP+ + NADH. PMID- 9405411 TI - Targeted disruption of the peripheral-type benzodiazepine receptor gene inhibits steroidogenesis in the R2C Leydig tumor cell line. AB - To evaluate the role of the mitochondrial peripheral-type benzodiazepine receptor (PBR) in steroidogenesis, we developed a molecular approach based on the disruption of the PBR gene, by homologous recombination, in the constitutive steroid producing R2C rat Leydig tumor cell line. Inactivation of one allele of the PBR gene resulted in the suppression of PBR mRNA and ligand binding expression. Immunoblot and electron microscopic immunogold labeling analyses confirmed the absence of the 18-kDa PBR protein in the selected clone. Although mitochondria from the PBR-negative cells contained high levels of the constitutively expressed 30-kDa steroidogenic activity regulator protein, these cells produced minimal amounts of steroids compared with normal cells (5%). Moreover, mitochondria from PBR-negative cells failed to produce pregnenolone when supplied with exogenous cholesterol. Addition of the hydrosoluble cholesterol derivative, 22R-hydroxycholesterol, increased steroid production by the PBR-negative R2C cells, indicating that the cholesterol transport mechanism was impaired. Stable transfection of the PBR-negative R2C Leydig cells with a vector containing the PBR cDNA resulted in the recovery of the steroidogenic function of the cells. These data demonstrate that PBR is an indispensable element of the steroidogenic machinery, where it mediates the delivery of the substrate cholesterol to the inner mitochondrial side chain cleavage cytochrome P 450. PMID- 9405412 TI - Depression of T-cell epitope generation by stabilizing hen lysozyme. AB - Conformational stability of proteins is an important factor that determines their resistance/susceptibility to proteolytic digestion. Intracellular proteolysis is the key step in antigen presentation events for protein antigens; hence, it is likely that increasing protein stability reduces the antigenicity of proteins. We prepared three hen egg white lysozyme derivatives possessing different stabilities by chemical modification to clarify the relationship between conformational stability and the antigenicity of the protein. One of the derivatives was conformationally unstabilized by removing one intramolecular disulfide bond, whereas the two others were stabilized by the addition of an intramolecular cross-link. The antigenicity of these derivatives was evaluated using hen egg white lysozyme-specific T-cell hybridoma cells and a B-lymphoma cell line, A20, as antigen-presenting cells. With an increase in conformational stability, the T-cell response decreased. However, the reduction was not derived from the inefficiency of internalization to A20 cells nor the alteration of antigenicity by chemical modifications. Moreover, from analyses of their susceptibility to proteolysis and the kinetics of presentation of the T-cell epitope, it was confirmed that increasing protein stability led to the depression of T-cell epitope generation by increasing resistance to proteolysis. These results have an important implication in devising a new strategy for manipulating T-cell response by the stability of protein antigen. PMID- 9405413 TI - Replacement of conserved cysteines in human tissue inhibitor of metalloproteinases-1. AB - Tissue inhibitor of metalloproteinases-1 (TIMP-1) is resistant to extremes of temperature and pH. This is thought to be due in part to the presence of six sulfhydryl bridges presumed to maintain the structural integrity of the molecule. As part of a study looking at structure-function relationships, a number of the conserved cysteine residues in TIMP-1 were targeted for replacement with serine. Single and double replacements of these conserved cysteines, as well as replacements around these cysteines, were expressed using a vaccinia virus system and analyzed for functional and structural competence. Analysis by circular dichroism indicated that these mutants maintained secondary structures similar to those of wild-type TIMP-1. Trypsin susceptibility experiments indicated that the tertiary structure of the mutants had not been drastically changed. Analysis of functional competence demonstrated that there were significant changes in some of these mutants. Assays using collagen fibrils or gelatin as substrates indicated that the double mutant C1S/C70S, but not C3S/C99S, had lost inhibitory activity against human fibroblast-type collagenase (FIB-CL) and at high concentrations only had slight activity against Mr 72,000 gelatinase (Mr 72,000 gelatinase). Kinetic analysis of TIMP-1 inhibition of FIB-CL cleavage of a peptide substrate indicated that mutants C1S/C70S, C3S/C99S, and CEEC --> CQQC retained their ability to inhibit FIB-CL in a manner similar to wild-type TIMP-1, while mutants C1S and C70S showed little inhibitory activity. The mutants C99S and C137S could also inhibit FIB-CL cleavage of the peptide substrate. The results indicated that the degree of inhibition by the TIMP-1 mutants varied somewhat depending on the choice of substrates. Interestingly, replacing both cysteines from a disulfide bond in the wild-type molecule resulted in a more competent inhibitor than either of the single site "parent" mutations. Taken together, these experiments indicate that TIMP-1 can be rendered inactive by the loss of a single cysteine. PMID- 9405414 TI - Mutations in a bacterial mechanosensitive channel change the cellular response to osmotic stress. AB - MscL is a channel found in bacterial plasma membranes that opens a large pore in response to mechanical stress. Here we demonstrate that some mutations within this channel protein (K31D and K31E) evoke a cellular phenotype in which the growth rate is severely depressed. Increasing the osmolarity of the growth medium partially rescues this "slowed growth" phenotype and decreases an abnormal cytosolic potassium loss observed in cells expressing the mutants. In addition, upon sudden decrease in osmolarity (osmotic downshock) more cytoplasmic potassium is released from cells expressing the mutants than cells expressing wild-type MscL. After osmotic downshock, all cells remained viable; hence, the differences in potassium efflux observed are not due to cell lysis but instead appear to be an exaggeration of the normal response to this sudden change in environmental osmolarity. Patch clamp studies in native bacterial membranes substantiate the hypothesis that these mutant channels are more sensitive to mechanical stresses, especially at voltages approaching those estimated for bacterial membrane potentials. These data are consistent with a crucial role for MscL in the adaptation to large osmotic downshock and suggest that if the normally tight regulation of MscL gating is disrupted, cell growth can be severely inhibited. PMID- 9405415 TI - Calorimetric observation of a GroEL-protein binding reaction with little contribution of hydrophobic interaction. AB - Binding of Escherichia coli chaperonin, GroEL, to substrate proteins with non native structure, reduced alpha-lactalbumin (rLA) and denatured pepsin, were analyzed by isothermal titration calorimetry at various temperatures in the presence of salt (0.2 M KCl). Both proteins bound to GroEL with 1:1 stoichiometry and micromolar affinity at all temperatures tested. However, thermodynamic properties of their binding to GroEL are remarkably different from each other. While heat capacity changes (DeltaCp) of rLA-GroEL binding showed large negative values, -4.19 kJ mol-1 K-1, that of denatured pepsin-GroEL binding was only -0.2 kJ mol-1 K-1. These values strongly indicate that the hydrophobic interaction is a major force of rLA-GroEL binding but not so for denatured pepsin-GroEL binding. When salt was omitted from the solution, the affinity and DeltaCp of the rLA GroEL binding reaction were not significantly changed whereas denatured pepsin lost affinity to GroEL. Thus, in the non-native protein-GroEL binding reaction, thermodynamic properties, as well as the effect of salt, differ from protein to protein and hydrophobic interaction may not always be a major driving force. PMID- 9405416 TI - c-Jun NH2-terminal kinases target the ubiquitination of their associated transcription factors. AB - Regulatory proteins are often ubiquitinated, depending on their phosphorylation status as well as on their association with ancillary proteins that serve as adapters of the ubiquitination machinery. We previously demonstrated that c-Jun is targeted for ubiquitination by its association with inactive c-Jun NH2 terminal kinase (JNK). Phosphorylation by activated JNK protects c-Jun from ubiquitination, thus by prolonging its half-life. In the study reported here, we determined the ability of JNK to target ubiquitination of its other substrates (Elk1 and activating transcription factor 2 (ATF2)) and associated proteins (ATF2 and JunB). We demonstrate that phosphorylation by JNK protects ATF2, but not Elk1, from JNK-targeted ubiquitination. We also show that association of inactive JNK with JunB or ATF2 is necessary to target them for ubiquitination. Unlike its targeting of c-Jun, JNK requires additional cellular components, yet to be identified, to target the ubiquitination of ATF2. Elk1 is phosphorylated by JNK, but JNK neither associates with nor targets Elk1 for ubiquitination. The implications for the dual role of JNK in the regulation of ubiquitination and stability of c-Jun, ATF2, and JunB in normally growing versus stressed cells are discussed. PMID- 9405417 TI - Inhibition of inducible nitric oxide synthase expression by interferons alpha and beta in bovine retinal pigmented epithelial cells. AB - Bovine retinal pigmented epithelial (RPE) cells express an inducible nitric oxide synthase (NOS-2) after activation with interferon (IFN)-gamma and lipopolysaccharide (LPS). Experiments were performed to investigate the effects of IFN-alpha and IFN-beta on NOS-2 activity. These types of interferons did not aid LPS in the production of nitrite, but markedly inhibited in a concentration dependent manner the nitrite release due to LPS/IFN-gamma. Analysis by Western and Northern blots showed that RPE cells co-stimulated with IFN-alpha or IFN-beta with LPS/IFN-gamma accumulated lower levels of NOS-2 protein and mRNA than in the presence of LPS/IFN-gamma alone. The presence of IFN-alpha or IFN-beta did not accelerate mRNA degradation, implying that these interferons did not affect NOS-2 mRNA stability, but more probably NOS-2 gene expression. Furthermore, IFN-gamma binding studies demonstrated that the inhibitory effect of IFN-alpha and IFN-beta is not caused by a blocking of IFN-gamma receptors. Analysis of NF-kappaB activation by electrophoretic mobility shift assay demonstrated that LPS/IFN gamma-induced NF-kappaB binding was not changed by the presence of IFN-alpha. However, similar experiments revealed that the activation of interferon regulatory factor-1 (IRF-1) by LPS/IFN-gamma was decreased by IFN-alpha. This phenomenon could be due to the decline of IRF-1 mRNA and the up-regulation of IRF 2 mRNA, an IRF-1 repressor, by IFN-alpha. These results suggest that the inhibitory effect of IFN-alpha and -beta on NOS-2 induction could be partially explained by their effect on the induction of the IRFs, which were involved in NOS-2 gene transcription. PMID- 9405418 TI - Specific, high affinity binding sites for an antifungal plant defensin on Neurospora crassa hyphae and microsomal membranes. AB - Hs-AFP1, an antifungal plant defensin from seed of the plant Heuchera sanguinea, was radioactively labeled using t-butoxycarbonyl-[35S]L-methionine N hydroxysuccinimidyl ester, resulting in a 35S-labeled peptide with unaltered antifungal activity. [35S]Hs-AFP1 was used to assess binding on living hyphae of the fungus Neurospora crassa. Binding of [35S]Hs-AFP1 was found to be competitive, reversible, and saturable with an apparent Kd of 29 nM and a Bmax of 1.4 pmol/mg protein. [35S]Hs-AFP1 also bound specifically and reversibly to microsomal membranes derived from N. crassa hyphae with a Kd of 27 nM and a Bmax of 102 pmol/mg protein. The similarity in Kd value between binding sites on hyphae and microsomes indicates that Hs-AFP1 binding sites reside on the plasma membrane. Binding of [35S]Hs-AFP1 to both hyphae and microsomal membranes could be competed to some extent by four different structurally related plant defensins but not by various structurally unrelated antimicrobial peptides. In addition, an inactive single amino acid substitution variant of the antifungal plant defensin Rs-AFP2 from Raphanus sativus seed was also unable to displace [35S]Hs-AFP1 from its binding sites, whereas Rs-AFP2 itself was able to compete with [35S]Hs-AFP1. PMID- 9405419 TI - The structure and function of the actin-binding domain of myosin light chain kinase of smooth muscle. AB - In addition to its kinase activity, the myosin light chain kinase (MLCK) of smooth muscle has an actin binding activity through which it can regulate the actin-myosin interaction of smooth muscle (Kohama, K., Okagaki, T., Hayakawa, K., Lin, Y., Ishikawa, R., Shimmen, T., and Inoue, A. (1992) Biochem. Biophys. Res. Commun. 184, 1204-1211). In this study, we have analyzed the actin binding activity of MLCK and related it to its amino acid sequence by producing native and recombinant fragments of MLCK. Parent MLCK exhibited both calcium ion (Ca2+) and calmodulin (Ca2+/CaM)-sensitive and Ca2+/CaM-insensitive binding to actin filaments. The native fragment, which consists of the Met1-Lys114 sequence (Kanoh, S., Ito, M., Niwa, E., Kawano, Y., and Hartshorne, D. J. (1993) Biochemistry 32, 8902-8907), and the recombinant NN fragment, which contains this 1-114 sequence, showed only Ca2+/CaM-sensitive binding. An inhibitory effect of the NN fragment on the actin-myosin interaction was observed by assaying in vitro motility and by measuring the actin-activated ATPase activity of myosin. The recombinant NN/41 fragment, which is constructed without the Met1-Pro41 sequence of the NN fragment, lost both the actin binding activity and the inhibitory effect. We confirmed the importance of the 1-41 sequence by using a few synthetic peptides to compete against the NN fragment in binding to actin filaments. The experiments using recombinant fragments and synthetic peptides also revealed that the site for CaM-binding is the Pro26-Pro41 sequence. The site for the Ca2+/CaM insensitive binding, which is shown to be localized between the Ca2+/CaM sensitive site and the central kinase domain of MLCK, exerted no regulatory effects on the actin-myosin interaction. PMID- 9405421 TI - Identification of cell binding sequences in mouse laminin gamma1 chain by systematic peptide screening. AB - Laminin-1, a major component of basement membranes, consists of three different chains designated alpha1, beta1, and gamma1 and has diverse biological functions. We have identified cell binding sites on the mouse laminin gamma1 chain, using systematic screening of 165 overlapping synthetic peptides covering the entire chain. We identified 12 cell binding sequences using HT-1080 human fibrosarcoma and B16-F10 mouse melanoma cells in two independent assays employing peptide conjugated Sepharose beads and peptide-coated dishes. Four peptides (C-16, C-28, C-64, and C-68) located on the globular domains of the gamma1 chain were the most active and showed dose-dependent cell attachment. Cell attachment to C-68 was inhibited by EDTA and by anti-alpha2beta1 integrin antibodies. Cell attachment to C-16 and C-64 was partially inhibited by EDTA but was not inhibited by anti integrin antibodies. EDTA and anti-integrin antibodies did not affect cell attachment to C-28. The four peptides were tested in adhesion and differentiation assays with endothelial, neuronal, and human salivary gland cells. C-16 was the most active for all of the cells, whereas the other three peptides showed cell type specificity in their activities. The active core sequences of C-16, C-28, C 64, and C-68 are YVRL, IRVTLN, TTVKYIFR, and SIKIRGTY, respectively. These sequences are highly conserved among the different species and in the laminin gamma2 chain. These results suggest that the specific sequences on the laminin gamma1 chain are biologically active and interact with distinct cell surface receptors. PMID- 9405420 TI - Conformational integrity and ligand binding properties of a single chain T-cell receptor expressed in Escherichia coli. AB - We recently showed that a soluble, heterodimeric murine D10 T-cell receptor (TCR) (Valpha2Calpha, Vbeta8.2Cbeta) expressed in insect cells binds both Vbeta8.2 specific bacterial superantigen staphylococcal enterotoxin C2 (SEC2) and a soluble, heterodimeric major histocompatibility complex class II I-Ak.conalbumin peptide complex with a low micromolar affinity. To define further the structural requirements for the TCR/ligand interactions, we have produced in Escherichia coli a soluble, functional D10 single chain (sc) TCR molecule in which the Valpha and Vbeta domains are connected by a flexible peptide linker. Purified and refolded D10 scTCR bound to SEC2 and murine major histocompatibility complex class II I-Ak.conalbumin peptide complex with thermodynamic and kinetic binding constants similar to those measured for the baculovirus-derived heterodimeric D10 TCR suggesting that neither the TCR constant domains nor potential N- or O-linked carbohydrate moieties are necessary for ligand recognition and for expression and proper folding of the D10 scTCR. Purified D10 scTCR remained soluble at concentrations up to 1 mM. Circular dichroism and NMR spectroscopy indicated that D10 scTCR is stabilized predominantly by beta-sheet secondary structure, consistent with its native-like conformation. Because of its limited size, high solubility, and structural integrity, purified D10 scTCR appears to be suitable for structural studies by multidimensional NMR spectroscopy. PMID- 9405422 TI - Renaturation of rhodanese by translational elongation factor (EF) Tu. Protein refolding by EF-Tu flexing. AB - The translation elongation factor (EF) Tu has chaperone-like capacity to promote renaturation of denatured rhodanese. This renaturation activity is greatly increased under conditions in which the factor can oscillate between the open and closed conformations that are induced by GDP and GTP, respectively. Oscillation occurs during GTP hydrolysis and subsequent replacement of GDP by EF-Ts which is then displaced by GTP. Renaturation of rhodanese and GTP hydrolysis by EF-Tu are greatly enhanced by the guanine nucleotide exchange factor EF-Ts. However, renaturation is reduced under conditions that stabilize EF-Tu in either the open or closed conformation. Both GDP and the nonhydrolyzable analog of GTP, GMP-PCP, inhibit renaturation. Kirromycin and pulvomycin, antibiotics that specifically bind to EF-Tu and inhibit its activity in peptide elongation, also strongly inhibit EF-Tu-mediated renaturation of denatured rhodanese to levels near those observed for spontaneous, unassisted refolding. Kirromycin locks EF-Tu in the open conformation in the presence of either GTP or GDP, whereas pulvomycin locks the factor in the closed conformation. The results lead to the conclusion that flexing of EF-Tu, especially as occurs between its open and closed conformations, is a major factor in its chaperone-like refolding activity. PMID- 9405423 TI - Nucleotide-depleted beef heart F1-ATPase exhibits strong positive catalytic cooperativity. AB - Catalytic cooperativity is a central feature of the binding change mechanism for F0F1-ATP synthases. However, in a recent publication (Reynafarje, B. D., and Pedersen, P. L. (1996) J. Biol. Chem. 271, 32546-32550), Reynafarje and Pedersen claim that cooperative effects are an artifact caused by endogenous nucleotides and that when such nucleotides are removed, the multiple catalytic sites on MF1 behave independently during ATP hydrolysis. In contrast to this conclusion, we show here that when ATP is loaded at a single catalytic site on nucleotide depleted MF1, the rate of product release is accelerated by up to 5 x 10(4)-fold by the binding of ATP at adjacent catalytic sites. Hence, nucleotide-depleted MF1 is not an exception but does in fact show strong cooperative interactions. In addition, evidence is presented supporting a random order for product release during ATP hydrolysis. PMID- 9405424 TI - Identification of modification sites in large biomolecules by stable isotope labeling and tandem high resolution mass spectrometry. The active site nucleophile of thiaminase I. AB - A widely used procedure for site localization of covalent protein modifications involves proteolysis, partial chromatographic separation of the resulting complex mixture, and tandem mass spectrometry (MS/MS) to identify peptides whose molecular weight (Mr) has been increased appropriately by the modification. As found previously for MS of small molecules, this study shows that protein fragment identification can be greatly simplified by labeling the modification with stable isotopes. Further, the high resolution capabilities of Fourier transform MS make possible the direct identification of CH3/CD3-labeled peptides without chromatographic separation. Although separate Asp-N, Lys-C, and alpha chymotrypsin digests of thiaminase I (42 kDa) yielded as many as 70 peptides, FTMS identification of the labeled peptide localized the modification site of a mechanism-based inhibitor to Arg101-Lys121, Asp90-Gly122, and Gly107-Tyr119, respectively. The measured mass difference values of the two labels agreed with that expected for CH3/CD3, 3.019 Da, with a standard deviation of 0.005 Da, providing persuasive identity verification. MS/MS fragmentation narrowed the site to Pro109-Phe118 and also caused loss of the derivative with a sulfur atom, uniquely identifying Cys113 as the thiaminase I active-site nucleophile among the 379 amino acids. PMID- 9405425 TI - Separate domains of the human fas ligand dictate self-association and receptor binding. AB - The Fas receptor rapidly induces apoptosis when activated by ligand binding or by cross-linking with anti-Fas antibody. The Fas ligand (FasL), a member of the tumor necrosis factor family of ligands, is a 40-kilodalton type II transmembrane protein which is cleaved to produce soluble ligand. Although the Fas-FasL interaction plays a critical role in peripheral T cell homeostasis and cytotoxic T lymphocyte-mediated target cell killing, the requirements for human FasL receptor binding and oligomerization have not been defined. Here we report two distinct domains of the ligand which are responsible for self-association and binding to the Fas receptor. A COOH-terminal sequence of the FasL was found to be required for binding and biological activity, as verified by deletion mutagenesis, use of the NOK-1 blocking antibody and the humanized gld FasL mutation. N-Linked glycosylation of the FasL was not required for biological activity. However, the FasL expression level was dependent upon the three N linked glycosylation sites. Moreover, the ability of the FasL to self-associate was not dependent upon transmembrane or cytoplasmic sequences, but was localized to a 47-amino acid region in its extracellular domain. These results indicate that the FasL-Fas receptor complex depends upon independent motifs located within the extracellular domain of the FasL. PMID- 9405426 TI - Characterization of Escherichia coli endonuclease VIII. AB - Escherichia coli endonuclease VIII (endo VIII) was identified as an enzyme that, like endonuclease III (endo III), removes radiolysis products of thymine including thymine glycol, dihydrothymine, beta-ureidoisobutyric acid, and urea from double-stranded plasmid or phage DNA and cleaves the DNA strand at abasic (AP) sites (Melamede, R. J., Hatahet, Z., Kow, Y. W., Ide., H., and Wallace, S. S. (1994) Biochemistry 33, 1255-1264). Using apparently homogeneous endo VIII protein, we now show that endo VIII removes from double-stranded oligodeoxyribonucleotides the stable oxidative products of cytosine, 5 hydroxycytosine and 5-hydroxyuracil. Endo VIII cleaved the damage-containing DNA strand by beta,delta-elimination as does formamidopyrimidine DNA glycosylase (Fpg). Like Fpg, endo VIII also excised the 5'-terminal deoxyribose phosphate from an endonuclease IV (endo IV) pre-incised AP site. Thus, in addition to amino acid sequence homology (Jiang, D., Hatahet, Z., Blaisdell, J., Melamede, R. J., and Wallace, S. S. (1997) J. Bacteriol. 179, 3773-3782), endo VIII shares a number of catalytic properties with Fpg. In addition, endo VIII specifically bound to oligodeoxynucleotides containing a reduced AP site with a stoichiometry of 1:1 for protein to DNA with an apparent equilibrium dissociation constant of 3.9 nM. Like Fpg and endo III, the DNase I footprint was small with contact sites primarily on the damage-containing strand; unlike Fpg and endo III, the DNA binding of endo VIII to DNA was asymmetric, 3' to the reduced AP site. PMID- 9405427 TI - Farnesol inhibits L-type Ca2+ channels in vascular smooth muscle cells. AB - Earlier experiments with animal and human arteries have shown that farnesol, a natural 15-carbon (C15) isoprenoid, is an inhibitor of vasoconstriction (Roullet, J.-B., Xue, H., Chapman, J., McDougal, P., Roullet, C. M., and McCarron, D. A. (1996) J. Clin. Invest. 97, 2384-2390). We report here that farnesol reduced KCl- and norepinephrine-dependent cytosolic Ca2+ transients in fura-2-loaded intact arteries. An effect on Ca2+ signaling was also observed in cultured aortic smooth muscle cells (A10 cells). In these cells, farnesol reduced KCl-induced [Ca2+]i transients and mimicked the inhibitory effect of Ca2+-free medium on the [Ca2+]i response to both 12,13-phorbol myristate acetate, a protein kinase C activator, and thapsigargin, a specific endoplasmic reticulum ATPase inhibitor. Perforated patch-clamp experiments further showed in two vascular smooth muscle cell lines (A10 and A7r5), a reversible, dose-dependent inhibitory effect of farnesol on L type Ca2+ currents (IC50 = 2.2 microM). Shorter (C10, geraniol) and longer (C20, geranylgeraniol) isoprenols were inactive. L-type Ca2+ channel blockade also occurred under tight (gigaohm) seal configuration using cell-attached, single channel analysis, thus suggesting a possible action of farnesol from within the intracellular space. We finally demonstrated that farnesol did not affect Ca2+ sensitive pathways implicated in smooth muscle contraction, as tested with alpha toxin permeabilized arteries. Altogether, our results indicate that farnesol is an inhibitor of vascular smooth muscle Ca2+ signaling with plasma membrane Ca2+ channel blocker properties. The data have implications for the endogenous and pharmacological regulation of vascular tone by farnesol or farnesol analogues. PMID- 9405428 TI - Differential effects of the swedish mutant amyloid precursor protein on beta amyloid accumulation and secretion in neurons and nonneuronal cells. AB - Expression of the Swedish DeltaNL mutation in the beta-amyloid precursor protein (APPDeltaNL) dramatically increases Abeta generation in nonneuronal cell lines, although it is unclear whether intracellular levels of beta-amyloid (Abeta) are also elevated after APPDeltaNL expression. Furthermore, the effects of expressing APPDeltaNL in neurons on the production and secretion of Abeta-(1-40) and Abeta (1-42) are unknown. To address these issues, we examined the generation of both intracellular and secreted Abeta-(1-40) and Abeta-(1-42) in human neuronal NT2N cells, in primary rat astrocytes, and in Chinese hamster ovary cells engineered to express wild-type APP or APPDeltaNL using a recombinant Semliki Forest virus expression system. Expression of APPDeltaNL led to a marked increase in APPbeta and the C-terminal fragment containing the entire Abeta sequence (C99) in all cells tested. However, a dramatic elevation of intracellular and secreted Abeta (1-40) and Abeta-(1-42) was seen only in astrocytes and Chinese hamster ovary cells. The DeltaNL mutation did not cause a significant increase in intracellular or secreted Abeta-(1-40) or Abeta-(1-42) in NT2N cells. Since NT2N cells expressing APPDeltaNL accumulate much higher levels of C99 than cells expressing wild-type APP, we conclude that the rate-limiting step in Abeta production could be the further processing of C99 by gamma-secretase in these cells. These results show that the Swedish DeltaNL mutation causes nonneuronal cells to process APP via pathways more in common with the metabolism of wild-type APP in neurons. PMID- 9405429 TI - The gene encoding the elongation factor P protein is essential for viability and is required for protein synthesis. AB - Elongation factor P (EFP) is a protein that stimulates the peptidyltransferase activity of fully assembled 70 S prokaryotic ribosomes and enhances the synthesis of certain dipeptides initiated by N-formylmethionine. This reaction appears conserved throughout species and is promoted in eukaryotic cells by a homologous protein, eIF5A. Here we ask whether the Escherichia coli gene encoding EFP is essential for cell viability. A kanamycin resistance (KanR) gene was inserted near the N-terminal end of the efp gene and was cloned into a plasmid, pMAK705, that has a temperature-sensitive origin of replication. After transformation into a recA+ E. coli strain, temperature-sensitive mutants were isolated, and their chromosomal DNA was sequenced. Mutants containing the efp-KanR gene in the chromosome grew at 33 degrees C only in the presence of the wild-type copy of the efp gene in the pMAK705 plasmid and were unable to grow at 44 degrees C. Incorporation of various isotopes in vivo suggests that translation is impaired in the efp mutant at 44 degrees C. At 44 degrees C, mutant cells are severely defective in peptide-bond formation. We conclude that the efp gene is essential for cell viability and is required for protein synthesis. PMID- 9405430 TI - Role of estrogen receptor gene demethylation and DNA methyltransferase.DNA adduct formation in 5-aza-2'deoxycytidine-induced cytotoxicity in human breast cancer cells. AB - The cytosine analog 5-aza-2'-deoxycytidine is a potent inhibitor of DNA methyltransferase. Its cytotoxicity has been attributed to several possible mechanisms including reexpression of growth suppressor genes and formation of covalent adducts between DNA methyltransferase and 5-aza-2'-deoxycytidine substituted DNA which may lead to steric inhibition of DNA function. In this study, we use a panel of human breast cancer cell lines as a model system to examine the relative contribution of two mechanisms, gene reactivation and adduct formation. Estrogen receptor-negative cells, which have a hypermethylated estrogen receptor gene promoter, are more sensitive than estrogen receptor positive cells and underwent apoptosis in response to 5-aza-2'-deoxycytidine. For the first time, we show that reactivation of a gene silenced by methylation, estrogen receptor, plays a major role in this toxicity in one estrogen receptor negative cell line as treatment of the cells with anti-estrogen-blocked cell death. However, drug sensitivity of other tumor cell lines correlated best with increased levels of DNA methyltransferase activity and formation DNA.DNA methyltransferase adducts as analyzed in situ. Therefore, both reexpression of genes like estrogen receptor and formation of covalent enzyme. DNA adducts can play a role in 5-aza-2'-deoxycytidine toxicity in cancer cells. PMID- 9405431 TI - Asymmetric interactions of hexameric bacteriophage T7 DNA helicase with the 5'- and 3'-tails of the forked DNA substrate. AB - Bacteriophage T7 DNA helicase requires two noncomplementary single-stranded DNA (ssDNA) tails next to a double-stranded DNA (dsDNA) region to initiate DNA unwinding. The interactions of the helicase with the DNA were investigated using a series of forked DNAs. Our results show that the helicase interacts asymmetrically with the two tails of the forked DNA. When the helicase was preassembled on the forked DNA before the start of unwinding, a DNA with 15 nucleotide (nt) 3'-tail and 35-nt 5'-tail was unwound with optimal rates close to 60 base pairs/s at 18 degrees C. When the helicase was not preassembled on the DNA, a >65-nt long 5'-tail was required for maximal unwinding rates of 12 base pairs/s. We show that the helicase interacts specifically with the ssDNA region and maintains contact with both ssDNA strands during DNA unwinding, since conversion of the two ssDNA tails to dsDNA structures greatly inhibited unwinding, and the helicase was unable to unwind past a nick in the dsDNA region. These studies have provided new insights into the mechanism of DNA unwinding. We propose an exclusion model of DNA unwinding in which T7 helicase hexamer interacts mainly with the ssDNA strands during DNA unwinding, encircling the 5' strand and excluding the 3'-strand from the hole. PMID- 9405432 TI - Hyperosmolarity-induced gene stimulation is mediated by the negative calcium responsive element. AB - The negative calcium responsive elements of the parathyroid hormone gene bind to a specific set of nuclear proteins in an extracellular calcium (Ca2+e)-dependent manner. We have found that one of the negative calcium responsive elements, named oligo B, is found in the 5'-flanking region of such vasoactive genes as the vasopressin and atrial natriuretic polypeptide genes. Furthermore, the oligo B like sequence in the former gene is conserved throughout evolution. Because expression of some of these vasoactive genes is altered by external stimuli which change cell volume, we examined whether oligo B is involved in gene regulation by hyperosmolarity. Here, we demonstrate that the binding between oligo B and its binding nuclear proteins including a redox factor 1 was reduced by hyperosmolarity generated by sodium chloride but not by urea. Such attenuated binding was reversed by dephosphorylating nuclear proteins by a potato acid phosphatase, suggesting that NaCl treatment elicited phosphorylation of these nuclear proteins to weaken their binding activity to oligo B. Furthermore, these nuclear events led to hyperosmolarity-mediated transcriptional stimulation of the genes bearing this DNA element in the cultured cells. PMID- 9405433 TI - A ubiquitin-specific protease that efficiently cleaves the ubiquitin-proline bond. AB - Ubiquitin is a small eukaryotic protein that is synthesized naturally as one of several fusion proteins, which are processed by ubiquitin-specific proteases to release free ubiquitin. The expression of heterologous proteins as fusions to ubiquitin in either prokaryotic or eukaryotic hosts often dramatically enhances their yield, and allows the exposure of any amino acid following cleavage of ubiquitin. The single exception is when proline is the amino acid immediately following ubiquitin; the ubiquitin-proline bond is poorly cleaved by presently studied ubiquitin-specific proteases. We show that the mouse ubiquitin-specific protease Unp, and its human homolog Unph, can efficiently cleave the ubiquitin proline bond in ubiquitin fusion proteins of different sizes. N-terminal sequencing of the cleavage products reveals that cleavage occurs precisely at the ubiquitin-proline junction. The biological significance of this cleavage activity is unclear, as ubiquitin-proline fusions do not occur naturally. However, it may indicate a different catalytic mechanism for these ubiquitin-specific proteases and/or that they can cleave ubiquitin-like proteins. Unp and Unph thus represent versatile ubiquitin-specific proteases for cleaving ubiquitin-fusion proteins in biotechnology and basic research, regardless of both the amino acid immediately following ubiquitin, and the size of the fusion partner. PMID- 9405434 TI - Redundancy of class III POU proteins in the oligodendrocyte lineage. AB - Class III POU proteins are prominent regulators of neural development. Tst 1/Oct6/SCIP, for instance, is essential for terminal differentiation of myelinating Schwann cells in the peripheral nervous system. Although Tst 1/Oct6/SCIP is also expressed in the myelin forming oligodendrocytes of the central nervous system, targeted deletion of Tst-1/Oct6/SCIP failed to reveal a gross alteration of myelination in the central nervous system. To better understand this apparent discrepancy, we examined the expression of POU proteins in both cultured primary oligodendrocytes and in the oligodendrocyte-like CG-4 cell line. These cells expressed Tst-1/Oct6/SCIP, Brn-1, and Brn-2 in significant amounts, indicating that Brn-1 and Brn-2 might have the capacity to compensate loss of Tst-1/Oct6/SCIP. We show that Tst-1/Oct6/SCIP, Brn-1, and Brn-2 were all down-regulated during the early phases of oligodendrocyte development both on RNA and protein level. All three POU proteins exhibited similar DNA binding characteristics. When promoters consisting of a single POU protein-binding site adjacent to a TATA box were used as reporters in transient transfections, Brn-1 proved to be a weaker transcriptional activator than Tst-1/Oct6/SCIP. In agreement with this, we found the transactivation domain of Brn-1, which we mapped between amino acids 119 and 237, significantly weaker than the transactivation domain of Tst-1/Oct6/SCIP. Taken together, our data imply a partial, but not complete redundancy between POU proteins in oligodendrocytes. PMID- 9405435 TI - Photoinactivation of the F1-ATPase from spinach chloroplasts by dequalinium is accompanied by derivatization of methionine beta183. AB - In contrast to the F1-ATPases from bovine mitochondria and the thermophilic Bacillus PS3, which are reversibly inhibited by dequalinium in the absence of irradiation, the Mg2+-ATPase activity of heat- or dithiothreitol-activated chloroplast F1 (CF1) from spinach chloroplasts is slightly stimulated by dequalinium. Conversely, dequalinium is a partial inhibitor (maximal inhibition is 85-90%) of the Ca2+-ATPase of CF1 activated by heat, dithiothreitol, or octylglucoside. The Mg2+- and Ca2+-ATPase activities of CF1 respond differently in the presence of lauryl dimethylamine oxide (LDAO) in the assay medium. Whereas the Mg2+-ATPase activity of heat- or dithiothreitol-activated CF1 is stimulated up to 14-fold by increasing concentrations of LDAO, the Ca2+-ATPase is inhibited in a biphasic manner by increasing concentrations of LDAO. In the presence of LDAO, dequalinium does not stimulate the heat-activated Mg2+-ATPase over that promoted by LDAO alone. That dequalinium slightly stimulates Mg2+-ATPase activity although it inhibits Ca2+-ATPase activity can be reconciled by assuming that dequalinium binds to two sites in CF1, a stimulatory site that also binds LDAO and an inhibitory site. By acting as a partial inhibitor of the Mg2+-ATPase activity that it activates, the combined effect of dequalinium is modest stimulation. Irradiation of heat- or dithiothreitol-activated CF1 or the alpha3beta3gamma subcomplex of CF1 in the presence of 12 microM dequalinium led to rapid photoinactivation. ATP and ADP, separately or in combination with Mg2+, protect against photoinactivation. After photoinactivating the alpha3beta3gamma subcomplex of CF1 with [14C]dequalinium, tryptic and peptic digests of the isolated, derivatized beta subunit were fractionated by high performance liquid chromatography. Sequencing of the isolated, radioactive tryptic and peptic peptides revealed that Metbeta183, which is at or near the catalytic site, is derivatized in a single beta subunit when CF1 is photoinactivated with [14C]dequalinium. PMID- 9405436 TI - Promoter activation via a cyclic AMP response element in vitro. AB - Transcription activation via activating transcription factor cyclic AMP response element binding (ATF/CREB) sites in vitro was explored using transcription and permanganate assay for open complex formation. These sites were used to drive transcription from an adenovirus major late core sequence. Under conditions where activation is strong, 20-50-fold, ATF/CREB is required for preinitiation complexes to reach the open complex stage. Complete opening requires activator, ATP, and initiating nucleotides. In exploration of postinitiation steps, no stimulation of promoter clearance was observed but a modest stimulation of the rate of continuous transcription occurred. High amounts of DNA template, commonly used in in vitro studies, allows some templates to open without activator, but leaves the nucleotide requirements intact. This leads to a drastic lowering of the dependence on ATF/CREB. Taken together, the data indicate that ATF/CREB activates this system primarily by stimulating the formation of functional preinitiation complexes. PMID- 9405437 TI - Multisite phosphorylation and the nuclear localization of phosphatase inhibitor 2 green fluorescent protein fusion protein during S phase of the cell growth cycle. AB - Human phosphatase inhibitor 2 (Inh2) is a phosphoprotein that complexes with type 1 protein phosphatase, and its expression peaks during S phase and mitosis during the cell cycle. Localization of Inh2 was visualized in HS68 human fibroblasts by fusing Inh2 to green fluorescent protein (GFP). During G1 phase, Inh2-GFP was localized in the cytoplasm, and as cells progressed into S phase Inh2-GFP accumulated in the nucleus. Known phosphorylation sites of Inh2 at Thr-72, Ser 86, and Ser-120/121 were each replaced with alanine. None of the mutated Inh2-GFP proteins accumulated in the nucleus during S phase, indicating that all of these phosphorylation sites were required. Mutation of two lysine residues in a putative nuclear localization sequence in Inh2 also prevented the Inh2-GFP fusion protein from accumulating in the nucleus during S phase. Recombinant Inh2 was phosphorylated by kinases in cytosols prepared from G1 and S phase cells. The amount of Inh2 kinase attributed to casein kinase 2, based on inhibition by heparin, increased 2.6-fold from G1 to S phase. In addition, kinases in G1 versus S phase cytosols produced distinct Inh2 phosphopeptides. The results indicate that changes in phosphorylation of Inh2 are involved in intracellular redistribution of the protein during the cell cycle. PMID- 9405438 TI - Protection against oxidative stress-induced cell death by intracellular platelet activating factor-acetylhydrolase II. AB - Platelet-activating factor-acetylhydrolase (PAF-AH), which removes the acetyl group at the sn-2 position of PAF, is distributed widely in tissues and plasma. Tissue cytosol contains at least two types of PAF-AH, isoforms Ib and II. Isoform Ib is a tertiary G-protein complex-like heterotrimeric enzyme that is involved in brain development such as formation of the brain cortex. Isoform II (PAF-AH(II)), however, is a 40-kDa monomer and has an amino acid sequence that exhibits a 41% identity with that of plasma PAF-AH. Although PAF-AH(II) preferentialy hydrolyzes oxidized phospholipids as well as PAF in vitro, the function of this enzyme has not, as yet, been elucidated. Here, we report that PAF-AH(II) functions as an anti-oxidant phospholipase. PAF-AH(II) was found to be an N-myristoylated enzyme that has never been reported among lipases and phospholipases. In MDBK cells treated with oxidants, PAF-AH(II) translocated from cytosol to membranes within 20 min, whereas in cells treated with anti-oxidants, it translocated, conversely, from membranes to cytosol. Overexpression of PAF-AH(II) in Chinese hamster ovary K1 cells suppressed oxidative stress-induced cell death, which occurs by apoptosis. These findings suggest that intracellular PAF-AH(II) translocates between cytosol and membranes in response to a redox state of the cell and protects the cell against oxidative stress most probably by hydrolyzing oxidized phospholipids. PMID- 9405439 TI - Molecular cloning and characterization of human keratan sulfate Gal-6 sulfotransferase. AB - We have previously cloned chondroitin 6-sulfotransferase (C6ST) cDNA from chick embryo chondrocytes. C6ST catalyzes sulfation of chondroitin, keratan sulfate, and sialyl N-acetyllactosamine oligosaccharides. In this study, we report the cloning and characterization of a novel sulfotransferase that catalyzes sulfation of keratan sulfate. This new sulfotransferase cDNA clone was obtained from a human fetal brain library by cross-hybridization with chick C6ST cDNA. The cDNA clone obtained contains a single open reading frame that predicts a type II transmembrane protein composed of 411 amino acid residues. When the cDNA was introduced into a eukaryotic expression vector and transfected in COS-7 cells, keratan sulfate sulfotransferase activity was overexpressed, but C6ST activity was not increased over that of the control. Structural analysis of 35S-labeled glycosaminoglycan, which was formed from keratan sulfate by the reaction with 35S labeled 3'-phosphoadenosine 5'-phosphosulfate and the recombinant sulfotransferase, showed that keratan sulfate was sulfated at position 6 of Gal residues. On the basis of the acceptor substrate specificity, we propose keratan sulfate Gal-6-sulfotransferase (KSGal6ST) for the name of the newly cloned sulfotransferase. KSGal6ST was assigned to chromosome 11p11. 1-11.2 by fluorescence in situ hybridization. Among various human adult tissues, a 2.8 kilobase message of KSGal6ST was expressed mainly in the brain. When poly(A)+ RNAs from the chick embryo cornea and brain were probed with the human KSGal6ST cDNA in Northern hybridization, a clear band with about 2.8 kilobases was detected. These observations suggest that KSGal6ST may participate in the biosynthesis of keratan sulfate in the brain and cornea. PMID- 9405440 TI - The C2 domain of the ubiquitin protein ligase Nedd4 mediates Ca2+-dependent plasma membrane localization. AB - Neuronal precursor cell-expressed developmentally down-regulated 4 (Nedd4) is a ubiquitin protein ligase (E3) containing a hect domain, 3 or 4 WW domains, and a putative C2 domain. We have recently demonstrated an association between the WW domains of Nedd4 and the proline-rich PY motifs (XPPXY) of the epithelial Na+ channel, as well as with PY motifs of several other proteins. The role of the putative C2 domain of Nedd4 has not been elucidated. Here we show that Nedd4, endogenously expressed in Madin-Darby canine kidney cells, was redistributed from the cytosolic to the particulate fraction in response to ionomycin plus Ca2+ treatment. A similar treatment of polarized Madin-Darby canine kidney cells led to an apical and lateral membrane localization of Nedd4, as determined by immunostaining and confocal microscopy. The C2 domain of Nedd4, expressed as a glutathione S-transferase (GST) fusion protein, was sufficient to bind cellular membranes in a Ca2+-dependent manner. Moreover, this GST-Nedd4-C2 domain was able to mediate Ca2+-dependent interactions with phosphatidylserine, phosphatidylinositol, and phosphatidylcholine liposomes in vitro. An epitope tagged Nedd4 lacking its C2 domain and stably expressed in Madin-Darby canine kidney cells failed to mediate the Ca2+-induced plasma membrane localization seen in wild-type (epitope-tagged) Nedd4. These results indicate that the putative C2 domain of Nedd4 acts as a bona fide C2 domain which binds phospholipids and membranes in a Ca2+-dependent fashion and is involved in localizing the protein primarily to the apical region of polarized epithelial cells in response to Ca2+. PMID- 9405441 TI - Purification, cDNA cloning, and gene mapping of the small subunit of human DNA polymerase epsilon. AB - HeLa DNA polymerase epsilon (pol epsilon), possibly involved in both DNA replication and DNA repair, consists of a catalytic subunit of 261 kDa and a tightly bound peptide with a relative molecular mass of 55 kDa. The cDNA of the 261-kDa polypeptide has been independently cloned, sequenced, and then overexpressed in insect cells to give a soluble, but catalytically unstable protein, suggesting that the small subunit of HeLa pol epsilon might be important for stability. HeLa pol epsilon has been isolated by immunoaffinity purification to obtain sequence information which enabled the cloning of a full-length human cDNA encoding the small subunit. The clone encoded nine proteolytic peptides obtained from the subunit. The 59,434-Da predicated polypeptide has 26% identity and 44% homology to the yeast pol epsilon 80-kDa subunit, DPB2. Using fluorescence in situ hybridization, the human pol epsilon p59 locus (DPE2) was assigned to chromosome 14q13-q21. PMID- 9405442 TI - A splice site mutation in the gene of the human type I hair keratin hHa1 results in the expression of a tailless keratin isoform. AB - In this study, we have elucidated the molecular mechanisms underlying the expression of an acidic 41-kDa protein inherited as an autosomal dominant trait of the hair keratin pattern of about 5% of the human population. We show that this protein is a size variant of the large type I hair cortex keratin hHa1 due to a genetic polymorphism in the hHa1 gene. We detected a G-A substitution in the 5' splice site of intron 6 of the hHa1 gene, which segregates with the 41-kDa protein phenotype in two pedigrees and is responsible for the formation of an abnormally spliced hHa1 mRNA species. The use of an alternative 5' splice site leads to the retention of 41 nucleotides of the initial intron 6 sequences in the mature transcript. The open reading frame of the aberrant mRNA creates a premature stop codon immediately downstream of the mutation site. The resulting hHa1 protein variant, hHa1-t, is about 6-kDa smaller than the 47-kDa hHa1 hair keratin and lacks the complete nonhelical tail domain. We show that the tailless hHa1-t is functional, since both recombinant hHa1 and hHa1-t form identical keratin intermediate filaments when assembled in vitro with a type II hair keratin partner. This finding confirms the view of a noninvolvement of the keratin tail domain in filament assembly and explains the lack of a pathological hair phenotype in hHa1-t positive individuals. PMID- 9405443 TI - Nuclear cotransport mechanism of cytoplasmic human MxB protein. AB - Interferon-alpha/beta-inducible Mx proteins belong to the family of large GTPases and share high sequence homology with dynamins in their N-terminal GTP-binding domains. In addition, Mx proteins have a conserved C-terminal leucine zipper element that is involved in their oligomerization. Cytoplasmic human MxA protein mediates resistance to multiple RNA viruses, whereas no antiviral activity has been found for human MxB protein. We have previously shown that MxB protein exists as a nuclear 78-kDa and as a cytoplasmic 76-kDa form in interferon-alpha induced human cells. Using various influenza hemagglutinin epitope-tagged MxB gene constructs in transient transfection experiments in COS-1 cells, we show that the cytoplasmic 76-kDa MxB protein forms hetero-oligomers with the nuclear 78-kDa MxB protein via the C-terminal leucine zipper element. This enables the cytoplasmic form of MxB protein to be translocated into the nucleus together with the nuclear form of MxB protein. This finding was confirmed in interferon-alpha induced HEp-2 and T98G cells transfected with various MxB gene constructs. Cell fractionation studies also suggest that a considerable amount of the cytoplasmic MxB protein is also found in the nucleus. Using confocal laser microscopy, we also demonstrate that the cytoplasmic MxA and the nuclear MxB proteins do not colocalize/oligomerize with each other, and both of these proteins are retained in their specific cellular compartments. PMID- 9405444 TI - Cloning and expression of the rat p8 cDNA, a new gene activated in pancreas during the acute phase of pancreatitis, pancreatic development, and regeneration, and which promotes cellular growth. AB - To characterize at the molecular level the pancreatic emergency program set up by the pancreatic cells in response to pancreatitis, we have developed a strategy in which the phenotype of the pancreatitis affected pancreas is established by characterization of a large number of its transcripts. Herein, we describe the cloning, sequence, and expression of a new gene, named p8, which is strongly activated in pancreatic acinar cells during the acute phase of pancreatitis, in developing pancreas and during pancreatic regeneration. In acinar cells, p8 mRNA is expressed rapidly and specifically in response to cellular pancreatitis induced injury; its induction occurred almost similarly in edematous and necrohemorrhagic pancreatitis, indicating that p8 mRNA is maximally activated even in response to a mild pancreatic injury. Furthermore, in vitro studies suggest that p8 mRNA is induced in pancreatic and non-pancreatic cells in response to some apoptotic stimuli. p8 acts as a promoter of cellular growth factor when its cDNA is transfected into COS-7 and AR4-2J cells. Although we failed to identify p8-related sequences, analysis of its primary and secondary structure suggests that p8 is a basic helix-turn-helix-containing gene with slight homology to several homeotic genes and sufficient signal to be targeted to the nucleus. We therefore propose p8 as a putative transcriptional factor which can regulate pancreatic growth. PMID- 9405445 TI - Irreversible inhibition of lysyl oxidase by homocysteine thiolactone and its selenium and oxygen analogues. Implications for homocystinuria. AB - Homocysteine thiolactone, selenohomocysteine lactone, and homoserine lactone were found to be competitive, irreversible inhibitors of lysyl oxidase, with KI values of 21 +/- 3 microM, 8.3 +/- 2.2 microM, and 420 +/- 56 microM, respectively. The first order rate constants for inactivation (k2) of the enzyme varied over a much smaller range, ranging from 0.12 to 0.18 to 0.28 min-1 for the Se-, thio-, and O lactones, respectively. Mutually exclusive labeling of the enzyme by [1-14C]beta aminopropionitrile, [U-14C]phenylhydrazine, or [35S]homocysteine thiolactone was observed. These labeling results, together with the closely similar perturbations of the near UV-visible spectra of lysyl oxidase and of a model of its lysine tyrosylquinone cofactor by the thiolactone, indicate that the lactones likely derivatize and reduce the active site carbonyl cofactor. Substitution with deuterium at the alpha-carbon of the thiolactone caused a deuterium kinetic isotope effect on k2 of 3.2 +/- 0.2, consistent with the involvement of rate limiting alpha-proton abstraction during lactone-induced inactivation of the enzyme. The activities of plasma amine oxidase and diamine oxidase were only minimally reduced at concentrations of the sulfur or selenium lactones that fully inhibited lysyl oxidase. Thus, these lactones constitute a new category of mechanism-based inactivators selective for lysyl oxidase. Further, these results may relate to the development of connective tissue defects seen in homocystinuria. PMID- 9405447 TI - Molecular cloning, expression, and chromosomal assignment of sarcolemmal associated proteins. A family of acidic amphipathic alpha-helical proteins associated with the membrane. AB - Two overlapping cDNAs encoding a novel sarcolemmal associated protein (SLAP) were isolated from a cardiac cDNA expression library by immunoscreening with anti sarcolemmal antibodies. Further characterization of these clones showed that they belonged to a family of related cDNAs that potentially encode polypeptides of 37, 46, and 74 kDa designated SLAP1, SLAP2, and SLAP3, respectively. The SLAP3 transcript was ubiquitously expressed, whereas SLAP1 and SLAP2 transcripts were predominantly expressed in cardiac, soleus, and smooth muscle. SLAP was encoded by a single gene that mapped to chromosome 3p14.3-21.2, and the various transcripts are likely generated by alternative splicing. The primary structure of SLAP predicted that it would have large regions of coiled-coil structure including an 11-heptad acidic amphipathic alpha-helical segment. The carboxyl terminal region of the SLAP proteins was predicted to have a transmembrane domain, although there was no discernible signal sequence. SLAPs could only be solubilized from cardiac membrane with detergents suggesting that they were integral membrane proteins. Subcellular distribution studies showed that MYC epitope-tagged SLAP localized to regions of juxtaposition between neighboring cell membranes although an intracellular pool of the protein was also present in cells undergoing apparent cleavage. Immunohistochemical localization of SLAP in cardiac muscle revealed that SLAP associated with the sarcolemma and also displayed a reticular pattern of staining that resembled the transverse tubules and the sarcoplasmic reticulum. The SLAPs define a new family of tail-anchored membrane proteins that exhibit tissue-specific expression and are uniquely situated to serve a variety of roles through their coiled-coil motifs. PMID- 9405446 TI - Activation of stress-activated protein kinases/c-Jun N-terminal protein kinases (SAPKs/JNKs) by a novel mitogen-activated protein kinase kinase. AB - Mitogen-activated protein kinase (MAPK) kinases (MKKs) are dual-specificity protein kinases that phosphorylate and activate MAPK. We have isolated a cDNA encoding a novel protein kinase that has significant homology to MKKs. The novel kinase MKK7 has a nucleotide sequence that encodes an open reading frame of 347 amino acids with 11 kinase subdomains. MKK7 is ubiquitously expressed in all adult and embryonic organs but displays high expression in epithelial tissues at later stages of fetal development. When transiently expressed in 293 cells, MKK7 specifically activated stress-activated protein kinases (SAPKs)/c-Jun N-terminal protein kinases (JNKs) but not extracellular-regulated kinase or p38 kinase. A kinase-negative mutant of MKK7 inhibits interleukin-1beta, lipopolysaccharide, and MEKK1-induced SAPK/JNK activation. Thus, MKK7 is a new member of the MAPK kinase family that functions upstream of SAPK/JNK in the SAPK/JNK signaling pathway. PMID- 9405448 TI - Regulated human erythropoietin receptor expression in mouse brain. AB - Erythropoietin (Epo) is known for its role in erythropoiesis and acts by binding to its receptor (EpoR) on the surface of erythroid progenitors. EpoR activity follows the site of hematopoiesis from the embryonic yolk sac to the fetal liver and then the adult spleen and bone marrow. Expression of EpoR has also been observed in selected cells of non-hematopoietic origin, such as the embryonic mouse brain during mid-gestation, at levels comparable to adult bone marrow. EpoR transcripts in brain decrease during development falling by birth to less than 1 3% of the level in hematopoietic tissue. We have now recapitulated this pattern of expression using a human EpoR transgene consisting of an 80-kb human EpoR genomic fragment. The highest level of expression was observed in the embryonic yolk sac and fetal liver, analogous to the endogenous gene, in addition to expression in adult spleen and bone marrow. Although activity of this transgene in brain is initially lower than the endogenous gene, it does exhibit the down regulation observed for the endogenous gene in adult brain. The expression pattern of hybrid transgenes of an hEpoR promoter fused to beta-galactosidase in 9. 5-day embryos suggested that the hEpoR promoter region between -1778 and -150 bp 5' of the transcription start site is necessary to direct EpoR expression in the neural tube. EpoR expression in the neural tube may be the origin of the EpoR transcripts detected in brain during development. These data demonstrate that both the mouse and human EpoR genes contain regulatory elements to direct significant levels of expression in a developmentally controlled manner in brain and suggest that in addition to its function during erythropoiesis, EpoR may play a role in the development of selected non-hematopoietic tissue. PMID- 9405449 TI - TWEAK, a new secreted ligand in the tumor necrosis factor family that weakly induces apoptosis. AB - The members of the tumor necrosis factor (TNF) family play pivotal roles in the regulation of the immune system. Here we describe a new ligand in this family, designated TWEAK. The mouse and human versions of this protein are unusually conserved with 93% amino acid identity in the receptor binding domain. The protein was efficiently secreted from cells indicating that, like TNF, TWEAK may have the long range effects of a secreted cytokine. TWEAK transcripts were abundant and found in many tissues, suggesting that TWEAK and TRAIL belong to a new group of widely expressed ligands. Like many members of the TNF family, TWEAK was able to induce interleukin-8 synthesis in a number of cell lines. The human adenocarcinoma cell line, HT29, underwent apoptosis in the presence of both TWEAK and interferon-gamma. Thus, TWEAK resembles many other TNF ligands in the capacity to induce cell death; however, the fact that TWEAK-sensitive cells are relatively rare suggests that TWEAK along with lymphotoxins alpha/beta and possibly CD30L trigger death via a weaker, nondeath domain-dependent mechanism. PMID- 9405450 TI - HgCl2-induced interleukin-4 gene expression in T cells involves a protein kinase C-dependent calcium influx through L-type calcium channels. AB - Mercuric chloride (HgCl2) induces T helper 2 (Th2) autoreactive anti-class II T cells in Brown Norway rats. These cells produce interleukin (IL)-4 and induce a B cell polyclonal activation that is responsible for autoimmune disease. In Brown Norway rats, HgCl2 triggers early IL-4 mRNA expression both in vivo and in vitro by T cells, which may explain why autoreactive anti-class II T cells acquire a Th2 phenotype. The aim of this study was to explore the transduction pathways by which this chemical operates. By using two murine T cell hybridomas that express IL-4 mRNA upon stimulation with HgCl2, we demonstrate that: 1) HgCl2 acts at the transcriptional level without requiring de novo protein synthesis; 2) HgCl2 induces a protein kinase C-dependent Ca2+ influx through L-type calcium channels; 3) calcium/calcineurin-dependent pathway and protein kinase C activation are both implicated in HgCl2-induced IL-4 gene expression; and 4) HgCl2 can activate directly protein kinase C, which might be one of the main intracellular target for HgCl2. These data are in agreement with an effect of HgCl2 which is independent of antigen-specific recognition. It may explain the T cell polyclonal activation in the mercury model and the expansion of pathogenic autoreactive anti class II Th2 cells in this context. PMID- 9405451 TI - Identification of chondromodulin I as a novel endothelial cell growth inhibitor. Purification and its localization in the avascular zone of epiphyseal cartilage. AB - Cartilage is unique among tissues of mesenchymal origin in that it is resistant to vascular invasion due to an intrinsic angiogenic inhibitor. During endochondral bone formation, however, calcified cartilage formed in the center of the cartilaginous bone rudiment allows vascular invasion, which initiates the replacement of cartilage by bone. The transition of cartilage from the angioresistant to the angiogenic status thus plays a key role in bone formation. However, the molecular basis of this phenotypic transition of cartilage has been obscure. We report here purification of an endothelial cell growth inhibitor from a guanidine extract of bovine epiphyseal cartilage. The N-terminal amino acid sequence indicated that the inhibitor was identical to chondromodulin I (ChM-I), a cartilage-specific growth-modulating factor. Purified ChM-I inhibited DNA synthesis and proliferation of vascular endothelial cells as well as tube morphogenesis in vitro. Expression of ChM-I cDNA in COS7 cells indicated that mature ChM-I molecules were secreted from the cells after post-translational modifications and cleavage from the transmembrane precursor at the predicted processing signal. Recombinant ChM-I stimulated DNA synthesis and proteoglycan synthesis of cultured growth plate chondrocytes, but inhibited tube morphogenesis of endothelial cells. In situ hybridization and immunohistochemical studies indicated that ChM-I is specifically expressed in the avascular zone of cartilage in developing bone, but not present in calcifying cartilage. These results suggest a regulatory role of ChM-I in vascular invasion during endochondral bone formation. PMID- 9405452 TI - Molecular genetic identification of a pathway for heme binding to cytochrome b6. AB - Heme binding to cytochrome b6 is resistant, in part, to denaturing conditions that typically destroy the noncovalent interactions between the b hemes and their apoproteins, suggesting that one of two b hemes of holocytochrome b6 is tightly bound to the polypeptide. We exploited this property to define a pathway for the conversion of apo- to holocytochrome b6, and to identify mutants that are blocked at one step of this pathway. Chlamydomonas reinhardtii strains carrying substitutions in either one of the four histidines that coordinate the bh or bl hemes to the apoprotein were created. These mutations resulted in the appearance of distinct immunoreactive species of cytochrome b6, which allowed us to specifically identify cytochrome b6 with altered bh or bl ligation. In gabaculine treated (i.e. heme-depleted) wild type and site-directed mutant strains, we established that (i) the single immunoreactive band, observed in strains carrying the bl site-directed mutations, corresponds to apocytochrome b6 and (ii) the additional band present in strains carrying bh site-directed mutations corresponds to a bl-heme-dependent intermediate in the formation of holocytochrome b6. Five nuclear mutants (ccb strains) that are defective in holocytochrome b6 formation display a phenotype that is indistinguishable from that of strains carrying site-directed bh ligand mutants. The defect is specific for cytochrome b6 assembly, because the ccb strains can synthesize other b cytochromes and all c-type cytochromes. The ccb strains, which define four nuclear loci (CCB1, CCB2, CCB3, and CCB4), provide the first evidence that a b type cytochrome requires trans-acting factors for its heme association. PMID- 9405453 TI - A primary role for K+ and Na+ efflux in the activation of apoptosis. AB - Cell shrinkage is a major characteristic of apoptosis, but the mechanism and role of this process in cell death are poorly understood. The primary factor that controls volume regulation in all cells is ions, and thus we have examined the movement of ions at the single cell level in lymphocytes during apoptosis. Activation of the death program with several stimuli that act through independent pathways to stimulate apoptosis results in a synchronous shift of cells from a normal cell size to a shrunken cell size. Only the shrunken cells exhibit DNA fragmentation and an approximate 4-fold elevation of caspase-3-like activity. Analysis of K+ and Na+ ion content of individual cells by flow cytometry revealed that the intracellular ionic strength of apoptotic cells decreased substantially from their non-shrunken counterparts. Additionally, we show apoptosis is enhanced under conditions where the intracellular K+ concentration is diminished and that apoptosis is inhibited when K+ efflux is prevented. These data show that the efflux of ions, primarily potassium, plays a necessary and perhaps a pivotal role in the cell death program. PMID- 9405454 TI - Activation of protein-tyrosine kinase Pyk2 is downstream of Syk in FcepsilonRI signaling. AB - Aggregation of the FcepsilonRI, a member of the immune receptor family, induces the activation of proteintyrosine kinases and results in tyrosine phosphorylation of proteins that are involved in downstream signaling pathways. Here we report that Pyk2, another member of the focal adhesion kinase family, was present in the RBL-2H3 mast cell line and was rapidly tyrosine-phosphorylated and activated after FcepsilonRI aggregation. Tyrosine phosphorylation of Pyk2 was also induced by the calcium ionophore A23187, by phorbol myristate acetate, or by stimulation of G-protein-coupled receptors. Adherence of cells to fibronectin dramatically enhanced the induced tyrosine phosphorylation of Pyk2. Although Src family kinases are activated by FcepsilonRI stimulation and tyrosine-phosphorylate the receptor subunits, the activation and tyrosine phosphorylation of Pyk2 were downstream of Syk. In contrast, tyrosine phosphorylation of Pyk2 by stimulation of G-protein-coupled receptors was independent of Syk. Therefore, the FcepsilonRI induced tyrosine phosphorylation of Pyk2 is downstream of Syk and may play a role in cell secretion. PMID- 9405455 TI - Vinculin is associated with the E-cadherin adhesion complex. AB - Cadherins mediate calcium-dependent cell-cell adhesion, and this activity is regulated by cytoplasmic interactions between cadherins, catenins, and the actin based cytoskeleton. alpha-Catenin plays a critical role in the transmembrane anchorage of cadherins, and deletion of alpha-catenin has been shown to inactivate cadherin-mediated adhesion, resulting in a nonadhesive phenotype. Here we show that serum starvation increases E-cadherin expression and induces E cadherin-dependent adhesion in the MDA-MB-468 breast cancer cell line. This adhesion occurred despite a lack of alpha-catenin expression, which was caused by mutations in the alpha-catenin gene. Coprecipitation analysis suggests that this adhesion may be mediated by cytoplasmic connections from cadherins to the cytoskeleton involving vinculin. A high level of vinculin associated with E cadherin immunoprecipitates was observed in MDA-MB-468 cells. In contrast, vinculin was not detected in E-cadherin complexes in the A431 and MCF-7 epithelial carcinoma cell lines, which express alpha-catenin. However, in reciprocal immunoprecipitations using anti-vinculin antibodies, E-cadherin associated strongly with vinculin in MDA-MB-468 cells and, to a lesser extent, in A431 and MCF-7 cells. These results suggest that both alpha-catenin and vinculin may be present in the adhesion complex. To test the hypothesis that vinculin associates with E-cadherin complexes via beta-catenin, excess recombinant beta catenin or alpha-catenin fusion protein was added to MDA-MB-468 cell lysates. Both specifically inhibited the coprecipitation of E-cadherin with vinculin, suggesting competition for the same binding site. These results suggest that vinculin plays a role in the establishment or regulation of the cadherin-based cell adhesion complex by direct interaction with beta-catenin. PMID- 9405456 TI - Finkel-Biskis-Reilly osteosarcoma virus v-Fos inhibits adipogenesis and both the activity and expression of CCAAT/enhancer binding protein alpha, a key regulator of adipocyte differentiation. AB - Finkel-Biskis-Reilly (FBR) osteosarcoma virus v-Fos causes tumors of mesenchymal origin, including osteosarcomas, rhabdomyosarcomas, chondrosarcomas, and liposarcomas. Because the cell of origin in all these tumors is a pluripotent mesenchymal cell, the variety of tumors seen in mice which express FBR v-Fos implies that FBR v-Fos inhibits multiple differentiation pathways. To study the mechanism of FBR v-Fos' inhibition of mesenchymal differentiation, we utilized an in vitro model of adipocyte differentiation. We show by both morphological and biochemical means that FBR v-Fos inhibits adipocyte differentiation in vitro. This inhibition is due to FBR v-Fos' inhibition of the growth arrest characteristic of terminal differentiation and FBR v-Fos' inhibition of the expression and activity of a key regulator of this growth arrest, C/EBPalpha. The in vitro inhibition of adipogenesis by FBR v-Fos has in vivo significance as immunostaining of FBR v-Fos-induced tumors shows no CCAAT/enhancer binding protein (EBP)-alpha expression. These data implicate C/EBPalpha as a protein involved in the generation of liposarcomas. PMID- 9405457 TI - Locations of calmodulin and FK506-binding protein on the three-dimensional architecture of the skeletal muscle ryanodine receptor. AB - Isolated skeletal muscle ryanodine receptors (RyRs) complexed with the modulatory ligands, calmodulin (CaM) or 12-kDa FK506-binding protein (FKBP12), have been characterized by electron cryomicroscopy and three-dimensional reconstruction. RyRs are composed of 4 large subunits (molecular mass 565 kDa) that assemble to form a 4-fold symmetric complex that, architecturally, comprises two major substructures, a large ( approximately 80% of the total mass) cytoplasmic assembly and a smaller transmembrane assembly. Both CaM and FKBP12 bind to the cytoplasmic assembly at sites that are 10 and 12 nm, respectively, from the putative entrance to the transmembrane ion channel. FKBP12 binds along the edge of the square-shaped cytoplasmic assembly near the face that interacts in vivo with the sarcolemma/transverse tubule membrane system, whereas CaM binds within a cleft that faces the junctional face of the sarcoplasmic reticulum membrane at the triad junction. Both ligands interact with a domain that connects directly to a cytoplasmic extension of the transmembrane assembly of the receptor, and thus might cause structural changes in the domain which in turn modulate channel gating. PMID- 9405458 TI - Molecular cloning and characterization of lustrin A, a matrix protein from shell and pearl nacre of Haliotis rufescens. AB - A specialized extracellular matrix of proteins and polysaccharides controls the morphology and packing of calcium carbonate crystals and becomes occluded within the mineralized composite during formation of the molluscan shell and pearl. We have cloned and characterized the cDNA coding for Lustrin A, a newly described matrix protein from the nacreous layer of the shell and pearl produced by the abalone, Haliotis rufescens, a marine gastropod mollusc. The full-length cDNA is 4,439 base pairs (bp) long and contains an open reading frame coding for 1,428 amino acids. The deduced amino acid sequence reveals a highly modular structure with a high proportion of Ser (16%), Pro (14%), Gly (13%), and Cys (9%). The protein contains ten highly conserved cysteine-rich domains interspersed by eight proline-rich domains; a glycine- and serine-rich domain lies between the two cysteine-rich domains nearest the C terminus, and these are followed by a basic domain and a C-terminal domain that is highly similar to known protease inhibitors. The glycine- and serine-rich domain and at least one of the proline rich domains show sequence similarity to proteins of two extracellular matrix superfamilies (one of which also is involved in the mineralized matrixes of bone, dentin, and avian eggshell). The arrangement of alternating cysteine-rich domains and proline-rich domains is strikingly similar to that found in frustulins, the proteins that are integral to the silicified cell wall of diatoms. Its modular structure suggests that Lustrin A is a multifunctional protein, whereas the occurrence of related sequences suggest it is a member of a multiprotein family. PMID- 9405459 TI - G protein-coupled receptor kinase 5 in cultured vascular smooth muscle cells and rat aorta. Regulation by angiotensin II and hypertension. AB - GRK5, a recently cloned member of the G protein-coupled receptor kinase family, has been shown to phosphorylate and participate in the desensitization of angiotensin II (Ang II) type 1A (AT1A) receptors. In this study, the effect of angiotensin II on GRK5 expression was examined in cultured vascular smooth muscle cells and aortas of Ang II-infused hypertensive rats. In vascular smooth muscle cells, Ang II (100 nM) up-regulated GRK5 mRNA as early as 1 h, with a peak at 16 h. This up-regulation was dose- and calcium-dependent. The increase in GRK5 mRNA was reflected in a smaller increase in protein expression, which nonetheless had functional significance since AT1 receptor phosphorylation was increased and phospholipase C activation was decreased following prolonged incubation with Ang II. In aortas of Ang II-infused hypertensive rats, both GRK5 mRNA and protein levels increased approximately 3-fold compared with sham-operated rats at 5 and 7 days, respectively. This up-regulation was blocked either by losartan or by the nonspecific vasodilator hydralazine. Since a subpressor dose of Ang II did not increase GRK5 mRNA levels and norepinephrine infusion also increased GRK5 mRNA expression, we conclude that Ang II-induced GRK5 up-regulation in rat aortas may be due to hypertension per se. Hormone- and hemodynamic stress-induced GRK5 regulation may provide a novel molecular basis for long-term regulation of agonist sensitivity of vascular cells. PMID- 9405460 TI - Molecular characteristics of the novel intermediate filament protein paranemin. Sequence reveals EAP-300 and IFAPa-400 are highly homologous to paranemin. AB - Paranemin was initially found to copurify with the intermediate filament (IF) proteins vimentin and desmin from embryonic chick skeletal muscle and was described as an IF-associated protein (IFAP). We have purified paranemin from embryonic chick skeletal muscle, prepared antibodies, and demonstrated that they label at the Z-lines of both adult avian and porcine cardiac and skeletal muscle myofibrils. We determined the cDNA sequence of paranemin by immunoscreening a lambdagt22A cDNA library from embryonic chick skeletal muscle. Northern blot analysis revealed a single transcript of 5.3 kilobases, which is much smaller than predicted from the size of paranemin (280 kDa) by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The derived amino acid sequence of paranemin (1,606 residues; 178,161 kDa) contains the conserved IF rod domain (308 amino acids), which has highest homology to the rod domains of nestin and tanabin. Thus, paranemin is an IF protein rather than an IFAP. Sequence analysis also revealed that the partial cDNA sequences of two proteins, namely EAP-300 and IFAPa-400, are almost identical to regions of the cDNA sequence of paranemin. The complete paranemin cDNA was expressed in a cell line (SW13) with, and without, detectable cytoplasmic IFs. Antibody labeling of these cells suggests that paranemin does not form IFs by itself, but rather is incorporated into heteropolymeric IFs with vimentin. PMID- 9405461 TI - Identification of a novel cis-element required for the constitutive activity and osmotic response of the rat aldose reductase promoter. AB - A new and essential cis-element AEE (aldose reductase enhancer element), necessary for the constitutive activity and the osmotic stress response of rat aldose reductase transcription in a rat liver cell line, has been identified. In transient transfection assays, an increase in promoter activity, up to 3.8-fold, was observed with osmotic stress (600 mosm/kg H2O) using a luciferase reporter gene construct containing aldose reductase promoter sequence from -1,094 base pair (bp) to +23 bp. A deletion between -1,071 and -895 bp reduced the constitutive activity and abolished the osmotic response of the promoter. Exonuclease III mediated in vivo DNA footprinting and dimethyl sulfate in vivo footprinting revealed DNA protection of a 32-bp region and two guanosines (G) within this region protected from methylation, respectively. Electrophoretic gel mobility shift assays using whole liver cell extracts showed protein binding, under both normal and stressed conditions. Deletion of the sequence between the two guanosines protected by in vivo dimethyl sulfate DNA footprinting (GAAGAGTG) in a luciferase construct (-1,094 bp to +23 bp) abolished the constitutive promoter activity. One copy of AEE fused to the thymidine kinase promoter gave a maximum constitutive activity of 7.7-fold and a maximum osmotic response activity of 6. 7-fold. PMID- 9405462 TI - Modulation of the arrestin-clathrin interaction in cells. Characterization of beta-arrestin dominant-negative mutants. AB - We recently demonstrated that nonvisual arrestins interact via a C-terminal binding domain with clathrin and function as adaptor proteins to promote beta2 adrenergic receptor (beta2AR) internalization. Here, we investigated the potential utility of a mini-gene expressing the clathrin-binding domain of beta arrestin (beta-arrestin (319-418)) to function as a dominant-negative with respect to beta2AR internalization and compared its properties with those of beta arrestin and beta-arrestin-V53D, a previously reported dominant-negative mutant. In vitro studies demonstrated that beta-arrestin-V53D bound better to clathrin than beta-arrestin but was significantly impaired in its interaction with phosphorylated G protein-coupled receptors. In contrast, whereas beta-arrestin (319-418) also bound well to clathrin it completely lacked receptor binding activity. When coexpressed with the beta2AR in HEK293 cells, beta-arrestin (319 418) effectively inhibited agonist-promoted receptor internalization, whereas beta-arrestin-V53D was only modestly effective. However, both constructs significantly inhibited the stimulation of beta2AR internalization by beta arrestin in COS-1 cells. Interestingly, immunofluorescence microscopy analysis reveals that both beta-arrestin (319-418) and beta-arrestin-V53D are constitutively localized in clathrin-coated pits in COS-1 cells. These results indicate the potential usefulness of beta-arrestin (319-418) to effectively block arrestin-clathrin interaction in cells and suggest that this construct may prove useful in further defining the mechanisms involved in G protein-coupled receptor trafficking. PMID- 9405463 TI - The highly conserved Stt3 protein is a subunit of the yeast oligosaccharyltransferase and forms a subcomplex with Ost3p and Ost4p. AB - The oligosaccharyltransferase has been purified from Saccharomyces cerevisiae as an hetero-oligomeric complex composed of four or six subunits. Here, the in vivo subunit composition and stoichiometry of the oligosaccharyltransferase were investigated by attaching an epitope coding sequence to a previously characterized subunit gene, OST3. Five (Ost1p, Wbp1p, Swp1p, Ost2p, and Ost5p) of the seven polypeptides that were coimmunoprecipitated with the epitope-tagged Ost3p were identical to those obtained by the conventional purification procedure. Two additional coprecipitating polypeptides with apparent molecular masses of 60 and 3.6 kDa were identified as the 78-kDa Stt3 protein and the 36 residue Ost4 protein, respectively. Stt3p and Ost4p were previously identified in screens for gene products involved in N-linked glycosylation. Quantification of the in vivo radiolabeled subunits and the radioiodinated purified enzyme shows that the yeast oligosaccharyltransferase is composed of equimolar amounts of eight subunits. Exposure of the immunoprecipitated oligosaccharyltransferase to mild protein denaturants yielded a subcomplex comprised of Stt3p, Ost3p, and Ost4p. These experiments, taken together with genetic and biochemical evidence for subunit interactions, suggest that the enzyme is composed of the following three subcomplexes: (a) Stt3p-Ost4p-Ost3p, (b) Swp1p-Wbp1p-Ost2p, and (c) Ost1p Ost5p. PMID- 9405464 TI - The vascular endothelial growth factor receptor KDR activates multiple signal transduction pathways in porcine aortic endothelial cells. AB - Vascular endothelial growth factor A (here referred to as VEGF) is an endothelium specific growth factor that binds to two distinct receptor tyrosine kinases, designated Flt-1 and KDR/Flk-1. VEGF stimulates autophosphorylation of both receptors, but little is known about their signal transduction properties. In this study, we used porcine aortic endothelial (PAE) cells overexpressing KDR (PAE/KDR) to evaluate the interaction of KDR with intracellular proteins and compared them with Flt-1-expressing PAE cells (PAE/Flt-1). VEGF-induced stimulation of KDR results in the association and phosphorylation of the 46-, 52 , and 66-kDa isoforms of Shc and the induction of Shc-Grb2 complex formation. In a similar fashion, KDR associates with Grb2 and Nck in a ligand-dependent fashion, suggesting Shc, Grb2, and Nck as potential candidates involved in the regulation of endothelial function. Another strong candidate is mitogen-activated protein (MAP) kinase, which is strongly activated in response to VEGF stimulation as demonstrated by phosphorylation of the specific substrate myelin basic protein. Inhibition of MAP kinase activation by PD98059, a specific MAP kinase kinase inhibitor, results in inhibition of VEGF-induced proliferation of PAE/KDR cells. In contrast, VEGF-induced stimulation of Flt-1 does not activate MAP kinase in PAE/Flt-1 cells. In this study we provide the first two examples of molecules potentially capable of functionally counteracting the endothelial response to VEGF, namely SHP-1 and SHP-2. These two SH2 protein-tyrosine phosphatases physically associate with KDR secondary to VEGF stimulation, raising the interesting possibility that both molecules participate in the generation and/or modulation of VEGF-induced signals. Taken together, our results substantially broaden the spectrum of KDR-associating molecules, indicating that endothelial function and angiogenesis are regulated by a diverse network of signal transduction cascades. PMID- 9405465 TI - Phosphorylation of the alpha-amino-3-hydroxy-5-methylisoxazole4-propionic acid receptor GluR1 subunit by calcium/calmodulin-dependent kinase II. AB - Modulation of alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic Acid (AMPA) receptors in the brain by protein phosphorylation may play a crucial role in the regulation of synaptic plasticity. Previous studies have demonstrated that calmodulin (CaM) kinase II can phosphorylate and modulate AMPA receptors. However, the sites of CaM kinase phosphorylation have not been unequivocally identified. In the current study, we have generated two phosphorylation site specific antibodies to analyze the phosphorylation of the glutamate receptor GluR1 subunit. These antibodies recognize GluR1 only when it is phosphorylated on serine residues 831 or 845. We have used these antibodies to demonstrate that serine 831 is specifically phosphorylated by CaM kinase II in transfected cells expressing GluR1 as well as in hippocampal slice preparations. Two-dimensional phosphopeptide mapping experiments indicate that Ser-831 is the major site of CaM kinase II phosphorylation on GluR1. In addition, treatment of hippocampal slice preparations with phorbol esters and forskolin increase the phosphorylation of serine 831 and 845, respectively, indicating that protein kinase C and protein kinase A phosphorylate these residues in hippocampal slices. These results identify the site of CaM kinase phosphorylation of the GluR1 subunit and demonstrate that GluR1 is multiply phosphorylated by protein kinase A, protein kinase C, and CaM kinase II in situ. PMID- 9405466 TI - epsilon-Sarcoglycan, a broadly expressed homologue of the gene mutated in limb girdle muscular dystrophy 2D. AB - The sarcoglycans are transmembrane components of the dystrophin-glycoprotein complex, which links the cytoskeleton to the extracellular matrix in adult muscle fibers. Mutations in all four known sarcoglycan genes (alpha, beta, gamma, and delta) have been found in humans with limb-girdle muscular dystrophy. We have identified a novel protein, epsilon-sarcoglycan, that shares 44% amino acid identity with alpha-sarcoglycan (adhalin). We show that epsilon-sarcoglycan is a membrane-associated glycoprotein and document its expression by Northern blotting, immunoblotting, and immunofluorescence. In contrast to alpha-delta sarcoglycans, which are expressed predominantly or exclusively in striated muscle, epsilon-sarcoglycan is broadly distributed in muscle and nonmuscle cells of both embryos and adults. These results raise the possibility that sarcoglycan containing complexes mediate membrane-matrix interactions in many cell types. PMID- 9405467 TI - Molecular cloning, expression, and characterization of the authentic hyaluronan synthase from group C Streptococcus equisimilis. AB - We previously reported the first cloning of a functional glycosaminoglycan synthase, the hyaluronan synthase (HAS) from Group A Streptococcus pyogenes (spHAS) (DeAngelis, P. L., Papaconstantinou, J., and Weigel, P. H. (1993) J. Biol. Chem. 268, 19181-19184). Group A spHAS was unrelated to a putative Group C HA synthase reported by others (Lansing, M., Lellig, S., Mausolf, A., Martini, I. , Crescenzi, F., Oregon, M., and Prehm, P. (1993) Biochem. J. 289, 179-184). Here we report the isolation of a bona fide HA synthase gene from a highly encapsulated strain of Group C Streptococcus equisimilis. The encoded protein, designated seHAS, is 417 amino acids long (calculated molecular weight, 47,778; calculated pI, 9.1) and is the smallest member of the HAS family identified thus far. The enzyme migrates anomalously fast in SDS-polyacrylamide gel electrophoresis (approximately 42,000 Da). The seHAS protein shows no similarity (<2% identity) to the previously reported Group C gene, which is not an HA synthase. The seHAS and spHAS protein and coding sequences are 72 and 70% identical, respectively. seHAS is also similar to eukaryotic HAS1 (approximately 31% identical), HAS2 (approximately 28% identical), and HAS3 (28% identical). The deduced protein sequence of seHAS was confirmed by reactivity with a synthetic peptide antibody. Recombinant seHAS expressed in Escherichia coli was recovered in membranes as a major protein (approximately 10% of the total protein) and synthesized very large HA (Mr >7 x 10(6)) in the presence of UDP-GlcNAc and UDP GlcA. The product contained equimolar amounts of both sugars and was degraded by the specific Streptomyces hyaluronidase. Comparison of the two recombinant streptococcal enzymes in isolated membranes showed that seHAS and spHAS are essentially identical in the steady-state size distribution of HA chains they synthesize, but seHAS has an intrinsic 2-fold faster rate of chain elongation (Vmax) than spHAS. seHAS is the most active HA synthase identified thus far; it polymerizes HA at an average rate of 160 monosaccharides/s. The two bacterial HA synthase genes may have arisen from a common ancient gene shared with the early evolving vertebrates. PMID- 9405468 TI - The mammalian target of rapamycin phosphorylates sites having a (Ser/Thr)-Pro motif and is activated by antibodies to a region near its COOH terminus. AB - The eukaryotic initiation factor 4E (eIF4E)-binding protein, PHAS-I, was phosphorylated rapidly and stoichiometrically when incubated with [gamma-32P]ATP and the mammalian target of rapamycin (mTOR) that had been immunoprecipitated with an antibody, mTAb1, directed against a region near the COOH terminus of mTOR. PHAS-I was phosphorylated more slowly by mTOR obtained either by immunoprecipitation with other antibodies or by affinity purification using a rapamycin/FKBP12 resin. Adding mTAb1 to either of these preparations of mTOR increased PHAS-I phosphorylation severalfold, indicating that mTAb1 activates the mTOR protein kinase. mTAb1-activated mTOR phosphorylated Thr36, Thr45, Ser64, Thr69, and Ser82 in PHAS-I. All five of these sites fit a (Ser/Thr)-Pro motif and are dephosphorylated in response to rapamycin in rat adipocytes. Thus, our findings indicate that Pro is a determinant of the mTOR protein kinase specificity and that mTOR contributes to the phosphorylation of PHAS-I in cells. PMID- 9405469 TI - Differential splicing and alternative polyadenylation generate multiple mimecan mRNA transcripts. AB - We previously showed the 25-kDa corneal keratan sulfate proteoglycan to be a translation product of the gene producing osteoglycin and proposed the name mimecan for this gene and its product. We also demonstrated three mimecan RNA transcripts using Northern blot analysis. In this report, we investigate the mechanisms accounting for these transcripts. Ribonuclease protection analysis and reverse transcription-polymerase chain reaction of bovine corneal mRNA detected a mimecan transcript that lacked 278 base pairs of the 5'-untranslated region between residues 62 and 340. This splice variant represents the predominant form of mimecan mRNA in bovine cornea and sclera. It was also detectable in other bovine tissues as a minor transcript. Two additional cDNA clones that were isolated contained 398 bases of nucleotide sequence at the 3'-end of mimecan cDNA, not present in the published sequence. Ribonuclease protection analyses with the 3'-probe, which included the new sequence, allow detection of three RNA transcripts while 5'-probes recognized only two. These results indicate that the three canonical polyadenylation sites in the 3'-untranslated region of mimican mRNA are alternatively selected. Possible roles for this previously undetected degree of diversity of mimecan RNA isoforms transcribed in the same tissue are discussed. PMID- 9405470 TI - A proline residue in the alpha-helical rod domain of type I keratin 16 destabilizes keratin heterotetramers. AB - The type I keratins 14 (K14) and 16 (K16) are distinct in their assembly properties and their expression pattern despite a high degree of sequence identity. Understanding K16 function and regulation is of interest, given its strong induction in keratinocytes located at the wound edge after injury to stratified epithelia. We reported previously that, compared with K14, K16 forms unstable heterotetramers with either K5 or K6 as the type II keratin pairing partner (Paladini, R. D., Takahashi, K., Bravo, N. S., and Coulombe, P. A. (1996) J. Cell Biol. 132, 381-397). We show here that yet another related type I keratin, K17, forms stable heterotetramers with a variety of type II keratins, further accentuating the unique nature of K16. Analysis of chimeric K14-K16 proteins in a heterotetramer formation assay indicated that the instability determinant resides in a 220-amino acid segment within the alpha-helical rod domain of K16. Site-directed mutagenesis revealed that Pro188, an amino acid residue located in subdomain 1B of the rod, accounts quantitatively for the instability of K16-containing heterotetramers under denaturing conditions. In vitro polymerization studies suggest that the presence of Pro188 correlates with a reduction in assembly efficiency. In addition to their implications for the stable conformation of the keratin heterotetramers, these findings suggest that the tetramer-forming properties of K16 may influence its partitioning between the soluble and polymer pools, and hence contribute to its regulation in epithelial cells under resting and wound repair conditions. PMID- 9405471 TI - Involvement of yeast sphingolipids in the heat stress response of Saccharomyces cerevisiae. AB - A role for sphingolipids in the yeast heat stress response has been suggested by the isolation of suppressors of mutants lacking these lipids, which are unable to grow at elevated temperatures. The current study examines the possible role of sphingolipids in the heat adaptation of yeast cells as monitored by growth and viability studies. The suppressor of long chain base auxotrophy (SLC, strain 7R4) showed a heat-sensitive phenotype that was corrected by transformation with serine palmitoyltransferase. Thus, the deficiency in sphingolipids and not the suppressor mutation was the cause of the heat-sensitive phenotype of the SLC strain 7R4. The ability of sphingolipids to rescue the heat-sensitive phenotype was examined, and two endogenous yeast sphingoid backbones, phytosphingosine and dihydrosphingosine, were found to be most potent in this effect. Next, the effect of heat stress on the levels of the three major classes of sphingolipids was determined. The inositol phosphoceramides showed no change over a 1.5-h time course. However, the four detected species of sphingoid bases increased after 15 min of heat stress from 1.4- to 10.8-fold. The largest increases were seen in two sphingoid bases, C20 phytosphingosine and C20 dihydrosphingosine, which increased 6.4- and 10.8-fold over baseline, respectively. At 60 min of heat stress two species of yeast ceramide increased by 9.2- and 10.6-fold over baseline. The increase seen in the ceramides was partially decreased by Fumonisin B1, a ceramide synthase inhibitor. Therefore, heat stress induces accumulation of sphingoid bases and of ceramides, probably through de novo synthesis. Taken together, these results demonstrate that sphingolipids are involved in the yeast heat stress adaptation. PMID- 9405472 TI - The p43 component of the mammalian multi-synthetase complex is likely to be the precursor of the endothelial monocyte-activating polypeptide II cytokine. AB - p43 is one of the three auxiliary components invariably associated with nine aminoacyl-tRNA synthetases as a multienzyme complex ubiquitous to all eukaryotic cells from flies to humans. The cDNA encoding the hamster protein was isolated by using mixed oligonucleotides deduced from peptide sequences. The 359-amino acid protein is the hamster homologue of the recently reported murine and human EMAP II cytokine implicated in a variety of inflammatory disorders. The sequence of several proEMAP II proteins suggests that the p43 component of the complex is the precursor of the active mature cytokine after cleavage at a conserved Asp residue. The COOH-terminal moiety of p43 is also homologous to polypeptide domains found in bacterial methionyl- or phenylalanyl-tRNA synthetases and in the yeast Arc1p/G4p1 protein that associates with yeast methionyl-tRNA synthetase. Our results implicate the COOH-terminal moiety of p43 as a RNA binding domain. In the native state, as a component of the multisynthetase complex, p43 may be required for tRNA channeling and, after proteolytic processing occurring in tumor cells, would acquire inflammatory properties possibly related to apoptosis. The release of a truncated p43 from the complex could be involved in mediation of the signaling of tumor cells and stimulation of an acute inflammatory response. PMID- 9405473 TI - Reaction mechanism of (6-4) photolyase. AB - The (6-4) photolyase catalyzes the photoreversal of the (6-4) dipyrimidine photoproducts induced in DNA by ultraviolet light. Using the cloned Drosophila melanogaster (6-4) photolyase gene, we overproduced and purified the recombinant enzyme. The binding and catalytic properties of the enzyme were investigated using natural substrates, T[6-4]T and T[6-4]C, and the Dewar isomer of (6-4) photoproduct and substrate analogs s5T[6-4]T/thietane, mes5T[6-4]T, and the N methyl-3'T thietane analog of the oxetane intermediate. The enzyme binds to the natural substrates and to mes5T[6-4]T with high affinity (KD approximately 10(-9) 10(-10) M) and produces a DNase I footprint of about 20 base pairs around the photolesion. Several lines of evidence suggest that upon binding by the enzyme, the photoproduct flips out of the duplex. Of the four substrates that bind with high affinity to the enzyme, T[6-4]T and T[6-4]C are repaired with relatively high quantum yields compared with the Dewar isomer and the mes5T[6-4]T which are repaired with 300-400-fold lower quantum yield than the former two photoproducts. Reduction of the FAD cofactor with dithionite increases the quantum yield of repair. Taken together, the data are consistent with photoinduced electron transfer from reduced FAD to substrate, in a manner analogous to the cyclobutane pyrimidine dimer photolyase. PMID- 9405474 TI - Binding and catalytic properties of Xenopus (6-4) photolyase. AB - Xenopus (6-4) photolyase binds with high affinity to DNA bearing a (6-4) photoproduct and repairs it in a light-dependent reaction. To clarify its repair mechanism of (6-4) photolyase, we determined its binding and catalytic properties using synthetic DNA substrate which carries a photoproduct at a single location. The (6-4) photolyase binds to T[6-4]T in double-stranded DNA with high affinity (KD = 10(-9)) and to T[6-4]T in single-stranded DNA and T[Dewar]T in double- and single-stranded DNA although with slightly lower affinity (KD = approximately 2 x 10(-8)). Majority of the T[6-4]T-(6-4) photolyase complex dissociates very slowly (koff = 2.9 x 10(-5) s-1). Its absolute action spectrum without a second chromophore in the 350-600 nm region closely matches the absorption spectrum of the enzyme. The quantum yield (phi) of repair is approximately 0.11. The fully reduced form (E-FADH-) of (6-4) photolyase is catalytically active. Direct analysis of the photoreactivated product showed that (6-4) photolyase restores the original pyrimidines. These findings demonstrate that cis, syn-cyclobutane pyrimidine dimer photolyase and (6-4) photolyase are quite similar, but they are different with regard to the binding properties. PMID- 9405475 TI - Proteolytic cleavage of the beta1 subunit of platelet alpha2beta1 integrin by the metalloproteinase jararhagin compromises collagen-stimulated phosphorylation of pp72. AB - Early signaling events in the stimulation of platelets by collagen include the tyrosine phosphorylations of FcR gamma-chain, pp72(syk) and phospholipase Cgamma2. These events are dependent on the main platelet collagen receptor, alpha2beta1 integrin (glycoprotein Ia-IIa complex). We recently found that jararhagin, a 52-kDa snake venom metalloproteinase, selectively inhibits collagen induced platelet secretion and aggregation in parallel with the cleavage of the beta1 subunit of the alpha2beta1 integrin. The present study demonstrates that jararhagin also interferes with collagen-induced phosphorylation of the protein tyrosine kinase pp72(syk). This effect is not observed when the platelet aggregation response to collagen is inhibited by two venom RGD-containing disintegrins, contortrostatin and echistatin. These disintegrins inhibit platelet aggregation through their high affinity binding to the platelet alphaIIbbeta3 integrin (glycoprotein IIb-IIIa complex). We also show that mild stimulation by ADP of jararhagin-treated platelets, but not of platelets treated with the RGD containing disintegrins, restores the collagen-induced platelet aggregation. ADP also restored both pp72(syk) and pleckstrin phosphorylation of jararhagin-treated platelets in response to collagen, presumably via interaction of collagen with ADP-activated alphaIIbbeta3 integrin. Thus, RGD-containing disintegrins do not interfere with agonist-induced pp72(syk) phosphorylation but inhibit aggregation through occupancy of the alphaIIbbeta3 integrin. Conversely, jararhagin affects early platelet signaling events in response to collagen through its effects on the alpha2beta1 integrin without interfering with the function of the alphaIIbbeta3 integrin. Our demonstration that the degradation of the beta1 subunit of alpha2beta1 by jararhagin results in the loss of pp72(syk) phosphorylation, suggests that this subunit is critically involved in collagen induced platelet signaling. PMID- 9405476 TI - Activation of nuclear transcription factor NF-kappaB by interleukin-1 is accompanied by casein kinase II-mediated phosphorylation of the p65 subunit. AB - In fibroblasts and hepatoma cells, interleukin-1 (IL-1) treatment results in the rapid nuclear accumulation of the transcription factor NF-kappaB, present largely as p65 (RelA)/p50 heterodimers. It is well established that this process is dependent in large part upon the phosphorylation and subsequent degradation of the cytosolic inhibitor IkappaB. We looked for other IL-1-induced modifications of NF-kappaB components and found that, in both cell types, IL-1 stimulation led, within minutes, to phosphorylation of both NF-kappaB p65 and p50. Phosphorylation of p65 was sustained for at least 30 min after addition of the cytokine and occurred principally upon serine residues. Immunoprecipitates of NF-kappaB complexes contained an associated protein kinase, the biochemical characteristics of which were indistinguishable from casein kinase II (CKII). Purified CKII efficiently phosphorylated p65 in vitro, apparently on the same major sites that became phosphorylated in intact IL-1-treated cells. Although IL-1 treatment caused little apparent stimulation of total cellular CKII activity, the fraction that was specifically associated with NF-kappaB complexes was markedly elevated by the cytokine. The association of CKII with NF-kappaB occurred in the cytoplasm, suggesting that this phosphorylation might be involved either in control of translocation of the activated complex or in modulation of its DNA binding properties. PMID- 9405477 TI - Identification of an endothelial cell-specific regulatory region in the murine endothelin-1 gene. AB - Endothelin-1 is a 21-amino acid peptide first characterized as a potent vasoactive compound synthesized by endothelial cells. Because of its high level cell-restricted pattern of expression, we have employed this gene as a model for investigating the DNA and protein elements that mediate endothelial cell-specific gene expression. In this study we have identified a complex positive regulatory region located at base pairs -364 to -320 in the murine endothelin-1 gene. This region consists of three functionally dependent elements, ETE-C, ETE-D, and ETE E, which are all required for full activity. When a 43-base pair fragment containing these three elements was employed in heterologous promoter experiments, this sequence was capable of increasing transcriptional activity in an endothelial cell-specific fashion. None of the elements contains a recognized consensus sequence known to bind transcriptional regulatory proteins in higher eukaryotes; however, each element does appear to mediate protein binding. The combination of all three elements promotes binding of a protein complex that is endothelial cell-specific. This is the first evidence for an endothelial cell specific DNA regulatory element and cognate binding proteins. PMID- 9405478 TI - Tough tendons. Mussel byssus has collagen with silk-like domains. AB - The primary structure of the alpha-chain of preCol-D (molecular mass = 80 kDa), a tanned collagenous protein predominating in the distal portion of the byssal threads of the mussel Mytilus edulis, was deduced from cDNA to encode an unprecedented natural block copolymer with three major domain types: a central collagen domain flanked by fibroin-like domains and followed by histidine-rich termini. The fibroin-like domains have sequence motifs that strongly resemble the crystalline polyalanine-rich and amorphous glycine-rich regions of spider dragline silk fibroins. The terminal regions resemble the histidine-rich domains of a variety of metal-binding proteins. The silk domains may toughen the collagen by increasing its strength and extensibility. PreCol-D expression is limited to the mussel foot, which contains a longitudinal gradient of preCol-D mRNA. This gradient increases linearly in the proximal to distal direction and reaches a maximum just before the distal depression of the foot. PMID- 9405479 TI - Tissue-specific distribution and modulatory role of the gamma subunit of the Na,K ATPase. AB - The Na,K-ATPase comprises a catalytic alpha subunit and a glycosylated beta subunit. Another membrane polypeptide, gamma, first described by Forbush et al. (Forbush, B., III, Kaplan, J. H., and Hoffman, J. F. (1978) Biochemistry 17, 3667 3676) associates with alpha and beta in purified kidney enzyme preparations. In this study, we have used a polyclonal anti-gamma antiserum to define the tissue specificity and topology of gamma and to address the question of whether gamma has a functional role. The trypsin sensitivity of the amino terminus of the gamma subunit in intact right-side-out pig kidney microsomes has confirmed that it is a type I membrane protein with an extracellular amino terminus. Western blot analysis shows that gamma subunit protein is present only in membranes from kidney tubules (rat, dog, pig) and not those from axolemma, heart, red blood cells, kidney glomeruli, cultured glomerular cells, alpha1-transfected HeLa cells, all derived from the same (rat) species, nor from three cultured cell lines derived from tubules of the kidney, namely NRK-52E (rat), LLC-PK (pig), or MDCK (dog). To gain insight into gamma function, the effects of the anti-gamma serum on the kinetic behavior of rat kidney sodium pumps was examined. The following evidence suggests that gamma stabilizes E1 conformation(s) of the enzyme and that anti-gamma counteracts this effect: (i) anti-gamma inhibits Na,K ATPase, and the inhibition increases at acidic pH under which condition the E2(K) --> E1 phase of the reaction sequence becomes more rate-limiting, (ii) the oligomycin-stimulated increase in the level of phosphoenzyme was greater in the presence of anti-gamma indicating that the antibody shifts the E1 left and right arrow left and right arrow E2P equilibria toward E2P, and (iii) when the Na+ ATPase reaction is assayed with the Na+ concentration reduced to levels ( --> E2P transition, anti-gamma is stimulatory. These observations taken together with evidence that the pig gamma subunit, which migrates as a doublet on polyacrylamide gels, is sensitive to digestion by trypsin, and that Rb+ ions partially protect it against this effect, indicate that the gamma subunit is a tissue-specific regulator which shifts the steady state equilibria toward E1. Accordingly, binding of anti-gamma disrupts alphabeta gamma interactions and counteracts these modulatory effects of the gamma subunit. PMID- 9405481 TI - A novel regulatory mechanism in the mitogen-activated protein (MAP) kinase cascade. Role of nuclear export signal of MAP kinase kinase. AB - Mitogen-activated protein kinase (MAPK) kinase (MAPKK, also known as MEK), a direct activator for MAPK/extracellular signal-regulated kinase, localizes to the cytoplasm excluded from the nucleus during signal transmission. This nuclear exclusion of MAPKK is directed by its nuclear export signal (NES), but its physiological significance has been unknown. We have found that disruption of the NES dramatically potentiates the ability of constitutively active MAPKK to induce morphological changes and malignant transformation of fibroblastic cells. Readdition of the NES sequence reversed the effects induced by the NES disruption. Moreover, we observed that a dramatic increase of activated MAPK in the nucleus was induced by the NES-disrupted MAPKK and that coexpression of MAPK phosphatase-1 (CL-100) or a kinase negative form of MAPK counteracted the phenotypes induced by the NES-disrupted MAPKK, indicating the crucial role of MAPK in the responses. These findings reveal a novel regulatory role of the NES of MAPKK that may be essential for proper signal transductions. PMID- 9405480 TI - On the role of Arg-210 and Glu-219 of subunit a in proton translocation by the Escherichia coli F0F1-ATP synthase. AB - A strain of Escherichia coli was constructed which had a complete deletion of the chromosomal uncB gene encoding subunit a of the F0F1-ATP synthase. Gene replacement was facilitated by a selection protocol that utilized the sacB gene of Bacillus subtilis cloned in a kanamycin resistance cartridge (Ried, J. L., and Collmer, A. (1987) Gene (Amst.) 57, 239-246). F0 subunits b and c inserted normally into the membrane in the DeltauncB strain. This observation confirms a previous report (Hermolin, J., and Fillingame, R. H. (1995) J. Biol. Chem. 270, 2815-2817) that subunit a is not required for the insertion of subunits b and c. The DeltauncB strain has been used to characterize mutations in Arg-210 and Glu 219 of subunit a, residues previously postulated to be essential in proton translocation. The aE219G and aE219K mutants grew on a succinate carbon source via oxidative phosphorylation and membranes from these mutants exhibited ATPase coupled proton translocation (i.e. ATP driven 9-amino-6-chloromethoxyacridine quenching responses that were 60-80% of wild type membranes). We conclude that the aGlu-219 residue cannot play a critical role in proton translocation. The aR210A mutant did not grow on succinate and membranes exhibited no ATPase-coupled proton translocation. However, on removal of F1 from membrane, the aR210A mutant F0 was active in passive proton translocation, i.e. in dissipating the DeltapH normally established by NADH oxidation with these membrane vesicles. aR210A membranes with F1 bound were also proton permeable. Arg-210 of subunit a may play a critical role in active H+ transport that is coupled to ATP synthesis or hydrolysis, but is not essential for the translocation of protons across the membranes. PMID- 9405482 TI - BCR/ABL-induced leukemogenesis causes phosphorylation of Hef1 and its association with Crkl. AB - BCR/ABL is considered responsible for the development of Philadelphia chromosome positive leukemia. Experimental animal models, such as transgenic mice, have demonstrated unambiguously that Bcr/Abl is capable of inducing leukemogenesis. The adaptor molecule Crkl is a major in vivo substrate of the deregulated Bcr/Abl tyrosine kinase and functions as a molecular link with other signaling proteins. While associated in vivo with Bcr/Abl through its SH3 domain, Crkl can interact simultaneously via its SH2 domain with other tyrosine-phosphorylated proteins. Here we report the identification of prominently tyrosine-phosphorylated proteins with a molecular mass of approximately 110 kDa, which bind specifically to the Crkl SH2 domain in leukemic tissues of P190BCR/ABL transgenic mice. We demonstrate that these proteins are identical to Hef1/Cas-L, which is related to p130(Cas). The proto-oncoprotein p120(Cbl) and Hef1, but not p130(Cas), were detectably phosphorylated on tyrosine in P190Bcr/Abl-expressing leukemic cells and were found in complex with Crkl, showing the existence of protein complexes in P190Bcr/Abl leukemic cells, consisting of P190Bcr/Abl, Crkl, and Hef1 or p120(Cbl). This supports a model in which Crkl acts as mediator between Bcr/Abl and downstream effectors. Since Hef1 is involved in the beta1-integrin signaling pathway, our study demonstrates that Bcr/Abl could specifically interfere with normal beta1-integrin signaling. PMID- 9405483 TI - Phosphorylation of steroidogenic acute regulatory protein (StAR) modulates its steroidogenic activity. AB - Steroidogenic acute regulatory protein (StAR) plays a critical role in steroid hormone synthesis. StAR is thought to increase the delivery of cholesterol to the inner mitochondrial membrane where P450scc resides. Tropic hormones acting through the intermediacy of cAMP rapidly increase pregnenolone synthesis, and this rapid steroidogenic response is believed to be due to StAR's action. The StAR protein contains two consensus sequences for phosphorylation catalyzed by protein kinase A that are conserved across all species in which the amino acid sequence of the StAR protein has been determined. We demonstrated that human StAR expressed in COS-1 cells exists in at least four species detectable by two dimensional gel electrophoresis followed by Western blotting. The two more acidic species disappeared after treatment of the cell extracts with alkaline phosphatase. 32P was incorporated into StAR protein immunoprecipitated from COS-1 cell extracts, and a 10-min treatment with 8-bromo-cAMP increased 32P incorporation into the StAR preprotein. StAR protein generated by in vitro transcription/translation was phosphorylated by the protein kinase A catalytic subunit in the presence of [gamma-32P]ATP. Mutation of potential sites for protein kinase A-mediated phosphorylation at serine 57 and serine 195 to alanines, individually, reduced 32P incorporation from labeled ATP into StAR preprotein produced by in vitro transcription/translation when incubated with protein kinase A catalytic subunit. 32P labeling of StAR protein expressed in COS 1 cells was also reduced when serine 57 or serine 195 were mutated to alanines. A double mutant in which both serine 57 and serine 195 were changed to alanines displayed markedly reduced 32P incorporation. To determine the functional significance of StAR phosphorylation, we tested the steroidogenic activity of the wild-type StAR and mutated StAR proteins in COS-1 cells expressing the human cholesterol side chain cleavage enzyme system. Mutation of the conserved protein kinase A phosphorylation site at serine 57 had no effect on pregnenolone synthesis. However, mutation of the serine residue at 195 resulted in an approximately 50% reduction in pregnenolone production. The S195A mutant construct did not yield the more acidic species of StAR detected in two dimensional Western blots, indicating that the mutation affected the ability of the protein to be post-translationally modified. Mutation of the corresponding serine residues in murine StAR (Ser56 and Ser194) to alanines yielded results that were similar to those obtained with human StAR; the S56A mutant displayed a modest reduction in steroidogenic activity, whereas the S194A mutant had approximately 40% of the activity of murine wild-type StAR. In contrast to the human S195A mutation, conversion of serine 195 to an aspartic acid residue had no effect on steroidogenic activity, consistent with the idea that a negative charge at this site modulates StAR function. Our observations suggest that phosphorylation of serine 194/195 increases the biological activity of StAR and that this post- or co-translational event accounts, in part, for the immediate effects of cAMP on steroid production. PMID- 9405484 TI - Yeast Gal11 and transcription factor IIE function through a common pathway in transcriptional regulation. AB - The global transcription regulator Gal11, a component of RNA polymerase II holoenzyme, is required for full expression of many genes in yeast. We previously reported that Gal11 binds the small (Tfa2) and large (Tfa1) subunits of the general transcription factor (TF) IIE through Gal11 functional domains A and B, respectively. Here we demonstrate that the C-terminal basic region in Tfa2 is responsible for binding to domain A, whereas both the N-terminal hydrophobic and internal glutamic acid-rich regions in Tfa1 are responsible for binding to domain B. Yeast cells bearing a C-terminal deletion encompassing the Gal11-interacting region in each of the two TFIIE subunits, being viable, exhibited no obvious phenotype. In contrast, combination of the two deletions (TFIIE-DeltaC) showed phenotypes similar to those of gal11 null mutations. The levels of mRNA from TATA containing genes, but not from TATA-less genes, decreased in TFIIE-DeltaC to an extent comparable to that in the gal11 null mutant. Combination of TFIIE-DeltaC with a gal11 null mutation did not result in an enhanced effect, suggesting that both TFIIE and Gal11 act in a common regulatory pathway. In a reconstituted cell free system, Gal11 protein stimulated basal transcription in the presence of wild type TFIIE. Such a stimulation was not seen in the presence of TFIIE-DeltaC. PMID- 9405485 TI - The platelet-derived growth factor beta receptor triggers multiple cytoplasmic signaling cascades that arrive at the nucleus as distinguishable inputs. AB - Stimulation of the platelet-derived growth factor beta receptor (betaPDGFR) activates enzymes such as phosphatidylinositol 3-kinase (PI3K) and phospholipase Cgamma1 (PLCgamma), which ultimately initiate nuclear responses such as enhanced expression of immediate early genes. In an attempt to compare the signaling cascades initiated by PI3K and PLCgamma, we examined the activation of a panel of immediate early genes by betaPDGFR mutants, which preferentially engage PI3K or PLCgamma. When expressed in A431 cells, the wild type receptor and to a lesser extent the mutant receptor that associates with PLCgamma (Y1021) was able to up regulate c-fos, junB, and KC mRNA expression. In contrast, the receptor mutant that engages PI3K (Y740/51) poorly stimulated c-fos mRNA expression and did not significantly stimulate expression of either JunB or KC. Receptor mutants that did not associate with either PI3K or PLCgamma were dramatically compromised or unable to increase expression of any of these immediate early genes. The differential ability of the Y1021 and Y740/51 receptors to activate c-fos correlated well with an apparent difference in their ability to engage distinct protein kinase C family members. However there did appear to be a degree of redundancy in the cytoplasmic signaling pathways initiated by PI3K and PLCgamma, since both the Y1021 and Y740/51 receptors were able to activate an AP-1 responsive element. We conclude that recruitment of signal relay enzymes to the betaPDGFR is necessary for PDGF-dependent activation of at least some immediate early genes. In addition, whereas the betaPDGFR activates multiple signaling enzymes capable of activating the same nuclear response (activation of c-fos), these signaling cascades do not appear to converge in the cytoplasm but arrive at the nucleus as distinguishable inputs. PMID- 9405486 TI - Enzyme-substrate intermediate at a specific lysine residue is required for deoxyhypusine synthesis. The role of Lys329 in human deoxyhypusine synthase. AB - Deoxyhypusine synthase catalyzes the first step in the post-translational synthesis of hypusine [Nepsilon-(4-amino-2-hydroxybutyl)lysine] in eukaryotic translation initiation factor 5A. We recently reported biochemical evidence for a covalent enzyme-substrate intermediate involving a specific lysine residue (Lys329) in human deoxyhypusine synthase (Wolff, E. C., Folk, J. E., and Park, M. H. (1997) J. Biol. Chem. 272, 15865-15871). In an effort to evaluate the role of this enzyme-substrate intermediate in catalysis, we carried out site-directed mutagenesis (Lys to Arg and/or Ala) of the conserved lysine residues in human deoxyhypusine synthase. A drastic reduction in enzyme intermediate formation and enzymatic activities was observed with mutant proteins with substitution at Lys287 but not with those with mutations at residues 141, 156, 205, 212, 226, 251, or 338. Lys to Ala or Lys to Arg substitution at Lys329 totally abolished covalent enzyme-substrate intermediate formation and deoxyhypusine synthesis activity, indicating that Lys329 is the unique site for the enzyme intermediate and that it is absolutely required for deoxyhypusine synthesis in the eukaryotic translation initiation factor 5A precursor. The K329A mutant showed spermidine cleavage activity ( approximately 6% of the wild type enzyme) suggesting that in contrast to deoxyhypusine synthesis, spermidine cleavage can occur without enzyme intermediate formation. PMID- 9405487 TI - Divergent signaling capacities of the long and short isoforms of the leptin receptor. AB - Leptin receptors include a long form (OBRl) with 302 cytoplasmic residues that is presumed to mediate most or all of leptins signaling, and several short forms, including one (OBRs) that has 34 cytoplasmic residues, is widely expressed, and is presumed not to signal but to mediate transport or clearance of leptin. We studied the abilities of these two receptor isoforms to mediate signaling in transfected cells. In response to leptin, OBRl, but not OBRs, underwent tyrosine phosphorylation that was enhanced by co-expression with JAK2. In cells expressing receptors and JAK2, both OBRs and OBRl mediated leptin-dependent tyrosine phosphorylation of JAK2, and this was abolished with OBRs when the Box 1 motif was mutated. In cells expressing receptors, JAK2 and IRS-1, leptin induced tyrosine phosphorylation of IRS-1 through OBRs and OBRl. In COS cells expressing hemagglutinin-ERK1 and receptors, leptin increased ERK1 kinase activity through OBRl, with the magnitude increased by co-expression of JAK1 or JAK2, and to a lesser degree through OBRs, despite greater receptor expression. In stable Chinese hamster ovary cell lines expressing OBRs or OBRl, leptin stimulated endogenous ERK2 phosphorylation. Whereas leptin stimulated tyrosine phosphorylation of hemagglutinin-STAT3 and induction of a c-fos luciferase reporter plasmid through OBRl, OBRs was without effect in these assays. In conclusion, OBRl is capable of signaling to IRS-1 and mitogen-activated protein kinase via JAK, in addition to activating STAT pathways. Although substantially weaker than OBRl, OBRs is capable of mediating signal transduction via JAK, but these activities are of as yet unknown significance for leptin biology in vivo. PMID- 9405488 TI - The DNA dependence of the ATPase activity of human DNA topoisomerase IIalpha. AB - We have purified human topoisomerase IIalpha from HeLa cells and studied its ATPase reaction. The ATPase activity is stimulated by DNA and shows apparent Michaelis-Menten kinetics. Although the ATPase activity of human topoisomerase IIalpha is lower than that of Saccharomyces cerevisiae, it is more active in decatenation, implying more efficient coupling of the ATPase to DNA strand passage under these conditions. Using plasmid pBR322 as the DNA cofactor, the reaction shows hyperstimulation by DNA at a base pair to enzyme dimer ratio of 100-200:1. When DNA fragments are used as the cofactor, the reaction requires > approximately 100 base pairs to stimulate the activity and fragments of approximately 300 base pairs show hyperstimulation. This behavior can be rationalized in terms of the enzyme requiring fragments that can bind to both the DNA gate and the ATP-operated clamp in order for the ATPase reaction to be stimulated. Hyperstimulation is a consequence of the saturation of DNA with enzyme. The mechanistic implications of these results are discussed. PMID- 9405489 TI - Isoform-specific activation and structural diversity of calmodulin kinase I. AB - We earlier confirmed that there are isoforms of Ca2+/calmodulin (CaM)-dependent protein kinase I (CaM kinase I) (CaM kinase Ibeta1 and Igamma) beside CaM kinase Ialpha by cDNA cloning (Yokokura, H., Terada, O., Naito, Y., and Hidaka, H. (1997) Biochim. Biophys. Acta 1338, 8-12). Here, we demonstrate the existence of an isoform-specific activation mechanism of CaM kinase I and alternative splicing specifically regulating CaM kinase I (CaM kinase Ibeta2) in the central nervous system. To cast light on isoform structure-enzyme activity relationships, CaM kinase Ibeta1, Ibeta2, and Ialpha were expressed separately using a baculovirus/Sf9 cell expression system. The novel CaM kinase Ibeta2 isoform demonstrated similar catalytic activity to those of CaM kinase Ibeta1 and Ialpha. Interestingly, CaM kinase Ibeta1 and Ibeta2 both can activate CaM kinase Ialpha activity via phosphorylation at Thr177. Reverse transcribed-polymerase chain reaction analysis showed that CaM kinase Ibeta2 is dominant in the cerebrum and cerebellum, whereas CaM kinase Ibeta1 is present in peripheral tissues such as liver, heart, lung, kidney, spleen, and testis. CaM kinase Ibeta2 was also detected with an anti-CaM kinase Ibeta2 antibody in PC12 cells. The results indicate that alternative splicing is a means for tissue-specific expression of CaM kinase Ibeta. Thus the Thr177 residue of CaM kinase Ialpha is phosphorylated by not only CaM kinase kinase but also CaM kinase Ibeta for activation of the enzyme. PMID- 9405490 TI - Khafrefungin, a novel inhibitor of sphingolipid synthesis. AB - In the course of screening for antifungal agents we have discovered a novel compound isolated from an endophytic fungus that inhibits fungal sphingolipid synthesis. Khafrefungin, which is composed of aldonic acid linked via an ester to a C22 modified alkyl chain, has fungicidal activity against Candida albicans, Cryptococcus neoformans, and Saccharomyces cerevisiae. Sphingolipid synthesis is inhibited in these organisms at the step in which phosphoinositol is transferred to ceramide, resulting in accumulation of ceramide and loss of all of the complex sphingolipids. In vitro, khafrefungin inhibits the inositol phosphoceramide synthase of C. albicans with an IC50 of 0.6 nM. Khafrefungin does not inhibit the synthesis of mammalian sphingolipids thus making this the first reported compound that is specific for the fungal pathway. PMID- 9405491 TI - Agrin is a major heparan sulfate proteoglycan in the human glomerular basement membrane. AB - Agrin is a heparan sulfate proteoglycan (HSPG) that is highly concentrated in the synaptic basal lamina at the neuromuscular junction (NMJ). Agrin-like immunoreactivity is also detected outside the NMJ. Here we show that agrin is a major HSPG component of the human glomerular basement membrane (GBM). This is in addition to perlecan, a previously characterized HSPG of basement membranes. Antibodies against agrin and against an unidentified GBM HSPG produced a strong staining of the GBM and the NMJ, different from that observed with anti-perlecan antibodies. In addition, anti-agrin antisera recognized purified GBM HSPG and competed with an anti-GBM HSPG monoclonal antibody in ELISA. Furthermore, both antibodies recognized a molecule that migrated in SDS-PAGE as a smear and had a molecular mass of approximately 200-210 kD after deglycosylation. In immunoelectron microscopy, agrin showed a linear distribution along the GBM and was present throughout the width of the GBM. This was again different from perlecan, which was exclusively present on the endothelial side of the GBM and was distributed in a nonlinear manner. Quantitative ELISA showed that, compared with perlecan, the agrin-like GBM HSPG showed a sixfold higher molarity in crude glomerular extract. These results show that agrin is a major component of the GBM, indicating that it may play a role in renal ultrafiltration and cell matrix interaction. (J Histochem Cytochem 46:19-27, 1998) PMID- 9405492 TI - Oncomodulin is expressed exclusively by outer hair cells in the organ of Corti. AB - Oncomodulin (OM) is a small, acidic calcium-binding protein first discovered in a rat hepatoma and later found in placental cytotrophoblasts, the pre-implantation embryo, and in a wide variety of neoplastic tissues. OM was considered to be exclusively an oncofetal protein until its recent detection in extracts of the adult guinea pig's organ of Corti. Here we report that light and electron microscopic immunostaining of gerbil, rat, and mouse inner ears with a monoclonal antibody against recombinant rat OM localizes the protein exclusively in cochlear outer hair cells (OHCs). At the ultrastructural level, high gold labeling density was seen overlying the nucleus, cytoplasm, and the cuticular plate of gerbil OHCs. Few, if any, gold particles were present over intracellular organelles and the stereocilia. Staining of a wide range of similarly processed gerbil organs failed to detect immunoreactive OM in any other adult tissues. The mammalian genome encodes one alpha- and one beta-isoform of parvalbumin (PV). The widely distributed alpha PV exhibits a very high affinity for Ca2+ and is believed to serve as a Ca2+ buffer. By contrast, OM, the mammalian beta PV, displays a highly attenuated affinity for Ca2+, consistent with a Ca2+-dependent regulatory function. The exclusive association of OM with cochlear OHCs in mature tissues is likely to have functional relevance. Teleological considerations favor its involvement in regulating some aspect of OHC electromotility. Although the fast electromotile response of OHCs does not require Ca2+, its gain and magnitude are modulated by efferent innervation. Therefore, OM may be involved in mediation of intracellular responses to cholinergic stimulation, which are known to be Ca2+ regulated. (J Histochem Cytochem 46:29-39, 1998) PMID- 9405493 TI - p53 expression in human carcinomas: could flow cytometry be an alternative to immunohistochemistry? AB - Several studies have shown that p53 expression has important clinical implications as an indicator of prognosis and response to chemotherapy or radiotherapy in different human tumor types. Determination of p53 expression by immunohistochemistry (IHC) has been incorporated into routine practice and its reliability has been consolidated. However, flow cytometric (FCM) analysis might represent an important objective and rapid approach. In the present study we determined p53 expression by IHC and FCM on a series of 118 human solid tumors. IHC determination was performed on histological sections and FCM analysis on cell suspensions. Low correlation coefficients (rs from 0.22 to 0.57) were observed between IHC and FCM data from individual tumors. By considering the IHC approach as the gold standard, high sensitivity and low specificity were found for FCM in detecting p53 expression. The FCM analysis of p53 expression and DNA content showed p53-positive cells in all cell cycle phases. Moreover, in most breast, lung, and colon aneuploid tumors (77%), p53-positive cells were detected only in the subpopulations with abnormal DNA content. In conclusion, FCM-p53 expression cannot be used alternatively to IHC determination, and its clinical relevance remains to be validated. Nevertheless, FCM may provide important information about p53 protein expression in the different subpopulations and cell cycle phases. (J Histochem Cytochem 46:41-47, 1998) PMID- 9405494 TI - Effects of c-mpl ligand on cytoplasmic maturation of murine megakaryocytes and on platelet production. AB - To test the hypothesis that the c-mpl ligand is not a primary factor in thrombocytopoiesis, we investigated the biological effects of recombinant human (rh) c-mpl ligand on differentiation of murine progenitor cells and on maturation of the cultured murine megakaryocytes under serum-free conditions on the basis of ploidy distribution, megakaryocyte/platelet-specific surface antigen CD 61 [glycoprotein (GP) IIIa], and cytoplasmic acetylcholinesterase (AchE) expression in vitro. In addition, we studied the effect of c-mpl ligand on proplatelet formation (PPF) by murine mature megakaryocytes. AchE was less strongly expressed in cultured megakaryocytic cells stimulated by c-mpl ligand than in those stimulated by recombinant murine (rm) IL-3 + rh IL-6 during the differentiation of progenitor cells. Less CD 61 was expressed by c-mpl ligand during both the differentiation of progenitor cells and the maturation of megakaryocytes compared with that by rm IL-3 + rh IL-6. Endomitosis, however, was more stimulated by c mpl ligand than by rm IL-3 + rh IL-6 under both conditions. Furthermore, PPF of mature megakaryocytes was not stimulated by c-mpl ligand. These results indicate that c-mpl ligand stimulates the nuclear development of megakaryocytic cells but that it does not stimulate cytoplasmic maturation and PPF as much as IL-6. These data strongly suggest that c-mpl ligand is not a primary factor in platelet pro duction. (J Histochem Cytochem 46:49-57, 1998) PMID- 9405495 TI - Distribution of H type 1 and of H type 2 antigens of ABO blood group in different cells of human submandibular gland. AB - We have examined the immunohistochemical distribution of H Type 1 and of H Type 2 substances of the ABO blood group system in human submandibular gland using either of the two anti-H monoclonal antibodies MAb 1E3 and MAb 3A5. MAb 3A5 was specific for H Type 2, and MAb 1E3 reacted with each of H Type 1-H Type 4 artificial antigens. We have developed a competitive inhibition method against H Type 2 and have obtained MAb 1E3, which is fairly specific for H Type 1 under certain conditions. Mucous cells from secretors were strongly stained by 1E3 and weakly by 3A5, whereas those from nonsecretors showed no reaction with 1E3 and 3A5. Serous cells from both secretors and nonsecretors were stained neither by 1E3 nor by 3A5. Striated and interlobular duct cells were strongly stained by 1E3 and by 3A5, regardless of the secretor status. These results indicated that the expressions of the H Type 1 and H Type 2 in different cell types of the submandibular gland were controlled by different genes. In addition, we have determined the acceptor specificity of two alpha(1,2)fucosyltransferases (H and Se enzymes) after transient expressions of the FUT1 and FUT2 in COS7 cells, and found that the H enzyme activity was similar for both Type 1 and Type 2 precursors, and that Se enzyme activity with the Type 1 precursor was higher than that with the Type 2 precursor. Expression of the H Type 1 antigen in mucous cells was found to be dependent on the Se gene, whereas expressions of the H Type 1 and H Type 2 antigens in striated and interlobular duct cells were dependent on the H gene. (J Histochem Cytochem 46:69-76, 1998) PMID- 9405496 TI - Expression of prostaglandin G/H synthase type 1, but not type 2, in human ovarian adenocarcinomas. AB - Prostaglandin endoperoxide synthase (PGHS) is a key rate-limiting enzyme in prostaglandin biosynthesis. PGHS has recently been shown to be expressed in human colorectal cancers and in experimental cutaneous papillomas and carcinomas. However, PGHS expression has not been investigated in ovarian cancers. The objectives of this study were to determine whether PGHS isoenzymes are expressed in human ovarian cancer and, if so, to identify which isoform is involved (PGHS-1 and/or PGHS-2) and to characterize its cellular localization. Sixteen human ovarian adenocarcinomas were studied by immunohistochemistry using specific antibodies recognizing PGHS-1 or PGHS-2. Results showed that all adenocarcinomas demonstrated the presence of tumor cells expressing PGHS-1 but not PGHS-2. Patterns of staining of tumor cells varied among different types of adenocarcinomas, with cells presenting either a mostly diffuse cytoplasmic immunoreactivity or, alternatively, a staining mainly concentrated around the nucleus. No correlation between the intensity of the immunostaining and the degree of malignancy of tumors could be established (r 5 20.03; p>0.05). Immunoblot analysis with PGHS-1-selective antibodies of cell extracts from adenocarcinomas revealed the presence of a characteristic 72,000 Mr immunoreactive band. Therefore, these results show for the first time that PGHS-1 is expressed in human ovarian adenocarcinomas. (J Histochem Cytochem 46:77-84, 1998) PMID- 9405497 TI - Comparison of two methods of staining apoptotic cells of leukemia cell lines. Terminal deoxynucleotidyl transferase and DNA polymerase I reactions. AB - We compared two methods to stain apoptotic cells, one using terminal deoxynucleotidyl transferase (TDT), the other DNA polymerase I, using leukemia cell lines treated with anti-Fas monoclonal antibody (MAb). Both TDT and polymerase I strongly reacted with fragmented nuclei of apoptotic MOLT-16 and Jurkat cells, but only polymerase I strongly reacted with nonfragmented nuclei of early apoptotic cells. Anti-Fas MAb-treated MOLT-4 cells showed morphological changes corresponding to early apoptosis and were strongly positive for polymerase I only. MOLT-16 and Jurkat cells treated with anti-Fas MAb and inhibitors of endonuclease and poly(ADP-ribose) polymerase showed the morphology of early apoptosis but were not strongly stained by TDT. Because DNA polymerase I has nick-translation activity, it is possible that DNA polymerase I reaction is positive in early apoptotic cells by detecting single-strand DNA cleavage, which occurs before extensive oligonucleosomal DNA cleavage and late morphological changes of apoptosis in leukemia cell lines. Although TDT is widely used to stain apoptotic cells, DNA polymerase I may be more applicable in special cases of apoptosis, in which cells undergo single-strand rather than double-strand DNA breaks. However, the procedure has limitations, such as the necessity to use cell smears for comparison with the TDT reaction. (J Histochem Cytochem 46:85-90, 1998) PMID- 9405498 TI - Immunolocalization of vacuolar-type H+-ATPase in rat submandibular gland and adaptive changes induced by acid-base disturbances. AB - Using antibodies against the 31-kD and 70-kD subunits of vacuolar type H+-ATPase (V-ATPase) and light microscopic immunocytochemistry, we have demonstrated the presence of this V-ATPase in rat submandibular gland. We have also investigated the adaptive changes of this transporter during acid-base disturbances such as acute and chronic metabolic acidosis or alkalosis. Our results show intracellularly distributed V-ATPase in striated, granular, and main excretory duct cells in controls, but no V-ATPase immunoreaction in acinar cells. Both acute and chronic metabolic acidosis caused a shift in V-ATPase away from diffuse distribution towards apical localization in striated and granular duct cells, suggesting that a V-ATPase could be involved in the regulation of acid-base homeostasis. In contrast, during acidosis the main excretory duct cells showed no changes in the V-ATPase distribution compared to controls. With acute and chronic metabolic alkalosis, no changes in the V-ATPase distribution occurred. (J Histochem Cytochem 46:91-100, 1998) PMID- 9405499 TI - Immunocytochemical localization of the prohormone convertases PC1 and PC2 in rat prolactin cells. AB - The prohormone convertases PC1 and PC2 are subtilisin-related endopeptidases that process prohormone and neuropeptide precursors. Using different ultrastructural immunocytochemical approaches, we have investigated their intracellular distribution in a neuroendocrine cell type that has not been examined thus far, the rat anterior pituitary lactotrope. These cells secrete mainly prolactin and also express the neuroendocrine-specific protein secretogranin II, which is considered a peptide precursor. Our study provides evidence for the expression of PC1 and PC2 in rat lactotropes and provides new information on their subcellular localization. Apart from their presence in the secretory granules, PC1 and PC2 displayed different major localization along the secretory pathway. PC1 immunoreactivity was concentrated in the Golgi apparatus, whereas PC2 immunoreactivity was prominent in the rough endoplasmic reticulum (RER). These observations provide morphological support for previous biochemical analysis of proPC1 and proPC2 post-translational processing, which has demonstrated that PC1 exits very rapidly from the RER, whereas PC2 is retained much longer in this compartment. (J Histochem Cytochem 46:101-108, 1998) PMID- 9405500 TI - Nonspecific labeling of myelin with secondary antisera and high concentrations of Triton X-100. AB - Triton X-100 is used in immunohistochemistry to make tissue permeable, to present certain antigens to antisera, and to prevent certain nonspecific interactions. This detergent is routinely dissolved in buffers at concentrations of 0.01-0.2%. Using high concentrations of Triton X-100 (0.2-2%) and anti-immunoglobulins G (anti-IgGs), labeling of myelin and microglia was detected in fixed brain tissue by indirect fluorescence and avidin-biotin-immunoperoxidase techniques. Differences were found between the species studied (mouse and rat), the type of anti-IgG (anti-mouse, anti-rabbit, anti-sheep, anti-rat, or anti-guinea pig), the detergent concentration, and whether Triton X-100 was included in the incubation media or applied as a pretreatment. Mouse brain displayed strong myelin labeling with all anti-IgGs but rat brain only with anti-rabbit or anti-sheep IgGs. Staining of ramified microglia occurred only in mouse tissue when anti-mouse IgG was used. Nonspecific staining of myelin was also intense in paraffin-embedded tissue and in human brain frozen sections. These results are significant for the prevention of undesirable staining in routine immunolabeling and they also provide a comparatively inexpensive, easy to perform strong labeling of myelin. In addition, the double marker signal (peroxidase and fluorescence) is useful for double labeling studies. (J Histochem Cytochem 46:109-117, 1998) PMID- 9405501 TI - Integrin expression in developing smooth muscle cells. AB - We studied the specific expression patterns and distributions of alpha1 and beta1 integrin subunits, the major cell adhesion receptors in smooth muscle, in developing smooth muscle cells from 16-, 18-, and 20-day embryonic gizzards and from 1- and 7-day post hatch chick gizzards by SDS-PAGE, immunoblotting, and immunoelectron microscopy. Antibodies raised against alpha1 and beta1 integrins isolated from avian gizzards were used as probes. Gels and blots showed that the amount of alpha1 and beta1 integrins increased as age increased, with major increases at 1 and 7 days post hatch. Image analysis of immunoelectron micrographs demonstrated that statistically significant labeling increases occurred between embryonic Days 16 and 18, between embryonic Day 20 and 1 day post hatch, and between 1 day and 7 days post hatch. Immunolabeling with both anti-alpha1 and anti-beta1 integrin was prominent at membrane-associated dense plaques (MADPs) and at filament anchoring regions at cell ends. This indicates that alpha1 and beta1 integrin expression coincides temporally with the intracellular proliferation and reorientation of myofilaments. The similarity in distribution patterns of alpha1 and beta1 integrins during development suggests that the two integrin subunits are synchronously expressed during development and do not appear sequentially. (J Histochem Cytochem 46:119-125, 1998) PMID- 9405502 TI - Immunohistochemical detection of the MUC1 gene product in human cancers grown in scid mice. AB - Alterations in mucin expression have been detected in many clinically relevant cancers and, in particular, the polymorphic epithelial mucin, encoded by the MUC1 gene, has attracted considerable attention. We investigated its expression in human breast, colon, ovarian, lung, and skin cancer cells and their metastases grown in severe combined immunodeficient (scid) mice using three different monoclonal antibodies (HMFG-1, HMFG-2, and SM3). Four of five breast cancer cell lines, three of five colon cancer cell lines, two of three small-cell carcinoma of the lung cell lines, and A 431 cells all expressed the MUC1 gene product. Neuraminidase predigestion often enhanced HMFG-1 immunoreactivity, which was more widespread and stronger than SM3 immunoreactivity. A considerable heterogeneity of MUC1 gene product expression was observed in the same tumors grown in different mice. The binding pattern between single-cell/small-cell clusters (up to 10 cells) and larger cell number aggregates varied. The results indicate that the MUC1 gene expression both in primary tumors and metastases is not tightly controlled within a particular tumor cell line. Because of this heterogeneous antigen expression in vivo, it appears impossible to target all metastatic deposits by a single monoclonal antibody directed against the MUC1 gene product. (J Histochem Cytochem 46:127-134, 1998) PMID- 9405503 TI - Functional role of peptidergic anterior lobe neurons in male sexual behavior of the snail Lymnaea stagnalis. AB - A morphologically defined group of peptidergic neurons in the CNS of the hermaphroditic snail, Lymnaea stagnalis, is concerned with the control of a very specific element of male sexual behavior. These neurons are located in the anterior lobe of the right cerebral ganglion (rAL). By using chronically implanted electrodes, we show that the rAL neurons are selectively active during eversion of the penis-carrying structure, the preputium. The preputium is normally contained inside the body cavity and is everted during copulation in the male role. Electrical stimulation of the rAL neurons through the implanted electrodes, induced eversion of the preputium in vivo. Injection of APGWamide (Ala-Pro-Gly-Try-NH2), a small neuropeptide that is present in all rAL neurons, induced eversion of the preputium. Application of APGWamide to in vitro preparations of the preputium caused relaxation of this organ. In contrast, injection of the neuropeptide conopressin, which is co-localized with APGWamide in 60% of the rAL neurons, did not induce any behavior associated with male sexual activities. These results show that the neurons of the rAL can induce an eversion of the preputium as occurs during male copulation by release of APGWamide during a period of electrical activity. PMID- 9405504 TI - Mechanisms of within- and cross-modality suppression in the superior colliculus. AB - The present studies were initiated to explore the basis for the response suppression that occurs in cat superior colliculus (SC) neurons when two spatially disparate stimuli are presented simultaneously or in close temporal proximity to one another. Of specific interest was examining the possibility that suppressive regions border the receptive fields (RFs) of unimodal and multisensory SC neurons and, when activated, degrade the neuron's responses to excitatory stimuli. Both within- and cross-modality effects were examined. An example of the former is when a response to a visual stimulus within its RF is suppressed by a second visual stimulus outside the RF. An example of the latter is when the response to a visual stimulus within the visual RF is suppressed when a stimulus from a different modality (e. g., auditory) is presented outside its (i.e., auditory) RF. Suppressive regions were found bordering visual, auditory, and somatosensory RFs. Despite significant modality-specific differences in the incidence and effectiveness of these regions, they were generally quite potent regardless of the modality. In the vast majority (85%) of cases, responses to the excitatory stimulus were degraded by >/=50% by simultaneously stimulating the suppressive region. Contrary to expectations and previous speculations, the effects of activating these suppressive regions often were quite specific. Thus powerful within-modality suppression could be demonstrated in many multisensory neurons in which cross-modality suppression could not be generated. However, the converse was not true. If an extra-RF stimulus inhibited center responses to stimuli of a different modality, it also would suppress center responses to stimuli of its own modality. Thus when cross-modality suppression was demonstrated, it was always accompanied by within-modality suppression. These observations suggest that separate mechanisms underlie within- and cross-modality suppression in the SC. Because some modality-specific tectopetal structures contain neurons with suppressive regions bordering their RFs, the within-modality suppression observed in the SC simply may reflect interactions taking place at the level of one input channel. However, the presence of modality-specific suppression at the level of one input channel would have no effect on the excitation initiated via another input channel. Given the modality-specificity of tectopetal inputs, it appears that cross-modality interactions require the convergence of two or more modality-specific inputs onto the same SC neuron and that the expression of these interactions depends on the internal circuitry of the SC. This allows a cross-modality suppressive signal to be nonspecific and to degrade any and all of the neuron's excitatory inputs. PMID- 9405505 TI - Intrinsic membrane characteristics distinguish two subsets of nociceptive modulatory neurons in rat RVM. AB - Pain modulating neurons of the rostral ventromedial medulla (RVM) include three physiologically distinct classes of neurons in intact, anesthetized animals: and cells that change their activity before the onset of withdrawal reflexes and cells, which have activity unrelated to withdrawal reflexes. A previous in vitro intracellular study demonstrated that the RVM contains two types of neurons that are distinguished by their action-potential characteristics. The present in vivo intracellular study examined whether these intracellularly recorded action potential characteristics are correlated with the physiological response properties of RVM neurons recorded. RVM neurons exhibited two distinct types of action potentials in vivo. Fast-spike (FS) neurons (n = 30) had short-duration action potentials (0.27 +/- 0.02 (SE) ms at half amplitude) and biphasic afterhyperpolarizations with a characteristic rapid overshooting spike repolarization. Slow-spike (SS) neurons (n = 25) had longer duration action potentials (0.44 +/- 0.02 ms at half-amplitude) due to a slower-spike repolarization rate and monophasic afterhyperpolarization. and cell classes included both FS and SS neurons. FS and neurons had an early onset response to noxious heat stimulation. SS and cells showed a delayed onset response to noxious heat. cells (n = 13) were all SS cells. Among the SS neurons, only cells had action potentials longer than 0. 45 ms (n = 9). FS and SS neurons were intermingled throughout the RVM. The majority of intracellularly labeled cells (n = 15) had fusiform somata with two to five fine caliber primary dendrites and a predominantly mediolateral orientation of the long axis of their dendritic tree. All labeled FS cells (n = 5) had large, multipolar somata with four to nine large caliber primary dendrites. The present study defines in vivo membrane and morphological characteristics of RVM neurons that correlate with physiological differences and may be used for identification of nociceptive modulatory RVM neurons in slice preparations. PMID- 9405506 TI - Role of neuropeptides encoded on CDCH-1 gene in the organization of egg-laying behavior in the pond snail, Lymnaea stagnalis. AB - Egg laying in the pond snail Lymnaea stagnalis is triggered by a discharge of the neuroendocrine caudodorsal cells (CDCs). The CDCs expresses three different caudorsal cell hormone (CDCH) genes. This gene family expresses, in total, 11 different peptides among which is the ovulation hormone. Besides the CDCs, the CDCH gene family is expressed in other central and peripheral neurons. In this study, we investigated the roles the different CDCH peptides play in the organization of egg-laying behavior. Egg-laying behavior is a sequence of stereotyped movements in which three phases can be distinguished: resting, turning, and oviposition. We have used the excitation of right pedal N (RPeN) motor neurons as a simple analogue of shell-turning behavior, one of the elements of egg-laying behavior. RPeN motor neurons were inhibited during the resting phase of egg laying but were subsequently excited at the onset of and during the turning phase. The excitatory effect could be evoked by application of beta3-CDCP on RPeN motor neurons in the CNS as well as in isolation but not by the ovulation hormone, alpha-CDCP or Calfluxin, the other CDCH-1 peptides tested. The ovulation hormone itself caused inhibition of RPeN motor neurons. Anti-CDCH-1 positive fiber tracts were found close to the cell bodies and axons of the RPeN motor neurons. Electrical stimulation of a nerve that contains these fibers resulted in excitation of the RPeN motor neurons. The effects of injection of CDCH-1 peptides into intact animals correlated well with the effects of these peptides on RPeN motor neurons. Injection of beta3-CDCP or alpha-CDCP into intact animals resulted in immediate turning behavior in the absence of egg laying itself. The ovulation hormone and Calfluxin had no immediate effect on the behavior. Furthermore, our data indicate that the individual CDCH-1 peptides act on different targets. PMID- 9405507 TI - Estimating the distribution of synaptic reliabilities. AB - Using whole cell recording from CA1 hippocampal pyramidal neurons in slices, we examined the progressive decrease of N-methyl-D-aspartate receptor-mediated synaptic responses in the presence of the open-channel blocker MK-801. Previous studies analyzing this decrease have proposed that hippocampal synapses fall into two distinct classes of release probabilities, whereas studies based on other methods indicate a broad distribution of synaptic reliabilities exists. Here we derive the theoretical relationship between the MK-801-mediated decrease in excitatory postsynaptic current amplitudes and the underlying distribution of synaptic reliabilities. We find that the MK-801 data are consistent with a continuous distribution of synaptic reliabilities, in agreement with studies examining individual synapses. In addition, changes in the MK-801-mediated decrease in response size as a consequence of altering release probability are consistent with this continuous distribution of synaptic reliabilities. PMID- 9405508 TI - Encoding of object curvature by tactile afferents from human fingers. AB - Isolated responses were recorded from fibers in the median nerves of human subjects by using microneurography. Mechanoreceptive afferent fibers with receptive fields on the fingerpads were selected. The fingers were immobilized and spherical stimuli were applied passively to the receptive field with a contact force of 40-, 60-, or 80-g weight. The radii of the spheres were 1.92, 2.94, 5.81, or 12.4 mm or infinity (flat); the corresponding curvatures, given by the reciprocal of the radii, were 694, 340, 172, 80.6, or 0 m-1, respectively. When the spheres were applied to the receptive field center of slowly adapting type I afferents (SAIs), the response increased as the curvature of the sphere increased and also increased as the contact force increased. All SAIs behaved in the same way except for a scaling factor proportional to the sensitivity of the afferent. When a sphere was located at different positions in the receptive field, the shape of the resulting response profile reflected the shape of the sphere; for more curved spheres the profile was higher and narrower (increased peak and decreased width). Slowly adapting type II afferents (SAIIs) showed different response characteristics from the SAIs when spheres were applied to their receptive field centers. As the curvature of the stimulus increased from 80.6 to 172 m-1, the response increased. However, further increases in curvature did not result in further increases in response. An increase in contact force resulted in an increase in the response of SAIIs; this increase was proportionately greater than it was for SAIs. For SAIIs, the shape of the receptive field profile did not change when the curvature of the stimulus changed. For fast-adapting type I afferents (FAIs), the responses were small and did not change systematically with changes in curvature or contact force. Fast adapting type II afferents (FAIIs) did not respond to our stimuli. Human SAIs, FAIs, and FAIIs behaved like monkey SAIs, FAIs, and FAIIs, respectively. The response of the SAI population contains accurate information about the shape of the sphere and its position of contact on the finger and also indicates contact force. Conversely, whereas SAIIs possess a greater capacity to encode changes in contact force, they provide only coarse information on local shape. PMID- 9405509 TI - Perceptual learning of spatial localization: specificity for orientation, position, and context. AB - Discrimination of simple visual attributes can improve significantly with practice. We have trained human observers to perform peripherally presented tasks involving the localization of short line segments and examined the specificity of the learning for the visual location, orientation, and geometric arrangement of the trained stimulus. Several weeks of training resulted in dramatic threshold reductions. The learning was specific for the orientation and location of the trained stimulus, indicating the involvement of the earliest cortical stages in the visual pathway where the orientation and location of stimuli are mapped with highest resolution. Furthermore, improvement was also specific for both the configuration of the trained stimulus and the attribute of the stimulus that was under scrutiny during training. This degree of specificity suggests that the learning cannot be achieved by cortical recruitment alone, as proposed in current models, but is likely to involve a refinement of lateral interactions within the cortex and possibly a gating of lower level changes by attentional mechanisms. PMID- 9405510 TI - Functional development of intrinsic properties in ganglion cells of the mammalian retina. AB - Senosory neurons manifest pronounced changes in excitability during maturation, but the factors contributing to this ubiquitous developmental phenomenon are not well understood. To assess the contribution of intrinsic membrane properties to such changes in excitability, in the present study whole cell patch-clamp recordings were made from developing ganglion cells in the intact retina of postnatal rats. During a relatively brief developmental period (postnatal days P7 P27) ganglion cells exhibited pronounced changes in the discharge patterns generated by depolarizing current injections. The youngest cells (P7-P17) typically responded to maintained depolarizations with only a single spike or a rapidly adapting discharge pattern. In contrast, the predominant response mode of more mature cells (P21-P27) was a series of repetitive discharges that lasted for the duration of the depolarization period, and by P25 all cells responded in this manner. These functional changes characterized all three morphologically defined cell classes identified by intracellular labeling with Lucifer yellow. To determine if expression of the potassium current (Ia) and the kinetics of the Na channel related to the increased excitability of developing ganglion cells described above, current- and voltage-clamp recordings were made from individual neurons. The different firing patterns manifested by developing retinal ganglion cells did not reflect the presence or absence of the Ia conductance, although cells expressing Ia tended to generate spikes of shorter duration. With maturation the speed of recovery from inactivation of the Na current increased markedly and this related to the increased excitability of developing ganglion cells. Neurons yielding only a single spike to maintained depolarization were characterized by the slowest speed of recovery; cells with rapidly adapting discharges showed a faster recovery and those capable of repetitive firing recovered fastest from Na-channel inactivation. It is suggested that these changes in intrinsic membrane properties may relate to the different functional roles subserved by ganglion cells during development. PMID- 9405511 TI - Responses of MT and MST neurons to one and two moving objects in the receptive field. AB - To test the effects of complex visual motion stimuli on the responses of single neurons in the middle temporal visual area (MT) and the medial superior temporal area (MST) of the macaque monkey, we compared the response elicited by one object in motion through the receptive field with the response of two simultaneously presented objects moving in different directions through the receptive field. There was an increased response to a stimulus moving in a direction other than the best direction when it was paired with a stimulus moving in the best direction. This increase was significant for all directions of motion of the non best stimulus and the magnitude of the difference increased as the difference in the directions of the two stimuli increased. Similarly, there was a decreased response to a stimulus moving in a non-null direction when it was paired with a stimulus moving in the null direction. This decreased response in MT did not reach significance unless the second stimulus added to the null direction moved in the best direction, whereas in MST the decrease was significant when the second stimulus direction differed from the null by 90 degrees or more. Further analysis showed that the two-object responses were better predicted by taking the averaged response of the neuron to the two single-object stimuli than by summation, multiplication, or vector addition of the responses to each of the two single-object stimuli. Neurons in MST showed larger modulations than did neurons in MT with stimuli moving in both the best direction and in the null direction and the average better predicted the two-object response in area MST than in area MT. This indicates that areas MT and MST probably use a similar integrative mechanisms to create their responses to complex moving visual stimuli, but that this mechanism is further refined in MST. These experiments show that neurons in both MT and MST integrate the motion of all directions in their responses to complex moving stimuli. These results with the motion of objects were in sound agreement with those previously reported with the use of random dot patterns for the study of transparent motion in MT and suggest that these neurons use similar computational mechanisms in the processing of object and global motion stimuli. PMID- 9405512 TI - Analysis of AMPA receptor properties during postnatal development of mouse hippocampal astrocytes. AB - Glial cells in the mammalian brain express various types of voltage- and ligand gated ion channels, including glutamate receptors (GluRs) of the alpha-amino-3 hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-subtype. In the present study we followed developmental changes in the functional properties of AMPA receptor (AMPA-R) channels expressed by astrocytes of the mouse hippocampus between postnatal days (P) 5-35 to learn more about the physiological significance of these glial receptors. A fast concentration clamp technique was applied to cells acutely isolated from the CA1 stratum radiatum subregion to quantitatively analyze rapidly activating and desensitizing receptor responses. The equilibrium responses of glutamate and kainate differed between P5 and P12. Although the maximum current induced by kainate was almost the same at all developmental stages, a steep rise in the maximum glutamate response was observed within the same time range. Between P5 and P12 there was an increase in the potentiation of AMPA-R currents with cyclothiazide (CTZ); at the same time, the dissociation kinetics of CTZ became significantly slower. These changes in the pharmacological properties suggested a variation in splice variant expression. With proceeding maturation, we observed an increase in the degree of desensitization of the glutamate- and AMPA-induced receptor currents. In addition to the shift in flip/flop splicing, these findings could hint at a developmental regulation of RNA editing in the arginine/glycine site. Altogether, the present results demonstrate changes in astrocytic AMPA-R functioning early in postnatal development, although after P12 the receptor properties remained almost constant. Although the overall Ca2+ permeability did not vary during development, the prolonged receptor opening in the early postnatal period causes an enhanced Na+/Ca2+ influx into the immature astrocytes. This could influence glial proliferation and differentiation during CNS ontogenesis. PMID- 9405513 TI - Reorganization of the raccoon cuneate nucleus after peripheral denervation. AB - The effects of peripheral nerve transection on the cuneate nucleus were studied in anesthetized raccoons using extracellular, single-unit recordings. The somatotopic organization of the cuneate nucleus first was examined in intact, control animals. The cuneate nucleus in the raccoon is organized with the digits represented in separate cell clusters. The dorsal cap region of the cuneate nucleus contains a representation of the claws and hairy skin of the digits. Within the representation of the glabrous skin, neurons with rapidly adapting properties tended to be segregated from those with slowly adapting properties. The representations of the distal and proximal pads on a digit also were segregated. Electrical stimulation of two adjacent digits provided a detailed description of the responses originating from the digit that contains the tactile receptive field (the on-focus digit) and from the adjacent (off-focus) digit. Stimulation of the on-focus digit produced a short latency excitation in all 99 neurons tested, with a mean of 10.5 ms. These responses had a low threshold (426 microA). Stimulation of an off-focus digit activated 65% of these neurons. These responses had a significantly longer latency (15.3 ms) than on-focus responses and the threshold was more than twice as large. Two to five months after amputation of digit 4, 97 cells were tested with stimulation of digits 3 and 5. A total of 44 were in the intact regions of the cuneate nucleus. They had small receptive fields on intact digits and their responses to electrical stimulation did not differ from the control neurons. The remaining 53 neurons were judged to be deafferented and in the fourth digit region on the basis of their location with respect to intact neurons. All but two of these cells had receptive fields that were much larger than normal, often including more than one digit and part of the palm. When compared with the off-focus control neurons, their responses to electrical stimulation had lower thresholds and an increased response probability and magnitude. The latencies of these cells did not decrease, however, and were the same as the off-focus control values. The enhanced responses of the deafferented neurons to adjacent digit stimulation indicate that there is a strengthening of synapses that were previously ineffective. The increased proportion of neurons that could be activated after amputation suggests that there is also a growth of new connections. This experiment demonstrates that reorganization in the adult somatotopic system does occur at the level of the dorsal column nuclei. As a consequence, many of the changes reported at the cortex and thalamus may be due to the changes occurring at this first synapse in the somatosensory pathway. PMID- 9405515 TI - Neural circuit mediating tentacle withdrawal in Helix aspersa, with specific reference to the competence of the motor neuron C3. AB - The tentacle withdrawal reflex in the terrestrial snail Helix aspersa involves bending and retraction of the tentacles. When elicited by mechanical stimulation of the tentacle, the reflex is mediated by the conjoint action of the central and peripheral nervous systems. The neural circuit underlying the stimulus-response pathways was studied in vitro using a combination of morphological and physiological techniques. Sensory input caused by stimulation of the nose (situated at the superior tentacle's tip) first passes into the tentacle ganglion. Motor fibers are likely excited in the tentacle ganglion to form a peripheral stimulus-response pathway. While still in the tentacle ganglion, the excitation caused by a brief stimulus is transformed into a prolonged neuronal discharge. This modified signal travels, via the olfactory nerve, to the cerebral ganglion where it excites the giant motor neuron C3 along with numerous smaller motor neurons. Afferent input to C3 also arrives from several other sources. The afferent convergence is followed by a marked divergence of C3's output. C3 innervates the muscles mediating both tentacle retraction and tentacle bending through multiple cerebral nerves. Thus C3's pattern of effector innervation allows this single cell to elicit and coordinate both components of the tentacle withdrawal reflex. Lesion experiments indicate that C3 is responsible for 85% of the central contribution to tentacle retraction, though C3 is actually sufficient to mediate maximal muscle contraction as evidenced by intracellular stimulation. In addition to C3, three groups of putative central motor neurons were identified through nerve backfills and nerve recordings. The additional motor neurons mediating tentacle retraction are important for maximizing the rate of muscle contraction, whereas those mediating tentacle bending are likely more important for nondefensive behaviors. These neurons are arranged in parallel with C3, but unlike C3, each of these neurons innervates only a single effector or portion thereof. Given C3's direct innervation of multiple effectors and its sufficiency to evoke strong responses in those effectors, we conclude that C3 is paramount in eliciting and coordinating tentacle withdrawal. PMID- 9405514 TI - Characterization of gating and peptide block of mSlo, a cloned calcium-dependent potassium channel. AB - The 20 amino acid Shaker inactivation peptide blocks mSlo, a cloned calcium dependent potassium channel. Changing the charge and degree of hydrophobicity of the peptide alters its blocking kinetics. A "triple mutant" mSlo channel was constructed in which three amino acids (T256, S259, and L262), equivalent to those identified as part of the peptide's receptor site in the S4-S5 cytoplasmic loop region of the Shaker channel, were mutated simultaneously to alanines. These mutations produce only limited changes in the channel's susceptibility to block by a series of peptides of varying charge and hydrophobicity but do alter channel gating. The triple mutant channel shows a significant shift in its calcium activation curve as compared with the wild-type channel. Analysis of the corresponding single amino acid mutations shows that mutation at position L262 causes the most dramatic change in mSlo gating. These results suggest that the three amino acids mutated in the mSlo S4-S5 loop may contribute to, but are not essential for, peptide binding. On the other hand, they do play a critical role in the channel's calcium-sensing mechanism. PMID- 9405516 TI - Two types of ACh receptors contribute to fast channel gating on mouse skeletal muscle. AB - Single-channel recordings from mouse C2 myotubes indicate that maturation of skeletal muscle is accompanied by the appearance of two types of fast acetylcholine (ACh) receptor channels that are each functionally distinct from the embryonic receptor type present at early stages of differentiation. The embryonic receptor type has a low conductance (45 pS) and long channel open time, rendering slowly decaying synaptic currents. One fast channel type that appears during muscle maturation is distinguished from the embryonic receptor type on the basis of both higher conductance (65 pS) and shorter open time. However, single channel recordings from differentiated mouse skeletal muscle cell line (C2) point to the existence of a second fast receptor type, which has a conductance similar to the embryonic receptor type (45 pS), yet significantly reduced mean channel open time. Analyses of individual channel function at high ACh concentrations directly demonstrate the coexistence of two kinetically distinct types of 45 pS ACh receptors. Openings by fast type and slow embryonic type of 45 pS receptors occurred in bursts, allowing distinction on the basis of both mean open time and open probability for individual receptors. The embryonic type of 45 pS receptor has an open time approximately twofold longer than the fast-receptor counterpart. Additional differences were reflected in the open probability distributions for fast and slow 45 pS receptor types. Both types of 45 pS receptor were kinetically distinguishable from the 65 pS receptor. We found no support for the idea that the slow and fast 45 pS receptor types result from the interconversion of dual gating modes involving the same receptor protein. Our results are consistent with the idea that the acquisition of fast synaptic current decay, required at mature neuromuscular synapses, is the result of the up-regulation of two distinct fast types of nicotinic ACh receptors during skeletal muscle development. PMID- 9405517 TI - Stretch of quadriceps inhibits the soleus H reflex during locomotion in decerebrate cats. AB - Previously, it has been demonstrated that afferent signals from the quadriceps muscles can suppress H reflexes in humans during passive movements of the leg. To establish whether afferent input from quadriceps contributes to the modulation of the soleus H reflex during locomotion, the soleus H reflex was conditioned with stretches of the quadriceps muscle during bouts of spontaneous treadmill locomotion in decerebrate cats. We hypothesized that 1) in the absence of locomotion such conditioning would lead to suppression of the soleus H reflex and 2) this would be retained during periods of locomotor activity. In the absence of locomotion, slow sinusoidal stretches (0.2 Hz, 8 mm) of quadriceps cyclically modulated the amplitude of the soleus H reflex. The H reflex amplitude was least during the lengthening of the quadriceps and greatest as quadriceps shortened. Further, low-amplitude vibrations (48-78 micron) applied to the patellar tendon suppressed the reflex, indicating that the muscle spindle primaries were the receptor eliciting the effect. During bouts of locomotion, ramp stretches of quadriceps were applied during the extensor phase of the locomotor rhythm. Soleus H reflexes sampled at two points during the stance phase were reduced compared with phase-matched controls. The background level of the soleus electromyographic activity was not influenced by the applied stretches to quadriceps, either during locomotion or in the absence of locomotion. This indicates that the excitability of the soleus motoneuron pool was not influenced by the stretching of quadriceps, and that the inhibition of the soleus H reflex is due to presynaptic inhibition. We conclude that group Ia afferent feedback from quadriceps contributes to the regulation of the soleus H reflex during the stance phase of locomotion in decerebrate cats. This afferent mediated source of regulation of the H reflex, or monosynaptic stretch reflex, would allow for rapid alterations in reflex gain according to the dynamic needs of the animal. During early stance, this source of regulation might suppress the soleus stretch reflex to allow adequate yielding at the ankle and facilitate the movement of the body over the foot. PMID- 9405518 TI - Directional control of planar human arm movement. AB - We examined the patterns of joint kinematics and torques in two kinds of sagittal plane reaching movements. One consisted of movements from a fixed initial position with the arm partially outstretched, to different targets, equidistant from the initial position and located according to the hours of a clock. The other series added movements from different initial positions and directions and >40-80 cm distances. Dynamic muscle torque was calculated by inverse dynamic equations with the gravitational components removed. In making movements in almost every direction, the dynamic components of the muscle torques at both the elbow and shoulder were related almost linearly to each other. Both were similarly shaped, biphasic, almost synchronous and symmetrical pulses. These findings are consistent with our previously reported observations, which we termed a linear synergy. The relative scaling of the two joint torques changes continuously and regularly with movement direction. This was confirmed by calculating a vector defined by the dynamic components of the shoulder and elbow torques. The vector rotates smoothly about an ellipse in intrinsic, joint torque space as the direction of hand motion rotates about a circle in extrinsic Cartesian space. This confirms a second implication of linear synergy that the scaling constant between the linearly related joint torques is directionally dependent. Multiple linear regression showed that the torque at each joint scales as a simple linear function of the angular displacement at both joints, in spite of the complex nonlinear dynamics of multijoint movement. The coefficients of this function are independent of the initial arm position and movement distance and are the same for all subjects. This is an unanticipated finding. We discuss these observations in terms of the hypothesis that voluntary, multiple degrees of freedom, rapid reaching movements may use rule-based, feed-forward control of dynamic joint torque. Rule-based control of joint torque with separate dynamic and static controllers is an alternative to models such as those based on the equilibrium point hypotheses that rely on a positionally based controller to produce both dynamic and static torque components. It is also an alternative to feed-forward models that directly solve the problems of inverse dynamics. Our experimental findings are not necessarily incompatible with any of the alternative models, but they describe new, additional findings for which we need to account. The rules are chosen by the nervous system according to features of the kinematic task to couple muscle contraction at the shoulder and elbow in a linear synergy. Speed and load control preserves the relative magnitudes of the dynamic torques while directional control is accomplished by modulating them in a differential manner. This control system operates in parallel with a positional control system that solves the problems of postural stability. PMID- 9405519 TI - Kinetics of muscarinic reduction of IsAHP in hippocampal neurons: effects of acetylcholinesterase inhibitors. AB - The present experiments were designed to elucidate the time frame in which an evoked cholinergic impulse decreases the Ca2+-dependent K+ current (IsAHP) in hippocampal CA1 neurons, and to determine to what extent acetylcholinesterase (AChE) inhibitors enhance the efficacy of the cholinergic impulse. Whole cell voltage-clamp recordings were performed on hippocampal CA1 neurons of rat brain slices and IsAHPs were evoked by constant depolarizing pulses. Cholinergic afferent fibers in stratum oriens were stimulated electrically and the time interval between the afferent stimulus and the depolarizing pulse was varied from 1 to 30 s. In slices perfused with the standard external medium, the afferent stimulus caused a profound decrease in the following IsAHP only when the stimulus preceded the depolarizing pulse by 1-2 s. The stimulus was without effects on the IsAHP when applied >/=5s before the depolarizing pulse. The effects of the afferent stimulus were greatly enhanced in CA1 neurons exposed to the catalytic AChE inhibitors neostigmine, physostigmine, or 9-amino-1,2,3, 4-tetrahydro acridine. A substantial decrease in the IsAHP was observed even when the stimulus preceded the depolarizing pulse by >/=30 s. However applications of peripheral site AChE inhibitors decamethonium and propidium caused only minor or no enhancement of the IsAHP reduction after the afferent stimulus. We suggest in physiological conditions that muscarinic modulation of ionic conductances of CNS neurons has a limited time course after a cholinergic impulse and that the modulation is greatly enhanced and prolonged when catalytic activities of AChEs are suppressed pharmacologically. PMID- 9405520 TI - Somatostatin depresses excitatory but not inhibitory neurotransmission in rat CA1 hippocampus. AB - In rat CA1 hippocampal pyramidal neurons (HPNs), somatostatin (SST) has inhibitory postsynaptic actions, including hyperpolarization of the membrane at rest and augmentation of the K+ M-current. However, the effects of SST on synaptic transmission in this brain region have not been well-characterized. Therefore we used intracellular voltage-clamp recordings in rat hippocampal slices to assess the effects of SST on pharmacologically isolated synaptic currents in HPNs. SST depressed both (R, S)-alpha-amino-3-hydroxy-5 methylisoxazole-4-propionic acid (AMPA)/kainate and N-methyl--aspartate (NMDA) receptor-mediated excitatory postsynaptic currents (EPSCs) in a reversible manner, with an apparent IC50 of 22 nM and a maximal effect at 100 nM. In contrast, SST at concentrations up to 5 microM had no direct effects on either gamma-aminobutyric acid-A (GABAA) or GABAB receptor-mediated inhibitory postsynaptic currents (IPSCs). The depression of EPSCs by SST was especially robust during hyperexcited states when polysynaptic EPSCs were present, suggesting that this peptide could play a compensatory role during seizurelike activity. SST effects were greatly attenuated by the alkylating agent N ethylmaleimide, thus implicating a transduction mechanism involving the Gi/Go family of G-proteins. Use of 2 M Cs+ in the recording electrode blocked the postsynaptic modulation of K+ currents by SST, but did not alter the effects of SST on EPSCs, indicating that postsynaptic K+ currents are not involved in this action of SST. However, 2 mM external Ba2+ blocked the effect of SST on EPSCs, suggesting that presynaptic K+ channels or other presynaptic mechanisms may be involved. These findings and previous results from our laboratory show that SST has multiple inhibitory effects in hippocampus. PMID- 9405521 TI - Heterogeneous voltage dependence of inward rectifier currents in spiral ganglion neurons. AB - Inward rectification was characterized in neonatal spiral ganglion neurons maintained in tissue culture. Whole cell current and voltage-clamp techniques were used to show that the hyperpolarization-activated cationic (Ih) current underlies most or all of the inward rectification demonstrated in these neurons. The average reversal potential (-41.3 mV) and cesium sensitivity were typical of that found in other neurons and cell types. What was unique about the hyperpolarization-activated currents, however, was that the half-maximal voltages (V1/2) and slope factors (k) that characterized Ih current activation were graded from neuron to neuron. Voltage-clamp recordings made with standard bath and pipette solutions revealed V1/2 values that ranged from -78.1 to -122.1 mV, with slope factors from 7.6 to 13.1. These gradations in the voltage-dependent features of the Ih current did not result from variability in the recording conditions because independently measured Na+ current-to-voltage relationships were found to be uniform (peak current at -20 mV). Moreover, the range and average V1/2 and slope values could be altered with activators [8-(4 chlorophenylthio) adenosine 3',5'-cyclic monophosphate in combination with okadaic acid] or inhibitors {N-[2-(p-bromocinnamylamino)ethyl]-5 isoquinolinesulfonamide}of protein indicating that Ih current heterogeneity most likely resulted from differential phosphorylation. PMID- 9405522 TI - Metabotropic glutamate receptors regulate N-methyl-D-aspartate-mediated synaptic transmission in nucleus accumbens. AB - We recorded intracellularly from core nucleus accumbens (NAcc) neurons in brain slices to study the regulation by metabotropic glutamate receptors (mGluRs) of pharmacologically isolated N-methyl--aspartate-mediated excitatory postsynaptic currents (NMDA-EPSCs). Monosynaptic NMDA-EPSCs, evoked by local stimulation, were isolated by superfusion of the non-NMDA and gamma-aminobutyric acid-A (GABAA) receptor antagonists, 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 10 microM) and bicuculline (15 microM), respectively. Trans-1-aminocyclopentane-1,3-decarboxylic acid (trans-ACPD; 50 microM), a nonspecific group 1 and 2 mGluR agonist, had no effect on resting membrane potential (RMP) or input resistance of NAcc neurons. However, it consistently decreased NMDA-EPSC areas (time integrals) dose dependently (1-100 microM; EC50 = 8 microM) and reversibly. The specific group 1 mGluR agonists quisqualate (1-4 microM) and (RS)-3, 5-dihydroxyphenylglycine (DHPG; 100 microM) did not mimic the trans-ACPD effect on NMDA-EPSCs, nor did exposure of the slice to the group 1 mGluR antagonist (+)-2-amino-3 phosphonopropionic acid (-AP3, 0.4 mM) inhibit the trans-ACPD effect. The putative mGluR1 and mGluR2 antagonist (+)-alpha-methyl-4-carboxyphenylglycine (MCPG) at 0.5 mM failed to antagonize trans-ACPD effects but at 1 mM blocked them. Both the group 2 mGluR agonist (2S,3S, 4S)-alpha-(carboxycyclopropyl) glycine (-CCG-I, 2 microM) and the group 3 mGluR specific agonist (+)-2-amino-4 phosphonobutyric acid (-AP4, 20 microM) attenuated NMDA-EPSC areas; the effect of -AP4 was blocked by the group 3 antagonist (S)-2-amino-2-methyl-4 phosphonobutanoic acid (MAP4; 0.5 mM). Exogenously applied NMDA, in the presence of tetrodotoxin to prevent presynaptic effects, induced inward currents that were decreased by 20 microM -AP4 but not by 10 microM trans-ACPD. These findings suggest that NMDA receptor-mediated neurotransmission in NAcc is under dual inhibitory regulation by group 2 and 3 metabotropic receptor subtypes: -AP4 sensitive receptors located postsynaptically and those sensitive to trans-ACPD located presynaptically. PMID- 9405523 TI - Metabotropic glutamate receptors are involved in long-term potentiation in isolated slices of rat medial frontal cortex. AB - The prelimbic region of medial frontal cortex in the rat receives a direct input from the hippocampus and this functional connection is essential for aspects of spatial memory. Activity-dependent changes in the effectiveness of synaptic transmission in the medial frontal cortex, namely long-term potentiation (LTP) and long-term depression (LTD) can persist for tens of minutes or hours and may be the basis of learning and memory storage. Glutamatergic activation of ionotropic receptors is required to induce both LTP and LTD. We now present evidence of the involvement of metabotropic glutamate receptors in LTP in isolated slices of frontal cortex. Repetitive bursts of stimulation at theta frequencies (TBS) were applied to layer II, and monosynaptic EPSPs were monitored in layer V neurons of the prelimbic area. TBS was found to be more effective at inducing LTP than tetanic stimulation at 100 Hz and produced LTP that lasted >30 min in 8 out of 14 neurons. Tetanic stimulation at 100 Hz in the presence of the N-methyl--aspartate (NMDA)-antagonist 2-amino-5-phosphonopentanoate (AP5) was reported to be a reliable method of inducing LTD in prelimbic cortex (). However we found that this protocol did not facilitate the induction of LTD. The role of metabotropic glutamate receptors (mGluR) in LTP was assessed by using the selective, broad-spectrum antagonist (R, S)-alpha-methyl-4- carboxyphenylglycine (MCPG). This drug significantly reduced the incidence of LTP after TBS to only 1 of 14 neurons (P < 0.02, chi2 test). The pooled responses to TBS in MCPG showed significantly reduced potentiation [(P < 0.02, analysis of variance (ANOVA)]. The broad-spectrum mGluR agonist (1S, 3R)-1-aminocyclopentane-1,3-dicarboxylic acid (ACPD) and the selective group I agonist S-3 hydroxyphenylglycine(S-3HPG) both produced membrane depolarization, an increase in number of spikes evoked by depolarizing current pulses, and a reduction in the afterhyperpolarization. Similar effects were produced by these agonists even when synaptic transmission was blocked by use of the gamma-aminobutyric acid-B (GABAB) receptor agonist, 200 microM baclofen, which suggests that group I mGluRs are present on layer V neurons. We conclude that mGluRs participate in the production of LTP in prelimbic cortex, and that this excitatory effect could be mediated by the postsynaptic group I mGluRs. PMID- 9405524 TI - Motor patterns and kinematics during backward walking in the pacific giant salamander: evidence for novel motor output. AB - Kinematic and motor patterns during forward and backward walking in the salamander Dicamptodon tenebrosus were compared to determine whether the differences seen in mammals also apply to a lower vertebrate with sprawling posture and to measure the flexibility of motor output by tetrapod central pattern generators. During treadmill locomotion, electromyograms (EMGs) were recorded from hindlimb muscles of Dicamptodon while simultaneous high-speed video records documented movement of the body, thigh, and crus and allowed EMGs to be synchronized to limb movements. In forward locomotion, the trunk was lifted above the treadmill surface. The pelvic girdle and trunk underwent smooth side-to-side oscillations throughout the stride. At the beginning of the stance phase, the femur was protracted and the knee joint extended. The knee joint initially flexed in early stance and then extended as the foot pushed off in late stance, reaching maximum extension just before foot lift-off. The femur retracted steadily throughout the stance. In the swing phase, the femur rapidly protracted, and the leg was brought forward in an "overhand crawl" motion. In backward walking, the body frequently remained in contact with the treadmill surface. The pelvic girdle, trunk, and femur remained relatively still during stance phase, and most motion occurred at the knee joint. The knee joint extended throughout most of stance, as the body moved back, away from the stationary foot. The knee flexed during swing. Four of five angles showed significantly smaller ranges in backward than in forward walking. EMGs of forward walking showed that ventral muscles were coactive, beginning activity just before foot touchdown and ceasing during the middle of stance phase. Dorsal muscles were active primarily during swing. Backward locomotion showed a different pattern; all muscles except one showed primary activity during the swing phase. This pattern of muscle synergy in backward walking never was seen in forward locomotion. Also, several muscles demonstrated lower burst rectified integrated areas (RIA) or durations during backward locomotion. Multivariate statistical analysis of EMG onset and RIA completely separated forward and backward walking along the first principal component, based on higher RIAs, longer durations of muscle activity, and greater synergy between ventral muscles during early stance in forward walking. Backward walking in Dicamptodon uses a novel motor pattern not seen during forward walking in salamanders or during any other locomotor activity in previously studied tetrapods. The central neuronal mechanisms mediating locomotion in this primitive tetrapod are thus capable of considerable plasticity. PMID- 9405525 TI - Prolonged firing in motor units: evidence of plateau potentials in human motoneurons? AB - Serotonin (5-HT) and norepinephrine-dependent plateau potentials are found in spinal motoneurons in reduced turtle and cat preparations. Triggering the plateau potential by short-lasting synaptic excitation causes a prolonged self-sustained firing, which can be terminated by short-lasting synaptic inhibition. The presence of plateau potentials can also allow neurons to fire in a bistable manner, i.e., shifting between stable low and high firing frequencies. Such a bistable firing behavior has been found in soleus motor units in unrestrained rats. In the present study single motor-unit activity was recorded from low threshold units in human soleus and tibialis anterior muscles to evaluate whether a bistable firing behavior and/or prolonged firing could be evoked. Vibration of the homonymous muscle tendon (30-100 Hz, 2-10 s) was used as excitatory input to the motoneuron pool. Brief excitation while the muscle was electrically silent induced firing during the vibration and sometimes recruited units into prolonged stable firing outlasting the vibratory stimulus. However, a bistable firing behavior, i.e., vibration-induced maintained shifts between two stable levels of firing, could not be convincingly demonstrated. The reason for this was twofold. First, low-threshold human motor units tended to jump to a "preferred firing range" shortly after voluntary recruitment. This firing range was the same as when units were recruited from silence into prolonged firing by vibration. Below the preferred firing range, maintained firing was unstable and usually only possible when subjects were listening to the spike potentials or had visual force feedback. Second, vibration when units were firing in the preferred firing range caused a transient increase in firing frequency but no maintained frequency shifts. Recordings from pairs of motor units showed that short-lasting vibration could recruit one unit into prolonged firing, while a second unit, which already fired in its preferred firing range, only transiently increased its firing rate during the vibration. This suggests that the prolonged firing was not the result of an increase in the common synaptic drive to the motoneuron pool. We conclude that a bistable firing behavior as seen in intact rats is probably absent in human low-threshold motor units, but that prolonged firing could be seen in response to short-lasting excitation. This latter phenomenon is compatible with the existence of plateau potentials, which have to have a threshold close to the threshold for sodium spike generation. PMID- 9405526 TI - Heterologous expression of a P2x-purinoceptor in rat chromaffin cells detects vesicular ATP release. AB - A cloned P2x-purinoceptor was transiently expressed in single isolated rat adrenal chromaffin cells and evaluated for the detection of released ATP. After cytoplasmic injection of the P2x complementary RNA (cRNA; 4-24 h), application of ATP produced an inwardly rectifying current over the voltage range -130 to -10 mV as measured by the whole cell patch-clamp technique. The dose-response curve for ATP was sigmoidal with a 50% effective concentration of 18. 2 microM. Suramin, a P2x-antagonist, attenuated the ATP-induced current. Depolarizing voltage pulses to 0 mV or application of histamine, stimuli that trigger exocytosis, resulted in the appearance of suramin-sensitive spontaneous transient inward currents (at -60 mV) that resembled excitatory postsynaptic currents although they were slower in time course. Concurrent detection of catecholamine release with a carbon fiber electrode often showed coincidence of the amperometric current with the synaptic currentlike events suggesting that ATP and catecholamines were released from the same vessicle. These data demonstrate that expression of a P2x-purinoceptor in chromaffin cells produces a functional autoreceptor capable of detecting vesicular release of ATP. In combination with carbon fiber amperometry, simultaneous vesicular release of two neurotransmitters from a single chromaffin cell could be monitored. The P2x-purinoceptor, however, produced a regenerative effect on release apparently resulting from the high Ca2+ permeability of the receptor. Thus modification of the P2x-purinoceptor would be required before the system could be applied to examining processes involved in stimulus-release coupling. PMID- 9405527 TI - Stability of motor-unit force thresholds in the decerebrate cat. AB - To further test the hypothesis that some fixed property of motoneurons determines their recruitment order, we quantified the variation in force threshold (FT) for motoneurons recruited in muscle stretch reflexes in the decerebrate cat. Motor axons supplying the medial gastrocnemius (MG) muscle were penetrated with micropipettes and physiological properties of the motoneuron and its muscle fibers, i.e., the motor unit, were measured. FT, defined as the amount of MG force produced when the isolated motor unit was recruited, was measured from 20 to 93 consecutive stretch trials for 29 motor units. Trials were selected for limited variation in base force and rate of rise of force, which have been shown to covary with FT, and in peak stretch force, which gives some index of motor pool excitability. Under these restricted conditions, large variation in FT would have been inconsistent with the hypothesis. Analysis of the variation in FT employed the coefficient of variation (CV), because of the tendency for FT variance and mean to increase together. We found that CV was distributed with a median value of 10% and with only 2 of 29 units exceeding 36%. Some of this variation was associated with measurement error and with intertrial fluctuations in base, peak, and the rate of change of muscle force. CV was not significantly correlated with motor-unit axonal conduction velocity, contraction time, or force. In three cases FT was measured simultaneously from two motor units in the same stretch trials. Changes in recruitment order were rarely observed (5 of 121 stretch trials), even when FT ranges for units in a pair overlapped. We suggest that the large variation in recruitment threshold observed in some earlier studies resulted not from wide variation in the recruitment ranking of motoneurons within one muscle, but rather from variation in the relative activity of different pools of motoneurons. Our findings are consistent with the hypothesis that recruitment order is determined by some fixed property of alpha motoneurons and/or by some unvarying combination of presynaptic inputs that fluctuate in parallel. PMID- 9405528 TI - Topography and reciprocal activity of cerebellar Purkinje cells in the uvula nodulus modulated by vestibular stimulation. AB - In the rabbit uvula-nodulus, vestibular and optokinetic information is mapped onto parasagittal zones by climbing fibers. These zones are related functionally to different pairs of vertical semicircular canals, otolithic inputs and horizontal optokinetic inputs. Vestibular stimulation restricted to one of these zones modulates climbing fiber responses (CFRs). Within each of these zones, simple spikes (SSs) are modulated reciprocally with CFRs. In rabbits anesthetized with chloralose-urethan, we have used vestibular and optokinetic stimulation to evoke CFRs within a parasagittal zone while recording from Purkinje cells in adjacent zones. We have examined whether the CFRs evoked by vestibular stimulation in one zone influence the SSs of an adjacent zone. CFRs and SSs were recorded during roll vestibular stimulation. The orientation of the head of the rabbit with respect to the axis of rotation was varied systematically so that a climbing fiber null plane could be determined. This null plane was the orientation of the head about the vertical axis at which no modulation of the CFR was observed during rotation about the longitudinal axis of the vestibular rate table. In the left uvula-nodulus, a medial sagittal strip extending through all the folia contained Purkinje cells with CFRs that had optimal planes of stimulation coplanar with the left posterior-right anterior semicircular canals (LPC-RAC). Lateral to this strip was a strip of Purkinje cells with CFRs that were characterized by optimal planes corresponding to stimulation of the left anterior-right posterior semicircular canals (LAC-RPC). SSs in Purkinje cells were modulated out of phase with CFRs from the same Purkinje cell. The depth of modulation of both CFRs and SSs was reduced during rotation in the climbing fiber "null plane". The depth of modulation of SSs was greatest when recorded from Purkinje cells located at the center of semicircular canal-related strip. We observed that 1) all folia of the uvula-nodulus receive vestibular climbing fiber inputs; 2) these climbing fiber inputs convey information from the vertical semicircular canals and otoliths but not the horizontal semicircular canals; 3) CFRs evoked in a particular sagittal zone do not influence SSs in adjacent zones; 4) modulation of a CFRs in a particular Purkinje cell can occur without modulation of SSs in the same Purkinje cell, although modulation of SSs was not observed in the absence of CFR modulation; and 5) modulation of SSs sometimes preceded that of CFRs in the same cell, implying that interneuronal pathways may contribute to SS modulation. Climbing fiber-driven Golgi cells, the inhibitory axon terminals of which end on granule cell dendrites in the classic glomerular synapse, may provide this interneuronal mechanism. PMID- 9405529 TI - Conditioning-related protection from acoustic injury: effects of chronic deefferentation and sham surgery. AB - The inner ear can be made less vulnerable to acoustic injury by a "conditioning" treatment involving exposure to a moderate-level acoustic stimulus before the acoustic overexposure. The present study was designed to explore the role of the olivocochlear (OC) system in this "protection." Guinea pigs were divided into a number of groups: some (trauma-only) were exposed to a traumatic noise for 4 h at 109 dB SPL; others (condition/trauma) were conditioned by daily exposure to the same noise at 85 dB SPL before the traumatic exposure. In OC-intact animals, the condition/trauma group showed significantly less permanent threshold shift (PTS) than the trauma-only group as measured via compound action potentials and distortion-product otoacoustic emissions (DPOAEs). Other animals with identical noise-exposure regimens underwent deefferentation surgery before the start of conditioning: the OC bundle (OCB) was cut in the brain stem, either at the midline (cutting the crossed OCB to both ears) or at the sulcus limitans (cutting all OC fibers to 1 side). Lesion success was quantified by measuring OC fascicles to the outer hair cell region in each ear. The results from the surgical groups showed that total loss of the OCB significantly increased the noise-induced PTS, whereas loss of the COCB only did not; that the conditioning exposure in deefferented animals increased, rather than decreased, the PTS from the traumatic exposure; and that animals undergoing sham surgery (brain stem cuts that failed to transect the OCB) appeared protected whether or not they received the conditioning noise exposure. The latter result suggests that conditioning-related protection may arise from a generalized stress response, which can be elicited by noise exposure, brain surgery, or a variety of other means. The former results make an OC role in the conditioning process, per se, difficult to assess, given the large effects of OC activity on general acoustic vulnerability. PMID- 9405530 TI - Mechanisms of electrical coupling between pyramidal cells. AB - Direct electrical coupling between neurons can be the result of both electrotonic current transfer through gap junctions and extracellular fields. Intracellular recordings from CA1 pyramidal neurons of rat hippocampal slices showed two different types of small-amplitude coupling potentials: short-duration (5 ms) biphasic spikelets, which resembled differentiated action potentials and long duration (>20 ms) monophasic potentials. A three-dimensional morphological model of a pyramidal cell was employed to determine the extracellular field produced by a neuron and its effect on a nearby neuron resulting from both gap junctional and electric field coupling. Computations were performed with a novel formulation of the boundary element method that employs triangular elements to discretize the soma and cylindrical elements to discretize the dendrites. An analytic formula was derived to aid in computations involving cylindrical elements. Simulation results were compared with biological recordings of intracellular potentials and spikelets. Field effects produced waveforms resembling spikelets although of smaller magnitude than those recorded in vitro. Gap junctional electrotonic connections produced waveforms resembling small-amplitude excitatory postsynaptic potentials. Intracellular electrode measurements were found inadequate for ascertaining membrane events because of externally applied electric fields. The transmembrane voltage induced by the electric field was highly spatially dependent in polarity and wave shape, as well as being an order of magnitude larger than activity measured at the electrode. Membrane voltages because of electrotonic current injection across gap junctions were essentially constant over the cell and were accurately depicted by the electrode. The effects of several parameters were investigated: 1) decreasing the ratio of intra to extracellular conductivity reduced the field effects; 2) the tree structure had a major impact on the intracellular potential; 3) placing the gap junction in the dendrites introduced a time delay in the gap junctional mediated electrotonic potential, as well as deceasing the potential recorded by the somatic electrode; and 4) field effects decayed to one-half of their maximum strength at a cell separation of approximately 20 micron. Results indicate that the in vitro measured spikelets are unlikely to be mediated by gap junctions and that a spikelet produced by the electric field of a single source cell has the same waveshape as the measured spikelet but with a much smaller amplitude. It is hypothesized that spikelets are a manifestation of the simultaneous electric field effects from several local cells whose action potential firing is synchronized. PMID- 9405531 TI - Recurrent inhibitory interneurons of the Rabbit's lateral posterior-pulvinar complex. AB - We recorded from 118 neurons in the visual sector of the thalamic reticular nucleus (TRN) in anesthetized rabbits. Cells were identified by their location and characteristic burst responses to stimulation of the primary visual cortex (Cx) and optic chiasm (OX) and were classified into two groups. Type I cells had relatively short latencies from both OX and Cx stimulation, and the latency from OX was always longer than from Cx. In contrast, type II cells had much longer latencies after OX and Cx stimulation, and the latency from OX was always shorter than from Cx. Type I cells were located in the dorsal part of TRN, whereas type II cells were located in the ventral part of TRN. The physiological properties and location of type I TRN cells indicate that they are recurrent inhibitory interneurons of the dorsal lateral geniculate nucleus (LGN). Type II TRN cells most likely function as recurrent inhibitory interneurons for the lateral posterior nucleus-pulvinar complex (LP) because they could be activated antidromically by LP stimulation and orthodromically activated via axonal collaterals of LP cells. Type II TRN cells exhibited a prolonged depression after Cx or OX stimulation. Intracellular recordings showed that a prolonged inhibitory postsynaptic potential was evoked by Cx or OX stimulation. Therefore, these recurrent interneurons of LP, type II cells form mutual inhibitory connections just like those recurrent interneurons of LGN, type I cells. Our data suggest that the geniculocortical and extrageniculate visual pathways have similar recurrent inhibitory circuits. PMID- 9405532 TI - Voltage-dependent conductances in cephalopod primary sensory hair cells. AB - Cephalopods, such as sepia, squid, and octopus, show a well-developed and sophisticated control of balance particularly during prey capture and escape behaviors. There are two separate areas of sensory epithelium in cephalopod statocysts, a macula/statolith system, which detects linear accelerations (gravity), and a crista/cupula system, which detects rotational movements. The aim of this study is to characterize the ionic conductances in the basolateral membrane of primary sensory hair cells. These were studied using a whole cell patch-clamp technique, which allowed us to identify five ionic conductances in the isolated primary hair cells; an inward sodium current, an inward calcium current, and three potassium outward currents. These outward currents were distinguishable on the basis of their voltage-dependence and pharmacological sensitivities. First, a transient outward current (IA) was elicited by depolarizing voltage steps from a holding potential of -60 mV, was inactivated by holding the cell at -40 mV, and was blocked by 4-aminopyridine. A second, voltage sensitive, outward current with a sustained time course was identified. This current was not blocked by 4-aminopyridine nor inactivated at a holding potential of -40 mV and hence could be separated from IA using these protocols. A third outward current that depended on Ca2+ entry for its activation was detected, this current was identified by its sensitivity to Ca2+ channel blockers such as Co2+ and Cd2+ and by the N-shaped profile of its current-voltage curve. Inward currents were studied using cesium aspartate solution in the pipette to block the outward currents. Two inward currents were observed in the primary sensory hair cells. A fast transient inward current, which is presumably responsible for spike generation. This inward current appeared as a rapidly activating inward current; this was strongly voltage dependent. Three lines of evidence suggest that this fast transient inward current is a Na+ current (INa). First, it was blocked by tetrodotoxin (TTX); second, it also was blocked by Na+-free saline; and third, it was inactivated when primary hair cells were held at a potential more than -40 mV. The sustained inward current was not affected by TTX and was increased in amplitude 5 min after equimolar Ba2+ replaced Ca2+ as a charge carrier. This inward current also was blocked after external application of 2 mmol/l Co2+ or Cd2+. Furthermore, this current was reduced significantly in a dose-dependent manner by nifedipine, suggesting that it is an L-type Ca2+ current (ICa). PMID- 9405533 TI - Functional analysis of the sensory motor pathway of resistance reflex in crayfish. I. Multisensory coding and motor neuron monosynaptic responses. AB - An in vitro preparation of the fifth thoracic ganglion of the crayfish was used to study in detail the negative feedback loop involved in the control of passive movements of the leg. Release-sensitive primary afferents of from the coxo basipodite chordotonal organ (CBCO), a proprioceptor whose strand is released by upward movement of the leg, monosynaptically connect to depressor motor neurons (Dep MNs). Extracellular identification of sensory units from the CBCO neurogram allowed us to determine the global coding of a sine-wave movement, imposed from the most released position of the CBCO strand. Intracellular recordings from sensory terminals (CBTs) and ramp movement stimulations applied to the CBCO strand allowed us to characterize two groups of release-sensitive CBCO fibers. The first group, divided into two subgroups (phasic and phaso-tonic), is characterized by discontinuous firing patterns: phasic CBTs fired exclusively during release movements; phaso-tonic CBTs displayed both a phasic firing and a tonic discharge during the more released plateaus. The second group was continuously firing whatever the movement, with higher frequencies during the release phase of the movement stimulation. All CBTs displayed a marked sensitivity for release movements while only the phaso-tonic ones showed a clear sensitivity to maintained positions. Surprisingly, no pure tonic sensory fibers were encountered. Systematic intracellular recordings from all resistant Dep MNs, performed in high divalent cation saline, allowed us to describe two shapes of monosynaptic resistance reflex responses. A phasic response was characterized by bursts of excitatory postsynaptic potentials (EPSPs) occurring exclusively during CBCO strand release movements. A phaso-tonic response was characterized by a progressive depolarization occurring all along the release phase of the stimulation: during maintained released positions, the amplitude of the sustained depolarization was position dependent; in addition, each release movement produced a phasic burst of EPSPs in the MN. The parallel study of the Dep MN properties failed to point out any correlation between the type of reflex response recorded from the MN and the MN intrinsic properties, which would indicate that the type of MN response is entirely determined by the afferent messages it receives. PMID- 9405534 TI - Functional analysis of the sensory motor pathway of resistance reflex in crayfish. II. Integration Of sensory inputs in motor neurons. AB - The in vitro preparation of the fifth thoracic ganglion of the crayfish was used to analyze the connections supporting the monosynaptic reflex responses recorded from the depressor motor neurons (Dep MNs). Dep MNs are directly connected by the release-sensitive afferents from a proprioceptor, the coxo-basipodite chordotonal organ (CBCO), which is released by upward movements of the leg. Sine-wave movements, applied to the CBCO strand from the most released position, allowed us to stimulate the greatest part of release-sensitive CBCO fibers. Systematic intracellular recordings from all Dep MNs performed in high divalent cation saline allowed us to determine the connections between CBCO afferents and their postsynaptic Dep MNs: it highlighted the sequential activation of the different Dep MNs involved in the monosynaptic reflex. The convergence of different sensory afferents onto a given Dep MN, and the divergence of a given sensory afferent onto several Dep MNs illustrates the complexity of the sensory-motor reflex loops involved in the control of locomotion and posture. Electrophysiological experiments and simulations were performed to analyze the mechanisms by which Dep MNs integrate the large amount of sensory input that they receive. Paired intracellular recording experiments demonstrated that postsynaptic response shapes characteristic of both phasic and phaso-tonic afferents could be induced by varying the presynaptic firing frequency, whatever the postsynaptic Dep MN. Compartment model simulations were used to analyze the role of the sensory-motor synapse characteristics in the summation properties of postsynaptic MN. They demonstrated the importance of the postsynaptic compartment geometry, because large postsynaptic compartments allowed to generate greater excitatory postsynaptic potential (EPSP) summations than small ones. The results presented show that velocity information is the most effective to elicit large compound EPSPs in MNs. We therefore suggest that the negative feedback reflex is mainly based on the detection of leg movements. PMID- 9405535 TI - Activation and recovery of the PGE2-mediated sensitization of the capsaicin response in rat sensory neurons. AB - Pro-inflammatory prostaglandins are known to enhance the sensitivity of sensory neurons to various modalities of stimulation, including the excitatory chemical agent, capsaicin. In this report, we examined the capacity of prostaglandin E2 (PGE2) to enhance the capsaicin response recorded from sensory neurons isolated from embryonic rats and grown in culture. Previous work demonstrated that the cyclic adenosine 3',5'-monophosphate pathway mediates initiation of the PGE2 induced sensitization, however, little is known about the pathways regulating the recovery from sensitization. Therefore, we examined the neuronal transduction cascades that control the duration of sensitization. Treatment with PGE2 enhanced the capsaicin-evoked current by two- to threefold, however, this sensitization was transient even in the continued presence of prostaglandin. The duration of sensitization produced by PGE2 was related inversely to the extracellular Ca2+ concentration with the shortest recovery times observed in cells exposed to 2 mM Ca2+-Ringer. Inclusion of the Ca2+ chelator, bis-(o-aminophenoxy)-N, N,N',N' tetraacetic acid, in the recording pipette greatly lengthened the period of sensitization. Pretreatment with either the nitric oxide synthase inhibitor, nitro-L-arginine methyl ester (L-NAME), or the inhibitor of the cyclic guanosine 3', 5'-monophosphate (GMP)-dependent protein kinase, KT-5823, before the application of PGE2 increased the duration of sensitization even in the presence of 2 mM Ca2+. In contrast, after attaining maximal sensitization in 2 mM Ca2+ Ringer containing L-NAME, the addition of either nitric oxide donors (3 morpholinosydnonimine or s-nitroso-n-acetylpenicillamine) or 8-Br-cyclic GMP led to a rapid decrease in the level of sensitization. In the absence of sensitization, nitric oxide-cyclic GMP modulating agents had no effect on the capsaicin-evoked current. Therefore, these results suggest that capsaicin-induced elevations in intracellular Ca2+ levels lead to an enhanced production of cyclic GMP, via the nitric oxide pathway, that ultimately activates cyclic GMP-dependent protein kinase. This protein kinase inactivates or terminates the sensitization produced by PGE2 by an as yet unidentified mechanism. PMID- 9405536 TI - Temporal summation of C-fiber afferent inputs: competition between facilitatory and inhibitory effects on C-fiber reflex in the rat. AB - Long-lasting facilitations of spinal nociceptive reflexes resulting from temporal summation of nociceptive inputs have been described on many occasions in spinal, nonanesthetized rats. Because noxious inputs also trigger powerful descending inhibitory controls, we investigated this phenomenon in intact, halothane anesthetized rats and compared our results with those obtained in other preparations. The effects of temporal summation of nociceptive inputs were found to be very much dependent on the type of preparation. Electromyographic responses elicited by single square-wave electrical shocks (2 ms, 0.16 Hz) applied within the territory of the sural nerve were recorded in the rat from the ipsilateral biceps femoris. The excitability of the C-fiber reflex recorded at 1.5 times the threshold (T) was tested after 20 s of electrical conditioning stimuli (2 ms, 1 Hz) within the sural nerve territory. During the conditioning procedure, the C fiber reflex was facilitated (wind-up) in a stimulus-dependent fashion in intact, anesthetized animals during the application of the first seven conditioning stimuli; thereafter, the magnitude of the responses reached a plateau and then decreased. Such a wind-up phenomenon was seen only when the frequency of stimulation was 0.5 Hz or higher. In spinal, unanesthetized rats, the wind-up phenomenon occurred as a monotonic accelerating function that was obvious during the whole conditioning period. An intermediate picture was observed in the nonanesthetized rat whose brain was transected at the level of the obex, but the effects of conditioning were profoundly attenuated when such a preparation was anesthetized. In intact, anesthetized animals the reflex was inhibited in a stimulus-dependent manner during the postconditioning period. These effects were not dependent on the frequency of the conditioning stimulus. Such inhibitions were blocked completely by transection at the level of the obex, and in nonanesthetized rats were then replaced by a facilitation. A similar long-lasting facilitation was seen in nonanesthetized, spinal rats. It is concluded that, in intact rats, an inhibitory mechanism counteracts the long-lasting increase of excitability of the flexor reflex seen in spinal animals after high-intensity, repetitive stimulation of C-fibers. It is suggested that supraspinally mediated inhibitions also participate in long term changes in spinal cord excitability after noxious stimulation. PMID- 9405537 TI - Glutamate microstimulation of local inhibitory circuits in the supraoptic nucleus from rat hypothalamus slices. AB - The hypothesis of a local inhibitory input to the hypothalamic supraoptic nucleus was tested with combined glutamate microstimulation and whole cell patch-clamp recordings in slices from rat hypothalamus. Synaptic activity in supraoptic magnocellular neuroendocrine cells (MNCs) was monitored and glutamate microdrops were applied in the perinuclear region of the supraoptic nucleus to evoke firing of action potentials in putative presynaptic inhibitory cells. The effect of glutamate microdrops applied in the perinuclear region was tested on 57 supraoptic MNCs. In control conditions, spontaneous excitatory (EPSCs) and inhibitory (IPSCs) postsynaptic currents were observed at resting membrane potential in all MNCs tested. Glutamate microstimulation evoked an abrupt increase in the frequency and size of spontaneous IPSCs in eight MNCs. Forty-nine MNCs did not show any change in the inhibitory synaptic input. Microapplication of glutamate in the periphery of the supraoptic nucleus did not modify the amplitude or the frequency of spontaneous EPSCs in any of the 57 MNCs tested. In the group of eight MNCs that responded to glutamate microstimulation by an increase in inhibitory input, two types of responses were observed. Four MNCs showed an increase in both size and frequency of spontaneous IPSCs through the entire range of amplitude. In the other four MNCs, local glutamate stimulation produced a dramatic increase in the size of IPSCs and a lesser increase in the frequency of the smaller IPSCs. The potential effect of the glutamate-evoked increase in inhibitory input on the firing activity of MNCs was tested in current clamp conditions. Intracellular current injection was applied to evoke firing of action potentials in six MNCs that had responded to local glutamate microstimulation by an increase in inhibitory input. Glutamate microdrop applications inhibited the evoked action potential firing in all six cells. These results suggest 1) that local inhibitory interneurons are present in the periphery of the supraoptic nucleus, 2) that they contain functional glutamate receptors, 3) that they form inhibitory synapses with supraoptic MNCs, and 4) that activation of these interneurons inhibits firing in MNCs. These results support the hypothesis that local inhibitory interneurons play a important role in the firing activity of supraoptic MNCs. PMID- 9405538 TI - Insensitivity of V1 complex cell responses to small shifts in the retinal image of complex patterns. AB - An important role for neurons in the early visual system is to convey information about the structure of visual stimuli. However, neuronal responses show substantial variation across presentations of the same stimulus. In awake monkeys, it has been assumed that a great deal of this variation is related to the scatter in eye position (inducing scatter in the retinal position of the stimulus). Here we investigate the implied consequence of this assumption, i.e., that the scatter variation in eye position degrades the decodability of the neural response. We recorded from 50 complex cells in primary visual cortex of fixating monkeys while different complex stimuli were presented. Three types of retinal shifts were considered: natural scatter in the fixation, systematic fixation point shift, and systematic stimulus position shift. The stimulus pattern accounts for >50% of the response variance, always six times that accounted for by the scatter in eye position during fixation. The retinal location of a stimulus had to be shifted by 10-12 min of arc, an amount almost two times larger than the smallest picture element, before the responses changed systematically. Nonetheless, changes of the stimulus at the single pixel level often gave rise to discriminable responses. Thus complex cells convey information about the spatial structure of a stimulus, independent of rigid stimulus displacements on the order of the receptive field size or smaller. PMID- 9405539 TI - Experimental and modeling study of Na+ current heterogeneity in rat nodose neurons and its impact on neuronal discharge. AB - This paper is a combined experimental and modeling study of two fundamental questions surrounding the functional characteristics of Na+ currents in nodose sensory neurons. First, when distinctly different classes of Na+ currents are expressed in the same neuron, is there a significant difference in the intrinsic biological variability associated with the voltage- and time-dependent properties of these currents? Second, in what manner can such variability in functional properties impact the discharge characteristics of these neurons? Here, we recorded the whole cell Na+ currents in acutely dissociated rat nodose sensory neurons using the patch-clamp technique. Two general populations of neurons were observed. A-type neurons (n = 20) expressed a single rapidly inactivating tetrodotoxin-sensitive (TTX-S) Na+ current. C-type neurons (n = 87) coexpressed this TTX-S current along with a slowly inactivating TTX-resistant (TTX-R) Na+ current. The TTX-S currents in both cell types had submillisecond rates of activation at room temperature with thresholds near -50 mV. The TTX-R current exhibited about the same rates of activation but required potentials 20-30 mV more depolarized to reach threshold. Over the same clamp voltages the rates of inactivation for the TTX-R current were three to nine times slower than those for the TTX-S current. However, the TTX-R current recovered from complete inactivation at a rate 10-20 times faster than the TTX-S current (10 ms as compared with 100-200 ms). Across the population of neurons studied the TTX-S data formed a relatively tight statistical distribution, exhibiting low standard deviations across all measured voltage- and time-dependent properties. In contrast, the same pooled measurements on the TTX-R data exhibited standard deviations that were 3-10 times larger. The statistical profiles of the voltage- and time-dependent properties of these currents then were used as a physiological guide to adjust the relevant parameters of a mathematical model of nodose sensory neurons previously developed by our group (). Here, we show how the relative expression of TTX-S and TTX-R Na+ currents and the differences in their apparent biological variability can shape the regenerative discharge characteristics and action potential waveshapes of sensory neurons. We propose that the spectrum of variability robust reactivation characteristics of the TTX-R current are important determinants in establishing the heterogeneous stimulus-response characteristics often observed across the general population of C-type sensory neurons. PMID- 9405540 TI - Amine modulation of glutamate responses from pyloric motor neurons in lobster stomatogastric ganglion. AB - The amines dopamine (DA), serotonin (5-HT), and octopamine (Oct) each elicit a distinctive motor pattern from a quiescent pyloric network in the lobster stomatogastric ganglion (STG). We previously have demonstrated that these amines alter the synaptic strength at multiple, distributed sites within the pyloric network that could contribute to the amine-induced motor patterns. Here, we examined the postsynaptic contribution to these changes in synaptic strength by determining how the amines modify responses of pyloric motor neurons to glutamate (Glu), one of the network transmitters, applied iontophoretically into the STG neuropil. Dopamine reduced the Glu responses of the pyloric dilator (PD), ventricular dilator (VD), and inferior cardiac (IC) neurons and enhanced the Glu responses of the lateral pyloric (LP) and pyloric constrictor (PY) neurons. The only effect of 5-HT was to reduce the Glu response of the VD neuron. Oct enhanced the Glu responses of the LP and PY neurons but did not affect the PD, VD, and IC responses. We also examined amine effects on the depolarizing responses to iontophoresed acetylcholine (ACh) in the PD and VD and found that they paralleled the amine effects on Glu responses in these neurons. This suggests that amine modulation of PD and VD responses to Glu and ACh may be explained by general changes in the ionic conductance of these neurons. We compare our results with our earlier work describing amine effects on synaptic strength and input resistance to show that amines act at both pre- and postsynaptic sites to modify graded synaptic transmission in the pyloric network. PMID- 9405541 TI - Mathematical model of the human jaw system simulating static biting and movements after unloading. AB - When the resistance to a forceful isometric bite is suddenly removed in unloading experiments, the bite force drops to zero and the mandible reaches a constant velocity. This occurs at an initial bite force of 100 N after approximately 12 ms when the incisors have moved 4.5 mm. Reflex activity is far too slow to limit the velocity at impact. To explore the influence of other factors (cocontraction, force-length properties, and force-velocity properties of the muscles) on the velocity at impact, a numerical forward dynamic model of the jaw system is formulated. Unloading experiments in different experimental conditions were simulated with the model. Most parameter values of the model are based on physiological data, both from literature and a data basis from a human cadaver study. Other parameter values were found by optimally fitting the model results to data from the unloading experiments. The model analysis shows that the limitation of the jaw velocity mainly may be due to the force-velocity properties of the jaw-closing muscles. Force-length properties of the jaw muscles hardly contribute to the impact velocity. The compliance of tendinous sheets in the jaw muscles is unfavorable for the reduction in impact velocity, whereas cocontraction of jaw-opening and -closing muscles helps to limit impact velocity. The force-velocity properties of the muscles provide a quick mechanism for dealing with unexpected closing movements and so avoid damage to the dental elements. PMID- 9405542 TI - Three-dimensional analysis of vestibular efferent neurons innervating semicircular canals of the gerbil. AB - Anterograde labeling techniques were used to examine peripheral innervation patterns of vestibular efferent neurons in the crista ampullares of the gerbil. Vestibular efferent neurons were labeled by extracellular injections of biocytin or biotinylated dextran amine into the contralateral or ipsilateral dorsal subgroup of efferent cell bodies (group e) located dorsolateral to the facial nerve genu. Anterogradely labeled efferent terminal field varicosities consist mainly of boutons en passant with fewer of the terminal type. The bouton swellings are located predominately in apposition to the basolateral borders of the afferent calyces and type II hair cells, but several boutons were identified close to the hair cell apical border on both types. Three-dimensional reconstruction and morphological analysis of the terminal fields from these cells located in the sensory neuroepithelium of the anterior, horizontal, and posterior cristae were performed. We show that efferent neurons densely innervate each end organ in widespread terminal fields. Subepithelial bifurcations of parent axons were minimal, with extensive collateralization occurring after the axons penetrated the basement membrane of the neuroepithelium. Axonal branching ranged between the 6th and 27th orders and terminal field collecting area far exceeds that of the peripheral terminals of primary afferent neurons. The terminal fields of the efferent neurons display three morphologically heterogeneous types: central, peripheral, and planum. All cell types possess terminal fields displaying a high degree of anisotropy with orientations typically parallel to or within +/-45 degrees of the longitudinal axis if the crista. Terminal fields of the central and planum zones predominately project medially toward the transverse axis from the more laterally located penetration of the basement membrane by the parent axon. Peripheral zone terminal fields extend predominately toward the planum semilunatum. The innervation areas of efferent terminal fields display a trend from smallest to largest for the central, peripheral, and planum types, respectively. Neurons that innervate the central zone of the crista do not extend into the peripheral or planum regions. Conversely, those neurons with terminal fields in the peripheral or planum regions do not innervate the central zone of the sensory neuroepithelium. The central zone of the crista is innervated preferentially by efferent neurons with cell bodies located in the ipsilateral group e. The peripheral and planum zones of the crista are innervated preferentially by efferent neurons with cell bodies located in the contralateral group e. A model incorporating our anatomic observations is presented describing an ipsilateral closed-loop feedback between ipsilateral efferent neurons and the periphery and an open-loop feed-forward innervation from contralateral efferent neurons. A possible role for the vestibular efferent neurons in the modulation of semicircular canal afferent response dynamics is proposed. PMID- 9405543 TI - Pharmacological characterization of Na+ influx via voltage-gated Na+ channels in spinal cord astrocytes. AB - Spinal cord astrocytes display a high density of voltage-gated Na+ channels. To study the contribution of Na+ influx via these channels to Na+ homeostasis in cultured spinal cord astrocytes, we measured intracellular Na+ concentration ([Na+]i) with the fluorescent dye sodium-binding benzofuran isophthalate. Stellate and nonstellate astrocytes, which display Na+ currents with different properties, were differentiated. Baseline [Na+]i was 8.5 mM in these cells and was not altered by 100 microM tetrodotoxin (TTX). Inhibition of Na+ channel inactivation by veratridine (100 microM) evoked a [Na+]i increase of 47.1 mM in 44% of stellate and 9.7 mM in 64% of nonstellate astrocytes. About 30% of cells reacted to veratridine with a [Na+]i decrease of approximately 2 mM. Qualitatively similar [Na+]i changes were caused by aconitine. The effects of veratridine were blocked by TTX, amplified by (alpha-)scorpion toxin and usually were readily reversible. Veratridine-induced [Na+]i increases were reduced upon membrane depolarization with elevated extracellular [K+]. Recovery to baseline [Na+]i was unaltered during blocking of K+ channels with 4-aminopyridine. [Na+]i increases evoked by the ionotropic non-N-methyl--aspartate receptor agonist kainate were not altered by TTX. Our results indicate that influx of Na+ via voltage- gated Na+ channels is not a prerequisite for glial Na+,K+-ATPase activity in spinal cord astrocytes at rest nor does it seem to be involved in [Na+]i increases evoked by kainate. During pharmacological inhibition of Na+ channel inactivation, however, Na+ channels can serve as prominent pathways of Na+ influx and mediate large perturbations in [Na+]i, suggesting that Na+ channel inactivation plays an important functional role in these cells. PMID- 9405544 TI - Analysis of primate IBN spike trains using system identification techniques. I. Relationship To eye movement dynamics during head-fixed saccades. AB - The dynamic behavior of primate (Macaca fascicularis) inhibitory burst neurons (IBNs) during head-fixed saccades was analyzed by using system identification techniques. Neurons were categorized as IBNs on the basis of their anatomic location as well as by their activity during horizontal head-fixed saccadic and smooth pursuit eye movements and vestibular nystagmus. Each IBN's latency or "dynamic lead time" (td) was determined by shifting the unit discharge in time until an optimal fit to the firing rate frequency B(t) profile was obtained by using the simple model based on eye movement dynamics,B(t) = r + b1(t); where is eye velocity. For the population of IBNs, the dynamic estimate of lead time provided a significantly lower value than a method that used the onset of the first spike. We then compared the relative abilities of different eye movement based models to predict B(t) by using objective optimization algorithms. The most important terms for predicting B(t) were eye velocity gain (b1) and bias terms (r) mentioned above. The contributions of higher-order velocity, acceleration, and/or eye position terms to model fits were found to be negligible. The addition of a pole term [the time derivative of B(t)] in conjunction with an acceleration term significantly improved model fits to IBN spike trains, particularly when the firing rates at the beginning of each saccade [initial conditions (ICs)] were estimated as parameters. Such a model fit the data well (a fit comparable to a linear regression analysis with a R2 value of 0.5, or equivalently, a correlation coefficient of 0.74). A simplified version of this model [B(t) = rk + b1(t)], which did not contain a pole term, but in which the bias term (rk) was estimated separately for each saccade, provided nearly equivalent fits of the data. However, models in which ICs or rks were estimated separately for each saccade contained too many parameters to be considered as useful models of IBN discharges. We discovered that estimated ICs and rks were correlated with saccade amplitude for the majority of short-lead IBNs (SLIBNs; 56%) and many long-lead IBNs (LLIBNs; 42%). This observation led us to construct a more simple model that included a term that was inversely related to the amplitude of the saccade, in addition to eye velocity and constant bias terms. Such a model better described neuron discharges than more complex models based on a third-order nonlinear function of eye velocity. Given the small number of parameters required by this model (only 3) and its ability to fit the data, we suggest that it provides the most valuable description of IBN discharges. This model emphasizes that the IBN discharges are dependent on saccade amplitude and implies further that a mechanism must exist, at the motoneuron (MN) level, to offset the effect of the bias and amplitude-dependent terms. In addition, we did not find a significant difference in the variance accounted for by any of the downstream models tested for SLIBNs versus LLIBNs. Therefore we conclude that the eye movement signals encoded dynamically by SLIBNs and LLIBNs are similar in nature. Put another way, SLIBNs are not closer, dynamically, to MNs than LLIBNs. PMID- 9405545 TI - Analysis of primate IBN spike trains using system identification techniques. II. Relationship to gaze, eye, and head movement dynamics during head-free gaze shifts. AB - We have investigated the relationships among the firing frequency B(t) of inhibitory burst neurons (IBNs) and the metrics and dynamics of the eye, head, and gaze (eye + head) movements generated during voluntary combined eye-head gaze shifts in monkey. The same IBNs were characterized during head-fixed saccades in our first of three companion papers. In head-free gaze shifts, the number of spikes (NOS) in a burst was, for 82% of the neurons, better correlated with gaze amplitude than with the amplitude of either the eye or head components of the gaze shift. A multiple regression analysis confirmed that NOS was well correlated to the sum of head and eye amplitudes during head-free gaze shifts. Furthermore, the mean slope of the relationship between NOS and gaze amplitude was significantly less for head-free gaze shifts than for head-fixed saccades. NOS is a global parameter. To refine we used system identification techniques to evaluate a series of dynamic models in which IBN spike trains were related to gaze or eye movements. We found that gaze- and eye-based models predicted the discharges of IBNs equally well. However, the bias values required by gaze-based models were comparable to those required in our head-fixed models whereas those required by eye-based models were significantly larger. The difference in biases between gaze- and eye-based models was very strongly correlated to the mean head velocity () during gaze shifts [R = -0.93 +/- 0.15 (SD)]. This result suggested that the increased bias required by the eye-based models reflected an unmodeled input onto these cells. To pursue this argument further we investigated a series of dynamic models that included both eye velocity () and terms and this confirmed the importance of these two terms. As in our head-fixed analysis of companion paper I, the most valuable model formulation also included an eye saccade amplitude term (DeltaE) and was given by B(t) = r0 + r1DeltaE + b1 + g1 where r0, r1, b1, and g1 are constants. The amplitude of the head velocity coefficient was significantly less than that of the eye velocity coefficient. Furthermore, in our population long-lead IBNs tended to have a smaller head velocity coefficients than short-lead IBNs. We conclude that during head-free gaze shifts, the head velocity signal carried to the abducens nucleus by primate excitatory burst neurons (EBNs; if EBNs and IBNs carry similar signals) must be offset by other premotor cells. PMID- 9405546 TI - Analysis of primate IBN spike trains using system identification techniques. III. Relationship To motor error during head-fixed saccades and head-free gaze shifts. AB - The classic model of saccade generation assumes that the burst generator is driven by a motor-error signal, the difference between the actual eye position and the final "desired" eye position in the orbit. Here we evaluate objectively, using system identification techniques, the dynamic relationship between motor error signals and primate inhibitory burst neuron (IBN) discharges (upstream analysis). The IBNs presented here are the same neurons whose downstream relationships were characterized during head-fixed saccades and head-free gaze shifts in our companion papers. In our analysis of head-fixed saccades we determined how well IBN discharges encode eye motor error (epsilone) compared with downstream saccadic eye movement dynamics and whether long-lead IBN (LLIBN) discharges encode epsilone better than short-lead IBNs (SLIBNs), given that it is commonly assumed that short-lead burst neurons (BNs) are closer than long-lead BNs to the motor output and thus further from the epsilone signal. In the epsilone-based models tested, IBN firing frequency B(t) was represented by one of the following: 1) model 1u, a nonlinear function of epsilone; 2) model 2u, a linear function of epsilone [B(t) = rk + a1epsilone(t)] where the bias term rk was estimated separately for each saccade; 3) model 3u, a version of model 2u wherein the bias term was a function of saccade amplitude; or 4) model 4u, a linear function of epsilone with an added pole term (the derivative of firing rate). Models based on epsilone consistently produced worse predictions of IBN activity than models of comparable complexity based on eye movement dynamics (e.g., eye velocity). Hence, the simple two parameter downstream model 2d [B(t) = r + b1(t)] was much better than any upstream (epsilone-based) model with a comparable number of parameters. The link between B(t) and epsilone is due primarily to the correlation between the declining phases of B(t) and epsilone; motor-error models did not predict well the rising phase of the discharge. We could improve substantially the performance of upstream models by adding an e term. Because e = -, this process was equivalent to incorporating terms into upstream models thereby erasing the distinction between upstream and downstream analyses. Adding an e term to the upstream models made them as good as downstream ones in predicting B(t). However, the epsilone term now became redundant because its removal did not affect model accuracy. Thus, when is available as a parameter, epsilone becomes irrelevant. In the head-free monkey the ability of upstream models to predict IBN firing during head-free gaze shifts when gaze, eye, or head motor-error signals were model inputs was poor and similar to the upstream analysis of the head-fixed condition. We conclude that during saccades (head-fixed) or gaze shifts (head-free) the activity of both SLIBNs and LLIBNs is more closely linked to downstream events (i.e., the dynamics of ongoing movements) than to the coincident upstream motor-error signal. Furthermore, SLIBNs and LLIBNs do not differ in their characteristics; the latter are not, as is usually hypothesized, closer to a motor-error signal than the former. PMID- 9405547 TI - Electrophysiological properties of rat pontine nuclei neurons In vitro. I. Membrane potentials and firing patterns. AB - We used a new slice preparation of rat brain stem to establish the basic membrane properties of neurons in the pontine nuclei (PN). Using standard intracellular recordings, we found that pontine cells displayed a resting membrane potential of -63 +/- 6 mV (mean +/- SD), an input resistance of 53 +/- 21 MOmega, a membrane time constant of 5.3 +/- 2.4 ms and were not spontaneously active. The current voltage relationship of most of the PN neurons showed the characteristics of inward rectification in both depolarizing and hyperpolarizing directions. A prominent feature of the firing of pontine neurons was a marked firing rate adaptation, which eventually caused the cells to cease firing. Several types of membrane conductances possibly contribute to this feature. For one, a medium and a slow type of afterhyperpolarization (AHP) control the pattern of firing. The medium AHP was partly susceptible to blockade of calcium influx, whereas it was abolished completely by blockade of potassium channels with tetraethylammonium, indicating that it is based on at least two conductances: a calcium-dependent and a calcium-independent one. The slow AHP was carried by potassium ions and could be blocked effectively by preventing calcium influx into the cell. It was present after single spikes but was strongest after a high-frequency spike train. Calcium entry into the cell was mediated by high-threshold calcium channels that were detected by the generation of calcium spikes under blockade of potassium channels. Furthermore, the early phase of the firing rate adaptation was shown to be related to the time course of a slow, tetrodotoxin (TTX)-sensitive, persistent sodium potential, which was activated already in the subthreshold range of membrane potentials. This potential was time dependent and imposed as a depolarizing "hump" with a maximum occurring in most cases between 50 and 100 ms after stimulus onset. In the suprathreshold range, it generated plateau potentials following fast spikes, if potassium channels were blocked. After the complete adaptation of the firing rate, PN neurons were observed to display irregular fluctuations of the membrane potential, which sometimes reached firing threshold thereby eliciting an irregular low-frequency spike train. As these fluctuations could be blocked with TTX, they probably are based on the persistent sodium currents. The opposing drive in hyperpolarizing direction may be provided by strong outward currents that generated a marked outward rectification in the current-voltage relationship under TTX. In conclusion, PN neurons show complex membrane properties that are reminiscent in many ways to cerebrocortical "regular firing" neurons. PMID- 9405548 TI - Electrophysiological properties of rat pontine nuclei neurons In vitro II. Postsynaptic potentials. AB - We investigated the postsynaptic responses of neurons of the rat pontine nuclei (PN) by performing intracellular recordings in parasagittal slices of the pontine brain stem. Postsynaptic potentials (PSPs) were evoked by brief (0.1 ms) negative current pulses (10-250 microA) applied to either the cerebral peduncle or the pontine tegmentum. First, excitatory postsynaptic potentials (EPSPs) could be evoked readily from peduncular stimulation sites. These EPSPs exhibited short latencies, a nonlinear increment in response to increased stimulation currents, and an unconventional dependency on the somatic membrane potential. Pharmacological blockade of the synaptic transmission using 6,7 dinitroquinoxaline-2, 3-dione and ,-2-amino-5-phosphonovaleric acid, selective antagonists of the alpha-amino-3-hydroxy-5-methyl-4-isoxazilepropionate- (AMPA) and the N-methyl--aspartate (NMDA)-type glutamate receptors, showed that these EPSPs were mediated exclusively by excitatory amino acids via both AMPA and NMDA receptors. Moreover, the pharmacological experiments indicated the existence of voltage-sensitive but NMDA receptor-independent amplification of EPSPs. Second, stimulations at peduncular and tegmental sites also elicited inhibitory postsynaptic potentials (IPSPs) in a substantial proportion of pontine neurons. The short latencies of all IPSPs argued against the participation of inhibitory interneurons. Their sensitivity to bicuculline and reversal potentials around -70 mV suggested that they were mediated by gamma-aminobutyric acid-A (GABAA) receptors. In addition to single PSPs, sequences consisting of two to four distinct EPSPs could be recorded after stimulation of the cerebral peduncle. Most remarkably, the onset latencies of the following EPSPs were multiples of the first one indicating the involvement of intercalated synapses. Finally, we used the classic paired-pulse paradigm to study whether the temporal structure of inputs influences the synaptic transmission onto pontine neurons. Pairs of electrical stimuli applied to the cerebral peduncle resulted in a marked enhancement of the amplitude of the second EPSP for interstimulus intervals of 10 100 ms. Delays >200 ms left the EPSP amplitude unaltered. These data provide evidence for a complex synaptic integration and an intrinsic connectivity within the PN too elaborate to support the previous notion that the PN are simply a relay station. PMID- 9405549 TI - Circuitry underlying antiopioid actions of orphanin FQ in the rostral ventromedial medulla. AB - Several laboratories recently identified a 17 amino-acid peptide, termed "nociceptin" or "orphanin FQ (OFQ)", as the endogenous ligand for the LC132 (or "opioid receptor-like1") receptor. Taken together with the fact that the cellular effects of OFQ are to a large extent opioid-like, the close relationship between the LC132 receptor and known opioid receptors raised expectations that the behavioral effects of this peptide would resemble those of opioids. However studies of the role of OFQ in nociception have not provided a unified view. The aim of the present study was to use a combination of electrophysiological and pharmacological techniques to characterize the actions of OFQ in a brain region in which the circuitry mediating the analgesic actions of opioids has been relatively well characterized, the rostral ventromedial medulla (RVM). Single cell recording was combined with opioid administration and local infusion of OFQ in the RVM of rats lightlyanesthetized with barbiturates. The tail flick reflex was used as a behavioral index of nociceptive responsiveness. Two classes of physiologically identifiable RVM neurons with distinct responses to opioids have been characterized. -cells are activated, although indirectly, by opioids, and there is strong evidence that this activation is crucial to opioid antinociception. -cells, thought to enable nociception, are directly inhibited by opioids. Cells of a third class, cells, do not respond to opioids and whether or not they have any role in nociceptive modulation remains an open question. OFQ infused within the RVM profoundly suppressed the firing of all classes of RVM neurons, blocking opioid-induced activation of -cells. The antinociceptive effects of a micro-opioid agonist infused at the same site were significantly attenuated in these animals. Those of systemically administered morphine, which can produce its antinociceptive effects by acting at a number of CNS sites, were not blocked by RVM OFQ. Inasmuch as activation of -cells can account for the antinociceptive action of opioids within the RVM, these results demonstrate that, at least within the medulla, OFQ can exert a functional "antiopioid" effect by suppressing firing of this cell class. However to the extent that antinociceptive and pronociceptive outflows from various brain regions involved in both transmission and modulation of nociception are active under different conditions, focal application of OFQ in different regions could potentially produce either hypalgesia or hyperalgesia. PMID- 9405550 TI - Regulation of N- and L-type Ca2+ channels in adult frog sympathetic ganglion B cells by nerve growth factor in vitro and in vivo. AB - To examine mechanisms responsible for the long-term regulation of Ca2+-channels in an adult neuron, changes in whole cell Ba2+ current (IBa) were examined in adult bullfrog sympathetic ganglion B cells in vitro. Cells were cultured at low density in defined, serum free medium. After 15 days, total IBa was similar to the initial value, whereas IBa density was reduced by approximately 36%, presumably due to an increase in neuronal surface area. By contrast, IBa density remained constant after 6-15 days in the presence of murine beta-NGF (200 ng/ml), and total IBa was almost doubled. Inclusion of cytosine arabinoside (Ara-C; 10 microM) to inhibit proliferation of nonneuronal cells, did not affect the survival of neurons in the absence of nerve growth factor (NGF) nor did it attenuate IBa. Ara-C did not prevent the effect of NGF on IBa. There were three independent components to the action of NGF; during 6-9 days, it increased omega conotoxin-GVIA-sensitive N-type IBa (IBa,N); increased nifedipine-sensitive L type IBa (IBa,L) and decreased inactivation of the total Ba2+ conductance (gBa). The latter effect involved a selective decrease in the amplitude of one of the four kinetic components that describe the inactivation process. Total IBa was also 55.8% larger than control in the somata of B cells acutely dissociated from leopard frogs that had received prior subcutaneous injections of NGF. By contrast, injection of NGF antiserum decreased total IBa by 29.4%. There was less inactivation of gBa in B cells from NGF-injected animals than in cells from animals injected with NGF antiserum (P < 0.001). These data suggest that NGF-like molecule(s) play(s) a role in the maintenance of IBa in an adult amphibian sympathetic neuron; the presence of NGF may allow the neuron to maintain a constant relationship between cell size and current density. They also show that IBa inactivation in an adult neuron can be modulated in a physiologically relevant way by an extracellular ligand. PMID- 9405551 TI - Compartmental model of vertebrate motoneurons for Ca2+-dependent spiking and plateau potentials under pharmacological treatment. AB - In contrast to the limited response properties observed under normal experimental conditions, spinal motoneurons generate complex firing patterns, such as Ca2+ dependent regenerative spiking and plateaus, in the presence of certain neurotransmitters and ion-channel blockers. We have developed a quantitative motoneuron model, based on turtle motoneuron data, toinvestigate the roles of specific ionic currents and the effects of their soma and dendritic distribution in generating these complex firing patterns. In addition, the model is used to explore the effects of multiple ion channel blockers and neurotransmitters that are known to modulate motoneuron firing patterns. To represent the distribution of ionic currents across the soma and dendrites, the model contains two compartments. The soma compartment, representing the soma and proximal dendrites, contains Hodgkin-Huxley-like sodium (INa) and delayed rectifier K+ (IK-dr) currents, an N-like Ca2+ current (ICa-N), and a calcium-dependent K+ current [IK(Ca)]. The dendritic compartment, representing the lumped distal dendrites, contains, in addition to ICa-N and IK(Ca) as in the soma, a persistent L-like calcium current (ICa-L). We determined kinetic parameters for INa, IK-dr, ICa-N, and IK(Ca) in order to reproduce normal action-potential firing observed in turtle spinal motoneurons, including fast and slow afterhyperpolarizations (AHPs) and a linear steady-state frequency-current relation. With this parameter set as default, a sequence of pharmacological manipulations were systematically simulated. A small reduction of IK-dr [mimicking the experimental effect of tetraethylammonium (TEA) in low concentration] enhanced the slow AHP and caused calcium spiking (mediated by ICa-N) when INa was blocked. Firing patterns observed experimentally in high TEA [and tetrodotoxin (TTX)], namely calcium spikes riding on a calcium plateau, were reproduced only when both IK-dr and IK(Ca) were reduced. Dendritic plateau potentials, mediated by ICa-L, were reliably unmasked when IK(Ca) was reduced, mimicking the experimental effect of the bee venom apamin. The effect of 5-HT, which experimentally induces the ability to generate calcium-dependent plateau potentials but not calcium spiking, was reproduced in the model by reducing IK(Ca) alone. The plateau threshold current level, however, was reduced substantially if a simultaneous increase in ICa-L was simulated, suggesting that serotonin (5-HT) induces plateau potentials by regulating more than one conductance. The onset of the plateau potential showed significant delays in response to near-threshold, depolarizing current steps. In addition, the delay times were sensitive to the current step amplitude. The delay and its sensitivity were explained by examining the model's behavior near the threshold for plateau onset. This modeling study thus accurately accounts for the basic firing behavior of vertebrate motoneurons as well as a range of complex firing patterns invoked by ion-channel blockers and 5-HT. In addition, our computational results support the hypothesis that the electroresponsiveness of motoneurons depends on a nonuniform distribution of ionic conductances, and they predict modulatory effects of 5-HT and properties of plateau activation that have yet to be tested experimentally. PMID- 9405552 TI - Behavioral function of glutamatergic interneurons in the feeding system of Lymnaea: plateauing properties and synaptic connections with motor neurons. AB - Intracellular recording techniques were used to examine the electrical properties and behavioral function of a novel type of retraction phase interneuron, the N2 ventral (N2v) cells in the feeding network of the snail Lymnaea. The N2vs were compared with the previously identified N2 cells that now are renamed the N2 dorsal (N2d) cells. The N2vs are a bilaterally symmetrical pair of electrotonically coupled plateauing interneurons that are located on the ventral surfaces of the buccal ganglia. Their main axons project to the opposite buccal ganglion, but they have an additional fine process in the postbuccal nerve. N2v plateaus that outlast the duration of the stimulus can be triggered by depolarizing current pulses and prematurely terminated by applied hyperpolarizing pulses. Gradually increasing the amplitude of depolarizing pulses reveals a clear threshold for plateau initiation. N2v plateauing persists in a high Mg2+/nominally zero Ca2+ saline that blocks chemical synaptic connections, suggesting an endogenous mechanism for plateau generation. The N2vs fire sustained bursts of action potentials throughout the N2/rasp phase of the fictive feeding cycle and control the retraction phase feeding motor neurons. The N2vs excite the B3 and B9 feeding motor neurons to fire during the rasp phase of the feeding cycle. They also inhibit the B7 and B8 feeding motor neurons. The B8 cells recover from inhibition and fire during the following swallowing phase. These synaptic connections appear to be monosynaptic as they persist in high Mg2+/high Ca2+ (HiDi) saline that blocks polysynaptic pathways. Strong current induced plateaus in the N2vs generate brief inhibitory postsynaptic responses in the B4CL rasp phase motor neurons, but this was due to the indirect N2v --> N2d - > B4CL pathway. The N2vs are coupled electrotonically to the N2d cells, and triggering plateau in a N2v usually induced one or two spikes in a N2d. Previous experiments showed that the N2ds generate plateau potentials during a fictive feeding cycle. Here we show that the main component of the "plateauing" waveform is due to the electrotonic coupling with the N2v cells. The differential synaptic connections of the N2v and N2d cells with retraction phase motor neurons results in a sequence of motor neuron burst activity B9 --> B4CL --> B8 that produces the full retraction (rasp --> swallow) movements of the feeding apparatus (buccal mass). We conclude that the N2v cells are an essential component of the interneuronal network required to produce feeding motor neuron activity. PMID- 9405553 TI - Glutamatergic N2v cells are central pattern generator interneurons of the lymnaea feeding system: new model for rhythm generation. AB - We aimed to show that the paired N2v (N2 ventral) plateauing cells of the buccal ganglia are important central pattern generator (CPG) interneurons of the Lymnaea feeding system. N2v plateauing is phase-locked to the rest of the CPG network in a slow oscillator (SO)-driven fictive feeding rhythm. The phase of the rhythm is reset by artificially evoked N2v bursts, a characteristic of CPG neurons. N2v cells have extensive input and output synaptic connections with the rest of the CPG network and the modulatory SO cell and cerebral giant cells (CGCs). Synaptic input from the protraction phase interneurons N1M (excitatory), N1L (inhibitory), and SO (inhibitory-excitatory) are likely to contribute to a ramp-shaped prepotential that triggers the N2v plateau. The prepotential has a highly complex waveform due to progressive changes in the amplitude of the component synaptic potentials. Most significant is the facilitation of the excitatory component of the SO --> N2v monosynaptic connection. None of the other CPG interneurons has the appropriate input synaptic connections to terminate the N2v plateaus. The modulatory function of acetylcholine (ACh), the transmitter of the SO and N1M/N1Ls, was examined. Focal application of ACh (50-ms pulses) onto the N2v cells reproduced the SO --> N2v biphasic synaptic response but also induced long term plateauing (20-60 s). N2d cells show no endogenous ability to plateau, but this can be induced by focal applications of ACh. The N2v cells inhibit the N3 tonic (N3t) but not the N3 phasic (N3p) CPG interneurons. The N2v --> N3t inhibitory synaptic connection is important in timing N3t activity. The N3t cells recover from this inhibition and fire during the swallow phase of the feeding pattern. Feedback N2v inhibition to the SO, N1L protraction phase interneurons prevents them firing during the retraction phase of the feeding cycle. The N2v - > N1M synaptic connection was weak and only found in 50% of preparations. A weak N2v --> CGC inhibitory connection prevents the CGCs firing during the rasp (N2) phase of the feeding cycle. These data allowed a new model for the Lymnaea feeding CPG to be proposed. This emphasizes that each of the six types of CPG interneuron has a unique set of synaptic connections, all of which contribute to the generation of a full CPG pattern. PMID- 9405554 TI - Glutamate is the transmitter for N2v retraction phase interneurons of the Lymnaea feeding system. AB - Electrophysiological and pharmacological methods were used to examine the role of glutamate in mediating the excitatory and inhibitory responses produced by the N2v rasp phase neurons on postsynaptic cells of the Lymnaea feeding network. The N2v --> B3 motor neuron excitatory synaptic response could be mimicked by focal or bath application of -glutamate at concentrations of >/=10(-3) M. Quisqualate and alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) were potent agonists for the B3 excitatory glutamate receptor (10(-3) M), whereas kainate only produced very weak responses at the same concentration. This suggested that non-N-methyl--aspartate (NMDA), AMPA/quisqualate receptors were present on the B3 cell. The specific non-NMDA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3 dione (CNQX; 10(-5) M) blocked 85% of the excitatory effects on the B3 cell produced by focal application of glutamate (10(-3) M), confirming the presence of non-NMDA receptors. CNQX also blocked the major part of the excitatory postsynaptic potentials on the B3 cell produced by spontaneous or current-evoked bursts of spikes in the N2v cell. As with focal application of glutamate, a small delayed component remained that was CNQX insensitive. This provided direct evidence that glutamate acting via receptors of the non-NMDA, AMPA/quisqualate type were responsible for mediating the main N2v --> B3 cell excitatory response. NMDA at 10(-2) M also excited the B3 cell, but the effects were much more variable in size and absent in one-third of the 25 B3 cells tested. NMDA effects on B3 cells were not enhanced by bath application of glycine at 10(-4) M or reduction of Mg2+ concentration in the saline to zero, suggesting the absence of typical NMDA receptors. The variability of the B3 cell responses to NMDA suggested these receptors were unlikely to be the main receptor type involved with N2v --> B3 excitation. Quisqualate and AMPA at 10(-3) M also mimicked N2v inhibitory effects on the B7 and B8 feeding motor neurons and the modulatory slow oscillator (SO) interneuron, providing further evidence for the role of AMPA/quisqualate receptors. Similar effects were seen with glutamate at the same concentration. However, CNQX could not block either glutamate or N2v inhibitory postsynaptic responses on the B7, B8, or SO cells, suggesting a different glutamate receptor subtype for inhibitory responses compared with those responsible for N2v --> B3 excitation. We conclude that glutamate is a strong candidate transmitter for the N2v cells and that AMPA/quisquate receptors of different subtypes are likely to be responsible for the excitatory and inhibitory postsynaptic responses. PMID- 9405555 TI - Behavior-dependent activities of a central pattern generator in freely behaving Lymnaea stagnalis. AB - Cyclic or repeated movements are thought to be driven by networks of neurons (central pattern generators) that are dynamic in their connectivity. During two unrelated behaviors (feeding and egg laying), we investigated the behavioral output of the buccal pattern generator as well as the electrical activity of a pair of identified interneurons that have been shown to be involved in setting the level of activity of this pattern generator (PG). Analysis of the quantile plots of the parameters that describe the behavior (movements of the buccal mass) reveals that during egg laying, the behavioral output of the PG is different compared with that during feeding. Comparison of the average durations of the different parts of the buccal movements showed that during egg laying, the duration of one specific part of buccal movement is increased. Correlated with these changes in the behavioral output of the PG were changes in the firing rate of the cerebral giant neurons (CGC), a pair of interneurons that have been shown to modulate the activity of the PG by means of multiple synaptic contacts with neurons in the buccal ganglion. Interval- and autocorrelation histograms of the behavioral output and CGC spiking show that both the PG output and the spiking properties of the CGCs are different when comparing egg-laying animals with feeding animals. Analysis of the timing relations between the CGCs and the behavioral output of the PG showed that both during feeding and egg laying, the electrical activity of the CGCs is largely in phase with the PG output, although small changes occur. We discuss how these results lead to specific predictions about the kinds of changes that are likely to occur when the animal switches the PG from feeding to egg laying and how the hormones that cause egg laying are likely to be involved. PMID- 9405556 TI - Modulation of multiple potassium currents by metabotropic glutamate receptors in neurons of the hypothalamic supraoptic nucleus. AB - We studied the effects of activation of the metabotropic glutamate receptors on intrinsic currents of magnocellular n urons of the supraoptic nucleus (SON) with whole cell patch-clamp and conventional intracellular recordings in coronal slices (400 micron) of the rat hypothalamus. Trans-(+/-)-1-amino-1,3-cyclopentane dicarboxylic acid (trans-ACPD, 10-100 microM), a broad-spectrum metabotropic glutamate receptor agonist, evoked an inward current (18.7 +/- 3.45 pA) or a slow depolarization (7.35 +/- 4.73 mV) and a 10-30% decrease in whole cell conductance in approximately 50% of the magnocellular neurons recorded at resting membrane potential. The decrease in conductance and the inward current were caused largely by the attenuation of a resting potassium conductance because they were reduced by the replacement of intracellular potassium with an equimolar concentration of cesium or by the addition of potassium channel blockers to the extracellular medium. In some cells, trans-ACPD still elicited a small inward current after blockade of potassium currents, which was abolished by the calcium channel blocker, CdCl2. Trans-ACPD also reduced voltage-gated and Ca2+-activated K+ currents in these cells. Trans-ACPD reduced the transient outward current (IA) by 20-70% and/or the IA-mediated delay to spike generation in approximately 60% of magnocellular neurons tested. The cells that showed a reduction of IA generally also showed a 20-60% reduction in a voltage-gated, sustained outward current. Finally, trans-ACPD attenuated the Ca2+-dependent outward current responsible for the afterhyperpolarization (IAHP) in approximately 60% of cells tested. This often revealed an underlying inward current thought to be responsible for the depolarizing afterpotential seen in some magnocellular neurons. (RS)-3,5 dihydroxyphenylglycine, a group I receptor-selective agonist, mimicked the effects of trans-ACPD on the resting and voltage-gated K+ currents. (RS)-alpha methyl-4-carboxyphenylglycine, a group I/II metabotropic glutamate receptor antagonist, blocked these effects. A group II receptor agonist, 2S,1'S,2'S 2carboxycyclopropylglycine and a group III receptor agonist, (+)-2-amino-4 phosphonobutyric acid, had no effect on the resting or voltage-gated K+ currents, indicating that the reduction of K+ currents was mediated by group I receptors. About 80% of the SON cells that were labeled immunohistochemically for vasopressin responded to metabotropic glutamate receptor activation, whereas only 33% of labeled oxytocin cells responded, suggesting that metabotropic receptors are expressed preferentially in vasopressinergic neurons. These data indicate that activation of the group I metabotropic glutamate receptors leads to an increase in the postsynaptic excitability of magnocellular neurons by blocking resting K+ currents as well as by reducing voltage-gated and Ca2+-activated K+ currents. PMID- 9405557 TI - Receptive field changes after strokelike cortical ablation: a role for activation dynamics. AB - The reorganization of neural activity that takes place after stroke is of paramount importance in producing functional recovery. Experimental stroke models have suggested that this reorganization may have two phases, but physiology alone cannot fully resolve what causes each phase. Computer modeling suggests that these phases might involve an initial change in dynamics occurring immediately, followed by synaptic plasticity. We combined physiological recording from macaque middle temporal cortex (area MT) with a neural network computer model to examine this first phase of altered cortical function after a small, experimentally induced cortical lesion. Major receptive field (RF) changes seen in the first few days postlesion included both expansion and contraction of receptive fields. Although only expansion could be reproduced in an initial model, addition of inhibitory interneuron loss in a ring around the primary ablation, suggested by immunohistochemical examination, permitted contraction to be replicated as well. We therefore predict that this immunochemical observation reflects an immediate extension of the lesion rather than a late response. Additionally our model successfully predicted a correlation between increased firing rate and RF size. Our model suggests that activation dynamics alone, without anatomic remodeling, can cause the large receptive field changes that allow the rapid behavioral recovery seen after middle temporal lesions. PMID- 9405558 TI - Frequency-dependent information flow from the entorhinal cortex to the hippocampus. AB - Storage and retrieval of information in the hippocampus is dependent on information transfer from the entorhinal cortex (EC). We studied how the separate pathways from layer II and III of the EC to the hippocampus are selected for information transfer during repetitive synaptic stimulation. Intracellular recordings were made from EC layer II and III projection cells in horizontal combined EC-hippocampal slices. Synaptic responses to stimulation of deep layers or the lateral EC with stimulus intensities approximately 70% of that required to elicit an action potential were analyzed during short trains of repetitive stimulation. The threshold intensities for induction of action potentials were in layer II cells 8.2 +/- 3.8 (SE) V, significantly larger than 4.4 +/- 1.5 V in type 1, and 5.2 +/- 3.3 V in type 2 layer III cells, respectively. During repetitive subthreshold stimulation with frequencies below 5 Hz the pathway from the EC layer II remained quiet and was preferentially activated with stimulation frequencies above 5 Hz. In contrast the EC layer III cells responded preferentially to low stimulus frequencies (<10 Hz) and became strongly inhibited when synaptically stimulated with frequencies above 10 Hz. Interestingly during stimulus frequencies between 5 and 10 Hz the likelihood that both layer II and III cells fire was large. Thus a frequency switch operates in the entrohinal cortex regulating output of layer II and III cells to the hippocampus. We suggest that such frequency dependent regulation of information flow presents a new principle of neuronal information processing. PMID- 9405559 TI - Dopaminergic modulation of inhibitory glutamate receptors in the lobster stomatogastric ganglion. AB - The intrinsic rhythmicity of the spiny lobster stomatogastric ganglion (STG) is strongly influenced by the strengths of the graded synapses between identified cells within the neural network. These synaptic strengths can be powerfully influenced by chemical neuromodulators such as dopamine and serotonin. Most of the intraganglionic chemical synapses in the STG are mediated by postsynaptic inhibitory glutamate receptors (IGluRs). To determine whether or not direct effects on these IGluRs contribute to the modulation of synaptic strength, unidentified STG neurons were extracted into primary culture and the effects of these aminergic neuromodulators on the glutamate-evoked membrane current were assessed. Dopamine (100 microM) reliably and significantly reduced the whole cell slope conductance of all IGluRs tested. Serotonin (20 microM) never affected the IGlu response, although it clearly altered other cellular membrane properties. Although all identified STG neurons may not conform to these observations, the data reveal a specific dopamine-activated modulatory pathway within cultured neurons that reduces IGluR slope conductance. The relationship between IGluR modulation and net synaptic modulation in situ contributes to an emerging model in which synaptic strengths can be multiply modulated at different functional sites, yielding a complex, distributed, and state-dependent regulatory structure. PMID- 9405560 TI - On the detection and measurement of synchrony in neural populations by coherence analysis. AB - This study considers the possibility of using coherence analysis for detection and measurement of synchrony (correlations) in large neural populations, applied to activities that are relatively easy to record in parallel. Mathematical analysis and computer simulations are used to examine the behavior of the coherence function between both unitary and population-aggregate activity (UTA coherence) and the aggregate activities of two populations (ATA coherence). The results indicate that for a large population showing partial correlations, the UTA coherence function is almost zero at all frequencies for the uncorrelated units. However, unless the synchrony is very restricted, its value is nonzero (i.e., statistically significant by common criteria) at each frequency of synchrony for the units that show correlations to other units. Moreover, this value is indicative of the strength of synchrony for any given unit. These properties enable the identification of the correlated units in a sample of unit/population activities simultaneously recorded in a series of experiments, and hence the detection of synchrony. The extent of synchrony can then be estimated as the fraction of such units in the sample, whereas the values of the UTA coherences in the sample can be used to estimate the strength and its distribution within the population. Similarly, the ATA coherence function is generally nonzero (significant) at the frequencies where there are correlations between members of two large populations. This enables the easy detection of such correlations from simultaneously recorded population activities. However, this function is a very sensitive index of synchrony and even shows saturation effects. It may therefore be used as a general measure of synchrony only under restricted conditions. PMID- 9405561 TI - Time-dependent changes in excitability after one-trial conditioning of Hermissenda. AB - The visual system of Hermissenda has been studied extensively as a site of cellular plasticity produced by classical conditioning. A one-trial conditioning procedure consisting of light paired with the application of serotonin (5-HT) to the exposed, but otherwise intact, nervous system produces suppression of phototactic behavior tested 24 h after conditioning. Short- and long-term enhancement (STE and LTE) of excitability in identified type B photoreceptors is a cellular correlate of one-trial conditioning. LTE can be expressed in the absence of STE suggesting that STE and LTE may be parallel processes. To examine the development of enhancement, we studied its time-dependent alterations after one-trial conditioning. Intracellular recordings from identified type B photoreceptors of independent groups collected at different times after conditioning revealed that enhanced excitability follows a biphasic pattern in its development. The analysis of spikes elicited by 2 and 30 s extrinsic current pulses at different levels of depolarization showed that enhancement reached a peak 3 h after conditioning. From its peak, excitability decreased toward baseline control levels 5-6 h after conditioning followed by an increase to a stable plateau at 16 to 24 h postconditioning. Excitability changes measured in cells from unpaired control groups showed maximal changes 1 h posttreatment that rapidly decremented within 2 h. The conditioned stimulus (CS) elicited significantly more spikes 24 h postconditioning for the conditioned group as compared with the unpaired control group. The analysis of the time-dependent development of enhancement may reveal the processes underlying different stages of memory for this associative experience. PMID- 9405563 TI - Contribution of auditory cortex to acoustical orientation in cats under conditions of discordant auditory reafference. AB - Head-orienting responses (ORs) evoked by a stationary source of sound typically terminate in small undershoots in normal hearing cats or in large undershoots (hypometria) if the auditory cortex is ablated bilaterally. In the present study, ORs executed by cats were studied using a procedure in which the OR produced an isogonal rotation of the sound source, i.e., response-produced change (reafference) in the acoustic stimulus was distorted. Under this condition (discordant auditory reafference), ORs terminated in large overshoots (hypermetria) in the normal hearing cats. This result indicates that experimental distortion of response-produced auditory feedback resulted in an "on-line" modification of ORs by the normal hearing cats. In the cats with auditory cortex ablated, ORs terminated in large undershoots (hypometria), suggesting that auditory cortex is a necessary component of the central auditory system for processing reafferent acoustic stimuli that normally occur during head rotation in a sound field. PMID- 9405562 TI - Imaging of calcium in Drosophila larval motor nerve terminals. AB - Calcium measurements in the presynaptic terminal are essential in the investigation of mechanisms underlying neurotransmitter release. To enhance the genetic analysis of secretory mechanisms, we have developed Ca2+ imaging techniques for Drosophila larval motor nerve terminals. We studied Ca2+ signals in "big" (type Ib) and "small" (type Is) boutons that innervate ventral longitudinal muscles 6 and 7 in each abdominal segment of Canton-S (CS)-strain 3rd instar larvae. The indicator fluo-3 in conjunction with confocal microscopy was used to detect stimulus-dependent changes in [Ca2+]i. The Ca2+ signals were reliable and reproducible, and the resting fluorescence remained constant throughout the experiments. The Ca2+ signals increased with stimulus frequency from 5 to 20 Hz for both bouton types. No significant differences in the Ca2+ signals were seen between the two bouton types at 5 and 20 Hz, but there was a difference at 10 Hz. The decay of the Ca2+ signal was more prolonged after 20-Hz stimulation than after 5 and 10 Hz. At the single-synapse level, the secretory efficacy of Is synapses is greater than that of Ib synapses, but our data show that factors other than differences in Ca2+ entry may govern the strength of synaptic transmission. PMID- 9405564 TI - Loss of long-lasting potentiation mediated by group III mGluRs in amygdala neurons in kindling-induced epileptogenesis. AB - Long-lasting modifications of synaptic transmission can be induced in the amygdala by electrical stimulation as done in the long-term potentiation (LTP) model of learning and memory and the kindling model of epilepsy. The present study reports for the first time a long-lasting potentiation (LLP) of synaptic transmission that is induced pharmacologically by the activation of group III metabotropic glutamate receptors (mGluRs) in basolateral amygdala (BLA) neurons. In whole cell voltage-clamp mode, BLA neurons were recorded in brain slices from control rats and rats with amygdala-kindled seizures. The group III mGluR agonist -2-amino-4-phosphonobutyrate (-AP4, 10 microM) induced LLP of monosynaptic excitatory postsynaptic currents (EPSCs) evoked by electrical stimulation in the lateral amygdala (maximum 258 +/- 50% of predrug control; means +/- SE) in control (n = 7) but not in kindled neurons(n = 6). LLP was measured 15 min after the superfusion of -AP4, lasted for >45 min, and was not accompanied by postsynaptic membrane changes. -AP4 induced LLP was prevented by the group III mGluR antagonist (S)-2-methyl-2-amino-4-phosphonobutyrate (MAP4; 100 microM, n = 6) but not the group II mGluR antagonist (2S, 3S,4S)-2-methyl-2 carboxycyclopropylglycine (MCCG; 100 microM, n = 3). LLP was not observed after superfusion of the group II mGluR agonist (2S,3S,4S)-2 (carboxycyclopropyl)glycine (-CCG; 1.0 and 10 microM) in either control (n = 13) or kindled (n = 10) neurons. If the underlying mechanisms and the functional significance of pharmacologically induced LLP are similar to those of LTP, the loss of -AP4 induced LLP in kindled neurons may be a neurobiological correlate of learning and memory deficits in kindled animals and long-term alterations of brain functions in patients with epilepsies. PMID- 9405565 TI - Right-left interactions between rostral scratch networks generate rhythmicity in the preenlargement spinal cord of the turtle. AB - We examined the rhythmogenic capacity of the midbody D3-D7 spinal cord during stimulation of the rostral scratch reflex in turtles. Fictive scratching was recorded bilaterally as electroneurograms (ENGs) from prehindlimb enlargement nerves [transverse D7 (TD7) and oblique D7 (OD7)] and hip flexor nerves (HF). TD7 and OD7 innervate transverse- and oblique-abdominus muscles, respectively. D3-end preparations had intact spinal cords caudal to a D2-D3 transection site. Unilateral stimulation of the rostral receptive field in D3-end preparations evoked rhythmic bursting in the ipsilateral (ipsi) HF nerve and bilateral rhythmic discharge in the TD7 and OD7 nerves. Right HF bursts were coactive with right TD7 and left OD7 bursts and alternated with left TD7 and right OD7 bursts. D3-D7 preparations received a second spinal transection at the caudal end of segment D7, thus resulting in activation of strictly preenlargement circuitry in response to rostral scratch stimulation and preventing activation of hindlimb enlargement circuitry in segments D8-S2. D3-D7 preparations responded to unilateral stimulation with modulated or tonic discharge in the ipsi TD7 and contralateral (contra) OD7 nerves. In contrast, bilateral stimulation reestablished robust bursting in which coactive right TD7-left OD7 bursts alternated with coactive left TD7-right OD7 bursts. These data imply that TD7 circuit modules make 1) crossed excitatory connections with contra OD7 circuitry, 2) crossed inhibitory connections with contra TD7 circuitry, and 3) uncrossed inhibitory connections with ipsi OD7 circuitry. Our results also suggest that bilateral stimulation evokes rhythmic alternation in the preenlargment cord by simultaneously exciting reciprocally inhibitory circuit modules. PMID- 9405566 TI - Inhibition of dendritic calcium influx by activation of G-protein-coupled receptors in the hippocampus. AB - Gi proteins inhibit voltage-gated calcium channels and activate inwardly rectifying K+ channels in hippocampal pyramidal neurons. The effect of activation of G-protein-coupled receptors on action potential-evoked calcium influx was examined in pyramidal neuron dendrites with optical and extracellular voltage recording. We tested the hypotheses that 1) activation of these receptors would inhibit calcium channels in dendrites; 2) hyperpolarization resulting from K+ channel activation would deinactivate low-threshold, T-type calcium channels on dendrites, increasing calcium influx mediated by these channels; and 3) activation of these receptors would inhibit propagation of action potentials into dendrites, and thus indirectly decrease calcium influx. Activation of adenosine receptors, which couple to Gi proteins, inhibited calcium influx in cell bodies and proximal dendrites without inhibiting action-potential propagation into the proximal dendrites. Inhibition of dendritic calcium influx was not changed in the presence of 50 microM nickel, which preferentially blocks T-type channels, suggesting influx through these channels is not increased by activation of G proteins. Adenosine inhibited propagation of action potentials into the distal branches of pyramidal neuron dendrites, leading to a three- to fourfold greater inhibition of calcium influx in the distal dendrites than in the soma or proximal dendrites. These results suggest that voltage-gated calcium channels are inhibited in pyramidal neuron dendrites, as they are in cell bodies and terminals and thatG-protein-mediated inhibition of action-potential propagation can contribute substantially to inhibition of dendritic calcium influx. PMID- 9405567 TI - Corticofugal amplification of subcortical responses to single tone stimuli in the mustached bat. AB - Since 1962, physiological data of corticofugal effects on subcortical auditory neurons have been controversial: inhibitory, excitatory, or both. An inhibitory effect has been much more frequently observed than an excitatory effect. Recent studies performed with an improved experimental design indicate that corticofugal system mediates a highly focused positive feedback to physiologically "matched" subcortical neurons, and widespread lateral inhibition to "unmatched" subcortical neurons, in order to adjust and improve information processing. These results lead to a question: what happens to subcortical auditory responses when the corticofugal system, including matched and unmatched cortical neurons, is functionally eliminated? We temporarily inactivated both matched and unmatched neurons in the primary auditory cortex of the mustached bat with muscimol (an agonist of inhibitory synaptic transmitter) and measured the effect of cortical inactivation on subcortical auditory responses. Cortical inactivation reduced auditory responses in the medial geniculate body and the inferior colliculus. This reduction was larger (60 vs. 34%) and faster (11 vs. 31 min) for thalamic neurons than for collicular neurons. Our data indicate that the corticofugal system amplifies collicular auditory responses by 1.5 times and thalamic responses by 2.5 times on average. The data are consistant with a scheme in which positive feedback from the auditory cortex is modulated by inhibition that may mostly take place in the cortex. PMID- 9405568 TI - Monkey posterior parietal cortex neurons antidromically activated from superior colliculus. AB - The connection between the posterior parietal cortex (PPC) and the superior colliculus (SC) was investigated by antidromically activating neurons within the lateral intraparietal (LIP) area with single-pulse stimulation delivered to the intermediate layers of the SC. To dissociate visual and saccade-related responses, the discharge properties of the identified efferent neurons were studied in the delayed visually guided saccade task and the memory guided saccade task. We found that the great majority (74%) of the identified LIP efferent neurons have a peripheral visual receptive field, typically with a broad spatial tuning. About two-thirds (64%) exhibited sustained activity during the delay period of the behavioral tasks, during which the monkeys had to withhold eye movements, and 80% of these increased their activity just before the onset of saccades. Both delay and presaccadic discharges in the delayed visually guided saccade task were higher than in the memory guided saccade task. These results establish that the neuronal signal sent by LIP to the SC carries both visual and saccade-related information. PMID- 9405569 TI - Muscle fibers in regenerating crayfish motor nerves. AB - Single discrete muscle fibers were found in regenerating motor nerves in adult crayfish. The regenerating nerves were from native or transplanted ganglia in the third abdominal segments and consisted of several motor axons. The proximal end of these motor axons showed numerous sprouts. Muscle fibers in these regenerating nerves appeared newly developed and were innervated by excitatory nerve terminals. A likely source of these novel muscle fibers may be blood cells in the nerve or satellite cells from neighboring muscle. Contacts made by axon sprouts with other axon sprouts, glia, and muscle fiber, in the form of a dense bar with clustered clear vesicles, characterized the regenerating nerve. These contacts may provide a possible signaling pathway for axon regeneration and myogenesis. PMID- 9405570 TI - Local field potential oscillations in primate cerebellar cortex during voluntary movement. AB - Sustained oscillations of 13-18 Hz were observed in local field potentials (LFPs) in the cerebellar cortex of a behaving monkey. These oscillations, which appeared to be generated in the granular cell layer, were particularly prominent in the paramedian lobule. The oscillatory activity decreased during drowsiness or extreme arousal and occurred most often when the animal was immobile but alert. In a task requiring the animal to move the arm approximately 1 s after an auditory cue, the oscillations stopped some 150-200 ms after the cue, resumed 200 300 ms later, and stopped again 50-100 ms before movement onset. This modulation pattern was observed with consistency only when the animal responded reliably to the auditory cue. The results suggest that the cerebellum could be involved in some higher level of integration particularly during complex sensorimotor behavior. PMID- 9405571 TI - Bidirectional electrical coupling between inspiratory motoneurons in the newborn mouse nucleus ambiguus. AB - Some spinal and brain stem motoneurons are electrically coupled in the early postnatal period. To test whether respiratory motoneurons in the brain stem are electrically coupled, we performed single and dual whole cell patch recordings from presumptive motoneurons in the nucleus ambiguus in a rhythmically active brain stem slice from newborn mice. Two of eight (25%) biocytin-injected neurons showed dye-coupling and 4 of 11 (36%) of intracellularly recorded pairs of neurons showed evidence of bidirectional electrical coupling. Impulse activity in one cell elicited small spikelets in the other and hyperpolarization of one cell led to hyperpolarization of the other with a coupling ratio (DeltaV2:DeltaV1) of 0.03-0.14. We conclude that inspiratory ambiguus motoneurons in the newborn mouse brain stem are bidirectionally electrically coupled, which may serve to transmit or coordinate signals, chemical or electrical. PMID- 9405572 TI - George M. Briggs (1919-1989). PMID- 9405573 TI - Anthony August Albanese (1908-1994). PMID- 9405574 TI - Dietary copper in the physiology of the microcirculation. AB - Dietary copper has long been known to be essential for cardiovascular homeostasis. However, the role of copper and cuproenzymes in the normal control of vascular physiology is not well understood. Most studies in the cardiovascular system have focused on copper deficiency-induced defects in the heart or large vessels. Recently, attention has also focused on the effects of copper deficiency in the microcirculation or the small blood vessels that control blood flow, nutrient and waste exchange, and peripheral vascular resistance. Studies in the microcirculation demonstrate that copper is important in mechanisms of macromolecular leakage, platelet-endothelial interactions and vascular smooth muscle reactivity. There is a significantly greater leakage of proteins from postcapillary venules in copper-deficient rats in response to mast cell-released histamine. This response appears to be the result of increased numbers of mast cells and thereby increased available histamine. Copper deficiency also causes an inhibition of in vivo thrombogenesis, which appears to be related to an inhibition of platelet adhesion. Subsequent studies have demonstrated that this is probably caused by a diminished concentration of the adhesion molecule von Willebrand factor. Nitric oxide (NO)-mediated arteriole vasodilation is also compromised in copper-deficient rats. This functional deficit to NO can be reversed by the addition of Cu, Zn-superoxide dismutase (SOD), suggesting that degradation of NO by superoxide anion occurs during copper deprivation. These observations demonstrate that dietary copper is necessary for several microvascular control mechanisms affecting inflammation, microhemostasis and regulation of peripheral blood flow. PMID- 9405575 TI - In vivo dopamine metabolism is altered in iron-deficient anemic rats. AB - Previous studies of dopamine metabolism in iron-deficient rats demonstrated an elevation in extraneuronal levels of dopamine and a depression in the number of dopamine D2 receptors; however, the importance of anemia per se and the reversibility of these observations are not completely resolved. The purpose of this study was to determine if in vivo reuptake of caudate dopamine is altered by iron deficiency anemia, if it is reversible with iron therapy, and if anemia per se produced the same effects on dopamine metabolism. Male Sprague-Dawley rats (21 d old) were fed an iron-deficient diet (4 mg Fe/kg diet) and then iron repleted (5 mg iron dextran), or were fed an iron adequate diet (35 mg Fe/kg diet) and then given phenylhydrazine to induce hemolytic anemia. In vivo microdialysis was performed in steady-state conditions both before and after iron or no therapy and was followed by an intraperitoneal injection of a dopamine reuptake blocker (cocaine-HCl 30 mg/kg). Thirty percent higher extracellular dopamine levels in the caudate-putamen were observed in iron-deficient rats compared with control rats, but no differences were observed in tissue levels. Hemolytic anemic and iron-repleted rats had normal extracellular dopamine levels. The response to dopamine reuptake blockade was significantly attenuated in iron-deficient rats compared with control, iron-repleted, or hemolytic anemic rats. These experiments provide evidence that iron deficiency blunts the dopamine reuptake mechanism, that this is a reversible process in postweaning rats, and that anemia per se does not cause the increased extracellular dopamine levels. PMID- 9405576 TI - Unsaturated fatty acids associated with glycogen may inhibit glucose-6 phosphatase in rat liver. AB - This study was conducted to identify the nature of a glycogen-associated compound that had been shown to inhibit glucose-6 phosphatase in vitro. Glycogen was purified from the liver of fed rats by potassium hydroxyde digestion and ethanol precipitation. It inhibited glucose-6 phosphatase in microsomes isolated from rats deprived of food for 48 h. Two glycogen-associated fractions were purified by anion-exchange chromatography on DOWEX 1 (200-400 mesh). These fractions inhibited microsomal glucose-6-phosphatase activity in vitro (80 +/- 2 and 76 +/- 3% of control, respectively). After chromatography, glycogen was no longer inhibitory (101 +/- 3% of control). Because glycogen is associated with endoplasmic reticulum membranes in the liver, we tested the hypothesis that lipids could be involved in the inhibitory process. Lipids were extracted from glycogen by Folch's method and analyzed by thin-layer chromatography and gas chromatography. The glycogen-associated fractions did not contain complex lipids but contained unsaturated fatty acids, which had been shown previously to inhibit glucose-6-phosphatase in vitro. Because the concentration of unsaturated fatty acids in both fractions quantitatively accounted for the inhibition of glucose-6 phosphatase observed, and because noninhibitory chromatographed glycogen reconstituted with equivalent amounts of pure unsaturated fatty acids inhibited the enzyme as glycogen did, we conclude that unsaturated fatty acids likely constitute the glycogen-associated compound that inhibits glucose-6 phosphatase activity. PMID- 9405577 TI - Adult-onset energy restriction of rhesus monkeys attenuates oxidative stress induced cytokine expression by peripheral blood mononuclear cells. AB - We previously reported that energy restriction (ER) of mice attenuated age associated increases in serum levels of interleukin-6 (IL-6). Here, we studied peripheral blood mononuclear cells (PBMC) from male rhesus monkeys to investigate the following: 1) the production of IL-6 and other cytokines become dysregulated with aging; 2) ER influences cytokine production and mRNA expression; and, 3) oxidative stress, as induced in vitro by xanthine and xanthine oxidase (X/XOD), influences cytokine mRNA and protein levels. Two types of comparisons were made as follows: 1) between normally fed young (6-9 y) and old monkeys (22-33 y); and 2) between middle-aged monkeys (15-21 y) fed either a normal energy intake or subjected to ER (for 5.5 y at 30% less than base-line intake). IL-6 protein levels and X/XOD-induced IL-6 mRNA levels in PBMC from old monkeys were significantly greater than those in PBMC from young animals. In contrast, interleukin-1beta (IL-1beta) and interleukin-8 mRNA levels were not strongly influenced by advancing age. X/XOD, which increased levels of protein carbonyls (indicative of oxidative damage) in PBMC, induced the expression of all three cytokines. ER reduced IL-6 protein and mRNA levels induced by X/XOD and the unstimulated mRNA levels of IL-1beta. These results indicate that, in a nonhuman primate model, oxidative stress may contribute to age-associated increases in the levels of certain cytokines and that adult-onset ER partially ameliorates this alteration. PMID- 9405578 TI - Socioeconomic and demographic factors are associated with worldwide patterns of stunting and wasting of children. AB - We estimated the variability among nations in the prevalence of stunting and wasting, evaluated which national factors are associated with stunting and wasting and examined the relationship of stunting with wasting. The World Health Organization Global Database on Child Growth, a comprehensive conceptual model and a database of national factors were used with variance components and regression analyses. There was substantial variability among nations and among provinces within nations. Most national variability for stunting (76%) and wasting (66%) was explained by national factors and geographic region. Higher energy availability, female literacy and gross product were the most important factors associated with lower prevalence of stunting. The association of health expenditures and stunting differed by region. Higher immunization rate and, for Asia only, energy availability were the most important factors associated with lower prevalence of wasting. Regional differences in the relationship between stunting and wasting were accounted for by national factors. Some factors associated with stunting and wasting differ at the national level. Child malnutrition within a household is greatly influenced by issues at national and provincial levels, and intervention should be considered at all three levels. PMID- 9405579 TI - High prevalence of obesity in low income and multiethnic schoolchildren: a diet and physical activity assessment. AB - The objective of the study was to assess the prevalence of obesity and/or undernutrition and evaluate diet and activity patterns among schoolchildren from an ethnically diverse low income urban population. A cross-sectional survey of 498 children aged 9-12 y from 24 schools in low income multiethnic neighborhoods in Montreal, Canada was undertaken. Height, weight, dietary intake, physical activity record, and lifestyle and demographic characteristics were measured. There was no evidence of undernutrition because linear growth was appropriate for age, but 39.4% of children were overweight (>85th percentile NHANES II). Dietary fat intake was higher in children from single-parent families (P < 0.001) and those with mothers born in Canada. Intake of vitamins A, C, iron and folate was directly related to income sufficiency. Children who did more physical activity had significantly higher intakes of energy, calcium, iron, zinc and fiber but were not heavier. Dietary intake was systematically underreported among overweight children, i.e., their reported intakes did not meet calculated energy needs. This underreporting makes it difficult to attribute the accumulated energy imbalance to either energy intake or expenditure. PMID- 9405580 TI - Raising milk energy content retards gastric emptying of lactose in lactose intolerant humans with little effect on lactose digestion. AB - Lactose digestion improves when the energy content of a meal is raised, perhaps due to delayed gastric emptying; however, this has not been demonstrated directly. It is not known whether lactose-intolerant subjects should consume full fat or high energy milk instead of half-skimmed milk. In this study, breath 13CO2 and hydrogen (H2) measurements were combined to assess simultaneously the effect of increasing milk energy content on gastric emptying, digestion, and tolerance of lactose. On two separate days, 11 adult lactose maldigesters ingested, in the fasting state, a single dose of 710 kJ half-skimmed milk or 1970 kJ high energy milk. Both contained 18 g lactose and were supplemented with 100 mg 13C-glycine for breath 13CO2 measurement. For 6 h after milk ingestion, samples of expired breath were collected, and subjects scored their symptoms on a four-grade questionnaire. Gastric emptying was measured from excretion of breath 13CO2. The mean gastric emptying half-time was significantly longer after ingestion of high energy milk than after half-skimmed milk (84 +/- 4 vs. 64 +/- 4 min, P = 0.004). The mean area under the breath H2 excretion curve measured for 6 h was 330 +/- 61 microL/L after subjects consumed high energy milk vs. 470 +/- 82 microL/L after they consumed half-skimmed milk (P = 0.07). Mean symptom scores did not differ after ingestion of the two milks, but only two subjects experienced disturbing symptoms after high energy milk ingestion compared with five subjects after ingestion of half-skimmed milk (P = 0.56). Although ingestion of high energy milk delayed the gastric emptying of lactose for significantly longer than the ingestion of half-skimmed milk (P < 0.01), it did not lead to significant improvement in symptoms and reflected only a trend toward improved lactose digestion (P = 0.07), as measured by the area under the breath H2 excretion curve. These results indicate that it is not beneficial for most lactose intolerant subjects to replace consumption of half-skimmed milk by milk with a higher energy content. PMID- 9405581 TI - A dual-label stable-isotopic protocol is suitable for determination of folate bioavailability in humans: evaluation of urinary excretion and plasma folate kinetics of intravenous and oral doses of [13C5] and [2H2]folic acid. AB - Stable isotopic protocols for the study of folate absorption were conducted to determine the following: (1) the equivalence of the [13C5] and [2H2] forms of folic acid, and (2) the merits of short-term plasma kinetics from injected and oral doses vs. urinary excretion of [13C5] and [2H2]folates. Another objective was to evaluate the merits of protocols not involving "saturation" of subjects with nonlabeled folate. Oral administration of [13C5] and [2H2]folic acid ( approximately 500 nmol each) to adult subjects (n = 4) yielded an equivalent 24-h urinary excretion of approximately 2% of each dose (molar ratio of urinary [13C5]/[2H2]folates = 0.96 +/- 0.055; mean +/- SEM). Expression of urinary excretion as a ratio of [13C5]/[2H2]folates yielded less within-group variability than seen for absolute excretion of each form of labeled folate. In the second study, subjects received 226 nmol of [2H2]folic acid intravenously and 1010 nmol of [13C5]folic acid orally. Isotopic enrichment of plasma [2H2]folates rose rapidly and returned to near basal values by approximately 2 h postdose. In contrast, enrichment of plasma [13C5]folates was detected until 4 h after dose, whereas enrichment values were far lower than seen with [2H2]folate. Adjusting for the difference in dose, the molar response of plasma area under the curve for isotopic enrichment was 15- to 20-fold greater for injected folates. In view of this very limited short-term plasma response even with a relatively large oral dose, presumably due to hepatic first-pass uptake, these findings suggest that plasma kinetics would be of limited usefulness in assessing the relative bioavailability of nutritionally relevant oral doses of labeled folate. PMID- 9405582 TI - Dry beans inhibit azoxymethane-induced colon carcinogenesis in F344 rats. AB - Epidemiological studies show a low incidence of colon cancer in many Latin American countries where the consumption of dry beans (e.g., pinto) is high. The purpose of this study was to use rats as an animal model to obtain experimental data on the inhibition of colon carcinogenesis by dry beans. Fifty-three 5-wk-old weanling male F344 rats were randomly assigned by weight to the following groups: control (11 rats), casein diet (21 rats), and bean diet (21 rats). Animals fed the casein and bean diets were treated with the carcinogen azoxymethane (AOM) once weekly for 2 wk. Rats in the control group also consumed the casein diet but were not exposed to AOM. All diets were isocaloric. The protein concentration of the diets was adjusted to 18 g/100 g with casein, and the fat concentration was adjusted to 5 g/100 g with corn oil. Rats fed the bean diet had significantly fewer colon adenocarcinomas (P < 0.05) than rats fed the casein diet (5 vs. 22 tumors), and significantly fewer rats fed the bean diet (P < 0.05) had colonic tumors than did casein-fed rats (24 vs. 50%). Tumor multiplicity was also significantly lower for the bean-fed rats, and significantly fewer (P < 0.05) tumors per tumor-bearing rat were observed in bean-fed rats than in casein-fed rats (1.0 +/- 0.0 vs. 2.5 +/- 0.6). This study demonstrates that dry beans contain anticarcinogenic compounds capable of inhibiting AOM-induced colon cancer in rats. However, the specific anticarcinogenic components within dry beans have not been identified, and it is unclear whether dietary fiber, phytochemicals or other components within dry beans are primarily responsible for the anticarcinogenic properties of beans. PMID- 9405583 TI - A surgical model for determination of true absorption and biliary excretion of manganese in conscious swine fed commercial diets. AB - Some trace elements, such as Mn, Cu and Zn, are absorbed and quickly resecreted into the gut through the bile. When this occurs, the unabsorbed nutrient and the absorbed and resecreted nutrient may mix in the gut, preventing quantitative calculation of either. We have developed a surgical model that prevents this complication. Pigs (20-40 kg) were fitted with cannulas in the bile duct, lumen of the duodenum, portal vein, ileocolic vein and jugular vein. After recovery for 6-8 d, pigs were given an oral dose of 9.25 mBq of 54Mn. The flow rate of blood past the portal vein was determined by infusion of P-amino hippuric acid into the ileocolic vein. Absorption was quantified by multiplying the concentration of 54Mn in the portal blood by the flow rate. Biliary excretion was determined by quantitative collection of bile, and previously collected bile was reinfused into the gut lumen. Urine and feces were also quantitatively collected. A postoperative time of 6-8 d was sufficient for pigs to recover from the effects of surgery and anesthesia, as assessed by several measures of metabolic function and food and water intake. True absorption was calculated to be 0.5%. 54Mn in the urine and bile began to increase after 4 d. When the pigs were killed after 12 d, only 0.5% of the 54Mn remained in the carcass. Results of this study show that pigs surgically modified by the described procedure can recover fully and can serve as a model to study intestinal absorption and biliary excretion of nutrients. Furthermore, initial studies using 54Mn showed that the model is applicable to studying Mn metabolism and suggest the need for a more detailed study of Mn absorption and biliary excretion. PMID- 9405584 TI - Endogenous synthesis of arginine plays an important role in maintaining arginine homeostasis in postweaning growing pigs. AB - This study was conducted to determine whether endogenous synthesis of arginine plays a role in regulating arginine homeostasis in postweaning pigs. Pigs were fed a sorghum-based diet containing 0. 98% arginine and were used for studies at 75 d of age (28.4 kg body weight). Mitochondria were prepared from the jejunum and other major tissues for measuring the activities of Delta1-pyrroline-5 carboxylate (P5C) synthase and proline oxidase (enzymes catalyzing P5C synthesis from glutamate and proline, respectively) and of ornithine aminotransferase (OAT) (the enzyme catalyzing the interconversion of P5C into ornithine). For metabolic studies, jejunal enterocytes were incubated at 37 degrees C for 30 min in Krebs Henseleit bicarbonate buffer containing 2 mmol/L L-glutamine, 2 mmol/L L-[U 14C]proline, and 0-200 micromol/L gabaculine (an inhibitor of OAT). The activities of P5C synthase, proline oxidase and OAT were greatest in enterocytes among all of the tissues studied. Incubation of enterocytes with gabaculine resulted in decreases (P < 0.05) in the synthesis of ornithine and citrulline from glutamine and proline. When gabaculine was orally administered to pigs (0.83 mg/kg body weight) to inhibit intestinal synthesis of citrulline from glutamine and proline, plasma concentrations of citrulline (-26%) and arginine (-22%) decreased (P < 0.05), whereas those of alanine (+21%), ornithine (+17%), proline (+107%), taurine (+56%) and branched-chain amino acids (+21-40%) increased (P < 0.05). On the basis of dietary arginine intake and estimated arginine utilization, the endogenous synthesis of arginine in the 28-kg pig provided >/=50.2% of total daily arginine requirement. Taken together, our results suggest an important role for endogenous synthesis of arginine in regulating arginine homeostasis in postweaning growing pigs, as previously shown in neonatal pigs. PMID- 9405585 TI - Intestinal transit and absorption of soy protein in dogs depend on load and degree of protein hydrolysis. AB - Soy protein, in both intact and hydrolyzed forms, is widely used as the nitrogen source in infant and adult formulas. This protein is also consumed in vast quantities worldwide as soybean-based food products. Digestion is the rate limiting step in the assimilation of proteins from the gut. As a result, intestinal transit must be slowed when a higher load of protein is available or when this nutrient is delivered in the intact rather than hydrolyzed form. However, little information is available on the effect of load and degree of hydrolysis of soy protein on intestinal transit and protein absorption. To test the hypothesis that inhibition of intestinal transit and protein absorption depend on the load of soy protein and the state of hydrolysis of this nutrient, we compared intestinal transit and protein absorption in dogs equipped with duodenal and midintestinal fistulas during intestinal perfusion with 0, 50, 100, or 200 g/L solutions of intact soy protein versus 0, 100, 200, 300, or 400 g/L solutions of hydrolyzed soy protein. We found that intestinal transit was slowed in a load-dependent fashion by intact (P < 0.001) and hydrolyzed (P < 0.05) soy protein. Soy protein inhibited intestinal transit more potently in the intact than hydrolyzed form (P < 0.05). A greater amount of protein was absorbed by the proximal half of the small intestine when soy protein was delivered in the hydrolyzed than intact form (P < 0.05), and the efficiency of protein absorption was maintained at a high and nearly constant level of 82.6 to 87.4% for intact soy protein and 89.0 to 92.3% for hydrolyzed soy protein. We conclude that in dogs intestinal transit and absorption of soy protein depend on the load and the degree of protein hydrolysis. PMID- 9405586 TI - Chromic oxide inclusion in the diet does not affect glucose utilization or chromium retention by channel catfish, Ictalurus punctatus. AB - This study was conducted to determine if the level of dietary chromic oxide will affect glucose utilization and tissue chromium retention by channel catfish. Purified diets containing graded levels of supplemental chromic oxide (0, 50, 100, 200, 400, 1000, 5000 and 10,000 mg/kg diet) and glucose as the carbohydrate source were fed to channel catfish fingerlings for 10 wk. Another diet containing dextrin as the carbohydrate source and without chromic oxide supplementation was also fed and served as the control diet. Fish fed the dextrin diet had significantly (P < 0.05) greater weight gain, feed efficiency ratio and protein efficiency ratio but lower plasma glucose concentrations than fish fed the glucose diets irrespective of the level of chromic oxide supplementation. The growth performance and postprandial plasma glucose concentrations of channel catfish fed glucose diets supplemented with various chromic oxide levels were not significantly different. No obvious trends were observed in the whole-body composition of fish fed glucose diets containing various chromic oxide levels. Carbohydrate source or the level of dietary chromic oxide did not significantly affect chromium concentrations in the whole-fish carcass. These results suggest that the level of dietary chromic oxide had no significant effect on glucose utilization or chromium retention by channel catfish. It is suggested that chromic oxide is sufficiently inert to be used as an external marker in digestibility studies in channel catfish. PMID- 9405587 TI - Folate status response to controlled folate intake in pregnant women. AB - A metabolic study (84-d) was conducted to investigate the folate status response of pregnant subjects (n = 12) during their second trimester and nonpregnant controls (n = 12) to folate intakes approximating the current (400 microg/d) and former (800 microg/d) recommended dietary allowance (RDA). The overall goal of the study was to provide metabolic data to assist in the interpretation of the current RDA for folate. Subjects were fed a controlled diet containing 120 +/- 15 microg/d (mean +/- SD) folate and either 330 or 730 microg/d synthetic folic acid. Outcome variables between and within supplementation groups were compared at steady state. Serum folate was higher (P 0.05) were detected in serum folate between pregnant and nonpregnant women within the same supplementation group. Urinary 5-methyl-tetrahydrofolate excretion was greater (P 0.05) in 5-methyl-tetrahydrofolate excretion were detected between pregnant and nonpregnant women within supplementation groups. Differences (P 3Galbeta1 --> 4GlcNAc --> R structure and poly-N acetyllactosamine extension. PMID- 9405607 TI - An essential component of a C-terminal domain phosphatase that interacts with transcription factor IIF in Saccharomyces cerevisiae. AB - One of the essential components of a phosphatase that specifically dephosphorylates the Saccharomyces cerevisiae RNA polymerase II (RPII) large subunit C-terminal domain (CTD) is a novel polypeptide encoded by an essential gene termed FCP1. The Fcp1 protein is localized to the nucleus, and it binds the largest subunit of the yeast general transcription factor IIF (Tfg1). In vitro, transcription factor IIF stimulates phosphatase activity in the presence of Fcp1 and a second complementing fraction. Two distinct regions of Fcp1 are capable of binding to Tfg1, but the C-terminal Tfg1 binding domain is dispensable for activity in vivo and in vitro. Sequence comparison reveals that residues 173-357 of Fcp1 correspond to an amino acid motif present in proteins of unknown function predicted in many organisms. PMID- 9405608 TI - Influenza hemagglutinin is spring-loaded by a metastable native conformation. AB - Enveloped viruses enter cells by protein-mediated membrane fusion. For influenza virus, membrane fusion is regulated by the conformational state of the hemagglutinin (HA) protein, which switches from a native (nonfusogenic) structure to a fusion-active (fusogenic) conformation when exposed to the acidic environment of the cellular endosome. Here we demonstrate that destabilization of HA at neutral pH, with either heat or the denaturant urea, triggers a conformational change that is biochemically indistinguishable from the change triggered by low pH. In each case, the conformational change is coincident with induction of membrane-fusion activity, providing strong evidence that the fusogenic structure is formed. These results indicate that the native structure of HA is trapped in a metastable state and that the fusogenic conformation is released by destabilization of native structure. This strategy may be shared by other enveloped viruses, including those that enter the cell at neutral pH, and could have implications for understanding the membrane-fusion step of HIV infection. PMID- 9405609 TI - Hydrophobic sequence minimization of the alpha-lactalbumin molten globule. AB - The molten globule, a widespread protein-folding intermediate, can attain a native-like backbone topology, even in the apparent absence of rigid side-chain packing. Nonetheless, mutagenesis studies suggest that molten globules are stabilized by some degree of side-chain packing among specific hydrophobic residues. Here we investigate the importance of hydrophobic side-chain diversity in determining the overall fold of the alpha-lactalbumin molten globule. We have replaced all of the hydrophobic amino acids in the sequence of the helical domain with a representative amino acid, leucine. Remarkably, the minimized molecule forms a molten globule that retains many structural features characteristic of a native alpha-lactalbumin fold. Thus, nonspecific hydrophobic interactions may be sufficient to determine the global fold of a protein. PMID- 9405610 TI - Identification and characterization of a human protein kinase related to budding yeast Cdc7p. AB - The Cdc7p protein kinase is essential for the G1/S transition and initiation of DNA replication during the cell division cycle in Saccharomyces cerevisiae. Cdc7p appears to be an evolutionarily conserved protein, since a homolog Hsk1 has been isolated from Schizosaccharomyces pombe. Here, we report the isolation of a human cDNA, HsCdc7, whose product is closely related in sequence to Cdc7p and Hsk1. The HsCdc7 cDNA encodes a protein of 574 amino acids with predicted size of 64 kDa. HsCdc7 contains the conserved subdomains common to all protein-serine/threonine kinases and three "kinase inserts" that are characteristic of Cdc7p and Hsk1. Immune complexes of HsCdc7 from cell lysates were able to phosphorylate histone H1 in vitro. Indirect immunofluorescence staining demonstrated that HsCdc7 protein was predominantly localized in the nucleus. Although the expression levels of HsCdc7 appeared to be constant throughout the cell cycle, the protein kinase activity of HsCdc7 increased during S phase of the cell cycle at approximately the same time as that of Cdk2. These results, together with the functions of Cdc7p in yeast, suggest that HsCdc7 may phosphorylate critical substrate(s) that regulate the G1/S phase transition and/or DNA replication in mammalian cells. PMID- 9405611 TI - The zntA gene of Escherichia coli encodes a Zn(II)-translocating P-type ATPase. AB - The first Zn(II)-translocating P-type ATPase has been identified as the product of o732, a potential gene identified in the sequencing of the Escherichia coli genome. This gene, termed zntA, was disrupted by insertion of a kanamycin gene through homologous recombination. The mutant strain exhibited hypersensitivity to zinc and cadmium salts but not salts of other metals, suggesting a role in zinc homeostasis in E. coli. Everted membrane vesicles from a wild-type strain accumulated 65Zn(II) and 109Cd(II) by using ATP as an energy source. Transport was sensitive to vanadate, an inhibitor of P-type ATPases. Membrane vesicles from the zntA::kan strain did not accumulate those metal ions. Both the sensitive phenotype and transport defect of the mutant were complemented by expression of zntA on a plasmid. PMID- 9405613 TI - Thermodynamic stability of wild-type and mutant p53 core domain. AB - Some 50% of human cancers are associated with mutations in the core domain of the tumor suppressor p53. Many mutations are thought just to destabilize the protein. To assess this and the possibility of rescue, we have set up a system to analyze the stability of the core domain and its mutants. The use of differential scanning calorimetry or spectroscopy to measure its melting temperature leads to irreversible denaturation and aggregation and so is useful as only a qualitative guide to stability. There are excellent two-state denaturation curves on the addition of urea that may be analyzed quantitatively. One Zn2+ ion remains tightly bound in the holo-form of p53 throughout the denaturation curve. The stability of wild type is 6.0 kcal (1 kcal = 4.18 kJ)/mol at 25 degrees C and 9.8 kcal/mol at 10 degrees C. The oncogenic mutants R175H, C242S, R248Q, R249S, and R273H are destabilized by 3.0, 2.9, 1.9, 1.9, and 0.4 kcal/mol, respectively. Under certain denaturing conditions, the wild-type domain forms an aggregate that is relatively highly fluorescent at 340 nm on excitation at 280 nm. The destabilized mutants give this fluorescence under milder denaturation conditions. PMID- 9405612 TI - Kinetic characterization of brush border myosin-I ATPase. AB - Brush border myosin-I (BBM-I) is a single-headed unconventional myosin found in the microvilli of intestinal epithelial cells. We used stopped-flow kinetic analysis to measure the rate and equilibrium constants for several steps in the BBM-I ATPase cycle. We determined the rates for ATP binding to BBM-I and brush border actomyosin-I (actoBBM-I), the rate of actoBBM-I dissociation by ATP, and the rates for the steps in ADP dissociation from actoBBM-I. The rate and equilibrium constants for several of the steps in the actoBBM-I ATPase are significantly different from those of other members of the myosin superfamily. Most notably, dissociation of the actoBBM-I complex by ATP and release of ADP from actoBBM-I are both very slow. The slow rates of these steps may play a role in lengthening the time spent in force-generating states and in limiting the maximal rate of BBM-I motility. In addition, release of ADP from the actoBBM-I complex occurs in at least two steps. This study provides evidence for a member of the myosin superfamily with markedly divergent kinetic behavior. PMID- 9405614 TI - Explanation by the double-metal-ion mechanism of catalysis for the differential metal ion effects on the cleavage rates of 5'-oxy and 5'-thio substrates by a hammerhead ribozyme. AB - In a previous examination using natural all-RNA substrates that contained either a 5'-oxy or 5'-thio leaving group at the cleavage site, we demonstrated that (i) the attack by the 2'-oxygen at C17 on the phosphorus atom is the rate-limiting step only for the substrate that contains a 5'-thio group (R11S) and (ii) the departure of the 5' leaving group is the rate-limiting step for the natural all RNA substrate (R11O) in both nonenzymatic and hammerhead ribozyme-catalyzed reactions; the energy diagrams for these reactions were provided in our previous publication. In this report we found that the rate of cleavage of R11O by a hammerhead ribozyme was enhanced 14-fold when Mg2+ ions were replaced by Mn2+ ions, whereas the rate of cleavage of R11S was enhanced only 2.2-fold when Mg2+ ions were replaced by Mn2+ ions. This result appears to be exactly the opposite of that predicted from the direct coordination of the metal ion with the leaving 5'-oxygen, because a switch in metal ion specificity was not observed with the 5' thio substrate. However, our quantitative analyses based on the previously provided energy diagram indicate that this result is in accord with the double metal-ion mechanism of catalysis. PMID- 9405615 TI - Regulation of dopaminergic pathways by retinoids: activation of the D2 receptor promoter by members of the retinoic acid receptor-retinoid X receptor family. AB - Dopamine is a neuromodulator involved in the control of key physiological functions. Dopamine-dependent signal transduction is activated through the interaction with membrane receptors of the seven-transmembrane domain G protein coupled family. Among them, dopamine D2 receptor is highly expressed in the striatum and the pituitary gland as well as by mesencephalic dopaminergic neurons. Lack of D2 receptors in mice leads to a locomotor parkinsonian-like phenotype and to pituitary tumors. The D2 receptor promoter has characteristics of a housekeeping gene. However, the restricted expression of this gene to particular neurons and cells points to a strict regulation of its expression by cell-specific transcription factors. We demonstrate here that the D2 receptor promoter contains a functional retinoic acid response element. Furthermore, analysis of retinoic acid receptor-null mice supports our finding and shows that in these animals D2 receptor expression is reduced. This finding assigns to retinoids an important role in the control of gene expression in the central nervous system. PMID- 9405616 TI - In vitro selection for altered divalent metal specificity in the RNase P RNA. AB - The ribozyme RNase P absolutely requires divalent metal ions for catalytic function. Multiple Mg2+ ions contribute to the optimal catalytic efficiency of RNase P, and it is likely that the tertiary structure of the ribozyme forms a specific metal-binding pocket for these ions within the active-site. To identify base moieties that contribute to catalytic metal-binding sites, we have used in vitro selection to isolate variants of the Escherichia coli RNase P RNA with altered specificities for divalent metal. RNase P RNA variants with increased activity in Ca2+ were enriched over 18 generations of selection for catalysis in the presence of Ca2+, which is normally disfavored relative to Mg2+. Although a wide spectrum of mutations was found in the generation-18 clones, only a single point mutation was common to all clones: a cytosine-to-uracil transition at position 70 (E. coli numbering) of RNase P. Analysis of the C70U point mutant in a wild-type background confirmed that the identity of the base at position 70 is the sole determinant of Ca2+ selectivity. It is noteworthy that C70 lies within the phylogenetically well conserved J3/4-P4-J2/4 region, previously implicated in Mg2+ binding. Our finding that a single base change is sufficient to alter the metal preference of RNase P is further evidence that the J3/4-P4-J2/4 domain forms a portion of the ribozyme's active site. PMID- 9405617 TI - The anaphase inhibitor of Saccharomyces cerevisiae Pds1p is a target of the DNA damage checkpoint pathway. AB - Inhibition of DNA replication and physical DNA damage induce checkpoint responses that arrest cell cycle progression at two different stages. In Saccharomyces cerevisiae, the execution of both checkpoint responses requires the Mec1 and Rad53 proteins. This observation led to the suggestion that these checkpoint responses are mediated through a common signal transduction pathway. However, because the checkpoint-induced arrests occur at different cell cycle stages, the downstream effectors mediating these arrests are likely to be distinct. We have previously shown that the S. cerevisiae protein Pds1p is an anaphase inhibitor and is essential for cell cycle arrest in mitosis in the presence DNA damage. Herein we show that DNA damage, but not inhibition of DNA replication, induces the phosphorylation of Pds1p. Analyses of Pds1p phosphorylation in different checkpoint mutants reveal that in the presence of DNA damage, Pds1p is phosphorylated in a Mec1p- and Rad9p-dependent but Rad53p-independent manner. Our data place Pds1p and Rad53p on parallel branches of the DNA damage checkpoint pathway. We suggest that Pds1p is a downstream target of the DNA damage checkpoint pathway and that it is involved in implementing the DNA damage checkpoint arrest specifically in mitosis. PMID- 9405618 TI - Wild-type Escherichia coli grows on the chitin disaccharide, N,N' diacetylchitobiose, by expressing the cel operon. AB - We report here that wild-type Escherichia coli can grow on the chitin disaccharide, N,N'-diacetylchitobiose (GlcNAc)2, as the sole source of carbon. Transposon mutants were isolated that were unable to ferment (GlcNAc)2 but grew normally on the monosaccharide GlcNAc. One such mutant was used to screen a wild type E. coli genomic cosmid library for restoration of (GlcNAc)2 fermentation. A partial sequence analysis of the isolated fragment mapped the clone to the (previously sequenced) E. coli genome between 39.0 and 39.2 min. The nucleotide ORFs at this region had been previously assigned to code for a "cryptic" cellobiose utilization (cel) operon. We report here, however, that functional analysis of the operon, including growth and chemotaxis, reveal that it encodes a set of proteins that are not cryptic, but are induced by (GlcNAc)2 and catabolize the disaccharide. We therefore propose to rename the cel operon as the chb (N,N' diacetylchitobiose) operon, with the letter designation of the genes of the operon to be reassigned consistent with the nomenclature based on functional characterization of the gene products as follows: celA to chbB, celB to chbC, celC to chbA, celD to chbR, and celF to chbF. Furthermore, sequencing evidence indicates that the operon contains an additional gene of unknown function to be designated as chbG. Thus, the overall gene sequence is to be named chbBCARFG. PMID- 9405619 TI - Cloning and functional characterization of mouse IkappaBepsilon. AB - The biological activity of the transcription factor NF-kappaB is mainly controlled by the IkappaB proteins IkappaBalpha and IkappaBbeta, which restrict NF-kappaB in the cytoplasm and enter the nucleus where they terminate NF-kappaB dependent transcription. In this paper we describe the cloning and functional characterization of mouse IkappaBepsilon. Mouse IkappaBepsilon contains 6 ankyrin repeats required for its interaction with the Rel proteins and is expressed in different cell types where we found that it is up-regulated by NF-kappaB inducers, as is the case for IkappaBalpha and human IkappaBepsilon. IkappaBepsilon functions as a bona fide IkappaB protein by restricting Rel proteins in the cytoplasm and inhibiting their in vitro DNA binding activity. Surprisingly, IkappaBepsilon did not inhibit transcription of genes regulated by the p50/p65 heterodimer efficiently, such as the human interferon-beta gene. However, IkappaBepsilon was a strong inhibitor of interleukin-8 expression, a gene known to be regulated by p65 homodimers. In addition, IkappaBepsilon appears to function predominantly in the cytoplasm to sequester p65 homodimers, in contrast with the other two members of the family, IkappaBalpha and IkappaBbeta, which also function in the nucleus to terminate NF-kappaB-dependent transcriptional activation. PMID- 9405620 TI - The replication initiation protein of the broad-host-range plasmid RK2 is activated by the ClpX chaperone. AB - Initiation and control of replication of the broad-host-range plasmid RK2 requires two plasmid-encoded elements, the replication origin (oriV) and the initiation protein TrfA. Purified TrfA is largely in the form of a dimer; however, only the monomeric form of the protein can bind specifically to the direct repeats (iterons) at the RK2 origin. The largely dimeric form of wild-type TrfA is inactive in the initiation of replication of RK2 in an in vitro replication system reconstituted from purified components. However, preincubation of the TrfA protein with the ClpX molecular chaperone isolated from Escherichia coli activates the initiator protein for replication in the purified system. We further observed that ClpX, in an ATP-dependent reaction, greatly increases the proportion of TrfA monomers and, therefore, the ability of this protein to bind to iterons localized within RK2 origin. Finally, a copy-up mutant of the TrfA protein which is largely in the monomer form is active in the reconstituted in vitro replication system, and its activity is not affected by ClpX. PMID- 9405621 TI - Archaeal-type lysyl-tRNA synthetase in the Lyme disease spirochete Borrelia burgdorferi. AB - Lysyl-tRNAs are essential for protein biosynthesis by ribosomal mRNA translation in all organisms. They are synthesized by lysyl-tRNA synthetases (EC 6.1.1.6), a group of enzymes composed of two unrelated families. In bacteria and eukarya, all known lysyl-tRNA synthetases are subclass IIc-type aminoacyl-tRNA synthetases, whereas some archaea have been shown to contain an unrelated class I-type lysyl tRNA synthetase. Examination of the preliminary genomic sequence of the bacterial pathogen Borrelia burgdorferi, the causative agent of Lyme disease, indicated the presence of an open reading frame with over 55% similarity at the amino acid level to archaeal class I-type lysyl-tRNA synthetases. In contrast, no coding region with significant similarity to any class II-type lysyl-tRNA synthetase could be detected. Heterologous expression of this open reading frame in Escherichia coli led to the production of a protein with canonical lysyl-tRNA synthetase activity in vitro. Analysis of B. burgdorferi mRNA showed that the lysyl-tRNA synthetase-encoding gene is highly expressed, confirming that B. burgdorferi contains a functional class I-type lysyl-tRNA synthetase. The detection of an archaeal-type lysyl-tRNA synthetase in B. burgdorferi and other pathogenic spirochetes, but not to date elsewhere in bacteria or eukarya, indicates that the gene that encodes this enzyme has a common origin with its orthologue from the archaeal kingdom. This difference between the lysyl-tRNA synthetases of spirochetes and their hosts may be readily exploitable for the development of anti-spirochete therapeutics. PMID- 9405622 TI - RGS2/G0S8 is a selective inhibitor of Gqalpha function. AB - RGS (regulators of G protein signaling) proteins are GTPase activating proteins that inhibit signaling by heterotrimeric G proteins. All RGS proteins studied to date act on members of the Gialpha family, but not Gsalpha or G12alpha. RGS4 regulates Gialpha family members and Gqalpha. RGS2 (G0S8) is exceptional because the G proteins it regulates have not been identified. We report that RGS2 is a selective and potent inhibitor of Gqalpha function. RGS2 selectively binds Gqalpha, but not other Galpha proteins (Gi, Go, Gs, G12/13) in brain membranes; RGS4 binds Gqalpha and Gialpha family members. RGS2 binds purified recombinant Gqalpha, but not Goalpha, whereas RGS4 binds either. RGS2 does not stimulate the GTPase activities of Gsalpha or Gialpha family members, even at a protein concentration 3000-fold higher than is sufficient to observe effects of RGS4 on Gialpha family members. In contrast, RGS2 and RGS4 completely inhibit Gq-directed activation of phospholipase C in cell membranes. When reconstituted with phospholipid vesicles, RGS2 is 10-fold more potent than RGS4 in blocking Gqalpha directed activation of phospholipase Cbeta1. These results identify a clear physiological role for RGS2, and describe the first example of an RGS protein that is a selective inhibitor of Gqalpha function. PMID- 9405623 TI - Inhibition of mRNA export in vertebrate cells by nuclear export signal conjugates. AB - Leucine-rich nuclear export signals (NESs) are recognized by the NES receptor exportin 1 and are central to the export of multiple shuttling proteins and RNAs. The export of messenger RNA in vertebrates was, however, thought to occur by a different pathway, because inhibition by injection of a synthetic Rev NES conjugate could not be demonstrated. Here we find that peptide conjugates composed of the NES of either protein kinase A inhibitor protein (PKI) or the HIV 1 Rev protein, when coupled to human serum albumin, are potent inhibitors of mRNA and small nuclear RNA export. These results provide direct evidence that mRNA export in vertebrates depends on interactions between an NES and its cognate NES receptors. PKI NES conjugates are significantly more efficient at inhibiting RNA export than are REV NES conjugates, indicating that different NESs may have different abilities to promote protein and RNA export. Surprisingly, an expected control conjugate containing the mutant Rev NES sequence M10 strongly inhibited the export of intronless dihydrofolate reductase mRNA. Nuclear injection of NES peptide conjugates led to mislocalization to the nucleus of 10-20% of the cytoplasmic Ran GTPase-binding protein (RanBP1) indicating that RanBP1 shuttles between the nucleus and the cytoplasm via an NES pathway. These results demonstrate that in vertebrates the export of mRNA, like that of small nuclear RNA, 5S rRNA, and transport factors such as RanBP1, employs NES-mediated molecular machinery. PMID- 9405624 TI - Cloning and characterization of a corepressor and potential component of the nuclear hormone receptor repression complex. AB - Nuclear hormone receptors are potent repressors of transcription in the unliganded state. We describe here the cloning of a nuclear receptor corepressor that we call SUN-CoR (Small Unique Nuclear receptor CoRepressor), which shows no homology to previously described nuclear hormone receptor corepressors, N-CoR, or SMRT. SUN-CoR is a highly basic, 16-kDa nuclear protein that is expressed at high levels in adult tissues and is induced during adipocyte and myogenic differentiation. SUN-CoR potentiates transcriptional repression by thyroid hormone receptor and RevErb in vivo, represses transcription when fused to a heterologous DNA binding domain, and interacts with RevErb as well as with thyroid hormone receptor in vitro. SUN-CoR also interacts with N-CoR and SMRT in vitro and with endogenous N-CoR in cells. We conclude that SUN-CoR is a corepressor and may function as an additional component of the complex involved in transcriptional repression by unliganded and orphan nuclear hormone receptors. PMID- 9405625 TI - Chemical complementation identifies a proton acceptor for redox-active tyrosine D in photosystem II. AB - Through the use of site-directed mutagenesis and chemical rescue, we have identified the proton acceptor for redox-active tyrosine D in photosystem II (PSII). Effects of chemical rescue on the tyrosyl radical were monitored by EPR spectroscopy. We also have acquired the Fourier-transform infrared (FT-IR) spectrum associated with the oxidation of tyrosine D and concomitant protonation of the acceptor. Mutant and isotopically labeled PSII samples are used to assign vibrational lines in the 3,600-3,100 cm-1 region to N-H modes of His-189 in the D2 polypeptide. When His-189 in D2 is changed to a leucine (HL189D2) in PSII, dramatic alterations of both EPR and FT-IR spectra are observed. When imidazole is introduced into HL189D2 samples, results from both EPR and FT-IR spectroscopy argue that imidazole is functionally reconstituted into an accessible pocket and that imidazole acts as a chemical mimic for His-189. Small perturbations of EPR and FT-IR spectra are consistent with access to this pocket in wild-type PSII, as well. Structures of the analogous site in bacterial reaction centers suggest that an accessible pocket, large enough to contain imidazole, is bordered by tyrosine D and His-189 in the D2 polypeptide. These data provide evidence that His-189 in the D2 polypeptide of PSII acts as a proton acceptor for redox-active tyrosine D and that proton transfer to the imidazole ring facilitates the efficient oxidation/reduction of tyrosine D. PMID- 9405626 TI - Virus-sized self-assembling lamellar complexes between plasmid DNA and cationic micelles promote gene transfer. AB - Gene therapy is based on the vectorization of genes to target cells and their subsequent expression. Cationic amphiphile-mediated delivery of plasmid DNA is the nonviral gene transfer method most often used. We examined the supramolecular structure of lipopolyamine/plasmid DNA complexes under various condensing conditions. Plasmid DNA complexation with lipopolyamine micelles whose mean diameter was 5 nm revealed three domains, depending on the lipopolyamine/plasmid DNA ratio. These domains respectively corresponded to negatively, neutrally, and positively charged complexes. Transmission electron microscopy and x-ray scattering experiments on complexes originating from these three domains showed that although their morphology depends on the lipopolyamine/plasmid DNA ratio, their particle structure consists of ordered domains characterized by even spacing of 80 A, irrespective of the lipid/DNA ratio. The most active lipopolyamine/DNA complexes for gene transfer were positively charged. They were characterized by fully condensed DNA inside spherical particles (diameter: 50 nm) sandwiched between lipid bilayers. These results show that supercoiled plasmid DNA is able to transform lipopolyamine micelles into a supramolecular organization characterized by ordered lamellar domains. PMID- 9405627 TI - Torsional directed walks, entropic elasticity, and DNA twist stiffness. AB - DNA and other biopolymers differ from classical polymers because of their torsional stiffness. This property changes the statistical character of their conformations under tension from a classical random walk to a problem we call the "torsional directed walk." Motivated by a recent experiment on single lambda-DNA molecules [Strick, T. R., Allemand, J.-F., Bensimon, D., Bensimon, A. & Croquette, V. (1996) Science 271, 1835-1837], we formulate the torsional directed walk problem and solve it analytically in the appropriate force regime. Our technique affords a direct physical determination of the microscopic twist stiffness C and twist-stretch coupling D relevant for DNA functionality. The theory quantitatively fits existing experimental data for relative extension as a function of overtwist over a wide range of applied force; fitting to the experimental data yields the numerical values C = 120 nm and D = 50 nm. Future experiments will refine these values. We also predict that the phenomenon of reduction of effective twist stiffness by bend fluctuations should be testable in future single-molecule experiments, and we give its analytic form. PMID- 9405628 TI - Flickering fusion pores comparable with initial exocytotic pores occur in protein free phospholipid bilayers. AB - For the act of membrane fusion, there are two competing, mutually exclusive molecular models that differ in the structure of the initial pore, the pathway for ionic continuity between formerly separated volumes. Because biological "fusion pores" can be as small as ionic channels or gap junctions, one model posits a proteinaceous initial fusion pore. Because biological fusion pore conductance varies widely, another model proposes a lipidic initial pore. We have found pore opening and flickering during the fusion of protein-free phospholipid vesicles with planar phospholipid bilayers. Fusion pore formation appears to follow the coalescence of contacting monolayers to create a zone of hemifusion where continuity between the two adherent membranes is lipidic, but not aqueous. Hypotonic stress, causing tension in the vesicle membrane, promotes complete fusion. Pores closed soon after opening (flickering), and the distribution of fusion pore conductance appears similar to the distribution of initial fusion pores in biological fusion. Because small flickering pores can form in the absence of protein, the existence of small pores in biological fusion cannot be an argument in support of models based on proteinaceous pores. Rather, these results support the model of a lipidic fusion pore developing within a hemifused contact site. PMID- 9405629 TI - Folding thermodynamics of a model three-helix-bundle protein. AB - The calculated folding thermodynamics of a simple off-lattice three-helix-bundle protein model under equilibrium conditions shows the experimentally observed protein transitions: a collapse transition, a disordered-to-ordered globule transition, a globule to native-state transition, and the transition from the active native state to a frozen inactive state. The cooperativity and physical origin of the various transitions are explored with a single "optimization" parameter and characterized with the Lindemann criterion for liquid versus solid state dynamics. Below the folding temperature, the model has a simple free energy surface with a single basin near the native state; the surface is similar to that calculated from a simulation of the same three-helix-bundle protein with an all atom representation [Boczko, E. M. & Brooks III, C. L. (1995) Science 269, 393 396]. PMID- 9405630 TI - Absence of a barrier to backwards rotation of the bacterial flagellar motor demonstrated with optical tweezers. AB - A cell of the bacterium Escherichia coli was tethered covalently to a glass coverslip by a single flagellum, and its rotation was stopped by using optical tweezers. The tweezers acted directly on the cell body or indirectly, via a trapped polystyrene bead. The torque generated by the flagellar motor was determined by measuring the displacement of the laser beam on a quadrant photodiode. The coverslip was mounted on a computer-controlled piezo-electric stage that moved the tether point in a circle around the center of the trap so that the speed of rotation of the motor could be varied. The motor generated approximately 4500 pN nm of torque at all angles, regardless of whether it was stalled, allowed to rotate very slowly forwards, or driven very slowly backwards. This argues against models of motor function in which rotation is tightly coupled to proton transit and back-transport of protons is severely limited. PMID- 9405631 TI - Subunit structure of the mammalian exocyst complex. AB - The exocyst is a protein complex required for the late stages of secretion in yeast. Unlike the SNAREs (SNAP receptors), important secretory proteins that are broadly distributed on the target membrane, the exocyst is specifically located at sites of vesicle fusion. We have isolated cDNAs encoding the rexo70, rsec5, and rsec15 subunits of the mammalian complex. The amino acid sequences encoded by these genes are between 21% and 24% identical to their yeast homologs. All three genes are broadly expressed and multiple transcripts are observed for rexo70 and rsec15. Characterization of cDNAs encoding the 84-kDa subunit of the mammalian complex revealed a novel protein. mAbs were generated to the mammalian rsec6 subunit of the exocyst complex. rsec6 immunoreactivity is found in a punctate distribution at terminals of PC12 cell processes at or near sites of granule exocytosis. PMID- 9405632 TI - Generation of secretable and nonsecretable interleukin 15 isoforms through alternate usage of signal peptides. AB - Two isoforms of human interleukin 15 (IL-15) exist. One isoform has a shorter putative signal peptide (21 amino acids) and its transcript shows a tissue distribution pattern that is distinct from that of the alternative IL-15 isoform with a 48-aa signal peptide. The 21-aa signal isoform is preferentially expressed in tissues such as testis and thymus. Experiments using different combinations of signal peptides and mature proteins (IL-2, IL-15, and green fluorescent protein) showed that the short signal peptide regulates the fate of the mature protein by controlling the intracellular trafficking to nonendoplasmic reticulum sites, whereas the long signal peptide both regulates the rate of protein translation and functions as a secretory signal peptide. As a consequence, the IL-15 associated with the short signal peptide is not secreted, but rather is stored intracellularly, appearing in the nucleus and cytoplasmic components. Such production of an intracellular lymphokine is not typical of other soluble interleukin systems, suggesting a biological function for IL-15 as an intracellular molecule. PMID- 9405633 TI - Otogelin: a glycoprotein specific to the acellular membranes of the inner ear. AB - Efforts to identify the specific components of the mammalian inner ear have been hampered by the small number of neuroepithelial cells and the variety of supporting cells. To circumvent these difficulties, we used a PCR-based subtractive method on cDNA from 2-day-old mouse cochlea. A cDNA encoding a predicted 2910-amino acid protein related to mucin has been isolated. Several lines of evidence indicate, however, that this protein does not undergo the O glycosylation characteristic to mucins. As confirmed by immunocytochemistry and biochemical experiments, this protein is specific to the inner ear. Immunohistofluorescence labeling showed that this protein is a component of all the acellular membranes of the inner ear: i.e., the tectorial membrane of the cochlea, the otoconial and accessory membranes of the utricule and saccule, the cupula of the semicircular canals, and a previously undescribed acellular material covering the otoconia of the saccule. The protein has been named otogelin with reference to its localization. A variety of nonsensory cells located underneath these membranes could be identified as synthesizing otogelin. Finally, this study revealed a maturation process of the tectorial membrane, as evidenced by the progressive organization of otogelin labeling into thick and spaced radial fiber-like structures. PMID- 9405634 TI - D-myo-Inositol 1,4,5,6-tetrakisphosphate produced in human intestinal epithelial cells in response to Salmonella invasion inhibits phosphoinositide 3-kinase signaling pathways. AB - Several inositol-containing compounds play key roles in receptor-mediated cell signaling events. Here, we describe a function for a specific inositol polyphosphate, D-myo-inositol 1,4,5,6-tetrakisphosphate [Ins(1,4,5,6)P4], that is produced acutely in response to a receptor-independent process. Thus, infection of intestinal epithelial cells with the enteric pathogen Salmonella, but not with other invasive bacteria, induced a multifold increase in Ins(1,4,5,6)P4 levels. To define a specific function of Ins(1,4,5,6)P4, a membrane-permeant, hydrolyzable ester was used to deliver it to the intracellular compartment, where it antagonized epidermal growth factor (EGF)-induced inhibition of calcium mediated chloride (Cl-) secretion (CaMCS) in intestinal epithelia. This EGF function is likely mediated through a phosphoinositide 3-kinase (PtdIns3K) dependent mechanism because the EGF effects are abolished by wortmannin, and three different membrane-permeant esters of the PtdIns3K product phosphatidylinositol 3,4,5-trisphosphate mimicked the EGF effect on CaMCS. We further demonstrate that Ins(1,4,5,6)P4 antagonized EGF signaling downstream of PtdIns3K because Ins(1,4,5, 6)P4 interfered with the PtdInsP3 effect on CaMCS without affecting PtdIns3K activity. Thus, elevation of Ins(1,4,5,6)P4 in Salmonella-infected epithelia may promote Cl- flux by antagonizing EGF inhibition mediated through PtdIns3K and PtdInsP3. PMID- 9405635 TI - Protein kinase A activity is required for the budding of constitutive transport vesicles from the trans-Golgi network. AB - We have examined the role played by protein kinase A (PKA) in vesicle-mediated protein transport from the trans-Golgi network (TGN) to the cell surface. In vivo this transport step was inhibited by inhibitors of PKA catalytic subunits (C-PKA) such as the compound known as H89 and a myristoylated form of the inhibitory peptide sequence contained in the thermostable PKA inhibitor. Inhibition by H89 occurred at an early stage during the transfer of vesicular stomatitis virus G glycoprotein from the TGN to the cell surface. Reversal from this inhibition correlated with a transient increase in the number of free coated vesicles in the Golgi area. Vesicle budding from the TGN was studied in vitro using vesicular stomatitis virus-infected, permeabilized cells. Addition to this assay of C-PKA stimulated vesicle release while it was suppressed by PKA inhibitory peptide, H89, and antibody against C-PKA. Furthermore, vesicle release was decreased when PKA-depleted cytosol was used and restored by addition of C-PKA. These results indicate a regulatory role for PKA activity in the production of constitutive transport vesicles from the TGN. PMID- 9405636 TI - The mechanism of Golgi segregation during mitosis is cell type-specific. AB - Golgi membranes in Drosophila embryos and tissue culture cells are found as discrete units dispersed in the cytoplasm. We provide evidence that Golgi membranes do not undergo any dramatic change in their organization during the rapid mitotic divisions of the nuclei in the syncitial embryo or during cell division postcellularization. By contrast, in Drosophila tissue culture cells, the Golgi membranes undergo complete fragmentation during mitosis. Our studies show that the mechanism of Golgi partitioning during cell division is cell type specific. PMID- 9405637 TI - Proper sorting of the cation-dependent mannose 6-phosphate receptor in endosomes depends on a pair of aromatic amino acids in its cytoplasmic tail. AB - The 67-amino acid cytoplasmic tail of the cation-dependent mannose 6-phosphate receptor (CD-MPR) contains a signal(s) that prevents the receptor from entering lysosomes where it would be degraded. To identify the key residues required for proper endosomal sorting, we analyzed the intracellular distribution of mutant forms of the receptor by Percoll density gradients. A receptor with a Trp19 --> Ala substitution in the cytoplasmic tail was highly missorted to lysosomes whereas receptors with either Phe18 --> Ala or Phe13 --> Ala mutations were partially defective in avoiding transport to lysosomes. Analysis of double and triple mutants confirmed the key role of Trp19 for sorting of the CD-MPR in endosomes, with Phe18, Phe13, and several neighboring residues contributing to this function. The addition of the Phe18-Trp19 motif of the CD-MPR to the cytoplasmic tail of the lysosomal membrane protein Lamp1 was sufficient to partially impair its delivery to lysosomes. Replacing Phe18 and Trp19 with other aromatic amino acids did not impair endosomal sorting of the CD-MPR, indicating that two aromatic residues located at these positions are sufficient to prevent the receptor from trafficking to lysosomes. However, alterations in the spacing of the diaromatic amino acid sequence relative to the transmembrane domain resulted in receptor accumulation in lysosomes. These findings indicate that the endosomal sorting of the CD-MPR depends on the correct presentation of a diaromatic amino acid-containing motif in its cytoplasmic tail. Because a diaromatic amino acid sequence is also present in the cytoplasmic tail of other receptors known to be internalized from the plasma membrane, this feature may prove to be a general determinant for endosomal sorting. PMID- 9405638 TI - A role for the epithelial-cell-specific tyrosine kinase Sik during keratinocyte differentiation. AB - Sik, the mouse homologue of the breast tumor kinase Brk, is expressed in differentiating cells of the gastrointestinal tract and skin. We examined expression and activity of Sik in primary mouse keratinocytes and a mouse embryonic keratinocyte cell line (EMK). Calcium-induced differentiation of these cells has been shown to be accompanied by the activation of tyrosine kinases and rapid phosphorylation of a 65-kDa GTPase-activating protein (GAP)-associated protein (GAP-A.p65). We demonstrate that Sik is activated within 2 min after calcium addition in primary keratinocytes and EMK cells. In EMK cells, Sik binds GAP-A.p65, and this interaction is mediated by the Sik Src homology 2 domain. Although Sik directly complexes with GAP-A.p65, overexpression of wild-type or kinase defective Sik in EMK cells does not lead to detectable changes in GAP A.p65 phosphorylation. These data suggest that Sik is not responsible for phosphorylation of GAP-A.p65. GAP-A. p65 may act as an adapter protein, bringing Sik into proximity of an unidentified substrate. Overexpression of Sik in EMK cells results in increased expression of filaggrin during differentiation, supporting a role for Sik in differentiation. PMID- 9405639 TI - Localization of a constitutively active, phagocyte-like NADPH oxidase in rabbit aortic adventitia: enhancement by angiotensin II. AB - Superoxide anion (O2-) plays a key role in the endogenous suppression of endothelium-derived nitric oxide (NO) bioactivity and has been implicated in the development of hypertension. In previous studies, we found that O2- is produced predominantly in the adventitia of isolated rabbit aorta and acts as a barrier to NO. In the present studies, we characterize the enzyme responsible for O2- production in the adventitia and show that this enzyme is a constitutively active NADPH oxidase with similar composition as the phagocyte NADPH oxidase. Constitutive O2--generating activity was localized to aortic adventitial fibroblasts and was enhanced by the potent vasoconstrictor angiotensin II. Immunohistochemistry of aortic sections demonstrated the presence of p22(phox), gp91(phox), p47(phox), and p67(phox) localized exclusively in rabbit aortic adventitia, coincident with the site of staining for O2- production. Furthermore, immunodepletion of p67(phox) from adventitial fibroblast particulates resulted in the loss of NADPH oxidase activity, which could be restored by the addition of recombinant p67(phox). Further study into the regulation of this adventitial source of O2- is important in elucidating the mechanisms regulating the bioactivity of NO and may contribute to our understanding of the pathogenesis of hypertension. PMID- 9405640 TI - Receptors induce chemotaxis by releasing the betagamma subunit of Gi, not by activating Gq or Gs. AB - Many chemoattractants cause chemotaxis of leukocytes by stimulating a structurally distinct class of G protein-coupled receptors. To identify receptor functions required for chemotaxis, we studied chemotaxis in HEK293 cells transfected with receptors for nonchemokine ligands or for interleukin 8 (IL-8), a classical chemokine. In gradients of the appropriate agonist, three nonchemokine Gi-coupled receptors (the D2 dopamine receptor and opioid mu and delta receptors) mediated chemotaxis; the beta2-adrenoreceptor and the M3 muscarinic receptor, which couple respectively to Gs and Gq, did not mediate chemotaxis. A mutation deleting 31 C-terminal amino acids from the IL-8 receptor type B quantitatively impaired chemotaxis and agonist-induced receptor internalization, but not inhibition of adenylyl cyclase or stimulation of mitogen activated protein kinase. To probe the possible relation between receptor internalization and chemotaxis, we used two agonists of the mu-opioid receptor. Morphine and etorphine elicited quantitatively similar chemotaxis, but only etorphine induced receptor internalization. Overexpression of two betagamma sequestering proteins (betaARK-ct and alphat) prevented IL-8 receptor type B mediated chemotaxis but did not affect inhibition of adenylyl cyclase by IL-8. We conclude that: (i) Nonchemokine Gi-coupled receptors can mediate chemotaxis. (ii) Gi activation is necessary but probably not sufficient for chemotaxis. (iii) Chemotaxis does not require receptor internalization. (iv) Chemotaxis requires the release of free betagamma subunits. PMID- 9405641 TI - Chemotaxis in a lymphocyte cell line transfected with C-C chemokine receptor 2B: evidence that directed migration is mediated by betagamma dimers released by activation of Galphai-coupled receptors. AB - Chemotaxis is mediated by activation of seven-transmembrane domain, G protein coupled receptors, but the signal transduction pathways leading to chemotaxis are poorly understood. To identify G proteins that signal the directed migration of cells, we stably transfected a lymphocyte cell line (300-19) with G protein coupled receptors that couple exclusively to Galphaq (the m3 muscarinic receptor), Galphai (the kappa-opioid receptor), and Galphas (the beta-adrenergic receptor), as well as the human thrombin receptor (PAR-1) and the C-C chemokine receptor 2B. Cells expressing receptors that coupled to Galphai, but not to Galphaq or Galphas, migrated in response to a concentration gradient of the appropriate agonist. Overexpression of Galpha transducin, which binds to and inactivates free Gbetagamma dimers, completely blocked chemotaxis although having little or no effect on intracellular calcium mobilization or other measures of cell signaling. The identification of Gbetagamma dimers as a crucial intermediate in the chemotaxis signaling pathway provides further evidence that chemotaxis of mammalian cells has important similarities to polarized responses in yeast. We conclude that chemotaxis is dependent on activation of Galphai and the release of Gbetagamma dimers, and that Galphai-coupled receptors not traditionally associated with chemotaxis can mediate directed migration when they are expressed in hematopoietic cells. PMID- 9405642 TI - Phosphorylation of the immunosuppressant FK506-binding protein FKBP52 by casein kinase II: regulation of HSP90-binding activity of FKBP52. AB - FKBP52 (HSP56, p59, HBI) is the 59-kDa immunosuppressant FK506-binding protein and has peptidyl prolyl isomerase as well as a chaperone-like activity in vitro. FKBP52 associates with the heat shock protein HSP90 and is included in the steroid hormone receptor complexes in vivo. FKBP52 possesses a well conserved phosphorylation site for casein kinase II (CK2) that was previously shown to be associated with HSP90. Here we examined whether FKBP52 is phosphorylated by CK2 both in vivo and in vitro. Recombinant rabbit FKBP52 was phosphorylated by purified CK2. We expressed and purified deletion mutants of FKBP52 to determine the site(s) phosphorylated by CK2. Thr-143 in the hinge I region was identified as the major phosphorylation site for CK2. A synthetic peptide corresponding to this region was phosphorylated by CK2, and the peptide competitively inhibited the phosphorylation of other substrates by CK2. The [32P]phosphate labeling of FKBP52-expressing cells revealed that the same site is also phosphorylated in vivo. FK506 binding to FKBP52 did not affect the phosphorylation by CK2 and, conversely, the FK506-binding activity of FKBP52 was not affected by the phosphorylation. Most importantly, CK2-phosphorylated FKBP52 did not bind to HSP90. These results indicate that CK2 phosphorylates FKBP52 both in vitro and in vivo and thus may regulate the protein composition of chaperone-containing complexes such as those of steroid receptors and certain protein kinases. PMID- 9405644 TI - Targeted disruption of the murine dihydrolipoamide dehydrogenase gene (Dld) results in perigastrulation lethality. AB - The Dld gene product, known as dihydrolipoamide dehydrogenase or the E3 component, catalyzes the oxidation of dihydrolipoyl moieties of four mitochondrial multienzyme complexes: pyruvate dehydrogenase, alpha-ketoglutarate dehydrogenase, branched-chain alpha-ketoacid dehydrogenase, and the glycine cleavage system. Deficiency of E3 activity in humans results in various degrees of neurological dysfunction and organic acidosis caused by accumulation of branched-chain amino acids and lactic acid. In this study, we have introduced a null mutation into the murine Dld gene (Dldtm1mjp). The heterozygous animals are shown to have approximately half of wild-type activity levels for E3 and all affected multienzyme complexes but are phenotypically normal. In contrast, the Dld-/- class dies prenatally with apparent developmental delay at 7.5 days postcoitum followed by resorption by 9.5 days postcoitum. The Dld-/- embryos cease to develop at a time shortly after implantation into the uterine wall when most of the embryos have begun to gastrulate. This null phenotype provides in vivo evidence for the requirement of a mitochondrial oxidative pathway during the perigastrulation period. Furthermore, the early prenatal lethal condition of the complete deficiency state may explain the low incidence of detectable cases of E3 deficiency in humans. PMID- 9405643 TI - Aneuploidy correlated 100% with chemical transformation of Chinese hamster cells. AB - Aneuploidy or chromosome imbalance is the most massive genetic abnormality of cancer cells. It used to be considered the cause of cancer when it was discovered more than 100 years ago. Since the discovery of the gene, the aneuploidy hypothesis has lost ground to the hypothesis that mutation of cellular genes causes cancer. According to this hypothesis, cancers are diploid and aneuploidy is secondary or nonessential. Here we reexamine the aneuploidy hypothesis in view of the fact that nearly all solid cancers are aneuploid, that many carcinogens are nongenotoxic, and that mutated genes from cancer cells do not transform diploid human or animal cells. By regrouping the gene pool-as in speciation aneuploidy inevitably will alter many genetic programs. This genetic revolution can explain the numerous unique properties of cancer cells, such as invasiveness, dedifferentiation, distinct morphology, and specific surface antigens, much better than gene mutation, which is limited by the conservation of the existing chromosome structure. To determine whether aneuploidy is a cause or a consequence of transformation, we have analyzed the chromosomes of Chinese hamster embryo (CHE) cells transformed in vitro. This system allows (i) detection of transformation within 2 months and thus about 5 months sooner than carcinogenesis and (ii) the generation of many more transformants per cost than carcinogenesis. To minimize mutation of cellular genes, we have used nongenotoxic carcinogens. It was found that 44 out of 44 colonies of CHE cells transformed by benz[a]pyrene, methylcholanthrene, dimethylbenzanthracene, and colcemid, or spontaneously were between 50 and 100% aneuploid. Thus, aneuploidy originated with transformation. Two of two chemically transformed colonies tested were tumorigenic 2 months after inoculation into hamsters. The cells of transformed colonies were heterogeneous in chromosome number, consistent with the hypothesis that aneuploidy can perpetually destabilize the chromosome number because it unbalances the elements of the mitotic apparatus. Considering that all 44 transformed colonies analyzed were aneuploid, and the early association between aneuploidy, transformation, and tumorigenicity, we conclude that aneuploidy is the cause rather than a consequence of transformation. PMID- 9405645 TI - Conserved biological function between Pax-2 and Pax-5 in midbrain and cerebellum development: evidence from targeted mutations. AB - The development of two major subdivisions of the vertebrate nervous system, the midbrain and the cerebellum, is controlled by signals emanating from a constriction in the neural primordium called the midbrain/hindbrain organizer (Joyner, A. L. (1996) Trends Genet. 12, 15-201). The closely related transcription factors Pax-2 and Pax-5 exhibit an overlapping expression pattern very early in the developing midbrain/hindbrain junction. Experiments carried out in fish (Krauss, S., Maden, M., Holder, N. & Wilson, S. W. (1992) Nature (London) 360, 87-89) with neutralizing antibodies against Pax-b, the orthologue of Pax-2 in mouse, placed this gene family in an regulatory cascade necessary for the development of the midbrain and the cerebellum. The targeted mutation of Pax-5 has been reported to have only slight effects in the development of structures derived from the isthmic constriction, whereas the Pax-2 null mutant mice show a background-dependent phenotype with varying penetrance. To test a possible redundant function between Pax-2 and Pax-5 we analyzed the brain phenotypes of mice expressing different dosages of both genes. Our results demonstrate a conserved biological function of both proteins in midbrain/hindbrain regionalization. Additionally, we show that one allele of Pax-2, but not Pax-5, is necessary and sufficient for midbrain and cerebellum development in C57BL/6 mice. PMID- 9405647 TI - Benthic-pelagic links and rocky intertidal communities: bottom-up effects on top down control? AB - Insight into the dependence of benthic communities on biological and physical processes in nearshore pelagic environments, long considered a "black box," has eluded ecologists. In rocky intertidal communities at Oregon coastal sites 80 km apart, differences in abundance of sessile invertebrates, herbivores, carnivores, and macrophytes in the low zone were not readily explained by local scale differences in hydrodynamic or physical conditions (wave forces, surge flow, or air temperature during low tide). Field experiments employing predator and herbivore manipulations and prey transplants suggested top-down (predation, grazing) processes varied positively with bottom-up processes (growth of filter feeders, prey recruitment), but the basis for these differences was unknown. Shore-based sampling revealed that between-site differences were associated with nearshore oceanographic conditions, including phytoplankton concentration and productivity, particulates, and water temperature during upwelling. Further, samples taken at 19 sites along 380 km of coastline suggested that the differences documented between two sites reflect broader scale gradients of phytoplankton concentration. Among several alternative explanations, a coastal hydrodynamics hypothesis, reflecting mesoscale (tens to hundreds of kilometers) variation in the interaction between offshore currents and winds and continental shelf bathymetry, was inferred to be the primary underlying cause. Satellite imagery and offshore chlorophyll-a samples are consistent with the postulated mechanism. Our results suggest that benthic community dynamics can be coupled to pelagic ecosystems by both trophic and transport linkages. PMID- 9405646 TI - A multimeric complex and the nuclear targeting of the Drosophila Rel protein Dorsal. AB - The intracellular part of the Rel signal transduction pathway in Drosophila is encoded by Toll, tube, pelle, dorsal, and cactus, and it functions to form the dorsal-ventral axis in the Drosophila embryo. Upon activation of the transmembrane receptor Toll, Dorsal dissociates from its cytoplasmic inhibitor Cactus and enters the nucleus. Tube and Pelle are required to relay the signal from Toll to the Dorsal-Cactus complex. In a yeast two-hybrid assay, we found that both Tube and Pelle interact with Dorsal. We confirmed these interactions in an in vitro binding assay. Tube interacts with Dorsal via its C-terminal domain, whereas full-length Pelle is required for Dorsal binding. Tube and Pelle bind Dorsal in the N-terminal domain 1 of the Dorsal Rel homology region rather than at the Cactus binding site. Domain 1 has been found to be necessary for Dorsal nuclear targeting. Genetic experiments indicate that Tube-Dorsal interaction is necessary for normal signal transduction. We propose a model in which Tube, Pelle, Cactus, and Dorsal form a multimeric complex that represents an essential aspect of signal transduction. PMID- 9405648 TI - Evolutionary instability of the major histocompatibility complex class I loci in New World primates. AB - Homologues of the human major histocompatibility complex (MHC) HLA-A, -B, -E, -F, and -G loci are present in all the Catarrhini (Old World primates, apes, and humans), and some of their allelic lineages have survived several speciation events. Analysis of 26 MHC class I cDNAs from seven different genera of New World primates revealed that the Callitrichinae (tamarins and marmosets) are an exception to these rules of MHC stability. In gene trees of primate MHC class I genes, sequences from the Callitrichinae cluster in a genus-specific fashion, whereas in the other genera of New World primates, as in the Catarrhini, they cluster in a transgeneric way. The genus-specific clustering of the Callitrichinae cDNAs indicates that there is no orthology between MHC class I loci in genera of this phyletic group. Additionally, the Callitrichinae genera exhibit limited variability of their MHC class I genes, in contrast to the high variability displayed by all other primates. Each Callitrichinae genus, therefore, expresses its own set of MHC class I genes, suggesting that an unusually high rate of turnover of loci occurs in this subfamily. The limited variability of MHC class I genes in the Callitrichinae is likely the result of the recent origin of these loci. PMID- 9405649 TI - Asian genotypes of JC virus in Native Americans and in a Pacific Island population: markers of viral evolution and human migration. AB - The human polyomavirus JC (JCV) causes the central nervous system demyelinating disease progressive multifocal leukoencephalopathy. Previously, we showed that 40% of Caucasians in the United States excrete JCV in the urine as detected by PCR. We have now studied 68 Navaho from New Mexico, 25 Flathead from Montana, and 29 Chamorro from Guam. By using PCR amplification of a fragment of the VP1 gene, JCV DNA was detected in the urine of 45 (66%) Navaho, 14 (56%) Flathead, and 20 (69%) Chamorro. Genotyping of viral DNAs in these cohorts by cycle sequencing showed predominantly type 2 (Asian), rather than type 1 (European). Type 1 is the major type in the United States and Hungary. Type 2 can be further subdivided into 2A, 2B, and 2C. Type 2A is found in China and Japan. Type 2B is a subtype related to the East Asian type, and is now found in Europe and the United States. The large majority (56-89%) of strains excreted by Native Americans and Pacific Islanders were the type 2A subtype, consistent with the origin of these strains in Asia. These findings indicate that JCV infection of Native Americans predates contact with Europeans, and likely predates migration of Amerind ancestors across the Bering land bridge around 12,000-30,000 years ago. If JCV had already differentiated into stable modern genotypes and subtypes prior to first settlement, the origin of JCV in humans may date from 50,000 to 100,000 years ago or more. We conclude that JCV may have coevolved with the human species, and that it provides a convenient marker for human migrations in both prehistoric and modern times. PMID- 9405650 TI - In vivo gene delivery to the liver using reconstituted chylomicron remnants as a novel nonviral vector. AB - Lipoproteins are emulsion particles that consist of lipids and apolipoproteins. Their natural function is to transport lipids and/or cholesterol to different tissues. We have taken advantage of the hydrophobic interior of these natural emulsions to solubilize DNA. Negatively charged DNA was first complexed with cationic lipids containing a quaternary amine head group. The resulting hydrophobic complex was extracted by chloroform and then incorporated into reconstituted chylomicron remnant particles ( approximately 100 nm in diameter) with an efficiency approximately 65%. When injected into the portal vein of mice, there were approximately 5 ng of a transgene product (luciferase) produced per mg of liver protein per 100 microg injected DNA. This level of transgene expression was approximately 100-fold higher than that of mice injected with naked DNA. However, such a high expression was not found after tail vein injection. Histochemical examination revealed that a large number of parenchymal cells and other types of cells in the liver expressed the transgene. Gene expression in the liver increased with increasing injected dose, and was nearly saturated with 50 microg DNA. At this dose, the expression was kept at high level in the liver for 2 days and then gradually reduced and almost disappeared by 7 days. However, by additional injection at day 7, gene expression in the liver was completely restored. By injection of plasmid DNA encoding human alpha1-antitrypsin, significant concentrations of hAAT were detected in the serum of injected animals. This is the first nonviral vector that resembles a natural lipoprotein carrier. PMID- 9405651 TI - Meis1 and pKnox1 bind DNA cooperatively with Pbx1 utilizing an interaction surface disrupted in oncoprotein E2a-Pbx1. AB - E2a-Pbx1 is a chimeric transcription factor oncoprotein produced by the t(1;19) translocation in human pre-B cell leukemia. Class I Hox proteins bind DNA cooperatively with both Pbx proteins and oncoprotein E2a-Pbx1, suggesting that leukemogenesis by E2a-Pbx1 and Hox proteins may alter transcription of cellular genes regulated by Pbx-Hox motifs. Likewise, in murine myeloid leukemia, transcriptional coactivation of Meis1 with HoxA7/A9 suggests that Meis1-HoxA7/9 heterodimers may evoke aberrant gene transcription. Here, we demonstrate that both Meis1 and its relative, pKnox1, dimerize with Pbx1 on the same TGATTGAC motif selected by dimers of Pbx proteins and unidentified partner(s) in nuclear extracts, including those from t(1;19) pre-B cells. Outside their homeodomains, Meis1 and pKnox1 were highly conserved only in two motifs required for cooperativity with Pbx1. Like the unidentified endogenous partner(s), both Meis1 and pKnox1 failed to dimerize significantly with E2a-Pbx1. The Meis1/pKnox1 interaction domain in Pbx1 resided predominantly in a conserved N-terminal Pbx domain deleted in E2a-Pbx1. Thus, the leukemic potential of E2a-Pbx1 may require abrogation of its interaction with members of the Meis and pKnox families of transcription factors, permitting selective targeting of genes regulated by Pbx Hox complexes. In addition, because most motifs bound by Pbx-Meis1/pKnox1 were not bound by Pbx1-Hox complexes, the leukemic potential of Meis1 in myeloid leukemias may involve shifting Pbx proteins from promoters containing Pbx-Hox motifs to those containing Pbx-Meis motifs. PMID- 9405652 TI - Spatio-temporally controlled site-specific somatic mutagenesis in the mouse. AB - The efficient introduction of somatic mutations in a given gene, at a given time, in a specific cell type will facilitate studies of gene function and the generation of animal models for human diseases. We have shown previously that conditional recombination-excision between two loxP sites can be achieved in mice by using the Cre recombinase fused to a mutated ligand binding domain of the human estrogen receptor (Cre-ERT), which binds tamoxifen but not estrogens. DNA excision was induced in a number of tissues after administration of tamoxifen to transgenic mice expressing Cre-ERT under the control of the cytomegalovirus promoter. However, the efficiency of excision varied between tissues, and the highest level ( approximately 40%) was obtained in the skin. To determine the efficiency of excision mediated by Cre-ERT in a given cell type, we have now crossed Cre-ERT-expressing mice with reporter mice in which expression of Escherichia coli beta-galactosidase can be induced through Cre-mediated recombination. The efficiency and kinetics of this recombination were analyzed at the cellular level in the epidermis of 6- to 8-week-old double transgenic mice. We show that site-specific excision occurred within a few days of tamoxifen treatment in essentially all epidermis cells expressing Cre-ERT. These results indicate that cell-specific expression of Cre-ERT in transgenic mice can be used for efficient tamoxifen-dependent, Cre-mediated recombination at loci containing loxP sites to generate site-specific somatic mutations in a spatio-temporally controlled manner. PMID- 9405653 TI - In vitro reconstruction of the Aspergillus (= Emericella) nidulans genome. AB - A physical map of the 31-megabase Aspergillus nidulans genome is reported, in which 94% of 5,134 cosmids are assigned to 49 contiguous segments. The physical map is the result of a two-way ordering process, in which clones and probes were ordered simultaneously on a binary DNA/DNA hybridization matrix. Compression by elimination of redundant clones resulted in a minimal map, which is a chromosome walk. Repetitive DNA is nonrandomly dispersed in the A. nidulans genome, reminiscent of heterochromatic banding patterns of higher eukaryotes. We hypothesize gene clusters may arise by horizontal transfer and spread by transposition to explain the nonrandom pattern of repeats along chromosomes. PMID- 9405654 TI - Plasmon analyses of Triticum (wheat) and Aegilops: PCR-single-strand conformational polymorphism (PCR-SSCP) analyses of organellar DNAs. AB - To investigate phylogenetic relationships among plasmons in Triticum and Aegilops, PCR-single-strand conformational polymorphism (PCR-SSCP) analyses were made of 14.0-kb chloroplast (ct) and 13. 7-kb mitochondrial (mt)DNA regions that were isolated from 46 alloplasmic wheat lines and one euplasmic line. These plasmons represent 31 species of the two genera. The ct and mtDNA regions included 10 and 9 structural genes, respectively. A total of 177 bands were detected, of which 40.6% were variable. The proportion of variable bands in ctDNA (51.1%) was higher than that of mtDNA (28. 9%). The phylogenetic trees of plasmons, derived by two different models, indicate a common picture of plasmon divergence in the two genera and suggest three major groups of plasmons (Einkorn, Triticum, and Aegilops). Because of uniparental plasmon transmission, the maternal parents of all but one polyploid species were identified. Only one Aegilops species, Ae. speltoides, was included in the Triticum group, suggesting that this species is the plasmon and B and G genome donor of all polyploid wheats. ctDNA variations were more intimately correlated with vegetative characters, whereas mtDNA variations were more closely correlated with reproductive characters. Plasmon divergence among the diploids of the two genera largely paralleled genome divergence. The relative times of origin of the polyploid species were inferred from genetic distances from their putative maternal parents. PMID- 9405655 TI - Genomic and cDNA sequence analysis of the cell matrix adhesion regulator gene. AB - The cell matrix adhesion regulator (CMAR) gene has been suggested to be a signal transduction molecule influencing cell adhesion to collagen and, through this, possibly involved in tumor suppression. The originally reported CMAR cDNA was 464 bp long with a tyrosine phosphorylation site at the extreme 3' end, which mutagenesis studies had shown to be central to the function of this gene. Since the discovery of a 4-bp insertion polymorphism within the originally reported coding region, further sequence information has been obtained. The cDNA has been extended 5' by approximately 2 kb revealing a 559-bp region showing strong homology to the proposed 5' untranslated sequence of a murine protein kinase receptor family member, variant in kinase (vik). CMAR genomic sequencing has shown the presence of an intron, the intron/exon boundary lying within this region of homology. An RNA transcript for CMAR of approximately 2.5 kb has also been identified. The data suggest complex mechanisms for control of expression of two closely associated genes, CMAR and the vik- associated sequence. PMID- 9405656 TI - Sequence of the FRA3B common fragile region: implications for the mechanism of FHIT deletion. AB - The hypothesis that chromosomal fragile sites may be "weak links" that result in hot spots for cancer-specific chromosome rearrangements was supported by the discovery that numerous cancer cell homozygous deletions and a familial translocation map within the FHIT gene, which encompasses the common fragile site, FRA3B. Sequence analysis of 276 kb of the FRA3B/FHIT locus and 22 associated cancer cell deletion endpoints shows that this locus is a frequent target of homologous recombination between long interspersed nuclear element sequences resulting in FHIT gene internal deletions, probably as a result of carcinogen-induced damage at FRA3B fragile sites. PMID- 9405657 TI - Loss of p21 increases sensitivity to ionizing radiation and delays the onset of lymphoma in atm-deficient mice. AB - Ataxia telangiectasia (AT) is an autosomal recessive disorder characterized by growth retardation, cerebellar ataxia, oculocutaneous telangiectasias, and a high incidence of lymphomas and leukemias. In addition, AT patients are sensitive to ionizing radiation. Atm-deficient mice recapitulate most of the AT phenotype. p21(cip1/waf1 )(p21 hereafter), an inhibitor of cyclin-dependent kinases, has been implicated in cellular senescence and response to gamma-radiation-induced DNA damage. To study the role of p21 in ATM-mediated signal transduction pathways, we examined the combined effect of the genetic loss of atm and p21 on growth control, radiation sensitivity, and tumorigenesis. As might have been expected, our data provide evidence that p21 modifies the in vitro senescent response seen in AT fibroblasts. Further, it is a downstream effector of ATM mediated growth control. In addition, however, we find that loss of p21 in the context of an atm-deficient mouse leads to a delay in thymic lymphomagenesis and an increase in acute radiation sensitivity in vivo (the latter principally because of effects on the gut epithelium). Modification of these two crucial aspects of the ATM phenotype can be related to an apparent increase in spontaneous apoptosis seen in tumor cells and in the irradiated intestinal epithelium of mice doubly null for atm and p21. Thus, loss of p21 seems to contribute to tumor suppression by a mechanism that operates via a sensitized apoptotic response. These results have implications for cancer therapy in general and AT patients in particular. PMID- 9405658 TI - The restriction-modification genes of Escherichia coli K-12 may not be selfish: they do not resist loss and are readily replaced by alleles conferring different specificities. AB - Type II restriction and modification (R-M) genes have been described as selfish because they have been shown to impose selection for the maintenance of the plasmid that encodes them. In our experiments, the type I R-M system EcoKI does not behave in the same way. The genes specifying EcoKI are, however, normally residents of the chromosome and therefore our analyses were extended to monitor the deletion of chromosomal genes rather than loss of plasmid vector. If EcoKI were to behave in the same way as the plasmid-encoded type II R-M systems, the loss of the relevant chromosomal genes by mutation or recombination should lead to cell death because the cell would become deficient in modification enzyme and the bacterial chromosome would be vulnerable to the restriction endonuclease. Our data contradict this prediction; they reveal that functional type I R-M genes in the chromosome are readily replaced by mutant alleles and by alleles encoding a type I R-M system of different specificity. The acquisition of allelic genes conferring a new sequence specificity, but not the loss of the resident genes, is dependent on the product of an unlinked gene, one predicted [Prakash-Cheng, A., Chung, S. S. & Ryu, J. (1993) Mol. Gen. Genet. 241, 491-496] to be relevant to control of expression of the genes that encode EcoKI. Our evidence suggests that not all R-M systems are evolving as "selfish" units; rather, the diversity and distribution of the family of type I enzymes we have investigated require an alternative selective pressure. PMID- 9405659 TI - Protamine-Cre recombinase transgenes efficiently recombine target sequences in the male germ line of mice, but not in embryonic stem cells. AB - The production of subtle or conditional mutations in mice through the combined use of site-specific and homologous recombination has become an increasingly widespread experimental paradigm in mammalian genetics. Embryonic stem cells containing recombinase transgenes that were expressed in the male germ line, but not in other tissues or in the embryonic stem cells themselves, would substantially simplify the production of such alleles. Here we show that transgenes comprised of the mouse protamine 1 promoter and the Cre recombinase coding sequence mediate the efficient recombination of a Cre target transgene in the male germ line, but not in other tissues. Embryonic stem cell lines generated from one of these transgenic strains were transfected with targeting vectors that included loxP-flanked selectable markers, and homologously recombined alleles containing the marker and functional loxP sites were isolated. These results establish the potential of the system for substantially reducing the time, effort, and resources required to produce homologously recombined alleles in mice that have been secondarily rearranged by a site-specific recombinase. PMID- 9405660 TI - Comparative mapping of the human 22q11 chromosomal region and the orthologous region in mice reveals complex changes in gene organization. AB - The region of human chromosome 22q11 is prone to rearrangements. The resulting chromosomal abnormalities are involved in Velo-cardio-facial and DiGeorge syndromes (VCFS and DGS) (deletions), "cat eye" syndrome (duplications), and certain types of tumors (translocations). As a prelude to the development of mouse models for VCFS/DGS by generating targeted deletions in the mouse genome, we examined the organization of genes from human chromosome 22q11 in the mouse. Using genetic linkage analysis and detailed physical mapping, we show that genes from a relatively small region of human 22q11 are distributed on three mouse chromosomes (MMU6, MMU10, and MMU16). Furthermore, although the region corresponding to about 2.5 megabases of the VCFS/DGS critical region is located on mouse chromosome 16, the relative organization of the region is quite different from that in humans. Our results show that the instability of the 22q11 region is not restricted to humans but may have been present throughout evolution. The results also underscore the importance of detailed comparative mapping of genes in mice and humans as a prerequisite for the development of mouse models of human diseases involving chromosomal rearrangements. PMID- 9405661 TI - Drosophila host defense: differential induction of antimicrobial peptide genes after infection by various classes of microorganisms. AB - Insects respond to microbial infection by the rapid and transient expression of several genes encoding potent antimicrobial peptides. Herein we demonstrate that this antimicrobial response of Drosophila is not aspecific but can discriminate between various classes of microorganisms. We first observe that the genes encoding antibacterial and antifungal peptides are differentially expressed after injection of distinct microorganisms. More strikingly, Drosophila that are naturally infected by entomopathogenic fungi exhibit an adapted response by producing only peptides with antifungal activities. This response is mediated through the selective activation of the Toll pathway. PMID- 9405662 TI - Identification of functional domains on human interleukin 10. AB - Interleukin 10 (IL-10) is a recently described natural endogenous immunosuppressive cytokine that has been identified in human, murine, and other organisms. Human IL-10 (hIL-10) has high homology with murine IL-10 (mIL-10) as well as with an Epstein-Barr virus genome product BCRFI. This viral IL-10 (vIL 10) shares a number of activities with hIL-10. IL-10 significantly affects chemokine biology, because human IL-10 inhibits chemokine production and is a specific chemotactic factor for CD8+ T cells. It suppresses the ability of CD4+ T cells, but not CD8+ T cells, to migrate in response to IL-8. A nonapeptide (IT9302) with complete homology to a sequence of hIL-10 located in the C-terminal portion (residues 152-160) of the cytokine was found to possess activities that mimic some of those of hIL-10. These are: (i) inhibition of IL-1beta-induced IL-8 production by peripheral blood mononuclear cell, (ii) inhibition of spontaneous IL-8 production by cultured human monocytes, (iii) induction of IL-1 receptor antagonistic protein production by human monocytes, (iv) induction of chemotactic migration of CD8+ human T lymphocytes in vitro, (v) desensitization of human CD8+ T cells resulting in an unresponsiveness toward rhIL-10-induced chemotaxis, (vi) suppression of the chemotactic response of CD4+ T human lymphocytes toward IL-8, (vii) induction of IL-4 production by cultured normal human CD4+ T cells, (viii) down-regulation of tumor necrosis factor-alpha production by CD8+ T cells, and (ix) inhibition of class II major histocompatibility complex antigen expression on IFN-gamma-stimulated human monocytes. Another nonapeptide (IT9403) close to the NH2-terminal part of hIL-10 did not reveal cytokine synthesis inhibitory properties, but proved to be a regulator of mast cell proliferation. In conclusion, we have identified two functional domains of IL-10 exerting different IL-10 like activities, an observation that suggests that relatively small segments of these signal proteins are responsible for particular biological functions. PMID- 9405663 TI - DNA immunization: induction of higher avidity antibody and effect of route on T cell cytotoxicity. AB - Immunizations of mice with plasmid DNAs encoding ovalbumin (OVA), human Ig, and hen egg lysozyme were compared with doses of soluble protein (without adjuvant) that induced similar IgG responses. The route of immunization influenced the magnitude of the antibody (Ab) response in that intradermal (i.d.) injection elicited higher IgG Ab levels than i.m. injection in both DNA- and protein immunized mice. Although total IgG levels were similar to soluble protein controls, the avidity of the anti-OVA Abs generated by DNA immunization were 100- and 1,000-fold higher via the i.m. or i.d. route, respectively. However, despite the generation of high-avidity Ab in DNA-immunized mice, germinal centers could not be detected in either DNA- or protein-immunized mice. Examination of the IgG subclass response showed that IgG2a was induced by i.m. DNA immunization, coinciding with elevated interferon gamma production, whereas a dominant and elevated IgG1 response, coinciding with detectable interleukin 4 production, was generated after i.d. immunization with DNA or soluble OVA and hen egg lysozyme but not human Ig protein. As expected, cytotoxic T cell (CTL) responses could be detected only after DNA immunization. I.d. immunization produced the strongest CTL responses early (2 weeks) but was similar to i.m. later. Therefore, DNA immunization can differ from protein immunization by its ability to induce rapid CTL responses and higher avidity Ab, both of which are advantageous for vaccination. PMID- 9405664 TI - Purified hematopoietic stem cells without facilitating cells can repopulate fully allogeneic recipients across entire major histocompatibility complex transplantation barrier in mice. AB - We report herein the successful long term engraftment of highly purified hematopoietic stem cells (HSCs) without any facilitating cells in fully allogeneic recipient mice across the entire major histocompatibility complex (MHC) transplantation barrier. This finding challenges the assumption that highly purified marrow HSCs alone cannot produce long-lived allogeneic bone marrow chimeras across the MHC barrier. In the present experiments, 1 x 10(5) HSCs from 5-fluorouracil (5-FU)-treated donors, without any facilitating cells, have been found to repopulate lethally irradiated fully allogeneic recipients. Low density, lineage-negative (CD4-, CD8-, B220-, Mac-1-, Gr-1-), CD71-negative, class I highly positive, FACS-sorted cells from 5-FU-treated C57BL/6 (B6) donor mice were transplanted into lethally irradiated BALB/c recipients. (BALB/c --> BALB/c) --> BALB/c T cell-depleted marrow cells used as compromised cells were also transplanted into the recipients to permit experiments to be pursued over a long period of time. Cells of donor origin in all recognized lineages of hematopoietic cells developed in these allogeneic chimeras. One thousand HSCs were sufficient to repopulate hemiallogeneic recipients, but 1 x 10(4) HSCs alone from 5-FU treated donors failed to repopulate the fully allogeneic recipients. Transplantation of primary marrow stromal cells or bones of the donor strain into recipient, together with 1 x 10(4) HSCs, also failed to reconstitute fully allogeneic recipients. Suppression of resistance of recipients by thymectomy or injections of granulocyte colony-stimulating factor before stem cell transplantation enhanced the engraftment of allogeneic HSCs. Our experiments show that reconstitution of all lymphohematopoietic lineages across the entire MHC transplantation barriers may be achieved by transplanting allogeneic HSCs alone, without any facilitating cells, as long as a sufficient number of HSCs is transplanted. PMID- 9405665 TI - Mice transgenic for human CD4 and CCR5 are susceptible to HIV infection. AB - HIV entry into human cells is mediated by CD4 acting in concert with one of several members of the chemokine receptor superfamily. The resistance to HIV infection observed in individuals with defective CCR5 alleles indicated that this particular chemokine receptor plays a crucial role in the initiation of in vivo HIV infection. Expression of human CD4 transgene does not render mice susceptible to HIV infection because of structural differences between human and mouse CCR5. To ascertain whether expression of human CD4 and CCR5 is sufficient to make murine T lymphocytes susceptible to HIV infection, the lck promoter was used to direct the T cell-specific expression of human CD4 and CCR5 in transgenic mice. Peripheral blood mononuclear cells and splenocytes isolated from these mice expressed human CD4 and CCR5 and were infectible with selected M-tropic HIV isolates. After in vivo inoculation, HIV-infected cells were detected by DNA PCR in the spleen and lymph nodes of these transgenic mice, but HIV could not be cultured from these cells. This indicated that although transgenic expression of human CD4 and CCR5 permitted entry of HIV into the mouse cells, significant HIV infection was prevented by other blocks to HIV replication present in mouse cells. In addition to providing in vivo verification for the important role of CCR5 in T lymphocyte HIV infection, these transgenic mice represent a new in vivo model for understanding HIV pathogenesis by delineating species-specific cellular factors required for productive in vivo HIV infection. These mice should also prove useful for the assessment of potential therapeutic and preventative modalities, particularly vaccines. PMID- 9405666 TI - Superactivation of an immune response triggered by oligomerized T cell epitopes. AB - The peptides bound to class II major histocompatibility complex (MHC) molecules extend out both ends of the peptide binding groove. This structural feature provided the opportunity to design multivalent polypeptide chains that cross-link class II MHC molecules through multiple, repetitive MHC binding sites. By using recombinant techniques, polypeptide oligomers were constructed that consist of up to 32 copies of an HLA-DR1-restricted T cell epitope. The epitope HA306-318, derived from influenza virus hemagglutinin, was connected by 12- to 36-aa long spacer sequences. These oligomers were found to cross-link soluble HLA-DR1 molecules efficiently and, upon binding to the MHC molecules of a monocyte line, to trigger signal transduction indicated by the enhanced expression of some cell surface molecules. A particularly strong effect was evident in the T cell response. A hemagglutinin-specific T cell clone recognized these antigens at concentrations up to three to four orders of magnitude lower than that of the peptide or the hemagglutinin protein. Both signal transduction in the monocyte and the proliferative response of the T cell were affected greatly by the length of the oligomer (i.e., the number of repetitive units) and the distance of the epitopes within the oligomer (spacing). Thus, the formation of defined clusters of T cell receptor/MHC/peptide antigen complexes appears to be crucial for triggering the immune response and can be used to enhance the antigenicity of a peptide antigen by oligomerizing the epitope. PMID- 9405668 TI - Role of the major histocompatibility complex class II Ea gene in lupus susceptibility in mice. AB - The gene(s) encoded within major histocompatibility complex (MHC) act as one of the major genetic elements contributing to the susceptibility of murine systemic lupus erythematosus (SLE). We have recently demonstrated that lupus susceptibility is more closely linked to the I-E- H-2(b) haplotype than to the I E+ H-2(d) haplotype in lupus-prone BXSB and (NZB x BXSB)F1 hybrid mice. To investigate whether the reduced susceptibility to SLE in H-2(d) mice is related to the expression of the MHC class II Ea gene (absent in H-2(b) mice), we determined the possible role of the Ea gene as a lupus protective gene in mice. Our results showed that (i) the development of SLE was almost completely prevented in BXSB (H-2(b)) mice expressing two copies of the Ead transgene at the homozygous level as well as in BXSB H-2(k) (I-E+) congenic mice as for H-2(d) BXSB mice, and (ii) the expression of two functional Ea (transgenic and endogenous) genes in either H-2(d/b) (NZB x BXSB)F1 or H-2(k/b) (MRL x BXSB)F1 mice provided protection from SLE at levels comparable to those conferred by the H-2(d/d) or H-2(k/k) haplotype. In addition, the level of the Ea gene-mediated protection appeared to be dependent on the genetic susceptibility to SLE in individual lupus-prone mice. Our results indicate that the reduced susceptibility associated with the I-E+ H-2(d) and H-2(k) haplotypes (versus the I-E- H-2(b) haplotype) is largely, if not all, contributed by the apparent autoimmune suppressive effect of the Ea gene, independently of the expression of the I-A or other MHC-linked genes. PMID- 9405667 TI - Role of B cells in antigen presentation of the hepatitis B core. AB - The hepatitis B virus (HBV) nucleocapsid or core antigen (HBcAg) is extremely immunogenic during infection and after immunization. For example, during many chronic infections, HBcAg is the only antigen capable of eliciting an immune response, and nanogram amounts of HBcAg elicit antibody production in mice. Recent structural analysis has revealed a number of characteristics that may help explain this potent immunogenicity. Our analysis of how the HBcAg is presented to the immune system revealed that the HBcAg binds to specific membrane Ig (mIg) antigen receptors on a high frequency of resting, murine B cells sufficiently to induce B7.1 and B7.2 costimulatory molecules. This enables HBcAg-specific B cells from unprimed mice to take up, process, and present HBcAg to naive Th cells in vivo and to T cell hybridomas in vitro approximately 10(5) times more efficiently than classical macrophage or dendritic antigen-presenting cells (APC). These results reveal a structure-function relation for the HBcAg, confirm that B cells can function as primary APC, explain the enhanced immunogenicity of HBcAg, and may have relevance for the induction and/or maintenance of chronic HBV infection. PMID- 9405669 TI - Protective anti-tumor immunity induced by vaccination with recombinant adenoviruses encoding multiple tumor-associated cytotoxic T lymphocyte epitopes in a string-of-beads fashion. AB - Vaccines harboring genes that encode functional oncoproteins are intrinsically hazardous, as their application may lead to introduction of these genes into normal cells and thereby to tumorigenesis. On the other hand, oncoproteins are especially attractive targets for immunotherapy of cancer, as their expression is generally required for tumor growth, making the arisal of tumor variants lacking these antigens unlikely. Using murine tumor models, we investigated the efficacy of polyepitope recombinant adenovirus (rAd) vaccines, which encode only the immunogenic T cell epitopes derived from several oncogenes, for the induction of protective anti-tumor immunity. We chose to employ rAd, as these are safe vectors that do not induce the side effects associated with, for example, vaccinia virus vaccines. A single polyepitope rAd was shown to give rise to presentation of both H-2 and human leukocyte antigen-restricted cytotoxic T lymphocyte (CTL) epitopes. Moreover, vaccination with a rAd encoding H-2-restricted CTL epitopes, derived from human adenovirus type 5 early region 1 and human papilloma virus type 16 induced tumors, elicited strong tumor-reactive CTL and protected the vaccinated animals against an otherwise lethal challenge with either of these tumors. The protection induced was superior compared with that obtained by vaccination with irradiated tumor cells. Thus, vaccination with polyepitope rAd is a powerful approach for the induction of protective anti-tumor immunity that allows simultaneous immunization against multiple tumor-associated T cell epitopes, restricted by various major histocompatibility complex haplotypes. PMID- 9405670 TI - Multiple receptors for HLA-G on human natural killer cells. AB - HLA-G is the putative natural killer (NK) cell inhibitory ligand expressed on the extravillous cytotrophoblast of the human placenta. Killing of the class I negative human B cell line 721.221 by NK cells is inhibited by the expression of HLA-G. This inhibition is dependent on a high level of HLA-G expression. In the present study, the nature of the receptors that mediate the inhibition has been studied with 140 NK cell lines from two donors and 246 NK clones from 5 donors by blocking the inhibition using monoclonal antibodies against the known NK inhibitory receptors: CD158a, CD158b, and CD94. Both CD94 and the two CD158 proteins can function as receptors, although the former clearly predominates. In many cases, a combination of antibodies to these receptors is required to achieve maximal reversal of inhibition. Moreover, in at least one-third of the NK cells that are inhibited by HLA-G, these antibodies alone or in combination do not reverse inhibition, strongly suggesting the existence of a third major unidentified receptor for HLA-G. PMID- 9405671 TI - WIP, a protein associated with wiskott-aldrich syndrome protein, induces actin polymerization and redistribution in lymphoid cells. AB - Wiskott-Aldrich syndrome (WAS) is an X-linked immunodeficiency caused by mutations that affect the WAS protein (WASP) and characterized by cytoskeletal abnormalities in hematopoietic cells. By using the yeast two-hybrid system we have identified a proline-rich WASP-interacting protein (WIP), which coimmunoprecipitated with WASP from lymphocytes. WIP binds to WASP at a site distinct from the Cdc42 binding site and has actin as well as profilin binding motifs. Expression of WIP in human B cells, but not of a WIP truncation mutant that lacks the actin binding motif, increased polymerized actin content and induced the appearance of actin-containing cerebriform projections on the cell surface. These results suggest that WIP plays a role in cortical actin assembly that may be important for lymphocyte function. PMID- 9405672 TI - Combined transgenic expression of alpha-galactosidase and alpha1,2 fucosyltransferase leads to optimal reduction in the major xenoepitope Galalpha(1,3)Gal. AB - Hyperacute rejection of pig organs by humans involves the interaction of Galalpha(1,3)Gal with antibodies and complement. Strategies to reduce the amount of xenoantigen Galalpha(1,3)Gal were investigated by overexpression of human lysosomal alpha-galactosidase in cultured porcine cells and transgenic mice. The overexpression of human alpha-galactosidase in cultured porcine endothelial cells and COS cells resulted in a 30-fold reduction of cell surface Galalpha(1,3)Gal and a 10-fold reduction in cell reactivity with natural human antibodies. Splenocytes from transgenic mice overexpressing human alpha-galactosidase showed only a 15-25% reduction in binding to natural human anti-Galalpha(1,3)Gal antibodies; however, this decrease was functionally significant as demonstrated by reduced susceptibility to human antibody-mediated lysis. However, because there is residual Galalpha(1,3)Gal and degalactosylation results in the exposure of N-acetyllactosamine residues and potential new xenoepitopes, using alpha galactosidase alone is unlikely to overcome hyperacute rejection. We previously reported that mice overexpressing human alpha1,2-fucosyltransferase as a transgene had approximately 90% reduced Galalpha(1,3)Gal levels due to masking of the xenoantigen by fucosylation; we evaluated the effect of overexpressing alpha galactosidase and alpha1,2-fucosyltransferase on Galalpha(1,3)Gal levels. Galalpha(1, 3)Gal-positive COS cells expressing alpha1,3-galactosyltransferase, alpha1,2-fucosyltransferase, and alpha-galactosidase showed negligible cell surface staining and were not susceptible to lysis by human serum containing antibody and complement. Thus, alpha1, 2-fucosyltransferase and alpha galactosidase effectively reduced the expression of Galalpha(1,3)Gal on the cell surface and could be used to produce transgenic pigs with negligible levels of cell surface Galalpha(1,3)Gal, thereby having no reactivity with human serum and improving graft survival. PMID- 9405673 TI - The anticoagulant activation of antithrombin by heparin. AB - Antithrombin, a plasma serpin, is relatively inactive as an inhibitor of the coagulation proteases until it binds to the heparan side chains that line the microvasculature. The binding specifically occurs to a core pentasaccharide present both in the heparans and in their therapeutic derivative heparin. The accompanying conformational change of antithrombin is revealed in a 2.9-A structure of a dimer of latent and active antithrombins, each in complex with the high-affinity pentasaccharide. Inhibitory activation results from a shift in the main sheet of the molecule from a partially six-stranded to a five-stranded form, with extrusion of the reactive center loop to give a more exposed orientation. There is a tilting and elongation of helix D with the formation of a 2-turn helix P between the C and D helices. Concomitant conformational changes at the heparin binding site explain both the initial tight binding of antithrombin to the heparans and the subsequent release of the antithrombin-protease complex into the circulation. The pentasaccharide binds by hydrogen bonding of its sulfates and carboxylates to Arg-129 and Lys-125 in the D-helix, to Arg-46 and Arg-47 in the A helix, to Lys-114 and Glu-113 in the P-helix, and to Lys-11 and Arg-13 in a cleft formed by the amino terminus. This clear definition of the binding site will provide a structural basis for developing heparin analogues that are more specific toward their intended target antithrombin and therefore less likely to exhibit side effects. PMID- 9405674 TI - Expression of lipocalin-type prostaglandin D synthase (beta-trace) in human heart and its accumulation in the coronary circulation of angina patients. AB - Lipocalin-type prostaglandin D synthase (L-PGDS) is localized in the central nervous system and male genital organs of various mammals and is secreted as beta trace into the closed compartment of these tissues separated from the systemic circulation. In this study, we found that the mRNA for the human enzyme was expressed most intensely in the heart among various tissues examined. In human autopsy specimens, the enzyme was localized immunocytochemically in myocardial cells, atrial endocardial cells, and a synthetic phenotype of smooth muscle cells in the arteriosclerotic intima, and accumulated in the atherosclerotic plaque of coronary arteries with severe stenosis. In patients with stable angina (75-99% stenosis), the plasma level of L-PGDS was significantly (P < 0.05) higher in the great cardiac vein (0.694 +/- 0.054 microg/ml, n = 7) than in the coronary artery (0.545 +/- 0.034 microg/ml), as determined by a sandwich enzyme immunoassay. However, the veno-arterial difference in the plasma L-PGDS concentration was not observed in normal subjects without stenosis. After a percutaneous transluminal coronary angioplasty was performed to compress the stenotic atherosclerotic plaques, the L-PGDS concentration in the cardiac vein decreased significantly (P < 0.05) to 0.610 +/- 0.051 microg/ml at 20 min and reached the arterial level within 1 h. These findings suggest that L-PGDS is present in both endocardium and myocardium of normal subjects and the stenotic site of patients with stable angina and is secreted into the coronary circulation. PMID- 9405675 TI - Development of an epithelium-specific expression cassette with human DNA regulatory elements for transgene expression in lung airways. AB - The efficient expression of therapeutic genes in target cells or tissues is an important component of efficient and safe gene therapy. Utilizing regulatory elements from the human cytokeratin 18 (K18) gene, including 5' genomic sequences and one of its introns, we have developed a novel expression cassette that can efficiently express reporter genes, as well as the human cystic fibrosis transmembrane conductance regulator (CFTR) gene, in cultured lung epithelial cells. CFTR transcripts expressed from the native K18 enhancer/promoter include two alternative splicing products, due to the activation of two cryptic splice sites in the CFTR coding region. Modification of the K18 intron and CFTR cDNA sequences eliminated the cryptic splice sites without changing the CFTR amino acid sequence, and led to enhanced CFTR mRNA and protein expression as well as biological function. Transgenic expression analysis in mice showed that the modified expression cassette can direct efficient and epithelium-specific expression of the Escherichia coli LacZ gene in the airways of fetal lungs, with no detectable expression in lung fibroblasts or endothelial cells. This is the first expression cassette which selectively directs lung transgene expression for CFTR gene therapy to airway epithelia. PMID- 9405677 TI - Hepatitis B virus X protein and p53 tumor suppressor interactions in the modulation of apoptosis. AB - We have reported previously that the hepatitis B virus oncoprotein, HBx, can bind to the C terminus of p53 and inhibit several critical p53-mediated cellular processes, including DNA sequence-specific binding, transcriptional transactivation, and apoptosis. Recognizing the importance of p53-mediated apoptosis for maintaining homeostasis and preventing neoplastic transformation, here we further examine the physical interaction between HBx and p53 as well as the functional consequences of this association. In vitro binding studies indicate that the ayw and adr viral subtypes of HBx bind similar amounts of glutathione S-transferase-p53 with the distal C terminus of HBx (from residues 111 to 154) being critical for this interaction. Using a microinjection technique, we show that this same C-terminal region of HBx is necessary for sequestering p53 in the cytoplasm and abrogating p53-mediated apoptosis. The transcriptional transactivation domain of HBx also maps to its C terminus; however, a comparison of the ability of full-length and truncated HBx protein to abrogate p53-induced apoptosis versus transactivate simian virus 40- or human nitric oxide synthase-2 promoter-driven reporter constructs indicates that these two functional properties are distinct and thus may contribute to hepatocarcinogenesis differently. Collectively, our data indicate that the distal C-terminal domain of HBx, independent of its transactivation activity, complexes with p53 in the cytoplasm, partially preventing its nuclear entry and ability to induce apoptosis. These pathobiological effects of HBx may contribute to the early stages of hepatocellular carcinogenesis. PMID- 9405676 TI - Regulated expression of the diphtheria toxin A chain by a tumor-specific chimeric transcription factor results in selective toxicity for alveolar rhabdomyosarcoma cells. AB - Alveolar rhabdomyosarcoma (ARMS) cells often harbor one of two unique chromosomal translocations, either t(2;13)(q35;q14) or t(1;13)(p36;q14). The chimeric proteins expressed from these rearrangements, PAX3-FKHR and PAX7-FKHR, respectively, are potent transcriptional activators. In an effort to exploit these unique cancer-specific molecules to achieve ARMS-specific expression of therapeutic genes, we have studied the expression of a minimal promoter linked to six copies of a PAX3 DNA binding site, prs-9. In transient transfections, expression of the prs-9-regulated reporter genes was approximately 250-fold higher than expression of genes lacking the prs-9 sequences in cell lines derived from ARMS, but remained at or below baseline levels in other cells. High expression of these prs-9-regulated genes was also observed in a cancer cell line that lacks t(2;13) but was stably transfected with a plasmid expressing PAX3 FKHR. Transfection of a plasmid containing the diphtheria toxin A chain gene regulated by prs-9 sequences (pA3-6PED) was selectively cytotoxic for PAX3-FKHR expressing cells. This was shown by inhibition of gene expression from cotransfected plasmids and by direct cytotoxicity after transfected cells were isolated by cell sorting. Gene transfer of pA3-6PED may thus be useful as a cancer-specific treatment strategy for t(2;13)- or t(1;13)-positive ARMS. Furthermore, gene transfer of fusion protein-regulated toxin genes might also be applied to the treatment of other cancers that harbor cancer-specific chromosomal translocations involving transcription factors. PMID- 9405678 TI - Identification of semaphorin E as a non-MDR drug resistance gene of human cancers. AB - To improve cancer chemotherapy, a better understanding of the molecular mechanisms of drug resistance is essential. To identify the molecules responsible for drug resistance that is unrelated to MDR1 or MRP gene products, a eukaryotic expression cDNA library of cis-diamminedichloroplatinum(II) (CDDP)-resistant ovarian cancer TYKnuR cells was introduced into Cos-7 cells. After repeated CDDP selection, cDNA homologous to murine semaphorin E was isolated from surviving cells. Human semaphorin E (H-sema E) was overexpressed in CDDP-resistant cell lines and was readily induced not only by diverse chemotherapeutic drugs but also by x-ray and UV irradiation. Transfection of H-sema E conferred a drug-resistant phenotype to CDDP-sensitive cells. In addition, the aberrant expression of H-sema E protein was detected immunohistochemically in 14 of 42 (33.3%) recurrent squamous cell carcinomas removed at autopsy after extensive radiochemotherapy. Recently, another member of the semaphorin family, CD100, was shown to significantly improve the viability of B lymphocytes. These results suggest the involvement of semaphorins in diverse cell survival mechanisms. PMID- 9405679 TI - Analysis of genomic alterations in benign, atypical, and anaplastic meningiomas: toward a genetic model of meningioma progression. AB - Nineteen benign [World Health Organization (WHO) grade I; MI], 21 atypical (WHO grade II; MII), and 19 anaplastic (WHO grade III; MIII) sporadic meningiomas were screened for chromosomal imbalances by comparative genomic hybridization (CGH). These data were supplemented by molecular genetic analyses of selected chromosomal regions and genes. With increasing malignancy grade, a marked accumulation of genomic aberrations was observed; i.e., the numbers (mean +/- SEM) of total alterations detected per tumor were 2.9 +/- 0.7 for MI, 9.2 +/- 1.2 for MII, and 13.3 +/- 1.9 for MIII. The most frequent alteration detected in MI was loss on 22q (58%). In MII, aberrations most commonly identified were losses on 1p (76%), 22q (71%), 14q (43%), 18q (43%), 10 (38%), and 6q (33%), as well as gains on 20q (48%), 12q (43%), 15q (43%), 1q (33%), 9q (33%), and 17q (33%). In MIII, most of these alterations were found at similar frequencies. However, an increase in losses on 6q (53%), 10 (68%), and 14q (63%) was observed. In addition, 32% of MIII demonstrated loss on 9p. Homozygous deletions in the CDKN2A gene at 9p21 were found in 4 of 16 MIII (25%). Highly amplified DNA sequences were mapped to 12q13-q15 by CGH in 1 MII. Southern blot analysis of this tumor revealed amplification of CDK4 and MDM2. By CGH, DNA sequences from 17q were found to be amplified in 1 MII and 8 MIII, involving 17q23 in all cases. Despite the high frequency of chromosomal aberrations in the MII and MIII investigated, none of these tumors showed mutations in exons 5-8 of the TP53 gene. On the basis of the most common aberrations identified in the various malignancy grades, a model for the genomic alterations associated with meningioma progression is proposed. PMID- 9405680 TI - In vivo NMR and MRI using injection delivery of laser-polarized xenon. AB - Because xenon NMR is highly sensitive to the local environment, laser-polarized xenon could be a unique probe of living tissues. Realization of clinical and medical science applications beyond lung airspace imaging requires methods of efficient delivery of laser-polarized xenon to tissues, because of the short spin lattice relaxation times and relatively low concentrations of xenon attainable in the body. Preliminary results from the application of a polarized xenon injection technique for in vivo 129Xe NMR/MRI are extrapolated along with a simple model of xenon transit to show that the peak local concentration of polarized xenon delivered to tissues by injection may exceed that delivered by respiration by severalfold. PMID- 9405681 TI - Hypertension, cardiac hypertrophy, and sudden death in mice lacking natriuretic peptide receptor A. AB - Natriuretic peptides, produced in the heart, bind to the natriuretic peptide receptor A (NPRA) and cause vasodilation and natriuresis important in the regulation of blood pressure. We here report that mice lacking a functional Npr1 gene coding for NPRA have elevated blood pressures and hearts exhibiting marked hypertrophy with interstitial fibrosis resembling that seen in human hypertensive heart disease. Echocardiographic evaluation of the mice demonstrated a compensated state of systemic hypertension in which cardiac hypertrophy and dilatation are evident but with no reduction in ventricular performance. Nevertheless, sudden death, with morphologic evidence indicative in some animals of congestive heart failure and in others of aortic dissection, occurred in all 15 male mice lacking Npr1 before 6 months of age, and in one of 16 females in our study. Thus complete absence of NPRA causes hypertension in mice and leads to cardiac hypertrophy and, particularly in males, lethal vascular events similar to those seen in untreated human hypertensive patients. PMID- 9405682 TI - alpha+-Thalassemia protects children against disease caused by other infections as well as malaria. AB - In the South West Pacific region, the striking geographical correlation between the frequency of alpha+-thalassemia and the endemicity of Plasmodium falciparum suggests that this hemoglobinopathy provides a selective advantage against malaria. In Vanuatu, paradoxically, alpha+-thalassemia increases the incidence of contracting mild malaria in the first 2 years of life, but severe disease was too uncommon to assess adequately. Therefore, we undertook a prospective case-control study of children with severe malaria on the north coast of Papua New Guinea, where malaria transmission is intense and alpha+-thalassemia affects more than 90% of the population. Compared with normal children, the risk of having severe malaria was 0.40 (95% confidence interval 0.22-0.74) in alpha+-thalassemia homozygotes and 0.66 (0.37-1.20) in heterozygotes. Unexpectedly, the risk of hospital admission with infections other than malaria also was reduced to a similar degree in homozygous (0. 36; 95% confidence interval 0.22-0.60) and heterozygous (0.63; 0. 38-1.07) children. This clinical study demonstrates that a malaria resistance gene protects against disease caused by infections other than malaria. The mechanism of the remarkable protective effect of alpha+-thalassemia against severe childhood disease remains unclear but must encompass the clear interaction between this hemoglobinopathy and both malarial and nonmalarial infections. PMID- 9405683 TI - CD4-independent, CCR5-dependent infection of brain capillary endothelial cells by a neurovirulent simian immunodeficiency virus strain. AB - Brain capillary endothelial cells (BCECs) are targets of CD4-independent infection by HIV-1 and simian immunodeficiency virus (SIV) strains in vitro and in vivo. Infection of BCECs may provide a portal of entry for the virus into the central nervous system and could disrupt blood-brain barrier function, contributing to the development of AIDS dementia. We found that rhesus macaque BCECs express chemokine receptors involved in HIV and SIV entry including CCR5, CCR3, CXCR4, and STRL33, but not CCR2b, GPR1, or GPR15. Infection of BCECs by the neurovirulent strain SIV/17E-Fr was completely inhibited by aminooxypentane regulation upon activation, normal T cell expression and secretion in the presence or absence of ligands, but not by eotaxin or antibodies to CD4. We found that the envelope (env) proteins from SIV/17E-Fr and several additional SIV strains mediated cell-cell fusion and virus infection with CD4-negative, CCR5 positive cells. In contrast, fusion with cells expressing the coreceptors STRL33, GPR1, and GPR15 was CD4-dependent. These results show that CCR5 can serve as a primary receptor for SIV in BCECs and suggest a possible CD4-independent mechanism for blood-brain barrier disruption and viral entry into the central nervous system. PMID- 9405684 TI - Genetically determined chloride-sensitive hypertension and stroke. AB - The stroke-prone spontaneously hypertensive rat (SHRSP) is a genetically determined model of "salt-sensitive" stroke and hypertension whose full phenotypic expression is said to require a diet high in Na+ and low in K+. We tested the hypothesis that dietary Cl- determines the phenotypic expression of the SHRSP. In the SHRSP fed a normal NaCl diet, supplementing dietary K+ with KCl exacerbated hypertension, whereas supplementing either KHCO3 or potassium citrate (KB/C) attenuated hypertension, when blood pressure (BP) was measured radiotelemetrically, directly and continually. Supplemental KCl, but not KB/C, induced strokes, which occurred in all and only those rats in the highest quartiles of both BP and plasma renin activity (PRA). PRA was higher with KCl than with KB/C. These observations demonstrate that with respect to both severity of hypertension and frequency of stroke the phenotypic expression of the SHRSP is (i) either increased or decreased, depending on whether the anionic component of the potassium salt supplemented is, or is not, Cl-; (ii) increased by supplementing Cl- without supplementing Na+, and despite supplementing K+; and hence (iii) both selectively Cl--sensitive and Cl--determined. The observations suggest that in the SHRSP selectively supplemented with Cl- the likelihood of stroke depends on the extent to which both BP and PRA increase. PMID- 9405685 TI - p53CP, a putative p53 competing protein that specifically binds to the consensus p53 DNA binding sites: a third member of the p53 family? AB - p53 tumor suppressor protein negatively regulates cell growth, mainly through the transactivation of its downstream target genes. As a sequence-specific DNA binding transcription factor, p53 specifically binds to a 20-bp consensus motif 5'-PuPuPuC(A/T) (T/A)GPyPyPyPuPuPuC(A/T)(T/A)GPyPyPy-3'. We have now identified, partially purified, and characterized an additional approximately 40-kDa nuclear protein, p53CP (p53 competing protein), that specifically binds to the consensus p53 binding sites found in several p53 downstream target genes, including Waf-1, Gadd45, Mdm2, Bax, and RGC. The minimal sequence requirement for binding is a 14 bp motif, 5'-CTTGCTTGAACAGG-3' [5'-C(A/T)(T/A)GPyPyPyPuPuPuC(A/T)(T/A)G-3'], which includes the central nucleotides of the typical p53 binding site with one mismatch. p53CP and p53 (complexed with antibody) showed a similar binding specificity to Waf-1 site but differences in Gadd45 and T3SF binding. Like p53, p53CP also binds both double- and single-stranded DNA oligonucleotides. Important to note, cell cycle blockers and DNA damaging reagents, which induce p53 binding activity, were found to inhibit p53CP binding in p53-positive, but not in p53 negative, cells. This finding suggested a p53-dependent coordinate regulation of p53 and p53CP in response to external stimuli. p53CP therefore could be a third member of the p53 family, in addition to p53 and p73, a newly identified p53 homolog. p53CP, if sequestering p53 from its DNA binding sites through competitive binding, may provide a novel mechanism of p53 inactivation. Alternatively, p53CP may have p53-like functions by binding and transactivating p53 downstream target genes. Cloning of the p53CP gene ultimately will resolve this issue. PMID- 9405686 TI - Cloning and mutagenesis of a herpesvirus genome as an infectious bacterial artificial chromosome. AB - A strategy for cloning and mutagenesis of an infectious herpesvirus genome is described. The mouse cytomegalovirus genome was cloned and maintained as a 230 kb bacterial artificial chromosome (BAC) in E. coli. Transfection of the BAC plasmid into eukaryotic cells led to a productive virus infection. The feasibility to introduce targeted mutations into the BAC cloned virus genome was shown by mutation of the immediate-early 1 gene and generation of a mutant virus. Thus, the complete construction of a mutant herpesvirus genome can now be carried out in a controlled manner prior to the reconstitution of infectious progeny. The described approach should be generally applicable to the mutagenesis of genomes of other large DNA viruses. PMID- 9405687 TI - A system for functional analysis of Ebola virus glycoprotein. AB - Ebola virus causes hemorrhagic fever in humans and nonhuman primates, resulting in mortality rates of up to 90%. Studies of this virus have been hampered by its extraordinary pathogenicity, which requires biosafety level 4 containment. To circumvent this problem, we developed a novel complementation system for functional analysis of Ebola virus glycoproteins. It relies on a recombinant vesicular stomatitis virus (VSV) that contains the green fluorescent protein gene instead of the receptor-binding G protein gene (VSVDeltaG*). Herein we show that Ebola Reston virus glycoprotein (ResGP) is efficiently incorporated into VSV particles. This recombinant VSV with integrated ResGP (VSVDeltaG*-ResGP) infected primate cells more efficiently than any of the other mammalian or avian cells examined, in a manner consistent with the host range tropism of Ebola virus, whereas VSVDeltaG* complemented with VSV G protein (VSVDeltaG*-G) efficiently infected the majority of the cells tested. We also tested the utility of this system for investigating the cellular receptors for Ebola virus. Chemical modification of cells to alter their surface proteins markedly reduced their susceptibility to VSVDeltaG*-ResGP but not to VSVDeltaG*-G. These findings suggest that cell surface glycoproteins with N-linked oligosaccharide chains contribute to the entry of Ebola viruses, presumably acting as a specific receptor and/or cofactor for virus entry. Thus, our VSV system should be useful for investigating the functions of glycoproteins from highly pathogenic viruses or those incapable of being cultured in vitro. PMID- 9405688 TI - N-methyl-D-aspartate receptor activation and visual activity induce elongation factor-2 phosphorylation in amphibian tecta: a role for N-methyl-D-aspartate receptors in controlling protein synthesis. AB - N-methyl-D-aspartate receptor (NMDAR) activation has been implicated in forms of synaptic plasticity involving long-term changes in neuronal structure, function, or protein expression. Transcriptional alterations have been correlated with NMDAR-mediated synaptic plasticity, but the problem of rapidly targeting new proteins to particular synapses is unsolved. One potential solution is synapse specific protein translation, which is suggested by dendritic localization of numerous transcripts and subsynaptic polyribosomes. We report here a mechanism by which NMDAR activation at synapses may control this protein synthetic machinery. In intact tadpole tecta, NMDAR activation leads to phosphorylation of a subset of proteins, one of which we now identify as the eukaryotic translation elongation factor 2 (eEF2). Phosphorylation of eEF2 halts protein synthesis and may prepare cells to translate a new set of mRNAs. We show that NMDAR activation-induced eEF2 phosphorylation is widespread in tadpole tecta. In contrast, in adult tecta, where synaptic plasticity is reduced, this phosphorylation is restricted to short dendritic regions that process binocular information. Biochemical and anatomical evidence shows that this NMDAR activation-induced eEF2 phosphorylation is localized to subsynaptic sites. Moreover, eEF2 phosphorylation is induced by visual stimulation, and NMDAR blockade before stimulation eliminates this effect. Thus, NMDAR activation, which is known to mediate synaptic changes in the developing frog, could produce local postsynaptic alterations in protein synthesis by inducing eEF2 phosphorylation. PMID- 9405689 TI - Targeted deletion of all isoforms of the trkC gene suggests the use of alternate receptors by its ligand neurotrophin-3 in neuronal development and implicates trkC in normal cardiogenesis. AB - We have generated null mutant mice that lack expression of all isoforms encoded by the trkC locus. These mice display a behavioral phenotype characterized by a loss of proprioceptive neurons. Neuronal counts of sensory ganglia in the trkC mutant mice reveal less severe losses than those in NT-3 null mutant mice, strongly suggesting that NT-3, in vivo, may signal through receptors other than trkC. Mice lacking either NT-3 or all trkC receptor isoforms die in the early postnatal period. Histological examination of trkC-deficient mice reveals severe cardiac defects such as atrial and ventricular septal defects, and valvular defects including pulmonic stenosis. Formation of these structures during development is dependent on cardiac neural crest function. The similarities in cardiac defects observed in the trkC and NT-3 null mutant mice indicate that the trkC receptor mediates most NT-3 effects on the cardiac neural crest. PMID- 9405690 TI - Ca2+-dependent and -independent interactions of the isoforms of the alpha1A subunit of brain Ca2+ channels with presynaptic SNARE proteins. AB - Fast neurotransmission requires that docked synaptic vesicles be located near the presynaptic N-type or P/Q-type calcium channels. Specific protein-protein interactions between a synaptic protein interaction (synprint) site on N-type and P/Q-type channels and the presynaptic SNARE proteins syntaxin, SNAP-25, and synaptotagmin are required for efficient, synchronous neurotransmitter release. Interaction of the synprint site of N-type calcium channels with syntaxin and SNAP-25 has a biphasic calcium dependence with maximal binding at 10-20 microM. We report here that the synprint sites of the BI and rbA isoforms of the alpha1A subunit of P/Q-type Ca2+ channels have different patterns of interactions with synaptic proteins. The BI isoform of alpha1A specifically interacts with syntaxin, SNAP-25, and synaptotagmin independent of Ca2+ concentration and binds with high affinity to the C2B domain of synaptotagmin but not the C2A domain. The rbA isoform of alpha1A interacts specifically with synaptotagmin and SNAP-25 but not with syntaxin. Binding of synaptotagmin to the rbA isoform of alpha1A is Ca2+ dependent, with maximum affinity at 10-20 microM Ca2+. Although the rbA isoform of alpha1A binds well to both the C2A and C2B domains of synaptotagmin, only the interaction with the C2A domain is Ca2+-dependent. These differential, Ca2+ dependent interactions of Ca2+ channel synprint sites with SNARE proteins may modulate the efficiency of transmitter release triggered by Ca2+ influx through these channels. PMID- 9405691 TI - A new mechanism of sound generation in songbirds. AB - Our current understanding of the sound-generating mechanism in the songbird vocal organ, the syrinx, is based on indirect evidence and theoretical treatments. The classical avian model of sound production postulates that the medial tympaniform membranes (MTM) are the principal sound generators. We tested the role of the MTM in sound generation and studied the songbird syrinx more directly by filming it endoscopically. After we surgically incapacitated the MTM as a vibratory source, zebra finches and cardinals were not only able to vocalize, but sang nearly normal song. This result shows clearly that the MTM are not the principal sound source. The endoscopic images of the intact songbird syrinx during spontaneous and brain stimulation-induced vocalizations illustrate the dynamics of syringeal reconfiguration before phonation and suggest a different model for sound production. Phonation is initiated by rostrad movement and stretching of the syrinx. At the same time, the syrinx is closed through movement of two soft tissue masses, the medial and lateral labia, into the bronchial lumen. Sound production always is accompanied by vibratory motions of both labia, indicating that these vibrations may be the sound source. However, because of the low temporal resolution of the imaging system, the frequency and phase of labial vibrations could not be assessed in relation to that of the generated sound. Nevertheless, in contrast to the previous model, these observations show that both labia contribute to aperture control and strongly suggest that they play an important role as principal sound generators. PMID- 9405692 TI - Role of left inferior prefrontal cortex in retrieval of semantic knowledge: a reevaluation. AB - A number of neuroimaging findings have been interpreted as evidence that the left inferior frontal gyrus (IFG) subserves retrieval of semantic knowledge. We provide a fundamentally different interpretation, that it is not retrieval of semantic knowledge per se that is associated with left IFG activity but rather selection of information among competing alternatives from semantic memory. Selection demands were varied across three semantic tasks in a single group of subjects. Functional magnetic resonance imaging signal in overlapping regions of left IFG was dependent on selection demands in all three tasks. In addition, the degree of semantic processing was varied independently of selection demands in one of the tasks. The absence of left IFG activity for this comparison counters the argument that the effects of selection can be attributed solely to variations in degree of semantic retrieval. Our findings suggest that it is selection, not retrieval, of semantic knowledge that drives activity in the left IFG. PMID- 9405693 TI - A sensory brain map for each behavior? AB - Multiple brain maps are commonly found in virtually every vertebrate sensory system. Although their functional significance is generally relatively little understood, they seem to specialize in processing distinct sensory parameters. Nevertheless, to yield the stimulus features that ultimately elicit the adaptive behavior, it appears that information streams have to be combined across maps. Results from current lesion experiments in the electrosensory system, however, suggest an alternative possibility. Inactivations of different maps of the first order electrosensory nucleus in electric fish, the electrosensory lateral line lobe, resulted in markedly different behavioral deficits. The centromedial map is both necessary and sufficient for a particular electrolocation behavior, the jamming avoidance response, whereas it does not affect the communicative response to external electric signals. Conversely, the lateral map does not affect the jamming avoidance response but is necessary and sufficient to evoke communication behavior. Because the premotor pathways controlling the two behaviors in these fish appear to be separated as well, this system illustrates that sensory-motor control of different behaviors can occur in strictly segregated channels from the sensory input of the brain all through to its motor output. This might reflect an early evolutionary stage where multiplication of brain maps can satisfy the demand on processing a wider range of sensory signals ensuing from an enlarged behavioral repertoire, and bridging across maps is not yet required. PMID- 9405694 TI - Neurotrophin-3 signals redistribute RNA in neurons. AB - The translocation of specific mRNAs to dendrites and their potential for locally regulated translation are likely to serve as an effector in neuronal plasticity. Whether translation in dendrites is regulated by delivery of the RNA to sites of plasticity or a stationary pool of localized RNA undergoes enhanced translational efficiency is not clear. We show that RNA can translocate into dendrites in response to NT-3. RNA granules were visualized in cultured rat cortical neurons using the dye SYTO 14, which labels poly-ribosome complexes. Long before the morphological effects of NT-3 appeared, there was increased distal translocation of labeled complexes. This effect was blocked by K252a, a potent inhibitor of tyrosine kinase receptors. Therefore, neurons can utilize extracellular signals to alter the distribution of protein synthetic machinery via the active transport of RNA granules. PMID- 9405695 TI - In vitro-generated neural precursors participate in mammalian brain development. AB - During embryogenesis, pluripotent stem cells segregate into daughter lineages of progressively restricted developmental potential. In vitro, this process has been mimicked by the controlled differentiation of embryonic stem cells into neural precursors. To explore the developmental potential of these cell-culture-derived precursors in vivo, we have implanted them into the ventricles of embryonic rats. The transplanted cells formed intraventricular neuroepithelial structures and migrated in large numbers into the brain tissue. Embryonic-stem-cell-derived neurons, astrocytes, and oligodendrocytes incorporated into telencephalic, diencephalic, and mesencephalic regions and assumed phenotypes indistinguishable from neighboring host cells. These observations indicate that entirely in vitro generated neural precursors are able to respond to environmental signals guiding cell migration and differentiation and have the potential to reconstitute neuronal and glial lineages in the central nervous system. PMID- 9405696 TI - Interactive cloning with the SH3 domain of N-src identifies a new brain specific ion channel protein, with homology to eag and cyclic nucleotide-gated channels. AB - We have isolated a novel cDNA, that appears to represent a new class of ion channels, by using the yeast two-hybrid system and the SH3 domain of the neural form of Src (N-src) as a bait. The encoded polypeptide, BCNG-1, is distantly related to cyclic nucleotide-gated channels and the voltage-gated channels, Eag and H-erg. BCNG-1 is expressed exclusively in the brain, as a glycosylated protein of approximately 132 kDa. Immunohistochemical analysis indicates that BCNG-1 is preferentially expressed in specific subsets of neurons in the neocortex, hippocampus, and cerebellum, in particular pyramidal neurons and basket cells. Within individual neurons, the BCNG-1 protein is localized to either the dendrites or the axon terminals depending on the cell type. Southern blot analysis shows that several other BCNG-related sequences are present in the mouse genome, indicating the emergence of an entire subfamily of ion channel coding genes. These findings suggest the existence of a new type of ion channel, which is potentially able to modulate membrane excitability in the brain and could respond to regulation by cyclic nucleotides. PMID- 9405697 TI - Glutamate transporter currents in bergmann glial cells follow the time course of extrasynaptic glutamate. AB - Glutamate transporters in the central nervous system are expressed in both neurons and glia, they mediate high affinity, electrogenic uptake of glutamate, and they are associated with an anion conductance that is stoichiometrically uncoupled from glutamate flux. Although a complete cycle of transport may require 50-100 ms, previous studies suggest that transporters can alter synaptic currents on a much faster time scale. We find that application of L-glutamate to outside out patches from cerebellar Bergmann glia activates anion-potentiated glutamate transporter currents that activate in <1 ms, suggesting an efficient mechanism for the capture of extrasynaptic glutamate. Stimulation in the granule cell layer in cerebellar slices elicits all or none alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionate receptor and glutamate transporter currents in Bergmann glia that have a rapid onset, suggesting that glutamate released from climbing fiber terminals escapes synaptic clefts and reaches glial membranes shortly after release. Comparison of the concentration dependence of both alpha-amino-3-hydroxy 5-methyl-4-isoxazolepropionate receptor and glutamate transporter kinetics in patches with the time course of climbing fiber-evoked responses indicates that the glutamate transient at Bergmann glial membranes reaches a lower concentration than attained in the synaptic cleft and remains elevated in the extrasynaptic space for many milliseconds. PMID- 9405698 TI - Vascular imprints of neuronal activity: relationships between the dynamics of cortical blood flow, oxygenation, and volume changes following sensory stimulation. AB - Modern functional neuroimaging methods, such as positron-emission tomography (PET), optical imaging of intrinsic signals, and functional MRI (fMRI) utilize activity-dependent hemodynamic changes to obtain indirect maps of the evoked electrical activity in the brain. Whereas PET and flow-sensitive MRI map cerebral blood flow (CBF) changes, optical imaging and blood oxygenation level-dependent MRI map areas with changes in the concentration of deoxygenated hemoglobin (HbR). However, the relationship between CBF and HbR during functional activation has never been tested experimentally. Therefore, we investigated this relationship by using imaging spectroscopy and laser-Doppler flowmetry techniques, simultaneously, in the visual cortex of anesthetized cats during sensory stimulation. We found that the earliest microcirculatory change was indeed an increase in HbR, whereas the CBF increase lagged by more than a second after the increase in HbR. The increased HbR was accompanied by a simultaneous increase in total hemoglobin concentration (Hbt), presumably reflecting an early blood volume increase. We found that the CBF changes lagged after Hbt changes by 1 to 2 sec throughout the response. These results support the notion of active neurovascular regulation of blood volume in the capillary bed and the existence of a delayed, passive process of capillary filling. PMID- 9405700 TI - A genetic screen for mutations that disrupt an auditory response in Drosophila melanogaster. AB - Hearing is one of the last sensory modalities to be subjected to genetic analysis in Drosophila melanogaster. We describe a behavioral assay for auditory function involving courtship among groups of males triggered by the pulse component of the courtship song. In a mutagenesis screen for mutations that disrupt the auditory response, we have recovered 15 mutations that either reduce or abolish this response. Mutant audiograms indicate that seven mutants reduced the amplitude of the response at all intensities. Another seven abolished the response altogether. The other mutant, 5L3, responded only at high sound intensities, indicating that the threshold was shifted in this mutant. Six mutants were characterized in greater detail. 5L3 had a general courtship defect; courtship of females by 5L3 males also was affected strongly. 5P1 males courted females normally but had reduced success at copulation. 5P1 and 5N18 showed a significant decrement in olfactory response, indicating that the defects in these mutations are not specific to the auditory pathway. Two other mutants, 5M8 and 5N30, produced amotile sperm although in 5N30 this phenotype was genetically separable from the auditory phenotype. Finally, a new adult circling behavior phenotype, the pirouette phenotype, associated with massive neurodegeneration in the brain, was discovered in two mutants, 5G10 and 5N18. This study provides the basis for a genetic and molecular dissection of auditory mechanosensation and auditory behavior. PMID- 9405699 TI - Postnatal mouse subventricular zone neuronal precursors can migrate and differentiate within multiple levels of the developing neuraxis. AB - The mammalian subventricular zone (SVZ) of the lateral wall of the forebrain ventricle retains a population of proliferating neuronal precursors throughout life. Neuronal precursors born in the postnatal and adult SVZ migrate to the olfactory bulb where they differentiate into interneurons. Here we tested the potential of mouse postnatal SVZ precursors in the environment of the embryonic brain: (i) a ubiquitous genetic marker, (ii) a neuron-specific transgene, and (iii) a lipophilic-dye were used to follow the fate of postnatal day 5-10 SVZ cells grafted into embryonic mouse brain ventricles at day 15 of gestation. Graft derived cells were found at multiple levels of the neuraxis, including septum, thalamus, hypothalamus, and in large numbers in the midbrain inferior colliculus. We observed no integration into the cortex. Neuronal differentiation of graft derived cells was demonstrated by double-staining with neuron-specific beta tubulin antibodies, expression of the neuron-specific transgene, and the dendritic arbors revealed by the lipophilic dye. We conclude that postnatal SVZ cells can migrate through and differentiate into neurons within multiple embryonic brain regions other than the olfactory bulb. PMID- 9405701 TI - Very short-term plasticity in hippocampal synapses. AB - Hippocampal pyramidal neurons often fire in bursts of action potentials with short interspike intervals (2-10 msec). These high-frequency bursts may play a critical role in the functional behavior of hippocampal neurons, but synaptic plasticity at such short times has not been carefully studied. To study synaptic modulation at very short time intervals, we applied pairs of stimuli with interpulse intervals ranging from 7 to 50 msec to CA1 synapses isolated by the method of minimal stimulation in hippocampal slices. We have identified three components of short-term paired-pulse modulation, including (i) a form of synaptic depression manifested after a prior exocytotic event, (ii) a form of synaptic depression that does not depend on a prior exocytotic event and that we postulate is based on inactivation of presynaptic N-type Ca2+ channels, and (iii) a dependence of paired-pulse facilitation on the exocytotic history of the synapse. PMID- 9405702 TI - Targeted gene deletion of heme oxygenase 2 reveals neural role for carbon monoxide. AB - Neuronal nitric oxide synthase (nNOS) generates NO in neurons, and heme-oxygenase 2 (HO-2) synthesizes carbon monoxide (CO). We have evaluated the roles of NO and CO in intestinal neurotransmission using mice with targeted deletions of nNOS or HO-2. Immunohistochemical analysis demonstrated colocalization of nNOS and HO-2 in myenteric ganglia. Nonadrenergic noncholinergic relaxation and cyclic guanosine 3',5' monophosphate elevations evoked by electrical field stimulation were diminished markedly in both nNOSDelta/Delta and HO-2(Delta)/Delta mice. In wild-type mice, NOS inhibitors and HO inhibitors partially inhibited nonadrenergic noncholinergic relaxation. In nNOSDelta/Delta animals, NOS inhibitors selectively lost their efficacy, and HO inhibitors were inactive in HO 2(Delta)/Delta animals. PMID- 9405703 TI - Orphanin FQ acts as an anxiolytic to attenuate behavioral responses to stress. AB - Orphanin FQ (OFQ, Nociceptin) is a recently discovered 17-amino acid neuropeptide that is structurally related to the opioid peptides but does not bind opioid receptors. OFQ has been proposed to act as an anti-opioid peptide, but its widespread sites of action in the brain suggest that it may have more general functions. Here we show that OFQ plays an important role in higher brain functions because it can act as an anxiolytic to attenuate the behavioral inhibition of animals acutely exposed to stressful/anxiogenic environmental conditions. OFQ anxiolytic-like effects were consistent across several behavioral paradigms generating different types of anxiety states in animals (light-dark preference, elevated plus-maze, exploratory behavior of an unfamiliar environment, pharmacological anxiogenesis, operant conflict) and were observed at low nonsedating doses (0.1-3 nmol, intracerebroventricular). Like conventional anxiolytics, OFQ interfered with regular sensorimotor function at high doses (>3 nmol). Our results show that an important role of OFQ is to act as an endogenous regulator of acute anxiety responses. OFQ, probably in concert with other major neuropeptides, exerts a modulatory role on the central integration of stressful stimuli and, thereby, may modulate anxiety states generated by acute stress. PMID- 9405705 TI - Neurosteroids: deficient cognitive performance in aged rats depends on low pregnenolone sulfate levels in the hippocampus. AB - Pregnenolone sulfate (PREG S) is synthesized in the nervous system and is a major neurosteroid in the rat brain. Its concentrations were measured in the hippocampus and other brain areas of single adult and aged (22-24 month-old) male Sprague-Dawley rats. Significantly lower levels were found in aged rats, although the values were widely scattered and reached, in about half the animals, the same range as those of young ones. The spatial memory performances of aged rats were investigated in two different spatial memory tasks, the Morris water maze and Y maze. Performances in both tests were significantly correlated and, accompanied by appropriate controls, likely evaluated genuine memory function. Importantly, individual hippocampal PREG S and distance to reach the platform in the water maze were linked by a significant correlation, i.e., those rats with lower memory deficit had the highest PREG S levels, whereas no relationship was found with the PREG S content in other brain areas (amygdala, prefrontal cortex, parietal cortex, striatum). Moreover, the memory deficit of cognitively impaired aged rats was transiently corrected after either intraperitoneal or bilateral intrahippocampal injection of PREG S. PREG S is both a gamma-aminobutyric acid antagonist and a positive allosteric modulator at the N-methyl-D-aspartate receptor, and may reinforce neurotransmitter system(s) that decline with age. Indeed, intracerebroventricular injection of PREG S was shown to stimulate acetylcholine release in the adult rat hippocampus. In conclusion, it is proposed that the hippocampal content of PREG S plays a physiological role in preserving and/or enhancing cognitive abilities in old animals, possibly via an interaction with central cholinergic systems. Thus, neurosteroids should be further studied in the context of prevention and/or treatment of age-related memory disorders. PMID- 9405704 TI - The phosphoprotein DARPP-32 mediates cAMP-dependent potentiation of striatal N methyl-D-aspartate responses. AB - The signal transduction pathway underlying the cAMP-dependent modulation of rat striatal N-methyl-D-aspartate (NMDA) responses was investigated by using the two electrode voltage-clamp technique. In oocytes injected with rat striatal poly(A)+ mRNA, activation of cAMP-dependent protein kinase (PKA) by forskolin potentiated NMDA responses. Inhibition of protein phosphatase 1 (PP1) and/or protein phosphatase 2A (PP2A) by the specific inhibitor calyculin A occluded the PKA mediated potentiation of striatal NMDA responses, suggesting that the PKA effect was mediated by inhibition of a protein phosphatase. Coinjection of oocytes with striatal mRNA and antisense oligodeoxynucleotides directed against the protein phosphatase inhibitor DARPP-32 dramatically reduced the PKA enhancement of NMDA responses. NMDA responses recorded from oocytes injected with rat hippocampal poly(A)+ mRNA were not affected by stimulation of PKA. When oocytes were coinjected with rat hippocampal poly(A)+ mRNA plus complementary RNA coding for DARPP-32, NMDA responses were potentiated after stimulation of PKA. The results provide evidence that DARPP-32, which is enriched in the striatum, may participate in the signaling between the two major afferent striatal pathways, the glutamatergic and the dopaminergic projections, by the cAMP-dependent regulation of striatal NMDA currents. PMID- 9405706 TI - Block of transmitter release by botulinum C1 action on syntaxin at the squid giant synapse. AB - Electrophysiological, morphological, and biochemical approaches were combined to study the effect of the presynaptic injection of the light chain of botulinum toxin C1 into the squid giant synapse. Presynaptic injection was accompanied by synaptic block that occurred progressively as the toxin filled the presynaptic terminal. Neither the presynaptic action potential nor the Ca2+ currents in the presynaptic terminal were affected by the toxin. Biochemical analysis of syntaxin moiety in squid indicates that the light chain of botulinum toxin C1 lyses syntaxin in vitro, suggesting that this was the mechanism responsible for synaptic block. Ultrastructure of the injected synapses demonstrates an enormous increase in the number of presynaptic vesicles, suggesting that the release rather than the docking of vesicles is affected by biochemical lysing of the syntaxin molecule. PMID- 9405707 TI - Transient "deafness" accompanies auditory development during metamorphosis from tadpole to frog. AB - During metamorphosis, ranid frogs shift from a purely aquatic to a partly terrestrial lifestyle. The central auditory system undergoes functional and neuroanatomical reorganization in parallel with the development of new sound conduction pathways adapted for the detection of airborne sounds. Neural responses to sounds can be recorded from the auditory midbrain of tadpoles shortly after hatching, with higher rates of synchronous neural activity and lower sharpness of tuning than observed in postmetamorphic animals. Shortly before the onset of metamorphic climax, there is a brief "deaf" period during which no auditory activity can be evoked from the midbrain, and a loss of connectivity is observed between medullary and midbrain auditory nuclei. During the final stages of metamorphic development, auditory function and neural connectivity are restored. The acoustic communication system of the adult frog emerges from these periods of anatomical and physiological plasticity during metamorphosis. PMID- 9405708 TI - Predominance of the alpha1D subunit in L-type voltage-gated Ca2+ channels of hair cells in the chicken's cochlea. AB - The voltage-gated Ca2+ channels that effect tonic release of neurotransmitter from hair cells have unusual pharmacological properties: unlike most presynaptic Ca2+ channels, they are sensitive to dihydropyridines and therefore are L-type. To characterize these Ca2+ channels, we investigated the expression of L-type alpha1 subunits in hair cells of the chicken's cochlea. In PCRs with five different pairs of degenerate primers, we always obtained alpha1D products, but only once an alpha1C product and never an alpha1S product. A full-length alpha1D mRNA sequence was assembled from overlapping PCR products; the predicted amino acid sequence of the alpha1D subunit was about 90% identical to those of the mammalian alpha1D subunits. In situ hybridization confirmed that the alpha1D mRNA is present in hair cells. By using a quantitative PCR assay, we determined that the alpha1D mRNA is 100-500 times more abundant than the alpha1C mRNA. We conclude that most, if not all, voltage-gated Ca2+ channels in hair cells contain an alpha1D subunit. Furthermore, we propose that the alpha1D subunit plays a hitherto undocumented role at tonic synapses. PMID- 9405709 TI - Hair cell-specific splicing of mRNA for the alpha1D subunit of voltage-gated Ca2+ channels in the chicken's cochlea. AB - The L-type voltage-gated Ca2+ channels that control tonic release of neurotransmitter from hair cells exhibit unusual electrophysiological properties: a low activation threshold, rapid activation and deactivation, and a lack of Ca2+ dependent inactivation. We have inquired whether these characteristics result from cell-specific splicing of the mRNA for the L-type alpha1D subunit that predominates in hair cells of the chicken's cochlea. The alpha1D subunit in hair cells contains three uncommon exons: one encoding a 26-aa insert in the cytoplasmic loop between repeats I and II, an alternative exon for transmembrane segment IIIS2, and a heretofore undescribed exon specifying a 10-aa insert in the cytoplasmic loop between segments IVS2 and IVS3. We propose that the alternative splicing of the alpha1D mRNA contributes to the unusual behavior of the hair cell's voltage-gated Ca2+ channels. PMID- 9405710 TI - Apparent mtDNA heteroplasmy in Alzheimer's disease patients and in normals due to PCR amplification of nucleus-embedded mtDNA pseudogenes. AB - In an unprecedented finding, Davis et al. [Davis, R. E., Miller, S., Herrnstadt, C., Ghosh, S. S., Fahy, E., Shinobu, L. A., Galasko, D., Thal, L. J., Beal, M. F., Howell, N. & Parker, W. D., Jr. (1997) Proc. Natl. Acad. Sci. USA 94, 4526 4531] used an unusual DNA isolation method to show that healthy adults harbor a specific population of mutated mitochondrial cytochrome c oxidase (COX) genes that coexist with normal mtDNAs. They reported that this heteroplasmic population was present at a level of 10-15% in the blood of normal individuals and at a significantly higher level (20-30%) in patients with sporadic Alzheimer's disease. We provide compelling evidence that the DNA isolation method employed resulted in the coamplification of authentic mtDNA-encoded COX genes together with highly similar COX-like sequences embedded in nuclear DNA ("mtDNA pseudogenes"). We conclude that the observed heteroplasmy is an artifact. PMID- 9405712 TI - Construction of a high-affinity receptor site for dihydropyridine agonists and antagonists by single amino acid substitutions in a non-L-type Ca2+ channel. AB - The activity of L-type Ca2+ channels is increased by dihydropyridine (DHP) agonists and inhibited by DHP antagonists, which are widely used in the therapy of cardiovascular disease. These drugs bind to the pore-forming alpha1 subunits of L-type Ca2+ channels. To define the minimal requirements for DHP binding and action, we constructed a high-affinity DHP receptor site by substituting a total of nine amino acid residues from DHP-sensitive L-type alpha1 subunits into the S5 and S6 transmembrane segments of domain III and the S6 transmembrane segment of domain IV of the DHP-insensitive P/Q-type alpha1A subunit. The resulting chimeric alpha1A/DHPS subunit bound DHP antagonists with high affinity in radioligand binding assays and was inhibited by DHP antagonists with high affinity in voltage clamp experiments. Substitution of these nine amino acid residues yielded 86% of the binding energy of the L-type alpha1C subunit and 92% of the binding energy of the L-type alpha1S subunit for the high-affinity DHP antagonist PN200-110. The activity of chimeric Ca2+ channels containing alpha1A/DHPS was increased 3.5 +/- 0.7-fold by the DHP agonist (-)Bay K8644. The effect of this agonist was stereoselective as in L-type Ca2+ channels since (+) Bay K8644 inhibited the activity of alpha1A/DHPS. The results show conclusively that DHP agonists and antagonists bind to a single receptor site at which they have opposite effects on Ca2+ channel activity. This site contains essential components from both domains III and IV, consistent with a domain interface model for binding and allosteric modulation of Ca2+ channel activity by DHPs. PMID- 9405711 TI - Ancient mtDNA sequences in the human nuclear genome: a potential source of errors in identifying pathogenic mutations. AB - Nuclear-localized mtDNA pseudogenes might explain a recent report describing a heteroplasmic mtDNA molecule containing five linked missense mutations dispersed over the contiguous mtDNA CO1 and CO2 genes in Alzheimer's disease (AD) patients. To test this hypothesis, we have used the PCR primers utilized in the original report to amplify CO1 and CO2 sequences from two independent rho degrees (mtDNA less) cell lines. CO1 and CO2 sequences amplified from both of the rho degrees cells, demonstrating that these sequences are also present in the human nuclear DNA. The nuclear pseudogene CO1 and CO2 sequences were then tested for each of the five "AD" missense mutations by restriction endonuclease site variant assays. All five mutations were found in the nuclear CO1 and CO2 PCR products from rho degrees cells, but none were found in the PCR products obtained from cells with normal mtDNA. Moreover, when the overlapping nuclear CO1 and CO2 PCR products were cloned and sequenced, all five missense mutations were found, as well as a linked synonymous mutation. Unlike the findings in the original report, an additional 32 base substitutions were found, including two in adjacent tRNAs and a two base pair deletion in the CO2 gene. Phylogenetic analysis of the nuclear CO1 and CO2 sequences revealed that they diverged from modern human mtDNAs early in hominid evolution about 770,000 years before present. These data would be consistent with the interpretation that the missense mutations proposed to cause AD may be the product of ancient mtDNA variants preserved as nuclear pseudogenes. PMID- 9405713 TI - Loperamide: a positive modulator for store-operated calcium channels? AB - The depletion of inositol trisphosphate-sensitive intracellular pools of calcium causes activation of store-operated calcium (SOC) channels. Loperamide at 10-30 microM has no effect on intracellular calcium levels alone, but augments calcium levels in cultured cells when SOC channels have been activated. In HL-60 leukemic cells, the apparent positive modulatory effect of loperamide on SOC channels occurs when these channels have been activated after ATP, thapsigargin, or ionomycin-elicited depletion of calcium from intracellular storage sites. Loperamide has no effect when levels of intracellular calcium are elevated through a mechanism not involving SOC channels by using sphingosine. Loperamide caused augmentation of intracellular calcium levels after activation of SOC channels in NIH 3T3 fibroblasts, astrocytoma 1321N cells, smooth muscle DDT-MF2 cells, RBL-2H3 mast cells, and pituitary GH4C1 cells. Only in astrocytoma cells did loperamide cause an elevation in intracellular calcium in the absence of activation of SOC channels. The augmentation of intracellular calcium elicited by loperamide in cultured cells was dependent on extracellular calcium and was somewhat resistant to agents (SKF 96365, miconazole, clotrimazole, nitrendipine, and trifluoperazine) that in the absence of loperamide effectively blocked SOC channels. It appears that loperamide augments influx of calcium through activated SOC channels. PMID- 9405714 TI - Inactivation of calcium-activated chloride channels in smooth muscle by calcium/calmodulin-dependent protein kinase. AB - To determine the mechanisms responsible for the termination of Ca2+-activated Cl- currents (ICl(Ca)), simultaneous measurements of whole cell currents and intracellular Ca2+ concentration ([Ca2+]i) were made in equine tracheal myocytes. In nondialyzed cells, or cells dialyzed with 1 mM ATP, ICl(Ca) decayed before the [Ca2+]i decline, whereas the calcium-activated potassium current decayed at the same rate as [Ca2+]i. Substitution of AMP-PNP or ADP for ATP markedly prolonged the decay of ICl(Ca), resulting in a rate of current decay similar to that of the fall in [Ca2+]i. In the presence of ATP, dialysis of the calmodulin antagonist W7, the Ca2+/calmodulin-dependent kinase II (CaMKII) inhibitor KN93, or a CaMKII specific peptide inhibitor the rate of ICl(Ca) decay was slowed and matched the [Ca2+]i decline, whereas H7, a nonspecific kinase inhibitor with low affinity for CaMKII, was without effect. When a sustained increase in [Ca2+]i was produced in ATP dialyzed cells, the current decayed completely, whereas in cells loaded with 5'-adenylylimidodiphosphate (AMP-PNP), KN93, or the CaMKII inhibitory peptide, ICl(Ca) did not decay. Slowly decaying currents were repeatedly evoked in ADP- or AMP-PNP-loaded cells, but dialysis of adenosine 5'-O-(3-thiotriphosphate) or okadaic acid resulted in a smaller initial ICl(Ca), and little or no current (despite a normal [Ca2+]i transient) with a second stimulation. These data indicate that CaMKII phosphorylation results in the inactivation of calcium activated chloride channels, and that transition from the inactivated state to the closed state requires protein dephosphorylation. PMID- 9405716 TI - Slow and fast dietary proteins differently modulate postprandial protein accretion. AB - The speed of absorption of dietary amino acids by the gut varies according to the type of ingested dietary protein. This could affect postprandial protein synthesis, breakdown, and deposition. To test this hypothesis, two intrinsically 13C-leucine-labeled milk proteins, casein (CAS) and whey protein (WP), of different physicochemical properties were ingested as one single meal by healthy adults. Postprandial whole body leucine kinetics were assessed by using a dual tracer methodology. WP induced a dramatic but short increase of plasma amino acids. CAS induced a prolonged plateau of moderate hyperaminoacidemia, probably because of a slow gastric emptying. Whole body protein breakdown was inhibited by 34% after CAS ingestion but not after WP ingestion. Postprandial protein synthesis was stimulated by 68% with the WP meal and to a lesser extent (+31%) with the CAS meal. Postprandial whole body leucine oxidation over 7 h was lower with CAS (272 +/- 91 micromol.kg-1) than with WP (373 +/- 56 micromol.kg-1). Leucine intake was identical in both meals (380 micromol.kg-1). Therefore, net leucine balance over the 7 h after the meal was more positive with CAS than with WP (P < 0.05, WP vs. CAS). In conclusion, the speed of protein digestion and amino acid absorption from the gut has a major effect on whole body protein anabolism after one single meal. By analogy with carbohydrate metabolism, slow and fast proteins modulate the postprandial metabolic response, a concept to be applied to wasting situations. PMID- 9405715 TI - 11beta-hydroxysteroid dehydrogenase type 1 knockout mice show attenuated glucocorticoid-inducible responses and resist hyperglycemia on obesity or stress. AB - Glucocorticoid hormones, acting via nuclear receptors, regulate many metabolic processes, including hepatic gluconeogenesis. It recently has been recognized that intracellular glucocorticoid concentrations are determined not only by plasma hormone levels, but also by intracellular 11beta-hydroxysteroid dehydrogenases (11beta-HSDs), which interconvert active corticosterone (cortisol in humans) and inert 11-dehydrocorticosterone (cortisone in humans). 11beta-HSD type 2, a dehydrogenase, thus excludes glucocorticoids from otherwise nonselective mineralocorticoid receptors in the kidney. Recent data suggest the type 1 isozyme (11beta-HSD-1) may function as an 11beta-reductase, regenerating active glucocorticoids from circulating inert 11-keto forms in specific tissues, notably the liver. To examine the importance of this enzyme isoform in vivo, mice were produced with targeted disruption of the 11beta-HSD-1 gene. These mice were unable to convert inert 11-dehydrocorticosterone to corticosterone in vivo. Despite compensatory adrenal hyperplasia and increased adrenal secretion of corticosterone, on starvation homozygous mutants had attenuated activation of the key hepatic gluconeogenic enzymes glucose-6-phosphatase and phosphoenolpyruvate carboxykinase, presumably, because of relative intrahepatic glucocorticoid deficiency. The 11beta-HSD-1 -/- mice were found to resist hyperglycamia provoked by obesity or stress. Attenuation of hepatic 11beta-HSD-1 may provide a novel approach to the regulation of gluconeogenesis. PMID- 9405717 TI - Antisense oligonucleotides against rat brain alpha1E DNA and its atrial homologue decrease T-type calcium current in atrial myocytes. AB - Low voltage-activated, or T-type, calcium currents are important regulators of neuronal and muscle excitability, secretion, and possibly cell growth and differentiation. The gene (or genes) coding for the pore-forming subunit of low voltage-activated channel proteins has not been unequivocally identified. We have used reverse transcription-PCR to identify partial clones from rat atrial myocytes that share high homology with a member of the E class of calcium channel genes. Antisense oligonucleotides targeting one of these partial clones (raE1) specifically block the increase in T-current density that normally results when atrial myocytes are treated with insulin-like growth factor 1 (IGF-1). Antisense oligonucleotides targeting portions of the neuronal rat alpha1E sequence, which are not part of the clones detected in atrial tissue, also block the IGF-1 induced increase in T-current, suggesting that the high homology to alpha1E seen in the partial clone may be present in the complete atrial sequence. The basal T current expressed in these cells is also blocked by antisense oligonucleotides, which is consistent with the notion that IGF-1 up-regulates the same gene that encodes the basal current. These results support the hypothesis that a member of the E class of calcium channel genes encodes a low voltage-activated calcium channel in atrial myocytes. PMID- 9405718 TI - Anchoring of protein kinase A facilitates hormone-mediated insulin secretion. AB - Impaired insulin secretion is a characteristic of non-insulin-dependent diabetes mellitus (NIDDM). One possible therapeutic agent for NIDDM is the insulinotropic hormone glucagon-like peptide 1 (GLP-1). GLP-1 stimulates insulin secretion through several mechanisms including activation of protein kinase A (PKA). We now demonstrate that the subcellular targeting of PKA through association with A kinase-anchoring proteins (AKAPs) facilitates GLP-1-mediated insulin secretion. Disruption of PKA anchoring by the introduction of anchoring inhibitor peptides or expression of soluble AKAP fragments blocks GLP-1 action in primary islets and cAMP-responsive insulin secretion in clonal beta cells (RINm5F). Displacement of PKA also prevented cAMP-mediated elevation of intracellular calcium suggesting that localized PKA phosphorylation events augment calcium flux. PMID- 9405719 TI - Leaf-specifically expressed genes for polypeptides destined for chloroplasts with domains of sigma70 factors of bacterial RNA polymerases in Arabidopsis thaliana. AB - Genes for sigma-like factors of bacterial-type RNA polymerase have not been characterized from any multicellular eukaryotes, although they probably play a crucial role in the expression of plastid photosynthesis genes. We have cloned three distinct cDNAs, designated SIG1, SIG2, and SIG3, for polypeptides possessing amino acid sequences for domains conserved in sigma70 factors of bacterial RNA polymerases from the higher plant Arabidopsis thaliana. Each gene is present as one copy per haploid genome without any additional sequences hybridized in the genome. Transient expression assays using green fluorescent protein demonstrated that N-terminal regions of the SIG2 and SIG3 ORFs could function as transit peptides for import into chloroplasts. Transcripts for all three SIG genes were detected in leaves but not in roots, and were induced in leaves of dark-adapted plants in rapid response to light illumination. Together with results of our previous analysis of tissue-specific regulation of transcription of plastid photosynthesis genes, these results indicate that expressed levels of the genes may influence transcription by regulating RNA polymerase activity in a green tissue-specific manner. PMID- 9405720 TI - Differentially regulated NADPH:cytochrome P450 oxidoreductases in parsley. AB - Two NADPH:cytochrome P450 oxidoreductases (CPRs) from parsley (Petroselinum crispum) were cloned, and the complete proteins were expressed and functionally identified in yeast. The two enzymes, designated CPR1 and CPR2, are 80% identical in amino acid sequence with one another and about 75% identical with CPRs from several other plant species. The mRNA accumulation patterns for CPR1 and CPR2 in fungal elicitor-treated or UV-irradiated cultured parsley cells and in developing or infected parsley plants were compared with those for cinnamate 4-hydroxylase (C4H), one of the most abundant CPR-dependent P450 enzymes in plants. All treatments strongly induced the mRNAs for C4H and CPR1 but not for CPR2, suggesting distinct metabolic roles of CPR1 and CPR2 and a functional relationship between CPR1 and C4H. PMID- 9405721 TI - An Arabidopsis mutant that requires increased calcium for potassium nutrition and salt tolerance. AB - Potassium (K+) nutrition and salt tolerance are key factors controlling plant productivity. However, the mechanisms by which plants regulate K+ nutrition and salt tolerance are poorly understood. We report here the identification of an Arabidopsis thaliana mutant, sos3 (salt-overly-sensitive 3), which is hypersensitive to Na+ and Li+ stresses. The mutation is recessive and is in a nuclear gene that maps to chromosome V. The sos3 mutation also renders the plant unable to grow on low K+. Surprisingly, increased extracellular Ca2+ suppresses the growth defect of sos3 plants on low K+ or 50 mM NaCl. In contrast, high concentrations of external Ca2+ do not rescue the growth of the salt hypersensitive sos1 mutant on low K+ or 50 mM NaCl. Under NaCl stress, sos3 seedlings accumulated more Na+ and less K+ than the wild type. Increased external Ca2+ improved K+/Na+ selectivity of both sos3 and wild-type plants. However, this Ca2+ effect in sos3 is more than twice as much as that in the wild type. In addition to defining the first plant mutant with an altered calcium response, these results demonstrate that the SOS3 locus is essential for K+ nutrition, K+/Na+ selectivity, and salt tolerance in higher plants. PMID- 9405722 TI - Brain wave recognition of words. AB - Electrical and magnetic brain waves of seven subjects under three experimental conditions were recorded for the purpose of recognizing which one of seven words was processed. The analysis consisted of averaging over trials to create prototypes and test samples, to both of which Fourier transforms were applied, followed by filtering and an inverse transformation to the time domain. The filters used were optimal predictive filters, selected for each subject and condition. Recognition rates, based on a least-squares criterion, varied widely, but all but one of 24 were significantly different from chance. The two best were above 90%. These results show that brain waves carry substantial information about the word being processed under experimental conditions of conscious awareness. PMID- 9405723 TI - Preliminary description of the cranium of Proteopithecus sylviae, an Egyptian late Eocene anthropoidean primate. AB - Recent discovery of crania, dentitions, and postcrania of a primitive anthropoidean primate, Proteopithecus sylviae, at the late Eocene L-4l quarry in the Fayum, Egypt, provides evidence of a new taxonomic family of early African higher primates, the Proteopithecidae. This family could be part of the basal radiation that produced the New World platyrrhine primates, or it could be unrelated to any subsequent lineages. Although no larger than a small callitrichid or a dwarf lemur, this tiny primate already possessed many of the derived features of later anthropoids and was a diurnal and probably dimorphic species. In dental formula and other dental proportions, as well as in known postcranial features, Proteopithecus more nearly resembles platyrrhines than does any other Old World higher primate. The small size of the Proteopithecus cranium demonstrates that the defining cranial characteristics of Anthropoidea did not arise as a consequence of an increase in size during derivation from earlier prosimians. PMID- 9405724 TI - Perspectives: some roles of mechanical usage, muscle strength, and the mechanostat in skeletal physiology, disease, and research. PMID- 9405725 TI - Calcium supplementation suppresses bone resorption in early postmenopausal women. AB - In order to establish whether calcium supplementation suppresses bone resorption in early postmenopausal women and whether any response is related to calcium absorption status, we studied 22 healthy women (median age 52 years) all within 5 years of the menopause. Urine was collected between 9.00 p.m. and 9.00 a.m., and 9.00 a.m. and 9.00 p.m., (2 days) and a fasting blood and spot urine sample was obtained at 9 a.m. On the first day, 5 microCi of 45Ca in 250 ml water with 20 mg calcium carrier as the chloride was given at 9.00 a.m. and a further blood sample was obtained at 10.00 a.m. to measure calcium absorption. A 1 g calcium load was given at 9.00 p.m., immediately before the second 24-hour urine collection. There was a rise in plasma ionized calcium (1.18 +/- 0.010 mmol/liter versus 1. 21 +/- 0.011 mmol/liter, P < 0.01) and a fall in plasma PTH (4.2 +/- 0.34 pmol/liter versus 3.5 +/- 0.31 pmol/liter, P < 0.01) from baseline after the calcium load, and a trend for the magnitude of the change in PTH to be inversely related to calcium absorption (r = -0.33, P = 0.13). In the fasting spot urine samples, there were falls in hydroxyproline (OHPr/Cr; 14.6 +/- 0.71 versus 12.6 +/- 0.83, P < 0.001), pyridinoline (Pyr/Cr; 75 +/- 2.8 versus 70 +/- 3.5, P < 0.05), and deoxypyridinoline (Dpd/Cr; 22.7 +/- 1.2 versus 19.5 +/- 1. 1, P < 0.005) after the calcium load. The calcium load suppressed urinary Dpd/Cr between 9.00 p.m. and 9.00 a.m. (P < 0.005), but not between 9.00 a.m. and 9.00 p.m. We conclude that acute administration of a 1 g calcium load suppresses bone resorption in early postmenopausal women, probably by decreasing PTH secretion. PMID- 9405727 TI - Differing lumbar vertebral mineralization rates in ambulatory pediatric patients with osteogenesis imperfecta. AB - The purpose of this study was to evaluate serial changes in bone mineral density (BMD) of the lumbar spine in individual children and adolescents with untreated osteogenesis imperfecta (OI) using dual X-ray absorptiometry (DXA). Twenty-seven pediatric patients with OI who had no historical or radiographic evidence of lumbar fracture, required no assistive device for mobility, and were taking no medications known to affect skeletal mineralization during the study period comprised the investigational group. Absolute BMD and age- and gender-matched BMD (Z-scores) were assessed relative to standard parameters of growth (height, weight, age, height adjusted for age and gender and body surface area) and severity of disease (lifetime fracture rate). The spinal mineralization rate (SMR) between examinations for 15 patients with more than one measurement (n = 20 intervals) was expressed as the magnitude of the change in BMD Z-score per year. Both BMD and BMD Z-score were closely correlated with height, height Z-score, weight and body surface area and were inversely related to fracture rate (P < 0.001 for all comparisons). BMD was also highly correlated with patient age (P < 0.001). Stepwise regression analysis showed that together height Z-score and lifetime fracture rate improved the prediction of BMD Z-score (r = 0. 71; P < 0.001). SMRs ranged from -0.5 to 3.5. The average change in SMR between sequential measurements was 168% for the five children who had more than two DXA examinations. Linear regression showed a significant negative correlation between SMR and height Z-score (r = -0.79, P < 0.001). We conclude that vertebral body size is a critical determinant of BMD and BMD Z-score in OI because DXA results are expressed per unit area, not per unit volume. Pediatric patients with OI mineralize their lumbar vertebrae at rates similar to healthy children but tend to lag behind in overall mineralization. The rate of mineralization at any age appears to be related to the patient's height (adjusted for age- and gender matched controls) and inversely related to the patient's lifetime rate of fractures. Our data suggest that vertebral mineralization in children with OI is related primarily to rapid increases in vertebral volume and only secondarily to increases in vertebral mineral density. PMID- 9405726 TI - Cytokine release from marrow mononuclear cells is negatively correlated to cortical elasticity in non-osteoporotic postmenopausal women. AB - Activation of bone remodeling is likely to be under the control of mechanical factors acting, in part, through soluble local factors. We therefore investigated a relationship between cytokine production by marrow cells and bone elasticity. We studied 36 non-osteoporotic postmenopausal women undergoing hip arthroplasty for hip arthrosis (mean age: 68 +/- 8 years; lumbar BMD Z-score: +0.54 +/- 0.33 SD). Adherent marrow mononuclear cells were cultured for 48 hours with autologous plasma, and supernatants were harvested for PGE2, IL-1, TNF-alpha, and IL-6 measurements. Femoral neck cortical bones were removed during surgery for cortical histomorphometric evaluation and determination of elasticity indices (C33) using ultrasonic transmission method. In this nonosteoporotic population, femoral neck longitudinal elasticity indices were inversely correlated to both cortical thickness (r = -0.58, P < 0.01) and cortical porosity (r = -0.33, P < 0.01). The longitudinal elasticity indices were also negatively correlated to basal IL-1 and TNF-alpha release by adherent mononuclear marrow cells (r = -0.59, P < 0.01; r = -0.60, P < 0.01, respectively). However, no relationship was found between the three cytokines tested and either cortical thickness or porosity. These data show a link between cortical biomechanical properties and local factors involved in bone remodeling. We suggest that increased bone elasticity decreases transmission of strain, which in turn decreases cytokine release from marrow cells. However, whether cytokines influence bone elasticity or vice versa remains to be demonstrated. PMID- 9405728 TI - Biochemical markers as predictors of bone mineral density changes after GnRH agonist treatment. AB - To evaluate bone biochemical markers as predictors of the efficacy of a hormone replacement therapy (HRT), we studied the bone changes induced by the cessation and return of ovarian function in 28 patients treated for 6 months with a GnRH agonist. This model reproduced the effects observed in postmenopausal women with high bone turnover treated with HRT. At the end of the treatment, Z scores were 1.8 +/- 0.3 for Crosslaps (CTx) and deoxypyridinoline (D-Pyr), and 1.1 +/- 0.2 for bone alkaline phosphatase (B-ALP) and osteocalcin (OC). This indicated an imbalance in bone remodeling with a high bone resorption. Bone mineral density (BMD) fell by 4.2 +/- 2.5%. The changes in BMD between the 6th and 12th months were 0. 34 +/- 2.24 and -1.73 +/- 3.25% at the lumbar spine and the femoral neck, respectively. Biochemical markers except urinary calcium and hydroxyproline measured at 6 months were positively correlated with the BMD changes at the lumbar spine. After the resumption of menstruation, 13 of 28 women displayed positive spine BMD changes between the 6th and 12th months; in this group, bone biochemical markers measured at 6 months were significantly higher (P = 0.02). Stepwise regression analysis showed that the association of B-ALP and D-Pyr measured at 6 months explained 40% of BMD variance and the association of B-ALP, PTH, and estradiol 56%. We conclude that measuring individual biochemical bone markers can help to predict the bone effect of an increase in the circulating estradiol in women with ovarian deficiency. PMID- 9405729 TI - Effects of postmenopausal hormone replacement therapy with and without vitamin D3 on circulating levels of 25-hydroxyvitamin D and 1,25-dihydroxyvitamin D. AB - The effects of postmenopausal hormone replacement therapy (HRT) and vitamin D3 on vitamin D metabolites (25OHD and 1,25(OH)2D) were studied in a population-based prospective 1-year study. The serum concentrations of intact parathyroid hormone (PTH), calcium, and phosphate were also studied. A total of 72 women were randomized into four treatment groups: HRT group (sequential combination of 2 mg estradiol valerate and 1 mg cyproterone acetate), Vit D3 group (vitamin D3 300 IU/day + calcium lactate 500 mg/day), HRT + Vit D3 group (both above) and placebo group (calcium lactate 500 mg/day). Serum samples were taken in March-April, when vitamin D formation from sunlight in Finland is minimal after the dark winter. Serum concentrations of 25OHD increased in the Vit D3 group (33.5%, P < 0. 001) and in the HRT + Vit D3 group (38.2%, P < 0.001) but had not changed significantly in the HRT and placebo groups at the 1-year follow-up examination. Serum concentrations of calcitriol (1, 25(OH)2D) increased, however, only in the HRT group (23.7%, P < 0. 05), and remained unchanged in other groups. Serum concentrations of PTH decreased by 23.2% (P < 0.05) in the placebo group, but did not change significantly in the other three groups. The concentrations of serum calcium increased in the nonhormone groups (P < 0.001), whereas serum phosphate concentrations decreased in the hormone groups (P < 0.05 and 0.001). Our results confirm the positive effect of 1 year of HRT on serum calcitriol. Vitamin D3 supplementation increased 25OHD concentrations, but did not affect calcitriol concentrations even though the initial levels were low. Interestingly, the combination of HRT and vitamin D3 did not increase serum calcitriol concentrations as much as HRT alone. PMID- 9405730 TI - Bone mineral density in children and adolescents with diabetes mellitus type 1 of recent onset. AB - There is still controversy over the impact of diabetes control and duration on bone mass and growth parameters in children and adolescents with insulin dependent diabetes mellitus (IDDM). The aim of this study was to assess bone mineral density (BMD) at axial and appendicular sites, in children with noncomplicated IDDM of recent onset, and its relation to metabolic control and auxological parameters (weight, height, and puberal stage). Fifty-five young Spanish IDDM, otherwise healthy patients (26 males, aged (SD 9.7 +/- 4.3 years) and 29 females, aged (SD 11.2 +/- 3.8 years) were studied. Duration of diabetes was 1-13.8 years. Two hundred eighty-two age-matched, healthy, Spanish children served as controls. HbA1 was assayed by high pressure liquid chromatography (HPLC) and BMD was measured using dual X-ray absorptiometry (DXA) densitometry at the spine and forearm. Results showed a Gaussian BMD distribution of patients according to sex and age, without sexual-stage differences. There was no correlation between BMD and glycated hemoglobin (average life disease or last HbA1 values) or duration of the disease; moreover, no differences in bone mass were found between <3 and >/=3 years of disease duration. Diabetes impact index (mean HbA1 x duration of disease in months) showed no significant influence of diabetes control on BMD. We could not demonstrate any impact of diabetes on BMD and growth parameters in children with IDDM of short duration. PMID- 9405731 TI - Urinary excretion of type I collagen degradation products in healthy women and osteoporotic patients with vertebral and hip fractures. AB - We have evaluated both the effect of normal aging and menopause on urinary CrossLapstrade mark (u-CTx) excretion and the bone resorption status by u-CTx in patients with vertebral fracture and hip fracture. In 246 healthy women, 76 patients with vertebral fracture, and 63 patients with hip fracture, u-CTx excretion was measured by ELISA. The age-related changes of u-CTx in healthy females reflected the marked increase of bone resorption associated with modeling at childhood. The values in the subgroups of postmenopausal women 1-3 years since menopause and 8% and to reduce cardiovascular risk factors. The medical charge data suggest that investment in clinical systems to improve diabetes care may benefit both payers and patients. PMID- 9405906 TI - Visceral fat and cardiovascular risk factors in Chinese NIDDM patients. AB - OBJECTIVE: The interrelations between obesity, glucose intolerance, hypertension, dyslipidemia, and insulin resistance are well recognized. These relationships are of particular interest in Hong Kong's Chinese population, in whom increasing affluence has coincided with a marked increase in the prevalence of NIDDM. We designed a pilot study to examine the relationships between visceral fat and cardiovascular risk factors in Chinese NIDDM patients. RESEARCH DESIGN AND METHODS: We studied 21 Chinese NIDDM patients whose visceral fat was quantified by magnetic resonance imaging. Cardiovascular risk factors including plasma lipids and 24-h ambulatory blood pressure (BP) were measured. In addition, insulin resistance was determined by a short insulin tolerance test (SITT). RESULTS: Increased visceral adiposity was significantly correlated with plasma triglycerides (r = 0.63, P = 0.004), the total cholesterol/HDL cholesterol ratio (r = 0.61, P = 0.008), the urinary albumin/creatinine ratio (r = 0.49, P = 0.04), and decreased insulin sensitivity as measured by the SITT (r = 0.47, P = 0.03). When the data were analyzed by tertiles, increasing visceral fat area was associated with higher plasma triglycerides, lower HDL cholesterol, and a smaller plasma glucose decrement during the SITT. In addition, the diurnal rhythm in BP and heart rate tended to be best preserved in those with the least visceral obesity. CONCLUSIONS: This pilot study demonstrates that visceral fat accumulation is associated with dyslipidemia, hypertension, insulin resistance, and albuminuria in Chinese patients with NIDDM. PMID- 9405907 TI - Comparison of diabetes diagnostic categories in the U.S. population according to the 1997 American Diabetes Association and 1980-1985 World Health Organization diagnostic criteria. AB - OBJECTIVE: To compare the 1997 American Diabetes Association (ADA) and the 1980 1985 World Health Organization (WHO) diagnostic criteria in categorization of the diabetes diagnostic status of adults in the U.S. RESEARCH DESIGN AND METHODS: Analyses are based on a probability sample of the U.S. population age 40-74 years in the 1988-1994 Third National Health and Nutrition Examination Survey (NHANES III). People with diabetes diagnosed before the survey were identified by questionnaire. For 2,844 people without diagnosed diabetes, fasting plasma glucose was obtained after an overnight 9 to < 24-h fast, HbA1c was measured, and a 2-h oral glucose tolerance test was administered. RESULTS: Prevalence of diagnosed diabetes in this age-group is 7.9%. Prevalence of undiagnosed diabetes is 4.4% by ADA criteria and 6.4% by WHO criteria. The net change of -2.0% occurs because 1.0% are classified as having undiagnosed diabetes by ADA criteria but have impaired or normal glucose tolerance by WHO criteria, and 3.0% are classified as having impaired fasting glucose or normal fasting glucose by ADA criteria but have undiagnosed diabetes by WHO criteria. Prevalence of impaired fasting glucose is 10.1% (ADA), compared with 15.6% for impaired glucose tolerance (WHO). For those with undiagnosed diabetes by ADA criteria, 62.1% are above the normal range for HbA1c compared with 47.1% by WHO criteria. Mean HbA1c is 7.07% for undiagnosed diabetes by ADA criteria and 6.58% by WHO criteria. CONCLUSIONS: The number of people with undiagnosed diabetes by ADA criteria is lower than that by WHO criteria. However, those individuals classified by ADA criteria are more hyperglycemic, with higher HbA1c values and a greater proportion of values above the normal range. This fact, together with the simplicity of obtaining a fasting plasma glucose value, may result in the detection of a greater proportion of people with undiagnosed diabetes in clinical practice using the new ADA diagnostic criteria. PMID- 9405908 TI - Results of a placebo-controlled study of the metabolic effects of the addition of metformin to sulfonylurea-treated patients. Evidence for a central role of adipose tissue. AB - OBJECTIVE: To define the metabolic effects of metformin in the treatment of NIDDM and to evaluate potential mechanisms for its ability to improve glycemic control. RESEARCH DESIGN AND METHODS: Sulfonylurea-treated patients, with inadequate glycemic control, were treated with metformin in either a placebo-controlled or open fashion. Measurements were made of 1) fasting and postprandial plasma glucose, insulin, and free fatty acid (FFA) concentrations; 2) glucose appearance and disappearance rates measured overnight with 3-[3H]glucose; and 3) plasma FFA concentrations during a 45-min infusion period at relatively low (approximately 60 pmol/l) insulin concentrations. RESULTS: Mean +/- SE hourly plasma glucose, insulin, and FFA concentrations were similar before and after treatment in the placebo group. In contrast, mean hourly plasma glucose concentrations were significantly lower (P < 0.005) after metformin treatment in both the placebo controlled and open-label groups (-3.9 +/- 1.0 and -4.4 +/- 0.8 mmol/l, respectively). Similarly, day-long hourly FFA levels were lower (P < 0.005) following metformin in the placebo-controlled and open-label groups (-87 +/- 35 and -136 +/- 31 mumol/l, respectively). Plasma insulin concentrations did not change with treatment in any group. Overnight glucose turnover studies indicated that neither the rate of glucose appearance (hepatic glucose production) or glucose disappearance changed significantly with treatment in the placebo or metformin groups. Because plasma glucose concentration was much lower after metformin treatment, overnight glucose metabolic clearance rate was significantly (P < 0.001) lower in this group. Finally, plasma FFA concentrations in response to a low-dosage insulin infusion (5 mU.m-2.min-1) were significantly lower after metformin as compared with the placebo-treated group (P < 0.001). CONCLUSIONS: Metformin treatment was associated with significantly lower day-long plasma glucose and FFA concentrations. Although overnight hepatic glucose production was unchanged following treatment with metformin, the overnight glucose metabolic clearance rate significantly increased. Given these findings, it is suggested that at least part of the antihyperglycemic effect of metformin is due to an increase in glucose uptake, secondary to a decrease in release of FFA from adipose tissue, and lower circulating FFA concentrations. PMID- 9405909 TI - Nighttime insulin kinetics and glycemic control in type 1 diabetes patients following administration of an intermediate-acting lispro preparation. AB - OBJECTIVE: To determine insulin kinetics and overnight glycemic control after bedtime administration of a new intermediate-acting insulin preparation called neutral protamine lispro (NPL). RESEARCH DESIGN AND METHODS: We studied 12 patients with well-controlled type 1 diabetes. The study had a double-blind, randomized, crossover design. After a lead-in period of 10-14 days two experiments were carried out with an interval of 2-7 days. During these experiments overnight insulin kinetics and fasting blood glucose levels were studied after evening administration of NPH insulin and NPL. Blood glucose levels < 3.8 mmol/l were treated by means of a variable infusion of a 20% glucose solution. RESULTS: A trend toward a shorter time to peak insulin concentration was observed after administration of NPL (P = 0.07). No differences between NPH and NPL were detected in the total area under the curve (AUC) for insulin, in insulin levels before breakfast, or in glucose levels before breakfast (P = 0.5, 0.6, and 0.4, respectively). CONCLUSIONS: We detected no major differences between NPH and NPL in the total AUC for insulin, prebreakfast glucose levels, or prebreakfast insulin levels. Therefore, we conclude that NPH and NPL are equally effective in controlling overnight glycemia. PMID- 9405910 TI - GLP-1 tablet in type 2 diabetes in fasting and postprandial conditions. AB - OBJECTIVE: To examine the absorption of glucagon-like peptide (GLP)-1(7-36) amide from the buccal mucosa of type 2 diabetic patients. Previously, the effects of the peptide have been studied following intravenous and subcutaneous injection. Now, a mucoadhesive, biodegradable buccal GLP-1 tablet (9 mm) containing 119 nmol has been developed as a possible alternative to injection. RESEARCH DESIGN AND METHODS: A total of 10 type 2 diabetic patients received a single tablet under fasting conditions and before a standard meal in this randomized placebo controlled study. RESULTS: The mean peak GLP-1 concentration was 125.1 pmol/l and occurred 30 min after application. The mean placebo-adjusted area under the curve was 5,334 min pmol/l, consistent with a relative bioavailability of 6% vs. intravenous injection and 42% vs. subcutaneous injection. The half-life of total peptide activity after buccal administration was 17 min. The placebo-adjusted glucose concentrations decreased by 1.4 mmol/l in fasting experiments and by 4.2 mmol/l after a standard mixed meal. In the fasting state at 30 min, plasma insulin increased by 185% and glucagon decreased by 20%, consistent with the increase in plasma GLP-1 concentrations. The peptide exerted a significant insulinotropic effect during meals (calculated as an insulinogenic index, 0-120 min; 84.1 vs. 45.7 in placebo experiments). CONCLUSIONS: Potentially therapeutic plasma levels of GLP-1 were achieved after administration of a single buccal tablet in type 2 diabetic patients. The peptide had a marked glucose-lowering effect during the first 2 h. This new GLP-1 tablet may become a feasible alternative treatment for type 2 diabetic patients, although a more prolonged pharmacokinetic profile is required. PMID- 9405912 TI - Effects of Trp64Arg mutation in the beta 3-adrenergic receptor gene on weight loss, body fat distribution, glycemic control, and insulin resistance in obese type 2 diabetic patients. AB - OBJECTIVE: To investigate the effects of Trp64Arg mutation in the beta 3 adrenergic receptor gene on weight loss, body fat distribution, glycemic control, and insulin resistance in obese type 2 diabetic patients. RESEARCH DESIGN AND METHODS: We measured body weight, waist-to-hip ratio (WHR), adjusted resting metabolic rate, fasting blood glucose, fasting serum insulin levels, insulin resistance index (fasting glucose x fasting insulin/22.5), and HbA1c levels before and after 12 weeks of obesity treatment in 61 obese women with type 2 diabetes. The MvaI polymorphism of the beta 3-adrenergic receptor gene was determined by polymerase chain reaction-restriction fragment length polymorphism analysis. RESULTS: Of obese type 2 diabetic patients, those with the mutation (n = 24) had a higher WHR (P < 0.001), a lower adjusted metabolic rate, and higher blood glucose levels, serum insulin levels, insulin resistance index (P < 0.001), and HbA1c levels (P = 0.016). Furthermore, patients with the mutation had smaller decreases in body weight, WHR, insulin resistance index, and HbA1c levels after the weight-loss program compared with patients without the mutation (n = 37), even though food intake, exercise, and serum thyroid hormone levels were similar in both groups. CONCLUSIONS: These present findings show that the Trp64Arg allele of the beta 3-adrenergic receptor gene may predict difficulty in losing body weight, lowering WHR, and improving glycemic control and insulin resistance in obese patients with type 2 diabetes. PMID- 9405911 TI - Hyperhomocyst(e)inemia and endothelial dysfunction in IDDM. AB - OBJECTIVE: While elevated blood levels of homocyst(e)ine represent an independent risk factor for macrovascular disease, we assessed the link between hyperhomocyst(e)inemia and diabetic microvascular diseases. RESEARCH DESIGN AND METHODS: Plasma levels of homocyst(e)ine and thrombomodulin (TM), markers of endothelial cell damage, were measured before and 3 h after oral methionine loading in 75 patients with IDDM and 40 healthy control subjects matched for sex and age. Exclusion criteria were hyperlipidemia, hypertension, smoking, or positive family history for cardiovascular disease. RESULTS: IDDM patients had higher pre- and postload plasma levels of homocyst(e)ine than did healthy control subjects (12.0 vs. 7.7 mumol/l and 27.6 vs. 16.0 mumol/l; P < 0.001). Of 75 IDDM patients, 26 had plasma homocyst(e)ine levels above the normal range (means +/- 2 SD of values obtained in the control group). These IDDM patients with hyperhomocyst(e)inemia had higher plasma TM levels (62.2 vs. 38.2 ng/ml, P < 0.001), higher albumin excretion rates (485 vs. 115 mg/l, P < 0.005), and a higher prevalence of late diabetic complications (nephropathy, 76 vs. 33%; retinopathy, 69 vs. 51%; neuropathy, 57 vs. 41%; and macroangiopathy, 57 vs. 33%) compared with IDDM patients with normal plasma homocyst(e)ine. In vitro experiments with human umbilical vein cells showed an increased release of TM into the culture supernatant only when endothelial cells were pretreated with advanced glycation end product (AGE)-albumin before L-homocystine was added. A synergistic action of homocyst(e)ine and AGEs might contribute to vascular complications in patients with diabetes. CONCLUSIONS: Hyperhomocyst(e)inemia is common in nephropathic diabetic patients and may contribute to the enhanced morbidity and mortality from cardiovascular diseases characteristically observed in IDDM patients with diabetic nephropathy. PMID- 9405913 TI - Reduction of albumin excretion rate in normotensive microalbuminuric type 2 diabetic patients during long-term simvastatin treatment. AB - OBJECTIVE: To study the long-term effects of simvastatin on urinary albumin excretion rate (AER) in normotensive microalbuminuric type 2 diabetic patients with hypercholesterolemia. RESEARCH DESIGN AND METHODS: A total of 19 normotensive microalbuminuric hypercholesterolemic type 2 diabetic patients entered a double-blind crossover study for 2 years, receiving either simvastatin (20 mg/day) or placebo (each treatment for 1 year). RESULTS: Simvastatin significantly decreased plasma cholesterol (total and LDL) after 52 weeks of treatment. A concomitant significant decrease of AER (25% from basal) with no significant changes in creatinine clearance was observed during the same period. CONCLUSIONS: Our data are in keeping with the hypothesis that simvastatin might be used as an additional means to preserve renal function in microalbuminuric hypercholesterolemic type 2 diabetic patients. PMID- 9405914 TI - Serum autoantibodies against sulfatide and phospholipid in NIDDM patients with diabetic neuropathy. AB - OBJECTIVE: We investigated the presence of antisulfatide and antiphospholipid antibodies and the relationship between these antibodies and the results of quantitative tests of nerve function in NIDDM patients with diabetic neuropathy. RESEARCH DESIGN AND METHODS: Antisulfatide and antiphospholipid antibodies were measured in serum samples obtained from 68 NIDDM patients with diabetic neuropathy by an enzyme-linked immunosorbent assay (ELISA). Each patient was classified into one of three groups based on the combined neuropathy score (determined by the symptom score, the results of autonomic nerve function tests, and the vibration perception test), as follows: mild (n = 26), moderate (n = 22), and severe (n = 20). Nerve conduction studies were performed in a subgroup of 37 patients. RESULTS: The antisulfatide antibody was detected in 1 (4%) of 26 patients in the mild group, 4 (18%) of 22 patients in the moderate group, and 8 (40%) of 20 patients in the severe group (P < 0.01 vs. mild group). The antiphospholipid antibody was detected in none of the patients in the mild group, 8 (36%) of 22 patients in the moderate group (P < 0.001 vs. mild group), and 6 (30%) of 20 patients in the severe group (P < 0.01 vs. mild group). The threshold amplitude, determined by the vibration perception test, was significantly higher in antibody-positive patients than in antibody-negative patients: antisulfatide antibody, 55.9 +/- 46.8 microns (n = 13) vs. 22.9 +/- 13.7 microns (n = 55), P < 0.001; antiphospholipid antibody, 47.2 +/- 32.5 microns (n = 14) vs. 24.5 +/- 23.2 microns (n = 54), P < 0.01. The conduction velocity of the sural nerve was slower in the antisulfatide antibody-positive group (37.9 +/- 11.1 m/s, n = 12) than in the antisulfatide antibody-negative group (45.2 +/- 6.0 m/s, n = 19) (P < 0.05). CONCLUSIONS: These results suggest that autoimmune nerve destruction may be involved in diabetic neuropathy in NIDDM patients. PMID- 9405915 TI - Antibodies to GAD and glycemic control in recent-onset IDDM. AB - OBJECTIVE: To analyze the effect of antibodies to glutamic acid decarboxylase (GAD65Ab) and islet cells (ICA512Ab) on glycemic control early in IDDM. RESEARCH DESIGN AND METHODS: GAD65Ab and ICA512Ab were measured twice in 35 patients (10 male, 25 female; age 10-40 years) initially within 2 years of diagnosis and again 1 year later. The glycosylated hemoglobin was measured one to four times each year, and the average glycosylated hemoglobin for the preceding year was calculated each time the antibodies were measured. RESULTS: The mean HbA1 at the time of the initial evaluation was 8.04 +/- 0.30 (reference range 4.7-7.3% for nondiabetic patients), the average GAD65Ab index was 0.735 +/- 0.306, and the mean ICA512Ab index was 1.94 +/- 0.65. The GAD65Ab index correlated with HbA1 (r = 0.41, P < 0.025), whereas the ICA512Ab index did not (r = 0.13). One year later, the mean GAD65Ab index was 0.94 +/- 0.34, the mean ICA512Ab index was 1.04 +/- 0.40, and the mean HbA1 was 9.03 +/- 0.30. The GAD65Ab index correlated with HbA1 (r = 0.61 P < 0.001), whereas the ICA512Ab index did not (r = -0.06). Stratification of patients into tertiles according to the average GAD65 index revealed, at the follow-up evaluation, that the better glycemic control in the lowest GAD65Ab tertile was accomplished with significantly less insulin (0.43 +/- 0.08 U/kg for the lowest tertile vs. 0.71 +/- 0.09 and 0.64 +/- 0.09 for the middle and highest tertiles, respectively; P < 0.05). CONCLUSIONS: In summary, patients with IDDM and low GAD65Ab have better glycemic control even though they require less insulin. The ICA512Ab index, however, fails to correlate with glycemia. PMID- 9405916 TI - Gait abnormalities in diabetic neuropathy. AB - OBJECTIVE: To investigate the effect of peripheral neuropathy on gait in diabetic patients. RESEARCH DESIGN AND METHODS: Gait analysis was performed in the following groups matched for age, sex, and BMI: 20 normal healthy control subjects (NC), 20 non-neuropathic diabetic control subjects (DC), 20 neuropathic diabetic subjects (DN), and 20 neuropathic diabetic subjects with a history of foot ulceration (DNU). All subjects with orthopedic foot problems were excluded from the study. The following gait parameters were investigated: 1) walking speed; 2) stance phase duration; 3) joint angles and moment arms for the ankle, knee, and hip joints in both sagittal and frontal planes; 4) the components of the ground reaction force (GRF) vector; and 5) the ankle, knee, and hip joint moments originating from the GRF vector in both planes. RESULTS: There were no statistical differences in any of the parameters studied between the NC and DC groups. Walking speed was significantly slower in the DNU group compared with the two control groups (P < 0.02). The maximum knee joint angle was smaller in the sagittal plane for the DNU group compared with the DC group values (P < 0.05). The maximum value of the vertical component of GRF was found to be higher (P < 0.03) in the two control groups compared with the DNU group. The maximum value of the anteroposterior forces was also found to be higher (P < 0.001) in the DC group compared with the DNU group. The maximum frontal plane ankle joint moment was significantly higher (P < 0.05) in the DN compared with the NC group. CONCLUSIONS: Diabetic subjects with peripheral neuropathy demonstrate alterations in some gait parameters during walking. These alterations could facilitate foot injuries, thus contributing to frequent foot ulceration. PMID- 9405917 TI - Diabetes mellitus and exercise. American Diabetes Association. PMID- 9405918 TI - American Diabetes Association Annual Meeting, 1997. Obesity, diabetes prevention, and type 1 diabetes. PMID- 9405919 TI - alpha-Lipoic acid in NIDDM patients with cardiac autonomic neuropathy. PMID- 9405920 TI - Plasma endothelin in normal and diabetic pregnancy. PMID- 9405921 TI - High lipoprotein(a) levels in type 1 and type 2 diabetic patients with macroalbuminuria. PMID- 9405922 TI - Noncardiogenic pulmonary edema in a patient with diabetic ketoacidosis. PMID- 9405923 TI - Intimal-medical thickness of the carotid artery in NIDDM patients. PMID- 9405924 TI - Hyper-insulin response in a patient with depression. Changes in insulin resistance during recovery from depression. PMID- 9405925 TI - Fasting in Ramadan and the diabetic patient. PMID- 9405926 TI - Apolipoprotein A-IV and E polymorphisms in children with IDDM. PMID- 9405927 TI - The usefulness of three biallelic restriction fragment length polymorphisms versus a polymorphic dinucleotide tandem repeat polymorphism at the low-density lipoprotein receptor gene locus for diagnosis of familial hypercholesterolemia. AB - Indirect molecular diagnosis of familial hypercholesterolemia is possible based on genetic linkage analysis of DNA polymorphisms at the low-density-lipoprotein receptor gene locus in family studies. The use of biallelic restriction fragment length polymorphisms, however, is restricted by their low degree of heterogeneity, and therefore several of these markers have to be combined. To overcome these restrictions we examined the value and applicability of an (AT)n tandem repeat polymorphism at the 3' untranslated region of the gene, alone and in combination with three biallelic restriction fragment length polymorphisms in 35 independent healthy subjects and in familial hypercholesterolemia families with 23 parents and 52 children. For each family one of the parents had the clinical diagnosis of familial hypercholesterolemia. The probands were genotyped using the TaqI, HincII and NcoI restriction fragment length polymorphism and the (AT)n tandem repeat polymorphism of the gene. The heterozygosity index (0.60) and the polymorphism information content (PIC value) (0.53) of the ATn repeat polymorphism were higher compared to each single biallelic restriction fragment length polymorphism (heterozygosity index 0.26-0.54; PIC value 0.24-0.36). The combined PIC value of all three biallelic restriction fragment length polymorphisms (0.79) was comparable to the combination of the HincII and the ATn polymorphism (0.74). Using these two markers, a definitive molecular diagnosis could be made in 36 children from 15 parents compared to just 12 parents and their children using the three biallelic restriction fragment length polymorphisms. We conclude that the ATn tandem repeat polymorphisms is useful for indirect molecular diagnosis of familial hypercholesterolemia in affected families. PMID- 9405928 TI - Characterization of markers flanking the human SP-B locus. AB - We have previously identified a SP-B length polymorphism that appears with higher frequency in the RDS population (Biochem. J., 305, 1995, p583). This polymorphism encompasses a fairly large region, thus it is difficult to distinguish between variants with small size differences. Because of the importance of SP-B in normal lung function and the association of this SP-B polymorphism with RDS, we wished to identify and characterize polymorphic markers linked to the SP-B locus that would allow better resolution of SP-B alleles. In this report we a) characterized a novel (AAGG)n linked SP-B microsatellite marker; b) determined linkage of published markers with the SP-B locus and also determined the distance of each marker from the SP-B locus using medium and high resolution radiation hybrid panels; c) determined heterozygosity index and PIC values of the novel and known markers in various populations; and d) determined haplotypes using CEPH families. The availability of these SP-B linked markers/haplotypes will facilitate population and family based association studies. We are hopeful that the information gained will help to unravel the genetic complexity of RDS and respiratory diseases with regards to the SP-B locus. PMID- 9405929 TI - Diabetic complications and the genetics of signal transduction. A study of retinopathy in NIDDM. AB - Cytosolic low molecular weight acid phosphatase (ACP1) is a high polymorphic phosphotyrosine-protein-phosphatase involved in signal transduction. In NIDDM subjects we have found that ACP1 genotype is a highly significant predictor of retinopathy, suggesting that genetic variability of signal transduction may have an important role in the susceptibility to this complication. Adenosine deaminase, ABO blood groups and several clinical variables have been also considered. The results point out the importance of interactions between genetic systems. Among non-genetic variables dislipidemia and treatment with insulin are significantly associated with retinopathy. PMID- 9405930 TI - Prevalence of hepatitis C infection and schistosomiasis in Egyptian patients with hepatocellular carcinoma. AB - Hepatitis C virus (HCV) infection prevalence was determined in 34 hepatocellular carcinoma (HCC) patients with underlying liver cirrhosis. Serum alfa-fetoprotein level was found to be more than 1000 ng/ml (3153 +/- 1451) in those patients. Anti-HCV antibodies were detected in 84% of patients sera negative for hepatitis B surface antigen (HBsAg), while HCV RNA was only detected in 28% of the sera of those patients. Schistosomiasis antibodies were found in 92% of the anti-HCV positive HBsAg negative sera of HCC patients, while they were only detected in 61% of sera of control non HCC patients with anti-HCV antibodies and without HBsAg. HCV and hepatitis B virus (HBV) coinfection was found in 16% of the studied group, based on antibodies in sera, and in 9% based on the presence of HCV RNA. HBV (HBsAg) was found, on its own, in sera of 5.9% of the patients studied. These results show that HCV and schistosomiasis play a major role in the development of hepatocellular carcinoma in Egypt. PMID- 9405931 TI - Serum LDH and ALP isozyme activities in mice bearing solid Ehrlich carcinoma and/or treated with the maximum tolerated dose (MTD) of aloin. AB - Determination of total LDH and ALP activities and their isozyme patterns in the sera of normal and tumour-bearing animals treated with aloin, a natural anthraquinone with potential antitumour activity, was carried out at 3, 6 and 9 weeks of treatment. Treatment of normal mice with the MTD of aloin (50 mg/Kg b.w.) showed non-significant changes in serum total LDH and ALP activities along with their isozymes throughout the experimental periods. In untreated tumour bearing animals, serum LDH activity and its isozymes: LDH1-LDH5 showed highly significant increases (192, 32.4, 25.2, 24.7, 29.2 and 30.6%, respectively) after 3 weeks. Highly significant inhibition was recorded in serum total ALP activity and its intestinal and bone isozymes (64, 100 and 56%, respectively), while liver ALP isozyme was increased by 82.3%. Treatment of tumour-bearing mice with the MTD of aloin manifested a significant gradual improvement in both enzyme activities and their isozymes, which were normalized at the end of the experiment (9 weeks), with the exception of intestinal ALP isozyme. All results were reported in comparison to the normal control group. PMID- 9405932 TI - Serum cytokines and tumour markers in patients with non-small cell carcinoma of the lung. AB - We compared the serum levels of several cytokines with established tumour markers in 24 patients with non-small cell lung cancer (NSCLC) and 31 patients with benign lung disease (BLD). Cytokine levels were measured using in-house double determinant sandwich ELISAs and tumour markers by a variety of established techniques. There was no correlation between serum cytokines and expression of cytokeratins 18 and 19, MUCI and carcinoembryonic antigen. While no significant difference was observed in any of the cytokines between patients with NSCLC and BLD, patients with metastatic tumour had a significantly higher level of serum tumour necrosis factor alpha and interleukin 10 than those with localised disease (P < 0.015 and P < 0.05 respectively). The serum levels of granulocyte macrophage colony stimulating factor and interferon gamma were no different between these groups. These results suggest immunological effects of NSCLC which tends towards impaired cell mediated immunity in patients with metastatic disease. PMID- 9405933 TI - Using codon usage to predict genes origin: is the Escherichia coli outer membrane a patchwork of products from different genomes? AB - Analysis of the codon usage of genes coding for the structural components of the outer membrane in Escherichia coli, is consistent with the requirement for high expression of these genes. Because porins (which constitute the major protein component of the outer membrane), and LPS (which constitute the major outermost constituent of the outer membrane), are synthesized from genes displaying widely different codon usage, it is possible to investigate the origin of the outer membrane. The analysis predicts that the outer membrane might originate from a genome other than the genome coding for the major part of the cell. Such a special origin would explain in structural terms, the likely lethality of porins if they were inadvertently inserted within the inner membrane, giving rise to the Gram-negative bacterial type, having an envelope comprising two membranes, instead of a single cytoplasmic membrane and a murein sacculus. PMID- 9405934 TI - Structure of the mutant fibrillin-1 gene in the tight skin (TSK) mouse. AB - Mice carrying the tight skin (TSK) mutation harbors a 3.0-kb genomic duplication (exons 17-40) of the fibrillin-1 gene (Fbn-1) located on band F of chromosome 2 as TSK mutation. We cloned and sequenced the mutated Fbn-1 gene, since it is believed to be responsible for TSK syndrome. Sequence analysis showed numerous amino acid differences in the 5' and 3' segments between the TSK mutation and wild-type fbn-1 gene, but any amino acid difference between the TSK mutation and C57BL/6 mice. (TSK and C57B1/6 mice are genetically similar, differing only by TSK mutation.) Four amino acid differences were observed between two copies of TSK's fbn-1 gene encoded by exons 17-40. Our results suggest that the majority of structural differences occurred in the N and C termini segments during strain divergence and only a few after the duplication event. PMID- 9405935 TI - Identification and characterization of a novel human phosphatidylinositol 4 kinase. AB - The extensive sequence homology that exists among the catalytic domains of phosphatidylinositol 3- and 4-kinases allowed us to clone a novel human gene encoding a putative phosphatidylinositol kinase, NPIK. Among other known phosphatidylinositol 3- and 4-kinases, NPIK was most closely related to yeast PIK1 phosphatidylinositol 4-kinase. Several forms of NPIK cDNAs were isolated, and expression of NPIK message was detected in a wide variety of tissues. Fluorescence in situ hybridization and radiation hybrid analyses assigned the NPIK gene to human chromosome 1. Recombinant NPIK protein catalyzed a conversion from phosphatidylinositol to phosphatidylinositol 4-phosphate. The catalytic activity of NPIK was augmented by Triton X-100, and was reduced in the presence of adenosine. Using green fluorescent protein system we determined that NPIK is localized in the cytoplasm. Taken together, the data suggest that NPIK may play a pivotal role in regulating the synthesis of phosphatidylinositol 4-phosphate at the site(s) accessible from cytoplasm. PMID- 9405936 TI - A 3-Mb sequence-ready contig map encompassing the multiple disease gene cluster on chromosome 11q13.1-q13.3. AB - Despite the presence of several human disease genes on chromosome 11q13, few of them have been molecularly cloned. Here, we report the construction of a contig map encompassing 11q13.1-q13.3 using bacteriophage P1 (P1), bacterial artificial chromosome (BAC), and P1-derived artificial chromosome (PAC). The contig map comprises 32 P1 clones, 27 BAC clones, 6 PAC clones, and 1 YAC clone and spans a 3-Mb region from D11S480 to D11S913. The map encompasses all the candidate loci of Bardet-Biedle syndrome type I (BBS1) and spinocerebellar ataxia type 5 (SCA5), one-third of the distal region for hereditary paraganglioma 2 (PGL2), and one third of the central region for insulin-dependent diabetes mellitus 4 (IDDM4). In the process of map construction, 61 new sequence-tagged site (STS) markers were developed from the Not I linking clones and the termini of clone inserts. We have also mapped 30 ESTs on this map. This contig map will facilitate the isolation of polymorphic markers for a more refined analysis of the disease gene region and identification of candidate genes by direct cDNA selection, as well as prediction of gene function from sequence information of these bacterial clones. PMID- 9405937 TI - Structural analysis of Arabidopsis thaliana chromosome 5. II. Sequence features of the regions of 1,044,062 bp covered by thirteen physically assigned P1 clones. AB - A total of 13 P1 clones, each containing a marker(s) specifically mapped on chromosome 5, were isolated from a P1 library of the Arabidopsis thaliana Columbia genome, and their nucleotide sequences were determined according to the shot gun based strategy and precisely located on the physical map of chromosome 5. The total length of the sequenced regions was 1,044,062 bp. Since we have previously reported the sequence of 1,621,245 bp by analysis of 20 non-redundant P1 clones, the total length of the sequences of chromosome 5 determined so far reached 2,665,307 bp. The regions sequenced in this study were analysed by comparison with the sequences in protein and EST databases and analysis with computer programs for gene modeling; a total of 225 potential protein-coding genes and/or gene segments with known or predicted functions were identified. The positions of exons which do not exhibit similarity to known genes were also predicted by computer-aided analysis. An average density of the genes and/or gene was 1 gene/4,640 bp. Introns were identified in approximately 84% of the potential genes, and the average number and length of the introns per gene were 5.3 and 184 bp, respectively. These sequence features are essentially identical to those for the previously sequenced regions. The transcription level of the predicted genes has been roughly monitored by counting the numbers of matched Arabidopsis ESTs. The sequence data and gene information are available through the World Wide Web at http:@www.kazusa.or.jp/arabi/. PMID- 9405938 TI - Complementary DNA cloning and chromosomal mapping of a novel phosphatidylinositol kinase gene. AB - A cDNA for a putative new member for phosphatidylinositol kinase family was cloned from an adult human whole brain cDNA library. The predicted translation product was composed of 961 amino acid residues and contained a sequence feature characteristic for lipid/protein kinases. The messenger RNA was ubiquitously expressed in various tissues, while relatively higher expression was observed in heart, skeletal muscle and testis. The chromosomal location of the gene was determined by fluorescence in situ hybridization and PCR-based analyses with both a human/rodent monochromosomal hybrid cell panel and a radiation hybrid mapping panel. PMID- 9405939 TI - Use of polymerase chain reaction and antibody tests in the diagnosis of vertically transmitted hepatitis C virus infection. AB - Data on patterns of polymerase chain reaction (PCR) and antibody test results in infants born to hepatitis C virus (HCV)-infected mothers were systematically reviewed to aid development of optimum testing schedules and diagnostic criteria for vertically exposed infants and to facilitate early identification of infected infants. Survival and cross-sectional analyses were used to estimate the timing of initial PCR positivity and subsequent PCR negativity in infected infants, and maternal antibody loss in uninfected infants was estimated as a weighted average of individual study findings. Of 74 eligible infants with strong evidence of HCV infection, an estimated 89% (90% confidence interval, 80-95%) were first PCR positive by 3 months of age, and less than 10% had subsequent PCR negativity attributable to intermittent viraemia or resolved infection in the first 18 months of life. The negative predictive value of PCR at 3 months of age was greater than 98% at an assumed rate of 5% vertical transmission, but as low as 88% at 25% transmission. The inclusion of 22 infants, each with a single PCR positive result, increased the estimated frequency of resolved infections but made little difference to other estimates. A minority of PCR-positive infants had periods of antibody negativity by second- or third-generation assays, and among 297 uninfected infants, maternal antibody was not detected beyond 18 months. Thus, the majority of infected infants may be persistently PCR positive from 3 months of age, and the negative predictive value of PCR at 3 months is generally high. However, poor repeatability of PCR, inadequate infant follow-up, and inclusion of postnatally infected infants limits interpretation of the pooled data. Further studies using standardised PCR methodologies are needed. PMID- 9405941 TI - False-negative results of a ligase chain reaction assay to detect Chlamydia trachomatis due to inhibitors in urine. AB - The aim of this study was to assess the presence of inhibitors in urine specimens causing false-negative results in a commercial Chlamydia trachomatis gap-filling ligase chain reaction (Gap-LCR) assay. On testing of urine samples by the Gap-LCR assay and urethral swab specimens by cell culture, 73 (19%) Chlamydia trachomatis positive subjects were detected among 382 men attending a clinic for sexually transmitted diseases. In 56 subjects, the agent was detected in both the urine and the urethral samples, while 309 subjects were negative in both tests. In seven subjects urine samples were Gap-LCR positive (confirmed by a different Gap LCR assay), but the corresponding urethral swab samples were cell culture negative. In another ten subjects the urethral swab samples were cell culture positive, but their urine samples were Gap-LCR negative. Subsequent re-analysis of the urine samples including the addition of external Chlamydia trachomatis DNA indicated full or partial inhibition in nine of the cell culture-positive Gap-LCR negative subjects. When urine preparations were freeze-thawed and diluted prior to testing, Chlamydia trachomatis was detected in six of the ten initially Gap LCR-negative samples. Gap-LCR inhibitors were present in at least nine (12%) of the 73 urine preparations from the Chlamydia trachomatis positive individuals. Identification of samples containing Gap-LCR inhibitors and subsequent processing to reduce the inhibition increased the sensitivity of the test from 86% to 95%. PMID- 9405940 TI - Chlamydia pneumoniae respiratory infections among patients infected with the human immunodeficiency virus. AB - Thirteen cases of Chlamydia pneumoniae infection in patients seropositive for the human immunodeficiency virus (HIV) are described. The occurrence, the clinical spectrum, and the significance of the infection during HIV disease are compared with data reported in the literature. Chlamydia pneumoniae infection was established by a serologic micro-immunofluorescence test using standard diagnostic criteria. In four cases the results of serological tests were confirmed by direct immunofluorescence on respiratory specimens. Five patients developed focal pneumonia but recovered completely after specific antibiotic treatment. Three patients developed severe and diffuse interstitial pulmonary involvement, two of whom died of acute respiratory failure. Five patients developed upper respiratory tract infection. Using 39 pair-matched HIV seropositive subjects as controls, the cases of infection were found to be significantly associated with a previously diagnosed pulmonary disease. Upon retrospective analysis of 319 consecutive cases of pneumonia among HIV-infected patients, Chlamydia pneumoniae was the sole agent detected in eight (2.5%) cases, and Chlamydia pneumoniae together with other infectious agents was detected in seven (2.2%) cases. Chlamydia pneumoniae is a possible cause of severe respiratory infection in Italian HIV-infected immunocompromised patients, and its presence must be suspected when patients do not respond to therapy with beta lactam agents or to anti-Pneumocystis carinii treatment. PMID- 9405942 TI - Efficacy of different methods for detection of low Cryptosporidium parvum oocyst numbers or antigen concentrations in stool specimens. AB - The detection of Cryptosporidium parvum oocysts in stool specimens by acid-fast (AF) stains or immunofluorescence assays (IFA) requires the presence of large numbers of oocysts. To determine whether new commercially available enzyme immunoassays (EIAs) are more sensitive alternatives, three EIAs, a direct IFA, and the modified cold Kinyoun AF stain were compared, particularly with respect to detection of low oocyst numbers or antigen concentrations. Thirty-one negative and 31 calf stool-enriched human stool specimens were tested. One EIA method detected only nine positive specimens, demonstrating a sensitivity significantly less (p < 0.0001) than that of the IFA, the AF stain, and the other two EIAs. No differences could be found with respect to specificity. In addition, serial dilutions of 28 patients' stool samples containing cryptosporidian oocysts were prepared and examined using two EIAs, IFA, and the AF stain. One EIA yielded significantly inferior results (p < 0.0001), whereas the other one and the two microscopic methods did not differ significantly in either part of the study. The results indicate that the new EIAs do not exhibit higher sensitivities for detection of Cryptosporidium parvum than the two routinely used microscopic methods. Thus, for most laboratories, the IFA or AF stain may still represent the preferred method for the diagnosis of cryptosporidiosis. PMID- 9405943 TI - Risk factors, clinical features and outcome of Acinetobacter bacteremia in adults. AB - The medical records of 39 patients with Acinetobacter bacteremia identified in the period between 1985 and 1995 were reviewed. In 24 cases (62%) the bacteremia was considered to have been clinically significant. Most of the infections (79%) were nosocomial, and the majority of these were acquired in an intensive care unit. Ten (42%) patients developed septic shock complicating the bacteremia and 13 (54%) died. In most of these cases (85%), Acinetobacter bacteremia was thought to have caused or contributed to death. The following variables were associated with a greater risk of mortality: age > 65 years (OR = 16; p = 0.01); development of septic shock (OR = 22; p = 0.004); and the presence of coagulopathy (OR = 20; p = 0.03). PMID- 9405944 TI - Molecular typing of Acinetobacter baumannii from ten different intensive care units of a university hospital. AB - Thirty-one isolates of Acinetobacter baumannii were collected from ten intensive care units of an Austrian university hospital. All isolates were typed by enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC PCR). Two strains colonizing 13 infants in the neonatal intensive care unit were identified by ERIC-PCR. All other Acinetobacter baumannii isolates had highly divergent ERIC-PCR patterns, despite having the same antibiogram. Thus, a hospital-wide clonal distribution, as suggested by identical antibiogram patterns, was excluded by ERIC-PCR. PMID- 9405945 TI - Urinary tract infections in primary biliary cirrhosis and other chronic liver diseases. AB - In a study to determine the prevalence of urinary tract infections (UTI) in primary biliary cirrhosis, midstream specimens of urine from 97 females with primary biliary cirrhosis and 85 females with other chronic liver diseases were investigated prospectively for urinary pathogens and Mycobacterium gordonae. No significant differences between primary biliary cirrhosis and the two groups were observed in the prevalence of significant bacteriuria (11.3% vs. 7.1%), the prevalence of Escherichia coli UTI (9.3% vs. 7.1%) or the colony morphology of Escherichia coli. No mycobacterial species were grown from any sample. In both groups, the prevalence of UTI was higher in patients with cirrhosis (20% in both) than in those without. PMID- 9405946 TI - Longitudinal study of Aspergillus fumigatus strains isolated from cystic fibrosis patients. AB - The colonization over time of cystic fibrosis patients by Aspergillus fumigatus was investigated using a DNA fingerprinting method. Aspergillus fumigatus isolates collected sequentially for more than one year from six patients with cystic fibrosis were typed by Southern blot hybridization with a repetitive DNA sequence. Each cystic fibrosis patient harbored several strains of Aspergillus fumigatus that were isolated recurrently over time. Isolates collected from a cystic fibrosis patient with aspergilloma displayed the same genotype, suggesting that the infection was due to a single strain. Continuous isolation of the same genotype in another cystic fibrosis patient, however, was not correlated clinically with an Aspergillus infection. PMID- 9405947 TI - Analysis of isolates recovered from multiple sites of the nasopharynx of children colonized by nontypeable Haemophilus influenzae. AB - To determine whether the nasopharynx of children is colonized by a single or multiple strains of nontypeable Haemophilus influenzae, cultures were obtained from six nasopharyngeal sites in five children. For each child, all isolates yielded identical polymerase chain reaction fingerprints. The results indicate that these children were colonized in the nasopharynx with a single strain of nontypeable Haemophilus influenzae at one time. PMID- 9405948 TI - Colonization rates and serotypes of group B streptococci isolated from pregnant women in a Korean tertiary hospital. AB - In a study designed to provide data on the rates of maternal carriage of group B streptococci (GBS) in Korean women, vaginal, anorectal, and urethral swab specimens from 459 pregnant women and ear canal and umbilicus swabs from their 288 neonates were cultured with new Granada medium and selective Todd-Hewitt broth. Additionally, the serotypes of 64 isolates of GBS and the minimal inhibitory concentrations of seven antimicrobial agents for these isolates were determined. The rate of colonization by GBS in pregnant women and in their babies was 5.9% (27/459) and 0.7% (2/288), respectively. The rates of resistance of GBS isolated from pregnant women were 13.3% to clindamycin, 5% to erythromycin, and 98.3% to tetracycline. The majority of GBS isolates from pregnant women belonged to serotypes Ib (48.3%), Ia (24.1%), and III (20.7%). PMID- 9405949 TI - Pseudo-outbreak of listeriosis elucidated by pulsed-field gel electrophoresis. AB - Listeria monocytogenes was isolated from three patients hospitalized in two departments of the same hospital over a two-week period. A nosocomial outbreak of listeriosis was suspected. Patients presented with tenosynovitis, central venous catheter infection, and bacteremia. The first patient had a community-acquired infection, the second a nosocomial infection, and the source of the third patient's illness was uncertain. Epidemiological investigations failed to identify a common source of contamination within the hospital. The three strains were nontypeable by phage typing, but Smal macrorestriction analysis and pulsed field gel electrophoresis yielded three distinct profiles. Therefore, the three cases seemed to represent a cluster of sporadic cases as opposed to an outbreak of listeriosis. Rapid typing of isolates is essential in the early investigation of potential outbreaks of listeriosis and may prevent the initiation of expensive and time-consuming epidemiological investigations. PMID- 9405950 TI - Low prevalence of active parvovirus B19 infection in HIV-infected patients. AB - In a prospective study to determine the prevalence of parvovirus B19 in adult patients positive for the human immunodeficiency virus, a series of 55 consecutively presenting patients was tested for antibodies to parvovirus B19 and parvovirus B19 DNA. Specific IgG antibodies were detected in 96% of cases; specific IgM antibodies were present in 10%. Viral DNA was not detected in any of the sera analysed by polymerase chain reaction. Anemia could not be correlated with the presence of either IgG or IgM antibodies to parvovirus B19. Thus, no evidence of acute or chronic parvovirus B19 infection was found in the 55 patients. These findings suggest that active parvovirus B19 infection is uncommon in HIV-infected patients. PMID- 9405951 TI - Immunoglobulin G avidity in the serodiagnosis of congenital rubella syndrome. AB - The avidity of specific IgG was investigated in three infants with serologically verified congenital rubella infection. Two sera were taken from each infant: the first soon after birth and the second at the age of 23 to 31 months. Avidity of specific IgG was measured by a protein-denaturing enzyme immunoassay using urea as the elution factor, and avidity then determined by the end-point ratio (derived from antibody titration) and the avidity index methods. Rubella-specific IgM was present in the first sera of all patients, but not in the second sera. However, low avidity of specific IgG persisted in two children until age 23 to 31 months, as determined by the end-point ratio method. These data are in agreement with the findings of previous studies of avidity in congenital rubella, and show the usefulness of the protein-denaturing IgG-avidity assays employing the end point ratio method for serological diagnosis of congenital rubella even after disappearance of specific IgM. PMID- 9405952 TI - In vitro activity of the new ketolide HMR 3004 compared to an azalide and macrolides against Streptococcus pneumoniae and Haemophilus influenzae. AB - The purpose of this study was to compare the in vitro activity of a new ketolide, HMR 3004 (RU64004), to that of three macrolides and one azalide against 608 Streptococcus pneumoniae and 202 Haemophilus influenzae. Macrolide-resistant pneumococci were susceptible to HMR 3004, even if they were resistant to clindamycin. Against Haemophilus influenzae, HMR 3004 and azithromycin were nearly identical in potency; the macrolides were 8- to 16-fold less active. HMR 3004 may be useful for treating respiratory tract infections if sufficient concentrations can be achieved at the local sites of infection. PMID- 9405953 TI - Human T-cell lymphotropic virus infection in pregnant women in Spain. PMID- 9405954 TI - Pulmonary coinfection by Salmonella enteritidis and Pneumocystis carinii in a patient with the acquired immunodeficiency syndrome. PMID- 9405955 TI - Evaluation of a commercial serological kit for detection of salivary immunoglobulin G to Helicobacter pylori. PMID- 9405956 TI - The need for long-term treatment of depression. PMID- 9405957 TI - Advantages and limitations of the concept of antidepressant therapy. AB - The current availability of a range of effective treatments make it more than ever necessary for depression to be recognised. The emphasis of treatment has moved from acute episodes to the longitudinal course of affective disorders. In addition to Major Depressive Disorder, the need for treatment is now established in less severe variants, particularly Dysthymia; the same drugs are applicable, but should be part of a global strategy. In general, antidepressants have to be considered in relation to non-pharmacological treatment methods, of which the most important is ECT. A great variety of new drugs have been found to have antidepressant efficacy, but pharmacologically this action is not a specific or exclusive one. Side-effects have on the whole not been studied for long enough, or in depressed patients rather than healthy volunteers; there are marked variations between individuals in their reactions to different drugs. Co morbidity is a frequent issue in the treatment of depression, particularly for the elderly. Since overall suicide rates have not fallen in many countries, the treatment particularly of Major Depressive Disorder and Bipolar Disorder needs to be strengthened. PMID- 9405958 TI - Prevention of stress-induced morphological and cognitive consequences. AB - Atrophy and dysfunction of the human hippocampus is a feature of aging in some individuals, and this dysfunction predicts later dementia. There is reason to believe that adrenal glucocorticoids may contribute to these changes, since the elevations of glucocorticoids in Cushing's syndrome and during normal aging are associated with atrophy of the entire hippocampal formation in humans and are linked to deficits in short-term verbal memory. We have developed a model of stress-induced atrophy of the hippocampus of rats at the cellular level, and we have been investigating underlying mechanisms in search of agents that will block the atrophy. Repeated restraint stress in rats for 3 weeks causes changes in the hippocampal formation that include suppression of 5-HT1A receptor binding and atrophy of dendrites of CA3 pyramidal neurons, as well as impairment of initial learning of a radial arm maze task. Because serotonin is released by stressors and may play a role in the actions of stress on nerve cells, we investigated the actions of agents that facilitate or inhibit serotonin reuptake. Tianeptine is known to enhance serotonin uptake, and we compared it with fluoxetine, an inhibitor of 5-HT reuptake, as well as with desipramine. Tianeptine treatment (10 mg/kg/day) prevented the stress-induced atrophy of dendrites of CA3 pycamidal neurons, whereas neither fluoxetine (10 mg/kg/day) nor desipramine (10 mg/kg/day) had any effect. Tianeptine treatment also prevented the stress-induced impairment of radial maze learning. Because corticosterone- and stress-induced atrophy of CA3 dendrites is also blocked by phenytoin, an inhibitor of excitatory amino acid release and actions, these results suggest that serotonin released by stress or corticosterone may interact pre- or post-synaptically with glutamate released by stress or corticosterone, and that the final common path may involve interactive effects between serotonin and glutamate receptors on the dendrites of CA3 neurons innervated by mossy fibers from the dentate gyrus. We discuss the implications of these findings for treating cognitive impairments and the risk for dementia in the elderly. PMID- 9405959 TI - Antidepressant efficacy in the treatment of dysthymia. AB - Although the existence of chronic depression has been recognized for a long time, their definition was too inaccurate to enable reliable studies concerning their treatment. Among the numerous diagnostic classes that patients with chronic depression were assigned to, neurotic depression was the most common one, and was often considered to be unresponsive to antidepressant medication. Since DSM-III introduced 'Dysthymic Disorder', a new research was developed on the efficacy of tricyclic antidepressants, MAOIs or new antidepressants, whose results reversed previous opinion on its unresponsiveness. However the interpretation of those studies are hampered by methodological problems, yet unresolved, pertaining to the difficulty to differentiate dysthymia and major depression, to the frequency of 'double-depression', the usual mildness of symptoms in dysthymia, and the need of long-term trials. PMID- 9405960 TI - Experimental findings in the study of the reduction of alcohol intake. AB - Alcohol dependence represents a major problem in public health and different animal models of dependence have been developed in rodents with the aim of studying the mechanisms of alcohol abuse. Different ways of animal alcoholisation have been established. They permit a better understanding of which neurotransmitter system is involved in the regulation of alcohol dependence. Considerable attention has been given to the role of serotonin in the control of both alcohol craving and alcohol related pathologies, i.e. anxiety, aggression or memory loss. In conclusion, the use of animal models of alcohol abuse facilitates the understanding of alcohol behavior and permits the development of new therapeutic agents. PMID- 9405961 TI - Efficacy of tianeptine in major depressive disorders with or without melancholia. AB - The efficacy of tianeptine in the treatment of major depressive episodes was assessed in three double-blind placebo-controlled studies. In a first double blind study comparing tianeptine (37.5 mg/day) with placebo, 126 patients with Major Depression or a Depressed Bipolar Disorder were treated for 42 days; 60% of these patients fulfilled DSM-III-R criteria for melancholia. Final MADRS scores showed the efficacy of tianeptine in comparison with placebo (P = 0.007). This result was confirmed by the time course of the Severity of Illness (CGI item 1) (P = 0.015). 58% of the patients responded to tianeptine versus 41% to placebo. In another study comparing tianeptine (37.5 mg/day), imipramine (150 mg/day), and placebo, 186 depressed patients were treated for 42 days. The patients had either Major Depression or Depressed Bipolar Disorder, without melancholia (DSM III-R). In the intention-to-treat analysis, final MADRS scores showed a better efficacy of tianeptine and imipramine than placebo (P = 0.012 and P = 0.034, respectively). There were 56% responders on tianeptine vs 48% on imipramine, and 32% on placebo. A third study involved 244 patients with Major Depression with or without melancholia (DSM-III-R). They were treated in a parallel group design with tianeptine (37.5 mg/day) or tianeptine (75 mg/day) or placebo for 42 days. The high rate of placebo-responders (> 65%) did not allow any conclusion about the efficacy of tianeptine. Altogether, tianeptine was shown to be an effective and safe medication for the treatment of major depressive episodes. However, a controlled study in endogenous depression would be useful to determine the position of tianeptine among the other antidepressants in this indication. PMID- 9405962 TI - Tianeptine and alcohol dependence. AB - Several arguments are in favour of the use of antidepressant drugs in alcohol dependent patients, especially those acting on the serotoninergic system: (1) neurochemical data indicate the interaction between alcohol and 5-HT metabolism, (2) pharmacological studies show an improvement in the behaviour of alcoholized animals treated with antidepressants, (3) depression is a frequent disease in alcoholic patients. Tianeptine has been shown to be active in the treatment of depression in patients with history of alcohol abuse or dependence. In a first double-blind study performed versus amitryptiline, depression after withdrawal was improved by tianeptine, and biological abnormalities usually related to chronic alcohol intake tended to decrease. Similar results were found in an open study carried out on 277 alcoholic patients treated for 1 year. As these patients were depressed, no definite conclusion could be drawn from these results in respect of a specific action of tianeptine on alcohol dependence. Thus, a multicentre double-blind study has been performed which compared tianeptine (12.5 mg t.i.d) and placebo in 342 non-depressed patients fulfilling DSM-III-R criteria for Psychoactive Substance Dependence (alcohol). Other inclusion criteria were: daily alcohol intake higher than 80 g, minimum score of 3 on the Short-Mast Questionnaire, mean corpuscular volume above 98 fl and/or gamma Gt more than twice the upper limit of normal. The patients were treated for 9 months. The intention-to-treat population and the per protocol population were made up of 327 patients and 111 patients, respectively. The main efficacy criterion was the absence of alcoholic relapse (abstinence) defined by the patient's statements, the investigators clinical judgement and some biological parameters: alcohol blood levels, gamma Gt levels. Secondary criteria were the evolution of the alcohol consumption in the patients who relapsed, cumulative abstinence duration, a visual analogue scale for the evaluation of the appetence for alcohol and the clinical global impressions scale. The statistical analysis showed no difference between both groups in respect of the maintenance of abstinence (intention-to treat and per protocol populations). In spite of the methodological problems of the studies in dependence (choice of the inclusion and efficacy criteria, especially), the preliminary results obtained with the serotoninergic antidepressants were not confirmed in the different trials performed in the maintenance of alcohol abstinence. The indication of tianeptine should be restricted to the treatment of depressive syndromes, which have a high lifetime prevalence in the alcoholic patient, and which have a noticeable role on the alcoholic relapse. PMID- 9405963 TI - Observations on initial cell loss after intraportal hepatocyte transplantation of 5'-bromo-deoxy-uridine-labeled hepatocytes. AB - In the present study, attempts are made to improve posttransplant survival of intraportally transplanted hepatocytes in a syngeneic hepatocyte transplantation (HTX) model. Engrafted hepatocytes were detected and quantified after pretransplant labeling with 5'-bromo-deoxy-uridine. In order to enhance the survival percentage, three mechanisms that possibly influence cell survival were manipulated: (1) administration of Matrigel (extracellular-matrix components, Matrigel Basement Membrane Complex, Micronic BV, Lelystad, The Netherlands) in order to improve the environmental conditions of the transplanted hepatocytes; (2) immunosuppression of the recipients by 5 Gy total body irradiation and cyclosporin-A administration (25 mg/kg, 3 x weekly), and (3) Kupffer cell depletion by injection of dichloromethylene-diphosphonate-filled liposomes. The results were analyzed in relation to HTX without treatment. Cell survival was approximately 14% (10 days after HTX), and the transplanted hepatocytes were distributed equally throughout the various recipient liver lobes. However, no increase was attained by Matrigel coadministration, immunosuppression, or Kupffer cell depletion. PMID- 9405964 TI - Orthotopic rat liver transplantation and bile duct reconstruction by a splint technique. AB - The aim of the present study is to investigate the impact of bile duct reconstruction by a splint technique, a method which has not been sufficiently researched in animals after liver transplantation. Three experimental groups were set up: I = control, sham operation; II = bile duct reconstruction; III = orthotopic rat liver transplantation (ORLT). After bile duct reconstruction, serum levels of ASAT and ALAT in group II revealed a peak on the first postoperative day. The transplanted animals (group III) showed a second peak in liver enzyme levels on the fifth postoperative day; it was significantly higher than in group II. Serum bilirubin was more elevated in the transplant group, with a peak on day 7. Morphological investigations at the end of surgery revealed only intralobular necrosis and reactive changes in the liver capsule (group II); after transplantation (group III), there was also interstitial and intracellular edema, fatty degeneration and disintegration of the sinusoidal lining. One month later, necrosis, bile duct proliferation, cholestasis, cholangitis and vascular alterations were found in groups II and III. Furthermore, an increased rate of hepatocellular and bile duct proliferation was observed. These findings are partly due to the bile duct reconstruction. We recommend that a bile duct reconstruction control group should be included in ORLT experiments. PMID- 9405966 TI - Effect of dibutyryl cyclic adenosine monophosphate on skeletal muscle reperfusion injury in the rat. AB - To clarify the action of dibutyryl cyclic adenosine monophosphate (DBcAMP) on reperfused ischemic muscle, experiments were conducted using the hindlimbs of 18 male Lewis rats. At the midportion of the thigh all tissues except for the femoral artery and vein were transected, and a route for continuous intravenous infusion was secured in a contralateral limb vein. After inducing total ischemia by clamping the femoral artery and vein with a vascular clamp for 4 h, the limb was reperfused for 1 h. Blood flow was then compared using the hydrogen gas clearance method in a group in which DBcAMP 10 mg was continuously infused from a vein in the contralateral hindlimb from 1 h prior to the induction of ischemia to 1 h after the completion of ischemia (DBcAMP group), a group in which saline was infused in the same manner (control group), and a group which was subjected to biopsy alone (biopsy group). The percent change in blood flow was significantly higher in the DBcAMP group than in the control group at 15 and 30 min after the release of the clamp. Adenosine triphosphate (ATP), phosphocreatine (PCr), and lipid peroxide (LPO) were measured in tissue samples obtained 1 h after reperfusion. Serum LPO was measured in blood samples collected at the same time. ATP values were higher in the DBcAMP group than in the control group. PCr was significantly higher in the DBcAMP group than in the control group. LPO levels in skeletal muscle tissue did not differ significantly between the DBcAMP and control groups. In contrast, serum LPO levels were significantly lower in the DBcAMP group than the control group. On morphologic analysis the control and DBcAMP groups showed normal vascular endothelial cells and absence of the 'no reflow' phenomenon. These data confirm that in this reperfusion model the administration of DBcAMP enhances the viability of skeletal muscle cells. Moreover, mediated by an effect on vascular endothelial cells this agent is thought to be of help in mitigating the vascular endothelial cell injury occurring in acute ischemic injury. DBcAMP may be a useful agent in mitigating skeletal muscle ischemia-reperfusion injury. PMID- 9405965 TI - Allopurinol reduced hepatic ischemia-reperfusion injury exacerbated by inhalation of high-concentration oxygen in rats. AB - Exposure to high-concentration oxygen (O2) increases lipid peroxidation of the cellular membrane, leading to tissue injury which may involve hepatic ischemia reperfusion (I/R) injury. We examined the effects of inhaling high-concentration O2 on hepatic I/R injury with allopurinol, which is a xanthine oxidase inhibitor. Partial hepatic ischemia was performed in rats with or without allopurinol under 21 or 100% O2 inhalation. Levels of lipid peroxide, serum liver enzymes, and hepatocellular oxidative stress in the 100% O2 group were significantly higher than in the 21% O2. Administration of allopurinol significantly inhibited those changes in the 100% O2 group. Severe degeneration of mitochondria were noted in the 100% O2 group, but appeared to be reduced by allopurinol. Results suggest that inhalation of high-concentration O2 during liver surgery may increase lipid peroxidation and exacerbate hepatic I/R injury, but those changes may be prevented by allopurinol. PMID- 9405967 TI - Epidermal growth factor induces increased mucosal thickness of the small intestine in mouse. AB - Epidermal growth factor (EGF) is known to exert a mitogenic effect in different tissues, including the digestive tract. The aim of the present study was to evaluate whether long-term infusion of EGF causes trophic effects in the gastrointestinal tract of female mice. The animals were infused subcutaneously in the neck with human recombinant EGF in a dose of 10 micrograms/kg/h (1.6 nmol/kg/h) using an osmotic minipump for 1, 3 and 7 days, respectively. Tritiated thymidine was continuously infused intraperitoneally during the same period, except in the 7-day group, where it was infused during the last 3 days. The mucosal thickness was measured microscopically. As a measurement of DNA synthesis, the amount of thymidine retained in the mucosa was registered using a scintillation counter. After 1 day of EGF infusion, the mucosal thickness was increased in the antrum and, after 3 days, in the fundus. In the proximal duodenum, an increased depth of the crypts was seen after 1 day, followed by increased villi height after 3 and 7 days; in the distal duodenum, EGF evoked increased villi height after 3 and 7 days. The height of villi was increased after 7 days in the jejunum and ileum in the EGF-treated animals. The tritium incorporation was increased in the fundus of the stomach and the proximal duodenum in the EGF-treated animals after 3 days, whereas no significant increase in tritiated thymidine incorporation could be detected in the EGF-treated animals after 1 and 7 days compared to the controls. In conclusion, continuous infusion of EGF evoked increased mucosal thickness in the small intestine, while the trophic effects were only of a short duration in the stomach and absent in the colon. PMID- 9405969 TI - Immune monitoring of surgical colorectal malignancies through carcinoembryonic antigen and alpha-fetoprotein serum levels, flow-cytometric blood lymphocyte phenotyping and tumor DNA analysis. AB - 67 patients with operable colorectal adenocarcinomas were divided into two groups based on criteria referring to DNA flow-cytometric models and the histopathological type and stage of intraoperatively harvested tumor samples: low aggressive tumor group and highly aggressive tumor group. Two kinds of immunological markers were investigated: humoral (carcinoembryonic antigen and alpha-fetoprotein serum levels were measured by ELISA) and cellular (blood lymphocyte counts of the following subpopulations: CD3+, CD4+, CD8+/CD11b+, CD19+, CD16+/CD56+, HLA-DR+/CD3+, measured by means of flow cytometry). All assays were similarly performed before and 2, 4, 6 and 8 weeks after tumor exeresis. Results seem to reveal a relationship between patients without immunological recovering and a bad postoperative clinical evolution of the highly aggressive colorectal malignant tumors. The significance of multiway immune monitoring of colorectal cancer is discussed. PMID- 9405968 TI - Quantitative monitoring of blood supply to knee joint transplants in dogs. AB - BACKGROUND: Transplantation of vascularized knee joints has become technically feasible, but graft rejection as well as failures of the vascular anastomoses remain critical hazards. We therefore tested the potential of repetitive non invasive duplex sonography to detect changes of the arterial blood flow following canine knee joint transplantation. METHODS: Four transplantations and, as controls, 4 replantations of intact canine knee joints were performed. The follow up was 6 months. During this period, repetitive duplex sonography measurements as well as tests of knee joint function were performed. Six months postoperatively, angiographies were performed and all joints were explanted for histological investigation. RESULTS: The luminal diameters of the implanted popliteal artery remained constant in the transplanted animals (preop. 2.6 +/- 0.2 mm, 6 months postop. 2.7 +/- 0.2 mm) but decreased in the autografted controls (preop. 2.9 +/- 0.3 mm, postop. 2.0 +/- 0.3 mm). The time-averaged velocity of the popliteal artery blood flow decreased in both groups 1 month postoperatively. Subsequently, blood flow velocity recovered in transplanted animals but remained low in replanted controls. Significant arterial wall thickening was also detected in transplanted animals as compared to controls. Six months postoperatively, hypervascularization of transplanted joints was confirmed by angiography and thickening of the arterial wall by histology. Furthermore, histology identified mild to chronic allograft rejection in all transplanted joints in spite of controlled cyclosporin A trough level immunosuppression. CONCLUSIONS: Chronic rejection of transplanted vascularized knee joints appears to be associated with vessel wall thickening and hypervascularization rather than with vascular rarefaction (picture of the 'arbre mort') that is characteristic of the rejection of most parenchymatous organs. Duplex sonography appears to be sensitive in detecting the corresponding changes of blood supply. PMID- 9405970 TI - Implantation on the suture material and efficacy of povidone-iodine solution. AB - Suture implantation of viable exfoliated tumour cells may be responsible for local recurrence of colorectal cancer. Using a colon cancer cell line, we obtained a suture implantation without intraperitoneal metastasis in about 80% of the control animals, when sacrificed on the 2nd postoperative week. The cytotoxic efficacy of povidone-iodine (PVP-I) was tested in vivo by a rat model with viable intracaecal tumour cells, and in vitro by trypan blue exclusion and the MTT assay. In vivo PVP-I at 5% significantly reduced the incidence of tumour growth, while the product at 2.5% had a significant effect in only the monofilament polypropylene group. In an in vitro toxicity study, PVP-I higher than 0.16% was effective at killing almost all tumour cells. PVP-I had effective cytotoxicity in vivo and in vitro, being less cytotoxic in vivo than in vitro. PMID- 9405971 TI - The computer-assisted localizer, a navigational help in microneurosurgery. AB - The computer-assisted-localizer (CAL) achieves a direct linkage between preoperative radiological images and individual intraoperative anatomical findings. Experiences with our system demonstrate that CAL improves the intraoperative orientation and facilitates the neurosurgical procedure. The system described here consists of a mechanical articulated robot arm with six degrees of freedom and a three-dimensional image processor. After calibration, the displayed image dynamically pointed out the exact intraoperative localization in three perpendicular sectional views. Meanwhile, CAL was successfully used in 73 selected microneurosurgical procedures. PMID- 9405972 TI - Gaseous neurotransmitters in the hypothalamus. The roles of nitric oxide and carbon monoxide in neuroendocrinology. PMID- 9405973 TI - Interleukin-1 beta inhibits in vitro pulsatile progesterone secretion and stimulates prostaglandin F2 alpha secretion by micro-retrodialyzed baboon corpus luteum. AB - Interleukin-1 beta (IL-1 beta) modulates steroidogenesis and prostaglandin (PG) secretion by dispersed luteal cells of some non-primate species. To determine if IL-1 beta affects progesterone (P) and PGF2 alpha secretion by the baboon corpus luteum (CL), we microretrodialyzed the intact CL for 48 h; 12 h media (baseline), 12 h with IL-1 beta (3 IU/h), 12 h media only (post- IL-1 beta) and 12 h with cAMP (5 nmol/h). Four CL from the midluteal phase (LH + 8 days) were studied. P was measured by a sensitive and specific radioimmunoassay and PGF2 alpha by an enzyme immunoassay in the 10-min fractions of retrodialysates collected. P secretions were analyzed for peaks by PC Pulsar (3.0) and total P retrieved/12 h for each experimental segment was calculated. P secretion was pulsatile. Pulses of P declined from 8.2 +/- 1.2/12 h (mean +/- SEM) before to 5.0 +/- 1.2 h after IL-1 beta treatment (P = 0.022), but increased to 10.2 +/- 4.3/12 h with cAMP. Interpulse interval increased significantly from 92 +/- 23 min (baseline) to 137 +/- 31 min (p = 0.025) after IL-1 beta treatment. Total P secreted decreased significantly from 2471 +/- 515 nmol/12 h (baseline) to 1480 +/- 167 nmoles/12 h during IL-1 beta and 788 +/- 85 nmoles/12 h after IL-1 beta (P = 0.015). P was immediately suppressed after starting IL-1 beta in 2 CL but declined only towards the end of treatment in the other 2 CL. PGF2 alpha secretion increased during IL 1 beta with a further increase after IL-1 beta, while P secretion was progressively inhibited. Therefore, IL-1 beta is luteolytic to the primate midluteal phase CL by inhibiting P while simultaneously stimulating PGF2 alpha secretion, demonstrating paracrine-autocrine interaction within the luteal tissue. PMID- 9405974 TI - Cilostazol, a cAMP phosphodiesterase inhibitor, attenuates the production of monocyte chemoattractant protein-1 in response to tumor necrosis factor-alpha in vascular endothelial cells. AB - The induction of monocyte chemoattractant protein-1 (MCP-1) in vascular endothelial cells is thought to be an initial event in the development of atherosclerotic lesions. Therefore, inhibition of MCP-1 production may exhibit some effects in preventing atherosclerosis. In the present study, we found that 10 microM cilostazol, a cAMP phosphodiesterase inhibitor, increased the intracellular cAMP content by a twenty-five times of the basal level and resulted in the reduction of basal MCP-1 release by 41% from 168 +/- 11 ng/24 hr/mg protein to 99 +/- 14 ng/24 hr/mg protein (P < 0.001) from cultured human umbilical vein endothelial cells. Furthermore, 10 microM cilostazol also significantly attenuated the dose-dependent increment of MCP-1 production by tumor necrosis factor-alpha. The inhibition was consistent with the reduction of MCP-1 mRNA level, possibly through reduced activation of transcription factor NF kappa B level. Similarly, 1 mM dibutyryl cAMP inhibited MCP-1 production in endothelial cells. These data suggest that cilostazol inhibits MCP-1 production through increased intracellular cAMP levels and modulation of its expression in vascular endothelial cells. PMID- 9405975 TI - Defective stimulation of thyroxine 5'-deiodinase activity by cold exposure and norepinephrine in brown adipose tissue of monosodium glutamate-obese mice. AB - In order to examine the possible change in the thyroid hormone metabolism in the monosodium glutamate (MSG)-obese mice, we determined the iodothyronine deiodinase activity of brown adipose tissue (BAT), liver and kidney of male and female mice. There was no significant difference in the type II thyroxine 5'-deiodinase (T4 5'DII) activity in BAT between MSG-obese and control mice when they were kept at the ambient temperature of 22 degrees C. T4 5'DII activity in BAT of control mice increased markedly after exposure to cold (4 degrees C) for 4 h; however, the extent of cold-induced increase in T4 5'DII activity in BAT of MSG-obese mice was greatly reduced. Injection of norepinephrine (NE) (0.8 mg/kg, s.c.) 4 h previously increased T4 5'DII activity in BAT of control mice, but NE-induced increase in T4 5'DII activity was also markedly reduced in BAT of MSG-obese mice. Both cold- and NE-induced increase in T4 5'DII activity was greater in female, although similar tendency was obtained in male mice. Type I 3,3',5' triiodothyronine deiodinase (rT3 5'DI) activity of liver and kidney, and serum thyroxine (T4) and 3,5,3'-triiodothyronine (T3) levels in MSG-obese mice were essentially the same as those of the control male and female mice irrespective of cold exposure. These results suggest that defective stimulation of T4 5'DII activity of BAT by cold in the MSG-obese mice is due to deficient sympathetic input to BAT and/or to diminished response of BAT to NE, and may contribute to a possible cause of inability of MSG-obese mice to maintain body temperature under cold exposure. PMID- 9405976 TI - Antigonadotrophic effect of Mycobacterium tuberculosis. AB - The present investigation has been carried out to study if the Mycobacterium tuberculosis has any direct effect on granulosa cell progesterone production. Granulosa cells from pregnant mare serum gonadotropin (PMSG)-treated immature rats were incubated in replicates with and without the ultrasonicated Mycobacterium tuberculosis in the presence and absence of human chorionic gonadotrophin (hCG). Progesterone production and granulosa cell viability were assayed. The mycobacterial lysate significantly inhibited the basal production of progesterone. The lysate completely blocked the stimulatory effect of hCG. Any cytotoxic effect of the lysate was ruled out as none of the treatments decreased the viability of the granulosa cells as compared with the control values. PMID- 9405977 TI - Effect of estradiol on serum triglyceride lipoprotein levels and fatty acid composition in castrated rats. AB - Triglyceride content and fatty acid composition of rat serum lipoproteins showed specific variations after castration and estradiol treatment. Triglyceride levels decreased in VLDL after castration and in LDL and HDL after low doses of estradiol. High doses of estradiol enhanced triglyceride levels in VLDL and decreased them in LDL. Fatty acid composition showed a complex pattern: after castration, monoenoic acids decreased and essential fatty acids increased in all lipoprotein classes, enhancing the EFA/NEFA and EFA/ME ratios. Both doses of estrogen lowered these ratios in VLDL and LDL, but decreased them in HDL with high doses and enhanced them in HDL with low doses. PMID- 9405978 TI - Effect of macrophage colony stimulating factor on the advanced atherosclerosis in Watanabe heritable hyperlipidemic rabbits. AB - We have reported that macrophage colony-stimulating factor (M-CSF) prevents atherosclerosis in young WHHL rabbits (Atherosclerosis 93:245, 1993). In the present study, we injected recombinant human M-CSF (250 micrograms/day) into WHHL rabbits aged 11 months 3 times a week after advanced atherosclerosis was established. After 8 months of treatment, we did not find any significant difference in plasma lipid levels, cholesterol ester content in the aorta or macroscopic atherosclerosis lesion area between M-CSF treated and non-treated rabbits. There was, however, a significant difference in the ratio of intimal to medial thickness (1.08 vs 1.7, p < 0.01). Thus, M-CSF may influence vascular smooth muscle cell function and modify the process of atherosclerosis in advance lesions. PMID- 9405979 TI - Persistent GAD 65 antibodies in longstanding IDDM are not associated with residual beta-cell function, neuropathy or HLA-DR status. AB - Persistent humoral autoimmunity to the enzyme glutamic acid decarboxylase (GAD) has been described in a substantial proportion of patients with insulin-dependent diabetes mellitus (IDDM) of long duration. The source of the stimulus for this autoimmune reactivity is still unknown. Because the GAD 65 isoform is mainly expressed in pancreatic beta-cells and in the nervous system we investigated in the present study of the largest number of well characterized patients with longstanding IDDM (n = 105; median duration: 21 years; range: 10-46 years) the presence of autoantibodies to GAD 65 and their relationship to a residual C peptide response or peripheral and autonomic neuropathy. Additionally we studied the HLA-DR status relative to GAD 65 antibodies in 86 out of the 105 individuals. One hundred healthy control subjects and 100 recent onset IDDM patients were also studied for GAD 65 antibodies. GAD 65 antibodies were detected in a radioligand binding-assay with recombinant human GAD 65 and were present in 32% of the long term diabetic patients, 82% of the recent onset IDDM patients and in 3% of the healthy control subjects. A preserved C-peptide response to i.v. glucagon (Hendriksen criteria) was observed in 23% of the long-term IDDM patients. Autonomic neuropathy and peripheral neuropathy was identified using criteria based on both symptoms and formal testing giving a frequency of 67% vs 79%. The HLA specific DR 4/X was observed in 47% and HLA-DR 3/X in 22% of the long-term IDDM patients. Patients who were heterozygous for DR3/DR4 were found in 23% of the cases. GAD 65 antibodies were significantly less frequent in the long-term IDDM patients compared to recent onset IDDM (p < 0.001), and diabetes duration showed a significant negative correlation with GAD 65 antibody index levels (r = 0.22, p < 0.01). Interestingly, GAD 65 antibodies were not significantly correlated either with residual beta-cell function or neuropathy and no particular HLA-DR status was associated with persistent GAD 65 antibodies. In conclusion neither residual beta-cell function nor diabetic neuropathy or a certain HLA-DR specificity are exclusively associated with persistent autoimmunity directed to GAD 65 in longstanding IDDM. The stimulus for the persistent humoral immune response and its significance for the disease process and its complications remain to be established. PMID- 9405980 TI - Lack of effect of captopril on glomerular hyperfiltration in normoalbuminuric normotensive insulin-dependent diabetic patients. AB - The aim of the present study was to evaluate the effects of captopril on the glomerular filtration rate (GFR) and urinary albumin excretion rate (UAER) of normoalbuminuric normotensive insulin-dependent diabetes mellitus (IDDM) patients with and without glomerular hyperfiltration. Eleven normoalbuminuric (UAER < 30 micrograms/min) patients (age: 34.3 +/- 4.6 years: diabetes duration: 9.5 +/- 6.4 years) participated in the study. Six patients were considered to be hyperfiltering (GFR > or = 134 ml/min/ 1.73m2). GFR (51Cr-EDTA single injection technique), extracellular volume (ECV; distribution volume of 51Cr-EDTA), UAER (RIA) and metabolic and biochemical parameters were measured at baseline, after 6 weeks on captopril (25 mg p.o. twice daily) and after 6 weeks off captopril. Plasma renin activity (PRA; RIA), plasma aldosterone (RIA) and blood volume (51Cr red cell labeled) were measured at baseline and after 6 weeks on captopril. The baseline clinical and laboratory characteristics of hyperfiltering and normofiltering IDDM patients were similar. GFR did not change during the study (144.1 +/- 28.8; 139.7 +/- 21.8; 132.8 +/- 29.9 ml/min/1.73 m2) either in patients with hyperfiltration (164.6 +/- 20.7; 153.8 +/- 18.3; 148.6 +/- 31.0 ml/min/1.73 m2; n = 6) or without hyperfiltration (119.6 +/- 11.1; 123.2 +/- 11.9; 113.8 +/- 14.4 ml/min/1.73 m2; n = 5). Also, ECV (22.2 +/- 3.6; 21.5 +/- 4.3; 21.5 +/- 3.5 L/1.73 m2), UAER (3.9 [0.4-22.1]; 4.0 [0.2-11.4]; 3.7 [2.0 26.2] micrograms/min), systolic (112 +/- 13; 105 +/- 10; 111 +/- 11 mmHg) and diastolic (76 +/- 12; 72 +/- 9; 73 +/- 12 mmHg) blood pressure did not change. No difference in blood volume (60.8 +/- 10.4; 62.3 +/- 8.4 ml/kg) or plasma aldosterone (10.4 +/- 4.9; 7.7 +/- 3.8 ng/dl) was observed between baseline values and values after captopril use. PRA increased (2.4 [0.4-22.1]; 12.9 [2.2 41.1]ng/ml/h) at the end of 6 weeks on captopril (P = 0.002). Fasting plasma glucose, glycated hemoglobin, fructosamine, plasma cholesterol and potassium, 24 h urinary urea and sodium were similar during the study. These results were unchanged when patients with and without hyperfiltration were analyzed as separate groups. From baseline to the end of 6 weeks on captopril there was no correlation between change in GFR and change in glycated hemoglobin (r = 0.02, P = 0.96), systolic (r = 0.23; P = 0.49) and diastolic (r = -0.32, P = 0.32) blood pressure, urinary urea (r = 0.21; P = 0.53) and UAER (r = -0.16; P = 1.00). In conclusion, captopril has no effect on the GFR and UAER of normoalbuminuric normotensive IDDM patients irrespective of the presence of glomerular hyperfiltration. PMID- 9405981 TI - Renal metabolism of uric acid in type I insulin-dependent diabetic patients: relation to metabolic compensation. AB - Patients with insulin-dependent diabetes mellitus and poor glycemic control show hypouricemia with hyperuricosuria. In the present study, we have evaluated whether a good glycemic control influences the renal handling of uric acid. Sixteen patients (8 male, mean age 22.4 +/- 7.2 years) were studied under two situations, poor glycemic control (glycemia > 11 mmol/L and HbA1 c > 10%) and good glycemic control (glycemia < 6 mmol/L and HbA1 c < 8.5%). A group of 16 normal subjects served as the control group (8 male, mean age 21.9 +/- 9.1 years). In the poor glycemic control phase, patients showed lower plasma uric acid levels (0.18 +/- 0.06 mmol/L) and higher fractional urinary excretion of uric acid (16.1 +/- 9.3%) than the controls (0.28 +/- 0.06 and 8.2 +/- 1.9%, respectively). When a good glycemic control was reached, plasma uric acid increased (0.22 +/- 0.05), but it was still lower than that of the controls and fractional excretion of UA was normalized. Plasma uric acid was inversely correlated to glycemia (r = -0.34, p < 0.05) and to HbA1 c (r = -0.56, p < 0.0008) and fractional excretion of uric acid was directly correlated to glycemia (r = 0.39, p < 0.03) and HbA1 c (r = 0.73, p < 0.00005). These results indicate that the hypouricemia and hyperuricosuria of insulin-dependent diabetes mellitus is corrected by an adequate glycemic control, suggesting that these alterations are of a functional origin and due to a defective metabolic control. PMID- 9405982 TI - Dietary regulation of the very low density lipoprotein receptor in mouse heart and fat. AB - The very low density lipoprotein (VLDL) receptor is a member of the LDL receptor family. As opposed to the LDL receptor, the VLDL receptor is expressed primarily in muscle and adipose tissue. Although the VLDL receptor is capable of binding lipoproteins, its functional role is still unclear. Previous studies found that VLDL receptor expression is unaffected by fasting in the rat. The current studies examined whether VLDL receptor expression is altered with fasting in the mouse. Balb/c mice were fasted for periods up to 48 hours, killed, hearts and epididymal fat obtained, and total membranes prepared. To detect the VLDL receptor a portion of the rat VLDL receptor was expressed as a bacterial fusion protein, purified and used to immunize rabbits. The antibodies raised specifically recognized intact VLDL receptor. When cardiac membranes were immunoblotted, VLDL receptor expression increased progressively with fasting, doubling at 36 hours. In contrast, VLDL receptor expression decreased progressively with fasting in membranes from epididymal fat, being reduced 70% by 48 hours. Thus, VLDL receptor expression appears to be regulated in mouse heart and fat by nutritional perturbation, supporting a potential role for the VLDL receptor in the delivery of triglycerides/fatty acids as fuel. PMID- 9405983 TI - Cardiac secretion of adrenomedullin by percutaneous transluminal coronary angioplasty. PMID- 9405984 TI - Influence of lead on type-I iodothyronine 5'-monodeiodinase activity in male mouse. PMID- 9405985 TI - Management of dry retinal folds. AB - BACKGROUND: To describe the surgical technique used in the management of two cases of dry retinal folds which occurred following retinal detachment surgery. METHODS: Two patients with persistent dry retinal folds. RESULTS: Patients underwent complete vitrectomy and membrane peeling, followed by injection of perfluorocarbon liquid. The retina was prodded and massaged flat with a silicone tipped cannula under the perfluorocarbon liquid. The folds disappeared and the retina remained flat. CONCLUSION: The use of gentle manipulation under heavy perfluorocarbon liquids is an effective strategy for managing longstanding dry posterior retinal folds. PMID- 9405986 TI - Indocyanine green angiography in central serous chorioretinopathy. ICG angiography in CSC. AB - PURPOSE: To analyse images obtained by indocyanine green angiography in central serous chorioretinopathy (CSC). METHODS: Ninety patients affected with CSC were examined using indocyanine green angiography. RESULTS: CSC was detected in 127 of the 180 eyes examined. Leakage points were detected in 99 eyes with fluorescein angiography; in 85 of these eyes, they corresponded to hyperfluorescence with indocyanine green angiography, while a hyperfluorescence of the neuroepithelial detachment was seen in 21 eyes. Areas of choroidal hyperpermeability were seen in all 127 eyes with CSC and in 9 fellow eyes. With ICG angiography, the appearance of pigment epithelial detachments was similar to that previously described (early hyperfluorescence and later hypofluorescence), and was seen in 47 eyes. In 103 eyes, hypofluorescent lesions of various sizes, were detected which became more marked in the later stages. These lesions corresponded to retinal pigment epithelium lesions in fluorescein angiography, mainly hyperfluorescence caused by window defect. We were also able to observe RPE atrophic tracts in 31 eyes. These tracts appeared hyperfluorescent in 11 eyes where a minimal amount of RPE atrophy was present and hypofluorescent in 20 eyes in which the tract had marked RPE atrophy. CONCLUSION: The results obtained confirm the finding of choroidal hyperpermeability and subretinal diffusion of ICG, which indicate involvement of the choroid in CSC. The observation of progressively hypofluorescent lesions corresponding to retinal pigment epithelium alterations suggests that there may be as yet unknown interactions of pigment epithelium and ICG. PMID- 9405987 TI - Heparin and heparin-surface-modification reduce Staphylococcus epidermidis adhesion to intraocular lenses. AB - Bacterial adherence to intraocular lenses (IOLs) could be the cause of endophthalmitis following cataract surgery and lens implantation. The majority of cases of postoperative endophthalmitis are caused by microflora that reside on or near the eye of the patient. Staphylococcus epidermidis commonly colonizes the eyelid margin and conjunctiva and is the most common organism causing postoperative endophthalmitis. In this study, the in vitro adherence of S. epidermidis to regular poly-methyl methacrylate (PMMA) IOLs and to heparin surface-modified (HSM) PMMA IOLs was investigated. The effects of heparin and antibiotics in solution on the adherence of bacteria to regular PMMA IOLs were evaluated. Adhesion of bacterial cells to IOLs was determined by counting the viable cells attached to the lenses. Significantly, fewer S. epidermidis attached to HSM-PMMA IOLs and to regular PMMA IOLs treated with heparin than to PMMA IOLs (p < 0.001). Furthermore, bacteria attached in significantly lower numbers to regular PMMA IOLs treated with heparin than to HSM-PMMA IOLs (p = 0.0031). Antibiotics in solution had no significant effect on bacterial adherence to PMMA IOLs. These data indicate that the use of HSM-PMMA IOLs and treatment of PMMA IOLs with heparin could diminish the incidence of postoperative endophthalmitis and intraocular inflammation associated with IOL implantation. PMID- 9405988 TI - Presumed multifocal cryptococcol choroidopathy prior to specific systemic manifestation. AB - PURPOSE: Disseminated cryptococcosis is a major cause of morbidity and mortality in immunocompromised individuals, especially those with the acquired immunodeficiency syndrome (AIDS). Early diagnosis and treatment greatly improves the outcome, so clinical clues that lead to prompt diagnosis are important. METHODS: Three patients with AIDS in whom multifocal choroiditis and choroidal lesions were the initial signs of disseminated cryptococcosis were treated with systemic amphotericin B and flucytosine. All of the patients had a systemic work up that included evaluation of the cerebral spinal fluid (CSF). RESULTS: All three patients who were seen with the choroidal lesions as the presenting sign were noted to have either positive titers for cryptococcus or cultures that grew cryptococcus in the CSF. The choroidal lesions are presumed to be due to cryptococcus as no histopathologic or microscopic studies were available for ocular tissues. The choroidal lesions started to resolve one to three months after systemic treatment with amphotericin B and flucytosine. CONCLUSION: Primary choroidal lesions in patients with AIDS may herald severe systemic disseminated disease. Funduscopic examination, however, may detect disseminated cryptococcal disease before other overt clinical manifestations, thereby allowing prompt institution of effective therapy. PMID- 9405989 TI - Indocyanine green angiographic findings in fellow eyes of patients with unilateral occult neovascular age-related macular degeneration. AB - PURPOSE: To determine the role of indocyanine green angiography (ICGA) in the prediction of the development of choroidal neovascularization (CNV). METHODS: We reviewed the ICG angiograms of 124 patients with newly diagnosed unilateral occult CNV secondary to age-related macular degeneration (AMD) in whom the fellow eye had only drusen by biomicroscopy and fluorescein angiography. Follow-up data of at least 12 months were obtained in all eyes. RESULTS: 113 fellow eyes had a normal ICG study, while 11 fellow eyes revealed plaque-like late hyperfluorescence. During an average follow-up time of 18.2 months exudative AMD developed in 7 of 11 eyes with late hyperfluorescence on ICGA and in 6 of 113 with a normal ICGA study. CONCLUSION: ICGA may help to predict which fellow eyes are at higher risk of developing future exudative changes. PMID- 9405990 TI - Surgical closure of macular hole using an absorbable macular plug. AB - BACKGROUND: The surgical management of macular holes has been a subject of controversy in recent years. Various techniques such as vitrectomy, membrane peeling, and gas tamponade with or without transforming growth factor-beta 2, and recently the use of autologous platelets have produced closure rates from 58% to 96%, depending on the stage of the hole. METHODS: The authors present preliminary results in a study of 19 consecutive patients with stage 3 or stage 4 macular hole who underwent vitrectomy followed by placement of an absorbable partially cross-linked gelatin plug in the macular hole. The vitreous cavity was filled with a nonexpanding gas or air alone; the patient was instructed to maintain prone positioning for 2-3 days. RESULTS: Anatomic attachment of the edges of the macular hole was achieved in 19 out of 19 patients with a minimum follow-up period of 6 months (average 11.5 months). CONCLUSIONS: A cross-linked gelatin plug can effectively reattach the edges of macular holes of stages 3 and 4. Its use is recommended only in macular holes in high myopes with posterior staphyloma or recurrent macular hole. PMID- 9405991 TI - Cholesterol granuloma of the orbit--pathogenesis and surgical management. AB - BACKGROUND: Cholesterol granuloma of the orbit is a rare entity, and its pathogenesis is still poorly understood. We report on 6 cases including one patient who had been examined by X-ray prior to the tumor's clinical manifestation. PATIENT DATA: All tumors were located in the superior temporal orbit. Histologically, they revealed the typical features of a cholesterol granuloma without any epithelial elements. They infiltrated the bone but left the soft tissues largely intact. Complete surgical removal of the granulomatous mass was attempted in each case, and particular attention was given to thorough abrasion of the bone. Only 2 patients reported a previous trauma. In one of them, retrospective evaluation of the X-ray scan taken a few hours after his accident revealed no definite changes in the orbital bone at the site of the future tumor. The only recurrence developed in a patient in whom the bony base of the tumor had not been drilled out completely. CONCLUSIONS: The origin of orbital cholesterol granuloma remains unknown. According to our data and those available in the literature, trauma is not a precondition but may accelerate growth. Some non epithelial malformation in the bone, with a predelection in the temporal upper quadrant, might be the origin. To prevent a recurrence it appears essential to totally erase the tumor from its bony bed. PMID- 9405992 TI - Macular-threatening traction detachment of the retina in diabetic proliferative retinopathy, treated by laser. AB - PURPOSE: To treat patients with traction detachment of the central retina by laser in order to avoid vitrectomy. METHODS: Focal treatment of and around the flat detached retinal area with the argon laser. RESULTS: Seven patients were treated for macular-threatening traction detachment of the retina. The non rhegmatogenous traction detachment was flat and circumscribed. Panretinal photocoagulation (PRP) with the argon laser was performed prior to treatment of the traction in three of the cases, in four it was carried out in addition to PRP. In each of the seven patients partial or complete reattachment and stabilization for many years was achieved, rendering vitrectomy unnecessary (mean follow-up after therapy: 40.1 months). In no case was a laser-induced hole produced, and in no patient did the visual acuity decrease. CONCLUSION: The favourable results following photocoagulation can be explained by the tight retinal/choroidal scar formation (laserpexy). Laserpexy is only recommended for eyes with slight initial detachment. PMID- 9405993 TI - The European Ophthalmic Pathology Society. A brief history. PMID- 9405994 TI - Human bladder urine oxygen content: implications for urinary tract diseases. AB - Urine dissolved oxygen (DO) was measured in 40 healthy subjects and 115 patients divided into 4 groups according to their disease. Group 1 (20 patients) had lower urinary tract infection (UI), Group 2 (30 patients) had urinary stone disease (USD), Group 3 consisted of 50 end-stage chronic renal failure patients (CRF) and 15 patients in Group 4 were affected by influenza viral infection (IVI). Urinary and arterial PO2, PCO2 and pH were also measured in 20 healthy subjects. The other 20 healthy volunteers were subjected to submaximal exercise and afterwards urinary DO was estimated. Results revealed that in healthy subjects urinary DO or PO2 is not correlated with urinary pH or arterial pH, PO2 and PCO2. Also, urinary DO did not significantly vary on consecutive days. Urinary DO reflects mainly the renal metabolic state, being increased in conditions of decreased kidney metabolism such as CRF. Submaximal physical exercise, fever or urinary tract infection may significantly reduce urinary DO, whereas DO remains unaffected in uncomplicated USD. Human urinary DO is related to serum creatinine and urine volume. Our results indicate that urinary DO may be a useful indicator in clinical practice. PMID- 9405995 TI - Renal agenesis, ureteral ectopia into seminal vesicle, vas deferens agenesis and hemivertebra: an incomplete form of caudal regression syndrome? AB - The association of seminal vesicle cyst and upper urinary tract malformation is well known in the literature [1]. More rarely, urogenital malformations are associated with vertebral [2] or anorectal anomalies [3]. A 35-year-old infertile man with unilateral renal and deferential agenesis, seminal vesicle cyst and hemivertebra is reported. This complex malformative syndrome has been reported previously by Sheih et al. [4] and, to our knowledge, this is the third case described in the literature. PMID- 9405996 TI - Spontaneous nephrocutaneous fistula: a rare complication of reflux nephropathy. AB - A rare case of nephrocutaneous fistula due to reflux nephropathy in a 4-year-old female child is presented here. No such case has been described previously in the English literature. PMID- 9405997 TI - Should we get routine urothelial biopsies in every stone surgery? AB - Physical traumas have been implicated as intrinsic risk factors for the progression of urothelial tumours. In stone disease, histologic changes of the urothelium have a wide spectrum. We want to show the importance of biopsies for identification of these changes. In this study, we investigated the histologic changes of the urothelium in stone patients. There were 16 squamous metaplasia, 14 pyelitis follicularis, 5 pyelitis or ureteritis cystica, 4 polypoid pyelitis or ureteritis, 2 encrusted pyelitis in 43 stone patients, and 5 calcium, 2 long standing struvite and 2 mixed calculi histories were found in 9 of 14 upper urothelial tumour patients. According to our results, it is important to identify the histologic changes of the upper urothelium during stone surgery for possible neoplastic progression in the future. If any suspicious finding is demonstrated, the patients should be enrolled in a follow-up programme or should be transferred to tumour treatment programme. Therefore, we propose to take biopsies of the urothelium in every stone surgery. PMID- 9405998 TI - Determination of in vitro drug sensitivity to a panel of cystostatic drugs and interferon alpha-2b in patients with renal cell carcinoma. AB - Fresh operative cells from 27 renal cell carcinomas (RCC) were cultured in vitro for the determination of in vitro drug sensitivity. Two samples were not culturable. Incubation was carried out in triplicate in the presence and absence of various concentrations of chemotherapeutic agents. Sensitivity of the tumour cells to interferon-alpha (IFN-alpha), cisplatin (CDDP), mitomycin C (MMC), vinblastine (VBL), doxorubicin (DOX), etoposide (ETOP), bleomycin (BLM), vincristine (VCR) were tested by a colorimetric assay using MTT. A preexposure viability over 75% was essential for in vitro drug sensitivity assay (IVDSA). Sensitivity was determined by a more than 50 +/- 2 SD% reduction from the control absorbance. All eight drugs in their low concentrations exhibited cell proliferation inhibition in 0-12% of RCCs. On the other hand, IFN-alpha in its higher concentration (60 IU/ml) was effective in 88% of RCCs. After IFN, CDDP was found to be the second most effective drug in its higher concentration (36% efficacy). The results indicate that IFN appears to be the most effective in vitro agent in our 25 RCCs and the clinical trials either as a monotherapy or multiple combinations of various agents should include IFNs for the treatment of RCC. PMID- 9405999 TI - Serum levels of some acute phase proteins in kidney and urinary tract urothelial cancers. AB - Early diagnosis of kidney and urothelial cancer requires some new sensitive and specific methods. In this study the diagnostic use of serum alpha 1-acid glycoprotein (alpha 1-AG), coeruloplasmin, alpha 1-antitrypsin (alpha 1-AT), alpha 2-macroglobulin (alpha 2-MG) and albumin in patients with kidney, urinary bladder and upper tract urothelial cancer was evaluated. In kidney cancer patients the serum levels of alpha 1-AG, coeruloplasmin and alpha 1-AT were significantly increased over the controls (p < 0.001), however, albumin was decreased (p < 0.005). Sensitivity was relatively high for alpha 1-AG (85%), albumin (85%) and alpha 1-AT (77%). In patients with urinary tract urothelial cancer alpha 1-AG, alpha 1-AT and coeruloplasmin were also increased but not as much as in kidney cancer. Sensitivity of alpha 1-AG (63%), albumin (75%) and alpha 1-AT (66%) was also lower than in kidney cancer. This study has established the relative importance of alpha 1-AT and albumin determination in patients with kidney as well as with urothelial cancer. PMID- 9406001 TI - Primary osteosarcoma of the urinary bladder. AB - Primary osteosarcoma of the bladder, together with transitional cell carcinoma, occurred in a 74-year-old female presenting with haematuria and lumbago. She was treated with chemotherapy and radical cystectomy. Though it was conceived to be curative, she died of a recurrence 5 months after the operation. Primary osteosarcoma of the bladder is an unusual and highly malignant disease. In spite of radical therapy, the outcome has been dismal with some exceptions. PMID- 9406000 TI - Urinary tract endometriosis: report of 2 cases and a review of the literature. AB - Endometriosis of the urinary tract is a relatively rare condition. Since clinical signs are not specific the diagnosis is difficult and the therapy is not well defined. Two cases and a review of the literature are presented. PMID- 9406002 TI - The American Urological Association Symptom Index: early postoperative evaluation of irritative and obstructive symptoms due to benign prostatic hyperplasia. AB - In order to evaluate the acute effects of two different treatments on changes in the American Urological Association symptom score, we divided 23 men with benign prostatic hyperplasia into 2 groups. Group 1 (n = 16) and group 2 (n = 7) were treated with transurethral resection of the prostate and visual laser ablation of the prostate, respectively. Twice before and about 1 week after surgery, patients completed the AUA symptom questionnaire and underwent urodynamic evaluation. The symptom indexes were subcategorized as obstructive and irritative symptoms. All symptom scores were identical in groups 1 and 2 preoperatively. Postoperatively, significant improvement was found in obstructive scores, the total score, maximum and average flow rates only in group 1. This outcome is probably the reflection of an essential dissmilarity in both therapies. Clinically, the obstructive subscore appears reactive to changes in obstruction and seems meaningful in follow-up even in the early postoperative days. PMID- 9406003 TI - Significance of examination of prostate-specific antigen and prostate-specific antigen density in patients with prostatic hyperplasia and prostate cancer. AB - Authors investigated PSA concentration and preoperative prostate volume in 113 histologically proved BPH and 31 prostatic cancer patients. PSA concentration was measured with the Hybritech kit, the prostate volume by ultrasound with the help of an ellipse and calculated by computer. There was no correlation between the age of patients and volume of the prostate, whereas a correlation was proved between marker concentration and prostate volume (p < 0.0001). The difference obtained by the correlation of the prostate volume and the PSA concentration ratio between the BPH and 31 tumorous patients (PSA density) was highly significant (0.143 vs. 0.699, p < 0.001). The use of PSAD further improves the diagnostic value of PSA. PMID- 9406004 TI - Painless treatment of hydrocele: EMLA cream anaesthesia and fibrin adhesive sclerotherapy. AB - Sclerotherapy for hydroceles was performed in 18 patients. Cutaneous anaesthesia was induced with an anaesthetic cream (lidocaine and prilocaine, EMLA cream) and a fibrin sealant (Tissucol) was injected into the sac after fluid aspiration. Patients experienced no pain during needle insertion and sclerosant procedure; 2 recurrences were observed during follow-up. EMLA cream anaesthesia and fibrin adhesive sclerotherapy represent a useful alternative to surgical treatment of hydroceles. PMID- 9406005 TI - Malakoplakia of the testis. AB - Malakoplakia of the testis presenting as painless enlargement of the testis in an 80-year-old man is described. The literature is reviewed. PMID- 9406006 TI - Peyronie's disease: intralesional treatment with interferon alpha-2A and evaluation of the results by magnetic resonance imaging. AB - In this clinical study, to determine the therapeutic efficacy of interferon (IFN) treatment for Peyronie's disease, we applied interferon alpha-2A (IFN alpha-2A) intralesionally in the treatment of Peyronie plaques in 15 patients and results were evaluated by magnetic resonance imaging (MRI). Patients whose plaque sizes were 0.5 and 1 cm responded better to the treatment. There was about a 90% lessening in the sizes of the plaques of 1.5 cm, 83.3% of 2 cm, as the ones which were 0.5 cm and 1 cm disappeared completely after treatment. As a conclusion, the treatment of Peyronie's disease with IFN alpha-2A is effective and side effects are minimum. PMID- 9406007 TI - Chemiluminescence response of whole blood in patients undergoing urological operations. AB - Polymorphonuclear leukocytes (PMNs) are one of the most important components of the defence mechanisms against bacterial infection. The functions of PMNs are believed to be impaired in patients during the perioperative period. Bactericidal function of PMNs was investigated together with the luminol-dependent chemiluminescence (CL) reaction of whole blood in 23 patients, 12 undergoing open surgery and 11 undergoing endoscopic surgery. Blood samples were collected one day before surgery (day -1) and 2 hours (day 0), 24 hours (day 1) and 7 days (day 7) after surgery. Counts of whole white blood cells (WBCs), PMNs and lymphocytes were not different between the two surgery groups. CL responses in the open surgery group were increased on days 0, 1 and 7. In the endoscopic surgery group, CL response was increased on day 1, but not on day 0 or day 7. These results suggest that the PMN function during the perioperative period was not impaired, but increased just after surgery, mainly due to an increasing number of WBC caused by the surgical intervention. PMID- 9406008 TI - Influence of sex and chronic haemodialysis treatment on total, free and acyl carnitine concentrations in human serum. AB - The influence of sex and haemodialysis treatment on serum total, free and acyl carnitine concentrations in healthy controls and chronic renal failure patients has been investigated. Patients on regular haemodialysis treatment generally displayed significantly decreased serum carnitine levels. The mean predialysis serum carnitine levels were not significantly different from the mean healthy control values. However, after dialysis a significant decrease in serum carnitine levels was observed compared to the predialysis and healthy control values. Moreover, serum ratio of acylated to free carnitine was significantly higher after haemodialysis as compared to both healthy controls and predialysis patients. Sex-related changes in serum total, free and acyl carnitine levels and ratios of acylated to free carnitine have been observed in healthy controls and patients on chronic haemodialysis treatment. PMID- 9406010 TI - Dietary protein restriction in combination with angiotensin converting enzyme inhibitor improves insulin resistance in patients with chronic renal disease. AB - Insulin resistance (IR) and secondary hyperinsulinaemia are major risk factors of atherosclerosis and probably also of related glomerulosclerosis. Angiotensin converting enzyme inhibitors (ACEI), while improving IR in essential hypertension, do not improve it in patients with chronic renal disease. Thus, the combination of ACEI and low protein diet was evaluated. Thirty-eight patients with various kidney diseases and mild to moderate impairment of kidney function were included in the study. Thirteen of them suffered from IR. Their dietary protein intake was decreased from > or = 1.0 g/kg/d to 0.6-0.7 g/kg/d. Moreover, they were treated by ACEI enalapril at dosages of 2-10 mg/d depending on the absence/presence and severity of hypertension. The patients were followed for 8 months. No clinically relevant kidney disease progression (KDP) was found. IR patients improved remarkably. IR was examined by the oral glucose tolerance test and glucose, insulin and C-peptide determinations. Their increased plasma triglyceride, VLDL concentrations and proteinuria decreased, HDL concentration increased. Acid-base balance and anaemia did not change. It is concluded that protein restriction in combination with ACEI treatment improve IR and the associated dyslipoproteinaemia and proteinuria. PMID- 9406009 TI - The effect of dietary enrichment with fish-oil on urinary excretion of N-acetyl beta-D-glucosaminidase and renal function in proteinuric patients with primary glomerulopathies. AB - The rate of progression of renal disease depends on many factors including serum lipids and tubulo-interstitial injury. Aim of the study was to see whether fish oil therapy may affect serum lipids and NAG excretion with urine (a marker of tubular cell damage) in humans with renal disease. The effects of dietary fish oil fatty acids on the serum lipids, NAG urinary excretion and serum arachidonic acid concentration were examined in thirteen primary glomerulonephritic patients with proteinuria and normal renal function. The regular diet enriched with 1650 mg n-3 polyunsaturated fatty acids (18%: 20:5; n-3 EPA and 12%: 22:5; n-3 DHA) was ingested for three months. At the end of fish-oil enriched diet neither creatinine clearance nor urinary protein excretion changed significantly. But serum concentration of HDL and arachidonic acid increased (48.0 +/- 15 vs. 52.0 +/- 14; p < 0.05), (0.47 +/- 0.13 vs. 0.72 +/- 0.29; p < 0.01), respectively. Simultaneously urine NAG excretion and serum LDL decreased (11.2 +/- 7.1 vs. 10.3 +/- 7.3; p < 0.05), (163.0 +/- 57 vs. 149.0 +/- 51, p < 0.01), respectively. We presume that fish-oil supplementation may have a beneficial effect on renal tubular cells in humans and it could be linked with arachidonic acid metabolism. PMID- 9406011 TI - Ultrasound enhances gene expression of liposomal transfection. AB - RATIONALE AND OBJECTIVES: Cationic liposomes are under development as delivery agents for gene therapy. The authors studied the effect of ultrasound on gene expression in cell cultures during liposomal transfection experiments. METHODS: Cationic liposomes of dipalmitoylethylphosphocholine and dioleoylphosphatidylethanolamine were used to transfect cultured HeLa, NIH/3T3, and C127I cells with the chloramphenicol acetyl transferase (CAT) gene. A cell viability assay was performed on cultured HeLa cells that were exposed to varying durations (5 seconds or 30 seconds) and intensities of 1 MHz continuous-wave therapeutic ultrasound after transfection, and gene expression was measured 48 hours later. RESULTS: Cells survived 30 seconds or less at a power level of 0.5 watts/cm2 but died when exposed for 60 seconds or longer. Exposures of 5 seconds and 30 seconds of ultrasound resulted in significant increases in gene expression in all three cell types tested in this experiment. CONCLUSIONS: Relatively low levels of ultrasound energy can be used to enhance gene expression from liposomal transfection. Additional experiments are needed to optimize this process and clarify the mechanisms involved. PMID- 9406012 TI - Ultrasound-mediated destruction of contrast agents. Effect of ultrasound intensity, exposure, and frequency. AB - RATIONALE AND OBJECTIVES: Although ultrasound contrast microbubbles theoretically could serve as tracers for the noninvasive quantification of blood flow, results have been inconsistent. Accurate quantification may be limited by ultrasound energy-mediated microbubble destruction. This study examined the effect of different ultrasound delivery parameters on microbubble destruction. METHODS: Experiments were performed in an in vitro hydraulic perfusion model consisting of a thin-walled rubber tube encased in agar. Ultrasonic parameters tested during different parts of the experiment were (1) intensity, (2) duration, and (3) frequency. Four ultrasound contrast agents: Aerosomes MRX115 (ImaRx Pharmaceuticals Corp., Tucson, AZ), Imagent AF0150 US (Alliance Pharmaceutical Corp., San Diego, CA), Levovist (Berlex Laboratories, Wayne, NJ), and Echogen (Sonus Pharmaceuticals, Bothel, WA) were imaged with three different ultrasound systems: ATL Ultramark AM-9 HDI, Vingmed 800 and Hewlett-Packard 2500. RESULTS: Microbubble destruction and reductions in reflectivity were noted in all agents tested. Although no significant reductions in counts or reflectivity occurred at 0.3 W/cm2 with any agent, exposure to 25 W/cm2 produced more than 80% reductions in both microbubble counts (P < 0.0001) and reflectivity (P < 0.0001). Declines in reflectivity were increased by longer exposure to ultrasound (P < 0.0001); slower flow through an ultrasound beam (P < 0.0001); continuous, rather than intermittent, imaging (P = 0.0002); use of a higher pulse repetition rate (P < 0.0001); and exposure to 2.5 MHz, rather than 7.5 MHz, ultrasound (P < 0.0001). CONCLUSIONS: Ultrasound energy-mediated destruction of contrast microbubbles is a function of many factors, including ultrasound intensity, duration, and frequency. Optimization of ultrasound delivery parameters may be used to maximize or minimize the destruction of ultrasound contrast agents. PMID- 9406013 TI - Phase I clinical trials of MRX-115. A new ultrasound contrast agent. AB - RATIONALE AND OBJECTIVES: Stabilized microbubbles are under development as contrast agents for medical ultrasound. The authors report the results of Phase I clinical trials of a new ultrasound contrast agent based on lipidencapsulated perfluorocarbon gas microbubbles. METHODS: Lipids encapsulating perfluoropropane gas (Aerosomes MRX-115, ImaRx Pharmaceutical Corp., Tucson, AZ) were evaluated in Phase I clinical trials. Two separate studies were performed. The first was a single escalating-dose study (n = 30 subjects), and the second was a multiple dose study (n = 18 subjects) with rechallenge in several subjects (n = 4) after 21 days. Echocardiographic examinations were performed before and after contrast agent for each test drug administration for both studies, with the exception of the rechallenge group. Doses tested in the single-dose study ranged from 0.005 mL/kg to 0.100 mL/kg body weight. In the multiple-dose study, five doses of 0.005 mL/ kg to 0.030 mL/kg (0.025-0.150 mL/kg total dose) were evaluated. Studies were single-masked, placebo-controlled, and safety assessment and adverse events were monitored. RESULTS: All doses in both studies were well tolerated with no treatment-related changes in safety measures for either study. Left ventricular cavity and myocardial enhancement were seen with all doses of MRX-115. CONCLUSIONS: MRX-115 is a promising new intravascular ultrasound contrast agent that was safe and well tolerated at the doses evaluated in these studies. PMID- 9406014 TI - Preclinical evaluation of the pharmacokinetics, biodistribution, and elimination of MS-325, a blood pool agent for magnetic resonance imaging. AB - RATIONALE AND OBJECTIVES: The authors evaluate MS-325, a new albumin-targeted magnetic resonance imaging (MRI) contrast agent, for its pharmacokinetics, biodistribution, and elimination characteristics in multiple animal species. METHODS: Studies were performed in rats, rabbits, and nonhuman primates at intravenous doses ranging from 0.025 to 0.20 mmol/kg. Concentrations of MS-325 in blood, urine, feces, and organs were determined using gadolinium-153-labeled MS 325 and gamma counting or by using non-labeled MS-325 and inductively coupled plasma atomic emission spectrometry. RESULTS: In rabbits and nonhuman primates, MS-325 is approximately 85% to 95% bound to serum proteins and, as a result, exhibits low volume of distribution (Vd) values, 0.11 to 0.14 L/kg, and a long elimination half-life (Te1/2), 2 to 3 hours. Some dose-dependence in the parameters is apparent in rabbits. MS-325 is eliminated primarily through the renal system in non-human primates. In contrast, the behavior of MS-325 in rats is different, exhibiting increased biliary excretion, a larger Vd value, and a shorter Te1/2. CONCLUSIONS: The pharmacokinetics and elimination profile of MS 325, including vascular retention and renal excretion, are favorable for use in humans as an intravascular contrast agent for MRI. PMID- 9406015 TI - Targeting dendrimer-chelates to tumors and tumor cells expressing the high affinity folate receptor. AB - RATIONALE AND OBJECTIVES: The authors developed a new method for delivering contrast agents to tumors and tumor cells. Gadolinium complexes of folate conjugated dendrimer-chelates increased the longitudinal relaxation rate of tumor cells expressing the high-affinity folate receptor, hFR. The coupling of folate to polymeric chelates, composed of a dendrimer backbone, targets these chelates to endogenous folate binding proteins. These proteins exist in both the serum of patients with cancer and on the cell surface of many human cancers of epithelial origin. METHODS: The authors attached folic acid to a generation four ammonia core polyamidoamine dendrimer. The folate-dendrimer was reacted with 2-(4 isothiocyanatobenzyl)-6-methyl-diethylenetriaminepentaacetic acid to form the polymeric chelate f-PAMAM-TU-DTPA. For fluorescent studies, the generation four dendrimer was reacted with fluorescein-5-isothiocyanate and carboxytetramethylrhodamine succinimidyl ester, followed by capping the remaining amines with succinic anhydride. RESULTS: The study results show that cells accumulate the folate-conjugated dendrimer in a receptor specific manner. Tumor cells expressing the high-affinity folate receptor showed a 650% increase in the mean fluorescence. This increase occurred with a rapid rise to 325%, followed by a slow increase to 650%. It required both the expression of the hFR and the coupling of folic acid to the dendrimer. Excess free folic acid inhibited the binding of the folate conjugated polymer. Fluorescent microscopic study showed that the folate-conjugated dendrimer binds to the cell surface and is accumulated within the cells. Treatment of tumor cells that express the hFR with gadolinium complexes of the folate-conjugated polymeric chelate increases the longitudinal relaxation rate by 110%. This increase was inhibited by an excess of free folic acid. CONCLUSIONS: These data demonstrate that folate-conjugated magnetic resonance imaging contrast agents represent a promising new approach to tumor targeting. PMID- 9406016 TI - Hemodynamic tolerance of intravascular contrast agents for magnetic resonance imaging. AB - RATIONALE AND OBJECTIVES: Intravascular contrast agents for magnetic resonance imaging (MRI) facilitate the quantification of tissue perfusion. The authors determined the hemodynamic tolerance of these agents. METHODS: Doses of 0.05, 0.15, and 0.45 mmol/kg of the polymeric intravascular contrast agent gadolinium DTPA-polylysine, and di-nitrobenzyl-gadolinium-DTPA, a non-polymeric intravascular contrast agent with high protein binding, and gadolinium-DTPA dimeglumine, a paramagnetic contrast agent with extracellular distribution, were injected into 18 normal male rats as a peripheral intravenous bolus. Systolic, diastolic, and mean blood pressure, left ventricular end-diastolic and developed pressure, positive rate of pressure change (+dP/dt), dP/dt, the rate-pressure product, and heart rate were recorded during a period of 20 minutes. Hemodynamic effects were established by analysis of variance for repeated measurements. RESULTS: There was a transient increase of all blood pressure parameters and contractility for Gd-DTPA-polylysine at the dose of 0.45 mmol/kg only. Di nitrobenzyl-Gd-DTPA increased blood pressure parameters at 0.45 mmol/kg only. At doses of 0.05 and 0.15 mmol/kg, no significant hemodynamic effects were observed. CONCLUSIONS: The authors conclude that Gd-DTPA-polylysine is hemodynamically safe at doses to 0.15 mmol/kg and acts like a plasma expander at higher doses after peripheral bolus injection in normal rats. Additional investigations are indicated to elucidate the mechanism of a nonsignificant and satiable transient hemodynamic depression after injection of 0.05 mmol/kg DNB-Gd-DTPA. PMID- 9406017 TI - Relative blood volume measurements by magnetic resonance imaging facilitate detection of testicular torsion. AB - RATIONALE AND OBJECTIVES: The authors determine the utility of relative blood volume measurements (rBV) using a blood pool marker for magnetic resonance imaging (MRI) in detection of early testicular torsion. METHODS: Testicular torsion was induced in rats by counterclockwise 720 degrees rotation and fixation of the testis in the scrotum. MPEG-PL-DTPA-Gd enhanced MRI (30 mumol Gd/kg bolus injection) was performed 1 hour after torsion at 1.5 T using fat-suppressed three dimensional fast spoiled gradient-recalled sequence for relative blood volume measurement and three-dimensional time-of-flight sequence for MR angiography (MRA). RESULTS: The rBV of the torqued testes was significantly lower (13.3% +/- 13.5%) than that of testes with sham operation (97.7% +/- 5.3%; P < 0.05). Rats with testicular torsion showed larger regions of ischemia than did animals with sham operation (63.4% +/- 13.0% versus 4.0% +/- 2.8% of all pixels in testis; P < 0.01). The MRA of testicular torsion showed engorgement of the distal testicular vein as a sign of venous compression or total disappearance of the testicular vein, indicating arterial insufficiency. CONCLUSIONS: The authors conclude that MPEG-PL-DTPA-Gd can be used to obtain functional (rBV), morphologic (tunica enhancement), and angiographic (venous engorgement, arterial compromise) findings that should improve the diagnosis of testicular torsion in the acute setting. PMID- 9406018 TI - Magnetic resonance imaging-histomorphologic correlation studies on paramagnetic metalloporphyrins in rat models of necrosis. AB - RATIONALE AND OBJECTIVES: The authors intended to confirm previous findings that paramagnetic porphyrins are avid only for intratumoral nonviable tissues, but not for viable tumor cells, and to test the hypothesis that necrosis, regardless of location and origin, can be visualized by metalloporphyrin enhanced magnetic resonance imaging (MRI). METHODS: Intravenous administrations of gadolinium mesoporphyrin (Gd-MP), manganese tetraphenylporphyrin (Mn-TPP), manganese methylpyrroporphyrin-gadopentetate dimeglumine complex (Mn-MPP-Gd) and manganese tetra(4-sulfonatophenyl)porphyrin (MnTPPS4) at 0.05 mmol/kg were compared with those of gadopentetate dimeglumine (Gd-DTPA) at 0.1 mmol/kg in 38 rats with cholestatic liver necrosis, alcohol- and laser-induced coagulation necrosis in liver, and skeletal muscle, reperfused hepatic infarction, and segmental renal infarction. T1-weighted spin echo MRI (TR/TE = 300/15 mseconds) was acquired before and as long as 48 hours after injection, matched with histologic findings, and correlated with Gd/ Mn tissue content measurements. RESULTS: Both Gd-DTPA and the four metalloporphyrins initially caused a similar nonspecific negative contrast enhancement in the necrosis. However, a strong and persisting positive enhancement (necrosis-to-normal contrast ratio ranging from 1.5 to 2.0) developed only with metalloporphyrins in all types of necrosis. In liver and kidney, Gd and Mn concentrations at 24 hours were comparable in necrotic and normal tissues. In muscle, the concentrations were more than eight times higher in necrotic than in normal tissue. CONCLUSIONS: The implied affinity of metalloporphyrins for necrosis with presumably increased relaxivity suggests a possible mode of targetability for MRI contrast media that may elicit novel applications. PMID- 9406020 TI - Magnetic resonance imaging of pulmonary ventilation. Initial experiences with a gadolinium-DTPA-based aerosol. AB - RATIONALE AND OBJECTIVES: The authors investigate whether a modified gadolinium (Gd)-DTPA formulation can be aerosolized and used as a contrast agent for magnetic resonance (MR) ventilation imaging of the lungs. METHODS: Gadolinium DTPA (gadopentetate dimeglumine, Schering AG, Berlin, Germany, 100 mmol Gd/L) was modified by addition of mannitol (Sigma, Deisenhofen, Germany, 10 mg/mL) and the surface active detergent Lutrol F68 (BASF, Mannheim, Germany, 2 mg/mL). The imaging was performed in an anesthetized rat model after inhalation of the contrast agent aerosol (PulmoSonic, De Vilbiss, Germany, 10-minute nebulization). T1-weighted spin echo images (repetitive time [TR]/echo time [TE] = 40/3 mseconds) were acquired at 2 T (SIS 85; Sisco, Fremont, CA) before and as long as 120 minutes after administration of the contrast agent. RESULTS: The modified Gd DTPA aerosol elicited high and relatively homogeneous enhancement of the lung directly after nebulization. The enhancement was more pronounced than that obtained with a Gd-DTPA formulation without additives. CONCLUSIONS: Gadolinium DTPA-based aerosol appears to be a suitable contrast agent for MR ventilation imaging in an experimental animal model. Modification by mannitol (to increase proton density through a slight additional osmotic effect) and a detergent (to reduce droplet size by decreasing surface tension) is suitable and effective in increasing signal intensity compared with Gd-DTPA without modification. PMID- 9406019 TI - Gadolinium chelates with weak binding to serum proteins. A new class of high efficiency, general purpose contrast agents for magnetic resonance imaging. AB - RATIONALE AND OBJECTIVES: The authors assess the effect of weak protein binding on the efficacy of gadolinium chelates as contrast agents for magnetic resonance imaging (MRI). METHODS: Chelates with no (gadopentetate dimeglumine), weak (gadobenate dimeglumine), and strong (B-21326/7) protein binding were compared by in vitro MRI at 2T (spin echo [SE]: repetition time [TR]/echo time [TE] 350/8 mseconds) on solutions in 0.5 mM bovine serum albumin and in rat whole blood, and by in vivo MRI at 2T on rat models of brain tumors (SE TR/TE 350/10 mseconds) and of focal blood-brain barrier disruption (SE TR/TE 400/15 mseconds) after injection of MPP+. Relaxation rate enhancement in the blood of normal rabbits was measured in vivo after administration of contrast agents using IR-Snapshot FLASH. RESULTS: Signal intensity enhancement measured in vitro for whole rat blood 0.1 mM in gadobenate was 142% relative to the same concentration of gadopentetate. Peak signal intensity enhancement in brain tumors was 87% +/- 8% and 64% +/- 5% after 0.1 mmol/kg intravenous administration of gadobenate and gadopentetate, respectively; in MPP+ lesions, the peak signal intensity enhancement was 22% +/- 9%, 32% +/- 7%, and 64% +/- 14% after 0.2 mmol/kg intravenous of gadopentetate, gadobenate, and B-21326/7, respectively. In rabbits, the relaxation enhancement of blood 5 minutes after B-21326/7 and gadobenate administration was 323% and 182%, respectively, relative to the same dose (0.1 mmol/kg intravenous) of gadopentetate. CONCLUSIONS: Weak protein binding can substantially increase the efficacy of gadolinium chelates as general purpose contrast agents for MRI. PMID- 9406022 TI - Time and time again: the phylogeny of melatonin as a transducer of biological time. AB - The circadian secretion of melatonin is a critical component in circadian and seasonal rhythms in many vertebrate species. This hormone is produced by photoreceptors and cell types derived from photoreceptors in vertebrate retinas and pineal complexes via circadian regulation of the biosynthetic enzymes arylalkylamine N-acetyltransferase and hydroxyindole-O-methyltransferase at both transcriptional and posttranscriptional levels. The question of whether other multicellular animals and organisms from other taxa produce melatonin in a homologously regulated pathway is at this point unclear, but preliminary evidence suggests that vertebrate and insect melatonin are produced by convergent or parallel phylogenies. The existence and function of algal and plant melatonin is worthy of further study but is unresolved at this point. In vertebrates, the role of melatonin in behavioral and systems physiology follows two phylogenetic patterns. First, the circadian regulation of visual system structures, including the hypothalamic suprachiasmatic area, the inner retina, and retinorecipient and integrative visual structures, is a primitive characteristic among vertebrate species. Second, the relative loss of visual regulation and the presence of melatonin binding in the pars tuberalis of the adenohypophysis among mammals is a derived characteristic because these characteristics are present in this group only. PMID- 9406021 TI - A noninvasive method for monitoring renal status at bedside. AB - RATIONALE AND OBJECTIVES: The authors demonstrate the feasibility of monitoring renal status continuously and noninvasively at a patient's bedside, avoiding both radioactivity and blood and urine samples. METHODS: Gadolinium-153-labeled ProHance and a glomerular filtration rate (GFR) standard technetium-99m-DTPA were coadministered to anesthetized normal and nephrectomized rats with their tails hanging in a PC 20 spin analyzer. Blood samples and T1 measurements were collected and analyzed. RESULTS: Log time plots of 153Gd, 99mTc (from blood samples) and T1 of the rat tails were all linear and parallel. Halftimes were 32 +/- 2, 32 +/- 6, and 32 +/- 6 minutes for the decay of the T1, 153Gd and 99mTc, respectively. The halftime of the nephrectomized animal was 2000 +/- 4000 minutes. CONCLUSIONS: T1 of an appendage remote from the kidneys reflects the concentration of gadolinium in the blood, which is in rapid equilibrium with tissue interstitial space gadolinium. The decay in T1 of the appendage reflects glomerular filtration. Thus, it is feasible to detect changes in renal status at a patient's bedside by monitoring T1 of a finger or wrist using a small, inexpensive magnet. PMID- 9406023 TI - Melatonin: role in development. AB - Melatonin is the mammalian fetus's window to periodicity of the outside world. Through melatonin, the fetus "knows" what time of year it is and, in all likelihood, also knows the time of day. The best known function of melatonin during development is to communicate information about photoperiod and thereby adaptively regulate reproductive development. A second likely function of melatonin during development, which may be related to but more widespread than the first, is to entrain the developing circadian pacemaker. Prenatal maternal entrainment occurs in all of the eutherian mammals in which it has been examined, and in Syrian hamsters exogenous melatonin during development causes entrainment. The broader distribution and greater abundance of melatonin receptors during development, relative to mature animals, suggests that developmental effects of melatonin are greater and more diverse. The human fetal suprachiasmatic nucleus expresses melatonin binding sites and is therefore likely to be affected by both endogenous and exogenous melatonin with consequences for the prenatal and postnatal expression and entrainment of circadian rhythms. Caution is warranted, not only concerning the use of exogenous melatonin during pregnancy and lactation but also concerning behavior that might disrupt the mother's endogenous melatonin rhythm. PMID- 9406024 TI - Homeostatic versus circadian effects of melatonin on core body temperature in humans. AB - Evidence obtained in animals has suggested a link of the pineal gland and its hormone melatonin with the regulation of core body temperature (CBT). Depending on the species considered, melatonin intervenes in generating seasonal rhythms of daily torpor and hibernation, in heat stress tolerance, and in setting the CBT set point. In humans, the circadian rhythms of melatonin is strictly associated with that of CBT, the nocturnal decline of CBT being inversely related to the rise of melatonin. Whereas there is inconsistent evidence for the suggestion that the decline of CBT may prompt the release of melatonin, conversely, stringent data indicate that melatonin decreases CBT. Administration of melatonin during the day, when it is not normally secreted, decreases CBT by about 0.3 to 0.4 degree C, and suppression of melatonin at night enhances CBT by about the same magnitude. Accordingly, the nocturnal rise of melatonin contributes to the circadian amplitude of CBT. The mechanisms through which melatonin decreases CBT are unclear. It is known that melatonin enhances heat loss, but a reduction of heat production cannot be excluded. Besides actions on peripheral vessels aimed to favor heat loss, it is likely that the effect of melatonin to reduce CBT is exerted mainly in the hypothalamus, where thermoregulatory centers are located. Recent observations have shown that the acute thermoregulatory effects induced by melatonin and bright light are independent of their circadian phase-shifting effects. The effect of melatonin ultimately brings a saving of energy and is reduced in at least two physiological situations: aging and the luteal menstrual phase. In both conditions, melatonin does not exert its CBT-lowering effects. Whereas in older women this effect may represent an age-related alteration, in the luteal phase this modification may represent a mechanism of keeping CBT higher at night to promote a better embryo implantation and survival. PMID- 9406025 TI - Melatonin and seasonal rhythms. AB - The pineal hormone melatonin plays a ubiquitous role in biology as a chemical mediator of the effects of season on animal physiology and behavior. Seasonal changes in night length (scotoperiod) induce parallel changes in the duration of melatonin secretion (which occurs exclusively at night), so that it is longer in winter and shorter in summer. These changes in duration of nocturnal melatonin secretion, in turn, trigger seasonal changes in behavior. The retinohypothalamic pineal (RHP) axis's responses to light are highly conserved in humans. Like other animals, humans secrete melatonin exclusively at night, and they interrupt its secretion when they are exposed to light during the nocturnal period of its secretion. In many individuals, the RHP axis also is capable of detecting changes in the length of the night and making proportional adjustments in the duration of nocturnal melatonin secretion, producing the type of melatonin message that animals use to trigger seasonal changes in their behavior. This has been shown both in naturalistic studies in which melatonin profiles were compared in summer and winter and in experimental studies in which melatonin profiles were compared after chronic exposure to long and short artificial "nights." Individuals who live in modern urban environments differ in the degree to which, or even whether, the intrinsic duration of melatonin secretion (the duration measured in constant dim light) responds to seasonal changes in the length of the solar night. Changes in the intrinsic duration of melatonin secretion that are induced by changes in the scotoperiod are highly correlated with changes in the intrinsic timing of the morning offset of secretion and are only weakly correlated with changes in the intrinsic timing of evening onset of secretion. This finding suggests that differences in the way in which individuals are exposed to, or process, morning light may explain differences in their responsiveness to changes in duration of natural and experimental scotoperiods. Although the human RHP axis clearly is capable of detecting changes in the length of the night and in producing the melatonin message that other animals use to trigger seasonal changes in their behavior, it is not yet known whether or how the human reproductive system or other systems respond to this message. PMID- 9406026 TI - Melatonin receptors: molecular biology of a new family of G protein-coupled receptors. AB - A family of high-affinity, G protein-coupled receptors for the pineal hormone melatonin has been cloned from vertebrates. These recombinant receptors exhibits similar affinity and pharmacological characteristics to each other and to endogenous receptors, as defined with the melatonin agonist 2-[125I]iodomelatonin (125I-Mel). Two mammalian melatonin receptor subtypes have been identified by molecular cloning studies. The mammalian Mel1a melatonin receptor is expressed in most sites containing 125I-Mel binding. This includes the hypothalamic suprachiasmatic nucleus and hypophyseal pars tuberalis, presumed sites of the circadian and some of the reproductive actions of melatonin, respectively. The mammalian Mel1b melatonin receptor is expressed in retina and brain and may mediate the reported effects of melatonin on retinal physiology in some mammals. A third receptor subtype, the Mel1c melatonin receptor, has been cloned from zebra fish, Xenopus, and chickens but not from mammals. Molecular cloning of a melatonin receptor family now makes possible gene targeting to precisely determine the physiological role(s) of each receptor subtype. PMID- 9406027 TI - Commentary: what does melatonin do and how does it do it? PMID- 9406028 TI - Photic regulation of melatonin in humans: ocular and neural signal transduction. AB - Light is a potent stimulus for regulating the pineal gland's production of melatonin and the broader circadian system in humans. It initially was thought that only very bright photic stimuli (> or = 2500 lux) could suppress nocturnal melatonin secretion and induce other circadian responses. It is now known that markedly lower illuminances (< or = 200 lux) can acutely suppress melatonin or entrain and phase shift melatonin rhythms when exposure conditions are optimized. The elements for physical/biological stimulus processing that regulate photic influences on melatonin secretion include the physics of the light source, gaze behavior relative to the light source, and the transduction of light energy through the pupil and ocular media. Elements for sensory/neural signal processing become involved as photons are absorbed by retinal photopigments and neural signals are generated in the retinohypothalamic tract. Aspects of this physiology include the ability of the circadian system to integrate photic stimuli spatially and temporally as well as the wavelength sensitivity of the operative photoreceptors. Acute, light-induced suppression of melatonin is proving to be a powerful tool for clarifying how these elements of ocular and neural physiology influence the interaction between light and the secretion of melatonin from the human pineal gland. PMID- 9406029 TI - Photic entrainment of the mammalian rhythm in melatonin production. AB - This review summarizes studies on the photic entrainment of the circadian rhythm in the rat pineal melatonin production, namely of the rhythm in N acetyltransferase (NAT) activity, and compares the NAT rhythm resetting with preliminary results on the resetting of an intrinsic rhythmicity in the suprachiasmatic nucleus (SCN) of the hypothalamus, namely with the entrainment of the rhythm in the light-induced c-fos gene expression. Phase delaying of the NAT rhythm after various light stimuli proceeds within 1 day with almost no transients, whereas during phase advancing of the rhythm only the morning NAT decline is phase advanced within 1 day and the evening rise phase shifts through transients. A light stimulus encompassing the middle of the night may phase delay the evening NAT rise, phase advance the morning decline, compress the rhythm waveform, and eventually lower its amplitude. Similarly, a long photoperiod compresses the NAT rhythm waveform. The magnitude of phase shifts of the NAT rhythm, as well as their direction, depends on a previous photoperiod. Phase shifts of the evening rise in c-fos gene photoinduction in the SCN and of the morning decline are similar to those of the pineal NAT rhythm after all light stimuli studied so far. The data indicate that the resetting of the rhythm in melatonin production in the rat pineal gland reflects changes in the SCN functional state and suggest that the underlying SCN pacemaking system is complex. PMID- 9406030 TI - Resetting the melatonin rhythm with light in humans. AB - The endogenous circadian rhythm of melatonin in humans provides information regarding the resetting response of the human circadian timing system to changes in the light-dark (LD) cycle. Alterations in the LD cycle have both acute and chronic effects on the observed melatonin rhythm. Investigations to date have firmly established that the melatonin rhythm can be reentrained following an inversion of the LD cycle. Exposure to bright light and darkness given over a series of days can rapidly induce large-magnitude phase shifts of the melatonin rhythm. Even single pulses of bright light can shift the timing of the melatonin rhythm. Recent data have demonstrated that lower light intensities than originally believed are capable of resetting the melatonin rhythm and that stimulation of photopically sensitive photoreceptors (i.e., cones) is sufficient to reset the endogenous circadian melatonin rhythm. In addition to phase resetting, exposure to light of critical timing, strength, and duration can attenuate the amplitude of the endogenous circadian rhythm of melatonin. Measurement of melatonin throughout resetting trials provides a dynamic view of the resetting response of the human circadian pacemaker to light. Future studies of the melatonin rhythm in humans may further characterize the resetting response of the human circadian timing system to light. PMID- 9406031 TI - Acute and delayed effects of exercise on human melatonin secretion. AB - Accumulating evidence suggests that exercise may have both rapid and delayed effects on human melatonin secretion. Indeed, exercise may acutely (i.e., within minutes) alter melatonin levels and result in a shift of the onset of nocturnal melatonin 12 to 24 h later. The presence and nature of both acute and delayed effects appear to be dependent on the timing of exercise. The presence of a detectable acute effect also depends on the duration, intensity, and type of exercise. Late evening exercise during the rising phase of melatonin secretion may blunt melatonin levels. High-intensity exercise during the nighttime period, when melatonin levels already are elevated, consistently results in a further (nearly 50%) elevation of melatonin levels. No effect of low-intensity exercise performed at the same circadian phase could be detected. Irrespective of intensity, exercise near the offset of melatonin secretion or during the daytime has no consistent acute effect on melatonin secretion. Nighttime exercise, whether of moderate or high intensity, results in phase delays of the melatonin onset on the next evening. In support of the concept that a shift of the melatonin onset on the day after nighttime exercise represents a shift of intrinsic circadian timing is the observation that similar phase shifts (in both direction and magnitude) may be observed simultaneously for the onset of the circadian elevation of thyrotropin secretion. The observation of exercise-induced phase shifts of the onset of melatonin secretion is, therefore, interpreted as evidence that, in humans as in rodents, increased physical activity during the habitual rest period is capable of altering circadian clock function. PMID- 9406032 TI - Commentary: effects of photic and nonphotic stimuli on melatonin secretion. PMID- 9406033 TI - Circadian entrainment and phase shifting in mammals with melatonin. AB - Circadian effects of exogenous melatonin, whereby daily administration induces entrainment or phase shifts, have been demonstrated in both nocturnal and diurnal mammals. In Long-Evans rats, as used in early studies, effects occur reliably when melatonin is administered late in the subjective day. A second period of sensitivity to melatonin, late in the subjective night, is evident in certain strains of mice and the diurnal Funambulus pennanti. This late night to early morning sensitive phase previously had been identified in human subjects. Different circadian responses to melatonin also may occur between rat strains. Circadian effects of melatonin in hamsters are diverse and vary with strain, developmental age, and method of administration. Characteristics of melatonin binding sites within the suprachiasmatic nuclei vary both between and within species, as do profiles of endogenous melatonin rhythms. These differences may explain the variations in circadian responses to melatonin. PMID- 9406034 TI - Exogenous melatonin's phase-shifting effects on the endogenous melatonin profile in sighted humans: a brief review and critique of the literature. AB - Melatonin's phase-shifting effects in humans are thought by some investigators to be subtle, particularly in comparison to those achieved with appropriately timed bright light exposure. The initial study in sighted people was only intermittently successful in phase advancing the endogenous melatonin profile. The study of free-running blind people showed statistically significant phase advances the day after melatonin administration. When holding the light-dark cycle constant, consistent phase advances the day after melatonin administration in sighted people were first shown in the course of describing the melatonin phase response curve (PRC), which also provided the first evidence that melatonin could cause phase delays. More recent studies have replicated the PRC and shown that phase shifts can occur in response to physiological doses within 1 day. This article reviews this literature and attempts to reconcile some of the results from differing studies. If the timing of melatonin administration is optimized according to the melatonin PRC, then consistent phase advances and delays can be achieved. If a reliable and sensitive circadian phase marker (e.g., the highly resolved dim light melatonin onset) is used, then phase shifts can be demonstrated consistently--even a small shift the day after a single physiological dose. The present authors predict that in the near future, melatonin administration will become as useful as bright light exposure in the treatment of circadian phase disorders. PMID- 9406035 TI - Melatonin as a chronobiotic: treatment of circadian desynchrony in night workers and the blind. AB - Although the causes are different, totally blind people (without light perception) and night shift workers have in common recurrent bouts of insomnia and wake-time sleepiness that occur when their preferred (or mandated) sleep and wake times are out of synchrony with their endogenous circadian rhythms. In this article, the patterns of circadian desynchrony in these two populations are briefly reviewed with special emphasis on longitudinal studies in individual subjects that used the timing of melatonin secretion as a circadian marker. In totally blind people, the most commonly observed pattern is a free-running rhythm with a stable non-24-h circadian period (24.2-24.5 h), although some subjectively blind people are normally entrained, perhaps by residually intact retinoypothalamic photic pathways. Experiments at the cellular and behavioral levels have shown that melatonin can produce time dependent circadian phase shifts. With this in mind, melatonin has been administered to blind people in an attempt to entrain abnormal circadian rhythms, and substantial phase shifts have been accomplished; however, it remains to be demonstrated unequivocally that normal long-term entrainment can be produced. In untreated night shift workers, the degree and direction of phase shifting in response to an inverted sleep-wake schedule appears to be quite variable. When given at the optimal circadian time, melatonin treatment appears to facilitate phase shifting in the desired direction. Melatonin given prior to a night worker's daytime sleep also may attenuate interference from the circadian alerting process. Because melatonin has both phase-shifting and sleep-promoting actions, night shift workers, who number in the millions, may be the most likely group to benefit from treatment. PMID- 9406036 TI - Efficacy of melatonin treatment in jet lag, shift work, and blindness. AB - Melatonin has chronobiotic properties in humans. It is able to phase shift strongly endogenous rhythms, such as core temperature and its own endogenous rhythm, together with the sleep-wake cycle. Its ability to synchronize free running rhythms has not been fully investigated in humans. There is evidence for synchronization of the sleep-wake cycle, but the available data suggest that it is less effective with regard to endogenous melatonin and core temperature rhythms. When suitably timed, most studies indicate that fast release preparations are able to hasten adaptation to phase shift in both field and simulation studies of jet lag and shift work. Both subjective and objective measures support this statement. However, not all studies have been successful. Careful evaluation of the effects on work-related performance is required. When used to alleviate the non-24-h sleep-wake disorder in blind subjects, again most studies report a successful outcome using behavioral measures, albeit in a small number of individuals. The present data suggest, however, that although sleep wake can be stabilized to 24 h, entrainment of other rhythms is exceptionally rare. PMID- 9406037 TI - Commentary: evidence for melatonin as a circadian phase-shifting agent. PMID- 9406038 TI - Melatonin and the circadian regulation of sleep initiation, consolidation, structure, and the sleep EEG. AB - The endogenous circadian rhythm of melatonin, driven by the suprachiasmatic nucleus, exhibits a close association with the endogenous circadian component of the sleep propensity rhythm and the endogenous circadian component of the variation in electroencephalogram (EEG) oscillations such as sleep spindles and slow waves. This association is maintained even when the sleep-wake cycle is desynchronized from the endogenous circadian rhythm of melatonin. Administration of melatonin during the day increases daytime sleep propensity as indexed by both the latency to sleep onset and sleep consolidation. The EEG during daytime sleep after melatonin administration exhibits characteristics reminiscent of the nocturnal sleep EEG, that is, increased sleep spindle activity and reduced slow wave sleep and slow-wave activity, as detected by quantitative EEG analysis. Administration of higher doses of melatonin (5 mg or more) prior to nocturnal sleep results in an increase in rapid eye movement (REM) sleep. These data demonstrate that melatonin exerts effects on the main characteristics of human sleep, that is, latency to sleep onset, sleep consolidation, slow waves, sleep spindles, and REM sleep. There is a need for further studies using physiological doses and delivery systems that generate physiological plasma melatonin profiles to firmly establish the role of the endogenous circadian rhythm of melatonin in the circadian regulation of sleep. PMID- 9406039 TI - The acute soporific action of daytime melatonin administration: effects on the EEG during wakefulness and subjective alertness. AB - Melatonin has been reported to have soporific effects; following daytime administration, it induces sleepiness and reduces sleep onset latency. However, subjective sleepiness is masked by a variety of stimuli and behaviors; thus, it is important to be able to delineate objective psychophysiological sequelae of melatonin administration. Alertness decrements during wakefulness are correlated with augmented theta/alpha power in the waking electroencephalogram (EEG). This has been validated in a constant routine protocol. In a variety of experiments with melatonin administration (5 mg), the authors have shown that the EEG changes can be measured immediately, before any subjective soporific effects are recognized. These increases in theta/alpha power occur when melatonin is administered during the day (1300 or 1800 h) but are less visible when near the endogenous melatonin rise in the evening (2040 h). Importantly, both subjective and objective measures of sleepiness are suppressed when subjects change posture from supine to standing. PMID- 9406040 TI - Efficacy of melatonin as a sleep-promoting agent. AB - Numerous studies have demonstrated sleep-promoting effects of melatonin treatment in humans, as evidenced by subjects' self-reports, polysomnographic recordings, and continuous actigraphic registration of motor activity. The sleep-promoting effects of either physiological or pharmacological doses of melatonin typically are observed within 1 h following treatment regardless of the time of melatonin administration. This fact indicates the acute nature of this effect of melatonin on sleep, independent of any effect of the melatonin treatment on circadian organization. This article considers a dose dependency of melatonin effects on sleep, interindividual variability, and age-related differences in circulating melatonin levels produced in response to a given dose of the hormone. Possible side effects of melatonin treatment, and the use of an animal model to serve as a guide in the development of therapeutic applications, also are considered. PMID- 9406041 TI - Efficacy of melatonin as a hypnotic agent. AB - The seemingly contradictory literature on the use of melatonin in insomnia is best understood by considering issues of time of administration (day or night), dose range (physiological or pharmacological), subject selection (normals or insomniacs), choice of dependent variable (self-report or electroencephalogram), and comprehensiveness of the reported variables (sleep onset or sleep maintenance). Available data on nighttime administration to normals and insomniacs are reviewed. It is concluded that there is not yet a convincing body of evidence, using generally accepted measures, that melatonin administration improves sleep in insomniacs with noncircadian sleep disturbance. PMID- 9406042 TI - Melatonin: role in gating nocturnal rise in sleep propensity. AB - The present article reviews the evidence that melatonin possesses sleep-inducing effects and that it gates the increase in nocturnal sleepiness. It is shown that, without exception, all the studies that have investigated daytime administrations of melatonin reported increased sleepiness, even at doses that do not increase plasma levels of melatonin beyond its physiological levels. By contrast, nighttime increase in sleepiness was achieved only after administration of high doses. Based on these findings and on the precise coupling between the endogenous nocturnal increase in melatonin secretion and the opening of the sleep gate, it is suggested that melatonin participates in the regulation of the sleep-wake cycle by inhibiting the central nervous system wakefulness generating system. This inhibition allows a smooth transition from wakefulness to sleep. Clinical findings on decreased levels of nocturnal melatonin in chronic insomniacs, and on the efficacy of exogenous melatonin in improving sleep in melatonin-deficient insomniacs, are congruent with this hypothesis. PMID- 9406043 TI - Commentary: is melatonin administration an effective hypnotic? PMID- 9406044 TI - Safety of melatonin in long-term use (?) AB - There are no published long-term safety data on the use of melatonin for whatever purpose, assuming long term to mean more than 6 months of daily medication. In the light of its physiological role in animals, the potential deleterious effects include inhibition of reproductive function, delayed timing of puberty, and influence (when taken during pregnancy and lactation) on the circadian status of the fetus and neonate and on future development. Its interactions with other medications are virtually unexplored. For most positive effects published, there also exist negative reports. There are insufficient data on its use in organic or psychiatric disease for any evaluations to be made. There are insufficient data on dose, formulation, and consequent relationships of individual pharmacokinetics and pharmacodynamics for recommendations at present. However, in normal healthy adults over 18 years old, not pregnant or lactating, with no personal or family histories of psychiatric disorder, and unmedicated except for oral contraceptives and minor analgesics (if necessary), the only significant short-term side effect in the author's experience has been sleepiness following oral ingestion of synthetic melatonin (5 mg or less, oral fast release), licensed for human experimental use and for prescription on a named-patient basis. PMID- 9406045 TI - Reproductive safety of melatonin: a "wonder drug" to wonder about. AB - By some accounts, melatonin is the wonder drug of the 1990s. This previously obscure hormone came to the public's full attention as the result of a series of popular books claiming therapeutic benefits of melatonin ingestion. Some of these claims deserve serious consideration and investigation, whereas others appear unfounded. Without waiting for the outcome of the ongoing scientific debate, however, melatonin set astounding sales records. The hormone is now ingested on a daily basis by many thousands of people. There is little information on the potential adverse effects of melatonin ingestion in humans. Melatonin, its analogs, and its metabolites are not mutagenic, and melatonin possesses remarkably low acute toxicity in animals and humans. It is more difficult to exclude toxic effects of long-term melatonin treatment. The fact that melatonin is normally secreted each night does not ensure that exogenous melatonin, taken at other times and/or in supraphysiological doses, will not have adverse effects. Despite the well-recognized role of melatonin in the regulation of reproduction in photoperiodic species, it seems unlikely that chronic ingestion of moderate melatonin doses will have a profound impact on reproductive function in humans. Evidence that melatonin modulates steroid hormone action in some steroid responsive tissues suggests that these tissues should be carefully examined when attempting to assess whether melatonin has chronic toxicity in humans. In the absence of sufficient information regarding the longterm safety of exogenous melatonin, the conservative course of action is to restrict melatonin use to those therapeutic applications in which a significant benefit is expected. The decision to ingest melatonin should be preceded by careful consideration of the expected benefits as well as the potential costs of treatment, with recognition of the fact that there has been exaggeration of the benefits and little attention paid to the potential costs in most discussions of this issue to date. PMID- 9406048 TI - Commentary on the articles by Arendt, Weaver, Mahle et al., and Guardiola Lemaitre. PMID- 9406047 TI - Toxicology of melatonin. AB - Despite the fact that melatonin has been released for public use in the United States by the Food and Drug Administration and is available over the counter nationwide, there currently is a total lack of information on the toxicology of melatonin. In Europe, melatonin has a completely different status in that it is considered a "neurohormone" and cannot be sold over the counter. Even though administration of melatonin in humans, as well as in animals (even at supraphysiological doses), has not shown evidence of toxicological effects (i.e., no deaths), a drug toxicological file still would need to be prepared and approved by the regulatory authorities. Several features that are specific to this neurohormone need to be taken into consideration. Whatever the species concerned, melatonin is secreted during the night; it is the "hormone of darkness." It presents a circadian rhythm and a circannual rhythm (in photoperiodic species). The duration of these secretions could have an impact on the reproductive system, for example, showing the importance of the pharmacodynamics of melatonin. An inappropriate time schedule of melatonin administration could induce supraphysiological concentrations of the neurohormone and a desensitization of melatonin receptors. A long duration of exposure to melatonin also could mimic an "artificial darkness" condition when a circadian rhythm with a basal zero level during the day needs to be conserved for a physiological function. Furthermore, administration of large doses of melatonin could induce high concentrations of melatonin and of different metabolites that could have deleterious effects per se. Numerous books, magazines, and articles have praised melatonin as a "miraculous cure-all" for ailments ranging from sleeplessness, to aging, without any clinical evidence of efficacy (with the exception of its chronobiotic and resynchronizing effect). Very little attention has been paid to the possible side effects of melatonin. Nightmares, hypotension, sleep disorders, abdominal pain, etcetera, have been reported. In fact, analysis of the known pharmacological profile of melatonin and/or of its metabolites, based on scientific preclinical studies, constitutes a basis for prediction of adverse drug reactions or side effects. These include (1) the central nervous system, (2) the cardiovascular system and platelet aggregation, (3) glucose metabolism, (4) immunology, and (5) cancer. The knowledge of the fundamental mechanism of action of melatonin, including molecular biology, also needs to be taken into account for evaluation of possible side effects. Two types of melatonin receptors have been cloned (related to cyclic AMP), and the possibility of intracellular action of melatonin cannot be excluded. Melatonin receptors are present in the periphery and also at the level of the central nervous system, particularly on the suprachiasmatic nucleus that "drives" a circadian rhythm to many other areas on which it projects. Among those, the hypothalamus (which has melatonin receptors) plays a fundamental role in the hormonal homeostasis and modulation control of the organism. Special preclinical and pharmacological studies that take into account all these parameters need to be designed for safety evaluation and risk assessment of this specific neurohormone. PMID- 9406046 TI - Melatonin modulates vascular smooth muscle tone. AB - The molecular cloning of a family of melatonin receptors has created a renewed interest in the diverse actions of the hormone melatonin. The radioligand 2 [125I]iodomelatonin has identified potential sites of action for melatonin throughout the central nervous system and periphery of numerous species. Interestingly, in addition to the suprachiasmatic nucleus (the "biological clock"), 2-[125I]iodomelatonin binding sites have been localized to the rat caudal and cerebral arteries. Furthermore, in vitro, melatonin has been shown to induce a concentration-dependent vasoconstriction of rat caudal and cerebral arteries, and pig and human coronary arteries. The lack of melatonin receptor subtype-selective agonists and antagonists prevents the full pharmacological characterization of these responses. The physiological significance of the in vitro vasoconstrictive properties is far from clear, however; in rats, melatonin has been shown to reduce cerebral blood flow. The widespread use of melatonin warrants appropriately designed studies to probe the role of melatonin and its receptors in the modulation of in vitro vascular tone. PMID- 9406049 TI - Reactivity of recombinant Treponema pallidum (r-Tp) antigens with anti-Tp antibodies in human syphilitic sera evaluated by ELISA. AB - We evaluated the immunoreactivity of recombinant Treponema pallidum (r-Tp) antigens with human sera by indirect enzyme-linked immunoabsorbant assay (ELISA). We expressed antigens with a molecular weight (MW) of 17KDa, 15KDa, 47KDa, and 42KDa, which are believed to be major immunoreactive membrane proteins of Tp cells. The expressed proteins were described by adding the prefix M, S, and G to the corresponding Tp antigens, namely, mature antigens, signal sequence containing antigens, and glutathione s-transferase (GST)-fused antigen in this report. A rather high expression occurred for M47 and S42 proteins in the Escherichia coli system, whereas for M15 and M17 proteins, a poor expression was observed. However, a fairly high expression occurred for G15 and G17. Thus expressed proteins were purified by means of chromatographies to a level of > 95%, and the purified proteins were found to be reactive with TPHA positive serum by Western blotting (WB). An ELISA performed with a serum of 1/1000 dilution using these purified antigens for coating on the solid phase showed that G17 antigen was more effective in detecting syphilis antibodies in human serum than M47, S42, and G15. There was a good consistency between ELISA and TPHA, whereby the cutoff indexes (CI) on ELISA showed a correlation coefficient of 0.7276 in logarithmic TPHA titers. PMID- 9406050 TI - Optimized PCR amplification of influenza A virus RNA using Tth DNA polymerase, incorporating uracil N glycosylase (UNG) in a single tube reaction. AB - An optimized reaction condition for amplification of influenza A virus RNA, by thermus thermophilus (Tth) DNA polymerase-based PCR, incorporating uracil N glycosylase (UNG) and dUTP in the reaction has been determined. dUTP could not be substituted for all dTTP sites when UNG was present in the reaction. The relative concentration of dUTP and dTTP has been optimized for allowing amplification of the target RNA. It has been verified that the amplified product DNA had sufficient dUTP and was digestable by UNG. Using the optimized reaction condition, influenza A virus-specific DNA fragment could be amplified and detected in 15 of 15 culture positive (for influenza A virus) nasopharyngeal specimens. PMID- 9406051 TI - Detection of a common mutation in factor V gene responsible for resistance to activate protein C causing predisposition to thrombosis. AB - Hereditary predisposition to thrombosis due to activated protein C resistance (APCR) has been attributed to a missense mutation in the factor V gene at nucleotide 1691 (G to A), causing replacement of arginine at codon 506 with glutamine. Using an RFLP-PCR assay to detect this mutation, we measured a prevalence of 3.3% in healthy Caucasians and 1.25% in healthy African-Americans. In addition, we evaluated a total of 90 consecutive specimens submitted to the coagulation laboratory at the Medical College of Virginia for the presence of this mutation. We compared our results for 78 of these specimens with the values measured by a modified partial thromboplastin assay, the COATEST. Twelve of the 90 samples could not be tested using the COATEST because the patients were undergoing anticoagulant therapy. One of the latter 12 specimens was positive by the RFLP-PCR test. Using the genetic test as the definitive assay and the cutoff value established for distinguishing between normal and abnormal results by the COATEST, the COATEST had a sensitivity of 50% and specificity of 93% for the detection of factor V mutation. Analysis of the 90 samples stratified by ethnic groups revealed a frequency of mutation of 13.3% for Caucasians and 6.88% for African-Americans, although with the present sample size, the difference was not statistically significant. Although the COATEST is technically simpler to perform than the genetic test for diagnosing the presence of the factor V mutation, its use for this purpose is limited due to low sensitivity. Thus where this disorder is clinically suspected, submission of the specimen directly for genetic testing by RFLP-PCR or equivalent assay should be considered. PMID- 9406052 TI - Measurement of plasma ibuprofen by gas chromatography-mass spectrometry. AB - A gas-chromatography-mass spectrometry (GC-MS) method for the determination of plasma ibuprofen was developed. Plasma samples from cystic fibrosis (CF) patients receiving high-dose ibuprofen therapy were analyzed by GC-MS and the result compared to analysis by high-performance liquid chromatography (reference method). Analysis of ibuprofen was sensitive to at least 5 mg/L, and the method was linear to 200 mg/L. Within-run variations of plasma samples were 4.6% (131.7 +/- 6.0 mg/L) and 5.4% (44.4 +/- 2.4 mg/L), respectively. The between-run variation was 9.3% (45.4 +/- 4.2 mg/L) and 7.4% (88.0 +/- 6.5 mg/L). This method is suited for routine clinical use for the monitoring of plasma ibuprofen levels in treatment of CF. It may be particularly applicable in pediatric laboratories, which are likely to possess GC-MS capability. PMID- 9406053 TI - Fast isolation of RNA to detect expression of tumor markers. AB - The expression status of several tumor-related proteins is of great interest in clinical examination and research. As a completion to conventional antibody staining, RT-PCR is often used today. Reliable isolation of RNA from a low number of cells is very often a critical stage of such an examination. We demonstrate here a simple and fast method to isolate RNA from only 10,000 cells and applied it to the detection of CEA, c-ERB-B2, and mdr-1 as often studied models for tumor markers. PMID- 9406054 TI - Analysis of HIV seropositive thalassemic children for antibodies specific to Aspergillus fumigatus by luminescent immunoassay. AB - The applicability of luminescent immunoassay (LIA) in serodiagnosis of fungal infections in multitransfused (MT) thalassemic children seropositive for human immunodeficiency virus (HIV) was investigated. Thirty-one sera samples from HIV infected pediatric patients with thalassemia receiving repeated blood transfusions were analysed for the presence of antibodies specific to Aspergillus fumigatus by LIA. The LIA was standardized using well defined antigens of A. fumigatus. Ten out of 31 (32.2%) of the MT-HIV positive patients were found to have anti-Aspergillus antibodies in their sera by LIA. The ELISA could detect A. fumigatus specific antibodies in 25.8% (8 out of 31) of the patients. Thus, 20% more number of patients turned to be positive for aspergillosis by LIA as compared to ELISA. The difference was found to be statistically significant (p < 0.005). Of the MT-HIV negative patients only 1 out of 33 (3%) showed A. fumigatus specific antibodies by LIA and ELISA both. In age and sex matched control group (n = 25) none of the patients was found to be positive for antibodies to A. fumigatus. LIA was found to have better discriminatory value indicating, thereby, its utility in diagnosis of aspergillosis in compromised patients. PMID- 9406055 TI - Application of immunomagnetic beads in combination with RT-PCR for the detection of circulating prostate cancer cells. AB - Recently published protocols using Reverse Transcriptase Polymerase Chain reaction (RT-PCR) for prostate specific antigen (PSA) provide a sensitive means for detecting circulating prostate cancer cells. Attempts to use these assays for staging of prostate cancer have produced conflicting results. As a first step towards rectifying these discrepancies, a modified immunobead-RT-PCR assay capable of detecting as few as 10 prostate cancer cells in 8cc of blood was developed. This 10 fold increase in sensitivity was achieved in part by introducing two target cell enrichment steps. As a model system to assess sensitivity of the modified assay, template RNA was extracted from PSA positive human carcinoma cells suspended in human blood and isolated with immunomagnetic beads following incubation with an epithelium specific antibody. After 45 cycles of PCR, product from as few as 10 target cells could be readily detected when displayed on a 2% agarose gel stained with SYBR Green fluorescent dye. The identity of amplified DNA fragments was confirmed by Southern blot hybridization. When applied to blood samples from patients with proven metastatic disease, the immuno-bead RT-PCR assay was successful in detecting circulating PSA positive epithelial cells, suggesting this assay may be useful for assessment of disease progression or recurrence. PMID- 9406056 TI - Erythrocyte adenylate kinase isoenzyme as a marker for hemolysis. AB - The presence in serum of adenylate kinase isoenzyme originating from erythrocyte can be useful as a marker for detecting hemolysis. We have presented preliminary evidence for identifying hemolytic anemia patients earlier by determining erythrocyte AK isoenzyme activity in serum (or plasma) rather than using measurement of plasma hemoglobin concentration. This test being quite specific for hemolysis should find use as a quick method for estimating the extent of in vivo hemolysis in hemolytic patients earlier than heretofore possible. PMID- 9406058 TI - Automated measurement of a constitutive isotype of serum amyloid A/SAA4 and comparison with other apolipoproteins. AB - A constitutive isotype of serum amyloid A (SAA4) is present mostly in high density lipoprotein (HDL) and a little in other lipoproteins. In this study, we developed an automated method for measuring SAA4 concentration in serum by kinetic nephelometry of anti-SAA4 antibody-coated latex agglutination. Rabbit antibodies generated by immunization of recombinant SAA4 were found to have no apparent reactivity with acute phase SAA. The values determined by this method and by the previous enzyme immunoassay showed good agreement (r = 0.862). Serum SAA4 values of 26 healthy adults ranged from 37-109 mg/L (mean; 62 mg/L). Their SAA4 concentrations were not significantly related with those of apolipoprotein A I, A-II, B, C-II, C-III or E. Also, SAA4 did not correlate with cholesterol in preparation after removal of very low density lipoprotein and low density lipoprotein. These suggest the unique behavior of SAA4 in lipoprotein metabolism, while what contributes to variation of SAA4 levels in serum, especially in HDL, remains to be clarified. PMID- 9406057 TI - Simultaneous quantitation of specific IgE against 20 purified allergens in allergic patients sera by checkerboard immunoblotting (CBIB). AB - A simple technique, checkerboard immunoblotting (CBIB), is described for simultaneous quantitation of specific IgE antibodies against several allergens in human sera. Using as little as 50 microliters of each of the 20 sera examined against 20 different allergens, it was possible in a single run to achieve 400 tests. To guarantee high specificity and sensitivity of the assay, this new application of CBIB employs purified allergens, cyanogen bromide-activated nitrocellulose membrane, and Phosphorimager technology. Results are expressed both qualitatively (five classes) and quantitatively in kilo units per liter equilibrated against the World Health Organization (WHO) standard for IgE. There was excellent agreement between the results of CBIB and the results of Pharmacia Cap System, an alternative method widely used for measuring serum-specific IgE. The CBIB method certainly could be useful in any laboratory interested in allergy clinical research for easy screening and relative quantitation of allergen specific human IgE. PMID- 9406059 TI - Two color analysis of HLA-B27 antigen by flow cytometer--a comparative study by conventional microlymphocytotoxicity, DNA genotyping polymerase chain reaction and flow cytometric measurement. AB - For evaluation of specificity and sensitivity of flowcytometric determination of HLA-B27 antigen, we determined the HLA-B27 on lymphocytes using HLA-B27 monoclonal antibody by flow cytometer. Data were compared to those by conventional Terasaki microlymphocytoxicity test and DNA genotyping Polymerase Chain Reaction (PCR) method. One hundred and ninety four patients with various forms of arthritis were included in this study. Forty one of them were HLA-B27 positive, confirmed by three methods concomitantly with complete accordance. None of serological B27 negative, B7 CREG positive cells were found to be flowcytometric fluorescence positive. Furthermore, there was no significant difference of B27 intensity between different B27 DNA subtypes, nor was there any difference between primary ankylosing spondylitis (AS) and other secondary spondylitis patients as measured by mean channel of fluorescence. It is suggested that flowcytometric measurement of HLA-B27 antigen is a rapid and reliable method for HLA-B27 determination. PMID- 9406060 TI - In situ hybridization for Helicobacter pylori in gastric mucosal biopsy specimens: quantitative evaluation of test performance in comparison with the CLOtest and thiazine stain. AB - Numerous detection methods for Helicobacter Pylori (H. pylori) have been developed with varying degrees of purported diagnostic utility. We have developed a rapid nonradioactive in situ hybridization (ISH) method for H. pylori detection in paraffin-embedded tissue and assessed its relative diagnostic performance by receiver operator characteristics (ROC) in comparison to the thiazine stain and CLOtest. Forty-five patients undergoing endoscopy had antral biopsies and concomitant CLOtest performed. ISH for H. pylori was done using a 22-base, biotin labeled oligonucleotide probe complementary to a portion of H. pylori 16s rRNA with the following sequence: 5'-GGACATAGGCTGATCTTAGC-3'. ISH using this probe was specific for H. pylori with no crossreactivity with other bacterial or fungal organisms. Receiver operator characteristic analysis was used to assess the diagnostic performance of ISH and thiazine techniques. ISH and thiazine stains were done on serial sections, reviewed independently, and scored on a graded scale from 1-5 based upon the degree of assurance of H. pylori presence. Diagnostic performance was assessed in "expert" and "nonexpert" pathologist groups with the CLOtest serving as the invariant test for relative test comparison. The ISH test performed slightly better (ROC area 0.9) than the thiazine (ROC area 0.8) in the nonexpert population, but equally well in the "expert" group (ROC area 0.95, 0.95). ISH followed by routine hematoxylin and eosin staining showed detailed mucosal histology with a dramatic visualization of H. pylori along the surface of the foveolar cells with no evidence of lamina propria invasion. In summary, ISH for H. pylori is an excellent test that is specific, easily read, and allows concomitant detailed histologic mucosal examination. PMID- 9406061 TI - A sandwich transfer enzyme immunoassay for salmon calcitonin: determination of the bioavailability of intranasal salmon calcitonin in human. AB - A sandwich transfer enzyme immunoassay for salmon calcitonin (SCT) and its usability for the pharmacokinetic study are described. The assay procedure consisted of the reaction of SCT with 2,4-dinitrophenyl biotinyl anti-SCT IgG and anti-SCT Fab'-beta-galactosidase conjugate, trapping onto (anti-2,4-dinitrophenyl bovine serum albumin) IgG-coated polystyrene balls, eluting with epsilon N-2,4 dinitrophenyl-L-lysine and transferring to streptavidin-coated polystyrene balls and fluorometric detection of beta-D-galactosidase activity. The practical detection limit of SCT was 0.05 pg (15 amol)/50 microliters of sample and 1 pg/ml as the concentration. The application of this method has enabled us to directly estimate the bioavailability of SCT dosed intranasally at the therapeutic level (160 IU, 31 micrograms) for its anti-osteoporotic effect as compared to an intramuscular dose (10 IU, 1.9 micrograms). The pharmacokinetic parameters of the intranasal SCT (n = 6) thus estimated were as follows: the area under the blood concentration-time curve (AUC) 9400 +/- 5400 (SD) pg.h/ml, and the mean residence time (MRT) = 42 +/- 14 (SD) min, when the AUC for the intramuscular SCT (n = 3) = 5600 +/- 2000 (SD) pg.h/ml and the MRT = 39 +/- 19 (SD) min. PMID- 9406062 TI - Comparison and variation of different methodologies for the detection of autoantibodies to nuclear antigens (ANA). AB - Interest in the assessment of autoantibody specificity stems from the need for an autoantibody marker capable of predicting clinical events in autoimmune disorders. However, the multiplicity of epitopes present on autoantigenic particles, the quantitative and qualitative heterogeneity of autoantibodies, as well as the nature of the tests, mean that each of the assays used in their determination have different characteristics. The aim of this study was to compare the specificities of different ANAs using four commercial assays. The routine method used for the detection of ANA is indirect immunofluorescence on Hep-2 cells. The assays used were: counterimmunoelectrophoresis (CIE), enzyme linked immunosorbent assay (ELISA), and two immunoblotting assays. Kappa statistic was applied to evaluate the consistency between tests. Kappa index is a measure of agreement between categorical data. Kappa has a maximum of 1.00 when the agreement is perfect, a value of zero indicates no agreement better than chance, and negative values show worse than chance agreement. For SS-B antibodies, there was a good concordance between all four methods used (Kappa 0.66-0.74). For anti RNP antibodies, the results for CIE/ELISA (Kappa 0.60) were consistent as were the two immunoblot methods (Kappa 0.69). For anti Scl-70 (topoisomerase I) antibody, results from the ELISA and CIE methods were totally consistent (Kappa 1.00). In spite of the high prevalence of anti SS-A/Ro antibodies, the agreement between the methods was poor, without statistical significance. Finally, for Sm antibodies, more consistent results were obtained between CIE/ELISA (Kappa 0.51) and between one of the immunoblotting methods and ELISA (Kappa 0.54). In conclusion, CIE concurs mostly with ELISA for anti-RNP, Scl-70, Sm and SS-B antibodies, but with some disagreement for SS-A antibodies. PMID- 9406063 TI - Determination of the marker residue of albendazole in cheese by ion-pair liquid chromatography and fluorescence detection. AB - A liquid chromatographic method is described that allows quantitative determination of the marker residue of albendazole in cheese. Samples were extracted with acetonitrile, and the extracts were defatted with hexane, evaporated to dryness, reconstituted in ethyl acetate, and purified by partitioning with phosphate buffer. Separation of the sulfoxide, sulfone, and 2 aminosulfone metabolites that constitute the marker residue of albendazole was carried out isocratically with a mobile phase containing both positively and negatively charged pairing ions; detection was performed fluorometrically, with excitation and emission wavelengths of 290 and 320 nm, respectively. Overall recoveries ranged from 73.7 to 84.9%. Precision data based on variation within and between days suggested overall values for relative standard deviation of 3.0 to 3.9%. The sensitivity of the method permitted low limits of detection, particularly for the sulfone and 2-aminosulfone metabolites. PMID- 9406064 TI - A modified form of a vitamin B12 compound extracted from whey fermented by Lactobacillus helveticus. AB - The content of vitamin B12 in whey was reduced considerably during lactic acid fermentation, which was analogous to the lactic acid fermentation of milk. This apparent B12 decrease in whey was caused by B12 compounds that were unextractable by conventional extraction with potassium cyanide but that could be extracted by sonication and treatment by proteases such as pepsin and papain. Forms of the extracted B12 compounds were examined by bioautographies with Escherichia coli 215, coupled with cellulose acetate membrane electrophoresis or HPLC, and were identified as adenosylcobalamin, cyanocobalamin, and hydoroxocobalamin. A considerable amount of unidentified B12 was also detected, and this unidentified B12 compound seems to be the principal B12 compound that was unextractable from the cells. PMID- 9406065 TI - Differential cytokine production in clonal macrophage and T-cell lines cultured with bifidobacteria. AB - When used in commercial fermented dairy products, bifidobacteria may enhance immunity by stimulating cytokine secretion by leukocytes. To assess whether interaction between bifidobacteria and leukocytes promote cytokine production, we cultured RAW 264.7 cells (macrophage model) and EL-4.IL-2 thymoma cells (helper T cell model) in the presence of 14 representative strains of heat-killed bifidobacteria. In unstimulated RAW 264.7 cells, all bifidobacteria induced pronounced increases (up to several hundred-fold) in the production of tumor necrosis factor-alpha compared with that of controls. Interleukin-6 production by unstimulated cells also increased significantly, but less than did tumor necrosis factor-alpha. Upon concurrent stimulation of RAW 264.7 cells with lipopolysaccharide, production of tumor necrosis factor-alpha and interleukin-6 were both enhanced between 1.5- to 5.8-fold and 4.7- to 7.9-fold, respectively, when cultured with 10(8) bifidobacteria/ml. In unstimulated EL-4.IL-2 cells, bifidobacteria had no effect on the production of interleukin-2 or interleukin-5. Upon stimulation of EL-4.IL-2 with phorbol-12-myristate-13-acetate, there were variable increases in interleukin-2 secretion (up to 2.4-fold for 10(6) Bifidobacterium Bf-1/ml) and interleukin-5 secretion (up to 4.6-fold for 10(8) B. adolescentis M101-4). The results indicated that, even when variations among strains were considered, direct interaction of most bifidobacteria with macrophages enhanced cytokine production, but the effects on cytokine production by the T-cell model were less marked. Interestingly, the 4 bifidobacteria strains used commercially for diary foods showed the greatest capacity for cytokine stimulation. The in vitro approaches employed here should be useful in future characterization of the effects of bifidobacteria on gastrointestinal and systemic immunity. PMID- 9406066 TI - Influence of yogurt and acidophilus yogurt on serum cholesterol levels in mice. AB - The effects of yogurt and acidophilus yogurt on the weight gain, serum cholesterol, high density lipoprotein cholesterol, low density lipoprotein cholesterol, triglycerides, and the numbers of fecal lactobacilli and coliforms were investigated in mice assigned to three dietary treatments for 56 d: 1) commercial rodent chow and water (control), 2) commercial rodent chow and yogurt made from milk inoculated with a 3% (vol/vol) liquid culture of Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus (yogurt), and 3) commercial rodent chow plus yogurt made from milk inoculated with a 0.01% (wt/vol) freeze-dried culture of Streptococcus thermophilus plus Lactobacillus acidophilus. The weight gains of mice receiving yogurt or acidophilus yogurt were higher than those of the mice in the control group. The mean values for serum cholesterol concentrations and LDL cholesterol concentrations were significantly decreased when acidophilus yogurt was fed on d 28 and 56. High density lipoprotein cholesterol and triglycerides were not affected by yogurt or acidophilus yogurt. The highest number of fecal lactobacilli was found in mice receiving acidophilus yogurt, and the number of fecal coliforms of that group was also lower than in the other two groups. PMID- 9406067 TI - Evaluation of different protocols for prostaglandin synchronization to improve reproductive performance in dairy herds with low estrus detection efficiency. AB - The objective of this study was to evaluate different PGF2 alpha protocols against control protocols for herds with estrus detection efficiencies of 35, 55, and 75% using modeling and simulation: 1) PGF2 alpha treatments based on the presence of a corpus luteum diagnosed by rectal palpation, 2) PGF2 alpha treatments based on the presence of a corpus luteum diagnosed by an on-farm milk progesterone enzyme immunoassay, and 3) PGF2 alpha treatments based on a 14-d fixed treatment schedule without prior screening for ovarian status. After the start of each protocol, estrus detection efficiency was 75% for 7 d after treatment and 35 or 0% for the following week. For the third protocol, an additional modification at estrus detection efficiencies of 85 and 55%, respectively, in the 1st and 2nd wk after treatment was evaluated to establish a protocol for best case assumptions. All protocols improved reproductive performance relative to that of controls with estrus detection efficiencies of 35 and 55%. The mean number of days open was reduced from 124.3 d in the control herd to 95.9, 95.0, and 92.7 for the protocols based on rectal palpation, milk progesterone test, and the fixed treatment schedule, respectively. The protocols based on a fixed treatment schedule were superior to protocols based on rectal palpation and on-farm milk progesterone tests and resulted in better reproductive performance and a higher increase in net return per cow per year. Relative to a control herd with an estrus detection efficiency of 55%, it was cost effective to spend up to $10 per dose of PGF2 alpha, $9 per milk progesterone test, and $6 per rectal palpation. PMID- 9406068 TI - Estimation of economic values of indices for reproductive performance in dairy herds using computer simulation. AB - Relationships between measures for reproductive performance and net revenue were studied using data that had been generated by a stochastic model in a computer simulation program for Ontario dairy herds. These measures included projected calving interval, involuntary culling rate, and adjusted calving interval. Adjusted calving interval was calculated by dividing the projected calving interval for pregnant cows by the fraction of the cows that were not culled for reproductive failure. The regression of adjusted calving interval on net revenue had an R2 of 0.72, which was higher than the R2 of 0.59 obtained by the regression of projected calving interval on net revenue. Hence, the estimation of financial losses from suboptimal reproductive performance was more accurate when adjusted calving interval was used as a measure of this performance than when projected calving interval was used. This difference is because projected calving interval did not consider cows that were culled for reproductive reasons, but those cows contributed to a reduction in profit because of suboptimal reproductive performance. The highest R2 (0.78) was obtained with a model that included projected calving interval and involuntary culling rate. However, use of that model might not be practical because herd operators differ in their ability to distinguish between involuntary and voluntary culling. The mean reduction in net revenue from a 1-d increase in adjusted calving interval was estimated at $4.7 (Canadian) per cow. This economic value was not significantly affected by level of adjusted calving interval, carcass price, feed costs, cost of a replacement heifer, or milk price. PMID- 9406069 TI - Effects of a long daily photoperiod on milk yield and circulating concentrations of insulin-like growth factor-1. AB - Relative to a short daily (24-h) photoperiod, exposure to a long daily photoperiod increases the milk yield of dairy cows. However, the endocrine basis for this phenomenon is unknown. The present study was designed to test the hypothesis that a long daily photoperiod is associated with increased circulating insulin-like growth factor (IGF)-I, a hormone that is galactopoietic in ruminants. Forty lactating cows were exposed to either a natural photoperiod (< or = 13 h of light/d) or to a long daily photoperiod (18 h of light and 6 h of darkness) between January and April 1995. Cows were fed for ad libitum intake a total mixed diet formulated to meet the nutritional demands of lactation. Milk yield and dry matter intake were quantitated each day, and blood samples were collected by coccygeal venipuncture every 14 d. Plasma was harvested and assayed for IGF-I. The long photoperiod increased milk yield relative to the natural photoperiod (36.1 +/ 0.6 vs. 33.9 +/ 0.6 kg/d); the increase became significant after 28 d of treatment and was maintained for the duration of the study. In addition, cows exposed to a long photoperiod had greater circulating concentrations of IGF-I than did cows exposed to the ambient natural photoperiod (60.1 +/ 2.0 vs. 52.6 +/ 2.0 ng/ml). Concentrations of IGF binding protein -2 and -3 in plasma did not differ between treatments. These results support the hypothesis that a long daily photoperiod increases circulating concentrations of IGF-I in lactating cows and reveal a possible endocrine mechanism for the galactopoietic response to a long daily photoperiod. PMID- 9406070 TI - Associations between winter herd management factors and milk protein yield in Ontario dairy herds. AB - A questionnaire was used to investigate associations between farm management practices in winter and milk protein yield. The questionnaire was completed by 500 dairy producers in Ontario, Canada in March 1994. A high positive correlation (0.96) existed between herd mean for protein yield per day per cow and herd mean for milk yield per day per cow. Therefore, most management practices that were associated with the farm mean for protein yield per day per cow were also associated with the farm mean for milk yield per day per cow. After adjustment for farm mean milk and fat yields, farms that housed dry cows as a separate group had higher mean protein yields than did herds that housed dry cows with the milking herd or the bred heifers. A positive association existed between herd mean for days of lactation and protein yield. Because of the correlation between milk and protein yields, these associations were of small magnitude. Thus, the potential benefit of progressive practices in herd management are primarily related to increased yields of milk and milk components in general rather than specifically to an increase in protein yield. PMID- 9406071 TI - Factors associated with bacteriological cure during lactation after therapy for subclinical mastitis caused by Staphylococcus aureus. AB - The objective of this study was to determine the factors that were associated with the cure of subclinical mastitis caused by Staphylococcus aureus treated during lactation. One hundred forty-three quarters that were infected with S. aureus were available from a number of treatment trials. Analysis of these data showed that the most important factors associated with cure were age of the cow, somatic cell count at the time of treatment, presence of the infection in the front quarters, and stage of lactation. Other factors, such as number of infected quarters per cow and sensitivity or resistance of the strain to penicillin, were not significant. Because of the relatively low probability of cure, it is important to know risk factors for cure and, hence, to choose cows for treatment with great care. Using these data, a prediction equation was developed to determine the cure rate of subclinical mastitis caused by S. aureus when treated during lactation. PMID- 9406072 TI - Efficacies of teat germicides containing 0.5% chlorhexidine and 1% iodine during experimental challenge with Staphylococcus aureus and Streptococcus agalactiae. AB - Two germicides, one containing 0.5% chlorhexidine and one containing 1% iodine, were tested for efficacy against new intramammary infections (IMI) caused by Staphylococcus aureus and Streptococcus agalactiae. The model for the experimental challenge in the trials that were designed to test the efficacy of the two postmilking teat dips was recommended by the National Mastitis Council. The 0.5% chlorhexidine product reduced the number of new IMI caused by Staph. aureus by 73.2% and reduced the number of new IMI caused by Strep. agalactiae by 53.9%. The 1% iodine product reduced the number of new IMI caused by Staph. aureus by 75.6% and reduced the number of new IMI caused by Strep. agalactiae by 53.5%. In both trials, the incidence of clinical mastitis was also reduced in dipped quarters compared with control quarters. Use of the two teat dips over the trial periods had no effect on the condition of teat skin or teat ends. PMID- 9406073 TI - Persistence of subclinical intramammary pathogens in goats throughout lactation. AB - The goal of this study was to determine the persistence of caprine intramammary pathogens throughout lactation and to detect the bias in diagnoses when a single milk sample was used. We studied 131 goats throughout 7 mo of lactation. Goats were sampled monthly, and 1834 milk samples were bacteriologically analyzed. One hundred sixty-eight pathogens were isolated: 82.5% were micrococci, 9.5% were Gram-negative bacilli, and 8% were corynebacteria. An intramammary infection (IMI) was considered a true, persistent IMI when the same pathogen was isolated two or more times consecutively from the same half of the udder. One hundred one samples were considered to be truly positive, which produced persistent IMI caused by nine different species (eight Staphylococcus spp. and one Pseudomonas sp.). Statistical relationships were found between staphylococci and true positive diagnosis and between corynebacteria and false-positive diagnosis. No relationship involving Gram-negative bacilli was detected. A single milk sample had a positive predictive value (60%), high sensitivity (96.2%), high specificity (96.1%), and highly negative predictive value (99.8%). PMID- 9406074 TI - Accuracy of methods using somatic cell count and N-acetyl-beta-D-glucosaminidase activity in milk to assess the bacteriological cure of bovine clinical mastitis. AB - This study examined the capability of milk somatic cell count (SCC) and NAGase activity to discriminate between quarters that had been cured versus those that had not been cured at 4 wk after antimicrobial therapy for clinical mastitis. The distribution of microorganisms that were isolated before therapy from 630 quarters with mastitis was as follows: 225 strains of Staphylococcus aureus, 96 strains of coagulase-negative staphylococci, 152 strains of streptococci (Streptococcus dysgalactiae and Streptococcus uberis), and 157 strains of coliform bacteria. Bacteriological cure rates were 35% for mastitis caused by Staph. aureus, 75% for mastitis caused by coagulase-negative staphylococci, 66% for mastitis caused by streptococci, and 72% for mastitis caused by coliforms. Diagnostic accuracy of milk SCC and NAGase and their interquarter ratios for predicting bacteriological status of the control samples was assessed by calculating sensitivity, specificity, and accuracy and by means of receiver operating characteristic analysis. The efficiency of milk SCC and NAGase for predicting bacteriological cure was greatest for cows that had been infected with Staph. aureus. The main problem in detecting coagulase-negative staphylococci was low sensitivity, and the main problem in detecting streptococci and coliforms was low specificity. Receiver operating characteristic analysis is not completely suitable for the detection of mastitis because reference method bacteriology and indirect tests can never fully agree. To assess the recovery of cows from mastitis caused by Staph. aureus, bacteriology should be supplemented with an examination of milk SCC or NAGase activity at threshold values such as those presented here. PMID- 9406075 TI - Concentrations of alpha-Tocopherol after intramammary infusion of Escherichia coli or lipopolysaccharide. AB - Fifteen Holstein cows were used in a trial involving intramammary challenge to determine the effects of acute clinical mastitis on the concentrations of alpha tocopherol in milk and plasma and the concentrations of neutrophils in milk and blood. Cows were assigned to one of three experimental groups challenged by intramammary infusion of lipopolysaccharide, Escherichia coli, or sterile phosphate-buffered saline. All quarters infused with lipopolysaccharide or E. coli were diagnosed with clinical mastitis on d 1 and 2 after challenge. Acute inflammation caused by intramammary infusion of lipopolysaccharide or E. coli resulted in increased concentrations of alpha-tocopherol in milk in challenged quarters but had no effect on concentrations of alpha-tocopherol in plasma. Concentrations of alpha-tocopherol in milk and blood neutrophils did not differ among treatment groups. Concentrations of alpha-tocopherol did not differ between milk and blood neutrophils. Approximately 25% of the alpha-tocopherol in milk from glands with clinical mastitis was associated with neutrophils, and < 10% of the alpha-tocopherol in milk from nonmastitic glands was associated with neutrophils. A shift toward sources of alpha-tocopherol other than synthesized milk fat occurred during acute inflammation in the mammary gland. PMID- 9406076 TI - Reactivity and phenotype of mononuclear leukocytes from nongravid heifers after in vitro exposure to 9,13-di-cis-retinoic acid. AB - The predominant isomer of retinoic acid in the plasma of dairy cows during the periparturient period is 9,13-di-cis-retinoic acid. Because retinoic acids influence the activity of cells in a variety of tissues, including the immune system, the potential for this isomer to modulate the bovine immune system during the periparturient period must be considered. The present study examined the in vitro effects of 9,13-di-cis-retinoic acid on the reactivity and phenotype of blood mononuclear leukocytes from nongravid Holstein heifers that were sensitized to antigens and that had naturally low plasma concentrations of 9,13-di-cis retinoic acid. In this system, 9,13-di-cis-retinoic acid, approximating the highest plasma concentrations occurring in vivo during the periparturient period, had no effect on DNA synthesis, secretion of interleukin-2 or interferon-gamma, or secretion of immunoglobulin by unstimulated cultures or cultures stimulated by mitogen (pokeweed mitogen) or antigen (ovalbumin). The composition of unstimulated and stimulated mononuclear leukocyte populations, based on percentages of specific cell types, was unaffected by 9,13-di-cis-retinoic acid at the physiologic concentration of 10(-8) M. 9,13-di-cis-Retinoic acid did not affect the actual number of cells in unstimulated cultures and cultures stimulated by antigen but did cause a moderate reduction in the number of cells, primarily CD4+ lymphocytes, in cultures stimulated by mitogen. Overall, these results suggest that the elevated concentration of 9,13-di-cis-retinoic acid in maternal plasma may have a negligible effect on the reactivity and phenotype of cells constituting the circulating mononuclear leukocyte population. PMID- 9406077 TI - Application of a U-13C-labeled amino acid tracer in lactating dairy goats for simultaneous measurements of the flux of amino acids in plasma and the partition of amino acids to the mammary gland. AB - A preliminary study was conducted using lactating British Saanen goats (n = 5) at 109 to 213 d in milk that yielded 1.67 to 3.68 kg of milk/d to examine the application of a U-13C-labeled amino acid (AA) mixture obtained from hydrolyzed algal proteins as a tracer for measuring plasma flux (n = 5) and partition to the mammary gland (n = 3; arteriovenous difference) of 13 AA simultaneously. Except for Ile and Ser, there was incomplete (6 to 54%) equilibration of the tracer with AA from packed blood cells (> 90% erythrocytes) during the 6-h infusions. This result agreed with the large ratio of packed cells to gradients for plasma AA concentration that was also observed. However, net mass and isotope removals by the mammary gland were predominantly from plasma, indicating that the erythrocytes did not participate in kinetic exchanges. Plasma AA fluxes (millimoles per kilogram of metabolizable protein intake per kilogram of body weight 0.75) differed among goats that consumed different protein sources; however, overall rates were lowest for Met (5 to 14) and His (8 to 17) and highest for Leu (48 to 70) and Ala (53 to 88). On average, 25% of plasma flux was partitioned to the mammary gland. Less than 20% of His, Ser, Phe, and Ala were directed to the mammary gland; 20 to 30% of Arg, Thr, Tyr, and Leu were directed to the mammary gland; and 30 to 40% of Pro, Ile, Lys, and Val were directed to the mammary gland. The unidirectional AA flux in the mammary gland (AA apparently available for protein syntheses, oxidation, and metabolite formation) did not match the pattern that is required for casein synthesis, suggesting differences in the metabolic requirements of AA for nonmilk protein synthesis. PMID- 9406078 TI - Effects of a duodenal glucose infusion on the relationship between plasma concentrations of glucose and insulin in dairy cows. AB - The effects of duodenal infusion of glucose on the relationship between plasma concentrations of glucose and insulin and on milk composition were investigated in a crossover design. Eight dairy cows were continually infused with water (control) or glucose (1.5 kg/d). Cows received diets consisting of dehydrated whole-plant maize in restricted amounts to equalize the energy supply between treatments. Basal (before meal) plasma concentrations of glucose and insulin were increased, but concentrations of nonesterified fatty acids (NEFA) were decreased, by glucose treatment. During the first 2 h after feed distribution, plasma insulin increased, and plasma glucose and NEFA decreased, in both control and treated cows. Afterward, plasma glucose increased in treated cows but further decreased in control cows. The difference reached 8 mg/100 ml without any change in plasma insulin. During the meal, concentrations of growth hormones in plasma were inhibited to a similar extent in both groups. In response to intravenous glucose or insulin challenges, changes in plasma glucose, NEFA, and insulin stimulated by glucose were also very similar in both groups. In conclusion, duodenal infusion of glucose increased basal plasma concentrations of glucose and insulin, increased postprandial plasma glucose, and decreased NEFA without inducing insulin resistance. Glucose treatment did not change milk yield but decreased milk fat yield, mainly through a decrease in the yield of C18 fatty acids that were derived from circulating fatty acids. In the absence of insulin resistance, the decrease in the yield of C18 fatty acids might be attributed to an inhibition of adipose lipolysis or an increase in adipose lipogenesis. PMID- 9406079 TI - Effect of anionic salts in prepartum diets based on alfalfa. AB - This study compared prepartum diets based on grass, alfalfa, or alfalfa and anionic salts to investigate their effect on Ca metabolism, acid-base status, endocrine response, disease incidence, and lactational performance of periparturient dairy cows. Forty-five nonlactating Holstein cows in their last 3 wk of gestation were fed a control diet based on grass hay with a dietary cation anion difference [expressed as milli-equivalents of ((Na + K) - (Cl + S))/100 g of dietary dry matter] of +30 or diets based on alfalfa with a dietary cation anion difference of either +35 or -7. Cows fed the diet with the dietary cation anion difference of -7 had the lowest urine pH prepartum and had the highest concentrations of ionized Ca in blood and total Ca in serum at parturition. Increases in 1,25-(OH)2 vitamin D per unit decrease in total Ca in serum were greatest for cows fed the diet with a dietary cation-anion difference of -7. Also, cows fed this same diet consumed the most dry matter postpartum. Incidences of health disorders were 13% (10 of 75), 12% (9 of 75), and 5% (4 of 75) for cows fed the diets with dietary cation-anion differences of +30, +35, and -7, respectively. Results indicate that alfalfa, when supplemented with anionic salts, is a viable forage for prepartum dairy cows. PMID- 9406080 TI - Timothy grass or alfalfa silage for cows in midlactation: effect of supplementary barley. AB - Twenty-four multiparous Holstein cows averaging 104 d of lactation were used in a trial with a split-plot design to evaluate the nutritive value of two silages, timothy grass or alfalfa, both treated with formic acid and stored in plastic bag silos. Silages were offered for ad libitum intake either alone or with 17 or 34% (dry matter basis) dry-rolled barley. Both silages contained similar amounts of acid detergent fiber (ADF) (27.5 and 26.7% for timothy grass and alfalfa, respectively). After 110 d of storage, alfalfa silage contained higher amounts of organic acids and NH3 N but had lower soluble N. Total dry matter intake (DMI) and silage DMI were similar between cows fed both silages. Increased barley proportion decreased silage DMI (19.2 to 14.2 kg/d). Apparent total tract digestibility of dry matter was unaffected by treatment. The digestibility of neutral detergent fiber and ADF was higher for the timothy grass silage than for alfalfa silage and was unaffected by the barley percentage added to either silage. Milk yield was lower (23.9 to 22.6 kg/d) for cows fed the highest proportion of barley. The 4% fat-corrected milk yield was unaffected by treatment. Percentages of fat, protein, and total solids in milk were higher for cows fed diets with the higher barley content. Milk fat and protein yields were similar among treatments. Urea in blood was lower for cows fed timothy grass silage than for cows fed alfalfa silage (4.68 vs. 6.23 mg/100 ml). These results suggest that timothy grass silage and alfalfa silage, when stored at a similar ADF content, have comparable nutritive value for midlactation cows. PMID- 9406081 TI - Comparison of barley, hull-less barley, and corn in the concentrate of dairy cows. AB - Twelve multiparous and 12 primiparous lactating Holstein cows were used to compare the effects of hull-less barley with barley and corn on dry matter intake (DMI), digestibility, and milk production. Three concentrates were formulated using steam-rolled grains: barley, hull-less barley, or corn. During three 21-d periods, cows received a total mixed diet consisting of 60% concentrate, 30% barley silage, and 10% cubed alfalfa hay [dry matter (DM) basis]. Milk production and DMI were higher for cows fed the corn diet than for cows fed the barley or hull-less barley diets; no interaction with parity was detected. The DMI of cows fed the hull-less barley and barley diets were similar. Despite the higher estimated energy density of the hull-less barley diet, milk production was similar for cows fed the hull-less barley and barley diets because of the lower digestibility of the hull-less barley. Results of an in situ study showed that, for steam-rolled grains, DM and starch from hull-less barley were less degradable than were DM and starch from barley, although the opposite result was observed for ground grains. For steam-rolled hull-less barley, low ruminal degradabilities of DM and starch were apparently not compensated by high intestinal digestibility because total tract digestibility and milk production were lower than expected. Although the net energy for lactation value of hull-less barley is higher than that for barley, milk production by cows might be limited unless hull-less barley is adequately processed to ensure high ruminal and total tract digestibilities. PMID- 9406082 TI - Wet tomato pomace ensiled with corn plants for dairy cows. AB - The fermentation characteristics and nutritive value of a mixture of whole corn plants and wet tomato pomace were studied. Laboratory silos were filled with mixtures of corn plant dry matter (DM) and tomato pomace DM in the following ratios: 100:0, 94:6, and 88:12. The initial pH of the mixtures decreased linearly as the amount of tomato pomace increased, but no treatment effects were found after 3 d of ensiling. At d 56, concentrations of lactic acid and nonprotein N (as a percentage of total N) decreased linearly as tomato pomace increased. Whole corn plants were chopped from a single field, mixed with 0 or 12% tomato pomace (percentage of DM), and ensiled in two concrete stave silos. The addition of tomato pomace increased concentrations of crude protein (7.6 to 9.8%), lignin (2.4 to 6.1%), and fatty acids (2.5 to 3.8%). Both silages were mixed into an isonitrogenous total mixed ration that contained 40% concentrate. The total mixed rations were fed to lactating dairy cows for 60 d. Milk production (35.5 kg/d), milk composition, DMI (22.7 kg/d), nutrient digestibility, and N balance were not affected by the addition of tomato pomace. Wet tomato pomace can be blended with corn plants before ensiling without greatly altering fermentation. The nutritional value of the mixture was equivalent to that of untreated corn silage. PMID- 9406083 TI - Substitution of neutral detergent fiber from forage with neutral detergent fiber from by-products in the diets of lactating cows. AB - Four lactating dairy cows that were ruminally and duodenally cannulated were used in an experiment with a 4 x 4 Latin square design to determine the effects of the substitution of neutral detergent fiber (NDF) from forage with NDF from wheat middlings, corn gluten feed, or a blend of distillers dried grains and hominy. Dietary crude protein and NDF averaged 18 and 31%, respectively, for the diet with 71.2% of the NDF from forage (control diet) and for diets with 55% of the NDF from forage (by-product diets). The substitution of NDF from these by products for forage NDF did not affect dry matter intake (20.1 kg/d) or digestibility of organic matter. Total tract digestibility of acid detergent fiber was lower for cows fed the diet containing a blend of distillers dried grains and hominy than for cows fed the diet containing corn gluten feed. Microbial crude protein synthesis, milk production (23.9 kg/d), and milk fat percentage were similar for all cows, regardless of diet. Cows fed the diets containing wheat middlings or a blend of distillers dried grains and hominy had reduced ruminal pH compared with that of cows fed the diet containing corn gluten feed or the control diet. Diets containing 55% of total NDF from forage with 31% of total NDF from corn gluten feed, wheat middlings, or a blend of distillers dried grains and hominy can supply sufficient effective fiber to maintain normal ruminal function. PMID- 9406084 TI - Influence of fat source and sorghum grain treatment on performance and digestibilities of high yielding dairy cows. AB - Forty-eight lactating Holstein cows averaging 81 d in milk were allotted to eight blocks based on milk yield during the 14-d pretreatment period and randomly assigned to six treatment groups in a 2 x 3 factorial arrangement of treatments for 64 d. Factors were type of sorghum grain processing [dry-rolled vs. steam flaked; fed at 34% of dry matter (DM) in a total mixed ration (TMR) based on alfalfa] and type of supplemental fat (2.5% of DM as cottonseed oil, tallow, or prilled fatty acids). Compared with dry-rolled sorghum, steam-flaked sorghum did not affect milk yield, fat percentage, or fat yield but did increase milk protein percentage, body weight gains, and estimated net energy for lactation (22%). Fat source did not affect lactational response, but, compared with tallow, prilled fatty acids tended to decrease DM intake. Steam-flaked sorghum, compared with dry rolled sorghum, increased digestibilities of DM, organic matter, crude protein, and starch, regardless of fat source. The TMR containing prilled fat had lower digestibilities of DM and organic matter than did TMR containing cottonseed oil or tallow; and TMR containing prilled fat had lower digestibilities of crude protein and total fatty acids than did TMR containing tallow. This study showed that steam-flaking of sorghum grain increased milk protein content, body weight gains, and estimated net energy for lactation, regardless of dietary fat source. PMID- 9406085 TI - Dietary soybeans extruded at different temperatures: milk composition and in situ fatty acid reactions. AB - Twenty-four multiparous Holstein cows, 4 of which were ruminally fistulated, were assigned to one of four diets containing full-fat soybeans, either raw or extruded at 120, 130, or 140 degrees C. Our hypothesis was that the extrusion of full-fat soybeans, as well as the extrusion temperature, would affect the bypass of fatty acids in the rumen and, thus, would modify the fatty acid profile of milk fat. Total mixed diets containing 23.7% soybeans (percentage of DM) were fed for 8 wk. Milk yield was lower, and the proportion of milk CP was higher, for cows fed raw soybeans than for cows fed extruded soybeans. Compared with raw soybeans, extruded soybeans increased the concentration of delta-11-trans-C18:1 from 2.72 to 11.41% in milk fat but had no effect on yield or percentage of milk fat. Polyunsaturated fatty acids of raw soybeans disappeared more rapidly than did those of extruded soybeans from bags incubated in the rumen of fistulated cows. However, more delta-11-trans-C18:1 and C18:0 appeared in bags containing extruded soybeans than in bags containing raw soybeans. Extrusion of full-fat soybeans influenced the metabolism of fatty acids in the rumen and the fatty acid profile of milk fat, but the temperature of extrusion had only minor effects on these parameters. PMID- 9406086 TI - Comparison of in situ and in vitro techniques for measuring ruminal degradation of animal by-product proteins. AB - Ruminally undegraded protein (RUP) values of blood meal (n = 2), hydrolyzed feather meal (n = 2), fish meal (n = 2), meat and bone meal, and soybean meal were estimated using an in situ method, an inhibitor in vitro method, and an inhibitor in vitro technique applying Michaelis-Menten saturation kinetics. Degradation rates for in situ and inhibitor in vitro methods were calculated by regression of the natural log of the proportion of crude protein (CP) remaining undegraded versus time. Nonlinear regression analysis of the integrated Michaelis Menten equation was used to determine maximum velocity, the Michaelis constant, and degradation rate (the ratio of maximum velocity to the Michaelis constant). A ruminal passage rate of 0.06/h was assumed in the calculation of RUP. The in situ and inhibitor in vitro techniques yielded similar estimates of ruminal degradation. Mean RUP estimated for soybean meal, blood meal, hydrolyzed feather meal, fish meal, and meat and bone meal were, respectively, 28.6, 86.0, 77.4, 52.9, and 52.6% of CP by the in situ method and 26.4, 86.1, 76.0, 59.6, and 49.5% of CP by the inhibitor in vitro technique. The Michaelis-Menten inhibitor in vitro technique yielded more rapid CP degradation rates and decreased estimates of RUP. The inhibitor in vitro method required less time and labor than did the other two techniques to estimate the RUP values of animal by-product proteins. Results from in vitro incubations with pepsin.HCl suggested that low postruminal digestibility of hydrolyzed feather meal may impair its value as a source of RUP. PMID- 9406087 TI - Effect of ruminal degradability of crude protein and nonstructural carbohydrates on the efficiency of bacterial crude protein synthesis and amino acid flow to the abomasum of dairy cows. AB - Four lactating Israeli Holstein cows that were ruminally and abomasally cannulated were used in an experiment with a 4 x 4 Latin square experimental design to study the effects of different amounts of ruminally degradable crude protein (CP) and ruminally degradable nonstructural carbohydrates on ruminal fermentation and efficiency of bacterial CP synthesis. Four diets were formulated to contain the following percentages (percentage of respective fraction) of ruminally degradable protein (RDP) and ruminally degradable nonstructural carbohydrates, respectively: 1) 73.8 and 85.3%, 2) 72.4 and 75.3%, 3) 67.7 and 86.0%, and 4) 66.3 and 76.0%. The 2 x 2 factorial effects of high and low concentrations of RDP or nonstructural carbohydrates were examined. Intakes of DM and organic matter (OM) were similar among treatments, and apparent and true ruminal digestibilities of OM were also similar. Apparent digestibility of CP in the total tract was higher for diets containing high concentrations of ruminally degradable nonstructural carbohydrates. Efficiency of microbial CP synthesis was higher for diets supplemented with low concentrations of RDP and averaged 196 g of microbial CP/kg of OM truly digested in the rumen. Total and bacterial CP flows were higher for diets containing low concentrations of RDP. Therefore, greater amounts of amino acids (AA) of bacterial origin reached the abomasum. The abomasal flow of AA was higher for diets containing low concentrations of RDP. Most of the profiles for essential AA in the abomasum were influenced and balanced by profiles for bacteria. When diets contained a high concentration of RDP (73% of total dietary CP), the supplementation of a high concentration of ruminally degradable nonstructural carbohydrates had no positive influence on bacterial yield or efficiency of bacterial CP synthesis. Other factors, such as AA and peptides included in the RUP fraction, may be important to maximize the efficiency of bacterial CP synthesis. PMID- 9406088 TI - Yield response of lactating Holstein dairy cows to dietary fish meal and bone meal. AB - Experiments were conducted to examine the effects of the source and amount of dietary protein on yield and composition of milk from Holstein dairy cows. Study 1 used 36 multiparous cows at 125 +/- 59 d in milk in, a replicated 2 x 2 Latin square design. Treatments were diets formulated to contain 16% crude protein (CP) in which 11% was fish meal or meat and bone meal supplied 11% of dietary CP. Intakes of dry matter, CP, and net energy for lactation; yields of milk; and percentage of milk fat were not affected by treatment. Fish meal increased contents of milk total N, casein N, and noncasein N but did not increase contents of NPN; fish meal also tended to increase milk CP yields. Study 2 used 78 cows (31 primiparous) at 31 +/- 2 d in milk in a randomized block design. Two treatment diets were formulated to contain 16 or 18.5% CP, and soybean meal was the sole source of supplemental protein in those diets. The two other treatment diets were, formulated to contain 16% CP; in these diets, fish meal or meat and bone meal partially replaced soybean meal. Treatments did not influence yield or composition of milk from multiparous cows. Compared with a soybean meal diet containing 16% CP, a soybean meal diet containing 18.5% CP or diets containing 16% CP and containing meat and bone meal or fish meal increased the milk, yield of primiparous cows similarly. Fish meal or meat and bone meal increased the efficiency of protein utilization for milk yield. PMID- 9406089 TI - A statistical evaluation of animal and nutritional factors influencing concentrations of milk urea nitrogen. AB - Data from 35 trials with 482 lactating cows fed 106 diets were used to study the effects of animal and dietary factors on the relationship between milk and blood urea N and the value of milk urea N in the assessment of protein status. In two trials, urea N in whole blood and in blood plasma were closely related (r2 = 0.952); the slope was not significantly different from 1.0, and the intercept was not significantly different from 0. Regression of milk urea N on blood urea N with a mixed effects model using all 2231 observations yielded the equation: milk urea N (milligrams of N per deciliter) = 0.620 x blood urea N (milligrams of N per deciliter) + 4.75 (r2 = 0.842); this model accounted for a significant interaction of cow and blood urea N. Single factors that yielded significant regressions on milk urea N with the mixed effects models were dietary crude protein (CP) (percentage of dry matter; r2 = 0.839), dietary CP per megacalorie of net energy for lactation (NEL) (r2 = 0.833), excess N intake (r2 = 0.772), N efficiency (r2 = 0.626), and ruminal NH3 (r2 = 0.574). When all factors were analyzed at once, 12 were significant in a mixed effects model. Blood urea N, body weight, yield of fat-corrected milk, dietary CP content, excess N intake, dry matter intake, and days in milk were positively related to milk urea N, and parity, milk and fat yield, dietary CP per unit of NEL content, and NEL intake were negatively related to milk urea N. In one trial, the mean urea concentration was 35 times greater in urine than in milk; lower proportions of total urea excretion in milk were observed in the a.m. sampling (1.8%) than in the p.m. sampling (3.3%). Measuring urea N in a composite milk sample from the whole day substantially improved reliability of data. The number of cows fed a specific diet that must be sampled to determine mean milk urea N within 95% confidence intervals with half widths of 1.0 and 2.0 mg of N/dl was estimated to be 16 and 4, respectively. PMID- 9406090 TI - Effects of lasalocid in milk replacer of calf starter on health and performance of calves challenged with Eimeria species. AB - Holstein bull calves (n = 48) were purchased from local sale barns at 3 to 7 d of age and were assigned randomly to a 2 x 2 factorial arrangement of lasalocid in milk replacer (0 or 80 mg/kg) and in calf starter (3 or 44 mg/kg of dry matter). On d 10 after arrival, calves were orally dosed with 100,000 Eimeria oocysts. Intakes of calf starter and milk replacer, body weight (BW), BW gain, excretion of fecal oocysts, and fecal scores were determined. Calves fed lasalocid in milk replacer consumed more calf starter, had greater BW gain, shed fewer oocysts in feces, and scoured less frequently and less severely than did calves fed no lasalocid or those fed lasalocid in calf starter alone. The combination of lasalocid in milk replacer and in calf starter did not improve performance above that of calves fed lasalocid in milk replacer alone. Low intake of calf starter prior to weaning may provide an insufficient amount of lasalocid to control effectively the effects of coccidiosis when calves are infected with Eimeria at an early age. Use of coccidiostats in milk replacers may reduce the effects of coccidiosis in young calves that are infected with Eimeria at an early age. PMID- 9406091 TI - A comparison of milk protein sources in diets of calves up to eight weeks of age. AB - Four dietary ratios of dried skim milk and whey protein concentrate as the primary protein sources in milk replacers were evaluated using digestibility, growth performance, health characteristics, and blood measurements of calves that were 12 to 36 h of age to 8 wk of age. Sixteen Holstein bull calves, housed individually within an environmentally controlled facility, were randomly allotted to diets. Protein sources in the milk replacer were expressed as ratios (percentages) of dried skim milk to whey protein concentrate in diet 1, 100:0; diet 2, 67:33; diet 3, 33:67; and diet 4, 0:100. There were no significant differences in the mean number of days that calves scoured. Apparent biological value and N retention differed among diets over all weeks. Apparent digestibilities and biological values (percentages) over all weeks were diet 1, 82.50 and 67.71; diet 2, 82.85 and 70.15; diet 3, 83.82 and 67.46; and diet 4, 84.03 and 72.25, respectively. Diets varying in the ratio of dried skim milk to whey protein concentrates had no effect on health, growth, or apparent digestibilities in Holstein calves up to 8 wk of age. PMID- 9406092 TI - Effects of interactions between type and milk production on survival traits of Canadian Holsteins. AB - Effects of the interaction between type and production on two measures of functional herd life were examined for Canadian Holsteins. Data were records of survival through first lactation for 1,153,706 cows and number of lactations initiated (maximum of five lactations) for 705,930 cows. Survival data were regressed on ETA for type traits of the sire of each cow after the cows were assigned to groups with low, medium, or high production. Survival through first lactation was analyzed with a threshold model. Factors in the model included herd year-season; age at calving; month of calving; interaction of registry status, change in herd size, and season; fat and protein production; and linear regressions of sire ETA for type within each production class. Numbers of lactations were analyzed with a linear model that also included month of last calving. Overall conformation and udder traits had the largest effects on survival through first lactation. Effects on number of lactations for feet and leg traits were about the same as for udder traits. Interactions were significant. Type traits were relatively unimportant for herd life of low producing cows. Few differences were observed in the relationships between herd life and type for medium versus high producing cows, indicating no need to increase the emphasis on type in response to current trends for greater production. PMID- 9406093 TI - Marked effect of beta-lactoglobulin polymorphism on the ratio of casein to total protein in milk. AB - The relationship between genetic variants for milk protein and the composition of milk was analyzed on 4475 repeated milk samples from individual cows; 371 dairy cows of the Swedish Red and White breed and 204 cows of the Swedish Holstein breed were used. The registrations included percentages of casein, protein, fat, and lactose in combination with milk yield and SCC. The genotype of individual cows for alpha(s1)-CN, beta-CN, kappa-CN, and beta-LG was determined by alkaline and acidic PAGE. A mixed animal model was used for the analysis; beta-LG and aggregate casein genotypes were included simultaneously as separate fixed effects in the statistical model. The results suggest a positive additive effect of the beta-LG B allele on casein content and on the ratio of casein to total protein. For the latter trait, the beta-LG genotype accounted for a relatively large part of the phenotypic variance, corresponding to a reduction in residual variance of 11% when included in the model. The corresponding value for casein content was 0.5%. The lack of unfavorable associations between milk protein variants and the traits included in this study makes the beta-LG gene an obvious candidate when the breeding objective is improved conversion of milk protein into cheese. PMID- 9406094 TI - Genetic correlations between longevity and conformation traits in an upgrading dairy cattle population. AB - Genetic correlations between longevity and conformation traits were estimated using data on Dutch Black and White cows born in 1978 (11,558 records), 1982 (39,252 records), and 1989 plus 1990 (58,864 records). Longevity traits considered were number of lactations, herd life, and stayabilities until 36 and 48 mo of age and their functional equivalents (i.e., the longevity traits corrected for production). For the 1989 plus 1990 data file, only stayabilities until 36 and 48 mo of age were considered. Conformation traits were rear legs set, front teat placement, udder depth, suspensory ligament, and subjective scores for udder, feet and legs, and type. Also investigated was a possible nonlinear relationship between conformation and longevity traits. Genetic correlations between conformation and longevity traits differed between years of birth, mainly because farmers practiced large-scale upgrading with Holstein Friesian bulls during the period considered, which caused a change in desired type. Therefore, the predictive value of conformation traits for longevity based on data from an upgrading population, might be limited. Estimates of genetic parameters should be based on the most recent data possible, and these parameters should be reestimated over time. From the 1989 plus 1990 data file, subjective scores for udder and feet and legs had the highest predictive values for functional longevity. Quadratic relationships between conformation and longevity traits did exist, but generally the linear relationships prevailed. PMID- 9406095 TI - Lactation yields and accuracies computed from test day yields and (co)variances by best production. AB - Lactation records are calculated from data on milk, fat, and protein obtained from one or more milkings on several days during the lactation. The test interval method, which estimated missing daily milk yields by simple interpolation, was used for many years for standard monthly data but may not be as useful for the wider variety of test plans now being proposed. More accurate 305-d yields can be computed using best prediction, which has optimum properties if means and (co)variances are known and distribution is multivariate normal. The covariance of test day and 305-d yields is multiplied by the inverse of the test day (co)variance matrix, which is then multiplied by the test day deviation vector. This predicted 305-d deviation plus the mean 305-d yield equals the predicted 305 d yield. Similar algebraic methods are used to compute the correlation of true and estimated 305-d yields, which is needed to calculate lactation weights. Computation times were affordable but not trivial; they ranged from 0.001 to 1 s per lactation. Equations were modified to account for differing accuracies of data for partial days, means for multiple days, and data for unsupervised tests. Complete or incomplete lactations recorded with very different testing plans can be graphed and compared by best prediction. PMID- 9406096 TI - Lactation curves of Valle del Belice dairy ewes for yields of milk, fat, and protein estimated with test day models. AB - Test day models were used to estimate the lactation curves for Valle del Belice ewes and to study the main environmental effects on milk yield and on percentage of fat and protein. Environmental effects were treated as fixed. A random effect was associated with each lactation to evaluate the mean correlation among all test day records of an individual ewe. Lactation curves were constructed by adding solutions for classes of either days in milk nested within parity or days in milk nested within season of lambing to appropriate general means. Parity primarily affected the lactation curve for milk yield, which was lower and flatter for first lambing ewes; effects on fat and protein were smaller. Season of lambing affected all traits. Seasonal productivity had the greatest effect on milk composition, resulting in an imbalance between fat and protein percentages. Flock and feed supplementation affected only the lactation curve for milk yield. The lactation curve of Valle del Belice ewes stood at a relatively high level. However, the presence of notable, perturbative effects (environmental and random variation) on milk yield and composition suggests that management is unable to meet the requirements of ewes consistently. PMID- 9406097 TI - Effects of season, herd size, and geographic region on the composition and quality of milk in the northeast. AB - Our objectives were to describe the milkshed comprising herds in New York, western New Jersey, and central and eastern Pennsylvania in regard to milk yield, composition, and quality and also to estimate the effects of season, herd size, and geographic area on those same variables. Data were collected from July 1993 through June 1994 from 3450 herds. The effect of a somatic cell count (SCC) limit of 500,000/ml on milk yield and the composition of monthly bulk tank milk for all marketed milk was estimated as was the frequency of deliveries of milk that contained SCC that were greater than this limit. All general linear models for mean monthly yield of milk and milk components (fat and protein) and SCC were significant for fixed effects of month and herd size within quartiles for herd size (defined by the number of lactating cows) and significant absorbed effects of herds within quartiles for herd size within subregion. Milk yield, milk components (kilograms), true protein percentage, and SCC were significantly higher in spring than in fall for both data files (complete data file and data file containing only herds with SCC < 500,000/ml). Thirty-five percent of herds with < 27 lactating cows but only 15.3% of herds with > 62 lactating cows had > or = 1 mo with an SCC > 500,000/ml. For herds in the subregions, percentages of shipments with an SCC > or = 500,000/ml ranged from 10.5 to 20.2%. Herds with < 27 lactating cows contributed to the milkshed a disproportionate percentage of SCC (11%) compared with their percentage of contribution of milk (5%). PMID- 9406098 TI - Evaluation of selected antibiotic residue screening tests for milk from individual cows and examination of factors that affect the probability of false positive outcomes. AB - Total composite milk samples from 131 cows in one herd were analyzed. Eight beta lactam residue screening tests were evaluated for performance using milk from individual cows and factors that affect the rate of false-positive outcomes were determined. Cows were not treated with an antibiotic for at least 30 d prior to sampling. Tests evaluated were Delvotest P, Charm Cowside, Charm Farm, Penzyme, Valio T101, LacTek, CITE Probe, and Charm Bacillus stearothermophilus disk assay. Cows averaged 155 d of lactation. Milk production at the time of sampling ranged from 3.6 to 26.3 kg per milking per cow. The somatic cell count of milk averaged 243 x 10(3)/ml and ranged from 8.5 x 10(3)/ml to 3437 x 10(3)/ml. Total viable bacteria counts averaged 197.8 x 10(3)/ml. Total coliform counts ranged from 0 to 205/ml. Selectivity rates (rate of truly negative samples that were found to be negative by the assay) were greater than 90% for all tests except the CITE Probe test. Use of logistic regression showed that an increase in colony-forming units was associated with a decrease in the probability of a false-positive outcome for the CITE Probe test. Milk production, coliform counts, and parity each affected the probabilities of positive outcomes for different tests. Except for one test, selectivity rates of the beta-lactam residue screening tests for milk from individual cows was greater than 0.9. PMID- 9406099 TI - Recovery of Staphylococcus aureus from centrifuged quarter milk samples. AB - The identification of cows that are positive for mastitis caused by Staphylococcus aureus is difficult under field conditions. The frequency of isolation of S. aureus from quarter milk samples was compared with the frequency of recovery of S. aureus from sediment after centrifugation of those same samples. Overall, 776 quarter milk samples from 194 cows were studied. Cultures that were positive for S. aureus were obtained from 82 samples; 153 sediments from quarter milk samples were also positive for S. aureus. The results of this investigation showed that cultures of the sediment of quarter milk samples increased the number of positive outcomes up to 145.5%, depending on the herd. Using a different group of samples, including samples taken 1 to 5 d or 7 to 10 d after calving and samples taken after intramammary therapy, a 94% increase in cultures that were positive for S. aureus after centrifugation was found compared with cultures of the same quarter milk samples that were not centrifuged. Sedimented cultures may be useful in S. aureus control programs that require the segregation, selective treatment, or culling of cows that are positive for S. aureus. PMID- 9406100 TI - Animal by-products contaminated with Salmonella in the diets of lactating dairy cows. AB - As part of a total mixed ration, two rumen-fistulated dairy cows were fed meat and bone meal that had been artificially contaminated with Salmonella spp. Samples from the rumen, feces, and milk were taken 3 d/wk and cultured for salmonella. Rectal temperatures and rumen pH were also measured at the time of sample collection. Over the 2-mo study, salmonella were intermittently recovered from rumen contents, from feces, and from necropsy specimens of rumen contents, cecal contents, and mesenteric lymph nodes. No excretion of salmonella in milk was detected. An elevated rumen pH was associated with increased isolation of salmonella. No clinical illness was observed for either cow. Meat and bone meal that has been contaminated with low concentrations of salmonella is unlikely to result in clinical illness in healthy adult lactating cows. However, dairy producers should continue to be concerned about feed biosecurity and water contamination of animal by-products to prevent and control contamination by salmonella. PMID- 9406101 TI - The effect of artificial insemination once versus twice per day. AB - The objective of this study was to determine the effect of inseminating Jersey cows and heifers once per day or according to the a.m.-p.m. rule. A total of 337 artificial inseminations (AI) were completed by three technicians at the University of Tennessee Dairy Experiment Station at Lewisburg for 6 mo. Cows and heifers were inseminated at estrus using the a.m.-p.m. rule on even days of the month. On odd days of the month, AI were once daily between 0800 and 1200 h. Estrus detection was conducted two to three times daily. Pregnancy was confirmed by rectal palpation 60 to 80 d after AI. Herd DHIA averages were a 12.2-mo calving interval, 76 d to first AI, 83% observed estruses, and a 50% conception rate during the trial. Pregnancy data were analyzed with a model including treatment, AI, lactation number, parity, technician, and group. This study grouped cows and heifers according to when they were in estrus and inseminated (a.m.-a.m., a.m.-p.m., or p.m.-a.m.); means were 43.7, 57.9, and 59.0%, respectively. The a.m.-p.m. AI versus once per day AI yielded a pregnancy rate of 55.6% versus 51.3%. These results show no difference among Jersey cows or heifers that were inseminated artificially once daily in the a.m. However, those cows and heifers inseminated in the a.m. of first estrus detection had a lower pregnancy rate. PMID- 9406103 TI - Body measurements and body weights of special-fed Holstein veal calves. AB - Changes in various body dimensions of special fed veal calves were measured and correlated with body weight (BW) at three specific times during the growth period as contemporaries and over the entire feeding period as noncontemporaries. The calves (n = 826) were weighed and measured for body length, heart girth, wither height, and hip width at 2, 8, and 16 wk after arrival at the veal farms. Each of the four measurements, expressed as ratios to BW, decreased over the feeding period; decline in the ratio of hip width to BW was less than the decreases in the other ratios. Linear models to predict contemporary BW within each age group based on all body measurements were developed; R2 values for models for 2, 8, and 16 wk were 0.72, 0.77, and 0.76, respectively. Within each of the three age classes, a model including linear, quadratic, and cubic terms of heart girth yielded the highest R2 values of any single measurement (0.46, 0.63, and 0.67 for data for 2, 8, and 16 wk, respectively). The addition of heart girth as a second linear measurement to three-term models containing only one other measurement increased the R2 more than did the addition of any other single linear expression, except for the equation based on body length. When all records on all calves were combined and the observations were treated as noncontemporaries, the R2 was 0.97 for a linear model that included all four measurements. However, this R2 was essentially the same as the R2 from a three-term model using only heart girth. The cubic models in descending order of R2 values were heart girth, body length, hip width, and wither height. These results suggest that BW can be predicted accurately in a group of noncontemporary male veal calves ranging from 2 to 16 wk after the start of the feeding period. However, the BW of calves within contemporary groups (2, 8, and 16 wk) cannot be predicted accurately according to R2 values. PMID- 9406102 TI - Sperm numbers inseminated in dairy cattle and nonreturn rates revisited. AB - Three experiments were conducted to test fertility when sperm numbers per insemination ranged from 10 x 10(6) to 40 x 10(6) total sperm. All semen was from Holstein bulls that were on a regular schedule of semen collection. The semen was extended with heated homogenized whole milk, cooled, glycerolated, and frozen according to standard procedures. Semen was distributed to a large group of inseminators to minimize differential field effects on treatment. All experiments were a randomized block design, including a split plot in Experiment 2. In Experiment 1, data for 31,399 first inseminations distributed among treatments of 20 x 10(6), 25 x 10(6), 30 x 10(6), and 40 x 10(6) total sperm resulted in 69.8, 70.0, 70.1, and 70.1% nonreturns at 59 d, respectively. In Experiment 2, data for 18,197 first inseminations divided over treatments of 12 x 10(6), 16 x 10(6), and 20 x 10(6) total sperm resulted in 70.2, 72.4, and 70.8% nonreturns at 59 d, respectively. In Experiment 3, 38,890 first inseminations distributed over treatments of 10 x 10(6), 13 x 10(6), 16 x 10(6), and 20 x 10(6) total sperm resulted in 70.5, 72.2, 73.1, and 71.5% nonreturns at 59 d, respectively. Bull nonreturns ranged from 64 to 76% in the three trials. These results indicate that, under good conditions, total sperm numbers per straw can be reduced to 10 x 10(6) total sperm with a reduction of nonreturn rates at 59 d, for most bulls, of about 1 percentage unit from the maximum when professional inseminators are use. PMID- 9406104 TI - Variations in the management of cancer in the NHS: a legitimate cause for concern? AB - Wide variations still exist in the UK in the management of common cancers in adults. Comprehensive high-quality audit data are needed to relate variations in outcomes to differences in clinical management. The management of cancer in the elderly is often difficult due to co-morbidity. Access to oncological advice should be available to all patients irrespective of age. Development of and adherence to nationally agreed treatment protocols is a key measure in reducing variations in treatment and in outcomes for patients with cancer. PMID- 9406105 TI - Breast cancer screening in younger women: evidence and decision making. AB - Contrasting conclusions on the efficacy of routine breast cancer screening in younger women, under age 50, have been produced by expert and influential groups, particularly in the United States. In an international workshop in 1993, and again at a consensus development conference in 1997, the National Institutes of Health and the National Cancer Institute concluded that evidence for efficacy was uncertain, and routine screening could not be recommended. The 1997 conference concluded that the individual decision had to be made by each woman and her health care provider. In contrast, the American Cancer Society has advocated routine screening, despite accepting that the randomized trial evidence does not clearly support it. The decision of the 1997 NCI consensus conference has been rejected by the director of the NCI, and a similar controversy occurred in 1993. On two occasions, US Senate subcommittees have affirmed support for screening and criticized the conclusions of expert groups. In this paper, the arguments raised in these discussions, and the differing ways in which scientific evidence has been assessed, are discussed. PMID- 9406106 TI - Interpreting health outcomes. AB - Interest in outcomes is universal. To patients, good outcomes represent their highest hopes for therapy; to health care professionals, good outcomes are the desired end-point of a complex web of care. More recently, politicians and health care managers too have shifted their emphasis away from health service activity and towards what is termed 'health gain'. The rise of the outcomes movement appears irresistible. However, the difficulties in interpreting outcomes data will not go away. Outcomes measured using routine data are subject to numerous biases and many practical difficulties. Despite recent statistical, methodological and technological advances, comparisons of outcomes at best provide us with weak evidence of either the effectiveness or the quality of health care. And sometimes they may frankly mislead. The apparent intuitiveness of outcomes monitoring has broad public appeal. But enthusiasm for outcomes needs to be tempered with a clear understanding of their limitations. PMID- 9406107 TI - Certainty, probability and abduction: why we should look to C.S. Peirce rather than Godel for a theory of clinical reasoning. AB - This paper argues that Godel's proof does not provide the appropriate conceptual basis on which to counter the claims of evidence-based medicine. The nature of, and differences between, deductive, inductive and abductive inference are briefly surveyed. The work of the American logician C.S. Peirce is introduced as a possible framework for a theory of clinical reasoning which can ground the claims of both evidence-based medicine and its critics. PMID- 9406108 TI - Meta-analyses using individual patient data. AB - Systematic reviews of randomized controlled trials often provide the most reliable information on which to base treatment policy. However, to be reliable, such reviews need to contain a high proportion of all the relevant randomized evidence. This relates both to the need to find all trials and the need to analyse data on all participants. One way to achieve this is through a collaboration in which those responsible for the trials supply data on each randomized patient for an individual patient data meta-analysis. However, such projects require more time and resources than more conventional reviews and are still rare. This paper illustrates how they can help to achieve the aim of using complete data in a systematic review. PMID- 9406109 TI - A systems analysis approach to medical error. AB - Evidence from various sources indicates that a substantial number of hospitalized patients suffer treatment-caused injuries. Most of these injuries result from errors. Yet physicians and other health professionals have been reluctant to admit and address the problem of errors, both because of feelings of guilt and from the desire to avoid punishment or disapproval by colleagues. Research in cognitive psychology and human factors has shown that most errors result from defects in the systems in which we work. These are failures in the design of processes, tasks, training, and conditions of work that make errors more likely. Meaningful reduction of errors requires correction of these systems failures. Barriers to reduction of errors include the complexity of health care systems, difficulties in information access, tolerance of stylistic practices, and fear of punishment that inhibits reporting. Most institutions also lack effective methods for detecting and quantifying errors. Significant improvements in error reduction will require major commitments by organizational leadership and the recognition that errors are evidence of deficiencies in systems, not deficiencies in people. PMID- 9406110 TI - Advancing the standards of clinical research: the urgent need for new methods and better data. AB - Some of the weaknesses of the randomized controlled trial (RCT) are pointed out, among them unavoidable biases, the possibility of coming to erroneous conclusions by the 'luck of the draw' and the fact that most such experiments lack a doctor/patient relationship and are of little assistance in clinical decision making. An exaggerated propensity on the part of the medical profession to regard the RCT as the only 'scientific' method for research into therapeutics is noted. The potential advantages of post-marketing surveillance of new interventions in a physician-centred manner are outlined. Such a data base could ultimately be expanded into a data base that records all activity in a given practice. The PACE strategy, employed by haematologists in the north of England, and the sentinel practice primary care research networks used by family doctors contain elements of the system proposed here. However, each possesses disadvantages that limit its applicability to special cases. Random choice of practices for any particular observational study of a new intervention could lend it some of the features of an experiment. PMID- 9406112 TI - The study of forests and ecosystems: how to grow better trees: a commentary on performance evaluation and improvement strategies in health care. PMID- 9406111 TI - Medical fallibility and the autopsy in the USA. AB - The theory of 'necessary fallibility', originated by Gorovitz & MacIntyre (1976, In Science, Ethics and Medicine, Hastings Center, NY), explains a major unfamiliar reason for unavoidable errors in medicine. A brief historical review is presented of autopsy studies assessing the accuracy of clinical diagnoses, and of changing perceptions of the public and physicians leading to the current misconception in the USA that all errors are avoidable. It is suggested that a prospective autopsy study is needed to test the theory of 'necessary fallibility'. Validation of this theory would benefit an understanding of medical fallibility by the public and physicians, and challenge current practices in the management of malpractice and patient injury. PMID- 9406113 TI - Evidence-based everything. AB - Academic medicine often seems to be swayed more by fashion than science. Establishment team consensus is only needed where there is ambiguity. Evidence based medicine is a new term for informed decision making and facilitated learning is purported to do away with authoritarian indoctrination. Problems that arise from the emphasis on team decisions, evidence based medicine and facilitated learning are discussed. PMID- 9406114 TI - Nonadrenergic-noncholinergic relaxations of isolated circular muscle from South American opossum esophagogastric junction: is nitric oxide the inhibitory mediator? AB - Nonadrenergic-noncholinergic (NANC) inhibitory nerves are responsible for most of the nerve induced relaxations of gastrointestinal muscle. It has recently been proposed that NANC nerves may release nitric oxide (NO) or a related compound derived from L-arginine. We have recently shown that the South American (SA) opossum is another suitable model to elucidate the mechanism involved in these NANC relaxations. In the present study the effect of NO synthase inhibitors as well as NO inactivators on the NANC-nerve induced relaxations of the circular muscle of the esophagogastric junction (EGJ) of the SA opossum was investigated. It was observed that the NO synthase inhibitors, L-NOARG and L-NAME, caused a concentration-dependent reduction of NANC-nerve induced relaxations which was reversed by L- but not D-arginine. The NO-donors sodium nitroprusside and hydroxilamine as well as NO caused concentration-dependent relaxations of the EGJ circular muscle. In the myenteric plexus of this region, NADPH-diaphorase positive neurons and nerve fibers were observed while in the circular muscle layer only numerous positive fibers were found. The NO inactivators, hydroquinone, pyrogallol and carboxy-PTIO, reduced NO-induced relaxations but failed to affect NANC nerve- and sodium nitroprusside-induced relaxations. Taken together, these findings indicate that NANC nerve induced relaxation of the SA opossum EGJ circular muscle is dependent on neural NO synthase activity and suggest that the neurotransmitter being released is a superoxide resistant molecule, which is unlikely to be the NO radical, or that the activity of NO synthase is required for the release of the actual neurotransmitter rather than for synthesizing the neuromediator. PMID- 9406115 TI - The role of free radicals in the release of noradrenaline from myenteric nerve terminals of guinea-pig ileum. AB - In an attempt to understand the role of free radicals in the regulation of sympathetic neurotransmission, the in vitro secretion of noradrenaline (NA) from synaptosomal preparations of guinea-pig ileum was investigated. Release of endogenous NA was quantified by an electrochemical detection (HPLC-ECD). In the presence of superoxide dismutase (SOD) and catalase at concentrations sufficient to scavenge the free radicals, secretion of NA was attenuated in samples with stimulation of 4-aminopyrine (4-AP) or not (spontaneous release). However, inducing superoxide radicals via the reaction of hypoxanthine with xanthine oxidase failed to modify the secretion of NA, both the 4-AP-stimulated release and the spontaneous secretion. Then, free radicals were induced in synaptosomes using hypoxia-normoxia exposure. Secretion of NA was markedly increased in samples receiving this treatment in a calcium-dependent way because it was attenuated by the removal of calcium chloride from bathing medium. An increase of SOD activity, both Mn-SOD and Cu, Zn-SOD, was also obtained by this exposure. Changes of SOD activities in response to free radicals produced by hypoxia normoxia exposure in ileal synaptosomes can thus be considered. In conclusion, these results suggest that free radicals are formed to involve in the regulation of sympathetic neurotransmission via an increase of calcium influx to enhance the NA release in guinea-pig ileum. PMID- 9406116 TI - Prostaglandin E2 activation of VIP secretomotor neurons in the guinea pig ileum. AB - The effect of prostaglandin E2 (PGE2) on the activity-related expression of the proto-oncogene c-fos in specific populations of enteric neurons was investigated. Segments of guinea-pig ileum were incubated in vitro in the presence or absence of PGE2, and whole mounts of the myenteric and submucosal plexus were prepared for immunocytochemical localization of Fos, VIP and NPY. Control tissues exhibited a low number of Fos-immunoreactive (Fos-IR) neurons (7 +/- 2% of total). Incubation of the tissues with 10-1000 nM PGE2 for 30 min caused a concentration-dependent increase in Fos-IR submucosal neurons (maximum at 100 nM; 39 +/- 6%), which was not inhibited by TTX. PGE2 did not evoke an increase in Fos IR myenteric neurons. In double labeling experiments, Fos colocalized exclusively with VIP in the submucosal plexus, and not with NPY. Exposure of stripped segments of guinea pig ileum in Ussing chambers to 100 nM PGE2 evoked an increase in short circuit current (20 +/- 7 microA/cm2), of which the initial rapid phase could be abolished by TTX, and not by atropine and hexamethonium. It is concluded that PGE2 can activate VIP non-cholinergic secretomotor neurons. PMID- 9406117 TI - Vagal control of left ventricular contractility is selectively mediated by a cranioventricular intracardiac ganglion in the cat. AB - Activation of the vagus nerve leads to decreases in sinoatrial (SA) rate, atrioventricular (AV) conduction, and myocardial contractility. Previous data are consistent with the hypothesis that vagal control of cardiac rate and AV conduction are mediated by two anatomically separated and physiologically independent parasympathetic intracardiac ganglia located in fat pads on the surface of the right and left atria, respectively. These data suggested that vagal control of ventricular contractility might be mediated through another intracardiac ganglion. We examined the ventricles of cat hearts histologically for the presence of ganglia. Multiple small basophilic ganglia composed of a few neurons, and an occasional larger ganglion were found embedded in the epicardial fat surrounding the cranial margin of the anterior surface of the left ventricle, near the juncture with the right ventricle, which we refer to as the CV ganglion. In anesthetized cats, right cervical vagal stimulation decreased SA rate by 44 +/ 5%, decreased the rate of AV conduction by 68 +/- 14%, and reduced ventricular contractility by 19.5 +/- 5.7%. Vagally induced negative inotropism was almost completely prevented by microinjection of a ganglionic blocking drug into the CV ganglion. However, these injections into the CV ganglion did not significantly effect vagally induced decreases in either SA rate or AV conduction. We conclude: (1) that ganglia are found in a fat pad on the surface of the left ventricle of the cat heart and (2) that the CV ganglion selectively mediates the negative inotropic effect of vagal stimulation on the left ventricle. Greater understanding of the physiological functions of intracardiac neuronal circuits may help in developing new strategies to treat disorders of cardiac contractility such as congestive heart failure. PMID- 9406118 TI - Effects of aldehyde/aldose reductase inhibition on neuronal metabolism of norepinephrine. AB - After norepinephrine (NE) is deaminated by monoamine oxidase (MAO), the aldehyde formed is either metabolized to 3,4-dihydroxy-mandelic acid (DHMA) by aldehyde dehydrogenase or is converted to 3,4-dihydroxyphenylglycol (DHPG) by aldehyde or aldose reductase. The present study examined the effects of inhibition of aldehyde and aldose reductase on production of DHPG and DHMA in rats. Mean (+/- S.E.) baseline plasma concentrations of DHPG (4.73 +/- 0.21 pmol/ml) were 60-fold higher than those of DHMA (0.08 +/- 0.01 pmol/ml). Inhibition of aldose and aldehyde reductase reduced plasma DHPG concentrations to 1.88 +/- 0.14 pmol/ml and increased plasma DHMA to 4.43 +/- 0.29 pmol/ml; additional inhibition of MAO reduced plasma DHPG to 0.16 +/- 0.06 pmol/ml and DHMA to 0.19 +/- 0.02 pmol/ml. Inhibition of aldehyde and aldose reductase also increased brain tissue levels of DHMA from 8 +/- 2 to 384 +/- 47 pmol/g and decreased levels of DHPG from 70 +/- 9 to 44 +/- 5 pmol/g. The results show that DHMA is normally a minor metabolite of NE, but becomes a major metabolite after aldehyde/aldose reductase inhibition. PMID- 9406119 TI - Cerebral blood flow in rabbits during the nasopharyngeal reflex elicited by inhalation of noxious vapor. AB - We used chronically implanted Doppler ultrasonic flow probes to measure internal carotid and vertebral blood flow during the nasopharyngeal reflex elicited by inhalation of formaldehyde vapor in conscious rabbits. Internal carotid flow gradually increased to 157 +/- 5% of baseline and vertebral artery increased to 123 +/- 9% of baseline, with maximum values reached approximately 20-40 s after administration of vapor, at a time when arterial pO2 had decreased from 80 +/- 3 to 53 +/- 4 mmHg. Increases in flow were associated with increases in vascular conductance. The delayed increases in cerebral blood flow contrasted with rapid decreases in ear and distal aortic flows, measured at the same time. Our results indicate that forebrain vascular conductance increases in response to inhalation of noxious vapor, possibly reflecting cerebrovascular events associated with hypoxemia. PMID- 9406120 TI - Response to vestibular stimulation of sympathetic outflow to muscle in humans. AB - The objective of the present study was to determine the effect of vestibular stimulation on the sympathetic outflow to muscle in humans. Fourteen healthy volunteers were studied while in the supine position with electrocardiography, blood pressure monitoring and electro-oculography. The muscle sympathetic nerve activity (MSNA) was recorded directly from the bilateral tibial nerves by using microneurographic double recording technique. Caloric vestibular stimulation was loaded by alternate irrigation with 50 ml of cold (10 degrees C) water and 50 ml of hot (44 degrees C) water into the left and right external meatus. After cold water irrigation, two MSNA response peaks were elicited, respectively, before and after the maximum slow phase velocity (SPV) of nystagmus. The first peak of the MSNA enhancement was caused by non-specific factors because its time course coincided with that in cold pressor test with immersion of the subject's hand in ice/water (4 degrees C). Transient suppression of MSNA after cold water irrigation in the period of maximum SPV of nystagmus was observed by cross correlogram analysis between the SPV of the nystagmus and MSNA. After hot water irrigation, only one MSNA response peak was elicited after the period of strong nystagmus. The second peak of MSNA enhancement evoked by cold irrigation (379.4 +/- 221.8%, with the control value set as 100%, mean +/- SE) was significantly higher than that evoked by hot irrigation (243.0 +/- 14.5%). The degree of MSNA enhancement by either cold (the second peak) or hot stimulation was proportional to the maximum SPV of the nystagmus. There was no significant difference between the MSNA responses ipsilateral to and contralateral to the irrigated side. In conclusion, the caloric vestibular stimulation can influence the bilateral sympathetic outflow to muscle in humans. The degree of MSNA enhancement is proportional to the magnitude of vestibular excitement indicated by maximum slow phase velocity of the nystagmus. PMID- 9406121 TI - The nature of detrusor bladder neck dyssynergia in non-neurogenic bladder dysfunction. AB - There have been two major opinions on the pathology or nature of the bladder neck contracture. One is an organic fibrosis, and the other is an accentuated sympathetic nervous function, or detrusor bladder neck dyssynergia. The existence of active detrusor bladder neck dyssynergia in neurogenic bladder was reported in a urodynamical manner using microtip transducer catheters. However, it has not been confirmed whether or not detrusor bladder neck dyssynergia is responsible for bladder neck contracture in patient without neurogenic bladder. The present study was designed to determine by means of video urodynamic study whether or not bladder neck contracture would be of the same nature as detrusor bladder neck dyssynergia in non-neurogenic bladder subjects. The study included 32 male subjects of 16-84 years old (average 52.3): 17 bladder neck contracture subjects including 7 subjects associated with minimum complications (4 with trapped benign prostatic hyperplasia and 3 with incomplete neurological lesion) and 15 non bladder neck contracture subjects (10 healthy volunteers, 2 chronic prostatitis, 3 prostatodynia). A 5-microtip transducer catheter was used to measure the pressure in the bladder and at the bladder neck, the external urethral sphincter and the bulbous urethra during voiding. Proper localization of the transducers was done with an image intensifier. Bladder outlet obstruction localized at the bladder neck (diameters smaller than 0.75 cm) on voiding cystourethrogram was defined as bladder neck contracture. Detrusor bladder neck dyssynergia was defined where pressures were higher at the level of bladder neck than in the bladder during detrusor contraction. An alpha-blocker, terazosin hydrochloride (0.5 mg, b.i.d., two weeks), was orally administered to subjects judged to have detrusor bladder neck dyssynergia by the above methods for the purpose of confirming whether detrusor bladder neck dyssynergia was really due to accentuated sympathetic nervous function. Detrusor bladder neck dyssynergia was found in seven cases with bladder neck contracture: 6 cases with bladder neck contracture with minimum complications and only 1 case with bladder neck contracture without complications (p < 0.01). Detrusor bladder neck dyssynergia was found at the beginning and ending of micturition, but not at maximum flow. In six cases with detrusor bladder neck dyssynergia, the condition disappeared after terazosin. In conclusion, detrusor bladder neck dyssynergia was not thought to be a major factor of voiding dysfunction in bladder neck contracture in non neurogenic bladder. In the presence of sympathetic hyperactivity or in cases with increased number of alphareceptors, detrusor bladder neck dyssynergia occurs, being predominantly noted in trapped benign prostatic hyperplasia and neurological disorder patients. PMID- 9406122 TI - Vagal efferent fibre responses to gastric and oesophageal mechanical and chemical stimuli in the ferret. AB - Gastric and oesophageal afferent inputs to vagal efferent fibres were investigated in Urethane anaesthetized ferrets. Mechanical, chemical, and pharmacological stimuli were tested and efferent activity recorded from single cervical vagal fibres. Fibres showed either no basal discharge or low frequency, irregular patterns of resting discharge; only those which showed > 50% excitation or inhibition of basal activity with both gastric distension and oesophageal balloon distension were studied further. These responses were rapid and maintained only for the duration of the stimuli. 18/32 efferent fibres tested also showed changes in discharge in response to acid infused slowly into the distal oesophagus. These responses were larger after repeated acid infusions. Subsequent intra-oesophageal capsaicin elicited a similar response in 7/8 fibres. These responses were reproducible with repeated capsaicin infusions in 2/4 fibres and desensitized in 2/4 fibres. 2 capsaicin-responsive fibres were unresponsive to oesophageal acidification. 4/12 fibres tested responded to close intraarterial injections of capsaicin and 9/12 to close intraarterial bradykinin. These responses were brief and of short latency. Vagal efferent responses to mechanical and chemical stimuli above were unchanged after the NK-1 receptor antagonist CP96,345 (4 mg/kg i.v.). Subsequently, bilateral vagotomy caudal to the recording site abolished the basal activity in 4/7 fibres. In the 3 fibres where spontaneous activity remained, none of these responded to oesophageal distension or intra-oesophageal acid (2/2 fibres tested) after vagotomy, whereas 2/2 fibres tested still responded to gastric distension. The response of 1 fibre to intraarterial bradykinin and capsaicin was unchanged by vagotomy. We conclude that vagal efferent neurones respond to gastro-oesophageal mechanical inputs and also receive convergent input from oesophageal acid-sensitive and gastrointestinal bradykinin- and capsaicin-sensitive afferents. These afferent inputs are not mediated via NK-1 receptors. There also exists a nonvagal afferent input onto vagal efferent neurones which is probably spinal and likewise non NK-1 receptor mediated. PMID- 9406123 TI - Vagal and sympathetic influences on the ferret lower oesophageal sphincter. AB - This study has investigated the relative involvement of cholinergic, adrenergic, nitric oxide and tachykininergic transmission in extrinsic neural influences on the lower oesophageal sphincter (LOS) in urethane anaesthetized ferrets. A micromanometric assembly (OD 1.75 mm) incorporating a sleeve sensor was used for high-fidelity oesophageal, LOS and gastric pressure measurement at low perfusion rates (< 0.1 ml/min). The LOS response to vagal and splanchnic nerve stimulation (0.5 ms pulse width, 10 s duration) was frequency- and voltage-dependent. LOS responses to stimulation at 20 V, 10 Hz were investigated in separate groups of animals with either L-NAME (100 mg/kg), hexamethonium (15 mg/kg), guanethidine (5 mg/kg), CP96,345 (NK-1 antagonist, 4 mg/kg), atropine (0.4 mg/kg) or propranolol (1 mg/kg). Propranolol treatment was followed by yohimbine (1 mg/kg) and prazosin (0.25 mg/kg). Vagal stimulation caused an immediate decrease in LOS pressure, followed by increase on cessation of stimulation, followed by a prolonged decrease (77 +/- 2%) for up to 5 min. L-NAME did not affect inhibition, but increased excitation 4-fold (p < 0.001). Guanethidine and CP96,345 had no major effect. Hexamethonium decreased the inhibitory (p < 0.05) and excitatory (p < 0.01) responses. Atropine reduced the excitatory response (p < 0.05). Some inhibition still remained if all treatments were combined. Splanchnic stimulation reduced LOS pressure by 70 +/- 6% for 101 +/- 17 s. L-NAME, guanethidine, hexamethonium and CP96,345 all independently significantly reduced inhibition. The combination of guanethidine and CP96,345 usually abolished splanchnic-induced inhibition. Atropine was without effect. Propranolol (1 mg/kg) changed the splanchnic-induced response from mainly inhibition to excitation (100 +/- 44% increase). LOS responses to noradrenaline (1-10 micrograms close IA) showed similar features to responses to splanchnic stimulation. We conclude that vagal stimulation evokes LOS relaxation via activation of established cholinergic and NANC mechanisms and other, unidentified mechanisms. Splanchnic stimulation activates adrenergic neurones probably via nicotinic and non-nicotinic ganglionic mechanisms, which in turn elicit beta adrenergic inhibitory effects on the LOS. Splanchnic stimulation also antidromically activates spinal afferent fibres. These may release substance P from peripheral myenteric plexus and prevertebral ganglionic endings causing activation of myenteric NANC inhibitory neurones and sympathetic neurones, respectively. PMID- 9406124 TI - Lower oesophageal sphincter responses to noxious oesophageal chemical stimuli in the ferret: involvement of tachykinin receptors. AB - Repeated oesophageal acidification is a definitive feature of gastro-oesophageal reflux disease, which in turn is caused by relaxation of the lower oesophageal sphincter (LOS). This study in anaesthetised ferrets investigates the reflex pathways involved in effects of oesophageal acidification on motor function of the LOS, with particular focus on the role of tachykinins. LOS pressure was monitored with a perfused micromanometric sleeve assembly. Oesophageal acidification reduced LOS pressure by 48 +/- 5% until washout with saline. This reduction became larger with repeated tests, and was unaffected in amplitude by acute bilateral vagotomy, although the response became slower in onset. Intra oesophageal capsaicin (0.5% solution) caused a 68 +/- 17% decrease in LOS pressure which remained unchanged with repeated tests. The NK-1 receptor antagonist CP96,345 (1-5 mg/kg intravenous (i.v.) blocked the post-vagotomy LOS responses to both intra-luminal acid and capsaicin. Close intra-arterial (i.a.) injections of capsaicin (1-100 micrograms) gut induced LOS relaxation which was neither vagally nor NK-1 receptor-mediated. Substance P or the selective NK-1 receptor agonist [Sar9, Met(O2)11] substance P (25-500 ng close i.a.) caused a biphasic LOS response, consisting of initial brief contraction followed by prolonged, dose-dependent relaxation. Tetrodotoxin (10 micrograms/kg close i.a.) changed the biphasic response to substance P to excitation only. The neurokinin-1 (NK-1) receptor antagonist CP96,345 (0.3-10 mg/kg i.v.) dose-dependently reduced the inhibitory response to substance P. The excitatory phase of the response to substance P was larger and prolonged after guanethidine (5 mg/kg, i.v.), or propranolol (1 mg/kg, i.v.). L-NAME (100 mg/kg i.v.) reduced the inhibitory phase. The selective NK-2 receptor agonist [beta-Ala8] neurokinin A(4-10) caused LOS excitation only. These data indicate that intra-oesophageal acid causes substance P release from extrinsic afferent nerve endings which activates local inhibitory pathways to the LOS via NK-1 receptors. PMID- 9406125 TI - Distribution of synaptic boutons around identified neurones lying in the cardiac plexus of the guinea-pig. AB - Neurones in the cardiac plexus of the guinea-pig were subdivided into three groups according to their electrophysiological properties and subsequently labelled with neurobiotin. Preparations were counterstained with antibodies to synaptophysin to reveal boutons containing synaptic vesicles. Two of the three groups of cells which, in the electrophysiological studies were found to receive excitatory synaptic inputs were associated with numerous synaptophysin positive boutons. Cells of the other group, which did not appear to receive any synaptic inputs were associated with fewer synaptophysin positive boutons. Wholemount preparations were double stained with antibodies to Protein Gene Product 9.5 and synaptophysin. This revealed most neurones, along with vesicle filled boutons. Ganglia generally contained less than 10 neurones with a range between one and over 30. In wholemount preparations it was found that about 90% of the cells were associated with a large number of synaptophysin positive boutons whilst the remaining cells were associated with very few synaptophysin positive boutons. These results are consistent with the idea that a proportion of cardiac ganglion cells fail to receive a synaptic input and suggest that by staining wholemount preparations of guinea-pig cardiac plexus with antibodies to synaptophysin it is possible to differentiate between groups of cells which receive a synaptic input and those which fail to receive a synaptic input. Approximately 10% of intrinsic cardiac neurones appear to lack a synaptic input. PMID- 9406126 TI - Glutamine and intestinal immune cells in humans. AB - BACKGROUND: Total parenteral nutrition (TPN) is associated with depletion of intestinal immune cells and increased gut permeability (GP). Adding glutamine (GLN) to TPN preserves GP by an unknown mechanism. Intestinal immune cells situated between the enterocytes (intraepithelial lymphocytes, [IEL]) influence GP in vitro. To obtain insight into the underlying mechanism of GLN on GP, we investigated the effects of GLN-supplemented TPN on IEL, immunoglobulin A (IgA) plasma cells and goblet cells, and enterocyte proliferation in intestinal biopsies. METHODS: Twenty patients randomly received GLN-enriched TPN (GT) or isonitrogenous standard TPN (ST). Proliferation and number of immune cells were measured in intestinal biopsies obtained before and after 10 days of TPN. RESULTS: No change in proliferative activity or in number of IgA plasma cells was observed. Goblet cells increased in the ST group, whereas the change seen in the GT group did not reach significance. In the GT group, IEL decreased, whereas in the ST group, no change in the number of IEL was observed. CONCLUSIONS: TPN was not associated with changes in proliferative activity or with depletion of gut immune cells. The data indicate that GLN-supplemented TPN has a different effect on intestinal immune cells compared with standard TPN. PMID- 9406127 TI - Effect of an elemental vs a complex diet on L-citrulline production from L arginine in rat isolated enterocytes. AB - BACKGROUND: L-Arginine and L-glutamine are highly metabolized by intestinal cells, leading to various metabolites, including L-citrulline, which is required for optimal growth. Elemental diets, used in clinical practice to treat growth failure and malnutrition, are very different from complex diets normally consumed. The aim of the present study was to assess the effects of an elemental diet compared with a complex diet on L-arginine metabolism in rat isolated enterocytes and its modulation by L-glutamine. METHODS: Rats were fed the elemental diet (group ED) or the control diet (group C) for 14 days. Villus enterocytes then were isolated, and metabolic capacities or enzyme activities were assessed. RESULTS: The incubation of enterocytes isolated from group C with 0.1 mmol/L L-[U-14C]-arginine led to the production of 125 +/- 25 pmol L citrulline/10(6) cells per 30 minutes. This production showed a twofold increase in the presence of 2 mmol/L L-glutamine. In group ED, L-citrulline synthesis from L-arginine was markedly lower in the absence or in the presence of L-glutamine. This coincided with lower carbamoylphosphate synthase I activity and carbamoylphosphate (CP) content of enterocytes. Other L-arginine and L-glutamine metabolic pathways were not affected. Similar results were obtained when the elemental diet was administered continuously through a gastric catheter or fed by mouth. CONCLUSIONS: L-Glutamine favors the synthesis of L-citrulline from L arginine in isolated enterocytes, probably via an increase in CP production. Changing the diet composition, from a complex to an elemental diet, results in an alteration of the enterocyte capacity to synthesize L-citrulline from L-arginine, irrespective of the rhythm of delivery. PMID- 9406128 TI - Accelerated nitrogen loss after traumatic injury is not attenuated by achievement of energy balance. AB - BACKGROUND: We wanted to determine if achievement of energy balance decreases myofibrillar protein catabolism and nitrogen loss during posttraumatic catabolic illness. METHODS: Surgical intensive care unit of a level I trauma center in a university medical center. Trauma patients expected to be mechanically ventilated for at least 4 days were randomly assigned to one of three parenteral feeding groups: (1) nonprotein calorie group: dextrose and lipid intake equal to measured energy expenditure; (2) total calorie group: dextrose, lipid, and protein intake equal to measured energy expenditure; and (3) hypocaloric group: dextrose and lipid intake equal to 50% of measured energy expenditure. Target protein intake for all groups was 1.7 g/kg body wt. On day 4 of nutrition support, a 24-hour balance study was conducted. Urine urea and total nitrogen production, 3 methylhistidine excretion, energy expenditure, and substrate utilization were measured. RESULTS: Despite significant differences in nonprotein and total calorie balance among the groups, nitrogen loss, nitrogen balance, and catabolic rate were not significantly different. Nitrogen loss correlated with catabolic rate but not with energy expenditure or energy balance. Catabolic rate was associated with energy expenditure but not with energy balance. Nitrogen loss was positively correlated with the percentage of nonprotein energy expenditure met by nonprotein calorie intake. CONCLUSIONS: Achievement of energy balance (nonprotein or total energy) failed to decrease catabolic rate or nitrogen loss acutely in multiple trauma patients. Provision of caloric intake equal to energy expenditure does not seem necessary during the acute phase of posttraumatic catabolic illness. PMID- 9406129 TI - The effects of short-term parenteral nutrition on human liver protein and amino acid metabolism during laparoscopic surgery. AB - BACKGROUND: This study was undertaken to elucidate the specific effects of short term artificial nutrition on human liver protein metabolism. METHODS: Thirty patients undergoing elective laparoscopic cholecystectomy were studied: a control group (n = 16) and a group that received total parenteral nutrition (TPN; n = 14). The nutrition consisted of a balanced i.v. solution of nutrients (17.5 nonprotein kcal/kg body wt, 50% fat, 50% carbohydrates, and 0.1 gN/kg) that was discontinued when the investigation was finished, after a total infusion time of 8.6 +/- 1.0 hours. A liver biopsy specimen was taken as soon as possible after surgery was started, for the determination of the free hepatic amino acid concentrations. In 16 of the patients, L[2H5]phenylalanine was given by i.v. to determine the fractional synthesis rate of total liver protein in a second liver biopsy specimen taken approximately 30 minutes later. RESULTS: The fractional synthesis rate of total liver protein was 15.2% +/- 4.7%/d in the TPN group (n = 7), which was not different from that of the control group (17.7% +/- 3.8%/d, n = 9). However, the free hepatic concentrations of alanine (p < .05) and the essential amino acids increased (p < .001) in the TPN group, whereas the total hepatic amino acid concentrations were comparable between the groups. CONCLUSION: Thus short-term TPN induced specific changes of the free hepatic amino acid concentrations, whereas total liver protein synthesis remained unaffected by the nutrition. PMID- 9406130 TI - Opiate and sedative dependence predicts a poor outcome for patients receiving home parenteral nutrition. AB - BACKGROUND: Home parenteral nutrition (HPN) is used to treat intestinal failure. A minority of HPN patients are dependent on opiates and benzodiazepines to control pain and anxiety. The aim of this study was to determine what effects such drug dependence had on patient outcomes. METHODS: Ten dependent patients were prospectively compared with 10 well-matched, nondependent HPN patients for the same 12-month period. Episodes of line sepsis and other complications were documented and the cost of treatment estimated. Health status was measured using the SF36 and EuroQol instruments. RESULTS: The dependent group had significantly more episodes of central line sepsis (p = .0007) as well as other complications (p = .0002). This led to significantly longer periods of inpatient care (p = .0004) and therefore higher costs of treatment. Health status was lower in the dependent group; they reported more pain (p = .04) and less energy (p = .04). CONCLUSIONS: The complication rate and increased cost of treatment for opiate- and sedative-dependent patients receiving HPN significantly detract from the overall outcome of this therapy. PMID- 9406131 TI - Outcome of cancer patients receiving home parenteral nutrition. Italian Society of Parenteral and Enteral Nutrition (S.I.N.P.E.). AB - BACKGROUND: Indication for home parenteral nutrition (HPN) in cancer patients is controversial because intestinal failure and malnutrition are often only two of the many problems found in such patients that may deserve priority of treatment. METHODS: This was a retrospective study of 75 cancer patients from nine institutions included in the Italian HPN Registry. The patients had a mean weight loss of 12.5%, serum albumin of 3.1 g/dL, lymphocyte count of 1150/mm3, and serum total iron-binding capacity of 190 micrograms/dL. The main indication for HPN was intestinal obstruction (66%); 72% of the patients had metastatic disease. A series of demographic, oncologic, and nutritional characteristics were analyzed in an attempt to predict a possible benefit of HPN. RESULTS: A total of 9897 days of HPN were delivered to 75 cancer patients, for a median of 4 months (range 1 to 15 months) per patient. Sixty-nine patients died while receiving HPN, five had a remission of their intestinal failure, and one chose to stop the treatment. Complications related to parenteral nutrition were as follows: 19 cases of sepsis, 6 catheter occlusions, 4 catheter dislocations, and 2 metabolic imbalances. HPN preserved nutritional status and slightly improved weight, lymphocyte count, serum albumin, and Karnofsky performance status in patients who survived > 3 months. Quality of life during HPN was judged by the clinicians to have improved in only 9% of those who survived < 3 months, but in 68% of the patients who survived for > 3 months. Karnofsky performance status > 50 at the start of HPN was correlated with longer survival (p = .02). CONCLUSIONS: Our study demonstrated a positive effect of HPN on nutritional status and quality of life in patients who survived > 3 months and suggests that HPN should be avoided when Karnofsky performance status is < 50. PMID- 9406132 TI - Comparison of bone marrow toxicity of medium-chain and long-chain triglyceride emulsions: an in vitro study in humans. AB - BACKGROUND: In this study, we evaluated the in vitro bone marrow toxicity of two lipid emulsions containing either long-chain triglycerides (LCT) or a mixture of medium-chain triglycerides (MCT) and LCT. METHODS: Bone marrow cells were obtained from six healthy subjects and were cultured for 14 days after a 24-hour preincubation with various concentrations (from 0 to 10 mg/mL) of LCT- and LCT/MCT-based lipid emulsions. RESULTS: Compared with controls (no preincubation with lipid emulsion), both lipid emulsions significantly inhibited by 50% to 70% colony formation of all the human bone marrow cells cultured from a triglyceride concentration of 0.5 mg/mL (p < .05). Erythroid burst-forming unit (BFU-E) formation was significantly more inhibited with LCT/MCT emulsion than with LCT emulsion (p < .05). The inhibition of granulocyte-macrophage colony-forming unit (GM-CFU) and mixed granulocyte-erythrocyte-monocyte-megakaryocyte colony-forming unit (GEMM-CFU) formation did not significantly differ with the two emulsions. CONCLUSIONS: Both LCT- and LCT/MCT-based lipid emulsions strongly inhibit colony formation by human bone marrow cells. BFU-E colony formation is more sensitive to LCT/MCT inhibition than to LCT. PMID- 9406133 TI - Insulin resistance and the alterations of glucose transporter-4 in adipose cells from cachectic tumor-bearing rats. AB - BACKGROUND: Insulin resistance may play an important role in cancer cachexia; however, its mechanisms remain to be clarified. METHODS: Cellular mechanisms of insulin resistance in tumor-bearing rats (TBR) were investigated in isolated adipose cells by measuring 3-O-[14C]methyl glucose transport activity and glucose transporter-4 (GLUT4) protein in low-density microsomes at a basal state and in the plasma membrane at an insulin-stimulated state. RESULTS: The insulin stimulated glucose transport activity in adipose cells from TBR was significantly lower than that of control rats (CTR) (0.51 +/- 0.25 and 2.27 +/- 0.11 fmol/cell/min, respectively). The amount of GLUT4 in low-density microsomes at a basal state and in plasma membrane at an insulin-stimulated state was less in TBR than in CTR. CONCLUSIONS: These data suggest that the insulin resistance seen in the adipose cells of these tumor-bearing rats was caused in part by both a decreased amount of GLUT4 protein in a basal state and a decreased translocation of GLUT4 in response to insulin stimulation. PMID- 9406134 TI - Stimulation of gallbladder emptying by intravenous lipids. AB - BACKGROUND: Few studies have dealt with the effect of i.v. administration of lipids on gallbladder emptying, and the results have been conflicting. METHODS: Five healthy volunteers, three women and two men, aged 26 to 54 years, (mean, 29 years) were studied. Gallbladder emptying was assessed by means of real-time ultrasonography. RESULTS: In all subjects, the infusion of a 10% fat emulsion (Intralipid; Kabivitrum, Stockholm) over 3 hours caused a reduction in gallbladder volume. This effect was statistically significant at about 80 minutes of lipid infusion and became progressively more marked as the infusion progressed, reaching a reduction of approximately 30% during the third hour of infusion. A significant relationship (p < .001) was found between the concentration of serum triglycerides and the degree of gallbladder volume decrease. In control studies, infusion of physiologic saline containing glycerol, the excipient of intralipid, caused no significant changes in gallbladder volume. CONCLUSIONS: The results indicate that i.v. infusion of lipids is able to stimulate significantly contraction of human gallbladder. PMID- 9406135 TI - Prophylactic locking of enteral feeding tubes with pancreatic enzymes. AB - BACKGROUND: Obstruction of feeding tubes is a common mechanical complication associated with enteral feeding. Standard methods of flushing are not always effective. METHODS: This study was conducted with patients receiving enteral feeding via nasogastric, nasoenteral, gastrostomy, or jejunostomy tubes to determine if prophylactic use of pancreatic enzymes would maintain patency of feeding tubes. Interrupted feeding regimens were used. Control patients (n = 24) received only water for flushing. After water flushing of the tube, study patients (n = 33) received a 5-mL suspension of pancreatic enzyme containing the following enzyme activity (in Federation Internationale Pharmaceutique [FIP] units): lipase, 2000; amylase, 1500; and protease, 100. The suspension also contained 90 mg of NaHCO3 to maintain a pH of 7.5. The mean duration of observation in the control and study groups was 25 and 48 days, respectively. RESULTS: Compared with eight episodes (23.5%) of tube occlusions in the control group (n = 34), there was only one episode (2.6%) in the study group (n = 38). This difference was significant by the test of proportions (z = 2.68, p = .01). CONCLUSIONS: In addition to routine water flushing, the routine prophylactic use of pancreatic enzyme-sodium bicarbonate suspension (pH 7.5) prevents occlusion of feeding tubes. PMID- 9406136 TI - Role of intestinal bacteria in nutrient metabolism. AB - The human large intestine contains a microbiota, the components of which are generically complex and metabolically diverse. Its primary function is to salvage energy from carbohydrate not digested in the upper gut. This is achieved through fermentation and absorption of the major products, short chain fatty acids (SCFA), which represent 40-50% of the available energy of the carbohydrate. The principal SCFA, acetate, propionate and butyrate, are metabolized by the colonic epithelium (butyrate), liver (propionate) and muscle (acetate). Intestinal bacteria also have a role in the synthesis of vitamins B and K and the metabolism of bile acids, other sterols and xenobiotics. The colonic microflora are also responsive to diet. In the presence of fermentable carbohydrate substrates such as non-starch polysaccharides, resistant starch and oligosaccharides, bacteria grow and actively synthesize protein. The amount of protein synthesis and turnover within the large intestine is difficult to determine, but around 15 g biomass is excreted in faeces each day containing 1 g bacterial-N. Whether bacterially synthesized amino acids are ever absorbed from the colon remains unclear. Finally, individual colonic micro-organisms such as sulphate-reducing bacteria, bifidobacteria and clostridia, respond selectively to specific dietary components in a way that may be important to health. PMID- 9406137 TI - Hypoenergetic nutrition support in hospitalized obese patients: a simplified method for clinical application. PMID- 9406138 TI - Analysis of responses to human synthetic adrenomedullin and calcitonin gene related peptides in the hindlimb vascular bed of the cat. AB - Vasodilator responses to human adrenomedullin (hADM), a newly discovered hypotensive peptide, human calcitonin gene-related peptide-alpha (hCGRP-alpha) and hCGRP-beta, which share structural homology with hADM, were compared in the hindlimb vascular bed of the cat under constant flow conditions. Injections of hADM (0.003-1 nmol), hCGRP-alpha, and hCGRP-beta (0.003-0.3 nmol) into the perfusion circuit caused dose-related decreases in hindlimb perfusion pressure. Vasodilator responses to hCGRP-alpha and hCGRP-beta were similar in potency and duration, and the doses of hCGRP-alpha and hCGRP-beta required to reduce hindlimb perfusion pressure 40 mm Hg (ED40 mm Hg) were significantly lower than the ED40 mm Hg for hADM. The duration of the hindlimb vasodilator responses to hCGRP-alpha and hCGRP-beta were significantly longer than the duration of the response to hADM. Amylin, a peptide that shares structural homology with ADM and with CGRP, had no significant effect on hindlimb perfusion pressure when injected in doses up to 1 nmol. Decreases in hindlimb perfusion pressure in response to hADM, hCGRP alpha, and hCGRP-beta were not altered by L-N5-(1-iminoethyl)-ornithine (L-NIO) in a dose of the nitric oxide synthase inhibitor that decreased the vasodilator response to acetylcholine or by the cyclooxygenase inhibitor, meclofenamate, in a dose that decreased the vasodilator response to archidonic acid. The present data demonstrate that hADM, hCGRP-alpha, and hCGRP-beta have potent, but relatively short-lasting, vasodilator activity, and that vasodilator responses are not dependent on the release of nitric oxide or vasodilator prostaglandins in the hindlimb vascular bed of the cat. PMID- 9406139 TI - Effect of short-term treatment with a monoclonal antibody to P-selectin on balloon catheter-induced: intimal hyperplasia, re-endothelialization, and attenuation of endothelial-dependent relaxation. AB - The effects of an anti-P-selectin monoclonal antibody (MAb, PB1.3; Cytel Corporation) on neoendothelialization; neoendothelial function, as evidenced by acetylcholine-induced relaxation (nitric oxide formation); and intimal hyperplasia following embolectomy catheter-induced injury to the rabbit thoracic aorta were investigated. Catheter injury was induced in two groups of New Zealand White rabbits. One group received no treatment, while the second group received short-term treatment with the MAb (i.p., immediately before and 12 h after induction of catheter injury). A third group underwent a sham operation and served as uninjured controls. Following sacrifice at 2 weeks after injury, aortic rings were assessed for degree of intimal hyperplasia, neoendothelial morphology (scanning electron microscopy), and acetylcholine-induced relaxation. Aortic tissue from catheter-injured animals that received treatment exhibited improved neoendothelial morphology, as compared with tissue from untreated but catheterized animals; however, no statistically significant attenuation of the hyperplastic response or improvement in the attenuated neoendothelial-dependent acetylcholine-induced relaxant response that is characteristic of neoendothelium that forms after catheter denudation was observed. These data suggest that short term attenuation of P-selectin-mediated polymorphonuclear leukocyte (PMN)/endothelium, PMN/platelet interactions, and/or thrombin formation beneficially affects neoendothelialization of the vascular wall following balloon catheter-induced injury. PMID- 9406141 TI - Endothelin contraction in pig coronary artery: receptor types and Ca(2+) mobilization. AB - Endothelin is one of the most potent vasoconstrictors known. It plays an important role in the regulation of vascular tone and in the development of many cardiovascular diseases. This study focuses on the receptor types and the Ca2+ mobilization responsible for endothelin-1 (ET-1) contraction in de endothelialized pig coronary artery rings. ET-1 contracted the artery rings with an EC50 = 6.5 +/- 1 nM and a maximum contraction which was 98.6 +/- 9% of the contraction produced by 60 mM KCl. BQ123 (5 microM), an ETA antagonist, reversed 78 +/- 3% of the ET-1 contraction (50 nM). IRL1620, a selective ETB agonist, produced 23 +/- 3% of the total ET-1 contraction with an EC50 = 12.7 +/- 2 nM. More than 85% of the contraction due to 100 nM IRL 1620 was inhibited by 200 nMBQ788, an ETB antagonist. Therefore, approximately 80% of the ET-1 contraction in this artery occurred via ETA receptors, and the other 20% was mediated by ETB receptors. To assess the Ca2+ pools utilized during the ET-1 response, ET-1 contraction was also examined in medium containing an L-type Ca2+ channel blocker nitrendipine, and in Ca2+ free medium containing 0.2 mM EGTA. In Ca2+ containing medium the contraction elicited by ET-1 was 98.6 +/- 9% of the KCl contraction, however, in the presence 10 microM nitrendipine the ET-1 induced contraction was 54 +/- 7% of the KCl contraction, and in Ca(2+)-free medium it was 13 +/- 2%. Similarly, the IRL 1620 contractions in Ca2+ containing medium, in the presence of nitrendipine and in Ca(2+)-free medium were 22.4 +/- 3%, 12 +/- 3% and 11 +/- 2% of the KCl response respectively. Thus, both ETA and ETB contractions utilize extracellular Ca2+ pools via L-type Ca2+ channels and other undefined route(s), as well as intracellular Ca2+ pools. In the pig coronary artery smooth muscle, ET 1 contractions occur predominantly via ETA receptors, with ETB receptors using similar Ca2+ mobilization pathways, but the ETB receptors appear to use the intracellular Ca2+ stores to a greater extent. PMID- 9406140 TI - Differential regulation of G-protein expression by vasoactive peptides. AB - Vasoactive peptides such as angiotensin II (AII), atrial natriuretic peptide (ANP) and vasopressin play an important role in the regulation of blood pressure. We have recently shown an augmentation of Gi alpha levels in heart and aorta from genetic and experimentally-induced hypertensive rats, which may be attributed to the increased levels of vasoactive peptides. We have therefore investigated the effect of AII and ANP on the expression of G-proteins (Gi alpha and Gs alpha) in cultured vascular smooth muscle cells (VSMC) and their relationship with adenylyl cyclase activity. Exposure of VSMC with AII resulted in the augmentation of the levels of Gi alpha-2 and Gi alpha-3 proteins and Gi alpha-2 and Gi alpha-3 mRNA and not of Gs alpha as determined by immunoblotting and Northern blotting techniques respectively. However, the stimulatory effects of N-ethylcarboxamide adenosine (NECA) and isoproterenol on adenylyl cyclase was diminished by AII treatment, whereas the inhibitory effects of AII and C-ANP4-23 were completely attenuated. On the other hand, pretreatment of the cells with C-ANP4-23 resulted in the reduction of the levels of Gi alpha-2 and Gi alpha-3 and not of Gs alpha. The inhibitory responses of adenylyl cyclase to C-ANP4-23 and AII were also attenuated and the stimulatory effects of GTP gamma S and other agonists were significantly augmented. These data indicate that AII and ANP modulate the expression of Gia protein in a different manner. It may be suggested that the enhanced levels of Gi alpha protein observed in hypertension may be attributed to the augmented levels of AII and not to ANP. PMID- 9406142 TI - Activation of the neutrophil and loss of plasma glutathione during Mg-deficiency- modulation by nitric oxide synthase inhibition. AB - Sprague-Dawley rats (200 g) were fed either a Mg-deficient or Mg-sufficient diet for 3 weeks. An enriched neutrophil fraction (> 85%) was isolated from the blood by sodium metrizoate/dextran gradient centrifugation. Using the superoxide dismutase. (SOD)-inhibitable cytochrome c reduction assay, the basal activity of neutrophils isolated from the Mg-deficient rats were found elevated 5 fold after two weeks, and up to approximately 7 fold after three weeks on the diet. Upon challenge by phorbol myristate acetate (PMA), unlike the Mg-sufficient cells, the Mg-deficient cells exhibited no significant activation. Treatment of the Mg deficient rats with the nitric oxide (NO)-synthase inhibitor, NG-nitro-L-arginine methyl ester (L-NAME) in the drinking water, significantly attenuated the basal superoxide producing activity of the neutrophils and partially restored its response to PMA challenge. In association with the neutrophil activation. Mg deficiency resulted in 70% decrease in plasma glutathione and 220% increase in Fe promoted, thiobarbituric acid reactive substance (TBARS) levels; both changes were significantly attenuated by L-NAME treatment. The results suggest that neutrophils from Mg-deficient rats are activated endogenously to generate oxy radicals which might directly mediate the in vivo peroxidative indices during Mg deficiency. Furthermore, the neutrophil activity was lowered by NO-synthase inhibition suggesting that NO overproduction during Mg-deficiency participates in the neutrophil activation process. PMID- 9406143 TI - Velocity of translation of single actin filaments (AF) by myosin heads from antigen-sensitized airway smooth muscle. AB - We have previously reported increased velocity of shortening (Vo) in the sensitized airway (0.36 1o/s, +/- SE) smooth muscle compared to the control (0.26 1o/s, +/- 0.017 SE) and subsequent experiments indicated this was due to increased phosphorylation of the 20 kDa myosin light chain resulting from increased total myosin light chain kinase activity. The motility assay technique described by Kron and Spudich was employed to determine whether additionally the molecular motor (actomyosin crossbridge) itself was altered in airway smooth muscle by ragweed pollen sensitization. The motility assay measures the velocity of actin filament translation by myosin molecules. The negative results of the motility assay were valuable in determining that the pathogenesis of allergic bronchospasm is not at contractile protein level but at regulatory enzyme level. PMID- 9406144 TI - Smooth muscle contractility and protein tyrosine phosphorylation. AB - During the last 5 years several studies have documented an involvement of protein tyrosine kinases (PTKs) in smooth muscle contraction and Ca2+ mobilization. Most of these studies have utilized highly selective inhibitors of PTKs, genistein and tyrphostin and have shown that these inhibitors attenuated smooth muscle contraction induced by growth factors-epidermal growth factor (EGF) and platelet derived growth factor (PDGF) and several vasoactive peptides. It has also been demonstrated that inhibitors of protein tyrosine phosphatases (PTPases) such as vanadate and pervanadate mimic growth factors and vasoactive peptides in causing the contraction of smooth muscle. In this brief review, we have summarized some of the recent observations suggesting a possible link between protein tyrosine phosphorylation pathway and smooth muscle contraction. PMID- 9406145 TI - Coronary artery smooth muscle in culture: migration of heterogeneous cell populations from vessel wall. AB - A method for establishing primary cultures of smooth muscle cells (SMCs) from the porcine coronary artery without either microdissection and/or enzymatic dispersion was developed using selective migration of cells from coronary explants in vitro. This culture method relies on the heterogeneity of cell types and differences in their migration and adherence ability to separate SMC from contaminating fibroblasts or endothelial cells. The cell type was determined by immunohistochemical staining with monoclonal antibodies to SM alpha-actin, SM myosin, h-caldesmon and von Willebrand factor. The first wave of migration (1-7 days) consisted of a mixture of fibroblasts and SMCs. Only SMCs were present in the second wave of migration (7-14 days). Endothelial cells, which exhibited a lower capacity for migration and adherence, were restricted to the third wave of migration (14-21 days). Cells obtained from the second wave of migration exhibited the characteristic single-layered, aligned, hill-and-valley pattern of SMCs when confluent. Quiescence was attained 4-5 days after removal of serum, as established by [3H]-thymidine incorporation. Stimulation of the quiescent SMCs with 20% FBS resulted in a synchronous re-entry into the cell-cycle with S phase reached 15-18 h later. The SMCs prepared using this protocol thus exhibit the structural markers and capacity to undergo phenotypic modulation that are characteristic of SMCs in vivo. This approach to establishing primary cultures of SMCs offers the advantage of selecting for the subpopulation of cells capable of migration in response to injury or growth factor stimulation. PMID- 9406146 TI - An overview of the influence of ACE inhibitors on fetal-placental circulation and perinatal development. AB - The renin-angiotensin system is associated with a variety of pathophysiological processes in many organ systems, and is known to be involved in the normal regulation of blood pressure and in the pathogenesis of renovascular hypertension. Angiotensin II is a multifunctional hormone that manifests its properties by interacting with two major subtypes of cell surface receptors (AT1 and AT2). Angiotensin converting enzyme (ACE) inhibitors are able to modify the actions of the renin-angiotensin system, and are indicated for the treatment of hypertension and heart disease. The antihypertensive effects of ACE inhibiting drugs are related to their ability to block the conversion of the decapeptide, angiotensin I, to the potent pressor octapeptide, angiotensin II. ACE inhibitors have been implicated in fetopathies in humans and perinatal mortality in rats, rabbits, sheep and baboons. Human fetopathies were seen when ACE inhibitors were given around the 26th week of gestation. The major adverse effects in babies include: oligohydramnios, renal tubular dysgenesis, neonatal anuria, calvarial and pulmonary hypoplasia, mild to severe intrauterine growth retardation, persistent patent ductus arteriosus and fetal or neonatal death. These developmental anomalies are thought to be partly due to a direct action of ACE inhibitors on the fetal renin-angiotensin system and partly due to the ischemia resulting from maternal hypotension and decreases in fetal-placental blood flow and oxygen/nutrient delivery to the fetus. The purpose of this review is to briefly discuss the pathophysiological role of the renin-angiotensin system, the therapeutic uses of ACE inhibitors in pregnant patients and to focus primarily on the major fetotoxic effects of ACE inhibitors encountered in humans and animal models. I will also review our recent data which show that capozide (captopril + hydrochlorothiazide) not only produces oligohydramnios but also disturbs the balance of glucose and NaCl in the maternal plasma and amniotic fluid of the rat. PMID- 9406148 TI - Reversal of phosphate induced decreases in force by the benzimidazole pyridazinone, UD-CG 212 CL, in myofilaments from human ventricle. AB - UD-CG 212 Cl, (Fig. 1: 4,5-dihydro-6-[2-(4-hydroxyphenyl)-1 H-benzimidazole-5-yl] 5-methyl-3(2H)-pyridazinone), is the primary metabolite of the positive inotropic agent pimobendan (UDCG 115 BS, Acardi). Our previous studies showed in detergent extracted preparations of canine ventricular muscle that sub-nanomolar concentrations of UD-CG 212 Cl increased submaximal myofilament force, but only when the activation state had been altered by relatively high (5-10 mM) concentrations of inorganic phosphate (Pi) or relatively low (20 microM) concentrations of MgATP. In the present study, we investigated the effects of UD CG 212 Cl on the pCa-force relationship of detergent extracted bundles of human cardiac fibers before and after addition of Pi. As expected, treatment with 5 mM Pi depressed maximal force at pCa 4.5 by 27.0 +/- 0.4% (mean +/- SEM). Force generated at the half-maximally activating Ca2+ concentration (pCa50) of control fibers (5.98 +/- 0.2) was significantly (p < .05) reduced following the addition of 5 mM Pi (pCa50 = 5.69 +/- 0.3). The addition of UD-CG 212 Cl over a range of concentrations (10(-11)-10(-6) M) had no effect on Ca(2+)-sensitivity under control conditions, but in the presence of 5 mM Pi, there was a 23.1 +/- 0.1% increase in the percent maximal force at pCa5.9. Ca(2+)-sensitivity was also significantly increased in the presence of Pi and 10(-8) M UD-CG 212 Cl (pCa50 = 5.74 +/- 0.3, p < .05). We conclude that UD-CG 212 Cl potentially increases sub maximal force of human ventricular myofilaments with an inotropic action depending on a state of myofilament activation associated with ischemic conditions. PMID- 9406147 TI - Differential nature of cross-talk among three G-coupled receptors regulating adenylyl cyclase in rat cardiomyocytes chronically exposed to receptor agonists. AB - Chronic exposure of cells to cognate agonists has been established to cause homologous desensitization of G protein-coupled receptors. In this work, we show that exposure of adult rat cardiomyocytes to isoproterenol (ISO) for 24 h led to the desensitization of beta-adrenoceptor (beta-AR) coupled adenylyl cyclase (AC) activity, which was associated with an increased inhibition of AC by M2 muscarinic receptor (MR) agonist, carbachol (Cch), and a decreased inhibition of AC by A1-adenosine receptor (AdR) agonist, N6-phenylisopropyladenosine (R-PIA). Chronic exposure of cells to Cch caused the desensitization of M2-MR-coupled AC, decreased the inhibitory action of R-PIA on AC and increased ISO-stimulated AC, while chronic exposure to R-PIA caused the desensitization of A1-AdR-coupled AC and modestly increased ISO-stimulated AC without any significant effect on Cch inhibition of the enzyme. Thus, chronic exposure of cardiomyocytes revealed for the first time a more complex and differential nature of cross-talk among the three major G-coupled receptors in modulating AC. PMID- 9406149 TI - Expression of fibroblast growth factor receptor-1 in rat heart H9c2 myoblasts increases cell proliferation. AB - Basic fibroblast growth factor (FGF-2) plays an important role in myocardial growth and development and in particular cardiac myocyte proliferation. FGF-2 exerts its effects by binding to cell surface receptors (FGFR-1) of the tyrosine kinase family. We have detected the presence of both long and short isoforms of FGFR-1 in embryonic and adult mouse heart. In this report, we have examined the ability of long and short FGFR-1 isoforms to signal a mitogenic response. Assessment of RNA from rat myoblast H9c2 cells by reverse transcriptase polymerase chain reaction and RNA blotting revealed that they were deficient in transcripts corresponding to long and short FGFR-1 species. Hybrid genes containing the cDNAs coding for long and short FGFR-1 isoforms directed by the myosin light chain-2 promoter and simian virus 40 enhancer sequences, were used to transiently transfect H9c2 cells. Total tyrosine phosphorylation was increased 2.0 and 2.6 fold in H9c2 cells transfected with the long and short FGFR-1 isoforms, respectively, compared to 'control' transfected H9c2 cells. This was accompanied by a 2.1 and 2.0 fold increase in DNA synthesis, as measured by tritiated thymidine incorporation, in H9c2 cells expressing the long and short FGFR-1 isoforms, respectively. To assess effects on proliferation, H9c2 cells were stably transfected with the myosin light chain-2/FGFR-1 cDNA genes. The rate of proliferation was increased 1.6 and 3.1 fold in H9c2 cells stably expressing the long and short FGFR-1 isoforms, respectively, compared to 'control' H9c2 cells. In contrast to non transfected H9c2 cells, treatment of H9c2 cells stably expressing long FGFR-1 with FGF-2 for 24 h resulted in a slight increase (1.3 fold, p < 0.02) in cell number. However, a greater response (1.5 fold, p < 0.0005) was observed with H9c2 cells stably expressing short FGFR-1 after treatment with FGF-2. These results suggest that both long and short FGFR-1 isoforms are capable of signalling a mitogenic response. PMID- 9406150 TI - Identification of an HMG-like protein involved in regulation of Na+/H+ exchanger expression. AB - In this study we characterized regulation of the Na+/H+ exchanger promoter in several tissue types. A conserved poly (dA:dT) region was important in regulation of the promoter. Nuclear extracts from rat myocardium and from mouse proximal tubule cells protected the poly (dA:dT) region of the NHE1 promoter. A protein from nuclear extracts also bound to the poly (dA:dT) element in gel mobility shift binding assays. The binding was specific and was removed by mutations in the poly (dA:dT) region. Characterization of the binding to the poly (dA:dT) region in gel mobility shift assays showed that it was reduced by high concentrations of the divalent cations Mg++ and Mn++. The inhibition by divalent cations was reduced by decreasing the pH of the binding assay. N-terminal sequencing of the poly (dA:dT) binding protein showed that it was a member of the HMG (high mobility group) family of nuclear proteins which are important in cell growth and proliferation. The results are the first direct detection of a protein that regulates the NHE1 promoter. PMID- 9406151 TI - Cytochemical and immunocytochemical localization of Na,K-ATPase alpha subunit isoenzymes in the rat heart. AB - In order to understand the functional significance of Na,K-ATPase subunits as well as their isoenzymes, a precise subcellular localization of these in the myocyte is a crucial prerequisite. Cytochemical, immunofluorescence, preembedding immunogold and horse radish peroxidase-diaminobenzidine methods, demonstrated alpha 1 isoenzyme immunoreactivity on the sarcolemma, T-tubules and the subsarcolemmal cisterns of the adult cardiac myocytes. Cytochemically, ouabain resistant Na,K-ATPase precipitate was localized only in the subsarcolemmal cisterns and junctional sarcoplasmic reticulum. For alpha 2 isoenzyme, immuno reactivity was demonstrated on the sarcolemma as well as in all areas of the myocytes in particularly a close proximation to the sarcoplasmic reticulum and microsomes. For alpha 3 isoenzyme, only a weak insignificant signal was noted on the sarcolemma, intercalated disc and sarcoplasm. It is suggested that cytochemical ouabain resistant precipitate present in subsarcolemmal cisterns and junctional sarcoplasmic reticulum represent alpha 1 isoenzyme of Na,K-ATPase. A differential as well as unique localization of alpha subunit isoenzymes of Na,K ATPase in specific structures of cardiac myocytes may suggest importance in physiological function at these sites. PMID- 9406152 TI - Estradiol modulates the sodium pump in the heart sarcolemma. AB - Cardiovascular effects of estrogens and particularly that of estradiol involve protection of the heart against ischemia. These effects were believed to be mainly indirect, mediated via changes in the blood and blood vessels. In the present paper a direct action of estradiol on the heart is demonstrated. Estradiol stimulates (p < 0.001) the Na,K-ATPase activity of cardiac sarcolemmal membranes by stimulating in an allosteric manner, the activation of the enzyme by potassium. The latter activation involves also an increase in affinity to potassium of the potassium binding sites on the enzyme molecule, but remains without any effect on the capacity and KD value of specific ouabain binding to the Na,K-ATPase. Estradiol is also antagonizing the depression of Na,K-ATPase activity that may be caused by ischemia and it is stimulating (p < 0.01) the ouabain-sensitive uptake of 86Rb into the heart cells. Our results indicate, that in addition to the known indirect effects of estradiol on the heart, the hormone also stimulates the activity and improves the kinetics of interaction of cardiac sarcolemmal Na,K-ATPase with ATP as well as with Na+ and K+ ions. This direct action may also account for the cardioprotective effects of estradiol. PMID- 9406153 TI - Thyroid hormones differentially affect sarcoplasmic reticulum function in rat atria and ventricles. AB - The present study was undertaken to compare the effects of hypothyroidism and hyperthyroidism on sarcoplasmic reticulum (SR) Ca(2+)-pump activity, together with assessment of the functional role of SR in providing activator Ca2+ under these altered thyroid states. In response to a shift from hypothyroid to hyperthyroid state, a 10 fold and 2 fold increase in SR Ca(2+)-pump activity in atria and ventricles, respectively, were observed. This was associated with the 8 9 fold increases in atrial contractility (+dT/dt) and relaxation (-dT/dt), but only with a 3-4 fold increase in their ventricular counterparts. Also, the recirculation fraction of activator Ca2+ (RFA) increased to a far greater extent in atria (4 fold) than in papillary muscles, and the relative increment in inhibition of developed tension by ryanodine became 3 times larger in atria than in papillary muscles. A positive force-frequency relationship (FFR) was observed in hypothyroid atria, whereas the hyperthyroid atria, hypothyroid and hyperthyroid papillary muscles showed a negative FFR. These results suggest the greater role of transsarcolemmal (SL) Ca2+ and smaller role of SR Ca2+ in activating contraction in hypothyroid atria compared to other preparations. Thyroid hormones decrease the contribution of SL and increase that of SR in providing activator Ca2+ to the greater extent in atria than in ventricles. This effect of thyroid hormones is based on larger stimulation of SR Ca(2+)-pump in atria compared to ventricles. PMID- 9406154 TI - Dietary and physiological studies to investigate the relationship between calcium and magnesium signalling in the mammalian myocardium. AB - This study employs both dietary and physiological studies to investigate the relationship between calcium (Ca2+) and magnesium (Mg2+) signalling in the mammalian myocardium. Rats maintained on a low Mg2+ diet (LMD; 39 mg Kg-1 Mg2+ in food) consumed less food and grew more slowly than control rats fed on a control Mg2+ diet (CMD; 500 mg Kg-1 Mg2+ in food). The Mg2+ contents of the heart and plasma were 85 +/- 3% and 34 +/- 6.5%, respectively relative to the control group. In contrast, Ca2+ contents in the heart and plasma were 177 +/- 5% and 95 +/- 3%. The levels of potassium (K+) was raised in the plasma (129 +/- 16%) and slightly decreased in the heart (88 +/- 6%) compared to CMD. Similarly, sodium (Na+) contents were slightly higher in the heart and lowered in the plasma of low Mg2+ diet rats compared to control Mg2+ diet rat. Perfusion of the isolated Langendorff's rat heart with a physiological salt solution containing low concentrations (0-0.6 mM) of extracellular magnesium [Mg2+]o resulted in a small transient increase in the amplitude of contraction compared to control [Mg2+]o (1.2 mM). In contrast, elevated [Mg2+]o (2-7.2 mM) caused a marked and progressive decrease in contractile force compared to control. In isolated ventricular myocytes the L-type Ca2+ current (ICa,L) was significantly (p < 0.001) attenuated in cells dialysed with 7.1 mM Mg2+ compared to cells dialysed with 2.9 microM Mg2+. The results indicate that hypomagnesemia is associated with decreased levels of Mg2+ and elevated levels of Ca2+ in the heart and moreover, internal Mg2+ is able to modulate the Ca2+ current through the L-type Ca2+ channel which in turn may be involved with the regulation of contractile force in the heart. PMID- 9406155 TI - Biological significance of phosphorylation and myristoylation in the regulation of cardiac muscle proteins. AB - Post-translational modification has long been recognized as a way in which the properties of proteins may be subtly altered after synthesis of the polypeptide chain is complete. Amongst the moieties most commonly encountered covalently attached to proteins are oligosaccharides, phosphate, acetyl, formyl and nucleosides. Protein phosphorylation and dephosphorylation is one of the most prevalent and best understood modifications employed in cellular regulation. The bovine heart calmodulin-dependent cyclic nucleotide phosphodiesterase (CaMPEDE) can be phosphorylated by cAMP-dependent protein kinase, resulting in a decrease in the enzyme's affinity for Ca2+ and calmodulin (CaM). The phosphorylation of CaMPDE is blocked by Ca2+ and CaM and reversed by the CaM-dependent phosphatase (calcineurin). The dephosphorylation is accompanied by an increase in the affinity of the phosphodiesterase for CaM. Analysis of the complex regulatory properties of CaMPDE has led to the suggestion that fluxes of cAMP and Ca2+ during cell activations are closely coupled and that the CaMPDE play a key role in the signal coupling phenomenon. The high molecular weight calmodulin binding protein (HMWCaMBP) was phosphorylated by cAMP-dependent protein kinase. Phosphorylation of HMWCBP was higher in the absence of Ca2+/CaM then in the presence of Ca2+/CaM and reversed by the CaM-dependent phosphatase. Recently, it has become apparent that the binding of myristate to proteins is also widespread in eukaryotic cells and viruses and certainly is of great importance to the correct functioning of an organism. Myristoyl CoA:protein N-myristoyltransferase (NMT) catalyses the attachment of myristate to the amino-terminal glycine residue of various signal transduction proteins. Cardiac tissue express high levels of cAMP-dependent protein kinase whose catalytic subunit is myristoylated. The subcellular localization of bovine cardiac muscle NMT indicated a majority of the activity was localized in cytoplasm. Under native conditions the enzyme exhibited an apparent molecular mass of 50 kDa. Recovery of NMT activity, from both cytosol and particulate fractions, was found to be higher than the total activity in crude homogenates, suggesting that particulate fraction may contain an inhibitory activity towards NMT. Research in our laboratory has been focusing on the covalent modification of proteins and regulation of various signal transduction proteins. This special review is designed to summarize some aspects of the current work on co- and post-translational modification of proteins in cardiac muscle. PMID- 9406156 TI - Contributions of increased efficiency and capacity of protein synthesis to rapid cardiac growth. AB - Rapid cardiac growth depends upon faster synthesis than degradation of protein. The rate of protein synthesis is determined by the efficiency with which the existing components of the ribosome cycle make protein and by the quantity of the components that are present. The tissue content of RNA is taken as an index of the capacity of synthesis and efficiency is expressed as the amount of protein formed per amount of RNA over a certain time period. The efficiency of synthesis is regulated by hormones, including insulin, agents that increase cAMP, alpha adrenergic agonists, endothelin I and angiotensin II. In addition, provision of non-carbohydrate substrates and mechanical factors such as stretch and contraction increase efficiency. Impaired energy availability as occurs in anoxic or ischemic muscle decreases efficiency. Increased phosphorylation of ribosomal protein, S6, or of the peptide chain initiation factor, elF-4E, have been suggested as mechanisms to regulate efficiency of mRNA translation. Increased efficiency of synthesis accounts for cardiac growth in the first few days following aortic banding, pulmonary artery constriction and thyroxine administration. Decreased efficiency accounts for cardiac atrophy in heterotopic transplanted hearts during the first 3 days following transplantation. The capacity of synthesis is increased by insulin, thyroid hormone, activators of protein kinase C, agents that increase cAMP, and endothelin-1. Stretch of the ventricular wall and contraction of cultured neonatal myocytes accelerates ribosome formation. An increased rate of ribosomal DNA transcription accounts for accelerated ribosome formation and depends on increased activity of a transcription factor, upstream binding factor (UBF). The activity of UBF is increased either by increased rates of synthesis or by phosphorylation of the protein. Increased capacity of synthesis is a major contributor to rapid cardiac growth in the newborn heart and after several days of pressure overload. PMID- 9406158 TI - An endogenous positive inotropic factor (EPIF) from porcine heart: its effects on sarcoplasmic reticular (SR) Ca2+ metabolism. AB - We have isolated an endogenous positive inotropic factor (EPIF) from porcine left heart ventricular tissue, which demonstrated to have only weak digitalis-like properties including the inhibition of myocardial Na+,K(+)-ATPase. EPIF completely lacks digitalis-like toxicity such as after-contractions in larger doses. In our recent studies, we have demonstrated that EPIF produces a decrease in the amplitude of the post-rest rapid cooling contracture which indicated that EPIF may release Ca2+ from the sarcoplasmic reticulum. In the present study, the effects of EPIF were investigated on the Ca2+ uptake and release properties of SR enriched membrane vesicles from rat heart. At pH 6.8 and in the presence of oxalate, EPIF dose-dependently inhibited the ATP-dependent uptake of Ca2+ by SR vesicles. Concentrations as low as 25 ul (in 1 mL uptake medium) of EPIF caused a 45-47% reduction in the uptake of Ca2+ within 3-4 min. Increases in EPIF concentration to 50 ul/mL caused additional reduction of only 15-20% in the uptake of Ca2+. Concentrations of 25 ul/mL of EPIF had little or no effects on passive release of actively loaded Ca2+ in SR vesicles. On doubling the concentrations to 50 ul/mL EPIF, however, enhanced the release of Ca2+ by 25-28% during 1-2 min and 44-48% after 4 min of incubation of Ca2+ loaded vesicles in the release medium. Relatively smaller effects of EPIF on Ca2+ release implies that EPIF may mainly lower the uptake of Ca2+ in SR. This reduced uptake of Ca2+ may be explained by the EPIF-induced inhibition of Ca2+ pump. PMID- 9406157 TI - Cell-cycle dependent anti-FGF-2 staining of chicken cardiac myocytes: movement from chromosomal to cleavage furrow- and midbody-associated sites. AB - Fibroblast growth factor-2 (FGF-2) promotes cardiac myocyte proliferation and has been detected in extracellular as well as cytoplasmic and nuclear compartments. As a first step in examining the participation of intracellular FGF-2 in cardiac myocyte cell cycle we have investigated its localization in proliferative chicken cells during interphase and the various stages of mitosis in culture. We have used a previously characterized and affinity-purified anti-FGF-2 antibody preparation which recognizes the 19-22 kDa variants of chick FGF-2. By immunofluorescence, bright, punctate anti-FGF-2 labelling was observed in 26% of interphase nuclei from myocytes derived from 5 day embryonic heart ventricles; these nuclei were positive for anti-bromodeoxyuridine staining indicating that they are at the S- or G2 phase of the cell cycle. In prophase and metaphase, bright anti-FGF-2 staining was detected in apparent association with chromosomes. During anaphase, however, anti-FGF-2 staining dissociated from chromosomal locations distinctly remaining in strand-like structures in the area of ensuing cleavage furrow formation. In late telophase and cytokinesis, strong staining persisted in the area of the midbody and reappeared in a small fraction of newly formed daughter nuclei. Absorption of the antibody preparation with immobilized FGF-2 eliminated all staining. This dynamic pattern of anti-FGF-2 staining suggests that chick FGF-2 or immunologically related protein(s) not only increase in DNA-synthesizing nuclei but they may play a role in subsequent stages of mitosis and cytokinesis. PMID- 9406159 TI - Prostaglandins attenuate cardiac contractile dysfunction produced by free radical generation but not by hydrogen peroxide. AB - The aim of this study was to examine and compare the potential influence of cyclooxygenase or lipoxygenase derived metabolites of arachidonic acid on myocardial injury produced either by a free radical generating system consisting of purine plus xanthine oxidase or that produced by hydrogen peroxide. A free radical generating system consisting of purine (2.3 mM) and xanthine oxidase (10 U/L) as well as hydrogen peroxide (75 microM) produced significant functional changes in the absence of either significant deficits in high energy phosphates or ultrastructural damage. Prostaglandin F2 alpha (30 nM) significantly attenuated both the negative inotropic effect of purine plus xanthine oxidase as well as the ability of the free radical generator to elevate diastolic pressure. An identical concentration of prostaglandin 12 (prostacyclin) significantly reduced diastolic pressure elevation only and had no effect on contractile depression. The salutary effects of the two PGs occurred in the absence of any inhibitory influence on superoxide anion generation produced by the purine and xanthine oxidase reaction. None of prostaglandins modulated the response to hydrogen peroxide. In addition, neither prostaglandin E2 nor leukotrienes exerted any effect on changes produced by either type of oxidative stress. A 5 fold elevation in the concentrations of free radical generators or hydrogen peroxide produced extensive injury as characterized by a virtual total loss in contractility, 400% elevation in diastolic pressure, ultrastructural damage and significant depletions in high energy phosphate content. None of these effects were modulated by eicosanoid treatment. Our results therefore demonstrate a selective ability of both prostaglandin F2 alpha and to a lesser extent prostacyclin, to attenuate dysfunction produced by purine plus xanthine oxidase but not hydrogen peroxide. It is possible that these eicosanoids may represent endogenous protective factors under conditions of enhanced oxidative stress associated with superoxide anion generation. PMID- 9406160 TI - Changes in fatty acid compositions of myocardial lipids in rats with heart failure following myocardial infarction. AB - Changes in fatty acid composition of myocardial lipids were examined in rats with heart failure following myocardial infarction. Left ventricular systolic pressure (LVSP) was decreased and left ventricular end-diastolic pressure (LVEDP) was elevated 24 h, 1 and 12 weeks after left coronary artery ligation (CAL), suggesting the development of heart failure at these periods in this model. Hearts were isolated 24 h, 1 week and 12 weeks after the operation. Myocardial lipids in the infarcted scar tissue, non-infarcted remaining left ventricle including interseptum and right ventricle were separated into phospholipid (PL), triacylglycerol (TG), diacylglycerol (DAG) and free fatty acid (FFA) fractions. In the scar tissue PL content markedly decreased whereas TG, DAG and FFA contents increased 24 h after CAL. Despite a marked decrease in constituted fatty acids of PL fraction in the scar tissue the percentage of arachidonic acid in PL was elevated 12 weeks after CAL, suggesting that release of arachidonic acid during PL degradation was suppressed. In the non-infarcted viable left ventricle PL content remained unchanged throughout the experiment whereas TG, DAG and FFA contents were elevated 24 h after CAL. Despite no changes in PL and other lipid contents in the non-infarcted tissue the percentage of linoleic acid in PL was reduced and that of docosahexaenoic acid in PL was elevated 12 weeks after CAL. Our findings showed that myocardial lipid composition of the non-infarcted left ventricle was altered only in an early stage of the development of heart failure and fatty acid compositions of PL was exchanged in a late stage of the development of heart failure. The exchange may be related to cardiac dysfunction or myocardial remodelling in the rat with heart failure. PMID- 9406162 TI - Increases of T-type Ca2+ current in heart cells of the cardiomyopathic hamster. AB - In the present study, the whole-cell voltage clamp technique was used in order to record the T- and L-type Ca2+ currents in single heart cells of newborn and young normal and hereditary cardiomyopathic hamsters. Our results showed that the I/V relationship curve as well as the kinetics of the L-type Ca2+ currents (ICa(L)) in both normal and cardiomyopathic heart cells were the same. However, the proportion of myocytes from normal heart hamster that showed L-type ICa was less than that of heart cells from cardiomyopathic hamster. The I/V relationship curve of the T-type ICa (ICa(T)) was the same in myocytes of both normal and cardiomyopathic hamsters. The main differences between ICa(T) of cardiomyopathic and normal hamster are a larger window current and the proportion of ventricular myocytes that showed this type of current in cardiomyopathic hamster. The high density of ICa(T) as well as the large window current and proportion of myocytes showing ICa(T) may explain in part Ca2+ overload observed in cardiomyopathic heart cells of the hamster. PMID- 9406161 TI - Mechanisms that may be involved in calcium tolerance of the diabetic heart. AB - In diabetes the hearts exhibit impaired membrane functions, but also increased tolerance to Ca2+ (iCaT) However, neither the true meaning nor the molecular mechanisms of these changes are fully understood. The present study is devoted to elucidation of molecular alterations, particularly those induced by non-enzymatic glycation of proteins, that may be responsible for iCaT of the rat hearts in the stage of fully developed, but still compensated diabetic cardiomyopathy (DH). Insulin-dependent diabetes (DIA) was induced by a single i.v. dose of streptozotocin (45 mg.kg-1). Beginning with the subsequent day, animals obtained 6 U insulin daily. Glucose, triglycerides, cholesterol and glycohemoglobin were investigated in blood. ATPase activities, the kinetics of activation of (Na,K) ATPase by Na+ and K+, further the fluorescence anisotropy of diphenyl-hexatriene as well as the order parameters of membranes in isolated heart sarcolemma (SL) were also investigated. In addition, the degree of glycation and glycation related potency for radical generation in SL proteins were determined by investigating their fructosamine content. In order to study calcium tolerance of DH in a 'transparent' model, hearts were subjected to calcium paradox (Ca-Pa, 3 min of Ca2+ depletion; 10 min of Ca2+ repletion). In this model of Ca(2+) overload, Ca2+ ions enter the cardiac cells in a way that is not mediated by receptors. Results revealed that more than 83% of the isolated perfused DH recovered, while the non-DIA control hearts all failed after Ca-Pa. DH exhibited well preserved SL ATPase activities and kinetics of (Na,K)-ATPase activation by Na+, even after the Ca-Pa. This was considered as a reason for their iCaT. Pretreatment and administration of resorcylidene aminoguanidine (RAG 4 or 8 mg.kg 1) during the disease prevented partially the pathobiochemical effects of DIA induced glycation of SL proteins. DIA-induced perturbations in anisotropy and order parameters of SL were completely prevented by administration of RAG (4 mg.kg-1). Although, the latter treatment exerted little influence on the (Na,K) ATPase activity, it decreased the calcium tolerance of the DH. Results are supporting our hypothesis that the glycation-induced enhancement in free radical formation and protein crosslinking in SL may participate in adaptive mechanisms that may be also considered as 'positive' and are responsible for iCaT of the DH. PMID- 9406163 TI - Myocardial functional preservation during ischemia: influence of beta blocking agents. AB - To determine whether prior acute Beta blockade protects the heart against the deleterious effects of normothermic low flow global ischemia on myocardial function, aortic pressure, developed pressure, dP/dtmax and end diastolic pressure were monitored in isolated perfused rabbit hearts prior to, during and following 30 and 60 min ischemia, during which either Krebs-Henseleit (control) or Beta blocking agents. Bevantolol (cardioselective) or Propranolol (non selective) were perfused through the heart. Control hearts made ischemic for 30 min and then reperfused had significantly elevated end diastolic (p < .01) and aortic pressures (p < .01) and reduced developed pressure relative to baseline (p < .05). Hearts treated with Bevantolol or Propranolol (3 x 10(-5) m/l) 5 min prior to and during 30 min ischemia recovered preischemic developed pressure and dP/dtmax (p > 0.05), while end diastolic pressure was elevated (p < .01, p < .05 respectively). Aortic pressure was unchanged relative to baseline (p > .05). Comparison of indices from hearts under Beta blockade with controls showed that following 30 min ischemia and recovery, the Bevantolol treated group had reduced aortic pressure (p < .01) and end diastolic pressure (p < .05) and increased percent developed pressure and percent dP/dtmax (p < .001) relative to control. In the propranolol treated group, end diastolic pressure was reduced and percent developed pressure (p < .01) and percent dP/dtmax (p < .001) were increased relative to unblocked hearts. Following 60 min ischemia and 30 min reperfusion, reduction in all functional indices occurred, however dP/dtmax was unchanged from baseline in the Propranolol and Bevantolol treated groups. Comparison between groups showed that the Bevantolol treated group had significantly better dP/dtmax and developed pressure (p < .05), whereas the Propranolol group shows no significant difference from baseline (p > .05) (K-H). We conclude that following short periods of ischemia, Beta blockade protects the heart from deleterious function effects of ischemia but that the protective effect is diminished in Bevantolol relative to Propranolol treatments following prolonged ischemia. The data indicates that the beneficial effects of Beta blockade in reducing ischemic induced damage occurs early during conditions of ischemia such as would be present in the setting of acute myocardial infarction. PMID- 9406164 TI - Phosphorylation by protein kinase C and the responsiveness of Mg(2+)-ATPase to Ca2+ of myofibrils isolated from stunned and non-stunned porcine myocardium. AB - Previously we showed in an in situ porcine model that the thiadiazinone derivative [+]EMD 60263, a Ca2+ sensitizer without phosphodiesterase III inhibitory properties, increased contractility more profoundly in stunned than in non-stunned myocardium. This finding was consistent with the observed leftward shifts of the pCa2+/Mg(2+)-ATPase curves of isolated myofibrils induced by [+]EMD 60263. The aim of the present investigation was to study the possible involvement of protein kinase C in the mechanism of reduced Ca2+ responsiveness of myofilaments during stunning. No differences were observed in the maximal activity of the Ca(2+)-stimulated Mg(2+)-ATPase and in the pCa50 of myofibrils isolated from non-stunned and stunned myocardium. After phosphorylation with [gamma-32P]-ATP and excess of purified rat brain protein kinase C, the myofibrils were separated on sodiumdodecylsulphate-polyacrylamide gelectrophoresis and the 32P incorporation counted by the Molecular Imager. Ca2+/ phosphatidylserine/sn 1,2 diolein-dependent 32P incorporation catalyzed by excess of purified rat brain protein kinase C in C-protein, TnT and TnI subunits did not show any differences between myofibrils from non-stunned and stunned myocardium. However, protein kinase C-induced phosphorylation of myofibrils isolated from ventricular myocardium of sham-operated pigs resulted in a marked leftward shift of the pCa50 from 6.03 +/- 0.04 to 6.44 +/- 0.06 (p < 0.05), while porcine heart cyclic AMP dependent protein kinase-induced phosphorylation resulted in an expected small rightward shift to 5.97, although statistical significance was not reached. Protein kinase C-induced phosphorylation also stimulated (80%) the maximal myofibrillar Mg(2+)-ATPase activity. [+]EMD 60263 (3 microM) produced a leftward shift of the myofibrillar pCa2+/Mg(2+)-ATPase curve which was unaffected by prior protein kinase C-induced phosphorylation. In conclusion, the findings with isolated myofibrils from myocardium of anaesthetized open-chest pigs indicate that protein kinase C might be involved in the mechanism of reduced Ca2+ responsiveness of myofilaments in stunned myocardium. However, at this stage no differences could be found between the maximal activity of the Ca(2+)-stimulated Mg(2+)-ATPase, the pCa50 and the degree of phosphorylation of myofibrils isolated from stunned and non-stunned myocardium. PMID- 9406165 TI - Is there a link between impaired glucose metabolism and protein kinase C activity in the diabetic heart? AB - The activity of the beta isoform of protein kinase C (PKC beta) is reduced in the diabetic heart. Since this isozyme has been implicated in insulin action, we tested the hypothesis that PKC beta contributes to the development of impaired glucose metabolism by the noninsulin-dependent diabetic heart. Exposure of the diabetic heart to buffer containing the protein kinase C activator, phorbol myristate acetate, increased PKC beta activity in the membrane. Associated with the improvement in PKC beta activity was a biphasic change in glucose metabolism. The initial phase was characterized by a breakdown in glycogen stores, a stimulation in glucose oxidation and a decrease in endogenous fatty acid oxidation. This was followed by a second phase in which the uptake of glucose was modestly stimulated. Nonetheless, since the phorbol ester did not overcome the diabetes-linked defect in pyruvate dehydrogenase, the increase in glycolytic flux was not associated with a rise in glucose oxidation. Consequently, nearly 50% of the triose units were diverted into lactate and pyruvate production and the generation of ATP from glucose was restricted. Since insulin promotes not only glucose uptake, but also glycogen synthesis and glucose oxidation, the phorbol ester and insulin effects are very different. Thus, the data do not support a role for PKC beta in the development of glucose metabolic defects in the hearts of noninsulin-dependent diabetic rats. PMID- 9406166 TI - A new technique of coronary artery ligation: experimental myocardial infarction in rats in vivo with reduced mortality. AB - In vivo models of myocardial infarction following coronary artery ligation in the rat still suffer from high early mortality and a low rate of success of myocardial infarction. This study investigated the possibility of reducing early mortality and increasing the rate of myocardial infarction by modifications of surgical techniques. Eighteen rats were divided into two groups: normal control (3 rats) and ligation (15 rats). The major modifications of surgical techniques used in this study include: (1) no exteriorization of the heart, (2) ligation of the origins of the branches rather than the main trunk of the left coronary artery, (3) removal of air from the chest after closure, (4) supplying oxygen immediately after extubation. Following surgery, the rats recovered uneventfully and 11 rats were alive after 16 weeks. One rat, with a large myocardial infarction, died 2 h after surgery. Early mortality (during surgery and 1 week after surgery) was 6.7% with a success rate of myocardial infarction of 85%. The left ventricle in the ligation group showed significant dilation relative to normal and sham-operated control hearts (317% of control hearts, p < 0.001). However, myocardial mass did not increase. The average infarct size was 33%. These results demonstrate that a reduction in early mortality and an increased success rate of myocardial infarction can be achieved by modifications of surgical techniques. PMID- 9406168 TI - A calcium stimulated cysteine protease involved in isoproterenol induced cardiac hypertrophy. AB - The purpose of this study was to test the relationship between biochemical and functional changes accompanying beta-agonist induced cardiac hypertrophy and the activation of a calcium stimulated cysteine protease. Because the ultrastructural and ionic changes accompanying beta-agonist induced cardiac hypertrophy are reminiscent of the actions of the calcium activated neutral protease, calpain, it was hypothesized that lowering calpain activity (by the use of an exogenous inhibitor(s)) would reduce the extent of hypertrophy. Rats (275-300 g) were randomly assigned to either a control, beta-agonist (iso) or cysteine protease inhibitor (E64c) group. Isoproterenol administration (1 mg/kg) resulted in changes for ventricular weight to body weight ratio (increases 19%), ventricular [RNA] (increases 105.6%), rate of pressure development (increases 22% for +dP/dt) and maximum developed left ventricular pressure (increases 19%) (p < 0.05) after 3 days. Calpain-like activity (assessed by microplate method) increased by 45% (p < 0.05), while [cAMP] returned to control levels (following a transient rise at 1 day; 606.03 +/- 124.1 pmol/g/wet/wt to 937.9 +/- 225 (p < 0.05)). E64c (administered 1 h prior to iso) reduced the extent of hypertrophy, from 19 to 12%, and prevented the increases in; total [RNA], left ventricular function, the initial [cAMP] increase and calpain-like activity. It is concluded that a calcium stimulated cysteine protease(s), such as calpain, may be involved in the biochemical and functional changes associated with isoproterenol induced cardiac hypertrophy. PMID- 9406167 TI - Adriamycin depresses in vivo and in vitro phosphatidylethanolamine N-methylation in rat heart sarcolemma. AB - Adriamycin, an effective anticancer chemotherapeutic agent, causes an insidious and delayed cardiotoxicity. Different subcellular abnormalities including calcium transport changes in the sarcolemma (SL) as well as downregulation of the adrenergic system have been shown to be associated with the development of this cardiomyopathy. Since both of these activities are influenced by phospholipid methylation, effects of adriamycin on the three catalytic sites of SL phosphatidylethanolamine N-methyltransferase were examined. Rats were administered with a cumulative dose of adriamycin (15 mg/kg) over 2 weeks and examined after 3 weeks. Vehicle injected animals served as controls. Dyspnea, high mortality rate, ascites and decrease in aortic and left ventricular systolic pressure, as well as increase in left ventricular end diastolic pressure were seen in the adriamycin group. Myocardial cell damage typical of adriamycin cardiomyopathy, i.e. sarcotubular swelling, vacuolization and myofibrillar drop out, was also apparent. Total methyl group incorporation into SL phosphatidylethanolamine using radiolabeled S-adenosyl-L-methionine as the donor was significantly depressed in the 3 week group at catalytic sites II and III. Decreased production of methylated intermediates, phosphatidyl-N monomethylethanolamine and phosphatidyl-N,N-dimethylethanolamine as well as phosphatidylcholine (PC) was seen. Depression of phosphatidylethanolamine N methylation was also noticed when SL, isolated from untreated hearts, was exposed in vitro to different concentrations (10, 100 and 1000 microM) of adriamycin. Inhibition of phosphatidylethanolamine N-methylation appears to be mediated by adriamycin-induced increase in the oxidative stress and may contribute in the pathogenesis of subcellular changes associated with this cardiomyopathy. PMID- 9406169 TI - Early fetal like slow Na+ current in heart cells of cardiomyopathic hamster. AB - Using the whole-cell voltage-clamp technique, early embryonic tetrodotoxin (TTX) and Mn(2+)-insensitive slow Na+ current was detected in 10-22 week old fetal human heart cells as well as in 1 day old and young cardiomyopathic hamster myocytes. This slow Na+ current in both heart cell preparations has the same kinetics and pharmacology. This type of slow Na+ current was absent in heart cells of newborn and young normal hamsters and became less present in myocytes of 19 and 22 week old human heart myocytes. Our results demonstrate that the slow Na+ channel does exist in early fetal human life and this type of channel continues to be functional after birth in myocytes of the hereditary cardiomyopathic hamster. PMID- 9406170 TI - Na(+)-H+ exchange inhibition at reperfusion is cardioprotective during myocardial ischemia-reperfusion; 31P NMR studies. AB - To help resolve the controversy as to whether or not Na(+)-H+ exchange is functioning during reperfusion of the ischemic myocardium we assessed the effects of dimethylamiloride (DMA, an amiloride analogue possessing selectivity for inhibition of the Na(+)-H+ exchanger) on cardiac function and intracellular pH during ischemia-reperfusion. Studies were performed in the presence of bicarbonate (modified Krebs-Henseleit buffer) or in the nominal absence of bicarbonate (HEPES buffer) in order to determine if similar cardioprotection and effects on intracellular pH were observed in the presence and absence of bicarbonate dependent transport processes. Isovolumic rat hearts were perfused in the Langendorff mode at a constant pressure of 80 mm Hg and subjected to 28 min total global ischemia at 37 degrees C. Intracellular pH was determined from the pH dependent shift of the inorganic phosphate peak in 31P nuclear magnetic resonance spectra. DMA (20 microM) was infused for either 2.5 min before ischemia, for the initial 5 min of reperfusion, or at both time intervals. DMA had no effect on the intracellular pH during ischemia. Intracellular pH returned to pre-ischemic levels within 2.5 min of reperfusion in bicarbonate buffer. This normalization of pH was slower in HEPES perfusate. In both bicarbonate and HEPES perfused hearts all drug dosing regimens caused a significant increase in the recovery of mechanical function after reperfusion and slowed the recovery of intracellular pH during reperfusion. These results suggest that the Na(+)-H+ exchanger is activated during reperfusion of the ischemic myocardium, that this activation of the exchanger contributes to ischemia-reperfusion induced cardiac dysfunction and that administration of an inhibitor of Na(+)-H+ exchange at reperfusion significantly attenuates the deleterious effects of exchanger activation. PMID- 9406171 TI - Increased gene expression of plasminogen activators and inhibitors in left ventricular hypertrophy. AB - In the early stages of left ventricular hypertrophy (LVH) acute adaptive changes occur in the coronary vasculature as it remodels. Plasminogen activators (PAs) and inhibitors (PAIs) have the potential effects of proteolytic degradation that is relevant to tissue remodeling and angiogenesis. Our study focused on the possible roles of PAI-1, PAI-2, and uPA in tPA in myocyte hypertrophy and angiogenesis in the early and late stages of pressure overload induced left ventricular hypertrophy (LVH). We divided seventeen adult swine, weighing 24.2 +/ 6.5 kg, into four groups: control, sham-operated, early LVH and late heart failure LVH group. At surgery we placed a fixed constrictor on the ascending aorta immediately above the aortic valve. This increased LV systolic pressure from 133 +/- 15 to 193 +/- 24 mm Hg after the surgery. We subdivided the early group into groups of 3 animals each that we euthanized at 8, 24 and 72 h after operation and obtained heart samples for analysis. In the late heart failure group individual animals were euthanized at 55, 59, 62 and 72 days after the detection of congestive heart failure. We also obtained tissue samples from the control and sham-operated swine. Sections for histologic analysis were fixed in 10% buffered formalin. We isolated RNA, size fractionated it using 1% formaldehyde-agarose gel electrophoresis and then did Northern blots. The mRNAs from both PAI-1 and PAI-2 showed a remarkable increase at 8 and 24 h after acute aortic constriction and returned to control by 72 h. Regional differences showed that most of the increases were in the endocardium. Three animals in the late heart failure LVH group were determined to be in congestive heart failure at about 2 months after the onset of aortic constriction. In these animals PAI-1 and PAI-2 were increased in both the left and right ventricles but remained low in an animal of the same elevation in aortic pressure seen by the LV who did not have congestive failure. These data suggest that PA and PAI gene expressions change before morphologic changes occur in the early stages of developing LVH. Also at the time of onset of congestive heart failure this increased expression reappears. PAs and PA inhibitors mRNA levels vary in the different regions of the heart reflecting changing wall stresses. Thus, the PAs and PA inhibitors may play an important role in angiogenesis that occurs during the early stages of LVH. The increased expression in the late stage of LVH may reflect further changes in wall stresses since these animals also showed overt clinical signs of heart failure. PMID- 9406172 TI - Cardiac hypertrophy: old concepts, new perspectives. AB - Growth of the heart in hypertrophy is accompanied by changes in the phenotypic expression of cardiac genes. To explore the molecular basis of cardiac hypertrophy, we have analyzed the regulation of myosin heavy chain gene (MHC) expression. In one set of experiments, pressure overload on the rat heart was produced by constriction of the abdominal aorta. Changes in the alpha and beta MHC mRNA were then studied in overloaded hearts and following load removal. Pressure overload resulted in down-regulation of the alpha-MHC with corresponding up-regulation of the steady state level of beta-MHC mRNA. Load removal (debanding) resulted in regression of cardiac hypertrophy and a rapid return of alpha-MHC mRNA to normal values. In contrast, the recovery in beta-MHC mRNA was much slower to the extent that it remained substantially elevated compared to respective sham controls even after 7 weeks of post-debanding. These results suggest that putative load-related signals independently regulate two genes. Several lines of evidence indicate that adrenergic nervous system plays an important role in the induction and maintenance of cardiac hypertrophy and in the redistribution of myosin isoforms. We have analyzed the effect of cAMP inducing agents on the regulation of alpha-MHC gene in primary cultures of the fetal (18 day) rat cardiac myocyte. Inclusion of 8 Br-cAMP in the culture media increased the expression of alpha-MHC promoter/reporter construct comprising of 2.9 kb upstream sequence of the alpha-MHC gene. Several deletion mutations in the alpha MHC gene promoter defined the cAMP responsive boundaries to be a 32 bp region comprising of -71 to -40 bp sequences. Deletion of this region resulted in loss of cAMP response as well as in basal expression of alpha-MHC promoter/reporter construct. These data suggest a role of beta-adrenergic pathway in the modulation of alpha-MHC gene expression. PMID- 9406173 TI - Characteristics of the myocardial PM-FABP: effect of diabetes mellitus. AB - Properties of the myocardial PM-FABP were studied in normal and STZ-diabetic rats. The fluorescent fatty acids trans-parinaric and cis-parinaric acids were used as analogs of straight-chain (saturated) and kinked-chain (unsaturated) fatty acids respectively. Parinaric acid binding was sensitive to trypsin. Trans parinaric acid binding was more sensitive to this protease than the binding of cis-parinaric acid. Based on the difference in sensitivity of parinaric acid binding we believe that there are two separate binding sites associated with myocardial PM-FABP; one for unsaturated fats and the other for saturated fats. Diabetes enhanced both cis- and trans-parinaric acid binding capacity in cardiomyocytes; cis-parinaric acid by 2 fold and trans-parinaric acid by 2.6 fold. In addition, there was a concomitant accumulation of free fatty acids and triglycerides in the hearts of the diabetic animals. There was a 2.2 fold increase for fatty acids and a 1.6 fold increase for trigylcerides. This association between myocardial fatty acid build-up and enhanced myocardial PM FABP during diabetes suggest that this carrier protein might have contributed to lipid accumulation in the hearts of the diabetic rats. PMID- 9406174 TI - Cardiomyopathies and mitochondrial DNA mutations. AB - Our former studies concerning mitochondrial DNA mutations were reviewed in this article. A 7.4 kb deletion between the D-loop and ATPase 6 genes was detected in myocardial tissue obtained at autopsy from patients with myocardial infarction, diabetes mellitus and also patients treated with adriamycin. A case with diabetes mellitus and hypertrophic cardiomyopathy is demonstrated which revealed a point mutation from adenine to guanine at position 3243 within tRNA Leu(UUR). PMID- 9406176 TI - Ischemic preconditioning is not additive to preservation with hypothermia or crystalloid cardioplegia in the globally ischemic rat heart. AB - The aim of this study was to evaluate the additive protective efficiency of ischemic preconditioning when used in combination with conventional clinically relevant cardioprotective methods of hypothermia or hypothermic cardioplegia during sustained global ischemia. Isolated rat hearts were aorta-perfused with Krebs-Henseleit buffer and were divided into six groups (n = 10 each). Group I: Ischemia at 34 degrees C for 60 min; Group PC + I: preconditioned (PC) ischemia at 34 degrees C, 2 episodes of 5 min ischemia and 10 min reperfusion at 34 degrees C followed by I; Group HI: hypothermic ischemia at 10 degrees C for 60 min; Group PC + HI: preconditioned (PC) hypothermic ischemia, 2 episodes of 5 min ischemia and 10 min reperfusion at 34 degrees C followed by HI; Group CPL + HI: single dose of 'Plegisol' cardioplegia followed by HI; Group PC + CPL + HI: preconditioned hypothermic cardioplegia, followed by CPL + HI. At the end of 60 min ischemia, all the hearts were reperfused at 34 degrees C for 30 min when post ischemic recovery in left ventricular contractile function and coronary vascular dynamics was computed and compared. There was a significant depression in the post-ischemic recovery of developed pressure (Pmax), positive derivative of pressure (+dp/dt), negative derivative of pressure (-dp/dt) and heterometric autoregulation (HA) of contractile force in all the groups, with no major differences between the groups. Left ventricular end-diastolic pressure (LVEDP) was significantly elevated after I at 34 degrees C. Preconditioning (PC + I) prevented the rise in the LVEDP and this was accompanied by a significant reduction in the release of purine metabolites in the coronary effluents, particularly adenosine, during the immediate reperfusion period. Hypothermia (HI) provided essentially the same level of metabolic and mechanical preservation as offered by PC + I. Combination of hypothermia with preconditioning (PC + HI) or cardioplegia (PC + CPL + HI), did not further enhance the preservation. Post ischemic recovery in the regional contractile function (segment shortening, %SS) followed nearly identical pattern to global (Pmax) recovery. Post-ischemic recovery in coronary flow (CF) was significantly reduced and coronary vascular resistance (CVR) was significantly increased in all the groups. Myogenic autoregulation (transient and sustained) was generally enhanced indicating increased vascular reactivity. Preconditioning did not alter the time-course of these changes. Preconditioned ischemia (34 degrees C) preserved left ventricular diastolic functions and prevented the contracture development after sustained ischemia reperfusion at 34 degrees C. This protective effect of preconditioning was possibly mediated by the reduction in the breakdown of purine metabolites. Hypothermia alone or in combination with crystalloid cardioplegia prevented the irreversibility of the ischemic injury but produced contractile and vascular stunning which was not improved by ischemic preconditioning. The results of this study indicate that preconditioning when combined with hypothermia or hypothermic cardioplegia offered no significant additional protection. PMID- 9406175 TI - Cardiac depression and cellular injury in hemorrhagic shock and reinfusion: role of free radicals. AB - We investigated the effect of hemorrhagic shock and reinfusion on the cardiac function and contractility, plasma CK and CK-MB activity and lactate concentration, oxyradical-producing activity of polymorphonuclear leukocytes (PMNL-CL), cardiac chemiluminescence (LV-CL), antioxidant enzyme activity [superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-PX)] and malondialdehyde (MDA) concentration in anesthetized dogs to determine the role of oxyradicals in cardiac depression and cellular injury in hemorrhagic shock and reinfusion. The dogs were assigned into three groups: I (sham), 4 h duration; II (S + R), 2 h of shock followed by reinfusion for 2 h; III (SOD + S + R), as II but pretreated with PEG-SOD. Hemorrhagic shock was produced by withdrawal of blood to maintain the mean arterial pressure at 50 +/- 5 mm Hg. Cardiac function and contractility were depressed during hemorrhagic shock. Plasma CK, CK-MB and lactate increased during shock. Following reinfusion after 2 h of shock hemodynamic parameters and plasma lactate tended to return towards control values. Plasma CK and CK-MB, PMNL-CL and cardiac MDA, total-, Mn- and CuZn-SOD activity increased while LV-CL decreased. In spite of the increase in the antioxidant reserve, there was oxidative damage. Pretreatment with SOD attenuated the deleterious effects of shock and reinfusion on the cardiovascular function, plasma CK, and CK-MB, PMNL-CL, cardiac MDA, SOD, and LV-CL. Protection was incomplete for cardiovascular function and plasma CK and CK-MB. These results suggest that oxyradicals may partly be involved in the deterioration of cardiovascular function and cellular injury during hemorrhagic shock and reinfusion. PMID- 9406178 TI - Age- and sex-related differences in nuclear lipid content and nucleoside triphosphatase activity in the JCR:LA-cp corpulent rat. AB - The putative role of the nuclear nucleoside triphosphatase (NTPase) is to provide energy to the nuclear pore complex for poly A(+) mRNA export. Previous work has demonstrated that liver nuclear NTPase activity is greater in 6 month old corpulent (cp/cp) female JCR:LA rats, a hyperlipidemic rat model, compared to lean (+/?) animals. This increase appeared to be related to increases in nuclear membrane cholesterol content. The current study extended these initial data to compare NTPase activity as a function of age and sex in isolated JCR:LA-cp rat liver nuclei, to further test the hypothesis that nuclear membrane cholesterol may modulate NTPase activity. NTPase activity was increased in cp/cp female animals compared to +/? females at all ages studied, with Vmax values increased by 60-176%. Membrane integrity of cp/cp female nuclei was reduced compared to +/? female nuclei. Nuclear membrane cholesterol levels increased linearly with age by 50, 150 and 250% in 3, 6 and 9 month old cp/cp females over leans. In contrast, nuclei from cp/cp males exhibited only minor, isolated changes in NTPase activity. Furthermore, there were no significant changes in nuclear cholesterol content or membrane integrity in the less hyperlipidemic male animals at any age. These data suggest that altered lipid metabolism may lead to changes in nuclear membrane structure, which in turn may alter NTPase activity and functioning of the nuclear pore complex. PMID- 9406177 TI - Regulation of Ca2+ homeostasis by glucose metabolism in rat brain. AB - In a previous communication we reported that glucose deprivation from KHRB medium resulted in a marked stimulation of Ca2+ uptake by brain tissue, suggesting a relationship between glucose and Ca2+ homeostasis in brain tissue. Experiments were carried out to investigate the significance of glucose in Ca2+ transport in brain cells. The replacement of glucose with either D-methylglucoside or 2 deoxyglucose, non-metabolizable analogues of glucose, resulted in stimulation of Ca2+ uptake just as by glucose deprivation. These data show that glucose metabolism rather than glucose transfer was necessary to stimulate Ca2+ uptake in brain tissue. Inhibition of glucose metabolism with either NaF, NaCN, or iodoacetate resulted in stimulation of Ca2+ uptake similar to that produced by glucose deprivation. These results lend further support for the concept that glucose metabolism is essential for Ca2+ homeostasis in brain. Anoxia promotes glucose metabolism through glycolytic pathway to keep up with the demand for ATP by cellular processes (the Pasteur effect). Incubation of brain slices under nitrogen gas did not alter Ca2+ uptake by brain tissue, as did glucose deprivation and the inhibitors of glucose metabolism. We conclude that glucose metabolism resulting in the synthesis of ATP is essential for Ca2+ homeostasis in brain. Verapamil and nifedipine which block voltage-gated Ca2+ channels, did not alter Ca2+ uptake stimulated by glucose deprivation, indicating that glucose deprivation-enhanced Ca2+ uptake was not mediated by Ca2+ channels. Tetrodotoxin which specifically blocks Na2+ channels, abolished Ca2+ uptake enhanced by glucose deprivation, but had no effect on Ca2+ uptake in presence of glucose (controls). These results suggest that stimulation of Ca2+ uptake by glucose deprivation may be related to Na2+ transfer via NaCa exchange in brain. PMID- 9406180 TI - An overview of clinical molecular genetics. AB - Clinical molecular genetics has recently become recognized as a diagnostic discipline. This article covers the evolution, structure, and possible forward development of clinical molecular genetics. Topics covered include general test categories, introducing new tests, laboratory facilities, staffing and training, and overview of quality issues. PMID- 9406179 TI - Studies on hepatic injury and antioxidant enzyme activities in rat subcellular organelles following in vivo ischemia and reperfusion. AB - The activities of rat hepatic subcellular antioxidant enzymes were studied during hepatic ischemia/reperfusion. Ischemia was induced for 30 min (reversible ischemia) or 60 min (irreversible ischemia). Ischemia was followed by 2 or 24 h of reperfusion. Hepatocyte peroxisomal catalase enzyme activity decreased during 60 min of ischemia and declined further during reperfusion. Peroxisomes of normal density (d = 1.225 gram/ml) were observed in control tissues. However, 60 min of ischemia also produced a second peak of catalase specific activity in subcellular fractions corresponding to newly formed low density immature peroxisomes (d = 1.12 gram/ml). The second peak was also detectable after 30 min of ischemia followed by reperfusion for 2 or 24 h. Mitochondrial and microsomal fractions responded differently. MnSOD activity in mitochondria and microsomal fractions increased significantly (p < 0.05) after 30 min of ischemia, but decreased below control values following 60 min of ischemia and remained lower during reperfusion at 2 and 24 h in both organelle fractions. Conversely, mitochondrial and microsomal glutathione peroxidase (GPx) activity increased significantly (p < 0.001) after 60 min of ischemia and was sustained during 24 h of reperfusion. In the cytosolic fraction, a significant increase in CuZnSOD activity was noted following reperfusion in animals subjected to 30 min of ischemia, but 60 min of ischemia and 24 h of reperfusion resulted in decreased CuZnSOD activity. These studies suggest that the antioxidant enzymes of various subcellular compartments respond to ischemia/reperfusion in an organelle or compartment specific manner and that the regulation of antioxidant enzyme activity in peroxisomes may differ from that in mitochondria and microsomes. The compartmentalized changes in hepatic antioxidant enzyme activity may be crucial determinant of cell survival and function during ischemia/reperfusion. Finally, a progressive decline in the level of hepatic reduced glutathione (GSH) and concomitant increase in serum glutamate pyruvate transaminase (SGPT) activity also suggest that greater tissue damage and impairment of intracellular antioxidant activity occur with longer ischemia periods, and during reperfusion. PMID- 9406182 TI - Overview of vaccines. AB - This article lists the vaccines current available for the control of both viral and bacterial infections. They may be attenuated live or inactivated whole microorganisms, or subunit preparations. Many more are in the pipeline and increasing attention is being given to establishing their safety before registration. Following the earlier eradication of smallpox, good progress is now being made toward the global eradication of poliomyelitis and a new program to eliminate measles from the Americas has begun. A variety of new approaches to vaccine development is now available. The hepatitis B virus surface antigen, made by DNA-transfected yeast or mammalian cells, is the basis of the first genetically engineered vaccine. Early in the 21st century, new vaccines based on oligopeptides, recombinant live viral or bacterial vectors (often existing live vaccines), or recombinant DNA plasmids are likely to be registered for human use. The efficacy of vaccines depends on the immune responses generated, and the recent substantial increase in our understanding of the mammalian immune system now offers great opportunities for manipulation to best obtain desired responses. These include mixing vaccine formulations to maximize immune responses, and combining vaccines to simplify their administration. Despite these advances, some persisting infections, such as those caused by HIV, plasmodia, and mycobacteria, still pose a great challenge to vaccine developers. PMID- 9406181 TI - Degradative covalent reactions important to protein stability. AB - Commonly observed chemical modifications that occur in proteins during their in vitro purification, storage, and handling are discussed. Covalent modifications described include deamidation and isoaspartate formation, cleavage of peptide bonds at aspartic acid residues, cystine destruction and thiol-disulfide interchange, oxidation of cysteine and methionine residues, and the glycation and carbamylation of amino groups. PMID- 9406183 TI - An effective method of completely removing contaminating genomic DNA from an RNA sample to be used for PCR. AB - A simple method for removing contaminating genomic DNA from an RNA preparation is presented. The method involves digestion of the RNA with RNase-free DNase I at room temperature followed by inactivation of the enzyme at 65 degrees C in presence of EDTA. This method produces an RNA sample that is negative for genomic DNA by PCR. PMID- 9406184 TI - Identification of microorganisms using random primed PCR. AB - The polymerase chain reaction has facilitated the use of molecular approaches in microbiology including new strategies for the rapid identification of micro organisms. Approaches based on the use of random primers and standard conditions, allows characteristic DNA fingerprints to be generated from any micro-organism even in the absence of information about its DNA sequence. Different primers can be used to produce genus-specific, species-specific, or even strain-specific DNA fingerprints. This article covers the background to this strategy, describes three different approaches to generating DNA fingerprints using random primers, and provides experimental detail for one method, RAPD. PMID- 9406185 TI - Generation of large insert yeast artificial chromosome libraries. AB - The development of YAC cloning technology has directly enhanced the relationship among genetic, physical, and functional mapping of genomes. Because of their large size, YACs have enabled the rapid construction of physical maps by ordered clone mapping and contig building, and they complement other molecular approaches for mapping complex genomes. Large insert libraries are constructed by size fractionating large DNA embedded in agarose and protecting DNA from degradation with polyamines. PMID- 9406187 TI - Measurement of immunoglobulin synthesis using the ELISPOT assay. AB - We describe a method for the detection of specific antibody-producing cells from either in vitro or in vivo immunization. These techniques are especially useful for detecting antibodies from developing hybridomas. We have successfully used the system to detect isotype-specific antibodies to a variety of bacterial antigens which were produced by heterohybridomas. PMID- 9406186 TI - Enzymatic cleavage and HPLC peptide mapping of proteins. AB - Detailed procedures are described for successfully digesting reasonably small quantities (i.e., usually > 10 pmol) of proteins with a variety of proteases and for then isolating the resulting peptides by reverse-phase HPLC. Since sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) appears to be the current method of choice for final purification of proteins for structural analysis, special attention is given to carrying out in-gel proteolytic digests on SDS-PAGE-separated proteins that have usually been stained with Coomassie Blue. A compilation of data from nearly 200 "unknown" samples is used to help provide realistic expectations with respect to the results that are likely to be obtained from carrying out in-gel proteolytic digests on large numbers of proteins. PMID- 9406188 TI - Expression of human immunodeficiency virus genes using baculovirus expression system. AB - The structural protein genes of HIV-1 and HIV-2 have been expressed in Spodoptera frugiperda (SF) cells using baculovirus expression system. The noncoding flanking sequences of HIV structural genes were removed and a putative ribosome binding site was placed in front of the open reading frame of each gene by using crossover linker mutagenesis. The coding sequences of the gag, pol, env, and vif proteins were inserted into Autographa californica nuclear polyhedrosis virus (AcNPV) so that HIV genes were under the control of the AcNPV polyhedrin promoter. All recombinant AcNPV-infected SF cells express high levels of HIV structural proteins. Detailed strategies of recombinant AcNPV construction for high level protein expression are presented. PMID- 9406189 TI - The sacB gene cannot be used as a counter-selectable marker in Pasteurella multocida. AB - The use of the Bacillus subtilis sacB gene as a counter-selectable marker was assessed in serogroup A and B strains of Pasteurella multocida. Expression ofsacB failed to render any of the strains sensitive to sucrose, indicating that the sacB gene can not be used as a positive selection system in P. multocida. PMID- 9406191 TI - Androgenesis and homozygous gynogenesis in muskellunge (Esox masquinongy): evaluation using flow cytometry. AB - The purpose of this work was to study the effects of ultraviolet (UV) irradiation on denucleation of eggs and investigate the heat-shock conditions for diploidization for induction of androgenesis in muskellunge, Esox masquinongy. Several egg incubation media, including saline, Ringer's solution, and Ringer's solution supplemented with bovine serum albumin (BSA), were found suitable to maintain the egg fertility as high as in muskellunge ovarian fluid. The optimal doses of UV radiation were 660-1320 J/m2, at which 100% haploid larvae were produced at a hatching rate of 22.5 +/- 2.8%. UV irradiation at low doses (165 330 J/m2) generated abnormal larvae, which were morphologically identical to haploids. Using a flow cytometry method, it was found that cellular DNA content of these larvae was close to that of diploids but significantly lower in value and had a wider distribution (expressed as coefficient of variation) than that of control fish. This suggested that a low dose of UV irradiation might cause gene mutations, alteration of chromosomal conformation and fragmentation, but did not prevent maternal DNA from participating in mitotic division. Interference of maternal DNA residues could be another reason for the poor viability of androgenetic fish. A high dose of UV radiation (1980 J/m2) caused development of severely deformed embryos, indicating that UV radiation also damaged molecules in the eggs other than the denucleation. Our results suggest that classic color and allozyme markers might not be sufficient to prove a complete androgenesis. In order to optimize time and duration of shock for induced diploidization, we investigated the heat-shock conditions for inhibiting the first mitotic cleavage through induction of homozygous gynogenesis. We found that heat-shock treatment at 31 degrees C for 9 min starting at 1.4 tau 0 (a dimensionless factor describing progress in embryo development) after fertilization produced the highest percentage of diploids at hatching. PMID- 9406190 TI - Molecular cloning, genetic mapping, and developmental expression of a bovine transforming growth factor beta (TGF-beta) type I receptor. AB - A full-length cDNA encoding the bovine transforming growth factor beta (TGF-beta) receptor type I (bT beta R-I) was isolated from a placenta cDNA library. The deduced protein sequence of 499 residues contains a single transmembrane domain, a cysteinerich extracellular domain, and an intracellular kinase domain with predicted serine/threonine specificity. The amino acid sequence is 96% and 95% identical with its human and mouse homologues, respectively. Genetic mapping assigned the TGFBR1 gene to bovine chromosome 8 at a male genetic distance of 2 centimorgan from D8S28. Assuming conservation of gene order, the linkage data define a breakpoint in mammalian chromosome evolution. Both TGF-beta receptor type I and II mRNAs were found to be expressed in bovine oocytes and preimplantation two-cell, four-cell, eight-cell, morula-, and blastocyst-stage embryos, as determined by heminested reverse transcription polymerase chain reaction (RT-PCR). The mRNA expression patterns of TGF-beta receptor types I, II, and III in a variety of bovine organ tissues were examined by Northern blot hybridization, and highest levels were detected in lung and ovary. PMID- 9406192 TI - The stage-specific expression of TEC-1, -2, -3, and -4 antigens on bovine preimplantation embryos. AB - The preimplantation developmental period is associated with constant changes within the embryo, and some of these changes are apparent on the embryo cell surface. For example, during transition from maternal to embryonic genome control and the compaction and differentiation of embryonic cells, the cell surface undergoes morphologic alterations that reflect changes in gene control. In order to gain insight into the events occurring during embryonic development and cellular differentiation, monoclonal antibodies specific for cell surface antigens (TEC antigens) of embryonic cells have been generated previously and shown to recognise either the carbohydrate moiety of embryoglycan or a developmentally regulated protein epitope. The TEC antigens have been identified on mouse preimplantation embryos, and their expression is specific to particular developmental stages. To determine whether these antigens are conserved in higher mammals, we examined the expression of four TEC antigens (TEC-1 to TEC-4) on in vitro-derived bovine and murine embryos during the preimplantation stage of development. It was found that bovine oocytes and embryos derived from in vitro maturation (IVM) and in vitro fertilisation (IVF) showed stage-specific expression of each of the TEC antigens investigated, with the pattern of expression overlapping but not identical to that seen in the mouse. Immunoprecipitation together with Western blot analysis showed that the TEC monoclonal antibodies recognised a single glycoprotein band with an apparent molecular weight of 70 kDa. Confocal microscopy of immunofluorescence staining of the bovine cells showed this protein to be located on the cell surface. The apparent negative expression of these TEC antigens by immunohistochemistry and immunoprecipitation at particular stages of development appears to be due to the epitopes being inaccessible to the TEC antibodies, since Western blotting revealed the TEC antigens to be present at all stages of development examined. Antibodies identifying stage-specific antigens will provide useful markers to characterise early embryonic cells, monitor normal embryonic development in vitro, and identify cell surface structures having a function in cell-cell interactions during embryogenesis and differentiation. PMID- 9406193 TI - Telophase enucleation: an improved method to prepare recipient cytoplasts for use in bovine nuclear transfer. AB - The enucleation of oocytes to be used as host cytoplasts for embryo reconstruction by nuclear transfer is an important limiting step when cloning mammals. We propose an enucleation technique based on the removal of chromatin after oocyte activation, at the telophase stage, by aspirating the second polar body and surrounding cytoplasm. In a preliminary experiment to determine an optimal activation protocol, oocytes were matured for 26 and 30 hr and exposed for 5 min to 7% ethanol and/or for 3 hr at either 25 or 4 degrees C. Relative to most activation treatments tested, oocytes matured for 30 hr and exposed to ethanol alone showed highest activation rates, as determined by low levels of H1 kinase activity within 90 min from exposure and high pronuclear formation (82%) after 12 hr of culture. No synergistic effect on activation rates was observed when oocytes also were exposed to reduced temperature after ethanol treatment. Microsurgical removal of the telophase-stage chromatin in a small volume of cytoplasm adjacent to the second polar body was significantly more effective in enucleating than aspiration of a larger cytoplasm volume surrounding the first polar body of metaphase-arrested oocytes (98% versus 59%; P < 0.01). Moreover, compared with a nuclear transfer protocol based on enucleation of metaphase arrested oocytes followed by aging and cooling, more (38% versus 16%; P < 0.001) and better-quality blastocytes (126 versus 84 nuclei per blastocyst; P < 0.02) were obtained from embryos reconstructed using the telophase procedure. Higher development potential of embryos reconstructed by the telophase procedure may be attributed to (1) the selection of oocytes that activate and respond by extruding the second polar body, (2) avoiding the use of DNA dyes and ultraviolet irradiation, and (3) the limited removal of cytoplasm during enucleation. The ease with which telophase enucleation can be performed is likely to render this technique widely useful for research and practice on mammalian cloning. PMID- 9406194 TI - Injection of a porcine sperm factor induces activation of mouse eggs. AB - Injection of sperm preparations into mammalian oocytes and eggs has been shown to elicit persistent [Ca2+]i oscillations that closely resemble fertilization associated Ca2+ release. However, the ability of these sperm fractions to initiate egg activation has not been clearly demonstrated. In the present experiments, mouse eggs injected with a porcine sperm preparation were evaluated for early and late events of activation. Events monitored included, among early events, the generation of [Ca2+]i oscillations and cortical granule exocytosis and, among late events, the decrease in histone H1 and myelin basic protein kinase activities, polar body extrusion, pronuclear formation, and cleavage to the two-cell stage. Injection of sperm fractions consistently evoked [Ca2+]i oscillations that, in turn, initiated all events of activation. Uninjected control eggs or eggs injected with buffer or heat-treated sperm fractions failed to show Ca2+ responses or activation. In addition, injection of sperm fractions into recently ovulated eggs (experiments were concluded within 15 hr after human chorionic gonadotropin administration) induced high rates of activation, while similarly aged eggs exposed to 7% ethanol for 5 min, a known parthenogenetic treatment, failed to activate. Together these results indicate that injection of sperm fractions elicits [Ca2+]i oscillations that are capable of initiating normal egg activation. These results support the hypothesis that a sperm component participates in the generation of fertilization-associated [Ca2+]i oscillations. PMID- 9406195 TI - Inhibition of domestic cat spermatozoa acrosome reaction and zona pellucida penetration by tyrosine kinase inhibitors. AB - Spermatozoa from teratospermic domestic cats (> 60% morphologically abnormal spermatozoa per ejaculate) consistently exhibit lower levels of oocyte penetration in vitro than their normospermic (< 40% abnormal spermatozoa per ejaculate) counterparts. This could be caused by structural abnormalities or intracellular defects resulting in disruption of normal cellular functions. Spermatozoa from teratospermic cats also are compromised in the ability to capacitate and undergo the acrosome reaction (AR) in vitro. Further, we recently identified two tyrosine phosphorylated proteins (95- and 160-kDa) localized over the acrosome region in domestic cat spermatozoa. Phosphorylation of these proteins is reduced in teratospermic compared with normospermic ejaculates. To begin to understand the relationship between tyrosine phosphorylation and sperm function, we examined the effects of two protein tyrosine kinase inhibitors (tyrphostin RG-50864 and genistein) on (1) sperm motility; (2) protein tyrosine phosphorylation; (3) the ionophore A23187-induced AR; (4) the spontaneous and zona pellucida (ZP)-induced AR, and (5) the ability of spermatozoa from normospermic cats to penetrate conspecific ZP-intact oocytes. Over a wide range of concentrations, neither inhibitor affected sperm percentage motility during incubation (P > 0.05). Preincubation with either inhibitor reduced tyrosine phosphorylation of both (95- and 160-kDa) sperm proteins. Although both inhibitors blocked the ZP-induced AR, neither influenced the spontaneous AR nor the A23187-induced AR, suggesting that tyrosine phosphorylation may be involved in physiologic AR. No differences (P > 0.05) were observed in the ability of control or inhibitor-treated spermatozoa to bind to or penetrate the outer ZP layer. However, percentages of oocytes with treated spermatozoa in the inner ZP (tyrphostin, 8.7%; genistein, 20.4%) and perivitelline space (tyrphostin, 0%; genistein, 2.3%) were less (P < 0.001) than untreated controls (inner ZP, 62.7%; perivitelline space, 10.2%). These results (1) demonstrate that ZP-induced acrosomal exocytosis in domestic cat spermatozoa is regulated via a tyrosine kinase-dependent pathway and (2) suggest that defects in these signaling pathways may represent one of the causes for compromised sperm function in teratospermic males. PMID- 9406196 TI - Identification and spatial distribution of the mRNA encoding the gp49 component of the gilthead sea bream, Sparus aurata, egg envelope. AB - A cDNA encoding the precursor of one of the major components of gilthead sea bream, Sparus aurata, egg envelope has been cloned by reverse transcriptase polymerase chain reaction (RT-PCR) techniques. The clone was isolated starting from total RNA extracted from the liver of spawning female fish and estradiol-17 beta-treated male fish. Sequence analysis revealed that the cDNA encoded a protein of 405 aa corresponding to 49-kDa component (termed gp49), a glycoprotein belonging to the N-linked type. The gp49 protein is homologous to the Zl-3 of medaka Oryzias latipes, the mammalian ZPC and ZPC homologues of Xenopus laevis (xlZPC) and carp Cyprinus carpio (ccZPC). In addition, the open reading frame also encodes an additional aa sequence, the signal peptide, located in the N terminal region of the protein. RT-PCR and in situ expression analyses evidenced an organ-restricted pattern: the mRNA was detected only in liver of spawning female and estradiol-17 beta-treated male fish but not in other tissues. PMID- 9406197 TI - Chromatin configuration during meiosis I prophase of spermatogenesis. AB - During the pachytene stage of meiotic prophase in male mammals, the X and Y chromosomes become transcriptionally inactive and establish a chromatin domain, the sex body, that is visually distinct from the transcriptionally active autosomes. We used objective criteria to assess these chromatin differences by DNase I sensitivity (DS) of sex chromosome and autosomal sequences at both the cytological and molecular levels. For cytological studies, in situ nick translation techniques were used on air-dried preparations of testicular cells. For molecular studies, nuclei from pachytene spermatocytes were subjected to nuclease sensitivity assays. Both sex-linked and autosomal sequences were assessed, including some gene sequences that are expressed and some that are not expressed in pachytene spermatocytes. There was a wide range of DS in different genomic sequences; however, the sex-linked sequences generally were less nuclease sensitive than were autosomal sequences. Interestingly, a hot spot of recombination (within the Eb gene) showed a high level of nuclease sensitivity, while a cold spot of recombination (centromeric satellite region) exhibited lower sensitivity, more similar to that of sex-linked sequences. We also examined the nuclease sensitivity of a tyrosinase transgene insert, TyBS. In one line of mice, the transgene insert is X-linked, whereas in another, it is autosomal. The transgene was less nuclease sensitive when X-linked than as an autosomal insert. These results support the hypothesis that in pachytene spermatocytes the XY chromosome pair is more condensed and inaccessible to enzymatic digest, whereas the autosomal chromatin is in a more open configuration. In addition, we examined the nuclease sensitivity of some of the same genes in the earlier leptotene/zygotene prophase stage, when the sex chromatin is not maximally condensed. We found that while autosomal gene nuclease sensitivity was equivalent to that at the pachytene stage, X-linked sequences were more nuclease sensitive. Overall, these differences in chromatin nuclease sensitivity correlate with differences in meiotic recombination activity and may be mechanistically related. PMID- 9406198 TI - Expression of a testis-specific putative actin-capping protein associated with the developing acrosome during rat spermiogenesis. AB - Actin-capping proteins are ubiquitous components of mammalian cells. They are known to regulate the polymerization state of actin and hence indirectly control the activity of the cytoskeleton and cell shape. As part of our investigation into the molecular mechanisms that direct differentiation of a round spermatid into an elongating spermatozoa, we report on a testis-specific 1.7-kb transcript from rat testis with sequence similarities to the alpha subunit of actin-capping proteins (ACPs) from somatic cells. The transcript contains a putative cAMP responsive motif (CREM) upstream of the initiation codon in the DNA sequence and is expressed postmeiotically, first appearing between 20 and 30 days of postnatal development. The primary amino acid sequence is 90% identical to that of a previously identified testis-specific mouse protein, gsg3, both showing approximately 40% homology to the alpha subunit of somatic ACPs. An affinity purified polyclonal antibody to a synthetic peptide derived from the rat transcript identified a 32-kDa protein on Western blots of testicular extracts. Indirect immunofluorescent localization of the protein on frozen sections of adult rat testis showed that it is intracellular and accumulates asymmetrically in the cytoplasm of round spermatids coincident with the position of the developing acrosome. This spatial expression parallels the distribution of F actin during sperm differentiation, supporting the hypothesis that testis specific ACPs have an important role in determining the final shape of mature sperm heads. A disturbance in the expression of these ACPs may underlie many of the abnormalities in sperm morphology observed in infertile semen. PMID- 9406199 TI - The metabolic properties of spermatozoa from the epididymis of the tammar wallaby, Macropus eugenii. AB - The utilization of various substrates by sperm from the cauda epididymidis of the tammar was examined because the major naturally occurring sugar in the semen of this species is N-acetyl-D-glucosamine (NAG) and not furctose, as in eutherian mammals. The sperm displayed a high level of endogenous respiration that supported motility for relatively prolonged periods of time in vitro. They also metabolised exogenous 14C-labelled glucose, NAG, sucrose, and acetate through glycolytic and/or oxidative processes to produce lactate and 14CO2 at varying rates. The rate of uptake of NAG by tammar sperm was about four times greater than that of other substrates. Glucose and/or NAG stimulated the rate of oxygen consumption by about 20%, but acetate stimulated oxygen consumption by more than 40%. The most striking findings were that NAG almost completely inhibited the oxidation of glucose and sucrose by the sperm and depressed the uptake of glucose, 3-O-methylglucose, and sucrose. Acetate oxidation also was inhibited by NAG, but only by about 50%. Tammar sperm generated substantial amounts of free glucose during incubation with NAG, but this and the inhibitory effects of NAG on glucose oxidation were not mimicked by rat sperm. It is proposed that tammar sperm fail to oxidise glucose in the presence of NAG because of the rapid cellular uptake of NAG relative to glucose. Also, the intracellular glucose and acetate liberated from NAG would compete with exogenous glucose for processing in the Embden-meyerhof and tricarboxylic acid (TCA) cycle pathways. It is also suggested that tammar sperm oxidise sucrose after extracellular hydrolysis into its glucose and fructose components. The biological implications of these metabolic and transport properties of tammar sperm have as yet to be determined. PMID- 9406200 TI - MR angiography of the supra-aortic arteries using a dedicated head and neck coil: image quality and assessment of stenoses. AB - Our purpose was to evaluate a dedicated head and neck coil for demonstration of supra-aortic arteries with optimised magnetic resonance angiography techniques. We performed 47 examinations with a 1.5-T system. We used coronal 3D fast imaging with steady precession (FISP), axial 3D tilted optimised nonsaturating excitation (TONE) and 2D fast low-angle shot (FLASH) for the carotid bifurcation, axial 3D TONE with or without magnetisation transfer (MT) for intracranial arteries, and axial 3D FISP or TONE for the aortic arch. Evaluation included visual assessment of image quality and grading of stenoses near the carotid bifurcation; digital subtraction angiography was used as the reference method. Axial 3D TONE gave superior image quality at the carotid bifurcation, MT-TONE intracranially, and 3D FISP for the aortic arch vessels. Nevertheless, sensitivity and specificity for detection of significant stenoses were similar with coronal 3D FISP (96.3%, 94.0%), axial 3D TONE (92.6%, 92.5%) and axial 2D FLASH (96.3%, 86.6%). Image quality at the aortic arch needs further improvement. PMID- 9406201 TI - False-negative angiograms in subarachnoid haemorrhage due to intracranial aneurysms. AB - Of 440 patients with spontaneous subarachnoid haemorrhage in whom an aneurysm was suspected, 60 had a negative angiogram. A second angiogram performed 1-4 weeks later revealed an aneurysm in 5 of 40 cases. Of these patients, 3 had a second haemorrhage. In all cases, diffuse bleeding, with involvement of the anteroinferior interhemispheric fissure, was present on CT. There were three aneurysms of the anterior communicating artery and two of the carotid siphon. The reasons for the false-negative angiograms and the usefulness of repeated angiography are discussed. PMID- 9406202 TI - Thrombosis of the deep cerebral veins: CT and MRI findings with pathologic correlation. AB - Deep cerebral vein thrombosis can present with acute, severe neurological symptoms and may be rapidly fatal as in the 20-year-old woman reported here. Although MRI is superior for establishing the diagnosis, CT is usually the first examination performed in the clinical setting. It is therefore important to recognise certain indicators such as extensive bithalamic low density. These and certain other less specific signs are correlated with the MRI and autopsy findings. PMID- 9406203 TI - MELAS presenting as migraine complicated by stroke: case report. AB - A case of MELAS (mitochondrial encephalomyopathy, lactic acidosis, and stroke like episodes) which presented as migraine complicated by stroke is reported. Strokes associated with migraine have often been reported, but the mechanism remains unclear and may include a variety of pathologies. MELAS also presents with migrainous headache, vomiting, and stroke-like symptoms. Magnetic resonance imaging demonstrates characteristic findings. MELAS should be considered in the differential diagnosis of infarct-like lesions with migrainous headaches in young adults, especially if the symptoms fluctuate and are accompanied by a homonymous hemianopia. PMID- 9406204 TI - MRI "fogging" in cerebellar ischaemia: case report. AB - Subacute cerebral infarcts may appear normal on T2-weighted MRI as an area isointense with surrounding normal tissue. This MRI "fogging effect" has been described in only a few cases. We present a further case of fogging observed during the evolution of a cerebellar infarct. PMID- 9406205 TI - Temporal lobe epilepsy with varying severity: MRI study of 222 patients. AB - MRI was performed in 222 consecutive adult patients with temporal lobe epilepsy of varying severity from January 1991 to May 1993. The diagnosis of hippocampal sclerosis was established visually by three independent observers. The accuracy of visual assessment of hippocampal asymmetry was compared with volumetric measurements. Neuropathological correlations were obtained in 63 patients with refractory seizures. Temporal lobe abnormalities were observed in 180 patients (81%) as follows: hippocampal sclerosis in 122 (55%); developmental abnormalities in 16 (7.2%); tumours in 15 (6.8%); scars in 11 (5%); cavernous angiomas in 10 (4.5%); miscellaneous lesions in 6. MRI was normal or showed unrelated changes in 42 patients (19%). Visual assessment correctly lateralised hippocampal sclerosis in 79 of the 84 patients measured (94%). Temporal lobectomy confirmed the MRI data (side and aetiology) in all 63 operated patients. Patients with normal MRI had an older age of seizure onset and were more often drug-responsive than patients with hippocampal sclerosis. MRI showed temporal lobe abnormalities in 81% of epileptic patients with varying severity with good neuropathological correlation. Patients with normal MRI had a less severe form of the disease. PMID- 9406206 TI - Semi-automated extraction of brain contours from MRI. AB - We compared brain volumes computed by trained operators using the BRAIN-MAP algorithm, which automatically extracts the contours of the brain from gradient echo magnetic resonance images. The images of 19 subjects randomly selected from a group of normals and a group of patients with dementia were included. BRAIN-MAP found brain perimeters that were on the average ca. 3% tighter than those obtained by two experienced operators. Between-operator and within-operator reproducibility of the analyses were also estimated and found to be (-0.13 +/- 0.51)% and (-0.63 +/- 0.08)%, respectively. Replicate volume measurement by the computer alone provided a reproducibility of (0.44 +/- 0.46)%. PMID- 9406209 TI - MRI of the anterior optic pathways following enucleation. AB - We examined five patients who had enucleation of one eye for inflammatory or neoplastic disease, using MRI at 1.5 Tesla. None had symptoms referable to the enucleated orbit. In addition, age- and-sex matched individuals were imaged as control subjects, and a further 15 subjects, referred for other than orbital disease, were reviewed. Measurements were made retrospectively of the dimensions of the optic chiasm to establish normal values. All five patients showed abnormalities on MRI following enucleation: abnormal signal within the optic nerve remnant on short tau inversion recovery (STIR) images, and atrophy of the nerve remnant and the chiasm. These findings were not apparent in the control or normal subjects. Such findings are to be expected following enucleation and should not be interpreted as indicating active pathology. PMID- 9406207 TI - Histologically confirmed changes on CT of reoperated low-grade astrocytomas. AB - During a 15-year period 37 patients with primary low-grade astrocytoma have been operated upon twice in our institute. CT and histological data at the time of the first and second operations were compared. The majority of primary astrocytomas showed as a low-density area without contrast enhancement; minimal, mainly marginal enhancement was present in six cases. At reoperation 10 tumours were histologically unchanged; the corresponding CT studies displayed a nonenhancing lesion, although insignificant contrast uptake could be seen in three cases. There were 18 tumours which had transformed into anaplastic astrocytoma: CT before repeat surgery showed more or less marked enhancement. In all nine cases which progressed into glioblastoma multiforme strong contrast enhancement was seen on CT at the time of recurrence. Although the grade of contrast uptake varied, the appearance of or increase in enhancement was a sign of some degree of anaplastic change, particularly convincing in cases of dedifferentiated glioblastoma multiforme. PMID- 9406208 TI - Intracranial meningeal melanocytoma: CT and MRI. AB - We report the MRI and CT findings of an intracranial meningeal melanocytoma (IMM) arising from Meckel's cave and review the imaging characteristics of IMM. On CT, IMM constantly appear as well-circumscribed, isodense to slightly dense, extra axial tumours with homogeneous contrast enhancement. This appearance is non specific and similar to that of meningiomas or small neuromas. On MRI, the signal of IMM is strongly related to the amount of melanin pigment: the more melanin, the more shortening of T1 and T2 relaxation times. Only when it shows as a homogeneous mass, bright on T1 and dark on T2 weighting, can a specific diagnosis of a melanin-containing tumour be made. However, this still cannot provide a distinction between IMM and malignant meningeal melanoma. PMID- 9406210 TI - Positional MRI: a technique for confirming the site of leakage in cerebrospinal fluid rhinorrhoea. AB - We report confirmation of the site of leakage in two patients with spontaneous cerebrospinal fluid (CSF) rhinorrhoea by demonstrating CSF leaking on MRI. Both patients had midline anterior cranial fossa floor (cribriform plate/fovea ethmoidalis) dural-bone defects with arachnoid herniation with or without brain herniation into the upper part of the nasal cavity on MRI, which was subsequently confirmed surgically. Corresponding to the history of postural induction or aggravation of the rhinorrhoea, the CSF leak was demonstrated by the appearance of or increase in the sinonasal fluid collection by imaging the patient in the position of maximum leakage following initial images in the supine position. PMID- 9406211 TI - Osteoid osteoma of the petrous bone. AB - We present a case of osteoid osteoma of the petrous bone presenting with progressive sensorineural hearing loss. CT showed a dense homogeneous mass at the promontory surrounded by a thin bony border. On MRI this lesion gave intermediate signal intensity on T1- and T2-weighted spin-echo images and enhanced intensely with gadolinium. Surgical removal and pathological study proved the diagnosis. PMID- 9406212 TI - "Eraseroma" as a cause of rhinolith: CT and MRI in a child. AB - We report a 5-year old girl with progressive difficulty in breathing through the nose whose clinical diagnosis was nasal tumour. CT showed a calcified nodular mass and MRI a nonspecific nodular lesion in the right nasal cavity. The radiological suspicion was a rhinolith. The operative specimen showed that an eraser from a pencil was the primary source. We underline the rarity of this entity and the important role of radiological studies in preoperative recognition. PMID- 9406213 TI - Basis for a method of dynamic proprioceptive correction in the restorative treatment of patients with residual-stage infantile cerebral palsy. AB - A new method for the restorative treatment of patients with residual-stage infantile cerebral palsy is described; the method is based on proprioceptive correction using an "Adeli-92" device, which is a modified space suit used in weightless conditions. The "Adeli-92" allows intensification and some extent of normalization of afferent proprioceptive mobility-controlling input. Positive clinical effects were obtained in 70% of patients, with improvements in walking and self-care ability. The positive effects of this method were demonstrated objectively using electroencephalography, electroneuromyography, studies of somatosensory evoked potentials, and studies of the vestibular system. PMID- 9406214 TI - New approaches to the rehabilitation of patients with neurological movement defects. AB - Results were obtained using a new method, based on the "Adeli-92" therapeutic space suit, for the rehabilitation of patients with movement disorders due to acute lesions of the cerebral circulation, head trauma, and other causes. A variety of methods was used to assess the state of patients before and after treatment, including clinical studies, psychological tests, EEG recordings, evoked potential studies, stabilography, and heart rhythm analysis. The results obtained demonstrate the high efficacy of this new method. PMID- 9406215 TI - Functional state of segmental motoneurons in Wilson-Konovalov hepatocerebral dystrophy. AB - Stimulatory electromyography was used to investigate 12 patients, and chronaximetric investigations were performed in 18. Peak H-potential amplitudes indicated reductions in reflex excitability in the motoneuron pool. The stimulus size required for inducing threshold H-reflexes demonstrated a high level of reflex excitability among those alpha-motoneurons probably involved in generating hyperkinesia. Motor response parameters showed the direct electrical excitability of motor units to be elevated. Changes in H-reflex and M-response parameters demonstrated functional rearrangement of the movement analyzer after stereotaxic surgery. The functional states of the alpha- and gamma-systems are discussed. PMID- 9406216 TI - Corticostriatal mechanisms of behavior. AB - This article presents the results of three series of experiments on cats, dogs, and lower primates, performed to investigate the structural, neurophysiological, and mediator mechanisms of the corticostriatal systems involved in the organization of behavior. Morphological studies of corticostriatal connections showed that along with the diffuse distribution of afferent terminals within the striatum, there were also elements of topical organization defined by anteroposterior and mediolateral gradients. Neurophysiological experiments on dogs and lower primates were used to study the spike activity of the prefrontal region of the cortex and the head of the caudate nucleus during training to conditioned first- and second-order reflexes and during the solution of complex problems involving delayed spatial selection. Studies demonstrated that while in dogs, most of the neurons recorded showed a transition to responses to the conditioned signal at a particular stage of carrying out a conditioned response, in monkeys all cells recorded showed specific responses at different periods of solving the task at all stages of the study. Neuropharmacological experiments on dogs showed that agents blocking glutamine receptors in the caudate nucleus had more pronounced effects at the phase of developing conditioned movement reflexes. Administration of these agents during the reflex reinforcement phase affected only the differentiation of inhibition. These results lead to the conclusion that the prefrontal area of the cortex and, to some extent, the caudate nuclei, act on incoming information specifying the current dominant need and the states of the external and internal environments, to carry out programmed actions and assess the results of these actions. PMID- 9406217 TI - Comparative study of the roles of ACTH and beta-endorphin in regulating conditioned reflex activity in the hedgehog. AB - Data on the relative effects of the neurohormone ACTH1-39 and the opioid peptide beta-endorphin on conditioned reflex activity in the hedgehog are presented. It was demonstrated that administration of ACTH (30-50 micrograms/kg s.c.) led to facilitation of learning and strengthening of memory processes (conditioned reflex traces). ACTH promoted strengthening of movement, orientational investigative, and intersignal activities, produced hyperalgesia, and blocked the effects of naloxone. Administration of beta-endorphin (30-40 micrograms/kg s.c.) lengthened the latent periods of conditioned reflexes, produced a pronounced analgesic effect, and reduced movement and intersignal activities. The effects of beta-endorphin were eliminated by dosage with naloxone. Administration of beta endorphin blocked the inhibitor effects of stimulation of the limbic cortex; doses of ACTH produced partial release of inhibitory effects. The differences between the effects of ACTH and beta-endorphin on higher nervous activity are discussed, as are the possible mechanisms of these effects. PMID- 9406218 TI - Effect of veratridine on the release of neurotrophic factors in nerve tissue cultures. AB - The depolarizing agent veratridine was shown to affect the level of neurotrophic substances in combined cultures of neonatal rat hippocampus and chick embryo spinal ganglia. In this experimental model, the level of neurotrophic factors in rat hippocampus explants increased as a result of increases in neuronal activity mediated by veratridine. The effects of these neurotrophins on neurite growth in the sensitive spinal ganglion neurons in the combined cultures were evaluated using morphometric methods. Neurite-stimulating effects were seen when veratridine was added to the nutritive medium at a concentration of 90 nM. Antibody to nerve growth factor blocked the action of veratridine. These results demonstrate a role for neuron activity as a regulatory mechanism controlling the expression of neurotrophins. PMID- 9406219 TI - Synaptic responses of neurons in the parietal associative cortex of the cat to stimulation of the red nucleus. AB - Acute experiments on anesthetized and immobilized cats using intracellular recording were used to study the responses of neurons in the parietal associative cortex to stimulation of the red nucleus. Efferent neurons of the parietal cortex were identified by antidromal activation on stimulation of the intrinsic nuclei of the pons and motor cortex. Oligo- and polysynaptic EPSP in response to stimulation of the red nucleus were seen. The results are discussed in the light of the morphological organization of the rubrothalamic and cerebellothalamocortical tracts. PMID- 9406220 TI - Quantitative characteristics of changes in synaptic contacts in the hippocampus in Alzheimer's disease. PMID- 9406221 TI - Afferent projections of the body of the caudate nucleus in the cat brain. PMID- 9406222 TI - Spatial organization of hypothalamic neurons projecting to the "gastric" region of the vagosolitary complex. PMID- 9406223 TI - Adrenal cortex hormones and the testicles in emotional stress in rats with a genetic predisposition to cataleptic responses. PMID- 9406224 TI - Antidromal and synaptic activation of neurons of the associative parietal cortex of the cat brain elicited by spike activity from the intrinsic nuclei of the pons. AB - Acute experiments were performed on cats with intracellular recording of efferent and unidentified neurons of the anterior suprasylvian and posterior lateral gyri of the parietal cortex, to study the antidromal and synaptic responses to stimulation of the lateral and medial groups of intrinsic nuclei of the pons. Oligo- and polysynaptic components were detected, along with complex EPSP due to convergence of axons from fast- and slow-conducting neurons. Antidromal and synaptic responses were demonstrated in the same parietal cortex neurons, demonstrating a double connection between the intrinsic nuclei of the pons and the associated parietal cortex. The possible pathways of these connections are discussed, along with their features and importance in the functioning of pontocortical connections. PMID- 9406225 TI - Use of artificial ion channels for quasi-intracellular recording of cerebral cortex neuron activity. AB - Action potentials and synaptic potentials were recorded in vivo from cortical neurons in baby rats aged 20-25 days using a new method based on the ionophore nystatin. Nystatin solution was used to fill a standard glass extracellular microelectrode, and became inserted into membranes. Spikes which were initially recorded as extracellular spikes showed increases in amplitude and were transformed into unipolar quasi-intracellular spikes at 0.5-5.0 min after formation of the high-conductance contact. This method allows stable recording of neuronal activity from cells for at least 1 h, and provides a good signal-to noise ratio. The electrode does not puncture the neuron membrane, with the result that experiments do not require any isolation from vibration. Thus, the results obtained demonstrate that the method is highly efficient for recording the in vivo activity of small nerve cells. PMID- 9406226 TI - Responses of constantly firing motor units to afferent stimulation. AB - Constantly firing motor units of the short abductor muscles of the first finger in the human hand and the abdominal wall muscle in immobilized rats responded to afferent stimulation of the median and sciatic nerves respectively with changes in the nature of spike activity. In the first 250 msec of the post-stimulus period, the frequency of motor unit spike activity became unstable and peri stimulus histograms were individually quite distinct. This was followed by relative stabilization of motor unit discharge frequencies, and the subsequently (750 msec) determined motor unit spike frequency depended on most cases on the background spike frequency. PMID- 9406227 TI - Studies of the mechanism of the anticonvulsant effect of delta-sleep-inducing peptide in conditions of increased oxygen tension. AB - Studies of the protective actions of three doses of delta-sleep-inducing peptide (DSIP) given at different times before barochamber compression of animals to an oxygen tension of 0.7 MPa showed that the optimum DSIP dose is 12 micrograms/100 g. Intraperitoneal administration of this dose of DSIP delayed the onset of generalized convulsive activity by a factor of 2-2.5 in animals exposed to an oxygen tension of 0.7 MPa and promoted normalization of the sleep-walking cycle within 24 h after exposure to hyperbaric oxygen (HBO), by creating an optimal balance between excitatory and inhibitory amino acid neuromediators. PMID- 9406229 TI - Monoamine metabolism in the striatum of the rat brain during drug infusion into the nucleus accumbens. AB - Studies on conscious Sprague-Dawley rats using intracerebral dialysis in live animals combined with high-performance liquid chromatography with electrochemical detection showed that administration of apomorphine into the nucleus accumbens decreased the levels of dihydroxyphenylacetic acid and 5-hydroxyindoleacetic acid in the extracellular space of the dorsal striatum throughout the observation period and produced a transient reduction in the level of homovanillic acid in the dialysate from this structure. The studies demonstrated that reversible exclusion of the nucleus accumbens with procaine produced a transient increase in the levels of dopamine metabolites, without an increase in serotonin metabolites, in the extracellular space of the dorsal striatum. These results demonstrate that the nucleus accumbens affects dopamine metabolism in the striatum, this being mediated by the dopamine-reactive system in the nucleus accumbens. PMID- 9406228 TI - Effects of the cholinergic system of the rat neostriatum on learning active escape in normal animals and in animals with lesions to the intralaminar thalamic nuclei. AB - Studies were carried out into the role of the parafascicular (Pf) nuclei of the rat thalamus in learning a conditioned active escape reflex (CAER) in a T-maze, a reflex associated with discrimination of visual stimuli, and into the regulatory effect on this learning process of activation of the neostriatal cholinergic system. The following results were obtained using 57 Sprague-Dawley rats divided into a number of experimental groups: 1) bilateral microinjection of carbacholine (0.03 microgram) into the neostriatum on days 4, 5, and 6 of training produced significant (p < 0.01) increases in the proportion of correct discriminant CAER performances; 2) bilateral lesioning of the Pf nuclei led to irreversible disruption of the previously learned CAER. Rats with initially bilaterally lesioned Pf nuclei did not learn the discriminant CAER at all after 10 days of training (16 combinations), and microinjection of carbacholine into the neostriatum of these animals was ineffective. It is concluded that the integrity of the afferent input into the Pf nuclei of the thalamus is an important factor for activation of the neuronal background of the neostriatum, and is required for cholinergic activation of the neostriatum to be effective. PMID- 9406230 TI - Comparative physiological features of the regulatory effect of vasopressin on higher nervous activity in an ascending series of mammals. AB - This report provides comparative physiological data on the features of the regulatory effects of the neurohormone vasopressin on higher nervous activity in an ascending series of mammals consisting of insectivores, rodents, and primates. Administration of vasopressin to hedgehogs produced a general facilitatory effect on conditioned reflex brain activity. The effects of vasopressin on memory processes in hedgehogs was minor. In rabbits, vasopressin had greater regulatory effects on conditioned reflex memory than in hedgehogs. However, this was transient in nature. In monkeys, administration of vasopressin had complex differential effects on simple conditioned responses and different types of memory. The effects of vasopressin on memory processes were long-lasting and were different for corticalized and noncorticalized forms of nervous activity. The question of changes in the nature of the regulatory effects of vasopressin during phylogenesis is discussed, as is the question of the increases in its level of involvement in the regulation of higher nervous functions and memory processes. PMID- 9406231 TI - Proteolytic cleavage of p53: a model for the activation of p53 in response to DNA damage. AB - p53 is a multifunctional protein that reacts to DNA damage within the cell and regulates the cell growth arrest and/ or apoptotic pathways. However, the mechanism of p53 activation in response to DNA damage is unknown. Recently we have shown that interaction of p53 with sites of DNA damage induces selective proteolytic cleavage of p53, resulting in fragments of 40 and 35 kDa molecular weight. We have also shown that interaction of p53 with single-stranded (ss)DNAs results in a different pattern of selective proteolysis. This interaction gives a novel of 50-kDa protein generated by C-terminal cleavage of the full length protein and released from the p53-ssDNA complexes. Here we discuss a model where p53 responds to the DNA damage by generating different sets of the proteolytic fragments according to the type of the damage. PMID- 9406232 TI - The structure and functions of the HAP1/Ref-1 protein. AB - The HAP1/Ref-1 (hereafter referred to as HAP1) protein is a nuclear enzyme that apparently performs two distinct roles in the cellular defense against oxidative stress. One well-established role is in the repair of a variety of lesions induced in DNA either by spontaneous hydrolysis or by reactive oxygen species (ROS). This function has been characterized in great detail and the roles played by individual active site amino acid residues have been defined. The second role, which was identified only relatively recently and is still not characterized in detail, is to regulate the DNA binding activity of a group of nuclear factors. This second role proceeds via the modification of the oxidation/reduction (redox) state of a cysteine residue in the target protein, although the mechanism by which this is achieved remains to be elucidated. In this article, we shall review the latest knowledge on the relationship between structure and the dual functions of HAP1, and we will seek to explain in detail the roles played by several individual amino acid residues in the catalytic function of the HAP1 protein. PMID- 9406233 TI - Modulation of antioxidant defenses during apoptosis. AB - Understanding the fundamental mechanism of apoptosis is crucial to developing therapeutic strategies for controlling apoptosis in diseased tissues. We are using model systems with relevance to cancer treatment to investigate the mechanism of apoptosis. Subtraction hybridization cloning was used to identify transcripts present at higher levels in regressing vs. normal prostate; these may be important for apoptosis. One of the genes cloned from regressing prostate is also upregulated in the murine W7.2 lymphocyte cell line induced to undergo apoptosis by treatment with the synthetic glucocorticoid, dexamethasone. This gene encodes a mu class glutathione S-transferase (EC 2.5.1.18), a protein that can protect the cell against oxidative stress by repairing oxidized lipids, proteins, and DNA. Glutathione S-transferase expression does not increase with dexamethasone treatment of lymphocyte cell lines expressing nonfunctional glucocorticoid receptors or a mutation in the apoptotic pathway. Other antioxidant defenses, including catalase (EC 1.11.1.6) and superoxide dismutase (EC 1.15.1.1), decline following dexamethasone treatment of W7.2 cells. Overexpression of the bcl-2 oncogene protects these cells against dexamethasone mediated apoptosis and prevents the decrease in antioxidant enzyme activity. These findings support the hypothesis that control of the cellular redox state is important to the mechanism of glucocorticoid-mediated lymphocyte apoptosis. Another model system we are using is tumor necrosis factor-alpha treatment of MCF 7 human breast cancer cells. Our preliminary results suggest that, in this system, activation of nuclear factor-kappa B and increased expression of manganese superoxide dismutase may afford protection from apoptosis. PMID- 9406234 TI - DNA damage from oxidants: influence of lesion complexity and chromatin organization. AB - DNA damage by reactive oxygen species results in a spectrum of DNA lesions including single-strand breaks (ssb) and double-strand breaks (dsb). However, most damage is not lethal, and the location and nature of the DNA damage, in addition to total number of breaks, are likely to be critical in determining ultimate survival. Generally associated only with ionizing radiation, multiply damaged sites (i.e., complex lesions and clusters of complex lesions in DNA) are more likely to be lethal because they are less easily repaired. We examined five drugs known to cause DNA adducts, strand breaks, and reactive oxygen species for their ability to produce complex lesions: 4-nitroquinoline-1-oxide (4NQO), H2O2, doxorubicin, Tirapazamine, and etoposide. As indicators of lesion complexity we compared 1) the ratio of ssb to dsb, 2) the rate of rejoining of single-strand breaks, 3) the relative lethality of the breaks (number of breaks per mean lethal dose), and 4) the ability to produce complex lesions. Tirapazamine, etoposide, and doxorubicin gave dsb/ssb ratios similar to that for X-rays, whereas 4NQO and H2O2 showed dsb/ssb ratios of 200 and 3250, respectively. The number of dsb per LD50 varied from 2.5 to 500 for different drugs. There was no apparent relation between ssb rejoining half-time (3.5-85 min) and relative lethality or lesion complexity. A modified (nonionic detergent) filter elution method confirmed that tirapazamine, like ionizing radiation, produced multiple dsb within single chromatin domains. These data indicate that complex lesions can be produced by a number of different chemicals and suggest that the damage that results in killing by these drugs may be related to production of multiply damaged sites in DNA. PMID- 9406235 TI - Importance of glutathione and associated enzymes in drug response. AB - Maintenance of cellular homeostasis is a critical survival trait in tumors when exposed to anticancer drugs. Because conjugation and elimination of drugs and their metabolites is dependent upon sequential and coordinated pathways, acquired drug resistance through a gradual adaptive response would rarely be expected to be the consequence of changes in the expression of one gene product. We have used a number of drug-resistant human cell lines to characterize those genes that are implicated in maintaining a resistant phenotype. Human HT29 colon cancer cells chronically exposed to ethacrynic acid (EA) [a glutathione (GSH) and glutathione S-transferase (GST) modulator] have acquired resistance to the drug. Commensurate with resistance, EA is more effectively conjugated to GSH and effluxed from the resistant cells. Using directed and random (differential display) approaches, a number of detoxification and/or protective gene products have been shown to be expressed at elevated levels. These include: gamma-glutamyl cysteine synthetase (gamma-GCS, the rate-limiting enzyme in GSH biosynthesis); GST pi (the enzyme catalyzing the conjugation reaction); multidrug resistance associated protein (MRP) (the membrane pump responsible for effluxing the conjugate from the cell interior). In addition, other gene products not directly linked with EA metabolism were induced, including dihydrodiol dehydrogenase (an alpha ketoreductase) (30-fold), DT-diaphorase (threefold), and a transcriptional regulator SSP 3521 (threefold). HL60 cells resistant to a GSH paralog Ter199 also show increased expression of some of these gene products. Furthermore, an adriamycin-resistant human HL60 cell line also shows overexpression of GST pi, gamma-GCS, and MRP, but in addition has approximately 20-fold more DNA-dependent protein kinase catalytic subunit (DNA-PKcs). This enzyme is an early stress response gene that can phosphorylate and activate downstream transcription factors. Such overexpression could impact on the transcriptional control of the other detoxification gene products. Both adriamycin and a typical drug-GSH conjugate (APA-SG) are inhibitors of DNA-PK. Because cellular levels of these conjugates would presumably be a good indicator of stress, it would seem reasonable to speculate that DNA-PK may act as a receiver and transmitter of signals that are crucial to the drug-resistant phenotype. Additionally, this enzyme may prove to be a potentially important target for drug design based upon the inhibitory activity of GSH conjugates. PMID- 9406236 TI - Selenium and the thioredoxin redox system: effects on cell growth and death. AB - Thioredoxin is a redox protein found overexpressed in some human tumors. Thioredoxin is secreted by tumor cells and enhances the sensitivity of the cancer cells to other growth factors. Redox activity is essential for stimulation of cell growth by thioredoxin. Cells transfected with thioredoxin cDNA show increased tumor growth and decreased apoptosis in vivo and decreased sensitivity to apoptosis induced by a variety of agents both in vitro and in vivo. Cells transfected with a redox-inactive mutant thioredoxin show inhibited tumor growth in vivo. Dietary selenium has been shown to prevent some forms of human cancer. Selenocysteine is an essential component of thioredoxin reductase, the flavoenzyme that is responsible for the reduction of thioredoxin. Selenium added to the culture medium increases thioredoxin reductase activity due to an increase in thioredoxin reductase protein but mostly due to an increase in the specific activity of the enzyme. Some diaryl chalcogenide (selenium and tellurium) compounds have been studied as inhibitors of thioredoxin reductase. The most active were diaryl tellurium compounds, which were noncompetitive inhibitors of thioredoxin reductase with Ki values of 2-10 microM. Several of the compounds inhibited cancer cell colony formation in vitro with IC50s as low as 2 microM. PMID- 9406237 TI - Targeting gene therapy to cancer: a review. AB - In recent years the idea of using gene therapy as a modality in the treatment of diseases other than genetically inherited, monogenic disorders has taken root. This is particularly obvious in the field of oncology where currently more than 100 clinical trials have been approved worldwide. This report will summarize some of the exciting progress that has recently been made with respect to both targeting the delivery of potentially therapeutic genes to tumor sites and regulating their expression within the tumor microenvironment. In order to specifically target malignant cells while at the same time sparing normal tissue, cancer gene therapy will need to combine highly selective gene delivery with highly specific gene expression, specific gene product activity, and, possibly, specific drug activation. Although the efficient delivery of DNA to tumor sites remains a formidable task, progress has been made in recent years using both viral (retrovirus, adenovirus, adeno-associated virus) and nonviral (liposomes, gene gun, injection) methods. In this report emphasis will be placed on targeted rather than high-efficiency delivery, although those would need to be combined in the future for effective therapy. To date delivery has been targeted to tumor specific and tissue-specific antigens, such as epithelial growth factor receptor, c-kit receptor, and folate receptor, and these will be described in some detail. To increase specificity and safety of gene therapy further, the expression of the therapeutic gene needs to be tightly controlled within the target tissue. Targeted gene expression has been analyzed using tissue-specific promoters (breast-, prostate-, and melanoma-specific promoters) and disease-specific promoters (carcinoembryonic antigen, HER-2/neu, Myc-Max response elements, DF3/MUC). Alternatively, expression could be regulated externally with the use of radiation-induced promoters or tetracycline-responsive elements. Another novel possibility that will be discussed is the regulation of therapeutic gene products by tumor-specific gene splicing. Gene expression could also be targeted at conditions specific to the tumor microenvironment, such as glucose deprivation and hypoxia. We have concentrated on hypoxia-targeted gene expression and this report will discuss our progress in detail. Chronic hypoxia occurs in tissue that is more than 100-200 microns away from a functional blood supply. In solid tumors hypoxia is widespread both because cancer cells are more prolific than the invading endothelial cells that make up the blood vessels and because the newly formed blood supply is disorganized. Measurements of oxygen partial pressure in patients' tumors showed a high percentage of severe hypoxia readings (less than 2.5 mmHg), readings not seen in normal tissue. This is a major problem in the treatment of cancer, because hypoxic cells are resistant to radiotherapy and often to chemotherapy. However, severe hypoxia is also a physiological condition specific to tumors, which makes it a potentially exploitable target. We have utilized hypoxia response elements (HRE) derived from the oxygen-regulated phosphoglycerate kinase gene to control gene expression in human tumor cells in vitro and in experimental tumors. The list of genes that have been considered for use in the treatment of cancer is extensive. It includes cytokines and costimulatory cell surface molecules intended to induce an effective systemic immune response against tumor antigens that would not otherwise develop. Other inventive strategies include the use of internally expressed antibodies to target oncogenic proteins (intrabodies) and the use of antisense technology (antisense oligonucleotides, antigenes, and ribozymes). This report will concentrate more on novel genes encoding prodrug activating enzymes, so-called suicide genes (Herpes simplex virus thymidine kinase, Escherichia coli nitroreductase, E. (ABSTRACT TRUNCATED) PMID- 9406238 TI - Hypoxia response elements. AB - Hypoxia-inducible factor-1 (HIF-1) has been shown to mediate the transcriptional activation of its target genes in response to oxygen concentration, most likely via a pathway involving a specific oxygen sensor. Molecular cloning of HIF-1 has shown that this widely expressed, DNA binding transcription factor is a heterodimer of two proteins, HIF-1 alpha and HIF-1 beta. A major control of HIF-1 activity by oxygen tension is achieved by changes in the level of the HIF-1 alpha subunit, which complexes with the constitutively expressed HIF-1 beta subunit. Such changes in HIF-1 alpha abundance occur via regulated stability, probably involving proteolysis, rather than at the level of transcription or translation. Further analysis has shown the existence of two separate regulatory domains in the C-terminus of the alpha subunit. Thus, a mechanism of oxygen-regulated HIF-1 activation is proposed, which involves the operation of one inducible domain being amplified by changes in protein level conferred by a second regulatory domain. Evidence for a critical role of HIF-1 in the response of diverse target genes involved in cellular growth and metabolism comes from studies on cultured, mutant mouse cells that lack a functional HIF-1 beta subunit. Furthermore, studies on tumor xenografts derived from the mutant and wild-type cells show that HIF-1 is activated in vivo, and has major effects on gene expression in response to tumor hypoxia. Thus, HIF-1 is a critical component of the oxygen-signaling pathway, and is a prime candidate regulator molecule for the role of coordinating vascular oxygen supply with cellular growth and energy metabolism. PMID- 9406239 TI - A new cellular target for mitomycin C: a case for mitochondrial DNA. AB - Mitomycin C (MMC) is an anticancer, antibiotic that is currently used clinically against a wide variety of tumors. Nuclear DNA is regarded as the primary cellular target for mitomycin's toxicity. In this study, the effect of MMC on mitochondrial DNA was tested both in vitro and in vivo. EMT6 mouse mammary carcinoma cells were treated with MMC and conformational changes in their mitochondrial DNA were determined as a measure of mitochondrial DNA damage. A dose-dependent relationship was observed between MMC treatment dosages and mitochondrial DNA damage. Liver tissue mitochondria from Balb/c mice treated with MMC were assayed for mitochondrial integrity. Mitochondrial integrity was lowered in the MMC-treated animals. Liver tissue adenosine triphosphate (ATP) levels were also shown to be significantly decreased in these same animals. We also show that MMC can be activated by mitochondria. These studies provide strong evidence that mitochondrial DNA is a target for MMC and that this interaction has a biochemical consequence that could prove toxic. PMID- 9406240 TI - DT-diaphorase: possible roles in cancer chemotherapy and carcinogenesis. AB - DT-diaphorase [also called NAD(P)H:menadione oxidoreductase; NAD(P)H:(quinone acceptor) oxidoreductase; quinone reductase, azo-dye reductase] (EC.1.6.99.2) has been shown to have a role in activation of quinone-containing cancer chemotherapeutic prodrugs to their active form, as well as inactivation of a variety of xenobiotics involved in carcinogenesis. To illustrate the basis of this dichotomy, a brief review of the historical and recent background of studies on DT-diaphorase is given. Recent data from the laboratory of the authors on the nature of the protein coded for by a recently identified human polymorphism, a C to T transition at nucleotide 609 of DT-diaphorase cDNA, and the frequency of this mutation in anal canal carcinoma patients is presented. Finally, a series of questions about the role of DT-diaphorase in both normal and malignant cells is raised based on the results of others that have been published in the last 2-3 years. PMID- 9406241 TI - Redox active disulfides: the thioredoxin system as a drug target. AB - Thioredoxin, and particularly extracellular thioredoxin, presents an attractive target for developing novel agents to treat cancer. Our studies have involved the examination of a series of alkyl 2-imidazolyl disulfides as inhibitors of the growth-stimulatory activity of the thioredoxin system. We originally determined the disulfides to be weak reversible inhibitors of thioredoxin reductase. Subsequently, we have shown that alkyl 2-imidazolyl disulfides interact directly with thioredoxin, thioalkylating critical cysteine residues or causing dimerization of the protein leading to its loss of biological activity. One of the analogues that binds to thioredoxin, 1-methylpropyl 2-imidazolyl disulfide (IV-2), selectively inhibits the thioredoxin-dependent growth of tumor cells in culture and has antitumor activity against MCF-7 and HL-60 tumors in vivo. Our work involves the development of a parallel combinatorial synthetic method to produce a large number of disulfide analogues at one time. These analogues, which differ sterically, electronically, and physically, were produced in a 96-well plate. The biological activity of these analogues was evaluated, also in the 96 well plate format. This rapid method of evaluating biological activity is a means to identify agents with specificity for inhibition of the thioredoxin system, and may provide novel antitumor agents with activity against solid tumor cancers. PMID- 9406242 TI - Nitro reduction as an electronic switch for bioreductive drug activation. AB - It is well known that the reduction of aromatic nitro groups can give rise to toxic species, and that net nitro reduction by one-electron reductases can usually be inhibited by oxygen. There has been much interest in utilizing this biotransformation to activate drugs in hypoxic regions of tumors, but no clinically useful compound has yet resulted. Nitroreductive activation of prodrugs by oxygen-insensitive (and oxygen-sensitive) reductases is also of current interest because of new methods for introducing specific nitroreductases into tumors (e.g., as antibody-enzyme conjugates or by gene therapy). In most of the compounds investigated previously, cytotoxicity appears to be due to reactive nitroso or hydroxylamine reduction products arising from the nitro group itself. It is argued that there is greater scope for designing potent and selective nitro compounds by using the nitro group as an electronic switch to activate a latent reactive moiety elsewhere in the molecule. Examples of this approach include the nitro(hetero)aromatic mustards (e.g., SN 23816, NSC 646394) in which the nitro group controls the reactivity of a nitrogen mustard to which it is directly conjugated, and the nitro(hetero)aromatic methylquaternary (NMQ) mustards (e.g., SN 25341, NSC 658926) in which reduction of the nitro group triggers fragmentation of the molecule to release a reactive aliphatic nitrogen mustard. Many of these compounds show very high selectivity for hypoxic cells in culture. Some are also active against hypoxic cells in tumors, and provide large tumor growth delays when combined with tumor blood flow inhibitors such as 5,6 dimethylxanthenone-4-acetic acid (DMXAA). These prodrug designs also have potential for releasing effectors other than nitrogen mustards, which opens up many possibilities for use of nitro compounds as tumor-selective prodrugs. PMID- 9406244 TI - Oxygen delivery: implications for the biology and therapy of solid tumors. AB - Solid malignancies develop areas of hypoxia during the earliest phase of their development, while they are still microscopic in size and before they have initiated angiogenesis. The physiologic effects of hypoxia and the associated microenvironmental inadequacies increase mutation rates, select for cells deficient in normal pathways of programmed cell death, and contribute to the development of an increasingly invasive, metastatic phenotype. Hypoxic tumor cells are also resistant to radiation and to many antineoplastic drugs, and can limit the curability of solid tumors by radiotherapy and chemotherapy. This article reviews several approaches to improving tumor oxygenation and discusses the results of laboratory and clinical studies examining their efficacy in improving the outcome of radiation therapy. Past clinical trials, using nonoptimal agents and regimens to circumvent the protective effects of hypoxia, improved local control rates by approximately 10%. Greater improvements should be possible when the new agents now undergoing laboratory testing are used in optimally designed clinical regimens. PMID- 9406243 TI - Induction of DT-diaphorase in cancer chemoprevention and chemotherapy. AB - DT-diaphorase (EC 1.6.99.2) is a flavoprotein that catalyses two-electron reduction of quinones, quinone imines, and nitrogen oxides. It is a Phase II detoxifying enzyme that can detoxify chemically reactive metabolites, and may be important in an early cellular defense against tumorigenesis. DT-diaphorase is also an activating enzyme for bioreductive antitumor agents like mitomycin C (MMC) and EO9. DT-diaphorase is induced in many tissues by a wide variety of compounds including dithiolethiones and isothiocyanates. Dithiolethiones are chemoprotective agents against a variety of chemical carcinogens in animal models, and the dithiolethione analogue, oltipraz, is currently in Phase I and Phase II clinical chemoprevention trials. Similarly, the isothiocyanate derivative, sulforaphane, blocks the formation of carcinogen-induced mammary tumors in rats. The low toxicity of these inducers of DT-diaphorase makes them suitable for use as chemopreventive agents in high-risk individuals. Cells with elevated DT-diaphorase levels are generally more sensitive to bioreductive antitumor agents. Thus, we suggested that the antitumor efficacy of bioreductive agents can be enhanced by selective induction of DT-diaphorase in tumor cells compared with normal cells. We showed that 1,2-dithiole-3-thione (D3T) can increase the level of DT-diaphorase activity and the cytotoxic activity of bioreductive agents in mouse lymphoma cells without increasing these activities in normal mouse marrow cells. D3T also increased DT-diaphorase activity in 24 of 33 human tumor cell lines representing nine tissue types with no obvious relationships between the tumor type, or the base level of DT-diaphorase activity, and the ability to increase enzyme activity. A series of dithiolethione analogues and dietary components were also shown to be good inducers of DT diaphorase in human tumor cells. D3T increased DT-diaphorase activity in normal human bone marrow and kidney cells but the increases were small in these cells. Combination treatment with D3T and EO9 increased cell kill in HL-60 human leukemia cells compared with EO9 alone, but had no effect on EO9 toxicity in normal human kidney cells. Similarly, D3T increased tumor cell kill by EO9 in H661 human lung cancer cells and by MMC in T47D human breast cancer cells. Thus, inducers of DT-diaphorase may play an important role in cancer chemoprevention programs and may also be useful in enhancing the antitumor efficacy of bioreductive agents. PMID- 9406245 TI - Bioreductive agents: a clinical update. AB - Bioreductive agents are drugs that must undergo reduction to form an active cytotoxic species. The existence within solid tumors of regions of hypoxia offers the possibility of using such bioreductive agents to exert tumor-specific cytotoxicity. The only drug with bioreductive properties that is in routine clinical use in mitomycin C. This is a relatively old drug that has cytotoxicity independent of its bioreductive properties. However, the results of recent randomized clinical trials of mitomycin C in combination with radiotherapy have suggested that its bioreductive properties may play an important part in its activity. EO9 is a new bioreductive agent. Phase I and II clinical trials with EO9 have failed to demonstrate any significant antitumor activity. However, the design of these studies was such that activity based on bioreductive properties may have been difficult to demonstrate. Tirapazamine is the lead compound of a novel class of bioreductive agents. It is currently undergoing extensive clinical evaluation alone, combined with radiotherapy, and in combination with other cytotoxic drugs. Although early results of these trials are encouraging, the results of randomized studies will be required before the true value of this drug can be assessed. PMID- 9406246 TI - Quantitative study of ductal breast cancer--patient targeted prognosis: an exploration of case base reasoning. AB - Current analytic methodologies allow the extraction, even from small tumor masses, of extensive information on the biologic characteristics of malignant lesions, such as tumor aggressivity, metastatic potential, drug resistance, and host interactions. Clinical practice now offers a wide range of therapeutic strategies. Information technological advances offer the opportunity to refer to very large data bases of patient anamnestic data, response to treatment and clinical outcome. There is a need to formulate therapy and prognosis for each individual case. Case based reasoning is a knowledge based methodology where the outcome for complex situations can be predicted by referring to a large data base of cases of known outcomes. The preliminary data obtained from this study suggest that case based reasoning may offer a promising approach to individual targeted prognosis. PMID- 9406247 TI - Comparison of standardized immunohistochemical and biochemical assays for estrogen and progesterone receptors in breast carcinoma. AB - The purpose of this study was to determine estrogen receptor (ER) and progesterone receptor (PR) expression assessed by dextran-coated charcoal method (DCC) and standardized immunohistochemical method (IHC) in a prospective series of 557 primary breast carcinomas, to assess the concordance between the two assays, and to evaluate the association between hormone receptor expression and various clinicopathological parameters. For ER, results of both methods were in agreement in 73.6% of the cases (277 positive, 133 negative), 74 tumors (13.3%) where IHC+/DCC- and 73 (13.1%) were IHC-/DCC+. For PR, concordant results were observed in 72.7% of the cases (201 positive and 204 negative), 127 tumors (22.8%) were IHC+/DCC- and 25 (4.5%) were IHC-/DCC+. Irrespective of the method used, ER and PR positivity showed a strong negative association with tumor grade. ER+ tumors were significantly more common among older patients. With IHC, PR+ cases were more common among tumors of lobular and mucinous type and among node positive tumors. The only parameter that was related to the concordance rate of ER determination by the DCC and IHC method was the age of the patients, with agreement being significantly lower in the group of patients younger than 50 years. On the other hand, discordant PR determination was more often observed in tumors of lower grade, node positive tumors and in tumors of lobular and mucinous type. PMID- 9406248 TI - Prognostic significance of Ki-67 expression in transitional cell bladder carcinoma after radical cystectomy. AB - We evaluated the prognostic significance of the Ki-67 labeling index (Ki-67 LI) in 75 patients with transitional cell carcinoma of the bladder who underwent radical cystectomy. Immunohistochemical staining of archival material was performed by the streptavidin-biotin method. Univariate survival analysis showed that Ki-67 LI (p < 0.001), histologic grade (p < 0.05), tumor stage (p < 0.001) and the number of positive lymph nodes (p < 0.001) significantly correlated with prognosis. Multivariate survival analysis indicated that the Ki-67 LI (p < 0.05), histologic grade (p < 0.01), tumor stage (p < 0.01), presence of lymph node metastases (p < 0.05) and use of neo-adjuvant therapy (p < 0.05) had independent prognostic value. The Ki-67 LI is an independent prognostic factor for patients with transitional cell bladder cancer treated by radical cystectomy. PMID- 9406249 TI - Flow cytometrical and genotypic analysis of primary non-Hodgkin's lymphoma of bone. AB - Primary malignant lymphoma of bone presents both diagnostic and therapeutic problems. No previous study has addressed the use of flow cytometry in the immunophenotypic evaluations of bone lymphomas. A 41-year-old Japanese female presented with a large lytic lesion in the right femur. Biopsies from the lesion were examined by light microscopy, immunohistochemistry, 3-color flow cytometry and Southern blotting. Microscopic examination showed a diffuse, noncleaved large cell lymphoma. The lymphoid cells were positive for mature B cell antigens and the phenotype was: CD5-, CD10-, CD19+, 20+, CD22+, IgG+, IgA-, IgM-, kappa-, lambda+, CD1-, CD2-, CD3-, CD4-, CD7-, CD8-. Southern blot analysis revealed rearranged bands on both heavy- and lambda light-chain genes, in contrast to germline configuration on T cell antigen receptor (beta, gamma and delta) genes. Flow cytometry, in conjunction with morphologic and other molecular techniques, can provide a rapid and accurate means of diagnosing primary lymphoma of bone. In addition, the capabilities of flow cytometry to assess cell properties/constituents can be utilized in detailed analysis of adhesion molecules and activation antigen which may lead to better prediction of the prognosis of this group of lymphomas, and may provide further important data for the therapeutic decision making process. PMID- 9406250 TI - Systemic lupus erythematosus (SLE) lymphadenopathy presenting with histopathologic features of Castleman' disease: a clinicopathologic study of five cases. AB - Lymph node enlargement is common in active systemic lupus erythematosus (SLE), a disease characterized by well defined clinical criteria. Although numerous reports have described the characteristic histology of SLE lymphadenopathy to include necrotizing lesions and hematoxylin bodies, no detailed description has examined the histopathologic features that are similar to Castleman's disease (CD) in SLE patients. In this report, we describe the clinicopathologic findings of CD-like peripheral lymphadenopathy, which was identified in five (26%) of 19 SLE patients. These five patients were all female with an age range of 24 to 44 years, and four of them presented with multicentric lymphadenopathy. They also had systemic symptoms and abnormal laboratory findings, indicating active disease, although two patients had not fulfilled the diagnostic criteria of SLE at the initial disease. The size of the enlarged lymph nodes seldom exceeded 2.0 cm in diameter, and biopsies revealed histopathologic features similar to CD, of intermediate type in three patients and hyaline vascular type in two according to the classification of Flendrig [7]. Immunohistochemical studies demonstrated polyclonal plasma cell populations in all five cases. Epstein-Barr virus genomes were detected in the small lymphocytes of two of the three cases examined by in situ hybridization studies. Recently, the histopathologic findings of CD have been associated with a disrupted immune response, and the present data suggest that SLE should be listed as one of the diseases showing the histopathologic features similar to CD. PMID- 9406251 TI - Hashimoto's thyroiditis associated with Riedel's thyroiditis and retroperitoneal fibrosis. AB - This article describe the case of a 38-year-old woman with simultaneous involvement of the thyroid by Hashimoto's thyroiditis and Riedels's disease, associated with retroperitoneal fibrosis and lipidic endarteritis. According to a large review of the literature on the occurrence of this rare condition the difficulty in sharply defining the two thyroid processes is discussed. The chronology of the events is analyzed, and etiopathogenic hypotheses are listed with emphasis on the relationship between the vascular lesions and the onset of the fibrosing process. PMID- 9406252 TI - A 19-week-old fetus with craniosynostosis, renal agenesis and gastroschisis: case report and differential diagnosis. AB - A case of a fetus affected with craniosynostosis, unilateral renal agenesis and gastroschisis is reported. The propositus was delivered on the 19th week of gestation for premature rupture of the membranes. Macroscopy showed turricephaly, shallow orbits, exophthalmos, hypertelorism, hypoplastic maxilla with relative mandibular prognathism and gastroschisis. Additional autopsy findings included a premature bilateral closure of the lambdoid suture and a unilateral renal agenesis. The nosological aspects of this fetus and the differential diagnosis of well-described craniosynostosis syndromes with characteristic craniofacial growth patterns (Crouzon syndrome, Jackson-Weiss syndrome, Apert syndrome, Saethre Chotzen syndrome, Pfeiffer syndrome) are discussed. PMID- 9406253 TI - Histiocytoid cardiomyopathy: a cause of sudden death in infancy. AB - We report the case of an infant aged of 14 months deceased of sudden death. The diagnosis of histiocytoid cardiomyopathy was made on a necropsic basis. The pathologic examination showed a cardiac hypertrophy characterized by yellowish areas with irregular outlines, disseminated in the myocardium, and made of histiocyte-like cells with foamy or granular cytoplasm. These cells reacted positively with desmin and myoglobin labels, and had rare and disorganised myofibrils in electron microscopy, proving their muscular origin. The illness affects infants and usually causes severe cardiac troubles leading to death without treatment. This case is the fourteenth associated with sudden death. PMID- 9406254 TI - Potential dangers in the psychiatric manpower war. PMID- 9406255 TI - Areas of practice of social workers in mental health. PMID- 9406256 TI - Restructuring mental health culture through client "commoditization". PMID- 9406257 TI - A foolproof method of quality assurance in the psychiatric emergency service. PMID- 9406258 TI - Therapists can be harmful. PMID- 9406259 TI - NARSAD: a decade of support for psychiatric research. Interview by John A Talbott. AB - The National Alliance for Research on Schizophrenia and Depression (NARSAD) celebrated its tenth anniversary this year. The organization raises and distributes funds for scientific research on brain disorders. As a member of NARSAD's scientific council since the organization was founded, I have been greatly impressed by the energy and commitment of NARSAD's leaders and staff and the many individuals who volunteer as members of the board of directors and the scientific council. I asked Constance E. Lieber, president of the board of directors, and Herbert Pardes, M.D., president of the scientific council, both of whom have been deeply involved with NARSAD since its inception, to agree to an interview with Psychiatric Services. The following is an edited transcript of the interview. PMID- 9406260 TI - Use of electroconvulsive therapy in the Medicare population between 1987 and 1992. AB - OBJECTIVE: Use of electroconvulsive therapy (ECT) in the Medicare population was examined to document trends and variations in the rate of use, expenditures, and patterns of treatment. METHODS: Medicare part B enrollment and claims data were used for a 5 percent nationally representative sample of Medicare beneficiaries for calendar years 1987 through 1992. Descriptive and multivariate analyses were performed. RESULTS: Weighted results showed that nationally the number of Medicare beneficiaries treated with ECT increased from 12,000 in 1987 to 15,560 in 1992. The rate of ECT use per 10,000 Medicare beneficiaries also increased from 4.2 to 5.1. Increases in use occurred among women, whites, and the disabled population (under age 65). Males, nonwhites, and the elderly did not share in the increase. Utilization and expenditure data showed an increase in outpatient ECT and a decrease in inpatient use between 1987 and 1992. The share of Medicare part B ECT expenditures in the outpatient setting increased steadily, from 7 percent in 1987 to 16 percent in 1992. Patients averaged eight ECT treatments, ranging from 6.7 in the West to 8.3 in the Northeast. CONCLUSIONS: The findings document that after a long period of declining use in the United States, ECT use in the Medicare population increased between 1987 and 1992. The analysis also documents a shift toward increasing use of outpatient ECT. PMID- 9406261 TI - Comparative outcomes of emotionally disturbed children and adolescents in a system of services and usual care. AB - OBJECTIVE: This study compared six-month functional and symptom outcomes of children and adolescents with serious emotional disturbance who received services in an exemplary system of care with outcomes of children who received traditional care. The system of care offers a comprehensive and coordinated network of mental health and other necessary services. METHODS: The study used a randomized longitudinal experimental design. Baseline data on symptoms, functioning, and family characteristics were collected from 350 families selected from among those who sought services for children from community agencies in Stark County, Ohio. The families were randomly assigned to either the experimental group, which received services from the system of care, or the control group, which received usual care in the community. Six-month outcome measures of children's symptoms and functioning were compared for the two groups. RESULTS: Although access to care and the amount of care received increased under the system of care, no differences in clinical or functional outcomes were found between the group served in the system of care and the group who received usual care. CONCLUSIONS: The effects of systems of care are primarily limited to system-level outcomes such as access to and cost of care and do not appear to affect clinical outcomes such as functioning and symptoms. PMID- 9406262 TI - Collaboration between hospital social work and pastoral care to help families cope with serious illness and grief. AB - The authors describe an educational program for hospital chaplains based on a partnership between pastoral care and social work in a teaching hospital. The goal of the program is to train chaplains to use social work principles to maximize families' capacity to negotiate a complex health care system, to solve problems in a health crisis, and to participate constructively on the health care team. Building on the two disciplines' shared values in person-centered, process oriented approaches to care, the program introduces chaplain trainees to social work concepts of crisis intervention and family systems theory that they can apply in their interactions with families in their hospital and congregational practice. The approach taught in the program is illustrated in a case vignette describing a chaplain trainee's work with the family of a child treated for severe burns. PMID- 9406263 TI - Influence of patient and hospital factors on consumer satisfaction with inpatient mental health treatment. AB - OBJECTIVE: This study examines patient- and facility-related determinants of satisfaction with inpatient mental health services. METHODS: A random sample of veterans discharged from Department of Veterans Affairs inpatient units with primary diagnoses of a psychiatric or substance use disorder (N = 13,574) were mailed a 73-item questionnaire that addressed aspects of their recent hospital experience. Multiple regression analysis was used to evaluate the relationship between patient and hospital characteristics and both the likelihood of responding to the survey and aspects of satisfaction measured by 14 subscales. RESULTS: A total of 4,968 veterans, or 37 percent, mailed back responses to the questionnaire. Respondents were older than nonrespondents and were more likely to be white and married and to have nonpsychotic disorders other than substance use disorders. The strongest and most consistent predictors of satisfaction were older age and better self-reported health. Longer length of stay was also associated with greater satisfaction on a majority of subscales. Findings among female and minority veterans were mixed across measures. Large facilities and facilities that specialize in mental health treatment had lower levels of satisfaction than others. Patient characteristics accounted for more of the variance in satisfaction than did facility characteristics. CONCLUSIONS: Older and healthier patients reported greater satisfaction with mental health care services. Accurate comparison of patient satisfaction between facilities requires that adjustments be made for differences in patient characteristics. Large facilities may need to make special efforts to personalize their services. PMID- 9406264 TI - Disruptive behavior and the determinants of costs in the public mental health system. AB - Efforts to increase the cost-effectiveness of public mental health systems are hindered by inadequate information about the determinants of use and cost. This paper reviews empirical research and theory suggesting that costs in the public health system are affected more by the disruptive behavior of persons with severe mental illness than by their age, sex, race, and diagnosis, which have been the focus of most economic studies. The author proposes modifications of traditional theories of health service use to explicitly account for the role of disruptive behavior in determining public mental health system costs. He describes a help seeking pathway in the public mental health system in which the decision to seek treatment is initiated not by the mentally ill person but by others affected by the person's disruptive behavior. This "other-determined" pathway into treatment is contrasted with the self-determined pathway in which an individual with distressing symptoms makes a rational choice to seek help. Empirical research consistent with the other-determined perspective will help target clinical interventions and system reforms to the factors responsible for high-cost mental health care and will improve the ability to predict resource use from observable clinical characteristics of consumers. PMID- 9406265 TI - Ethical benefits and costs of coercion in short-term inpatient psychiatric care. AB - OBJECTIVE: The study examined the outcome of psychiatric inpatient care in terms of patients' reports of ethical benefits, which were defined as fulfillment of the ethical principles of beneficence and autonomy, and ethical costs, which were defined as any violation of those principles. METHODS: A consecutive sample of 84 committed patients and a random sample of 84 voluntarily admitted patients in psychiatric care in two Swedish counties were studied. The patients were assessed twice by a psychiatrist, at admission and at discharge or after three weeks of care. They were also interviewed by a clinical psychologist at discharge or after three weeks. Four aspects of the ethical benefits or costs of their care were examined--whether they reported improvement in mental health, being treated with respect, not being violated as a person, and not being exposed to measures against their will (aside from commitment). RESULTS: The great majority of all patients reported improvement as a result of the psychiatric care. A third of the committed patients and more than half of the voluntarily admitted patients experienced ethical benefits only, without ethical costs. Twenty-three percent of the committed patients and 13 percent of the voluntary patients experienced ethical costs only, without ethical benefits. Some of the patients who experienced ethical costs only were also rated by a psychiatrist as not improved. CONCLUSIONS: Few patients had no measurable benefits of care. For committed as well as voluntary patients, an association was found between perceived respect for autonomy and self-reported improvement in mental health. PMID- 9406266 TI - Standard olanzapine versus placebo and ineffective-dose olanzapine in the maintenance treatment of schizophrenia. AB - OBJECTIVE: Two studies compared the efficacy of standard-dose oral olanzapine (5 to 15 mg a day) with placebo and with ineffective-dose olanzapine (1 mg a day) in maintenance therapy of schizophrenia. METHODS: The studies were 46-week double blind extensions of multicenter studies that assessed the efficacy of olanzapine in the acute treatment of schizophrenia. Subjects were 120 adults who met DSM-III R criteria for schizophrenia with an acute exacerbation and who had a minimum score of 24 on the Brief Psychiatric Rating Scale, who had responded to acute therapy (defined as at least a 40 percent reduction in the BPRS score from baseline or a score of 18 or less during up to six weeks of treatment), and who were outpatients at their last acute-phase visit. Relapse was defined as hospitalization for psychopathology. Relapse risk was analyzed using Kaplan-Meier survival analysis and life table analysis. Patients who relapsed were discontinued from the studies. RESULTS: In the first study (N = 58), patients in the standard-dose olanzapine group experienced a significantly lower relapse risk (p = .002) over one year than patients treated with placebo. The estimated one year risk of relapse with olanzapine was 28.6 percent, compared with 69.9 percent with placebo. Results were similar in the second study (N = 62); patients treated with standard-dose olanzapine had a significantly reduced risk of relapse (p = .018) over one year compared with patients treated with ineffective-dose olanzapine. The estimated one-year risks of relapse were 19.6 percent for standard-dose olanzapine and 45.5 percent for ineffective-dose olanzapine. CONCLUSIONS: Olanzapine is superior to placebo and ineffective-dose olanzapine in the maintenance therapy of schizophrenia. PMID- 9406267 TI - A spreadsheet method for calculating maximum caseload and intake capacity for CMHC psychiatrists. AB - OBJECTIVE: A method was sought to help administrators of community mental health centers determine a level of psychiatric staffing that is both cost-efficient and ensures high quality of care. METHODS: A survey of staff psychiatrists was conducted at a large community mental health center with seven outpatient clinics. The survey measured variables that can affect staffing requirements, including the number of hours psychiatrists have available for direct care, their preferred intervals between a patient's return visits, and the duration of appointments for an initial psychiatric assessment and for medication maintenance. A computer spreadsheet was developed to calculate the caseload capacity and intake capacity for clinics of the center. RESULTS: The survey indicated that the psychiatrists at the center had an average of 33 hours a week available for direct care. The mean preferred time between a patient's medication maintenance visits was 7.3 weeks. The mean time required for a psychiatric assessment was 80 minutes, and for a medication maintenance visit it was 33 minutes. With these data, the spreadsheet method was used to calculate intake and caseload capacity for psychiatric staff at three of the center's clinics. CONCLUSIONS: The data-based approach to calculating capacity can be modified to meet local needs. It brings objectivity to decision making about staffing, and the methods can improve resource management and enhance relationships between stakeholders and physicians. PMID- 9406268 TI - A 30-year progress report on a VA satellite psychiatric clinic program. AB - This paper describes a program of rural satellite psychiatric clinics that has evolved within a Veterans Affairs medical center over the past three decades. The eight clinics are staffed by a single team that includes a psychiatrist, a psychiatric nurse, and two mental health technicians. The team travels to the eight sites at least once each month. The program has achieved its goal of providing outpatient screening, diagnosis, and treatment services to veterans who would otherwise have no access to specialty psychiatric care due to distance and transportation problems. PMID- 9406269 TI - Instability before discharge and previous psychiatric admissions as predictors of early readmission. AB - In a retrospective chart review, the quality of care given 531 hospitalized veterans during an index admission was examined to determine whether patient characteristics or aspects of treatment differed between 243 patients readmitted within 30 days of discharge and a control group of 288 patients not readmitted for at least six months. The strongest predictor of early readmission was a greater number of previous admissions. Patient instability in the five days before discharge also predicted early readmission. These results may help identify patients at higher risk of early rehospitalization. Ensuring that patients with higher rates of rehospitalization are stable before discharge may help reduce early readmission. PMID- 9406270 TI - Care of long-term mentally ill patients by British general practitioners. AB - In the United Kingdom, patients gain access to psychiatric care through general practitioners (GPs). The first of three studies conducted to assess the role of GPs in managing patients with long-term mental illness found that such patients were unevenly distributed in general practices and that GPs preferred to care for them in collaboration with psychiatric specialists. A more detailed study of 16 general practices yielded information on characteristics and care of long-term mentally ill patients, including a high rate of GP consultations for them. A third, controlled study examined the impact of teaching GPs to provide a structured assessment of long-term mentally ill patients every six months; after the intervention, only a small number of patients actually received such assessments. PMID- 9406271 TI - The impact of welfare reform as perceived by users of mental health services in New York City. AB - A total of 118 psychiatric outpatients, 43 percent of whom were foreign born, completed a 12-item questionnaire about the impact of the new federal welfare legislation. A majority of respondents were worried about the new law and believed that it would worsen their mental symptoms, their well-being, and the quality of life in their neighborhood. Nearly half felt that the law had already affected their mental symptoms. Foreign-born patients were significantly more worried about the law than U.S.-born patients. The results suggest that organized psychiatry and individual psychiatrists should become more involved in activities to diminish the impact of the welfare legislation on patients and their families. PMID- 9406272 TI - A study of religion and psychotherapy. PMID- 9406273 TI - HIV risk behaviors. PMID- 9406275 TI - Scientology and APA. PMID- 9406276 TI - Readiness for rehabilitation. PMID- 9406277 TI - Need for integrated medical and psychiatric care emphasized in consensus statement on Alzheimer's. PMID- 9406278 TI - Operating principles for managed care, substance abuse approved by APA. PMID- 9406279 TI - Medication algorithms for use with public-sector patients being developed, tested in Texas project. PMID- 9406280 TI - Impairment of hepatic transport processes in perfused rat liver by the specific CCK receptor antagonist loxiglumide. AB - The specific cholecystokinin (CCK) receptor antagonist loxiglumide has been used in several human and animal studies to investigate the role of CCK in gastrointestinal physiology. In the present study, the interference of this CCK receptor antagonist with hepatic transport processes was characterized in the perfused rat liver. Indocyanine green, an organic dye which is secreted into bile without being metabolized, was taken up in control experiments at a rate of 68.1 +/- 7.7%. The CCK receptor antagonist lowered the extraction to 0.5 +/- 2.6% (P < 0.001). The compound diminished the hepatic extraction of CCK-8 from 90.95 +/- 2.60% to 4.90 +/- 1.95% (P < 0.001) and of gastrin from 22.2 +/- 1.1% to 8.2 +/- 1.9% (P < 0.001). The hepatic extraction of lidocaine, which is metabolized by the cytochrome P450 system, was only slightly altered. For leukotrienes and taurocholate, the rate-limiting step for transport into bile is secretion across the canalicular membrane; the hepatic extraction of leukotriene D4 was markedly diminished by loxiglumide whereas the transport of taurocholate was only slightly inhibited. The present study demonstrates that the specific CCK receptor antagonist loxiglumide diminished the hepatic extraction of various substances, including peptides and organic anions. It did not interfere with the cytochrome P450 system. The pronounced reduction of hepatic uptake of indocyanine green and leukotriene may be due to an interference with the transport system of these substances in the liver. PMID- 9406281 TI - Nitecapone is of benefit to functional performance in experimental heart transplantation. AB - In heart transplantation, global ischemia of a graft is followed by reperfusion injury. The formation of oxygen free radicals induces arrhythmias and impairs functional recovery of the graft. This study was executed to evaluate the effect of the new antioxidant, nitecapone, on ischemia-reperfusion injury in heart transplantation in rats. Donor hearts were perfused and stored at +4 degrees C for 2 h in either Ringer's solution in the control group (C-group, n = 26) or Ringer's solution with nitecapone (NC) added (NC-group, n = 18). The donor aorta was anastomosed to the recipient's abdominal aorta and the pulmonary artery to the recipient's inferior vena cava. The grafts were classified into three categories based on the functional recovery. The rats in both groups were killed at 10, 30, or 60 min after release of the aortic clamp. Tissue samples for chemiluminescence were obtained from the left ventricle, the right ventricle, and the septum of the heart. All grafts in the NC-group (18/18) began beating after release of the aortic clamp, whereas only 50% (13/26) of the grafts in the C group recovered (P < 0.0004). Chemiluminescence analysis showed lipid peroxidation values to be higher in the C-group than the NC-group up to 1 h after reperfusion. Also, the right ventricle samples showed lower chemiluminescence values in the NC-group than in the C-group. In conclusion, our results do not support the theory that different regions of the heart have different vulnerability to ischemia-reperfusion injuries. Nitecapone has a beneficial effect on the preservation of the grafts in terms of functional recovery. PMID- 9406282 TI - The effect of selective sympathetic denervation on pancreatic exocrine secretion. AB - The purpose of this study was to investigate the influence of selective sympathetic denervation of the rat pancreas on exocrine secretion and to study whether the observed effects were due to pancreatic trophism. Sprague-Dawley rats were divided into two groups. One group underwent selective sympathetic denervation by skeletonizing the superior and inferior pancreaticoduodenal and splenic arteries. The other group underwent simple laparotomy and served as controls. One week after the operation a catheter was introduced into the bile pancreatic duct and pancreatic juice was collected at 30-min intervals for 4 h. The output of bicarbonate, total protein, amylase, trypsin, chymotrypsin, lipase, colipase and carboxyesterlipase were determined. Following denervation secretion of pancreatic enzymes was significantly enhanced compared with sham-operated animals. We did not find any signs of pancreatic trophism 1 week after denervation. PMID- 9406283 TI - Evaluation of 99mTc-citrate, 67Ga-citrate and 99mTc(V) dimercaptosuccinic acid for the scintigraphic visualization of acute appendicitis. AB - An experimental study was planned to evaluate 99mTc-citrate, 67Ga-citrate and 99mTc(V) dimercaptosuccinic acid (DMSA) as agents for the visualization of acute appendicitis. Appendiceal ligation was performed through a midline incision in 24 rabbits. Twenty-four hours later the animals were divided into three equal groups. The rabbits were injected through the aurical vein with 1 mCi (37 MBq) 99mTc-citrate in group I, 0.5 mCi (18.5 MBq) 67Ga-citrate in group II and 1 mCi (37 MBq) 99mTc(V) DMSA in group III. After 3 h, static images of the rabbits were obtained with a gamma camera. There were positive images in seven, six and five rabbits in groups I, II and III respectively. The image quality was better in group I than in the other groups. Also, the mean uptake in group I was significantly higher than those of other two groups (P < 0.05). There was no significant difference between groups II and III (P > 0.05). All rabbits had appendicitis confirmed histologically. In conclusion, these results show that 99mTc-citrate is preferable to 67Ga-citrate and 99mTc(V) DMSA for the differential diagnosis of acute abdominal inflammations such as appendicitis, because of higher concentration ratios, simple and rapid preparation, low cost, excretion mainly through the kidneys and fast blood clearance. PMID- 9406284 TI - Regulation of mouse immuno-responses by a natural suppressor cell clone from bone marrow. AB - Natural suppressor (NS) activity is mediated by several kinds of cell lineages in bone marrow. We demonstrated that a NS cell, clone 7-31, was obtained after limiting dilution with bone marrow culture supplemented with WEHI supernatant. Clone 7-31, was capable of non-specific suppression of the generation of cytotoxic T lymphocytes without major histocompatibility complex restriction. Suppression of cytotoxic thymus-dependent lymphocyte (CTL) generation was also induced with a cell-free supernatant from the 7-31 cells. Interleukin-2 (IL-2) containing concanavallin-A-conditioned medium could not reverse the suppression. The supernatant did not inhibit the proliferation of IL-2-dependent CTLL-2 cells and rapidly growing tumour cells and fibroblasts. Thus, this bone marrow suppressor cell produces a soluble factor that inhibits the generation of CTL in vitro, and prolongs the acceptance of skin allografts in in vivo. PMID- 9406285 TI - Food as a source of polycyclic aromatic carcinogens. AB - Polycyclic aromatic hydrocarbons (PAH) belong to a large chemical family comprising many different compounds with important biological activity in mutagenic and carcinogenic processes. PAH have been detected in both raw and processed foods. The presence of PAH in non-processed foods is associated with environmental pollution from both human and industrial activities, whereas contamination of processed foods can be caused by certain preservation and processing procedures. Both toxicological and epidemiological studies have shown a relation between such compounds and tumor development. The data indicate that PAH must undergo a biotransformation process that causes the formation of biologically active metabolites. In this process, the presence of an enzyme complex that is induced by different xenobiotics is implied, making the toxicity of such compounds hard to predict. As setting a threshold limit below which toxicity could be considered negligible is difficult, the presence of PAH in foodstuffs should be reduced to as low as possible by controlling environmental contamination and all procedures that could cause PAH contamination during food processing, preserving, and packaging. PMID- 9406286 TI - Environmental uranium and human health. AB - Uranium from the environment enters the human body by ingestion with food and drink and by inhalation of respirable airborne uranium-containing dust particles or aerosols. Daily intake of uranium in food and water varies from approximately 1 to approximately 5 micrograms U/d daily in uncontaminated regions to 13-18 micrograms/d or more in uranium mining areas. A 70 kg, non-occupationally exposed 'Reference Man' living in Europe or in the United States has an estimated total body uranium content of about 22 micrograms. Uranium is absorbed from the intestine or the lungs, enters the bloodstream, and is rapidly deposited in the tissues, predominantly kidney and bone, or excreted in the urine. In the bloodstream, uranium is associated with red cells, and its clearance is relatively rapid. Renal toxicity is a major adverse effect of uranium, but the metal has toxic effects on the cardiovascular system, liver, muscle, and nervous system as well. Any possible direct risk of cancer or other chemical- or radiation-induced health detriments from uranium deposited in the human body is probably less than 0.005% in contrast to an expected indirect risk of 0.2% to 3% through inhaling the radioactive inert gas radon, which is produced by the decay of environmental uranium-238 in rocks and soil and is present in materials that are used to build dwellings and buildings where people live and work. PMID- 9406287 TI - Poverty, health, and nutrition in Germany. AB - OBJECTIVE: To investigate the relation between poverty and several variables describing health and nutrition behavior in East Germany and West Germany. METHODS: Data are from the third National Health Survey in West Germany and the first Health Survey for the new federal states of Germany (1991/92). Both health surveys included a self-administered questionnaire ascertaining sociodemographic variables, smoking history, nutritional behavior (using a food-frequency list), physical activity, and a medical examination comprising measurements of height, weight, blood pressure, and blood sampling for serum cholesterol determination. Participants included 4958 subjects in the West Survey and 2186 subjects in the East Survey aged 25-69 years, with a respective net response rate of 69.0% and 70.2%. Poverty was defined as a household equivalence income of 62.5% or less of the median income of the general population. RESULTS: The lowest income group (poverty or near poverty) comprised 11.6% of East German versus 15.9% of West German males and 14.8% of East German versus 19.3% of West German females. For most but not all health and nutrition parameters, less favorable results were obtained for subjects with an equivalence income below or near poverty. The most striking poverty-related differences regarding cardiovascular disease risk factors were found for lack of regular exercise for both genders and obesity in females. No poverty-related differences were found for the prevalence of hypercholesterolemia, despite a much higher prevalence of obesity in persons with an income below the poverty line. Current nutritional behavior and changes in nutritional behavior during the last three years was strongly related to income status, with a more unhealthy status for low-income population groups in both East and West Germany. CONCLUSIONS: In Germany, poverty has strong effects on individual health status and nutritional behavior. Because of rising unemployment rates and reductions in social security payments for low-income groups, it is likely that the negative consequences of poverty on health are increasing. PMID- 9406288 TI - Effect of 50-Hz electromagnetic field on the retention of toxic radionuclides in rat tissues. AB - The effect of electromagnetic field (EMF) 50 Hz, 10 mT, on the tissue retention of radiotoxic polonium-210 and thorium-234 was studied in a rat model. Regarding 210Po in the ionic state, small but significant effects were obtained by exposure of rats to EMF either before the intravenous injection of 210Po (pre-exposure) or after the rats had already been injected with 210Po (post-exposure). When compared with control values, pre-exposure to EMF caused a significant 28% decrease in the retention of 210Po in the skin and a 10% decrease in total 210Po retention in the investigated tissues. Relative to controls, post-exposure resulted in a 131% increase in 210Po retention only in the thymus. Regarding carrier-free 234Th in the ionic state, both types of EMF exposure caused a substantial increase in 234Th retention in the liver and spleen and a decrease of 234Th in the bones. A different effect of EMF on the retention of 234Th in the body was obtained when the mass of thorium was increased by adding as carrier 232Th (50 micrograms kg-1 body mass). With pre-exposure, a significant 10% decrease in the high retention of 234Th in the liver (77% of injected radioactivity) was observed. On the other hand, with post-exposure no significant changes in retention of 234Th were found in the tissues. PMID- 9406289 TI - Molecular epidemiology: cancer risk assessment using biomarkers for detecting early health effects in individuals exposed to occupational and environmental carcinogens. AB - Detecting changes that precede the overt symptoms of cancer and identifying measurable indices of such changes in persons exposed to occupational and environmental carcinogens constitutes one of the primary objectives of molecular epidemiology research. Biomarkers represent a valuable research tool that makes it possible to attain that objective. Suitably selected biomarker sets may provide information on the extent of exposure to carcinogenic agents (internal dose, biologically effective dose), detect early changes caused by those agents in the exposed organism, and identify individuals with a particularly high risk of cancer development. The tremendous progress in research on the mechanisms of cancer initiation and promotion has enabled the assessment of cancer risk in healthy individuals by examining specific results from determinations of suitably selected biomarkers. The finding that gene defects (gene mutations and changes of their expression) constitute the background of carcinogenesis has resulted in molecular biology becoming focused on detecting defective genes or proteins synthesized under control of the defective genes. PMID- 9406290 TI - Public health impacts of global climate change. AB - The potential health impacts of climate change are wide-ranging, from direct impacts at familiar local scales, through indirect effects occurring at the regional or ecosystem level, to long term effects on the sustainability of global systems. To assess these potential impacts, there is a need to broaden the scope of health impact assessment. Eco-epidemiology is emerging as a response to this need. Eco-epidemiology entails a shift in focus: from direct (toxicological) to indirect (ecological) mechanisms; and from effects occurring at 'human' temporal and geographical scales to those at regional and geophysical scales. We discuss the potential health impacts of climate change on each scale. At the global scale, interactions and feedbacks between systems are critical determinants of long term outcomes. From an eco-epidemiological perspective, the study of climate change becomes inseparable from the study of global change more generally. PMID- 9406291 TI - Alcohol dependence: a critical look at the effects of alcohol metabolism. AB - This paper explores the metabolic consequences of alcohol misuse and identifies the pathophysiological reasons why alcohol, no matter what quantity is taken regularly, is not beneficial for the normal functioning of most body systems. Although moderate ethanol consumption may reduce stress and the risk of coronary heart disease, ethanol also exerts a direct toxicological effect because it interferes with hepatic metabolism and immune functions. Liver transplantation may be necessary for end-stage liver disease in alcoholics. A causal effect between alcohol intake and several cancers has been reported. Both environmental and genetic factors are involved in the susceptibility to alcoholism. An explanation of alcohol dependence as a family disease is introduced to shed light on the magnitude of its collateral effects on the family and on the community as a whole. The adverse effects of alcohol on pregnant women and the fetus are also discussed. To provide awareness of the effectiveness of community efforts, we examined the possible intervention strategies and the role of community care in this regard. PMID- 9406292 TI - Magnesium substitution and postoperative arrhythmias in patients undergoing coronary artery bypass grafting. AB - Sixty coronary artery bypass grafting patients were randomized to receive either magnesium sulphate or placebo for 4 days postoperatively. The magnesium substitution reduced the duration of atrial fibrillation or flutter (p < 0.05), but not the number of patients developing these arrhythmias. The number of ventricular ectopic beats was also reduced among patients receiving magnesium sulphate compared to placebo (p < 0.05). To evaluate whether the anti-arrhythmic effect of magnesium sulphate was explained by a faster resumption of cellular potassium postoperatively, skeletal muscle electrolyte concentrations were measured pre-operatively and on the third day postoperatively. No significant difference was found in skeletal muscle potassium or magnesium contents on the third day postoperatively when comparing the two groups. The serum magnesium level declined postoperatively in the placebo group, whereas an increase was found in patients receiving magnesium sulphate. We suggest magnesium substitution as a routine postoperatively, because this treatment seems to reduce the severity of postoperative arrhythmias. PMID- 9406293 TI - Influence of complete revascularization on long-term survival after coronary artery bypass surgery. AB - To evaluate the influence of complete coronary revascularization after coronary artery bypass grafting (CABG) on long-term survival, we reviewed the records, including reports of coronary angiography, of 198 patients (25 women). Coronary artery bypass grafting was performed in the period 1973-1982, when the patients' mean age was 52.5 years. No significant (p < 0.05) difference in survival in the first 15 postoperative years was found between the patients judged to have complete revascularization at coronary angiography 6 months after CABG and the general Danish population. Complete revascularization by extensive grafting should improve survival of patients treated for angina. If the revascularization remains complete at coronary angiography assessment 6 months postoperatively, the 15-year survival rate can be expected to equal that in the general population. PMID- 9406294 TI - Haemostasis at low heparin dosage during cardiopulmonary bypass with heparin coated circuits in pigs. AB - Cardiopulmonary bypass (CPB) causes activation of cascade systems. Although heparin coating of CPB circuits improves biocompatibility, the effects on coagulation remain controversial. Theoretically, heparin coating should permit the reduction of systemic anticoagulation during CPB. We investigated influences on haemostatic variables in animal CPB, comparing heparin-coated circuits and reduced systemic heparinization (group HC) with uncoated circuits and full heparinization (group C). Twenty pigs underwent 2-h CPB. Seven (HC, n = 4; C, n = 3) were weaned from CPB and studied for up to 4 h. Total administered heparin was 470 +/- 6 IU/kg (mean +/- SEM) in group C and 100 +/- 0 IU/kg in group HC. Protamine dosage was significantly reduced in group HC. In group C, levels of prothrombin complex, factor VIII and adhesive platelets were reduced significantly during CPB, and postoperatively there were significantly lower values of prothrombin complex, fibrinogen, antithrombin III, factor VIII and adhesive platelets but a significantly increased concentration of von Willebrand factor and cumulative bleeding after 4 h. In conclusion, heparin-coated CPB circuits combined with lowered heparin dosage reduced coagulation factor consumption and preserved platelet function, possibly contributing to improved postoperative haemostasis. PMID- 9406295 TI - Effect of methylprednisolone on the oxidative burst activity, adhesion molecules and clinical outcome following open heart surgery. AB - Following cardiac surgery with cardiopulmonary bypass (CPB), activated granulocytes may be involved with ischaemia/ reperfusion injury. The purpose of this study was to investigate whether steroids could reduce the oxidative burst activity of granulocytes, the expression of adhesion molecules on granulocytes and improve clinical outcome. Sixteen patients undergoing open heart surgery participated in the study. Eight were randomized to receive methylprednisolone (30 mg/kg intravenously) at the start of anaesthesia while eight patients served as a control group. The oxidative burst was measured flow cytometrically using 123-dihydrorhodamine. A panel of adhesion molecules was measured using monoclonal antibodies. Following CPB the oxidative burst activity and the expression of the adhesion molecule L-selectin more than doubled compared to initial values. There was no difference between the steroid group and the control group regarding the expression of adhesion molecules or the oxidative burst activity. In the steroid group the fluid gain during extracorporeal circulation (ECC) was 683 ml (median) compared to 1488 ml in the control group. Steroids prevented hyperthermia in the postoperative period but did not improve the weaning from the ventilator or reduce the stay in the intensive-care unit. In conclusion, treatment with steroids prevented hyperthermia following open heart surgery with CPB and reduced capillary leak during ECC. Methylprednisolone, however, did not reduce the oxidative burst activity or the expression of adhesion molecules on granulocytes following CPB. PMID- 9406296 TI - Combined antegrade-retrograde blood cardioplegia does not protect right ventricle better than either technique alone in patients with occluded right coronary artery. AB - To study the hypothesis that combined antegrade-retrograde delivery of cardioplegia might overcome the limitations in myocardial protection of either technique alone, we compared the distribution of the different cardioplegic approaches by assessing myocardial cooling and evaluated the effects on right ventricular (RV) function in elective coronary artery bypass grafting (CABG) patients with occluded right coronary artery (RCA). In a randomized trial, 15 patients received exclusively antegrade (ante group), 14 patients received exclusively retrograde (retro group) and 15 patients received combined, alternating antegrade-retrograde (combi group) cold blood cardioplegia. Myocardial temperatures were measured at four sites in the heart. Right ventricular function was assessed by determining the ejection fraction (fast response thermodilution) and preload-related RV stroke work in repeated measurements. Myocardial cooling was similarly uneven and the posterior wall of the RV remained above 20 degrees C after all three methods of delivering hypothermic (5-7 degrees C) cardioplegia. The RV ejection fraction and preload related (right atrial pressure) RV stroke work decreased postoperatively similarly in all groups. The results suggest that combined antegrade-retrograde cold blood cardioplegia could not provide more homogeneous myocardial cooling or better RV recovery than either technique alone in three-vessel-diseased CABG patients with occluded RCA. PMID- 9406297 TI - Chronic total occlusions of coronary arteries--medical versus surgical treatment. AB - Coronary artery bypass grafting (CABG) is an established treatment of patients with angina pectoris and chronic total coronary occlusion of major coronary arteries. However, in patients with mild or absent angina and chronic total coronary occlusion, optimal treatment is unsettled. We compared the prognosis of patients with chronic total coronary occlusion treated medically because of mild or absent angina with a matched group of patients undergoing CABG. In a retrospective design we evaluated all coronary angiographies performed in our department over a 5-year period. We identified 77 patients with chronic total occlusion of major coronary arteries eligible for CABG but treated medically because of mild or absent angina. The medically treated patients were matched on age, sex and ejection fraction with 77 patients with occluded major coronary arteries and angina pectoris who were treated surgically. The main outcome measures were death, acute myocardial infarction (AMI) and CABG. At baseline, CABG patients demonstrated an increased duration and severity of angina pectoris and an increased consumption of anti-anginal drugs. No differences were found with regard to angiographic parameters. The 5-year event rates (medically treated versus CABG) were: death, 14% vs 7% (p = 0.08); death or AMI; 27% vs 16% (p = 0.10); death, AMI or CABG, 34% vs 16% (p = 0.03) (log-rank statistics). In conclusion, our data indicate that patients with chronic total coronary occlusions and mild or absent anginal symptoms may benefit from surgical treatment. PMID- 9406298 TI - A study of 47 bacteremic Staphylococcus aureus endocarditis cases: 23 with native valves treated surgically and 24 with prosthetic valves. AB - A retrospective review of medical records from the Staphylococcus Laboratory, Copenhagen, 1982-1991, was carried out at the Department of Clinical Microbiology, Statens Serum Institut, 1994-1995, to investigate the clinical features and outcome of two subgroups of bacteremic Staphylococcus aureus endocarditis cases in non-drug addicts: patients with prosthetic valve endocarditis (PVE) and patients with native valve endocarditis treated surgically. Twenty-four cases of PVE were included. Six cases were early (within 60 days of valve implantation) and 18 were late. The overall in-hospital mortality was 42%. Surgical treatment resulted in a non-significantly lower mortality as compared with medical treatment alone (0% vs 50%, p = 0.19). Medical treatment of aortic and mitral valve endocarditis resulted in similar mortality rates (44% and 50%, respectively). Twenty-three cases of native valve infective endocarditis had the valve replaced surgically. The in-hospital mortality was 22%, which was significantly lower as compared with medical therapy (69%, p < 0.0001). The treatment changed significantly during the study period: 6 of 112 patients (5%) were treated surgically in the first half of the period (1982-1986) compared to 17 of 124 patients (14%) in the second half (1987-1991, p = 0.049). Severe congestive heart failure was the main indication for cardiac surgery in 21 patients. In conclusion, a shift towards a more aggressive surgical approach has taken place in the 10-year period. This development should be strengthened in the future as surgical intervention may improve survival in patients with infective endocarditis caused by S. aureus whether the infected valve is prosthetic or native. PMID- 9406299 TI - Mediastinitis due to Mycobacterium fortuitum infection following Fontan operation in a child. AB - Here we report a case of mediastinitis due to Mycobacterium fortuitum infection in a child after a Fontan operation. To our knowledge this is the first report of atypical mycobacterial mediastinal infection after congenital heart surgery. Atypical mycobacteria can be the cause of "culture negative" sternal and thoracotomy wound infections. A brief review of the literature is included in the discussion. PMID- 9406300 TI - Reflux, stricture and glomerular filtration rate after two antireflux techniques in continent urinary reconstruction using the right colon. AB - In the present study the incidence of reflux, stricture formation and changes in glomerular filtration rate in patients with the submucosal tunnel or the Camey-Le Duc technique of ureteric implantation into the caecum/detubularized right colon used for continent cutaneous diversion/orthotopic bladder substitution was investigated. Reflux was found in two renal units and ureterointestinal stenosis occurred in five renal units after submucosal tunnel ureteric implantation. After Camey-Le Duc ureteric implantation, one renal unit showed reflux and none had stenosis of the ureterointestinal anastomosis during follow-up. Mean glomerular filtration rate (ml/min/1.73 m2) fell from 98 to 85 in the submucosal tunnel group and from 88 to 81 in the Camey-Le Duc group after mean follow-ups of 9 and 5 years, respectively. Both methods of ureteric implantation in this study were effective in preventing reflux, and renal function was well preserved in both groups. The absence of ureterointestinal strictures in the Camey-Le Duc group is encouraging and indicates that this is a reliable method for reflux prevention. PMID- 9406301 TI - One or two incisions for nephroureterectomy in transitional cell renal pelvis tumours. AB - A retrospective study comprising 18 patients with transitional cell renal pelvis tumours (TCPT) was carried out to evaluate the results after two different surgical procedures for nephroureterectomy. The kidney was removed by a flank incision and the lower part of the ureter by either an incision in the lower part of the abdomen or intussusception of the ureter followed by transurethral resection of the ureteral orifice. Eight patients were subjected to nephroureterectomy by means of two incisions and another eight patients underwent a simple nephrectomy followed by ureteral intus-susception and transurethral resection. Two patients received other treatments. After nephroureterectomy with a separate incision for ureterectomy, the average hospital stay was 12 days, compared with 7.5 days in patients operated upon with only one abdominal incision. Recurrence of tumour or survival was not significantly different in the two groups. PMID- 9406302 TI - Local excision of ureteral tumours. AB - A retrospective study was carried out on 20 patients with transitional cell ureteral tumours (TCUT). Surgical exploration of the tumour was performed in 17 patients. Biopsies for frozen section were evaluated for decision concerning the extent of operative intervention. If radical excision of the tumour could be done, and if the ureteral defect could be bridged, a conservative procedure was chosen. Thus, segmental resection of the ureter and primary end-to-end closure of the ureter was performed in 5 patients and ureteroneocystostomy in 3 patients. In nine patients local tumour excision was not feasible and nephroureterectomy was done. The survival rate at 3 and 10 years after ureteral resection as well as after nephroureterectomy was the same, 66% and 16%, respectively. Local excision of non-invasive low-grade ureteral tumours could be safely performed in selected patients, based on local findings and frozen section at the time of surgery. PMID- 9406303 TI - Structure of uric acid concretion developed around a catheter. AB - A uric acid concretion formed round a catheter (JJ stent) in the bladder and removed intact from the body together with the catheter was studied using an electron scanning microscope. The concretion was composed of anhydrous uric acid, some uric acid dihydrate (< 5 wt.%) and individual particles of calcium oxalate monohydrate. The stone interior was porous with frequent occurrence of differently sized cavities that were either empty or partially filled with particles of uric acid and/or calcium oxalate monohydrate. Calcium oxalate particles were not of crystalluria origin but developed in the cavity. The succession of processes leading to the stone formation was deduced from its inner structure. The stone was formed due to a crystalline growth with minor, if any, participation of sedimentation. The estimated average rate of the calculus development, 2 x 10(-9) m/s, confirms the predominant role of crystalline growth in stone formation and indicates a relatively low urinary supersaturation with respect to uric acid prevailing during the period of calculogenesis. PMID- 9406304 TI - Rhenium-186 HEDP: palliative radionuclide therapy of painful bone metastases. Preliminary results. AB - The aim of this study was to evaluate the efficacy of rhenium-186 hydroxyethyledine diphosphonate (Re-186 HEDP) for pain relief in patients with disseminated bone metastases primarily from prostate or breast cancer. Up to now, 44 patients taking analgesics were entered in this study and received one or more injections of 1295 MBq of Re-186 HEDP. An analgesic effect of more than 20%, evaluated by using a verbal rating scale (VRS) and a visual analogous scale (VAS), was defined as significant and could be achieved in 60% of these patients. Duration of clinical response averaged 1-4 months (median 5.5 weeks). Side effects such as a moderate decrease of platelets or an increase of pain for 1-2 days (flare-up effect) were observed. Radioactive treatment with Re-186 HEDP appears to be a useful compound for the palliation of painful skeletal metastases and improvement of the remaining quality of life. PMID- 9406305 TI - Evaluation of non-invasive clinical samples in chronic chlamydial prostatitis by using in situ hybridization. AB - Seventy-eight non-invasive prostate specimens collected from patients with chronic non-bacterial prostatitis were evaluated by in situ hybridization (IH) for evidence of Chlamydia trachomatis. Intracellular Chlamydia bodies were detected in 18 of 78 cases (20.6%). Homogeneous blue-black bodies in the cellular cytoplasm were accepted as in situ positive. Chlamydial antigen detected by enzyme immuno assay (EIA) was positive in 12 cases (13.7%), but only nine of them were positive by IH. Our study confirms previous reports implicating C. trachomatis as an aetiological agent in chronic non-bacterial prostatitis, and underscores the applicability of DNA probes for detection and identification of C. trachomatis in prostatic materials. PMID- 9406306 TI - Lichen sclerosus et atrophicus. Findings after complete circumcision. AB - We prospectively evaluated 75 patients, 30-77 years old, with severe phimosis. All patients were treated surgically by complete circumcision and all surgical specimens were sent for histological evaluation. All patients with histologically proven lichen sclerosus et atrophicus (LSA) (eight patients, 10.6%) were re evaluated 6 months postoperatively, and all but one were examined 5 years after the operation. All patients with histologically proven balanoposthitis (BP) (47 patients, 62.6%) were also re-evaluated 6 months postoperatively, and 41 patients 5 years after surgery. This group (BP) was the control group of our study. Six months after the operation, the eight patients with histologically proven LSA all had an excellent convalescence, and the lesions observed during the operation resolved in four patients and regressed in two patients. In one patient the glans presented with a pale grey-white-coloured plaque. Biopsy was performed and a well differentiated squamous cell carcinoma infiltrating the glans was revealed. The control group of patients with histologically proven BP were also re-evaluated 6 months and 5 years after surgery. An excellent convalescence was observed in all patients who completed the follow-up examination. Care must be taken not to underestimate the potential relationship between LSA and squamous cell carcinoma, because the latter is usually invasive, very aggressive and requires immediate treatment. PMID- 9406307 TI - Penile tourniquet: an invaluable technique. AB - In 72 cases of minor penile surgeries, including circumcision (42), meatotomy (18), meatoplasty (8) and chordee correction (4), a ring cut from a surgical glove finger was used as a tourniquet at the base of the penis after infiltrating the skin and the glans with plain 1% xylocaine. The usual surgical procedures were performed and the tourniquet was released after securing the probable bleeding points. The postoperative results were excellent, with oedema of the penis being the only complication. There was no incidence of skin necrosis, sensory impairment, wound infection, erectile dysfunction or postoperative oozing. This bloodless technique not only curtailed the operating time and made the procedure simpler, but it also prevented the washout of xylocaine by impeding venous drainage. PMID- 9406308 TI - Short-arm dicentric Y chromosome associated with Sertoli-cell-only tubule. AB - We report a case of a short-arm dicentric Y chromosome associated with Sertoli cell-only tubule. Chromosomal analysis, using G- and C-banding techniques, revealed: 45,X/46,X psu dic(Y) (pter-->q11::q11-->pter)/46,X + mar. Staining by fluorescence in situ hybridization using a Y chromosome centromere-specific DNA probe showed two bright spots in the pseudodicentric Y chromosome and one in the marker chromosome. It is assumed that Sertoli-cell-only tubule is caused by deletion or disruption of the azoospermic factor gene located distal in Yq11. PMID- 9406310 TI - A prospective study of transperitoneal transport in patients undergoing peritoneal dialysis. AB - A prospective one-year follow-up study of the functional characteristics of the peritoneal membrane was conducted in 20 peritoneal dialysis patients. Ten patients had at least one episode of peritonitis during the follow-up period. Changes in the transperitoneal transport of small solutes were evaluated by the mass transfer area coefficients (urea, creatinine, and glucose), ultrafiltration sieving coefficients (urea and creatinine), and peritoneal equilibration test results. Changes in the capacity of the peritoneal membrane to transport macromolecules were evaluated by means of albumin mass transfer rates. Finally, changes in transperitoneal water transport were evaluated by means of the ultrafiltration properties and the lymphatic flow rates. After one year of follow up, transport of water and the investigated solutes had not changed significantly. Even episodes of peritonitis had no permanent influence on the transport function of the peritoneal membrane. The intraperitoneal residual volumes before instillation were significantly larger in patients who developed peritonitis during the follow-up period compared to patients who did not. Over a one-year period, no changes in peritoneal membrane characteristics could be demonstrated. A large residual volume of dialysate may be a marker of increased risk of getting peritonitis. PMID- 9406311 TI - The arginine-nitric oxide pathway in thrombotic microangiopathy. AB - Eleven patients with thrombotic thrombocytopenic purpura (TTP) or haemolytic uremic syndrome (HUS) were investigated with respect to plasma concentrations of L-arginine, a substrate for nitric oxide (NO) and asymmetrical dimethyl arginine (ADMA), during active disease and after recovery. Plasma concentration of NO3-, the degradation product of NO, was also analyzed. The patients were treated with fresh-frozen plasma and plasmapheresis. One of the patients had experienced relapses of TTP five times during the preceding year. After treatment with p.o. arginine hydrochloride 1.5 g x 3 was started, no relapse has occurred during a 12 month period. During the active phase the plasma concentration of arginine was low and that of NO3- was very high, indicating a high NO-synthesis rate. The arginine concentration normalized on recovery. Plasma levels of ADMA, was twice normal during active disease, and did not return to normal on recovery. In conclusion, patients with TTP/HUS exhibit signs of activation of the NO synthesis. PMID- 9406309 TI - Prevalence of autoantibodies associated with glomerulonephritis, unaffected after extracorporeal shock wave lithotripsy for renal calculi, in a three-year follow up. AB - Extracorporeal shock wave lithotripsy (ESWL) has become a useful tool in the treatment of renal calculi, but side effects may occur. Hitherto, two case reports have been published of an anti-glomerular basement membrane disease resulting in end-stage renal failure following ESWL treatment. In this prospective study of 59 consecutive patients undergoing ESWL for renal calculi, the prevalence of autoantibodies associated with glomerulonephritis was investigated before ESWL and at 3-year follow-up. The prevalences of antinuclear, anti-glomerular basement membrane, anti-neutrophil cytoplasmic and thyroid antibodies were found to be within the respective normal ranges prior to the first ESWL treatment and to be unaffected at follow-up. PMID- 9406312 TI - Allergic nephropathy associated with norfloxacin and ciprofloxacin therapy. Report of two cases and review of the literature. AB - Allergic nephropathy associated with quinolone antibiotics has been reported in an increasing number of cases. The mechanism might be a hypersensitivity reaction. Norfloxacin has been incriminated previously as a cause once only, with acute interstitial nephritis (AIN) as the histopathological finding. Ciprofloxacin-associated nephropathy has been reported in 28 cases, with AIN as the main histopathological finding. This report describes a second case of AIN associated with norfloxacin treatment and another ciprofloxacin-associated renal interstitial drug adverse reaction. Clinicians should be aware of quinolone associated AIN, which is a rare but potentially dangerous renal complication. PMID- 9406313 TI - The role of magnetic resonance spectroscopy in the assessment of kidney viability. AB - Renal transplant programmes are seriously limited by the continuing shortage of donor organs. Kidneys from marginal and non-heart-beating donors are increasingly being used, but their viability may be compromised. There is currently no rapid yet accurate method for assessing donor organ viability which can be applied within the window of opportunity between harvesting and implantation. Magnetic resonance spectroscopy (MRS) is a non-invasive technique which is being increasingly applied to delineate biochemical changes in vivo. Studies in animal models and humans now suggest that phosphorus-31 MRS may be useful in the non invasive assessment of isolated donor kidney viability. PMID- 9406314 TI - It's like a pain in the ... perineum: a surgical clip protruding into the urethra through the urethrovesical anastomosis after radical prostatectomy. AB - A 69-year-old male underwent a radical retropubic prostatectomy for prostate cancer. He presented severe perineal pain that prevented him from sitting normally after removal of the urethral catheter. Cystography and echography disclosed a possible aetiology that was later confirmed by cystoscopy: a metal clip protruding into the urethra through the urethrovesical anastomosis. PMID- 9406315 TI - Interstitial cystitis and bladder mastocytosis in a woman with chronic urticaria. AB - A patient with chronic urticaria and angioedema developed endoscopically confirmed interstitial cystitis. Bladder biopsy revealed bladder mastocytosis with > 60 mast cells/mm2 (normal < 10) in the detrusor and submucosa. Most of the mast cells were activated and degranulated. The occurrence of IC in a patient with urticaria and angioedema, diseases both associated with mast cell pathophysiology, supports the role of mast cells in interstitial cystitis. PMID- 9406316 TI - Paraganglioma of the prostate. A case report and review of the literature. AB - Only five cases of paraganglioma of the prostate have been reported hitherto. The tumours may be functional (phaeochromocytomas) or non-functional and benign or malignant. We present a sixth case--a non-functional, benign paraganglioma of the prostate--with a review of the literature. PMID- 9406318 TI - Pyelitis cystica. AB - We present four cases of pyelitis cystica. Aetiology, pathogenesis, diagnoses, treatment and differential diagnosis are discussed. We recommend follow-up until malignant disease has been excluded. PMID- 9406317 TI - Eighteen-month follow-up of cystovaginoplasty: a new augmentation operation for repair of vesicovaginal fistulae. AB - We present an 18-month follow-up of a cystovaginoplasty case. Surgery was performed because of a large vesicovaginal fistula. A new technique using the vagina as a pouch was utilized. PMID- 9406319 TI - Arousal and cardiorespiratory responses to airflow obstruction in sleeping lambs: effects of sleep state, age, and repeated obstruction. AB - We studied the effects of postnatal age on arousal and cardiorespiratory responses to airflow obstruction in sleeping lambs: we also determined the influence of sleep states and repeated airflow obstruction. Sixteen lambs were chronically prepared for monitoring sleep states, arterial O2 saturation (SaO2), heart rate (HR), and intrapleural pressure (Pp1) and were studied from 2-29 days after birth. Obstruction of respiratory airflow by facemask occlusion led to arterial desaturation, augmentation of respiratory efforts, bradycardia, and arousal. Lambs aroused more rapidly and with less desaturation in non-rapid eye movement (NREM) sleep (7 +/- 1 second and 7 +/- 1%, respectively) than in rapid eye movement (REM) sleep (18 +/- 2 seconds and 22 +/- 2%), and cardiac slowing was less in NREM than in REM sleep. In REM sleep only, the arousal latency and desaturation at arousal were affected by postnatal age; arousal responses occurred most rapidly in the youngest (< or = 6 days) and oldest (> or = 13 days) age groups and were delayed at 7-12 days. Repeated episodes of airflow obstruction led to reduced arousability in REM sleep only. We conclude that arousal from REM. but not NREM, sleep in response to the obstruction of respiratory airflow is transiently depressed during early postnatal development and that repeated obstructions and arousals also lead to depressed arousal from REM sleep. PMID- 9406320 TI - Abnormal visual P300 latency in obstructive sleep apnea does not change acutely upon treatment with CPAP. AB - This study evaluated the effects of 2-4 months of continuous positive airway pressure (CPAP) treatment on previously demonstrated P300 latency prolongations in obstructive sleep apnea (OSA). Subjects with severe OSA (respiratory disturbance index > 40/hour sleep) were administered polysomnograms, auditory and visual P300 testing using 31 scalp electrodes, and multiple sleep latency testing before and after treatment with CPAP for 2-4 months. Despite significant improvements in sleep and respiratory variables and the mean sleep latency, there were no significant P300 changes. Obstructive sleep apnea patients had prolonged visual P300 latency compared to normals, both before and after treatment. Prolongations in P300 latency that are resistant to the acute effects of CPAP may suggest that OSA causes physiological cortical changes that are unrelated to sleepiness and may be resistant to treatment. PMID- 9406321 TI - Estimation of the clinically diagnosed proportion of sleep apnea syndrome in middle-aged men and women. AB - The proportion of sleep apnea syndrome (SAS) in the general adult population that goes undiagnosed was estimated from a sample of 4,925 employed adults. Questionnaire data on doctor-diagnosed sleep apnea were followed up to ascertain the prevalence of diagnosed sleep apnea. In-laboratory polysomnography on a subset of 1,090 participants was used to estimate screen-detected sleep apnea. In this population, without obvious barriers to health care for sleep disorders, we estimate that 93% of women and 82% of men with moderate to severe SAS have not been clinically diagnosed. These findings provide a baseline for assessing health care resource needs for sleep apnea. PMID- 9406322 TI - Ventilatory response to induced auditory arousals during NREM sleep. AB - Sleep state instability is a potential mechanism of central apnea/hypopnea during non-rapid eye movement (NREM) sleep. To investigate this postulate, we induced brief arousals by delivering transient (0.5 second) auditory stimuli during stable NREM sleep in eight normal subjects. Arousal was determined according to American Sleep Disorders Association (ASDA) criteria. A total of 96 trials were conducted; 59 resulted in cortical arousal and 37 did not result in arousal. In trials associated with arousal, minute ventilation (VE) increased from 5.1 +/- 1.24 minutes to 7.5 +/- 2.24 minutes on the first posttone breath (p = 0.001). However, no subsequent hypopnea or apnea occurred as VE decreased gradually to 4.8 +/- 1.5 l/minute (p > 0.05) on the fifth posttone breath. Trials without arousal did not result in hyperpnea on the first breath nor subsequent hypopnea. We conclude that 1) auditory stimulation resulted in transient hyperpnea only if associated with cortical arousal; 2) hypopnea or apnea did not occur following arousal-induced hyperpnea in normal subjects; 3) interaction with fluctuating chemical stimuli or upper airway resistance may be required for arousals to cause sleep-disordered breathing. PMID- 9406323 TI - How a general population perceives its sleep and how this relates to the complaint of insomnia. AB - The traditional indicators of insomnia (i.e. difficulty initiating sleep, difficulty maintaining sleep, nonrestorative sleep, early morning awakening) were assessed in a representative sample of 1,722 French-speaking Montrealers (Canada) aged 15 to 100 years. These subjects were interviewed over the telephone (81.3% of contacted sample) by means of the Sleep-Eval software. Subjects were classified as either satisfied or dissatisfied with quality of sleep (SQS or DQS), with or without insomnia indicators (+I or -I). Sociodemographics, sleep wake schedules, evening activities, medication intake, recent medical consultations, and social life were also investigated. DQS subjects composed 17.8% of the population (DQS + I: 11.2%; DQS - I: 6.5%), and 21.7% of subjects were classified as either DQS + I or SQS + I. Overall, 3.8% of subjects reported using a sleep-enhancing medication. Nonrestorative sleep did not significantly distinguish SQS and DQS subjects. The complaint of nonrestorative sleep is not a useful indicator of insomnia, despite its inclusion in all medical classifications. DQS - I and SQS + I subjects defy traditional classifications. A better understanding of sleep complaints and more accurate classifications will help physicians identify patients with insomnia and meet their needs more appropriately. PMID- 9406324 TI - Electrophysiological changes during the sleep onset period of psychophysiological insomniacs, psychiatric insomniacs, and normal sleepers. AB - The electroencephalograms (EEGs) of the sleep onset period (SOP) of psychophysiogical insomniacs. psychiatric insomniacs, and controls were compared using power spectral analysis. We predicted that psychophysiological insomniacs would show elevated cortical arousal throughout their entry into sleep. Electroencephalograms, electrooculograms (EOGs), and electromyograms (EMGs) were recorded for two consecutive nights. Power spectral analysis of EEG from the sleep onset period was performed on all standard frequency bands. Psychophysiological insomniacs had less alpha during the first part of the SOP and did not show the dramatic drop in alpha across the SOP that characterizes normal sleep. They showed less delta in the last quartile of the chronological analysis of the SOP. Psychiatric insomniacs showed lower relative beta power values overall, while psychophysiological insomniacs showed higher relative beta power values during wakefulness. This microanalysis indicates that the SOP is generally similar for psychiatric insomniacs and normal sleepers but that clear differences in the SOP of psychophysiological insomniacs exist. They have higher cortical arousal during the SOP than do psychiatric insomniacs and controls. The dramatic changes in power values in the latter two groups as sleep begins are not seen in the psychophysiological insomniacs, which may help explain the difficulty that psychophysiological insomniacs have discriminating between wakefulness and sleep. PMID- 9406325 TI - Nocturnal eating: prevalence and features in 120 insomniac referrals. AB - Pathologic nocturnal eating can be associated with a heterogeneous group of medical and psychiatric disorders. The current study was designed to evaluate the prevalence and clinical features of nocturnal eating syndrome (NES), a major subtype of pathological nocturnal eating. Conducted prospectively over an 18 month period (January 1994-June 1995), the study consisted of clinical, psychological, and polysomnographic assessments of 120 adult subjects (51 males, 69 females; mean age 42.6 years, range 18-86 years) who were either self referrals (58%) or physician referrals (42%) to our Sleep Disorders Center for insomnia complaints. Nocturnal eating with features that are typical of NES, namely compulsive feeding shortly after a mid-non-rapid eye movement (NREM) sleep awakening, in the absence of daytime eating disorders, occurred in seven subjects (five females, two males; mean age 50.8 +/- 9.5 years; mean age at onset of NES 42 years, range 18-61 years), or 5.8% of the sample. NES accounted for 44.4% of all the nocturnal eating cases observed. The data suggest that an adult, late onset variety of NES is not infrequent. Several of the clinical features of our NES patient series correspond closely to most of those observed in other descriptions of NES in the literature. Overall, the data reinforce the idea that NES is a distinct syndrome, even though some of its features overlap with sleep related eating disorders (e.g. associated with sleepwalking, restless legs syndrome, obstructive sleep apnea, etc.) and with eating disorders such as daytime binge eating. PMID- 9406326 TI - Motor overactivity and loss of motor circadian rhythm in fatal familial insomnia: an actigraphic study. AB - The 24-hour rest-activity pattern and the amount of motor activity was studied in a patient with fatal familial insomnia (FFI) by means of wrist actigraphy. During the study, the patient underwent indirect calorimetry. The 52-day recording showed severe disruption of the 24-hour rest-activity pattern with increased motor activity up to 80%. The 24-hour energy expenditure, assayed in a respiration chamber, was strikingly elevated by 60%. Chronic motor overactivity and loss of circadian rest-activity rhythm may play a role in the progressive metabolic exhaustion leading to death in FFI patients. PMID- 9406327 TI - Precise measurement of individual rapid eye movements in REM sleep of humans. AB - An automated analyzer for individual eye movements (EMs) has been developed that enables precise analyses of their incidence. Three new parameters for each EM are obtained: EM magnitude, the angle and speed of eyeball rotation, and the energy of each EM. All rapid eye movement (REM) sleep EMs from 40 nights of polysomnography for 20 healthy young men were analyzed. The mean frequency of eye movement (EM frequency) was 15.9 per minute. Compared to conventionally analyzed rapid eye movement (REM) density, EM frequency was more sensitive to differences among sleep cycles, nights, and individuals. The mean EM rotation was 6.27 +/- 0.021 degrees, the mean speed of rotation was 58.73 +/- 0.18 degrees/second, and mean energy was 525.85 +/- 3.82 degrees2/second. The distribution of changes in these new parameters differed from conventional measures across REM episodes. The conventional measures, REM episode duration, and REM density increased progressively in successive REM episodes in an ascent-to-right pattern. However, the new parameters peaked in the second, followed by relatively low values, producing an inverted V pattern. This discrepancy could indicate physiological mechanisms of EM that are not revealed in conventional measures of REM sleep intensity. PMID- 9406328 TI - Different phase relationships between EEG frequency bands during NREM and REM sleep. AB - Phase relationships between distinct frequency bands of the sleep electroencephalogram (EEG) were studied in healthy subjects using cross correlation coefficients, both over the entire night and separately for nonrapid eye movement (NREM) and rapid eye movement (REM) sleep. Over the entire night, a large positive correlation developed within high- and low-frequency bands, while a negative correlation emerged between low- and high-frequency bands, reflecting their reciprocal temporal course. More detailed analysis revealed different phase relationships during NREM and REM sleep. Findings during NREM were similar to the entire night. However, during REM, a large increase of the correlation between low- and high-frequency bands occurred compared to NREM, even with a change of sign. The results indicate the complexity of the interrelation of the different frequency bands depending on sleep state, with low- and high-frequency bands being out of phase during NREM and in phase during REM sleep. PMID- 9406330 TI - Sleep apnea and hypertension--what a mess! AB - It is unarguable that obstructive sleep apnea (OSA) causes pulsatile hypertension during sleep, but whether there is significant carryover of hypertension into waking hours is far from clear. It is perhaps more useful to consider whether OSA is related to the consequences of hypertension (e.g. stroke), since both nocturnal and daytime hypertension could be responsible for these. Furthermore, the effects of nasal continuous positive airway pressure (CPAP) on hypertension (or its consequences) must be assessed by randomized controlled studies, in exactly the same way as trials on hypotensive drugs would be carried out, before treatment is prescribed for OSA in the absence of any daytime symptoms. PMID- 9406329 TI - Contributions of the pedunculopontine region to normal and altered REM sleep. AB - The pedunculopontine (PPN) region of the upper brainstem is recognized as a critical modulator of activated behavioral states such as wakefulness and rapid eye movement (REM) sleep. The expression of REM sleep-related physiology (e.g. thalamocortical arousal, ponto-geniculate-occipital (PGO) waves, and atonia) depends upon a subpopulation of PPN neurons that release acetylcholine (ACh) to act upon muscarinic receptors (mAChRs). Serotonin's potent hyperpolarization of cholinergic PPN neurons is central to present working models of REM sleep control. A growing body of experimental evidence and clinical experience suggests that the responsiveness of the PPN region, and thereby modulation of REM sleep, involves closely adjacent glutamatergic neurons and alternate afferent neurotransmitters. Although many of these afferents are yet to be defined, dopamine-sensitive GABAergic pathways exiting the main output nuclei of the basal ganglia and adjacent forebrain nuclei appear to be the most conspicuous and the most likely to be clinically relevant. These GABAergic pathways are ideally sited to modulate the physiologic hallmarks of REM sleep differentially (e.g. atonia versus cortical activation), because each originates from a functionally unique forebrain circuit and terminates in a unique pattern upon brain stem neurons with unique membrane characteristics. Evidence is reviewed that changes in the quality, timing, and quantity of REM sleep that characterize narcolepsy, REM sleep behavior disorder, and neurodegenerative and affective disorders (depression and schizophrenia) reflect 1) changes in responsiveness of cells in the PPN region governed by these afferents; 2) increase or decrease in PPN cell number; or 3) mAChRs mediating increased responsiveness to ACh derived from the PPN. Auditory evoked potentials and acoustic startle responses provide means independent from recording sleep to assess pathophysiologies affecting the PPN and its connections and thereby complement investigations of their role in affecting daytime functions (e.g. arousal and attention). PMID- 9406331 TI - Essential hypertension and abnormal upper airway resistance during sleep. AB - The epidemiological, clinical, hereditary, biochemical, hematological, and physiological characteristics of essential hypertension (EH) and obstructive sleep apnea (OSA) are reviewed here. This extensive review shows that essential hypertension and sleep-disordered breathing--independently of whether it is OSA syndrome or upper airway resistance syndrome--share strikingly similar characteristics. The accumulated data obtained by many different researchers support the hypothesis that EH is mainly due to increased upper airway resistance during sleep. If this hypothesis is correct, treating disorders that cause increased upper airway resistance, particularly during sleep, would be an important part of the treatment of essential hypertension. PMID- 9406332 TI - Sleep-disordered breathing and hypertension: past lessons, future directions. AB - That obstructive sleep apnea syndrome is an independent risk factor for the development of hypertension was established in the 1970s, and recent works on large samples have confirmed this fact. Investigations of the mechanisms that may lead to the development of hypertension with sleep-disordered breathing will allow not only confirmation of the relationship but also creation of better treatment. There is a multigenic basis of blood pressure regulation, and genetic factors play a role in the development of sleep-disordered breathing. Genes that may have little role in the physiologic variation of blood pressure may be more important in the manifestation of pathology. And one hypothesis is that genes involved in the development of a morphotype may also have a role in the development of hypertension. Furthermore, sleep-disordered breathing may be associated with abnormal sympathetic discharge during sleep, as shown by microneurography. This mechanism may explain how a sleep disorder leads to hypertension, but impairment of vascular endothelial controls may also be involved. Investigation of vascular endothelial vasodilation as demonstrated by forearm plethysmography or the dorsal hand vein technique indicates that impairment of endothelium-dependent vasodilation during wake is associated with sleep-disordered breathing. This endothelium-dependent vasodilation appears to be more frequently impaired than the endothelium-independent vasodilation, and the former impairment can be reversed by nasal continuous positive airway pressure. These findings are supportive fo the role of sleep-disordered breathing in the development of hypertension in man. PMID- 9406333 TI - Clonazepam treatment of rhythmic movement disorders. PMID- 9406335 TI - Bibliography of recent literature in sleep research. PMID- 9406334 TI - Frequency of multiple sleep onset REM periods in "control" subjects. PMID- 9406336 TI - Rural hospital social work: views of physicians and social workers. AB - The existence of rural hospitals is financially tenuous due to the Prospective Payment system, which differentially reimburses rural facilities at lower rates than urban facilities. Physicians and hospital social workers provide services within these severely cost-conscious organizations to patients from communities, which are also aftercare resource poor. Given these multiple constraints, rural physicians' expectations of hospital social workers is integral to effective collaboration and social service provision. However, no prior investigation of these rural professionals exists. This research investigated the expectations of rural physicians and social workers regarding social work in the acute care rural hospital. The findings identify areas of disagreement and have implications for professional collaboration and education for rural hospital practice. PMID- 9406337 TI - Factors influencing priorities in hospital social work departments: a director's perspective. AB - This study compares the perceived influence of several factors, each representing a popular perspective, on priority setting in hospital social work departments: (1) leader characteristics of the director; (2) organizational characteristics of the department and hospital; and (3) the preferences of constituency groups. The authors surveyed the views of directors to ascertain influences on their allocation of resources. We find that organizational factors and the preferences of constituents are the strongest determinants of departmental priorities, with leader attributes playing a less influential role. An interesting discovery is that each factor's influence varies depending on the nature of the priority area. We conclude that all three explanations for how performance priorities are shaped a political model, a leader influence model and an institutional model-find support. The authors interpret and assess the significance of these findings to the practice of social work administration in hospitals. PMID- 9406338 TI - Discharge planning: an examination of the perceptions and recommendations for improved discharge planning at the Montreal General Hospital. AB - Hospital discharge planning is an interdisciplinary hospital-wide process that assists patients and their families to develop a feasible post-hospital plan of care. In order to facilitate an effective, smooth and rapid discharge plan for patients, health care professionals involved in discharge planning must be aware of problems and current gaps in service delivery. In order to identify and address problems and service gaps in an acute care trauma centre, the Social Work Department of the Montreal General Hospital conducted a survey of 81 clinicians via a questionnaire and interviewed 15 managers and administrators as to their perceptions and recommendations of the discharge planning process in their hospital. Internal, external and psychosocial factors were examined and three categories of discharge impediments emerged: systems, resources and patient/family issues. Recommendations for improving the discharge planning process are discussed. PMID- 9406339 TI - Social work practice with deaf clients: issues in culturally competent assessment. AB - Persons with severe hearing loss live in a unique cultural context with which social workers may not be familiar. This paper reviews the skills needed for the culturally competent social work assessment with deaf clients, including communication skills, interviewing methods, taking case and family histories and behavioral observation. PMID- 9406340 TI - Persons with traumatic brain injuries and their families: living arrangements and well-being post injury. AB - This article discusses an empirical study of 66 single, adult survivors of moderate to severe traumatic brain injury (TBI) who were either living alone or living with their parents. The researchers determined perceived levels of stress, life satisfaction, family satisfaction, and community integration within the two groups. Survivors, who were typically ten years or more post injury, and selected family members, responded to structured telephone interviews and standardized questionnaires. The findings of the study and their implications point to a need for practice interventions that will reestablish life cycle trajectories and meet developmental needs, as well as reintegrate affected individuals and their families into larger social systems. PMID- 9406341 TI - Introduction: gene vaccination, current concepts and future directions. PMID- 9406342 TI - Plasmid DNA vaccination: mechanism of antigen presentation. PMID- 9406343 TI - The development of a multivalent DNA vaccine for malaria. PMID- 9406344 TI - Genetic vaccination against tuberculosis. AB - New weapons are needed in the fight against tuberculosis. Recent research indicates that a vaccine better than BCG may be within reach. A diverse range of protein antigens can give encouragingly high levels of protective immunity in animal models when administered with adjuvants or as DNA vaccines. Accelerated arrest of bacterial multiplication followed by sustained decline in bacterial numbers are key parameters of protection and so the vaccine must target antigens produced by both actively multiplying and growth-inhibited bacteria. Consistent with this, the protective antigens have been found among secreted and stress proteins (e.g. Ag85, ESAT-6, hsp65, hsp70). Species-specific antigens are not needed, hence these remain available for diagnostic tests. Adoptive transfer of protection from vaccinated or infected mice into naive mice by transfer of purified T cells and clones shows that protection is expressed by antigen specific cytotoxic T cells that produce interferon-gamma and lyse infected macrophages. These cells are produced in response to endogenous antigen. DNA vaccination appears to be an excellent way of generating these cells and may be able to give long-lasting protection. PMID- 9406345 TI - DNA gene vaccination for HIV. PMID- 9406346 TI - DNA-based immunization against hepatitis B virus. PMID- 9406347 TI - Gene vaccination for hepatitis C. PMID- 9406348 TI - Gene immunization for allergic disorders. PMID- 9406349 TI - Immunization with DNA vaccines in early life: advantages and limitations as compared to conventional vaccines. PMID- 9406350 TI - DNA vaccines: safety and efficacy issues. AB - DNA technology has been harnessed to produce a variety of plasmid-based vaccines designed to prevent viral, bacterial and parasitic infections. The rapid adoption and implementation of this novel vaccine strategy carries with it important safety and efficacy concerns. This review will focus on whether DNA vaccines (1) are likely to induce systemic or organ-specific autoimmune disease, (2) have the potential to induce tolerance rather than immunity, and (3) are as effective in individuals with depressed immune function as they are in healthy adults. PMID- 9406351 TI - Genetic vaccines--a revolution in vaccinology? PMID- 9406352 TI - How safe are opioids in palliative care? PMID- 9406353 TI - An interview with Dr. Franco De Conno from the National Tumour Institute in Milan. Interview by F. Stiefel. PMID- 9406354 TI - Department of palliative care in Bratislava and the development of the palliative care movement in Slovakia. AB - Slovakia is a country with no tradition of home care services and a long history of regarding death and dying as taboos and therefore institutionalising them. Increased attention to palliative care issues has resulted in some important changes, to the benefit of patients in need of palliative care. These include general availability of oral slow-release forms of strong opioids (cost completely reimbursed by the insurance companies), a developing network of home care agencies, and increased attention to the needs of palliative patients, especially among oncologists and pain specialists. In February 1995 the Department of Palliative Care was established within the National Cancer Institute in Bratislava. It has 19 in-patient beds and also an out-patient clinic. Although the primary goal is the improvement of the quality of life, several approaches that can prolong life without worsening its quality are also used. These include laser destruction of intraluminal gastrointestinal tumours, insertion of intraluminal stents, brachyradiotherapy, pleurodeses, percutaneous gastrostomy, percutaneous nephrostomy, palliative chemotherapy, treatment of hypercalcaemia. In 1995 the Palliative Care Section of the Association for Study and Treatment of Pain was established, as was the first Hospice Foundation. PMID- 9406355 TI - Quality of life and supportive care. AB - Quality of life and supportive care are complementary concepts in the care of cancer patients. Neither is easy to define. Both have received increasing attention in the medical literature of recent years. From the clinical perspective, supportive care is one means toward the end of improving patients' quality of life. In order to evaluate our degree of success in this endeavour, we must agree on operational definitions of those aspects of care and its outcome we wish to study, then devise, validate and apply appropriate measures. Supportive care covers a variety of topics including symptom control, anti-infective measures, nutritional supplements and psychosocial support. The aspects of quality of life studied include physical, emotional, psychological and (less commonly) spiritual wellbeing. Symptoms influenced by the disease or its treatment are often included in the assessment. Quality of life scales have been used as outcome measures in comparing treatments, and have shown independent prognostic value. This has led several groups to examine the potential of psychosocial interventions aimed at increasing duration of survival by improving quality of life. Quality of life can and should be measured as part of the assessment of the adequacy and effectiveness of supportive care. PMID- 9406356 TI - Opioids: overview on action, interaction and toxicity. AB - The history of opioid use in briefly reviewed, and the presently accepted indications are discussed with reference to dosage, modes of administration, efficacy, duration of effect and speed of onset, and possible side effects. Physicians' fears about dependence and addiction are also touched upon. PMID- 9406357 TI - Problems with opiates in cancer pain: parenteral opioids. AB - Morphine is the preferred drug for the management of moderate-severe chronic cancer pain. The best route of administration is by mouth, because it is simple, safe, convenient, inexpensive and effective. Non-oral modes of administration should be only considered if (a) the oral route becomes unavailable or (b) there is documentation of failure of maximal doses of oral morphine and coanalgesic drugs. Recent developments have made new routes of morphine administration fashionable--in the absence of supportive pharmacokinetic or pharmacodynamic data -even when departure from established practice is not justified. It is important for clinicians to be familiar with the practicalities and problems that limit the utility of the non-oral routes, and the state of current understanding of these options will be reviewed. PMID- 9406358 TI - Organizational barriers in opioid use. AB - Despite the fact that we have both the means and the knowledge to ameliorate most forms of pain effectively, a significant number of cancer patients still experience unacceptable levels of pain. This paper sets out to explore the nature of the various organizational barriers to effective pain management. There is ample evidence to demonstrate that both physicians and nurses lack knowledge regarding modern methods of pain control. This situation not only results in poor clinical decision making, but has also spawned a number of extraordinary myths and misconceptions about the use of opioids. Such myths and misconceptions often result in significant undermedication of the patient's pain. Problems can also exist with continuity of care--the patient may be seen by a number of different physicians across a number of different health care settings where no one person is willing to take responsibility for the overall management of the patient's pain. Further fragmentation can occur due to lack of communication between the hospital and the community care setting. This problem can be compounded by incomplete and inconsistent documentation of pain. An important and often overlooked problem relating to opioid use is the existence of bureaucratic regulations governing the supply, prescription and administration of opioids in many countries world wide. There appears to be a real fear that liberalizing many of these regulations will result in an increase in illicit drug use. This paper will conclude with a discussion on ways in which the above-mentioned organizational barriers can be overcome. PMID- 9406359 TI - Pseudo-opioid-resistant pain. AB - The purpose of this article is to describe and analyse factors that result in pseudo-opioid-resistant pain. This is defined as a persistent pain experience communicated by the patient or family after prescription and initiation of opioid therapy based on empirically validated criteria. Pseudo-opioid-resistant pain can be caused by inadequate self-care or family care in relation to opioid therapy. Problems can arise in relation to communication of the pain experience, acceptance of the treatment choice and correct opioid administration. These problems may result from misconceptions or knowledge deficit, lack of motivation and lack of performance capabilities. The article systematically analyses the three categories of aetiological factors and arrives at a comprehensive explanatory model. This can be used for research purposes as well as for problem detection in clinical practice. The article includes a case report. PMID- 9406360 TI - Genetic detection: the need for psychosocial support in modern cancer prevention. AB - During recent years the gap between the rapid implementation of new technologies in cancer prevention and the slow development of a complementary psychological framework to conceptualize the transmission of genetic informations to patients has been deplored. Such a framework should include all psychological aspects surrounding the genetic consultation, reaching from the information and education of the general public to the impact of prophylactic surgery. While some of the psychological consequences of modern cancer prevention can not be fully foreseen and have first to be documented and analysed, others can easily be anticipated. The authors will try to outline a psychological framework that could help in facing potential negative effects of these beneficial preventive possibilities. PMID- 9406361 TI - Adjustment and coping by parents of children with cancer: a review of the literature. AB - Studies published since about 1980 on psychological adjustment and coping of parents of children with cancer were reviewed. First, results concerning parental adjustment in terms of psychological distress, marital distress, and family functioning were summarized. Secondly, the use of coping strategies such as social support, communication, and search for meaning were described. Thirdly, factors that influence parental adjustment to childhood cancer, such as coping strategies and illness-related and demographic variables were discussed. All studies are summarized in a review table, with information about the number of participating parents and children, the purpose, measures and major results. Difficulties in generalizing findings are possibly due to the heterogeneous group of children with cancer, the differences in reporting emotional problems by mothers and fathers, the difficulties in assessing illness-specific problems, and the diversity in the ways of assessing coping and adjustment. PMID- 9406362 TI - Peroperative teicoplanin for prevention of gram-positive infections in neutropenic patients with indwelling central venous catheters: a randomized, controlled study. AB - A prospective, randomized, open study comparing two doses of teicoplanin with no therapy administered at the time of insertion of a central venous catheter was performed in patients with hematological malignancies and in patients scheduled to undergo allogeneic or autologous stem cell transplantation. The study was designed as a group sequential study. At predetermined intervals statistical analysis was performed for the main efficacy variable, which was the number of days to treatment failure. Sixty-five patients were randomized. Three patients were judged to be not evaluable. Baseline characteristics were identical in the two groups. No differences were found in overall infections, bacteremias, gram positive infections, or local infections between the teicoplanin and control groups. Teicoplanin given at the time of insertion of central venous catheters did not reduce the risk of bacteremias or other line-associated infections. PMID- 9406363 TI - A pilot study to evaluate the feasibility of using willingness to pay as a measure of value in cancer supportive care: an assessment of amifostine cytoprotection. AB - The most commonly used method for pharmacoeconomic studies has been the cost effectiveness analysis (CEA), where the outcome is expressed as an incremental cost per unit of effectiveness (e.g. quality-adjusted life years). Although CEA is a valuable tool for identifying therapies that are more effective and less expensive, deficiencies develop when a given treatment is both more expensive and more effective. An alternative that has not been investigated in the oncology setting is the willingness-to-pay (WTP) method. In this pilot study, a WTP strategy was utilized to estimate the value that the Canadian tax-paying public puts on amifostine, a new cytoprotective agent that reduces the risk of chemotherapy-induced toxicity. The method of WTP was used within the framework of a classical cost-benefit analysis to estimate the net cost or benefit of prophylactic amifostine in patients with ovarian cancer who were receiving chemotherapy. This included direct costs for amifostine administration and hospital savings secondary to the reduced incidence of antineoplastic toxicity. A random sample of 50 Canadian tax-payers were interviewed to ascertain their maximum WTP for the new drug. The WTP survey instrument was simple to administer and easily understood by participants. Respondents stated that they would be willing to pay an average of $Can3,476 (95% confidence interval = $Can2,275 to $Can4,676) as an income tax increase to be paid over their lifetime for the value offered by the product. The benefit was then subtracted from the overall cost of amifostine ($Can3,826). This produced a net cost of $Can350 per patient (95% confidence interval = -$Can850 to $Can1,551), suggesting a situation of cost neutrality. WTP as a measure of value for oncology products is feasible and should be considered for future economic evaluations. The strategy is currently being used at this institution to determine the net societal cost or benefit of other cancer supportive care therapies, such as epoetin-alfa. PMID- 9406364 TI - Ondansetron versus a chlorpromazine and dexamethasone combination for the prevention of nausea and vomiting: a prospective, randomised study to assess efficacy, cost effectiveness and quality of life following single-fraction radiotherapy. AB - Lower hemibody radiotherapy is an effective palliative treatment for patients with wide-spread bone metastases, but is frequently associated with the unpleasant side effects of nausea and vomiting. Patients often require admission to hospital for at least an overnight stay, with its inevitable costs. This study has investigated the clinical efficacy and safety profile of ondansetron, a 5HT3 receptor antagonist, and compared it to a standard antiemetic combination, chlorpromazine and dexamethasone. Sixty-six patients were randomised to receive antiemetic prophylaxis with either oral ondansetron or a combination of chlorpromazine and dexamethasone (33 patients in each arm): 60 were treated with lower abdominal radiotherapy (8 Gy mid-plane dose) and 6 with radiotherapy to the upper lumbar spine (12.5 Gy incident dose). Patients were assessed for severity of nausea and vomiting and for whether they would use the same antiemetic again. Quality of life was assessed using the Functional Living Index Cancer (FLIC) and Functional Living Index Emesis (FLIE) quality-of-life questionnaires. A detailed cost-benefit analysis was also performed. Ondansetron scored highly as an antiemetic, being significantly better at controlling emesis on all four study days (P < 0.001) and significantly better at controlling nausea on day 1 (P < 0.001) than the standard combination of chlorpromazine and dexamethasone. Quality of life was better in the ondansetron-treated group, and ondansetron was found to be safe with no significant adverse effects. As a result, 98% of patients and investigators would use ondansetron again. Cost-benefit analysis revealed that, when complete control of emesis is the aim, ondansetron is not unduly expensive compared to the standard antiemetic regimen. As ondansetron was clearly effective in patients receiving hemibody irradiation it seems it would be prudent to adopt it for use in such patients routinely. The use of ondansetron would allow them to be treated as outpatients, with the attendant financial and psychosocial benefits of such an approach. PMID- 9406365 TI - Hypersensitivity reactions to parenteral lipid solutions. AB - In cancer patients, hypersensitivity reactions to adjunctive medications are easily mistaken for cytostatic toxicities. We report on three patients with systemic reactions (flush, dyspnea, tachycardia, hypotension, back pain) to a lipid emulsion containing long chain fatty acids (LCT). Reexposure to LCT and exposure to MCT (medium chain fatty acids) solutions of slightly different composition--no soybean lecithin used as an emulsifier--were well tolerated. These data suggest that traces of soybean proteins are the allergenic agents. Therefore, hypersensitivity to concomitant medications, including parenteral nutrition, has to be considered in oncologic patients demonstrating severe systemic reactions to intravenous therapy. PMID- 9406366 TI - Palliative treatment at home for patients with haematological disorders. AB - Supportive treatment of patients with haematological disorders mainly takes the form of transfusions of blood and platelets, and sometimes palliative chemotherapy is given. Most patients are treated in hospital or at the outpatient clinic. However, it is often difficult for the patients to arrange to come to the hospital, as they need transport by ambulance or taxi and sometimes a relative to help them. Throughout 1996 we offered such patients supportive treatment at home. A nurse was employed on the project, who was supplied with a car and a mobile telephone. Treatment was given at home. In all, 17 patients were treated, with a total of 90 blood and 40 platelet transfusions. At three visits chemotherapy was administered. No complications were seen, and the patients felt safe and content. We conclude that supportive treatment at home is safe and well accepted by patients and their relatives. In addition, the costs for transportation and hospital care of this patient group were reduced. PMID- 9406367 TI - Pathophysiology and management of subretinal hemorrhage. AB - Subretinal hemorrhage can arise from the retinal and/or choroidal circulation. Significant subretinal hemorrhage occurs in several conditions, but most commonly is associated with age-related macular degeneration, presumed ocular histoplasmosis, high myopia, retinal arterial macroaneurysm, and trauma. Released toxins, outer retinal shear forces, and a diffusion barrier created by subretinal hemorrhage all contribute to photoreceptor damage and visual loss. The use of tissue plasminogen activator and improvements in surgical instrumentation have facilitated surgical drainage and have made it a useful option in the management of selected cases. Mechanisms of subretinal hemorrhage formation, underlying etiologies, diagnostic evaluation, and the histopathology of damage are summarized. Published surgical series are reviewed and surgical advances are summarized. The value of surgically removing subretinal hemorrhages to improve visual outcome remains unestablished, because definitive studies have not been performed. Guidelines for selecting candidates for surgical intervention are proposed. PMID- 9406368 TI - Radiation therapy for choroidal melanoma. AB - Radiotherapy offers patients with malignant melanoma of the choroid an eye and a vision-sparing alternative to enucleation. The most commonly used forms of radiotherapy are ophthalmic plaque brachytherapy and charged-particle (external beam) radiotherapy. Unfortunately, after all forms of radiotherapy for choroidal melanoma many patients experience sight-limiting side effects, and an average of 16.3% of patients treated with radiotherapy subsequently require enucleation because of tumor regrowth or uncontrollable neovascular glaucoma. The severity, location, and incidence of radiation-induced complications are related to the type of radiation used, its method of delivery, amount of radiation delivered to normal ocular structures, the size and location of the tumor, as well as its response to irradiation. Current research is directed toward developing methods to reduce the amount of radiation delivered to normal structures, e.g., adding heat to radiotherapy. The true viability and metastatic potential of irradiated uveal melanoma cells has not been established, although clinical studies have reported local control of choroidal melanoma in 81-100% (mean = 92.8%) of cases. The purpose of this review is to present the world's experience with radiotherapy for choroidal melanoma, information that will contribute to patient education and informed consent. PMID- 9406369 TI - Mucosal defense of the outer eye. AB - A combination of mechanical, anatomical, immunological, and microbiological factors prevent infection of the outer eye. Mechanical and anatomical factors include the intact epithelium of the conjunctiva and cornea and the constant blinking action of the eyelids. Tear components that play a role in eye defense include lysozyme, immunoglobulins, lactoferrin, and betalysin. The normal bacterial flora of the conjunctiva may also have an inhibitory effect on the survival of more pathogenic species. The eye is linked to the common mucosal immune system, thus gaining the benefits of a system of microbial defense which is primed in the gastrointestinal tract, where a continuing large antigen load is capable of stimulating ongoing immune protection. The relative roles of the various factors contributing to prevention of eye infection remain to be fully defined. PMID- 9406370 TI - Big muscles and big nerves. AB - For 11 years, a 50-year-old woman with euthyroid Graves' disease experienced intermittent exacerbations of her orbitopathy associated with a decline in visual acuity. On each occasion, treatment with systemic corticosteroids led to prompt recovery of vision. Upon referral for consideration for orbital decompressive surgery, computed tomography and magnetic resonance imaging scanning detected bilateral optic-nerve sheath meningiomas, as well as typical findings of Graves' disease. Orbital radiation therapy led to stabilization of visual function and orbital findings, eliminating the need for systemic steroids. To our knowledge, this is the first reported case of Graves' disease associated with bilateral optic nerve sheath meningiomas. PMID- 9406371 TI - The CO2 laser in oculoplastic surgery. AB - The introduction of a new generation of carbon dioxide (CO2) lasers has permitted the development of new approaches toward certain oculoplastic disorders and procedures. The high absorption of this infrared laser by tissue water assists oculoplastic surgeons in performing incisional and excisional procedures precisely and with relatively good hemostasis. The development of new scanned continuous-wave or pulsed delivery systems has facilitated controlled tissue ablation with decreased collateral thermal injury during cutaneous resurfacing procedures. The unique characteristics of the CO2 laser mandate special attention to protection of the patient and surgical team, and careful preparation and training will help the prospective laser surgeon to successfully address the learning curve associated with this new technology. Although long-term follow-up is limited, results reported to date suggest that the CO2 laser represents an important addition to the armamentarium of the oculoplastic surgeon. PMID- 9406372 TI - Astigmatism and the toric intraocular lens and other vertex distance effects. AB - An intraocular lens with a given cylindrical power will correct a variable amount of cylinder at the spectacle plane. This variability depends not only on the vertex distance but also on the degree of ametropia. The amount of spectacle cylinder corrected will be more for myopes and less for hyperopes. Anterior corneal cylinder (as determined by the keratometer) is a more reliable way of estimating the cylinder needed at the intraocular lens plane. PMID- 9406373 TI - Use of insurance claims databases to evaluate the outcomes of ophthalmic surgery. AB - This article reviews methodological issues in research using Medicare claims data and reviews how those issues affect the interpretation of study results. Although studies using Medicare claims data can improve our knowledge of surgical outcomes, the ability to infer that one type of surgery or one surgeon is better than another is limited by such factors as bias due to inaccurate record keeping, confounding of treatment effects by other covariates, effect modification due to factors associated with treatment, and selective loss of follow-up of patients. Five studies on the outcomes of cataract surgery and capsulotomy are reviewed, providing illustrations of methodological strengths and weaknesses of this type of research. PMID- 9406374 TI - Liability issues associated with PRK and the excimer laser. AB - Given the elective nature of photorefractive keratectomy (PRK), the high expectations of patients, and misconceptions of the general public about refractive surgery, the use of the excimer laser for PRK opens the door to new liability risks for ophthalmologists and, in the comanagement environment, referring optometrists. The authors discuss informed consent, marketing, comanagement, and off-label use guidelines and protocols to help protect ophthalmologists against claims and better defend those that might arise. PMID- 9406375 TI - The wounding of Alexander the Great in Cyropolis (329 BC): the first reported case of the syndrome of transient cortical blindness? AB - I believe that the transient blindness which presented Alexander the Great after his being wounded on his head and/or his neck by a stone from a catapult during the siege of Cyropolis (329 BC) was in all probability a case of transient cortical blindness that was recognized as a special entity in the 1960s. I reached this conclusion after the comparative study of the Emperor's clinical picture provided by ancient texts, especially those of Plutarch and Quintus Curtius Rufus, with that of a modern medical bibliography. PMID- 9406376 TI - Dark adapted thresholds in children with histories of mild retinopathy of prematurity. PMID- 9406377 TI - Disruption of orientation tuning in visual cortex by artificially correlated neuronal activity. PMID- 9406378 TI - Aniseikonia: findings of a 10-year study. PMID- 9406379 TI - Characteristics of Duane's retraction syndrome at a clinic in South Africa. PMID- 9406380 TI - Isolation and molecular characterization of high-performance cellobiose fermenting spontaneous mutants of ethanologenic Escherichia coli KO11 containing the Klebsiella oxytoca casAB operon. AB - Escherichia coli KO11 was previously constructed to produce ethanol from acid hydrolysates of hemicellulose (pentoses and hexoses) by the chromosomal integration of Zymomonas mobilis genes encoding pyruvate decarboxylase (pdc) and alcohol dehydrogenase (adhB). Klebsiella oxytoca P2 was constructed in an analogous fashion for the simultaneous saccharification and fermentation of cellulose and contains PTS enzymes for cellobiose. In this study, KO11 was further engineered for the fermentation of cellulose by adding the K. oxytoca casAB genes encoding Enzyme IIcellobiose and phospho-beta-glucosidase. Although the two K. oxytoca genes were well expressed in cloning hosts such as DH5 alpha, both were expressed poorly in E. coli KO11, a derivative of E. coli B. Spontaneous mutants which exhibited more than 15-fold-higher specific activities for cellobiose metabolism were isolated. The mutations of these mutants resided in the plasmid rather than the host. Three mutants were characterized by sequence analysis. All contained similar internal deletions which eliminated the casAB promoter and operator regions and placed the lacZ Shine-Dalgarno region immediately upstream from the casA Shine-Dalgarno region. KO11 harboring mutant plasmids (pLOI1908, pLOI1909, or pLOI1910) rapidly fermented cellobiose to ethanol, and the yield was more than 90% of the theoretical yield. Two of these strains were used with commercial cellulase to ferment mixed-waste office paper to ethanol. PMID- 9406381 TI - The faeA genes from Aspergillus niger and Aspergillus tubingensis encode ferulic acid esterases involved in degradation of complex cell wall polysaccharides. AB - We report the cloning and characterization of a gene encoding a ferulic acid esterase, faeA, from Aspergillus niger and Aspergillus tubingensis. The A. niger and A. tubingensis genes have a high degree of sequence identity and contain one conserved intron. The gene product, FAEA, was overexpressed in wild-type A. tubingensis and a protease-deficient A. niger mutant. Overexpression of both genes in wild-type A. tubingensis and an A. niger protease-deficient mutant showed that the A. tubingensis gene product is more sensitive to degradation than the equivalent gene product from A. niger. FAEA from A. niger was identical to A. niger FAE-III (C. B. Faulds and G. Williamson, Microbiology 140:779-787, 1994), as assessed by molecular mass, pH and temperature optima, pI, N-terminal sequence, and activity on methyl ferulate. The faeA gene was induced by growth on wheat arabinoxylan and sugar beet pectin, and its gene product (FAEA) released ferulic acid from wheat arabinoxylan. The rate of release was enhanced by the presence of a xylanase. FAEA also hydrolyzed smaller amounts of ferulic acid from sugar beet pectin, but the rate was hardly affected by addition of an endo-pectin lyase. PMID- 9406382 TI - Frequency of formation of chimeric molecules as a consequence of PCR coamplification of 16S rRNA genes from mixed bacterial genomes. AB - PCR is routinely used in amplification and cloning of rRNA genes from environmental DNA samples for studies of microbial community structure and identification of novel organisms. There have been concerns about generation of chimeric sequences as a consequence of PCR coamplification of highly conserved genes, because such sequences may lead to reports of nonexistent organisms. To quantify the frequency of chimeric molecule formation, mixed genomic DNAs from eight actinomycete species whose 16S rRNA sequences had been determined were used for PCR coamplification of 16S rRNA genes. A large number of cloned 16S ribosomal DNAs were examined by sequence analysis, and chimeric molecules were identified by multiple-sequence alignment with reference species. Here, we report that the level of occurrence of chimeric sequences after 30 cycles of PCR amplification was 32%. We also show that PCR-induced chimeras were formed between different rRNA gene copies from the same organism. Because of the wide use of PCR for direct isolation of 16S rRNA sequences from environmental DNA to assess microbial diversity, the extent of chimeric molecule formation deserves serious attention. PMID- 9406383 TI - Synthesis of optically active amino acids from alpha-keto acids with Escherichia coli cells expressing heterologous genes. AB - We describe a simple method for enzymatic synthesis of L and D amino acids from alpha-keto acids with Escherichia coli cells which express heterologous genes. L amino acids were produced with thermostable L-amino acid dehydrogenase and formate dehydrogenase (FDH) from alpha-keto acids and ammonium formate with only an intracellular pool of NAD+ for the regeneration of NADH. We constructed plasmids containing, in addition to the FDH gene, the genes for amino acid dehydrogenases, including i.e., leucine dehydrogenase, alanine dehydrogenase, and phenylalanine dehydrogenase. L-Leucine, L-valine, L-norvaline, L-methionine, L phenylalanine, and L-tyrosine were synthesized with the recombinant E. coli cells with high chemical yields (> 80%) and high optical yields (up to 100% enantiomeric excess). Stereospecific conversion of various alpha-keto acids to D amino acids was also examined with recombinant E. coli cells containing a plasmid coding for the four heterologous genes of the thermostable enzymes D-amino acid aminotransferase, alanine racemase, L-alanine dehydrogenase, and FDH. Optically pure D enantiomers of glutamate and leucine were obtained. PMID- 9406386 TI - Ribotyping to compare Fusobacterium necrophorum isolates from bovine liver abscesses, ruminal walls, and ruminal contents. AB - Restriction fragment length polymorphism analysis of rRNA genes was employed to genetically compare Fusobacterium necrophorum subsp. necrophorum and F. necrophorum subsp. funduliforme isolates from multiple abscesses of the same liver and isolates from liver abscesses, the ruminal wall, and ruminal contents from the same animal. Four livers with multiple abscesses and samples of ruminal contents, ruminal walls, and liver abscesses were collected from 11 cattle at slaughter. F. necrophorum was isolated from all liver abscesses, nine ruminal walls, and six ruminal content samples. Chromosomal DNA of the isolates was extracted and single or double digested with restriction endonucleases (EcoRI, EcoRV, SalI, and HaeIII); then restriction fragments were hybridized with a digoxigenin-labeled cDNA probe transcribed from a mixture of 16S and 23S rRNAs from Escherichia coli. EcoRI alone or in combination with EcoRV yielded the most discriminating ribopatterns for comparison. Within the subspecies multiple isolates from the same liver were indistinguishable based on the ribopattern obtained with EcoRI. The hybridization patterns of liver abscess isolates were concordant with those of the corresponding isolates from ruminal walls in eight of nine sets of samples. None of the six ruminal content isolates matched either the liver abscess isolates or the ruminal wall isolates. The genetic similarity between the isolates from liver abscesses and ruminal walls supports the hypothesis that F. necrophorum isolates of liver abscesses originate from the rumen. PMID- 9406385 TI - Molecular cloning and characterization of a novel mosquitocidal protein gene from Bacillus thuringiensis subsp. fukuokaensis. AB - A novel mosquitocidal protein gene, cry20Aa, was cloned from Bacillus thuringiensis subsp. fukuokaensis (H-3a: 3d: 3e). The gene product, Cry20Aa, was naturally truncated and had a molecular mass of 86,138 Da. The Cry20Aa protein possessed five conserved sequence blocks, as do most other insecticidal Cry toxins. However, an amino acid comparison of Cry20Aa with other mosquitocidal toxins, including Cry4A, Cry4B, Cry10A, Cry11A, and Cry11B, demonstrated that Cry20Aa was quite different from other toxins except for the conserved blocks. The N terminus of Cry20Aa was, however, homologous to the N termini of Cry4A and Cry10A. Interestingly, an inverted repeat (IR1) sequence in the open reading frame of the cry20Aa gene caused incomplete expression of Cry20Aa. When this internal IR1 sequence was altered with no change of amino acid sequence, acrystalliferous B. thuringiensis cells transformed with cry20Aa gene dramatically produced crystal inclusions. However, the intact 86-kDa Cry20Aa protein is highly labile, and it is rapidly degraded to polypeptides of 56 and 43 kDa. To increase expression of the cry20Aa gene, the p20 chaperonelike protein and the cyt1Aa promoter were utilized. While p20 did not increase Cry20Aa expression or stability, chimeric constructs in which the cry20Aa gene was under control of the cyt1Aa promoter overexpressed the Cry20Aa protein in acrystalliferous B. thuringiensis. The expressed Cry20Aa protein showed larvicidal activity against Aedes aegypti and Culex quinquefasciatus. However, the mosquitocidal activity was low, probably due to rapid proteolysis to inactive 56- and 43-kDa proteins. PMID- 9406387 TI - Genetic evidence for nonrandom sorting of mitochondria in the basidiomycete Agrocybe aegerita. AB - We studied mitochondrial transmission in the homobasidiomycete Agrocybe aegerita during plasmogamy, vegetative growth, and basidiocarp differentiation. Plasmogamy between homokaryons from progeny of three wild-type strains resulted in bidirectional nuclear migration, and the dikaryotization speed was dependent on the nuclear genotype of the recipient homokaryon. Little mitochondrial migration accompanied the nuclear migration. A total of 75% of the dikaryons from the fusion lines had both parental mitochondrial haplotypes (mixed dikaryons), and 25% had only a single haplotype (homoplasmic dikaryons); with some matings, there was a strong bias in favor of one parental haplotype. We demonstrated the heteroplasmic nature of mixed dikaryons by (i) isolating and subculturing apical cells in micromanipulation experiments and (ii) identifying recombinant mitochondrial genomes. This heteroplasmy is consistent with the previously reported suggestion that there is recombination between mitochondrial alleles in A. aegerita. Conversion of heteroplasmons into homoplasmons occurred (i) during long-term storage, (ii) in mycelia regenerated from isolated apical cells, and (iii) during basidiocarp differentiation. Homokaryons that readily accepted foreign nuclei were the most efficient homokaryons in maintaining their mitochondrial haplotype during plasmogamy, long-term storage, and basidiocarp differentiation. This suggests that the mechanism responsible for the nonrandom retention or elimination of a given haplotype may be related to the nuclear genotype or the mitochondrial haplotype or both. PMID- 9406388 TI - Conjugative plasmids isolated from bacteria in marine environments show various degrees of homology to each other and are not closely related to well characterized plasmids. AB - Mercury resistance plasmids were exogenously isolated, i.e., recovered after transfer to a model recipient bacterium, from marine air-water interface, bulk water, and biofilm communities during incubation in artificial seawater without added nutrients. Ninety-five plasmids from different environments were classified by restriction endonuclease digestion, and 12 different structural plasmid groups were revealed. The plasmid types isolated from different habitats and from different sampling occasions showed little similarity to each other based on their restriction endonuclease patterns, indicating high variation and possibly a low transfer between microhabitats and/or a different composition of the microbial communities at different sites and times. With another approach in which probes derived from one of the isolated plasmids and a mercury resistance (mer) probe from Tn501 were used, similarities between plasmids from several different groups were found. The plasmids were further tested for their incompatibility by use of the collection of inc/rep probes (B/O, com9, FI, FII, HI1, HI2, I1, L/M, N, P, Q, U, W, Y) described by Couturier et al. (M. F. Couturier, P. Bex, L. Bergquist, and W. K. Maas, Microbiol. Rev. 52:375-395, 1988). Hybridizations did not reveal any identity between the 12 plasmid groups and any of the inc/rep probes tested. The results indicate that plasmids isolated from different marine habitats have replication and/or incompatibility systems that are different from the well-characterized plasmids that are commonly used in plasmid biology. This shows the need for the use of more relevant plasmids in studies of plasmid activity in the environment and development of new inc/rep probes for their characterization. PMID- 9406389 TI - The ammonia monooxygenase structural gene amoA as a functional marker: molecular fine-scale analysis of natural ammonia-oxidizing populations. AB - The naturally occurring genetic heterogeneity of autotrophic ammonia-oxidizing populations belonging to the beta subclass of the Proteobacteria was studied by using a newly developed PCR-based assay targeting a partial stretch of the gene which encodes the active-site polypeptide of ammonia monooxygenase (amoA). The PCR yielded a specific 491-bp fragment with all of the nitrifiers tested, but not with the homologous stretch of the particulate methane monooxygenase, a key enzyme of methane-oxidizing bacteria. The assay also specifically detected amoA in DNA extracted from various aquatic and terrestrial environments. The resulting PCR products retrieved from rice roots, activated sludge, a freshwater sample, and an enrichment culture were used for the generation of amoA gene libraries. No false positives were detected in a set of 47 randomly selected clone sequences that were analyzed further. The majority of the environmental sequences retrieved from rice roots and activated sludge grouped within the phylogenetic radiation defined by cultured strains of the genera Nitrosomonas and Nitrosospira. The comparative analysis identified members of both of these genera in activated sludge; however, only Nitrosospira-like sequences with very similar amino acid patterns were found on rice roots. Further differentiation of these molecular isolates was clearly possible on the nucleic acid level due to the accumulation of synonymous mutations, suggesting that several closely related but distinct Nitrosospira-like populations are the main colonizers of the rhizosphere of rice. Each of the amoA gene libraries obtained from the freshwater sample and the enrichment culture was dominated by a novel lineage that shared a branch with the Nitrosospira cluster but could not be assigned to any of the known pure cultures. Our data suggest that amoA represents a very powerful molecular tool for analyzing indigenous ammonia-oxidizing communities due to (i) its specificity, (ii) its fine-scale resolution of closely related populations, and (iii) the fact that a functional trait rather than a phylogenetic trait is detected. PMID- 9406390 TI - Identification and molecular genetic analysis of multiple loci contributing to high-level tellurite resistance in Rhodobacter sphaeroides 2.4.1. AB - The ability of the facultative photoheterotroph Rhodobacter sphaeroides to tolerate and reduce high levels of tellurite in addition to at least 10 other rare earth metal oxides and oxyanions has considerable potential for detoxification and bioremediation of contaminated environments. We report the identification and characterization of two loci involved in high-level tellurite resistance. The first locus contains four genes, two of which, trgAB, confer increased tellurite resistance when introduced into the related bacterium Paracoccus denitrificans. The trgAB-derived products display no significant homology to known proteins, but both are likely to be membrane-associated proteins. Immediately downstream of trgB, the cysK (cysteine synthase) and orf323 genes were identified. Disruption of the cysK gene resulted in decreased tellurite resistance in R. sphaeroides, confirming earlier observations on the importance of cysteine metabolism for high-level tellurite resistance. The second locus identified is represented by the telA gene, which is separated from trgAB by 115 kb. The telA gene product is 65% similar to the product of the klaB (telA) gene from the tellurite-resistance-encoding kilA operon from plasmid RK2. The genes immediately linked to the R. sphaeroides telA gene have no similarity to other components of the kilA operon. R. sphaeroides telA could not functionally substitute for the plasmid RK2 telA gene, indicating substantial functional divergence between the two gene products. However, inactivation of R. sphaeroides telA resulted in a significant decrease in tellurite resistance compared to the wild-type strain. Both cysK and telA null mutations readily gave rise to suppressors, suggesting that the phenomenon of high-level tellurite resistance in R. sphaeroides is complex and other, as yet uncharacterized, loci may be involved. PMID- 9406391 TI - Two cellulases, CelA and CelC, from the polycentric anaerobic fungus Orpinomyces strain PC-2 contain N-terminal docking domains for a cellulase-hemicellulase complex. AB - Two cDNAs encoding two cellulases, CelA and CelC, were isolated from a cDNA library of the polycentric anaerobic fungus Orpinomyces sp. strain PC-2 constructed in Escherichia coli. Nucleotide sequencing revealed that the celA cDNA (1,558 bp) and celC cDNA (1,628 bp) had open reading frames encoding polypeptides of 459 (CelA) and 449 (CelC) amino acids, respectively. The two cDNAs were 76.9 and 67.7% identical at the nucleotide and amino acid levels, respectively. Analysis of the deduced amino acid sequences showed that starting from the N termini, both CelA and CelC had signal peptides, which were followed by noncatalytic repeated peptide domains (NCRPD) containing two repeated sequences of 33 to 40 amino acid residues functioning as docking domains. The NCRPDs and the catalytic domains were separated by linker sequences. The NCRPDs were homologous to those found in several hydrolases of anaerobic fungi, whereas the catalytic domains were homologous to the catalytic domains of fungal cellobiohydrolases and bacterial endoglucanases. The linker sequence of CelA contained predominantly glutamine and proline residues, while that of CelC contained mainly threonine residues. CelA and CelC did not have a typical cellulose binding domain (CBD). CelA and CelC expressed in E. coli rapidly decreased the viscosity of carboxymethyl cellulose (CMC), indicating that there was endoglucanase activity. In addition, they produced cellobiose from CMC, acid swollen cellulose, and cellotetraose, suggesting that they had cellobiohydrolase activity. The optimal activity conditions with CMC as the substrate were pH 4.3 to 6.8 and 50 degrees C for CelA and pH 4.6 to 7.0 and 40 degrees C for CelC. Despite the lack of a CBD, CelC displayed a high affinity for microcrystalline cellulose, whereas CelA did not. PMID- 9406392 TI - Phylogenetic diversity of Archaea in sediment samples from a coastal salt marsh. AB - The Archaea present in salt marsh sediment samples from a tidal creek and from an adjacent area of vegetative marshland, both of which showed active methanogenesis and sulfate reduction, were sampled by using 16S rRNA gene libraries created with Archaea-specific primers. None of the sequences were the same as reference sequences from cultured taxa, although some were closely related to sequences from methanogens previously isolated from marine sediments. A wide range of Euryarchaeota sequences were recovered, but no sequences from Methanococcus, Methanobacterium, or the Crenarchaeota were recovered. Clusters of closely related sequences were common and generally contained sequences from both sites, suggesting that some related organisms were present in both samples. Recovery of sequences closely related to those of methanogens such as Methanococcoides and Methanolobus, which can use substrates other than hydrogen, provides support for published hypotheses that such methanogens are probably important in sulfate-rich sediments and identifies some likely candidates. Sequences closely related to those of methanogens such as Methanoculleus and Methanogenium, which are capable of using hydrogen, were also discovered, in agreement with previous inhibitor and process measurements suggesting that these taxa are present at low levels of activity. More surprisingly, we recovered a variety of sequences closely related to those from different halophilic Archaea and a cluster of divergent sequences specifically related to the marine group II archaeal sequences recently shown by PCR and probing to have a cosmopolitan distribution in marine samples. PMID- 9406393 TI - Genetic diversity of rhizobial symbionts isolated from legume species within the genera Astragalus, Oxytropis, and Onobrychis. AB - The genetic diversity of 44 rhizobial isolates from Astragalus, Oxytropis, and Onobrychis spp. originating from different geographic locations was evaluated by mapped restriction site polymorphism (MRSP) analysis of 16S rRNA genes and by PCR DNA fingerprinting with repetitive sequences (REP-PCR). A comparison of tree topologies of reference strains constructed with data obtained by MRSP and by 16S rRNA gene sequence analyses showed that the topologies were in good agreement, indicating that the MSRP approach results in reasonable estimates of rhizobial phylogeny. The isolates were distributed into 14 distinct 16S rRNA gene types clustering into three major groups which corresponded with three of the genera within the legume symbionts. Most of the isolates were within the genus Mesorhizobium. Five were identified with different genomic species nodulating Lotus spp. and Cicer arietinum. Three Astragalus isolates were classified as Bradyrhizobium, one being similar to Bradyrhizobium elkanii and another being similar to Bradyrhizobium japonicum. Six of the isolates were related to species within the genus Rhizobium. Two were similar to Rhizobium leguminosarum, and the remainder were identified as Rhizobium gallicum. DNA fingerprinting by REP-PCR revealed a high level of diversity within single 16S ribosomal DNA types. The 44 isolates were distributed into 34 REP groups. Rhizobial classification at the genus and probably also the species levels was independent of geographic origin and host plant affinity. PMID- 9406394 TI - Production of poly(3-hydroxybutyrate) by fed-batch culture of filamentation suppressed recombinant Escherichia coli. AB - Recombinant Escherichia coli XL1-Blue harboring a high-copy-number plasmid containing the Alcaligenes eutrophus polyhydroxyalkanoate synthesis genes could efficiently synthesize poly(3-hydroxybutyrate) (PHB) in a complex medium containing yeast extract and tryptone but not in a defined medium. One of the reasons for the reduced PHB production in a defined medium was thought to be severe filamentation of cells in this medium. By overexpressing an essential cell division protein, FtsZ, in recombinant E. coli producing PHB, filamentation could be suppressed and PHB could be efficiently produced in a defined medium. A high PHB concentration of 149 g/liter, with high productivity of 3.4 g of PHB/liter/h, could be obtained by the pH-stat fed-batch culture of the filamentation suppressed recombinant E. coli in a defined medium. It was also found that insufficient oxygen supply at a dissolved oxygen concentration (DOC) of 1 to 3% of air saturation during active PHB synthesis phase did not negatively affect PHB production. By growing cells to the concentration of 110 g/liter and then controlling the DOC in the range of 1 to 3% of air saturation, a PHB concentration of 157 g/liter and PHB productivity of 3.2 g of PHB/liter/h were obtained. For the scale-up studies, fed-batch culture was carried out in a 50 liter stirred tank fermentor, in which the DOC decreased to zero when cell concentration reached 50 g/liter. However, a relatively high PHB concentration of 101 g/liter and PHB productivity of 2.8 g of PHB/liter/h could still be obtained, which demonstrated the possibility of industrial production of PHB in a defined medium by employing the filamentation-suppressed recombinant E. coli. PMID- 9406396 TI - Isolation and characterization of novel iron-oxidizing bacteria that grow at circumneutral pH. AB - A gel-stabilized gradient method that employed opposing gradients of Fe2+ and O2 was used to isolate and characterize two new Fe-oxidizing bacteria from a neutral pH, Fe(2+)-containing groundwater in Michigan. Two separate enrichment cultures were obtained, and in each the cells grew in a distinct, rust-colored band in the gel at the oxic-anoxic interface. The cells were tightly associated with the ferric hydroxides. Repeated serial dilutions of both enrichments resulted in the isolation of two axenic strains, ES-1 and ES-2. The cultures were judged pure based on (i) growth from single colonies in tubes at dilutions of 10(-7) (ES-2) (ES-2) and 10(-8) (ES-1); (ii) uniform cell morphologies, i.e., ES-1 was a motile long thin, bent, or S-shaped rod and ES-2 was a shorter curved rod; and (iii) no growth on a heterotrophic medium. Strain ES-1 grew to a density of 10(8) cells/ml on FeS with a doubling time of 8 h. Strain ES-2 grew to a density of 5 x 10(7) cells/ml with a doubling time of 12.5 h. Both strains also grew on FeCO3. Neither strain grew without Fe2+, nor did they grow with glucose, pyruvate, acetate, Mn, or H2S as an electron donor. Studies with an oxygen microelectrode revealed that both strains grew at the oxic-anoxic interface of the gradients and tracked the O2 minima when subjected to higher O2 concentrations, suggesting they are microaerobes. Phylogenetically the two strains formed a novel lineage within the gamma Proteobacteria. They were very closely related to each other and were equally closely related to PVB OTU 1, a phylotype obtained from an iron-rich hydrothermal vent system at the Loihi Seamount in the Pacific Ocean, and SPB OTU 1, a phylotype obtained from permafrost soil in Siberia. Their closest cultivated relative was Stenotrophomonas maltophilia. In total, this evidence suggests ES-1 and ES-2 are members of a previously untapped group of putatively lithotrophic, unicellular iron-oxidizing bacteria. PMID- 9406395 TI - Electrostatic interactions, but not the YGNGV consensus motif, govern the binding of pediocin PA-1 and its fragments to phospholipid vesicles. AB - The purpose of this study was to characterize in detail the binding of pediocin PA-1 and its fragments to target membranes by using tryptophan fluorescence as a probe. Based on a three-dimensional model (Y. Chen, R. Shapira, M. Eisenstein, and T. J. Montville, Appl. Environ. Microbiol. 63:524-531, 1997), four synthetic N-terminal pediocin fragments were selected to study the mechanism of the initial step by which the bacteriocin associates with membranes. Binding of pediocin PA-1 to vesicles of phosphatidylglycerol, the major component of Listeria membranes, caused an increase in the intrinsic tryptophan fluorescence intensity with a blue shift of the emission maximum. The Stern-Volmer constants for acrylamide quenching of the fluorescence of pediocin PA-1 in buffer and in the lipid vesicles were 8.83 +/- 0.42 and 3.53 +/- 0.67 M-1, respectively, suggesting that the tryptophan residues inserted into the hydrophobic core of the lipid bilayer. The synthetic pediocin fragments bound strongly to the lipid vesicles when a patch of positively charged amino acid residues (K-11 and H-12) was present but bound weakly when this patch was mutated out. Quantitative comparison of changes in tryptophan fluorescence parameters, as well as the dissociation constants for pediocin PA-1 and its fragments, revealed that the relative affinity to the lipid vesicles paralleled the net positive charge in the peptide. The relative affinity for the fragment containing the YGNGV consensus motif was 10-fold lower than that for the fragment containing the positive patch. Furthermore, changing the pH from 6.0 to 8.0 decreased binding of the fragments containing the positive patch, probably due to deprotonation of His residues. These results demonstrate that electrostatic interactions, but not the YGNGV motif, govern pediocin binding to the target membrane. PMID- 9406397 TI - Partial purification and characterization of manganese-oxidizing factors of Pseudomonas fluorescens GB-1. AB - The Mn(2+)-oxidizing bacterium Pseudomonas fluorescens GB-1 deposits Mn oxide around the cell. During growth of a culture, the Mn(2+)-oxidizing activity of the cells first appeared in the early stationary growth phase. It depended on the O2 concentration in the culture during the late logarithmic growth phase. Maximal activity was observed at an oxygen concentration of 26% saturation. The activity could be recovered in cell extracts and was proportional to the protein concentration in the cell extracts. The specific activity was increased 125-fold by ammonium sulfate precipitation followed by reversed-phase and gel filtration column chromatographies. The activity of the partly purified Mn(2+)-oxidizing preparation had a pH optimum of circa 7 and a temperature optimum of 35 degrees C and was lost by heating. The Mn(2+)-oxidizing activity was sensitive to NaN3 and HgCl2. It was inhibited by KCN, EDTA, Tris, and o-phenanthroline. Although most data indicated the involvement of protein in Mn2+ oxidation, the activity was slightly stimulated by sodium dodecyl sulfate at a low concentration and by treatment with pronase and V8 protease. By polyacrylamide gel electrophoresis, two Mn(2+)-oxidizing factors with estimated molecular weights of 180,000 and 250,000 were detected. PMID- 9406398 TI - Lysine-overproducing mutants of Saccharomyces cerevisiae baker's yeast isolated in continuous culture. AB - Saccharomyces cerevisiae baker's yeast mutants which produce 3 to 17 times as much lysine as the wild type, depending on the nitrogen source, have been selected. The baker's yeast strain was growth in a pH-regulated chemostat in minimal medium with proline as the nitrogen source, supplemented with increasing concentrations of the toxic analog of the lysine S-2-aminoethyl-L-cysteine (AEC). The lysine-overproducing mutants, which were isolated as AEC-resistant mutants, were also resistant to high external concentrations of lysine and to alpha aminoadipate and seemed to be affected in the lysine biosynthetic pathway but not in the biosynthetic pathways of other amino acids. Lysine overproduction by one of the mutants seemed to be due to, at least, the loss of repression of the homocitrate synthase encoded by the LYS20 gene. The mutant grew slower than the wild type, and its dough-raising capacity was reduced in in vitro assays, probably due to the toxic effects of lysine accumulation or of an intermediate produced in the pathway. This mutant can be added as a food supplement to enrich the nutritive qualities of bakery products, and its resistance to alpha aminoadipate, AEC, and lysine can be used as a dominant marker. PMID- 9406399 TI - Purification and characterization of a cephalosporin esterase from Rhodosporidium toruloides. AB - A novel cephalosporin esterase (EC 3.1.1.41) from Rhodosporidium toruloides was purified to gel electrophoretic homogeneity. The enzyme is a glycoprotein with a molecular mass of 80 kDa. Upon deglycosylation, several forms of the enzyme were observed with a molecular mass range between 60 and 66 kDa. The isoelectric point of the enzyme is approximately 5.6, with the pH optimum for activity occurring at 6.0. The optimal activity of the enzyme occurred at 25 degrees C, with the enzyme rapidly losing activity at temperatures above 25 degrees C. The enzyme deacetylated a variety of cephalosporin derivatives, including cephalosporin C; the Km for this substrate is 51.8 mM, and the Vmax is 7.9 mumol/min/mg. In addition to cephalosporins, the enzyme hydrolyzed short-chain p-nitrophenyl esters, with the activity decreasing with increasing ester chain length. The enzyme also has the ability to acetylate desacetyl cephalosporins in high yields under mild conditions in the presence of various acetyl donors. A comparison of the physical properties of the esterase with those of other well-characterized cephalosporin esterases indicates that the enzyme is unique in this class. PMID- 9406400 TI - Growth reduction of Listeria spp. caused by undefined industrial red smear cheese cultures and bacteriocin-producing Brevibacterium lines as evaluated in situ on soft cheese. AB - The undefined microbial floras derived from the surface of ripe cheese which are used for the ripening of commercial red smear cheeses have a strong impact on the growth of Listeria spp. In some cases, these microbial consortia inhibit Listeria almost completely. From such undefined industrial cheese-ripening floras, linocin M18-producing (lin+) (N. Valdes-Stauber and S. Scherer, Appl. Environ. Microbiol. 60:3809-3814, 1994) and -nonproducing Brevibacterium linens strains were isolated and used as single-strain starter cultures on model red smear cheeses to evaluate their potential inhibitory effects on Listeria strains in situ. On cheeses ripened with lin+ strains, a growth reduction of L. ivanovii and L. monocytogenes of 1 to 2 log units was observed compared to cheeses ripened with lin strains. Linocin M18 activity was detected in cheeses ripened with lin+ strains but was not found in those ripened with lin strains. We suggest that production of linocin M18 contributes to the growth reduction of Listeria observed on model red smear cheeses but is unsufficient to explain the almost complete inhibition of Listeria caused by some undefined microbial floras derived from the surface of ripe cheeses. PMID- 9406401 TI - Temperature determines the pattern of anaerobic microbial dechlorination of Aroclor 1260 primed by 2,3,4,6-tetrachlorobiphenyl in Woods Pond sediment. AB - Reductive dechlorination of the Aroclor 1260 residue in Woods Pond (Lenox, Mass.) sediment samples was investigated for a year at incubation temperatures from 4 to 66 degrees C. Sediment slurries were incubated anaerobically with and without 2,3,4,6-tetrachlorobiphenyl (2346-CB; 350 microM) as a primer for dechlorination of the Aroclor 1260 residue. Dechlorination of the Aroclor residue occurred only in live samples primed with 2346-CB and only at 8 to 34 degrees C and 50 to 60 degrees C. The extent and pattern of polychlorinated biphenyl (PCB) dechlorination were temperature dependent. At 8 to 34 degrees C, the dechlorination resulted in 28 to 65% decreases of the hexathrough nonachlorobiphenyls and corresponding increases in the tri- and tetrachlorobiphenyls. At 12 to 30 degrees C, 30 to 40% of the hexa- through nonachlorobiphenyls were dechlorinated in just 3 months. The optimal temperature for overall chlorine removal was 20 to 27 degrees C. We observed four different microbial dechlorination processes with different but partially overlapping temperature ranges, i.e., Process N (flanked meta dechlorination) at 8 to 30 degrees C, Process P (flanked para dechlorination) at 12 to 34 degrees C, Process LP (unflanked para dechlorination) at 18 to 30 degrees C, and Process T (a very restricted meta dechlorination of specific hepta- and octachlorobiphenyls) at 50 to 60 degrees C. These temperature ranges should aid in the development of strategies for the enrichment and isolation of the microorganisms responsible for each dechlorination process. The incubation temperature determined the relative dominance of the four PCB dechlorination processes and the extent and products of dechlorination. Hence, understanding the effects of temperature on PCB dechlorination at contaminated sites should assist in predicting the environmental fate of PCBs or planning bioremediation strategies at those sites. PMID- 9406402 TI - Two anaerobic polychlorinated biphenyl-dehalogenating enrichments that exhibit different para-dechlorination specificities. AB - Two anaerobic polychlorinated biphenyl (PCB)-dechlorinating enrichments with distinct substrate specificities were obtained: a 2,3,4,6-tetrachlorobiphenyl (2346-CB) para-dechlorinating enrichment derived from Aroclor 1260-contaminated Woods Pond (Lenox, Mass.) sediment and a 2,4,6-trichlorobiphenyl (246-CB) unflanked para-dechlorinating enrichment derived from PCB-free Sandy Creek Nature Center (Athens, Ga.) sediment. The enrichments have been successfully transferred to autoclaved soil slurries over 20 times by using 300 to 350 microM 2346-CB or 246-CB. Both enrichments required soil for successful transfer of dechlorination activity. The 2346-CB enrichment para dehalogenated, in the absence or presence of 2346-CB, only 4 of 25 tested para halogen-containing congeners: 234-CB, 2345 CB, 2346-CB, and 2,4,6-tribromobiphenyl (246-BrB). In the presence of 246-CB, the 246-CB enrichment para dehalogenated 23 of the 25 tested congeners. However, only three congeners (34-CB, 2346-CB, and 246-BrB) were dehalogenated in the absence of 246-CB, indicating that these specific congeners initiate dehalogenation in this enrichment culture. The addition of the 2346-CB (para)-dechlorinating enrichment did not further stimulate the 2346-CB-primed dechlorination of the Aroclor 1260 residue in Woods Pond sediment samples. Compared to the addition of the primer 246-CB or the 246-CB unflanked para-dechlorinating enrichment alone, the addition of both 246-CB (300 microM) and the 246-CB enrichment stimulated the unflanked para dechlorination of the Aroclor 1260 residue in Woods Pond sediments. These results indicate that the two enrichments contain different PCB dechlorinating organisms, each with high substrate specificities. Furthermore, bioaugmentation with the enrichment alone did not stimulate the desired dechlorination in PCB-contaminated Woods Pond sediment. PMID- 9406403 TI - Mutational analysis of pcpA and its role in pentachlorophenol degradation by Sphingomonas (Flavobacterium) chlorophenolica ATCC 39723. AB - Sphingomonas (Flavobacterium) chlorophenolica ATCC 39723 degrades pentachlorophenol (PCP) through a catabolic pathway encoded by multiple genes. One gene required for PCP degradation is pcpA, which encodes information for a 30 kDa polypeptide, PcpA, found in the periplasm of the bacterium. The biological role of PcpA has remained unknown. We disrupted pcpA by replacing it with a defective copy through homologous recombination. The pcpA recombinant, mutant strains accumulated 2,6-dichlorohydroquinone (2,6-DiCH) as a metabolite of PCP. This work confirms that pcpA is essential for degradation of PCP by S. chlorophenolica ATCC 39723 and suggests that it encodes a protein involved in hydrolytic dehalogenation of 2,6-DiCH, an already established primary metabolite of the PCP catabolic pathway. PMID- 9406405 TI - Detection of stable pre-rRNA in toxigenic Pseudo-nitzschia species. AB - Nucleotide sequence analysis of ribosomal DNA (rDNA) spacer regions is useful for taxonomic comparisons of closely related microorganisms. These regions have been less useful for routine microbial identification and detection, partly because rRNA precursors (pre-rRNAs) in microbial cells are assumed to be too labile to be detectable by high-throughput probe hybridization methods. We characterized the sequence diversity and physiological stability of pre-rRNA in the toxigenic marine diatoms Pseudo-nitzschia australis, P. multiseries, and P. pungens. As with nucleotide sequences of the first internal transcribed spacer (ITS1) reported previously, sequences of ITS2 and the 5' external transcribed spacer (ETS1) exhibited considerable divergence among these species, including large insertions-deletions detectable by PCR-based spacer length analysis. In slot blot hybridization assays on RNA extracted from lysates of Pseudo-nitzschia cells, oligonucleotide probes directed to pre-rRNA spacers generated much stronger signals than did complementary probes directed to the coding strands of the rDNAs, indicating that the pre-rRNA-targeted probes detected multicopy transcripts. A group of probes directed to a discrete 90-base region within the ITS1 pre-rRNA gave no detectable signal, suggesting that this region is degraded early in the rRNA maturation pathway. Other pre-rRNA regions were always detectable and, in marked contrast to prokaryotic systems analyzed in this manner, were stable and abundant in both actively dividing and nondividing cells. Long, multilabeled RNA probes, which would exhibit considerable cross-reactivity if directed to mature rRNA sequences, detected species-specific pre-rRNA sequences from as few as 1,000 cells. Pre-rRNA is a potentially useful molecular target for detecting and identifying Pseudo-nitzschia species and possibly other unicellular eukaryotes as well. PMID- 9406404 TI - Biochemical and molecular characterization of the polyhydroxybutyrate depolymerase of Comamonas acidovorans YM1609, isolated from freshwater. AB - Comamonas acidovorans YM1609 secreted a polyhydroxybutyrate (PHB) depolymerase into the culture supernatant when it was cultivated on poly(3-hydroxybutyrate) [P(3HB)] or poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV)] as the sole carbon source. The PHB depolymerase was purified from culture supernatant of C. acidovorans by two chromatographic methods, and its molecular mass was determined as 45,000 Da by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The enzyme was stable at temperatures below 37 degrees C and at pH values of 6 to 10, and its activity was inhibited by diisopropyl fluorophosphonate. The liquid chromatography analysis of water-soluble products revealed that the primary product of enzymatic hydrolysis of P(3HB) was a dimer of 3-hydroxybutyric acid. Kinetics of enzymatic hydrolysis of P(3HB) film were studied. In addition, a gene encoding the PHB depolymerase was cloned from the C. acidovorans genomic library. The nucleotide sequence of this gene was found to encode a protein of 494 amino acids (M(r), 51,018 Da). Furthermore, by analysis of the N-terminal amino acid sequence of the purified enzyme, the molecular mass of the mature enzyme was calculated to be 48,628 Da. Analysis of the deduced amino acid sequence suggested a domain structure of the protein containing a catalytic domain, fibronectin type III module as linker, and a putative substrate binding domain. Electron microscopic visualization of the mixture of P(3HB) single crystals and a fusion protein of putative substrate-binding domain with glutathione S-transferase demonstrated that the fusion protein adsorbed strongly and homogeneously to the surfaces of P(3HB) single crystals. PMID- 9406407 TI - Purification and partial characterization of a tripeptidase from Pediococcus pentosaceus K9.2. AB - A tripeptidase was purified from the cytoplasm of Pediococcus pentosaceus K9.2 by anion-exchange chromatography, gel filtration chromatography, and high performance liquid chromatography. The molecular mass of the enzyme was estimated by gel filtration at 100,000 Da. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified peptidase showed one protein band of 45,000 Da. Optimal enzyme activity was obtained at pH 7.0 and at 50 degrees C. The peptidase hydrolyzed all tripeptides tested. Cleavage was not observed with dipeptides, oligopeptides, or amino acid-p-nitroanilide derivatives. Strong inhibition of activity was caused by EDTA, 1,10-phenanthroline, dithiothreitol, and beta mercaptoethanol, whereas phenylmethylsulfonyl fluoride and sulfur-reactive reagents had no effect on peptidase activity. Mg2+, Mn2+, and Ca2+ stimulated the hydrolyzing activity of the enzyme. The 20 N-terminal amino acids of the tripeptidase from P. pentosaceus had 84% identity with those from the corresponding N-terminal region of the tripeptidase from Lactococcus lactis subsp. cremoris Wg2. PMID- 9406406 TI - Construction of a Helicobacter pylori-Escherichia coli shuttle vector for gene transfer in Helicobacter pylori. AB - In this study, a Helicobacter pylori-Escherichia coli shuttle vector was constructed for transferring DNA into H. pylori. The smallest cryptic plasmid (1.2 kb), pHP489, among those harbored by 77 H. pylori isolates was selected as a base replicon for constructing vectors. HindIII-digested pHP489 was ligated with a kanamycin resistance gene [aph(3')-III], which originated from Campylobacter jejuni, to produce the recombinant plasmid pHP489K. pHP489K was efficiently transformed into and stably maintained in H. pylori strains. The shuttle vector pBHP489K (3.6 kb) was constructed by the recombination of pHP489, ColE1, and aph(3')-III sequences. pBHP489K was reciprocally transformed into and maintained in both H. pylori and E. coli. Introduction of the shuttle vector clone DNA (pBHP489K/AB; 6.7 kb), containing the ureA and ureB genes of H. pylori, into urease-negative mutants of H. pylori led to the restoration of their urease activity. The transformants were confirmed to contain the incoming plasmid DNA. pBHP489K satisfied the requirements for an H. pylori-E. coli shuttle vector, implying that it might be a useful vector for investigating pathogenicity and restriction-modification systems of H. pylori. PMID- 9406408 TI - A chloride-inducible gene expression cassette and its use in induced lysis of Lactococcus lactis. AB - A chloride-inducible promoter previously isolated from the chromosome of Lactococcus lactis (J. W. Sanders, G. Venema, J. Kok, and K. Leenhouts, Mol. Gen. Genet., in press) was exploited for the inducible expression of homologous and heterologous genes. An expression cassette consisting of the positive-regulator gene gadR, the chloride-inducible promoter Pgad, and the translation initiation signals of gadC was amplified by PCR. The cassette was cloned upstream of Escherichia coli lacZ, the holin-lysin cassette (lytPR) of the lactococcal bacteriophage r1t, and the autolysin gene of L. lactis, acmA. Basal activity of Pgad resulted in a low level of expression of all three proteins. Growth in the presence of 0.5 M NaCl of a strain containing the gadC::lacZ fusion resulted in a 1,500-fold increase of beta-galactosidase activity. The background activity levels of LytPR and AcmA had no deleterious effects on cell growth, but induction of lysin expression by addition of 0.5 M NaCl resulted in inhibition of growth. Lysis was monitored by following the release of the cytoplasmic marker enzyme PepX. Released PepX activity was maximal at 1 day after induction of lytPR expression with 0.1 M NaCl. Induction of acmA expression resulted in slower release of PepX from the cells. The presence of the inducing agent NaCl resulted in the stabilization of osmotically fragile cells. PMID- 9406409 TI - Extended screening by PCR for seven cry-group genes from field-collected strains of Bacillus thuringiensis. AB - An extended multiplex PCR method was established to rapidly identify and classify Bacillus thuringiensis strains containing cry (crystal protein) genes toxic to species of Lepidoptera, Coleoptera, and Diptera. The technique enriches current strategies and simplifies the initial stages of large-scale screening of cry genes by pinpointing isolates that contain specific genes or unique combinations of interest with potential insecticidal activities, thus facilitating subsequent toxicity assays. Five pairs of universal primers were designed to probe the highly conserved sequences and classify most (34 of about 60) genes known in the following groups: 20 cry1, 3 cry2, 4 cry3, 2 cry4, 2 cry7, and 3 cry8 genes. The DNA of each positive strain was probed with a set of specific primers designed for 20 of these genes and for cry11A. Twenty-two distinct cry-type profiles were identified from 126 field-collected B. thuringiensis strains. Several of them were found to be different from all published profiles. Some of the field collected strains, but none of the 16 standard strains, were positive for cry2Ac. Three standard and 38 field-collected strains were positive by universal primers but negative by specific primers for all five known genes of cry7 and cry8. These field-collected strains seem to contain a new gene or genes that seem promising for biological control of insects and management of resistance. PMID- 9406410 TI - Degradation of tetrahydrofurfuryl alcohol by Ralstonia eutropha is initiated by an inducible pyrroloquinoline quinone-dependent alcohol dehydrogenase. AB - An organism tentatively identified as Ralstonia eutropha was isolated from enrichment cultures containing tetrahydrofurfuryl alcohol (THFA) as the sole source of carbon and energy. The strain was able to tolerate up to 200 mM THFA in mineral salt medium. The degradation was initiated by an inducible ferricyanide dependent alcohol dehydrogenase (ADH) which was detected in the soluble fraction of cell extracts. The enzyme catalyzed the oxidation of THFA to the corresponding tetrahydrofuran-2-carboxylic acid. Studies with n-pentanol as the substrate revealed that the corresponding aldehyde was released as a free intermediate. The enzyme was purified 211-fold to apparent homogeneity and could be identified as a quinohemoprotein containing one pyrroloquinoline quinone and one covalently bound heme c per monomer. It was a monomer of 73 kDa and had an isoelectric point of 9.1. A broad substrate spectrum was obtained for the enzyme, which converted different primary alcohols, starting from C2 compounds, secondary alcohols, diols, polyethylene glycol 6000, and aldehydes, including formaldehyde. A sequence identity of 65% with a quinohemoprotein ADH from Comamonas testosteroni was found by comparing 36 N-terminal amino acids. The ferricyanide-dependent ADH activity was induced during growth on different alcohols except ethanol. In addition to this activity, an NAD-dependent ADH was present depending on the alcohol used as the carbon source. PMID- 9406411 TI - Acquisition of a deliberately introduced phenol degradation operon, pheBA, by different indigenous Pseudomonas species. AB - Horizontal transfer of genes of selective value in an environment 6 years after their introduction into a watershed has been observed. Expression of the gene pheA, which encodes phenol monooxygenase and is linked to the pheBA operon (A. Nurk, L. Kasak, and M. Kivisaar, Gene 102:13-18, 1991), allows pseudomonads to use phenol as a growth substrate. Pseudomonas putida strains carrying this operon on a plasmid were used for bioremediation after an accidental fire in the Estonia oil shale mine in Estonia in 1988. The water samples used for studying the fate of the genes introduced were collected in 1994. The same gene cluster was also detected in Pseudomonas strains isolated from water samples of a nearby watershed which has been continuously polluted with phenols due to oil shale industry leachate. Together with the more frequently existing counterparts of the dmp genes (V. Shingler, J. Powlowski, and U. Marklund, J. Bacteriol. 174:711-724, 1992), the pheA gene was also represented in the phenol-degrading strains. The area where the strains containing the pheA gene were found was restricted to the regular route of phenolic leachate to the Baltic Sea. Nine Pseudomonas strains belonging to four different species (P. corrugata, P. fragi, P. stutzeri, and P. fluorescens biotypes B, C, and F) and harboring horizontally transferred pheBA operons were investigated. The phe genes were clustered in the same manner in these nine phe operons and were connected to the same promoter as in the case of the original pheBA operon. One 10.6-kb plasmid carrying a pheBA gene cluster was sequenced, and the structure of the rearranged pheBA operon was described. This data indicates that introduced genetic material could, if it encodes a beneficial capability, enrich the natural genetic variety for biodegradation. PMID- 9406412 TI - Distribution of metabolic activity and phosphate starvation response of lux tagged Pseudomonas fluorescens reporter bacteria in the barley rhizosphere. AB - The purpose of this study was to determine the metabolic activity of Pseudomonas fluorescens DF57 in the barley rhizosphere and to assess whether sufficient phosphate was available to the bacterium. Hence, two DF57 reporter strains carrying chromosomal luxAB gene fusions were introduced into the rhizosphere. Strain DF57-40E7 expressed luxAB constitutively, making bioluminescence dependent upon the metabolic activity of the cells under defined assay conditions. The DF57 P2 reporter strain responded to phosphate limitation, and the luxAB gene fusion was controlled by a promoter containing regulatory sequences characteristic of members of the phosphate (Pho) regulon. DF57 generally had higher metabolic activity in a gnotobiotic rhizosphere than in the corresponding bulk soil. Within the rhizosphere the distribution of metabolic activity along the root differed between the rhizosphere soil and the rhizoplane, suggesting that growth conditions may differ between these two habitats. The DF57-P2 reporter strain encountered phosphate limitation in a gnotobiotic rhizosphere but not in a natural rhizosphere. This difference in phosphate availability seemed to be due to the indigenous microbial population, as DF57-P2 did not report phosphate limitation when established in the rhizosphere of plants in sterilized soil amended with indigenous microorganisms. PMID- 9406414 TI - Purification, characterization, and nucleotide sequence of an intracellular maltotriose-producing alpha-amylase from Streptococcus bovis 148. AB - An intracellular alpha-amylase from Streptococcus bovis 148 was purified and characterized. The enzyme was induced by maltose and soluble starch and produced about 80% maltotriose from soluble starch. Maltopentaose was hydrolyzed to maltotriose and maltose and maltohexaose was hydrolyzed mainly to maltotriose by the enzyme. Maltotetraose, maltotriose, and maltose were not hydrolyzed. This intracellular enzyme was considered to be a maltotriose-producing enzyme. The enzymatic characteristics and hydrolysis product from soluble starch were different from those of the extracellular raw-starch-hydrolyzing alpha-amylase of strain 148. The deduced amino acid sequence of the intracellular alpha-amylase was similar to the sequences of the mature forms of extracellular liquefying alpha-amylases from Bacillus strains, although the intracellular alpha-amylase did not contain a signal peptide. No homology between the intracellular and extracellular alpha-amylases of S. bovis 148 was observed. PMID- 9406415 TI - A mutant of Listeria monocytogenes LO28 unable to induce an acid tolerance response displays diminished virulence in a murine model. AB - Exposing Listeria monocytogenes LO28 to sublethal pH induces protection against normally lethal pH conditions, a phenomenon known as the acid tolerance response. We identified a mutant, L. monocytogenes ATR1, which is incapable of inducing such tolerance, either against low pH or against any other stress tested. The virulence of this mutant was considerably decreased, suggesting that the acid tolerance response contributes to in vivo survival of L. monocytogenes. PMID- 9406416 TI - Direct detection and isolation of plasmid-bearing virulent serotypes of Yersinia enterocolitica from various foods. AB - A procedure was developed for direct detection, isolation, and maintenance of plasmid-bearing virulent serotypes of Yersinia enterocolitica from different food sources. Plasmid-bearing virulent strains of Y. enterocolitica representing five serotypes were simultaneously detected and isolated from enriched swab samples of artificially contaminated pork chops, ground pork, cheese, and zucchini, using Congo red binding and low-calcium-response tests. The method was also effective in isolating plasmid-bearing virulent strains of Y. enterocolitica from naturally contaminated porcine tongues. Virulence of the strains isolated from these foods was confirmed by PCR, the expression of plasmid-associated phenotypes, and mouse pathogenicity. PMID- 9406417 TI - Recombinant thermostable cycloinulo-oligosaccharide fructanotransferase produced by Saccharomyces cerevisiae. AB - A truncated fragment of the cycloinulo-oligosaccharide fructanotransferase (CFTase) gene of Bacillus circulans MCI-2554 was fused to the prepro secretion sequence of the alpha-factor and expressed in Saccharomyces cerevisiae under the control of the 5' upstream region of the isocitrate lyase gene of Candida tropicalis (UPR-ICL). Efficiently secreted recombinant CFTase protein (yeast CFTase) was purified. Yeast CFTase consisted of three protein molecules, each of which had CFTase activity (yeast CFTase 1 [116 kDa], yeast CFTase 2 [117 kDa], and yeast CFTase 3 [116 kDa]). Yeast CFTase 2 was the major product of the expression system employed and was shown to be N glycosylated by endoglycosidase H treatment. Yeast CFTase 1 was N glycosylated but had a short truncation at its N terminus, while yeast CFTase 3 did not contain an N-glycosylated carbohydrate chain(s). Yeast CFTase 2 showed an optimum pH, an optimum temperature, and a pH stability similar to those of CFTase purified from B. circulans but exhibited a significant increase in thermostability. Production of yeast CFTase by the strain which had two copies of the CFTase gene integrated into its chromosomes reached 391 U per liter of culture at 120 h, which corresponded to 8.40 mg of protein per liter, by shake-flask cultivation. PMID- 9406418 TI - Conditions for natural transformation of Ralstonia solanacearum. AB - The development of competence allowing natural transformation of Ralstonia solanacearum was found to occur during exponential growth and not in response to any excreted factors. Linear DNAs were effectively integrated by recombination requiring a minimum of 50 bp of homologous DNA. Therefore, DNA from other genera and species were ineffective. PMID- 9406419 TI - Inactivation of Helicobacter pylori by chlorination. AB - Three strains of Helicobacter pylori were studied to determine their resistance to chlorination. The organisms were readily inactivated by free chlorine and should therefore be controlled by disinfection practices normally employed in the treatment of drinking water. PMID- 9406420 TI - Mosquito larvicidal activity of transgenic Anabaena strain PCC 7120 expressing combinations of genes from Bacillus thuringiensis subsp. israelensis. AB - Various combinations of the genes cryIVA (cry4A), cryIVD (cry11A), and p20 from Bacillus thuringiensis subsp. israelensis were introduced into the nitrogen fixing cyanobacterium Anabaena sp. strain PCC 7120 by means of Escherichia coli Anabaena shuttle vector pRL488p and were expressed under control of two tandem strong promoters, a cyanobacterial promoter (PpsbA) and an E. coli T7 promoter (PA1). Two of the clones carrying cryIVA plus cryIVD, one with p20 and one without p20, displayed toxicity against third-instar larvae of Aedes aegypti at levels greater than any level previously reported for transgenic cyanobacteria. PMID- 9406421 TI - Heat inactivation of Mycobacterium paratuberculosis in raw milk: are current pasteurization conditions effective? AB - Currently, it is not known whether commercial pasteurization effectively kills Mycobacterium paratuberculosis in contaminated raw milk. Results from holder test tube experiments indicated that a residual population of viable bacteria remained after treatment at 65, 72, 74, or 76 degrees C for 0 to 30 min. Use of a laboratory-scale pasteurizer unit demonstrated that treatment of raw milk at 72 degrees C for 15 s effectively killed all M. paratuberculosis. PMID- 9406423 TI - Vibrio sp. strain NM 10, isolated from the intestine of a Japanese coastal fish, has an inhibitory effect against Pasteurella piscicida. AB - Vibrio sp. strain NM 10 with an inhibitory activity against Pasteurella piscicida K-III was isolated from the intestine of a spotnape ponyfish (Leiognathus nuchalis). This bacterium efficiently produced an antibacterial substance after growth at 20 degrees C for 24 h on 1/5 PYBG agar prepared with 50% seawater at pHs of 7.5 to 9.0. The antibacterial substance was heat labile and proteinaceous, with a molecular mass of less than 5 kDa, possibly a bacteriocin or a bacteriocin like substance. PMID- 9406424 TI - One-step purification of nisin A by immunoaffinity chromatography. AB - The lantibiotic nisin A was purified to homogeneity by a single-step immunoaffinity chromatography method. An immunoadsorption matrix was developed by direct binding of anti-nisin A monoclonal antibodies to N-hydroxysuccinimide activated Sepharose. The purification procedure was rapid and reproducible and rendered much higher final yields of nisin than any other described method. PMID- 9406426 TI - Adaptation of microorganisms and their transport systems to high temperatures. AB - Growth of Bacteria and Archaea has been observed at temperatures up to 95 and 110 degrees C, respectively. These thermophiles are adapted to environments of high temperature by changes in the membrane lipid composition, higher thermostabilities of the (membrane) proteins, higher turnover rates of the energy transducing enzymes, and/or the (exclusive) use of sodium-ions rather than protons as coupling ion in energy transduction. The proton permeability of the cytoplasmic membrane of bacteria and archaea was observed to increase with the temperature. This increased proton permeability limits the maximum temperature of growth of bacteria. Higher growth temperatures can be reached by an increased proton pumping activity by using the less permeable sodium ions as coupling ions or by changing the lipid composition of the cytoplasmic membrane. The Na+/H+/glutamate transport proteins of the thermophiles Bacillus stearothermophilus (GltTBs) and Bacillus caldotenax (GltTBc) were studied extensively. These transportproteins have unique features. Transport of L glutamate occurs in symport with 1 Na+ and 1 H+ when the transport proteins are expressed in their natural environment. The sodium ion dependency of the GltT transporters of these Bacillus strains was found to increase with temperature. However, when the GltT proteins are expressed in the mesophile Escherichia coli, electrogenic symport of L-glutamate occurs with > or = 2 H+. These observations suggest that the conformation of the transport proteins in the E. coli and the Bacillus membranes differs, and that the conformation influences the coupling ion selectivity. The Na+/H+/glutamate transport proteins of B. stearothermophilus (GltTBs) and B. caldotenax (GltTBc) are homologous to transport systems of glutamate and structurally related compounds from mesophilic organisms. Both sodium, as well as proton coupled transporters, belong to this family of carboxylate transporters (FCT). PMID- 9406427 TI - Thermophilic proteins: stability and function in aqueous and organic solvents. AB - The molecular stability of thermophilic and hyperthermophilic enzymes generally reflects the growth temperatures of the parent organisms. Extracellular enzymes from the hyperthermophilic Archaea typically show very high levels of thermal stability and a number of enzymes with Tm values of greater than 100 degrees C have been reported. The mechanisms responsible for high molecular stability are typically intrinsic characteristics of the protein, as shown by the comparative stabilities of many native and recombinant proteins. However, some extrinsic stabilisation mechanisms have been demonstrated. High levels of thermal stability are positively correlated with stability in the presence of other denaturing agents, including detergents and organic solvents. This correlation suggests a common denaturation pathway where molecular mobility/flexibility is the prime determinant of susceptibility to irreversible denaturation. In single phase organic-aqueous solvents, protein destabilisation occurs via solvent-induced alteration to the protein hydration shell. However, correlations between protein stability and solvent hydrophobicity are unreliable. In two-phase organic-aqueous systems, interfacial denaturation predominates and is a function of both interfacial tension and interfacial surface area. Intracellular enzymes are protected from interfacial denaturation but are potentially susceptible to direct organic solvent effects, possibly depending on the role of the cell wall and cell membrane in the partitioning of the organic solvent into the cell cytoplasm. Immobilisation of thermophilic enzymes provides a method for enhancing both the thermal and solvent stabilities of thermophilic and mesophilic enzymes. Multi point covalent immobilisation to glyoxal-agarose enhances thermal stability and limits protein-protein inactivation mechanisms. Miscible organic solvents have a profound influence on the specificities of enzyme reactions. The presence of high concentrations of miscible organic solvents may induce gross changes in substrate specificity and/or more subtle alterations in chiral selectivity. Correlations between the variation in enantioselectivity and both solvent hydrophobicity and solvent dielectric constant have been demonstrated although some recent studies implicate the formation of specific solvent-enzyme complexes which directly affect reaction kinetics. PMID- 9406428 TI - Sequence and structural comparison of thermophilic phosphoglycerate kinases with a mesophilic equivalent. AB - The monomeric glycolytic enzyme phosphoglycerate kinase (PGK) has been used as a model system to study protein thermostability. The primary sequence of this enzyme has been elucidated from 47 species to date. Although only 42 amino acids are totally conserved, most of which line the active site cleft, the protein is structurally conserved. This is achieved by making conservative changes to maintain the same secondary and tertiary folds. The crystal structures of 5 PGK enzymes have been solved by X-ray diffraction methods. This paper seeks to use the available information to understand protein thermostability. Although some general mechanisms to increase stability can be determined, different species have adopted a variety of subtle additive changes to achieve greater protein stability. Comparisons have been directly made between the PGK enzyme from yeast, the moderate thermophilic bacterium Bacillus stearothermophilus, the hyperthermophilic bacteria Thermus thermophilus, Thermotoga maritima, and the hyperthermophilic archaea Sulpholobus solfataricus and Methanothermus fervidus. PMID- 9406429 TI - Molecular physiology of carbamoylation under extreme conditions: what can we learn from extreme thermophilic microorganisms? AB - The importance of protein-protein interactions in the physiology of extreme thermophiles was investigated by analyzing the enzymes involved in biosynthetic carbamoylation in Thermus ZO5 and by comparing the results obtained with already available or as yet unpublished information concerning other thermophilic eu- and archaebacteria such as Thermotoga, Sulfolobus, and Pyrococcus. Salient observations were that (i) the highly thermolabile and reactive carbamoylphosphate molecule appears to be protected from thermodegradation by channelling towards the synthesis of citrulline and carbamoylaspartate, respectively precursors of arginine and the pyrimidines; (ii) Thermus ornithine carbamoyltransferase is clearly a thermophilic enzyme, intrinsically thermostable and showing a biphasic Arrhenius plot, whereas aspartate carbamoyltransferase is inherently unstable and is stabilized by its association with dihydroorotase, another enzyme encoded by the Thermus pyrimidine operon. Possible implications of these results are discussed. PMID- 9406430 TI - Acidostable and acidophilic proteins: the example of the alpha-amylase from Alicyclobacillus acidocaldarius. AB - Acidophilic microorganisms grow optimally at pH values between 1-4. They have adapted to the acid condition by maintaining their cytoplasmic pH at a value close to neutrality. Hence, only those (macro)-molecules, which face the acid medium, have had to adapt to this extreme condition. Literature data show that several exoproteins from thermoacidophilic prokaryotes are characterized by a low charge density. It is proposed that this property contributes to the stability of these proteins both below and above the pKa-values of their glutamate and aspartate residues. As an example of an acidophilic protein, the alpha-amylase from the Gram-positive Alicyclobacillus acidocaldarius ATCC27009 was studied. The enzyme is thermoacidophilic, with optima of temperature and pH of 75 degrees C and pH 3, respectively. The nucleotide sequence of the cloned gene (8) indicates that the alpha-amylase belongs to a large family of starch-degrading enzymes with a characteristic catalytic (beta alpha)8-domain. Three essential and probably catalytic acidic residues have been conserved, suggesting that the acidophilic alpha-amylase degrades starch with essentially the same mechanism as do its neutrophilic relatives. Still, the acidophilic protein contains three exchanges in residues uniformally or almost uniformally conserved among all members of the enzyme family. In order to test whether these exchanges contribute to the acidic pH optimum, the alpha-amylase gene was expressed in Escherichia coli. Sonication of the enzyme-producing cells released alpha-amylase activity associated with a 140 kDa protein. The optima of temperature and pH for the protein produced in E. coli were similar to those of the native enzyme. Experiments are underway in which it is tested which residues contribute to the acid pH optimum of the alpha amylase. PMID- 9406431 TI - Psychrophilic bacteria--molecular adaptations of membrane lipids. PMID- 9406432 TI - Microtubule assembly in cold-adapted organisms: functional properties and structural adaptations of tubulins from antarctic fishes. AB - Fishes native to the coastal waters of the Antarctic have adapted to habitat and body temperatures in the range -1.8 to +2 degrees C. Their cytoplasmic microtubules, unlike those of mammals and temperate poikilotherms, have evolved to assemble efficiently at these low temperatures. To learn about the underlying molecular adaptations, my laboratory is studying microtubule proteins [tubulin alpha beta dimers and microtubule-associated proteins (MAPs)] and tubulin genes from several Antarctic fishes, including the rockcods Notothenia coriiceps and Gobionotothen gibberifrons. We find that the assembly-enhancing adaptations of the fish microtubule proteins are intrinsic to the tubulin subunits themselves. Furthermore, microtubule formation by Antarctic fish tubulins is strongly entropy driven, due in part to an increased reliance, relative to tubulins from other species, on hydrophobic interactions. Based on analyses of tubulin polypeptides and cDNAs, we suggest that the structural adaptations of Antarctic fish tubulins most likely involve alterations in the primary sequences of tubulin isotypes. With respect to neural beta tubulins from other vertebrates, for example, the class II beta-tubulin isotype of N. coriiceps brain contains seven unique amino acid substitutions and one novel insertion in its 446-residue primary sequence. Most of these changes are located in a structural domain that forms contacts between tubulin dimers during microtubule assembly and would be expected to enhance polypeptide flexibility, thereby facilitating addition of tubulin to microtubule ends. The acidic carboxy-terminal tails of the alpha and beta tubulins, by contrast, appear not to be sites of cold adaptation of polymerization. We have also found that brain and egg tubulins from Antarctic fishes differ strikingly in their polymerization efficiencies, which demonstrates, in agreement with the multitubulin hypothesis, that tissue-specific tubulin isoforms can possess distinct functional properties. Thus, study of microtubule proteins from organisms, such as the Antarctic fishes, that have adapted to extreme thermal regimes should contribute significantly to an understanding of the quaternary interactions that control microtubule assembly in all eukaryotes. PMID- 9406433 TI - Background adaptation by Xenopus laevis: a model for studying neuronal information processing in the pituitary pars intermedia. AB - This review is concerned with recent literature on the neural control of the pituitary pars intermedia of the amphibian Xenopus laevis. This aquatic toad adapts skin colour to the light intensity of its environment, by releasing the proopiomelanocortin (POMC)-derived peptide alpha-MSH (alpha-melanophore stimulating hormone) from melanotrope cells. The activity of these cells is controlled by brain centers of which the hypothalamic suprachiasmatic and magnocellular nuclei, respectively, inhibit and stimulate both biosynthesis and release of alpha-MSH. The suprachiasmatic nucleus secretes dopamine, GABA, and NPY from synaptic terminals on the melanotropes. The structure of the synapses depends on the adaptation state of the animal. The inhibitory transmitters act via cAMP. Under inhibition conditions, melanotropes actively export cAMP, which might have a first messenger action. The magnocellular nucleus produces CRH and TRH. CRH, acting via cAMP, and TRH stimulate POMC-biosynthesis and POMC-peptide release. ACh is produced by the melanotrope cell and acts in an autoexcitatory feedback on melanotrope M1 muscarinic receptors to activate secretory activity. POMC-peptide secretion is driven by oscillations of the [Ca2+]i, which are initiated by receptor-mediated stimulation of Ca2+ influx via N-type calcium channels. The hypothalamic neurotransmitters and ACh control Ca2+ oscillatory activity. The structural and functional aspects of the various neural and endocrine steps in the regulation of skin colour adaptation by Xenopus reveal a high degree of plasticity, enabling the animal to respond optimally to the external demands for physiological adaptation. PMID- 9406434 TI - Nongenetic variation, genetic-environmental interactions and altered gene expression. III. Posttranslational modifications. AB - The use of protein electrophoretic data for determining the relationships among species or populations is widespread and generally accepted. However, posttranslational modifications have been discovered in many of the commonly analyzed proteins and enzymes. Posttranslational modifications often alter the electrophoretic mobility of the modified enzyme or protein. Because posttranslational modifications may affect only a fraction of the total enzyme or protein, an additional staining band often appears on gels as a result, and this may confound interpretations. Deamidation, acteylation, proteolytic modification, and oxidation of sulfhydryl groups are modifications that often result in an electrophoretic mobility shift. Sialic acid-induced heterogeneity has been documented for many enzymes, but neuraminidase treatment can often remove sialic acids and produce gel patterns that are easier to interpret. In some cases, ontogenetic and tissue-specific expression may be due to posttranslational modifications rather than gene control and restricted expression, respectively. Methods of preventing, detecting and eliminating posttranslational modifications are discussed. Some posttranslational modifications may be useful for detecting cryptic genetic polymorphisms. PMID- 9406435 TI - Is the flow in the giraffe's jugular vein a "free" fall? AB - There is controversy as to whether or not the heart works against gravity in the arteries to the head in the upright position. One view is that the gravitational effects in the neck arteries are counterbalanced by the gravitational effects in the veins of the neck and the heart does not do extra pressure work. This concept has been challenged by others who claim that the heart works against gravity based on the notion that the jugular vein is collapsed and gravitational effects on jugular blood are inoperative, similar to the "free" fall of liquids. The present study supports the view that blood flow in the collapsible jugular vein of the giraffe is not a "free" fall and that the heart does not spend extra energy to raise the blood to the head. PMID- 9406436 TI - Ouabain-sensitive Na+,K(+)-ATPase activity in toad brain. AB - Toads of the genus Bufo are highly resistant to the toxic effects of digitalis glycosides, and the Na+,K(+)-ATPase of all toad tissues studied to date has been relatively insensitive to inhibition by digitalis and related compounds. In studies of brain microsomal preparations from two toad species, Bufo marinus and Bufo viridis, inhibition of ATPase activity and displacement of [3H]ouabain from Na+,K(+)-ATPase occurred over broad ranges of ouabain or bufalin concentrations, consistent with the possibility that more than one Na+,K(+)-ATPase isoform may be present in toad brain. The data could be fitted to one- or two-site models, both of which were consistent with the presence of Na+,K(+)-ATPase activity with high sensitivity to ouabain and bufalin. Ki (concentration capable of producing 50% inhibition of activity) values for ouabain in the one-site model were in the 0.2 to 3.7 microM range, whereas Ki1 values in the two-site model ranged from 0.085 to 0.85 microM, indicating that brain ATPase was at least three orders of magnitude more sensitive to ouabain than B. marinus bladder ATPase (Ki = 5940 microM). Ouabain was also an effective inhibitor of 86Rb+ uptake in B. marinus brain tissue slices (Ki = 3.1 microM in the one-site model; Ki1 = 0.03 microM in the two-site model). However, the relative contribution of the high ouabain sensitivity site to the total activity was 17% in the transport assay as compared with 63% in the Na+,K(+)-ATPase enzymatic assay. We conclude that a highly ouabain-sensitive Na+,K(+)-ATPase activity is present and functional in toad brain but that its function may be partially inhibited in vivo. PMID- 9406437 TI - Measurement of elastic properties of Xenopus oocytes. AB - Elastic properties of Xenopus oocytes were examined by measuring intracellular pressure (Pic) and cell volume (Vc) in cells undergoing osmotic swelling. Pic was measured by micropuncture, using the servo-null technique. Vc was obtained by analyzing images acquired from a microscope having a video camera attachment. During osmotic swelling, Pic increased from 61 +/- 17 to 500 +/- 59 Pa (mean +/- SE), but the relationship with volume was not linear. In cells that underwent sequential swelling and shrinking, Pic was always lower on shrinking and the cells showed hysteresis. Cells with vitelline envelope (VE) removed had Pic-Vc curves similar in shape to those of intact cells; however, Pic values were significantly lower. Specific elastance[delta Pic/(delta Vc/Vc)] was reduced by removal of the VE. The data indicate that oocytes are weakly elastic and that a large part of their resistance to expansion resides in the VE. PMID- 9406439 TI - Feeding and digesting fiber and tannins by an herbivorous rodent, Octodon degus (Rodentia:Caviomorpha). AB - Differences in feeding rates and digestive efficiency of alternative experimental diets differing in cellulose or fiber and a secondary metabolite (the hydrolyzable tannin, tannic acid [TA]) were assessed with the herbivorous burrowing caviomorph rodent Octodon degus (degu). Degus live in open scrub subjected to summer droughts. The in vitro activity of the digestive enzyme sucrase was not significantly different between treatments with high and low TA. Analysis of the whole organism allowed us to conclude that in vitro analyses of enzymatic digestive activity and plant defenses cannot be used to explain and fully understand the physiological and behavioral effects of plant defenses on mammalian herbivores. We observed no body mass reduction due to effects of dietary treatments. O. degus seemed to compensate for nutritionally poor food by increasing gut content volume. We conclude that fiber and secondary compounds may influence feeding and digestive strategies and vice versa. PMID- 9406438 TI - Levels of biogenic amines in larvae and adults of the rat hookworm, Nippostrongylus brasiliensis (Nematoda). AB - Norepinephrine, 5-hydroxytryptophan, octopamine, dihydroxyphenylacetic acid, N acetyldopamine, dopamine, 5-hydroxyindoleacetic acid, N-acetylserotonin, tyramine, tryptophan and serotonin in larvae (third free stage and parasitic stages) and adult males and females (at defined ages during the intestinal phase) of the parasitic nematode Nippostrongylus brasiliensis were quantified simultaneously by high-performance liquid chromatography with electrochemical detection. Biogenic amine levels depended on the stage, the age and the sex of parasites and on environmental conditions. Their physiological roles in reproductively competent adults of this nematode are discussed in relation to exuviation and egg laying. Parallel fluctuations in free ecdysteroids and norepinephrine were observed in females from the same worm populations. PMID- 9406441 TI - Stress affects corticosteroid and immunoglobulin concentrations in male house mice (Mus musculus) and prairie voles (Microtus ochrogaster). AB - Glucocorticoids, secreted in response to perceived stress, can suppress immunoglobulin (Ig) levels and compromise immune function in mice and rats. Prairie voles (Microtus ochrogaster) have been reported to exhibit basal corticosterone concentrations that would cause pathological changes in the immune function of most other rodents. The goals of the present study were to verify that serum corticosterone concentrations are high in prairie voles, as compared with house mice (Mus musculus), by measuring serum corticosterone with the same RIA; to examine the effects of mild stressors on corticosterone response in both species and to examine the effects of elevated corticosterone levels on IgM and IgG levels in prairie voles and house mice. After 2 weeks of randomly timed 15 min daily restraint or cold-water swim sessions, animals were injected with sheep red blood cells. The data confirmed that basal blood concentrations of corticosterone were higher in prairie voles than house mice, but these high levels doubled after the first swim session in prairie voles, indicating that the adrenals can respond to stressors by producing increased corticosterone. After stress, antibody production (both IgM and IgG) was reduced in house mice but not in prairie voles, despite higher blood concentrations of glucocorticoids in prairie voles. Although body mass was statistically equivalent between species, prairie voles and mice differed dramatically in adrenal and splenic masses. Average adrenal mass of prairie voles was approximately three times the average mass of these organs in house mice; in contrast, the average splenic mass of house mice was approximately three times that of prairie voles. These data may be relevant to seasonal changes in immune function and survival. PMID- 9406440 TI - L-alanine uptake by frog (Rana esculenta) red blood cells. AB - L-Alanine uptake has been studied in frog red blood cells. The present study shows the presence of different carriers for this amino acid in these cells. In the physiological concentration range, most L-alanine is taken up through the Na(+)-dependent system ASC, although the sodium-independent systems asc and L are also active. The competitive inhibition data obtained makes difficult to differentiate the two Na(+)-independent activities in a clear contrast with data from fish or mammalian erythrocytes, indicating that despite its widespread occurrence in vertebrates, these carriers show characteristics that are species specific. PMID- 9406443 TI - Evaporative water loss and oxygen uptake in two casque-headed tree frogs, Aparasphenodon brunoi and Corythomantis greeningi (Anura, Hylidae). AB - Evaporative water loss (EWL) and oxygen uptake (Vo2) was measured in two species of tree frogs with cranial co-ossification, Aparasphenodon brunoi and Corythomantis greeningi. Both species use their head to seal the entrance of bromeliads, tree holes or rocky crevices used as shelters. EWL was significantly reduced in sheltered individuals of both species as compared with those exposed nude to desiccation. EWL per unit area through the head surface was significantly lower than the body skin for A. brunoi but not for C. greeningi, EWL per unit surface area through C. greeningi body skin was about 50% that of A. brunoi, indicating a less permeable skin in the former species. The relationship between cranial coossification and EWL is discussed. Vo2 in A. brunoi was comparable with other anurans of similar size, whereas in C. greeningi, it was lower than predicted from body mass. Moreover, Vo2 in C. greeningi showed less sensitivity to temperature increase than in A. brunoi. C. greeningi occurs in a drier environment than A. brunoi, and this appears to be reflected in their EWL and Vo2 characteristics. PMID- 9406442 TI - Gastric Na+K+ATPase activity and intestinal urea hydrolysis of the common vampire bat, Desmodus rotundus. AB - The blood diet of the vampire bat represents an extraordinarily high ratio of protein to other nutrients and the highest water consumption per body weight of any other mammal. This bat has a unique gastrointestinal morphology that is characterized by a reduced small intestine, absence of a large intestine and intestinal cecum and the presence of a water-absorptive gastric fundus. The present study demonstrates that the gastric fundus has a greater Na+K+ATPase activity for active ion transport compared with other equally sized mammals. This activity is believed to be necessary to establish a gradient favoring water absorption across what would otherwise be an osmotic disequilibrium. The absence of a large intestine and intestinal cecum may reflect a reduced urea hydrolysis by the vampire bat. The present study demonstrated that the vampire bat does not hydrolyze urea as does an equally sized non-sanguinivorous mammal. These data suggest that the blood diet and the relocation of water-absorptive tissue from the lower intestinal tract to the stomach is associated with an active ion transport mechanism in the gastric tissue and a reduced capacity for ureolytic microbes to hydrolyze urea in the intestine. Both processes are specializations for a diet high in protein and water. PMID- 9406444 TI - Distribution and reciprocal interactions of 3H-melatonin and 125I-thyroxine in peripheral, neural, and endocrine tissues of bullfrog tadpoles. AB - Tissue distribution of 125I-thyroxine (T4) and 3H-melatonin and the effect of each hormone on the tissue content of the other were studied because previous work indicated that melatonin antagonized metamorphosis through peripheral, as well as thyroidal effects. Late pre- to prometamorphic Rana catesbeiana tadpoles on an 18 light:6 dark cycle were used for injection of hormones in vivo or to supply tissues for in vitro hormone administration. Labeled melatonin uptake was highest in intestine, ventral skin and pituitary; lowest in thyroid and brain and intermediate in hindlimb, tail and gills. The tissue content of labeled T4 was distributed in nearly the same way, except that the thyroid level was relatively higher, and pituitary lower, than that of labeled melatonin. The pineal, studied only in the tracer T4 experiments, had the highest content of labeled T4 of all tissues. Simultaneous injection of either 0.007 or 0.2 microgram T4 increased 3H melatonin uptake into peripheral tissues that undergo major metamorphic changes but not into neural or endocrine organs. In contrast, 0.033, 3.75 or 15 micrograms melatonin had no significant influence on the content of 125I-T4 in any tissue studied in vivo. Results of in vitro labeling of selected tissues were generally in agreement with the in vivo work except that the 125I-T4 content of intestinal segments from late prometamorphic larvae was lower in melatonin treated than in control groups. The results suggest that peripheral tissues are a major site for T4-melatonin interactions and that T4 may modulate its own action through influencing melatonin levels in target tissues and perhaps in the thyroid. Because melatonin had no effect on tissue T4 content in young tadpoles, retardation of metamorphic events by melatonin does not seem to involve modulation of T4 availability to the tissues. PMID- 9406445 TI - Glucose kinetics of the Virginia opossum: possible implications for predicting glucose turnover in mammals. AB - Primed continuous infusions of 6-3H-glucose, respirometry and measurements of nitrogen excretion were carried out in adult Virginia opossums (Didelphis virginiana). Our goals were to determine resting glucose turnover rate, to establish the relative importance of this oxidative fuel in the energy budget of this species, to assess whether metabolic rate is a better predictor of glucose turnover rate than body mass and to demonstrate that glucose kinetics can be measured in the opossum for future use of this animal model in studies of mammalian glucoregulation. Results show that the resting glucose turnover rate of opossums averages 17.5 +/- 0.9 mumol/kg.min (n = 9), that 31% of total glucose flux is oxidized and that glucose oxidation represents 9% of their metabolic rate. An allometric equation predicting glucose turnover rate from body mass for placental mammals overestimates the measured turnover rate of this marsupial by 30%, suggesting that turnover rate is better predicted from metabolic rate than from body mass. Finally, this study demonstrates that the Virginia opossum is a convenient model to study glucose metabolism in vivo, and we propose that the very rapid aging characteristics of this species should be exploited to investigate the effects of aging on glucose homeostasis. PMID- 9406446 TI - Rana esculenta L. liver Fru-1,6-P2ase and G-6-Pase activity and Fru-2,6-P2 concentration after acclimation at 5 and 25 degrees C. AB - The activities of Fru-1,6-P2ase and G-6-Pase in liver and kidney of frogs acclimated at 5 and 25 degrees C and Fru-2,6-P2 level in liver were investigated. The aim of this study was to examine the effect of thermal acclimation on regulatory enzymes of gluconeogenesis and on concentration of gluconeogenesis regulator. Fru-1,6-P2ase activity in liver of frogs acclimated at 5 degrees C was 6.16 +/- 0.77 and 4.46 +/- 0.46 U/g wt in those acclimated at 25 degrees C; the respective values for G-6-Pase were 0.46 +/- 0.04 and 0.25 +/- 0.02 U/g wt. Fru 1,6-P2ase activity in kidney was 3.2 +/- 0.48 U/g wt at 5 degrees C and 2.64 +/- 0.23 U/g wt at 25 degrees C; the respective values for G-6-Pase were 0.2 +/- 0.05 and 0.17 +/- 0.05 U/g wt. K(m) of frog liver Fru-1,6-P2ase determined after acclimation at 5 degrees C and to 25 degrees C was 1.36 and 1.41 microM, respectively. Frog liver Fru-1,6-P2ase was allosterically inhibited by AMP. I0.5 determined after acclimation at 5 degrees C was 10.55 microM and after acclimation at 25 degrees C was 10.88 microM. Liver Fru-2,6-P2 concentration after acclimation at 5 degrees C was 0.44 +/- 0.13 nmol/g wt in comparison with 0.58 +/- 0.19 nmol/g wt after acclimation at 25 degrees C. In conclusion, cold exposure increased hepatic gluconeogenic capacity of Rana esculenta. PMID- 9406448 TI - Exogenous carbohydrate utilisation: effects on metabolism and exercise performance. AB - It is generally recognized that a decrease in carbohydrate availability can lead to the development of fatigue during prolonged exercise in humans. Administration of glucose or other carbohydrates before or during exercise has been shown to postpone fatigue, conserve muscle glycogen and improve performance. Carbohydrates can be categorised according to their ability to increase blood glucose concentration (known as glycaemic index) and by the extent they stimulate the release of insulin. The glycaemic index is reflected in the rate at which consumed carbohydrate is made available in the blood. Glucose is the only type of carbohydrate that can readily be oxidised by skeletal muscle for energy production. Gastric emptying is the primary factor limiting the rate of carbohydrate delivery to the blood and therefore influences the utilisation of exogenous carbohydrate ingested before or during exercise. Various methods have been used to assess the oxidation of exogenous carbohydrates during exercise. Peak rates of CHO oxidation during exercise have been reported between 0.4 and 1.0 g/min, and the rates of oxidation do not appear to be influenced to a major extent by the use of multiple drinking schedule in comparison with a single bolus schedule. Previous studies also suggest that the ingestion of fructose during exercise does not offer any additional benefits over ingestion of glucose or glucose polymer solutions of similar concentration. The hormones insulin, glucagon and adrenaline together with cortisol and growth hormone play key roles in the regulation of carbohydrate metabolism during exercise. Ingestion of moderately concentrated carbohydrate solutions (4-8%) enhances prolonged exercise performance and is appropriate for optimising energy and fluid delivery without causing adverse effects. The ergogenic effects of carbohydrate ingestion on performance during intermittent exercise such as competitive sports are less well established, although the evidence to date suggests diminished performance when carbohydrate are limiting. PMID- 9406447 TI - Effects of proglumide on cholecystokinin-8-induced exocrine and endocrine pancreatic responses in conscious sheep. AB - The effects of cholecystokinin (CCK)-8, either alone or together with the CCK antagonist, proglumide on both exocrine and endocrine pancreatic responses were examined in conscious sheep. Intravenous infusions of CCK-8 (120 pmol/kg/min for 40 min) with vehicle (0 mumol/kg/min proglumide) significantly increased both amylase output in pancreatic juice and plasma insulin concentrations (P < 0.05). Concomitant infusions of proglumide (5-40 mumol/kg/min for 50 min) inhibited both amylase and insulin secretory responses induced by CCK-8 infusion. The antagonistic effects of proglumide occurred in a dose-dependent manner, and proglumide infusion at dose of 20 mumol/kg/min or above simultaneously inhibited CCK-8-induced amylase and insulin responses. In conclusion, although the type of receptor involved is not characterized at present, exogenously infused CCK-8 acts on B cells via a CCK-receptor-mediated mechanism and induces insulin secretion in sheep. PMID- 9406449 TI - Species differences between chickens and rats in chemical properties of adipose tissue lipoprotein lipase. AB - Chemical characterization of chicken and rat lipoprotein lipase (LPL) was carried out following purification of LPL. Molecular weight and isoelectric point of both purified enzymes were determined to be 60 KDa and pH 4, while optimum temperature and pH to yield the maximal activity were about 37 degrees C and pH 8.5. Metallic ions, NaCl and protamine sulfate reduced, and heparin increased, both LPL activities. Michaelis constants for LPLs determined with triolein emulsion as the substrate were 0.98 and 1.57, and those of Vmax were 379.2 and 181.3, in chickens and rats, respectively. Triton WR-1339 caused mixed-type inhibition in rat, but inhibited chicken LPL noncompetitively. In LPLs of chickens and rats, values of Ki were 66.7 and 36.4 with triolein emulsion as the substrate, and 832.4 and 66.0 with respective VLDL as the substrate. These results show species difference between chickens and rats in the affinity to lipoproteins of LPL and inhibition of LPL by Triton WR-1339. PMID- 9406451 TI - Heat increment of feeding in Steller sea lions, Eumetopias jubatus. AB - The heat increment of feeding (HIF) was measured in six captive, juvenile Steller sea lions (Eumetopias jubatus), fed meals of either 2 or 4 kg of herring. HIF was calculated as the post-prandial increase in metabolism above baseline levels, and was measured using open-circuit (gas) respirometry. It averaged 12.4 +/- 0.9% (SE) of ingested energy intake for the 4-kg meal trials, and 9.9 +/- 0.9% for the 2-kg meal size. The effect lasted 8-10 hr for the larger meal size. Metabolism peaked 3.7 hr after feeding, and was 2.13 times higher than baseline levels. For the 2-kg meal size, the effect lasted 6-8 hr, with metabolism peaking 2.8 hr after ingestion at 1.76 times baseline levels. Our estimates of HIF for Steller sea lions are at the lower end of estimates for terrestrial mammals, and are consistent with estimates for other marine mammals. PMID- 9406450 TI - Cloning of chick cellular retinol-binding protein, type II and comparison to that of some mammals: expression of the gene at different developmental stages, and possible involvement of RXRs and PPAR. AB - We cloned chick cellular retinol-binding protein, type two (CRBP II) cDNA and compared it with those of some mammals. The deduced amino acid sequence showed that chick CRBP II was one amino acid greater in size than those of mammals, and the nucleotide sequence of chick CRBP II shared 72%-75% similarity with those of mammals. RNA blot hybridization analysis showed that CRBP II transcript of 0.7 kb was first detected in the duodenum of day-18 embryonic chick, and exhibited a rapid increase during 24 hr around the hatching. Northern blot hybridization also revealed that the transcripts of two types of retinoid X receptors (RXR alpha and RXR gamma) and peroxisome proliferator-activated receptor (PPAR) were expressed in the chick duodenum at hatching. The organ culture of day 16 embryonic chick duodenum showed that the addition of 9-cis retinoic acid in the medium caused a significant increase in CRBP II mRNA levels. In addition, arachidonic acid, from which putative ligands for PPAR were supposed to be generated, was accumulated around hatching in the duodenum. The results may suggest that the abrupt increase of the CRBP II gene expression in the chick duodenum around hatching may be related with RXRs and/or PPAR. PMID- 9406452 TI - An avian sodium-hydrogen exchanger. AB - Intestinal sodium transporters, such as the Na+/H+ exchanger (NHE) are important for Na+ conservation in land birds. In mammals, at least five isoforms of the exchanger, NHEs 1-5, have been cloned, with NHE-1 occurring in epithelial basolateral and nonepithelial cell membranes and NHE-3 being restricted to epithelial apical/brush border membranes. We had demonstrated earlier that chicken intestinal brush border membranes possess NHE activity that functionally resembles mammalian NHE-3. In this study, we used mammalian NHE-1 and NHE-3 probes to examine if chicken enterocytes possess these transporters. Antisera against rat NHE-3 recognized a 97 kDa protein in chicken intestinal brush border membrane, while a NHE-3 cDNA probe failed to recognize any transcript. A NHE-1 antibody failed to recognize any protein in brush border or basolateral membrane, while a NHE-1 cDNA probe recognized a 3.9 kb transcript. Thus, there is more than one NHE isoform in chicken intestine, and our results suggest a novel avian NHE family. PMID- 9406454 TI - Bi-phasic allometric growth of the small intestine, cecum and the proximal, middle, and distal colon of rats (Rattus norvegicus Berkenhout, 1764) before and after weaning. AB - We studied whether the growth of the intestinal tract changes at weaning. We weighted fresh tissue and measured segment length of the small intestine, cecum and proximal, middle, and distal colon of 45 male and 51 female F344/Yit rats (Rattus norvegicus Berkenhout, 1764) of 0 to 428 days old. We performed correlation analysis among tissue weight, segment length and empty body weight after logarithmic transformation of the data. The growth of the small and large intestine was biphasic. Intestinal growth exceeded body growth during the suckling period and slowed down suddenly after weaning. Sexual differences existed in intestinal growth, although much smaller than the difference before and after weaning. These results appear to suggest that the size of intestinal tract of the rat does not adapt to nutritional changes at weaning, but prepare for weaning beforehand. PMID- 9406453 TI - Selective brain cooling reduces respiratory water loss during heat stress. AB - Terrestrial mammals developed several mechanisms to reduce water loss to counteract water shortage. One avenue of water loss is the evaporative heat loss by sweating and panting, which increases with body temperature. Sweating and panting are activated by temperature signals of the body, whereby the brain is the most important site in generating temperature signals. Goats, like other artiodactyls, can cool their brains selectively below the temperature of the trunk core. The aim of the present study is to determine whether an inhibition of the selective brain cooling (SBC) mechanism will increase substantially the respiratory evaporative water loss during heat stress due to the higher brain temperature. The inhibition of SBC was performed by increasing brain temperature experimentally at the same rate as trunk temperature by means of extracorporeal heat exchangers. These experiments without SBC resulted in higher respiratory evaporative water loss compared to experiments with normal SBC. Eighteen experiments in two conscious goats had shown that at a trunk temperature of 40 degrees C the respiratory water loss was reduced on average by 29 g/hr (0.7 l/day) due to the effect of SBC. This amount of water corresponds to about one third of the general water requirements. In conclusion, SBC substantially reduces the water loss in goats during heat stress and consequently improves survival chances during water shortage. PMID- 9406455 TI - [What are the training criteria in gynecologic endoscopy?]. PMID- 9406456 TI - [Is there a place for gene therapy in organ transplantation?]. AB - The major research objectives in organ transplantation are to palliate the lack of organs, to decrease the adverse effects of chronic immunosuppression and to improve medium-term and long-term graft survival. Xenotransplantation and induction of a permanent and specific tolerance to an allograft therefore represent two main lines of research which could partly resolve the problems of organ transplantation. The objective of this article is to evaluate the possible role of gene therapy in the development of xenotransplantation and induction of allograft tolerance. They review the various gene vectors currently available as well as the routes of administration of these vectors specific to transplantation. The place of gene therapy is then evaluated in the context of allo- and xenotransplantation. In allotransplantation, transfection of certain genes of interest into the transplant organ before implantation or into the recipient's immune system is considered. Transfection into the transplant organ of genes coding for immunomodulating cytokines (TGF-beta, IL-4, IL-10, etc.), molecules which block the second signal (CTLA4-Ig) or molecules responsible for apoptosis (Fas/FasL) is discussed. The value of gene therapy in the recipient's immune system consists of transfection onto the recipient's bone marrow cells of genes coding for major histocompatibility system molecules (HLA-DR, DQ, etc.). In xenotransplantation, gene therapy will certainly play a major role in the development of transgenic pigs expressing, on the surface endothelium of their organs, certain human molecules which regulate the activity of complement (CD55, CD59, etc.) or which modify the expression of glycosylated xenoantigens (alpha galactosyl) recognized by performed antibodies. PMID- 9406457 TI - [Should adjuvant or neoadjuvant treatment be administered in esophageal cancer?]. AB - This paper is dedicated to adjuvant treatments in surgery for esophageal squamous cell carcinoma. Randomized studies of neo-adjuvant or adjuvant radiotherapy, chemotherapy or radio-chemotherapy are considered. No trial has yet demonstrated the efficacy of a treatment complementary to surgery. Clinical research must be persued, testing the most effective therapies in non-operable cancers of the esophagus, especially "up-to-date" radio-chemotherapy. PMID- 9406458 TI - [Esophageal perforations and ruptures: a plea for conservative treatment]. AB - OBJECTIVES: To identify the determinants and results on conservative management of oesophageal perforations and ruptures. METHODS: Retrospective clinical review of 34 consecutive patients (mean age: 62 years) treated for cervical (n = 10) or thoracic (n = 24) oesophageal disruption between 1985 and 1996. Causes were: spontaneous rupture (n = 10), instrumental perforation (n = 16), alimentary foreign body (n = 6), and blunt (n = 1) or penetrating trauma (n = 1). The diagnostic delay exceeded 24 hours in 15 cases. RESULTS: A nonoperative management was achieved in 8 patients with no mortality. A conservative surgical treatment was attempted in 23 patients, primary repair in 21 and open drainage in 2, with a 17.4% mortality. Resection (n = 2) or exclusion (n = 1) was performed in 3 patients with no early mortality, but one of them died as result of the subsequent reconstructive operation to restore oesophageal continuity. Overall morbidity was linked to the spontaneous cause of the perforation. Outcome of patients undergoing primary repair was not influenced by the diagnostic delay nor the surrounding sepsis. CONCLUSION: Conservative management should be advocated for the treatment of oesophageal perforations and ruptures, even in case of delayed diagnostiqiagnosis, regardless of the surrounding sepsis and cause of disruption. PMID- 9406459 TI - [Vascular clamping of the liver. Indications and limits]. AB - Control of bleeding during liver surgery is an essential prognostic factor for postoperative morbidity and mortality. Several well defined methods are currently available to ensure vascular occlusion, ranging from selective clamping of a segmental pedicle to total vascular exclusion of the liver. These methods of vascular control each have specific indications. However, they can induce ischaemia of the liver whose functional consequences, such as postoperative liver failure, are particularly severe in the case of prolonged ischaemia, affecting the remaining liver and in the presence of histological or functional alterations of the hepatic parenchyma. Selective methods of vascular control, only affecting the part of the liver to be resected, can be used systematically. In contrast, when the occlusion is not selective, they must be used sparingly, essentially in the case of bleeding from the parenchymal section, adopting the principal objective of the briefest possible total ischaemia. Minimization of bleeding must be weighed up against the consequences of ischaemia on the remaining liver, especially in the case of extensive hepatectomy, prolonged clamping and pathological non-neoplastic liver. PMID- 9406460 TI - [Feasibility and short-term results of the "plug" technique in inguinal hernia. A prospective evaluation]. AB - The aim of this prospective study was to assess the feasibility and postoperative outcome of the "plug" technique in inguinal hernia. One hundred and forty-six consecutive patients were operated for 151 hernias. A plug was applied in 131 cases (86.8%). The Lichtenstein technique was used in 20 cases (13.2%) because of a wide weakness of the posterior wall. Eleven (7.3%) postoperative benign complications occurred. No severe complications were observed and no patient was reoperated. The mean duration of oral analgesia was 2.7 (0-10) days. Mean durations of postoperative hospital stay, time off work and cessation of normal activities were 1.2 (0-4) days, 18.1 (1-37) days and 5.8 (1-18) days, respectively. In conclusion, the "plug" technique is feasible in a wide range of hernias and allows a short hospital stay and an early return to normal activity. PMID- 9406461 TI - [Laparoscopic correction of recurrent gastro-esophageal reflux following laparoscopic fundoplication (4 cases)]. AB - A series of 98 laparoscopic fundoplications, included 7 cases (7.1%) of recurrent gastro-oesophageal reflux. Six of these cases occurred within 12 months of surgery. Four were successfully treated by a second laparoscopic procedure. The mean interval between the initial and corrective operations was 10 months. Factors related to failure were: technical errors, operative inexperience, obesity and the size of the hiatus hernia (when crural closure was not performed). Laparoscopic re-operation to was relatively easy and without mortality but had an increased risk of pleural effusions. The mean length of hospital stay for re-operations was identical to that of initial operations (4 days). No further recurrences were noted after a mean follow-up of one year (280 475 days). We conclude that early failures following laparoscopic fundoplication can be effectively dealt with laparoscopic surgery. PMID- 9406462 TI - [Cost of appendectomy: laparoscopy versus laparotomy. A retrospective study of two series of 114 cases]. AB - This retrospective study was designed to compare the cost of laparoscopic appendicectomy (LA) in patients operated in 1991-92 and open appendicectomy (OA) in patients operated in 1989-90. Patients were matched for sex, ASA score and age into 2 homogeneous series: 114 LA and 114 OA. Costs of accommodation, operation and time off work were calculated by the observed costs method: daily cost of the inpatient unit, hourly cost of the operating room-recovery ward, and the patient's consumption. A mean specific cost was added in the case of LA. A significant difference was observed for operating time, time off work and for the cost of postoperative stay, the operation and time off work and the total cost of the disease. The excess cost of the operation in the case of LA was not compensated by the reduction of the accommodation costs A clinical benefit in terms of reduction of pain and local complications has been reported in the literature. The cost of hospitalisation is higher with LA, but the cost of time off work is decreased. LA provides a clinical comfort in all patients and an economic benefit in patients with a professional activity. PMID- 9406463 TI - [Drainage in thyroid surgery. Is it always a must?]. AB - Drainage in thyroid surgery has been a routine but empirical practice with no scientific evidence to support its benefit. A retrospective review of a personal series of 1789 thyroidectomies over a 3 1/2-year period was conducted. Except for thyroid cancer surgery with lymphadenectomy and large mediastinal goiters requiring sternotomy, no case selection for non-drainage was employed. Patients were stratified only on a chronological basis, according to whether key were drained (n = 575, 1993-1994) or not drained (n = 1214, 1994-1996). Both series included toxic goiters, large plunging compressive goiters, bilateral and redo procedures. Severe life-threatening hematoma requiring reexploration occurred in 5 drained patients (0.9%) and in 5 undrained patients (0.4%). Minor postoperative wound hematoma were conservatively treated in 17 drained patients (2.9%) and 6 undrained patients (1.3%). In our experience, drainage after thyroid surgery may not mandatory provided that the field is completely dry before closure. We therefore, progressively modified our operative strategy in order to improve a meticulous haemostatic technique, considered to be more important than the use of drains. Meticulous surgical technique and obliteration of dead space led us to observe a dramatic decrease of the incidence of hemorrhagic complications, eliminating the need for systematic drainage after thyroid surgery. PMID- 9406464 TI - [Post-bulbar stenosis disclosing duodenal tuberculosis. Apropos of a case]. PMID- 9406465 TI - [Treatment of rectal stenosis by linear stapler following laparoscopic correction of rectal prolapse]. PMID- 9406466 TI - [A case of pseudotumoral form of pulmonary nocardiosis with pleuro-parietal extension]. PMID- 9406467 TI - [Imidazole derivatives, platelet aggregation inhibitors]. AB - Study of the platelet aggregation inhibition has shown the efficiency of compounds with imidazole ring alone or fused. All the compounds resulting of the molecular design starting from these structures has an in vitro activity. We have been able to discuss the correlation existing between activities, toxicities and structures with a particular emphasis to the lipophilicity. PMID- 9406468 TI - [Substituted thio-arylidene thiazolidinones: synthesis and structural study]. AB - The synthesis and physico-chemical properties of fourteen 4-thio-5-arylidene thiazolidine-2-ones and eight 3-(4-bromophenacyl)-4-thio-5-arylidene-thiazolidine 2-ones are described. These products were synthetized by the aldolisation crotonisation reaction between aromatic aldehydes and 4-thio-thiazolidine-2-one followed by N-alkylation of this substituted compounds. PMID- 9406469 TI - [Synthesis and structure of substituted arylazo-imidazolidines and arylidenethiazolidines]. AB - Synthesis and physico-chemical properties of four 3-benzyl or 3-(4-chlorobenzyl) 4-thioxo-5-arylazo-imidazolidin-2-ones, five 3-(4-nitrobenzyl)-5 arylidenethiazolidine-2,4-diones and three 3-(4-phenyl-phenacyl)-4-thioxo-5 arylidenethiazolidin-2-ones have been described. These new products were synthesized by an aldolisation-crotonisation reaction from aromatic aldehydes and 3-substituted thioxothiazolidin-2-ones or thiazolidine-2,4-diones. The arylazo imidazolidine compounds were synthesized by copulation of diazonium ions with imidazolidines. Antimicrobial activity was determined for some compounds. PMID- 9406470 TI - [Synthesis and activity of amide and sulfamide aryl-piperazine derivatives in the central nervous system]. PMID- 9406471 TI - [Study of Lavoisier morphine chlorhydrate stability in different active perfusion systems after reconstitution in different solvents]. AB - The stability of morphine chlorhydrate injectable solutions with no preservative used for drug delivery system (PCA) was investigated. Many concentrations of morphine chlorhydrate were prepared using different solvents and in several containers: PCA cartridges and plastic syringes stored at 37 degrees C. Assays of drug substance and of degradation products were determined at different time within 14 days. In such conditions, morphine chlorhydrate solutions were stable: degradation products were quantitated less than the usual normal i.e. 2% of the theoric concentration of the drug. PMID- 9406472 TI - [Toxocara sp in the playgrounds and sand pits in Besancon. Impact in urban hygiene. Prophylactic measurements for a clean city]. AB - The authors presents the results of parasitology study to determine the prevalence of Toxocara species eggs in 29 playgrounds and sandpits of Besancon. Eggs of Toxocara were found in 25 of sandpits. This results were compared with these found in the world literature. PMID- 9406473 TI - [Ophthalmic preservatives and preparations: reality and perspectives]. AB - Preservatives are used in order to prevent contamination of ophthalmic solutions. Besides their lack of antimicrobial efficacy in certain conditions, they raise number of problems in terms of formulation, stability, interaction with containers. There is also an increasing concern about their deleterious effects in prolonged use on ocular tissues and tear film. Various alternatives have been developed and are analysed in this article. PMID- 9406474 TI - [Importation in France of drugs without authorization for marketing]. AB - With a view to protecting Public Health, French regulations subject the importation into France of not locally registered drugs to specific provisions. In all cases, an administrative authorization is required. The rules are more stringent when the drug is to be administered to patients. In the course of this presentation, the following different situations are considered. the importation for analytical, pharmacological or toxicological studies, the importation for clinical research, the importation by the patient himself, the importation for therapeutical purposes. In the last case, two possibilities exist: the so called "autorisation de cohorte" which permits the importation for a group of patients of a drug for which the studies undertaken for registration are not completed but which allow to assume efficacy and safety of the drug, the authorization given to a named patient which is granted upon request of the doctor when efficacy and safety of the drug are assumed according to current scientific knowledge. The advantages and limitations of these procedures are reviewed. PMID- 9406475 TI - [GC12, marker of cells of mesodermal origin. Value and application to cytodiagnosis of serous effusions]. AB - Monoclonal antibody GC12 was examined for its value in cytopathologic diagnosis. Its sensitivity and specificity in staining reactive mesothelial cells in serous effusions was determined using the indirect immuno alkaline phosphatase anti alkaline phosphatase technique. Smears prepared from 78 serous effusions (15 benign, 54 malignant, 9 atypical) were immunostained with GC12 and five other monoclonal antibodies: KL1 directed against cytokeratins, D33 against desmin, E29 against epithelial membrane antigen, Calam 27 against epithelial surface antigen and NEO 723 reactive with carcinoembryonic antigen. GC12 reacted positively with 80 to 100% mesothelial cells from the 15 benign effusions. Carcinoma cells were negative in 87% of malignant cases. Ovarian carcinomas were specially stained among positive cases. One malignant mesothelioma was positive and one was unreactive with GC12. Moreover, the reactivity of atypical cells in 9 serous effusions with GC12 was helpful to characterize benign and malignant effusions: the immunophenotype GC12-, Desmine-, Calam 27+, EMA+, ACE+ allowed detection of carcinoma cells in 2 cases. It is concluded that GC12 is a good tool for distinguishing carcinoma cells from reactive mesothelial cells in serous effusions. PMID- 9406476 TI - [Mucinous tumors of the ovary. Anatomo-clinical aspects, histological classification, prognostic factors and histogenesis]. AB - In contrast with other malignant tumors, the prognosis of malignant ovarian mucinous tumors has not been improved. In fact, their are diagnosed late and their therapy is often inadequate. The absence of consensus concerning the histological criteria of borderline tumors and mucinous carcinomas has led to diverse classifications which are often not reproducible and are also responsible for heterogeneous results concerning patient survival. Unfortunately, none of the biochemical, molecular biology or cytometric markers which have been studied up until now can be used for differential diagnosis. Another particularity of mucinous tumors consists of their association with other ovarian tumors, mural nodules which are reactive or tumor proliferations and which might influence the prognosis of the mucinous tumor, when their are malignant, and peritoneal pseudomyxoma and appendicular mucocele which give rise to problems related to their relation. PMID- 9406477 TI - [Ultrastructure of the kidney in paraquat-poisoned rats. Comparative study with literature data on man and animal]. AB - This study was designed to assess the effect of paraquat on the rat kidney. The experimental animals used were Wistar rats, a strain selected and maintained at the Institut d'hygiene alimentaire. Their diet was well defined and their state of health monitored. Female animals with mean weight of 200 g were used in this study. Doses of 20, 30 and 50 mg/kg were administered via the gastrointestinal route. In this experimentation, performed on 12 rats plus 3 controls, the dose considered to be sublethal was 30 mg/kg administered by gavage. These animals were sacrificed after 24 h, 48 h, 4, 8, 15, 30 and 60 day, selecting those animals with the most severely altered state, at each time. Tubular lesions started to appear by the 24th hour; the proximal tubule was the most sensitive. Lesions of the distal tubule were observed slightly later, from the 48th hour. Lesions became more intense from the 4th day onwards and reached a maximum on the 8th day. The first signs of repair of the proximal tubule and distal tubule were observed on the 15th day, but were less marked for the proximal tubule. This repair was slow and progressive. Persistent lesions of the proximal or distal tubules were still observed after two months. The glomeruli presented several alterations, which were always only moderate. Overall, paraquat induces serious life-threatening lesions of acute tubular necrosis in the absence of adequate intensive care. PMID- 9406478 TI - [Familial hemophagocytic lymphohistiocytosis. Differential diagnosis with secondary hemophagocytic syndromes]. AB - We report 2 cases of familial hemophagocytic lymphohistiocytosis in two children, heterozygous twins, born from consanguine parents. This disease is characterised by disseminated lymphohistiocytic infiltrates with hemophagocytosis, that most commonly involves bone marrow, spleen, lymph nodes, liver and central nervous system. Differential diagnosis is difficult with infection-induced hemophagocytic syndromes. The only distinguishing feature in pathology is the expression of CD21, CD30 and CD35 antigens by histiocytes. Differenciation is made by an association of clinical and pathologic characteristics: a familial history, lack of infection or neoplasm, and immunohistochemical results. Diagnostic must be rapidly made, because this disease is always fatal without treatment. PMID- 9406479 TI - [Surinfected osteo-chondroplastic bronchopathy. Apropos of a case]. AB - We report a new case of tracheobronchopathia osteoplastica revealed by bronchopulmonary infections. The review of the literature enables us to summarize the clinical, endoscopic, morphological features, as well as pathogenic hypotheses. PMID- 9406480 TI - [Benign anal granular cell tumor. Apropos of a case]. AB - A case of benign granular cell tumor of the perianal region is presented. The patient was treated by local excision of the tumor. This is an unusual site. Superficial biopsy specimens may indicate an incorrect diagnosis of squamous cell carcinoma. PMID- 9406481 TI - Soft tissue angiomyolipoma. A case report. AB - A unique case of angiomyolipoma developed in the deep soft tissues of the lower extremity of a 79 year old woman is presented. The initial tumor was misinterpreted as an ordinary lipoma and the recognition of the entity was accomplished only in the recurrent tumor and after the re-evaluation of the slides from the former tumor. The extensive fibroplasia and the bizarre appearance of many proliferating fibroblasts found in the recurrent tumor have been attributed to local host reaction and degenerative changes. However, these changes may create an alarming histologic pattern to the tumor, which need to be differentiated from a lipo- or leiomyosarcoma. PMID- 9406482 TI - Glandular-type inverted papilloma of the prostatic urethra. AB - A case of inverted papilloma of glandular type in the prostatic urethra of a 65 year-old man is reported. The case was asymptomatic and incidentally discovered on histopathologic study of transurethral resection (TUR) specimens from benign prostatic hyperplasia. The literature concerning this rare entity is reviewed and briefly commented. PMID- 9406483 TI - [Neuroblastoma arising in testicular teratoma]. AB - One case of neuroblastoma arising in an adult immature testicular teratoma is described, with multiple systemic metastases, a partial response to intensive chemotherapy and a swift recurrence leading to death. Such instances of prevailing neuroblastoma with systemic metastases, have only seldomly been reported hitherto. Because of the teratoma and the focal presence of intratubular germ cell neoplasia of unclassified type, we think this tumor must be indeed of germ cell derivation. PMID- 9406484 TI - [Anatomo-clinical criteria of cutaneous melanoma with spontaneous and complete regression. Apropos of a case]. AB - We report a case of a 73 year old man presented a spontaneously and completely regression cutaneous malignant melanoma with cervical lymph node metastasis, confirmed by clinical and histopathologic observations. The authors examine the features of regression of primary malignant melanoma through a review of the literature. PMID- 9406485 TI - Funding research in primary care: is Culyer the remedy? PMID- 9406486 TI - Genetic advances: great promise tempered with concern. PMID- 9406487 TI - Measuring morale--does practice area deprivation affect doctors' well-being? AB - BACKGROUND: Morale is a perennial concern in general practice and, over the years, a variety of tools have been used to examine doctors' mental well-being in a range of psychological and sociological studies. Despite perceived associations between low morale and practice area deprivation, this has not been investigated previously. AIM: To devise and apply a measure of mental well-being in general practitioners, and to use this to investigate the effect of practice area deprivation. METHOD: A questionnaire was devised and piloted, then used in an anonymous postal survey of a random sample of 500 London general practitioners, with questions on demography, workload, practice characteristics, patient centredness, and practice area deprivation. RESULTS: A total of 334 (68%) doctors replied to the questionnaire. Of these, 45% often feel exhausted, 46% are often frustrated by trivial consultations, and a third are seriously disenchanted with work. The resulting well-being score had a normal distribution, was reproducible (test-retest reliability = 0.91), and was internally consistent (Cronbach's alpha = 0.76). Comments from respondents suggested good face validity. Low well-being was not associated with practice area deprivation, but was associated with time stress, small practices and primary care teams, and lack of patient centredness. CONCLUSION: The instrument provided a useful tool for examining doctors' well being and the associations thereof. Well-being was not associated with practice area deprivation. Help for small primary care teams and measures to reduce time stress should help to improve morale. PMID- 9406488 TI - Fundholding in the south Thames Region. AB - BACKGROUND: The general practice fundholding scheme is now at the forefront of the National Health Service (NHS) reforms and should lead to the more efficient use of services by making general practitioners more aware of the financial consequences of their clinical decisions. However, there is a concern that adverse effects may also occur. AIM: To monitor the changes occurring in a sample of fundholding and non-fundholding practices between 1992 and 1995, including providing care nearer to patients, the mixed economy of care, the efficiency and costs of fundholding, and the commitment of fundholders. METHOD: Fifteen first wave practices, four second-wave practices, and four non-fundholding practices in the former South East Thames Region took part in the study. Information was collected using interviews, questionnaires, prescribing data, and annual fundholders' income and expenditure accounts. RESULTS: Consultant clinics were set up in 10 different practices in 15 different specialties, and paramedical clinics in 12 different practices. Physiotherapy and mental health clinics constituted over 90% of the paramedical hours. Fundholders had private arrangements with an individual consultant or practitioner for approximately half of the contracted hours in both types of clinics. Fundholders had lower overall prescribing costs than non-fundholders, but the overall costs for prescribing for all groups had risen by about one third over three years. CONCLUSION: While outreach clinics may help to provide for the needs of patients with common conditions, they may lead to the fragmentation of services. The provision of primary care by those who are not NHS employees needs careful consideration. Recent policies for general practice have emphasized its role in disease prevention and in coordination of care for chronic illness. Fundholding also promotes two additional roles, the purchasing of care and the development of in house facilities. Combining these different functions presents a considerable challenge. PMID- 9406489 TI - Specialist outreach clinics in general practice: what do they offer? AB - BACKGROUND: Specialist outreach clinics in general practice, in which hospital based specialists hold outpatient clinics in general practitioners' (GPs) surgeries, are one example of a shift in services from secondary to primary care. AIM: To describe specialist outreach clinics held in fundholding general practices in two specialties from the perspective of patients, GPs, and consultants, and to estimate the comparative costs of these outreach clinics and equivalent hospital outpatient clinics. METHOD: Data were collected from single outreach sessions in fundholding practices and single outpatient clinics held by three dermatologists and three orthopaedic surgeons. Patients attending the outreach and outpatient clinics, GPs from practices in which the outreach clinics were held, and the consultants all completed questionnaires. Managers in general practice and hospital finance departments supplied data for the estimation of costs. RESULTS: Initial patient questionnaires were completed by 83 (86%) outreach patients and 81 (75%) outpatients. The specialist outreach clinics sampled provided few opportunities for increased interaction between specialists and GPs. Specialists were concerned about the travelling time resulting from their involvement in outreach clinics. Waiting times for first appointments were shorter in some outreach clinics than in outpatient clinics. However, patients were less concerned about the location of their consultation with the specialist than they were about the interpersonal aspects of the consultation. There was some evidence of a difference in casemix between the dermatology patients seen at outreach and those seen at outpatient clinics, which confounded the comparison of total costs associated with the two types of clinic. However, when treatment and overhead costs were excluded, the marginal cost per patient was greater in outreach clinics than in hospital clinics for both specialties studied. CONCLUSION: The study suggests that a cautious approach should be taken to further development of outreach clinics in the two specialties studied because the benefits of outreach clinics to patients, GPs and consultants may be modest, and their higher cost means that they are unlikely to be cost-effective. PMID- 9406490 TI - Do general practitioners and community pharmacists want information on the reasons for drug therapy changes implemented by secondary care? AB - BACKGROUND: The content of discharge prescriptions/summaries to improve communication about medication provided at discharge has been the subject of recent studies. To date, the authors are not aware of any literature that assesses the need for primary care health professionals to receive information on reasons for drug therapy changes incurred during hospital admission. Owing to increased emphasis on seamless care, patient education, and increased accountability for drug costs, general practitioners (GPs) and community pharmacists may consider the receipt of information on the reasons for drug therapy changes incurred during hospital admission to be an essential requirement. AIM: To determine whether GPs and community pharmacists want, and receive, information on the reasons for drug therapy changes implemented by secondary care. The preferred method of acquiring this information is also investigated. METHOD: A questionnaire was posted to all GPs and community pharmacists within the catchment area of Glasgow Royal Infirmary University NHS Trust. Data were collected between June 1995 and July 1995. RESULTS: Replies were received from 71 (64%) GPs and 33 (80%) community pharmacists. Of the respondents, 96% of GPs and 94% of community pharmacists would like information on one or more reason types for drug therapy changes, but the majority do not receive the desired information. Ninety per cent of GPs and 85% of community pharmacists seek this information of facilitate continuity of patient care. The preferred method of receiving the information is by postal delivery via a modified hospital discharge prescription. CONCLUSION: The existing hospital discharge prescription requires modification to facilitate the completion of the reasons for drug therapy changes. The issue of patient-held cards requires consideration. These factors may facilitate continuity of patient care on hospital discharge. PMID- 9406491 TI - Green prescriptions: attitudes and perceptions of general practitioners towards prescribing exercise. AB - BACKGROUND: This qualitative study was part of a broader randomized controlled trial which showed that written exercise advice (green prescription) from a general practitioner (GP) increased physical activity levels among sedentary patients more than verbal advice alone over a 6-week period. AIM: To assess the attitudes and perceptions of GPs towards the practice of writing green prescriptions. METHOD: Participating GPs (n = 25) discussed attitudes and perceptions towards green prescriptions through structured focus groups within 2 weeks of the end of recruitment for the main study. RESULTS: The GPs felt comfortable discussing and prescribing exercise with and to patients. They preferred giving green prescriptions to giving verbal advice alone, and felt they were a valuable tool to formalize and document mutually agreed exercise goals. Time constraints were identified as a major barrier to the widespread implementation of green prescriptions. Appropriate training, resource materials, and patient follow-up mechanisms were identified as important elements for successful implementation of the strategy. CONCLUSION: Overall, the GPs were very positive about the green prescription concept, believing it to be beneficial for patients and achievable within general practice. PMID- 9406492 TI - Obtaining the views of general practitioners on the services to which they refer patients--a locality approach. AB - A survey, with a locality emphasis, of the opinions of Fife general practitioners (GPs) on the quality and availability of a selection of services to which the GPs refer their patients was undertaken. Far more GPs rated services as 'poor' for availability than for quality. GPs acting as locality advisers were actively involved in the planning and execution of the survey as well as the dissemination of the results. The overall response rate was disappointing considering this approach. PMID- 9406493 TI - The Dundee out-of-hours cooperative: preliminary outcomes for the first year of operation. AB - The Dundee out-of-hours cooperative (DDOC) was the first of its kind to be developed in a city in Scotland. In its first year of operation, the key features of the cooperative were that (a) two doctors could deal with the large majority of out-of-hours calls for a population of just under 100,000 patients, (b) only one third of calls required a home visit, (c) most calls were handled within a period of time acceptable to patients, (d) seven out of 10 patients rated the service equal to or better than previous services, and (e) there were extended opportunities for general practitioners (GPs) involved in the scheme to meet with other colleagues during out-of-hours work. PMID- 9406494 TI - Magnetic resonance imaging of the lumbar spine: direct access for general practitioners. AB - At the Cardiff Royal Infirmary we have offered general practitioners (GPs) direct access for magnetic resonance imaging (MRI) of the lumbar spine for sciatica or suspected spinal claudication since January 1993. We compared referrals for MRI from GPs and hospital outpatient doctors, and assessed GP patient management following the scan report. No difference in the diagnostic rates for disc herniation and spinal stenosis were found. GP direct access shortens investigation time, potentially reduces waiting lists, and allows GPs to make more informed management decisions. PMID- 9406495 TI - The evaluation of stress management strategies in general practice: an evidence led approach. AB - Recent surveys have highlighted sources of stress for UK general practitioners (GPs). Interventions to reduce stress in general practice have been introduced at both an individual and an organizational level, but there is little published evidence of their effectiveness. This paper systematically reviews the literature and reports that the research evidence from stress management programmes employed with other workforces is equivocal. Results so far suggest that relaxation and cognitive behavioural skills are helpful and that group methods are both more cost-effective and more beneficial than individual counselling. It is important for scientific, practical, and financial reasons that stress management programmes be properly evaluated. This paper suggests possible avenues for future interventions to alleviate stress. PMID- 9406496 TI - The prevention of psychological morbidity following perinatal death. AB - In recent years, a significant volume of hospital-based literature has been produced about the management of women and their families after a perinatal death. There has also been a considerable amount of work in the voluntary sector which has recognized this as an area of unmet need. The introduction of regional neonatal intensive care units and the shift from secondary to primary care make the development of a structured community-based approach for this group of vulnerable patients increasingly important. This article documents the evidence for high levels of psychological morbidity following perinatal death, reviews a variety of interventions designed to reduce morbidity, and makes some tentative proposals about the key elements of an effective community-based support programme. PMID- 9406498 TI - Evidence based medicine. PMID- 9406497 TI - Improving the treatment of depression in primary care: problems and prospects. AB - Previous work has succeeded in improving the recognition of depression by general practitioners. This is likely to be of most benefit when it results in effective treatment. Factors compromising the effectiveness of pharmacological treatments include non-compliance, non-response, and relapse of depression. Psychological therapies, such as cognitive therapy, are effective and may prevent relapse, but are not available to the majority of depressed patients seen in primary care. Existing evidence demonstrates that primary care staff can be trained in effective psychological interventions for depression, but interventions need to be developed which are sufficiently brief to be incorporated into routine treatment. Consistent provision of information about depression, coping strategies, and sources of support may improve compliance with treatment and subsequent outcome. PMID- 9406499 TI - Evidence based medicine. PMID- 9406500 TI - Antibiotic management of sore throat. PMID- 9406501 TI - Antibiotic management of sore throat. PMID- 9406502 TI - Antibiotic management of sore throat. PMID- 9406503 TI - Sensible drinking: were GPs influenced by the Government report? PMID- 9406504 TI - GP training in dermatology. PMID- 9406505 TI - Minor surgery in general practice. PMID- 9406506 TI - Hormone replacement therapy. PMID- 9406507 TI - Questionnaire response rates. PMID- 9406508 TI - Spread of HIV from non-drug user to non-drug user. PMID- 9406509 TI - [Examination of the vulva and the vagina: from clinics to biopsy]. AB - The purpose of this article is to describe clinical and colposcopic examination of vaginal and vulvar pathologies. Diagnosis tests are described as Hewitt's test, Collins test and epicutaneous test. According to clinical aspect, samples for bacteriological or cytological examinations are performed. Biopsy sites are selected by colposcopy to exclude atypia or malignancy. PMID- 9406510 TI - [Vulvar tumefactions]. AB - Two types of tumefactions can be distinguished: diffuse, without an actually measurable tumour, and circumscribed, most often corresponding to benign tumours. Among diffuse tumefactions, the author diseases prolapse, inguinal hernias, hematomas, oedemas, vulvar varices, abcesses of the vulva and rare cases of elephantiasis. Each case is described and management is discussed. In a second section, where only large tumours can be termed tumescent, the author discusses cystic formations (serous, mucous and epidermal) and solid formations (fibromas, lipomas, acrochordons, neurofibromas, giant condylomas, voluminous endometriomas, and budding neoplastic mass). Excision of these tumours is always recommended as is pathologic surveillance. PMID- 9406511 TI - [Vulvovaginitis]. AB - Candidiasis, infection due to Trichomonas vaginalis and bacterial vaginosis (Gardnerella vaginalis and/or other species) represent the major three causes of vulvo-vaginitis. Other are rare bacterial infections and non infectious vaginitis such as allergic and post-menopausal vaginitis with epithelial atrophy. Clues for the diagnosis include the clinical features of vaginal discharge, cytological examinations, bacterial and fungal cultures. Only T. vaginalis seems to be responsible of sexually transmitted disease. All appropriate antibacterial or anticandidosic treatment are immediately effective, but the mechanisms of recurrent candidiasis and vaginosis are still unclear. PMID- 9406513 TI - [Vulvar dermatoses]. AB - Vulvar dermatoses are the localizations to the vulva of a great variety of dermatoses. In genodermatoses, the expression of diseases on the vulva is a part of a complex syndrome. It is non indispensable for the diagnosis. In the immuno allergic dermatoses or in dermatoses where the pathogenesis is not clearly understood, the vulvar disease is very important because the vulva is often the exclusive localization. The vulvar disease can be the first manifestation before the other expressions of the mucocutaneous disease. Sometimes, there is a risk of scars or transformation in epithelioma. So the diagnosis must be soon established. At least, hidradenitis suppurativa is disease where the choice of the must appropriate treatment is difficult. PMID- 9406512 TI - [Vestibulitis and dyspareunia]. AB - Vulvar vestibulitis syndrome is defined as chronic vulvar erythema involving dyspareunia. Some infectious causes have been suggested, as human papillomavirus. Other known or suspected causes include use of chemicals or other irritants. Sexual active women of the third decade are classically exposed to this syndrome. Dyspareunia is complex with a neurophysiological cause and an emotional outcome. Patients require a medical, surgical or psychological follow-up according to each case. PMID- 9406514 TI - [Vulvar lichen sclerosus]. AB - The most common form of vulvar dermatose is the lichen sclerosus. More frequent after menopause, it can be observed at all ages, even in childhood. Vulvar lichen sclerosus manifests usually by a vulvar pruritus and (or) orificial dyspareunia, however in 15% of cases it remains asymptomatical. The lesions can extend to all the vulva, giving it a very particular pearly white aspect, or remain localized as a "leucoplasia", requiring biopsy to confirm the diagnosis. It is mostly the hyperplastic lichen sclerosus (consisting of a marked epithelial hyperplasia) which risks degeneration. In case of non-response to a strong local steroid therapy, high risk clinical cases must be pointed out, treated and biopsied. A biopsy must be done in thick leucoplastic areas which are coarse, steady and rebellious to treatment, specially on ulcered zones without any tendency to healing or palpable infiltrated lesions. PMID- 9406515 TI - [Vulvar dysplasia]. AB - The histological picture of severe stage dysplasia or undifferentiated vulvar intraepithelial neoplasia III is similar in three clinical entities, which should be distinguished because they differ with regard to clinical characteristics, development, prognosis and treatment. They are: 1. Bowen's disease in the menopausal female, which is a precancerous state that in 10 to 30% of cases develops to invasive epidermal cancer. Surgical excision is required. 2. Bowen's papulosis in the young female, known only since the last 2 decades, is the most frequent. It is clinically highly polymorphic. Since it can regress spontaneously and short- and mid-term malignant transformation is rare, conservative treatment should be given. 3. Extensive, patchy Bowen's papulosis in the young female is associated with intraepithelial cervical neoplasia in 85% of cases and with immune deficiency in 30% of cases. Fortunately, it is rare because its risk of malignant transformation is high. Its treatment has not been fully defined but should be as conservative as possible. This clinical distinction is not accepted in the English literature but to us seems of major importance given the need to apply appropriate treatment in each case. PMID- 9406516 TI - [Cancer of the vulva]. AB - Vulvar cancer develops onto vulvar dystrophies. Its development is linked with HPV infection in half of the cases. It can appear as a carcinoma in situ, a microinvasive carcinoma or a true invasive carcinoma. Prurit is the most common symptom. In situ carcinomas have to be treated by skinning vulvectomies. Radical vulvectomy was considered as mandatory for truly infiltrative cancers. One prefers today use the "wider local excision". However lymphadenectomy is still mandatory (less than 1 mm) in case of very limited dermal infiltration. PMID- 9406517 TI - [Thrombocytopenia. Diagnostic orientation]. PMID- 9406518 TI - [Inhalation corticosteroids. Principles and rules of use]. PMID- 9406519 TI - [Acute respiratory distress syndrome. Etiology, physiopathology, diagnosis]. PMID- 9406520 TI - [Proteinuria (in adults and children). Diagnostic orientation]. PMID- 9406521 TI - [Positive inotropic substances (digitoxin, digoxin). Principles and rules of use]. PMID- 9406522 TI - [Imputability criteria for drug-induced incidents]. PMID- 9406523 TI - [Immunoglobulins E. Role in allergy and atopy]. PMID- 9406524 TI - Identification of the components of simple protein mixtures by high-accuracy peptide mass mapping and database searching. AB - Peptide mass mapping by matrix-assisted laser desorption/ionization (MALDI) followed by database searching with the set of measured peptide masses is now a powerful method for the identification of pure proteins. Protein mixtures--such as frequently occur due to comigration in polyacrylamide gel bands--have hitherto required protein sequencing. Here we demonstrate that such protein bands can also be analyzed by peptide mass mapping alone. Database searching with the complete list of peptide masses determined by delayed-extraction MALDI mass spectrometry with a mass error of less than 30 ppm retrieves the most prominent protein in a mixture. In a second step, the protein identity is further confirmed by matching as many of the measured peptide masses as possible to the retrieved amino acid sequence. Peptide masses remaining after this "second pass search" are searched again to identify the next component in the protein mixture. This iterative process is repeated until all major ion signals are accounted for. Protein mixtures consisting of two or more individual components in a single gel band can be analyzed, further increasing the general applicability of MALDI peptide mapping for protein identification. PMID- 9406525 TI - Molecular imaging of biological samples: localization of peptides and proteins using MALDI-TOF MS. AB - Matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) has been used to generate ion images of samples in one or more mass-to-charge (m/z) values, providing the capability of mapping specific molecules to two-dimensional coordinates of the original sample. The high sensitivity of the technique (low femtomole to attomole levels for proteins and peptides) allows the study of organized biochemical processes occurring in, for example, mammalian tissue sections. The mass spectrometer is used to determine the molecular weights of the molecular in the surface layers of the tissue. Molecules desorbed from the sample typically are singly protonated, giving an ion at (M + H)+, where M is the molecular mass. The procedure involves coating the tissue section, or a blotted imprint of the section, with a thin layer of energy-absorbing matrix and then analyzing the sample to produce an ordered array of mass spectra, each containing nominal m/z values typically covering a range of over 50,000 Da. Images can be displayed in individual m/z values as a selected ion image, which would localize individual compounds in the tissue, or as summed ion images. MALDI ion images of tissue sections can be obtained directly from tissue slices following preparative steps, and this is demonstrated for the mapping of insulin contained in an islet in a section of rat pancreas, hormone peptides in a small area of a section of rat pituitary, and a small protein bound to the membrane of human mucosa cells. Alternatively, imprints of the tissue can be analyzed by blotting the tissue sections on specially prepared targets containing an adsorbent material, e.g., C 18 coated resin beads. Peptides and small proteins bind to the C-18 and create a positive imprint of the tissue which can then be imaged by the mass spectrometer. This is demonstrated for the MALDI ion image analysis of regions of rat splenic pancreas and for an area of rat pituitary traversing the anterior, intermediate, and posterior regions where localized peptides were mapped. In a single spectrum from the anterior/intermediate lobe of a rat pituitary print, over 50 ions corresponding to the peptides present in this tissue were observed as well as precursors, isoforms, and metabolic fragments. PMID- 9406526 TI - Localized sampling of cytoplasm from Xenopus oocytes for capillary electrophoresis. AB - Continued progress in cellular physiology requires new measurement strategies which can be applied to solitary cells. Since many cellular signaling pathways act on time scales of a few seconds, there is a critical need for single-cell techniques with subsecond time resolution. Capillary electrophoresis shows great promise as a tool for the analysis of individual cells. In the present work, we describe a technique to load a capillary with picoliter to nanoliter volumes of cytoplasm and initiate electrophoresis in less than 500 ms. When cytoplasm was sampled from a Xenopus laevis oocyte previously loaded with fluorescein, calcium green, or a mixture of the two fluorophores, their fluorescent peaks were readily identifiable on the electropherogram. Since the volume of cytoplasm (< or = 30 nL) loaded into the capillary was much smaller than the 1 microL oocyte volume, spatially localized biochemical measurements were also possible. To demonstrate the utility of this new technique, the activity of the enzyme beta-galactosidase was measured in small regions of the Xenopus oocyte. Subcellular, subsecond sampling of oocyte cytoplasm will enable biochemical measurements with the resolution required to understand many cellular signal transduction pathways. PMID- 9406527 TI - Detecting biomolecules in picoliter vials using aequorin bioluminescence. AB - The quantitative determination of proteins in picoliter-volume vials is described. The assay is based on the bioluminescence of the photoprotein aequorin along with photon-counting detection. Using this approach, avidin can be detected at femtomole levels by taking advantage of its inhibitory effect on the bioluminescence signal generated by biotinylated recombinant aequorin. The picoliter vials were fabricated on glass substrates using a laser ablation technique. Parameters that affect the reproducibility of the assay such as the fabrication and calibration of the pipets, the fabrication of the vials, and the composition of the assay solutions were studied. PMID- 9406528 TI - A persubstituted cationic beta-cyclodextrin for chiral separations. AB - The applications of a novel polycationic derivative of beta-cyclodextrin (beta CD), heptakis(6-hydroxyethylamino-6-deoxy-beta-cyclodextrin) (beta-CD-EA), as a chiral host--guest additive for the enantioseparation of various classes of chiral anionic analytes are presented. The cationic beta-CD described in this paper is persubstituted with seven ethanolamine side arms at the primary rim of each cyclodextrin (CD) molecule. It is found that the electrophoretic mobility of beta-CD-EA can be adjusted to influence the chiral selectivity by changing the pH of the background electrolyte. Most of the observed CD capillary zone electrophoresis (CZE) separations of anionic drugs and herbicides were accomplished in the pH range of 4.0-7.0 with a reverse polarity configuration. At pH 5.0, enantioseparation of a mixture of three structurally related antiinflammatory agents (fenoprofen, flurbiprofen, and ibuprofen) was possible in about 30 min. However, other chiral acids, such as a series of phenoxypropionic acid herbicides and dansylated amino acids (glutamic acid and aspartic acids), were best separated at pH 6.0 or 7.0. An impressive separation of a mixture of six structurally related anionic herbicides [(+/-)-2-phenoxypropionic acid, (+/-) 2-(2-chlorophenoxy)propionic acid, (+/-)-2-(3-chlorophenoxy)propionic acid, (+/-) 2-(4-chlorophenoxy)propionic acid, (+/-)-2-(2,4-dichlorophenoxy)propionic acid, and (+/-)-2-(2,4,5-trichlorophenoxy)propionic acid] was achieved for the first time in about 15 min during a single run with 20 mM beta-CD-EA. The analytical applicability of this cationic CD molecule for chiral separations is discussed in detail. PMID- 9406529 TI - Fabrication of plastic microfluid channels by imprinting methods. AB - Microfluidic devices have been fabricated on poly(methyl methacrylate) substrates by two independent imprinting techniques. First-generation devices were fabricated using a small-diameter wire to create an impression in plastics softened by low-temperature heating. The resulting devices are limited to only simple linear channel designs but are readily produced at low cost. Second generation devices with more complex microchannel arrangements were fabricated by imprinting the plastic substrates using an inverse three-dimensional image of the device micromachined on a silicon wafer. This micromachined template may be used repeatedly to generate devices reproducibly. Fluorescent analtyes were used to demonstrate reproducible electrophoretic injections. An immunoassay was also performed in an imprinted device as a demonstration of future applications. PMID- 9406530 TI - Effects of ligand heterogeneity in the characterization of affinity columns by frontal analysis. AB - This study examined how the binding capacities and equilibrium constants measured by frontal analysis are affected by ligand heterogeneity in affinity columns. Equations derived for n- and two-site systems gave good agreement with results obtained for the binding of L-thyroxine to a column containing human serum albumin (HSA) and for the binding of (R)-warfarin to coupled columns containing HSA or pigeon serum albumin. The same equations were used to examine how different degrees of ligand heterogeneity affected the apparent binding capacities or equilibrium constants measured using the linear range of double reciprocal frontal analysis plots. A large proportion of two-site systems gave good estimates (i.e., less than 10-20% error) for the true total column capacity and for the association constant of the highest affinity ligand in the column. A smaller, but still appreciable, fraction of all three- and four-site cases also produced good estimates of these values. The results of this work are not limited to protein-based affinity columns but should be applicable to any type of stationary phase that has well-defined binding regions and relatively fast, reversible interactions with solutes. PMID- 9406531 TI - Antibody binding to a functionalized supported lipid layer: a direct acoustic immunosensor. AB - A direct immunosensor has been developed using an acoustic wave device as a transducer. The device is based on an acoustic waveguide geometry that supports a Love wave. The biorecognition surface, formed on a gold layer, consisted of a biotinylated supported lipid layer which specifically bound streptavidin and, subsequently, biotinylated goat IgG. The modified surface was used as a model immunosensor and successfully detected rabbit anti-goat IgG in the concentration range 3 x 10(-8) - 10(-6) M. Using the anti-goat IgG binding isotherm and the time-resolved measurements of antibody binding, both the binding and rate constants of the reaction were determined. The specificity of each binding step was studied with the acoustic wave device, and it was concluded that the phospholipid bilayer showed a good suppression of nonspecific binding. Comparative measurements using surface plasmon resonance allowed the response of the immunosensor to be quantitatively correlated with mass binding to the surface. PMID- 9406532 TI - Ultrasensitive voltammetric detection of underivatized oligonucleotides and DNA. AB - Electrochemical detection of nucleotides, ssDNA, and dsDNA was accomplished by using sinusoidal voltammetric detection at copper microelectrodes. Generally, detection of these molecules utilizes the electroactive nature of adenine and guanine residues at most electrode surfaces. The detection approach used in this study is based on the electrocatalytic oxidation of sugars and amines at copper surfaces. All nucleotides and DNA molecules comprise a ribose sugar backbone and primary amines present on the different nucleobases. Consequently, the detection approach is universal to all types of nucleotides. As the number of sugar moieties increases with the length of an oligonucleotide, the detection sensitivity is enhanced for bigger oligonucleotides. Irreversible adsorption of these oligonucleotides and other biomacromolecules like dsDNA on the electrode surface was avoided with sinusoidal voltammetry since it is a scanning electrochemical technique. The sensitivity of the detection strategy is, however, still preserved due to the effective decoupling of the faradaic signal from the capacitive background currents in the frequency domain. The ssDNA and dsDNA were detected in the picomolar concentration range. The electrochemical signal due to dsDNA is actually higher than that due to ssDNA due to the larger number of easily accessible sugars on the outer perimeter of a dsDNA double helix compared to those on a ssDNA of the same size. This is in contrast to the existing electrochemical detections techniques based on the electroactivity of the nucleobase. PMID- 9406533 TI - A nitrite biosensor based on a maltose binding protein nitrite reductase fusion immobilized on an electropolymerized film of a pyrrole-derived bipyridinium. AB - The preparation and electrochemical characterization of glassy carbon electrodes (GCEs) modified with electropolymerized films of the cation N-(3-pyrrol-1-yl propyl)-4,4'-bipyridine (PPB) are described. The behavior of a new biosensor, which exhibits a high catalytic activity for nitrite reduction and which consists of a maltose binding protein nitrite reductase fusion (MBP-Nir) immobilized on an electropolymerized film of PPB as an electrocatalyst, is also described. The insoluble perchlorate salt of the poly(benzyl viologen) dication was used to immobilize MBP-Nir onto an electrode previously modified with an electropolymerized film of PPB. The electropolymerized film of PPB on the GCE is redox active and exhibits special electron-transfer properties toward the MBP-Nir layer but not toward Nir (Nir without MBP fusion attached), suggesting an intimate interaction between the PPB film and the MBP-Nir layer. The kinetics of the catalytic reaction between the biosensor and nitrite anion were characterized using cyclic voltammetry and rotated disk electrode techniques, and a value of (4.6 +/- 0.5) x 10(3) M-1 S-1 was obtained for the rate constant. PMID- 9406534 TI - Sources of systematic bias in the measurement of water by the coulometric and volumetric Karl Fischer methods. AB - The sources of systematic bias in the measurement of moisture by the volumetric and coulometric Karl Fischer methods were assessed. Using water-saturated octanol as a moisture standard, the measurement accuracy of five coulometric instruments was evaluated. Six possible sources of systematic bias were examined: accuracy of the moisture standard, nonadjustable instrumental bias, operator-adjustable instrumental bias, solvent composition, cell design, and sample composition. The published water content of water-saturated octanol was confirmed by the method of standard additions. The nonadjustable instrumental bias consisted of two types, one that was variable and was observed at water levels below 200 micrograms of water, and one that represented a constant percentage of the total water over the tested range from 40 to 400 micrograms of water. The adjustable instrumental parameters, if set incorrectly, may cause a small but significant negative bias (< 7%). Solvent composition as a function of both solvent type and titration vessel design can introduce a negative bias of up to 10%. Finally, the nature of the sample introduces a negative bias as demonstrated by the results obtained from titrating water in four different oil samples. Under optimum conditions, different amounts of water were measured in each oil using different Karl Fischer titration procedures and five different instruments. PMID- 9406535 TI - A predictive model for matrix and analyte effects in electrospray ionization of singly-charged ionic analytes. AB - In electrospray ionization (ESI), droplets with a surface excess charge are created. The rate of production of surface excess charge is a constant and is equal to the rate of ion production. The ions appearing in the mass spectrum are postulated to be those that formed the surface excess charge at the time of droplet formation (or their collision products). An equilibrium model based on competition among the ions in the solution for the limited number of excess charge sites has been developed. This model accurately predicts the response curves of singly-charged ionic analytes as a function of the concentration of electrolyte and other analytes and provides an explanation for the selective effectiveness of ESI. At low concentrations of total analyte (micromolar and less), the response curves are linear, indifferent to the presence of other low concentration analytes, and suppressed by electrolyte concentrations in excess of the minimum required. At higher analyte concentrations, the response becomes independent of analyte concentration but highly affected by the presence of other analytes. PMID- 9406536 TI - DNA typing of human leukocyte antigen sequence polymorphisms by peptide nucleic acid probes and MALDI-TOF mass spectrometry. AB - A novel analytical method using PNA probes detected by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOFMS) was applied to type sequence polymorphisms within the human leukocyte antigen (HLA), DQA locus. Streptavidin-coated magnetic beads were used to immobilize biotinylated DNA. PNA probes representing possible alleles were then prepared for the immobilized DNA hybridization. The nonspecific PNA probes were removed with stringent washes. The PNA/DNA/beads conjugate was analyzed by MALDI-TOFMS. The genotype of the DNA was determined by the detected molecular masses of the released PNA probes. Reproducible and accurate genotyping was achieved by this analytical method. PMID- 9406537 TI - Release of low molecular weight silicones and platinum from silicone breast implants. AB - We have conducted a series of studies addressing the chemical composition of silicone gels from breast implants as well as the diffusion of low molecular weight silicones (LM-silicones) and heavy metals from intact implants into various surrounding media, namely, lipid-rich medium (soy oil), aqueous tissue culture medium (modified Dulbecco's medium, DMEM), or an emulsion consisting of DMEM plus 10% soy oil. LM-silicones in both implants and surrounding media were detected and quantitated using gas chromatography (GC) coupled with atomic emission (GC-AED) as well as mass spectrometric (GC/MS) detectors, which can detect silicones in the nanogram range. Platinum, a catalyst used in the preparation of silicone gels, was detected and quantitated using inductive argon coupled plasma/mass spectrometry (ICP-MS), which can detect platinum in the parts per trillion range. Our results indicate that GC-detectable low molecular weight silicones contribute approximately 1-2% to the total gel mass and consist predominantly of cyclic and linear poly-(dimethylsiloxanes) ranging from 3 to 20 siloxane [(CH3)2-Si-O] units (molecular weight 200-1500). Platinum can be detected in implant gels at levels of approximately 700 micrograms/kg by ICP-MS. The major component of implant gels appears to be high molecular weight silicone polymers (HM-silicones) too large to be detected by GC. However, these HM silicones can be converted almost quantitatively (80% by mass) to LM-silicones by heating implant gels at 150-180 degrees C for several hours. We also studied the rates at which LM-silicones and platinum leak through the intact implant outer shell into the surrounding media under a variety of conditions. Leakage of silicones was greatest when the surrounding medium was lipid-rich, and up to 10 mg/day LM-silicones was observed to diffuse into a lipid-rich medium per 250 g of implant at 37 degrees C. This rate of leakage was maintained over a 7-day experimental period. Similarly, platinum was also observed to leak through intact implants into lipid-containing media at rates of approximately 20-25 micrograms/day/250 g of implant at 37 degrees C. The rates at which both LM silicones and platinum have been observed to leak from intact implants could lead to significant accumulation within lipid-rich tissues and should be investigated more fully in vivo. PMID- 9406539 TI - Competing unfolding pathways. PMID- 9406538 TI - String and sealing wax. PMID- 9406540 TI - A molecular dipstick? PMID- 9406541 TI - Splitting image. PMID- 9406542 TI - Specific versus non-specific contacts in protein crystals. PMID- 9406543 TI - Pathway of chymotrypsin evolution suggested by the structure of the FMN-binding protein from Desulfovibrio vulgaris (Miyazaki F) PMID- 9406544 TI - Dimerization of the UmuD' protein in solution and its implications for regulation of SOS mutagenesis. AB - NMR spectroscopy has been used to determine that the dimerization interface of UmuD' in solution is not the homodimer interface originally inferred from crystallographic data. Instead, it resembles an interface that had been hypothesized to be involved in filamentation. PMID- 9406545 TI - The lymphoproliferation mutation in Fas locally unfolds the Fas death domain. AB - NMR studies of the lymphoproliferation mutant (V238N) of the Fas death domain indicate that helix 3 is unfolded. This local structural change abolishes binding to FADD--a protein that interacts with Fas and also contains a death domain--and causes the accumulation of autoreactive T cells. PMID- 9406547 TI - Mg(2+)-Mn2+ clusters in enzyme-catalyzed phosphoryl-transfer reactions. PMID- 9406546 TI - A passive transmembrane helix. PMID- 9406548 TI - Crystal structure of the catalytic domain of human phenylalanine hydroxylase reveals the structural basis for phenylketonuria. AB - The 2.0 A crystal structure of the catalytic domain of human phenylalanine hydroxylase reveals a fold similar to that of tyrosine hydroxylase. It provides the first structural view of where mutations occur and a rationale to explain molecular mechanisms of the enzymatic phenotypes in the autosomal recessive disorder phenylketoneuria. PMID- 9406549 TI - Picture story. Carefully coordinated copper chemistry. PMID- 9406550 TI - The structure of mammalian 15-lipoxygenase reveals similarity to the lipases and the determinants of substrate specificity. AB - Here we report the first structure of a mammalian 15-lipoxygenase. The protein is composed of two domains; a catalytic domain and a previously unrecognized beta barrel domain. The N-terminal beta-barrel domain has topological and sequence identify to a domain in the mammalian lipases, suggesting that these domains may have similar functions in vivo. Within the C-terminal domain, the lipoxygenase substrate binding site is a hydrophobic pocket defined by a bound inhibitor. Arachidonic acid can be docked into this deep hydrophobic pocket with the methyl end extending down into the bottom of the pocket and the acid end tethered by a conserved basic residue on the surface of the enzyme. This structure provides a unifying hypothesis for the positional specificity of mammalian lipoxygenases. PMID- 9406551 TI - Structural characterization of activation 'intermediate 2' on the pathway to human gastricsin. AB - The crystal structure of an activation intermediate of human gastricsin has been determined at 2.4 A resolution. The human digestive enzyme gastricsin (pepsin C) is an aspartic proteinase that is synthesized as the inactive precursor (zymogen) progastricsin (pepsinogen C or hPGC). In the zymogen, a positively-charged N terminal prosegment of 43 residues (Ala 1p-Leu 43p; the suffix 'p' refers to the prosegment) sterically prevents the approach of a substrate to the active site. Zymogen conversion occurs in an autocatalytic and stepwise fashion at low pH through the formation of intermediates. The structure of the non-covalent complex of a partially-cleaved peptide of the prosegment (Ala 1p-Phe 26p) with mature gastricsin (Ser 1-Ala 329) suggests an activation pathway that may be common to all gastric aspartic proteinases. PMID- 9406552 TI - Multiple intermediates and transition states during protein unfolding. AB - Rapid kinetic studies of the unfolding of the small protein barstar by urea have been used to demonstrate the presence of at least two unfolding intermediates on two competing unfolding pathways. One intermediate has native-like secondary structure but has a partially solvated hydrophobic core, while the other is devoid of considerable secondary structure but has an intact hydrophobic core. It is shown that the transition states on the two pathways are very dissimilar structurally, but very similar energetically. PMID- 9406554 TI - Crystal structure of horseradish peroxidase C at 2.15 A resolution. AB - The crystal structure of horseradish peroxidase isozyme C (HRPC) has been solved to 2.15 A resolution. An important feature unique to the class III peroxidases is a long insertion, 34 residues in HRPC, between helices F and G. This region, which defines part of the substrate access channel, is not present in the core conserved fold typical of peroxidases from classes I and II. Comparison of HRPC and peanut peroxidase (PNP), the only other class III (higher plant) peroxidase for which an X-ray structure has been completed, reveals that the structure in this region is highly variable even within class III. For peroxidases of the HRPC type, characterized by a larger FG insertion (seven residues relative to PNP) and a shorter F' helix, we have identified the key residue involved in direct interactions with aromatic donor molecules. HRPC is unique in having a ring of three peripheral Phe residues, 142, 68 and 179. These guard the entrance to the exposed haem edge. We predict that this aromatic region is important for the ability of HRPC to bind aromatic substrates. PMID- 9406553 TI - X-ray structure of 5-aminolaevulinate dehydratase, a hybrid aldolase. AB - 5-Aminolaevulinate dehydratase (ALAD) is a homo-octameric metallo-enzyme that catalyses the formation of porphobilinogen from 5-aminolaevulinic acid. The structure of the yeast enzyme has been solved to 2.3 A resolution, revealing that each subunit adopts a TIM barrel fold with a 39 residue N-terminal arm. Pairs of monomers wrap their arms around each other to form compact dimers and these associate to form a 422 symmetric octamer. All eight active sites are on the surface of the octamer and possess two lysine residues (210 and 263), one of which, Lys 263, forms a Schiff base link to the substrate. The two lysine side chains are close to two zinc binding sites one of which is formed by three cysteine residues (133, 135 and 143) while the other involves Cys 234 and His 142. ALAD has features at its active site that are common to both metallo- and Schiff base-aldolases and therefore represents an intriguing combination of both classes of enzyme. Lead ions, which inhibit ALAD potently, replace the zinc bound to the enzyme's unique triple-cysteine site. PMID- 9406556 TI - The future of graduate medical education. PMID- 9406555 TI - A designed four helix bundle protein with native-like structure. AB - A 108 amino acid protein was designed and constructed from a reduced alphabet of seven amino acids. The 2.9 A resolution X-ray crystal structure confirms that the protein is a four helix bundle, as it was designed to be. Hydrogen/deuterium exchange experiments reveal buried amide protons with protection factors in excess of 1 x 10(6) in the range characteristic of well protected protons in functional folded proteins (10(3)-10(8)) rather than protons in rapid exchange (0 10(2)). The protein is monomeric at 1 mM, the concentration at which the exchange experiments were undertaken, indicating that the exchange factors are due to a unique stable tertiary structure fold, and not due to any higher order quaternary structure. Thermodynamic analysis provides an estimate of the free energy of folding of -9.3 kcal mole-1 at 25 degrees C, consistent with the free energy of folding derived from the protection factors of the most protected protons, indicating that global unfolding is required for exchange of the most protected protons. PMID- 9406557 TI - The case of the creeping papules. PMID- 9406558 TI - Fireworks over fibromyalgia, CFS, and IBS. PMID- 9406559 TI - Fireworks over fibromyalgia, CFS, and IBS. PMID- 9406560 TI - Fireworks over fibromyalgia, CFS, and IBS. PMID- 9406561 TI - Fireworks over fibromyalgia, CFS, and IBS. PMID- 9406562 TI - Community-acquired pneumonia. Tailoring management of adult patients according to risk category. AB - A cost-conscious evaluation of CAP in the adult patient requires an initial assessment of the clinical severity and the risks of complicated pneumonia. Initially, most patients should have chest radiography; some authorities also recommend sputum Gram staining and culture, but additional blood testing, culture, and diagnostic procedures are indicated only for patients who have chronically impaired health or clinical evidence of sever infection. Initial empirical antibiotic therapy varies depending on the setting (outpatient, hospitalized patient, critically ill patient). Failure of the patient to respond to empirical antibiotic therapy within 72 hours should direct the clinician to consider unusual or resistant organisms or noninfectious causes of pneumonitis. PMID- 9406563 TI - Rheumatic fever revisited. Keep this diagnosis on your list of suspects. AB - The symptoms of recurrent rheumatic fever in adults can be very different from those in children, and the diagnosis is easy to overlook. Nonetheless, rheumatic fever is still a problem in many parts of the world, as illustrated by this case. Dr Capizzi and associates remind us to keep our options open. PMID- 9406564 TI - Vaccines for hepatitis A and B. The latest recommendations on safe and extended protection. AB - Hepatitis A vaccines (Havrix and Vaqta), administered in two doses, provide long term protection. Target groups include international travelers, children in high risk communities, homosexually active men, injecting drug users, persons who work with nonhuman primates, patients with chronic hepatitis, and recipients of clotting factors. The place of hepatitis A vaccination in the childhood immunization schedule has not been determined. Postexposure prophylaxis for hepatitis A consists of administration of immune globulin within 2 weeks of exposure. Hepatitis B vaccines (Recombivax HB and Engerix-B), administered in three doses, provide protective antibody levels in more than 95% of recipients. Duration of protection appears to approach 10 years. Booster doses are not currently recommended. Hepatitis B vaccination has been incorporated into the routine childhood-immunization schedule. Additional target groups include medical personnel exposed to blood products, household and sexual contacts of infected persons, injecting drug users, and homosexually active men. Postexposure prophylaxis consists of administration of hepatitis B immune globulin as soon after exposure as possible, along with the initial dose of vaccine if desired. PMID- 9406566 TI - Newly diagnosed hepatitis C. Lack of symptoms doesn't mean lack of progression. AB - Hepatitis C is the most common liver disease currently seen in clinical practice. Many patients remain asymptomatic until decompensation occurs. Lack of symptoms and minor liver enzyme elevations are typical of HCV infection and cannot be taken as evidence of lack of progression. HCV RNA testing confirms the diagnosis. Liver biopsy helps assess disease activity and stage the severity of fibrosis and is recommended for most patients with hepatitis C. Once this information is obtained, a rational program for treatment and monitoring can be planned. Patients with newly discovered hepatitis C infection require thorough education about the disease's natural history, transmission, interaction with alcohol, and treatment. In many cases, referral to a gastroenterologist or hepatologist may be the appropriate way to ensure necessary instruction and availability of the latest treatment options. PMID- 9406565 TI - Hereditary hemochromatosis. Preventing chronic effects of this underdiagnosed disorder. AB - Hereditary hemochromatosis, once thought to be rare, is the most common genetic disorder in the United States. Nonetheless, the condition often goes undetected and untreated until its severe effects have become apparent. What clues can lead you to the diagnosis, and how can you spot them in your patients, before significant morbidity has occurred? In this article, Drs McDonnell and Witte discuss the diagnosis and management of this underrecognized problem as well as the various issues involved in screening. An illustrative case of hemochromatosis is also included. PMID- 9406567 TI - Liver transplantation. Who should be referred--and when? AB - Orthotopic liver transplantation is a highly effective treatment for patients with end-stage cirrhosis. At most centers, 5-year survival rates are about 85%. However, donor organs continue to be in short supply, and the issue of patient selection has become increasingly important. The primary care physician plays an important role in referring the patient early--before irreversible complications arise. PMID- 9406568 TI - What the mouth has to say about diabetes. Careful examinations can avert serious complications. AB - Periodontal disease is a major but preventable complication of diabetes mellitus. Patient education, good glycemic control, regular dental care, appropriate diet, and a team approach that involves physicians, dietitians, dentists, and other health professionals offer the best chance for optimum care for these patients. Other oral complications of diabetes include tooth decay, xerostomia, candidiasis, and oral peripheral neuropathy. The mouth may also reflect secondary causes of diabetes, and oral examination may provide clues to diseases that coexist with type 1 diabetes. Truly, the mouth has much to say about diabetes. PMID- 9406569 TI - Calcium channel blockers. Consensus amid controversy. AB - Although the calcium channel blockers have been used to treat hypertension for a number of years, they are now under close scrutiny because of disturbing findings regarding their safety. Drs Kochar and Qurashi sort out the controversies surrounding these widely used drugs and make recommendations based on current consensus. PMID- 9406571 TI - Diagnostic imaging. Finding new ways to see; seeing new ways to cure. PMID- 9406570 TI - Psychosis in late life. Spotting new-onset disorders in your elderly patients. AB - Psychotic disorders usually develop by middle age, but initial onset can be later in life. Delusions, hallucinations, and other psychotic symptoms may accompany medical conditions, mood disorders, and Alzheimer's disease, thereby confusing the diagnosis. In this review, Dr Harris describes some of the more common psychotic disorders in elderly patients and briefly discusses the newest options in drug therapy. PMID- 9406572 TI - Occurrence of BCR-ABL rearrangement in a Philadelphia chromosome-negative patient with 5q and 13q deletions and myeloproliferative syndrome. AB - We report the case of a patient with a myeloproliferative syndrome and traits of myelodysplasia and myelofibrosis whose karyotype showed 5q and 13q deletions, as well as Philadelphia chromosome negativity. A molecular biology study performed by Southern blot, with a probe covering the M-bcr region, led to detection of three bands other than the germinal ones, which hints at the possible existence of two cut points in the M-bcr region of an allele, or participation of both alleles. The patient presented a complex hematological picture, which might be explained on the basis of the cytogenetic and molecular findings. PMID- 9406573 TI - Sublocalization of the breakpoints of a t(5;16) in myelodysplasia. AB - As a first step in characterizing a t(5;16)(q31;p11.2) in a patient with the diagnosis refractory anemia with ring sideroblasts, a cell fusion was carried out between bone marrow cells from the patient and the Chinese hamster cell line A3. Using PCR and FISH analysis on hybrid lines containing the human derivative 16 chromosome, the breakpoints could be mapped between the markers TCF-7 and IL-9 on chromosome 5 and OL-7 and s30A4 on chromosome 16, both regions spanning approximately 1 Mb. Since the breakpoint on 5q has occurred in a region that is frequently deleted in myeloid malignancies, the gene disrupted by this translocation could also be implicated in this aberration. PMID- 9406574 TI - Molecular characterization of the 1p22 breakpoint region spanning the constitutional translocation breakpoint in a neuroblastoma patient with a t(1;10)(p22;q21). AB - To characterize the breakpoint in a neuroblastoma patient with a constitutional rearrangement we have constructed a yeast artificial chromosome (YAC) contig extending approximately 6 Mbp in the chromosome 1p22 region that spans the D1S435 and D1S236 loci. This contig has been confirmed by the coincidence of a number of markers in different overlapping YACs. For several of these YACs the overlap was demonstrated following the isolation and sequencing of end clones from which STS markers were generated. The majority of the YACs have been shown not to be chimeric either through the analysis of somatic cell hybrids or fluorescence in situ hybridization. Following the establishment of the contig we have been able to construct a physical map of the region that incorporates six STS and three newly assigned eSTS markers. The generation of this physical map has allowed the reordering of markers in the genetic linkage map for 1p. The physical order is; tel-D1S435-D1S188-D1S424-D1S236-D1D415- D1S420. With the reordering of D1S435 we have been able to join this contig with another reported previously, thereby generating a well characterized 15 Mbp YAC contig in the 1p22-31 region. The 6 Mbp contig described here spans the chromosome 1 constitutional translocation break-point seen in a patient with a t(1;10)(p22;q21) and who had a stage 4S neuroblastoma. YAC fragmentation has been used to define a 200 Kb region within this contig containing the 1p22 breakpoint. Restriction enzyme analysis demonstrates that there are three NotI sites in this region, one of which lies close to the translocation breakpoint site. PMID- 9406575 TI - A novel myeloid cell line, Marimo, derived from therapy-related acute myeloid leukemia during treatment of essential thrombocythemia: consistent chromosomal abnormalities and temporary C-MYC gene amplification. AB - A novel myeloid leukemia cell line, Marimo, was established from bone marrow cells of a patient with secondary acute myeloid leukemia (AML) that had developed during the treatment of essential thrombocythemia (ET) with busulfan. Karyotype at the ET phase was 46,XX,der(15)t(1;15) (q23;p12-13), but at the blastic phase changed to 44,XX,-5,del(8)(q22), add(17)(p11),+18, psu dic(18;9) (q23;p21) x 2 lacking t(1;15). In Marimo cells, C-MYC gene was temporarily amplified by double minutes (dmin) but disappeared at 33 months, whereas t(10;14;11)(q22;q32;q13) and t(10;14)(q22;q32) were added in vitro psu dic(18;9) x 2 and add(17)(p11) were consistently found throughout the culture. These results suggest that this AML clone is not derived from ET but rather is therapy-related. PMID- 9406577 TI - Development of myeloma and secondary myelodysplastic syndrome from a common clone. AB - We report the case of a 49-year-old woman who presented with a monoclonally IgG kappa expressing myeloma since October 1989. Four years later, after 24 cycles of Melphalan-containing chemotherapy, bone marrow (BM) cells of the patient cytologically revealed myelodysplastic changes for the first time. Cytogenetic examination of the BM obtained in January 1994 showed two clonally aberrant main lines. Each of them represented one of the hematological neoplastic diseases. The quantitatively major clone (MDS-clone) showed a deletion of the long arm of chromosome 7, typical for secondary myeloid disorders. The other clone (myeloma (MM) clone) was characterized by a reciprocal translocation between the short arm of chromosome 8, band q24, a region known to contain the c-myc gene, and the long arm of chromosome 2, band p12, where the Ig kappa gene is located. An unusual finding, however, was that an abnormality of the long arm of chromosome 16 could be detected in both obviously unrelated clones. In the further course of the disease, the MDS and MM clones could be detected, both of them showing cytogenetically a clonal evolution characterized by additional clonal abnormalities. Our data stress the significance of cytogenetics in detecting typical clonal abnormalities in different malignant hematological disorders and in detecting "clonal evolution" as an indicator of the progress of the disease. Moreover, our data suggest that MM and MDS may arise from a common stem cell, which may be characterized by a clonal cytogenetic abnormality. PMID- 9406576 TI - Comparative genomic hybridization study on pooled DNAs from tumors of one clinical-pathological entity. AB - Comparative genomic hybridization (CGH) was performed using DNAs pooled from numerous specimens from tumor categories studied case-by-case. The series of six DNA pools consisted of 28 diffuse centroblastic lymphomas (DCL), 28 gastrointestinal stromal tumors (GIST), 21 primary chondrosarcomas (CS), 17 samples from the Ewing family of tumors (ET), 14 liposarcomas (LS), and 14 mesotheliomas (MS). Losses and gains present in at least 50% of the individual specimens were always detected in the pooled DNAs. The loss of the whole p-arm of chromosome 1 was observed even when the affected proportion of individual specimens was only 25%. Gains were also detected at frequencies lower than 50%, but with a high-level amplification in one or more specimens. In conclusion, the present pooled DNA study revealed the following changes: DCL had a gain at 18q22 qter; GIST had losses at 14 and 22q12, and gains at 5p, 8q22-24, 17q22-qter, and 19q13; ET had gains at 1q and 8q13-qter; LS had gains at 1q21-25 and 12q; and MS had a loss at 9p22-pter. No changes were observed in the CS DNA pool. The results from individual specimens also stressed the importance of these chromosomal regions to the tumorigenesis in the corresponding malignancies. This pooled DNA approach can thus be used for fast screening of recurrent DNA copy number in a specific tumor entity. PMID- 9406578 TI - Enhanced expression in seminoma of human zinc finger genes located on chromosome 19. AB - Six Kruppel-type zinc finger (ZF) genes were cloned from a seminoma cDNA library. One, ZFS-1, showed high sequence homology to the ZNF91 KRAB (Kruppel-associated box) ZF gene family and also the same chromosomal assignment. Interestingly, Northern blot analyses using ZFS-1 and ZNF91 revealed that multiple ZF genes on chromosome 19 were predominantly expressed in seminomas. In addition, the testis and the seminoma showed specific expression of 2.3 kb transcript. Our results suggest that ZF genes on chromosome 19 may be implicated in the development and/or growth of seminomas. PMID- 9406579 TI - A novel BRCA1 mutation in an identical twin pair with similar clinical histories. AB - Factors affecting the penetrance and expression of BRCA1 are not understood. Breast cancer risk and ovarian cancer risk, in general, are known to be associated with non-Mendelian factors. However, whether and how these various factors influence tumor development in BRCA1 mutation carriers is not known. Here we report the breast and ovarian cancer syndrome in an identical twin pair. These female identical twins had remarkably similar clinical histories. Both twins developed histologically similar ovarian cancer in their mid-fifties. One twin was diagnosed with stage III disease and died of refractory metastatic disease. The other twin was diagnosed with stage I disease but ultimately died of recurrent disease. Neither twin developed breast or colon cancer. The twins have both similarities and differences in terms of nongenetic cancer-related risk factors. Results of BRCA1 analysis of DNA from both twins revealed a novel mutation, 2711delA, which resulted in a premature termination at codon 892. This report has intriguing implications concerning the role of genotype in the ultimate penetrance and expression of disease among BRCA1 mutation carriers. PMID- 9406581 TI - Cytogenetic analysis of four angiosarcomas from deep and superficial soft tissue. AB - Angiosarcomas are rare malignant vascular tumors, most commonly found in the skin or superficial soft tissue. We found clonal chromosome aberrations in four short term cultured angiosarcomas. Two cases were diagnosed as epithelioid angiosarcomas, and one as postmastectomy angiosarcoma. Two of the tumors were deep-seated and two were superficial. Angiosarcomas from deep, soft tissue are extremely rare and have never been cytogenetically investigated before. The chromosome number ranged from hypodiploid to hypertriploid. When the results from the present study were combined with data on the four previously reported cytogenetically aberrant angiosarcomas, the most frequently rearranged chromosomes were 5, 7, 8, 13, 15, 20, 22, and the Y chromosome. Recurrent changes, each found in three of these eight angiosarcomas, were gains of 5pter p11, 8p12-qter, and 20pter-q12, losses of 7pter-p15 and 22q13-qter, and -Y in two of three men. PMID- 9406580 TI - De novo AML with trilineage myelodysplasia and a novel t(11;12)(p15;q13). AB - De novo acute myeloid leukemia with trilineage myelodysplasia (AML/TMDS) is an uncommon form of leukemia characterized by a dyshematopoietic picture accompanying the acute leukemia, a poor response to induction chemotherapy, and a tendency to relapse with pure myelodysplastic syndrome. Cytogenetic information on this entity is scarce, although some cases have been reported to be associated with t(7;11)(p15;p15). A 41-year-old woman who had a history of radiotherapy for breast cancer presented with AML/TMDS and was found to have a unique t(11;12)(p15;q13) abnormality. PMID- 9406582 TI - Radiation therapy for breast cancer and clonal chromosome translocations: a fluorescence in situ hybridization study. AB - The frequency of chromosomal translocations was analyzed in skin fibroblast cell cultures derived from irradiated and nonirradiated skin biopsies from five cases of breast adenocarcinoma in women, who had undergone radiotherapy after surgery. The study was performed at the first annual check-up. Chromosomal in situ suppression (CISS) hybridization was performed using metaphase nonisotopic fluorescence in situ hybridization (FISH) with library probes specific for chromosomes 1, 2, 3, 4, 5, 7, 8, 13, 19, and 20. The results showed that the frequency of clonal translocations in metaphases obtained from irradiated areas was significantly higher than in metaphases from the nonirradiated tissue samples. PMID- 9406583 TI - A microsatellite within the MUC1 locus at 1q21 is altered in the neoplastic cells of breast cancer patients. AB - Paired DNA samples from the neoplastic and nonneoplastic cells of 118 patients with the sporadic, nonfamilial form of breast cancer were analyzed for evidence of genetic alteration at a polymorphic microsatellite mapped to intron 6 within the MUC1 gene at 1q21. Two other microsatellite loci, D1S104 and APO-A2, which also map to 1q21, were analyzed as well. The frequency of alteration at the microsatellite within the MUC1 locus was significantly higher than D1S104 or APO A2 (P < 0.001). Analysis by Southern blotting of the VNTR region of the MUC1 gene revealed an amplification of one allele in 34 of 54 informative cases (63%). There was no significant association between these alterations and alterations of the microsatellite within the same locus, suggesting independent mechanisms were responsible for the genetic changes. Microsatellite loci D17S579 at 17q21, the site of the BRCA1 gene, and D18S34 at 18q21-qter, the deleted in colorectal cancer locus, were also analyzed by PCR. Alterations at D17S579 and D18S34 were detected in 18.8% and 6.2% of patients, respectively (P < 0.001, and P < 0.1 relative to the frequency of alteration at D1S104 or APO-A2). A previously described polymorphism of hMSH2 was altered in 16.4% of cases. PMID- 9406584 TI - 10q deletions in metastatic cutaneous melanoma. AB - Cytogenetic and allelic deletion studies have indicated that the loss of distal chromosome 10q may be a frequent and early event in melanoma tumorigenesis. We have studied nine polymorphic markers spanning 56 cM of this region in 27 advanced melanomas and find that half exhibited loss of the entire region, but none had more limited deletions. Because all these tumors had a codeletion of 9p, the 10q deletion event is likely to impair a pathway other than the cyclin dependent kinase-mediated phosphorylation of the retinoblastoma protein. PMID- 9406585 TI - Sister chromatid exchange analysis of human cells carrying retroviral DNA. AB - Chromosomal abnormalities have been seen in human cells following infection with human T lymphotropic virus type I (HTLV-I). Effects on sister chromatid exchange (SCE) in cells carrying HTLV-I related DNA were examined by treatment with N methyl-N'-nitro-N-nitrosoguanidine (MNNG). Peripheral blood specimens were obtained from 16 donors. Cells were processed for SCE analysis and sera were tested for HTLV-I antibodies by Western blot analysis. DNA from fresh lymphoid cells was analyzed by PCR-SSCP for HTLV-I genes. Of 16 donors, eight were HTLV-I seropositive and eight seronegative. Five HTLV-I infected lymphocyte cultures were also analyzed for SCE. Cells were stained immunocytochemically with anti-p19 for the viral expression. By MNNG treatment, SCE was significantly enhanced in cells derived from two seropositive donors with HTLV-I genes. In cells of six seropositive and eight seronegative donors with or without HTLV-I related DNA, SCE frequency was influenced insignificantly. Cells from all persons were negative for the p19 expression. SCE was greatly enhanced by MNNG treatment in cells from three productively infected cultures when compared with SCE in two nonproductively infected cultures. PMID- 9406586 TI - Gliomas in families: chromosomal analysis by comparative genomic hybridization. AB - Gliomas that aggregate in otherwise unremarkable families may have a heritable genetic basis. To determine the spectrum of genetic alterations in glioma susceptible families, we examined tumor DNA from familial cases for regions of chromosomal gain or loss using comparative genomic hybridization (CGH). We compared chromosomal alterations within and among glioma families to those found in sporadic gliomas. A specific chromosomal abnormality common to the tumors of multiple unrelated probands with glioma or a specific chromosomal abnormality common to multiple affected persons in a single glioma-prone family would support the hypothesis of an inherited predisposition to glioma and at the same time identify specific regions of the genome harboring putative glioma susceptibility genes. Tumor DNA from 11 patients from seven families with two or more individuals with glioma was analyzed, including three members of a remarkable family having 10 affected individuals. We found no chromosomal abnormality common to all tumors of all probands nor did we find family-specific abnormalities in two of three glioma-prone kindreds. There were frequent copy number aberrations (CNAs) on chromosomes 7, 10, 19, and the sex chromosomes; other CNAs included +3q(13.3-29), -4q, +5q, -9q34, +12, -13q(21-->33), -15, -16p, +17qter, -18, -21, and -22. Amplifications occurred at +2 7p(11.1-->12), +2 7q(21.2-->33), +2 12q(13.2-->14), and +2 12p(11-->12). Although there were several novel CNAs [ 16p, and +2 12p(11-p12)], none could readily explain the inheritance of these tumors. PMID- 9406588 TI - Oncogenetic map of human genome. PMID- 9406587 TI - Trisomy 10 in acute myeloid leukemia: report of a new case. AB - We report a case of acute myeloid leukemia (AML) with trisomy 10 as the sole abnormality. We have observed this case among 202 de novo and untreated AML examined cytogenetically in our laboratory during the last 10 years. The patient was an adult man diagnosed with AML-M2. An interesting morphologic finding was the presence in light microscopy and ultrastructural studies of prominent giant "Chediak-Higashi-like" granules in some of the leukemic cells. Cell marker studies showed positivity for CD7 and CD33, as seen in the six previously reported cases. The prognosis has been moderate-good, with a survival of 33 months. Trisomy 10 in AML appears to be a rare recurring numeric chromosomal abnormality and probably linked to a myeloblast subset with a CD33+ and CD7+ phenotype. PMID- 9406589 TI - A modular lymphographic magnetic resonance imaging contrast agent: contrast enhancement with DNA transfection potential. AB - A gadolinium-chelated liposomal contrast agent has been prepared, and magnetic resonance imaging (MRI) efficacy has been examined by indirect magnetic resonance lymphography. A lipidic N,N'-dimethylethylenediamine derivative (4) containing a 10,12-diyne-diacyl domain was treated with DTPA anhydride followed by GdCl3 complexation. The complex was confirmed using MALDI spectrometry. An equimolar mixture of the Gd-chelate lipid and a commercially available diyne-PE was formulated as a liposome suspension and irradiated with UV light prior to imaging experiments. Subcutaneous injection of the liposomal gadolinium agent and subsequent MRI of rabbit axillary and popliteal lymph nodes revealed significant contrast enhancement up to 4 h postinjection. To explore the possibility of imaging a DNA transfection event, the gadolinium contrast mixture was formulated with the cationic transfection lipid DOTAP and complexed with the reporter gene encoding luciferase. DNA transfection studies on the NIH3T3 cell line confirmed the transfection activity of the dual-purpose contrast agent and exemplified the potential toward development of an imaging and DNA delivery vehicle. PMID- 9406590 TI - Examination of the role of the acidic hydrogen in imparting selectivity of 7 (aminosulfonyl)-1,2,3,4-tetrahydroisoquinoline (SK&F 29661) toward inhibition of phenylethanolamine N-methyltransferase vs the alpha 2-adrenoceptor. AB - 7-(Aminosulfonyl)-1,2,3,4-tetrahydroisoquinoline (SK&F 29661, 1) is a potent inhibitor of the enzyme phenylethanolamine N-methyltransferase (PNMT, EC 2.1.1.28). In contrast to other inhibitors of PNMT, it is also highly selective toward PNMT in comparison with its affinity toward the alpha 2-adrenoceptor (PNMT Ki = 0.55 microM, alpha 2 Ki = 100 microM, selectivity [alpha 2 Ki/PNMT Ki] = 180). A diverse set of compounds was synthesized and evaluated to probe the role of the acidic hydrogen of the aminosulfonyl group of 1 in imparting this selectivity. Compounds were designed to investigate the effect on selectivity of the acidity of the NH group [the 7-N-methyl (compound 5) and 7-N-(p-chlorophenyl) (compound 4) derivatives of 1], the relative spatial position of the acidic hydrogen [7-(N-(methylsulfonyl)amino)-1,2,3,4-tetrahydroisoquinoline (6) and 7 ((N-(methylsulfonyl)amino)methyl)-1,2,3,4-tetrahydroisoquinoline (8)], or the effect of the substitution of an acidic phenolic group for the aminosulfonyl moiety [1-(aminomethyl)-6-hydroxynaphthalene (23) and 8-hydroxy-1,2,3,4 tetrahydrobenz[h]isoquinoline (9)]. All of the compounds studied displayed lower affinity for PNMT than 1, and nine of the eleven compounds studied showed increased, rather than the desired decreased, affinity for the alpha 2 adrenoceptor. Specifically, compound 4, in which the aminosulfonyl NH group is more acidic than that in 1, showed greatly reduced selectivity on account of increased alpha 2-adrenoceptor affinity as compared to 1 (PNMT Ki = 2.6 microM, alpha 2 Ki = 6.3 microM, selectivity = 2.4). Compound 8, in which the acidic NH group is in the same region of space as that in 1, although the aminosulfonyl group is reversed with respect to the aromatic ring, showed poor PNMT affinity and modest alpha 2-adrenoceptor affinity (PNMT Ki = 330 microM, alpha 2 Ki = 18 microM, selectivity = 0.055). Compound 9, in which a phenolic group is in the same region of space as the acidic NH of 1, exhibited the best alpha 2 adrenoceptor affinity of any of the compounds studied (PNMT Ki = 0.98 microM, alpha 2 Ki = 0.078 microM, selectivity = 0.080). Results from this study suggest that the selectivity of 1 is not solely due to the presence of an acidic hydrogen on the 7-aminosulfonyl group of 1 but is likely also dependent on some other property (e.g. electron-withdrawing character) of the aminosulfonyl group. PMID- 9406591 TI - Screening solution-phase combinatorial libraries using pulsed ultrafiltration/electrospray mass spectrometry. AB - A method is described whereby a family of homologues is synthesized in a one-pot reaction, without isolation or purification, and the reaction mixture is screened using a competitive binding assay based on pulsed ultrafiltration/electrospray mass spectrometry (PUF/ESMS) to tentatively identify those derivatives having the highest affinity for a target receptor. As a model system to test this approach, a synthetic scheme designed to prepare a series of analogues of the adenosine deaminase inhibitor erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA), as diastereomeric mixtures, was carried out. Pulsed ultrafiltration screening of the crude reaction mixture against controls without protein detected protonated molecules corresponding to EHNA-type derivatives and three of its linear, alkyl homologues but did not show protonated molecules for an isobutyl or benzylic EHNA derivative, suggesting the latter was inactive. To verify this conclusion, we prepared E/THNA, the linear homologues, and the benzylic derivative (each as a diastereomeric mixture) and bioassayed them for them adenosine deaminase inhibition index ([I]/[S]0.5). The bioassay results for the individually synthesized analogues were in good agreement with that predicted by the observed relative ion enhancement in the PUF experiments. Thus, the PUF protocol might be used as a general method to quickly provide direction to the chemist in search of drug candidates. PMID- 9406593 TI - 1-[1-2-Benzo[b]thiopheneyl)cyclohexyl]piperidine hydrochloride (BTCP) yields two active primary metabolites in vitro: synthesis, identification from rat liver microsome extracts, and affinity for the neuronal dopamine transporter. AB - 1-[1-(2-Benzo[b]thiopheneyl)cyclohexyl]piperidine hydrochloride (BTCP, 1) and cocaine bind to the neuronal dopamine transporter to inhibit dopamine (DA) reuptake. However, on chronic administration, cocaine produces sensitization, but 1 produces tolerance. Because metabolites of 1 might be responsible for some of its pharmacological properties, we have identified the primary metabolites of 1 produced by rat liver microsomes and determined their affinities for the DA transporter. Five monohydroxylated derivatives (3, 5, 9, 10, 14) and two degradation compounds (15, 16) were identified as metabolites through comparison with synthetic standards in HPLC and GC systems. Standards were obtained utilizing synthetic schemes previously used for the synthesis of phencyclidine metabolites. In vitro, two compounds (3, 5) showed a high affinity for the DA transporter. These active metabolites might be important in the pharmacology of 1. PMID- 9406592 TI - Intensely cytotoxic anthracycline prodrugs: glucuronides. AB - We previously reported the synthesis of a series of doxorubicin analogue prodrugs that give rise to intensely cytotoxic metabolites in the presence of carboxylate esterases. We now report studies on structurally related beta-glucuronide prodrugs that are converted to similar potent metabolites in the presence of beta glucuronidases. These prodrugs were prepared by reductive condensation of daunomycin or doxorubicin with methyl 1-O-[(1'RS)-1'-ethoxy-4'-oxobutyl]-2,3,4 tri-O-acetyl-beta-D- glucopyranosyluronate in the presence of sodium cyanoborohydride followed by base-mediated cleavage of the glucuronate protective groups. The doxorubicin derivatives were isolated in very low yield, most likely because of the inherent base lability of the parent aglycone. By contrast, fairly good yields of the more base-stable daunomycin analogues were obtained. The target daunomycin glucuronide, N-[(4"RS)-4"-ethoxy-4"-(sodium 1"'-O-beta-D glucopyranuronate)butyl]daunorubicin (6a), had a half-life of 30 h when incubated at a concentration of 12 microM in aqueous 0.05 M phosphate buffer, pH 7.4, at 37 degrees C. Under identical conditions in the presence of 197 units/mumol of Escherichia coli beta-glucuronidase, 6a was hydrolyzed with a half-life of 1.7 h. The single metabolite observed was chromatographically identical with that formed from the hydrolysis of N-(4,4-diacetoxybut-1-yl)daunomycin by carboxylate esterases. 6a was approximately 10,000-fold more toxic to human A375 melanoma cells in the presence of E. coli beta-glucuronidase than in the absence of the enzyme. These findings indicate the therapeutic potential of anthracycline glucuronide prodrugs as independent entities or four use in conjunction with enzyme tissue-targeting strategies such as antibody-directed enzyme prodrug therapy (ADEPT) or gene-directed enzyme prodrug therapy (GDEPT). PMID- 9406595 TI - Design and synthesis of benzoic acid derivatives as influenza neuraminidase inhibitors using structure-based drug design. AB - A series of 94 benzoic acid derivatives was synthesized and tested for its ability to inhibit influenza neuraminidase. The enzyme-inhibitor complex structure was determined by X-ray crystallographic analysis for compounds which inhibited the enzyme. The most potent compound tested in vitro, 5 (4-acetylamino) 3-guanidinobenzoic acid), had an IC50 = 2.5 x 10(-6) M against N9 neuraminidase. Compound 5 was oriented in the active site of the neuraminidase in a manner that was not predicted from the reported active site binding of GANA (4) with neuraminidase. In a mouse model of influenza, 5 did not protect the mice from weight loss due to the influenza virus when dosed intranasally. PMID- 9406596 TI - Effect of plasma protein binding on in vivo activity and brain penetration of glycine/NMDA receptor antagonists. AB - A major issue in designing drugs as antagonists at the glycine site of the NMDA receptor has been to achieve good in vivo activity. A series of 4 hydroxyquinolone glycine antagonists was found to be active in the DBA/2 mouse anticonvulsant assay, but improvements in in vitro affinity were not mirrored by corresponding increases in anticonvulsant activity. Here we show that binding of the compounds to plasma protein limits their brain penetration. Relative binding to the major plasma protein, albumin, was measured in two different ways: by a radioligand binding experiment or using an HPLC assay, for a wide structural range of glycine/NMDA site ligands. These measures of plasma protein binding correlate well (r = 0.84), and the HPLC assay has been used extensively to quantify plasma protein binding. For the 4-hydroxyquinolone series, binding to plasma protein correlates (r = 0.92) with log P (octanol/pH 7.4 buffer) over a range of log P values from 0 to 5. The anticonvulsant activity increases with in vitro affinity, but the slope of a plot of pED50 versus pIC50 is low (0.40); taking plasma protein binding into account in this plot increases the slope to 0.60. This shows that binding to albumin in plasma reduces the amount of compound free to diffuse across the blood-brain barrier. Further evidence comes from three other experiments: (a) Direct measurements of brain/blood ratios for three compounds (2, 16, 26) show the ratio decreases with increasing log R. (b) Warfarin, which competes for albumin binding sites dose-dependently, decreased the ED50 of 26 for protection against seizures induced by NMDLA. (c) Direct measurements of brain penetration using an in situ brain perfusion model in rat to measure the amount of drug crossing the blood-brain barrier showed that compounds 2, 26, and 32 penetrate the brain well in the absence of plasma protein, but this is greatly reduced when the drug is delivered in plasma. In the 4-hydroxyquinolones glycine site binding affinity increases with lipophilicity of the 3-substituent up to a maximum at a log P around 3, then does not improve further. When combined with increasing protein binding, this gives a parabolic relationship between predicted in vivo activity and log P, with a maximum log P value of 2.39. Finally, the plasma protein binding studies have been extended to other series of glycine site antagonists, and its is shown that for a given log P these have similar protein binding to the 4-hydroxyquinolones, except for compounds that are not acidic. The results have implications for the design of novel glycine site antagonists, and it is suggested that it is necessary to either keep log P low or pKa high to obtain good central nervous system activity. PMID- 9406597 TI - Cationic lipid-mediated gene delivery to murine lung: correlation of lipid hydration with in vivo transfection activity. AB - A panel of lipidic tetraalkylammonium chlorides has been prepared and screened in studies of both lipid hydration and in vivo mouse transfection. The effect of cationic lipid structure on liposome surface hydration was determined using differential scanning calorimetry. Increases in headgroup steric bulk and the inclusion of cis-unsaturation in the hydrophobic domain led to greater lipid hydration, indicative of a decrease in lipid polar domain associations. Cationic lipids containing hydrogen-bonding functionality in the polar domain exhibited a corresponding decrease in observed lipid hydration, indicative of an increase in lipid polar domain associations. To explore a potential correlation of the hydration data with transfection activity, we examined the in vivo transfection activity of the lipid panel by direct intratracheal instillation of cationic liposome-DNA complexes into BALB/c mice. The more active transfection agents were the lipids that featured headgroup structures promoting close polar domain association in combination with fatty acyl cis-unsaturation. The hydration data suggest that the more effective transfection lipids for mouse lung delivery are those possessing the greatest imbalance between the cross-sectional areas occupied by the polar and hydrophobic domains. PMID- 9406594 TI - (Aryloxy)alkylamines as selective human dopamine D4 receptor antagonists: potential antipsychotic agents. AB - The discovery of a series of novel (aryloxy)alkylamines with selective affinity for the dopamine D4 receptor is described. Target compounds were tested for binding to cloned human dopamine D2, D3, and D4 receptor subtypes expressed in Chinese hamster ovary (CHO) K-1 cells. A number of compounds demonstrated subnanomolar Ki values for binding to the D4 receptor, with several 100-fold selectivities toward the D2 and D3 receptors. Several compounds with combined D3/D4 receptor binding selectivity were also identified. A limited structure activity relationship study of this chemical series is discussed. In a mitogenesis functional assay, the effect of the test compounds on cellular uptake of [3H]thymidine in D4-transfected CHO 10,001 cells was measured and compared to the response of the full dopamine agonist quinpirole. The activity of the compounds varied from full antagonist to weak partial agonist activity (intrinsic activity of 0-19% in comparison to quinpirole). PMID- 9406598 TI - Nonsymmetrically substituted cyclic urea HIV protease inhibitors. AB - A series of nonsymmetrically substituted cyclic ureacarboxamides was synthesized and evaluated for antiviral activity as a function of the inhibition of HIV protease. Selected protease inhibitors were also evaluated for oral bioavailability. The synthesis, pharmacology, quantitative structure-activity relationship (QSAR), and pharmacokinetics for the series will be discussed. PMID- 9406599 TI - A strategy for the incorporation of water molecules present in a ligand binding site into a three-dimensional quantitative structure--activity relationship analysis. AB - Water present in a ligand binding site of a protein has been recognized to play a major role in ligand-protein interactions. To date, rational drug design techniques do not usually incorporate the effect of these water molecules into the design strategy. This work represents a new strategy for including water molecules into a three-dimensional quantitative structure-activity relationship analysis using a set of glucose analogue inhibitors of glycogen phosphorylase (GP). In this series, the structures of the ligand-enzyme complexes have been solved by X-ray crystallography, and the positions of the ligands and the water molecules at the ligand binding site are known. For the structure-activity analysis, some water molecules adjacent to the ligands were included into an assembly which encompasses both the inhibitor and the water involved in the ligand-enzyme interaction. The mobility of some water molecules at the ligand binding site of GP gives rise to differences in the ligand-water assembly which have been accounted for using a simulation study involving force-field energy calculations. The assembly of ligand plus water was used in a GRID/GOLPE analysis, and the models obtained compare favorably with equivalent models when water was excluded. Both models were analyzed in detail and compared with the crystallographic structures of the ligand-enzyme complexes in order to evaluate their ability to reproduce the experimental observations. The results demonstrate that incorporation of water molecules into the analysis improves the predictive ability of the models and makes them easier to interpret. The information obtained from interpretation of the models is in good agreement with the conclusions derived from the structural analysis of the complexes and offers valuable insights into new characteristics of the ligands which may be exploited for the design of more potent inhibitors. PMID- 9406600 TI - Identification of novel farnesyl protein transferase inhibitors using three dimensional database searching methods. AB - Generation of a three-dimensional pharmacophore model (hypothesis) that correlates the biological activity of a series of farnesyl protein transferase (FPT) inhibitors, exemplified by the prototype 1-(4-pyridylacetyl)- 4-(8-chloro 5,6-dihydro-11H-benzo [5,6]cyclohepta[1,2-b]pyridin-11-ylidene)piperidine, Sch 44342, 1, with their chemical structure was accomplished using the three dimensional quantitative structure-activity relationship (3D-QSAR) software program, Catalyst. On the basis of the in vitro FPT inhibitory activity of a training set of compounds, a five-feature hypothesis containing four hydrophobic and one hydrogen bond acceptor region was generated. Using this hypothesis as a three-dimensional query to search our corporate database identified 718 compounds (hits). Determination of the in vitro FPT inhibitory activity using available compounds from this "hitlist" identified five compounds, representing three structurally novel classes, that exhibited in vitro FPT inhibitory activity, IC50 < or = 5 microM. From these three classes, a series of substituted dihydrobenzothiophenes was selected for further structure-FPT inhibitory activity relationship studies. The results from these studies is discussed. PMID- 9406601 TI - Peptidomimetic inhibitors of the human cytomegalovirus protease. AB - The development of peptidomimetic inhibitors of the human cytomegalovirus (HCMV) protease showing sub-micromolar potency in an enzymatic assay is described. Selective substitution of the amino acid residues of these inhibitors led to the identification of tripeptide inhibitors showing improvements in inhibitor potency of 27-fold relative to inhibitor 39 based upon the natural tetrapeptide sequence. Small side chains at P1 were well tolerated by this enzyme, a fact consistent with previous observations. The S2 binding pocket of HCMV protease was very permissive, tolerating lipophilic and basic residues. The substitutions tried at P3 indicated that a small increase in inhibitor potency could be realized by the substitution of a tert-leucine residue for valine. Substitutions of the N terminal capping group did not significantly affect inhibitor potency. Pentafluoroethyl ketones, alpha,alpha-difluoro-beta-keto amides, phosphonates and alpha-keto amides were all effective substitutions for the activated carbonyl component and gave inhibitors which were selective for HCMV protease. A slight increase in potency was observed by lengthening the P1' residue of the alpha-keto amide series of inhibitors. This position also tolerated a variety of groups making this a potential site for future modifications which could modulate the physicochemical properties of these molecules. PMID- 9406602 TI - Assessment of solvation effects on calculated binding affinity differences: trypsin inhibition by flavonoids as a model system for congeneric series. AB - On the basis of molecular models of the interaction between trypsin and a series of seven structurally congeneric bioflavonoid inhibitors, the influence of solvation effects in the calculation of binding free energy differences in congeneric series has been assessed. The models were derived by making use of the X-ray crystal structure of bovine trypsin and the DOCK program, and the complementarity of the interactions between the functional groups of the docked molecules and the binding site region was corroborated independently with the GRID program. Interaction energies calculated for the complexes using molecular mechanics were found to correlate with the experimental inhibitory activities, although the quality of the correlation was dependent on the molecular modeling protocol. To understand why such correlations could be obtained in the absence of an explicit description of solvent effects, the in vitro activities were transformed into binding free energies, and continuum electrostatic theory was used to incorporate solvent effects by approximating them to the electrostatic contribution to the binding free energies. The results of our calculations show that, within this congeneric series, the cost in electrostatic free energy of desolvating both the enzyme binding site and the buried part of the inhibitors (delta Gdesolv) is roughly constant within the series. On the other hand, the electrostatic interaction energy (EeleLR) varies only slightly along the series in comparison with the van der Waals interaction (EVDWLR), and this variation is mostly solvent-independent, i.e., the reaction field energy of the solvent in the bound state (EsrfLR) makes almost a negligible contribution to the binding free energy differences. As a result, differences in binding free energy are dominated by the van der Waals term, while the electrostatic contribution is, to a good approximation, solvent-independent. A similar scenario may account for the good correlations frequently found between ligand activities and ligand-receptor interaction energies derived using plain molecular mechanics, although generality remains to be determined. PMID- 9406603 TI - Synthesis and pharmacological evaluation of triflate-substituted analogues of clozapine: identification of a novel atypical neuroleptic. AB - The trifluoromethanesulfonyloxy (TfO) analogues 3 and 4 of 8-chloro-11-(4-methyl 1-piperazinyl)-5H-dibenzo[b,e][1,4]diazepine (clozapine, 1) and its 2-chloro isomer (iso-clozapine, 2), respectively, were synthesized via their OMe and OH analogues with the conventional synthetic method of the tricyclic dibenzodiazepines and evaluated pharmacologically along with their parent drugs. The binding profile of the 2-OTf analogue (4) is comparable to the binding profile of 1, although the affinity for the dopamine (DA) D2 receptors is higher (IC50 values are 31 nM and 330 nM for compounds 4 and 1, respectively). Interestingly, no notable affinity for muscarinic receptors could be detected in compound 4. On the contrary, the 8-OTf analogue 3 only displayed affinity for muscarinic M1 receptors (IC50 value 35 nM) and no affinity (IC50 value > 500 nM) for the other receptors tested. The 10 mumol/kg sc dose, but not the 10 mumol/kg po dose, of compound 4 stimulated the output of DA. Increases of 80% and 35% in DOPAC output from the dorsal striatum were seen after sc and po administrations of 10 mumol/kg of compound 4, respectively. Doses up to 100 mumol/kg of compound 3 had no effect on either parameter. Doses up to 100 mumol/kg of compound 4 were not cataleptogenic, but significantly decreased apomorphine-induced locomotor activity. In conclusion, compound 4 (GMC1-169) is a new clozapine-like neuroleptic candidate, which is lacking anticholinergic properties and displays a higher potency, as compared to clozapine (1) itself. PMID- 9406604 TI - On the conformation of tiazofurin analogues. AB - Tiazofurin, an important inhibitor of inosine 5'-monophosphate dehydrogenase, has been argued to possess a restricted glycosylic bond due to an energetically favorable intramolecular (1-4) electrostatic interaction between the partial positive sulfur and the negative oxygen of the ribose. This rigidity has been appointed as a plausible cause that leads to activity in the sulfur containing compounds as opposed to the inactive oxazofurin-like analogues (i.e. S is replaced by an oxygen) that lack this favorable interaction. We reinvestigated this notion by using computational methods to report that although the above interaction (or its lack) is likely to contribute to the low-energy conformation of these classes of molecules, the flexibility of the glycosylic bond is ultimately determined by steric interaction of the heteroatoms with the C2'-H and O4' of the ribose. Application of this theory in the design of new analogues is presented as well. PMID- 9406605 TI - Gabor Szabo (1927-1996). PMID- 9406606 TI - Amino acid sequence homology of factor C produced by Streptomyces griseus with regulatory proteins of zinc finger type. AB - Factor C is a regulatory protein produced by Streptomyces griseus 45H. Factor C like antigen can be detected in the most diverse species examined. It is also present in human serum with an average of 219.4 U/ml of narrow dispersion. The level of factor C-like antigen shows an increase in the sera of patients with different hepatic disorders (341 U/ml), some values being 3-7 times higher than normal (600-1500 U/ml). Correlation was found between the elevated antigen levels and the amount of the bilirubin in the sera of patients with liver cirrhosis. Amino acid sequence homology was found between factor C and zinc finger motifs of several known DNA binding proteins. In fact, factor C was successfully bound to and eluted from a Zn2+ affinity column. Our data show that factor C is probably a zinc finger type DNA binding protein. PMID- 9406607 TI - A short GC-rich sequence involved in deletion formation of cloned DNA in E. coli. AB - A spontaneous deletion probably caused by rec A independent recombination between short-direct repeat sequences was observed in pUC19 plasmid carrying a piece of Streptomyces genomic DNA after culturing in liquid medium. The deletion removed an unknown portion of the cloned DNA and the BamHI-EcoRI part of the multiple cloning site with an additional flanking 111 bp from the vector. At the junction a 13 bp GC-rich DNA sequence highly homologous to a known spontaneous deletion hotspot was found. PMID- 9406608 TI - A possible role of the effect of methionine on the activity of aspartokinase in sporulation of a Streptomyces fradiae mutant. AB - In addition to methionine, aspartate may also induce sporulation in the Streptomyces fradiae St3110 mutant that requires methionine for sporulation but not for growth. Partially purified aspartokinase (EC 2.7.2.4.) of the mutant was tested for feed-back control by methionine, threonine, and lysine. Methionine or threonine alone did not have any significant effect but they had a concerted inhibitory effect together that was further increased by lysine. The threonine lysine combination also caused inhibition of the enzyme activity, while lysine alone activated the enzyme. Methionine was also found to partially repress the aspartokinase activity to about one third of the control. The sporulating aerial mycelia induced by aspartate were lacking the characteristic sporulation pigmentation present in the wild type or after induction of spores by methionine. No significant difference in heat resistance between the two types of the spores was detected. The number of spores produced by aspartate was only about one fourth of that after induction by methionine. The data may indicate a role of aspartokinase control by methionine in restoring the normal sporulation, although in addition to this methionine may act by a different mechanism, as well. PMID- 9406609 TI - Purification and characterization of the catalytic subunit of protein phosphatase 1 from Neurospora crassa. AB - Protein phosphorylation is a universal regulatory mechanism in eukaryotic cells. The phosphorylation state of proteins is affected by the antagonistic activities of protein kinases and phosphatases. Protein phosphatases (PPs) can be classified as serine/threonine and tyrosine specific phosphatases. Ser/Thr phosphatases are divided into four subclasses (PP1, PP2A, PP2B, PP2C) on the basis of their substrate specificity, metal ion dependence and inhibitor sensitivity. We were able to detect the activities of all four Ser/Thr protein phosphatases in the mycelial extract of Neurospora crassa. The catalytic subunit of PP1 was purified 1500-fold with a yield of 1.3% using ammonium sulfate-ethanol precipitation, DEAE Sephacel, heparin-Sepharose and MonoQ FPLC chromatography. The protein product was nearly homogenous, as judged by SDS-polyacrylamide gel electrophoresis. The most important properties of the enzyme were the following: /1/ its molecular mass proved to be 35 kD, /2/ it was completely inhibited by inhibitor-2, microcystin and okadaic acid, /3/ it was bound to heparin-Sepharose, and /4/ its specific activity was 2000 mU/mg. These biochemical properties are very similar to those of the homologous enzyme from rabbit muscle and indicate a high level of conservation of PP1 structure during evolution. PMID- 9406610 TI - Lessons to learn from the cell death and heat shock genes of Caenorhabditis elegans. AB - The nematode Caenorrhabditis elegans is an applicable experimental system for simulation of complex biochemical processes of mammalian cells and tissues. The genetic pathway of programmed cell death (PCD) has been partially clarified in the nematode. Analysis of cell death genes of C. elegans led to the conclusion that PCD is conserved in the animal kingdom. Our intention is to study the role of tissue transglutaminase and heat shock proteins in the process of PCD. Tissue transglutaminase is often observed to be induced and activated during the molecular mechanism of PCD. The connection between the heat shock proteins and PCD is not well understood, but it is clear that many apoptosis inducers lead to increased synthesis of heat shock proteins and production of heat shock proteins is coupled with the appearance of apoptosis in numerous experimental systems. Our preliminary observations made by studying cell death mutants of C. elegans we suggest that transglutaminase and heat shock proteins are involved in the death program of the nematode. PMID- 9406611 TI - Rapid detection of Staphylococcus saprophyticus using primer specific PCR. AB - Staphylococcus saprophyticus is one of the most frequently encountered clinically significant members of the coagulase-negative staphylococci. A set of species specific PCR primers was defined for the detection of Staphylococcus saprophyticus. These primers target variable regions (V3 and V6) of the 16S rRNA gene. Primer-specific PCR has potential applications in epidemiological studies and diagnosis of Staphylococcus saprophyticus. PMID- 9406612 TI - Genetic and developmental analysis of mutant Ketel alleles that identify the Drosophila importin-beta homologue. AB - The Ketel gene of Drosophila melanogaster was identified by four KetelD dominant female-sterile mutations and their 27 revertants. The X-ray and the P-induced KetelR alleles delineated the Ketel locus to the 38E1.2-3 cytological position. Although oogenesis proceeds, normally in the KetelD/+ females, embryogenesis comes to a deadlock shortly after fertilization inside the normal-looking eggs of the KetelD/+ females. The KetelD alleles are dominant negative mutations of antimorph type. Cytoplasm of the KetelD/(+)-derived eggs induce lesions when injected into wild-type eggs and the KetelD alleles can be reverted. Zygotes homozygous for loss-of-function (revertant) KetelR alleles die in second larval instar. Analysis of the cold-sensitive Ketel alleles and the genetic interactions between importin-alpha and KetelR mutant alleles indicate an involvement of the Ketel gene product in oo-, embryogenesis and larval life and show interaction of the KETEL protein with different components of nuclear processes. Molecular analysis (to be published elsewhere) confirmed the genetic data and revealed that the Ketel gene encodes the Drosophila homologue of importin-beta, an essential component of nuclear protein import. PMID- 9406613 TI - The physiology and biosynthesis of secondary metabolites. AB - Recently, several excellent reviews appeared /9, 12, 15, 29/ summarizing new findings of the last years and describing the topic in the new interconnections. The signalling systems through which changes in environmental conditions affecting the growth of microorganism are sensed, transmitted and converted into mechanisms controlling the production of antibiotic can now be investigated with the techniques of molecular genetics. Evidence has been accumulated that demonstrated the key roles played by diffusible molecules in regulating cellular differentiation even among prokaryotic microorganisms. This is exemplified by A factor and its analogues, which act as autoregulators for morphological differentiation and secondary metabolism in Streptomyces. In our article we have concentrated on the physiological changes, which can occur during the growth in natural environment or during cultivation under laboratory conditions. Recent evidence for the presence of molecular signalling systems in Streptomyces is reviewed, along with the inherent implications. The constitution of the metabolic type during the first hours of cultivation has been previously reviewed /36/. PMID- 9406614 TI - Treatment possibility of hypercholesterolaemia associated with hypertriglyceridaemia. AB - Thirty patients were investigated with hyperlipoproteinaemia, 15 patients with non-insulin dependent diabetes mellitus (NIDDM) and 15 with primary hyperlipoproteinaemia. The patients took 250 mg acipimox (5-metyl-pyrazine carboxylic acid 4-oxide) 3 times per day for 2 months. Serum examinations were performed before and after 1 and 2 months of acipimox administration. After treatment the cholesterol and triglyceride levels decreased in both groups. Patients with NIDDM had 11% increase of high density lipoprotein-cholesterol (HDL C) level at the end of the first, and 18% increase at the end of the second month, while patients with primary hyperlipoproteinaemia did not change significantly. The low density lipoprotein (LDL) level did not change significantly in either groups of patients. The apolipoprotein A 1 (apo A 1) levels increased significantly during the second month of acipimox administration. Uric acid levels decreased in both groups, but significant change could be detected mainly in the NIDDM group. Serum glucose level did not change in the non-diabetic patients, while it decreased significantly in the NIDDM group. PMID- 9406615 TI - Multimodal electrophysiological examinations in patients suffering from various tumors of the pineal region. AB - Multimodal electrophysiological examinations: blink-, glabella- and masseter reflexes, as well as brain stem acoustic, somatosensory and visual evoked potentials were examined in thirteen patients with clear consciousness suffering from extra-axial, chronic, expanding processes in the tectal region. According to the data, the authors came to the conclusion that several modalities were often required to make a correct diagnosis or to the localization of the space occupying processes. Functional disturbances of the whole of the lower brain stem, but especially of the mesencephalon and of the lower pons were found in cases of expanding processes surrounding the tectum. PMID- 9406616 TI - Presymptomatic diagnosis of familial colon polyposis. AB - The gene responsible for familial adenomatous polyposis (FAP) has recently been mapped, identified and this makes the presymptomatic molecular diagnosis of the disease possible. It can be performed by direct mutation analysis or indirect haplotype analysis. In families where several affected individuals are available the indirect haplotype analysis is the easiest way for performing presymptomatic diagnosis of persons at risk. Among Hungarian families we have performed haplotype analysis using D5S346, a highly polymorphic dinucleotide CA repeat marker located 30-70 kb downstream from APC gene with the combination of restriction endonuclease Rsal site polymorphism. Marker regions were amplified by polymerase chain reaction (PCR) and basen on the above-mentioned polymorphic systems, the haplotype at the APC locus was determined. We believe that haplotype analysis of individuals at risk in large FAP families containing several affected members is a rapid, efficient, and highly valuable method for presymptomatic diagnosis of familial colon polyposis. PMID- 9406617 TI - Muscle, reflex and central components in the control of the ankle joint in healthy and spastic man. AB - In understanding the control of the ankle joint during different motor tasks, we have to investigate at least three components, namely the influence of i) the passive and intrinsic properties of the intact and active muscle system around the joint (termed the non-reflex component), ii) the mechanical importance of the stretch reflex in the stretched and unloaded muscles, and iii) the supraspinal control of the stretch reflex. This thesis is dealing with the importance of the three components in healthy and spastic persons during sitting, standing, and walking. The results are based on stretch reflex and H-reflex measurements from the ankle extensor muscles. During stretch reflex experiments the foot was mounted to a platform (portable during walking) from which the ankle joint torque and the position were measured. To elicit a stretch reflex, the ankle joint was rotated by a strong motor connected to the platform. The mechanical importance of the stretch reflex was investigated by measuring the changes in joint torque. Electrically, the stretch reflex was recorded as the compound muscle action potential through bipolar surface EMG electrodes placed over the soleus muscle. During H-reflex experiments, the tibial nerve was stimulated at the popliteal fossa and the H-reflex recorded over the soleus muscle as during stretch reflex experiments. To investigate how the contractile properties of a muscle in humans depend on the history of activation, we investigated the intrinsic stiffness of the ankle extensors in healthy subjects. At matched background contraction in sitting subjects, a prolonged contraction increased the intrinsic muscle stiffness by 49%. Muscle yielding has been considered especially important for understanding the reflex compensation. We found a general lack of muscle yield and a mechanically important non-reflex stiffness of the ankle extensors showing that non-reflex stiffness is a prominent factor in normal movements of the ankle joint. In both healthy and spastic persons, we found a mechanically strong stretch reflex in the isometric, contracted muscles during sitting. This posed the question; how is the reflex regulated during more functional motor tasks. This was dealt with by studying the H-reflex during isometric ramp contractions and during walking in healthy and spastic persons. In the healthy subjects the H reflex was modulated in consistency with a task dependent control. In the spastic patients the H-reflex lacked a task dependent modulation. In consistency with earlier findings it was suggested that the decreased modulation could have been caused by decreased control of the pre-synaptic inhibition of the Ia terminals or a change in recruitment gain. To test if the stretch reflex behaved as the H reflex, the short latency stretch reflex was investigated during walking. Here we found that the stretch reflex was strongly modulated during a step in healthy subjects as seen for the H-reflex, but when comparing the stretch reflex at matched excitation levels (same background EMGs) during standing and walking, no task-specific reflex modulation was found except the one relating to the excitation level. Therefore, the results emphasise that at least during walking and standing it is not always possible to draw conclusions about the stretch reflex based on observations of the H-reflex. When investigating the modulation of the short latency stretch reflex during walking in spastic patients, we found that the stretch reflex modulation was impaired in spastic patients at least to the extent demonstrated earlier for the H-reflex. The passive stiffness of the ankle joint was at the same time increased in the patients. At matched ankle extensor contraction levels, stretch responses were compared before and after reversible block of the common peroneal nerve and during an attempted, voluntary, fictive dorsiflexion after common peroneal nerve block. (ABSTRACT TRUNCATED) PMID- 9406618 TI - The nigrostriatal system. An experimental slice culture study of the postnatal rat with a description of the pig mesencephalon. PMID- 9406619 TI - Magnitude and pattern of inner and outer hair cell loss in chinchilla as a function of carboplatin dose. AB - We examined the relationship between carboplatin dose and pattern of IHC and OHC loss in five groups of chinchillas treated with carboplatin: (I) single dose, 38 mg/kg, (II) double dose, 38 mg/kg, (III) double dose, 63 mg/kg, (IV) double dose, 75 mg/kg, and (V) double dose, 100 mg/kg. The pattern of IHC loss was relatively uniform along the length of the cochlea with all doses. Average IHC loss increased from approximately 20 per cent in group I to approximately 100 per cent in groups III, IV and V. Average OHC loss was small or negligible in groups I, II, III and IV; only group V consistently showed large (> 40 per cent) OHC losses. OHC loss progressed along a base-to-apex gradient. The dose of carboplatin which reliably destroyed OHCs was four to five times greater than that needed to damage IHCs. PMID- 9406620 TI - The use of internal speech by children with auditory processing problems. AB - In a memory test based on the phonemic similarity effect, and using visually presented homophone and non-homophone word lists, the serial recall of a group of 18 children with central auditory processing disorders (CAPD) was compared with that of a group of 18 normally hearing matched controls. The controls produced more errors on the homophone than the non-homophone list. The CAPD group showed only a slight bias towards more errors on the homophone list. This difference between the groups implied that, as expected, the controls used internal speech and preferred an articulatory- or auditory-rather than a visually-based processing code. The CAPD group, however, showed only a weak articulatory or auditory coding preference. Thus, the use of internal speech seemed poorly developed in the CAPD subjects. PMID- 9406621 TI - A double-blind cross-over study of a non-linear hearing aid. AB - New hearing aids are usually introduced after clinical trials. These are mostly based on subjects reports, in which it is possible that the subject's judgment of the acoustic performance might be influenced by the awareness that it is a new hearing aid which is being investigated. To examine the benefit of a new non linear amplification circuit, a double-blind cross-over study was conducted. Two 'new' hearing aids were developed; they were identical in external appearance and differed only in that one involved ordinary linear amplification while the other employed compressive amplification (the K-amp circuit). Forty-five experienced users with sensorineural hearing loss, aged 60-80 years, used each of the aids for ten weeks, in balanced order. The subjects' need for hearing aid ranged from listening to radio and television to extensive use in all kinds of demanding listening situations. The results, using a structured questionnaire concerning real-life settings, speech reception tests and subject preferences for a particular hearing aid, showed little difference between the two hearing aids. Twenty-three subjects selected the non-linear amplification circuit, 20 subjects preferred the linear hearing aid and two chose to return to their previous aid. No consistent differences between those preferring the linear circuit and those preferring compression were found. It can be concluded that this compression amplification circuit is not significantly preferred to the traditional linear hearing aid. PMID- 9406622 TI - Auditory performances of a 3-4-7 programmable numeric filter hearing aid. AB - We designed a non-portable prototype seven-filter digital auditory hearing aid. For each of the filters, frequency bandwidth, amplification and compression were programmable in order to adapt these parameters to the deaf patient's audiometric characteristics. We compared the hearing improvement it was possible to obtain either with the three-analogue filter auditory prosthesis Triton 3004 hearing aid from Siemens or with our prototype as a function of the number of filters (three, four or seven) and their frequency bandwidth programmability. We tested 21 patients suffering from moderate to severe sensorineural hearing loss. This study allowed us to demonstrate that a seven programmable-width filter strategy seems to be more effective than the present analogue T004 device. Further studies with improvement of our prototype and finer audiometric adjustment of filter strategies, together with long-term clinical studies, need to be carried out. PMID- 9406623 TI - Concerns over landmark Vet Bill. PMID- 9406624 TI - What's in a name? Veterinary acronyms. PMID- 9406625 TI - A 6-year-old rottweiler with weight loss. PMID- 9406626 TI - Canine histiocytic ulcerative colitis. AB - OBJECTIVE: To assess prevalence, breed predilection, response to therapy and prognosis of canine histiocytic ulcerative colitis (CHUC). DESIGN: A retrospective study of cases of CHUC seen at Sydney University Veterinary Teaching Hospital (SUVTH) over a 20-year period (1975-1994). PROCEDURE: Case records of all dogs in which colitis was suspected were reviewed. Dogs were diagnosed with CHUC based on colonorectal biopsy and histopathological examination. RESULTS: CHUC was diagnosed in 8 of 57 dogs presented for colitis. All affected dogs were Boxers and six were female. Dogs with milder clinical signs showed moderately good response to therapy. Four dogs were still alive 1 to 7 years after diagnosis. Three dogs were euthanased, one for reasons other than CHUC, and one was lost to follow-up. CONCLUSIONS: Young boxer dogs with relatively mild signs of CHUC may respond moderately well to medical and dietary therapy with fair prognosis. Euthanasia early in the course of the disease may be unwarranted. Prevalence may be increasing within the SUVTH referral population. PMID- 9406627 TI - Transient glucose malabsorption in two horses--fact or artefact? AB - Two horses, presented for investigation of chronic weight loss despite normal to increased feed intake, had flat oral glucose absorption curves, suggesting malabsorption. The cause of the apparent malabsorption was not evident grossly or on light microscopic examination of the intestinal tract. Both horses survived long term and at follow-up examination had regained weight and their capacity to absorb glucose. These cases illustrate that flat glucose absorption curves may occur in horses with no obvious intestinal lesions, that they may revert to normal and that the results of these tests should be interpreted with caution. PMID- 9406629 TI - An outbreak of malignant catarrhal fever in young rusa deer (Cervus timorensis). AB - On the basis of clinical signs and histological findings eight 9-month-old male rusa deer (Cervus timorensis) were diagnosed with sheep associated-malignant catarrhal fever. Following a variable course involving rectal temperatures around 40.5 degrees C, depression, inappetence, diarrhoea, corneal opacity and hypopyon all animals died or were euthanased over a 5-week period. Severe multifocal vasculitis, mainly periglomerular and in the arcuate vessels were consistent histological findings which in the past have been adequate to confirm clinical diagnosis of sheep associated-malignant catarrhal fever. A nested polymerase chain reaction test has been used to detect a sheep associated-malignant catarrhal fever PRC product, 238 base-pairs in size, in DNA extracted from lymphocyte preparations. The result supported the diagnosis of sheep associated malignant catarrhal fever in these deer. PMID- 9406628 TI - Thiamine deficiency in a cat associated with the preservation of 'pet meat' with sulphur dioxide. AB - A cat with allergic dermatitis was fed a diet of fresh meat and a multi-vitamin supplement for 38 days to exclude food allergy as a cause of its dermatopathy. The cat died as a result of acute thiamine deficiency, which was caused by inactivation of thiamine by the preservative, sulphur dioxide. The continuing undeclared usage of sulphites in the Australian pet food industry is discussed. PMID- 9406630 TI - Improving the profits of wool-growing farms. PMID- 9406631 TI - Changes in productivity and profitability of wool-growing farms that follow recommendations from agricultural and veterinary studies. AB - OBJECTIVE: To estimate the changes in productivity and profitability in a group of wool-growing farms as they adopted major recommendations from agricultural and veterinary studies. FARMS: Four wool-growing farms in south western Victoria were selected from the clients of the Mackinnon Project, a farm consultancy service run from the University of Melbourne. Each farm had closely followed recommended procedures, kept comprehensive financial and physical records and had been clients for at least 5 years. The comparison group was the South Western Victoria Monitor Farm Project (SWVMFP), about 45 farms in the same region as the study farms that were monitored annually by Agriculture Victoria. PROCEDURE: For a 7 year period, the financial and physical performance of both groups of farms was estimated. Stocking rate, wool production, gross farm income, farm operating costs, net farm income and return on assets were compared. RESULTS: Mean gross farm income of the four study farms steadily rose from 86% of the average SWVMFP farm before the adoption of recommendations to an average of 155%. During the same period, net farm income rose from 70% to 207% of the average of the SWVMFP. Return on asset of the four farms rose irregularly from 26% to 145% of the average of the SWVMFP. Farm operating costs on the four farms were higher than for the SWVMFP group, but the ratio of costs remained relatively constant. CONCLUSION: The adoption of proven research results was associated with large increases in net farm income. An increase in gross income, rather than a reduction in costs was the main reason for this. Research results offer a way to increase the financial viability of wool-growing farmers, many of whom are currently unable to maintain their lifestyle, resources and infrastructure. PMID- 9406632 TI - Sickness, mortality and the buller steer syndrome in a western Canadian feedlot. AB - OBJECTIVE: To determine if an association existed between sickness, mortality and bullers in a western Canadian feedlot. DESIGN: A retrospective epidemiological study. ANIMALS: 78,445 male cattle that entered a 24,000-head feedlot in western Canada from 1991 to 1993. PROCEDURE: Animal health records for bullers were collected and analysed to see if they were at greater risk of sickness and mortality than other steers, and to see if pens with a high prevalence of bullers also had a high prevalence of sickness and mortality. RESULTS: The prevalence of bullers increased with increasing age of cattle on arrival at the feedlot (R = 0.36; P < 0.001). Sickness and mortality decreased with increasing age of cattle on arrival. However, sickness and mortality in bullers relative to other steers actually increased with increasing age on arrival suggesting an interaction existed between sickness and bullers. Bullers were significantly (P < 0.05) more likely to get sick and to die than other steers. In all cases, there was a strong temporal association between sickness and bullers, with sickness and bullers mostly occurring within the first 30 days of the feeding period. On average, pens of cattle with a high prevalence of bullers did not have a correspondingly high prevalence of sickness or mortality. CONCLUSION: This study suggests that sickness is an effect modifier of dominance behaviour and therefore bullers in feedlot steers. Bullers should always be checked for signs of sickness and treated accordingly. Further research is needed to investigate the effects of sickness on dominance behaviour in pens of feedlot cattle. PMID- 9406633 TI - Observations of fibropapillomatosis in green turtles (Chelonia mydas) in Indonesia. AB - OBJECTIVE: To determine the prevalence and manifestations of fibropapillomatosis in green turtles in Indonesia, to identify any relationship between fibropapillomatosis and concurrent parasitic infection, to ascertain the effect of fibropapillomatosis on health, and to examine whether environment might have an effect on the prevalence of fibropapillomatosis. PROCEDURE: 4407 green turtles (Chelonia mydas) and 401 hawksbill turtles (Eretmochelys imbricata) were examined. The occurrence of fibropapillomatosis was correlated with sex, maturity, curved carapace length, body weight/curved carapace length ratio, the number and distribution of tumours on the skin, parasite burdens, some haematological variables and the region of capture. RESULTS: Fibropapillomatosis was seen only in green turtles, and the overall prevalence in these was 21.5%. This prevalence increased with the curved carapace length up to 85 cm. The average number of tumours per affected turtle was 5 +/- SD 4.1 (range, 1 to 29), and was negatively correlated with the body weight/curved carapace length ratio (rs = -0.8; P = 0.001). The red blood cell count in turtles with fibropapilloma was lower than in non-fibropapilloma turtles captured and examined at the same time (P = 0.001). The prevalence of fibropapilloma in turtles captured near densely populated, industrial regions (26.3%) was greater than in turtles from sparsely populated areas (17.7%). CONCLUSION: Fibropapillomatosis in green sea turtles in Indonesia is of moderate occurrence: young mature turtles (curved carapace length = 85 cm) are most frequently affected. Fibropapilloma adversely affects health of turtles. Fluke infestation seems not to be a causal factor, but viral infection, perhaps with concurrent stress of environmental origin, seems likely. PMID- 9406634 TI - Evaluating the risk of the establishment of screwworm fly in Australia. AB - OBJECTIVE: To investigate probabilities of establishment of screwworm fly throughout the year, for several locations around Australia's coastline. METHODS: A simulation model that predicts the spread and economic impact of an established screwworm fly population was modified to include stochastic survival functions, to investigate the risks of the pest actually establishing in this country. The effects of time of year, climate, vegetation and the number of incoming flies or larvae were investigated for four locations around Australia. RESULTS: Analysis of variance identified a dominant three-way interaction between site, time, and the number of introduced flies. These probabilities are graphed. DISCUSSION: In southern areas, as exemplified by Fremantle, the cold winters limit survival. A high probability of establishment exists year round in tropical regions, except in areas around the Gulf of Carpentaria and in the Northern Territory where dry weather mid-year would limit survival. Despite these comparatively lower risks, there were no areas or times where reductions in quarantine efforts could be justified. PMID- 9406635 TI - Postmortem lesions in the intercarpal ligaments of the equine midcarpal joint. AB - OBJECTIVE: To determine the frequency of damage to the medial palmar intercarpal ligament (MPICL), and the range of sizes of the dorsomedial intercarpal ligament (DMICL) of the midcarpal joint in horses with no history of carpal joint disease. MATERIALS AND METHODS: Cadaver limbs were collected from 72 horses with no history of carpal joint disease. One hundred and forty-two midcarpal joints were dissected and the MPICL and DMICL were examined. Measurements were made with a digital micrometer. RESULTS: MPICL tearing was present in 88 of 96 joints from horses 2 years and older. Tears were predominantly of the dorsolateral bundle and complete rupture of the dorsolateral and dorsomedial bundles was not observed. Tearing was not present in foals less than 4 months of age and the severity of tearing increased significantly with age (P < 0.0001). Severity of tearing was significantly greater in racing Standardbreds than racing Thoroughbreds (P < 0.01), but there was no significant difference between racing and non-racing horses. The lateromedial thickness of the DMICL ranged from 0.4 mm to 2.6 mm in horses 2 years and older. Lateromedial thickness increased significantly with age, and was significantly greater in racing Standardbreds than racing Thoroughbreds (P < 0.01). There was no significant difference between racing and nonracing horses. CONCLUSIONS: Damage confined to the dorsolateral bundle of the MPICL is a common finding in horses over 1 year of age and is probably of little clinical significance. Complete rupture of both dorsolateral and dorsomedial bundles is uncommon in horses with no history of midcarpal joint disease. Variation in size of the DMICL is observed in horses of all ages, but is most marked in 2-year-old horses. PMID- 9406636 TI - Effect of increasing the banking of a racetrack on the occurrence of injury and lameness in standardbred horses. PMID- 9406637 TI - Immunohistochemical and microbiological study of the oropharyngeal region of goats experimentally infected with Mycoplasma mycoides cluster strains. PMID- 9406638 TI - Collection of endometrial cells in the mare. PMID- 9406639 TI - Modes of treatment. PMID- 9406640 TI - Modes of treatment. PMID- 9406641 TI - Modes of treatment. PMID- 9406642 TI - Stop the 'uni' bashing. PMID- 9406643 TI - A lacertilian dorsal retinorecipient thalamus: a re-investigation in the old world lizard Podarcis hispanica. AB - The aim of this work is to delineate the retinorecipient cell groups of the dorsal thalamus of lizards and to study some of the differential connections in order to help to understand the evolution of the visual system in tetrapods. Tract-tracing and immunohistochemical techniques were applied to the retinorecipient dorsal thalamus of the lizard Podarcis hispanica. The retina of Podarcis projects to four areas of the dorsal thalamus: nucleus ovalis (Ov), intergeniculate leaflet (IGL), dorsal lateral geniculate nucleus (GLD) and dorsolateral anterior nucleus (DLA). Nucleus ovalis shows a clear cell plate/neuropile organization and projects to the ventral thalamus. Thus, it seems to belong to the ventral rather than to the dorsal thalamus. The IGL contains large cells reactive for GABA and/or NPY immunohistochemistry. It is interconnected with the supra/retrochiasmatic hypothalamus and projects to the opposite thalamus and to the ipsilateral tectum. The caudal DLA, which lacks both GABA- and NPY-like immunoreactive cells, is reached by a few thin retinal fibers, although distal dendrites of DLA cells enter the GLD, suggesting an important retinal input. The DLA projects to the medial and dorsal telencephalic cortices. The GLD is the main retinorecipient thalamic structure that projects to the telencephalon. It shows a crude laminar organization in which cell plate neurons project to the ipsilateral pallial thickening, but it does not receive a descending projection from the visual telencephalon and thus differs from the GLD of other amniotic vertebrates. In the context of present knowledge, these results suggest that an IGL homologue is present in all tetrapods studied, whereas Ov seems to be restricted to diapsid vertebrates. Moreover, our data suggest that a unimodal visual projection to the telencephalon (arising from the GLD) first appeared in reptiles by segregation from a limbic (multimodal) thalamo telencephalic pathway. PMID- 9406644 TI - Tubular eyes of deep-sea fishes: a comparative study of retinal topography. AB - The world's deep oceans are home to a number of teleosts with asymmetrical or tubular eyes. These immobile eyes possess large spherical lenses and subtend a large binocular visual field directed either dorsally or rostrally. Derived from a lateral non-tubular eye, the tubular eye is comprised of a thick main retina, subserving the rostrally or dorsally directed binocular visual field, and a thin accessory retina subserving, the lateral, monocular visual field. The main retina is thought to receive a focussed image, while the accessory retina is too close to the lens for a focussed image to be received. Several species also possess retinal diverticula, which are small evaginations of differentiated retina located in the rostrolateral wall of the eye and thought to increase the visual field. In order to investigate the spatial resolving power of these retinae (main, accessory and diverticulum), the distribution of cells within the ganglion cell layer was analysed from retinal wholemounts and sectioned material in ten species representing four genera. In all species, the main retina possesses a marked increase in cell density towards a specialised retinal region (area centralis), with a centro-peripheral gradient range between 7:1 and 60:1 and a peak density range of between 30 and 55 x 10(3) cells per mm2. The accessory retinae and the transitional zone between the main and accessory retinae possess relatively low cell densities (between 1 and 10 x 10(3) cells per mm2) and lack an area centralis. Retinal diverticula examined in four species possess mean ganglion cell densities of between 7.2 and 109.4 x 10(3) cells per mm2. Analyses of soma areas show that the ganglion cell layer of most species possesses cells with areas in a range of 8.0 to 15.4 microns2 in the main retina and between 15.1 and 17.4 microns2 in the accessory retina. The peak spatial resolving power of the main retina of the ten species varies from 4.1 to 9.1 cycles per degree. The positions of the retinal areae centrales relative to each species' binocular visual field are discussed in relation to what is known of feeding behaviour of these fishes in the deep-sea. PMID- 9406645 TI - Anti-Zebrin II immunopositivity in the cerebellum and octavolateral nuclei in two species of stingrays. AB - Anti-Zebrin II is an antibody directed against a 36kDa aldolase epitope expressed by Purkinje cells. Two patterns of Zebrin II immunolabeling have been described in the cerebellar corpus. In mammalian cerebella, the anti-Zebrin II labels longitudinal zones of Purkinje cells, whereas in teleosts, all Purkinje cells of the cerebellar corpus are Zebrin II immunopositive. An outgroup analysis is required to determine which of these distribution patterns represents the primitive condition for jawed vertebrates. The sister group of the Osteichthyans (rayfinned fishes, amphibians, and amniotes) is the Chondrichthyans (sharks, skates, and rays). In the present study the distribution of Zebrin II immunopositivity was examined in the Atlantic stingray and the Southern stingray. Western-blot analysis demonstrates that the Zebrin II antibody recognizes an antigen of the same molecular weight in stingrays, teleosts, and mammals. In stingrays, anti-Zebrin II immunohistochemistry reveals a staining pattern in which all Purkinje cells are immunopositive, no banding pattern or zonal compartmentation is observed. Purkinje cell axon projections to the cerebellar nucleus and the octaval nuclei are also revealed. Within the octaval nuclei, immunopositive Purkinje cell axon terminals and boutons en passant were found in the anterior, descending, and posterior nuclei. These immunopositive profiles are found throughout these nuclei, but they are most dense in the lateral and ventral portions. Except for the dorsolateralmost portion, the magnocellular nucleus does not appear to receive Purkinje cell inputs. Based on these results it is concluded that the Zebrin II distribution pattern in which all Purkinje cells of the cerebellar corpus are immunopositive is the primitive condition for jawed vertebrates. PMID- 9406647 TI - Specific detection of Coxsackie viruses A by the polymerase chain reaction. AB - BACKGROUND: Most of the Coxsackie virus A strains are difficult to identify using traditional diagnostic methods such as virus isolation followed by neutralization with type-specific antisera. For the laboratory diagnoses of infections with Coxsackie viruses A, inoculation into newborn mice has traditional been the method of choice. However, such investigations are complicated and time consuming. OBJECTIVES: To develop a reverse transcriptase (RT) and polymerase chain reaction (PCR) assay for specific detection of Coxsackie viruses A. STUDY DESIGN: A total of 43 clinical specimens containing Coxsackie viruses A, B or echoviruses were investigated retrospectively. Nineteen samples were Coxsackie virus A positive, whereas 24 samples were positive for Coxsackie viruses B or echoviruses. Thirteen non-typable specimens from eight patients were also included, since they were characterized as enterovirus-like by electron microscopy. RESULTS: All the specimens containing Coxsackie virus A were positive with the Coxsackie virus A PCR assay. In addition, five out of eight samples characterized as enterovirus-like by electron microscopy were PCR positive. The PCR assay did not amplify Coxsackie viruses B or echoviruses identified in our laboratory. CONCLUSION: The RT-PCR protocol established here should provide a useful alternative to the complicated and time-consuming diagnostic method based on live animals. PMID- 9406646 TI - Current aspects of diagnosis and treatment of cytomegalovirus infections in infants. AB - BACKGROUND: Human cytomegalovirus (CMV) is the most common cause of congenital and perinatal infections throughout the world. High prevalence of CMV infection is associated mainly with universal breast feeding practices rather than with crowding or poverty. Perinatal CMV infection usually follows a benign course in immunocompetent individuals, but the virus remains latent or persistent in the host cell thereafter. OBJECTIVE: As a clinical diagnosis of CMV infection is generally not easy, rapid and accurate laboratory diagnosis of CMV infection is required for appropriate patient management. Numerous anti-CMV compounds including ganciclovir, foscarnet and cidofovir have been reported, most of them are unlikely in a clinical trial for perinatal CMV infection in immunocompetent individuals. Understanding the epidemiology of CMV is a key element in the development of strategies for prevention and treatment of infection. STUDY DESIGN: This review focuses on recent advances in the diagnosis and treatment of perinatal CMV infections in infants. CONCLUSIONS: Entirely new approaches to prevention and treatment of CMV infections in infants are necessary, including antiviral interventions and the development of a vaccine strategy. PMID- 9406648 TI - The prevalence of hepatitis C virus types in patients of the same geographic area, according to the source of infection and liver disease. AB - BACKGROUND: The duration and stage of hepatitis C might be associated with the source of infection and hepatitis C virus (HCV) types. OBJECTIVE: We studied the relationship among the different HCV types, source, duration, and stage of infection in 100 patients from the Apulia, southern Italy, selected from consecutive clinical records. They were 20 parenterally infected haemophiliacs with 10-20 years of disease history, but without cirrhosis; 20 patients (matched for sex, age and disease) and without known risk factor for parenteral infections; 60 patients with community acquired infection (ten with CAH and ten with cirrhosis with less than 20 years disease history; 20 with cirrhosis and hepatocellular carcinoma (HCC) and more than 20 years of liver disease and 20 matched cases with cirrhosis without HCC). RESULTS: Type 1 and 2 HCVs had comparable prevalence in patients with long lasting and recent HCV infection, 56 and 64%, 26 and 30% respectively. HCV type 3 was found in 6.5-12% of the patients with recent HCV infection, but it was not detected in those with infection longer than 20 years. Type 1 b HCV was more frequently found in HCC patients (68% of cases) than in the other forms of liver disease. The opposite was observed for HCV types (2 and 3). CONCLUSIONS: The prevalence of the different HCV types appears associated with the source and duration of the infection. The interesting association between HCV type 1 b and HCC prompts further studies in larger series of patients. PMID- 9406649 TI - A new generation of serum anti-HIV antibody immunocapture assay for saliva testing. AB - BACKGROUND: Epidemiological studies need easier tools to evaluate HIV prevalence, particularly in high-risk groups under difficult field conditions. Testing saliva antibody with accurate immunoassay could serve as an alternative to serum testing. OBJECTIVES: To evaluate performances on saliva of a novel commercially available assay for anti-HIV antibody. STUDY DESIGN: Samples of saliva from 530 patients were tested for the presence of HIV antibodies with a third generation commercially available serum-screening kit and the use of a sample diluent adapted to saliva. RESULTS: Compared with serum sampling the sensitivity and specificity of oral sampling were respectively, 100 and 99.8%. CONCLUSION: The ICE HIV-1.0.2 assay used on saliva could be an efficient and non-invasive epidemiological tool for HIV testing. PMID- 9406650 TI - Biosensor technology and surface plasmon resonance for real-time detection of HIV 1 genomic sequences amplified by polymerase chain reaction. AB - BACKGROUND: The recent development of biosensor technologies for biospecific interaction analysis enables the monitoring of a variety of molecular reactions in real time by surface plasmon resonance (SPR). If the ligand is a biotinylated single stranded DNA, this technology could monitor DNA-DNA hybridization. This approach could be of great interest in virology, since the hybridization step is oftenly required to confirm specificity of molecular diagnosis. OBJECTIVES: To determine whether real-time molecular diagnosis of human immunodeficiency virus type I (HIV-1) could be performed using biosensors and SPR technology. STUDY DESIGN: Specific hybridization of a biotinylated HIV-1 oligonucleotide probe immobilized on a sensor chip to single stranded DNA obtained by asymmetric polymerase-chain reaction (PCR) was determined using the BIAcore biosensor. RESULTS: Direct injection of asymmetric PCR to a sensor chip carrying an internal HIV-1 oligonucleotide probe allows detection of hybridization by SPR using biosensor technology. This enabled us to apply a real-time, one-step, non radioactive protocol to demonstrate the specificity of amplification of HIV-1 genomic sequences by PCR. CONCLUSION: The procedure described in this study for HIV-1 detection is simple, fast (PCR and SPR analyses take 30 min), reproducible and could be proposed as an integral part of automated diagnostic systems based on the use of laboratory workstations and biosensors for DNA isolation, preparation of PCR reactions and analysis of PCR products. PMID- 9406651 TI - Molecular epidemiology of rabies epizootics in Texas. AB - BACKGROUND: Texas is in the midst of two independent epizootics of rabies, involving coyotes (Canis latrans) and domestic dogs (Canis familiaris) in southern Texas and grey foxes (Urocyon cinereoargenteus) in west central Texas. The domestic dog/coyote (DDC) and grey for (TF) rabies virus variants cannot be differentiated by antigenic typing with currently available monoclonal antibodies. These two variants also cannot be distinguished from a third variant, Sonora dog (SD) rabies, that is not enzootic in Texas, but occasionally occurs in animals along the western border with Mexico. OBJECTIVES: To determine a method for the differentiation of the DDC. TF and SD variants, which is essential for epidemiologic monitoring of the Oral Rabies Vaccination Program (ORVP), a program instituted to control rabies in coyotes and grey foxes in Texas. STUDY DESIGN: Primers complementary to nucleoprotein sequence of either the DDC or TF rabies virus permit specific reverse transcription and amplification by polymerase chain reaction. In addition, general primers, which recognize a broad range of rabies variants, used in conjunction with a restriction digest for the differentiation of DDC, TF of SD rabies virus were investigated. RESULTS AND CONCLUSIONS: Of 122 specimens tested with specific primers. 111 (91%) were specifically identified as either DDC (33 samples) or TF (78 samples). Overly stringent conditions, enzyme inhibitors, or limiting RNA may account for the 11 non-amplifications. Amplification of RNA under less stringent conditions, with primers recognizing a broad range of rabies variants followed by digestion with either restriction enzyme Desulfovibrio desulfuricans I (Dde I) or Haemophilus influenzae Rf. (HinfI), was used to identify the 11 isolates that did not amplify with specific primers (6 DDC, 4 TF and 1 SD). In addition to these 11 isolates, the less stringent method of amplification, followed by enzyme digestion has identified a total of 125 additional specimens (26 DDC, 94 TF and 5 SD) that were not tested by variant-specific amplification. These data provide a means to track the spread of the different rabies virus variants and allow the ORVP to plan its vaccine disbursement by defining the two epizootic boundaries. PMID- 9406652 TI - Secretory IgM and IgA antibodies to respiratory syncytial virus in nasopharyngeal aspirates: a diagnostic supplement to antigen detection. AB - BACKGROUND: RSV-shedding during an RSV-infection declines dramatically after the first week of infection. It could be of interest to be able to diagnose RSV infection for a longer period of time by detection of specific RSV-IgM and RSV IgA in nasopharyngeal aspirates (NPA) in order to minimize unnecessary antibiotics. OBJECTIVES: To evaluate an ELISA to detect specific RSV-IgM and RSV IgA in NPA as a supplement to RSV-antigen detection. STUDY DESIGN: A total of 104 NPA from 101 children (median age 9 months) with acute respiratory disease (group 1) admitted to hospital and consecutive NPA (collected on day 0, 7, 14, 30 and 60) from 11 children (median age 3 months) with a proven RSV infection (group 2) were collected. All NPA from group 1 were analysed for RSV-antigen, RSV-IgM and RSV-IgA. NPA from group 2 were analysed for RSV-IgM and RSV-IgA. RESULTS: Thirty five NPA in group 1 were positive for RSV-antigen and 64 were positive for RSV antigen test alone found 44% and the RSV-IgM test alone found 80%. In group 2 8/11 (73%) has an excellent RSV-IgM response day 7, the rest responded later. Only 5/11 (46%) had a less pronounced RSV-IgA response on day 7, three cases responded later and three did not respond at all. RSV-IgM disappeared in 8/11 (73%) and RSV-IgA in 7/8 (88%) between day 30-60. CONCLUSIONS: Specific RSV-IgM is a valuable supplement to RSV-antigen detection for the diagnosis of acute and recent RSV infection. PMID- 9406653 TI - Rapid detection of different RNA respiratory virus species by multiplex RT-PCR: application to clinical specimens. AB - BACKGROUND: The polymerase chain reaction (PCR) applied in diagnostic and epidemiologic investigations is very useful for sensitivity, specificity and time saving. OBJECTIVE: We have developed a method for the detection of genomic RNA of two different species of virus, the influenza A virus (IA) and the respiratory syncytial virus (RS), which are responsible for clinical similarities. We applied this multiplex RT-PCR protocol on clinical specimens. STUDY DESIGN: We describe a method which allows rapid diagnosis by performing a single retro-transcriptase (RT) reaction associated with the PCR (multiplex RT-PCR) on different genomes in a single sample. We have evaluated the sensitivity and the specificity of the multiplex test on positive controls, then, on RNA extracted from clinical specimens harvested from 15 children with respiratory symptoms during the spring winter season 1997. RESULTS AND CONCLUSIONS: The multiplex RT-PCR protocol, applied to respiratory specimens, allows the investigation of RNA IA virus and RS virus in a single sample at the same time. The detection of the etiologic viral agent is rapid and it is possible to evaluate incidental simultaneous infections. PMID- 9406654 TI - Measles virus-specific immunoglobulin G subclass response in serum and cerebrospinal fluid. AB - BACKGROUND: While many previous studies have focused on the impairment in the cellular immunity during measles virus infection, to date, a limited amount of data is available concerning the virus-specific IgG subclass response during measles virus infection. OBJECTIVE: The purpose of this study is to analyze the measles virus infection on the basis of virus-specific IgG subclass (G 1 and G 3). STUDY DESIGN: Frozen-stored, serum and/or cerebospinal fluid samples from three groups of patients were tested retrospectively; Group 1 comprised 14 patients with measles primary infection, group 2, ten patients with reinfection/vaccine failure, and group 3, seven patients with subacute sclerosing panencephalitis. The method used was a modified ELISA method utilizing the Enzygnost IgG detection kit with mouse-monoclonal antibodies (clone HP6091 for IgG 1 and clone HP6050 for IgG 3). Avidity testing for each subclass IgG was also performed for selected samples by means of an 8 M urea-denaturation method. RESULTS: In group 1, the IgG 3 could be detected in serum within 7 days from the onset of rash more frequently than IgG 1. In the cases of group 2, both subclasses were detected in very acute phase serum samples. In these cases, the IgG 1-specific avidity was always higher than that of IgG 3. In group 3, the subclass IgGs detected in the cerebrospinal fluid had a lower avidity than those in the serum. CONCLUSIONS: Our results suggested that in measles virus infection, like other viral infections, the IgG 3 response normally occurs before the IgG 1 response, and plays a major role in the acute phase immunity during the primary infection, while the IgG 1 plays a major role in the maintenance of immunity. Continuously produced IgG 1 and IgG 3 in the central nervous system in cases of subacute sclerosing panencephalitis may be derived from cell populations different from those in the blood. PMID- 9406655 TI - Structure and function of uridine diphosphate glucuronosyltransferases. AB - 1. The uridine diphosphate (UDP)-glucuronosyltransferases (UGT) are a family of enzymes that catalyse the covalent addition of glucuronic acid to a wide range of lipophilic chemicals. They play a major role in the detoxification of many exogenous and endogenous compounds by generating products that are more polar and, thus, more readily excreted in bile or urine. 2. Inherited deficiencies in UGT forms are deleterious, as exemplified by the debilitating effects of hyperbilirubinaemia and neurotoxicity in subjects with mutations in the enzyme that converts bilirubin to its more polar glucuronide. 3. The UGT protein can be conceptually divided into two domains with the amino-terminal half of the protein demonstrating greater sequence divergence between isoforms. This region apparently determines aglycone specificity. The aglycone binding site is presumed to be a 'loose' fit, as many structurally diverse substrates can be bound by the same UGT isoform. The carboxyl-terminal half, which is more conserved in sequence between different isoforms, is believed to contain a binding site for the cosubstrate UDP glucuronic acid (UDPGA). 4. Uridine diphosphate glucuronosyltransferase is localized to the endoplasmic reticulum (ER) and spans the membrane with a type I topology. The putative transmembrane domain is located near the carboxyl terminus of the protein such that only a small portion of the protein resides in the cytosol. This cytosolic tail is believed to contain an ER targeting signal. The major portion of the protein is located in the ER lumen, including the proposed substrate-binding domains and the catalytic site. 5. The microsomal membrane impedes the access of UDPGA to the active site, resulting in latency of UGT activity in intact ER-derived microsomes. Active transport of UDPGA is believed to occur in hepatocytes, but the transport system has not been fully characterized. Uridine diphosphate glucuronosyltransferase activity is also highly lipid dependent and the enzyme may contain regions of membrane association in addition to the transmembrane domain. PMID- 9406656 TI - Mitochondrial function in rat renal cortex in response to proteinuria and iron. AB - 1. Proximal tubular cell dysfunction in chronic glomerular disease (CGD) has been ascribed, in part, to reabsorption of transferrin-iron from tubular fluid and subsequent cytosolic peroxidative injury. To investigate a possible role for altered mitochondrial function in tubular cell injury in CGD, renal cortical mitochondrial respiratory function was examined in rats with adriamycin nephrosis. 2. State 4 (resting) respiration was increased in adriamycin nephrosis in comparison with control (51 +/- 2 vs 43 +/- 2 ng atoms oxygen (O)/min per mg protein, respectively; P < 0.02). 3. Mitochondrial iron concentration was increased in nephrotic rats compared with control (9.52 +/- 0.70 vs 5.97 +/- 0.26 nmol Fe/mg protein, respectively; P < 0.001) and rates of state 3, state 4 and uncoupled respiration and the severity of proteinuria correlated with mitochondrial iron concentration. 4. To further define the relationship between mitochondrial iron accumulation and altered respiratory function, rats were loaded with iron. 5. In comparison with control, acute iron loading of normal rats impaired creatinine clearance (1.48 +/- 0.02 vs 0.40 +/- 0.29 mL/min), increased kidney weight (1.33 +/- 0.07 vs 1.74 +/- 0.14 g) and impaired mitochondrial enzyme activity (e.g. cytochrome oxidase 185.0 +/- 46.6 vs 362.0 +/ 32.8 delta log [cytochrome C]/min per mg protein; P < 0.05), but had no significant effect on rates of mitochondrial respiration or on mitochondrial fragility. 6. Mitochondrial iron concentration was not increased by iron loading, despite a similar increment in cytoplasmic iron to that seen in rats with adriamycin nephrosis. 7. In summary, resting mitochondrial respiration is increased in nephrotic rats in proportion to mitochondrial iron accumulation. Changes in mitochondrial oxygen consumption do not appear to be a primary event in the tubular cell injury of iron loading. PMID- 9406657 TI - Evidence for direct interaction of ketamine with alpha 1- and beta 2 adrenoceptors. AB - 1. Ketamine has a number of effects that suggest that it may interact with alpha- and beta-adrenoceptors. To date, the experimental evidence for this has been indirect and has been based on physiological studies using competitive blocking agents. In the present study we sought to determine from receptor binding studies whether ketamine binds directly to alpha- and beta-adrenoceptors. 2. Membrane preparations of alpha 1- and beta 2-adrenergic binding sites were obtained from urinary bladder and urethrae of sheep. These binding sites were characterized by saturation analyses using [3H]-prazosin for alpha 1-adrenoceptor binding sites and [125I]-cyanopindolol (CYP) for the beta 2-adrenoceptor binding sites. The receptors were further characterized by displacement studies using selective and non-selective antagonists. 3. Studies in which ketamine was used to displace [3H] prazosin revealed a Kd of 3.40 +/- 1.23 x 10(-3) mol/L for ketamine binding to alpha 1-adrenoceptors. Displacement studies of [125I]-CYP by ketamine showed a Kd of 0.35 +/- 0.03 x 10(-3) mol/L for ketamine binding to beta 2-adrenoceptors. 4. We conclude that ketamine interacts directly with both alpha 1- and beta 2 adrenoceptors and that such interactions probably explain the reported effects of this agent on the vasculature and the bronchial tree. PMID- 9406658 TI - Potentiation of dipsogenic actions by centrally administered type-C natriuretic peptide in spontaneously hypertensive but not Wistar-Kyoto rats. AB - 1. The effects of type-C natriuretic polypeptides (CNP) on the central dipsogenic and pressor responses to angiotensin II (AngII) were studied by the administration of agents into the lateral cerebral ventricle under conscious and unrestrained conditions in normotensive Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR). 2. The fluid intake induced by AngII (25 ng) and water deprivation were potentiated after pretreatment with CNP in SHR but not in WKY rats. However, carbachol-induced water intake was not altered by pretreatment with CNP (2.5 micrograms) in either WKY rats or SHR. 3. In contrast, CNP did not influence the pressor responses to AngII in either WKY rats or SHR. PMID- 9406659 TI - Volume-activated taurine transport is differentially activated in human cervical cancer HT-3 cells but not in human papillomavirus-immortalized Z183A and normal cervical epithelial cells. AB - 1. Previous studies demonstrate that volume-sensitive chloride currents are distinctly activated in cervical cancer cells, but not in human papillomavirus (HPV)-immortalized and normal cervical cells. In the present study, the Na(+) independent volume-activated transport of taurine in three cervical cell types was investigated. 2. Osmotic swelling of cervical cancer HT-3 cells suspended in Na(+)-free hypotonic medium led to increased membrane uptake of taurine. This taurine uptake was effectively blocked by various Cl- channel blockers with a similar potency in blocking volume-sensitive Cl- channels: 1,9-dideoxyforskolin > 5-nitro-2-(3-phenyl-propylamino)-benzoic acid (NPPB) > 4-acetamido-4' isothiocyanastilbene-2,2'-disulphonic acid (SITS) > 4,4'-diisothio cyanatostilbene-2,2-disulphonic acid (DIDS) > furosemide. The taurine influx was also abolished by pertussis toxin. In contrast, Na(+)-independent volume activated taurine transport was not significantly activated in HPV-immortalized Z183A cells and in normal cervical cells. 3. Exposure of HT-3 cells to hypotonic medium also resulted in a marked increase in taurine efflux. The volume-activated taurine efflux was osmolarity dependent and the pattern of pharmacological inhibition by Cl- channel blockers was indistinguishable from that for taurine uptake. 4. These results suggest that volume-sensitive Cl- channels in HT-3 cells can mediate the transport of amino acids. In addition, the pertussis toxin sensitive G-protein is linked with the activation of this transport mechanism. PMID- 9406660 TI - Autonomic control of bronchial circulation in awake sheep during rest and behaviour. AB - 1. We tested the hypothesis that the pattern and the intensity of autonomic mechanisms causing vasoconstriction in the resting bronchial circulation of awake dogs also exists in awake sheep. It was also postulated that sighing behaviour and the associated bronchovascular dilatation induced by non-adrenergic, non cholinergic (NANC) mechanisms observed in the dog exist in sheep. 2. Bronchial arterial blood flow to lower airways of both lungs of awake sheep was measured continuously using pulsed Doppler flow probes mounted on the bronchial artery at prior thoracotomy. 3. Cumulative and factorial analysis of responses to randomized combinations of autonomic alpha 1-, alpha 2-, beta 1- and beta 2 adrenoceptors and cholinoceptor autonomic blockade suggests that resting vasoconstrictor activity is less in sheep than in dogs. At normal aortic pressure, the autonomic activity of these receptor groups in the sheep lowers bronchial blood flow and conductance by 30%, whereas in the awake dog, the corresponding autonomic effect is 50%. 4. Tonic autonomic control of bronchial conductance can be partitioned in sheep to show significant and separate alpha- and beta-adrenoceptor vasoconstrictor activity at a ratio of 1.8:1, an effect normally offset by a weaker vasodilator alpha-/beta-adrenoceptor interaction. In contrast to the situation in awake dogs, cholinoceptors do not play a role in awake sheep. 5. Nitric oxide (NO) synthase inhibition in sheep using NG-nitro-L arginine following blockade of alpha- and beta-adrenoceptors and cholinoceptors causes hypertension, but minor changes, if any, in pulmonary pressures or heart rate. Bronchial flow and conductance, however, fall from a higher resting conductance by approximately 50%, suggesting that, normally, resting bronchial flow conductance is dominated by strong tonic NO vasodilator effects that interact with weaker tonic autonomic vasoconstrictor effects. 6. Superimposed (respiratory) behaviours of sighing, sneezing and coughing, which involve negative swings in intrathoracic pressure and the movement of inspired air, evoke large active bronchovascular dilator effects. These appear to be largely NANC in origin and appear to be dependent, in part, on mechanisms associated with NO release. It is postulated that the C-fibre axon reflex using substance P, calcitonin gene-related peptide and neurokinin A may be involved. Vocalization and eructation do not evoke bronchovascular effects. PMID- 9406661 TI - Rapid reversal of endothelial dysfunction in hypercholesterolaemic rabbits treated with simvastatin and pravastatin. AB - 1. The main objective of the present study was to verify the speed with which two 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors, simvastatin and pravastatin, could revert endothelial cell dysfunction in hypercholesterolaemic rabbits. An attempt was also made to correlate the plasma cholesterol level and the tissue cholesterol and malondialdehyde (MDA) contents of the aortae with the endothelium-dependent relaxation on the assumption that any endothelial dysfunction could be rapidly and partially reversed, even in the presence of relatively high serum cholesterol levels. 2. Ninety-one male New Zealand white rabbits were randomly assigned to hypercholesterolaemic (control), simvastatin or pravastatin groups. All rabbits were fed a diet supplemented with cholesterol (0.5%) and coconut oil (2%) for 8 weeks. Simvastatin (10 and 20 mg/day) and pravastatin (15 and 30 mg/day) were administered 6, 4 and 2 days before the end of the experiment. At the end of the 8th week, animals were killed and the aortae were removed for histological examination as well as for the measurement of cholesterol and MDA contents and for endothelium-dependent relaxation studies. 3. The results showed that significant improvement in endothelium-dependent relaxation was obtained only with pravastatin and only with 4 or 6 days of administration. In these cases, the cholesterol and MDA contents of the vessel wall were reduced, although no significant changes were observed in plasma total cholesterol. Higher doses of the drugs did not alter these results. 4. We conclude that pravastatin enhances endothelium-dependent relaxation when administered to cholesterol-fed rabbits, probably via an anti-oxidant action. This effect, which was observed to start on the 4th day of drug administration, may represent a new therapeutic approach for the treatment of acute coronary syndromes in hypercholesterolaemic patients. PMID- 9406662 TI - Coronary endothelial function in open heart surgery. AB - 1. During open heart surgery, the heart is arrested and protected by hyperkalaemic cardioplegia. The coronary endothelium may be damaged by ischaemia reperfusion and cardioplegia. Subsequently, this may affect cardiac function immediately after cardiac surgery and cause mortality or morbidity. 2. Our studies have investigated coronary endothelial function after exposure to hyperkalaemia (K+ 20 or 50 mmol/L). Endothelium-dependent relaxation and hyperpolarization of the coronary smooth muscle and intracellular free calcium concentration in the endothelial cell were measured with regard to K+ exposure. 3. Endothelium-derived hyperpolarizing factor (EDHF)-mediated relaxation to A23187, bradykinin, and substance P in the presence of either U46619 (10 nmol/L)- or K+ (25 mmol/L)-induced contraction was reduced after exposure to either 20 or 50 mmol/L K+. 4. The hyperpolarization of the membrane potential in response to the endothelium-derived relaxing factor (EDRF) stimuli was also reduced by exposure to K+. 5. The intracellular free calcium concentration remained unchanged after exposure to hyperkalaemia. 6. We conclude that the EDHF-mediated coronary endothelial function is impaired after exposure to hyperkalaemic cardioplegia. The impairment of this function is due to the changed effect of EDHF on the smooth muscle cell, probably through partially depolarizing the membrane and affecting K+ channels rather than alteration of its biosynthesis/release in the endothelial cell. It may be of use to search for a new cardioplegia that preserves this endothelial function during open heart surgery. PMID- 9406663 TI - Extracts of Ginkgo biloba and ginsenosides exert cerebral vasorelaxation via a nitric oxide pathway. AB - 1. Extracts from the leaves of Ginkgo biloba (EGb) and ginsenosides (GS) have been reported to be effective at increasing vascular relaxation. In the present study, the actions of EGb and GS on the vascular functions of porcine basilar arteries were investigated in vitro using tissue bath techniques. 2. Both EGb and GS relaxed the basilar artery in a concentration-dependent and partly endothelium dependent manner. However, EGb appeared to be more potent than GS. Relaxation induced by transmural nerve stimulation (TNS) was significantly enhanced by EGb (7.5, 15 and 30 micrograms/mL) and GS (20, 40 and 80 micrograms/mL) in both endothelium-intact and -denuded basilar arteries. Enhanced TNS-induced relaxations were abolished by 0.3 mmol/L N-L-arginine. 3. The present study demonstrates that nitric oxide plays a primary role in TNS-induced relaxation as well as in EGb- and GS-enhanced relaxation within the cerebral vasculature. In addition, our data support the potential of these compounds as therapeutic strategies in cerebral ischaemia and other related vascular dysfunctions. PMID- 9406664 TI - Management of branch pulmonary artery stenosis: balloon angioplasty or endovascular stenting. AB - 1. The surgical outcome of congenital heart diseases may be adversely affected by residual branch pulmonary artery stenosis, which is difficult to treat surgically. 2. The objective of the present study was to evaluate the effectiveness, safety and follow-up results of two transcatheter procedures, balloon angioplasty and endovascular stenting, for treatment of branch pulmonary artery stenosis. 3. From December 1988 to March 1997, 22 children (group 1) underwent 30 balloon angioplasties and 12 children (group 2) underwent 14 endovascular stent implantations. The overall success rates for groups 1 and 2 were 67 (20/30) and 93% (13/14), respectively, with significant increases in vessel diameter (P < 0.001, t-test, 29 d.f.; P = 0.0001, t-test, 12 d.f., respectively) and decreases in pressure gradient (P < 0.0001, t-test, 29 d.f.; P = 0.001; t-test, 12 d.f., respectively). One death (3.3%) in group 1 was due to post-dilation reactive pulmonary hypertension. No mortality occurred in group 2 children, but two migrated stents required re-implantation of another stent. Mean follow-up periods were 28.2 and 26.3 months for groups 1 and 2, respectively. For group 1, no significant change in vessel diameter or pressure gradient was noted on recatheterization in 12 patients. Restenosis occurred in four of 16 (25%) initial successes. Balloon redilation in four vessels were all successful. For group 2 children, although vessel diameter remained unchanged, a significant increase in pressure gradient (P = 0.02; t-test, 11 d.f.) was noted on recatheterization. Balloon dilations on two narrowed stents caused by intimal proliferation showed only partial improvements. 4. In conclusion, both balloon angioplasty and endovascular stent implantation are effective and safe, with satisfactory intermediate-term results, for the treatment of branch pulmonary artery stenosis. Balloon angioplasty is the choice for initial treatment, whereas stent implantation, the long-term outcome of which remains to be determined, should be reserved for older children after repeated failures with balloon dilation. PMID- 9406665 TI - Local hypoxia does not elevate adenosine output from isolated gracilis muscle in anaesthetized dogs. AB - 1. The influence of local hypoxia on adenosine and lactate output from isolated perfused gracilis muscle was studied in anaesthetized dogs. 2. Oxygen tension in the arterial blood supplying the muscle was reduced by a membrane lung from 145.9 +/- 28.9 to 52.9 +/- 2.6 (moderate hypoxia) or 30.0 +/- 1.2 mmHg (severe hypoxia). 3. Moderate hypoxia did not significantly alter vascular resistance, but severe hypoxia reduced arterial perfusion pressure from 199.0 +/- 13.6 to 122.6 +/- 8.7 mmHg. 4. Veno-arterial (V-A) lactate was 0.47 +/- 0.13 mmol/L in normoxia; neither level of hypoxia changed it significantly. Veno-arterial adenosine was 74 +/- 78 nmol/L in normoxia. Moderate hypoxia decreased this to 36 +/- 59 nmol/L (P < 0.05), but the level of V-A adenosine in severe hypoxia (52 +/- 96 nmol/L) was similar to that in normoxia. 5. These data confirm that hypoxia does not directly stimulate adenosine output from oxidative skeletal muscle. PMID- 9406666 TI - Plasma concentration of brain natriuretic peptide is related to diastolic function in hypertension. AB - 1. The plasma brain natriuretic peptide (BNP) concentration is elevated in patients with essential hypertension and normal systolic function. This may be related to left ventricular hypertrophy or diastolic dysfunction, both of which commonly occur in hypertension. 2. Echocardiography was performed on 32 patients with newly diagnosed untreated mild-to-moderate hypertension (19 men, 13 women; mean +/- SD age 51 +/- 15 years; diastolic blood pressure 99 +/- 12 mmHg; systolic blood pressure 153.2 +/- 18.0 mmHg; plasma creatinine 86 +/- 15 mumol/L; creatinine clearance 92.2 +/- 20.5 mL/min; left ventricular mass index 116 +/- 28 g/m2; left ventricular ejection fraction 66 +/- 9%). A 15 mL peripheral venous blood sample was obtained at the time of echocardiography for radioimmunoassay of BNP. 3. Sixteen patients had abnormal Doppler transmittal flow (E/A ratio < 1) and a higher median plasma BNP concentration compared with those patients with E/A > or = 1 (12.9 vs 5.9 pmol/L, respectively; P = 0.006). The plasma BNP level correlated significantly with E/A ratio (r = -0.50; P = 0.035). Multivariate analysis showed that the E/A ratio is related to plasma BNP, independent of age and blood pressure. 4. Our results suggest that the plasma BNP level is influenced by diastolic dysfunction. Further studies are needed to determine whether assay of plasma BNP helps to identify patients with diastolic dysfunction. PMID- 9406667 TI - Prostanoid action on the human pulmonary vascular system. AB - 1. Four types of prostanoid receptor are present on pulmonary arterial vessels of man. Thromboxane (TP) receptors mediate constriction and are blocked by antagonists such as BAY u-3405, GR 32,191 and EP 169. Prostaglandin (PG) EP3 receptors also mediate constriction, the agonist potency ranking being SC 46,275 > sulprostone > misoprostol > or = PGE2; this action needs to be borne in mind when PGE analogues are used therapeutically. 2. Prostaglandin E2 causes relaxation in a few pulmonary artery preparations: an EP2 receptor may be involved. Prostacyclin, acting through i.p. receptors, consistently produces relaxation and studies are in progress to determine the contribution made by K(+) channel opening. Agonist potencies of stable prostacyclin analogues and non prostanoid prostacyclin mimetics, such as BMY 45,778 and the novel diphenylindole CU 23, on human pulmonary artery and platelets are well correlated. Interestingly, the non-prostanoid mimetics show persistent relaxant effects in vitro, which may be related to their high lipophilicities. 3. Prostacyclin and iloprost are being used to treat severe pulmonary hypertension; further study of the pharmacodynamic and pharmacokinetic properties of other i.p. receptor agonists could produce improved therapy. PMID- 9406668 TI - Role of lipoprotein-X in foam cell formation and rat mesangial cell proliferation. AB - 1. In the present study, the effect of reconstituted lipoprotein-X (rLp-X) on lipid accumulation and foam cell formation in rat peritoneal macrophages was evaluated. Furthermore, the combined effect of rLp-X and macrophages on mesangial cell proliferation was examined. 2. Incubation of macrophages with rLp-X (177 and 387 nmol unesterified cholesterol (FC)/mL) resulted in an increase of cellular cholesterol (162%) and cholesteryl esters (223 to 245%) relative to control. 3. Oil Red O staining of macrophages treated with rLp-X revealed the presence of foam cells. 4. In conclusion, rLp-X had no effect on the proliferation of mesangial cells incubated in macrophage-conditioned medium. PMID- 9406669 TI - Reduced inhibitory actions of adenosine A1 and kappa 1-opioid receptor agonists on beta-adrenoceptors in spontaneously hypertensive rat heart. AB - 1. The modulatory actions of both adenosine A1 and kappa 1-opioid receptor agonists on beta-adrenoceptor stimulation in the heart of both spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto (WKY) rats were compared. 2. In both types of rats, both R(-)-N6-(2-phenylisopropyl)adenosine (R-PIA), an adenosine A1 receptor agonist, and U50 488H, a kappa 1-opioid receptor agonist, inhibited the stimulatory effects of beta-adrenoceptor activation on electrically induced [Ca2+]i transients measured by a spectrofluorometric method with fura 2/AM as the calcium indicator. The effects of these two agonists were blocked by their respective antagonists, namely 8-cyclopentyl-1,3-diprolxanthine and norbinaltorphimine. 3. The inhibitory actions of both R-PIA and U50 488H on beta adrenoceptor augmentation of electrically induced [Ca2+]i transients in the heart were more significantly reduced in SHR than in WKY rats, suggesting the negative modulatory actions of endogenous substances on beta-adrenoceptors were impaired in SHR, which may contribute to hypertension. PMID- 9406670 TI - New insights into the treatment of hypertension. AB - 1. The choice of initial pharmacological therapy is one area where the different hypertension management guidelines vary in recommending either the use of diuretics and beta-blockers as preferred drugs or choosing from any of the five major classes of antihypertensives. 2. Improvement in cardiovascular morbidity and mortality may have been shown most conclusively with diuretics and beta blockers, but the effects on coronary events with these drugs were less than predicted and the other agents have a number of theoretical advantages. 3. Recent worries regarding the possible adverse effects of calcium antagonists have led to a reappraisal of the risks and benefits of these drugs and antihypertensives in general. 4. Many patients, particularly the elderly, have other conditions that influence the choice of first-line therapy and ethnic variations in the effects of hypertension or the efficacy or side effects of drugs should also be taken into account. 5. There is considerable heterogeneity within the major categories of antihypertensive drugs, so it is important to distinguish the different subgroups, dosages and formulations that may be used. PMID- 9406671 TI - Electrical remodelling of chronic atrial fibrillation. AB - 1. It is now recognized that atrial fibrillation (AF) is not a benign condition, as it is associated with a 40% increase in mortality and a doubling of the risk of stroke. 2. The development of AF leads to mechanical, electrophysiological and cellular changes in the atria that tend to sustain AF. This process is known as atrial remodelling. 3. The three electrophysiological elements in the atria that initiate and sustain AF are: (i) shortening of the refractory period and an increase in dispersion; (ii) slowing of conduction velocity; and (iii) the presence of triggering foci. 4. As AF is a heterogeneous disorder, therapeutic strategies include the use of devices (pacemakers and atrial defibrillators), radiofrequency ablation (focal ablation or the creation of linear lines) and drug therapy that may reverse a remodelled atrium. PMID- 9406672 TI - Endothelial dysfunction exacerbates the impairment of relaxation by lysophosphatidylcholine in porcine coronary artery. AB - 1. Current evidence suggests that lysophosphatidylcholine (LPC), a component found in oxidized low-density lipoprotein (Ox-LDL), inhibits endothelium dependent relaxation (EDR) mediated by endothelium-derived relaxing factor (EDRF) and endothelium-derived hyperpolarizing factor (EDHF). An objective of the present study was to characterize the roles of the different elements of EDR in LPC-induced impairment within the porcine coronary artery. Concomitantly, we sought to determine whether impairment of one component of EDR would increase the sensitivity of the endothelium to LPC. 2. Bradykinin (0.1 nmol/L -0.3 mumol/L) relaxed U46,619 (30 nmol/L)-precontracted porcine coronary artery rings in a concentration-dependent manner. A reduction in the bradykinin-elicited response was observed in NG-nitro-L-arginine methyl ester (L-NAME; 300 mumol/L)- and ouabain (50 mumol/L)-treated rings. Pretreatment with LPC (20 mumol/L), which on its own had no effect on normal endothelial relaxation, resulted in further inhibition of EDRF- and EDHF-induced relaxations. 3. Our results demonstrate that EDRF and EDHF are the primary mediators of EDR in the porcine coronary artery. Our data also show that while a low concentration of LPC (20 mumol/L) does not impair EDR, it can evoke vascular dysfunction following blockade of either the effects of EDRF or EDHF. Therefore, these data suggest that the partially damaged vascular endothelium could be more sensitive to threshold levels of this atherogenic phospholipid. PMID- 9406673 TI - Epidemiology of cardiovascular risk factors in Hong Kong. AB - 1. To facilitate the development of appropriate population-wide coronary heart disease prevention strategies and to monitor their long-term impact, a comprehensive baseline Hong Kong Cardiovascular Risk Factor Prevalence Study was conducted in 1995-96. 2. The data obtained from 2875 men and women aged 25-74 years showed a high prevalence of diabetes mellitus and impaired glucose tolerance, particularly in those aged 65 years and over. Other risk factors, such as hypercholesterolaemia and hypertension as well as being overweight, were particularly prominent in older women. PMID- 9406674 TI - Endothelium-smooth muscle interactions in blood vessels. AB - 1. Blood vessel tone is determined both by smooth muscle and endothelial functions. In coronary arteries taken from rat (Fisher-Lewis) cardiac transplanted hearts, the inducible form of NOS (iNOS) in smooth muscle is more active, while acetylcholine-induced nitric oxide production in the endothelium is greatly diminished. This causes a greatly reduced myogenic constriction, in pressurized septal arteries taken from immunologically challenged transplanted hearts. 2. The sarcoplasmic reticulum (SR) of smooth muscle and the endoplasmic reticulum (ER) of endothelial cells sequester Ca2+ from the cytoplasm. This reduces the intracellular concentration of free Ca2+, which is necessary for the activation of cellular processes. The release of Ca2+ from internal stores occurs through ryanodine and IP3 recoptors located on the SR membrane. 3. The superficial SR/ER also interacts with ion exchangers and pumps in the plasma membrane. This allows for the superficial SR/ER to function in Ca2+ extrusion; for example, inhibition of the SR/ER Ca(2+)-ATPase (SERCA) partially inhibits the rate of loss Ca2+ from the cell. Recent data suggest that the SR Ca(2+)-ATPase and the Na(+)-Ca2+ exchanger of smooth muscle cells function in series; that is, Ca2+ uptake by the SR followed by release towards the exchanger to mediate extrusion. This interaction between the SERCA of the superficial SR and ion exchangers and pumps creates intracellular Ca2+ gradients. 4. The SERCA of the superficial, peripherally distributed SR/ER also serves to regulate Ca2+ entry from the extracellular space. This occurs in part by inhibition of the superficial buffer barrier function of the SR as well as by depletion of stimulated Ca2+ entry. 5. Ca2+ entry is also regulated in endothelial and smooth muscle cells by the membrane potential. Membrane hyperpolarization increases the driving force for Ca2+ entry into endothelial cells, which lack voltage-gated Ca2+ channels, and reduces open state probability of voltage-gated Ca2+ channels in vascular smooth muscle cells. The two cell types have electrical contact and interact in a dynamic manner to regulate blood vessel diameter. PMID- 9406675 TI - Passive smoking and coronary heart disease: a brief review. AB - 1. Whether passive smoking or environmental tobacco smoke (ETS) can cause coronary heart disease (CHD) is controversial. We have undertaken a comprehensive review of the epidemiological studies regarding the relationship between ETS and CHD. 2. We searched for all original papers and reviews and calculated pooled odds ratio using the Mantel-Haenszel method. 3. Ten prospective studies, nine case-control studies and one cross-sectional study were found. Almost all studies showed positive associations between ETS and CHD and a few were statistically significant, with dose-response relationships being evident. Six review papers calculated pooled estimates of the relative risks, which ranged from 1.23 to 1.51. Other studies also showed that ETS could have other cardiovascular effects. 4. The association observed is likely to be real. The criteria for causation of time sequence, consistency, coherence and biological plausibility are satisfied. We conclude that ETS may cause CHD, particularly in women who have never smoked. PMID- 9406676 TI - Myocarditis as systemic disease: new perspectives on pathogenesis. AB - 1. Myocarditis may be an early indicator of or may subsequently lead to dilated cardiomyopathy in humans. This hypothesis has evolved from research on viruses that induce myocarditis, wherein the coxsackie B group viruses (CVB) in the family Picornaviridae are the most common known viral infectants of heart muscle. 2. Many competing hypotheses exist as to the pathogenesis of CVB3-induced myocarditis, including direct virus-induced myocyte damage and immunopathological disease with autoimmune sequelae. Evidence to support the direct-damage and viral RNA-persistence hypothesis is derived from in situ hybridization and gene amplification studies. 3. Recent use of terminal deoxynucleotidyl transferase mediated nick-end labelling indicates that this injury in target organs is largely non-apoptotic in nature. Most apoptotic bodies in cardiac tissue are derived from immune cells. 4. Beyond infection of heart muscle, CVB3 can also associate with, infect and persist in cells of immune origin. The CVB3 localizes to follicles in spleens and lymph nodes of the murine host and this particular localization may continue in mice susceptible to more aggressive myocarditis. Whether virus-immune cell association in these compartments is advantageous (or essential) to the host in the evolution of anti-viral immune responses or whether it is more advantageous to the virus in immunosuppression of the host is not known. 5. We suggest that CVB3 can directly perturb or alter the immune response, thereby delaying viral clearance from vulnerable systemic organs. Both host and viral genetic factors can influence susceptibility, persistence and disease progression. 6. Picornaviruses use a unique method for the initiation of translation, involving the internal binding of the ribosome on a sequence element of the 5' untranslated region, termed an internal ribosome entry site (IRES). 7. The IRES of CVB3 is located at approximately stem loops G, H and I, spanning nucloetides 530 and 630. Arrest of host translation is also a feature of picornavirus infection. Such regulation of host cell translation machinery no doubt fosters viral replication at the expense of the host cell. 8. Differences between cell types in the mechanisms, along with those at other key steps in the viral life cycle and in signalling via kinase pathways, may determine viral tropism and cellular destruction and the physiological outcome of neighbouring cells. PMID- 9406678 TI - Surgical treatment of endocarditis. AB - Since early investigators first suggested that the treatment of endocarditis should include valve replacement for infections not readily controlled with medical therapy alone, the role of surgery has become expanded, yet refined, to improve the outcome of patients with this potentially fatal disease. Innovative surgical techniques have also been developed in an effort to improve the results of surgical treatment for complex sequelae of invasive infections. This article examines the current indications for surgical intervention, compares the various surgical options, and assesses the expected short-and long-term outcome after valve replacement for patients with native valve or prosthetic valve endocarditis. PMID- 9406679 TI - Carnitine and its derivatives in cardiovascular disease. AB - Carnitine and its derivative propionyl-L-carnitine are endogenous cofactors which enhance carbohydrate metabolism and reduce the intracellular buildup of toxic metabolites in ischemic conditions. The carnitines have been, and are being used in a spectrum of diseases including multiple cardiovascular conditions. These include angina, acute myocardial infarction, postmyocardial infarction, congestive heart failure, peripheral vascular disease, dyslipidemia, and diabetes. Most published data on carnitine, propionyl-L-carnitine, and other carnitine congeners are favorable but the clinical trials have been relatively small. In currently used doses, these substances are virtually devoid of significant side effects. PMID- 9406677 TI - Thrombosis, antithrombotic agents, and the antithrombotic approach in cardiac disease. AB - To develop a rational approach to antithrombotic therapy, in cardiac disease, a sound understanding is required (1) of the hemostatic processes leading to thrombosis, (2) of the various antithrombotic agents, and (3) of the relative risks of thrombosis and thromboembolism in the various cardiac disease entities. With the understanding of pathogenesis and risk of thrombus formation, a rational approach to the use of antiplatelet and anticoagulant agents can be formulated. Those at high risk of thrombus formation should generally receive a high degree of antithrombotics and, depending on the pathophysiology of the thrombus, may benefit from the concomitant use of antiplatelet and anticoagulant agents. Those with a medium risk of thrombus formation may benefit with the use of an antiplatelet agent alone or anticoagulants alone. Patients at low risk of thrombus formation should not receive antithrombotics. Such rational approach to antithrombotic therapy serves as the basis of this article. PMID- 9406680 TI - Proceedings of the Sixth Conference on Radioimmunodetection and Radioimmunotherapy of Cancer. Princeton, New Jersey, October 10-12, 1996. PMID- 9406681 TI - Tribute to Ralph A. Reisfeld, Ph.D. PMID- 9406682 TI - Early results in the irrational design of new bifunctional chelators. AB - BACKGROUND: The development of a simple route for the synthesis of the N hydroxysuccinimide (NHS) ester of S-acetyl-protected mercaptoacetyltriglycine (MAG3) has opened the possibility of preparing novel bifunctional N3S chelators for technetium-99m (99mTc) and other radionuclides. In particular, the synthesis may be applied to a vast number of tripeptides in place of triglycine, to provide a "library" of bifunctional N3S chelators, each with unique properties related to the particular amino acid residues within each tripeptide. METHODS: The authors have synthesized by this simple route the NHS esters of four N3S chelators by reacting NHS-S-acetylthioglycolic acid with ala-gly-gly, phe-gly-gly, pro-gly gly, and ser-ser-ser, in addition to gly-gly-gly. Each bifunctional chelator was conjugated to biocytin as a model primary amine and radiolabeled with 99mTc. The properties of the four chelators were compared with MAG3 with respect to the stability of the label in saline and serum, the extent of serum protein binding, and the instability to cysteine challenge. RESULTS: A range of values was observed. Labeled mercaptoacetyltriserine showed stability towards transchelation to cysteine similar to that of MAG3 as well as lower serum protein binding; labeled mercaptoacetylalanyldiglycine showed slightly higher serum protein binding than labeled MAG3 but greater stability to cysteine challenge. CONCLUSIONS: The authors concluded that this simple synthesis and evaluation scheme may be used to prepare and screen a large library of bifunctional chelators for those with useful properties. PMID- 9406683 TI - Synthesis of bombesin analogues for radiolabeling with rhenium-188. AB - BACKGROUND: Gastrin-releasing peptide receptors (GRPR) are overexpressed in small cell lung carcinoma and some other human cancers. Small molecule peptides with antagonistic activities toward these receptors are potential radiotherapeutic agents. METHODS: A 7-amino acid analogue of bombesin (BBN) was synthesized through solid-phase techniques. The peptide was conjugated to trisuccin prior to cleavage from the resin. The conjugate was hydrogenated to remove the hydroxamate protecting benzyl groups followed by purification through reversed-phase high performance liquid chromatography (RP-HPLC). Rhenium-188 (188Re)-labeling of the trisuccin-peptide conjugate was performed by a SnCl2-reduced radioisotope and the labeled product was purified by RP-HPLC. The labeled conjugate was incubated with BNR-11 (3T3 mouse fibroblast cells stably transfected with murine GRPR) and PC-3 human prostate carcinoma GRPR positive cells. The nonradioactive peptide analogue was used as a competitive inhibitor and 125I-[Tyr4]-BBN was used as a positive control. RESULTS: Solid-phase and solution phase synthesis afforded the conjugates of the hydroxamate ligand trisuccin with the 7-amino acid BBN analogue. The molecules differed by either a direct attachment of the trisuccin to the peptide (TrisBBN) or connection through a 6-carbon linker (TrisC6BBN). The overall yield for each synthesis was approximately 20%. Both conjugates showed the correct molecular weights on mass spectroscopy. Radiolabeling of the conjugates with 188Re were performed in > or = 90% yield. Cell-binding assays performed with BNR-11 (TrisBBN and TrisC6BBN) and PC-3 (TrisBBN) cell lines resulted in positive binding. CONCLUSIONS: The synthesis and radiolabeling of Tris-BBN conjugates with 188Re were shown to be feasible. The yields of chemical syntheses and radiolabeling and positive binding of the radiolabeled conjugates to GRPR-positive tumor cells reveal promise in the use of these molecules for cancer imaging and therapy. More work is needed and is in progress to optimize the cell-binding properties. PMID- 9406684 TI - High dose rhenium-186-labeling of monoclonal antibodies for clinical application: pitfalls and solutions. AB - BACKGROUND: Rhenium-186 (186Re) has ideal properties for adjuvant radioimmunotherapy (RIT). However, the lack of suitable methods for high dose 186Re labeling of monoclonal antibodies (MAbs) has hampered the use of 186Re in clinical RIT. After development of a chemically identical multistep procedure for the production of 186Re-MAG3-MAb and 99mTc/99Tc-MAG3-MAb conjugates for use as a matched pair, the authors now report on further progress to make this labeling method broadly applicable for high dose 186Re labeling. METHODS: The number of metal-MAG3 groups that can be coupled to a MAb without alteration of the biodistribution was investigated by radioimmunoscintigraphy (RIS) in patients with head and neck squamous cell carcinoma (HNSCC). For labeling with 500 mCi [186Re]ReO4-, an efficient chemoprotection was introduced to suppress the damaging effects of radiation during conjugation and conjugate purification. Furthermore, the authors developed strategies that make the procedure easy and safe to perform at any medical center. RESULTS: MAbs showed a minor variation in biodistribution in HNSCC patients when the number of metal-chelate groups per MAb varied between < 1 and 4.3. High dose 186Re-MAb conjugates (150-250 mCi) with a Re-MAG3:MAb ratio of 3.4 were obtained with a radiochemical purity of > 95% and minimal aggregate formation (< or = 6%). Furthermore, a semiautomated labeling device and a convenient 100 mCi labeling kit in the form of a dried 186Re-MAG3 TFP ester were developed. RIT studies in HNSCC-bearing nude mice showed that high dose 186Re-labeled MAb U36 is more effective than iodine-131-labeled MAb U36. CONCLUSIONS: High dose 186Re-MAb conjugates were prepared that exhibit an optimal stability, immunoreactivity, and pharmacokinetic behavior. The availability of a kit procedure for coupling 186Re to MAbs might open the possibility of a broad application of 186Re in RIT. PMID- 9406685 TI - Experimental tumor targeting with radiolabeled ligands. AB - BACKGROUND: Approaches have been developed in animal models to increase the localization of radiolabeled ligands (monoclonal antibodies and peptides) in tumors, to reduce their uptake in normal tissues, and to thus improve the tumor/normal tissue uptake ratios so that higher and more frequent doses of radionuclide could be used for radioimmunotherapy. METHODS: These approaches to increase the localization of radiolabeled ligands in tumors involve the following three general strategies: (1) modifying ligands or radiolabeling techniques, (2) increasing blood and normal tissue clearance of radiolabeled ligands, and (3) modifying tumor delivery, tumor antigen, or receptor expression or increasing tumor vascular permeability or blood flow. RESULTS: The use of such animal models permits the assessment of a wide range of ligands, radiolabeling conditions, and the efficacy of administration methods before their initial use in clinical trials. The prospects for the use of radiolabeled ligands in cancer detection and therapy are promising because of their specificity for binding to receptors on tumor cells or tumor endothelial cells. CONCLUSIONS: Methods that increase the localization of radiolabeled ligands in solid tumors while reducing uptake in normal tissues will be required so that a sufficient radiation absorbed dose can be delivered for potentially curative treatment of radioresistant tumors in clinical radioimmunotherapy trials. PMID- 9406686 TI - Tumor targeting potential of the monoclonal antibody BC-1 against oncofetal fibronectin in nude mice bearing human tumor implants. AB - BACKGROUND: The immunoglobulin G1 (IgG1) monoclonal antibody (MoAb) BC-1 detects human oncofetal fibronectin, which has extremely restricted distribution in normal adult tissues and is highly expressed in fetal and tumor tissues. METHODS: We studied the biodistribution of 125I-labeled MoAb BC-1 in nude mice bearing subcutaneous human tumor implants of U87MG high-grade astrocytoma and SKMel28 melanoma. 125I-BC-1 was injected either intraperitoneally (i.p.) or intravenously (i.v.), and biodistribution was measured up to 144 hours after injection. In animals bearing SKMel28 implants, tumor targeting was also evaluated by in vivo imaging of the whole mouse by using a dedicated device based on transmitted light excitation after i.v. injection of MoAb BC-1 conjugated with the infrared fluorophore, CY7-bis(N-hydroxy-succinimido)-ester. RESULTS: 125I-BC-1 showed favorable uptake in the human tumor implants, reaching a maximum of 5.27 +/- 0.48% ID/g in the U87MG astrocytoma (72 hours after i.p. injection). The highest uptake in the SKMel28 melanoma implants was 3.49 +/- 0.25% ID/g (24 hours after i.v. injection). Microautoradiography of tumor specimens obtained after administration of 125I-BC-1 clearly showed radioactivity uptake within the two tumors replicating the same pattern of distribution as that of the oncofetal fibronectin shown by immunohistochemistry with MoAb BC-1. Nonspecific uptake of 125I-BC-1 in the bone marrow and skeletal muscle was much lower than in the tumors. In vivo imaging with the fluorophore-labeled MoAb clearly visualized the tumor implants 72-120 hours after i.v. injection. CONCLUSIONS: The experimental results obtained in this study demonstrate the favorable tumor targeting potential in vivo of the radiolabeled MoAb BC-1, a useful marker of neo angiogenesis induced by cancer. PMID- 9406687 TI - The potential of radiolabeled EGF-dextran conjugates in the treatment of urinary bladder carcinoma. AB - BACKGROUND: Muscle-invasive urothelial carcinoma of the urinary bladder has a poor prognosis in spite of available therapies. These tumors frequently overexpress the epidermal growth factor receptor (EGFr), a possible target for therapeutic conjugates. The aim of this study was to construct an EGF carbohydrate conjugate that would be potentially useful for both local (i.e., intravesical) and systemic radiotherapy. METHODS: EGF was coupled to periodate activated dextran by reductive amination. Receptor binding tests in vitro were conducted, using the human urothelial carcinoma cell line RT4 and the human malignant glioma cell line U-343-MG as a positive control. In the in vivo experiments, nude mice with subcutaneously grown xenografts of the RT4 tumor were injected intravenously with technetium-99m-labeled EGF conjugates. Samples of organs, blood, and tumors were collected after 24 hours. The radioactive uptake was calculated as a percentage of the dose per gram of tissue. RESULTS: The specific binding in the in vitro experiment was 90-95%. The binding was similar in both cell lines. There was a positive uptake in the tumors in the in vivo experiment, with tumor-to-blood ratios from 2:1 to 6:1. The uptake level in the kidneys was similar to that in the tumors (i.e., approximately 0.05% dose per gram of tissue). The other organs had a lower uptake compared with the tumor uptake. CONCLUSIONS: The results indicate that radiolabeled EGF-dextran has the potential to become a tool for local treatment of recurrent bladder carcinoma. With appropriate modifications, it should be possible to use this conjugate for systemic radiotherapy as well. PMID- 9406688 TI - The effect of antibody protein dose of anti-renal cell carcinoma monoclonal antibodies in nude mice with renal cell carcinoma xenografts. AB - BACKGROUND: Antibodies preferentially can direct radionuclides to solid tumors. However, antibody uptake in tumors is often highly heterogeneous. This heterogeneity may be overcome by increasing antibody protein dose. METHODS: The biodistribution of increasing protein doses of radioiodinated antirenal cell carcinoma (RCC) monoclonal antibodies (MoAbs) G250 and RC 38 was studied in mice with NU-12 or SK-RC-52 RCC xenografts. In addition, MoAb affinity constants and antigen densities (Scatchard analysis) and MoAb processing (internalization) were determined in vitro. RESULTS: The relative uptake of G250 in NU-12 tumors was very high at low protein doses (125% injected dose/g [%ID/g]), but decreased at higher doses, suggesting tumor saturation. Indeed, saturation of G250 antigen occurred at 3 microg protein. In this model, 9200 G250 determinants per NU-12 cell could be targeted, which is only 6.1% of the 150,000 G250 determinants per NU-12 cell as determined in vitro. The RC 38 uptake in NU-12 tumors remained constant up to the 10 microg dose level (40% ID/g) and decreased at higher doses. RC 38 antigens were saturated at 25 microg of RC 38. With RC 38, 15% of the available RC 38 antigens per NU-12 tumor cell were targeted. In contrast, G250 uptake in SK-RC-52 tumors was very low at low antibody dose (4% ID/g at 1 microg) and increased with increasing protein dose. These differences in G250 biodistribution might be related to differences in the processing of G250 by the tumor cells. CONCLUSIONS: Our studies show that some RCC tumors can be saturated with anti-RCC MoAbs at low (25 microg) to very low (3 microg) protein doses. At nonsaturated doses relatively high tumor uptake can be achieved. Surprisingly, in NU-12 tumors only 6.1% and 15% of the available antigenic sites were targeted at the saturating dose levels with G250 and RC 38, respectively. PMID- 9406689 TI - Radioimmunoscintigraphy with a novel monoclonal antiprostate antibody (E4): an experimental study in nude mice. AB - BACKGROUND: Prostate cancer is one of the leading causes of death among men, despite achievements in diagnosis and therapy. Radioimmunolocalization and radioimmunotherapy of malignant tumors have demonstrated increasing potential and may become useful tools in the management of prostate cancer. METHODS: Nude mice were inoculated subcutaneously with cells from the poorly differentiated human prostate cancer cell line DU-145. The intact monoclonal antibody (MoAb) E4 and an intact anticytokeratin-8 MoAb, TS1, used for comparison were labeled with 125I and injected intraperitoneally (i.p.) in the mice. Repetitive quantitative scintigraphic recordings were performed during 1 month. The mice were killed at Day 29 after injection of the radiolabeled MoAb. The tumors and the organs were dissected and weighed. The remaining activity was measured in a gamma well counter. One part of the tumor was immediately fixed in Bouin's solution for autoradiography and the other in formaldehyde for microscopy. RESULTS: The study demonstrated significant radioimmunolocalization of the MoAb E4 into the DU-145 prostate tumor tissue in the animal model, with an average radiation dose of 0.08 Gy/MBq in the tumor. TS1 localized preferentially in necrotic parts of the tumor, yielding a tumor dose of 0.02 Gy/MBq. CONCLUSIONS: The MoAb E4 is a promising radiotracer for prostate cancer and may be used in radioimmunotherapy. As in earlier studies, TS1 shows significant radioimmunolocalization into necrotic tumor tissue, which also exists in prostate cancer. PMID- 9406690 TI - Endogenous anti-tumor antibody responses in nude mice. AB - BACKGROUND: Nude mice with xenografted human tumors is the most exploited animal model used to elucidate the efficacy of experimental radioimmunolocalization and radioimmunotherapy. These animals accept transplants and are generally considered immunologically inert with regard to cell-mediated and humoral immune responses against the tumors. METHODS: Nude control mice and mice carrying human HeLa Hep-2 tumor xenografts were studied for appearance of endogenous antibodies following inoculation with tumor cells. The titers of these antibodies were investigated by isotype-specific enzyme-linked immunosorbent assay (ELISA) technologies, fluorescence-activated cell sorter analysis (FACS), BIAcore (Pharmacia Biosensor AB, Uppsala, Sweden) technology, and immunofluorescence. RESULTS: The HeLa Hep-2 cell line was found to be immunogenic in all investigated animals by means of ELISA, FACS, and BIAcore evaluations as well as by immunofluorescence against both tested antigens, placental alkaline phosphatase and cytokeratin 8. Predominantly immunoglobulin M antibodies were induced, but immunoglobulin G isotypes could also be identified. Sera from these tumor-bearing mice were used for immunohistochemistry of the tumor cells. The antibodies seemed to be of low affinity and may be displaced by high-affinity monoclonal antibodies used in radioimmunotargeting. CONCLUSIONS: Nude mice bearing tumor xenografts produce significant amounts of antibodies against these two human tumor-derived antigens. These endogenous antibodies may influence targeting of radiolabeled antibodies. They also have the potential to interfere with the pharmacokinetics of labeled or nonlabeled idiotypic antibodies during experimental immunolocalization. PMID- 9406691 TI - Improving tumor-to-normal-tissue ratios of antibodies by extracorporeal immunoadsorption based on the avidin-biotin concept: development of a new treatment strategy applied to monoclonal antibodies murine L6 and chimeric BR96. AB - BACKGROUND: Several strategies have been explored to accelerate monoclonal antibody (MAb) conjugate clearance without affecting intratumoral uptake. One of the most promising is extracorporeal immunoadsorption (ECIA), in which excess radiolabeled MAb circulating in blood is removed. METHODS: Extracorporeal immunoadsorption (ECIA) based on the avidin-biotin concept enables direct adsorption of radiolabeled and biotinylated MAb from plasma and consequently increases the tumor-to-normal-tissue uptake ratio by reducing background radioactivity in all radiosensitive organs. Because the concept is based on an antibody's being biotinylated prior to injection, the blood clearance efficiency is independent of the idiotype or isotype of the antibody employed. As a result, there is no need to develop new adsorption columns for each antibody system used. We have technically simplified the method recently by removing biotinylated MAb directly from blood without separation from plasma preceding the removal. The current study focused on both the development of ECIA and evaluating the effects of ECIA in terms of tumor targeting with two biotinylated radiolabeled MAbs that have different biokinetics, namely, murine MAb L6 and chimeric Mab BR96. RESULTS: The start time of ECIA should be determined for each MAb individually before radioimmunotherapy and be based on previous tumor biokinetics. BR96 with rapid tumor targeting seems more suitable than L6 for the ECIA procedure; it allows the procedure to start earlier and thereby further reduce whole body activity and exposure of critical organs to radiation. CONCLUSIONS: ECIA enables direct adsorption of radiolabeled and biotinylated monoclonal antibody from blood and consequently increases the tumor-to-normal-tissue uptake ratio. The method is even applicable to both the internalizing and already highly tumor-selective BR96 in a syngeneic tumor model. PMID- 9406692 TI - Tumor localization of a radiolabeled bombesin analogue in mice bearing human ovarian tumors induced to express the gastrin-releasing peptide receptor by an adenoviral vector. AB - BACKGROUND: The adenoviral vector, AdCMVGRPr, has been used to induce the expression of the murine gastrin-releasing peptide receptor (GRPr) both in vitro and in vivo. A bombesin analogue ([125I]-mIP-bombesin) has been shown to bind with high affinity to GRPr and to localize to intraperitoneal (i.p.) ovarian tumors 2 days after induction of GRPr in an athymic nude mouse model. The present study was conducted to determine the level of localization of [(125/131)I]-mIP bombesin in the tumors at 2, 4, and 7 days after AdCMVGRPr administration and to determine the feasibility of giving multiple doses of [131I]-mIP-bombesin for therapy. METHODS: Human ovarian cancer cells (SKOV3.ip1) were infected in vitro with AdCMVGRPr and were assayed for receptor expression at 2, 4, and 7 days after infection by using a radiolabeled bombesin-binding assay. Biodistribution studies utilized athymic nude mice inoculated i.p. with SKOV3.ip1 cells. The tumors were induced to express GRPr with an i.p. injection of AdCMVGRPr followed by administration of [125I]-mIP-bombesin 2 days later (AdCMVLacZ or saline was used for negative controls). In addition, the tumor localization of [125I]-mIP bombesin was determined 4 and 7 days after AdCMVGRPr administration. The tumor localization of [131I]-mIP-bombesin was compared with [125I]-mIP-bombesin in this in vivo model. RESULTS: SKOV3.ip1 cells infected with AdCMVGRPr resulted in 80.3 +/- 5.9% binding of [125I]-Tyr4-bombesin at 2 days after infection, which decreased to 46.8 +/- 0.4% at 4 days and to 17.7 +/- 0.1% at 7 days. The biodistribution study showed that the tumor localization (14.9 +/- 8.2% injected dose/gram; ID/g) of [125I]-mIP-bombesin 2 days after administration of AdCMVGRPr was significantly greater than its localization in other organs (P < 0.003) and was significantly greater than in AcCMVLacZ- and saline-treated mice (P < 0.003). Injections of [125I]-mIP-bombesin at 4 and 7 days after a single AdCMVGRPr administration showed tumor localization of 4.5 +/- 3.0% ID/g at Day 4 and 3.9 +/ 3.5% ID/g at Day 7. The decreased localization at longer times after AdCMVGRPr infection correlated with in vitro results. The tumor uptake of [125I]-mIP bombesin was comparable to the uptake of [131I]-mIP-bombesin (21.2 +/- 8.3% ID/g versus 15.4 +/- 5.6% ID/g, respectively), as was the normal tissue biodistribution. CONCLUSIONS: The expression of GRPr in human ovarian cancer cells can be accomplished both in vitro and in vivo by using AdCMVGRPr, with the in vivo tumor localization of [125I]-mIP-bombesin being significantly greater than in control animals. The tumor localization of [125I]-mIP-bombesin and [131I] mIP-bombesin at 2 days after AdCMVGRPr was comparable in a mouse model of human ovarian carcinoma. Injections of [125I]-mIP-bombesin at Days 4 and 7 after AdCMVGRPr infection resulted in tumor localization of [125I]-mIP-bombesin but at a level lower than 2 days. Thus, the total amount of radioactivity delivered to the tumor should be increased by multiple injections of [131I]-mIP-bombesin, which would be required for a therapeutic effect. PMID- 9406693 TI - Processing of antibodies bound to B-cell lymphomas and lymphoblastoid cell lines. AB - BACKGROUND: Previous experiments demonstrated that some human B-cell lymphoma cell lines were unusual in that antibodies bound to the cell surface dissociated at high levels. This did not occur with non-B-cell hematologic tumors or with carcinomas. In this study, additional B-cell lymphoma and lymphoblastoid (Epstein Barr virus-transformed) cell lines were tested. METHODS: The antibodies selected for most experiments, MA103 and anti-CD45, react with relatively high avidity to the cell surface. Antibodies to CD19, CD20, and CD22 also were tested on certain cell lines. The antibodies were labeled with 125I. After binding to the surface of viable cells, unbound antibody was washed away, and the fate of the bound antibody was investigated for 2-3 days. RESULTS: Of the eight B-cell lymphomas tested, three had high levels of dissociation, two had low levels of dissociation, and three had intermediate levels of dissociation. The six lymphoblastoid cell lines had only slightly elevated levels of dissociation, relative to non-B cell lines. Sublines of Raji and Ramos cells were identified that varied greatly in the level of antibody dissociation. The level of dissociation from lymphomas was correlated with the tendency of the cell lines to cluster, with single cells displaying less dissociation than clustered cells. However, some exceptions to this correlation were noted. Cell lines such as Ramos, which showed little dissociation of anti-CD20, displayed relatively rapid catabolism of this antibody. CONCLUSIONS: The level of antibody dissociation as well as the rate of antibody catabolism will affect the results of radioimmunotherapy strongly because these factors affect the time interval for which the cells are in contact with the radioisotope. Different B-cell lines display markedly different levels of dissociation. There is some evidence suggesting that antibody dissociation is high with fresh human tumor cells, but further investigation of this point is required. PMID- 9406694 TI - Perspectives on oncologic imaging with radiolabeled antibodies. AB - BACKGROUND: The contributions of nuclear imaging tests to the management of cancer are expanding, especially in terms of detection (staging and recurrence), diagnosis, and qualification of patients for certain forms of therapy, and particularly with regard to the tests' ability to identify tumors on a functional basis. METHODS: This article is a selective review of the advances and clinical management applications of cancer imaging with radiolabeled antibodies (radioimmunodetection or immunoscintigraphy), with the objective of demonstrating how this new imaging technology can contribute to oncologic practice. The contribution of another functional imaging modality, positron emission tomography (PET), also is discussed. RESULTS: Radioimmunodetection and PET imaging have shown particular management contributions in patients with colorectal, breast, or lung carcinoma, as well as in patients with non-Hodgkin's lymphoma (NHL). Two antibody imaging agents, OncoScint and CEA-Scan, currently are available for the detection of colorectal carcinoma, and have shown particular application for disease staging and disclosure of occult lesions. Each of these agents has different characteristics and potential applications. Recent results with CEA Scan in patients with mammary carcinoma suggest that it may be a useful complementary tool to screening mammography by adding very high specificity. In patients with lung carcinoma, technetium-99m-labeled NR-LU-10 antibody has been shown to be useful in preoperative staging. LymphoScan is under investigation as a staging agent and for revealing residual disease in patients with NHL; it also provides, with a different radioisotope attached and in a humanized antibody form, a therapeutic option for patients with NHL. Similar results can be shown for PET, but this method may not provide the specificity of an antibody-based functional imaging agent, although PET may have a higher lesion sensitivity. CONCLUSIONS: Both radioimmunodetection and PET provide new exciting opportunities for earlier and more sensitive functional imaging tests for cancer. In contrast to PET, antibody agents also can serve to qualify a patient as a candidate for an antibody-based therapy. PMID- 9406695 TI - Anti-carcinoembryonic antigen antibodies versus somatostatin analogs in the detection of metastatic medullary thyroid carcinoma: are carcinoembryonic antigen and somatostatin receptor expression prognostic factors? AB - BACKGROUND: Surgery is currently the only potentially curative approach in the treatment of medullary thyroid carcinoma (MTC). In many instances however, postsurgically elevated or rising plasma calcitonin and/or carcinoembryonic antigen (CEA) levels indicate persistent metastatic disease, although conventional diagnostic procedures (computed tomography (CT), magnetic resonance imaging (MRI), and invasive venous catheterization) fail to localize the responsible lesions. Recently, anti-CEA antibodies and somatostatin analogs have shown promising results in the staging of MTC. The aim of this study was to compare the sensitivity of both methodologies, especially for the detection of occult MTC, and to assess whether there may be correlations between the scintigraphic behavior and the patients' prognosis. METHODS: A total of 26 patients with medullary thyroid carcinoma were examined at our institution between 1977 and 1996. Ten of them had known disease, 14 had occult metastatic MTC, and 2 were free of disease at the time of presentation. Fourteen patients were investigated with anti-CEA monoclonal antibodies (MAbs) (receiving a total of 35 injections: clones BW431/26, BW431/31, IMACIS, or F023C5, labeled with 99mTc, (111)In or (131)I), and 8 patients were studied with (111)In-labeled octreotide. Two patients received potentially therapeutic doses of (131)I-labeled anti-CEA antibodies. All patients underwent conventional radiologic evaluation (ultrasonography, CT, and MRI) and/or biopsy within 4 weeks. Additional imaging was performed with 99mTc-(V)-DMSA, (131)I-metaiodobenzylguanidine, 201thallium chloride, 99mTc-methylene diphosphate, and/or 18F-fluorodeoxyglucose-positron emission tomography. Clinical follow-up was obtained. RESULTS: All patients with established disease had elevated plasma CEA (range, 6.8-345 ng/mL; calcitonin levels between 92 and 11,497 pg/mL), whereas in 9 of 14 occult cases, levels were < or = 5 ng/mL (range, 0.6-829 ng/mL; calcitonin, 72-2920 pg/mL). In patients with known disease, the overall lesion-based sensitivity was 86% for the anti-CEA MAbs, whereas octreotide was unable to target any tumor in patients with rapidly progressing disease or distant metastases (overall sensitivity, 47%). In all patients with occult MTC, anti-CEA MAbs and octreotide were able to localize at least one lesion (patient-based sensitivity, virtually 100%). In patients with postsurgically persistent hypercalcitoninemia, cervical lymph nodes were identified as the most frequent site of metastases, whereas in patients with occult and slowly progressing disease several years after primary surgery, anti CEA MAbs and octreotide showed bilateral involvement of mediastinal lymph nodes; however, tumor to nontumor ratios were usually higher with octreotide in these cases. With anti-CEA Mabs, the highest tumor to nontumor ratios were observed in clinically aggressive, rapidly progressing disease. The sensitivity of all other diagnostic modalities was, at < or = 50%, significantly lower. Indication for antitumor effects was observed in a patient receiving 65 mCi of (111)I-labeled F(ab')2 fragments of the clone F023C5. CONCLUSIONS: For the detection of occult MTC, anti-CEA MAbs and octreotide seem to have a sensitivity that is superior to conventional diagnostic modalities, especially also when used in combination. Better detectability with anti-CEA antibodies (which may result in higher CEA expression) seems to be associated with more aggressively growing forms of MTC, whereas somatostatin receptor expression at normal CEA plasma levels and weaker MAb targeting may be associated with a more benign clinical course. This is in accordance with the study of Busnardo et al. (Cancer 1984; 53:278-85), who showed higher CEA serum levels to be associated with a worse prognosis, as well as with the in vitro findings of Reubi et al. (Lab Invest 1991;64:567-73), who demonstrated lower somatostatin receptor expression in less differentiated MTC. Fu PMID- 9406696 TI - Single chain antigen binding protein (sFv CC49): first human studies in colorectal carcinoma metastatic to liver. AB - BACKGROUND: An sFv fragment of the anti-TAG-72 monoclonal antibody CC49 has been developed and has shown promise in improved targeting to colorectal carcinoma in animal studies. In this study the authors report their initial experience in human patients after intravenous injection. METHODS: Five patients with colorectal carcinoma metastatic to the liver were studied prior to surgery. High performance liquid chromatography showed a low level of aggregation (< 10% complex formation), before and after radiolabeling with iodogen. Prior to radiolabeling, 123I was brought to the dry form, phosphate buffer added and titrated to a pH of 7, with diluted hydrochloric acid. 123I was injected in doses of 26, 12, 27, 25 and 1 millicurie, respectively, and labeled to a 5-mg fragment. Single photon emission computed tomography and whole body imaging were performed at 4-6 hours, and 24 hours, respectively, after injection. RESULTS: The agent was rapidly cleared from the blood with biphasic clearance T-1/2 of 30 minutes and 10.5 hours, respectively. Distribution from whole body imaging confirmed rapid equilibration with extracellular fluid, and clearance T-1/2 from the body was comparable to the slower component of blood clearance. The spleen was visualized in all patients, and the testes were imaged in 67% of male patients. Renal excretion was noted with early uptake and clearance from the renal parenchyma except in one patient in whom renal parenchyma retention was intense. Although image quality was suboptimal, tumor was visualized in all five patients in both primary and metastatic lesions. At surgery, (16-24 hours postinjection), the tumor retained significant concentrations of the radiotracer, with metastatic tumor/normal liver ratios of approximately 1:5-3:1. No patient had any associated symptom or change in biochemical and hematopoietic status. CONCLUSIONS: This study showed that sFv is safe, tissue equilibration and clearance is rapid, and early, same-day imaging of the primary and metastatic tumors is feasible in patients colorectal carcinoma. Further studies are warranted to define a more optimal mass of sFv CC49 dose for tumor targeting. PMID- 9406697 TI - Tc-99m LL-2 Fab' monoclonal antibody imaging in acquired immune deficiency syndrome-related lymphoma. AB - BACKGROUND: Both systemic and primary central nervous system (CNS) non-Hodgkin's lymphomas (NHL) occur in people with acquired immune deficiency syndrome (AIDS). The radiographic manifestations may be similar to other neoplasms and opportunistic infections that are also found frequently in AIDS. Furthermore, these diseases may coexist with NHL in the AIDS patient. METHODS: To evaluate the use of Tc-99m Lymphoscan (the Fab' fragment of the anti-CD-22 antibody LL-2; Immunomedics, Inc., Morris Plains, NJ) in patients with suspected AIDS lymphoma, we studied 7 patients with 35 sites of suspected disease. Six had CNS lesions suspicious for parenchymal brain lymphoma. Each patient underwent planar and single photon emission computed tomography imaging at 3-5 and 18-24 hours after administration of Lymphoscan. Scintigraphic results were compared with results of conventional diagnostic modalities. RESULTS: Overall, the sensitivity of Lymphoscan was 92% and the specificity was 86%. In brain lesions, there was 100% sensitivity and 100% specificity. Lymphoscan also had 100% sensitivity for sites of lymphomatous lymphadenopathy and for liver involvement. Although less specific in extracranial sites, Lymphoscan was correctly negative in sites of coexisting adenocarcinoma and pneumonia. Two patients had both parenchymal CNS and systemic lymphoma proven by biopsy. CONCLUSIONS: Lymphoscan appears to be a sensitive and specific method for diagnosing CNS lymphoma in AIDS patients. Although slightly less specific in extracranial sites, it may be helpful in differentiating lymphoma from other etiologies in these patients at risk for multiple neoplasms and opportunistic infections. PMID- 9406698 TI - Comparison of Technetium-99m sestamibi and indium-111 octreotide imaging in a patient with Ewing's sarcoma before and after stem cell transplantation. AB - BACKGROUND: We report the use of two novel nuclide agents, Technetium-99m (99Tc)sestamibi (MIBI) and indium-111 (In-111) octreotide, in comparison with conventional computed tomography (CT) imaging in a patient with metastatic Ewing's sarcoma (ES) before and after high dose chemotherapy with autologous peripheral stem cell transplantation (PSCT). MIBI is taken up actively by metabolically active tumor cells. Octreotide, a somatostatin analog, binds specifically to somatostatin receptors. METHODS: The patient was a 20-year-old male with recurrent metastatic ES to the lung. Before and sequentially after high dose chemotherapy and PSCT, the patient was imaged with MIBI. Whole body planar and single photon emission computed tomography (SPECT) images were obtained after the injection of 30 mCi of 99Tc MIBI. Prior to PSCT the patient was imaged with 6 mCi In-111 pentreotide. RESULTS: Conventional CT scans also were performed. Initial CT revealed pulmonary metastasis in the right lower lobe along with multiple left pleural-based lesions. These lesions were visualized clearly with MIBI. Octreotide detected only the left lung involvement. Sequential MIBI scans after PSCT correlated with tumor reduction in the right lung field and tumor progression in the left lung as well as the development of new pulmonary metastasis. These findings were confirmed on CT. CONCLUSIONS: MIBI imaging was highly concordant with CT scanning in the detection of metastatic ES. MIBI scanning holds promise for the direct detection of a variety of human malignancies, and may prove useful as a rapid whole body imaging modality. PMID- 9406700 TI - Intraoperative gamma detection reveals abdominal endocrine tumors more efficiently than somatostatin receptor scintigraphy. AB - BACKGROUND: [(111)In]-diethylenetriamine pentaacetic acid (DTPA)-D-[Phe1] octreotide (OctreoScan, Mallinckrodt Medical, Petten, The Netherlands), combined with single photon emission computed tomography (SPECT), has limited possibilities to detect small abdominal endocrine tumors. This study explores the efficacy of OctreoScan and a newly developed handheld gamma detector probe (H probe2) for the localization of neuroendocrine abdominal tumors. METHODS: Twenty one patients with endocrine pancreatic or midgut carcinoid tumors were given 100 200 megabecquerels (MBq) [(111)In]-DTPA-D-[Phe1]-octreotide and underwent preoperative SPECT examination, after which time surgery was undertaken within 24 48 hours. Intraoperatively the radioactivity of tumors and normal tissues was measured with the H-probe2 connected to a portable personal computer. Resected intestinal specimens of ten patients with carcinoid tumors were examined by gamma camera ex vivo. Biopsies of all tumors subsequently were investigated in a wellcounter and by routine histopathology. RESULTS: Disregarding liver metastases, a total of 34 of 60 abdominal tumors were detected by SPECT; however, SPECT failed to visualize any tumor < 9 mm. The shielding of the manual probe allowed intraoperative examination despite the substantial radiation energy of (111)In. The H-probe2 detected 91% of the tumors investigated, and these included all those > 5 mm. In the wellcounter the H-probe2-detected tumors exhibited 4.7 157.7 times higher radionuclide uptake than the surrounding normal tissues. The gamma camera analysis detected 68% of tumors in the resected specimens. CONCLUSIONS: The intraoperative H-probe2 examination provided substantial improvement in the detection of small endocrine lesions accumulating [(111)In] DTPA-D-[Phe1]-octreotide and all lesions with a size > 5 mm in greatest dimension could be identified. PMID- 9406699 TI - A pilot pharmacokinetic and immunoscintigraphic study with the technetium-99m labeled monoclonal antibody BC-1 directed against oncofetal fibronectin in patients with brain tumors. AB - BACKGROUND: Preliminary experiments in an animal model have shown the favorable tumor targeting potential in vivo of radiolabeled BC-1, an immunoglobulin (Ig)G1 monoclonal antibody (MoAb) that recognizes the human fibronectin isoform (B+) containing the ED-B oncofetal domain. This antigen has extremely restricted distribution in normal adult tissues. Instead, it is highly expressed in fetal and tumor tissues, especially in high grade astrocytomas and malignant gliomas of the brain, in which the process of neoangiogenesis linked to tumor growth is particularly important. METHODS: This study was carried out with five patients who had malignant brain tumors (four gliomas and one malignant angioblastic meningioma). The BC-1 MoAb was labeled with technetium-99m (99mTc) by MDP transchelation. Planar and single photon emission computed tomography (SPECT) imaging was acquired at 4-6 and 20 hours after intravenous injection of about 450 MBq/0.2 mg 99mTc-BC-1 and was compared with the nonspecific indicator of blood brain barrier disruption, 99mTc-diethylenetriamine pentaacetic acid (DTPA). Plasma pharmacokinetic analysis was based on serial blood sampling. All patients underwent potentially curative surgery at the end of the study. RESULTS: The plasma clearance curves were biexponential, with average T(1/2) values of 2-4 hours and 28-33 hours, respectively. 99mTc-BC-1 showed very low nonspecific uptake in the bone marrow, liver, and spleen. Planar and SPECT imaging with 99mTc BC-1 visualized brain tumors in all patients, with a pattern of intratumor distribution that specifically identified areas of peripheral tumor growth more accurately than the nonspecific indicator, 99mTc-DTPA. Tumor uptake of 99mTc-BC-1 was correlated with the expression of the specific oncofetal fibronectin, as shown by immunohistochemistry on surgical samples. CONCLUSIONS: These results indicate the diagnostic potential of MoAb 99mTc-BC-1 for immunoscintigraphy in cancer patients, at least when neoangiogenesis induced by cancer is particularly important. PMID- 9406701 TI - A gamma detector probe with ex vivo detection of carcinoid tumors superior to intraoperative palpation. AB - BACKGROUND: Despite the generally successful scintigraphic detection of endocrine tumors with [(111)In]-DTPA-D-[Phe1]-octreotide (OctreoScan, Mallinckrodt Medical, Petten, The Netherlands), its intraoperative application awaits the development of suitable gamma detectors. This study describes a novel probe (H-probe2) and its ex vivo efficacy for the detection of midgut carcinoid tumors. METHODS: The probe measures 180 mm x 24 mm, and contains a bismuth germinate crystal connected to a photomultiplicator tube, a lead shield, and a tantalum collimator with an angled, 3-mm wide opening. It was characterized in a test bench utilizing solublilized (111)In and 99mTc, and 8 fresh operative specimens containing 26 histologically verified midgut carcinoid tumors (2-40 mm) from patients exposed to OctreoScan. Measurements were made at 2.5-mm intervals over the entire specimens and presented three dimensionally. RESULTS: The test bench analysis supported efficient shielding, and a total collimation of 27 degrees for (111)In and 20 degrees for 99mTc. In addition to 18 palpable tumors, 4 of 6 surgically occult tumors (2-3 mm in dimension) could be discovered with H-probe2. Small tumors in close apposition to a large tumor could not be separated. Wellcounter examination showed that tumors detected with the H-probe2 exhibited 1.7-84.1 times higher radionuclide uptake than the surrounding normal tissue. CONCLUSION: Ex vivo analysis of H-probe2 supported detection of tumors inaccessible to surgical palpation, but its clinical efficiency awaits intraoperative evaluation. PMID- 9406702 TI - Dosimetry overview: keeping score on the scorekeepers. AB - BACKGROUND: As a direct result of the use of the absorbed dose unit the 'Gray' as the gold standard for predicting response in external beam radiotherapy, the physicist role has been essential to clinical practice for many decades. However, although the dosimetry for internal emitters has proven useful in managing health physics concerns and diagnostic nuclear medicine, the relative success of correlating absorbed dose with response from radionuclide therapy has been limited. METHODS: This overview presents the relative success and/or failure of model-based dosimetry for radionuclide therapy in comparison to results quoted for external beam therapy dosimetry. RESULTS: Using the standard MIRD formalism for macroscopic dosimetry, the marked non-uniform distribution of radionuclide in both tumor and normal tissue has resulted in limited correlation between computed absorbed dose and biological response in clinical trials. Several efforts are underway aimed at improving this dose-response correlation which include individualized patient specific dosimetry and selected biological parameters. CONCLUSIONS: The physicist role in helping the clinician determining which patients will succeed on given radionuclide therapy has been improved with simplified methods such as the assessment of tracer whole body absorbed dose on a per patient basis. The dose-response correlations are now in the moderate range of significance when individualized patient dosimetry is included. These correlations are expected to improve as unified treatment planning programs are instituted and standard methods of clinically based dosimetry are widely accepted and practiced universally. PMID- 9406703 TI - Segmentation of lung lesion volume by adaptive positron emission tomography image thresholding. AB - BACKGROUND: It is common protocol in radionuclide therapies to administer a tracer dose of a radiopharmaceutical, determine its lesion uptake and biodistribution by gamma imaging, and then use this information to determine the most effective therapeutic dose. This treatment planning approach can be used to quantitate accurately the activity and volume of lesions and organs with positron emission tomography (PET). In this article, the authors focus on the specification of appropriate volumes of interest (VoI) using PET in association with computed tomography (CT). METHODS: The authors have developed an automatic image segmentation schema to determine the VoI of metastases to the lung from PET images, under conditions of variable background activity. An elliptical Jaszczak phantom containing a set of spheres with volumes ranging from 0.4 to 5.5 mL was filled with F-18 activity (2-3 microCi/mL) corresponding to activities clinically observed in lung lesions. Images were acquired with a cold background and then with variable source-to-background (S/B) ratios of: 7.4, 5.5, 3.1, and 2.8. Lesion VoI analysis was performed on 10 patients with 17 primary or metastatic lung lesions, applying the optimum threshold values derived from the phantom experiments. Initial volume estimates for lung lesions were determined from CT images. Approximate S/B ratios were obtained for the corresponding lesions on F 18-fluoro-2-deoxy-D-glucose (18FDG)-PET images. From the CT estimate of the lesion size and the PET estimate of the S/B ratio, the appropriate optimum threshold could be chosen. The threshold was applied to the PET images to obtain lesion activity and a final estimate of the lesion volume. RESULTS: Phantom data analysis showed that image segmentation converged to a fixed threshold value (from 36% to 44%) for sphere volumes larger than 4 mL, with the exact value depending on the S/B ratios. For patients, the use of optimum threshold schema demonstrated a good correlation (r = 0.999) between the initial volume from CT and the final volume derived from the 18FDG-PET scan (P < 0.02). The mean difference for those volumes was 8.4%. CONCLUSIONS: The adaptive thresholding method applied to PET scans enables the definition of tumor VoI, which hopefully leads to accurate tumor dosimetry. This method can also be applied to small lesions (<4 mL). It should enable physicians to track objectively changes in disease status that could otherwise be obscured by the uncertainties in the region-of-interest drawing, even when the scans are delineated by the same physician. PMID- 9406704 TI - Dosimetry of fractionated administration of 125I-labeled antibody at experimental radioimmunotargeting. AB - BACKGROUND: Radiotherapy of solid tumors is preferably performed in fractionated doses. Conversely, radioimmunotherapy with nuclide-carrying antibodies delivers a continuously decreasing low dose rate during a longer time period after a single injection. In the current study, the same total amount of 125I-labeled anticytokeratin monoclonal antibody (MoAb) was administrated in one, three, or ten injections and the dosimetry was evaluated. METHODS: Three groups of nude mice (10 mice each) with HeLa Hep 2 xenografts were injected with 1 x 100 microg/22.2 megabecquerel (MBq), 3 x 33 microg/7.4 MBq, and 10 x 10 microg/2.22 MBq 125I-labeled TS1 MoAb, respectively. The mice were examined scintigraphically over a 54-day period (total number of radio immunoscintigraphies (RISs) = approximately 700) and doses to tumor and normal tissues were estimated according to the medical internal radiation dose formalism. RESULTS: A single bolus injection caused higher tumor uptake, tumor dose, and tumor to nontumor dose ratio than administration of the same total dose of antibody and radioactivity in three or ten separate injections. The single bolus injection caused a tenfold higher tumor uptake (% injected dose, or ID) compared with the group receiving ten injections. This caused a tumor dose of 17 gray to the group receiving a single bolus injection. CONCLUSIONS: In this antigen target system, a single injection of a large amount of antibody was found to be more efficient than the same antibody dose subdivided into three or ten fractions. It was concluded that not only the radioactivity but also the amount of antibody per fraction should be considered when determining optimal fractionated radioimmunotherapy. PMID- 9406705 TI - Radiobiologic studies of radioimmunotherapy and external beam radiotherapy in vitro and in vivo in human renal cell carcinoma xenografts. AB - BACKGROUND: Previous studies suggest that the radiobiologic characteristics of in vitro survival curves are important determinants of the response of tumors to both conventional radiotherapy and radioimmunotherapy (RIT). The purpose of this study was to elucidate the relationship between in vitro radiation survival curve parameters and the relative sensitivity of tumor to RIT, exponentially decreasing low dose rate (ED LDR) irradiation and conventional high dose rate (HDR) fractionated external beam radiotherapy. METHODS: Two human renal cell carcinoma cell lines, Caki-1 and A498, were used in vitro and nude mouse xenograft studies. HDR external beam gamma irradiation (dose rate, 430 centigray [cGy]/minute) and ED LDR irradiation (initial dose rate, 22-25 cGy/hour) were performed with a cesium-137 (137Cs) gamma irradiator. RIT was carried out with yttrium-90 (90Y labeled monoclonal antibody NR-LU-10, and the absorbed radiation doses were calculated by medical internal radiation dose methodology. A clonogenic assay was used to generate radiation survival curves, and a computer FIT program was used to calculate the radiobiologic parameters. The antitumor efficacy of the different treatments was compared in vivo using a tumor regrowth delay assay in these two tumor xenograft models. RESULTS: The radiation survival curves showed that the Caki-1 cell line was more sensitive to both HDR and ED LDR irradiation than A498 in vitro. The Caki-1 cell line, compared with A498, had a larger alpha (0.39 vs. 0.15 Gy following HDR and 0.32 vs. 0.21 Gy following ED LDR) and alpha to-beta ratio (6.92 vs. 2.60 Gy for HDR and 40.0 vs. 19.2 Gy for ED LDR), a smaller n number (5.13 vs. 23 for HDR and 1.16 vs. 3.53 for ED LDR), a lower quasi-threshold dose (Dq) (1.60 vs. 3.15 Gy for HDR and 0.35 vs. 1.76 Gy for ED LDR), and a lower surviving fraction at 2 Gy (SF2) (0.37 vs. 0.60 for HDR and 0.51 vs. 0.61 for ED LDR), suggesting that Caki-1, compared with A498, had a steep initial slope and a small shoulder. The final slope represented by the beta value and D0 dose (the dose (Gy) required to reduce the fraction of surviving cells of 37% of its previous value in the exponential region of the survival curves) did not vary significantly between these two cell lines at either HDR or ED LDR irradiation. Tumor volume doubling times were 4.0 +/- 1.5 days for Caki-1 and 4.2 +/- 1.8 days for A498 tumor xenografts. One hundred microCi/50 microg of 90Y-labeled, isotype-matched irrelevant monoclonal antibody CCOO16-3 produced a tumor growth delay time (TGD) of 2.1 days in Caki-1 tumors but had no effect on A498 tumors (P < 0.05). RIT with 100 microCi of 90Y-NR-LU-10 resulted in a TGD of 4.8 days for Caki-1 tumors, whereas 100 microCi and 150 microCi of 90Y-NR-LU-10 produced a TGD of 1.9 and 2.7 days for A498 tumors, respectively. Estimated absorbed doses were 21.9 Gy in Caki-1 tumors treated with 100 microCi of 90Y-NR LU-10 and 14.5 Gy and 21.8 Gy in A498 tumors treated with 100 microCi and 150 microCi of 90Y-NR-LU-10, respectively. The weighted normal tissue absorbed doses were 7.4 Gy for Caki-1 tumor-bearing mice and 9.0 Gy for A498 tumor-bearing mice (P > 0.05). To compare the responses of Caki-1 and A498 xenografts to RIT with external beam ED LDR and HDR irradiation, tumor-bearing mice were treated with equivalent doses (20-22 Gy) of 1) RIT with 90Y-NR-LU-10 (100 microCi for Caki-1 and 150 microCi for A498), 2) continuous ED LDR 137Cs irradiation with a initial dose rate of 22 cGy/hour, or 3) HDR X-irradiation (2 Gy x 10 fractions in 2 weeks). The TGDs produced by RIT, ED LDR, and HDR were 5.3, 9.7, and 8.3 days for Caki-1 and 2.7, 5.1, and 5.8 days for A498. The relative efficacy of RIT in these xenograft models correlated well with the radiobiologic parameters (i.e., the size of the initial slope and shoulder) of in vitro survival curves following HDR and ED LDR irradiation in these cell lines. (ABSTRACT TRUNCATED) PMID- 9406706 TI - p53-independent response of a human breast carcinoma xenograft to radioimmunotherapy. AB - BACKGROUND: Radiation-induced DNA damage resulting in p53 protein attachment and downstream gene activation has been considered a major mechanism for tumor response to low dose rate radiation therapy. In this study, the mechanism of tumor response, and p53 gene status as well as levels of expression of p53 pathway genes were investigated in a human breast tumor (HBT 3477) before and after yttrium-90-DOTA-peptide-ChL6 (Y-90-ChL6) treatment of these xenografts. METHODS: Mice with HBT 3477 xenografts were treated with 260 microCi Y-90-ChL6 and sacrificed 3, 24 and 48 hours after injection. Reverse transcriptase polymerase chain reaction and/or Western blotting were used to measure the tumor levels of p53, p21(CDKN1/WAF1) (p21), GADD45, and bcl-2. Single strand conformation polymorphism and direct sequencing were used to determine the mutational status of p53. Evidence of apoptosis was determined by cleavage of poly(ADP-ribose) polymerase (PARP). RESULTS: Tumors regressed 4-7 days after treatment with 260 microCi Y-90-ChL6, resulting in a 79% tumor response. The p53 gene mutation found at codon 342 in HBT 3477 resulted in truncation of the p53 protein, and correlated with undetectable basal p21 protein levels. GADD45 and p53 mRNA decreased after therapy. bcl-2 mRNA was abundant, but decreased. Retinoblastoma phosphorylation showed no changes. Cleavage of PARP was detected at 3 hours and levels were increased greatly at 6 hours after therapy. CONCLUSIONS. Response in the Y-90-ChL6 treated HBT 3477 xenograft tumors was independent of p53 and occurred by apoptosis. The down-regulation of bcl-2 may be the key in this apoptotic response to low dose rate radioimmunotherapy. PMID- 9406707 TI - Preliminary results from intensity-based CT-SPECT fusion in I-131 anti-B1 monoclonal-antibody therapy of lymphoma. AB - BACKGROUND: In treatment of non-Hodgkin's lymphoma patients with predose-plus-I 131-labeled anti-B1 (anti-CD20) monoclonal antibody, an intratherapy single photon emission computed tomography (SPECT) image is an important part of research estimates of tumor dosimetry. For that imaging, a computed tomography (CT)-SPECT fusion is used both to obtain an attenuation map for the space alternating generalized expectation maximization reconstruction and to provide CT based volumes of interest (VoI) to determine activity in tumors and organs. Fusion based on external, skin-surface markers has been used but may not correctly superimpose internal structures. METHODS: A new algorithm, developed and implemented in the Department of Radiology, University of Michigan, and based on the mutual information of grayscale values, was investigated. Results from four anti-B1 therapy patients are presented. RESULTS: In one patient, the new intensity-based fusion provided total reconstructed counts for kidneys that were higher than those produced by marker-based fusion; therefore, the VoI was probably located more accurately. In a second patient, after an acquisition that did not include any skin markers, the new algorithm produced counts/pixel that were similar for four of five tumors consistent with what is expected from an ideal therapy combined with accurate count density estimates. The fifth tumor was quite small and will have its final activity estimate moved toward consistency with the others after a recovery coefficient multiplication. For four tumors in two patients, direct comparison of the two algorithms yielded count totals that were different by no more than 7.2%. CONCLUSIONS: The use of CT-SPECT fusion and subsequent transfer of tumor VoI originally drawn in high-resolution CT space offers potential advantages for quantifying tumor uptake of radioactivity. A new, mutual-information-based fusion algorithm is usable without skin markers. Results indicate that the new fusion algorithm gives equal tumor count values within 7.2% compared with fusion based on external markers. It increases estimates of kidney activity by an average of 6.4%. PMID- 9406708 TI - The effect of three dimensional activity distribution on the dose planning of radioimmunotherapy for patients with advanced intraperitoneal pseudomyxoma. AB - BACKGROUND: Six patients with histologically proven peritoneal carcinomatous pseudomyxomas were treated with radioimmunotherapy. METHODS: All the patients received a tracer dose of iodine-131 (131I) labeled B72.3 anti-TAG-72 monoclonal antibody (MoAb) to test the in vivo affinity. After informed consent was obtained the therapeutic dose (>3.7 gigabecquerels [GBq], 100 mCi) of the 131I labeled B72.3 anti-TAG-72 MoAb was infused within 60 minutes intraperitoneally using 2 catheters on both sides of the abdomen. The patients were imaged with single photon emission computed tomography (SPECT) at 3, 10, and 24 days after the therapeutic infusion. Treatment-planning software has been developed in which functional information obtained from SPECT is integrated with anatomic information obtained from computed tomography (CT). The activity distribution from SPECT images is converted to absorbed dose distributions using a point source kernel convolution dose calculation. The absorbed dose calculation requires a radionuclide specific dose kernel. The activity map is divided into equally sized source voxels from which the distribution is calculated for the target voxels that cover the patient volume. The resulting three dimensional (3D) absorbed dose distribution is viewed as isodose contours superimposed on the CT images or as 3D isodose surfaces. RESULTS: The measured activity distribution shows that the cumulated activity and biologic half-life vary in the patient's body. The developed planning system provides a method for calculating patient specific absorbed dose distributions. CONCLUSIONS: The variation of biologic clearance indicates that a 3D dose calculation method incorporating measured activity distributions is needed to quantify absorbed dose distribution. PMID- 9406709 TI - Impact of splenomegaly on therapeutic response and I-131-LYM-1 dosimetry in patients with B-lymphocytic malignancies. AB - BACKGROUND: Patients with B-cell non-Hodgkin's lymphoma (NHL) and chronic lymphocytic leukemia (CLL) frequently have splenomegaly, which has been reported to cause poor tumor targeting of radiolabeled antibodies. Consequently, patients with splenomegaly have been ineligible for some trials of radioimmunotherapy because of the assumption that they would not benefit. METHODS: Forty-nine patients with NHL and five with CLL received an initial dose of 131I-Lym-1 ranging from 740-8140 MBq. Six patients had prior splenectomy. The remaining 48 patients had spleen volumes ranging from 140-2830 ml determined using x-ray computed tomography. Medical Internal Radiation Dose Committee formalism was used to determine dosimetry, and spleen volume was used to adjust the S value for the spleen of each patient. RESULTS: Spleen radiation dose decreased as spleen volume increased, although there was a positive correlation (r = 0.75) between spleen volume and spleen cumulated activity. There was no clear relationship between spleen volume and tumor radiation dose, although tumor radiation doses were low in five patients whose spleen volumes were greater than or equal to 970 ml. There was no apparent relationship between spleen volume and therapeutic response to 131I-Lym-1. Two of five patients whose spleen volumes were greater than or equal to 970 ml responded despite low tumor radiation doses, whereas two of six patients with prior splenectomy did not respond. CONCLUSIONS: The results of this study provide no clear evidence that patients with splenomegaly should be excluded from radioimmunotherapy trials because of the assumption that they will not benefit. Splenomegaly was associated with decreased radiation dose to the spleen, and to tumors only for extraordinarily large spleens. PMID- 9406711 TI - Dosimetric comparison of bolus and continuous injections of CC49 monoclonal antibody in a colon cancer xenograft model. AB - BACKGROUND: Improved understanding of dose and effective dose calculations may contribute to the optimization of fractionated radioimmunotherapy. METHODS: Comparison three-dimensional tumor dosimetry was performed on athymic nude mice bearing established LS174T human colon carcinoma xenografts. Mice were given bolus intraperitoneal injections of 300 microCi 131I-labeled CC49 monoclonal antibody once (Day 0) or three times (Days 0, 3, and 7) or continuous intraperitoneal infusion with miniosmotic pumps over 7 days. Serial section autoradiography was used to reconstruct tumor activity density distributions for Days 3, 4, 7, 10, and 11 (single injection); Days 3, 4, 7, 8, and 11 (3 injections); and Days 4, 7, 10, and 13 (pump). At least three tumors were reconstructed at each time point. Uptakes in blood and tumor were measured up to 14 days (single injection), 11 days (3 injections), or 16 days (pump) after injection. RESULTS: Average dose values calculated from total activity uptake data only (assuming no energy loss external to the tumor) yielded 102 Gy (single injection), 158 Gy (three injections), and 47 Gy (pump). Average doses using three-dimensional dose calculations were 88 Gy, 139 Gy, and 40 Gy, respectively. The nonuniformity of dose deposition affects treatment outcome, because cell loss is an exponential function of dose. Using the linear quadratic model with fractional cell survival to define an effective dose, D(eff) were calculated to be 20 Gy, 23 Gy, and 14 Gy, respectively. Cell proliferation affects outcome for variable dose-rate treatments. With cell proliferation parameters set to reproduce single-fraction 60Co recurrence results, D(eff) (for local control endpoint) were 8.9 Gy, 12.8 Gy, and 3.9 Gy, respectively. Three bolus injections compared with a single bolus injection were relatively less efficient in tumor uptake. However, three bolus injections resulted in a more uniform dose rate over a longer period, resulting in a 50% improvement in D(eff). The slower dose delivery for pump infusion resulted in a significantly lower D(eff), although dose-rate distributions were more uniform compared with the single bolus injection. CONCLUSIONS: Improvement in dose-rate nonuniformities was observed for fractionated and continuous radiolabeled monoclonal antibody injections. Fractionated injections produced superior dosimetric results compared with single bolus or continuous injections. PMID- 9406710 TI - Imaging for improved prediction of myelotoxicity after radioimmunotherapy. AB - BACKGROUND: The severity of myelotoxicity after radioimmunotherapy has been predicted from body and blood radiation doses to marrow. However, marrow radiation can be increased substantially if the marrow or skeleton contains the malignancy targeted by the radiolabeled monoclonal antibodies. A study of 29 patients treated with iodine-131 (131I)-Lym-1 showed that radiation doses to marrow from body and blood had little correlation with myelotoxicity. The purpose of the present study was to assess the significance of marrow targeting and other factors for prediction of myelotoxicity. METHODS: Injected radioactivity and nontargeted radiation doses to marrow were compared with peripheral blood cell counts after the first therapy dose of 131I-Lym-1 in 16 heavily pretreated patients with non-Hodgkin's lymphoma (NHL). Bone marrow biopsy, targeted marrow radiation doses, marrow image uptake scores, age, Karnofsky performance score (KPS), previous chemotherapy, and tumor burden were also compared with blood counts. RESULTS: Myelotoxicity was not predicted well by injected radioactivity, total body radiation, or body and blood radiation doses contributed to marrow (P > 0.1). Biopsy-proven bone marrow lymphoma also failed to predict myelotoxicity (P > 0.1). Thrombocytopenia and leukopenia were predicted well by targeted radiation dose to marrow (P < 0.05) obtained by 131I imaging. Similarly, marrow image scores predicted decreases in platelets and white blood cells (WBCs; P < 0.05). Prediction of myelotoxicity using marrow radiation dose methods was slightly improved when serum lactic dehydrogenase (LDH), age, KPS, and prior chemotherapy were included in the analysis (P < or = 0.01). CONCLUSIONS: Prediction of myelotoxicity was improved in this group of patients by assessment of the targeting component of marrow radiation and was better predicted and obtained more easily by semiquantitative marrow image scores. Further improvement in prediction was slight when other factors were considered. PMID- 9406712 TI - Enhancement of 67Cu-2IT-BAT-LYM-1 therapy in mice with human Burkitt's lymphoma (Raji) using interleukin-2. AB - BACKGROUND: Lymphomas have been shown to be responsive to 131I immunoconjugates in studies conducted in mice and patients. We have observed that copper 67 (67Cu) labeled Lym-1 remains in lymphomatous tissue longer than 131I-Lym-1 and, consequently, results in higher absorbed radiation doses to tumors. In addition, recombinant interleukin-2 (rIL-2) has been reported to increase tumor uptake of radiolabeled antibody. Therefore, we examined the efficacy of 67Cu-labeled Lym-1 and the ability of rIL-2 to enhance this efficacy in athymic mice implanted with Raji xenografts. METHODS: 6[p-(bromoacetamido) benzyl]-1,4,8,11 tetraazacyclotetradecane-N,N', N'',N'''-tetraacetic acid (BAT) was conjugated to Lym-1 via 2-iminothiolane (2IT) to prepare 2IT-BAT-Lym-1, which was labeled with 67Cu. Mice with Raji xenografts were treated with 335-500 microCi (12.4-18.0 MBq) of 67Cu-2IT-BAT-Lym-1 with or without 48,000-144,000 IU of rIL-2 once or were treated b.i.d. for 5 days beginning simultaneously with 67Cu-2IT-BAT-Lym-1. Mouse weight, blood counts, and mortality were monitored to assess toxicity, and tumor size was measured to assess efficacy. In addition, groups of mice were sacrificed to assess the biodistribution of 67Cu-2IT-BAT-Lym-1 with and without rIL-2. RESULTS: In mice treated with 335 microCi of 67Cu-2IT-BAT-Lym-1 alone, 28% of tumors were cured. When 48,000 IU of rIL-2 were added, 50% were cured. The overall response-rate was 50% for both regimens. In mice treated with 400 microCi of 67Cu-2IT-BAT-Lym-1 alone, 42% responded, all of which were cured. When 48,000 IU of rIL-2 were added, 77% of tumors responded, and 38% were cured. Larger or multiple doses of rIL-2 did not result in additional therapeutic enhancement. The tumor uptake and radiation dose after 67Cu-2IT-BAT-Lym-1 were about two times greater when a single dose of rIL-2 was added: This may be the basis for enhanced therapeutic efficacy. Mortality was not altered for 335 microCi or 400 microCi doses of 67Cu-2IT-BAT-Lym-1 by rIL-2 nor were other toxicity parameters. Mortality was increased at 500 microCi by the addition of rIL-2. CONCLUSIONS: 67Cu-2IT-BAT-Lym-1 provided a therapeutic and frequently curative dose of radiation to tumored mice at tolerated doses. The therapeutic effectiveness of 67Cu-2IT-BAT-Lym-1 may have been enhanced by rIL-2. PMID- 9406713 TI - Importance of temporal relationships in combined modality radioimmunotherapy of breast carcinoma. AB - BACKGROUND: The beneficial effects of radioimmunotherapy (RIT) may result from activation of molecular pathways that lead to programmed cell death (apoptosis). The influences of sequence and timing of 90Y-DOTA-peptide-ChL6 antibody (90Y ChL6) and anti-epidermal growth factor receptor antibody (ch225) or paclitaxel (Taxol; Bristol-Myers Squibb, Princeton, NJ) on efficacy and toxicity were examined. METHODS: Groups of human breast carcinoma (HBT 3477) tumored mice received paclitaxel (300 or 600 microg) or ch225 (70, 150, or 350 microg) at various intervals before or after 90Y-ChL6. Mortality, hematologic toxicity, weight loss, and therapeutic efficacy were evaluated. RESULTS: Mice receiving paclitaxel within 24 hours of 90Y-ChL6 had a 100% response rate; 48% were cured when paclitaxel was given 6 or 24 hours after 90Y-ChL6. When 150 microg ch225 was given 24 hours before 90Y-ChL6, the response and cure rates surpassed those of 90Y-ChL6 alone. Timing of administration was critical, with mortality rates as high as 80% in some groups receiving 350 microg ch225 and 90Y-ChL6. CONCLUSIONS: In this aggressive human breast carcinoma model, combined 90Y-ChL6 and paclitaxel had a high therapeutic index with many cures. Sequence of administration was critical in order for ch225 or paclitaxel, when combined with 90Y-ChL6, to enhance the response rate. PMID- 9406714 TI - Overcoming the nephrotoxicity of radiometal-labeled immunoconjugates: improved cancer therapy administered to a nude mouse model in relation to the internal radiation dosimetry. AB - BACKGROUND: Elevated renal uptake and extended retention of radiolabeled antibody fragments and peptides is a problem in the therapeutic application of such agents. However, cationic amino acids have been shown to reduce renal accretion. The aims of the current study were to evaluate whether this methodology would benefit therapy with yttrium 90 (90Y)-labeled antibody fragments (Fab, F(ab)2), to establish the relationship between radiation dosimetry and observed biologic effects, and to compare the antitumor efficacy of antibody fragments with that of whole immunoglobulin (Ig)G. METHODS: The maximum tolerated dose (MTD) and the dose-limiting organ toxicity of 90Y-labeled anti-carcinoembryonic antigen (CEA) MN-14 monoclonal antibodies (Fab, F(ab)2, and IgG) were determined in nude mice bearing GW-39 human colon carcinoma xenografts. The mice were treated with or without kidney protection by administration of D-lysine, with or without bone marrow transplantation (BMT), or with combinations of each. Toxicity and tumor growth were monitored at weekly intervals after radioimmunotherapy. Dosimetry was calculated from biodistribution studies using 88Y-labeled antibody. Three different dosimetric models were examined: 1) taking solely self-to-self doses into account, using S factors for 90Y in spheroids from 0.1 to 1 g; 2) correcting for cross-organ radiation; and 3) using actual mouse anatomy as represented by nuclear magnetic resonance imaging with a three-dimensional internal dosimetry package (3D-ID). RESULTS: The kidney was the first dose-limiting organ with the use of Fab fragments. Acute radiation nephritis occurred at injected activities > or = 325 microCi, and chronic nephrosis at doses > or = 250 microCi. Activities of 200 microCi were tolerated by 100% of the animals (i.e., the MTD). Application of lysine decreased the renal dose by approximately fivefold, facilitating a 25% increase in the MTD (to 250 microCi), because myelotoxicity became dose-limiting despite red marrow doses of less than 5 gray (Gy). By using BMT and lysine, the MTD could be doubled from 200 to 400 microCi, where no biochemical or histologic evidence of renal damage was observed (kidney dose, < or = 40 Gy). With injected activities of > or = 325 microCi without kidney protection, and with a hepatic self-to-self dose of only 4 Gy, rising liver enzymes were observed, which could be explained only by cross-organ radiation from radioactivity in the kidneys (in the immediate neighborhood of the right kidney up to > or = 150 Gy). The MTD of F(ab)2 fragments could be elevated only by a combination of BMT and lysine. With IgG, the bone marrow alone was dose-limiting. Tumor dosimetry correlated well with antitumor effects; Fab was more effective than F(ab)2, which was consistent with its more favorable dosimetry, and it may also be more effective than IgG due to its higher dose rate and more homogenous distribution. Dosimetry Model 1 was insufficient for predicting biologic effects. Model 2 seemed to be more accurate, accounting for interorgan crossfire. However, Model 3 showed an additional substantial contribution to the red bone marrow dose due to crossfire from the abdominal organs. CONCLUSIONS: These data show that radiation nephrotoxicity is an important effect of cancer therapy with radiometal-conjugated antibody fragments or peptides. However, this effect can be overcome successfully with the application of cationic amino acids, which substantially increase the anti-tumor efficacy of radiometal-labeled immunoconjugates. For understanding the biologic effects (e.g., liver toxicity) of 90Y in a mouse model, accounting for cross organ radiation is essential. Further studies with radiometal-conjugated monoclonal antibody fragments and peptides are necessary to determine the MTD, dose-limiting organs, antitumor effectiveness, and nephroprotective effects of cationic amino acids in humans. PMID- 9406715 TI - Evaluation of cellular uptake, tumor retention, radiation response, and tumor pathophysiology in experimental solid tumors after an intratumoral infusion of colloidal 32P. AB - BACKGROUND: Order et al. successfully deposited > 85% of radionuclides in pancreatic carcinoma patients using an intratumoral (i. t.) infusion of macroaggregated albumin (MAA) prior to the i. t. infusion of colloidal 32P (32P CP), apparently due to MAA-induced transient blockade. However, no studies regarding physiologic response and the inhibitory tumor growth of solid tumors after treatment with 32P-CP with or without MAA have been performed. METHODS: First, the authors determined the cellular uptake of 32P-CP in 10 cell lines. They then evaluated the relationship between tumor retention and radiation response in vivo. Third, they evaluated the changes in physiologic parameters such as tumor interstitial fluid pressure (TIFP) and tumor blood flow (TBF) after an i. t. infusion of MAA. RESULTS: In the plateau growth phase of both H4IIE and LS174T tumors, maximal uptake occurred at approximately 100 minutes after treatment with 10 microCi of 32P-CP, but uptake in LS174T was approximately 2 times higher than that in H4IIE. In animal experiments, 32P-CP alone significantly inhibited the tumor growth of H4IIE. However, additional i. t. infusion of MAA did not improve the growth delay induced by 32P-CP, mainly due to insignificant differences in retention of radioactivity when using 32P-CP with or without MAA. It was speculated that when the outflow of the tumor vasculature was obstructed by clamping (or an i. t. infusion of MAA), it would reduce TBF and increase TIFP. However, neither increased TIFP nor decreased TBF was observed when MAA was given i. t. to tumors. CONCLUSIONS: The authors concluded that an MAA-induced transient blockade of tumor vasculature after an i. t. infusion of MAA did not occur in experimental solid tumors. PMID- 9406716 TI - Therapy for colon carcinoma xenografts with bispecific antibody-targeted, iodine 131-labeled bivalent hapten. AB - BACKGROUND: One of the main limitations of radioimmunotherapy (RIT) is the secondary toxicity related to the poor therapeutic indices achieved with labeled whole immunoglobulin (Ig)G or F(ab')2 fragments. To overcome this problem, we have developed a two-step targeting method, which we refer to as the Affinity Enhancement System (AES), using a radiolabeled bivalent hapten and a bispecific antibody recognizing the hapten and a target cell antigen. This method has been applied successfully to immunoscintigraphy in carcinoembryonic antigen (CEA) expressing carcinoma patients and increased tumor to normal tissue uptake ratios have been achieved. The aim of the current study was to evaluate the application of AES to RIT of CEA-expressing solid tumors in an animal model. METHODS: Nude mice grafted with LS174T human colorectal carcinoma were treated either with 111 megabecquerels (MBq) of iodine-131 labeled bivalent diethylenetriamine pentaacetic acid (DTPA) hapten 20 hours after pretargeting by anti-CEA x anti DTPA-indium bispecific antibody or 12 MBq of iodine-131 labeled anti-CEA IgG. RESULTS: Treatment with the IgG induced only a growth delay of 53 +/- 5 days but all tumors progressed. Treatment with the AES was highly efficient because tumor growth inhibition was achieved over 150 days. Hematologic and overall toxicity of both treatments were equivalent. CONCLUSIONS: The long term tumor regression consecutive to AES RIT represents a very significant improvement over the use of directly labeled IgG. Toxicity consecutive to AES or IgG RIT were similar despite an administered activity nearly ten times higher with the AES. However, given the efficacy of the AES treatment, a lower dose may afford lower toxicity and significant antitumor effect. PMID- 9406717 TI - Defining the optimal spacing between repeat radioantibody doses in experimental models: is there an accurate measurement for hematopoietic recovery? AB - BACKGROUND: Single doses of radioantibody are effective at treating single cells or small clusters of cancer cells. However, large tumor masses require either multiple doses of radioantibody or a multimodal approach to therapy using two or more therapeutic agents. Timing of the second dose in a multiple cycle scheme or the second treatment in a multimodal protocol will depend on recovery from toxicity associated with the first treatment. METHODS: BALB/c mice were dosed with a maximal tolerated dose (MTD) of I-131-MN-14 anti-carcinoembryonic antigen immunoglobulin G (IgG) (250 microCi) or F(ab')2 (1.2 mCi). Mice were redosed with the MTD at one of four time points, either Day 28, 35, 42, or 49 after IgG or Day 14, 21, 28, or 35 after F(ab')2. Survival was monitored to determine the earliest time to redose without lethality. Several studies were then performed to identify an accurate measure of true myelorecovery. Mice were bled retroorbitally on the day of the first dose and at weekly intervals thereafter. Total peripheral white blood cell counts, granulocyte counts, and lymphocyte counts were determined for each animal. GR-1hi expression (percentage of positive cells) and mean channel florescence were determined by FACScan analysis of a blood sample incubated with fluorescein isothiocyanate-anti-mouse Ly-6G (GR-1). In other studies, two mice were killed weekly from a group treated with a single MTD of radioantibody. The weights of their spleens and thymus glands were determined. At that time, femoral marrow was collected from these animals and plated in Methocult M3430 methylcellulose medium (Stemcell Technologies, Vancouver, Canada), and total colony-forming cells in culture were determined. Another population of mice was used to assess normal tissue metabolic activity following radioantibody therapy by quantitating the 4-hour utilization of I-125-dUrd. RESULTS: The ability to redose mice with a second MTD of 1-131-IgG or F(ab')2 required 49 days and 35 days, respectively. Granulocyte and lymphocyte counts did not accurately predict myelorecovery from the first dose. Hematopoietic tissue weight, tissue metabolic activity, and marrow colony forming cells all suggested that redosing was possible 1-2 weeks before it could actually be done without lethality. Percent of cells expressing GR-1hi (>60%) and absolute numbers of GR-1hi cells (>1400 cells/mm3) suggested myelorecovery in most animals. A greater degree of accuracy was achieved when trends in GR-1hi expression were noted over 2 or more weeks (i.e., the absolute amount of GR-1hi had to exceed levels in untreated mice, as evidence that the hyperproliferative phase of recovery was complete). CONCLUSIONS: The only approach that accurately predicted the ability to retreat with myelosuppressive therapy without risk of lethality was an increase in GR-1hi positive cells above untreated levels. Other approaches are currently being investigated, including the expression of proliferation antigens (e.g., proliferating cell nuclear antigen and Ki-67) in both murine and human samples and differentiation antigens (CD33 and CD34) in humans. PMID- 9406718 TI - Advantage of yttrium-90-labeled over iodine-131-labeled monoclonal antibodies in the treatment of a human lung carcinoma xenograft. AB - BACKGROUND: The purpose of this investigation was to compare the therapeutic effectiveness of yttrium-90 (90Y)-labeled monoclonal antibodies (MoAbs) with iodine-131 (131I)-labeled MoAbs delivered to human tumor xenografts at their maximum tolerated doses (MTD). METHODS: Nude mice bearing size-matched human lung adenocarcinoma xenografts (Calu-3) with mean tumor dimensions of approximately 0.8 cm received intravenous injections of the MTD of either 90Y- or 131I-labeled MoAbs. The mice received 125 microCi of 90Y-RS11 or 90Y-RS7, or 300 microCi of 131I-RS11 or 131I-RS7. Tumor size was measured weekly. Body weight and blood counts were monitored to measure toxicity. RESULTS: The calculated average radiation doses delivered to tumors in mice treated with 90Y-RS11 and 90Y-RS7 were 3578 and 3787 centigray (cGy), respectively, versus 1264 and 1368 cGy for 131I-RS11 and 131I-RS7, respectively. These values were calculated taking into consideration the tumor size-dependent absorbed fractions for the beta particles of both radioisotopes. The calculated radiation doses absorbed in the tumor correlated with the tumor responses observed. Mean tumor volume decreases at nadir were 66% and 87% after 90Y-RS11 and 90Y-RS7 therapy, respectively, with 15 of 30 mice treated with either MoAb having complete regression of their tumors. In contrast, only mean stable disease was observed with the 131I MoAb treatments. CONCLUSIONS: The results demonstrate the superiority of 90Y-labeled MoAbs over conventionally labeled 131I-labeled MoAbs for therapy in this model. The fact that a substantial percentage of complete remissions was achieved with 90Y labeled MoAbs suggests a considerable radiosensitivity of the tumor studied, and supports the suitability of choosing this tumor type as an attractive target for future clinical radioimmunotherapy. PMID- 9406719 TI - Preclinical analysis of radiolabeled anti-GD2 immunoglobulin G. AB - BACKGROUND: Unlabeled murine monoclonal anti-GD2 immunoglobulin (Ig)G (14G2a) reactive with nervous system diganglioside and neuroblastoma, melanoma, and small cell lung carcinoma produces tumor regression. However, serious acute abdominal pain, paresthesia, hypotension and hypertension, syndrome of inappropriate secretion of antidiuretic hormone (SIADH), and occasional motor weakness occur. Studies in preclinical animal models can elucidate the mechanism of the observed neurotoxicity and lead to anti-GD2 antibody treatment with a higher therapeutic ratio. METHODS: One mg of 14G2a or control IgG was labeled with 1-2 mCi of indium 111 and administered intravenously to beagles (n = 8). In 2 dogs, additional high dose (200 mg) unlabeled 14G2a was given over 5 days. Whole body gamma camera images and SPECT scans were obtained repeatedly over 7 days. On Day 7, sciatic nerve conduction studies were performed, and after euthanasia radioactivity was determined in major organs. RESULTS: Unlabeled high dose 14G2a administered to mice, rats, or rabbits did not cause neurotoxicity within 3 weeks. GD2 antigens were shown by immunochemistry to be present in brain and peripheral nerve tissues of rodents and beagles. After in vivo administration of radiolabeled 14G2a, canine lymph nodes showed specific uptake, but only minimal radioactivity was found in the nervous system. Dogs that received additional high dose unlabeled 14G2a showed much higher lymph node uptake and follicular lymph node hyperplasia. Low motor response amplitudes on nerve conduction studies were noted. CONCLUSIONS: A radioisotope label on IgG and its visualization in a large series of animal models indicate that a low protein dose of anti-GD2 IgG will not cause neurologic side effects in patients. High protein dose anti-GD2 IgG may enhance antineoplastic effects and contribute to neurotoxicity through stimulation of normal lymphocytes with subsequent release of cytokines. PMID- 9406720 TI - Biodistribution of iodine-125 labeled monoclonal antibody/interleukin-2 immunoconjugate in athymic mice bearing human tumor xenografts. AB - BACKGROUND: Some vasoactive drugs have been studied in the hope of altering the vascular permeability and/or blood of tumors to enhance monoclonal antibody (MoAb) uptake. The pretreatment of interleukin-2 (IL-2), one of the vasoactive reagents, produced a generalized vascular permeability, but its function was not tumor specific. Conversely, MoAb/IL-2 immunoconjugates were developed that selectively alter the vasopermeability of tumors. In the current study the authors evaluated whether radiolabeled anti-carcinoembryonic antigen (CEA) MoAb, ZCE025/IL-2 immunoconjugate, specifically can enhance delivery of iodine-125 (I 125) to tumor sites. METHODS: ZCE025 was conjugated with IL-2, and the conjugate then labeled with I-125. Biodistribution studies were performed in athymic mice bearing CEA-producing human tumor (MKN45) xenografts. Mice were injected with I 125-ZCE025/IL-2 conjugate, and I-125 activities in the organs, including blood and tumor, were investigated at 1, 3, and 5 days after injection. Vascular permeability of the organs, including tumors, also was studied by using I-131 labeled mouse serum albumin in three groups. RESULTS: I-125 labeled ZCE025/IL-2 conjugate destroyed its lymphokine-activated killer cell (LAK) activation, but retained a minimum of 75% of the antibody binding reactivity. Biodistribution of I-125-ZCE025/IL-2 conjugate showed increased uptake of I-125 in tumor by a factor of 1.5, 4.2, and 4.1 compared with I-125-ZCE025 on Days 1, 3, and 5, respectively. In contrast, I-125 distribution was not enhanced in any organs except the tumor. Vascular permeability studies demonstrated that the physiologic effect of IL-2 was tumor specific in the mice that received the I-125-ZCE025/IL-2 conjugate. CONCLUSIONS: These studies indicate that the administration of radiolabeled MoAb/IL-2 double conjugate may enhance the therapeutic potential of radiolabeled MoAb without any pretreatment with IL-2 or repeated injection of MoAb. PMID- 9406721 TI - Can occult metastases be treated by radioimmunotherapy? AB - BACKGROUND: Tumor lesions in the millimeter (mm) range may escape detection with nuclear medicine imaging methods (including single photon emission computed tomography [SPECT]) using radiolabeled monoclonal antibodies (MoAbs). We hypothesized that these lesions still could receive a potentially therapeutic radiation absorbed dose, and therefore should be treated, despite the lack of detection. METHODS: To simulate this situation, 2-mm beads (0.004 mL) containing approximately 1.15 microCi of iodine-131 (131I) were used. The beads were placed centrally in a 1200-mL liver phantom containing approximately 3 mCi of 131I. The resultant activity concentration on the beads was approximately 288 microCi/mL compared with approximately 2.5 microCi/mL in the phantom, corresponding to a maximum tumor uptake of approximately 0.3% injected dose per gram (%ID/g) if 100 mCi of 131I-labeled immunoglobulin G were administered. The phantom, containing the beads, was imaged by both planar and SPECT techniques at hypothetical Day 1 (time of maximum tumor uptake) and at hypothetical Day 7 to examine the improved target-to-nontarget ratio over time. In addition to imaging the beads, the radiation absorbed dose to the simulated lesions from the beta component emissions of 131I was calculated using absorbed fractions based on Berger's point kernels. RESULTS: Regardless of the conditions used, the beads could not be observed by either planar or SPECT imaging. However, the radiation-absorbed dose to the simulated lesion was calculated to be as high as approximately 6200 centigray (cGy), with an average dose rate of approximately 89.5 cGy/hour. CONCLUSIONS: This simulation demonstrates that a relatively high absorbed dose and dose rate can be delivered to mm-sized lesions not observed by conventional nuclear imaging methods, and that these lesions should be considered for radioimmunotherapy with 1311 MoAbs. However, for micrometastases of <1 mm, other radionuclides with shorter path length beta particles than 131I, Auger electrons, or alpha particles should be considered. PMID- 9406722 TI - Generation of a high-producing clone of a humanized anti-B-cell lymphoma monoclonal antibody (hLL2). AB - BACKGROUND: LL2 is a murine immunoglobulin (Ig)G2a-kappa anti-B-cell monoclonal antibody with proven targeting and therapeutic efficacy in the management of non Hodgkin's lymphoma (NHL). The authors had previously generated a humanized LL2 (hLL2) that demonstrated binding properties identical to those of LL2. Nevertheless, the productivity of the cell line was insufficient for large-scale production of the antibody for clinical studies. Therefore, the authors chose an amplifiable system for the generation of hLL2. METHODS: The hLL2 sequences were ligated into the expression vector pdHL2, which has a dhfr amplifiable gene, and were incorporated into the SP2/0 cells by electroporation. A methotrexate (MTX) resistant clone producing hLL2 was identified. Stepwise increases in MTX concentrations, from 0.1 to 5 microM, and subcloning of the cells by limiting dilution were performed. RESULTS: By amplifying the dhfr and hLL2 genes with stepwise increases in the MTX concentration, the antibody production was enhanced from its original 1.4 to 70 +/- 5 mg per liter of culture media. Subsequent subcloning further improved the productivity. Immunoreactivity of the antibody was conserved, as proven by enzyme-linked immunosorbent assay and cell-binding assays. By isoelectrofocusing, the isoelectric point (pI) of the antibody was measured at approximately 9.6. The productivity of the clone was not affected by culture conditions or storage of the cells in liquid nitrogen. CONCLUSIONS: By means of gene amplification, the authors have generated a high-producing hLL2-IgG clone suitable for production of the quantity of antibody necessary for clinical diagnostic and therapeutic trials of NHL patients. PMID- 9406723 TI - Chimerization of Mu-9: a colon-specific antigen-p antibody reactive with gastrointestinal carcinomas. AB - BACKGROUND: Mu-9 is a murine monoclonal antibody that is directed against affinity-purified colon-specific antigen-p (CSAp). CSAp is a tumor-associated antigen that is present in 60% of colorectal carcinomas. Preclinical and clinical studies have shown Mu-9 to have excellent targeting abilities. However, as administration of the murine immunoglobulin G (IgG) provoked a human anti-mouse antibody response, chimerization of Mu-9 is warranted for decreasing immunogenicity. METHODS: Polymerase chain reaction and cDNA library screening methods were used for the cloning of Mu-9 heavy and light chain variable regions for the construction of chimeric Mu-9. RESULTS: The functional chimeric Mu-9 antibody binds to the CSAp antigen in the GW-39 extracts. It has immunochemical properties similar to that of murine Mu-9. CONCLUSIONS: The V-region sequence information will be used for design of humanized Mu-9, which will be evaluated for targeting gastrointestinal carcinomas. PMID- 9406724 TI - Pretargeting: general principles; October 10-12, 1996. AB - BACKGROUND: Human imaging studies show that maximum human tumor concentrations of monoclonal antibody (MoAb) are achieved in 1 day, but several days are required for background reduction and sensitive radioimmunoscintigraphy of tumors. With therapeutic radionuclides such as yttrium-90 (90Y), this long biologic half-life imposes a high radiation burden on sensitive normal tissues from the large amount of unlocalized radioactivity. Normal tissue toxicity, especially to the bone marrow, has been the major limiting factor in the application of radioimmunotherapy to solid tumors. Pretargeting techniques provide an alternative method with which to obtain high selective tumor uptake of 90Y with simultaneous minimization of nontarget tissue background. METHODS: Current MoAb techniques employ either the biotin/avidin or the hapten/antibody system. DNA/DNA and prodrug/enzyme systems also have been used and many other ligand/receptor systems are possible. All the methods depend on a long circulating conjugate to obtain high target uptake with a diffusible, rapidly excreted effector molecule. RESULTS: Fast in, slow out tumor kinetics, ideal for therapy, have been achieved. In one mouse tumor system the biologic half-life of a bivalent 88Y JANUS tetraazacyclodidecanetetraacetic acid hapten, measured over 5 days, was approximately 24 hours. The therapeutic ratio, obtained from the integrated tumor and blood concentrations over 5 days, was approximately 20/1 compared with 2-3/1 with directly labeled MoAb. The total injected dose remaining in the mouse at 24 hours was 5.5%, 23% of which was in the tumor. CONCLUSIONS: These result suggest that it may be possible to deliver tumoricidal radiation doses with 90Y using pretargeting techniques without severe normal bone marrow irradiation. PMID- 9406725 TI - Dosimetry of fractionated experimental radioimmunotargeting with idiotypic and anti-idiotypic anticytokeratin antibodies. AB - BACKGROUND: Repeated injections of iodine-125 (125I)-labeled tumor targeting anticytokeratin monoclonal antibody (TS1) and a nonlabeled antiidiotypic monoclonal antibody against TS1 (alphaTS1) were compared with a single injection of the radiolabeled TS1 in experimental radioimmunotargeting. Anti-TS1 was used to remove nontargeting TS1. METHODS: Nude mice were inoculated with HeLa Hep2 cells. The animals in Group A received a single injection of 13 MBq 125I-labeled TS1. The animals in Group B received four injections of 125I-labeled TS1 (8-13 MBq) followed by alphaTS1 24 hours later, at 2-week intervals. The mean absorbed doses were calculated according to the Medical Internal Radiation Dose Committee criteria based on repetitive radioimmunoscintigraphies during an observation period of 59 days. RESULTS: A 11 gray (Gy) mean dose to the tumor and 2 Gy to the whole body was achieved in Group A. Mean peak tumor uptake of 5% of the injected dose (ID), corresponding to 14% ID/g, was observed on Day 17 after a single injection of the labeled monoclonal antibody. A mean peak tumor uptake of the same order of magnitude was seen in Group B. An absolute increase in the tumor uptake was observed in Group B during the entire observation period. The mean absorbed dose to the tumors was 11 Gy at the end of the observation period, whereas the whole body dose was only 2.5 Gy in Group B. Autoradiography of the tumors at the end of the observation period confirmed an intensive heterogeneous accumulation of activity in the entire tumor. CONCLUSIONS: The fractionated strategy can contribute to a significant accumulation of radiolabeled TS1 in the tumors. Furthermore, the use of alphaTS1 makes it possible to increase the tumor to-nontumor dose ratio and maintain a prolonged high activity accumulation in the tumor. PMID- 9406726 TI - Experimental radioimmunotargeting combining nonlabeled, iodine-125-labeled, and anti-idiotypic anticytokeratin monoclonal antibodies: a dosimetric evaluation. AB - BACKGROUND: Preinjection of a nonlabeled tumor targeting anticytokeratin monoclonal antibody (TS1) and postinjection of an anti-idiotypic anticytokeratin monoclonal antibody (alphaTS1) were evaluated separately and in combination to investigate their effects on the accumulation of iodine-125 (125I)-labeled TS1 in experimental radioimmunotargeting. TS1 targets deposited extracellular cytokeratin 8 from necrotic tumor cells. METHODS: Nude mice were inoculated with HeLa Hep 2 cells. Four different groups were followed with 504 repetitive quantitative radioimmunoscintigraphic recordings during a 78-day observation period. The absorbed doses were calculated according to criteria of the Medical International Radiation Dose Committee. RESULTS: As much as 2% of the injected dose (ID) of 125I-labeled TS1 accumulated in the tumor, and the peak tumor uptake was recorded as late as Day 30 after the injection of 125I-labeled TS1. Anti-TS1 caused a rapid decrease in the whole body activity. The highest tumor-to-nontumor activity ratios were obtained when a pre-injection of nonlabeled TS1 was combined with a postinjection of alphaTS1. The mean absorbed dose in tumor per unit activity administered was 0.44 gray/megabecquerel (Gy/MBq) and in nontumor tissues 0.15 Gy/MBq after a single injection of 125I-TS1. The efficacy was 0.34 Gy/MBq in tumor and 0.1 Gy/MBq in nontumor tissues after a combination of preinjection of nonlabeled TS1 and postinjection of nonlabeled alphaTS1. This indicates a 20% increase in tumor doses compared with a single injection of labeled TS1. CONCLUSIONS: This study confirms an extensive accumulation of TS1 in the tumor, with peak values as late as 30 days after injection of labeled TS1. Furthermore, both preinjection of nonlabeled TS1 and postinjection of alphaTS1 can improve radioimmunotargeting. PMID- 9406727 TI - Pretargeting using peptide nucleic acid. AB - BACKGROUND: Pretargeting studies in animals and humans have usually involved (strept)avidin and biotin. Depending on the particular strategy, endogenous biotin can adversely influence localization when these molecules are used. METHODS: As an alternative to (strept)avidin and biotin, we have explored the use of a single-stranded peptide nucleic acid (PNA) bound to a protein administered first and followed by the complementary single-stranded PNA radiolabeled with 99mTc. Target localization of the PNA-bound protein in a mouse infection and a mouse tumor model occurred by passive diffusion while the radiolabeled complementary PNA localized by in vivo hybridization. The PNA-streptavidin was prepared by adding biotin-conjugated PNA to streptavidin; the complementary PNA, derivatized with a primary amine, was conjugated with acetyl S-protected NHS-MAG3 bifunctional chelator and radiolabeled with 99mTc. RESULTS: In both the infection and tumor mouse models, increased localization of radiolabel was achieved in animals receiving both injectates compared with control animals receiving only the radiolabeled PNA. In the infection model, the infected to normal thigh radioactivity ratio was 3.5 for the study animals compared with 1.7 for control animals (P = 0.0001). In the tumor model, these values were 1.7 versus 1.2 (P = 0.003). CONCLUSIONS: We conclude that PNA may be considered an alternative to (strept)avidin and biotin for pretargeting studies. PMID- 9406728 TI - Increased survival associated with radiolabeled Lym-1 therapy for non-Hodgkin's lymphoma and chronic lymphocytic leukemia. AB - BACKGROUND: Because most patients with advanced non-Hodgkin's lymphoma (NHL) and chronic lymphocytic leukemia (CLL) respond to radioimmunotherapy (RIT), the study was designed to evaluate the relationship between response and survival in patients treated with radiolabeled Lym-1. METHODS: Fifty-seven patients with NHL (52) or CLL (5) were treated with radiolabeled Lym-1 antibody between 1985 and 1994. Influence of response to treatment on survival was examined directly and was also adjusted for other factors previously found to be significant predictors of survival. A multivariate model that was based on baseline Karnofsky performance status (KPS) and serum lactic dehydrogenase (LDH) predicted response and survival and was used to define risk groups. Proportional-hazards, Kaplan Meier, and Landmark models were used to evaluate parameters for their ability to predict outcome. RESULTS: By using a proportional-hazards model with response as a time-dependent variable, overall response (P < 0.001) and complete response (P = 0.006) were predictive of increased survival in univariate analyses. Overall response continued to be significant (P = 0.02) in multivariate analyses, even when risk groups that also predicted survival were included. Median survival of responders was 84 weeks, whereas that of nonresponders was 22 weeks when the Landmark method, based on response status at 16 weeks from start of therapy, was used to generate survival curves. CONCLUSIONS: Response to treatment with radiolabeled Lym-1 was associated with increased survival, even when adjusted for baseline clinical parameters that also predicted for survival. The results provide the first unbiased evidence for survival effects of RIT in patients. PMID- 9406730 TI - Recurrence of Hodgkin's disease after indium-111 and yttrium-90 labeled antiferritin administration. AB - BACKGROUND: Indium-111 labeled antiferritin targets 95% of all Hodgkin's disease lesions with a diameter of 1 cm or more. Subsequent treatment with yttrium-90 labeled antiferritin secures a high response rate in patients with recurrent Hodgkin's disease. METHODS: A total of 87 patients were entered on one of three different yttrium-90 labeled antiferritin protocols. Recurrences after yttrium-90 treatment were analyzed. Nine patients were retreated with involved external beam radiation fields, selected with the help of indium-111 labeled antiferritin. RESULTS: In single-agent yttrium-90 antiferritin studies, a response rate of more than 60% was found, with an average response duration of 6 months. One-third of the patients had recurrences in previously uninvolved areas. Repeat indium antiferritin scintigraphy allowed for the selection of new radiation fields for recurrences. In-field disease control was obtained for a median of 8 months, but new recurrences in new areas occurred. Chemotherapy or radiation therapy given immediately before antiferritin decreased tumor targeting with indium-111 labeled antiferritin. CONCLUSIONS: Recurrences after radiolabeled antiferritin treatment are not due to radioresistant Hodgkin's disease. In contrast, Hodgkin's disease less than 1 cm in diameter is not targeted and not controlled by radiolabeled antiferritin. New multimodality regimens with a higher therapeutic ratio are needed for treatment of Hodgkin's disease with curative intent. Radiolabeled antiferritin can be incorporated in such regimens to secure better control of bulky Hodgkin's disease (>1 cm in diameter), but it should be given before chemotherapy or radiation therapy. PMID- 9406729 TI - Escalating protein doses of chimeric monoclonal antibody MOv18 immunoglobulin G in ovarian carcinoma patients: a phase I study. AB - BACKGROUND: Successful first-line treatment of ovarian cancer does not incite cure. Recurrent disease often shows an increased resistance to chemotherapeutic agents. Therefore, other treatment modalities, like (radio)immunotherapy using tumor-associated monoclonal antibodies, should be considered. Chimeric MOv18 (c MOv18) localizes well in ovarian carcinoma tissue. We investigated the safety of a single intravenous (i.v.) infusion of increasing doses of c-MOv18 in ovarian carcinoma patients. METHODS: Fifteen patients received c-MOv18 (from 5 mg to 75 mg). Safety was determined by recording vital signs; by hematological, biochemical, and urinary analyses; and by the human-antichimeric antibody (HACA) response. Five patients received c-MOv18 labeled with a tracer dose of iodine-131 to analyze the pharmacokinetics and biodistribution in blood, urine, and tissue biopsies at surgery. RESULTS: Administration of c-MOv18 IgG was uneventful. No significant changes in hematological, biochemical, or urine profiles were noted at any time postinjection (p.i). Starting with a dose of 50 mg, all patients experienced side effects, like fever, headache, and nausea/vomiting, maximally Grade II (World Health Organization toxicity scale). No HACA response was found up to 12 weeks p.i. The mean elimination half-life after infusion of 30-75 mg c MOv18 was significantly higher compared with infusion of 1 mg. Absolute amount of c-MOv18 in carcinoma tissue increased with increasing c-MOv18 doses. CONCLUSIONS: Intravenous administration of c-MOv18 IgG in a dose up to 75 mg is safe, inducing only minor side effects at doses of 50 mg or higher. In view of the characteristics of c-MOv18, this antibody might be applicable as an unmodified antibody or as an immunoconjugate in the treatment of ovarian carcinomas. PMID- 9406731 TI - Peripheral blood stem cell mobilization for hematopoietic support of radioimmunotherapy in patients with breast carcinoma. AB - BACKGROUND: In clinical trials of radiolabeled monoclonal antibodies targeting metastatic breast carcinoma, myelosuppression has been the initial dose-limiting toxicity. We previously have shown that mobilized peripheral blood stem cell transfusions ameliorate this toxicity of radioimmunotherapy (RIT). Because of the difficulty we experienced harvesting adequate numbers of precursor cells from some of these heavily pretreated patients, we have compared the mobilization results and clinical histories of the metastatic breast carcinoma patients referred for RIT with those of metastatic breast carcinoma patients referred for high dose chemotherapy (HDCT) with transplantation. METHODS: Mobilization of stem cells into the peripheral blood was accomplished using granulocyte-colony stimulating factor with or without chemotherapy. Granulocyte macrophage colony forming assays (CFU-GM) and flow cytometric analysis for CD34 positive cells were used to evaluate the number of hematopoietic precursors mobilized and collected with each apheresis procedure. Clinical characteristics, including prior chemotherapy and external beam radiotherapy, tumor bulk, and performance status, were determined by chart review. RESULTS: Significantly fewer hematopoietic precursors (both CD34 positive cells and CFU-GM) were harvested per procedure from the six RIT patients compared with a group of six patients with metastatic breast carcinoma who had stem cells harvested prior to HDCT (P = 0.002). There was no significant difference between the groups with regard to age or prior radiotherapy. All the RIT patients had received more chemotherapy, had a less favorable performance status (1 vs. 0), and had measurable tumor, whereas all the HDCT patients had minimal residual disease. CONCLUSIONS: Patients enrolled in RIT studies had lower stem cell yields than metastatic breast carcinoma patients scheduled to receive HDCT with stem cell transplantation. Poor mobilization is unlikely to be due to the mobilizing regimen alone and may be related to the intensity of prior therapy and/or tumor bulk. Mobilizing adequate stem cells for multiple treatment cycles from patients on Phase I/II RIT trials may require new, more effective mobilizing regimens, but referral of patients earlier in their disease course, prior to chronic bone marrow damage and disease progression, is recommended strongly. PMID- 9406732 TI - Local application of radiolabeled monoclonal antibodies in the treatment of high grade malignant gliomas: a six-year clinical experience. AB - BACKGROUND: Infusion of radiolabeled monoclonal antibodies (MAbs) directly into a tumor or into the site of disease after surgery concentrates a high quantity of antibody and radioisotope in the neoplastic tissue. The strong irradiation delivered by this method can result in control of high grade malignant gliomas. METHODS: Antitenascin MAbs BC-2 and BC-4 labeled with 131I (mean dose, 1998 MBq) were injected into 105 patients with malignant glioma by means of an in-dwelling catheter. Multiple courses (up to six) were given. The patients underwent MAb treatment after their tumors were minimized by surgery, radiotherapy, and, in recurrent lesions, a second operation. Data is presented in this article for 62 evaluable patients with high grade malignant gliomas (58 glioblastomas and 4 anaplastic astrocytomas), of which 31 were newly diagnosed tumors and 31 were recurrent lesions. In 40 cases the disease was minimal at the time of MAb injection, and in 22 cases a macroscopic remnant was present. RESULTS: There were very few adverse effects, all of which were minor. The treatment yielded a significant extension of patients' median survival (23 months) and of the disease free time to relapse (12 months). Favorable objective responses were recorded as follows: 9 partial responses, 3 complete responses, and 20 with no evidence of disease. A response rate of 51.6% was calculated for all assessable patients. The most important factor in obtaining beneficial outcomes was limited extension of the neoplasm at the time of therapy. CONCLUSIONS: In selected patients, locoregional radioimmunotherapy can be included in a multimodal strategy to control high grade malignant gliomas and produce favorable results. PMID- 9406733 TI - Objective responses after fractionated infusional brachytherapy of unresectable pancreatic adenocarcinomas. AB - BACKGROUND: The prognosis of unresectable pancreatic adenocarcinoma is poor. Therefore, the treatment potential of an intratumoral infusional brachytherapy using macroaggregated human albumin in combination with radioactive chromic phosphate [32P] was investigated in this group of patients. METHODS: Seventeen patients with unresectable tumors received intratumoral infusional brachytherapy. Treatment and assessment of response was performed with the aid of ultrasonography. RESULTS: Four patients had complete response with a duration ranging from 2-57 weeks and 5 patients had partial response with a duration ranging from 4-21 weeks, corresponding to an objective response of 53% (9 of 17 patients). Six of these patients were alive 33-57 weeks after treatment. Radiation necrosis was observed in 1 patient after a 19,000-gray cumulative radiation dose and a slight decrease in blood counts was observed in 2 patients. CONCLUSIONS: Intratumoral infusional brachytherapy using radioactive colloidal chromic phosphate has the potential to reduce inoperable pancreatic tumors with few side effects. PMID- 9406734 TI - Factors influencing hematologic toxicity of radioimmunotherapy with 131I-labeled anti-carcinoembryonic antigen antibodies. AB - BACKGROUND: Several investigators have reported a considerable variability in the observed hematologic toxicity after radioimmunotherapy (RAIT) with monoclonal antibodies (MoAb) given at similar amounts of radioactivity based on body surface area and/or similar radiation absorbed doses given to the red marrow. The authors investigated various factors potentially affecting hematologic toxicity after RAIT with 131I-labeled anti-carcinoembryonic antigen (CEA) MoAb to identify the statistically significant factors from those commonly perceived clinically to substantially contribute to this toxicity. METHODS: Ninety-nine patients who received 131I-labeled anti-CEA MoAb for the treatment of CEA-producing cancers were assessed for platelet and white blood cell toxicity based on the common Radiation Therapy Oncology Group criteria. Multivariate regression analysis was used to identify the statistically significant factors affecting toxicity among the following variables: red marrow dose, baseline platelet and white blood cell counts, bone and/or marrow metastases, prior chemotherapy or radiotherapy, timing of prior chemotherapy or radiotherapy in relationship to RAIT, type and number of prior chemotherapeutic regimens, age, sex, antibody form, and cancer type. RESULTS AND CONCLUSIONS: Red marrow dose, baseline platelet or white blood cell counts, multiple bone and/or marrow metastases, and chemotherapy 3-6 months before RAIT were the only four significant factors affecting hematologic toxicity according to multivariate analysis. The identification of bone and/or marrow metastases and recent chemotherapy as significant factors for hematologic toxicity could be important in the design of future clinical trials. PMID- 9406735 TI - Folate and cobalamin in psychiatric illness. AB - The linkage of cobalamin and folate deficiency to psychiatric illness has been studied and debated since these vitamins were first discovered in the 1940s. The clinical relevance of these deficiencies remains the subject of investigation and scholarly discussion. This article reviews case reports and studies derived from a MEDLINE search for English-language articles related to folate, cobalamin, and psychiatric illness. Emphasis is given to clinical research and recent developments. Preclinical evidence for direct effects of folate and cobalamin on brain functioning is compelling, and numerous associations of their deficiencies to psychiatric illness are evident. These vitamin deficiencies may typically present initially with psychiatric symptoms, but any direct causal relationship to specific neuropsychiatric illnesses are not well defined. The relationship of these vitamins in dementia is significant, but they may only rarely be a cause of truly reversible dementia. Folate deficiency appears most tightly connected with depressive disorders, and cobalamin deficiency with psychosis. Contrary to intuition, vitamin deficiencies appear to occur infrequently with eating disorders. Other diagnoses have been investigated much less extensively. The diagnosis and management of these deficiencies in the context of neuropsychiatric illness is still a matter of discussion. The quality of clinical research in this area is improving, but there are many unanswered questions that affect clinical practice. Clinicians should remain vigilant to the possibility of deficiencies of folate and cobalamin in diverse psychiatric populations. Normal hematological indices do not rule out the deficiencies. Further study is needed to refine the detection and clinical management of these vitamin deficiencies in psychiatric populations. PMID- 9406736 TI - Comorbidity of panic disorder with agoraphobia and specific phobia: relationship with the subtypes of specific phobia. AB - The study objectives were to determine comorbidity rates for various subtypes of specific phobia (SP) in a sample of patients with the principal diagnosis of panic disorder with agoraphobia (PDA) and to examine the possible etiologic relatedness of these SP subtypes to PDA. Ninety consecutive day clinic patients with PDA were administered the Structured Clinical Interview for DSM-III-R (SCID) modified for DSM-IV. The overall comorbidity rate for SP was 65.6%. The most frequent subtypes of SP were situational phobia and dental phobia, followed by natural environment phobia, phobia of funerals, cemeteries, dead bodies, and other death-related phenomena and objects (referred to as death-related phobia), and blood-injection-injury phobia. Except for death-related phobia, other subtypes of SP clearly tended to precede the onset of PDA, often by many years. The smallest difference between the age of onset for PDA and particular subtypes of SP (temporal distance) was found for death-related phobia, whereas the temporal difference was longer for situational phobia, hospital phobia, and blood injection-injury phobia. The frequency and temporal distance data suggest that death-related phobia may constitute a risk factor for developing PDA or that it is a prodrome of PDA, whereas situational phobia, hospital phobia, and blood injection-injury phobia appear to predispose to PDA to a lesser degree. Of the three broadly conceived groups of SP, mutilation phobias (which include death related phobia, hospital phobia, blood-injection-injury phobia, and dental phobia) appear most etiologically relevant for PDA, with the group of situational phobias (which also includes the natural environment subtype of SP) being less relevant, and animal phobias showing a negligible etiologic relatedness to PDA. PMID- 9406737 TI - The prevalence of high-level exercise in the eating disorders: etiological implications. AB - There is increasing evidence both from animal experimentation and from clinical field studies that physical activity can play a central role in the pathogenesis of some eating disorders. However, few studies have addressed the issue of prevalence or whether there are different rates of occurrence across diagnostic categories, and the estimates that do exist are not entirely satisfactory. The present study was designed to conduct a detailed examination of the physical activity history in patients with anorexia nervosa (AN) and bulimia nervosa (BN) both during and prior to the onset of their disorder. A sample of adult patients and a second sample of adolescent AN patients took part in the study. A series of chi-square analyses compared diagnostic groups on a number of variables related to sport/exercise behaviors both premorbidly and comorbidly. Data were obtained by means of a detailed structured interview with each patient. We found that a large proportion of eating disorder patients were exercising excessively during an acute phase of the disorder, overexercising is significantly more frequent among those with AN versus BN, and premorbid activity levels significantly predict excessive exercise comorbidity. These findings underscore the centrality of physical activity in the development and maintenance of some eating disorders. They also have important clinical implications in light of the large proportion of individuals who combine dieting and exercise in an attempt to lose weight, and the increasing recognition of the adverse effects of strenuous physical activity in malnourished individuals. PMID- 9406738 TI - Dissociative detachment relates to psychotic symptoms and personality decompensation. AB - Previous studies have addressed the prominence of psychotic symptoms in conjunction with multiple personality disorder (now dissociative identity disorder). The present study examines the relation between psychotic symptoms and a more pervasive form of dissociative disturbance, namely dissociative detachment. Two hundred sixty-six women in inpatient treatment for severe trauma related disorders completed the Dissociative Experiences Scale (DES), and 102 of these patients also completed the Millon Clinical Multiaxial Inventory (MCMI III). A factor analysis of the DES yielded two dimensions of dissociative detachment: detachment from one's own actions and detachment from the self and the environment. Each of these DES dimensions relates strongly to the thought disorder and schizotypal personality disorder scales of the MCMI-III. We propose that severe dissociative detachment, by virtue of loosening the moorings in inner and outer reality, is conducive to psychotic symptoms and personality decompensation. PMID- 9406739 TI - Age and cognitive impairment influence the performance of the General Health Questionnaire. AB - The General Health Questionnaire (GHQ) is a screening instrument designed to detect nonpsychotic psychiatric disorders. Its discriminating ability can be influenced by factors such as the presence of physical illness, comorbidity with other psychiatric disorders, and the presence of cognitive impairment, which are more frequent in the elderly. The present study examines the influence of age and cognitive impairment on the performance of the GHQ-12, and was performed in the course of a family study designed to evaluate the risks for dementia, depression, and geriatric depression in the relatives of elderly subjects with dementia of Alzheimer type or major depression. Four hundred subjects who had completed the GHQ-12 were included. Test performance was evaluated by receiver-operating characteristic (ROC) analysis. Our results indicate that (1) the GHQ-12 is applicable to elderly subjects (>65 years), (2) its performance is comparable in different age groups, (3) the cutoff value for case identification is higher in the elderly (3/4) compared with younger individuals (1/2) and (4) mild cognitive impairment does not influence the good performance of the GHQ-12 in elderly subjects. In conclusion, our study verifies the usefulness of the GHQ-12 as an instrument to identify states of depression; this applies also to subjects with mild intellectual impairment. To optimize the discriminative ability of the questionnaire, we propose the use of different cutoff values for the GHQ-12 score for case identification depending on the age of each individual. PMID- 9406740 TI - Coping and cognition in schizophrenia and depression. AB - We examined the stable relations between coping style and cognitive function in schizophrenic and depressed patients and in patient and normal controls on two test occasions. The results show that a poor self-report of coping style is independent of psychiatric diagnosis, but there are associations with both subjective and objective cognitive malaise. Poor cognitive task performance is associated with a dependent coping style, perhaps pointing to a "giving-up" attitude. Subjective cognitive dysfunction and high levels of mental effort during task performance are associated with an avoidant coping style and with worrying, which suggest failing compensatory cognitive strategies as a causal mechanism of this coping dimension. PMID- 9406741 TI - Insight in seasonal affective disorder. AB - Lack of insight complicates the evaluation and treatment of patients with psychotic and affective disorders. No studies of insight in seasonal affective disorder (SAD) have been reported. Thirty patients with SAD diagnosed by the Structured Clinical Interview for DSM-III-R but no other axis I conditions were treated short-term with light-therapy. Insight was measured with the Scale to Assess Unawareness of Mental Disorder (SUMD) as modified by the authors to assess the self-report of insight into depressive symptoms. Increasing scores (1 to 5) indicated increasing unawareness of illness (i.e., less insight). SAD patients displayed a moderate amount of insight when depressed (mean SUMD score, 2.5). When recovered, they showed no significant change in insight into past depressive symptoms (mean SUMD score, 2.8). Greater insight into current depressive symptoms correlated with more depressive symptoms on the Hamilton Rating Scale for Depression score ([HRSD] r = .35, P < .05). In conclusion, SAD patients possess a moderate amount of insight into depressive symptoms that does not change after recovery, a result in agreement with studies of insight in psychosis and mania. Further, in SAD, increased severity of illness may be associated with increased insight into depressive symptoms, consistent with the hypothesis of depressive realism. PMID- 9406742 TI - An association between subcortical dementia and pernicious anemia--a psychiatric mask. AB - A case report from our psychiatric ward demonstrates the emergence of two different organic pathologies, a subcortical dementia and a pernicious anemia, preceded by a psychiatric presentation mimicking a dementia of depression. We emphasize that the psychiatric mask and the lack of other clinical and paraclinical cobalamin deficiency signs were the chief obstacles to early diagnosis. Only a complete and thorough assessment allowed the real diagnosis. We describe a subcortical dementia, and after a review of the literature, we support the causalty of B12 hypovitaminosis (pernicious anemia) in the genesis of this dementia, in the absence of other organic causes of mental syndromes and in the presence of a positive and well-maintained response to vitamin B12 administration. PMID- 9406743 TI - Clinical staging and operative reporting for multi-institutional trials in head and neck squamous cell carcinoma. AB - BACKGROUND: A Strategic Planning Conference (jointly supported by NCI and NIDCD) was convened to consider potential improvements in surgical patient data for multi-institutional trials. The thesis underlying this project is that inadequacies in staging, pretreatment patient stratification, and the details of surgical resection may have obscured the detection of treatment effect. The goals of this project were multiple: (1) to consider the utility of new clinical stratification variables, (2) to increase the precision of tumor staging, and (3) to improve operative reporting for multi-institutional trials in head and neck cancer. CONCLUSIONS: The conference attendees came to a number of important conclusions: (1) TNM status is inadequate for describing head and neck cancer in a multi-institutional trial setting. A detailed anatomic reporting scheme is proposed; (2) comorbidity measures should be included as patient descriptors, especially those that meet the criteria "definitely important and easy to obtain"; (3) surgical reporting in multi-institutional trials should use a format that is compatible with computer analysis and use the same items as the revised (anatomic) staging system; (4) the surgeon should be personally responsible for data coding and should interact directly with the pathologist in marking the surgical specimen; (5) pathologic reporting should use an anatomic template identical to the staging and operative reporting formats. PMID- 9406744 TI - Osseointegrated implants in the head and neck cancer patient. AB - BACKGROUND: Osseointegrated implants allow patients with oromandibular defects to obtain complete or partial dentition via implant-assisted or implant-borne prostheses. Implants restore masticatory and occlusal function, improving oral intake and articulation. However, use of implants in head and neck cancer patients has been discouraged due to lack of data supporting their utility in these patients. This study attempts to establish the validity of using osseointegrated implants for dental restoration in head and neck cancer patients. METHODS: Six patients who underwent resection/reconstruction for head and neck cancer received osseointegrated implants. Integration was assessed clinically, radiographically, and mechanically at 4-8 months; oral intake, mastication, and articulation were evaluated 6-12 months after receiving the dental prosthesis. RESULTS: Osseointegration occurred in 92% (24/26) of the implants: 100% (14/14) in neomandibles and 83% (10/12) in native mandibles. One patient had implants (2/5) that failed to integrate. The remaining patients' implants were immobile, free of infection, with no osteoradionecrosis. These patients tolerated a regular diet and experienced weight gain and improved articulation. CONCLUSIONS: The advent of osseointegrated implants and their compatibility with native and neomandible allows the restoration of functional dentition in patients undergoing ablative surgery for head and neck cancer. PMID- 9406745 TI - Evaluation of chemotherapy response in patients with advanced head and neck cancer using [F-18]fluorodeoxyglucose positron emission tomography. AB - BACKGROUND: [F-18]Fluorodeoxyglucose (FDG)-positron emission tomography (PET) can measure the metabolic activity of tissues; FDG-PET may be able to predict response to chemotherapy by identifying changes in tumor metabolism. Measurement of response to treatment may help improve survival in the management of advanced head and neck cancer. We evaluated this particular use of FDG-PET in patients participating in a neoadjuvant organ-preservation protocol using taxol and carboplatin and compared pathologic response after chemotherapy with changes in tumor metabolism measured by FDG-PET. METHODS: Serial FDG-PET studies (n = 56) were performed in patients (n = 28) with stage III/IV head and neck cancer participating in a neoadjuvant organ-preservation protocol. The FDG-PET studies were performed before and after chemotherapy. All patients had tissue biopsies before and after chemotherapy. Patients were classified as pathologic complete response (PCR) or residual disease (RD) based on tissue biopsies. Visual analysis of PET scans was performed to identify patients with complete response by PET, and these findings were compared with pathology results. Metabolic changes were also evaluated using standardized uptake ratios (SUR) of FDG. RESULTS: The sensitivity and specificity of PET for residual cancer after therapy was 90% (19/21) and 83% (5/6), respectively. Two patients had initially negative biopsies and positive PET studies for persistent disease. Pathology review and rebiospy led to confirmation of the PET results in these cases, giving a sensitivity of 90% for initial tissue biopsy. CONCLUSIONS: In this preliminary analysis, FDG-PET was accurate in classifying response to chemotherapy in most patients. Fluorodeoxyglucose-PET may identify residual viable tumor when it is otherwise undetectable. PMID- 9406746 TI - Baseline and post-treatment assessment of the general health status of head and neck cancer patients compared with United States population norms. AB - BACKGROUND: It is a common perception that the overall health of patients with head and neck cancer (HNC) is likely to be poor compared with the general population. This project was undertaken to investigate the pre- and post treatment, global health status of HNC patients in comparison with age-matched, U.S. population norms using a self-administered general health status survey. METHODS: Between July 1, 1993, and May 1, 1996, 180 patients underwent pretreatment and 6 month follow-up evaluation with the standard version of the Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36). The SF-36 scale scores, means, standard deviations, and 95% confidence intervals were calculated for each SF-36 scale as well as for physical-health-component summary scores (PCS) and mental-health-component summary scores (MCS). Comparisons of these scores were made to U.S. population normative data. Pretreatment and 6-month follow-up SF-36 scores were compared. RESULTS: In the 45-54-year age group, all 8 SF-36 scale scores, the PCS, and MCS scores were significantly worse for the HNC patients in comparison with age-matched norms (p < .05). In the 55-64-year age group, the HNC patients were worse in 5 of the 8 SF-36 scale scores and the MCS score in comparison with age-matched norms (p < .05). In the 65-74-year age group, the HNC patients scored significantly worse in the mental health scale. In the comparison of pretreatment and 6-month follow-up scores, the HNC patients had significant decreases in the physical functioning scale (p = .003) and the PCS score (p = .047). The HNC patients showed significant improvement in the mental health scale (p = .049) and improvement in the bodily-pain scale, which approached significance (p = .053) at 6-month follow-up. The HNC patients showed a marked decrease in general health status with increasing stage of HNC. CONCLUSIONS: This work provides objective support for the perception that many HNC patients are initially seen for treatment with baseline health status functioning significantly below their age-matched contemporaries in the general population. An educated evaluation of global health outcomes following treatment in the HNC patient population must begin with an accurate pretreatment assessment of these parameters. Self-reported health-status assessment (HSA) is a useful means of evaluating global health status in this patient population. PMID- 9406747 TI - Concomitant cisplatin/5-FU infusion and radiotherapy in advanced head and neck cancer: 8-year analysis of results. AB - BACKGROUND: The purpose of this study was to analyze long-term follow-up of a single institution's experience with a regimen of concomitant cisplatin/fluorouracil (5-FU) infusion and radiation given every other week. This analysis was stimulated by results of a randomized trial showing superiority for this regimen over induction cisplatin/5-FU chemotherapy followed by radiotherapy, especially in regional disease control. METHODS: All patients with stage III/IV disease who were referred by surgeons for nonoperative therapy and had a follow up of at least 2 years were included. Concomitant chemoradiotherapy was administered days 1-5 of a 2-week treatment cycle, for a total of 7 cycles, with cisplatin 60 mg/m2 day 1, 5-FU 800 mg/m2 given over 24 hours days 1-5, and radiation 2 Gy days 1-5. RESULTS: Seventy-eight patients with stage III (n = 16) or IV (n = 62) were treated and followed for a median of 8 years. Six patients died during treatment, of aspiration pneumonia, sudden death, gastrointestinal bleeding, and stroke. When assessed 6 weeks after the end of treatment, 45 patients (63%) had no clinical evidence of disease, whereas 27 (37%) still had some persistent abnormality. However, 17 of these "partial responders" have not recurred. In all, 24 patients (31%) have recurred or progressed, 13 at the primary site, 5 after 3 years. None of 16 stage III and 24 (39%) of 62 stage IV patients ever progressed. Tongue and glottic larynx did best, with only 1 of 22 patients ever failing (none locally). Supraglottic and oral cavity cancers other than tongue had the worst failure rates. Nineteen patients (24%) died of other causes (DOC), tumor-free. Patients who DOC correlated strongly with T stage (p < .002) but not with N stage or with AJC stage. The 5-year progression-free survival was 60% (confidence interval [CI] = 49% to 72%), and overall survival was 43% (CI = 33% to 56%). CONCLUSIONS: Disease control for this advanced head and neck cancer population was excellent. This regimen was especially effective in advanced tongue and glottic cancers and all stage III disease sites. Advanced supraglottic and hypopharynx cancers are problematic. These, and especially T4 lesions, are associated with high DOC rates, possibly in part related to swallowing malfunction. Nevertheless, the long-term survival without surgical intervention was high with this regimen. PMID- 9406748 TI - Has radiotherapy become too expensive to be considered a treatment option for early glottic cancer? AB - BACKGROUND: External beam radiotherapy and surgery produce equivalent long-term survival and tumor control in early glottic cancer. The expense and cost of radiotherapy have been challenged. METHODS: A retrospective review was performed for 57 patients undergoing radiotherapy for glottic cancer. End points included local tumor control, relapse-free survival, cause-specific survival, medical charges, and costs. The results were compared with those of 265 patients who underwent transoral endoscopic removal or an open laryngeal procedure at the same institution. RESULTS: The local control, larynx preservation, re-treatment, voice quality, relapse-free survival, and cancer death results and medical charges and costs are reported by treatment. CONCLUSIONS: Radiotherapy provides at least equivalent, if not superior, local tumor control, larynx preservation, voice quality, and survival, compared with the surgical options. Overall medical charges and costs for radiotherapy are similar to transoral endoscopic resection and less than partial vertical laryngectomy. PMID- 9406749 TI - Ectopic production of beta-HCG by a maxillary squamous cell carcinoma. AB - BACKGROUND: Paraneoplastic syndromes of the head and neck are rare. Hypercalcemia and leukocytosis have been described. The literature was reviewed, and a case of a squamous cell carcinoma of the maxilla producing beta human chorionic gonadotropin (beta-HCG) is presented. METHODS: A 47-year-old white man with a T4N1M0 squamous cell carcinoma of the left maxilla was treated with a maxillectomy and neck dissection for an N1 positive neck. After completing his planned radiotherapy, he developed distant metastases, which included an axillary node that stained positive for human beta-HCG. RESULTS: Retrospective review of the primary specimen showed beta-HCG positivity in an anaplastic component of the tumor along with vascular invasion. CONCLUSIONS: The first case in the literature of a paraneoplastic syndrome with beta-HCG production in association with squamous cell carcinoma of the maxilla is presented. This case history fits the aggressive nature of beta HCG producing tumors elsewhere in the body. PMID- 9406750 TI - Positron-emission tomography in parathyroid hyperplasia. PMID- 9406751 TI - Tracheal advancement flaps. PMID- 9406752 TI - Evidence for the calcium-dependent potentiation of M-current obtained by the ratiometric measurement of the fura-2 fluorescence in bullfrog sympathetic neurons. AB - Intracellular Ca2+ concentration ([Ca]i) was measured following the activation of an inward Ca2+ current and subsequent potentiation of an M-type K+ current (IM) in bullfrog sympathetic neurons. Fura-2 was used as an indicator for [Ca]i. The fluorescence ratio at 340 and 380 nm (F340/F380) was elevated from 0.36 to 1.22 when IM was potentiated by 68% following the Ca2+ current. Based on the in vivo calibration curve obtained from cells permeabilized with digitonin (20 microM), the F340/F380 value of 1.22 was equivalent to a [Ca]i of 0.97 microM. We therefore propose that a rise in [Ca]i into the micromolar range can lead to the potentiation of IM in amphibian autonomic neurons. PMID- 9406753 TI - Influence of respiratory afferents upon the proprioceptive reflex of skeletal muscles in healthy humans. AB - Relationships between respiratory afferents and the motor drive to skeletal muscles are well documented in animals, but human data are scarce. Tonic vibratory response (TVR) elicited by mechanical tendon vibrations were explored in an arm (extensor digitorum, ED) and a leg (vastus lateralis, VL) muscle, in healthy subjects. Tendon vibrations were delivered during unloaded breathing and after 10 breathing cycles while the subject breathed through an inspiratory or expiratory resistive load in order to activate respiratory afferents. Inspiratory loaded breathing significantly enhanced TVR in ED and VL muscles whereas the effects of expiratory loading depended on the vibrated muscle (increased TVR in ED; decreased TVR in VL). These results suggest that inspiratory muscle afferents activated during inspiratory loading facilitate the gamma motor drive to arm and leg muscles whereas the activation of pulmonary vagal afferents during expiratory loading can exert a facilitating or suppressive influence on the gamma motor drive, depending on the limb muscle group. PMID- 9406755 TI - Expansion of the neuropil of the mushroom bodies in male honey bees is coincident with initiation of flight. AB - The mushroom bodies (MB), the insect brain structures most often associated with learning, have previously been shown to exhibit structural plasticity during the adult behavioral development of female worker and queen honey bees. We now show that comparable morphological changes occur in the brains of male honey bees (drones). The volume of the MB in the brains of drones was estimated from tissue sections using the Cavalieri method. Brains were obtained from six groups of drones that differed in age and flight experience. Circulating levels of juvenile hormone (JH) in these drones were determined by radioimmunoassay (RIA). There was an expansion of the neuropil of the MB that was temporally associated with drone behavioral development, as in female queens and workers. The observed changes in drones were maintained in the presence of low levels of JH, also as in females. These results suggest that expansion of the neuropil of the MB in honey bees is associated with learning the location of the nest, because this learning is the most prominent aspect of behavioral development common to all members (workers, drones, queen) of the honey bee colony. PMID- 9406754 TI - Dopamine transporter mRNA is up-regulated in the substantia nigra and the ventral tegmental area of amphetamine-sensitized rats. AB - Converging evidence supports a significant role for dopamine (DA) in the development of behavioral sensitization and it has been suggested that changes in either DA transporter (DAT) or D2 autoreceptors contribute to the effects of stimulant treatment. To determine if alterations in DAT or D2 autoreceptor mRNA are long-lasting and parallel the time course of amphetamine (AMPH)-induced behavioral sensitization we performed the following experiment. Two groups of rats were used for mRNA analysis by in situ hybridization. They were given either single daily injections of saline or AMPH (2.5 mg/kg) for 5 days and sacrificed 7 days later. Two groups pretreated in a similar manner were used to test for behavioral sensitization. Pretreatment with AMPH which resulted in a sensitization response profile after AMPH challenge also produced a significant up-regulation of DAT mRNA in both the ventral tegmental area (VTA) (P = 0.01) and substantia nigra (SN) (P < 0.05) compared to the saline controls, whereas there were no significant group differences in D2 mRNA in either the SN or the VTA. The possible role of these changes in behavioral sensitization is discussed. PMID- 9406756 TI - Suppression of neuronal apoptosis by S-nitrosylation of caspases. AB - S-Nitrosylation (reaction of nitric oxide (NO) species with a critical cysteine sulfhydryl) can regulate the physiological activity of proteins, including enzymes, ion channels, G-proteins, and transcription factors. Caspases are a family of interleukin-1beta-converting enzyme-like proteases involved in the signaling pathway to apoptotic cell death, and each member of this enzyme family contains a critical cysteine residue in its active site. Here we show that S nitrosylation of caspases in human embryonic kidney (HEK)-293 cells and primary cerebrocortical neurons decreases enzyme activity and is associated with protection from apoptosis. PMID- 9406757 TI - Gait initiation and impairments of ground reaction forces as illustrated in old age by 'La marche a petits pas'. AB - Ground reaction forces during gait initiation and kinematics of the first step were recorded in 11 elderly patients with idiopathic 'marche a petits pas' and 18 age-matched normal adults. Smaller values of vertical forces, and impaired amplitudes and directions of anteroposterior forces might explain start difficulties of the patients. Higher vertical displacement of the foot, and laborious establishment of the rhythm in the patients could also be related to the perturbations observed in the reaction forces. These perturbations are likely to reflect impairments of muscular synergies of the lower limbs and the lack of limb co-ordination. PMID- 9406758 TI - Lack of effect of chronic clorgyline or selegiline on dopamine and serotonin transporters in rat caudate-putamen or nucleus accumbens septi. AB - Rats were injected intraperitoneally thrice weekly for 4 weeks with doses of selective inhibitors of monoamine oxidase type A (clorgyline, 1 mg/kg) or B ((-) selegiline, 10 mg/kg), or saline. Both treatments produced sustained elevations of concentrations of dopamine and serotonin, and decreased their deaminated metabolites in forebrain tissue. Nevertheless, no change in binding of [3H]GBR 12935 to the dopamine transporter or of [3H]paroxetine to the serotonin transporter in caudate-putamen or nucleus accumbens septi was found with quantitative autoradiography. These results support the impression that transporter proteins for these monoamines are not regulated by increased ligand abundance. PMID- 9406759 TI - Increase of cortical acetylcholine release after systemic administration of chlorophenylpiperazine in the rat: an in vivo microdialysis study. AB - The changes in acetylcholine (ACh) release from the cortex of freely moving rats after systemic administration of chlorophenylpiperazine (mCPP), a 5-HT2C agonist, were measured utilising microdialysis coupled to high performance liquid chromatography. mCPP administered intraperitoneally (i.p.) increased cortical ACh release, but failed to do so when applied locally in the cortex. The effect of i.p. administered mCPP on cortical ACh release was prevented by i.p. injection of mesulergine, a 5-HT2A/2C receptor antagonist, and isoteoline, a compound previously shown to antagonize behavioural effects of mCPP. An increase of cortical ACh release was also found after the local administration of mCPP in nucleus basalis magnocellularis (NBM). The results of the present work suggest that 5-HT2C receptors located in NBM are involved in the modulation of cortical ACh release in the rat. PMID- 9406761 TI - Increase of 5HT and VIP immunoreactivity within the hamster (Mesocricetus auratus) SCN during chronic MAOI treatment. AB - The effects of chronic treatment with the monoamine oxidase inhibitor (MAOI), clorgyline (CLG; 2 mg/kg per day) on serotonin (5HT) and vasoactive intestinal peptide (VIP) immunoreactivity (IR) within the hamster suprachiasmatic nucleus (SCN) were examined. Optical densities of 5HT IR and VIP IR were increased by MAOI treatment. VIP IR was increased in both the ventrolateral and dorsal regions of the SCN, suggesting that VIP content was increased within both perikarya and terminals of VIP neurons. The results suggest that previously described effects of MAOIs on the mammalian circadian system may be mediated in part, by their effects on serotonergic input to VIP neurons within the SCN. PMID- 9406760 TI - Evidence for a role of cholecystokinin as neurotransmitter in the guinea-pig enteric nervous system. AB - Intracellular recordings were made of neurons in the myenteric plexus of the guinea-pig distal ileum. Slow excitatory postsynaptic potentials (sEPSPs) were evoked by electrical stimulation of an interganglionic fibre tract. The effect of cholecystokinin (CCK) receptor antagonists on the sEPSPs was investigated in 11 neurons. Application of the CCK receptor antagonists L-364,718 and L-365,260 (each 250 nM) markedly attenuated the sEPSPs in five of 11 neurons. The amplitude of the sEPSP reduced from 15 +/- 3 to 7 +/- 2 mV and the change in membrane resistance during the sEPSP was reduced from 28 +/- 9 to 11 +/- 8 MS. In six of 11 neurons the CCK antagonists had no effect on the sEPSPs. The results provide evidence that neurally released CCK is involved in the mediation of sEPSPs in some enteric neurons. PMID- 9406762 TI - Electrophysiological responses of neurones in the rat nucleus tractus solitarius to oxytocin-releasing stimuli. AB - Neurones in the nucleus tractus solitarius (NTS) of the rat are, at least partially, responsible for mediating the excitatory effects of systemic cholecystokinin (CCK) injection and parturition on oxytocin release. Utilising in vivo electrophysiological recordings in urethane-anaesthetised rats we examined the effects of systemic CCK injection and vaginal distension (VD) on NTS neuronal activity. Approximately 25% of neurones were excited by CCK injection and a population of neurones (24%) responded specifically to VD, but only one neurone responded to both oxytocin-releasing stimuli. NTS neurones that projected to the hypothalamus also responded in a specific manner to either CCK or VD. It is known that the excitatory effects of CCK and VD on oxytocin release are mediated by separate peripheral pathways and the results from this study suggest that separation of these two inputs persists at the level of the NTS. PMID- 9406763 TI - NG108-15 cells induce the expression of muscular acetylcholinesterase when co cultured with myotubes. AB - Although muscular activity has been demonstrated to regulate the expression of acetylcholinesterase (AChE) in cultured myotubes, the exact role of the presynaptic terminus in regulating AChE expression at the neuromuscular junctions is not known. A chimeric co-culture of neuroblastoma x glioma hybrid NG108-15 cells with chick myotubes was established. By using chick-specific anti-AChE antibody, a protein of approximately 105 kDa in size corresponding to chick AChE catalytic subunit was revealed by Western blot analysis from the extracts of neuron-muscle co-cultures. In the co-cultures, NG108-15 cells induced the up regulation of muscle AChE expression by approximately 2.5-fold, while the control protein, chick muscle alpha-actinin at approximately 100 kDa, remained relatively unchanged. The NG108-15 cell-induced muscle AChE expression in the co-cultures was persistent when the muscular activity was blocked by alpha-bungarotoxin. In order to determine the AChE-inducing activity derived from NG108-15 cells, the cultured chick myotubes were treated with either conditioned medium of NG108-15 cells or its cell lysate. However, the muscle AChE, both in protein and activity levels, remained relatively unchanged. Our finding suggests that an AChE-inducing factor(s) is derived from the neuroblastoma cells in the co-cultures, but that may require the nerve-muscle contacts in culture. PMID- 9406765 TI - Analysis of the interrelations between a low-frequency and a high-frequency signal component in human neonatal EEG during quiet sleep. AB - It can be shown that dominant rhythmic signal components of neonatal EEG burst patterns (discontinuous EEG in quiet sleep) are characterised by a quadratic phase coupling (bispectral analysis). A so-called 'initial wave' (narrow band rhythm within a frequency range of 3-12 Hz) can be demonstrated within the first part of the burst pattern. The detection of this signal component and of the phase coupling is more successful in the frontal region. By means of amplitude demodulation of the 'initial wave' and a subsequent coherence analysis the phase coupling can be attributed to an amplitude modulation, i.e. the envelope curve of the 'initial wave' shows for a distinct period of time the same qualitative course as the signal trace of a 'lower' frequency component (0.75-3 Hz). The results were derived from six neonates (20 burst patterns for each neonate; 8 channel recordings). PMID- 9406764 TI - Evidence for the involvement of Na+-K+ pump and K+ conductance in the post tetanic hyperpolarization of the tetrodotoxin-resistant C-fibers in the isolated bullfrog sciatic nerve. AB - The post-tetanic hyperpolarizations (PTHPs) were recorded from the isolated bullfrog sciatic nerves using a modified vaseline gap method. Repetitive stimulation of tetrodotoxin-resistant (TTX-R) C-fibers produced frequency- and activity-dependent PTHPs. The ouabain solutions (0.1 mM), the solutions (0.1 mM ouabain + 2 mM tetraethylammonium (TEA)) and 5 mM TEA solutions reduced the PTHP amplitude to 56.2 +/- 10.7% (mean +/- SD, n = 9), to 37.0 +/- 12.3% (n = 8), to 76.0 +/- 10.9% (n = 5) of the control value, respectively. Each pair of the three solutions revealed a significant difference in their effects on the PTHP (P < 0.01). Furthermore, the Li-Ringer solutions and the solutions (Li-Ringer + 2 mM TEA) reduced the PTHP amplitude as well, thus resembling ouabain effects. These results strongly suggest that the repetitive stimulation of TTX-R C-fibers activates the membrane Na+-K+ pump resulting in a major part of the PTHP and induces TEA-sensitive K+ conductance composing a minor part of the PTHP, which might modulate repetitive firing of TTX-R C-fibers. PMID- 9406766 TI - Presidential address. The American Society of Parasitologists and Chandler's vision revisited. PMID- 9406767 TI - Acceptance of the 1997 Henry Baldwin Ward Medal. PMID- 9406768 TI - Some reflections on the teaching of parasitology. PMID- 9406769 TI - Biochemical properties of a neuraminidase of Trichomonas vaginalis. AB - Trichomonas vaginalis possesses a membrane-associated neuraminidase activity that is released into culture medium during its growth in vitro. The neuraminidase shows an optimum pH of 4.5 and a Km of 0.15 mM for 2'-(4-methylumbelliferyl) alpha-D-N-acetyl-neuraminic acid as a substrate. This enzyme releases mainly alpha-2,3-linked sialic acid because it is able to liberate sialic acid from sialyllactose (mainly alpha-2,3) but not from mucin (alpha-2,6) or fixed erythrocytes (mainly alpha-2,6). The neuraminidase activity is strongly inhibited by 2,3-dehydro-2-deoxy-N-acetyl neuraminic acid, whereas EGTA and Ca2+ do not affect the activity. Gel filtration-fast protein liquid chromatography of culture supernatant displays a single peak of neuraminidase activity with molecular weight 52,000. The levels of neuraminidase activity are variable in fresh and long-term grown isolates of T. vaginalis, regardless of time in culture. However, there are 2 kinds of isolates, 1 group with high neuraminidase activity and able to secrete the enzyme during growth and the other with low neuraminidase activity. The results suggest that T. vaginalis possesses a membrane-associated neuraminidase that is present to a variable degree in different isolates. PMID- 9406770 TI - Ultrastructure of spermatogenesis of Atriaster heterodus (Platyhelminthes, Monogenea, Polypisthocotylea). AB - The development of spermatozoa in the Polyopisthocotylea Atriaster heterodus was studied by transmission electron microscopy. Spermatogonia were undifferentiated, with irregular nuclei and little cytoplasm. Primary spermatocytes had sparse chromatin and typical synaptonemal complexes. The nuclear chromatin of secondary spermatocytes was in patches along the nuclear envelope and throughout the nucleoplasm. The complete process of fusion of the early spermatids to a common cytoplasmic mass forming a rosette was elucidated. Nuclei migrated to the center of the mass and changed from round to lamellar or tubular in shape. At the borders of the common cytoplasmic mass, the irregular zones of differentiation had microtubules, mitochondria, nuclei, centrioles, and intercentriolar bodies that give rise to 2 flagella. The spermatozoa presented a continuous row of cortical microtubules surrounding 2 parallels axonemes of the 9+1 type. PMID- 9406772 TI - Infrapopulation dynamics of Halipegus occidualis and Halipegus eccentricus (Digenea: Hemiuridae): temporal changes within individual hosts. AB - Individual infrapopulations of Halipegus occidualis and Halipegus eccentricus (Hemiuridae) in the buccal cavities of their amphibian host (Rana clamitans) were monitored for changes over time. Rates and patterns of parasite maturation, recruitment, and loss were estimated, and the probabilities of infection for definitive hosts in this system were calculated. Although variable, the time required for maturation of immature worms (once in the buccal cavity) was as little as 1 wk. Relatively rapid losses of worms from individual infrapopulations were observed, although this did not appear to inhibit additional recruitment of these trematodes. Also, it was clear that these species overwintered within their amphibian hosts. There was no statistical difference among the 3 sampling years in the probability of acquiring an initial or a subsequent infection with either parasite. However, we suggest that frogs with an existing infrapopulation may have a greater probability of adding worms if those hosts are within certain foci of infection within the pond. The probability of frogs becoming infected in this system increased in late spring and peaked in midsummer. By examining the temporal changes within individual infrapopulations, it is clear that dynamic changes in infrapopulation size and composition may occur, suggesting that rates of trematode transmission may be greater than previously estimated. PMID- 9406771 TI - Development of Eimeria ninakohlyakimovae Yakimoff & Rastegaieff, 1930 emend. Levine, 1961 in experimentally infected goats (Capra hircus). AB - The endogenous development and prepatent and patent periods of Eimeria ninakohlyakimovae were studied in 43 1-3-wk-old coccidia-free kids inoculated with 5.0 x 10(4), 1.5 x 10(5), 2.0 x 10(5), or 9.0 x 10(5) sporulated oocysts/kg. Twenty-five kids were killed at 24- or 48-hr intervals, 2-18 days after inoculation (DAI). Two generations of meronts, gamonts, gametes, and oocysts were found in sections stained with hematoxylin and eosin and examined using under light microscopy. The first generation of meronts developed in the endothelium of the lacteals, in the lamina propria, and in the lymphatic vessels of the ileum submucosa. Mature, first-generation meronts, 165.5 x 123.6 microm, were first found 10 DAI. Second-generation merogony developed in the crypt epithelial cells of the cecum and colon; mature meronts, 16.8 x 11.6 microm, were first seen 12 DAI. Gametogenesis occurred in the cecum and colon epithelium; mature microgamonts (16.1 x 13.0 microm), microgametes, macrogametes (14.7 x 12.5 microm), and oocysts (18.3 x 13.3 microm) were seen at 13 DAI. The course of the infection was followed in 18 kids examined every day until 24 DAI. The prepatent period was 14.7 (13-17) days and the patent period 6.8 (4-10) days. The sporulation time at 30 C, with constant aeration, was 2-3 days. PMID- 9406774 TI - PCR and DNA hybridization indicate the absence of animal filariae from vectors of Onchocerca volvulus in Uganda. AB - In order to identify Onchocerca volvulus larvae from vectors, DNA of filaria larvae from dissected blackflies was isolated, and a 150-bp long tandemly repeated DNA sequence (0-150), which occurs in many Onchocerca species, was amplified using polymerase chain reaction (PCR). Subsequently, the PCR product was blotted onto a nylon membrane and hybridized with DNA probes specific for O. volvulus or Onchocerca ochengi. Filaria larvae from 395 infected Simulium neavei were examined and 259 samples produced detectable PCR products. Among these samples, 239 (92%) reacted with an O. volvulus-specific oligonucleotide. A sample of 69 PCR products was tested using an O. ochengi DNA probe, but all failed to hybridize. Filaria larvae from 64 infected Simulium damnosum, presumably of the cytotypes "Nyamagasani" and "Nkusi" were studied and 0-150 was amplified from 38 samples. From these samples, 35 (92%) hybridized specifically with an O. volvulus probe but none with the O. ochengi-specific DNA sequence. Nonamplified samples were obtained mainly from blackflies that contained only 1 or 2 filaria larvae, and therefore, an insufficient DNA extraction was assumed. It can be concluded that few, if any, filaria species of animal origin were transmitted by S. neavei and S. damnosum s.l. in Kabarole and Kasese districts in Uganda. PMID- 9406773 TI - Seasonal occurrence of Gyrodactylus derjavini (Monogenea) on brown trout, Salmo trutta, and Atlantic salmon, S. salar, in the Sandvikselva river, Norway. AB - Seasonal variation in prevalence and intensity of the ectoparasite Gyrodactylus derjavini on brown trout parr, Salmo trutta, and Atlantic salmon parr, S. salar, and occurrence of G. derjavini on ascending sea trout, S. trutta m. trutta, in the Sandvikselva river, southeastern Norway, was recorded. In general, both prevalence and intensity increased and decreased correspondingly with the rise and fall in water temperature. However, both prevalence and intensity decreased in warm periods when reproduction and transmission should be high, possibly from host-induced parasite mortality. Sea trout became infected with G. derjavini soon after they entered the river; prevalence tended to increase as the sea trout migrated upstream to the spawning grounds. It is hypothesized that transmission of G. derjavini to sea trout occurred via the river substratum. PMID- 9406775 TI - Evolution of the schistosomes (Digenea: Schistosomatoidea): the origin of dioecy and colonization of the venous system. AB - Trematodes of the family Schistosomatidae are considered venous system specialists whose sister group is the vascular system generalists (Spirorchidae) of turtles. Colonization of homeotherms by vascular trematodes required precision egg laying near the conduit for egg passage to the external environment and avoidance of pathogenesis that might result in the premature death of the host. Evolution of dioecy from the hermaphroditic condition may have proceeded through androdioecy in which hermaphrodites were specialized for precision egg placement in the vascular system and larger adults became functional males. The evolution of nuclear genes suppressing female function along with cytoplasmic genes suppressing male function could then have resulted in the origin of dioecious, dimorphic populations. Schistosomes compensated for the reduction in potential reproductive partners by (1) increased overdispersion in the vertebrate host, (2) reduced egg hatching time in the external environment, (3) formation of permanent pairs mimicking the hermaphroditic condition, (4) increased longevity in the definitive host, and (5) increased fecundity. Colonization of the venous system was necessitated by (1) evolutionary radiation into terrestrial vertebrates and (2) the increased immunopathology associated with the high, constant body temperature of homeothermic vertebrates. The immune response to spirorchid and schistosome eggs appears to be qualitatively similar in their respective hosts. The arterial dwelling spirorchids release eggs in the direction of blood flow, resulting in a wide dissemination of eggs within the host. The lower body temperature of poikilotherms accompanied by the seasonal nature of the immune response in these hosts would result in a quantitatively reduced pathogenesis. Hosts that did succumb to the infection would most likely die in water, where eggs could be released by predation, scavengers, or decomposition and develop successfully. Colonization of the venous system by schistosomes would require precision egg placement because eggs are released against blood flow. Eggs are sequestered within the portal system of homeotherms, thus restricting egg dispersal and resulting pathogenesis to less sensitive organs. A significant number of eggs may escape into the external environment before a heavily infected host is incapacitated by, or dies from, the infection. PMID- 9406776 TI - Limited range of genetic variation in Echinococcus multilocularis. AB - DNA sequencing of 1.3 kb of rDNA containing both internal transcribed spacers (ITS1, ITS2) and adjoining rRNA coding regions in each of 11 Echinococcus multilocularis isolates from Germany, Japan, and Alaska resulted in identical nucleotide sequences except for a single polymorphic locus 54 bp upstream of the 3' end of the 18S coding region, separating Eurasian isolates from an Alaskan isolate. The same base substitution was found in each of 2 additional isolates from Alaska. The distribution of the resulting genotypes with regard to their origin is highly significant (>99.9%) and corresponds to the traditional subspecies Echinococcus multilocularis multilocularis and Echinococcus multilocularis sibiricensis. PMID- 9406777 TI - Quantification of cytokine gene expression in lamina propria lymphocytes of cattle following infection with Ostertagia ostertagi. AB - Changes in cell surface markers and cytokine transcription were analyzed in lamina propria lymphocytes from control animals (noninfected calves) and calves after a single high but nonprotective primary infection with Ostertagia ostertagi. Flow cytometry of cells recovered from the lamina propria showed an increase in the percentages of IgM+, WC1+, and IL-2R+-bearing cells 10 days after infection; however, 2 mo after infection, cell staining was comparable to preinfection levels. Transcription levels of interleukins IL-2, IL-4, IL-10, and interferon (IFN)-gamma mRNA were measured using a competitive reverse transcription-polymerase chain reaction. Results indicated elevated levels of IL 4 and IFN-gamma in the infected animals at 10 days and at 60 days after infection. Transcription of IL-10 also increased; however, this change was not observed until 60 days postinfection. PMID- 9406778 TI - Prevalence of antibodies to Neospora caninum in different canid populations. AB - A total of 1,554 dogs from 5 countries on 3 continents were tested for antibodies to Neospora caninum using an indirect fluorescent antibody test. In Australia, overall, 42/451 (9%, 95% confidence interval [CI] 6-12%) dogs were seropositive (Melbourne 11/207 [5%, 95% CI 2-9%]; Sydney 18/150 [12%, 95% CI 7-18%]; Perth 13/94 [14%, 95% CI 8-22%]). Antibodies to N. caninum were also detected in dogs in South America (Uruguay [20%, 95% CI 16-24%, n = 414]) and sub-Saharan Africa (Tanzania [22%, 95% CI 12-36%, n = 49]). In contrast, only 1 of 500 dogs tested from the Falkland Islands and none of 140 dogs from Kenya was seropositive. Of wild canids, 1/54 (2%, 95% CI 0-10%) British foxes and 15/169 (9%, 95% CI 5-14%) Australian dingoes had antibodies to N. caninum. PMID- 9406779 TI - Trypanosoma cruzi: effect of immunization on the risk of vector-delivered infection in guinea pigs. AB - The protective effect of experimental immunization was studied in guinea pigs exposed to vectorial infection by Trypanosoma cruzi. Immunized animals received an inoculum of live-attenuated T. cruzi epimastigotes into a granuloma previously induced by Freund's complete adjuvant in the hind footpad. Seven days later, a delayed-type hypersensitivity reaction was triggered by reinjection of the parasites in the front footpad. The animals were then placed in Triatoma infestans-colonized corrals and exposed to vectorial T. cruzi transmission of the parasite for up to 200 days. The effectiveness of this immunizing protocol was controlled in terms of the number of bites necessary for infection (NBNI) in immunized as compared with control animals. Periodic entomological census allowed for the determination of vector biting and infection rates and the calculation of NBNI. Although this measurement was quite variable between yards, an overall average of 4,973 bites was enough to infect a control guinea pig in 4 separate experiments. The corresponding figure for the experimental group was 21,307 bites, implying that immunized animals could resist a 4.28-fold increase (range: 1.99-8.32) in the number of vector bites before becoming infected. PMID- 9406780 TI - Antibody responses of cows during an outbreak of neosporosis evaluated by indirect fluorescent antibody test and different enzyme-linked immunosorbent assays. AB - Serum samples from 70 (33 aborting and 37 non-aborting) dairy cows from a herd in California were analyzed for Neospora caninum antibodies in different laboratories by various serologic assays including enzyme-linked immunosorbent assay (ELISA) with recombinant antigens (Nc4.1 and Nc14.1), kinetic ELISA, whole tachyzoite lysate ELISA, immunostimulating complex (iscom) ELISA, antigen capture competitive inhibition ELISA, and by the indirect fluorescent antibody test. Eighteen percent of pregnant cows in this herd had aborted within 2 mo of the index case. All 70 cows had antibodies to N. caninum by at least 1 of the tests. Antibody levels to N. caninum in aborting cows as a group were higher than in nonaborting cows. However, it was concluded that no serological test could be used to establish definitively that N. caninum caused the abortion in an individual cow. PMID- 9406781 TI - Kinetics in parasite abundance in susceptible and resistant mice infected with an avirulent strain of Toxoplasma gondii by using quantitative competitive PCR. AB - The kinetics of changes in Toxoplasma gondii abundance were evaluated with a quantitative competitive (QC)-polymerase chain reaction (PCR) assay at various sites in both C57BL/6 and BALB/c mice. Higher mortality was apparent in C57BL/6 mice than in BALB/c mice when infected with a high dose of cysts. There were significant differences in cyst number when infected with a low dose of cysts, although there was no significant difference in mortality between the 2 mouse strains. One day after infection with a low dose of an avirulent Fukaya strain, T. gondii was detected in peripheral blood, mesenteric lymph nodes, spleen, lungs, and brain. Two weeks after infection, the number of T. gondii in the brain greatly increased in C57BL/6 mice but not in BALB/c mice. Thus, it would appear that the first to second week after infection is a critical period in determining T. gondii abundance. QC PCR allows the detection of low numbers of T. gondii at an early stage of infection in the murine model. This is useful for the early diagnosis of toxoplasmosis and to understand reactivation of toxoplasmosis. PMID- 9406782 TI - Detection of Paragonimus heterotremus in experimentally infected cat feces by antigen capture-ELISA and by DNA hybridization. AB - An antigen capture enzyme-linked immunosorbent assay (antigen capture-ELISA) and DNA hybridization technique were developed and evaluated for their application in the detection of Paragonimus heterotremus infection in experimentally infected cats. An IgG fraction prepared from serum of a rabbit immunized with P. heterotremus excretory-secretory (ES) products was used as the capture antibody. An IgG1 monoclonal antibody specific to the 22- and 31.5-kDa ES products of P. heterotremus was used as the antigen probe. As little as 0.24 ng of the ES products could be detected by this technique. A specific P. heterotremus DNA probe derived from the P. heterotremus genomic DNA library containing 1,500 base pairs was used in a dot-blot hybridization assay for the detection of parasite DNA. The radioactively labeled probe could detect DNA released from as few as 2 P. heterotremus eggs. Both ELISA and DNA hybridization were found to have 100% specificity, with sensitivities of 73.7% and 100%, respectively. PMID- 9406783 TI - Schistosoma mansoni cercariae: stimulation of acetabular gland secretion is adapted to the chemical composition of mammalian skin. AB - The chemical signals of mammalian skin that stimulate the secretion of acetabular gland contents of Schistosoma mansoni cercariae were determined by exposing cercariae to fractions of human and pig skin surface obtained by thin-layer chromatography. Postacetabular gland secretion was stimulated by hydrophilic skin extracts but was often combined with a secretion of preacetabular glands. Secretion of preacetabular glands, which contain enzymes for skin lysis, could be selectively stimulated with skin surface lipids. Two different mechanisms of lipid-stimulated preacetabular gland release could be distinguished. First, secretion in combination with penetration behavior and probably tegument transformation was stimulated by the fraction of free fatty acids. Second, secretion independent of penetration behavior and tegument transformation was exclusively stimulated by glucosylceramides and phospholipids, probably phosphatidylcholines. The secretion mechanisms seem to allow a continuous lysis of epidermal macromolecules during the skin passage of the cercariae. Free fatty acids occur in the uppermost skin layers and may stimulate the combination of the first response; phospholipids and glucosylceramides are restricted to deeper epidermal layers and may stimulate the enzyme secretion there. An active preacetabular gland release was also stimulated by toxic chemicals, which could suggest an emergency penetration program for impaired cercariae. PMID- 9406784 TI - SSU rDNA characterization of lymnaeid snails transmitting human fascioliasis in South and Central America. AB - The small subunit (18S) rRNA gene sequences of the lymnaeid morphs I and II (Mollusca: Gastropoda: Basommatophora: Lymnaeidae) transmitting human fascioliasis in the high endemic zone of the northern Bolivian Altiplano and of Lymnaea cubensis from Mexico and Guadeloupe island (Caribbean) have been obtained by direct polymerase chain reaction PCR cycle sequencing and silver staining methods and compared to that of the 6 most common European Lymnaeidae species. Results allow us to establish definitively the distinction between the lymnaeids from the northern Bolivian Altiplano and L. cubensis. Lymnaea cubensis is a valid species distributed in North and Central America but absent in the northern Bolivian Altiplano. Lymnaeid morphs I and II from the northern Bolivian Altiplano both present the same 18S rDNA sequence, which is moreover identical to that of the European species Lymnaea truncatula. Significant nucleotide substitutions in helix E10-1 of the variable region V2 of the secondary structure suggest the need for distinguishing L. cubensis in the subgenus Lymnaea (Bakerilymnaea) with L. (B.) cubensis as type species. The subgenus Lymnaea (Fossaria) is retained, with L. (F.) truncatula as type species. The larger number of nucleotides in the 18S rDNA sequence of L. cubensis (1,860 bp) with regard to the other Lymnaea species (1,843-1,852 bp) is tentatively related to the more ancient paleogeographic origin of L. cubensis. The grouping of L. cubensis with L. truncatula and the relationship of Lymnaea auricularia with Lymnaea peregra in the phylogenetic trees obtained show an evolutionary parallelism with the digenean parasite species they transmit, namely Fasciola hepatica and Fasciola gigantica, respectively. PMID- 9406786 TI - Infection of Anopheles freeborni mosquitoes on New World monkeys infected with the Uganda I/CDC strain of Plasmodium malariae. AB - Anopheles freeborni mosquitoes fed during 85 primary and 26 recrudescent infections of the Uganda I/CDC strain of Plasmodium malariae in Saimiri and Aotus monkeys were examined for the presence of oocysts. Of these, 42 primary and 14 recrudescent infections were infective. Mosquitoes were more frequently infected when fed upon A. lemurinus griseimembra animals. A retrospective examination indicated the greatest mosquito infectivity occurred before the maximum parasite count. Mosquito infection was highest 4, 5, and 6 days after the parasite count exceeded 1,000/microl. Overall, 98 of 304 positive lots (32.2%) had > or = 50% of the individual mosquitoes infected. In addition, lots of An. freeborni were fed through membranes on the blood of 34 monkeys. During the days following the parasite count reaching > or = 1,000/microl, feedings on the animals resulted in lower levels of infection than membrane feeding, thus extending the period of mosquito infection. PMID- 9406785 TI - Acid phosphatase activity in Perkinsus marinus, the protistan parasite of the American oyster, Crassostrea virginica. AB - The effect of temperature (4, 12, 20, and 28 C) and osmolality (400, 570, and 840 mOsm/kg) on extracellular acid phosphatase (AP) secretion in vitro, and ultrastructural localization of AP activity in the parasite were investigated. The extracellular AP secretion by Perkinsus marinus was cell density dependent (P < 0.001). Increasing culture temperatures resulted in increased P. marinus proliferation concomitant with AP secretion (P < 0.0001). AP secretion was similar in P. marinus cultured at 400 and 570 mOsm/kg media, but higher than P. marinus cultured at 870 mOsm/kg media. Results of the ultrastructural study revealed that intense AP activity was in the nucleus of the parasite. Based on its distribution in the nucleus, AP may be playing a role in events leading to cell cycle regulation. PMID- 9406787 TI - Susceptibility of Peromyscus leucopus and Mus musculus to infection with Baylisascaris procyonis. AB - In this study, we compared the susceptibility of Peromyscus leucopus (white footed mouse), a common natural intermediate host, and Mus musculus, a commonly used experimental model, to infection with larvae of the raccoon ascarid, Baylisascaris procyonis. Three groups of 10 mice of each species were given 50, 250, or 500 infective B. procyonis eggs by gavage. The mice were observed daily for clinical signs of central nervous system (CNS) disease and at necropsy the distribution of larvae in 10 body regions and organs was determined and compared. Clinical CNS disease developed in 57% of P. leucopus and 93% of M. musculus. The average clinical incubation period was significantly longer in P. leucopus (20.6 days postinfection [PI]) than in M. musculus (10.7 days PI), and clinical disease progressed slower in P. leucopus. Significantly fewer larvae were recovered from P. leucopus than from M. musculus. Most larvae were recovered from the anterior carcass and viscera of P. leucopus and from the carcass, head, and brain of M. musculus. CNS invasion was dose dependent in M. musculus but not in P. leucopus. Few or no grossly visible larval granulomas were present in P. leucopus but were abundant in M. musculus. We concluded that P. leucopus was less susceptible than M. musculus to B. procyonis infection, based on a decreased intensity of infection, longer clinical incubation period or lack of clinical disease, slower progression of disease, different larval distribution, and lower tissue reactivity to larvae. PMID- 9406790 TI - Himasthla limnodromi n. sp. (Digenea: Echinostomatidae) from the short-billed dowitcher, Limnodromus griseus (Aves: Scolopacidae). AB - Himasthla limnodromi n. sp. is described from short-billed dowitchers, Limnodromus griseus, from the Araya Peninsula, Venezuela, and Delaware Bay, U.S.A. Himasthla limnodromi was not found in dowitchers on the breeding grounds or on the fall staging grounds in the Bay of Fundy but reappeared in dowitchers on the wintering grounds in the fall. This suggests that H. limnodromi is acquired by the birds on arrival on the wintering grounds and then gradually disappears during the birds' northward migration in the spring. The new species has a reniform collar armed with 31 spines, with 23 in a single uninterrupted row, and 4 corner spines in overlapping pairs at each end. The cirrus sac is up to 10 times longer than the length of the acetabulum and contains a long, smooth cirrus. The vitellaria always commence posterior to the posterior end of the cirrus sac in mature specimens. The testes are found in the posterior eighth of the long, filamentous body. Himasthla limnodromi n. sp. most closely resembles Himasthla alincia, but H. limnodromi is larger in size and has an unspined cirrus and smaller eggs. PMID- 9406789 TI - Effect of gamma radiation on Brugia L3 development in vivo and the kinetics of granulomatous inflammation induced by these parasites. AB - Previous studies have shown that the downregulation of parasite-specific cellular immune response in Brugia-infected jirds requires viable worms but is not dependent on microfilariae (MF) for either induction or maintenance of this phenomenon. To clarify further which life cycle stages induce filarial hyporesponsiveness, jirds were infected intraperitoneally with third stage larvae (L3) exposed to 0, 15, 25, 35, 45, or 90 krad of gamma radiation to differentially alter L3 development. Necropsies were performed at 7, 14, 28, and 118 days postinoculation (DPI). The degree of parasite development, intraperitoneal inflammation, and pulmonary granulomatous inflammation (PGRN) to parasite antigen-coated beads embolized in the lungs were monitored at the time of necropsy. Parasite survival and worm lengths were inversely related to the irradiation dose. Gamma radiation at 35, 45, or 90 krad prevented larval molt to the adult stage. Some parasites irradiated with 15 or 25 krad developed beyond fourth stage larvae (L4) to infertile adult females. The PGRN peaked at 14 DPI in all infected groups. Downregulation of the PGRN occurred after 14 DPI in groups that received nonirradiated L3 or L3 irradiated with 15 krad. No significant decrease of the PGRN occurred in groups that received parasites irradiated with more than 15 krad. Significant peritoneal inflammation as indicated by an increase in macrophages occurred only in jirds that received nonirradiated L3. These data demonstrate the importance of the adult stages in inducing downmodulation in the absence of MF and suggest that the L4 may also play a role in the induction of this phenomenon. An alternate conclusion is that parasite burden and not developmental stage is important in the induction of this hyporesponsive state. PMID- 9406788 TI - Affinity isolation and characterization of the cathepsin B-like proteinase Sj31 from Schistosoma japonicum. AB - The major cathepsin B-like proteinase of adult Schistosoma japonicum has been isolated for the first time. Affinity chromatography with the mammalian cathepsin B inhibitor glycyl-phenylalanyl-glycine-semicarbazone purified a protein that was identified by N-terminal sequencing as Sj31. Sensitivity of Sj31 to PNGase F demonstrated the presence of asparagine-linked N-glycan. Marked resistance to the action of Endo-beta-glycosidase H indicated that most of the N-glycan chains are of the complex type. Binding of horseradish peroxidase-conjugated lectins demonstrated the presence of N-mannose, N-acetylglucosamine, and N acetyllactosamine type 2 in the N-glycan. Fucose was not detected, and the presence of sialic acid remained questionable. Sj31 degraded the fluorogenic substrates Z-Phe-Arg-NMec and Z-Arg-Arg-NMec with an optimum between pH 5.0 and 6.0. The specific activity was 18-21-fold higher with the Phe-Arg substrate compared with the Arg-Arg substrate, whereas this value was 4-6-fold for bovine spleen cathepsin B, thus suggesting differences in the S2 subsite between parasite and host proteinases. Quantitative purification of Sj31 led to the conclusion that cathepsin B-like activity predominates over cathepsin L-like activity in S. japonicum. Because Sj31 degraded hemoglobin in vitro and was localized in the parasite gut, the proteinase may degrade ingested proteins in vivo. PMID- 9406794 TI - Crepidostomum percopsisi n. sp. (Digenea: Allocreadiidae) from the trout perch (Percopsis omiscomaycus) of Dauphin Lake, Canada. AB - Crepidostomum percopsisi n. sp. is described from the small intestine of the trout perch (Percopsis omiscomaycus) in Dauphin Lake, Manitoba. It is morphologically similar to Crepidostomum isostomum, which has been reported from the trout perch and several other species of fish. It differs from C. isostomum based on the vitellaria confined to the hindbody of the worm, size and shape of the cirrus, size of the testes, and its greater body length. A comparison of our specimens with those illustrated and identified as C. isostomum from trout perch indicates that such specimens are identical to larger specimens of C. percopsisi recovered by us from trout perch in May. To date, C. percopsisi has only been reported from the trout perch of Dauphin Lake, Lake Winnipeg, and Oneida Lake, which suggests host specificity. PMID- 9406793 TI - Parvicapsula minibicornis n. sp. (Myxozoa, Myxosporea) from the kidney of sockeye salmon (Oncorhynchus nerka) from British Columbia, Canada. AB - A new species of Myxosporea Parvicapsula minibicornis is described from the kidney of adult sockeye salmon (Oncorhynchus nerka) that had recently returned from the Pacific Ocean to Weaver Creek, a tributary of the Fraser River, British Columbia, Canada. Spores are ovoid, symmetrical, with 2 pyriform polar capsules at the anterior pole. The posterior pole has 2 small, pointed projections. Mean spore dimensions are length 11.0 microm, thickness (perpendicular to suture plane) 6.8 microm, and width (in sutural plane) 7.5 microm. This myxozoan is compared to other described Parvicapsula species and a Parvicapsula sp. from netpen-reared coho salmon (Oncorhynchus kisutch). PMID- 9406791 TI - Phylogeny of the orders of the Eucestoda (Cercomeromorphae) based on comparative morphology: historical perspectives and a new working hypothesis. AB - The phylogeny of the Eucestoda was evaluated based on a suite of 49 binary and multistate characters derived from comparative morphological and ontogenetic studies; attributes of adult and larval tapeworms were considered. A single most parsimonious tree (MPT) (consistency index = 0.872; retention index = 0.838; and homoplasy index = 0.527) was fully resolved and is specified by the following: (Gyrocotylidea, (Amphilinidea, ((Spathebothriidea, (Pseudophyllidea, ((Diphyllidea, (Trypanorhyncha, (Tetraphyllidea, (Lecanicephalidea, ((Nippotaeniidea, (Tetrabothriidea, Cyclophyllidea)), Proteocephalidea))))), Haplobothriidea))), Caryophyllidea))). Monophyly for the Eucestoda was firmly corroborated. Trees derived from the primary and bootstrap analyses were congruent, but low values, particularly for relationships among the tetrafossate tapeworms, indicated additional examination is warranted. The MPT was found to be the most efficient hypothesis for describing character evolution and in specifying relationships among the orders when compared to those concepts that had been developed for the tapeworms over the past century. Areas of congruence were shared among the current hypothesis and one or more of the prior hypotheses. Major conclusions include: (1) Caryophyllidea are basal and monozooy is ancestral; (2) difossate forms are primitive, and the Pseudophyllidea are the sister group of the strongly polyzoic tapeworms; (3) Nippotaeniidea are highly derived; (4) the higher tapeworms (Tetraphyllidea, Lecanicephalidea, Proteocephalidea, Nippotaeniidea, Tetrabothriidea, and Cyclophyllidea) are closely related or potentially coordinate groups: (5) Tetrabothriidea and the Cyclophyllidea are sister groups; and (6) Tetraphyllidea is paraphyletic, with the Onchobothriidae basal to the Phyllobothriidae. Character support for placement of the Tetrabothriidea continues to be contradictory, and this order may represent a key to understanding the phylogeny of the higher cestodes. The current study constitutes a complete historical review and poses a new and robust hypothesis for the phylogeny of the Eucestoda. PMID- 9406792 TI - Redescription of Sarcocystis levinei Dissanaike and Kan, 1978 (Protozoa: Sarcocystidae) of the water buffalo (Bubalus bubalis). AB - Sarcocystis levinei Dissanaike and Kan, 1978, is redescribed because the original description was a mixture of 2 species, the amended S. levinei and the newly recognized S. buffalonis Huong, Dubey, Nikkila, and Uggla, 1997. In histological sections, S. levinei sarcocysts are microscopic, up to 640 microm long and up to 95 microm wide. Ultrastructurally, the cyst wall is thin (< 1.0 microm thick) with a minute undulating surface and smooth, hairlike villar protrusions arising at irregular distances from the cyst wall. The villar protrusions have a dome shaped base (approximately 0.5 microm thick), a fingerlike middle part, and a tapering distal end (<0.1 microm thick). The morphological features of the sarcocyst resemble those of S. cruzi (Hasselman, 1926) Wenyon, 1926, of cattle. Sarcocystis levinei sarcocysts were found in striated muscles including heart, esophagus, tongue, and skeletal muscle. The buffalo myocardium is parasitized exclusively by S. levinei, whereas >1 Sarcocystis species may occur concurrently in other muscular tissues of water buffaloes. Two dogs, but not 2 cats, fed water buffalo hearts infected with S. levinei sarcocysts shed sporocysts measuring 9.5 10.5 x 14.0-16.5 microm starting from days 16 and 18, respectively. PMID- 9406795 TI - Redescription of Camallanus cotti Fujita, 1927 (Nematoda: Camallanidae) from Hawai'i. AB - The freshwater fish parasite Camallanus cotti Fujita, 1927 (Nematoda: Camallanidae) is redescribed from the guppy Poecilia reticulata (Poeciliidae). We confirm previous reports of its occurrence in other introduced poeciliids in Hawai'i, in 4 species of native Hawaiian gobioid stream fishes, and in an elasmobranch, an aquarium-reared stingray Potamotrygon sp. (Dasyatididae) from Hawai'i. Because the source localities of introduced freshwater fish parasites may be far removed geographically from communities that they have invaded, especially oceanic archipelagos (such as Hawai'i), we believe that research on these exotic parasites must be based upon critical taxonomic evaluations. Our redescription of C. cotti serves as a foundation upon which ecological studies of this parasite, performed in conjunction with conservation efforts for native Hawai'ian stream fishes, will be based. PMID- 9406796 TI - A new Hepatozoon species from dogs: description of the causative agent of canine hepatozoonosis in North America. AB - A new species of Adeleina, Hepatozoon americanum, is described from the skeletal muscle, cardiac muscle, visceral organs, and blood of dogs (Canis familiaris) in the Southern United States. The organism was previously identified as Hepatozoon canis (James, 1905) Wenyon, 1926; however, differences in clinical signs, histopathological and serological findings, gamont size, and ultrastructure define the new species of Hepatozoon. Attempts to transmit the protozoan from infected dogs to nymphal Rhipicephalus sanguineus ticks, the definitive host of H. canis, were not successful. PMID- 9406797 TI - The effect of Gyrodactylus salaris (Monogenea) on the epidermis of Atlantic salmon, Salmo salar, parr in the river Batnfjordselva, Norway. AB - Skin morphology of Atlantic salmon, Salmo salar, parr infected with Gyrodactylus salaris was examined for 3 yr in the river Batnfjordselva in Norway and compared to that of uninfected salmon parr from a neighboring river. The epidermis of the infected population had more cell layers and was thicker than the epidermis of parr from the uninfected population. The number of mucous cells did not differ, and no seasonal changes in morphology of the epidermis were detected in either rivers. Intensity of G. salaris did not correlate to epidermal thickness, epidermal cell layers, or mucous cell concentration. PMID- 9406798 TI - Adaptation of a strain of Plasmodium falciparum from a Montagnard refugee to Aotus monkeys. AB - A strain of Plasmodium falciparum from a Montagnard refugee was shown to produce large numbers of gametocytes in culture. Attempts were made to establish this strain in Aotus monkeys via trophozoite and sporozoite inoculation. The Montagnard S-1 strain was readily adapted to A. l. griseimembra monkeys via trophozoite inoculation. Other species of Aotus failed to support the development of high density parasitemia. None of 12 attempts to transmit the infection via sporozoites from Anopheles freeborni or An. dirus mosquitoes was successful; however, developing exoerythrocytic stages were demonstrated in hepatocytes of an A. lemurinus griseimembra monkey. PMID- 9406800 TI - Influence of different systems of feeding in the appearance of cryptosporidiosis in goat kids. AB - This study evaluated how different systems of feeding may influence the appearance, maintenance, or both of cryptosporidial infection. Animals reared with natural lactation, a traditional artificial feeding system, and a variety of the latter were studied for oocysts in feces. The diagnosis was made by examination of fecal smears stained with auramine-O. Morbidity and mortality were high, particularly in farms with bad hygienic conditions and natural feeding systems. The traditional artificial feeding system is not enough to reduce the presence of parasites. Isolation of newborns at birth and colostrum administration with a feeding bottle, obtained in the most aseptic conditions possible, seems to be an effective prophylactic method for cryptosporidiosis control. PMID- 9406801 TI - Map turtle winter leech loads. AB - Adult common map turtles, Graptemys geographica (n = 243), were obtained in November 1995 from a hibernation site in the Lamoille River, Vermont. Of the 208 female turtles examined, 151 (72.6%) had at least 1 leech (Placobdella parasitica) attached and 10 of 35 males (28.6%) were similarly parasitized. Mean abundances were 1.49 (SD = 1.461, n = 208) for female turtles and 0.34 (SD = 0.591, n = 35) for males; the difference was significant (t = 4.558, df = 241, P < 0.001). Leech broods were found on 34 of 208 female turtles (16.3%) and 2 of 35 males (5.7%). One of the leeches was of record size (77.7 mm total length); another specimen measuring 64.4 mm had 153 brood-sized (x = 4.5 mm) young attached to its venter. Because of poor visibility and partial ice cover, only 7 turtles were recovered in March 1996. All of these turtles had attached leeches, and 4 turtles had broods of 9-52 young ranging in length from 4.58 to 5.78 mm. One Placobdella ornata was found in the March sample. Our results suggest that leeches of various size classes remain attached to hibernating adult map turtles throughout the winter. PMID- 9406799 TI - Ticks, Lyme disease spirochetes, trypanosomes, and antibody to encephalitis viruses in wild birds from coastal Georgia and South Carolina. AB - Ticks and blood samples were collected from wild birds mist-netted on St. Catherine's Island, Georgia, and at the Wedge Plantation in coastal South Carolina in 1994 and 1995. Immature stages of 5 species of ixodid ticks were recovered from 10 of 148 (7%) birds belonging to 6 species in Georgia, whereas 6 ixodid species were recovered from 45 of 259 (17%) birds representing 10 avian species in South Carolina. Borrelia burgdorferi sensu lato was isolated from 27 of 120 (23%) screened ticks (Ixodes scapularis and Ixodes minor) recovered from South Carolina birds, but from none of 16 screened ticks removed from Georgia birds. This spirochete was also isolated from 1 of 97 (1%) birds in South Carolina. In 1995, neither eastern equine encephalitis (EEE) virus nor St. Louis encephalitis (SLE) virus was isolated from any of 218 bird sera screened, but serum neutralizing antibodies were found to EEE virus in 4 of 121 (3%) sera and to SLE virus in 2 of 121 (2%) sera from South Carolina. No antibody to either virus was detected in 51 avian sera screened from Georgia. Trypanosomes (probably Trypanosoma avium) were isolated from 1 of 51 (2%) birds from Georgia and from 13 of 97 (13%) birds from South Carolina. Our data suggest that some wild birds may be reservoir hosts for the Lyme disease spirochete and for encephalitis viruses in coastal Georgia and South Carolina and that migrating birds can disperse immature ticks infected with B. burgdorferi. PMID- 9406803 TI - In vitro cultivation and experimental inoculation of Sarcocystis falcatula and Sarcocystis neurona merozoites into budgerigars (Melopsittacus undulatus). AB - This is a report of the isolation and in vitro propagation of merozoites of Sarcocystis falcatula. Culture-derived S. falcatula merozoites remained infectious after several passages in tissue culture as determined by the susceptibility of experimentally inoculated Melopsittacus undulatus that died of pulmonary sarcocystosis after experimental inoculation. There was no obvious clinical disease or lesion when Sarcocystis neurona merozoites were used to inoculate M. undulatus. On the basis of the biological infectivity data, S. falcatula and S. neurona are not synonymous as suggested from limited molecular data. PMID- 9406802 TI - Marked eosinophilia in interleukin-5 transgenic mice fails to prevent Trichinella spiralis infection. AB - In order to study the role of eosinophils in the host defense against Trichinella spiralis infection, worm recovery after infection with T. spiralis was compared between interleukin-5 transgenic (IL-5 Tg) mice with a constant high level of peripheral eosinophils and nontransgenic C3H/HeN mice. No significant difference in the recovery of muscle larvae or adult worms in the small intestine, fecundity of female adult worms, or infectivity of newborn larvae was observed between nonimmunized C3H/HeN and IL-5 Tg mice or C3H/HeN and IL-5 Tg mice immunized with somatic antigen of T. spiralis. However, a significant difference was observed in the fecundity of female adult worms and recovery of muscle larvae between nonimmunized and immunized IL-5 Tg mice or C3H/HeN mice. These results demonstrate that having more eosinophils does not improve immunity against the various aspects of T. spiralis infection. PMID- 9406804 TI - Endoparasites of plethodontid salamanders from Paradise Brook, New Hampshire. AB - Totals of 52 dusky salamanders Desmognathus fuscus, 51 two-lined salamanders Eurycea bislineata, 54 red-backed salamanders Plethodon cinereus, and 3 spring salamanders Gyrinophilus porphyriticus (Plethodontidae) collected in June and August 1995 from Paradise Brook, a tributary to Hubbard Brook, New Hampshire, were examined for parasites. Parasites found were Brachycoelium storeriae, Brachycoelium sp., Bothriocephalus rarus, Falcaustra sp., Omeia sp., Batracholandros magnavulvaris, and Cepedietta michiganensis. Eighty-six percent of the red-backed salamanders, a terrestrial species, harbored 1 or more parasites. Among the aquatic and semiaquatic species, 27% of the dusky and 45% of the two-lined salamanders were infected with 1 or more parasites. PMID- 9406805 TI - Trichinella nelsoni in carnivores from the Serengeti ecosystem, Tanzania. AB - A survey of trichinellosis among sylvatic carnivore mammals from the Serengeti ecosystem (Tanzania) demonstrated the presence of Trichinella nelsoni in 5 of 9 species examined. Muscle samples were collected from carcasses of 56 carnivores from 1993 to 1995 and frozen before transport and examination. Following artificial digestion of the samples, collected larvae were analyzed by the random amplified polymorphic DNA technique. Trichinella nelsoni was identified in 1 bat eared fox (Otocyon megalotis), 1 cheetah (Acinonyx jubatus), 1 leopard (Panthera pardus), 3 lions (Panthera leo), and 3 spotted hyenas (Crocuta crocuta). The numbers of bat-eared foxes (6), cheetahs (5), and leopards (3) examined were too small to reveal the roles of these carnivore species in the ecology of T. nelsoni. The numbers of lions and spotted hyenas examined, with a prevalence of 12% and 23%, respectively, suggest that these species may be reservoirs of T. nelsoni in the area under study. PMID- 9406806 TI - Sarcocystosis in mink (Mustela vison). AB - This report describes the clinical, microscopic, and ultrastructural findings in mink with muscular sarcocystosis. Three 2-3-mo-old mink were killed because they were ill with signs of progressive neurological disease. One mink had variable numbers of sarcocysts in multiple skeletal muscles. Sarcocysts were up to 300 microm in long and 20 microm wide. Ultrastructurally, the sarcocyst wall had numerous elongated 1.7-2.0-microm x 250-nm villar protrusions (VP). The VP had microtubules and irregularly distanced minute undulations. Both metrocytes and bradyzoites were present in sarcocysts. The mink with sarcocysts in muscles also had nonsuppurative meningoencephalitis and meningomyelitis. Similar brain lesions were found in other 2 mink from the same farm, but sarcocysts were not observed in the skeletal muscle of these animals. This is the first report of muscular sarcocystosis in mink. PMID- 9406807 TI - Differential effects of sunscreens on UVB-induced immunomodulation in humans. AB - Ultraviolet radiation has been shown to suppress the (skin) immune system both in animal species and in humans. Whether sunscreens can prevent immunosuppression is a matter of debate. This study investigated the protective capacity of a commercial sunscreen lotion in humans. Part of the right arm of healthy volunteers was exposed to erythemagenic ultraviolet B doses of 160 mJ per cm2 for four consecutive days. Before irradiation, sunscreen was applied either directly onto the skin or onto a piece of quartz fixed to the skin (to avoid penetration of the sunscreen in the epidermis where it cannot block the photoisomerization of trans-urocanic acid in cis-urocanic acid in the stratum corneum). The control group was irradiated without prior application of sunscreen. Four h after the last irradiation, epidermal sheets were obtained by the suction-blister method from both arms and epidermal cells were used as stimulator cells in the mixed epidermal cell lymphocyte reaction. Responses directed to epidermal cells derived from irradiated skin were expressed as percentages of responses directed to epidermal cells derived from the nonirradiated left arm. The mixed epidermal cell lymphocyte reaction responses in the control group were found to be significantly increased (205%). This enhancement of the mixed epidermal cell lymphocyte reaction responses was associated with an influx of CD36+DR+ macrophages in the irradiated skin. Application of the sunscreen, either onto a piece of quartz or directly onto the skin, prevented the increase of the mixed epidermal cell lymphocyte reaction responses and the influx of CD36+DR+ cells. In an earlier study, volunteers were exposed three times weekly to suberythemagenic doses of ultraviolet B over 4 wk, resulting in mixed epidermal cell lymphocyte reaction responses that were decreased to 20%. The same sunscreen was not able to prevent this suppression. These contradicting results indicate that the protective effect of sunscreens with respect to ultraviolet-induced immunomodulation is critically dependent on the choice of ultraviolet treatment. PMID- 9406808 TI - UVB induction of epithelial tumors in human skin using a RAG-1 mouse xenograft model. AB - To examine the effects of chronic ultraviolet light on human epidermal cells, we grafted white human skin onto recombinase activating gene-1 knockout mice. We found previously that the maximal concentration of ultraviolet B radiation (290 320 nm) tolerated by human skin xenografts was 500 J per m2 when given three times weekly. One hundred and fifty-eight grafted mice were randomized and observed for a median of 10 mo in four groups: (i) no treatment; (ii) one treatment with the chemical carcinogen dimethyl-(a)benzanthracene; (iii) ultraviolet B three times weekly; and (iv) a combination of dimethyl (a)benzanthracene and ultraviolet B. Approximately half of the skin specimens treated with ultraviolet B developed superficial milia and epidermal cysts. Grafts contained up to seven milia lesions between 4 and 8 mo after initiation of treatment, whereas the number of larger epidermal cysts was rarely more than two. Milia and cysts developed in the skin regardless of pigmentation or tanning. Actinic keratoses arose in 9% of grafts treated with ultraviolet B alone and in 19% of grafts treated with the combination of dimethyl-(a)benzanthracene and ultraviolet B. Invasive squamous cell carcinomas developed in 10% of grafts after combined dimethyl-(a)benzanthracene and ultraviolet B treatment and lesions were restricted to skin grafts that did not tan. These findings demonstrate that (i) development of ultraviolet-induced lesions can be experimentally accelerated in human skin, (ii) xenografted recombinase activating gene-1 deficient mice are superior to severe combined immunodeficiency disease mice for chronic ultraviolet B studies, and (iii) benign cystic tumors and squamous cell carcinomas are caused by ultraviolet B. PMID- 9406809 TI - Adhesion of leukocytes to dermal endothelial cells is induced after single-dose, but reduced after repeated doses of UVA. AB - Approximately 20-50% of ultraviolet A (UVA) irradiation delivered to the skin surface may reach the human dermal microvascular endothelial cells (HDMEC) that play a pivotal role in cellular inflammatory tissue; however, the pathophysiologic role of HDMEC in UVA-induced skin changes is largely unknown. Based on previous in vivo and in vitro studies revealing UVA-induced expression of endothelial adhesion molecules, we studied isolated HDMEC under various conditions in order to further delineate the impact of UVA on these cells. The expression of cell adhesion molecules was determined by flow cytometry and the resulting changes of stable adhesion of leukocytes to endothelial cells were quantitated for granulocytes, lymphocytes, and monocytes using a newly developed multicellular adhesion assay. Additionally, antibody blocking experiments were performed to delineate the role of individual cell adhesion molecules in UVA induced leukocyte adherence. High-dose polychromatic UVA (25 J per cm2, maximal emission at 375 nm) induced intercellular adhesion molecule-1 and E-selectin with different kinetics but correlating the adhesion of leukocyte subsets. This effect subsided, however, in the course of 3-6 daily applied UVA doses. Moreover, pro inflammatory cytokine challenge by tumor necrosis factor-alpha and interleukin-1 alpha resulted in significantly weaker induction of intercellular adhesion molecule-1 and E-selectin in repeatedly UVA-exposed HDMEC. Differential quantitation of peripheral blood derived granulocytes, lymphocytes, and monocytes revealed reduced adhesion particularly of lymphocytes followed by monocytes and granulocytes compared with leukocyte adhesion to nonirradiated but cytokine stimulated HDMEC. It is concluded that UVA substantially influences endothelial cell adhesion molecules expression and thus directly interferes with leukocyte adhesion to endothelial cells. Divergent UVA-induced effects in this respect can be attributed to the mode of UV exposure as well as to the condition of endothelial cells prior to UVA exposure. PMID- 9406810 TI - Genetic variation in low-dose UV-induced suppression of contact hypersensitivity and in the skin photocarcinogenesis response. AB - Two of the major cutaneous consequences of ultraviolet (UV) radiation exposure are immunosuppression and the development of skin cancer. This study examined whether these effects are genetically determined. Suppression of contact hypersensitivity by local, low-dose UV radiation was examined in what have been termed "UV-susceptible" and "UV-resistant" strains of mice. C3H/HeJ mice ("UV resistant") were resistant to the adverse effects of low-dose UV radiation when normal doses of hapten were applied to UV-irradiated skin; however, they were sensitive when the amount of hapten used for sensitization was reduced. A similar effect was observed in BALB/c mice ("UV resistant") and when the hapten was dimethylbenz(a)anthracene, thus indicating that the genetic variation was not strain or hapten specific. Despite the fact that some strains were sensitive and some were resistant to low-dose UV radiation when high doses of hapten were employed, all strains initially sensitized to hapten through UV-irradiated skin were found to be unresponsive when rechallenged on normal skin, no matter what the initial sensitizing dose of hapten was. To determine whether other biologic effects of UV also exhibited genetic variation, C3H/HeN and C3H/HeJ mice were compared for susceptibility to UVB-induced skin cancer formation. C3H/HeJ mice developed significantly more tumors than C3H/HeN mice when subjected to a single dose of UV radiation followed by repeated exposure to the tumor promoter 12-O tetradecanoyl-phorbol-13-acetate. These studies provide strong evidence that genetic factors influence individual susceptibility to the biologic effects of UV radiation. PMID- 9406811 TI - Role of p53 in UVB-induced apoptosis in human HaCaT keratinocytes. AB - Apoptosis represents an active form of cell death that is involved in the control of tissue homeostasis and in the deletion of DNA-damaged cells. Because the product of the tumor suppressor gene p53 has been demonstrated to be crucial for the induction of apoptosis in certain cell types, the present study was aimed at elucidating its role in ultraviolet-induced apoptosis in HaCaT keratinocytes. After in vitro ultraviolet B irradiation, p53 protein levels were noted to increase prior to the induction of apoptosis in a time- and concentration dependent fashion. This increase could not be inhibited by the protein synthesis inhibitor cycloheximide. Because HaCaT keratinocytes are known to bear two p53 point mutations and because it is unclear whether p53 in HaCaT cells is still functional regarding induction of apoptosis, HaCaT cells were stably transfected with wild-type p53 cDNA inserted into the expression vector pCMV-Neo-Bam in sense (pC53-SN3) and anti-sense (pC53-ASN) direction. After selection with geniticin, growing colonies were screened for the presence of the transfected cDNA constructs by polymerase chain reaction. Cell clones bearing the anti-sense product were further analyzed for p53 expression by western blotting. Clones showing reduced p53 protein levels were irradiated with ultraviolet B light, and there was a clear reduction of apoptosis in the pC53-ASN bearing cell clones compared with the parental HaCaT cells. These studies demonstrate that blocking mutated p53 can partially block apoptosis in HaCaT keratinocytes and furthermore can confirm the key role for p53 in ultraviolet-induced apoptosis in human keratinocytes. Moreover, HaCaT keratinocytes and their p53-transfectants provide a convenient model that allows for further detailed analyses of apoptosis associated biochemical and molecular events in human keratinocytes. PMID- 9406812 TI - Expression of B7-1 by Pam 212 squamous cell carcinoma enhances tumor cell interactions with dendritic epidermal cells but does not affect in vivo tumor growth. AB - Direct antigen presentation of tumor-associated antigens by tumor cells to T lymphocytes may induce clonal anergy as a mechanism of escape from immune surveillance. B7-1 is a costimulatory molecule for the activation of both CD4+ and CD8+ T lymphocytes that prevents the induction of clonal anergy. Thus, the transfer of B7-1 genes into tumor cells can induce protective immunity and lead to tumor rejection of some tumors in model systems of in vivo tumor growth; however, there is no information on whether stable expression of B7-1 can affect the in vivo growth of squamous cell carcinoma, a common skin cancer. Here, we study how the stable cell surface expression of high levels of B7-1 by Pam 212, a murine squamous cell carcinoma, affects tumor cell-lymphocyte interactions (lymphocyte proliferation and cytotoxicity). Consistent with its costimulatory role, we demonstrate that B7-1 can efficiently induce dendritic epidermal T-cell proliferation in three different dendritic epidermal T-cell cell lines. In addition, B7-1 enhances dendritic epidermal T-cell cytolytic activity against Pam 212 cells in an in vitro 51Cr-release assay, which was blocked by CTLA-4/Ig fusion protein. In contrast to dendritic epidermal T cells, the expression of B7 1 does not alter Pam 212 interactions with either cytotoxic T-lymphocytes, natural killer, or lymphokine-activated killer cells. B7-1 expression by Pam 212 cells did not alter its ability to grow tumors in vivo, as their rate of tumor growth was the same as vector-transfected Pam 212 cells, which were B7-1 negative. Our studies indicate that B7-1 gene transfer into Pam 212 does not alter its tumorigenicity, because it does not alter tumor cell-lymphocyte interactions with cytotoxic T lymphocytes, natural killer cells, and lymphokine activated killer cells. Further studies of B7-1 modified Pam 212 and dendritic epidermal T cells will clarify whether T-cell receptor-gamma/delta-bearing T lymphocytes can play a role in immunotherapy of Pam 212 squamous cell carcinoma. PMID- 9406813 TI - T lymphocytes from a subset of patients with pemphigus vulgaris respond to both desmoglein-3 and desmoglein-1. AB - Pemphigus vulgaris and pemphigus foliaceus are cutaneous autoimmune diseases characterized by intraepithelial blisters and autoantibodies to desmosomal glycoproteins. The antigens recognized by pemphigus vulgaris and pemphigus foliaceus autoantibodies are desmoglein-3 (Dsg3) and desmoglein-1 (Dsg1), respectively. Dsg3 and Dsg1 are members of the desmoglein subfamily of the cadherin supergene family of cell adhesion molecules. It has been well documented that a subset of pemphigus vulgaris sera have IgG reactivity to both Dsg1 and Dsg3, suggesting that Dsg1 may also participate in the autoimmune response of these patients. The cellular mechanisms of T cell autoimmunity in these patients, however, are completely unknown. In this study, we tested the proliferative responses of T lymphocytes from eight pemphigus vulgaris patients after incubation with Dsg3 and Dsg1 fusion proteins. The sera of four of these PV patients showed reactivity with both Dsg1 and Dsg3, whereas the remaining four reacted only with Dsg3. We found that T cells obtained from those patients that exhibited the combined Dsg1/Dsg3 autoantibody reactivity showed a proliferative response after exposure to either Dsg1 or Dsg3 fusion proteins. The cellular responses to both of these recombinant proteins were highly specific and restricted to the CD4-positive T cell population. T cells from pemphigus vulgaris patients with no anti-Dsg1 serum reactivity showed a proliferative response to Dsg3, but not to Dsg1. The Dsg1 fusion protein used in this study has minimal sequence homology with Dsg3. Thus, this study provides the first evidence that T cells from a subset of pemphigus vulgaris patients respond to both Dsg1 and Dsg3. PMID- 9406814 TI - Fibronectin upregulates in vitro generation of dendritic Langerhans cells from human cord blood CD34+ progenitors. AB - Several studies have demonstrated that dendritic cells can be generated in vitro from CD34+ hematopoietic progenitor cells. In vivo, dendritic cells are found in many tissues and reside in direct proximity to extracellular matrix proteins. Because extracellular matrix proteins affect differentiation and location of cells in tissues, this study was designed to investigate potential effects of extracellular matrix proteins on differentiation of dendritic cells. Dendritic cells were generated from CD34+ human cord blood cells in the presence of granulocyte-macrophage colony-stimulating factor and tumor necrosis factor-alpha for 6 d and subsequently cultured for an additional 6-d period on tissue culture plates coated with various extracellular matrix proteins. Among the extracellular matrix proteins tested, exposure to fibronectin stimulated dendritic cell/Langerhans cell differentiation as indicated by the 50% increase of the number of cells expressing the Birbeck granule-associated marker Lag and displaying numerous Birbeck granules. Adhesion on fibronectin was shown to be specifically mediated by the integrin alpha5beta1. Because laminin and collagen were unable to cause similar changes in Langerhans cell development, these results suggest that fibronectin may cause changes affecting cellular differentiation of progenitors. Hematopoietic progenitors may exhibit maturational regulated differences in response to both matrix molecules and cytokines. The influence of combined signals emanating from a supportive microenvironment, specific integrins, and particular cytokines in the differentiation of Langerhans cells is discussed. PMID- 9406815 TI - Transformed and nontransformed human T lymphocytes migrate to skin in a chimeric human skin/SCID mouse model. AB - To study human T cell migration to human skin in vivo, we grafted severe combined immunodeficient mice with 500-microm thick human skin. Two weeks after grafting, epidermal and dermal structures in the grafts were of human origin. When we intraperitoneally injected grafted mice with clones of the human HUT-78 T cell line derived from a patient with cutaneous T cell lymphoma and Sezary syndrome, we detected in the grafts the rare Vbeta23-Jbeta1.2 T cell receptor transcripts characteristic for the HUT-78 clones. These signals were found 2-6 d after cell injection in about 40% of the grafted and HUT-78 cell injected mice but not in grafts from mice that received no exogenous T cells. In contrast to HUT-78 cells, which only accumulate in low number, grafts topically challenged with nickel sufate in vaseline from mice that were injected with autologous nickel-reactive T cell lines led to massive accumulation of T cells within 3 d. Only scattered T cells accumulated in the skin when grafted mice received vaseline plus T cells, nickel sulfate alone, T cells alone, or nickel sulfate plus an allogeneic nickel nonreactive T cell clone. When the T cell lines were labeled with the fluorochrome PKH-26 before cell injection, spots of fluorescent label in the size and shape of cells were found in the grafts challenged with nickel. Together, these results clearly demonstrate that human T cells can migrate to human skin in this chimeric human/mouse model. PMID- 9406816 TI - EGF-receptor tyrosine kinase inhibition induces keratinocyte growth arrest and terminal differentiation. AB - Epidermal keratinocyte growth and differentiation are regulated by specific families of growth factors and receptors. Peptide growth factors of the epidermal growth factor family stimulate proliferation of clonal density human keratinocytes and suppress markers of terminal differentiation in confluent cultures of human keratinocytes. We present evidence that selected inhibitors of activation of the type I human epidermal growth factor receptor (EGFR or HER-1), namely, neutralizing monoclonal antibody to HER-1/EGFR and the specific tyrosine kinase inhibitor PD 153035, potently inhibit proliferation of human keratinocytes in autonomously replicating subconfluent cultures. Coupled to growth arrest is the suppression of HER-1 tyrosine autophosphorylation in inhibitor-treated human keratinocytes. Proliferation and tyrosine autophosphorylation are initially reversible following removal of the inhibitor and restimulation of cells with epidermal growth factor. Sustained inactivation of HER-1 in autonomously replicating cultures of human keratinocytes induces expression of keratin 1 and keratin 10 genes, early markers of terminal differentiation. Reversal of growth inhibition by epidermal growth factor suppresses keratin 1 and keratin 10 expression. These results demonstrate that human keratinocyte terminal differentiation as well as proliferation are mediated by HER-1. Co-expression of autocrine epidermal growth factor-related ligands as well as HER-1 by human keratinocyte may function as part of the signal transduction network in epidermis to regulate cell number, replication rate, and terminal differentiation. PMID- 9406817 TI - Autocrine nerve growth factor protects human keratinocytes from apoptosis through its high affinity receptor (TRK): a role for BCL-2. AB - Normal human keratinocytes synthesize and release nerve growth factor (NGF) and express both the low- and the high-affinity NGF receptor. Because NGF has been shown to rescue certain cell types from programmed cell death, we investigated the role of endogenous NGF in preventing keratinocyte apoptosis. We report here that apoptosis is induced in normal human keratinocytes in culture by blocking endogenous NGF signaling with either anti-NGF neutralizing antibody or K252, a specific inhibitor of the tyrosine kinase high-affinity NGF receptor. Apoptosis was assessed by DNA laddering, electron microscopy, and in situ nick end labeling technique. In anti-NGF-treated keratinocytes, the apoptotic process starts at 96 h, and is maximal at 120 h. After K252 treatment, apoptosis starts at 48 h and peaks at 120 h. Because the product of the bcl-2 proto-oncogene protects many cell types from apoptosis, we measured the levels of this protein in apoptotic keratinocytes. We found that both K252 and anti-NGF antibody strikingly downregulate bcl-2 expression, starting at 72 h. Furthermore, HaCat keratinocytes stably transfected with a plasmid containing bcl-2 cDNA fail to undergo apoptosis when treated with K252. These findings show that autocrine NGF acts as a survival factor for human keratinocytes in vitro through its high-affinity NGF receptor, possibly by maintaining constant levels of Bcl-2. PMID- 9406818 TI - Intracellular calcium oscillations in cell populations of ras-transfected I-7 subline of human HaCaT keratinocytes. AB - We have observed oscillations of intracellular Ca2+ (Ca[i]) concentration in populations of ras-transfected HaCaT keratinocytes of I-7 subline. In postconfluent monolayers of I-7 keratinocytes, an increase in extracellular Ca2+ (Ca[o]) concentration to 0.25-0.5 mM induced sinusoidal Ca(i) oscillations, which persisted longer than 1 h with amplitudes of 50-150 nM and periods of 5-10 min. Thapsigargin, which depletes internal Ca2+ stores, did not prevent Ca(o)-induced Ca(i) oscillations, and it also induced Ca(i) oscillations in the ras-transfected I-7 line. Removal of extracellular Ca2+ or addition of Ca2+-entry blocker La3+ or SK&F 96365 inhibited Ca(i) oscillations, suggesting that Ca(i) oscillations in ras-transfected HaCaT keratinocytes were dependent on Ca2+ influx across the plasma membrane. Because the Ca(o)-induced Ca(i) oscillations have been observed only in ras-transfected I-7 subline and not in its nontransfected parental HaCaT line, this may provide a partial explanation for the divergent responses of ras transfected and nontransfected keratinocytes to Ca(o) signal for control of growth and differentiation. PMID- 9406819 TI - The role of IGF-I in human skin and its appendages: morphogen as well as mitogen? AB - Previous studies have investigated the expression of insulin-like growth factor-I (IGF-I) and its receptor in cultured skin cells or in whole skin. In order to fully understand the role of IGF-I in the skin and its appendages, however, a comprehensive study that details the expression of IGF-I and the IGF-I receptor in sections of human skin is needed. Therefore, we now report an immunocytochemical and in situ hybridization localization study of the cell types expressing IGF-I and its receptor in human adult skin and its appendages. We have observed that (i) dermal fibroblasts produce IGF-I, (ii) the epidermal basal keratinocytes are IGF-I negative but IGF-I receptor positive, and (iii) the keratinocytes of the stratum granulosum produce IGF-I. These observations indicate either that the mitogenesis of the basal keratinocytes is regulated by IGF-I expressed both in the dermis and in the stratum granulosum, or that dermal fibroblasts are responsible for sequestering IGF-I to the basal keratinocytes and that the stratum granulosum-derived IGF-I may be an autocrine regulator of epidermal differentiation. The distribution of IGF-I and its receptor in the hair follicle indicates that IGF-I may be a morphogen, not a mitogen, at those sites, because their proliferating cells, but not their differentiating cells, are IGF-I receptor negative. Further, IGF-I receptor expression by the dermal papilla appears to be switched off during the transition from anagen to catagen, which implies a regulatory role for IGF-I during the hair growth cycle. PMID- 9406820 TI - A transglutaminase-related antigen associates with keratin filaments in some mouse epidermal cells. AB - A mouse monoclonal IgG, G82, directed against guinea pig liver transglutaminase recognizes a transglutaminase-related antigen that is associated with the keratin intermediate filament network in some primary mouse keratinocytes. The association can be seen at the resolution of individual keratin tonofibrils following fixation and staining for double-label indirect immunofluorescence. Western blots indicate that G82 reacts with two proteins of 95 kDa and 280 kDa, respectively, in extracts of these cells. The 95-kDa band is also recognized by a polyclonal antibody against purified guinea pig liver transglutaminase, and the 280-kDa protein seems to correspond to a similar protein that was shown to be recognized by G92.1.2 in the intermediate filament fraction of primary mouse fibroblasts. The transglutaminase-related antigen was shown by confocal microscopy to co-localize only with nonbasal cell specific keratin intermediate filaments. PMID- 9406821 TI - Permeability barrier disruption coordinately regulates mRNA levels for key enzymes of cholesterol, fatty acid, and ceramide synthesis in the epidermis. AB - The extracellular lipids of the stratum corneum, which are comprised mainly of cholesterol, fatty acids, and ceramides, are essential for epidermal permeability barrier function. Moreover, disruption of the permeability barrier results in an increased cholesterol, fatty acid, and ceramide synthesis in the underlying epidermis. This increase in lipid synthesis has been shown previously to be due to increased activities of HMG-CoA reductase, acetyl-CoA carboxylase, fatty acid synthase and serine palmitoyl transferase, key enzymes of cholesterol, fatty acid, and ceramide synthesis, respectively. In the present study, we determined whether the mRNA levels for the key enzymes required for synthesis of these three classes of lipids increase coordinately during barrier recovery. By northern blotting, the steady-state mRNA levels for HMG-CoA reductase, HMG-CoA synthase, farnesyl pyrophosphate synthase, and squalene synthase, key enzymes for cholesterol synthesis, all increased significantly after barrier disruption by either acetone or tape stripping. Additionally, the steady-state mRNA levels of acetyl-CoA carboxylase and fatty acid synthase, required for fatty acid synthesis, as well as serine palmitoyl transferase, the rate-limiting enzyme of de novo ceramide synthesis, also increased. Furthermore, artificial restoration of the permeability barrier by occlusion after barrier disruption prevented the increase in mRNA levels for all of these enzymes, except farnesyl pyrophosphate synthase, indicating a specific link of the increase in mRNA levels to barrier requirements. The parallel increase in epidermal mRNA levels for the enzymes required for cholesterol, fatty acid, and ceramide synthesis may be due to one or more transcription factors that regulate lipid requirements for permeability barrier function in keratinocytes. PMID- 9406823 TI - Melanogenesis in cultured melanocytes can be substantially influenced by L tyrosine and L-cysteine. AB - We investigated the effect of varying concentration of 1-tyrosine and 1-cysteine in culture medium on melanin production by human skin melanocytes (skin phototype II/III). In addition to the analyses of dopa oxidase activity and total melanin, pheomelanin production in the cells was assessed by high-performance liquid chromatography determinations of pheomelanin degradation products, 3 aminotyrosine and 4-amino-3-hydroxyphenylalanine. As another marker for pheomelanin, melanosomal sulfur was determined by the use of X-ray microanalysis. With varying concentration of both amino acids, profound changes in the pigmentation patterns of the melanocytes were observed. A high concentration of 1 tyrosine (0.2 mM) was always connected with increased pigmentation. In combination with a low 1-cysteine content we saw an increase in tyrosinase activity and the highest melanin content. At high concentrations of both 1 tyrosine and 1-cysteine, the melanocytes showed reduced tyrosinase activity and they produced notably more pheomelanin. In case of the pheomelanin measurements by high-performance liquid chromatography and the sulfur detection with X-ray microanalysis, strongly increased concentrations were found when cells were maintained in high 1-tyrosine medium as compared with those grown with low 1 tyrosine. This was especially true for the combination with low 1-cysteine showing that the 1-tyrosine content of the medium strongly influences not only the eumelanin but also the pheomelanin production in the cultured melanocyte. It can be concluded that variations in the concentrations of 1-tyrosine and 1 cysteine in culture medium can be used to regulate the melanogenetic phenotype under in vitro conditions. PMID- 9406824 TI - Deletion mapping of chromosome 3p and 13q and preliminary analysis of the FHIT gene in human nonmelanoma skin cancer. AB - Loss of heterozygosity of chromosomes 3p and 13q occurs frequently in human cutaneous squamous cell neoplasms, suggesting the presence of one or more tumor suppressor genes on these chromosome arms that may be involved in the pathogenesis of this tumor type. To date there is no clear evidence in cutaneous tumors where these putative genes are located. In this study we have analyzed 20 squamous cell neoplasms that show allelic loss at chromosome 13q, and 22 squamous cell neoplasms that show allelic loss at chromosome 3p, in an attempt to define the smallest area of deletion. One commonly deleted region was identified on chromosome 13 that centred around 13q13, and two commonly deleted regions were identified on chromosome 3 that mapped to 3p24-pter and 3p12-p14.1. Our findings suggest the presence of at least one tumor suppressor gene on chromosome 13 and two tumor suppressor genes on chromosome 3p that may be involved in the progression of these neoplasms. Deletions within the Fragile Histidine Triad gene, located at 3p14.2, have been reported in several tumors, leading to the suggestion that this gene is involved in tumor development. To evaluate the role of the Fragile Histidine Triad gene in nonmelanoma skin cancer, we have used reverse transcriptase polymerase chain reaction analysis to screen for deletions in 16 tumors (five basal cell carcinomas, five squamous cell carcinomas, five actinic keratoses, and one case of Bowen's disease) and HaCaT and A431 cell lines. A normal transcript was found to be expressed in 14 of 16 tumors and both cell lines. This suggests that the Fragile Histidine Triad gene is not a common target for deletion in Bowen's disease and the cell lines HaCaT and A431. PMID- 9406822 TI - Sorting and secretion of a melanosome membrane protein, gp75/TRP1. AB - The melanosome is an organelle specialized for melanin synthesis that is derived from the endocytic pathway. Several melanosome membrane proteins have been identified, forming a family of proteins known as tyrosinase-related proteins. Two members of this family, tyrosinase and gp75, are well-characterized melanocyte differentiation antigens. Our previous studies have shown that gp75, the mouse brown locus protein, is sorted to melanosomes along the endocytic pathway, directed by a hexapeptide sorting signal located in the cytoplasmic tail. In this study, we report the unexpected finding that a portion of gp75 is secreted. Substantial levels of secretory gp75 were detected in melanocytic cells. Cell surface expression of gp75 was also detected, representing 2% of cellular gp75. Characterization of secretory gp75 cells showed that it is: (i) a truncated form that lacks the transmembrane region, the cytoplasmic tail where the endosomal sorting signal is located, and a small portion of the lumenal domain; (ii) more extensively glycosylated than endocytic/melanosomal gp75, containing trans-Golgi processed sugar residues; and (iii) generated post translationally in an acid sensitive compartment after processing in the trans Golgi, and secreted rapidly after generation. Thus, these endocytic/melanosomal membrane proteins can be processed to abundant secretory forms, probably in an endocytic compartment through a potentially novel secretory pathway. PMID- 9406825 TI - Epidermal remodeling in psoriasis (II): a quantitative analysis of the epidermal architecture. AB - Hyperproliferative psoriatic epidermis was quantitatively analyzed using a geometric model of viable epidermis. Our model was based on hexagonally arranged cylindrical papillae, which allowed the determination of the total volume of the viable epidermis and the total area of the interface with the proliferative compartment based on several parameters, such as papillary height, papillary width, and distance between neighboring papillae. The analysis assumed that the total number of viable epidermal cells paralleled the proliferative compartment in a steady state of cell flow, so a quantitative relation could be made between both volume and interface of the viable epidermis. Multiple parameters of the psoriatic epidermal architecture were measured, and variations within psoriasis were predicted by the model. The results predicted were remarkably close to the observed values. The geometric model also indicated that psoriatic epidermis could be subdivided into two distinct types, with and without a granular layer; the latter having a shorter turnover time. This is consistent with the notion that the typical psoriatic epidermis (without the granular layer) represents the expanding hyperproliferative phase, whereas the psoriatic epidermis with a granular layer represents stationary or resolving states. The model of hexagonally arranged cylindrical papillae suggested that the architecture of the psoriatic epidermis is constructed by a simple mechanism, whereby the psoriatic angulated rete-papilla pattern was produced by a two-dimensional increase in the proliferative compartment and a three-dimensional increase in the total volume of the viable epidermis. PMID- 9406826 TI - Genetic basis of dominantly inherited transient bullous dermolysis of the newborn: a splice site mutation in the type VII collagen gene. AB - Transient bullous dermolysis of the newborn (TBDN) is a blistering disease evident at birth or shortly thereafter, but the blistering tendency decreases with advancing age. The tissue separation in TBDN is below the lamina densa, and electron microscopy has revealed abnormalities in anchoring fibrils. Immunofluorescence staining demonstrates intracellular accumulation of type VII collagen. In this study, we report a G-to-C transversion mutation in the last nucleotide of intron 35 of the type VII collagen gene (COL7A1) in a family with autosomal dominant TBDN in three generations. This nucleotide substitution abolishes the obligatory consensus 3'-acceptor splice site, predicting in-frame skipping of exon 36. Thus, TBDN in this family is caused by a mutation in COL7A1, and is therefore allelic with other variants of dominant dystrophic epidermolysis bullosa. PMID- 9406827 TI - A novel mutation in the helix termination peptide of keratin 5 causing epidermolysis bullosa simplex Dowling-Meara. AB - Epidermolysis bullosa simplex Dowling-Meara (MIM# 1317600) is the most severe of the three common epidermolysis bullosa simplex subtypes. In addition to the palmoplantar distribution seen in other epidermolysis bullosa simplex subtypes, extensive herpetiform blistering spontaneously develops on the trunk and limbs and may lead to scarring or milia formation. The keratin 5 and keratin 14 genes encode proteins that form the primary structural components of the basal epidermal keratinocytes, mutations in either of these genes can cause epidermolysis bullosa simplex. In this study we sequenced these genes in a family with epidermolysis bullosa simplex Dowling-Meara. We report a novel T to C transition in the helix termination peptide of K5 that causes a nonconservative substitution of a highly conserved amino acid within this critical region (I466T). This mutation adds to those previously reported and provides further evidence of phenotype-genotype correlation in epidermolysis bullosa simplex. PMID- 9406828 TI - Absence of HTLV-1 proviral sequences in patients with lymphomatoid papulosis. PMID- 9406830 TI - Breast cancer risk lower in women who undergo oophorectomy. PMID- 9406831 TI - Fetal laceration injury underreported. PMID- 9406829 TI - AIDS rates increase among women during first half of 90s--greatest increase found to occur through heterosexual contact. PMID- 9406832 TI - Breast disease: a primer on diagnosis and management. AB - Currently, mammography is the only method of detecting nonpalpable, early breast cancer. At this stage, 90% of the cancers are curable. Clearly, this fundamental tenet accentuates the importance of compliance and knowledge of guidelines. Although risks of mammography are minimal to nil, interpretation occasionally can be challenging, with equivocal results. New technologies are being evaluated and advances in measurement of cellular electrical potential differentials in breast tissue have produced exciting results, when compared with mammography and ultrasound. These screening efforts have increased the diagnosis of both invasive and noninvasive ductal and lobular carcinoma of the breast. For DCIS in particular, conservative, contemporary treatment options exist. These include lumpectomy with breast irradiation excluding axillary dissection. Selected patients may be treated with only lumpectomy. Although breast carcinoma is a major focus due to incidence, morbidity and mortality, the varieties of benign conditions cause many women genuine concern. Treatment options for fibrocystic change run a gamut, including cost-effective basic dietary changes, vitamin use, "health"/natural type treatments, analgesic, as well as hormonal manipulations and, on occasion, surgical intervention. Fortunately, with most patients, common sense and conservatism prevail. The presence of fibroadenomas diagnosed clinically, by ultrasound or mammography, in women aged 18-25 and beyond can create perplexing diagnostic dilemmas. Should the lesion be removed or observed? Differences of opinion exist and must be tempered by recent observations that women with complex fibroadenomas, sclerosing adenosis, epithelial calcification or papillary appocrine changes have a two- to threefold increased risk of breast cancer. The key to management in all these clinical situations is individualization. Conservatism is particularly acceptable in women under the age of 25 if a fibroadenoma is not increasing in size or not psychologically disturbing. Provoked or unprovoked nipple discharge is a clinical conundrum for patients. It is unsuspected and unwanted. While some whitish discharges result from stimulation or medication, others may have a more subtle etiology. Serous, serosanguineous, or bloody discharges mandate evaluation. Duct injection mammography and frequent excision of ductal systems are necessary. The clinician cannot forget other less common conditions, such as thrombophlebitis, fat necrosis, or infection. All clinical conditions of the breast provide a constellation of diagnostic and management problems. They are of real concern for every woman and must be resolved in an appropriate, prompt, and conscientious fashion. PMID- 9406833 TI - Uterine closure with the endo stitch 10-mm laparoscopic suturing device--a review of 50 laparoscopic myomectomies. AB - OBJECTIVE: To evaluate the mechanical performance of the Endo Stitch Laparoscopic Suturing Device and the clinical effectiveness of both a running, locked suture technique and a new modified suture technique for closure of uterine defects after laparoscopic removal of myomas. STUDY SUBJECTS: Fifty consecutive patients with symptomatic uterine leiomyomata. OBSERVATIONAL METHOD: Retrospective chart review. MAIN FINDINGS: The endometrial cavity was entered and sutured laparoscopically, in two layers, in 22 patients. In 28 patients, only the myometrium was sutured. A two-layered closure of the endometrium and myometrium was completed in an average time of 10 minutes. Mechanical problems with the Endo Stitch occurred in 11 cases. In all patients with second-look laparoscopies, the fallopian tubes were patent bilaterally without adhesions. No uterine fistulas were present in any patients with second-look laparoscopies. Posterior myomas were removed and sutured without adhesion formation. Grade 3 adhesions, to the uterine surface, were associated with transverse incisions of the uterus and over treatment with GnRH analogs. CONCLUSIONS: The Endo Stitch Laparoscopic Suturing Device in combination with a running, locked suture technique achieves a rapid, hemostatic, clinically secure closure of the endometrium and myometrium. The Endo Stitch and our modified suture technique were not associated with adhesions or blockage of the fallopian tubes or uterine fistulas following laparoscopic myomectomies. The initial mechanical problems with the Endo Stitch were resolved. In our experience, currently the Endo Stitch is the best instrument for laparoscopic suture closure of uterine defects. PMID- 9406834 TI - Interleukin-2 production by cultured human granulosa cells. AB - OBJECTIVE: To determine whether cultured human granulosa cells (GC) produce Interleukin (IL)-2 and whether this GC IL-2 production may be regulated by human chorionic gonadotropin (hCG). MATERIALS AND METHODS: Human GC derived from preovulatory follicles during in vitro fertilization (IVF) cycles were cultured in the absence or presence of hCG. Interleukin-2 was measured in tissue-culture medium by an enzyme-linked immunosorbent assay (ELISA) system. RESULTS: Cultured GCs were demonstrated to produce IL-2 in vitro. Moreover, a positive, but not statistically significant, dose-response effect was demonstrated between culture supernatant IL-2 levels and hCG concentration. CONCLUSIONS: Our data indicate that human GC produce IL-2 and that this production might be regulated by hCG. These results lend credence to the theory that IL-2 might be involved in the events leading to ovarian hyperstimulation syndrome (OHSS). PMID- 9406835 TI - Treatment of idiopathic oligozoospermia with an alpha-blocker: a placebo controlled double-blind trial. AB - OBJECTIVE: To evaluate the efficacy of terazosin, an alpha-blocker, for the treatment of idiopathic oligozoospermia. PATIENTS AND METHODS: Thirty couples with infertility whose only detectable abnormality was male idiopathic subfertility entered the study. The diagnosis of idiopathic subfertility in all males studied, aged 26 to 38 years (mean 28.2 years), was confirmed after exclusion of any iatrogenic, systemic, congenital, infectious, autoimmune or endocrinological cause. In order to start with a baseline value before the study, at least three semen samples were evaluated in accordance with the WHO recommendation. Before initiation of treatment, blood samples were drawn for measurement of FSH, LH, testosterone, prolactin, dihydrotestosterone, and estradiol. Fifteen randomly selected patients (Group A) received 2 mg/d of alpha blocker (terazosin), while another 15 (Group B) were administered an identically packed placebo tablet. Both groups received therapy for 6 months. RESULTS: The mean seminal volume changed insignificantly between the two groups (4.15 +/- 1.95 vs. 4.10 +/- 1.95). There was a statistically significant increase of the sperm concentration in patients who received the alpha-blocker compared to those receiving placebo (24.76 +/- 9.45 vs. 13.15 +/- 11.55 millions/mL; P < .001). No improvement of the mean percentage of abnormal spermatozoa was observed in the treated patients, nor a statistically significant difference of sperm motility in the treated group compared to the placebo group. Side effects were not observed in the patients receiving terazosin treatment, or were so minimal that therapy was continued. The pregnancy rates did not differ between the two groups to a statistically significant degree. CONCLUSION: The administration of terazosin to patients with idiopathic oligozoospermia has a demonstrably positive effect, especially on sperm concentration. PMID- 9406836 TI - Follicular fluid hormone concentrations after ovarian stimulation using gonadotropin preparations with different FSH/LH ratios. I. Comparison of an FSH dominant and a purified FSH preparation. AB - OBJECTIVE: A small amount of LH is necessary for 17beta-estradiol production in the ovarian follicle. Human menopausal gonadotropin (hMG) contains equal amounts of FSH and LH activity, whereas recombinant FSH is a gonadotropin preparation without LH. The aim of the present randomized study was to investigate whether ovarian stimulation treatment with recombinant FSH or hMG resulted in different steroidal composition of follicular fluid. METHODS: Antral fluid from mature follicles was collected in in vitro fertilization cycles and concentrations of testosterone, androstenedione, estrone, estradiol, progesterone, FSH, and LH were determined. Seven patients (27 samples) were treated with hMG, 6 patients (22 samples) with recombinant FSH. RESULTS: Androgen, estrogen, progesterone, and FSH concentrations in follicular fluid tended to be lower in the group treated with recombinant FSH, but the variation was large and differences were statistically not significant. CONCLUSION: Treatment with a gonadotropin preparation containing no LH resulted in adequate androgen and estrogen levels in antral fluid of the ovarian follicle in women with normal endocrine profiles, even during pituitary suppression by a GnRH agonist. Apparently, the amount of endogenous LH was sufficient for steroid production within the follicle. PMID- 9406837 TI - Thermoregulated radiofrequency endometrial ablation. AB - OBJECTIVE: To test the hypothesis that treating dysfunctional uterine bleeding by automated application of electrothermal energy to the uterine cavity, with precise regional control, might yield results equivalent to those reported for hysteroscopically directed laser and electrosurgical endometrial ablations. MATERIALS AND METHODS: Patients with life style compromising menorrhagia, referred to six gynecologic surgical centers for hysterectomy or endometrial ablation, were admitted to the study if they had normal cervical cytology, a benign endometrial biopsy, no defined cause for their bleeding, and consented to participate in the evaluation of a newly developed Vesta DUB Treatment System. The device consists of a silicone-inflatable electrode carrier to be inserted into the uterine cavity and a controller to monitor and distribute current from a matched electrosurgical generator. Treatment involved a 3-minute or shorter warm up period and a 4-minute treatment phase. RESULTS: Three- to 24-month follow-up data were available for 187 patients, with a mean follow-up of 14.8 months. The amenorrhea rate was 38%. Bleeding was reduced in 95% of patients. Actuarially, 88 +/- 3% of patients should expect to be free of menorrhagia, dissatisfaction, or need for a second procedure out to 24 months. CONCLUSIONS: The unique regional feed-back control offered by this system causes thorough, evenly distributed, thermal destruction 4-5 mm into the myometrium that reduces bleeding with durability equivalent to published reports of hysteroscopic endometrial ablation. PMID- 9406838 TI - Chondrogenic cell differentiation from membrane bone periostea. AB - Most craniofacial membrane bones are derived from neural crest (NC) cells. Interaction between NC cells and epithelium, and cellular condensation, are two major events that lead NC cells to become osteoblasts that deposit membrane bone. Unlike endochondral bone, membrane bone formation is not preceded by cartilage formation in normal development. However, chondrogenic potential in membrane bone is evidenced by several cartilage-associated phenomena in vivo. Furthermore, in vitro, periosteal cells of some membrane bones express cartilage phenotype gene products and even differentiate into chondrocytes. Hence, membrane bone periosteal cells can undergo chondrogenic differentiation. The precursor of chondrogenic cells in membrane bone is not clear: chondrocytes were proposed to arise from unipotential chondroprogenitor cells, bi- or multipotential progenitor cells, or differentiated osteogenic cells. There is experimental support for each, but studies on clonal and cell cultures provided more support for a common precursor of both chondro- and osteogenic cells. Moreover, in periostea, chondrogenesis probably arises from a differentiated cell type. Membrane bone formation in periostea may include a transient cell stage that is able to undergo both osteo- and chondrogenesis. Osteogenesis would be the normal pathway, but chondrogenesis can be evoked in certain microenvironments. It is not known whether microenvironmental factors trigger chondrogenesis through a universal molecular mechanism, nor is the molecule that triggers chondrogenesis known. Expression of neural cell adhesion molecule (NCAM) is down-regulated during commitment of periostal cells for secondary chondrogenesis, suggesting a possible regulatory role for NCAM in the alternative differentiation pathways of periosteal cells. PMID- 9406839 TI - Retrograde and anterograde labeling of cerebellar afferent projection by the injection of recombinant adenoviral vectors into the mouse cerebellar cortex. AB - Adenoviral vectors have recently been recognized as highly efficient systems for gene delivery into various tissues. We show that a reporter gene introduced into nerve terminals via an adenovirus can be used to label cell bodies retrogradely and then label the axons and nerve terminals of the infected neurons anterogradely in vivo. We injected a replication-defective recombinant adenovirus carrying the E. coli beta-galactosidase gene (lacZ) into the cerebellar cortex of the adult mouse. The first evidence of retrograde labeling was obtained at 2 days after the infection when neurons in the pontine nuclei and the reticulotegmental nucleus of the pons weakly expressed beta-galactosidase, and at 3 days post infection when neurons in all precerebellar nuclei, known to project to the cerebellar cortex, were strongly stained with X-gal in a Golgi-like manner. Anterograde transport of lacZ gene products was recognized at 3 days post infection; beta-galactosidase-positive axons arose from somata or dendrites of retrogradely labeled neurons, passed through the middle or inferior cerebellar peduncles, and entered the cerebellum. Anterogradely labeled mossy terminals were recognized on the injection side at 8 days post-infection, and on the contralateral side at 14 days post-infection. Beta-galactosidase expression persisted for up to two months, with a decrease in the total number of labeled cells over time. We could not find any signs of anterograde or retrograde transsynaptic labeling in the nuclei synaptically linked to the cerebellar cortex at any time point after injection up to 58 days post-infection. PMID- 9406840 TI - Apoptosis in the development of the temporomandibular joint. AB - Apoptosis has been shown to be involved in remodeling of organs during development, and derangement of the apoptotic process may result in temporomandibular joint (TMJ) dysfunction or congenital malformation. To investigate the relationship between the development of the TMJ and apoptosis, rat fetuses at 17.5-20.5 days of gestation (E17.5-20.5, vaginal plug=E0) and rats at postnatal days 1, 2, 3, 5, and 10 (P1, 2, 3, 5, and 10) were examined by light (LM) and transmission electron microscopy (TEM) and electrophoretic analysis of DNA fragmentation. At E17.5 and 18.5, a few layers of slender mesenchymal cells which eventually develop into the TMJ disk were observed, although TEM or electrophoresis did not reveal apoptotic cells at these stages. At E19.5 and 20.5, all structures of the TMJ except the lower joint cavity could be distinguished, but at these stages apoptotic cells were not observed. In P1 condyles, apoptotic cells were observed by TEM both at the subsurface of the condyle and in the region at which the lateral pterygoid muscle attaches to the condyle. These apoptotic cells showed irregular chromatin condensation, convolution of the cell membrane, and fragmentation and disintegration of the cytoplasm. Electrophoretic analysis of the P1 condyle further confirmed DNA fragmentation. Apoptosis was not observed in all specimens at the P1 stage. It was confirmed in 8 out of 20 animals (10 out of 27 joints) by TEM and/or electrophoretic analysis. The shape of the upper portion of the condyle flattened progressively from E20.5 to P2. At this stage, the lower joint cavity was developing, as observed by LM. These findings suggest that the morphological changes of the mandibular condyle effected by apoptosis, together with development of the lower joint cavity, play important roles in the postnatal functional adaptation to external stimuli such as mechanical strain. PMID- 9406841 TI - Postnatal development of interhemispheric asymmetry in the cytoarchitecture of human area 4. AB - The postnatal development of interhemispheric asymmetry was analyzed in the primary motor cortex (area 4) of 20 human brains with quantitative cytoarchitectonic techniques. The volume fraction of cortical tissue occupied by cell bodies (grey level index) was determined by automated image analysis. In children as well as in adults, the volume fraction of cell bodies averaged over all cortical layers was greater on the right than on the left. Thus, the space between cell bodies, i.e. the volume fraction of neuropil containing axons, dendrites and synapses, was greater in the left than in the right primary motor cortex. At the level of single layers, however, interhemispheric asymmetry of the neuropil volume fraction differed between age groups. The supragranular layers were significantly less asymmetrical in children than in adults, whereas the infragranular layers showed a similar degree of asymmetry in both age groups. Thus, the postnatal development of the architectonic asymmetry in the supra- and infragranular layers of area 4 follows the same sequence of maturation as found during neuronal migration, i.e. an inside-to-outside gradient. Comparing the layer-specific developmental pattern with available functional data, it was found that the structural maturation of interhemispheric asymmetry in the supragranular layers correlates with the development of hand preference. PMID- 9406842 TI - Expression of a low-molecular-weight (10 kDa) calcium binding protein in glial cells of the brain of the trout (Teleostei). AB - Calcium-binding proteins of the EF-hand family are widely distributed in the vertebrate central nervous system. In the present study of the trout brain, immunocytochemistry with a monoclonal antibody against chick gut calbindin-28k and a polyclonal antibody against bovine S100 protein specifically stained ependymocytes and radial glia cells with identical patterns. Western blot analysis of trout brain extracts with the antibodies to S100 and calbindin stained the same low-molecular-weight (10 kDa) protein band. In rat brain extracts, however, the monoclonal antibody to calbindin recognized a major protein band with molecular weight corresponding to that of calbindin-28k. This indicates that the trout protein is a new calcium-binding-like (calbindin-like) molecule that is immunologically related to both S100 and calbindin. Immunocytochemical studies of the trout brain using the antibodies to CaB and S100 showed that ependymocytes were stained in most ventricular regions, except in a few specialized ependymal areas of the ventral telencephalon, epithalamus, hypothalamus (including the paraventricular organ and saccus vasculosus) and brain stem. Immunocytochemistry also indicated the presence of calbindin-like protein in radial glia cells of several regions of the brain (thalamus, pretectal region, optic tectum, and rhombencephalon). Differences in immunoreactivity between neighbouring ependymal areas suggest that this protein may be a useful marker of different territories. All immunoreactive glial cells were nicotin adenin-dinucleotide-phosphate diaphorase-positive, although this enzymohistochemical reaction is not specific for these glial cells since it reveals oligodendrocytes and some neurons. Immunoreactivity appears at different developmental stages in the different brain regions, with a broadly caudorostral gradient, suggesting that the expression of this protein is developmentally regulated. Comparison of the distribution of the calbindin-like protein with that of glial acidic fibrillary protein indicates that calbindin-like immunocytochemistry is a specific technique for revealing radial glia and ependymocytes in the trout. PMID- 9406843 TI - Steroidogenic acute regulatory protein: the StAR still shines brightly. PMID- 9406844 TI - Steroid-independent activation of androgen receptor in androgen-independent prostate cancer: a possible role for the MAP kinase signal transduction pathway? PMID- 9406845 TI - Molecular characterization of neurotrophin expression and the corresponding tropomyosin receptor kinases (trks) in epithelial and stromal cells of the human prostate. AB - The prostate is one of the most abundant sources of nerve growth factor (NGF) outside of the nervous system. NGF is a member of the neurotrophin family of growth factors which in mammals also includes brain derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3) and neurotrophin-4/5 (NT-4/5). These neurotrophins can bind with high affinity to a family of tropomyosin receptor kinases (trks). These receptors are trkA, which binds NGF; trkB, which binds both BDNF and NT 4/5; and trkC, which binds NT-3. In order to characterize the molecular expression of the neurotrophins and their corresponding trk receptors in the prostate we performed Northern blot analysis for the neurotrophins and reverse transcription-polymerase chain reaction (RT-PCR) coupled with Southern blot analysis for the trk family of receptors on smooth muscle stromal cells from the prostate, the androgen responsive LNCaP prostate tumor cell line and the androgen refractory TSU-pr1 prostate tumor cell line. The results show that smooth muscle stromal cells expressed NGF, BDNF and trkC, whereas both epithelial cell lines expressed trkA, trkB and trkC to various degrees. NT-3 was not detected in either the smooth muscle stromal cells or in both epithelial cell lines. This suggests that the stromal cell derived NGF and BDNF may interact via paracrine mechanisms with trkA and trkB receptors, respectively, on the adjacent epithelial cells. Interestingly, the androgen responsive LNCaP cell line did not express any of the neurotrophins, whereas the androgen refractory TSU-pr1 cell line expressed NGF, BDNF and NT-4/5. This suggests that the autocrine expression of NGF, BDNF and NT 4/5 is up-regulated in prostate epithelial cells following their transformation to an androgen refractory pathology. Hence, the malignant transformation of prostate epithelial tumor cells may facilitate their escape from a paracrine dependence on stromal cell derived neurotrophins by the acquisition of the autocrine expression of neurotrophins. Since the pathology of malignant cell migration within the prostate is predominantly by direct extension around prostatic nerves the upregulation of autocrine neurotrophin expression within prostate epithelial tumor cells may be concomitant with transformation to a malignant phenotype capable of invasion along the perineural space and extracapsular metastasis to distant sites of tumor formation. PMID- 9406846 TI - L-thyroxine directly affects expression of thyroid hormone-sensitive genes: regulatory effect of RXRbeta. AB - L-thyroxine (T4) has been considered mainly a prohormone, the hormonal action of which is related to its conversion to 3,5,3'-triiodothyronine (T3) in peripheral tissues. In this study we investigated in transient transfection assays whether T4 might directly affect the expression of thyroid hormone (TH) sensitive genes. The reporter construct ME-TRE-TK-CAT or TSH-TRE-TK-CAT containing the nucleotide sequence of the TH response element (TRE) of either malic enzyme (ME) or TSHbeta genes, was transfected with either TH receptor (TR) alpha alone or in combination with retinoid X receptor (RXR) beta into NIH3T3 cells. Addition of 100 nM T4 to the culture medium in the presence of TRalpha increased the basal level of ME-TRE TK-CAT expression by 4.5-fold. T4 action was due to a direct interaction with TRalpha and not to its conversion to T3, since T4 effect persisted in the presence of 5'-deiodinase inhibitors (propylthiouracil, iopanoic acid) effectively preventing T3 generation, as assessed by the absence of T3 by HPLC in the cellular extracts of transfected cells. In a dose-response study half-maximal stimulation by T4 was achieved at a concentration of 100 nM, whereas 50% of maximal induction was produced by 1 nM T3 and 6 nM triiodothyroacetic acid (TRIAC). Coexpression of RXRbeta greatly enhanced the transcriptional activity of the ME-TRE-TK-CAT gene when either T3, T4 or TRIAC was added to the culture medium of NIH3T3 cells, but established a hormonal hierarchy in the reporter activation different than that observed in the presence of TRalpha alone (TRIAC > T3 > or = T4, instead of T3 > TRIAC > T4). T4 at a concentration of 100 nM could activate the TH/TR-dependent down-regulation mediated by the negative TSH-TRE, although at a lower level than that obtained with similar concentrations of T3 (35 and 55% inhibition, respectively). Our results demonstrate that, in addition to the action mediated through its monodeiodination to T3, T4 exerts a direct effect on genes that are either positively or negatively regulated by TH. Moreover, RXRbeta, forming heterodimers with TRs, appeared to exert a central role in modulating the sensitivity of TH-responsive genes to different iodothyronines. PMID- 9406847 TI - Ontogeny of 17beta-hydroxysteroid dehydrogenase type 2 mRNA expression in the developing mouse placenta and fetus. AB - 17beta-Hydroxysteroid dehydrogenase type 2 (17HSD type 2) catalyzes the inactivation of estradiol, testosterone and dihydrotestosterone into biologically less active 17-keto forms. Our recent Northern analysis indicated that the enzyme is expressed both in mouse placenta and fetus. The present data indicate that in the placenta the distribution of enzyme expression changes during pregnancy. In the choriovitelline placenta (day 8) 17HSD type 2 was expressed both in mural and polar giant cells. Later, on days 9-12.5, the mRNA was also detected in the junctional zone, and in late gestation (days 14.5-17.5), 17HSD type 2 mRNA was predominantly expressed only at the labyrinth region. In the fetus, 17HSD type 2 expression appears in the liver on day 11. At day 12 the expression was strongly increased in the liver, and at the same time moderate mRNA expression was also detected in the esophagus and intestine. In these tissues, high constitutive expression of 17HSD type 2 was then maintained throughout pregnancy. At later stages of development (days 15-16) the mRNA was, furthermore, detected in epithelial cells of the stomach, tongue, oropharynx and nasopharynx as well as in the kidney. We conclude that the expression pattern of 17HSD type 2 in the developing placenta and fetus suggests a role for the enzyme in maintaining a barrier to the transfer of active 17-hydroxy forms of sex steroids between the fetus and maternal circulation. PMID- 9406848 TI - C/EBP activates the human corticotropin-releasing hormone gene promoter. AB - The purpose of these studies was to identify whether transcription factors, associated with cytokine signalling, affected promoter activity of the corticotropin releasing hormone (CRH) gene. Fragments of a 3.6 kb sequence of the human CRH gene promoter were amplified by PCR and ligated upstream of a CAT reporter. These constructs were transfected into a variety of cell lines, either alone or together, with transcription factor expression vectors. Basal activity of a 3070 bp CRH promoter fragment was only seen in neuronal and lymphoblastoid cell lines. Promoter activity was increased by the transcription factors C/EBPbeta (NF-IL6) and more strongly, by C/EBPdelta (NF-IL6beta). Increased CRH promoter activity following phorbol ester treatment was inhibited by a dominant negative NF-IL6 mutant, showing that the effects of phorbol ester were principally mediated by C/EBP. Moreover, the inverse changes in the expression of CRH in the hypothalamus and spleens of arthritic rats were paralleled by similar inverse changes in NF-IL6beta expression in these organs. These data show that some transcription factors associated with cytokine signalling can also activate the CRH promoter. PMID- 9406849 TI - Ultra-structural characteristics of bovine granulosa cells associated with maintenance of oestradiol production in vitro. AB - We investigated whether the maintenance of oestradiol production by bovine granulosa cells (GC) in vitro was related to GC ultra-structure, and studied the effects of inclusion of serum as a cell attachment factor on oestradiol secretion, cell morphology and ultra-structure. Bovine granulosa cells from medium-sized follicles (4-8 mm diameter), in a serum-free (SF) culture system, maintained oestradiol production for 6 days, whereas oestradiol secretion by cells cultured in serum-coated (SC) wells declined rapidly with time, in culture. SF cells formed clumps consisting of two types of cells. Cells within clumps presented a phenotype similar to GC in vivo, being spherical, tightly joined by extensive gap junctions and interdigitated pseudopodia/microvilli, had abundant rough and smooth endoplasmic reticulum (ER) and mitochondria with trabecular cristae. In contrast, cells cultured in either SC wells or in the flattened base of cell clumps from SF cultures were enlarged, containing less rough ER, had fewer mitochondria (which tended to be round) and contained endosome-like structures, morphological characteristics suggestive of early luteinisation. PMID- 9406851 TI - Co-purification of a ribonuclease and human chorionic gonadotrophin beta-core protein from human urine and displacement of 125I-human luteinizing hormone from Candida albicans binding sites by ribonucleases. AB - An 18 kDa pregnancy urine protein preparation, purified to apparent electrophoretic homogeneity as judged by silver-staining of polyacrylamide gels, inhibited binding of 125I-hLH (human luteinizing hormone) to Candida albicans microsomes, reacted with monoclonal and polyclonal antibodies raised against human chorionic gonadotrophin (hCG) beta-core protein and exhibited ribonuclease (RNase) activity. Eleven of the 12 amino acids at the N-terminus of a protein in this preparation were identical to those of the N-terminus of human non-secretory ribonuclease. These results indicate co-purification of hCG beta-core with a RNase. An 18 kDa RNase was also purified from a commercial hCG preparation (Chorulon). However, no RNase activity was detected in a highly purified commercial preparation (Profasi). Three commercial RNase preparations displaced 125I-hLH from C. albicans binders at extremely low concentrations (< 0.001 microg/ml RNase) whereas only slight displacement of 125I-hLH from sheep luteal binding sites was observed with very high concentrations of the RNases (100 microg/ml RNase). The co-purification of hCG beta-core and RNase from pregnancy urine and the displacement of 125I-hLH from C. albicans binding sites by RNases may be related to the close relationship that has been identified between mammalian RNase inhibitors and the extracellular domain of gonadotrophin receptors. The presence of RNase in commercial preparations of gonadotrophins should be borne in mind during any investigations that involve impure preparations of these hormones. PMID- 9406850 TI - Sequence of the bovine HDL-receptor (SR-BI) cDNA and changes in receptor mRNA expression during granulosa cell luteinization in vivo and in vitro. AB - Steroidogenic activity in the mature corpus luteum of most mammals depends upon provision of cholesterol from the circulating lipoproteins. In cattle, as in many species, high-density lipoprotein (HDL) is the major lipoprotein involved. The recent identification of the scavenger receptor SR-BI as an HDL-receptor allows control of this process to be investigated more closely. In this study, we have sequenced the bovine SR-BI HDL-receptor and examined changes in expression of the receptor mRNA during corpus luteum development in vivo and granulosa cell luteinization in vitro. Sequencing of the bovine HDL-receptor showed that it codes for a protein of 509 amino acids with close identity to hamster, mouse, rat and human sequences. Examination of the tissue distribution of the HDL-receptor mRNA showed high levels in adrenal cortex and corpus luteum and lower levels in spleen and liver. Using a semi-quantitative, reverse transcription-polymerase chain reaction technique levels of HDL-receptor mRNA were measured in corpora lutea from cattle at known stages of the oestrus cycle and in bovine granulosa cells luteinized in culture. Levels of HDL-receptor mRNA were low in isolated bovine granulosa cells, but increased 7-fold during corpus luteum development in vivo and 5-fold during granulosa cell luteinization in culture. Results show that luteinization of granulosa cells is associated with an increase in HDL-receptor RNA levels which, along with changes in steroidogenic enzyme activity, is likely to explain the marked increase in steroidogenic capacity which occurs during corpus luteum formation. PMID- 9406852 TI - Pseudohypoaldosteronism: mutation found, problem solved? AB - The term 'pseudohypoaldosteronism' includes at least three distinct clinical syndromes, classified as type I, II and III, which differ in their clinical and biochemical findings but have in common the symptoms of mineralocorticoid resistance. The finding of a defect in the recently cloned epithelial sodium channel (ENaC) in a subgroup of familial pseudohypoaldosteronism type I has changed our understanding not only of the pathophysiology of these disorders but also the physiology of renal salt and water homeostasis. In this review the various clinical, biochemical and genetic findings in the different forms of pseudohypoaldosteronism will be discussed with the aim of identifying the underlying differences and similarities. The direction of further genetic investigations will depend at least in large part on further clinical classification of patients and families. PMID- 9406853 TI - Leukocytes modulate 11beta-hydroxysteroid dehydrogenase (11beta-HSD) activity in human granulosa-lutein cell cultures. AB - It is well established that there are interactions between the immune and reproductive systems. The ovary contains indigenous macrophages, as well as other classes of leukocytes in smaller numbers. Cytokines secreted by these cells have been shown to have the ability to regulate ovarian steroidogenesis. In the present study, the effect of leukocytes on 11beta-hydroxysteroid dehydrogenase (11beta-HSD) in human granulosa-lutein cells was examined. In addition, individual cytokines were also tested for their ability to regulate this enzyme. The follicular aspirates of patients undergoing IVF treatment were used as a source of granulosa cells. Cells isolated from these aspirates were found to contain between 15 and 60% leukocytes as assessed by flow cytometry (FACS). Leukocytes were removed from the sample preparations by the use of immunomagnetic beads coated with CD45 antibody, which recognises a surface antigen on all classes of leukocyte. Removal of leukocytes significantly decreased the 11beta HSD activity in the granulosa cells, assayed after 3 days of culture, from 7.3 (2 20) to 3.5 (1-10) pmol cortisone formed/50000 cells/4 h (medians and ranges, n = 15). Addition of IL-5 and IL-6 significantly increased the 11beta-HSD activity in granulosa cell cultures both in the presence and absence of leukocytes. Addition of IL-4 and IFN-gamma increased 11beta-HSD activity only in the leukocyte depleted granulosa cell cultures, whereas IL-2 had no effect on either of the cultures. The data suggests that leukocytes interact with the ovarian cells through cytokine secretion and/or cell-cell contact to increase the 11beta-HSD activity in human granulosa cells. PMID- 9406854 TI - Regulatory capacity of an androgen-specific enhancer of the mouse Slp gene in transgenic mice. AB - Different steroid hormone receptors can activate transcription from the same hormone response element (HRE) in vitro, but in vivo the effects of each hormone on gene activity are distinct. To determine sequences mediating androgen-specific response in a physiological setting, we placed the androgen-responsive mouse sex limited protein gene (Slp) enhancer before a tkCAT reporter in transgenic mice. The enhancer contains a consensus HRE plus accessory factor binding sites that act in concert to direct transcription in response to androgen. A 160 bp fragment, C'delta2, is responsive to several steroids in transfection; in transgenic mice, this enhancer was active in several tissues of male and female mice, in four of six transgenic lines. In striking contrast, C'delta9, a 120 bp sub-fragment of C'delta2 that responds only to androgen in transfection, showed activity in testes, prostate and kidney, where it was strongly androgen-inducible in females. However, expression was obtained in only one transgenic line. Multimerization of the C'delta9 enhancer conferred expression in prostate, but again in only one line. The greater penetrance of C'delta2 expression was not driven by glucocorticoids, as adrenalectomy had little effect, but may be dependent on the NF-kappaB-like element absent from the C'delta9 fragment. That two transgenic lines showed expression in androgen target sites driven by enhancers that are androgen-specific in vitro suggested that activation of this enhancer, when it could occur, was in response to androgen. The dramatically different behavior of the two related enhancer sequences underscores the importance of chromosomal context to the activity and specificity of regulatory elements. PMID- 9406855 TI - The full agonistic effect of recombinant 20 kDa human growth hormone (hGH) on CHO cells stably transfected with hGH receptor cDNA. AB - The agonistic effect of the recombinant 20 kilodalton human GH (20K-hGH) with authentic primary structure was studied using Chinese hamster ovary (CHO) cells stably transfected with hGH receptor (hGHR) cDNA and was compared with that of 22K-hGH. The binding affinities (dissociation constants) of 20K- and 22K-hGH were identical (0.41 +/- 0.11 nM and 0.41 +/- 0.04 nM, respectively). In addition, the two hGHs possessed the same potencies in activating the rat serine protease inhibitor (Spi) 2.1 gene promoter. 20K-hGH was similarly internalized as 22K-hGH but its internalization rate was a little slower than that of 22K-hGH. We also found that proliferation of CHO-hGHR cells stimulated by serum was remarkably inhibited by both hGHs to the same degree. In conclusion, both hGH isoforms exhibited the same binding affinities for hGHR and were potent enough to induce some hGHR-mediated cellular events. These suggest that 20K-hGH exerts a full agonistic activity for hGHR. PMID- 9406856 TI - Temporal expression of urokinase type plasminogen activator, tissue type plasminogen activator, plasminogen activator inhibitor type 1 in rhesus monkey corpus luteum during the luteal maintenance and regression. AB - Proteolytic activity generated by the plasminogen activator (PA) system has been associated with many biological processes. Using a pregnant mare serum gonadotropin (PMSG)/human chorionic gonadotropin (hCG)-induced rhesus monkey corpus luteum (CL) model, we have studied how urokinase-type plasminogen activator (uPA), tissue-type plasminogen activator (tPA), and plasminogen activator inhibitor type 1 (PAI-1), are temporally expressed in CL of rhesus monkey at the luteotropic and luteolytic periods. Slot blot analysis and in situ hybridization were performed to analyze the expression and distribution of uPA and PAI-1 messenger RNA (mRNA). Fibrin overlay was used to detect uPA and tPA activities. We found that uPA is the dominating PA in luteotropic CL in the monkey. Abundant expression of PAI-1 mRNA was detected. The highest expression of uPA and PAI-1 mRNA was observed at the luteotropic period, while their expression decreased approximately 50% at early luteal regression defined by considerably decreased serum progesterone levels, and remained at very low levels at the late stage of luteal regression. We also observed an increased tPA activity at the time of luteal regression. Moreover, the exogenous tPA could inhibit the progesterone production in cultured luteal cells from 13-day-old monkey CL. We also used LH receptor mRNA expression as a mark for the luteal phases. A highly expressed, evenly distributed LH receptor mRNA was detected in CL during the luteotropic phase, while its expression decreased at day 13 coinciding with the reduction of progesterone production. We conclude that proteolysis mediated by uPA and regulated by PAI-1 may play a role in the luteal maintenance, while tPA may participate in the luteal regression in the rhesus monkey. PMID- 9406857 TI - Glucocorticoids regulate L-fucose glycoconjugates in rat pancreatic zymogen granules. AB - Lectin-binding studies were performed on rat pancreatic zymogen granules to investigate the influence of glucocorticoid levels on saccharide membrane composition. The following animal groups were used: (1) control rats; (2) rats treated with hydrocortisone (1, 10 and 25 mg/kg/day) for 1, 3 and 8 days; (3) postadrenalectomized rats at days +1, +3 and +8; and (4) adrenalectomized rats receiving hydrocortisone therapy (10 mg/kg/day) for 8 days. By flow cytometry, fluoresceinated (FITC) lectins were used to measure the amount of Concanavalin A (Con A) (specific for D-mannose), wheat germ agglutinin (WGA) (specific for N acetyl-D-glucosamine) and sialic acids and Tetragonolobus purpureus (TP) (specific for L-fucose) bound to individual zymogen granules from two subpopulations, Z1 and Z2, identified on the basis of their forward and side scatter properties. The molar ratio of the different FITC-lectins revealed significant differences in the glycoconjugate composition of Z1 and Z2 granules, the Z1 granules showing higher ratios of N-acetyl-D-glucosamine:L-fucose and N acetyl-D-glucosamine:D-mannose, both in control, adrenalectomized and hydrocortisone-treated rats. It was also observed that N-acetyl-D-glucosamine and/or sialic acids were more abundant than L-fucose and D-mannose in the zymogen granule membrane. Z1 and Z2 granules had different glycosylation patterns. Neither adrenalectomy nor hydrocortisone treatments varied the Con A binding to zymogen granules. An increase in WGA binding was only induced by administration of very high doses of hydrocortisone (25 mg/kg/day) for 8 days, an effect not directly related to glucocorticoids. In contrast, a correlation between the FITC TP labelling and glucocorticoid levels can be established, so that, in a time dose dependent way, an increase was observed in zymogen granules of rats treated with hydrocortisone while a decreased TP binding was found in adrenalectomized rats-an effect which was reversed with hydrocortisone therapy. Therefore, glucocorticoids exert a direct influence on the saccharide composition of rat pancreatic zymogen granules, regulating the amount of L-fucose glycoconjugates, with Z2 granules more sensitive than Z1 ones. PMID- 9406858 TI - Localisation and regulation of 17beta-hydroxysteroid dehydrogenase type 3 mRNA during development in the mouse testis. AB - The final step in the biosynthesis of testosterone is the reduction of androstenedione to testosterone catalysed by the enzyme 17beta-hydroxysteroid dehydrogenase (17betaHSD). Five isoforms of the enzyme have been identified in the mouse and the type 3 isoform has been shown to be the predominant reductive form present in the adult human and mouse testis. In this study the regulation of 17betaHSD type 3 isoform mRNA levels and the cellular localisation of the enzyme mRNA have been studied in the mouse testis. To examine regulation of 17betaHSD type 3 mRNA expression in the testis, mRNA levels were measured during development in normal mice and in mice lacking circulating gonadotrophins (hpg) or functional androgen receptors (Tfm). In these mutants testicular descent does not occur at the normal time (25 days) and control animals were, therefore, rendered cryptorchid at 19 days. In neonatal mice, it has been shown a peak of type 3 expression occurs around day 5 and this was found to be normal in all groups in the current study. In normal animals there was a marked increase in type 3 isoform expression between 25 and 30 days and this continued into adulthood. In cryptorchid animals the increase in type 3 mRNA levels after 25 days was less marked than in untreated controls and by 90 days was about 15% of normal animals. In Tfm mice, levels of 17betaHSD type 3 mRNA failed to show any increase around puberty (25 days) and in adult Tfm mice, levels were less than 1% of cryptorchid controls. In hpg mice, levels of type 3 mRNA increased slowly after puberty and were about 30% of cryptorchid controls by 90 days. Studies using in situ hybridisation showed that the type 3 isoform was expressed only in the interstitial tissue of the adult normal mouse testis. No specific hybridisation could be determined in adult hpg or Tfm testes. Results show that 17betaHSD type 3 is an interstitial enzyme in the testis and is, probably, localised in the Leydig cells. During neonatal development expression of 17betaHSD type 3 is independent of gonadotrophin action while the increase in type 3 expression at puberty is primarily dependent upon androgen action although testicular descent and gonadotrophins are also required. PMID- 9406859 TI - Ginsenoside-Rg1, one of the major active molecules from Panax ginseng, is a functional ligand of glucocorticoid receptor. AB - We have examined the possibility that a component of Panax ginseng, ginsenoside Rg1 (G-Rg1), acts by binding to the glucocorticoid receptor (GR). G-Rg1 competed for [3H]dexamethasone (Dex) binding to GR with a specific affinity of 1-10 microM and activated a glucocorticoid responsive element-containing luciferase reporter gene. The dose-dependence patterns of G-Rg1 and Dex for these two effects were nearly identical, although two to three orders of magnitude higher concentration of G-Rg1 than that of Dex was required for the same magnitude of response. At the cellular level, the growth of FT02B cells was suppressed by G-Rg1 as well as by Dex, each of whose effects were abolished by RU486. These results demonstrate that G-Rg1 is a functional ligand of GR. PMID- 9406860 TI - Mammary Stat5 abundance and activity are not altered with lactation state in cows. AB - Stat5 is a key intracellular mediator of prolactin signalling and can activate transcription of milk proteins in response to prolactin. Therefore, in animals such as mice where lactation is dependent on prolactin, Stat5 is likely to play an important role in establishing or maintaining lactation in the mammary gland. However, little is known about its role in lactation in the dairy cow. In order to address this, the levels of Stat5a and Stat5b protein, mRNA and Stat5 DNA binding activity were measured in mammary tissue from mice and cows at different lactational states. In the cow, Stat5a and Stat5b protein and mRNA levels, as well as Stat5 DNA-binding activity were unaltered between pregnancy and established lactation. In contrast, in the mouse Stat5a and Stat5b protein, as well as Stat5 DNA-binding activity were clearly increased during lactation whereas Stat5a and Stat5b mRNA levels were highest during pregnancy as has been previously described. In both species only a minority of the epithelial cell nuclei were Stat5 positive during established lactation. These results suggest that there are significant differences in the biological role of Stat5 in controlling lactation between ruminants and rodents. PMID- 9406861 TI - Glycemia-lowering effect of cobalt chloride in the diabetic rat: increased GLUT1 mRNA expression. AB - We have recently shown that expression of the GLUT1 glucose transporter isoform is augmented in cells exposed to cobalt chloride [Co(II)], an agent that stimulates the expression of hypoxia-responsive genes (Behrooz, A., Ismail-Beigi, F., 1997. J. Biol. Chem. 272, 5555-5562.). Here, we examine the effect of Co(II) on glycemia and tissue GLUT1 mRNA content of normal and diabetic rats. The addition of 2 mM Co(II) in the drinking water reduced the glycemia of streptozotocin-induced diabetic rats by day 3 from 32.3 +/- 2.1 to 21.0 +/- 1.9 mM (non-fasting). Co(II) resulted in no change in serum insulin levels of normal or diabetic rats. Treatment with 4 mM Co(II) was more effective than 2 mM Co(II) in reducing the glycemia of diabetic rats, while 6 mM Co(II) was associated with severe toxicity. GLUT1 mRNA content increased significantly in ventricular myocardium, renal cortex, skeletal muscle, cerebrum and liver of normal and diabetic rats treated with 2 mM cobalt chloride (ranging from 1.3- to 2.9-fold in the different tissues). It is concluded that: (1) treatment with Co(II) decreases the glycemia of diabetic rats, and (2) the glycemia-lowering effect of Co(II) is associated with, and may be mediated by, enhanced expression of GLUT1 mRNA. PMID- 9406862 TI - Repression participates in mammary tissue-specific activation of the caprine beta lactoglobulin promoter. AB - Activation of the beta-lactoglobulin (BLG) gene promoter is restricted to overtly differentiated mammary tissue. To understand the mechanism underlying such tissue specificity, activity of the caprine BLG promoter was analyzed comparatively in cultured mammary HC11 cells and non-mammary HeLa and CV-1 cells. The BLG promoter flanked by the 5'-regulatory sequence below -205 was strongly activated in the cells, regardless of the cell type. In non-mammary HeLa and CV-1 cells, this activation was repressed completely by the upper regulatory sequence. Weak repression was also observed in mammary HC11 cells kept non-confluent. As the mammary HC11 cells grew confluent and maintained the stabilized state, however, repression by the upper regulatory sequence was switched to activation. The repressive upstream flanking sequence was strongly recognized by the binding factors in non-mammary HeLa and CV-1 cells in an in vitro binding assay. Binding intensity and competition strength of the upstream regulatory regions were in a close correlation to their transcriptional repression activities in the cultured cells. The results suggest that the restricted activation of the caprine BLG promoter in differentiated mammary tissue is guaranteed by repression in non mammary and undifferentiated mammary cells. PMID- 9406863 TI - Mitogen-activated protein kinase kinase inhibition decreases growth hormone stimulated transcription mediated by STAT5. AB - We have investigated the possible involvement of the MAPK pathway in the growth hormone(GH)-induced activation of one of the members of signal transducers and activators of transcription, STAT5, by using the MAPK kinase (MEK) inhibitor PD98059. PD98059 treatment of Chinese hamster ovarian cells, stably transfected with the GH receptor (CHOA cells), abolished the GH-induced MAPK activity. PD98059 decreased the amount of GH-induced STAT5 in nuclear extract with DNA binding capacity. Furthermore, GH dependent transcription of a STAT5 regulated reporter gene was inhibited by PD98059. The MEK inhibitor did not reduce GH stimulated nuclear translocation of STAT5. We also investigated if PD98059 differentially influences the activation of the two STAT5 homologs, STAT5a and STAT5b, which differ mainly at the C-terminal end, one of the differences being the presence of a possible MAPK phosphorylation site in STAT5a. Expression plasmids for these transcription factors were transfected into CHOA cells together with a reporter gene. GH-stimulated fold induction of transcription was reduced by PD98059 in STAT5a but not in STAT5b overexpressing cells. A MAPK phosphorylation site-mutated version of STAT5a was also transfected into CHOA cells. GH-stimulated fold induction of cotransfected reporter gene was not reduced by PD98059 in cells overexpressing mutant STAT5a. The above data show that the MAPK pathway is required for the full activation of one of the STAT5 isoforms (STAT5a). PMID- 9406865 TI - D2 receptor-mediated inhibition of GABA release by endogenous dopamine in the rat globus pallidus. AB - Attempting to better understand the role of the dopaminergic innervation in the rat globus pallidus, we examined here whether or not endogenous dopamine modulates the release of [3H]GABA in superfused pallidal slices. The superfusion medium contained elevated (15 mM) potassium. The release of endogenous dopamine was induced by the dopamine releaser drug, methamphetamine. Methamphetamine (100 microM) inhibited by 46% the release of [3H]GABA. Methamphetamine inhibition was completely blocked by reserpinization of the rats. It was also completely blocked by the D2 dopamine receptor antagonist sulpiride (10 microM). Sulpiride alone caused a 105% increase in GABA release. The increase was not observed in slices from reserpinized rats. Quinpirole (10 microM), a D2 dopamine receptor agonist, inhibited (43%) [3H]GABA release. The results suggest that endogenous dopamine exerts an inhibitory effect on GABA release in the rat globus pallidus. The effect is mediated by D2 receptors presumably located on striatopallidal axon terminals. PMID- 9406864 TI - FGF-2 antisense RNA encodes a nuclear protein with MutT-like antimutator activity. AB - Bidirectional transcription of the basic fibroblast growth factor (FGF-2) gene gives rise to multiple polyadenylated sense mRNAs and a unique 1.5 kb antisense transcript (FGF-AS) which is complementary to the 3'-untranslated region of the FGF-2 mRNA. The rat FGF-AS cDNA encodes a novel 35 kDa nuclear protein (GFG) with homology to the MutT family of antimutator NTPases. Antibodies against the deduced amino acid sequence of GFG detected intense immunoreactivity in the nuclei of adult rat hepatocytes. Subcellular fractionation and Western blotting confirmed the presence of a 35 kDa immunoreactive protein in the nuclear fraction and, to a lesser extent, in the mitochondrial fractions of rat liver homogenates. Recombinant GFG suppressed the spontaneous mutation rate of MutT-deficient E. coli in a complementation assay. In-frame deletion of the 53 amino acids encompassing the MutT domain eliminated this activity, confirming the catalytic function of this region in the FGF antisense gene product. These findings demonstrate for the first time that the FGF-AS transcript encodes a functional nuclear protein with MutT-related enzymatic activity. PMID- 9406866 TI - Direct measurement of the chloride concentration in newt olfactory receptors with the fluorescent probe. AB - Chloride (Cl-) current in the olfactory cell has been proposed to be excitatory and amplifying the receptor current. As an intracellular concentration of Cl- ([Cl-]i) is critically important for the proposed function, we tried to measure [Cl-]i of the isolated newt olfactory receptors using chloride-sensitive fluorescent dye, N-(6-methoxyquinolyl)-acetoethyl ester (MQAE). We found that the average of Cl- concentration through the cell is about 40 mM. This value is fairly lower than 120 mM that was suggested from reversal potential of tail component of the olfactory response in the low Cl- bath solution. Because the reversal potential for the present [Cl-]i is above the resting potential, opening of the Cl- channels may serve as a booster for the depolarizing odor-response. PMID- 9406867 TI - Molecular identification of a dopamine D1b receptor in bovine retinal pigment epithelium. AB - The rhythmic daytime inhibition of phagocytosis of shed photoreceptor outer segments (OS) by the retinal pigment epithelium (RPE) is related to increased cAMP in RPE cells. Dopamine (DA), the light-adaptive signal of the retinal oscillator can activate adenylyl cyclase through its D1-like receptors. It reduces OS phagocytosis by cultured bovine RPE, but a DA receptor was not demonstrated. Using primers selected from alignment of D1-like receptor genes already cloned, we have amplified by PCR two sequences in bovine genomic DNA. Phylogenetic analysis demonstrated that they correspond to D1a and D1b receptors. These receptors were then searched for using the reverse transcription-polymerase chain reaction (RT-PCR) in cultured bovine RPE. Only the D1b receptor subtype was demonstrated. It could mediate the DA-induced inhibition of phagocytosis. PMID- 9406868 TI - Lipid mediators modulate NMDA receptor currents in a Xenopus oocyte expression system. AB - We determined the modulatory effects of various lipid mediators on mouse N-methyl D-aspartate (NMDA) receptor currents in the Xenopus oocyte expression system. Arachidonic acid, but not oleic acid potentiated NMDA receptor activity. The epsilon1/zeta1 heterodimer of the NMDA receptor was more sensitive to arachidonic acid than was the epsilon2/zeta1 heterodimer. Platelet-activating factor (PAF) and lysophosphatidic acid (LPA) both activated the NMDA currents, and the effects were more evident in the epsilon2/zeta1 heterodimer than in epsilon1/zeta1. These activations were abolished by treatment with protein kinase inhibitors, suggesting a possible phosphorylation of the receptor. Thus, lipid mediators do have modulatory effects on NMDA receptor currents, the potentiating effects of which differ depending on subtype of the NMDA receptor. PMID- 9406869 TI - Alteration of transcription factors NF-kappaB and STAT1 in Alzheimer's disease brains. AB - Recent studies suggest that inflammatory activation occurs in the brains of patients with Alzheimer's disease (AD). Several transcription factors such as nuclear factor-kappaB (NF-kappaB) and signal transducer and activator of transcription-1 (STAT1) may be activated in glial cells by a number of cytokines and then translocated from the cytosol to the nucleus. We assessed NF-kappaB and STAT1 in temporal cortex from normal and AD brains using specific antibodies. NF kappaB p65 in the particulate fraction and STAT1alpha in both the particulate and cytosolic fractions were more abundant in AD cases than in controls. These findings suggest that the increased NF-kappaB and STAT1alpha in cell nuclei may be involved in inflammatory activation in AD brains. PMID- 9406870 TI - The mismatch negativity component reveals the sensory memory during REM sleep in humans. AB - Auditory evoked potentials (AEPs) were recorded during presentation of stimuli of 1000 Hz (standard) and 2000 Hz (deviant) in trains of 10 tone bursts (one deviant per train) in the wake and rapid eye movement (REM) sleep states. The constant inter-stimulus interval (ISI) was 600 ms and the trains were separated by 3 s of silence. The deviant tone occurring at the train start elicited a mismatch negativity component (MMN) in both arousal states, displaying a peak latency between 100 and 150 ms post-stimulation at fronto-central areas. These results suggest the existence of an auditory memory trace (sensory memory) surviving for at least 3 s during REM sleep. PMID- 9406871 TI - Novelty-induced locomoter activity in Long-Evans rats pre- and post-chronic 'binge'-pattern cocaine treatment. AB - Incremental locomotor activity observed in behaviorally sensitized rats is associated with the activation of the mesocorticolimbic dopaminergic system and the hypothalamic-pituitary-adrenal (HPA) axis. To determine whether individual locomotor differences are altered in the behaviorally sensitized state, Long Evans rats were placed in a novel environment and locomotor activity was recorded for 2 h. Animals, thereby, were evenly divided into two activity groups: lower- (LR) and higher- (HR) responders (LR, 367 +/- 38 cumulative beam breaks; HR, 797 +/- 43; P < 0.01). Subsequently, rats were randomly assigned to saline or chronic 'binge'-pattern (CBP) cocaine (15 mg/kg i.p., three injections/day for 14 days) treatment groups. One hour after the last injection, rats were sacrificed and trunk blood was collected for plasma corticosterone (CORT) determination. CORT levels were higher in cocaine versus saline treated animals (P < 0.01). CBP cocaine treated rats had higher locomotor activity compared to saline treated animals (P < 0.05). Moreover, rats less vulnerable to psychostimulant self administration (LRs) appeared to have locomotor behavior resembling that of the more vulnerable phenotype (HRs) after CBP cocaine. These findings suggest that behavioral sensitization, as a result of CBP cocaine treatment, changes novelty stress induced behavior which may reflect altered individual vulnerability to drugs of abuse. PMID- 9406872 TI - Evidence that protein kinase C activation is involved in the excitatory and facilitatory effects of bradykinin on canine visceral nociceptors in vitro. AB - The involvement of protein kinase (PK) C activation in the effects of bradykinin (BK) on peripheral nociceptors, polymodal receptors, was examined using canine testis-spermatic nerve preparations in vitro. Phorbol 12,13-dibutyrate 0.1 microM, which activates PKC, suppressed the BK-induced excitation when applied for 3-5 min prior to BK application, but facilitated it when applied simultaneously with BK. Neither effect was induced by an inactive phorbol ester, 4alpha-phorbol 12, 13-didecanoate, demonstrating that both effects were mediated through the activation of PKC. In addition, staurosporine 1 microM, a PK inhibitor, suppressed both BK-induced excitation and facilitation of the heat response of testicular polymodal receptors without influencing on-going activities and the heat response itself. These results suggest that PKC activation is involved in the excitatory and facilitatory effects of BK on peripheral nociceptors. PMID- 9406873 TI - Activin facilitates neuronal development in the rat amygdala. AB - The amygdala is one of the richest sources for activin receptor in the central nervous system (CNS) but the function of activin in the amygdala is unknown. An in vitro culture system was developed to study the effect of recombinant human activin-A on neuronal growth. Activin-A (1000 pM) was added continuously from day 1 in vitro and the medium changed every 2 days. Continuous visual assessment revealed that control neurones started to atrophy within 2 days of incubation in serum free N2-MEM. After 6 days in culture, cells were fixed and stained for growth-associated protein (GAP-43), a membrane-bound phosphoprotein involved in axonal elongation and synaptogenesis. Activin-A reduced the number of atrophying neurones and stimulated neuritic growth. The results presented here indicate a possible neurotrophic role for activin-A in the neonatal CNS. PMID- 9406874 TI - Identification of the glial cell types containing carnosine-related peptides in the rat brain. AB - The cellular localization of carnosine-like immunoreactivity was investigated in the adult rat forebrain and in glial cell cultures obtained from newborn rat brain. Using double staining methods, we showed that in vivo carnosine-like immunoreactivity was occurring in a large number of both glial fibrillary acidic protein (GFAP)-positive astrocytes and 2'3'-cyclic nucleotide 3' phosphodiesterase (CNP)-positive oligodendrocytes. In vitro, the carnosine immunoreactive staining was restricted to a subpopulation of completely differentiated oligodendrocytes, whereas no reaction was detected in immature oligodendrocytes and in astrocytes. These observations could have profound physiopathological implications considering the role suggested for carnosine and related peptides as endogenous antioxidants, free radical scavengers and anti glycating agents of the central nervous system (CNS). PMID- 9406875 TI - 5-HT1A receptor mRNA expressions differ in the embryonic spinal cord of male and female rats. AB - During critical periods of development, the effects of testosterone (T) on promoting androgenization of the central nervous system (CNS) are reflected not only by behavior, morphology, and hormone secretion but also by gene expression. The mechanisms involved in sexual differentiation of the CNS, however, remain incompletely defined. The current set of experiments examined with in situ hybridization the dimorphism in 5-HT1A receptor mRNA expression in the embryonic rat spinal cord and the possible role of T in the dimorphism. We found sex related differences in expression of 5-HT1A mRNA in the spinal cord, which were altered by a single injection of T. The results suggest that this gonadal steroid is responsible for the sexual dimorphism in 5-HT1A mRNA expression occurring during the critical period. PMID- 9406876 TI - A metalloprotease-inhibitor reduces pain associated behavior in mice with experimental neuropathy. AB - Tumor necrosis factor-alpha (TNF) is involved in the generation of inflammatory and neuropathic pain. The synthetic hydroxamic acid based metalloprotease inhibitor TAPI blocks cleavage of cell surface TNF and thus reduces levels of the mature 17-kDa TNF polypeptide in activated macrophages and T-cells. We have previously shown that pharmacologic inhibition of TNF production reduces pain related behaviors in mice with chronic constriction injury (CCI). Here we investigated whether blockage of TNF shedding by administration of TAPI would diminish hyperalgesia in animals with partial nerve injury. We injected 0.5 mg of the inhibitor epineurially once daily to mice with CCI for 7 days. The animals were tested for withdrawal thresholds to heat to test for thermal hyperalgesia and to von Frey hairs to assess mechanical allodynia. Mice with CCI developed thermal hyperalgesia and mechanical allodynia by day 3 after the injury. In mice treated with TAPI, a reduction of thermal hyperalgesia and mechanical allodynia of up to 50% occurred. Endoneurial TNF-immunoreactivity was reduced, but not immunoreactivity for IL-1alpha or IL-1beta. The numbers of degenerating axons and endoneurial macrophages were not affected by the treatment as compared to controls. We conclude that the metalloprotease inhibitor TAPI specifically reduces endoneurial TNF-levels after nerve injury and thereby may diminish neuropathic pain in the CCI-model. PMID- 9406877 TI - Migration activity of microglia and macrophages into rat brain. AB - We examined the entry of intra-arterially injected microglia and macrophages into the brain using a rat muscle graft model to compare their respective abilities to invade the brain parenchyma. Isolated microglia without any activation treatment entered into the brain with or without the muscle graft, while macrophages activated by phorbol 12-myristate-13-acetate (PMA) entered the brain only in the presence of the muscle graft. These results suggest that microglia have a higher affinity for the brain than macrophages. PMID- 9406878 TI - A search for primitive Purkinje cells: zebrin II expression in sea lampreys (Petromyzon marinus). AB - Zebrin II/aldolase C is a 36 kDa polypeptide expressed by Purkinje cells in the cerebellum of elasmobranchs, teleosts, birds, and mammals, and by octavolateralis pyramidal cells in developing teleosts. To better understand the evolution of these two systems we determined if zebrin II is expressed (1) in previously described primitive Purkinje cells, and (2) in octavolateralis pyramidal cells of sea lampreys (Petromyzon marinus). Ammocete and adult stages were reacted with mab anti-zebrin II. In ammocetes the large pyramidal cells of the anterior octavomotor nucleus (AON) were mab anti-zebrin II immunoreactive, but immunoreactivity was not detected in the cerebellar plate. In adults there was no immunoreactivity in any portion of the brain, including the cerebellar plate and the AON. The data indicate that zebrin II immunoreactivity may prove valuable in studying the development of the octavolateralis system across vertebrates. Three explanations are proposed to account for the absence of zebrin II+ Purkinje cells: aldolase C is expressed in Purkinje cells but the zebrin II epitope has not yet evolved; the zebrin II epitope was present in ancestral lampreys but has since been lost; or sea lampreys do not have Purkinje cells. The evolutionary implications of these results are briefly reviewed. PMID- 9406879 TI - Estrogen blocks neurotoxic effects of beta-amyloid (1-42) and induces neurite extension on B103 cells. AB - Clinical studies have shown that estrogen replacement therapy is associated with reduced risk of Alzheimer's disease (AD). We tested whether or not estrogen blocks neurotoxic effects of beta-amyloid (1-42) (A beta1-42) on cultured B103 cells. A beta1-42 (1 microM) induced typical necrotic cell death, as revealed by light and electron microscopic examinations. Co-administration of estrogen not only blocked A beta1-42 toxicity to a large degree, but also enhanced neurite extension. Pretreatment with estrogen was even more effective in blocking A beta1 42 toxicity. When added 18 h after the beginning of A beta1-42 treatment, estrogen was still effective in halting the progress of cell death and enhancing neurite extension. The protection against A beta1-42-induced neuronal death by estrogen was unlikely due to a blockade of lipid peroxidation injury, since estrogen completely failed to attenuate ferrous chloride-induced cell death. These results demonstrate that estrogen blocks A beta1-42-induced neurotoxicity and enhances neurite extension on B103 cells, both of which may well be underlying mechanisms of beneficial effects of estrogen in AD. PMID- 9406880 TI - Human brain potentials to reading syntactic errors in sentences of different complexity. AB - In order to determine if an event-related brain potential (ERP) effect described for syntactic violations (P600/SPS) varies with the amount of reprocessing entailed by a violation, number incongruencies were presented either within simple declarative or within subordinate clauses. ERPs were recorded while 12 German subjects read the stimulus materials presented word by word on a video monitor. The ERPs showed a P600/SPS effect for all sentence types, which was smallest in amplitude and earliest in latency for simple declarative sentences. This effect therefore qualifies as a metric for the amount and timing of syntactic reprocessing entailed by a syntactic error. In addition, a late frontal negativity (1000-1400 ms range) was found for the simple declarative sentences. PMID- 9406881 TI - Human equilibrium on unstable support: the importance of feet-support interaction. AB - Healthy humans maintained equilibrium on rocking supports (seesaw) of different curvatures and heights. We recorded platform tilt, horizontal displacements of the upper body, ankle joint angle and activity of ankle joint muscles. Subjects maintained balance by making seesaw rotations placing the support under the body's centre-of-gravity. Forward displacement was balanced by compensatory plantariflexion: thus the relation between muscle activity and ankle joint angle differed from that on a rigid floor. Mechanical analysis of stability showed that standing on low seesaws requires ankle torque increase during forward body shift (as on a rigid floor) and torque decrease on high seesaws (when the seesaw height exceeded its radius). In the latter case, balancing was impossible with eyes closed. The results suggest that directionally specific torque changes in response to centre-of-gravity shifts provide important information for maintenance of orthograde posture. PMID- 9406882 TI - Calcitonin gene-related peptide- and substance P-like immunoreactive fibers in the spermatic nerve and testis of the dog. AB - In order to determine if calcitonin gene-related peptide (CGRP) and substance P (SP) coexist in peripheral spermatic nerve fibers, we carried out a double staining immunofluorescence study using confocal microscopy and fluorescence microscopy. CGRP- and SP-like immunoreactivity (LI) coexisted in the spermatic nerve trunk and in the single fibers running along the surface of the testis. The great majority of the SP-containing fibers also held CGRP-LI, although some fibers contained CGRP-LI without SP-LI. These observations are consistent with previous observations on testicular dorsal root ganglion neurons. Additionally, we carried out an immunogold silver staining for CGRP and found CGRP-containing nerve bundles, single nerve fibers and their nerve terminals. Some CGRP containing nerve terminals were located very superficially in the tunica albuginea (<5 microm from the surface). PMID- 9406883 TI - Characterization of ligand binding properties of the 5-HT1D receptors cloned from chimpanzee, gorilla and rhesus monkey in comparison with those from the human and guinea pig receptors. AB - The 5-HT1D receptor is a potential target of anti-migraine drugs, and here its genes were cloned from chimpanzee, gorilla and rhesus monkey, via polymerase chain reactions with their genomic DNAs and the primers designed from the 5' and 3' untranslated regions of the human receptor. Direct sequencing of the polymerase chain reaction (PCR) products revealed high degrees of identity between their deduced amino acid sequences (the chimpanzee, gorilla and rhesus monkey) and that of human, differing by two, four and 11 residues, respectively. The binding properties of the receptors, as expressed in human embryonic kidney 293 cells, were compared to those obtained with the human and guinea pig receptors, the latter differing by 33 residues from the human receptor. Standard serotonergic ligands including several indoles, ergots and methiothepin bound all the cloned primate and guinea pig receptors with comparable, low nanomolar affinities, leading to high correlation coefficients among their Ki values. R(+) 8-Hydroxydipropylaminotetralin, on the other hand, bound the human receptor with the affinity higher than those for the primates and guinea pig receptors. This indicates that certain chemical templates may differentiate the molecular divergences among the 5-HT1D receptors of various animal species, and the use of the non-human primates will be beneficial for pharmacological characterizations, more relevant to the human receptor, of future novel ligands for the 5-HT1D receptor, which are potential anti-migraine drugs. PMID- 9406884 TI - Time course of human 40 Hz EEG activity accompanying P3 responses in an auditory oddball paradigm. AB - In order to quantify the time course of auditory P3-related gamma activity, root mean square (RMS) values were calculated from band-filtered (30-45 Hz) target and non-target responses in an auditory oddball experiment. Evoked (phase locked) gamma activity was evaluated from the time domain averages, whereas induced (not necessarily phase locked) activity was analyzed on the basis of single trials. Gamma RMS values were integrated across different time windows, namely the prestimulus, N50/P50, N100, pre P3, P3 and post P3 window. The single trial P3 window hereby was defined by a maximum amplitude criterion. In accordance with other studies, we found a pronounced increase of evoked gamma activity in the time window up to 80 ms after stimulus onset. In contrast, induced gamma activity as revealed by single trial analysis decreased markedly after stimulus presentation. Starting with the P3 window, induced activity recovered to baseline level for the non-target trials, whereas it remained significantly suppressed for the target responses. PMID- 9406885 TI - A comparison of (+/-)epibatidine with NMDA in releasing [3H]noradrenaline and adenosine from slices of rat hippocampus and parietal cortex. AB - We compared (+/-)epibatidine with N-methyl-D-aspartate (NMDA) in releasing adenosine and [3H]noradrenaline from slices of rat hippocampus and parietal cortex. (+/-)Epibatidine (0.1 microM) released [3H]noradrenaline but not adenosine from hippocampal slices incubated either with or without extracellular Mg2+. In contrast, NMDA (300 microM) released much more [3H]noradrenaline and also adenosine from hippocampal slices incubated in medium lacking Mg2+. (+/ )Epibatidine released neither adenosine nor [3H]noradrenaline from slices of rat parietal cortex, in contrast to NMDA which released both substances. These findings suggest that those behavioral responses to (+/-)epibatidine that are mediated by noradrenaline may involve the hippocampus but not the parietal cortex. Moreover, it seems unlikely that any of the behavioral effects of (+/ )epibatidine are mediated by adenosine release in either the parietal cortex or the hippocampus. PMID- 9406886 TI - Expression of beta-catenin and the adenomatous polyposis coli tumour suppressor protein in mouse neocortical cells in vitro. AB - Beta-catenin is known to associate with the tumour suppressor protein adenomatous polyposis coli (APC), which is highly expressed in developing brain. We have therefore investigated the distribution of beta-catenin and APC in primary cultures of mouse neocortex. Western blotting demonstrated the presence of a single beta-catenin species in our cultures. Immunocytochemistry showed that beta catenin was plasma membrane associated and concentrated in growth cones in cultured neurons. The APC tumour suppressor protein was also concentrated in growth cones. In glial cells, beta-catenin was localised at cell-cell contacts in a manner similar to that previously described in other cell types. This data suggests a role for both APC and beta-catenin in neuronal growth cones, and for beta-catenin in the formation of cell to cell contacts between glia. PMID- 9406887 TI - PAF analogues capable of inhibiting PAF acetylhydrolase activity suppress migration of isolated rat cerebellar granule cells. AB - Intracellular platelet-activating factor (PAF) acetylhydrolase in the bovine brain is a heterotrimeric enzyme composed of alpha1, alpha2 and beta subunits. The trimeric enzyme may be involved in neural cell migration, since the human homolog of the non-catalytic beta subunit is a product of the LIS-1 gene which is a causative gene for Miller-Dieker syndrome. Miller-Dieker syndrome is a form of lissencephaly that is characterized by widespread agyria of the brain and defects of neuronal cell migration. In the present study, we attempted to determine whether the catalytic activity of either the alpha1 or alpha2 subunit is required for the regulation of granule cell migration. Granule cells prepared from rat cerebellum at postnatal day 0 express all three subunit proteins (alpha1, alpha2 and beta) as determined by western blotting. Granule cell migration, which was observed in vitro on a layer coated with laminin, was effectively blocked by PAF analogs which showed PAF receptor-antagonistic activity (CV-6209 and CV-3988) and PAF receptor-agonistic activity (carbamoyl PAF). These PAF analogs also inhibited the activity of bovine brain PAF acetylhydrolase. Cell migration was restored when the inhibitors were removed by washing the treated cells with buffer, indicating that the inhibitory effect of PAF analogs is reversible. Structurally unrelated PAF antagonists (SM-12502, TCV-309 and YM-264), none of which showed any appreciable inhibitory activity against PAF acetylhydrolase, did not block granule cell migration under the same conditions. It is suggested that the catalytic activity of PAF acetylhydrolase may play a crucial role in neural cell migration. PMID- 9406888 TI - Evidence against increased oxidative DNA-damage in Down syndrome. AB - In Down syndrome (DS), oxidative DNA-damage may play a role in the pathogenesis of characteristic mental retardation and precocious dementia of Alzheimer type. We measured the oxidized nucleoside, 8-hydroxy-2'-deoxyguanosine (8-OHdG), in nuclear DNA (nDNA) isolated from four different regions of cerebral cortex and cerebellum in 10 adult DS and 10 Alzheimer's disease (AD) patients compared to normal controls. Levels of 8-OHdG in post-mortem brain tissue were investigated by means of high-performance liquid chromatography with electrochemical detection. There was no significant increase in DS and AD compared to controls in any of the brain regions. Highest amounts of 8-OHdG were in temporal cortex in DS (180.0 +/- 9.6 nmol/g wet weight tissue), AD (172.4 +/- 14.6 nmol/g wet weight tissue) and controls (183.4 +/- 12.7 nmol/g). We conclude that the results provide evidence against an increased reactive oxygen species (ROS) induced damage to nDNA in DS and AD. PMID- 9406889 TI - Early detection of optic nerve-evoked response in the superior colliculus of the neonatal rat. AB - Optic nerve-evoked responses were measured in the superior colliculus (SC) of neonatal rats in vivo from postnatal day (P) 0 to P11. At P1, a biphasic response was recorded in superficial layers and the amplitude diminished as the electrode penetrated into the deeper layers of the SC. By P2, a similar response, with a fast positive-going potential followed by a more prolonged negative potential was observed at the surface. The polarity of the response reversed as the electrode was moved into the deeper laminae of the SC. Such a reversal in the polarity of optic nerve-evoked responses resembled those observed in more mature preparations. Using current source density analysis, a single pair of source-sink could be identified following optic nerve stimulation at P2, and this changed to a more complex pattern by P11. Our results suggest that synaptic transmission in the retinocollicular pathway of the rat is functional as early as P2. PMID- 9406890 TI - Connectivity patterns of cone horizontal cells in blue acara (Aequidens pulcher, Cichlidae) reared in different light regimes. AB - Two types of cone horizontal cells were identified morphologically in the retina of a trichromatic fish by fluorescent labelling with Lucifer Yellow and confocal laser scanning microscopy. H1 cells are located adjacent to the outer plexiform layer, have large somata, small dendritic fields, and contact all cone types. H2 cells are positioned vitread to the H1 cells, have small somata, and large dendritic fields. Their dendrites invaginate the synaptic pedicles of short wavelength sensitive single cones and show a significant preference for one of the spectrally different members of the double cones, presumably the middle wavelength sensitive member. We tested the impacts of different visual environments on the development of these connectivity patterns and found minor changes induced by rearing in white light of different intensities or monochromatic blue light. PMID- 9406891 TI - Characterization of tau proteins in human neuroblastoma SH-SY5Y cell line. AB - Here we report three experimental paradigms in which tau proteins are differentially localized and expressed in human neuroblastoma cells SH-SY5Y. We found that in undifferentiated cells, tau proteins were predominantly localized in the nucleus. Western blot analysis of nuclear extracts revealed, among the others, a high molecular weight tau isoform and evaluation of tau mRNA levels showed a single tau isoform. After differentiation, tau immunoreactivity was detected only in cytosol and along neuritic processes. The high molecular weight tau isoform disappeared and an additional tau mRNA species was detected. Treatment of differentiated cells with doxorubicin or okadaic acid resulted in an increase of tau immunoreactivity and in a subsequent cell loss. Our results indicate that both subcellular localization and pattern of expression of tau proteins vary depending on the developmental and functional state of the cells, thus suggesting different roles in cell function. PMID- 9406892 TI - The effect of naturally occurring fragments of galanin message-associated peptide on spinal cord excitability in rats. AB - Galanin message-associated peptide (GMAP), a 60 amino acid fragment of galanin precursor protein, is present in dorsal root ganglion cells and upon intrathecal (i.t.) administration influences the spinal nociceptive flexor reflex in rat in a complex manner. The present study assessed the effects on spinal cord excitability of N-terminal fragment GMAP (1-30) and C-terminal fragments GMAP (34 60) or GMAP (35-60), which were formed from GMAP following enzymatic degradation. The effect of the fragments was compared with the effects of the complete peptide sequence. The GMAP fragments slightly facilitated the flexor reflex and dose dependently blocked hyperexcitability following C-fiber conditioning stimulation. The potency of the blocking effect of GMAP (1-30) was comparable to GMAP (1-60) and was one order of magnitude higher than the potency of the C-terminal fragments. The results indicated that both naturally formed N- and C-terminal fragments of GMAP are pharmacologically active and produce effects which are similar to the full peptide sequence. PMID- 9406893 TI - Stimulated release of the beta-amyloid protein of Alzheimer's disease by normal human platelets. AB - The circulatory system is a potential source of the beta-amyloid protein (A beta) of ageing and Alzheimer's disease (AD), platelets accounting for the bulk of A beta immunoreactivity detectable in blood. Evidence for the release of A beta by platelets, however, has not been reported. Platelets from normal donors were therefore stimulated with collagen to establish if A beta immunoreactive material is released on activation. For comparison, the release of the platelet monoamines, serotonin (5-HT) adrenaline (Adr) and noradrenaline (NA) was also measured. Like the monoamines, collagen-induced A beta release was concentration dependent, maximal stimulated release exceeding basal efflux by 184%. Collagen EC50 values for A beta release were similar to those for Adr and NA (3.6 +/- 0.6, 3.4 +/- 0.6 and 3.3 +/- 0.2 microg/ml collagen, respectively) but not 5-HT (9.8 +/- 1.9 microg/ml). These data provide the first evidence that platelets release A beta immunoreactive material on stimulation and may indicate that A beta, Adr and NA reside in the same subcellular compartment. PMID- 9406894 TI - Antimicrobial screening of selected medicinal plants from India. AB - From the Indian traditional medicines 78 plants were selected on the basis of their use in the treatment of infectious diseases. Different concentrations of 80% ethanol extracts were tested, using the agar dilution method, against four bacteria: Bacillus subtilis, Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa and, using the agar-well diffusion method, against two fungi: Candida albicans and Aspergillus niger. In the lowest tested concentration of 1.6 mg/ml, 10% of the plant extracts were active; 44% in a concentration of 6.25 mg/ml and 90% of the plant extracts were active against at least two bacteria in a concentration of 25 mg/ml. Only 13% of the plant extracts were active against at least one fungus in a concentration of 50 mg/ml. PMID- 9406895 TI - Antimicrobial study of the resinous exudates and of diterpenoids and flavonoids isolated from some Chilean Pseudognaphalium (Asteraceae). AB - The antimicrobial properties of the resinous exudates from twigs and leaves of four Chilean species of Pseudognaphalium: P. viravira, P. robustum, P. heterotrichium and P. cheiranthifolium were tested against six gram negative bacteria and five gram positive bacteria. The extracts share similar antimicrobial activities against the gram positive bacteria. The antimicrobial activity correlated very well with the presence in the resinous exudates of ent 16-kauren-19-oic acid and to a lesser extent with the presence of ent-9(11),16 kauradien-19-oic. Introduction of an hydrophilic 3 beta-OH drastically reduced the antimicrobial activity of these compound. The activity was not correlated with the flavonoid content of those resinous exudates. PMID- 9406896 TI - Immunopotentiating compounds from Tinospora cordifolia. AB - The active principles of Tinospora cordifolia a traditional Indian plant were found to possess anticomplementary and immunomodulatory activities. Syringin (TC 4) and cordiol (TC-7) inhibited the in vitro immunohaemolysis of antibody-coated sheep erythrocytes by guinea pig serum. The reduced immunohaemolysis was found to be due to inhibition of the C3-convertase of the classical complement pathway. However, higher concentrations showed constant inhibitory effects. The compounds also gave rise to significant increases in IgG antibodies in serum. Humoral and cell-mediated immunity were also dose-dependently enhanced. Macrophage activation was reported for cordioside (TC-2), cordiofolioside A (TC-5) and cordiol (TC-7) and this activation was more pronounced with increasing incubation times. PMID- 9406897 TI - Determination of iron content in different parts of herbs used traditionally for anaemia treatment in East Africa. AB - The iron content in different parts of eight plants traditionally used to treat anaemia in Eastern Africa was determined using atomic absorption spectrophotometry. Extracts were made of plant samples using both wet and dry oxidation procedures. Results obtained from both procedures agree significantly and the average of both methods was taken as the iron content in each plant part. In most cases, the values obtained for the rootbark had a higher total iron content than the corresponding leaves and stembark. The prominent iron contents of 35.69 and 35.21 mg/100 g were found in the rootbark of Bridelia cathartica and Lannea stuhlmannii, respectively. The iron content of the decoctions prepared in the traditional way was low. However, the therapeutic potential of the herbs cannot be established on the basis of available iron content alone as other factors play a role in the absorption of iron in the body. PMID- 9406898 TI - Effects of Sangre de Drago from Croton lechleri Muell.-Arg. on the production of active oxygen radicals. AB - The total reactive antioxidant potential (TRAP) of 'Sangre de Drago' from Croton lechleri (Euphorbiaceae) was determined by monitoring the intensity of luminol enhanced chemiluminescence enhanced by peroxyl radicals derived from thermolysis of 2,2'-azo-bis(2-amidinopropane). The TRAP index was calculated as 935.4 +/- 141 microM, measured as equivalents of Trolox concentration. On the other hand, the additive incorporation of lower concentrations yielded an instantaneous increase in chemiluminescence, suggesting a prooxidant activity at these levels. DNA sugar damage induced by Fe(II) salts was also used to determine the capacity of the latex to suppress hydroxyl radical-mediated degradation of DNA. As in the case of luminol enhanced chemiluminescence, Sangre de Drago was highly effective in reducing oxidation of DNA at higher concentrations, but showed an increase in the production of TBARS at lower doses, as compared to the control. Finally, antioxidant activity was tested using hydroperoxide-initiated chemiluminescence in rat liver homogenates, and the latex showed an increase in light emission, suggesting the presence of prooxidant compounds. PMID- 9406899 TI - In vivo protective role of Koflet (an ayurvedic preparation) against cellular toxicity caused by CCl4 and flyash. AB - Swiss albino rats were treated in groups with CCl4, and flyash to induce cellular toxicity in the lungs and trachea. Animal groups received treatment of Koflet (K) with CCl4 (7 days) and with flyash (30 days); their general health and biochemical parameters were studied and used as an indication of cellular injuries. A significant loss was observed in body weight and food consumption in animals given only CCl4 or flyash, while simultaneous treatment with K resulted in a non significant alteration from normal control groups. Enzyme (alkaline phosphatase, Ca2+ -Mg2+ -ATPase, glutamic-oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT)) activities were estimated in tissue homogenate preparation of lung, trachea and serum, which showed no significant change except for GPT activity as compared to control animals which received CCl4 or flyash with K. Similarly lung, trachea and serum contents of carbohydrate, protein, sialic acid, serum protein, serum cholesterol were estimated and it was found that alteration caused by CCl4, or flyash becomes almost non-significant compared to that of the control after the treatment of K, except for carbohydrate and serum cholesterol values. The animal group which was only treated with K did not show any significant alteration in their biochemical markers or injuries, except for cholesterol. PMID- 9406900 TI - Vasodilating and hypotensive effects of fangchinoline and tetrandrine on the rat aorta and the stroke-prone spontaneously hypertensive rat. AB - Comparative studies of the effects of tetrandrine (TET) and fangchinoline (FAN), two major components of the Radix of Stephannia tetrandrae, on vasodilations and on calcium movement in vascular smooth muscle, and studies of hypotensive effects on stroke-prone spontaneously hypertensive rats (SHRSP) were performed in the following experiments. TET and FAN inhibited high K+ (65.4 mM) and induced sustained contraction in the rat aorta smooth muscle strips. IC50 values for TET and FAN were 0.27 +/- 0.05 microM (n = 6) and 9.53 +/- 1.57 microM (n = 6), respectively, and this inhibition was antagonized by increasing the Ca2+ concentration in the medium. The IC50 of TET for norepinephrine (NE)-induced contraction (0.86 +/- 0.04 g) was 3.08 +/- 0.05 microM (n = 4), and the IC50 of FAN for NE-induced contraction (0.88 +/- 0.07 g) was 14.20 +/- 0.40 microM (n = 4). At the molecular level, radiolabelled 45Ca2+ uptake tests revealed that TET and FAN also inhibited high K+ (65.4 mM) and 1 microM NE-stimulated Ca2+ influx in rat aorta strips at the maximal concentration was needed to inhibit the contraction. TET (3 mg/kg) and FAN (30 mg/kg) administered by intravenous (i.v.) bolus injection also lowered the mean arterial pressure (MAP) significantly during the period of observation in conscious SHRSP, respectively. These results showed that TET was more potent than FAN in blocking calcium channels and antihypertensive activity. PMID- 9406901 TI - Antinociceptive effects of Trigonella foenum-graecum leaves extract. AB - There are some reports concerning the antinociceptive effects of the plant Trigonella foenum-graecum (TFG) in Iranian traditional medicine. Because of the side effects of nonsteroidal anti-inflammatory and antinociceptive drugs, and in search for more potent and less harmful compounds, we tried to study the antinociceptive effects of TFG leaves by using tail-flick and formalin tests. Intraperitoneal (i.p.) administration of 500 mg/kg of TFG extract and 100 and 300 mg/kg of sodium salicylate (SS), as a positive control, did not show any effect in the tail-flick test, but the 1000 and 2000 mg/kg of the extract produced significant increase in the tail-flick latency. SS (300 mg/kg, i.p.) induced antinociception in the second phase of the formalin test. TFG (500 mg/kg, i.p.) demonstrated antinociception only in the first phase, but 1000 and 2000 mg/kg, i.p. doses alleviated the pain in both phases. Preliminary LD50 of the extract was very close to 4000 mg/kg, i.p. We conclude that: (1) the extract of TFG leaves produces antinociceptive effects through central and peripheral mechanisms; (2) the antinociceptive effects of 2000 mg/kg of the extract was more potent than 300 mg/kg of SS. PMID- 9406902 TI - Antihepatotoxic activity of Cichorium intybus. PMID- 9406904 TI - Psychosocial and physical correlates of chronic depression. AB - This study used a case-control design to address differences in psychosocial, physical and clinical profiles between subjects who presented with a chronic index episode of major depression and those who presented with a non-chronic index episode. Subjects were adult patients participating in the Duke University Mental Health Clinical Research Center (MHCRC) for the Study of Depression in Later Life. Cases (N = 88) who reported duration of depressive symptoms lasting > or = 24 months at enrollment were compared to controls (N = 354) who reported symptoms lasting 1-12 months. The groups were compared with respect to selected demographic and clinical variables, physical function deficits, medical comorbidity, social support constructs and number of recent stressful life events. Social support and physical health were more relevant to chronicity of major depressive illness than were severity of illness or family history. Older age (> 60 years) intensified the deleterious effect of recent negative life events and reduced the deleterious effect of functional impairment on chronic major depression. These findings require special emphasis where treatment for chronic major depression is divorced from considerations of the social environment and functional capacity. PMID- 9406905 TI - Relationships between changes in symptom ratings, neurophysiological test performance and quality of life in schizophrenic patients treated with clozapine. AB - This study examined the correlations between reduction in symptoms, changes in neuropsychological test performance and improved quality of life in 19 schizophrenic outpatients treated with clozapine. Reduction in both negative symptoms and general psychopathology was associated with a better quality of life. Some improvement in neuropsychological test performance was found, with a variable pattern of association with change in psychopathology. Improved verbal fluency was associated with reduction in negative symptoms, while improved verbal concept formation was associated with reduction in positive symptoms. PMID- 9406903 TI - Pharmacological investigation of Cassia italica. AB - In the present study, the ethanolic extract of the whole plant parts (root, stem leaves and pods) of Cassia italica (Mill.) Lam. ex F.W. Ander (Leguminosae) was investigated for bioactivities: namely anti-inflammatory, antipyretic, analgesic and prostaglandin (PG) release by rat peritoneal leucocytes, antineoplastic and antiviral. In rats, the extracts reduced carrageenin-induced paw swelling (100 mg/kg bw-31%) and fever (100 mg/kg bw-37%). The extract showed weak effect on writhing induced by acetic acid. A dose-dependent inhibition of PG release effect was observed using rat peritoneal leucocytes. PMID- 9406906 TI - Normalizing the crossover effect: enhancement of cognitive attentional processing in schizophrenia. AB - Thirty male schizophrenic subjects and 20 male control subjects were administered both the classic Rodnick and Shakow reaction time (RT) procedure and a modified experimental procedure using a balanced design in an attempt to enhance cognitive attentional processing and inhibit the crossover effect in schizophrenic patients. The experimental procedure increased the number of predictable trials, provided more information about the task and presented the regular trials in a simple ascending order. Analysis of the RT data showed the typical early schizophrenic crossover and late normal crossover on the standard task, while schizophrenic patients in the experimental condition showed a new finding of statistically significant normalization of the early crossover effect. The results support Shakow's theory of attentional set deficit in schizophrenia and in conjunction with previous findings suggest that separate neuropsychological mechanisms of cognitive preparatory attention and a more automatic form of vigilance attention may, respectively, underlie the regular and irregular procedures of the traditional RT task. PMID- 9406908 TI - Beta-endorphin, adrenocorticotropic hormone and cortisol secretion in abstinent alcoholics. AB - The circadian secretion of beta-endorphin, adrenocorticotropic hormone (ACTH), and cortisol was evaluated in 14 non-cirrhotic alcoholic men after 7 and 28 days of abstinence and in 12 sex- and age-matched normal subjects. A significant decrease in plasma levels of beta-endorphin, reduced ACTH levels, and increased cortisol levels were observed in samples taken at 08.00 h, 12.00 h, 18.00 h, and 23.00 h both after 7 and 28 days of abstinence. These data suggest the presence of a strong negative feedback on pro-opiomelanocortin synthesis by cortisol hypersecretion in abstinent alcoholics, which might be due to long-term stimulation of adrenal function by alcohol. The decreased plasma beta-endorphin levels might predispose to relapse in alcohol abuse. PMID- 9406909 TI - Validation of a computerized version of the temperament and character inventory (TCI) in psychiatric inpatients. AB - We wrote a Basic program for Macintosh and IBM-compatible computers in order to administer the 226 questions of the French Temperament and Character Inventory (TCI) automatically, and to obtain a personality profile instantaneously. Validity was assessed by comparing the results of 32 psychiatric inpatients who used this program and the paper-and-pencil form of the TCI over an interval of 4.8 days on average. No acceptability problem was met with the computerized version. Correlation coefficients between computer and paper-and-pencil scores varied from 0.78 to 0.91 for temperament dimensions, and from 0.71 to 0.82 for character dimensions. The mean scores of the two versions were not significantly different. These results suggest that a computerized version of the TCI can be a reliable and acceptable instrument for the assessment of personality dimensions in psychiatric patients. PMID- 9406907 TI - Serotonergic dysfunction in addiction: effects of alcohol, cigarette smoking and heroin on platelet 5-HT content. AB - The impact of ethanol, cigarette smoking and heroin on serotonin function was evaluated, first in alcoholics during chronic ethanol intoxication and in opiate addicts after long-term heroin consumption, and secondly in both patient groups after detoxification treatment (i.e. a short-term abstinence of 8 days). Our results showed that the 5-hydroxytryptamine (5-HT) content in platelets was: (1) increased in the subgroup of anti-social alcoholics; (2) transiently and differently altered in alcoholics compared to opiate addicts; and (3) lowered in drinking alcoholics and normal in alcoholics who were drinking as well as smoking (that may occur via MAO-B inhibition by smoke). The findings indicate that alterations of the peripheral and possibly the central serotonin system may occur as predisposing factors for alcoholism in individuals with anti-social traits; they may also have some impact on the progression of alcoholism due to its lowered function during chronic ethanol intoxication that is substantially modified by smoking. PMID- 9406910 TI - Over the mainstream: diagnostic requirements for biological psychiatric research. AB - Progress in biological psychiatry is contingent on progress in neurobiology and on research into proper characterisation and assessment of abnormal behavior. Advances in neurobiology are rapid and steady; diagnostic research does not keep pace. On the contrary, the diagnostic approach seems solidified, as today's basic premises are uncritically accepted. The diagnostic requirements for meaningful biological psychiatric research are discussed and contrasted with present-day diagnostic practices. Serious reflection on the state of psychiatric diagnosing is urgently needed. PMID- 9406911 TI - Multiple signaling pathways direct the initiation of tyrosine hydroxylase gene expression in cultured brain neurons. AB - Previous studies have demonstrated that the synergistic interaction of acidic fibroblast growth factor (aFGF) and a second co-activator molecule can novelly induce expression of the CA biosynthetic enzyme tyrosine hydroxylase (TH) in non TH expressing neurons of the striatum. Several co-activators have been identified, including substances present in L6 muscle cell extract (X. Du et al., J. Neurosci. 14 (1994) 7688-7694) catecholamines, such as dopamine (DA) (X. Du and L. Iacovitti, J. Neurosci. 15 (1995) 5420-5427; X. Du et al., Brain Res. 680 (1995) 229-233) and activators of protein kinase C (PKC) such as TPA (X. Du and L. Iacovitti, J. Neurochem. 68 (1997) 564-569). In the present study, we investigated whether activators of the protein kinase A (PKA) pathway also serve as effective co-activators of aFGF in the induction of TH gene expression. In addition, the combinatorial effects of the various TH-inducing agents were also evaluated. We found that, as with other co-activating molecules, the PKA stimulants IBMX and forskolin had no TH-inducing capacity when administered alone. However, co-treatment of 10 ng/ml aFGF with either (250 microM) IBMX or (10 microM) forskolin resulted in the novel expression of TH in 25% of plated neurons. The number of TH-expressing neurons was increased to 55% in aFGF-treated cultures co-incubated with aFGF and both (250 microM) IBMX and (10 microM) forskolin. Time course studies indicated that TH induction was rapid (peaking within 24 h) and enduring (lasting 4 days in culture). Induction of TH by aFGF and IBMX/forskolin was partially blocked by inhibitors of protein kinase, such as H7, H8 and H89, as well as pretreatment with protein (cyclohexamide) or RNA synthesis (amanitin and actinomycin D) inhibitors. The concomitant addition of combinations of co-activator molecules (DA, TPA and IBMX/forskolin) and aFGF resulted in the additive induction of TH. Maximal expression of TH (80% of striatal neurons) was accomplished when cultures were treated with aFGF and all co-activator molecules simultaneously. Our results suggest that there are multiple ways to signal the initiation of the TH gene, each of which requires the synergy of specific growth factors and either DA, PKA or PKC pathway activators. Since only the combination of growth factor and all co-activators together produces maximum TH induction, each molecule may signal a unique intracellular pathway which converges at targets on the TH gene. PMID- 9406912 TI - Molecular analysis of the monomeric GTP-binding proteins of oligodendrocytes. AB - Vesicle transport plays an important role in the formation of myelin. Transport of proteins, including proteolipid protein and myelin associated glycoprotein, from their site of synthesis in the endoplasmic reticulum in the perikaryon of the oligodendrocytes, to myelin, takes place via carrier vesicles. The mechanisms that regulate vesicle transport in oligodendrocytes are largely unknown. The presence of monomeric GTP-binding proteins in myelin and oligodendrocytes suggested the hypothesis that these proteins participate in the regulation of vesicle transport. In an attempt to identify the Rab and Rho GTP-binding proteins present in oligodendrocytes, a cDNA library specific for these proteins was generated using a reverse transcriptase-polymerase chain reaction (RT-PCR) approach. Twelve different clones containing sequences that coded for members of the Rab and Rho families of GTP-binding proteins were isolated. This group includes Rab1, -1b, -2, -5b, -5c, -7, -8, -12, -14, -23 and Rho A. One additional clone revealed a novel cDNA sequence. Analysis of the effector loop motif indicated that this sequence encodes for a member of the Rab family. We refer to this new sequence as Rab0. Comparison of Rab0 with the most similar rat Rab sequences, Rab 14 and Rab 22, and with a recently cloned human Rab22b, showed a 71%, 72% and 94% identity, respectively. By RT-PCR analysis the Rab0 mRNA was found to be mainly expressed in oligodendrocytes and to a lesser extent in oligodendrocyte precursors, astrocytes and microglia. Moreover, the highest levels of Rab0 mRNA were observed in areas of the brain that are heavily myelinated. Rab0 mRNA was also detected in other tissues such as kidney, liver, skeletal muscle. These data provide initial evidence regarding signal transduction pathways that regulate intracellular transport in oligodendrocytes. PMID- 9406913 TI - Activation of CPP-32 protease in hippocampal neurons following ischemia and epilepsy. AB - Recent in vitro studies indicate an involvement of members of the interleukin 1beta converting enzyme (ICE) family of proteases in programmed neuronal cell death. Cell death of hippocampal neurons in animal models of cerebral ischemia and epilepsy shows morphological features of apoptosis and can be prevented by administration of protein synthesis inhibitors suggesting that de novo synthesis of components of the cell death program is necessary for neuronal apoptosis. In the present study we demonstrate by in situ hybridization analysis that expression of CPP-32, an ICE-related protease, is significantly upregulated in CA1 hippocampal neurons following global ischemia induced by cardiac arrest and in hippocampal neurons of the CA3/CA4 region after kainate-mediated epilepsy, respectively. Moreover, an increase in CPP-32-like proteolytic activity was detected in hippocampal extracts 24 h after ischemia using the fluorogenic CPP-32 substrate Ac-DEVD-AMC. Activation of CPP-32 clearly preceded cell death of hippocampal neurons as assessed by in situ end-labelling of nuclear DNA fragments. These results indicate that CPP-32 protease may be activated at both the transcriptional and post-translational level during neuronal apoptosis and that activation correlates with the selective vulnerability of hippocampal pyramidal neurons to ischemic and epileptic insults. PMID- 9406914 TI - Controlled targeting of tyrosine hydroxylase protein toward processes of locus coeruleus neurons during postnatal development. AB - Dendrites of locus coeruleus (LC) neurons laying within the pericoerulean neuropil (PCA) organize the major site where tyrosine hydroxylase (TH) is present throughout postnatal development. Those dendrites constitute the neuronal compartment in which TH levels increase beyond postnatal day (P) 21 or after RU24722-induced TH expression. Distal LC dendrites are present in the PCA by at least P20 but are devoid of TH and can rapidly accumulate TH protein when gene induction is triggered. Contrasting with the increase in TH levels within LC perikarya and dendrites, TH-mRNA concentration remains constant in LC perikarya from P4 to P42. Thus, supposing TH synthesis and degradation are also constant, any change in TH levels targeted toward axons might be balanced by a shift in the TH deposition within LC dendrites. This mechanism may be crucial in functions that the different processes of LC neurons have at critical steps of postnatal ontogeny. PMID- 9406915 TI - Identification of cell type-specific promoter elements associated with the rat tyrosine hydroxylase gene using transgenic founder analysis. AB - Transcriptional regulatory elements capable of directing transgene expression to individual cells are powerful tools for manipulating a given CNS circuit. Delineating these elements via traditional transgenic analysis is both costly and labor intensive. Here we have used the rat tyrosine hydroxylase (TH) promoter as a model to describe and validate the use of founder animals for systematic promoter studies. No significant differences were found when data obtained from founder animals expressing a 6.0 kb TH promoter directing LacZ were compared with animals derived from an analogous transgenic line. Subsequent studies with founder animals expressing beta-galactosidase directed by various lengths of rat TH promoter revealed different patterns of expression. Specifically, a locus coeruleus regulatory domain was localized between 3.4 and 6.0 kb of the rat TH promoter, a hypothalamic regulatory domain between 2.5 and 3.4 kb and a brainstem regulatory domain between 0.8 and 6.0 kb. At least one element of a midbrain specific regulatory domain was within 2.5 kb of the transcriptional start site. Olfactory bulb specific elements however appeared to reside outside of the sequences tested. Specific patterns of ectopic gene expression were also observed suggesting the presence of negative regulatory elements. Thus, TH appears to be regulated in a complex modular fashion by both positive and negative regulatory elements. Taken together, this study demonstrates the feasibility and reliability of founder analysis for promoter studies of genes expressed in complex spatial and temporal patterns. PMID- 9406916 TI - Effects of angiotensinogen antisense oligonucleotides on fluid intake in response to different dipsogenic stimuli in the rat. AB - The role of centrally synthesised angiotensinogen in neural mechanisms subserving water drinking in rats was investigated by injecting antisense oligonucleotides complementary to rat angiotensinogen mRNA into the brain with the aim of inhibiting cerebral angiotensinogen synthesis. Phosphorothioate antisense oligonucleotides (18 mer) encompassing the translation start codon were injected into the lateral ventricle of rats and their responses to a number of dipsogenic stimuli tested. These were: intracerebroventricular (i.c.v.) renin, i.c.v. angiotensin II, i.c.v. carbachol, subcutaneous isoproterenol, intravenous hypertonic saline, water deprivation for 24 h or subcutaneous injection of polyethylene glycol. Antisense treatment significantly reduced (by approximately 50%) the volume of water drunk in response to i.c.v. injection of renin or subcutaneous isoproterenol, but did not reduce water intake elicited by the other dipsogenic stimuli. The i.c.v. administration of mismatch, scrambled or sense oligonucleotides did not inhibit water intake. These data suggest that centrally produced angiotensinogen may have a role in neural pathways subserving isoproterenol-induced drinking. PMID- 9406917 TI - Evidence for autocrine inhibition of gonadotropin-releasing hormone (GnRH) gene transcription by GnRH in hypothalamic GT1-1 neuronal cells. AB - To examine whether an ultrashort feedback mechanism of gonadotropin-releasing hormone (GnRH) operates at the level of gene transcription, we studied the effects of GnRH analogs on GnRH promoter activity and GnRH mRNA level in hypothalamic GT1-1 neuronal cells. Treatment of GT1-1 cells with buserelin, a GnRH agonist, or native GnRH for 24 h significantly decreased GnRH promoter activity and its mRNA level, whereas that with GnRH antagonists, antide or [D Phe2,D-Ala6]-GnRH, showed no effect. The inhibitory effects of buserelin on GnRH gene transcription and GnRH mRNA level were dose-related, and a significant inhibition was observed in cells treated with buserelin at concentrations higher than 0.1 microM. Time-course experiments showed that significant decreases in GnRH promoter-driven luciferase activity and GnRH mRNA level were observed within 12 h and sustained up to 48 h. Moreover, treatment with GnRH agonist for 12 h significantly decreased the transcription rate of the mouse GnRH gene, as revealed by nuclear run-on transcription assay. The promoter analysis with the 5' deletional constructs demonstrated that cis-acting elements important for GnRH autoregulation by GnRH agonist reside within -854 bp upstream from the transcription start site. These data clearly demonstrate that GnRH can exert autocrine regulation at the level of GnRH gene transcription. PMID- 9406918 TI - Selective increase in somatostatin mRNA expression in human basal ganglia in Parkinson's disease. AB - Levels of the neurotransmitter somatostatin (SS) have previously been shown to be reduced in the cortex and hippocampus of demented parkinsonian patients and patients with Alzheimer's disease. In situ hybridisation histochemistry (ISHH) was performed with an 35S tail-labelled oligonucleotide DNA probe to human SS mRNA, to examine its expression within the striatum, medial medullary lamina (MML) and reticular thalamic nucleus in Parkinson's disease (PD) and in matched controls. A chronic unilaterally MPTP-lesioned L-DOPA-naive primate model was also examined for comparison of SS mRNA expression with that in human L-DOPA treated PD subjects. Quantitation of SS mRNA expression on emulsion dipped sections revealed a significant increase (82%) in the MML of the globus pallidus in PD (56.5 microm2 of silver grain/cell, n = 9 cases) compared to controls (26.3 microm2/cell, n = 13 cases, p < 0.01, Student's t-test), paralleling the increase previously observed by this group for NOS mRNA. SS mRNA expression was higher in the dorsolateral than ventromedial putamen in controls (p < 0.001; DL: 24.89 +/- SEM 1.35; VM: 17.96 +/- SEM 2.63; n = 14) but this gradient was lost in PD cases (p > 0.05; DL: 22.68 +/- 1.94; VM: 22.17 +/- 2.94; n = 10). These findings suggest specific modification of basal ganglia SS-ergic pathways in PD. PMID- 9406919 TI - BDNF protection of auditory neurons from cisplatin involves changes in intracellular levels of both reactive oxygen species and glutathione. AB - Previous studies have shown that brain derived neurotrophic factor (BDNF) can protect auditory neurons from cisplatin toxicity in vitro. To explore the mechanism of BDNF mediated neuronal protection sequential confocal microscopic sampling of auditory neurons measured intracellular levels of reactive oxygen species (ROS) in response to withdrawal of BDNF supplementation, cisplatin exposure, and BDNF protection from cisplatin damage in normal and oxidative stress states. Additionally, we examined intraneuronal levels of the free radical scavenger glutathione (GSH) in response to withdrawal of BDNF. Withdrawal of BDNF resulted in increased production of ROS and decreased survival of auditory neurons. Levels of GSH within neurons increased after BDNF withdrawal, and this increase was shown to lag behind the production of ROS. Auditory neurons in cultures supplemented with BDNF and exposed to cisplatin showed significantly lower levels of ROS and increased survival compared to neurons in unprotected, cisplatin exposed cultures. Neurons treated with buthionine sulfoximine (an inhibitor of GSH synthesis), supplemented with BDNF, and exposed to cisplatin showed significantly higher intracellular levels of ROS compared to the neurons in BDNF supplemented cultures exposed to cisplatin. These results suggest that intracellular levels of ROS play an important role in cisplatin induced cell death of auditory neurons and that production of ROS can be ameliorated through supplementation with BDNF. GSH appears to mediate BDNF protection of these neurons from cisplatin induced ROS and subsequent damage. PMID- 9406920 TI - Effects of pentylenetetrazol-induced status epilepticus on c-Fos and HSP72 immunoreactivity in the immature rat brain. AB - Pentylenetetrazol (PTZ)-induced status epilepticus (SE) leads to acute and long term metabolic decreases in specific brain regions of rats at 10 (P10) or 21 days after birth (P21). These decreases are not related to apparent neuronal damage. Therefore, to better understand the neuronal activation and stress response to PTZ in immature rats, we mapped the expression of c-Fos and of the 72 kDa heat shock protein (HSP72) in the same model of severe SE induced by the repetitive i.p. injections of subconvulsive doses of PTZ. Rats were sacrificed either at 2 or 24 h after the onset of SE in order to reveal c-Fos immunoreactivity, and at 24 and 72 h for HSP72 expression. Hematoxylin-eosin staining was performed at 24, 72 and 144 h after SE. The expression of c-Fos at 2 h after SE was more marked at P21 than at P10 and was prominent at both ages in the hippocampal dentate gyrus, cerebral cortex and amygdala. Some immunoreactivity was also present in the hypothalamus, thalamus and a few brainstem and cerebellar regions at both ages. There was a good relation between the regions expressing c-Fos and those exhibiting acute metabolic decreases at P21. Conversely, PTZ seizures did not lead to any expression of c-Fos at 24 h after SE or of HSP72 at 24 or 72 h at any age. Cell density was not affected by PTZ-induced SE at any age and at any time. These results suggest that c-Fos is a useful marker of neuronal activation induced by severe and prolonged seizures in the immature brain. The lack of HSP72 and of late c-Fos expression likely reflect the absence of neuronal damage in this model of PTZ-induced SE in the immature rat. PMID- 9406921 TI - A truncated Reelin protein is produced but not secreted in the 'Orleans' reeler mutation (Reln[rl-Orl]). AB - Reelin is the protein defective in reeler mutant mice [I. Bar, C. Lambert de Rouvroit, I. Royaux, D.B. Krizman, C. Dernoncourt, D. Ruelle, M.C. Beckers, A.M. Goffinet, A YAC contig containing the reeler locus with preliminary characterization of candidate gene fragments, Genomics 26 (1995) 543-549; G. D'Arcangelo, G.G. Miao, S.C. Chen, H.D. Soares, J.I. Morgan, T. Curran, A protein related to extracellular matrix proteins deleted in the mouse mutant reeler, Nature 374 (1995) 719-723; S. Hirotsune, T. Takahara, N. Sasaki, K. Hirose, A. Yoshiki, T. Ohashi, M. Kusakabe, Y. Murakami, M. Muramatsu, S. Watanabe, K. Nakao, M. Katsuki, Y. Hayashizaki, The reeler gene encodes a protein with an EGF like motif expressed by pioneer neurons, Nature Genet. 10 (1995) 77-83]. In the Orleans allele of reeler (symbol: Reln[rl-Orl]), a 220 nucleotide deletion is present in the 3' region of the Reelin message, resulting in a frame shift with production of a predicted protein amputated from its C-terminal amino acids. In this study, we first show that the predicted truncated protein indeed exists in Orleans reeler mice, using several anti-Reelin antibodies. Three antibodies are directed against epitopes located in the N-terminal region of the protein, namely: monoclonal antibody CR-50 [M. Ogawa, T. Miyata, K. Nakajima, K. Yagyu, M. Seike, K. Ikenaka, H. Yamamoto, K. Mikoshiba, The reeler gene-associated antigen on Cajal-Retzius neurons is a crucial molecule for laminar organization of cortical neurons, Neuron 14 (1995) 899-912] (epitope region between Reelin residues 251-407), monoclonal antibody G10 (epitope located between amino acids 199 and 244) and the polyclonal antipeptide rp4 (positions 381-399). A fourth antibody, antipeptide rp5, reacts with the C-terminal (3443-3461) Reelin sequence. In normal embryos, all four antibodies stained cells in the marginal zone with features of Cajal-Retzius cells. While N-terminal specific antibodies detected Reelin immunoreactivity in mouse embryos homozygous for the reeler Orleans mutation, no staining was obtained with the rp5 antibody, showing the presence of a truncated protein. Moreover, although Reelin could be detected at the surface of living Cajal-Retzius cells of normal mice, it was not revealed after vital staining of embryonic cortex from Orleans reeler mice. These results indicate that the C-terminal region of Reelin is essential for its secretion and suggest that the Orleans reeler phenotype is due to defective Reelin secretion rather than to secretion of an inactive protein. PMID- 9406922 TI - Adenovirus-mediated expression of AMPA-type glutamate receptor channels in PC12 cells. AB - The alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type glutamate receptor channels are expressed ubiquitously in brain neurons and mediate fast excitatory neurotransmission. They are composed of four subunits, GluR1, GluR2, GluR3 and/or GluR4. We constructed recombinant adenoviruses encoding rat AMPA receptor subunit cDNAs, GluR1 (AxCAGluR1) and silently mutated GluR2 (AxCAGluR2X) with modified chicken beta-actin promoter and cytomegalovirus immediate-early enhancer. Using these adenoviral vectors, we transferred the GluR1 and GluR2 genes into PC12 cells that possess no functional AMPA receptor channels. PC12 cells infected with these viruses expressed GluR1 and GluR2 RNAs. Immunoblot analysis indicated that the expressed GluR1 and GluR2 proteins were equivalent to those of the rat brain. Functional expression of the AMPA receptor channels was examined using the whole-cell patch clamp technique. In AxCAGluR1 infected cells, the current-voltage (I-V) relationship of response to kainate, a non-desensitizing agonist of AMPA receptors, exhibited a strong inward rectification, indicating the formation of functional GluR1-homomeric channels. In cells infected with both AxCAGluR1 and AxCAGluR2X, the I-V relationship of kainate responses exhibited an outward rectification, indicating the formation of heteromeric GluR1/R2 channels. Immunocytochemical analysis revealed that the AMPA receptor subunit genes were transferred in more than 95% of the infected PC12 cells. PMID- 9406923 TI - Estrogen regulates galanin but not tyrosine hydroxylase gene expression in the rat locus ceruleus. AB - The neuropeptide galanin (GAL) is coexpressed by the majority of noradrenergic neurons in the rat locus ceruleus (LC) and may function as an inhibitory modulator of noradrenergic transmission. Because estrogen has been shown to induce GAL expression in other brain regions and modulate noradrenergic transmission, we used in situ hybridization histochemistry to assess the effects of chronic estrogen treatment on GAL and tyrosine hydroxylase (TH) gene expression in the LC of ovariectomized female rats. We found that GAL mRNA levels were significantly elevated in rats implanted with a Silastic capsule containing estradiol compared to sham-implanted controls. Both the average optical density (P < or = 0.05) and the labelling area (P < or = 0.007) differed significantly between the groups. In contrast, TH gene expression measured in alternate brain sections did not differ between the groups. If GAL functions as an inhibitory modulator of noradrenergic transmission as postulated, these findings suggest that chronic estrogen treatment could reduce the noradrenergic tone of the brain in the absence of significant alterations in TH expression by enhancing the level of cosecreted GAL. PMID- 9406924 TI - Ibuprofen: effect on inducible nitric oxide synthase. AB - Treatment with ibuprofen and other non-steroidal anti-inflammatory drugs (NSAIDS) has been reported to decrease the incidence as well as slow down the progression of Alzheimer's disease. Understanding the mechanism of this therapeutic effect would provide a target for development of drugs which may be devoid of side effects observed with NSAIDs. In addition to inhibiting cyclooxygenase (COX), the NSAIDs have recently been shown to decrease inducible nitric oxide synthase (iNOS) activity. Ibuprofen and other NSAIDs had no direct effect on catalytic activity of iNOS, but decreased levels of iNOS mRNA. The mechanism of action of ibuprofen on reduction of iNOS activity has been further investigated in the present study using rat primary cerebellar glial cell cultures. In addition, the effect of ibuprofen on COX mRNA expression and prostaglandin formation was also studied. Glial cells treated with E. coli lipopolysaccharide (LPS) and interferony (INFgamma) for 16 h expressed iNOS and COX. Ibuprofen did not directly inhibit iNOS activity. However, when ibuprofen was incubated at the same time with LPS and INFgamma for 16 h, enzyme activity was reduced, with an IC50 of 0.76 mM. Ibuprofen concentration-dependently decreased iNOS mRNA levels, with an IC50 > 2 mM. Thus, there was no correlation between decrease in iNOS activity and reduction in iNOS mRNA levels. Ibuprofen decreased iNOS protein levels, as determined by Western blot, with an IC50 of 0.89 mM. The data suggest that the reduction in iNOS activity by ibuprofen is due to inhibition of post transcriptional processing of this enzyme. Ibuprofen had no effect on constitutive COX (COX-1) or inducible COX (COX-2) mRNA expression. However, ibuprofen inhibited PGE2 formation with an IC50 of 0.86 mM. The anti-inflammatory actions of ibuprofen have been related to inhibition of COX and, subsequently, reducing prostaglandin formation. Since the potency of ibuprofen for inhibition of PGE2 formation and reduction in iNOS activity are similar, it is suggested that, at therapeutically effective doses, a decrease in iNOS activity may also occur in vivo. Therefore, reduction in iNOS protein levels in the brain may have a role in preserving the integrity of neurons in individuals susceptible to Alzheimer's disease. PMID- 9406925 TI - Retinal Muller glia secrete apolipoproteins E and J which are efficiently assembled into lipoprotein particles. AB - We have shown that apolipoprotein E (ApoE) is synthesized by Muller cells, the major glial cell of the rabbit retina, and secreted into the vitreous after which it is taken up by retinal ganglion cells and rapidly transported into the optic nerve [Amaratunga et al., J. Biol. Chem. 271 (1996) 5628-5632]. In this report we demonstrate that the ApoE secreted by Muller cells in vivo and in culture is efficiently assembled into lipoprotein particles. Apolipoprotein J (ApoJ) is also synthesized by these cells and assembled into lipoprotein particles. The lipoproteins are triglyceride-rich and contain cholesterol esters and free cholesterol. They are heterogeneous, with densities between 1.006 and 1.18 and diameters between 14 and 45 nm. We discuss the possible role of these lipoproteins in supplying the needs of neurons for lipids, especially long axonal projection neurons such as retinal ganglion cells, which are vulnerable to age related neurodegenerative diseases including Alzheimer's disease. PMID- 9406926 TI - Lysophosphatidic acid potentiates ACh receptor currents by G-protein-mediated activation of protein kinase C. AB - The effect of lysophosphatidic acid (lysoPA) on acetylcholine (ACh)-evoked currents was examined using normal and mutant Torpedo nicotinic ACh receptors expressed in Xenopus oocytes. LysoPA enhanced ACh-evoked currents in a washing time- and dose-dependent manner at concentrations of 0.1-3 microM, reaching a maximum of 210% 30 min after treatment, and instead, higher concentrations of lysoPA potentiated to a lesser extent or inhibited the currents. Dose-response curve to ACh was not affected by treatment with lysoPA. Current potentiation by lysoPA was fully inhibited by a broad G-protein inhibitor, guanosine-5'-O-(2 thiodiphosphate) (GDPbetaS), but not by a Gi/o-protein inhibitor, pertussis toxin (PTX). Additionally, the selective protein kinase C (PKC) inhibitor, GF109203X, blocked the potentiation, although the effect of lysoPA was not affected by the selective cAMP-dependent protein kinase (PKA) inhibitor, H-89, or mitogen activated protein kinase inhibitor, PD98059. LysoPA (3 microM) enhanced currents to 130% in Ca2+-free extracellular solution, and to 150% still in the mutant ACh receptors lacking PKC phosphorylation sites. The potentiation was also completely blocked by GF109203X. These results indicate that lysoPA potentiates ACh receptor currents by PTX-insensitive G-protein-mediated activation of Ca2+-dependent/ independent PKCs with subsequent phosphorylation of the receptors and by an unknown factor or process activated by PKC activation. PMID- 9406927 TI - Islet amyloid polypeptide and calcitonin gene-related peptide expression are upregulated in lumbar dorsal root ganglia after unilateral adjuvant-induced inflammation in the rat paw. AB - After unilateral adjuvant-induced inflammation, expression of neuropeptides believed to be involved in the inflammatory response, e.g. substance P and calcitonin gene-related peptide (CGRP), is upregulated in innervating sensory neurons. Islet amyloid polypeptide (IAPP) is structurally related to CGRP and constitutively expressed in sensory CGRP-containing neurons; the role of IAPP in sensory neurons is unknown. To examine whether IAPP could play a role in inflammation, IAPP expression in L5 dorsal root ganglion (DRG) and its distribution in the dorsal horn were investigated after unilateral adjuvant induced inflammation in the rat paw and compared with CGRP, using in situ hybridization and immunocytochemistry. At 12 h and day 3, but not day 21, the percentage of nerve cell profiles expressing IAPP and CGRP mRNA was greater in the ipsilateral L5 DRG; these changes paralleled the occurrence of edema around the tarsotibial joint and a slight limp. IAPP expression in individual nerve cell profiles was higher in the ipsilateral L5 DRG at 12 h, but not at days 3 and 21; the corresponding CGRP mRNA level was higher at days 3 and 21. At day 3, the higher expression of IAPP and CGRP on the ipsilateral side was accompanied by increased numbers of immunoreactive DRG neurons and fibers in the spinal cord dorsal horn. Largely, expression of IAPP and CGRP seems to be co-ordinately regulated by localized inflammation, although the rapid, but transient, upregulation in DRG neurons of IAPP mRNA expression and the slower, but sustained, upregulation of CGRP mRNA expression may indicate dissociated regulation of the peptides. Thus, IAPP could play a role in the initial phase of localized inflammation. PMID- 9406928 TI - Regulation of NMDA receptor subunit messenger RNA levels in the rat brain following acute and chronic exposure to antipsychotic drugs. AB - Based on anatomical and biochemical observations a role of glutamate in schizophrenia has been postulated. In the present work we have investigated the gene expression for two families of NMDA receptor subunits (NR-1 and NR-2) following acute and chronic treatment with typical (haloperidol) and atypical (clozapine) antipsychotic drug (APD) in rats. A single injection of the two drugs elicited a significant increase in the mRNA levels of NR-2B in the nucleus accumbens, whereas only haloperidol was able to elevate NR-2A and NR-2B in the hippocampus. Following a 21 day treatment, significant differences in the regulatory pattern of NMDA-R subunits were observed. Haloperidol increased their mRNA levels in striatum whereas clozapine, consistent with its relatively weaker influence on nigro-striatal dopamine function, did not change the expression of NR subunits in this region. Both APD's were able to decrease the expression of NR 2 subunits in the hypothalamus, but only clozapine was capable of reducing NR-2C in frontal cortex and accumbens. The regulation of NMDA-R subunits in specific brain regions may represent a novel and important mechanism through which APD's exert some of their effects on brain function. PMID- 9406929 TI - New isoforms of the chick glutamate receptor subunit GluR4: molecular cloning, regional expression and developmental analysis. AB - To identify chick GluR4 isoforms, we used PCR to amplify a C-terminal region that is the site of alternative splicing in rat. We report here the cloning of three novel chick GluR4 isoforms. GluR4c has a 113-bp insert in the C-terminus, is expressed in flip and flop isoforms, is most strongly expressed in the cerebellum, midbrain and forebrain, and appears from embryonic day (E) 2.5 through at least post-hatching day (P) 2, with a peak of expression at E17. GluR4d has a 184-bp segment inserted at the 4c splice site, occurs as flip and flop isoforms, is expressed most strongly in cerebellum, hindbrain and forebrain, and is present from E11 through P2, with peak expression at E17. GluR4s is a shortened form that lacks the nominal 4th transmembrane and flip/flop domains and shares a common C-terminal region with GluR4. GluR4s is expressed most strongly in the hindbrain and cerebellum and its expression increases from E11 through P2. Experiments on purified cerebellar cells show that glia express GluR4c and GluR4d at combined levels nearly twice that of GluR4 and that flip isoforms predominate. In contrast, granule cells express GluR4c and GluR4d at a level comparable to GluR4 and express GluR4s at a level less than half that in cerebellar glia. Thus, the independence of alternative splicing at the flip/flop and C-terminal splice sites allows seven alternatively spliced forms of GluR4 to exist in chick CNS. This structural diversity increases the potential for functional diversity in neuronal and glial GluRs incorporating GluR4. PMID- 9406930 TI - Localization of putative calcium-responsive regions in the rat BDNF gene. AB - Brain-derived neurotrophic factor (BDNF) has potent trophic and protective actions on CNS neurons, including mesencephalic dopaminergic neurons, ventral forebrain cholinergic neurons and spinal motor neurons. To evaluate the effects of calcium and other second messengers on BDNF gene transcription, C6 glioma cells were treated for 4 h with the calcium ionophore A23187, forskolin + isobutyl-methyl-xanthine (IBMX), or the phorbol ester, 12-O-tetradecanoyl-phorbol 13-acetate. Semi-quantitative RT-PCR analysis revealed that A23187 treatment increased BDNF transcripts containing the protein coding exon by 4.4-6.4-fold. Alternate BDNF transcripts were elevated to varying degrees after treatment with this ionophore and a subset of these transcripts was elevated following forskolin + IBMX treatment. When co-incubated with the RNA polymerase inhibitor, actinomycin D, A23187-induced increases were reduced or abolished, suggesting that calcium-mediated regulation of BDNF mRNA expression occurs at transcriptional as well as post-transcriptional levels. Transient transfection experiments employing reporter constructs containing serial 5' deletions of alternate BDNF promoters suggested that A23187-induced elevations in BDNF exon 1b, 1d and 1e containing transcripts are mediated by putative calcium-responsive regions flanking all three of these exons. PMID- 9406931 TI - Prostaglandin H synthase-2 and cytosolic phospholipase A2 in the hypoxic-ischemic brain: role in neuronal death or survival? AB - The breakdown of membrane phospholipids and subsequent arachidonic acid metabolism to prostanoids is a well-documented brain response to cerebral ischemia. To further elucidate the components of this signal transduction pathway, immunocytochemistry was used to determine the levels of two potentially important enzymes, cytosolic phospholipase A2 (cPLA2) and prostaglandin H synthase-2 (PGHS-2), in the immature rat brain following moderate unilateral hypoxic-ischemia (HI). The CA1 pyramidal cells of the hippocampus which undergo delayed neuronal death on the injured side following HI demonstrated a significant induction of PGHS-2 immunoreactivity 48 h post-insult. However, a consistent increase in PGHS-2 was also evident in the resistant dentate granule cells at an earlier time point. Although PGHS-2 is present in both susceptible and resistant cell populations following HI, the possibility remains that divergence further down-stream in the pathway is responsible for selective vulnerability. In contrast to the neuronal PGHS-2 expression, cPLA2 immunoreactivity appears to be of glial origin with increases in and around the CAI-2 pyramidal cell layer at the 72-168-h time points. These results suggest that prostanoids are likely to serve important roles in HI brain damage and repair in infant brain. PMID- 9406932 TI - AP-1 and NF-kappaB stimulated by carbachol in human neuroblastoma SH-SY5Y cells are differentially sensitive to inhibition by lithium. AB - In order to identify potential actions of lithium, the primary therapeutic agent for bipolar affective disorder, on processes regulating gene expression, its effects on two transcription factors, AP-1 and NF-kappaB, were measured in human neuroblastoma SH-SY5Y cells. The cholinergic agonist carbachol concentration dependently stimulated AP-1 (EC50 = 2 microM) and NF-kappaB (EC50 = 14 microM). Pretreatment for 24 h with a therapeutically relevant concentration of lithium (1 mM) substantially inhibited (30-35%) carbachol-stimulation AP-1 but not NF kappaB. Inhibition of carbachol-induced AP-1 was directly related to the concentration of lithium (1-20 mM). Besides being differentially sensitive to inhibition by lithium, activation of AP-1 and NF-kappaB demonstrated different carbachol EC50 concentrations, and carbachol-induced activation of AP-1, but not NF-kappaB, was inhibited by treating cells with Ni2+, which blocks receptor mediated calcium influx. These findings demonstrate that one mechanism by which lithium can influence the expression of specific genes is through the selective modulation of signaling processes which emanate from cholinergic receptor stimulation. PMID- 9406933 TI - Continuous administration of the glutamate uptake inhibitor L-trans-pyrrolidine 2,4-dicarboxylate produces striatal lesion. AB - This study examined the effects of chronic intrastriatal infusion of L-trans pyrrolidine-2,4-dicarboxylate (PDC), a selective competitive inhibitor of high affinity glutamate transport systems, via osmotic minipumps in rats. Injection of PDC at the rate of 25 nmol/h for 14 days caused striatal lesion. Histological evaluation on frontal striatal sections showed that the lesion was circumscribed to a circular area showing a dramatic neuronal loss accompanied by gliosis and representing 30% of the whole striatal surface at the level of the injection site. A total loss of neurons expressing glutamate decarboxylase (GAD67), enkephalin or substance P mRNA was observed on a similar circular area, suggesting degeneration of the two populations of striatal efferent neurons. In the whole striatum outside the region devoided of hybridization signal, a selective 27% decrease in enkephalin mRNA expression occurred, suggesting a higher sensitivity of enkephalin neurons versus substance P neurons to glutamate uptake-mediated alterations. Injection of PDC at the rate of 25 nmol/h for 3 days produced striatal lesion of similar extent. In contrast, PDC at the rate of 5 nmol/h did not produce neuronal damage when administered over 14 days. This study provides new in vivo evidence that defective glutamate transport is one of the critical conditions that may give rise to toxicity of an endogenous transmitter system in the striatum, and may underlie neuronal death in neurodegenerative diseases. PMID- 9406934 TI - Immunocytochemical identification of pinopsin in pineal glands of chicken and pigeon. AB - Pinopsin is a blue-sensitive photoreceptive molecule possibly involved in photic entrainment of the circadian pacemaker in the chicken pineal gland. To characterize pinopsin as a circadian photoreceptor, antibodies were raised against the C-terminal portion of pinopsin. As expected from the divergence of the amino acid sequence of this region, the resultant antibody cross-reacted with neither chicken rhodopsin nor red-sensitive cone pigment (chicken red). In Western blot analysis, the antibody stained a single band of 42-kDa protein in a detergent-extract of chicken pineal membranes, suggesting that pinopsin (calculated molecular weight, 38187) might be glycosylated and/or palmitoylated. Immunocytochemical examination of pineal sections of the chicken and the pigeon with this antibody revealed strong positive images for most of the membrane structures in the lumen of the follicles. This antibody also stained string- and bulb-shaped structures of the chicken parafollicular cells, the morphology of which resembles those of retinal photoreceptor cells. In contrast to the predominant distribution of pinopsin, a monoclonal antibody specific for chicken red stained a smaller number of membrane structures in the lumen of chicken pineal follicles. These results strongly suggest that the chicken pineal gland contains at least two types of photoreceptive molecules, pinopsin (major) and chicken red (minor). We show that the former molecule is localized in parafollicular pinealocytes and in the outer segments of pinealocytes that make contact with the follicular lumen. PMID- 9406935 TI - Differential expression of sodium channel genes in retinal ganglion cells. AB - Action potential electrogenesis in the axons of retinal ganglion cells is supported by voltage-gated sodium channels, and a tetrodotoxin (TTX)-inhibitable sodium conductance participates in anoxic injury of these axons within the optic nerve. However, the subtypes of sodium channels expressed in retinal ganglion cells have not been identified. In this study, we used reverse transcription polymerase chain reaction (RT-PCR) and restriction enzyme mapping, together with in situ hybridization, to examine the expression of transcripts for sodium channel alpha-subunits I, II, III, NaG, Na6, hNE/PN1 and SNS, and beta-subunits 1 and 2, in the retina of the adult rat. RT-PCR yielded high levels of amplification of I, II, III, Na6, beta1 and beta2 transcripts. In situ hybridization demonstrated the presence of all these mRNAs in the cell bodies of retinal ganglion cells. Retinal ganglion cells thus express multiple sodium channel mRNAs, suggesting that they deploy several different types of sodium channels. PMID- 9406936 TI - Experimental diffuse axonal injury induces enhanced neuronal C5a receptor mRNA expression in rats. AB - Several studies suggest the involvement of the complement system in the pathophysiology of traumatic brain injury (TBI). Since the intrathecal generation of anaphylatoxin C5a has been shown to mediate inflammatory effects within the central nervous system, we sought to characterize the cellular expression of the mRNA for the C5a receptor (C5aR, CD88) in brains of rats with experimental diffuse axonal injury (DAI) by in situ hybridization. Infiltrating leukocytes expressing C5aR mRNA were seen in meninges and lateral ventricles as early as 4 h after induction of DAI. The number of infiltrating C5aR-positive cells increased gradually up to 24 h after trauma. Within the brain parenchyma, up-regulation of C5aR mRNA expression was first seen in cerebellar Purkinje cells within 8 h. At 24 h after TBI, expression of C5aR mRNA was widespread bilaterally throughout the cortex and cerebellum, the cellular expression being restricted to pyramidal neurons and Purkinje cells. The intensity of C5aR transcript signals on neurons increased further up to 96 h after trauma. Ligand binding of C5a to its receptor on neurons might mediate previously unknown functions, thus possibly leading to neurotoxicity and secondary neuronal damage after TBI. PMID- 9406937 TI - Different effects of oxidative stress on activation of transcription factors in primary cultured rat neuronal and glial cells. AB - We compared the cytotoxic effects of oxidative stress on neuronal and glial cells in vitro by examining the cell viability and changes in DNA-binding activities of transcription factors, AP-1 and CREB, using Trypan blue exclusion and electrophoretic mobility shift assay (EMSA), respectively. Neurotoxin 6 hydroxydopamine (6-OHDA) and H2O2 reduced the viability of both types of cells in time- and concentration-dependent manner. Both neurotoxins dose-dependently decreased DNA-binding activities in neuronal cells. The results of cell viability assay suggested that these changes may reflect the reduction in neuronal cell viability. In contrast, both reagents increased DNA-binding activities in glial cells, although they decreased cell numbers. These results suggest that the effects of oxidative stress on transcription factors is different in neuronal and glial cells. We also examined the effect of brain-derived neurotrophic factor (BDNF) on 6-OHDA- or H2O2-induced changes in DNA-binding activities. In neuronal cells, pre-treatment with BDNF prevented the decrease in DNA-binding activities induced by 6-OHDA or H2O2. In glial cells, the effect of BDNF on oxidative stress induced changes in DNA-binding activities in the 6-OHDA-treated group were opposite to those in H2O2-treated group. Our results suggest that 6-OHDA and H2O2 may exert their cytotoxic mechanisms through different signal transduction systems. PMID- 9406938 TI - Dopamine receptor gene expression in an animal model of 'behavioral dependence' on ethanol. AB - The steady-state levels of messenger RNA (mRNA) of five cloned dopamine (D) receptors were measured in five brain regions in rats in a recently developed animal model of 'behavioral dependence' on ethanol. One group of rats was given the choice between ethanol and water over a 9-month period and developed 'behavioral dependence' on ethanol (group a). This group was compared with a group given the choice between ethanol and water for only 2 months (not yet behaviorally dependent, group b), a group forced to consume ethanol as sole fluid over a 9-month period (not behaviorally dependent, group c) and ethanol-naive control rats. All groups were sacrificed 1 month after ethanol withdrawal. The concentrations of mRNA of D3-receptors in the limbic forebrain (which included the nucleus accumbens) were significantly lowered in groups a and b, but unchanged in group c. D3 mRNA levels were reduced in the hippocampus of group b and unchanged in the cortex, amygdala and striatum. No significant changes in the mRNA concentrations of D1-, D2-, D4- or D5-receptors were seen in the five brain regions in any group. In conclusion, chronic consumption of ethanol under the 'free-choice condition', which may best induce the drug-rewarding effect, leads to specific changes in the D3-receptor gene expression which were not seen after forced ethanol administration. Changes in D3 mRNA levels were, however, not a specific correlate of 'behavioral dependence', as they were also detected in rats not yet 'behaviorally dependent' (group b). PMID- 9406939 TI - Induction of protein tyrosine phosphatase epsilon transcripts during NGF-induced neuronal differentiation of PC12D cells and during the development of the cerebellum. AB - We have investigated a possible role played by protein tyrosine phosphatase epsilon (PTPepsilon), which was recently cloned and predominantly expressed in brain, in neural differentiation and function. During neuronal cell differentiation of PC12D cells triggered by NGF or FGF, PTPepsilon transcripts were transiently induced at a time between the appearance of transcripts for immediate-early genes and for neuronal cell-specific markers. PTPepsilon was the only PTPase whose transcripts were induced during PC12D cell differentiation among over two dozen PTPase transcripts so far examined. Moreover, in situ hybridization revealed that PTPepsilon transcripts were detected in the neural tube of day 12 postcoitum embryo, and in the nervous system including brain, spinal cord, and ganglions in a ubiquitous manner in late gestational stages. In 4-day-old neonatal mice, the transcripts were widely distributed in the central nervous system where the strongest expression was detected in the hippocampus, cerebral cortex, and olfactory bulb. Interestingly, in day 7 and 16 neonatal brains, the strongest PTPepsilon gene expression was localized in the granular cells of cerebellum, which might indicate that PTPepsilon is involved in the differentiation of the granular cells. The biological significance of PTPepsilon in neuronal differentiation and brain functions is discussed. PMID- 9406940 TI - Type 2 interleukin-1 receptor mRNA is induced by kainic acid in the rat brain. AB - The in situ hybridization technique was used to examine the expression of type 2 interleukin-1 receptor (IL-1R2) mRNA in the rat brain following the systemic injection of kainic acid at a convulsive dose. The expression of IL-1R2 mRNA was not detected in any brain regions of the saline-injected control rats. 8 h after the systemic injection of kainic acid, weak expression of IL-1R2 mRNA was observed in the dentate gyrus and basolateral amygdaloid nucleus. At 12 and 24 h after the injection of kainic acid, IL-1R2 mRNA was markedly induced in various brain regions including the CA1 and CA3 fields of the hippocampus, dentate gyrus, basolateral amygdaloid nucleus, piniform cortex, claustrum, tenia tecta, arcuate hypothalamic nucleus, dorsomedial hypothalamic nucleus, suprachiasmatic nucleus, tuberal magnocellular nucleus and supramammillary nucleus. In these regions, the signals of IL-1R2 mRNA were observed on likely neuronal cells. Around the mediodorsal thalamic nucleus and the paraventricular thalamic nucleus, dispersed intense signals were observed on the non-neuronal cells. In addition, the expression of the mRNA on the venules was observed at 12 h. The strength of signals significantly decreased by 48 h after the injection. These findings revealed the spatiotemporal induction of IL-1R2 mRNA in the rat brain following the systemic administration of kainic acid, which has shown to cause neuronal degeneration, suggesting the pathological roles of IL-1R2 in the brain. PMID- 9406941 TI - Selective effects of partial and severe lesions of the serotonergic systems on Met-enkephalin and substance P neurons in rat basal ganglia. AB - The effects of partial (80%) vs. severe (> 95%) depletion of serotonin (5-HT) on peptide expression in basal ganglia were examined using immunocytochemical and in situ hybridization histochemical approaches. Topographical analysis of the changes in Met-enkephalin (Met-enk) and substance P (SP) levels were performed on the rat striatum, globus pallidus and substantia nigra 3 weeks after injecting 3 microl (partial lesion) or 6 microl (severe lesion) 5,7-dihydroxytryptamine (6.6 microg/microl) into the anterior raphe nuclei. Both kinds of lesion led to significant increases (39-42%) in Met-enk immunoreactivity in the striatum; a corresponding increase (21%) was detected in the globus pallidus only after severe 5-HT depletion. Only the severe lesion increased the SP immunoreactivity in the striatum (32%) and substantia nigra (26%). Neither striatal preproenkephalin nor preprotachykinin levels showed significant differences with the control values. These results suggest that the neuronal accumulation of Met enk or SP may be attributable to post-transcriptional events, such as a blockade of the peptide release, and that 5-HT may, thus, exert a facilitatory influence on the striatal output neurons. The results obtained after partial lesion indicate a preferential sensitivity of striatal Met-enk vs. SP containing terminals to the 5-HT denervation. These differences are illustrated in selective regional changes in peptide labeling. These data point to some balance exerted by the serotonergic and dopaminergic inputs on these neuronal populations. PMID- 9406942 TI - Changes in voltage-dependent calcium channel alpha1-subunit mRNA levels in the kindling model of epileptogenesis. AB - The establishment of a focus of epileptiform activity in the hippocampus of the rat, using the kindling paradigm, leads to enhanced voltage-dependent calcium conductance of CA1 pyramidal neurones (G.C. Faas, M. Vreugdenhil, W.J. Wadman, Calcium currents in pyramidal CA1 neurones in vitro after kindling epileptogenesis in the hippocampus of the rat, Neuroscience 75 (1996) 57-67; M. Vreugdenhil, W.J. Wadman, Kindling-induced long-lasting enhancement of calcium in hippocampal CA1 area of the rat: relation to calcium-dependent inactivation, Neuroscience 59 (1994) 105-114). Using semi-quantitative in situ hybridization techniques, we investigated whether these changes were associated with an altered expression of the genes that encode for the alpha1A-E-subunits of the voltage dependent calcium channels (VDCC). Kindling epileptogenesis was induced in rats that received an electrical tetanic stimulation of the Schaffer collateral/commissural fibre pathway in the hippocampus twice daily. Two groups of rats were studied before the appearance of generalized seizures, one group after at least 5 generalized seizures (fully kindled) and one group was investigated at long-term (28 days) after the last seizure. During the initial stages of epileptogenesis, the alpha1A-, alpha1D- and alpha1E-subunit mRNA levels were significantly increased in the different hippocampal subareas in comparison to the levels in control animals. In contrast, alpha1B-subunit gene expression decreased in the CA area and dentate gyrus. No significant change was observed in the alpha1C-I and alpha1C-II expression. At the fully kindled stage, the only significant change was an up-regulation of the alpha1B-subunit mRNA levels in the CA3 area, 24 h after the last seizure. No change in VDCC alpha1-subunit gene expression was found in animals investigated long-term after the establishment of the fully kindled state. Thus, the VDCC alpha1-subunit gene expression is altered in a subclass-specific manner during the early stages of kindling and may play a role in the establishment of a kindled focus, possibly caused by an alteration of the population of VDCCs involved in neurotransmitter release. The absence of long lasting changes suggests that the maintenance of a kindled focus is not due to persisting alterations in VDCC alpha1 mRNA levels. PMID- 9406943 TI - Activation of heme oxygenase and consequent carbon monoxide formation inhibits the release of arginine vasopressin from rat hypothalamic explants. Molecular linkage between heme catabolism and neuroendocrine function. AB - Heme oxygenase (HO)-catalyzed degradation of cellular heme moieties generates biliverdin and equimolar amounts of carbon monoxide (CO), which has been implicated as a possible modulator of neural function. Technical difficulties preclude direct measurements of CO within intact nervous tissues; hence, alternative procedures are needed to monitor the formation and possible biologic functions of this gas. In the present study rat hypothalamic explants were found to generate 114 +/- 5 or 127 +/- 11 pmol biliverdin/hypothalamus/1 h (n = 3) upon incubation with 1 or 10 microM hemin, respectively. Ten micromolar zinc protoporphyrin IX (Zn-PP-IX), a known inhibitor of HO, significantly decreased the degradation of 10 microM hemin from 127 +/- 11 to 26 +/- 11 pmol biliverdin/hypothalamus/1 h (n = 3; P < 0.01). Biliverdin was the principal product of HO-dependent heme degradation, as its possible conversion into bilirubin was precluded by hemin-dependent inhibition of biliverdin reductase. Basal or hemin-supplemented hypothalamic incubations were also shown to generate sizable amounts of propentdyopents (PDPs), reflecting HO-independent degradation pathways which do not liberate CO and cannot be inhibited by Zn-PP-IX. Plotting the ratio of biliverdin to PDPs thus provided an index of the efficiency with which hemin was degraded through biochemical pathways involving CO. Under the experimental conditions of our study, the biliverdin/PDPs ratio varied from 0 to 32 or 15%, depending on the absence or presence of 1 or 10 microM hemin respectively: this suggested that the formation of CO was most efficient at 1 microM hemin. Under these defined conditions, 1 microM hemin was also found to inhibit the release of arginine vasopressin (AVP) evoked by depolarizing solutions of KCl. A series of experiments showed that the effect of hemin was counteracted by Zn-PP-IX, and also by tin-mesoporphyrin IX, which is even more selective in inhibiting HO; it was also attenuated in the presence of the gaseous scavenger ferrous hemoglobin. Furthermore, the inhibition of AVP release could be reproduced by omitting hemin and by incubating hypothalami under CO, whereas treatment with biliverdin had no effect. This suggested that the release of AVP was suppressed by HO degradation of hemin, yielding CO as a modulator of hypothalamic function. These observations may be relevant to diseases characterized by inappropriate secretion of AVP and enzymatic disturbances affecting the synthesis of heme and the formation of CO through the HO pathway (e.g., acute intermittent porphyria or lead intoxication). PMID- 9406944 TI - Molecular characterization of antipeptide antibodies against the 5-HT1A receptor: evidence for state-dependent antibody binding. AB - Differential immunohistochemical labeling is often observed using different antibodies against the same protein. Two polyclonal antipeptide antibodies against the 5-HT1A receptor have been generated by our group. The S1A-170 (aa 170 186) and 258 (aa 258-274) are specific for sites in the second extracellular loop and third intracellular loop, respectively [E.C. Azmtia, I. Yu, H.M. Akbari, N. Kheck, P.M. Whitaker-Azmitia and D.R. Marshak, Antipeptide antibodies against the 5-HT1A receptor, J. Chem. Neuroanat., 5 (1992) 289-298]. Comparison of the labeling patterns of these two antibodies and other antipeptide antibodies against the 5-HT1A receptor revealed that although similar populations of cells were labeled, individual antibodies favor certain staining patterns. Immunocytochemistry and western blotting results of transfected cell lines and brain tissue revealed the following: (1) both the S1A-170 and S1A-258 are specific for the 5-HT1A receptor when used for immunocytochemistry in transfected HEK-293 and COS-1 cells; (2) when expressed in cultured cell lines, the 5-HT1A receptor is differentially glycosylated dependent on cell type, and the S1A-258 is specific for only certain species on immunoblots; and (3) the S1A-258 and L5B7 [M. Riad, S. El Mestikawy, D. Derge, H. Gozlan, and M. Hamon, Visualization and quantification of central 5-HT1A receptors with specific antibodies, Neurochem. Int., 4 (1991) 413-423] label common bands at 40 and 70 kDa on immunoblots of hippocampal proteins, but show opposite staining intensities. These results provide evidence for the immunocytochemical specificity of both the S1A-170 and S1A-258 and suggest that the discrepancies noted in immunohistochemistry may be due in part to different molecular conformations. PMID- 9406945 TI - Regional differences in expression of transcripts for Na+/Ca2+ exchanger isoforms in rat brain. AB - The Na+/Ca2+ exchanger has a primary role in maintaining intraneuronal Ca2+ homeostasis. There are three distinct Na+/Ca2+ exchanger isoforms cloned from rat brain, NCX1, NCX2 and NCX3, which are the products of three different genes. In the present study, isoform expression in different regions of rat brain was determined by using reverse transcription PCR (RT-PCR) and Northern analysis. RT PCR detected all three Na+/Ca2+ exchanger isoforms in each region studied (brainstem/spinal cord, cerebellum, cerebral cortex, striatum/septum and hippocampus). Northern analysis was performed to determine the steady-state mRNA levels of each isoform. NCX1 had two transcripts, 14 and 7 kb, and the 7-kb transcript was predominant in brainstem/spinal cord, cerebellum and hippocampus. NCX2 expression (4.8-kb transcript) was an order of magnitude higher than NCX1 or NCX3 expression in all the five areas except brainstem/spinal cord where the 4.8 kb transcript was nearly absent. The third isoform (NCX3) had two transcripts, one was 6 kb and the other was 4 kb. The 6-kb transcript was predominant in brainstem/spinal cord and cerebellum. The results suggest that Na+/Ca2+ exchanger isoforms are expressed ubiquitously in rat brain but that each isoform shows a unique distribution within the brain. The exchanger probably participates in the regulation of intracellular calcium homeostasis in a wide range of cell types within the brain. Furthermore, individual cells may contain more than one type of exchanger isoform with distinct subcellular distributions. PMID- 9406947 TI - mRNA for the m4 muscarinic receptor subtype is expressed in adult rat brain cholinergic neurons. AB - A number of pharmacological, anatomical, and immunological studies have previously addressed the subtype identity of the hippocampal muscarinic pre synaptic autoreceptor. A preponderance of findings indicate that it is of the M2 pharmacological type. Both the m2 and m4 molecular subtypes exhibit M2 pharmacology and there are few drugs that differentiate between these receptors. Pharmacological attempts at defining the hippocampal autoreceptor have yielded conflicting results. The basal forebrain is relatively enriched in m2 muscarinic receptor mRNA and protein, and lesions that denervate the hippocampus of its basal forebrain cholinergic input have shown a decrement in m2, but not m4, receptor protein in the hippocampus. Thus, the anatomical data obtained to date tend to support the view that the m2 subtype is expressed as the hippocampal autoreceptor. We have combined in situ hybridization histochemistry (ISHH) with immunocytochemistry to choline acetyltransferase to examine whether mRNA for the m4 subtype of muscarinic receptor is expressed in central cholinergic neurons. The m4 muscarinic mRNA was found at moderate levels in all subdivisions of the cholinergic basal forebrain, including the medial septum/diagonal band complex (MS/DB). The m4 mRNA was also found in striatal cholinergic interneurons, in the cholinergic reticular core of the upper brainstem, and in brainstem cholinergic motor neurons. Muscarinic m4 receptor mRNA was also found in many non-cholinergic cells in the brain. For example, the hippocampal pyramidal neurons, dentate gyrus granule cells, and entorhinal cortical pyramidal neurons express relatively high levels of m4 mRNA, while in the brainstem the dorsal raphe and pontine reticular nuclei express relatively high levels of this mRNA. The finding of m4 mRNA in the MS/DB cholinergic neurons suggests that this receptor protein might be expressed as an autoreceptor in hippocampal cholinergic terminals. PMID- 9406946 TI - Alterations in behavior and opioid gene expression induced by the novel tropane analog WF-31. AB - The effects of the acute administration of the serotonin-selective tropane analog, [2beta-propanoyl-3beta-(4-isopropylphenyl)-tropane, WF-31, on spontaneous locomotor activity were measured and compared to those of the highly selective serotonin uptake inhibitor, fluoxetine and cocaine, a non-selective re-uptake inhibitor of dopamine and serotonin. WF-31 (1, 10 and 30 mg/kg)-elicited increases in locomotor behaviors when compared to vehicle-treated rats. This increased activity was blocked by pre-treatment with the dopaminergic antagonist, flupenthixol, suggesting that these effects may be mediated by dopaminergic mechanisms. Cocaine, but not fluoxetine, also elicited increases in behaviors. In addition, the effects of these three compounds on opioid peptide gene expression were also assessed using in situ hybridization histochemistry in the same animals. The acute administration of both WF-31 and cocaine increased the expression of preprodynorphin mRNA in the dorsal striatum whereas fluoxetine had no effect. Expression of striatal preproenkephalin mRNA was augmented by all three compounds. Within the nucleus accumbens, PPD mRNA levels were affected only by treatment with WF-31, an effect that was blocked by pre-treatment with flupenthixol. In contrast, the acute administration of both WF-31 and fluoxetine, but not cocaine, increased the expression of preproenkephalin mRNA. These increases, however, were not reversed by pre-treatment with flupenthixol. Despite its profile in vitro as a relatively selective serotonin re-uptake inhibitor, some of the in vivo actions of WF-31 appear to be mediated by dopaminergic mechanisms. These data further suggest that the mechanisms underlying expression of the opioid peptides in the nucleus accumbens may vary from those in the dorsal striatum. PMID- 9406948 TI - Expression of tyrosine hydroxylase in magnocellular hypothalamic neurons of obese (fa / fa) and lean heterozygous (Fa / fa) Zucker rats. AB - In the magnocellular hypothalamic neurons (MCN) of normal rats, tyrosine hydroxylase (TH) is expressed in response to hyperosmotic stimulation and co exists with vasopressin. The present study shows that both Zucker obese (fa / fa) and heterozygous lean (Fa / fa) rats express TH in MCN independently of an osmotic challenge. The lack of L-DOPA and aromatic-L-aminoacid decarboxylase in the MCN showed the absence of mechanisms necessary for catecholamine synthesis in these cells. Therefore, TH in MCN seems to be functionally inactive and is not involved in catecholamine abnormalities observed in these rats. All TH immunoreactive MCN co-expressed vasopressin mRNA while a part of them co expressed oxytocin mRNA. This suggests a mechanism of regulation of TH expression in MCN which is different in Zucker rats and in dehydrated normal rats. PMID- 9406949 TI - Reversible sedimentation and masking of nerve growth factor (NGF) antigen by high molecular weight fractions from rat brain. AB - Nerve growth factor (NGF) was recently found to be largely associated with sedimentable fractions of adult rat brain and treatments of the fractions by alkaline pH increased the measurable amount of their NGF antigen as well as its solubilization [M.C. Hoener, E. Hewitt, J.M. Conner, J.W. Costello and S. Varon, Nerve growth factor (NGF) content in adult rat brain tissues is several-fold higher than generally reported and is largely associated with sedimentable fractions, Brain Res., 728 (1996) 47-56; M.C. Hoener and S. Varon, Effects of sodium chloride, Triton X-100, and alkaline pH on the measurable contents and sedimentability of the nerve growth factor (NGF) antigen in adult rat hippocampal tissue extracts, J. Neurosci. Res., in press (1997); C. Zettler, D.C.McL. Bridges, X.-F. Zhou and R.A. Rush, Detection of increased tissue concentrations of nerve growth factor with improved extraction procedure, J. Neurosci. Res., 46 (1996) 581-594]. We have further investigated the reversibility of these pH effects. Reversal of the pH of an adult rat hippocampal tissue extract from 10.5 to 7.4 led to an almost complete transfer of NGF back from nonsedimentable to sedimentable fractions and to a remasking of the previously unmasked portion of NGF antigen. Thus, molecules causing masking and sedimentation of NGF at pH 7.4 were likely to be present in the alkaline extract. A gel filtration column in PBS, pH 10.5 was used to separate such putative binding molecules from the NGF. All of the NGF antigen from rat hippocampal alkaline extract was found to elute with 19 kDa fractions. The same apparent molecular weight was found for mouse submaxillary beta-NGF and recombinant human beta-NGF. Masking and sedimentation no longer occurred when newly generated 19 kDa rat brain NGF was returned to pH 7.4. When high molecular weight fractions derived from the same gel filtration (in PBS, pH 10.5) were added back to the 19 kDa NGF pool at pH 7.4 and the mixture incubated and centrifuged, the measurability of 19 kDa rat brain NGF antigen was markedly reduced and half of the antigen was recovered in sedimentable fractions. Similar but less dramatic results were obtained when mixing the same high molecular weight fractions with 19 kDa mouse or human beta NGF. These findings provide new opportunities to identify molecules to which NGF may be bound within intact brain tissues. PMID- 9406950 TI - Distribution of N-acetylaspartylglutamate immunoreactivity in human brain and its alteration in neurodegenerative disease. AB - The dipeptide N-acetylaspartylglutamate (NAAG) may be involved in the process of glutamatergic signaling by both acting at glutamate receptors and as a glutamate protransmitter. In the present study we determined the cellular localization and distribution of NAAG-like immunoreactivity (NAAG-LI) in normal human brain and in neurodegenerative disorders to ascertain the degree of NAAG's colocalization to putative glutamatergic pathways. Immunohistochemistry with an antibody against NAAG was performed on control, Huntington's disease (HD) and Alzheimer's disease (AD) human autopsy and biopsy brain sections from the cerebral cortex, hippocampus, amygdala, neostriatum, brainstem and spinal cord. In normal human brain, NAAG-LI was widespread localized to putative glutamatergic pyramidal neurons of the cerebral cortex and hippocampus. Punctate NAAG-LI was present in areas known to receive neuronal glutamatergic input, such as layer IV of the cerebral cortex, striatal neuropil, and the outer portion of the molecular layer of the hippocampal dentate gyrus. In the two pathologic brain regions examined, the HD neostriatum and the AD temporal cortex, we observed a widespread loss of NAAG-LI neurons. In addition NAAG-LI reactive microglia surrounding plaques were seen in AD temporal cortex but not in the HD striatum. Our results suggest that NAAG is substantially localized to putative glutamatergic pathways in human brain and that NAAG-LI neurons are vulnerable to the neurodegenerative process in HD and AD. PMID- 9406951 TI - Reduced activity of the pyruvate dehydrogenase complex but not cytochrome c oxidase is associated with neuronal loss in the striatum following short-term forebrain ischemia. AB - Previous studies have identified changes in the activities of the pyruvate dehydrogenase complex (PDHC) and cytochrome c oxidase during early recirculation following short-term cerebral ischemia. However, the relationship of these changes to the delayed selective neuronal loss that develops as a result of short term ischemia is incompletely defined. The effects of ischemia and recirculation on the activities of these enzymes in the dorsolateral striatum, a region containing many susceptible neurons, and the ischemia-resistant paramedian cortex have been compared. No significant loss of activity of cytochrome c oxidase was seen in either region during the first few hours of recirculation following 30 min of ischemia. A decrease (of 32%) was observed at 24 h in the dorsolateral striatum. However, this probably resulted from changes in the mitochondrial fraction due to advanced neuronal degeneration. By contrast, there was a significant decrease (by 24%) in activity of PDHC at 3 h following a 30-min, but not a 10-min, ischemic period. Only the 30-min ischemic period resulted in extensive delayed neuronal loss. In the paramedian cortex, there was no significant change in PDHC and no neuronal loss following either ischemic period. These results provide strong evidence for a close association between neuronal loss and changes in the activity of PDHC but not cytochrome c oxidase in the dorsolateral striatum. PMID- 9406952 TI - Characterization of the extracellular serotonin release in the substantia nigra of the freely moving rat using microdialysis. AB - The characteristics of the serotonin release were investigated in the substantia nigra (SN) of the freely moving rat using microdialysis. We also examined whether the delay between surgery and microdialysis experiments might influence these characteristics by implanting rats with a guide cannula 1 or 2 days prior to microdialysis experiments. In the first group, the tissue was not punctured until the microdialysis probe was inserted the evening before the experiment. In the second group, the nigral tissue was punctured with an extended obturator which was then replaced by a microdialysis probe the evening before the experiment. After administration of 60 mM K+ a more pronounced increase in serotonin was observed in the first group (260%) compared to the second group (159%). Calcium free and tetrodotoxin (TTX, a sodium channel blocker) (1 microM) perfusion reduced extracellular serotonin to respectively 77% and 80% in the first group and 70% and 64% in the second group. These results suggest that vesicular release of nigral serotonin only occurs partially in this region and that minimizing the damage caused by implantation of the probe results only in 10% more vesicular release of serotonin. However, blockade of the serotonin reuptake carrier caused more TTX sensitivity of the serotonin release. Also, stimulation of the dorsal raphe by locally perfusing 60 mM K+ decreased serotonin in the SN, confirming the anatomical and functional link between both areas. PMID- 9406953 TI - Late-onset lipid peroxidation and neuronal cell death following transient forebrain ischemia in rat brain. AB - We previously reported that iron deposition was seen in the cerebral cortex and hippocampal CA1 area late after transient forebrain ischemia generated by four vessel occlusion in rats. Iron deposition in the hippocampal CA1 area was coupled with delayed pyramidal cell death, while that in the cerebral cortex was not accompanied by neuronal death or atrophy until 6 months after ischemia. Iron is involved in the formation of free radicals, thus contributing to lipid peroxidation. To elucidate whether this iron has deleterious effects on neurons, we investigated changes in the levels of lipid peroxidation and resulting neuronal damage in this ischemia model. The level of malondialdehyde plus 4 hydroxynonenal as major decomposition products of lipid peroxidation, monitored for 6 months beginning just after 30 min of transient forebrain ischemia, was significantly increased in the cerebral cortex at 6 months, and in the striatum from 1 week to 6 months compared to that in sham-operated controls. Histological changes were also examined up to 1 year after reperfusion by immunohistochemical methods. In contrast with the hippocampus and striatum, the cerebral cortex did not develop severe neuronal cell death and atrophy until 1 year after the ischemic insult. We showed that lipid peroxidation took place not only immediately after ischemia-reperfusion but also late after the ischemic insult in regions where iron was deposited, and we showed that neuronal cell death in the cerebral cortex appeared extremely late, suggesting that iron-mediated lipid peroxidation may be of importance in some slowly progressive forms of neurodegeneration. PMID- 9406954 TI - Glutamate metabotropic receptor agonist 1S,3R-ACPD induces internucleosomal DNA fragmentation and cell death in rat striatum. AB - Glutamate metabotropic receptor mediated mechanisms have been implicated in both neuroprotection and neurotoxicity. To characterize these mechanisms further in vivo, the effects of an intrastriatally injected metabotropic receptor agonist, trans-(1S,3R)-1-amino-1,3-cyclopentanedicarboxylic acid (1S,3R-ACPD), were studied alone and together with N-methyl-D-aspartate (NMDA) or kainic acid (KA) receptor agonists on DNA fragmentation and nerve cell death. 1S,3R-ACPD induced internucleosomal DNA fragmentation of striatal cells in a dose-dependent manner. TUNEL and propidium iodide staining showed DNA fragmentation and profound nuclear condensation around the injection site. Fragmented nuclei were occasionally seen under light microscopy. Internucleosomal DNA fragmentation induced by 1S,3R-ACPD was attenuated by the protein synthesis inhibitor cycloheximide as well as by the non-selective and selective metabotropic receptor antagonists L-(+)-2-amino-3 phosphonopionic acid (L-AP3), (RS)-aminoindan-1,5-dicarboxylic acid and (RS) alpha-methylserine-o-phosphate monophenyl ester, respectively. The 1S,3R-ACPD (100-900 nmol) induced death of striatal neurons was suggested by the reduction in NMDA and D1 dopamine receptors by up to 13% (P < 0.05) and 20% (P < 0.05) as well as by the decline in GAD67 mRNA (25%, P < 0.01) and proenkephalin mRNA levels (35%, P < 0.01). Interestingly, 1S,3R-ACPD attenuated internucleosomal DNA fragmentation induced by NMDA, but potentiated that induced by KA. These results suggest that metabotropic receptor stimulation leads to the death of striatal neurons by a mechanism having the biochemical stigmata of apoptosis. Moreover, metabotropic receptor stimulation evidently exerts opposite effects on pre- or postsynaptic mechanisms contributing to the NMDA and KA-induced apoptotic-like death of these neurons. PMID- 9406955 TI - Neuroprotection by the novel calcium antagonist PCA50938, nimodipine and flunarizine, in gerbil global brain ischemia. AB - Calcium is involved in the physiopathology of cerebral ischemia. Calcium antagonists might prevent the calcium overload and death of cells from ischemically compromised tissue. We compare the neuroprotective effect of various doses (0.2, 0.5 and 1 mg/kg) of two dihydropyridines, nimodipine and the novel 1,4-dihydropyridine derivative PCA50938, and flunarizine in the gerbil model of global ischemia. Improvements in morbidity were observed 2 h after the end of carotid occlusion (McGraw's scale) with 0.5 mg/kg of flunarizine, all doses of PCA50938 and 0.2 mg/kg nimodipine. Neuronal loss in the CA1 sector of the hippocampus was examined. The animals treated with 0.5 mg/kg flunarizine and those treated with 1 mg/kg PCA50938 showed a significant reduction in the percentage of damaged neurons in the hippocampal CA1 area, 72 h after transient ischemia. None of the animals treated with 0.5 mg/kg flunarizine had more than 80% of the evaluated neurons altered. We conclude that PCA50938 and flunarizine may act as neuroprotective drugs with different patterns of dose-response and neuroprotective-morbidity-mortality relationships, in the model of global cerebral ischemia in the gerbil. Flunarizine has a narrow therapeutic range. PMID- 9406956 TI - Chronic exposure to inorganic lead increases high-threshold voltage-gated calcium currents in rat pheochromocytoma (PC12) cells. AB - Rat pheochromocytoma (PC12) cells were exposed to lead acetate (0, 10, 25 and 50 microM) in their growth media for up to 12 weeks. High-threshold voltage-gated calcium currents were recorded each week from nerve growth factor-differentiated PC12 cells using the whole-cell patch-clamp technique. Chronic exposure for 1 month did not modify peak or sustained calcium current amplitudes in lead-treated cells when compared to sister control cultures. Two month exposure to 25 and 50 microM significantly increased peak and sustained calcium current amplitudes, while 10 microM had little effect. During the third month of exposure, peak and sustained calcium current amplitudes remained increased in the cells exposed to 25 and 50 microM lead acetate. By the end of the second month of exposure to 25 and 50 microM lead acetate, the voltage at which maximal current amplitude was attained shifted from + 10 mV to 0 mV. The observed effects of toxicologically relevant lead concentrations on high-threshold calcium currents in chronically exposed mammalian cells provide further support for the notion that at least one cellular target of the heavy metal's neurotoxic action may be the voltage-gated calcium channel. PMID- 9406957 TI - The N-methyl-D-aspartate (NMDA) receptor is postsynaptic to substance P containing axon terminals in the rat superficial dorsal horn. AB - The N-methyl-D-aspartate (NMDA) receptor is thought to mediate the postsynaptic effects of excitatory amino acids released from primary afferent terminals in the superficial layers of the dorsal horn of the spinal cord where synergistic associations with substance P (SP) have been implicated in the production of hyperalgesia. We examined the electron microscopic dual immunocytochemical localization of SP and the R1 subunit of the NMDA receptor (NMDAR1) in this region to determine the cellular basis for interactions between SP and NMDA receptor ligands. Of 971 profiles immunolabeled for NMDAR1, 40% were dendrites and the remainder were primarily unmyelinated axons and astrocytic processes. In dendrites, NMDAR1-like immunoreactivity (NMDAR1-LI) was associated with synaptic and non-synaptic portions of the plasma membrane, as well as intracellular membranes including smooth endoplasmic reticulum. These NMDAR1-labeled dendrites received synaptic input from unlabeled terminals and from terminals containing SP and/or NMDAR1-LI and they occasionally (25/389) also contained SP. In contrast, of 540 SP-immunoreactive profiles, 60% were axon terminals and the majority (252/324) of these SP-labeled terminals were presynaptic to NMDAR1-containing dendrites. These results provide anatomical evidence that the synergistic nociceptive effects of SP and NMDA ligands are attributed mainly to dual modulation of the activity of single dendritic targets in the dorsal horn of the spinal cord. They also suggest that activation of NMDA receptors may also play a role in the modulation of SP neurons, presynaptic release of SP or other neurotransmitters, and in glial function in the dorsal horn. PMID- 9406958 TI - Capsaicin-induced sensitization of primate spinothalamic tract cells is prevented by a protein kinase C inhibitor. AB - Protein kinase C (PKC) has been shown to be involved in nociceptive transmission in the spinal cord. This study tested the hypothesis that induction of central sensitization in the dorsal horn by an intradermal capsaicin injection involves activation of PKC. A PKC inhibitor (NPC15437) was infused through a microdialysis fiber into the spinal cord prior to capsaicin injection. The responses of spinothalamic tract (STT) cells were recorded before and after infusion of NPC15437, and after injection of capsaicin. STT cells show an increased background activity and increased responses to innocuous stimuli following capsaicin injection while responses to heat are decreased. Spinal infusion of the PKC inhibitor, NPC15437, had no effect on background activity or responses to peripherally applied stimuli prior to capsaicin injection. However, NPC15437 prevented the sensitization of cells to weak mechanical stimuli (brush and pressure) that occurs following capsaicin injection. NPC15437 had no effect on the increased background activity or decreased responses to heat stimuli induced by capsaicin injection, suggesting alternative mechanisms for these responses. These data suggest that PKC is important for the development of central sensitization to peripheral mechanical stimuli. PMID- 9406959 TI - Behavioral interactions have rapid effects on immunoreactivity of prohormone and gonadotropin-releasing hormone peptide. AB - The nervous system responds to both internal and external cues, integrating these signals to coordinate behavior and physiology. Mating interactions can promote dramatic changes in neuroendocrine cells which trigger successful copulation, ovulation, fertilization, and pregnancy. The neurons that transduce behavioral cues into neuroendocrine signals are distributed in a loose continuum along the medial ventral forebrain where they produce and secrete gonadotropin-releasing hormone (GnRH). In the past we have reported changes in GnRH-immunoreactive (GnRH ir) cell numbers in brains of female musk shrews sacrificed during, and after, brief mating interactions. The purpose of the current study was twofold: first to determine which aspect of intracellular GnRH production is stimulated by behavioral interactions; second, to characterize the specific aspects of the social exchange that trigger GnRH production. We report that 1 h after copulation the production of proGnRH protein is stimulated. Non-copulatory behavioral interactions resulted in a rapid decrease in the numbers of neurons containing GnRH-ir peptide. This change was accompanied by an increase in the GnRH-ir fibers in the median eminence, but no surge in luteinizing hormone. These data suggest that behavioral interactions stimulate release of mature GnRH peptide from cell bodies followed by accumulation of available GnRH in cell terminals. Copulation triggers increased production of proGnRH in cell bodies. The data highlight the usefulness of behavioral paradigms for the examination of the dynamics of neuropeptide production. PMID- 9406960 TI - Evidence for 5-HT4 receptor involvement in the enhancement of acetylcholine release by p-chloroamphetamine in rat frontal cortex. AB - The roles of endogenous serotonin (5-HT) and 5-HT receptor subtypes in regulation of acetylcholine (ACh) release in frontal cortex of conscious rats were examined using a microdialysis technique. Systemic administration (1 and 3 mg/kg, i.p.) of the 5-HT-releasing agent p-chloroamphetamine (PCA) elevated ACh output in a dose dependent manner. Depletion of endogenous 5-HT by p-chlorophenylalanine significantly attenuated the facilitatory effect of PCA on ACh release. The PCA (3 mg/kg)-induced increase in ACh release was significantly inhibited by local application of the 5-HT4 receptor antagonists RS23597 (50 microM) and GR113803 (1 microM), while the 5-HT1A antagonist WAY-100135 (10 mg/kg, i.p.; 100 microM), 5 HT(1A/1B)/beta-adrenoceptor antagonists (-)-pindolol (8 mg/kg, i.p.) and (-) propranolol (150 microM), 5-HT(2A/2C) antagonist ritanserin (1 mg/kg, i.p.; 10 microM) and 5-HT3 antagonist ondansetron (1 mg/kg, i.p.; 10 microM) failed to significantly modify the effect of PCA. These results suggest that PCA-induced enhancement of 5-HT transmission facilitates ACh release from rat frontal cortex at least in part through 5-HT4 receptors. PMID- 9406961 TI - Orphanin FQ has an inhibitory effect on the guinea pig ileum and the mouse vas deferens. AB - The activity of the recently isolated endogenous opioid-like peptide orphanin FQ (OFQ) was measured in two classical bioassays of opioid action. OFQ potently and concentration-dependently suppressed the electrically stimulated contractions of the guinea pig ileum (GPI) and the mouse vas deferens (MVD) with EC50 values of 1.82 +/- 0.16 and 2.97 +/- 0.01 nM, and Emax values of 56 +/- 3% and 96 +/- 4%, respectively. This effect of OFQ, in both the GPI and MVD, was insensitive to the opioid receptor antagonist naloxone. OFQ competed with [3H]diprenorphine binding to mu-, delta- or kappa-opioid receptors stably expressed in Chinese hamster ovary cell lines with IC50 values of 2.1 +/- 0.4, 2.2 +/- 0.3, 0.75 +/- 0.3 microM, respectively. Low affinity for the classical opioid receptors together with the inability of naloxone to antagonize its effect suggest that the inhibitory action of OFQ is mediated via a distinct OFQ receptor in the GPI and MVD. Consequently, the MVD could serve as a valuable bioassay of potential OFQ receptor antagonists. PMID- 9406962 TI - Role of angiotensin II receptor subtypes in mediating the sympathoexcitatory effects of exogenous and endogenous angiotensin peptides in the rostral ventrolateral medulla of the rabbit. AB - The pressor region in the rostral part of the ventrolateral medulla (VLM) in the rabbit contains a high density of the AT1 subtype of angiotensin (Ang) II receptor. In this study in anaesthetized barodenervated rabbits, we determined the effect of microinjection into the rostral VLM of the AT1 receptor antagonist losartan and the AT2 receptor antagonist PD123319 on resting arterial pressure and renal sympathetic nerve activity, and on the cardiovascular responses normally evoked by exogenous Ang II or Ang III in this region. Losartan (1 nmol) abolished the pressor and sympathoexcitatory responses normally evoked by exogenous Ang II, but PD123319 (1 nmol) had little effect on these responses. Both losartan (0.1-10 nmol) and PD123319 (0.1-1 nmol) had little effect on the resting arterial pressure and renal sympathetic nerve activity, except for a transient sympathoexcitatory response at the higher doses. In confirmation of previous findings, however, microinjection of the non-selective Ang receptor antagonist [Sar1,Thr8]Ang II (80 pmol) significantly decreased resting arterial pressure and sympathetic nerve activity. These results suggest that the sympathoexcitatory effects evoked by exogenous Ang II and III in the rostral VLM are mediated by AT1 receptors, but that the tonic sympathoexcitation produced by endogenous Ang peptides in the rostral VLM of the rabbit are mediated by receptors other than AT1 or AT2 receptors. PMID- 9406963 TI - Differential 5-HT-mediated regulation of stress-induced activation of proopiomelanocortin (POMC) gene expression in the anterior and intermediate lobe of the pituitary in male rats. AB - The purpose of the present study was to examine the role of 5-hydroxytryptamine (5-HT) neurons in mediating the effects of stress on proopiomelanocortin (POMC) gene expression in the anterior and intermediate lobes of the pituitary gland. To this aim, the effects of 5-HT depletion induced by administration of the neurotoxin 5,7-dihydroxytryptamine (5,7-DHT; 200 microg/rat; i.c.v.; 7 days) were investigated on POMC mRNA levels in the anterior and intermediate lobe of control and restraint-stressed rats. Three hours after brief exposure to diethyl ether (2 min) followed by 60 min of restraint stress increased POMC mRNA levels in the anterior and intermediate lobe of the pituitary. 5,7-DHT neurotoxic lesion, which resulted in a marked depletion of 5-HT (below the level of sensitivity of the neurochemical assay, 6 pg/sample) but not of dopamine or norepinephrine concentrations in the periventricular nucleus of the hypothalamus, had no effect on basal POMC mRNA levels in the anterior or intermediate lobe of the pituitary. However, 5-HT depletion further increased POMC mRNA levels in the anterior pituitary and completely blocked POMC mRNA level enhancement induced in the intermediate lobe of stressed rats. These results suggest a possible inhibitory 5 HT tone on POMC gene expression in the anterior pituitary and a stimulatory 5-HT tone in the intermediate lobe of the pituitary under these experimental conditions of stress. It appears, therefore, that 5-HT exerts a differential regulation of stress-induced activation of POMC gene expression in the anterior and intermediate lobes of the pituitary in male rats. PMID- 9406964 TI - Brain structures involved in the behavioral stimulant effect of central serotonin release. AB - Drugs such as p-chloroamphetamine or a combination of tranylcypromine and tryptophan release serotonin in the central nervous system and produce a behavioral serotonin syndrome. However, in the presence of methysergide or following destruction of descending spinal serotonergic projections by 5,7 dihydroxytryptamine, central serotonin release produces hyperlocomotion. This supports the hypothesis that release of serotonin in the brain promotes locomotion but that the expression of this effect can be blocked by concomitant intraspinal effects of serotonin release. Hyperlocomotion induced by serotonin release is attenuated or blocked by: (a) pretreatment with p-chlorophenylalanine; (b) acute surgical lesions of the basal diencephalon; (c) chronic lesions of the ventromedial midbrain tegmentum by local injection of 5,7-dihydroxytryptamine; and (d) acute surgical decortication. Medial decortication tends to be more effective then lateral decortication. Hyperlocomotion produced by methamphetamine is also attenuated or blocked by acute basal diencephalic lesions or decortication. It is suggested that ascending serotonergic and dopaminergic projections collaborate in the generation of spontaneous voluntary motor activity. PMID- 9406965 TI - Responses of primary afferents and spinal dorsal horn neurons to thermal and mechanical stimuli before and during zymosan-induced inflammation of the rat hindpaw. AB - Intraplantar administration of zymosan produces inflammation and results in behavioral evidence of hyperalgesia to mechanical and thermal stimuli in the rat. In the present studies, responses of primary afferents and spinal dorsal horn neurons to mechanical and thermal stimuli were examined before and during zymosan induced inflammation of the hindpaw. In tests of responses of primary afferents to mechanical stimuli, group mean mechanical response thresholds of C mechanonociceptor (CMN) units significantly decreased after zymosan administration. The group mean mechanical response thresholds of low threshold mechanoreceptor (LTM) units, A-mechanoheat (AMH) units, high threshold mechanoreceptor (HTM) units, and C-mechanoheat (CMH) units showed either no change or were increased significantly by intraplantar administration of zymosan. The group mean total discharges evoked during the 10 s mechanical stimulus were significantly increased after zymosan administration in CMN units. The group mean total discharges were either significantly decreased or unchanged in LTM, AMH, HTM, and CMH units. In tests of responses of spinal dorsal horn neurons to mechanical stimuli, the group mean mechanical response threshold of nociceptive specific (NS) units decreased significantly 1 h following administration of zymosan, whereas no significant changes occurred in the mechanical response thresholds of wide dynamic range (WDR) neurons in zymosan-injected rats, WDR neurons in saline-injected rats, or NS neurons in saline-injected rats. The group mean total discharges of only NS neurons were significantly increased during the 10 s mechanical stimulus 3 and 4 h after zymosan administration. In tests of responses of primary afferents to thermal stimuli, intraplantar administration of zymosan resulted in significant decreases in group mean response thresholds of CMH units and significant increases in group mean response thresholds of AMH units. The group mean total discharges of CMH units was either unchanged or significantly increased during thermal stimuli depending on both the time of testing and the temperature of the test stimulus. The group mean total number of discharges of AMH units was significantly decreased during tests of all thermal stimuli. In tests of responses of spinal dorsal horn neurons to thermal stimuli, intraplantar administration of zymosan resulted in significant decreases in thermal response thresholds of both WDR and NS units of zymosan-injected rats, but no changes in WDR and NS units of saline-injected rats. The group mean total discharges evoked by the 15 s thermal stimuli also increased significantly in both WDR and NS units after zymosan administration. Zymosan administration resulted in increased background activity only in CMH units. These increases occurred immediately following the injection and dissipated by the first hourly test period. Significant changes in background discharges of both WDR and NS units occurred at some hourly test intervals following administration of zymosan, but these changes were not consistent with respect to either unit type or modality of the test stimulus. These data suggest that the zymosan-induced hyperalgesia to mechanical stimuli observed in behavioral studies reflects decreases in response thresholds of peripheral CMN units and spinal NS neurons. Hyperalgesia to thermal stimuli reflects decreases in response thresholds of peripheral CMH units, spinal WDR neurons, and spinal NS neurons. These data support the view that different physiological substrates mediate hyperalgesia to either thermal or mechanical stimuli following intraplantar administration of zymosan. PMID- 9406966 TI - Immunohistochemical and RT-PCR detection of Na+-dependent inorganic phosphate cotransporter (NaPi-2) in rat brain. AB - Expression of a renal Na+-dependent inorganic phosphate (Pi) cotransporter (NaPi 2) was studied in rat forebrain with reverse transcription and polymerase chain reaction (RT-PCR) and immunohistochemistry. RT-PCR analysis for total RNA from whole brain and sequencing of the PCR products showed expression of NaPi-2 mRNA in the brain. Immunohistochemical analysis revealed NaPi-2 staining in many nonpyramidal neurons of all six layers throughout neocortical areas and in neurons of proisocortical and periallocortical areas. NaPi-2-immunoreactive neurons were also detectable in the piriform cortex, hippocampal formation, caudate-putamen, amygdaloid nuclei and lateral geniculate nucleus. Furthermore, NaPi-2 staining was shown in ependymal cells and microvascular endothelial cells. The present results suggest that NaPi-2 is synthesized within the brain and involved in maintaining Pi homeostasis of certain neurons and/or the entire brain. PMID- 9406967 TI - Carbonyl histochemistry in rat reperfusion nerve injury. AB - Free radical mediated, site-specific lipid and protein oxidation has been implicated in the pathophysiology of an ischaemic/reperfusion injury. The aim of the present study was to determine whether carbonyl formation could be detected histochemically in reperfused rat sciatic nerves. We also examined the effects of preischaemic alpha-tocopherol supplementation on carbonyl formation in reperfused nerves. Seven hours of near-complete ischaemia was induced in rat right hindlimb by occlusion of major arteries using microvascular clips. Histochemical detection of carbonyl compounds, applying naphthoic acid hydrazide (NAH) and Fast Blue B (FBB), was undertaken at thigh, knee and calf levels of sciatic, tibial and peroneal nerves. NAH-FBB reactivity was confined to vessels in reperfused nerves. Positively stained epi-, peri- and endoneurial vessels were invariably observed after 2 h of reperfusion at all levels examined. After 24 and 48 h and 7 days of reperfusion, NAH-FBB-positive vessels were more frequently found at knee and calf levels than at the thigh level. Following preischaemic alpha-tocopherol supplementation, no vessels were stained positively with NAH-FBB, except for some epineurial vessels at knee and calf levels after 2 h of reperfusion. Morphometry in endoneurial vessels at the knee level revealed that endothelial cell area in alpha-tocopherol-treated reperfused nerves was significantly less when compared with those in reperfused nerves without alpha-tocopherol. In conclusion, we have demonstrated histochemical evidence of carbonyl formation in vessels, but not with nerve fibres, in ischaemic/reperfused rat sciatic nerves. These abnormalities were prevented with preischaemic supplementation of alpha tocopherol. PMID- 9406968 TI - Upregulation of the expression of vasopressin gene in the paraventricular and supraoptic nuclei of the lithium-induced diabetes insipidus rat. AB - The expression of arginine vasopressin (AVP) gene in the paraventricular (PVN) and supraoptic nuclei (SON) was investigated in rats with lithium (Li)-induced polyuria, using in situ hybridization histochemistry and radioimmunoassay. The male Wistar rats consuming a diet that contained LiCl (60 mmol/kg) for 4 weeks developed marked polyuria. The Li-treated rats produced a large volume of hypotonic urine with low ionic concentrations. Plasma sodium concentrations were found to be slightly increased in the Li-treated rats compared with those in controls. Plasma concentration of AVP and transcripts of AVP gene in the PVN and SON were significantly increased in the Li-treated rats compared with controls. These results suggest that dehydration and/or the activation of visceral afferent inputs may contribute to the elevation of plasma AVP and the upregulation of AVP gene expression in the PVN and the SON of the Li-induced diabetes insipidus rat. PMID- 9406969 TI - Role of dorsal vagal motor nucleus in angiotensin II-mediated tachycardia in the conscious trout Oncorhynchus mykiss. AB - Responses of heart rate (HR) and mean arterial blood pressure (MABP) were examined following microinjection of angiotensin II ([Asn1,Val5]AI) within the dorsal vagal motor nucleus (DVN) of the conscious trout's brainstem. AII (15-125 fmol) preferentially and significantly increased HR in a dose-dependent manner, but the rise in MABP was not dose-dependent and was only significant (P < 0.05) after injection of AII at a dose of 62.5 fmol. The cardiovascular action of AII was site-specific, since administrations of the peptide at a dose of 62.5 fmol, but outside the boundaries of the DVN, were devoid of any effect on HR or MABP. All the responses to DVN injections of AII were totally prevented by DVN injection of 1 nmol of losartan, a mammalian non-peptide AII subtype 1 (AT1) receptor antagonist. The ability of DVN injection of AII to induce a tachycardic response was negatively correlated to HR basal values. In conclusion, these results indicate that, at femtomolar doses, AII exerts a central neurocardioregulatory role, involving a localized receptor closely related to the mammalian AT1 receptor subtype within the DVN of the trout. PMID- 9406970 TI - Tetrodotoxin blocks NPY-induced but not muscimol-induced phase advances of wheel running activity in Syrian hamsters. AB - During the middle of the subjective day, circadian activity rhythms in Syrian hamsters can be phase advanced by a variety of stimuli including microinjection of neuropeptide Y (NPY) or muscimol into the suprachiasmatic nucleus (SCN). It is not known, however, if these treatments shift activity rhythms by acting directly on pacemaker cells within the SCN. In the present study NPY and muscimol were microinjected with either tetrodotoxin or saline in order to determine whether classical synaptic transmission within the SCN is necessary for the phase advances produced by NPY or muscimol. Blockade of sodium-dependent action potentials within the SCN prevented NPY- but not muscimol-induced phase advances. These data, along with our previous finding that bicuculline blocks NPY-induced phase advances, suggest that NPY requires sodium-dependent action potentials within GABAergic neurons in order to phase-shift the circadian pacemaker. PMID- 9406971 TI - Modulation of anxiety-related behaviours following lesions of the prelimbic or infralimbic cortex in the rat. AB - A series of experiments examined behavioural and autonomic aspects of stress and anxiety in rats subjected to either: (1) electrolytic lesions of the infralimbic cortex subregion of the medial prefrontal cortex; (2) electrolytic lesions of the prelimbic cortex subregion of the medial prefrontal cortex; (3) sham lesions of infralimbic or prelimbic cortex (sham control); or (4) no lesions (control). In exploration-based models of anxiety, infralimbic- or prelimbic-lesioned rats spent less time in the centre of an open field and less time on the exposed arms of an elevated plus maze, indicating increased anxiety. Locomotor activity was normal in the lesioned rats when tested in a non-stressful enclosed environment. In a step-down passive avoidance task, infralimbic-lesioned rats stepped down more quickly than controls onto a grid floor where they had been shocked 24 h previously. Prelimbic-lesioned rats were no different to controls on this test, although they showed greater latencies to step down onto the grid floor during conditioning. In a final experiment, indirect calorimetry was used to show that both infralimbic- and prelimbic-lesioned rats have essentially normal alterations in oxygen consumption and energy substrate utilisation when exposed to brief footshock. Thus, the impaired passive avoidance in infralimbic-lesioned rats cannot be attributed to decreased nociception. It is concluded that both the prelimbic and infralimbic regions play a role in anxiety, and that this role may be subtly differentiated. In particular, the infralimbic cortex may have a specific role in mediating the inhibition of behaviours associated with aversive outcomes. PMID- 9406972 TI - Spermine does not compete with omega-conotoxin GVIA in the striatum radiatum of the hippocampal slice. AB - The effect of spermine (Spm) and of omega-conotoxin GVIA (CTX) on the population excitatory postsynaptic potentials (pEPSP) in stratum radiatum of the CA1 area were compared. CTX decreased irreversibly the initial slope of pEPSP by 57%. Spm produced a maximum inhibition of 85% with an apparent dissociation constant of 0.85 mM and a maximum Hill coefficient larger than 3. The effect of Spm was mostly reversible. Preincubation with Spm did not protect the slice from the irreversible effect of CTX suggesting that they interact with different sites. Since CTX and Spm inhibited pEPSPs with very different affinities and reversibilities a kinetic model was developed to compare their effects. This model relates the inhibitors' binding to presynaptic voltage-activated Ca2+ channels (VACC) with inhibition of pEPSP. The model suggest that: all CTX and Spm effects can be explained by inhibition of VACC. Spm and CTX do not compete for the same site. CTX inhibits 20% (N-type) and Spm 40% of channels (probably the Q type). More than three Spm molecules bind per one channel molecule, while one CTX is sufficient to inhibit channel function. The model also illustrates that the inhibitor concentration-pEPSP inhibition curves display a Hill coefficient similar to that for inhibitor binding. PMID- 9406974 TI - Differential effects of restraint stress on hippocampal 5-HT metabolism and extracellular levels of 5-HT in streptozotocin-diabetic rats. AB - Streptozotocin (STZ)-elicited diabetes reduces central serotonin (5 hydroxytryptamine, 5-HT) synthesis/metabolism, but whether this reduction leads to decreased release of 5-HT has only scarcely been investigated. We have thus analysed the impact of STZ diabetes on hippocampal extracellular 5-HT levels both under basal conditions and during restraint stress, a procedure known to stimulate hippocampal 5-HT synthesis/metabolism and release. The pretreatment with STZ (3 weeks beforehand) and the 1 h restraint session respectively decreased and increased hippocampal 5-HT metabolism, as assessed by tissue analysis of 5-HT and 5-hydroxyindoleacetic acid. On the other hand, hippocampal microdialysis revealed no difference in basal levels of extracellular 5-HT levels in (conscious) vehicle- and STZ-pretreated rats, but a differential effect of restraint. Thus, extracellular 5-HT levels increased throughout restraint (maximal increase: 194%) in vehicle-, but not in STZ-pretreated rats. In the latter rat group, plasma corticosterone levels were, however, increased, thus indicating a significant aversiveness to stress. Lastly, because anxiety-related behaviours may be affected by hippocampal serotonergic systems, resting and restrained vehicle- and STZ-pretreated rats were compared (immediately after stress) in an elevated plus-maze of anxiety. Pretreatment with STZ reduced the percent number of open arm entries and the number of closed arm entries, indicating increased anxiety and reduced locomotor activity, respectively. Restraint tended to increase anxiety-related behaviours in all rats, but this trend never reached significance. Our results confirm that gross analyses of 5-HT metabolism do not yield information on 5-HT release, and suggest that the prevalence of diabetes among patients suffering affective disorders could be related to the lack of hippocampal serotonergic response to aversive stimuli. PMID- 9406973 TI - Peripheral-type benzodiazepine receptor ligands and serum steroid hormones. AB - The peripheral-type benzodiazepine receptors (PBR) are involved in various cellular functions, including steroidogenesis. The impact of these receptor ligands has been demonstrated mainly in steroidogenic cells. The aim of the present study was to assess in intact female rats the effect of chronic (21 days) administration of the PBR ligands PK 11195 (15 mg/kg) and Ro 5-4864 (5 mg/kg), the mixed ligand diazepam (5 mg/kg), and the central benzodiazepine receptor ligand clonazepam (1 mg/kg) on PBR binding characteristics in steroidogenic (ovary and adrenal) and non-steroidogenic (uterus and kidney) organs, as well as on serum hormonal steroids (estradiol, progesterone, and corticosterone). Selective and mixed PBR ligands up-regulated PBR density in the two steroidogenic organs, while Ro 5-4864 also induced elevation of the receptor density in the non steroidogenic organs. In contrast to Ro 5-4864, PK 11195 treatment down-regulated renal PBR. Clonazepam elevated adrenal PBR. On the serum hormonal level, Ro 5 4864 suppressed estradiol secretion. The other ligands did not affect hormonal steroid levels. It appears that in female rats, at least at these doses and dosing schedules, there is no correlation between the impact of chronic in vivo exposure to these agents on PBR density and ovarian and adrenal hormone levels. PMID- 9406975 TI - GABA-immunoreactive neurones and interactions of GABA with serotonin and FMRFamide in a peripheral sensory ganglion of the pond snail Lymnaea stagnalis. AB - The osphradium is a putative chemosensory organ of aquatic molluscs. Previously, we identified cells with serotonin (5-HT) and FMRFamide (FMRFa)-like immunoreactivity in the osphradial ganglion of Lymnaea stagnalis. The present investigation has established the presence of cells immunoreactive to gamma aminobutyric acid (GABA). Some of these cells send processes to the sensory epithelium and are thus considered to be primary sensory neurones. Colocalisation of GABA and FMRFamide-like immunoreactivities was found in some of these and other neurones. The responses of the osphradial output electrical activity to the single and combined application of the above neuroactive substances were examined. 5-HT slightly increased and FMRFa decreased the activity. GABA alone was generally ineffective; however, it had a consistent stimulating effect after pretreatment with 5-HT. In its turn, pretreatment with GABA significantly increased the inhibitory action of FMRFa. Primary sensory neurones giving this kind of responses in the nerve were identified electrophysiologically and morphologically in the osphradial ganglion. Our results indicate that GABA takes part in relay of sensory signals into the central nervous system, and transmitter interactions involving GABA, 5-HT, and FMRFa are considerable for the final output pattern of the osphradial sensory network. PMID- 9406976 TI - Presynaptic action of the neurosteroid pregnenolone sulfate on inhibitory transmitter release in cultured hippocampal neurons. AB - The effects of the neurosteroid pregnenolone sulfate (PS) were studied in 3- to 9 week-old hippocampal cultures from neonatal rats. Cells were voltage clamped using CsCl filled electrodes, while action potentials and excitatory glutamatergic currents were abolished by superfusing with a combination of tetrodotoxin, 6-cyano-7-nitroquinoxaline (CNQX) and 2-amino-5-phosphonopentanoic acid (AP-5). Under these conditions spontaneous GABAergic inhibitory postsynaptic currents (sIPSCs) were seen as inward currents at a holding potential of -70 mV. Their amplitude distributions were skewed without clearly detectable peaks. PS at 1-50 microM concentrations decreased the frequency of sIPSCs, with 1 microM being the most effective concentration. The effect appeared after 10-15 min of steroid application and the magnitude of the reduction increased during the early wash period. No recovery of sIPSC frequency was found after 30 min of washing with steroid-free medium. sIPSC amplitudes were not significantly changed at the time the effect of PS on sIPSC frequency was observed. The slow onset of this effect and its duration suggest a novel presynaptic action of the neurosteroid PS on GABAergic inhibition in the mammalian brain. PMID- 9406977 TI - Removal of tissue plasminogen activator does not protect against neuronal degeneration in the cerebellum of the weaver mouse. AB - Tissue plasminogen activator (tPA) is a serine protease that has been shown to be involved in neuronal degeneration. Recently, elevated cerebellar tPA has been reported in a naturally occurring mutant mouse, weaver. Weaver mice suffer extensive degeneration of cerebellar granular neurons during development, leading to severe malformation of the cerebellum as well as abnormal behavior (ataxia). The observations that the developing weaver cerebellum displays a 10-fold increase in tPA activity over wild-type and that a serine protease inhibitor was able to rescue weaver granule cells from premature death in culture suggested that tPA might mediate the death of these mutant neurons. We tested this possibility by introducing the weaver mutation into tPA-deficient mice and comparing the weaver phenotype in the presence or absence of tPA. Analysis at 28 days after birth indicates that tPA-deficient weaver mice are indistinguishable from tPA-containing weaver mice in behavior, cerebellar anatomy, histology, and laminin expression (also reported to be increased in weaver). These results suggest that removal of tPA activity from weaver mice does not protect against neuronal degeneration in the cerebellum and, thus, tPA does not appear to mediate this form of cell death. PMID- 9406978 TI - Opponent effects of quinine and sucrose on single fiber taste responses of the chorda tympani nerve. AB - Responses of single chorda tympani fibers to mixtures of taste stimuli were studied in the golden hamster (Mesocricetus auratus). Sucrose-best neurons showed significant suppression to quinine-sucrose mixtures compared to sucrose alone. Quinine may exert its effect as an opponent stimulus in the receptor cells at the second messenger level. This suppression may make bitter quinine more readily detected when embedded in mixtures with sweeteners. PMID- 9406979 TI - Insulin-like growth factor-I receptors in normal appearing white matter and chronic plaques in multiple sclerosis. AB - Preclinical studies suggest that insulin-like growth factor-I (IGF-I) plays an important role in oligodendrocyte survival and myelination. We used human recombinant [125I]IGF-I to study IGF-I receptors in post-mortem brain tissue from patients with multiple sclerosis (MS). In normal appearing white matter, we found that IGF-I receptor densities and binding characteristics were not different between MS patients and controls. In chronic plaques, histologically characterized by astrogliosis, we found densities of IGF-I receptors which were in the same range as those measured in the normal appearing white matter. In vitro studies have shown that IGF-I also acts as a mitogenic factor for astrocytes. Since MS lesions are rapidly invaded by reactive astrocytes, IGF-I may not only protect oligodendrocytes and stimulate remyelination but also enhance the astrogliosis that limits repair. PMID- 9406980 TI - Hypoglycemia enhances the expression of mRNA encoding beta-amyloid precursor protein in rat primary cortical astroglial cells. AB - Deposition of beta-amyloid (A beta) is a characteristic feature of the pathology of Alzheimer's disease (AD). Since glucose metabolism and the consequential ATP production are depressed in the temporal and parietal regions of the cortex in patients with AD, we designed the present study to investigate the possible role of hypometabolism in the pathogenesis of AD. We incubated rat primary cortical astroglial cells for 2 h to 4 days in a media deprived of 95% of its glucose and assessed the expression and alternative splicing of the mRNA that encoding beta amyloid precursor protein (APP) using RT-PCR. Hypoglycemia caused a time dependent increase in APP mRNA expression, which reaches a peak level of 173.2% of control expression (P < 0.05) at 24 h of hypoglycemia. Noteworthy, hypoglycemia favors the alternative splicing that includes the exon 7 segment, which encodes a Kunitz-type serine protease inhibitor domain. This study demonstrates that hypoglycemia increases APP mRNA expression in astroglial cells and processing of APP mRNA to a form that may encourage A beta deposits in AD. These data suggest that the observed hypometabolism in AD may contribute to its deposition of A beta in affected brain regions. PMID- 9406981 TI - The effect of pinealectomy on the crypts of defunctioned rat colon. AB - Previously it has been found that 6 months after pinealectomy hyperplasia occurred in the crypt cells of the rat small bowel and colon. It is also known that defunctioning a loop of colon, using a colostomy, results in crypt cell hypoplasia, emphasizing the prime importance of luminal factors in the control of crypt cell proliferation. To determine if the effects of pinealectomy on the colon could be modified by the absence of colonic luminal contents, the crypt cell kinetic effects of combined pinealectomy and defunctioning of a colonic loop by colostomy for 6 months were examined by using a stathmokinetic technique. It was found that the hypoproliferative effect of defunctioning a loop of colon was largely but not completely overridden by the hyperproliferative effect of pinealectomy. However, previously it has been found that in the rat small bowel, the hypoproliferative effects of defunctioning a loop were completely overridden by the effect of pinealectomy. This and other evidence suggests that the role of the pineal in the control of crypt cell proliferation in the colon may possibly be different from its role in the small bowel. There is other evidence of possible involvement of the pineal in carcinoma of the colon and it is possible that its role in the colon may be to prevent excessive mitotic activity, which is known to be present in the early stages of carcinoma. The Pineal gland may have a role in modulating the usual mechanisms of crypt cell mitotic control. PMID- 9406982 TI - Supplemental melatonin increases clonal lifespan in the protozoan Paramecium tetraurelia. AB - The hypothesis that melatonin supplementation can increase the lifespan of a single-celled organism was tested by the administration of melatonin to the ciliated protozoan Paramecium tetraurelia. Melatonin supplementation in dim red light at a dose of 0.043 mM (10 mg/L) of nutrient media (bacterized Cerophyl) per day, followed by incubation for 23 hr in darkness, increased the mean clonal lifespan of Paramecium tetraurelia in days by percentages ranging from 20.8% (P < 0.01, two-tailed t-test) to 24.2% (P < 0.01, ANOVA) over controls. Maximum clonal lifespan in days was also increased in melatonin-supplemented cells, from 14.8% to 24.0% over controls. Mean clonal lifespan in fissions was not significantly greater in melatonin-supplemented cells, with values ranging from 6.0% to 15.5% over controls. Maximum clonal lifespan in fissions did not differ appreciably, with values ranging from 1.0% to 9.1% over controls, except in the case of cells selected for rapid division rates, in which melatonin-supplemented cells (393 fissions) lived 20.9% longer than controls (325 fissions) in terms of cumulative cell doublings during the clonal lifespan. The finding that melatonin supplementation increased clonal lifespan in Paramecium tetraurelia, an aerobic, single-celled organism, suggests that the mechanism of melatonin's longevity promoting effects may be intracellular. PMID- 9406983 TI - The effect of atenolol, a beta1-adrenergic antagonist, on nocturnal plasma melatonin secretion: evidence for a dose-response relationship in humans. AB - Pineal beta1-adrenergic receptors are involved in the regulation of melatonin secretion. The involvement of beta1-adrenergic receptors has been demonstrated by the ability of acute administration of beta-antagonists to suppress the nocturnal rise of circulating melatonin and its urinary metabolite 6-sulphatoxymelatonin (aMT6s). The present study was undertaken to examine the relationship between increasing doses of atenolol and nocturnal plasma melatonin concentrations. Six healthy subjects participated in the study for a period of 5 weeks. Subjects were administered placebo, 12.5, 25, 37.5, and 50 mg doses of atenolol in a randomized single blind design. Each dose was separated by a 1 week washout period. Blood samples were collected at regular intervals from 19.00 hr to 06.00 hr. Repeated measures analysis of variance showed a dose-dependent decrease in plasma melatonin concentrations (P<0.01). A Student Newman-Keuls post hoc test indicated significant differences between placebo and all doses of atenolol (P<0.05). The results demonstrate a dose-dependent relationship between beta1-receptor blockade and suppression of nocturnal plasma melatonin in humans. PMID- 9406984 TI - Melatonin entrainment of the circadian N-acetyltransferase rhythm in the newborn rat pineal gland. AB - In 15-day-old control and vehicle-treated rats, the evening rise of the pineal N acetyltransferase occurred at the same time as in their mothers, whereas in 5-day old pups, the rise occurred by 2-3 hr earlier. Four-day administration of melatonin in the late day phase advanced the N-acetyltransferase rise in 15-day old rats as compared with the rise in the vehicle-treated animals; a slight melatonin induced phase advance in 5- and 27-day-old rats was not significant. The data indicate that the newborn rat's circadian pacemaker controlling the rhythmic N-acetyltransferase rise may be entrained by exogenous melatonin. It appears, however, that the maternal melatonin transferred via milk cannot entrain the pup's pacemaker by phase advancing it, since the N-acetyltransferase rise in the pups begins earlier or at the same time as maternal melatonin production driven by the N-acetyltransferase rhythm. PMID- 9406985 TI - Melatonin cycle in the fiddler crab Uca pugilator and influence of melatonin on limb regeneration. AB - Melatonin was measured over 24 hr in the eyestalks of Uca pugilator by means of radioimmunoassay; crabs were acclimatized either to a LD 12:12 photoperiod or constant darkness. A significant peak occurred at 13.00 hr in the LD 12:12 crabs. A photophase peak in melatonin has only been reported in one other species, also a crustacean. In constant darkness, two melatonin peaks occurred, one at 16.00 hr and the other 12 hr later; these results suggest that the melatonin cycle is a true circadian rhythm. HPLC with ultraviolet-visible detection was used to confirm the identity of melatonin immunoactivity. The influence of melatonin on regeneration of the walking legs was also examined: eyestalks were either removed or left intact, and limb bud length was measured every other day for at least 17 days in control and melatonin-treated crabs (60 microg ml(-1) seawater). Melatonin significantly increased the rate of limb regeneration in both eyestalk intact and eyestalk-removed groups; this is contrary to results of regeneration studies in other phyla, in which similar melatonin concentrations inhibited regeneration. PMID- 9406986 TI - Differential inhibitory effects of melatonin analogs and three naphthalenic ligands on 2-[125I]iodomelatonin binding to chicken tissues. AB - We have compared the 50% inhibition values of 2-[125I]iodomelatonin ([125I]Mel) competition curves by melatonin and 3 naphthalenic ligands, N-[2-(7-methoxy-1 naphthyl) ethyl] cyclobutane carboxamide (S20642), N-propyl N-[2-(7-methoxy-1 naphtyl) ethyl] urea (S20753), and N-[2-(7-methoxy-1-naphthyl) ethyl] crotonamide (S20750), using membrane preparations of four tissues (lung, spleen, brain, and kidney) of the chicken simultaneously. In retired breeders, we have demonstrated that the affinities of S20642 were similar in the lung and spleen. However they were 2-fold lower in the brain and 80-fold lower in the kidney. Similar differential binding affinities to the melatonin receptors were observed in the four tissues of young male chicks. This suggests that age and sex have little influence on the differential inhibitory properties of melatonin and S20642 in the tissues studied. The addition of guanosine 5'-O-thiotriphosphate (GTPgammaS), which encouraged the uncoupling of melatonin receptor to the G protein complex, lowered the binding affinity of melatonin and S20642 in the tissues studied but their differential affinities in the four tissues were however maintained. The affinities of 5-methoxy-N-cyclopropanoyltryptamine (CPMT) in the kidney were also 5-10-fold lower than those in the lung, spleen, and brain of young male chicks. The distinctive differential affinities of melatonin, S20642, and CPMT for [125I]Mel binding sites in the chicken lung, spleen, brain, and kidney indicated that the binding sites in these tissues are heterogeneous. Our study implicated that the naphthalenic ligand S20642 may be a useful melatonin analog to distinguish melatonin receptor subtypes in tissues and a possible drug candidate worthwhile for further investigations. PMID- 9406987 TI - Influence of melatonin and serotonin on glucose-stimulated insulin release from perifused rat pancreatic islets in vitro. AB - Insulin plays a key role in the control of glucose homeostasis in mammals. Insulin secretion is regulated by a coordinated interplay of several factors. The role of the indoleamines in the control of insulin secretion has not been fully elucidated yet. The present study was addressed to investigate the function of melatonin and serotonin in the direct control of insulin secretion from the pancreatic islets. Explanted rat Langerhans' islets were treated with melatonin or serotonin while also being exposed to specific (glucose) or non-specific (KCl) stimulus either in a pulsatile or long-term manner in a perifusion system. Insulin content from the effluent tissue culture media was analyzed with RIA. Pulsatile administration of melatonin and serotonin alone did not alter the basal insulin secretion from the explanted islets even at pharmacological (5 microM) level. However, insulin response to specific (glucose) or non-specific (KCl) stimulus was significantly reduced while the islets were treated with melatonin (3 to 12 hr, 10 nM to 5 microM). This effect was reversible and repeatable. Both the start and end of the effect was rapid, evolving and disappearing within 10 min. On the other hand, under similar experimental protocol, serotonin (at 5 microM concentration) significantly enhanced both glucose and KCl stimulated insulin release. Since the effect of the non-specific stimulation (with KCl) was also altered, melatonin and serotonin seem to alter not only the release but also the synthesis of the insulin. Our data show that melatonin and serotonin have a direct effect on the insulin secretion from the pancreatic islets. PMID- 9406988 TI - Use of a newly developed technique to isolate rat pinealocytes and study the effects of adenosine agonists on melatonin production. AB - Recent studies have suggested a role for adenosine in the regulation of rat pineal melatonin synthesis. The data, however, are conflicting and therefore the aim of this study was to characterize adenosine receptors more fully in vitro by using a range of selective adenosine agonists and the adenosine antagonist 8 sulphophenyltheophylline (8-SPT). A simple method for the mechanical separation of rat pinealocytes was developed. Pinealocytes were briefly (15 min) incubated with drugs followed by a 4 hr drug-free incubation period after which melatonin concentrations in the incubation medium were measured by radioimmunoassay. The beta-adrenoceptor agonist isoprenaline gave a dose-related increase in melatonin production, demonstrating that this pinealocyte preparation technique is suitable to evaluate the effect of drugs on pineal melatonin synthesis. Our results show that adenosine, N6-(phenylisopropyl)adenosine (R-PIA) and 2-p-(2-carboxethyl) phenethylamino-5'-N-ethylcarboxamidoadenosine (CGS21680) did not affect melatonin synthesis alone or in combination with isoprenaline. However 5'-N ethylcarboxamidoadenosine (NECA) (100 microM) potentiated the stimulatory effect of isoprenaline (3 microM) on pineal melatonin production and this effect appeared to be antagonized by 8-SPT (50 microM). These results are consistent with activation by NECA of an A2b adenosine receptor. PMID- 9406989 TI - Regarding "expression of the Wilms' tumor gene (WT1) in normal hemopoiesis" by P.N. Baird and P.J. Simmons, Experimental Hematology 25:312-320 (1997) PMID- 9406990 TI - The role of polymorphonuclear neutrophils (PMNs) in clearance of granulocyte colony-stimulating factor (G-CSF) in vivo and in vitro. AB - Previous studies have suggested that in vivo granulocyte colony-stimulating factor (G-CSF) pharmacokinetics may change over time. We studied three patients treated with high-dose chemotherapy followed by autologous bone marrow transplantation (ABMT) for metastatic breast cancer after intravenous administration of recombinant human (rh) G-CSF (5 or 16 microg/kg/day). We investigated plasma G-CSF concentrations and absolute neutrophil counts (ANCs/pL) in these patients on three separate days. G-CSF plasma clearance increased with time post-ABMT with no change in the apparent volume of distribution (Vd) of G CSF. Regression analysis of G-CSF plasma clearance and ANCs revealed a linear relationship, with r2 = 0.85 (p = 0.00025). We further investigated this phenomenon in vitro by estimating pharmacokinetic parameters for rhG-CSF using a model in which polymorphonuclear neutrophils (PMNs) were incubated with rhG-CSF. We found that, at low G-CSF concentrations in vitro, there was an increase in G CSF clearance with increasing ANCs, but at higher G-CSF concentrations this relationship did not hold. We suggest that this finding resulted from aggregation and polymerization of G-CSF at high concentrations when kept at 37 degrees C for 24-48 hours in vitro. Using fluorescence staining techniques, our data suggest there are changes over time in the amount of G-CSF bound to PMNs. These changes may reflect reexpression or recycling of the G-CSF receptor, and could explain the continuing clearance of G-CSF by PMNs in vitro. The strong positive correlation between G-CSF plasma clearance and ANCs in vivo is compatible with the hypothesis that neutrophils mediate one of the major pathways for rhG-CSF clearance. PMID- 9406991 TI - Molecular and functional analysis of the Epstein-Barr virus LMP1 oncogene promoter in lymphoproliferative diseases. AB - The expression of the Epstein-Barr virus (EBV) latent membrane protein 1 (LMP1) oncogene is tightly regulated by viral and cellular factors. LMP1 is present in the majority of nasopharyngeal carcinoma tumor cells and in Reed-Sternberg cells from Hodgkin's lymphoma, in which the only EBV nuclear antigen detected is EBNA1. The aim of this study was to test whether mutations affecting LMP1 gene expression were present in lymphoproliferative disorders, and, if so, whether their presence correlated with the clinical course of the disease. For this purpose we characterized the LMP1 promoter region from seven cases including two patients with aggressive Hodgkin's disease and two with atypical lymphoproliferative syndromes. Our results show that the sequences -298 to +29 relative to the transcription start site diverged up to 9.3% when compared with the prototype EBV strain B95-8. The cAMP responsive-like element (CRE), located at positions -37 to -44, was found to be mutated in 3 of the 7 cases. Functional analysis of transfected human embryonic kidney 293 cells using the firefly luciferase reporter gene revealed that mutations within the CRE site led to a 70% mean decrease in reporter activity. Our analysis indicates that in lymphoproliferative disorders, naturally occurring LMP1 variants that exhibit weak promoter activity are still associated with clinically progressive disease. PMID- 9406992 TI - Measurement of long-term culture initiating cells (LTC-ICs) using limiting dilution: comparison of endpoints and stromal support. AB - Although much progress has been made in defining primitive cell phenotypes using flow cytometry and clonogenic assays, the direct study of marrow repopulating cells remains elusive. Long-term culture initiating cells (LTC-ICs) are arguably the most primitive human hematopoietic cells detectable by in vitro functional assays. Two endpoints have been reported for scoring LTC-ICs in limiting dilution assays. The first endpoint described was the generation of colony forming cells (CFCs) after 5 to 8 weeks of culture. An alternative method for scoring the LTC IC assay is to identify cobblestone area forming cells. In the present study, estimations of LTC-IC frequency were made by measuring both endpoints and by comparing LTC-IC frequencies measured using limiting dilution assays of normal human bone marrow stroma with the measurements for murine bone marrow stromal cell line M2-10B4. For assays established on normal human bone marrow stroma, there was an equivalence between the two endpoint measures. Likewise, there was an equivalence between the two types of stroma when scoring CFC generation after 5 weeks. However, a consistently higher frequency of LTC-ICs was estimated when scoring cobblestone areas compared with that found when scoring CFC generation on the M2-10B4 stroma (p < 0.0001). Although the murine bone marrow stromal cell line M2-10B4 remains a very useful and consistently reliable alternative to normal human bone marrow stroma, these data indicate that the LTC-IC populations defined by scoring cobblestone areas or by measuring the generation of CFCs on this cell line are, in contrast to those measured using bone marrow stroma, not identical. PMID- 9406993 TI - Unique action of an immunosuppressive agent, deoxyspergualin, on hematopoiesis in mice. AB - Deoxyspergualin (DSG) is an immunosuppresive agent of proven effectiveness in the prevention and treatment of transplant rejection; its most frequent side effect is reversible bone marrow suppression. To clarify the mechanisms of bone marrow suppression induced by DSG, we monitored the numbers of peripheral blood and marrow stem cells in C3H/HeN mice receiving 14 days of DSG injections at a highly immunosuppressive dose of 10 mg/kg/day. In the peripheral blood cells, DSG induced severe anemia and mild leukopenia because of a decrease in granulocyte counts, although these phenomena were reversible. During DSG administration, nucleated cell counts in the femur also markedly decreased, whereas the absolute numbers of various stem cells and progenitor cells, except for erythroid colony forming units (CFU-E), remained normal or increased; CD34- or c-kit-positive and lineage-negative cell levels markedly increased on the day DSG administration ceased. These findings indicate that DSG-induced anemia and leukopenia are not initiated by a generalized killing of these stem cells, but rather by a transient suppression of their ability to mature. Significantly, the severe anemia induced by DSG resembles pure red cell aplasia in humans, because there were marked decreases in peripheral reticulocytes, marrow CFU-E, and erythroblasts, with no decrease in renal erythropoietin mRNA expression. Furthermore, DSG-induced anemia was completely ameliorated by treatment with human recombinant erythropoietin. PMID- 9406994 TI - Platelet reactivity with liposome-encapsulated hemoglobin in the rat. AB - Infusion of liposome-encapsulated hemoglobin (LEH) induces a transient thrombocytopenia in rats (Rabinovici R, Rudolph AS, Ligler FS, Smith EF, III, Feuerstein G [1992] Biological responses to exchange transfusion with liposome encapsulated hemoglobin. Circ Shock 37:124). A specific mechanism such as a localization of platelets during this transient LEH-induced thrombocytopenia has not been reported previously in the literature. In this study, platelets were isolated and labeled with indium-111 (111In), then retransfused into the same animal. Fifteen minutes after the 111In platelets were retransfused and allowed to equilibrate with the blood pool, a 10% top load volume of either LEH (1.8 mL, 262 mg phospholipid/kg body weight, 92 mg hemoglobin (Hb)/kg body weight), liposome vehicle (LV) (1.8 mL, 262 mg phospholipid/kg body weight), or free bovine Hb (1.8 mL, 92 mg Hb/kg body weight) (n=6 per group) was infused. Serial blood samples were drawn to determine platelet counts by radioactivity and light scattering methods. LV and Hb demonstrated no significant changes from baseline levels in circulating platelet levels, whereas LEH caused a transient 50% decrease in 111In platelet activity 2-5 minutes postinfusion, which returned to baseline levels by 15 minutes. Platelet counts based on traditional light scattering methods were not significantly different among the three treatment groups over this same time course. Localization of these 111In platelets was monitored with a gamma scintigraphic camera. After infusion of LEH, 111In platelets rapidly moved out of the circulation and sequestered in the lungs and liver with subsequent return to the circulation by 15 minutes. In contrast, no significant changes in 111In platelet activity were noted in the lungs and liver after infusion of LV and Hb. 111In platelet activity in the spleen nearly doubled 30 minutes after Hb infusion and was significantly different than spleen 111In platelet activity in the LEH and LV groups. In vitro platelet aggregation studies were also performed to determine the direct effect of LEH, LV, and Hb on platelet aggregation. The rate of thrombin-induced aggregation did not differ among control samples and samples containing LEH, LV, or Hb; none of these agents induced platelet aggregation. We conclude that LEH-induced thrombocytopenia is associated with a transient (within minutes) sequestration of platelets in the lungs and liver, with subsequent release to the circulation within 15 minutes. Tetrameric bovine Hb is associated with increased platelet accumulation in the spleen. Light scattering methods for measuring platelet levels in blood samples containing LEH are unreliable because of particulate interference. PMID- 9406995 TI - Clonal diversity of primitive human hematopoietic progenitors following retroviral marking and long-term engraftment in immune-deficient mice. AB - The ultimate goal of human gene therapy in treating hematopoietic disorders is to insert a functional copy of the affected gene into self-renewing stem cells. The engineered pluripotent cells should then provide all lineages of corrected blood cells for the lifetime of the recipient. It is therefore important to develop methods of tracking and studying the progeny of individual human hematopoietic stem cells. Using the technique of single-colony inverse polymerase chain reaction (PCR), we assessed the clonal diversity of marked colony-forming cells that had developed from transduced human hematopoietic progenitors in a long-term xenograft system. The LN retroviral vector, which carries the neo gene, was used to individually mark human CD34+ progenitors. The marked cells were then transplanted into immune-deficient mice for periods of up to 1 year to assess their survival and retention of clonogenic capacity. Following long-term engraftment, bone marrow cells recovered from each mouse were plated in human specific colony-forming unit (CFU) assay with and without the drug G418, which selects for cells expressing the neo gene. Three weeks later, well-isolated colonies that had grown in G418 were plucked, and PCR for the neo gene was performed to confirm the presence of the vector. Inverse PCR was then performed on neo+ colonies to analyze the integration site of the LN provirus in human DNA. The clonal diversity of G418-resistant (G418R) human CFU recovered from 18 long term engrafted beige/nude/xid (bnx) mice was assessed. From one to six human hematopoietic precursors had generated all marked colony-forming progenitors (3 39) recovered from the marrow of each animal. To assess the extent of in vitro self-renewal divisions, marrow samples from 22 sets of experiments, with 2-4 mice transplanted in each set, were studied using the single-colony inverse PCR technique. Proviral integrants at identical sites were found in only two mice transplanted with cells transduced in the same flask. The presence of identical integration sites in human progenitors recovered from two mice demonstrated that a long-lived, marked cell had self-renewed in vitro before transplantation and that both daughter cells had retained the capacity to home to the bnx bone marrow and survive for 10 months. Our in vivo xenograft model and the inverse PCR technique have allowed us to identify, trace, and quantitate the clonogenic progeny of primitive human hematopoietic cells for up to 1 year after retroviral mediated transduction. PMID- 9406996 TI - Signal transduction of interleukin-6 involves tyrosine phosphorylation of multiple cytosolic proteins and activation of Src-family kinases Fyn, Hck, and Lyn in multiple myeloma cell lines. AB - Binding of interleukin-6 to its receptor (IL-6R) induces the association of the IL-6R alpha chain (IL-6Ralpha) with a 130-kDa transmembrane glycoprotein, gp130. This event activates tyrosine kinases of the Janus kinase (JAK) family and transduces signals to the cytosol or nucleus. To further characterize the biochemical mechanisms by which IL-6 promotes cell proliferation, we investigated the effects of IL-6 on the growth and transmembrane signaling of several lymphoid cell lines. In the IL-6-dependent cell line B-9, IL-6 induced a rapid, transient, and concentration-dependent tyrosine phosphorylation of several cytosolic proteins as detected by antiphosphotyrosine immunoblots. The molecular weight of major bands on sodium dodecyl sulfate-polyacrylamide gel electrophoresis was 44, 65, 70, 80, 137, 148, 184, and 190 kDa, respectively. Similar effects of IL-6 on tyrosine phosphorylation were observed in the human multiple myeloma cell line LP 1. Because JAKs were unlikely to mediate all the biological effects of IL-6, we investigated whether members of the Src family of tyrosine kinases were also activated in B-9 or LP-1 cells. IL-6 induced the activation and tyrosine phosphorylation of p59Fyn, p56/59Hck, and p56Lyn. Coprecipitation experiments with anti-Hck, anti-Lyn, anti-Fyn, and anti-gp130 antibodies revealed a physical association with gp130 of p56/59Hck and p56Lyn, but not p59Fyn, in LP-1 cells. Together, these results show for the first time that several Src kinases may become activated by IL-6 (p59Fyn, p56/59Hck, and p56Lyn) and associate with gp130 (p56/59Hck and p56Lyn). PMID- 9406997 TI - Cytokine-mediated erythroid maturation in megakaryoblastic human cell line HU-3. AB - HU-3 is a bipotential cell line derived from the bone marrow of a patient with megakaryoblastic leukemia. Continuously proliferating cells evolved from cultures supplemented with nutrient medium containing human serum and granulocyte macrophage (GM) colony-stimulating factor (CSF). Growth and viability of the HU-3 cell line was strictly dependent on the presence of GM-CSF, interleukin-3, or thrombopoietin (Tpo). Independent of the cytokine, the cells constitutively expressed a well-defined megakaryocyte phenotype, with 70-95% of the cells positive for CD4, CD34, and platelet glycoproteins Ib, IIb, and IIIa. Fewer than 10% of the cells had detectable erythroid glycophorin A. Erythropoiesis was induced in HU-3 parental cells and five clones harvested from culture medium containing GM-CSF by replacement of the growth-promoting cytokine with stem cell factor (SCF) and erythropoietin (Epo). During the first week of induction, the proliferating cells slowly acquired erythroid markers. Concomitant with a maturational growth arrest during the second week, there was a rapid accumulation of gamma and beta globin chains and benzidine reactive hemoglobin, as well as a distinct erythroid morphology. The culture declined after 12 days because of the transient effect of SCF in maintaining viability. Parental and cloned cells cultured for 7 days in Tpo-supplemented medium responded to the synergistic growth effect of SCF and Epo but were markedly suppressed in their yield of hemoglobinized cells. Recycling of the cells in GM-CSF for 4 days did not reverse the suppressive effect of Tpo. These results suggest a role for Tpo in the lineage commitment of erythromegakaryocytic progenitors by suppressing the erythroid potential. With its constitutive megakaryocyte phenotype and inducible erythroid potential, the self-renewing bipotential HU-3 cell line may represent one of the earliest stages in megakaryocytopoiesis before irreversible lineage commitment. The suppressive effect of Tpo on the erythroid potential of cloned HU 3 cells enhances the value of this cell line for deciphering the molecular and cellular events during lineage commitment of progenitor cells. PMID- 9406998 TI - Tissue-specific expression and alternative splicing of human microsomal epoxide hydrolase. AB - Human microsomal epoxide hydrolase (HYL1) plays an important role in the detoxification of environmental compounds and drugs, such as the aromatic anticonvulsants phenytoin, carbamazepine, and phenobarbital, by converting their P450-generated epoxide metabolites into nontoxic diols. Recently, we have shown that a genetic defect altering the structure and function of the HYL1 protein is unlikely to be responsible for predisposing individuals to idiosyncratic hypersensitivity reactions from anticonvulsants. To evaluate the possible involvement of regulatory mechanisms, we used 5' rapid amplification of cDNA ends (RACE) and reverse transcription polymerase chain reaction (RT-PCR) to identify and characterize HYL1 5' cDNA ends. In addition to exon 1 (E1) previously isolated from a liver cDNA library, we isolated four new exons (E1-a, E1-c, E1-d, and E1-e) from various tissues. E1 was always directly connected to exon 2 (E2) where the translation start codon is located. E1-a, E1-c E1-d, and E1-e are alternatively spliced to E2, having either E1-a or E1-a' (a truncated form of E1 a) at the 5' end of their respective transcript. Genomic data indicate that exons E1-a and E1-c are located at least 7 kb upstream from E1. Furthermore, we demonstrated a tissue-specific expression pattern for E1-containing mRNA species, whereas E1-a-containing transcripts appear to be expressed ubiquitously. Our results provide evidence that microsomal epoxide hydrolase is regulated by multiple untranslated exons flanked by tissue-specific promoters. PMID- 9406999 TI - Tissue-specific promoter usage in the D1A dopamine receptor gene in brain and kidney. AB - The D1A dopamine receptor gene consists of a short, noncoding exon 1 separated from a longer coding exon 2 by a small intron. Recently, we found that in addition to its original TATA-less promoter located upstream of exon 1, the human D1A dopamine receptor gene is transcribed in neural cells from a second strong promoter located in its intron. In the present study, we addressed the possibility that these two promoters are used for the tissue-specific regulation of the D1A gene in neuronal and renal cells. Reverse transcription polymerase chain reaction revealed that D1A transcripts in the kidneys of humans and rats lack exon 1. Transient transfection analysis of these two promoters in D1A expressing cells indicated that the upstream promoter has no detectable activity in the opossum kidney (OK) cell line, in contrast to its strong activity in two neuronal cell lines, SK-N-MC and NS20Y. On the other hand, the D1A intron promoter showed transcriptional activity both in OK cells and in neuronal cells. The activator sequence AR1, which enhances transcription from the upstream promoter in SK-N-MC and NS20Y cells, could not activate this promoter in OK cells. In addition, no protein binding to AR1 could be detected by gel mobility shift assay using nuclear extracts from either OK cells or from rat kidney tissue. These findings indicate that the differential expression of short and long D1A transcripts is due, at least in part, to the tissue-specific expression of the activator protein binding to AR1 driving transcription from the upstream promoter. Absence of this activator protein accounts for the nonfunctional D1A upstream promoter in the kidney. PMID- 9407000 TI - Use of a two-hybrid system to identify mutations in Max that confer increased affinity for Myc. AB - A yeast two-hybrid system was used to identify mutants of Max that exhibit an increased affinity for Myc. Truncated forms of the Max helix-loop-helix/leucine zipper motif (HLH/Zip) were first expressed in a two- hybrid system in which the bait protein was the HLH/Zip motif of Myc. Deletion of amino acids both amino terminal and carboxy-terminal to the leucine zipper of Max reduced Myc/Max heterodimer formation as evidenced by a 160-fold reduction in the expression of the lacZ gene. A library of partially randomized sequences encoding this minimal leucine zipper of Max was then screened using the two-hybrid system. Mutant forms of the Max leucine zipper were identified whose affinities for Myc, as measured by beta-galactosidase activity in yeast lysates, were from 8- to 200-fold greater than the wild-type Max zipper. These Max mutants were shown to interact specifically with Myc and not with wild-type Max. Of 29 mutants analyzed, all had a unique amino acid sequence. This result illustrates the value of a genetic screen in the identification of a collection of mutant forms of the Max leucine zipper whose structures would not have been predicted based on principles of structure-based design. PMID- 9407001 TI - Cloning and expression of primate Daxx cDNAs and mapping of the human gene to chromosome 6p21.3 in the MHC region. AB - Murine Daxx, a protein that binds to Fas and enhances Fas-mediated apoptosis through a signal transduction pathway involving the Jun amino-terminal kinase, was recently described. Here we report the cloning of human and monkey Daxx cDNAs, the widespread expression of Daxx mRNA in human tissues, and the mapping of the human Daxx gene to 6p21.3 in the major histocompatibility complex (MHC) region. The location of the Daxx gene, which is implicated in the pathway for deletion of autoreactive lymphocytes, in the MHC region may shed light on the genetic basis of autoimmune diseases. PMID- 9407002 TI - Effects of epidermal growth factor and estrogen on the regulation of the HMG-I/Y gene in human mammary epithelial cell lines. AB - Members of the HMG-I/Y family of high-mobility-group chromatin proteins have been demonstrated to regulate gene expression in human cells in vivo. They are thought to function as gene regulatory molecules by acting as architectural transcription factors that modulate DNA and/or chromatin structure. Numerous studies have indicated that elevated HMG-I/Y gene expression is directly correlated with more advanced cancers and with increased metastatic potential. The inducible expression of the HMG-I/Y gene was studied in two human mammary epithelial cell lines, MCF7 and Hs578T, in the presence, or absence, of either 17 beta-estradiol or epidermal growth factor (EGF). Northern blot analysis indicated that there was no increase in HMG-I/Y mRNA in the nonmetastatic MCF7 cells when they were treated with either 17 beta-estradiol or EGF. In contrast, in the highly metastatic Hs578T cell line, there is a dramatic induction of HMG-I/Y mRNA expression of up to 23-fold when the cells are treated with EGF. mRNA primer extension analysis indicated that only two (of the possible four different) transcription initiation start sites in the HMG-I/Y gene are induced by EGF treatment of the Hs578T cells. Additional experiments demonstrated that in both epithelial cell types HMG-I/Y mRNAs are very stable (tl/2 of approximately 30 hr) and that in the Hs578T cells treated with EGF the cellular concentrations of the HMG-I/Y proteins increase concurrently with the induced mRNA levels. Given that HMG-I/Y proteins are regulators of gene activity whose elevated in vivo concentrations are known to be correlated with increased metastatic potential, these data demonstrating an EGF-induced over-expression of HMG-I/Y in the highly metastatic Hs578T, but not in the nonmetastatic MCF7cells, may have important implications concerning the cellular mechanisms involved in the progression of mammary epithelial tumors. PMID- 9407003 TI - Sequence variations between two Epstein-Barr virus LMP 1 variants have no effect on the activation of NF-kappaB activity. AB - Previously, we reported that the LMP 1 gene of Epstein-Barr virus (EBV) derived from nasopharyngeal carcinoma (NPC) tissues (i.e., NLMP 1 gene) was able to transform BALB/c3T3 cells. On the other hand, LMP 1 gene of B95-8 strain (i.e., BLMP 1 gene) was not able to transform these cells (Chen et aL, 1992). Further studies indicated that a 10-amino-acid deletion in the carboxyl terminus of NLMP 1 played an important role in transformation (Li et al., 1996). In this study, we tested if this 10-amino-acid deletion affected the induction of NF-kappaB activity by LMP 1. The long terminal repeat of the human immunodeficiency virus type 1 (HIV-1 LTR) contained two copies of NF-kappaB sites and was used to construct the Luc gene-based reporter plasmid, p kappaB-Luc. Plasmid p kappaB-Luc was co-transfected with plasmids containing the NLMP 1 gene, BLMP 1 gene, and their chimeric or deletion constructs, respectively, into C-33A and BALB/c3T3 cells. The activation was then measured by the luciferase activity. Results showed that the full-length proteins induced a similar level of NF-kappaB activity, the two 3' mutants (R15delta and D4delta) still induced a relatively high level of activity, and the two 5' deletion mutants (delta3058 and delta3243) of NLMP 1 gene did not show any significant activation in C-33A cells. However, none of these LMP 1 proteins induced NF-kappaB activity in BALB/c3T3 cells. Using subcellular fractionation analysis and an immunocytostaining method, the truncated proteins of delta3058 and delta3243 were detected in the cytoplasm of the cells whereas the full-length NLMP 1 protein was located at the cytoplasmic membrane. Stable BALB/c3T3 cell clones that expressed both truncated proteins were established and then their ability to induce tumors in nude mice was examined. Data showed that both truncated NLMP 1 proteins still maintained partial transformation activity. Our results suggested that there was no direct correlation between NF-kappaB activation and transformation activity of LMP 1 in BALB/c3T3 cell transformation and that the amino-terminal membrane-spanning domain was important for maintaining both functions of LMP 1. PMID- 9407005 TI - Cyclophilin and protein disulfide isomerase genes are co-transcribed in a functionally related manner in Caenorhabditis elegans. AB - The ubiquitous enzymes peptidyl prolyl cis-trans isomerase (PPI, EC 5.2.1.8) and protein disulfide isomerase (PDI, EC 5.3.4.1) are important rate-limiting catalysts of protein-folding events in the cell. In the free-living nematode Caenorhabditis elegans, two genes encoding these enzymes (cyp-9 and pdi-1, respectively) are clustered together on chromosome III. In work described elsewhere, the encoded enzymes have been expressed as recombinant proteins and have been determined to possess in vitro PPI and PDI activity. Taken together, this organization of the two genes and the related functions of their transcripts indicate that they may be cotranscribed as a polycistronic unit, similar to bacterial operons. This study details the very close linkage of pdi-1 and cyp-9, which are in the same orientation. pdi-1 is the upstream gene, and the putative polyadenylation cleavage signal of this gene is separated from the trans-splice acceptor site of cyp-9 by only 103 bp. pdi-1 is trans-spliced by the ubiquitous nematode trans-spliced leader SL1, whereas cyp-9 was found to be predominantly trans-spliced by the "operon-specific" trans-spliced leader SL2. Similar trends in relative transcript abundance were demonstrated with synchronously produced mRNA for both genes during larval development, supporting the contention that the genes are co-expressed. Finally, reporter gene analysis provides strong evidence that both genes are controlled by a single upstream regulatory element, which directs expression of both enzymes in the hypodermal cells that synthesize the cuticle. PMID- 9407004 TI - Transforming region of 243R E1A contains two overlapping but distinct transactivation domains. AB - Conserved regions 1 and 2 as well as the amino terminus of E1A are required for the transforming activity of the E1A oncoprotein. We show here that the amino terminus of 243R E1A has transactivation activity when brought to a promoter in yeast. Recruitment to a specific promoter is essential. Mutagenesis studies correlated the transactivation function with the extreme amino terminus and the conserved region 1 of E1A. Cotransfection assays in rodent cells confirmed that two overlapping but distinguishable domains, amino acids 1-65 and 37-80, can transactivate independently when targeted to a promoter. We also observed that when recruited to the proliferating cell nuclear antigen (PCNA) promoter, the amino-terminal region was sufficient to transactivate the PCNA promoter. On the other hand, deletion of the amino terminus of E1A resulted in failure to induce PCNA expression. Fusion of VP16 with the amino-terminal-deleted E1A mutant was able to restore the ability to induce the PCNA promoter. We further show that the amino-terminal region also is required for 243R E1A to repress the transactivation mediated by a universal transactivator DBD.VP16 and DBD.E1A. This repression could be specifically relieved by overexpression of TBP but not TFIIB. In addition, we show that the amino terminus of E1A is involved in in vitro interaction with the TATA binding protein (TBP). Thus the amino-terminal transforming region of E1A may regulate cellular gene expression in species that are distant in evolution via a common mechanism, functionally targeting TBP. PMID- 9407006 TI - The Drosophila cytochrome P450 gene Cyp6a2: structure, localization, heterologous expression, and induction by phenobarbital. AB - The cytochrome P450 gene Cyp6a2 from Drosophila melanogaster is located on the right arm of chromosome 2 at position 43A1-2 and comprises two exons separated by a 69-bp intron. Phenobarbital treatment of flies leads to a rapid increase in the level of CYP6A2 mRNA and to an increased production of the CYP6A2 protein. DNA from the Cyp6a2 promoter region was functional when linked to a luciferase reporter gene and transfected into D. melanogaster Schneider cells. Moreover, a dose-dependent induction of luciferase activity by phenobarbital indicated that elements necessary for phenobarbital induction are located within 428 bp of the translation start site. Heterologous expression of the CYP6A2 protein in lepidopteran cells infected with a Cyp6a2-recombinant baculovirus was observed by Western blotting of cell lysates and by spectral characterization of the reduced CO complex of the P450. The CYP6A2 protein produced in this system metabolized aldrin and heptachlor to their epoxides and metabolized the insecticide diazinon by desulfuration to diazoxon and by oxidative ester cleavage to 2-isopropyl-4 methyl-6-hydroxypyrimidine. Metabolism in lysates of cells infected with recombinant baculovirus was greatly enhanced by the addition of purified housefly NADPH cytochrome P450 reductase and cytochrome b5. These results show that CYP6A2 catalyzes the metabolism of organophosphorus insecticides and they implicate Cyp6a2 overexpression in metabolic resistance. The Cyp6a2 gene appears to be a suitable model for a genetic analysis of the phenobarbital induction process. PMID- 9407009 TI - Molecular phylogeny of glutathione-S-transferases. AB - The glutathione-S-transferase (GST) protein superfamily is currently composed of nearly 100 sequences. This study documents a greater phylogenetic diversity of GSTs than previously realized. Parsimony and distance phylogenetic methods of GST amino acid sequences yielded virtually the same results. There appear to be at least 25 groups (families) of GST-like proteins, as different from one another as are the currently recognized classes. This diversity will require the design of a new nomenclature for this large protein superfamily. There is one well-supported large clade containing the mammalian mu, pi, and alpha classes as well as GSTs from molluscs, helminths, nematodes, and arthropods. PMID- 9407007 TI - Cloning and characterization of a novel neuropeptide Y receptor subtype in the zebrafish. AB - Neuropeptide Y (NPY), peptide YY (PYY), and pancreatic polypeptide (PP) form a family of structurally related peptides. As we have previously isolated clones for NPY and PYY from the zebrafish (Danio rerio), we wished to clone the receptors for these peptides to allow correlation of ligand and receptor distribution. We describe here the cloning and functional expression of a receptor with equally high identity to the NPY-Y1 receptor as to the recently cloned Y4/PP1 and Y6 receptors with an overall amino acid sequence identity of approximately 50%. Furthermore, the zebrafish receptor gene lacks the intron present in the coding region in vertebrate Y1 genes. These features strongly suggest that the zebrafish receptor represents a separate subtype. Hence, we have named it zYb for zebrafish Y-receptor b. (We have also discovered a unique receptor called zYa.) The zYb receptor has a binding profile that is reminiscent of Y1 with affinities for NPY and PYY in the low picomolar range, whereas affinities for Y2-selective ligands are considerably lower. It couples to adenylyl cyclase by inhibiting cAMP synthesis. Receptor mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR) in brain, eye, and intestine. The binding profile and amino acid identity show that the zebrafish zYb receptor is related to Y1 but represents a distinct subtype that is likely to be present also in mammals. PMID- 9407008 TI - Cloning and characterization of a porcine protein kinase gene and relationship to a class of heat shock proteins. AB - We have determined the genomic sequence of a porcine protein kinase (PPK) gene, including 1,844 bp upstream of the transcription initiation site. The gene spans over 19 kb and consists of 18 exons and 17 introns. The 5' regulatory region contains a characteristic heat shock element in the first intron, a weak heat shock element 1,464 bp upstream of the transcription initiation site, an atypical TATA box, and further consensus sequences typical for eukaryotic promoters such as an SP-1 binding site. Southern blot analysis indicates that PPK exists as a single-copy gene in the porcine haploid genome. The PPK gene is transcribed in all investigated tissues as shown by Northern blotting and reverse transcriptase polymerase chain reaction. Comparison of the protein and cDNA sequences of PPK to other sequences in DNA and protein databases indicates significant homology to a class of heat shock proteins, the glucose-regulated proteins (GRP94). In addition, nucleotide sequences at the 5' terminus of the PPK gene show strong homology to the GRP94 family. Domains highly conserved with human tumor rejection antigen (GP96) or glucose-regulated protein (GRP94) genes are identified within the 5' terminus and the first intron of the PPK gene. These findings suggest that these proteins are either identical or represent a family of closely related proteins. PMID- 9407010 TI - Localization of androgen receptor mRNA-containing cells in avian respiratory vocal nuclei: an in situ hybridization study. AB - Song development and song pattern formation in oscine songbirds are influenced by steroid hormones such as estrogens and androgens, and the control of vocal pattern generation is mediated via a network of interconnected vocal and respiratory nuclei. The main components of the respiratory part of the network are the expiratory and inspiratory premotor nuclei, known as retroambigualis (RAm) and the rostral ventral respiratory group (rVRG), respectively. These respiratory components play an integral role in song production either by providing the expiratory pulses of air required for each and every song syllable, or by controlling inspiration between syllables in the form of minibreaths, and between phrases for major replenishments of air. Here we analyze the distribution of androgen receptors (AR) and estrogen receptors (ER) in the midbrain and hindbrain of male and female zebra finches, and male canaries and green finches, using in situ hybridization with cRNA probes of the zebra finch AR and ER. ERmRNA was not expressed in any of the respiratory-vocal nuclei of the midbrain or hindbrain, but ARmRNA was expressed in the tracheosyringeal motor nucleus (nXIIts) and in RAm and rVRG. The size of the ARmRNA defined RAm and rVRG was similar in male and female zebra finches, but the size of ARmRNA defined nXIIts was slightly sexual dimorphic. Previously undescribed areas of ARmRNA expression outside the respiratory-vocal network in the brain stem were the nucleus semilunaris and layers 10-12 of the optic tectum. AR-mRNA expression in the respiratory-vocal nuclei of adult male songbirds, adult female zebra finches, and juvenile zebra finches suggests that the temporal pattern of learned and unlearned vocalizations is sensitive to androgen-dependent mechanisms mediated by RAm and rVRG. PMID- 9407011 TI - Identification of a phylogenetically conserved Sug1 CAD family member that is differentially expressed in the mouse nervous system. AB - We have isolated a cDNA clone from mouse, m56, that encodes a member of the Conserved ATPase-containing Domain (CAD) protein family. Sequence analysis revealed that m56 is identical to mouse mSug1/FZA-B and shares high homology with human Trip1, moth 18-56, and yeast Sug1. When examined, Sug1-like CAD proteins appear to function in the regulation of the 26S proteasome, as well as associate with members of the steroid/thyroid receptor superfamily and other transcriptional activators. m56 can complement the lethal phenotype of loss of SUG1 in yeast. We have examined the tissue distribution of m56 using Northern and Western blots, in addition to immunocytochemistry and in situ hybridization. While m56 was expressed in all tissues and cells examined, several classes of neurons, most notably in the hippocampus, olfactory bulb, and cerebellum, displayed elevated levels of m56 mRNA and protein. We also examined distribution of RNA polymerase II and 26S proteasome subunit 4 (S4) within the mouse brain by in situ hybridization. While all three genes had similar patterns of expression, there were significant differences among them. In moths, the expression of the Sug1 homolog 18-56 is dramatically up-regulated during programmed cell death. In addition, it has been previously demonstrated that the proteasome plays an essential role in the regulation of apoptosis in mammals. We examined the expression of m56 in mouse during natural and induced cell death in a variety of tissues and found no significant changes in expression. Taken together, the data presented here suggest that while m56 is a highly conserved gene that presumably plays essential but complex roles in basal and developmental processes, it may not represent a rate-limiting step in these processes. PMID- 9407012 TI - Sensorimotor pathways involved in interjoint reflex action of an insect leg. AB - Coordination of motor output between leg joints is crucial for the generation of posture and active movements in multijointed appendages of legged organisms. We investigated in the stick insect the information flow between the middle leg femoral chordotonal organ (fCO), which measures position and movement in the femur-tibia (FT) joint and the motoneuron pools supplying the next proximal leg joint, the coxa-trochanteral (CT) joint. In the inactive animal, elongation of the fCO (by flexing the FT joint) induced a depolarization in eight of nine levator trochanteris motoneurons, with a suprathreshold activation of one to three motoneurons. Motoneurons of the depressor trochanteris muscle were inhibited by fCO elongation. Relaxation signals, i.e., extension of the FT joint, activated both levator and depressor motoneurons; i.e., both antagonistic muscles were coactivated. Monosynaptic as well as polysynaptic pathways contribute to interjoint reflex actions in the stick insect leg. fCO afferents were found to induce short latency EPSPs in levator motoneurons, providing evidence for direct connections between fCO afferents and levator motoneurons. In addition, neuronal pathways via intercalated interneurons were identified that transmit sensory information from the fCO onto levator and/or depressor motoneurons. Finally, we describe two kinds of alterations in interjoint reflex action: (a) With repetitive sensory stimulation, this interjoint reflex action shows a habituation like decrease in strength. (b) In the actively moving animal, interjoint reflex action in response to fCO elongation, mimicking joint flexion, qualitatively remained the same sign, but with a marked increase in strength, indicating an increased influence of sensory signals from the FT joint onto the adjacent CT joint in the active animal. PMID- 9407013 TI - Expression and regulation of alpha1beta1 integrin in Schwann cells. AB - The interaction of cells with the extracellular matrix plays a critical role in morphogenesis and cell differentiation. To define how Schwann cells might interact with the extracellular matrix, we chose to study the expression of the laminin/collagen receptor alpha1beta1 integrin during nerve development in the rat from embryonic day 14 to maturity. We found that this integrin is expressed predominantly on mature non-myelin-forming cells and only at very low levels on myelin-forming cells. Significant levels of this integrin were not detected on Schwann cell precursors or embryonic Schwann cells in vivo. Experiments using transected and crushed sciatic nerve showed that alpha1beta1 integrin expression is regulated at least in part by axonal contact. Furthermore, Schwann cell culture experiments showed that alpha1beta1 integrin levels are strongly upregulated by transforming growth factor-beta(s) and phorbol esters. PMID- 9407014 TI - Temporal regulation of growth cone lamellar protrusion and the influence of target tissue. AB - Guided nerve fiber growth depends upon the activities of the neuronal growth cone lamellae and filopodia. Defining the dynamics of growth cone remodeling and the influences that act on it may lead to greater understanding of guided axonal growth. While there were differences in the remodeling of growth cones of nerve fibers extended from spinal cord explants and from dorsal root ganglia of Rana pipiens larvae, both types exhibited fluctuations in lamellar expanse over time to produce "lamellar cycles." We now show that these cycles are characterized by the temporal regulation of lamellar protrusion rate, the percentage of the lamellar perimeter undergoing protrusion, and invariant lamellar retraction with respect to time. Since axotomies did not abolish the lamellar cycles, the mechanism underlying cycling appears to reside at the level of the nerve fiber terminus. The previously demonstrated effects of the target tissue on growth cone remodeling appear to be due to target tissue-released factors that bind to the culture substratum, as evidenced by experiments using target tissue-conditioned medium. Further, the target tissue attenuated the fluctuations in lamellar protrusion rate during cycling, which resulted in changes in growth cone remodeling and morphology. These alterations may be related to the chemokinetic and chemotropic effects of the target on the nerve fiber extension. Thus, the process of remodeling of growth cone lamellar structures is the result of intrinsically controlled modifications in lamellar protrusion and target-based influences. PMID- 9407015 TI - Delaminating myelin membranes help seal the cut ends of severed earthworm giant axons. AB - Transected axons are often assumed to seal by collapse and fusion of the axolemmal leaflets at their cut ends. Using photomicroscopy and electronmicroscopy of fixed tissues and differential interference contrast and confocal fluorescence imaging of living tissues, we examined the proximal and distal cut ends of the pseudomyelinated medial giant axon of the earthworm, Lumbricus terrestris, at 5-60 min post-transection in physiological salines and Ca2+-free salines. In physiological salines, the axolemmal leaflets at the cut ends do not completely collapse, much less fuse, for at least 60 min post transection. In fact, the axolemma is disrupted for 20-100 microm from the cut end at 5-60 min post-transection. However, a barrier to dye diffusion is observed when hydrophilic or styryl dyes are placed in the bath at 15-30 min post transection. At 30-60 min post-transection, this barrier to dye diffusion near the cut end is formed amid an accumulation of some single-layered and many multilayered vesicles and other membranous material, much of which resembles delaminated pseudomyelin of the glial sheath. In Ca2+-free salines, this single and multilayered membranous material does not accumulate, and a dye diffusion barrier is not observed. These and other data are consistent with the hypothesis that plasmalemmal damage in eukaryotic cells is repaired by Ca2+-induced vesicles arising from invaginations or evaginations of membranes of various origin which form junctional contacts or fuse with each other and/or the plasmalemma. PMID- 9407016 TI - 17beta-estradiol attenuates CREB decline in the rat hippocampus following seizure. AB - Cyclic AMP response element-binding protein (CREB) is a transcription factor that has been implicated in the activation of a number of genes. We reported that CREB levels decline following a severe hypoglycemic episode in the hippocampus and cortex in the male rat brain. The present experiment was undertaken to investigate whether 17beta-estradiol prevents the decline in CREB-immunoreactive cells following seizure in female rats. Rats were divided into four groups: ovariectomized (OVX), ovariectomized and insulin-treated (OVX-I), estrogen replaced (E2), and estrogen-replaced and insulin-treated (E2-I). Generalized seizures were induced by injections with insulin (12.5 IU/kg, intraperitoneally) and animals were recovered by administration of glucose within 5 min of the occurrence of seizure. Control animals were injected with saline instead of insulin. All animals were perfused 90 min after recovery and the brains were processed for CREB immunoreactivity. CREB-positive neurons were counted using a computer-assisted program. Insulin treatment of OVX rats caused a significant decline in CREB-positive neurons in the CA1, CA3, and dentate gyrus compared to OVX rats. Estrogen treatment of OVX rats significantly increased CREB-positive neurons in the CA1 and dentate gyrus and attenuated the insulin-induced decline of CREB-positive neurons in all three regions compared to OVX rats. In conclusion, estrogens appear to induce CREB expression and attenuate its decline in the hippocampus following a severe hypoglycemic episode. PMID- 9407017 TI - Differences in the fate of neuronal acetylcholine receptor protein expressed in neurons and stably transfected cells. AB - Ligand-gated ion channels are structurally complex transmembrane proteins that all neurons must synthesize for rapid chemical synaptic transmission. The most abundant nicotinic acetylcholine receptor serving as a ligand-gated ion channel in the nervous system is a species that contains alpha7 subunits, binds alpha bungarotoxin, and has a high relative permeability to calcium. The ability of neurons to make such receptors was compared with that of non-neuronal cells stably transfected with an alpha7 cDNA to determine whether neuron-specific machinery is likely to aid in their assembly or stabilization. Transfected cells expressed alpha7 protein and assembled it into a species that was indistinguishable in size and pharmacology from native receptors, but much of the alpha7 protein they synthesized was rapidly degraded without becoming receptor. Neurons were not only more efficient than the best transfectants at assembling the receptors but also produced a subpopulation of receptors on the cell surface that was relatively stable and resistant to solubilization. This subpopulation, which was absent from transfected cells, may be tethered to cytoskeletal elements in the neurons. The results support the contention that neurons contain components that facilitate the production and stabilization of ligand-gated ion channels. PMID- 9407018 TI - New observations on the development of the gonadotropin-releasing hormone system in the mouse. AB - In ongoing efforts to study the ontogeny of gonadotropin-releasing hormone (GnRH) neurons, we serendipitously observed that increasing times of incubation in antibodies enhanced signal detection. Here, we describe significant differences in the early migration pattern, population dynamics, and growth cone morphology from published reports. The first immunoreactive GnRH cells were detected in the mouse at E10.75 (7.6 +/- 2.8 cells; morning after mating = E0.5), prior to the closure of the olfactory placode. Although half of these cells were in the medial wall of the olfactory pit, the other half had already initiated their migration, and approximately one quarter had reached the telencephalic vesicle. Although the migratory pattern of the GnRH cells after E11.00 was identical to that described previously, these earliest migrating cells traveled singly rather than in cords, with some reaching the presumptive preoptic area (posterior to the ganglionic eminence) by E11.75. The number of GnRH cells increased significantly (p < 0.05) to 777 +/- 183 at E11.75 and peaked at 1949.6 +/- 161.6 (p < 0.05) at E12.75. The adult population was approximately 800 cells distributed between the central nervous system (CNS) and the nasal region. Hence, the population of GnRH neurons during early development is much larger than previously appreciated; mechanisms for its decline are discussed. Neuritic extensions on the earliest GnRH neurons are short (30-50 microm) and blunt and may represent the leading edge of the moving cell. By E12.75, GnRH axons in the CNS had a ribboned or beaded morphology and increasingly more complex growth cones were noted from this time until the day of birth. The most complex growth cones were associated with apparent choice points along the axons' trajectory. By E13.75, GnRH axons were seen at the presumptive median eminence in all animals, and it was at this stage that the axons began to branch profusely. Branching, as well as the presence of growth cones, continued post-natally. These results provide further insights into the pathfinding mechanisms of GnRH cells and axons. PMID- 9407019 TI - Agrin and acetylcholine receptors are removed from abandoned synaptic sites at reinnervated frog neuromuscular junctions. AB - Changes in the distribution of agrin and acetylcholine receptors (AChRs) were examined during reinnervation and following permanent denervation as a means of understanding mechanisms controlling the distribution of these molecules. Following nerve damage in the peripheral nervous system, regenerating nerve terminals preferentially return to previous synaptic sites leading to the restoration of synaptic activity. However, not all portions of original synaptic sites are reoccupied: Some of the synaptic sites are abandoned by both the nerve terminal and the Schwann cell. Abandoned synaptic sites contain agrin, AChRs, and acetylcholinesterase (AChE) without an overlying nerve terminal or Schwann cell providing a unique location to observe changes in the distribution of these synapse-specific molecules. The distribution of anti-agrin and AChR staining at abandoned synaptic sites was altered during the process of reinnervation, changing from a dense, wide distribution to a punctate, pale pattern, and finally becoming entirely absent. Agrin and AChRs were removed from abandoned synaptic sites in reinnervated frog neuromuscular junctions, while in contralateral muscles which were permanently denervated, anti-agrin and AChR staining remained at abandoned synaptic sites. Decreasing synaptic activity during reinnervation delayed the removal of agrin and AChRs from abandoned synaptic sites. Altogether, these results support the hypothesis that synaptic activity controls a cellular mechanism that directs the removal of agrin from synaptic basal lamina and the loss of agrin leads to the dispersal of AChRs. PMID- 9407020 TI - Expression of neurotrophins and Trk receptors in the developing, adult, and regenerating avian cochlea. AB - We studied the expression of neurotrophins and their Trk receptors in the chicken cochlea. Based on in situ hybridization, brain-derived neurotrophic factor (BDNF) is the major neurotrophin there, in contrast to the mammalian cochlea, where neurotrophin-3 (NT-3) predominates. NT-3 mRNA labeling was weak and found only during a short time period in the early cochleas. During embryogenesis, BDNF mRNA was first seen in early differentiating hair cells. Afferent cochlear neurons expressed trkB mRNA from the early stages of gangliogenesis onward. In accordance, in vitro, BDNF promoted survival of dissociated neurons and stimulated neuritogenesis from ganglionic explants. High levels of BDNF mRNA in hair cells and trkB mRNA in cochlear neurons persisted in the mature cochlea. In addition, mRNA for the truncated TrkB receptor was expressed in nonneuronal cells, specifically in supporting cells, located adjacent to the site of BDNF synthesis and nerve endings. Following acoustic trauma, regenerated hair cells acquired BDNF mRNA expression at early stages of differentiation. Truncated trkB mRNA was lost from supporting cells that regenerated into hair cells. High levels of BDNF mRNA persisted in surviving hair cells and trkB mRNA in cochlear neurons after noise exposure. These results suggest that in the avian cochlea, peripheral target-derived BDNF contributes to the onset and maintenance of hearing function by supporting neuronal survival and regulating the (re)innervation process. Truncated TrkB receptors may regulate the BDNF concentration available to neurites, and they might have an important role during reinnervation. PMID- 9407021 TI - Levels of MyoD protein expression following injury of mdx and normal limb muscle are modified by thyroid hormone. AB - Thyroid hormone (T3) affects muscle development and muscle regeneration. It also interacts with the muscle regulatory gene MyoD in culture and affects myoblast proliferation. We studied the localization of MyoD protein using a well characterized polyclonal antibody for immunohistochemistry. Relative numbers of myogenic precursor cells per field were identified by their MyoD expression during muscle regeneration in normal and mdx dystrophic mice, with particular reference to the expression in mononuclear cells and myotubes at various T3 levels. In regeneration by normal muscles, relatively few MyoD+ nuclei per field were present in mononuclear cells of euthyroid and hypothyroid mice. MyoD staining of mononuclear cell nuclei was approximately doubled in fields of regenerating muscles of normal hyperthyroid compared to euthyroid mice, and was observed in precursors that appeared to be aligned before fusion into myotubes. In mdx regenerating muscle, twofold more mononuclear cells positive for MyoD were present in all three treatment groups compared to normal muscles regenerating under the same conditions. Localization was similar to the pattern in normal euthyroid mice. However, in muscles regenerating in hyperthyroid mdx mice, both mononuclear cell nuclei and centrally located nuclei in a subpopulation (about 15%) of new myotubes formed after the crush injury were intensely stained for MyoD protein. The changes observed are consistent with reports on T3-induced alteration of muscle repair, and propose a link between MyoD regulation and the accelerated differentiation during regeneration under high T3 conditions. (J Histochem Cytochem 46:59-67, 1998) PMID- 9407022 TI - Transcription units as RNA processing units. PMID- 9407023 TI - Wnt signaling: a common theme in animal development. PMID- 9407025 TI - mRNA capping enzyme is recruited to the transcription complex by phosphorylation of the RNA polymerase II carboxy-terminal domain. AB - Capping of mRNA occurs shortly after transcription initiation, preceding other mRNA processing events such as mRNA splicing and polyadenylation. To determine the mechanism of coupling between transcription and capping, we tested for a physical interaction between capping enzyme and the transcription machinery. Capping enzyme is not stably associated with basal transcription factors or the RNA polymerase II (Pol II) holoenzyme. However, capping enzyme can directly and specifically interact with the phosphorylated form of the RNA polymerase carboxy terminal domain (CTD). This association occurs in the context of the transcription initiation complex and is blocked by the CTD-kinase inhibitor H8. Furthermore, conditional truncation mutants of the Pol II CTD are lethal when combined with a capping enzyme mutant. Our results provide in vitro and in vivo evidence that capping enzyme is recruited to the transcription complex via phosphorylation of the RNA polymerase CTD. PMID- 9407024 TI - 5'-Capping enzymes are targeted to pre-mRNA by binding to the phosphorylated carboxy-terminal domain of RNA polymerase II. AB - We have investigated the role of the RNA Polymerase II (Pol II) carboxy-terminal domain (CTD) in mRNA 5' capping. Transcripts made in vivo by Pol II with a truncated CTD had a lower proportion of capped 5' ends than those made by Pol II with a full-length CTD. In addition, the enzymes responsible for cap synthesis, RNA guanylyltransferase, and RNA (guanine-7)-methyltransferase bound directly to the phosphorylated, but not to the nonphosphorylated, form of the CTD in vitro. These results suggest that: (1) Pol II-specific capping of nascent transcripts in vivo is enhanced by recruitment of the capping enzymes to the CTD and (2) capping is co-ordinated with CTD phosphorylation. PMID- 9407026 TI - Histone acetyltransferases regulate HIV-1 enhancer activity in vitro. AB - Specific inhibitors of histone deacetylase, such as trichostatin A (TSA) and trapoxin (TPX), are potent inducers of HIV-1 transcription in latently infected T cell lines. Activation of the integrated HIV-1 promoter is accompanied by the loss or rearrangement of a positioned nucleosome (nuc-1) near the viral RNA start site. Here we show that TSA strongly induces HIV-1 transcription on chromatin in vitro, concomitant with an enhancer factor-assisted increase in the level of acetylated histone H4. TSA treatment, however, did not detectably alter enhancer factor binding or the positioning of nuc-1 on the majority of the chromatin templates indicating that protein acetylation and chromatin remodeling may be limiting steps that occur only on transcriptionally competent templates, or that remodeling of nuc-1 requires additional factors. To assess the number of active chromatin templates in vitro, transcription was limited to a single round with low levels of the detergent Sarkosyl. Remarkably, HIV-1 transcription on chromatin was found to arise from a small number of active templates that can each support nearly 100 rounds of transcription, and TSA increased the number of active templates in each round. In contrast, transcription on naked DNA was limited to only a few rounds and was not responsive to TSA. We conclude that HIV 1 enhancer complexes greatly facilitate transcription reinitiation on chromatin in vitro, and act at a limiting step to promote the acetylation of histones or other transcription factors required for HIV-1 enhancer activity. PMID- 9407027 TI - Met receptor signaling is required for sensory nerve development and HGF promotes axonal growth and survival of sensory neurons. AB - The development of the nervous system is a dynamic process during which factors act in an instructive fashion to direct the differentiation and survival of neurons, and to induce axonal outgrowth, guidance to, and terminal branching within the target tissue. Here we report that mice expressing signaling mutants of the hepatocyte growth factor (HGF) receptor, the Met tyrosine kinase, show a striking reduction of sensory nerves innervating the skin of the limbs and thorax, implicating the HGF/Met system in sensory neuron development. Using in vitro assays, we find that HGF cooperates with nerve growth factor (NGF) to enhance axonal outgrowth from cultured dorsal root ganglion (DRG) neurons. HGF also enhances the neurotrophic activities of NGF in vitro, and Met receptor signaling is required for the survival of a proportion of DRG neurons in vivo. This synergism is specific for NGF but not for the related neurotrophins BDNF and NT3. By using a mild signaling mutant of Met, we have demonstrated previously that Met requires signaling via the adapter molecule Grb2 to induce proliferation of myoblasts. In contrast, the actions of HGF on sensory neurons are mediated by Met effectors distinct from Grb2. Our findings demonstrate a requirement for Met signaling in neurons during development. PMID- 9407028 TI - Nuclear hormone receptor antagonism with AP-1 by inhibition of the JNK pathway. AB - The activity of c-Jun, the major component of the transcription factor AP-1, is potentiated by amino-terminal phosphorylation on serines 63 and 73 (Ser-63/73). This phosphorylation is mediated by the Jun amino-terminal kinase (JNK) and required to recruit the transcriptional coactivator CREB-binding protein (CBP). AP-1 function is antagonized by activated members of the steroid/thyroid hormone receptor superfamily. Recently, a competition for CBP has been proposed as a mechanism for this antagonism. Here we present evidence that hormone-activated nuclear receptors prevent c-Jun phosphorylation on Ser-63/73 and, consequently, AP-1 activation, by blocking the induction of the JNK signaling cascade. Consistently, nuclear receptors also antagonize other JNK-activated transcription factors such as Elk-1 and ATF-2. Interference with the JNK signaling pathway represents a novel mechanism by which nuclear hormone receptors antagonize AP-1. This mechanism is based on the blockade of the AP-1 activation step, which is a requisite to interact with CBP. In addition to acting directly on gene transcription, regulation of the JNK cascade activity constitutes an alternative mode whereby steroids and retinoids may control cell fate and conduct their pharmacological actions as immunosupressive, anti-inflammatory, and antineoplastic agents. PMID- 9407029 TI - Mcm2 is a target of regulation by Cdc7-Dbf4 during the initiation of DNA synthesis. AB - The initiation of DNA synthesis is an important cell cycle event that defines the beginning of S phase. This critical event involves the participation of proteins whose functions are regulated by cyclin dependent protein kinases (Cdks). The Mcm2-7 proteins are a family of six conserved proteins that are essential for the initiation of DNA synthesis in all eukaryotes. In Saccharomyces cerevisiae, members of the Mcm2-7 family undergo cell cycle-specific phosphorylation. Phosphorylation of Mcm proteins at the beginning of S phase coincides with the removal of these proteins from chromatin and the onset of DNA synthesis. In this study, we identified DBF4, which encodes the regulatory subunit of a Cdk-like protein kinase Cdc7-Dbf4, in a screen for second site suppressors of mcm2-1. The dbf4 suppressor mutation restores competence to initiate DNA synthesis to the mcm2-1 mutant. Cdc7-Dbf4 interacts physically with Mcm2 and phosphorylates Mcm2 and three other members of the Mcm2-7 family in vitro. Blocking the kinase activity of Cdc7-Dbf4 at the G1-to-S phase transition also blocks the phosphorylation of Mcm2 at this defined point of the cell cycle. Taken together, our data suggest that phosphorylation of Mcm2 and probably other members of the Mcm2-7 proteins by Cdc7-Dbf4 at the G1-to-S phase transition is a critical step in the initiation of DNA synthesis at replication origins. PMID- 9407030 TI - Persistent initiation of DNA replication and chromatin-bound MCM proteins during the cell cycle in cdc6 mutants. AB - Faithful inheritance of genetic information requires that DNA be copied only once each cell cycle. Initiation of DNA replication involves the establishment of a prereplication complex (pre-RC) and subsequent activation by CDK/cyclins, converting the pre-RC to a post-RC. The origin recognition complex (ORC), Cdc6p, and the MCM proteins are required for establishing the pre-RC. We show that all six ORC subunits remain bound to chromatin throughout the cell cycle, whereas the MCM proteins cycle on and off, corresponding precisely to transitions of the RC. A newly isolated cdc6 mutant displays promiscuous initiation of DNA replication, increased nuclear DNA content, and constant MCM protein association with chromatin throughout the cell cycle. This gain-of-function cdc6 mutant ignores the negative controls imposed normally on initiation by the CDK/cyclins, suggesting that Cdc6p is a key mediator of once-per-cell-cycle control of DNA replication. PMID- 9407031 TI - Damage and replication checkpoint control in fission yeast is ensured by interactions of Crb2, a protein with BRCT motif, with Cut5 and Chk1. AB - Fission yeast Cut5/Rad4 plays a unique role in the genome maintenance as it is required for replication, replication checkpoint, and normal UV sensitivity. It is unknown, however, how Cut5 protein is linked to other checkpoint proteins, and what part it plays in replication and UV sensitivity. Here we report that Cut5 interacts with a novel checkpoint protein Crb2 and that this interaction is needed for normal genome maintenance. The carboxyl terminus of Crb2 resembles yeast Rad9 and human 53BP1 and BRCA1. Crb2 is required for checkpoint arrests induced by irradiation and polymerase mutations, but not for those induced by inhibited nucleotide supply. Upon UV damage, Crb2 is transiently modified, probably phosphorylated, with a similar timing of phosphorylation in Chk1 kinase, which is reported to restrain Cdc2 activation. Crb2 modification requires other damage-sensing checkpoint proteins but not Chk1, suggesting that Crb2 acts at the upstream of Chk1. The modified Crb2 exists as a slowly sedimenting form, whereas Crb2 in undamaged cells is in a rapidly sedimenting structure. Cut5 and Crb2 interact with Chk1 in a two-hybrid system. Moreover, moderate overexpression of Chk1 suppresses the phenotypes of cut5 and crb2 mutants. Cut5, Crb2, and Chk1 thus may form a checkpoint sensor-transmitter pathway to arrest the cell cycle. PMID- 9407032 TI - Budding yeast centromere composition and assembly as revealed by in vivo cross linking. AB - The centromere-kinetochore complex is a specialized chromatin structure that mediates bipolar attachment of replicated chromosomes to the mitotic spindle, thereby ensuring proper sister chromatid separation during anaphase. The manner in which this important multimeric structure is specified and assembled within chromatin is unknown. Using in vivo cross-linking followed by immunoprecipitation, we show that the Mif2 protein of the budding yeast Saccharomyces cerevisiae, previously implicated in centromere function by genetic criteria, resides specifically at centromeric loci in vivo. This provides definitive evidence for structural conservation between yeast and mammalian centromeres, as Mif2p shares homology with CENP-C, a mammalian centromere protein. Ndc10p and Cbf1p, previously implicated in centromere function by genetic and in vitro biochemical assays, were also found to interact with centromeric DNA in vivo. By examining Mif2p, Ndc10p, and Cbf1p association with centromeric DNA derivatives, we demonstrate the existence of centromeric subcomplexes that may correspond to assembly intermediates. Based on these observations, we provide a simple model for centromere assembly. Finally, given the sensitivity of this technique, its application to other sequence-specific protein-DNA complexes within the cell, such as origins of replication and enhancer-promoter regions, could be of significant value. PMID- 9407033 TI - A role for CKII phosphorylation of the cactus PEST domain in dorsoventral patterning of the Drosophila embryo. AB - Regulated proteolysis of Cactus, the cytoplasmic inhibitor of the Rel-related transcription factor Dorsal, is an essential step in patterning of the Drosophila embryo. Signal-induced Cactus degradation frees Dorsal for nuclear translocation on the ventral and lateral sides of the embryo, establishing zones of gene expression along the dorsoventral axis. Cactus stability is regulated by amino terminal serine residues necessary for signal responsiveness, as well as by a carboxy-terminal PEST domain. We have identified Drosophila casein kinase II (CKII) as a Cactus kinase and shown that CKII specifically phosphorylates a set of serine residues within the Cactus PEST domain. These serines are phosphorylated in vivo and are required for wild-type Cactus activity. Conversion of these serines to alanine or glutamic acid residues differentially affects the levels and activity of Cactus in embryos, but does not inhibit the binding of Cactus to Dorsal. Taken together, these data indicate that wild-type axis formation requires CKII-catalyzed phosphorylation of the Cactus PEST domain. PMID- 9407034 TI - The preference for GT-rich DNA by the yeast Rad51 protein defines a set of universal pairing sequences. AB - The Rad51 protein of Saccharomyces cerevisiae is a eukaryotic homolog of the RecA protein, the prototypic DNA strand-exchange protein of Escherichia coli. RAD51 gene function is required for efficient genetic recombination and for DNA double strand break repair. Recently, we demonstrated that RecA protein has a preferential affinity for GT-rich DNA sequences-several of which exhibit enhanced RecA protein-promoted homologous pairing activity. The fundamental similarity between the RecA and Rad51 proteins suggests that Rad51 might display an analogous bias. Using in vitro selection, here we show that the yeast Rad51 protein shares the same preference for GT-rich sequences as its prokaryotic counterpart. This bias is also manifest as an increased ability of Rad51 protein to promote the invasion of supercoiled DNA by homologous GT-rich single-stranded DNA, an activity not previously described for the eukaryotic pairing protein. We propose that the preferred utilization of GT-rich sequences is a conserved feature among all homologs of RecA protein, and that GT-rich regions are loci for increased genetic exchange in both prokaryotes and eukaryotes. PMID- 9407035 TI - Calcineurin acts through the CRZ1/TCN1-encoded transcription factor to regulate gene expression in yeast. AB - Calcineurin is a conserved Ca2+/calmodulin-dependent protein phosphatase that plays a critical role in Ca2+ signaling. We describe new components of a calcineurin-mediated response in yeast, the Ca2+-induced transcriptional activation of FKS2, which encodes a beta-1,3 glucan synthase. A 24-bp region of the FKS2 promoter was defined as sufficient to confer calcineurin-dependent transcriptional induction on a minimal promoter in response to Ca2+ and was named CDRE (for calcineurin-dependent response element). The product of CRZ1 (YNL027w) was identified as an activator of CDRE-driven transcription. Crz1p contains zinc finger motifs and binds specifically to the CDRE. Genetic analysis revealed that crz1Delta mutant cells exhibit several phenotypes similar to those of calcineurin mutants and that overexpression of CRZ1 in calcineurin mutants suppressed these phenotypes. These results suggest that Crz1p functions downstream of calcineurin to effect multiple calcineurin-dependent responses. Moreover, the calcineurin dependent transcriptional induction of FKS2 in response to Ca2+, alpha-factor, and Na+ was found to require CRZ1. In addition, we found that the calcineurin dependent transcriptional regulation of PMR2 and PMC1 required CRZ1. However, transcription of PMR2 and PMC1 was activated by only a subset of the treatments that activated FKS2 transcription. Thus, in response to multiple signals, calcineurin acts through the Crz1p transcription factor to differentially regulate the expression of several target genes in yeast. PMID- 9407036 TI - Tcn1p/Crz1p, a calcineurin-dependent transcription factor that differentially regulates gene expression in Saccharomyces cerevisiae. AB - Ca2+ signals regulate gene expression in animal and yeast cells through mechanisms involving calcineurin, a protein phosphatase activated by binding Ca2+ and calmodulin. Tcn1p, also named Crz1p, was identified as a transcription factor in yeast required for the calcineurin-dependent induction of PMC1, PMR1, PMR2A, and FKS2 which confer tolerance to high Ca2+, Mn2+, Na+, and cell wall damage, respectively. Tcn1p was not required for other calcineurin-dependent processes, such as inhibition of a vacuolar H+/Ca2+ exchanger and inhibition of a pheromone stimulated Ca2+ uptake system, suggesting that Tcn1p functions downstream of calcineurin on a branch of the calcium signaling pathway leading to gene expression. Tcn1p contains three zinc finger motifs at its carboxyl terminus resembling the DNA-binding domains of Zif268, Swi5p, and other transcription factors. When fused to the transcription activation domain of Gal4p, the carboxy terminal domain of Tcn1p directed strong calcineurin-independent expression of PMC1-lacZ and other target genes. The amino-terminal domain of Tcn1p was found to function as a calcineurin-dependent transcription activation domain when fused to the DNA-binding domain of Gal4p. This amino-terminal domain also formed Ca2+ dependent and FK506-sensitive interactions with calcineurin in the yeast two hybrid assay. These findings suggest that Tcn1p functions as a calcineurin dependent transcription factor. Interestingly, induction of Tcn1p-dependent genes was found to be differentially controlled in response to physiological Ca2+ signals generated by treatment with mating pheromone and high salt. We propose that different promoters are sensitive to variations in the strength of Ca2+ signals generated by these stimuli and to effects of other signaling pathways. PMID- 9407037 TI - The yeast HPR1 gene has a functional role in transcriptional elongation that uncovers a novel source of genome instability. AB - The yeast HPR1 gene plays an important role in genome stability, as indicated by the observation that hpr1 mutants have high frequencies of DNA repeat recombination and chromosome loss. Here we report that HPR1 is required for transcriptional elongation. Transcription driven from constitutive and regulated yeast promoters cannot elongate through the bacterial lacZ coding region in hpr1Delta cells, but progresses efficiently through other sequences such as yeast PHO5. We show that HPR1 is not required for transcription activation and that the previously reported effects of hpr1Delta on the activation of different promoters is a consequence of the incapacity of hpr1Delta cells to elongate transcription through lacZ, used as reporter. Transcriptional defects are also observed in yeast DNA sequences of hpr1Delta cells in the presence of the transcription elongation inhibitor 6-azauracil. In all cases, the blockage of transcription elongation in hpr1Delta is associated with both the high frequency of deletions and the increase in plasmid instability that we report here. Therefore, in addition to the identification of a new element involved in transcriptional elongation, our work provides evidence for a new source of genomic instability. PMID- 9407038 TI - DNA damage induces phosphorylation of the amino terminus of p53. AB - Data are presented demonstrating that DNA damage leads to specific post translational modifications of p53 protein. Using two-dimensional peptide mapping of in vivo radiolabeled p53 tryptic phosphopeptides, recombinant truncated p53 protein, and synthetic p53 tryptic peptides, a unique p53 phosphopeptide was identified after exposure of ML-1 cells to ionizing irradiation. This peptide represents the first 24 amino acids of p53 and contains three phosphorylated serine residues. A specific p53 phosphopeptide antibody identified serine-15 as one of the two serines in p53 that becomes phosphorylated following DNA damage induced by either ionizing irradiation (IR) or ultraviolet (UV) irradiation in multiple cell types. IR-induced phosphorylation of p53 does not affect the kinetics of p53 binding to or dissociating from DNA as assessed by electrophoretic mobility-shift assays. However, p53 phosphorylation induced by DNA damage correlates with enhanced transcription of downstream p53 target genes. Low levels of phosphoserine-15 p53 are detectable within 6 hr after IR in AT cells, whereas lymphoblasts from normal individuals exhibit this modification within 1 hr. In contrast, phosphorylation of p53 on serine-15 is similar in normal and AT cells after UV irradiation. Our results indicate that p53 is phosphorylated in response to DNA damage, that this de novo phosphorylation may be involved in the subsequent induction and activation of p53, and that although ATM affects the kinetics of p53 phosphorylation after IR, it is not absolutely required for phosphorylation of p53 on serine-15. PMID- 9407040 TI - Cotranslational protein folding. PMID- 9407039 TI - Requirement for NF-kappaB in osteoclast and B-cell development. AB - NF-kappaB is a family of related, dimeric transcription factors that are readily activated in cells by signals associated with stress or pathogens. These factors are critical to host defense, as demonstrated previously with mice deficient in individual subunits of NF-kappaB. We have generated mice deficient in both the p50 and p52 subunits of NF-kappaB to reveal critical functions that may be shared by these two highly homologous proteins. We now demonstrate that unlike the respective single knockout mice, the p50/p52 double knockout mice fail to generate mature osteoclasts and B cells, apparently because of defects that track with these lineages in adoptive transfer experiments. Furthermore, these mice present markedly impaired thymic and splenic architectures and impaired macrophage functions. The blocks in osteoclast and B-cell maturation were unexpected. Lack of mature osteoclasts caused severe osteopetrosis, a family of diseases characterized by impaired osteoclastic bone resorption. These findings now establish critical roles for NF-kappaB in development and expand its repertoire of roles in the physiology of differentiated hematopoietic cells. PMID- 9407041 TI - Regulation of cell migration by the calcium-dependent protease calpain. AB - Integrin receptors play an important role during cell migration by mediating linkages and transmitting forces between the extracellular matrix and the actin cytoskeleton. The mechanisms by which these linkages are regulated and released during migration are not well understood. We show here that cell-permeable inhibitors of the calcium-dependent protease calpain inhibit both beta1 and beta3 integrin-mediated cell migration. Calpain inhibition specifically stabilizes peripheral focal adhesions, increases adhesiveness, and decreases the rate of cell detachment. Furthermore, these inhibitors alter the fate of integrin receptors at the rear of the cell during migration. A Chinese hamster ovary cell line expressing low levels of calpain I also shows reduced migration rates with similar morphological changes, further implicating calpain in this process. Taken together, the data suggest that calpain inhibition modulates cell migration by stabilizing cytoskeletal linkages and decreasing the rate of retraction of the cell's rear. Inhibiting calpain-mediated proteolysis may therefore be a potential therapeutic approach to control pathological cell migration such as tumor metastasis. PMID- 9407042 TI - A novel gene, hKCa4, encodes the calcium-activated potassium channel in human T lymphocytes. AB - We have isolated a novel gene, hKCa4, encoding an intermediate conductance, calcium-activated potassium channel from a human lymph node library. The translated protein comprises 427 amino acids, has six transmembrane segments, S1 S6, and a pore motif between S5 and S6. hKCa4 shares 41-42% similarity at the amino acid level with three small conductance calcium-activated potassium channels cloned from brain. Northern blot analysis of primary human T lymphocytes reveals a 2.2-kilobase transcript that is highly up-regulated in activated compared with resting cells, concomitant with an increase in KCa current. hKCa4 transcript is also detected by Northern blots or by polymerase chain reaction in placenta, prostate, thymus, spleen, colon, and many cell lines of hematopoietic origin. Patch-clamp recordings of hKCa4-transfected HEK 293 cells reveal a large voltage-independent, inwardly rectifying potassium current that is blocked by externally applied tetraethylammonium (Kd = 30 +/- 7 mM), charybdotoxin (Kd = 10 +/- 1 nM), and clotrimazole (Kd = 387 +/- 34 nM), but is resistant to apamin, iberiotoxin, kaliotoxin, scyllatoxin (Kd > 1 microM), and margatoxin (Kd > 100 nM). Single hKCa4 channels have a conductance of 33 +/- 2 picosiemens in symmetrical potassium solutions. The channel is activated by intracellular calcium (Kd = 270 +/- 8 nM) with a highly cooperative interaction of approximately three calcium ions per channel. These properties of the cloned channel are very similar to those reported for the native KCa channel in activated human T lymphocytes, indicating that hKCa4 encodes this channel type. PMID- 9407043 TI - Identification of the Ca2+/calmodulin-dependent protein kinase II regulatory phosphorylation site in the alpha-amino-3-hydroxyl-5-methyl-4-isoxazole propionate-type glutamate receptor. AB - Ca2+/CaM-dependent protein kinase II (CaM-KII) can phosphorylate and potentiate responses of alpha-amino3-hydroxyl-5-methyl-4-isoxazole-propionate-type glutamate receptors in a number of systems, and recent studies implicate this mechanism in long term potentiation, a cellular model of learning and memory. In this study we have identified this CaM-KII regulatory site using deletion and site-specific mutants of glutamate receptor 1 (GluR1). Only mutations affecting Ser831 altered the 32P peptide maps of GluR1 from HEK-293 cells co-expressing an activated CaM KII. Likewise, when CaM-KII was infused into cells expressing GluR1, the Ser831 to Ala mutant failed to show potentiation of the GluR1 current. The Ser831 site is specific to GluR1, and CaM-KII did not phosphorylate or potentiate current in cells expressing GluR2, emphasizing the importance of the GluR1 subunit in this regulatory mechanism. Because Ser831 has previously been identified as a protein kinase C phosphorylation site (Roche, K. W., O'Brien, R. J., Mammen, A. L., Bernhardt, J., and Huganir, R. L. (1996) Neuron 16, 1179-1188), this raises the possibility of synergistic interactions between CaM-KII and protein kinase C in regulating synaptic plasticity. PMID- 9407044 TI - Phosphorylation of human CDC25B phosphatase by CDK1-cyclin A triggers its proteasome-dependent degradation. AB - In eukaryotes the activity of CDK1 (CDC2), a cyclin-dependent kinase that initiates the structural changes that culminate in the segregation of chromosomes at mitosis, is regulated by the synergistic and opposing activities of a cascade of kinases and phosphatases. Dephosphorylation of threonine 14 and tyrosine 15 of CDK1 by the CDC25 phosphatases is a key step in the activation of the CDK1-cyclin B protein kinase. Little is currently known about the role and the regulation of CDC25B. Here we report in vitro and in vivo data that indicate that CDC25B is degraded by the proteasome. This degradation is dependent upon phosphorylation by the CDK1-cyclin A complex but not by CDK1-cyclin B. These results indicate that CDK1-cyclin A phosphorylation targets CDC25B for degradation and that this might be an important component of cell cycle regulation at the G2/M transition. PMID- 9407045 TI - On the formation and reactivity of compound I of the His-64 myoglobin mutants. AB - Myoglobin (Mb) catalyzes various two-electron oxidations; however, ferryl porphyrin cation radical equivalent to peroxidase compound I has not been identified yet. Distal histidine mutants of sperm whale Mb (His-64 --> Ala, Ser, and Leu) afford an apparent intermediate followed by the formation of a ferryl heme (Mb-II) in the reaction with m-chloroperbenzoic acid. Because the intermediate exhibits characteristic absorption spectrum of compound I and bears two electron oxidizing equivalents above the ferric state, we have assigned the species as compound I of myoglobin (Mb-I). Although we have recently observed compound I of the F43H/H64L Mb mutant, F43H and wild type Mb react with m chloroperbenzoic acid to give Mb-II without any accumulation of Mb-I. The results unambiguously indicate that His-64 plays a key role in destabilizing wild type Mb I. Furthermore, Mb-I is found to be capable of performing two-electron oxidation of styrene, thioanisole, and H2O2. PMID- 9407046 TI - Inhibition of nuclear factor-kappaB activity is involved in E1A-mediated sensitization of radiation-induced apoptosis. AB - The adenoviral E1A protein has been implicated in the potentiation of apoptosis induced by various external stimuli, but the exact mechanism of that potentiation is not clear. In this study, we compared the sensitivity to ionizing gamma irradiation of E1A transfectants with that of parental cells in a human ovarian cancer cell line (SKOV3.ip1); we found that the E1A transfectants became sensitive to radiation-induced apoptosis. Recently, activation of the transcription factor nuclear factor-kappaB (NF-kappaB) has been shown to play a key role in the anti-apoptotic pathway of radiation-induced apoptosis. In an attempt to determine whether NF-kappaB was involved in the E1A-mediated sensitization of radiation-induced apoptosis, we found that radiation-induced activation of NF-kappaB occurred in the parental cells but was blocked in the E1A transfectants. Furthermore, parental cells cotransfected with NF-kappaB and E1A were better protected from undergoing apoptosis upon irradiation than those transfected with E1A alone. Thus, our results suggest that inhibition of NF kappaB activation by E1A is a plausible mechanism for E1A-mediated sensitization of radiation-induced apoptosis. PMID- 9407047 TI - Genomic cloning of hGSTP1*C, an allelic human Pi class glutathione S-transferase gene variant and functional characterization of its retinoic acid response elements. AB - The complete hGSTP1*C, consisting of 7 exons and 6 introns contained in 3116 base pairs, was isolated from a cosmid genomic library of a glioblastoma multiforme cell line. Although the promoter of hGSTP1*C was identical to that of the previously reported GST-Pi gene, several of its structural features had not been previously described. These include several nucleotide transitions and transversions. Transitions of A --> G at +1404 and C --> T at +2294 in exons 5 and 6, respectively, changed codons Ile104 to Val104 and Ala113 to Val113. The gene also contained a guanine insertion at +51 in the insulin response element in intron 1 and six tandem repeats and one palindromic retinoic acid response element (RARE) consensus half-sites, A(G)GG(T)TC(G)A in intron 5. Retinoic acid (RA) treatment increased GST-Pi gene expression significantly in MGR3 cells. GST Pi gene constructs with and without RARE deletion were used to show the RARE requirement for GST-Pi gene induction by RA. The isolation of the hGSTP1*C gene and the evidence that it contains functional RAREs should contribute to a better understanding of the molecular regulation of the GST-Pi gene in human cells. PMID- 9407048 TI - Analogous F-actin binding by cofilin and gelsolin segment 2 substantiates their structural relationship. AB - Cofilin is representative for a family of low molecular weight actin filament binding and depolymerizing proteins. Recently the three-dimensional structure of yeast cofilin and of the cofilin homologs destrin and actophorin were resolved, and a striking similarity to segments of gelsolin and related proteins was observed (Hatanaka, H., Ogura, K., Moriyama, K., Ichikawa, S., Yahara, I., and Inagaka, F. (1996) Cell 85, 1047-1055; Fedorov, A. A., Lappalainen, P., Fedorov, E. V., Drubin, D. G., and Almo, S. C. (1997) Nat. Struct. Biol. 4, 366-369; Leonard, S. A., Gittis, A. G., Petrella, E. C., Pollard, T. D., and Lattman, E. E. (1997) Nat. Struct. Biol. 4, 369-373). Using peptide mimetics, we show that the actin binding site stretches over the entire cofilin alpha-helix 112-128. In addition, we demonstrate that cofilin and its actin binding peptide compete with gelsolin segments 2-3 for binding to actin filaments. Based on these competition data, we propose that cofilin and segment 2 of gelsolin use a common structural topology to bind to actin and probably share a similar target site on the filament. This adds a functional dimension to their reported structural homology, and this F-actin binding mode provides a basis to further enlighten the effect of members of the cofilin family on actin filament dynamics. PMID- 9407049 TI - Nuclear factor I family members regulate the transcription of surfactant protein C. AB - Transcription of the surfactant protein-C (SP-C) gene is restricted to Type II epithelial cells in the adult lung. We have shown previously that the 0.32 kilobase pair (kb) mouse SP-C promoter is functional in transient transfection assays of the lung epithelial cell-derived cell line, MLE-15, and that thyroid transcription factor 1 (TTF-1) transactivates promoter activity. The 0.32-kb SP-C promoter can be separated into a proximal promoter region (-230 to +18) and an enhancer region (-318 to -230). Three DNase I footprints were mapped in the promoter region (C1 through C3) and two in the enhancer region (C4 and C5). We now show that nuclear factor I (NFI) family members bind to both individual NFI half-sites in footprints C1, C3, and C5, and to a composite site in footprint C4 by competition gel retardation and antibody supershift analyses. Mutational analysis of the 0.32-kb mouse SP-C promoter and transient transfection of MLE-15 cells demonstrated that the NFI binding sites are required for promoter activity in this cell type. Site-specific mutation of the proximal or distal NFI sites drastically reduced transactivation by a co-transfected NFI-A expression vector in HeLa cells. These data indicate that NFI family member(s), binding to sites in both the promoter and enhancer regions, regulate SP-C gene expression in a process independent of TTF-1. PMID- 9407050 TI - Relative chlorinating, nitrating, and oxidizing capabilities of neutrophils determined with phagocytosable probes. AB - The capabilities of stimulated neutrophils to initiate intraphagosomal and extracellular chlorination, nitration, and other oxidative reactions has been evaluated using a fluorescent particle and soluble phenolic compounds as target molecules. Neutrophils activated by the soluble stimulus, phorbol myristate acetate, both chlorinated fluorescein that was covalently attached to polyacrylamide microspheres and initiated tyrosine dimerization. When nitrite ion was present at millimolar concentration levels in the medium, nitration of the phenolic rings also occurred; the relative extent of nitration increased as the nitrite concentration was increased. Myeloperoxidase (MPO) also catalyzed nitration and chlorination of fluorescein and the fluorescein-conjugated particles in cell-free solutions; the relative nitration yields increased with increasing [NO2-]/[Cl-] ratios. Nitration did not involve intermediary formation of nitrating agents derived from reaction between MPO-generated HOCl and NO2- because this reaction also occurred in chloride-free media and direct addition of HOCl to solutions containing NO2- and fluorescein gave only chlorinated products. In marked contrast to these extracellular reactions, intraphagosomal nitration of the fluorescein-conjugated particles could not be detected (even at [NO2-] as high as 0.1 M), whereas chlorination of the probe was extensive. These data indicate that intraphagosomal aromatic nitration in neutrophils is negligible, although extracellular nitration of phenolic compounds by secreted MPO could occur at physiological concentration levels of NO2-. PMID- 9407051 TI - Nitric oxide inactivates NADPH oxidase in pig neutrophils by inhibiting its assembling process. AB - The effects of nitric oxide (NO) on superoxide (O-2) generation of the NADPH oxidase in pig neutrophils were studied. NO dose-dependently suppressed O-2 generation of both neutrophil NADPH oxidase and reconstituted NADPH oxidase. Effects of NO on NADPH-binding site and the redox centers including FAD and low spin heme in cytochrome b558 and the electron transfer rates from NADPH to heme via FAD were examined under anaerobic conditions. Both reaction rates and the Km value for NADPH were unchanged by NO. Visible and EPR spectra of cytochrome b558 showed that the structure of heme was unchanged by NO, indicating that NO does not affect the redox centers of the oxidase. In reconstituted NADPH oxidase system, NO did not inhibit O-2 generation of the oxidase when added after activation. The addition of NO to the membrane component or the cytosol component inhibited the activity by 24.0 +/- 5.3 or 37.4 +/- 7.1%, respectively. The addition of NO during the activation process or to the cytosol component simultaneously with myristate inhibited the activity by 74.0 +/- 5.2 or 70.0 +/- 8.3%, respectively, suggesting that cytosol protein(s) treated with myristate becomes susceptible to NO. Peroxynitrite did not interfere with O-2 generation. PMID- 9407052 TI - Heat shock increases the association of binding protein-1 with initiation factor 4E. AB - The effects of heat shock on the regulation of the cap-binding initiation factor 4E (eIF4E) and its inhibitory binding protein, 4E-BP1, have been examined in Chinese hamster ovary cells and in cardiac myocytes. Heat shock increased the association between eIF4E and 4E-BP1, and this was associated with a dephosphorylation of 4E-BP1. These effects did not appear to be due wholly to decreased activity of the p70 S6 kinase pathway, which is implicated in the control of 4E-BP1, and they were not mediated by the stress-activated p38 microtubule-associated protein kinase pathway. Increased binding of 4E-BP1 to eIF4E correlated with a decrease in the amount of eIF4G which co-purified with the latter. This could account for the previously observed impairment of eIF4F function during heat shock, and, since heat shock protein mRNAs are believed to be relatively cap-independent, could provide a mechanism for the selective up regulation of the synthesis of heat shock proteins and other stress proteins during heat shock. PMID- 9407053 TI - Hrs is associated with STAM, a signal-transducing adaptor molecule. Its suppressive effect on cytokine-induced cell growth. AB - We previously reported a new type of signal-transducing adaptor molecule, STAM, which was shown to be involved in cytokine-mediated intracellular signal transduction. In this study, we molecularly cloned a 110-kDa phosphotyrosine protein inducible by stimulation with interleukin 2 (IL-2). The 110-kDa molecule was found to be a human counterpart of mouse Hrs (hepatocyte growth factor regulated tyrosine kinase substrate) and to be associated with STAM. Tyrosine phosphorylation of Hrs is induced rapidly after stimulation with IL-2 and granulocyte-macrophage colony-stimulating factor as well as hepatocyte growth factor. The mutual association sites of Hrs and STAM include highly conserved coiled-coil sequences, suggesting that their association is mediated by the coiled-coil structures. Exogenous introduction of the wild-type Hrs significantly suppressed DNA synthesis upon stimulation with IL-2 and granulocyte-macrophage colony-stimulating factor, while the Hrs mutant deleted of the STAM-binding site lost such suppressive ability. These results suggest that Hrs counteracts the STAM function which is critical for cell growth signaling mediated by the cytokines. PMID- 9407054 TI - Resistance to endotoxic shock in phospholipase A2 receptor-deficient mice. AB - Mammals possess various types of secretory phospholipase A2, which differ in the primary structure and tissue distribution. The phosholipase A2 receptor (PLA2R) recognizes group IB phospholipase A2 (PLA2-IB) and mediates the PLA2-IB-induced biological responses in non-digestive organs, including eicosanoid production and contraction of airway smooth muscles. In this study, we generated PLA2R-deficient mice to define its biological roles further. These mice are viable, fertile, and without evident histopathological abnormalities. There was no difference in the clearance of circulating PLA2-IB between wild-type and mutant mice. After challenge with bacterial lipopolysaccharide (LPS), PLA2R-deficient mice exhibited longer survival than wild-type mice. The mutant mice were also resistant to lethal effects of exogenous PLA2-IB after sensitization with sublethal dose of LPS. The plasma levels of tumor necrosis factor-alpha and interleukin-1beta elevated after LPS treatment were significantly reduced in mutant mice compared with wild-type mice. These findings suggest a potential role of PLA2R in the progression of endotoxic shock. PMID- 9407055 TI - Aortic smooth muscle cells interact with tenascin-C through its fibrinogen-like domain. AB - The extracellular matrix protein tenascin-C is a multidomain protein that regulates cell adhesion. We used two different smooth muscle cell subtypes derived from adult and newborn rat aorta to investigate the interaction of tenascin-C or its various domains with these cells using an adhesion assay. Newborn cells were three times more adherent to tenascin-C than adult cells. Tenascin C-adhering cells remained round, whereas they spread rapidly on a fibronectin substrate. Adhesion assays showed the interaction between tenascin-C and newborn cells to be predominantly RGD-independent. Mg2+ increased newborn cell adhesion to tenascin-C in a concentration-dependent manner, whereas Ca2+ had no effect. To analyze the structure-function relationships of different domains of tenascin-C, we used recombinant full-length fibronectin-like and fibrinogen like domains and various subdomains corresponding to the alternatively spliced regions of tenascin-C. The cells adhered to the fibrinogen-like domain but not to the fibronectin-like domain or its subdomains. As with the intact tenascin-C molecule, adherent cells remained round, and the Mg2+, but not Ca2+, promoted this interaction. The interaction of cells with the fibrinogen-like region was further mapped to a 30-amino acid peptide located near the carboxyl-terminal part of the tenascin-C molecule. The same 30-amino acid peptide was active in promoting cell migration. Our results provide a basis for understanding the mechanism of interaction of tenascin-C with smooth muscle cells and a framework for isolating membrane binding sites that mediate the cellular responses to this molecule. PMID- 9407056 TI - Carbon monoxide controls the proliferation of hypoxic vascular smooth muscle cells. AB - Excess vascular smooth muscle cell (VSMC) proliferation and contractility are key events in the pathophysiology of vascular disorders induced by hypoxia. We have recently reported that carbon monoxide (CO), produced by VSMC under conditions of hypoxia, can be a modulator of cGMP levels in both endothelial and smooth muscle cells. In this respect, some of the physiologic effects of CO in the vasculature parallel those of nitric oxide (NO), a well characterized regulator of vascular tone. We report here that under hypoxia, VSMC-derived CO is an important regulator of VSMC proliferation. Inhibiting CO formation or scavenging CO with hemoglobin increased VSMC proliferation in response to serum or to mitogens such as endothelin, whereas increasing CO production or exposing cells to exogenous CO lead to a markedly attenuated growth response. The effects of CO on VSMC proliferation correlated with changes in E2F-1 expression, the prototype member of a family of transcription factors that participate in the control of cell cycle progression. CO significantly suppressed E2F-1 expression, whereas, removal of CO from the cultures with hemoglobin lead to increased E2F-1 gene transcription, mRNA, and protein production as well as mRNA levels of c-myc, a target gene of E2F-1. Moreover, the actions of CO were mediated by the second messenger molecule, cGMP. Limiting VSMC growth by increasing the release of CO may represent a key event in the body's compensatory responses to hypoxia. PMID- 9407057 TI - Role of the prohormone convertase PC3 in the processing of proglucagon to glucagon-like peptide 1. AB - Proglucagon is processed differentially in pancreatic alpha-cells and intestinal endocrine L cells to release either glucagon or glucagon-like peptide-1-(7 36amide) (tGLP-1), two peptide hormones with opposing biological actions. Previous studies have demonstrated that the prohormone convertase PC2 is responsible for the processing of proglucagon to glucagon, and have suggested that the related endoprotease PC3 is involved in the formation of tGLP-1. To understand better the biosynthetic pathway of tGLP-1, proglucagon processing was studied in the mouse pituitary cell line AtT-20, a cell line that mimics the intestinal pathway of proglucagon processing and in the rat insulinoma cell line INS-1. In both of these cell lines, proglucagon was initially cleaved to glicentin and the major proglucagon fragment (MPGF) at the interdomain site Lys70 Arg71. In both cell lines, MPGF was cleaved successively at the monobasic site Arg77 and then at the dibasic site Arg109-Arg110, thus releasing tGLP-1, the cleavages being less extensive in INS-1 cells. Glicentin was completely processed to glucagon in INS-1 cells, but was partially converted to oxyntomodulin and very low levels of glucagon in AtT-20 cells in the face of generation of tGLP-1. Adenovirus-mediated co-expression of PC3 and proglucagon in GH4C1 cells (normally expressing no PC2 or PC3) resulted in the formation of tGLP-1, glicentin, and oxyntomodulin, but no glucagon. When expressed in alphaTC1-6 (transformed pancreatic alpha-cells) or in rat primary pancreatic alpha-cells in culture, PC3 converted MPGF to tGLP-1. Finally, GLP-1-(1-37) was cleaved to tGLP-1 in vitro by purified recombinant PC3. Taken together, these results indicate that PC3 has the same specificity as the convertase that is responsible for the processing of proglucagon to tGLP-1, glicentin and oxyntomodulin in the intestinal L cell, and it is concluded that this enzyme is thus able to act alone in this processing pathway. PMID- 9407058 TI - Multimer formation and ligand recognition by the long pentraxin PTX3. Similarities and differences with the short pentraxins C-reactive protein and serum amyloid P component. AB - PTX3 is a prototypic long pentraxin consisting of a C-terminal 203-amino acid pentraxin-like domain coupled with an N-terminal 178-amino acid unrelated portion. The present study was designed to characterize the structure and ligand binding properties of human PTX3, in comparison with the classical pentraxins C reactive protein and serum amyloid P component. Sequencing of Chinese hamster ovary cell-expressed PTX3 revealed that the mature secreted protein starts at residue 18 (Glu). Lectin binding and treatment with N-glycosidase F showed that PTX3 is N-glycosylated, sugars accounting for 5 kDa of the monomer mass (45 kDa). Circular dichroism analysis indicated that the protein consists predominantly of beta-sheets with a minor alpha-helical component. While in gel filtration the protein is eluted with a molecular mass of congruent with900 kDa, gel electrophoresis using nondenaturing, nonreducing conditions revealed that PTX3 forms multimers predominantly of 440 kDa apparent molecular mass, corresponding to decamers, and that disulfide bonds are required for multimer formation. The ligand binding properties of PTX3 were then examined. As predicted based on modeling, inductive coupled plasma/atomic emission spectroscopy showed that PTX3 does not have coordinated Ca2+. Unlike the classical pentraxins CRP and SAP, PTX3 did not bind phosphoethanolamine, phosphocholine, or high pyruvate agarose. PTX3 in solution, bound to immobilized C1q, but not C1s, and, reciprocally, C1q bound to immobilized PTX3. Binding of PTX3 to C1q is specific and saturable with a Kd 7.4 x 10(-8) M as determined by solid phase binding assay. The Chinese hamster ovary cell-expressed pentraxin domain bound C1q when multimerized. Thus, as predicted on the basis of computer modeling, the prototypic long pentraxin PTX3 forms multimers, which differ from those formed by classical pentraxins in terms of protomer composition and requirement for disulfide bonds, and does not recognize CRP/SAP ligands. The capacity to bind C1q, mediated by the pentraxin domain, is consistent with the view that PTX3, produced in tissues by endothelial cells or macrophages in response to interleukin-1 and tumor necrosis factor, may act as a local regulator of innate immunity. PMID- 9407059 TI - Expression of the human aryl hydrocarbon receptor complex in yeast. Activation of transcription by indole compounds. AB - The human aryl hydrocarbon receptor (AHR) and aryl hydrocarbon receptor nuclear translocator protein (ARNT) were coexpressed in the yeast Saccharomyces cerevisiae to create a system for the study of this heterodimeric transcription factor. Specific transcriptional activation mediated by AHR/ARNT heterodimer, which is a functional indicator of receptor expression, was assessed by beta galactosidase activity produced from a reporter plasmid. Yeast expressing AHR and ARNT displayed constitutive transcriptional activity that was not augmented by addition of AHR agonists in strains that required exogenous tryptophan for viability. In contrast, strains with an intact pathway for tryptophan biosynthesis responded to AHR agonists and had lower levels of background beta galactosidase activity. Hexachlorobenzene, benzo(a)pyrene, and beta naphthoflavone were effective AHR agonists in the yeast system, and had EC50 values of 200, 40, and 20 nM, respectively, for beta-galactosidase activity induction. Tryptophan, indole, indole acetic acid, and tryptamine activated transcription in yeast coexpressing AHR and ARNT (EC50 values approximately 300 microM). Indole-3-carbinol was an exceptionally potent AHR agonist (EC50 approximately 10 microM) in yeast. This yeast system is useful for the study of AHR/ARNT protein complexes, and may be generally applicable to the investigation of other multiprotein complexes. PMID- 9407060 TI - Structural determinants required for the interaction between Rho GTPase and the GTPase-activating domain of p190. AB - The Rho family small GTP-binding proteins are subjected to regulation by Rho GTPase-activating proteins (GAPs) in the course of transmitting diverse intracellular signals. To understand the mechanism of GAP-catalyzed GTP hydrolysis of Rho GTPases, we have studied the interaction between RhoA and p190, the RasGAP binding phosphoprotein which has been implicated as a Rho-specific GAP, by delineating the structural determinants of RhoA and p190 GAP domain (p190GD) that are involved in their functional coupling. Besides the conserved residues Tyr34, Thr37, and Phe39 in the switch I region of RhoA which are required for p190GD interaction, chimeras made between RhoA and Cdc42, a close relative of RhoA with which p190GD interacts 50-fold less efficiently, revealed that residues outside the switch I and neighboring regions of RhoA, residues 85 122 in particular, contain the major p190GD-specifying determinant(s). Mutation of the unique Asp90 of RhoA in this region mostly abolished p190GD stimulation, whereas the corresponding reverse mutation of Cdc42 (S88D) was able to respond to p190GD-catalysis similarly as RhoA. Further kinetic analysis of these mutants provided evidence that Asp90 of RhoA contributes primarily to the specific binding interaction with p190GD. On the other hand, two charged residues of p190GD, Arg1283 and Lys1321, which are located in the putative G-protein binding helix pocket of GAP domain, were found to be involved in different aspects of interaction with RhoA. The R1283L mutant of p190GD lost GAP activity but retained the ability to bind to RhoA, while K1321A failed to stimulate and to bind to RhoA. These results indicate that residue Asp90 constitutes the second GAP interactive site in RhoA which is mostly responsible for conferring p190GD specificity, and suggest that the role of p190GD in the GTPase reaction of RhoA is in part to supply active site residue Arg1283 for efficient catalysis. PMID- 9407061 TI - A far upstream cis-element is required for Wilms' tumor-1 (WT1) gene expression in renal cell culture. AB - To identify novel cis-regulatory elements responsible for the tissue-restricted expression pattern of the Wilms' tumor-1 (WT1) gene, we mapped a total of 11 DNase I-hypersensitive sites in the 5'-flanking region and first intron of the human gene, six of which were specific for WT1 expressing cell lines. A 1.4 kilobase (kb) fragment from the mouse wt1 5'-flanking region contained cross hybridizing sequence with significant homology to a region of DNase I hypersensitivity in the human WT1 gene which bound to nuclear matrix in human fetal kidney 293 cells. None of the DNase I-hypersensitive sites/matrix attachment regions, either alone or in combination, were sufficient for tissue specific WT1 expression in transient and stably transfected cell lines. However, stable transfection of an approximately 620-kb yeast artificial chromosome (YAC) that carried the entire mouse wt1 locus into 293 cells resulted in wt1 (trans)gene expression at a level of approximately 30% of the endogenous human gene. Deletion of the 1.4-kb cross-hybridizing mouse fragment, located approximately 15 kb upstream of the transcription start site, caused complete loss of wt1 gene expression in the YAC-transfected 293 cells. In summary, we have identified a far upstream element that contains a region of DNase I hypersensitivity and that binds to nuclear matrix. This element includes phylogenetically conserved sequence and is required, although not sufficient, for mouse wt1 gene expression in human fetal kidney cells in culture. PMID- 9407062 TI - alpha1-Adrenergic stimulation potentiates the thermogenic action of beta3 adrenoreceptor-generated cAMP in brown fat cells. AB - The relationship between cAMP levels and thermogenesis was investigated in brown fat cells from Syrian hamsters. Irrespective of whether the selective beta3-, beta2-, and beta1-agonists BRL 37344, salbutamol, and dobutamine or the physiological agonist norepinephrine was used to stimulate the cells, increases in cAMP levels were mediated via the beta3-receptor, as were the thermogenic effects. However, the relationship "thermogenesis per cAMP" was much lower for agents other than norepinephrine. Similarly, forskolin, although more potent than norepinephrine in elevating cAMP, was less potent in inducing thermogenesis. The selective alpha1-agonist cirazoline was in itself without effect on cAMP levels or thermogenesis, but when added to forskolin-stimulated cells, potentiated thermogenesis, up to the norepinephrine level, without affecting cAMP. This potentiation could not be inhibited by chelerythrine, but could be mimicked by Ca2+ ionophores. It was apparently not mediated via calmodulin-dependent protein kinase and was not an effect on mitochondrial respiratory control. The ability of all cAMP-elevating agents to induce thermogenesis in brown fat cells has earlier been interpreted to mean that it is only through the beta-receptors and the resulting increase in cAMP levels that thermogenesis is induced. However, it is here concluded that the thermogenic response to norepinephrine involves two interacting parts, one mediated via beta-receptors and cAMP and the other via alpha1-receptors and increases in cytosolic Ca2+ levels. PMID- 9407063 TI - Characterization and mechanism of action of Drosophila ribosomal protein S3 DNA glycosylase activity for the removal of oxidatively damaged DNA bases. AB - We recently demonstrated that Drosophila ribosomal protein S3 specifically cleaved duplex oligodeoxynucleotides at sites of 7,8-dihydro-8-oxoguanine (8 oxoGua), presumably due to S3 protein possessing an N-glycosylase activity that is associated with its known apurinic/apyrimidinic (AP) lyase activity. Here we show, using DNA substrates prepared by gamma-irradiation under N2O and analyzed by gas chromatography/isotope-dilution mass spectrometry, that S3 protein efficiently liberates 8-oxoGua as a free base from the damaged DNA substrate. Of the 15 additional modified bases present in the DNA substrate, the only other one acted on by S3 protein was 2,6-diamino-4-hydroxy-5-formamidopyrimidine (FapyGua). Specificity constants measured for the removal of 8-oxoGua and FapyGua indicate that S3 protein has a similar preference for both of these modified purines. Having established that S3 protein contains an N-glycosylase activity, we next examined the repair of duplex oligonucleotides containing 8-oxoGua (8-oxoGua-37 mer) positioned opposite Cyt, Gua, Thy, or Ade. Significant cleavage of the 8 oxoGua-37-mer was only detected for an opposing Cyt. Moreover, we show that an imino covalent enzyme-substrate intermediate is formed between S3 protein and 8 oxoGua-37-mer, a result similar to other DNA repair enzymes that catalyze N glycosylase/AP lyase-type reactions at sites of DNA damage. PMID- 9407064 TI - Phosphatidylcholine-specific phospholipase C regulates thapsigargin-induced calcium influx in human lymphocytes. AB - The involvement of phosphatidylcholine-specific phospholipase C (PC-PLC) and D (PC-PLD) in the regulation of the thapsigargin-induced Ca2+ increase was investigated. Pretreatment of human lymphocytes with the PC-PLC inhibitors D609 or U73122 enhanced the thapsigargin-induced Ca2+ influx. By contrast, no effect was observed in the presence of phospholipase D inhibitor butanol. Addition of exogenous PC-PLC but not PC-PLD to lymphocytes prestimulated with thapsigargin led to a decrease of intracellular Ca2+. In addition, thapsigargin was shown to release diacylglycerol (DAG) from cellular phosphatidylcholine pools. The thapsigargin-induced DAG formation was inhibited by U73122 and D609 but not by butanol. Moreover, no formation of the PC-PLD activity marker phosphatidylbutanol was detected. Thapsigargin-induced DAG formation was dependent on the Ca2+ entry, as it was abolished in the absence of extracellular Ca2+ or in the presence of Ni2+. Further investigations demonstrated that the inhibition of the cellular DAG target, protein kinase C (PKC), enhanced thapsigargin-induced Ca2+ increase, whereas direct PKC activation had an inhibitory effect. Taken together, our results reveal the involvement of PC-PLC in the regulation of the thapsigargin induced Ca2+ increase and point to the existence of a physiologic feedback mechanism activated by Ca2+ influx and acting via consecutive activation of PC PLC and PKC to limit the rise of intracellular Ca2+. PMID- 9407065 TI - The WW domain of neural protein FE65 interacts with proline-rich motifs in Mena, the mammalian homolog of Drosophila enabled. AB - The neural protein FE65 contains two types of protein-protein interaction modules: one WW binding domain and two phosphotyrosine binding domains. The carboxyl-terminal phosphotyrosine binding domain of FE65 interacts in vivo with the beta-amyloid precursor protein, which is implicated in Alzheimer disease. To understand the function of this adapter protein, we identified binding partners for the FE65 WW domain. Proline-rich sequences sharing a proline-proline-leucine proline core motif were recovered by screening expression libraries for ligands of the FE65 WW domain. Five proteins of molecular masses 60, 75, 80, 140, and 200 kDa could be purified from mouse brain lysates by affinity to the FE65 WW domain. We identified two of these five proteins as the 80- and 140-kDa isoforms encoded by Mena, the mammalian homolog of the Drosophila Enabled gene. Using the SPOTs technique of peptide synthesis, we identified the sequences in Mena that interact with the FE65 WW domain and found that they contain the signature proline-proline leucine-proline motif. Finally, we demonstrated that Mena binds to FE65 in vivo by coimmunoprecipitation assay from COS cell extracts. The specificity of the Mena-FE65 WW domain association was confirmed by competition assays. Further characterization of the FE65-Mena complex may identify a physiological role for these proteins in beta-amyloid precursor protein biogenesis and may help in understanding the mechanism of molecular changes that underlie Alzheimer disease. PMID- 9407067 TI - Mapping and characterization of the functional domains responsible for the differential activity of the A and B isoforms of the human progesterone receptor. AB - In humans, the biological response to progesterone is mediated by two distinct forms of the progesterone receptor (human (h) PR-A, 94 kDa and hPR-B, 114 kDa). These two isoforms are transcribed from distinct estrogen-inducible promoters within a single copy PR gene; the only difference between them is that the first 164 amino acids of hPR-B (B-upstream sequence) are absent in hPR-A. In most cell lines such as MCF-7 (human breast cancer cells), CV-1 (monkey kidney fibroblasts), and HeLa (human cervical carcinoma cells), hPR-A functions as a transcriptional repressor, whereas hPR-B functions as a transcriptional activator of progesterone-responsive genes. Interestingly, in these cell contexts, hPR-A also acts as a trans-dominant repressor of the transcriptional activity of other steroid hormone receptors. In contrast to hPR-A, which functions predominantly as a ligand-dependent transcriptional repressor, we show in this study that the A isoform of the chicken PR (cPR-A) lacks this trans-dominant repressor function and is a transcriptional activator in all contexts examined. By constructing chimeras between the N-terminal domains of the chicken and human PR, we mapped the trans-dominant repressor function of hPR-A to the first 140 amino acids of the protein. Notably, when this 140-amino acid "repressor" domain is placed onto chicken PR-A, the activity of the latter changes from a transcriptional activator to a repressor. Interestingly, however, this "repressor domain" is necessary, but not sufficient, for trans-repression as it is inactive when it is tethered to a heterologous protein. This suggests that the trans-repression function is comprised not only of the repressor domain of hPR-A but also requires the context of the receptor to function. The identification of a discrete inhibitory region within hPR-A which is transferable to another receptor implies that this region interacts with a set of transcription factors or adaptors that are distinct from those recognized by hPR-B, the identification of which will be required to define the mechanism by which hPR-A modulates steroid hormone receptor transcriptional activity. Thus, although chickens and humans both produce two very similar forms of the progesterone receptor, it is clear from these studies that the mechanism of action of progesterone in these two systems is quite different. PMID- 9407066 TI - Cysteine and disulfide scanning reveals a regulatory alpha-helix in the cytoplasmic domain of the aspartate receptor. AB - The transmembrane, homodimeric aspartate receptor of Escherichia coli and Salmonella typhimurium controls the chemotactic response to aspartate, an attractant, by regulating the activity of a cytoplasmic histidine kinase. The cytoplasmic domain of the receptor plays a central role in both kinase regulation and sensory adaptation, although its structure and regulatory mechanisms are unknown. The present study utilizes cysteine and disulfide scanning to probe residues Leu-250 through Gln-309, a region that contains the first of two adaptive methylation segments within the cytoplasmic domain. Following the introduction of consecutive cysteine residues by scanning mutagenesis, the measurement of sulfhydryl chemical reactivities reveals an alpha-helical pattern of exposed and buried positions spanning residues 270-309. This detected helix, termed the "first methylation helix," is strongly amphiphilic; its exposed face is highly anionic and possesses three methylation sites, while its buried face is hydrophobic. In vivo and in vitro assays of receptor function indicate that inhibitory cysteine substitutions are most prevalent on the buried face of the first methylation helix, demonstrating that this face is involved in a critical packing interaction. The buried face is further analyzed by disulfide scanning, which reveals three "lock-on" disulfides that covalently trap the receptor in its kinase-activating state. Each of the lock-on disulfides cross-links the buried faces of the two symmetric first methylation helices of the dimer, placing these helices in direct contact at the subunit interface. Comparative sequence analysis of 56 related receptors suggests that the identified helix is a conserved feature of this large receptor family, wherein it is likely to play a general role in adaptation and kinase regulation. Interestingly, the rapid rates and promiscuous nature of disulfide formation reactions within the scanned region reveal that the cytoplasmic domain of the full-length, membrane-bound receptor has a highly dynamic structure. Overall, the results demonstrate that cysteine and disulfide scanning can identify secondary structure elements and functionally important packing interfaces, even in proteins that are inaccessible to other structural methods. PMID- 9407068 TI - The cytoplasmic juxtamembrane domain of the epidermal growth factor receptor contains a novel autonomous basolateral sorting determinant. AB - The epidermal growth factor receptor (EGFR) is localized at the basolateral membrane of most epithelial cells in vivo and in cell lines used to study membrane protein sorting. The goal of this study was to define the molecular basis of polar EGFR membrane expression using the Madin-Darby canine kidney cell model. We have identified a 23-amino acid segment located near the cytoplasmic face of the membrane spanning domain (residues Lys-652 to Ala-674) that is necessary and sufficient for targeting EGFRs from the trans-Golgi network directly to the basolateral plasma membrane. Furthermore, the sequence between residues Lys-652 and Ala-674 is sufficient to direct the extracellular domain of an apical membrane protein, decay accelerating factor, to the basolateral membrane. In the absence of this cytoplasmic basolateral sorting signal, information within the extracellular ligand binding domain is sufficient to target EGFRs from the trans-Golgi network directly to the apical plasma membrane. The EGFR basolateral sorting determinant does not have sequence and structural requirements common to most basolateral membrane proteins and does not overlap any of the known EGFR endocytic signals. This 23-residue sequence lies in a predicted amphipathic helical structure, leading us to postulate that hydrophobic and/or electrostatic interactions may be important for activity of this autonomous basolateral sorting determinant. PMID- 9407069 TI - Differential regulation of interleukin-8 and intercellular adhesion molecule-1 by H2O2 and tumor necrosis factor-alpha in endothelial and epithelial cells. AB - The reactive oxygen intermediate H2O2 can function as a signaling molecule to activate gene expression. In this study, we demonstrate that oxidant stress induced by tumor necrosis factor alpha (TNFalpha) or H2O2 differentially regulates intercellular adhesion molecule-1 (ICAM-1) and interleukin-8 (IL-8) gene expression in endothelial and epithelial cells. Northern blot analysis revealed that TNFalpha induced both ICAM-1 and IL-8 expression in either the A549 lung epithelial cell line or the human microvessel endothelial cell line (HMEC 1). In contrast, H2O2 selectively induced only ICAM-1 in HMEC-1 and only IL-8 in A549. This cell type-specific pattern of IL-8 expression was also observed in several other endothelial and epithelial cells. TNFalpha induced greater IL-8 gene expression as compared with H2O2, but the kinetics of induction were similar. The induction of epithelial IL-8 message was accompanied by a corresponding increase in functional IL-8 protein secretion as determined by a neutrophil motility assay. The increased neutrophil motility stimulated by conditioned media from H2O2- or TNFalpha-exposed A549 cells was completely inhibited by an anti-IL-8 antibody. TNFalpha and H2O2 also induced a differential pattern of CC chemokine expression in A549. While TNFalpha induced both RANTES and MCP-1, H2O2 induced only MCP-1. These data suggest that epithelial cells under oxidant stress contribute to the inflammatory cytokine network by selective production of IL-8, MCP-1, and RANTES, which may critically influence the site specific recruitment of leukocyte subsets. PMID- 9407070 TI - Transcriptional regulation of sodium transport by vasopressin in renal cells. AB - We have examined whether arginine vasopressin (AVP) can induce a long-term modulation of transepithelial ion transport in addition to its well known short term effect. In the RCCD1 rat cortical collecting duct cell line, an increase in both short-circuit current and 22Na transport was observed after several hours of 10(-8) M AVP treatment (a concentration above the in vivo physiological range). This delayed effect was partially prevented by apical addition of 10(-5) M amiloride and was blocked by 10(-6) M actinomycin D and 2 x 10(-6) M cycloheximide. The amounts of mRNA encoding the alpha1 (not beta1) subunit of Na+/K+-ATPase and the beta and gamma (not alpha) subunits of the amiloride sensitive epithelial Na+ channel were significantly increased by AVP treatment. The increase in mRNA was blocked by actinomycin D, not by amiloride, suggesting a Na+-independent increase in the rate of transcription of these subunits. The translation rates of the alpha1 subunit of Na+/K+-ATPase and the beta and gamma subunits of the rat epithelial sodium channel increased significantly, whereas the translation rates of the other subunits remained unchanged. Finally, the number of Na+ channels present in the apical membrane of the cells increased, as demonstrated by enhanced specific [3H]phenamil binding. PMID- 9407072 TI - N-Nmoc-L-glutamate, a new caged glutamate with high chemical stability and low pre-photolysis activity. AB - We report the synthesis, the physicochemical characterization, and biological evaluation of a new caged glutamate, N-(o-nitromandelyl)oxycarbonyl-L-glutamic acid (Nmoc-Glu), that liberates free glutamate on photolysis. The low affinity of certain glutamate receptors and their rapid entry into desensitization have effectively prevented the creation of an ideal caged glutamate. In the absence of an ideal compound, Nmoc-Glu was designed to resist spontaneous hydrolysis while maintaining reasonable photorelease yield and kinetics. Chemical and physiological analyses reveal that Nmoc-Glu, indeed, has exceptionally low residual activity and high chemical stability. The quantum yield of Nmoc-Glu is 0.11. Photolytic uncaging and release of free glutamate occur in two steps, consisting of an initial light-induced cleavage that proceeds on the sub millisecond time scale, and a subsequent light-independent, pH-dependent decarboxylation step that proceeds on the millisecond time scale. The low residual activity and high chemical stability of Nmoc-Glu are important advantages in applications where pre-photolysis Glu receptor activation and desensitization must be minimized. PMID- 9407071 TI - Effects of the inter-ring communication in GroEL structural and functional asymmetry. AB - The chaperonin GroEL consists of a double-ring structure that assists protein folding in the presence of GroES and ATP. Recent studies suggest that the 7-mer ring is the functional unit where protein folding takes place. Nevertheless, both GroEL rings are required to complete the reaction cycle through signals transmitted between the two rings. Electron microscopy, image processing, and biochemical analysis of GroEL, a single-ring mutant (SR1) and a inter-ring communication affected mutant (A126V), in the presence of ATP and adenylyl imidodiphosphate, have allowed the identification of a conformational change in the apical domains that is strictly dependent on the communication between the two GroEL rings. It is deduced from these results that the binding of nucleotide to both GroEL rings generates, as a consequence of the inter-ring communication, a functionally and structurally asymmetric particle. This asymmetric particle has a ring with a small conformational change in its apical domains and high affinity toward unfolded substrate and GroES, and the other ring has a larger conformational change in its apical domains and lower affinity toward substrate and GroES. PMID- 9407073 TI - Identification of amino acids contributing to high and low affinity d tubocurarine sites in the Torpedo nicotinic acetylcholine receptor. AB - d-Tubocurarine (dTC) is a potent competitive antagonist of the Torpedo nicotinic acetylcholine receptor (nAChR) that binds non-equivalently to the two agonist sites (Kd values of 30 nM and 8 microM). When nAChR-rich membranes equilibrated with [3H]dTC are irradiated with 254 nm UV light, [3H]dTC is covalently incorporated into the alpha-, gamma-, and delta-subunits in a concentration dependent and agonist-inhibitable manner, consistent with the localization of the high and low affinity dTC binding sites at the alpha-gamma- and alpha-delta subunit interfaces, respectively (Pedersen, S. E. and Cohen, J. B. (1990) Proc. Natl. Acad. Sci. U. S. A. 87, 2785-2789). We report on the amino acids within alpha-, gamma-, and delta-subunits that are the sites of specific photoincorporation of [3H]dTC. Subunits isolated from nAChR-rich membranes photolabeled with [3H]dTC were subjected to enzymatic digestion, and peptides containing 3H were isolated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and/or reversed-phase high performance liquid chromatography. Isolated peptides were then subjected to NH2-terminal sequence analysis to identify specifically labeled residues. Within the alpha-subunit, 95% of specific incorporation was contained within a 20-kDa proteolytic fragment beginning at Ser 173, with alphaTyr-190 the primary site of [3H]dTC photoincorporation and alphaCys-192 and alphaTyr-198 labeled at lower efficiency. Within gamma- and delta-subunits, specific labeling was contained within proteolytic fragments of 14 and 21 kDa, respectively, beginning at gammaAla-49 and deltaThr-51. gammaTrp 55 and deltaTrp-57 were identified as the sites of specific [3H]dTC photoincorporation. Sequence alignment studies reveal gammaTrp-55 and deltaTrp-57 to be homologous residues at whose position in receptor subunit primary structure a unique pattern of conservation exists in all nAChR (neuronal and muscle). Specifically, all subunits that associate with an alpha-subunit to form an agonist site contain a tryptophan homologous to gammaTrp-55/deltaTrp-57. This pattern of conservation may indicate a functional significance for tryptophan at that location in all nAChR agonist sites. PMID- 9407074 TI - Characterization and localization of P2 receptors in rat submandibular gland acinar and duct cells. AB - [Ca2+]i and the Cl- current were measured in isolated submandibular gland acinar and duct cells to characterize and localize the purinergic receptors expressed in these cells. In both cell types 2'-3'-benzoylbenzoyl (Bz)-ATP and ATP increased [Ca2+]i mainly by activation of Ca2+ influx. UTP had only minimal effect on [Ca2+]i at concentrations between 0.1 and 1 mM. However, a whole cell current recording showed that all nucleotides effectively activated Cl- currents. Inhibition of signal transduction through G proteins by guanyl-5'-beta thiophosphate revealed that the effect of ATP on Cl- current was mediated in part by activation of a G protein-coupled and in part by a G protein-independent receptor. BzATP activated exclusively the G protein-independent portion, whereas UTP activated only the G protein-dependent portion of the Cl- current. Measurement of [Ca2+]i in the microperfused duct showed that ATP stimulated a [Ca2+]i increase when applied to the luminal or the basolateral sides. BzATP increased [Ca2+]i only when applied to the luminal side, whereas UTP at 100 microM increased -Ca2+-i only when applied to the basolateral side. The combined results suggest that duct and possibly acinar cells express P2z receptors in the luminal and P2u receptors in the basolateral membrane. PMID- 9407075 TI - Membrane-specific regulation of Cl- channels by purinergic receptors in rat submandibular gland acinar and duct cells. AB - Measurement of [Cl-]i and the Cl- current in the rat salivary submandibular gland (SMG) acinar and duct cells was used to evaluate the role of Cl- channels in the regulation of [Cl-]i during purinergic stimulation. Under resting conditions [Cl ]i averaged 56 +/- 8 and 26 +/- 7 mM in acinar and duct cells, respectively. In both cells, stimulation with 1 mM ATP resulted in Cl- efflux and subsequent influx. Inhibition of NaKCl2 cotransport had no effect on [Cl-]i changes in duct cells and inhibited only about 50% of Cl- uptake in acinar cells. Accordingly, low levels of expression of NaKCl2 cotransporter protein were found in duct cells. Acinar cells expressed high levels of the cotransporter. Measurement of Cl current under selective conditions revealed that acinar and duct cells express at least five distinct Cl- channels; a ClCO-like, volume-sensitive, inward rectifying, Ca2+-activated and CFTR-like Cl- currents. ATP acting on both cell types activated at least two channels, the Ca2+-activated Cl- channel and a Ca2+ independent glibenclamide-sensitive Cl--current, possibly cystic fibrosis transmembrane regulator (CFTR). Of the many nucleotides tested only 2'-3' benzoylbenzoyl (Bz)-ATP and UTP activated Cl- channels in SMG cells. Despite their relative potency in increasing [Ca2+]i, BzATP in both SMG cell types largely activated the Ca2+-independent, glibenclamide-sensitive Cl- current, whereas UTP activated only the Ca2+-dependent Cl- current. We interpret this to suggest that BzATP and UTP largely activate Cl- channels residing in the membrane expressing the receptor for the active nucleotide. The present studies reveal a potentially new mechanism for transcellular Cl- transport in a CFTR-expressing tissue, the SMG. Coordinated action of the P2z (luminal) and P2u (basolateral) receptors can mediate part of the transcellular Cl- transport by acinar and duct cells to determine the final electrolyte composition of salivary fluid. PMID- 9407077 TI - Properties and phosphorylation sites of baculovirus-expressed nuclear inhibitor of protein phosphatase-1 (NIPP-1). AB - NIPP-1 is the RNA-binding subunit of a major species of protein phosphatase-1 in the nucleus. We have expressed nuclear inhibitor of protein phosphatase-1 (NIPP 1) in Sf9 cells, using the baculovirus-expression system. The purified recombinant protein was a potent (Ki = 9.9 +/- 0.3 pM) and specific inhibitor of protein phosphatase-1 and was stoichiometrically phosphorylated by protein kinases A and CK2. At physiological ionic strength, phosphorylation by these protein kinases drastically decreased the inhibitory potency of free NIPP-1. Phosphorylation of NIPP-1 in a heterodimeric complex with the catalytic subunit of protein phosphatase-1 resulted in an activation of the holoenzyme without a release of NIPP-1. Sequencing and phosphoamino acid analysis of tryptic phosphopeptides enabled us to identify Ser178 and Ser199 as the phosphorylation sites of protein kinase A, whereas Thr161 and Ser204 were phosphorylated by protein kinase CK2. These residues all conform to consensus recognition sites for phosphorylation by protein kinases A or CK2 and are clustered near a RVXF sequence that has been identified as a motif that interacts with the catalytic subunit of protein phosphatase-1. PMID- 9407076 TI - Ras-stimulated extracellular signal-related kinase 1 and RhoA activities coordinate platelet-derived growth factor-induced G1 progression through the independent regulation of cyclin D1 and p27. AB - Platelet-derived growth factor (PDGF)-induced Ras activation is required for G1 progression in Chinese hamster embryo fibroblasts (IIC9 cells). Ras stimulates both extracellular signal-related kinase (ERK) activation and RhoA activation in response to PDGF stimulation. Inhibition of either of these Ras-stimulated pathways results in growth arrest. We have shown previously that Ras-stimulated ERK activation is essential for the induction and continued G1 expression of cyclin D1. In this study we examine the role of Ras-induced RhoA activity in G1 progression. Unstimulated IIC9 cells expressed high levels of the G1 cyclin dependent kinase inhibitor p27(KIP1). Stimulation with PDGF resulted in a dramatic decrease in p27(KIP1) protein expression. This decrease was attributed to increased p27(KIP1) protein degradation. Overexpression of dominant-negative forms of Ras or RhoA completely blocked PDGF-induced p27(KIP1) degradation, but only dominant-negative Ras inhibited cyclin D1 protein expression. C3 transferase also inhibited PDGF-induced p27(KIP1) degradation, thus further implicating RhoA in p27(KIP1) regulation. Overexpression of dominant-negative ERK resulted in inhibition of PDGF-induced cyclin D1 expression but had no effect on PDGF-induced p27(KIP1) degradation. These data suggest that Ras coordinates the independent regulation of cyclin D1 and p27(KIP1) expression by the respective activation of ERK and RhoA and that these pathways converge to determine the activation state of complexes of cyclin D1 and cyclin-dependent kinase in response to mitogen. PMID- 9407078 TI - Agonist-independent activation of Gz by the 5-hydroxytryptamine1A receptor co expressed in Spodoptera frugiperda cells. Distinguishing inverse agonists from neutral antagonists. AB - The human 5-hydroxytryptamine1A receptor, when expressed in Spodoptera frugiperda (Sf9) cells, facilitates the binding of [35S]GTPgammaS to a co-expressed GTP binding regulatory protein, Gz, consistent with constitutive activity. The antagonists 4-(2'-methoxyphenyl)-1-[2'(n-2"-pyridinyl)-p-iodobenzamido]ethyl-p ipe razine (p-MPPI) and the related fluorobenzamido analogue p-MPPF had little (p MPPI) or no (p-MPPF) effect on this activity. In contrast, a third antagonist, the neuroleptic spiperone, produced an almost complete suppression. Thus, using G protein activation as an index of receptor activity, p-MPPF was classified as a neutral antagonist, p-MPPI as a partial inverse agonist, and spiperone as essentially a full inverse agonist. As predicted, spiperone displayed properties consistent with a special form of noncompetitive antagonism when used to displace the agonist [125I]R-(+)-trans-8-hydroxy-2-[N-n-propyl-N-(3'-iodo-2'-propenyl)amin o]tetralin. Our data profile Sf9 cells as a unique vehicle for the characterization of inverse agonists, as these cells support a systematic pairing of mammalian receptors and G proteins, quantitative assays of G protein activation, and unambiguously labeled populations of coupled and uncoupled receptors. PMID- 9407079 TI - A 21-kDa C-terminal fragment of protein-disulfide isomerase has isomerase, chaperone, and anti-chaperone activities. AB - A catalyst of disulfide formation and isomerization during protein folding, protein-disulfide isomerase (PDI) has two catalytic sites housed in two domains homologous to thioredoxin, one near the N terminus and the other near the C terminus. The thioredoxin domains, by themselves, can catalyze disulfide formation, but they are unable to catalyze disulfide isomerizations (Darby, N. J. and Creighton, T. E. (1995) Biochemistry 34, 11725-11735). A 21-kDa, C-terminal fragment of PDI (amino acids 308-491), termed weePDI, comprises the C-terminal third of the molecule. The kcat for ribonuclease oxidative folding by weePDI is 0.26 +/- 0.02 min-1, 3-fold lower than the wild-type enzyme but indistinguishable from the activity of a full-length mutant of PDI in which both active site cysteines of the N-terminal thioredoxin domain have been mutated to serine. Eliminating the ability of weePDI to escape easily from covalent complexes with substrate by mutating the active site cysteine nearer the C terminus to serine has a large effect on the isomerase activity of weePDI compared with its effect on the full-length enzyme. weePDI also displays chaperone and anti-chaperone activity characteristic of the full-length molecule. As isolated, weePDI is a disulfide-linked dimer in which the single cysteine (Cys-326) outside active site cross-links two weePDI monomers. The presence of the intermolecular disulfide decreases the activity by more than 2-fold. The results imply that the functions of the core thioredoxin domains of PDI and other members of the thioredoxin superfamily might be modified quite easily by the addition of relatively small accessory domains. PMID- 9407080 TI - Disulfide bond assignment in human interleukin-7 by matrix-assisted laser desorption/ionization mass spectroscopy and site-directed cysteine to serine mutational analysis. AB - Interleukin-7 (IL-7) is a proteinaceous biological response modifier that has a bioactive tertiary structure dependent on disulfide bond formation. Disulfide bond assignments in human (h)IL-7 are based upon the results of matrix-assisted laser desorption/ionization (MALDI) mass spectroscopy and Cys to Ser mutational analyses. A gene encoding the hIL-7 was synthesized incorporating Escherichia coli codon usage bias and was used to express biologically active protein as determined by stimulation of precursor B-cell proliferation. MALDI mass spectroscopic analysis of trypsin-digested hIL-7 was performed and compared with the anticipated results of a simulated tryptic digestion. Many of the anticipated hIL-7 tryptic fragments were detected including one with a molecular mass equivalent to the sum of two polypeptides linked through a disulfide bond formed from Cys residues (Cys3 and Cys142). Subsequently, Cys to Ser substitution mutational analyses were performed. A hIL-7 variant with all six Cys substituted with Ser was found to be biologically inactive (EC50 > 1 x 10(-7) M). In contrast, a family of single disulfide bond-forming variants of hIL-7 were constructed by reintroducing Cys pairs (Cys3-Cys142, Cys35-Cys130, and Cys48 Cys93), and each could stimulate cell proliferation with an EC50 of 4 x 10(-9), 2 x 10(-8), and 2 x 10(-9) M, respectively. In single disulfide bond-forming mutants of hIL-7, the ability to stimulate cell proliferation was abolished in the presence of 2 mM dithiothreitol. The results presented strongly suggest that only a single disulfide bond is required for hIL-7 to form a tertiary structure capable of stimulating precursor B-cell proliferation. PMID- 9407082 TI - Substrate recognition by the leucyl/phenylalanyl-tRNA-protein transferase. Conservation within the enzyme family and localization to the trypsin-resistant domain. AB - The leucyl/phenylalanyl-tRNA-protein transferase (L/F-transferase) from Escherichia coli catalyzes a peptidyltransferase reaction that results in the N terminal aminoacylation of acceptor proteins using Leu-, Phe-, and Met-tRNAs as amino acid donors. We demonstrated that L/F-transferase homologs are widely distributed throughout the eubacteria, supporting our proposal that the enzyme family is ancient and catalyzes early peptide bond synthesis. However, here we present data suggesting that the L/F-transferase is not a homolog of the peptidyltransferase enzymes involved in cell wall peptidoglycan biosynthesis in Gram-positive species, such as Staphylococcus aureus. A sequence comparison of the known L/F-transferase homologs began to identify the essential residues required to catalyze a peptidyltransferase reaction and revealed that <20% of the residues were invariant within the L/F-transferase family. Despite this sequence variation, substrate specificity was broadly conserved, and L/F-transferase homologs from Providencia stuartii, Vibrio cholerae, Neisseria gonorrhoeae, and the cyanobacterium Synechocystis sp. all complemented an E. coli aat mutant (lacking L/F-transferase activity) for the degradation of N-end rule substrates. In vitro comparison of the most divergent L/F-transferase homologs, from E. coli and the cyanobacterium Synechocystis sp., revealed near-complete conservation of both substrate specificity and secondary structure. Finally, we demonstrated that variants of the E. coli L/F-transferase, lacking either 33 or 78 N-terminal residues, retained measurable peptidyltransferase activity and wild type substrate specificity. Overall, our results identified an essential core of an L/F-transferase and revealed that a peptidyltransferase catalyst may be constructed from approximately 120 amino acids. PMID- 9407081 TI - Different domains of mammalian ADP-ribosylation factor 1 mediate interaction with selected target proteins. AB - Mammalian ADP-ribosylation factor 1 (mARF1) is a small GTP-binding protein that is activated by a Golgi guanine nucleotide exchange factor. Once bound to the Golgi membranes in the GTP form, mARF1 initiates the recruitment of the adaptor protein 1 (AP-1) complex and coatomer (COPI) onto these membranes and activates phospholipase D1 (PLD1). To map the domains of mARF1 that are important for these activities, we constructed chimeras between mARF1 and Saccharomyces cerevisiae ARF2, which functions poorly in all of these processes except COPI recruitment. The carboxyl half of mARF1 (amino acids 95-181) was essential for activation by the Golgi guanine nucleotide exchange factor, whereas a separate domain (residues 35-94) was required to effectively activate PLD1 and to promote efficient AP-1 recruitment. Since residues 35-94 of mARF1 are critical for optimal activity in both PLD1 activation and AP-1 recruitment, we hypothesize that this region of ARF contains residues that interact with effector molecules. PMID- 9407083 TI - Three-dimensional structure of L-2-haloacid dehalogenase from Xanthobacter autotrophicus GJ10 complexed with the substrate-analogue formate. AB - The L-2-haloacid dehalogenase from the 1,2-dichloroethane degrading bacterium Xanthobacter autotrophicus GJ10 catalyzes the hydrolytic dehalogenation of small L-2-haloalkanoic acids to yield the corresponding D-2-hydroxyalkanoic acids. Its crystal structure was solved by the method of multiple isomorphous replacement with incorporation of anomalous scattering information and solvent flattening, and was refined at 1.95-A resolution to an R factor of 21.3%. The three dimensional structure is similar to that of the homologous L-2-haloacid dehalogenase from Pseudomonas sp. YL (1), but the X. autotrophicus enzyme has an extra dimerization domain, an active site cavity that is completely shielded from the solvent, and a different orientation of several catalytically important amino acid residues. Moreover, under the conditions used, a formate ion is bound in the active site. The position of this substrate-analogue provides valuable information on the reaction mechanism and explains the limited substrate specificity of the Xanthobacter L-2-haloacid dehalogenase. PMID- 9407084 TI - SNAP-23 is not cleaved by botulinum neurotoxin E and can replace SNAP-25 in the process of insulin secretion. AB - The synaptosomal-associated protein of 25 kDa (SNAP-25) is expressed in neurons and endocrine cells. It has been shown to play an important role in the release mechanism of neurotransmitters and peptide hormones, including insulin. Thus, when insulin-secreting cells are permeabilized and treated with botulinum neurotoxin E (BoNT/E), SNAP-25 is hydrolyzed, and insulin secretion is inhibited. Recently SNAP-23, a more generally expressed isoform of SNAP-25, has been described. The functional role of SNAP-23 has not been investigated to date. It is now shown that SNAP-23 is resistant to cleavage by BoNT/E. It was therefore possible to test whether transfection of HIT (transformed pancreatic B-) cells with SNAP-23 reconstitutes insulin release from BoNT/E treated cells, in which SNAP-25 is inactivated by the toxin. The results show that SNAP-23 is able to replace SNAP-25 when it is overexpressed. While these results demonstrate that SNAP-23 is a functional homologue of SNAP-25, able to function in regulated exocytosis, they indicate that SNAP-23 may be inefficient in this process. This suggests that both isoforms may have their own specific binding partners and discrete, albeit mechanistically similar, functional roles within the cell. PMID- 9407085 TI - The testis-specific histone H1t gene is strongly repressed by a G/C-rich region just downstream of the TATA Box. AB - H1t is a testis-specific histone 1 variant restricted to the male germ line and expressed only in pachytene spermatocytes. Understanding the regulation of the H1t gene is an interesting challenge as its promoter shares all of the recognized control elements of standard somatic H1 genes, yet H1t is not expressed in somatic or in early spermatogenic cells. To investigate the mechanism of this apparent repression, we exchanged three promoter subregions between H1t and a major somatic H1 gene (H1d) by introduction of suitable restriction sites just 5' of the TATA box and 3' of the conserved H1 AC box. Hybrid promoters were joined to a lacZ reporter gene and assayed by transient transfection in NIH3T3 fibroblasts. In this system the wild type H1d promoter was 20-fold stronger than the H1t promoter. Much of this difference in activity was traced to inhibitory sequences immediately downstream of the TATA box in H1t, although sequences upstream of the H1t AC box and within the H1t 5'-untranslated region played some role as well. A series of deletions and short oligonucleotide mutations scanned across the region between the TATA box and cap site identified two tracts of C (GC box 2) as the inhibitory sequences. While both Sp1 and Sp3 bind to this region weakly in vitro, they are unlikely to be responsible for the inhibitory effect of GC box 2, and additional binding proteins (CTB-4 and CTB-5) were identified by electrophoretic mobility shift assays as better candidates for mediating the repressive effect. When repression of the H1t promoter was relieved by mutation of GC box 2, additional mutations introduced into GC box 1 upstream of the CAAT box led to a large decrease in activity, indicating that these two G/C-rich elements have opposite effects on promoter activity. PMID- 9407086 TI - Functional importance of platelet-derived growth factor (PDGF) receptor extracellular immunoglobulin-like domains. Identification of PDGF binding site and neutralizing monoclonal antibodies. AB - The biological effects of platelet-derived growth factor (PDGF) are mediated by alpha- and beta-PDGF receptors (PDGFR), which have an intracellular tyrosine kinase domain and an extracellular region comprising five immunoglobulin-like domains (D1-D5). Using deletion mutagenesis we mapped the PDGF binding site in each PDGFR to the D2-D3 region. In the case of alpha-PDGFR, 125I-PDGF AA and 125I PDGF BB bound to the full-length extracellular domain, D1-D5, and D2-D3 with equal affinity (Kd = 0.21-0.42 nM). Identical results were obtained for 125I-PDGF BB binding to beta-PDGFR mutants D1-D5 and D2-D3, establishing that D1, D4, and D5 do not contribute to PDGF binding. Monoclonal antibodies (mAb) directed against individual PDGFR Ig-like domains were used to extend these observations. The anti-D1 mAb 1E10E2 and anti-D5 mAb 2D4G10 had no effect on alpha- or beta PDGFR function, respectively. In contrast, mAb 2H7C5 and 2A1E2 directed against D2 of the alpha- and beta-receptor, respectively, blocked PDGF binding, receptor autophosphorylation and mitogenic signaling with IC50 values of 0.1-3.0 nM. An anti-D4 mAb 1C7D5 blocked beta-receptor autophosphorylation and signaling without inhibiting PDGF binding consistent with the observation that D4 is essential for PDGFR dimerization (Omura, T., Heldin, C.-H., and Ostman, A. (1997) J. Biol. Chem. 272, 12676-12682). mAbs identified here act as potent PDGFR antagonists that can be used as research tools and potentially as therapeutic agents for the treatment of diseases involving unwanted PDGFR signaling. PMID- 9407087 TI - Cloning and expression of acetylcholinesterase from Electrophorus. Splicing pattern of the 3' exons in vivo and in transfected mammalian cells. AB - We cloned and expressed a cDNA encoding acetylcholinesterase (AChE) of type T from Electrophorus electricus organs. When expressed in COS, HEK, and Chinese hamster ovary cells, the AChET subunits generated dimers and tetramers. The cells produced more activity at 27 than at 37 degrees C. The kinetic parameters of a recombinant enzyme, produced in the yeast Pichia pastoris, were close to those of the natural AChE. Analysis of genomic clones showed that the coding sequence is interrupted by an intron that does not exist in Torpedo and differs in its location from that observed in the mouse. This intron is preceded by a sequence encoding a non-conserved 29-amino acid peptide, which does not exist in Torpedo or mammalian AChEs. According to a three-dimensional model, this non-conserved peptide is located at the surface of the protein, opposite from the entry of the catalytic gorge; its deletion did not modify the catalytic parameters. Sequence analyses and expression of various constructs showed that the gene does not contain any H exon. We also found that splicing of transcripts in mammalian cells reveals cryptic donor sites in exons and acceptor sites in introns, which do not appear to be used in vivo. PMID- 9407088 TI - Ionizing radiation-inducible apoptosis in the absence of p53 linked to transcription factor EGR-1. AB - The tumor suppressor protein p53 is a pivotal regulator of apoptosis, and prostate cancer cells that lack p53 protein are moderately resistant to apoptotic death by ionizing radiation. Genes encoding the transcription factor early growth response-1 (EGR-1) and cytokine tumor necrosis factor-alpha (TNF-alpha) were induced upon irradiation of prostate cancer cells, and inhibition of EGR-1 function resulted in abrogation of both TNF-alpha induction and apoptosis. Induction of the TNF-alpha gene by ionizing radiation and EGR-1 was mediated via a GC-rich EGR-1-binding motif in the TNF-alpha promoter. Because TNF-alpha induces apoptosis in prostate cancer cells, these findings suggest that, in the absence of p53, ionizing radiation-inducible apoptosis is mediated by EGR-1 via TNF-alpha transactivation. PMID- 9407089 TI - Neuroserpin, a brain-associated inhibitor of tissue plasminogen activator is localized primarily in neurons. Implications for the regulation of motor learning and neuronal survival. AB - A cDNA clone for the serine proteinase inhibitor (serpin), neuroserpin, was isolated from a human whole brain cDNA library, and recombinant protein was expressed in insect cells. The purified protein is an efficient inhibitor of tissue type plasminogen activator (tPA), having an apparent second-order rate constant of 6. 2 x 10(5) M-1 s-1 for the two-chain form. However, unlike other known plasminogen activator inhibitors, neuroserpin is a more effective inactivator of tPA than of urokinase-type plasminogen activator. Neuroserpin also effectively inhibited trypsin and nerve growth factor-gamma but reacted only slowly with plasmin and thrombin. Northern blot analysis showed a 1.8 kilobase messenger RNA expressed predominantly in adult human brain and spinal cord, and immunohistochemical studies of normal mouse tissue detected strong staining primarily in neuronal cells with occasionally positive microglial cells. Staining was most prominent in the ependymal cells of the choroid plexus, Purkinje cells of the cerebellum, select neurons of the hypothalamus and hippocampus, and in the myelinated axons of the commissura. Expression of tPA within these regions is reported to be high and has previously been correlated with both motor learning and neuronal survival. Taken together, these data suggest that neuroserpin is likely to be a critical regulator of tPA activity in the central nervous system, and as such may play an important role in neuronal plasticity and/or maintenance. PMID- 9407090 TI - Structure and localization of the human gene encoding SR-BI/CLA-1. Evidence for transcriptional control by steroidogenic factor 1. AB - The scavenger receptor, class B, type 1 receptor (SR-BI) mediates the selective transport of lipids from high density lipoprotein to cells. We describe the structure and subchromosomal location of human SR-BI and provide evidence that it is regulated by the transcription factor, steroidogenic factor 1 (SF-1). SR-BI resides on chromosome 12q24.2-qter, spans approximately 75 kilobase pairs, and contains 13 exons. RNA blot analysis of human tissues reveals an expression pattern similar to that described previously for rodents with the highest levels of mRNA in the adrenal gland, ovary, and liver. Unlike rodents, human SR-BI was expressed at high levels in the placenta. The transcription start site for SR-BI was mapped, and DNA sequence analysis revealed a binding site for SF-1 in the proximal 5'-flanking sequence. SF-1, an orphan member of the nuclear hormone receptor gene family, plays a key role in the regulation of steroidogenesis and is expressed at high levels in steroidogenic tissues. SF-1 binds to the SR-BI promoter in a sequence-specific manner, and efficient transcription from this promoter in adrenocortical Y1 cells is dependent on an intact SF-1 site. These data extend our understanding of SF-1 function within steroidogenic tissues and suggest that SR-BI, which serves to supply selected tissues with lipoprotein derived lipids, is part of the repertoire of SF-1-responsive genes involved in steroidogenesis. PMID- 9407091 TI - Structure and oxygen affinity of crystalline des-his-146beta human hemoglobin in the T state. AB - To correlate directly structure with function, the oxygen affinity and the three dimensional structure of crystals of the T quaternary state of des-His-146beta human hemoglobin have been determined by polarized absorption microspectrophotometry and x-ray diffraction crystallography. In des-His-146beta, the COOH-terminal histidine residues of the beta chains of hemoglobin A have been removed. Oxygen binding to crystalline des-His hemoglobin is non-cooperative and independent of pH. The oxygen affinity is 1.7-fold greater than that of the crystalline state of hemoglobin A. Removal of His-146beta results in a small movement of the truncated COOH-terminal peptide and in a very small change in quaternary structure. Previously, similar studies on T state crystals of des-Arg 141alpha hemoglobin showed that removal of the COOH termini of the alpha chains results in much larger effects on oxygen affinity and on quaternary structure. Kinetic studies in solution reveal that at pH 7.0, the rates of CO combination with deoxygenated des-His-146beta in the absence and presence of inositol hexaphosphate are 2.5- and 1.3-fold, respectively, more rapid than for hemoglobin A. The values for des-Arg are 7.6- and 3.9-fold. The properties of the T state of hemoglobin both in the crystal and in solution are influenced to a greater degree by the interactions associated with Arg-141alpha than those associated with His 146beta. PMID- 9407093 TI - Molecular cloning and characterization of a novel cytoplasmic protein-tyrosine phosphatase that is specifically expressed in spermatocytes. AB - We identified a novel gene encoding protein-tyrosine phosphatase using a polymerase chain reaction-based method. Northern blot hybridization of RNAs from various tissues with the polymerase chain reaction-amplified DNA fragment showed that this gene was expressed exclusively in the testis. Complementary DNAs for this gene, termed typ (testis-specific tyrosine phosphatase), were obtained from a mouse testis cDNA library. Nucleotide sequencing of the cDNAs revealed an open reading frame that encoded 426 amino acids. The predicted Typ protein contained a single catalytic domain at the carboxyl-terminal half. No hydrophobic stretch for a possible transmembrane sequence or signal sequence was found, suggesting that Typ is a cytoplasmic protein-tyrosine phosphatase. The amino-terminal half of Typ did not share significant homologies with the other known proteins but contained a region rich in PEST residues. Indirect immunofluorescence studies and in situ hybridization analysis showed that Typ was specifically expressed in testicular germ cells that underwent meiosis. Developmentally, Typ was detected between 2 and 3 weeks after birth, in parallel with the onset of meiosis. Thus, Typ is a new member of the cytoplasmic protein-tyrosine phosphatases that may play an important role(s) in spermatogenesis and/or meiosis. PMID- 9407092 TI - A second determinant of binding to the p75 neurotrophin receptor revealed by alanine-scanning mutagenesis of a conserved loop in nerve growth factor. AB - In the neurotrophin family, variable regions contain solvent-accessible residues important for receptor binding specificity, whereas many of the conserved residues are buried in hydrophobic cores or in the dimer interface. A stretch of six amino acids (from Asp-72 to Asn-77) in nerve growth factor (NGF) represents an exception to this general rule. These residues are highly conserved and yet form an exposed hydrophilic loop region away from other known determinants of receptor binding. We have investigated the functional importance of this region in NGF using alanine-scanning mutagenesis. Individual mutation of Asp-72, Lys-74, or His-75 to alanine (mutants D72A, K74A, and H75A, respectively) reduced the binding affinity for the p75 neurotrophin receptor by 4-10-fold. Only the D72A mutant showed an additional impairment in binding to the TrkA receptor, which was accompanied by reduced biological activity in PC12 cells, indicating a structural and/or conformational effect of this mutation. Replacement of Ser-73 or Asn-77 with alanine (mutants S73A and N77A, respectively) had no measurable effects on receptor binding. The triple mutant K74A/H75A/N77A exhibited properties that were consistent with the combined effects of the individual mutations, namely impaired binding to p75 without deficits in its interaction with TrkA. In contrast, in the triple mutant D72A/S73A/K74A, the simultaneous replacement of Asp-72 and Lys-74 with alanine had a compensatory effect such that binding to both p75 and TrkA was comparable to that of wild-type NGF, despite the deficits seen in the individual replacements. This molecule, however, was produced at low levels, and its biological activity in sympathetic ganglion explants was reduced, which, together with results from TrkA phosphorylation assays, indicated a reduced stability during prolonged culture conditions. Taken together, these data reveal a second region of interaction with the p75 receptor in NGF with the positively charged residues Lys-74 and His-75 as candidate points of contact. In addition, Asp-72 appears to be a structurally important side chain for stabilizing the conformation of the loop through interactions with neighboring residues. PMID- 9407094 TI - Functional coupling of a human retinal metabotropic glutamate receptor (hmGluR6) to bovine rod transducin and rat Go in an in vitro reconstitution system. AB - The cDNA encoding hmGluR6, appended with a 15-amino acid antibody epitope (1D4), was transiently transfected in COS-7 cells. The receptor was purified from COS cell membranes using an antibody affinity column. The purified receptor was then reconstituted into lipid vesicles, and its ability to activate either transducin, the rod photoreceptor-specific GTP-binding protein, or the alpha subunit of Go was assayed in vitro using a guanosine 5'-3-O-(thio)triphosphate binding assay. Activation of both transducin and Go was observed. The rate of Go activation was 18-fold greater than the rate of transducin activation. This indicates that the coupling of mGluR6 to Go is more efficient and suggests that Go may be involved in coupling to mGluR6 in ON-bipolar cells. PMID- 9407095 TI - Lysophosphatidic acid induces threonine phosphorylation of Tiam1 in Swiss 3T3 fibroblasts via activation of protein kinase C. AB - The Rho family of GTPases plays an important role in the control of cell shape, adhesion, movement, and growth. Several guanine nucleotide exchange factors have been identified that activate Rho family GTPases by promoting the binding of GTP to these proteins. However, little is known concerning the regulation of these GDP/GTP exchange factors. In this study, we demonstrate that lysophosphatidic acid (LPA) induces a rapid, sustainable phosphorylation of the Rac1-specific nucleotide exchange factor Tiam1 in Swiss 3T3 fibroblasts. LPA stimulated Tiam1 phosphorylation in a dose-dependent manner, and the protein was phosphorylated on threonine, but not tyrosine or serine. Tiam1 phosphorylation was also induced by platelet-derived growth factor, endothelin-1, bombesin, and bradykinin but not by epidermal growth factor. Significantly, pretreatment of Swiss 3T3 fibroblasts with 1 microM phorbol 12-myristate 13-acetate for 24 h, or with the selective protein kinase C inhibitor Ro-31-8220, reduced LPA-stimulated phosphorylation of Tiam1 by approximately 75%. Moreover, acute stimulation with 100 nM phorbol 12 myristate 13-acetate was sufficient to induce Tiam1 phosphorylation in vivo, and protein kinase C could phosphorylate purified Tiam1 on threonine residues in vitro. These data indicate that agonist-induced phosphorylation of Tiam1 is a general mechanism and suggest that it is likely to be important in its regulation. Protein kinase C appears to play a key role in phosphorylation of Tiam1. PMID- 9407096 TI - GFRalpha-2 and GFRalpha-3 are two new receptors for ligands of the GDNF family. AB - The receptor for glial cell line-derived neurotrophic factor (GDNF) consists of GFRalpha-1 and Ret. Neurturin is a GDNF-related neurotrophin whose receptor is presently unknown. Here we report that neurturin can bind to either GFRalpha-1 or GFRalpha-2, a novel receptor related to GFRalpha-1. Both GFRalpha-1 and GFRalpha 2 mediate neurturin-induced Ret phosphorylation. GDNF can also bind to either GFRalpha-1 or GFRalpha-2, and activate Ret in the presence of either binding receptor. Although both ligands interact with both receptors, cells expressing GFRalpha-1 bind GDNF more efficiently than neurturin, while cells expressing GFRalpha-2 bind neurturin preferentially. Cross-linking and Ret activation data also suggest that while there is cross-talk, GFRalpha-1 is the primary receptor for GDNF and GFRalpha-2 exhibits a preference for neurturin. We have also cloned a cDNA that apparently codes for a third member of the GFRalpha receptor family. This putative receptor, designated GFRalpha-3, is closely related in amino acid sequence and is nearly identical in the spacing of its cysteine residues to both GFRalpha-1 and GFRalpha-2. Analysis of the tissue distribution of GFRalpha-1, GFRalpha-2, GFRalpha-3, and Ret by Northern blot reveals overlapping but distinct patterns of expression. Consistent with a role in GDNF function, the GFRalphas and Ret are expressed in many of the same tissues, suggesting that GFRalphas mediate the action of GDNF family ligands in vivo. PMID- 9407097 TI - Alzheimer-like changes in microtubule-associated protein Tau induced by sulfated glycosaminoglycans. Inhibition of microtubule binding, stimulation of phosphorylation, and filament assembly depend on the degree of sulfation. AB - Hyperphosphorylated microtubule-associated protein tau is the major proteinaceous component of the paired helical and straight filaments which constitute a defining neuropathological characteristic of Alzheimer's disease and a number of other neurodegenerative disorders. We have recently shown that full-length recombinant tau assembles into Alzheimer-like filaments upon incubation with heparin. Heparin also promotes phosphorylation of tau by a number of protein kinases, prevents tau from binding to taxol-stabilized microtubules, and produces rapid disassembly of microtubules assembled from tau and tubulin. Here, we have used the above parameters to study the interactions between tau protein and a number of naturally occurring and synthetic glycosaminoglycans. We show that the magnitude of the glycosaminoglycan effects is proportional to their degree of sulfation. Thus, the strongly sulfated glycosaminoglycans dextran sulfate, pentosan polysulfate, and heparin were the most potent, whereas the non-sulfated dextran and hyaluronic acid were without effect. The moderately sulfated glycosaminoglycans heparan sulfate, chondroitin sulfate, and dermatan sulfate had intermediate effects, whereas keratan sulfate had little or no effect. These in vitro interactions between tau protein and sulfated glycosaminoglycans reproduced the known characteristics of paired helical filament-tau from Alzheimer's disease brain. Sulfated glycosaminoglycans are present in nerve cells in Alzheimer's disease brain in the early stages of neurofibrillary degeneration, suggesting that their interactions with tau may constitute a central event in the development of the neuronal pathology of Alzheimer's disease. PMID- 9407098 TI - cDNA cloning and characterization of a cis-retinol/3alpha-hydroxysterol short chain dehydrogenase. AB - We report a mouse cDNA that encodes a 317-amino acid short-chain dehydrogenase which recognizes as substrates 9-cis-retinol, 11-cis-retinol, 5alpha-androstan 3alpha,17beta-diol, and 5alpha-androstan-3alpha-ol-17-one. This cis retinol/androgen dehydrogenase (CRAD) shares closest amino acid similarity with mouse retinol dehydrogenase isozymes types 1 and 2 (86 and 91%, respectively). Recombinant CRAD uses NAD+ as its preferred cofactor and exhibits cooperative kinetics for cis-retinoids, but Michaelis-Menten kinetics for 3alpha hydroxysterols. Unlike recombinant retinol dehydrogenase isozymes, recombinant CRAD was inhibited by 4-methylpyrazole, was not stimulated by ethanol, and did not require phosphatidylcholine for optimal activity. CRAD mRNA was expressed intensely in kidney and liver, in contrast to retinol dehydrogenase isozymes, which show strong mRNA expression only in liver. CRAD mRNA expression was widespread (relative abundance): kidney (100) > liver (92) > small intestine (9) = heart (9) > retinal pigment epithelium and sclera (4.5) > brain (2) > retina and vitreous (1.6) > spleen (0.7) > testis (0.6) > lung (0.4). CRAD may catalyze the first step in an enzymatic pathway from 9-cis-retinol to generate the retinoid X receptor ligand 9-cis-retinoic acid and/or may regenerate dihydrotestosterone from its catabolite 5alpha-androstan-3alpha,17beta-diol. These data also illustrate the multifunctional nature of short-chain dehydrogenases and provide a potential mechanism for androgen-retinoid interactions. PMID- 9407099 TI - Diverse effects of BCL3 phosphorylation on its modulation of NF-kappaB p52 homodimer binding to DNA. AB - IkappaB proteins control the subcellular localization and DNA binding activity of NF-kappaB transcription factors. BCL3 is a nuclear IkappaB that can inhibit or enhance the binding of NF-kappaB p50 or p52 homodimers to consensus DNA-binding (kappaB) sequences or form a kappaB-binding complex with homodimers. To study BCL3 function, we have used gel shift analysis and tagged protein and tagged DNA coprecipitation analyses. Our results show that at intermediate ratios of BCL3 to p52 all observed phosphoforms of BCL3 are able to form a kappaB-binding complex with p52 homodimers. At low BCL3/p52 ratios, BCL3 increases the rate of p52 homodimer binding to kappaB sites in the presence of nonconsensus DNA and dissociates from the complex. At high BCL3/p52 ratios, BCL3 forms a higher order inhibitory complex with p52 homodimers. All of these effects depend on BCL3 phosphorylation and relative concentration. These results indicate that BCL3 phosphorylation may affect its regulation of NF-kappaB-dependent transcription in vivo. PMID- 9407100 TI - The Cbl phosphotyrosine-binding domain selects a D(N/D)XpY motif and binds to the Tyr292 negative regulatory phosphorylation site of ZAP-70. AB - The Cbl protooncogene product has emerged as a novel negative regulator of receptor and non-receptor tyrosine kinases through currently undefined mechanisms. Therefore, determining how Cbl physically interacts with tyrosine kinases is of substantial interest. We recently identified a phosphotyrosine binding (PTB) domain residing within the N-terminal transforming region of Cbl (Cbl-N), which mediated direct binding to ZAP-70 tyrosine kinase. Here, we have screened a degenerate phosphopeptide library and show that the Cbl-PTB domain selects a D(N/D)XpY motif, reminiscent of but distinct from the NPXpY motif recognized by the PTB domains of Shc and IRS-1/2. A phosphopeptide predicted by this motif and corresponding to the in vivo negative regulatory phosphorylation site of ZAP-70 (Tyr(P)292) specifically inhibited binding of ZAP-70 to Cbl-N. A ZAP-70/Y292F mutant failed to bind to Cbl-N, whereas a D290A mutant resulted in a 64% decrease in binding, confirming the importance of the Tyr(P) and Y-2 residues in Cbl-PTB domain recognition. Finally the ZAP-70/Y292F mutant also failed to associate with Cbl-N or full-length Cbl in vivo. These results identify a potential Cbl-PTB domain-dependent role for Cbl in the negative regulation of ZAP 70 and predict potential Cbl-PTB domain binding sites on other protein tyrosine kinases known to interact with Cbl. PMID- 9407101 TI - Switching of DNA secondary structure in proenkephalin transcriptional regulation. AB - Proper transcriptional regulation of the proenkephalin gene requires a switch between distinct factor binding sites that cannot exist at the same time. Each of the sites is formed from a nearly palindromic region that contains two functionally defined cAMP response elements. The region can switch between cruciform and linear duplex. Formation of the cruciform creates an alternative binding site for mediators of second messenger-directed transcription and abolishes the site present in the native duplex form. Use of the cruciform site has been shown to correlate with activated transcription. Analysis of DNA structure, protein binding, and gene expression from plasmids with mutant enhancers shows, however, that both sites are required for regulation of transcription. The two distinct structures form within the same enhancer. Shifting the balance between the two alters transcriptional response. PMID- 9407102 TI - Purification and determination of intact molecular mass by electrospray ionization mass spectrometry of the photosystem II reaction center subunits. AB - A reverse phase high pressure liquid chromatography purification system for the rapid separation of photosystem II reaction center proteins free of salts and detergents is described. This procedure results in the isolation of the three small subunits: alpha- and beta-subunits of cytochrome b559 and PsbI protein, with near base-line resolution between each peak, although the D1 and D2 proteins were partially deconvoluted. The molecular masses obtained by electrospray ionization mass spectrometry for the purified beta-subunit of cytochrome b559, alpha-subunit of cytochrome b559, and the PsbI protein, 4,394.8 +/- 0.4, 9,283.7 +/- 0.8, and 4,209.5 +/- 0.4 Da, respectively, are in excellent agreement with values obtained from previous characterization studies (Sharma, J., Panico, M., Barber, J., and Morris, H. R. (1997) J. Biol. Chem. 272, 3935-3943). Direct electrospray analysis of the D1 and D2 proteins suggests that these components exist in heterogeneous forms. The molecular mass ascribed to a predominant form of the D1 protein, 38, 040.9 +/- 6.5 Da, and the D2 protein, 39,456.1 +/- 7.7, are also in agreement with those expected for the mature nonphosphorylated states of these subunits. PMID- 9407103 TI - Primary structure characterization of the photosystem II D1 and D2 subunits. AB - Mass spectrometry techniques have been applied in a protein mapping strategy to elucidate the majority of the primary structures of the D1 and D2 proteins present in the photosystem II reaction center. Evidence verifying the post translational processing of the initiating methionine residue and acetylation of the free amino group, similar to those reported for other higher plant species, are presented for the two subunits from pea plants (Pisum sativum L.). Further covalent modifications observed on the D1 protein include the COOH-terminal processing with a loss of nine amino acids and phosphorylation of Thr2. In addition, the studies reported in this paper provide the first definitive characterization of oxidations on specific amino acids of the D1 and D2 proteins. We believe that these oxidations, and to a much lesser extent the phosphorylations, are major contributors to the heterogeneity observed during the electrospray analysis of the intact subunits reported in the accompanying paper (Sharma, J., Panico, M., Barber, J., and Morris, H. R. (1997) J. Biol. Chem. 272, 33153-33157). Significantly, all of the regions that have been identified as those particularly susceptible to oxidation are anticipated (from current models) to be in close proximity to the redox active components of the photosystem II complex. PMID- 9407104 TI - Genetic and biochemical evidence for a novel avermectin-sensitive chloride channel in Caenorhabditis elegans. Isolation and characterization. AB - Avermectins are a class of macrocyclic lactones that is widely used in crop protection and to treat helminth infections in man and animals. Two complementary DNAs (GluClalpha and GluClbeta) encoding chloride channels that are gated by avermectin and glutamate, respectively, were isolated from Caenorhabditis elegans. To study the role of these subunits in conferring avermectin sensitivity we isolated a mutant C. elegans strain with a Tc1 transposable element insertion that functionally inactivated the GluClalpha gene (GluClalpha::Tc1). GluClalpha::Tc1 animals exhibit a normal phenotype including typical avermectin sensitivity. Xenopus oocytes expressing GluClalpha::Tc1 strain mRNA elicited reduced amplitude avermectin and glutamate-dependent chloride currents. Avermectin binding assays in GluClalpha::Tc1 strain membranes showed the presence of high affinity binding sites, with a reduced Bmax. These experiments suggest that GluClalpha is a target for avermectin and that additional glutamate-gated and avermectin-sensitive chloride channel subunits exist in C. elegans. We isolated a cDNA (GluClalpha2) encoding a chloride channel that shares 75% amino acid identity with GluClalpha. This subunit forms homomeric channels that are gated irreversibly by avermectin and reversibly by glutamate. GluClalpha2 coassembles with GluClbeta to form heteromeric channels that are gated by both ligands. The presence of subunits related to GluClalpha may explain the low level and rarity of target site involvement in resistance to the avermectin class of compounds. PMID- 9407105 TI - Post-translational processing of RhoA. Carboxyl methylation of the carboxyl terminal prenylcysteine increases the half-life of Rhoa. AB - RhoA and related GTP-binding proteins are modified post-translationally at their carboxyl terminus to form a prenylcysteine methyl ester. The synthesis and post translational modification of RhoA and Cdc42 were examined in the RAW264 macrophage cell line, and the effect of carboxyl methylation on protein turnover was determined. Cells were labeled with [35S]cysteine, and RhoA or Cdc42 was immunoprecipitated with specific antibodies. Both RhoA and Cdc42 were methylated rapidly in control cells, with little accumulation of unmethylated protein. Carboxyl methylation of RhoA was inhibited by incubation of cells with a carbocyclic adenosine analog, 3-deazaaristeromycin, resulting in the accumulation of unmethylated RhoA. Under these conditions, Cdc42 methylation was inhibited only partially. When methylation was inhibited, the RhoA half-life decreased from 31 to 12 h, and the Cdc42 half-life decreased from 15 to 11 h. The increased degradation of unmethylated RhoA demonstrates a novel function for carboxyl terminal prenylcysteine carboxyl methylation in protecting RhoA and related proteins from degradation. PMID- 9407106 TI - Transforming growth factor-alpha enhances cyclin D1 transcription through the binding of early growth response protein to a cis-regulatory element in the cyclin D1 promoter. AB - Cyclin D1 is a critical oncogene involved in the regulation of progression through the G1 phase of the cell cycle, thereby contributing to cell proliferation. This is mediated through interaction of cyclin D1 with its catalytic partners, the cyclin-dependent kinases, and the subsequent phosphorylation of the retinoblastoma protein. Cyclin D1, in turn, is regulated by mitogenic stimuli. We demonstrate that transforming growth factor-alpha (TGFalpha) induces cyclin D1 mRNA in esophageal squamous epithelial cells, and this appears to correlate with increased cyclin D1 protein expression and cyclin dependent kinase 6 activity. The induction of cyclin D1 transcription by TGFalpha is mediated in part through the induction of the early growth response protein (Egr-1) and its subsequent binding of Egr-1 to a cis-regulatory region spanning nucleotides -144 to -104 of the cyclin D1 promoter. The Egr-1 binding activity to the cyclin D1 promoter appears to require de novo protein synthesis and is not influenced by Sp1 binding to overlapping Sp1 motifs. Taken together, these data provide evidence that TGFalpha enhances cyclin D1 transcription through the induction of Egr-1 binding to a cis-regulatory region in the cyclin D1 promoter. This has important mechanistic implications into the transcriptional regulation of cyclin D1 by an essential proproliferative growth factor and cell cycle progression. PMID- 9407107 TI - Purification, characterization, and localization of yeast Cox17p, a mitochondrial copper shuttle. AB - Cox17p was previously shown to be essential for the expression of cytochrome oxidase in Saccharomyces cerevisiae. In the present study COX17 has been placed under the control of the GAL10 promoter in an autonomously replicating plasmid. A yeast transformant harboring the high copy construct was used to purify Cox17p to homogeneity. Purified Cox17p contains 0.2-0.3 mol of copper per mol of protein. The molar copper content is increased to 1.8 after incubation of Cox17p in the presence of a 6-fold molar excess of cuprous chloride under reduced conditions. An antibody against Cox17p was obtained by immunization of rabbits with a carboxyl-terminal peptide coupled to bovine serum albumin. The antiserum detects Cox17p in both the mitochondrial and soluble protein fractions of wild type yeast and of the transformant overexpressing Cox17p. Exposure of intact mitochondria to hypotonic conditions causes most of Cox17p to be released as a soluble protein indicating that the mitochondrial fraction of Cox17p is localized in the intermembrane space. These results are consistent with the previously proposed function of Cox17p, namely in providing cytoplasmic copper for mitochondrial utilization. PMID- 9407108 TI - Apoptosis generates stable fragments of human type I keratins. AB - Type I and II keratins help maintain the structural integrity of epithelial cells. Since apoptosis involves progressive cell breakdown, we examined its effect on human keratin polypeptides 8, 18, and 19 (K8, K18, K19) that are expressed in simple-type epithelia as noncovalent type I (K18, K19) and type II (K8) heteropolymers. Apoptosis induces rapid hyperphosphorylation of most known K8/18 phosphorylation sites and delayed formation of K18 and K19 stable fragments. In contrast, K8 is resistant to proteolysis and remains associated with the K18 fragments. Transfection of phosphorylation/glycosylation-mutant K8 and K18 does not alter fragment formation. The protein domains of the keratin fragments were determined using epitope-defined antibodies, and microsequencing indicated that K18 cleavage occurs at a conserved caspase-specific aspartic acid. The fragments are found preferentially within the detergent-insoluble pool and can be generated, in a phosphorylation-independent manner, by incubating keratins with caspase-3 or with detergent lysates of apoptotic cells but not with lysates of nonapoptotic cells. Our results indicate that type I keratins are targets of apoptosis-activated caspases, which is likely a general feature of keratins in most if not all epithelial cells undergoing apoptosis. Keratin hyperphosphorylation occurs early but does not render the keratins better substrates of the downstream caspases. PMID- 9407109 TI - The complete cDNA sequence and structural polymorphism of the polypeptide chain of porcine submaxillary mucin. AB - The complete structure of the DNA encoding the polypeptide chain of porcine submaxillary mucin has been determined. The polypeptide is composed of distinct domains. A large central domain containing tandem repeats of 81 residues each is flanked by much shorter domains with sequences similar to the tandem repeats. Four disulfide-rich domains, three at the amino terminus and one at the carboxyl terminus, complete the chain. The disulfide-rich domains have significant sequence identity to those of other mucins and prepro-von Willebrand factor. The coding region of the mucin gene is highly polymorphic, and three alleles were identified in a single animal that encoded different numbers of the 81-residue tandem repeats. A single large exon devoid of introns encodes the tandem repeat domains. The largest allele with 135 tandem repeats encoded 13,288 amino acids to give a polypeptide with Mr = 1,184,106. The other two alleles contained 99 and 125 tandem repeats, respectively. Each allele also showed different restriction fragment length polymorphisms, which is consistent with the different patterns seen in individual animals. Fragment length polymorphism was also seen within two different families of animals, indicating that the polymorphism observed occurs in a single generation. PMID- 9407110 TI - Lack of correlation between in vitro and in vivo replication of precisely defined benz-a-anthracene adducted DNAs. AB - Like other polycyclic aromatic hydrocarbons, certain metabolites of benz[a]anthracene have been implicated as potent carcinogens. These effects are thought to be caused by the covalent binding of these species to nucleophilic groups on the bases of DNA. To address the molecular mechanisms by which these molecules induce mutations, this study employed oligonucleotides containing four site-specific N6 adenine-benz[a]anthracene diol epoxide adducts. Using a prokaryotic in vivo replication system, we have shown that both non-bay region anti-trans-benz[a]anthracene adducts are essentially nonmutagenic. In contrast, the bay region anti-trans-benz[a]anthracene lesions do induce point mutations at the adduct site. The mutagenic frequency of these bay region lesions is dependent on the stereochemistry about the adduct-forming bond, as well as the strain of Escherichia coli in which they are replicated. The ability of the bacterial replication machinery to bypass the lesions does not correlate with the differences observed in their mutagenesis. While both non-bay region adducts are readily bypassed in vivo, the bay region adducts are both blocking to approximately the same degree. In vitro studies of the interactions of E. coli DNA polymerase III with these adducts have also been undertaken to further dissect the relationship between adduct structure and biological activity. PMID- 9407111 TI - Enzymatic activity of 2'-5'-oligoadenylate synthetase is impaired by specific mutations that affect oligomerization of the protein. AB - Previous studies from our laboratory have shown that deletion of residues 321 to 344 of the 9-2 isozyme of 2'-5'-oligoadenylate (2-5(A)) synthetase causes a loss of its enzyme activity (Ghosh, S. K., Kusari, J., Bandyopadhyay, S. K., Samanta, H., Kumar, R., and Sen, G. C. (1991) J. Biol. Chem. 266, 15293-15299). Sequence comparison of this region among the different isozymes of 2-5(A) synthetases revealed that the residues at positions 330 to 333 are highly conserved. Alanine scanning mutagenesis of these residues demonstrated that the residues present at 331, 332, and 333 are important for activity but the proline at position 330 was dispensable. The triple mutant containing Ala residues at 331, 332, and 333 was completely inactive. Different double mutants were slightly active, and the three single mutants were partially active. The triple mutant was further characterized for delineating the nature of its defect. The mutant protein was enzymatically inactive irrespective of whether it was synthesized in rabbit reticulocyte lysate, Escherichia coli or Trichoplusia ni insect cells. It could bind double stranded RNA and ATP as efficiently as the wild type protein. It was, however, defective in oligomerization. Gel filtration and sedimentation velocity analyses of in vitro synthesized proteins revealed that the wild type protein, but not the triple mutant, formed tetramers. The tetrameric fraction, but not the monomeric fraction of the wild type protein was enzymatically active. The failure of the triple mutant to participate in homomeric protein-protein interaction was confirmed by in vivo assays in insect cells. These results indicate that tetramerization of the protein is required for the enzymatic activity of the small 2-5(A) synthetases. PMID- 9407112 TI - Cooperative and competitive protein interactions at the hsp70 promoter. AB - Drosophila heat shock factor (HSF) binds to specific sequence elements of heat shock genes and can activate their transcription 200-fold. Though HSF has an acidic activation domain, the mechanistic details of heat shock gene activation remain undefined. Here we report that HSF interacts directly with the general transcription factor TBP (TATA-box binding protein), and these two factors bind cooperatively to heat shock promoters. A third factor that binds heat shock promoters, GAGA factor, also interacts with HSF and further stabilizes HSF binding to heat shock elements (HSEs). The interaction of HSF and TBP is explored in some detail here and is shown to be mediated by residues in both the amino- and carboxyl-terminal portions of HSF. This HSF/TBP interaction can be specifically disrupted by competition with the potent acidic transcriptional activator VP16. We further show that the acidic domain of the largest subunit of Drosophila RNA polymerase II (Pol II) associates with TBP in vitro and is specifically displaced from TBP upon addition of HSF. The region of TBP that mediates both HSF and Pol II acidic domain binding maps to the conserved carboxyl terminal repeats and depends on at least one of the TBP residues known to be contacted by VP16 and to be critical for transcription activation. We discuss these findings in the context of a model in which HSF triggers hsp70 transcription by freeing the hsp70 promoter-paused Pol II from the constraints on elongation caused by the affinity of Pol II for general transcription factors. PMID- 9407113 TI - Dissection of C1q capability of interacting with IgG. Time-dependent formation of a tight and only partly reversible association. AB - C1q-bearing immune complexes have been observed in diseases such as rheumatoid arthritis and human immunodeficiency virus infection-associated neuropathy. For the purpose of understanding better the phenomenon of C1q-bearing immune complexes, we investigated the constancy of the C1q-IgG interaction. An enzyme linked immunosorbent assay was developed in which wells were coated with IgG to mimic antigen-complexed IgG. Serial dilutions of C1q were applied for distinct time intervals, and bound C1q was detected either directly or after exposure to one of several elution buffers. Our results show that a part of C1q attached to IgG forms a tight association that is not reversible under treatment with buffers containing usually protein-protein interaction-dissociating reagents such as 3 M NaCl, 5 M urea, sodium dodecyl sulfate, or beta-mercaptoethanol. The formation of the highly stable C1q-IgG complex was found to be time-, temperature-, and pH dependent and to proceed with bound C1q even in the absence of free C1q in the supernatant. In ligand blotting experiments we demonstrate for the first time directly that all three chains of C1q can individually bind IgG. Altogether, our results provide a suitable explanation for the formation and persistence of C1q bearing immune complexes. PMID- 9407114 TI - Reversible disruption of cell-matrix and cell-cell interactions by overexpression of sialomucin complex. AB - Sialomucin complex (SMC) is a large, heterodimeric glycoprotein complex composed of mucin (ASGP-1) and transmembrane (ASGP-2) subunits and expressed abundantly on the cell surface of ascites 13762 rat mammary adenocarcinoma cells. We have isolated recombinant cDNAs containing different numbers of ASGP-1 mucin repeats, which can be expressed as protein products with variable lengths. To study the anti-adhesive effect of SMC, these cDNAs were transfected into human cancer cell lines. Using a tetracycline-responsive, inducible expression system, we demonstrated that the overexpression of SMC induces morphology changes, cell detachment, and cell-cell dissociation of transfected A375 human melanoma cells in culture. The transition between the adherent and suspension states of the cells is fully reversible and dependent on the SMC expression level. The anti adhesion effect of SMC was further analyzed kinetically by measuring the cell adhesion of transfected A375 melanoma and MCF-7 breast cancer cell lines to fibronectin, laminin, and collagen IV, demonstrating that SMC disrupts integrin mediated cell adhesion to extracellular matrix proteins. The degree of this anti adhesion effect was dependent on the number of mucin repeats in the SMC molecule as well as the level of cell surface expression. PMID- 9407115 TI - Functional overlap of tRNA nucleotidyltransferase, poly(A) polymerase I, and polynucleotide phosphorylase. AB - Repair of the 3'-terminal -CCA sequence of tRNA generally requires the action of the enzyme tRNA nucleotidyltransferase. However, in Escherichia coli in the absence of this enzyme, a decreased level of tRNA end repair continues. To ascertain the enzymes responsible for this residual repair, mutant strains were constructed lacking tRNA nucleotidyltransferase and other enzymes potentially involved in the process, poly(A) polymerase I and polynucleotide phosphorylase (PNPase). Strains lacking tRNA nucleotidyltransferase and either one of the other enzymes displayed decreased growth rates and increased levels of defective tRNA compared with the single cca mutant. Triple mutants lacking all three enzymes grew very slowly, had even more defective tRNA, and were devoid of activity incorporating AMP into tRNA-C-C. Overexpression of poly(A) polymerase I, but not PNPase, partially compensated for the absence of tRNA nucleotidyltransferase. These data show that poly(A) polymerase I and PNPase participate in the end repair process and are required to maintain functional tRNA levels when tRNA nucleotidyltransferase is absent. PMID- 9407116 TI - The Src family kinase Hck interacts with Bcr-Abl by a kinase-independent mechanism and phosphorylates the Grb2-binding site of Bcr. AB - bcr-abl, the oncogene causing chronic myeloid leukemia, encodes a fusion protein with constitutively active tyrosine kinase and transforming capacity in hematopoietic cells. Various intracellular signaling intermediates become activated and/or associate by/with Bcr-Abl, including the Src family kinase Hck. To elucidate some of the structural requirements and functional consequences of the association of Bcr-Abl with Hck, their interaction was investigated in transiently transfected COS7 cells. Neither the complex formation of Hck kinase with Bcr-Abl nor the activation of Hck by Bcr-Abl was dependent on the Abl kinase activity. Both inactivating point mutations of Hck and dephosphorylation of Hck enhanced its complex formation with Bcr-Abl, indicating that their physical interaction was negatively regulated by Hck (auto)phosphorylation. Finally, experiments with a series of kinase negative Bcr-Abl mutants showed that Hck phosphorylated Bcr-Abl and induced the binding of Grb2 to Tyr177 of Bcr-Abl. Taken together, our results suggest that Bcr-Abl preferentially binds inactive forms of Hck by an Abl kinase-independent mechanism. This physical interaction stimulates the Hck tyrosine kinase, which may then phosphorylate the Grb2-binding site in Bcr-Abl. PMID- 9407117 TI - A dominant-negative form of transcription factor MEF2 inhibits myogenesis. AB - A biological role for MEF2 (myocyte enhancer factor 2) activity during mammalian myogenesis has been inferred but not directly proven because of its role in the transcriptional activation of many muscle-specific genes. Therefore, our purpose was to determine whether MEF2 activity is absolutely required for mammalian myogenesis. Using a dominant-negative approach to address this question, we constructed a mutated MEF2A protein comprised of the amino-terminal DNA binding/dimerization domain of MEF2A without its trans-activation domain as a bacterial fusion protein (GST-131) or in a eukaryotic expression vector (pcDNA 131). GST-131 and the protein encoded by pcDNA-131 bind specifically to the MEF2 cis element and abrogate trans-activation of a MEF2-responsive luciferase reporter gene by wild type MEF2A, thus serving a role as trans-dominant inhibitors of MEF2 function. In congruence with their ability to interfere with wild type MEF2 function, microinjection of GST-131 or pcDNA-131 into L6E9 or C2C12 myoblasts inhibited myotube formation. Immunofluorescence analysis showed that the expression of myogenin, myosin heavy chain, and MEF2A were inhibited in the GST-131 or pcDNA-131-injected cells compared with GST or pcDNA-injected controls. We also document that this trans-dominant MEF2 inhibitor impairs the myogenic conversion of C3H10T1/2 fibroblasts by MyoD. Thus, these data provide evidence that the trans-activation function of the MEF2 proteins during mammalian myogenesis is required for muscle-specific gene expression and differentiation. PMID- 9407118 TI - Expression, purification, and metal binding properties of the N-terminal domain from the wilson disease putative copper-transporting ATPase (ATP7B). AB - The putative copper binding domain from the copper-transporting ATPase implicated in Wilson disease (ATP7B) has been expressed and purified as a fusion to glutathione S-transferase. Immobilized metal ion affinity chromatography revealed that the fusion protein is able to bind to columns charged with different transition metals with varying affinities as follows: Cu(II)>>Zn(II)>Ni(II)>Co(II). The fusion protein did not bind to columns charged with Fe(II) or Fe(III). 65Zinc(II) blotting analysis showed that the domain is able to bind Zn(II) over a range of pH values from 6.5 to 9.0. Competition 65Zn(II) blotting showed that Cd(II), Hg(II), Au(III), and Fe(III) can successfully compete with Zn(II), at comparable concentrations, for binding to the domain. In contrast, the domain had little or no affinity for Ca(II), Mg(II), Mn(II), and Ni(II) relative to copper. Neutron activation analysis of the copper bound to the domain showed a copper:protein ratio of 6.5-7.3:1. Both Cu(II) and Cu(I) were found to have a higher affinity for the domain relative to Zn(II). In addition, a sharp, reproducible transition was only observed in competition experiments with copper, which may suggest that copper binding has some degree of cooperativity. PMID- 9407119 TI - Hsp70 and Hsp40 chaperone activities in the cytoplasm and the nucleus of mammalian cells. AB - The existence and function of a Hsp40-Hsp70 chaperone machinery in mammalian cells in vivo was investigated. The rate of heat inactivation of firefly luciferase transiently expressed in hamster O23 fibroblasts was analyzed in cells co-transfected with the gene encoding the human Hsp40 (Ohtsuka, K. (1993) Biochem. Biophys. Res. Commun. 197, 235-240), the human inducible Hsp70 (Hunt, C., and Morimoto, R. I. (1985) Proc. Natl. Acad. Sci. U. S. A. 82, 6455-6459), or a combination of both. Whereas the expression of human Hsp70 alone in hamster cells was sufficient for the protection of firefly luciferase during heat shock, expression of the human Hsp40 alone was not. Rather, this led to a small but significant increase in the heat sensitivity of luciferase. The expression of the human Hsp40 only led to heat protection when the human Hsp70 was expressed as well. Under such conditions the rate of luciferase reactivation from the heat inactivated state was increased, but the rate of inactivation during heat shock was not affected. Using constructs that direct firefly luciferase either to the cytoplasm or to the nucleus (Michels, A. A., Nguyen, V.-T., Konings, A. W. T., Kampinga, H. H., and Bensaude, O. (1995) Eur. J. Biochem. 234, 382-389), it was demonstrated that these chaperone functions are found in both compartments. Our data provide the first evidence on how the Hsp40/Hsp70 chaperone complex acts as heat protector in mammalian cells in vivo. PMID- 9407120 TI - Recombinant subunits of mammalian elongation factor 1 expressed in Escherichia coli. Subunit interactions, elongation activity, and phosphorylation by protein kinase CKII. AB - The first step in elongation requires two different activities; elongation factor (EF)-1alpha transfers aminoacyl-tRNA to the ribosome and is released upon hydrolysis of GTP, EF-1betagammadelta catalyzes exchange of GDP on EF-1alpha with GTP. To analyze the role of the individual subunits of EF-1 in elongation, the cDNAs for the beta, gamma, and delta subunits of EF-1 from rabbit were cloned, and proteins of 225, 437, and 280 amino acids, respectively, were expressed in Escherichia coli. The purified recombinant beta subunit migrates as a dimer and the gamma subunit as a trimer upon gel filtration, whereas the delta subunit forms a large aggregate. Complexes of betagamma, gammadelta and betagammadelta were formed by self-association and eluted with a molecular mass of approximately 160, 530, and 670 kDa, respectively; no interaction was observed between beta and delta. The activity of the recombinant subunits was determined with native EF 1alpha by measuring stimulation of the rate of elongation by poly(U)-directed polyphenylalanine synthesis. Recombinant beta and delta alone stimulated the rate of elongation by 10-fold, with a ratio of 5alpha:2beta or delta. The betagammadelta complex stimulated EF-1alpha activity up to 10-fold with a ratio of 20alpha to 1betagammadelta. Phosphorylation of the beta and delta subunits alone or in betagammadelta by protein kinase CKII had no effect on the rate of elongation. PMID- 9407121 TI - Gene characterization, promoter analysis, and chromosomal localization of human bleomycin hydrolase. AB - The human gene encoding bleomycin hydrolase, a cysteine proteinase involved in chemotherapy resistance, has been cloned, and its overall organization has been established. The gene is composed of 12 coding exons and 11 introns and spans more than 30 kilobases. The number and distribution of exons and introns differ from those reported for other human cysteine proteinases, indicating that these genes are only distantly related. Nucleotide sequence analysis of about 1.2 kilobases of the 5'-flanking region of the human bleomycin hydrolase gene revealed a high GC content, a ratio of CpG/GpC close to unity, and the absence of consensus transcriptional sequences such as TATA box or CCAAT box. All these features are characteristic of housekeeping genes and provide an explanation for the widespread expression of bleomycin hydrolase in human tissues. The 5' flanking region of the gene also contains a polymorphic CCG trinucleotide repeat that may be a target of genetic instability events and affect its transcriptional activity. Chromosomal localization of the human bleomycin hydrolase gene revealed that it maps to chromosome 17q11.2, very close to the locus of the neurofibromatosis type 1 gene. This location is unique for any cysteine proteinase mapped to date. Finally, Southern blot analysis of DNA from leukocytes and autologous breast tumors has shown that the bleomycin hydrolase gene is not a frequent target of amplification in human breast carcinomas. PMID- 9407122 TI - Determination of the transmembrane topology of herpes simplex virus type 1 glycoprotein K. AB - Herpes simplex virus type 1 glycoprotein K (gK) plays an essential role in viral replication and cell fusion. gK is a very hydrophobic membrane protein that contains a signal sequence and several hydrophobic regions. It has been shown that mutations inducing cell fusion map to two distinct domains of gK, suggesting that these domains are functionally important. To understand the transmembrane topology of gK and the localization of these functional domains, we constructed a set of gK deletion, insertion, and truncation mutants and expressed these by in vitro translation in the presence of microsomal membranes. The transmembrane topology of gK was determined by examination of the post-translational processing and protease sensitivity of the mutant proteins. Our data demonstrate that gK contains three transmembrane domains (amino acids 125-139, 226-239, and 311-325). Another hydrophobic domain (amino acids 241-265), which is relatively less hydrophobic and much longer compared with the transmembrane sequences, is located in the extracellular loop. The analysis showed that the domains containing syncytial mutations are both ectodomains. They may interact with each other to form a complex tertiary structure that is critical for the biological function of gK. PMID- 9407123 TI - Helicase delivery and activation by DnaA and TrfA proteins during the initiation of replication of the broad host range plasmid RK2. AB - Specific binding of the plasmid-encoded protein, TrfA, and the Escherichia coli DnaA protein to the origin region (oriV) is required for the initiation of replication of the broad host range plasmid RK2. It has been shown that the DnaA protein which binds to DnaA boxes upstream of the TrfA-binding sites (iterons) cannot by itself form an open complex, but it enhances the formation of the open complex by TrfA (Konieczny, I., Doran, K. S., Helinski, D. R., Blasina, A. (1997) J. Biol. Chem. 272, 20173). In this study an in vitro replication system is reconstituted from purified TrfA protein and E. coli proteins. With this system, a specific interaction between the DnaA and DnaB proteins is required for delivery of the helicase to the RK2 origin region. Although the DnaA protein directs the DnaB-DnaC complex to the plasmid replication origin, it cannot by itself activate the helicase. Both DnaA and TrfA proteins are required for DnaB induced template unwinding. We propose that specific changes in the nucleoprotein structure mediated by TrfA result in a repositioning of the DnaB helicase within the open origin region and an activation of the DnaB protein for template unwinding. PMID- 9407124 TI - Inhibition of the iron-induced ZmFer1 maize ferritin gene expression by antioxidants and serine/threonine phosphatase inhibitors. AB - Two pathways have been implicated in the regulation of maize ferritin synthesis in response to iron. One of them involves the plant hormone abscisic acid (ABA) and controls the expression of ZmFer2 gene(s). Another pathway, ABA-independent, has been characterized in a de-rooted maize plantlet system and involves an oxidative step. The ZmFer1 maize ferritin gene is not regulated by ABA, and it is shown in this paper that the corresponding mRNA accumulates in de-rooted maize plantlets and BMS (Black Mexican Sweet) maize cell suspension cultures in response to iron via the oxidative pathway described previously. To investigate ZmFer1 gene regulation further, the BMS cell system has been used to develop a transient expression assay using a ZmFer1-beta-glucuronidase fusion. Both iron induction and antioxidant inhibition of ZmFer1 gene expression were observed in this system. Using Northern blot analysis and transient expression experiments, it was shown that both okadaic acid and calyculin A, two serine/ threonine phosphatase inhibitors, specifically inhibit ZmFer1 gene expression. These data indicate that an okadaic acid-sensitive protein phosphatase activity is involved in the regulation of the ZmFer1 ferritin gene in maize cells, and this activity is required for iron-induced expression of this gene. PMID- 9407125 TI - Differential phosphorylation of T-47D human breast cancer cell substrates by D1-, D3-, E-, and A-type cyclin-CDK complexes. AB - The cyclin-dependent kinases (CDKs) promote cell cycle transitions in mammalian cells by phosphorylation of key substrates. To characterize substrates of the G1 and S phase cyclin-CDK complexes, including cyclin D1-CDK4, cyclin D3-CDK4, cyclin D3-CDK6, cyclin E-CDK2, and cyclin A-CDK2, which are largely undefined, we phosphorylated T-47D breast cancer cell nuclear lysates partially purified by ion exchange chromatography with purified baculovirus expressed cyclin-CDK complexes. A comparison of the substrates that were phosphorylated by the different cyclin D CDKs revealed some common as well as specific substrates. Hence, cyclin D1-CDK4 specifically phosphorylated a 38-kDa protein while cyclin D3-CDK4 specifically phosphorylated proteins of 105, 102, and 42 kDa. A 24-kDa protein was phosphorylated by both complexes. Cyclin D3-CDK6 exhibited similar substrate preferences to cyclin D3-CDK4, phosphorylating the 105- and 102-kDa proteins but not the 24-kDa protein. Hence, both the cyclin D1 and D3 as well as CDK4 and CDK6 subunits can confer substrate specificity on the overall cyclin D-CDK complex. Cyclin E-CDK2 and cyclin A-CDK2 phosphorylated a greater number of substrates than the cyclin D-CDKs, ranging in size from 10 kDa to over 200 kDa. Twenty-two substrates were common to both complexes, while six were specific for cyclin A CDK2 and only one protein of 34 kDa was specific for cyclin E-CDK2. These studies indicate that cyclins E and A modulate the specificity of CDK2 and have demonstrated substrates that may be important for the specific roles of these cyclin-CDKs during G1 and S phase progression. Protein sequencing of one of the cyclin-CDK substrates characterized in this study identified this protein as nucleolin, a previously characterized CDC2 (CDK1) substrate, thus indicating the utility of this approach in identifying cyclin-CDK targets. These results show that both the cyclin and CDK subunits can regulate the substrate specificity of the overall cyclin-CDK complex and have demonstrated numerous substrates of D-, E , and A-type cyclin-CDK complexes potentially involved in regulating transit through the G1 and S phases of the cell cycle. PMID- 9407126 TI - The catalytic domain of protein kinase Cdelta confers protection from down regulation induced by bryostatin 1. AB - Bryostatin 1 (Bryo) has been shown to induce biphasic dose-response curves for down-regulating protein kinase Cdelta (PKCdelta) as well as for protecting PKCdelta from down-regulation induced by phorbol 12-myristate 13-acetate (PMA). To identify regions within PKCdelta that confer these responses to Bryo, we utilized reciprocal PKCalpha and PKCdelta chimeras (PKCalpha/delta and PKCdelta/alpha) constructed by exchanging the regulatory and catalytic domains of these PKCs. These chimeras and wild-type PKCalpha/alpha and PKCdelta/delta constructed in the same way were stably expressed in NIH 3T3 fibroblasts. Twenty four h of treatment with Bryo induced a biphasic dose-response curve for down regulating both wild-type PKCdelta/delta and the PKCalpha/delta chimera. In contrast, Bryo led to a nearly complete down-regulation of both PKCalpha/alpha and PKCdelta/alpha and also produced a faster mobility form of these species on SDS-polyacrylamide gel electrophoresis. The nature of both the regulatory and, to a lesser extent, the catalytic domains affected the potency of Bryo to down regulate the chimeric PKC proteins as well as to protect PKCalpha/delta and PKCdelta/delta from down-regulation. Bryo at high concentrations also inhibited the down-regulation of PKCdelta/delta and PKCalpha/delta induced by 1 microM PMA when co-applied. The portion of PKC protected by Bryo from down-regulation by either Bryo or PMA was localized in the particulate fraction of the cells. We conclude that the catalytic domain of PKCdelta confers protection from down regulation induced by Bryo or Bryo plus PMA, suggesting that this domain contains the isotype-specific determinants involved in the unique effect of Bryo on PKCdelta. PMID- 9407127 TI - Characterization of two protein activities that interact at the promoter of the trypanosomatid spliced leader RNA. AB - All trypanosome mRNAs have a spliced leader (SL). The SL RNA gene in Leptomonas seymouri is a member of the small nuclear RNA gene family. However, the SL RNA is required in stoichiometric amounts for trans-splicing during mRNA formation. Expression of the SL RNA gene requires sequence elements at bp -60 to -70 and bp 30 to -40 upstream from the transcription initiation site. Using conventional and affinity chromatography, we have identified and characterized an approximately 122-kDa protein, promoter-binding protein (PBP) -1, that binds to double-strand DNA. The PBP-1-binding site is within the bp -60 to -70 element determined by DNase I footprinting. Therefore, PBP-1 is the first characterized double-strand DNA binding activity that interacts with a trypanosome gene promoter. A second protein, PBP-2, interacts with the PBP-1:DNA complex and its DNase I footprint extends to include the second promoter element (bp -30 to -40). An alteration of the spacing between the two promoter elements or mutation of the second element decreases PBP-2/PBP-1:DNA stability. Taken together, these data suggest that PBP 1 and PBP-2 are components of a transcription initiation complex that assembles within the SL RNA gene promoter. PMID- 9407128 TI - The zinc finger protein CTCF binds to the APBbeta domain of the amyloid beta protein precursor promoter. Evidence for a role in transcriptional activation. AB - The promoter of the amyloid beta-protein precursor (APP) gene directs high levels of cell type-specific transcription with 94 base pairs 5' to the main transcriptional start site. An essential activator domain in this proximal APP promoter is a nuclear factor binding site designated as APBbeta. The recognition domain for the APBbeta binding factor is located between position -93 and -82 relative to the main transcriptional start site. The nuclear factor that binds to the APBbeta site was partially purified by multiple steps of ion exchange and hydroxyapatite chromatography. Based on UV cross-linking results, a protein with an apparent molecular mass of 140 kDa was selected as the putative APBbeta binding protein. After the final purification step consisting of preparative SDS polyacrylamide gel electrophoresis, partial peptide sequences were obtained that completely matched the transcriptional factor CTCF. This protein is a known regulator of c-myc and lysozyme gene expression, and it binds to a variety of diverse DNA sequences. The binding of CTCF to the APBbeta domain was further established by competition with CTCF binding oligonucleotides in mobility shift electrophoresis. The identity was also confirmed by the observation that the APBbeta binding factor is recognized by antibodies against C- and N-terminal sequences of CTCF. In addition, oligonucleotide competition during in vitro transcription affirmed that CTCF acts as a transcriptional activator in the APP gene promoter. PMID- 9407129 TI - Estrogen up-regulates apolipoprotein E (ApoE) gene expression by increasing ApoE mRNA in the translating pool via the estrogen receptor alpha-mediated pathway. AB - The antiatherogenic property of estrogens is mediated via at least two mechanisms: first by affecting plasma lipoprotein profiles, and second by affecting the components of the vessel wall. Raising plasma apolipoprotein E (apoE) in mice protects them against diet-induced atherosclerosis (Shimano, H., Yamada, N., Katsuki, M., Gotoda, T., Harada, K., Murase, T., Fukuzawa, C., Takaku, F., and Yazaka, Y. (1992) Proc. Natl. Acad. Sci. U. S. A. 89, 1750-1754). It is possible that estrogen may be antiatherogenic at least in part by increasing plasma apoE levels. Therefore, we studied the regulation of apoE by estrogen. A survey of 15 inbred strains of mice showed that some mouse strains responded to injections or subcutaneously implanted pellets of estradiol by raising their apoB and apoE levels and some did not. We performed detailed studies in two "responder" strains, C57L and C57BL, and two "non-responder" strains, C3H and BALBc. Responders increased their plasma apoE levels 2.5-fold. Non-responders' levels were altered +/-10%. In the responders the distribution of apoE among the plasma lipoproteins shifted from high density lipoprotein toward the apoB-containing lipoprotein fractions. In nonresponders the shift was toward high density lipoprotein. Hepatic apoE mRNA levels and relative rates of apoE mRNA transcription were unchanged in all strains, suggesting that apoE regulation occurred at posttranscriptional loci. Therefore, we measured apoE synthesis in fresh liver slices and on isolated hepatic polysomes. Two-fold increases were noted but only in responders accompanied by selective 1.5-fold increases in polysomal apoE mRNA levels. Similar increases in apoE synthesis were also observed in castrated C57BL mice given either physiological or pharmacological replacement doses of estradiol, but not testosterone, suggesting that the effect of estradiol was specific on the distribution of apoE mRNA in the translationally active polysomal pool. Next, we examined whether the effects of estrogen on apoE translation were mediated by estrogen receptors (ER). ER-alpha knock-out mice and their wild-type littermates were administered estradiol. As expected, apoE levels and hepatic apoE synthesis increased more than 2-fold in the wild-type littermates, but only 20% increases in the plasma apoE and hepatic synthesis were observed in the ER knock-out mice. Hepatic apoE mRNA levels did not change in either the wild-type or the ER knock-out mice. Thus, estradiol up-regulates apoE gene expression by increasing levels of apoE mRNA in the polysomal translating pool. Furthermore, the increased polysomal recruitment of apoE mRNA is largely mediated by estrogen receptors. PMID- 9407130 TI - Transcriptional activation of the Glut1 gene in response to oxidative stress in L6 myotubes. AB - Exposure of L6 myotubes to prolonged low grade oxidative stress results in increased Glut1 expression at both the protein and mRNA levels, leading to elevated glucose transport activity. To further understand the cellular mechanisms responsible for this adaptive response, the Glut1 transcription rate and mRNA stability were assessed. Nuclear run-on assays revealed 2.0- and 2.4 fold increases in Glut1 transcription rates in glucose oxidase- and xanthine/xanthine oxidase-pretreated cells, respectively. Glut1 mRNA stability was increased with both treatments compared with the control (t1/2 = 7.8 +/- 1.3, 6.0 +/- 2.0, and 2.4 +/- 0.5 h, respectively). The serum-responsive element and AP-1 (but not the cAMP-responsive element) showed increased binding capacity following oxidative stress. Both activation of AP-1 binding and elevation of Glut1 mRNA were prevented by cycloheximide. The involvement of enhancer 1 of the Glut1 gene was demonstrated using transfected 293 cells. Induction of Glut1 mRNA in response to oxidative stress differed from its activation by chronic insulin exposure as demonstrated by the ability of rapamycin to inhibit the latter without an effect on the former. In conclusion, oxidative stress increases the Glut1 transcription rate by mechanisms that may involve activation of AP-1 binding to enhancer 1 of the Glut1 gene. PMID- 9407131 TI - The bovine papillomavirus E6 protein binds to the LD motif repeats of paxillin and blocks its interaction with vinculin and the focal adhesion kinase. AB - The bovine papillomavirus type 1 (BPV-1) E6 oncoprotein can transform fibroblasts and induce anchorage-independent growth and disassembly of the actin stress fibers. We have previously shown that the E6 protein interacts with the focal adhesion protein, paxillin, suggesting a direct role of E6 in the disruption of the actin cytoskeleton. We have now mapped the E6 binding sites on paxillin to the LD motif repeats region, which has been implicated in mediating paxillin binding to two other focal adhesion proteins, vinculin and the focal adhesion kinase. The five LD motif repeats identified in paxillin do not contribute equally to its interaction with E6. The first LD repeat is most critical for paxillin binding to E6 both in vitro and in vivo. Furthermore, the binding of recombinant wild-type E6 protein to paxillin blocked the interaction of several cellular proteins with paxillin, including vinculin and the focal adhesion kinase. A mutant E6 protein (H105) which does not bind to paxillin had no effect on the binding of these cellular proteins to paxillin. These data suggest that E6 disruption of the actin stress fibers occurs through blocking the interaction of paxillin with its cellular effectors such as vinculin and the focal adhesion kinase. PMID- 9407132 TI - Calcium-dependent signaling pathways in T cells. Potential role of calpain, protein tyrosine phosphatase 1b, and p130Cas in integrin-mediated signaling events. AB - Engagement of beta1 integrin receptors initiates an increase in intracellular calcium concentrations in T cells, potentially affecting calcium-sensitive signaling pathways. The calcium-activated cysteine protease, calpain, regulates a variety of cell functions by calcium-dependent limited proteolysis. To investigate the function of calpain in T cells, we sought to determine the role of this protease in calcium-dependent signaling events. Subsequent to elevations in intracellular calcium concentrations induced by ionomycin or adherence to fibronectin, calpain activity translocated to the cytoskeletal/membrane fraction of T cells. In addition, stimulation of T cells with these agents initiated the proteolytic cleavage of protein tyrosine phosphatase 1B by calpain. Enzymatic cleavage of protein tyrosine phosphatase 1B occurs near the endoplasmic reticulum targeting sequence and results in the generation of an enzymatically active form of the phosphatase. Furthermore, we show that both the native and the cleaved forms of protein tyrosine phosphatase 1B interact with p130(Cas) in T cells. This interaction may serve to relocate protein tyrosine phosphatase 1B to sites of focal contact resulting in potential interactions with substrates previously inaccessible to the endoplasmic reticulum-associated phosphatase. Thus, we describe a novel calcium-dependent signaling pathway in T cells that may mediate signals generated by beta1 integrin adherence to the extracellular matrix. PMID- 9407133 TI - Proteolysis and tyrosine phosphorylation of p34cdc2/cyclin B. The role of MCM2 and initiation of DNA replication to allow tyrosine phosphorylation of p34cdc2. AB - Previously, it has been shown that Aspergillus cells lacking the function of nimQ and the anaphase-promoting complex (APC) component bimEAPC1 enter mitosis without replicating DNA. Here nimQ is shown to encode an MCM2 homologue. Although mutation of nimQMCM2 inhibits initiation of DNA replication, a few cells do enter mitosis. Cells arrested at G1/S by lack of nimQMCM2 contain p34(cdc2)/cyclin B, but p34(cdc2) remains tyrosine dephosphorylated, even after DNA damage. However, arrest of DNA replication using hydroxyurea followed by inactivation of nimQMCM2 and bimEAPC1 does not abrogate the S phase arrest checkpoint over mitosis. nimQMCM2, likely via initiation of DNA replication, is therefore required to trigger tyrosine phosphorylation of p34(cdc2) during the G1 to S transition, which may occur by inactivation of nimTcdc25. Cells lacking both nimQMCM2 and bimEAPC1 are deficient in the S phase arrest checkpoint over mitosis because they lack both tyrosine phosphorylation of p34(cdc2) and the function of bimEAPC1. Initiation of DNA replication, which requires nimQMCM2, is apparently critical to switch mitotic regulation from the APC to include tyrosine phosphorylation of p34(cdc2) at G1/S. We also show that cells arrested at G1/S due to lack of nimQMCM2 continue to replicate spindle pole bodies in the absence of DNA replication and can undergo anaphase in the absence of APC function. PMID- 9407134 TI - Novel exonic elements that modulate splicing of the human fibronectin EDA exon. AB - Three exons in the fibronectin primary transcript are alternatively spliced in a tissue- and developmental stage-specific manner. One of these exons, EDA, has been shown previously by others to contain two splicing regulatory elements between 155 and 180 nucleotides downstream of the 3'-splice site: an exon splicing enhancer and a negative element. By transient expression of a chimeric beta-globin/fibronectin EDA intron in COS-7 cells, we have identified two additional exonic splicing regulatory elements. RNA generated by a construct containing the first 120 nucleotides of the fibronectin EDA exon was spliced with an efficiency of approximately 50%. Deletion of most of the fibronectin EDA exon sequences resulted in a 20-fold increase in the amount of spliced RNA, indicative of an exon splicing silencer. Deletion and mutagenesis studies suggest that the fibronectin exon splicing silencer is associated with a conserved RNA secondary structure. In addition, sequences between nucleotides 93 and 118 of the EDA exon contain a non-purine-rich splicing enhancer as demonstrated by its ability to function in a heterologous context. PMID- 9407135 TI - The role of CDP-diacylglycerol synthetase and phosphatidylinositol synthase activity levels in the regulation of cellular phosphatidylinositol content. AB - The regulation of phosphatidylinositol synthesis was examined by cloning and expressing in COS-7 cells the human cDNAs encoding the two enzymes in the biosynthetic pathway. Human CDP-diacylglycerol synthetase (cds1) and phosphatidylinositol synthase (pis1) clones were identified in the human expressed sequence-tagged (EST) data base, and full-length cDNAs were obtained by library screening. The cds1 cDNA did not possess a recognizable mitochondrial import signal, and the activity of the expressed Cds1 protein was stimulated by nucleoside triphosphates in vitro, indicating that cds1 did not encode the mitochondrial-specific isozyme. There were two mRNA species (3.9 and 5.6 kilobases) detected on Northern blots hybridized with the cds1 probe that were expressed at distinctly different levels in various human tissues. Consistent with the presence of the two mRNAs, a cDNA predicted to encode a second human CDP diacylglycerol synthetase (cds2) was also uncovered in the EST data base. In contrast to the two cds mRNAs, a single, 2.1-kilobase pis1 mRNA was uniformly expressed in all human tissues examined. Expression of the pis1 gene led to the overproduction of both phosphatidylinositol synthase and phosphatidylinositol:inositol exchange reactions, indicating that the Pis1 polypeptide catalyzed both of these activities. Phosphatase treatment of cell extracts abolished the CMP-independent phosphatidylinositol:inositol exchange reaction, and exchange activity was completely restored by the addition of CMP. Overexpression of cds1 or pis1 alone or in combination did not enhance the rate of phosphatidylinositol biosynthesis. Also, overexpression did not result in a significant proportional increase in the cellular levels of CDP-diacylglycerol or phosphatidylinositol. These data illustrate that the levels of Cds1 and Pis1 protein expression are not critical determinants of cellular PtdIns content and argue against a determining role for the activity of either of these enzymes in the regulation of PtdIns biosynthesis. PMID- 9407136 TI - Norrie disease protein (norrin) forms disulfide-linked oligomers associated with the extracellular matrix. AB - COS-7 cells transfected with a DNA construct encoding the 133 amino acids in norrin plus six histidine residues at its carboxyl terminus were pulse-labeled with [35S]cysteine, and the labeled norrin was examined in cell lysates, the medium, and the extracellular matrix. SDS-gel electrophoresis under reducing conditions showed that the norrin expressed had an apparent Mr = 14,000 and was present only in cell lysates and the extracellular matrix. Under nonreducing conditions, most of the norrin in the extracellular matrix was oligomers that contained up to approximately 20 monomers. One of the major extracellular species of norrin under reducing conditions after cross-linking of norrin oligomers with bis(sulfosuccinimidyl)suberate had an apparent Mr = 28,000, consistent with covalent cross-linked dimers. Thus the covalently cross-linked dimers are key structural components of norrin oligomers. By site-directed mutagenesis, the codon for half-cystine 95 in norrin was changed to one encoding alanine. The norrin C95A found in the extracellular matrix of cells transfected with this mutant was the size of dimers, indicating that half-cystine 95 is involved in oligomer formation. The corresponding half-cystine residue in human prepro-von Willebrand factor is also involved in interchain disulfide bond formation, which is consistent with the sequence identity of the half-cystine residues in norrin and part of the half-cystine residues in a disulfide-rich domain of von Willebrand factor. Replacement of valine at residue 60 in norrin by glutamic acid, a mutation found in humans with a severe type of Norrie disease, results in a considerable reduction (50%) in the amount of norrin in the extracellular matrix of transfected COS-7 cells. Replacement of arginine at residue 121 by glutamine, which is associated with a less severe type of Norrie disease, results in a reduction in the amount of norrin R121Q in the extracellular matrix (26%). These studies suggest that norrin is a secreted protein that forms disulfide bonded oligomers that are associated with the extracellular matrix upon secretion from cells. Moreover, the disulfide-rich motif of norrin and prepro-von Willebrand factor promotes interchain disulfide bond formation among polypeptides in which it is found. PMID- 9407137 TI - Caveolin interaction with protein kinase C. Isoenzyme-dependent regulation of kinase activity by the caveolin scaffolding domain peptide. AB - Caveolar localization of protein kinase C and the regulation of caveolar function by protein kinase C are well known. This study was undertaken to examine whether caveolin subtypes interact with various protein kinase C isoenzymes using the caveolin scaffolding domain peptide. When protein kinase C-alpha, -epsilon, and zeta were overexpressed in COS cells followed by subcellular fractionation using the sucrose gradient method, all the isoenzymes (alpha, epsilon, and zeta) were detected in the same fraction as caveolin. The scaffolding domain peptide of caveolin-1 and -3, but not -2, inhibited the kinase activity and autophosphorylation of protein kinase C-alpha and -zeta, but not of protein kinase C-epsilon, overexpressed in insect cells. Truncation mutation studies of the caveolin-1 and -3 peptides demonstrated that a minimum of 16 or 14 amino acid residues of the peptide were required for the inhibition or direct binding of protein kinase C. Thus, the caveolin peptide physically interacted with protein kinase C and regulated its function. Further, this regulation occurred in a protein kinase C isoenzyme-dependent manner. Our results may provide a new mechanism regarding the regulation of protein kinase C isoenzyme activity and the molecular interaction of protein kinase C with its putative binding proteins. PMID- 9407138 TI - The JUN kinase/stress-activated protein kinase pathway is required for epidermal growth factor stimulation of growth of human A549 lung carcinoma cells. AB - Epidermal growth factor (EGF) plays a major role in non-small cell lung cancer cell autocrine growth and has been reported to activate the JUN kinase/stress activated protein kinase (JNK/SAPK) pathway in model cells. Activation of JNK/SAPK leads to the phosphorylation of c-JUN protooncogene on serines 63 and 73. This mechanism is required for and cooperates in the transformation of rat embryo fibroblasts by Ha-RAS. However, the function of JNK/SAPK in human tumor growth is unknown. We have tested several lung carcinoma cell lines. All exhibited UV-C-inducible JNK/SAPK activity; two exhibited constitutive activity in low serum, and two (M103 and A549) exhibited EGF-inducible JNK/SAPK activity. In A549 cells, EGF induced a rapid and prolonged (up to 24 h) activation of the JNK/SAPK pathway that correlated with a 150-190% growth stimulation. Stably transfected clones of A549 cells expressing c-JUN(S63A,S73A), a transdominant inhibitor of c-JUN, completely blocked the EGF-stimulated proliferation effect but did not alter the basal proliferation rate. Consistent with these results JNK antisense oligonucleotides targeted to JNK1 and JNK2 entirely eliminated the EGF stimulated JNK/SAPK activity and blocked EGF-stimulated growth but not basal growth. In contrast, specific inhibition of the RAF/ERK pathway by PD98059 (MEK1 inhibitor) completely blocked ERK activation by EGF and basal cell growth but not EGF-stimulated growth, thereby dissociating the growth-promoting roles of each pathway. Our observations indicate, for the first time, that JNK/SAPK may be a preferential effector pathway for the growth properties of EGF in A549 cells. PMID- 9407139 TI - Parallel dimers and anti-parallel tetramers formed by epidermal growth factor receptor pathway substrate clone 15. AB - The recently discovered localization of epidermal growth factor receptor pathway substrate clone 15 (Eps15) to plasma membrane clathrin-coated pits and its constitutive association with the endocytic clathrin adaptor protein complex, AP 2, strongly suggest that Eps15 has an important role in the pathway of clathrin dependent endocytic traffic. We report here that Eps15 forms dimers and tetramers of distinct shape. The Eps15 dimer is an elongated molecule, 32 nm in length. There is a globular "head" at one end of the molecule and an extended "stalk" of 25 nm which is kinked at about 17 nm away from the head. In the Eps15 dimer, two subunits are arranged parallel to each other, so that the head corresponds to two side by side copies of the N-terminal region I, which contains the three Eps15 homology domains. The proximal part of the stalk is the coiled-coil central region II containing 20 heptad repeats. The kink is at the boundary between region II and the C-terminal region III, which contains the AP-2 binding site, 15 aspartic-proline-phenylalanine repeats, and proline-rich Src homology domain ligand sites. The Eps15 tetramer has a "dumbbell" shape, approximately 31 nm in length; it is formed by the anti-parallel association of two Eps15 dimers. Formation of these Eps15 tetramers appears to require contacts between regions I of one dimer and regions III of a second apposing dimer. The extended shapes of the Eps15 dimers and tetramers suggest how Eps15 oligomers are located in the clathrin coat. We discuss the implications for accessibility to partners and for proposed functions of Eps15. PMID- 9407141 TI - MPW: the Metabolic Pathways Database. AB - The Metabolic Pathwasy Database (MPW) (www.biobase.com/emphome.html/homepage. html.pags/pathways.html) a derivative of EMP (www.biobase.com/EMP) plays a fundamental role in the technology of metabolic reconstructions from sequenced genomes under the PUMA (www.mcs.anl.gov/home/compbio/PUMA/Production/ ReconstructedMetabolism/reconstruction.html), WIT (www.mcs.anl.gov/home/compbio/WIT/wit.html ) and WIT2 (beauty.isdn.msc.anl.gov/WIT2.pub/CGI/user.cgi) systems. In October 1997, it included some 2800 pathway diagrams covering primary and secondary metabolism, membrane transport, signal transduction pathways, intracellular traffic, translation and transcription. In the current public release of MPW (beauty.isdn.mcs.anl.gov/MPW), the encoding is based on the logical structure of the pathways and is represented by the objects commonly used in electronic circuit design. This facilitates drawing and editing the diagrams and makes possible automation of the basic simulation operations such as deriving stoichiometric matrices, rate laws, and, ultimately, dynamic models of metabolic pathways. Individual pathway diagrams, automatically derived from the original ASCII records, are stored as SGML instances supplemented by relational indices. An auxiliary database of compound names and structures, encoded in the SMILES format, is maintained to unambiguously connect the pathways to the chemical structures of their intermediates. PMID- 9407140 TI - p300 functions as a coactivator for the peroxisome proliferator-activated receptor alpha. AB - The integrator protein, p300, was demonstrated to interact with mouse peroxisome proliferator-activated receptor alpha in a ligand-enhanced manner. The PPARalpha interacting domain of p300 was mapped to amino acids 39-117 which interacted strongly with PPARalpha but did not interact with retinoic acid receptor-gamma or retinoid X receptor-alpha. Amino acids within the carboxyl terminus of PPARalpha as well as residues within the hinge region were required for ligand-dependent interaction with p300. p300 enhanced the transcriptional activation properties of PPARalpha and, therefore, can be considered a bona fide coactivator for this nuclear receptor. These observations extend the group of p300-interacting proteins to include mPPARalpha and further characterize the molecular mechanisms of PPARalpha-mediated transcriptional regulation. PMID- 9407151 TI - Chemokine receptors and human immunodeficiency virus infection. AB - Primate lentiviruses infect target cells by interacting with the cell surface protein, CD4 and additional molecules, termed coreceptors. Recently, HIV-1 coreceptors have been identified as seven transmembrane spanning, G-protein coupled receptors of the chemokine receptor family. Thus, expression of CD4 and an appropriate coreceptor is both necessary and sufficient to render target cell permissive for fusion with virions or infected cells. The spectrum of tissue tropisms exhibited by primate lentiviruses can be largely explained by differential utilization and distribution of coreceptors. This article reviews what is currently known about the selective utilization of particular coreceptors by primate lentiviruses and the nature of the envelope/coreceptor interaction, with particular reference to two important HIV-1 coreceptors, CCR-5 and CXCR-4. It has become clear that these interactions are somewhat 'plastic': Variability is evident, both in the selection of coreceptor and the way in which different viral strains interact with their cognate coreceptors. The implications of these findings both for attempts to block HIV infection with coreceptor targeted agents and for understanding HIV replication in vivo is discussed. PMID- 9407152 TI - How do intracellular proteolytic systems change with age? AB - One of the common features of cells from senescent tissues is the accumulation of abnormal proteins. Several hypotheses have been proposed to explain the origin of those abnormal proteins. A defect in proteolytic systems usually responsible for the elimination of altered proteins from the cells could clearly contribute to such accumulation. Here we describe the effect of age on the major proteolytic systems within cells: the ubiquitin-proteasome pathway, the calcium-activated calpain pathways, and multiple lysosomal pathways. Our group has contributed to the characterization of a selective pathway of degradation of cytosolic proteins in lysosomes that is activated under conditions of nutrient deprivation. In this lysosomal pathway of proteolysis proteins are transported through the lysosomal membrane assisted by cytosolic and lysosomal molecular chaperones and a receptor protein in the lysosomal membrane. The activity of this pathway significantly decreases with age, and this decrease might account for the cytosolic accumulation of aberrant substrate proteins in senescent cells. The cellular consequences of the decline of this lysosomal pathway together with possible methods to restore the reduced function are also addressed in this review. PMID- 9407153 TI - The threshold for prophylactic platelet transfusions in adults with acute myeloid leukemia. Gruppo Italiano Malattie Ematologiche Maligne dell'Adulto. AB - BACKGROUND: Prophylactic platelet transfusions are usually administered to patients receiving myelotoxic chemotherapy when their platelet count falls below 20,000 per cubic millimeter. Some observations suggest that lower platelet counts can be appropriate in patients in stable condition, but the safety of lower thresholds is uncertain. METHODS: We evaluated 255 adolescents and adults (age, 16 to 70 years) with newly diagnosed acute myeloid leukemia (but not acute promyelocytic leukemia), who were treated in 21 centers. One hundred thirty-five patients were randomly assigned to receive a transfusion when their platelet count fell below 10,000 per cubic millimeter (or 10,000 to 20,000 per cubic millimeter in those with a temperature above 38 degrees C, with active bleeding, or a need for invasive procedures), and 120 patients were assigned to receive a transfusion when their platelet count was less than 20,000 per cubic millimeter. RESULTS: Patients in the group with a threshold of 10,000 platelets per cubic millimeter received 21.5 percent fewer platelet transfusions than the patients in the group with a threshold of 20,000 platelets per cubic millimeter (P=0.001). The numbers of red-cell units transfused were not significantly different between groups. Major bleeding (defined as any bleeding more than petechiae or mucosal or retinal bleeding) occurred in 21.5 and 20 percent of patients, respectively (P=0.41), and on 3.1 and 2.0 percent of the days of hospitalization. One episode of fatal cerebral hemorrhage occurred in the group with a threshold of 10,000 platelets per cubic millimeter; none occurred in the other group (P= 0.95). Actuarial estimates of survival during induction chemotherapy, actuarial estimates of the absence of major bleeding, and the length of hospital stay were not significantly different in the two groups. CONCLUSIONS: The risk of major bleeding during induction chemotherapy in adolescents and adults with acute myeloid leukemia (except acute promyelocytic leukemia, which we did not study) was similar with platelet-transfusion thresholds of 20,000 per cubic millimeter and 10,000 per cubic millimeter (or 10,000 to 20,000 per cubic millimeter when body temperature exceeded 38 degrees C, there was active bleeding, or invasive procedures were needed). Use of the lower threshold reduced platelet use by 21.5 percent. PMID- 9407154 TI - Molecular epidemiology of bartonella infections in patients with bacillary angiomatosis-peliosis. AB - BACKGROUND: Bacillary angiomatosis and bacillary peliosis are vascular proliferative manifestations of infection with species of the genus bartonella that occur predominantly in patients infected with the human immunodeficiency virus. Two species, B. henselae and B. quintana, have been associated with bacillary angiomatosis, but culture and speciation are difficult, and there has been little systematic evaluation of the species-specific disease characteristics. We studied 49 patients seen over eight years who were infected with bartonella species identified by molecular techniques and who had clinical lesions consistent with bacillary angiomatosis-peliosis. METHODS: In this case control study, a standardized questionnaire about exposures was administered to patients with bacillary angiomatosis-peliosis and to 96 matched controls. The infecting bartonella species were determined by molecular techniques. RESULTS: Of the 49 patients with bacillary angiomatosis-peliosis, 26 (53 percent) were infected with B. henselae and 23 (47 percent) with B. quintana. Subcutaneous and lytic bone lesions were strongly associated with B. quintana, whereas peliosis hepatis was associated exclusively with B. henselae. Patients with B. henselae infection were identified throughout the study period and were epidemiologically linked to cat and flea exposure (P< or =0.004), whereas those with B. quintana were clustered and were characterized by low income (P=0.003), homelessness (P = 0.004), and exposure to lice (P= 0.03). Prior treatment with macrolide antibiotics appeared to be protective against infection with either species. CONCLUSIONS: B. henselae and B. quintana, the organisms that cause bacillary angiomatosis-peliosis, are associated with different epidemiologic risk factors and with predilections for involvement of different organs. PMID- 9407155 TI - Absence of toxicity of oats in patients with dermatitis herpetiformis. AB - BACKGROUND: People with gluten sensitivity should avoid foods containing wheat, rye, and barley, but there has been debate about whether they should avoid oats. Although patients with celiac disease have recently been shown to tolerate oats, less is known about the effects of oats on patients with dermatitis herpetiformis. METHODS: We studied seven men and three women (mean age, 58 years) with biopsy-confirmed dermatitis herpetiformis. They had followed a strict gluten free diet for a mean of 15.8 years, which controlled their rash and enteropathy. The patients added oats that were not contaminated with gluten to their diets for 12 weeks (mean [+/-SD] daily intake, 62.5+/-10.8 g). RESULTS: None of the patients had any adverse effects. Serologic tests for antigliadin, antireticulin, and antiendomysial antibodies were negative before oats were introduced into the diet and after they were discontinued. Villous architecture remained normal: the mean (+/-SE) ratio of the height of villi to the depth of crypts was 3.59+/-0.11 before the diet and 3.71+/-0.09 afterward (normal, 3 to 5), and the mean enterocyte heights were 31.36+/-0.58 microm and 31.75+/-44 microm, respectively (normal range, 29 to 34). Duodenal intraepithelial lymphocyte counts all remained within normal limits (mean, 13.8+/-1.03 per 100 enterocytes before the diet and 14.2+/-1.2 per 100 enterocytes afterward; normal range, 10 to 30). Dermal IgA showed no significant changes. CONCLUSIONS: Patients with dermatitis herpetiformis can include moderate amounts of oats in their gluten-free diets without deleterious effects to the skin or intestine. PMID- 9407156 TI - Images in clinical medicine. Bacillary angiomatosis or Kaposi's sarcoma? PMID- 9407157 TI - Cumulative impact of sustained economic hardship on physical, cognitive, psychological, and social functioning. AB - BACKGROUND: Although the relation between low income and poor health is well established, most previous research has measured income at only one time. METHODS: We used income information collected in 1965, 1974, and 1983 from a representative sample of adults in Alameda County, California, to examine the cumulative effect of economic hardship (defined as a total household income of less than 200 percent of the federal poverty level) on participants who were alive in 1994. RESULTS: Because of missing information, analyses were based on between 1081 and 1124 participants (median age, 65 years in 1994). After adjustment for age and sex, there were significant graded associations between the number of times income was less than 200 percent of the poverty level (range, 0 to 3) and all measures of functioning examined except social isolation. As compared with subjects without economic hardship, those with economic hardship in 1965, 1974, and 1983 were much more likely to have difficulties with independent activities of daily living (such as cooking, shopping, and managing money) (odds ratio, 3.38; 95 percent confidence interval, 1.49 to 7.64), activities of daily living (such as walking, eating, dressing, and using the toilet) (odds ratio, 3.79; 95 percent confidence interval, 1.32 to 9.81), and clinical depression (odds ratio, 3.24; 95 percent confidence interval, 1.32 to 7.89) in 1994. We found little evidence of reverse causation -- that is, that episodes of illness might have caused subsequent economic hardship. CONCLUSIONS: Sustained economic hardship leads to poorer physical, psychological, and cognitive functioning. PMID- 9407158 TI - Vaginitis. PMID- 9407160 TI - The perils of platelet transfusions. PMID- 9407161 TI - Of cats, humans, and Bartonella. PMID- 9407162 TI - The Medical Repository--the first U.S. medical journal (1797-1824). PMID- 9407163 TI - Telogen effluvium. AB - Most of the patients complaining of hair loss do not fit the diagnosis of androgenetic alopecia (AGA). This paper's objective is to describe acute and chronic telogen effluvium (TE), with common occurrences in both sexes and at any age, by clinical and histopathological observation. Acute and chronic TE is described. A possible pathogenic mechanism and a diagnostic approach are suggested, especially to distinguish chronic TE from AGA. Possible therapies are proposed. The recognition of a chronic variant of TE and its distinction from pure AGA, which very often it overlaps, are of paramount importance for a correct clinical and experimental approach to the problems of hair loss. PMID- 9407164 TI - Increased synergistic effect of EGF and IGF-I on DNA synthesis of cultured psoriatic keratinocytes. AB - BACKGROUND: DNA synthesis is significantly more stimulated in response to insulin like growth factor I (IGF-I) in growth-arrested cultures of psoriatic than of nonpsoriatic keratinocytes. Epidermal growth factor (EGF), by itself only a weak stimulator, was recently found to cause a strong synergetic effect when added together with IGF-I to newborn human keratinocytes. OBJECTIVE: (1) To measure this effect in cultured adult psoriatic and nonpsoriatic keratinocytes, and (2) to examine whether the difference in DNA synthesis after IGF-I addition could be due to differences in the binding of this growth factor. METHODS: 3H deoxythymidine incorporation and 125I-IGF-I binding studies. RESULTS: Psoriatic keratinocytes demonstrate a considerably stronger response to the combination of EGF and IGF-I than nonpsoriatic keratinocytes. This is apparently not caused by differences in binding of IGF-I. CONCLUSION: Differences in the proliferative rate of normal and psoriatic keratinocytes may be in part due to differences in signal transduction distal from the IGF-I receptor or in the cooperation of the EGF and the IGF-I receptor. PMID- 9407165 TI - Skin surface lipids inhibit adherence of Candida albicans to stratum corneum. AB - BACKGROUND: Candida albicans (CA) infections represent a significant threat to the health of immunocompromised individuals. The initial step in the establishment of a CA infection is adherence of the organism to an epithelial surface. METHODS: An in vitro model for studies on adherence of CA to keratinized surfaces has been developed and used to test the hypothesis that specific lipids can modulate adherence of this organism. Porcine stratum corneum (SC) disks were incubated with candidal suspensions, after which unattached cells were washed away. Adherent cells were stained and counted using light microscopy. RESULTS: Attachment of three pathogenic CA isolates was significantly greater than attachment of commensal strains of either CA or Candida parapsilosis. Removal of lipid from the SC lead to a doubling of the number of adherent organisms, whereas additional skin lipid inhibited adherence. Squalene, wax esters, cholesterol esters and triglycerides had no effect on adherence, but fatty acids, sterols and ceramides inhibited adherence significantly. CONCLUSIONS: Specific epithelial lipids can modulate adherence of CA to keratinized epithelial surfaces. PMID- 9407166 TI - Overexpression of p53 protein associated with proliferative activity as evaluated by Ki-67 immunostaining in well-differentiated squamous cell carcinoma of the skin. AB - BACKGROUND: Overexpression of p53 protein has been demonstrated in a variety of human malignant tumors, and its role in oncogenesis and tumor progression is thought to be important. Recently, some reports have suggested that p53 overexpression might be an indicator of immaturity or proliferative activity of cells in cutaneous lesions. OBJECTIVE: The aim of the present study was to clarify the relationship between overexpression of p53 protein and the proliferative activity evaluated by Ki-67 immunostaining in well-differentiated squamous cell carcinoma (SCC) of the skin. METHODS: Sixteen cases of well differentiated SCC were analyzed by immunohistochemistry for the expression of p53 protein and Ki-67 antigen. RESULTS: p53 protein was detected in 12 out of the 16 cases (75%). The percentage of p53-protein-positive tumor cells (DO-7 index) ranged from 0 to 80.4 (mean +/- SD = 33.8 +/- 29.7) and the percentage of Ki-67 antigen-positive tumor cells (MIB-1 index) from 23.4 to 65.1 (mean +/- SD = 42.3 +/- 15.8). Cases with higher DO-7 indices tended to show higher MIB-1 indices and the correlation was statistically significant. CONCLUSION: Overexpression of the p53 protein seems to be a useful indicator of proliferative activity in cutaneous SCC. PMID- 9407167 TI - Schwannomatosis: a clinical entity distinct from neurofibromatosis type 2. AB - BACKGROUND: Schwannomatosis includes multiple cutaneous schwannomas, central nervous system tumors and various neurologic deficits. OBJECTIVE: To specify the clinical presentation of schwannomatosis, excluding neurofibromatosis type 2 (NF2). METHODS: Patients with at least 2 cutaneous schwannomas were evaluated for NF2 criteria. RESULTS: 5 men and 2 women (mean age 54 +/- 18 years) we included. Two cases were familial. Cutaneous schwannomas were tender nodules with onset above the age of 30 years. Four patients had paresthesias. Four patients had hypacusis, but auditory brainstem responses showed no retrocochlear involvement. No vestibular schwannoma or central nervous system tumor was found with magnetic resonance imaging in 6 patients. Spinal imaging performed in 6 patients was normal in 5 and showed a spinal schwannoma in the 6th patient. CONCLUSION: We strictly eliminated the diagnosis of NF2 in our patients. Exclusion of NF2 in suspected schwannomatosis is essential for further molecular genetic studies. PMID- 9407168 TI - Demodex folliculorum in renal transplant patients. AB - BACKGROUND: Demodex folliculorum (DF), which is a resident in human pilosebaceous follicles, has been implicated in infections of patients under cancer chemotherapy and with AIDS. OBJECTIVE: We aimed to determine the influence of immunosuppression on carriage of DF. METHODS: Mite density was compared, of 30 renal transplant (RT) patients under a combination therapy of cyclosporine, azathioprine and prednisolone with that of age- and sex-matched controls. Two samples of standardized skin surface biopsies were taken from each subject. RESULTS: DF was not found in any of the RT patients while a mean density of 0.55 DF/cm2 was present in healthy subjects (Fisher's exact test, p < 0.0001). The relationship between mite density and classic risk factors was not found to be statistically significant (Fisher's exact test, p > 0.05). CONCLUSION: Our findings indicate that there may be other factors than immunosuppressive therapy influencing DF density. PMID- 9407169 TI - Perioral dermatitis in children--clinical presentation, pathogenesis-related factors and response to topical metronidazole. AB - BACKGROUND: Perioral dermatitis, a common skin disorder in young women, is rarely described in children. OBJECTIVE: This study elaborates the clinical features of perioral dermatitis in children as well as possible pathogenetic mechanisms and the response to topical metronidazole. METHODS: Seven children (4 females, 3 males between 4 and 12 years of age) were evaluated and dermatological examination was carried out. Pretreatment with topical corticosteroids was documented. Skin prick test with a panel of six common aeroallergens was performed in all children. All children were screened for gastrointestinal colonization with Candida albicans. Patients were treated with topical metronidazole 1% during the first 2 weeks. From the 3rd week on 2% metronidazole was used. RESULTS: In all but one child topical corticosteroids had been used in the face prior to the first presentation at our outpatient department suggesting a possible pathogenetic role. An association with atopy or intestinal candida colonization was not found. In all children skin lesions resolved after 3-6 months. The children remained free of symptoms over an observation period of 2 years. CONCLUSION: Perioral dermatitis has to be considered as differential diagnosis in children presenting with erythematous papules and papulovesicles in typical locations. Metronidazole proved to be effective and safe in the treatment of perioral dermatitis in children. Atopy and gastrointestinal colonization with C. albicans do not seem to play a role in the pathogenesis of perioral dermatitis. PMID- 9407170 TI - Demodicidosis in immunocompetent young children: report of eight cases. AB - Demodex folliculorum and Demodex brevis are obligate parasites of the human pilosebaceous unit. They are the most common permanent ectoparasites in adults, but their incidence on children's skin is rare. Only few cases of demodicidosis have been reported in children aged below 5 years and most of them were suffering from leukemia or HIV infection. The aim of this study is to describe demodicidosis in young immunocompetent children. The clinical details of 8 healthy children are given. There were 4 males and 4 females aged between 10 months and 5 years referred to us for evaluation of a facial eruption characterized by erythema, papulopustules and variable edema. In 7 of these patients, skin scrapings were performed and in 1 a 4-mm punch biopsy. Numerous D. folliculorum were found in skin scrapings of 7 cases and at a histologic examination of skin biopsy in 1 case. We employed topical metronidazole gel 1% in all patients and we obtained a 100% recovery without relapses after a 1- to 3 year follow-up. In 2 of our cases Demodex infestation had a mild form resembling pityriasis folliculorum and the other cases presented a rosacea-like form. The reason why these young immunocompetent children developed demodicidosis is still under evaluation and investigation. PMID- 9407171 TI - Efficacy and safety of stand-up irradiation cubicles with UVA metal-halide lamps (and a new filter) or UVA fluorescent lamps for photochemotherapy of psoriasis. AB - BACKGROUND: Metal-halide lamps (MHLs) and fluorescent lamps (FLs) are widely employed for PUVA therapy of psoriasis, but they have never been compared in a clinical trial. OBJECTIVE: We studied the irradiance, spectral power distribution, irradiation field and efficacy of two cabinets with standard FLs or MHLs with a new UVA filter. METHODS: The photophysical properties of the lamps were studied with a spectroradiometer. After phototesting, 22 patients with recurrent plaque type psoriasis were treated in a stand-up irradiation cubicle housing 27 standard FLs, and, at recurrence, in a cubicle with 15 MHLs. When indicated, lesions of the legs underwent supplementary exposures with small FL or MHL devices. RESULTS: MHLs had a greater emission in the longer UVA wavelengths than FIs. The MHL cubicle had a greater irradiance and a more uniform output along the vertical axis. Both cubicles were effective in a similar number of exposures. However, FLs were more phototoxic and lower cumulative UVA doses were administered although the total duration of exposures was longer. Eleven patients treated with FLs and 7 patients with MHLs required supplementary treatments of the legs. The number, cumulative UVA dose and total duration of these exposures were significantly greater with FLs. CONCLUSION: Cabinets with MHLs increase the number of patients treated in the same time period and reduce the number of patients needing supplementary treatments of the legs. Cumulative UVA doses are greater with MHLs, but their emission is prevalent in the less erythematogenic and carcinogenic longer UVA wavelengths. PMID- 9407172 TI - Suppression of UV-induced erythema by topical treatment with melatonin (N-acetyl 5-methoxytryptamine). Influence of the application time point. AB - BACKGROUND: In a previous study, we reported a significant and dose-dependent suppression of UV-induced erythema in human skin by a topically applied melatonin preparation. OBJECTIVE: The present double-blind randomized study was designed to examine the influence of the application time point of topical melatonin on this antierythema effect. METHODS: Defined small areas on the lower back of 20 volunteers were treated with 0.6 mg/cm2 melatonin dissolved in a nanocolloid gel carrier either 15 min before or 1, 30 or 240 min after UV irradiation with twice the individual minimal erythema dose delivered by a Multiport Solar UV Simulator (UVA and UVB). The erythemata induced were evaluated by visual scoring and chromametry 24 h after irradiation. RESULTS: Treatment of the skin with melatonin 15 min before UV irradiation proved to almost completely suppress the development of an UV-induced erythema. In contrast, no significant protective effects of melatonin were observed when it was applied after UV irradiation. CONCLUSION: Topically applied melatonin has a clear-cut protective effect against UV-induced erythema. Free radical scavenging of UV-generated hydroxyl radicals and interference with the arachidonic acid metabolism are possible mechanisms of the melatonin action. PMID- 9407174 TI - Effects of soap and detergents on skin surface pH, stratum corneum hydration and fat content in infants. AB - BACKGROUND: In adults the influence of cleansing preparations on the pH, fat content and hydration of the skin is well documented. Studies in newborn and small infants have not been reported. OBJECTIVE: Our study aimed at examining whether similar effects can be ascertained in infants. METHODS: Infants without skin disease, aged 2 weeks to 16 months, entered an open, controlled and randomized study. Ten infants each had skin washed with tap water (control group), liquid detergent (pH 5.5), compact detergent (pH 5.5) or alkaline soap (pH 9.5). The pH, fat content and hydration were measured before and 10 min after cleansing. Findings were statistically evaluated by parametric covariance analysis. RESULTS: The skin pH increased from an average of 6.60 after cleansing in all groups. The smallest increase (+0.19) was observed in the control group, the largest (+0.45) after washing with alkaline soap. After treatment with liquid or compact detergent, the increase of the pH was only 0.09 higher than for the control group. In comparison to the compact and liquid detergents, the alkaline soap group had a significantly higher increase in pH. The fat content (mean starting value: 4.34 micrograms/cm2) decreased after washing in all groups; the smallest effect was observed in the control group (decrease of 0.93 micrograms/cm2), the highest for the alkaline soap group (decrease of 4.81 micrograms/cm2). In comparison to the compact and liquid detergents, the alkaline soap group had a higher decrease in fat content. This difference was significant for compact detergents. No statistically significant differences were observed for hydration before versus after washing. CONCLUSION: Each cleansing agent, even normal tap water, influences the skin surface. The increase of the skin pH irritates the physiological protective 'acid mantle', changes the composition of the cutaneous bacterial flora and the activity of enzymes in the upper epidermis, which have an acid pH optimum. The dissolution of fat from the skin surface may influence the hydration status leading to a dry and squamous skin. PMID- 9407173 TI - Application of lipid microspheres containing prostaglandin E1 ointment to peripheral ischemic ulcers. AB - BACKGROUND: The systemic use of prostaglandin E1 (PGE1) in the treatment of peripheral vascular disease is well documented. It is known that the liposomal formulation of some drugs enhances their transdermal absorption. OBJECTIVE: The potential of topical application of lipid microspheres containing PGE1 (lipo PGE1) to treat ischemic ulcers was evaluated. METHODS: Lipo-PGE1 ointment (1 microgram/g) was applied topically to peripheral ischemic ulcers in 10 patients for 5.8 weeks (range 4-9 weeks). The patients were followed up for 6 months, and response was assessed comparing photographs of the lesions. RESULTS: Nine of 10 patients responded to treatment, but in 3 patients, the ulcers recurred after cessation of treatment. CONCLUSION: It is concluded that lipo-PGE1 ointment provides an alternative to the management of patients with incurable peripheral ischemic ulcers. PMID- 9407175 TI - Sclerotic fibroma of tendon sheath. AB - Two cases of sclerotic fibromas directly related to the tendon sheath are presented. Fibroma in these 2 cases consisted of hypocellular, homogeneous collagen bundles that were closely related to the subjacent tendon sheath, and were described by the term 'sclerotic fibroma of tendon sheath'. It is possible that the sclerotic fibroma may arise from fibroma of tendon sheath and that common pathomechanisms exist between these two types of fibroma. PMID- 9407176 TI - Nonscarring alopecia associated with solitary circumscribed neuroma. AB - We report an unusual case of nonscarring alopecia clinically resembling patchy alopecia areata around central solitary circumscribed neuroma. Prompt and spontaneous complete hair regrowth was observed after removal of the tumor. Further knowledge of growth factor and receptor interaction in regulating the hair follicle cycle, as well as on the production of growth factors and cytokines by tumor cells and/or cells within the tumor microenvironment may contribute to better understand the pathologic mechanisms underlying certain curious phenomena such as peritumoral nonscarring alopecia. PMID- 9407177 TI - Staphylococcal scalded skin syndrome in an adult. Identification of exfoliative toxin A and B genes by polymerase chain reaction. AB - A 68-year-old man developed areas of erythema with flaccid bullae and wrinkling on his face, upper extremities, chest, right buttock, and shins. Histological examination revealed an intraepidermal cleavage with a sparse cellular infiltration in the dermis, consistent with a diagnosis of staphylococcal scalded skin syndrome. Cultures from an abscess of his right knee joint and his blood were both positive for Staphylococcus aureus. Both S. aureus isolates were confirmed to be double-producers of ETA and ETB using the polymerase chain reaction. The use of nonsteroidal anti-inflammatory drug seemed to be one of the predisposing factors in this patient. PMID- 9407178 TI - Vasopressin-induced amber-like skin necrosis. AB - Case reports about vasopressin-induced cutaneous necrosis are not frequent. Here we report a further case, of which skin manifestations included not only mottling, cyanosis, ecchymosis, bullae and gangrene, but also amber-like change in focal areas. Besides, intermittent paling of the skin with or without deep pain sensation of the limbs over non-injection sites was observed that might be a warning sign of impending skin necrosis. Based on the literature about vasopressin-induced skin necrosis we discuss the possible role of coagulation enhancement of this molecule. PMID- 9407179 TI - Severe self-healing nail dystrophy in a patient on peritoneal dialysis. AB - The unusual case of a patient on peritoneal dialysis who progressively developed onychodystrophy of the hands 2 months after Acinetobacter peritonitis had been reported. The nail disease consisted of acute pseudoclubbing, elkonyxis, severe Beau's lines and onychomadesis and showed spontaneous recovery within 4 months. Although reminiscent of severe Beau's lines and reflex sympathetic dystrophy, this observation shows a peculiar nail disorder of unknown cause. PMID- 9407180 TI - Scar sarcoidosis in a patient with Crohn's disease. AB - A patient with scar sarcoidosis and Crohn's disease is described. The possibility of patients with Crohn's disease being more prone to develop sarcoidosis is discussed. PMID- 9407181 TI - Focal subungual warty dyskeratoma. AB - Warty dyskeratoma is described in an unusual location. Clinically it appeared as a longitudinal reddish ridge bordered on each side by brown splinter haemorrhages forming a nearly continuous line. The free margin of the nail plate was slightly fissured distal to the red band. In the distal groove, histology showed deep epithelial digitations which lined a crater-like area. Numerous multinucleate giant cells, marked dyskeratosis and acantholytic suprabasal clefts were visible inside this tumour resembling Darier's nail disease. PMID- 9407182 TI - Lupus like lesions in a patient with X-linked chronic granulomatous disease and recombinant X chromosome. AB - Lupus-like lesions in X-linked chronic granulomatous disease (X-CGD) are rare. To our knowledge, only 2 cases have previously been published. We report a 2.5-year old boy with X-CGD whose clinical findings were consistent with cutaneous lupus erythematosus. Conventional histopathology showed epidermal atrophy, parakeratosis, follicular plugging and areas of hydropic degeneration. The most striking feature was a neutrophilic interstitial infiltrate with leukocytoclasia in the upper dermis. The X chromosome of our patient--studied with 2 endonucleases (PstI and TaqI) and 5 probes (P99.6, pERT 87.8, pERT87.15, XJ1.1 and 754)--was recombinant, but we believe that this is an incidental finding, not related to the disease. Neutrophilic infiltrate and leukocytoclasia could be characteristic histopathologic findings of lupus-like lesions in these patients. PMID- 9407183 TI - Isolated lichen planus of the lip successfully treated with chloroquine phosphate. AB - Solitary lesions of lichen planus (LP) are often confused with other inflammatory or malignant epithelial lesions. We describe a 51-year-old woman who had had an LP of the lower lip for 11 years. She had undergone several oral and topical therapies with little improvement. The patient had an excellent response to chloroquine phosphate within 3 months. This report represents the third case of LP exclusively localized on the lower lip. PMID- 9407184 TI - Lichen amyloidosus universalis associated with long-term drug intake. AB - We report a 61-year-old man with hypermelanosis, marked diffuse lichenification and cutaneous lichen amyloidosus. The amyloidosis was attributed to drug intake. PMID- 9407185 TI - Lymphoblastic lymphoma of the pre-B phenotype with cutaneous presentation. AB - Lymphoblastic lymphoma (LBL) is a neoplasm of lymphoid precursors presenting usually as acute leukemia with bone marrow and peripheral blood involvement. Primary cutaneous involvement of LBL with a pre-B phenotype has to be considered an extremely uncommon occurrence, accounting for less than 1% of all non-Hodgkin lymphomas. A child with an LBL involving a single cutaneous manifestation of 6 months duration is presented. At the time of presentation, the lesion consisted of a rapidly enlarging deeply infiltrated tumor on the upper arm. Immunophenotypic analysis performed an paraffin-embedded and frozen tissue sections revealed 2 pre-B phenotype of the tumor cells. Similar results were obtained from lymph node and bone marrow biopsy specimens. After 26 months of polychemotherapy, the patient is currently in complete remission. We wish to add this case to the current literature of LBL with cutaneous involvement, emphasizing the importance of a correct diagnosis and the excellent response to the therapeutic regimen. PMID- 9407186 TI - Polymerase chain reaction and reverse cross blot hybridization assay for detection of mycobacterial DNA in lupus vulgaris. AB - For 5 years, an 83-year-old man had been suffering from slightly itchy erythematous plaques with clearcut margins, located on his left thigh and on his right arm; in addition, on his right auricle there was an erythematous patch with yellowish shadings that had appeared about 3 years before and had progressively spread to the temporal-zygomatic region, the chin and the mandibular arch. These lesions were strongly suggestive of lupus vulgaris; however the conventional bacteriological examinations performed on the biopsy specimen from lesional skin were negative. A diagnosis of lupus vulgaris was achieved through the detection of the 16S rRNA gene of Mycobacterium tuberculosis in a skin biopsy of the patient by means of a polymerase chain reaction followed by a reverse cross blot hybridization, a method which allows the identification of different mycobacterial species in a single hybridization procedure. PMID- 9407187 TI - Overexcretion of low-sulphated chondroitin sulphate in the urine of the patient resembling progeroid. AB - We report an unusual case with mental retardation, short stature, sparse scalp hair, prominence of scalp veins, atrophy of subcutaneous fat, pterygia of the neck and loose skin. The patient excreted greater amounts of low-sulphated chondroitin sulphate (LSC) in the urine than age-matched controls. The pattern of glycosaminoglycan in serum and its synthesis by the patient fibroblasts were normal. Collagen, elastin and decorin mRNA levels in the patient fibroblasts were also unaltered. These results suggest that this patient seems to be different from Lowe's syndrome and decorin-deficient progeroid. An abnormal LSC metabolism may be partially responsible for the pathology of these syndromes. PMID- 9407189 TI - Iontophoretic patch test in allergic contact dermatitis: 30 years' experience. PMID- 9407188 TI - Early ritonavir-induced maculopapular eruption. AB - We report 2 cases of maculopapular eruption and fever in patients infected with human immunodeficiency virus (HIV) on the 2nd day of first administration of ritonavir, a protease inhibitor. In the 1st patient, clinical improvement occurred despite continuation of therapy, and in the 2nd the treatment was stopped with remission and rechallenge resulting in recurrence. Ritonavir should be added to the list of drugs that can induce adverse cutaneous reactions in HIV patients. PMID- 9407190 TI - Disseminated superficial porokeratosis: an eruptive pruritic papular variant. PMID- 9407191 TI - Diffuse hair loss following multiple honeybee stings. PMID- 9407192 TI - Syndrome resembling systemic lupus erythematosus induced by carbamazepine. PMID- 9407193 TI - Lamotrigine and toxic epidermal necrolysis. PMID- 9407194 TI - Amended reports: when, if ever? PMID- 9407195 TI - Something old and nothing new. PMID- 9407196 TI - Cytotechnologist productivity standards: monitoring performance in a TQM environment. PMID- 9407197 TI - Strife in the fast lane. PMID- 9407198 TI - Cytotechnology educators' forum: what happened and where are we going? PMID- 9407199 TI - Is reorganization in your future? PMID- 9407200 TI - Cytological pitfalls in bronchopulmonary tumors. PMID- 9407201 TI - Challenges in the interpretation of FNAs from the salivary glands. PMID- 9407202 TI - Diagnostic problems in thyroid FNAs. AB - The use of FNA cytology to diagnose pathologic conditions of the thyroid has increased considerably in recent years, particularly since it has reduced by half the number of patients undergoing surgery. On the one hand, this diagnostic technique has attracted a certain amount of well justified criticism, but on the other, recent cytohistologic correlations and new scientific knowledge are continually improving its application. We shall discuss the latter aspect in more detail and deal with some simple but informative points which the pathologist may find useful in daily practice. PMID- 9407203 TI - Role of fine-needle aspiration in clinical management of transplant patients. AB - Fine-needle aspiration (FNA) of superficial and deep seated lesions has been used with high sensitivity and specificity in the diagnosis of neoplastic and non neoplastic entities. However, studies of FNA in post-transplant patients are virtually absent. Six hundred and seventy-four allograft recipients (cardiac 288, renal 250, lung 131 and heart-lung 5) were reviewed. A total of 30 (25 heart, 4 lungs and 1 renal transplant) patients underwent an FNA procedure. There were 26 males and 4 females. Ages ranged from 18-63 yr (mean 48 yr). The most common entity aspirated was post-transplant lymphoproliferative disorder (PTLD) in 12 cases, followed by inflammatory lesions in 10 cases, malignant epithelial neoplasms in 3 cases, and 1 case each of malignant mesenchymal tumor, pulmonary infarction, hamartoma of liver, fatty changes of liver, and a benign vascular lesion. Surgical or autopsy tissue was available in 19 cases (63.3%). There was an agreement between tissue diagnosis and FNA material in 18 cases (94.7%). One (5.2%) false negative case was recorded. This was a liver aspirate showing benign liver elements, which a surgical biopsy proved to be a bile duct hamartoma. No false positive cases were recorded. FNA is a highly sensitive and specific diagnostic tool in the management of post-transplant patients. PMID- 9407204 TI - Does p53 immunostaining improve diagnostic accuracy in urine cytology? AB - The frequent change of the transitional cell carcinoma of the urinary tract accounts for the fact that cytological abnormalities in urinary specimens are often not sufficient to enable a definitive diagnosis of malignancy. The purpose of this work was to evaluate the possible use of p53 protein in increasing the diagnostic accuracy of urinary cytology. The expression of p53 was investigated by immunocytochemistry in two groups of urinary specimens, one cytologically positive and the other cytologically negative for cancer. Immunostaining was carried out using a monoclonal antibody to p53. In the positive group, in which bladder cancer was confirmed by cystoscopy and biopsy (31 cases), positive reaction for p53 was found in 55% of the cases (17 cases). In the negative group (92 cases), presence of cancer was histologically ascertained in 64 cases and in this group 15 cases (23.4%) showed positive p53 staining. In the remaining 28 cases of this group, where TCC was not present, 7 cases showed p53 positivity in non-neoplastic urothelial cells. This result shows that, while immunocytochemical detection of p53 in urinary specimens may be used for prognostic evaluation of patients with bladder cancer, it does not contribute to the diagnostic accuracy in cases with morphologically inconclusive or negative cytology. The sensitivity and specificity of the method in detecting bladder carcinoma were 23.5 and 75%, respectively. PMID- 9407205 TI - Postprocedural Pap Smears: a LEEP of faith? AB - Loop electrosurgical excision procedure (LEEP) is gaining in popularity in the United States as an outpatient alternative to the diagnosis, and potentially the treatment, of cervical intraepithelial neoplasia (CIN). LEEP is fast, simple, performed under local anesthesia, readily learned, and without significant morbidity. As cytopathologists and cytotechnologists, immediate cytologic evaluation of cervico-vaginal smears following LEEP is not the routine; however, there are very specific artifacts, most of which are related to the transfer of thermal energy, which result from the procedure. It is important to recognize these cytomorphologic features for accurate interpretation. The indications and contraindications for LEEP are similar to those for other ablative or excisional procedures. There appears to sacrifice in the efficacy of diagnosing and treating CIN by this method. Factors predictive of disease clearance are as confounding as they are for any other cone procedure. At the University of Iowa Hospital and Clinics (UIHC), immediate post-LEEP endocervical brush (PLEB) is often performed as a method of assessing the endocervical canal for residual disease or skip lesions. The most common cytomorphologic features observed are: "taffy-pulled" nuclei in elongated endocervical cells; cell aggregates with coalesced cytoplasm; hockey stick nuclei; notched and enlarged nuclei; and, smudgy chromatin. The difficulties or most frequent diagnostic dilemmas in interpreting these smears initially include abundant blood and smudgy chromatin, often tempting an interpretation of "unsatisfactory". However, careful study reveals that these changes are related to the nature of the procedure and reproducible. Recognition and familiarization of these features enables more accurate interpretation of PLEB cytology. The significance of abnormal PLEB, with regard to disease clearance, is still uncertain. PMID- 9407206 TI - Cytologic correlates of benign versus dysplastic abnormal keratinization. AB - The purpose of this study was to determine the cytologic and histologic features that differentiate benign from squamous intraepithelial lesion (SIL)-associated cervical abnormal keratinization, defined as hyperkeratosis, parakeratosis, or individual cell dyskeratosis. Fifty-four cervical Papanicoloau (Pap) smears that contained abnormally keratinized cells were reviewed without knowledge of the concurrent biopsy. Twenty-three Pap smears were diagnosed as SIL and the corresponding biopsy showed SIL in 21 (91%) of these cases. Of the 23 Pap smears diagnosed as negative for SIL, the corresponding biopsy in 20 cases (87%) showed benign (SIL negative) abnormal keratinization. Eight Pap smears showed squamous atypia, of these 5 showed SIL on biopsy, and the other 3 revealed benign keratinization. The Pap smear correlates of the 25 biopsies that were negative for SIL included marked hyperkeratosis (18/25-72 vs. 5/29-17% for biopsies with SIL) and regular nuclear membranes (16/18-89% cases with nucleated dyskeratotic cells vs. 5/29-17% for biopsies with SIL). The cytologic correlates of the 29 biopsies that showed SIL included irregular chromatin clumping (27/29-93% vs. 3/18-17% for biopsies without SIL), and a disorganized growth pattern (24/29-83 vs. 5/25-20% for biopsies without SIL). It is concluded that the cytologic distinction between benign and SIL-related Pap smears with abnormal keratinization can be reliably made by the degree of hyperkeratosis, nuclear chromaticity pattern and contour, and the growth pattern of the dyskeratotic cells. PMID- 9407207 TI - Seasonal fluctuations in the cervical smear detection rates for (pre)malignant changes and for infections. AB - The detection of diseases can exhibit seasonal fluctuations. This can be studied in cervical smears. Over a 9-year observation span (January 1983-January 1992) a series of 504,093 cervical smears obtained from a routine cytology laboratory in The Netherlands were examined for infections (monilia, trichomonas, actinomyces, human papilloma virus [HPV], chlamydia, and herpes) as well as for mild, moderate, and severe dysplasias, carcinoma in situ, and squamous carcinoma. Statistical analysis (principal component analysis) demonstrates clear seasonal rhythms in the detection of infections as well as in precursor lesions. These findings suggest that we are dealing with "true" detection rhythms. For the detection of (pre)malignancy and HPV, yearly fluctuations in women being screened might be the explanation for our observations. PMID- 9407208 TI - Evaluation of human conjunctival epithelium by a combination of brush cytology and flow cytometry: an approach to the quantitative technique. AB - Cytology using the brush technique is readily available and is a rapid means of establishing a presumptive diagnosis of ocular surface changes. However, those techniques have some limitations when obtaining cells from certain localized areas particularly if using the Cytobrush-S. We have described here a new type of brush (Accellon-M), which can collect the local cells using its spherical tip, and evaluated them by comparing with Cytobrush-S. Furthermore, we differentiated epithelial cells from nonepithelial cells that were collected by brushing, and by a combination of brush cytology and flow cytometry using an anti-keratin antibody, AE-3, which reacts with all basic epithelial keratins. Accellon-M could collect the epithelial cells from conjunctiva as effectively as with the Cytobrush-S, and there were no statistical differences between both groups. AE-3, which is reported as the marker of epithelial cells, were detected quantitatively by a combination of brush cytology and flow cytometry techniques. The result of the present study emphasizes that both the Cytobrush-S and the Accellon-M are valuable for conjunctival brush cytology. An additional positive feature of the Accellon-M may be improved cell collection from the conjunctival epithelium especially when the target cells are in a limited area. The combination of the brush cytology and the flow cytometry technique appears to be a useful adjunct as an additional diagnostic or research tool for use in the detection of various antigens in the conjunctival epithelium. PMID- 9407210 TI - Lymphoid cell aggregates: a useful clue in the fine-needle aspiration diagnosis of follicular lymphomas. AB - Among the various types of lymphoma, follicular lymphoma (FL) is known to have significant limitations in cytologic diagnosis by the fine-needle aspiration (FNA) method. The diagnostic accuracy (DA) for non-Hodgkin's lymphoma (NHL) by FNA was evaluated by review of 82 cases of histologically proved NHL after prior FNA. The DA for all NHLs was 66% (54/82), and that for low-grade lymphomas, including small lymphocytic lymphoma, follicular small-cleaved cell lymphoma, and follicular mixed cell lymphoma, was 71% (12/17). The DA for FL was 69% (11/16). Review of individual surgical and cytologic materials from FLs revealed a tendency to show fibrosis in the cytologically false-negative group and diffuse areas of lymphoma in the true-positive group. The presence of "aggregation" of uniform lymphoid cells, probably due to cell adhesions with the support of dendritic reticulum cells, was seen in 55% of true-positive FL (6/11). In contrast, only 28% of true-positive diffuse large cell lymphomas (5/18) showed a mild degree of aggregation, and none of 7 cases of true-positive diffuse small cleaved cell lymphoma showed this feature. The aggregation of cells was not pathognomonic of FL, but its presence with a homogeneous cellular constituent and the paucity of tingible-body macrophages helped us to predict FL. Also, it was a feature distinguishing FL from diffuse small-cleaved cell lymphoma (P = 0.025). PMID- 9407209 TI - Accuracy of grading gliomas on CT-guided stereotactic biopsies: a survival analysis. AB - The accuracy of using a combination of cytopathologic and histopathologic techniques to diagnose stereotactically guided brain biopsies was investigated in 74 patients. Diagnostic accuracy was assessed by determining whether classification of the biopsies as gliosis, astrocytoma (A), anaplastic astrocytoma (AA), or glioblastoma multiforme (GBM) predicted survival. The utility of on-site evaluation using Diff-Quik-stained crush preparations was also assessed. The patients ranged in age from 5 to 88 years (mean, 55 years) and were followed for over 2 years in most cases. Four cases (5%) were classified as gliosis (G), 7 (9%) as atypical gliosis (AG), 4 (5%) as high-grade mixed oligodendroglioma/astrocytoma (OA), 11 (15%) as astrocytoma (A), 21 (28%) as anaplastic astrocytoma (AA), and 27 (36%) as glioblastoma multiforme (GBM). Median survival was 11 months in patient with OA, 57 months in patients with A, 10 months in patients with AA, and 5 months in patients with GBM. Diagnosis of Diff-Quik-stained crush preparations made during the biopsy procedure was highly correlated with the final diagnosis and survival. We conclude that the diagnosis of stereotactic brain biopsies using cytopathology with on-site evaluation in combination with histopathological evaluation of needle cores is accurate based on a survival analysis. However, A and G may be difficult to distinguish. PMID- 9407211 TI - Local anesthesia for fine-needle aspiration biopsy of palpable breast masses: the effectiveness of a jet injection system. AB - To determine the effectiveness of the Biojector 2000 needle-free lidocaine injection system in achieving satisfactory local anesthesia for fine-needle aspiration (FNA) of palpable breast lesions, we studied 29 female patients. Each patient served as her own control and had two FNA biopsies performed on the lesion. The first FNA biopsy was preceded by either no anesthesia, ethyl chloride cold spray, or traditional needle lidocaine injection. The second FNA was preceded by the Biojector 2000. Twenty-four patients (83%) reported that they preferred the Biojector 2000 over either no anesthesia, ethyl chloride spray, or needle and syringe lidocaine injection. The Biojector 2000 needle-free injection system is an effective and useful method of local anesthesia for FNA of palpable breast masses. PMID- 9407212 TI - Syncytial variant of nodular sclerosing Hodgkin's disease: fine-needle aspiration findings in two cases. AB - The syncytial variant is a recently described, uncommon form of nodular sclerosing Hodgkin's disease that was previously termed "sarcomatoid." In addition to foci of typical sclerosis, it is characterized histologically by sheets or clusters of mononuclear Reed-Sternberg variants. These may be arranged around areas of necrosis with variable numbers of neutrophils. In excised material, differential diagnostic considerations include non-Hodgkin's malignant lymphoma, granulocytic sarcoma, malignant melanoma, metastatic carcinoma, thymoma, and metastatic germ cell tumor. We describe the fine-needle aspiration cytologic finding in two examples of this entity. Cohesive clusters and sheets of malignant cells with clear cytoplasm, vesicular nuclei, and prominent nucleoli are easily mistaken for metastatic carcinoma or germ cell tumor. Ancillary tests useful in this differential diagnosis are discussed. PMID- 9407213 TI - Prostatic duct adenocarcinoma: a cytologic and histologic case report with review of the literature. AB - Prostatic duct adenocarcinoma is a rare tumor which typically involves the prostatic urethra as its primary site. Due to this unique location, prostatic duct adenocarcinoma may shed cells into urine specimens which may be detected by cytologic techniques. This report describes the cytologic and histologic features of a case of prostatic duct adenocarcinoma involving the prostatic urethra. Key features which differentiate this neoplasm from the more common transitional carcinomas and the potentially confounding prostatic acinar adenocarcinomas include epithelial clusters with prominent nuclear overlap and nuclear grooves. A comparison to the five previously reported cases is presented. PMID- 9407214 TI - Aneuploid nucleomegaly of bronchial cells in ataxia-telangiectasia: cytologic recognition in bronchial brushings. AB - Ataxia-telangiectasia (AT) is an autosomal recessive disorder of childhood onset characterized by cerebellar ataxia and cutaneous and conjunctival telangiectasias, which affects many systems and organs. One histologic feature of AT is the presence of enlarged dystrophic nuclei, predominantly in satellite cells of sympathetic ganglia and dorsal roots. This paper describes the recognition of nucleomegaly of respiratory cells in bronchial brushings of a 9 year-old patient with AT. The enlarged nuclei displayed smooth nuclear contour, coarse and clumped chromatin granules, and one or two conspicuous nucleoli. The average size was 0.1015 mm in the AT case and 0.0573 mm in control cells. Ploidy analysis demonstrated an aneuploid population of cells with a DNA index of 1.31 and a S-G2M fase of 4.48% in the AT, while the control nuclei showed normal diploid values. To our knowledge, this is the first report of a description of aneuploid nucleomegaly of bronchial cells detected in bronchial smears from a patient with AT. Given that malignant transformations are usually preceded by ploidy alterations, it seems likely that the presence of an aneuploid cell population probably correlates with the increased cancer risk observed in AT patients. Cytopathologists must bear in mind these morphologic features of aneuploid nucleomegaly exhibited by certain cell populations when examining a smear from AT patients. Moreover, this finding may even represent a clue for diagnosis of AT in cases in which the disease has gone unrecognized. PMID- 9407216 TI - Mistakes in cytopathology. PMID- 9407215 TI - Fine-needle aspiration in metastatic glioblastoma multiforme. PMID- 9407217 TI - Identification of mycobacteria by nonradioisotopic single-strand conformation polymorphism analysis. AB - Clinical isolates of mycobacteria were identified to species levels using nonradioisotopic single-strand conformation polymorphism (non-RI SSCP) analysis of 16S rRNA gene fragments amplified by polymerase chain reaction with primers common to all of mycobacterial species. The method is based on a hypervariable region within the 16S rRNA in mycobacteria, which is characterized by species specific nucleotide sequences. A total of 92 mycobacterial strains (Mycobacterium tuberculosis, M. avium, M. gordonae, M. intracellulare, M. kansasii, M. chelonae, M. nonchromogenicum, M. xenopi, and unidentified strain) were studied. They were classified into nine types of pattern showing single-strand DNA bands having different mobilities. Each strain was shown in the species-specific mobility by non-RI SSCP analysis. The results of non-RI SSCP analysis were identical to those of standard biochemical methods and 16S rRNA sequencing. PMID- 9407218 TI - Saccharomyces cerevisiae infections and antifungal susceptibility studies by colorimetric and broth macrodilution methods. AB - Saccharomyces cerevisiae was isolated in large numbers from operative specimens from two patients with perforated bowel and peritonitis and from the blood of another patient treated with extracorporeal membrane oxygenation. Susceptibility studies were performed on these three isolates and another 29 isolates that colonized or caused infection in a total of 19 patients seen over the last decade. All isolates had low minimum inhibitory concentration (MIC) values for amphotericin B (MIC90 of < or = 0.02 microgram/ml) and flucytosine (MIC90 of 0.2 microgram/ml), and a broader range of MIC values for itraconazole (MIC90 of 0.8 microgram/ml) and fluconazole (MIC90 of 4 micrograms/ml). A colorimetric method using Alamar blue reagent showed good concordance with the standard broth macrodilution method for amphotericin B, flucytosine, and fluconazole, but less good concordance for itraconazole. Serious infections with S. cerevisiae probably should be treated with amphotericin B, with or without the addition of flucytosine. PMID- 9407219 TI - Value of the agar plate method for the diagnosis of intestinal strongyloidiasis. AB - An agar plate method for the diagnosis of intestinal strongyloidiasis was compared to the standard formalin-ethyl acetate concentration method. A total of 13 of 225 patients with eosinophilia had positive stools for strongyloides larva by agar plate compared to six of 225 by the formalin-ethyl acetate method (P = .0455). Nine positive stool specimens by the agar plate method were tested by the Baermann technique, and five were positive. The agar plate method is a sensitive and efficient technique for the diagnosis of strongyloidiasis. PMID- 9407220 TI - Macrolides resistance of common bacteria isolated from Taiwan. AB - To determine the susceptibility to macrolides of common pathogenic bacteria isolated from Taiwan, the in vitro activities of erythromycin, roxithromycin, azithromycin, clarithromycin, and dirithromycin were tested against 492 clinical isolates of eight different bacteria, collected from the National Taiwan University Hospital. The results showed high minimum inhibitory concentrations (MICs) against most of the tested bacteria. The MIC90s for Staphylococcus aureus (both methicillin-resistant and -sensitive strains), coagulase-negative staphylococci (both methicillin-resistant and -sensitive strains), Streptococcus pyogenes, Streptococcus pneumoniae, enterococci, peptostreptococci, and Bacteroides fragilis were all > or = 256 micrograms/ml. The MIC50s for methicillin-resistant strains of S. aureus and coagulase-negative staphylococci, and enterococci were > or = 256 micrograms/ml. For S. pneumoniae, peptostreptococci, and B. fragilis, the MIC50s were > 8 micrograms/ml. The resistance rates to macrolides were 80% or more in methicillin-resistant staphylococci and about 30% in methicillin-sensitive staphylococci. Around 55% of S. pneumoniae strains and 37 approximately 42% of S. pyogenes strains were resistant to macrolides. Cross-resistance to different macrolides was clearly demonstrated in most of the resistant strains. PMID- 9407221 TI - Neonatal diarrhea caused by Vibrio cholerae 0139 Bengal. AB - Cholera rarely occurs in children under 2 years of age. We describe diarrhea due to Vibrio cholerae 0139 Bengal, the newly described etiologic agent of cholera in a 4-day-old breast-fed baby. However, the diarrhea was mild and was successfully treated with rehydration therapy and erythromycin. PMID- 9407222 TI - Preliminary interpretive criteria for disk diffusion susceptibility testing of SCH 27899, a compound in the everninomicin class of antimicrobial agents. AB - As antimicrobial resistance among Gram-positive species becomes more common, alternative agents need to be developed for the therapy of serious infections. SCH 27899 is a compound from the everninomicin class of antimicrobial agents that possesses a potent Gram-positive spectrum. We evaluated three disk concentrations (0.25, 1, and 5 micrograms) of three SCH 27899 formulations including SCH 27899 base (SCHB), N-methylglucamine SCH 27899 (NMG-SCH), and NMG-SCH complexed with hydroxypropyl beta-cyclodextrin. Disk zone diameters were correlated with minimum inhibitory concentration for 209 aerobic, nonfastidious Gram-positive strains and selected Gram-negative bacilli to develop disk diffusion interpretive criteria. No significant differences in activity were noted among the three SCH 27899 preparations. Of the three disk concentrations, the correlation coefficient was greatest (r = 0.88) for the 5-micrograms SCHB disk test. For a tentative break point of < or = 2 micrograms SCHB/ml, preliminary disk interpretive criteria were: susceptible at > or = 12 mm, intermediate at 10-11 mm, and resistant at < or = 9 mm (absolute categorical agreement, 99.5%). Zones were small secondary to drug solubility and diffusion limitations. Using these criteria for the SCHB 5 micrograms disks, nearly all of the tested Gram-positive organisms were susceptible including methicillin-resistant staphylococci and vancomycin resistant enterococci. PMID- 9407223 TI - Human papillomavirus detection in high-grade squamous intraepithelial lesions. Comparison of hybrid capture assay with a polymerase chain reaction system. AB - The validity of human papillomavirus (HPV) detection using the hybrid capture assay (HCA) was compared with the polymerase chain reaction (PCR) in 38 patients with high-grade squamous intraepithelial lesions (HSILs). HCA and PCR showed 84% agreement for HPV detection. HCA missed a significant higher proportion of HSIL compared with PCR (21% vs. 5%; P = .04). Thus, the sensitivity of HCA should be increased before this test can be recommended for HSIL. PMID- 9407224 TI - The choroid: from the architect gene to apoptosis. Lecture on acceptance of the Hermann Wacker Prize 1996. PMID- 9407226 TI - Ex vivo phosphorus magnetic resonance spectroscopy on eye bank corneas and corneal metabolic health. AB - BACKGROUND: Since a potential exists for untoward effects on the cornea from the high magnetic fields and radio-frequency energies, and the further manipulation required for phosphorus-31 magnetic resonance spectroscopy (31P-MRS), we determined the effects of this technology on tissues using paired human corneas (n = 4) meeting criteria acceptable for transplantation. METHODS: Slit-lamp biomicroscopy, pachometry, specular microscopy, and redux fluorophotometry were performed on all corneas. One cornea of each pair was examined (< 30 min) by 31P MRS. Following 31P-MRS, slit-lamp biomicroscopy, pachometry, and redox fluorophotometry were again performed. RESULTS: Data tabulated included the 31P energy modulus (1.37 +/- 0.28), the ATP/Pi (2.92 +/- 0.59) and SP/Pi (0.76 +/- 0.04) ratios, and the intracorneal pH (7.24 +/- 0.09). CONCLUSION: Since there were no significant differences in slit-lamp biomicroscopy, endothelial density and morphometry, cell counts, and pachometric and redox fluorophotometric measurements between corneas of each pair before and after 31P-MRS analysis, it was concluded that there was no detectable metabolic damage secondary to such analysis. This study suggests that MRS analysis of human eye-bank tissues does not damage the cornea metabolically and may provide a practical evaluation of the health of the cornea at the biochemical level. PMID- 9407225 TI - Comparison of preferential looking acuity and pattern reversal visual evoked response acuity in pediatric patients. AB - BACKGROUND: We compared the visual acuities obtained with preferential looking (PL), the most widely used method of pediatric vision assessment, with those obtained with the spatial frequency sweep pattern-reversal visual evoked response (SPVER). METHODS: Eighty patients (ages 1.5 months to 12 years) with various ocular pathologies participated in this study. The PL acuity was determined using the up-and-down staircase procedure. The PVER was recorded with the spatial frequency sweep method using 10 spatial frequencies; the acuity was determined by placing the best-fit regression line on the descending slope of the PVER amplitude-spatial frequency function toward the higher spatial frequency to the baseline. RESULTS: The PL acuities ranged from 20/25 to < 20/1600 (mean 20/155). The correlation between the two methods was good (r = 0.847). Fifty-six patients (70%) had an acuity agreement within 1.0 octave. When the PL acuity was > 20/128, it was on average better than the PVER acuity. When the PL acuity was lower, the PVER acuity was usually better. This tendency was marked when the visual acuities were very poor (y = 0.552x + 0.362). CONCLUSION: The methods correlate well, although there is a dissociation of acuities in the presence of very low vision. PVER may be a useful addition to PL in assessment of vision in infants and young children. PMID- 9407227 TI - Toward robot-assisted vascular microsurgery in the retina. AB - BACKGROUND: Experimental protocol in our laboratory routinely requires the precise placement of instruments at, or near, the retina. Although manipulators for placing an instrument within the eye presently exist, none of the designs were satisfactory due to limitations on size, accuracy and operability. To overcome these limitations, we have developed a novel six degree of freedom manipulator designed specifically for retinal microsurgery. METHODS: The manipulator is parallel in structure and provides submicrometer positioning of an instrument within the constrained environment of the eye. The position of an instrument attached to the manipulator is commanded by the operator using a hand held trackball. A computer controller interprets the trackball input and moves the manipulator in an intuitive manner according to mathematically constrained modes of operation. RESULTS: Over 50 retinal vessels in the live, anesthetized cat have been successfully cannulated for pressure measurement and drug injection using the described manipulator and micropuncture techniques. The targeted vessels ranged in internal diameter from 20 to 130 microns. CONCLUSION: This device has applications in microsurgery where tremor and fatigue limit the performance of an unaided hand and where mechanically constrained manipulators are inappropriate due to size and operative constraints. PMID- 9407228 TI - Hyperopia correction by noncontact holmium: YAG laser thermal keratoplasty: five pulse treatments with 1-year follow-up. AB - BACKGROUND: Previous noncontact holmium (Ho): YAG laser thermal keratoplasty (LTK) studies on correction of low to moderate hyperopia have used treatment algorithms based on ten-pulse, variable-pulse-energy treatment parameters. The purpose of this study was to evaluate the safety, effectiveness, and stability of new five-pulse, constant-pulse-energy treatment parameters for noncontact Ho:YAG LTK. METHODS: Thirty-nine hyperopic patient eyes [up to +4.75 diopters (D) refractive error] were treated using simultaneous noncontact delivery of Ho:YAG laser energy (Sunrise) with two symmetrical octagonal rings of eight spots per ring and radial spot patterns on centerline diameters of 5 and 6 mm (group A), 6 and 7 mm (group B), or 6.5 and 7.5 mm (group C). Each ring of spots received five pulses of laser light at 5 Hz pulse repetition frequency and a fixed pulse energy of 240 mJ. Thirty of the 39 patient eyes (77%) had 1-year follow-up exams. RESULTS: At 1 year, the mean Snellen uncorrected distance visual acuity lines gained was 3.7 +/- 0.5/6.8 +/- 2.7/5.3 +/- 3.3 for groups A, B, and C. The mean changes in subjective manifest refraction (spherical equivalent) were -2.08 +/- 1.13 D, -1.83 +/- 0.88 D, -1.22 +/- 0.88 D for groups A, B, and C respectively. None of the eyes lost two or more lines of spectacle-corrected distance visual acuity. There were no clinically significant complications in any patient. CONCLUSION: This clinical study indicates that five-pulse noncontact LTK treatments of low hyperopia are safe and effective. The stability has to be confirmed with longer follow-up. PMID- 9407229 TI - Influence of viral infection on expression of cell surface antigens in human retinal pigment epithelial cells. AB - BACKGROUND: Subacute viral infection is known to change the phenotype of infected cells, thereby causing immune-mediated tissue damage. The aim of this study was to investigate the expression of different cell surface molecules on human retinal pigment epithelial cells (RPEC) following viral infection, with special emphasis on those having immune-regulatory functions. METHODS: Cultured RPEC were infected with cytomegalovirus (CMV), coxsackie-virus B3 (CVB) or herpes simplex virus type I (HSV). Double-staining fluorescence technique was used for visualization of virus infection and cell surface markers in the same cells by laser microscopy. RESULTS: CMV downregulated MHC class I antigens on RPEC, whereas CVB and HSV did not alter MHC class I antigen expression. No induction of class II antigens was observed in RPEC infected with CVB, HSV or CMV. The intercellular adhesion molecule ICAM-1 (CD54) was strongly expressed in uninfected RPEC, and a slight increase was observed after virus infection. Vascular cell adhesion molecule 1 (VCAM-1) was expressed in low amounts in both uninfected and infected RPEC. No expression of intercellular adhesion molecule 2 (ICAM-2), E-selectin ELAM-1 or lymphocyte-function-associated antigen 1 (LFA-1) was observed on RPEC before or after virus infection. CONCLUSION: Downmodulation of immune-regulating cell surface antigens has been suggested to provide a means of long-term survival of viruses in the infected cell, favoring establishment of persistent infection. Our observation in cultured human RPEC indicates that this mechanism might indeed contribute to the development of disease affecting retinal tissue. PMID- 9407230 TI - Morphological differentiation of the conjunctival goblet cells in the chick (Gallus domesticus). AB - BACKGROUND: These is no consensus in the literature regarding the differentiation of conjunctival goblet cells in vertebrates. METHOD: The conjunctival epithelium of the chick was studied before and after hatching in order to demonstrate the morphological evolution of the goblet cells. The entire conjunctiva was processed for light microscopy either on semithin sections stained with toluidine blue pironine or on traditional sections stained with Alcian blue pH 2.5-PAS. RESULTS: It was possible to demonstrate that goblet cells underwent remarkable changes in their secretory activity. At 12 h after hatching, isolated Alcian blue-positive cells were present in the fornix. At 24 h after hatching, cells positive for both Alcian blue and PAS were scattered among epithelial cells. Two days after hatching, cells which reacted positively only to PAS were also present. CONCLUSION: It is suggested that the differentiation of conjunctival goblet cells occurs first in the fornix, probably due to the particular vascular environment of this region, and then spreads all over the conjunctiva. PMID- 9407231 TI - The effect of high-dose methylprednisolone on laser-induced retinal injury in primates: an electron microscopic study. AB - BACKGROUND: Previously we reported an ameliorative effect of high-dose methylprednisolone in laser injury to monkey retinas. The ultrastructural modification by methyl-prednisolone has not been examined. METHODS: Cynomolgus monkeys were given severe (grade III) retinal laser burns and treated with an intravenous megadose of methylprednisolone. Pathologic features of the retinal lesions with or without methylprednisolone treatment were evaluated by light and electron microscopy. RESULTS: Ultrastructurally, the treated lesions showed rapid recanalization of choriocapillaris; proliferation of retinal pigment epithelium to replace the necrotic and damaged cells, resulting in rapid re-establishment of blood retinal barrier; mild macrophagic activity; and rapid reformation of the outer limiting membrane by Mueller cells. CONCLUSION: A high dose of methylprednisolone affected the responses of the choriocapillaris, retinal pigment epithelium, photoreceptor cells and Mueller cells to laser injury, showing an overall beneficial effect. These modifications might be ascribed to methylprednisolone's anti-inflammatory action, protection of the microcirculation and anti-lipid peroxidation effect. PMID- 9407233 TI - Preoperative vitreous hemorrhage associated with rhegmatogenous retinal detachment. PMID- 9407234 TI - Triage decisions for intensive care in terminally ill patients. PMID- 9407232 TI - Fibroblast growth factor 2, heparin and suramin reduce epithelial ulcer development in experimental HSV-1 keratitis. AB - BACKGROUND: We have previously shown that basic fibroblast growth factor (FGF-2) enhances corneal epithelial healing in different experimental models in vivo. In order to study the healing effect of this growth factor in pathological conditions of the cornea, we investigated whether topical application of FGF-2 could affect herpes keratitis in rabbits. Since HSV-1 infection is prevented in vitro by incubation with heparin, we also topically applied heparin and suramin, considering the similar interaction of herpes simplex virus and FGF-2 with cell membrane-anchored heparan sulfate. METHODS: After virus inoculation with a human BEY.2 strain, rabbits were treated with either FGF-2 (200 ng to 2 micrograms/application), heparin (250 micrograms/application) or suramin (250 micrograms/application) 4 times daily until day 14. Acyclovir and placebo administrations served as controls (n = 48 rabbits). Computerized ulcer surface analysis, clinical observations and virus recovery assays were performed. RESULTS: Topical FGF-2, heparin and suramin treatment revealed a significant reduction in peak ulcer sizes, and complete epithelial healing was achieved earlier than in placebo-treated corneas. However, no significant antiviral effect of FGF-2, heparin and suramin was detectable in plaque assays from conjunctival swabs. CONCLUSIONS: These experiments demonstrate that FGF-2 is effective in promoting herpetic epithelial ulcer healing, either due to its proliferative effects on epithelial cells or indirectly by occupying the sites on cell surface heparan sulfate necessary for the attachment of the virion. The latter mechanism of action is presumably the reason for the similar effect of heparin and suramin. PMID- 9407235 TI - Hypothermia: an adverse effect or a missing partner? PMID- 9407236 TI - Septic shock in patients with the acquired immunodeficiency syndrome. AB - OBJECTIVE: To evaluate the prognosis of patients with septic shock admitted to an intensive care unit (ICU), according to their HIV serostatus. DESIGN: Retrospective study. SETTING: Medical ICU of a university hospital. PATIENTS: 76 patients with septic shock admitted to the same ICU, of whom 28 were HIV positive and 48 were HIV negative. MEASUREMENTS AND RESULTS: Severity scores, number and type of organ failures, and survival rates were assessed in the two groups of patients. Glasgow Coma Scale and general severity scores [Acute Physiology and Chronic Health Evaluation II and Simplified Acute Physiology Score (SAPS)] were significantly worse in HIV-infected patients. The total number of organ failures was also higher in the HIV-positive group: 3.7 +/- 0.2 vs 3.1 +/- 0.2 in the HIV negative group (p < 0.001). On day 28, 21 (46%) HIV-negative patients were dead compared to 26 (93%) patients in the HIV-positive group (p < 0.001). In the multivariate analysis, HIV infection was an independent risk factor for mortality, as were the SAPS score, use of mechanical ventilation, and the McCabe score. CONCLUSIONS: This study reports a considerable excess mortality in HIV infected patients with septic shock. Although severity of illness was clearly much more pronounced in HIV-positive patients, retroviral infection was independently associated with death. Improving survival in HIV-positive patients with septic shock may require earlier diagnosis and treatment of the causative infection. PMID- 9407237 TI - Facial mask noninvasive mechanical ventilation reduces the incidence of nosocomial pneumonia. A prospective epidemiological survey from a single ICU. AB - OBJECTIVE: To evaluate the impact of noninvasive positive pressure mechanical ventilation (NPPV) on ventilator-associated pneumonia (VAP). DESIGN: Prospective observational study. SETTING: Medical intensive care unit (ICU) of a university teaching hospital. PATIENTS: Cohort of 320 consecutive patients staying in the ICU more than 2 days and mechanically ventilated for > or = 1 day. MEASUREMENTS AND RESULTS: VAP was diagnosed when, satisfying classical clinical and radiological criteria, fiberoptic bronchoalveolar lavage and/or protected specimen brush grew > or = 10(4) and > or = 10(3) CFU/ml, respectively, of at least one microorganism. Patients were classified into four subgroups according to the way in which mechanical ventilation was delivered: NPPV then tracheal intubation (TI) (n = 38), TI then NPPV (n = 23), TI only (n = 199), and NPPV only (n = 60). Occurrence of VAP was estimated by incidence rate and density of incidence. Risk factors for VAP were assessed by logistic regression analysis. Twenty-seven patients had 28 episodes of VAP. The incidence rates for patients with VAP were 18% in NPPV-TI, 22% in TI-NPPV, 8% in TI, and 0% in NPPV (p < 0.0001). The density of incidence of VAP was 0.85 per 100 days of TI and 0.16 per 100 days of NPPV (p = 0.04). Logistic regression showed that length of ICU stay and ventilatory support were associated with VAP. CONCLUSIONS: There is a significantly lower incidence of VAP associated with NPPV compared to tracheal intubation. This is mainly explained by differences in patient severity and risk exposure. PMID- 9407238 TI - Short-term effects of prone position in critically ill patients with acute respiratory distress syndrome. AB - OBJECTIVE: Changing the position from supine to prone is an emerging strategy to improve gas exchange in patients with the acute respiratory distress syndrome (ARDS). The aim of this study was to evaluate the acute effects on gas exchange, hemodynamics, and respiratory system mechanics of turning critically ill patients with ARDS from supine to prone. DESIGN: Open, prospective study. SETTING: General intensive care units. PATIENTS: 23 patients [mean age 56 +/- 17 (SD) years] who met ARDS criteria and had a Lung Injury Score > 2.5 (mean 3.25 +/- 0.3). INTERVENTIONS: The decision to turn a patient was made using a protocol based on impaired oxygenation despite the use of positive end-expiratory pressure and a fractional inspired oxygen (FIO2) of 1. MEASUREMENTS AND RESULTS: We measured gas exchange and hemodynamic variables in all patients and in 16 patients calculated respiratory system compliance when they were supine and 60 to 90 min after turning them to a prone position. This latter position was remarkably well tolerated and no clinically relevant complications or events were detected either during turning or while prone. The partial pressure of oxygen in arterial blood (PaO2)/FIO2 ratio improved from 78 +/- 37 mm Hg supine to 115 +/- 31 mm Hg prone (p < 0.001), and intrapulmonary shunt decreased from 43 +/- 11 to 34 +/- 8% (p < 0.001). Cardiac output and other hemodynamic parameters were not affected. Respiratory system compliance slightly improved from 24.7 +/- 10.2 ml/cmH20 supine to 27.8 +/- 13.2 ml/cmH20 prone (p < 0.05). An improvement in PaO2/FIO2 of more than 15% from changing from supine to prone was found in 16 patients (responders). Responders had more hypoxemia (PaO2/FIO2 70 +/- 23 vs 99 +/- 53 mm Hg in non-responders, p < 0.01), more hypercapnia (partial pressure of carbon dioxide in arterial blood (70 +/- 27 vs 64 +/- 9 mm Hg, p < 0.01) and a shorter elapsed time to the onset of ARDS and turning to the prone position (11.8 +/- 16 vs 32.8 +/- 42 days, p < 0.01). CONCLUSIONS: Turning critically ill, severely hypoxemic patients from the supine to the prone position is a safe and useful therapeutic intervention. Our data suggest that prone positioning should be carried out early in the course of ARDS. PMID- 9407240 TI - Comparison of electrocardiograms recorded with standard leads and derived from the vectorcardiographic frank leads in high risk patients. AB - Dynamic vectorcardiography (VCG) is increasingly employed for ischaemia monitoring with the use of a computerized method for recording and on-line analysis by the calculation of trend parameters. To elucidate how well the derived electrocardiogram (dECG), calculated from the VCG, compares with the simultaneously registered standard ECG (sECG), dECGs from 17 postoperative cardiac-risk patients and 36 subjects with acute myocardial infarction (AMI) were compared to sECGs, both quantitatively in leads II, III, V2 and V5 and qualitatively. Despite small, but some significant differences, mainly in the amplitudes of precordial leads, the qualitative interpretation by two independent cardiologists showed good agreement between the methods (kappa = 0.72 and 0.67, respectively) for the diagnosis of AMI/ischaemia. The dECG seems to be reliable and can be used clinically in these groups of patients during VCG recordings. PMID- 9407239 TI - Is endotoxin and cytokine release related to a decrease in gastric intramucosal pH after hemorrhagic shock? AB - OBJECTIVES: (a) To investigate the relationship between gut ischemia parameters (gastric intramucosal pH [pHi], mucosal-arterial carbon dioxide difference [PCO2 gap]), and endotoxin or cytokine release during hemorrhagic shock; (b) to compare the predictive value of pHi, PCO2-gap and arterial lactate concentrations. DESIGN: Prospective study. SETTING: Surgical intensive care unit of a university hospital. PATIENTS: 20 multiple trauma patients with severe hemorrhagic shock. INTERVENTIONS: Intramucosal measurements and blood samples were obtained on admission to the emergency room and repeatedly over 48 h. MEASUREMENTS AND RESULTS: Endotoxin was measured using a chromogenic limulus amoebocyte assay. Cytokine [tumor necrosis factor-alpha (TNF alpha) and interleukin-6 (IL-6)] values were evaluated by immunoradiometric assays. Only 3 patients had positive blood cultures but endotoxins were detected at least once in all patients. Endotoxin levels were similar in survivors and non-survivors over the study period and were not related to pHi or PCO2-gap. Initially, high levels of IL-6 were observed in both nonsurvivors and survivors [median 1778 pg/ml (range 435 44,540) vs 2068 pg/ml (range 996-92,300)]. IL-6 levels progressively decreased in the survivors but not significantly. On admission, TNF alpha concentrations were similar in nonsurvivors and survivors (42 +/- 35 vs 46 +/- 27 pg/ml). From the 24th h, TNF alpha values were higher in the nonsurvivors than in the survivors (24 h: 72 +/- 38 vs 34 +/- 17 pg/ml, p < 0.05). The greatest IL-6 levels were found for a pHi < 7.20 (28.5 +/- 36.5 vs 1.8 +/- 1.3 ng/ml, p < 0.05) or a PCO2 gap > 7.5 mmHg (1 kPa) (32.5 +/- 37.5 vs 1.7 +/- 1.3 ng/ml, p < 0.01). With the same pHi threshold, no difference was found in endotoxin levels. The lactate concentrations were predictive for outcome from the 12th h (9.5 +/- 5.9 vs 3.6 +/ 2.3 mmol/l, p < 0.05). CONCLUSIONS: During severe hemorrhagic shock, endotoxin translocation from the gut was a common phenomenon that seemed independent of both pHi values and outcome. It could not explain IL-6 and TNF alpha release. In severe hemorrhagic shock, neither pHi nor PCO2-gap provides additional information to the lactate measurements. PMID- 9407241 TI - The ability of the Simplified Acute Physiology Score (SAPS II) to predict outcome in coronary care patients. AB - OBJECTIVE: To evaluate the applicability of the Simplified Acute Physiology Score (SAPS II) for coronary care patients. DESIGN: Prospective observational cohort study. SETTING: Medical ICU of a community teaching hospital. PATIENTS: 1587 consecutive patients admitted over a period of 18 months. MEASUREMENTS AND MAIN RESULTS: Patients were divided in two groups according to the primary admission diagnosis: general medical intensive care (ICU) patients and intensive coronary care (CCU) patients. Score prediction was tested using criteria suitable to evaluate the discrimination and calibration properties of SAPS II. Mean SAPS II score was 31.6 (+/- 20.1) in ICU and 28.3 (+/- 15.5) in CCU patients (p = 0.06), mean risk of death 0.206 and 0.134 (p = 0.001), and observed hospital mortality 17.8 vs 10.3%. The area under the receiver operating characteristic curve was 0.888 in ICU and 0.908 in CCU patients (p = 0.5). The correlation between predicted and observed hospital mortality was 0.62 (p = 0.001) in ICU and 0.66 (p = 0.001) in CCU patients. The calibration curves did not differ from each other. The probability of death in survivors and nonsurvivors was equally distributed in ICU and CCU patients (p = 0.5). CONCLUSION: We conclude that SAPS II is applicable to CCU patients in our unit. PMID- 9407242 TI - Congenital diaphragmatic hernia: antenatal prognostic factors. Does cardiac ventricular disproportion in utero predict outcome and pulmonary hypoplasia? AB - Despite regular progress in neonatal intensive care, congenital diaphragmatic hernia (CDH) diagnosed antenatally is still associated with up to 80% mortality. It is impossible to predict which fetus with CDH will survive or not. OBJECTIVE: To identify reliable antenatal predictors of outcome and of pulmonary hypoplasia (PH) in fetuses with CDH. DESIGN: Retrospective study. SETTING: Paediatric intensive care unit of a university children's hospital. PATIENTS AND METHODS: Antenatal parameters and presence of left ventricular hypoplasia in utero were compared retrospectively to outcome and to presence of PH in 32 consecutive newborn infants with antenatally diagnosed CDH. Antenatal parameters included: gestational age at diagnosis, herniated organs, associated malformations and presence of polyhydramnios. Size of the cardiac ventricles, the aorta (Ao) and the pulmonary artery (PA) were obtained by fetal echocardiography, from which we calculated a cardioventricular index (left ventricle/right ventricle, LV/RV) and a cardiovascular index (Ao/PA). Delivery was planned in order to provide ventilatory and hemodynamic management. In case of death, PH was assessed according to the following criteria: the lung weight/body weight index and the radial alveolar count. For statistical comparisons, patients were separated into two groups: the hypoplasia group (H) and the non-hypoplasia group (NH). RESULTS: Thirty-two pregnancies were delivered. Twenty-six newborns died (81%), 6 survived (19%). When comparing non-survivors to survivors, predictors of poor outcome were: mean gestational age at diagnosis (23 vs 28 weeks, p = 0.002), intrathoracic stomach (20 vs 1 s, p = 0.01) and associated malformations (6 vs 0). Cardiac ventricular disproportion, expressed by the LV/RV ratio, appeared to correlate well with a poor outcome (0.63 in non-survivors vs 0.93 in survivors, p = 0.03) and with PH (0.63 in the H group vs 0.95 in the NH group, p = 0.03). CONCLUSIONS: Our study confirmed the factors for a poor prognosis associated with CDH previously described in the literature, but none with a consistent demonstration of accuracy. LV hypoplasia may be a more accurate predictor of outcome and of PH but it has to be assessed by prospective studies with larger samples. Further basic science and Doppler-flow studies may be helpful to understand the natural history and pathophysiology of LV hypoplasia in CDH. PMID- 9407243 TI - Surfactant nebulisation: lung function, surfactant distribution and pulmonary blood flow distribution in lung lavaged rabbits. AB - OBJECTIVE: Surfactant nebulisation is a promising alternative to surfactant instillation in newborns with the respiratory distress syndrome. Although less surfactant is deposited in the lung, it improves gas exchange, probably due to a superior distribution. We hypothesize that a more uniform distribution of nebulised surfactant results in a more uniform pulmonary blood flow and consequently a more efficient gas exchange. We asked whether the pulmonary blood flow changes after surfactant replacement, and to what extent pulmonary blood flow is influenced by the amount of surfactant deposition. Furthermore, we investigated whether sufficient nebulised surfactant is deposited in the lungs to achieve a sustained improvement in lung function. INTERVENTIONS: Surfactant was nebulised or instilled, or saline was nebulised, in 18 lung-lavaged rabbits. After 2 h the rabbits were weaned from mechanical ventilation to continuous positive airway pressure, 40% oxygen. We measured blood gasses, dynamic lung compliance, surfactant distribution using 99m technetium nanocoll label, and the pulmonary blood flow distribution, using microspheres. RESULTS: Partial pressure of oxygen in arterial blood and lung compliance were significantly higher after surfactant nebulisation than after saline nebulisation. Surfactant instillation gave a superior effect with respect to these variables. Nebulised surfactant was distributed more uniformly over the lungs than instilled surfactant. Although pulmonary blood flow changed over time, it remained uniformly distributed following both modes of surfactant treatment. Surfactant deposition was neither strongly related to pulmonary blood flow nor strongly related to the change in blood flow. CONCLUSIONS: Although nebulised surfactant is uniformly distributed, we can provide no evidence that this results in a more uniform pulmonary blood flow distribution. Therefore, other than a superior surfactant distribution, no additional reason was found for the efficient gas exchange after nebulisation. PMID- 9407244 TI - Surfactant nebulisation prevents the adverse effects of surfactant therapy on blood pressure and cerebral blood flow in rabbits with severe respiratory failure. AB - OBJECTIVE: Surfactant replacement therapy for the neonatal respiratory distress syndrome has shown beneficial effects on lung function and survival. Recently, rapid fluctuations of haemodynamics and cerebral perfusion following surfactant instillation have been described and an association with the development of intraventricular haemorrhage has been proposed. Therefore, alternative methods of surfactant therapy that reduce the effects on cerebral perfusion have to be explored. Does instillation of surfactant influence blood pressure and cerebral blood flow in rabbits with severe respiratory failure? Can nebulisation of surfactant prevent these adverse effects on blood pressure and cerebral blood flow? INTERVENTIONS: Surfactant (Alveofact, 100 mg/kg body weight) was nebulised using the MiniNEB nebuliser, or instilled, in 12 rabbits with severe respiratory failure induced by lung lavage. Assessed were blood gasses, mean arterial blood pressure (MABP) and cerebral blood flow over the left carotid artery, using ultrasonic transit-time flow probes. RESULTS: Partial pressure of oxygen in arterial blood increased quickly after instillation, from 8.7 +/- 1.3 to 24.9 +/- 6.4 kPa after 15 min, and increased gradually during nebulisation from 8.0 +/- 0.5 to 24.5 +/- 4.6 after 120 min. After instillation, MABP decreased 22 +/- 5% (in 8 min) and cerebral blood flow dropped even more: 64 +/- 9% within 8 min. During nebulisation, MABP did not change significantly and cerebral blood flow decreased gradually, 31 +/- 14% over 90 min. CONCLUSIONS: Surfactant instillation was followed by a rapid decrease in MABP and an even more pronounced drop in cerebral blood flow, while during nebulisation MABP did not change and cerebral blood flow decreased less and more gradually. PMID- 9407245 TI - Interaction between haematocrit and pulmonary blood volume on pulmonary vascular flow resistance and pressure flow relationships. AB - OBJECTIVE: Pulmonary vascular flow resistance depends on blood viscosity, mainly due to haematocrit, and on vessel dimensions determining blood volume in this highly compliant vascular bed. We, therefore, evaluated the interaction between haematocrit, blood flow, and transpulmonary vascular pressure gradient under conditions of controlled pulmonary blood volume. DESIGN: Experimental study in isolated zone-III rabbit lungs perfused with autologous blood. SETTING: Laboratory for experimental studies. INTERVENTIONS: Stepwise and independent variation of flow (50, 100, and 200 ml/min), pulmonary blood volume (increments of 2.5 ml and 5 ml imposed by changes of left atrial pressure), and haematocrit (0-50%) varied by haemodilution (Krebs-Henseleit/albumin) or haemoconcentration (centrifugation). MEASUREMENTS: Pulmonary arterial, left atrial, and airway pressures as well as reservoir volume (reflecting reciprocal changes of lung blood volume) and lung weight. RESULTS: Haemodilution from the normal haematocrit (32%) to 10% at constant pulmonary blood volume and flow decreased flow resistance only slightly, whereas haemoconcentration (50%) increased flow resistance up to 130%. At the same time increments of in pulmonary blood volume of 2.5 and 5 ml (approx. 15 and 30% of normal pulmonary blood volume) at constant haematocrit significantly shifted downwards pressure-flow relationships for all investigated haematocrits (0-50%). CONCLUSIONS: Because of the multiple interrelationships between haematocrit, blood flow and pulmonary blood volume, haematocrit effects on pulmonary flow resistance and pressure-flow relationships in the pulmonary vasculature should be studied at controlled blood volume. While haemodilution only has minor effects, haemoconcentration changes pressure-flow relationships markedly. Pulmonary blood volume has a major impact on slope and position of pressure-flow relationships for all haematocrits investigated. PMID- 9407247 TI - Communication in the ICU. PMID- 9407246 TI - Inhaled nitric oxide in patients with pulmonary embolism. AB - OBJECTIVE: To describe the use of inhaled nitric oxide (NO) in four patients with severe pulmonary embolism. SETTING: The intensive care unit (ICU) of a university teaching hospital. PATIENTS: Four patients with severe pulmonary embolism on the basis of clinical, haemodynamic or blood-gas parameters received NO by inhalation either during spontaneous respiration (two cases) or while mechanically ventilated (two cases). INTERVENTIONS: Conventional management of pulmonary embolism in addition to the use of inhaled NO. MEASUREMENTS AND RESULTS: Description of clinical course, haemodynamic and gas-exchange data. Dose-response data are also described for three patients. CONCLUSIONS: We reported four cases of pulmonary embolism where the administration of inhaled NO resulted in an improvement in pulmonary haemodynamic and gas-exchange parameters. Two patients were weaned from NO and survived until discharged from the ICU. Inhaled NO might be a useful adjunct in pulmonary embolism to improve stability of the patient prior to thrombolysis or surgery. PMID- 9407248 TI - Fatal hepatic toxicity of DTIC: a new case. PMID- 9407250 TI - The hemodynamic consequences of mechanical ventilation. PMID- 9407249 TI - Factitial peritonitis after radiotherapy for locally advanced cancer of the cervix. PMID- 9407251 TI - Gastrointestinal permeability in cardiac surgery patients, the interpretation of dual-sugar absorption studies. PMID- 9407253 TI - Carbohydrate drinks delay fatigue during intermittent, high-intensity cycling in active men and women. AB - The effects of ingesting carbohydrate drinks on fatigue during intermittent, high intensity cycling in men and women were determined. Physically active but untrained women (n = 7) and men (n = 9) completed one practice trial and two experimental sessions separated by 1 week. Sessions consisted of repeated 1-min cycling bouts on a bicycle ergometer at 120-130% VO2max separated by 3 min rest until fatigue. Carbohydrate (CHO) or placebo (P) beverages (4 ml.kg body weight 1) were ingested immediately before exercise (18% CHO) and every 20 min during exercise (6% CHO). Plasma glucose and insulin were higher, RPE for the legs was lower, and time to fatigue was longer in CHO than P. Men's and women's responses were not different for any variable measured. These data suggest a beneficial role of CHO drinks on performance of intermittent, high-intensity exercise in men and women. PMID- 9407252 TI - Carbohydrate intake and recovery of intermittent running capacity. AB - The purpose of the present study was to examine the influence of an increased carbohydrate intake on the recovery of endurance running capacity after exhaustive intermittent running. Six male subjects were randomly assigned to two dietary recovery conditions, each involving two running tests separated by 22 hr. The protocol comprised a prolonged, intermittent, high-intensity shuttle run test (I-HI). One week later subjects repeated the I-HI on consecutive days under different dietary conditions. During the 22-hr recovery, either the carbohydrate intake of the subjects was increased (CHO) or they ate an isocaloric diet by supplementing their normal diet with extra protein and fat (CON). Intermittent running capacity was improved when subjects increased their carbohydrate intake to 10 g.kg-1 bm during the 22-hr recovery between trials, but an isocaloric diet without additional carbohydrate did not bring about the same improvements. PMID- 9407254 TI - Effect of carbohydrate substrate availability on ratings of perceived exertion during prolonged running. AB - The purpose of this study was to investigate the effects of carbohydrate substrate availability on ratings of perceived exertion (RPE) during prolonged submaximal running. Thirty marathon runners were recruited as subjects. A double blind study design was used in which subjects performed an experimental trial that consisted of a 2.5-hr treadmill run at 75-80% VO2max. During the experimental trial, the subjects in the carbohydrate feeding group ingested a 6% glucose and fructose solution at a rate of approximately 60 g.hr, whereas subjects in the placebo group consumed an equal volume of artificially flavored placebo. Statistical analysis of RPE, respiratory exchange ratio, fat and carbohydrate oxidation rate, and blood glucose concentrations indicated that increased carbohydrate substrate availability attenuated the intensity of exertional perceptions during the later stages of prolonged running at 75-80% VO2max in marathon runners. PMID- 9407255 TI - The effects of beverage carbonation on sensory responses and voluntary fluid intake following exercise. AB - The effects of carbonated beverages on sensory acceptability and voluntary fluid intake after exercise were examined. The level of carbonation in a 6% carbohydrate (CHO) electrolyte drink was systematically varied (0, 1.1, 2.3, and 3.0 volumes of CO2), and its impact was assessed in 52 adults following 30 min of exercise. The perception of carbonation intensity closely tracked the differences in physical carbonation levels presented, with all perceived intensities significantly different from each other (p < .01). Overall sensory acceptability, perceived thirst quenching, and perceived sweetness were significantly lower for 2.3-vol CO2 and 3.0-vol CO2 than for 0-vol CO2 and 1.1-vol CO2 (p < .01). Perceived throatburn was significantly higher for 2.3-vol CO2 and 3.0-vol CO2 than for 0-vol CO2 and 1.1-vol CO2 (p < .01). Total fluid intake for 0-vol CO2 and 1.1-vol CO2 was significantly higher than for 2.3-vol CO2 (p < .05), which was significantly higher than for 3.0-vol CO2 (p < .05). It was concluded that levels of carbonation equal to or in excess of 2.3-vol CO2 negatively impact drink acceptability and voluntary fluid intake. PMID- 9407256 TI - Effects of a low-dose amino acid supplement on adaptations to cycling training in untrained individuals. AB - The purpose of this study was to determine if amino acid supplementation influences blood and muscle lactate response to exercise and the time course of the metabolic adaptations to training. Two groups of untrained males (n = 7 each) were given (double-blind) a daily supplement (2.9 g.day-1) containing a mixture of leucine, isoleucine, valine, glutamine, and carnitine (EXP) or 3 g.day-1 of lactose (CON). Following 7 days of supplementation there was no significant change in VO2peak, time to exhaustion (TTX) at 120% VO2peak, or muscle and blood lactate in either EXP or CON. Subjects then initiated 6 weeks of combined aerobic and anaerobic training on a Monark cycle ergometer. It was found that amino acid supplementation had no effect on either blood or muscle lactate accumulation during exercise, while supplementation resulted in a faster adaptation in buffer capacity. Performance during intense exercise was not improved with amino acid supplementation. PMID- 9407257 TI - The role of social physique anxiety and other variables in predicting eating behaviors in college students. AB - Early identification of potentially harmful eating patterns is critical in the effective remediation of such behaviors. The purpose of this investigation was to examine the degree to which various factors including gender, family history, and athletic status predict disordered eating behavior; social physique anxiety and percent body fat were added as potential predictor variables. The eating behaviors of student-athletes and nonathlete students were also compared. One hundred eighty undergraduate students (males = 49, females = 131) provided demographic information and completed the Eating Attitudes Test (EAT) and the Social Physique Anxiety Scale (SPAS). Stepwise multiple-regression analysis indicated that social physique anxiety, gender, and body fat (%Fat) combined to predict 34% of disordered eating behaviors: EAT = 0.921 SPA - 1.05 %Fat + 10.95 Gender (1 = M, 2 = F) - 17.82 (R2 = .34, SE = 4.68). A one-way ANOVA comparing the eating behaviors of athletes and nonathletes revealed no significant difference between these groups. PMID- 9407258 TI - The effects of gamma-oryzanol supplementation during resistance exercise training. AB - To determine the effectiveness of gamma-oryzanol supplementation, weight-trained males were randomly divided into supplemented (G-O) and control placebo (Con) groups. The G-O group ingested 500 mg.day-1 of gamma-oryzanol according to manufacturer's instructions. Test batteries were administered before (T1), after 4 weeks (T2), and after 9 weeks (T3) of a periodized resistance exercise program. Both groups demonstrated significant increases in 1 repetition maximum muscular strength (bench press and squat) and vertical jump power, with no differences between the groups. No differences between groups were observed for measures of circulating concentrations of hormones (testosterone, cortisol, estradiol, growth hormone, insulin, beta-endorphin), minerals (calcium, magnesium), binding protein (albumin), or blood lipids (total cholesterol, triglycerides, HDL-cholesterol). Resting cardiovascular variables decreased similarly for both groups. These data suggest that 9 weeks of 500 mg.day-1 of gamma-oryzanol supplementation does not influence performance or related physiological parameters in moderately weight trained males. PMID- 9407260 TI - Effects of a nutritional intervention on triathletes' energy intakes. PMID- 9407259 TI - Effects of creatine supplementation on repetitive sprint performance and body composition in competitive swimmers. AB - In a double-blind and randomized manner, 18 male and female junior competitive swimmers supplemented their diets with 21 g.day-1 of creatine monohydrate (Cr) or a maltodextrin placebo (P) for 9 days during training. Prior to and following supplementation, subjects performed three 100-m freestyle sprint swims (long course) with 60 s rest/recovery between heats. In addition, subjects performed three 20-s arm ergometer maximal-effort sprint tests in the prone position with 60 s rest/recovery between sprint tests. Significant differences were observed among swim times, with Cr subjects swimming significantly faster than P subjects following supplementation in Heat 1 and significantly decreasing swim time in the second 100-m sprint. There was also some evidence that cumulative time to perform the three 100-m swims was decreased in the Cr group. Results indicate that 9 days of Cr supplementation during swim training may provide some ergogenic value to competitive junior swimmers during repetitive sprint performance. PMID- 9407261 TI - Medical accidents in hospital care: applications of failure analysis to hospital quality appraisal. AB - BACKGROUND: Medical accidents can be understood as patient injuries that result from interaction of physician or nurse error during the provision of care with faults latent in the hospital system. Medical accidents are not random events but are events with discoverable associations between human error and system faults through application of methods of failure analysis in the evaluation of patient injuries. CASE ANALYSIS: The goal of a failure analysis is to make apparent system faults that are otherwise obscured. Analyses seek to answer several questions. What characteristics of the system failed to prevent a slip, mistake, or rule violation from evolving into an accident? What system changes might have offset, or prevented, the active error from contributing to the sequence of events culminating in injury? Brief descriptions of eight cases of apparent medical accidents are provided in this article. For three of these cases, the failure analysis approach is used to identify the sequence of events contributing to the patient injury; identify events within this sequence that represent active errors; and identify points within this sequence that represent system faults which failed to prevent the occurrence of subsequent events. CONCLUSIONS: Within the framework of current methods of hospital quality appraisal, attribution of patient injury historically has focused on clinician error. Yet unless detected and corrected, system faults persist and create circumstances of "accidents waiting to happen." Understanding of casual factors in the evolution of medical accidents can be usefully applied toward improvement in the quality of hospital appraisal of iatrogenic injuries and, through that application, toward reduction in the rates of adverse outcomes. PMID- 9407262 TI - Using continuous quality improvement to improve diabetes care in populations: the IDEAL model. Improving care for Diabetics through Empowerment Active collaboration and Leadership. AB - BACKGROUND: The care of patients with chronic diseases, especially those with diabetes mellitus, has been less than ideal. However, despite clear national guidelines, various examples of better care models, and multiple attempts to improve care, an effective process for facilitating and replicating diabetes care improvements in typical primary care practices has been elusive. METHODS: On the basis of the approach and lessons from developmental work at the Minnesota Diabetes Control Program and a trial of continuous quality improvement for clinical preventive services (IMPROVE), a clinic-based intervention processes (IDEAL) has been developed to improve the system and process of care for patients with diabetes as a model for all chronic diseases. The intervention incorporates facilitation of leadership actions in support of change, training for the leader and facilitator of an intraclinic multidisciplinary continuous quality improvement (CQI) team, and consultative and networking support of the change process. Each element of this intervention emphasizes a seven-step process improvement approach and a system for care of patients with diabetes. This model is being developed and tested in a unique partnership between the Minnesota Department of Health and HealthPartners, a large managed care organization (MCO). RESULTS: A prepilot demonstration has succeeded in improving glycemic control, three primary care clinics affiliated with HealthPartners have succeeded in a pilot of the intervention, and an additional 13 clinics are participating in a randomized controlled trial of a refined intervention. CONCLUSIONS: The IDEAL model holds promise for substantial improvements in care, not only for diabetes but for all chronic diseases and for other settings. PMID- 9407263 TI - Implementing programs for chronic illness management: the case of hypertension services. AB - BACKGROUND: This article describes the process by which HealthSystem Minnesota (a vertically integrated health care organization), functioning in a competitive managed care environment, has been implementing a hypertension services program. The program involves a team approach to care, with emphasis on patient participation in treatment; decentralized care delivery by nurse coordinators at primary care practice sites; ongoing training and education for patients and providers; and the continuous monitoring and evaluation of patient outcomes and satisfaction. JOB-LEVEL ISSUES: A variety of issues, such as the role and responsibilities of the nurse coordinator, became evident as the program moved towards operational status at four primary care practice sites, which prolonged the implementation period. PROCESS-LEVEL ISSUES: Issues relating to work process changes were more complicated to resolve and required, in some cases, changes in the proposed model. The most significant process-level issues related to educating physicians about the program to secure their participation and support. ORGANIZATION-LEVEL ISSUES: Such issues, which were the most difficult for program implementors to anticipate and resolve, included an organizational culture that emphasized decision making autonomy at primary practice sites. In part, the difficulty encountered in resolving organization-level issues reflected the implementors' lack of awareness of the strength or complexity of the environmental pressures facing the organization, as well as a lack of sensitivity to nuances relating to organizational culture. MOVING AHEAD: Two groups of hypertensive patients--at the implementation and comparison sites--will be compared with respect to satisfaction with care, clinical outcomes, and costs. Expansion of the model to patients with other chronic conditions is under consideration. PMID- 9407265 TI - Perceived threat and perceived control as predictors of the degree of fear in physical and social situations. AB - Seventy-one nonclinical subjects indicated their degree of anxiety or fear in response to five physical threat situations and four social threat situations. In addition, for each situation, they rated the probability and consequences of danger or threat generally associated with each situation and the degree of perceived control that they believed they would personally have in each situation. Multiple regressions indicated that only general probability of threat predicted fear in physical situations while the general consequences of threat and the degree of personal control predicted fear in response to social situations. PMID- 9407264 TI - Setting thresholds for MDS (Minimum Data Set) quality indicators for nursing home quality improvement reports. AB - BACKGROUND: Determining meaningful thresholds to reinforce excellent performance and flag potential problem areas is critical for quality improvement reports. Without thresholds, an organization may interpret its performance as superior to others because it is "better than average" and falsely assume it does not have care problems in certain areas. SETTING THRESHOLDS: The Minimum Data Set (MDS) assessment instrument is mandated for use nationwide in all nursing homes participating in Medicaid or Medicare programs. Since 1993 a research team at the University of Missouri-Columbia has been developing and testing quality indicators (QIs) derived from MDS data as a foundation for quality improvement activities. In July 1996, a cross-section of 13 clinical care personnel from nursing homes participated on an expert panel for threshold setting for QIs derived from MDS assessment data. Panel members individually determined good and poor threshold scores for each QI, reviewed statewide distributions of MDS QIs, and, two weeks later, completed a follow-up Delphi round. Three members of the research team reviewed the results of the expert panel and set the final thresholds. With thresholds established for good and poor scores, MDS QI scores are reported to a sample of Missouri nursing homes using the thresholds. CONCLUSIONS: To ensure that thresholds reflect current practice, threshold setting with another panel of experts will be repeated as needed, but at least biannually. The report format will be revised on the basis of user input, and a statewide study testing different educational support methods for quality improvement using MDS QIs is now underway. PMID- 9407266 TI - Mother-child agreement on self-report of anxiety in abused children. AB - This study investigates the lack of agreement in maternal and child report of child anxiety with a sample of abused and nonabused clinic-referred children. Based on the literature, it was predicted that nonabused clinic-referred children would report more symptoms of anxiety than their mothers would report for them. It was also predicted that mothers of abused children would report greater anxiety symptoms for their children than the children's self-report. Finally, it was predicted that maternal psychopathology, specifically anxiety, would increase the probability that mothers would overreport their children's anxiety. Mother child agreement based on anxiety symptoms assessed by the Quay Behavior Problem Checklist and the Revised-Children's Manifest Anxiety Scale was obtained on 54 male and female outpatients, 5 to 16 years of age. Overall, mothers reported significantly more anxiety for their children, than the children's own self report, irrespective of abuse history. The implications of the findings are discussed with respect to the validity of maternal and child report. Directions for future research are also offered. PMID- 9407267 TI - The neuropsychology of trichotillomania. AB - The pathophysiology of trichotillomania (TM) is not well understood. Overlap with obsessive compulsive disorder (OCD) has been proposed, although extant data are inconsistent in this regard. In the neuropsychological domain, some data have supported the proposed TM-OCD overlap. However, the available studies are limited in number, and they typically have sampled a restricted range of performance domains. To examine further neuropsychological functioning in TM, the present study compared performance of 21 patients with TM and 17 normal control (NC) participants on a broad battery of tests assessing intellectual functioning, auditory perception and language, visual perception, somatosensory function, motor ability, memory, concept formation, attention and information processing speed, impulsivity, and cerebral dominance. The TM group demonstrated poorer performance on all measures of divided, but not focused, attention. Correlational data suggested the potentially important role of negative affect in TM. Implications of the data for the conceptualization of TM are discussed. PMID- 9407268 TI - Computer-delivered modeling of exposure for spider phobia: relevant versus irrelevant exposure. AB - Spider phobic subjects (n = 45) completed 3 x 40 minute computer-delivered treatment sessions. Questionnaire ratings of phobic severity were completed pre- and posttreatment (n = 45) and 6-12 month follow-up (n = 38). The program used interactive animations to model self-exposure treatment methods. Subjects were randomized to one of three treatment groups (n = 15), each receiving a different version of the program. These treatment conditions were relevant exposure with feedback (REF), relevant exposure with no feedback (RENoF), or irrelevant exposure with feedback (IEF). Relevant exposure modeled exposure to spiders, irrelevant exposure to elevators. All groups showed significant phobic improvement following the treatment, as measured on a variety of instruments. Learning on the programs was demonstrated by a significantly increased performance (time taken to reach a target score) within each group across the three computer treatment sessions. Outcome was not significantly affected either relevance of exposure or the manipulation of the onscreen feedback. Subjects' report of exposure treatment undertaken at home correlated positively with phobic improvement. PMID- 9407269 TI - Prevalence of body dysmorphic disorder in patients with anxiety disorders. AB - Body Dysmorphic Disorder (BDD) is a debilitating disorder that often goes undetected in clinical practice. To provide information on the diagnostic correlates of BDD, we examined rates among outpatients seeking treatment for anxiety disorders. Participants (N = 165) were evaluated with a structured clinical interview and received the following primary diagnoses: panic disorder (n = 80), obsessive-compulsive disorder (n = 40), social phobia (n = 25) and generalized anxiety disorder (n = 20). Overall, 6.7% of patients met criteria for BDD. Rates were highest for social phobia (12%). When comorbid social phobia was excluded, rates of BDD were 1.5% in panic disorder, 6.7% in generalized anxiety disorder, and 7.7% in obsessive-compulsive disorder. In all cases, onset of social phobia preceded onset of BDD. Our findings draw attention to the prevalence of BDD in patients with social phobia. The potential etiologic significance of our findings is discussed. PMID- 9407270 TI - Personality disorders do not influence the results of cognitive and behavior therapy for obsessive compulsive disorder. AB - This study examined whether categorical or dimensional personality disorder variables affected treatment outcome in a sample of 52 patients with obsessive compulsive disorder who followed a standardized cognitive behavior therapy program. Treatment consisted of 12 weekly sessions and was completed by 43 patients. The Structured Clinical Interview for DSM-III-R personality disorders (SCID-II) was taken before the start of treatment by an independent rater. The treatment outcome measures included questionnaires and a Behavioral Assessment Test. Measurements were taken before and after treatment, and at 1 and 6 month follow-up tests. After the first follow-up test, further treatment was provided if clinically indicated. Neither categorical, nor dimensional personality disorder variables affected treatment outcome significantly. The inclusion of drop-outs in the analyses, did not change these results. Therefore, patients with obsessive compulsive disorder and concomitant personality disorder pathology should not be excluded from cognitive or behavior therapy for their obsessive compulsive complaints. Attributing therapy failure to concomitant Axis II pathology should be approached with caution. PMID- 9407271 TI - Sexual victimization, generalized perception of control, and posttraumatic stress disorder symptom severity. AB - We examined the relations among various characteristics of sexual victimization, posttraumatic stress disorder (PTSD) symptom severity, and generalized perception of control. Our main focus lay in testing three predictions derived from the animal model of PTSD articulated by Foa, Zinbarg, and Olasov-Rothbaum (1992) based on the effects of uncontrollable and/or unpredictable aversive events. A sample of 117 female undergraduates participated and completed self-report measures of past experience with child sexual abuse, adult sexual victimization, PTSD symptom severity, and locus of control. The results showed that child sexual abuse experienced on multiple occasions was associated with diminished generalized perception of control and that diminished generalized perception of control is associated with greater PTSD symptom severity following adult sexual victimization when experienced on a single occasion or involving force. These results provide partial support for the uncontrollability/unpredictability model of PTSD. Further research is necessary, however, to firmly establish the direction of causality involved in these associations. PMID- 9407273 TI - Heat-induced changes in the mechanics of a collagenous tissue: isothermal free shrinkage. AB - We present data from isothermal free-shrinkage tests (i.e., performed in the absence of mechanical loads) wherein bovine chordae tendineae were subjected to temperatures from 65 to 85 degrees C for 120 to 1200 s. These data reveal four new insights into heat-induced denaturation of a collagenous tissue. First, a characteristic time for the free shrinkage appears to exhibit an Arrhenius-type relationship with temperature. Second, scaling the actual heating time via the characteristic time results in a single correlation between free shrinkage and the duration of heating; this correlation suggests a time-temperature equivalence. Third, it is the cumulative, not current, heating time that governs the free shrinkage. And fourth, heat-induced free shrinkage is partially recovered when the tissue is returned to 37 degrees C, this recovery also being time-dependent. Although these findings will help guide future experimentation and constitutive modeling, as well as the design of new heat-based clinical therapies, there is a pressing need to collect additional isothermal data, particularly in the presence of well-defined mechanical loads. PMID- 9407272 TI - How does interoceptive exposure for panic disorder work? An uncontrolled case study. AB - To examine the influence of interoceptive exposure (IE) when used alone in the treatment of Panic Disorder (PD), 17 PD patients were presented with six IE sessions, using 35% CO2 as the exposure medium. The data indicate that IE alone is effective in reducing panic, panic-related fears, and general anxiety. However, the positive effects of IE do not appear to extend to agoraphobia, related fears, or depressed mood. Two distinct within-session patterns of fear response to IE were noted, one indicating habituation and the other indicating a lack of fear reduction. Although both patterns were associated with reductions in panic and anxiety following IE, the Habituators appeared to have a more positive outcome, which occurred more rapidly. These data suggest that IE may operate via two different pathways. Implications for understanding fear reduction are discussed, along with directions for future study. PMID- 9407274 TI - An in vitro osteotomy method to expose the medial compartment of the human knee. AB - This study was conducted to validate a new in vitro method to expose the medial compartment of the knee to be used in subsequent studies aimed at examining the load bearing capabilities of medial meniscal allografts. The new method involves an osteotomy and reattachment of the medial femoral condyle. The primary hypothesis was that the new method does not alter tibio-femoral contact pressure and area. To validate this method, the baseline contact pressure of the intact medial compartment was measured using a new nondestructive procedure for inserting pressure measurement film into the intact medial hemijoint. A secondary and related hypothesis was that incising the coronary ligament, a destructive method used by previous investigators to position pressure measurement film, alters the normal tibio-femoral contact pressure. To test these hypotheses, Fuji Prescale pressure-sensitive film was used to measure both tibio-femoral contact pressure and area within the medial compartment of the (1) intact knee, (2) the knee after osteotomizing and reattaching the medial femoral condyle, and (3) the osteotomized knee with an incised coronary ligament, using seven cadaver specimens. Measurements were taken at a compressive load of approximately two times body weight with the knee in 0, 15, 30, 45 deg of flexion. No significant differences between the intact and osteotomized knee were detected. Likewise, no significant differences were observed between the osteotomized knee and the osteotomized knee with an incised coronary ligament. These results confirm the utility of the new method in exposing the medial compartment for manipulation and placement of medial meniscal allografts in future studies examining the load bearing characteristics of meniscal allografts. PMID- 9407275 TI - Intraoperative measurement and biomechanical modeling of the flexor carpi ulnaris to-extensor carpi radialis longus tendon transfer. AB - Sarcomere length was measured intraoperatively in five patients undergoing tendon transfer of the flexor carpi ulnaris (FCU) to the extensor carpi radialis longus (ECRL) for radial nerve palsy. All measurements were made with the elbow in 20 deg of flexion. Prior to tendon transfer, FCU sarcomere length ranged from 2.84 +/- .12 microns (mean +/- SEM) with the wrist flexed to 4.16 +/- .15 microns with the wrist extended. After transfer into the ECRL tendon, sarcomere length ranged from 4.82 +/- .11 microns with the wrist flexed (the new longest position of the FCU) to 3.20 +/- .09 microns with the wrist extended, resulting in a shift in the sarcomere length operating range to significantly longer sarcomere lengths (p < 0.001). At these longer sarcomere lengths, the FCU muscle was predicted to develop high active tension only when the wrist was highly extended. A biomechanical model of this tendon transfer was generated using normative values obtained from previous studies of muscle architectural properties, tendon compliance, and joint moment arms. Predicted sarcomere lengths pre- and post tendon transfer agreed well with intraoperative experimental measurements. The theoretical wrist extension moment-wrist joint angle relationship was also calculated for a variety of values of FCU muscle length. These different lengths represented the different conditions under which the FCU could be sutured into the ECRL tendon. Variation in FCU muscle length over the range 200 mm to 260 mm resulted in large changes in absolute peak moment produced as well as the angular dependence of peak moment. This was due to the change in the region of FCU operation on its sarcomere length-tension curve relative to the magnitude of the ECRL moment arm. These data demonstrate the sensitivity of a short-fibered muscle such as the FCU to affect the functional outcome of surgery. In addition, we demonstrated that intraoperative sarcomere length measurements, combined with biomechanical modeling provide the surgeon with a powerful method for predicting the functional effect of tendon transfer surgery. PMID- 9407276 TI - A structurally based stress-stretch relationship for tendon and ligament. AB - We propose a mechanical model for tendon or ligament stress-stretch behavior that includes both microstructural and tissue level aspects of the structural hierarchy in its formulation. At the microstructural scale, a constitutive law for collagen fibers is derived based on a strain-energy formulation. The three dimensional orientation and deformation of the collagen fibrils that aggregate to form fibers are taken into consideration. Fibril orientation is represented by a probability distribution function that is axisymmetric with respect to the fiber. Fiber deformation is assumed to be incompressible and axisymmetric. The matrix is assumed to contribute to stress only through a constant hydrostatic pressure term. At the tissue level, an average stress versus stretch relation is computed by assuming a statistical distribution for fiber straightening during tissue loading. Fiber straightening stretch is assumed to be distributed according to a Weibull probability distribution function. The resulting comprehensive stress stretch law includes seven parameters, which represent structural and microstructural organization, fibril elasticity, as well as a failure criterion. The failure criterion is stretch based. It is applied at the fibril level for disorganized tissues but can be applied more simply at a fiber level for well organized tissues with effectively parallel fibrils. The influence of these seven parameters on tissue stress-stretch response is discussed and a simplified form of the model is shown to characterize the nonlinear experimentally determined response of healing medial collateral ligaments. In addition, microstructural fibril organizational data (Frank et al., 1991, 1992) are used to demonstrate how fibril organization affects material stiffness according to the formulation. A simplified form, assuming a linearly elastic fiber stress versus stretch relationship, is shown to be useful for quantifying experimentally determined nonlinear toe-in and failure behavior of tendons and ligaments. We believe this ligament and tendon stress-stretch law can be useful in the elucidation of the complex relationships between collagen structure, fibril elasticity, and mechanical response. PMID- 9407277 TI - A poroelastic model that predicts some phenomenological responses of ligaments and tendons. AB - Experimental evidence suggests that the tensile behavior of tendons and ligaments is in part a function of tissue hydration. The models currently available do not offer a means by which the hydration effects might be explicitly explored. To study these effects, a finite element model of a collagen sub-fascicle, a substructure of tendon and ligament, was formulated. The model was microstructurally based, and simulated oriented collagen fibrils with elastic orthotropic continuum elements. Poroelastic elements were used to model the interfibrillar matrix. The collagen fiber morphology reflected in the model interacted with the interfibrillar matrix to produce behaviors similar to those seen in tendon and ligament during tensile, cyclic, and relaxation experiments conducted by others. Various states of hydration and permeability were parametrically investigated, demonstrating their influence on the tensile response of the model. PMID- 9407278 TI - A large deformation biomechanical model for pressure ulcers. PMID- 9407279 TI - Optimal seat suspension design based on minimum "simulated subjective response". AB - This work addresses a method for improving vertical whole body vibration isolation through optimal seat suspension design. The primary thrusts of this investigation are: (1) the development of a simple model that captures the essential dynamics of a seated human exposed to vertical vibration, (2) the selection and evaluation of several standards for assessing human sensitivity to vertical vibration, and (3) the determination of the seat suspension parameters that minimize these standards to yield optimal vibration isolation. Results show that the optimal seat and cushion damping coefficients depend very much on the selection of the vibration sensitivity standard and on the lower bound of the stiffnesses used in the constrained optimization procedure. In all cases, however, the optimal seat damping obtained here is significantly larger (by than a factor of 10) than that obtained using existing seat suspension design methods or from previous optimal suspension studies. This research also indicates that the existing means of assessing vibration in suspension design (ISO 7096) requires modification. PMID- 9407280 TI - The use of inverse dynamics solutions in direct dynamics simulations. AB - Previous attempts to use inverse dynamics solutions in direct dynamics simulations have failed to replicate the input data of the inverse dynamics problem. Measurement and derivative estimation error, different inverse dynamics and direct dynamics models, and numerical integration error have all been suggested as possible causes of inverse dynamics simulation failure. However, using a biomechanical model of the type typically used in gait analysis applications for inverse dynamics calculations of joint moments, we produce a direct dynamics simulation that exactly matches the measured movement pattern used as input to the inverse dynamic problem. This example of successful inverse dynamics simulation demonstrates that although different inverse dynamics and direct dynamics models may lead to inverse dynamics simulation failure, measurement and derivative estimation error do not. In addition, inverse dynamics simulation failure due to numerical integration errors can be avoided. Further, we demonstrate that insufficient control signal dimensionality (i.e., freedom of the control signals to take on different "shapes"), a previously unrecognized cause of inverse dynamics simulation failure, will cause inverse dynamics simulation failure even with a perfect model and perfect data, regardless of sampling frequency. PMID- 9407281 TI - Shear properties of human brain tissue. AB - The objective of this study was to determine a relationship between shear stress and strain for human brain tissue by performing transient, single-pulse, high rate, shear displacement tests. A constant velocity, parallel plate shear test device was designed and fabricated. This equipment generated constant rate shear strains in cylindrical tissue samples mounted between the shear plates. The transverse reaction force at the upper end of the sample was measured during the event with a sensitive quartz piezoelectric force transducer, thus obtaining the force associated with the displacement versus time ramp. Shear tests were performed on 125 tissue samples taken from twelve fresh cadaver brain specimens. The average true shear stress and finite strain were calculated. A nonlinear, viscoelastic, standard solid model was fit to the constant rate test data and the material constants were determined. PMID- 9407282 TI - Axial impact biomechanics of the human foot-ankle complex. AB - Recent epidemiological, clinical, and biomechanical studies have implicated axial impact to the plantar surface of the foot to be a cause of lower extremity trauma in vehicular crashes. The present study was conducted to evaluate the biomechanics of the human foot-ankle complex under axial impact. Nine tests were conducted on human cadaver below knee-foot-ankle complexes. All specimens were oriented in a consistent anatomical position on a mini-sled and the impact load was delivered using a pendulum. Specimens underwent radiography and gross dissection following the test. The pathology included intra-articular fractures of the calcaneus and/or the distal tibia complex with extensions into the anatomic joints. Impactor load cell forces consistently exceeded the tibial loads for all tests. The mean dynamic forces at the plantar surface of the foot were 7.7 kN (SD = 4.3) and 15.1 kN (SD = 2.7) for the nonfracture and fracture tests, respectively. In contrast, the mean dynamic forces at the proximal tibial end of the preparation were 5.2 kN (SD = 3.1) in the nonfracture group, and 10.2 kN (SD = 1.5) in the fracture group. The foot and tibial end forces were statistically significantly different between these two groups (p < 0.01). The present investigation provides fundamental data to the understanding of the biomechanics of human foot-ankle trauma. Quantifying the effects of other factors such as gender and bone quality on the injury thresholds is necessary to understand foot ankle tolerance fully. PMID- 9407283 TI - Experimental investigation of the distribution of residual strains in the artery wall. AB - Arterial wall stresses are thought to be a major determinant of vascular remodeling both during normal growth and throughout the development of occlusive vascular disease. A completely physiologic mechanical model of the arterial wall should account not only for its residual strains but also for its structural nonhomogeneity. It is known that each layer of the artery wall possesses different mechanical properties, but the distribution of residual strain among the different mechanical components, and thus the true zero stress state, remain unknown. In this study, two different sets of experiments were carried out in order to determine the distribution of residual strains in artery walls, and thus the true zero stress state. In the first, collagen and elastin were selectively eliminated by chemical methods and smooth muscle cells were destroyed by freezing. Dissolving elastin provoked a decrease in the opening angle, while dissolving collagen and destroying smooth muscle cells had no effect. In the second, different wall layers of bovine carotid arteries were removed from the exterior or luminal surfaces by lathing or drilling frozen specimens, and then allowing the frozen material to thaw before measuring residual strain. Lathing material away from the outer surface caused the opening angle of the remaining inner layers to increase. Drilling material from the inside caused the opening angle of the remaining outer layers to decrease. Mechanical nonhomogeneity, including the distribution of residual strains, should thus be considered as an important factor in determining the distribution of stress in the artery wall and the configuration of the true zero stress state. PMID- 9407284 TI - Passive stress-strain measurements in the stage-16 and stage-18 embryonic chick heart. AB - The first stress-strain measurements on embryonic cardiovascular tissue are described here, obtained from cyclic uniaxial loading of the primitive ventricle. An excised ventricular segment from Hamburger/Hamilton stage-16 or stage-18 chicks (2-1/2 and 3 days of a 21-day incubation period) was mounted longitudinally between two small wires in oxygenated Krebs-Henseleit cardioplegia solution. One wire was attached to an ultrasensitive force transducer and the other to a Huxley micromanipulator controlled by remote motor drive. A real-time video tracking system calculated three myocardial surface strains based on the positions of three surface markers while the heart was deformed in a triangular wave pattern. Force transducer output was filtered, digitally sampled, and stored with strains and time. Results were plotted as strain (longitudinal, circumferential, shear, and principal) versus time, stress versus time, and stress versus longitudinal strain. The stress-strain curves were nonlinear, even at low strain levels. The hysteresis loops were large; mean hysteresis energy as a proportion of total cycle stored strain energy was 36 percent (stage 16) and 41 percent (stage 18). We created a finite element model of the ventricle and fit the model behavior to the experimental behavior to determine parameters for a stage-18 pseudoelastic strain-energy function of exponential form. The calculated exponential parameter is significantly lower than that found in corresponding uniaxial studies of mature myocardium, possibly indicating the lower fiber content of the immature tissue. The results of this study are the first step in characterizing material properties for comparisons with later developmental stages and with impaired and altered myocardium. The long-term goal is to aid in identifying the biomechanical factors regulating growth and morphogenesis. PMID- 9407285 TI - Computational approach for probing the flow through artificial heart devices. AB - Computational fluid dynamics (CFD) has become an indispensable part of aerospace research and design. The solution procedure for incompressible Navier-Stokes equations can be used for biofluid mechanics research. The computational approach provides detailed knowledge of the flowfield complementary to that obtained by experimental measurements. This paper illustrates the extension of CFD techniques to artificial heart flow simulation. Unsteady incompressible Navier-Stokes equations written in three-dimensional generalized curvilinear coordinates are solved iteratively at each physical time step until the incompressibility condition is satisfied. The solution method is based on the pseudocompressibility approach. It uses an implicit upwind-differencing scheme together with the Gauss Seidel line-relaxation method. The efficiency and robustness of the time-accurate formulation of the numerical algorithm are tested by computing the flow through model geometries. A channel flow with a moving indentation is computed and validated by experimental measurements and other numerical solutions. In order to handle the geometric complexity and the moving boundary problems, a zonal method and an overlapped grid embedding scheme are employed, respectively. Steady-state solutions for the flow through a tilting-disk heart valve are compared with experimental measurements. Good agreement is obtained. Aided by experimental data, the flow through an entire Penn State artificial heart model is computed. PMID- 9407286 TI - Enhancement in the effective thermal conductivity in rat spinotrapezius due to vasoregulation. AB - This study was undertaken to gain a better understanding of the countercurrent heat exchange of thermally significant blood vessels in skeletal muscle by measuring the vascular structure and flow in an exteriorized rat spinotrapezius muscle and estimating the enhancement in the effective thermal conductivity of the muscle. Detailed anatomic measurements of the number density and length of countercurrent vessel pairs between 45 and 165 microns diameter were obtained. Moreover, diameter and blood flow in the 1A to 3A vessels were measured for muscles in which pharmacological vasoactive agents were introduced, allowing one to vary the local blood flow Peclet number from 1 to 18 in the major feeding arteries. These combined measurements have been used to estimate the range of possible enhancement in the effective thermal conductivity of the tissue. The newly derived conduction shape factor in Zhu et al. for countercurrent vessels in two-dimensional tissue preparations was used in this analysis. Our experimental data indicated that the value of this conduction shape factor was about one-third to two-thirds the value for two countercurrent vessels of the same size and spacing in an infinite medium. The experiment also revealed that the Weinbaum Jiji expression for keff was valid for the spinotrapezius muscle when the largest vessels were less than 195 microns diameter. A fivefold increase in keff was predicted for 195 microns diameter vessels. Vasoregulation was also shown to have a dramatic effect on keff. A tissue that exhibits only small increases in keff due to countercurrent convection in its vasoconstricted state can exhibit a more than fivefold increase in keff in its vasodilated state. PMID- 9407287 TI - Oxygen mass transfer calculations in large arteries. AB - The purpose of this study was to model the transport of oxygen in large arteries, including the physiologically important effects of oxygen transport by hemoglobin, coupling of transport between oxygen in the blood and in wall tissue, and metabolic consumption of oxygen by the wall. Numerical calculations were carried out in an 89 percent area reduction axisymmetric stenosis model for several wall thicknesses. The effects of different boundary conditions, different schemes for linearizing the oxyhemoglobin saturation curve, and different Schmidt numbers were all examined by comparing results against a reference solution obtained from solving the full nonlinear governing equations with physiologic values of Schmidt number. Our results showed that for parameters typical of oxygen mass transfer in the large arteries, oxygen transport was primarily determined by wall-side effects, specifically oxygen consumption by wall tissue and wall-side mass transfer resistance. Hemodynamic factors played a secondary role, producing maximum local variations in intimal oxygen tension on the order of only 5-6 mmHg. For purposes of modeling blood-side oxygen transport only, accurate results were obtained through use of a computationally efficient linearized form of the convection-diffusion equation, so long as blood-side oxygen tensions remained in the physiologic range for large arteries. Neglect of oxygen binding by hemoglobin led to large errors, while arbitrary reduction of the Schmidt number led to more modest errors. We conclude that further studies of oxygen transport in large arteries must couple blood-side oxygen mass transport to transport in the wall, and accurately model local oxygen consumption within the wall. PMID- 9407288 TI - Measurements of airway dimensions and calculation of mass transfer characteristics of the human oral passage. AB - This paper presents measurements of the geometric shape, perimeter, and cross sectional area of the human oral passage (from oral entrance to midtrachea) and relates them through dimensionless parameters to the depositional mass transfer of ultrafine particles. Studies were performed in two identical replicate oral passage models, one of which was cut orthogonal to the airflow direction into 3 mm elements for measurement, the other used intact for experimental measurements of ultrafine aerosol deposition. Dimensional data were combined with deposition measurements in two sections of the oral passage (the horizontal oral cavity and the vertical laryngeal-tracheal airway) to calculate the dimensionless mass transfer Sherwood number (Sh). Mass transfer theory suggests that Sh should be expressible as a function of the Reynolds number (Re) and the Schmidt number (Sc). For inhalation and exhalation through the oral cavity (O-C), an empirical relationship was obtained for flow rates from 7.5-30.0 1 min-1: Sh = 15.3 Re0.812 Sc-0.986 An empirical relationship was likewise obtained for the laryngeal tracheal (L-T) region over the same range of flow rates: Sh = 25.9 Re0.861 Sc 1.37 These relationships were compared to heat transfer in the human upper airways through the well-known analogy between heat and mass transfer. The Reynolds number dependence for both the O-C and L-T relationships was in good agreement with that for heat transfer. The mass transfer coefficients were compared to extrathoracic uptake of gases and vapors and showed similar flow rate dependence. For gases and vapors that conform to the zero concentration boundary condition, the empirical relationships are applicable when diffusion coefficients are taken into consideration. PMID- 9407289 TI - Peristaltic transport of two-layered power-law fluids. AB - Peristaltic transport of two-layered power-law fluids in axisymmetric tubes is studied. Use of the power-law fluid model permits independent choice of shear thinning, shear thickening, or Newtonian fluids for the core and the peripheral layer. The interface between the two layers is determined from a transcendental equation in the core radius. The variation of the time-mean flow Q with the pressure rise or drop over one wavelength delta p is studied. It is observed that a negative time-mean flow is achieved under free pumping (delta p = 0) for the wave forms considered here if one of the peripheral layer and core fluids is non Newtonian. The rheology of the peripheral layer fluid is a dominant factor in producing a negative or positive mean flow. It is noticed that a sinusoidal wave always yields a positive mean flow for power-law fluids. The trapped bolus volume for sinusoidal peristaltic wave is observed to decrease with an increase in the rate of shear thinning of the core and the peripheral layer fluids. PMID- 9407290 TI - Monte Carlo model for determination of the role of heat generation in laser irradiated tissue. AB - A Monte Carlo model is described for modeling photo propagation in a scattering medium. The fraction of locally absorbed photons is proportional to the local rate of heat generation in laser-irradiated tissue and the associated distribution of light (fluence rate) is obtained by dividing the rate of heat generation by the local absorption coefficient. Examples of computed distributions of the rate of heat generation are presented for situations where light scattering in tissue is important. The method is applied to analyze treatment of Port Wine Stain and the selection of laser wavelengths for cyclophotocoagulation. PMID- 9407291 TI - Chronology of alveolar healing following immediate implantation of Ricinus communis polyurethane resin: histometric analysis in rats. AB - The purpose of the present study was to determine whether granules of Ricinus communis polyurethane resin implanted immediately after tooth extraction interfere with the time course of alveolar wound healing in rats. Progressive bone neoformation in parallel to a decrease in the volume fraction of connective tissue was quantified by a histometric method 1, 2, 3, and 6 weeks after tooth extraction. In spite of the biocompatible nature, the presence of polyurethane resin granules in the cervical third led to a small (9-22%) but significant delay in bone formation in the middle and apical alveolar thirds from the second week on, as compared to controls. PMID- 9407292 TI - On the magnetic behavior of the MA 956 superalloy. AB - MA 956 superalloy is a ferritic stainless material which develops a fine, dense, and well-adhered alpha-alumina layer upon heat treatment at elevated temperatures. This unique capability makes MA 956 attractive for surgical implants. In this work, the magnetic behavior of the material before and after thermal oxidation treatment required to develop the alumina layer is investigated. The thermal oxidation treatment yields a microstructure of elongated grains and a significant change in the texture. Despite these strong microstructural differences between the as-received and heat-treated materials, the hysteretic behavior is not greatly affected by them. MA 956 is a soft magnetic material irrespective of the material condition. The coercive force and residual magnetization of the material are somewhat lower under heat-treated conditions than in the as-received condition. PMID- 9407293 TI - Tissue response to fast-setting calcium phosphate cement in bone. AB - Fast-setting calcium phosphate cement (FSCPC) is a promising new bioactive cement with a significantly short setting time (approximately 5-6 min) compared to conventional calcium phosphate cement (c-CPC) (30-60 min) at physiologic temperatures. As a result of its ability to set quickly, it is applicable in surgical procedures where fast setting is required. In this study, FSCPC was implanted in rat tibiae to evaluate tissue response and biocompatibility. FSCPC was converted to hydroxyapatite (HAP) in bone faster than c-CPC in the first 6 h. By 24 h, significant amounts of both FSCPC and c-CPC had been converted to HAP. The conversion of FSCPC into HAP further proceeded gradually, reaching 100% within 8 weeks. Infrared spectroscopic analysis disclosed the deposition of B type carbonate apatite, which is a biological apatite contained in human dentin or bone, on the surface of the FSCPC. Histologically, FSCPC showed a tissue response similar to that of c-CPC. A slight inflammatory reaction was observed in the soft tissue apposed to both cements in the early period, and new bone was formed along the surface of the FSCPC at the adjacent bone. However, no resorption of either cement by osteoclasts or macrophages was observed within 8 weeks. We conclude that FSCPC is superior to c-CPC in clinical applications in oral and maxillofacial, orthopedic, plastic, and reconstructive surgery, since it shows a faster setting time and higher mechanical strength in the early period that are required in these surgical procedures, as well as osteoconductivity and excellent biocompatibility similar to that of c-CPC. PMID- 9407294 TI - Effect of crosslinking agents on acrylic bone cements based on poly(methylmethacrylate). AB - Three different crosslinking agents were added to the monomer content of the bone cement formulation based on poly(methylmethacrylate), PMMA, in various concentrations, and their effects on the curing parameters and mechanical properties were determined. The different crosslinking agents were dimethacrylates containing a range of chain lengths and degrees of flexibility. For the parent formulation of the bone cement, PMMA powder was used, and the properties and kinetics of curing were compared for the same system with and without crosslinking agents. It was found that at low concentrations of crosslinking agents, the mechanical properties were superior but steadily decreased with increasing concentrations. Poly(ethyleneglycoldimethacrylate), EGDMA(400), even when used at very low concentrations, produced a steady improvement in the mechanical properties and could be used in cement formulations with a view to reducing creep and improving mechanical properties. PMID- 9407296 TI - Macrophage subpopulation differentiation by stimulation with biomaterials. AB - Macrophages were elicited by the subcutaneous implantation of ultra high molecular weight polyethylene (UHMWPE) for periods of 2, 7, and 14 days in rats. Exudates of varying volumes were produced that was comprised of granulocytes, monocytes, immature and mature macrophages, and T-lymphocytes. No B-lymphocytes were observed at any time periods. Cell types were identified by their granularity and positivity to the following antibodies: leucocyte common antigen (LCA, pan leucocyte); CD11b/c (macrophage/monocyte); CD5 (T-lymphocyte); CD45RA (B-lymphocyte); HIS48 (granulocyte); ED2 (mature macrophage); and MCP-1 (monocyte chemoattractant protein 1). Monocytes isolated from control rat blood demonstrated a size slightly larger than that of granulocytes but with less granularity. Their size and granularity were followed over increasing time periods. The macrophages elicited by UHMWPE showed a similar pattern, with the exception of an apparently highly granular subpopulation with volumes similar to that of granulocytes but significantly more granular. The granular macrophage subset had a very high degree of ED2 and MCP-1 positivity, and their proportion, compared with other macrophages, was greatest at 2 days. The high MCP-1 expression was accounted for by MCP-1 molecules bound to the surface of a small proportion of macrophages that were activated. It is postulated that this subpopulation was responsible for the synthesis of the MCP-1 and could indicate a mechanism by which monocytes are attracted to the site of an implanted material. PMID- 9407297 TI - Molecular anatomy of freeze-fractured ultra-high-molecular-weight polyethylene as determined by low-voltage scanning electron microscopy. AB - Morphological similarities between virgin ultra-high-molecular-weight polyethylene (UHMWPE) powder and debris retrieved from failed UHMWPE total joint implants motivated this study's objective: to establish the internal microstructural features of consolidated UHMWPE. Cylindrical specimens were cored from a gamma-irradiation-sterilized tibial component (extruded from GUR 415 resin), and then these specimens were freeze-fractured at high strain rates. Low voltage scanning electron microscopy was used to examine these surfaces. Two types of areas were observed. The first were uniform, homogeneous, and continuous with microridge structures (45-70 nm wide) and hillocks (0.1-0.3 microns in diameter). The second was nonhomogeneous and discontinuous with febrils (10-200 nm long), microridges, fenestra as small as 20 nm, and large crater-like structures (6-12 microns in diameter). Many of the submicronsized structures observed were similar to the structures observed in virgin powder, as well as those observed by others from wear debris retrieval studies. These data support the hypotheses that wear debris originates, in part, from structures originally present in the powder resin, and that these structures retain their identity throughout consolidation, machining, and in vivo wear, and are released into periprosthetic tissues as wear debris. PMID- 9407295 TI - A flow cytometric immunoassay to quantify adsorption of complement activation products (iC3b, C3d, SC5b-9) on artificial surfaces. AB - Crosslinked agarose microspheres and various polystyrene microspheres were analyzed for complement components after incubation with serum at 37 degrees C for times up to 2 h. Quantification involved direct flow cytometric analysis of the beads after the bound complement proteins were indirectly fluorescently tagged by use of a monoclonal antibody against a complement protein: C5b-9, iC3b, C3d, C4d, Bb, C3a, and C1q. Calibration with fluorescein microbead standards demonstrated that the membrane attack complex (SC5b-9) was surface bound on all surfaces and that the surface concentration gradually increased to levels as high as 0.5 micrograms/cm2. Further, the surface bound represented a substantial percentage of the total generated. The iC3b level on polystyrene beads rapidly reached 0.09 micrograms/cm2 and the C3d levels were an order of magnitude less. On agarose beads the iC3b levels continually rose to 0.17 micrograms/cm2 and, as before, the C3d levels were substantially lower. The surface concentration of C4d and Bb on both surfaces were significant but less than 1.0 ng/cm2. There was minimal evidence of C3a and C1q adsorption for any surface. Use of amino polystyrene beads moderately reduced the level of bound iC3b, C3d, and SC5b-9, whereas carboxylated beads reduced the levels by almost a factor of two. The appreciable amounts of iC3b and SC5b-9 consistently noted on the artificial surfaces tested in this paper suggests that for these two activation products in vitro analysis of material induced complement activation should also include surface analysis. PMID- 9407298 TI - Altered mechanical properties in aortic elastic tissue using glutaraldehyde/solvent solutions of various dielectric constant. AB - The extent to which elastic tissue can be crosslinked in aldehydes and the mechanism of such action is unresolved in the literature. We have used glutaraldehyde/solvent solutions of decreasing dielectric constant (phosphate buffer, methanol, 95% ethanol, n-propanol, n-butanol) to alter the mechanical properties of aortic elastic tissue obtained from autoclaved and CNBr-purified bovine aortae. Treated and untreated hoop samples were examined for stress-strain and stress relaxation behavior and for residual stress using opening angle experiments as per Fung. The extent of exogenous crosslinking was analyzed through amino acid analysis. Mechanical properties of autoclaved elastic tissue varied with dielectric constant in glutaraldehyde/solvent treatments; however, solvent treatment alone produced no effect. Extensibility decreased with decreasing dielectric constant while tensile modulus changed over a range from 2.4% (-0.86 kPa) for glutaraldehyde/buffer to +35.3% (+14.3 kPa) for glutaraldehyde/n-propanol (untreated-treated). Residual stress experiments similarly showed a systematic decrease in opening angle with decreasing dielectric constant. Differences ranged from 10.5 degrees for glutaraldehyde/buffer to 22.2 degrees for glutaraldehyde/n-butanol. Interestingly, purification of aortae with CNBr reduced the effects of glutaraldehyde/n-butanol treatment. We hypothesize that CNBr differentially degraded the elastin-associated microfibrillar proteins in aortic elastic tissue, thus producing the observed differences in mechanical behavior. The observed phenomena in this study may be attributed to the composite structure of elastic tissue: elastin and microfibrillar protein. During treatment, conformational changes in elastin facilitated by polar/nonpolar interactions occurred which then were "locked" in by glutaraldehyde crosslinking of the microfibrillar proteins. By this mechanism the increases in both stiffness and time-dependent behavior observed after treatment may be explained. PMID- 9407299 TI - Proliferation and differentiation of human trabecular osteoblastic cells on hydroxyapatite. AB - In order to evaluate whether human osteoblastic cells differentiate normally on hydroxyapatite, we have compared the adhesion, proliferation, and differentiation of human trabecular (HT) osteoblastic cells on synthetic-dense hydroxyapatite and on standard plastic culture. We show here that initial HT cell attachment was 4 fold lower on hydroxyapatite than on plastic after 4 h of culture, and that normal cell attachment on hydroxyapatite was restored after 18 h of culture. HT cell proliferation was similar on the two substrates at 2-8 days of culture, but was lower on hydroxyapatite compared to plastic after 15 and 28 days of culture, as evaluated by DNA synthesis or cell number. HT cells cultured on both substrates produced an abundant extracellular matrix which immunostained for Type I collagen. The levels of carboxyterminal propeptide of Type I procollagen (P1CP) in the medium were lower in HT cell cultures on hydroxyapatite than on plastic. In addition, (3H)-proline incorporation into matrix proteins and the mean thickness of matrix layers were 52% and 26% lower, respectively, on hydroxyapatite compared to plastic after 4 weeks of culture, indicating that the total collagenous matrix synthesized by HT cells was lower on hydroxyapatite. However, (3H)-proline and calcium uptake expressed per cell was higher on hydroxyapatite than on plastic. The results show that human osteoblastic cells attach, proliferate, and differentiate on dense hydroxyapatite with a sequence similar to that of plastic. However, the growth of human osteoblastic cells is lower on hydroxyapatite in long-term culture, which results in a reduced amount of extracellular matrix, although matrix production per cell may be increased. PMID- 9407300 TI - Cytotoxic effects of acrylates and methacrylates: relationships of monomer structures and cytotoxicity. AB - Thirty-nine acrylates and methacrylates that had been used in dental resin materials were evaluated by a cytotoxicity test, and the relationships between their structures and cytotoxicity were studied to predict cytotoxic levels of dental resin materials in order to develop new low-toxic resin materials. All the acrylates evaluated were more toxic than corresponding methacrylates. In both the acrylates and methacrylates, a hydroxyl group seemed to enhance cytotoxicity. Dimethacrylates with 14 or fewer oxyethylene chains showed similar cytotoxicity while dimethacrylates with 23 oxyethylene chains showed lower cytotoxicity. The cytotoxicity ranking of monomers widely used in dental resin materials was bisphenol A bis 2-hydroxypropyl methacrylate (bisGMA) > urethane dimethacrylate (UDMA) > triethyleneglycol dimethacrylate (3G) > 2-hydroxyethyl methacrylate (HEMA) > methyl methacrylate (MMA). In acrylates, methacrylates, and ethylmethacrylates with either substituents, the lipophilicity of substituents affected their cytotoxicity, and an inverse correlation between IC50 and logP was observed. These results will be useful in developing new resin materials with low toxic monomer compositions. PMID- 9407301 TI - Synthetic scleral reinforcement materials. III. Changes in surface and bulk physical properties. AB - Changes in the physical properties of polymer materials during implantation in the biological environment can directly affect the ultimate performance of the polymer and/or device. We implanted four types of extraocular bands (porous, solid, composite, and patched) made from 11 types of materials in rabbit eyes and examined the changes in the physical strength and polymer structure of the implanted bands in terms of tensile strength measurements, creep analysis, and attenuated total internal reflectance--Fourier transform infrared spectroscopy (ATR-FTIR) at intervals up to 18 months after implantation. Most of the materials showed increases in tensile strength over the first 6 months in situ in the rabbit eye, followed by significant decreases between 6 and 18 months. Polymer bands that had been implanted for 18 months generally exhibited less creep behavior than unimplanted controls; for most of the bands, creep values ranged from 0% to 10% of the original length. ATR-FTIR of the solid bands and surface coatings showed protein deposition on all of the materials examined, with silicone materials and coatings least affected. Thirty-degree ATR-FTIR scans detected significant changes in the polymer structure for two of the band types: one solid (polyether urethane) and one porous (porous polyacrylate). In general, expanded polytetrafluoroethylene was the most stable in terms of tensile strength and creep. The least stable bands (composite bands made with porous polyurethane) were those that had undergone hydrolytic and/or oxidative degradation and chain scission of the polymer or alteration of the bond between the two materials making up the composite. These changes in physical properties and polymer structure observed after 18 months of implantation support the idea that polymer implant materials should be followed closely over several years in vivo to determine their suitability prior to use in humans. PMID- 9407302 TI - Adjuvancy effect of different types of silicone gel. AB - Women with silicone gel-filled breast implants (SBIs) are likely to be at a slightly higher risk of developing an autoimmune-like syndrome. This risk, although small, may be associated with the immunological adjuvancy property of the silicone gel. However, not all silicone gels are chemically formulated exactly the same and their adjuvancy behavior may vary. This study compared, in rats, the adjuvant effect of three different lots of silicone gel using ovalbumin (OVA) as the test antigen. Test bleeds were taken at 21, 48, 62, and 84 days post immunization and the rat sera were analyzed for anti-OVA antibodies by enzyme linked immunosorbent assay (ELISA). A delayed type hypersensitivity (DTH) test was performed on all the treated rats beginning at 14 post-immunization days. The results showed that silicone gel #3 (McGhan lot #S0400488) produced the highest mean anti-OVA antibody titer followed by silicone gel #1 (DC lot #HH019581) and silicone gel #2 (McGhan lot #DP9339). The DTH results showed that rats treated with silicone gel #1 and #3 had a clear positive response, whereas silicone gel #2 caused only a minimal response. These results demonstrate the immunological adjuvancy difference among three types of silicone gel. The chemical composition of each of these silicone gels, that would help explain these results, is yet to be determined. PMID- 9407303 TI - The effect of a subcutaneous silicone rubber implant with shallow surface microgrooves on the surrounding tissues in rabbits. AB - It has been suggested that during wound healing microtextured surfaces can alter events at the interface between implant surface surface and surrounding tissues. To investigate this phenomenon, smooth and microtextured silicone rubber implants were implanted subcutaneously in rabbits for 3, 7, 42, and 84 days. The textured implants possessed parallel surface microgrooves and ridges with a width of 2.0, 5.0, and 10.0 microns. All grooves had a depth of approximately 0.5 microns. SEM observation showed fibroblasts, erythrocytes, lymphocytes, macrophages, fibrin, and collagen on all implant surfaces after 3 and 7 days. After 42 and 84 days only little collagen, a small number of fibroblasts, but no inflammatory cells were seen on the implant surfaces. The fibroblasts were not oriented along the surface grooves on all textured surfaces. Three-dimensional reconstruction of CLSM images and LM images showed no significant differences between the thickness of the capsules surrounding the smooth and those surrounding the microgrooved implants. In contrast LM did show a significantly lower number of inflammatory cells and a significantly higher number of blood vessels in the capsules surrounding the microgrooved implants. Differences between the 2.0, 5.0, and 10.0 microns grooved implants were not detected. Our results concerning the capsule thickness suggest that the depth of our grooves was not sufficient to facilitate mechanical interlocking, but the cause for the observed differences in inflammatory response and number of blood vessels remains unclear. PMID- 9407304 TI - New anionic polyelectrolyte hydrogel for corneal surgery. AB - A new high-water-content (78%) anionic polyelectrolyte hydrogel was obtained by phase inversion (demixion) of a polymer solution containing 9.0% poly(acrylonitrile sodium methallylsulphonate), 85.0% dimethylformamide, and 6.0% saline solution (0.9% NaCl). The hydrogel is permeable to water, saline, urea, creatinine, glucose, human albumin, and saline-dissolved oxygen. Investigation of the interactions between human serum and surfaces prepared with the new yielded hydrogel, compared to serum interaction with silica-free silicone (RTV), regenerated cellulose (Cuprophan), MMA/PVP copolymer (Lidofilcon), PMMA (Perspex), FIFE (Gore-Tex), and poly(acrylonitrile sodium methallylsulphonate) hemodialysis membrane (AN-69), showed the hydrogel and hemodialysis membrane (both prepared with AN-69 copolymer) to be the only materials devoid of complement (C')-activating ability. PMID- 9407305 TI - Direct bone formation on alumina bead composite. AB - We have developed a composite (designated ABC), consisting of alumina bead powder as an inorganic filler and bisphenol-alpha-glycidyl methacrylate (Bis-GMA)-based resin as an organic matrix, which allows direct bone formation on its surface in vivo. Alumina bead powder was manufactured by fusing crushed alpha-alumina powder and quenching it. The beads took spherical form 3 microns in average size. According to powder X-ray diffraction and Fourier transform infrared spectroscopy, the alumina bead powder was composed of amorphous and delta-crystal phases of alumina in its main crystal structure. Fused-quenched silica glass filled composite (SGC) was used as a control. The proportion of filler added to the composites was 70% w/w. Mechanical testing of the ABC indicated that it would be strong enough for use under weight-bearing conditions. No apatite formation was detected on the surfaces of either composite after soaking in simulated body fluid for 28 days in vitro. Histological examination of rat tibiae for up to 8 weeks revealed that ABC bonded to bone directly via a layer of calcium, phosphorus, and alumina with no interposed soft-tissue layer. Moreover, the amount of bone directly apposed to the ABC surface increased with time, whereas with SGC there was poor direct bone formation even at 8 weeks. The precise mechanism of direct bone formation on ABC is as yet unknown but it is possible that changes in the crystallinity of alumina, which is known to be highly biocompatible, contribute to its excellent osteoconductivity in vivo. Although bioactive materials such as Bioglass or apatite and wollastonite-containing glass ceramic have previously been reported to form bone-like apatite on their surfaces under acellular conditions via simple chemical reactions, ABC does not have such characteristics, and presenting favorable conditions for osteoconduction and tissue calcification may lead to direct bone formation on its surface in vivo. PMID- 9407306 TI - Use of photografting technique for the immobilization of blood-grouping antibodies onto microtiter plates. AB - Commercial anti-A, anti-B, anti-A,B and anti-D monoclonal and polyclonal antisera were immobilized onto polystyrene microtiter plates using a photografting technique, to set up a new solid-phase assay (SPA) to be used for blood grouping. The reactivity and specificity of each grafted antisera were studied using red blood cells (RBCs) expressing normal and weak antigens. The stability of immobilized antisera was also studied. After dry storage of plates at +4 degrees C, room temperature, and +37 degrees C, SPA was performed using fresh and/or frozen RBCs. The same test was carried out after storing plates under protective conditions. Concordance of collected with expected results was obtained in all cases when the SPA was performed using monoclonal antisera and RBCs, with normal or weak expression of ABO and D antigens immediately after plate preparation or after dry storage at +4 degrees C. Plates stored dry at room temperature or at +37 degrees C gave inconsistent results, whereas a slight increase in reactivity was observed after storage under protective conditions. The specificity and the reactivity of tested antibodies were not modified by the immobilization procedure, not even after dry storage at +4 degrees C. Damage produced by water evaporation during dry storage in hard conditions could be reduced by adding a protective solution to microtiter wells at the beginning of storage. PMID- 9407307 TI - Significance of porosity and compliance of microporous, polyurethane-based microarterial vessel on neoarterial wall regeneration. AB - The microporous structure of artificial vascular grafts, which increases compliance and porosity simultaneously, may enhance neoarterial regeneration. In order to differentiate these effects, three models of segmented polyurethane grafts (inner diameter, 1.5 mm; wall thickness, 100 microns) with or without micropores fabricated using an excimer laser ablation technique, were prepared, and their neoarterial regenerative potentials were studied upon implantation: Model I (microporous, permeable, compliant); Model II (smooth-surfaced, impermeable, compliant); and Model III (smooth-surfaced, impermeable, noncompliant). In Models I and II, the pore or groove size (diameter, 100 microns) and pore or groove arrangement were fixed, and consequently their compliances were almost identical. Irrespective of model, the luminal surfaces were coated with benzophenone-derivatized gelatin and subsequently photocured. Twenty grafts (length, 20 mm) of each model were implanted in the aortas of rats. Predetermined implantation periods were 4, 12, and 24 weeks. Total patency rate decreased in the order Model I (100%), II (87%), and III (59%) grafts. All patent grafts were completely endothelialized after 12 weeks of implantation, irrespective of model. After 12- and 24-week implantations, in Model I grafts, the neoarterial wall was thin, and smooth muscle cells (SMCs) were of the contractile phenotype. In Model II grafts, the neoarterial wall exhibited considerable thickening. In Model III grafts, the neoarterial wall exhibited marked thickening, and SMCs were of the synthetic phenotype. The neoarterial wall thickness at the midportion of the grafts after 24 weeks of implantation increased in the order Model I (48 +/- 8 microns), II (146 +/- 87 microns), and III (385 +/- 21 microns) grafts. These results strongly suggest that compliance matching and porosity synergistically resulted in neoarterial wall restoration without appreciable thickening. PMID- 9407308 TI - Molecular modeling study of adsorption of poly-L-lysine onto silica glass. AB - A research program was initiated with both experimental and computational chemistry based molecular modeling components to investigate specific amino acid surface interactions. The experimental portion of this study, with details reported elsewhere, investigated the adsorption of selected molecular weights of poly(L-lysine) onto silica glass microspheres with the adsorption enthalpy per adsorbed mer determined to be -0.23 +/- 0.13 kcal/mol (mean +/- 95% confidence interval). Molecular modeling of this system was then conducted using two approaches: an AM1 semiempirical molecular orbital method to predict L lysine/glass interaction energy and an MM2 molecular mechanics method to investigate the structural configuration for poly(L-lysine). The modeling predicted a minimum energy configuration of a rotational backbone structure for poly(L-lysine) with approximately one full rotation occurring about every 8 mers, and that the amine side chains of the L-lysine will hydrogen bond with the silica surface with an average adsorption energy of approximately -0.34 kcal/mol/mer. The molecular modeling results are in good agreement with the experimentally measured value and provide insights into possible molecular-level behavior which would be very difficult to determine by experimental analyses alone. This work demonstrates the use of molecular modeling in conjunction with experimental studies to investigate complex molecular interactions. PMID- 9407309 TI - Magnetic field exposure enhances mRNA expression of sigma 32 in E. coli. AB - The mechanism of interaction between weak electromagnetic fields and cells is not understood. As a result, the health effect(s) induced by exposure to these fields remains unclear. In addition to questions relating to the site of initial magnetic field (MF) interactions, the nature of the cell's response to these perturbations is also unclear. We examined the hypothesis that the cells respond to MFs in a manner similar to other environmental stressors such as heat. Using the bacterium Escherichia coli, we examined the mRNA levels of sigma 32, a protein that interacts with RNA polymerase to help it recognize a variety of stress promoters in the cell. Our data show that the intracellular level of sigma 32 mRNA is enhanced following a 15-min exposure to a 60 Hz, 1.1 mT magnetic field. PMID- 9407310 TI - Regulation of connexin43 expression and function by prostaglandin E2 (PGE2) and parathyroid hormone (PTH) in osteoblastic cells. AB - Connexin43 (Cx43) forms gap junctions that mediate intercellular communication between osteoblasts. We have examined the effects of prostaglandin E2 (PGE2) and parathyroid hormone (PTH) on gap junctional communication in the rat osteogenic sarcoma cells UMR 106-01. Incubation with either PGE2 or PTH rapidly (within 30 min) increased transfer of negatively charged dyes between UMR 106-01 cells. This stimulatory effect lasted for at least 4 h. Both PGE2 and PTH increased steady state levels of Cx43 mRNA, but only after 2-4 h of incubation. Transfection with a Cx43 gene construct linked to luciferase showed that this effect of PTH was the result of transcriptional upregulation of Cx43 promoter. Stimulation of dye coupling and Cx43 gene transcription were reproduced by forskolin and 8Br-cAMP. Exposure to PGE2 for 30 min increased Cx43 abundance at appositional membranes in UMR 106-01, whereas total Cx43 protein levels increased only after 4-6 h of incubation with either PGE2 or PTH. Inhibition of protein synthesis by cycloheximide did not affect this early stimulation of dye coupling, but it significantly inhibited the sustained effect of PTH and forskolin on cell coupling. In summary, both PTH and PGE2, presumably through cAMP production, enhance gap junctional communication in osteoblastic cell cultures via two mechanisms: initial rapid redistribution of Cx43 to the cell membrane, and later stimulation of Cx43 gene expression. Modulation of intercellular communication represents a novel mechanism by which osteotropic factors regulate the activity of bone forming cells. PMID- 9407311 TI - Extracellular matrix modulates mesangial cell apoptosis and mRNA expression of cathepsin-B and tissue transglutaminase. AB - Mesangial matrix is a dynamic structure which modulates mesangial cell function. Since accumulation of matrix precedes the development of focal glomerulosclerosis, we studied the effect of different matrices on mesangial cell (MC) apoptosis. Suspended mesangial cells became apoptotic in a time dependent manner. Collagen type III did not modulate MC apoptosis when compared to cells grown on plastic. MCs grown on Matrigel, collagen type I and IV showed an increased number of apoptotic cells when compared to MCs grown on plastic. DNA end-labeling further confirmed these observations. MCs grown on Matrigel showed enhanced (P < 0.05) mRNA expression for tissue transglutaminase (TTG) and cathepsin-B. Mesangial cells grown on Matrigel also showed enhanced expression of superoxide dismutase (SOD). We conclude that mesangial cells require attachment to the matrix for their survival and alteration of the quality of matrix modulates mesangial cell apoptosis. PMID- 9407312 TI - Apoptosis during bone-like tissue development in vitro. AB - We present evidence of cell death by apoptosis during the development of bone like tissue formation in vitro. Fetal rat calvaria-derived osteoblasts differentiate in vitro, progressing through three stages of maturation: a proliferation period, a matrix maturation period when growth is downregulated and expression of the bone cell phenotype is induced, and a third mineralization stage marked by the expression of bone-specific genes. Here we show for the first time that cells differentiating to the mature bone cell phenotype undergo programmed cell death and express genes regulating apoptosis. Culture conditions that modify expression of the osteoblast phenotype simultaneously modify the incidence of apoptosis. Cell death by apoptosis is directly demonstrated by visualization of degraded DNA into oligonucleosomal fragments after gel electrophoresis. Bcl-XL, an inhibitor of apoptosis, and Bax, which can accelerate apoptosis, are expressed at maximal levels 24 h after initial isolation of the cells and again after day 25 in heavily mineralized bone tissue nodules. Bcl-2 is expressed in a reciprocal manner to its related gene product Bcl-XL with the highest levels observed during the early post-proliferative stages of osteoblast maturation. Expression of p53, c-fos, and the interferon regulatory factors IRF-1 and IRF-2, but not cdc2 or cdk, were also induced in mineralized bone nodules. The upregulation of Msx-2 in association with apoptosis is consistent with its in vivo expression during embryogenesis in areas that will undergo programmed cell death. We propose that cell death by apoptosis is a fundamental component of osteoblast differentiation that contributes to maintaining tissue organization. PMID- 9407313 TI - Interaction of the transcription factor Sp1 with the nuclear pore protein p62 requires the C-terminal domain of p62. AB - The transcription factor Sp1 plays an important role in the expression of many cellular genes. In studies of proteins that associate with Sp1, a 62-kDa glycoprotein was found in immunoprecipitates of Sp1. This protein was detected in these immunoprecipitates by the monoclonal antibody, RL2, which was originally raised against nuclear pore proteins but was subsequently found to recognize an epitope that contains O-linked N-acetylglucosamine (O-GlcNAc). The association of this protein with Sp1 could be blocked by SDS denaturation of the protein complex. Western blot analysis of the Sp1 immunoprecipitate using antibodies to p62 nucleoporin indicated that this nuclear pore protein associates with Sp1. Furthermore, immunoprecipitation of p62 nucleoporin resulted in the coprecipitation of Sp1. Recombinant p62, expressed as a GST-fusion protein using a vaccinia virus system, also interacted with both recombinant and native Sp1. This interaction between p62 and Sp1 required the C-terminus of p62 and the C terminus was able to bind Sp1, albeit less efficiently than native p62. A mammalian two-hybrid interaction assay was devised in which p62 was fused to the Gal4 DNA-binding domain. This system also indicated that p62, through its C terminus, interacts with Sp1 in the living cell. We propose that this interaction of a nuclear pore protein with Sp1 may reflect the nuclear organization required to bring transcribable DNA in contact with the transcription factors. PMID- 9407314 TI - Somatostatin increases mitogen-induced IL-2 secretion and proliferation of human Jurkat T cells via sst3 receptor isotype. AB - The neuropeptide somatostatin (SRIF) modulates normal and leukemia T cell proliferation. However, neither molecular isotypes of receptors nor mechanisms involved in these somatostatin actions have been elucidated as yet. Here we show by using RT-PCR approach that mitogen-activated leukemia T cells (Jurkat) express mRNA for a single somatostatin receptor, sst3. This mRNA is apparently translated into protein since specific somatostatin binding sites (K11 = 78 +/- 3 pM) were detected in semipurified plasma membrane preparations by using 125I-Tyr1-SRIF14 as a radioligand. Moreover, somatostatin inhibits adenylyl cyclase activity with similar efficiency (IC50 = 23 +/- 4 pM) thus strongly suggesting a functional coupling of sst3 receptor to this transduction pathway. The involvement of sst3 receptor in immuno-modulatory actions of somatostatin was assessed by analysis of neuropeptide effects on IL-2 secretion and on proliferation of mitogen-activated Jurkat cells. Our data show that in the concentrations comprised between 10 pM and 10 nM, somatostatin potentiates IL-2 secretion. This effect is correlated with somatostatin-dependent increase of Jurkat cell proliferation since the EC50 concentrations for both actions were almost identical (EC50 = 22 +/- 9 pM and EC50 = 12 +/- 1 pM for IL-2 secretion and proliferation, respectively). Altogether, these data strongly suggest that in mitogen-activated Jurkat cells, somatostatin increases cell proliferation through the increase of IL-2 secretion via a functional sst3 receptor negatively coupled to the adenylyl cyclase pathway. PMID- 9407315 TI - Desquamin is an epidermal ribonuclease. AB - Desquamin is a glycoprotein that we have isolated from the upper granular layer and the stratum corneum of human epidermis; it is not ordinarily expressed in submerged cultures, whose terminal differentiation stops short of formation of these layers. The exogenous addition of desquamin to human cultured keratinocytes extended their maturation, and hematoxylin staining indicated a loss of cell nuclei. For confirmation, cultured cells were lysed in situ, and the nuclei were incubated with desquamin for several days, then stained with hematoxylin. Damage to the nuclei was evident: the nuclear inclusions remained intact, while the surrounding basophilic nuclear matrix was degraded. Desquamin was then tested directly for nuclease activity. Ribonuclease activity was determined by incubating desquamin with human epidermal total RNA and monitoring the dose dependent disappearance of the 28S and 18S ribosomal RNA bands in an agarose/formaldehyde gel. On RNA-containing zymogels, we confirmed the RNase activity to be specific to desquamin. Using synthetic RNA homopolymers, we found the active RNase domains to be limited to cytosine residues. On the contrary, DNA was not degraded by an analogous procedure, even after strand-separation by denaturation. PMID- 9407316 TI - Extracellular pH modulates the activity of cultured human osteoblasts. AB - The effect of medium pH on the activity of cultured human osteoblasts was investigated in this study. Osteoblasts derived from explants of human trabecular bone were grown to confluence and subcultured. The first-pass cells were incubated in Hepes-buffered media at initial pHs adjusted from 7.0 to 7.8. Osteoblast function was evaluated by measuring lactate production, alkaline phosphatase activity, proline hydroxylation, DNA content, and thymidine incorporation. Changes in medium pH were determined from media pHs recorded at the beginning and end of the final 48 h incubation period. As medium pH increased through pH 7.6, collagen synthesis, alkaline phosphatase activity, and thymidine incorporation increased. DNA content increased from pH 7.0 to 7.2, plateaued from pH 7.2 to 7.6, and increased again from pH 7.6 to 7.8. The changes in the medium pH were greatest at pHs 7.0 and 7.8, modest at pHs 7.4 and 7.6, and did not change at 7.2, suggesting that the pHs are migrating towards pH 7.2. Lactate production increased at pH 7.0 but remained constant from 7.2 to 7.8. These results suggest that in the pH range from 7.0-7.6 the activity of human osteoblasts increases with increasing pH, that this increase in activity does not require an increase in glycolytic activity, and that pH 7.2 may be the optimal pH for these cells. PMID- 9407317 TI - Retinoic acid treatment elevates matrix metalloproteinase-2 protein and mRNA levels in avian growth plate chondrocyte cultures. AB - Matrix metalloproteinases (MMPs) play a crucial role in tissue remodeling. In growth plate (GP) cartilage, extensive remodeling occurs at the calcification front. To study the potential involvement of MMPs in retinoic acid (RA) regulation of skeletal development, we studied the effect of all-trans-RA on MMPs levels in mineralizing chicken epiphyseal chondrocyte primary cultures. When treated for 4 day periods on days 10 and 17, RA increased levels of an approximately 70 kDa gelatinase activity. The N-terminal sequence of the first 20 amino acid residues of the purified enzyme was identical to that deduced from chicken MMP-2 cDNA. Time-course studies indicated that RA elevated MMP-2 activity levels in the cultures within 16 h. This increase was inhibited by cycloheximide and was enhanced by forskolin. The increase in MMP-2 activity induced by RA was accompanied by an increase in MMP-2 mRNA levels and was abolished by treatment with cycloheximide. This upregulation of MMP levels by RA in GP chondrocytes is consistent with its effects on osteoblasts and osteosarcoma cells and opposite its inhibitory effects on fibroblasts and endothelial cells. It may well be related to the breakdown of the extracellular matrix in the GP and would be governed by the availability of RA at the calcification front where extensive vascularization also occurs. PMID- 9407318 TI - Selected nuclear LINE elements with mitochondrial-DNA-like inserts are more plentiful and mobile in tumor than in normal tissue of mouse and rat. AB - The nuclear DNA of normal and tumor mouse and rat tissue was examined for mitochondrial-DNA-like inserts by means of the Southern blot technique. The two probes were 32P-labeled cloned mitochondrial DNA. KpnI, which doesn't cut either mitochondrial DNA, was one of the restriction enzymes, while the enzymes that fragment mitochondrial DNA were for mouse and rat PstI and BamHI, respectively. When KpnI alone was used in the procedure a nuclear LINE family whose elements had mitochondrial-DNA-like insertions was selected. Such elements were much more abundant in tumor than in normal tissue. The results with PstI alone and BamHI alone and each combined with KpnI indicated that there were mobile LINE elements with mitochondrial-DNA-like inserts in the nuclear genome of tumor. The mouse tissues were normal liver and a transplantable lymphoid leukemic ascites cell line L1210 that had been carried for 40 years. The rat tissues were normal liver and a hepatoma freshly induced by diethylnitrosoamine in order to minimize the role of 40 years of transplantation. Our unitary hypothesis for carcinogenesis of 1971, which suggested these experiments, has been augmented to include mobile nuclear elements with inserts of mitochondrial-DNA-like sequences. Such elements have been related to diseases of genetic predisposition such as breast cancer and Huntington's disease. PMID- 9407319 TI - Induction and characterization of metallothionein in chicken epiphyseal growth plate cartilage chondrocytes. AB - Following exposure to cadmium or zinc, chickens were sacrificed and the liver, kidney, and bone epiphyseal growth plates harvested. When cytosolic extracts of the growth plate cartilage were fractionated by gel filtration chromatography, a protein with high metal-binding capacity and low ultraviolet (UV) absorbance eluted in the same position as liver metallothionein (MT) and a MT standard. Cd or Zn treatment resulted in a 25-fold or 5-fold induction in growth plate MT, respectively. In liver the greatest level of MT induction was seen with short term Cd exposures. In contrast, MT levels in the growth plate increased as the duration of Cd exposure increased. Induction of MT in growth plate chondrocyte cell cultures was observed for media Cd concentrations of > or = 0.1 microM and Zn concentrations of > or = 100 microM. Basal and inducible levels of MT declined through the culture period and were lowest in the terminally differentiated mineralized late stages of the culture. Alkaline phosphatase activity was also lowest in the late-stage cultures, while total cellular protein increased throughout the culture period. Treatment of chondrocytes with Zn prior to Cd exposure resulted in a protective induction of MT. Pre-treatment of chondrocytes with dexamethasone resulted in suppressed synthesis of MT upon Cd exposure and greater Cd toxicity. Both Cd and Zn resulted in significantly increased levels of MT mRNA in chondrocyte cell cultures. Dexamethasone treatment resulted in an approximate 2- to 3-fold increase in MT mRNA. This is contrary to the finding that MT protein levels were decreased by dexamethasone. The findings suggest that an increased rate of MT degradation in dexamethasone-treated and late-stage chondrocyte cultures may be associated with the terminally differentiated phenotype. PMID- 9407320 TI - Gene transfer of human heme oxygenase into coronary endothelial cells potentially promotes angiogenesis. AB - Heme oxygenase (HO-1) is a stress protein that has been suggested to participate in defense mechanisms against agents that induce oxidative injury such as hemoglobin/heme, hypoxia-ischemia and cytokines. Overexpression of HO-1 in endothelial cells (EC) might, therefore, protect against oxidative stress produced under these pathological conditions, by generation of CO, a vasodilator, and bilirubin, which has antioxidant properties that enhance blood vessel formation to counteract hypoxia-induced injury. A plasmid containing the cytomegalovirus promoter (pCMV) neomycin human HO-1 gene complexed to cationic liposomes, lipofectin, was used to transfect rabbit coronary microvessel EC. Cells transfected with human HO-1 gene demonstrated a twofold increase in HO activity and maintained a similar phenotype as in the nontransfected cells. Cell number in transfected cells with human HO-1 gene increased by about 45%, as compared to nontransfected or those transfected with control pCMV. Transfected and nontransfected EC revealed a similar response to basic fibroblast growth factor (bFGF) in capillary formation. However, transfected cells with the human HO-1 gene exhibited a twofold increase in blood vessel formation. The angiogenic response of EC to overexpression of HO-1 gene provides direct evidence that the inductive form of HO-1 following injury represents an important tissue adaptive mechanism for moderating the severity of cell damage produced in inflammatory reaction sites of hemorrhage, thrombosis and hypoxic-ischemia. Thus, HO-1 may participate in the regulation of EC activation, proliferation and angiogenesis. PMID- 9407321 TI - Changes in the activity of cdk2 and cdk5 accompany differentiation of rat primary oligodendrocytes. AB - Oligodendrocytes, the myelinating cells of the central nervous system, are terminally differentiated cells that originate through asynchronous waves of proliferation and differentiation of precursors present at birth. Withdrawal from cell cycle and onset of differentiation are tightly linked and depend on an intrinsic program modulated by the action of growth factors. p27 plays a central and obligatory role in the initiation of oligodendrocyte differentiation and cessation of proliferation. In this paper, we have characterized the role of modulation of cdk2 and cdk5 kinase activity during the process of oligodendrocyte precursor differentiation. As rat primary oligodendrocytes differentiate in culture there is a fall in cdk2 activity and a rise in cdk5 activity as well as an increase in the cdk inhibitor, p27 protein. The decline in cdk2 activity is not accompanied by a drop in cdk2 protein level, suggesting that it results from inhibition of cdk2 activation rather than decreased protein expression. Taken together, these data suggest that oligodendrocytes may withdraw from the cell cycle at G1-S transition through inactivation of cdk2 activity, possibly initiated by increasing amount of p27, and that cdk5 may have a role until now unrecognized in the differentiation of oligodendrocytes. PMID- 9407322 TI - The value of hepatitis C virus genotyping to epidemiological and clinical studies. PMID- 9407323 TI - Distribution of hepatitis C virus genotypes among blood donors in Taiwan. AB - The aim of this study was to investigate the prevalence and distribution of hepatitis C virus (HCV) genotypes of blood donors in Taiwan. RNA was extracted from the serum of anti-hepatitis C virus-positive carriers and this was followed by reverse transcriptase-polymerase chain reaction (RT-PCR) using type-specific primers for the presence of HCV genotypes, 1a, 1b, 2a, 2b, 3a and 6a. Of the 604 anti-HCV-positive specimens, the PCR demonstrated that 93.0% (562/604) were positive for at least one HCV genotype. The remaining 42 specimens (7%) were HCV negative. Among the 562 HCV-positive specimens, 505 (89.8%) contained HCV 1a, 1b, 2a, 2b and 3a as the only genotype, with a prevalence of 0.4% (2/562), 60.1% (338/562), 15.5% (87/562), 11.9% (67/562), and 2.0% (11/562), respectively. No HCV genotype 6a was found. Thirty-seven specimens (6.6%) exhibited mixed infections with multiple HCV genotypes that included types 1b, 2a and 2b, while 20 (3.5%) HCV RNA-positive sera remained unclassified. These results confirm that the predominant HCV genotype in Taiwan is 1b. In addition, genotypes 1a and 3a can also be found in Taiwan at low frequency. PMID- 9407324 TI - Negative strand of hepatitis C virus RNA in the liver of patients with chronic hepatitis C after interferon treatment. AB - In patients receiving interferon therapy for chronic hepatitis C, serum hepatitis C virus (HCV) RNA often reverts from an undetectable to a detectable form after completion of treatment. Detection of the negative strand of HCV-RNA in liver tissue is regarded as an index of viral proliferation. Therefore, we investigated changes in the hepatic negative-strand HCV-RNA following interferon therapy to determine whether this parameter could predict the long-term response to treatment. The subjects of this study were 27 patients with chronic active hepatitis C. Serum positive-strand and hepatic tissue negative-strand HCV-RNA were detected using polymerase chain reaction. At the completion of interferon treatment, serum HCV-RNA was not detected in 21 patients. One year following treatment it remained undetectable in 14 of these patients but it had reverted to a detectable form in seven. The 14 patients in whom hepatic negative-strand RNA was not detected between 2 weeks and 12 months after treatment, had not relapsed after another year. In the 13 remaining patients, negative-strand RNA was found in liver tissue and serum RNA either reverted to a detectable form or remained detectable throughout. From these findings, we conclude that the detection of negative-strand HCV-RNA in liver tissue 2 weeks after the completion of interferon therapy is useful for predicting the long-term effect of therapy. PMID- 9407325 TI - Short-term effect of portal vein arterialization on hepatic protein synthesis and endotoxaemia after extended hepatectomy in dogs. AB - To assess the effect of partial portal arterialization on the remaining liver after usually lethal extended hepatectomy, 30 mongrel dogs underwent 84% partial hepatectomy and were divided into three groups as follows: group 1, 84% partial hepatectomy (n = 10); group 2, 84% partial hepatectomy and splenectomy (n = 10); group 3, 84% partial hepatectomy and splenectomy and splenic artery-vein (A-V) shunt (n = 10). Another five dogs were pre-operatively killed normal controls. Portal vein flow (PVF) decreased to about 60% in groups 1 and 2, but PVF in group 3 was maintained at the pre-operative level. Oxygen saturation of portal vein blood increased markedly, to between 83% (group 1) and 88% (group 3). Portal vein pressure (PVP) increased in groups 1 and 2 by 1.6 to 1.7 times the pre-operative value, but no significant difference in PVP, serum aspartate aminotransferase (AST) and arterial ketone body ratio was found between the three groups. Plasma endotoxin levels after 84% partial hepatectomy were significantly lower in group 3 than in groups 1 and 2. Both of hepatocellular and secretory protein synthesis were enhanced in group 3 compared with the other two groups. These results suggest that partial portal arterialization using a splenic A-V shunt might bring about a beneficial effect on remaining liver function after extended hepatectomy. PMID- 9407326 TI - Case report: congenital absence of the portal vein associated with nodular hyperplasia in the liver. AB - Congenital absence of the terminal portion of the portal vein with visceral venous return to the suprahepatic inferior vena cava, a rare malformation, was demonstrated in an 18-year-old Japanese woman. She had nodular hyperplasia in the liver and a non-functioning pancreatic endocrine tumour. It is generally believed that reduction of portal venous flow causes atrophic changes and, subsequently, nodular hyperplasia occurs in a well-perfused area in the liver. However, the liver was not perfused by the portal vein in this case. It is suggested that nodular hyperplasia can occur without portal blood flow. PMID- 9407327 TI - Abnormality in fatty acid composition of gastric mucosal phospholipids in patients with liver cirrhosis and its correction with a polyunsaturated fatty acid-enriched soft oil capsule. AB - The abnormal composition of the fatty acids in gastric mucosal phospholipids was examined in 11 patients with non-alcoholic liver cirrhosis accompanied by gastritis and in 10 healthy subjects without liver disease and gastritis. The cases positive for serum anti-Helicobacter pylori immunoglobulin G antibody were excluded. The arachidonic acid (AA, C20:4n-6) contents of the two main components of phospholipid, the phosphatidylcholine (PC) and phosphatidylethanolamine (PE) fractions, in gastric biopsy specimens were significantly lower in the cirrhosis group, although PC and PE contents in gastric biopsy specimens were not altered. In the cirrhosis group, AA contents of the PC fractions in gastric biopsy specimens were significantly correlated with AA contents of plasma PC fractions and serum albumin levels. Soft oil capsules containing polyunsaturated fatty acid (PUFA) such as AA, eicosapentanoic acid (EPA, C20:5n-3) and docosahexanoic acid (DHA, C20:6n-3) were administered after each meal daily for 4 weeks to six cirrhotic patients and four control subjects, who were randomly selected from the patients and control subjects. After administration of PUFA-enriched oil capsules, gastric mucosal AA contents of PC and PE fractions were significantly improved almost to the control levels. In three cirrhotic patients with severe or mild gastritis whose gastric mucosal lesions were endoscopically shown to have responded to PUFA-enriched oil capsules, AA contents of plasma PC and PE fractions before administration were much lower than in the remaining three cirrhotics that did not respond to the PUFA-enriched oil capsules, but significantly improved to the control levels after administration of oil capsules. The results demonstrate that administration of PUFA-enriched soft oil capsules increased AA contents of the phospholipids in gastric mucosa and thus improved gastric mucosal lesions endoscopically in cirrhotic patients with gastritis. PMID- 9407328 TI - Management of Helicobacter pylori. European and North American guidelines. PMID- 9407329 TI - Racial differences in Helicobacter pylori seroprevalence in Singapore: correlation with differences in peptic ulcer frequency. AB - The aim of this study was to determine, first, whether racial differences exist in the seroprevalence of Helicobacter pylori infection in Singapore, and second, whether these differences correlate with racial differences in peptic ulcer frequency. A commercial serological test for immunoglobulin (Ig)G antibody to H. pylori which was 90% sensitive and 83% specific in our population was used to screen 403 adult blood donors of Chinese, Malay and Indian origin, aged between 15-60 years. Serum specimens from 84 paediatric patients admitted to the Paediatrics Department, National University of Singapore, with non gastroenterological illnesses were also tested. In all three races, seroprevalence of H. pylori increased with age. Indians have the highest prevalence of infection followed by Chinese and Malays. Peptic ulcer prevalences are known to be highest in Chinese, followed by Indians and Malays. The Malays have the lowest prevalence of H. pylori and peptic ulcer among the three races in Singapore. Indians have a higher prevalence of H. pylori antibodies but a lower frequency of peptic ulcer than the Chinese. Racial differences in peptic ulcer frequency between Chinese and Indians are not explained by the prevalence of H. pylori infection; other environmental or genetic factors may be involved. PMID- 9407330 TI - Relationship between local immune response to Helicobacter pylori and the diversity of disease: investigation of H. pylori-specific IgA in gastric juice. AB - In order to evaluate the relationship between local immune response to Helicobacter pylori and the diversity of disease, 77 asymptomatic subjects who underwent a health examination were studied. Helicobacter pylori-specific IgG in serum and H. pylori-specific IgA in gastric juice were measured by ELISA, and the measured IgA titre was classified into two grades, low or high. Histological classification of gastritis was performed according to the Sydney system. Cytokines in gastric juice were also measured, and the cytotoxin-associated gene A (cagA) status of H. pylori was tested by PCR. Of the 65 subjects who were positive for H. pylori-specific IgG in serum, 38 (58.5%) were classified as H. pylori-specific IgA low titre in gastric juice and 27 (41.5%) had high titres. In the IgG-positive, IgA-low group, the rate of peptic ulcers (especially duodenal ulcers) in endoscopic findings was higher (P < 0.05); the score of activity and the density of H. pylori were higher (P < 0.001 and P < 0.05, respectively); the score of metaplasia was lower (P < 0.05); and the level of interleukin-1 beta was lower (P < 0.05) than in the IgG-positive, IgA-high group. The positive rate of the cagA gene was 84.4% and there was no significant difference between the two groups. There were differences in endoscopic and histological findings between the IgG-positive, IgA-low and the IgG-positive, IgA-high groups. It is suggested that persons infected with H. pylori can be divided into two different states of disease according to local immune response. PMID- 9407331 TI - Virulence-associated genes as markers of strain diversity in Helicobacter pylori infection. AB - To establish a marker of strain diversity of Helicobacter pylori, a genetic examination was performed based on the detection rates by PCR of cagA and vacA, which are known to be virulence-associated genes. The test strains were obtained from 70 patients suffering from gastric ulcer (GU), 82 patients with duodenal ulcer (DU) and 48 patients with gastritis (GS). Fragments located in the three different regions of vacA were amplified; V1 being the upstream portion, V2 the mid-portion and V3 the downstream portion. For cagA, the detection rates were 70% for GU, 79% for DU and 50% for GS, showing a significantly higher rate for DU than for GS (P = 0.0005). With V1, the detection rates were 90% for GU, 90% for DU and 69% for GS, giving a significantly higher rate for GU than for GS (P = 0.0036) and also giving a significantly higher rate for DU than for GS (P = 0.0019). With V2, the detection rates were 60% for GU, 70% for DU and 44% for GS, giving a significantly higher rate for DU than for GS (P = 0.0024). The differences in vacA gene polymorphism were closely related to the evidence of gastroduodenal ulcers in H. pylori infection. Furthermore, the detection rates of cagA and polymorphisms of vacA by PCR could be used as markers of strain diversity in H. pylori-induced gastroduodenal ulcer. PMID- 9407332 TI - Review: the surgical management of ulcerative colitis. AB - The treatment of ulcerative colitis requires careful review of the medical and surgical options. The surgical procedure of choice is proctocolectomy with ileal pouch-anal anastomosis. This procedure removes the diseased mucosa, effectively curing the disease whilst maintaining the normal route of defecation and continence. Other surgical options that may be considered in selected patients include proctocolectomy with either a Brooke ileostomy or a Kock pouch, and abdominal colectomy with ileorectal anastomosis. The choice of operation requires consideration of the advantages and disadvantages of a particular procedure and must be tailored to an individual patient's needs and circumstances. PMID- 9407333 TI - Thiolmethyltransferase activity in the human colonic mucosa: implications for ulcerative colitis. AB - Ulcerative colitis is associated with a selective reduction of n-butyrate oxidation by the colonic epithelial cells although the reason for this has been unclear. Colonic epithelial cell n-butyrate oxidation can be inhibited in vitro by incubation with sulphide but the role of mucosal detoxification of sulphide in the metabolic welfare of the colonic mucosa has not been examined. This study aimed to assess the role mucosal detoxification of sulphide by thiolmethyltransferase (TMT)-mediated methylation may play in protecting the healthy colonic mucosa from the adverse effects of luminal sulphide. Colonic epithelial cell suspensions from healthy human proximal (n = 9) and distal colon (n = 10) were incubated in the presence of 14C-labelled n-butyrate (5 mmol/L) alone, butyrate plus sodium hydrogen sulphide (NaHS) (1.5 mmol/L), or butyrate plus NaHS plus S-adenosyl-methionine 1,4 butane disulphonate (SAMe) (5 mmol/L). Study end points were metabolic performance (14CO2 production) and mucosal TMT activity. Incubation with NaHS induced a significant inhibition of 14CO2 production compared with control incubations (P < 0.001) which was similar for proximal and distal colonic cell suspensions. S-adenosyl-methionine 1,4 butane disulphonate reversed this effect completely in proximal but not in distal cell incubations, suggesting a greater susceptibility of the distal colon to the sulphide effect. Although median whole mucosal TMT values did not differ between proximal and distal colonic mucosa, a non-normal distribution of distal TMT values was observed. However, neither the degree of sulphide inhibition of control 14CO2 production nor the degree to which SAMe reversed this inhibition correlated with whole mucosal TMT activity. The study concluded that regional variation exists in TMT activity in the human colon but whilst methylation appears to protect colonic epithelial cells against sulphide-induced inhibition of n-butyrate oxidation, this cannot be directly correlated with mucosal TMT activity. PMID- 9407334 TI - Serum levels of sCD23, interleukin-10 and interferon-gamma in patients with coeliac disease. AB - The role of cellular and humoral immunity coeliac disease was investigated by the measurement of serum levels of interleukin-10 (IL-10), interferon-gamma (IFN gamma) and soluble CD23 (sCD23). Coeliac disease was diagnosed by duodenal biopsy and response to a gluten-free diet (GFD). The results were compared with age and sex-matched patients with non-specific upper gastrointestinal symptoms and normal duodenal histology. While the levels of serum IL-10 were significantly elevated (P < 0.01) in patients with coeliac disease taken as a whole, the levels of serum IFN-gamma were normal and sCD23 significantly decreased (P < 0.002). The median serum sCD23 was significantly lower in the coeliac disease patients not on a GFD compared with those asymptomatic on a GFD (P < 0.03) and the control group (P < 0.0004). The coeliac disease patients on a GFD also had significantly lower serum sCD23 and higher IL-10 compared with the control group (P < 0.01 and P < 0.015). There was no significant difference in the serum IL-10 between the coeliac disease patients on a GFD and those not on a GFD and between the latter and the control group. The low levels of serum sCD23 in coeliac disease suggest diminished humoral immunity and, conversely, exaggerated cellular immunity. The aetiology of the raised levels of IL-10 in coeliac disease is unclear and similar to that observed in patients with inflammatory bowel disease. However, this may represent a regulatory response to the elevated levels of proinflammatory cytokines described in coeliac disease. A combination of diminished sCD23 and raised IL-10 is clearly unusual as both are associated with Th2-type functions. The possible causes of this finding are discussed. PMID- 9407335 TI - Oesophageal cancer associated with other primary cancers: a study of 31 patients. AB - The aim of this study was to investigate the characteristics of oesophageal cancer associated with other primary cancers and the survival rate after surgery for the patients with these cancers. Of 202 patients with oesophageal cancer treated in the Second Department of Surgery, Shinshu University School of Medicine between 1981 and 1995, 31 patients (15.3%) had oesophageal cancer associated with other primary cancers. Twenty-one synchronous and 10 metachronous associated cancers were found and 25 of them were resected. Early-stage oesophageal cancer was much more frequent in the associated cases than in the non associated cases. The stomach was the most frequently associated organ. The numbers of cases with triple and quadruple cancers were three and one, respectively. Three of these cases had intervals of over 6 years between tumours. Three cases with other primary cancers which had intervals of over 7 years after oesophagectomy were found, and two were carcinomas of the reconstructed gastric tube. In the outcome after surgery for oesophageal cancer, there was no difference between the associated and the non-associated cases, and also no difference between the synchronous and metachronous associated cases. Regarding the five-year and 10-year survival rates after surgery for the first cancers, the synchronous cases had a poorer outcome than did the metachronous cases. In conclusion, oesophageal cancer with other primary cancers is not always rare, and its outcome is not poor compared with that of the non-associated cases. These patients may achieve survival by early detection of both lesions and positive treatment. It is important to consider the risk of other primary cancers after oesophagectomy, and the success of the reconstructed gastric tube should be followed by endoscopy. PMID- 9407336 TI - Molecular biology of hepatitis B virus e antigen. AB - Hepatitis B virus (HBV) e antigen (HBeAg) was discovered in 1972 as one of the serological markers of HBV infection. Although 25 years have passed since its initial discovery, the function of this antigen in the life cycle of HBV has remained elusive. Mutations in the HBV genome that prevent the expression of HBeAg do not abolish the replication of HBV, indicating that this antigen is not essential for HBV replication. In contrast, the conservation of the HBeAg gene in the genomes of related animal viruses, including the distantly related duck HBV, argues for an important function of this antigen. The purpose of the present article is to review the molecular biology of HBeAg and to examine its possible functions in the life cycle of HBV. PMID- 9407337 TI - Molecular biology of hepatitis D virus: research and potential for application. AB - Superinfection by hepatitis D virus (HDV) leads to acute hepatitis and causes progression to liver cirrhosis in a significant proportion of hepatitis B surface antigen (HBsAg) carriers. Current regimens (interferon) to treat hepatitis D patients has only transient but no lasting effects. New approaches are, therefore, warranted. Recently, several laboratory studies have discovered interesting properties of HDV that may become targets for antiviral chemicals. Viral replication requires the small hepatitis delta antigen (s-HDAg). The s-HDAg is a nuclear phosphoprotein. There is evidence indicating that phosphorylation is important for HDV replication. A second step of replication requires HDV-RNA self cleavage and self-ligation. Interestingly, one group of antibiotics, the aminoglycosides, exerts strong suppression effects on HDV ribozyme activities. In the following stage of viral assembly, two post-translational modifications, namely isoprenylation of large HDAg and glycosylation of HBsAg are involved. Agents capable of blocking the two modifications should reduce viral production. These four possible targets are reviewed. For prevention, effective vaccines are not yet available. Two novel approaches are discussed. The first demonstrates the immunogenicity of a nucleic acid vaccine in mice. The second approach assembled an empty HDV particle in yeast. Advances on such laboratory investigations may provide new methods for the control of hepatitis D in the future. PMID- 9407339 TI - Histopathology and pathobiology of hepatotropic virus-induced liver injury. AB - The present report concerns current knowledge regarding immunopathogenesis that can be applied in the interpretation of histopathological changes in acute and chronic viral hepatitis. The histopathological features of viral hepatitis have not been changed and light microscopic examination remains essential for making a diagnosis and classification of chronic hepatitis and for the provision of objective parameters on grading and staging. However, new understanding and knowledge of viral pathogenesis, host immune responses, the biological behaviour of the causative viral agents and, in particular, viral interference in multiple hepatotropic viral infections must be taken into consideration in the interpretation of histopathological and immunopathological findings of liver tissues. This report also presents some histopathological analyses on multiple hepatotropic viral infections. It can be concluded that the diagnostic histological criteria for acute hepatitis remain applicable in such settings. However, the cause of acute flare up in chronic hepatitis could not be determined without clinical, virological and serological information. Routine histopathology cannot distinguish a new infection from an acute exacerbation due to a high level of viral replication or mutant virus. A repertoire of immunocytochemical stainings for viral antigens is helpful, but caution must be exercised in suggesting a specific viral aetiology due to the fact that suppression of pre existing viral antigens can be pronounced when the new or concurrent infection is hepatitis C virus related. PMID- 9407338 TI - Novel hepatitis agents: the significance of clinical and experimental studies. An overview. AB - A new virus with genomic organization similar to that of the family Flaviviridae was identified in patients with viral hepatitis and designated hepatitis G virus (HGV) or hepatitis GB virus C (GBV-C). HGV/GBV-C can be transmitted by blood and results in persistent infection, as shown in patients with posttransfusion non-A, non-B hepatitis, in transfusion recipients, and in donors of blood received by HGV-positive patients. The parenteral route of transmission of HGV/GBV-C infection was confirmed in experimentally infected chimpanzees. Epidemiologic studies of sporadic, community-acquired viral hepatitis have not indicated an association between HGV/GBV-C and acute non A-E hepatitis. Thus, the disease association of this new virus remains unconfirmed and its role in the etiology of acute and chronic hepatitis is unclear. The experimental model of HGV/GBV-C infection may define the biology of the virus replication. PMID- 9407340 TI - Natural history of chronic hepatitis B virus infection: an immunopathological study. AB - Three clinicopathological phases of chronic hepatitis B virus (HBV) infection are identified. First, is immune tolerance of HBV. High levels of viraemia are associated with normal alanine aminotransferase (ALT) levels and minimal histological lesions. More than 30-40% of hepatocytes have the hepatitis B core antigen (HBcAg), predominantly in their nuclei. Maternally derived hepatitis B e antigen (HBeAg) crossing the placenta may result in the elimination of T helper cells responsive to HBeAg/HBcAg. This phase can last for periods ranging from a few weeks to 10 or more years until the immune tolerance is lost. Second, is the immune clearance of HBV. Intermediate levels of viraemia are associated with fluctuating ALT levels and active and ongoing hepatitis. Approximately 20-30% of hepatocytes have HBcAg, predominantly in their cytoplasm. Expression of pre-core defective HBV mutants during chronic HBV infection may lead to a reduction in the secretion of HBeAg and may trigger the beginning of the immuno-elimination phase. The mechanism of intrahepatic shift of HBcAg from the nucleus to the cytoplasm and the decreased levels of viraemia in this phase may be, at least in part, secondary to liver damage and regeneration. Third, is latent infection with residual integrated HBV. Undetectable viraemia is associated with normal ALT levels and no virus-induced liver damage. With regard to hepatocyte expression of HBsAg in chronic HBV infection, membrane staining of HBsAg on hepatocytes has been shown to correlate well with the presence of viraemia. The degree of cytoplasmic hepatitis B surface antigen (HBsAg) expression inversely correlates with the level of viraemia. Therefore, HBsAg carriers with high levels of viraemia have low levels of cytoplasmic hepatitis B surface antigen (HBsAg) expression, while those with low levels of viraemia have high levels of cytoplasmic HBsAg expression. However, several exceptions have been identified. High levels of viraemia associated with high levels of cytoplasmic HBsAg expression were recognized in patients with fibrosing cholestatic hepatitis. In contrast, low levels of viraemia associated with low levels of cytoplasmic HBsAg expression were recognized in patients with hepatitis C virus but not hepatitis D virus superinfection. PMID- 9407341 TI - Fas system and apoptosis in viral hepatitis. AB - Hepatitis C virus (HCV) and hepatitis B virus (HBV) are the major causative agents of chronic liver disease. However, the mechanisms responsible for liver cell injury remain to be clarified. Playing crucial roles in the clearance of viral infection are cytotoxic T lymphocytes. Recently, it has been demonstrated that perforin- and Fas-based mechanisms account for all T cell-mediated cytotoxicity. Therefore, we examined the correlation between liver cell damage and the Fas system in the liver of patients with chronic hepatitis C. Fas is a cell surface protein that mediates apoptosis with treatment of the Fas ligand or the anti-Fas antibody. To investigate the role of Fas in type C hepatitis, we examined the correlation between liver cell damage and Fas expression. Fas expression was found mainly in the cytoplasm of hepatocytes and these positive cells were found particularly among infiltrating lymphocytes. A high prevalence of Fas expression was shown in liver tissue with more severe inflammation. The Fas system-mediated death signal requires the interaction of Fas ligand with Fas on target cells. We isolated a 1.9 kb cDNA clone for the human Fas ligand and examined the expression of the Fas ligand in liver-infiltrating mononuclear cells obtained from patients with chronic hepatitis C. The open reading frame encodes 281 amino acids. Next, we examined the expression of the Fas ligand in liver infiltrating mononuclear cells obtained from patients with chronic hepatitis C. The amplified products (231 bp) derived from Fas ligand transcripts were detected in liver-infiltrating mononuclear cells, whereas no signal was observed in liver tissues. In HCV infection, Fas expression in hepatocytes is up-regulated in accordance with the severity of liver inflammation. When HCV-specific T cells migrate into hepatocytes and recognize the viral antigen via the T cell receptor, they become activated and express Fas ligand that can transduce the apoptotic death signal to Fas-bearing hepatocytes. Thus, the Fas system plays an important role in liver cell injury by HCV infection. PMID- 9407342 TI - T cell mechanisms in the immunopathogenesis of viral hepatitis B and C. AB - Considerable evidence suggests that immune mechanisms are involved in the pathogenesis of both hepatitis B and C. Both CD4+ and CD8+ T cell responses to viral antigens are important mechanisms that may be responsible for the hepatocyte damage in hepatitis B and C. CD4+ T cell proliferative responses to hepatitis B core antigen (HBcAg) in terms of stimulation index are correlated with hepatitis activity. These responses can be demonstrated in both adult and paediatric patients, and are more vigorous in patients with acute self-limited hepatitis B than in patients with chronic hepatitis B. Patients with hepatitis C also had a significant CD4+ T cell response to hepatitis C virus (HCV) antigens. These responses are also vigorous in acute hepatitis C with recovery than in those cases that evolve to chronic hepatitis C. In terms of human leucocyte antigen (HLA) class I-restricted, CD8+ cytotoxic T lymphocyte (CTL) response, antigenic peptides derived from HBcAg, hepatitis B surface antigen (HBsAg), and polymerase have been demonstrated as the targets for CTL recognition in hepatitis B patients. Multiple CTL epitopes within both HBsAg and HBcAg can be detected by sensitizing target cells with synthetic peptides. Similar to hepatitis B virus (HBV) infection, multispecific, HCV-specific CTL responses can coexist with an extensive quasispecies of viral variants. The mechanisms of viral persistence in both hepatitis B and C are not yet clarified. PMID- 9407343 TI - Hepatitis in patients with end-stage renal disease. AB - Hepatitis B and hepatitis C are two common pathogens causing chronic hepatitis in patients with end-stage renal disease (ESRD). With the acceptance of hepatitis B s antigen (HBsAg) screening, infected patients have been identified and isolated over the past 20 years. Consequently, hepatitis B is now being seen less frequently in dialysis units. Even though hepatitis B has become less of a problem, non-A, non-B hepatitis has been recognized as a significant problem since 1979. With the availability of serological testing for hepatitis C virus (HCV), more specific information is now available in regard to HCV infection in dialysis patients. The prevalence of anti-HCV in haemodialysis (HD) patients is quite variable, ranging from 5 to over 50%. Anti-HCV positivity is associated with previous blood transfusions, mode of therapy and duration of haemodialysis. In Spain and Italy, the annual seroconversion rates of HCV antibodies in dialysis patients are 2-9%; this rate was much higher in Taiwan (15%). Whether patients with HCV infection should be identified and isolated during HD treatment is an issue of controversy. Transplantation is associated with increases in hepatitis B virus (HBV) replicative markers. The survival disadvantage in HBsAg-positive recipients usually did not become apparent until 8 years after transplantation. Hepatitis C virus-infected renal transplant recipients are presumably in a similar situation to patients with hepatitis B, although confirmatory data are currently lacking. Coinfection of HBV and HCV may lead to aggressive liver disease and cirrhosis. A hepatitis B vaccine is recommended for all susceptible dialysis patients. Dialysis patients have lower response rates to hepatitis B vaccines than do other people. Currently, no vaccine is available for hepatitis C. To date, there are no effective treatments available for hepatitis B and hepatitis C. Combination therapy with interferon/lamivudine for hepatitis B and interferon/ribavirin for hepatitis C may offer a promise of effective control of viral replication in the future. PMID- 9407344 TI - Drug-induced hepatic injury. AB - Drugs and other chemical toxins account for less than 5% of cases of jaundice or acute hepatitis and fewer cases of chronic liver disease, but they are an important cause of more severe types of hepatic injury. Drug reactions produce an array of hepatic lesions that mimic all known hepatobiliary diseases; this poses a diagnostic challenge for physicians and pathologists. Diagnosis of drug-induced hepatic injury is circumstantial, with positive rechallenge being the only factor that unequivocally implicates a particular agent. Nonetheless, other aspects of the temporal relationship between drug ingestion and adverse reaction, exclusion of other diseases, the presence of extrahepatic features of drug hypersensitivity and some findings on liver biopsy can lend support to the diagnosis. Some of these issues will be explored in this review by considering contemporary paradigms of drug-induced hepatic injury. Factors that predispose to dose dependent hepatic injury will be considered in relation to acetaminophen, an example of acute hepatotoxicity, and methotrexate, an agent that can produce hepatic fibrosis. Flucloxacillin will be discussed as an example of drug-induced cholestatic hepatitis often associated with prolonged cholestasis and the vanishing bile duct syndrome. Minocycline and diclofenac will be mentioned as two drugs for which drug hepatitis is an exceedingly rare complication. Finally, the evidence that Chinese herbal medicines can be hepatotoxic will be reviewed. PMID- 9407345 TI - Review: molecular approaches to inherited liver disease. Focus on Wilson disease. AB - Although infectious agents account for a large proportion of liver disease, inherited forms of liver disease, often treatable, must not be overlooked. New approaches to the cloning of disease genes are allowing an increased understanding of the basic defect of human diseases. Identification of disease genes can have practical applications for diagnosis, can provide information on prognosis and can lead to new therapies. This review focuses on selected inherited liver diseases and the knowledge to be gained from molecular studies. Three genetic diseases affecting the liver, Wilson disease, haemochromatosis and alpha 1-antitrypsin deficiency, are selected as examples of current approaches to the study of genetic liver disease. PMID- 9407346 TI - Recent developments in autoimmune liver diseases. AB - Several diseases are regarded as autoimmune liver diseases. Apart from the cholestatic liver diseases, primary biliary cirrhosis, primary sclerosing cholangitis, these include autoimmune hepatitis, hepatitis as part of the autoimmune polyendocrine syndrome type 1 (APS-1) and particular overlap syndromes such as autoimmune cholangitis (also called antimitochondrial antibody negative primary biliary cirrhosis [PBC]), overlap syndrome chronic active hepatitis (CAH)/PBC and the overlap syndrome primary sclerosing hepatitis (PSC)/CAH. In addition, auto-antibodies may be observed during the course of chronic viral hepatitis, in particular chronic hepatitis C and D. Finally, a small number of drug-induced liver diseases is immune mediated. The following article will review our recent progress in the field of autoimmune hepatitis including APS-1 and autoimmunity in viral hepatitis and immune-mediated drug-induced liver disease. PMID- 9407347 TI - Pathogenesis of alcoholic liver disease with particular emphasis on oxidative stress. AB - Oxidative stress is well recognized to be a key step in the pathogenesis of ethanol-associated liver injury. Ethanol administration induces an increase in lipid peroxidation either by enhancing the production of oxygen reactive species and/or by decreasing the level of endogenous antioxidants. Numerous experimental studies have emphasized the role of the ethanol-inducible cytochrome P450 in the microsomes and the molybdo-flavoenzyme xanthine oxidase in the cytosol. This review shows the putative role of ethanol-induced disturbances in iron metabolism in relation to iron as a pro-oxidant factor. Ethanol administration also affects the mitochondrial free radical generation. Many previous studies suggest a role for active oxygens in ethanol-induced mitochondrial dysfunction in hepatocytes. Recent studies in our laboratory in the Department of Internal Medicine, Keio University, using a confocal laser scanning microscopic system strongly suggest that active oxidants generated during ethanol metabolism produce mitochondrial membrane permeability transition in isolated and cultured hepatocytes. In addition, acetaldehyde, ethanol consumption-associated endotoxaemia and subsequent release of inflammatory mediators may cause hepatocyte injury via both oxyradical-dependent and -independent mechanisms. These cytotoxic processes may lead to lethal hepatocyte injury. Investigations further implicate the endogenous glutathione-glutathione peroxidase system and catalase as important antioxidants and cytoprotective machinery in the hepatocytes exposed to ethanol. PMID- 9407348 TI - Recent advances in the pathophysiology of portal hypertension. AB - Portal hypertension is a common complication of chronic liver disease. Increased resistance to portal blood flow through a cirrhotic liver initiates the development of portal hypertension. In addition, alteration of neural and humoral regulations, endothelins, and stellate cells all contribute to the increased intrahepatic resistance. Moreover, the collateral circulation represents an additional site of increased resistance to portal blood flow. Increased splanchnic blood flow appears to play an important role in the maintenance of chronic portal hypertension. Several mechanisms have been proposed to explain this haemodynamic derangement including increased circulating vasodilators, endothelial-derived vasodilators, and decreased vascular reactivity to vasoconstrictors. Finally, the development of portal hypertension induces peripheral arterial vasodilation. The arterial vasodilatation may result in an increase in vascular underfilling which in turn leads to sodium retention and plasma volume expansion. The increased plasma volume is necessary for the development of increased cardiac output and the full expression of hyperdynamic circulation in portal hypertension. PMID- 9407349 TI - Evoked potentials in liver diseases. AB - Evoked potentials are objective and quantitative methods capable of evaluating functions of both peripheral and central nervous systems (PNS and CNS). During the past 8 years, we have been using somatosensory, brainstem auditory, and pattern-reversal visual evoked potentials (SEP, BAEP, VEP) to study hepatic encephalopathy (HE) as well as functional status of the PNS and CNS in various liver diseases including viral hepatitis B, alcoholic liver disease and Wilson's disease (WD). In HE irrespective of its etiologies, there is a sequential prolongation and eventual disappearance of cortical components of the median nerve evoked SEP while there is no change in BAEP, suggesting that HE is primarily due to a disturbance in cerebral cortical function and that median SEP may be used for early detection of HE and for monitoring its clinical course. In addition, absence of the N20-P25 component, or presence of only the N20 component of the wave complex in fulminant hepatic failure is associated with high mortality, whereas presence of late cortical components in HE is usually associated with reversibility of clinical course. Central conduction time (CCT) of the BAEP is prolonged in patients with WD, alcoholic liver disease and liver cirrhosis due to hepatitis B. Furthermore, BAEP abnormality is most severe in WD, followed by alcoholic liver disease, and finally hepatitis B. Peripheral nerve conduction as determined by the N9 latency of SEP is slowed in alcoholic liver disease and liver cirrhosis of chronic hepatitis B, but normal in WD. Our studies, therefore, suggest that evoked potentials may be useful in the evaluation of both CNS and PNS functions in various liver diseases and also in the diagnosis and monitoring of HE. PMID- 9407350 TI - Epidemiological characteristics and risk factors of hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is one of the major cancers in the world. There is a striking variation in HCC incidence rates between various countries, with a highest-to-lowest ratio of 112.5 for males and 54.7 for females. The high-risk populations are clustered in sub-Saharan Africa and eastern Asia. The male-to female ratio for HCC ranges from < 1 to 6.4 and mostly from 2 to 4. There exist significant variations in the incidence of HCC among different ethnic groups living in the same area and among migrants of the same ethnic groups living in different areas. The age curves of HCC are significantly different in various countries, suggesting variability in exposure to risk factors. Chronic carriers of hepatitis B and C viruses (HBV and HCV, respectively) have an increased risk of HCC. The relative and attributable HCC risk of HBV and HCV carrier status varies in different countries. There exists a synergistic interaction on HCC between the two viruses. Aflatoxin exposure, cigarette smoking, heavy alcohol consumption, low vegetable intake, inorganic arsenic ingestion, radioactive thorium dioxide exposure, iron overload and the use of oral contraceptives and anabolic steroids have been documented as HCC risk factors. Recent molecular epidemiological studies have shown that low serum retinol levels as well as elevated serum levels of testosterone, neu oncoprotein and aflatoxin B1-albumin adduct are associated with an increased HCC risk. There is a synergistic interaction on HCC between chronic HBV infection and aflatoxin exposure. Familial aggregation of HCC exists and a major susceptibility gene of HCC has been hypothesized. Patients of some genetic diseases are at an increased risk of HCC. The genetic polymorphisms of cytochrome P450 2E1 and 2D6 and arylamine N acetyltransferase 2 are associated with the development of HCC. A dose-response relationship between aflatoxin exposure and HCC has been observed among chronic HBV carriers who have null genotypes of glutathione S-transferase M1 or T1, but not among those who have non-null genotypes. Human hepatocarcinogenesis is a multistage process with the involvement of a multifactorial aetiology. Gene environment interactions are involved in the development of HCC in humans. PMID- 9407351 TI - Molecular mechanism of hepatocarcinogenesis. AB - To clarify the relative role of hepatitis C virus (HCV) and hepatitis B virus (HBV) in hepatocarcinogenesis in hepatitis B surface antigen (HBsAg)-negative hepatocellular carcinoma (HCC) in Taiwan, polymerase chain reaction (PCR) was used to detect the HCV-RNA and HBV-DNA sequences in the serum and liver tissues from 31 HBsAg-negative HCC patients. Twenty-one were positive for antibody to HCV (group 1) and 10 were negative (group 2). Hepatitis C virus-RNA was detected by PCR in the serum of 16 group 1 patients and in the liver tissue of 17; while HBV DNA was found in the liver tissue of only four, and no HBV-DNA was found in the serum. Hepatitis C virus RNA was detected in the serum of one group 2 patient and in the liver tissue of another. In contrast, HBV viral DNA was found in the serum of four group 2 patients and in the liver tissues of five patients. This indicates that HCV plays an important role in hepatocarcinogenesis in HBsAg negative patients in Taiwan, especially in those with antibody to HCV. In those without antibody to HCV, HBV might still be associated with the development of HCC in a significant proportion of such patients. In order to study the role of the p53 mutation in hepatocarcinogenesis, we investigated the status of the p53 mutation in 61 HCC samples from Taiwan. The exon 5 to 8 of the p53 gene in the tumour tissue of 61 HCC were amplified and sequenced. A total of 20 cases (32.8%) were found to have mutations: 36.6% (15/41) from the HBsAg-positive group and 25.0% (5/20) from the HBsAg-negative group. The corresponding normal liver showed no mutation. The mutation is widely distributed throughout the exon 5 to 8. Only four cases (6.6%), all positive for HBsAg, had a specific hotspot mutation at codon 249 with G to T transversion. These results show that scattered point mutations in p53 are not uncommon in HCC samples from Taiwan and may be important in the development of this cancer. However, the aflatoxin-related specific mutation seems much less related to the genesis of HCC in Taiwan. To study the role of telomerase activity in hepatocarcinogenesis, a total of 39 HCC tissues and the corresponding non-tumour liver tissues were analysed. The results showed that telomerase activity was detected in all the 39 tumour tissues, while it could be detected in six of the 39 non-tumour liver tissues. The high positive rate of telomerase activity in HCC samples suggests that telomerase activity is closely related to the development or progression of HCC. To determine whether exon 1 and exon 2 of the p16 gene are altered in HCC, thirty-four tumours from 30 HCC patients were examined by DNA sequencing analysis of PCR-amplified genomic DNA. Homozygous deletions of MTS1/p16/CDKN2 exon 1 were identified in 1/34 primary tumours (3%), no mutations or rearrangements were found in these specimens. These data suggest that alterations of MTS1/p16/CDKN2 gene are rarely found in HCC, and might play little role in the development of this cancer. To study the clonality of HCC, 18 patients with multiple HCC, most of them small in size, were analysed by DNA fingerprinting. In patients positive for hepatitis B surface antigen, the integration pattern of hepatitis B viral DNA in liver tissue was also analysed. The results by both methods showed that 8/9 hepatitis B surface antigen-positive patients were different in clonality. In the remaining nine patients negative for hepatitis B surface antigen, four had different band patterns in their tumours by DNA fingerprinting. This study indicated that polyclonality of multiple HCC was rather frequent and it highlighted the importance of eliminating the underlying cause of liver injury to improve the survival of these patients. Microsatellite markers were used to study the genetic changes of HCC. Thirty cases of HCC, most of them small in size, were studied. A total of 242 microsatellite markers mapping to 1-22 and X chromosomes was used. The results showed that the range of loss of het PMID- 9407352 TI - Hepatocellular carcinoma: clinicopathological aspects. AB - The histopathology and clinical picture of hepatocellular carcinoma (HCC) varies between individual patients and regions. These variations are perhaps due to differences in the genetic alterations that precede hepatocarcinogenesis. In this study, the clinicopathological features of HCC were compared between southern African blacks and Japanese, indicating large differences in the frequency of underlying cirrhosis, grade of cancer cell differentiation and clinical course. Intra-abdominal bleeding and febrile, rapidly progressive HCC are more common among blacks. Such a difference is accounted for, in part, by frequent encapsulation of the tumour which is well differentiated, and grows slowly in an expanding fashion in Japan. Encapsulated HCC was not seen among the black patients studied. Other distinct clinicopathological types discussed in this paper include diffuse-type HCC which is usually caused by multiple portal spread occurring almost simultaneously; the clinical course is fulminant. Sclerosing carcinoma is frequently associated with hypercalcaemia in the United States, but not in Japan. Fibrolamellar carcinoma is nearly non-existent in Asia, whereas it is common among young adults in the West. Its prognosis is generally better than ordinary HCC. Hepatocellular carcinoma has a strong propensity to invade vessel and duct systems. Portal invasion does not produce distinct clinical signs although it may aggravate portal hypertension. Patients with tumour occlusion in the major portal vein may give rise to ischaemic hepatitis when blood pressure drops suddenly in the preterminal stage. Liver parenchyma develops submassive necrosis and clinically there is an acute rise in alanine aminotransferase (ALT). Invasion into a major hepatic vein and the inferior vena cava also occurs, but less frequently compared with portal invasion. The patient can live even with a tumour thrombus in the atrium crossing the tricuspid valves. Intraductal invasion causes acute jaundice as well as an occasional haemobilia with pain. We recently found that a distinct pathological type called 'extrahepatic growth' or 'pedunculated HCC' develops as a result of fusion of right-sided adrenal metastasis of HCC and the liver, perhaps through the 'adreno-hepatic fusion' which is rather common in cirrhotic livers. PMID- 9407353 TI - Non-surgical treatment of hepatocellular carcinoma. AB - A decade ago, surgery was the only satisfactory treatment modality for hepatocellular carcinoma (HCC), but it was limited only to selected cases. For the majority of cases of HCC, systemic chemotherapy was one of the few treatment alternatives, but provided only marginal benefit. In the past 20 years, diagnostic methods have improved to an extent that small HCC less than 1 cm can be detected. Moreover, non-surgical treatment is available, of which regional therapy has been shown to prolong patients' survival, and may even replace surgical resection in some cases. Regional therapy is indicated for the treatment of HCC when there is no extrahepatic metastasis and the patient has adequate liver function reserve, thus permitting repeated therapy. Transcatheter hepatic arterial embolization (TAE) using various embolizers has been well documented to include controlled studies. However, it is not indicated for patients with thrombosed main portal veins. Its therapeutic effect is also doubtful when the tumour is infiltrative in nature or is hypovascular, too large or too small. Additional chemotherapeutic agents mixed into the embolizer with lipiodol and degraded starch microspheres or styrene-maleic acid-neocarzinostatin in which chemotherapeutic agents are embedded, are used with a better response, but the survival rate has not shown significant improvement. Ultrasound-guided local injection therapy is another new method of treatment of HCC. Of these techniques, percutaneous ethanol injection therapy (PEIT) is widely used with excellent results for small, encapsulated tumours in livers with less than three HCC. Percutaneous ethanol injection therapy can also be used in cases with portal vein thrombosis, but it is not suitable for patients having coagulopathy or ascites. Using acetic acid, OK-432, interferon or anti-cancer drugs in the injection therapy shows no further benefit over ethanol alone. Transcatheter echoguided thermotherapy or cryotherapy has been reported in small series of patients, as has target therapy with immune or radiotherapy and conformal radiotherapy. Preliminary studies show encouraging results. Systemic therapy with either single drug or multidrugs is ineffective, with a response rate of less than 20%. Immunotherapy, such as with interferon or other cytokines, is not beneficial. Hormone therapy has not been promising, except for treatment with tamoxifen, which has been reported to show some beneficial effect. Gene therapy is still in its infancy. In summary, recent progress in non-surgical treatment of HCC has resulted in a breakthrough of regional therapy looking quite promising. Moreover, a combination of different types of regional therapies may yield better outcomes in selected individuals. PMID- 9407354 TI - Partial hepatic resection for hepatocellular carcinoma. AB - This review summarizes the efficacy of the most common therapeutic option for hepatocellular carcinoma (HCC), partial hepatic resection, taking into account not only its antitumoural effect, but also its consequences on survival. Partial hepatic resection results in 5 year survival rates as high as 45% in more favourable subgroups having: small tumours, well-differentiated tumours, unifocal tumours, a lack of vascular invasions, an absence of cirrhosis, and the fibrolamellar variant. Resection has been limited primarily by low resectability rates and recurrent disease. However, surgical resection in the form of partial hepatectomy is the preferred treatment for HCC. The early detection of tumours by screening high-risk populations is crucial. During the 12 year period between 1983 and 1994, hepatic resections were carried out in 382 patients with HCC. One hundred and fifty-three (40%) had HCC smaller than 5 cm in diameter. There were 294 male and 88 female patients, with an average age of 52.3 years. Among them, 45% had liver cirrhosis and 73% were positive for hepatitis B surface antigen. Two hundred and eighteen (57%) were positive for hepatitis C virus circulating antibodies (since 1991). Operative mortality was 3.9%. The overall survival rates at 1, 3 and 5 years were 71, 52 and 46%, respectively. Sex, cirrhosis, Child's staging, surgical procedure, blood loss, pathological pattern, presence of capsule, surgical margin and DNA ploidy appeared to be factors not related to prognosis. However, alpha-fetoprotein level, size (whether less than or greater than 5 cm), and vascular invasion were factors which significantly affect survival. PMID- 9407355 TI - Hepatitis viruses and liver transplantation. AB - Acute and chronic liver diseases related to hepatitis viruses are the main indications for liver transplantation. The risk of viral reinfection after transplantation is the main limiting factor in these indications. The risk of viral B reinfection is: 80% in the absence of prophylaxis; is related to the presence of active viral B replication prior to transplantation; is higher in patients with chronic liver disease, rather than with fulminant hepatitis; and is higher in patients with hepatitis B virus (HBV)-related liver disease alone rather than in those with HBV-hepatitis delta virus (HDV) infection. Post transplant long-term passive antibody to hepatitis B (anti-HB) immunoprophylaxis reduces the risk of HBV recurrence to 30% in patients with HBV cirrhosis, and to less than 10% in those with fulminant hepatitis B. Patients with HBV-HDV liver disease receiving passive anti-HB immunoprophylaxis are at low risk of HBV recurrence (10-15%), but at high risk of HDV recurrence (80%). However, HDV reinfection of the graft has no clinicopathological consequence in the absence of concomitant HBV reinfection. The five year survival of patients transplanted for HBV cirrhosis and for HDV cirrhosis at the Hepatobiliary Center, Hopital Paul Brousse is 72% and 85%, respectively. Hepatitis B virus reinfection of the graft is characterized by a high level of viral replication, and a chronic outcome. Antiviral treatments such as ganciclovir, adenine arabinoside monophosphate, famcyclovir, and lamivudine have a place after transplantation and may stop HBV replication, ganciclovir, famcyclovir and lamivudine should be continued for several months and in some cases indefinitely. Hepatitis C virus reinfection is almost constant, assessed by the persistence of hepatitis C virus (HCV)-RNA in the serum in 90% of cases. Acute lobular hepatitis appeared in 75% of patients at a median of 4 months post transplantation with a range of between 23 days and 4 years. In our series, the 5 year actuarial rate of HCV acute hepatitis on the graft, chronic hepatitis, and cirrhosis, is 75, 60, and 8%, respectively. Hepatitis C virus RNA level is dramatically increased after transplantation and seems to correlate with the occurrence of acute hepatitis on the graft. A positive relation between genotype 1b and prevalence and severity of HCV hepatitis on the graft have been suggested in European series. There is no demonstrated way to prevent HCV reinfection. The use of interferon for the treatment of HCV hepatitis on the graft was disappointing due to a poor antiviral effect and the occurrence of chronic rejection episodes in some patients. Promising results of the combination of interferon and ribavirine have been reported and need confirmation. The 5 year survival of patients transplanted for viral C cirrhosis at the Hepatobiliary Center, Hopital Paul Brousse is 78%. In conclusion, patients with HBV cirrhosis and without HBV replication are candidates for liver transplantation. Long-term passive anti-HB prophylaxis is the best way to prevent HBV recurrence. Patients with HBV replication should be included in protocols using combinations of antiviral treatments and passive anti HB immunoprophylaxis. Viral C reinfection is frequent, but medium-term survival is good. However, long-term graft and patient survival remains unknown and methods to prevent and treat HCV reinfection on the graft are needed. PMID- 9407356 TI - Living related donor liver transplantation. AB - Living related liver transplantation (LRLT) has been developed in response to the paediatric organ donor shortage. According to the International Living Donor Registry, 521 transplants had been performed in 515 patients between December 8 1988 and January 19 1996 in 30 centres worldwide. The overall actuarial patient and graft survival rates were 82.7 and 80%, respectively. Between June 17 1994 and November 30 1996, the authors performed 11 LRLT at the Chung Gung Memorial Hospital. The living donors consisted of 10 mothers and one father. The mean graft weight was 303 g and the mean graft recipient weight ratio was 2.2%. Donor hepatectomy was performed without vascular inflow occlusion. The intra-operative blood loss ranged from 30 mL to 120 mL with an average of 61 mL, and blood transfusion was not required in all donors both intra-operatively and during the postoperative period. Underlying diseases of the recipients were biliary atresia (n = 10) and glycogen storage disease (n = 1). The mean graft cold ischaemia time was 106 min, the mean second warm ischaemia time was 51 min and the mean interval between portal and arterial reperfusion was 81 min. The initial LRLT results were promising with all donors having been discharged without complication. The recipients experienced a few complications, all of which were manageable with early intervention. All 11 recipients are alive and well. These are encouraging results and the authors hope to expand the use of live donors for liver transplantation to cope with demand. PMID- 9407357 TI - Current therapeutic trends in therapy for chronic viral hepatitis. AB - Hepatitis B virus (HBV), hepatitis C virus (HCV) and hepatitis delta virus (HDV) are associated with clinically significant chronic infection that may lead to the development of cirrhosis or even hepatocellular carcinoma (HCC). Intervention at the earliest possible stage is needed to prevent such untoward sequelae. Currently, interferon (IFN) is the only approved and widely used agent for the treatment of these infections, including in HBV patients with precore mutant hepatitis or decompensated cirrhosis, but its efficacy is far from satisfactory. Corticosteroid priming has been shown to increase the efficacy of IFN therapy in HBV patients with low abnormal serum transaminase levels, but only a few responders will clear serum hepatitis Bs antigen (HBsAg). Ongoing randomized controlled trials of thymosin alpha 1, lamivudine and famcyclovir have demonstrated encouraging preliminary results. Therapeutic vaccines, such as polypeptides with human leucocyte antigen (HLA)-specific hepatitis B core antigen (HBcAg) epitopes, are under phase II/III clinical trial. For HDV infection, the use of IFN in the early phase of acute superinfection tends to prevent chronic progression. For HCV infection, IFN used at higher doses for a longer period of time is associated with a higher sustained response, but overall it is still not satisfactory. The combined use of ribavirin or corticosteroid priming may improve the effect of IFN therapy by enhancing the durability of the response. Interferon in the acute phase of HCV infection may also prevent chronic progression. There is evidence to suggest that IFN therapy, when associated with response, tends to reduce the risk of cirrhosis or HCC and prolongs survival. There is no doubt that satisfactory treatment of chronic viral infection will require more effective agents and demand optimal treatment strategies, many of which are yet to be found. PMID- 9407359 TI - Prevention of hepatitis B and C virus infection for prevention of cirrhosis and hepatocellular carcinoma. AB - In Taiwan, cirrhosis and hepatocellular carcinoma (HCC) have been common in medical practice since the 1960s. In 1969, Taiwan was shown to be a hyperendemic area of hepatitis B virus (HBV) infection with a high rate of hepatitis B surface antigen (HBsAg) positivity, 19% of the population being infected before the fourth decade of life. There is evidence indicating that more than 80% of chronic hepatitis, cirrhosis and HCC are the sequelae of chronic HBV infection. In 1984, after 3 years of preparation, a programme to control cirrhosis and HCC began. All neonates born to HBsAg+ mothers were given Pasteur plasma-derived vaccine 5 micrograms i.m. at 1, 5 and 9 weeks with a booster at 12 months. In 1986, all neonates were included in this programme. In addition, beginning in 1987, all non vaccinated preschool children were also immunized and susceptible medical personnel and people from HBsAg+ households were recommended to receive the vaccine. Using data obtained from the 7-year evaluation study on the efficacy of this vaccine and some historical data, the HBsAg positivity rate in people born in the first few years after 1986 was estimated to be 2.6%. This rate is expected to decrease to 0.2% in those born after around 1990. In July 1992, an anti hepatitis C virus (HCV) test was included in blood donor screening tests. This was followed by a decrease in the incidence of post-transfusion hepatitis (PTH) from 13 to 2.5% and there have been no anti-HCV+ PTH cases since. However, without immunization, the prevalence of HBsAg decreased among children in Taipei in 1989. This coincided with the widespread use of disposable syringes and needles and an improvement in the sterilization of medical instruments. Therefore, it is likely that HCV infection may also decrease as a result of these practices. Through the use of immunization and improved medical procedures, chronic hepatitis, cirrhosis and HCC may decrease in Taiwan by around 95%. PMID- 9407358 TI - Nucleic acid-based antiviral and gene therapy of chronic hepatitis B infection. AB - Persistent hepatitis B virus (HBV) infection often leads to the development of chronic hepatitis, cirrhosis and hepatocellular carcinoma. There is a need to develop new antiviral approaches for the treatment of this disease. We have explored various nucleic acid-based strategies designed to inhibit HBV replication including: the use of antisense RNA and DNA constructs, DNA-based immunization techniques to stimulate broad-based cellular immune responses with particular emphasis on the generation of cytotoxic lymphocyte (CTL) activity to viral structural proteins, hammerhead ribozymes to cleave HBV pregenomic RNA in vitro and dominant negative HBV core mutant proteins as inhibitors of nucleocapsid formation within cells. In order to optimize these antiviral effects, various novel expression vectors have been developed to deliver such DNA constructs to cells. For example, adenoviral vectors carrying genes that encode for dominant negative proteins have been employed to transfect hepatocytes in vitro and in vivo. In addition, plasmid vectors have been produced to promote expression of HBV structural genes following injection into muscle cells as a means to stimulate the host's cellular and humoral immune response in the context of histocompatibility antigen (HLA) class I and II antigen presentation. These experimental approaches may have important implications for the generation of efficient antiviral effects during chronic HBV infection. PMID- 9407360 TI - Visual function measures in experimental glaucoma. PMID- 9407361 TI - Cystic epithelial ingrowth after goniotomy for congenital glaucoma. A clinicopathologic report. AB - PURPOSE: To describe the clinical and histopathologic findings of a child with congenital glaucoma who developed cystic epithelial ingrowth after goniotomy. METHODS: A five-month-old boy was diagnosed with congenital glaucoma due to Axenfeld syndrome. Goniotomy was performed immediately after the clinical diagnosis in his left eye. Cystic epithelial ingrowth was noted two years postoperatively. The cyst was incompletely excised elsewhere, and recurred two years later. RESULTS: Cystic epithelial ingrowth expanded from the 2 o'clock to the 4 o'clock position involving the chamber angle. Best corrected preoperative visual acuity was 20/600 in his left eye. Cystic epithelial ingrowth was treated by block excision (8.0 mm) and tectonic corneoscleral grafting (8.1 mm). Best corrected postoperative visual acuity was 20/200 and the intraocular pressure was 8 mmHg without antiglaucomatous medication. There was no recurrence of the cyst during the 31 months of follow-up. Histopathologic results found that cystic epithelial ingrowth consisted of nonkeratinizing squamous epithelium with goblet cells covering Descemet's membrane, the chamber angle structures, the anterior face of the ciliary body, and the iris. CONCLUSIONS: Cystic epithelial ingrowth is a rare complication after goniotomy. Block excision with tectonic corneoscleral grafting is the treatment of choice for cystic and diffuse sheet like epithelial ingrowth. PMID- 9407362 TI - Corneal melting after avulsion of a Molteno shunt plate. AB - PURPOSE: We report a case of Molteno shunt avulsion after blunt trauma resulting in corneal melting. The implant's connecting tube had been placed above the superior rectus muscle. METHODS: Case report. RESULTS: After blunt trauma, the secondary plate of a double-plated Molteno implant became completely dislodged anteriorly onto the patient's cornea. The corneal tissue entrapped beneath the avulsed plate was found to have extensive melting requiring emergency explantation of the Molteno implant and corneal grafting. CONCLUSIONS: Corneal melting after Molteno shunt avulsion is a novel finding. Placing a double-plated Molteno implant's connecting tube under the superior rectus muscle might decrease the risk of shunt avulsion after trauma. PMID- 9407364 TI - A comparison of healthy, ocular hypertensive, and glaucomatous optic disc topographic parameters. AB - PURPOSE: To compare the optic discs of 62 healthy individuals 68 patients who have ocular hypertension (OH), and 182 patients with primary open-angle glaucoma (132 high-tension glaucoma (HTG) and 50 normal-tension glaucoma (NTG)), and determine whether disc size exerted an influence on the group differentiation. PATIENTS AND METHODS: Standard criteria were used to define glaucoma and normality. Ocular hypertension was defined as having raised intraocular pressure, a normal visual field, and a healthy optic disc/retinal nerve fiber layer (RNFL). The optic disc of one eye from each individual was analyzed using a confocal scanning laser ophthalmoscope (Heidelberg Retina Tomograph software version 1.11, Heidelberg Engineering, Heidelberg, Germany). Thirteen topographical, volumetric, and shape parameters were compared between the three diagnostic groups. In addition, the individuals were divided into subgroups on the basis of disc size to determine any effect of disc size on the differentiating ability of the confocal scanning laser ophthalmoscope. Differences between the groups were evaluated using an analysis of variance. RESULTS: Glaucomatous optic discs were found to differ from both healthy and OH discs, although no differences in disc area between the groups were identified. On the basis of disc size, differentiating the glaucomatous discs was best for midsized discs of 2 mm2 to 3 mm2. However, no difference was found between healthy and OH discs, even when allowing for disc size. CONCLUSIONS: Ocular hypertensive optic discs (with a clinically normal appearance) could not be distinguished from healthy discs using a confocal scanning laser ophthalmoscopic technique. Glaucomatous optic discs were found to differ from both healthy and OH discs, with a limited effect of disc size. PMID- 9407363 TI - Scanning laser polarimetry in corneal haze after excimer laser refractive surgery. AB - PURPOSE: We evaluate the technique of retinal nerve fiber layer thickness measurement using scanning laser polarimetry in cases of central corneal haze due to photorefractive keratectomy. METHODS: Nerve fiber layer thickness was measured using a Nerve Fiber Analyzer II (Laser Diagnostic Technologies, Inc., San Diego, CA, U.S.A.) in eight eyes of eight individuals with mild to pronounced corneal haze owing to earlier excimer laser refractive surgery, and in nine eyes of nine control volunteers. RESULTS: Neither total and sectorial nerve fiber layer thickness values, nor superior/ inferior quadrant thickness ratio differed in a statistically significant manner between the two groups (unpaired t test, p > 0.05). Reproducibility in the "haze" and control groups was 4.3% and 3.4%, respectively, for mean nerve fiber layer thickness calculated from all quadrants around the disc. The figures were 4.9% and 5.2%, respectively, for the superior quadrant thickness; 8.2% and 8.8%, respectively, for the temporal thickness values; 5.4% and 4.6%, respectively, for the inferior thickness values; and 4.1% and 5.0%, respectively, for the nasal quadrant thickness values. For the superior/inferior quadrant thickness ratio the reproducibility was 4.4% and 6.8%, respectively. None of the corresponding values for reproducibility differed significantly between the two groups (F-test, p > 0.05). CONCLUSIONS: Corneal haze did not cause an artificial increase of the thickness values measured with scanning laser polarimetry and did not diminish the reproducibility of the measurement. The results suggest that scanning laser polarimetry may be a suitable method for the precise measurement of the nerve fiber layer thickness even in eyes with persistent corneal haze after excimer laser refractive surgery. PMID- 9407365 TI - Sector-based analysis of optic nerve head shape parameters and visual field indices in healthy and glaucomatous eyes. AB - PURPOSE: To evaluate the correlations between the sector optic nerve head parameters measured by Heidelberg Retina Tomograph (HRT, Heidelberg Engineering, Heidelberg, Germany), version 1.11S, and the visual field. METHODS: One eye was randomly chosen from 55 individuals with glaucoma and 50 healthy individuals. Each participant had at last one Humphrey visual field, program 30-2 (Humphrey Instruments, San Leandro, CA, U.S.A.), and three 10 degrees HRT pictures. From the mean of the three HRT pictures, global measurements, superior (45 degrees-135 degrees), nasal (135 degrees-225 degrees), inferior (225 degrees-315 degrees), and temporal (315 degrees-45 degrees) sector measurements were calculated for the following parameters: disc area, effective area, area below reference, mean height of contour, volume below surface, volume above surface, volume below reference, volume above reference, and third moment. From the visual field results, mean deviation (MD), superior MD, and inferior MD were calculated. For each HRT parameter we calculated the "r" Pearson correlation with the corresponding visual field measures. RESULTS: Within the combined healthy and glaucomatous groups we found highly significant (p < 0.001) correlations between the following HRT parameters and the visual field MD: inferior and mean high of contour (r = -0.53), inferior and third moment (r = -0.52), global and third moment (r = -0.49), inferior and volume above reference (r = 0.47), superior and third moment (r = -0.46), and superior and area below reference (r = -0.44). Correlations between global mean deviation and nasal or temporal sector parameters were generally smaller and less significant. CONCLUSIONS: Inferior and superior HRT sector parameters were correlated with the respective visual field indices. In many cases these correlations were as strong or stronger than with the global equivalent shape measures. PMID- 9407366 TI - Physicians' guide to interactions between glaucoma and systemic medications. AB - PURPOSE: To develop a Physicians' Guide to assist clinicians in the concomitant use of glaucoma and systemic medications. METHODS: The records of 100 consecutive patients with chronic open-angle glaucoma were retrospectively studied to determine the most common systemic medications that are prescribed in this population. The ten most common drug classes were then used to construct a guide to potential interactions and side effects when these medications are used concomitantly with glaucoma drugs. RESULTS: Eighty-four patients were receiving 1 or more medications (a mean of 3.5) for a mean of 2.6 systemic conditions. Systemic antihypertensive agents was the most common class of drugs, being used by 48 patients. Aspirin, the most common single systemic drug, was being used by 25 patients. CONCLUSIONS: A high percentage of patients with chronic open-angle glaucoma receive a wide variety of medications for coexisting systemic disorders. The concomitant use of glaucoma and systemic medications creates the potential for drug interactions, as well as side effects for both groups of drugs, for which the Physicians' Guide may be beneficial. PMID- 9407367 TI - Reproducibility of retardation measurements with the nerve fiber analyzer II. AB - PURPOSE: To evaluate the reproducibility of a scanning laser polarimeter, the Nerve Fiber Analyzer II (NFA II, Laser Diagnostic Technologies, San Diego, CA, U.S.A.). METHODS: Five independent retardation maps of the peripapillary retina of five normal eyes were acquired by three experienced operators (including V.S.G.) on each of three separate days for a total of 45 retardation maps per patient. Two methods of image processing, one using a baseline image and another using the individual scans, were used to compare the reproducibility of three summary measures, average retardation, integral, and retardation ratio. RESULTS: The average standard deviation (and its 95% confidence interval) of average retardation within a 10-pixel-width-band of the 9 baseline images was 0.43 degree (0.36-0.51 degree) with a mean coefficient of variation of 4.2% (3.8-4.5%). In a random effects model, each of the three retinal nerve fiber layer (RNFL) summary measures varied significantly by patient (p < 0.016), but not by operator (p > 0.19), or operator by patient interaction (p > 0.524). In addition, there was small, but statistically significant day-by-operator-within-patient (intraobserver) variation in the random effects model. CONCLUSIONS: These results suggest that the NFA II provides reproducible measurements and that, on average, measurements obtained by separate operators on different days are similar. PMID- 9407368 TI - Experimental glaucoma: perimetric field defects and intraocular pressure. AB - PURPOSE: To investigate the relationship between intraocular pressure (IOP) and the progression of visual field defects caused by experimental glaucoma in Macaca mulatta monkeys. METHODS: Perimetric fields were measured by behavioral methods in 18 rhesus monkeys during the course of unilateral glaucoma produced by argon laser treatment of the trabecular meshwork. The monkeys' IOPs were measured by applanation tonometry. Visual field defects were quantified by the mean deviation perimetric index from Humphrey Field Analyzer C24-2, model 630 (Humphrey Allergan, San Leandro, CA, U.S.A.), full-threshold data. RESULTS: The monkeys' eyes demonstrated considerable variability in their susceptibility to pressure induced neural damage. For 10 of the monkeys, significant field defects were correlated with the increases in their IOPs and the defects progressed monotonically to end-state glaucoma. For the other monkeys, the mean deviation index was not well correlated with IOP; some eyes withstood pressures in excess of 35 mm Hg for several months before significant reduction in visual sensitivity. However, once they began, the rate of progression of field defects was similar across subjects. CONCLUSIONS: Laser ablation of the trabecular meshwork in monkeys provides a model for investigations of the effects of IOP that are not confounded by other ocular or visual disorders. Behavioral perimetry showed the same intersubject variability in the effects of elevated IOP on visual field sensitivities of monkeys that are common with high-tension glaucoma or ocular hypertension in patients. Thus, these investigations provide additional support for the use of the model for a wide variety of clinical investigations on glaucoma. PMID- 9407369 TI - Ascorbate stimulates type I and type III collagen in human Tenon's fibroblasts. AB - PURPOSE: To better understand wound healing after glaucoma filtration surgery by measuring the production of type I and type III collagen in cultured Tenon's fibroblasts and determine the effect of ascorbic acid on collagen subtype production. METHODS: An ELISA-type dot blot assay was used to directly measure the production of types I and III collagen by subconfluent cultures of fibroblasts from human Tenon's capsule. Because ascorbic acid is both high in aqueous humor and necessary for the production of collagen, we measured the dose response of type I and type III collagen production to ascorbic acid. RESULTS: Ascorbic acid stimulated an increase in collagen production that reached a maximum level at 100 micrograms/ml. This is approximately half of the ascorbic acid concentration found in human aqueous humor. Unlike previous reports, we found no toxic effects from ascorbic acid at concentrations as high as 250 micrograms/ml over a 24-hour period. The lack of toxicity may result from the use of serum-free media in the assay. CONCLUSIONS: This culture system will be useful for exploring factors that may alter collagen production and could potentially affect wound healing. PMID- 9407370 TI - Treatment of progressive normal-tension glaucoma. PMID- 9407371 TI - Factors influencing blood flow in the optic nerve head. PMID- 9407372 TI - Diode laser transscleral cyclophotocoagulation. PMID- 9407373 TI - Iris-color change developed after topical isopropyl unoprostone treatment. AB - PURPOSE: To present a case in which iris pigmentation developed after treatment with isopropyl unoprostone, an analogue of a prostaglandin metabolite. METHODS: Case report. RESULTS: A Japanese man with dark brown irises, treated unilaterally with isopropyl unoprostone, developed iris-color change in the treated eye after a 20-month treatment. CONCLUSIONS: Isopropyl unoprostone can induce iris pigmentation as does latanoprost. PMID- 9407374 TI - Early successful treatment of postoperative necrotizing pseudomonas scleritis after trabeculectomy. AB - PURPOSE: The authors describe a patient who developed pseudomonas scleritis after a routine trabeculectomy. METHOD: The patient underwent trabeculectomy for poorly controlled intraocular pressure and progressive visual field loss. On the second postoperative date he developed severe pain, significant anterior chamber reaction, and hypotony. Scleral cultures taken at the time of surgical choroidal drainage grew pseudomonas aeruginosa. RESULT: Surgical reconstruction of the necrotic scleral area and intensive antibiotic treatment lead to a successful outcome. CONCLUSION: Early recognition and aggressive treatment with antibiotics initially, followed by surgical debridement of necrotic tissue, resulted in an unexpected successful outcome in a patient with pseudomonas scleritis. PMID- 9407375 TI - Optic disk. PMID- 9407376 TI - Blockage of a Krupin valve tube. PMID- 9407377 TI - Increase in neutralizing antibody titer against sequential autologous HIV-1 isolates after 16 weeks saquinavir (Invirase) treatment. AB - The humoral immune response to HIV infection plays an important role in determining disease progression. Few and discordant results correlate changes in neutralizing antibody (NtAb) titer with antiretroviral treatment. The NtAb titer against autologous-HIV was evaluated in 33 patients treated with the protease inhibitor saquinavir (SQV, Invirase) and zidovudine (ZDV) alone or in combination. Ten out of 33 (30%) patients showed a significant increase (4-fold or greater) in NtAb titer from baseline in response to the initiation of therapy. A significant correlation (P = 0.007) was found between an increase in NtAb titer and treatment with SQV alone (5 subjects) or in combination (5 subjects). A significant decrease in NtAb titer was detected in 7 patients, 5 of whom were treated with ZDV alone. After one year of therapy a significant decrease in HIV RNA copy number (> 0.5 log) with respect to baseline value was detected only in patients treated with SQV alone or in combination. Patients with increased NtAb titer showed a significantly reduced HIV-RNA copy number and increased CD4+ cell count at week 16 of treatment which were sustained up to week 52. These data suggest that treatment with SQV can improve neutralizing activity against autologous virus as well as bring about a significant and sustained reduction in viral load. PMID- 9407378 TI - Detection of herpes simplex virus type-specific antibodies by an enzyme-linked immunosorbent assay based on glycoprotein G. AB - In order to develop a simple and quantitative method to detect herpes simplex virus (HSV) type-specific antibodies, the usefulness of an enzyme-linked immunosorbent assay (ELISA) using HSV glycoprotein G (gG) captured on a plate by monoclonal antibodies as antigen was studied. The gG1- and gG2-specific IgG antibody activities were measured by the ELISA for 54 sera which had been collected from culture-proven genital herpes patients and pre-characterized by an immunodot assay using purified gG antigens. Thirty control sera without antibodies against the HSV whole antigens were also included. In comparison with the immunodot assay as standard, the sensitivities of the ELISA were 88.9% (32/36) for HSV-1 antibody and 89.2% (33/37) for HSV-2 antibody and the specificities were both 100%. Sera taken within a few months after primary infection tended to give false negative results. The HSV type-specific ELISA based on easy-to-prepare gG antigens might be useful to help improve the serological assessment of HSV infections. PMID- 9407379 TI - The detection of intrathecal synthesis of anti-herpes simplex IgG antibodies: comparison between an antigen-mediated immunoblotting technique and antibody index calculations. European Union Concerted Action on Virus Meningitis and Encephalitis. AB - The detection of intrathecal antibody synthesis was compared by the calculation of antibody indices (AI) derived from ELISA techniques with the detection of virus-specific oligoclonal IgGs by an antigen-mediated capillary blot technique. Twenty-seven paired serum and cerebrospinal fluid (CSF) samples were examined from 15 immunocompetent patients with herpes simplex virus encephalitis (HSE) diagnosed by PCR on early CSF samples. These techniques were also applied to paired samples from 20 multiple sclerosis (MS) patients, 10 patients with other inflammatory neurological diseases and 10 patients with non inflammatory neurological disorders. There was a good correlation between the results obtained by AI and those obtained by immunoblotting, especially in HSE (2 discordant results out of 27). Discrepancies were more frequent (25%) in MS patients where a "polyspecific" reaction characterized by low affinity antibodies is known to occur. Some of the discrepancies could, in part, be due to serological cross reaction with varicella zoster virus. PMID- 9407380 TI - Cloning, expression, and immunogenicity of the assembly protein of varicella zoster virus and detection of the products of open reading frame 33. AB - Herpesviruses produce assembly proteins (AP) that act as scaffolding proteins for the assembly of the viral capsids. The products of the assemblin gene, which encodes both maturational protease and AP, have been established for herpes simplex virus type 1 (HSV-1) and human cytomegalovirus (CMV). We cloned an inframe ORF (encoding amino acids 304-605), found within the ORF 33 assemblin gene of VZV, into a yeast expression vector. The 34-kDa AP was expressed as a fusion protein with the particle-forming Ty p1 protein, resulting in high-level production of hybrid AP-virus-like particles (AP-VLPs). When AP-VLPs were injected into mice and rabbits, antibodies were produced that reacted with, but that did not neutralise, native VZV. Three of four inbred strains of mice immunised with AP-VLPs produced a VZV-specific T-cell response. The mouse and rabbit sera reacted with six bands on native VZV by Western blot analysis. The dominant bands were found at 34 and 38 kDa. Bands were also seen at 66, 63, 41, and 31 kDa. The 38-kDa protein may represent the mature AP derived from the 41 kDa precursor AP, itself the release product from the full-length 66-kDa assemblin. The 34-kDa protein probably represents the product of the inframe co translational gene within ORF 33 encoding amino acids 304-605. The genetic organisation and proteolytic maturation of VZV assemblin are, therefore, analogous to those of other herpesviruses. PMID- 9407381 TI - Characterization of hepatitis E virus (HEV) from Algeria and Chad by partial genome sequence. AB - The purpose of this study was to analyze partial nucleotide sequences and derived peptide sequences of hepatitis E virus (HEV) from two outbreaks of hepatitis E in Africa (Chad 1983-1984; Algeria 1978-1980). A portion of ORF3 and the major portion of ORF2 were amplified by Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR). The PCR products were sequenced directly or after cloning into the pCRII vector. Sequences were then compared to the corresponding regions of reported full length HEV sequences. In the ORF2 and ORF3 regions, the homology between the Algerian and the Chad isolates at the nucleic acid level was 92 and 95%, respectively. At the peptide level the homology was 98% in both regions. In these regions, both strains are more related to Asian strains at the nucleic acid level (89 to 95%) and at the amino acid level (95 to 100%) than to the Mexico strain. At the peptide level the differences are less apparent. Both African isolates have amino acid changes in common with some reference strains although the Chad isolate has three unique changes. These African strains of HEV, based on the ORF2 and ORF3 phylogenetic trees, appear to be a distinct phylogenetic group, separate from the Mexican and Asian strains. PMID- 9407382 TI - Interspousal transmission of GB virus-C/hepatitis G virus: a comparison with hepatitis C virus. AB - Although infection with GB virus-C/hepatitis G virus (GBV-C/HGV) by blood transfusion is well documented, little is known about the other routes of transmission. The prevalence of GBV-C/HGV infection in spouses of index patients and the related risk factors were studied. Hepatitis C virus (HCV) and GBV-C/HGV infections were studied in spouses of 100 patients with hepatitis C, of whom 12 were found to be also positive for GBV-C/HGV RNA. For couples both with GBV-C/ HGV viremia, nucleotide sequences of the divergent envelope region were analyzed by phylogenetic tree constructions. For HCV infection, anti-HCV was found in 14 (14%) of the 100 spouses. Five spouses (42%) of the 12 patients with dual infection of GBV-C/HGV and HCV had evidence of GBV-C/HGV infection, three had viral RNA, and two had antibodies to a recombinant HGV envelope protein E2. Nucleotide sequence comparison and phylogenetic tree analysis of the genome in the GBV-C/HGV infected couple revealed the isolates to be closely related. These results suggest that spouses of patients with GBV-C/HGV infection are at a higher risk of acquiring GBV-C/HGV as compared with HCV, and they should be educated to avoid GBV-C/HGV infection from their spouses, in case GBV-C/HGV is shown to be pathogenic. PMID- 9407383 TI - Antibody to GBV-C second envelope glycoprotein (anti-GBV-C E2): is it a marker for immunity? AB - The clinical significance of GB virus C (GBV-C E2) antibody is under investigation. The prevalence rates of GBV-C RNA and antibody to GBV-C E2 glycoprotein were determined in a population of 123 Egyptian anti-hepatitis C virus (HCV)-positive patients with chronic liver disease (CLD) who had not been treated previously with interferon. Sera were tested for GBV-C RNA by the LCx assay (Abbott Laboratories, North Chicago, IL), and for GBV-C E2 antibody by enzyme immunoassay. GBV-C RNA was present in 11.4% of patients. GBV-C E2 antibody was detected in 55.9% of GBV-C RNA-negative patients and in 2.2% of GBV-C RNA positive patients (P = 0.006). GBV-C RNA was associated significantly with a history of schistosomiasis (relative risk [RR] = 5.83, 95% confidence interval [CI] 1.99-17.14, P < 0.005) but not with parenteral risk factors. The presence of GBV-C E2 antibody was not associated with age, gender, parenteral risk factors, schistosomal infection, or HCV viremia. The HCV genotype and level of viremia were similar in GBV-C anti-E2-positive and negative patients. There was a trend toward more severe histological disease with anti-E2 seropositivity (RR = 1.45, 95% CI 0.89-2.45, P = 0.11), an association which was independent of the evidence of schistosomiasis. It is concluded that GBV-C infection is common among HCV infected Egyptian patients with CLD due to HCV infection. A significant negative correlation between the GBV-C viremia and GBV-C E2 antibody suggests that an antibody response is associated with viral clearance. This antibody response presumably occurs spontaneously, as none of the patients had received antiviral therapy. The unexpected association between GBV-C RNA and schistosomiasis suggests that nonparenteral or occult parenteral routes of GBV-C infection are likely to be important. PMID- 9407384 TI - Mutations in the nonstructural protein 5A gene and response to interferon therapy in young patients with chronic hepatitis C virus 1b infection. AB - A region associated with sensitivity to interferon (IFN) has been identified previously in the nonstructural protein 5A (NS5A) of hepatitis C virus (HCV) genotype 1b. A study was undertaken to determine whether the presence of mutations in the NS5A2209-2248 sequence could serve as a predictor of response to IFN therapy in children and adolescents with chronic HCV-1b infection. Sixteen children (M/F ratio = 11:5; mean age 11.7 years, range 5 to 19 years) with chronic HCV-1b infection who received IFN-alpha for 6 months (total dose: 8 MU/kg) were enrolled in this study. Twelve of the children (75%) had an underlying malignant disease. Pretreatment NS5A gene sequences were detected by reverse transcription-nested polymerase chain reaction (RT-nested PCR). PCR products were subjected to direct sequencing by the dideoxy chain termination method. The amino acid sequences of NS5A2209-2248 were compared with the published NS5A2209-2248 sequences of HCV-J. The NS5A2209-2248 sequences were detected in 10(63%) of the 16 children. Eight patients had the wild-type sequences, with no amino acid changes; and two patients had the intermediate type, with only one amino acid change. Four (25%) of the 16 patients responded completely to IFN therapy. Three of the four patients had the wild-type sequences, while none of the patients with the mutant type had a complete response. Serum HCV RNA levels in children with the wild type did not differ from those in patients with the mutant type. This study shows that there is no significant correlation between response to IFN and mutations in NS5A2209-2248. The amino acid sequences in NS5A2209-2248 in young patients with chronic HCV-1b infection appear to be conserved. PMID- 9407385 TI - Improved detection of respiratory syncytial virus in nasal aspirates by seminested RT-PCR. AB - A seminested RT-PCR for amplification of Respiratory syncytial virus (RSV)-RNA in nasal aspirates has been developed and used to test nasopharyngeal aspirates (NPAs) from 132 infants hospitalized with acute respiratory tract infections during winter epidemics. The results were compared with those obtained by virus isolation in tissue culture and antigen detection with an enzyme-linked immunosorbent assay (Ag-ELISA). RSV-RNA was detected by seminested RT-PCR in 57 of the 59 samples that were positive by virus isolation and/or ELISA, as well as in 25 of 73 samples negative by virus isolation and ELISA. Eighteen of these 25 samples were obtained from children older than one year of age, 17 of whom were experiencing reinfection, as indicated by the presence of preexisting serum RSV IgG antibodies. These results indicate that seminested RT-PCR is more sensitive than conventional methods for the detection of RSV in patients experiencing reinfections and suggest that this assay might also be useful for rapid diagnosis of RSV infections in older people. PMID- 9407386 TI - Correlation of patient immune responses with genetically characterized small round-structured viruses involved in outbreaks of nonbacterial acute gastroenteritis in the United States, 1990 to 1995. AB - Small round-structured viruses (SRSVs) are a genetically and antigenically diverse group of caliciviruses that are the most common cause of outbreaks of acute nonbacterial gastroenteritis. We have applied both molecular techniques to characterize SRSVs in fecal specimens and serologic assays using four different expressed SRSV antigens to examine the distribution of outbreak strains in the United States and determine if the immune responses of patients were strain specific. Strains from 23 outbreaks of SRSV gastroenteritis were characterized by reverse transcription-PCR and nucleotide sequencing of a 277-base region of the capsid gene. These strains segregated into two distinct genogroups, I and II, comprising four and six clusters of strains respectively, each representing a distinct phylogenetic lineage. Serum IgG responses in patients were measured by enzyme immunoassay using expressed capsid antigens of Norwalk virus (NV), Toronto virus (TV), Hawaii virus (HV), and Lordsdale virus (LV), representing four of the 10 clusters. While strains in genogroups I and II were antigenically distinct, within genogroups, the specificity of the immune response varied greatly. Patients infected with genogroup I strains which had as much as 38.5% aa divergence from NV demonstrated relatively homologous seroresponses to the single NV antigen. In contrast, in genogroup II, homologous seroresponses to TV and HV were only present when the infecting strains showed less than 6.5% aa divergence from these antigens. These results suggest that TV and HV represent not only separate genetic clusters in genogroup II but also separate antigenic groups, each of which is related but distinguishable. In addition, two genetically distinct SRSV strains were identified for which we have no homologous antigen. This study suggests that while current molecular diagnostics are capable of detecting the full range of SRSVs, additional expressed antigens will be required to detect an immune response to SRSV infection caused by all the antigenically diverse strains. PMID- 9407387 TI - Elevation of soluble CD23 in sera from patients with infectious mononucleosis. AB - CD23 is induced in B cells upon infection by Epstein-Barr virus (EBV) and a soluble form (soluble CD23: sCD23) is found in culture supernatants from EBV transformed B cell lines. Based on these observations, we measured serum sCD23 levels in patients with infectious mononucleosis (IM) caused by EBV infection. Sera from patients with IM at the time of diagnosis contained more sCD23 than sera from normal control subjects. Changes in serum sCD23 levels during the course of disease showed that serum sCD23 levels were elevated at the time of diagnosis and they decreased to the normal levels during the convalescent phase defined by the improvement of symptoms of IM. These results indicate that the elevated levels of sCD23 were observed at the acute phase of IM and may be useful in diagnosing IM. PMID- 9407388 TI - Gene delivery to rat enteric neurons using herpes simplex virus-based vectors. AB - Neurons of the enteric (gut) nervous system can be cultured in vitro and readily survive transplantation into the brain making close connections with host neurons. As such, they could potentially be used to deliver therapeutic gene products to the brain after transduction with appropriate genes in culture. Here the authors report the first example of gene delivery to such cultured neurons using herpes simplex virus based vectors. They show that viruses lacking the immediate early gene encoding ICP27 (which are unable to replicate lytically) can efficiently deliver a marker gene to enteric neurons without producing extensive cellular damage. In contrast, viruses lacking only the viral neurovirulence factor encoded by ICP34.5 are inefficient in gene delivery, and produce extensive cellular damage, although they cannot replicate lytically in enteric neurons. A virus lacking both ICP27 and ICP34.5, however, produces less cellular damage than one lacking only ICP27, and is as efficient in gene transfer, whereas inactivation of VMW65 reduces toxicity further. The identification of this virus as a safe and efficient gene delivery vector for enteric neurons paves the way for the eventual delivery of therapeutic genes and subsequent transplantation of engineered neurons into the CNS. PMID- 9407391 TI - Doxycycline treatment reduces ischemic brain damage in transient middle cerebral artery occlusion in the rat. AB - Agents that inhibit leukocyte adhesion including intercellular adhesion molecule 1 antibodies (anti-ICAM-1) have shown beneficial effects in experimental central nervous system (CNS) ischemia. Doxycycline inhibits leukocyte function in vitro by binding divalent cations and reduces spinal cord reperfusion injury. The authors used a clinically relevant model of focal CNS reperfusion injury to test whether treatment with doxycycline would reduce cerebral ischemic damage and improve functional outcome. Reversible middle cerebral artery occlusion was produced in adult Sprague-Dawley rats by advancing a filament into the internal carotid artery for 2 h. Animals received either i.p. doxycycline (10 mg/kg) (N = 13) or saline (N = 11) 30 min before ischemia, followed by 10 mg/kg every 8 h x 6. Both functional assessment (5 point neurologic scale) and infarct volume was evaluated at 48 h. Functional efficacy: doxycycline 0.5 +/- 0.2 (mean +/- SE) vs control 1.3 +/- 0.3 (p = 0.03). Infarct volume: doxycycline 56 +/- 18 mm3 vs control 158 +/- 44 mm3 (p = 0.03); This protective effect supports the role of doxycycline in reducing CNS reperfusion injury. PMID- 9407390 TI - High affinity neurotensin receptor mRNA distribution in rat brain and peripheral tissues. Analysis by quantitative RT-PCR. AB - Neurotensin (NT) is widely distributed in the central nervous system (CNS) and peripheral tissues, and its actions are mediated by a specific family of G protein-coupled receptors. In this study, the authors have measured the levels of gene expression of the high-affinity neurotensin receptor (NTR) with quantitative reverse-transcriptase-polymerase chain reaction (RT-PCR). In the rat brain, the highest quantities of NTR mRNA were found in the ventral mesencephalon and in the hypothalamus. Surprisingly, almost identical quantities were detected in both structures, despite results from in situ hybridization studies revealing a low expression of NTR mRNA in the hypothalamus. The RT-PCR data suggest that large scale NTR mRNA synthesis is occurring in restrictive hypothalamic nuclei. Intermediate levels of expression were detected in the prefrontal cortex and striatum, and scant levels in the cerebellum. In peripheral tissues, the highest levels of NTR mRNA were detected in the colon, followed by the liver, and then duodenum and pancreas. In this study, the sensitivity and the accuracy of the quantitative RT-PCR method provided the means to estimate the relative distribution of NTR mRNA between brain structures and peripheral tissues. Therefore, this study promotes a better understanding of the localization of NTR synthesis in relationship with the various physiological effects of NT. PMID- 9407389 TI - Melatonin alters the metabolism of the beta-amyloid precursor protein in the neuroendocrine cell line PC12. AB - The deposition of amyloid plaques in brain parenchyma is one of the major pathological hallmarks of Alzheimer's disease (AD). The amyloid in senile plaques is composed of the amyloid beta-peptide (A beta) of 39-43 amino acid residues derived from a larger beta-amyloid precursor protein (beta APP). Soluble derivatives of beta APP (sAPP) lacking the cytoplasmic tail, transmembrane domain, and a small portion of the extracellular domain are generated proteolytically by "secretases." Using cell cultures, the authors analyzed the level of sAPP in neuroblastoma and pheochromocytoma (PC12) cells by immunoblotting samples from conditioned media and cell lysates. Normal levels of secretion of sAPP into conditioned media were severely inhibited by treating cells with melatonin (3-4 mM). The inhibitory effect of melatonin on the secretion of sAPP can be reversed. When the cells that were pretreated with melatonin for 10 h were washed, the normal level of secretion of sAPP was restored. Northern blot analyses indicated that the treatment of PC12 cells with melatonin resulted in a significant decrease in the level of mRNA encoding beta APP, beta-actin, and glyceraldehyde-3-phosphate dehydrogenase, and that the treatment of a human neuroblastoma cell line with melatonin resulted in no change in levels of these messages. The secretion of sAPP into the conditioned medium was substantially reduced in the differentiated cells similar to reductions observed in melatonin-treated undifferentiated PC12 cells. Melatonin was found to potentiate the nerve growth factor-mediated differentiation in PC12 cells at 24 h. Taken together, these data suggest that melatonin regulates the metabolism of beta APP and other housekeeping genes in a cell-type specific manner, and that melatonin accelerates the early process of neuronal differentiation. PMID- 9407392 TI - Canavan disease. Analysis of the nature of the metabolic lesions responsible for development of the observed clinical symptoms. AB - Canavan disease (CD), a rare recessive autosomal genetic disorder, is characterized by early onset and a progressive spongy degeneration of the brain involving loss of the axon's myelin sheath. After a relatively normal birth, homozygous individuals generally develop clinical symptoms within months, and usually die within several years of the onset of the disease. A biochemical defect associated with this disease results in reduced activity of the enzyme N acetyl-L-aspartate amidohydrolase (aspartoacylase) and affected individuals have less ability to hydrolyze N-acetyl-L-asparate (NAA) in brain and other tissues. As a result of aspartoacylase deficiency, NAA builds up in extracellular fluids (ECF) and is excreted in urine. From an analysis of the NAA biochemical cycle in various tissues of many vertebrate species, evidence is presented that there may be two distinct NAA circulation patterns related to aspartoacylase activity. These include near-field circulations in the brain and the eye, and a far-field systemic circulation involving the liver and kidney, the purpose of which in each case is apparently to regenerate aspartate (Asp) in order for it to be recycled into NAA as part of the still unknown function of the NAA cycle. Based on the authors' analysis, they have also identified several metabolic outcomes of the genetic biochemical aspartoacylase lesion. First, there is a daily induced Asp deficit in the central nervous system (CNS) that is at least six times the static level of available free Asp. Second, there is up to a 50-fold drop in the intercompartmental NAA gradient, and third, the ability of the brain to perform its normal intercompartmental cycling of NAA to Asp is terminated, and as a result, the only remaining long-term source of Asp for NAA synthesis is via nutritional supplementation of Asp or its metabolic precursors. Finally, the authors identify a potential maternal-fetal interaction that may be responsible for observed normal fetal development in utero, and that provides a rationale for, and suggests how, CD might respond to far-field nutritional, transplantation, or genetic engineering techniques to alter the course of the disease. PMID- 9407394 TI - Tetracycline fibers plus scaling and root planing versus scaling and root planing alone: similar results after 5 years. AB - This paper presents 5-year data pertaining to a subgroup of patients from a previous investigation who were treated with scaling and root planing plus tetracycline fibers. The parent study demonstrated that 6 months after therapy, scaling and root planing plus tetracycline fiber therapy was significantly better at reducing probing depth and gaining clinical attachment than scaling and root planing alone. However, the long-term data presented here show a regression from the original gains in clinical attachment levels in the fiber group. Ultimately, the use of fibers provided no significant advantage with regards to probing depth reduction or clinical attachment gain. Within the power of this study, which would have required 1.78 mm of change in clinical attachment to show a difference, there was no significant difference between the treatments at 5 years. This study underscores the need for additional long-term evaluation of this mode of therapy. PMID- 9407393 TI - Pituitary adenylate cyclase activating polypeptide (PACAP) regulates expression of catecholamine biosynthetic enzyme genes in bovine adrenal chromaffin cells. AB - Pituitary adenylate cyclase activating polypeptide (PACAP) elevates levels of the mRNAs encoding the catecholamine synthesizing enzymes tyrosine hydroxylase (TH), dopamine beta-hydroxylase (DBH), and phenylethanolamine N-methyltransferase (PNMT) in primary cultures of bovine adrenal chromaffin cells. PACAP potently (in nanomolar concentrations) increases the amount of mRNA for each of the three catecholamine biosynthetic enzymes. At 10 nM PACAP, TH and DBH mRNA levels increase approx 10-fold; 1 nM PACAP produces an approx 2.5-fold elevation of PNMT mRNA. In contrast to depolarizing or cholinergic stimuli, PACAP does not enhance expression of 5' upstream regions of the PNMT gene transiently transfected into chromaffin cells. Nor does PACAP stimulate the rate of PNMT gene transcription, thereby indicating that the effects of this neuropeptide do not involve enhanced transcription of this gene. However, after 16 h in the presence of transcriptional inhibitors, more PNMT mRNA is present in cultures treated with PACAP relative to control cultures, whereas amounts of TH and DBH mRNAs are not changed. PACAP likely elevates PNMT mRNA levels posttranscriptionally, possibly by stabilizing this message against degradation. Thus, although PACAP is an effective regulator for expression of all three catecholamine enzyme genes, its mechanism of action on PNMT mRNA appears to be distinctive from its effects on TH and DBH gene transcription. PMID- 9407395 TI - A 10-year study of the progression of destructive periodontal disease in adult and elderly Chinese. AB - This study describes the progression of destructive periodontal disease among Chinese aged 20 to 80 with limited access to dental health facilities and minimal traditions for oral hygiene procedures. These individuals were followed for 10 years to determine whether the rates for progression of periodontal disease were markedly different than for populations with more access to oral health care. At baseline, participants had been examined for tooth mobility, plaque, calculus, gingival conditions, attachment levels, and probing depths on 4 sites of each tooth present. These probing depth and attachment level recordings were repeated at follow-up, although third molars were excluded from examination. A total of 398 persons remained dentate at follow-up. The analysis demonstrated that virtually all subjects experienced > or = 2 mm attachment loss over the 10-year period, and frequently in a large proportion of the sites present. Attachment loss > or = 3 mm was also widespread, but the distribution of persons according to the extent of > or = 3 mm attachment loss was positively skewed in all age groups. Positive skewness was even more pronounced when attachment loss of > or = 4 mm was considered. Some types of teeth, such as mandibular incisors and maxillary molars, had higher progression rates than did, for example, maxillary incisors. The mean individual attachment loss rates did not differ significantly between age groups, and were remarkably similar to those reported for populations whose access to and tradition for oral health care is widespread. PMID- 9407396 TI - Comparison of porous bone mineral and biologically active glass in critical-sized defects. AB - Several materials have been proposed as therapies to augment alveolar bone and to promote periodontal regeneration. However, there are an insufficient number of studies that effectively evaluated these therapies. Consequently, the purpose of this study was to compare bone regeneration promoted by porous bone mineral and biologically active glass. Unilateral critical-sized defects (CSDs) were prepared in the radii of 24 rabbits, divided evenly between 2 time periods (4 and 8 weeks) and between 2 treatment groups (porous bone mineral and biologically active glass). Evaluations consisted of clinical examinations, standardized radiography at baseline and every 2 weeks thereafter, as well as histology and histomorphometry. Data were analyzed by an unpaired Student t-test with significance established at P < or = 0.05. We determined that CSDs treated with porous bone mineral were significantly more radiopaque than biologically active glass-treated sites at both 4 and 8 weeks. Moreover, the amount of new bone was significantly greater at both 4 and 8 weeks in the porous bone mineral groups than in the biologically active glass groups. We concluded that in the rabbit radius CSD wound model, porous bone mineral appears to be more effective than biologically active glass in regenerating bone. PMID- 9407397 TI - Development and characterization of a transformed human periodontal ligament cell line. AB - Periodontal ligament (PDL) cells are thought to be important for establishing and maintaining a stable interface between bone and teeth. In addition, PDL cells are thought to play critical roles in both the pathogenesis of periodontal disease and the regeneration of periodontal ligament tissues. The purpose of this study was to develop a continuous or stable human PDL cell line as an in vitro model for the investigation of cellular mechanisms involved in periodontal regeneration and destruction. Human PDL cells, derived from a primary cell culture, were transfected with simian virus 40 (SV40) T antigen-containing virus with a neomycin resistance gene. The transformed cells expressed the SV40 T antigen mRNA as assayed by reverse transcription polymerase chain reaction (RT-PCR). This cell line was also characterized for morphological changes and growth characteristics compared to primary PDL cell cultures. The transformed cells were shown to form a multilayer pattern and distinct colonies on tissue culture surfaces. However, no colony formation was found in soft agar. The transformed PDL cell line was found to have a greater rate of proliferation in 10% fetal bovine serum than primary culture, and continued to proliferate in low serum concentrations capable of producing quiescence in primary cells. Interleukin-1 beta (IL-1 beta) was shown to produce a 7-fold elevation in collagenase (MMP-1) mRNA levels, consistent with primary PDL cells. In addition, IL-1 beta was shown to produce a decrease in alkaline phosphatase activity in a concentration-dependent manner. The transformed cell line has been maintained for over 30 generations of cell culture. In conclusion, a stable human PDL cell line has been established to serve as a model for future in vitro investigations into periodontal pathogenic mechanisms and to evaluate therapies directed at the regeneration of periodontal ligament. PMID- 9407398 TI - An investigation of preferential fibroblast wound repopulation using a novel in vitro wound model. AB - To overcome the difficulties of studying wounding and wound repopulation in monolayer systems, a 3-dimensional model of wound repopulation has been developed which allows the in vitro investigation of fibroblast migration in response to experimental wounding. This model was utilized to determine whether fibroblasts derived from sites which demonstrate preferential healing (child and oral mucosal fibroblasts) possessed an increased ability to repopulate experimental wounds when compared to adult dermal fibroblasts. Fibroblasts were established from specimens derived from healthy donors undergoing minor elective surgery. Standard wounds were created in fibroblast populated collagen lattices (FPCLs) which were then overlaid upon an extracellular wound matrix. Fibroblast repopulation of the wounds was studied over 12 days using light- and scanning electron microscopy and quantified using computerized image analysis. Wound repopulation by fibroblasts derived from child donors (n = 3) was significantly (P < 0.001) more rapid than their adult tissue-matched counterparts (n = 3). Wound repopulation by oral mucosal fibroblasts (n = 3) was significantly greater than that exhibited by age matched dermal fibroblasts (n = 3; P < 0.05). These differences were not reflected in differences in DNA synthesis (P > 0.5) or cell number (P > 0.5) within similar attached FPCL systems. These findings further support the concept of a gradual transition from the fetal to adult phenotype in wound healing. The potential applications of the model are discussed. PMID- 9407399 TI - Effect of inflammation on the proliferation of human gingival epithelial cells in vitro. AB - The adaptive or pathologic responses of epithelial cells to inflammation are poorly characterized. The purpose of this study was to determine if epithelial cells cultured from clinically healthy and inflamed human gingival tissues express differences in proliferation rate and viability. Briefly, the inflammation status of individual donor sites from 101 patients was visually assessed at the time of periodontal surgery and categorized as either non-to slightly inflamed, moderately inflamed, or severely inflamed. Discarded gingival tissues were then processed to obtain primary cell cultures, for which proliferation rates were determined by calculating the ratio of mean population doublings to the number of days required for cultures to become confluent. In general, the cells in the minimally inflamed group exhibited characteristics different than cells in the moderately and severely inflamed groups. Specifically, the cells obtained from clinical sites which exhibited no-to-slight inflammation had a significantly higher mean proliferation rate than cells in either the moderate inflammation group or the severe inflammation group. Based on trypan blue exclusion, the cells obtained from clinical sites which exhibited no to-slight inflammation also were more viable than cells obtained from sites with moderate inflammation or severe inflammation. Microscopic evaluation showed morphological changes associated with increased inflammation. Cell cycle analysis by fluorescent-activated cell sorting (FACS) revealed a directly proportional relationship between the degree of inflammation and apoptosis, and a strong inversely proportional trend between the degree of inflammation and the numbers of cells undergoing mitosis. Taken together, these data suggest that epithelial cell proliferation and viability are inversely associated with the degree of gingival inflammation, once a putative "adaptive threshold" is exceeded. Elucidation of the underlying mechanisms will likely lead to improvements in clinical diagnosis and treatment. PMID- 9407400 TI - A quantitative assessment of osteoinductivity of human demineralized bone matrix. AB - Demineralized bone matrix (DBM) is widely used in the repair of pathologies associated with skeletal defects and periodontal diseases. The present study was directed at establishing in vivo and in vitro models for a quantitative assessment of the osteoinductivity of DBM before clinical use. Athymic mice were used in an in vivo assay to overcome the species limitations (for human DBM) found in xenogeneic animal models. Calcium contents of explants, as an indicator of new bone formation, were assayed and expressed as a change in the weight percent calcium in the explant as compared to the weight percent of calcium in the implanted material. A total of 82 mice (2 implants per mouse) were used in this study. Significant amounts of new bone were induced in this animal model in response to implantation of DBM. Muscular implantation was found to be more osteoinductive (increases of 10.0 +/- 0.4 calcium weight percent of explant) than subcutaneous implantation (increases of 1.62 +/- 0.27 calcium weight percent of explant) and new bone formation in muscular implantation sites of athymic mice mimics endochondral bone formation. Between weeks 1 to 4, the weight of explanted materials did not significantly differ from the weight of the implanted material; however, by week 5 the explant weight began to increase. Calcium deposition over the 5 weeks of implantation increased in a nearly linear fashion. Consequently week 4 was chosen as the optimum time for explantation in the in vivo assay in that sufficient calcium levels had been achieved without a significant increase in explant dry weight. Aliquots of 10, 20, 30, and 40 mg per implantation site were used in dose response studies in the in vivo bioassay. Dose response curves with DBM exhibited maximal activity at the 20 mg DBM implant dose in the in vivo bioassay. An in vitro bioassay was also developed where human periosteal (HPO) cells were chosen because osteoprogenitor cells found in bone repair typically come from periosteal tissue. Alkaline phosphatase (ALP) activity in confluent cell cultures of HPO cells exposed to DBM, as an indicator of osteoblast induction, reached its highest level on day 5 of DBM treatment. Aliquots of 2, 5, 10, 20, 30, and 40 mg DBM per flask were chosen in dose response studies using the in vitro bioassay. These dose response studies with DBM revealed that quantities approximating 5 to 10 mg DBM in the in vitro model provided for maximal levels of ALP in cell extracts. A linear correlation (R2 = 0.7397) was demonstrated between the in vivo calcium remineralization assay and the in vitro ALP assay of osteoinductivity of DBM, suggesting that the in vitro assay can be used to quantitatively assess the osteoinductive potential of DBM where production and distribution of clinically usable DBM dictates rapid analysis. PMID- 9407401 TI - Effect(s) of the demineralization process on the osteoinductivity of demineralized bone matrix. AB - The relationships between residual calcium levels and particle size of ground demineralized bone matrix and its osteoinductive potential were investigated using in vitro and in vivo assays. The effects of variable residual calcium levels, variable particle sizes, and donor age and gender were studied using a tissue culture-based bioassay (in vitro) as well as an athymic mouse (in vivo) bioassay. The osteoinductive potential of the bone-derived biomaterial was assessed by measuring the degree of new bone formation (change in percent calcium content after 4 weeks of implantation) in the in vivo assay and levels of alkaline phosphatase activity associated with cultures of human periosteal cells (HPO cells) in the in vitro assay, respectively. Slightly demineralized bone matrix and overly demineralized bone matrix possessed a degree of osteoinductive potential whereas bone demineralized to levels of approximately 2% residual calcium provided for maximum osteoinductive potential in both assay systems. The osteoinductive potential of ground demineralized bone varied relative to the particle size such that DBM particles ranging from 500 to 710 microns provided for the highest level of calcium deposition (increase of 8.1 weight percent calcium) after 4 weeks of implantation in muscle pouches of an athymic mouse, whereas explanted particles less than 250 microns showed the lowest level of calcium deposition (increase of only 2.8 weight percent calcium). In the donor age and gender study, DBM from different donors were divided into 5 age groups for both female and male donor derived bone: less than 20, 21 to 30, 31 to 40, 41 to 50, and 51 to 60 year old age groups. This study indicated that DBM from female donors in the 31 to 40 years old age group and male donors in the 41 to 50 year age group possess the highest osteoinductive potential, whereas DBM derived from donor bone from both female and male donors in the 51 to 60 year age group presented the lowest osteoinductive potential. DBM derived from male and female donors did not in general show significant differences in osteoinductive potential. PMID- 9407402 TI - Comparative in vitro effectiveness of closed root debridement with fine instruments on specific areas of mandibular first molar furcations. I. Root trunk and furcation entrance. AB - The purpose of this study was to compare curets with a small blade to slim ultrasonic inserts on their efficacy in removing artificial deposits from the root trunk and furcation entrance areas of mandibular molars using an in vitro model simulating a clinically closed root debridement approach. The study was conducted on 100 artificial mandibular first molars (50 right side and 50 left side) with anatomical roots. Root trunks, furcation entrances, and furcation areas of each molar were colored by a coat of black model paint. The teeth were fixed in a custom acrylic model and maintained in a firm position by modified acrylic occlusal splints. The root areas were covered with a heavy rubber dam imitating gingival tissue. The model was attached to a mannequin and mounted on a dental chair. Fifty molars (25 right, 25 left) were instrumented with the experimental curets and an equivalent number of molars with the ultrasonic inserts. The instrumentation was carried out by one experienced operator, spending 4 minutes on each molar. The instrumented areas were individually analyzed to determine the percentage of deposits remaining, using a computerized imaging routine system. One-way analysis of variance was conducted to test for differences between both types of instruments. Results revealed that the curets were significantly more efficient (P < 0.01) than the ultrasonic inserts in removing paint from both root trunks and furcation entrances. These findings should be corroborated in a clinical study to determine the potential value of the instruments tested during initial therapy or supportive care of involved mandibular furcations. PMID- 9407403 TI - Comparative in vitro effectiveness of closed root debridement with fine instruments on specific areas of mandibular first molar furcations. II. Furcation area. AB - The purpose of this study was to demonstrate the extent of deposits removed from within the furcation area of mandibular first molars following the use of curets with a modified blade and slim ultrasonic inserts in an in vitro model simulating a closed root debridement approach to furcation treatment. The furcation areas of 100 artificial mandibular first molars were uniformly coated with black model paint. The molars were fixed into a custom acrylic model, maintained in a firm position with modified occlusal splints, and the roots covered with a heavy rubber dam. The model was set in a mannequin and mounted on a dental chair recreating a clinical situation. Fifty molars (25 right, 25 left) were instrumented with the experimental curets and an equivalent number of molars with the ultrasonic inserts. An experienced dental hygienist completed all the instrumentation, spending 4 minutes on each molar. The molars were sectioned buccolingually from the crown apically to separate the roots, and areas in the internal surface of mesial and distal roots were analyzed to determine the percentage of deposits remaining using a computerized imaging routine system. A 2 factor analysis of variance was conducted to test for differences between both types of instruments. The curets produced furcation root surfaces with significantly less percentage of residual deposits than the ultrasonic inserts (P < 0.01). This study indicates the potential value of small bladed curets in debriding involved furcations during initial therapy and supportive periodontal therapy. The current findings should be corroborated in a clinical study. PMID- 9407404 TI - Clinical and microbiological effects of minocycline-loaded microcapsules in adult periodontitis. AB - Clinical and microbiological effects of subgingival delivery of 10% minocycline loaded (MC), bioabsorbable microcapsules were examined in 15 adult periodontitis patients. Patients received oral hygiene instruction 2 weeks prior to the study. At baseline (day 0) all teeth received supragingival scaling (SC); 2 quadrants received no further treatment and 1 quadrant received subgingival scaling and root planning (SRP). In the fourth quadrant, the tooth with the deepest probing sites (at least 1 site > or = 5 mm) was treated with minocycline microcapsules. The sites were evaluated at baseline and weeks 1, 2, 4, and 6. Clinical indices included bleeding on probing (BOP), probing depths (PD), and attachment loss (AL). Microbiological evaluations included percent morphotypes by phase-contrast microscopy; cultivable anaerobic, aerobic, and black-pigmented Bacteroides (BPB); and percent Porphyromonas gingivalis, Prevotella intermedia, Eikenella corrodens, and Actinomyces viscosus by indirect immunofluorescence. In the SC + MC group, BOP, PD, and AL were significantly reduced from baseline for weeks 1 to 6. BOP in the SC + MC group was significantly reduced compared to the SRP group from weeks 2 to 6. In the SC + MC group the percent of spirochetes and motile rods decreased and the percent of cocci increased after 1 week. The increased cocci and decreased motile rods were statistically greater at weeks 4 and 6 in the SC + MC group compared to the SRP group. This study demonstrates that local subgingival delivery of 10% minocycline-loaded microcapsules as an adjunct to scaling results in reduction in the percent sites bleeding on probing greater than scaling and root planning alone and induces a microbial response more favorable for periodontal health than scaling and root planing. PMID- 9407405 TI - Placement of implants into fresh extraction sites: 4 to 7 years retrospective evaluation of 95 immediate implants. AB - A 7-year follow-up of implants placed immediately after tooth extraction into fresh extraction sites is reported. Small autogenous bone chips (from bone adjacent to implant sites) were grafted into the defect between the implant and the socket walls when needed. Closure of the wound was obtained by coronal repositioning of the flap, and no membranes were used. Care was taken to minimize hematoma formation under the flap during healing by part-time use of removable prosthesis with thick soft linings after implant surgery. At second stage surgery, mucoperiosteal flaps were apically repositioned for maximum attached gingival width and to reconstruct the vestibule. Minor complications such as exposure occurred in 16% of cases. Implant mean 5-year cumulative survival rate was 95%. There was no implant loss after loading. The results indicated that implants placed into fresh extraction sites grafted with autogenous bone chips will heal predictably. PMID- 9407406 TI - Crestal bone changes around titanium implants. A radiographic evaluation of unloaded nonsubmerged and submerged implants in the canine mandible. AB - Current implant placement utilizes both nonsubmerged and submerged techniques. However, the implications of the location of a rough/smooth implant interface as well as the location of a microgap between implant and abutment on crestal bone changes are not well understood. The purpose of this study was to radiographically evaluate crestal bone changes around unloaded nonsubmerged and submerged titanium implants in a side-by-side comparison. Fifty-nine (59) implants were placed at different levels to the alveolar crest in 5 foxhounds. Standardized radiographs were taken at baseline and at monthly intervals until sacrifice at 6 months. Radiographic assessment was carried out by measuring the distance between the top of the implant/abutment and the most coronal bone-to implant contact (DIB), and by evaluation of bone density changes using computer assisted densitometric image analysis (CADIA). DIB measurements revealed that in 1-part, nonsubmerged implants, the most coronal bone-to-implant contact followed at all time points the rough/smooth implant interface. In all 2-part implants, nonsubmerged and submerged, the most coronal bone-to-implant contact was consistently located approximately 2 mm below the microgap. In addition, CADIA values for all 2-part implants were decreased in the most coronal area-of interest (AOI). All bone changes were statistically significant and detectable 1 month after implant placement in nonsubmerged implants or 1 month after abutment connection in submerged implants. Neither implant position nor individual dog effects were statistically significant. These results demonstrate that the rough/smooth implant interface as well as the location of the microgap have a significant effect on marginal bone formation as evaluated by standardized longitudinal radiography. Bone remodeling occurs rapidly during the early healing phase after implant placement for non-submerged implants and after abutment connection for submerged implants. PMID- 9407407 TI - Experimental peri-implantitis in consecutively placed, loaded root-form and plate form implants in adult Macaca mulatta monkeys. AB - This study is part of an on-going project describing the character of round- and plate-form implants placed in a primate model. In this paper 20 loaded plate-form and 23 root-form implants were connected to prostheses and experimental peri implantitis was induced by ceasing scaling procedures and placing braided silk ligature around the implants. Twenty-four prostheses utilizing natural teeth were studied for comparison. Clinical measurements were carried out monthly for 6 months and radiographic measurements at 3 and 6 months post-ligature placement and cessation of scaling. Both root-form implants and plate-form implants showed a significant loss of crestal bone height at 3 and 6 months after ligature placement (P < .001 after 6 months). The difference in bone loss between plate- and root-form implants, however, was not significant. PMID- 9407408 TI - Role of psychiatric disorders in self-inflicted periodontal injury: a case report. AB - Self-inflicted or factitious injuries (FI) are not uncommon in psychiatric patients. In general, this kind of behavior is linked to secondary gain. However, the role of underlying psychiatric illness should not be overlooked. Usually, the diagnosis of factitious injury can be confirmed by a careful medical-dental history, clinical appearance of the lesion, laboratory investigations, and response to established treatment protocol. A case is presented in which repetitive injurious behavior resulted in rapid periodontal attachment loss. The lesion responded well to conservative periodontal treatment. The role of underlying psychiatric morbidity leading to repetitive self-injurious behavior is discussed. PMID- 9407410 TI - Cytokines, growth factors and renal injury: where do we go now? AB - The renal response to injury is mediated by a wide variety of cytokines and growth factors. This review provides a simplified schema in which the mediators are grouped into four categories: proinflammatory cytokines, vasoactive mediators, growth factor/matrix modulating molecules, and protease/matrix proteins that modulate cell behavior. Included are examples of how each group of mediators affect renal injury. Also discussed are the problems of modulating the cytokine response, including the problems of compensation or redundancy, modulating cytokines that have multiple actions, the consequences of partial versus complete blockade of a cytokine, and modifying renal disease when the renal injury in heterogenous. Despite these concerns, the recent progress in cytokines in disease pathogenesis is very likely to lead to new treatments of kidney disease. PMID- 9407409 TI - Periodontal management of gingival overgrowth in the heart transplant patient: a case report. AB - Heart transplant patients take several medications that could affect their periodontal health. Gingival overgrowth associated with cyclosporin (immunosuppressant agent) and nifedipine (calcium channel blocker) is well documented. Candidal infections often develop because of immune suppression. This report describes the clinical and histopathological changes in the gingival tissues of a heart transplant patient and their management. The gingival tissues exhibited pronounced enlargement. The gingivae were lobulated, and the surface of the lobulations was pebbly and granular. Biopsies showed lobules of fibrous connective tissue covered by stratified squamous epithelium. The outer surfaces were dotted with numerous smaller papillations. Candidal hyphae were present in the superficial layers of the epithelium. The extensive papillary lesions appear to be related to candidiasis and constitute a condition which is best designated as papillary stomatitis. Hyperplastic gingival tissues were excised, and the patient was placed on periodic maintenance. One-year postoperative follow-up showed minor gingival growth. PMID- 9407411 TI - Role of angiotensin: insight from gene targeting studies. AB - We and others have recently produced, by gene targeting, several strains of mice deficient of a specific gene within the renin-angiotensin system. Detailed examination of these mutant mice not only confirmed the well-known blood pressure raising action of angiotensin II, but also revealed several important biological functions of angiotensin II that have not been well recognized, most notably the role in the ontogenesis of the kidney. Since gene targeting causes complete inactivation of the targeted gene, this approach also allows us to address the question, "is angiotensin really essential for the induction of aldosterone during ECF volume depletion?" In our studies, we obtained unequivocal evidence indicating that angiotensin is not essential, but instead, potassium becomes an effective regulator for aldosterone during extracellular fluid (ECF) volume depletion through mechanisms that are indistinguishable from those of angiotensin, that is, selective stimulation of aldosterone synthetase and proliferation of zona glomerulosa cells within the adrenal gland. In this context, angiotensin can be regarded as a modifier of K-dependent aldosterone regulation. PMID- 9407412 TI - Proteinuria is a risk factor for mortality over 10 years of follow-up. MRFIT Research Group. Multiple Risk Factor Intervention Trial. AB - Proteinuria has been shown to be strongly associated with the prevalence and incidence of cardiovascular disease. It has been difficult to determine if the link is causal and independent. The mortality follow-up for the Multiple Risk Factor Intervention Trial (MRFIT) randomized cohort provides an opportunity to examine these relationships. Between 1973 and 1975, 361,662 men, ages 35 to 57, were screened for blood pressure, serum cholesterol, and cigarette smoking. Patients receiving medication for diabetes were excluded. Men in the upper 10 to 15% of coronary heart disease (CHD) risk (12,866) were randomized into the MRFIT trial. Standard casual urine dipstick determinations (Labstix) for protein were done at baseline and annually for six years. Post-trial cause-specific mortality was ascertained using the National Death Index. During the trial, 2326 (18.1%) of participants had + or higher proteinuria, and 593 (4.6%) had +2 or higher proteinuria. The presence of proteinuria during the six years of follow-up was consistently associated with higher all cause, cardiovascular disease (CVD) and CHD mortality, even after adjusting for other risk factors. The higher and more persistent the proteinuria, the greater the risk. In this data set, proteinuria is a strong and independent risk factor for CVD mortality. PMID- 9407413 TI - High normo- or low microalbuminuria: basis for intervention in insulin-dependent diabetes mellitus. AB - Recent studies have further elucidated the association between blood pressure and albumin excretion in insulin-dependent diabetes mellitus (IDDM) patients with (i) normal urinary albumin excretion (UAE), and (ii) moderate microalbuminuria (20 to 70 micrograms/min). In a study comprising 117 normoalbuminuric (UAE < 20 micrograms/min) patients we performed 24-hour ambulatory blood pressure monitoring (AMBP), and short-term power spectral analysis of RR interval oscillations. In comparison with the group with a UAE below the median, patients with UAE above the median were characterized by: significantly higher 24-hour systolic and diastolic AMBP, significantly reduced short-term RR interval variability, and significantly higher HbA1c. In a double blind study, normotensive IDDM patients with moderate microalbuminuria (20 to 70 micrograms/min) were randomized to either lisinopril (20 mg once a day, N = 12) or placebo (N = 10) for two years. In the lisinopril group there were significant reductions in 24-hour systolic and diastolic AMBP compared to the placebo group. Lisinopril did not attenuate the diurnal blood pressure variation. UAE tended to be reduced in the lisinopril group. A significantly positive association between changes in AMBP and changes in UAE was present in the placebo group in contrast to the lisinopril group. Changes in UAE were strongly and positively associated with changes in filtration fraction in the lisinopril treated group. In conclusion, interactions between albumin excretion, blood pressure, autonomic function, and glycemic status are already detectable within the normoalbuminuric range in IDDM patients. Angiotensin converting enzyme inhibitor (ACEi) treatment in a small group of normotensive IDDM patients with moderate microalbuminuria reduces blood pressure without attenuating diurnal blood pressure variation, tends to reduce albumin excretion, and abolishes the association between changes in UAE and changes in blood pressure observed in the placebo group. ACEi intervention in selected normoalbuminuric high risk patients (high-normal UAE, high-normal blood pressure, and poor glycemic control) would be of interest. PMID- 9407414 TI - Comparison of renin-angiotensin to calcium channel blockade in renal disease. AB - Whether any class of antihypertensive drugs has specific renoprotective effects above and beyond lowering of blood pressure is still debatable. The renin angiotensin system (RAS) is both localized and has many actions within the kidney, on intrarenal hemodynamics, on the mesangial cell, as well as stimulating growth factors and cytokines. Angiotensin converting enzyme (ACE) inhibitors have been shown to ameliorate the progression of renal failure. How much of this beneficial effect is due to their hemodynamic effects, how much to non hemodynamic effects and how much to their effects on bradykinin and other putative ACE substrates is still unclear. Experimentally it can be shown that inhibiting ACE but preventing the fall in systemic blood pressure by salt loading abolishes renoprotection. Bradykinin has been implicated in both the beneficial and the adverse effects of ACE inhibitors. Because of this and because ACE inhibitors may not provide complete blockade of the RAS, angiotensin receptor (AT1R) antagonists have been developed. Experimentally AT1R antagonists have been shown to reproduce most of the beneficial effects of ACE inhibitors. The experience in humans is more limited but they have been demonstrated to be efficacious in hypertension, to reduce proteinuria, and produce a favorable hemodynamic effect in congestive cardiac failure with a low incidence of adverse effects and without cough. Calcium channel blockers (CCB) also have additional properties that may provide renoprotection beyond lowering blood pressure. However, as the different types of CCB block different calcium channels their effects may differ substantially. The inconsistency of the data in the renoprotective effect of CCB may reflect these differences. Quantitatively probably the most important factor in preventing the progress of renal failure by antihypertensive drugs is strict control of blood pressure. Lowering blood pressure by drugs is most likely effective by both reducing physical and sheer stress damage, as well as turning off the signal for the activation and production of vasoactive peptides and cytokines. PMID- 9407415 TI - Angiotensin converting enzyme insertion/deletion polymorphism and short-term renal response to ACE inhibition: role of sodium status. AB - Angiotensin converting enzyme (ACEi) inhibition retards renal function loss, but the therapeutic benefit varies between individuals. Renoprotection is poor in patients with the ACE DD genotype. ACE genotype is reported to affect short-term antiproteinuric response to ACEi, a predictor of long-term renoprotection, in some studies but not in others. Short-term responses to ACEi are enhanced by stimulating the renin-angiotensin system, that is, sodium restriction. We hypothesized that the ACE genotype influences sodium dependency of the response to ACEi. Therefore, we performed a cross sectional analysis of short-term responses to ACEi (enalapril or lisinopril) in 88 patients with stable non diabetic proteinuria (> 1.0 g/day) and variable sodium intake. ACE genotype distribution was: DD, N = 25; ID, N = 40; II, N = 23. Baseline proteinuria (5.9 +/- 0.7; 5.8 +/- 0.07; 4.8 +/- 0.8 g/day, respectively) and mean arterial pressure (108 +/- 3; 106 +/- 2; 107 +/- 2 mm Hg, respectively) were similar for the three genotypes. ACEi similarly reduced proteinuria (-49 +/- 5; -55 +/- 4, 48 +/- 6%, respectively) and blood pressure (-12 +/- 3; -14 +/- 1 and -12 +/- 2%, respectively) in the three groups. Interestingly, the responses to ACEi of proteinuria (r = 0.42, P < 0.05) and blood pressure (r = 0.41, P < 0.05) correlated with urinary sodium excretion in DD genotype but not in the ID (r = 0.05 and 0.17, resp) or II genotype (r = 0.09 and 0.08, respectively). Thus, in the DD group, individuals with a high sodium excretion had a less effective response to ACEi. We conclude that differences in sodium status could account for disparities between studies on the relationship between ACE genotype and response to ACEi, and that sodium restriction might be a strategy to circumvent treatment resistance in the DD genotype. PMID- 9407416 TI - Genetic determinants of diabetic renal disease and their impact on therapeutic interventions. AB - Approximately 30% of patients with type 1 and type 2 diabetes develop diabetic nephropathy. Apart from metabolic control, genetic predisposition plays an important role in its genesis. Analysis of intermediate phenotypic markers showed that the activity of Na/Li- and Na+/H(+)-countertransport is increased in patients with diabetic nephropathy. The renin-angiotensin system is of crucial importance as a system for therapeutic intervention and as genetic marker for susceptibility to renal disease. Consequently, the analysis of molecular genetic markers has focused on a polymorphism in the gene for the angiotensin II converting enzyme (ACE). However, the analysis of the I/D-polymorphism with respect to development of diabetic nephropathy in type 1 and type 2 diabetes has yielded conflicting results, at least in type 1 diabetes. These discrepant results may be due to differences in definition, sample size and ethnic background of the patients. In IgA glomerulonephritis it has been shown that the DD genotype (which is correlated with higher serum and tissue ACE activity compared to II genotype) is associated with a more rapid deterioration of renal function. The same adverse effect of the DD genotype could also be demonstrated in patients with diabetic nephropathy. Two studies examined the response to treatment according to the different genotypes, with contradictory results. A Japanese study showed a more pronounced reduction in proteinuria under ACE inhibitor treatment in patients with DD genotype, whereas a Danish study showed that patients with the DD genotype exhibited a steeper decline in renal function despite ACE inhibitor treatment. The data available for other candidate genes are fragmentary and negative throughout. PMID- 9407417 TI - Angiotensin converting enzyme inhibitors in diabetic patients with microalbuminuria or normoalbuminuria. AB - In adult diabetic patients microalbuminuria is a marker of early vascular damage in the micro- and macrocirculation. Microalbuminuria is a powerful predictor of renal and cardiovascular disease outcome and is associated with other, potentially modifiable, risk factors of vascular damage. Studies of secondary prevention have shown that blood pressure lowering drugs effectively reduce albumin excretion rate. Angiotensin converting enzyme (ACE) inhibitors seem particularly effective in reducing the risk of progression to clinical albuminuria in both insulin dependent and non-insulin dependent diabetic patients and this beneficial effect appears to be long-lasting. Whether this postpones the onset of end-stage renal failure and/or reduces early mortality in these patients remains to be established. Recent studies of primary prevention in insulin dependent diabetic patients predominantly with normoalbuminuria demonstrate that ACE inhibition reduces significantly the rate of progression of albumin excretion rate and, of great interest, seems to affect beneficially the progression of retinopathy. These results compare favorably with the beneficial effect of intensified insulin therapy and strict blood glucose control in this same group of patients. Thus, ACE inhibitors are a powerful tool to prevent progression of microalbuminuria in diabetes and may prove useful as an adjunct therapy to intensified insulin therapy in the prevention of development of microalbuminuria and of retinopathy progression in insulin dependent diabetes. PMID- 9407418 TI - Glycemic control and the initiation and progression of the complications of diabetes mellitus. AB - The effect of improved glycemic control on the prevention or reversal of diabetic nephropathy has been optimally shown by the reduced incidence of albuminuria (accompanied by an increased risk of hypoglycemia) in the Diabetes Control and Complications Trial (DCCT). The earliest detection of the risk or of the presence of diabetic nephropathy continues to be elevated (and confirmed) levels of albuminuria. However, micro- and macroalbuminuria are frequently associated with increased glycated hemoglobin values, complicating attempts to separate the influence of glycemia from an inherent susceptibility for diabetic nephropathy. In the type 1 diabetic patient levels of c-peptide (co-secreted with insulin by the islets of Langerhans) in plasma reflect sustained islet function. C-peptide may be measured in plasma for as long as a decade after the clinical diagnosis of diabetes mellitus, and its presence may be prolonged with better management of diabetes. The consequent improved glycemic control afforded by sustained islet function will reduce the incidence of the retinopathic and nephropathic complications, uniquely accompanied by a lower risk of hypoglycemia. Optimal diabetic management (that is, normal glycemic indices for all diabetic subjects) remains the goal of diabetic therapy. However, failure to normalize glycemic control may remain a reality of contemporary diabetic management. Thus, renal function assays or genetic protocols (each proposed for the earliest detection of diabetic nephropathy) will need to identify individuals at risk against a broadly variable background of glycated hemoglobin levels. PMID- 9407419 TI - Renal structure and function in non-insulin dependent diabetic patients with microalbuminuria. AB - We have recently described heterogeneity in renal structure in non-insulin dependent diabetic patients (NIDDM) with microalbuminuria (MA; defined as albumin excretion rate from 20 to 200 micrograms/min). Thus, at variance with IDDM patients, "typical" diabetic glomerulopathy by light microscopy is observed only in a third of NIDDM with MA (Category II, CII). Further, despite persistent MA, 30% of NIDDM have normal or near normal renal structure (Category I, CI). Another one-third shows "atypical" patterns of renal injury with absent or mild diabetic glomerular changes, associated with disproportionately severe tubulointerstitial lesions and/or arteriolar hyalinosis and global glomerular sclerosis (Category III, CIII). The aims of this study were to evaluate whether similar patterns of renal lesions could be confirmed in a larger group of NIDDM with MA and to investigate tubular function in order to understand the mechanisms underlying MA in NIDDM patients. Renal biopsies were performed in 53 NIDDM with MA. Categories I, II and III were found in 41%, 26% and 33% of NIDDM with MA, respectively. All 8 patients with proliferative diabetic retinopathy were in CII. We also studied the urinary daily excretion rate of alpha 1-microglobulin (alpha 1 m), a low molecular weight protein, which is a useful indicator of tubular function. alpha 1 m was markedly increased only in CII patients (CI vs. CII vs. CIII: 6.2 +/- 1.2 vs. 13.7 +/- 2.1 vs. 7.3 +/- 0.9 mg/day, ANOVA, P < 0.01). In conclusion, we confirm that there is heterogeneity in renal structure in NIDDM patients with MA. This heterogeneity is not due to renal diseases other than diabetes. Increased alpha 1 m and proliferative retinopathy are useful indicators of the subgroup of MA NIDDM patients with typical diabetic glomerulopathy. It is suggested that diabetic microangiopathy explains the simultaneous occurrence of typical diabetic glomerulopathy, proliferative retinopathy and tubular dysfunction in a subgroup of NIDDM patients with MA. PMID- 9407420 TI - Course of renal disease in Pima Indians with non-insulin-dependent diabetes mellitus. AB - The course of renal disease attributable to non-insulin-dependent diabetes mellitus (NIDDM) has been characterized extensively in the Pima Indians of Arizona. Studies in this population indicate that the glomerular filtration rate often increases at the onset of NIDDM and remains elevated as long as normal urinary albumin excretion (< 30 mg albumin/g creatinine) or microalbuminuria (30 299 mg albumin/g creatinine) persist. After the development of macroalbuminuria (> or = 300 mg albumin/g creatinine), the glomerular filtration rate declines at least as rapidly as reported in subjects with insulin-dependent diabetes. Morphologic examination of kidney tissue reveals extensive glomerular sclerosis, mesangial expansion, and widening of epithelial cell foot processes and the glomerular basement membrane in the subjects with macroalbuminuria, but not in those with normo- or microalbuminuria. These findings suggest that substantial structural damage to the kidney occurs at or about the time that macroalbuminuria develops, and the decline in glomerular function in those with macroalbuminuria is due to a loss of ultrafiltration surface area and a reduction in glomerular hydraulic permeability. PMID- 9407421 TI - Kidney function after withdrawal of long-term antihypertensive treatment in diabetic nephropathy. AB - Initiation of antihypertensive treatment in hypertensive non-insulin-dependent diabetic (NIDDM) patients with diabetic nephropathy induces a faster initial (0 to 6 months) and slower subsequent (6 months-end) decline in GFR [delta GFR (ml.min-1.1.73 m-2.month-1) approximately 1.5 vs. 0.4]. Whether this initial phenomenon is reversible (hemodynamic) or irreversible (structural damage) after prolonged antihypertensive treatment is not known. To elucidate these mechanisms we investigated 40 hypertensive NIDDM patients (age 61 +/- 7 years, mean +/- SD), known duration of diabetes 14 years (2 to 33 years) [median (range)] with diabetic nephropathy receiving antihypertensive treatment (angiotensin converting enzyme inhibition, N = 30) for 5 years (1 to 20 years). The following variables were measured the last day on antihypertensive treatment and one month after withdrawal of treatment; GFR (51Cr-EDTA), 24-hour arterial blood pressure (24 hr MABP, Takeda TM2420) and albuminuria (ELISA); the mean 24-hour MABP rose from 102 +/- 11 to 111 +/- 10 (P < 0.0001) and albuminuria [geometric mean (antilog SEM)] increased from 634 (1.3) to 1159 (1.2) (P < 0.0001), while GFR (mean +/- SD) remained unchanged (69 +/- 25 to 70 +/- 26 ml.min-1.1.73 m-2, P = 0.21), after withdrawal of antihypertensive treatment. A significant correlation between the relative change in the 24 hour MABP measurement and the relative change in GFR (r = 0.44, P < 0.01) was found. In conclusion, our results suggest that the faster initial decline in GFR after initiating antihypertensive treatment in hypertensive NIDDM patients with diabetic nephropathy is due to a irreversible effect, and should be accounted for when evaluating the beneficial effect of antihypertensive treatment on the progression of diabetic nephropathy in these patients. PMID- 9407422 TI - Proteinuria predicts end-stage renal failure in non-diabetic chronic nephropathies. The "Gruppo Italiano di Studi Epidemiologici in Nefrologia" (GISEN). AB - We correlated baseline parameters with glomerular filtration rate (GFR) decline and kidney survival in 274 patients with proteinuric non-diabetic chronic nephropathies (creatinine clearance 20 to 70 ml/min/1.73 m2 and proteinuria > 1 g/24 hr over the last three months) enrolled in the Ramipril Efficacy In Nephropathy (REIN) trial. The GFR, evaluated at baseline, one, three and six months after randomization then every six months, declined linearly by 0.52 +/- 0.83 ml/min/1.73 m2/month (mean +/- SD) over a follow-up (median: range) of 21:3 to 52 months, and kidney survival was 64%. In multivariate analysis, higher baseline proteinuria (P = 0.006), and lower GFR (P = 0.0001) and creatinine clearance (P = 0.0001) correlated with a faster GFR decline. Higher proteinuria was the only baseline predictor of a shorter kidney survival (P = 0.0007) and its predictive value was independent of the underlying renal disease, treatment randomization, and blood pressure control during the followup. Patients in the lowest tertile of baseline proteinuria (< 2.5 g/24 hr) had the slowest rate of GFR decline (-0.25 +/- 0.72 ml/min/1.73 m2/month) and the highest kidney survival (94%), compared with patients in the middle tertile (proteinuria 2.5 to 4.3 g/24 hr; delta GFR, -0.59 +/- 0.82 ml/min/1.73 m2/month, P = 0.008; kidney survival 57%, P = 0.0011) and in the highest tertile (proteinuria > 4.3 g/24 hr; delta GFR, -0.79 +/- 0.87 ml/min/1.73 m2/month, P = 0.0001, kidney survival 44%, P = 0.0001). Kidney survival significantly differed even between the middle and highest tertiles (P < 0.05). Thus, in non-diabetic chronic nephropathies proteinuria is an independent and accurate predictor of disease progression and ESRF. PMID- 9407423 TI - Are angiotensin converting enzyme inhibitors superior to beta blockers in retarding progressive renal function decline? AB - We questioned the superiority of angiotensin converting enzyme (ACE) inhibitors to beta blocking drugs with regard to renal function outcome in patients with mild to moderate renal insufficiency and normal to moderately elevated blood pressure (BP). We therefore studied 89 patients in a prospective double-blind randomized trial comparing the effect of enalapril and atenolol on the slope of glomerular filtration rate (GFR). Mean baseline GFR was 53 +/- 20 ml/min, untreated BP 152 +/- 20 mm Hg systolic and 90 +/- 11 mm Hg diastolic and median proteinuria 0.6 g/24 hr (interquartile range 0.0 to 2.5). After a run-in period without antihypertensives, the test drug was titrated to lower diastolic BP to a predefined goal of 10 mm Hg below baseline and/or below 95 mm Hg. The median follow up was 3.9 years. Antihypertensive therapy resulted in a comparable decrease of BP in both study groups. Filtration fraction and proteinuria decreased in both groups. The slope of GFR over time was not different between both groups (-1.39 +/- 2.82 and -1.97 +/- 3.38 ml/min/year on atenolol and enalapril, respectively). In multiple regression analysis a higher baseline GFR, a greater decrease in GFR and in proteinuria during titration and a lower proteinuria during follow up were independently related to a better GFR slope. We conclude that the use of ACE inhibitors is not imperative in all patients with non-diabetic nephropathy. PMID- 9407424 TI - Long-term progression of chronic renal insufficiency in the AIPRI Extension Study. The Angiotensin-Converting-Enzyme Inhibition in Progressive Renal Insufficiency Study Group. AB - The Angiotensin-converting-enzyme Inhibition on Progressive Renal Insufficiency (AIPRI) Study showed that the ACE inhibitor benazepril provides protection against loss of renal function in patients with chronic renal insufficiency (CRI) caused by various renal diseases. As a result of unexpectedly low mortality in the placebo group, there was a substantial imbalance in mortality during the course of this study (8 patients on benazepril vs. 1 on placebo). The aim of the extension study was to follow-up the patients from the AIPRI core study until autumn 1996, focusing on CRI progression and mortality. Data collection was post hoc. Patients were treated according to investigators' usual practices, without knowledge of the core study trial medication or (initially) the core trial results. A new primary efficacy parameter was defined as the time from the start of core study treatment to the occurrence of the first event in the combined composite end-point of dialysis, renal transplantation or death related to renal disease. Serial serum creatinine levels and all-cause mortality were also recorded. The median total follow-up for core + extension periods was 6.6 years. Many patients from both treatment groups (64% on benazepril and 61% on placebo) received ACE inhibitors during follow-up. In the intention-to-treat analysis of the core + extension data, only 79 of 300 patients from the benazepril group, compared to 102 of the 283 patients from the placebo group needed dialysis or renal transplantation, or died related to renal disease (P < 0.013, log-rank test). The mortality imbalance seen in the core trial was not evident with the longer follow-up (25 deaths in the benazepril and 23 in the placebo group, before dialysis). These data clearly demonstrate a long-term beneficial effect in patients randomized to take benazepril during the core study, but because treatment during the extension period was not randomized, the results of this intention-to-treat analysis need to be interpreted with care. PMID- 9407425 TI - Angiotensin II receptor blockade and progression of nondiabetic-mediated renal disease. AB - While the bulk of the existing data are in diabetic renal disease, there are some animal and clinical studies that compare the effects of angiotensin I (AT-1) receptor antagonists to angiotensin converting enzyme (ACE) inhibitors in renal disease of nondiabetic origin. Based on these data, preservation of renal function and morphology occurs with AT-1 receptor antagonists in animal models where renal injury is hemodynamically mediated such as in the remnant kidney. Conversely, in non-hemodynamically mediated renal injury such as in puromycin nephrosis, AT-1 receptor antagonists have not consistently protected against declines in glomerular filtration rate or development of interstitial fibrosis. This may, however, be related to dosage, since high doses of AT-1 receptor antagonists show some protection against progression in these models. It is too early, however, to make judgments regarding the clinical impact of the AT-1 receptor antagonists on progression of nondiabetic renal disease. The result of the ELITE trial support the concept that progression of renal dysfunction associated with heart failure is ameliorated to a similar extent between ACE inhibitors and the AT-1 receptor antagonist, losartan. The AT-1 receptor antagonist group also had fewer side effects including the absence of cough as well as a lower, albeit not statistically significant, incidence of hyperkalemia. Thus, the emerging database supports the concept that AT-1 receptor antagonists have an efficacy similar to ACE inhibitors for preserving renal function and morphology in hemodynamically mediated renal injury. It is unclear, however, whether this drug class will reduce immunologically-mediated renal injury. PMID- 9407426 TI - Clinical correlates to chronic renal allograft rejection. AB - Chronic rejection is a clinical syndrome characterized by a progressive decline in renal allograft function and nonspecific histologic findings of interstitial fibrosis, glomerulosclerosis, and fibrointimal proliferation of intrarenal arteries. Most late allograft failure that is not due to death with a functioning allograft is caused by chronic rejection. Although the pathogenesis and treatment of chronic rejection are unknown, a number of epidemiological studies have examined clinical correlates for possible clues to its pathogenesis. Clinical correlates for chronic renal allograft rejection can be classified in two broad categories: immune (alloantigen-dependent) and non-immune (alloantigen independent). The strongest evidence that chronic rejection is immune mediated comes from its association with acute rejection and the degree of histocompatibility mismatching. However, not all acute rejection leads to chronic rejection, and there is little evidence that newer immunosuppression regimens which effectively prevent acute rejection have reduced the incidence and severity of chronic rejection. Therefore, many clinical investigators have also looked for potential non-immune causes of chronic rejection. Putative non-immune risk factors include donor source (living-related vs. cadaveric), cold ischemia time, delayed graft function, size mismatching, donor age, donor and recipient gender, recipient race, hyperlipidemia, and hypertension. Although there is little evidence supporting a cause and effect relationship between any immune or non immune risk factor and chronic rejection, these clinical associations suggest a number of potentially important areas for further study. PMID- 9407427 TI - Treatment of hypertension in renal allograft patients: does drug selection make a difference? AB - Recent trials have suggested that control of mild to moderate hypertension can slow progression of many forms of chronic renal disease. These findings may apply to renal transplant hypertension. Renal transplant hypertension, however, does not always behave like other forms of hypertension. Thus, clinical trials have not yet shown that blood pressure control will alter the progression of "chronic rejection." What's more, which of the classes of antihypertensive agents might be most effective is also not certain. Most trials suggest that calcium inhibitors and angiotensin-converting enzyme inhibitors have similar effects on blood pressure and glomerular filtration rate in hypertensive transplant patients. PMID- 9407428 TI - Glomerular proteinuria in renal transplant patients: mechanisms and treatment. AB - Proteinuria in chronic kidney transplant failure (CKTF) is due to an alteration of glomerular permselectivity and two major determinants can be characterized experimentally: (1) size selectivity, that is, the ability of the glomerular filter to progressively hinder the passage of macromolecules with increasing molecular radius and (2) charge selectivity, the ability to restrict filtration of negatively charged molecules more effectively than that of equally sized uncharged or cationic compounds. The fractional clearance values of neutral polydisperse dextran molecules create a sieving profile to describe size selectivity, and anionic dextransulfate is used to evaluate charge selectivity. We characterized permselectivity in renal transplants recipients with various degrees of proteinuria. Proteinuria < 1 g/day is caused by an isolated defect of glomerular charge selectivity, whereas nephrotic range proteinuria (characterized histologically by transplant glomerulopathy) is due to an additional impairment of glomerular size selectivity. This sequence is similar to the one observed in patients with chronic native kidney disease (such as diabetic nephropathy). It therefore can be speculated that therapeutic interventions which have been shown to reduce proteinuria in patients with chronic native kidney disease will also beneficially affect permselectivity in CKTF. PMID- 9407429 TI - Impact of new immunosuppressive agents on late graft outcome. AB - Several new immunosuppressive agents have recently been launched for clinical use after kidney transplantation. FK506 or tacrolimus (Prograf) has been tested as an alternative for cyclosporine A (CsA). Both tacrolimus and CsA interfere with the early stage of lymphocyte proliferation by blocking interleukin-2 synthesis. Mycophenolate mofetil (MMF) (Cell-Cept) blocks the de novo synthesis of guanine nucleotides, the pathway essential for purine synthesis in dividing T- and B lymphocytes, and has been used in conjunction with CsA and corticosteroids. In several prospective randomized trials a significant reduction in the incidence of acute rejection has been shown. As early acute rejection and especially recurrent acute rejection, is one of the most important risk factors for chronic rejection and late graft loss, there is a growing interest in the potential beneficial effect of these drugs on the late graft survival. In the European MMF trial, a further reduction of the late graft loss after one year is seen for patients treated with 2 g MMF. A retrospective comparison of international registry data has also shown a significant increase in the graft half-life of patients receiving tacrolimus compared with CsA treated patients. Taking into account the still limited follow-up of the available trials and the fact they were not primarily designed with adequate statistical benefits of these new drugs on the late graft outcome must be made with caution. PMID- 9407430 TI - Antigen-independent determinants of graft survival in living-related kidney transplantation. AB - We used the United Network of Organ Sharing database to define the antigen independent risk factors which contributed to the survival of 8,582 kidney transplants performed in the U.S. between October 1987 and December 1991, using multivariable regression techniques. In this analysis, death with a functioning graft was censored. The risk ratio for graft loss was high when recipients were African-American or had high body surface area, or when donors were older or female. The analysis shows that antigen independent factors that are associated with lower donor kidney mass or increased recipient size play a significant role in living donor kidney transplant loss, as they do in cadaver kidney transplantation. PMID- 9407431 TI - Why is proteinuria such an important risk factor for progression in clinical trials? AB - There are strong reasons to justify the concept that proteinuria is a major risk factor for progression in clinical trials. The evidence is strongest where therapeutic intervention has been focused on established renal disease, when changes in albumin excretion rate (AER) and glomerular filtration rate (GFR) occur within a short time span. Proteinuria is also important in emerging renal disease, such as incipient diabetic nephropathy (DN), since natural history studies show that small increases in AER predict clinical nephropathy and, ultimately, a decline in GFR. However, the absence of concurrent changes in GFR in incipient DN complicates the evaluation of clinical trials in this condition. It is also not certain that the degree of coupling of changes in AER and GFR is the same during intervention as during natural history studies. The importance of proteinuria as a risk factor for progression has been strengthened by recent evidence showing that proteinuria itself causes renal damage. Traditional concepts of the damaging effects of proteinuria have focused on the glomeruli, where mesangial expansion induced by transcapillary passage of proteins has been considered to lead to a decrease in glomerular filtration surface and a decline in GFR. New evidence suggests that interaction of albumin with proximal renal tubules may not only lead to renal damage but may also be causally related to increases in AER. PMID- 9407432 TI - Mechanisms of progression in autosomal dominant polycystic kidney disease. AB - Autosomal dominant polycystic kidney disease (ADPKD) progresses to end-stage renal insufficiency before the age of 73 in approximately 48% of affected individuals. Why the disease, characterized by innumerable cysts arising in proximal and distal tubules, eliminates functioning non-cystic parenchyma in some patients and spares other is a mystery. The cysts initiate in early childhood in fewer than 1% of renal tubules as a consequence of the focal expression of mutated DNA. Tubule cells proliferate, causing segmental dilation, in association with the abnormal deposition of extracellular matrix proteins. Most of the cysts separate from the parent tubules and fill with fluid by cAMP-mediated chloride secretion. Risk factors associated with accelerated loss of renal function include: genotype (PKD Type 1 progresses more rapidly than PKD Type 2); gender (males progress more rapidly than females); race (black patients progress more rapidly than whites); hypertension; proteinuria. The relation between kidney size and progression to renal failure is debated. Progressive PKD is associated with the cellular expression of proto-oncogenes (fos, myc, ras, erb), growth factors (EGF, HGF, acid and basic FGF), chemokines (MCP-1. osteopontin), metalloproteinases, and apoptotic markers, and the interstitial accumulation of Types I and IV collagen, laminin, fibronectin, macrophages and fibroblasts, the magnitudes of which increase with age. Cyst activating factor (CAF), a neutral lipid identified in cyst fluid that stimulates fluid secretion and proliferation of renal epithelial cells and monocyte chemotaxis, has recently been identified as a potential progression factor. In those patients destined to develop renal failure there is loss of non-cystic parenchyma in association with mass replacement by fluid-filled cysts in a network of interstitial fibrosis. The decline in renal function is probably the consequence of processes leading to interstitial fibrosis, as in other nephropathies, rather than due to simple mechanical displacement of parenchyma by cysts. PMID- 9407433 TI - Progression of renal failure: role of apolipoprotein B-containing lipoproteins. AB - Plasma lipoproteins (LP) may be identified on the basis of density properties or apolipoprotein (apo) composition. ApoB-containing LP occur in VLDL, IDL and LDL. There are several types of apoB-containing LP characterized by specific composition of minor apolipoproteins (apoC, apoE etc.) and lipid constituents (triglycerides and cholesterol), metabolic properties and relative atherogenicity. The alterations of lipoprotein metabolism in renal disease resulting in elevated levels of apoB-containing LP may be reflected in hyperlipidemia. Whereas nephrotic syndrome and heavy proteinuria are associated with increased formation of cholesterol-rich apoB-containing LP in LDL and VLDL, the characteristic feature in renal failure is the accumulation of intact or partially metabolised triglyceride-rich LP in IDL and VLDL. The potentially atherogenic apoB-containing LP have been linked to the pathogenic processes that result in progressive glomerular and interstitial lesions and ultimate loss of renal function. The mechanisms of injury are not fully understood. Receptor- and non-receptor mediated uptake of LP by mesangial cells may induce or accelerate proliferative and sclerotic processes in the glomerular mesangium that are analogous to atherosclerosis in the arterial wall. Changes in glomerular permeability can result in increased filtration of LP that may be internalized by tubular cells and elicit corresponding lesions in the interstitial tissues. The negative impact of proteinuria on the prognosis of renal disease could be mediated in part through an increased filtration of lipoproteins. Induction of hyperlipidemia accelerates glomerular and interstitial damage in experimental renal failure. This can be attenuated by treatment with hypolipemic agents. In patients, increased concentrations of apoB-containing LP are associated with more rapid progression of renal insufficiency in both primary renal disease and diabetic nephropathy. It is, however, presently not known to what extent treatment of the renal dyslipidemia can modify the progression of chronic renal failure. Experimental and clinical evidence suggest that apoB-containing LP may play a pathogenetic role in the progression of renal disease. PMID- 9407434 TI - Lipids and progression of renal disease: role of modified low density lipoprotein and lipoprotein(a). AB - Atherogenic lipoproteins accumulate in the arterial wall as well as within the glomerulus and may accelerate vascular and glomerular injury. We therefore assessed whether oxidized low density lipoprotein (LDL) and lipoprotein(a) [Lp(a)] influence three major systems: (i) endothelium-dependent vasodilation, (ii) renin release of juxtaglomerular (JG) cells, and (iii) proliferation and viability of mesangial cells (MC). Lipoproteins were prepared from human plasma. Renal arteries were obtained from rabbits and JG as well as MC cells from mouse, rat and human kidneys. Dilator responses were detected in isolated arterial segments by a photoelectric device. Renin activity of JG cells was measured in culture supernatants and cells and DNA synthesis by 3H-thymidine incorporation in MC. Acetylcholine-induced, endothelium-dependent dilator responses of renal arteries were not significantly attenuated after incubation with native Lp(a). However, exposure to in vitro oxidized Lp(a) suppressed dilator responses in a dose-dependent manner. Using a chemiluminescence assay, we could detect increased O2- production by arteries pretreated with oxidized Lp(a), which suggested that enhanced nitric oxide (NO) inactivation by O2- might be the underlying mechanism of impairment of endothelium-dependent dilations. In general, oxidized Lp(a) was far more potent than oxidized LDL in this effect. In JG cells, both oxidized LDL and Lp(a) dose-dependently stimulated renin release. Coincubation with HDL significantly suppressed oxidized LDL and Lp(a) stimulated renin release and O2- production. In MC native and oxidized Lp(a) were poor ligands for the LDL receptor, but bound more tightly to extracellular matrix than native LDL. Native and oxidized Lp(a) elicited proliferation or toxicity of MC in a dose-dependent fashion. Stimulation of DNA synthesis in MC or renin release in JG cells was partly blunted or eliminated when cells were incubated with oxidized LDL and Lp(a) in the presence of superoxide dismutase and catalase, enzymes removing O2- and H2O2. These dat suggest a common underlying mechanism. Atherogenic lipoproteins induce formation of oxygen radicals not only in arteries, but also in glomeruli and JG cells, causing an inhibition of nitric oxide mediated vasodilation, stimulation of renin release, and modulation of mesangial cell growth and proliferation. The damaging effect of the lipoproteins can be prevented by antioxidative enzymes and HDL. PMID- 9407435 TI - Role of local and systemic angiotensin in diabetic renal disease. AB - Classically, the renin-angiotensin system (RAS) in diabetes was thought to be suppressed, and relatively unimportant in the regulation of hemodynamics and the development of complications. However, studies of pharmacologic interruption of the RAS with angiotensin converting enzyme (ACE) inhibition have implicated the RAS in the progression of diabetic nephropathy. Preliminary evidence also suggests a beneficial effect of angiotensin II receptor antagonists. The relative roles of the systemic versus intrarenal RAS in this process are under active investigation. Though plasma renin is generally low, there may be subtle changes in angiotensin (Ang) II metabolism that sustain relatively higher plasma Ang II levels. Furthermore, the intrarenal RAS may not be suppressed. Renal renin levels tend to be disproportionately elevated, as compared to plasma values. Renal Ang II levels are normal, and renal mRNAs for RAS components have been variable. In general, lack of intrarenal RAS suppression (despite plasma volume and increased exchangeable sodium) may indicate inappropriate activity of the local tissue RAS, and act as a proximate cause of the systemic RAS suppression. Ang II-mediated injury may occur via stimulation of sclerosing mediators, and there is evidence that hyperglycemia acts synergistically with Ang II to promote cellular injury. Together, these recent investigations lend further support to the notion that the RAS plays an important role in diabetic nephropathy, and are helping to shed light on the mechanisms of progressive renal injury. PMID- 9407437 TI - Aldosterone is a major factor in the progression of renal disease. AB - There is compelling evidence supporting the renin-angiotensin-aldosterone system contribution in experimental and human renal disease. Interruption of this system by converting enzyme inhibition or angiotensin II receptor antagonism reduces injury. Angiotensin II contributes to the progression of renal disease through its direct vascular effects and proliferative properties. The mediators of angiotensin II induced renal injury are many and include TGF-beta, PDGF, bFGF, and endothelin. Though the mechanisms involved in its contribution to progressive renal disease are not well delineated, aldosterone seems to be an overlooked contributor to the progression of kidney disease and its effects may also depend on both its hemodynamic and more direct cellular actions. PMID- 9407436 TI - Comparative study of ACE inhibitors and angiotensin II receptor antagonists in interstitial scarring. AB - Many of the pathophysiologic events associated with kidney disease are driven by angiotensin II. Irrespective of the etiology, many kidney diseases lead to tubulointerstitial inflammation, fibrosis and loss of renal function. Contributors to the process of tubulointerstitial fibrosis include monocyte/macrophage infiltration, the synthesis of profibrotic cytokines such as transforming growth factor beta 1 (TGF-beta 1), interstitial myofibroblast proliferation, and clusterin expression. These processes are ameliorated by angiotensin converting enzyme (ACE) inhibition. Blockade of the angiotensin II receptor (AT-1) impaired fibroblast proliferation, consequent differentiation into myofibroblasts, and the synthesis of TGF-beta 1, but did not prevent monocyte infiltration. TGF-beta 1 synthesis or fibroblast proliferation but prevented the differentiation of fibroblasts into myofibroblasts and blocked clusterin expression. The nuclear factor-kappa B (NF-kappa B) family of transcription factors regulates genes involved in inflammation, proliferation and differentiation. ACE inhibition, AT-1 and AT-2 receptor blockade each differentially attenuated NF-kappa B isotype activation. The changes in NF-kappa B isotype may account for the variation seen in the pharmacologic effect of angiotensin II formation or action on the fibrotic process. When considering therapeutic options to prevent renal disease progression, one must be aware of the impact of transcription factors on the injured kidney and the consequent changes in cell infiltration, proliferation and differentiation. PMID- 9407438 TI - Glomerulosclerosis, arteriosclerosis, and vascular graft stenosis: treatment with oral heparinoids. AB - At present there is no known treatment for established glomerulosclerosis or atherosclerosis. Since the principal lesion in glomerulosclerosis involves mesangial cells, a vascular smooth muscle cell, we searched for new therapeutic approaches affecting vascular smooth muscle function, especially with respect to modifying the turnover of extracellular matrix. We used mice transgenic for bovine growth hormone (bGH), since these mice develop end-stage renal disease due to progressive glomerulosclerosis. We previously showed that the subcutaneous injection of a non-anticoagulant heparin reduced glomerulosclerosis in bGH mice. Since injectable drugs are not a practical means of controlling glomerulosclerosis in humans, we assessed oral heparin-like compounds and found that oral pentosan polysulfate (PPS) reduced glomerulosclerosis in bGH mice at non-toxic doses. Because the positive therapeutic response in the bGH model could have been principally hormone-mediated, we examined other models of non-immune mediated glomerulosclerosis, including ROP Os/+ non-diabetic and diabetic mice. We found that an oral PPS (Elmiron), which is approved for other indications in humans, reduced sclerosis in all of these forms of chronic, progressive glomerulosclerosis. Based on the similarity of the cellular events in glomerulosclerosis and arteriosclerosis, we assessed the effect(s) of PPS in congenital (Watanabe rabbits) and induced (New Zealand White lipid-fed rabbits) models of arteriosclerosis. The extent and severity of the lesions was significantly reduced in both models by PPS treatment. Finally, we asked whether the proliferative and sclerotic lesion, which is the cause of vascular graft stenosis, might also respond to PPS treatment. To do this we cultured cells from the materials removed from stenotic arteriovenous grafts in hemodialysis patients. We found that PPS inhibits the proliferation and matrix production in a dose-dependent manner. PMID- 9407439 TI - Nephron mass as a risk factor for progression of renal disease. AB - Partial ablation of renal mass initiates a cycle of progressive glomerular injury in the remnant. This process is associated with glomerular hypertrophy, hyperfiltration and systemic hypertension. Congenital deficits in nephron number are also associated with adverse effects on the kidney. Since intrauterine growth retardation is associated with formation of fewer nephrons, the recent observation that low birth weight is associated with increased risk of hypertension in later life raises the possibility that even modest reductions in nephron complement may also predispose to renal injury. Likewise, the numbers of viable nephrons supplied to renal, allograft recipients may be critical determinants of late allograft success or failure, since subsequent acute ischemia and rejection combine to lower the nephron complement to levels akin to the more extensive reductions in renal mass seen in patients with surgical reduction of a solitary kidney, in whom predisposition to hypertension and glomerulosclerosis is evident. This article summarizes recent findings suggesting that congenital nephron endowment is a significant factor in the pathogenesis of chronic renal disease and hypertension. PMID- 9407440 TI - Heterogeneity of angiotensin action in renal circulation. AB - Reported concentrations of angiotensin II (Ang II) in renal interstitial fluid are as high as 10 nM. Despite such high concentrations, intrarenal arterial infusion of Ang II at rates that induce far less change in renal Ang II concentration still elicits renal vasoconstriction. We examined whether the glomerular afferent arterioles (Af-Art) was more sensitive to intraluminal than extraluminal Ang II in superficial or juxtamedullary nephrons. Rat superficial Af Arts with the intact glomerulus were microdissected and perfused in vitro at 70 mmHg, while juxtamedullary Af-Arts were visualized in isolated perfused kidneys (at 100 mm Hg) according to the method of Casellas and Navar. Increasing doses of Ang II (1 pM to 10 nM) or norepinephrine (NE; 1 nM to 1 microM) were added to either bath (extraluminal) or arteriolar perfusate (intraluminal). Decreases in luminal diameter induced by Ang II were significantly larger with intraluminal than extraluminal administration in superficial Af-Art: at 100 pM the diameter decreased by 52 +/- 8% (N = 6) and 7 +/- 3% with intraluminal and extraluminal administration, respectively. In contrast, in the juxtamedullary Af-Arts intraluminal and extraluminal Ang II caused similar constriction. On the other hand, there was no difference in intraluminal versus extraluminal NE action in either superficial or juxtamedullary nephrons. In conclusion, glomerular Af-Arts seem to have a higher sensitivity to luminal than interstitial Ang II in superficial but not juxtamedullary nephrons. Such heterogeneities in Ang II action may be important in the control of glomerular hemodynamics under various physiological and pathological conditions. PMID- 9407441 TI - Role of angiotensin II generated by angiotensin converting enzyme-independent pathways in canine kidney. AB - Recent studies have provided evidence of angiotensin converting enzyme (ACE) independent angiotensin (Ang) II formation in tissue renin-angiotensin systems. We studied the effects of Ang II generated by ACE-independent pathways on renal hemodynamics. We used a synthetic peptide, [Pro11, D-Ala12]-Ang I (S), which yields Ang II by chymase, but not by ACE. Infusion of Ang I into a renal artery caused a decrease in renal blood flow, and reciprocally an increase in mean arterial pressure. Infusion of S (1 nmol/kg) caused a decrease in renal blood flow (-20%), but a larger dose was needed to increase mean arterial pressure. Studies with an intravital needle-probe CCD camera revealed that the Ang I infusion induced dose-dependent vasoconstriction of afferent and efferent arterioles (49% and 54%, respectively at 1 nmol/kg). In contrast, infusion of S elicited only 30% constriction of these vessels at a dose of 1 nmol/kg and induced no further constriction at higher doses, indicating that different segments of renal vessels responded in different fashions to Ang II formed via ACE-independent pathways. These vasoconstrictions were abolished by an angiotensin II receptor (AT-1) antagonist. Enzymatic assays using reverse-phase HPLC revealed that the ACE-dependent pathway was predominant in the rena1 cortex (approximately 80%). We also determined Ang II concentrations in renal cortex specimens obtained by needle biopsy. Intrarenal S infusion (10 nmol/kg) increased plasma and renal Ang II concentrations to 160% and 710% of the respective baseline levels. This study provides in vivo evidence of ACE-independent Ang II formation in renal tissue and suggests that this locally-formed Ang II influences the renal circulation in a paracrine fashion. PMID- 9407442 TI - Effects of candesartan cilexetil (TCV-116) and enalapril in 5/6 nephrectomized rats. AB - The renal protective properties of candesartan cilexetil (TCV-116), an angiotensin II type 1 receptor antagonist (AT1A), and enalapril, an angiotensin I converting enzyme inhibitor (ACEI), were investigated in 5/6 nephrectomized (NX) rats. Candesartan cilexetil (1 mg/kg/day) and enalapril (10 mg/kg/day) were administered orally to 5/6 NX rats for four weeks (during the early phase of disease development) or 16 weeks (through the late phase). In vehicle-treated rats, proteinuria, glomerulosclerosis, interstitial mononuclear cell (MNC) infiltration and interstitial fibrosis developed. Moreover, immunohistological studies showed enhanced expression of transforming growth factor-beta 1 (TGF-beta 1) in the injured glomeruli. Both drugs inhibited these adverse changes in the early phase. In the late phase, the progressive proteinuria, interstitial MNC infiltration were attenuated by both drugs. However, candesartan cilexetil significantly inhibited the progression of glomerulosclerosis, the expression of TGF-beta 1 and the interstitial fibrosis, while enalapril did not. Candesartan cilexetil and enalapril showed comparable hypotensive effects after the 16-week administration. These results indicate that candesartan cilexetil shows a more potent protective effect than enalapril against the progression of renal injury in the late phase. Thus, an AT1A might be more useful than an ACEI for the treatment of patients with chronic renal failure. PMID- 9407443 TI - Angiotensin receptor antagonists in experimental models of chronic renal failure. AB - The efficacy of angiotensin converting enzyme inhibitors (ACEI) in slowing the advancement of chronic renal disease attests to the importance of angiotensin II (Ang II) in the pathophysiological mechanisms underlying disease progression. It is apparent from studies of the effects of orally-active AT1 receptor antagonists (AT1RA) in experimental models of chronic progressive renal disease, that AT1RA have broadly similar effects to those of ACEI, implying that the favorable effects of ACEI on systemic and renal hemodynamics and indices of glomerular injury are mediated, in large part, by reducing the action of Ang II at AT1 receptors. The possibility remains, however, that differences in the modes of action of ACEI and AT1RA are significant in terms of renal protection and that the two classes of drugs are not therapeutically equivalent. Thus far, however, virtually all experimental studies comparing the renal protective effects of ACEI versus AT1RA have failed to show any convincing differences between the two classes of drug that cannot be attributed to discrepancies in the levels of blood pressure control achieved. As many rodent studies have adopted protocols originally designed to distinguish between the effects of treatment versus no treatment, however, it may be premature to conclude that ACEI and AT1RA are, essentially, therapeutically equivalent. Since both classes of drug have such potent renoprotective effects, the extent of injury that develops in treated rats may be so slight as to compromise the sensitivity of the experimental comparison. Fresh experimental approaches may be required to overcome this issue and resolve any outstanding questions concerning the therapeutic equivalence of AT1RA and ACEI in slowing the progression of renal disease. PMID- 9407444 TI - Assessment of structure and function in progressive renal disease. AB - The incidence and prevalence rates of end-stage renal disease (ESRD) in the United States continue to increase. In 1995, the incidence rate was 262 per million population, with a point prevalence rate of 975 per million population. The exact number of individuals with abnormal renal function but not yet at end stage is difficult to assess. Crude estimates suggest that approximately 0.4% of the U.S. population has serum creatinine values greater than 2.0 mg/dl. In some sub-populations, such as African Americans, the estimate is + as high as 1.0%. The rate of progression, likewise, is difficult to assess. In general, renal manifestations of certain systemic diseases such as diabetes mellitus and systemic lupus erythematosus, and those with significant proteinuria (usually greater that 3.0 g/24 hr) seem to have a more rapid progressive course to end stage. If intervention is expected to be successful in halting or slowing down progression, accurate assessment of the early manifestations of renal disease, structure, and function need to be established. Currently accepted methods of assessment of renal disease include measurement of renal function such as serum creatinine and glomerular filtration rate, measurement of proteinuria, assessment of tubular function, glomerular sieving and permselectivity, radiologic imaging techniques, and evaluation of histo-morphometry. Interventions that have been shown to slow progression include control of hypertension, and treatment modalities that reduce proteinuria, such as, the use of angiotensin converting enzyme inhibitors. Further clinical and basic science studies are needed to accurately define the important predictors of progression, and interventions that are effective in slowing or halting progression. PMID- 9407445 TI - Measurement of glomerular filtration rate. AB - Glomerular filtration rate (GFR) is the standard measure of renal function and is critical for the diagnosis and management of renal diseases. Rigorous assessment of GFR requires the measurement of renal clearance of an exogenous marker that is freely filtered by the kidney, and that does not undergo metabolism, tubular secretion or absorption, such as inulin. While its clearance provides the most accurate method of measuring GFR, it is not suitable for routine clinical practice. Labeled compounds as alternative filtration markers, including 125I iothalamate and 99mTc-diethylenetriaminepenta-acetic acid (DTPA), provide accurate and precise GFR measurements, but their use may be limited for safety reasons. To avoid exposing patients to radiation, investigators have proposed clearance procedures using minute doses of non-radioactive contrast agents, including iothalamate (ionic) and iohexol (non-ionic). This approach provides similar accuracy to inulin clearance. The most important limitation of all renal clearance methods is that urine is collected by spontaneous voiding, stimulated by water loading, and thus subject to errors due to incomplete emptying of the bladder. Thus, plasma clearance of a suitable exogenous marker (51Cr-EDTA, 125I iothalamate, iohexol) has been suggested for measuring renal function, in which the elimination rate of the tracer after a single intravenous injection is evaluated. Plasma clearance of these markers estimated by multiple blood samples provides more precise information, but repeated sampling makes this method cumbersome. To overcome this drawback, abbreviated kinetic profiles have been proposed to predict GFR from the plasma disappearance curve (elimination phase). On analyzing the data with a simplified method that uses a one-compartment model (six blood samples only in the elimination phase), corrected with the Brochner Mortensen formula, an excellent correlation was found with inulin clearance. This method is currently employed for measuring GFR in multicenter clinical trials. PMID- 9407446 TI - Diabetic nephropathy as a model for the use of renal structural endpoints in clinical trials. AB - Slowly developing renal diseases (SDRD) such as diabetic nephropathy (DN) and hypertensive nephrosclerosis constitute the large majority of disorders leading to end-stage renal disease (ESRD). These disorders are characterized by years to decades without renal functional abnormalities during which renal structural changes are developing. The earliest renal functional abnormalities [such as microalbuminuria in patients with insulin dependent diabetes mellitus (IDDM)] are often associated with already well established renal lesions that may progress independent of the initiating cause. Since the clinical manifestations of these disorders are dependent upon established structural changes, prevention of the early lesions seems a logical goal and thus a useful endpoint for clinical trials. Functional endpoints are impractical at these early stages as they would take too long to manifest. Diabetic nephropathy in IDDM patients is a useful model for such study design, since there is sufficient knowledge of the natural history of the disorder and of the important structural endpoints to allow for the statistical power calculations crucial for study design. More information regarding natural history and structural-functional relationships is needed in non-insulin dependent diabetes, hypertension, and other SDRD before intervention trials using renal structural endpoints can be developed for these important causes of ESRD. PMID- 9407447 TI - Economic evaluation of benazepril in chronic renal insufficiency. AB - A prospective, randomized, double-blind trial recently demonstrated that treating patients with chronic renal insufficiency with benazepril significantly decelerates the rate of progression of the disease. We tested the hypothesis that preventative treatment with the angiotensin converting enzyme (ACE) inhibitor benazepril in patients with chronic renal insufficiency is cost-effective. A Markov chain model was used that considered regular treatment, hemodialysis, continuous ambulant peritoneal dialysis, transplantation, rejection and death. Clinical trial data were used to estimate the effects of benazepril treatment and to estimate the duration until renal replacement therapy was needed. Epidemiologic parameters were derived on the basis of Dutch registries of renal diseases, costs are estimated by updating former estimates, literature review and expert opinion. We found that preventative treatment with benazepril decreased the percentage of patients who died or developed end-stage renal disease. Total costs per patient are expected to decrease in three years with more than $4,000 US per patient. Extrapolated to ten years, the savings are estimated at $23,500 US per patient. Benazepril treatment is not only an effective treatment in patients with chronic renal failure. By increasing the years spent without dialysis, it is also a cost-effective treatment. PMID- 9407448 TI - Microalbuminuria: a marker of cardiovascular risk and organ damage in essential hypertension. AB - Microalbuminuria (Mi) is thought to reflect diffuse vascular damage and to predict cardiovascular morbidity and mortality in essential hypertension, although its pathogenesis remains to be fully elucidated. The relationship between microalbuminuria and several cardiovascular risk factors and target organ damage was evaluated in a large cohort of untreated essential hypertensive patients. Albuminuria was measured as the albumin to creatinine ratio in three non consecutive first morning urine samples. Cardiac damage was evaluated by ECG and retinal vascular changes by direct ophtalmoscopy. In a subgroup of 23 patients with Mi and in a control group of 22 normoalbuminurics, selected from the entire cohort of patients and carefully matched for age, gender, body mass index (BMI) and duration of disease, we also measured left ventricular mass index by M-B mode echocardiography, common carotid wall thickness by high resolution US scan, and renal vascular resistances by US-doppler of interlobar arteries. K means cluster analysis performed on the entire cohort of patients showed that microalbuminuria is associated with the presence of an unfavorable risk profile and target organ damage. Furthermore, microalbuminuric hypertensive patients have a larger left ventricular mass index, increased intima media thickness of carotid arteries and higher intrarenal vascular resistances as compared to a well matched group of normoalbuminuric patients. We conclude that in essential hypertension increased urinary albumin excretion can be useful to identify patients for whom more aggressive preventive strategies and/or additional treatment measures are advisable. PMID- 9407449 TI - Creatinine clearance predicted from body cell mass is a good indicator of renal function. AB - The aim of this study was to evaluate the agreement between the glomerular filtration rate (GFR) and an estimate of creatinine clearance (CCr), calculated from the values of body cell mass (BCM) and of plasma creatinine (PCr), thus avoiding urine collection. The value of BCM was obtained from the measurement of total body impedance in 80 renal patients. The relationship between 24-hour urinary creatinine excretion and BCM was evaluated in 30 of these patients. Body cell mass CCr (ml/min) was then calculated in each of the remaining 50 patients. For comparison, 24-hour CCr was measured in the same patients. The correlation coefficient of BCM CCr with GFR was 0.961, while that of 24-hour CCr with GFR was 0.869. Also, the agreement with GFR was better for BCM CCr than for 24-hour CCr. In conclusion, creatinine clearance predicted from body cell mass and plasma creatinine is a better indicator of renal function than measured 24-hour creatinine clearance. PMID- 9407450 TI - Predictors of end-stage renal disease and body mass index in a screened cohort. AB - The effect of body mass index (BMI) on the risk of developing end-stage renal disease (ESRD) has not been critically evaluated. To investigate this issue we used the registries of community-based mass screening and chronic dialysis programs. In 1983, a total of 101,516 subjects (47,901 men and 53,615 women) participated in a mass screening program in Okinawa, Japan. A total of 187 (101 men and 86 women) entered an ESRD program by March 31, 1994. Body mass index was calculated as weight in kilograms divided by the square of the height in meters. Subjects were categorized into six groups based on BMI: < 20.0, 20.0-21.9, 22.0 23.9, 24.0-25.9, 26.0-27.9, and > or = 28.0 kg/m2. The cumulative incidence of ESRD, per 100,000 subjects was calculated in each BMI subgroup for men and women. In men, the cumulative incidence of ESRD increased with BMI, except in the range of 24.0 to 25.9 kg/m2. In women, the association between the cumulative incidence of ESRD and BMI was not clear, but was lowest in the range of 24.0 to 25.9 kg/m2. The adjusted odds ratio (95% confidence interval) was 0.99 (0.92 to 1.13) in men and 0.83 (0.72 to 0.96) in women. PMID- 9407451 TI - Podocyte detachment and epithelial cell reaction in focal segmental glomerulosclerosis with cellular variants. AB - In primary focal segmental glomerulosclerosis with cellular variants the type of epithelial cells that segmentally or globally increased and their relation to sites where podocytes detached from the capillary wall, remain unclear. Two renal samples containing glomeruli with cellular variants were serially examined using transmission electron microscopy. A total of 10 lesions, including 2 collapsing glomerulopathy, 7 cellular lesions, and 1 sclerosis, were examined. Disappearance of the dense basal band of microfilaments from portions of podocytes was followed by detachment from the capillary basement membrane, resulting in segmental denudation of the capillary wall. The defects were not covered by other portions of epithelial cells, when the defects occurred in more central regions of the glomerulus. When detachment of podocytes occurred in the peripheral areas of the glomerulus, the defects were covered with new basement membrane and by other epithelial cells. These epithelial cells were characterized by the formation of intercellular junctional complexes of zonula adherens and desmosomes between each other and with the parietal epithelial cells (PECs), by showing occasional cilia, apoptotic nuclei, and mitosis. Thus, repair is achieved as the defect is covered by the participation of PECs; synechia and glomerular sclerosis result, depending upon the extent of the original podocyte injury. PMID- 9407452 TI - Prognosis of Japanese membranous nephropathy patients with nephrotic syndrome. AB - We retrospectively investigated the prognosis and efficacy of various treatments in 58 Japanese patients with primary membranous nephropathy (MN). Patients were treated with prednisolone (PSL) + immunosuppressive (IS) agents (N = 22). PSL alone (N = 23), IS agents alone (N = 5) and aspirin or dipyridamole (N = 8). Overall, 25 patients (43.1%) achieved complete remission (CR), 11 (18.9%) achieved partial remission (PR) and 22 (37.9%) were non-responders (NR). The 10 year survival rate was 90% and the 10-year CR rate was 61.8% as determined by Kaplan-Meier analysis. There was no significant difference in the CR among treatment groups. Of the 22 patients in the PSL + IS group, 3 progressed to end stage renal disease and 2, who were more than 65 years old, died of pneumonia. The clinical and histological data at renal biopsy did not significantly differ among the treatment groups. In conclusion, the overall prognosis of MN was excellent. Treatment with PSL and/or IS agents did not appear to be superior to treatment with aspirin or dipyridamole. PMID- 9407453 TI - Immunohistochemical localization of latent transforming growth factor-beta binding protein in IgA nephropathy. AB - It has been demonstrated that cultured mesangial cells (MC) express latent transforming growth factor (TGF)-beta binding protein (LTBP) mRNA, and that LTBP might be essential for the extracellular matrix (ECM) accumulation stimulated by TGF-beta in cultured MC. We performed a study to test the pathophysiological role of LTBP in mesangial ECM accumulation in human glomerulonephropathies. The expression of LTBP in 64 renal biopsies of patients with renal diseases was examined by immunohistochemical staining with the specific antibody raised against human LTBP. The biopsy specimens were divided into three groups: Group 1, IgA nephropathy; Group 2, IgA negative mesangial proliferative glomerulonephritis, which mainly consisted of diabetic nephropathy and lupus nephritis; and Group 3, non-proliferative nephropathy, which consisted of minimal change and membranous nephropathy. Immunohistochemical staining of LTBP was significantly detected in mesangial/paramesangial area of glomerulus in Group 1, but not observed in Group 2 or Group 3. The intensity of staining was well related to the grade of mesangial ECM accumulation in Group 1. These findings suggest that the LTBP-TGF-beta complex may play a pivotal role in ECM accumulation in IgA nephropathy, and that modification of LTBP-ECM interaction might provide a new therapeutic strategy against the progression of glomerulosclerosis. PMID- 9407454 TI - Renal hemodynamic effects of candesartan in normal and impaired renal function in humans. AB - The effects of angiotensin II type I receptor antagonist candesartan cilexitil, 8 mg once daily, were studied after single dose and after five days treatment in 17 hypertensive patients [median mean arterial pressure (MAP) 118 mm Hg, range 84 to 134] with renal function impairment of different severity [glomerular filtration rate (GFR) 60 ml/min, range 11 to 161]. The MAP fell by -8% (-14 to -5) and -11 ( 16 to -5)% after single and multiple dose, respectively (both P < 0.02). Effective renal plasma flow (ERPF) increased by 13% (7 to 19) and 10% (3 to 14) after single and multiple dose, respectively (both P < 0.02), while the GFR did not change. Filtration fraction (FF) fell by -11% (-14 to -5) and -12% (-13 to 4) after single and multiple doses, respectively (both P < 0.02). After a single dose the % change in ERPF (r = 0.58) and FF (r = -0.52, both P < 0.05) positively correlated with pretreatment GFR, indicating a more pronounced response in patients with normal GFR. After five days of treatment these correlations were absent, indicating similar renal vasodilation in patients with normal and impaired renal function. Thus, multiple dose candesartan cilexitil had a favorable renal hemodynamic profile, irrespective of pretreatment renal function. Further studies are needed to establish whether this provides long-term renoprotection as well. PMID- 9407455 TI - Effects of oral adsorbent AST-120 on the progression of chronic renal failure: a randomized controlled study. AB - This prospective, randomized controlled study was designed to examine the effects of oral adsorbent AST-120 on the progression of chronic renal failure (CRF) in patients on a strict low protein diet (LPD). Twenty-six patients with CRF (serum creatinine 3.0 to 8.6 mg/dl) on a LPD were randomly assigned to a control group (N = 13) or an AST-120 group (N = 13). The 1/Cr slope and creatinine clearance (CCr) slope were used to estimate the progression rate of CRF; uremic toxins, serum and urinary indoxyl sulfate (IS), peak 2a and guanidino substrates (GS) measured by HPLC. Comparisons were made between the baseline observation period for 6 to 12 months and the treatment period (0.6 g/kg/day of LPD alone or concurrent with 6 g/day of AST-120, for the control and the AST-120 groups, respectively) for 12 to 24 months in both groups. Both the 1/Cr slope and CCr slope were significantly lessened in the treatment period only in the AST-120 group. Serum and urinary IS, but neither peak 2a nor GS were significantly decreased in the treatment period only in the AST-120 group. We conclude that AST 120 administration concurrent with LPD may be superior to LPD alone in retarding the progression of CRF by inhibiting accumulation of indoxyl sulfate. PMID- 9407456 TI - Tubulointerstitial lesions in non-insulin dependent diabetes mellitus. AB - The clinical features and quantitative renal morphometry, including the index of mesangial expansion (IME), the index of arteriolar hyalinous change (IAHC), the percentage of globally sclerosed glomeruli (%GS), and interstitial volume fraction for total renal cortex (Vv int/T) of 67 Japanese non-insulin dependent diabetes mellitus (NIDDM) patients were examined. For the total subject population, Vv int/T correlated with urine protein, creatinine clearance, blood pressure, IME, IAHC and %GS, but not with duration of NIDDM. No significant difference in interstitial expansion between normo- and microalbuminuric patients was observed, and Vv int/T did not correlate with urine albumin excretion rate in those patients. However, in the area without hyalinized glomeruli and atrophic tubules, we found significant interstitial expansion and thickening of the tubular basement membrane (TBM) in patients without overt proteinuria as compared with those in the age-matched minimal change control group. This study confirmed that interstitial expansion in NIDDM progresses in parallel with glomerular and arteriolar lesions and that the expansion occurs concurrently with TBM thickening before the appearance of proteinuria. PMID- 9407457 TI - Urinary excretion of TGF-beta 1, PDGF-BB and fibronectin in insulin-dependent diabetes mellitus patients. AB - We studied the urinary excretion of transforming growth factor-beta 1 (TGF-beta 1), platelet-derived growth factor-BB (PDGF) and fibronectin (FN) in 104 patients (52 normoalbuminuric, 24 microalbuminuric, and 28 with overt diabetic nephropathy) with insulin-dependent diabetes mellitus (IDDM) of a long duration and in 30 non-diabetic controls. IDDM patients had higher urinary excretion of TGF-beta 1, PDGF and FN compared to controls. Urinary excretion of TGF-beta 1 and PDGF was elevated in all IDDM subgroups, while FN excretion was significantly increased only in patients with macroalbuminuria. Urinary excretion of TGF-beta 1 and FN did not differ between normoalbuminuric IDDM patients with long duration of diabetes, a group at low risk of ever developing diabetic nephropathy, and IDDM patients with incipient or overt diabetic nephropathy. Excretion of PDGF was significantly higher in patients with micro- and macroalbuminuria compared to normoalbuminuric patients, but there was a considerable overlap between the groups. In conclusion, although longitudinal follow-up studies are needed to further clarify the issue, our results in long-standing IDDM do not support a hypothesis of urinary excretion of TGF-beta 1, PDGF or FN to predict development of diabetic nephropathy. PMID- 9407458 TI - Mechanism of decreased albuminuria caused by angiotensin converting enzyme inhibitor in early diabetic nephropathy. AB - The mechanism of decreased albuminuria caused by an inhibitor of angiotensin converting enzyme (ACE) was investigated in patients with early diabetic nephropathy. The subjects were 10 patients with non-insulin-dependent diabetes mellitus without azotemia but with albuminuria (less than 650 mg/day). First, a two-week study was done: one week with a diet with ordinary sodium levels and one week with a sodium-restricted diet, in random order. The systemic blood pressure and urinary excretion of sodium and albumin were measured daily. Intrarenal hemodynamics, in terms of the resistance of afferent and efferent arterioles (RA and RE) and glomerular capillary pressure (PGC), were calculated from renal clearance, the plasma total protein concentration, and the pressure-natriuresis relationship. Results obtained before and two weeks after starting the ACE inhibitor cilazapril (2 mg/day) were compared. Urinary excretion of albumin was decreased by cilazapril in 8 of the 10 patients. Cilazapril decreased the RE [6830 (3680, 14,750) to 4660 (1750, 10,790) dynes.sec.cm-5, P < 0.05, mean (minimum, maximum)] and PGC (53 +/- 5 to 43 +/- 9 mm Hg, P < 0.02, mean +/- SD) in these 8 patients, but not in the two other patients. The RA was not significantly changed in any patient. The percent change caused by cilazapril in the urinary excretion of albumin was significantly correlated with the change in PGC (N = 10, r = 0.875, P < 0.01), but not with changes in the systemic blood pressure. In conclusion, the mechanism by which an ACE inhibitor caused a short term decrease in albuminuria in early diabetic nephropathy involved a glomerular hemodynamic change, namely, a decrease in PGC. PMID- 9407459 TI - Location and action of angiotensin II type 1 receptor in the renal microcirculation. AB - To localize angiotensin II type 1a (AT-1a) receptor and to reveal the physiological roles of angiotensin II in the renal microcirculation, we investigated the AT-1a gene deficient mice, generated by a targeted replacement of the AT-1a receptor loci by the lacZ gene (Sugaya et al, J Biol Chem 270: 18719, 1995). Immunohistochemical localization of beta-galactosidase was performed in the heterozygous mutant mice to reveal the expression sites of AT 1a. The AT-1a receptor (that is, beta-galactosidase) was expressed both in the afferent and efferent arteriolar smooth muscles and also in the mesangial cells. The effect of angiotensin II on glomerular arterioles was directly observed using the hydronephrotic mice. Angiotensin II similarly constricted both the afferent and efferent arterioles in the wild-type and heterozygous mutant mice in a dose dependent manner. This constriction was completely abolished by an AT-1 antagonist, CV-11974. In the homozygous null mutant mice, however, angiotensin II did not affect the arterioles at all. Electron microscopic studies revealed that the mesangial cells made contact with the glomerular basement membrane (GBM) at the capillary neck and also with each other in the wild-type mice. However, in the homozygous null mutant mice, the mesangial cells lost the contact either with GBM or with each other and thus the capillary neck became remarkably wider. The mesangial matrix area appeared loose and enlarged, suggesting impaired mesangial matrix formation. In conclusion, via the AT-1a receptor, angiotensin II equally constricts both the afferent and efferent arterioles and plays an essential role in maintaining the normal glomerular function and structure. PMID- 9407460 TI - Function of angiotensin II type 2 receptor in the postglomerular efferent arteriole. AB - The balance of vasucular tone between afferent (Af-Art) and efferent arterioles (Ef-Art) is a critical determinant of the glomerular hemodynamics. We recently reported that selective activation of angiotensin II type 2 receptor (AT-2R) causes dilation in the Af-Art. However, the functional role of AT-2R in the Ef Art has not been defined. In the present study, we microperfused rabbit Ef-Art in vitro, and examined the effect of angiotensin II (Ang II; 10(-11) to 10(-8) M) on the luminal diameter in the presence or absence of an AT-2R antagonist PD123319 (PD; 10(-7) M). Angiotensin II caused dose-dependent constriction in Ef-Arts, with a significant constriction occurring at 10(-11) M; at 10(-10) M the diameter decreased by 28 +/- 4% (N = 6). Pretreatment with PD significantly (P < 0.0125) augmented vasoconstrictor action of Ang II; at 10(-10) M the diameter decreased by 44 +/- 4% (N = 6). We then examined whether blockade of Ang II type 1 receptor (AT-1R) uncovers vasodilator action of Ang II mediated by AT-2R. Efferent arterioles were preconstricted by about 40% with norepinephrine and AT-1R was blocked with its selective antagonist CV11974 (CV; 10(-8) M). Angiotensin II added to preconstricted and CV-pretreated Ef-Arts caused a dose-dependent dilation; at 10(-8) M diameter increased by 28 +/- 3% (N = 5). The dilation was completely abolished by simultaneous pretreatment with PD (N = 4). Our results demonstrate that in the Ef-Art selective activation of AT-2R causes vasodilation, which opposes the vasoconstrictor action of Ang II. PMID- 9407461 TI - Nitric oxide synthase I immunoreactivity in the macula densa of the kidney is angiotensin II dependent. AB - The present study was undertaken to clarify the mechanism of the regulation of nitric oxide synthase (NOS)-I in the macula densa of the kidney. We determined the changes in NOS-I immunoreactivity of the macula densa relevant to the changes in systemic blood pressure (BP) and the renin-angiotensin system. Rats received four different types of treatment, and kidney sections were immunohistochemically stained for renin, NOS-I, and NOS-III. Plasma renin activity (PRA) and angiotensin II (Ang II) concentration were determined by radioimmunoassay. In the low-salt group, PRA, plasma Ang II, and the number of renin and NOS-I positively stained areas in the juxtaglomerular apparatus (JGA) were all increased, while BP and NOS-III in the glomerular capillaries did not change. In the desoxycorticosterone acetate (DOCA)-salt group, in contrast to the elevation of BP, PRA, plasma Ang II, and all immunohistochemical parameters were decreased. In the Ang II infusion group, BP, plasma Ang II, and the number of NOS-I positive glomeruli were increased, while PRA and renin staining were decreased. Administration of Ang II type 1 receptor (AT-1) antagonist TCV-116 significantly increased PRA, plasma Ang II, and the number of renin-positive glomeruli. However, BP, and NOS-I and NOS-III staining did not show any difference. These results clearly suggest that NOS-I in the macula densa changes in parallel with plasma Ang II, but not renin or systemic BP. PMID- 9407462 TI - Indoxyl sulfate stimulates renal synthesis of transforming growth factor-beta 1 and progression of renal failure. AB - We recently demonstrated that the administration of indoxyl sulfate (dietary protein metabolite) to 5/6-nephrectomized rats accelerated the progression of chronic renal failure by increasing the transforming growth factor (TGF)-beta 1 synthesis in the kidneys, which enhanced the renal expressions of tissue inhibitor of metalloproteinase (TIMP)-1 and type 1 collagen, leading to renal fibrosis. The aim of the present study was to clarify the mechanism by which the administration of indoxyl sulfate increases TGF-beta 1 in the kidneys of uremic rats. Since infiltrative monocytes are suggested to be an important source of TGF beta 1 in tubulointerstitial fibrosis, we examined the effect of indoxyl sulfate administration to uremic rats on the renal gene expression of intercellular adhesion molecule (ICAM)-1, which is involved in the infiltration of monocytes to kidneys. Indoxyl sulfate administration was observed to enhance the mRNA levels of ICAM-1 as well as those of TGF-beta 1, TIMP-1 and pro alpha 1 (I) collagen in the renal cortex of 5/6-nephrectomized uremic rats. In addition, we demonstrated in vitro that the addition of indoxyl sulfate significantly increased the synthesis of TGF-beta 1 in cultured proximal tubular cells. Thus, the overload of indoxyl sulfate in uremic kidneys increased the infiltration of monocytes and directly increased the synthesis of TGF-beta 1 in proximal tubular cells. PMID- 9407463 TI - Identification of genes specifically expressed in chronic and progressive glomerulosclerosis. AB - To identify genes expressed predominantly in the kidney with chronic and progressive glomerulosclerosis but not in acute and transient form of glomerulonephritis, we induced progressive glomerulosclerosis in rats by applying unilateral nephrectomy prior to injection of monoclonal anti-Thy1.1 antibody (OX 7), which cause acute and transient glomerulonephritis with single injection. In rats with nephrectomy and OX-7 injection (Nx group), proteinuria increased with time and mesangial expansion accompanied with interstitial fibrosis was recognized, whereas transient proteinuria and mesangiolysis followed by mesangial hypercellularity were seen in rats with sham operation and OX-7 injection (Sham group). Four weeks after the induction of glomerulonephritis, mRNAs were isolated from kidney cortex of both groups and used for cDNA synthesis. By subtraction hybridization of cDNAs from Nx with excess amount of those from Sham, we isolated and sequenced several genes expressed specifically in the Nx group. These included genes, which contain identical sequences with serine protease inhibitors, cytokine receptors, osteopontin as well as genes with unknown function. These genes may play important roles in the process which promotes acute glomerular damage advance to chronic and progressive glomerulosclerosis. PMID- 9407464 TI - Irbesartan lowers blood pressure and ameliorates renal injury in experimental non insulin-dependent diabetes mellitus. AB - In the present study, we investigated the effects of the angiotensin (Ang) II receptor antagonist, irbesartan, on blood pressure and renal structural injury in obese Zucker rats (OZR), an experimental model of non-insulin-dependent diabetes mellitus (NIDDM). Twenty-six-week-old OZR with established renal disease were administered either low-dose (15 mg/kg) or high-dose (50 mg/kg) irbesartan in the drinking water for a period of 18 weeks. Irbesartan caused dose-related reductions in blood pressure, and reduced by 47 to 60% the percent of glomeruli with sclerosis at 44 weeks of age (P < 0.05). In addition, irbesartan at the higher dose reduced the tubulointerstitial injury score at 44 weeks by approximately 75% (P < 0.05). By contrast, irbesartan did not significantly reduce albuminuria in OZR. The results of the present study demonstrate that the Ang II receptor antagonist irbesartan can reduce blood pressure and ameliorate glomerular and tubulointerstitial injury in an experimental model of NIDDM. PMID- 9407465 TI - Macrophage and myofibroblast proliferation in remnant kidney: role of angiotensin II. AB - Local macrophage proliferation has been shown to be a major mechanism of macrophage accumulation in several immunologically-induced animal models of renal diseases. This study has explored whether local proliferation of macrophages and myofibroblasts contribute to their accumulation in rat remnant kidney model and investigated the role of angiotensin II (Ang II) in these cellular pathological events by blocking the angiotensin II activity with ramipril, and angiotensin converting enzyme (ACE) inhibitor, or valsartan, an Ang II type 1 receptor antagonist. There was local proliferation of macrophages and myofibroblasts within renal parenchyma following renal ablation, contributing significantly to macrophage and myofibroblasts accumulation, renal dysfunction and fibrosis. Both treatment resulted in inhibition of local proliferation of macrophages and myofibroblasts and this was associated with attenuation of renal injury. In conclusion, inhibition of local macrophage and myofibroblast proliferation may be an important mechanism by which Ang II blockade attenuated renal injury following renal ablation. PMID- 9407466 TI - Candesartan prevents the progression of mesangioproliferative nephritis in rats. AB - We previously reported a new animal model of progressive glomerulonephritis induced by a single intravenous injection of the anti-Thy-1 monoclonal antibody MoAb 1-22-3 into uninephrectomized rats (Clin Exp Immunol 102: 181-185, 1995). We examined the effects of angiotensin II (Ang II) receptor antagonist (candesartan) on the clinical features and morphological lesions of this new model. By 10 weeks after induction of nephritis, untreated rats had developed hypertension, massive proteinuria, renal dysfunction, and severe glomerular injury, while uninephrectomized control rats had not. There was a significant increase in levels of glomerular protein and cortical mRNA for transforming growth factor beta (TGF-beta) and type I and type III collagens in untreated nephritic rats. Ten week treatments with candesartan and hydralazine significantly reduced blood pressure (BP) to an equal extent. Candesartan, but not hydralazine, prevented proteinuria, normalized renal function, and ameliorated glomerular injury. Candesartan also reduced levels of glomerular protein and cortical mRNA for TGF beta and type I and type III collagens, while hydralazine did not. These findings suggest that candesartan prevents progression to end-stage renal failure by modulating the effects of Ang II at least in part on the production of TGF-beta and type I and type III collagens, and not merely on systemic BP. PMID- 9407467 TI - Candesartan prevents the progression of glomerulosclerosis in genetic hypertensive rats. AB - The renin-angiotensin system (RAS) has been implicated in the development of hypertensive glomerulosclerosis. However, there are no experimental findings clearly demonstrating activation of glomerular RAS in hypertensive nephropathy. Using the stroke-prone spontaneously hypertensive rat (SHRSP) as an animal model of hypertensive glomerulosclerosis, we examined the relationship between the sequential changes in urinary albumin excretion (UAE), renal morphology, and glomerular mRNA expression for transforming growth factor-beta (TGF-beta) and fibronectin (FN) and glomerular mRNA levels for RAS components, and determined the effects of the angiotensin II (Ang II) type 1 (AT-1) receptor antagonist (candesartan) and equihypotensive hydralazine on these parameters. In SHRSP, UAE was normal at nine weeks of age and increased by 12 weeks. Plasma renin activity, plasma Ang II concentration, and angiotensin converting enzyme (ACE) activity were not higher in 9- and 12-week-old SHRSP than in WKY. RNase protection assay revealed higher glomerular mRNA levels for angiotensinogen, ACE, and AT-1a and AT 1b receptors in 9-, 12-, and 14-week-old SHRSP than in WKY. The glomerular mRNA levels for TGF-beta and FN in SHRSP were increased from nine weeks of age. SHRSP had a greater glomerulosclerosis index (GSI) at 24 weeks of age than did WKY. Administration of candesartan for two weeks, but not of hydralazine, markedly reduced UAE and normalized mRNA levels for TGF-beta, FN, and RAS components. Candesartan administration for 12 weeks virtually prevented the progression of glomerulosclerosis in rats. We conclude that in SHRSP, RAS activation and increased sensitivity to Ang II in glomeruli play important roles in the progression of glomerulosclerosis. PMID- 9407468 TI - TCV-116 inhibits interstitial fibrosis and HSP47 mRNA in rat obstructive nephropathy. AB - Unilateral ureteral obstruction (UUO) is a well established disease model leading to fibrosis of the obstructed kidney. In this model, involvement of enhanced renin-angiotensin system in the pathogenesis of interstitial fibrosis has been demonstrated. A 47-kDa heat-shock protein (HSP47) was originally identified as a collagen-binding stress protein, and is currently considered to be a collagen specific molecular chaperone that plays a pivotal role during the biosynthesis and secretion of procollagen from endoplasmic reticulum. To test if HSP47 is involved in interstitial fibrosis in UUO, we examined the expression of HSP47 mRNA in rat UUO kidneys after 12 hours. 1, 4, 7 days of obstruction. HSP47 mRNA expression was significantly increased as early as 12 hours after obstruction and was sustained at the increased level until seven days. Type I collagen mRNA significantly increased after four days of UUO. Fibrotic changes of interstitium appeared in Masson's trichrome stained section after four days. To explore the possible involvement of angiotensin II (Ang II) in HSP47 induction, the effect of Ang II receptor antagonist (TCV-116) and angiotensin converting enzyme inhibitor (lisinopril) was tested. TCV-116 or lisinopril was given to the animals orally once a day at the dose of 10 mg/kg. TCV-116 or lisinopril significantly ameliorated the fibrotic change of interstitium seven days after obstruction. HSP47 and type I collagen mRNA levels in the TCV-116- or lisinopril-treated groups were reduced to about 60% of untreated UUO. A possible involvement of HSP47 in the pathogenesis of interstitial fibrosis in UUO is suggested; however, further investigation is required to identify the signals involved in the induction of HSP47 in UUO. PMID- 9407469 TI - Long-term effects of LTB4 antagonist on lipid induced renal injury. AB - Glomerular infiltration of macrophages is a characteristic alteration of renal histopathology in hyperlipidemic renal injury. Each macrophage has two key enzymes to synthesize LTB4:5-lipoxygenase and LTA4 hydrolase. In this study we examined the long-term effects of LTB4 antagonist on real function and histopathology of spontaneously hypercholesterolemic (SHC) rat, which is model of hyperlipidemic renal injury, to see if the LTB4 antagonist could reduce the progression of renal damage. Spontaneously hypercholesterolemic rats fed a normal diet (C) developed end-stage renal failure in 26 weeks, while those fed a diet supplemented with LTB4 antagonist (E) showed normal renal function, and mild histopathological alterations (SCr: C, 1.4 +/- 0.3; E, 0.6 +/- 0.1 mg/dl, P < 0.03) without statistical differences in serum total cholesterol, body weight and blood pressure between two groups. These results suggest that LTB4 plays an important role in progression of hyperlipidemic renal injury. PMID- 9407470 TI - Pirfenidone prevents collagen accumulation in the remnant kidney in rats with partial nephrectomy. AB - Pirfenidone (PFD) is a new compound that prevents and even reverses the extracellular matrix accumulation in several organs as shown by experimental and clinical studies. In the present study, we examined the effect of PFD (500 mg/kg daily in the food) on the progression of chronic renal failure (CRF) in the 5/6 nephrectomy rat model. Proteinuria progressively increased in rats with renal ablation (C) at 12 weeks. Urinary protein excretion in PFD-treated rats (P) was numerically lower than C, but the difference did not reach statistical significance. In contrast, in the chronic phase, PFD improved renal function and reduced collagen accumulation detected by hydroxyproline content (OH-Pro) in the cortex of the remnant kidney. Although creatinine clearance decreased with time in C, the values in P were significantly better at 10 and 12 weeks. The OH-Pro in C at 12 weeks was significantly higher than that of no-ablation, sham-operated rats, whereas OH-Pro in CRF was lower in (P). Expression of mRNA for type IV and I collagen in the cortex also increased in C, but it was inhibited in (P). To study the role that TGF-beta plays in the regulatory process following CRF, we examined the expression of TGF-beta mRNA in this model. Levels of cortical TGF beta mRNA in C were significantly elevated at 12 weeks. The increase was suppressed by PFD. These results demonstrate that PFD attenuates the development of CRF by preventing collagen accumulation in this model, and suggest that PFD can be clinically useful for treating CRF. PMID- 9407471 TI - Psychodynamic psychotherapy for cancer patients. AB - Psychodynamic psychotherapy is effective as an approach to understanding the psychological conflicts and the psychiatric symptoms of cancer patients as well as to planning useful psychological interventions. The author recommends that the psychotherapist who treats cancer patients be familiar with the following: 1) the natural course and treatment of the illness, 2) a flexible approach in accord with the medical status of the patient, 3) a common sense approach to defenses, 4) a concern with quality-of-life issues, and 5) counter-transference issues as they relate to the treatment of very sick patients. Case reports illustrate the unique problems facing psychotherapists who are treating cancer patients. Further, these cases show the effective use of psychodynamic principles to inform the therapist of successful psychotherapeutic interventions. PMID- 9407472 TI - Structuring training goals for psychodynamic psychotherapy. AB - A multiaxial model that structures educational goals for psychodynamic psychotherapy has been developed. It specifies core aspects of psychodynamic psychotherapy, clusters them in categories that further define and link related areas, and presents a sequence that enables educators and students to focus on training goals in a consistent progression. This model has been used by the Director of Education as a basis for developing the curriculum, by students as a way of focusing learning and giving perspective to current work, and by supervisors to link individual teaching to the goals of the training program. This method has enhanced consistency, clarity, and efficiency in the psychotherapy program. PMID- 9407473 TI - Symptoms and character traits in patients selected for long-term psychodynamic psychotherapy. AB - In this naturalistic study of 55 outpatients selected for long-term psychodynamic psychotherapy, two Swedish assessment instruments are presented (the Karolinska Psychodynamic Profile and the Karolinska Scales of Personality), and the significance of psychodynamic criteria for the selection of patients is discussed. Thirty patients (55%) fulfilled criteria for a DSM-III-R diagnosis. The most prominent psychodynamically defined character pathology was found in the areas of coping with aggressive affects; dependency and separation; frustration tolerance; and impulse control. Some psychodynamically defined character traits, particularly poor frustration tolerance, were related to symptomatic suffering. PMID- 9407474 TI - Psychodynamic assessment and treatment of traumatized patients. AB - This article describes how psychodynamic assessment and treatment of traumatized patients can improve clinical acuity. The author describes an ego psychological, psychodynamic approach that involves 1) assessing the impact of trauma on the patient's ego defensive functioning and 2) elucidating the dynamic meaning of both the patient's presenting symptoms and the traumatic events that precipitated them. Clinical descriptions illustrate the ways in which psychodynamic psychotherapy may be particularly useful with patients whose acute symptoms develop following specific events. The author points out the advantages of an ego psychological, psychodynamic approach for her patients and the limitations of more symptom-based diagnostic assessments and treatments. PMID- 9407476 TI - The relationship between therapist-client modality similarity and psychotherapy outcome. AB - Although disparate views have been published, the theory underlying multimodal therapy suggests that therapist-client similarity would be most advantageous for treatment outcome and client satisfaction. To explore this question, 19 different therapist-client pairs were followed over 12 sessions of psychotherapy. Clients were evaluated with the Brief Symptom Inventory (BSI) after sessions 1 and 12 to determine psychotherapy outcome. Similarity was determined by computing D" statistics on therapists' and clients' responses to the Structural Profile Inventory (SPI). Similarity on the SPI predicted psychotherapy outcome, showing a statistically significant relationship with the Global Severity Index of the BSI. PMID- 9407475 TI - Using interpersonal psychotherapy (IPT) in a combined psychotherapy/medication research protocol with depressed elders. A descriptive report with case vignettes. AB - One hundred eighty subjects at least 60 years of age with recurrent unipolar major depression were recruited to participate in a depression treatment protocol. All patients received drug therapy with nortriptyline (NT) and interpersonal psychotherapy (IPT) with an experienced clinician. Acutely, 81% of subjects showed a full response to combined treatment. In the initial 127 subjects, the most common problem areas in therapy were role transition (41%), interpersonal disputes (34.5%), and grief (23%). Case vignettes are presented and discussed. The combination of IPT and NT showed a powerful antidepressant effect. IPT was readily adaptable to the needs of depressed elders. PMID- 9407477 TI - A new look at evidence about reduced cost of medical utilization following mental health treatment. 1984. PMID- 9407478 TI - Breath test diagnosis of Helicobacter pylori in peptic ulcer disease: a noninvasive primary care option. AB - BACKGROUND: Helicobacter pylori is implicated as the causative agent for most duodenal and gastric ulcers. Invasive (endoscopy and biopsy) and noninvasive (serology, breath test) methods are currently available for definitive diagnosis of infectious peptic ulcer disease. METHODS: Twenty-six patients with chronic gastritis symptoms underwent upper endoscopy, biopsy, rapid urease test, and [14C]urea breath test for the detection of H pylori. RESULTS: Twenty of 26 patients (77 percent) had biopsy-proved H pylori infection. All 20 (100 percent) with definite H pylori proved by invasive diagnosis had strongly positive results on urea breath test. Six patients with absence of H pylori on biopsy had negative urea breath test results. The urea breath test displayed 100 percent sensitivity, specificity, and predictive value compared with endoscopy and biopsy. CONCLUSIONS: [14C]Urea breath testing is comparable to endoscopy and biopsy in the diagnosis of H pylori infection and could become useful in primary care settings for noninvasive evaluation of peptic ulcer disease. PMID- 9407479 TI - Is cervicography a useful diagnostic test? A systematic overview of the literature. AB - BACKGROUND: The appropriate approach to women with mild dyskaryotic changes on Papanicolaou smear is subject to controversy. Our aim was to assess the usefulness of cervicography as a diagnostic test in detecting cervical cancer or its precursors. METHODS: We undertook an extensive literature search looking for pertinent studies of cervicography published between 1966 and 1996. Eligible studies included those in which the reference standard (colposcopy) was done on all patients. The following information was calculated: sensitivity, specificity, positive predictive value, negative predictive value, disease prevalence, and likelihood ratios. RESULTS: Cervicography has a high false-positive rate. This rate ranged from 8.2 to 61.0 percent (median 42.1 percent) for any dysplasia and 9.8 to 63.4 percent (median 50.6 percent) for high-grade cervical lesions. Likelihood ratios for a positive test result ranged from 1.0 to 10.6. Likelihood ratios for a negative result ranged from 0.02 to 1.0. CONCLUSIONS: The usefulness of cervicography is heavily dependent on the approach used to evaluate abnormal findings on a Papanicolaou smear. If a provider typically offers colposcopy to all patients with low-grade cytologic findings on a Papanicolaou smear, cervicography will decrease colposcopy use and allow for detection of cases of high-grade dysplasia missed by the index Papanicolaou smear. If a provider typically uses watchful waiting with repeat Papanicolaou smears for all patients who have low-grade cytologic findings, cervicography will substantially increase the use of colposcopy. Many of these colposcopies will be done as a result of false-positive cervigrams. PMID- 9407480 TI - Senior Team Assessment and Referral Program--STAR. AB - BACKGROUND: Although comprehensive geriatric assessment has been found to improve health and function and decrease hospital admissions, most such programs are staff-intensive and take many hours or even days. The Senior Team Assessment and Referral Program (STAR) was developed to address these two issues by using a short but comprehensive outpatient health appraisal that required only a few health professionals to complete. METHODS: Six hundred forty-nine Kaiser Permanente health plan members aged 65 years or older who received their health care at the Kaiser Permanente Medical Center, San Jose, Calif, were randomly selected during the first 12 months of the study and invited by mail to participate in STAR. Of those members contacted, 326 agreed to join the study. A nurse practitioner evaluated the health, functional, and social status of each STAR participant at an office visit once each year for the next 3 years and provided case management for those participants found to be frail or in danger of becoming frail. A control group of 764 elderly (aged 65 years and older) Kaiser members with characteristics similar to those of the STAR participants was drawn from Kaiser Permanente health plan members in San Jose. They continued to receive usual medical care throughout the study. A medical-functional profile was developed to stratify each STAR participant's overall health and functional status at each visit and case management contact. The results were entered on a grid that was used as a tracking tool throughout the study. Utilization of medical services, changes in health and function, and effects of STAR interventions on participant health behaviors were measured, and primary care physician and participant satisfaction was assessed. RESULTS: Although short-term utilization of medical services increased in the STAR group, health, function, and health behaviors improved as a result of STAR interventions. Ninety-three percent of the STAR participants described a satisfactory experience, and 71 percent were very satisfied. Sixty-five percent of primary care physicians who responded to a satisfaction questionnaire found something useful for their patients in the STAR assessment. CONCLUSIONS: STAR offers an efficient, minimally staff-intensive model for evaluating the health, functional, and social status of the 65-year-old and older age-group and intervening when they are frail or at risk of becoming frail. The improved health, function, and healthy behaviors in STAR participants and the high satisfaction rates among participants and physicians suggest that STAR would be a useful addition to the health care environment. PMID- 9407481 TI - Failed appointments in residency practices: who misses them and what providers are most affected? AB - BACKGROUND: Missed appointments can affect patient health, disrupt schedules, and result in poor utilization of resources, increased workload for staff and physicians, and lost learning opportunities for residents in training programs. METHODS: The setting was an established community-hospital-based family practice residency practice averaging 25,000 outpatient visits per year in a small northern New England town. Data from a computer-scheduling system and hospital mainframe, as well as demographic and other information contained in billing records and patient schedules, were abstracted for patients who scheduled 3962 appointments on 36 sampled days during 1995. RESULTS: The missed appointment rate during the study period was 6.7 percent. Characteristics associated with missed appointments were younger patient age (17 to 30 years), Medicaid coverage or lack of health insurance, and appointments scheduled with first-year residents or medical students. CONCLUSIONS: Attention should be given to those patients most likely to miss appointments and, in training programs, to patients seeing first year residents and medical students. It is possible that our relatively low missed appointment rate overall resulted from the nature of the practice and its environment. PMID- 9407482 TI - Practical guidelines for management of Parkinson disease. AB - BACKGROUND: This article describes a specialist's perspective on the challenge of caring for patients with Parkinson disease in a changing American health care environment that places increasing responsibility on primary care physicians. METHODS: Guidelines were developed by drawing from a combination of personal experience at a large university Parkinson disease clinic, literature review, presentations at various family practice continuing medical education conferences, and involvement as investigator in various clinical trials of drugs used in Parkinson disease treatment. RESULTS AND CONCLUSIONS: From a therapeutic standpoint, Parkinson disease can be divided into three stages--early, nonfluctuating, and fluctuating. Although the same drugs, namely, carbidopa levodopa preparations, dopamine agonists, and anticholinergic medications, are usually prescribed, their pattern of use, including frequency and dosing, varies depending on the nature of the dominant symptoms and stage of the disease. Management of Parkinson disease requires familiarity with both the disease related and drug-related components. Optimal functional efficiency for the patient is gained through striking a delicate balance between the drug regimen and the disease-related components. PMID- 9407483 TI - The incidence of Mycoplasma pneumoniae pneumonia. AB - BACKGROUND: Mycoplasma pneumoniae has been considered a pathogen for humans since the 1940s. Of the 12 species of Mycoplasma found in humans, M pneumoniae is the most widely recognized pathogen. Morbidity from M pneumoniae results from the combined direct effect of cytotoxins produced by the organisms and the indirect effect of inflammatory responses to the presence of the organisms. Several studies have reviewed the incidence of M pneumoniae pneumonia in selected populations with variable results. By using tests that were not definitive detectors of the organism, earlier studies cited have overestimated the true incidence of this infection. We reevaluate several of these early studies in the light of newer findings. METHODS: Using the key words "Mycoplasma pneumoniae," "pneumonia," "prevalence," "incidence," and "community acquired," the MEDLINE files from 1992 to the present were searched. Articles dating before 1992 were accessed from cross-reference of the more recent articles. Only clinical trials with a sample size greater than 125 were considered. RESULTS AND CONCLUSIONS: M pneumoniae pneumonia occurs in 4- to 5-year cycles and in densely populated areas. Clinical symptoms of M pneumoniae pneumonia include dry cough, sore throat, middle ear involvement, and low-grade fever, as well as additional extrapulmonary manifestations. Bullous myringitis is not a common finding in M pneumoniae infection. Diagnostic tests include cold agglutinins, complement fixation, culture, and enzyme immunoassay. A fourfold rise in M pneumoniae specific antibody in serum from acutely ill and convalescent patients remains the reference standard for diagnosing the infection. The incidence of M pneumoniae is probably lower than reported in many studies. Using tests that are not diagnostic of the infection can give a falsely elevated incidence of M pneumoniae infection in specific populations. PMID- 9407484 TI - Fourth annual Nicholas J. Pisacano Lecture. The doctor, the patient, and the home: returning to our roots. PMID- 9407485 TI - Acute myocardial infarction after use of pseudoephedrine for sinus congestion. PMID- 9407486 TI - Adie syndrome: a case report. PMID- 9407487 TI - An abbreviated model of geriatric assessment and care management: does it work? PMID- 9407488 TI - Colposcopy and referral. PMID- 9407489 TI - Withdrawal of antihypertensive medications. PMID- 9407490 TI - Progression of renal damage in human glomerulonephritides: is there sleight of hand in winning the game? PMID- 9407491 TI - Synthesis and effects of active fragments of angiotensin II. PMID- 9407492 TI - PCDAmp1, a new antigen at the interface of the embryonic collecting duct epithelium and the nephrogenic mesenchyme. AB - In the neonatal rabbit kidney nephrogenesis is not yet terminated. The ampullar collecting duct epithelium acts as an inducer that generates the nephron anlagen, however, to date the morphogenic mechanisms involved are unknown. A presupposition for successful nephron induction is the close tissue interaction between the basal aspect of the ampullar collecting duct epithelium and the surrounding mesenchyme. To gain new insights in this area we raised monoclonal antibodies (mabs), to identify specific structures localized at the tissue interface. With the generated mab CDAmp1 we found an intensive immunohistochemical reaction between the basal aspect of the ampullar collecting duct epithelium and the mesenchyme. The label was most concentrated at the ampullar tip and continuously decreased in the shaft region. In the maturing collecting duct of the neonatal kidney and in the adult renal collecting duct no immunohistochemical reaction was found. The binding pattern of mab CDAmp1 is different from that of all known collecting duct cell markers and from antibodies against known basement membrane compounds such as laminin or collagen type IV. Under in vitro conditions immunoreactivity with mab CDAmp1 was obtained using embryonic collecting duct epithelia and perfusion culture. The antigen was present in specimens treated with Iscove's modified Dulbecco's Medium (IMDM) containing 10% fetal bovine serum. Omittance of serum or hormonal treatment with aldosterone, insulin or vitamin D3 led to the disappearance of the newly detected antigen, while characteristics of the differentiated collecting duct cells were up-regulated. We conclude that the expression of PCDAmp1 is a characteristic feature of the embryonic parts of the collecting duct epithelium. It may play a pivotal role during nephron induction. PMID- 9407493 TI - Leukocyte-polytetrafluoroethylene interaction enhances proliferation of vascular smooth muscle cells via tumor necrosis factor-alpha secretion. AB - Intimal hyperplasia of vascular smooth muscle cells (VSMC) at the venous anastomosis of arteriovenous grafts represents the most common cause of vascular access failure in hemodialysis patients. Upstream release of growth factors from leukocytes activated by adhesion to the graft material may play a role in this lesion. We evaluated the effect of interaction of peripheral blood mononuclear cells (PBMC) with polytetrafluoroethylene (PTFE) on proliferation of VSMC. Vascular smooth muscle cell proliferation was significantly increased by conditioned media from human PBMC incubated with PTFE. Peripheral blood mononuclear cell adhesion to PTFE could not be antagonized by the beta 1 integrin ligand-containing peptide GRGDSP, but was attenuated by EDTA consistent with beta 2 integrin-mediated adhesion. Soluble scavenger receptor ligands at high concentrations had no effect on adhesion to PTFE excluding any contributory role of scavenger receptors in this interaction. Neutralizing antibodies to TNF-alpha significantly attenuated the mitogenic effect of PBMC/PTFE conditioned media and a marked increase in TNF-alpha secretion by PBMC on PTFE was detected by ELISA. These studies demonstrate that PBMC interaction with PTFE can promote proliferation of VSMC via increased production of TNF-alpha and perhaps other cytokines. Leukocyte interaction with PTFE causing enhanced secretion of TNF alpha and consequent VSMC proliferation may account for the development of venous intimal hyperplasia in hemodialysis patients with arteriovenous grafts. PMID- 9407494 TI - Human renal fibroblasts modulate proximal tubule cell growth and transport via the IGF-I axis. AB - To determine the paracrine interactions involved in the tubulointerstitial response to progressive renal disease, the role of insulin-like growth factor-I (IGF-I) and its binding proteins (IGFBPs) in in vitro interactions between human proximal tubule cells (PTC) and renal cortical fibroblasts (CF) were studied in primary cell culture. PTC growth and transport were increased in the presence of CF-conditioned media (CF-CM), as shown by increased thymidine incorporation, cellular protein content and sodium-hydrogen exchange (NHE) activity, to 185 +/- 31% (P < 0.01), 150 +/- 18% (P < 0.05) and 195 +/- 27% (P < 0.01) of the control values, respectively. IGF-I was produced by cultured CF at a rate of 64.6 +/- 7.5 ng/mg protein/day. Exogenous IGF-I applied to PTC provoked similar enhancement of growth and NHE activity as CF-CM and the stimulatory effect of CF-CM was blocked by specific immunoneutralization of IGF-I receptors. These receptors were threefold more abundant on PTC basolateral versus apical membranes. IGF binding proteins (IGFBP)-2 and IGFBP-3 were secreted by CF at rates of 694 +/- 88 and 1769 +/- 45 ng/mg/day, with the release of IGFBP-3 being enhanced in the presence of PTC-CM (120.0 +/- 9.7% of control, P < 0.01). Moreover, the addition of CF-CM to PTC increased cell-associated IGFBP-3 on PTC surfaces, without changes in IGF I receptor numbers or affinity and without changes in PTC mRNA for IGFBP-3. Des(1 3)IGF-I, an analog that binds to the IGF-I receptor but not to IGFBPs, provided a less potent stimulus for PTC growth compared with IGF-I, indicating that cell associated IGFBP-3 facilitates the action of IGF-I on PTC. The results support important paracrine roles for both IGF-I and IGFBPs in the interstitial regulation of proximal tubule growth and transport. PMID- 9407496 TI - Effects of integrins on proliferation and apoptosis of renal epithelial cells after acute injury. AB - We sought to determine the importance of integrins for recovery after acute tubular injury and to investigate the signal transduction pathways for integrin effects on cell cycle regulation involving proliferation and apoptosis. Primary cultures of rat renal proximal tubule epithelial cells were exposed to a superoxide-generating system to induce injury in the absence of overt necrosis. Integrin function was antagonized by the integrin recognition sequence tetrapeptide Gly-Arg-Gly-Asp (GRGD) or monoclonal antibody to beta 1-integrin. Injured cells had reduced thymidine uptake compared with normal cells. The presence of GRGD during recovery from injury caused a further 44% reduction in DNA synthesis but did not affect DNA synthesis in normal cells. Injured cells had an increased proportion of apoptosis that was further accentuated by exposed to GRGD during recovery. Integrin antagonism also stimulated apoptosis in uninjured cells. To investigate signal transduction mechanisms for this effect of integrins, inhibitors and activators of protein tyrosine kinase (PTK) and protein kinase C (PKC) were evaluated. Activation of PKC stimulated cellular proliferation, whereas inhibitors of PKC and PTK had no significant effect. Genistein, a PTK inhibitor, induced apoptosis in normal cells, mimicking the effect of integrin inhibition. On the other hand, PMA, an activator of PKC, prevented cells from becoming apoptotic when exposed to injury plus GRGD. The phosphorylation status of intracellular proteins was evaluated by immunoblotting with antiphosphotyrosine antibody. A similar pattern of decreased phosphorylation was observed after either integrin inhibition, injury, both, or PTK inhibition. These findings suggest that kinase cascades are involved in the effects of integrins on renal epithelial cell proliferation and apoptosis. After injury, an interaction between cells and the extracellular matrix is required for cells to proliferate and contribute to repair rather than to enter an apoptotic pathway. PMID- 9407495 TI - Angiotensin II modulates cell growth-related events and synthesis of matrix proteins in renal interstitial fibroblasts. AB - The renin-angiotensin system seems to play an important role in the pathogenesis of renal interstitial fibrosis. However, the potential direct effects of angiotensin II (Ang II) on cultured renal fibroblasts have been little studied. We have observed that rat renal interstitial fibroblasts (NRK 49F cell line) possess AT1 receptors coupled to intracellular calcium mobilization. Exposure of these cells to Ang II induced several short and long growth-related metabolic events mediated by the AT1 receptor, including c-fos gene expression, changes in cell cycle and cell proliferation. Activation of interstitial fibroblasts by Ang II could also contribute to extracellular matrix accumulation. Stimulation with Ang II increased mRNA expression of TGF-beta 1, fibronectin and type I collagen. In fact, Ang II enhanced fibronectin production via AT1 receptors by a process depending on autocrine TGF-beta secretion. The mechanism of some Ang II actions (calcium mobilization and fibronectin production) depended on protein kinase C and tyrosine kinase activation. We further investigated whether renal fibroblasts could express some components of the renin-angiotensin system. These cells constitutively expressed the angiotensinogen gene that was up-regulated by Ang II. Collectively, these results indicate that in renal interstitial fibroblasts Ang II causes hyperplasia and extracellular matrix production via the AT1 receptor. Ang II may initiate a positive feedback regulation of fibroblasts growth, inducing the expression of TGF-beta 1 and angiotensinogen genes. Ang II, acting directly on interstitial fibroblasts, may be implicated in the pathogenesis of renal fibrosis. PMID- 9407497 TI - Lymphocyte-derived cytokines induce sequential expression of monocyte- and T cell specific chemokines in human mesangial cells. AB - Chemokines are a family of small related proteins that play an important role in the selective recruitment of different leukocyte populations to the sites of inflammation. Human glomerular mesangial cells are potent producers of a variety of chemokines. Here we examined the kinetics of mesangial cell chemokine expression with focus on the C-C or beta chemokines monocyte chemoattractant protein-1 (MCP-1), regulated upon activation, normal T cell expressed and secreted (RANTES), macrophage inflammatory protein-1 alpha (MIP-1 alpha), and the C-X-C or alpha chemokine interleukin-8 (IL-8) in response to lymphocyte- or monocyte-derived cytokines and mesangial cell growth factors. It was found that interferon-gamma (IFN-gamma), a cytokine produced by TH1 lymphocytes, synergized with tumor necrosis factor-alpha (TNF-alpha) in RANTES expression and with IL-1 beta in MCP-1 synthesis. Time course studies revealed an early peak of mRNA expression of monocyte-specific MCP-1 upon activation with TNF-alpha in contrast to T cell-specific RANTES, which reached the highest mRNA level after 18 hours. This sequence of TNF-alpha-induced MCP-1 and RANTES expression was confirmed on the protein level. As another T-lymphocyte specific chemokine, MIP-1 alpha mRNA and protein was expressed only in response to TNF-alpha plus IFN-gamma with kinetics similar to those of RANTES expression. Finally, unlike other mesangial growth factors basic fibroblast growth factor (bFGF) induced MCP-1, RANTES, and IL-8 mRNA expression, suggesting an involvement of autocrine regulation mechanisms in mesangial chemokine expression. PMID- 9407499 TI - Ultrastructural organization of hemodialysis-associated beta 2-microglobulin amyloid fibrils. AB - Fibrils of hemodialysis-associated beta 2-microglobulin amyloid were examined by high resolution electron microscopy and immunohistochemical labeling. The amyloid containing tissues obtained through autopsy were prepared for thin section observations. In contrast to other forms of amyloid, the most conspicuous feature of these fibrils were their curved conformations. The fibril core showed ultrastructural and immunohistochemical features in common with the core of connective tissue microfibrils and of previously observed fibrils of experimental murine AA amyloidosis and familial amyloid polyneuropathy (FAP). The core was wrapped in a layer of 3 nm wide ribbon-like "double tracked" structures identified as chondroitin sulfate proteoglycan (CSPG) with immunogold labeling as well as from the results of previous in vitro experiments. Finally, the outer surface of the fibril was associated with a loose assembly of 1 nm wide filaments immunohistochemically identified as beta 2-microglobulin. This is similar to the manner in which AA protein and transthyretin filaments are associated with their respective fibrils. The results of this study provide an additional example for the concept that amyloid fibrils in general are microfibril-like structures externally associated with amyloid protein filaments. An unusual feature of the fibrils of hemodialysis-associated amyloid, however, is the presence of a peripheral layer composed of CSPG rather than of heparan sulfate proteoglycan (HSPG) as in the case of the other two amyloids above. These chondroitin sulfate chains in the outer CSPG layer may be less effective in providing rigidity to the fibril core, thus allowing for the curved conformations of beta 2-microglobulin amyloid fibrils. PMID- 9407498 TI - Reduction of insulin and triglycerides delays glomerulosclerosis in obese Zucker rats. AB - To evaluate the effect of insulin and/or triglycerides on the pathogenesis of glomerulosclerosis, acarbose (BAYg5421), an inhibitor of intestinal alpha glucosidases, was administered as a dietary admix (40 mg/100 g chow) to Zucker obese rats (ZOA), from 1.5 months until sacrifice at 1.5, 5, 8, 10 and 15 months. Obese (ZO) and lean (ZL) rats served as controls. Despite a similar food intake, ZOA weighed less than ZO at all ages. Acarbose reduced serum triglycerides at all ages, and insulin until 10 months. Glycemia remained normal in all groups. Proteinuria developed with age and to a greater degree in ZO than in ZOA rats. In ZL, a faint proteinuria appeared only in the oldest animals. Glomerulosclerosis, tubular and interstitial lesions rapidly affected ZO kidneys. These lesions were reduced in ZOA until 10 months. Acarbose did not modify the hypertrophy of the glomeruli that developed after three months, but slowed down the expansion of the mesangial domain seen in ZO. Thus, by reducing the amount of ingested glucose, acarbose yielded a normal glycemia with a lesser production of insulin and reduced renal impairment. Therefore, insulin could be a key factor involved in the pathogenesis of glomerulosclerosis, either directly or through a control of triglyceride concentrations. PMID- 9407500 TI - Subtotal nephrectomy alters tubular function: effect of phosphorus restriction. AB - Few studies have examined tubular function after subtotal nephrectomy (Nx) and conservative treatments. The effects of 70% and 80% Nx (associated with dietary phosphate restriction in the latter case) on the apical brush border membrane (BBM) enzymes 5'-nucleotidase, gamma glutamyl-transferase and alkaline phosphatase, and one BBM Na-phosphate cotransporter (NaPi-2) were studied in rats after a six week period. Changes in activity and mRNA abundance of the BBM enzymes and in NaPi-2 protein and mRNA abundance were compared with changes in the distal markers of Na,K-ATPase activity and epidermal growth factor (EGF) production. The activity, but not the mRNA of BBM enzymes, was moderately reduced by the 70% Nx. Both the mRNA and activity of gamma glutamyl-transferase and alkaline-phosphatase were decreased in the 80% Nx, and the NaPi-2 mRNA, protein and Na,K-ATPase activities were also reduced. These effects (except for 5'nucleotidase and Na,K-ATPase) were partly reversed by phosphate restriction. Overproduction of EGF occurred after the 70% Nx, was blunted in the 80% Nx, and then partially restored by phosphate restriction. Aggravation of tubular alteration was associated with enhanced renal hyperplasia (increased DNA mass), reduced GFR and hyperphosphatemia, and high PTH levels, but reduced cAMP excretion. Improvement following phosphate restriction was associated with reduced hyperplasia and lowering of phosphatemia and PTH levels. These data demonstrate that Nx selectively affected BBM function through transcriptional changes that were partially reversed by phosphate restriction. Regulatory factors involved in these changes may include intracellular phosphate content and growth factors, but not the PTH effects that are impaired in chronic renal failure. PMID- 9407501 TI - Inhibition of glomerular mesangial cell proliferation and extracellular matrix deposition by halofuginone. AB - Mesangial cell proliferation, increased deposition of collagen, and expansion of the mesangial extracellular matrix (ECM) are key features in the development of mesangioproliferative diseases. Halofuginone, a low molecular weight anti coccidial quinoazolinone derivative, inhibits collagen type alpha 1(I) gene expression and synthesis. We investigated the effect of halofuginone on both normal and SV40 transformed mesangial cell proliferation, collagen synthesis, and ECM deposition. Proliferation of both cell types was almost completely inhibited in the presence of 50 ng/ml halofuginone. The cells were arrested in the late G1 phase of the cell cycle and resumed their normal growth rate following removal of the compound from the culture medium. The antiproliferative effect of halofuginone was associated with inhibition of tyrosine phosphorylation of cellular proteins. Similar results were obtained whether the mesangial cells were seeded on regular tissue culture plastic or in close contact with a naturally produced ECM resembling their local environment in vivo. Halofuginone also inhibited synthesis and deposition of ECM by mesangial cells as indicated by a substantial reduction in 14C-glycine and Na2(35)SO4 incorporation into the ECM, and by the inhibition of collagen type I synthesis and gene expression. It is proposed that by inhibiting collagen type I synthesis and matrix deposition, halofuginone exerts a potent antiproliferative effect that may be applied to inhibit mesangial cell proliferation and matrix expansion in a variety of chronic progressive glomerular diseases. PMID- 9407503 TI - Influence of endotoxin contamination on anti-GBM antibody induced glomerular injury in rats. AB - It is accepted that the main determinant of glomerular injury in experimental nephrotoxic nephritis is the administered dose of anti-glomerular basement membrane (GBM) antibody. However, there are other factors that can enhance the severity of such injury including small doses of bacterial lipopolysaccharide (LPS). In the present study, we have assessed whether preparations of anti-GBM antibody contaminated with different concentrations of endotoxin could influence the severity of glomerular injury in the heterologous phase of nephrotoxic nephritis. We have also examined the efficacy of different laboratory methods to isolate an endotoxin-free anti-GBM antibody, and to purify anti-GBM antibody preparations from endotoxin. Preparations of anti-GBM antibody (nephrotoxic globulin) isolated from nephrotoxic serum by the sodium sulphate precipitation method contained variable concentrations of endotoxin. Administration of these preparations in equal doses into clean rats, which had no established acute phase response, markedly aggravated the severity of glomerular injury. However, preparations contained less than 50 pg/ml of endotoxin appeared to have no significant effect on such injury. Furthermore, isolation of anti-GBM antibody from nephrotoxic serum by affinity chromatography, using Staphylococcus protein-A column, proved to be a reliable method not only for the isolation of an IgG (nephrotoxic antibody) free from other serum contaminants, but also for purification of endotoxin contaminated preparations of anti-GBM antibody. These observations have practical implications in studying models of nephritis as our results show that the glomerular injury, which is usually considered to be a sole function of the mass of antibody bound to GBM, is profoundly influenced by minor endotoxin contamination of the anti-GBM antibody. PMID- 9407502 TI - Renal microvascular injury induced by antibody to glomerular endothelial cells is mediated by C5b-9. AB - We have recently developed a model of thrombotic microangiopathy with injury to the glomerular endothelial cell (GEN) induced by heterologous antibody to rat GEN. In addition to GEN injury rats developed glomerular platelet aggregation and fibrin deposition, acute renal failure, and acute tubular necrosis with interstitial inflammation. To study the role of complement in mediating this lesion, we induced the disease in normal complement PVG rats and measured the effects of generalized complement depletion with cobra venom factor (CVF) and of selective C6 deficiency using genetically C6 deficient PVG animals. Complement sufficient rats developed severe endothelial injury accompanied by platelet aggregation, fibrin deposition, decrease in endothelial cells assessed by antibody staining in the glomerulus, and macrophage infiltration. These changes were associated with marked reduction in renal function. These features were either absent or markedly diminished in complement depleted or C6 deficient rats. This demonstrates that C5b-9, the terminal product of activation of the complement cascade, plays an important role in the pathogenesis of this immune renal microvascular endothelial injury model. Thus, the complement system may play a pathogenic role in renal microvascular diseases such as thrombotic microangiopathy. PMID- 9407504 TI - Renal osmotic stress-induced cotransporter: expression in the newborn, adult and post-ischemic rat kidney. AB - The renal osmotic stress-induced cotransporter (ROSIT), a new putative member of a family of organic solute transporters, is highly expressed in the kidney. Our in situ hybridization data now reveal that large amounts of ROSIT mRNA can be found in the S3 segment of the proximal tubule. In the developing kidney, ROSIT mRNA is expressed after the S-shaped body stage. Because the S3 segment is the major site of damage in the post-ischemic kidney, we evaluated alterations in ROSIT mRNA expression after ischemic acute tubular necrosis. Renal osmotic stress induced cotransporter mRNA levels were already decreased eight hours post ischemia. At seven days post-ischemia, ROSIT mRNA reappeared in a mosaic pattern in the regenerating S3 segment, being fully expressed three weeks after the insult except for focal areas. The exact localization of this putative osmolyte transporter in the kidney, together with that of other known osmolyte transporter will contribute to a better understanding of the mechanism of medullary osmolyte accumulation and its vectorial transport. PMID- 9407506 TI - Dependence of upper limit of metastability on supersaturation in nephrolithiasis. AB - Formation of renal stones requires supersaturation (SS) high enough to induce crystallization; such a SS is referred to as the upper limit of metastability (ULM). The ULM for calcium oxalate (CaOx) or calcium phosphate can be measured by adding oxalate or calcium to urine, respectively, and noting the point at which overt crystallization occurs as evidenced by clouding. In principle, the urine should be more prone to form stone crystals as its SS approaches the ULM, and the SS ULM distance has been used as an index of stone forming potential. In addition, one would expect the ULM and initial SS to be unrelated, as the starting urine SS has no apparent link to the amount of calcium or oxalate that urine can dissolve without leading to crystal formation. However, in rats, we have found a surprising correlation between ULM and SS, such that ULM appears to rise with initial SS, for CaOx, and, to a lesser extent, for brushite (Br), a typical calcium phosphate initial phase. In this study, we measured CaOx and Br ULM, and SS, in urine of 50 patients and 11 normal people, to determine if ULM and SS were correlated, as in rats, and to explore the relationship between SS and ULM. We found the same dependence of ULM on SS as in rats, for both CaOx and Br, and found no differences between patients and normal people with respect to this dependency. However, for Br, patients showed a lower ULM than normals, but the same initial SS, meaning that patients were closer to their crystal formation threshold than normals. Treatments for stones had no apparent effect on the SS ULM dependency. We conclude that in humans, as in rats, ULM is related to initial SS, and that this relationship is the same in patients as in normals for CaOx, but shifted in a stone forming direction for Br among patients. The ULM-SS interaction is unaffected by contemporary conventional stone treatments, and is more marked for CaOx than Br. The mechanisms of the dependence are unknown. The smaller difference between ULM and initial SS for Br in patients than normal supports prior evidence suggesting a defect in stone patients that could lead to calcium phosphate crystallization, subsequent nucleation of CaOx, and stone disease. PMID- 9407505 TI - Colocalization of demethylating enzymes and NOS and functional effects of methylarginines in rat kidney. AB - NG-monomethylarginine (L-NMA) and asymmetric NG, NG-dimethylarginines (ADMA) are endogenous inhibitors of cellular L-arginine uptake and/or nitric oxide (NO) synthesis that are implicated in renal parenchymal and Dahl salt-sensitive hypertension. Since the L-arginine:(L-NMA + ADMA) ratio determines NO synthase (NOS) activity, we compared the immunohistochemical distribution of NOS with NG, NG-dimethylarginine dimethylaminohydrolase (DDAH), which inactivates dimethylarginines (DMA) and L-NMA by hydrolysis to L-citrulline. Neuronal NOS (nNOS) was expressed predominantly in tubular epithelial cells of macula densa (MD), endothelial NOS (eNOS) in vascular endothelial cells (EC), and inducible NOS (iNOS) quite widely in tubular epithelium, including proximal tubules (PT), thick ascending limbs of Henle (TAL), distal convoluted tubule and intercalated cells (IC) of the collecting duct. Immunostaining for DDAH was present in PT, TAL, MD, and IC, and was also present in the glomerulus, Bowman's capsule, and endothelium of blood vessels. DDAH was detected in small vesicles of TAL and PT by electron microscopic (EM) immunocytochemistry. To study the effects of methylarginines on tubuloglomerular feedback (TGF) response, vehicle or methylarginines (10(-3) M) were added to artificial tubular fluid (ATF) perfused orthogradely from the late PT at 40 nl. min-1 while assessing changes in glomerular capillary pressure from proximal stop flow pressure (PSF). Whereas the maximal TGF responses were unchanged by vehicle (delta TGF 0 +/- 0%) or symmetric DMA (SDMA; +1 +/- 2%, NS), they were enhanced by L-NMA (+22 +/- 4%, P < 0.001) and asymmetric DMA (ADMA; +28 +/- 3%, P < 0.001). Since L-arginine transport can regulate renal epithelial NO generation, methylarginines (10(-3) M) or vehicle were co-perfused orthogradely with [3H]-L-arginine from the late PT and collected at the early distal tubule to study arginine uptake from the perfused loop of Henle. All methylarginines reduced fractional loop [3H] absorption significantly (P < 0.001; vehicle, 84 +/- 6; ADMA, 49 +/- 6; SDMA, 56 +/- 6; L-NMA, 41 +/- 6%). In conclusion, sites of DDAH expression in the vasculature or nephron are all sites of expression of an isoform of NOS. L-NMA, ADMA, and SDMA all inhibit renal tubular L-arginine uptake, whereas L-NMA and ADMA, but not SDMA, enhance TGF responses. Therefore, DDAH may regulate the cellular L-arginine: methylarginine levels in specific renal cells, thereby governing cell-specific L-arginine uptake and NO generation in renal tubular epithelium. PMID- 9407507 TI - A quantitative analysis of sodium transport and removal during peritoneal dialysis. AB - To quantitatively evaluate peritoneal sodium transport, the diffusive mass transport coefficient (KBD) and sieving coefficient (S), as well as the mass of sodium transported by diffusion (DM), by convection (CM) and by fluid absorption (AM) and the total sodium mass removed (RM) were calculated during a series of single dwell studies in CAPD patients. A six-hour dwell study was performed in 68 patients using 2 liter of 1.36% (N = 13), 2.27% (N = 9) or 3.86% (N = 46) glucose dialysis fluid with 131I-albumin as the intraperitoneal volume marker. The patients in whom the 3.86% glucose dialysis fluid was applied were further divided into four transport groups according to a modified peritoneal equilibration test: high (H), high-average (H-A), low-average (L-A), and low (L) transport. There was no significant difference in KBD nor in S for sodium among different solutions. However, the removed sodium mass (RM) was significantly higher in the 3.86% (70.5 +/- 31.5 mmol) and 2.27% (36.0 +/- 21.0 mmol) solutions as compared to that of the 1.36% (-1.8 +/- 26 mmol) solution mainly due to increased both CM and DM. In general, CM was twice as high as DM. AM substantially decreased sodium removal. Among the different transport groups, the KBD and S values for sodium were significantly higher in the H group as compared to the other transport groups (both P < 0.05). However, RM was significantly lower in the H group mainly due to higher AM. Using a 3.86% glucose solution, the D/P for sodium was found to be significantly different (but only after 120 min of the dwell) between all the different transport groups. In conclusion, sodium removal in CAPD is strongly related to the fluid removal. The ultrafiltration induced convective transport (CM) and peritoneal absorption of sodium (AM) were of similar magnitude and were twice as high as the diffusive transport (DM) and both play an important role in the peritoneal sodium balance. A D/P for sodium using the 3.86% glucose solution, especially at the end of the dwell, can be used to discriminate between different transport categories of patients. High transport patients have a poor fluid and sodium removal that are likely to affect their clinical outcome. PMID- 9407508 TI - Bioimpedance norms for the hemodialysis population. AB - More than 3,000 hemodialysis patients were examined with single-frequency bioelectrical impedance analysis (BIA). Distributions of resistance, reactance, phase angle (PA), and estimates of total body water (TBW) and body cell mass (BCM) by BIA were determined, and compared with traditional laboratory markers of nutritional status. Bioimpedance parameters and body composition estimates differed significantly by age, sex, race, and diabetic status. PA and BCM correlated directly with serum creatinine, albumin, and prealbumin concentrations. Population-based norms for bioimpedance parameters and estimates of body composition are provided. PMID- 9407509 TI - Low dose angiotensin converting enzyme inhibition and glomerular permselectivity in renal transplant recipients. AB - In this study, we determined the fractional clearance of neutral polydisperse dextrans (theta D) and monodisperse dextran sulfate (theta DS) to describe glomerular size and charge selectivity in 25 renal transplant recipients with proteinuria. Thirteen were treated with low dose lisinopril for six months (group 1) and 12 patients without ACE inhibitor therapy formed group 2. Mean arterial blood pressure was stable (group 1, 112 +/- 4; group 2, 109 +/- 2 mm Hg at baseline and after 6 months) whereas creatinine clearance, glomerular filtration rate and renal plasma flow decreased nonsignificantly but were comparable at any time. Lisinopril treatment lowered filtration fraction (22 +/- 2 vs. 19 +/- 2%, P = 0.07) whereas no change was seen in group 2 (20 +/- 2%). The fractional protein excretion (mg urinary protein per day/ml creatinine clearance per day) was stable in group 1, but significantly increased in group 2. The same pattern was found for theta D larger than 56 A. theta DS was stable and consistently elevated in both groups at any time. We conclude that low dose ACE inhibitor treatment in proteinuric renal transplant recipients stabilizes glomerular size selectivity independently of its systemic hemodynamic effects. PMID- 9407510 TI - Necrosis and apoptosis of polymorphonuclear cells exposed to peritoneal dialysis fluids in vitro. AB - Conventional peritoneal dialysis (PD) fluids are known to inhibit polymorphonuclear cells (PMN) phagocytosis, oxidative burst and enzyme release. However, the relative contributions of apoptosis and/or necrosis to this dysfunction have not been examined. We investigated the effects of osmolality, glucose concentration and heat-sterilization of PD fluids on necrosis and apoptosis of PMN. Polymorphonuclear cells were isolated from 8 healthy volunteers and exposed to different PD fluids for four hours. PMN were then double-stained with Hoechst 33342 and propidium iodide to study the proportion of viable, apoptotic and necrotic cells. Transmission electron microscopy (TEM) was performed to confirm the results obtained with flow cytometry. The fluids studied were conventionally heat-sterilized 1.5% Dianeal (1.5% D), conventionally heat sterilized 4.25% Dianeal (4.25% D), 1.5% D in which the osmolality was increased to that of 4.25% D by adding mannitol (1.5% D + M), a filter-sterilized version of 4.25% D (4.25% D-F) and a 1.1% amino acid PD fluid (AA) (Nutrineal PD4). All PD fluids had their pH equilibrated (pH = 7.4) by the addition of sodium bicarbonate. Compared to PMN exposed to culture medium, a significantly higher proportion of necrosis was observed in PMN exposed to 1.5% D (P = 0.04). The 4.25% D induced greater necrosis than 1.5% D (P = 0.001), and the 4.25% D also induced significantly more necrosis (P = 0.002) compared to 4.25% D-F. These data suggest that the consequences of heat-sterilization, rather than high glucose concentration are responsible for the necrosis observed. Indeed, the proportion of necrotic PMN with 4.25% D-F was not significantly different from 1.5% D. The 1.5% D + M and AA induced significantly more apoptosis compared to 1.5% D (P = 0.006 and P < 0.05, respectively), suggesting that apoptosis can be induced by the high osmolality of PD fluids. However, 1.5% D +/- M also induced significantly more apoptosis (P = 0.007) compared to 4.25% D-F. This suggests that the apoptosis effect is specific for the osmolyte present in PD fluids, and that mannitol and amino acids induce more apoptosis than glucose. In summary, the different non-physiological components of conventional PD fluids evaluated in this study had a differential effect on PMN survival. Heat sterilization of high glucose-containing PD fluids was associated predominantly with necrosis of PMN, and high osmolality with apoptosis. PMID- 9407511 TI - Lung density for assessment of hydration status in hemodialysis patients using the computed tomographic densitometry technique. AB - The density of the lung reflects the total mass of fluid, air, and dry lung tissue per unit volume of the lung. Lung density can be measured by evaluation of attenuation of an electron beam with computed tomography (CT). This technique has been shown to be sufficiently reliable and sensitive to distinguish normal from abnormal lung water. The aim of this study was to find out whether lung density properly reflects the hydration status in hemodialysis patients in comparison with other standard methods. Fourteen hemodialysis patients, with an ultrafiltration ranging from 0.3 to 4.5 liters per session, underwent CT measurements of lung density, ultrasonographic measurements of the diameter of the inferior vena cava after quiet expiration (IVCe) and quiet inspiration (IVCi), and measurements of the hematocrit and plasma levels of the biochemical hydration markers cyclic guanosine monophosphate (cGMP) and atrial natriuretic peptide (ANP). These measurements were performed before and 3.5 to 4 hours after termination of dialysis. Quantitative estimates of lung density were obtained within pixels with CT numbers ranging between -1000 and -100 Hounsfield Units (HU), and compared with normal data from 18 normal controls. In normal controls, the lung density ranged from -800 to -730 HU. In hemodialysis patients, lung density was significantly higher than normal before dialysis (-678 +/- 96 HU, P < 0.01) and significantly decreased after dialysis (-706 +/- 92 HU, P < 0.05), indicating a decrease in fluid content of the lung. The density was normalized in 5 patients. A significant correlation was found between lung density and IVCe both before and after dialysis (r = 0.8, P < 0.01 for both). Change in density was significantly correlated to amount of ultrafiltration (r = 0.67, P < 0.01) and percent change in blood volume (r = 0.63, P < 0.05), indicating that lung density is greatly affected by changes in the extracellular fluid volume, mainly the intravascular volume. In conclusion, lung water reflects the hydration status in hemodialysis patients and can be monitored by measuring the lung density by CT. Accordingly, normalization of lung density can help to achieve a proper dry weight in these patients. PMID- 9407512 TI - Reduction of plasma apolipoprotein-B by effective removal of circulating glycation derivatives in uremia. AB - Patients with diabetes and renal insufficiency (Db/ESRD), a group subject to accelerated atherosclerosis exhibit marked increases in the levels of circulating, glycation-derived reactive substances, termed advanced glycation endoproducts (AGEs). These products have been previously shown to react covalently with apoliprotein B (ApoB) to form AGE-ApoB, a modification that results in delayed low density lipoprotein (LDL) clearance and possibly to dyslipidemia. Because the effect of hemodialysis on AGE removal was shown to be unsatisfactory, based on single intradialytic studies, we examined the effect of long-term hemodialysis therapy on serum AGE-ApoB levels, as well as on total serum ApoB of 25 Db/ESRD patients treated by two types of hemodialysis filters, the Fresenius Inc. F8, as the low flux (LF), or high-flux polysulfone AN69 (HF) for two months using an AGE-specific ELISA. At the end of eight weeks, circulating AGE-ApoB levels were reduced significantly (by 35%) from baseline (P = 0.039) in patients treated by HF compared to a modest 16% reduction noted in patients treated by LF (P = 0.05) N = 12, P = 0.047). Of note, total plasma ApoB was reduced by 27% from baseline (P = 0.02) in patients treated by HF compared to a 6% reduction noted in those treated with LF (P = 0.8). In vitro comparison of AGE mass balance, and mass adsorption by the different filters revealed that the higher efficiency of HF filter was due to greater adsorption. The association of reduced AGE-ApoB levels with a decrease in total circulating ApoB by HF and not by LF dialysis suggests: (1) a causal link between AGE clearance and dyslipidemia in diabetic ESRD, and, (2) that more efficient modes of renal replacement treatment and AGE removal could significantly benefit clinical outcome. PMID- 9407513 TI - Adrenocorticotrophic hormone lowers serum Lp(a) and LDL cholesterol concentrations in hemodialysis patients. AB - Previously, we have shown that short-term administration of adrenocorticotrophic hormone (ACTH) results in reduced concentrations of apolipoprotein B-containing lipoproteins, including lipoprotein(a), and reduced activities of hepatic lipase. These effects were observed in steroid-treated patients suffering from iatrogenic ACTH deficiency and in healthy individuals. The direct nature of the influence of ACTH on hepatic lipoprotein metabolism was confirmed by in vitro experiments. The aim of the present investigation was to study the effects of ACTH treatment on uremic patients, who exhibit disturbed lipoprotein pattern due to the slow removal of triglyceride-rich lipoproteins and who probably are ACTH resistant. Eight patients on chronic hemodialysis were studied. After one intramuscular injection of Synacthen Depot (a synthetic ACTH1-24 preparation from Ciba Geigy AG, Basel, Switzerland) 1 mg, the only change noted was a significant reduction of 26% in median lipoprotein(a) concentration. After five injections, a further decrease (65%) was found in the lipoprotein(a) concentration. Also, reductions in median concentrations of total cholesterol, low density lipoprotein cholesterol and apolipoprotein B were observed. The magnitude of these changes was 15 to 30%. In contrast to previously studied groups, no changes were observed regarding triglyceride metabolism. Significantly increased median concentration of apolipoprotein CIII was found. However, the excess apolipoprotein CIII was confined to the fraction that was not associated with apolipoprotein B. Thus, administration of ACTH to uremic patients improved their atherogenic lipoprotein profile, a fact that may have future therapeutic implications. In comparison to previously studied groups, the uremic patients responded rather slowly and not at all regarding triglyceride metabolism. PMID- 9407514 TI - Predictive measures of vascular access thrombosis: a prospective study. AB - Malfunction of permanent vascular accesses remains a cause of frequent and costly morbidity in patients receiving chronic hemodialysis (CHD). Several recommendations for routine monitoring of these permanent vascular accesses for incipient failure have been proposed. In this study, multiple indicators of incipient vascular access dysfunction, including "venous" and "arterial" pressures at serial blood flows (200 ml/min, 300 ml/min, and 400 ml/min), percent urea recirculation, Doppler ultrasound, and access blood flow by ultrasound dilution technique were simultaneously evaluated in a total of 220 vascular accesses in 170 chronic hemodialysis patients in two separate study periods (6 months apart). The rate of thrombosis was determined within the subsequent 12 weeks of each study period to assess the short-term predictive power of access thrombosis. During the period of follow-up, there were 34 thrombotic events in 172 polytetrafluoroethylene (PTFE) grafts and only one thrombotic event in 48 arterio-venous fistulas (AVF). Therefore, the statistical analysis was limited to the PTFE grafts. When grafts with thromboses were compared to those without thrombosis by univariate analysis, access blood flow measured either by ultrasound dilution technique (875 +/- 426 ml/min with thrombosis vs. 1193 +/- 677 ml/min without thrombosis, P = 0.001) or by Doppler ultrasound (762 +/- 420 ml/min with thrombosis vs. 1171 +/- 657 ml/min without thrombosis, P = 0.001) were significantly different in the two groups. There was good correlation (r = 0.79, P = 0.0001) between the blood flows determined by both techniques. The grade of stenosis determined by ultrasound was also a statistically significant predictor (P = 0.02). "Venous" and "arterial" pressures were numerically similar and were not statistically different between the accesses that did and those that did not thrombose. When multivariate analysis was used, there was a significantly increased risk of thrombosis only with decreasing access blood flow determined by ultrasound dilution techniques after adjusting for other confounding variables. When the average blood flow of all grafts (1134 ml/min) is considered as the reference access blood flow (relative risk of 1.0), the relative risk of a PTFE thrombotic event within the subsequent 12 weeks was 1.23 at a blood flow 950 ml/min, 1.67 at a blood flow of 650 ml/min and to 2.39 at a blood flow of 300 ml/min. In summary, access blood flow measured by either Dilution or Doppler is a reliable indicator of subsequent short-term thrombosis risk. Other proposed methods of evaluating access dysfunction were not useful in our patients. If simple to use, cost-effective devices to measure dialysis access blood flow become readily available, the measurement of access blood flow will likely become the method of choice for screening of PTFE vascular access dysfunction in hemodialysis patients. PMID- 9407515 TI - Dialytic nutrition: provision of amino acids in dialysate during hemodialysis. AB - Maintenance hemodialysis (MHD) patients are frequently malnourished, a condition associated with high morbidity and mortality. Amino acid (AA) losses in dialysate may enhance protein malnutrition in patients with low food intake. We studied the possibility of providing AA in dialysate during MHD to either prevent AA losses or as a nutritional supplement. Six clinically stable men were studied during three hemodialysis treatments. The first treatment was performed using the usual dialysate (0XAA). The two other treatments were performed using a dialysate containing an amount of AA equal to normal plasma AA concentrations (1XAA) or to three times the normal plasma AA concentrations (3XAA). During the OXAA treatment, the total AA losses were 10.0 +/- 0.9 (SEM) grams (g) and the total AA concentrations in plasma decreased by 49 +/- 4%. During the 1XAA treatment, the total AA balance was +0.8 +/- 1.8 g and there was no significant change in the postdialysis plasma total AA. With the 3XAA treatment, the patients gained 36.9 +/- 4.1 g of AA during the hemodialysis treatment and the plasma total AA levels increased by 45 +/- 9%. No side effects were observed. These findings indicate that it may be feasible to provide AA supplements to MHD patients by adding AA to hemodialysate. PMID- 9407516 TI - Dipyridamole inhibits PDGF- and bFGF-induced vascular smooth muscle cell proliferation. AB - Dipyridamole is the only pharmacologic agent demonstrated to reduce polytetrafluoroethylene (PTFE) graft occlusion in hemodialysis patients. However, the mechanism of action of dipyridamole in preventing graft occlusion is unknown. The purpose of this study was to examine the direct effects of dipyridamole on both platelet-derived growth factor (PDGF) and basic fibroblast growth factor (bFGF)-induced vascular smooth muscle cell (VSMC) proliferation. Human aortic smooth muscle cells were grown to confluence in 96 well plates. A total of 5 x 10(-6) molar dipyridamole, PDGF 10 ng/ml, or bFGF 10 ng/ml were added to appropriate wells at the start of each experiment. Cell proliferation at 48 hours was determined using tritiated thymidine uptake. Intracellular cyclic AMP (cAMP) was measured using a competitive enzyme immunoassay. Treatment of VSMC with 5 microM dipyridamole dramatically reduced basal proliferation rates compared to controls [5229 +/- 1131 counts per minute (CPM) versus 387 +/- 68 CPM, P < 0.001]. Treatment with dipyridamole also reduced PDGF-stimulated VSMC proliferation (7311 +/- 1655 CPM vs. 593 +/- 110 CPM, P < 0.001) as well as the response to bFGF (5632 +/- 1270 CPM vs. 310 +/- 31 CPM, P < 0.001). Treatment of VSMC with either 5 or 20 microM dipyridamole did not change intracellular cAMP levels. Furthermore, the addition of dibutyryl cAMP to VSMC demonstrated only a modest inhibitory effect on proliferation. We conclude that dipyridamole inhibits both PDGF- and bFGF-stimulated VSMC proliferation. The effects of dipyridamole on VSMC proliferation do not appear to be entirely mediated by changes in intracellular cAMP concentrations. The direct effect of dipyridamole on VSMC proliferation may account for its efficacy in reducing PTFE graft thrombosis in hemodialysis patients. PMID- 9407518 TI - Increased plasma concentrations of LDL-unbound apo(a) in patients with end-stage renal disease. AB - Lipoprotein(a) [Lp(a)] and its characteristic glycoprotein apolipoprotein(a) [apo(a)] are risk factors for atherosclerosis in the general population. Patients with renal disease show an elevation of Lp(a). Recent studies have described an arteriovenous difference of Lp(a) in the renovascular bed as well as the plasma derived fragmented LDL-unbound apo(a) in urine, suggesting that the kidney is involved in the metabolism of Lp(a). We therefore investigated whether patients with chronic renal failure have higher levels of LDL-unbound apo(a) and whether this could account for the increased Lp(a) concentrations in these patients. In addition, we studied the possible generation of apo(a) fragments in vitro by mimicking uremic plasma conditions and by investigating the assembly of Lp(a) in cell culture experiments. Patients treated by hemodialysis (N = 185) and by continuous ambulatory peritoneal dialysis (CAPD; N = 20) had markedly elevated absolute (1.22 +/- 1.55 mg/dl and 2.14 +/- 2.86 mg/dl) as well as relative (7.5% and 7.3%) amounts of LDL-unbound apo(a) in comparison to controls (0.46 +/- 0.48 mg/dl or 4.5%). Following renal transplantation the absolute amount decreased significantly. Lp(a) plasma concentration was the most important determining variable for the absolute amount of LDL-unbound apo(a) and showed a positive correlation in both hemodialysis patients (r = 0.85) and controls (r = 0.92). In vitro experiments demonstrated that "uremization" of plasma samples did not generate a higher amount of LDL-unbound apo(a). Although LDL of renal patients has different chemical and structural properties as compared to control LDL, the extracellular assembly of Lp(a) did not differ between patients and controls. Therefore, the higher amounts of LDL-unbound apo(a) found in renal disease are not caused by an impaired assembly of Lp(a), but rather indicate a catabolic role of the kidney for LDL-unbound apo(a) as was already shown for Lp(a). Despite a small contribution, these elevated levels cannot explain the higher Lp(a) values found in patients with end-stage renal disease. PMID- 9407517 TI - CD28-b7 blockade in organ dysfunction secondary to cold ischemia/reperfusion injury. AB - Ischemic injury to cadaver organs is a major risk factor for development of chronic organ dysfunction. We have recently shown that the B7 costimulatory pathway plays a critical role in early organ dysfunction developing after renal cold ischemia/reperfusion injury. We extended these observations to investigate the role of this pathway in the development and progression of chronic organ dysfunction following such injury. Uninephrectomized rats which underwent cold ischemia/reperfusion injury developed progressive proteinuria as compared to uninephrectomized controls. Animals treated with CTLA4Ig, which blocks B7 costimulation, starting on the day of injury had significantly better long-term survival and developed significantly less proteinuria than control animals treated with control Ig. RT-PCR analysis of kidney tissue showed significant reduction in expression of activation and inflammatory cytokines, chemoattractants, and growth factors, as compared to controls. Delaying administration of CTLA4Ig for one week, but not four weeks, after injury was still effective in ameliorating development of progressive proteinuria. Interestingly, selective blockade of B7-1 by a mutant form of CTLA4Ig had no effect on early or chronic organ dysfunction. These findings indicate the long term functional and molecular consequences of experimental cold ischemia/reperfusion injury, and suggest that B7-2 is critical in the development of organ dysfunction following ischemic injury, even in the absence of alloantigen. PMID- 9407519 TI - Drug-targeting to the kidney: renal delivery and degradation of a naproxen lysozyme conjugate in vivo. AB - A renal-specific controlled release of an active drug may enable a reduction of the required dose and may provide a reduction of extra-renal toxicity. To achieve renal specific targeting of the NSAID naproxen, the low-molecular-weight protein (LMWP) lysozyme was employed as carrier since it is mainly taken up and catabolized in the proximal tubules of the kidney. A conjugate was synthesized with an average coupling degree of 2 mol naproxen per 1 mol lysozyme in which the drug was directly coupled to the protein via a peptide bond. First, we investigated whether naproxen conjugation affects the renal disposition of lysozyme. As native lysozyme, the conjugate was predominantly and rapidly (within 20 min) taken up by the kidney. The subsequent decrease in renal content reflecting the renal degradation of the conjugated lysozyme molecules appeared also to be similar to that of native lysozyme with a half life of four hours. Second, the effect of lysozyme conjugation on the body distribution of naproxen was studied. An important observation with regard to the aimed reduction in extra renal side effects was that no detectable amounts of free naproxen were present in the plasma after administration of conjugate. Conjugation of naproxen to lysozyme resulted in a pronounced (70-fold) increase of naproxen accumulation in the kidney. In agreement with the protein disposition study, the conjugate was rapidly taken up by the kidney and subsequently degraded. In conclusion, renal selective targeting of the NSAID naproxen can be obtained by conjugation with the LMWP lysozyme. This concept of drug delivery to the kidney has the potential to improve drug efficacy and safety. PMID- 9407520 TI - Serum oxalate microassay using chemiluminescence detection. AB - Determination of serum oxalate concentration is important for the diagnosis and monitoring of hyperoxalurias, and extends to patients with all types of renal disease. Approximately 5 to 10 ml of blood is required for each test by conventional methods, and the test is not adapted for use in children. We developed a highly sensitive method that limits the volume of blood required for the test. This new and sensitive tool to detect H2O2 can be successfully substituted for the conventional, and expensive, colorimetric reaction to accurately analyze oxalate concentration. PMID- 9407521 TI - Hemodialysis vascular access: an ounce of prevention. PMID- 9407522 TI - A role for T cell co-stimulation in ischemic acute renal failure? PMID- 9407523 TI - Epidemic hemolytic-uremic syndrome in children. PMID- 9407524 TI - DNase I footprinting. PMID- 9407525 TI - Quantitative DNA footprinting. PMID- 9407526 TI - Uranyl photoprobing of DNA structures and drug-DNA complexes. PMID- 9407527 TI - Diethylpyrocarbonate and osmium tetroxide as probes for drug-induced changes in DNA conformation in vitro. PMID- 9407528 TI - Footprinting studies with nucleosome-bound DNA. PMID- 9407529 TI - A gel mobility shift assay for probing the effect of drug-DNA adducts on DNA binding proteins. PMID- 9407530 TI - An oligonucleotide crosslinking assay for the analysis of individual drug-binding sites. PMID- 9407532 TI - Transcriptional footprinting of drug-DNA interactions. PMID- 9407531 TI - DNA polymerase inhibition assay (PIA) for the detection of drug-DNA interactions. PMID- 9407533 TI - Determination of the DNA sequence specificity of alkylation damage using cleavage based assays. PMID- 9407534 TI - PCR-based methods for detecting DNA damage and its repair at the subgene and single nucleotide levels in cells. PMID- 9407535 TI - Analysis of DNA-binding antibodies. PMID- 9407536 TI - Immunofluorescent staining of chromosomes with DNA-binding antibodies. PMID- 9407537 TI - Optical absorbance and fluorescence techniques for measuring DNA-drug interactions. PMID- 9407538 TI - Evaluation of drug-nucleic acid interactions by thermal melting curves. PMID- 9407539 TI - Electric dichroism. PMID- 9407540 TI - Calorimetric techniques for studying drug-DNA interactions. PMID- 9407541 TI - Methods for the studies of drug dissociation from DNA. PMID- 9407542 TI - Liver development in the rat and in man during the embryonic period (Carnegie stages 11-23). AB - Hepatic structures appearing during embryonic Carnegie stages 11-23 were analyzed and compared in OFA-IOPS rat and human embryos. The group of rats--crown-rump length (CRL) 2-16 mm, 10-16 days postcoitus--was composed of 127 specimens (52 of stages 11-12, 55 of stages 13-19, and 20 of stages 20-23), the human group of 9 embryos at stages 14-23--CRL 5-31 mm, age 32-57 days--and human stages 11-13 were described according to former literature. The specimens were subjected to serial histological sections with graphic reconstructions. In both series, stage 11 was characterized by hepatic diverticulum development, stage 12 and thereafter by cellular differentiation (septum transversum giving the liver stroma and hepatic diverticulum the hepatic trabeculae), and stage 13 by epithelial cord proliferation enmeshing stromal capillaries. From stage 14, the hepatic gland and its vascular channels presented considerable enlargement while hematopoietic function appeared. From this stage, the development of cystic primordium, never present in rat, was constant in man. At stage 18, after a period of obturation due to epithelial proliferation, the bile ducts became reorganized and ensured the continuity between liver cells and gut. From stages 18 to 23, biliary ductules developed in periportal connective tissue producing ductal plates that received biliary capillaries. Except for gallbladder, similarity and presence of the same hepatic structures in man and rat during the embryonic period stages 11 23 permit us to consider the rat as a good experimental model for liver development, for example, in studies on teratology and congenital anomalies. PMID- 9407543 TI - Development and differentiation of bile ducts in the mammalian liver. AB - The development and differentiation of bile ducts in the human and rodent liver are reviewed. The liver primordium develops as a ventral diverticulum in the anterior intestinal portal region, which consists of endodermal and mesodermal components. The endodermal cells differentiate into hepatocytes and all epithelial cells of the bile ducts in the adult liver. The gallbladder and extrahepatic bile ducts also start to develop from hepatic endodermal cells and hepatoblasts just after liver primordium formation. The gallbladder and cystic duct do not develop through hepatic development in the rat. Intrahepatic bile ducts are formed from periportal hepatoblasts forming the "ductal plate" and expressing alpha-fetoprotein, and albumin and bile duct-specific cytokeratin and develop independently of extrahepatic bile duct formation. The first sign of intrahepatic bile duct differentiation is the increased expression of bile duct specific cytokeratin and large lumina formation in periportal hepatoblasts, and then deposition of basal laminar components occurs on the basal side. Their development takes place discontinuously along portal veins at the early stage of development, and they then become confluent through development. Periportal connective tissue, glucocorticoid hormones, and basal laminar components may play important roles in the differentiation of bile ducts. PMID- 9407544 TI - Development of hepatic sinusoidal structure with special reference to the Ito cells. AB - To elucidate sinusoidal cell structure and function under normal conditions and their behavior in diseased settings, an understanding of their developmental aspects is needed. At day 10 of gestation in mice and rats or at 5 weeks of gestation in humans, the hepatic cords grow into the mesenchymal tissue of the septum transversum, and the primitive sinusoidlike structure is simultaneously observed between the liver cell cords. In the margin of the growing liver primordium, mesenchymal cells in the septum transversum are trapped in the subendothelial space. These subendothelial cells are at the early stages of organogenesis and become progenitors of the Ito cells. By days 12-14 of gestation in mice and rats or 8 weeks of gestation in humans, the basic structure of the sinusoids has developed. Embryonic hepatic sinusoids are usually lined by a continuous endothelium without basement membranes, and an incompletely fenestrated sinusoid appears at the middle gestational stage. In the late gestational stages, the Ito cells exhibit myofibroblastlike features in humans, mice, and rats. In association with this event, perisinusoidal reticular networks are gradually intensified. After birth until days 4-5 in mice and rats, the sinusoidal and perisinusoidal structures are almost completely formed, although slight morphological differences from those in adult livers still exist. What happens to sinusoidal endothelial cells and Ito cells in hepatic fibrosis cirrhosis of the adult may be a deviated or uncontrolled occurrence of what goes on during the fetal period, i.e., a continuous nonfenestrated sinusoidal lining in the early embryonic stage and a myofibroblastlike transformation of Ito cells in late fetal life. PMID- 9407545 TI - Development, differentiation, and maturation of Kupffer cells. AB - Primitive macrophages first develop in the murine and human yolk sac and then differentiate into fetal macrophages. Primitive or fetal macrophages enter the blood stream and migrate into the fetal liver. Fetal macrophages possess a high proliferative capacity and express antigens and peroxidase activity of resident macrophages with the progress of gestation; they become mature and then transform into Kupffer cells. In contrast, myelopoiesis and monocytopoiesis are not active in yolk sac hematopoiesis and in the early stages of hepatic hematopoiesis. Precursor cells of primitive or fetal macrophages exist and granulocyte/macrophage colony-forming cells develop in the yolk sac and in the early stages of fetal liver development, whereas macrophage colony-forming cells emerge and increase later in fetal liver development. In vitro, similar colonies were formed from each fetal hematopoietic cell in the presence of different macrophage growth factors. During culturing of the yolk sac cells and hepatic hematopoietic cells on a monolayer of mouse stromal cell line, ST2, primitive or fetal macrophage colonies developed before the formation of monocyte colonies, suggesting the existence of a direct pathway of differentiation from primitive macrophages into fetal macrophages during ontogeny. In severely monocytopenic mice induced by the administration of strontium-89, Kupffer cells have a proliferative capacity and are maintained by self-renewal. In macrophage colony stimulating factor (M-CSF)-deficient (op/op) mice, the number of Kupffer cells is reduced, and they are characterized by immature morphology and a proliferative potential similar to that of primitive or fetal macrophages during ontogeny. Immediately after the administration of M-CSF to op/op mice, Kupffer cells start proliferating and become mature. This finding indicates that M-CSF plays an important role in the differentiation and proliferation of Kupffer cells. PMID- 9407546 TI - Development of neuropeptide Y innervation in the liver. AB - Hepatic neuropeptide Y (NPY) innervation was studied by immunohistochemistry in various mature vertebrates including the eel, carp, bullfrog, turtle, chicken, mouse, rat, guinea pig, dog, monkey, and human. In addition, an ontogenetic study on hepatic NPY was made in developing mice and guinea pigs. In all species examined except the eel, NPY-like immunoreactivity was detected in nerve fibers. In the carp, bullfrog, turtle, chicken, mouse, and rat, NPY-positive fibers were distributed around the wall of hepatic vessels and the bile duct of the Glisson's sheath. The density of NPY-positive fibers increased with evolution. However, in the guinea pig, dog, monkey, and human, numerous NPY-positive fibers were observed not only in the Glisson's sheath but also in the liver parenchyma. Positive fibers formed a dense network that surrounded the hepatocytes. The present immunoelectron microscopic study has confirmed that NPY-positive terminals are closely apposed to hepatocytes. Ontogenically, NPY-positive fibers were first found in the embryonic liver of 19-day-old mice. Positive fibers increased with age, and the highest peak was seen 1 week after birth. However, NPY-positive nerve fibers were present abundantly in Glisson's sheath and in the hepatic parenchyma of neonatal (3 and 7 days old) guinea pigs in a distribution similar to that in mature animals. This ontogenetic pattern suggests that NPY plays a certain role in the developing liver. PMID- 9407547 TI - Expression and potential role of the extracellular matrix in hepatic ontogenesis: a review. AB - Studies from a number of laboratories have provided information on the temporal and spatial expression of a variety of extracellular matrix (ECM) components in the developing liver and insight into their potential roles in hepatogenesis. Collagen type IV and laminin are present in the basement membranes of the capsular mesothelium, vascular structures of the portal and hepatic vein branches, and the ductular elements of the developing liver. The mesothelial, vascular, and ductular epithelial cells synthesize laminin and type IV collagen. In contrast, fibronectin and type I collagen are restricted to the adjacent or surrounding interstitium of those ductal and vascular elements, but are not within the basement membrane proper. The hepatic perisinusoidal space (Space of Disse) of the fetal rat develops a delicate extracellular matrix by 12.5 days of gestation, which is characterized by banded collagen fibrils and bundles associated with filamentous and flocculent material. Fibronectin, laminin, and collagen types I, III, and IV are present in the developing perisinusoidal space by this early gestational date, with laminin being the most prevalent component detected. The laminin chains localized to that region in the fetal/neonatal period are alpha 2, beta 1, beta 2, and gamma 1, whereas the alpha 1 chain of laminin is absent from the developing Space of Disse. Similar data have been reported on the laminin phenotype in the perisinusoidal space during hepatic regeneration. Electron microscopy immunohistochemistry studies have demonstrated that the sinusoidal lining cells and hepatocytes synthesize these ECM proteins during hepatogenesis. By 6 to 8 weeks of postnatal life, laminin is not detectable in the perisinusoidal space. Both the transient expression of laminin and the similarity of the laminin chain phenotype expressed in the perisinusoidal space in the developing and regenerating liver suggests a role for this protein in the organization of the hepatic lobule in those forms of hepatic morphogenesis. PMID- 9407549 TI - Detection and differentiation of the six Brucella species by polymerase chain reaction. AB - BACKGROUND: Brucelosis is a severe acute febrile disease caused by bacteria of the genus Brucella. Its current diagnosis is based on clinical observations that may be complemented by serology and microbiological culture tests; however, the former is limited in sensitivity and specificity, the latter is time consuming. To improve brucelosis diagnosis we developed a test which is specific and sensitive and is capable of differentiating the six species of Brucella. MATERIALS AND METHODS: Four primers were designed from B. abortus sequences at the well-conserved Omp2 locus that are able to amplify the DNAs of all six species of Brucella. RESULTS: Our test detected all six species of Brucella. Their differentiation resulted directly from differences in the amplification patterns or was achieved indirectly using a RFLP present in one of the PCR products. The sensitivity and specificity of the new test were then determined; it was applied successfully in confirming the diagnosis of a patient whose clinical history and serology indicated infection with Brucella. CONCLUSIONS: The results make possible the use of a PCR test for Brucella detection and differentiation without relying on the measurement of the antibodies or microorganism culture. Our first results showed that the PCR test can confirm the presence of Brucella in blood samples of infected patients. PMID- 9407548 TI - Tumor antigens discovery: perspectives for cancer therapy. AB - The adoptive transfer of cytotoxic T lymphocytes (CTLs) derived from tumor infiltrating lymphocytes (TIL) along with interleukin 2 (IL-2) into autologous patients with cancer resulted in the objective regression of tumor, indicating that these CTLs recognized cancer rejection antigens on tumor cells. To understand the molecular basis of T cell-mediated antitumor immunity, several groups started to search for such tumor antigens in melanoma as well as in other types of cancers. This led to the subject I will review in this article. A number of tumor antigens were isolated by the use of cDNA expression systems and biochemical approaches. These tumor antigens could be classified into several categories: tissue-specific differentiation antigens, tumor-specific shared antigens, and tumor-specific unique antigens. However, the majority of tumor antigens identified to date are nonmutated, self proteins. This raises important questions regarding the mechanism of antitumor activity and autoimmune disease. The identification of human tumor rejection antigens provides new opportunities for the development of therapeutic strategies against cancer. This review will summarize the current status and progress toward identifying human tumor antigens and their potential applications to cancer treatment. PMID- 9407550 TI - Molecular characterization of a mouse genomic element mobilized by advanced glycation endproduct modified-DNA (AGE-DNA). AB - BACKGROUND: DNA modified by advanced glycation endproducts (AGEs) undergoes a high frequency of insertional mutagenesis. In mouse lymphoid cells, these mutations are due in part to the transposition of host genomic elements that contain a DNA region homologous to the Alu family of repetitive elements. One particular 853 bp insertion, designated INS-1, was identified previously as a DNA element common to plasmids recovered from multiple, independent lymphoid cell transfections. MATERIALS AND METHODS: To characterize the genomic origin of this element, we used a 281-bp region of non-Alu-containing INS-1 sequence, designated. CORE, as a probe in Southern hybridization and for screening a bacteriophage mouse genomic DNA library. The resultant clones were sequenced and localized within the mouse genome. RESULTS: Two distinct genomic clones of 15 kB and 17 kB in size were isolated. A 522-bp unique region common to INS-1 and corresponding to the CORE sequence was identified in each clone. In both cases, CORE was found to be surrounded by repetitive DNA sequences: a 339-bp MT repeat at the 5' end, and a 150-bp B1 repeat at the 3' end. The CORE sequence was localized to mouse chromosome 1. CONCLUSIONS: These studies revealed that the CORE region of INS is present in low copy number but is associated with known repetitive DNA elements. The presence of these repetitive elements may facilitate the transposition of CORE by recombination or other, more complex rearrangement events, and explain in part the origin of AGE-induced insertional mutations. PMID- 9407551 TI - Caspase-mediated apoptosis in neuronal excitotoxicity triggered by nitric oxide. AB - BACKGROUND: Excitotoxicity and excess generation of nitric oxide (NO) are believed to be fundamental mechanisms in many acute and chronic neurodegenerative disorders. Disturbance of Ca2+ homeostasis and protein nitration/nitrosylation are key features in such conditions. Recently, a family of proteases collectively known as caspases has been implicated as common executor of a variety of death signals. In addition, overactivation of poly-(ADP-ribose) polymerase (PARP) has been observed in neuronal excitotoxicity. We therefore designed this study to investigate whether triggering of caspase activity and/or activation of PARP played a role in cerebellar granule cell (CGC) apoptosis elicited by peroxynitrite (ONOO-) or NO donors. MATERIALS AND METHODS: CGC from wild-type or PARP -/- mice were exposed to various nitric oxide donors. Caspase activation and its implications for membrane alterations, Ca2+ homeostasis, intracellular proteolysis, chromatin degradation, and cell death were investigated. RESULTS: CGC exposed to NO donors undergo apoptosis, which is mediated by excess synaptic release of excitotoxic mediators. This excitotoxic mechanism differs from direct NO toxicity in some other neuronal populations and does not involve PARP activation. Inhibition of caspases with different peptide substrates prevented cell death and the related features, including intracellular proteolysis, chromatin breakdown, and translocation of phosphatidylserine to the outer surface of the cell membrane. Increased Ca2+ influx following N-methyl-D-aspartate (NMDA) receptor (NMDA-R) activation was not inhibited by caspase inhibitors. CONCLUSIONS: In CGC, NO donors elicit apoptosis by a mechanism involving excitotoxic mediators, Ca2+ overload, and subsequent activation of caspases. PMID- 9407553 TI - Histone H2A significantly enhances in vitro DNA transfection. AB - BACKGROUND: Gene transfer is a potential treatment modality of genetic disease. Efficient, practical methods of DNA transfection are currently under investigation. MATERIALS AND METHODS: A beta-galactosidase reporter plasmid interacted electrostatically with histones, poly-L-Lys, poly-L-Arg, and a combination of poly-L-Lys and poly-L-Arg. This complex was then used to transfect COS-7 cells. beta-galactosidase activity was quantified and used to compare the efficiency of gene transfection in vitro. A comparison was also made of DNA transfection with the most active histone subclass, i.e., histone H2A, in the absence and presence of an anionic liposome. RESULTS: There was a marked increase in DNA transfection in the presence of histone H2A when compared with the control, whereas each of the other histones and polycations showed little, if any, effect. The extent of activation depends strongly on the DNA/histone ratio and is also a function of the molarity of the final Tris-acetate, pH 8, solution. The anionic liposomes used demonstrated an inhibitory effect. CONCLUSIONS: Histone H2A significantly enhances in vitro DNA transfection whereas other histones and anionic liposomes do not. A study of the difference between histone H2A and other histone subclasses may serve to clarify some of the mechanisms and the essential components of efficient gene delivery. PMID- 9407554 TI - Assessment of possible protective roles of selenium, zinc, and cis-stilbene oxide against acute T-2 toxin poisoning: a preliminary report. AB - The efficacy of two free radical scavengers, selenium and zinc, and a microsomal epoxide hydrolase-inducing agent, cis-stilbene oxide on the acute toxicity of T-2 toxin, a potent cytotoxic trichothecene, was investigated. Mice were pretreated daily for 3 consecutive days with either zinc sulfate (4.4 mg/kg, intraperitoneally [i.p.]), sodium selenite (1, 2, and 3 mg/kg i.p.) or cis stilbene oxide (50 mg/kg i.p.). A full 24-hr after the final dosing with these agents, mice were given T-2 toxin (2, 2.5, or 3 mg/kg i.p.). The acute lethal toxicity of T-2 toxin (2.5 mg/kg) was reduced by administration of only sodium selenite (3 mg/kg) and cis-stilbene oxide (50 mg/kg). No significant effect on weight gain was observed. PMID- 9407552 TI - Expression of TNF and TNF receptors (p55 and p75) in the rat brain after focal cerebral ischemia. AB - Cerebral ischemia induces a rapid and dramatic up-regulation of tumor necrosis factor (TNF) protein and mRNA, but the cellular sources of TNF in the ischemic brain have not been defined. The diverse activities of TNF are mediated via ligand interaction with two distinct receptors, p55 and p75, which activate separate intracellular signal transduction pathways, leading to distinct biological effects. Since the effects of cerebral ischemia on TNF receptor (TNFR) expression are unknown, we examined the cellular localization and protein expression of TNF and its two receptors in the rat cerebral cortex in response to permanent middle cerebral artery (MCA) occlusion. The results indicate that focal. cerebral ischemia up-regulates expression of TNF and both TNFRs within the ischemic cortex. The most abundant type of TNF immunoreactivity (IR) was a punctate and filamentous pattern of transected cellular processes; however, cell bodies of neurons, astrocytes, and microglia, as well as infiltrating polymorphonuclear (PMN) leukocytes also showed TNF IR. Brain vasculature displayed TNF IR not only within endothelial cells but also in the perivascular space. MCA occlusion induced significant up-regulation of TNF receptors, with p55 IR appearing within 6 hr, significantly before the appearance of p75 IR at 24 hr after the onset of ischemia. Since p55 has been implicated in transducing cytotoxic signalling of TNF, these results support the proposed injurious role of excessive TNF produced during the acute response to cerebral ischemia. PMID- 9407555 TI - Pacific ciguatoxin-1 associated with a large common-source outbreak of ciguatera in east Arnhem Land, Australia. AB - We report a retrospective study of the clinical signs and symptoms associated with a point-source outbreak of fish poisoning that occurred with a fish captured from the Arafura Sea, northern Australia. Twenty cases (16 Aboriginal and 4 non Aboriginal) characteristic of ciguatera, including 4 inpatients and 16 outpatients from the Gove Hospital, were identified based on the pattern of clinical symptoms and signs after ingestion of a large coral cod from a known ciguatera-prone coral reef. In the absence of a serologic test for the victim, laboratory analysis of a 230-g sample of the coral cod (Cephalopolis miniatus), using both mouse bioassay and HPLC/mass spectometry, showed that Pacific ciguatoxin-1 was the principal toxin involved. Intravenous mannitol was administered to one patient without clear benefit. Risk factors for ciguatera poisoning are ingestion of larger portions of reef fish from ciguatera-prone areas. Despite apparent local awareness of the distribution and etiology of the disease, large common-source outbreaks of ciguatera still occur. PMID- 9407556 TI - Apoptotic cellular damage in mice after T-2 toxin-induced acute toxicosis. AB - By histopathologic, electron microscopic, and immunochemical observation, the mechanism of cellular death was investigated in thymus, spleen, and liver of mice given intraperitoneally sublethal doses of T-2 toxin, a trichothecene mycotoxin. In the thymus and spleen of mice given 5.0 mg/kg body weight of T-2 toxin and killed 12 hours later, a massive cellular destruction characterized by chromatin condensation was evident, and electron microscopy analysis revealed the presence of apoptotic bodies. In the liver of mice given 2.5 mg/kg of T-2 toxin and killed 2 hours later, the induction of apoptotic cellular lesions was observed by electron microscopy, and Kupffer cells phagocytosed the apoptotic bodies. Such lesions were not observed in the mice killed 12 hours after receiving the toxin. In situ nick translation analysis (Tunel method) revealed DNA fragmentation in thymus, spleen, and liver shortly after administration of T-2 toxin. As previously observed in vitro, these findings indicated that T-2 toxin is a potent inducer of apoptotic cell death in thymus, spleen, and liver in vivo; especially in liver, apoptosis is induced rapidly as compared with the other tissues observed, and Kupffer cells play an important role for clearance of apoptosis. PMID- 9407557 TI - New saxitoxin analogues from the freshwater filamentous cyanobacterium Lyngbya wollei. AB - Along with decarbamoylsaxitoxin and decarbamoylgonyautoxin-2 and -3, six new saxitoxin analogues were isolated from the freshwater mat-forming filamentous cyanobacterium Lyngbya wollei collected from Guntersville Reservoir on the Tennessee River in Alabama. Their structures were determined by electrospray ionization mass spectrometry and several NMR techniques. Five of the toxins contain an acetyl moiety attached to the side chain, which is the first report of these saxitoxin analogues. In three of the toxins a hydrated ketone at C-12 was reduced to alpha-alcohol. The presence of acetate in the side chain resulted in a sevenfold to 17-fold times decrease in mouse toxicity compared to their carbamoyl counterparts, while the reduction at C-12 resulted in a complete loss of mouse toxicity. PMID- 9407558 TI - Effects of multiple doses of T-2 toxin on the erythroid response capacity of mice following an extensive experimental bleeding. AB - Total nucleated cellularity and total erythroid cell populations were measured in spleen and bone marrow of mice at different times after treatment with 3 daily doses of T-2 toxin (2.0 mg/kg). It was found that the initial depletion of hematopoietic cells produced by the toxin was rapidly reverted in spleen, giving way after 48 hr to a significant hypercellularity which after 10 days was 2.5 times the normal levels, but this effect was not observed in bone marrow, which slowly recovered normal cellularity after 5 days. The cytological analysis revealed that there was a highly significant shift in the ratio of erythroid to non-erythroid cells, since erythroid cell populations increased by about 8-fold in spleen and nearly 2-fold in bone marrow between 10 and 35 days after intoxication. In order to test the integrity of the hematopoietic reserve capacity, a hemorrhagic stress was produced in intoxicated animals at 10-50 days after toxin exposure. It was found that the erythroid response capacity was significantly higher in the intoxicated animals compared to anemic controls. The results suggest that the initial cytotoxic damage produced by T-2 toxin in the hematopoietic system is followed by a significant erythroid hypercellularity, which can confer an increased capacity for response to a hemorrhagic emergency. PMID- 9407559 TI - Antitumor effect of plant lectins. AB - This review examines the literature data concerning the biological activity of plant lectins. Numerous studies have reported that these substances possess toxic, cytotoxic, antitumor, and anticarcinogenic properties. A brief description of the biological properties of plant lectins, as well as the effect of plant lectins on normal and malignant cells and the antitumor properties of these lectins in vivo and in vitro, are included. These findings are interpreted and possible mechanisms of the antitumor effect of plant lectins are discussed. PMID- 9407560 TI - Identification of Protoceratium reticulatum as the biogenetic origin of yessotoxin. AB - Yessotoxin (YTX), a disulfated polyether toxin, was isolated from cultured cells of the marine dinoflagellate Protoceratium reticulatum and unambiguously identified by high-performance liquid chromatography, 1H NMR, and MS data. The result is the first to confirm toxigenicity of this species and demonstrate it as one of the biogenetic origins of YTX found in shellfish. PMID- 9407561 TI - Aflatoxin exposure is higher in vegetarians than nonvegetarians in Thailand. AB - Aflatoxin-albumin (AFB-albumin) adducts and hepatitis B markers (anti-HBs, and anti-HBc) were measured in vegetarians and nonvegetarians from Chiang Mai, Thailand. The AFB-albumin adduct levels were detected in 62% (37 of 60) of the vegetarian samples and 22% (22 of 100) of nonvegetarians. Somewhat higher levels were detected in vegetarians sera collected in the summer than in the winter, although this difference was not statistically significant. Subjects who were hepatitis B surface antigen (HBsAg)-positive had slightly higher AFB-albumin adduct levels than subjects who had evidence of past exposure (anti-HBc-positive) or no HB virus infection. This study indicated that vegetarians may have a higher frequency of aflatoxin exposure than nonvegetarians. Thai vegetarians consume various vegetables, grains, peanut, soybean, and fermented products, which have been reported to be sources of aflatoxin. PMID- 9407563 TI - Exhaled nitric oxide measurements in normal and asthmatic children. AB - The aim of this study was to determine whether we could measure exhaled nitric oxide (NO) levels in children, and whether the same pattern of exhaled NO concentrations was observed in asthmatic and normal children as had been seen in adults. Using a chemiluminescence NO analyzer, we measured NO in exhaled air both directly and through a T-piece allowing us to measure carbon dioxide (CO2), mouth pressure, and expiratory flows. In 39 normal children the mean peak exhaled NO was 49.6 parts per billion (ppb) (SD 37.4) when all expired gas passed directly through the NO analyzer, and 29.7 ppb (SD 27.1) when expiration occurred through a T-piece. The results were significantly higher in 15 asthmatic subjects on bronchodilator therapy only [126.1 ppb (SD 77.1) direct (P < 0.001), and 109.5 ppb (SD 106.8) via T-piece (P < 0.001)]. In 16 asthmatics on regular inhaled corticosteroids the mean peak exhaled levels were significantly lower 48.7 ppb (SD 43.3) direct (P < 0.001) and 45.2 ppb (SD 45.9) via T-piece (P < 0.01). There was no difference between the normal children and the asthmatic children on regular inhaled corticosteroids (P = 0.9 direct, P = 0.2 via T-piece). There were no significant differences in carbon dioxide levels, mouth pressure, duration of expiration and expiratory flows between the different groups, and no difference between carbon dioxide levels, mouth pressure and duration of expiration between the two methods (direct and T-piece). In 6 asthmatic children mean peak exhaled levels on NO fell from a median peak level of 124.5 ppb to 48.6 ppb when measured before and 2 weeks after commencement of inhaled corticosteroid treatment. The measurement of exhaled NO levels may be useful as a noninvasive means of monitoring children with asthma. PMID- 9407562 TI - Comparison of exhaled nitric oxide, serum eosinophilic cationic protein, and soluble interleukin-2 receptor in exacerbations of pediatric asthma. AB - The hypotheses tested in this study were that during acute asthma exacerbations (1) exhaled nitric oxide concentrations [eNO] are a more sensitive, noninvasive indicator of asthma disease activity than serum markers of inflammation such as eosinophil cationic protein (ECP) or soluble interleukin 2 receptor (sIL2R), and (2) elevated [eNO] are reduced after treatment with glucocorticoids (GC). Peak eNO levels were measured by chemiluminescence during slow expiration. Seven asthmatic subjects (mean age 11 yrs; mean morning FEV1 65% predicted) receiving inhaled GC, and with no radiographic evidence of acute sinusitis, were studied before and after a course of oral GC. Measurements of [eNO], ECP and sIL2R levels, and FEV1% were obtained before and after a course of GC. Six atopic nonasthmatic subjects (mean age 12 years; mean FEV1 94% predicted) and seven normal subjects (mean age 13 years; mean FEV1 100% predicted) were studied. The mean peak [eNO] level (parts per billion: ppb) for the asthma subjects before treatment (52 +/- 5 ppb SEM) was greater than the value for both nonasthmatic atopic and normal subjects (16 +/- 2 ppb and 14 +/- 2 ppb SEM, respectively; P < 0.0001). There was no significant difference in ECP or sIL2R values between asthmatic subjects and either atopic or normal subjects (P > 0.05). Baseline pre GC treatment ECP levels in the asthmatic subjects were significantly higher (P < 0.002) than post-GC treatment values. The mean peak [eNO] level in the asthmatic subjects declined after oral GC treatment to 14 +/- 1 ppb (P < 0.0002) and was less than 2 ppb different from either control group (P > 0.75). We conclude that [eNO] is a more sensitive marker of asthma disease activity than ECP and sIL2R levels. In addition, [eNO] appears to be a more useful indicator of the beneficial response to GC therapy than these other measurements in pediatric asthma. PMID- 9407564 TI - Response of premature infants with severe respiratory failure to inhaled nitric oxide. Preemie NO Collaborative Group. AB - Elevated pulmonary vascular resistance is seen in premature infants with severe respiratory distress syndrome (RDS). Inhaled nitric oxide (NO) has been shown to decrease pulmonary vascular resistance and to improve oxygenation in some patients with respiratory failure. The purpose of this study was to determine whether premature infants with severe RDS would respond to inhaled NO with an improvement in oxygenation. Eleven premature infants (mean gestational age 29.8 weeks) with severe respiratory failure caused by RDS were treated with NO in four concentrations [1, 5, 10, 20 parts per million (ppm) NO] and with placebo (0 ppm NO). Arterial blood gas measurements were drawn immediately before and at the end of each of the 15-minute treatments and were used to determine the arterial/alveolar oxygen ratio (PaO2/PAO2). Ten of the 11 infants had a greater than 25% increase in PaO2/PAO2. Five of the 11 had a greater than 50% increase in PaO2/PAO2. Despite normal cranial ultrasound imaging prior to NO, 3 infants had intracranial hemorrhage (ICH) noted on their first ultrasound scan after this brief period of NO treatment, and 4 additional infants developed ICH later during their hospitalization. No infant had significant elevations of methemoglobin concentrations after the total 60-minute exposure to NO. NO may be an effective method of improving oxygenation in infants with severe RDS. The disturbing incidence of ICH in this small group of infants needs to be carefully evaluated before considering routine use or NO for preterm infants. PMID- 9407565 TI - Pulmonary diseases in children with severe combined immune deficiency and DiGeorge syndrome. AB - Pulmonary disease is a common presenting feature and complication of T-cell immunodeficiency. We retrospectively reviewed 15 children with severe combined immune deficiency (SCID) and 19 children with DiGeorge syndrome at the time of their first presentation to the Royal Children's Hospital in the 15-year period from 1981 to 1995. In children with SCID, pulmonary disease was a common (67%) presenting feature and the organisms identified were Pneumocystis carinii (PCP) (n = 7), bacteria (n = 4), viruses (n = 3), and a fungus (n = 1). Late pulmonary complications included lower respiratory tract infections, bronchiolitis obliterans, and lymphointerstitial pneumonitis. Pulmonary infections were common (17 occasions) and the organisms identified were bacteria (n = 7), viruses (n = 6), fungi (n = 3), and Mycobacterium tuberculosis (n = 1). Pulmonary complications were responsible for 5 of 9 deaths. PCP was not identified as a late complication in any child, presumably as a result of effective prophylactic therapy. Although pulmonary disease was not a major presenting feature in children with DiGeorge syndrome, pulmonary complications were common. These included recurrent bacterial and viral infections and bronchomalacia, which complicated management and predisposed to morbidity and mortality, even in those without a T-cell defect. We conclude that pulmonary disease is a common manifestation in children with SCID and DiGeorge syndrome. PMID- 9407566 TI - Elevation of interleukin-8 and interleukin-6 precedes the influx of neutrophils in tracheal aspirates from preterm infants who develop bronchopulmonary dysplasia. AB - The influx of inflammatory mediators and cells into the tracheobronchial effluent of preterm infants with respiratory distress syndrome (RDS) appears to be important in signaling the development of bronchopulmonary dysplasia (BPD). The mechanism that initiates this early inflammatory response is not well understood. The purpose of this study was to test the hypothesis whether increased interleukin-8 (IL-8), a potent chemoattractant for human neutrophils, appears in the airways of preterm infants with RDS in whom BPD develops before the influx of neutrophils. In addition, airway secretions were analyzed for the cytokine interleukin-6 (IL-6) to test the hypothesis whether this pro-inflammatory cytokine is an early marker of inflammation in preterm infants with RDS who progress to BPD. Sixty-five infants less than 32 weeks gestation with RDS were enrolled on the first day of life and 56 infants completed the study, with 31 recovering from RDS (Non-BPD) and 25 infants progressing to BPD. Infants were excluded from enrollment in the presence of maternal chorioamnionitis, infection at birth, or infection within the first week of life. There were no significant differences in birthweight, gestational age, or prolonged rupture of membranes between the two groups. Serial tracheal aspirates (TA) were collected on days 1, 3, 5, and 7 while the infants remained intubated. Significant elevations of TA neutrophil counts were detected in the BPD group on days 5 and 7. Cell-free TA revealed marked elevations of IL-8 in the BPD group compared to the Non-BPD group [median (25th percentile, 75th percentile), ng/ml epithelial lining fluid (ELF)] on day 1 [BPD 485 (195, 840); Non-BPD 63.1 (28.3, 197), P < 0.05] and day 3 [BPD 740 (319, 1310); Non-BPD 111 (54.3, 337); P < 0.05], while on days 5 and 7, the differences were not statistically significant. Interleukin-6 (IL-6) was measured as a marker of acute inflammation and was not different in the two groups on day 1, but was significantly elevated on day 3 [median (25th percentile, 75th percentile), ng/ml ELF; BPD 297 (62.1, 702); Non-BPD 72 (32.8, 266), P < 0.05] and on day 5 [BPD 270 (136, 672); Non-BPD 86.4 (57.8, 138), P < 0.05]. These studies demonstrate that elevation of IL-8 and IL-6 levels precedes the marked neutrophil influx seen in the TA of preterm infants in whom BPD develop. The presence of IL-8 and IL-6 in TA from these infants suggests that these cytokines either initiate the acute inflammatory cascade in the lungs, or they are early markers of the inflammatory process that places preterm infants at high risk for BPD. PMID- 9407568 TI - Evaluation of predictors of weaning from mechanical ventilation in pediatric patients. AB - Objective criteria for ending mechanical ventilation have not been established for infants and children. A recent study in adult patients developed two new indexes, the Rapid Shallow Breathing Index (RSB) and the CROP Index for predicting success or failure of extubation. We decided to evaluate the applicability of these indices to intubated, mechanically ventilated pediatric patients. For this evaluation the indices were adapted to the physiology of infants and children. A pneumotachograph was used to measure spontaneous tidal volume, respiratory rate and dynamic compliance. The tidal volume and the dynamic compliance were corrected for the patient's body weight. Based on the data collected a cutoff value for each index was determined. Of 47 sets of patient data, 38 (81%) were collected during successful extubations, 9 (19%) during failed extubations. A modified CROP index value of > or = 0.1 ml x mmHg/bpm/kg and a modified RSB index value of < or = 11 bpm/mlkg were identified as predictive of successful extubation. The modified CROP cutoff value produced a sensitivity and specificity of 1.0; respective values for the modified RSB cutoff value are 0.79 and 0.78. Cutoff values of > or = 0.1 and < or = 11 for the modified CROP index and RSB index, respectively, appear to be predictive of successful extubation in the pediatric population. Our data identifies the modified CROP index as a superior discriminator between successful and unsuccessful extubation. PMID- 9407567 TI - Surfactant replacement therapy improves ventilation inhomogeneity in infants with respiratory distress syndrome. AB - Surfactant deficiency in newborn infants with hyaline membrane disease (HMD) reduces peripheral airway stability, leading to lung atelectasis, inhomogeneity of distribution of ventilation, ventilation/perfusion mismatch, and hypoxemia. The aim of this study was to evaluate the immediate effect of exogenous surfactant treatment on ventilation inhomogeneity (VIH) in infants with HMD. Homogeneity of ventilation was measured repeatedly in ten infants (median gestational age 30 weeks and birthweight 1.50 kg) after Exosurf, and in six infants (median gestational age 30 weeks and birthweight 1.42 kg) after Survanta treatment. Lung function was measured before and 0.5, 2, and 6 hours after administration of a single dose of surfactant. The multiple breath nitrogen washout method was used to measure the time pattern of nitrogen elimination from the lungs. VIH was evaluated by using both a compartmental lung model and a model independent moment analysis. The two-compartment lung model was found to dominate before surfactant treatment, while a single-compartment model (implying homogeneous ventilation) fitted the washout data best 6 hours after Exosurf treatment (P < 0.01). The same pattern occurred 2 hours after Survanta administration. Moment analysis confirmed the reduction in VIH by both surfactants. This study supports the hypothesis that the improved oxygenation after surfactant treatment in infants with HMD results from a reduction in VIH and an increase in functional residual capacity (FRC). PMID- 9407569 TI - Frequency responses of infant air-balloon versus liquid-filled catheters for intra-esophageal pressure measurement. AB - Amplitude and phase frequency response characteristics of infant air-balloon catheters (IABC) of differing French gauge (FG) sizes and brands were quantified to determine their suitability for measuring dynamic intra-esophageal pressure (Pes) accurately. Frequency response performances of matching IABC and water filled catheters (WFC) were also compared using the swept sine wave technique. The maximum respiratory rate within which IABCs could potentially measure Pes within a 5% error limit was calculated (FRR). Frequency responses of IABCs greater than FG size 5 exhibited underdamped resonant properties, while smaller FG size IABCs exhibited near-critical damping or overdamping. IABCs maintained uniform amplitude frequency responses up to 25 Hz, demonstrating the ability to measure Pes potentially up to 148 breaths/min within a 5% error limit. The frequency response performance of FG size 6 IABCs was similar to that of FG size 10 IABCs. Compared with matching WFCs, the frequency response performance of IABCs was significantly superior, the frequency response variability within IABC samples was lower, and IABC correlation between FG size and FRR was advantageously lower than for WFCs. FRR values for differing IABC brands and FG sizes are presented. We conclude that IABCs manufactured to infant-appropriate balloon specifications exhibit significantly superior frequency response characteristics compared with matching WFCs. Measurement accuracy is not improved using IABCs greater than FG size 6. Inexpensive intra-esophageal IABCs are technical suitable for the accurate measurement of dynamic Pes during high frequency respiratory mechanics encountered during infant artificial ventilation. PMID- 9407570 TI - Severe laryngomalacia and bronchomalacia in DiGeorge syndrome and CHARGE association. PMID- 9407571 TI - Surfactant therapy improves pulmonary function in infants with Pneumocystis carinii pneumonia and acquired immunodeficiency syndrome. AB - Pneumocystis carinii pneumonia (PCP) is an important cause of acute respiratory failure in HIV-infected children. PCP may initiate acute respiratory distress syndrome (ARDS) by adversely affecting surfactant physiology. We report improved pulmonary function following administration of bovine lipid extract surfactant to two infants with AIDS-related PCP/ARDS. PMID- 9407572 TI - Consultation-liaison psychiatry research: more like a ground cover than a hedgerow. PMID- 9407573 TI - Psychosocial factors in fibromyalgia compared with rheumatoid arthritis: I. Psychiatric diagnoses and functional disability. AB - OBJECTIVE: Recent studies of the relationship between fibromyalgia and psychiatric disorders have yielded conflicting findings, and many of these inconsistencies seem to result from methodological differences. METHOD: We compared 36 patients with fibromyalgia and 33 patients with rheumatoid arthritis from a tertiary care clinic using physician-administered, structured psychiatric interviews and self-reported measures of illness appraisal, coping, and functional disability. RESULTS: Patients with fibromyalgia had significantly higher lifetime prevalence rates of mood and anxiety disorders, as well as higher mean numbers of medically unexplained physical symptoms across several organ systems. Ninety percent of the patients with fibromyalgia had a prior psychiatric diagnosis compared with less than half of the patients with rheumatoid arthritis. CONCLUSIONS: Despite the absence of organic pathology, the patients with fibromyalgia had equal or greater functional disability and were less well adapted to their illness. Although the pathophysiology of fibromyalgia remains unclear, co-morbid psychiatric disorders and functional disability remain an important focus of treatment in this population. PMID- 9407574 TI - Psychosocial factors in fibromyalgia compared with rheumatoid arthritis: II. Sexual, physical, and emotional abuse and neglect. AB - OBJECTIVE: Two recent reports have found associations between fibromyalgia and sexual victimization, but had methodologic characteristics that limited their interpretation. METHOD: We compared 36 patients with fibromyalgia and 33 patients with rheumatoid arthritis by using structured interviews for sexual, physical, and emotional victimization histories, as well as dimensional self-report measures of victimization severity. RESULTS: Compared with the patients with rheumatoid arthritis, those with fibromyalgia had significantly higher lifetime prevalence rates of all forms of victimization, both adult and childhood, as well as combinations of adult and childhood trauma. Although childhood maltreatment was found to be a general risk factor for fibromyalgia, particular forms of maltreatment (eg, sexual abuse per se) did not have specific effects. Experiences of physical assault in adulthood, however, showed a strong and specific relationship with unexplained pain. Trauma severity was correlated significantly with measures of physical disability, psychiatric distress, illness adjustment, personality, and quality of sleep in patients with fibromyalgia but not in those with rheumatoid arthritis. CONCLUSIONS: Fibromyalgia seems to be associated with increased risk of victimization, particularly adult physical abuse. Sexual, physical, and emotional trauma may be important factors in the development and maintenance of this disorder and its associated disability in many patients. PMID- 9407575 TI - Long-term outcome of motor vehicle accident injury. AB - OBJECTIVE: To define the psychological outcome at 5 years of a sample of non-head injured motor vehicle accident victims and identify baseline predictors. METHODS: Self-report questionnaires were completed by 111 consecutive subjects who had been injured in a motor vehicle accident 5 years earlier and who had been assessed previously in a prospective 1-year study. RESULTS: Although most subjects reported a good outcome, a substantial minority described continuing social, physical, and psychological difficulties and a quarter of those studied suffered phobic anxiety about travel as a driver or passenger. There was little change in quality of life outcome and effects on travel between assessments at 3 months, 1 year, and 5 years. The prevalence of posttraumatic stress disorder remained approximately 10% throughout the follow-up; most early cases had remitted by 5 years, and a similar number of delayed new onsets had occurred between 1 year and 5 years. PTSD at 5 years was predicted by physical outcome and by postaccident intrusive memories and emotional distress. Compensation proceedings were initiated by 66 subjects and were often prolonged and a cause of distress. There were no significant associations with outcome. Trends for a poor outcome in claimants, especially those not settled at 5 years, may be due to their having more serious physical problems. CONCLUSION: Psychological complications are important and persistent after injury in a motor vehicle accident, are associated with adverse effects on everyday activities, and pose a challenge for consultation-liaison psychiatry. PMID- 9407576 TI - A randomized trial of geriatric liaison intervention in elderly medical inpatients. AB - OBJECTIVE: The aim of this study was to examine the effect of psychogeriatric intervention in a group of elderly medical inpatients over 75 years of age. In addition to usual care, intervention consisted of multidisciplinary joint treatment by a psychogeriatric team. The main purpose of intervention was to obtain the optimal level of physical functioning. METHOD: In a prospective randomized trial the effect of the intervention (N = 140) compared with usual care (N = 97) was estimated for physical functioning, length of stay, and nursing home placement within 12 months of discharge. RESULTS: Substantially more patients assigned to the intervention group improved in their physical functioning, and fewer became worse. The mean length of stay was 5 days shorter for the intervention group. There were more readmissions to hospital in the usual care group (29.9%) compared with the intervention group (17.4%). Of the patients assigned to the intervention treatment, 18% were admitted to a nursing home. In the usual care group this was 27%. The effects of intervention remained statistically significant for all the outcome variables after controlling for possible confounding baseline characteristics. CONCLUSIONS: The intervention we studied had clinically relevant effects on important outcome variables. Psychiatric co-morbidity was an important risk factor for the outcome of the patients in our study. By combining elements from a psychiatric and geriatric consultation service with elements from a unit-driven service, we were able to improve health care for the elderly in our hospital in a feasible and cost effective way. PMID- 9407577 TI - Do patients with "pure" chronic fatigue syndrome (neurasthenia) have abnormal sleep? AB - OBJECTIVE: To determine whether patients with "pure" chronic fatigue syndrome (neurasthenia) have sleep abnormalities which may contribute to subjective measures of daytime fatigue. METHOD: Sleep characteristics of 20 patients meeting research criteria for chronic fatigue syndrome (CFS) but not depression, anxiety, or sleep disorder were compared with sleep characteristics of 20 healthy subjects matched for age and sex. Measures of sleep included a) subjective interview reports and sleep diaries and b) home-based polysomnography. RESULTS: Patients with CFS complained of poor quality unrefreshing sleep. They also napped during the day. Polysomnograph data showed no difference in actual nocturnal sleep time between the two groups although patients with CFS spent significantly longer in bed (p < .01), slept less efficiently (p < .03), and spent longer awake after sleep onset (p < .05). The polysomnographs of seven patients with CFS and one healthy subject were regarded as significantly abnormal. Five patients and one healthy subject had difficulty maintaining sleep. One patient had a disorder of both initiating and maintaining sleep and one patient woke early. CONCLUSIONS: Patients with "pure" CFS complain of unrefreshing sleep but only a minority have a clearly abnormal polysomnograph. The most common abnormality is of long periods spent awake after initial sleep onset. Although sleep abnormalities may play a role in the etiology of CFS, they seem to be unlikely to be an important cause of daytime fatigue in the majority of patients. However, pharmacological and behavioral methods that improve sleep quality may be an important component of a pragmatically based treatment package for patients who do have abnormal sleep. PMID- 9407578 TI - Iatrogenic factors and chronic pain. AB - OBJECTIVE: Although it is accepted that the etiology of chronic pain is multifactorial, little attention has been given to the possible role of iatrogenesis. The aim of the present study is to identify possible iatrogenic factors in chronic pain patients. METHODS: We report a cross-sectional study of 125 patients attending specialist pain clinics in South London. Data were collected using semistructured checklists. Patients were interviewed with a structured psychiatric interview and were given a questionnaire booklet to complete. RESULTS: We found that possible iatrogenic factors, such as over investigation, inappropriate information and advice given to patients as well as misdiagnosis, over-treatment, and inappropriate prescription of medication were common in this sample. CONCLUSIONS: We suggest that future studies should take account of the role of the doctors, as well as that of the patients, in the etiology, and hence prevention of chronic pain. PMID- 9407579 TI - Diseases among men 20 years after exposure to severe stress: implications for clinical research and medical care. AB - OBJECTIVE: Epidemiologic studies have linked exposure to severe environmental stress, such as natural disasters and combat operations, to the onset of specific psychiatric disorders. Some research also suggests that these exposures may be associated with the onset of chronic diseases as well. However, these chronic disease outcome studies often have been obscured by bias and confounding. METHOD: The medical histories of 1399 male Vietnam veterans approximately 20 years after combat exposure (mean years = 17) were analyzed by lifetime posttraumatic stress disorder (PTSD) status (lifetime PTSD = 332 cases). These men were included in a national, random in-person study of United States Army veterans of the Vietnam War (study completion rate = 65%). RESULTS: After controlling for preservice, in service, and postservice factors (including intelligence, race, region of birth, enlistment status, volunteer status, Army marital status, Army medical profile, hypochondriasis, age, smoking history, substance abuse, education, and income), associations were found for reported circulatory [odds ratio (OR) = 1.62, p = .007], digestive (OR = 1.47, p = .036), musculoskeletal (OR = 1.78, p = .008), endocrine-nutritional-metabolic (OR = 1.58, p = .10), nervous system (OR = 2.47, p < .001), respiratory (OR = 1.54, p = .042), and nonsexually transmitted infectious diseases (OR = 2.14, p < .004) after military service. CONCLUSION: Although this study has some limitations, it suggests that there is a direct link between severe stress exposures and a broad spectrum of human diseases. In the future, medical researchers and clinicians should focus more on the medical consequences of exposure to severe environmental stress and seek to better integrate psychobiologic models of disease pathogenesis. PMID- 9407580 TI - Psychosomatic research at the margins of morality--war as a stressor. PMID- 9407581 TI - The influence of ambient light and menstrual status on the moods of a nonclinical population of young women. AB - OBJECTIVE: To study the influence of the daily variation in ambient light and menstrual status on mood fluctuation in a nonclinical population of young women. METHODS: Women kept mood diaries (two per day) over a period of 32 days that straddled the spring equinox. One group believed the purpose of the study was to investigate women's moods are significantly elevated by light, and this elevation occurs irrespective of the subjects knowledge of the experimental purpose. No evidence for a depression of women's mood in the premenstruum was found, although women who claimed to suffer from premenstrual syndrome (PMS) showed more reversals of their mood during the 32 days records were kept. CONCLUSIONS: The results highlight the fact that an individual's mood may be influenced by the levels of ambient light as well as the photoperiod. PMID- 9407582 TI - Active coping and cardiovascular reactivity: a multiplicity of influences. AB - OBJECTIVE: Active coping enhances cardiovascular response presumably by beta adrenergically mediated myocardial activation. This study examined impedance derived hemodynamic parameters underlying blood pressure response to two laboratory tasks requiring active coping, performed either with or without an appetitive (i.e., monetary) incentive. METHOD: Forty-eight healthy, young men completed the Stroop Color-Word Test and Mirror Tracing. Half received no incentive, whereas half were provided with a monetary incentive as an active coping manipulation. Task-related changes in blood pressure, heart rate, systolic time intervals, and hemodynamic parameters were monitored. Psychological responses to the tasks were also obtained. RESULTS: On average, incentive virtually doubled blood pressure response to both Stroop and Mirror Tracing. The change in blood pressure was explained predominantly by a concomitant increase in total peripheral resistance. Heart rate response was also enhanced substantially with incentive. Individuals in the incentive condition reported greater interest in the task, but less perceived control, than persons in the no-incentive condition. CONCLUSIONS: The incentive-related increase in total peripheral resistance, combined with an absence of enhanced stroke volume, cardiac output, or preejection period response, indicates that active coping may, under certain conditions, elevate blood pressure via increased systemic resistance, presumably reflecting alpha-adrenergic activation. Furthermore, the enhanced heart rate associated with incentive may reflect a withdrawal of parasympathetic influence. PMID- 9407583 TI - Personality predictors of dimensions of psychosocial adjustment after surgery. AB - OBJECTIVE: Although many studies have examined the relationship between personality factors and adjustment after surgery, most of them have had very short follow-up periods. The present prospective study examines whether preoperative psychodynamic assessment of personality traits enhances prediction of various areas of psychosocial adjustment assessed at least 1 year after surgery. METHODS: In 53 patients undergoing pelvic pouch surgery for ulcerative colitis, we examined the relationship between personality traits measured before surgery, and postoperative psychosocial adjustment assessed at a median of 17 months postoperatively, controlling for the effect of surgical functional outcome. Personality traits were assessed with the Karolinska Psychodynamic Profile (KAPP). Surgical functional outcome scales and the Psychosocial Adjustment to Illness Scale (PAIS) were used. RESULTS: Problems with sexual satisfaction, perfectionistic body ideals, lack of alexithymia, and poor frustration tolerance predicted poor postoperative adjustment in various areas, beyond what was predicted by surgical functional outcome alone. Moreover, moderate preoperative levels of alexithymia were beneficial to postoperative adjustment in the area of psychological distress. CONCLUSIONS: The findings suggest that the preoperative assessment of the patient's long-term sexual functioning and satisfaction, the importance attached to his or her appearance, level of alexithymia, and general capacity to tolerate frustration and set-backs in life, might alert both the surgeon and the patient to potential risk factors for poor postsurgical adjustment. PMID- 9407584 TI - Anger expression, age, and blood pressure in modernizing Samoan adults. AB - OBJECTIVE: Relationships among anger expression, age, and blood pressure (BP) were studied in a cross-sectional sample of 593 American and Western Samoan adult men and women, 25 to 55 years of age. Prior studies indicated that anger coping is an important psychosocial domain in modernizing Samoans. METHODS: Anger expression was assessed using a modified 24-item version of the State-Trait Anxiety Inventory composed of anger-in, anger-out, and anger-control, along with 4 Samoan culture-specific anger items. Age and sex stratified analyses were performed. Body-mass adjusted BP was regressed on the anger expression subscales and age. RESULTS: In women < or = 40 years of age, anger-out was significantly (p < 0.01) and negatively related to adjusted diastolic BP. Young women from American and Western Samoa who outwardly expressed anger least frequently had higher adjusted diastolic BP. CONCLUSION: The significant influence of anger expression on BP in young modernizing Samoan women may be because: a) increased stress from the interaction of traditional gender role-related domestic demands and more opportunities for individual socioeconomic activities; and b) the culturally normative pattern of suppressed emotional expression. PMID- 9407585 TI - Cognitive slowing in chronic fatigue syndrome (CFS) PMID- 9407586 TI - Heat shock-induced homeotic transformations of the axial skeleton and associated shifts of Hox gene expression domains in mouse embryos. AB - Pregnant ICR mice were immersed in water at 42 degrees C for up to 15 min on Day 8.5 of gestation (plug day = Day 0), and their term fetuses were double stained with alcian blue and alizarin red S for skeletal examination. Heat exposure for 15 min induced homeotic vertebral transformations in more than one-third of the living fetuses, in which the morphologic identity of vertebrae (T6-S1) was shifted anteriorly by one or two segmental levels. The frequency of fetuses with vertebral transformations and the degree of the shift of vertebral identity were dependent on the length of heat exposure. The expression domains of Hoxa-7, Hoxc 8, and Hoxc-9 genes as examined by whole mount in situ hybridization were found to be shifted anteriorly in heated embryos. The heat-induced shifts of Hox gene expression domains were consistent with the observed vertebral transformations and suggested correlation or colinearity with the clustered organization of the Hox genes. The result of the present study indicates that a brief heat shock at a critical stage of differentiation can interfere with the normal establishment of Hox codes and subsequently, perturb the specification of vertebral identity. PMID- 9407587 TI - In vivo and in vitro developmental toxicity in LPS-induced zinc-deficient rabbits. AB - Lipopolysaccharide (LPS) was used to induce maternal hypozincemia in order to test the hypothesis that altered zinc homeostasis is developmentally toxic in the rabbit. Treatment of dams on Gestation Day (GD) 8 with LPS (0.67 microgram/kg i.v.) caused total resorption of 78% (7 of 9) of the litters whereas GD 10 treatment increased the percentage of resorbed implantations (18-fold), but resulted in only 14% (1 of 7) totally resorbed litters. Cotreatment with zinc oxide (ZnO) on GD 10 decreased the resorption rate by 44%, indicating that hypozincemia was partially responsible for the resorptions. However, ZnO had no effect on resorption rate in GD 8 LPS-treated dams. No malformations were observed with LPS dosing on either gestation day. To determine whether LPS induced Zn deficiency had any direct effects on rabbit embryos, normal GD 9 embryos were cultured for 48 h in serum from LPS-treated dams (0.53 +/- 0.01 microgram/mL Zn) or from controls (1.74 +/- 0.07 micrograms/mL Zn). Embryo growth and development were normal in both groups, indicating a lack of any direct embryo effects of Zn deficiency. Finally, maternal plasma progesterone and the Zn content of conceptus tissues were measured 24 h after LPS injection on GD 10. Despite a marked decrease in maternal serum Zn, no significant changes in embryo, visceral yolk sac, yolk sac cavity-exoceolomic fluid, or placental Zn were found. However, maternal progesterone levels were decreased 33 and 28% in the LPS and LPS + ZnO groups, respectively. Taken together, these results indicate that rabbits may differ from rodent species in their lesser susceptibility to the teratogenic potential of transient maternal Zn deficiency, as well as in their resistance to conceptus Zn changes. Nonetheless, Zn deficiency may be responsible for an increase in resorption rate in the rabbit. PMID- 9407588 TI - Developmental toxicity of dietary zinc deficiency in New Zealand white rabbits. AB - Chemically induced maternal Zn deficiency has been shown previously to cause terata and increase embryonic loss in rodents. To examine the potential effects of Zn deficiency in the rabbit, a major developmental toxicity test species, rabbit dams were fed an ethylenediamine-tetraacetic acid-washed alfalfa-based Zn deficient diet (-Zn) or the same diet replete with 80 ppm Zn (control) from Gestation Day (GD) 0 through 20. A third group of animals was pair fed to match the mean daily feed consumption levels of the < 2 ppm Zn group. By GD 7, maternal serum Zn levels of the -Zn dams were decreased 56% and reached a nadir with a 75% decrease of serum Zn by GD 14. Zinc concentrations in the visceral yolk sac and visceral yolk sac-exoceolomic fluid were decreased 30% and 50%, respectively, by GD 11. Although GD 11 embryonic Zn levels were not affected, the embryos from Zn deficient dams exhibited decreased head length, somite number, and total protein. On GD 28, a significant increase in resorptions/litter was noted in the -Zn group, and the incidence of totally resorbed litters of the -Zn group was greater than laboratory historical control values. No terata were observed in GD 28 fetuses. This study indicates that Zn deficiency occurring during the standard dosing period of guideline rabbit developmental toxicity studies may be associated with a modest increase in resorption rate and a transient inhibition of embryonic growth, but in contrast to rodent species, does not appear to be teratogenic. PMID- 9407589 TI - Interactions between endosulfan and dieldrin on estrogen-mediated processes in vitro and in vivo. AB - There is growing concern that estrogenic chemicals, both natural and human-made, may be causing a variety of reproductive disorders in wildlife and human populations. Recent in vitro data suggest that the interaction between some weakly estrogenic organochlorines, dieldrin, endosulfan, toxaphene, and chlordane, causes a synergistic increase in their estrogenic potency, an effect due to joint action on estrogen receptors (ER). As these studies were conducted using models of estrogen action derived from cells that are not physiologically controlled by estrogens, the relevance of these findings to human health are not clear. The present studies were conducted to examine the interaction between endosulfan and dieldrin in the activation of ER in or extracted from mammalian cells. Endosulfan and dieldrin showed no synergism in displacing 3H-E2 from rat uterine ER or in inducing the proliferation of MCF-7 breast cancer cells, an estrogen-dependent response. Furthermore, endosulfan (0.1 mg per animal per d) or dieldrin (0.1 mg), alone or in combination, injected intraperitoneally daily for 3 d, did not stimulate any uterotrophic activity nor had any effect on pituitary prolactin or other endocrine-related endpoints in immature female rats. These studies demonstrate that these weakly estrogenic compounds do not interact in a synergistic fashion in binding to ER or in activating ER-dependent responses in mammalian tissues or cells. Thus, these results suggest that coexposure to these weakly estrogenic environmental contaminants likely will not cause human reproductive toxicity related to estrogen action. PMID- 9407590 TI - Effects of corticosterone on reproduction in male Sprague-Dawley rats. AB - Corticosterone, the predominant circulating adrenal corticosteroid in rodents, was investigated for its effects on reproduction in male Sprague-Dawley rats. Male rats (in groups of 50, 25, and 50) were administered corticosterone at doses of 0, 10, and 25 mg/kg/d, respectively, by subcutaneous injection once daily for 6 weeks; the highest dose was decreased to 20 mg/kg/d after 15 d. During the last 2 weeks of the 6-week treatment period, 25 males per group were paired with untreated females. The remaining 25 males from the 0 and 25/20 mg/kg/d groups were allowed a 6-week recovery period and, during the last 2 weeks of this period, these males were also paired with untreated females. At the end of the treatment period, the males had markedly elevated plasma corticosterone concentrations and decreased weight gain. They also produced fewer copulatory plugs than controls, which may have been secondary to observed adverse effects on the accessory sex organs (decreased weights and microscopic changes in prostate and seminal vesicles). However, no adverse effects on sperm motility, sperm count, or microscopic features of the testes were observed. Serum testosterone concentration of the high-dose males was elevated, but luteinizing hormone was unaffected. The numbers of embryonic implantation sites and live fetuses in females mated to these males were reduced. All of these effects except decreased prostate weights were reversible upon cessation of corticosterone administration. Thus, exogenous administration of corticosterone to male rats produced reversible effects on implant count and litter size of female rats mated to these males. These effects on male rat reproduction may have been secondary to reduced accessory sex organ function, which resulted in diminished secretions and fewer copulatory plugs. PMID- 9407591 TI - Decreased superovulation in adult mice following neonatal exposures to technical methoxychlor. AB - To examine the effects of technical methoxychlor (MXC) on superovulation, neonatal mice received intraperitoneal (i.p.) injections of either sesame oil, 10 micrograms of estradiol 17 beta, or 0.1, 0.5, or 1 mg of technical MXC. At 2 and 4 months, half of the mice received a superovulatory regimen of 10 IU pregnant mare's serum gonadotropin followed by 10 IU human chorionic gonadotropin. The mice were sacrificed 15 to 20 h later, the number of ovulated oocytes were counted, and the ovaries were removed for histology. In the lowest MXC dose, the ovaries appeared normal and at 2 months, ovulated the same number of oocytes as controls. Estradiol or the highest two MXC doses induced ovarian atrophy. Following gonadotropin injections, these ovaries also ovulated oocytes. However, the number of oocytes recovered from experimental mice exhibited a time- and dose dependent decline, and by 4 months, their number was significantly reduced. Neonatal exposures to MXC reduces ovulatory rates and ovarian functions in adults. PMID- 9407592 TI - Initial uterine alterations caused by developmental exposure to tamoxifen. AB - Neonatal treatment of rodents with the widely used antiestrogen tamoxifen causes endometrial cancer and reproductive tract lesions reminiscent of the diethylstilbestrol (DES) syndrome. To evaluate the initial alterations induced in the developing uterus by tamoxifen or DES, neonatal Sprague-Dawley rat pups received 100 micrograms of tamoxifen (Group 1), 1 microgram of DES (Group 2), or vehicle (Group 3) subcutaneously on days 1 through 5, and their uteri were studied by light microscopy, 5-bromo-2' deoxyuridine immunohistochemistry, and computer-based morphometry. At Postnatal Day 6, epithelial hypertrophy (184.3% and 237.9% of controls) and myometrial thickening (151.9% and 180.0%) accounted for the uterotrophic effects of tamoxifen and DES. Evidence of secretory activity in epithelial cells, reduction of the epithelial BrdU-labeling index to 18.1% (tamoxifen) and 41.1% (DES) of controls, premature endometrial and myometrial differentiation, and the presence of eosinophils in both treatment groups suggested that tamoxifen exerted a DES-like estrogenic action on the developing uterus. These findings indicate that immediate epithelial and stromal-myometrial uterine alterations are found at Postnatal Day 6 after neonatal tamoxifen treatment. PMID- 9407593 TI - Exacerbation of acetazolamide teratogenesis by amiloride and its analogs active against Na+/H+ exchangers and Na+ channels. AB - Postaxial forelimb ectrodactyly induced by acetazolamide given on Day 9.5 of murine gestation is thought to be mediated by reduced intracellular pH (pHi) within the limb bud. Coadministration of amiloride increases the incidence and severity of acetazolamide-induced forelimb malformations and further reduces limb bud pHi. These findings were hypothesized to be attributable to the action of amiloride as an inhibitor of Na+/H+ exchangers (NHEs), plasma membrane-localized proteins involved in the maintenance of cellular pH homeostasis. Here, we explored this hypothesis further by coadministering with acetazolamide, amiloride, or analogs known to preferentially inhibit NHEs 5-(N-methyl-N isobutyl)-amiloride, 5-(N, N-hexamethylene)-amiloride, 5-(N, N-dimethyl) amiloride, and 5-(N-ethyl-N-isopropyl)-amiloride or amiloride-sensitive Na+ channels (benzamil). The coadministration of either amiloride, benzamil, 5-(N, N dimethyl)-amiloride, 5-(N-ethyl-N-isopropyl)-amiloride, or 5-(N-methyl-N isobutyl)-amiloride all dose responsively increased the frequency and severity of forelimb malformations compared to acetazolamide alone. None of the analogs given alone induced forelimb ectrodactyly. The data are consistent with the original hypothesis that the exacerbation of acetazolamide teratogenesis is due to NHE inhibition. Surprisingly, benzamil was the most potent potentiator of acetazolamide teratogenesis. This result strongly suggests that amiloride sensitive Na+ channels are also present within the murine embryo and are likely to play a role in pHi homeostasis. PMID- 9407594 TI - Blood boron concentrations in pregnant rats fed boric acid throughout gestation. AB - Timed-mated Sprague-Dawley rats (28 to 32/group) were exposed to boric acid (BA) in the diet from Gestational Day (GD) 0 to 20. Dietary concentrations of added BA (0%, 0.025%, 0.050%, 0.075%, 0.100%, or 0.200%) yielded average daily intakes equivalent to 0, 3, 6, 10, 13, or 25 mg boron/kg body weight/d. Dams and their fetuses were evaluated for evidence of maternal or developmental toxicity, as reported previously. At termination on GD 20, maternal whole blood was collected in heparinized Vacutainer tubes, stored frozen (-20 degrees C), and subsequently prepared by a high-temperature alkaline ashing procedure for analysis of boron by inductively coupled plasma optical emission spectrometry. Increasing dietary concentrations of BA were positively associated with whole blood boron concentrations in confirmed pregnant rats, specifically 0.229 +/- 0.143, 0.564 +/ 0.211, 0.975 +/- 0.261, 1.27 +/- 0.298, 1.53 +/- 0.546, or 2.82 +/- 0.987 micrograms boron/g whole blood (mean +/- SD) for the control through high-dose groups. Maternal blood boron concentrations were positively correlated with indices of maternal dietary intake of boron and with embryo/fetal toxicity observed at 0.100% and 0.200% BA in the diet reported previously. Thus, blood boron concentrations of 1.27 +/- 0.298 and 1.53 +/- 0.546 micrograms boron/g were associated with the no-observed-adverse-effect level (10 mg boron/kg/d) and lowest-observed-adverse-effect level (13 mg boron/kg/d) for developmental toxicity reported previously. PMID- 9407595 TI - Retinoic acid-induced asymmetric craniofacial growth and cleft palate in the TO mouse fetus. AB - The etiology and pathogenetic mechanisms of cleft palate (CP) are rather uncertain. Both genetic and environmental factors are known to cause failure of horizontalization and/or failure of fusion of the palatal shelves resulting in CP. Retinoic acid (RA)-induced CP in the mouse is reported to exhibit two peaks of incidence separated by a less sensitive window. The morphologic bases of the differential sensitivity are not known. The objectives of this study were to determine whether the TO mouse had similar peaks of sensitivity to RA-induced CP, and if it did, to evaluate the morphologic and histologic bases of CP induced at an early [Gestation Day (GD) 8] and at a late (GD 12) stage of embryonic development. Single doses of all-trans-RA were administered to groups of mice on one of GD 8 to 15. On GD 18, fetuses were evaluated for the presence of CP, and the developmental stage of the palatal shelves was determined. All doses of RA were found to induce a high incidence of CP in the GD 8 to 13 treatment groups. GD 14 and 15 were not susceptible. There were no stage-dependent peaks or less sensitive windows, indicating that RA-induced CP in this strain is a continuum from GD 8 through 13. Morphologically clefting in the GD 8-RA treatment group was characterized by extreme hypoplasia (65% to 100%, depending on the dose) or agenesis (35% in the 200 mg/kg group) of the palatal shelves and associated with astomia, microstomia, aglossia, microglossia, and micrognathia with fusion of mandible, maxilla, and zygoma. Treatment on subsequent days of gestation resulted in CP with the shelves reaching progressively higher levels of maturity in terms of developmental staging. There was no case of CP with horizontalized shelves apposing but failing to fuse with each other. The facial skeleton of GD 12-RA group was hypoplastic but not malformed. Reduction in all dimensions of the cranium and mandible was highly significant (P < 0.001) in the GD 8-RA group, whereas there was a clear imbalance between the vertical growth and that in other directions in the GD 12-RA group. The CR length, head and body weights, and the protein content of heads of GD 8-RA-treated embryos were significantly reduced. Histologic studies showed that both the intrinsic and extrinsic muscles of the tongue and face, growth of the Meckel's cartilage, and ossification of the mandible were severely affected in the GD 8 treatment group, whereas these tissues were only moderately affected in the embryos of the GD 12-RA group. However, the quality of cytodifferentiation of the muscles was not affected in either group. These data provide evidence for the susceptibility continuum of CP in this strain. They also indicate that agenesis and hypoplasia of the palatal shelves and primordia of craniofacial skeleton and musculature contribute to CP, the relative involvement of the components depending on the stage of drug administration. In the absence of pronounced cell death, it appears that RA possibly produces its deleterious effects on the precursors of craniofacial primordia, such as the neural crest, by misexpression of developmentally important genes. PMID- 9407596 TI - Birth weight and congenital anomalies following poisonous mushroom intoxication during pregnancy. AB - A series of 22 women who suffered from mushroom poisoning while pregnant have been identified among adults receiving treatment between 1960 and 1993 in a specialist clinic in Budapest, Hungary. In most cases, the poisonings were attributed to Amanita phalloides, verna, and related species. Of these, 20 went to term, and data were collected on gestational age, birth weight, and both major and minor congenital anomalies. Mean birth weight (but not gestational age) was lower than in the control series, suggesting that maternal poisoning may have led to intrauterine growth retardation. Two children were identified with major abnormalities (one of whom had fetal alcohol syndrome related to alcohol abuse by the mother). The prevalences of both major and minor anomalies were similar to the prevalence in the matched control group and to the rate in a more recent control series examined according to the same protocols. However, the statistical power to detect teratogenic effects is limited, especially as only five of the mothers suffered the poisoning episode during the first trimester. PMID- 9407597 TI - Flow cytometric evaluation of the effect of various doses of vindesine sulphate on mouse spermatogenesis. AB - Spermatogenesis, a rapidly proliferating cell system, is highly susceptible to damage by radiotherapy and/or chemotherapy. Vindesine, a semisynthetic vinca alkaloid, was given as a single injection to adult male Swiss albino mice to study its effects on testicular weight and male germ cell turnover pattern using flow cytometry. Testicular weight declined significantly at Day 7 to 14 and from Day 14 to 35 after administration of 1 and 2 mg/kg b wt vindesine, respectively. Flow cytometric evaluation of various testicular cell types after the administration of 2 mg/kg b wt vindesine revealed a significant increase in the relative percentage of spermatogonial cells at Day 21 and 35 posttreatment. In contrast, the relative percentage of primary spermatocytes declined significantly at Day 7 and 14 posttreatment. Similarly, a significant reduction in the relative percentage of round spermatids was observed from Day 7 to 35 posttreatment. The relative percentage of elongated spermatids declined significantly at day 35 post treatment. These changes are reflected in the transformation ratios. While the 4C:2C ratio did not exhibit any significant change below 1 mg/kg vindesine, it declined significantly after 1 mg/kg (Day 14) and 2 mg/kg (Day 7 to 35, except Day 28 posttreatment) vindesine treatment. Treatment of male mice with 2 mg/kg vindesine resulted in a significant decline in 1C:2C ratio from 7 to 35 d post treatment. The 4C:S-phase ratio decreased significantly at Day 7 and 14 posttreatment for all the drug doses above 0.05 mg/kg. A significant reduction in the 1C:4C ratio was observed at day 21 to 35 posttreatment as a result of 2 mg/kg vindesine administration. PMID- 9407598 TI - Leucine sources for 10.5-day rat conceptus in vivo. AB - Protein has been shown to be the principal source of leucine for the Day 8.5 to 10.5 rat conceptus in culture. It could be argued that this finding applies only after adaptation to culture conditions and does not apply in vivo. This possibility was investigated using an isotope-dilution technique after i.v. injection of [3H]leucine into Day 10.5 pregnant rats. Specific radioactivity of free leucine in the conceptus was 8 to 10% of that in maternal plasma. Slow exchange of leucine with the maternal circulation and fetal tissue protein turnover were judged to be inadequate as explanations for the observed isotope dilution. Taken together, our results and those from in vitro studies are consistent with a major contribution of leucine coming from the degradation of protein in vivo, probably involving the visceral yolk sac. Our results suggest that mechanisms of amino acid supply to the conceptus identified using whole embryo culture mirror those in vivo. PMID- 9407599 TI - Estrogen modulation: tiered testing for human hazard evaluation. American Industrial Health Council, Reproductive and Developmental Effects Subcommittee. AB - Recent concerns about the potential of certain chemicals to modulate estrogen regulated processes have led to questions as to how chemicals should be tested for such effects. Therefore, AIHC has developed a comprehensive, resource efficient, and flexible tiered strategy for estrogen modulation (EM) testing. Levels of evaluation include Tier 0, in which exposure, along with alerts based on structure-activity, persistence, bioaccumulation, and other data, are assessed to prioritize chemicals for preliminary testing. In Tier I, short term in vitro, ex vivo, and/or in vivo assays are used to obtain a preliminary indication of EM potential. Among these, an in vivo response assay is considered the most reliable at this time. However, none of these tests are intended for risk assessment, but rather to aid in choosing chemicals for further testing and in guiding the extent of that testing. Tier II is aimed at risk assessment and involves whole animal tests that contain EM-sensitive end points (e.g., two-generation reproduction study). Tier III consists of hypothesis-driven research reserved for situations where targeted research can reduce levels of uncertainty. This tiered approach provides a framework for the strategic and effective application of EM test methods to address specific information needs on a case by case basis. PMID- 9407600 TI - Offspring sex ratio as a potential monitor of reproductive disorders in communities near hazardous chemical sites. PMID- 9407601 TI - Semen analysis of personnel operating military radar equipment. PMID- 9407602 TI - Respiratory rhythm generation in mammals: synaptic and membrane properties. AB - Respiratory rhythm generation depends on a complex interaction between synaptic and membrane properties of functionally defined neurons. To gain a better understanding of how inhibitory and excitatory synaptic inputs lead to the generation of the respiratory rhythm we analyzed the depolarization pattern of respiratory neurons that were recorded in the transverse slice preparation of mice (P8-22) and the in vivo adult cat. Using voltage-calmp recordings from respiratory neurons and specific antagonists for inhibitory synaptic transmission we demonstrate under in vitro conditions, that inspiratory (n = 7) and post inspiratory neurons (n = 13) received concurrent glycinergic and glutamatergic synaptic input during inspiration. A similar conclusion was gained with chloride injections into in vivo respiratory neurons. The inhibitory input was essential not only for generating the characteristic depolarization pattern of respiratory neurons, but also for switching the respiratory rhythm between inspiration and post-inspiration. The generation of the depolarization pattern depends also on intrinsic membrane properties. Negative current injections reveal that excitatory synaptic input was amplified by intrinsic bursting properties in some inspiratory neurons (n = 4) recorded in vitro. Although such properties have not been described under in vivo conditions our findings suggest that with respect to inspiratory, post-inspiratory and late-inspiratory neurons, the principle network organization is similar under both in vitro and in vivo conditions. PMID- 9407603 TI - Regulation of gene expression by hypoxia: a molecular approach. AB - Oxygen is a strict requirement for cell function. The cellular mechanisms by which organisms detect and respond to changes in oxygen tension remain a major unanswered question in pulmonary physiology. Part of the difficulty in addressing this question is due to the limited scope of experiments that can be performed in vivo. In the past few years, several laboratories have begun to make progress in this area, using a variety of cell culture model systems and sophisticated genetic manipulations. Here, we review the current state of knowledge of regulation of gene expression by hypoxia, and describe novel experimental approaches that promise to broaden our understanding of how cells and whole organisms respond to alterations in O2 tension. PMID- 9407604 TI - Molecular aspects of oxygen sensing in physiological adaptation to hypoxia. AB - Oxygen is an essential substrate in aerobic metabolism for most eukaryotic organisms. Thus organisms and cells have developed numerous immediate and long term compensatory mechanisms for dealing with oxygen deprivation. Adaptation to hypoxia at the organismal level includes reflex hyperventilation, polycythemia and angiogenesis, which lead to increased O2 delivery to the tissues. Adaptation at the cellular level involves a shift from oxidative phosphorylation to anaerobic glycolysis, increased glucose metabolism, and expression of hypoxic stress-related proteins. Regulation of many proteins participating in adaptation to hypoxia occurs at the level of gene expression. The most widespread molecular mechanism of hypoxia-dependent regulation is transcriptional induction via the binding of a transcription factor, hypoxia-inducible factor-1 (Hif-1), to the specific sequences on the regulated genes. Long-term induction of many proteins also requires an increase in mRNA stability, which is mediated by the binding of regulatory proteins to specific sequences within the mRNAs. The current theories of coupling between the O2 sensor and mechanisms controlling gene expression are discussed. PMID- 9407605 TI - Intracellular signal pathways controlling respiratory neurons. AB - Medullary respiratory neurons are influenced by a variety of neuromodulators, but there is a lack of information about the specific intracellular signal pathways involved. In this report we describe the modulatory effects of the cyclic adenosine-triphosphate (cAMP)-dependent protein kinase and of protein kinase C pathways on voltage- and ligand-controlled ionic conductances and demonstrate their functional significance in regulating the excitability of medullary respiratory neurons of the vivo cat. Evidence is presented that PKA and PKC pathways are persistently activated. PKA regulates current flow through persistently activated and GABAB receptor-controlled potassium channels as well as GABAA receptor-controlled chloride channels. PKC also depresses persistent potassium currents but it potentiates excitatory and inhibitory synaptic currents. The clinical significance of these intracellular signal pathways is demonstrated in a case of a child suffering from apneustic breathing, who was successfully treated with a 5HT-1A receptor agonist. PMID- 9407606 TI - Developmental modulation of glutamatergic inspiratory drive to hypoglossal motoneurons. AB - Proper function of hypoglossal motoneurons (XII MNs) innervating tongue muscles is critical for respiratory control of the airway. Morphological and electrophysiological properties of XII MNs change during postnatal development, as do modulatory systems. Despite these changes, the system producing respiratory movements must remain fully functional throughout life. Modulatory systems have therefore received considerable attention since coordination of their development with a developing neuromuscular system may be critical for maintenance of continuous, efficient breathing. Developmental modulation of XII inspiratory activity by three transmitter systems is examined. Thyrotropin-releasing hormone (TRH) mediates an increase in MN input resistance (RN) in juvenile but not neonate MNs, and this likely underlies the developmental increase in TRH potentiation of inspiratory activity. Norepinephrine (NE) potentiation of inspiratory activity, which in the neonate is produced in part by an alpha 1 mediated increase in RN, also increases postnatally. Effects of purinergic transmission on XII inspiratory activity remain constant during the first 2 weeks of postnatal development. Adenosine-triphosphate (ATP) produces tonic excitation and inspiratory potentiation that likely result from activation of postsynaptic P2 receptors. A secondary inhibitory effect likely results from hydrolysis of ATP to adenosine and activation of presynaptic A1 adenosine receptors. The functional relevance of these postnatal changes is discussed. PMID- 9407607 TI - Neuromodulation of hypoglossal motoneurons: cellular and developmental mechanisms. AB - Hypoglossal motoneurons (HMs) in the caudal brainstem have a respiratory-related activity pattern and contribute to control of upper airway resistance. In this review, we focus primarily on signalling mechanisms utilized by neurotransmitters to enhance HM excitability. In particular, we consider: (1) the membrane depolarization induced by a number of different putative transmitters [thyrotropin-releasing hormone (TRH), serotonin (5-HT), norepinephrine (NE)]; and (2) the inhibition of a calcium-dependent spike after hyperpolarization (AHP) by 5-HT and its effect on firing behavior. Potential functional consequences on HM behavior of these different neurotransmitter effects is discussed. In addition, we describe postnatal changes in transmitter effects and suggest potential cellular mechanisms to explain those developmental changes. Most of the data discussed are derived from in vitro electrophysiological recordings performed in preparations from neonatal and adult rats. PMID- 9407608 TI - Serotonin receptor mRNA expression in the hypoglossal motor nucleus. AB - Brainstem serotonin (5-HT)-containing cells are remarkable for their widespread axonal projections and having their highest activity during wakefulness and lowest during rapid eye movement sleep. One important site of action of 5-HT is on upper airway motoneurons. However, which of the 14 known 5-HT receptors mediate the effects is uncertain. We used the reverse transcriptase/polymerase chain reaction to detect mRNA for six distinct 5-HT receptors (1A, 1B, 2A, 2C, 3 and 7) in 50 nl micro-punches collected from the hypoglossal (XII) motor nucleus and, for comparison, from the viscerosensory nucleus of the solitary tract (NTS) in adult rats. The relative abundance of the distinct mRNAs was characterized by the minimal number of amplification cycles (25-40) necessary to detect a given mRNA. In the XII nucleus, mRNA for type 1B, 2A and 2C receptors was detectable after 29-31 cycles, detection of type 3 and 7 receptor mRNA required 33-35 cycles; and type 1A receptor mRNA was not detected. In the NTS, detection of mRNA for type 1B, 2C and 7 receptors required 31-33 cycles; type 1A receptor mRNA required 39 cycles; and type 2A receptor mRNA was not detected. The data from the XII nucleus demonstrate that not only the previously recognized type 1B, 2A and 2C receptors, but also type 3 and 7 receptors have the potential to mediate serotonergic effects in XII motoneurons. PMID- 9407609 TI - Modulation of the synaptic drive to respiratory premotor and motor neurons. AB - The characteristics of GABAergic inhibitory modulation of respiratory bulbospinal neuronal activity and short-term potentiation (STP) of phrenic motoneuronal activity were studied. Extracellular unit recording and picoejection techniques in anesthetized dogs showed that both the spontaneous rhythmic and reflexly induced discharge patterns of inspiratory (I) and expiratory (E) premotor neurons were proportionately amplified by the localized application of picomole amounts of bicuculline (Bic), a competitive GABAA antagonist. Intracellular recording and paired-pulse stimulation techniques in anesthetized rats demonstrated an STP of phrenic motor output that appears to be mediated by NMDA receptors and is associated with facilitation of EPSPs and prolonged depolarization of individual phrenic motoneurons. We speculate that both GABAergic gain modulation of premotor neuronal activity and NMDA-mediated STP of phrenic activity may be neural substrates which are involved with the optimization of respiratory and non respiratory behaviors, via adaptive and/or differential control of breathing. PMID- 9407610 TI - Comparative aspects of central CO2 chemoreception. AB - We compare and contrast the putative mechanisms underlying CO2 chemoreceptor function in air breathing vertebrates and terrestrial pulmonate snails. We discuss the role of intracellular pH (pHi) in central respiratory responses to CO2 and describe a variety of patterns of pHi regulation in chemosensory areas. One pattern, in which pHi retains a fixed relationship to the CO2 stimulus over time, seems well suited to chemoreceptor cells. Alphastat regulation of ventilation is apparent in both air breathing vertebrates and terrestrial pulmonate snails. Diethyl pyrocarbonate inhibits respiratory responses to hypercapnia in both groups of animals. The neuronal basis of chemosensitivity is similar, in that putative chemoreceptor cells depolarize during hypercapnic stimulation, but the ionic basis of excitability appears to be a potassium conductance in the vertebrates studied to date and a calcium conductance in the snails. Despite divergent evolutionary histories, chemosensory responses and mechanisms are remarkably similar in air breathing vertebrates and terrestrial pulmonate snails. PMID- 9407612 TI - Life without ventilatory chemosensitivity. AB - In healthy humans ventilatory chemoreception results in exquisite regulation of arterial blood gases during NREM sleep, but during wakefulness other behavioral and arousal-related influences on breathing compete with chemoreceptive respiratory control. This paper examines the extent of chemoreceptive control of breathing within the normal physiological range in awake and sleeping humans and explores the consequences upon breathing of absent chemoreceptive function. Recent studies of subjects with congenital central hypoventilation syndrome (CCHS) demonstrate the extent of behavioral and arousal-related influences on breathing in the absence of arterial blood gas homeostasis. CCHS subjects lack chemoreceptor control of breathing and seriously hypoventilate during NREM sleep, requiring mechanical ventilation. Many CCHS subjects breathe adequately during many waking behaviors associated with arousal, cognitive activity or exercise- presumably reflecting input to the brainstem respiratory complex from the reticular activating system, the forebrain or mechanoreceptor afferents. In most situations, and despite changes in metabolism, the non-chemoreceptive inputs to breathing result in surprisingly well controlled arterial blood gases in CCHS patients. PMID- 9407611 TI - Effect on breathing of surface ventrolateral medullary cooling in awake, anesthetized and asleep goats. AB - In adult and neonatal goats, we chronically implanted thermodes on the ventrolateral (VLM) medullary surface to create reversible neuronal dysfunction and thereby gain insight into the role of superficial VLM neurons in control of breathing in anesthetized, awake and asleep states. Consistent with data of others, cooling caudal area M and rostral area S caused sustained apnea under anesthesia. However, in the awake and NREM sleep states, cooling at this site caused only a modest reduction in breathing, indicating that neurons at this site are not critical for respiratory rhythm in these states. Moreover, data in the awake state over multiple conditions suggest neurons at this site do not integrate all intracranial and carotid chemoreception. The data suggest though that neurons at this site have a facilitatory-like effect on breathing both unrelated and related to intracranial chemoreception. We believe that this facilitation serves a function similar to the facilitation provided by the carotid chemoreceptors and by sources associated with wakefulness. Accordingly, elimination/attenuation of any one of these three influences (caudal M rostral S VLM, wakefulness, carotid chemoreception) results in a slight decrease in breathing, removal of two of the three results in a greater decrease in breathing, and removal of all three results in sustained apnea. PMID- 9407613 TI - Are oxygen dependent K+ channels essential for carotid body chemo-transduction? AB - The mechanism by which the carotid body senses hypoxia and causes an increase in spiking activity on the sinus nerve is not well resolved. Most experimental attention is focused on the glomus cell, a secretory cell which is apposed to the afferent nerve endings and which is the presumed site of oxygen sensing. It is proposed that hypoxia causes glomus cell depolarization by inhibiting an oxygen sensitive K+ current. This leads to depolarization, activation of voltage-gated calcium influx and enhanced secretion of an excitatory transmitter. At present, 4 candidate oxygen-sensitive K+ currents have been identified based on patch-clamp studies of isolated glomus cells. Recent experiments using intact carotid bodies have been undertaken to identify which current is most likely to mediate the hypoxia response. Three of the four currents are sensitive to K+ channel blocking agents (TEA, 4-AP and charybdotoxin), yet all these agents failed to mimic hypoxia, neither stimulating chemoreceptor nerve activity nor enhancing catecholamine secretion. Thus, the fourth current, a leak current which is insensitive to these agents is the most likely candidate for mediating glomus cell depolarization, but the drug-sensitivity of this current is not yet known which precludes a direct test of this speculation. PMID- 9407614 TI - Oxygen-sensing neurons in the caudal hypothalamus and their role in cardiorespiratory control. AB - Work from this laboratory has shown that the caudal hypothalamus modulates the cardiorespiratory responses to hypoxia. The purpose of this review is to describe the modulation of respiratory output by the caudal hypothalamus during hypoxia and how neurons in this area respond to hypoxia. The diaphragmatic activity response to hypoxia was significantly attenuated following microinjection of either cobalt chloride or kynurenic acid into the caudal hypothalamus of rats. In addition, caudal hypothalamic neurons in anesthetized rats and cats responded to hypoxia with an increased firing frequency. This response was maintained in the absence of input from the vagus and carotid sinus nerves in the cat. When recorded extracellularly or by whole-cell patch clamp in vitro, these neurons responded to hypoxia with an increase in firing frequency, membrane potential and inward current. These results suggest that the caudal hypothalamus exerts excitatory influence on respiration during hypoxia, that may originate from the ability of these neurons to sense changes in oxygen levels. PMID- 9407615 TI - Short-term potentiation of carotid sinus nerve inputs to neurons in the nucleus of the solitary tract. AB - Reflex studies have shown that the effects of afferent stimulation can persist beyond the period of stimulation. To determine if some form of 'short-term potentiation' occurs during the initial integration of afferent inputs within the nucleus of the solitary tract (NTS), the synaptic responses of NTS neurons to high frequency carotid sinus nerve (CSN) stimulation were examined in anesthetized rats. In extracellular recording experiments, high frequency CSN stimulation (1-3 sec, 100-300 Hz) increased the number of action potentials evoked by 30 CSN stimuli from 31 +/- 3 to 38 +/- 4 (P < 0.05, n = 11). In this population, evoked discharge was enhanced in six cells, not altered in three cells and reduced in two cells. Spontaneous discharge was increased in the five cells in which it was present (P < 0.05). In intracellular recording experiments, high frequency CSN stimulation increased EPSP amplitude from 5.1 +/- 0.4 to 6.1 +/- 0.4 mV (P < 0.01, n = 21). In this population, amplitude was enhanced in 13 cells, not altered in four cells and reduced in four cells. The enhanced EPSP occurred in the absence of any change in membrane potential in five cells and during a 3-5 mV depolarization in eight cells. In both the intra- and extra cellular experiments, the effects of high frequency stimulation were over within 5 min. The results indicate that brief, intense periods of visceral afferent activation can alter the responses of NTS neurons to subsequent afferent inputs. PMID- 9407616 TI - Chemoreflex and vagal afferent mechanisms enhance breath to breath variability of breathing. AB - In anesthetized rats, vagal afferent activities activate slow central mechanisms which modulate the pattern of breathing over several breaths, giving rise to increased breath to breath variability of respiratory pattern. We hypothesized that variability in breathing pattern would produce variability in blood gases and further enhance breath to breath variability of inspired ventilation. Anesthetized rats were placed in a head-out plethysmograph and spontaneous breathing recorded during inhalation of room air and 100% oxygen. The standard deviations and coefficients of variation of ventilation were similar for both inspired gases, but the shapes of the power spectra of ventilation differed, indicating a relative increase in low-frequency power on room air in those animals exhibiting little low-frequency power on oxygen. Simple indices of variability cannot discriminate these temporal changes in breathing pattern variability. PMID- 9407617 TI - Early ontogeny of rhythm generation and control of breathing. AB - The ability of central networks to produce rhythmic motor behaviours linked to the respiratory function, is a remarkably conserved property of the brainstem reticular formation in vertebrates. Conserved cellular and molecular mechanisms also underlie the early embryonic development of the brainstem, leading to a segmented rhombencephalon in all vertebrates. We have proposed that the neural network that controls breathing after birth, derives from a primordial rhythmic network first active in the segmented hindbrain of the embryo. Observations on transgenic mice support this hypothesis: homozygous inactivation of Krox-20, a gene governing segmentation, leads to a lower-than-normal respiratory frequency (fR), despite fetal maturation of the respiratory network and functional compensatory control after birth. PMID- 9407618 TI - Red nucleus inhibits breathing during hypoxia in neonates. AB - Conscious, anaesthetized and decerebrate young mammals respond to acute hypoxia with an initial increase in breathing followed by a fall to, or to below, pre hypoxic levels--the biphasic ventilatory response. The ventilatory rise is due to sustained hypoxic stimulation of the peripheral chemoreceptors. We present evidence from brain stem and peripheral chemoreflex studies supporting the concept that, in addition to the peripheral chemoreceptors, hypoxia also activates brain stem pathways that inhibit ventilation. A key part of the inhibitory pathway is an area within the red nucleus. Destruction of this area in young decerebrate rabbits abolishes the ventilatory fall during hypoxia, yet has no effect on respiratory control during normoxia and does not affect arterial blood pressure changes in hypoxia. In support of this hypothesis, we report that there are neurones within the red nucleus that increase their discharge in hypoxia. The role of higher brain stem/hypothalamic function in central respiratory control during hypoxia in the fetus and neonate are considered. PMID- 9407619 TI - Developmental plasticity of the hypoxic ventilatory response. AB - This paper will describe recent studies concerning the existence of developmental plasticity in the hypoxic ventilatory control system and the locus of the functional impairment following perinatal sensory suppression. Suppression of peripheral arterial chemoreceptor activity was achieved by exposing rats to hyperoxia (60% O2) for the first month of life; all measurements were conducted 2 5 months after the exposure (perinatal treated rats). Hypoxic (but not hypercapnic) ventilatory responses were severely attenuated in awake perinatal treated rats, but not in rats exposed to hyperoxia as adults, indicating that the persistent effect is unique to development and is not the nonspecific result of O2 toxicity. Impairments of the hypoxic ventilatory response due to changes in pulmonary mechanics, gas exchange or central integration of carotid chemoafferent inputs were all ruled out as primary causal factors. However, a persistent impairment of carotid chemotransduction in perinatal treated rats was apparent. These studies suggest that the hypoxic ventilatory response is susceptible to developmental plasticity, and that a carotid chemoreceptor deficit is the primary cause. These findings may have important clinical implications for patients subjected to excessive O2 therapy during neonatal intensive care. PMID- 9407620 TI - Neonatal conditioning for adult respiratory behavior. AB - Specific challenges and perturbations presented during certain 'sensitive periods' of neonatal life have been shown to modify the structure and function of several physiological systems in such as way as to produce long-term and often permanent changes in the behavioral expression of the system. These same phenomena have been reported in a few studies of respiratory control. We have shown that a conditioning paradigm of intermittent sleep interruption presented during the first 4 weeks of life in rats will increase the number of respiratory pauses (apneas > 1 sec in duration) observed in these animals some 10-12 weeks later. Furthermore, in adult conditioned animals, apneic pauses can be dramatically reduced by a counter-conditioning stimulus (white noise), and abolished completely by barbiturate anesthesia. These observations indicate that the respiratory control system in the brainstem of the neonate contains a degree of developmental plasticity which is experience-dependent and modifiable, and that the adult expression of acquired apneic breathing patterns can be effectively reduced by behavioral methods. PMID- 9407621 TI - Modulation of ventilatory control during exercise. AB - The control of ventilatory responses to mild or moderate dynamic exercise has been the subject of considerable debate for over a century. The prevailing view has been that the ventilatory response to exercise is stereotypical and rather unmalleable. However, paradigms involving novel associations of stimulus inputs have been shown to modulate breathing in short and longer time scales. The scope of this review includes examples of modified ventilatory responses to exercise which have been investigated in terms of neural mechanisms. An attempt to synthesise the available data into a model of neuromodulation is presented. PMID- 9407622 TI - Pulmonary-locomotory interactions in exercising dogs and horses. AB - In exercising quadrupeds, limb movement is often coupled with breathing frequency. This finding has lead some investigators to conclude that locomotory forces, associated with foot plant, abdominal visceral displacements or lumbo sacral flexion, are the primary determinants of airflow generation. Analysis of respiratory muscle electrical activation (EMG) and contraction profiles in chronically instrumented dogs and horses, along with measurements of esophageal pressure (Pes) changes and limb movements, provide evidence that each breath during the exercise hyperpnea is determined by respiratory neuromuscular events. Specifically: (1) Phasic diaphragmatic EMG and tidal shortening are always synchronous with decreases in Pes; (2) decrements in Pes are always associated with inspiratory flow generation; and (3) strict phase coupling between breathing and stride frequency is not obligatory. Thus, although locomotory-associated forces may minimally assist with flow generation, they are not the primary determinants of breathing during exercise. PMID- 9407623 TI - Respiratory-related control of extrinsic tongue muscle activity. AB - The purpose of this brief report is to introduce new evidence showing that the protrudor and retractor muscles of the tongue are co-activated during inspiration in eupnea and hyperpnea in an anesthetized, tracheotomized rat model. We also review previous work on the respiratory related control of the tongue musculature, and briefly consider the clinical significance of this work. The important new findings are that: (1) Both hypoxia and hypercapnia cause parallel increases in drive to the tongue protrudor and retractor muscles (the genioglossus and hyoglossus muscles, respectively); (2) phasic volume feedback inhibits the peak inspiratory activity of both muscles; and (3) the tongue muscles consistently produce a retraction force when the genioglossus and hyoglossus are co-activated, in both animal and human subjects. This latter observation is consistent with previous work showing that the retractor muscles (hyoglossus and styloglossus) develop up to ten times more force than the genioglossus muscle. The possible mechanical consequences of tongue muscle co activation are briefly considered. PMID- 9407624 TI - Do descending influences alternate to produce episodic breathing? AB - This study examines the episodic breathing patterns of three disparate groups of vertebrates. In an in vitro bullfrog brainstem-spinal cord preparation, episodic breathing was replaced by uniformly spaced breaths following transection caudal to the optic chiasma. The same effect was produced in hibernating squirrels by inhalation of mild anesthesia. Preliminary data suggest that a similar conversion is also produced in hibernating squirrels by vagotomy, in conjunction with blockade of central NMDA-type glutamate receptors. In all cases, even though overall breathing frequency increased, due to elimination of periods of apnea, instantaneous breathing frequency slowed. Seals breathe episodically in sleep and when these animals awaken after the start of a breathing episode, breathing also immediately slows. The data presented here are consistent with the suggestion that in all vertebrates, higher centres can modulate the central rhythm generator for breathing, in both a positive and a negative fashion. During episodic breathing, in the species studied here, these modulating influences alternate in a fashion that produces periods of apnea alternating with periods of relatively high frequency ventilation. PMID- 9407625 TI - Establishing the limits and characteristics of normal age-related cognitive decline. AB - BACKGROUND: Research into ageing-related pathology relies not only on exploration of disease aetiology, but also a clear understanding of the normal ageing process. The present study aims to examine the characteristics of elderly subjects who lie on the borderline between normal and pathological ageing. METHOD: Cognitive functioning is examined using computerized neuropsychometric assessment in a population of 833 normal elderly from which a cohort of 397 subjects with sub-clinical cognitive impairment are followed over three years. Subjects receive a standardized neurological examination and ApoE genotypes are established. RESULTS: Analysis of covariance revealed no cross-sectional age differences for syntax comprehension (p = 0.19), articulation (p = 0.46), semantic matching (p = 0.12), reading (p = 0.79), and implicit memory (p = 0.21) while explicit memory, language skills and visuospatial skills were found to deteriorate both in the cross-sectional age comparisons and across time. An overall intellectual ability factor, derived from Principal Components Analysis, was found by regression to decline principally in persons with low education, and a high initial IQ level was observed to provide a protective effect over age 75. Persons with higher levels of education show relative stability over time on language and secondary memory tasks but deteriorate as rapidly as persons with low education on visuospatial tasks. Five separate patterns of sub-clinical cognitive deficit were isolated by cluster analysis. Two groups, with differing clinical profiles (of which only one manifested the ApoE4 allele), were found to have an increased risk of developing senile dementia (OR = 4.4 and 3.9). A third group had a high prevalence of depressive illness, and the remaining two showed very little change. CONCLUSION: Ageing does not affect all cognitive functions uniformly: a high initial education level slowing rate of decline for certain tasks. Separate patterns of cognitive change are observed in early senile dementia, benign change and changes related to depressive illness. Results suggest the need for more stringent selection of normal control groups. PMID- 9407626 TI - Molecular changes during the genesis of human gliomas. AB - Neoplastic transformation in the normal human brain occurs as a result of the accumulation of a series of genetic alterations. These genetic alterations include the loss, gain or amplification of different chromosomes which lead to altered expression of proteins that play important roles in the regulation of cell proliferation. Several common genetic alterations at the chromosomal level (loss of 17p, 13q, 9p, 19, 10, 22q, 18q and amplification of 7 and 12q) have been observed. These alterations lead to changes in the expression of several genes; protein 53 (p53), retinoblastoma (RB), interferon (INF) alpha/beta, cyclic AMP dependent kinase number 2 (CDKN2), mutated in multiple advanced cancers 1 (MMAC1), deleted-in-colon carcinoma (DCC), epidermal growth factor receptor (EGFR), platelet derived growth factor (PDGF), platelet derived growth factor receptor (PDGFR), MDM2, GL1, CDK4 and SAS during the genesis and progression of human gliomas. Recent studies suggest that altered expression of several other genes [MET; MYC; transforming growth factor beta (TGF beta); CD44; vascular endothelial growth factor (VEGF); human neurological-related cell adhesion molecule (hNr-CAM); neuroglial cell adhesion molecule (NCAM L1); p21waf1/Cip1; TRKA; mismatch repair genes (MMR); C4-2; D2-2] and proteins [e.g., cathepsins, tenascin, matrix metalloproteases, tissue inhibitors of metalloproteases, nitric oxide synthase, integrins, interleukin-13 receptor (IL-13R), Connexin43, urokinase-type plasminogen activator receptors (uPARs), extracellular matrix proteins and heat shock proteins] are associated with the genesis of human gliomas. Taken together, these findings point to the accumulation of multiple genetic mutations coupled with extensive changes in gene expression in the etiology of human gliomas. PMID- 9407627 TI - Role of stereotactic biopsy in the diagnosis and management of brain tumors. AB - Stereotactic biopsy has evolved as a powerful and safe tool to provide tissue diagnoses with minimal disruption of normal functioning brain. It plays a significant role in the management of malignant brain tumors, where the benefit of open surgery might not justify the concomitant risks. Stereotactic procedures are closed procedures, and thus direct feedback is not provided to the surgeon during manipulation of brain tissue. This difference from most other neurosurgical procedures necessitates rigor in the preoperative workup, the planning of the procedure, and the conduct of the procedure. The success of the procedure is measured by the ability of the team to make an accurate histopathological diagnosis of the lesion; in experienced hands, the rate of success should exceed 95%. Complications and mortality can be minimized with appropriate attention to detail. PMID- 9407628 TI - Strategies in the surgical management of malignant gliomas. AB - The overall prognosis for patients with malignant gliomas remains poor. The infiltrative nature of the tumor into normal brain and the presence of tumor foci in regions remote from the main tumor burden make cure with current therapies virtually impossible. Management therefore consists of tumor control while maintaining the patient's quality of life. Surgery comprises only one arm of the overall treatment plan. Biopsy allows diagnosis and tumor grading even when tumors are located in eloquent or deep areas of brain. Craniotomy decreases overall tumor burden and provides room for the normal brain, edema and recurrent tumor. Many adjuncts are available to assist in gaining surgical access to tumors with minimal violation of normal functioning brain. Important among these are stereotaxy and surgery within a specially designed magnetic resonance scanner. The strategies for the surgical management of malignant gliomas utilized at our institution will be discussed. PMID- 9407629 TI - Radiotherapy of primary malignant brain tumors. AB - Radiotherapy is usually recommended for patients with a primary malignant brain tumor. The foundation for its use is grounded on the results of randomized trials for malignant gliomas which have demonstrated a relationship between survival and external beam radiation dose. Although similar trials have not been performed for most other primary intracranial tumors, radiation oncologists treat them in a similar fashion, delivering the highest possible dose consistent with acceptable levels of normal tissue damage. In most cases, focal therapy is required, using modern three-dimensional treatment planning and delivery--with whole brain or craniospinal therapy used only for infrequently encountered clinical presentations. With modern planning, the volume of normal tissue subjected to possible radiation damage can be minimized. Radiation effects in normal tissues typically occur months to years after treatment. PMID- 9407630 TI - Radiosurgery for primary malignant brain tumors. AB - Despite the ability of surgery, radiotherapy, and chemotherapy to prolong survival in patients with glioblastoma multiforme (GBM), most patients succumb to their disease, usually as a result of local tumor persistence or recurrence. Stereotactic radiosurgery (SRS) allows a substantial increase in total dose at sites of greatest tumor cell density while sparing most of the normal brain, resulting in significantly improved survival. SRS was designed as a technique to deliver a large single dose of radiation to a small and focal target: two of its hallmarks are the focal distribution of dose and the inverse relationship between dose and volume. Acute complications of SRS are related to edema and are manifested as a worsening of pre-existing symptoms: seizure, aphasia, and motor deficits--these are treatable with steroids and are transient in the majority of cases. The actuarial risk of undergoing reoperation was 33% at 12 months and 48% at 24 months, following SRS. Patterns of failure were similar following brachytherapy or SRS as treatment for recurrent GBM with most patients experiencing marginal failure outside the original treatment volume. Patients with small (< 30 mm diameter), radiographically distinct and focally recurrent GBM should be considered for SRS. Larger lesions (> 30 mm diameter), especially those adjacent to eloquent cortex or critical white matter pathways, must be evaluated with caution. The potential for acute toxicity associated with SRS increases substantially for larger lesions. There is a significant survival advantage using SRS in many patients with gliomas, especially if appropriately used with surgery and other adjuvant therapy. PMID- 9407632 TI - Whole brain radiotherapy in the treatment of metastatic brain tumors. AB - Previous prospective and retrospective trials have failed to demonstrate the best treatment approach for patients with brain metastases. As a result, fractionated whole brain radiotherapy (WBRT) has been the mainstay of treatment for several decades. However, with improved surgical techniques and the advent of radiosurgical procedures to treat single and multiple metastases, the continued value of WBRT is in question. This is particularly true in the treatment of a favorable patient subset where the risks of long-term morbidity need to be addressed. This article reviews the trials of the Radiation Therapy Oncology Group (RTOG) and other select radiotherapy brain metastases trials, and compares their morbidities and outcomes to surgical and radiosurgical techniques. It is unfortunate that the inherent selection bias in most retrospective studies makes comparisons difficult. Therefore, to better understand the roles of WBRT, surgery, and radiosurgery in the treatment of brain metastases, additional randomized studies need to be conducted on homogeneous patient groups. PMID- 9407631 TI - Surgical treatment of metastatic brain tumors. AB - Key elements in the modern surgical treatment of metastatic brain tumors are a firm grasp of criteria for selection of proper surgical candidates and a thorough grounding in the surgical approaches to, and the anatomy of, cerebral metastases. It is important to realize that the presence of multiple or recurrent brain metastases does not automatically contraindicate surgery because in properly selected patients, resection of multiple metastases or reoperation for recurrent metastases can extend survival and enhance the quality of life. Appropriate treatment of metastatic brain tumors frequently requires the judicious use of modalities such as open craniotomy, whole brain radiotherapy, and stereotactic radiosurgery. In order to assure the best outcome of patients with cerebral metastases, it is necessary to have an awareness of how these modalities can best complement one another and to apply them accordingly. PMID- 9407633 TI - Radiosurgery for the treatment of brain metastases. AB - Surgical resection and whole brain radiotherapy (WBRT) have been the mainstays of the treatment of cerebral metastases. This approach results in a median survival of about 10 months. Several recent publications and our own experience suggest that a similar median survival can be achieved with stereotactic radiosurgery using either the Leksell Gamma Knife or the linear accelerator radiosurgical techniques. In addition, radiosurgery can effectively treat metastatic tumors in surgically inaccessible sites, e.g., the brainstem. Radiosurgery can also effectively treat multiple intracranial metastases in widely separated areas of the brain. In fact, we have shown that patients with multiple metastases have similar lengths and qualities of survival as do patients with single metastases treated with stereotactic radiosurgery. The most important predictor of success in radiosurgical treatment of cerebral metastases is the neurological status of the patient, usually expressed as the Karnofsky Performance Status (KPS). The histological type of primary cancer is not an outcome predictor. Even so-called "radioresistant" tumors (e.g., melanoma, renal cell) respond favorable to radiosurgery. A great benefit of radiosurgery is the virtual lack of perioperative complications and the minimal interference with quality of life compared either to surgery or to fractionated whole brain radiotherapy. Long-term complications of radiosurgery are infrequent and primarily relate to failure of local tumor control (10%) and radiation-induced edema or necrosis. The later usually can be controlled with corticosteroids, but occasionally, craniotomy may be required to treat life-threatening mass effects. We believe that radiosurgery is the treatment of choice for most cerebral metastases. Only large lesions (> 3.5-4 cm diameter) and those which require immediate decompression to treat life threatening mass effects require surgical treatment. Radiosurgery also may be used to treat residual disease after surgical resection. We have shown that WBRT does not increase the efficacy of radiosurgery in the treatment of cerebral metastases, and, therefore, we prefer to avoid both the short- and long-term morbidity of that treatment, if possible. PMID- 9407634 TI - Interstitial brachytherapy for malignant brain tumors. AB - For nearly 20 years, interstitial brachytherapy has been used as adjuvant treatment for malignant brain tumors in both prospective clinical trials and as part of standard therapy. Numerous publications analyzing the results of this treatment seem to indicate an improvement in median survival for highly selected patients. Some newly diagnosed glioblastoma multiforme, recurrent malignant glioma, brain metastases and possibly low grade gliomas seem to benefit. While Iodine-125 (I-125) remains the most popular radionuclide for brachytherapy, there is a recent move away from temporary high-activity implants to permanent low activity implants. This review article will concentrate on the results from the University of California, San Francisco, as well as recent series published since 1990. In spite of the increased availability of radiosurgery, interstitial brachytherapy still has a place in the management of these difficult tumors. PMID- 9407635 TI - Chemotherapy of brain tumors. AB - For many years, chemotherapy has been utilized for the treatment of malignant brain tumors with minimal success. This particularly true with chemotherapy in adult malignant gliomas for which no new drug has been approved for use since the initial studies using nitrosoureas and procarbazine in the early 1960s. However, the results of more recent clinical trials research using newer agents appear to show improved outcome in some tumor types. Better understanding of the basic biology of these diverse tumors also have given rise to new treatment strategies, especially to drug development. A large number of new antineoplastic agents are now being tested in Phase I and II trials; they are very promising and soon may lead to new drug approvals. This review will discuss the results of completed trials for newly diagnosed and recurrent glioma in adults, as well as ongoing, new drug studies with these patients. A discussion of the more common pediatric brain tumors and the use of chemotherapy in that age group is also presented. Chemotherapy is often the primary treatment modality used to control tumor growth in newly diagnosed infants and young children, and it is in this setting that chemotherapy is particularly beneficial in reducing morbidity and increasing survival. As part of a multimodality approach that includes surgery and radiotherapy, chemotherapy has a significant role to play in the treatment of both adults and children with brain neoplasms. PMID- 9407636 TI - Moving blood. AB - Our internationally acclaimed journalist Sanguinia has returned safely from her historic assignment. Travelling from Homeric Greece to British Romanticism, she was witness to blood drinking, letting, bathing, and transfusion. In this report, she explores connections between the symbolic and the sadistic; the mythic and the medical--all in an effort to appreciate the layered meanings our culture has given to the movement of blood between our bodies. PMID- 9407637 TI - Collaborative study to assess the suitability of a proposed working reagent for human parvovirus B19 DNA detection in plasma pools by gene amplification techniques. B19 Collaborative Study Group. AB - BACKGROUND AND OBJECTIVES: A collaborative study was done to examine the sensitivity and specificity of assays for the detection of human parvovirus B19 DNA in plasma pools by PCR techniques and to establish a working reagent for B19 DNA testing of plasma pools. MATERIALS AND METHODS: Duplicate samples consisting of a tenfold dilution series of a positive cryosupernatant diluted in B19 DNA negative cryosupernatant were sent to 17 laboratories. RESULTS: The sensitivity of the assays varied: 2 laboratories were able to detect the 10(-7) dilution while 1 laboratory failed to detect B19 DNA in any samples. In addition, 5 laboratories obtained false-positive results. CONCLUSIONS: In general, laboratories using assays optimised for rapid detection of B19 DNA in serum samples did not perform well, indicating that such rapid methods are not adequate for examination of plasma pools. The 10(-6) dilution was detected by approximately half the laboratories and could be used as the working reagent. PMID- 9407638 TI - Clinical and laboratory experience with erythrocyte and platelet preparations from a 0.5CPD Erythro-Sol opti system. AB - BACKGROUND: Red blood cells stored as concentrates or suspensions in additive solutions change rapidly their oxygen affinity mainly due to the loss of 2,3 diphosphoglycerate (2,3-DPG). When collected in CPD with half of the normal concentration of citrate and citric acid (0.5CPD) and stored in a new additive solution (Erythro-Sol), 2,3-DPG is better maintained. No studies of the oxygen affinity of red cells stored under these conditions have been published. In Erythro-Sol, red cells have a satisfactory in vivo recovery for 49 days but the conditions after 28 days, within which time most red cell units are transfused, have not been investigated. Of importance is also to be able to make platelet concentrates (PCs) from 0.5CPD blood. Little data are available concerning the clinical usefulness of platelets prepared from 0.5CPD buffy coats (BCs). METHODS: Blood was collected in 0.5CPD, held at 20 degrees C for 3-4 h, then separated with the bottom-and-top technique into red cells, plasma and BC. In a storage experiment with 6 U the 2,3-DPG and P50 values were determined weekly and a number of in vitro parameters were tested on day 28. In 6 donors the in vivo recovery and survival of red cells were determined using a single-chromium technique. Transfusions of 212 0.5CPD-Erythro-Sol red cell units were given to hematological patients under supervision. PCs derived from pools of 0.5CPD BCs suspended in PAS2 (T-Sol) were transfused to 20 thrombocytopenic patients and compared with CPD-BC-PCs suspended in PASI. Corrected count increments (CCI) were determined. RESULTS: The erythrocyte 2,3-DPG and P50 values were normal or slightly subnormal initially but increased to supernormal levels during the 1 week, and remained at these levels for a further 1-3 weeks; the 2,3-DPG was two thirds of normal after 28 days, the P50 was 3.72 +/- 0.28 kPa after 14 days and 2.84 +/- 0.41 after 28 days (mean +/- SD). The P50 values corresponded closely (r2 = 0.903) to 2,3-DPG. The in vivo recovery of 4-week-stored red cells was 89.6 +/- 5.5% and the T50 was 32.2 +/- 2.0 days. No adverse effects were observed in the transfusions. The CCI values did not differ between test and control groups; in both, 3- to 5-day-stored PCs gave lower CCI than fresh (0-2 days) PCs. Patients with acute myeloid leukemia AML (n = 11) had significantly lower CCI values than patients with myelodysplastic syndrome, myeloma and lymphoma (n = 9; CCI 1 h: p = 0.001; CCI 24 h: p = 0.006). CONCLUSIONS: Red cells stored in Erythro-Sol sustain a normal or slightly lowered oxygen affinity for 2-4 weeks, their viability is excellent, and they are well tolerated in clinical transfusions. Platelets prepared from 0.5CPD-BCs cause CCI, of the same magnitude as CPD-BCs. PMID- 9407639 TI - Demonstration by flow cytometry of the numbers of residual white blood cells and platelets in filtered red blood cell concentrates and plasma preparations. AB - BACKGROUND AND OBJECTIVES: New-generation polyester filters provide significant depletion of white blood cells (WBC) and platelets (PLT) in filtered red blood cell concentrates (FRCC) and in filtered plasma preparations (FP). The aim of this study was to elaborate a sensitive flow cytometric method for monitoring residual WBC and PLT in FRCC and FP. MATERIALS AND METHODS: We determined the number of WBC in 500 microliters FRCC of FP using 50 microliters of a combination of monoclonal antibodies (MAB) against CD45 (FITC labeled) and CD19 (PE labeled). After lysis of red blood cells, we mixed a specific number of reference beads with the remaining WBC. The number of residual WBC related to the acquisition volume was defined by the acquired reference beads. Using this method, the detection limit (DL) was 3 WBC/microliter. Alternative methods used MAB against CD45 (FITC and PerCP labeled) and CD14 (PE labeled) or lymphocyte subsets such as CD3 (FITC labeled) and CD19, CD4, CD8, CD16 and CD56 (PE labeled) in combination with CD45 (PerCP labeled). The DL values were 10 WBC/microliter for the CD45/CD14 staining and 0.1 WBC/microliter for the determination of both CD3+ and CD19+ lymphocytes. For residual PLT in FRCC or FP, we used an FITC-conjugated MAB against CD41, with reference beads to determine the acquisition volume. PLT were demonstrated in a green-fluorescence (FL1) single histogram after gating in the forward light scatter x 90 degrees light scatter signal dot plot. PLT counting was as described for WBC. The DL value was about 2 PLT/microliter. RESULTS: Filtration with Pall WBF-1 filters reduces WBC by 4 log and PLT by 3-4 log, resulting in cell counts which are below the critical limit for causing adverse transfusion reactions. CONCLUSIONS: Flow cytometry techniques provide a reproducible and objective tool for counting residual WBC and PLT in blood preparations compared with the Nageotte hemocytometer. Absolute numbers of leukocyte and lymphocyte subpopulations are obtainable. PMID- 9407640 TI - Evaluation of two different protocols for peripheral blood stem cell collection with the Fresenius AS 104 blood cell separator. AB - OBJECTIVES: Reconstitution of hematopoiesis by means of peripheral blood stem cells is a valid alternative to autologous bone marrow transplantation. The aim of this investigation was to increase the efficiency of collection of circulating blood progenitor cells and to obtain a purer product for transplant. METHODS: We carried out leukapheresis procedures with the Fresenius AS 104 blood cell separator, using two different protocols, the previously used PBSC-LYM and a new mononuclear cell collection program. RESULTS: Both programs were highly effective in collecting mononuclear cells (MNC) and CD34+ cells. Some differences were found, especially regarding MNC yield and efficiencies. There are remarkable differences in the efficiency of collection of CD34+ cells (62.38% with the new program as opposed to 31.69% with the older one). Linear regression analysis showed a negative correlation between blood volume processed and MNC efficiency only for the PBSC-LYM program. Differences were also observed in the degree of inverse correlation existing in both programs between patients' white blood cell precount and MNC collection efficiency. The inverse correlation was stronger for the PBSC-LYM program. Seven patients with solid tumors and hematologic malignancies received high dose chemotherapy and were subsequently transplanted with peripheral blood stem cells collected using the new protocol. All patients obtained a complete and stable engraftment with the reinfusion product collected with one or two leukapheresis procedures. CONCLUSIONS: High efficiencies and yields were observed in the new protocol for MNC and CD34+ cells. These were able to effect rapid and complete bone marrow recovery after myeloablative chemotherapy. PMID- 9407641 TI - Influence of autologous blood transfusion on natural killer and lymphokine activated killer cell activities in cancer surgery. AB - BACKGROUND AND OBJECTIVES: Immunosuppression associated with blood transfusion may influence postoperative infection rates. It may also affect the prognosis of patients treated surgically for colorectal cancer. To control this effect, study protocols have applied autologous blood donation programs, which are thought to be immunologically neutral. However, evidence has emerged that blood donation itself might have suppressive effects on natural killer (NK) cell activities. At present, there are no data available on the effects of autologous blood transfusion on NK or lymphokine-activated killer (LAK) cells. This might be of interest as LAK cells may be active in tumor control. MATERIALS AND METHODS: 26 patients who underwent surgical resection for colorectal cancer, were assigned at random into two groups: (1) autologous blood donation and transfusion, or (2) allogeneic blood transfusion. NK and LAK activities were determined before blood donation, at surgery, and on the 3rd and 8th postoperative day. RESULTS: Blood donation induced a small decrease in NK and LAK activities. The postoperative courses of the two groups differed. In the allogeneic group, NK activity (-50%, p = 0.018) and LAK activity decreased (-60.7%, p = 0.043), whereas in the autologous group the decline in LAK was less pronounced (-33.7%, p = 0.091), and their NK activity even increased (+17.4%, p = 0.315). NK activity was modulated differently in the two study groups (0.0036). Differences in LAK activities were found between the 3rd and 8th day postoperatively (p = 0.354). CONCLUSIONS: In patients receiving autologous blood transfusion, postoperative suppressed NK and LAK activities were modulated. This implies that autologous blood transfusion is not immunologically neutral, but has an intrinsic immunomodulatory potential. PMID- 9407642 TI - Detection of irregular red cell antibodies: more than 3 years of experience with a gel technique and pooled screening cells. AB - BACKGROUND AND OBJECTIVES: The purpose of this study was to evaluate more than 3 years of experience with a gel technique in combination with pooled screening cells for the detection of irregular red cell antibodies. MATERIALS AND METHODS: Conventional serologic methods were used for blood typing, antibody screening and cross-matching until the end of 1992. We introduced the gel technique as a routine assay for antibody detection and identification in 1993. RESULTS: After the tube technique had been abandoned, the number of false-positive antibody screening tests was reduced by 71%, positive antibody screening tests by 33%, enzyme agglutination by 100% and rouleaux reactions and cold-reacting antibodies by more than 50%. There was a 40% increase in first-time detection of clinically relevant antibodies. We saw no increase in delayed haemolytic transfusion reactions. CONCLUSIONS: For the detection of irregular red cell antibodies, pooled screening cells in combination with a gel technique are at least as efficient and safe as a conventional tube technique with unpooled test cells. PMID- 9407643 TI - The serological profile and molecular basis of a new partial D phenotype, DHR. AB - BACKGROUND AND OBJECTIVES: The Rh D antigen comprises a mosaic of at least 30 epitopes expressed on a 30-kD non-glycosylated Rh D polypeptide. The equivalent Rh CeEe polypeptide expressing the Rh C/c and E/e antigens differs in only 36 of the 417 amino acid residues. Partial D individuals have been described who fail to express a number of D epitopes. MATERIALS AND METHODS: Serologic methods were applied with monoclonal anti-D to map epitopes on the red cells of a proposita aberrant D typing. Polymerase chain reaction (PCR) and DNA sequencing were also done. RESULTS: DNA sequence analysis derived by RT-PCR using total RNA isolated from peripheral blood of this person suggests two mechanisms for the genetic basis of this variants: one here gene conversion events result in the replacement of RHD gene exons with the equivalent RHCE exons; the second where point mutation in the RHD gene generates an amino acid substitution in the Rh D protein. CONCLUSIONS: We report here a new partial D, DHR, where a single point mutation (G to A at nucleotide 686) in exon 5 of the RHD gene results in a conservative amino acid substitution (Arg229Lys), in the predicted Rh D protein. This residue is localised on the fourth predicted exofacial loop of the Rh D polypeptide as determined by hydropathy analysis. This substitution results in the lack of epD 1, 2, 12 and 20 (30 epitope model) and indicates the involvement of loop 4, and in particular the requirement of Arg229, in the expression of these epitopes. PMID- 9407644 TI - Quality of fresh-frozen plasma during storage. PMID- 9407645 TI - Absence of seroconversion to hepatitis B and C among blood donors with elevated alanine aminotransferase. PMID- 9407646 TI - Antibodies to hepatitis C virus in Chandigarh blood donors. PMID- 9407647 TI - Concomitant alloanti-Jka in a patient with an IgA autoantibody. PMID- 9407648 TI - A comparison of forgetting in an implicit and explicit memory task. AB - Among possible criteria for distinguishing separate memory systems for implicit and explicit memory is that of substantial differences in either the form or rate of forgetting. Prior literature has claimed both differential forgetting and equivalent forgetting for implicit and explicit tasks. Existing experimental data for word-stem completion and explicit control tasks were reviewed and shown to be inconclusive. Our experiments measure forgetting in comparable implicit and explicit memory tasks of stem completion and stem cued recall. The form and the rate of forgetting are essentially the same for these implicit and explicit tasks. Levels of processing and task conditions differ only in the level of initial learning or availability. Thus, either the implicit and explicit task reflect traces in the same memory system or they reflect traces in different systems that have identical forgetting dynamics. PMID- 9407649 TI - Changes in memory awareness during learning: the acquisition of knowledge by psychology undergraduates. AB - First-year psychology students took multiple-choice examinations following each of 4 lecture courses and 3 laboratory research methods courses. One lecture course was later retested. Students indicated state of memory awareness accompanying each answer: recollective experience (remember), "just know" (know), feeling of familiarity (familiarity), or guess. On the lecture courses, higher performing students differed from other students because they had more remember responses. On research methods, higher performing students differed because they knew more, and in the delayed retest, higher performing students differed because they now knew rather than remembered more. These findings demonstrate a shift from remembering to knowing, dependent upon level attained, type of course, and retention interval, and suggest an underlying shift in knowledge representation from episodic to semantic memory. The authors discuss theoretical and educational implications of the findings. PMID- 9407650 TI - Growth rate of simultaneous masking in cat auditory-nerve fibers: relationship to the growth of basilar-membrane motion and the origin of two-tone suppression. AB - Although many aspects of the mechanisms by which low-frequency sounds exert their powerful masking on responses to high-frequency sounds are well documented and understood, there are few data on the growth of masking for signal frequencies near, but not necessarily at, auditory-nerve-fiber characteristic frequency (CF). Masking of responses to 6- or 8-kHz tones by a continuous 300-Hz band of noise centered at 500 Hz was measured in single auditory-nerve fibers with various CFs. The growth rate of maskings averaged approximately 2 dB/dB, was typically largest for tones about 10% above fiber CF, and decreased at higher and lower frequencies. This pattern of masking versus frequency relative to CF resembles the pattern of compression of the growth of basilar membrane motion versus frequency at a fixed cochlear place. This correspondence supports the hypothesis that the high growth rate of masking by low-frequency sounds is due to the same mechanisms which produce the compression in the growth of basilar membrane motion. PMID- 9407651 TI - Effects of stapedius-muscle contractions on the masking of auditory-nerve responses. AB - There has been little exploration of the mechanisms by which stapedius muscle contractions reduce the masking of responses to high-frequency sounds by low frequency sounds. To fill this gap in knowledge, controlled stapedius contractions were elicited with direct shocks in anesthetized cats, and measurements were made of the effects of these contractions on the masking of single auditory-nerve fibers and on the attenuation of middle-ear transmission. The results show that the stapedius-induced reductions of masking can be much larger than the attenuations of low-frequency sound. With a 300-Hz band of masking noise centered at 500 Hz, and signal tones at 6 or 8 kHz, unmasking effects over 40 dB were observed for sounds 100 dB SPL or less. The data suggest that much larger unmasking might occur. The observed unmasking can be explained completely by a linear stapedius-induced attenuation of sound transmission through the middle ear and a nonlinear growth rate of masking for auditory-nerve fibers. No central effects are required. It is argued that the reduction of the upward spread of masking is probably one of the most important functions of the stapedius muscle. PMID- 9407652 TI - Frequency-dependent enhancement of basilar membrane velocity during olivocochlear bundle stimulation. AB - Basilar membrane (BM) velocity responses were measured in the presence of olivocochlear bundle (OCB) stimulation. Frequency threshold tuning curves (FTCs) were derived from tone-evoked input-output (I/O) functions. Efferent nerve activation produced decreases in velocity amplitude for frequencies around best frequency (BF) at low stimulus levels with little or no effect for stimuli well below the BF. A level-dependent efferent reduction/enhancement of BM velocity was found for certain stimulus frequencies above the BF. Efferent activation either had no effect or caused small reductions in the velocity response produced by low level sound, whereas, at higher stimulus levels, efferent activation increased the velocity response. The derived FTCs, therefore, showed criterion-dependent changes with efferent activation. For low BM criterion velocities, FTCs showed the classic desensitization of the tip region without a shift of BF. Some BM velocity criterion values showed FTCs with an expanded high-frequency response area, also without a shift of BF. The results suggest that the effect of OCB activation changes the gain of the voltage-dependent outer hair cell motility such that BM velocity response near BF is decreased while increasing the response for tones well above BF. PMID- 9407653 TI - Frequency specificity of the human auditory brainstem and middle latency responses to brief tones. I. High-pass noise masking. AB - This study investigated the frequency specificity of the auditory brainstem (ABR) and middle latency (MLR) responses to 500- and 2000-Hz brief tones using high pass noise masking. Stimuli were linear- (2-1-2 cycles) and exact-Blackman- (5 cycles) gated tones presented at 80 dB peak-to-peak equivalent (ppe) SPL. Cochlear contributions to ABR wave V-V' and MLR wave Na-Pa were assessed by the effects of high-pass noise masking on response amplitudes and latencies. The high pass noise results demonstrate that the ABR and the MLR to the 80 dB ppe SPL brief tones show good frequency and place specificity. Changes in ABR or MLR amplitude and latency with high-pass noise masking did not occur as the masker cutoff was decreased from 2 to 3 octaves above the stimulus nominal frequency until it was within one-half octave of this frequency, below which amplitudes rapidly decreased (500- and 2000-Hz tones) and latencies increased (500-Hz tones). No significant differences existed in the frequency specificity of the ABR versus MLR, or in these evoked potentials to exact-Blackman- versus linear gated tones. PMID- 9407654 TI - Frequency specificity of the human auditory brainstem and middle latency responses to brief tones. II. Derived response analyses. AB - This study investigated the frequency specificity of the auditory brainstem (ABR) and middle latency (MLR) responses to 500- and 2000-Hz brief tones using narrow band derived response analyses of the responses recorded in high-pass masking noise [Oates and Stapells, J. Acoust. Soc. Am. 102, 3597-3608 (1997)]. Stimuli were linear- and exact-Blackman-gated tones presented at 80 dB ppe SPI. Cochlear contributions to ABR wave V-V' and MLR wave Na-Pa were assessed by response amplitude profiles as a function of derived band center frequency. The largest amplitudes of waves V and Na-Pa occurred in the 500- and 707-Hz derived bands in response to the exact-Blackman- and linear-gated 500-Hz tones. The peak in the response amplitude profiles for wave V to both 2000-Hz stimuli was seen in the 2000-Hz derived band. For wave Na-Pa, the maxima in the amplitude profiles occurred in the 2000- and 1410-Hz derived bands for the exact-Blackman- and linear-gated tones. Smaller cochlear contributions to the ABR/MLR were also present at 0.5-1 octave above and below the nominal stimulus frequencies. The ABR/MLR to 500- and 2000-Hz 80 dB ppe SPL tones thus shows good frequency specificity, with no significant differences in the frequency specificity of: (1) ABR versus MLR; (2) these evoked potentials to 500-versus 2000-Hz tones; and (3) responses to exact-Blackman- versus linear-gated tones. PMID- 9407655 TI - The effects of click level, click rate, and level of background masking noise on the inferior colliculus potential (ICP) in the normal and carboplatin-treated chinchilla. AB - Carboplatin produces a selective loss of inner hair cells in chinchilla, substantially reducing the amplitude of the compound action potential. A key question that arises from these experiments is: What effect does a reduction in IHC-eighth-nerve fiber input have on the central auditory nervous system? This investigation evaluated the inferior colliculus potential (ICP) in chinchillas treated with carboplatin. The left ear was surgically destroyed and a recording electrode was placed in the left inferior colliculus. Following thirteen days of recovery time, the ICP was recorded in the awake animal. Click level was varied from 10-20 to 80 dB pSPL. Click rate was varied from 10 to 1000 Hz using both conventional averaging and a cross-correlation procedure. Broadband masking noise was varied from 30 to 70 dB SPL with click level held constant at 80 dB pSPL. The dependent variables were the positive peak latency and peak-to-following trough amplitude of the evoked potential. Following baseline studies, the animals were administered carboplatin (50 mg/kg IP) and retested two weeks later. Prior to carboplatin administration, there was an increase in ICP latency and a decrease in ICP amplitude with decreasing stimulus level, increasing rate and increasing noise level. Mean ICP threshold was 30 dB pSPL. Following carboplatin administration, there was little change in threshold or peak latencies. In contrast, the amplitude of the ICP was reduced on average by one-third, although this effect varied considerably across animals. The magnitude of this amplitude decrement was not strongly dependent on click level, click rate, or the level of background noise. PMID- 9407656 TI - Auditory discrimination of material changes in a struck-clamped bar. AB - The principles of theoretical acoustics were applied to approximately reconstruct the sound pressure waveform at the ear as would be generated by an idealized struck-clamped bar. The result is an inharmonic sum of damped sinusoids whose individual acoustic parameters (frequency, intensity, and decay modulus) are, for a fixed geometry and fixed driving force, uniquely determined by the material composition of the bar. In the standard 2IFC procedure, listeners were asked to discriminate changes in material composition based on their perception of the acoustic waveform. Listener strategies for discriminating such changes were estimated by perturbing slightly the individual acoustic parameters from trial to trial and computing correlations with the listener's response [cf. R. A. Lutfi and E. Oh, J. Acoust. Soc. Am. 95, 2963(A) (1994)]. In general, the correlations reveal that listeners fail to make optimal use of the information in the acoustic waveform by tending to give undue weight, for a given material change, to changes in component frequency. In some case, the accompanying reduction in performance efficiency amounted to 80%. PMID- 9407657 TI - Effect of the relative phase of amplitude modulation on the detection of modulation on two carriers. AB - This study examined how effectively information about amplitude modulation (AM) on two carriers is combined, and whether the detection of AM depends on the relative phase of the AM across carriers. Psychometric functions were measured for detecting 5-Hz sinusoidal AM of carriers with frequencies 1100 and 1925 Hz, with a mean level 65 dB SPL for each carrier. The carriers had a duration of 400 ms with 50-ms raised-cosine ramps on either side of this. A single cycle of 5-Hz sinusoidal AM (200 ms in duration) was imposed on the temporal center of the stimulus, with 100-ms steady-state fringes before and after the modulation. The modulators for the two carriers were either in phase or in antiphase. The modulation of each carrier was equally detectable, as determined in a preliminary experiment. A continuous pink noise background was used to mask the outputs of auditory filters tuned between the two carrier frequencies. There was no effect of relative modulator phase. However, performance was consistently better than predicted from the assumption that information about AM from the two carriers is processed independently and combined optimally. The results are discussed in terms of (1) predictions using Dau's "modulation filter bank model" [T. Dau et al., in Psychoacoustics, Speech and Hearing Aids, edited by B. Kollmeier (World Scientific, Singapore, 1996), pp. 45-48], and (2) the fact that relative modulator phase does have an effect on the detection of frequency modulation on two carriers, as found by Furukawa and Moore [J. Acoust. Soc. Am. 100, 2299-2311 (1996)]. PMID- 9407658 TI - Infants' pitch perception: masking by low- and high-frequency noises. AB - The present research employed an operant conditioning procedure typically used with infants to test noise masking of pure tones and tonal complexes in adults and in 7-month-old infants. Adults and infants were presented with either pure tones of 160 and 200 Hz or harmonic tonal complexes with pitches equivalent to 160 and 200 Hz. The tonal complexes did not contain energy at the fundamental frequency. After learning these tasks, subjects in the tonal complex group categorized spectrally varying tonal complexes according to the pitch of the missing fundamental. Stimuli were subsequently presented in combination with either a low- or a high-frequency noise. Both age groups successfully discriminated pure tones when combined with a high-frequency noise but not when combined with a low-frequency noise in the same frequency range as the pure tone. Infants, like adults, successfully categorized harmonic tonal complexes based on the pitch of the missing fundamental when those stimuli were combined with a low frequency noise in the range of the missing fundamental but not when combined with a high-frequency noise which covered the frequency range of the harmonics themselves. These results suggest that infants rely primarily on a central process and not peripherally generated combination tones to hear the pitch of the missing fundamental. PMID- 9407659 TI - Electrode discrimination and speech recognition in postlingually deafened adult cochlear implant subjects. AB - This study investigated the relationship between electrode discrimination and speech recognition in 11 postlingually deafened adult cochlear implant subjects who were implanted with the Nucleus/Cochlear Corporation multichannel device. The discriminability of each electrode included in a subject's clinical map was measured using adaptive and fixed-level discrimination tasks. Considerable variability in electrode discriminability was observed across subjects. Two subjects could discriminate all electrodes, and discrimination performance by the remaining nine subjects varied from near perfect to very poor. In these nine subjects, the results obtained from the discrimination tasks were used to create a map that contained only discriminable electrodes, and subjects' performance on speech recognition tasks using this experimental map was measured. Four different speech recognition tests were administered: a nine-choice closed-set medial vowel recognition task, a 14-choice closed-set medial consonant recognition task, the NU6 Monosyllabic Words Test [T. W. Tillman and T. Carhart, Tech. Rep. No. SAM-TR 66-55, USAF School of Aerospace Medicine, Brooks Air Force Base, Texas (1966)] scored for both words and phonemes correct, and the Central Institute for the Deaf (CID) Everyday Sentences test [H. Davis and S. R. Silverman, Hearing and Deafness (Holt, Rinehart, and Winston, New York, 1978)]. Seven of the nine subjects tested with the experimental map showed significant improvement on at least one speech recognition measure, even though the experimental map contained fewer electrodes than the original map. Three subjects' scores improved significantly on the CID Everyday Sentences test, three subjects' scores improved significantly on the NU6 Monosyllabic Words test, and five subjects' scores improved significantly on the NU6 Monosyllabic Words test scored for phonemes correct. None of the subjects' scores improved significantly on either the vowel or consonant tests. No significant correlation was observed between electrode discrimination ability and speech recognition scores or between electrode discrimination ability and improvement in speech recognition scores when programmed with the experimental map. The results of this study suggest that electrode discrimination tasks may be used to improve speech recognition of some cochlear implant subjects, and that each electrode site does not necessarily provide perceptually distinct information. PMID- 9407660 TI - Information from time-varying vibrotactile stimuli. AB - Experiments have been carried out to investigate the information transfer available via a single vibrator on the fingertip. In a first experiment, for stimuli with durations 80 to 320 ms, discrimination of a one-octave step change in frequency at the halfway point was investigated. Results were similar for three stimulus types--sinewave, monophasic pulse and tetraphasic pulse- suggesting temporal cues are more important than spectral cues in this task. In a second experiment, subjects were required to perceive changes in a sequence of stimulus elements. A presentation rate of 6.25 elements s-1 was found to give better results than a rate of 12.5 elements s-1. In the former case, the potential information transfer per element was estimated to be approximately 1.0 bits, corresponding to an information transfer rate of around 6 bits s-1. Implications for the design of a tactile aid to lipreading are discussed. PMID- 9407661 TI - Estimating parameters for psychometric functions using the four-point sampling method. AB - Although a psychometric function describing a subject's responses to some physical stimuli is of considerable value, characterizing such functions is time consuming and, hence, is not carried out routinely in psychophysical experiments. A principal reason for the lack of efficiency in characterizing a psychometric function is the use of sampling methods that either converge on a single point on the psychometric function, such as the PEST method, or which distribute observations uniformly over a wide range, such as the constant stimuli method. As an alternative, a multimodal four-point sampling method has been proposed [C. F. Lam, J. H. Mills, and J. R. Dubno, J. Acoust. Soc. Am. 99, 3689-3693 (1996)]. A psychometric function is then fitted to the four points (each with several trials) to estimate the threshold and slope parameters of the psychometric function. Adaptive methods, such as the up-down methods [H. Levitt, J. Acoust. Soc. Am. 49, 467-477 (1971)], can be used to provide good initial estimates of the threshold and spread parameters of a psychometric function described by a logistic function. In ongoing studies of age-related changes in auditory masking and discrimination, this new four-point sampling method has been applied to determine psychometric functions for absolute thresholds as a function of duration, thresholds in simultaneous and forward masking, frequency discrimination, and intensity discrimination in both young and aged human subjects. Results indicate that a reduction in data collection time of about 50% with no increase in variance can be achieved. This increase in efficiency applies to simple detection tasks by normal hearing subjects as well as to complex discrimination tasks by older subjects with hearing loss. PMID- 9407662 TI - Speechreading and the structure of the lexicon: computationally modeling the effects of reduced phonetic distinctiveness on lexical uniqueness. AB - A lexical modeling methodology was employed to examine how the distribution of phonemic patterns in the lexicon constrains lexical equivalence under conditions of reduced phonetic distinctiveness experienced by speech-readers. The technique involved (1) selection of a phonemically transcribed machine-readable lexical database, (2) definition of transcription rules based on measures of phonetic similarity, (3) application of the transcription rules to a lexical database and formation of lexical equivalence classes, and (4) computation of three metrics to examine the transcribed lexicon. The metric percent words unique demonstrated that the distribution of words in the language substantially preserves lexical uniqueness across a wide range in the number of potentially available phonemic distinctions. Expected class size demonstrated that if at least 12 phonemic equivalence classes were available, any given word would be highly similar to only a few other words. Percent information extracted (PIE) [D. Carter, Comput. Speech Lang. 2, 1-11 (1987)] provided evidence that high-frequency words tend not to reside in the same lexical equivalence classes as other high-frequency words. The steepness of the functions obtained for each metric shows that small increments in the number of visually perceptible phonemic distinctions can result in substantial changes in lexical uniqueness. PMID- 9407663 TI - Stop-consonant and vowel perception in 3- and 4-year-old children. AB - Recent research on 5- to 11-year-old children's perception of stop consonants and vowels indicates that they can generally identify these sounds with relatively high accuracy from short duration stimulus onsets [Ohde et al., J. Acoust. Soc. Am. 97, 3800-3812 (1995); Ohde et al., J. Acoust. Soc. Am. 100, 3813-3824 (1996)]. The purpose of the current experiments was to determine if younger children, aged 3-4 years, can also recover consonant and vowel features from stimulus onsets. Ten adults, ten 3-year olds, and ten 4-year-olds listened to synthesized syllables composed of combinations of [b d g] and [i u a]. The synthesis parameters included manipulations of the following stimulus variables: formant transition (moving or straight), noise burst (present or absent), and voicing duration (10, 30, or 46 ms). Developmental effects were found for the perception of both stop consonants and vowels. In general, adults identified these sounds at a significantly higher level than children, and perception by 4 year-olds was significantly better than 3-year-olds. A developmental effect of dynamic formant motion was obtained, but it was limited to only the [g] stop consonant. Stimulus duration affected the children's perception of vowels indicating that they may utilize additional auditory information to a much greater extent than adults. The results support the importance of information in stimulus onsets for syllable identification, and developmental changes in sensitivity to these cues for consonant and vowel perception. PMID- 9407664 TI - Perceptual differences in infant cries revealed by modifications of acoustic features. AB - Previous studies of infant cry acoustics and their perceptual significance have remained inconclusive as to the graded nature of cry production and perception and to the exact role and importance of particular acoustic features. In this study, a set of infant cries were digitally analyzed and resynthesized to form natural-sounding cries with varying fundamental frequency (F0), degrees of jitter (period to period variations in F0), and rise time (time for F0 to reach its maximum value). In a perceptual rating task, higher-F0 cries as well as cries with larger amounts of jitter tended to be given more negative ratings than were lower-F0 cries and cries with less jitter, respectively. The perceptual ratings of the rise time manipulations were inconclusive. This study demonstrated a perceptual effect of F0 and jitter independently of other parameters, consistent with current notions of infant cry gradedness. It was also shown that digital signal processing techniques can be fruitfully applied to infant cry research. PMID- 9407665 TI - Do weak syllables count for newborns? AB - Does the newborn's well-known sensitivity to human speech include awareness of the distinction between strong and weak syllables, as has been shown for older infants and adults? The non-nutritive high-amplitude sucking paradigm was used to investigate whether weak syllables play a role in neonate perceptual representation. Two-day-old French infants were tested on their capacity to discriminate phonetically highly varied words containing syllables with various strong vowels versus the weak, reduced vowel schwa in natural, isolated English words. Twenty infants heard lists of weak-strong and lists of strong words (e.g., belief, control, etc. versus nose, dream, etc.) and 20 heard lists of weak-strong and strong-strong words (e.g., belief, control, etc. versus volume, rhubarb, etc.). The results show that weak-strong words were reliably distinguished from strong words, but not from strong-strong words. Taken together, the findings indicate that a weak, reduced vowel is equivalent to a strong, full vowel to the extent that both count as syllabic nuclei. Moreover, this global equivalence in terms of number of syllabic constituents apparently over-rules the more local acoustic difference between strong and weak vowels. The role of syllabic/vocalic information in neonate representation is discussed. PMID- 9407666 TI - Adult and infant perception of two English phones. AB - Previous research has shown that young infants easily discriminate both native and non-native consonant contrasts, but by 10-12 months of age infants perform like adults and easily discriminate only native consonant contrasts. The present study was designed to determine what kind of experience is required to maintain discrimination of native consonants. To address this question, English listeners of three ages (6-8 months, 10-12 months, and adults) were presented with the phonetic difference, [da] vs [t = a]. This distinction occurs in English but is not phonemic: [t = a] occurs when it follows an [s] (as in /sta/). If passive exposure is sufficient to maintain discrimination, all age groups should discriminate [da] vs [t = a]. However, if phonological status plays a role, then older infants and adults should fail. In experiment 1, English adults judged [da] and [t = a] to be equally good instances of the same phonemic category /da/. In an AX procedure in experiment 2, English adults discriminated [da] vs [t = a] better than chance but worse than native phonemic levels. In the Conditioned Head Turn procedure in experiment 3, adults and 6- to 8-month-old infants discriminated [da] vs [t = a], but 10- to 12-month-old infants did not. Taken together, these results are most consistent with the hypothesis that phonological status plays a role in maintaining discrimination of phonetic information. PMID- 9407667 TI - Estimating articulation scores. AB - The ability of listeners to estimate articulation scores for lists of nonsense syllables was evaluated. Normal-hearing subjects were presented with lists of from 50 to 60 nonsense syllables that were degraded with various amounts of noise or filtering and were instructed to estimate consonant-correct scores for each condition. To provide a reference for estimating, subjects were shown the accurate orthographic representation of the syllable on a computer monitor to compare with the auditory presentation. The printed version was displayed either simultaneously with the auditory presentation or 500 ms after the offset of the syllable. Estimates were collected on two occasions to examine test-retest reliability, and actual percent-correct scores were obtained to check the accuracy of the estimates. Most subjects overestimated actual scores when the printed representation was provided simultaneously, but estimates were strikingly similar to actual scores when the printed representation was delayed. The delay appeared to prevent the printed representation from favorably biasing the reception of the syllable. The average of two or three estimates gave highly repeatable results for both visual displays. Crossover frequencies derived from the filtered-speech conditions were within the range reported in the literature. This supports the conclusion that subjects based their estimates on the recognition of speech sounds rather than other percepts associated with the speech-in-noise conditions such as loudness of the noise. The estimation procedure permits the collection of articulation scores in much less time than required by traditional test procedures. PMID- 9407668 TI - Central auditory system plasticity: generalization to novel stimuli following listening training. AB - Behavioral perceptual abilities and neurophysiologic changes observed after listening training can generalize to other stimuli not used in the training paradigm, thereby demonstrating behavioral "transfer of learning" and plasticity in underlying physiologic processes. Nine normal-hearing monolingual English speaking adults were trained to identify a prevoiced labial stop sound (one that is not used phonemically in the English language). After training, the subjects were asked to discriminate and identify a prevoiced alveolar stop. Mismatch negativity cortical evoked responses (MMN) were recorded to both labial and alveolar stimuli before and after training. Behavioral performance and MMNs also were evaluated in an age-matched control group that did not receive training. Listening training improved the experimental group's ability to discriminate and identify an unfamiliar VOT contrast. That enhanced ability transferred from one place of articulation (labial) to another (alveolar). The behavioral training effects were reflected in the MMN, which showed an increase in duration and area when elicited by the training stimuli as well as a decrease in onset latency when elicited by the transfer stimuli. Interestingly, changes in the MMN were largest over the left hemisphere. The results demonstrate that training can generalize to listening situations beyond those used in training sessions, and that the preattentive central neurophysiology underlying perceptual learning are altered through auditory training. PMID- 9407669 TI - Acoustic properties of egg yolk and albumen in the frequency range 20-400 MHz. AB - The acoustic propagation properties of egg yolk and albumen are characterized in the frequency range 20-400 MHz by the bioultrasonic spectroscopy system using an ultrasonic transmission comparison method. Significant differences in the attenuation, velocity, impedance, and density among yolk and thick and outer thin albumen are observed. The acoustic properties of 10% aqueous solutions of ovalbumin and bovine hemoglobin are also measured in order to investigate the contribution of proteins to the acoustic properties of albumen. The differences obtained between thick and outer thin albumen may be mainly due to their macromolecular level structural differences, as their constituents are nearly the same. PMID- 9407671 TI - Frequency tuning of the dolphin's hearing as revealed by auditory brain-stem response with notch-noise masking. AB - Notch-noise masking was used to measure frequency tuning in a dolphin (Tursiops truncatus) in a simultaneous-masking paradigm in conjunction with auditory brain stem evoked potential recording. Measurements were made at probe frequencies of 64, 76, 90, and 108 kHz. The data were analyzed by fitting the rounded-exponent model of the auditory filters to the experimental data. The fitting parameter values corresponded to the filter tuning as follows: QER (center frequency divided by equivalent rectangular bandwidths) of 35 to 36.5 and Q10 dB of 18 to 19 at all tested frequencies. PMID- 9407670 TI - Decorrelation of intravascular echo signals: potentials for blood velocity estimation. AB - When blood particles travel through an intravascular ultrasound imaging plane, the received echo signals decorrelate at a rate that is related to the flow velocity. In this paper, the feasibility of extracting blood velocity from the decorrelation function of radio frequency signals was investigated through theoretical analysis and computer simulation. A computer model based on the impulse response method was developed to generate the ultrasound field of a 30 MHz intravascular transducer. The decorrelation due to the scatterer displacement as well as other nonmotion related decorrelation sources were studied. The computer simulations show that the decorrelation function is linearly related to the lateral displacement. The monotonic relationship between correlation and displacement provides possibilities to estimate flow velocity with decorrelation measurements. Because of the complexity of the beam profile in the near field, assessment of local velocities requires detailed knowledge of the decorrelation at each axial beam position. Sources of signal decorrelation other than the lateral displacement may cause a bias in the decorrelation based velocity measurements. For localized decorrelation estimation, measurement variations in small range windows present a major challenge. An approach based on multiple decorrelation measurements should be adopted in order to reduce the variations. In conclusion, results of this study suggest that it is feasible to measure flow velocity by quantifying the decorrelation of intravascular ultrasound signals from blood. PMID- 9407672 TI - Evolutionary theory and the human family. AB - Emlen's (1995) paper "An evolutionary theory of the family" reviewed existing ideas about the nature of family systems and the reasons why they have evolved in certain animal species. His theorizing led him to propose 15 predictions about how family systems function, based on favorable evidence from various species, mostly birds. While he suggested that these predictions can be applied to the human case, he himself did not attempt to do so. We consider the applicability of Emlen's 15 predictions to the study of human family systems, and find that several aspects of the life history and ecology of Homo sapiens require that they be modified. These considerations include: (1) the importance of intragroup solidarity in the context of intergroup competition: unlike in many other species where dispersal constraints arise from food or breeding site shortages, the primary pressure driving human sociality seems to be competition from other human groups; (2) the complex nature of exchange and reciprocity in human society: reciprocal altruism in particular is integral to human social interaction and leads to a particularly high degree of non-nepotistic helping behavior; and (3) the implications of menopause and the existence of potentially dominant, postreproductive helpers: helpers of this sort have little incentive to disperse or to encourage offspring to disperse, thus greatly increasing family stability. PMID- 9407673 TI - Cloning, inbreeding, and history. PMID- 9407674 TI - Nitric oxide, nitric oxide synthase inhibitors, and anesthetic state. PMID- 9407676 TI - Prediction of bleeding diathesis in patients undergoing cardiopulmonary bypass during cardiac surgery: viscoelastic measures versus routine coagulation test. AB - BACKGROUND: Severe hemorrhagic tendency often complicates cardiopulmonary bypass (CPB) in cardiac surgery. In this study, we compared the effectiveness of thromboelastography (TEG), Sonoclot (SCT), and routine coagulation test (RCT) in the prediction of coagulation defects. METHODS: Forty-three patients undergoing cardiac surgery with CPB were included. Blood for RCT, TEG, and SCT profiles was sampled before systemic heparinization and after protamine administration. Clinically significant bleeding was defined as chest tube drainage in excess of 100 ml/h for 3 consecutive hours or 300 ml/h in 1 h. All coagulation parameters obtained before and after CPB were compared. The sensitivity, specificity, accuracy, false positive, and false negative rate were also calculated and compared. RESULTS: All coagulation tests were within normal range except higher partial thromboplastin time. Variables which were significantly different from those before CPB included platelet count, fibrinogen level, prothrombin time, and thrombin time in RCT, alpha angle and maximum amplitude in TEG, and R2 and peak time in SCT. In the TEG tracing, all variables had high sensitivity, specificity, and accuracy (average 85.4%, 83%, and 83.5% respectively) and low false positive and negative rate (12.5% and 5% respectively). Although SCT had high sensitivity (76.3%) and low false negative rate (6.5%), its specificity and accuracy were all under 50%. CONCLUSIONS: Our data demonstrated that the TEG monitoring is a useful tool for detecting post-CPB bleeding diathesis and can provide much predictive information. RCT and SCT are of limited value because of higher rate of unreliable results. PMID- 9407675 TI - Comparison of neuromuscular action of rocuronium, a new steroidal non depolarizing agent, with vecuronium. AB - BACKGROUND: Rocuronium is a new nondepolarizing muscle relaxant. It features a rapid onset and lack of histamine release: It has an intermediate onset of action as vecuronium. The purpose of this study was to compare the neuromuscular action and condition of intubation after a bolus dose of rocuronium or vecuronium (2 x ED90). We also compared the duration of relaxation after intubation and maintenance doses of each drug. METHODS: Sixty male or female patients, age 18 65, scheduled for elective surgery under general anesthesia were divided randomly into two groups (rocuronium and vecuronium group). All patients were ASA class I II and pre-operative laboratory data were normal. Anesthesia was performed with fentanyl, isoflurane and O2. Rocuronium 0.6 mg/kg (2 x ED90) or vecuronium 0.1 mg/kg (2 x ED90) was given during induction of anesthesia. The response of adductor pollicis was measured with acceleromyography. Neuromuscular block was maintained by bolus injection of rocuronium 0.15 mg/kg or vecuronium 0.025 mg/kg when T1 reached 25% of control. Onset time, duration, recovery indices, intubation condition and T4/T1 ratio to 70% were recorded. Side effects were recorded during the study. RESULTS: The onset time was significantly longer in vecuronium group than that of rocuronium group (102.8 +/- 26.9 s vs. 54.9 +/- 10.9 s, p < 0.05). The clinical durations of action were respectively 44.2 +/- 13.2 min in rocuronium group and 42.5 +/- 9.1 min in vecuronium group (T1 to 25%). The duration of the maintenance were respectively 28.8 +/- 9.5 min in rocuronium group and 26.1 +/- 6.8 min in vecuronium group (T1 to 25%). No adverse effect occurred with either drug. The intubation condition was similar in both groups. CONCLUSIONS: We conclude that rocuronium provides a more rapid onset of action than that of vecuronium. Rocuronium is an intermediate-acting muscle relaxant as vecuronium with good to excellent intubation condition. It may be an useful alternative to vecuronium for rapid tracheal intubation. PMID- 9407678 TI - Intermittent bolus versus patient-controlled epidural morphine for postoperative analgesia. AB - BACKGROUND: Patient-controlled epidural analgesia (PCEA) is a technique that combines the flexibility and convenience of PCA with the intrinsic analgesic efficacy of epidurally administered opioids. The aim of this study is to compare the analgesic and side effects of intermittent bolus injections of epidural morphine with PCEA using morphine during the first 24 h after elective low abdominal surgery. METHODS: Sixty patients were randomly assigned into two groups. Patients in group I (n = 30) received 3 mg epidural morphine at an 8-hour interval after surgery. Patients in group II (n = 30) received 2 mg epidural morphine after surgery and patient-controlled analgesia device was processed to deliver morphine 0.15 mg/h, 0.15 mg/bolus. All patients were observed for pain relief and adverse effects for 24 h. RESULTS: Mean morphine consumption was 9 mg for epidural morphine group and 6.87 +/- 0.27 mg for PCEA group. Although the PCEA group utilized significantly less morphine (p < 0.05), pain and sedation scores were similar in the two groups. Pruritus occurred more frequently in the epidural morphine group (63%) than in the PCEA group (33%). Frequency and severity of nausea and vomiting were similar in the two groups. CONCLUSIONS: PCEA with morphine decreases morphine consumption and with less adverse effects than intermittent bolus of epidural morphine. PCEA with morphine is an acceptable alternative to epidural morphine after low abdominal surgery. PMID- 9407677 TI - Evoked facial nerve EMG and brainstem auditory evoked potential monitoring in cerebellopontine angle tumor resection. AB - BACKGROUND: The preservation of normal nerve function or identification of nerve route is critical in some surgeries of cerebellopontine angle tumors. Over the last 5 years, intraoperative facial nerve electromyogram (EMG) and brainstem auditory evoked potential (BAEP) were applied for evaluation of facial nerve integrity and brainstem function in patients while undergoing resection of cerebellopontine angle (CPA) tumor. This report represents the retrospective analysis of our results. METHODS: The inhalational anesthesia with 1-1.5% isoflurane in pure O2 was used. Muscle relaxation was maintained with continuous infusion of atracurium. The degree of muscle relaxation was aimed at a T4/T1 ratio of train-of-four response more than 20% of the adductus pollicis upon ulnar nerve stimulation at the wrist. In 236 patients suffering from CPA tumor without facial palsy, the EMG of the mentalis muscle ipisilateral to the tumor was obtained through stimulation of the facial nerve. The stimulation was applied with a nerve finder, which delivered an electrical stimulation of a single 2 mamp direct current. The EMG finding was compared with the clinical result. In 198 patients, BAEP was used to monitor the brainstem function during tumor resection. In case of intact hearing the BAEP was taken ipsilateral to the operation side and in case with total hearing loss contralateral BAEP to operation side was used. For BAEP stimulation, 90 db click sound stimulation with frequency of 11.26 Hz was applied to both ears. BAEP signals were obtained and recorded at the mastoid region of either side in reference to the vertex. The EMG and BAEP signals were recorded and saved to an evoked potential monitor. RESULTS: In facial nerve EMG monitoring, there were two false positive and no false negative tests. Except for the two false positive tests, the postoperative clinical results in the other cases were compatible with the intraoperative facial nerve EMG findings. In BAEP monitoring, there were twenty-eight positive tests. CONCLUSIONS: The low incidence of false negative test suggests that facial nerve EMG is valuable in detection of facial nerve function in CPA tumor resection. Intraoperative BAEP abnormality is possibly useful in identifying postoperative brainstem dysfunction. PMID- 9407679 TI - NitroG-L-arginine methyl ester reduces the minimal alveolar concentration of isoflurane in rabbits. AB - BACKGROUND: Recently, some studies suggested that nitric oxide (NO) plays a role as a mediator in the central nociceptive pathways and is possibly involved the mechanisms of anesthesia and wakefulness. Inhibition of the L-arginine-NO pathway in the central nervous system may result in an anesthetic, analgesic, or sedative effect. The aim of the present study was to evaluate the effects of the nitric oxide synthase inhibitor (NOSI), nitroG-L-arginine methyl ester (L-NAME), on the threshold for isoflurane anesthesia in rabbits. METHODS: Sixteen New Zealand rabbits were randomly divided into two groups, with eight rabbits in each group. In the study group, a dose of L-NAME 30 mg/kg was injected i.v. daily as pretreatment on three consecutive days, and the fourth dose of L-NAME was given 30 min before the study began. Normal saline was given to the control group. Data of minimal alveolar concentration (MAC), blood pressure (BP), and heart rate (HR) were collected from both groups. Vital signs, such as EtCO2, O2 saturation, and temperature, were maintained within the normal range. All data were described as mean +/- SEM. Statistical analysis was performed using Student's t-test, where p < 0.05 was considered significant. RESULTS: MAC of isoflurane in the control group was 1.90 +/- 0.12%. MAC of the L-NAME group was 1.70 +/- 0.22%, significantly lower than the control group (p < 0.05). CONCLUSIONS: Our preliminary result shows that the MAC of isoflurane in animals treated with L NAME was lower than that in the control group. It is suggested that inhibition of the nitric oxide pathway may enhance the effect of isoflurane. PMID- 9407680 TI - Anesthetic management in a parturient with progressive systemic sclerosis during cesarean section--a case report. AB - This case report concerns a successful Cesarean section (C/S) delivery in an expectant woman affected with progressive systemic sclerosis (PSS) with clinical manifestations of severe pulmonary hypertension (PH), cor pulmonale, severely restrictive ventilatory impairment, pregnancy-induced hypertension (PIH), and esophageal dysfunction under general anesthesia (GA). This is an extremely rare condition in obstetrics and the victim is usually in a great peril of conception, delivery, surgery and anesthesia because of poor pulmonary and cardiac reserves. We herewith reported our experience in two GAs given uniquely to the same patient who was affected with the disorder and discuss the problem. PMID- 9407681 TI - Anesthetic management in parturients with uterine rupture preoperatively--report of two cases. AB - Uterine rupture is a rare obstetric emergency, and the diagnosis of rupture is not always obvious. High surgical delivery rate today which tends to increase the incidence of the disaster urged us to present this report. Two cases of spontaneous rupture of uterus are described. Case 1 concerns spontaneous rupture of a previously intact uterus; case 2 is a rupture due to placenta percreta. A review relevant to its incidence, risk factors, clinical characteristics, and anesthetic managements is given. PMID- 9407682 TI - Retention of broken central venous catheters in pulmonary artery and inferior vena cava--a case report. AB - Central venous catheterization (CVC) has become an important maneuver both for measuring the central venous pressure and for carrying out long-term intravenous alimentation. Furthermore, a central vein may be needed for the rapid restoration of blood volume in condition of acute hemorrhage with difficulties establishing of a peripheral cannulation. However, complications related to either the venipuncture or the presence of catheters in the central venous system were reported. The case we reported here had two ruptured catheter fragments retained in the great vessels, one in the right subclavian vein and the other one in the femoral vein during CVC. They migrated to the pulmonary artery and inferior vena cava, and were finally removed through open sternotomy with pulmonary arteriotomy and exploration of common iliac vein two weeks later. This was an unusual and possibly fatal complication. PMID- 9407683 TI - Unilateral pulmonary edema during general anesthesia--report of two cases. AB - Unilateral presentation of pulmonary edema, though well known to occur, is an uncommon entity. Previous reviews of this subject have discussed the different etiologies, which include rapid reexpansion of collapsed lung, down lung syndrome (gravitational edema), systemic-to-pulmonary arterial shunts, heart failure, compression or occlusion of pulmonary vasculatures, obstruction of a bronchus and an acute manifestation of neuropulmonary reaction (neurogenic pulmonary edema). Occurrence of this complication during surgery, however, is even rarer. We report 2 cases of unilateral pulmonary edema occurring during general anesthesia for elective surgery. PMID- 9407684 TI - Brachial plexus injury during surgery--report of two cases. AB - Brachial plexus is the most commonly injured peripheral nerve by malposition during operation. We present two cases of transient brachial palsy after surgery under general anesthesia. Symptoms of the first case persisted about 60 min. Electromyography (EMG) and nerve conduction velocity (NCV) revealed no abnormal finding three days later. In the second case, axonal neuropathy was found at left axillary and suprascapular nerves by EMG and NCV three weeks later. Symptoms persisted for three months and had complete remission after conservative treatment. PMID- 9407685 TI - Truncal rigidity as a result of epidural sufentanil--a case report. AB - It is well known that intravenous opioids may cause truncal rigidity. To the best of our knowledge truncal rigidity induced by epidural opioid has never been reported. Recently, we came across an accident of truncal rigidity following epidural sufentanil. The victim was a 65-year-old female who received cholecystectomy, choledochotomy, and cholangiography. For post-operative pain control, an epidural catheter was inserted cephalad [corrected] at L1-2 interspace with a length of 4 cm of the catheter retained in the epidural space. The epidural catheter was secured and tested for correct placement with 3 ml of 2% lidocaine with 1:200,000 epinephrine prior to induction of general anesthesia. No opioid was ever given in the operative course. When the patient was fully awake and complained of wound pain in the recovery room 50 mg of sufentanil in 10 ml normal saline was given via the epidural catheter after a negative evacuation test. About one minute after the epidural shot, she was found to lose consciousness without any slightest warning sign. Truncal rigidity and locked jaw that followed entailed respiratory arrest and rapid deterioration of oxygenation which evidenced a life-threatening airway emergency. It spite of our efforts we could not manage to ventilate her with ordinary means. It was not until the administration of 80 mg of succinylcholine and oral endotracheal intubation could an adequate ventilation be reestablished. She regained spontaneous breathing 15 min after the episode but for safety's sake she remained intubated for 6 h until the dissipation of analgesia. Another test dose was attempted, which reconfirmed that the epidural catheter was in proper position. She stayed in the recovery room for 24 h and returned to ward in satisfactory condition. The incidence disclosed that epidural sufentanil even with a dose as small as 50 micrograms could cause truncal rigidity. Thus when epidural sufentanil is applied for post operative pain control constant vigilance is necessary in order to avoid accident. PMID- 9407686 TI - Bilateral projection of neurones of the C6 segment to S1 and S2 segments of the spinal cord in the cat. AB - Sacral projections of neurones located in the C6 segment of the spinal cord were electrophysiologically investigated in alpha-chloralose anaesthetized cats. The cell bodies were found mainly in lamina VIII and in the ventromedial part of lamina VII of the C6 segment. At the thoracic level their axons descended in lateral funiculi, mostly on both sides and only exceptionally contra- or ipsilaterally. However, bilateral projection to sacral segments was less frequent (25 neurones). It is concluded that axons terminate at different levels on both sides of the spinal cord and only part of them project bilaterally to S1/S2 segments. Conduction velocities calculated for all the axons varied from 38 to 80 m/s and were significantly slower for their distal parts. Therefore it is suggested that descending axons send collaterals at various spinal levels. The presented data indicate the importance of these neurones for interlimb coordination. PMID- 9407687 TI - Gamma activity in the piriform cortex and behavioral thresholds for electrical stimulation in the olfactory bulb. AB - The olfactory bulb was stimulated by trains of electrical pulses in freely moving rats. Evoked responses resembling damped oscillations at the gamma frequency of 30-60 Hz were recorded in the anterior and posterior piriform cortex. Different types of unconditioned sniffing were induced by stimulation of the olfactory bulb. They were similar to those evoked by an odorant (amylacetate) but differed from the behavioral patterns evoked by non-olfactory (auditory) stimulation. Rats were trained to avoid foot-shocks following electrical pulses into the olfactory bulb as conditioned stimulus in a two way shuttle-box paradigm. Threshold electrical intensities for inducing evoked responses in piriform cortex, unconditioned behavior, and learned avoidance were compared. Thresholds for unconditioned and conditioned behavior were significantly higher in comparison with those for evoking gamma discharges in anterior and posterior piriform cortex. The results suggest that fast time-locked synchronization in the gamma range in the piriform cortex induced by synaptic input from theolfactory bulb is not sufficient for inducing corresponding behavior. Thus behavioral detection and probably also olfactory recognition do not seem to be direct consequences of this fast time-locked neural synchronization. Additional neuronal processes that are connected with further elevation of stimulation intensity seem to be necessary for that. PMID- 9407688 TI - The role of pulmonary stretch receptor activation during cough in dogs. AB - The role of pulmonary stretch receptors in the modulation of expiratory muscle activity during cough is controversial. To evaluate their potential influence on expiratory effort during cough, we compared expiratory muscle activity during unobstructed cough to that during obstructed cough in which the trachea was occluded at the end-inspiration and maintained throughout the subsequent expiration. Cough was evoked by mechanical stimulation of the intrathoracic trachea in 9 anesthetized, tracheotomized dogs. Peak triangularis sterni (TS), internal intercostal (IIC) and transversus abdominis (TA) muscle EMG were monitored to assess both rib cage and abdominal muscle activation during expiration. During cough, expiratory activity increased and peak activity shifted from Stage II to Stage I expiration. Peak expiratory muscle activation during unobstructed and occluded coughs were not significantly different: during unobstructed coughs, peak EMG's (mean +/- SE as percent of resting breathing) were TS, 212 +/- 18; IIC, 425 +/- 72; TA, 406 +/- 66; and during obstructed cough: TS, 188 +/- 24; IIC, 365 +/- 44; TA, 387 +/- 77 (n = 9). These data indicate that enhanced vagal stimulation resulting from airway occlusion does not affect expiratory activity during cough. We suggest that during cough, the expiratory muscles are activated in a stereotypical pattern by the neural network generating the cough and this pattern of activation is not affected by phasic vagal input. PMID- 9407690 TI - Changes of intertrial response rate with elapse of time after two-way avoidance trial in rats. AB - The changes of intertrial response (ITR) rate along the intertrial interval duration were examined in 30 rats trained in two-way avoidance. The lowest ITR rate was observed just after a cross-through response terminating a trial. Rats trained with a warning signal of darkness showed over-all higher ITR rates and shorter periods of post-trial reduction of ITR rate than rats trained with a more salient noise warning signal. The period of post-trial low ITR rate lengthened in consecutive sessions indicating the gradual development of a safety state. PMID- 9407689 TI - Involvement of the motor trigeminal nucleus in respiratory phase-switching in the cat. AB - We investigated the hypothesis that the motor trigeminal nucleus, consisting of expiratory motoneurons, might be influential in termination of inspiration. We addressed the issue by comparing the effects on neural respiration of a reversible, unilateral, pharmacologic blockade of the motor trigeminal nucleus (5M), the medial parabrachial nucleus (PB), and of other nearby structures that are neutral for respiration in anesthetized, vagotomized, paralyzed, and ventilated cats. The blockade was achieved by microinjections of 2% xylocaine, laced with Pontamine Sky Blue to identify sites of injections, from the tip of a penetrating microelectrode. Integrated phrenic neurograms were recorded to quantify the time of neural inspiration (TI), expiration (TE), and the peak phrenic amplitude. We found that blockade of the 5M caused a pattern of apneustic respiration, consisting of a selective prolongation of inspiratory phases that were interrupted by short expiratory pauses. In contrast, blockade of the PB resulted in a prolongation of both TI and TE, which corresponds to a mere slowing of respiration. The results confirmed important functions of the rostral pons in ventilatory control but pointed to the 5M rather than PB as a structure underlying the inspiratory off-switch. We conclude that the motor trigeminal nucleus may have a part in the pontine pneumotaxic mechanism. PMID- 9407691 TI - Age-related differences in performance of stereotype arm movements: movement and posture interaction. AB - Postural destabilizations in response to cyclic pull-and-push arm movements were compared in young and elderly subjects, with the goal of determining how age related differences in postural stability influence strategies of cyclic arm movements made at different speeds, against different loads and while standing on support surfaces of different compliances. The results show that elderly subjects performed the experimental task more slowly with a lower mean movement frequency and a smaller amplitude. Despite of this fact, the elderly's upright posture was destabilized by this movement to a greater extent than in young subjects. The older adults exhibited lower damping of the disturbing torques produced by arm movements as evidenced by a higher amplitude of the center of foot pressure excursions. The results document close reciprocal motor and posture interaction and indicate that parameters of the voluntary movement task such as cyclic arm movements might be used as a sensitive measure of postural stability. PMID- 9407692 TI - Digging behaviour and responses to photic and gravitational cues as elements of escape behaviour of bumblebees. AB - We have investigated escape behaviour of workers of two bumblebee species, Bombus terrestris and B. pascuorum, when confined to test tubes plugged with soil and either exposed to sunlight or kept in darkness. In both these situations B. terrestris performed better (i.e. escaped after a shorter time) than B. pascuorum. B. terrestris (but not B. pascuorum) also performed better in darkness than in tubes exposed to sunlight. This implies that in both situations B. terrestris showed higher readiness to dig than B. pascuorum, and that in tubes exposed to sunlight only B. terrestris showed high readiness to display photopositive behaviour as well. B. pascuorum displayed, however, photopositive behaviour in another escape situation: when released in a dark room in front of a vertical array of four sources of white light. In that situation, B. pascuorum also displayed the tendency to fly upwards, based most probably on responses to gravitational cues. PMID- 9407693 TI - Naloxone impairs spatial performance in rats. AB - Naloxone-injected (1.0 mg/kg) and saline-injected control rats were subjected to a two-trial test of object localization memory. In trial I rats were allowed to explore for 5 min an enclosed T-maze with an object (plastic bottle) placed in one maze arm. Then, the object was removed and after a 20-min retention interval rats were faced with two empty arms of the same maze (trial II). Control rats showed good retention of the place occupied by the object, displaying a significant preference (74%, P < 0.032) for the arm which previously contained the object. Naloxone treated rats responded at chance levels (53%). An accurate performance in this task is normally based on information provided by spatial cues outside the maze (cognitive map), so that a random performance of Naloxone treated rats could supposedly be related to some disorders in the internal representation of the environment. PMID- 9407694 TI - A simple, non-invasive method for the measurement of reserpine-induced tremor in rats. AB - A new, non-invasive method for measuring reserpine-induced tremor in rodents is described here. The test procedure is based on the piezo-electric principle and was evaluated using the tremorogenic compound reserpine and the stereotypies inducing drug apomorphine. Whereas for reserpine an orderly and dose-related increase in activity was observed, no such effect was detected with apomorphine. In order to further evaluate the test procedure, studies on the antagonism of reserpine-induced tremor were also performed. Results from these studies indicated that the DA-agonist lisuride, but not the S2-antagonist ritanserin, were able to antagonize the reserpine-induced tremor in a dose-related manner. PMID- 9407695 TI - Molecular analysis of prion protein (PrP) and glial fibrillary acidic protein (GFAP) transcripts in experimental Creutzfeldt-Jakob disease in mice. AB - Prion protein (PrPsc) which accumulates in the brains affected with subacute spongiform encephalopathies (SSE) is altered isoform of normal, cellular isoform (PrPc), and PrP deposition is accompanied with spongiosis and astrogliosis. To find the amounts of PrP and GFAP transcripts during progression of experimental Creutzfeldt-Jakob disease we performed comparative RT-PCR on the terminally sick mice brains, 22 weeks following inoculation with Fujisaki strain of CJD agent, and on control brains. The intensity of bands for PrP-mRNA and control beta-actin were similar for infected and uninfected brains, while amounts of transcripts for GFAP increased as for cytokines released by glial cells-TNF-alpha and IL-1 alpha. This study supports thesis that PrPc to PrPsc conversion is post-translational process not related to PrP overproduction. Increased amounts of GFAP-mRNA during the course of the disease correlated with astrocytosis estimated by immunohistochemistry with anti-GFAP antibody. PMID- 9407697 TI - Distribution of compound potentials recorded from the surface of isolated stellate ganglia following stimulation of postganglionic branches, in rats and cats. AB - The experiments were performed on 9 cat and 18 rat isolated stellate ganglia. Rats and cats were anesthetized with alpha-glucochloralose or urethane, respectively. The ganglia, isolated with their branches, were transferred to a recording chamber and constantly superfused with artificial extracellular fluid bubbled with 95% O2 and 5% CO2. Branches of the ganglion were one by one placed in suction electrodes and stimulated. Antidromic evoked potentials were systematically recorded from numerous points on the ganglion surface. The area under the curve of the negative wave of each recorded potential was considered proportional to the number of neurons located in the vicinity of the recording electrode, projecting to the stimulated nerve. We have found that: (1) cardiac sympathetic neurons are located in the lower, caudal half of the ganglia; (2) vertebral sympathetic neurons occupy the cranial, upper half of the ganglia; (3) neurons with axons in the ansae are positioned near the point of exit of the respective ansa from the ganglion; (4) localization of neurons projecting to the same branches is very similar on both sides--right and left; (5) this localization is also similar in rats compared to cats. PMID- 9407696 TI - The effect of corticotropin-releasing factor (CRF) on the gonadotropin hormone releasing hormone (GnRH) hypothalamic neuronal system during preovulatory period in the ewe. AB - Effects of infusions of corticotropin releasing factor (CRF) into the 3rd ventricle of the brain of ewes during the proestrus on the immunoreactive (ir) gonadotropin hormone releasing hormone (GnRH) neuronal system, pituitary luteinizing hormone(LH) producing cells and LH concentrations in the blood plasma were studied. None of the CRF-treated sheep displayed the estrous activity nor ovulated on the day of estrus (17th day of the cycle), and two days later when they were slaughtered. The GnRH center of CRF treated ewes situated in the preoptico-septal area was well organized, but irGnRH stores in the median eminence were low in comparison to the controls (sheep from the late follicular phase of the estrous cycle). The feature and the number of LH-cells in CRF treated ewes were typical for the preovulatory phase of the cycle but the plasma concentrations of LH did not exceed basal levels. These results suggest that CRF induced decrease of irGnRH stores in the nerve terminals of the ME can be responsible for the blockade of the preovulatory surge of GnRH/LH in the sheep. PMID- 9407698 TI - Spike-wave discharges and sleep spindles in rats. AB - Sleep spindles and spike-wave discharges are thought to originate from the same thalamic pacemaker. In the present work it is investigated whether sleep spindles and spike-wave discharges are also sensitive for the same drugs. Adult male WAG/Rij rats were chronically implanted with frontal and occipital EEG electrode pairs. Rats were intraperitoneally injected with clonidine (0.00625 mg/kg), phenobarbital (20 mg/kg), flunitrazepam (0.188 mg/kg). Frontal and occipital sleep spindles and mainly frontal spike-wave discharges were seen in the electroencephalogram. Phenobarbital and flunitrazepam reduced the number of spike wave discharges and enhanced frontal sleep spindles, while clonidine facilitated spike-wave discharges and reduced frontal sleep spindles. The results of these three drugs indicate a reciprocal relationship between the number of frontal sleep spindles and the number of spike-wave discharges. Only clonidine facilitated occipital sleep spindles without an effect on spike-wave discharges. It can be concluded that frontal and occipital sleep spindles have a different pharmacological profile. Furthermore, the inverse relationship between frontal sleep spindles and spike-wave discharges may suggest that sleep spindles and spike-wave discharges are controlled by a single controlling system. However, in order to explain the clonidine data on occipital sleep spindles another factor must be incorporated in properties of the mechanism(s) involved in EEG oscillations. PMID- 9407699 TI - The effect of naloxone on object exploration, object recognition and other types of spontaneous behavior. AB - Object exploration was examined in naloxone injected (1 mg/kg or 4 mg/kg) and saline control rats. Naloxone rats explored an object for a shorter time than did controls, thus indicating a lower investigatory motivation. This effect was dose dependent. Higher drug dose (4 mg/kg) decreased the number of contacts with an object. Both doses increased the mean duration of contacts with an object. The naloxone groups showed intact recognition of a familiar object paired with a new one in two sessions--4 h and 24 h after the injections. The higher drug dose depressed the locomotor activity and wall leaning. Grooming was not influenced by naloxone. The normal daily fluctuations in the level of grooming and locomotion were distorted following the injection of the higher dose of naloxone. The lower dose (1 mg/kg) did not affect the rats' performance in some tests. The results could be viewed as a naloxone-related depression of the behavior containing motor elements like locomotion, wall leaning and object approaching. The prolonged contact time with an object could be the result of a lowered flexibility of movement. However, the decrease of rewarding value of exploration could not be ruled out. Possibly, naloxone exerts several different interacting behavioral effects. PMID- 9407700 TI - The effect of past and present group size on responses to prey in the ant Formica polyctena Forst. AB - We investigated the responses to insect prey (dead houseflies) in 24 "derivative groups" of workers of the ant Formica polyctena created by taking sets of 25 workers out of nine larger "initial groups" kept in laboratory without queens and brood during the preceding five months. In the derivative groups the ants ceased to retrieve flies to their nests after a period ranging from few days to several weeks. The duration of that period did not depend on the present size of the derivative group (decreasing as a result of worker mortality), but was positively correlated with the estimated size of the initial group of the tested ants. The readiness to display venom spraying was higher in smaller derivative groups. These data demonstrate that responses of F. polyctena to insect prey are strongly influenced both by the present and the past size of their group. PMID- 9407702 TI - Laboratory observations of nuptial flights of the ant Polyrhachis laboriosa. AB - Ethology of Polyrhachis laboriosa, an ant species from equatorial Africa, is little known. No field observation of a nuptial flight of these ants was ever made. We describe two nuptial flights observed in a laboratory colony of P. laboriosa at a 3 days interval. They both occurred in the morning while the nest was kept in near darkness (less than 2 lux of daylight). Flying activity of the alates was suppressed within 1 h by their exposure to daylight of about 140 lux, and within several minutes by their exposure to a lamp emitting white light of 3,000 lux and acting as a source of heat. On the day following the first flight the alates and the workers showed exceptionally high level of mutual grooming. The alates, in particular the males, were transported by workers to the brood chambers whenever they strayed outside and after the nuptial flights. PMID- 9407701 TI - Zopiclone versus diazepam effects on EEG power maps in healthy volunteers. AB - EEG effects of zopiclone (7.5 mg), a cyclopyrrolone derivative with hypnotic action, were compared with effects of diazepam (10 mg). Multichannel EEG recordings, double-blind crossover trials with placebo, and oral single doses were used in healthy volunteers. Vigilance-controlled EEG before and after zopiclone (and placebo), and before and after diazepam (and placebo) were analyzed into FFT power spectra. Effects were assessed as placebo-referred pre post-medication power differences in four frequency bands. Overall statistics showed significant (P < 0.007) global differences between medication effects in the delta frequency band (0.5-3.5 Hz). After zopiclone, fronto-central delta increased bilaterally, whereas after diazepam delta decreased over centro parietal to right temporo-occipital regions. These spatially different brain electric effects show that different neuronal populations must have become active in response to zopiclone and diazepam. PMID- 9407703 TI - Thalamic and amygdaloid connections of the auditory association cortex of the superior temporal gyrus in rhesus monkey (Macaca mulatta). AB - Thalamic and amygdaloid connections of three association auditory areas (AA1, AA2, AA3) of the superior temporal gyrus (STG) were investigated. In order to define the projections of the particular areas, injections of fluorescent tracers were made in three monkeys. Distribution of labeling indicates that area AA1 differs from areas AA2 and AA3 in patterns of both thalamo-cortical and amygdalo cortical connections. Area AA1 receives its predominant inputs from the ventral and dorsal nuclei of the medical geniculate body (MGB). The amygdaloid projection to the area AA1 originates from the basal nuclei, whereas input from the lateral nucleus was not found. The characteristic thalamic projections to areas AA2 and AA3 originate from the dorsal MGB nucleus and the polymodal nuclei of the posterior thalamus. The density of projections from the dorsal nucleus gradually decreases from area AA1 to area AA3 while projections from the Plm, Sg and Lim nuclei increase in the same direction. Areas AA2 and AA3 are the source of strong connections with the lateral nucleus of amygdala, which density increases progressively when injections shift from area AA2 to AA3. The basal and accessory basal nuclei are the source of a less significant amygdalofugal projections to both cortical areas. Thus, our experimental data indicate that influence of the polymodal thalamic nuclei increases substantially in the direction of the higher order association areas. The strong relation of the same cortical areas with the lateral amygdaloid nucleus might suggest that areas AA2 and AA3, in addition to auditory input are the site of transfer of complex sensory information to the amygdala. PMID- 9407704 TI - Lamina IV-VI neurones of the second sacral segment projecting to the sixth cervical segment of the cat's spinal cord. AB - Cervical projections of neurones located in the second sacral segment of the spinal cord were electrophysiologically investigated in 12 adult cats under alpha chloralose anaesthesia. Extracellular or intracellular antidromic potentials from 41 neurons following stimulation of their axons in the spinal grey matter in the C6 segment brought evidence of sacro-cervical connections. Cell bodies of the investigated neurones were found mainly in the medial part of laminae IV, V and VI. Axons of these neurones ran in lateral funiculi, mostly in their dorsal parts (dlf). Most of the axons (26) ascended bilaterally. Fibres of 15 cells ran unilaterally--on the ipsilateral (13) or the contralateral side (2). Conduction velocities of axons measured between S2 and C6 segments were in the range from 38.4 to 69.1 m/s. It is suggested that these neurones may play an important role in movement coordination between hindlimbs and forelimbs. PMID- 9407705 TI - Electrophysiological investigations of the mild spinal cord injury in rats. AB - Somatosensory evoked potentials (SEPs) waveform recorded from the normal and injured spinal cord in rats were analyzed. A mild compression spinal cord injury was performed in two experimental models. The first model was used to assess the disappearance and the second one the return of the electrophysiological spinal cord function after injury. In the first experimental model typical changes in waveforms morphology from normal to isoelectric line during spinal cord compression were investigated. In the second model an isoelectric line was achieved by more severe but a shorter time of compression and the typical changes in SEPs morphology after decompression were described. Our evidence suggests that the spinal cord function disappears gradually during mild compression and can return early after decompression. The amplitude after decompression first improved and then gradually deteriorated which is probably caused by secondary insult. Experimental models are both effective and simple and may be used to evaluate the effect of treatment of spinal cord injuries. PMID- 9407706 TI - Photochemically-induced vascular damage in brain cortex. Transmission and scanning electron microscopy study. AB - Morphological changes of microvessels of cerebral cortex were evaluated in a model of cerebral infarction initiated by a photochemical reaction. Rats were treated with intravenous injection of rose Bengal and irradiated from a halogen lamp source through an intact cranium to precipitate microvascular damage. Investigations in transmission and scanning electron microscopy revealed platelet aggregation on endothelial cells preceded by its early ultrastructural damage. Other typical microscopic features of brain ischaemic injury were present suggesting that the present method may be used as a model for investigating ischaemic brain damage. Since the photochemical activation of the rose Bengal dye results in formation of reactive oxygen species this model may be particularly useful to elucidate the role of free radical-mediated endothelial damage in the formation of microthrombi and blood-brain-barrier integrity. PMID- 9407707 TI - Action of serotonin on the laryngeal airway in anaesthetized cats. AB - The pulmonary chemoreflex induced by an intravenous injection of serotonin (5 hydroxytryptamine) in cats consists of prompt apnoea, bradycardia and hypotension, followed by rapid, shallow breathing. The present study had two purposes (1) to compare the effect of 5HT on ventilation and laryngeal resistance in cats and (2) to assess the role of laryngeal afferents in these responses. The effects of an intravenous injection of serotonin at a dose of 0.05 mg per kg of body weight were studied in eighteen anaesthetized cats, breathing spontaneously via a tracheal cannula. In eleven cats the larynx was isolated in situ to measure laryngeal resistance. In post-serotonin apnoea, the expiratory laryngeal resistance rose four-fold. This coincided with the increased afferent activity of the superior laryngeal nerve. In the initial phase of resumed shallow breathing, the increase in the expiratory laryngeal resistance was coupled with reduced tidal volume. Bilateral section of the superior laryngeal nerve failed to affect laryngeal constriction and the ventilatory response to serotonin. Thus laryngeal afferents running within the superior laryngeal nerve are not essential for the respiratory phenomena induced by serotonin. PMID- 9407708 TI - Controversies about iron in parkinsonian and control substantia nigra. AB - According to the oxidative stress theory iron may play an important role in the pathogenesis of neurodegenerative diseases, as e.g. Parkinson's disease (PD). This review presents the results of studies, obtained by various methods, of iron in substantia nigra (SN)--a cerebral structure which degenerates in PD--and shows controversies concerning the amount of iron, its redox state, and the iron binding compounds. Taking into account all published experimental results, the increase in the concentration of iron in parkinsonian SN vs. control may be estimated as (3 +/- 5)%. The presence of large amounts of divalent iron in post mortem SN can be unequivocally negated. It is, however, still possible that iron is involved in the pathogenesis of PD, as even minor changes in the amount and form of iron may initiate processes leading to cells death. PMID- 9407709 TI - CGP 42112A abolishes facilitation of recognition caused by angiotensin II and angiotensin II(3-7) in rats. AB - The role of the angiotensin AT2 receptors in some behavioural effects of angiotensin II (Ang II) and its 3-7 fragment [Ang II(3-7)], using their selective antagonist CGP 42112A, was assessed. Ang II and Ang II(3-7), given intracerebroventricularly (icv) at the dose of 1 nmole each, substantially improved object recognition memory and enhanced apomorphine (1 mg/kg) stereotypy. Pre-treatment of rats with CGP 42112A (2 micrograms), per se ineffective in all tests, abolished activity of both peptides. None of the treatments significantly changed behaviour of rats in open field. The results point to the considerable involvement of the AT2 angiotensin receptors in the improvement of recognition memory caused by Ang II and Ang II(3-7). PMID- 9407710 TI - Direction discrimination learning in normal and visually deprived cats and the effects of lateral suprasylvian lesions. AB - We used 5 binocularly deprived cats (BD cats), 4 control cats reared also in the laboratory (C cats) and 4 cats reared in a normal environment (N cats). The cats were trained to discriminate an upward or downward-moving light spot versus a stationary spot (detection task) and then an upward versus a downward spot (direction task). The N and C cats learned slowly. The learning was slower than in previously studied discriminations of stationary stimuli. However, all N and C cats mastered the detection task and except one C cat the direction task. In contrast, 4 BD cats failed in the detection task and all in the direction task. This result is consistent with single-cell recording data showing impairment of direction analysis in the visual system in BD cats. After completing the training the upper part of the middle suprasylvian sulcus was removed unilaterally in 7 cats and bilaterally in 6 cats. Surprisingly, the unilateral lesions were more effective: the clear-cut retention deficits were found in 5 cats lesioned unilaterally, whereas only in one cat lesioned bilaterally. PMID- 9407711 TI - Interneuronal cortical connections and intertrial responses in appetitive instrumental learning. AB - The organization of interneuronal cortical connections in intertrial periods was studied in 4 cats trained to perform the delayed appetitive instrumental response to a visual conditioned stimulus (CS). Crosscorrelationl analysis revealed changes in intra- and intercortical neuronal networks of the visual and motor cortical projection areas. Depending on the form of behavior in the intertrial period, i.e., the presence or absence of the acquired instrumental response, the functional connections of either informational (time delays of less than 30 ms) or motivational (time delays in the range of 60-100 ms) character dominated between the neurons of the motor and visual cortical areas. PMID- 9407712 TI - Measuring of the postcopulatory departure in male rats. AB - Spontaneous departure from the female was observed in male rats during testing of instrumental sexual responses. Male rats were tested in an apparatus consisting of two compartments connected by a runway composed of four parallel corridors. The final corridor of the runway and the goal compartment as well as the goal compartment and the start compartment were connected by one-way doors enabling the male to run only in one direction. An incentive female was tethered in the goal compartment. After contact with the female, lasting up to the end of a mount bout, or after exploration of the goal compartment, if the copulatory behaviour was not displayed, the male spontaneously passed to the start compartment and a new run started thereafter. The male thus performed the runs through the apparatus without any intervention from the experimenter. PMID- 9407713 TI - Synaptosomal sodium pump activity depends on microfilament cytoskeleton integrity. PMID- 9407714 TI - Phenotypes of individuals with a beta thal classical allele associated either with a beta thal silent allele or with alpha globin gene triplication. AB - BACKGROUND AND OBJECTIVE: beta thalassemia intermedia has its origins in compound heterozygosity for many different beta thal defects or in an interaction of a beta thal defect with altered alpha cluster. Two specific genetic associations (beta thal/beta(+) -101 C-->T and beta thal + alpha alpha alpha or alpha alpha alpha alpha) have been described in recent years as being determining a phenotype similar to that of simple beta thal heterozygote or, alternatively, a clinical picture of thalassemia intermedia. METHODS: A detailed study on this subject was carried out on 55 patients divided into 2 groups. Group I consisted of 20 patients, 17 of whom (Group Ia) had a beta thal/beta(+) -101 C-->T genotype and 3 (Group Ib) had a beta thal/beta IVS II-844 C-->G genotype. Group II consisted of 35 patients with beta thal association + alpha alpha alpha or alpha alpha alpha alpha. The methods of study have already been described in a previous issue. RESULTS: Thirty percent of group Ia and 25% of group II were virtually asymptomatic, while the others presented the thalassemia intermedia phenotype. This second phenotype is generally milder in patients of group I and even less so in those of group II. In the former there is a higher level of HbF; in the second there is more marked alpha/beta + gamma globin synthesis imbalance. The severity of the phenotype has no connection with that of the beta thal defect. The patients of group Ib all presented thalassemia intermedia. INTERPRETATION AND CONCLUSIONS: The definite clinical pictures of groups I and II are quite common in the Italian population and should therefore be well understood, especially for proper application of preventive measures against thalassemia major. PMID- 9407715 TI - A micro colorimetric assay using cryopreserved monocytes to evaluate antibody mediated red cell-monocyte interaction. AB - BACKGROUND AND OBJECTIVE: A number of in vitro assays based on the interaction of red cells with monocytes have been used to determine the clinical significance of red cell antibodies. When used in our laboratory, one of these assays (the monocyte-macrophage phagocytosis assay-MMPA), was very time consuming and showed great variability. METHODS: We set up a monocyte phagocytosis colorimetric assay (MPCA), using standard microtiter plate wells coated at 37 degrees C for 1 hour with monocytes from healthy donors. After washing the wells to remove non adherent monocytes, test red cells are added to the wells. Sensitized red cells bind to the monocytes, which are lysed after incubation to measure red cell phagocytosis. This is done by hemoglobin detection in the lysate through reaction with o-phenylenediamine and absorbance evaluation with a colorimeter. The results are expressed as the phagocytosis index (PI), which is calculated with the following formula: PI = [1-(A450 unsensitized red cells/A450 sensitized red cells)] x 100. In this study we determined: the source of MPCA variability; the precision of MPCA results; the correlation between MMPA and MPCA results; the MPCA reference values and the MPCA and MMPA execution times. RESULTS: MPCA variability depended largely on the monocyte source. The smallest variation coefficient of the results of replicate assays (19-21%) was found using pooled, cryopreserved monocytes. When performed with a pool of cryopreserved monocytes from 10 subjects, the SD of PI values obtained in replicate assays showed little variation (11-13) over the range of anti-D concentrations tested (from 18.75 to 300 ng/mL). A linear correlation coefficient r of 0.96 was obtained when MPCA and MMPA were performed in parallel, and the 95th centile of PI reference values determined with red cells of 40 non-transfused surgical patients free of irregular red cell antibodies was 7. MPCA execution time was 56% of that needed to perform MMPA. INTERPRETATION AND CONCLUSIONS: These studies show that MPCA is an easy and reproducible assay which allows objective and automated evaluation of red cell phagocytosis. PMID- 9407716 TI - Response of myelodysplastic syndrome bone marrow cells to multiple cytokine stimulation in liquid cultures: an in situ hybridization study. AB - BACKGROUND AND OBJECTIVE: Myelodysplastic syndrome progenitor cells can be grown and expanded in long term bone marrow liquid cultures in the presence of multiple cytokines. In this study we investigated the pattern of differentiation and response to growth factors in six cases of myelodysplastic syndrome (MDS) with well-defined cytogenetic abnormalities by means of conventional cytogenetics and fluorescence in situ hybridization (FISH). METHODS: Bone marrow cells were grown in stroma-free liquid cultures in the presence of SCF, IL-3, IL-6 and GM-CSF. RESULTS: IN three cases a CFU-GM expansion comparable to normal controls was observed, together with a decrease or increase of cells with abnormal karyotype. Two cases showed no response to growth factor stimulation, morphological signs of terminal myeloid differentiation and increase (one case) or decrease (one case) in the percentage of abnormal FISH signals along the cultures. In one additional case, while CFU-C expansion was present, clearcut leukemic transformation was observed in the culture, together with a sharp decrease in the percentage of abnormal FISH signals, indicating a leukemic transformation of MDS progenitor cells with a normal karyotype. INTERPRETATION AND CONCLUSIONS: Our data indicate that FISH analysis is generally a poor indicator of clonality in MDS; nevertheless, determining the kinetics of cytogenetically abnormal clones in liquid bone marrow cultures may provide insight as to the growth abnormalities of MDS progenitor cells and may be useful prior to in vivo growth factor administration. PMID- 9407717 TI - Clinical and biological characteristics of myelodysplastic syndromes with nulisomy Y by fish. AB - BACKGROUND AND OBJECTIVE: In the present study we analyzed the incidence of nulisomy Y by fluorescence in situ hybridization in a group of 24 males diagnosed with myelodysplastic syndromes (MDS). We explored the relationship between this chromosome abnormality and other clinical and biological disease characteristics. METHODS: Loss of chromosome Y was present in 7 out of the 24 males analyzed (29%); the number of cells carrying this chromosome aberration ranged between 19% and 90%. From the clinicobiological point of view, the group of patients with nulisomy Y showed a higher incidence of RA and RAS FAB subtypes (p = 0.04), a lower WBC count (p = 0.04), a lower proportion of blast cells both in PB (p = 0.009) and BM (p = 0.06) associated with a decreased myeloid/erythroid ratio (p = 0.01). RESULTS: No clear association was detected between loss of chromosome Y and other numerical chromosome abnormalities involving chromosomes 7 and 8. In contrast, 2 out of the 7 cases with loss of chromosome Y also displayed monosomy 1 by FISH. However, the use of appropriate dual stainings showed that these two abnormalities were present in different cell populations (that is, they never coexisted in the same cell population), which supports the notion of the existence of clonal heterogeneity in MDS patients. INTERPRETATION AND CONCLUSIONS: From the prognostic point of view, MDS patients with loss of chromosome Y displayed a higher survival rate, although these differences did not reach statistical significance. PMID- 9407718 TI - The patterns of IL2, IFN-gamma, IL4 and IL5 gene expression in Hodgkin's disease and reactive lymph nodes are similar. AB - BACKGROUND AND OBJECTIVE: The lymph nodes involved in classic Hodgkin's disease (HD), i.e. mixed cellularity (MC) and nodular sclerosis (NS) subtypes, usually contain few (1-2%) Reed-Sternberg (RS) cells scattered in a background of lymphocytes, eosinophils, plasma cells and neutrophils. CD4+ T-lymphocytes are increased in number, express activation markers and cluster around RS cells. The presence of eosinophilia in most HD patients and the presence of hyper-IgE in a subset of them may suggest that activated lymph node T cells release large amounts of IL5 and IL4, respectively. METHODS: The expression of four T-cell associated cytokine genes, i.e. interleukin (IL)2, IL4, IL5 and interferon (IFN) gamma, in frozen sections of 14 HD (7 MC, 7 NS) and 10 reactive lymph nodes was investigated by qualitative and quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). T-cell clones were also raised from purified CD4+ lymphocytes of one HD lymph node and one reactive lymph node and tested for IL2, IL4, IL5 and IFN-gamma secretion in culture supernatants by immunoassays. RESULTS: The transcripts of all the cytokine genes were detected in every lymph node irrespective of the HD or reactive nature. HD or reactive lymph node-derived CD4+ T-cell clones released the four cytokines according to a predominant T helper (Th)0-type pattern. In more than half of the lymph nodes of either HD or reactive nature, there was a predominance of IL4 over IFN-gamma mRNA production (Th2-type pattern). In the remaining HD or reactive lymphadenopathies, either a balanced IL4/IFN-gamma mRNA ratio (Th0-type pattern) or a predominance of IFN gamma over IL4 mRNA expression (Th1-type pattern) was observed. INTERPRETATION AND CONCLUSIONS: The overall pattern of cytokine gene expression in classic HD is similar to that detected in reactive lymph nodes. Further studies are needed to determine whether differences in the absolute concentrations of cytokines released in HD versus reactive lymph nodes and the long-standing course of HD versus the self-limiting nature of reactive adenopathies may explain certain peculiar features of HD, such as eosinophilia, for example. PMID- 9407719 TI - Mutational analysis of p53 in 16 cases of acute lymphoblastic leukemia and Burkitt's lymphoma. AB - BACKGROUND AND OBJECTIVE: Improvements in therapy for patients with B-cell acute lymphoblastic leukemia (ALL) and Burkitt's lymphoma (BL) depend on the identification of subsets of patients who require more intensive therapy. Abnormalities of the p53 gene are the most common molecular lesions in human cancer, and may be of prognostic significance in hematologic malignancies. In this study, we examined the p53 gene status in a group of patients with ALL/BL to determine whether some types of mutants were more frequent in this selected group of patients. METHODS: We selected a group of 16 patients with acute lymphoblastic leukemia (ALL) and Burkitt's lymphoma (BL) in order to investigate the presence of p53 mutations. DNA obtained from affected organs (bone marrow, lymph node and a renal mass) was used for the molecular studies. Single-strand conformation polymorphism (SSCP) analysis of exons 5 to 9 of the gene was used to detect p53 mutants. After detecting an abnormal migration pattern on the SSCP, mutations were determined by direct sequencing. RESULTS: Point mutations were found in eight patients; a misense mutation in seven cases and a non-sense mutation in one case. The normal allele was also identified in 7 mutated samples. The same mutation at codon 282 was identified in three different patients, in whom an identical conformer was detected after SSCP analysis. Mutation at codon 282 was present in an extramedular relapse (renal) appearing after a BMT. No such alteration was present in the bone marrow analyzed at the same time. INTERPRETATION AND CONCLUSIONS: Our findings suggest that p53 mutations are quite frequent in recognized clinical groups. The criteria chosen in this study allowed us to identify a high percentage of the samples with mutation. Different malignant phenotypes could be determined by functional heterogeneity of p53 mutants. PMID- 9407720 TI - Comparative flow cytometric evaluation of bcl-2 oncoprotein in CD5+ and CD5- B cell lymphoid chronic leukemias. AB - BACKGROUND AND OBJECTIVES: Levels of intracellular bcl-2 oncoprotein have been found to be increased in leukemic cells of CD5+ B-chronic lymphocytic leukemia (CLL) patients. However, it is not clear whether bcl-2 overexpression is a peculiar feature of CD5+ B-CLL. Based on this background we carried out a quantitative flow cytometric evaluation of intracellular bcl-2 levels on leukemic cells of CD5+ and CD5- B-CLL. METHODS: We assessed in flow cytometry levels of bcl-2 protein using a quantitative indirect immunofluorescence assay (QIFI kit) on samples from 46 previously untreated CD5+ B-CLL patients. Results were compared with those obtained on either normal peripheral blood B-lymphocytes or leukemic cells from 7 CD5- B-CLL patients intentionally selected for statistical comparison. RESULTS: A relatively homogeneous amount of bcl-2 protein which did not reflect either clinical-biological features at the time of diagnosis nor in vivo response to therapy was found. Results expressed as antibody binding capacity (ABC) accounted for a mean value of 12.2 +/- 1.5 x 10(3) molecules/cell (range, 6.4-13 x 10(3) molecules/cell). Levels of bcl-2 detected on CD5+ B-CLL leukemic cells were significantly lower than those of B peripheral blood lymphocytes from healthy donors (p = 0.0001). The same applied when comparing CD5+ and CD5- B-CLL patients (bcl-2 ABC, 8.07 +/- 0.26 x 10(3) molecules/cell vs. 12.2 +/- 1.5 x 10(9) molecules/cell; p = 0.0001). INTERPRETATION AND CONCLUSIONS: According to the role of bcl-2 in preventing apoptosis, our results indicate that differences in the pattern of expression of such an oncoprotein, might, at least in part, explain the more aggressive clinical course of CD5- B-CLL forms. PMID- 9407721 TI - The in vitro cytotoxic effect of mitoxantrone in combination with fludarabine or pentostatin in B-cell chronic lymphocytic leukemia. AB - BACKGROUND AND OBJECTIVE: Clinical studies indicate that combination chemotherapy with mitoxantrone (Mitox) and a purine analog can improve the response rate in indolent lymphoproliferative disorders. We explored the in vitro Mitox- fludarabine (FAMP)- and pentostatin (Pento)-induced cytotoxicity and their interactions in CLL. METHODS: The peripheral lymphocytes of 24 CLL patients were tested at different drug concentrations, with Mitox, FAMP or their combinations in 22 cases, and with Mitox, Pento or their combinations in 20 cases, 18 of which were the same from the FAMP group. The MTT assay was chosen for the drug-induced cell cytotoxicity and flow cytometry analysis of the DNA hypodiploid peak for the study of the apoptotic process. Drug interactions were calculated in the MTT assay according to both multiplicative and maximum models. RESULTS: According to the lethal dose (LD) 50 values, when the three drugs were tested alone, 11 out of 22 and 8 out of 20 samples were sensitive to Mitox in the FAMP and Pento groups, respectively; on the other hand, 2 out of 22 and 0 out of 20 samples appeared sensitive to FAMP or Pento alone, respectively. Analyzing the MTT assay data with the multiplicative and maximum model, the combinations of Mitox+FAMP and Mitox+Pento at different drug concentrations were synergistic in 28.2% and 39.3%, respectively. At leukemic cell survival < or = 50%, 11.7% and 11.1% of all combinations were synergistic in the Pento and FAMP group, respectively. The number of synergistic interactions at a therapeutically achievable plasma-drug concentration was an inverse function of the Mitox concentration. In the FAMP group, a direct correlation was found between the LD50 values of both FAMP and Mitox and the number of synergistic interactions, while the Pearson correlation coefficient was not significant in the Pento group. Finally, as measured by the DNA hypodiploid peak, Mitox (0.25 microgram/mL) plus Pento (0.16 microgram/mL) showed a significantly enhanced apoptosis in comparison to each single drug, while Mitox failed to demonstrate an additive effect with FAMP (1 microgram/ml). INTERPRETATION AND CONCLUSIONS: This experience demonstrates the extent of the in vitro synergism of Mitox with FAMP and Pento in inducing cell cytotoxicity; it also shows an adjunctive apoptotic effect for the Mitox-Pento association only. PMID- 9407722 TI - Inhibition of fibrin deposition on the subendothelium by a monoclonal antibody to polymorphonuclear leukocyte integrin CD11b. Studies in a flow system. AB - BACKGROUND AND OBJECTIVE: The development of prothrombotic and procoagulant states may be regulated by direct platelet-leukocyte contact mediated by membrane receptors. We have investigated the role of CD11b integrin in polymorphonuclear leukocytes (PMN) on fibrin formation and platelet reactivity with vascular subendothelium. METHODS: Studies were carried out following a well-established perfusion model, employing either citrated blood, where fibrin formation is blocked, or blood anticoagulated with low molecular weight heparin, which allows thrombin and fibrin formation. Isolated PMN or platelets were treated with specific monoclonal antibodies to CD11b or to CD62P, respectively, and incorporated in reconstituted blood. RESULTS: Treatment of PMN with anti-CD11b significantly decreased the percentage of surface covered with a thick layer (> 5 microns) of fibrin (34.8 +/- 3.3% vs 53.3 +/- 4.9% in control, p < 0.05); it also reduced the average height of fibrin layer and the number of adherent leukocytes (7.9 +/- 1.2 microns vs 10.6 +/- 1.4 microns in control, p < 0.05; and 87 +/- 8 PMN/mm2 vs 186 +/- 25 PMN/mm2 in control, p < 0.05) respectively. Treatment of PMN with CD11b did not significantly affect the attachment of platelets onto the subendothelium when using citrated blood, though a slight decrease in platelet adhesion was observed in the heparinized samples. Treatment of platelets with anti-CD62P did not significantly modify any of the parameters studied. INTERPRETATION AND CONCLUSIONS: Our results indicate that PMN have a role in promoting fibrin deposition under flow conditions, through the participation of CD11b integrin. Under our experimental conditions, this effect does not seem to be influenced by CD62P expressed on activated platelets. PMID- 9407723 TI - Clonal karyotype abnormalities in EBV-associated hemophagocytic syndrome. AB - BACKGROUND AND OBJECTIVE: An EBV-associated hemophagocytic syndrome (HS) in previously healthy children or young adults has been documented in Taiwan. The exact nature of this syndrome, i.e., either an infectious process or a neoplastic disease, remains to be clarified. METHODS: Three patients diagnosed as having HS were studied retrospectively. Chromosomes from bone marrow were examined by a conventional trypsin-Giemsa banding technique and karyotyped at the beginning of diagnosis or during treatment. In situ hybridization studies for EBV using EBER1 were performed. RESULTS: All three patients presented the classic manifestations of HS including fever, splenomegaly, jaundice, pancytopenia and coagulopathy. Bone marrow aspiration revealed atypical lymphocyte and histiocyte infiltration with hemophagocytosis. EBV genomes were found in bone marrow in all patients. In addition to normal mitotic cells, clonally karyotypically abnormal cells were demonstrated in all three patients whose diseases were rapidly progressive and eventually refractory to etoposide-based therapy. The consistent karyotypical abnormality of add(9)(p24) was noted in two of them. INTERPRETATION AND CONCLUSIONS: Although HS is usually considered a reactive process, the emergence of clonal cytogenetic abnormalities should be considered a malignant entity and treated with more intensive chemotherapy. A large series of cytogenetic and molecular studies is needed to clarify the exact nature of this fatal disease. PMID- 9407724 TI - The value of combination therapy in adult acute myeloid leukemia with central nervous system involvement. AB - BACKGROUND AND OBJECTIVE: In adult patients with acute myeloid leukemia (AML), central nervous system (CNS) involvement is a rare event and treatment has not yet been defined. Because there are no definitive data as to the most appropriate therapeutic approach to CNS leukemia in AML, we retrospectively analyzed a cohort of AML patients with meningeal leukemia in order to increase our knowledge on this particular matter. METHODS: Out of 410 patients with de novo AML observed at our Institute from 1986 to 1995, 9 (2.2%) showed CNS leukemia (CNSL) during the follow-up. CNSL was treated as follows: in a first group of 4 patients we combined systemic HD Ara-C 3 g/m2 (every 12 hours by 3-hour infusion, for 6 doses), cranial radiation therapy and intrathecal (IT) methotrexate (MTX); a second group of 4 patients was treated with HD Ara-C, IT MTX without cranial irradiation; HD Ara-C alone was administered in one patient. RESULTS: All patients of the first group and 2 patients of the second who achieved a complete remission (CR) had a median survival of 10 months (range 5-25+) after CNS involvement, while for the non-remitters it was 2 months (range 1-5). The only patient still living underwent allogeneic bone marrow transplantation. INTERPRETATION AND CONCLUSIONS: The combination treatment of HD Ara-C, IT MTX and cranial irradiation is well tolerated and seems to be an effective therapy for CNSL, presenting a high incidence of neurologic CR that correlates with a longer survival. As expected, the number of AML patients with CNSL was small, due to the fact that CNS in those patients is a rare complication. However, this study provides further information about the therapeutic possibilities in such restricted subsets of AML patients. PMID- 9407725 TI - Activation of the hemostatic process in patients with unruptured aortic aneurysm before and in the first week after surgical repair. AB - BACKGROUND AND OBJECTIVE: It has been previously suggested that activation of coagulation and fibrinolysis may sometime occur in patients with unruptured aortic aneurysm. However, the incidence of this complication and the effect of surgical repair are unknown. The objective of our study was to gain further information on this topic. METHODS: We investigated activation of the hemostatic process in 20 consecutive patients with unruptured abdominal aortic aneurysm. We then evaluated the effect of surgical repair of the vascular abnormalities. RESULTS: Both before and in the first week after surgery, the large majority of patients showed clear signs of activation of coagulation (increased plasma levels of prothrombin fragment 1 + 2 and fibrin peptide A), and many had low levels of the natural anticoagulant antithrombin III. Platelets were activated in all cases (high levels of plasma beta-thromboglobulin), and signs of platelet consumption (thrombocytopenia and/or increased mean platelet volume) were present in most of them. INTERPRETATION AND CONCLUSIONS: Activation of the hemostatic process occurs in nearly all patients with abdominal aortic aneurysm and could play a role in the hemorrhagic and thrombotic events that can complicate the clinical development of these subjects. PMID- 9407726 TI - Pilot study on the safety and efficacy of desmopressin for the treatment or prevention of bleeding in patients with hematologic malignancies. AB - BACKGROUND AND OBJECTIVES: Desmopressin is the treatment of choice for patients with von Willebrand's disease and mild hemophilia A. This compound is also useful in other congenital and acquired disorders of hemostasis, reducing the need for blood derivatives with the inherent risks of infections and alloimmunization. The following article presents a pilot study on the safety and efficacy of desmopressin for the treatment or prevention of bleeding in 15 patients with thrombocytopenia associated with hematologic malignancies. METHODS: Cases were consecutively recruited from February to June 1995. Fifteen patients were treated with desmopressin for prevention or treatment of bleeding. Desmopressin was diluted in 100 mL of isotonic saline and infused for 30 minutes. Bleeding time (BT) was carried out using the Simplate II device, making two standardized incisions on the forearm: the mean between the two incisions was recorded. RESULTS: Significant reduction of BT was observed in three out of four patients with myelodysplastic syndrome who were successfully treated for active bleeding or dental extraction. In the remaining patients, the effect of desmopressin on BT was not tested. Nevertheless, in all of them bleeding mainly due to epistaxis or persistent gum oozing was stopped by a single infusion of desmopressin. In three patients, desmopressin infusion had been successfully administered on a different occasion. No side effects were observed. INTERPRETATION AND CONCLUSIONS: Desmopressin could be a safe and immediately effective option for the treatment or prevention of bleeding in selected patients with hematologic malignancies. PMID- 9407727 TI - Chimerism analysis in long-term survivor patients after bone marrow transplantation for severe aplastic anemia. AB - BACKGROUND AND OBJECTIVE: Allogeneic bone marrow transplantation (BMT) is the most common treatment for young patients with severe aplastic anemia (SAA). Late graft failure represents one of the possible unfavorable outcomes in this setting. Mixed chimerism might represent a risk factor for late graft failure. We examined this relationship by studying chimerism in long-term survivor SAA patients after allogeneic BMT. METHODS: We analyzed long-term hematopoietic chimerism in 15 patients who received BMTs for SAA: 9 with an irradiation-based conditioning regimen and 6 with ATG. We used a PCR method targeting VNTR loci. Sensitivity of the technique ranged between 0.5 and 1.5%. RESULTS: All patients conditioned with radiation-based schemes showed complete donor chimerism. Conversely, out of six patients who received cyclophosphamide and ATG as a conditioning regimen, only one of them had late graft failure (day +168). In this patient, durable mixed chimera status was first detected two months after BMT. INTERPRETATION AND CONCLUSIONS: Our results suggest that in long-term survivors of SAA after BMT there is almost always complete donor chimerism in both irradiated and ATG-conditioned recipients. Mixed chimerism might predict graft failure in these patients. PMID- 9407728 TI - Feasibility of molecular diagnosis of alpha-thalassemia in the evaluation of microcytosis. AB - Microcytosis is a common hematological finding, usually related to iron deficiency or beta-thalassemia. When both of these conditions are excluded, alpha thalassemia must be considered in the differential diagnosis. No simple biochemical test is able to diagnose the alpha-thalassemia trait. Using PCR amplification of the breakpoint in deletional forms, and amplification of the alpha 2 gene and restriction enzyme digestion in non-deletional forms, we identified the alpha-thalassemia carrier status in 42 out of 51 (82%) patients with microcytosis or slight microcytic anemia, unrelated to iron deficiency or beta-thalassemia. Our results underline the usefulness of molecular tests in clinical practice. PMID- 9407729 TI - Bone marrow involvement in lymphoblastic lymphoma and small non-cleaved cell lymphoma: the role of trephine biopsy. AB - Trephine biopsy (TB) combined with bone marrow aspiration (BMA) is the most common method for evaluating bone marrow (BM) involvement in non-Hodgkin's lymphomas. Nevertheless, the role of TB in high-grade lymphomas remains controversial. We reviewed the results of 42 consecutive BMAs and TBs performed simultaneously in 29 patients with lymphoblastic lymphoma (LL) and small, non cleaved cell lymphoma (SNCL). In LL, 8M involvement was documented in 35.4% of the cases by BMA and 22.5% of the cases by TB. In SNCL it was documented in 45.4% of the cases by BMA and 36.3% by TB. There were no statistically significant differences (p > 0.05) in the rates of BM involvement found by TB or BMA in the two types of lymphoma, although BMA appeared to be more sensitive than TB. These observations suggest that routine TB may not be necessary in assessing BM involvement in patients with LL and SNCL. PMID- 9407730 TI - Simultaneous occurrence of B-cell chronic lymphocytic leukemia and chronic myeloid leukemia with further evolution to lymphoid blast crisis. AB - The coexistence of chronic myeloid leukemia (CML) and B-cell chronic lymphocytic leukemia (CLL) in the same patient is rare. A 71-year-old woman developed a B lineage lymphoid blast crisis at 18 months after diagnosis of Ph-positive CML. At this time, a lymphoid cell population with morphologic and immunophenotypic features of CLL was demonstrated. The retrospective review of the tests performed at diagnosis and thereafter disclosed the presence of lymphoid nodules in the initial bone marrow biopsy in the absence of lymphocytosis. Subsequently, there was an appearance of moderate lymphocytosis in the following months. Therefore, diagnosis of CML and coexistent CLL was established. Although a transient remission of blast crisis was achieved, blast cells reappeared two months later and the patient died shortly afterwards. Molecular studies of the immunoglobulin heavy chain gene (IH) rearrangement pattern point to the origin of the diseases in two different cell clones. In addition, previously published cases of simultaneous CLL and CML are reviewed. PMID- 9407731 TI - Use of a rapid method for genotyping human platelet antigen systems in neonatal alloimmune thrombocytopenia. AB - We applied a polymerase chain reaction-sequence specific primer (PCR-SSP) method developed by other researchers to study 4 families of newborns with neonatal alloimmune thrombocytopenia (NAITP) in which serology had provided inconclusive human platelet antigen (HPA) typing data. This method allowed for the identification of the newborn HPAs which were incompatible with their respective mothers. They were HPA-2b, -1b, -3a, and -5b. This PCR-SSP is a useful tool for improving the ability to identify the incompatible HPA in NAITP. PMID- 9407732 TI - Immune thrombocytopenic purpura secondary to endometriosis. AB - In this paper, we describe a patient with immune thrombocytopenic purpura (ITP) secondary to endometriosis. To our knowledge, this is the first reported case presenting such an association. Surgical eradication of the endometriosis was the only effective treatment for the thrombocytopenia. The pathogenic connection between both disorders seems to be an altered immune function. PMID- 9407733 TI - Complete cytogenetic but not molecular remission in a patient with myeloid blast crisis of chronic myeloid leukemia treated with carboplatin and ara-C. AB - We report on a patient diagnosed with myeloid BC-CML in which a complete cytogenetic remission confirmed by FISH assay was obtained after therapy with carboplatin-ARA-C. However, RT-PCR analysis showed persistence of the p210 bcrabl translocation. Accordingly, the level of residual malignant cells should be between 10(-2) and 10(-6). Autologous stem cell transplantation was performed, but relapse occurred 11 months after blast crisis. This case supports the effectiveness of a carboplatin-ARA-C protocol in BC-CML in order to induce cytogenetic remissions. PMID- 9407734 TI - Effects of recombinant human granulocyte colony-stimulating factor administration on neutrophil phenotype and functions. AB - BACKGROUND AND OBJECTIVE: Recombinant human granulocyte colony-stimulating factor (rhG-CSF) is currently used for treatment of various types of neutropenia, treatment of aplastic anemia, mobilization of peripheral blood progenitor cells. However, rhG-CSF is not only a growth factor for the myeloid lineage, but it also acts as a modulator of neutrophil behavior. The aim of the present review article is to examine the following aspects of rhG-CSF therapy: 1) does rhG-CSF influence neutrophil functions, and in particular their microbicidal properties? 2) does rhG-CSF modify neutrophil phenotype? 3) If so, what are the mechanisms potentially involved? EVIDENCE AND INFORMATION SOURCES: The author of the present article has been working in the field of rhG-CSF effects on neutrophil function, contributing original papers in peer-reviewed journals. In addition, the present review critically examines articles and abstracts published in journals covered by the Science Citation Index and Medline. STATE OF ART AND PERSPECTIVES: Treatment with rhG-CSF causes enhancement of functions such as phagocytosis, superoxide anion generation, chemiluminescence, bacterial killing, and ADCC. Neutrophil phenotype changes after rhG-CSF administration: immediate effects cause direct activation of circulating neutrophils, but delayed effects are characterized by increased surface expression of important effector molecules directly involved in neutrophil functions, such as CD14, CD32, CD64. These effects may have useful clinical consequence in patients who show an increased risk of infections, such as cancer patients, subjects with hematologic diseases (myelodysplasia, aplastic anemia), congenital diseases characterized by neutropenia, and patients with AIDS. Other changes which characterize neutrophils after rhG-CSF administration are represented by significant impairment of CD16 expression, chemotaxis, and reduced in vivo migration of neutrophils to inflammatory sites. These effects may be explained by bone marrow modification due to rhG-CSF therapy. In fact, treatment with rhG-CSF causes a significant acceleration of transit time of cells belonging to the myeloid lineage, along with amplification of the mitotic pool and a relative decrease of elements of the post-mitotic pool. It is possible that, because of the accelerated bone marrow transit time of myeloid cells, rhG-CSF causes a relative immaturity of circulating neutrophils. It is known that both CD16 expression and chemotaxis properties are acquired by neutrophils in the late stages of maturation, but the time necessary to acquire full functional maturity seems to be shortened by rhG CSF administration, and this kinetic aspect may play a non-negligible role in the modification of neutrophil behavior. PMID- 9407735 TI - Expression of the stem cell factor receptor C-KIT (CD117) in acute leukemias. AB - BACKGROUND AND OBJECTIVE: The receptor for stem cell factor (CD117) is the gene product of the c-kit proto-ontogene. Together with its ligand, the stem cell factor (SCF), it plays an important role in hematopoiesis. In this study, we review the cellular distribution of CD117 in normal hematopoiesis and in hematopoietic malignancies focusing on the differential expression in subtypes of acute leukemias. EVIDENCE AND INFORMATION SOURCES: This review is based on a literature search in the Medline database, personal publications and results obtained as a reference laboratory of the German AML-BFM, AMLCG and ALL multi center therapy studies. STATE OF THE ART AND PERSPECTIVES: Membrane expression of CD117 can be found on leukemic blasts from approximately 60% of adult and childhood AML patients, often associated with an immature immunophenotype (CD34). Moreover, AML with t(8;21) are frequently CD117 positive. Despite earlier reports, most recent studies have not been able to demonstrate any significant prognostic impact of CD117 expression in either childhood or adult AML. A small proportion of T-lineage ALL (9%), mainly consisting of immature pro-T/pre-T-ALL, is CD117 positive. CD117 expression is rare in B-cell-precursor-ALL and occurs in less than 3% of cases. CD117 in combination with other antigens might facilitate the immunologic characterization of acute leukemias, especially those of myeloid and early T-cell origin. PMID- 9407736 TI - Cytokine receptors, growth factors and cell cycle in human bone marrow and peripheral blood hematopoietic progenitors. AB - BACKGROUND AND OBJECTIVE: An increasing number of growth factors have been shown to be responsible for the proliferation, survival and enhanced function of many cell types within the hemopoietic system. The action of these hemopoietic growth factors in stimulating cell growth and survival applies both to cells within the progenitor compartment and mature cells. Whether a specific cytokine influences in vivo hematopoietic progenitor cell proliferation or survival depends on cytokine-mediated modulation or target cell cytokine receptors, cell proliferation, and cell death regulator genes and other pathways. To address these issues, particularly in view of the current and future clinical use of hemopoietic growth factors, the Italian Society of Experimental Hematology organized a Meeting in Florence on July 4th, 1996. INFORMATION SOURCES: The material examined in the present review includes full papers and abstracts published in journals covered by the Science Citation Index and Medline. All the participants to the Meeting in Florence have been actively working in the field of biology and clinical application of hemopoietic growth factors. Summaries of their oral presentations at the Florence Meeting are reported in the Appendix to this article. STATE OF ART AND PERSPECTIVES: Myelopoietic growth factors particularly granulocyte (G-) colony-stimulating factor (CSF) and granulocyte macrophage (GM)-CSF, have been available for clinical use for only a few years but they have already markedly changed the management of chemotherapy-induced neutropenia, the use of dose-intensive chemotherapy regimens and the practice and safety of autologous stem cell transplantation. While these growth factors have been rapidly introduced as routine agents in the management of cancer patients, they have continued to generate a considerable amount of fundamental research into the biology of hematopoiesis as well as the growth regulation of normal and cancer cells. For instance, one goal of cancer treatment is to protect hematopoietic stem and progenitor cells from the damaging effects of chemotherapy, while maintaining their anticancer action. Any means of preferentially and reversibly suppressing the proliferation of normal hematopoietic stem and progenitor cells while leaving the proliferation of tumor cells and their susceptibility to chemotherapy unmodified, could potentially optimize treatment efficacy. In this field, the possibility of using colony stimulating factors as myeloprotective agents in dose-intensive chemotherapy to enhance anticancer activity could be an attractive goal of current anti-cancer treatment modalities. PMID- 9407737 TI - Use of plasmas from donors under oral anticoagulant treatment for the expression of INR values. PMID- 9407738 TI - Dendritic cell differentiation in a peripheral blood. Mononucleated cell culture treated with interleukin-2. PMID- 9407739 TI - The syndrome of abnormal chromatin clumping in leukocytes. PMID- 9407740 TI - Central nervous system (CNS) infiltration in a case of promyelocytic leukemia. PMID- 9407741 TI - Warfarin resistance induced by teicoplanin. PMID- 9407742 TI - Early autologous stem cell transplantation in Hodgkin disease in partial remission or in relapse. PMID- 9407743 TI - Probable ticlopidine-induced severe aplastic anemia and cholestatic hepatitis. PMID- 9407744 TI - Desferrioxamine in the treatment of myelodysplastic syndromes. PMID- 9407745 TI - Effects of football training on ventilatory and gas exchange kinetics to sinusoidal work load. AB - The purpose of this study was to evaluate the effects of football training on maximal oxygen uptake (VO2max), ventilatory threshold (VT) and kinetics of ventilation and gas exchange variables to sinusoidal work load. The sinusoidal work load during cycling exercise was varied from 30 watts to 60% of VO2max (60% VO2max) with a period of 2 min. O2 uptake (VO2), CO2 output (VCO2), minute ventilation (VE) and heart rate (HR) were measured on a breath-by-breath basis using a computer system. Training periods were continued for 9 months. Six males who had no football experience were performed in which VO2max, VT, and the kinetics of each variable were measured for 3-month intervals (PRE, TR.3, TR.6, and TR.9). They usually underwent football training, such as sprint and strength training for 2-3 hours day-1, 6 days week-1. Mean VO2max was significantly increased at TR.3 and TR.6. VT was also significantly increased during the training period. The amplitude of VO2, VCO2, VE, and HR responses during sinusoidal exercise unchanged during the training period. Phase shifts to work load in VO2 and HR responses did not significantly change during the 9 months, but the phase shift in VCO2 and VE responses significantly continued increasing as the training intervals progressed. These results suggest that football training does not significantly affect the development of the kinetics of VO2 and HR during submaximal exercise, but that it dramatically increases VO2max and VT values. PMID- 9407746 TI - Oxygen consumption and energy expenditure of level versus downhill running. AB - OBJECTIVE: The purpose of this study was to assess and compare submaximal oxygen consumption (VO2) and energy expenditure (kJ) while running at 0, -1.8, -3.6, and -5.4% grades for three individually selected running speeds (9.4 + 0.79, 10.3 + 0.74, 11.3 + 0.73 km.h-1). EXPERIMENTAL DESIGN: Subjects completed the four grade conditions in random order via a modified Latin squares design at three self selected submaximal running speeds for each condition. PARTICIPANTS: Thirteen (5 females and 8 males) recreational (< 35 km.wk-1) runners (age: 27.7 +/- 4.3 yrs) volunteered for the study. MEASURES: Two-way repeated measures ANOVA (Grade x Speed) was used to analyze steady-state VO2 and kJ expenditure. Stepwise linear multiple regression was used to develop an equation for predicting VO2 for running at recreational speeds on moderately negative grades. RESULTS: VO2 and kJ mean values were significantly different between all speed and % grade comparisons. Compared to level grade, the average reductions in VO2 and kJ expenditure ranged from approximately 9% at a grade of -1.8% to 22% at a grade of -5.4%. The relationship between VO2 and % grade for each running speed was linear. CONCLUSIONS: For a given speed, running at a modest negative grade can significantly decrease VO2 and kJ expenditure compared to level running. The following regression equation can be used to estimate VO2 (ml.kg-1.min-1) for running at recreational speeds on slight downhills: VO2 = 6.8192 + 0.1313 (speed in m.min-1) + 1.2367 (% grade). PMID- 9407747 TI - Effects of deep water and treadmill running on oxygen uptake and energy expenditure in seasonally trained cross country runners. AB - OBJECTIVE: The purpose of this study was to physiologically compare submaximal intensity deep water running (DWR) and treadmill running (TMR) exercise in trained athletes. EXPERIMENTAL DESIGN: Pre-test, post-test, 2 x 2 factoral design. SETTING: Treadmill exercise tests occurred in the Human Performance Laboratory. DWR trials took place in the deep end of the University pool. PARTICIPANTS: Seasonally trained college-aged male cross country runners (N = 8). Subjects completed a treadmill maximal oxygen consumption (VO2max) test, followed by a submaximal treadmill and deep water run at heart rates equivalent to 60% and 80% treadmill VO2max. MEASURES: Oxygen consumption (VO2), ventilation (VEstpd), rates of perceived exertion (RPE), energy expenditure (kcal.min-1), respiratory exchange ratio (RER), fat and carbohydrate oxidation (g.min-1) were measured during two 5 minute steady state stages for both trials. RESULTS: The trial by intensity interaction for VEstpd was significant, demonstrating greater ventilation during DWR as compared to TMR at 80% VO2max. The main effect of trial demonstrated that significantly higher RER and carbohydrate oxidation, and lower fat oxidation occurred during DWR as compared to TMR. VO2, RPE, and energy expenditure did not differ significantly between trials. CONCLUSIONS: DWR is a comparable form of submaximal intensity exercise as TMR in well-trained athletes. DWR does, however, maintain unique properties that differs it from TMR. Therefore, the concept of training specificity should be further considered when prescribing DWR and using it as an enhancement tool or substitute for dry land running. PMID- 9407748 TI - Cardiorespiratory response to exercise in patients with exercise-induced bronchial obstruction. AB - OBJECTIVE: To document the characteristics of the ventilatory response to exercise in patients with exercise-induced bronchial obstruction (EIB). EXPERIMENTAL DESIGN: Comparative study during the period between December 1993 and March 1994. SETTING: Ambulatory care in Ohgaki Municipal Hospital. SUBJECTS AND METHOD: We evaluated 11 children with EIB. Each subject under went symptom limited cardiopulmonary treadmill exercise testing (Bruce protocol). RESULTS: Patients with EIB showed a significantly lower peak oxygen consumption (peak VO2) than the control subjects. Three patients with EIB developed relative hypoventilation during incremental exercise: an increase in end-tidal carbon dioxide partial pressure (PETCO2) and a decrease in VE/VCO2 were observed at the end-stage of exercise testing. CONCLUSIONS: These findings demonstrate that some patients with EIB develop bronchoconstriction during exercise. PMID- 9407749 TI - Swimming performances and stroking parameters in non skilled grammar school pupils: relation with age, gender and some anthropometric characteristics. AB - OBJECTIVE: It was hypothesized that swimming velocity (V) and stroking parameters such as stroke length (SL), stroke rate (SR) and stroke index (SI) are influenced by age, gender, and some anthropometric characteristics. EXPERIMENTAL DESIGN: Cross-sectional study. SETTING: Grammar school pupils from French schools. PARTICIPANTS: One thousand and ninety-seven males and 961 females non skilled swimmers aged from 11 to 17. INTERVENTIONS: Usual swimming sessions (6 +/- 2 h.year-1) during a physical education program at school. MEASURES: V, SL, SF and SI (SI = V.SL) were measured or calculated from a 50-m crawl sprint and height, arm span and body mass were measured for all subjects. RESULTS: The results showed that V, SL and SI increased regularly (p < 0.01) in relation to age in both genders. SL was never significantly different between males and females. SF remained unchanged according to age and was significantly higher in males than in females. V, SL and SI were influenced by age and arm span but not SF. CONCLUSIONS: As males and females were submitted to the same swimming teaching program at school, a higher increase in muscle power and anaerobic capacity in males could explain the gender differences. These results observed in non skilled swimmers showed that the differences in stroking parameters between genders were the reverse of those of top level swimmers and that they can be used by swimming teachers in order to build some assessment tools and to better understand the improvements in swimming in relation to growth and gender. PMID- 9407750 TI - May peripheral and central fatigue be correlated? Can we monitor them by means of clinical laboratory tools? AB - BACKGROUND: A laboratory-based model which links regional and central fatigue during physical exercise has not yet developed. Today we can assay the oxypurines, a specific and sensible marker of muscle cell-energy exhaustion during strenuous physical exercise, thus allowing us to insight in peripheral fatigue mechanisms. Prolonged physical exercise modifies plasma free amino acids and fatty acids levels, increases plasma free tryptophan (fTrp) and, conversely, probably serotonin, an amine involved in the genesis of central fatigue. We tried to verify if there is a correlation between central and peripheral fatigue. EXPERIMENTAL DESIGN: We studied 29 male marathon runners before marathon, at the arrival, one and three days after the run. MEASURES: Plasma samples were assayed for amino acids, fTrp serotonin, xanthine, hypoxanthine inosine, cortisol. Urine samples were assayed for serotonin and hydroxyin-doleacetic acid (5HIAA). RESULTS: After the competition we observed a decrease in plasma fTrp but an increased ratio fTrp/sum of neutral amino acids with a normalization after 24 hours. No significant changes were observed in plasma and urinary serotonin and 5HIAA. Hypoxanthine and inosine increased at the end of the trial and returned to basal levels the day after. Cortisol increased at the end of the run but was reduced after 24 and 72 hours. CONCLUSIONS: In our athletes we observed only indirect signs of fTrp involvement in the genesis of central fatigue. Oxypurines seem to be a good marker of regional muscular fatigue. Plasma cortisol expresses the stress reaction to the competition and its exhaustion after a prolonged physical exercise. PMID- 9407751 TI - Effect of exercise intensity on mood in step aerobics. AB - BACKGROUND: The purpose of this study was to examine how aerobic exercise and exercise intensity affect transient mood states. A two-level (high intensity exercise/low intensity exercise) pretest-post-test design was employed. METHODS: Subjects included 42 healthy male and female volunteers (M age = 39.12; SD = 11.53, range 17-64 years) enrolled in four aerobics classes. The classes were randomly assigned to either low intensity (< or = 60% maximum heart rate) or high intensity (> or = 75% maximum heart rate) conditions. Subjects participated in a 50-minute bench-stepping routine at their assigned intensity level. Pre-exercise and post-exercise assessment of transient mood was assessed with the Profile of Mood States (POMS) inventory. RESULTS AND CONCLUSIONS: Results from a 2 x 2 (pre exercise/post-exercise x high intensity/low intensity) ANOVA suggested that tension, depression, fatigue and anger decreased while vigor increased in both conditions. Additionally, subjects in the high intensity group reported less fatigue and anger than those participants in the low intensity group. PMID- 9407752 TI - Affective responses to acute exercise: a test of opponent-process theory. AB - Based on Solomon's Opponent-Process theory (1980), it was predicted that individuals involved in a regimen of regular aerobic exercise (active; n = 18) would respond to an acute bout of exercise with reduced negative and/or increased positive affect compared to nonactive counterparts (nonactive; n = 12). State Anxiety (SA), positive affect (PA), negative affect (NA), and self-reported fatigue were assessed immediately prior to, every 6 min during, and every 6 min following a 24 min bout of bicycle exercise performed at an RPE of 13 (+/- 1). As expected, no significant group differences occurred for RPE (M = 13.5 for nonactive, 13.2 for active). The active group did, however, exercise at a greater absolute workload than the nonactive group (261.0 +/- 22.4 W vs 200.0 +/- 19.98 W, respectively). Analyses indicated similar changes in SA and fatigue for both groups, with significant reductions in SA occurring at 6 min post-exercise and remaining below pre-exercise levels throughout the post-exercise period, while fatigue was reduced at 12, 18, and 24 min post-exercise. A significant Group x Time interaction occurred for affective valence (PA - NA; p < .01). Post hoc analyses indicated that for the active group, affect increased modestly (i.e., more PA, less NA) during exercise; this increase was sustained post-exercise. The nonactive group evidenced a sharp drop in affect (i.e., less PA, more NA) during exercise followed by a small post-exercise rise which did not return to pre exercise levels. The results of the present study partially support the Opponent Process model as an explanation for exercise-related affect. Although there was no differential anxiety response as a function of activity level as the model would predict, there was a differential response for affective valence in accordance with predictions. PMID- 9407753 TI - Knowledge and reported use of sport science by elite New Zealand Olympic class sailors. AB - OBJECTIVE: This study enquired about the knowledge and reported use of sport science in elite Olympic class sailors. EXPERIMENTAL DESIGN: The sailors responded to a simple questionnaire. SETTING: The questionnaire was administered as part of an introductory seminar on sport science during a training camp. PARTICIPANTS: The participants were 28 (22 male, 6 female) elite New Zealand Olympic class sailors. INTERVENTIONS: None. MEASURES: The questionnaire asked whether or not they used a training race diary, enquired about their current and past injuries and their knowledge and use of sport science in the areas of nutrition, psychology and physical training. RESULTS: Only ten (36%) of the sailors kept a training/race diary. Whilst only four (14%) had a current injury, sixteen (57%) reported an injury in the previous three years. The injuries were in the lower back (45%), knee (22%), shoulder (18%), and arm (15%). Although nineteen (68%) of the sailors had experienced dehydration during racing, the average volume of fluid reported to be taken on a four hour sail was only 0.9 litre, of which only an average of 0.7 litres (77%) was reported to be drunk. All the sailors reported being sometimes (46%) to very often (3%) anxious before races and sometimes (43%) to always (7%) being frustrated with their own mistakes. Only one sailor reported never having negative thoughts whilst fifteen (53%) reported having them sometimes, and seven (25%) often or very often. Twenty four (86%) of the sailors reported that they sometimes had a loss of concentration near the end of the race. Whilst eighteen (64%) reported practising relaxation and seventeen (61%) reported practising visualisation as a mental skill, only five (18%) practised progressive mental relaxation, two (7%) practised meditation and none practised yoga. Seventeen (61%) undertook strength/circuit training, ten (36%) flexibility and twenty-one (75%) off water aerobic training. Twenty-four (86%) reported undertaking on-water aerobic training. CONCLUSIONS: The results indicate that there is considerable scope for improvement in the knowledge and use of sports science amongst elite New Zealand Olympic class sailors. PMID- 9407754 TI - Epidemiologic approach of doping in sport. A review. AB - OBJECTIVE: To determine, the prevalence of doping in sport as it was reported by the athletes during surveys, and to try to isolate risk factors to resort to doping. EXPERIMENTAL DESIGN: Medline, Pascal and Embase search for the period from 1980 to 1996. STUDY SELECTION: Of the 44 studies produced, 15 were not included in the detailed data summary because they reported statistical data from antidoping controls, they were not enough specific, or they concerned horse races. DATA EXTRACTION: Details of study design, drugs studied, prevalence. When available, were also noted: the sport practiced and the motives for doping. RESULTS: Among children, doping prevalence in around 3-5%. Among adults, in self reported use studies, doping prevalence may be estimated at 5-15%, where projected use studies report a mean prevalence near 15-25%. Studies provide few data about the sports that produce drug users. CONCLUSIONS: The extent of sport doping and its potential risks for health must make it to be considered as a problem of public health. This means that physicians must, at last, consider it as any other problem and change their behaviour against doping, in order not to reduce the subject to the sole list of prohibited substances. As for sports federations, they must, as last, recognize that prevalence of doping is high. Lastly, new studies are essential to determine motives for doping and to institute the predictable factors for this practice, what will perhaps make efficient the prevention campaigns. PMID- 9407755 TI - Stress fracture of the ulna in a female table tennis tournament player. AB - A stress fracture of the diaphyseal ulna occurred in a 19-year-old female competitive table tennis player with a history of secondary amenorrhea. Stress fractures are one of more common problems occurring in vigorous sports. Stress fractures of the ulna can result from any repeated forearm flexor muscle activity. In our case stress was caused by increased athletic activity by additional training twice a day six times a week starting eighth weeks before first signs of pain and by rhythmic repeated submaximal work load in a new job additional to training. Physical training is often associated with menstrual irregularity. Hormonal factors were known for six months. Spontaneous onset of pain in women and previous history of amenorrhea should include stress fractures as differential diagnosis. PMID- 9407757 TI - A brave STEP forward--the Singapore Tuberculosis Elimination Programme. PMID- 9407758 TI - Abdominal tuberculosis. PMID- 9407756 TI - Diclofenac dispersible provides superior analgesia with faster onset of action compared to naproxen granular in patients with acute, painful, minor sports injuries. AB - BACKGROUND: To compare the efficacy and tolerability of single doses of diclofenac dispersible and naproxen granular in patients with acute, painful, minor sports injuries. METHODS: Forty-eight adult outpatients with moderate-to severe pain on movement, following a traumatic event < or = 36 hours previously, participated in this double-blind, between-patient comparative study. Patients were randomised in equal comparative study. Patients were randomised in equal number to receive diclofenac dispersible 50 mg or naproxen granular 500 mg. Pain on movement, pain on pressure, spontaneous pain and pain relief were assessed at 15, 30, 45, 60 minutes and 4 hours after dosing. RESULTS: Both treatments were effective at reducing pain from the 15 minute time point. At 15 minutes there was no significant difference between the treatments for pain on movement (p = 0.4) but diclofenac was significantly superior to naproxen with respect to pain on pressure (p = 0.004), spontaneous pain (p = 0.0022) and pain relief (p = 0.034). In addition, diclofenac was significantly superior to naproxen with respect to AUC0-4 hours for percentage reduction in intensity of pain on movement (p = 0.04) and spontaneous pain (p = 0.0047), and for pain relief scores (p = 0.015). Both treatments were well tolerated. CONCLUSIONS: The study results suggest that diclofenac dispersible 50 mg provides faster and overall better analgesia compared to naproxen granular 500 mg in the acute relief of pain following minor sports injuries. PMID- 9407759 TI - Abdominal tuberculosis: a presumptive diagnosis. AB - BACKGROUND: Abdominal tuberculosis (TB) is common. But the diagnosis of abdominal TB is fraught with difficulties as it is often not possible to get a microbiological confirmation of the infection. We therefore undertook this study to highlight those pertinent clinical and laboratory features which would enable one to make a provisional diagnosis of abdominal TB early, to pave way for a trial of anti-tuberculosis chemotherapy. METHOD: This is a retrospective study of 12 patients treated for abdominal TB in our department over a period of 2 years. FINDINGS: Seven of the patients suffered from chronic diarrhoea for periods ranging from 4 weeks to 12 months. Four patients had progressive abdominal distension (ascites). The last patient came in with multiple abdominal swellings. Seven patients had clinical and biochemical features of malabsorption. Another 9 patients also had persistent pyrexia. The ascitic fluid was exudative in the 4 patients mentioned earlier. A definitive diagnosis could not be established in any of these patients. The diagnosis of abdominal TB was thus one of exclusion in these patients who showed prompt response to anti-tuberculosis therapy. CONCLUSION: Our study justifies a trial of anti-TB chemotherapy in TB endemic areas in the following clinical situations: (a) patients with chronic diarrhoea of unknown aetiology and (b) patients with exudative ascitic fluid, after all other possible causes, have been excluded. A prompt response to anti-TB therapy should be accepted as sufficient ground for the diagnosis of abdominal TB even when histopathological or microbiological confirmation of the disease is not possible. Our study reflects the experience of other workers from Third World countries. PMID- 9407760 TI - Diagnosis of ectopic pregnancy--why we need a protocol. AB - OBJECTIVE: To audit the management after instituting a screening programme for ectopic pregnancy in an institution with a protocol utilising ultrasound examination and serial human chorionic gonadotropin (hCG) and to examine the risk of missed diagnosis with deviation from the protocol. MATERIAL AND METHOD: A retrospective analysis of the management of 145 symptomatic patients in early pregnancies without intrauterine gestational sacs from ultrasound examinations, during the period April to June 1994 in Kandang Kerbau Hospital. Patients underwent serial hCG tests over 48 hours with or without repeat ultrasound scans before definitive treatment unless clinical indications for emergency surgery was necessary. RESULTS: There were 35 ectopic pregnancies (24%), 16 were viable intrauterine pregnancies (11%), 87 were non-viable pregnancies (60%) and 7 were of unknown outcome. There were much practice deviations from the protocol. Forty four percent (64 cases) of the management decisions were made based on the initial clinical and ultrasound findings, and another 14% (21 cases) after a repeat assessment within the next day by either a repeat scan or serial serum hCG over one day. Among them, two of the 29 operated for suspected ectopic pregnancy were not ectopic (7%) and two of the 56 thought not to be ectopic, turned out to be ectopic (4%) (p < 10(-8)). Six percent (8 cases) defaulted after the initial assessments and one of them was found to be ectopic subsequently. Thirty percent (43 cases) adhered to the protocol. They had serial serum hCG done over two days. Seven of them requiring further repeats of serial serum hCG before management decisions were made. Four patients who were operated on were confirmed ectopic and 39 patients not operated on were not ectopic. Three percent (5 cases) were managed by serial hCG over 3 to 5 days and another 3% (4 cases) by repeating scan over one to two weeks without serial hCG. None of these was ectopic. The percentage change of hCG levels over two days gave indications of the likely diagnosis. CONCLUSION: Adhering to a protocol utilising the principle of ultrasound scan, serial hCGs and selective repeat ultrasound scans are highly recommended for the diagnosis of ectopic pregnancy. Any deviation from protocol is dangerous, with a 4% risk of missing an ectopic and a 7% risk of unnecessary operation for suspected ectopic pregnancy. PMID- 9407761 TI - Acute pain service, Kandang Kerbau Hospital, 1995--a first year's experience. AB - AIM: To determine the efficacy and safety of the different modes of post operative analgesia in the first 24 hours for both caesarean section and major gynaecological procedures. METHODS: Being a purely descriptive study, the patients were not randomised to the post-operative analgesic mode. The choice was made both by the patient and the anaesthetist. The modes employed were continuous intravenous morphine, continuous intravenous pethidine, epidural morphine/bupivacaine, epidural bupivacaine/fentanyl and spinal morphine. During the first 24 hours, the patients' maximum pain scores, ability to sleep well, presence of nausea and/or vomiting, pruritus, respiratory depression and drowsiness and/or giddiness were elicited. Backache and inflammation at the site of insertion of the epidural catheter were also elicited in patients on regional analgesia. They were also asked if they were satisfied with the Pain Service. RESULTS: Out of 2,024 patients, 60% had undergone caesarean section and 40% had undergone major gynaecological procedures. Continuous intravenous morphine, the most common mode, was used in 83% of obstetric patients and 47.9% of gynaecological patients. Only one obstetric patients and one gynaecological patient complained of severe pain at rest while 12 obstetric patients and 3 gynaecological patients had severe pain on movement or coughing. Side-effects, most commonly nausea and/or vomiting and drowsiness and/or giddiness, were present in 8.8% of obstetric patients and 32.5% of gynaecological patients. 99.3% of obstetric patients and 99.6% of gynaecological patients were overall satisfied with their analgesia. CONCLUSION: Our post-operative analgesic modes were found to be effective and safe. PMID- 9407762 TI - Sickness absence in private sector establishments in Singapore. AB - AIM: A study was conducted in June 1995 to determine the current level of sickness absence in Singapore. METHOD: The questionnaire survey was part of a larger labour market survey conducted quarterly by the Ministry of Labour, and covered 3,553 private sector establishments employing 25 or more employees. RESULTS: Overall, 14.4% of the 628,477 employees took sick leave, while the percentage of working days lost due to sick leave, excluding maternity leave, was 1.1%. An average of 3.2 days of medical leave were taken per person per year. Industry specific characteristics seemed to have more influence on sickness absence than establishment size, employee's sex and occupation (viz, professional versus clerical versus production staff), number of hours worked and overtime work. Over 60% of the establishments, particularly larger companies and those in manufacturing, implemented measures to control sickness absence, most commonly counselling, disciplinary procedures and attendance allowance or bonus. Over 13% monitored sickness absence using computerised records. CONCLUSION: Comparing with overseas sickness absence (lost time) rates, the rates observed in this study do not appear high. PMID- 9407763 TI - Necrotizing fasciitis--an unusual presentation of miliary mycobacterium tuberculosis. AB - We report an immunocompromised patient who presented with necrotizing fasciitis as the initial presentation of miliary tuberculosis. The diagnosis of miliary tuberculosis was delayed resulting in prolonged morbidity and hospital stay. The lesson from this report is that tuberculosis should be recognised as an uncommon cause of necrotizing fasciitis in an immunocompromised patient, especially if the response to prompt and standard initial treatment is unsatisfactory. PMID- 9407764 TI - Carbimazole-induced agranulocytosis--a report of 2 recent cases. AB - Carbimazole is a useful antithyroid drug with a rare potentially fatal complication of agranulocytosis. We report 2 cases presenting with this problem. One was treated supportively with barrier nursing and broad spectrum antibiotics, and the other needed use of a haemopoietic growth factor, granulocyte colony stimulating factor (G-CSF). As it is indeed possible for thyrotoxic patients who developed agranulocytosis with carbimazole to have the same complication with propylthiouracil, once agranulocytosis had resolved, both patients were treated with radioiodine to maintain euthyroidism. Carbimazole-induced agranulocytosis usually spontaneously resolves within 1 to 2 weeks of stopping the drug. The use of haemopoietic growth factors to stimulate the proliferation and differentiation of progenitor cells, accelerates neutrophil recovery, as in our first case discussed. We recognise that agranulocytosis from carbimazole is a rare, life threatening complication. Instead of awaiting spontaneous recovery, the use of haemopoietic growth factors certainly seems a justifiable option, with a promise of a reduction in morbidity and mortality. PMID- 9407765 TI - Distal renal tubular acidosis and hereditary elliptocytosis in a single family. AB - Distal renal tubular acidosis (RTA) and hereditary elliptocytosis (HE) are apparently distinct, genetic conditions. We report a family with 3 children having both hereditary elliptocytosis and distal renal tubular acidosis. The simultaneous occurrence of these two conditions in three siblings could be due to covariations in the same family, although a possible contiguous gene syndrome for distal RTA and HE cannot be excluded. This report emphasises the importance of excluding a renal tubular defect in any child who presents with elliptocytosis and failure to thrive. PMID- 9407766 TI - Sulphite oxidase deficiency--a report of two siblings. AB - Isolated sulphite oxidase deficiency is a rare metabolic disorder characterised by neurological abnormalities, lens subluxation and seizures. Inheritance is autosomal recessive. We report two siblings with onset of clinical symptoms at 6 months of age, progressing to severe mental retardation, spasticity and seizures which were difficult to control. One of the siblings had lens subluxation. Diagnosis is made upon the increased levels of urinary sulphite, and high plasma S-sulphocysteine and thiosulphate levels. No treatment is known to be of help. Prenatal diagnosis is possible from the analysis of uncultured chorionic villus material for sulphite oxidase. PMID- 9407768 TI - What you need to know: fitness for work. PMID- 9407767 TI - Splenic lymphoma with villus lymphocytes--an uncommon cause for lymphocytosis. AB - We describe the clinical and laboratory features of four patients who presented with mild to moderate lymphocytosis but with no peripheral lymphadenopathy. These patients in the past, would have been classified as chronic lymphocytic leukaemia (CLL). However, it is now realised that chronic lymphoproliferative disorders are very heterogeneous and the clinical and laboratory features of our patients would support a diagnosis of splenic lymphoma with villus lymphocytes (SLVL) with characteristic morphological features. SLVL usually runs a benign clinical course but symptoms related a benign clinical course but symptoms related to splenomegaly or hypersplenism may be a problem. Splenectomy is considered the treatment of choice in these patients. Two of our patients had splenectomy and the other two patients are on regular follow-up without any specific treatment. It is therefore important to recognise this uncommon condition and also to differentiate it from CLL. PMID- 9407769 TI - Clinics in diagnostic imaging (29). Thalassaemia major with iron overload. AB - A 5-year-old girl presented with lethargy, anaemia and facial distortion. Both parents had beta-thalassaemia minor. Radiographs confirmed the characteristic features of thalassaemia major. A treatment regime comprising regular blood transfusions was commenced. The basis of the radiographic changes and the current role of magnetic resonance imaging, particularly with respect to assessing iron overload, are emphasized. PMID- 9407770 TI - Emotional effects on emergency room staff. PMID- 9407771 TI - Development of the 'Euro Rota' in A & E. AB - This paper outlines the basis for and terms of the European Working Time Directive, and its implementation in the Accident and Emergency (A & E) department of a UK hospital. The author describes how the directive was applied to the existing requirements of staffing the A & E department, including reaching a majority agreement on shift times. The new A & E staffing rota was implemented on 14 May 1997, and initial evaluation, after 6 weeks' running, is given. PMID- 9407772 TI - Grieving after a sudden death: the impact of the initial intervention. PMID- 9407773 TI - Children and the ingestion of alcohol: a statistical analysis of children attending an A & E department. AB - The results of a survey of children attending an Accident and Emergency (A & E) Department following ingestion of alcohol are reported. Data were analysed to indicate age and sex of children, date and time of attendance, method of transport to hospital, whether accompanied to hospital and if so by whom, and whether admitted or discharged. Trends in children's consumption of alcohol, as observed by the author, are discussed. PMID- 9407774 TI - Job interviews: tips and techniques. AB - This paper outlines appropriate preparation for a job interview, including preparing yourself to focus on your own personal assets and on what you can bring to the job. The various kinds of interview questions are examined: the traditional- 'tell me about yourself'; questions you dread because they will home in on 'weaknesses' and the unusual, open-ended questions intended to uncover specific information. Suggestions are given on how to use the experience of an interview to your own advantage, whether your application is successful or not. PMID- 9407775 TI - Becoming an A & E nurse. AB - This is the first of four papers which examine the work of Accident and Emergency (A & E) nurses. The descriptions in this and the following three papers have emerged from an ethnomethodological study which sought to obtain the views of what A & E nurses believed their work to be. All are rooted in nurses' accounts of the ways in which nursing work is talked about and accomplished within the A & E setting. It should be noted that all four papers describe the ordinary rather than the extraordinary and should hold no surprises for those familiar with A & E nursing work. This first paper explores the ways in which nurses become A & E nurses, however, before descriptions can be put forward, an introduction to the study from which they have emerged needs to be made. PMID- 9407776 TI - Visiting a Level I Trauma Centre in the USA. AB - A visit to a Level I Trauma Centre (Trauma Centre) in October 1996 is described. A group of seven emergency nurses from Hong Kong visited a major trauma facility in Minnesota USA, the Ramsey Medical Center, and took part in a Trauma Nursing Core Course (TNCC). The writer was one of the group and would like to thank everyone there who offered the group a warm reception and invaluable instruction. PMID- 9407778 TI - Phenomenon of sudden death: Part 2. PMID- 9407777 TI - Severe pre-eclampsia and eclampsia: a broad overview with discussion of the nursing care required for the eclamptic patient. AB - Severe pre-eclampsia and eclampsia are conditions which are rarely seen in A & E departments today. Although the percentage of patients suffering from these conditions is extremely small, the author has been involved in the care and treatment of two patients with this severe complication of pregnancy. A lack of clear guidelines on this subject was the motivation which prompted further study. This paper covers definitions of pre-eclampsia, eclampsia, history, incidence, risks and pathophysiology. The main discussion is the nursing care and treatment of the eclamptic patient in the acute phase of the condition. The author hopes that by the end of this paper it is evident that all health care professionals should be aware of this dangerous condition and know the appropriate treatment to give to ensure the best quality of care possible. PMID- 9407779 TI - Discharge planning from A & E: Part I. AB - The discharge of patients from hospital has always been a vital part of the nurse's role. The government recognized the need for health personnel to plan effective discharge of patients from hospital and guidelines were produced by the Department of Health in 1989. While these were aimed more at the inpatient than the Accident and Emergency (A & E) attender, many of the recommendations can be applied to the emergency setting. Nowhere more than A & E does this create a management problem for patients, carers and colleagues alike. Those patients that belong to vulnerable groups, the elderly, the homeless, children and the mentally ill require a comprehensive discharge programme, utilizing the skills and knowledge of a number of community care personnel. Current practices are explored and recommendations made for future management. Part 2 of the study, covering appropriate discharge advice for patients who do not need to reattend the department, will be published in 1998. PMID- 9407780 TI - Should relatives be invited to witness a resuscitation attempt? A review of the literature. AB - This review of the literature focuses upon the somewhat controversial question of whether relatives should be invited to witness the resuscitation of a family member. The pioneering study undertaken at the Foote Hospital, Michigan is explored, as is much of the anecdotal case study material available. The review focuses around three main areas: Allowing the relatives into the resuscitation room. Success of the encounter. The effects upon the relatives who witness the event. It is clear from this review that opinion about the presence of relatives in the resuscitation room remains inconclusive, with an apparently equal balance of both positive and negative family responses during the resuscitation attempt. However, whilst there is some evidence regarding short term effects on relatives who actually choose to witness a resuscitation attempt, there is little evidence to suggest what the long-term effects are likely to be. This is an aspect of resuscitation that certainly merits further exploration. PMID- 9407781 TI - The prisoner. PMID- 9407782 TI - A helping hand during a shattering time. PMID- 9407783 TI - The Florence Nightingale Foundation. PMID- 9407784 TI - Where do I begin? Writing for publication. AB - Nursing is changing. All nurses, clinical and management, are being encouraged to write. Some educators are under considerable pressure to write as part of their jobs. The university system has always insisted that teaching staff publish and many nursing colleges and schools are now joining the higher educational system. This article is about getting started in writing. It describes what to write, who to write it for and how to write for publication. PMID- 9407785 TI - The Infu-surg pressure infuser bag. PMID- 9407786 TI - Protecting your future. PMID- 9407787 TI - Evidence-based practice in perioperative nursing: a literature review and suggestions for change. AB - Evidence-based practice (nursing practice that relies on information generated from the results of scientific research) is widely recommended as the method of nursing practice of the future. This is particularly the method of choice in perioperative nursing. In this article the literature on evidence-based practice is reviewed with particular emphasis on the role of the perioperative nurse, research utilization, and barriers to evidence-based practice. Suggestions for change to promote evidence-based practice are also outlined. PMID- 9407788 TI - Orderlies and aides merging into O.R. attendants. AB - It was always my belief that perioperative nurses should be in the operating room performing nursing duties, not in the role of cleaning instruments and picking tables. New technology introduced over the past decade required longer schedules for nurses and the introduction of additional support staff. Because of the longer schedules, it became more cost effective to hire support staff to assist nurses in non-nursing functions. Recently, I was involved in a program which reorganized one aspect of our OR department and merged two existing support positions into one. Using ORNAC standards, CAS standards and CSS standards, I accepted the challenge of organizing the supplies of the department, merging orderlies and aides into one support group, training and documenting the important assignments given to this group and orienting staff to this new role. PMID- 9407789 TI - Cervical plexus block for carotid endarterectomy: a nursing care plan. AB - Historically, carotid surgery is identified in the operating room as a major surgical procedure. Although the surgical intervention remains the same, a regional anesthetic technique calls upon perioperative nurses to utilize their assessment and planning skills astutely preparing innovative nursing interventions that enable successful patient outcomes. The key to a successful nursing care plan for a carotid endarterectomy performed under cervical plexus block is an awareness of the patient's physiological needs as well as the environmental influences they may be experiencing. As the administration of regional anesthesia for major surgery become more prevalent, there is a resurgent demand for traditional holistic nursing interventions in the operating room. The perioperative nurse must couple technical expertise with intuitive assessment skills and administration of compassionate nursing care. PMID- 9407790 TI - The French patient. PMID- 9407791 TI - [Compliance with health regimens: statistical verification of conceptual structure]. AB - The use and definition of the concept compliance has been variable. The concepts compliance, adherence and co-operation have been used in the English literature. The terminology in the Finnish literature is also variable such concepts as hoitomyontyvyys, hoitomyotays, hoitokuuliaisuus and hoitoon sitoutuminen have been used. The purpose of this study is to illustrate the empirical structure of the concept compliance. The baseline was the definition of compliance of young diabetics. According to this definition, compliance is an active, intentional and responsible process by which young diabetics work to maintain their health in collaboration with the health care staff. The data were collected (1994) by a questionnaire from young diabetics aged 13-17 years (N = 403). To test the structure of the concept compliance, LISREL-analysis was used. The results of the study support the baseline definition of compliance. PMID- 9407792 TI - [From man to father--a review of nursing research]. AB - This literature review describes the work that has been done in the field of nursing research on growth into fatherhood starting from pregnancy and through the infant's first year of life. The purpose is to describe and analyse the existing research knowledge on this subject. The material of this review consists of 56 articles derived from the Cinahl (1983/1995) and Medline (1966-9/1995) databases using the keywords father, fathers, fathering and fatherhood. The method of content analysis was employed. The focal concern was with the contents of the articles and with the research designs and methods used. Five main content areas were identified: man's health, growth from manhood into fatherhood from the points of view stress and social support, fatherhood as a significant life experience, the relationship between father and child, the couple relationship and family functioning. It is obvious that more information is needed in Finland on male processes also in nursing research so that we can better understand those processes and further improve nursing interventions. Male processes have primarily been studied by using quantitative methods and cross-sectional settings. The main difficulties with the studies have been their small sample sizes and accordingly lack of generalizability. More qualitative information is required so that we can reach a deeper understanding on male processes, expectations and wishes. PMID- 9407793 TI - [Family caregivers' expectations of respite care of patients with dementia]. AB - The aim of this study was to describe the family caregivers' expectations of the respite care of demented patients. The data was collected using structured theme interviews. The participants (n = 15) of this study were family members of patients suffering from moderate or severe dementia. The family caregivers used respite care as a mode of support either regularly or occasionally. The data was analyzed using content analysis and classified in categories based on the theoretical frame of the study. The results imply that family caregivers expect information concerning dementia, its causes as well as care possibilities. Above all the family caregivers expressed the need to know about the existing support services: domestic help, day care or respite care, voluntary nursing services etc. Also the severe need for psychological support was revealed. Family caregivers wished to have more opportunities to discuss the oppressive matters with the nursing staff. Health professionals need to know more about the family members as well as their expectations and perceptions. That enables them to intervene more effectively to ease the burden of adjustment and facilitate the continuous involvement in care. PMID- 9407794 TI - [Patients' perception of surgical care. A study at HUCH, Surgical Hospital in 1994 and a comparison of study results from 1990]. AB - The purpose of this study was to assess patient satisfaction with care in HUCH, Surgical Hospital 1994 and to compare the results to the results of a-study done with the same questionnaire in 1990. The data were collected using SPRI:s (Sjukvardens och socialvardens planerings- och rationaliseringsinsitut) questionnaire from patients (N = 215) at seven wards. The response rate was 72. The structured questions were analysed by using the SOLO calculating program and the open-ended questions by using content analyses method. The results are presented in percentages and cross tabulations in the article. According to the results the patients were elder, suffered from more serious diseases and stayed for a longer time in the hospital than 1990. Patients' satisfaction was improved with information and nursing. However, even if the information was improved, most patients wanted more written information. On the whole, patients were satisfied with nursing care but some criticism was expressed concerning environment. PMID- 9407795 TI - [Nursing research in Finland from 1950s to the present]. AB - In Finland nursing education and research in universities was established in 1979. Research in nursing, however, was done in some amount even before in other disciplines and in the Institute of Nursing Research. In this article, nursing research during 1958-1995 is described, based on the articles in the Finnish Yearbook of Nursing (1958-1988), in the Journal of Nursing Science (Hoitotiede, 1989-1995) and academic licentiate and doctoral dissertations. The analysis indicates strong growth of nursing research in Finland since 1950s. The content of research is multidimensional. The nature of research is quantitative and descriptive; many of the doctoral dissertations in 1990s are qualitative. The research interest has been mainly the patient. Articles in research methods and concept analysis are rare. PMID- 9407796 TI - [Nursing research on the dynamics of hope in the elderly--a review]. AB - The dynamics of hope--the dialectics between hope and hopelessness--is a central dimension in the essence of human being, in health and illness, as well as in nursing. In this paper nursing research on hope and hopelessness in elderly people is described. CINAHL- and MEDLINE EXPRESS-database were used as sources of research material. The purposes, the target populations, the methods of data collection and the central results from 10 research articles were analyzed using the method of content analysis. In addition qualitative content analysis was used to illustrate the dynamics of hope. The research on the dynamics of hope in nursing science has mostly focused on young adults, and less on elderly people. Most of the studies on hope and hopelessness in elderly people were descriptive, cross-sectional research based on quantitative assumptions. The dynamics of hope was described as the dynamics of life and death in which elderly people's significant relationships, feelings and the ability to act were central elements. Qualitative, longitudinal designs are needed in nursing science in order to capture the essence of the dynamics of hope in elderly human beings. PMID- 9407797 TI - ["A good person does not feel envy"--envy in nursing communities]. AB - The purpose of this study was to describe the nature, manifestation, and amount of envy among the staff of a nursing community. The definition of envy is commonly based on views of essence of envy and organisational culture. The population study consisted of random sample of 120 subjects drawn from among the employees in the Kainuu Central Hospital. Frequency and percentage distributions were used to present the data. The correlations between the variables were examined using cross-tabulation. Summarized variables were formed for nature of envy, and the Cronbach alpha coefficient was used to test the internal consistency of those. Factor analysis and cross-tabulation were also used. Open ended questions were analysed by qualitative contact analysis. These results were used to complement quantitative data. The results of this study indicated that the employee's view of his/her official position in the nursing society, his/her relations with his/her fellow workers and the management as well as the relations to other nursing societies are all related to enviousness. The employees's view of his/her official position intensified his/her feelings of envy, if he/she had other negative feelings (anxiety, dissatisfaction with him/herself, and feeling if looks could kill). If the employee was ambitious and hard-working, his/her envy manifested in a comparison of his/her own work and the work of other employees. The major object of envy was fellow workers' salary. Envy was also caused by new, proficient, and senior co-workers and possible favourites or proteges of management. Envy towards other nursing community was generated by alleged differences in the amount of labour, or by the charge nurse's greater interest in other section. Employees coped with envy by hiding these feelings and being modest. Women coped with envy by being silent. PMID- 9407798 TI - Lee Ann Grogan. Interview by Kimberly Allen. PMID- 9407799 TI - Breast cancer legislation: a review of federal and state initiatives. PMID- 9407800 TI - BSN education. PMID- 9407801 TI - My perspective as an ADN student. PMID- 9407802 TI - Why I chose a diploma program. PMID- 9407803 TI - Perspectives of a man in nursing. PMID- 9407804 TI - Behind the scenes on the campaign trail. PMID- 9407805 TI - Tips on being a mom and a nursing student. PMID- 9407806 TI - 1996 Fuld Scholar winning essay. Advocating for ourselves: the emerging role of nurses as patient advocates within multidisciplinary health care teams. PMID- 9407807 TI - Euthanasia and assisted suicide. PMID- 9407808 TI - U.S. supreme court upholds bans on physician assisted suicide in New York and Washington. PMID- 9407810 TI - [Early return home--a challenge]. PMID- 9407809 TI - A lifetime's experiences in a day. PMID- 9407811 TI - [Changes in child care leave]. PMID- 9407812 TI - [Drug abuse. The child is the goal--and the means. Interview by Erk Dale]. PMID- 9407813 TI - [International health conference: the Jakarta Declaration on the health promotion initiative]. PMID- 9407814 TI - [Mothers with rheumatism: free them from the stress of nursing]. PMID- 9407815 TI - [A personal milk bank in the State Hospital]. PMID- 9407816 TI - [Molde: maternity department with everyday care. Interview by Eldri H. Fagerlund]. PMID- 9407817 TI - Trauma multidisciplinary QI project: evaluation of cervical spine clearance, collar selection, and skin care. AB - PURPOSE: To evaluate data regarding cervical collars and develop guidelines related to product selection, cervical spine clearance, and skin care. METHODS: Production selection and skin care were evaluated by multidisciplinary teams review of QI data. RESULTS: The multidisciplinary teams identified a c-spine clearance algorithm, a methodology for continuous skin evaluation, and a new policy for skin care under the c-collar. CONCLUSIONS: Focused attention to problems surrounding cervical spine immobilization can decrease patient complications. PMID- 9407818 TI - Neurotrauma family interventions. AB - TOPIC: Family responses to neurotrauma PURPOSE: To highlight the role of trauma nurses in providing education and support to families of neurotrauma patients SOURCES: Published literature and clinical experience CONCLUSIONS: Family centered care can provide many positive outcomes: e.g., reduced stress, decreased anxiety, decreased powerlessness, increased understanding and support. The success of the patient's rehabilitation often depends on the care and involvement of their families. PMID- 9407819 TI - Development of a policy for patients receiving neuromuscular blocking agents. AB - PURPOSE: To develop a policy to standardize the care of patients paralyzed with neuromuscular blocking agents (NMBAs). METHODS: Criteria were developed to assess adherence to the policy. Categorical and continuous data were collected and analyzed. RESULTS: Deficiencies were identified in frequency of train-of-four monitoring and in carrying out daily discontinuation of NMBAs to allow effects of paralysis to dissipate. CONCLUSIONS: The policy was useful as an educational tool. A NMBA order form should be developed to improve adherence to the policy. PMID- 9407821 TI - There's no place like home. PMID- 9407820 TI - The trauma coordinator's top strategies in the art of problem review (without a power base). PMID- 9407822 TI - The demand for evidence-based practice. PMID- 9407823 TI - Conference for prevention of incontinence. PMID- 9407824 TI - Stricture and pouchitis after ileal pouch-anal anastomosis. AB - Ileal pouch-anal anastomosis is a surgical procedure used for the treatment of people with chronic ulcerative colitis and familial adenomatous polyposis. The surgery is intended to preserve anal sphincter function, but it carries a risk for certain complications, including pouchitis and anastomotic stricture. The purpose of this article is to review the clinical manifestations, causes, and treatment of anastomotic stricture and pouchitis after ileal pouch-anal anastomosis. PMID- 9407825 TI - Improving stoma management in the low-vision patient. AB - Teaching the low-vision patient with an ostomy to manage a stoma independently provides a significant challenge for the WOC nurse. Although guide strips, lighted mirrors, and other handheld devices are available to patients with impaired eyesight, these products, when used alone, may be inadequate for the patient with an ostomy. In addition, there is a paucity of literature available to the WOC nurse describing specific interventions for the low-vision patient with an ostomy. Common visual problems encountered when managing people with an ostomy are reviewed. Specific interventions are discussed, including products designed to assist the low-vision patient with an ostomy with stoma care. A case study is provided that underscores the importance of consulting a low-vision specialist in the care of the patient with a new ostomy who has low vision as a result of albinism. PMID- 9407826 TI - Alternatives for the pharmacologic management of urge and stress urinary incontinence in the elderly. AB - Urinary incontinence in the elderly is a common but typically remediable condition that is frequently managed by pharmacotherapy. However, because of age related changes, the response to a specific dosage of a certain drug administered to an elderly person may be quite different than the response experienced by a younger individual. This article will review general principles of pharmacotherapy in the elderly patient and pharmacologic alternatives for the management of urge and stress urinary incontinence. PMID- 9407827 TI - The effect of bladder training on urinary incontinence in community-dwelling older women. AB - OBJECTIVE: The effect of bladder training on urinary incontinence among a group of community-based older women was investigated. DESIGN: Quasi-experimental. SETTING AND SUBJECTS: Nineteen functionally independent, community-dwelling women, aged 64 to 88 years, with a history of urinary incontinence occurring at least once a week, participated in the study. METHODS: Bladder training methods consisted of mandatory voiding schedules, self-monitored voiding records, and weekly telephone communication between subject and researcher. MAIN OUTCOME MEASURES: The number of incontinent episodes over a 1-week period was used to determine the effectiveness of bladder training. RESULTS: The mean frequency of incontinent episodes at baseline was compared to the frequency of incontinent episodes at both end of treatment and at 6-month follow-up. CONCLUSIONS: Bladder training is effective in reducing episodes of urinary incontinence in community dwelling older women. PMID- 9407828 TI - Patient with constipation requiring a bowel management program. PMID- 9407829 TI - Coping with wandering behavior. PMID- 9407830 TI - Mental illness in America. PMID- 9407831 TI - Speak into the computer, please. PMID- 9407832 TI - Can substance abuse among adolescents be predicted? PMID- 9407833 TI - Nutrition and mood. PMID- 9407834 TI - Selective serotonin reuptake inhibitors. PMID- 9407835 TI - Chronic school absence linked to psychiatric disorders. PMID- 9407836 TI - Mental health and cardiovascular disease. PMID- 9407837 TI - Anxiety disorders website launched. PMID- 9407838 TI - St. John's wort study launched. PMID- 9407839 TI - Post registration study: the way ahead. PMID- 9407840 TI - The missing links between practice and theory. PMID- 9407841 TI - How to get the credit for your previous learning. PMID- 9407842 TI - The social construction of medicine. PMID- 9407843 TI - The philosophy of conventional medicine. PMID- 9407844 TI - The re-emergence of complementary therapies. PMID- 9407845 TI - Presentation to a small group. PMID- 9407846 TI - Planning for learning. PMID- 9407847 TI - Overcoming parochialism: interdisciplinary training of the generalist team. AB - The work force that will staff most health care systems of the future will include a complex array of professionals working together in teams. The traditional inpatient model of patient care has been only multidisciplinary- nurses, medical social workers, dietitians, pharmacists, and physicians, all interested in each patient, but with divided responsibilities, training formats, and faculties--whereas interdisciplinary teams openly share decision making, expectations for care, goals for the team, and mutual respect. PMID- 9407848 TI - What women nursing students want & what nursing education owes them. AB - What do women want from nursing? Students have needs, motivations, and expectations related to their selection of a career. What are they? These questions were the subject of a research project in which women nursing students were given the opportunity to speak "in their own voices." The findings were both interesting and disturbing. While some of the results could be anticipated, others were unexpected. Only a brief summation of the research project will be presented here; a detailed description will be reported elsewhere. Instead, we will focus on what the findings reveal and what they imply for the nursing profession. PMID- 9407849 TI - Bodies of knowledge. AB - I learned the hard way, the old-fashioned way. I did my first pelvic exam on a colleague, who then did her first on me. We were terrified amateurs, willing to endure all just to get through. "You did great!" we congratulated each other, masking the inevitable discomfort with hearty words of encouragement. I did my second pelvic exam on a patient. What that patient experienced, I'll never know, because she didn't tell and I didn't ask. It took a year or two for me to begin to feel comfortable with the procedure and more years to feel skilled at it. And still I couldn't gauge my patient's experience. I'd say, casually, "You okay?" as I inserted the speculum. She'd say "Uh huh." Sometimes she convinced me, sometimes not. PMID- 9407850 TI - Rural health care & interdisciplinary education. AB - In response to the call for more rural health service providers and health education with an interdisciplinary focus, James Madison University's School of Health and Human Services initiated an undergraduate education program in rural Virginia. The project enhanced the attractiveness to students of rural health practice and exposed them to the effectiveness of collaboration among health professionals. PMID- 9407851 TI - For the health of Smith Island. AB - It is not much more than thirteen miles from Ewell to the hospital in Crisfield, but for residents of the small waterfront community, twelve and a half of those miles are across the open waters of the Chesapeake Bay. So it is not surprising that a program charged with increasing health care access for rural populations uses the island to introduce future practitioners to the needs of such underserved communities. Located roughly in the middle of the Chesapeake Bay, Smith Island falls within the boundaries of Somerset County, the poorest of Maryland's Eastern Shore counties. For the Eastern Shore Area Health Education Center (ESAHEC), a state-supported but private nonprofit agency founded to foster rural health care, Smith Island offered a perfect environment to expose an interdisciplinary group of health professions students to the realities of rural health care. Through a joint venture with the Somerset County Health Department, Smith Island is a destination for ESAHEC-sponsored trips that are part of the clinical experience for selected nursing, allied health, social work, and medical students in the University of Maryland system. PMID- 9407852 TI - A community crossword puzzle. An interdisciplinary approach to community-based learning. AB - Finding methods to involve communities in the education of health professionals presents a challenge to educators. An East Tennessee State University program found that a Community Crossword Puzzle helped students and faculty meet course objectives within the interdisciplinary curriculum of a rural health care education program. PMID- 9407853 TI - Taking aim at interdisciplinary education for continuous improvement in health care. AB - Over the last 30 years, nursing faculty have achieved varying levels of success in their efforts to engage in interdisciplinary education. To sharpen the focus, the Institute for Healthcare Improvement sponsored a national demonstration project in which nursing faculty from four universities participated. PMID- 9407854 TI - Reengineering the corporate culture of hospitals. AB - As we move closer to the turn of the century, the American health care system continues to provide management experts with a wealth of case studies describing the winners and losers in today's business environment. Winners in this cost conscious era of institutional reengineering, said Senge in his best-selling book The Fifth Discipline (1990), are those organizations that bind together around a common identity, a sense of destiny, and a shared commitment to reconciling the existing threats to their survival. Senge suggests that the organizations that are able to tap people's commitment and capacity to learn at all levels of the structure will overcome the type of maladaptation to business threats so commonly seen in corporate failures today. PMID- 9407855 TI - NLN & NLNAC stand together. AB - It is my understanding that there is still some confusion among our members and the public about: (1) the status of the National League for Nursing as an accrediting agency and (2) the relationship between the NLN and the National League for Nursing Accrediting Commission (NLNAC), the new entity that now handles all accrediting activities. It is of crucial importance that these matters are well understood. Therefore, I am taking this means to provide information (some of which will be repeated from previous communications) that I hope will clear up the confusion. PMID- 9407856 TI - Access to library and information services. PMID- 9407857 TI - Complementing the NHS. PMID- 9407858 TI - Restoring confidence. PMID- 9407860 TI - Future shock. PMID- 9407862 TI - Setting the pace for vascular services. PMID- 9407859 TI - Specialists only. PMID- 9407861 TI - Man trap. PMID- 9407863 TI - A change that is here to stay. PMID- 9407864 TI - Far away and hard to find. PMID- 9407865 TI - Qualified nurses losing out. PMID- 9407866 TI - Lobby for libraries. PMID- 9407867 TI - Open invitation. PMID- 9407868 TI - Incident reports in secure psychiatric units. AB - In 1995 Sir Louis Blom-Cooper observed a 'picture of increasing violence in British psychiatric hospitals in recent years, although the rate of serious violence is mercifully low' (Blom-Cooper et al 1995). In the following report, the author examines the incidence and management of violent incidents in three secure psychiatric units in the West Country. PMID- 9407869 TI - Non-compliance rights. AB - This paper evaluates the right of a pregnant woman to be supported in her choice not to comply with, or participate in, prescribed treatment and monitoring of impaired glucose tolerance. The debate involves ethical points including concepts of personhood, rights-based theory and justice. PMID- 9407870 TI - Homophobia and nursing care. AB - Homophobia, according to Blumenfield (1992) is both the belief that heterosexuality is or should be the only acceptable sexual orientation and the fear and hatred of those who are sexually attracted to those of the same sex. This definition forms the basis for this article, which explores whether living in a homophobic society affects the mental health of gay people and whether gay people are able to access appropriate services should they suffer from a mental illness. The article also examines whether nurses hold homophobic attitudes and if so, the extent to which these affect their work with mentally ill gay clients. PMID- 9407871 TI - The use of bed rails: principles of patient assessment. AB - Bed rails are often used to prevent patients from injuring themselves through falling, but they may not always have the desired effect. In this literature review, the authors demonstrate that patients must be assessed in relation to clearly defined risk factors to prevent indiscriminate use of bed rails. PMID- 9407873 TI - Mental health nursing. PMID- 9407872 TI - Childhood eczema: community care. PMID- 9407874 TI - Neurological observations-1 Glasgow Coma Scale. PMID- 9407875 TI - Abuse, spies and videotape. AB - Hospital video surveillance of parents has led to convictions for child abuse. But as Anne Gulland reports, concerns remain over both the reliability of the evidence collected and the ethics of nurses' involvement. PMID- 9407876 TI - Fighter pilots. PMID- 9407877 TI - The UKCC elections had a low turnout. PMID- 9407878 TI - Controlling interests. PMID- 9407880 TI - Dirty little secrets. PMID- 9407879 TI - Out of the blues. PMID- 9407881 TI - The dark side. PMID- 9407882 TI - Tale of the unexpected. PMID- 9407883 TI - The weekenderInterview by Janet Snell. PMID- 9407884 TI - How nurses can influence policy. PMID- 9407885 TI - Regulation in a changing climate. PMID- 9407886 TI - Developing a nurse-led aural care clinic. AB - This article describes how a nurse-led ear care clinic was developed at the North Staffordshire Hospital NHS Trust to reduce patients' waiting time. A nurse was trained under the supervision of two consultants and one registrar, and an assessment of the service after the first eight months showed that the clinic was successful and popular with patients. PMID- 9407887 TI - Determining the causes of fertility problems. AB - This article is the third in our series about fertility (the first two were in the September 3 and October 1 issues). It looks at the initial clinical investigations that patients have to undergo. The next article in the series, on December 3, will look at the indications for assisted conception. PMID- 9407888 TI - Water births: client choice versus legal implications. PMID- 9407889 TI - Sexual revolution. PMID- 9407890 TI - Learning disabilities. Pain signals. PMID- 9407892 TI - Epidemiology in infection control. PMID- 9407891 TI - Infection control. All fall down. PMID- 9407894 TI - Caring for neonates. PMID- 9407893 TI - Hepatitis C: a bloody business. PMID- 9407895 TI - Triumphant return in a new role. PMID- 9407896 TI - Protocols and guidelines for managing wounds. AB - The management of a chronic wound is aimed at symptom control and maintaining the individual's quality of life. Wound-care guidelines will promote a co-ordinated and systematic approach to wound management. The administration of wound-care products according to protocol can enhance the care of patients with complex chronic wounds. PMID- 9407897 TI - Nurses' confidence in caring for patients with alcohol-related problems. AB - General nurses lack confidence and experience in caring for patients with alcohol related problems. There is a need for better basic training and continuing education for nurses on alcohol issues. A specialist alcohol nurse could help support patients and staff. PMID- 9407898 TI - Drugs used for pain relief in palliative care. AB - Nurses have a vital role in assessing, monitoring and evaluating pain. Pain control should be based on a logical step-by-step approach. Morphine is not a panacea; adjuvant drugs are often required. While drug therapy is an important aspect of pain control, management must encompass an holistic approach. PMID- 9407900 TI - Psoriasis. PMID- 9407899 TI - Promoting health in older people with diabetes. AB - More than half of the UK diabetic population is aged over 65 years. Nurses have an important role in promoting the health and well-being of older people with diabetes. Management of diabetes aims to relieve symptoms and prevent or delay the progression of long-term complications. The degree of glycaemic control should be decided on an individual basis. PMID- 9407901 TI - Kawasaki disease. PMID- 9407902 TI - The safety of inhaled corticosteroids in children. AB - There are some concerns about the long-term effects of inhaled corticosteroids in children. Various steps can be taken to minimise the risks of treatment. The nurse has a crucial role in improving patients' and parents' understanding of asthma and its treatment. PMID- 9407903 TI - Patient-controlled analgesia devices. AB - PCA is associated with a reduced length of hospital stay and savings in nursing time and resources. Pre-operative patient education is required for the successful use of PCA. Regular patient and equipment monitoring is vital to ensure effective pain relief. PMID- 9407904 TI - Love, courage, and honor. PMID- 9407905 TI - Legacies of our presidents. PMID- 9407906 TI - Nineteenth century. Science and the values of duty, will, and power. PMID- 9407907 TI - A place at the table. Nurses' experience with the Rockefeller Foundation. PMID- 9407908 TI - American women in the 1920s. PMID- 9407909 TI - The depression years: problems and progress. PMID- 9407910 TI - The influence of economic forces on American nursing. Post World War II--1945 to 1950. PMID- 9407911 TI - Values in conflict. The 1950s. PMID- 9407912 TI - Thinking about patients. Nursing science in the 1950s. PMID- 9407913 TI - Mothers & pediatric nursing. Changing values 1880-1960. PMID- 9407914 TI - Reflections on the occasion of the 75th anniversary of Sigma Theta Tau. PMID- 9407915 TI - Nursing and the women's movement. The legacy of the 1960s. PMID- 9407916 TI - Values that drove nursing science in the 1970s. PMID- 9407917 TI - Image & ideas. PMID- 9407918 TI - What values drive nursing science? An intellectual and social history of nursing- 1980-1997. PMID- 9407919 TI - Professional values and nursing mentorship, Australia. PMID- 9407920 TI - A journey of ideas. Taiwan. PMID- 9407921 TI - A heritage of healing. Reflections on nursing in Canada. PMID- 9407922 TI - The current status of nursing science in South Korea. PMID- 9407923 TI - The future, virtue-ethics, and Sigma Theta Tau. PMID- 9407924 TI - [Students' work is important]. PMID- 9407925 TI - [Continuous care chain]. AB - Experiences of a liason nurse work between a hospital and home health care in the Central health centre of Helsinki. Home health care consists almost solely of nursing of elderly people. Today it issues new challenges concerning how the services can meet the elderly people's needs and expectations as good as possible and how these services are coordinated to other social and health services for the elderly. The care of elderly people is focused too much on institutions, but an effort is made to change this by a service structure change. Its aim is to decrease the amount of hospital beds for chronic patients by simultaneously increasing out-patient care. The liason nurse pilot programme is aimed at supporting this structural change by developing discharge of patients and by simultaneously creating more individual and good-quality home health care services. PMID- 9407926 TI - [A night-shift service in the city of Helsinki]. AB - This activity is a part of care at patients' homes. It makes it possible that even seriously ill people can be taken care of at home more often than before. The patients can live at their homes as long as possible. According to the view of the society this activity has been more economic than hospital care. The service has been very popular and it's availability will be increased. The night patrols work every day from 8.30 P.M. to 6.30 A.M. There are two patrols every night and they consist of a nurse and a practical nurse or a psychiatric nurse. The patrols take care of about 40-50 patients per night. Over 60% of them are over 80 years old. Generally the aged patients with several illnesses and terminal care patients need care at night, but also for example accident, rheumatic and MS patients may need the service. PMID- 9407927 TI - [Hospital care at home]. PMID- 9407928 TI - [Home care fees and appropriate use of services]. PMID- 9407929 TI - [Esthetics in nursing]. AB - According to the author's opinion aesthetics in nursing is interaction with people and environment. Balance between respect of a human being and presence of the form, colour, rhythm and peace of environment is related to it. The aim of aesthetics is to bring pleasure, joy, comfort, hope and beauty and in addition to all this, to strengthen self-experience of a human being. Aesthetic atmosphere at its best can encourage a human being to use images which is psychic activity that gives deeper understanding concerning her events of life and their personal meaning. Aesthetics can promote health of a human being and improve the quality of life--when one thinks of patients and personnel. PMID- 9407930 TI - [An operations model for treating diseases of fat metabolism]. AB - This article illustrates an operations model for treating diseases of fat metabolism. Implementation of the model was began in the autumn of 1996 by Vakka Suomen sairaala in Uusikaupunki and by health centres in its region. Development of multiprofessional co-operation in screening and treating these diseases is a part of wider quality project work in treatment of cardiovascular diseases. The aim of this project is to clarify and unite research and treatment practices by creating a good common guide for all professional groups. A regional operations model concerning screening, diagnose and treatment will be planned. Cornerstones of drug-free treatment are diet, physical exercise and losing weight. Medication will be initiated if the goal is not achieved by the drug-free treatment. The model includes exact directions for initiation and follow-up of medication. Local education will motivate all health care professionals to follow the common principles. PMID- 9407931 TI - [Individual guidance model in smoking cessation]. AB - Smoking is the cause of many diseases and one of the most serious risk factors in our country. Despite of this fact smoking habits of clients visiting during surgery hours are only occasionally taken into consideration and documented in the patient records. The Finnish Heart Association started a project in spring 1996 which aims at reducing smoking of patients with coronary artery disease. The aim is to create an individual guidance model for giving up smoking--the model will be a part of daily nursing practice. The model is based on a transition phase model which is now applied for the first time in the Finnish health care. The core of the model is that when a smoker visits during surgery hours his preparedness to give up smoking is defined and guidance is directed according to that level. Also smokers who do not wish to give up smoking are included in the guidance. The guidance model is currently tested at the hospitals, health centres and occupational health care in the area of the towns of Imatra and Lappeenranta. Personnel of test sites was trained in using the model before the intervention was implemented. The intervention ends in spring 1998 and the first results of applicability and effectiveness of the model are received. PMID- 9407932 TI - Cervical cancer. AB - OBJECTIVE: To provide physicians and the general public with a responsible assessment of current screening, prevention, and treatment approaches to cervical cancer. PARTICIPANTS: A non-Federal, nonadvocate, 13-member panel representing the fields of obstetrics and gynecology, gynecologic oncology, radiation oncology, and epidemiology. In addition, 28 experts in obstetrics and gynecology, gynecologic oncology, radiation oncology, gynecologic surgery, and psychology presented data to the panel and a conference audience of 500. EVIDENCE: The literature was searched through Medline and an extensive bibliography of references was provided to the panel and the conference audience. Experts prepared abstracts with relevant citations from the literature. Scientific evidence was given precedence over clinical anecdotal experience. CONSENSUS PROCESS: The panel, answering predefined questions, developed its conclusions based on the scientific evidence presented in open forum and the scientific literature. The panel composed a draft statement that was read in its entirety and circulated to the experts and the audience for comment. Thereafter, the panel resolved conflicting recommendations and released a revised statement at the end of the conference. The panel finalized the revisions within a few weeks after the conference. CONCLUSIONS: Carcinoma of the cervix is causally related to infection with the human papillomavirus (HPV). Reducing the rate of HPV infection by changes in sexual behaviors in young people and/or through the development of an effective HPV vaccine would reduce the incidence of this disease. Pap smear screening remains the best available method of reducing the incidence and mortality of invasive cervical cancer. Persons with stage IA1 disease have a high cure rate with either simple hysterectomy or, where fertility preservation is an issue, by cone biopsy with clear margins. For patients with other stage I and stage IIA disease, radical surgery and radiation are equally effective treatments. These patients should be carefully selected to receive one treatment or the other but not both, as their combined use substantially increases the cost and morbidity of treatment. Women with more advanced, nonmetastatic disease should be treated with radiation. Recurrent cervical cancer confined to the pelvis should be treated with the modality not previously received. Radiation is recommended to palliate symptoms in patients with metastatic disease. PMID- 9407933 TI - Molecular genetics and pathogenesis of rhabdomyosarcoma. PMID- 9407934 TI - Genetics of pediatric central nervous system tumors. AB - PURPOSE: Pediatric central nervous system (CNS) tumors comprise a wide variety of histologic subtypes ranging from the benign juvenile pilocytic astrocytoma to the highly aggressive atypical teratoid/rhabdoid tumor. Although some brain tumors are seen in association with inherited genetic disorders which predispose to malignancies, most are sporadic. Current knowledge regarding the cytogenetic and molecular genetic events which have been implicated in the development or progression of common brain tumors in children in the subject of this review. METHODS: Combined cytogenetic and molecular genetic approaches, including fluorescence in situ hybridization, have been used to identify genomic alterations in different histologic types of pediatric brain tumors. RESULTS: The most frequent abnormality in primitive neuroectodermal tumor/medulloblastoma is an i(17q), present in approximately 50% of cases. This finding implicates the presence of a tumor suppressor gene on 17p, which is important in tumor development. A number of genes on 17p have been eliminated as candidates for this locus, including TP53. A tumor suppressor gene in chromosome band 22q11.2 has been hypothesized to play a role in atypical teratoid/rhabdoid tumors, and positional cloning strategies are in progress to identify a rhabdoid tumor gene. Chromosome 22 deletions are also seen in meningiomas and a small percentage of ependymomas, but it is not yet known whether the same gene is responsible for more than one malignancy. With regard to childhood astrocytomas, tumor-associated genetic changes have not yet been identified for the common juvenile pilocytic or low grade diffuse astrocytoma. In contrast, malignant anaplastic astrocytomas and glioblastoma multiforme have abnormalities similar to those seen in adults, including loss of alleles on 17p13 and TP53 mutations, trisomy 7, EGFR rearrangements, and loss of chromosomes 10 and 22. CONCLUSIONS: The presence of tumor-associated genetic abnormalities has clinical utility in a differential diagnostic setting, and has lead to the identification of genes which contribute to tumorigenesis. PMID- 9407935 TI - Hodgkin's disease in childhood: a review. PMID- 9407936 TI - Potential prognostic value of C-erbB-2 expression in medulloblastomas in very young children. AB - PURPOSE: The expression of the c-erbB-2 oncogene was studied in childhood medulloblastoma to evaluate its prognostic value, which has been claimed previously. PATIENTS AND METHODS: Tumor material from 45 patients < 15 years old at diagnosis was studied using 3 monoclonal antibodies against the internal and external domains of the c-erbB-2 oncogene product. RESULTS: Six of the 45 (13%) tumor specimens were found to be positive. C-erbB-2 expression was found more often in patients < 3 years old at diagnosis (4 of 15 patients, 27%) than in older patients (2 of 30, 6.6%). During the follow-up period (5.8 +/- 2.8 years) all patients with c-erbB-2 expression died of disease (after 1.2 +/- 0.7 years). Kaplan-Meier estimation revealed a highly significant correlation of c-erbB-2 expression and survival (p = 0.002). A further study of the expression of synaptophysin and the glial fibrillary acidic protein (GFAP) in the 45 tumors revealed a negative correlation of the expression of c-erbB-2 and these proteins. CONCLUSION: C-erbB-2, which may be predominantly expressed by less differentiated tumors, was found to delineate a poorer prognostic subgroup, especially when diagnosed in patients < 3 years old. PMID- 9407938 TI - The effect of ABO and Rh blood type on the response to intravenous immune globulin (IVIG) in children with immune thrombocytopenic purpura (ITP). AB - PURPOSE: Immune thrombocytopenic purpura (ITP) is a common childhood illness characterized by thrombocytopenia secondary to shortened platelet survival. Medical therapy includes corticosteroids, intravenous immune globulin (IVIG), and IV Rho (D) immunoglobulin (anti-D). Individuals with Rh-negative blood generally do not respond to treatment with anti-D, but little information is currently available regarding the potential relationship between blood type and response to IVIG. This study was designed to characterize the relationship between ABO and Rh blood type and the response to IVIG in children and adolescents with newly diagnosed ITP. PATIENTS AND METHODS: A retrospective chart review was performed for 52 children and adolescents with newly diagnosed ITP initially treated with IVIG by the Division of Pediatric Hematology-Oncology at Robert Wood Johnson University Hospital in New Brunswick, New Jersey. RESULTS: There were no significant differences in response rate or clinical outcome by ABO blood group or Rh type in children with ITP who received IVIG monotherapy as their initial treatment. CONCLUSIONS: ABO blood group and Rh type do not appear to be prognostic factors when IVIG monotherapy is the initial treatment for childhood ITP. PMID- 9407937 TI - Childhood B lineage acute lymphoblastic leukemia clonality study by the polymerase chain reaction. AB - PURPOSE: B cell precursors acute lymphoblastic leukemia (ALL) present rearrangements in the heavy chain immunoglobulin and T cell receptor genes, especially in the complementarity determining region 3 (CDR-3) and T cell receptor delta (TCR delta) (V delta 2 D delta 3) regions. These rearrangements may be amplified by the polymerase chain reaction (PCR) and used as clonal markers of B lineage ALL. Our purpose was to study clonality at the DNA level by PCR in B lineage ALL. PATIENTS AND METHODS: Fifty-three pediatric patients (36 with B lineage ALL, 7 with ALL-T, and 10 with nonlymphocytic disease) were investigated using consensus primers for the CDR-3 regions of IgH and TCR delta. RESULTS: Clonality was detected in 86.1% of the patients with B lineage ALL when the primers for the CDR-3 regions were used, in 41.6% when the primers for TCR delta were used, and in 91.6% when the two primers were used together. Biclonality was found in 22.5% and 6.6% of patients that have shown clonality for CDR-3 and TCR delta, respectively. Clonality was not detected in any other samples using these primers. CONCLUSIONS: PCR using CDR-3 and TCR delta primers can be used as an aid for B lineage ALL diagnosis and clonal evolution of theses disease. PMID- 9407939 TI - Pulsed high-dose dexamethasone therapy in children with chronic idiopathic thrombocytopenic purpura. AB - PURPOSE: The effectiveness of pulsed high-dose oral dexamethasone therapy in children with refractory chronic idiopathic thrombocytopenic purpura (ITP) is evaluated. PATIENTS AND METHODS: Seven children (5 to 16 years old) who were refractory to 2 to 5 conventional standard therapies were included in the study. Dexamethasone was administered orally at a dosage of 40 mg/m2 per day (maximum 40 mg/day) for 4 consecutive days as a cycle. The cycle was repeated once a month for 6 months. RESULTS: One month after the first cycle, partial responses of platelet counts (> or = 50 x 10(9)/L and < 150 x 10(9)/L) were observed in three patients (43%). At the end of the sixth cycle, two patients (29%) had complete responses (> 150 x 10(9)/L) and one had a partial response. However, only one patient (14%) remained partially responsive 1 year after completion of therapy. CONCLUSIONS: In contrast to what was observed in adults, this preliminary study suggests that pulsed high-dose oral dexamethasone therapy was not uniformly effective in children with chronic ITP. PMID- 9407940 TI - Accumulation of methotrexate in systemic tissues after intrathecal administration. AB - PURPOSE: To determine systemic concentrations of methotrexate (MTX) after intrathecal administration in a patient with active leptomeningeal cancer. METHODS: Tissues were obtained at autopsy, as requested by the patient, for MTX analysis after 5 doses of the drug were injected into his cerebrospinal fluid (CSF) 6 to 21 days before death. A radioligand assay was used to measure the concentration of MTX in multiple tissues. RESULTS: Outside the neuraxis, MTX was found in highest concentration in the liver, kidney, lymph nodes, and spleen. Lung and bone also had substantial concentrations. Skeletal muscle had the lowest concentrations of measurable drug and the drug was not detected in the heart. In retrospect, the patient's second course of systemic chemotherapy resulted in greater systemic toxicities because intrathecal MTX was given in conjunction with this course of treatment. In addition, hypersensitivity to intrathecal morphine may have been caused by concomitant intrathecal MTX therapy. CONCLUSIONS: Physicians administering intrathecal MTX must be aware of the paradoxically greater systemic exposure of the drug after intrathecal administration than after systemic administration, the need to expect greater systemic toxicities of intrathecal MTX, and the possibility that it may induce a hypersensitivity to concomitant intrathecal morphine. PMID- 9407941 TI - Immunosuppressive therapy vs. bone marrow transplant for severe aplastic anemia. PMID- 9407942 TI - Gene therapy with B7.1 and GM-CSF vaccines in a murine AML model. AB - PURPOSE: Characterization of B7.1 and GM-CSF vaccines on the induction of anti tumor immunity in a murine AML model. MATERIALS AND METHODS: Primary AML cells were retrovirally transduced with the murine costimulatory molecule B7.1, a natural ligand for the T-cell receptors CD28 and CTLA-4, or the cytokine GM-CSF. Mice were vaccinated with irradiated AML cells expressing B7.1 or GM-CSF before or after inoculation of wild type AML cells. RESULTS: Intravenous injection of irradiated B7.1 or GM-CSF expressing AML cells can provide long lasting systemic immunity against a subsequent challenge of wild type AML cells. Vaccination with irradiated B7.1 or GM-CSF expressing AML cells results in rejection of established leukemia when the vaccination occurs in the early stages of the disease. However, when the vaccines are administered > 2 weeks after leukemic inoculation, only mice which receive the GM-CSF vaccine are cured of leukemia. CONCLUSIONS: These results suggest that tumor burden and vaccine efficiency are most likely to be the limiting factors in the curative potential of tumor vaccines. Novel approaches such as this experiment could provide improved therapeutic outcomes in patients with AML and other cancers. PMID- 9407943 TI - Regulation of apoptosis by integrin receptors. PMID- 9407944 TI - SCID due to ZAP-70 deficiency. PMID- 9407945 TI - Role of the NF1 gene in leukemogenesis and myeloid growth control. PMID- 9407946 TI - Promoting apoptosis: a novel activity associated with the cyclin-dependent kinase inhibitor p27. AB - p27Kip1, a cyclin-dependent kinase inhibitor, is recognized as a negative regulator of the cell cycle. In this paper, we report that overexpression of p27Kip1 triggers apoptosis in several different human cancer cell lines. Using a recombinant adenoviral vector that expresses p27Kip1 (Adp27), we found that overexpression of p27Kip1 in MDA-MB-231 breast cancer cells induces apoptosis that was seen by a number of different techniques, including flow cytometry and in situ terminal deoxynucleotidyl transferase-mediated nick end labeling, flow cytometric assay for sub-G1 population, and 4',6-diamindino-2-phenylindole staining. Cleavage of poly(ADP-ribose) polymerase and degradation of cyclin B1, events that are known to be associated with apoptosis, were also observed following overexpression of p27Kip1. This is the first report indicating a role for p27Kip1 in induction of apoptosis. PMID- 9407947 TI - Mutations of the MEN1 tumor suppressor gene in pituitary tumors. AB - Although pituitary adenomas are monoclonal proliferations, somatic mutations involving genes that govern cell proliferation or hormone production have been difficult to identify. The genetic etiology of most pituitary tumors, therefore, remains unknown. Pituitary adenomas can develop sporadically or as a part of multiple endocrine neoplasia type 1 (MEN1). Recently, the gene responsible for MEN1 was cloned. To elucidate the potential etiological role of the MEN1 gene in pituitary tumorigenesis, 39 sporadic pituitary adenomas from 38 patients and 1 pituitary adenoma from a familial MEN1 patient were examined for MEN1 gene mutations and allelic deletions. Four of 39 sporadic pituitary adenomas showed a deletion of one copy of the MEN1 gene, and a specific MEN1 gene mutation in the remaining gene copy was detected in 2 of these tumors. The corresponding germ line sequence was normal in all sporadic cases. A specific MEN1 mutation was detected in a pituitary adenoma and corresponding germ-line DNA in a patient with familial MEN1. An allelic deletion of the remaining copy of the MEN1 gene was also found in the patient's tumor. Genetic alterations of the MEN1 gene represent a candidate pathogenetic mechanism of pituitary tumorigenesis. The data suggest that somatic MEN1 gene mutations and deletions play a causative role in the development of a subgroup of sporadic pituitary adenomas. PMID- 9407948 TI - TCL1 is overexpressed in patients affected by adult T-cell leukemias. AB - Among mature postthymic T-cell leukemias, adult T-cell leukemia (ATL) has characteristic clinicopathological entities. The association with the human T cell leukemia/lymphotropic virus type I is one of the distinctive etiopathogenetic features of this disease. However, unlike other acute transforming retroviruses, the human T-cell leukemia/lymphotropic virus type I lacks an oncogene within its genome. Other human postthymic leukemias, such as T prolymphocytic leukemias, involve mostly the CD4 cellular subset and share many similarities to ATLs (aggressive course, cutaneous involvement, CD4+, CD29+, CD45RA- phenotype, and alpha-naphthyl-acetate esterase positivity). A chromosomal rearrangement at 14q32.1, involved in translocations or inversions with either the alpha/delta locus [t(14;14)(q11;q32.1), inv14(q11;q32.1)], or the beta-chain locus of the T-cell receptor [t(7;14)(q35;q32.1)] is found. These rearrangements disregulate a gene, TCL1, located at the 14q32.1 region, that we show is physiologically expressed in CD4/CD8 double-negative thymocyte cells, but not in more differentiated CD4+ and CD8+ subpopulations. Here, using molecular and immunocytochemical analysis, we report that TCL1 is also overexpressed in 10 of 10 ATL specimens, indicating that this gene may play an important role in the pathogenesis of this disease. PMID- 9407949 TI - The DNA repair activity of human redox/repair protein APE/Ref-1 is inactivated by phosphorylation. AB - The human DNA repair protein apurinic/apyrimidinic endonuclease (APE) is a dual function protein that has important roles in both the repair of baseless sites that arise in DNA and in regulating the redox state of a number of proteins (Ref 1). Although previous attention has been focused on how the human APE/Ref-1 gene may be regulated at the DNA level, we have instead examined if APE/Ref-1 is phosphorylated, and if so how it may affect DNA repair activity. We demonstrate here that APE/Ref-1 is indeed a substrate for phosphorylation by the serine/threonine casein kinases (CK) I and II and protein kinase C. Notably, although phosphorylation by CKI and protein kinase C had no effect whatsoever on the ability of APE/Ref-1 to act at abasic sites in DNA, phosphorylation by CKII completely abolished DNA repair activity. That phosphorylation was responsible for the loss of abasic repair activity was concluded from experiments showing that inactive APE/Ref-1 could be reversed to an active DNA repair protein with phosphatase treatment. These results may help to explain the mechanism by which APE/Ref-1 switches from one unrelated function to another. PMID- 9407950 TI - The 95-kilodalton membrane glycoprotein overexpressed in novel multidrug resistant breast cancer cells is NCA, the nonspecific cross-reacting antigen of carcinoembryonic antigen. AB - Human breast carcinoma MCF-7/AdrVp cells display a novel multidrug resistance phenotype that is characterized by the overexpression of a 95-kDa membrane glycoprotein (p95) and by marked reduction in intracellular anthracycline accumulation, without overexpression of P-glycoprotein or the multidrug resistance protein MRP. p95 is also highly expressed in multidrug-resistant NCI H1688 cells derived from a human small cell lung carcinoma. Deglycoslyated p95 from NCI-H1688 cells was isolated by two-dimensional gel electrophoresis and then digested with trypsin. The tryptic peptides were analyzed by mass spectrometry and microsequencing. These analyses identified p95 to be identical to NCA-90, the nonspecific cross-reacting antigen related to the carcinoembryonic antigen (CEA). Further confirmation that p95 is indeed NCA-90 was obtained by Northern and Western blot studies using probes or antibodies specific for p95, NCA-90, or CEA family members. Western blot studies also revealed that CEA itself is overexpressed in MCF-7/AdrVp cells compared to parental MCF-7/W cells. The enforced expression of NCA-90 protein in HeLa cells stably transfected with NCA 90 cDNA did not result in increased resistance of the transfected cells to daunorubicin or a decrease in daunorubicin accumulation in the transfected cells compared to cells transfected only with the expression vector. However, a recent report by H. Kawaharata et al. (Int. J. Cancer, 72: 377-382, 1997) of diminished accumulation, retention, and cytotoxicity of doxorubicin in EJNIH3T3 cells in which enforced expression of CEA was accomplished leaves open the possibility that the overexpression of CEA, possibly in combination with that of NCA-90, could account at least in part for the drug resistant phenotype displayed by MCF 7/AdrVp cells. PMID- 9407951 TI - Overexpression of phospholipase C-gamma1 in rat 3Y1 fibroblast cells leads to malignant transformation. AB - Phospholipase C-gamma1 (PLC-gamma1) mediates signals from various extracellular origins to evoke cellular events such as mitogenesis. Previously, we reported that PLC-gamma1 was highly expressed in colorectal cancer and familial adenomatous polyposis, suggesting that PLC-gamma1 might be oncogenic. In this study, we have established rat 3Y1 fibroblasts that overexpress whole PLC-gamma1 and src homology 2 (SH2)-SH2-SH3 domain of PLC-gamma1. These cells showed a transformed phenotype and were tumorigenic when transplanted into nude mice. These results indicate that overexpression of PLC-gamma1 could transform rat fibroblasts, and the transformation is mediated by SH2-SH2-SH3 domain of PLC gamma1. PMID- 9407952 TI - Genome-wide search for loss of heterozygosity shows extensive genetic diversity of human breast carcinomas. AB - Deletions of genomic regions involving tumor suppressor genes are thought to be important in the initiation and progression of breast cancer. We conducted a genome-wide search for deleted regions in a series of 75 human breast carcinomas by studying the allelic patterns of 184 microsatellite markers distributed over all chromosomes and looking for loss of heterozygosity (LOH). We identified 56 regions of consistent LOH. Strikingly, every tumor had a different set of deletions. To study this complexity, we applied a phylogenetic-like type of analysis. Each region was involved in a certain proportion of tumors, ranging from 20 to 62%; the most frequently involved regions were on chromosome arms 8p, 11q, 16q, and 17p. There was a correlation (P = 0.005) between the level of LOH and the size of the tumors. Tumors with a high level of LOH were also highly proliferative and had a high mitotic index. PMID- 9407953 TI - A canalicular multispecific organic anion transporter (cMOAT) antisense cDNA enhances drug sensitivity in human hepatic cancer cells. AB - The human cMOAT gene encodes a membrane protein involved in the ATP-dependent transport of hydrophobic compounds. To determine whether cMOAT is associated with drug sensitivity, we transfected an expression vector containing cMOAT antisense cDNA into the HepG2 human hepatic cancer cell line. We observed a reduction in cMOAT protein, as well as an enhanced level of glutathione, in the antisense transfectants. The transfectants displayed an increased sensitivity to cisplatin, vincristine, doxorubicin, and the camptothecin derivatives, (4S)-4,11-diethyl-4 hydroxy-9-[(4-piperidinopiperidino)carbonyl oxy]dione hydrochloride triethydrate and 7-ethyl-10-hydroxycamptothecin, but not to etoposide, 3-[4-amino-2-methyl-5 pyrimidinyl)methyl]-1-(2-chloroethyl)-1-nitrosoure a, 5-fluorouracil, and mitomycin C. Results suggest that cMOAT levels are inversely correlated with those of glutathione, and that cMOAT and its related genes may be involved in the sensitivity of cells to certain anticancer agents. PMID- 9407954 TI - The I1307K APC mutation does not predispose to colorectal cancer in Jewish Ashkenazi breast and breast-ovarian cancer kindreds. AB - An increased incidence of colorectal cancer has been observed in breast and breast-ovarian cancer syndrome families, including those of Ashkenazi origin. Recently, a germ-line missense mutation in the APC gene, I1307K, was identified that may indirectly cause colorectal cancer in Ashkenazi Jews. To determine whether the excess of colon cancer in some breast-ovarian cancer families is related to the I1307K mutation, we evaluated 264 Ashkenazi Jews from 158 families. Most of these individuals had either a personal or a family history of breast and/or ovarian cancer, and 19.3% (51 of 264) carried one of the recurrent BRCA1 (185delAG or 5382 insC) or BRCA2 (6174delT) mutations. We detected the APC I1307K mutation in 7% (11 of 158) of the Ashkenazi Jewish families and in 4.5% (12 of 264) of the individuals participating in these studies. Of the families studied, 26.6% (42 of 158) had at least one case of colorectal cancer in a first , second-, or third-degree relative of the proband. Significantly, of the 12 individuals who possessed the I1307K mutation, none was diagnosed with colorectal cancer and none had a known first-, second-, or third-degree relative diagnosed with colon cancer. The results suggest that factors other than the I1307K mutation contribute to the increased incidence of colon cancer in Ashkenazi breast-ovarian cancer families. Our results emphasize that only a subset of Ashkenazi Jewish individuals with a family history of colorectal cancer should be viewed as candidates for genetic susceptibility testing for the I1307K APC mutation. PMID- 9407955 TI - Nuclear location and cell cycle regulation of the BRCA2 protein. AB - Women carrying a germ-line mutation in the BRCA1 or BRCA2 genes have a high risk of developing breast cancer, and loss of the wild-type allele in tumors suggests that these genes function as tumor suppressor genes. The BRCA2 gene encodes a 3418-amino acid protein with no significant sequence similarity to any known protein. To begin to elucidate the cellular role of BRCA2, we have raised antibodies to the BRCA2 protein and used these to study its subcellular localization and expression. We show that BRCA2 is a nuclear protein expressed in response to cell proliferation and that BRCA2 expression is initiated before DNA synthesis. PMID- 9407956 TI - Chemoprevention of pulmonary carcinogenesis by aerosolized budesonide in female A/J mice. AB - This investigation is part of a continuing effort to develop effective chemoprevention for carcinogenesis of the lung. The present study explores the use of aerosol administrations for this purpose. The agent selected for initial study was the synthetic glucocorticoid budesonide. This selection was based on previous work in which budesonide added to the diet was found to inhibit pulmonary adenoma formation in female A/J mice. However, high dose levels were required, i.e., of the order of 300 microg/kg, of body weight [L. W. Wattenberg and R. D. Estensen, Carcinogenesis (Lond.), 18: 2015-2017, 1997]. For aerosol administration of budesonide, a nose-only technique has been developed that entails nebulization of the compound dissolved in ethanol and subsequent stripping off of the solvent (less than 3 microl ethanol/liter of air remaining at the site of inhalation). The budesonide particles produced by the apparatus had a mass median aerodynamic diameter of less than 1 microm. An experiment has been carried out in which the inhibitory effects of aerosolized budesonide, given for 1 min six times a week, were studied. Concentrations of budesonide of 26, 81, and 148 microg/liter of air (calculated doses of 23, 72, and 126 microg/kg of body weight) were used. The aerosols were started 1 week after three oral administrations of benzo(a)pyrene (2 mg/20 g of body weight) to female A/J mice. All three doses of budesonide resulted in more than 80% inhibition of pulmonary tumor formation compared to the aerosol control and 90% or greater compared to mice not exposed to aerosol. The difference in inhibition is due to the aerosol procedure itself, which produces a reduction in tumor formation. A decrease in splenic weight (evidence of a systemic effect) occurred at all doses of budesonide. To the best of our knowledge, this is the first published effort at the use of aerosol administration to prevent neoplasia of the respiratory tract. The results of the present study show that administration of a potential chemopreventive agent by aerosol at a low dose can inhibit the occurrence of pulmonary carcinogenesis in female A/J mice. PMID- 9407957 TI - An association between the allele coding for a low activity variant of catechol-O methyltransferase and the risk for breast cancer. AB - Mounting evidence suggests that catechol metabolites of estradiol may contribute to the development of estrogen-induced cancers. O-Methylation, catalyzed by catechol-O-methyltransferase (COMT), inactivates catechol estrogens. COMT is polymorphic in the human population, with 25% of Caucasians being homozygous for a low activity allele of the enzyme (COMT(LL)). We hypothesized that low activity COMT may be a risk factor for human breast cancer and designed a PCR-based RFLP assay to determine COMT genotype in a cohort of 112 matched, nested case-control samples. In the total study population, the odds ratios for the association of breast cancer risk with COMT(HL) and COMT(LL) genotypes were 1.30 [confidence interval (CI), 0.66-2.58] and 1.45 (CI, 0.69-3.07), respectively. Postmenopausal COMT(LL) women had a greater than 2-fold increased risk of developing breast cancer [odds ratio (OR), 2.18; CI, 0.93-5.11]. The association of COMT(LL) with the development of postmenopausal breast cancer was stronger and statistically significant in those women with a body mass index >24.47 kg/m2 (OR, 3.58; CI, 1.07-11.98). When COMT(LL) was combined with either glutathione S-transferase (GST) M1 null or with GSTP1 Ile-105-Val/Val-105-Val (intermediate/low activity, respectively) genotypes, the risk for developing postmenopausal breast cancer was also significantly increased. Our findings suggest that the allele encoding low activity COMT may be an important contributor to the postmenopausal development of breast cancer in certain women. PMID- 9407958 TI - eps15 and eps15R are essential components of the endocytic pathway. AB - eps15 and eps1SR are substrates of the epidermal growth factor (EGF) receptor kinase that are characterized by the presence of a protein:protein interaction domain, the EH domain, and by their ability to bind to the clathrin adaptor protein complex adaptor protein 2. Indirect evidence suggests that eps15 and eps15R are involved in endocytosis. Here we show that microinjection of antibodies against eps15 and eps15R inhibits internalization of EGF and transferrin. In addition, fragments of eps15 (encompassing its EH domains or the COOH-terminal region that binds to adaptor protein 2) inhibit EGF internalization or endocytosis of Sindbis virus. These results demonstrate that eps15 and eps15R are essential components of the endocytic machinery. PMID- 9407959 TI - Mitosin (a new proliferation marker) correlates with clinical outcome in node negative breast cancer. AB - Tumor proliferation rate is an important prognostic factor in breast cancer, and S-phase fraction (SPF), as measured by flow cytometry, is the most clinically validated of several methods for measuring it. However, flow cytometry is not well suited to evaluating the formalin-fixed, paraffin-embedded tumors that are routinely available or to the increasing number of small breast cancers. These and other limitations have motivated research into alternative methods for measuring proliferation, including immunohistochemistry (IHC) against cell cycle related antigens, which are better suited for the evaluation of small archival tissue samples. Mitosin is a recently described 350 kD nuclear phosphoprotein that is expressed in the late G1, S, G2, and M phases of the cell cycle but not in G0. Using a new monoclonal antibody (14C10), this pilot study evaluated mitosin expression by IHC in a series of 386 node-negative, formalin-fixed, archival breast cancers and correlated the results with several prognostic factors and clinical outcome (median follow-up, 78 months; range 3-214 months). The median and range of mitosin positive cells were 7% and 1-47%, respectively. There was a strong positive correlation between mitosin and SPF (r = 0.57; P = 0.0001), and there were significant negative correlations with estrogen receptor, progesterone receptor, and patient age. Mitosin was not related to overall survival in this pilot study. However, in a univariate cutpoint analysis of disease-free survival (DFS), patients with high levels of mitosin (>9% positive cells) had significantly worse DFS than did patients with lower levels (68% versus 84% at 5 years, respectively). In a multivariate analysis of DFS, large tumor size (>2 cm) and high mitosin were the only independently significant predictors of recurrence (relative risks = 2.47 and 1.72, respectively) in a model containing the additional factors estrogen receptor, progesterone receptor, patient age, and SPF. These preliminary results suggest that mitosin as assessed by IHC may be superior to SPF as a prognostic factor in node-negative breast cancer, but additional studies are necessary to validate these promising findings. PMID- 9407960 TI - Age-dependent tetrahydrothiophenium ion formation in young children and adults receiving high-dose busulfan. AB - Busulfan, a bifunctional alkylating agent, is a mainstay of myeloablative preparative regimens before hematopoietic stem cell transplantation. The apparent oral clearance of busulfan expressed relative to body surface area is 2-3-fold higher in children 1-4 years old than it is in adults. The first step in busulfan elimination is the formation of a tetrahydrothiophenium ion (THT+) in a glutathione S-transferase-catalyzed reaction. We present computer simulations that demonstrate that the ratio of the AUC of THT+ to that of busulfan over 6 h [(AUC(THT+)/AUC(BU))(0-->6)] is highly correlated (r2 = 0.805) with the determinants of THT+ formation and is virtually independent of the determinants of its elimination (r2 = 0.0201). We compared (AUC(THT+)/AUC(BU))(0-->6) determined in 14 children (0.5-4 years) to that of 11 adults (12-54 years) and found a 1.5-fold elevation in the area ratio (P = 0.0098) and a similarly significant increase in busulfan apparent oral clearance expressed relative to body surface area (P = 0.042). The only common explanation for the elevated busulfan apparent oral clearance and (AUC(THT+)/AUC(BU))(0-->6) is an enhanced ability of children to metabolize busulfan through glutathione conjugation. PMID- 9407961 TI - Aggressive breast cancer leads to discrepant serum levels of the type I procollagen propeptides PINP and PICP. AB - The propeptides PICP and PINP are derived from the synthesis of type I collagen, a major matrix protein of bone and soft tissues. The aim of this cross-sectional study was to investigate their value as indicators of the aggressivity of breast cancer. Serum PINP, PICP, and total alkaline phosphatase were determined from 89 breast cancer patients. Forty had major bone and/or soft tissue metastases with an aggressive disease course: the progressive disease (PD) group. Forty-nine had either none or minor bone and/or soft tissue metastases with a stable clinical course: the stable disease group (SD). The mean value of PINP in the PD group was 7.2 times higher than that in the SD group (276 +/- 79 microg/l versus 38 +/- 3 microg/l, respectively; P = 0.005), whereas PICP mean value was only 1.7 times higher in the PD group (174 +/- 20 microg/l versus 100 +/- 5 microg/l; P = 0.001). The ratio of PICP to PINP was 1.02 +/- 0.07 in the PD group and 3.07 +/- 0.18 in the SD group (P < 0.001). The correlation between PICP and PINP was linear in the SD group and nonlinear in the PD group. The results indicate that high serum PICP and PINP concentrations and a low PICP:PINP ratio are associated with a highly aggressive nature of breast cancer. Determination of PINP, in particular, may be valuable when evaluating the clinical status of a breast cancer patient. PMID- 9407962 TI - Effects of novel spermine analogues on cell cycle progression and apoptosis in MALME-3M human melanoma cells. AB - On the basis of encouraging preclinical findings, polyamine analogues have emerged as a novel class of experimental antitumor agents. The spermine derivative N1,N11-diethylnorspermine (DE-333, also known as DENSPM) is currently undergoing Phase I clinical trials against solid tumors. A series of systematically modified DE-333 analogues differing in intra-amine carbon distances and in N-alkyl terminal substituents (i.e., methyl, ethyl, and propyl) were evaluated in MALME-3M human melanoma cells, a cell line known to be cytotoxically affected by DE-333 and especially responsive to analogue induction of the polyamine catabolic enzyme spermidine/spermine N1-acetyltransferase. Analogues accumulated to comparable intracellular concentrations and similarly affected cell growth with IC50 values in the 0.5-1.0 microM range. During prolonged incubations, diethyl and dipropyl analogues were cytotoxic, whereas two dimethyl analogues were cytostatic. Cell cycle analysis following treatment with the cytotoxic analogues revealed a prominent G1 block apparent as an accumulation of cells in G0/G1 and depletion of S-phase cells as well as a less restrictive G2 block. By contrast, cytostatic analogues incompletely arrested cells in G1, leaving a significant number of S-phase cells. Morphological and immunocytochemical analysis of detached cells revealed a far greater proportion of apoptotic cells with cytotoxic analogues than with cytostatic analogues. Although spermidine/spermine N1-acetyltransferase activity was differentially induced by the analogues, there was no obvious correlation with cell cycle effects. Overall, these data indicate a previously unrecognized combined effect of polyamine analogues on cell cycle progression and apoptosis. On the basis of structure-function relationships, these activities may be manipulated to optimize therapeutic efficacy. PMID- 9407963 TI - Multicell spheroid response to drugs predicted with the comet assay. AB - Multicell spheroids were exposed to DNA-damaging agents with the aim of determining whether prompt DNA damage could be predictive for cell killing and drug resistance. Chinese hamster V79 cells, SiHa human cervical carcinoma cells, and WiDr human colon carcinoma cells were grown as spheroids and exposed to N methyl-N'-nitro-N-nitrosoguanidine (MNNG), 4-nitroquinoline-1-oxide (4NQO), doxorubicin, etoposide, actinomycin D, 1-(2-nitro-1-imidazolyl)-3-aziridino-2 propanol (RSU 1069), 3-amino-1,2,4-benzotriazine-1,4-dioxide (tirapazamine), and nitrogen mustard. Average DNA damage measured using the alkali comet assay generally correlated with cell killing irrespective of exposure times or drug concentration. However, better predictive power was achieved by using DNA damage levels in individual cells to identify the fraction of cells containing sufficient numbers of DNA strand breaks to cause death. Using this concept of a "threshold" for DNA damage, cell survival could be predicted for exposure to 4NQO, tirapazamine, nitrogen mustard, RSU 1069, and actinomycin D and was largely independent of cell type. The threshold value varied for each drug. For 4NQO, tirapazamine, and RSU 1069, DNA damage equivalent to about 10,000 strand breaks/cell was not toxic to cells of any spheroid type. Conversely, for actinomycin D, any DNA damage above background levels (approximately 100 breaks) was toxic for all three cell types. For some DNA-damaging drugs, the lack of correlation between DNA damage and cell killing was also informative. For etoposide and doxorubicin, no common threshold for cell killing could be determined, consistent with the hypothesis that DNA damage is only one of the actions of these drugs leading to cell death. For MNNG, the tail moment threshold varied significantly for the different spheroid types, probably indicating differences in repair. Overall, for five of the eight drugs, DNA damage measured using the comet assay was an effective and quantitative method of predicting drug cytotoxicity in complex multicelled systems. PMID- 9407964 TI - Autologous dendritic cells derived from CD34+ progenitors and from monocytes are not functionally equivalent antigen-presenting cells in the induction of melan A/Mart-1(27-35)-specific CTLs from peripheral blood lymphocytes of melanoma patients with low frequency of CTL precursors. AB - Peptide presentation by autologous dendritic cells (DCs) is a new tool to activate tumor antigen-specific T cells in melanoma patients. However, it is not known whether autologous DCs, differentiated by two of the most efficient protocols (from CD34+ progenitors or from monocytes), are equally effective as professional antigen-presenting cells (APCs) when the patients have a low frequency of peptide-specific precursors. To this end, a limiting dilution assay was applied to evaluate the frequency of antigen-specific CTL precursors (CTLps) in peripheral blood of HLA-A*0201+ melanoma patients. Then, from two melanoma patients showing low frequency of CTLps to melanoma antigen-A/melanoma antigen recognized by T cell (Melan-A/Mart-1)(27-35) peptide, autologous DCs were differentiated from granulocyte colony-stimulating factor-mobilized CD34+ progenitors or from monocytes. CD34+- and monocyte-derived DCs were characterized by a similar proportion of CD1a+ cells expressing HLA class II antigens and CD54, CD80, and CD86 molecules. Both types of DC presented Melan-A/Mart-1(27-35) and tyrosinase(369-377) peptides to melanoma-specific CTL clones and were equally effective as peptide-pulsed APCs in the activation of influenza A matrix(58-66) specific CTLs from high-frequency precursors (1294/10(6) and 1789/10(6) lymphocytes in the two patients). However, efficient activation of Melan-A/Mart 1(27-35)-specific CTLs from low-frequency precursors (158/10(6) and 77/10(6) lymphocytes) of the two patients was markedly dependent on the use of peptide loaded CD34+-derived DCs. These results suggest that CD34+- and monocyte-derived DCs are not functionally equivalent APCs for the activation of low-frequency peptide-specific CTLps. PMID- 9407965 TI - A transgenic mouse model with cyclin D1 overexpression results in cell cycle, epidermal growth factor receptor, and p53 abnormalities. AB - The cyclin D1 oncogene is critical in the progression of the cell cycle through the G1 phase. It is frequently overexpressed in squamous cell carcinomas originating from the head/neck and esophagus. Yet, the functional consequences of aberrant cyclin D1 overexpression are not entirely understood apart from increased cell proliferation. To address this question, we have developed a transgenic mouse model in which the EBV ED-L2 promoter targets cyclin D1 to the stratified squamous epithelium in a tissue-specific fashion to the tongue and esophagus, thereby resulting in a dysplastic phenotype. We now demonstrate that the dysplastic phenotype is associated with increased cell proliferation based on proliferating cell nuclear antigen overexpression and abnormalities in cyclin dependent kinase 4, epidermal growth factor receptor, and p53. In aggregate, these studies suggest that alterations in certain oncogenes and tumor suppressor genes occur early during head/neck and esophageal carcinogenesis. PMID- 9407966 TI - Adenovirus-mediated manganese superoxide dismutase gene transfer to hamster cheek pouch carcinoma cells. AB - As a first step in evaluating the tumor suppressor activity of the manganese superoxide dismutase (MnSOD) gene on established tumors in vivo, we used adenovirus-mediated gene transfer as a means of delivering the MnSOD cDNA to hamster cheek pouch carcinoma (HCPC-1) cells in vitro. HCPC-1 cells were transduced with the adenovirus-MnSOD construct (AdMnSOD) at multiplicities of infectivity (MOI) of 0, 10, 25, 50, 100, 150, and 200 MOI or with the adenovirus LacZ reporter gene construct (AdLacZ) at 100 MOI. Dose-dependent increases in MnSOD immunoreactivity were seen on Western blotting and indirect immunofluorescence microscopy with increasing AdMnSOD titers. Maximal immunoreactivity was observed at 100 MOI AdMnSOD with both techniques. Moreover, we observed a concomitant 6-7-fold increase in MnSOD activity compared with parental cell levels that also peaked at 100 MOI AdMnSOD. To determine the effect of transgene-expressed MnSOD on tumor cell behavior, we examined cell growth, plating efficiency, and anchorage-dependent growth in soft agar. Cell number measured on day 13 decreased approximately 50% with 100 MOI AdMnSOD (P < 0.05) compared with parental cells. Moreover, cell doubling time increased from 38 to 44 h with 100 MOI AdMnSOD. Plating efficiency and cell growth in soft agar decreased approximately two-thirds with 100 MOI AdMnSOD (P < 0.001). These assays of the transformed phenotype in vitro all appeared to show maximal effect with 100 MOI AdMnSOD. As tumor growth in vivo is most predictable by a combination of these in vitro data, our results suggest that if MnSOD can be effectively delivered to a tumor in vivo using the adenovirus paradigm, effective tumor growth suppression can be observed. PMID- 9407967 TI - Chromosomal instability and its relationship to other end points of genomic instability. AB - Chromosomal destabilization is one end point of the more general phenomenon of genomic instability. We previously established that chromosomal instability can manifest in clones derived from single progenitor cells several generations after X-irradiation. To understand the potential relationship between chromosomal destabilization and the other end points of genomic instability, we generated a series of chromosomally stable and unstable clones by exposure to X-rays. All clones were derived from the human-hamster hybrid line GM10115, which contains a single copy of human chromosome 4 in a background of 20-24 hamster chromosomes. These clones were then subjected to a series of assays to determine whether chromosomal instability is associated with a general "mutator phenotype" and whether it modulates other end points of genomic instability. Thus, we analyzed clones for sister chromatid exchange, delayed reproductive cell death, delayed mutation, mismatch repair, and delayed gene amplification. Statistical analyses performed on each group of chromosomally stable and unstable clones indicated that, although individual clones within each group were significantly different from unirradiated clones for many of the end points, there was no significant correlation between chromosomal instability and sister chromatid exchange, delayed mutation, and mismatch repair. Delayed gene amplification was found to be marginally correlated to chromosomal instability (P < 0.1), and delayed reproductive cell death (the persistent reduction in plating efficiency after irradiation) was found to be significantly correlated (P < 0.05). These correlations may be explained by chromosomal destabilization, which can mediate gene amplification and can result in cellular lethality. These data implicate multiple molecular and genetic pathways leading to different manifestations of genomic instability in GM10115 cells surviving exposure to DNA-damaging agents. PMID- 9407968 TI - Transgenic mice overexpressing a dominant-negative mutant type II transforming growth factor beta receptor show enhanced tumorigenesis in the mammary gland and lung in response to the carcinogen 7,12-dimethylbenz-[a]-anthracene. AB - To test the hypothesis that the transforming growth factor-beta (TGF-beta) system has tumor suppressor activity in the mammary gland, we have generated transgenic mice overexpressing a dominant-negative mutant form of the type II TGF-beta receptor, under the control of the mouse mammary tumor virus-long terminal repeat. High-level expression of the transgene was observed in the mammary and salivary glands, with lower expression in the lung, spleen, and testis. Older nulliparous transgenic mice (9-17 months) showed a marked increase in the incidence and degree of lobulo-alveolar side-branching in the mammary glands when compared to wild-type littermates (24.8% of glands examined histologically versus 14.4%; P = 0.004), suggesting a role for endogenous TGF-betas in regulating development or maintenance of mammary alveoli. Spontaneous tumorigenesis was unchanged in the transgenic mice. However, following initiation with the carcinogen 7,12-dimethylbenz[a]anthracene, the transgenic group showed a significant increase in the incidence and multiplicity of mammary tumors when compared with wild-type littermates (40% incidence in transgenic mice versus 22% for wild-type, with 4 of 25 transgenics developing multiple mammary tumors versus 0 of 27 wild-type; P = 0.03). An early increase in the incidence of lung tumors was also observed in transgenic mice, but no difference between genotype groups was seen in the incidence of tumors in tissues in which the transgene is not expressed. The data show that the endogenous TGF-beta system has tumor suppressor activity in the mammary gland and lung. PMID- 9407969 TI - Fas/APO-1 (CD95) is not translocated to the cell membrane in esophageal adenocarcinoma. AB - This study describes Fas (CD95) expression in Barrett's esophagus, adenocarcinomas of the esophagus, and three esophageal adenocarcinoma cell lines. Immunohistochemical analysis of Barrett's esophagus demonstrated cell surface expression of Fas protein. In contrast, 30.5% of esophageal adenocarcinomas examined by immunohistochemical analysis demonstrated faint cytoplasmic staining, and 69.5% were negative for Fas. Similar levels of Fas mRNA were identified in tumors compared to mRNA levels in esophageal squamous mucosa or Barrett's esophagus. An approximately Mr 48,000 Fas protein was identified by Western blot analysis in tumors that were negative for Fas expression by immunohistochemical analysis. The esophageal adenocarcinoma cell line Seg-1 was negative for Fas expression by immunohistochemical analysis, but Western blot analysis demonstrated abundant, appropriately sized Fas protein. In agreement with the immunohistochemical analysis, flow cytometry of Seg-1 showed minimal amounts of Fas on the cell surface, which correlated with resistance to Fas-mediated apoptosis. No mutations in the Seg-1 Fas coding sequence or exon 1 were identified by sequence analysis. This was confirmed by transient transfection of COS cells with expression vectors generated from the Seg-1 Fas cDNA, which resulted in cell surface expression of the Fas protein. Stable transfection of Seg-1 with a Fas expression vector did not result in efficient Fas expression on the cell surface. Seg-1 cells, transiently transfected with a Fas-FLAG expression vector and examined for protein expression using confocal microscopy and an anti FLAG antibody, showed that the Fas-FLAG protein was not present on the cell surface but was present in the cytoplasm. Taken together, these results indicate that expression of Fas on the cell surface by esophageal adenocarcinoma is reduced. In an esophageal adenocarcinoma cell line, wild-type Fas protein is retained in the cytoplasm, and this correlates with resistance to Fas-mediated apoptosis. The retention of wild-type Fas protein within the cytoplasm may represent a mechanism by which malignant cells evade Fas-mediated apoptosis. PMID- 9407970 TI - Expression of sex hormone-binding globulin exon VII splicing variant messenger RNA in human uterine endometrial cancers. AB - We have demonstrated the intracellular expression of sex hormone-binding globulin (SHBG) exon VII splicing variant mRNA in human uterine endometrial cancer using the reverse transcription-PCR-Southern blot and DNA sequencing analyses. Analysis of the missing base pairs proved that they corresponded to the entire exon VII, which is considered to encode a portion of the steroid-binding site, suggesting that the steroid-binding affinity of this variant might be different from that of the SHBG wild type. In uterine endometrial cancers, the wild-type mRNA levels significantly (P < 0.01) decreased, and the ratio of the SHBG variant to wild type mRNA levels (P < 0.01) increased with the advance of histological dedifferentiation. These results suggest that dedifferentiation of endometrial cancers might induce a reduction in their estrogen-dependent properties via intracellular SHBG. PMID- 9407971 TI - Abrogation of wild-type p53-mediated transactivation is insufficient for mutant p53-induced immortalization of normal human mammary epithelial cells. AB - The p53 protein has become a subject of intense interest since the discovery that about 50% of human cancers carry p53 mutations. Mutations in the p53 gene are the most frequent genetic lesions in breast cancer, suggesting a critical role for p53 protein in normal mammary epithelial cell (MEC) growth control. We previously demonstrated that abrogation of the p53 function by a cancer-derived p53 mutant, del239, was sufficient to induce immortalization of normal MECs. To further extend these findings and to examine the mechanism of mutant p53-induced immortalization of MECs, we tested the immortalizing ability of four selected p53 mutants (R175H, R248W, R249S, and R273H), which involve residues that cluster close to N239 in the three-dimensional structure and which are critical for the DNA-binding function of p53. Interestingly, two of these mutants (R175H and R249S) reproducibly immortalized 76N normal MECs, whereas the other two mutants (R248W and R273H) induced an extension of life span but not immortalization. These results further substantiate that selective ablation of p53 function with dominant-negative mutants is sufficient for immortalization of MECs. To determine whether abrogation of the transactivation function of endogenous p53 was important for the differential immortalizing ability of p53 mutants, we measured the effects of mutant p53 on the endogenous wild-type p53-mediated transactivation of a chloramphenicol acetyltransferase reporter linked to a consensus p53 binding DNA sequence in transiently transfected 76N MECs. All of the mutants, regardless of their immortalizing phenotype, abrogated the endogenous wild-type p53-mediated transactivation to a similar extent. Thus, abrogation of transactivation function is not sufficient for mutant p53-induced immortalization of normal MECs. The p53-immortalized MECs showed substantial telomerase activity; however, induction of telomerase activity occurred at late passages and was undetectable in mutant p53-expressing cells prior to immortalization. We suggest that mechanisms other than abrogation of transactivation and induction of telomerase activity determine the differential MEC-immortalizing behavior of various p53 mutants. PMID- 9407972 TI - Angiogenesis by fibroblast growth factor 4 is mediated through an autocrine up regulation of vascular endothelial growth factor expression. AB - The infection of normal mouse mammary EF43 cells by a retroviral vector carrying either Fgf-3 (EF43.Fgf-3) or Fgf-4 (EF43.Fgf-4) cDNA resulted in the transformation of cells displaying different tumorigenic potentials in nude mice (A. Hajitou and C-M. Calberg-Bacq, Int. J. Cancer, 63: 702-709, 1995). EF43.Fgf-4 produced rapidly developing tumors at all sites of inoculation, whereas EF43.Fgf 3 produced slowly growing tumors only in the mammary fat pad. Cells infected with the vector carrying the selection gene alone (EF43.C) were not tumorigenic. The angiogenic properties of these cells were tested in an in vitro angiogenesis model using human umbilical vein endothelial cells (HUVECs) cultured at the surface of a type I collagen gel and their capacity to form tube-like structures on invasion of the gel. Only the conditioned medium (CM) of EF43.Fgf-4 induced an angiogenic morphotype in HUVECs. In parallel, the mRNA expression of matrix metalloproteinase 1 and c-ETS-1 was increased in the HUVECs displaying a differentiated phenotype, whereas the tissue inhibitor of matrix metalloproteinase 1 mRNA level was decreased. Recombinant human fibroblast growth factor 4 (FGF-4) did not induce an angiogenic phenotype in HUVECs by itself. By Western blot analysis, a high expression of vascular endothelial growth factor (VEGF) was detected in the EF43.Fgf-4 CM. This result was confirmed by Northern blot analysis of total RNA extracted from the three cell types; the steady-state level of VEGF mRNA was low and equivalent in EF43.C and EF43.Fgf-3, whereas it was strongly increased in EF43.Fgf-4. Culturing EF43 cells carrying only the selection gene with increasing concentrations of recombinant human FGF-4 resulted in a dose-dependent stimulation of VEGF. The induction of the angiogenic morphotype and the parallel modulations of the biosynthetic phenotype in HUVECs were completely suppressed by adding a neutralizing antibody directed against VEGF to EF43.Fgf-4 CM. Furthermore, inhibition of protein kinase C by bisindoylmaleimide suppressed the angiogenic phenotype induced by the CM of EF43.Fgf-4. Our results point to an indirect angiogenic activity of FGF-4 through the autocrine induction of VEGF secretion by EF43.Fgf-4 cells, an original signaling pathway that might be significant in tumor progression and metastasis. PMID- 9407973 TI - Cell scattering and migration induced by autocrine transforming growth factor alpha in human glioma cells in vitro. AB - In the present investigation, we have transfected a human malignant glioma cell line, U-1242 MG, and derived clones that produce transforming growth factor alpha (TGF-alpha) in an inducible manner using the tetracycline suppressible vector system. TGF-alpha expression was confirmed by Northern analysis, by ELISA, and by immunoprecipitation of metabolically labeled cells. The functional activity of the induced protein was proven by the finding of epidermal growth factor receptor (EGFR) tyrosine phosphorylation on induction of TGF-alpha. A clear effect on cell motility, i.e., cell scattering and an increased phagokinetic track area of individual glioma cells, was demonstrated. The fact that the EGFR tyrosine kinase activation was independent of cell density suggests that autocrine activation of the EGFR kinase occurred at the single-cell level. These findings are of interest, because increased cell motility is most likely a requirement for glioma cell invasion in vivo. The results imply that as a result of coexpression of EGFR and its ligand, individual glioma cells are capable of acting as independent autocrine locomotory units. PMID- 9407975 TI - Correspondence re: G. Palumbo et al., the tyrphostin AG17 induces apoptosis and inhibition of cdk2 activity in a lymphoma cell line that overexpresses bcl-2. Cancer Res., 57: 2434-2439, 1997. PMID- 9407974 TI - Telomerase activity in the normal and neoplastic rat mammary gland. AB - The 1-methyl-1-nitrosourea-induced rat mammary tumor model system is well studied, reproducible, and widely used. We have investigated whether these tumors possess higher telomerase activity than normal mammary tissue. Using the telomeric repeat amplification protocol assay, we found significantly higher telomerase activity in 36 mammary carcinomas than in 72 mammary glands of virgin rats. The level of telomerase activity in virgin rats was unaffected by strain, age, stage of the estrous cycle, or ovariectomy. However, mammary glands obtained from pregnant rats exhibited telomerase activity comparable to that found in the tumors, possibly reflecting the high epithelial content of these tissues. Indeed, isolated epithelial cells from virgin and pregnant mammary glands and from carcinomas had similar telomerase activities. Thus, telomerase activity is constitutive in the rat mammary epithelium and is not a unique characteristic of malignant transformation in this tissue. These results underscore the importance of attributing biochemical properties to specific cell types in a tissue, a situation not paralleled in the interpretation of data from in vitro models. PMID- 9407976 TI - Prospective, randomized study comparing clinical results between small and large colonic J-pouch following coloanal anastomosis. AB - PURPOSE: Improved functional results can be obtained by construction of a colonic J-pouch after coloanal anastomosis. Variability in pouch size following coloanal anastomosis is prevalent in current literature. In this study, the authors compare clinical bowel function after complete rectal excision with coloanal anastomosis for patients with rectal carcinoma using either a small 6-cm or a large 10-cm colonic J-pouch anastomosis. The clinical outcome is assessed both at short-term and long-term follow-up. METHODS: Fifty-nine consecutive patients with rectal cancers 4 to 8 cm from the anal verge were recruited into the study. Patients were randomized intraoperatively to either a 6-cm J-pouch group or a 10 cm J-pouch group. Clinical assessments were performed prospectively at 3, 6, 12, and 24 months postoperatively, following colostomy closure. Clinical parameters such as frequency, urgency, continence, and laxative and enema use were assessed and compared between the two groups. RESULTS: There was no statistical differences in the mean defecation frequency, urgency, and fecal continence between the two groups at 3, 6, 12, and 24 months. In the first year, laxative and enema use between the two groups was negligible; however at two years, 30 percent of patients with a large reservoir compared with 10 percent of patients in the small-pouch group required laxative and/or enema for constipation and evacuation of bowels. CONCLUSION: Similar clinical results can be expected from patients with either small or large reservoirs at one year. However, with long term follow-up, patients with a large reservoir are more likely to require medication for constipation and evacuation. To avoid these inconveniences a small reservoir is advocated for patients undergoing coloanal anastomosis. PMID- 9407978 TI - Surgical treatment of locally recurrent rectal carcinoma. AB - PURPOSE: This study was performed to analyze the outcomes of patients with local (pelvic) recurrence (following radical surgery for rectal cancer) who subsequently underwent a new operation. METHODS: Forty-five patients (19 percent of 213 local recurrences) were explored surgically because the disease was deemed to be confined to the pelvis with a limited extension and, therefore, amenable to surgical cure. RESULTS: Only 21 of the 45 patients who underwent surgical exploration had an oncologically radical operation (R0). In the remaining 24 patients, either a simple exploration or palliation or a nonradical procedure (R1 R2) was performed. In the R0 group, there was a 19 percent five-year survival rate vs. a 0 percent rate in the R1-R2 group (median survival, 4 months). Site of recurrence (anastomosis vs. other sites) was statistically associated with a higher chance of long-term survival for those who underwent an R0 operation. CONCLUSIONS: The prognosis of locally recurrent rectal cancer is dismal; less than 10 percent of all patients who underwent surgical treatment benefit from reoperation with an overall survival for five years. On the basis of these results, we no longer consider the surgical approach as the primary option for treating locally recurrent rectal cancer. PMID- 9407977 TI - Suppository administration of chemotherapeutic drugs with concomitant radiation for rectal cancer. AB - PURPOSE: Preoperative radiation with combined chemotherapy is effective in shrinking advanced rectal cancer locally and facilitating subsequent surgery. Suppository delivery of 5-fluorouracil is associated with less toxicity and higher rectal tissue concentrations than intravenous administration. This prompted us to evaluate suppository and intravenous administration of 5 fluorouracil and mitomycin C with concomitant radiation to determine associated toxicity. METHODS: Rectal, liver, lymph node, and lung tissue and systemic and portal blood were collected serially from male Sprague Dawley rats to determine drug concentrations following suppository or intravenous delivery of 5 fluorouracil or mitomycin C. Thirty-six animals were randomly assigned to treatment groups and received 5-fluorouracil suppositories, mitomycin C suppositories, or an equivalent intravenous dose of 5-fluorouracil or mitomycin C 30 minutes before radiation therapy. Before and 3, 6, 10, and 15 days following this treatment, blood was collected, colonoscopy was performed, and rectal tissue was harvested for histologic examination. RESULTS: Mitomycin C suppository was significantly less toxic compared with intravenous delivery, and higher rectal tissue concentrations were observed from 10 to 30 minutes (P < 0.05). Compared with intravenous 5-fluorouracil administration and radiation, 5-fluorouracil suppository and radiation resulted in additive myelosuppression at day 6 (P < 0.05) with rapid recovery. CONCLUSIONS: 5-Fluorouracil and mitomycin C suppository delivery combined with radiation causes less systemic toxicity and is more effective than intravenous administration. PMID- 9407979 TI - Multivariate analysis of the prognostic factors of patients with unresectable synchronous liver metastases from colorectal cancer. AB - PURPOSE: It frequently is observed that widely varying prognoses are given for patients with the same extent of liver metastases from colorectal cancer, even though the same treatment is performed on these patients. One of the reasons for this variance is that prognostic factors for these patients have not been defined. This study was designed to elucidate which clinicopathologic factors were the most important in the prognosis of 73 patients with unresectable synchronous liver metastasis from colorectal cancer. METHODS: Univariate and multivariate analysis of 11 clinicopathologic factors were performed using the Cox proportional hazard model. Survival curves were generated using the Kaplan Meier method. RESULTS: Extent of liver metastases was the most significant variable in this survival analysis, although the extent of lymph node metastases of the primary lesion also was significant. However, the method of treatment was not a significant determinant in the survival for patients with unresectable liver metastases. Median survival of patients with H1, H2, and H3 was 13, 12, and 6 months, respectively, and there was a significant difference between survival curves for patients with H1 and patients with H3. Median survival of patients with n0, n1, and n2 was 13, 7, and 7 months respectively, and there was a significant difference between survival curves for patients with n0 and patients with n2. Median survival of 6 patients with H1 and n0 and of 17 patients with H3 and n2 was 28 and 4 months, respectively. There was a significant difference in survival curves between these two groups. CONCLUSION: Longevity of patients with unresectable synchronous liver metastases from colorectal cancer is affected adversely by the presence of nodal metastases and extent of liver metastases. This should be considered in the planning treatment. PMID- 9407980 TI - Diagnosing anal sphincter injury with transanal ultrasound and manometry. AB - PURPOSE: This study was undertaken to evaluate how well anorectal manometry and transanal ultrasonography diagnose anal sphincter injury. METHODS: Anorectal manometry and transanal ultrasonography were performed in 20 asymptomatic nulliparous women and 20 asymptomatic parous women, and the results were compared with those obtained in 31 incontinent women who subsequently underwent sphincteroplasty and, thus, had operatively verified anal sphincter injury. By using computerized manometry analysis, mean maximum resting and squeeze pressures, sphincter length, and vector symmetry were determined in all women. All transanal ultrasounds were interpreted blinded as to the patient's history, physical examination, and manometry results. RESULTS: Manometric resting and squeeze pressures were significantly higher in the asymptomatic nulliparous women than in the asymptomatic parous women, and both groups had significantly higher pressures than the incontinent women (P < 0.001). Anal sphincter length and vector symmetry index were significantly decreased in incontinent women compared with asymptomatic women (P < 0.01). Decreased resting and squeeze pressures suggestive of possible sphincter injury were found in 90 percent of incontinent women with known anal sphincter injury. Decreased anal sphincter length and vector symmetry were found in only 42 percent of women with known anal sphincter injury. Transanal ultrasound was able to identify 100 percent of the known sphincter injuries but also falsely diagnosed injury in 10 percent of the asymptomatic nulliparous women with intact anal sphincters. False identification of sphincter injury increased when transanal ultrasound scanning was performed proximal to the distal 1.5 cm of the anal canal. CONCLUSION: Although nonspecific, decreased resting and squeeze pressures were found in 90 percent of patients with anal sphincter injury. Decreased anal sphincter length or vector symmetry index were present in only 42 percent of patients with known sphincter injury. When limited to the distal 1.5 cm of the anal canal, transanal ultrasound identified all known sphincter injuries but falsely identified injury in 10 percent of women with intact anal sphincters. Transanal ultrasound in combination with decreased anal pressures correctly identified all intact sphincters and 90 percent of known anal sphincter injuries. PMID- 9407981 TI - Randomized controlled trial of primary fistulotomy with drainage alone for perianal abscesses. AB - PURPOSE: Primary fistulotomy may be advantageous for perianal abscesses because unlike ischiorectal abscesses, fistulas are more commonly found and can be laid open with full preservation of the external anal sphincters. Therefore, a randomized, controlled trial was conducted to compare primary fistulotomy with incision and drainage alone, specifically for perianal abscesses. METHODS: Fifty two consecutive patients (43 males; mean age, 40 (standard error of mean, 2) years) with perianal abscesses were randomized to treatment by either incision and drainage (controls; N = 28) or fistulotomy (N = 24). Patients were followed up clinically for a mean of 15.5 (standard error of the mean, 0.7) months. Anorectal manometry was also performed before, six weeks, and three months after surgery. RESULTS: Persistent fistulas developing after surgery were significantly more common after incision and drainage (N = 7; 25 percent) than after fistulotomy (N = 0; P = 0.009). One patient in each group was also found to have a residual abscess, which required repeat drainage. All patients remained fully continent. The anal pressures after incision and drainage and fistulotomy were not significantly different. Operative time, hospital stay, and time for the wound to heal completely were the same in both groups. CONCLUSIONS: Primary fistulotomy at the time of drainage for perianal abscesses results in fewer persistent fistulas and no added risk of fecal incontinence. PMID- 9407982 TI - Tailored lateral sphincterotomy for anal fissure. AB - PURPOSE: Most surgical texts describe the length of division of the internal sphincter during closed lateral sphincterotomy as "to just above the dentate line," resulting in significant rates of incontinence. This study reviews our experience using a "tailored" lateral sphincterotomy by selecting the height of sphincter to be divided with the aim of preserving more sphincter. METHODS: From 1976 to March 1996, the files of 440 patients who had sphincterotomies were reviewed by an independent research assistant. After exclusions, a residual group of 352 patients had undergone tailored left lateral sphincterotomy for chronic anal fissure that had failed conservative treatment or for acute anal fissure requiring surgical intervention. RESULTS: A total of 287 patients from the group who had tailored left lateral sphincterotomy returned for review (81.5 percent). Of these, four complained of imperfect control of flatus (1.4 percent), one of minor staining (0.35 percent), and two of urgency (0.7 percent). None had incontinence of feces or leakage of stool. Five patients had repeat sphincterotomies, four for recurrence and one for a persistent fissure. CONCLUSION: The technique of tailored lateral sphincterotomy is safe, effective, and preserves more anal sphincter. It might be argued that a controlled trial comparing tailored sphincterotomy with the standard height of incision (with preprocedure and postprocedure manometry) should be performed, but the clinically significant reduction in incontinence rates using the tailored approach would seem to support its use. PMID- 9407983 TI - Cutting seton for anal fistulas: high risk of minor control defects. AB - PURPOSE: Long-term results of cutting seton in the treatment of anal fistulas were studied. METHODS: Of the 44 patients with anal fistulas, mainly of the high variety, managed with this method, 35 (25 men) attended a clinical and manometric follow-up examination on average 70 (range, 28-184) months after operation. Fistula distribution was high transsphincteric (25), low transsphincteric (5), extrasphincteric (3), and suprasphincteric (2). The seton was tightened at one week to two-week intervals to achieve gradual sphincter division. RESULTS: Time required to achieve complete fistula healing ranged from 37 to 557 (mean, 151) days. Two (6 percent) of the 35 patients re-examined had recurrence of fistula and 22 (63 percent) reported symptoms of minor impairment in anal control, which in four patients had existed already before operation. Anal resting pressures were similar for defective and normal control, but other manometric variables were inferior in incontinence, although total squeeze pressure only showed statistically significant difference from normal continence (P = 0.0345). Incontinence was likely associated with hard and gutter-shaped operation scars in the anal canal, but the difference from normal continence was not statistically significant. CONCLUSION: Cutting seton yields fairly good results in regard to cure of fistula, but the risk of anal incontinence, despite its minor degree, seems to be too high to recommend its routine use for all high fistulas. The suprasphincteric fistulas and some extrasphincteric fistulas are difficult to treat otherwise, but especially for high transsphincteric fistulas, other methods of treatment (preferably those in which sphincter division can be avoided and the risk of anal canal deformity and incontinence are minimized) are advocated. PMID- 9407984 TI - Outcome of patients with total colonic ischemia. AB - PURPOSE: In this study, we sought to determine the outcome of patients with ischemic colitis, comparing patients with segmental disease with those with total colonic ischemia. METHODS: Patients with the diagnosis of ischemic colitis over the past six years were selected and reviewed for demographics, presenting symptoms, diagnosis, and treatment. RESULTS: Forty-three consecutive patients with ischemic colitis were identified and were grouped into those with segmental ischemic colitis and total colonic ischemia. Mean age was 68.8 years; 28 of 43 patients (65 percent) were males. Diagnosis was established by colonoscopy in 31 of 43 patients (72 percent), whereas in the remainder, diagnosis was made in the operating room. Ischemic colitis developed in the hospital in 17 of 43 patients (40 percent) during admission for an unrelated illness. In 6 of 43 (14 percent) of these patients, ischemic colitis developed following surgery. Thirty-one of 43 patients (72 percent) were found to have segmental colitis; 11 of 31 patients (35 percent) were successfully managed nonoperatively. Segmental colitis was present in 31 of 43 patients (72 percent), and 12 of 31 (35 percent) of these patients were successfully managed nonoperatively. In the patients with segmental colitis who required surgery, the 30-day mortality rate was 22 percent. Among 12 of 17 patients (71 percent) with segmental ischemia treated by resection and stoma, 9 of 12 (75 percent) underwent eventual stoma closure. All 12 patients with total colonic ischemia required surgery, and 9 of 12 patients (75 percent) died. CONCLUSION: Ischemic colitis occurs commonly during an unrelated hospital admission and following previous surgery. Most patients treated by resection and stoma undergo stoma closure. Total colonic ischemia carries a worse prognosis than segmental colonic ischemia. PMID- 9407985 TI - Total abdominal colectomy and ileorectal anastomosis for inflammatory bowel disease. AB - PURPOSE: This retrospective study assesses the results of total colectomy and ileorectostomy for inflammatory bowel disease. METHODS: Between January 1974 and December 1990, 90 patients underwent total colectomy and ileorectal anastomosis for chronic ulcerative colitis (n = 48) or Crohn's colitis (n = 42) at the Mayo Clinic. Patients' records were reviewed retrospectively. Long-term results were assessed by chart reviews and postal questionnaires. Conversion to a permanent ileostomy, with or without proctectomy, was considered a failure of the procedure. The Kaplan-Meier method was used to estimate survivorship free of failure. The log-rank test was used to compare survivorship curves. Ninety-five percent confidence intervals were calculated at selected time points. P values < 0.05 were considered to be statistically significant. RESULTS: The main indication for surgery was refractory chronic disease. There were no immediate postoperative deaths. The anastomotic leakage rate was 4.4 percent, and small bowel obstruction occurred in 15.6 percent. At the time of follow-up (mean, 6.5 +/- 4.8 years), 46 patients (58.9 percent) had recurrence or exacerbation of the disease. This was the most common indication for subsequent proctectomy/permanent ileostomy in the follow-up period. There were 8 failures in 48 patients with ulcerative colitis (16.7 percent) and 11 failures in 42 patients with Crohn's disease (26.2 percent), although this difference was not statistically significant. Cumulative probability of having a functioning ileorectal anastomosis at five years was 84.2 percent (95 percent confidence interval, 71 95.9 percent) for ulcerative colitis and 73.8 percent (95 percent confidence interval, 58.6-88.6 percent) for Crohn's disease. In the latter group, females showed a significantly lower cumulative probability of having a functioning ileorectal anastomosis (females, 63.4 percent; males, 92.3 percent; P = 0.04). Crohn's patients 36 years of age or younger also showed a lower probability of success (patients < or = 36 years, 57 percent; patients > 36 years, 93.8 percent; P = 0.03). In the group with chronic ulcerative colitis, younger patients also seemed to require additional surgery more frequently; however, this difference was not statistically significant. Previous duration of symptoms, with mild or moderate disease in a distensible rectum, had no effect on results in either disease group. Functional results were acceptable in 63.6 and 87.5 percent of patients with Crohn's and ulcerative colitis, respectively. Eighty-four percent of ulcerative colitis patients and 91 percent of Crohn's disease patients reported an improvement in their quality of life, and overall, more than 90 percent considered their health status to be better than before surgery. One patient with ulcerative colitis developed carcinoma of the rectal stump 11.5 years after the colectomy and ileorectal anastomosis (cumulative probability of remaining free of cancer, 85.7 percent at 12 years; 95 percent confidence interval, 57.7-100 percent). CONCLUSIONS: These results demonstrate that, in selected patients with a relatively spared rectum and without severe perineal disease, total colectomy and ileorectal anastomosis still remains a viable option to total proctocolectomy with extensive Crohn's colitis. In addition, ileorectal anastomosis, as a sphincter-saving procedure, continues to have a place in the surgical treatment of chronic ulcerative colitis for high-risk or older patients who are not good candidates for ileal pouch-anal anastomosis, when the latter procedure cannot be done because of technical reasons and in the presence of advanced carcinoma concomitant with colitis, when life expectancy is limited. PMID- 9407986 TI - Prognostic relevance of occult tumor cells in lymph nodes of colorectal carcinomas: an immunohistochemical study. AB - PURPOSE: Whereas lymph node metastases in colorectal carcinoma are an important prognostic factor, the prognostic relevance of occult tumor cells in lymph nodes is not elucidated at present. Therefore, our study intended to assess the rate of patients with occult tumor cells in histopathologically negative lymph nodes. Furthermore, we tried to evaluate an eventual influence of these occult tumor cells on patients' prognoses. METHODS: For examination, we used paraffin blocks of lymph nodes, tumor-negative by conventional histopathology, from 49 patients with colorectal carcinoma (Stage I-III) after a curative (R0) tumor resection in 1987. After preparation of tissue blocks using the serial sectioning technique, the specimens were stained with the alkaline phosphatase, antialkaline phosphatase method and two monoclonal antibodies (AE1/AE3 and Ber-EP4). RESULTS: In 13 of 49 patients (26.5 percent), we disclosed tumor cells, mostly located in subcapsular sinuses as single cells or in groups. There was a good correlation between the detection rate and N category, tumor stage, and grading. Moreover, 33 percent of patients in Stage I/II with occult tumor cells (N0+) developed a local relapse and/or distant metastases in contrast to 12 percent of patients without tumor cells (N0-). With a median follow-up of 84 months, we found no difference in disease-free survival between the tumor cell negative and positive groups in Stage I/II patients. CONCLUSION: The results show that occult tumor cells might increase the risk for development of a local tumor relapse and/or distant metastases but do not influence patients' prognoses at all. PMID- 9407987 TI - Lymph node involvement and tumor depth in rectal cancers: an analysis of 805 patients. AB - BACKGROUND: Superficial rectal tumors are said to involve regional lymph nodes rarely. This presumption must be proven beyond any doubt if less radical surgery is to be offered for such patients. PATIENTS AND METHODS: Eight hundred five cases (467 males; median age, 64 (range, 19-97) years) of rectal cancer were reviewed. RESULTS: Lymph node positivity, number of lymph nodes involved, lymphatic vessel, and venous and perineural invasion were significantly increased with increasing depth of invasion of tumor through the bowel wall in univariate analysis. The percentage of lymph node involvement at each tumor depth was as follows: T1, 5.7 percent; T2, 19.6 percent; T3, 65.7 percent; T4, 78.8 percent. Overall lymph node involvement was 59 percent. For patients younger than 45 years of age, the percentage of lymph node involvement was 33.3, 30, 69.3, and 83.3 percent compared with 3.1, 8.4, 64.2, and 78.8 percent for patients aged 45 years or above for T1, T2, T3, and T4, respectively. CONCLUSION: Increased depths of tumor penetration beyond T1 and age less than 45 years have an excessive incidence of lymph node positivity. The finding of lymphatic vessel invasion on biopsy is highly indicative of lymph node metastasis. PMID- 9407988 TI - Pelvic anatomy and pathology is influenced by distention of the rectum: defecoperitoneography before and after rectal filling with contrast medium. AB - PURPOSE: The aim of the present study was to evaluate how distention of the rectum with contrast medium at defecoperitoneography affected pelvic anatomy, i.e., position, form, and size, of organs and pouch of Douglas. PATIENTS AND METHODS: Twenty-six female patients with a peritoneocele at defecoperitoneography were selected for the present study. Radiographs taken at the start, before, and after filling the rectum with contrast medium were compared. RESULTS: There was an obvious change in the position of the organs in the pelvis when the rectum was distended with contrast medium. The peritoneocele disappeared completely in 19 of the patients and was reduced in size in the remaining 7 patients, and the enterocele disappeared completely in 13 patients. The small bowel and vaginal portion of the uterus moved cranially. CONCLUSIONS: A distended rectum may conceal existing pathology, such as peritoneocele and enterocele, at defecoperitoneography. Defecoperitoneography should, therefore, include a radiograph before the rectum is filled. This radiograph shows the habitual anatomy of the patient in the sitting position and may demonstrate pathologic findings. PMID- 9407989 TI - Aspirin effects on colonic mucosal bleeding: implications for colonic biopsy and polypectomy. AB - BACKGROUND: Many patients who require endoscopic treatments such as biopsy and polypectomy are given antiplatelet agents reluctantly. We have studied the effects of aspirin on colonic mucosal hemostasis. METHODS AND PATIENTS: We developed a new endoscopic device to make a standard incision (7-mm length) on the colonic mucosa to study colon bleeding time. We measured the colon bleeding time of normal colonic mucosa in 47 cases. The colon bleeding time and skin bleeding time (Simplate method) were measured before and one hour after aspirin ingestion (990 mg) in ten healthy subjects. RESULTS: The bleeding time of normal colonic mucosa was 156 +/- 71 (mean +/- standard deviation) seconds. Significant prolongation was noted in both skin bleeding time (357 +/- 192 vs. 477 +/- 183 seconds; P < 0.05) and colon bleeding time (155 +/- 47 vs. 244 +/- 169 seconds; P < 0.05) after aspirin ingestion. CONCLUSIONS: Bleeding time was measured safely under direct colonoscopic visualization. Aspirin prolonged the colon bleeding time. Therefore, endoscopists should be aware of a risk of abnormal bleeding after endoscopic biopsy and polypectomy in patients with aspirin use. Two days were necessary for colon bleeding time to become normalized in patients with aspirin use. PMID- 9407990 TI - Significance of proliferating cell nuclear antigen expression in liver metastasis of colorectal cancer. AB - PURPOSE: The study contained herein was aimed at finding some possible pathologic factors that have significance for the prediction of liver metastasis in colorectal cancer. METHOD: Resected specimens of colorectal cancer from 23 patients with liver metastasis and 30 patients without liver metastasis were subjected to pathologic study, including microscopic characteristics and proliferating cell nuclear antigen immunohistochemistry assay. RESULTS: Strongly positive expression of proliferating cell nuclear antigen was present in 65.21 percent (15/23) of the liver metastasis group, whereas it was found in only 20 percent (6/30) of the group without liver metastasis (P < 0.005). Deeper invasion to the muscularis propria or serosa and less infiltration of lymphocytes surrounding the tumor were more frequently found in the liver metastasis group than in the other group (P < 0.025). CONCLUSION: Extent of proliferating cell nuclear antigen expression, depth of invasion, and reaction of lymphocyte infiltration of the primary tumor could have predictive significance of colorectal cancer in liver metastasis. PMID- 9407991 TI - Carcinoma of the colon. 1954. PMID- 9407992 TI - Anal sphincter reconstruction with a pudendal nerve anastomosis following abdominoperineal resection: report of a case. AB - PURPOSE AND METHODS: We report herein a case of a patient with rectal carcinoma in whom a new anus was constructed following an abdominoperineal resection of the anorectum. This is the first reported case in which reconstruction of the anal sphincter was performed using the lower part of the gluteus maximus muscle with a pudendal nerve anastomosis. The pudendal nerve anastomosis maneuver was designed to achieve proper innervation and function of the external anal sphincter. This newly reconstructed sphincter was physiologically evaluated after surgery. RESULTS: The patient's defunctioning colostomy was not closed following his initial surgery because part of the transposed muscle was devitalized by infection following blood flow damage. However, purposeful contraction of the new sphincter was easy to achieve without special training. The patient's rectal sensation for the desire to defecate was satisfactory. Electromyographic studies demonstrated that the newly reconstructed anal sphincter had characteristics of the original external anal sphincter. CONCLUSIONS: This method is a promising procedure for reconstructing the anal sphincter. PMID- 9407993 TI - Internal hemipelvectomy in the treatment of recurrent carcinoma of the colon: report of a case. AB - PURPOSE: Although extended surgery has been established as an effective method for the treatment of advanced carcinoma of the colon, there are no reports in the literature of en bloc resection of the tumor together with the iliac bone. We report herein a 46-year-old woman with a second local recurrence after right colectomy, with the main objective of showing the possibility of indicating this type of surgery in selected cases. METHODS: In view of the lack of therapeutic options for the case and the absence of metastases, extended curative surgery for recurrent carcinoma of the colon was performed, with en bloc resection of the right iliac bone and of the crural nerve (Type I internal hemipelvectomy). RESULTS: After a 27-month follow-up, the patient is asymptomatic, with no signs of local recurrence or metastases. CONCLUSIONS: In selected cases, recurrent carcinoma of the colon can be treated by extended and aggressive surgery, including bone resection, to obtain an appropriate safety margins. PMID- 9407994 TI - Tumor cell implantation after colonoscopy with biopsies in a patient with rectal cancer: report of a case. AB - PURPOSE: Colonoscopy with biopsy(ies) is performed in patients with colorectal cancer for diagnosis and screening of synchronous lesions. There is some fear of spreading the disease related to implantation of tumor cells at sites of mucosal damage or to hematogenous or lymphatic spread as a result of tumor manipulation. METHODS: A total colonoscopy including biopsies, performed in a patient because of anal blood loss and tenesmus, revealed a circular, polypoid, ulcerated mass from 1.5 to 12 cm above the anal verge. After preoperative radiotherapy, the patient was subjected to surgical intervention. RESULTS: Histopathology revealed a poorly differentiated rectum adenocarcinoma, staged pT4N1Mx. At approximately 12 cm proximally from the tumor, a biopsy taken through a minute irregularity of the mucosa revealed some granulation tissue with adjacent normal colon mucosa. The basis of this granulation tissue, corresponding to submucosa, contained poorly differentiated tumor elements. CONCLUSIONS: This tiny lesion might have been caused by implantation of exfoliated cancer cells in a biopsy site. It may not be without risk to create excessive biopsy lesions in the presence of a manifestly malignant tumor because of the possibility of tumor cell implantation. PMID- 9407995 TI - New pathway for leads in dynamic graciloplasty. AB - Dynamic graciloplasty is one of the methods now used to re-establish anal function. Among complications, pain in the local tissue and skin irritation have been reported, both caused by lead wires. By passing the leads posterior to the adductor longus muscle, the wires do not irritate the local skin and are fixed well without sutures. We performed this procedure in three patients, with good results. The technique is easy, and complications involving the lead wires were minimized. PMID- 9407996 TI - Stimulation of delta1- and delta2-opioid receptors produces amnesia in mice. AB - The effects of intracerebroventricular administration of delta1- and delta2 selective opioid receptor agonists on spontaneous alternation performance, elevated plus-maze behavior and passive avoidance learning including step-down and step-through types were examined in mice. Although the delta1-selective opioid receptor agonist, [D-Pen2,L-Pen5]enkephalin (DPLPE) (1-10 microg) or the delta2-selective opioid receptor agonist, [D-Ala2]deltorphin II (deltorphin) (1 10 microg) did not markedly affect spontaneous alternation performance or elevated plus-maze behavior, DPLPE (1, 3 and/or 10 microg) and deltorphin (3 and 10 microg) inhibited passive avoidance learning including step-down and step through types. The delta1-selective opioid receptor antagonist, 7 benzylidenenaltrexone (3.5 ng), and the delta2-selective opioid receptor antagonist, naltriben (19 ng), significantly antagonized the inhibitory effects of DPLPE (3 microg) and deltorphin (3 microg) on passive avoidance learning, respectively. In contrast, DPLPE (3 microg) or deltorphin (3 microg) did not markedly influence behavioral responses induced by electroshocks during training of passive avoidance learning. Moreover, DPLPE (0.3-3 microg) or deltorphin (0.3 3 microg) failed to significantly affect the radiant heat-induced nociceptive responses. These results suggest that stimulation of delta1- and delta2-opioid receptors produces amnesia, depending on the learning tasks used. PMID- 9407997 TI - Paradoxical behavioral response to apomorphine in tenascin-gene knockout mouse. AB - Tenascin is a large extracellular matrix glycoprotein which is highly expressed in the developing nervous system. To examine the role of tenascin in vivo, we have produced mice in which the tenascin-gene is inactivated. These animals did not easily habituate to unfamiliar circumstances and displayed hyperlocomotion. A dopamine receptor agonist, apomorphine, reduced this hyperlocomotion dose dependently, but this phenomenon was not due to the appearance of apomorphine induced stereotypic behavior, suggesting that tenascin-gene mutant mice have a paradoxical behavioral response to apomorphine compared to wild-type mice. PMID- 9407998 TI - Effect of adenosine receptor agonists and antagonists on morphine-induced catalepsy in mice. AB - Effects of adenosine receptor agonists and antagonists on morphine-induced catalepsy in mice were investigated. The adenosine agonists, NECA (5'-N ethylcarboxamidoadenosine) and S-PIA (S(+)-N6-(2-phenylisopropyl)adenosine) in doses which did not induce any response, increased the cataleptogenic effect produced by morphine. However, the morphine response was decreased and increased by the lower and higher doses of the adenosine receptor agonist, CHA (N6 cyclohexyladenosine), respectively. The adenosine receptor antagonist, theophylline, decreased, but 8-phenyltheophylline increased, the response induced by morphine. Naloxone inhibited the catalepsy induced by morphine or morphine + NECA but not that induced by NECA alone. It is concluded that adenosine A2 receptor activation increases, while adenosine A1 receptor stimulation decreases, the morphine cataleptogenic response. The response to morphine may be mediated through opioid and adenosine receptor mechanisms. PMID- 9407999 TI - A simple probe measures the pharmacokinetics of[125I]RTI-55 in mouse brain in vivo. AB - A simple external radiation detector system was used to assess brain dopamine and serotonin transporters in mice in vivo using [125I]RTI-55. The results were compared to ex vivo dissection data. Methyl 3beta-(4-iodophenyl) tropane-2beta carboxic acid methyl ester (RTI-55 or beta-CIT), a high-affinity cocaine antagonist, binds to presynaptic dopamine and serotonin transporters in the brain. Radiotracer accumulation increased for the first 150 min after intravenous injection and then remained constant. When unlabeled RTI-55 was injected, either before or 60 min after radiotracer administration, a significant decrease in tracer accumulation was observed. [125I]RTI-55 binding was also displaced by blocking doses of 1-[2-[bis(4-fluorophenyl)methoxy]ethyl]-4-[3 phenylpropyl]piperazine dihydrochloride (GBR 12909) and paroxetine. The results were similar to the ex vivo dissection data. The results demonstrate the feasibility of using the probe detector system to study the presynaptic transporter system in vivo in the mouse brain. The technique is applicable to other cerebral neurotransmitter systems. PMID- 9408000 TI - Effect of tyrosine kinase and tyrosine phosphatase inhibitors on aortic contraction and induction of nitric oxide synthase. AB - We studied the effects of the tyrosine kinase inhibitors genistein and tyrphostin and the tyrosine phosphatase inhibitors sodium orthovanadate and phenylarsine oxide on endotoxin-mediated induction of nitric oxide (NO) synthase in rat aorta and its effects on vascular contractility. Genistein (i.p. 10 mg/kg) inhibited the ex vivo vascular hyporesponsiveness to noradrenaline and the aminoguanidine sensitive nitrite accumulation induced by endotoxin (i.p. 5 mg/kg) in aortic rings. Low concentrations of genistein (10(-6) M) and tyrphostin (3 x 10(-6) M) inhibited both endotoxin-induced hyporesponsiveness and nitrite and NOx accumulation in vitro in rat aorta without affecting control nitrite or NOx accumulation or contraction. Higher concentrations of genistein (10(-5) and 5.5 x 10(-5) M), sodium orthovanadate (10(-4) M) and phenylarsine oxide (10(-6) M) produced an irreversible depression of noradrenaline-induced contractions. In the presence of these drugs, endotoxin did not induce further depression of vascular contractility and did not increase nitrite or NOx production. In conclusion, there is a dissociation between the effects of these drugs on vascular smooth muscle contraction and NO synthase induction, the latter being more sensitive to inhibition by these drugs. Surprisingly, tyrosine phosphatase inhibitors produced similar effects to those of tyrosine kinase inhibitors, suggesting that there is a complex relationship between tyrosine kinases and phosphatases in the signalling pathway of agonist-induced vascular smooth muscle contraction and NO synthase induction. PMID- 9408001 TI - PACAP-27 causes negative and positive dromotropic effects in anesthetized dogs. AB - While pituitary adenylate cyclase-activating polypeptide (PACAP) has been identified radioimmunologically in the rat heart, the physiological role of PACAP has not been elucidated in the regulation of the atrioventricular conduction in the heart. We, therefore, determined the dromotropic effects of PACAP-27 injected into the cannulated atrioventricular node artery in the autonomically decentralized heart of the open-chest, anesthetized dog. PACAP-27 caused transient positive followed by negative dromotropic responses in a dose-dependent manner, whereas vasoactive intestinal peptide (VIP) caused only a positive dromotropic response. Atropine and tetrodotoxin blocked the negative dromotropic response to PACAP-27 and after blockade PACAP-27 caused only a positive dromotropic response. Tetrodotoxin and propranolol did not affect the positive dromotropic response to PACAP-27 in atropine-treated dogs. PACAP-27 altered the atrio-His bundle interval but did not alter the His-ventricle interval. These results demonstrate that PACAP-27 prolongs the atrio-His bundle interval due to the liberation of acetylcholine from parasympathetic nerves and decreases it by a non-adrenergic mechanism in the dog heart in situ. PMID- 9408002 TI - The effects of peroxynitrite on rat aorta: interaction with glucose and related substances. AB - Peroxynitrite (1-100 microM) induced a concentration-dependent relaxation of rat aortic rings; the logEC50 and maximum relaxation on endothelium-denuded rings were -5.31 +/- 0.03 and 105 +/- 5%, n = 6, respectively. The presence of the endothelium significantly impaired this relaxation (logEC50, -4.41 +/- 0.04; maximum relaxation, 71 +/- 4%; n = 6); an effect which was reversed by the inhibitor of nitric oxide synthase, N(G)-nitro-L-arginine methyl ester (L-NAME; 100 microM). Incubation with a high concentration of peroxynitrite (1 mM, 10 min followed by washout) had no effect on subsequent relaxation to acetylcholine (0.01-1 microM). It did, however, significantly depress subsequent contraction to phenylephrine (1-300 nM). This depression was dependent upon the presence of D glucose in the Krebs solution, could be reversed by the inhibitor of soluble guanylate cyclase, methylene blue (10 microM) and reversed spontaneously after 2 h. When peroxynitrite (1 mM) was mixed with D-glucose (11 mM) and subsequently neutralised to remove unreacted peroxynitrite, a new more potent relaxant was formed. Despite this, the ability of peroxynitrite (1-100 microM) to produce relaxation of endothelium-denuded rings was similar in normal and glucose-free Krebs. Glycerol (22 mM), which like D-glucose is membrane permeant, also reacted with peroxynitrite (1 mM) to form a new more potent relaxant. L-cysteine (1 mM) had no effect by itself on the tone of aortic rings and when present in the tissue bath had no effect on the ability of peroxynitrite or neutralised peroxynitrite (1-100 microM) to produce relaxation. It did, however, potentiate the relaxant actions of the products formed from the reaction of peroxynitrite with D-glucose or glycerol. The membrane impermeant sugars, mannitol and sorbitol (each 11 mM) also reacted with peroxynitrite (1 mM), but expression of the vasorelaxant properties of their respective derivatives was seen only in the presence of L-cysteine (1 mM). Membrane permeance cannot, however, explain why peroxynitrite reacts with D-glucose and glycerol, but not mannitol or sorbitol to form products with intrinsic relaxant activity, as the product formed from the impermeant sugar, L-glucose (11 mM), also has intrinsic activity. The relaxant potency of this product was equipotent to that formed from D-glucose and was also potentiated by L-cysteine (1 mM). These result confirm that peroxynitrite can react with glucose and other compounds with alcohol functional groups to form vasorelaxant species. The relaxation induced when peroxynitrite is added to rat aortic rings is not, however, dependent upon this reaction since it occurs in glucose-free Krebs. PMID- 9408003 TI - Amelioration by mercaptoethylguanidine of the vascular and energetic failure in haemorrhagic shock in the anesthetised rat. AB - The effects of mercaptoethylguanidine, a dual inhibitor of the inducible nitric oxide (NO) synthase and cyclooxygenase with scavenging effect on peroxynitrite, was studied on the delayed vascular decompensation and cellular energetic failure in a rat model of haemorrhagic shock. Shock was induced by bleeding of the animals to a mean arterial blood pressure of 50 mmHg. At 3 h, animals were resuscitated with Ringers-lactate and monitored for a subsequent 3 h period. In the treated group mercaptoethylguanidine (10 mg/kg/i.v. bolus, followed by 10 mg/kg/i.v. infusion) was administered from the beginning of the resuscitation. Haemorrhagic shock resulted in the upregulation of both the constitutive and the inducible NO synthase, as measured in the lung. In shocked rats mercaptoethylguanidine prevented the increase in plasma nitrite/nitrate and 6 keto-prostaglandin F1alpha levels, ameliorated the decrease in mean arterial blood pressure, and inhibited the development of vascular hyporeactivity of the thoracic aorta ex vivo. A significant nitrotyrosine staining, an indicator of peroxynitrite formation, was found in thoracic aortic rings from shocked animals, which was prevented by mercaptoethylguanidine treatment. In ex vivo experiments in peritoneal macrophages obtained from shocked rats, treatment with mercaptoethylguanidine prevented the reduction in the intracellular NAD+ content, ameliorated the suppression of mitochondrial respiration and reduced the development of DNA single strand breaks. Our data suggest that mercaptoethylguanidine may be an useful tool for the experimental therapy of haemorrhagic shock. PMID- 9408004 TI - Genistein, an inhibitor of tyrosine kinase, prevents the antiarrhythmic effects of preconditioning. AB - The possible involvement of tyrosine kinase in the signal transduction processes associated with the antiarrhythmic effects of ischaemic preconditioning was assessed by the administration, prior to the preconditioning stimulus, of the tyrosine kinase inhibitor genistein (5 mg/kg i.v.) to pentobarbitone anaesthetised male rats. The hearts were then removed, perfused by the Langendorff technique and preconditioned by a single brief (3 min) coronary artery occlusion prior to the 30 min test ischaemic insult. Preconditioning reduced ventricular arrhythmias during occlusion (e.g., premature ventricular beats from 333 +/- 60 in controls to 52 +/- 5 in preconditioned hearts; P < 0.05) but this protective effect was not observed in the hearts of rats given genistein (272 +/- 37 premature beats). A similar abolition of the protective effects of preconditioning could be demonstrated in hearts perfused in vitro with genistein (100 microM). This suggests the involvement of tyrosine kinase activation in this powerful form of endogenous cardioprotection. PMID- 9408005 TI - Pirmenol inhibits muscarinic acetylcholine receptor-operated K+ current in the guinea pig heart. AB - We examined the effects of pirmenol and disopyramide on the muscarinic acetylcholine receptor-operated K+ current (I[K.ACh]) in atrial cells and on experimental atrial fibrillation in isolated guinea-pig hearts. In isolated atrial myocytes, both pirmenol and disopyramide concentration-dependently inhibited the I(K.ACh) induced by carbachol or intracellular loading of GTPgammaS. Their inhibitory effects on the carbachol-induced current were more potent than those on GTPgammaS-induced current, suggesting that these drugs inhibit I(K.ACh) mainly by blocking muscarinic receptors. In Langendorff-perfused hearts these drugs reversed the carbachol-induced decreases in effective refractory periods and atrial fibrillation threshold. These drugs may be useful for the prevention of vagally induced atrial fibrillation. PMID- 9408006 TI - The presence and the effects of neuropeptide Y in rat anococcygeus muscle. AB - Isolated anococcygeus muscle from male rats was examined for the presence of neuropeptide Y-immunoreactive nerves and for the effects of neuropeptide Y on its tone and its contractile/relaxant responses to electrical field stimulation, acetylcholine, guanethidine and noradrenaline. Using peroxidase anti-peroxidase immunohistochemistry in stretch preparation of the anococcygeus, neuropeptide Y immunoreactive nerve fibres were observed, in abundance, running along both vascular as well as non-vascular smooth muscle cells. Neuropeptide Y (> 250 nM) evoked phentolamine and tetrodotoxin-resistant contractile response. Neuropeptide Y, even in subspasmogenic concentrations, potentiated contractions evoked by acetylcholine, guanethidine and noradrenaline. Electrical field stimulation (trains of 3-4 pulses, 0.1 ms, 10 Hz) of the isolated anococcygeus preparation produced robust, phentolamine and tetrodotoxin sensitive contractions. Neuropeptide Y (< 10 nM) exerted a biphasic effect on the electrical field stimulation-evoked contractions; an early potentiation was followed by a delayed and progressive inhibition. Neuropeptide Y (> 10 nM) caused a concentration dependent potentiation of electrical field stimulation-evoked contraction alone, matching its potentiation of noradrenaline-evoked contraction. Electrical field stimulation (5 pulses, 0.1 ms, 10 Hz) of guanethidine (50 microM)-contracted anococcygeus induced a relaxant response and neuropeptide Y (1-100 nM) exerted a concentration-related slight and variable effect on the electrical field stimulation-evoked relaxant response (1 nM, augmentation; 10 nM, no effect; 100 nM, reduction). It is concluded that rat anococcygeus muscle has a rich neuropeptide Y-containing innervation and neuropeptide Y is mostly stored within adrenergic nerves. The main functional roles of neuropeptide Y in the anococcygeus muscle are likely to be post-junctionally mediated facilitation and prejunctionally mediated inhibition of adrenergic motor transmission. PMID- 9408007 TI - Characterization of tolerance to the anti-leakage effect of formoterol in rat airways. AB - These experiments addressed the question of whether the anti-plasma leakage action of beta2-adrenoceptor agonists in rat airways is subject to tolerance. Pathogen-free F344 rats were pretreated with the highly selective, long-acting beta2-adrenoceptor agonist, formoterol (0, 0.1, 1, 10 microg/kg, i.p.) for 7 days; 24 h later the effectiveness of acute doses of formoterol (0, 0.1, 1, 10 microg/kg, i.v.) was tested against substance P-induced plasma leakage. The anti leakage effect of formoterol was not subject to tolerance with the low or intermediate pretreatment dose. Pretreatment with 10 microg/kg formoterol reduced the effectiveness of the 1 microg/kg acute dose but not the 10 microg/kg acute dose. We conclude that tolerance to the anti-leakage effect of formoterol can occur, but airway vessels are likely to retain functionally coupled receptors even after high dose pretreatment. PMID- 9408008 TI - Pharmacological characterisation and autoradiographic localisation of a putative dopamine D3 receptor in the rat kidney. AB - The pharmacological profile and the microanatomical localisation of a putative dopamine D3 receptor in the rat renal cortex were investigated using radioligand binding assay and light microscope autoradiography techniques. [3H]7-hydroxy-N,N di-n-propyl-2-aminotetraline ([3H]7-OH-DPAT) was used as a ligand. [3H]7-OH-DPAT was bound specifically to sections of renal cortex. The binding was time-, temperature- and concentration-dependent, of high affinity and guanine nucleotide insensitive. The dissociation constant (Kd) value was 0.57 +/- 0.02 nM and the maximum density of binding sites (Bmax) was 62.4 +/- 3.5 fmol/mg tissue. The pharmacological profile of [3H]7-OH-DPAT binding to sections of rat renal cortex suggests the labelling of a dopamine D3 receptor. Light microscope autoradiography revealed the accumulation of the radioligand primarily within cortical tubules and to a lesser extent in the glomerular tuft. In glomeruli, binding sites were found mainly in mesangium and mesangial cells. The demonstration of a putative dopamine D3 receptor in slide-mounted sections of rat renal cortex suggests that appropriate radioligand binding assay techniques combined with autoradiography, may contribute to characterise peripheral dopamine receptor subtypes. PMID- 9408009 TI - Serotonin inhibits nitric oxide synthesis in rat vascular smooth muscle cells stimulated with interleukin-1. AB - We investigated the effects of serotonin (5-hydroxytryptamine; 5-HT) on nitric oxide (NO) synthesis in vascular smooth muscle cells. We measured the production of nitrite, a stable metabolite of NO, and the expression of inducible NO synthase protein in cultured rat vascular smooth muscle cells. Incubation of the cultures with interleukin-1beta (10 ng/ml) caused a significant increase in nitrite production. 5-HT inhibited nitrite production by interleukin-1beta stimulated vascular smooth muscle cells in a concentration-dependent manner (10( 8)-10(-5) M). 5-HT-induced inhibition of nitrite production was accompanied by decreased inducible NO synthase protein accumulation in vascular smooth muscle cells. Addition of the 5-HT2 receptor antagonist ketanserin, but not the 5-HT1A receptor antagonist spiroxatrine, inhibited the effect of 5-HT. On the other hand, the 5-HT2 receptor agonist alpha-methyl-5-HT, but not the 5-HT1A receptor agonist (+/-)-8-hydroxy-2-(di-n-propylamino) tetralin, decreased interleukin 1beta-induced nitrite production by vascular smooth muscle cells. 5-HT significantly increased protein kinase C activity in vascular smooth muscle cells, and the protein kinase C inhibitor calphostin C dose-dependently abolished the effect of 5-HT on nitrite production. After protein kinase C activity was functionally depleted by treatment of cells with phorbol 12-myristate 13-acetate for 24 h, the effect of 5-HT was abolished. These results indicate that 5-HT acts on 5-HT2 receptors and inhibits NO synthesis in interleukin-1beta-stimulated vascular smooth muscle cells at least partially through a protein kinase C dependent pathway. PMID- 9408010 TI - Bidirectional trafficking of membrane glycoproteins following platelet activation in suspension. PMID- 9408011 TI - Occult cancer in patients with venous thromboembolism: which patients, which cancers. AB - We have previously demonstrated that patients with idiopathic venous thromboembolism (VTE) have a higher frequency of underlying cancer. Now we present a retrospective analysis of our 5-year experience with a series of 674 consecutive otherwise healthy patients, and a more restricted battery of diagnostic tests. Occult cancer was found in 15 patients during admission. The diagnostic tools which led to suspect occult cancer were: abdominal CT-scan (4 patients); high carcinoembryonic levels (2 patients); and high prostate-specific antigen levels (9 patients). Eight further patients were diagnosed of cancer after discharge. Cancer was more commonly found in patients with idiopathic VTE: 13/105 patients (12%) versus 10/569 patients (2%); p <0.01; O.R.: 7.9 (95% CI: 3.14-20.09). During the same period of time we diagnosed VTE in 147 patients with previously known cancer. When overall considered, VTE was the first sign of malignancy in most patients with prostatic and pancreatic carcinoma. On the contrary, most patients with breast, lung, uterine and brain cancers developed VTE as a terminal event of the disease. At variance with VTE patients and previously known cancer, most patients with occult malignancy were at an early stage. Further studies are needed to confirm whether patients with idiopathic VTE could benefit from screening for occult cancer. Meanwhile, our findings may serve as guidelines for physicians in this field. PMID- 9408012 TI - Testing for occult cancer in patients with idiopathic deep vein thrombosis--a decision analysis. AB - OBJECTIVE: To determine the effectiveness and cost-effectiveness of testing for occult cancer in idiopathic deep vein thrombosis (IDVT). DESIGN: Threshold analysis was performed on the risk-adjusted cancer prevalence in a cost effectiveness model of ideal testing for selecting cancers with potentially desirable utility (candidate cancers). Decision analysis was employed to compare different testing programs for candidate cancers with that of no testing. Life expectancy (LE) of early- and late-detected cancers and costs of testing were the dimensions of utility. Cost-effectiveness was expressed as marginal cost per year of life saved. The perspective of the third payer was adopted, and a discount rate of 3% was applied to both costs and benefits. DATA SOURCES: Risk of cancer in IDVT, testing policies, test characteristics, and LE were gathered from literature. Costs were provided from our hospital rate book and accounting service. RESULTS: Ideal testing would support a gain of LE of 40 days or more for prostate, colon and bladder cancer in males and for colon, breast and endometrium cancer in females aged from 60 to 69 years. Testing females with colonoscopy and mammography in any sequence provides 70 days of life gained. Testing males with colonoscopy provides 27 days of life gained. Lower and older ages reduce testing effectiveness. The qualitative results are stable over plausible ranges of test characteristics, while variations in the value of benefit for early cancer diagnosis may modify the strategy. Incremental cost-effectiveness ranges from $1,789 to $ 6,979 per year of life gained. CONCLUSIONS: According to the effectiveness criterion adopted, the only worthwhile investigation strategy includes colon and breast cancer in females. Testing for colon cancer in males is desirable at a lower criterion of effectiveness. All the strategies are cost effective. PMID- 9408013 TI - Levels of prothrombin fragment F1+2 in patients with hyperhomocysteinemia and a history of venous thromboembolism. AB - Increased thrombin generation occurs in many individuals with inherited defects in the antithrombin or protein C anticoagulant pathways and is also seen in patients with thrombosis without a defined clotting abnormality. Hyperhomocysteinemia (H-HC) is an important risk factor of venous thromboembolism (VTE). We prospectively followed 48 patients with H-HC (median age 62 years, range 26-83; 18 males) and 183 patients (median age 50 years, range 18-85; 83 males) without H-HC for a period of up to one year. Prothrombin fragment F1+2 (F1+2) was determined in the patient's plasma as a measure of thrombin generation during and at several time points after discontinuation of secondary thromboprophylaxis with oral anticoagulants. While on anticoagulants, patients with H-HC had significantly higher F1+2 levels than patients without H-HC (mean 0.52 +/- 0.49 nmol/l, median 0.4, range 0.2-2.8, versus 0.36 +/- 0.2 nmol/l, median 0.3, range 0.1-2.1; p = 0.02). Three weeks and 3, 6, 9 and 12 months after discontinuation of oral anticoagulants, up to 20% of the patients with H-HC and 5 to 6% without H-HC had higher F1+2 levels than a corresponding age- and sex matched control group. 16% of the patients with H-HC and 4% of the patients without H-HC had either F1+2 levels above the upper limit of normal controls at least at 2 occasions or (an) elevated F1+2 level(s) followed by recurrent VTE. No statistical significant difference in the F1+2 levels was seen between patients with and without H-HC. We conclude that a permanent hemostatic system activation is detectable in a proportion of patients with H-HC after discontinuation of oral anticoagulant therapy following VTE. Furthermore, secondary thromboprophylaxis with conventional doses of oral anticoagulants may not be sufficient to suppress hemostatic system activation in patients with H-HC. PMID- 9408015 TI - Repeated exercise induces release of soluble P-selectin in patients with intermittent claudication. AB - Controversy exists as to whether exercise in patients with intermittent claudication causes a harmful biochemical effect associated with an ischaemia reperfusion injury of skeletal muscle. We report on exercise-induced changes in neutrophil activation, soluble P-selectin and von Willebrand factor in 34 patients with intermittent claudication and 12 matched controls. Von Willebrand factor (vWF) showed a cyclical pattern of response to exercise in control subjects (rising from 103 +/- 8 to 119 +/- 7 U/dl); claudicants did not show this pattern but had higher levels of vWF throughout (p <0.03). There was no consistent pattern of response in neutrophil hydrogen peroxide production to exercise in either claudicants or control subjects. Soluble P-selectin levels increased after exercise, but this only reached statistical significance after repeated exercise in claudicants (rising from 320 +/- 28 to 357 +/- 28 ng/ml). This rise in soluble P-selectin after exercise may indicate progressive platelet activation which may contribute to the excess cardiovascular mortality that claudicants are prone to. PMID- 9408014 TI - Endothelial marker proteins in hyperhomocysteinemia. AB - Hyperhomocysteinemia is associated with severe, premature atherosclerosis and thromboembolism. The mechanisms involved in the atherogenic and thrombotic complications of hyperhomocysteinemia are not understood. It has been suggested that hyperhomocysteinemia predisposes to atherosclerosis by injuring the vascular endothelium. Whether hyperhomocysteinemia is independently associated with changed endothelial function, either in the absence or the presence of clinically manifest atherosclerotic disease, is, however, not known. Therefore we investigated, both in patients with peripheral arterial occlusive disease and in healthy individuals, whether plasma protein markers of endothelial function differed between subjects with, and subjects without hyperhomocysteinemia. We studied 80 individuals under the age of 56 years: healthy individuals with (n = 20) and without (n = 20) hyperhomocysteinemia and patients with peripheral arterial occlusive disease with (n = 20) and without (n = 20) hyperhomocysteinemia. The following endothelium-derived proteins were measured as markers of endothelial cell function: von Willebrand factor (vWf) and von Willebrand factor propeptide (vWf: AgII), tissue-type plasminogen activator (tPA), plasminogen activator inhibitor-1 (PAI-1), cellular fibronectin (cFN) and thrombomodulin (TM). In addition we assessed C-reactive protein (CRP). vWf, vWf: AgII, tPA and CRP were significantly higher in the patients with peripheral arterial occlusive disease than in the healthy individuals. No differences in marker protein plasma levels were found between individuals with, and those without hyperhomocysteinemia, apart from vWf, which was significantly raised in hyperhomocysteinemic as compared to normohomocysteinemic patients. We did not find any evidence for an independent association between hyperhomocysteinemia and protein markers of endothelial cell function in healthy subjects. PMID- 9408016 TI - Free protein S deficiency is a risk factor for venous thrombosis. AB - A recent study suggests that protein S deficiency is not a risk factor for venous thrombosis. Since this unexpected finding would have important clinical implications if confirmed, we performed a case-control study with the aim to determine the prevalence of protein S deficiency in patients with thrombosis and in healthy individuals taken from the general population and the relative risk of thrombosis in protein S-deficient patients. Free protein S concentration was measured in 327 consecutive patients with at least one venous thrombotic episode and in 317 age- and sex-matched control individuals. Different normal reference ranges were obtained and adopted for men and women. Protein S deficiency was found in 3.1% (95% CI: 1.5-5.2) of patients and in 1.3% of controls (95% CI: 0.3 2.8). Ten patients and 4 control subjects had protein S deficiency, which determined a relative risk of thrombosis (sex- and age-adjusted odds ratio) of 2.4 (95% CI: 0.8-7.9). When men and women were analyzed separately, the risk was 5.0 (95% CI: 0.6-43.6) and 1.6 (95% CI: 0.4-6.7) respectively. PS-deficient men had more thrombotic episodes than women and later in life. Multivariate analysis established that sex was an independent determinant of the number of episodes, as was age, while PS deficiency was not. However sex and PS deficiency status were both determinants of age at first thrombotic episode. PMID- 9408017 TI - Haemophilia B in female twins caused by a point mutation in one factor IX gene and nonrandom inactivation patterns of the X-chromosomes. AB - Haemophilia B is a X-linked recessive bleeding disorder with an incidence of 1:25,000-30,000 male birth. Usually female carriers are clinically normal. Phenotypic expression of the disease in female carriers is extremely rare. We describe cytogenetically inconspicuous female identical twins both with factor IX levels below 2%, prolonged bleeding after venipuncture as well as haematomas after intramuscular injections. The father, suffering from a severe haemophilia B, is deceased. By sequencing one point mutation was characterized in heterozygote condition in the factor IX gene of the probands at nt 17678. This mutation leads to the substitution cystein 88 to tyrosine in the growth factor domain of the factor IX. Investigation of the X-chromosomal inactivation by comparison of methylation patterns of genomic DNA at locus DXS255 after digestion with Pst I and Pst I +Hha I and hybridisation with the probe M27beta indicated a nonrandom pattern of X-chromosomal inactivation in the twins. In both girls, only the paternal X-chromosome was the active one leading to the phenotypic expression of haemophilia in the female carriers. PMID- 9408018 TI - A multicenter pharmacosurveillance study for the evaluation of the efficacy and safety of recombinant factor VIII in the treatment of patients with hemophilia A. German Kogenate Study Group. AB - In this open multicenter study the safety and efficacy of recombinant factor VIII (rFVIII) was assessed in 39 previously treated patients with hemophilia A (factor VIII basal activity < or = 15%). Recombinant FVIII was administered for prophylaxis and treatment of bleeding episodes and for surgical procedures. A total of 3679 infusions of rFVIII were given. Efficacy of rFVIII as assessed by subjective evaluation of response to infusion and mean annual consumption of rFVIII was comparable to that of plasma derived FVIII concentrates. The incremental recovery of FVIII (2.4 +/- 0.83%/IU/kg, 2.12 +/- 0.61%/IU/kg, resp.) was within the expected range. No clinical significant FVIII inhibitor was detected in this trial. Five of 16 susceptible patients showed a seroconversion for parvovirus B19. However, the results are ambiguous in two cases and might be explained otherwise in one further case. Thus, in two patients a reliable seroconversion for parvovirus B19 was observed. PMID- 9408019 TI - Factor V Leiden and thermolabile methylenetetrahydrofolate reductase in extreme old age. AB - Both factor V Leiden and the C677T methylenetetrahydrofolate reductase (MTHFR) gene mutation are associated with premature vascular disease, and yet are found at surprisingly high allele frequencies in European populations, 2.7% and 35% respectively. We have investigated the prevalence of these mutations in 87 UK residents over the age of ninety, to look for any evidence of their association with premature death. Five factor V Leiden heterozygotes were found, giving an allele frequency of 2.9%, similar to that in the general UK population. The frequency of the thermolabile C677T MTHFR mutation was 36% with 11% homozygotes, again similar to that in the UK population; these genotypes are in Hardy-Weinberg equilibrium, suggesting that there is not strong selection against the homozygous state. One person was both heterozygous for factor V Leiden and homozygous for the C677T mutation. This study suggests that neither factor V Leiden nor thermolabile MTHFR are risk factors for premature death. PMID- 9408020 TI - Proposal for objective evaluation of the performance of various functional APC resistance tests in genotyped patients. AB - The aim of the present study was to evaluate the relative performance of five screening methods for APC resistance caused by the factor V:Q506 mutation: the original method Coatest APC Resistance Chromogenix, a modified method using the same reagents but a predilution 1+4 of the plasma in a factor V deficient plasma from Stago (Stago deficient V) or from Chromogenix (V-DEF Plasma), the Coatest APC Resistance V (Chromogenix), and Accelerimat from bioMerieux. Normalization was done against a pool of normal plasmas for the methods from Chromogenix. The study included 350 subjects, 219 were genotyped (174 FV:R506R, 42 FV:Q506R, 3 FV:Q506Q) and most of them were assessed by more than one method. Uncertainty in predicting the FV genotype was evaluated by statistical analysis, which provided a way to quantitate the performance of the different diagnostic approaches. Performance of each test was evaluated by its sensitivity, specificity, R.O.C. curves, positive and negative likelihood ratios (LR), and the overall performance was determined by two parameters derived from the LR curves : the maximum LR value obtained at the crossover of the two curves, and the distance between the two curves for LR = 10. Coatest APC Resistance V and Accelerimat were proven to be the methods most able to discriminate for factor V:Q506, while normalization was not shown to improve the screening performance. The original method from Chromogenix was confirmed to undergo many influences (factor XII, PAI-1, thrombin antithrombin complexes, antithrombin III, hematocrit). Although a very good improvement was provided by the newest methods, they were shown to be influenced by protein S and/or factor V levels in the sample plasma. PMID- 9408021 TI - Antithrombotic and hemorrhagic effects of DX-9065a, a direct and selective factor Xa inhibitor: comparison with a direct thrombin inhibitor and antithrombin III dependent anticoagulants. AB - (+)-2S-2-[4-[[(3S)-1-Acetimidoyl-3- pyrrolidinyl]oxy]phenyl]-3-[7-amidino-2 naphthyl]propanoic acid hydrochloride pentahydrate (DX-9065a) is an antithrombin III (AT III)-independent and selective inhibitor of activated blood coagulation factor X (FXa). The aim of the present study was to compare the antithrombotic and hemorrhagic effects of DX-9065a with a direct thrombin inhibitor and AT III dependent anticoagulants in rat models of thrombosis and bleeding. Rats were administered intravenously DX-9065a (0.1-1 mg/kg/h), argatroban (0.1-1 mg/k/h), low molecular weight heparin (25-100 anti-XaU/kg/h), unfractionated heparin (25 100 anti-XaU/kg/h) or Orgaran (30-300 anti-XaU/kg/h) for 1 h. DX-9065a dose dependently inhibited both thrombus formation and elevation in plasma thrombin-AT III complex (TAT) level in a copper wire-inserted arteriovenous (AV) shunt model in rats. The dose required for 50% inhibition of thrombus formation was 0.27 mg/kg/h. DX-9065a did not prolong transection bleeding time up to 7.78 mg/kg/h. Argatroban and AT III-dependent anticoagulants also inhibited both thrombus formation and TAT elevation, but prolonged bleeding time at a slightly higher dose than the effective dose. These results suggest that direct and selective inhibition of factor Xa by DX-9065a is preferable for the treatment of thrombosis in the aspect of lack of compromising primary hemostasis. PMID- 9408022 TI - Mechanism of inhibitory action on platelet activation of a phospholipase A2 isolated from Lachesis muta (Bushmaster) snake venom. AB - Crude venom from Lachesis muta exhibited procoagulant, proteolytic and phospholipase A2 activities. A phospholipase A2, denoted LM-PLA2 was purified from L. muta venom to homogeneity, through a combination of chromatographic steps involving gel-filtration on Sephacryl S-200 HR and reverse phase chromatography on a C2/C18 column. LM-PLA2 presented a single polypeptide chain with an isoelectric point at pH 4.7 and apparent molecular weight of 17 kDa. Partial aminoacid sequence indicated a high degree of homology for LM-PLA2 with other PLA2 from different sources. LM-PLA2 displayed a potent enzymatic activity as measured by indirect hemolysis of red blood cells but it was neither lethal when injected i.p. into mice nor did it present anticoagulant activity. Furthermore, LM-PLA2 displayed a moderate inhibitory activity on the aggregation of rabbit platelets induced by low levels of ADP, thrombin and arachidonate. In contrast, platelet aggregation induced by high doses of collagen was strongly inhibited by LM-PLA2 as well as ATP-release. Treatment of the protein with p-bromophenacyl bromide or 2-mercaptoethanol, as well as thermal inactivation studies, suggested that the platelet inhibitory effect of LM-PLA2 is dependent on its enzymatic activity. Thus, the platelet inhibitory activity of LM-PLA2 was shown to be dependent on the hydrolysis of plasma phospholipids and/or lipoproteins, most probably those rich in phosphatidylcholine. Surprisingly, lysophosphatidylcholine released by LM-PLA2 from plasma was shown to preferentially inhibited collagen induced platelet aggregation, in contrast to other PLA2s, whose plasma hydrolytic products indistinctly affect platelet's response to several agonists. PMID- 9408023 TI - Enhancement by ticlopidine of the inhibitory effect on in vitro platelet aggregation of the glycoprotein IIb/IIIa inhibitor tirofiban. AB - We determined the effects of combining the glycoprotein IIb/IIIa inhibitor tirofiban (MK-383, Aggrastat) and ticlopidine on inhibition of ADP-induced platelet aggregation, inhibition of collagen-induced platelet aggregation, and prolongation of bleeding time in 5 healthy male volunteers. The study consisted of 2 periods. In period 1, tirofiban was administered intravenously as a bolus injection at a dose of 5.0 microg/kg for 5 min, then as a continuous infusion at a rate of 0.05 microg/kg/min for 5 h 55 min. In period 2, ticlopidine was given orally at a dose of 200 mg once daily for 4 days, after which tirofiban was administered in the same manner as in period 1, starting 2 h after the last dose of ticlopidine. The percent inhibition of platelet aggregation induced by ADP 5 microM at the end of tirofiban infusion in periods 1 and 2 was 73.6 +/- 2.6 and 87.1 +/- 5.7% (mean +/- SD), respectively. The corresponding values for inhibition of platelet aggregation induced by collagen 2 microg/ml were 45.4 +/- 36.1 and 82.8 +/- 27.0%, respectively. In contrast, the percent inhibition of platelet aggregation induced by ADP and collagen after treatment with ticlopidine alone was 15.8 +/- 20.2 and 2.2 +/- 4.8%, respectively. Compared with tirofiban alone, the combination of tirofiban and ticlopidine significantly enhanced inhibition of platelet aggregation induced by both ADP and collagen (p <0.02 and p <0.012, respectively). The inhibitory effects of ticlopidine alone were not statistically significant. Tirofiban prolonged bleeding time both in period 1 and in period 2. However, tirofiban and ticlopidine combined did not affect bleeding time. Ticlopidine administration did not alter the pharmacokinetics of tirofiban. In conclusion, at the doses of tirofiban and ticlopidine used in this study, the combination of the two drugs enhanced inhibition of platelet aggregation but did not prolong bleeding time, suggesting that appropriate doses of tirofiban can be used concomitantly with ticlopidine with no further safety concerns but with potential additional clinical effects. PMID- 9408024 TI - Antivasoconstrictor and antiaggregatory activities of picotamide unrelated to thromboxane A2 antagonism. AB - Picotamide is a dual thromboxane (Tx) A2 receptor antagonist/Tx synthase inhibitor although some observations suggest an anti-vasoconstrictor effect independent of TxA2 inhibition/antagonism. The aim of our study was to assess whether picotamide antagonises vascular contractions induced by different vasoactive substances in vitro. Picotamide inhibited competitively the contraction of rabbit aortic rings induced by the TxA2 mimetic U46619 (pA2 = 3.59) but also the contractions induced by phenylephrine (pA2 = 3.93) and serotonin (5-HT) (pA2 = 5.81) although in a not competitive way. Picotamide did not inhibit potassium-induced contractions, thus excluding aspecific effects on vascular smooth muscle. Picotamide inhibited 5-HT-induced platelet aggregation in vitro with an IC50 (212 microM) similar to that found when other aggregating stimuli are used, but it did not affect shape change (IC50 > 1 mM) suggesting that the effects of picotamide can not be ascribed to 5-HT2-receptor antagonism; in the same experimental conditions neither a Tx-receptor antagonist (BM13.177) nor a dual Tx-receptor antagonist/synthase inhibitor (ridogrel) affected 5-HT induced platelet responses. Our studies demonstrate that picotamide exerts antivasoconstrictor and platelet inhibitory effects unrelated to TxA2 antagonism. This activity may contribute to the anti-thrombotic/anti-ischaemic effects of the drug in vivo. PMID- 9408025 TI - Naturally produced extracellular matrix is an excellent substrate for canine endothelial cell proliferation and resistance to shear stress on PTFE vascular grafts. AB - PURPOSE: Successful development of a vascular prosthesis lined with endothelial cells (EC) may depend on the ability of the attached cells to resist shear forces after implantation. The present study was designed to investigate EC detachment from extracellular matrix (ECM) precoated vascular prostheses, caused by shear stress in vitro and to test the performance of these grafts in vivo. METHODS: Bovine aortic endothelial cells were seeded inside untreated polytetrafluoro ethylene (PTFE) vascular graft (10 x 0.6 cm), PTFE graft precoated with fibronectin (FN), or PTFE precoated with FN and a naturally produced ECM (10(6) cells/graft). Sixteen hours after seeding the medium was replaced and unattached cells counted. The strength of endothelial cell attachment was evaluated by subjecting the grafts to a physiologic shear stress of 15 dynes/cm2 for 1 h. The detached cells were collected and quantitated. PTFE or EC preseeded ECM coated grafts were implanted in the common carotid arteries of dogs. RESULTS: While little or no differences were found in the extent of endothelial cell attachment to the various grafts (79%, 87% and 94% of the cells attached to PTFE, FN precoated PTFE, or FN+ECM precoated PTFE, respectively), the number of cells retained after a shear stress was significantly increased on ECM coated PTFE (20%, 54% and 85% on PTFE, FN coated PTFE, and FN+ECM coated PTFE, respectively, p <0.01). Implantation experiments in dogs revealed a significant increase in EC coverage and a reduced incidence of thrombus formation on ECM coated grafts that were seeded with autologous saphenous vein endothelial cells prior to implantation. CONCLUSION: ECM coating significantly increased the strength of endothelial cell attachment to vascular prostheses subjected to shear stress. The presence of adhesive macromolecules and potent endothelial cell growth promoting factors may render the ECM a promising substrate for vascular prostheses. PMID- 9408026 TI - Trypsin- and SLIGRL-induced vascular relaxation and the inhibition by benzamidine derivatives. AB - Serine proteinases are involved in several physiological processes and elicit profound cellular effects in a variety of tissues. Besides the thrombin receptor a second receptor, activated by trypsin, the proteinase-activated receptor 2 (PAR 2), was cloned and characterized. Both enzymes generate a new extracellular N terminus by limited proteolytic cleavage which functions as tethered ligand to activate the receptor. Synthetic peptides corresponding to the sequences of the newly generated N-terminus are able to mimic the effects of the enzymes. In porcine pulmonary arteries trypsin and the receptor-derived peptide SLIGRL elicited an endothelium-dependent transient relaxation of PGF2alpha-precontracted vessels. The EC50 values for trypsin and SLIGRL amounted to 1.1 +/- 0.2 nM and 5.4 +/- 0.6 microM, respectively. Trypsin and SLIGRL caused a homologous desensitization but thrombin and the thrombin receptor-activating peptide SFLLRN were still able to elicit pronounced relaxant effects. The trypsin- and SLIGRL induced relaxant responses were markedly diminished after blockade of the nitric oxide synthesis by N(G)-nitro-L-arginine methyl ester (200 microM) and were absent in endothelium-denuded vessels. Indomethacin and hirudin did not influence the relaxant effects. The effect of trypsin but not that of SLIGRL was blocked by the proteinase inhibitor aprotinin suggesting that only proteolytically active trypsin activates the receptor. Benzamidine derivatives of the 3 amidinophenylalanine type with different affinity for trypsin and thrombin inhibited the vascular effects of trypsin (IC50 0.007-0.7 microM) correlating with its antitrypsin activity. The data suggest that the vascular effects of trypsin and SLIGRL are mediated through activation of PAR-2 which differs from the thrombin receptor. PMID- 9408027 TI - Antithrombotic effect of two low molecular weight thrombin inhibitors and a low molecular weight heparin in a caval vein thrombosis model in the rat. AB - A sensitive thrombosis model with a high reproducibility was developed in the rat, utilizing stasis of the caval vein and a standardized surgical trauma as the only thrombogenic stimuli. Since no procoagulant substances were used, the results of the present study might be relevant in a clinical situation. The antithrombotic effect of two recently synthesized low-molecular-weight thrombin inhibitors have been compared to dalteparin, (Fragmin) a low-molecular-weight heparin fragment. Each compound was studied at 8 different doses with 10 rats in each group. On a gravimetric basis, the thrombin inhibitor melagatran was twice as potent as dalteparin (ED50 16 and 33 microg/kg per h, respectively). The second thrombin inhibitor, inogatran, had an intermediate effect, with an ED50 of 24 microg/kg per h. No differences in antithrombotic effect were, however, found when the compounds were compared at anticoagulant equivalent doses (same APTT prolongation). A 50% reduction in the mean thrombus weight was obtained when APTT was prolonged to 1.2 to 1.3 times the pretreatment value. PMID- 9408029 TI - Lack of association between thrombosis in primary antiphospholipid syndrome and the recently described thrombophilic 3'-untranslated prothrombin gene polymorphism. PMID- 9408028 TI - A new model for quantitative in vivo microscopic analysis of thrombus formation and vascular recanalisation: the ear of the hairless (hr/hr) mouse. AB - The alteration of rheological blood properties as well as deterioration of vascular perfusion conditions and cell-cell interactions are major determinants of thrombus formation. Herein, we present an experimental model which allows for quantitative in vivo microscopic analysis of these determinants during both thrombus formation and vascular recanalisation. The model does not require surgical preparation procedures, and enables for repeated analysis of identical microvessels over time periods of days or months, respectively. After i.v. administration of FITC-dextran thrombus formation was induced photochemically by light exposure to individual arterioles and venules of the ear of ten anaesthetised hairless mice. In venules, epi-illumination induced rapid thrombus formation with first platelet deposition after 0.59 +/- 0.04 min and complete vessel occlusion within 7.48 +/- 1.31 min. After a 24-h time period, 75% of the thrombosed venules were found recanalised. Marked leukocyte-endothelial cell interaction in those venules indicated persistent endothelial cell activation and/or injury, even after an observation period of 7 days. In arterioles, epi illumination provoked vasomotion, while thrombus formation was significantly (p <0.05) delayed with first platelet deposition after 2.32 +/- 0.22 min and complete vessel occlusion within 20.07 +/- 3.84 min. Strikingly, only one of the investigated arterioles was found recanalised after 24 h, which, however, did not show leukocyte-endothelial cell interaction. Heparin (300 U/kg, i.v.) effectively counteracted the process of thrombus formation in this model, including both first platelet deposition and vessel occlusion. We conclude that the model of the ear of the hairless mouse allows for distinct in vivo analysis of arteriolar and venular thrombus formation/recanalisation, and, thus, represents an interesting tool for the study of novel antithrombotic and thrombolytic strategies, respectively. PMID- 9408030 TI - Acquired protein S deficiency, likely due to anti-PS autoantibodies, following a thrombotic event in a patient with a systemic lupus erythematosus. PMID- 9408031 TI - Ratiometric measurements with fluorescent indicators should be interpreted cautiously in studies of platelet calcium signaling for von Willebrand factor. PMID- 9408032 TI - A novel polymorphism in exon 10 of the factor V gene inducing an amino acid replacement of arginine to lysine at codon 485. PMID- 9408033 TI - Serum thrombopoietin levels following orthotopic liver transplantation. PMID- 9408034 TI - Integration of local inputs in visual cortex. AB - In mammalian visual cortex, local connections are ubiquitous, extensively linking adjacent neurons of all types. In this study, optical maps of intrinsic signals and responses from single neurons were obtained from the same region of cat visual cortex while the effectiveness of the local cortical circuitry was altered by focally disinhibiting neurons within a column of known orientation preference. Maps of intrinsic signals indicated that local connections provide strong and functional subthreshold inputs to neighboring columns of other orientation preferences, altering the observed orientation preference to that of the disinhibited column. However, measuring the suprathreshold response using single cell recordings revealed only mild changes of preferred orientation over the affected region. Because strongly tuned subthreshold inputs from cortex only marginally affect the tuning of a cortical cell's output, it is concluded that local cortical inputs are integrated weakly compared to geniculate inputs. Such circuitry potentially allows for the normalization of responses across a wide range of input activity through local averaging. PMID- 9408035 TI - Differential expression of D1 and D5 dopamine receptors in the fetal primate cerebral wall. AB - Previous film autoradiographic studies demonstrated that, during corticogenesis, dopamine receptors of the D1 class are abundant in the embryonic primate cerebral wall. In the present study, we expand these findings by identifying the cellular elements of the fetal occipital cerebral wall expressing D1 and D5 subtypes of the D1 dopamine receptor class. We have examined tissue from monkey fetuses collected at 70, 90 and 120 days of gestation using antibodies directed against C termini of the D1 and D5 dopamine receptors. At all three embryonic ages studied, we found D1 and D5 receptors expressed by multiple cell types of the embryonic cerebral wall. Both D1 and D5 receptor proteins are produced by pyramidal neurons of the cortical plate and by a variety of interstitial neurons of the subplate and intermediate zones. D1 and D5 receptors are also present in cells of the proliferative ventricular and subventricular zones, some of which were identified as dividing cells. In addition, D1 and D5 receptors are detectable in the protoplasmic astroglial and ependymal cells distinguishable in monkey fetuses collected at 120 days of gestation. Some cellular elements of the embryonic monkey cerebral wall express only one subtype of the D1 dopamine receptor class. For example, embryonic Cajal-Retzius neurons in the marginal zone and migrating neurons in the intermediate zone are immunoreactive only to D5 antisera. In contrast, radial glia can be labeled only with D1 receptor-specific antisera. Finally, only D1 receptors are detectable in the blood vessels penetrating the embryonic monkey cerebral wall. Based on these observations, we propose that dopamine receptors of the D1 class play an important role in regulating cerebral cortical formation and that D1 and D5 receptor subtypes may participate in regulation of different aspects of this process. PMID- 9408036 TI - The entorhinal cortex: an examination of cyto- and myeloarchitectonic organization in humans. AB - The entorhinal cortex (ERC) has been implicated in the pathophysiology of Alzheimer's disease, schizophrenia and other disorders affecting cognitive functions. While powerful anatomical and histochemical methods (immunohistochemistry, in situ hybridization, etc.) may be applied (although with limitations) to postmortem human brain, each analysis should utilize a cytoarchitectonic approach to provide appropriate comparisons within the subdivisions of the ERC. Accordingly, we describe here the normal cyto- and myeloarchitecture of the human ERC as a prerequisite for the accompanying study of this region in schizophrenia. Our parcellation of this cortex differs from previous treatments in three ways. First, we adopted specific criteria of inclusion to define each subdivision of the region. Although distinctive ERC features are most prominent in the intermediate portion of this region, at least one of these features was considered the minimum necessary criterion to include adjacent tissue in the entorhinal area. Second, we used morphometric measurements (neuronal size and density as well as subdivisional volume and laminar thickness) to support our qualitative evaluation. Third, we have applied to the human ERC the conventional cytoarchitectonic nomenclature of the entorhinal cortex used previously in studies of non-human primates. This allows a more accurate extrapolation of the available numerous experimental anatomical, physiological and psychological data on this region to the human. As in the monkey, the five main subareas were recognized in the human (prorhinal, lateral, intermediate, sulcal and medial) but three required further subdivision (intermediate, sulcal and medial). The morphometric results obtained suggested a progression of the human entorhinal cortex from the peripheral to the central subareas, with the intermediate subarea (281) as the most complete entorhinal subdivision. Compared with non-human primates, the human ERC not only retains the basic periallocortical organization but also demonstrates further evolution. Taken together with available experimental data on the connectivity of this brain region, these results provide an anatomical basis for evaluating the ERC in human behavior. PMID- 9408037 TI - A qualitative and quantitative analysis of the entorhinal cortex in schizophrenia. AB - The entorhinal cortex (ERC) has been implicated in schizophrenia by a number of studies. There is anatomical observation of neuronal heterotopias in the rostral ERC, which is consistent with a hypothesis of neurodevelopmental abnormalities in this disease. In view of the significant cytoarchitectonic variation of the ERC throughout its rostro-caudal extent, we performed a detailed subareal analysis of the rostral two-thirds of the entorhinal cortex (ERCr) in 14 postmortem schizophrenic brains and 14 matched controls (mean ages of 48 and 47 respectively). This systematic evaluation included both a qualitative microscopic analysis of morphogenetic anomalies that would be consistent with neurodevelopmental pathology and quantitative measurements of total neuronal number, average neuronal density, laminar volume and laminar depth from the cortical surface in cytoarchitectonically matched subareas of schizophrenic and control brains. Parcellation of the entire ERC on the basis of cytoarchitectonic criteria identified five distinct regions, similar to those described in the macaque, except that in the human brain three of the regions were further divisible into two or three subareas, yielding nine distinct cellular compartments. Five rostral areas, prorhinal (Pr), lateral (28L), intermediate rostral and caudal (281r and 281c), and sulcal (28S), comprise the ERCr. Gross and microscopic examination of these subdivisions throughout the ERCr failed to reveal laminar disorganization in any of the schizophrenic brains. The brains also did not differ significantly with respect to total neuronal number, total volume and neuronal density per laminar and subareal subdivision, or laminar thickness per entorhinal subarea. However, neuronal number and density were reduced by 12-18% in Pr and 28L, suggesting that mild quantitative abnormalities may exist in the ERCr and might possibly be revealed in a larger sample of schizophrenic brains. We have failed to confirm previous reports of laminar disorganization in the ERCr in brains of patients with schizophrenia; to the extent that this region is implicated in schizophrenia, the structural changes are likely to consist of more subtle cellular disturbances. PMID- 9408038 TI - Altered development of prefrontal neurons in rhesus monkeys with neonatal mesial temporo-limbic lesions: a proton magnetic resonance spectroscopic imaging study. AB - Focal brain damage occurring early in development can have widespread repercussions throughout the developing brain. In living adult rhesus monkeys, we studied the long-term effects of early mesial temporo-limbic (MTL) lesions on prefrontal cortex (PFC) neurons using proton magnetic resonance spectroscopic imaging (1H-MRSI), an in vivo neurochemical assay technique for measuring signals from metabolites such as N-acetyl-aspartate (NAA, a neuronal marker), choline containing compounds (CHO) and creatine + phosphocreatine (CRE). Six monkeys (NL) had undergone surgical ablation of MTL structures within 3 weeks of birth, six monkeys received the same lesion at approximately 5 years of age and six monkeys were normal controls. We found significant bilateral reductions of NAA relative signals exclusively in the PFC of the NL group in comparison with either of the other groups. Our results indicate that neonatal MTL damage specifically affects PFC neurons of adult monkeys as indicated by a reduction of NAA. The basis of this effect involves developmental processes as implicated by two arguments: analogous damage during adulthood does not have the same effect; NAA in the healthy brain increases during development. This finding may have implications for understanding developmental aspects of prefrontal-temporolimbic connectivity, and the reduction of NAA levels observed in prefrontal cortex of patients with schizophrenia. PMID- 9408039 TI - Maps of complex motion selectivity in the superior temporal cortex of the alert macaque monkey: a double-label 2-deoxyglucose study. AB - The superior temporal sulcus (STS) of the macaque monkey contains multiple visual areas. Many neurons within these regions respond selectively to motion direction and to more complex motion patterns, such as expansion, contraction and rotation. Single-unit recording and optical recording studies in MT/MST suggest that cells with similar tuning properties are clustered into columns extending through multiple cortical layers. In this study, we used a double-label 2-deoxyglucose technique in awake, behaving macaque monkeys to clarify this functional organization. This technique allowed us to label, in a single animal, two populations of neurons responding to two different visual stimuli. In one monkey we compared expansion with contraction; in a second monkey we compared expansion with clockwise rotation. Within the STS we found a patchy arrangement of cortical columns with alternating stimulus selectivity: columns of neurons preferring expansion versus contraction were more widely separated than those selective for expansion versus rotation. This mosaic of interdigitating columns on the floor and posterior bank of the STS included area MT and some neighboring regions of cortex, perhaps including area MST. PMID- 9408040 TI - Responses of macaque inferior temporal neurons to overlapping shapes. AB - We tested whether macaque inferior temporal neurons can signal the presence of their preferred shape independently of other shapes simultaneously present, by comparing the responses and selectivity of TE neurons to shapes (figures), either presented in isolation or overlapping another shape (background). The two overlapping shapes differed in color or texture and thus were easily segmentable. We found that the response and selectivity of TE neurons to the figure can be dramatically altered, most commonly reduced, when the figure is overlapping the background. This reduction in response was also present when the monkey was required to actively discriminate the figures from varying backgrounds during recording. The level of suppression depended on the shape of the background and on whether or not figure and background can be discriminated. These results indicate that responses of TE cells are not only determined by the properties of the figures but also are influenced by the properties of stimuli in the background. PMID- 9408041 TI - Modulation of connectivity in visual pathways by attention: cortical interactions evaluated with structural equation modelling and fMRI. AB - Electrophysiological and neuroimaging studies have shown that attention to visual motion can increase the responsiveness of the motion-selective cortical area V5 and the posterior parietal cortex (PP). Increased or decreased activation in a cortical area is often attributed to attentional modulation of the cortical projections to that area. This leads to the notion that attention is associated with changes in connectivity. We have addressed attentional modulation of effective connectivity using functional magnetic resonance imaging (fMRI). Three subjects were scanned under identical stimulus conditions (visual motion) while varying only the attentional component of the task. Haemodynamic responses defined an occipito-parieto-frontal network, including the, primary visual cortex (V1), V5 and PR A structural equation model of the interactions among these dorsal visual pathway areas revealed increased connectivity between V5 and PP related to attention. On the basis of our analysis and the neuroanatomical pattern of projections from the prefrontal cortex to PP we attributed the source of modulatory influences, on the posterior visual pathway, to the prefrontal cortex (PFC). To test this hypothesis we included the PFC in our model as a 'modulator' of the pathway between V5 and PP, using interaction terms in the structural equation model. This analysis revealed a significant modulatory effect of prefrontal regions on V5 afferents to posterior parietal cortex. PMID- 9408042 TI - Hypothermia-related deaths -- Virginia, November 1996-April 1997. AB - Hypothermia is defined as a central or core body temperature of < or =95 F (< or =35 C) and is a medical emergency. Persons with hypothermia are at high risk for death. Although hypothermia-related deaths are common during winter months in states characterized by cold winters (e.g., Alaska and North Dakota) and with mountainous or desert terrain (e.g., Arizona and New Mexico), hypothermia and associated deaths also occur in states with milder climates. For example, during November 1996-April 1997, the Chief Medical Examiner's Office in Virginia identified 20 deaths caused by hypothermia; of these, 11 (55%) were among men and decedents ranged in age from 22 to 86 years (mean: 63 years). This report describes selected cases of hypothermia-related deaths in Virginia during November 1996-April 1997 and summarizes hypothermia-related deaths in the United States during 1979-1994. PMID- 9408043 TI - Measles outbreak -- Romania, 1997. AB - During December 1, 1996-September 30, 1997, a total of 20,034 cases of measles (incidence: 88.7 per 100,000 population) were reported to the Ministry of Health (MOH) in Romania (Figure 1); 13 cases were fatal. The outbreak began in December 1996, peaked in May 1997, then declined. Cases occurred in the capital (Bucharest) and all 40 other districts (1996 total population: 22.6 million). District-specific attack rates were highest in the northwest and lower in the south, ranging from 10 to 258 cases per 100,000 population. This report describes the investigation of this epidemic by MOH and estimates the efficacy of measles vaccine using the screening method; the findings of the investigation suggest that high routine vaccination coverage with a single dose of measles vaccine with an estimated efficacy of 77%-90% was not sufficient to prevent periodic outbreaks of measles. PMID- 9408044 TI - Update: respiratory syncytial virus activity -- United States, 1997-98 season. AB - Respiratory syncytial virus (RSV), a common cause of winter outbreaks of acute respiratory disease, results in an estimated 90,000 hospitalizations and 4500 deaths each year from lower respiratory tract disease among infants and young children in the United States. Outbreaks occur annually throughout the country. RSV activity in the United States is monitored by the National Respiratory and Enteric Virus Surveillance System (NREVSS), a voluntary, laboratory-based system. This report summarizes trends in RSV reported by NREVSS for July 1992-June 1997 and presents provisional surveillance results for July-November 1997. These data indicate onset of widespread RSV activity for the 1997-98 season. PMID- 9408045 TI - Vaccination levels among Hispanics and non-Hispanic whites aged > or =65 years -- Los Angeles County, California, 1996. AB - An estimated 90% of deaths from pneumonia and influenza occur each year in the United States among adults aged > or = 65 years. Despite the substantial impact of these and other vaccine-preventable diseases on older adults, national vaccination levels are suboptimal and disproportionately lower among some racial/ethnic minorities than among others. For example, in 1995, influenza and pneumococcal vaccination rates for older Hispanics (50.0% and 24.2%, respectively) were substantially lower than those for non-Hispanic whites (60.1% and 37.4%, respectively). To develop and implement community-based activities to increase vaccination levels among older Hispanic adults in Los Angeles County, California, the Edward R. Roybal Institute for Applied Gerontology at California State University, Los Angeles, formed a community consortium involving multiple public and private organizations. During August-November 1996, this consortium, in collaboration with the Center for the Study of Latino Health at the University of California, Los Angeles (UCLA), conducted a telephone survey to assess vaccination knowledge, attitudes, and practices of older Hispanic adults and to provide baseline information for developing interventions. This report summarizes the results of the initial assessment conducted in two geographic areas; the findings document low vaccination levels among the populations surveyed and race/ethnicity-specific differences in barriers to vaccination and places where vaccinations were received. PMID- 9408046 TI - Human monkeypox -- Kasai Oriental, Democratic Republic of Congo, February 1996 October 1997. AB - Human monkeypox is a severe smallpox-like illness caused by monkeypox virus (MPV); monkeypox occurs in sporadic outbreaks, and infection is enzootic among squirrels and monkeys in the rainforests of western and central Africa. In 1996, cases of monkeypox were reported from villages in the Katako-Kombe Health Zone, Kasai Oriental, Zaire (i.e., Democratic Republic of Congo). The World Health Organization (WHO), in collaboration with CDC, investigated this outbreak and identified 92 suspected cases with onset during February 1996-February 1997, and isolated MPV from lesions of active cases. Cases continued to be reported, and a new investigation was initiated by WHO and CDC in October 1997. This report summarizes the results of the field investigation, which indicate that this is the largest human monkeypox outbreak ever recorded. PMID- 9408047 TI - Temporal trends in the incidence of birth defects -- United States. AB - Through CDC's Birth Defects Monitoring Program (BDMP), a total of 161 categories of birth defects are analyzed quarterly to determine increases or other unusual trends. Sixteen of these malformations have been selected for review in this report because they occur in sufficient numbers to provide relatively stable rates, the coding categories for them are relatively homogeneous, and they represent defects of different organ systems. PMID- 9408048 TI - Newborn screening for cystic fibrosis: a paradigm for public health genetics policy development. Proceedings of a 1997 workshop. AB - Cystic fibrosis (CF) is a genetic disease that can be detected in newborn infants (i.e., those aged < or = 1 month) by immunotrypsinogen testing. The sensitivity and specificity of such testing can now be improved as a result of the recent discovery of the Cystic Fibrosis Transmembrane Conductance Regulatory (CFTR) gene. Although limited CF screening for newborns has been used since the 1980s, the clinical, social, and economic outcomes of population-based screening are controversial. During January 1997, a workshop was convened at CDC in Atlanta, Georgia to discuss the benefits and risks associated with screening newborns for CF and to develop public health policy concerning such screening. The workshop planning committee comprised representatives from CDC, the Cystic Fibrosis Foundation, the National Institutes of Health, and the University of Wisconsin. Experts in the fields of CF, public health, the screening of newborns, and economics also contributed to discussions. Workshop participants addressed a) benefits and risks, b) laboratory testing, and c) economics concerning the implementation of routine CF screening for newborns. Summaries of these discussions and the resulting workshop recommendations are presented in this report. These recommendations, developed by workshop participants, will be useful to medical and public health professionals and state policymakers who are evaluating the merits of population-based screening of newborns for CF. PMID- 9408049 TI - The somatostatin receptor-adenylate cyclase system in rat pancreatic acinar membranes after temporary pancreaticobiliary duct ligation. AB - The mechanism whereby somatostatin (SS) produces beneficial effects in established pancreatitis induced by pancreaticobiliary duct ligation (PBDL) is still not clear. The aim of the work was to evaluate the possibility of a direct action of SS on pancreatic acinar cells from rats with acute pancreatitis. For this purpose, we studied the SS-receptor-adenylate cyclase system in pancreatic acinar membranes from both, control rats and rats with experimentally induced acute pancreatitis. On the other hand, it has been reported that cholecystokinin (CCK) diminishes the number of SS receptors in pancreatic acinar cells. Proglumide, a CCK receptor antagonist reduces the severity of acute pancreatitis in the rat. Therefore, we have also examined the effect of proglumide on the somatostatinergic system in controls and rats with acute pancreatitis. Fourteen hours after PBDL, the SS receptors, the capacity of the SS analogue SMS 201-995 to inhibit forskolin-stimulated adenylate cyclase activity and PTX-catalyzed [32P] ADP-ribosylation of the alpha1 subunits of Gi proteins could not be detected in pancreatic acinar membranes. One month after reopening the closed pancreaticobiliary duct (PBD), the pancreas showed regeneration of acinar cells, and the above-mentioned parameters were significantly lower than in the control group. Two months after reopening the closed PBD, all these parameters had returned to control values. The administration of proglumide (20 mg/kg i.p.), a cholecystokinin receptor antagonist, accelerated pancreatic regeneration and approached all these parameters to control values one month after reopening the closed PBD. The present study suggests that the beneficial effects of SS on established pancreatitis induced by PBDL may not be due to a direct action of the peptide on pancreatic acinar cells at least at 14 hours after PBDL. In addition, these findings suggest that in established pancreatitis the effect of proglumide on the SS receptor-adenylate cyclase system could be due to its action on pancreatic regeneration. PMID- 9408050 TI - Kappa opiate agonist RU 51599 inhibits vasopressin gene expression and osmotically-induced vasopressin secretion in vitro. AB - Kappa (kappa) opioid agonists induce a water diuresis and inhibit vasopressin (AVP) secretion. Hypothalamic and neurohypophysial sites have both been implicated in the response. The present study was designed to ascertain if kappa agonist inhibition of osmotically-stimulated AVP secretion is associated with parallel changes in AVP gene expression. Experiments were performed using the selective kappa-agonist RU 51599 (RU) in compartmentalized hypothalamo neurohypophysial explants. When added to either the hypothalamus or the neural lobe, RU dose dependently inhibited osmotically-induced AVP secretion that was reversed by the highly selective kappa-antagonist nor-binaltorphimine (nor-BNI) only at the hypothalamic, not the neurohypophysial level. AVP mRNA content paralleled the changes in AVP secretory rate induced by hypothalamic kappa agonism. AVP mRNA levels were unaltered when RU was applied to the neural lobe. Neurohypophysial AVP content did not change. These data indicate that hypothalamic kappa-agonism inhibits osmotically induced AVP secretion and that a non-kappa1 opiate receptor mediates posterior pituitary opioid inhibition of AVP release. Neural or receptor inputs to the hypothalamus or magnocellular cell body may downwardly modulate AVP mRNA content by altering AVP gene transcription and/or message stability. Inhibition of AVP release directly at the neurohypophysis can be uncoupled from the cellular mechanisms that generate changes in AVP mRNA content. PMID- 9408051 TI - In vivo interaction of baclofen, TRH and serotonin on PRL and TSH secretion in the developing and adult male and female rats. AB - Gamma-aminobutyric acid (GABA) is involved in the neural control of hypophyseal hormones, including PRL and TSH. In the present work we investigated the ontogeny of the effect of baclofen, a GABA B agonist, on basal PRL and TSH release and in the presence of releasing stimulus which act at two different levels: TRH, at the hypophyseal level, and serotonin, at the central nervous system. Ages studied were 4, 12, 20, 28-29, 37-38 day-old and adult male and female animals. Rats of each age and sex were separated in groups and each group received two intraperitoneally injections, one 45 minutes after the other: saline-saline, saline-TRH, baclofen-saline, baclofen-TRH, saline-serotonin or baclofen serotonin. Rats were decapitated 15 minutes after the last injection and serum hormones were measured by RIA. Baclofen (7 mg/kg) significantly elevated basal prolactin levels at 4, 12 and 20 days of age and the stimulating effect increased with age. At 28 days of age baclofen significantly inhibited PRL whereas from 38 days of age onwards it had no effect on basal PRL levels. No sex differences were evident. Interaction of TRH (4 microg/kg) and baclofen on PRL secretion resulted in an additive effect on days 4 and 12, this effect was not observed when baclofen was administered with serotonin (10 mg/kg). In 28 day-old and older animals baclofen completely blunted the PRL releasing effect of TRH or serotonin. Again, no sex differences were observed. With regard to TSH, baclofen did not alter either basal or TRH stimulated TSH secretion regardless of sex and age. The present experiments indicate that GABA B receptors are involved in the regulation of basal and stimulated PRL secretion from the first days of life to adulthood. Receptor activation results in stimulation or inhibition of PRL release depending on the age of the animals and the site of action. This GABA B regulation of PRL secretion is sex independent. In contrast, pituitary GABA B receptors do not seem to be involved in the regulation of TSH secretion. PMID- 9408052 TI - The antigenic sites of trichosanthin, a ribosome-inactivating protein with multiple pharmacological properties. AB - Trichosanthin (TCS) fragments were produced by Tn1000 deletion mutagenesis and by cyanogen bromide (CNBr) cleavage and their immunoreactivity was examined by incubating with various antibodies. Twelve C-terminally truncated TCS variants were successfully synthesized under the control of a T7 RNA driven promoter. The smallest antigenic fragment mapped corresponded to the N-terminal 20 amino acids (aa). Six CNBr fragments of TCS were created and identified by electrospray mass spectrometry and N-terminal sequencing. Three antigenic fragments corresponding to aa 1-72, 101-152 and 153-246, respectively were mapped. Fragments corresponding to aa 1-72 and 153-246 were immunoreactive to the same monoclonal antibody showing they are components of a discontinuous epitope. On the other hand, the fragment containing aa 73-100 was not detected by any of the antibodies used. PMID- 9408053 TI - Monoclonal antibodies as surrogate receptors in a high throughput screen for compounds that enhance insulin sensitivity. AB - Monoclonal antibodies (MoAbs) were made to a known insulin sensitivity enhancer (ISE) compound, CS-045. The MoAbs were characterized with respect to binding other known thiazolidinedione ISE compounds using a CS-045 labeled with b phycoerythrin in a competitive particle concentration fluorescence immunoassay (PCFIA). By comparing the rank order of IC50 values for each compound to its respective potency as an ISE, one MoAb (13E3) was selected for further characterization. This MoAb was also used as a surrogate receptor in a high throughput screen to identify novel compounds that compete for binding to CS-045. Some of the hits were found to have efficacy in reducing blood glucose. Subsequently, another group reported that several compounds with the core thiazolidinedione structure of the ISE compounds bound with high affinity to peroxisome proliferator-activating receptors (PPAR). Therefore, we used the MoAb assay to test these and other compounds that are known to bind to PPARgamma and noted crossreactivity with some of the compounds. PMID- 9408054 TI - Heterologous expression of carbonyl reductase: demonstration of prostaglandin 9 ketoreductase activity and paraquat resistance. AB - Transfection of murine NIH3T3 fibroblasts with a pSV2-derived eukaryotic expression vector for human cytosolic carbonyl reductase (E.C. 1.1.1.141) resulted in clones with increased carbonyl reductase activity as demonstrated by an elevation in cellular NADPH-dependent alcohol (menadione) reductase activity. Prostaglandin 9-ketoreductase (9KR) activity, previously noted only in purified enzyme preparations, was also elevated. Although the cellular molar capacity of 9KR activity was less than menadione reductase activity (picomoles versus nanomoles per mg of protein), when compared to endogenous activity there was a greater relative increase in 9KR activity as compared to menadione activity (10 fold increase versus 3 fold). Thus, the 9KR properties of carbonyl reductase may have a physiologic role in prostaglandin regulation. Most transgenic clones lost their enhanced carbonyl reductase activity despite continuous selection, but two clones retained enhanced enzyme activity. RNA analysis indicated that these two murine clones expressed human carbonyl reductase mRNA. These two clones overexpressing carbonyl reductase did not display resistance to menadione, in agreement with a previous report. There was, however, a demonstrable increase in resistance to paraquat of a magnitude similar to that previously noted with transgenic cell lines overexpressing manganese superoxide dismutase. PMID- 9408055 TI - Effect of a carboxy-terminal fragment of the Alzheimer's amyloid precursor protein on expression of proinflammatory cytokines in rat glial cells. AB - To explore factors involved in the induction of cytokines that may contribute to the pathogenesis of Alzheimer's disease (AD), the effect of a carboxy terminal 105 amino acid fragment (CT105) of the amyloid precursor protein (APP) on the gene expression of proinflammatory cytokines, IL-1beta and IL-6 was determined in cultured rat cortical glial cells in comparision to amyloid beta protein (A beta). Cells were incubated with 1 microM of insoluble CT105 aggregates or aged A beta1-40 peptide deposits which were mainly composed of stable monomeric and dimeric forms as assessed on Western blots. The levels of mRNAs were analyzed by reverse transcription polymerase chain reaction (RT-PCR). Highest levels of both IL-1beta and IL-6 transcripts were detected in the culture exposed to CT105 aggregates for 4 days. CT105 aggregates markedly increased IL-1beta mRNA level by 3.5 fold of the control level and this effect was more potent than that produced by aged A beta1-40 peptides. Furthermore, CT105 strongly induced accumulation of IL-6 mRNA level by 2 fold of the value potentiated by A beta1-40. Such induction was not observed with A beta 12-28 treatment. On the other hand, CT105 did not significantly alter either APP or glial fibrillary acidic protein (GFAP) mRNA levels. These results together imply that CT peptide besides its cytotoxic potency may act as a potent immunological component, strongly inducing both IL 1beta and IL-6 mRNA levels in the cultured glial cells. This CT peptide associated exacerbation of cytokine expression may be in part responsible for chronic inflammation linked to slowly progressive neurodegeneration characteristic to AD. PMID- 9408057 TI - Ethanol-induced apoptosis in human HL-60 cells. AB - The mechanisms responsible for aberrant immune function associated with chronic ethanol use remain obscure, but a decrease in monocyte numbers is often reported for individuals who chronically abuse ethanol. We investigated, using human HL-60 promyelocytic cell line, the possibility that ethanol induces apoptosis which contributes to decreased monocyte numbers. Characteristic features of apoptosis were observed 4 days after ethanol treatment, as documented by increased DNA fragmentation; enhanced expression of phosphatidylserine, an early marker of apoptosis; and the appearance of a hypodiploid apoptotic cell population identified by flow cytometry analysis of the cell cycle. Treatment with the protein kinase C inhibitor, GF 109203X, potentiated ethanol-induced apoptosis. Direct induction of human HL-60 cell apoptosis by ethanol and potentiation of ethanol-induced apoptosis by inhibiting protein kinase C provides a partial explanation for the cytotoxic effects of ethanol on hematopoietic progenitor cells and establishes a link between inhibiting protein kinase C activity and ethanol-induced apoptosis. PMID- 9408056 TI - A tumor cell growth inhibitor from Polygonum hypoleucum Ohwi. AB - Polygonum hypoleucum Ohwi (P. hypoleucum Ohwi) has been used as a Chinese medicine for a long time. In the present study, four anthraquinones, emodin, emodin 1-O-beta-D-glucoside (49A), physcion (62A), and physcion 1-O-beta-D glucoside (50A) were identified from P. hypoleucum Ohwi and their inhibitory effects on various tumor cells proliferation were investigated. On a percentage basis, emodin had the highest suppressing activity on the various tumor cells proliferation. At 10 microg/ml, the percentage inhibition on K562 cells proliferation for emodin, 49A, 62A, and 50A were 97+/-3.4%, 18+7.3%, 24+/-3.6%, and 31+/-8.9%, respectively. However, inhibitory activities of 10 microg/ml of emodin, 49A, 62A, or 50A on Raji cells proliferation were 98+/-5.0%, 25+/-5.0%, 22+/-3.2%, and 28+/-4.3%, respectively. It was also found that the both C1 and C3 positions of emodin were important for antitumor action. The IC50s of emodin, 49A, 62A, and 50A on various tumor cells were also calculated. The IC50 of emodin on K562 cells was significantly lower than on Raji, HeLa, Calu-1, Wish, and Vero cells (1.5+/-0.2 vs. 2.8+/-0.4 microg/ml, P < 0.01 ;1.5+/-0.2 vs. 8.4+/-1.6 microg/ml; 1.5+/-0.2 vs. 8.9+/-1.0 microg/ml; 1.5+/-0.2 vs. 8.7+/-0.5 microg/ml; 1.5/-0.2 vs. 3.5+/-0.12 microg/ml; P < 0.001). The results indicated that K562 and Raji cells were more sensitive to emodin treatment. Cell viability test indicated that inhibitory effect of emodin on various tumor cell lines was not through direct cytotoxicity. It suggested P. hypoleucum Ohwi included a tumor cell growth inhibitor. PMID- 9408058 TI - Comparative study of toborinone (OPC-18790) and milrinone on energy metabolism in microembolized guinea pig hearts. AB - The effects of toborinone (OPC-18790) and milrinone on cardiac function and energetics were compared in microembolized guinea pig hearts. Male guinea pig hearts were perfused according to the Langendorff method and microembolization was induced by injecting microspheres. The hearts were then treated with toborinone (10 microM), milrinone (4 microM), and vehicle. Energy metabolism in hearts was assessed by 31-phosphorus magnetic resonance spectroscopy (31P-MRS). Microembolization produced a decrease in coronary perfusion flow (CPF), left ventricular developed pressure (LVP), and peak LVdP/dt by about 50% concomitantly with a decrease in creatine phosphate (PCr) and ATP and an increase in inorganic phosphate (Pi) and Pi/PCr ratio. Toborinone and milrinone increased peak LVdP/dt, an index of contractility, by 15 +/- 2% and 18 +/- 3%, respectively. Milrinone increased heart rate (HR) by 22 +/- 4% but toborinone did not change HR. Toborinone did not change PCr, ATP, Pi, Pi/PCr, and intracellular pH (pHi) compared with the vehicle. On the other hand, milrinone decreased PCr and increased Pi and Pi/PCr compared with toborinone or vehicle. These results suggest that the different effects between toborinone and milrinone on energy metabolism in microembolized hearts may be due to the difference of chronotropic action between these drugs. Thus toborinone, a positive inotropic agent without chronotropic action, may be effective in acute treatment of ischemic heart failure. PMID- 9408059 TI - Increased apoptosis of adult rat lymphocytes after single neonatal vitamin A treatment (hormonal imprinting). A flow cytometric analysis. AB - Newborn rats were treated with a single dose of vitamin A (retinol) and apoptosis of peripheral lymphocytes was studied by flow cytometry in adult age. Vitamin A treatment (hormonal imprinting) caused a moderate, however significant elevation in the number of apoptotic lymphocytes after three months. Dexamethasone or Concanavalin-A alone did not influence apoptosis significantly. However, in the neonatally retinol treated rats dexamathasone significantly elevated the quantity of apoptotic lymphocytes related to the control or Concanavalin-A treated control cells. The results call attention to the prolonged effect of hormonal imprinting in a new index and to the possible dangerous effects in human, neonatally treated with vitamin A. PMID- 9408060 TI - Glutamate metabotropic receptors modulate the expression of in vitro epileptiform activity in rat hippocampal slices. AB - The effects of the mixed class I and II mGLUR agonist (+) 1S,3R-trans-amino cyclopentane-1,3-dicarboxylic acid (ACPD) and antagonists (+) alpha-methyl-4 carboxyphenylglycine (MCPG) and L-2-amino-3-phosphonopropionic acid (L-AP3) on the basal neuronal excitability and on the expression of in vitro epileptiform activity produced by the convulsant drugs picrotoxin and penicillin were investigated in rat hippocampal slices. The duration of the CA1 epileptiform bursting produced by 0.05 mM picrotoxin or 1 mM penicillin or 0.075 mM ACPD was significantly (p<0.05) and dose-dependently decreased by 0.3-0.5 mM MCPG or L AP3, but not by 0.05 mM ACPD. The data demonstrate an involvement of class I and II mGLURs in the basal neuronal excitability and in the expression of in vitro epileptiform activity produced by some convulsants. PMID- 9408061 TI - IgA nephropathy: 30 years on. AB - Thirty years since its discovery, IgA nephropathy (IgAN) has turned out to be the commonest primary glomerulonephritis around the world. The major presenting symptom is haematuria, with or without proteinuria. IgAN was initially considered a benign disease, but long-term follow-up studies have shown the course to be slowly progressive with up to 50% of the patients developing terminal renal failure after several decades. The aetiology and pathogenesis are still unclear, but IgAN seems to be associated with a dysregulation of the immune response which is not very well understood. There is today no specific treatment. Effective blood pressure control is the cornerstone of therapy. PMID- 9408062 TI - The clinical course and outcome in patients with acute ischaemic stroke and transient ischaemic attack in relation to severe carotid disease. AB - OBJECTIVES: To determine whether a severe ipsilateral carotid lesion influences the patient's clinical course and outcome in the acute phase of an ischaemic stroke or transient ischaemic attack (TIA). DESIGN: An evaluation of prospectively-collected and recorded clinical information and sonographic findings in patients treated in a stroke unit. SUBJECTS: Two hundred and sixty six consecutive patients with acute ischaemic stroke or TIA in the carotid territory. MAIN OUTCOME VARIABLES: A sonography of the carotid arteries was performed in all cases. Stroke symptoms were continuously monitored and progression and recurrence particularly looked for. Patient outcome at discharge was also recorded. RESULTS: An ipsilateral carotid stenosis of > or = 70% was found in 11%, and 4% had an occlusion. These patients had no statistically significant increase in the rate of progression or recurrence of stroke during the acute phase. Furthermore, they did not become more disabled or in need of extended institutional care. CONCLUSIONS: Severe ipsilateral carotid disease does not significantly influence the acute clinical course or outcome in ischaemic stroke patients and there seems to be no need for urgent sonography and treatment. PMID- 9408063 TI - Proinflammatory cytokines, measured in a mixed population on arrival in the emergency department, are related to mortality and severity of disease. AB - OBJECTIVES: To investigate whether serum levels of tumour necrosis factor alpha (TNF-alpha) and interleukin 6 (IL-6) are related to mortality and severity of disease in patients admitted to the Emergency Department (ED). DESIGN: Prospective cohort study. SETTING: Emergency Department of a tertiary university hospital. SUBJECTS: A total of 140 patients admitted to the ED, representing common acute medical diseases, such as stroke, obstructive lung disease, heart failure, myocardial infarction, angina pectoris, infectious diseases and acute abdominal disorders. MAIN OUTCOME MEASURES: APACHE II score at admission, hospital mortality and length of stay in hospital (LOS). RESULTS: A moderate rise in cytokine levels (IL-6; 50-300 ng L(-1), TNF-alpha; 10-70 ng L[-1]) was found in all diagnosis-groups, with the most pronounced elevation seen in patients with acute abdominal disorders (up to 6900 ng L[-1]). IL-6 on arrival to the ED was significantly correlated to the APACHE II score (r = 0.48, P < 0.0001), LOS (r = 0.36, P < 0.0001) and was elevated in nonsurvivors (n = 9) compared to those who did survive. TNF-alpha on arrival showed a significant correlation to LOS (r = 0.36, P < 0.0001) and APACHE II (r = 0.41, P < 0.0001), but was not associated to later mortality. CONCLUSIONS: Serum levels of proinflammatory cytokines collected at admission to the ED were related to the severity of disease and hospital mortality. PMID- 9408064 TI - The ineffectiveness of cyclical oral clodronate on bone mineral density in glucocorticoid-treated patients with giant-cell arteritis. AB - OBJECTIVES AND DESIGN: The aim of the present study was to determine whether cyclic oral administration of clodronate, a bisphosphonate, every second month prevents rapid bone loss during the first year of glucocorticoid treatment in patients with giant-cell arteritis (GCA). The trial was designed as a prospective double blind study, assessing total body mineral content (BMC) and bone mineral density (BMD) using DXA technique. Supplementation of calcium was given to both groups of patients. SETTING: The outpatient clinics of the rheumatic and infectious diseases of Sahlgren University Hospital of the city of Goteborg on the west coast of Sweden. SUBJECTS: Twenty-seven patients with confirmed GCA were consecutively included during a 15-month period. RESULTS: An early influence on bone turnover was found with a temporary decrease in BMC after six months of glucocorticoid treatment, which was normalized after 12 months in both study groups. No significant differences between the patients given clodronate and calcium and the controls, who got supplementation with calcium alone, was observed at any assessment point. However, there was a significant and prolonged depression of the osteocalcin levels in the clodronate-treated patients. CONCLUSIONS: Oral administration of clodronate in a moderately high dose given cyclically every other month had no additive effect on BMD compared with calcium supplementation alone during the first year of glucocorticoid treatment. A larger material might have revealed some differences between the categories. In most patients with GCA, however, the BMD seems to recover after one year of glucocorticoid treatment, provided there is good control of the inflammation and patients are kept physically active. It needs to be elucidated whether there are subsets of patients who might benefit from bone sparing agents: women near menopause with a high turnover rate of bone, individuals who have low BMD from the start of glucocorticoid treatment or patients requiring high doses of glucocorticoids during a long period of time. PMID- 9408065 TI - The effects of thromboxane A2 inhibition (picotamide) and angiotensin II receptor blockade (losartan) in primary Raynaud's phenomenon. AB - OBJECTIVES: To assess the role of thromboxane A2 and of angiotensin II in patients with primary Raynaud's phenomenon. DESIGN: After an eight-day run-in period, the patients were enrolled in a single-blind, cross-over, study. SETTING: Patients were examined at the Ambulatory for Microcirculatory Diseases of the Clinic of Internal Medicine, University Hospital, Verona. SUBJECTS: Fifteen subjects affected by primary Raynaud's phenomenon were included. MAIN OUTCOME MEASURES: A piezoelectric plethysmography to evaluate the distensibility of the digital arteries as the ratio between peak time (PT) and total time (TT), and an oscillometric blood pressure recorder were used after the run-in period, and after a two-week course of picotamide (300 mg b.i.d., i.e. two times daily) or losartan (12.5 mg once daily) with an interval of a week of placebo between the active treatments. The tests were performed after every treatment in basal condition and during mental stress. The patients reported in a diary the number and the severity (from 0 to 4 +) of the vasospastic crises. RESULTS: The change in TP/TT ratio appeared statistically significant only after losartan treatment, both in basal condition and during mathematical stress. Both pharmacological treatments, with respect to placebo, showed an improvement of the scores, derived from the number and severity of vasospastic attacks, but only the therapy with losartan determined a statistically significant improvement. CONCLUSIONS: The inhibition of the type 1 receptor for angiotensin II seems highly effective in the reduction of the vasospastic crises in the subjects with primary Raynaud's phenomenon. According to our experience, losartan could be used more extensively in the treatment of these patients besides arterial hypertension. PMID- 9408066 TI - Low circulating leptin levels in protein-energy malnourished chronically ill elderly patients. AB - OBJECTIVE: To evaluate serum leptin, a fat cell-derived protein, levels in relation to the malnutrition often observed in chronic disease. DESIGN: A comparison of circulating leptin concentrations in malnourished chronically ill elderly and in age-matched controls. SETTING: A university-affiliated teaching hospital in Stockholm, Sweden. SUBJECTS: Nineteen protein-energy malnourished elderly patients (74 +/- 1 years) with various chronic nonmalignant diseases and 18 healthy controls (72 +/- 1 years). MAIN OUTCOME MEASURES: Serum leptin levels measured by radioimmunoassay technique, nutritional status as expressed by body mass index (kg m[-2]), triceps skin fold, arm muscle circumference and serum albumin, and serum orosomucoid concentrations indicating inflammatory status. RESULTS: Patients and controls displayed body mass indexes of 17.4 +/- 0.7 and 25.0 +/- 1.1 (P < 0.001), respectively. Triceps skin fold (TSF) measurements revealed a pronounced fat depletion in the patients, being 8.5 +/- 0.9 and 22.3 +/- 1.5 mm (P < 0.001) in female and 6.1 +/- 0.7 and 10.8 +/- 0.8 mm (P < 0.001) in male patients and controls, respectively. Patient serum leptin concentrations were less than half of the corresponding concentrations in the controls, 4.3 +/- 1.1 and 9.3 +/- 1.3 ng mL(-1)(P < 0.01), respectively. The highest leptin concentrations were registered in female controls, 12.1 +/- 1.6 ng mL(-1). The serum leptin levels in the controls correlated with TSF (r = 0.74: P < 0.001). No such correlation was found in the patients. CONCLUSIONS: Serum leptin levels were low and did not seem to be directly associated with fat and muscle depletion in elderly patients with chronic illness, whereas they appeared to be positively correlated to body fat in healthy elderly. PMID- 9408067 TI - The anti-oestrogen tamoxifen produces haemodilution in normal postmenopausal women. AB - OBJECTIVES: To report the effects of the anti-oestrogen tamoxifen on biochemical and haematological parameters. DESIGN: Randomized, double-blind comparison of tamoxifen 20 mg per day and placebo, over two years. SETTING: A university hospital. SUBJECTS: Forty-six healthy late-postmenopausal women (mean, SD time since menopause; 11, seven years). MAIN OUTCOME MEASURES: Blood specimens were drawn in the fasting state at baseline, six months and two years for measurement of haemoglobin, haematocrit, erythrocyte mean cell volume, mean erythrocyte haemoglobin, leucocyte count, platelet count, urea, electrolytes, creatinine and albumin. RESULTS: There was a significant decline in the haemoglobin concentration in the tamoxifen group (-4.4, 1.2 g/L; mean, SE) and its levels were lower in this group than in those receiving placebo (P = 0.004). Similarly, haematocrit, erythrocyte count and total leucocyte count were lower in those on placebo (P = 0.002, P = 0.01 and P = 0.01, respectively) and platelet count showed a similar trend (P = 0.08). In the tamoxifen group, the level of serum albumin fell significantly (-2.2, 0.4 g/L) and was lower throughout the study than that in the placebo group (P = 0.006). That of serum urea tended to fall ( 0.4, 0.2 mmol L) but the between-groups comparison was not significant (P = 0.18). CONCLUSIONS: These data suggest that tamoxifen exerts a haemodilutory effect in normal postmenopausal women. Since a similar effect has been reported in response to postmenopausal oestrogen therapy, it is likely that the observed changes represent another oestrogenic effect of tamoxifen in postmenopausal women. Haemodilution may contribute to the reduced incidence of cardiovascular disease reported in tamoxifen-treated women, and, therefore, its assessment in the new oestrogen agonists/antagonists being developed for cardiovascular disease prevention may be important. PMID- 9408068 TI - Does metformin increase the serum total homocysteine level in non-insulin dependent diabetes mellitus? AB - OBJECTIVE: The aim of this study was to estimate the effect of metformin on the serum total homocysteine level in non-insulin-dependent diabetes mellitus (NIDDM) patients. An elevated serum total homocysteine level is a risk factor for atherosclerosis. Metformin decreases serum vitamin B12, and may thereby indirectly increase the serum total homocysteine level. DESIGN: A cross-sectional study in a primary care setting. SUBJECTS, MAIN OUTCOME MEASURES: Fasting serum total homocysteine level was measured in 40 NIDDM patients who had received treatment with metformin (500-2550 mg per day) for at least six months, and in 71 NIDDM patients not treated with metformin and matched for sex, age (+/- 5 years), serum creatinine (+/- 5 micromol L[-1]) and current smoking habits. 'Exposed' patients were matched with 'nonexposed' patients. A two-way analysis of variance was performed. RESULTS: The mean serum total homocysteine level was 11.5 micromol L(-1) in the metformin-exposed patients and 10.6 micromol L(-1) in the nonexposed patients. Thus, the metformin-exposed patients had slightly higher serum total homocysteine levels (difference 0.8 micromol L(-1), 95% confidence interval (-0.4 2.0 micromol L[-1]). Results were similar in men and women. Finally, no dose response relationship between cumulative exposure to metformin (dose x duration of treatment) and the serum total homocysteine level could be demonstrated. CONCLUSION: We conclude that the effect of metformin on serum total homocysteine level in NIDDM patients, if any, is likely to be small. PMID- 9408069 TI - Improving empirical antibiotic treatment: prospective, nonintervention testing of a decision support system. AB - OBJECTIVES: Develop a problem-orientated and data-based decision support system (DSS) to improve empirical antibiotic treatment, and compare the performance of the system to that of the physician. DESIGN: The DSS was tested in a prospective, noninterventional, comparative cohort study. SETTING: University hospital in Israel. SUBJECTS: Consecutive patients (n = 496) in four departments of internal medicine suspected of harboring a moderate to severe bacterial infection. INTERVENTIONS: None. MAIN OUTCOME MEASURES: The percentage of appropriate empirical antibiotic treatments. RESULTS: Out of 496 patients included in the study, 219 had positive cultures or serological tests. The physicians prescribed inappropriate empirical antibiotic treatment in 91 of 219 patients (42%); whilst the recommendations of the system were inappropriate in 50 patients (23%) (P < 0.05). Superfluous treatment was prescribed in 15% of patients by the physician, and in 11% by the system. Out of the 91 patients given inappropriate treatment by the physician, the DSS advised treatment to which the pathogens were susceptible in 61 patients. The advantage of the DSS over the physician was most evident in multiresistant gram-negative isolates, enterococci and Staphylococcus aureus. Out of the 277 patients with negative cultures, the DSS advised narrower-spectrum antibiotic treatment than prescribed by the physicians in 27% of patients, and broader-spectrum in 13%. CONCLUSION: A problem-orientated, data-based DSS outperformed physicians in the choice of appropriate empirical antibiotic treatment, and recommended less broad-spectrum antibiotics. PMID- 9408070 TI - Narrow hips and broad waist circumferences independently contribute to increased risk of non-insulin-dependent diabetes mellitus. AB - OBJECTIVES: Patients with non-insulin-dependent diabetes mellitus (NIDDM) have been shown to be more obese and have higher waist-to-hips circumference ratios compared to nondiabetics. In this study, we tried to dissociate obesity, waist and hip circumference from NIDDM. DESIGN: A cross-sectional population-based case control study. SETTING: General population. SUBJECTS: CONTROLS: 5887 men and 7018 women, aged 20-59 years, without known diabetes or hyperglycaemia. CASES: 93 men and 66 women were diagnosed with NIDDM. MAIN OUTCOME MEASURES: We predicted waist and hips' circumference from the body mass index, weight/height2, (BMI) on the basis of linear regression. Differences between observed and expected values (residuals) of waist and hip circumference were categorized into tertiles. The relative odds of having NIDDM in tertiles of waist and hip residuals (middle tertile as reference) were calculated by multiple logistic regression analysis adjusted for each other and for age, smoking, physical activity, alcohol consumption and education. RESULTS: NIDDM was most prevalent in men and women who had larger waists and in those who had smaller hip circumferences than expected from their BMI. Adjusted odds ratios (95% CI) for NIDDM were 2.9 (1.6-5.1) in men and 2.8 (1.5-5.1) in women who had a larger than expected waist, and were 3.7 (2.1-6.5) in men and 2.1 (1.1-3.8) in women who had smaller than expected hips. CONCLUSIONS: Our findings suggest that diabetics have larger waists and smaller hips compared to nondiabetics, irrespective of their degree of obesity, age and life-style factors. One possibility is that besides abdominal fatness, peripheral muscle atrophy is one of the factors associated with NIDDM. PMID- 9408071 TI - The effect of metformin on blood pressure and metabolism in nondiabetic hypertensive patients. AB - OBJECTIVES: To study the effect of metformin on blood pressure and metabolism in nondiabetic hypertensives. DESIGN: A six-week single-blind placebo wash-out, followed by a double-blind placebo-controlled parallel group design with skew randomization (2:2:1) to metformin 850 mg b.i.d. (n = 10), metformin 500 mg b.i.d. (n = 10), or placebo b.i.d. (n = 5) for 12 weeks. Office blood pressure (oBP), ambulatory blood pressure (aBP), lipoproteins, and oral glucose tolerance (OGTT) were measured/performed before and during treatment. SUBJECTS: Sixteen male and nine female nondiabetic (OGTT) patients (median age 57 (39-74) years) with verified hypertension (White-coat excluded) for 4 (0-20) years. RESULTS: The possible effect of metformin treatment and dosage was tested with a two-factor analysis of variance. Treatment induced a significant decline in diastolic oBP, P < 0.05. This decline was, however, not significantly different comparing metformin and placebo. Systolic oBP, diastolic aBP, and systolic aBP showed no significant change by treatment. The decline in diastolic oBP was 5 mmHg in the pooled group of metformin-treated patients, P < 0.005. Different gender and the presence of obesity had no impact on the decline in diastolic oBP within this group. Changes in fasting C-peptide and fasting insulin during treatment were unrelated to blood pressure changes. High fasting insulin (> 60 pmol L[-1]) or high fasting C-peptide (> 1000 pmol L[-1]) at baseline did not favour an effect of metformin on diastolic oBP. Glucose metabolism and lipoproteins were unchanged in all groups. CONCLUSIONS: Although metformin treatment induced a decline in diastolic office blood pressure in nondiabetic hypertensives, the decline was not different from that during placebo treatment. Metformin had no significant effect on ambulatory blood pressure. Thus, metformin has, if any, only a minor clinically insignificant effect on blood pressure in nondiabetic hypertensives. The study does not support the hypothesis that circulating insulin is a major regulator of blood pressure in hypertension. PMID- 9408072 TI - Colonic endocrine cells in inflammatory bowel disease. AB - OBJECTIVES: To study colonic endocrine cell types in patients with ulcerative colitis (UC) and Crohn's disease (CD). SETTING: Departments of Medicine and Pathology, University Hospitals, Umea and Uppsala, Sweden. SUBJECTS: Seventeen patients with UC (seven females and 10 males) and 11 patients with CD (five females and six males). Twenty-two patients (eight females and 14 males) operated on for colon carcinoma and without signs of inflammatory bowel disease were used as controls. MEASUREMENTS: The colonic endocrine cell types were identified by immunohistochemical methods and quantified by computed image analysis. RESULTS: The areas of the argyrophil cells as well as those immunoreactive to chromogranin A and serotonin were significantly increased in patients with both UC and CD, compared with those in the controls. In patients with CD, the areas of polypeptide YY(PYY)- and pancreatic polypeptide (PP)-immunoreactive cells were significantly reduced, whilst the area of enteroglucagon-immunoreactive cells was increased. There was no statistical difference in endocrine cell area between specimens with slight versus severe inflammation, except for PYY and enteroglucagon cell areas in patients with CD. Whilst the former cell area decreased, the latter increased in specimens with severe inflammation. The mean cellular area for each endocrine cell type did not differ between the controls and patients with UC or CD. CONCLUSIONS: The increase in the serotonin immunoreactive cell area in patients with both UC and CD might be one of the factors causing reduced colonic contraction and increased intraluminal pressure observed in patients with inflammatory bowel disease. Furthermore, in patients with CD, the decreased PYY-immunoreactive cell area may explain the decreased absorption and increased secretion found in these patients. PMID- 9408073 TI - Schizophrenic symptoms and SPECT abnormalities in a coeliac patient: regression after a gluten-free diet. AB - A 33-year-old patient, with pre-existing diagnosis of 'schizophrenic' disorder, came to our observation for severe diarrhoea and weight loss. Use of single photon emission computed tomography, (99mTc)HMPAO SPECT, demonstrated hypoperfusion of the left frontal brain area, without evidence of structural cerebral abnormalities. Jejunal biopsy showed villous atrophy. Antiendomysial antibodies were present. A gluten-free diet was started, resulting in a disappearence of psychiatric symptoms, and normalization of histological duodenal findings and of (99mTc)HMPAO SPECT pattern. This is the first case in which, in an undiagnosed and untreated coeliac patient with psychiatric manifestations, the (99mTc)HMPAO SPECT demonstrated a dysfunction of frontal cortex disappearing after a gluten-free diet. PMID- 9408074 TI - Myelodysplastic syndrome with vasculitic manifestations. AB - Vasculitis in patients with the myelodysplastic syndrome (MDS) is a rare phenomenon. We describe a patient who presented with necrotic lesions of his toes, which proved to be the result of immune complex mediated vasculitis. This unusual combination of vasculitis and MDS prompted us to review the literature. Forty-four cases of vasculitis in association with MDS were found. The pathogenesis of the vasculitis in MDS remains speculative, although several reports suggest an immunological mechanism. The temporal relationship could not be determined in 20 (45%) of reported cases. The prognosis of these patients appears to be worse than in patients with MDS without vasculitis. Steroids were used in 41 (93%) of the reported cases. Our patient did not receive any drug therapy, nevertheless his necrotic lesions improved within a few weeks. PMID- 9408075 TI - Limited Wegener's granulomatosis with central nervous system involvement and fatal outcome. AB - A 74-year-old woman was admitted to the medical department with vertigo and confusion, and suffered seizures three days later. Clinical, radiological and histopathological examination revealed limited Wegener's granulomatosis (WG) with central nervous system (CNS) involvement. Initially she was treated successfully with prednisolone and cyclophosphamide, but she relapsed after 25 months and died four weeks later. Autopsy showed widespread granulomas with vasculitis in the cerebrum. These findings emphasize the importance of considering WG in CNS disease of unclear origin, and demonstrate the aggressive nature of the disease. PMID- 9408076 TI - Sustained trilineage response to erythropoietin therapy in a case of aplastic anaemia. PMID- 9408077 TI - Localization of functional domains in the androgen receptor. AB - Functional domains of the androgen receptor (AR) have been localized through a combination of studies on naturally occurring AR gene mutations, in vitro mutagenesis studies and comparison with the structure of other members of the steroid/nuclear receptor superfamily. Two activation domains exist within the amino-terminal domain, and a ligand-dependent activation domain is present in the ligand binding domain. The poly(Gln) stretch within the amino-terminal domain may inhibit the transactivation function of the receptor. Different ligands or binding to different promoters may recruit the use of different activation domains, which may provide promoter-specific effects of receptor action. Co activator proteins that modulate or enhance AR action have been identified, many of which interact with the ligand binding domain of the AR. Tissue-specific expression of such co-activators, and promoter-specific protein interactions, may also help control the specificity of androgen action. Target Ser residues for phosphorylation have been identified, which may be the site of action for cross talk from protein kinase signalling pathways. However, the role of phosphorylation in AR function in general is still unclear. It is now clear that interactions occur between receptor domains, modulating functions including ligand dissociation, dimerization and transactivation. By studying the functional domains of the AR, and how they control receptor function in response to different activation signals, we are beginning to understand the mechanisms controlling the specificity of receptor action. PMID- 9408078 TI - A sequence in the 5' flanking region confers progestin responsiveness on the human c-myc gene. AB - Previous reports have shown that progestins stimulate the proliferation of the human breast cancer cell line T47D in culture. Under different conditions other reports have shown progestin stimulation, inhibition or no effect on growth. It has also been shown that c-myc expression is stimulated at early times by progestins. We are currently testing the hypothesis that the mechanism of growth enhancement by progestins involves the stimulation of expression of c-myc. This hypothesis predicts a progesterone regulatory region in or near the c-myc gene. We have identified a region, from -2327 to -1833, which serves this function. This region includes a 15 bp sequence with homology to the PRE (progesterone response element) consensus sequence. Human progesterone receptor (PR) binds to this sequence in a specific, ligand-enhanced manner in electrophoretic mobility shift assays (EMSA). A 3507 bp HindIII-XbaI fragment of the 5' flanking region of the c-myc gene, -2327 to +1180, containing the progestin regulatory region and the c-myc promoter, confers progestin responsiveness to the CAT (chloramphenicol acetyl transferase) reporter gene in progesterone receptor (PR)-rich T47D human breast cancer cells, but not in PR-negative MDA-MB-231 cells. Removal of the progestin regulatory region abrogates progestin responsiveness. These data demonstrate that the sequence from -2327 to -1833 of the human c-myc gene includes a positive progestin regulatory region. PMID- 9408079 TI - The mammalian metabolite, 2-methoxyestradiol, affects P53 levels and apoptosis induction in transformed cells but not in normal cells. AB - The endogenous metabolite, 2-methoxyestradiol (2ME), is an inhibitor of tubulin polymerization and is therefore toxic to dividing fast-growing tumor cells. Transformed cells are not equally susceptible to the effects of 2ME. In this study the effects of 1-2 microM doses of 2ME on cell cycle progression, apoptosis induction and on p53 levels were evaluated using flow cytometry in cells with different p53 status. No effect of 2ME was seen in normal human skin fibroblast strain HSF43 with wild-type (wt) p53. However, in SV40 T antigen transformed HSF43 cells (line E8T4), 2ME caused a prominent G2/M arrest, with subsequent micronuclei formation followed by apoptosis. Increased p53 levels were present in the G2/M cells. Our results suggest that 2ME, being a microtubule poison, may release the bound p53 from T antigen, and that this p53 may enhance the apoptotic effects. Two lymphoblast cell lines derived from the same donor, TK6, expressing low levels of wt p53, and WTK1, expressing high levels of mutant p53, showed similar moderate responses to 2ME at 37 degrees C. The effects included enhanced apoptosis and a modest G2/M block. No increase in p53 levels was seen. However, at the permissive temperature of 30 degrees C marked increases in apoptosis and a prominent G2/M-phase block, similar to that seen in the E8T4 cells, were present in the WTK1 cells, indicating that the high levels of mutant p53 have now become functional, enhancing the apoptotic effects initiated by 2ME. PMID- 9408080 TI - Protein synthesis is not implicated in the ligand-dependent activation of the estrogen receptor in MCF-7 cells. AB - In MCF-7 cells, estrogen receptor (ER) elimination occurs rapidly under stimulation with estradiol (E2) at 1 nM ('ER processing'); cycloheximide (CHX) at 50 microM impedes this phenomenon. ER processing is also observed when E2 is removed after the first hour of incubation, indicating that the role of the hormone would be limited to the initiation of this process. When CHX is removed at the same time, receptor processing and, later, the induction of progesterone receptor (PgR) both proceed. The initial estrogenic signal which activates ER is therefore not influenced by CHX. In support of this conclusion, no effect of the drug on E2 binding affinity of residual ER was detected. A similar result was recorded for a series of estrogens and antiestrogens, indicating that CHX exerts no influence on the potential agonistic/antagonistic potency of any ligand. Size exclusion chromatography (FPLC) revealed that [3H]E2-induced ER activation leads to the cleavage of the native receptor (67 kDa) into low molecular weight isoforms which subsequently become less detectable over time (proteolysis). In the presence of CHX, such ER isoforms persist, confirming the absence of interference of the drug with the activation step. When the cells were prelabelled with [3H]tamoxifen aziridine ([3H]TAZ) before their exposure to E2, ER cleavage could not be detected due to the lack of activation potency of the antiestrogenic ligand. However, the [3H]TAZ-ER complexes were subjected to E2 induced processing; CHX blocked this phenomenon, which is associated with the maintenance of ER synthesis and activation. PMID- 9408081 TI - Cloning of two alternatively spliced 21-hydroxylase CDNAs from rat adrenal. AB - Interest in extra-adrenal corticosteroid synthesis has been revived by technological advances and the quest for answers to clinical problems. The cytochrome P450 21-hydroxylase converts progesterone to deoxycorticosterone, the obligatory substrate for the production of the main adrenal steroids aldosterone, cortisol and corticosterone. The rat P450 21-hydroxylase was cloned and two constructs, 21OH-5 and 21OH-6, sequenced. The constructs are similar, except that 21OH-6 has three additional major insertions of 64, 70 and 84 bp, a 3 bp deletion, and four extra base pairs immediately before the poly-A sequence. The entire coding region of 21OH-5 has 87 and 71% homology with the mouse and human 21-hydroxylase cDNA, respectively, whereas the encoded protein has 84 and 65% homology. Reverse transcriptase-polymerase chain reaction (RT-PCR) combined with Southern blot demonstrated expression of both transcripts in the kidney, aorta, liver, cerebellum, hypothalamus and brain stem, heart and cerebrum, but not the hippocampus, in addition to the adrenal. The entire coding region of 21OH-5 and the corresponding region of 21OH-6 including the three introns were cloned into pCR3 and the plasmids transiently transfected into COS-7 cells. Only 21OH-5 was translated into active protein, converting approximately 64% of 3H-progesterone to deoxycorticosterone in 2 h. PMID- 9408082 TI - An N-terminally truncated third progesterone receptor protein, PR(C), forms heterodimers with PR(B) but interferes in PR(B)-DNA binding. AB - Multiple progesterone receptor (PR) isoforms may explain in part the complex and diverse biological actions of progestins. Recent studies demonstrate that the human 116 kDa B-receptor (PR[B]) and the 94 kDa A-receptor (PR[A]) can have very different transcriptional functions that are cell- and promoter-specific. Additionally, we have shown the existence of a smaller N-terminally truncated 60 kDa progestin-specific binding protein, called the C-receptor (PR[C]), that has unique transcriptional potentiating properties. In the presence of the other two PR isoforms, PR(C) enhances the transcriptional activities of the larger PR proteins. In order to determine the mechanism of action for the transcriptional promoting abilities of PR(C), the structural and functional properties of PR(C) were analysed and compared to those of PR(A) and PR(B). PR(C) consistently displayed a dissociation constant (Kd) approximately 5 times higher than that for PR(B) and PR(A), suggesting that the N-terminal truncation of PR(C) results in a conformation different from the two larger PR isoforms, that affects the hormone binding region and its interaction with hormone. Despite this change, PR(C) is still capable of forming heterodimers with the larger PR(B) in solution, as determined by co-immunoprecipitation studies, but PR(C) interferes in tight PR(B) binding to DNA in gel-shift assays. Surprisingly, progestin and antiprogestin autoregulation of PR(C) protein levels parallel those for PR(B) and PR(A). PMID- 9408083 TI - The regional brain distribution of the neurosteroids pregnenolone and pregnenolone sulfate following intravenous infusion. AB - We have studied the distribution of the neurosteroids pregnenolone (Pe) and pregnenolone sulfate (PeS) in seven brain regions, and plasma and fat tissues in male adult rats following the intravenous infusion of 14 mg/kg Pe and 18 mg/kg PeS, respectively. After chromatographic separation of steroid sulfate esters and non-conjugated steroids by solid phase octadecyl C18 columns and celite column chromatographic separation of Pe from cross-reacted steroids, the concentrations of Pe and PeS were determined by radioimmunoassay. We found that both Pe and PeS concentrations were significantly increased in plasma, fat and brain compared to the vehicle controls after i.v. infusion of Pe and PeS. In the controls, Pe concentrations were highly correlated within brain regions and between fat and brain regions. Most correlations were lost after Pe and PeS infusions. The content of Pe and PeS was not uniformly distributed in the brain. The hypothalamus contained the highest level of Pe in controls, Pe-infused and PeS infused rats (12 +/- 3.1, 3500 +/- 180 and 590 +/- 54 ng/g, respectively). The highest concentration of PeS was detected in the hypothalamus (26 +/- 8.2 ng/g) and striatum (17 +/- 4.1 ng/g) in controls, in the hypothalamus (200 +/- 24 ng/g) after PeS infusion as well as in the hypothalamus and medulla oblongata (57 +/- 9.6 and 55 +/- 7.6 ng/g, respectively) after Pe infusion. This study has yielded evidence that PeS injected i.v. can cross the blood-brain barrier without being hydrolysed to the more lipophilic Pe, and can thus be taken up by the brain. PMID- 9408084 TI - Transcriptional regulation by HNF-4 of the steroid 15alpha-hydroxylase P450 (Cyp2a-4) gene in mouse liver. AB - The mouse P450 gene Cyp2a-4 encodes the hepatic steroid 15apha-hydroxylase. We have defined in the 5'-flanking sequence of Cyp2a-4 gene, a composite regulatory element (-61AGACCAAAGTCCGGCCTTC-42) which contains a potential CpG methylation site at position -50. Gel-shift assays indicate that this element consists of overlapped binding sites for a hepatocyte-enriched transcription factor HNF-4 and a Sp1-like protein. Moreover, transcription of the Cyp2a-4 gene is activated by coexpression of HNF-4 in HepG2 cells. A mutation (C at -50 to A) abolishes the binding of HNF-4 to the element as well as the transcriptional activation by HNF 4. The methylated C at position -50, however, does not affect HNF-4 binding. Neither the mutation nor the methylation at position -50 affect the binding of Sp1-like protein to the element. It appears, therefore, that HNF-4 activates the hepatic transcription of Cyp2a-4 gene through its direct binding to the regulatory element regardless of the methylation at position -50. PMID- 9408085 TI - Distribution and characterization of neurosteroid sulfatase from the bovine brain. AB - We investigated the regional and subcellular distribution of neurosteroid sulfatase (NSS) in the bovine brain and its enzymatic properties by using dehydroepiandrosterone sulfate (DHEA-S) as a substrate. Bovine NSS was highly concentrated in the region of the midbrain and in the hypothalamus. The enzyme was found to be a microsomal enzyme. The optimal temperature of the enzyme was 50 degrees C, which was slightly lower than that of other steroid sulfatases. The optimal pH of bovine NSS was 7.4 with a second optimum at pH 4.0. The second optimal pH of 4.0 was the most characteristic property of bovine NSS. Employing DHEA-S as the substrate, apparent Km and Vmax values were 113 +/- 21 microM and 4.1 +/- 0.4 nmol/mg protein/h, respectively, whereas Km and Vmax values were found to be 1.6 +/- 0.2 M and 1.9 +/- 0.3 micromol/mg protein/h with p nitrophenyl sulfate (NP-S) as the substrate. NSS has thus been shown to have a higher affinity for the steroid sulfate than the phenolic compound. When DHEA-S was used as the substrate, pregnenolone sulfate (Preg-S) was a competitive inhibitor with an apparent Ki value of 46 microM, and NP-S was a non-competitive inhibitor (apparent Ki=12 mM). PMID- 9408086 TI - Effects of danazol and progesterone on sex hormone-binding globulin mRNA expression in human endometrial cancer cell line Ishikawa. AB - To ascertain one of the biological effects of danazol and progesterone on the uterine endometrial cancer cell line, Ishikawa, we investigated the effects of these steroids on sex hormone-binding globulin (SHBG) mRNA expression by competitive reverse transcription-polymerase chain reaction-Southern blot analysis (RT-PCR-SBA). Estradiol-17beta (E2) in any concentration given did not exert any significant effect on the expression of SHBG mRNA. Danazol and progesterone significantly (P < 0.05) suppressed the expression of SHBG mRNA dose dependently starting at a concentration of 10(-6) and 10(-8) M, respectively. Progesterone, in a low concentration (10[-10] M) with E2 (10[-8] M), significantly (P < 0.05) increased the expression of SHBG mRNA, but danazol did not. In contrast, danazol and progesterone in high concentrations (10[-6] to 10[ 5] M) with E2 (10[-8] M) significantly (P < 0.05) suppressed its expression. The time course study showed the time-dependent decrease of SHBG mRNA level by danazol and progesterone (10[-6] M) with or without E2 (10[-8] M), except for a temporal increase by progesterone. These findings suggest that danazol and progesterone in a superphysiological milieu down-regulate the intracellular SHBG related steroidal actions, and that progesterone in a physiological milieu with estrogen up-regulates it in a hormone-dependent cell line. A decrease of intracellular SHBG caused by high-dose danazol or progesterone might partly contribute to the abolition of the intracellular estrogen-dominant milieu, and be related to the inhibition of estrogen-dependent growth of some endometrial cancer cells. PMID- 9408087 TI - Demonstration of membrane estrogen binding proteins in rat brain by ligand blotting using a 17beta-estradiol-[125I]bovine serum albumin conjugate. AB - This paper describes a ligand blotting procedure to visualize membrane estrogen receptors/binding proteins immobilized on nitrocellulose membranes. Using 17beta estradiol covalently linked with [125I]bovine serum albumin (BSA) at the C-6 position (17beta-E-6-[125I]BSA) as a ligand, three major binding proteins with molecular masses of approximately 23, 28, and 32 kDa were identified from crude synaptosomal fractions (P2) of female rat brains. The binding of 17beta-E-6 [125I]BSA to these proteins is selective for 17beta-estradiol because BSA had no effect, and 17alpha-E-6-BSA was at least two orders of magnitude less potent than 17beta-E-6-BSA in displacing the binding. In addition, [125I]BSA and 17alpha-E-6 [125I]BSA at similar concentrations did not bind to these proteins. Competition and saturation assays indicate that the binding affinity of 17beta-E-6-[125I]BSA for these proteins was in the range of 1-10 nM. These proteins are not contaminants from cytosolic or serum estrogen binding proteins since no corresponding protein bands were found in cytosolic fractions. Three additional protein bands with molecular masses of approximately 18, 40, and 130kDa were also detected, although inconsistently. The 23 and 40 kDa proteins seem to be concentrated in mitochondrial fractions (mP2), whereas the 28 and 32 kDa proteins are enriched in microsomal fractions (P3). Application of digitonin-solubilized P2 fractions to 17beta-estradiol-coupled affinity columns resulted in significant purification of the 23 kDa protein as shown by ligand blotting. This protein was later identified as oligomycin-sensitivity conferring protein (OSCP), as reported previously. These data indicate that specific estrogen binding proteins different from classical nuclear estrogen receptor (66 kDa) are present in the cellular membranes of the female rat brain. The ligand blotting technique described here would also be applicable for the identification of other membrane steroid binding proteins/receptors using similar radiolabelled steroid-BSA conjugates. PMID- 9408088 TI - Cortisol metabolism and its inhibition by glycyrrhetinic acid in the isolated perfused human placental lobule. AB - We have previously reported the placental metabolism of prednisolone to prednisone, 20alpha- and beta-dihydroprednisone and 20beta-dihydroprednisolone. In this study, the disposition of cortisol was investigated in vitro in the dual perfused, isolated human placental lobule after the addition of cortisol (1.2 micromol, n = 3 and 12 micromol, n = 4) to the maternal compartment. Analysis of 5 h maternal and fetal perfusate samples by high performance liquid chromatography-electrospray-tandem mass spectrometry (HPLC-ESI-MS/MS) revealed that cortisol was mainly metabolized to cortisone, but a significant production of 20alpha-dihydrocortisone, 20beta-dihydrocortisone, 20alpha-dihydrocortisol and 20beta-dihydrocortisol was also detected. Saturability of metabolism but not transfer was demonstrated. Metabolism was eliminated by co-perfusion with the potent 11beta-hydroxysteroid dehydrogenase (11beta-HSD) enzyme inhibitor 18beta glycyrrhetinic acid (GA). The disposition of GA was analysed using HPLC atmospheric pressure chemical ionisation-MS/MS (HPLC-APCI-MS/MS). GA was found to transfer from the maternal to the fetal circulations without detectable metabolism during 6 h of perfusion. PMID- 9408089 TI - Dexamethasone metabolism in vitro: species differences. AB - Dexamethasone (DEX) is extensively metabolized to 6-hydroxyDEX (6OH-DEX) and side chain cleaved metabolites in human liver both in vitro and in vivo with CYP3A4 responsible for the formation of 6-hydroxylated products. In the present study, the metabolism of [3H]DEX has been examined in the liver fractions from various mammalian species and metabolite profiles compared with those obtained with human liver microsomes. Metabolites were quantified by radiometric high-pressure liquid chromatography (HPLC) and characterized by liquid chromatography-mass spectrometry (LC-MS) and co-chromatography with chemical standards, where available. 6OH-DEX formation was quantified for each species and the inhibitory potency of ketoconazole at 1 and 20 microM determined. Glycyrrhetinic acid, a specific inhibitor of 11-dehydrogenase, was also used to determine the extent of reductive DEX metabolism. Species differences in metabolite profiles obtained from microsomal incubations were both quantitative and qualitative. 6 Hydroxylation was variable (highest in the hamster) and was not always the major route of metabolism, and formation was sex-specific in the rat (male >> female). The inhibition of 6-hydroxylation (CYP3A) by ketoconazole was variable, and indicates that ketoconazole cannot be regarded as a selective inhibitor of CYP3A proteins in all species. Cytosolic incubations produced similar profiles in different species with the formation of a metabolite (M5) which was inhibited by glycyrrhetinic acid and tentatively identified in this study as 11-dehydro-side chain cleaved DEX (11DH-9alphaF-A). In conclusion, the male rat gave a metabolite profile which was closest to that seen in the human. However, 6-hydroxylation was most extensive in the hamster which may therefore be a suitable model to use for further studies on DEX metabolism by CYP3A. PMID- 9408090 TI - Drosophila ecdysone receptor functions as a constitutive activator in yeast. AB - Transcriptional activation of the Drosophila ecdysone receptor (EcR) was studied in yeast cells, which carry a reporter plasmid containing the ecdysone response element in the absence or presence of its heterodimeric partners, ultraspiracle protein (USP) or human retinoid X receptor (RXRalpha). High constitutive transcriptional activation was detected in the yeast strain expressing EcR, but not USP or RXRalpha in the absence of ponasterone or muristerone A. Incubation of these ligands with yeast cells coexpressing EcR and USP or RXRalpha did not enhance the constitutive transcriptional activity. However, specific ligand binding using [3H]ponasterone A as a radioactive ligand was detected only in yeast extracts prepared from the yeast strain coexpressing EcR and USP, but not from yeast strains expressing only EcR or USP. The ligand binding characteristics of the EcR/USP complexes were similar to those reported in an insect cell line with a Kd value of 1.8 nM for [3H]ponasterone A. These data are in contrast to mammalian cell transfection studies, and indicate that the EcR is the only member of the nuclear receptor superfamily of ligand-activated transcription factors which functions as a constitutive transcriptional activator in yeast, although the EcR/USP complexes exhibit normal ligand binding properties. PMID- 9408091 TI - Clinical features, investigation, and management of patients with defects of mitochondrial DNA. PMID- 9408092 TI - Neurological picture. Bilateral internuclear ophthalmoplegia in multiple sclerosis. PMID- 9408093 TI - Age at immigration to England of Asian and Caribbean immigrants and the risk of developing multiple sclerosis. AB - OBJECTIVES: Previous studies have shown that multiple sclerosis is very uncommon among Indian and Pakistani immigrants to England but that their children born in the United Kingdom, in the age groups available for study, have a similar risk of developing the disease as occurs in the general British population. The present study was to ascertain if these immigrants who enter England as children below the age of 15, have a higher risk of developing multiple sclerosis than those that enter after this age. METHODS: A search was made in Greater London, the West Midlands, Leicester, Bradford, Halifax, and Huddersfield to find ethnic Indian, Pakistani, and Bangladeshi immigrants to England with multiple sclerosis. During the course of the study some immigrants from the Caribbean with multiple sclerosis were also found. The population at risk by ethnic group and age at entry was not available from the 1991 Census but was available in the annual Labour Force Surveys. RESULTS: Indian and Pakistani immigrants who entered England younger than 15 had a higher risk of developing multiple sclerosis than those that entered after this age. Caribbean immigrants, who have a higher multiple sclerosis prevalence than Asian immigrants, did not show this difference. CONCLUSION: This study confirms previous studies which show that the environment during childhood is a major factor in determining the risk of developing multiple sclerosis. PMID- 9408095 TI - Physiological effects of selective tibial neurotomy on lower limb spasticity. AB - OBJECTIVES: To assess by electrophysiology the effect of tibial selective neurotomy on muscle imbalance of the spastic ankle. METHOD: The amplitudes of the H reflexes, M responses (muscle contractions recorded after stimulation of the tibial nerve), and Hmax:Mmax ratio were recorded in 12 patients with chronic lower limb spasticity, before and one month after tibial selective neurotomy. Recordings were done on medial and lateral gastrocnemius and soleus muscles. Clinical evaluation was done with both global (Held's score) and analytical tests (step measurements, gait velocity, and ankle angulation during active and passive movements). RESULTS: After neurotomy, gait improved in all patients. Held's score of spasticity was better in all patients. Active dorsiflexion of the ankle was unchanged in three patients, but the others improved by 5 degrees to 12 degrees. Hmax, Mmax, and Hmax:Mmax ratios were lower. The Hmax on the gastrocnemius muscle, clinical strength, Mmax of all the muscles, and Hmax:Mmax ratio for the soleus and lateral gastrocnemius muscle were significantly lower after surgery. CONCLUSION: There was an improvement of clinical and electrophysiological spastic indices after selective tibial neurotomy. Neurotomy acted not only on motor neurons by decreasing strength, but also the reflex enlargement by decreasing sensory afferents. PMID- 9408094 TI - Evaluation of malignancy in ring enhancing brain lesions on CT by thallium-201 SPECT. AB - OBJECTIVE: To investigate patients with cystic enhancing lesions on CT and to determine whether thallium-201 (201Tl) SPECT adds to further preoperative information in differential diagnosis between gliomas and abscesses. METHODS: Twenty one patients with cystic ring enhancing CT findings were studied and uptake indices were compared with CT enhancement volumes, histopathology, and survival times. RESULTS: Fourteen high grade gliomas, three low grade gliomas, and four abscesses were found. Uptake was higher in the highly malignant glioma group (median thallium index (TI)=2.1), than in the low grade glioma group (median TI=1.4) or among the abscesses (median TI=1.6). Overlapping indices were found between high and low malignant cystic gliomas as well as between either one of the glioma groups and the infectious lesions, and there were no significant differences between groups. There was a level at the value 2, where TI > or = 2 correlated with tumour diagnosis. One low grade tumour had an extremely high index and a very high enhancement volume. Indices correlated significantly with CT enhancement volumes (P=0.005). There was no significant correlation between Tl indices and patient survival times among the high grade gliomas. One patient with a highly malignant tumour but low Tl uptake < 2, had a survival > five years. CONCLUSIONS: It is concluded that high 201Tl uptake in enhancing cystic lesions is an indicator of highly malignant glioma. However, the differentiation between the high malignant gliomas and abscesses or low malignant gliomas by 201TL SPECT is only partial with an overlap between these groups. PMID- 9408096 TI - Response of parkinsonian swallowing dysfunction to dopaminergic stimulation. AB - OBJECTIVES: To determine the degree of dopaminergic response of swallowing dysfunction in Parkinson's disease. METHODS: Fifteen patients with idiopathic Parkinson's disease and symptomatic dysphagia were studied. All had motor fluctuations in response to long term levodopa therapy. On two separate days, after overnight withdrawal of all antiparkinsonian medication, a modified barium swallow using cinefluoroscopy and different food consistencies was performed before and after administration of oral levodopa and subcutaneous apomorphine. RESULTS: Despite all patients having an unequivocal motor response to both agents, there were few significant responses in any of the quantitative or qualitative criteria of swallowing dysfunction assessed. The oral preparatory phase, generally considered a more voluntary component of swallowing, showed a response, but not with all consistencies. In a subgroup of patients the pharyngeal phase time also improved. CONCLUSIONS: These findings suggest that parkinsonian swallowing dysfunction is not solely related to nigrostriatal dopamine deficiency and may be due to an additional non-dopamine related disturbance of the central pattern generator for swallowing in the pedunculopontine nucleus or related structures in the medulla. PMID- 9408097 TI - Prevalence of orthostatic hypotension in Parkinson's disease. AB - OBJECTIVES: To investigate the prevalence of orthostatic hypotension and the nature of the postural events related to a fall in blood pressure in patients with Parkinson's disease. METHODS: Blood pressure was measured first in a supine position after a rest of at least 15 minutes and every minute during 10 minutes of an active standing up procedure. Orthostatic hypotension was considered as present when a fall of at least 20 mm Hg of systolic blood pressure was recorded. Postural events which occurred during the standing test were identified from a questionnaire and self reporting. Statistical analysis was performed to determine the relation between orthostatic hypotension and disease characteristics (duration, severity) and the use of antiparkinsonian drugs. Ninety one consecutive patients with Parkinson's disease (48 women, 43 men, mean age 66 (SD 9) years) participated to the study. RESULTS: A fall of at least 20 mm Hg of systolic blood pressure was found in 58.2% of the patients. Orthostatic hypotension was asymptomatic in 38.5% and associated with postural events in 19.8% of the patients. Symptomatic (but not asymptomatic) orthostatic hypotension was related to duration and severity of the disease and with the use of higher daily levodopa and bromocriptine doses. The analysis of the relation between the postural symptoms (and the need for standing test abortion) with the fall in systolic blood pressure allowed the identification of six clinical criteria specific of orthostatic hypotension. A direct relation between the postural changes in systolic blood pressure and the number of clinical events in this clinical scale was found. CONCLUSION: The frequency of orthostatic hypotension in Parkinson's disease is high and it is possible to establish a clinical rating scale which could be used to assess the effects of drugs employed in the management of orthostatic hypotension. PMID- 9408099 TI - Paul Ferdinand Gachet (1828-1909). PMID- 9408098 TI - Parkinson's disease and depression: evidence for an alteration of the basal limbic system detected by transcranial sonography. AB - OBJECTIVES: Depression is a frequent symptom in Parkinson's disease. Compelling evidence suggests a role of the brainstem in the control of mood and cognition. In patients with unipolar depression transcranial sonography (TS) studies have shown structural alteration of the mesencephalic brainstem raphe which could suggest an involvement of the basal limbic system in the pathogenesis of primary mood disorders. The objective of the present study was to evaluate whether a similar alteration could be found in depressed patients with Parkinson's disease using TS. METHODS: Thirty patients with Parkinson's disease and 30 age and sex adjusted controls were examined by TS. Raphe echogenicity was rated semiquantitatively. The severity of motor symptoms and depression was rated using standard research instruments. RESULTS: Raphe echogenicity was significantly reduced in depressed patients with Parkinson's disease compared with nondepressed patients with Parkinson's disease and control subjects. Raphe echogenicity correlated negatively with degree of motor impairment, and differences in raphe echo between depressed and non-depressed patients with Parkinson's disease were upheld when motor impairment was controlled for. CONCLUSION: These preliminary findings suggest that, as in unipolar depression, a morphological alteration of the brainstem raphe might be involved in the pathogenesis of depression in Parkinson's disease. This raphe alteration may reflect involvement in the basal limbic system in the pathogenesis of secondary depression. This concept is in line with current knowledge on the pathogenesis of both depression in Parkinson's disease and primary depressive disorders. PMID- 9408101 TI - Changes in cerebral blood flow and vasoreactivity in response to acetazolamide in patients with transient global amnesia. AB - OBJECTIVE: Previous reports about changes in cerebral blood flow (CBF) in transient global amnesia disclosed decreased flow in some parts of the brain. However, CBF analyses in most reports were qualitative but not quantitative. The purpose of this study was to determine changes in CBF in transient global amnesia. METHODS: The CBF was measured and the vasoreactive response to acetazolamide was evaluated in six patients with transient global amnesia using technetium-99m hexamethylpropylene amine oxime single-photon emission computed tomography (SPECT). The CBF was measured during an attack in two patients and soon after an attack in the other four. About one month later, CBF was re evaluated in each patient. RESULTS: Two patients examined during an attack and one patient examined five hours after an attack had increased blood flow in the occipital cortex and cerebellum. Three patients examined at six to 10 hours after an attack had decreased blood flow in the thalamus, cerebellum, or putamen. These abnormalities of blood flow almost disappeared in all patients one month after onset. The vasodilatory response to acetazolamide, which was evaluated initially using SPECT, was poor in areas of increased blood flow. By the second evaluation of CBF with acetazolamide, the vasodilatory response had returned to normal. CONCLUSIONS: In a patient with transient global amnesia, CBF increased in the vertebrobasilar territory during the attack and decreased afterwards. The vasodilatory response to acetazolamide may be impaired in the parts of the brain with increased blood flow. It is suggested that transient global amnesia is distinct from migraine but may share the same underlying mechanism. PMID- 9408100 TI - Temporal lobe abnormalities in dementia and depression: a study using high resolution single photon emission tomography and magnetic resonance imaging. AB - OBJECTIVES: Perfusion SPECT and MRI were used to test the hypothesis that late onset depression is associated with brain abnormalities. METHODS: Forty depressed patients (DSM-III-R major depressive episode, not demented at two year follow up) were recruited who were either drug free, or on a stable dose of antidepressants for at least three weeks, as well as 22 demented patients (DSM-IIIR and NINCDS/ADRDA criteria for probable Alzheimer's disease). Patients were imaged at rest with a high resolution single slice 12 detector head scanner (SME-Neuro 900) and the cerebral perfusion marker 99mTc-exametazime (HM-PAO). Temporal lobe templates were fitted with brains pitched by 20 degrees-30 degrees. A subgroup of 41 patients (22 depressed) were also scanned using a Siemens Magnetron 1.0 Tesla magnetic resonance imager, using a FLAIR imaging sequence for the assessment of white matter hyperintensities, and a Turbo FLASH sequence for the measurement of medial temporal lobe width. RESULTS: Demented patients showed reduced perfusion, particularly in the left temporoparietal cortex. In these regions of interest, patients with late onset depression tended to have perfusion values intermediate between patients with early onset depression and demented patients. Differences in changes in white matter between demented and early and late onset depressive patients did not reach conventional levels of significance. Temporal lobe width differed between demented and depressed patients, but not between early and late onset depressed patients. Perfusion and temporal lobe width were not associated, but reductions of perfusion were associated with periventricular white matter changes. Mini mental state examination scores were associated with temporal perfusion in demented patients and with changes in deep white matter in depressed patients. Finally, severity of depressive symptoms was associated with decreased perfusion in frontotemporal and basal ganglia regions of interest. CONCLUSION: A cumulative effect of duration of illness on regional cerebral perfusion could not be confirmed. Late onset depression may show more abnormalities of deep white matter and of left temporoparietal perfusion than early onset depression, but the underlying pathology remains to be established. PMID- 9408102 TI - Unilateral lenticular infarcts: radiological and clinical syndromes, aetiology, and prognosis. AB - OBJECTIVES: To analyse the clinical features induced by lenticular infarction found in 20 patients, and to analyse the radiological and clinical correlations. METHODS: Eight women and 12 men, mean age 73 years, were included in this study, which was carried out from 1 January 1994 to 30 November 1996. They were characterised by the onset of a lenticular infarction, shown by CT and MRI. A complete neurological and neurocognitive examination, and photon emission computed tomography (SPECT), were performed in all the patients and there was a long clinical follow up. RESULTS: Two distinct clinical syndromes were identified corresponding to the two anatomical areas of the lenticular nucleus: behavioural and cognitive disorders were associated with infarcts within the globus pallidus, whereas both motor disorders (dystonia) and cognitive disorders were associated with infarcts within the putamen. Outcome was excellent in all the patients for motor function, but slight cognitive disorders, problems with short term memory, and dysphasia persisted for several months. The size of the lesion did not explain these symptoms. By contrast, the slight reduction in cerebral blood flow found in the adjacent frontotemporal area may explain them by a deafferentation or a diaschisis phenomenon. CONCLUSION: It is possible to identify the clinical symptoms of a single lesion in the pallidus nucleus and in the putaminal nucleus, in which behavioural, cognitive, and movements disorders are important. After an acute and spectacular onset, outcome is in general excellent. A disease of the small arteries must be involved. PMID- 9408103 TI - Early single photon emission computed tomography in Sturge-Weber syndrome. AB - OBJECTIVES: Functional cerebral imaging PET and SPECT have shown hypometabolism and hypoperfusion in the area of vascular malformation in children with epilepsy due to Sturge-Weber syndrome. However, data are scarce in infants and do not exist in patients with Sturge-Weber disease without epilepsy. The pattern of perfusion during the first two years of life was studied including patients before the onset of seizures. METHODS: Twenty two infants with later confirmed Sturge-Weber disease underwent SPECT examination using TOMOMATIC 564 (Medimatic) and xenon-133 at ages ranging from 8 days to 25 months. Twelve had never had seizures before SPECT and seven underwent a second SPECT a mean seven months later. Cerebral blood flow (CBF) was measured in the whole hemisphere and in the part of the cortex involved in the vascular malformation on both sides as well as a "pathological to normal" index for the hemisphere and vascular malformation. These values were compared with normal age paired values. RESULTS: Compared with controls, CBF and the indices in the hemisphere and vascular malformation were significantly decreased in patients who already had had seizures before SPECT, whereas they were significantly increased in 75% of the patients who had never had any seizures. On second SPECT, the indices were decreased in all patients, including the four who still remained non-epileptic. CONCLUSIONS: SPECT therefore detects CBF asymmetry in infants with Sturge-Weber disease, which tends to shift with age. The cortex involved in the vascular malformation is hyperperfused during the first year of life before first seizures. The classic hypoperfusion appears after one year of age, even in non-epileptic patients. PMID- 9408104 TI - Idiopathic generalised epilepsy in adults manifested by phantom absences, generalised tonic-clonic seizures, and frequent absence status. AB - OBJECTIVES: To describe the clinical and EEG features of adult patients with very mild absences, late onset generalised tonic clonic seizures, and frequent absence status. METHODS: Patients were referrals to a clinic for epilepsies. They all had clinical assessment and EEG, video EEG, or both for documentation of absences. RESULTS: Of 86 adults with idiopathic generalised epilepsies and EEG/video-EEG documented absences, 13 patients showed similar clinico-EEG features with: (a) "phantom absences" consisting of mild ictal impairment of cognition associated with brief (3-4 s), generalised 3-4 Hz spike/multiple spike and slow wave discharges; (b) infrequent, mainly late onset, generalised tonic clonic seizures, and (c), absence status which occurred in six of them either in isolation or terminating with generalised tonic clonic seizures. None of the patients had myoclonic jerks or photosensitivity. Two patients were father and daughter and another patient had a family history of infrequent generalised tonic clonic seizures. CONCLUSION: It seems that this is an idiopathic generalised epilepsy syndrome in adults which has not been previously recognised. PMID- 9408105 TI - Two families with autosomal recessive spastic paraplegia, pigmented maculopathy, and dementia. AB - OBJECTIVE: Two families with autosomal recessive hereditary spastic paraplegia and pigmented maculopathy are described. METHODS: All family members were examined by two neurologists. An assessment of cognitive function in affected members was made using the mini mental state examination (MMSE) or Cambridge cognitive examination (CAMCOG). RESULTS: Six patients from two families presented with a slowly progressive, autosomal recessive, spastic tetraplegia. Although they were always considered to be intellectually slower than their peers, further intellectual deterioration was noted during the second decade. Five had a pigmented maculopathy with mild decrease in visual acuity and all had distal amyotrophy, mild cerebellar signs, and developed faecal and urinary incontinence late in the course of the disease. CONCLUSION: The association of hereditary spastic paraplegia and pigmented maculopathy has rarely been described; only 11 families with 32 affected members have been reported, showing considerable heterogeneity in presentation. These described conditions may be allelic or more probably reflect mutations at different genetic loci. PMID- 9408106 TI - Eosinophilia-myalgia syndrome: selective cognitive impairment, longitudinal effects, and neuroimaging findings. AB - OBJECTIVE: To identify the specific nature of the neurocognitive impairments of eosinophilia-myalgia syndrome (EMS) in an unselected population, and to present longitudinal patterns. METHODS: A consecutive sample of 23 patients with EMS and 18 age and education matched control subjects were assessed on a comprehensive neuropsychological battery. Longitudinal results were gathered from six patients. RESULTS: Neurocognitive impairments were found which represent a subset of deficits reported in previous group and case study reports. Deficits were limited to complex visual memory, conceptual set shifting, and attention, which suggest a selective dysexecutive syndrome. The motor slowing and verbal memory deficits previously reported were not found. Although depression, fatigue, sleep deprivation, and pain were significant symptoms, they were unassociated with deficits with the exception of an association of depression with one deficit. There was no pattern of overall decline over time in a subset of the group, although considerable heterogeneity in the longitudinal patterns of neurocognitive tests was found. Abnormalities of white matter appeared in the MRI of eight of 12 patients. CONCLUSIONS: The neurocognitive and neuroimaging findings contribute to the evidence which indicates that the neural substrate of EMS is white matter damage. PMID- 9408107 TI - Symptomatic and functional outcome of surgical treatment of cervical dystonia. AB - OBJECTIVES: Previous studies of surgical treatment for cervical dystonia have reported highly variable rates of postoperative symptomatic benefit and morbidity. Little is known about functional improvement and long term results. This study evaluates the symptomatic and functional outcome of surgical treatment of cervical dystonia in a consecutive series of 46 patients. METHODS: The most affected muscles were selected for denervation after clinical examination and confirmation by four channel EMG studies. Surgical treatment, aiming at selective elimination of pathological activity while preserving normal motor function and avoiding side effects, was achieved by using a broad scope of techniques including intradural denervation, extradural denervation, and myotomy. Rather than carrying out standard operations, the treatment was tailored to the needs of the patient according to the individual pattern of dystonic activity. Long term benefit was assessed with a global outcome score, and a modified Toronto western spasmodic torticollis rating scale (TWSTRS) in those 34 patients who were available for a recent follow up evaluation. RESULTS: The 46 patients underwent a total of 70 procedures with intradural approaches in 33 instances, extradural approaches in 21, and muscle sections (singly or combined) in 22 instances. Transient mild postoperative side effects occurred in 10% of the procedures. The mean duration of long term follow up was 6.5 years. The global outcome was rated as excellent in nine patients (21%), as marked in 12 (27%), as moderate in nine (21%), as mild in nine (21%), and as no improvement in five (11%). A persistent side effect consisting of mild difficulty with balance was noted in one case. There were highly significant changes of the preoperative and postoperative mean values for almost all TWSTRS subscores for severity of cervical dystonia, functional disability, and pain. Patients with excellent outcome underwent a higher number of surgical procedures on average than those patients who achieved no benefit. CONCLUSIONS: Surgical treatment tailored to the specific pattern of dystonic activity in the individual patient is a valuable alternative in the long term management of cervical dystonia. PMID- 9408108 TI - Micturitional disturbance in patients with Guillain-Barre syndrome. AB - OBJECTIVES: To examine the frequency and pathophysiology of micturitional disturbance in patients with Guillain-Barre syndrome. METHODS: Micturitional symptoms were noted and neurological examinations made repeatedly during admission to hospital of patients with clinical and neurophysiologically definite Guillain-Barre syndrome. Urodynamic studies consisted of uroflowmetry, measurement of residual urine, urethral pressure profilometry, medium fill water cystometry, and external sphincter EMG. RESULTS: Seven of 28 (25%) patients with Guillain-Barre syndrome showed micturitional disturbance. The symptoms included voiding difficulty in six, urinary retention in three, nocturnal urinary frequency in three, and urge incontinence in two. These micturitional symptoms appeared after weakness occurred, and improved gradually along with the neurological signs. All three patients who showed retention became able to urinate. Urodynamic studies were made on four symptomatic patients two of whom underwent repeated study. Disturbed bladder sensation was noted in one patient, bladder areflexia in one, and absence of the bulbocavernosus reflex in one. Cystometry showed decreased bladder volume in two and bladder overactivity in two, one of whom had urge urinary incontinence and the other urinary retention. CONCLUSIONS: A quarter of the patients with Guillain-Barre syndrome tend to have micturitional disturbance. The patients studied had evacuation and storage disorders, as well as bladder areflexia and disturbed bladder sensation indicative of peripheral types of parasympathetic and somatic nerve dysfunction. Decreased bladder volume with bladder overactivity but no evidence of CNS involvement was also found, evidence that bladder overactivity also occurs in peripheral nerve lesions with probable pelvic nerve irritation. PMID- 9408109 TI - Random motor generation in a finger tapping task: influence of spatial contingency and of cortical and subcortical hemispheric brain lesions. AB - OBJECTIVE: To test the hypothesis that, during random motor generation, the spatial contingencies inherent to the task would induce additional preferences in normal subjects, shifting their performances farther from randomness. By contrast, perceptual or executive dysfunction could alter these task related biases in patients with brain damage. METHODS: Two groups of patients, with right and left focal brain lesions, as well as 25 right handed subjects matched for age and handedness were asked to execute a random choice motor task--namely, to generate a random series of 180 button presses from a set of 10 keys placed vertically in front of them. RESULTS: In the control group, as in the left brain lesion group, motor generation was subject to deviations from theoretical expected randomness, similar to those when numbers are generated mentally, as immediate repetitions (successive presses on the same key) are avoided. However, the distribution of button presses was also contingent on the topographic disposition of the keys: the central keys were chosen more often than those placed at extreme positions. Small distances were favoured, particularly with the left hand. These patterns were influenced by implicit strategies and task related contingencies. By contrast, right brain lesion patients with frontal involvement tended to show a more square distribution of key presses--that is, the number of key presses tended to be more equally distributed. The strategies were also altered by brain lesions: the number of immediate repetitions was more frequent when the lesion involved the right frontal areas yielding a random generation nearer to expected theoretical randomness. The frequency of adjacent key presses was increased by right anterior and left posterior cortical as well as by right subcortical lesions, but decreased by left subcortical lesions. CONCLUSIONS: Depending on the side of the lesion and the degree of cortical-subcortical involvement, the deficits take on a different aspect and direct repetions and adjacent key presses have different patterns of alterations. Motor random generation is therefore a complex task which seems to necessitate the participation of numerous cerebral structures, among which those situated in the right frontal, left posterior, and subcortical regions have a predominant role. PMID- 9408110 TI - Changes in tissue oxyhaemoglobin concentration measured using multichannel near infrared spectroscopy during internal carotid angiography. AB - OBJECTIVE: To develop an in vivo model for testing spatially resolved spectroscopy and quantified near infrared spectroscopy (NIRS) cerebral blood flow measurements. METHOD: Multiple detector NIRS has been used to study changes in tissue oxyhaemoglobin (O2Hb) concentration during selective internal carotid angiography. A significant reduction in O2Hb occurred in tissue interrogated by detectors situated between 0.7 and 4.1 cm from the NIRS light source. RESULTS: The time course of O2Hb concentration change was consistent with displacement of oxygenated blood by the radiocontrast medium from vascular beds of differing flow and NIR light attenuation. Increasing changes in O2Hb concentration per unit photon path length--predicted to occur at greater emitter-detector separations if those changes had occurred predominantly in cerebral tissue--were found in the first four seconds after injection of radiocontrast medium. However, later changes (6-10 s) were larger and were not proportional to emitter-detector separation. CONCLUSION: The findings indicate that simple assumptions regarding the distribution of the internal carotid artery blood supply to cerebral and extracerebral tissues, the photon path length through those tissues, and their relative contributions to attenuation of NIR light may not be justified. PMID- 9408111 TI - Independent segregation of von Hippel-Lindau disease and cerebral cavernomas. AB - A probable diagnosis of von Hippel-Lindau disease was made in a two generation family in which the proband had a phaeochromocytoma, renal cysts, and multiple cerebral cavernomas. His sister had multiple similar cerebral vascular lesions and his father died from renal carcinoma aged 42. Although the family did not satisfy the conventional diagnostic criteria for von Hippel-Lindau disease, an underlying germline mutation in the von Hippel-Lindau disease tumour suppressor gene was identified in the proband. Molecular genetic analysis not only confirmed the putative diagnosis of the disease in the proband but also showed that the cerebral vascular lesions segregated independently from the von Hippel-Lindau disease mutation. This report exemplifies how molecular genetic investigations can enhance the diagnosis and management of families with suspected von Hippel Lindau disease, particularly when the manifestations, as in this family, are not typical. PMID- 9408112 TI - Mononeuropathy of a distal branch of the femoral nerve in a body building champion. AB - A unique case of a body building champion with localised atrophy of the distal portion of the vastus lateralis muscle is reported; neurophysiological evaluation suggests a selective lesion of a distal branch of the vastus lateralis nerve (a motor branch of the femoral nerve). A necroscopic study in four cases was performed to better clarify the site and mechanism of nerve lesion. The data suggest that stretching and compression of the nerve has probably occurred during strenous exercise. PMID- 9408113 TI - Lymphocytic hypophysitis. PMID- 9408114 TI - Intravenous immunoglobulin treatment in lower motor neuron disease associated with highly raised anti-GM1 antibodies. AB - The effect of intravenous immunoglobulin (IVIg) treatment was studied in five patients with lower motor neuron disease associated with highly raised anti-GM1 antibodies but without evidence of conduction block on neurophysiological examination. The patients received IVIg treatment (0.4 g/kg for five consecutive days) in an open study. Only one patient responded to IVIg treatment, which was confirmed in a double blind, placebo controlled study (two placebo treatments and two IVIg treatments in a randomised order). However, after six months of maintenance IVIg treatment (0.4 g/kg weekly) muscle weakness gradually deteriorated below pretreatment levels despite continued treatment. It is concluded that the presence of raised anti-GM1 antibodies does not identify a subgroup of patients with lower motor neuron disease who respond to IVIg treatment and although some patients with lower motor neuron disease may initially respond, IVIg treatment does not seem to be sufficient as long term treatment. PMID- 9408115 TI - REM sleep motor dysfunction in multiple system atrophy: with special emphasis on sleep talk as its early clinical manifestation. AB - Various neurodegenerative diseases involving brainstem structures as one of the main pathological lesions are reported to be associated with REM sleep behaviour disorder. Full blown REM sleep behaviour disorder can be diagnosed clinically, but REM sleep motor dysfunction, a pathophysiological basis of REM sleep behaviour disorder, is difficult to detect without all night polysomnography. Twenty one consecutive patients with multiple system atrophy with no complaints of nocturnal abnormal behaviours were clinically evaluated to determine the presence of sleep related symptoms. All night polysomnography with video monitoring was performed to investigate REM sleep characteristics and patients' behaviours. In 85.7% (18 of 21) of the patients' sleep talk started or increased around or after the clinical onset of the primary diseases. REM sleep without atonia occupied more than 15% (16.2%-100%) of the REM sleep time in all but one patient. In 90.5% (19 of 21) of patients, motor events such as sleep talk and various combinations of craniofacial, orofacial, or limb movements occurred at various frequencies mostly during REM sleep without atonia. In patients with multiple system atrophy, REM sleep motor dysfunction is a common polysomnographic finding which is otherwise overlooked, and sleep talk may be its early clinical manifestation. PMID- 9408116 TI - Isolated cranial nerve palsies in multiple sclerosis. AB - During a 10 year period 24 patients with definite multiple sclerosis with isolated cranial nerve palsies were studied (third and fourth nerve: one patient each, sixth nerve: 12 patients, seventh nerve: three patients, eighth nerve: seven patients), in whom cranial nerve palsies were the presenting sign in 14 and the only clinical sign of an exacerbation in 10 patients. MRI was carried out in 20 patients and substantiated corresponding brainstem lesions in seven patients (third nerve: one patient, sixth nerve: four patients, eighth nerve: two patients). Additional abnormal findings of electro-oculography, or masseter reflex, or blink reflex, or combinations of these were found in 20 patients and interpreted in favour of a brainstem lesion at the level of the respective cranial nerve. In 11 of 14 patients with isolated cranial nerve palsies as the presenting sign of multiple sclerosis, dissemination in space was documented by MRI, and in the remaining three by evoked potentials. In patients with multiple sclerosis with isolated cranial nerve palsies, MRI is the most sensitive method of documenting dissemination in space and electrophysiological testing the most sensitive at disclosing brainstem lesions. PMID- 9408117 TI - Anosognosia for hemiplegia after a brainstem haematoma: a pathological case. PMID- 9408118 TI - Opsoclonus as a paraneoplastic manifestation of pancreatic carcinoma. PMID- 9408119 TI - Female predominance in spasmodic dysphonia. PMID- 9408120 TI - Recurrent Guillain-Barre syndrome and CNS demyelination. PMID- 9408121 TI - Non- or pseudoepileptic seizures? PMID- 9408122 TI - The World Health Organization's work on public health aspects of neurology. PMID- 9408123 TI - The likely impact of demographic changes on the incidence and prevalence of neurological disease: demography in the United Kingdom. PMID- 9408124 TI - Epidemiology of disabling neurological disease: how and why does disability occur? PMID- 9408125 TI - The economic impact of neurological illness on the health and wealth of the nation and of individuals. PMID- 9408126 TI - Primary prevention of neurological illness early in life. PMID- 9408127 TI - Prevention of acquired neurological impairment in the perinatal period. PMID- 9408128 TI - Prevention of neurological disease in later life. PMID- 9408129 TI - Neurological services and the neurological health of the population in the United Kingdom. PMID- 9408130 TI - What patients and their carers want from neurological services. PMID- 9408131 TI - Neurological care in the home. PMID- 9408132 TI - The measurement of R2, R2* and R2' in HIV-infected patients using the prime sequence as a measure of brain iron deposition. AB - Brain iron deposition was assessed at 1.5 T in the caudate nucleus, globus pallidus and frontal and parieto-occipital white matter in 28 human immunodeficiency virus (HIV)-infected patients and 15 control subjects with a new Partially Refocussed Interleaved Multi-Echo sequence by measuring 1/T2, 1/T2* and 1/T2' (i.e., R2, R2* and R2'). There were significant differences in the R2 and R2* of the caudate nucleus (p < 0.0001 and p < 0.05) and the R2, R2* and R2' of the globus pallidus (p < 0.01, p < 0.005 and p < 0.05) in HIV-infected patients compared to control subjects. There was a trend for higher values of R2, R2* and R2' in the globus pallidus and caudate nucleus in HIV-infected patients with later stage HIV disease. These results suggest that there is greater iron deposition in the basal ganglia of HIV-infected patients compared with control subjects, with a predilection for the globus pallidus. The relationship between iron deposition in the brain and various parameters of severity of HIV infection remains uncertain. PMID- 9408133 TI - Serial MR imaging of intracranial metastases after radiosurgery. AB - PURPOSE: To evaluate the spatiotemporal evolution of radiosurgical induced changes both in metastases and in normal brain tissue adjacent to the lesions by serial magnetic resonance (MR) imaging. METHODS AND MATERIALS: Thirty-five intracranial metastases of different primaries were treated in 25 patients by single high-dose radiosurgery. MR images acquired before radiosurgery were available in all patients. Sixty-three follow-up MR studies were performed in these patients including T2- and contrast-enhanced T1-weighted MR images. The average follow-up time was 9 +/- 5 months (mean +/- standard deviation [SD]). Based on contrast-enhanced T1-weighted MR images, tumor response was radiologically classified in the following four groups: stable disease was assumed if the average tumor diameter after treatment did not show a tumor shrinkage of more than 50% and an increase of more than 25%, partial remission as a shrinkage of tumor size of more than 50%, a disappearance of contrast-enhancing tumor as a complete remission, and an increase of tumor diameter of more than 25% as tumor progress. Moreover, we analysed signal changes on T2-weighted images in brain parenchyma adjacent to the enhancing metastases. RESULTS: The overall mean survival time was 10.5 +/- 7 months, with a 1-year actuarial survival rate of 40%. Stable disease, partial or complete remission of the metastatic tumor was observed in 22 patients (88%). Central or homogeneous loss of contrast enhancement appeared to be a good prognostic sign for stable disease or partial remission. This association was statistically significant (p < 0.05). Three patients (12%) suffered from tumor progression. In eight patients (32%) with stable disease or partial remission, signal changes on T2-weighted images were observed in tissue adjacent to the contrast enhancing lesions. A progression of the high signal on T2-weighted images was seen in seven of the eight patients between 3 and 6 months after therapy, followed by a signal regression 6-18 months after irradiation. CONCLUSION: MR imaging is a sensitive imaging tool to evaluate tumor response as well as the presence or absence of adjacent parenchymal changes following radiosurgery. Loss of homogeneous or central contrast enhancement on Gd enhanced MR images appeared to be a good prognostic sign for tumor response. Tumor shrinkage seems not to be dependent on time. In addition, most cases of radiation induced changes in normal brain parenchyma observed on T2-weighted images seem to be self limited. PMID- 9408134 TI - Establishing norms for age-related changes in proton T1 of human brain tissue in vivo. AB - The goal of this study was to determine the expected normal range of variation in spin-lattice relaxation time (T1) of brain tissue in vivo, as a function of age. A previously validated precise and accurate inversion recovery method was used to map T1 transversely, at the level of the basal ganglia, in a study population of 115 healthy subjects (ages 4 to 72; 57 male and 58 female). Least-squares regression analysis shows that T1 varied as a function of age in pulvinar nucleus (R2 = 56%), anterior thalamus (R2 = 51%), caudate (R2 = 50%), frontal white matter (R2 = 47%), optic radiation (R2 = 39%), putamen (R2 = 36%), genu (R2 = 22%), occipital white matter (R2 = 20%) (all p < 0.0001), and cortical gray matter (R2 = 53%) (p < 0.001). There were no significant differences in T1 between men and women. T1 declines throughout adolescence and early adulthood, to achieve a minimum value in the fourth to sixth decade of life, then T1 begins to increase. Quantitative magnetic resonance imaging provides evidence that brain tissue continues to change throughout the lifespan among healthy subjects with no neurologic deficits. Age-related changes follow a strikingly different schedule in different brain tissues; white matter tracts tend to reach a minimum T1 value, and to increase again, sooner than do gray matter tracts. Such normative data may prove useful for the early detection of brain pathology in patients. PMID- 9408135 TI - Theoretical evaluation of peripheral nerve stimulation during MRI with an implanted spinal fusion stimulator. AB - This study examines the requirements for nerve excitation near a spinal fusion implant during magnetic resonance imaging. The implant is the Spinal Fusion SpF device manufactured by Electro Biology Inc. The electric field induced within the biological medium was calculated using a three-dimensional finite difference model (described in a separate paper by Beuchler et al. from the University of Utah). Magnetic thresholds were obtained for excitation of myelinated nerve fibers that are near the implant. Minimum (rheobase) thresholds were determined for long duration dB/dt pulses, as well as strength-duration time constants (from which thresholds at other durations could be determined) for various geometries between the implant and a myelinated nerve fiber. The lowest thresholds occur when a large (20-microm diameter) fiber is situated near the bare tip of a wire from the implant, and a long duration (2 ms) stimulus is provided for which dB/dt is constant and monophasic. Magnetic thresholds for shorter durations of dB/dt are higher in accordance with a strength-duration law. In a magnetic field having a time derivative of 10 T/s that is uniform over the torso, nerve excitation is possible under worst-case conditions only for nerve fibers that are within 0.14 mm of the bare wire tip of the implant. With 20 T/s, excitation is possible only within 1 mm of the wire tip. PMID- 9408136 TI - Calculation of electric fields induced near metal implants by magnetic resonance imaging switched-gradient magnetic fields. AB - Electric (E) fields induced near metal implants by MRI switched-gradient magnetic fields are calculated by a new equivalent-circuit numerical technique. Induced E field results are found for a metallic spinal-fusion implant consisting of two thin wires connected to the metallic case of a current generator as well as for its subsections: a bare U-shaped wire, an insulated U-shaped wire, a cut insulated wire, and a generator. The presence of the metallic implants perturbs the E field significantly. Near the ends of the bare U-shaped wire, the E field is 89.7 times larger than in the absence of the wire. The greatest E field concentration occurs near the ends of the cut insulated wire, where the E field is 196.7 times greater than in the absence of the wire. In all cases, the perturbation of the induced E field by the implanted wire is highly localized within a few diameters of the wire. PMID- 9408137 TI - Image feature based automatic correction of low-frequency spatial intensity variations in MR images. AB - An automatic method of compensating for low-frequency variations in magnetic resonance images is presented. Small variations within a tissue type are modelled and a correction function is generated. The method is based completely on image features and does not need a phantom or user interaction to generate the compensation function. This image correction simplifies digital image analysis and may enhance clinical evaluation. As a result, the correction technique reduces inhomogeneity and improves contrast. Our results show that the radiofrequency response variation of coils can be reduced. The segmentation process, even with a simple threshold method, produces more reliable results when corrected images are used. The presented method is most useful for images acquired in the sagital and coronal planes with circular local coils, or using surface coils, e.g., spine coils. PMID- 9408138 TI - In vivo quantification of citrate concentration and water T2 relaxation time of the pathologic prostate gland using 1H MRS and MRI. AB - We have previously reported a striking correlation between water T2 relaxation time and citrate concentration in the normal prostate (Liney G.P.; Lowry M.; Turnbull L.W.; Manton D.J.; Knowles A.J.; Blackband S.J.; Horsman A. Proton MR T2 maps correlate with the citrate concentration in the prostate. NMR Biomed. 9:59 64; 1996). In this study we present data from similar studies of the pathologic gland. The findings support the hypothesis that measurement of both citrate concentration and water T2 relaxation time in vivo may aid the differentiation of prostatic carcinoma from benign disease and normal tissue. PMID- 9408139 TI - Radiation-induced changes in phosphorus T1 values in human melanoma xenografts studied by 31P-MRS. AB - 31P-magnetic resonance spectroscopy (MRS) has been shown to be a promising method for monitoring tumor response to radiation therapy. The purpose of the work reported here was to investigate whether the usefulness of 31P-MRS might be enhanced by measurement of spin-lattice relaxation times (T1s) in addition to resonance ratios. The work was based on the hypothesis that tumors having a high probability of being controlled locally would show shortened T1s during the treatment course due to reoxygenation and development of necrosis. BEX-t human melanoma xenografts, which show efficient reoxygenation and development of necrosis following single dose irradiation, were used as tumor models. Tumors were treated with single doses of 5.0 or 15.0 Gy and the T1s of the inorganic phosphate and nucleoside triphosphate beta resonances were measured as a function of time after irradiation by using the superfast inversion recovery method. Fractional tumor water content was determined by drying excised tumors at 50 degrees C until a constant weight was reached. The T1s in irradiated tumors were either longer than or not significantly different from those in unirradiated control tumors. The increase in the T1s following irradiation coincided in time with a radiation-induced increase in tumor water content, suggesting a causal relationship. The effects of reoxygenation and development of necrosis on T1s were probably overshadowed by the effects of tumor water content. Consequently, the usefulness of 31P-MRS in monitoring tumor response to radiation therapy might not be significantly enhanced by measurement of T1s. PMID- 9408140 TI - 1H magnetic resonance imaging and 31P magnetic resonance spectroscopy in experimental filariasis. AB - 1H Magnetic resonance imaging and 31P magnetic resonance spectroscopy (MRS) have been carried out in experimental rodent filariasis, i.e., Acanthocheilonema viteae infection in the rodent host, Mastomys coucha. The T2-weighted image of the infected host shows fine hyperintense thread like structures of adult filariid nests in the cervical region. 31P MRS of normal and infected hosts, localized over the same region of interest, show seven major peaks corresponding to phosphomonoesters (including glucose-6-phosphate, fructose-6-phosphate, fructose-1-6-diphosphate, phosphorylcholine, and adenine monophosphate or AMP), inorganic phosphate, glycerophosphorylcholine, phosphoenolpyruvate, phosphocreatine and nucleoside di- and tri-phosphates. Concentrations of phosphomonoesters (PMEs) are higher in the normal rodent compared with the infected ones. In vivo 31P MRS provides a non-invasive assessment of tissue bioenergetics and phospholipid metabolism. PMID- 9408141 TI - Dual surface coil with high-B1 homogeneity for deep organ MR imaging. AB - The theory and construction of a dual surface coil which provides good B1 homogeneity and sensitivity in a defined volume of interest is described. The probe comprises two coaxial rings, of different diameters and in different planes, which carry opposing currents of different values. Current in the second ring compensates for the roll-off of the B1 field associated with a single surface coil. Coupling between the rings and a third matching ring is by mutual inductance only. A comparison to a traditional surface coil with practical application to pig brain imaging at a field strength of 7 Tesla is shown. PMID- 9408142 TI - MR imaging of metastatic pancreatic VIPoma. AB - The MR findings in a 32-year-old man with pancreatic VIPoma and liver metastases are described. A 2-cm mass was present in the region of the tail of the pancreas that was best shown on T1-weighted fat-suppressed images as a low-signal intensity mass. Multiple liver metastases were present that showed intense peripheral ring enhancement on immediate post gadolinium spoiled gradient echo images. PMID- 9408143 TI - Allogeneic bone marrow transplantation for malignant haematological disorders. PMID- 9408144 TI - Allogeneic bone marrow transplantation for hematological malignancies- controversies and recent advances. AB - Today more than 80000 allogeneic bone marrow transplantations (BMT) have been performed worldwide. The major indications are hematological malignancies such as acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), chronic myeloid leukemia (CML) and myelodysplastic syndromes. Unrelated donors are increasingly used and there are around 4 million volunteer donors available in different registers, the largest being the National Marrow Donor Program. Molecular typing has improved the typing technique which has resulted in a decreased risk of graft versus-host disease (GVHD), lower transplant-related mortality (TRM) and improved leukemia-free survival (LFS). Using HLA-identical siblings, patients with AML in first complete remission (1 CR) and high-risk ALL in 1 CR are clear indications for BMT. However, if an HLA-identical sibling is not available, it is not known today if an unrelated bone marrow or autografting is the best option for all patients with acute leukemia in 1 CR. Because BMT is the only curable treatment for CML, a search for an unrelated donor should start as soon as it is evident that an HLA-identical sibling is not available. BMT within a year from diagnosis is of major importance for outcome. Allogeneic peripheral blood progenitor cells (PBPC) have been used as an alternative to bone marrow. Preliminary studies indicate a faster engraftment, but prospective randomized trials are necessary to establish the role of allogeneic PBPC. Umbilical cord blood has also been used as a source of allogeneic hematopoietic stem cells. Using cord blood from HLA identical siblings, engraftment seems to be delayed, but the probability of GVHD is low. Preliminary data using unrelated cord blood cells are encouraging. GVHD has an important antileukemic effect. Recently, a graft-versus-myeloma and a graft-versus-breast-cancer effect has been demonstrated. In patients who relapse after BMT, donor lymphocytes can induce remission, especially in patients with CML. With molecular techniques it is possible to detect relapse at an early stage, so called minimal residual disease. Liposomal amphotericin B has few side effects and decreased the death rate by invasive fungal infection in BMT recipients. Early diagnosis and treatment of cytomegalovirus (CMV) infection with new antiviral drugs have dramatically reduced the incidence and mortality in CMV disease. Cyclosporine combined with methotrexate is today the most widely used immunosuppressive regimen and has decreased GVHD and improved survival. However, several new immunosuppressive drugs need to be explored in clinical BMT. Immune modulation by for instance cytokines and cytokine inhibititors is a new exciting development. PMID- 9408145 TI - Tumour oxygenation assessed by polarographic needle electrodes and bioenergetic status measured by 31P magnetic resonance spectroscopy in human soft tissue tumours. AB - The purpose of this study was to evaluate the feasibility of polarographic oxygen electrode measurements and phosphorus magnetic resonance spectroscopy (31P-MRS) in extravisceral soft tissue tumours, designated to receive preoperative radiotherapy. Pretreatment tumour oxygenation was determined in 41 cases and 31P MRS was amenable to lesions in 34 patients. Biopsies were characterized histopathologically as 25 primary soft tissue sarcomas (STS), 2 recurrent STS, 9 benign and 5 other malignancies. Evaluation of phosphorus (31P) spectra was possible in 11 cases. The oxygenation status of normal tissue was higher than that of tumours, whereas no difference was found between oxygenation status of benign lesions and that of STS. There was substantial variation between tumours in the median pO2 and the bioenergetic status (beta-NTP/Pi). No correlation was found between tumour pO2 and volume (n = 25). Moreover, there was no correlation between beta-NTP/Pi and the median tumour pO2, the fraction of pO2 values < or =2.5 mmHg or tumour volume (n = 10), respectively. In conclusion, oxygen electrode assessment was found to be a clinically applicable and feasible technique for measuring tumour oxygenation status, whereas the success of 31P-MRS in human neoplasms was limited by a very poor resolution in the phosphorus signal that allowed analysis of 31P spectra in 11 tumours out of 34 cases. PMID- 9408146 TI - Weekly chemotherapy using PMitCEBO in the palliation of recurrent non-Hodgkin's lymphoma. AB - A chemotherapy schedule comprising mitozantrone, cyclophosphamide, etoposide alternating weekly with bleomycin and vincristine with oral prednisolone throughout (PMitCEBO) has been evaluated in patients relapsing after previous chemotherapy and radiotherapy as a palliative chemotherapy schedule in advanced disease. Treatment was given weekly up to a maximum of 12 weeks, the median duration being 8 weeks (range 2-12 weeks). In 20 patients, median age 70 years (range 52 82), of whom 10 had low-grade NHL and 10 relapsed intermediate/high grade NHL, the objective response rate was 75% (15/20) with a median duration of response of 12 months (range 1-27 months). Toxicity was restricted to grade 3/4 alopecia and bone marrow depression with three episodes of neutropenic sepsis but no treatment-related deaths. PMID- 9408147 TI - [18F] FDG PET in gastric non-Hodgkin's lymphoma. AB - The possibility of using [18F] FDG PET for assessment of tumor extension in primary gastric non-Hodgkin's lymphoma (NHL) was studied in 8 patients (6 high grade and 2 low-grade, one of the MALT type) and in a control group of 7 patients (5 patients with NHL without clinical signs of gastric involvement, 1 patient with NHL and benign gastric ulcer and 1 patient with adenocarcinoma of the stomach). All patients with gastric NHL and the two with benign gastric ulcer and adenocarcinoma, respectively, underwent endoscopy including multiple biopsies for histopathological diagnosis. All patients with high-grade and one of the two with low-grade NHL and the patient with adenocarcinoma displayed high gastric uptake of [18F] FDG corresponding to the pathological findings at endoscopy and/or CT. No pathological tracer uptake was seen in the patient with low-grade gastric NHL of the MALT type. In 6/8 patients with gastric NHL, [18F] FDG PET demonstrated larger tumor extension in the stomach than was found at endoscopy, and there was high tracer uptake in the stomach in two patients who were evaluated as normal on CT. [18F] FDG PET correctly excluded gastric NHL in the patient with a benign gastric ulcer and in the patients with NHL without clinical signs of gastric involvement. Although the experience is as yet limited, [18F] FDG PET affords a novel possibility for evaluation of gastric NHL and would seem valuable as a complement to endoscopy and CT in selected patients, where the technique can yield additional information decisive for the choice of therapy. PMID- 9408148 TI - Abutting orthogonal electron and photon beams in the head and neck region using asymmetrical photon beam edges. AB - Irradiation of target volumes within the ethmoid sinus and nasal cavity region has, at our institution, been performed using one anterior electron beam, abutted to two opposing lateral photon beams, in order to exclude the orbit from the irradiated volume. However, this technique give rise to larger dose hetereogeneities within the target volume, as well as undesired hot spots in the chiasma region. To reduce the inhomogenious distribution, we have introduced asymmetrical photon beam edges to broaden the penumbra of the two opposing photon beams resembling the dose gradient of the perpendicular electron beam. The achieved reduction in dose heterogeneity, measured from the differential dose volume-histogram, is in the order of 30-40% when applying the asymmetrical abutting technique compared with the conventional two opposing photon beam technique. As demonstrated by calculating the tumour control probability (TCP), the increase in dose homogeneity within the target volume, may be of clinical significance. PMID- 9408149 TI - A study of glycoconjugates in nasopharyngeal carcinoma with correlation to clinical transformation. AB - Little is known about the glycoconjugate changes in human nasopharyngeal epithelium following neoplastic changes. Glycoconjugate histochemistry (Glycine maximus (SBA), Griffonia simplicifolia II (GSA-II), Ulex europaeus (UEA-I), Arachis hypogaea (PNA) and Canavalia ensiformis (ConA)) were performed on the following nasopharyngeal biopsies: 10 adenoid tissues (benign controls), 10 chronic inflammation, 20 squamous metaplasia, 20 undifferentiated carcinoma and 5 squamous cell carcinoma. These results were correlated with the clinical transformations findings. Strong ConA and PNA staining (after neuraminidase treatment (NA)) characterized a subpopulation of squamous metaplasia subjects who later transformed to nasopharyngeal carcinoma. Strong ConA and PNA (before and after NA) depicted the majority of undifferentiated carcinoma subjects having local recurrence following irradiation therapy. In squamous metaplasia, ConA and PNA (after NA) staining may serve as a warning sign for neoplastic changes. Strong stainings for ConA and PNA (before and after NA) in undifferentiated carcinoma subjects may predict a risk for local recurrence. PMID- 9408150 TI - Extracellular matrix (ECM) and cytoskeletal modulation of cellular radiosensitivity. AB - Modulation of radiosensitivity by components of the extracellular matrix (ECM) and cytoskeletal elements has not been adequately studied. Although differences in the radiosensitivities of cells grown as monolayers, as spheroids, or grown in vitro in animal models are known, explanations have in the past neglected possible influences by the ECM and cytoskeleton. Using collagen gel cultures, it is shown that the fibrillar component of the ECM (which is responsible for cell anchorage) induces shifts in radiosensitivity. The effect is critically dependent on the affinity of the cell type towards collagen. The shifts in radiosensitivity induced by ECM alteration are manifested as changed Dq values. By applying four specific cytoskeletal poisons which either stabilize or destabilize specific cytoskeletal elements, the involvement of microfilaments and microtubuli was qualitatively appraised. Cytochalasin B, which destabilizes microfilaments (by preventing polymerization), caused a significant rise in radioresistance. This rise was due to increased D0. Although the cellular morphological change accompanying cytochalasin B treatment was essentially similar to that obtained with trypsin, the respective shifts in radioresponses were qualitatively different and opposite, suggesting differences in mechanism of action. PMID- 9408151 TI - Induction of apoptosis in neuroendocrine tumors of the digestive system during treatment with somatostatin analogs. AB - The extent of apoptosis identified by in situ DNA nick end labelling (TUNEL) on tissue samples obtained from patients with neuroendocrine tumors was correlated with the clinical outcome in patients treated with high-dose somatostatin analog (lanreotide 12 mg/day), n = 8, or other biotherapy including interferon-alpha (IFN-alpha), n = 4, low-dose somatostatin analog (octreotide or lanreotide), n = 3, or a combination of both, n = 1. Biopsies were obtained before the start of treatment and/or after 6 months and 12 months. After 6 months of treatment, 5 patients receiving high-dose somatostatin analog showed a biochemical response (decrease in different neuroendocrine tumor markers) and 4 of these showed an increase in apoptotic index (AI: percentage of apoptotic cells) by 1.94 +/- 1.71%. At 12 months, AI was also increased in patients with a biochemical response (4.22 +/- 3.93%). However, none showed a decrease in tumor size on computerized tomography (CT) and none of the patients treated with low-dose somatostatin analog or IFN-alpha showed any significant increase in AI during treatment. In an experimental model, nude mice were xenografted with the neuroendocrine cell line (BON-1). From the 2nd day of tumor implantation, they received treatment with either placebo, high-dose octreotide, IFN-alpha, or a combination of both, for 28 days. In mice receiving treatment with high-dose octreotide (300 microg/kg, t.i.d) there was a threefold increase in apoptotic cells as compared to the placebo group (p = 0.0084), while the combination group had few cells with ultra-structural changes indicating apoptosis and the IFN alpha treated group showed no significant changes. However, tumor growth inhibition was more pronounced in the combination group (p = 0.0011). This probably denotes that tumor growth inhibition could be achieved more efficiently by blocking the cell cycle than by inducing apoptosis. We concluded that treatment with high-dose somatostatin analogs may induce apoptosis in neuroendocrine tumors, while this is not found during treatment with low-dose somatostatin analogs or IFN-alpha. We also found that an increase in AI during high-dose somatostatin analog treatment was correlated with the biochemical response, but not with the tumor size as detected by CT in patients or with the tumor mass in the experimental model. PMID- 9408152 TI - VEGF and tPA co-expressed in malignant glioma. AB - Neovascularisation and migration of tumour cells are two features of highly malignant glioma. Vascular endothelial growth factor (VEGF) and tissue plasminogen activator (tPA) seem to be of importance in the process of malignancy. In the present study a topographical co-expression of tPA mRNA and VEGF mRNA (VEGF121 and VEGF165 isoforms) was demonstrated in the tumour edge of a rat malignant glioma, using in situ hybridisation. No signs of co-expression was seen in the normal brain tissue. In the normal brain the forms of VEGF mainly expressed were VEGF121, VEGF165, and VEGF189. Further studies are required to show whether VEGF and tPA are produced by the same tumour cells and to elucidate the role of this co-expression. PMID- 9408153 TI - Stage I non-Hodgkin's lymphoma treated with doxorubicin-containing chemotherapy with or without radiotherapy. AB - Seventy-six patients with stage I or IE intermediate-grade or immunoblastic non Hodgkin's lymphoma were treated with a short course of doxorubicin-containing chemotherapy with (n = 58) or without (n = 18) involved field radiotherapy. Chemotherapy consisted of 3 or 4 cycles of M-BACOD or (bleo-)CHOP. Seventy-two (97%) of the 74 evaluable patients achieved a complete response. The 3-year overall survival was 89%, recurrence-free survival 94%, and lymphoma-specific survival 93%. Patients older than 60 years also had a 3-year lymphoma-specific survival rate of as high as 92%. The International Prognostic Index was associated with overall survival (p = 0.04), but not with lymphoma-specific survival (p = 0.18). We conclude that stages I and IE intermediate-grade or immunoblastic non-Hodgkin's Hodgkin's lymphoma is highly curable if treated with short doxorubicin-containing chemotherapy and involved field radiotherapy. PMID- 9408154 TI - Treatment-related factors predisposing to chronic pain in patients with breast cancer--a multivariate approach. AB - A study was carried out to assess the factors predisposing to chronic post treatment pain in the breast area and in the ipsilateral arm in patients treated for breast cancer using two multivariate models. In the study 509 patients with non-metastasized breast cancer who were treated during 1988-1994 completed the questionnaire about pain in the operated breast and in the ipsilateral arm 10-58 months after surgery. The factors included in the analysis were: age, type of operation, size of the tumour, number of lymph nodes removed, involvement of lymph nodes, complications of surgery, intensity of the acute postoperative pain remembered by the patient, number of doses of analgesics, number of months from surgery, adjuvant radiotherapy, chemotherapy and endocrine treatment. The most important factors included in the models of chronic pain were: intensity of the acute postoperative pain, the type of operation, involvement of regional lymph nodes and radiotherapy. PMID- 9408155 TI - Cancer patterns in the Middle East--special report from the Middle East Cancer Society. AB - To update its cancer statistics, the newly established Middle East Cancer Society examined the cancer frequency patterns in Egypt and the Gaza Strip. The results revealed differing overall patterns. For men the highest frequencies were found for lymphoma, bladder cancer and cancers of the oral cavity and pharynx in Egypt, and for lung cancer, leukaemia and lymphoma in Gaza. For women, breast cancer had the highest frequency in both areas, followed by cancers of the oral cavity and pharynx in Egypt, and leukaemia and lymphoma in Gaza. The distribution of cancer occurrence by organ system also varied. In the light of the different ethnicities, lifestyles, socioeconomic levels and carcinogenic exposure among the countries of the Middle East, this kind of comparison can provide the background for more sophisticated approaches for discerning risk factors in cancer. We believe that further cooperation among participating countries will overcome the present limitations in data collection, registration and access. PMID- 9408156 TI - Acute response of pig skin to irradiation with 12C-ions or 200 kV X-rays. AB - The acute response of pig skin to treatment with high energy carbon ions (plateau region) at the Gesellschaft fur Schwerionenforschung (GSI, Darmstadt, Germany) was compared with changes after 200 kV x-irradiation. Carbon doses isoeffective to the x-ray doses were computed with a recently established model for calculation of the biological effect of heavy ions. Clinical changes and physiological symptoms (blood flow, erythema, trans-epidermal water loss, skin hydration) were scored. The parameters analyzed were maximum and mean values of each symptom during days 24 to 70 after irradiation, and the quantal endpoints for the establishment of dose effect curves were the median values of these. With exception of the maximum change in the red blood cell concentration (p < 0.02) no significant differences could be found in the response to x-rays and RBE corrected heavy ions. These results indicate that the model is valid for the calculation of biological effects of 12C-ions (plateau region) and may at least for epidermis be applied to treatment planning. PMID- 9408157 TI - Resistance pattern of 2816 isolates isolated from 17631 blood cultures and etiology of bacteremia and fungemia in a single cancer institution. AB - The resistance pattern of 2816 isolates from 17631 blood cultures and the etiology of isolates causing bacteremia and fungemia among 14591 admissions were investigated in an 80-bed single cancer institute during seven years (1990-1996) under the same empiric therapeutic antibiotic policy but with different prophylactic strategies. No change was found in the proportion of Gram-positive versus Gram-negative bacteria isolated from bacteremias (70% vs. 30%) during the past seven years. Furthermore, the proportion of coagulase-negative staphylococci and enterococci was about the same before and after the introduction of ofloxacin in prophylaxis. However, the proportion of Pseudomonas aeruginosa and Stenotrophomonas maltophilia causing bacteremia increased. There was no increase in Candida krusei and Candida glabrata after the introduction of fluconazole into our prophylactic regimen in 1992. Penicillin-resistance in viridans streptococci increased after penicillin was introduced into prophylaxis in acute leukemia in 1993. Until 1995 no quinolone-resistant Enterobacteriaceae were observed. Susceptibility to quinolones did not significantly change within the past seven years in Enterobacteriaceae after their introduction to prophylaxis in 1991, but Pseudomonas aeruginosa decreased from 90 to 58.2%. Glycopeptide resistance in enterococci and staphylococci was minimal in the observed period (0.9-4.3%). PMID- 9408158 TI - Serum erbB-2 in ductal carcinoma in situ of the breast--a marker of microinvasion? PMID- 9408159 TI - Palmar-plantar erythrodysaesthesia syndrome due to 5-fluorouracil therapy--an underestimated toxic event? PMID- 9408160 TI - Desmoplastic infantile ganglioglioma. PMID- 9408161 TI - Ion channel stability and hydrogen bonding. Molecular modelling of channels formed by synthetic alamethicin analogues. AB - Several analogues of the channel-forming peptaibol alamethicin have been demonstrated to exhibit faster switching between channel substates than does unmodified alamethicin. Molecular modelling studies are used to explore the possible molecular basis of these differences. Models of channels formed by alamethicin analogues were generated by restrained molecular dynamics in vacuo and refined by short molecular dynamics simulations with water molecules within and at either mouth of the channel. A decrease in backbone solvation was found to correlate with a decrease in open channel stability between alamethicin and an analogue in which all alpha-amino-isobutyric acid residues of alamethicin were replaced by leucine. A decrease in the extent of hydrogen-bonding at residue 7 correlates with lower open channel stabilities of analogues in which the glutamine at position 7 was replaced by smaller polar sidechains. These two observations indicate the importance of alamethicin/water H-bonds in stabilizing the open channel. PMID- 9408162 TI - Transport energetics of the Cl- pump in Aplysia gut. AB - Basolateral membranes of Aplysia foregut epithelia contain an ATP-dependent Cl- transporter (Cl- pump). Increased activity of the Cl- pump, coupled to apical and basolateral membrane depolarization, changed the Cl- transport energetics across the apical membrane but did not change the vectorially-opposite Cl- transport energetics across the basolateral membrane. PMID- 9408163 TI - Matrix-assisted laser desorption ionization mass spectrometry of membrane proteins: demonstration of a simple method to determine subunit molecular weights of hydrophobic subunits. AB - Matrix-assisted laser desorption ionization (MALDI) mass spectrometry has been used to obtain accurate molecular weight information for each subunit of several hydrophobic integral membrane proteins: cytochrome bo3 (4 subunits) and cytochrome bd (2 subunits) from E. coli, and the bc1 complex (3 subunits) and the cytochrome c oxidase (3 subunits) from Rhodobacter sphaeroides. The results demonstrate that the MALDI method is a convenient, quick, sensitive and reliable means for obtaining the molecular masses of the subunits of purified multisubunit membrane proteins. PMID- 9408164 TI - Interpretation of passive permeability measurements on lipid-bilayer vesicles. Effect of fluctuations. AB - A stochastic model for migration dynamics of solute molecules from the bulk aqueous phase to the intravesicular water pool is developed with a goal to interpret recent passive permeability measurements of bilayer membranes. Previously neglected fluctuations of the number of solubilized species in the inner water pool of a vesicle are naturally incorporated into the model. For a homogeneous one-phase bilayer, the model predicts exponential long-time asymptotics of the migration dynamics with a rate constant given by a sum of the frequencies of exit and entry of a solute molecule from/into the intravesicular water pool into/from the bulk aqueous phase. The long-time constant is directly related to the membrane permeability. PMID- 9408165 TI - The use of cis-parinaric acid to measure lipid peroxidation in cardiomyocytes during ischemia and reperfusion. AB - cis-Parinaric acid (PnAc), a fluorescent, polyunsaturated fatty acid, was used to measure lipid peroxidation during simulated ischemia and reperfusion in cultured neonatal rat cardiomyocytes. PnAc was used both as free fatty acid, inserted in the membranes following cultivation of the cells, as well as constituent of the cellular complex lipids by metabolically integrating the fatty acid during growth. In the insertion experiments a pre-incubation with DL-aminocarnitine, an inhibitor of beta-oxidation, was necessary to prevent loss of fluorescent signal. Such a pre-incubation resulted in an enrichment of PnAc in the sarcolemma: In pre treated cells 57 +/- 1.3% of total inserted PnAc is present in the sarcolemma compared to 27 +/- 5.7% in cells containing the integrated probe. Both methods to introduce PnAc into the cells were compared with respect to their sensitivity for an externally applied oxidative stress and thereafter lipid peroxidation during simulated ischemia and reperfusion was assayed. Going from normoxic to ischemic conditions lipid peroxidation did not increase and remained at a low level. When the ischemic cells were subsequently subjected to reperfusion (reintroduction of both oxygen and glucose), large scale lipid peroxidation was obvious. When, on the other hand, oxygen alone was reintroduced (reoxygenation) no increased lipid peroxidation was observed. These observations led to the conclusion that ischemia does not lead to an enhanced lipid peroxidation and that resumption of metabolic activity during reperfusion is necessary to induce lipid peroxidation. PMID- 9408166 TI - Cellular determinants of the lateral mobility of neural cell adhesion molecules. AB - The lateral mobility of the neural cell adhesion molecule (NCAM) was examined using fluorescence recovery after photobleaching (FRAP). Various isoforms of human NCAM, differing in their ectodomain, their membrane anchorage mode or in the size of their cytoplasmic domain, were expressed in NIH 3T3 cells and C2C12 muscle cells. When the various isoforms were compared in 3T3 cells, FRAP studies showed both GPI-anchored and transmembrane isoforms diffused rapidly and only small differences in either the diffusion coefficients (D) or the mobile fractions (mf) were measured, suggesting the importance of the ectodomain in regulating lateral diffusion. However, the mobility of all NCAM isoforms was greatly reduced in regions of cell-cell contact, presumably due to homophilic trans interactions between NCAMs on adjacent cells. NCAM isoforms transfected into C2C12 cells which express NCAM naturally usually displayed a significantly lower D compared to the same isoforms transfected into 3T3 cells. Thus, NCAM lateral mobility is modulated in regions where cells interact and by the structure of the host cell membrane. PMID- 9408167 TI - Effect of lithium and sodium ions on a charged membrane of dipalmitoylphosphatidylserine: a study by molecular dynamics simulation. AB - We describe a series of molecular dynamics simulations performed on a model of charged lipid bilayer (dipalmitoylphosphatidylserine) and water, in presence of sodium and lithium ions, with an atomic detail. The structure of the lipid membranes was strongly affected by the presence of lithium, as manifested by the observation of a transition from a disordered to a gel state. Concerning the mechanism of such a transition, it was associated to the dehydration that we detected in the lipid-water interface in the presence of lithium. This dehydration introduced an increase in the lipid-lipid interactions, and as a consequence, a diminution of the disorder of the membrane. When both types of ions are present in the aqueous phase, lithium shown a special affinity for the lipid membrane displacing almost all the sodium ions toward the middle of the water layer. As a result, we observed remarkable differences in the atom and electric field distributions across the lipid membrane. Concerning the diffusion and orientation of water molecules across the lipid-water interface, we also observed a strong dependency of the type ion. On the other hand, the mobility and the hydration shell of lithium and sodium ions are strongly perturbed by the presence of the charged lipid bilayer. The lipid layer was responsible for a dehydration of the ions compared to bulk water. This dehydration was compensated by an increase of coordination number of the ions with the lipid oxygens. Also, the residence times of water in the first hydration shell of lithium and sodium ions are perturbed by the presence of the lipid membrane. PMID- 9408168 TI - The proton permeability of liposomes made from mitochondrial inner membrane phospholipids: no effect of fatty acid composition. AB - The proton permeability of the mitochondrial inner membrane has been shown to correlate with the fatty acid composition of its phospholipids. In this paper, we test the hypothesis that the proton permeability of the phospholipid bilayer portion of the membrane depends on phospholipid fatty acid composition. We measured the proton permeability of liposomes made from the mitochondrial inner membrane phospholipids of eight vertebrates, representing a ten-fold range of mitochondrial proton leak and a three fold range of unsaturation index. At a membrane potential (delta psi) of 160 mV at 37 degrees C, the liposomes all had the same proton leak rate, about 30 nmol protons min-1 mg-1 phospholipid. There was no correlation between liposome proton permeability and phospholipid fatty acid composition. PMID- 9408169 TI - The role of the trehalose transporter during germination. AB - Previous studies on the resistance of yeast cells to dehydration pointed towards the protective role of trehalose and the importance of the specific trehalose transporter in guaranteeing survival. The present report demonstrates that the trehalose transporter is essential during the germination process in order to translocate trehalose from the cytosol to the external environment. Diploids that lack the trehalose transporter germinate poorly and do not form 4 spore tetrads although they accumulate trehalose and show trehalase activity. Furthermore, addition of exogenous trehalose to the germination medium enhances germination and normal segregation. The ability to transport trehalose is dominant and seems to be related to a single gene. PMID- 9408170 TI - Riboflavin transport by rabbit renal brush border membrane vesicles. AB - The present study examined riboflavin (RF) uptake by isolated rabbit renal brush border membrane (BBM). RF uptake was linear for up to 30 s and leveled off thereafter reaching an equilibrium with longer incubation. Studies on RF uptake as a function of incubation medium osmolarity indicated that the uptake was the results of transport (61.4%) into the intravesicular space as well as binding (38.6%) to membrane surfaces. The process of RF uptake was saturable as a function of substrate concentration with an apparent Km of 25.7 +/- 7.6 microM and Vmax of 75.6 +/- 14.7 pmol/mg protein/10 s. cis-Addition of unlabeled RF and its structural analogues, lumiflavin and lumichrome, inhibited the uptake of [3H]RF significantly, indicating the involvement of a carrier-mediated process in RF uptake by renal BBM. RF uptake by renal BBM was partly Na+-dependent so that when Na+ was replaced by potassium, choline, lithium or tetramethylammonium, the RF uptake was reduced to ca. 60% of the control. This Na+-dependency was unlikely to be due to Na+-cotransport mechanism because RF uptake occurred without the characteristic 'overshoot' phenomenon as for other Na+-cotransport systems and the elimination of transmembrane Na+-gradient by preloading Na+ to the intravesicular space did not affect RF uptake. In contrast, removal of Na+ eliminated the binding component of RF uptake, suggesting the requirement of Na+ for RF binding to BBM. The RF uptake was not affected when extravesicular pH was varied within the physiological pH range of 6.5 to 8.5. No effect on BBM [3H]RF uptake was found when the transmembrane electrical potential was altered by either the presence of anions with different membrane permeability (Cl- = NO3- > SO4- > gluconate-) or by using nigericin (10 microg/mg protein) with an outwardly or inwardly directed transmembrane K+ gradient. The uptake of RF by BBM vesicles was, however, inhibited by probenecid and organic anion transport inhibitors, 4,4 diiso-thiocyanatostilbene-2,2-disulfonic acid (DIDS, 1 mM) and 4-acetamido-4 isothiocyanatostilbene-2,2-disulfonic acid (SITS, 1 mM). In summary, these results demonstrate the existence of a membrane-associated, and organic anion inhibitor-sensitive, carrier system for RF uptake by renal BBM. PMID- 9408171 TI - Thylakoid membrane fluidity and thermostability during the operation of the xanthophyll cycle in higher-plant chloroplasts. AB - Barley leaves were exposed for several min to a white light of photon flux density 1000 micromol m-2 s-1, leading to a massive conversion of the xanthophyll violaxanthin to antheraxanthin and zeaxanthin in the absence of lipid peroxidation. Using electron spin resonance spectroscopy and different spin labeled stearate probes, we observed that this light treatment noticeably decreased thylakoid membrane lipid fluidity. The light-induced membrane rigidification (i) was proportional to the amount of zeaxanthin present in the membranes, (ii) was blocked by dithiothreitol, a potent inhibitor of the violaxanthin de-epoxidase, (iii) was slowly reversible in the dark, (iv) was not observed in thylakoids of an Arabidopsis mutant that has no xanthophyll cycle and (v) was accompanied by a substantial increase in the thermostability of the ionic permeability properties of the thylakoid membranes. The amount of xanthophyll cycle pigments found in photosystem II was observed to significantly decrease after illumination. Photoacoustic and chlorophyll fluorometric analyses of the illuminated leaves revealed that strong illumination decreased the quantum yield of photosynthetic oxygen evolution and the pigment antenna size of photosystem II in green light (preferentially absorbed by carotenoids) but not in red light (absorbed by chlorophylls only). Taken together in the light of previous in vitro data on carotenoids incorporated into artificial membranes, our results indicate that the xanthophyll cycle could be an 'emergency mechanism' that rapidly provides thylakoid membrane lipids with rigidifying carotenoid molecules upon sudden increase in light intensity. The significance of this mechanism for the membrane function and adaptation to stressful light and temperature conditions is discussed. PMID- 9408172 TI - Fusion activity of the influenza virus hemagglutinin does not require a transbilayer pH gradient. AB - Following reports suggesting that membrane fusion mediated by the influenza virus hemagglutinin might be dependent on a pH gradient across a putative target membrane, we have designed experiments in which this issue could be addressed directly. Accordingly, we have prepared two populations of liposomes, both simulating the plasma membrane of target cells, but with the pH of the internal aqueous medium buffered either at pH 7.4 (physiological cytosol pH) or pH 5.0 (endosomal pH at which influenza virus displays maximal fusion activity). Monitoring fusion as the relief in self-quenching of the fluorescent probe octadecylrhodamine B chloride we have found that the internal pH of the target liposomes did not influence membrane merging as mediated by the influenza virus hemagglutinin, thus demonstrating that a transmembrane pH gradient is not required for the fusion process to take place. PMID- 9408173 TI - Phase transition between hexagonal II (H[II]) and liquid-crystalline phase induced by interaction between solvents and segments of the membrane surface of dioleoylphosphatidylethanolamine. AB - We have investigated effects of several water-soluble organic solvents such as acetone, acetonitrile, and ethanol, which also have high solubility in alkane, on the structure and phase behavior of dioleoylphosphatidylethanolamine (DOPE) dispersion. X-ray diffraction data indicated that a phase transition from hexagonal II (H[II]) to liquid-crystalline (L alpha) phase in DOPE dispersion, occurred at 13% (v/v) acetone in H2O at 20 degrees C. The temperature of the L alpha-H(II) phase transition of DOPE dispersion increased with an increase in acetone concentration, and it was 37 degrees C at 20% (v/v) acetone. These results indicated that acetone stabilized L alpha phase relative to H(II) phase. Similar results were obtained in interactions of DOPE dispersions in H2O with acetonitrile or ethanol. X-ray diffraction data indicated that the H(II)-L alpha phase transition occurred at 9.0% (v/v) acetonitrile or at 9.8% (v/v) ethanol in water at 20 degrees C. The L alpha-H(II) phase transition temperature of DOPE dispersion increased with an increase in acetonitrile or ethanol concentration, and it was 66 degrees C at 20% (v/v) acetonitrile. Substitution of H2O by D2O (deuterium oxide) increased their threshold concentrations of the H(II)-L alpha phase transition induced by these organic solvents. A mechanism of these phase transitions and the effect of the substitution of H2O by D2O is proposed and discussed; an interaction free energy between solvents and the hydrophobic segments of the alkyl chains in the membrane surface, and also a packing parameter of the phospholipid may be main factors to explain these phenomena reasonably. PMID- 9408174 TI - Cutting, ageing and expression of plant membrane transporters. AB - The activity and the expression of sucrose, hexose and amino acid transporters were studied with fresh, cut or aged tissues and plasma membrane vesicles (PMV) of mature sugar beet (Beta vulgaris L.) leaves. Cutting and ageing both induced an increase of the transcripts coding for sucrose transporters and hexose transporters. No significant effect could be detected on the amino acid transporter transcripts with the probe used (aap1). A polyclonal serum directed against the Arabidopsis thaliana sucrose transporter (AtSUC1) reacted with a 42 kDa band of the sugar beet PMV, confirming previous biochemical identification of this band as a sucrose transporter. ELISA assays run with microsomal fractions and PMV using the AtSUC1 sucrose transporter probe indicated that ageing, and to a lesser extent cutting, increased the amount of sucrose transporter present in the plasma membrane. However, while cutting strongly stimulated proton-motive force driven uptake of sucrose in PMV, ageing only resulted in a slight stimulation. These data give evidence for transcriptional, post-transcriptional and post-translational controls of the activity of the sucrose transporter by mechanical treatments. Proton-motive force driven uptake of 3-O-methylglucose and valine in PMV was strongly stimulated in PMV from aged tissues, although previous data had shown that cutting did not affect theses processes. Therefore, the plant cells possess various levels of control mechanisms that allow them to regulate fluxes of the main assimilates across the plasma membrane when their natural environment is directly or indirectly altered. PMID- 9408175 TI - Assembly of the chimeric Na+/K+-ATPase and H+/K+-ATPase beta-subunit with the Na+/K+-ATPase alpha-subunit. AB - Two sets of chimeric beta-subunits were constructed from subunits of Torpedo californica Na+/K+-ATPase and pig gastric H+/K+-ATPase. Five unique restriction sites (SnaBI, EcoRV, MunI, SphI and EcoT22I) were created at equivalent positions of the respective cDNAs and were used as joining points for the construction. One set of chimeras (HxN series) was made by exchanging the 5' portion of the Na+/K+ ATPase beta-subunit cDNA with the corresponding portion of the H+/K+-ATPase beta subunit cDNA at the respective joining point. Complementary constructs were also prepared (NxH series). In the HxN series, the chimera joined at the SnaBI site formed a stable trypsin resistant complex with the Na+/K+-ATPase alpha-subunit, which was functional with respect to ATP hydrolysis and pump current generation, although the activities were less than those of the complex with the Na+/K+ ATPase beta-subunit. Trypsin resistance decreased for the complex of the chimera joined at the EcoRV site. In the NxH series, the chimeras joined at the SnaBI site and the EcoRV site formed rather trypsin-resistant complexes, but the expressions of the alpha-subunits were below 50% of the control. The chimeras joined at the MunI, SphI and EcoT22I site formed complexes susceptible to tryptic digestion. None of the chimeras in the NxH series were functional. These results suggest that at least two regions of the Na+/K+-ATPase beta-subunit [SnaBI site(Tyr40) to EcoRV site(Ile89) and EcoT22I site(Cys176) to C-terminus)] are involved in stable assembly with the Na+/K+-ATPase alpha-subunit and that the cytoplasmic domain [N-terminus to SnaBI site(Tyr40)] is functionally replaceable with the corresponding domain of the H+/K+-ATPase beta-subunit. PMID- 9408176 TI - Purification of a phosphatase which hydrolyzes phosphatidic acid, a key intermediate in glucolipid synthesis in Acholeplasma laidlawii A membranes. AB - A phosphatidic acid phosphatase (PAP; EC 3.1.3.4.), dephosphorylating phosphatidic acid (PA) to diacylglycerol (DAG), was identified and purified from the plasma membrane of Acholeplasma laidlawii A. After four purification steps, including membrane preparation, Tween 20 solubilization, preparative gel electrophoresis and electro-elution, PAP was purified about 400 times to near homogeneity. The molecular weight of PAP was according to SDS-polyacrylamide gel electrophoresis approximately 25 kDa and the enzyme was a stable and integral membrane protein. It is proposed to catalyze the first enzymatic step in the important glucolipid pathway of A. laidlawii. No essential cofactors or activator lipids were found. However, some divalent cations and phosphate analogues were potent inhibitors. Beside the in vivo substrate (PA), PAP was found to dephosphorylate p-nitrophenylphosphate. This less stringent specificity makes alternative in vivo functions for PAP plausible, the importance which is discussed. PMID- 9408178 TI - The thickness of cholesterol sulfate-containing membranes depends upon hydration. AB - The ordering of 30 mol % cholesterol (CH) or cholesterol sulfate (CS) on chain deuterated dimyristoylphosphatidylcholine (DMPC) was investigated by 2H-NMR for different hydrations. It is found that: (i) hydration has merely no influence on chain order (chain length) for DMPC-cholesterol systems, (ii) in CS-containing mixtures chain order (length) is greater at low hydration (DMPC-to-water molar ratio, Ri, of 11.3) than in excess water (Ri approximately 500) and (iii) at low hydration the ordering is about the same for CS or CH-containing systems whereas it is not at high hydration. DMPC-CS bilayer thickness is therefore very sensitive to hydration. PMID- 9408177 TI - Decreased conformational stability of the sarcoplasmic reticulum Ca-ATPase in aged skeletal muscle. AB - Sarcoplasmic reticulum (SR) membranes purified from young adult (4-6 months) and aged (26-28 months) Fischer 344 male rat skeletal muscle were compared with respect to the functional and structural properties of the Ca-ATPase and its associated lipids. While we find no age-related alterations in (1) expression levels of Ca-ATPase protein, and (2) calcium transport and ATPase activities, the Ca-ATPase isolated from aged muscle exhibits more rapid inactivation during mild (37 degrees C) heat treatment relative to that from young muscle. Saturation transfer EPR measurements of maleimide spin-labeled Ca-ATPase and parallel measurements of fatty acyl chain dynamics demonstrate that, accompanying heat inactivation, the Ca-ATPase from aged skeletal muscle more readily undergoes self association to form inactive oligomeric species without initial age-related differences in association state of the protein. Neither age nor heat inactivation results in differences in acyl chain dynamics of the bilayer including those lipids at the lipid-protein interface. Initial rates of tryptic digestion associated with the Ca-ATPase in SR isolated from aged muscle are 16(+/ 2)% higher relative to that from young muscle. indicating more solvent exposure of a portion of the cytoplasmic domain. During heat inactivation these structural differences are amplified as a result of immediate and rapid further unfolding of the Ca-ATPase isolated from aged muscle relative to the delayed unfolding of the Ca-ATPase isolated from young muscle. Thus age-related alterations in the solvent exposure of cytoplasmic peptides of the Ca-ATPase are likely to be critical to the loss of conformational and functional stability. PMID- 9408179 TI - Voltage dependent binding of annexin V, annexin VI and annexin VII-core to acidic phospholipid membranes. AB - Annexin V, VI and VII-core (delta1-107) are members of the annexin protein family and bind to acidic phospholipid membranes in a calcium dependent manner. They also show ion channel activity under certain conditions. As annexins bind peripherally to lipid membranes, ion channel formation must consist of at least two steps: An adsorption reaction regulating the binding of annexin to the membrane surface and the opening and closing of the active species controlling the channel activity. By using the baseline current through the patch clamp seal as a probe for unoccupied binding sites at the membrane, we show that the adsorption of annexins to membranes is not only calcium dependent but also strongly voltage dependent. Whereas the free transfer energies at low calcium concentrations are similar for all three annexins, the binding of annexin V becomes much tighter with higher calcium levels, compared to annexin VI and VII core. This correlates with the finding that annexin VI and VII-core display channel activity much more often than annexin V if one assumes that a high coverage of the membrane surface with annexins stabilizes the bilayer. At higher protein concentrations weaker binding is observed in agreement with the previously reported anti-cooperativity of membrane binding. PMID- 9408180 TI - Effect of a glycerol-containing hypotonic medium on erythrocyte phospholipid asymmetry and aminophospholipid transport during storage. AB - Previous studies from our laboratory have shown that under blood bank storage conditions red blood cell (RBC) ATP and lipid content were better maintained in a glycerol-containing hypotonic experimental additive solution (EAS 25) than in the conventional storage medium Adsol. The objective of this study was to determine the mechanism of the protective effect of EAS 25, by measuring transmembrane phospholipid asymmetry and the membrane integrity of stored RBCs. Split units of packed RBCs were stored in either EAS 25 or Adsol. RBCs were analyzed after 0, 42, and 84 days and vesicles shed from stored RBCs were analyzed after 84 days of storage. Phospholipid asymmetry was measured by phospholipase A2 digestion (RBCs) and activation of the prothrombinase complex (RBCs, vesicles). RBC membrane exhibited a significantly greater (P < 0.01) amount of phosphatidylethanolamine externalized after storage in Adsol than in EAS 25 (44.3% +/- 11.7 vs. 25.3% +/- 5.7, respectively). Prothrombin converting activities in RBCs were significantly lower than in shed vesicles (P < 0.001) suggesting the presence of phosphatidylserine in the outer monolayer of vesicle, but not in RBC membranes. The rates of inwardly-directed aminophospholipid transport in RBCs decreased by 50% and glutathione levels decreased by approximately 50% in both media. RBC cholesterol and phospholipid content of stored RBCs remained significantly greater (P < 0.01) in EAS 25 than in Adsol. The results indicate that despite comparable reduction in the rate of aminophospholipid transport and reduced GSH concentrations, RBC phospholipid asymmetry was better maintained during storage in EAS 25 than in Adsol. The data suggest that glycerol in the hypotonic EAS helps preserve RBC lipid organization and membrane integrity during storage. PMID- 9408181 TI - Glutathione transport system in human small intestine epithelial cells. AB - The present study characterizes for the first time a GSH specific transporter in a human intestinal epithelial cell line (I407). GSH metabolism is very important for the antioxidant and detoxifying action of intestine and for the maintenance of the luminal thiol-disulfide ratio involved in regulation mechanisms of the protein activity of epithelial cells. GSH level decreases have been related to physio-pathological alterations either of intestine or other organs. GSH specific transport systems have been identified in membranes of various cell types of rat, mice and rabbit. The presence of a Na+-independent transport system of GSH is confirmed by the similar behaviour of GSH uptake time-courses when Na+ in extracellular uptake medium was replaced with choline+ or K+ as well as by kinetic saturation and by the trans-stimulation effect on GSH uptake in GSH preloaded cells. Moreover, this transporter is activated when cations are present in extracellular medium and it is affected by membrane potential changes with an increase in GSH uptake values when membrane depolarization occurs. The present results also show a remarkable affinity and specificity of this transporter for GSH; in fact, Km value is very low (90 +/- 20 microM) and only compounds strictly related to GSH structure, such as GSH S-conjugates and GSH-ethyl ester, inhibit GSH uptake in 1407 cells. Finally, a possible hormonal control and modulation by the thiol-disulfide status of GSH transporter activity is suggested. PMID- 9408182 TI - Lateral diffusion and conductance properties of a fluorescein-labelled alamethicin in planar lipid bilayers. AB - In order to follow alamethicin diffusion within membranes under conditions of pore-formation, a fluorescein isothiocyanate (FITC) analogue was synthesized. To test the influence of the fluorescent probe addition on the pore-forming activity of the new analogue, macroscopic and single-channel experiments into planar lipid bilayers were performed. Although the apparent mean number of monomers per conducting aggregate was equivalent, the voltage-dependence of the new analogue was slightly reduced and hysteresses were broader, in agreement with the much longer duration of the open single-channels. Thus, the conducting aggregates seem to be stabilized by the introduction of the probe, presumably through the interaction of the conjugated cycles with the lipid headgroups, while the added steric hindrance may account for the slightly higher conductances of the open substates. Lateral diffusion of the labelled peptide associated with the bilayer was then investigated by the fluorescence recovery after photobleaching technique. Under applied voltage, associated with high conductance, D, the lateral diffusion coefficient, was reduced by 50% when compared to peptide at rest. These results provide new independent experimental evidence for a voltage driven insertion of the highly mobile surface-associated peptide into the bilayer as a prominent step in pore formation. PMID- 9408183 TI - Effects of dopaminergic agents on reversal of reserpine-induced impairment in conditioned avoidance response in rats. AB - Male Slc:Wistar, Std:Wistar, and Slc:F344/N rats had good acquisition of the conditioned avoidance response (CAR), while that of the male Slc:Wistar/ST, Jcl:Wistar, and Crj:Wistar rats was bad. Reserpine-induced impairment (RII) in CAR was observed 2-72 h after administration of dopaminergic (DAergic) agents in male Slc:Wistar rats. Amitriptyline (5-80 mg/kg, P.O.), imipramine, desipramine, cis-dosulepine, and trans-dosulepine at dose of 40 mg/kg, P.O. showed no antagonism against RII in CAR 20-23 h after reserpine injection (1 mg/kg, S.C.). However, the atypical antidepressive agents sibutramine (5-10 mg/kg, P.O.), bupropion (40 mg/kg, P.O.), and nomifensine (10-40 mg/kg, P.O.) exhibited antagonism against RII in CAR. The calcium channel antagonists flunarizine, nimodipine, and KP-840 at dose of 10 and 100 mg/kg, P.O., the cerebral improving agent indeloxazine (20-80 mg/kg, P.O.), the anticholinergic agent atropine (5-40 mg/kg, P.O.), 5-hydroxy-L-tryptophan (5-HTP) (40 mg/kg, I.P.), a precursor of 5 hydroxytryptamine (5-HT), and (+/-)-threo-dihydroxyphenylserine [(+/-)-threo DOPS] (20-200 mg/kg P.O.), a norepinephrine (NE) precursor, showed no antagonism against RII in CAR. The DAergic agents methamphetamine (5 mg/kg, P.O.) and amantadine (50-250 mg/kg, P.O.), L-DOPA (200 mg/kg, P.O.), and the DAergic D1/D2 receptor agonist apomorphine (0.1-1 mg/kg, S.C.) showed marked antagonism against RII in CAR. Although the DAergic D1-receptor agonist KF-38393 (0.3-30 mg/kg, I.P.) and the DAergic D2-receptor agonist quinpirole (0.3-10 mg/kg, I.P.) induced only a weak recovery of RII in CAR when they were administered alone, in contrast to a potent synergistic recovery of RII in CAR, which was observed when SKF-38393 (1 mg/kg, I.P.) and quinpirole (1 mg/kg, I.P.) were administered together. These results suggest that the DAergic nervous system rather than the adrenergic or 5 HT nervous system is involved in RII in CAR, and that both the DAergic D1- and D2 mediated nervous systems play important roles in this process. PMID- 9408184 TI - Differential effects of beta-endorphin and Met- and Leu-enkephalin on steroid hormone-induced lordosis in ovariectomized female rats. AB - The effect of intrathirdventricular (I.T.V.) injections of beta-endorphin, anti beta-endorphin antiserum, Met-enkephalin, Leu-enkephalin, and naloxone on the initial activation and final development of steroid hormone-mediated induction of female sexual receptivity was studied in ovariectomized female rats. The lordosis response to male mounts in ovariectomized rats after subcutaneous (S.C.) estradiol benzoate (EB) and progesterone (Prog) priming was facilitated by beta endorphin, and Met-enkephalin (10 microg x 5 microl(-1)), but inhibited by Leu enkephalin, when the peptides were injected into the third ventricle at the time of S.C. EB priming. A lower dose Met-enkephalin had no effects. Lordosis behavior in steroid hormone-primed rats was significantly facilitated when I.T.V. injections of Met-enkephalin were given 1 h prior to behavioral testing (47 h after EB priming). At 1 h prior to behavioral testing (47 h after EB priming), I.T.V. injection of beta-endorphin significantly inhibited lordosis behavior, especially at the higher dose of beta-endorphin (10 microg x 5 microl(-1)). Under those conditions, Leu-enkephalin had no effect. Lordosis behavior of ovariectomized female rats receiving S.C. steroid hormones and I.T.V. injection of anti-beta-endorphin antiserum was significantly inhibited when anti-beta endorphin antiserum was injected at the time of EB priming. However, lordosis was significantly facilitated when anti-beta-endorphin antiserum was injected 1 h prior to the behavior testing (47 h after EB priming). In contrast, I.T.V. injection of the opioid antagonist naloxone given either at the time of EB priming or 1 h prior to behavioral testing (47 h after EB priming) decreased lordosis behavior. The present results suggest that 1) beta-endorphin, Met enkephalin, and Leu-enkephalin have differential effects in the control of lordosis behavior; 2) the opioidergic systems may modulate initial-stage and final-stage estrogen-induced lordosis behavior; and 3) the opioidergic systems could be divided into the endorphinergic modulation-type and enkephalinergic modulation-type, based on their effects on lordosis behavior. PMID- 9408185 TI - Prenatal exposure to nicotine: effects on prepulse inhibition and central nicotinic receptors. AB - The present experiment examined effects of prenatal nicotine exposure (6 mg/kg/day via osmotic minipump) throughout gestation on prepulse inhibition of the acoustic startle response (PPI) and on the density of nicotinic acetylcholine receptors (nAchRs) in the brains of 5-week-old Sprague-Dawley rats. A total of 117 male and 103 female offspring were used. Prenatal nicotine reduced subsequent percent PPI to a 98 dB stimulus in female but not in male offspring. There was an inverse correlation between the percent of PPI and nAchR density in the cortex of male rats and the striatum of female rats. PMID- 9408186 TI - Motor dysfunction produced by tacrine administration in rats. AB - In the present study, three experiments were conducted to provide a characterization of some of the motor effects of the anticholinesterase tacrine (1.25-5.0 mg/kg I.P.) in rats. In the first experiment, tacrine was found to produce tremulous jaw movements in the dose range of 1.25-5.0 mg/kg. The second experiment examined the effects of tacrine on locomotion, and it was demonstrated that tacrine produced a dose-related suppression of open-field motor activity. In the third experiment, the effects of tacrine were assessed using operant conditioning procedures. Behavioral output during lever pressing on a fixed ratio 5 schedule was recorded by a computerized system that measured response initiation time (time from offset of one response to onset of the next) and duration for each lever press. Tacrine administration substantially depressed lever pressing response rate. This deficit was largely due to a substantial increase in the average response initiation time. Analysis of the distribution of response initiation times indicated that tacrine-treated rats made relatively few responses with fast initiation times (e.g., 0-125 ms), and also that tacrine led to a dramatic increase in the number of pauses in responding (i.e., response initiation times greater than 2.5 s). Tacrine-treated rats showed a slight increase in the average initiation time for fast responses (i.e., a slight decrease in the local rate of responding), and also showed a substantial increase in the average length of pauses greater than 2.5 s. Analysis of response durations indicated that there was an overall increase in average response duration among animals that received the higher doses of tacrine. Although tacrine-induced decreases in the local rate of responding and increases in response duration contribute to the overall deficit, the major reason why tacrine treated animals responded less was because they took much longer breaks in responding. It is possible that the tacrine-induced increases in pausing reflect a drug-induced akinesia. Thus, the present experiments indicate that tacrine impairs several aspects of motor function in the dose range tested. In view of the fact that tremor and motor slowing are classic symptoms of Parkinsonism, the present results in rats are consistent with the human literature indicating that tacrine (Cognex) can produce Parkinsonian side effects. Studies of the motor dysfunctions produced by tacrine in rats could be useful for investigating the motor side effects of tacrine in humans. PMID- 9408187 TI - Modulation of the behavioral effects of 8-OH-DPAT by estrogen and DOI. AB - The effects of female gonadal hormones on 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT)-induced flat body posture, hypothermia, and eating behavior were examined. Ovariectomized rats were injected with estradiol benzoate, estradiol benzoate, and progesterone, progesterone or vehicle on each of 2 consecutive weeks. On each week, the behavioral effects of the 5-HT1A receptor agonist, 8-OH DPAT, or the combination of both 8-OH-DPAT and 1-(2,5-dimethoxy-4-iodophenyl)-2 aminopropane hydrochloride (DOI), a 5-HT2 receptor agonist, were examined. 8-OH DPAT produced flat body posture, hypothermia, and eating behavior on each week of the experiment. Female gonadal hormones modulated eating behavior (e.g., rats treated with estradiol benzoate showed less eating after 8-OH-DPAT), but had no effect on either flat body posture or hypothermia. The 5-HT2 receptor agonist, DOI, attenuated 8-OH-DPAT's effect on flat body posture and on hypothermia, but not on eating behavior. 8-OH-DPAT's effect on all behaviors declined during the second week of the experiment. PMID- 9408188 TI - Postnatal pup brain dopamine depletion inhibits maternal behavior. AB - The interactions between dams and their pups and among siblings were investigated in litters with (a) all pups depleted of striatal dopamine by 6-hydroxydopamine (6-OHDA on PND3), (b) all pups treated with vehicle, or (c) half of the pups depleted of dopamine and the other half treated with vehicle. On PND10, two sets of four pups from each litter were videotaped in a novel environment with the dam:pup and maternal behaviors were later scored by blind observers. We observed a 70% decrease in striatal dopamine in 6-OHDA-treated pups but found no effect of treatment on pup weight gain. Dams with some or all DA-depleted pups (a) were slower to retrieve a pup and establish a nest, (b) retrieved pups less frequently, and (c) spent less time huddling with pups than dams with only vehicle-treated pups. When compared with DA-depleted pups in homogeneous litters, DA-depleted pups in mixed litters were less hyperactive and spent more time huddling with other pups than in isolation. These results suggest that DA depleted pups receive compromised maternal care but can benefit from interactions with normal siblings. PMID- 9408189 TI - Clonazepam prevents the development of sensitization to methamphetamine. AB - The GABA-benzodiazepine neurotransmission has been implicated in various forms of plasticity such as kindling and learning. The present study examined the effects of clonazepam (CZP), a GABA-benzodiazepine agonist, on the development of behavioral sensitization to methamphetamine (MA). Rats treated with MA (1 mg/kg, S.C.) for 10 days displayed significantly enhanced motor activity when tested with MA (1 mg/kg) after a 7-8-day withdrawal, indicating the development of behavioral sensitization. Pretreatment with CZP (0.5 and 2.0 mg/kg) prior to MA administration prevented the development of the phenomenon. Rats treated with CZP alone showed no difference in the motor activity compared to those treated with saline. These results suggest that stimulation of GABA-benzodiazepine receptors plays a role in the development of behavioral sensitization. PMID- 9408190 TI - Differential effects of immunologic challenge on self-stimulation from the nucleus accumbens and the substantia nigra. AB - Paralleling the effects of uncontrollable stressors, systemic administration of sheep red blood cells (SRBC) provokes brain neurotransmitter alterations, including DA variations within mesocorticolimbic regions, coinciding with or slightly preceding the peak immune response. Inasmuch as stressors disrupt responding for brain stimulation from the nucleus accumbens, possibly reflecting the anhedonic consequences of stressors, the present investigation assessed whether antigenic challenge would also influence responding for brain stimulation. Sheep red blood cell administration was found to reduce responding for brain stimulation from the nucleus accumbens, without affecting performance from the substantia nigra. The alterations of self-stimulation from the nucleus accumbens occurred at times that approximated the peak immune response. These data suggest that antigenic challenge may induce anhedonic-like effects that may be secondary to central neurochemical alterations engendered by the treatment. The possibility is also entertained that antigenic challenge may be interpreted as a stressor and contribute to alterations of affect. PMID- 9408191 TI - Anxiolytic natural and synthetic flavonoid ligands of the central benzodiazepine receptor have no effect on memory tasks in rats. AB - The naturally occurring flavonoids, chrysin (5,7-dihydroxyflavone) and apigenin (5,7,4'-trihydroxyflavone), and the synthetic compound, 6,3'-dinitroflavone have been recently reported to selectively bind with high affinity to the central benzodiazepine receptor, and to exert powerful anxiolytic and other benzodiazepine-like effects in rats. Their chemical analog, quercetin, shares none of these effects. In the present article we find that, in contrast to diazepam, chrysin, apigenin, and 6,3'-dinitroflavone have no amnestic effect on acquisition or retention of three different learning tasks (inhibitory avoidance, shuttle avoidance, and habituation to an open field), even when given at doses higher than those previously reported to be anxiolytic. Apigenin had a slight enhancing effect on training session performance and, when given posttraining, on test session retention, of crossing responses in the open field and hindered retention of inhibitory avoidance, and showed no anxiolytic action in an elevated plus maze. Unlike diazepam, none of these drugs had any analgesic effect in the tail-flick test. The data suggest that chrysin, apigenin, and 6,3' dinitroflavoine, three flavonoids derivatives possessing anxioselective effects acting on central benzodiazepine receptors, may deserve clinical trials as anxiolytic agents. PMID- 9408192 TI - Effects of microinjections of cholecystokinin and neurotensin into lateral hypothalamus and ventral mesencephalon on intracranial self-stimulation. AB - Changes in intracranial self-stimulation (ICSS) evoked from ventral tegmental area-substantia nigra (VTA-SN) and lateral hypothalamus-medial forebrain bundle (LH-MFB) before and after microinjections of sulfated cholecystokinin octapeptide (CCK-8S) and unsulfated cholecystokinin (CCK-8US), neurotensin tridecapeptide ([D Tyr11]NT(1-13) or [DTrp11]NT(1-13)) into either VTA-SN or LH-MFB were assessed. The current intensity was fixed at a level to obtain 60-70% of the maximal asymptotic rate. CCK-8S (0.10 microg/0.5 microl and 0.25 microg/0.5 microl) into VTA-SN resulted in dose-dependent decreases in VTA-SN ICSS of 38-42% and 78-92%, respectively, without affecting the ICSS of LH-MFB. Similar doses of CCK-8S injected into LH-MFB changed neither LH-MFB ICSS nor VTA-SN ICSS. CCK-8Us injected into VTA-SN or LH-MFB had no effect on ICSS in either site. Intra-VTA-SN injections of the neurotensin-1 (NT1) receptor agonist [DTyr11]NT(1-13) and the NT1 receptor antagonist [D-Trp11]NT(1-13) at doses of 5 microg/0.5 microl and 10 microg/0.5 microl decreased VTA-SN ICSS. NT1 receptor agonist and antagonist injections did not alter LH-MFB ICSS in any significant manner. Similar injections of these peptides into LH-MFB did not change the responding rates for LH-MFB ICSS or VTA-SN ICSS. Increasing the current intensity reversed the inhibitory effect of CCK-8S and [D-Trp11]NT(1-13) on VTA-SN ICSS and restored basal preinjection rates of responding. These results suggest that CCK(A) and NT1 receptor mechanisms in the ventral tegmentum in association with dopamine neurotransmission may be important in mediating the rewarding effects of VTA-SN ICSS but not LH-MFB ICSS. PMID- 9408193 TI - Chronic haloperidol administration does not block acute nicotine-induced improvements in radial-arm maze performance in the rat. AB - Nicotine has been found to improve cognitive performance in a variety of tasks including the radial maze. Nicotine has also been shown to promote the release of a variety of neurotransmitters including dopamine (DA). DA has been found to be important for nicotine's reinforcing effects. DA involvement with nicotine's cognitive effects is unclear. In the current study, the effects of acute nicotine injections (0, 0.1, 0.2, or 0.4 mg/kg) were examined on radial-arm maze performance in rats given chronic infusions the DA antagonist haloperidol (0, 0.2, or 0.6 mg/kg/day). Chronic haloperidol infusion was not found to attenuate the memory improvement caused by acute nicotine injection. In fact, the dose related nicotine-induced memory improvement was clearer in the haloperidol treated groups than in controls. This is similar to the effect of nicotine we saw in human subjects given chronic doses of haloperidol. Our previous studies demonstrated significant nicotinic-DA interactions with regard to memory function. The current results suggest that in the DA-nicotinic relationship DA stimulation is not necessary for the memory improvement caused by nicotine. PMID- 9408194 TI - The role of striatal glutamatergic system in haloperidol-induced dopamine receptor supersensitivity and effects of monosialoganglioside GM1. AB - The mechanism underlying the action of ganglioside GM1 on the increase of haloperidol-induced dopamine receptor supersensitivity was studied using the method of chemically stimulated (3H)-D-aspartate release in rat striatal slices. After a 3-week chronic haloperidol treatment the transmitter release was reduced by about 30%, with a further reduction to 40% when GM1 was applied chronically as well. This suggests that the downregulation of the glutamatergic system by chronic haloperidol treatment is potentiated by gangliosides. The acute effect of gangliosides on the stimulated (3H)-D-aspartate release from striatal slices was tested by adding GM1 to the superfusion medium. When given at a concentration of 10(-4) M, GM1 did not alter the amino acid release itself. GM1 did, however, reduce the haloperidol-enhanced (3H)-D-aspartate release to control levels and elevated the glutamate-stimulated (3H)-D-aspartate release. Binding experiments indicate that gangliosides do not directly interact with glutamate or dopamine receptors. The data are discussed in view of earlier findings that GM1 potentiates the behavioral supersensitivity following chronic haloperidol treatment without directly altering dopamine receptor supersensitivity. PMID- 9408195 TI - Effects of physostigmine on the startle in guinea pigs: two mechanisms involved. AB - The effects of the acetylcholinesterase inhibitor physostigmine (PHY) on the auditory startle reflex in guinea pigs were studied. The dose-response curve of PHY appeared bell shaped, with a maximum effect dose of 0.3 mg/kg. In addition, PHY altered the shape of the startle response. The muscarinic antagonist scopolamine (SCO) increased the startle at PHY doses above 0.3 mg/kg without affecting the PHY-induced shape of the response. The decreasing part of the startle due to PHY could be mimicked by the cholinesterase inhibitor soman in combination with 0.3 mg/kg PHY. It appeared that the decreasing part of the dose response curve at higher dose levels is caused by the cholinesterase inhibitory action of PHY and, in view of the SCO effect, is mediated by muscarinergic receptors. The increasing part of the curve is probably caused by an agonistic action of PHY on neuronal nicotinergic receptors, because the antagonist mecamylamine (20 mg/kg) antagonized the effects of 0.3 mg/kg PHY both on the deflection and shape of the startle. PMID- 9408196 TI - Effect of diazepam and A beta-carboline on open-field and T-maze behaviors in 2 day-old chicks. AB - The effects of diazepam and the beta-carboline FG 7142 in chicks were examined on several behavioral measures in open-field and T-maze tasks. In the open field, only the higher doses of diazepam affected behavior, suggesting a sedative-like effect, while FG 7142 influenced behavior as would a fear-inducing manipulation. After a low dose of either drug was injected, testing in a T-maze showed that diazepam improved and FG 7142 impaired the escape performance from the isolation chamber, without affecting the time spent in the T-corridor. In three groups of chicks selected on the basis of their first escape performance, only lower performance chicks were affected by an anxiolytic dose of diazepam. T-maze results suggest that: (a) T-maze is more sensitive than open-field test to behavior changes induced by anxiolytic doses of diazepam; (b) isolation chamber behavior could be an index of general emotionality in young chicks; (c) diazepam and FG 7142 do not modify the social motivation to escape the maze; (d) higher performance chicks present an escape behavior of a less anxious type than lower performance chicks. The results suggest that the GABAergic system is involved in the behavioral expression of fear and anxiety in young chicks. PMID- 9408197 TI - THIP, a selective gamma-aminobutyric acid receptor agonist, alters flash-evoked potentials in rats. AB - This study examined the effects of the GABA(A) agonist THIP on flash-evoked potentials (FEPs) recorded from the primary visual cortex (VC) and superior colliculus (SC) of chronically implanted hooded rats. Animals were given I.P. injections of saline, and of 8, 16, and 24 mg THIP/kg body weight on separate days. Evoked potentials were recorded at 5, 20, and 35 min following injection. Animals were tested at a standard (22.5 degrees C) room temperature. Most significant effects were observed at the 20- and 35-min recording intervals for both the 16 and 24 mg/kg doses, with effects at the 24 mg/kg dose the most pronounced. VC P1 amplitude remained unchanged, while N1 was reduced to such an extent that it became positive, ultimately blending into the rising phase of a positive component appearing between N1 and P2. This positive component had a latency of about 6 ms longer than N1, and became larger in amplitude than P1 at the 24 mg/kg dose. P2 amplitude was drastically reduced, becoming negative. In contrast, components N2 and P3 were augmented, while the amplitude of N3 was unchanged. In the SC, P1 was augmented while P3 was reduced in amplitude. A biphasic (increase/decrease) effect was observed in the N4 complex. In both the VC and SC, latencies of most components were increased, with the late components in the VC increased to the greatest extent. A mild hypothermia was observed at 16 and 24 mg/kg. The results suggest that the GABA(A) receptor plays an important role in the elaboration of the middle (N1-P2) components of FEPs recorded from the rat VC, and that GABAergic mechanisms can influence other components in the VC and SC as well. PMID- 9408198 TI - Differential behavioral responses to dopaminergic stimulation of nucleus accumbens subregions in the rat. AB - The following experiments investigated the behavioral response to local microinfusion of dopamine (DA) and selective DA agonists into the core and shell subregions of the nucleus accumbens. Rats were implanted with chronic indwelling cannulae aimed at these subregions. Two experiments were conducted. In experiment 1, the response to DA (0, 2, 5, 10 microg/0.5 microl/side), the D-1 agonist SKF 82598 (0, 0.1, 1.0 microg), the D-2/3 agonist quinpirole (0, 1, 5, 15 microg) and the D-3 preferring agonist pramipexole (0.1, 1.0, 10.0 microg) was examined in photocell activity cages. Locomotor (horizontal) and rearing (vertical) activities were measured. DA and SKF-82958 induced relatively greater increases in activity following stimulation of the shell as compared with the core. Quinpirole induced a dose-dependent suppression of activity after infusion into both sites, although the core was more sensitive to the suppressive effect than the shell. Pramipexole induced time-dependent, biphasic effects that were small in magnitude and did not differentiate between site. In experiment 2, an observation procedure was used to record behaviors (locomotion, rearing, feeding, drinking). Dopamine (0, 2, 10 microg) elicited greater increases in rearing and feeding behavior in the shell than in the core. SKF-82958 (0, 0.75 microg) enhanced locomotion and rearing to a similar extent in both subregions in this test, whereas a mixture of a low dose (0.25 microg) of the D-1 and D-2 agonists selectively induced behavioral activation in the shell. In contrast to the results in the activity cage test, quinpirole (0, 1, 5 microg) increased motor activity at the lower dose when infused into the shell but not into the core. No alterations in feeding were observed following infusion of selective agonists, and no changes in drinking were found with any of the treatments. In summary, the shell appears to be relatively more sensitive to the motor activating effects of DA agonists than the core. Moreover, circuits associated with shell may be preferentially involved in feeding. PMID- 9408199 TI - The common marmoset (Callithrix jacchus) as a model for neuroleptic-induced acute dystonia. AB - To examine whether acute dystonia is induced by neuroleptic treatment, common marmosets were treated with haloperidol orally twice a week over 25 weeks until dystonic behavior was elicited. Movement disorders such as acute dystonia were observed 6 weeks after the initial treatment, and had appeared in all treated animals by 25 weeks. Once these movement disorders were induced, they consistently reappeared after further treatment with haloperidol, and once haloperidol dosing was discontinued, the episodes vanished. Then, various neuroleptic drugs (bromperidol, chlorpromazine, risperidone thioridazine, sulpiride, tiapride, and clozapine) or a nonneuroleptic drug (diazepam) were administered orally instead of haloperidol in the above animals. All the neuroleptic drugs except for clozapine elicited similar abnormal behavior, while diazepam failed to induce any dystonia. An anticholinergic drug, trihexyphenidyl, which is known to reduce acute dystonia in patients, was also given orally to the above haloperidol-sensitized animals, followed by further treatment with haloperidol 30 min later. This clearly suppressed the induction of dystonia by haloperidol. The similarity between these findings for haloperidol-pretreated common marmosets and clinical findings suggests that the present model is useful for predicting the potential of antipsychotics to induce acute dystonia in humans. PMID- 9408200 TI - Temporal and environmental effects on quinpirole-induced biphasic locomotion in rats. AB - The dopamine D2/D3 agonist quinpirole induces suppression of locomotor activity at low doses, and suppression followed by activation at high doses when given to rats of 30 days of age and older that are immediately placed in activity monitors. The duration of suppression is longer and the level of activation is lower at 60 than at 30 days of age, suggesting that the mechanism responsible for the suppression may play a role in the lesser activation in the older rats. However, habituation limits the ability to measure the duration of locomotor suppression. Therefore, 0, 0.2, or 0.2 mg/kg quinpirole was injected S.C. either 30, 60, or 120 min before placing male or female rats of 30 or 60 days of age in activity monitors for 30 min. At both ages, both doses of quinpirole suppressed activity when the animal was placed in the monitor 30 or 60 min after injection; at 60 days the drug also suppressed activity at 120 min after injection. Previously, 0.2 mg/kg quinpirole elicited locomotor activity 60 min after injection in rats placed immediately in activity monitors at both ages. Thus, not only time after injection but novelty of the environment are critical factors in the expression of locomotor suppression or activation in response to quinpirole. PMID- 9408201 TI - Forced swim test-induced neurochemical endocrine, and immune changes in the rat. AB - The forced swim test (FST) is a behavioral paradigm that is widely used as a screening test for antidepressant activity in rodents. The objectives of the present study were to characterize the corticosterone and immune responses and in addition to examine neurotransmitter levels, in five brain regions at intervals (15, 30, 60, 90, and 120 min) following the second exposure to the FST. There was a significant but transient reduction in noradrenaline and 5-HT concentrations, in the hypothalamus 15 min post-FST exposure. 5-HT turnover in the frontal cortex and amygdala was significantly increased between 20-120 min post-FST exposure. The FST elicited a robust corticosterone response that peaked significantly at 30 min and had almost returned to baseline 120 min after exposure. There was a significant reduction in total white blood cell count 120 min after the FST, which was accompanied by a significantly reduced percentage of lymphocytes 90 and 120 min post-FST exposure. In addition, there was a significant but transient suppression of both PHA and Con A-induced lymphocyte proliferation 15 min following FST exposure. This study demonstrates that there are neurochemical changes that are coincident with the endocrine and immune changes associated with FST exposure in rats. Furthermore, this model could be used to examine the effects of manipulation of this stress response by antidepressant drugs. Such an investigation could add to our understanding of the interactions between antidepressants, stress and the neuroendocrine and immune systems. PMID- 9408202 TI - Discriminative stimulus properties of ethanol in the rat: differential effects of selective and nonselective benzodiazepine receptor agonists. AB - Rats were trained to discriminate between ethanol (1.0 g/kg; 10% v/v) and saline under a fixed ratio 10 schedule of sweetened milk reinforcement. Both diazepam [nonselective, full benzodiazepine (BZ) receptors agonist] and bretazenil (nonselective, partial BZ receptor agonist) produced dose-dependent ethanol appropriate responding (>75%). Neither diazepam nor bretazenil affected the response rate at the doses producing maximal generalisation from ethanol. In contrast, zolpidem (full BZ1 receptor agonist) and abecarnil (full BZ1/full or partial BZ2 receptor agonist) produced only moderate (<50%) ethanol-appropriate responding when tested up to doses that markedly decreased the overall response rate. These results suggest that: 1) there are no major differences between full and partial, nonselective BZ receptor agonists in their ability to substitute for 1.0 g/kg dose of ethanol; 2) stimulation of BZ1 receptors alone is not sufficient to produce ethanol-like discriminative stimulus effects in the rat. PMID- 9408203 TI - The effects of naloxone on expression and acquisition of ethanol place conditioning in rats. AB - Naloxone has been shown to facilitate extinction of ethanol-induced conditioned place preference (CPP) in mice. The present-study extended these findings by examining naloxone's effect on the expression (Experiment 1) and acquisition (Experiment 2) of place conditioning with ethanol in rats. In Experiment 1, after place conditioning with ethanol (1.8 g/kg, I.P.), groups N0, N1.5, and N10 received 0, 1.5, or 10 mg/kg naloxone before testing. As expected, ethanol produced a robust conditioned place aversion (CPA). However, naloxone had no effect on expression of CPA. In contrast to studies with mice, the endogenous opioid system does not appear to be involved in the conditioned motivational effects of ethanol in rats. In Experiment 2, groups SE1 and SE2, NS(1.5), NE(1.5), and NE(10), received ethanol alone (1.2 g/kg), naloxone alone (1.5 mg/kg), naloxone 1.5 mg/kg plus ethanol, and naloxone 10 mg/kg plus ethanol during acquisition, respectively. All naloxone-treated groups exhibited CPA. Moreover, group NE(1.5) showed a stronger CPA than group NS(1.5). The CPA produced by coadministration of naloxone and ethanol was attributed to naloxone's effects on the neural processes underlying ethanol's unconditioned aversive effects, or to other nonspecific effects on ethanol's motivational properties. PMID- 9408204 TI - Secobarbital in humans discriminating triazolam under two-response and novel response procedures. AB - Humans were trained to discriminate the benzodiazepine triazolam (0.32 mg/70 kg) from placebo under a two-response (drug vs. placebo) drug discrimination procedure. Dose-effect curves for several drugs were then determined in a crossover design using the two-response procedure and a 'novel-response procedure' that provided a novel-appropriate response for drugs unlike triazolam or placebo. Three subjects were tested with triazolam (0.1-0.32 mg/70 kg), the barbiturate secobarbital (56-177 mg/70 kg), and caffeine (320 and 560 mg/70 kg). Triazolam dose dependently increased triazolam-appropriate responding under both procedures and generally did not occasion novel-appropriate responding under the novel-response procedure. Secobarbital substituted for triazolam in the two response procedure and dose-dependently increased novel-appropriate responding as well as occasioned some triazolam-appropriate responding in the novel-response procedure. Caffeine generally occasioned placebo-appropriate responding under the two-response procedure and a mix of novel- and placebo-appropriate responding under the novel-response procedure. Triazolam and secobarbital produced qualitatively similar self-reported drug effects. These results suggest that the novel-response procedure for human drug discrimination may enhance the pharmacological selectivity of triazolam- and placebo-appropriate responding. PMID- 9408205 TI - Spontaneous epileptiform seizures but increased resistance to kindled seizures in a mutant Sprague-Dawley rat (mf/mf). AB - Approximately 30% of a breeding colony of Sprague-Dawley rats homozygous for an autosomal recessive mutation mf ("mutilated foot") associated with a peripheral sensory neuropathy have been found unexpectedly to suffer spontaneous epileptiform attacks. Seizures ranged from brief episodes of compulsive running to tonic-clonic convulsions lasting for up to 30 s, recurring at intervals of hours or days. EEG recordings during seizures showed high-voltage 8-10 Hz spike trains that abated over the ensuing 1-2 min. Interictal records were usually normal. Twice-daily kindling of the amygdala (200 microA sinewave for 1.0 s) was unexpectedly ineffective. Most of the rats that had suffered spontaneous seizures failed to develop kindled afterdischarges, even after 30 kindling stimulations. Other mf rats developed prolonged high-amplitude kindled afterdischarges that were arrested at stage 2 and failed to evolve into convulsive seizures. Hippocampal dentate granule cells of kindled mf rats, stained for zinc by Timm's method, showed significantly less mossy fibre sprouting than wild-type Sprague Dawley rats after the same number of kindled afterdischarges. A minority of the mf rats tested (2 of 14) kindled normally. Auditory stimulation (n = 23) or stroboscopic flicker (n = 14) failed to elicit seizures or running fits in any mf rat. Peripheral neuropathy corresponding to that in the mf rat, with resistance to kindling and diminished mossy fibre sprouting, have also been reported in transgenic mice with defective p75NGFR neurotrophin receptors. A homologous genetic defect in the rat could account for most of the features of the mf phenotype. PMID- 9408206 TI - Conditioned sucrose aversions produced by naloxone-precipitated withdrawal from acutely administered morphine. AB - The aversive properties of acute naloxone-precipitated morphine withdrawal were examined in the taste reactivity paradigm. Acute naloxone-precipitated withdrawal paired with sucrose solution established conditioned active rejection of the sucrose solution. Active rejection of sucrose was observed when naloxone was administered both 1 h and 22 h after morphine. When the stimulus properties of morphine were present during the conditioning trial, the conditioned sucrose aversion was only expressed when the rats were tested in the same drug state in which they had learned the aversion. However, when the stimulus properties of morphine were not present during conditioning, the aversion was expressed in the absence of the morphine state. The results suggest that palatability shifts can be conditioned to sucrose paired with acute morphine withdrawal. PMID- 9408207 TI - Cocaine impairs maternal nest building in pregnant rats. AB - The present study investigated the onset of maternal nest building in pregnant Fischer rats following chronic repeated cocaine administration. Pregnant Fischer rats were injected with saline or cocaine, 15 mg/kg, three times daily at 1-h intervals for 10 days starting on gestation day 8. Cocaine-exposed females incorporated less material into their nests and built fewer fully completed circular nests than control animals. The overall quality of the nest in cocaine exposed dams was significantly lower than that of control animals. Furthermore, cocaine exposed dams gained less weight than control females. However, no difference in number of pups, weight, or length of pups was observed between groups. Thus, it seems that cocaine disrupts the interest and skill in nest building of pregnant rats. PMID- 9408208 TI - Amphetamine-induced locomotor stereotypy in rats is reduced by a D1 but not a D2 antagonist. AB - Amphetamine produces locomotor stereotypy (repetitive routes of locomotion) in an open field. In this research we tested the ability of the D1 antagonist SKF 83566 and the D2 antagonist sulpiride to block the locomotor stereotypy produced by 2 mg/kg amphetamine. SKF 83566 decreased amphetamine-induced locomotor stereotypy; sulpiride had no consistent effect on amphetamine-induced locomotor stereotypy. There was no evidence that either antagonist potentiated the effect of the other. PMID- 9408209 TI - Adenylyl cyclase signal transduction and alcohol-induced sedation. AB - This study examined adenylyl cyclase (AC) signal transduction in alcohol sensitive brain regions of rats selectively bred for high (HAD) and low (LAD) alcohol drinking and correlated these findings with differences in sensitivity and tolerance to alcohol-induced sedation found within these lines. LAD rats were more sensitive to the sedative effects of alcohol than were HAD rats as evidenced by a shorter latency to lose the righting response (RR) after a single alcohol challenge. When time to recover the RR was compared after each of two alcohol challenges, HAD rats recovered the RR more rapidly following the second challenge compared to the first, indicating that the HAD rats rapidly developed tolerance to the sedative effects of alcohol. Tolerance did not develop in rats of the LAD line. Two months after completion of behavioral testing, adenylyl cyclase (AC) signal transduction was examined in alcohol-sensitive brain regions of rats from both lines. Immunoblot analyses indicated that LAD rats had greater Gs alpha expression in the frontal cortex (FC) and hippocampus (HIP) compared to HAD rats. Rats with the highest HIP and FC Gs alpha levels were more rapidly affected by the sedative properties of alcohol than were rats with lower Gs alpha levels. G protein expression and AC activity in the FC, HIP, cerebellum (CERE), and nucleus accumbens (ACB) were also correlated with sensitivity to the sedative properties of alcohol and with the rapid development of tolerance to this alcohol effect. The results suggest that sensitivity and tolerance to alcohol-induced sedation may be mediated in part through AC signal transduction. PMID- 9408210 TI - Rats reared in social isolation show schizophrenia-like changes in auditory gating. AB - Central sensory filtering processes can be demonstrated using a paired stimulus paradigm. Normal humans show a diminished vertex-recorded midlatency auditory evoked potential to the second of paired clicks (0.5 s apart), a phenomenon termed auditory gating. Schizophrenics routinely fail to suppress the response to the second stimulus; thus, they do not gate. Previous animal studies of auditory gating have used psychotomimetic drug administration to induce a schizophrenia like loss. However, a nonpharmacologic model of deficient gating would be advantageous. Isolation rearing of weanling rats produces impaired prepulse startle inhibition similar to that observed in schizophrenics. The present studied examined the effects of rearing status upon auditory gating. Male Sprague Dawley rats raised in social isolation (ISO) were compared to socially raised rats (SOC). Across 10 baseline recording sessions, SOC rats showed substantial gating, while ISO rats failed to gate. Abnormal auditory gating is transiently normalized by nicotine, but not haloperidol, in schizophrenics. ISO rats given nicotine bitartrate showed gating in the normal range for 60 min. By contrast, haloperidol failed to normalize gating in ISO rats. Thus, isolation rearing of weanling rats appears to produce a stable schizophrenia-like gating deficiency that shows the same pattern of response to pharmacological interventions. PMID- 9408211 TI - The effects of chlordiazepoxide on low-rate behavior are gender dependent. AB - Gender differences in anxiety have long been assumed to exist, but the experimental evidence is contradictory. It has also been suggested that antianxiety agents may have gender-dependent behavioral effects. The present experiment was designed to establish whether or not intact male and female rats behave differently when exposed to a Differential-Reinforcement of Low-Rate 72-s schedule of reinforcement (assumed to assess some of the inhibitory behavioral tendencies associated with anxiety), and whether or not the behavioral effects of acute chlordiazepoxide administration would differ between the sexes. There were no differences between male and female rats in the total number of responses, the total number of obtained reinforcers, or response efficiency in the absence of drug administration. Male and female Wistar rats were then challenged with different doses of chlordiazepoxide (vehicle, 1, 3, 10, 17, and 30 mg/kg). Low doses of chlordiazepoxide (1 and 3 mg/kg) decreased response efficiency, and medium doses (10 and 17 mg/kg) increased response efficiency in male and female rats. The highest dose of CDP (30 mg/kg) further increased response efficiency in male rats, but decreased response efficiency in female rats. These results suggest that the behavioral effects of chlordiazepoxide are dose dependent and that the effects of a large dose of chlordiazepoxide differ between male and female rats. Whether or not gender differences in drug metabolism or whether schedule contingencies played an important role in these observations remains to be determined in future experiments. PMID- 9408212 TI - Rapid recovery of self-stimulation from depression produced by the atypical neuroleptic risperidone is not prevented by 5-HT2 receptor stimulation. AB - Behavioral effects of the antipsychotic drug risperidone were tested in rats responding for variable-interval stimulation of the ventral tegmental area (VTA). Risperidone (0-0.9 mg/kg) produced a dose-dependent depression of responding in the 60 min after injection. Self-stimulation tests delayed for 30 or 120 min after injection showed that inhibition of responding by risperidone was limited in duration, with response rates recovering to pre-injection levels in a time dependent manner. Recovery occurred regardless of opportunity to engage in self stimulation, and was virtually complete at a time when receptor occupancy has been shown to be almost undiminished. The atypical properties of risperidone have been ascribed to its potent antagonist activity at 5-HT2 receptors; however, spontaneous recovery from the effects of risperidone was not prevented by simultaneous administration of a selective 5-HT2 agonist (DOI), even though DOI when given alone produced a 50-70% reduction in response rates. These results show that the inhibitory effect of risperidone on operant performance may be self limiting in a manner that is not accounted for by its pharmacokinetic properties nor by its antagonist activity at central 5-HT2 receptors. PMID- 9408213 TI - Role of 5-HT2A and 5-HT2C receptor subtypes in the two types of fear generated by the elevated T-maze. AB - To study the role of 5-HT2A and 5-HT2C receptor subtypes in anxiety, the behavioral effects of drugs that either block or stimulate these receptors were measured in an animal model of anxiety, the elevated T-maze. One arm of the maze is enclosed by walls and stands perpendicular to the two open arms. Inhibitory (passive) avoidance--representing learned fear--was measured by placing a rat at the end of the enclosed arm and recording the time to leave this arm with the four paws during three consecutive trials. After 30 s, the same animal was placed at the end of one of the open arms and the time to leave this arm with the four paws was recorded. This one-way escape response represents unconditioned fear. The I.P. injection of the preferential 5-HT2C receptor agonists mCPP and TFMPP (0.1-0.8 mg/kg), 25 min before the experimental session enhanced inhibitory avoidance. In contrast, the same drugs either tended to impair (mCPP) or significantly inhibited (TFMPP) one-way escape. The preferential 5-HT2A agonist DOI (0.03-0.3 mg/kg) did not change either inhibitory avoidance or one-way escape. Inhibitory avoidance was impaired by the selective 5-HT2C antagonists SB 200646A (3.0-30 mg/kg) and SDZ SER 082 (0.1-1.0 mg/kg), by the 5-HT2A antagonist SR 46349B (1.0-10.0 mg/kg), and by the mixed 5-HT(2A,2C) antagonist ritanserin (0.3-3.0 mg/kg). However, it was not affected by the selective 5-HT2A antagonist RP 62203 (0.25-4.0 mg/kg). All the 5-HT2 antagonists used were ineffective on one way escape. Therefore, conditioned fear seems to be tonically facilitated through 5-HT2C receptor stimulation, although the 5-HT2A receptor may also participate in its regulation. Unconditioned fear might be phasically inhibited by 5-HT2C receptor activation. PMID- 9408214 TI - Chronic administration of NMDA glycine partial agonists induces tolerance in the Porsolt swim test. AB - The Porsolt swim test (PST) was used to assess behavioral effects following acute or chronic treatment with two N-methyl-D-aspartate (NMDA) glycine partial agonists, 1-aminocyclopropanecarboxylic acid (ACPC), and D-cycloserine (DCS). Consistent with previous findings in mice, single intravenous doses of ACPC in rats produced a significant, dose-dependent reduction in immobility in the PST compared to saline. Single dose DCS also elicited significant dose-dependent reductions in PST immobility times. Single-dose ACPC or DCS (200 mg/kg) reduced immobility (p < 0.05) by 26 or 30%, respectively, compared to saline. However, multiple dosing with either ACPC or DCS (6 daily doses, 200 mg/kg) produced an apparent behavioral adaptation, as the immobility data were indistinguishable from chronic saline administration. Moreover, pretreatment with a 5-day course of ACPC or DCS promoted the development of a behavioral cross-tolerance following a sixth dose of DCS or ACPC, respectively. The development of a behavioral tolerance in the PST following chronic therapy of these drugs appears to be a general feature of glycine partial agonists. In toto, these findings support the hypothesis that chronic administration of NMDA glycine partial agonists produces a behavioral tolerance putatively through an adaptation of the NMDA receptor complex. PMID- 9408215 TI - Intranucleus accumbens amphetamine infusions enhance responding maintained by a stimulus complex paired with oral ethanol self-administration. AB - Six male Long-Evans rats were trained to self-administer 10% ethanol (v/v) during 30 min operant sessions. A licking response on an empty drinking tube resulted in the presentation of reinforcement from an automatic dipper. During the initiation of ethanol self-administration, a tone-light stimulus complex was paired with all ethanol presentations. When 10% ethanol maintained responding, guide cannulae aimed at the nucleus accumbens (NAcc) were implanted into the brain. The ability of the paired stimulus complex to reinforce a new operant response (i.e., a lever press) was then examined. To test for the development of the new response, responding on one lever resulted in presentation of only the paired tone-light stimulus complex (contingency-associated lever) while responding on an alternate lever had no programmed consequences (no contingency-associated lever). Prior to some new response sessions, amphetamine (5-20 microg/microl) was infused into the NAcc to examine the influence of dopamine on responding maintained by the stimulus complex. Ethanol intake during the sessions prior to new response testing averaged 0.49 +/- 0.07 g/g. During new response sessions no significant differences in lever pressure during no-drug conditions (control, sham, injection or vehicle injection) were observed between the contingency-associated and no contingency-associated levers. Intra-NAcc infusion of amphetamine (5-20 microg/microl) resulted in significant increases in lever pressing only on the contingency-associated lever. These data suggest that increasing NAcc dopamine levels with amphetamine enhanced the ability of the stimulus complex to function as a reinforcer. Further studies examining the ability of potentially more salient stimuli (i.e., taste of ethanol) to function as conditioned reinforcers associated with ethanol self-administration are warranted due to the apparent inability of the paired tone-light stimulus complex to function as a reinforcer without amphetamine-induced activation of the NAcc. PMID- 9408216 TI - Opioid receptor blockade during prenatal life modifies postnatal behavioral development. AB - The ontogeny of physical characteristics, spontaneous motor, and sensorimotor behaviors of preweaning rats, as well as ambulation and emotionality at weaning (day 21) were studied in rats exposed to 50 mg/kg naltrexone (NTX) or saline (controls) daily throughout gestation by maternal administration; all animals were cross-fostered to untreated mothers at birth. Morphine challenge tests and nociceptive measures revealed that this dosage of opioid antagonist blocked opioid receptors for 24 h. At birth and weaning, animals in the NTX group weighed 12 and 20%, respectively, more than control offspring. The age at which a specific physical characteristic, spontaneous motor behavior, or reflex initially appeared and the age at which 100% of the animals demonstrated a particular characteristic/behavior often were accelerated in animals prenatally exposed to NTX. The frequency of ambulation was subnormal in the NTX group, and the frequency and/or incidence of rearing, grooming, wet-dog shakes, and defecation were reduced from normal levels in these opioid antagonist-exposed rats. These results imply that interactions of endogenous opioid systems during embryogenesis are determinants of somatic, physical, and behavioral development in postnatal life. PMID- 9408217 TI - (-)-Nornicotine partially substitutes for (+)-amphetamine in a drug discrimination paradigm in rats. AB - Rats were trained in a two-lever food-reinforced operant task to discriminate (+) amphetamine (1 mg/kg) from saline. After discrimination training stabilized, test doses of (+)-amphetamine (0.0625-2.0 mg/kg), (-)-nicotine (0.1-1.0 mg/kg), or (-) nornicotine (1-10 mg/kg) were assessed for their ability to substitute for the (+)-amphetamine training dose during brief test sessions in which food reinforcement was withheld. As expected, as the test dose of (+)-amphetamine increased, there was a dose-related increase in drug-appropriate responding, with both 1 and 2 mg/kg test doses substituting fully for the (+)-amphetamine training dose. Both (-)-nicotine and (-)-nornicotine showed partial substitution (approximately 50% drug-appropriate responding) for the (+)-amphetamine training dose, with (-)-nicotine being more potent than (-)-nornicotine. Rate suppressant effects prevented the assessment of higher doses of (-)-nicotine or (-) nornicotine. Thus, while (-)-nicotine and (-)-nornicotine share similar discriminative stimulus properties, the mechanism that mediates this effect appears to differ, at least in part, from that activated by (+)-amphetamine. PMID- 9408218 TI - Effects of acute and long-term domperidone treatment on prolactin and gonadal hormone levels and sexual behavior of male and female rats. AB - Domperidone (DOMP), a dopamine D2 blocker that is unable to cross the blood-brain barrier, is an experimental tool used to induce hyperprolactinemia. Acute and long-term DOMP administration was tested in male and female rats for its effects on sexual behavior and plasma gonadal hormone levels. DOMP (4.0 mg/kg) was injected I.P. either acutely or daily for 30 days. Acute treatment failed to modify any behavioral parameter observed. The 5-day treatment stimulated and the 30-day treatment failed to inhibit sexual behavior of male rats. Serum testosterone levels were significantly reduced after 30 days of treatment in male rats. The 30-day treatment also inhibited sexual behavior and enhanced plasma progesterone levels in ovariectomized and intact female rats, respectively. The present results may be due to DOMP-induced long-term hyperprolactinemia. Alternatively, blockade of dopamine peripheral receptors induced by this treatment may also be responsible for the behavioral changes reported here. Moreover, these data suggest that female rats are more susceptible than males to the behavioral effects of long-term hyperprolactinemia. PMID- 9408219 TI - Stereotyped behavior: effects of d-amphetamine and methylphenidate in the young rat. AB - The proclivity of d-amphetamine and methylphenidate to induce perseverative motoric and vocal side effects detracts from the clinical efficacy of these stimulants in the treatment of Attention-Deficit Hyperactivity Disorder (ADHD). In an attempt to develop a model for these deleterious treatment effects, this study explored the behavioral influences exerted by d-amphetamine and methylphenidate in the young laboratory rat. This experiment revealed that doses of these stimulants that typically induce stereotypy provoke diverging behavioral profiles: while animals given 5 mg/kg d-amphetamine exhibited repetitive sniffing activity, rats treated with 30 mg/kg methyl-phenidate displayed perseverative gnawing behaviors. Although pretreatment with the serotonin synthesis inhibitor p chlorophenylalanine (PCPA) significantly attenuated both stimulant-induced stereotypies, the effect of PCPA on d-amphetamine-induced sniffing was more profound than on methylphenidate-induced gnawing. High-performance liquid chromatography (HPLC) analysis of monoamine levels in the striatum, frontal cortex, and thalamus indicated that PCPA induced an overall 89% depletion of serotonin across all conditions. These findings shed some light on the neurochemical mechanisms that underlie the differential effects of d-amphetamine and methylphenidate on stereotyped motor activity in the rat, and suggest future experiments for understanding the role of serotonin in such effects. Further, these results have implications for the differential side effects observed from each of these stimulants when used clinically in children with ADHD. PMID- 9408220 TI - Combined naloxone and fluoxetine on deprivation-induced binge eating of palatable foods in rats. AB - Opioid antagonism and serotonergic stimulation is associated with macronutrient specific hypophagia in animals. In the present study we evaluated their systemic effect alone, and in combination, at various doses, on the intake of sweet carbohydrate-rich and sweet fat-rich foods, tastes, and nutrients that are typical of binge-food items. Low-dose (1 mg/kg) naloxone, alone, preferentially suppressed fat-rich intake while low-dose (2.5 mg/kg) fluoxetine, alone, preferentially suppressed carbohydrate-rich intake. Each drug at these doses, combined with various doses of the other (2.5-10 mg/kg fluoxetine; 0.01-1 mg/kg naloxone) additively suppressed both kinds of the sweet foods. Naloxone and fluoxetine have therapeutic potential in treating binge-eating disorders. This animal study suggests what shortcomings and benefits might be expected when combining these two agents. PMID- 9408221 TI - Cathinone: an investigation of several N-alkyl and methylenedioxy-substituted analogs. AB - Structurally, methcathinone is to cathinone what methamphetamine is to amphetamine. Due to increased interest in the abuse of such agents we wished to determine if certain derivatives of cathinone would behave in a manner consistent with what is known about their amphetamine counterparts; that is, can amphetamine structure-activity relationships be extrapolated to cathinone analogs? As expected on the basis of known structure-activity relationships for amphetaminergic agents, both N-monoethylcathinone and N-mono-n-propylcathinone (N Et CAT and N-Pr CAT; ED50 = 0.77 and 2.03 mg/kg, respectively) produced amphetamine-like stimulus effects in rats trained to discriminate 1 mg/kg of (+)amphetamine from vehicle and were somewhat less potent than racemic methcathinone. In contrast, (-)N,N-dimethylcathinone or (-)Di Me CAT (ED50 = 0.44 mg/kg) was more potent than expected; although (+)N,N-dimethylamphetamine is sevenfold less potent than (+)methamphetamine, (-)Di Me CAT is only about 1.6 fold less potent than (-)methcathinone, and is essentially equipotent with ( )cathinone. In addition, although it has been previously demonstrated that 1-(3,4 methylenedioxyphenyl)-2-aminopropane (MDA) results in stimulus generalization in rats trained to discriminate (+)amphetamine or DOM from vehicle, the cathinone counterpart of MDA (i.e., MDC) resulted in partial (maximum: 58%) generalization in (+)amphetamine-trained animals, and failed to produce >7% DOM-appropriate responding in rats trained to discriminate DOM from vehicle. On the other hand, the N-methyl analog of MDC (i.e., MDMC) behaved in a manner similar to that of the N-methyl analog of MDA (i.e., MDMA); that is, a (+)amphetamine stimulus (MDMC: ED50 = 2.36 mg/kg) but not a DOM stimulus generalized to MDMC. In MDMA trained rats, stimulus generalization occured both to MDC and MDMC (ED50 = 1.64 and 1.60 mg/kg, respectively). Although this and previous studies have demonstrated that significant parallelisms exist between the structure-activity relationships of amphetamine analogs and cathinone analogs, we now report several unexpected qualitative and/or quantitative differences. It is suggested that caution be used in attempting to draw conclusions or make predictions about the activity and potency of novel cathinone analogs by analogy to the structure activity relationships derived from amphetamine-related agents; it would appear that each new cathinone analog will require individual investigation. PMID- 9408222 TI - Isradipine blocks cocaine's ability to facilitate pressing for intracranial stimulation. AB - Using rats pressing for rewarding electrical intracranial stimulation of the medial forebrain bundle, it was found that a single administration of isradipine blocked the rate-enhancing effects of cocaine (5.0 mg/kg) at doses of 3.0 and 10.0 mg/kg. Also, when isradipine (3.0 mg/kg) was administered alone (without cocaine) for 5 consecutive days, pressing for intracranial stimulation was not reduced relative to placebo levels. In another experiment, isradipine (3.0 mg/kg) persistently blocked the rate-enhancing effects of cocaine (5.0 mg/kg) across 5 consecutive days. These results support the continued investigation of isradipine as a useful adjunct to other treatments for cocaine addiction. PMID- 9408223 TI - Dietary docosahexaenoic acid increases cerebral acetylcholine levels and improves passive avoidance performance in stroke-prone spontaneously hypertensive rats. AB - We have recently shown that inferior performance in passive avoidance task is accompanied with decreased hippocampal choline (Ch) in stroke-prone spontaneously hypertensive rats (SHRSP) compared with normotensive control Wistar-Kyoto rats (WKY). We also reported that dietary docosahexaenoic acid (DHA) suppresses the development of hypertension and stroke-related behavioral changes, resulting in the prolongation of the life span of SHRSP. In this study, we examined the effect of dietary DHA on the cerebral acetylcholine (ACh) levels and learning performance in passive avoidance tasks in SHRSP. The arachidonic acid decreased and the DHA increased in plasma lipids dose dependently with dietary DHA treatments, which decreased the systolic blood pressure in SHRSP. Dietary DHA significantly restored the significantly inferior learning performance in passive avoidance response observed in control SHRSP (DHA 0%). Furthermore, the hippocampal ACh levels were correlated positively with the total response latency in passive avoidance tasks. These results suggest that cholinergic dysfunction in the brain of control SHRSP is responsible, at least in part, for the impaired learning ability and the dietary DHA ameliorates this performance failure. PMID- 9408224 TI - Influence of prior experience on mice behavior using the four-plate test. AB - A single prior undrugged exposure to the four-plate test reduces punished responding on retest at intervals ranging from 24 h to 42 days. Furthermore, prior experience attenuates the anxiolytic response to the benzodiazepines diazepam (0.25 to 2 mg/kg) and lorazepam (0.5 to 4 mg/kg). The result was first discussed in term of "one trial tolerance." The anxiety baseline was increased during the retest, which counteracted the anxiolytic action of benzodiazepines. To ascertain if memory processes are also implicated, the cholinergic drugs scopolamine and oxotremorine were used. Additional experiments with the GABAergic inverse agonist FG7142 and with the 5-HT1A receptor agonist 8-OH-DPAT were also performed. Administration of scopolamine and 8-OH-DPAT-induced weak impairment of memory, when administered before the second trial, but no effect was seen with cognition enhancing agents. PMID- 9408226 TI - Effects of varying marijuana potency on deposition of tar and delta9-THC in the lung during smoking. AB - To determine whether smoking more, compared to less, potent marijuana (MJ) cigarettes to a desired level of intoxication ("high") reduces pulmonary exposure to noxious smoke components, in 10 habitual smokers of MJ, we measured respiratory delivery and deposition of tar and delta9-tetrahydrocannabinol (THC), carboxyhemoglobin (COHb) boost, smoking topography, including cumulative puff volume (CPV) and breathholding time, change in heart rate (deltaHR) and "high" during ad lib smoking of 0, 1.77, and 3.95% MJ cigarettes on 3 separate days. At each session, subjects had access to only a single MJ cigarette. On average, smoking topography and COHb boost did not differ across the different strengths of MJ, while THC delivery, as well as HR, were significantly greater (p < 0.01) and tar deposition significantly less (p < 0.03) for 3.95% than 1.77% MJ. Although individual adaptations in smoking topography for 3.95% compared to 1.77% MJ were highly variable, three subjects with the lowest 3.95% MJ:1.77% MJ ratios for CPV also displayed the lowest 3.95% MJ:1.77% MJ ratios for tar deposition. In vitro studies using a standardized smoking technique revealed a mean 25% lower tar yield from 3.95% than 1.77% MJ (p < 0.05), but no difference between 1.77% and 0% marijuana. Under the conditions of this study, we conclude that tar delivery is reduced relative to THC content in a minority of subjects, and this reduction appears to be due to a reduced intake of smoke (decreased CPV) and/or a reduced tar yield from the stronger MJ preparation. PMID- 9408225 TI - Evaluation of cannabimimetic discriminative stimulus effects of anandamide and methylated fluoroanandamide in rhesus monkeys. AB - In previous research arachidonylethanolamide (anandamide) has been shown to produce behavioral effects in mice characteristic of psychoactive cannabinoids, including antinociception, catalepsy, hypothermia, and hypomotility. However, differences have also been found between anandamide and delta9 tetrahydrocannabinol (delta9-THC), with anandamide having lower potency, a more rapid onset, and shorter duration of action than delta9-THC. Although it can produce delta9-THC like discriminative stimulus effects in rats, anandamide also produces concomitant response rate decreasing effects, whereas with delta9-THC there is a better separation of these two behavioral effects. The present study was designed to examine the discriminative stimulus effects of anandamide in rhesus monkeys trained to discriminate delta9-THC from vehicle. While anandamide failed to produce reliable substitution for delta9-THC and did not reduce response rates at doses up to 10 mg/kg, 2-methylarachidonyl-2'-fluoroethylamide (methylated fluoroanandamide), a putative stable analog of anandamide, produced full dose-dependent substitution for delta9-THC at doses that caused no significant changes in response rates. These results suggest that systemically administered anandamide may be metabolized in monkeys before behaviorally active concentrations could reach the brain and further suggest that the metabolically more stable analog of anandamide, methylated fluoroanandamide, may aid in the discovery of functional properties of the endogenous cannabinoid system. PMID- 9408227 TI - Effects of dorsal noradrenergic bundle lesions on recovery after sensorimotor cortex injury. AB - Several lines of evidence suggest that the recovery of the ability of rats to traverse a narrow beam after unilateral injury to the sensorimotor cortex is noradrenergically mediated. We tested the hypotheses that the influence of norepinephrine on beam-walking recovery occurs, at least partially, through effects in the contralateral and/or ipsilateral cerebral cortex. Rats had either a selective left or right 6-hydroxydopamine lesion or sham lesion of the dorsal noradrenergic bundle (DNB) 2 weeks before suction-ablation or sham injury of the right sensorimotor cortex. The rats' abilities to perform the beam-walking task were measured over the 10 days following cortex surgery. DNB lesions did not affect the initial severity of the beam-walking deficit and had no effect on the performance of the task in rats with sham cortex injuries. Lesions of the contralateral but not ipsilateral DNB significantly impaired recovery. Further, in cortically lesioned rats with contralateral DNB lesions, norepinephrine content in the cerebral cortex opposite to the sensorimotor cortex lesion was significantly correlated with recovery. These data suggest that the effect of norepinephrine on recovery of beam-walking ability may be partially exerted in the cerebral cortex contralateral to the injury. PMID- 9408228 TI - Interactions of ethanol with nicotine, dizocilpine, CGP 40116, and 1-(m chlorophenyl)-biguanide in rats. AB - The present study examined the effect of ethanol (0.25-1.0 g/kg, I.P.) alone and in combination with drugs affecting different ligand-gated ion channels on a horizontal locomotor activity of male Wistar rats. None of the drugs given alone affected the locomotor activity. Similarly, combining ethanol either with nicotine (0.1 or 0.6 mg/kg, S.C.) or the competitive NMDA receptor antagonist, CGP 40116 (0.5 mg/kg, I.P.) did not result in any significant changes in ambulation. On the other hand, a significant hyperadditive interaction between ethanol (0.5 or 1.0 g/kg) and the uncompetitive NMDA receptor antagonist, dizocilpine (0.1 mg/kg, I.P.) was found. Thus, a combined administration of ethanol and dizocilpine produced a marked stimulation of the locomotor activity. Combining 1.0 g/kg ethanol with the 5-HT3 receptor agonist, 1-(m-chlorophenyl) biguanide (5.0 mg/kg, I.P.) tended to produce locomotor stimulation. Our results suggest the existence of interaction between ethanol and the NMDA receptor complex in mediation of locomotor stimulation. Alternatively, a common neurotransmitter system (other than glutamatergic) mediate central stimulatory effects of ethanol and dizocilpine. A possible role of dopamine in this interaction is being discussed. PMID- 9408229 TI - Psychological stress does not affect plasma catecholamines in subjects with cardiovascular disorder. AB - Whereas the effects of cardiac transplantation on the catecholamine response to physical exercise have been studied previously, the impact on psychological stress is unknown. Here, the arterial catecholamine response to the Stroop test of patients with an orthotopic heart transplant (OHT) was compared with that in subjects who had received a coronary artery bypass graft (CABG) or who were in heart failure and destined for a heart transplant (HF). Subjects were tested whilst sitting and their usual drug therapy was maintained. The Stroop test increased subjects' subjective tension but did not affect arterial concentrations of adrenaline or noradrenaline in any group of subjects. Also, the concentration of both catecholamines was significantly higher in OHT and CABG subjects than in the HF group, but their relative concentration was unaffected by cardiovascular status or stress. It is concluded that the absolute concentrations of arterial catecholamines, but not their relative concentrations, depend on clinical status. Moreover, under these test conditions, subjects with a history of cardiovascular disorder do not show the normal catecholamine response to psychological stress. PMID- 9408230 TI - Acute testicular ischemia results in germ cell-specific apoptosis in the rat. AB - Testis torsion-induced aspermatogenesis is not necessarily due to permanent loss of blood flow nor to dysfunctional Leydig cells or Sertoli cells. This investigation was undertaken to gain further insight into the mechanism underlying torsion-induced germ cell loss. Male rats were subjected to 1-h or 2-h ischemia-inducing torsion, and testes were examined at either 1, 2, 4, 24, or 48 h after torsion, depending on the study. Testes were examined for evidence of 1) in situ apoptosis by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate (dUTP)-biotin nick-end labeling (TUNEL) technique, 2) apoptosis by the DNA "laddering" technique, 3) leukocyte margination and diapedesis in testicular vessels by immunocytochemical and histological techniques, and 4) testicular lipid peroxidation by the thiobarbituric acid reactive substances assay. The first TUNEL evidence for torsion-induced apoptosis was at 4 h after repair of 1-h torsion. Induction of apoptosis was confirmed by the electrophoretic laddering of DNA fragments. It was hypothesized that apoptosis was induced by reactive oxygen species arising from reperfusing leukocytes. A significant increase in both leukocyte margination and diapedesis occurred 4 h after torsion repair as did a significant increase in intratesticular lipid peroxidation products. These events were contemporaneous with the first appearance of apoptosis and consistent with the hypothesis that post-torsion, germ cell-specific apoptosis is induced by reactive oxygen species. PMID- 9408231 TI - On the synthesis and secretion of rat seminiferous tubule proteins in vivo after ischemia and germ cell loss. AB - This study was undertaken to determine whether alterations in Sertoli cell protein synthesis and secretion were important precursors to germ cell loss after ischemic insult to the testis. Ischemia was induced by a 1-h, 720 degrees spermatic cord torsion, and this was shown to cause a loss of germ cells over a 15-day period. Seminiferous tubules were perifused in vivo with [35S]methionine. Lumen fluid (LF) was collected by in vivo micropuncture, and seminiferous tubule extract (TE) was collected after tubule homogenization and centrifugation. Electrophoresis of proteins in these fluids followed by autoradiography of radiolabeled proteins allowed examination of synthesized, i.e., TE, and secreted, i.e., LF proteins. No consistent changes were detected in synthesized or secreted proteins prior to the major loss of germ cells; thus, major changes in the capacity of Sertoli cells for protein assembly and transport are not a preliminary feature of post-ischemia germ cell loss. Changes in specific protein synthesis and secretion were also modest in this in vivo environment after germ cell loss. Overall protein synthesis appeared reduced as loss of germ cells progressed, but one protein whose amino acid sequence confirmed identity with a testis-specific stress protein (hst70) was up-regulated after ischemia and germ cell loss. PMID- 9408232 TI - Transcriptional regulation of human placental corticotropin-releasing factor by prostaglandins and estradiol. AB - The mechanism of labor initiation in humans has not been completely elucidated. Prostaglandins, estrogens, and corticotropin-releasing factor (CRF) have all been shown to affect uterine myocytes and enhance uterine contractility. There are also indications that these uterine regulators have additional effects on other sites involved in labor and that they may act in concert or, perhaps, by regulating each other. Therefore, we evaluated the CRF promoter for transcriptional regulation by prostaglandins and estrogens. Human placental choriocarcinoma cell lines were transfected with CRF-luciferase reporter genes and treated with prostaglandins. Prostaglandin E2 (PGE2), but not prostaglandin F2alpha (PGF2alpha), stimulated CRF-luciferase expression in choriocarcinoma cell lines via a cAMP-dependent pathway. A combination of transfections and in vitro binding studies tested for potential regulation of CRF by estrogen receptor (ER). ER neither regulated the CRF promoter nor interacted with steroid response half sites from the CRF promoter. Our results provide a molecular regulatory link between PGE2 and CRF, two compounds that enhance uterine contractile function. Combined with the stimulation of prostaglandin release by CRF, these data support a potentially important "feed-forward" regulatory loop involving CRF and PGE2 in parturition. In contrast, we found no evidence for direct effects of estrogens or PGF2alpha on CRF transcription. PMID- 9408233 TI - Osteopontin is the 55-kilodalton fertility-associated protein in Holstein bull seminal plasma. AB - Previously we identified four proteins in seminal plasma that were associated with bull fertility. The purpose of this study was to identify the 55-kDa protein prevalent in seminal plasma of higher-fertility males. The 55-kDa protein was quantified by video densitometry in two-dimensional electrophoresis gels of seminal plasma from 26 bulls of known fertility. Relative density of the 55-kDa protein was positively correlated (r = 0.48) with bull fertility. The 55-kDa (pI 4.5) fertility-associated protein spot was isolated by electroelution after two dimensional PAGE separation of seminal plasma of 36 bulls. N-terminal sequence analysis of the pure protein yielded a 15-amino acid sequence (VKPXSSGXSEEKQLN) that was 86% homologous to bovine osteopontin-k precursor. Polyclonal antiserum generated against the 55-kDa protein reacted with a single spot in two dimensional PAGE Western blots of seminal plasma. Western blot analyses using polyclonal antisera generated against the amino terminus (LF123) and carboxyl terminus (LF124) of human recombinant osteopontin confirmed that the 55-kDa polypeptide was osteopontin. Partially purified 55-kDa protein was obtained by HPLC-MonoQ column chromatography. Protein characterization revealed that the 55 kDa protein was glycosylated, but not phosphorylated, consistent with the identity of the 55-kDa protein as osteopontin. PMID- 9408234 TI - Signaling pathways controlling trophoblast cell differentiation: Src family protein tyrosine kinases in the rat. AB - Trophoblast giant cell differentiation is characterized by endoreduplication and expression of members of the prolactin (PRL) gene family and can be simulated in vitro via manipulations of the Rcho-1 trophoblast cell line. The regulation of trophoblast cell proliferation and differentiation involves tyrosine protein kinase signaling pathways. Treatment of Rcho-1 trophoblast cells with tyrosine kinase inhibitors disrupted differentiation-dependent expression of members of the PRL gene family and cytoskeletal organization. Activated p60c-src, p62c-yes, and p53/56lyn were present in the Rcho-1 rat trophoblast cell line and in differentiated trophoblast cells isolated from the developing rat placenta. p60c src and p62c-yes were active in proliferating and differentiating trophoblast cells. During proliferation, p62c-yes exhibited distinct associations with other phosphoproteins (34, 66, 76, and 150 kDa). p53/56lyn was activated only in differentiating trophoblast cells. p53/56lyn showed a differentiation-dependent accumulation in cytoskeletal and membrane fractions, whereas p60c-src levels were virtually invariant in both fractions. Expression patterns of csk, a negative regulator of Src family kinase activities, were not consistent with its involvement in the differentiation-dependent activation of p53/56lyn; however, there was some indication of the participation of a tyrosine phosphatase in the regulation of p53/56lyn. In conclusion, p60c-src, p62c-yes, and p53/56lyn patterns of activation in trophoblast cells are consistent with their involvement in the control of trophoblast cell proliferation and differentiation. PMID- 9408235 TI - Expression of mitotic cyclin B1 is not confined to proliferating cells in the rat testis. AB - Spermatogenesis is a precisely controlled and timed process comprising mitotic divisions of spermatogonia, meiotic divisions of spermatocytes, and the maturation and differentiation of haploid spermatids. Cell proliferation is controlled by genes involved in the regulation of the cell cycle. Among the principal regulatory proteins are cyclins, which are categorized according to their appearance during the cell cycle. B-type cyclins are mitotic cyclins and function at the G2/M transition of the cell cycle. We have investigated the expression and regulation of cyclin B1 during rat spermatogenesis. Rat cyclin B1 was isolated from a testis cDNA library and further used as a probe to detect mRNA expression. Northern blot hybridization of testis mRNA revealed the presence of a single 1.7-kilobase transcript. In situ hybridization showed stage-specific expression during spermatogenesis with highest expression found in late pachytene spermatocytes and early round spermatids. This pattern was confirmed in fractions of isolated germ cells. Immunocytochemistry displayed highest protein levels in round spermatids. Depletion of gonadotropins did not change the quantitative and qualitative expression pattern of cyclin B1. Therefore, the signals triggering the onset of cyclin B1 expression seem not to originate from the pituitary gonadal endocrine axis and might therefore be paracrine factors originating within the germinal epithelium. Our observations suggest that cyclin B1 plays a hitherto unknown role in spermatid maturation in addition to its known function in dividing cells. PMID- 9408236 TI - Role of diameter differences among follicles in selection of a future dominant follicle in mares. AB - Follicles > or = 5 mm were ablated in pony mares by a transvaginal ultrasound guided technique on Day 10 (ovulation = Day 0). Follicle emergence (at 15 mm, experiment 1; at 6 mm, experiment 2) and development of the new wave was monitored by transrectal ultrasound. Deviation was defined as the beginning of a marked difference in growth rates between the two largest follicles. In experiment 1, mares were grouped (n = 4 per group) into controls, ablation controls (ablations at Day 10 only), and a two-follicle model (periodic ablation sessions so that only the two largest follicles developed). There were no significant indications that the two-follicle model altered follicle diameters, growth rates, or time intervals of the two retained follicles at or between events (follicle emergence, deviation, and ovulation). In experiment 2, the two follicle model (n = 14) was used for follicle and hormonal characterization and hypothesis testing, without the tedious and error-prone necessity for tracking many (e.g., 20) individual follicles. The future dominant follicle emerged a mean of 1 day earlier (p < 0.008) than the future subordinate follicle, the growth rates for the two follicles between emergence and deviation (6 days later) did not differ, and the dominant follicle was larger at the beginning of deviation (23.1 +/- 0.8 mm versus 19.6 +/- 0.9 mm; p < 0.0001). Mean FSH and LH concentrations increased (p < 0.05) concomitantly from emergence of the future dominant follicle and peaked 3 days later when the follicle was a mean of 13 mm. Thereafter, the two hormones disassociated until ovulation: FSH decreased and LH increased. Results supported the hypothesis that the future dominant follicle has an early size advantage over future subordinate follicles and indicated that the advantage was present as early as 6 days before deviation. PMID- 9408237 TI - Decline in serum follicle-stimulating hormone concentrations alters key intrafollicular growth factors involved in selection of the dominant follicle in heifers. AB - Declining FSH after a transient rise coincides with selection of a dominant follicle (DF) and atresia of the remaining cohort follicles (subordinates) in cattle. The objectives of this study were to determine 1) whether intrafollicular amounts of inhibins, activin-A, insulin-like growth factor I (IGF-I), and IGF-I binding proteins (IGFBP) are altered during selection of the first-wave dominant follicle (DF1) and 2) whether these biochemical markers are FSH dependent. Beef heifers received six or eight 6-h injections of saline (controls) or eight 6-h injections of recombinant bovine FSH (1 mg/injection) at 38 to 42 h after estrus (Day 0). Daily ultrasound scanning was used to define selection of DF1. Controls (n = 6 per group) were ovariectomized 1) on Day 3 of the estrous cycle before DF1 selection (preselection follicles) and 2) after DF1 selection on Day 4.8 +/- 0.5. In controls, FSH declined between Days 2 and 3 and selection of DF1 occurred between Days 3 and 5. During this interval, intrafollicular estradiol concentrations increased > 5-fold in DF1, yet declined 4-fold in subordinates (p < 0.05). In DF1, total IGF-I increased 1.3-fold (p < 0.05), whereas the amounts of the 40- to 47-kDa and the 35-kDa IGFBP (ligand hybridization) decreased 2.4- and 2.5-fold, respectively (p < 0.05), compared to values in preselection follicles on Day 3; total dimeric inhibin-A decreased 1.8-fold (p < 0.05). In contrast, amounts of the 30- to 32-kDa IGFBP increased 12.4-fold (p < 0.05) in subordinates on Day 4.8 compared with preselection follicles on Day 3, while the amount of inhibins > 34 kDa decreased 4- to 9-fold (p < 0.05). In FSH-treated heifers, both selection of DF1 and atresia of subordinates were delayed by 2.2 days. Preselection follicles recovered on Day 4.9 +/- 0.1 from FSH-treated heifers were similar (p > 0.05) in almost all biochemical parameters to preselection follicles from control heifers; however, they differed markedly from both DF1 and subordinate follicles recovered from control heifers on Day 4.8 +/- 0.5. In conclusion, the decline in FSH beginning after Day 2 of the heifer estrous cycle causes differential alterations in FSH-dependent growth factors and hormones within the cohort of preselection follicles, simultaneously inducing growth and enhanced estradiol-producing capacity of the DF and atresia of subordinate follicles. PMID- 9408238 TI - A conserved Hox axis in the mouse and human female reproductive system: late establishment and persistent adult expression of the Hoxa cluster genes. AB - The mammalian female reproductive system arises from the uniform paramesonephric duct. The molecular mechanisms that establish differential development along this axis are unknown. We determined the pattern and timing of genes of the Hoxa axis in the development of the Mullerian tract. Hoxa-9, Hoxa-10, Hoxa-11, and Hoxa-13 are all expressed along the length of the paramesonephric duct in the embryonic mouse. After birth, a spatial Hox axis is established, corresponding to the postnatal differentiation of this organ system in the mouse. Hoxa-9 is expressed in the fallopian tubes, Hoxa-10 in the uterus, Hoxa-11 in the uterus and uterine cervix, and Hoxa-13 in the upper vagina. This expression pattern follows the paradigm of spatial colinearity but is a novel exception to temporal colinearity that has been considered typical of Hox genes. These genes remain expressed in the adult mouse and are expressed in the same pattern in the human. The female reproductive system undergoes dramatic structural and functional changes during the estrous cycle and in pregnancy, retaining a high degree of developmental plasticity. The late establishment of a Hox axis and persistent expression of Hox genes in the adult may play an important role in preserving this plasticity. PMID- 9408239 TI - Interaction of prostaglandin F2alpha and gonadotropin-releasing hormone on progesterone and estradiol production in human granulosa-luteal cells. AB - This study examined the effects of prostaglandin-F2alpha (PGF2alpha), GnRH, and their interactions on steroidogenesis in human granulosa-luteal cells (GLCs). Human GLCs collected from in vitro fertilization patients were cultured for one or eight days, followed by a 24-h treatment period, after which media were collected and radioimmunoassayed for progesterone and estradiol. In the first experiment, GLCs were treated with vehicle, PGF2alpha (10(-9) M), GnRH (10(-6) M), or PGF2alpha plus GnRH, with or without hCG (1 IU/ml). Neither PGF2alpha nor GnRH alone had a significant effect; however, the combination of PGF2alpha plus GnRH significantly stimulated steroidogenesis. Similarly, co-application enhanced the luteolytic effects of PGF2alpha. In a second experiment, PGF2alpha and GnRH concentration-response curves were crossed into a matrix of 49 separate treatments. Responses were plotted in three-dimensions and as two-dimensional "slices" that were analyzed statistically. In the presence of high concentrations of GnRH (10(-6) M), PGF2alpha stimulated progesterone production in a biphasic manner, as middle concentrations significantly stimulated (10(-9) M) whereas low and high concentrations did not. In the presence of middle concentrations of PGF2alpha (10(-9) M), GnRH significantly stimulated progesterone production in a linear concentration-dependent manner. Similar complexities were seen with respect to estradiol response. Thus, in the human GLC, GnRH potentiates the luteolytic effects of PGF2alpha, while it acts as a permissive factor for the luteotropic effects. Furthermore, we have revealed the complex interaction of these hormones using a three-dimensional experimental design. PMID- 9408240 TI - Small heat shock proteins and vasospasm in human umbilical artery smooth muscle. AB - Human umbilical artery smooth muscle is uniquely refractory to cyclic nucleotide dependent vasorelaxation. Small heat shock proteins (HSPs) have been implicated as contractile regulatory proteins. Thus, we hypothesized that alterations in the phosphorylation of small HSPs may contribute to human umbilical artery smooth muscle vasospasm. Physiologic contractile responses were determined in a muscle bath and compared with phosphorylation events determined with whole-cell phosphorylation and 2-dimensional gel electrophoresis. Precontraction of bovine carotid artery smooth muscle with serotonin followed by relaxation with forskolin was associated with increases in the phosphorylation of HSP27 and HSP20. Precontraction of umbilical artery with serotonin followed by forskolin treatment led to increases in the phosphorylation of HSP27. However, the umbilical artery smooth muscle did not relax, nor was there an increase in the phosphorylation of HSP20 with forskolin treatment. These data suggest that impaired cyclic nucleotide-dependent relaxation of umbilical artery smooth muscle is associated with a lack of phosphorylation of HSP20. PMID- 9408241 TI - Regulation of progesterone receptor messenger ribonucleic acid by progestin in human endometrial stromal cells. AB - Down-regulation of the progesterone receptor (PR) by its ligand has been demonstrated in breast cancer cell lines and in the rat uterus. However, in the stromal cells of endometrium, reduction of the PR level is not apparent in the luteal phase of the menstrual cycle. The purpose of this study was to determine the effect of progestin on PR and PR mRNA in isolated human endometrial stromal cells. Western blot analysis showed that progesterone or medroxyprogesterone acetate increased the two isoforms, PR-A and PR-B, in stromal cells but reduced them in glandular epithelial cells. Progestin increased the PR-A and PR-B mRNA by 2- to > 10-fold in the stromal cells of 12 specimens measured by solution hybridization-ribonuclease protection assay. A time study showed that the increase in PR mRNA required at least a 2- to 3-day incubation with progestin and that the high mRNA levels were maintained or increased slightly beyond 10 days of progestin incubation. The stimulatory effect of progestin was inhibited by RU-486 and by cycloheximide, suggesting that the up-regulation requires ligand binding to PR and de novo protein synthesis. Progestin also increased the stability of PR mRNA in endometrial stromal cells. These results demonstrated for the first time that progestin exerts an up-regulation of PR by increasing the steady-state level of PR mRNA specifically in human endometrial stromal cells. The up-regulation of PR by progestin may be mediated in part by progestin-induced endometrial stromal cell factors such as estrogen and insulin-like growth factor-I, both of which stimulated the PR-A and PR-B mRNA in stromal cells. PMID- 9408243 TI - Testis-specific expression of the mouse histone gene H1t is regulated by several promoter elements. AB - The testis-specific histone gene H1t is expressed only in mammalian testis at the stage of pachytene spermatocytes. The tissue-specific regulation of the mouse H1t gene was examined in mouse testicular primary culture cells with gene constructs consisting of H1t promoter elements fused to the chloramphenicol acetyltransferase or the firefly luciferase reporter gene. Our experiments demonstrate that expression of the mouse H1t gene is enhanced by a conserved H1 histone gene-specific TG box 452 base pairs upstream of the transcription start site. The transcription of the H1t gene appears to be reduced by sequences between -1999 and -1506. No regulatory effect could be shown for the H1 box in the expression of the mouse H1t gene. Binding of nuclear protein extracted from mouse testis to these consensus elements was shown by electrophoretic mobility shift assays with mouse testicular nuclear proteins and labeled oligonucleotides containing the upstream TG box sequences or the two testis-specific elements of the H1t gene. PMID- 9408242 TI - Role of deoxyribonucleic acid polymerase epsilon in spermatogenesis in mice. AB - Previous studies on DNA polymerase epsilon indicate that this enzyme is involved in replication of chromosomal DNA. In this study, we examined the expression of DNA polymerases alpha, delta, and epsilon during mouse testis development and germ cell differentiation. The steady-state levels of mRNAs encoding DNA polymerase epsilon and the recombination enzyme Rad51 remained constant during testis development, whereas the mRNA levels of DNA polymerases alpha and delta declined from birth until sexual maturity. Immunohistochemical staining methods, using a stage-specific model of the seminiferous epithelium, revealed dramatic differences between DNA polymerase alpha and epsilon distribution. As expected, DNA polymerase alpha and proliferating cell nuclear antigen showed relatively strong immunostaining in mitotically proliferating spermatogonia and even stronger staining in preleptotene cells undergoing meiotic DNA replication. The distribution of Rad51 was similar, but there was a dramatic peak in late pachytene cells. In contrast, DNA polymerase epsilon was detectable in mitotically proliferating spermatogonia but not in the early stages of meiotic prophase. However, DNA polymerase epsilon reappeared in late pachytene cells and remained through the two meiotic divisions, and was present in haploid spermatids up to the stage at which the flagellum starts developing. Overall, the results suggest that DNA polymerase epsilon functions in mitotic replication, in the completion of recombination in late pachytene cells, and in repair of DNA damage in round spermatids. In contrast, DNA polymerases alpha and delta appear to be involved in meiotic DNA synthesis, which occurs early in meiotic prophase, in addition to functioning in DNA replication in proliferating spermatogonia. PMID- 9408244 TI - Multiple pregnancy-associated glycoproteins are secreted by day 100 ovine placental tissue. AB - Pregnancy-associated glycoprotein (PAG)-1 (PAG1) and pregnancy-specific protein B are either identical or closely related antigens released by trophoblast binucleate cells of placentas of cattle. Sheep and other ruminants produce similar products. There is evidence, however, that these antigens, which are related structurally to the pepsinogens and other aspartic proteinases, are not single gene products but members of an extensive family. Here, the sequential use of ammonium sulfate precipitation and Sepharose blue, anion-exchange, and cation exchange chromatographies, as well as isoelectric elution from a Mono P column, has allowed several PAG1-related molecules to be purified from the medium after culture of explants from Day 100 sheep placentas. Each of these PAGs cross reacted to a varying extent with a panel of three different anti-PAG1 antisera. Four of them, all of which were major secretory products of the placenta, were subjected to amino-terminal microsequencing. Although each was related to ovine (ov) PAG1, none was identical. Reverse transcription-polymerase chain reaction was then used to amplify PAG1-related cDNA from Day 100 placental RNA. Seven novel full-length cDNA, all distinct from ovPAG1, were identified from 25 cDNA selected for sequencing. Only two of these (ovPAG3 and ovPAG7) encoded polypeptides identical in sequence at their inferred amino termini to one of the PAGs (ovPAG65) purified from explant cultures. Even so, they were only 84% identical in overall sequence. The remaining five cDNA were unique. In situ hybridization analysis revealed that expression of ovPAG3 and ovPAG7, like that of ovPAG1, is confined to trophoblast binucleate cells. The data confirm that at Day 100 of pregnancy the ovine placenta produces many different PAGs, which differ considerably in sequence and immunological cross-reactivity. PMID- 9408245 TI - Ovulation defect and its restoration by bone marrow transplantation in osteopetrotic mutant mice of Mitf(mi)/Mitf(mi) genotype. AB - Mutation within the Mitf gene causes, in microphthalmic Mitf(mi)/Mitf(mi) (mi/mi) mice, multiple defects, including white coat color and functional defects in macrophages and osteoclasts. Our previous mating experiments have demonstrated that the mi mutation reduces the numbers of newborns and induces uterine inversion at delivery. The present study was designed to determine the causes of these pregnancy defects. The histology and number of F4/80-positive macrophages were not different between the ovaries of 23-day-old mi/mi and +/+ mice given eCG 48 h earlier. When ovulation was induced in these mice by hCG, the number of ovulated ova was significantly smaller in mi/mi mice than in wild-type (+/+) mice (p < 0.05). When bone marrow cells from +/+ mice were transplanted i.p. into 42 mi/mi female newborns, successful transplantation was observed in 16 of them at 20 days after birth. In one of these, the upper incisors had erupted. The mean number of tubal ova in mi/mi mice significantly increased after transplantation (p < 0.05) and was almost equal to that of +/+ mice. No uterine inversion occurred at 6 deliveries in 5 mi/mi females after bone marrow transplantation, while it occurred at 4 of 5 deliveries in mi/mi females during the same observation period (p < 0.05). These results indicate that bone marrow-derived cells, defective in mi/mi mice, are necessary for normal ovulation and delivery; the findings are consistent with the notion that macrophages play major roles in ovulation. PMID- 9408246 TI - Fluorometric assessments of mitochondrial function and viability in cryopreserved bovine spermatozoa. AB - Mitochondrial function and sperm viability were quantified in samples of cryopreserved bovine spermatozoa from 12 bulls using fluorometric techniques. The active mitochondria of the spermatozoa were fluorescently stained using three different fluorophores: rhodamine 123 (R123), 5,5',6,6'-tetrachloro-1,1',3,3' tetraethylbenzimidazolyl-carbocyan ine iodide (JC-1) or MitoTracker Green FM (MITO). The stained spermatozoa, and companion aliquots that had been stained with SYBR-14 (a living-cell nucleic acid stain) and propidium iodide to assess viability, were quantified using flow cytometry. The resulting fluorescent measurements of mitochondrial function were compared with microscopic assessments of progressive sperm motility immediately after thawing, with motility after 3-h incubation at 37 degrees C, and with the fluorescent assessment of sperm viability. Staining with either R123 or MITO resulted in a single green population. In contrast, the JC-1 staining of mitochondria produced both green and red-orange populations of spermatozoa and sometimes a progressive gradient between the two populations. The ability of JC-1 to discriminate between mitochondria exhibiting high membrane potential from those having low to medium membrane potential provided a more rigorous estimate of metabolic function than the other two fluorescent stains. Overall, the three fluorometric measurements of mitochondrial function were highly correlated with each other, with the SYBR-14 assessment of viability, and with the microscopic estimates of motility. PMID- 9408247 TI - Temperature-dependent hyperactivated movement of hamster spermatozoa. AB - Immotile hamster spermatozoa from the caudal epididymis were activated in physiological medium at a temperature range between 22 and 40 degrees C. With rising temperature, the sperm swimming velocity and flagellar beat frequency increased, but the flagellar bend angles and waveforms did not change dramatically. All spermatozoa exhibited a symmetric and progressive movement at this temperature range. Hyperactivation was induced by incubating activated spermatozoa in vitro at 37 degrees C for 3.5 h under 5% CO2 in air. When examined at 37 degrees C, spermatozoa expressed a hyperactivated motility characterized by asymmetric and circular flagellar beating with large principal and reverse bends. When the temperature was lowered to 33 degrees C, the sperm motility pattern characteristic of hyperactivation became less prominent. In particular, the reverse bend originating in the flagellar middle piece did not propagate to the principal piece. However, normal hyperactivated movement was restored as the temperature was raised to 37 degrees C, indicating that in vitro expression of hyperactivated motility of hamster spermatozoa is a temperature-dependent phenomenon. To further investigate temperature effects on the flagellar motor apparatus, spermatozoa were demembranated and then reactivated with ATP. In contrast to membrane-intact hyperactivated spermatozoa, demembranated spermatozoa exhibited a hyperactivation-like motility independent of the temperature range tested (22-40 degrees C). This strongly suggests that temperature regulates initiation and maintenance of hyperactivated motility and that its effect is mediated by the plasma membrane. PMID- 9408248 TI - Glutathione oxidation is associated with altered microtubule function and disrupted fertilization in mature hamster oocytes. AB - We hypothesized that depletion of glutathione (GSH) with diamide, a relatively specific GSH oxidant, may alter the meiotic spindle apparatus in mature hamster oocytes. Immunofluorescent analysis of oocytes exposed to diamide for 1.5 or 3 h revealed time- and concentration-dependent disruption of spindle morphology accompanied by chromosome clumping. In oocytes first cultured in diamide for 1.5 h and then in diamide-free medium for 1.5 or 3 h, microtubules appeared to repolymerize, but normal spindle structure was not regained. HPLC confirmed that diamide oxidized oocyte GSH under conditions identical to those associated with spindle-related abnormalities. Exposure of oocytes to 25 or 50 microM diamide before in vitro fertilization did not affect their ability to undergo fertilization. A significant proportion of the fertilized oocytes that had been exposed to 50 microM diamide before insemination exhibited abnormal multiple female pronuclei with an apparently normal male pronucleus. These observations indicate that mature hamster oocytes are susceptible to oxidative stress during the critical period that precedes fertilization and provide further evidence that GSH plays important roles in oocyte spindle function and pronucleus development. PMID- 9408249 TI - Glutathione synthesis during in vitro maturation of bovine oocytes: role of cumulus cells. AB - Glutathione (GSH) synthesis during in vitro maturation (IVM) has been shown to play an important role in embryo development. The present study was carried out to evaluate the role of cumulus cells in GSH synthesis during IVM of bovine oocytes in the presence of cystine, cysteine, the cysteine analogue N acetylcysteine, and cysteamine. For this purpose, cumulus-oocyte complexes (COCs), denuded oocytes (DOs), and DOs in coculture with a cumulus cell monolayer were used. An increase in GSH level stimulated by cystine was observed only in the presence of cumulus cells, either with COCs or in DOs matured on a coculture monolayer. Addition of cysteine and cysteamine to IVM medium increased GSH levels in COCs and DOs. N-Acetylcysteine increased GSH levels only in DOs. Moreover, cumulus cells contributed to the stimulatory effect exerted by cysteine and cysteamine on GSH synthesis in COCs. These results indicate that cumulus cells during IVM play an important role in oocyte GSH synthesis, allowing the oocytes to use cystine and contributing to the stimulatory effect exerted by cysteine and cysteamine. In addition, these results demonstrate that IVM medium supplemented with cysteine or cysteamine increased GSH content in oocytes without cumulus mass (DO) and in the absence of a cumulus cell monolayer. This may be useful to increase the efficacy of IVM of those oocytes having few cumulus cell layers, in a system without coculture. PMID- 9408250 TI - Analysis of the protein composition of the mouse sperm perinuclear theca and characterization of its major protein constituent. AB - The perinuclear theca (PT) is a cytoskeletal structure that covers the nucleus of mammalian spermatozoa and is believed to have a membrane-binding role. The objectives of this study were to analyze the protein composition of the mouse PT, to identify and sequence its major protein component, and to characterize this protein's transcriptional and translational origins during spermatogenesis. The PT was extracted from demembranated and acrosome-depleted mouse sperm heads by alkaline treatment. The protein profile of the PT extract was composed of several polypeptides, of which a 15-kDa subacrosomal protein predominated and was found to be immunocross-reactive with a previously cloned 15-kDa PT protein of the rat (PERF 15) that belongs to a family of lipid-binding proteins. A primer pair designed from rat PERF 15 cDNA was then used to screen a mouse testicular cDNA library by polymerase chain reaction (PCR). The deduced amino acid sequence obtained from the PCR product was almost identical to the testicular-specific rat PERF 15. Developmental Northern blots and in situ hybridization studies performed with riboprobes encoding the mouse PERF 15 cDNA revealed that mRNA levels were highest in round and early elongating mouse spermatids. Immunohistochemistry indicated that PERF 15 began to be expressed in the cytoplasm of mid-pachytene spermatocytes and appeared to reach maximum expression in the distal cytoplasm of late elongating mouse spermatids, long after transcriptional arrest. During the development of round and early elongated spermatids, the immunolabel became progressively concentrated over the anterior half of the spermatid nucleus, suggesting a subacrosomal deposition of PERF 15 during this phase of mouse spermiogenesis. PMID- 9408251 TI - A novel steroid inhibitor for a rat 20alpha-hydroxysteroid dehydrogenase isozyme. AB - 20alpha-Hydroxysteroid dehydrogenase (20alpha-HSD, E.C.1.1.1.149) in rat luteal tissue, which catalyzes conversion of progesterone to a biologically inactive steroid, 20alpha-hydroxypregn-4-ene-3-one (20alpha-OHP), suppresses progesterone secretion into the circulation. An increase in 20alpha-HSD activity in luteal tissue in rats is a prerequisite for functional corpus luteum regression. This study was undertaken to find a steroid inhibitor for ovarian cytosolic 20alpha HSD activity among derivatives based on progesterone structure. A derivative designated as STZ26 (D-homo-16-oxa-4-androstene-3,16alpha-dione) was found to inhibit potently 20alpha-HSD activity in cultured luteal cells. Ovarian 20alpha HSD activity consists of two isoforms (HSD1 and HSD2). Kinetic analyses of STZ26 for HSD1 and HSD2 showed that the compound suppressed only HSD1 activity by competitive inhibition. Pseudopregnant rats were treated with STZ26 from 13 to 19 days after cervical stimulation. Either an elevation of peripheral 20alpha-OHP levels or a concomitant depletion of peripheral progesterone levels at the end of pseudopregnancy was considerably inhibited in treated animals, although not completely. The results showed that STZ26 is a biologically active inhibitor for HSD1 activity in the luteal tissue and suggested that the depletion of progesterone levels toward the end of pseudopregnancy is not solely due to the elevation of HSD1 activity. PMID- 9408252 TI - Interleukin (IL)-6 and IL-8 production by human amnion: regulation by cytokines, growth factors, glucocorticoids, phorbol esters, and bacterial lipopolysaccharide. AB - Amniotic fluid at term contains high concentrations of interleukin (IL)-6 and IL 8. The source of these cytokines has not been identified, although the fetal membranes (amnion and chorion) are likely contributors. Amnion cytokine production was investigated by using amnion cells isolated by enzymatic digestion (from placentas delivered at term before labor) and cultured in vitro. IL-6 and IL-8 were measured in conditioned media by ELISA. Amnion cells produced detectable amounts of both IL-6 and IL-8 throughout the 7-day culture period. The ratio of IL-8 to IL-6 was approximately 5:1, similar to the ratio found in amniotic fluid. Production of both IL-6 and IL-8 was stimulated in a concentration-dependent fashion by interleukin-1beta (0.1-10 ng/ml), tumor necrosis factor-alpha (1-100 ng/ml), and bacterial lipopolysaccharide (0.1-10 microg/ml), and also by 100 nM phorbol 12-myristate 13-acetate. Epidermal growth factor (1-25 ng/ml) had only minimal effects on amnion cytokine production. Dexamethasone (10 nM) inhibited IL-6/-8 production by approximately 50% throughout the culture period. Production of IL-6/-8 by cultured amniotic fibroblasts, which under basal conditions was much lower than that by epithelial cells, was regulated by all the agents tested in a fashion similar to that of the epithelial cells. These findings suggest that the amnion contributes to the pool of IL-6 and IL-8 in amniotic fluid. We speculate that amnion-derived cytokines might have functions during normal human parturition that are distinct from their conventional roles as inflammatory mediators. PMID- 9408253 TI - Bovine endometrial retinol-binding protein secretion, messenger ribonucleic acid expression, and cellular localization during the estrous cycle and early pregnancy. AB - Retinol-binding protein (RBP) mRNA was localized to luminal and glandular epithelial cells of bovine endometrium by in situ hybridization. Relative levels of endometrial RBP mRNA expression during the estrous cycle and early pregnancy were determined by quantitative slot blot analysis and contrasted with uterine luminal concentrations of RBP. Expression of mRNA was moderate at Day 1 after estrus, declined at Day 5, reached lowest levels by Day 10, rose significantly through Days 15-17, and peaked at Day 20. RBP mRNA expression in pregnant animals was similar to that in cyclic animals on Day 15, doubled between Days 17 and 20, remained constant through Day 22, and rose slightly thereafter. Luminal RBP concentrations of cyclic cows, as determined by ELISA, decreased from Day 1 through Day 10, rose dramatically on Day 15, then declined through Day 20. Concentrations of RBP in uterine flushes from pregnant animals were similar to those of cyclic cows on Day 15 but remained relatively constant through Day 17. It is concluded that 1) RBP synthesis occurs in the luminal and glandular epithelial cells, 2) RBP transcription and secretion are correlated with each other, and 3) ovarian steroids, possibly in conjunction with uterine concentrations of their receptors, modulate uterine RBP expression. PMID- 9408254 TI - Gonadotropin versus steroid regulation of the corpus luteum of the rhesus monkey during simulated early pregnancy. AB - During the nonconceptive cycle in primates, progesterone is a likely intermediary for several LH-dependent events in the ovary including ovulation, luteinization of the follicle wall, and maintenance of the developed corpus luteum. To determine whether progesterone is an important local factor in the ability of chorionic gonadotropin (CG) to enhance luteal structure and function in early pregnancy, rhesus monkeys received hCG in a dose-escalating regimen (15-2880 IU twice daily) beginning on Day 9 of the luteal phase of the natural menstrual cycle to simulate the rapid rise in serum CG levels associated with early pregnancy. Some animals were concomitantly treated with the 3beta-hydroxysteroid dehydrogenase (3beta-HSD) inhibitor trilostane (500 mg twice daily) to suppress progesterone production during gonadotropin stimulation. Corpora lutea were removed after 1, 3, 6, and 9 days of treatment (n = 3-4 per group); time-matched control tissues were obtained from untreated animals (n = 3 per group). Treatment with hCG prevented both the decrease in luteal wet weight (p < 0.05) and the histologic indices of luteal regression seen in controls during the menstrual cycle. However, coadministration of the progesterone synthesis inhibitor led to early declines in luteal wet weight (p < 0.05) and luteal cell size compared to treatment with hCG alone. Luteal progesterone receptor (PR) mRNA content increased (p < 0.05), but the percentage of cells staining positive for immunoreactive PR declined (p < 0.05) over the treatment interval in all groups. CG administration alone and in combination with trilostane increased PR staining intensity in some luteal cells within 1 day of treatment; intensely staining cells persisted around vascular elements after 9 days of treatment with hCG+trilostane but not with hCG alone. These data suggest that some, but not all, actions of CG to maintain the primate corpus luteum in early pregnancy are mediated by progesterone via a receptor-mediated pathway. PMID- 9408255 TI - Localization of lipid peroxidation-derived protein epitopes in the porcine corpus luteum. AB - Reactive oxygen species (ROS) generated within the corpus luteum (CL) are believed to play an integral role in luteolysis. Unsaturated lipids are susceptible to peroxidation by ROS, leading to the formation of toxic aldehydes such as malondialdehyde (MDA) and 4-hydroxy-nonenal (4-HNE) that can attack cellular and extracellular proteins. Aldehyde conjugation of proteins can have diverse effects on cell function, including cross-linking of cell surface receptors, alteration of enzyme activities, and modification of lipoproteins. The objectives of the present study were to 1) determine by immunological techniques whether lipid peroxidation-derived protein epitopes (i.e., MDA- and 4-HNE-lysine) could be detected in situ during the natural life span of the CL, and 2) determine the temporal and spatial localization of these epitopes. Fresh luteal tissue was collected from young gilts at distinct phases of the estrous cycle: early (Days 4-7), mid (Days 8-12), and late (Days 14-18). Immunocytochemistry was performed on tissue sections using monoclonal antibodies against MDA-lysine and 4 HNE-lysine. Positive immunostaining was evident in porcine CL at all stages of the estrous cycle. Immunoactivity was associated primarily with the steroidogenic cells, most consistently with the large luteal cells, and was most intense in regressing tissue (i.e., late). No staining was observed in sections incubated with nonimmune mouse serum or when anti-4-HNE-lysine was preadsorbed with 4-HNE protein preparations. Immunoblotting of protein extracts from luteal tissue revealed three major reactive bands. At least two of these bands were closely associated with extracted oxidized human low-density lipoprotein (LDL) and with 4 HNE-conjugated human apoprotein B. These results demonstrate the presence of lipid peroxidation-derived protein epitopes within the porcine CL and show that one or more of these may be an oxidized-LDL moiety. This represents the first direct in situ evidence that protein modification by lipid peroxidation products occurs during the natural luteal life span. The physiological consequences remain to be determined. PMID- 9408256 TI - A comparison of the effects of testicular maturation and aging on the stage specific expression of CP-2/cathepsin L messenger ribonucleic acid by Sertoli cells of the Brown Norway rat. AB - We have examined the effects of maturation and aging on expression of cyclic protein-2 (CP-2)/cathepsin L mRNA by rat Sertoli cells. During maturation (25-68 days of age) there was a significant, 7.6-fold increase in CP-2/cathepsin L mRNA. There was a significant, 2.9-fold increase in SGP-2 mRNA during the same period. In situ hybridization analysis demonstrated that stage-specific expression of CP 2/cathepsin L mRNA developed between 30 and 35 days. By 64 days of age, this transcript was only detectable in Sertoli cells within stage VI-VII tubules. Northern blot analysis of 23-mo-old testes revealed statistically significant decreases in total testis contents of CP-2/cathepsin L and SGP-2 mRNAs (4- and 3 fold, respectively) in testes that lacked germ cells but not in aged testes with a full complement of germ cells. In situ hybridization analysis revealed high amounts of CP-2/cathepsin L mRNA in all aged tubules that were losing germ cells via apoptosis, regardless of the stage of the cycle. Immunocytochemical analysis demonstrated that in these aged, regressing tubules, CP-2/cathepsin L protein became concentrated in dying or dead germ cells. PMID- 9408257 TI - Amino acids in maturation medium and presence of cumulus cells at fertilization promote male pronuclear formation in porcine oocytes matured and penetrated in vitro. AB - The present study was conducted to examine the ability of porcine oocytes to achieve male pronuclear (MPN) formation when they are matured and penetrated in vitro under various culture conditions. When cumulus-enclosed oocytes were cultured for 24-48 h in modified Whitten's medium (pH 7.4) supplemented with 10% porcine follicular fluid, 10 IU eCG/ml, and 10 IU hCG/ml (designated mWM-FG), nuclear maturation of oocytes reaching metaphase II was completed by 36 h after the start of culture. However, there were no differences in the proportions (94 95%) of oocytes penetrated in vitro by cryopreserved ejaculated spermatozoa or in the rates (35-45%) of MPN formation between oocytes cultured for 36 and 48 h. When cumulus-enclosed oocytes were cultured for 36 h in mWM-FG supplemented with 2% (v:v) minimal essential medium (MEM) essential amino acids (EAA) with the addition of 0.1 mM glutamine and/or 1% (v:v) MEM nonessential amino acids (NEAA) and inseminated in vitro, 93-97% of oocytes were penetrated regardless of the presence of amino acids during maturation, but the rates of MPN formation were higher in the presence (79-84%) than in the absence (51%) of any amino acids. The addition of EAA+NEAA and/or 0.57 mM cysteine to mWM-FG also did not affect sperm penetration in vitro, while it promoted MPN formation (76-83%) in penetrated oocytes as compared with those matured in the absence of amino acids and cysteine (53%). When oocytes were freed from cumulus cells after culture in mWM-FG, sperm penetration rates were not different between cumulus-enclosed (100%) and cumulus free (92%) oocytes, but the rate of MPN formation was higher in cumulus-enclosed (53%) than in cumulus-free (28%) oocytes. When EAA+NEAA+cysteine was added to mWM FG, MPN formation was not improved in cumulus-free oocytes but was much improved (78%) in cumulus-enclosed oocytes. These results indicate that MPN formation in porcine oocytes is promoted by the addition of amino acids and/or cysteine in simple maturation medium and by the presence of cumulus cells at fertilization in vitro. PMID- 9408259 TI - Measurement of inhibin and activin in early human pregnancy: demonstration of fetoplacental origin and role in prediction of early-pregnancy outcome. AB - To determine the source of the dimeric glycoproteins inhibin A (alpha-betaA) and activin A (betaA-betaA) in early pregnancy, we analyzed serial blood samples from women who became pregnant following in vitro fertilization (IVF) with fresh embryo transfer (ET; n = 52) and from women who achieved pregnancy with frozen thawed embryos (n = 8). Elevated serum levels of inhibin A were detected in ongoing pregnancies from 4 wk gestation (13 days following ET) and increased to an initial peak at 9-10 wk gestation. Significantly higher levels (p < 0.05) were found in the multiple pregnancies, and nonviable clinical pregnancies had very low levels of inhibin A. Total activin A was detectable 14 days after ET (positive pregnancy test), and higher levels were associated with multiple gestations while rapidly falling levels heralded embryonic demise. The fetoplacental unit is thus confirmed as the major source of these glycoproteins. Inhibin pro-alphaC, which circulates in great excess as a functionally inactive monomer and as part of high molecular weight functional dimers, was detectable at levels above normal late-luteal values in singleton and multiple IVF arising from fresh ETs. With frozen-thawed embryo pregnancies, the levels of pro-alphaC containing inhibins were extremely low, confirming that the corpus luteum of pregnancy is the major source of the alpha monomer. The initially low levels and very rapid decline in inhibin A in pregnancies with embryonic failure suggest a role for this glycoprotein as a monitor of early-pregnancy viability. PMID- 9408258 TI - Stimulatory effect of growth hormone on in vitro maturation of bovine oocytes is exerted through the cyclic adenosine 3',5'-monophosphate signaling pathway. AB - The aim of this study was to investigate whether the stimulatory effect of growth hormone (GH) on the in vitro maturation and cumulus expansion of bovine oocytes is exerted through the cAMP or the tyrosine kinase pathway. Therefore bovine cumulus-oocyte complexes (COCs) were cultured in Medium 199 without fetal calf serum and gonadotropins, but supplemented with 100 ng/ml bovine GH (bGH; NIH-GH B18) with or without 10 microM methyl 2,5-dihydroxycinnamate (erbstatin analogue), a specific tyrosine kinase inhibitor; 100 microM 2',3' dideoxyadenosine (DDA), a specific adenylate cyclase inhibitor; or 10 microM H 89, a specific inhibitor of cAMP-dependent protein kinase A. Epidermal growth factor (EGF; 20 ng/ml) was added as a positive control for tyrosine kinase activation, and FSH (0.05 IU/ml) was added as a positive control for cAMP mediation during in vitro maturation in the absence or presence of the inhibitors. Culture was performed at 39 degrees C in a humidified atmosphere with 5% CO2 in air. To assess the effect on nuclear maturation, the proportion of oocytes in metaphase II stage after 16 h of culture was determined using 4,6 diamino-2-phenylindole staining. To determine the effect on cumulus expansion, the diameter of COCs at the onset and after 24 h of culture was measured. The stimulatory effects of GH on oocyte maturation and cumulus expansion were blocked by DDA and H-89 (p < 0.01). Similarly, FSH-induced cumulus expansion was abolished by DDA and H-89 (p < 0.05), while DDA did not block either EGF-induced oocyte maturation or cumulus expansion. Erbstatin analogue significantly blocked the stimulation of oocyte maturation and cumulus expansion by EGF (p < 0.02) but did not inhibit GH action on the COCs. It is concluded that the stimulatory effect of GH on oocyte maturation and cumulus expansion is mediated by the cAMP signal transduction pathway and not by JAK2 phosphorylation. PMID- 9408260 TI - Intracytoplasmic sperm injection of in vitro-matured equine oocytes. AB - Intracytoplasmic sperm injection (ICSI) was performed on equine oocytes matured in vitro. The oocytes were aspirated from abattoir ovaries and matured in vitro for 36 h at 38 degrees C. ICSI was performed using frozen/thawed stallion semen after swimup in medium containing human serum albumin. Sperm-injected oocytes were either 1) cultured in vitro for 10, 20, or 72 h; 2) transferred to oviducts of pseudopregnant mice; or 3) transferred to a synchronized mare after initial in vitro culture. The transferred ova were recovered after 72 h, and all ova were subsequently fixed, stained, and processed for light and transmission electron microscopy. Single pronucleus formation was observed in 2 out of 12 presumptive zygotes 10 h postinjection, at which time abundant cortical granules were observed in the subplasmalemmal region. Twenty hours postinjection, however, 2 pronuclei were observed in 6 of 12 injected oocytes (fertilization rate 50%), and almost all cortical granules were released. The cleavage rate in vitro was 16% after 72 h in culture, and the most advanced embryo stages obtained were 6- to 8 cell embryos. The cleavage rate in vivo was very low since only 1 of 10 recovered had cleaved to the 2-cell stage. Thus, in conclusion, ICSI fertilization of equine oocytes did result in fertilization, pronucleus formation, and cortical granule release. However, the observed fertilization rate and oocyte activation was not paralleled by substantial cleavage of the zygotes. PMID- 9408261 TI - Distribution and possible novel role of phospholipid hydroperoxide glutathione peroxidase in rat epididymal spermatozoa. AB - The selenoenzyme phospholipid hydroperoxide glutathione peroxidase (PHGPx, EC 1.11.1.12) is present, in both free and membrane-bound form, in several mammalian tissues. It utilizes thiols such as glutathione to specifically scavenge phospholipid hydroperoxides. The testis exhibits the highest PHGPx-specific activity so far measured, and interest in the presence and function of the enzyme in this tissue has recently grown. Here we report the localization of PHGPx in rat epididymal spermatozoa and its distribution in subfractions obtained by sucrose density gradient centrifugation. Immunochemical evidence and enzymatic activity revealed for the first time that PHGPx is present in sperm heads and tail midpiece mitochondria. The binding of the enzyme to spermatozoa, head, and mitochondria was barely affected by ionic strength or thiols or detergents, as compared to the detachment of PHGPx obtained from testis nuclei. Moreover, we demonstrated that pure PHGPx exhibits a higher thiol-oxidase activity toward isolated epididymal caput protamines than toward protamines from epididymal cauda. These results suggest a role for the enzyme in the maturation of spermatozoa through the metabolism of hydroperoxides and sperm thiol oxidation, in addition to its serving as an antioxidant protector. PMID- 9408262 TI - Influence of dehydroepiandrosterone on the expression of insulin-like growth factor-1 during cystogenesis in polycystic rat ovaries and in cultured rat granulosa cells. AB - This study was designed to investigate the expression of insulin-like growth factor-1 (IGF-1) during cystogenesis in the dehydroepiandrosterone (DHEA)-induced rat polycystic ovarian syndrome (PCO) model. IGF-1 expression patterns in DHEA treated rat ovaries were compared with those in control ovaries. In situ hybridization revealed a similar distribution of IGF-1 mRNA in DHEA-treated and control ovaries: in both, IGF-1 mRNA expression was confined to the granulosa cells of preantral and small antral follicles. Some hybridization signals for IGF 1 mRNA were also found in theca and infrequently in the interstitial cells. No signal was observed in larger antral follicles, atretic follicles, or cysts. This similarity indicates that there might be a shared mechanism in the early follicular development of normal folliculogenesis and DHEA-induced cystogenesis. The effects of DHEA on granulosa cells were analyzed in vitro in their quiescent, proliferative, differentiative, and preovulatory stages. Northern analysis revealed three transcripts for IGF-1 (7.5 kilobases [kb], 1.6 kb, and a group of signals between 0.4 and 0.9 kb) in cells at all stages except the preovulatory. The strongest signal was observed in cells of the proliferative stage of control cultures, while expression of IGF-1 increased only in the DHEA-treated cells cultured in the differentiative stage (when they secrete estrogen). Increase in IGF-1 expression may contribute to the hypersteroidogenism observed in the DHEA treated rat PCO model. PMID- 9408263 TI - Subcellular localization of the regulatory subunits of cyclic adenosine 3',5' monophosphate-dependent protein kinase in bovine spermatozoa. AB - Cyclic AMP (cAMP) is a regulator of sperm flagellar activity. The action of this cyclic nucleotide is presumably mediated by cAMP-dependent protein kinase (PKA). PKA is localized or targeted to specific subcellular sites through the interaction of PKA regulatory subunits with A-kinase anchoring proteins (AKAPs). We have recently shown that the addition of PKA anchoring inhibitor peptides to spermatozoa leads to the complete arrest of motility. A knowledge of the subcellular localization of PKA and AKAPs is essential for an understanding of how cAMP acts in spermatozoa. In this report, monospecific, affinity-purified, antipeptide antibodies were used to determine the distribution of the regulatory (R) subunit isoforms. Immunocytochemistry staining revealed that RIalpha and RIbeta subunits are both localized predominantly in the acrosomal segment of the head, although they have distinct staining patterns within this region. In addition to the head, RIbeta was observed in the midpiece of the tail while RIalpha was detected in the connecting piece. RIIalpha is prominent in the axonemal region of the flagellum but was not observed in the head region. These data suggest distinct roles for each of these isoforms in sperm functions such as motility and the acrosome reaction. PMID- 9408264 TI - Prostaglandin G/H synthase-2 is expressed in bovine preovulatory follicles after the endogenous surge of luteinizing hormone. AB - To determine whether prostaglandin G/H synthase-2 (PGHS-2) induction occurs under physiological conditions, heifers were ovariectomized and preovulatory follicles were isolated 0 (n = 4), 18 (n = 4), and 24 h (n = 4) after the onset of estrus. The time of the LH surge was determined in blood samples collected every 4 h pre- and postestrus. Preparations of follicle wall and isolated theca interna and granulosa cells were analyzed for PGHS-2 protein and mRNA by immunoblot and Northern blot, respectively. Immunohistochemistry was used to document the in situ localization of PGHS-2 protein, and follicular fluid concentrations of prostaglandin (PG) E2 and PGF2alpha were determined to monitor changes in PG synthetic activities. Results showed that PGHS-2 mRNA (4.0 kilobases) and protein (74000 Mr) were absent in preovulatory follicles isolated at the onset of estrus, low in follicles obtained 18 h after estrus (16.0 +/- 1.2 h post-LH surge), and markedly induced 24 h postestrus (20 +/- 0 h post-LH surge). Immunoblot and immunohistochemical analyses revealed that PGHS-2 protein was selectively induced in granulosa cells. Follicular fluid concentrations of PGE2 and PGF2alpha increased significantly (p < 0.01) between 0 and 24 h after estrus (from 2.8 +/- 0.2 to 87.9 +/- 30.9 ng/ml for PGE2; from 0.05 +/- 0.02 to 68.9 +/- 23.6 ng/ml for PGF2alpha). Collectively, these results clearly demonstrate that the induction of PGHS-2 in bovine preovulatory follicles is a physiological event that occurs after the endogenous LH surge. PMID- 9408265 TI - Investigation of the thoracic electrical impedance during endotoxemia in dogs. AB - The purpose of our study was to examine the effect of sustained low-dose endotoxin infusion on changes in thoracic electrical impedances (Z0 at end expiratory apnoea, Zmax at end-inspiration), Zmax-Z0, hematocrit (H) values, and extravascular lung water (EVLW) estimated with gravimetrical analysis (WW/DW) and the impedance method (dEVLWimp) in anesthetized dogs. To define the role of H in changes in the Z0 and Zmax, we applied splenectomy. To determine whether changes in respiration might be involved in changes in Z0 and Zmax, mechanically ventilated dogs were examined. During the infusion of endotoxin there was an increase in Z0, Zmax, H and the respiratory rate in the spontaneously breathing dogs. In splenectomized dogs both impedances slightly increased without any significant change in H. In the ventilated dogs Z0 and Zmax increased similarly, and H increased to a slightly lower level than in the spontaneously breathing dogs. In the ventilated-splenectomized dogs no changes were found in the impedances and in H. The EVLW values showed no serious edema in the endotoxemic groups. The results suggest that Z0 increased mainly in association with the increase in H. The Zmax-Z0 parameter proved to be a suitable parameter for demonstrating the changes in respiration. PMID- 9408266 TI - Technique and results of duodenum-preserving resection of the head of the pancreas in the treatment of chronic pancreatitis. AB - Surgical treatment of chronic pancreatitis is either by ductal decompression or resection of the pancreas. Among various resection operations the duodenum preserving resection of the head of the pancreas is the newest surgical technique. At the 2nd Dept. of Surgery of Debrecen Medical University duodenum preserving resection of the head of the pancreas with simple modified reconstruction was performed in 22 patients with chronic pancreatitis. The authors evaluate the early and late results: there was no mortality. The rate of complication was 27.2%. Considering late results complete relief of pain was found in about 80% of patients. The authors suggest that this type of resection can be well applied in the treatment of chronic pancreatitis with low risk and relatively good late results. PMID- 9408268 TI - Frequency of locoregional and axillary recurrences among patients following breast cancer operations. AB - Our department performed 554 operations for breast cancer between 01. 01. 1986 and 31. 12. 1995. When following up these 554 patients, we recognized local recurrences, axillary and neck-region lymph node recurrences in 32 cases (5.8%). Breast conserving procedure was introduced at our department within the period above. What our experience proves is, that the raise in the frequency of breast conserving procedures did not make the rate of locoregional recurrences grow. Together with applying breast conserving procedures we extended the application of axillary block dissections, in terms of indications, and also of numbers of lymph nodes removed. The past five years axillary recurrences did not appear among our patients. PMID- 9408267 TI - Operative hysteroscopy: minimally invasive surgery to control of menorrhagia. AB - Diagnostic hysteroscopy is a valuable method for evaluation of intrauterine disorders. After diagnosing, the endoscopic treatment of these pathologies is the major question of past decade. Possibility of solving cause of infertility or abnormal uterine bleeding without laparotomy or hysterotomy/hysterectomy is the great advantage of operative hysteroscopic methods. In Department of Obstetrics and Gynaecology of University Medical School of Debrecen more than 1400 hysteroscopic interventions were performed from 1 September 1989 to 31 December 1996. In treatment of intractable uterine bleeding 347 operative hysteroscopy (targeted biopsy, polypectomy, transcervical endometrial ablation, fibroid resection etc.) were performed. The rate of complications was low, only 2% (4 perforations and 2 bleedings). The high success rate and low rate of complications offers a modern, safe, minimally invasive method for treatment of menorrhagia. PMID- 9408269 TI - Penetrating keratoplasty for pseudophakic bullous keratopathy. AB - Corneal decompensation after cataract surgery and intraocular lens (IOL) implantation has become the leading indication for penetrating keratoplasty during the past decade. We reviewed the clinical course and the surgical management of 212 patients with penetrating keratoplasty for pseudophakic bullous keratopathy treated at our Department during the last 15 years. Corneal transplantation for pseudophakic bullous keratopathy was successful in 76%. One third of the patients achieved a long-term visual acuity of 0.5 and more. At keratoplasty the original IOL was left in place in 129 eyes. We removed from and not replaced the IOL in 37 eyes and we performed IOL exchange in 46 eyes. The secondary IOL was an angle-supported flexible anterior chamber lens in 40 cases, and a suture fixated posterior chamber lens in 8 cases. During the penetrating keratoplasty for pseudophakic bullous keratopathy the most important question is how to manage the previously implanted intraocular lens. We have to decide whether the intraocular lens should be removed or replaced at the time of surgery. The choice of removing, retaining or replacing the intraocular lens at keratoplasty depends on the variable intraocular pathological conditions and each case requires individual evaluation. PMID- 9408270 TI - Scleral reinforcement in progressive myopia and intraoperative ultrasound control of the cadaver fascia lata strip. AB - Scleral reinforcement (sustentaculum sclerae) is one of the operations against myopia. During the progression of myopia the eye grows. The chorioretinal layer can only moderately follow the expansion of the sclera, and mostly this is the cause of the complications that can cause permanent visual acuity decrease. The aim of the operation is to strengthen the posterior part of the sclera by implantation of a cadaver fascia lata strip, that slows down or stops the expansion of the eyeball, and this way prevents the development of complications. At our Department we perform the Snyder-Thompson technique. Between 1984 and 1994 343 operations were performed at our Department. We followed up the changes of the axial lengths by ultrasound (A-mode) examinations. The axial length decreased in 43.7% remained unchanged in 22.2% and increased in 34.1% of the cases. We controlled the location of the strip intra- and postoperatively with B-mode ultrasound. Nowadays scleral reinforcement is still the only possibility to stop or slow down the expansion of the eyeball in cases of progressive high myopia. PMID- 9408271 TI - Pancreatic pseudocyst propagating into retroperitoneum and mediastinum. AB - The extrahepatic pseudocysts of pancreatic origin sometimes propagate into mediastinum and retroperitoneum. A large pseudocyst of pancreatic origin propagating into the mediastinum up to the aortic arch is published. The surgical intervention has been urged by the dislocation of the heart, by the danger of autodigestion of the mediastinal aorta and by the danger of rupture. Attention is directed to the external drainage operation which is suitable for emptying of the pseudocyst not independently from the ripeness of pseudocyst wall and the characteristics of the pseudocyst content. Should the time short after the exacerbation and should the amylase content high in the cyst, the immediate result of external drainage better and reliable safe. PMID- 9408272 TI - Transabdominal preperitoneal herniorraphy: technique and results. AB - As enthusiasm for laparoscopic surgery has grown, laparoscopic approaches to the groin hernia have evolved. The most widely accepted laparoscopic repair employs the placement of a large sheet of mesh in a preperitoneal position to cover potential hernia spaces. Between March 1994 and February 1997 160 inguinal and 3 femoral hernia were operated of an transabdominal preperitoneal (TAPP) polipropylen mesh. 131 patients were operated (128 males and 3 females, ranging in age from 19 to 82 years), 31 (23%) of them had bilateral hernias. Recurrent hernia was the indication in 52 (32%) cases. Average operating time for unilateral repair was 80 minutes and for bilateral repairs was 108 minutes. Postoperative complications included 7 (4.3%) cases of transient neuralgias, 20 (12%) cord/scrotal transient seromas-hematomas and 2 (1.2%) hydrocele. The 5 (3.1%) early recurrences were considered to be caused by technical inexperience and/or too small prosthetic patch. The laparoscopic hernioplasty has definitive advantage: minimal postoperative pain, short hospital stay (average postoperative time of hospitalization 3.1 days) and early restoration of full physical activity (in 1 to 2 weeks). The method should be considered as a potential "best option" in patients with recurrences and bilateral inguinofemoral hernias. PMID- 9408273 TI - Intraoperative ultrasonography for common bile duct exploration during laparoscopic cholecystectomy. AB - The "Endomedix Laparoscan Diagnostic Ultrasound" from the Medilas Ltd. and the "Leopard" and "Panther" type intraoperative ultrasound instruments from the Green Sound Ltd. have been used with success in sixty cases during laparoscopic cholecystectomy (LC) by the authors. Of these patients in four cases common bile duct (CBD) stones and in one case only sludge have been found. Of those with CBD stones, in one case four little stones have been observed despite of negative intraoperative cholangiography (IC) and in one case calculus in the retropancreatic CBD part was detected. On the basis of preoperative findings the CBD stones have not been expected. According to the authors the intraoperative ultrasound (IOUS) can be used with no exception in all cases eligible for LC, and also it can be used for the examination of neighboring organs (liver, pancreas, hepatoduodenal ligament). The method is extremely useful, performance quick, simple and informative as well as it can replace IC during LC. PMID- 9408274 TI - A case of fibrin sealant application for closing benign trachea-esophageal fistula (TEF). AB - Surgical treatment of upper airway-esophageal communications are frequently lead to failure. Having previous experience of thoracic administration of Tissucol fibrin glue authors attempted the local application in a case of TEF. A young lady suffering from myasthenia gravis required longstanding artificial ventilation. Not surprisingly a TEP developed in the area of the tracheostomy. There was no room for surgical repair of the TEF. Two administrations of rapid acting form of Tissucol was needed following local astringent therapy and enzymatic debridement to achieve a complete and permanent closure of a tracheobronchial sinus in a diameter of 5 mm. In addition of meticulous technique and of general supporting therapy special attention was paid to the followings: 1/healthy wound edges 2/ local infection control 3/dry environment 4/patient building up strategy. PMID- 9408275 TI - Effect of the DNA ploidity on survival in patients with carcinoma of papilla of Vater. AB - The authors investigated the possible prognostic factors for survival after pancreaticoduodenectomy for carcinoma of papilla of Vater. From 1984 to 1995, 8 patients underwent radical surgical intervention for tumor of papilla of Vater. The mean age of the patients was 58 years. Three of them were over 60 years. In one case Whipple procedure was performed, and pylorus-preserving pancreaticoduodenectomy was carried out in 7 patients. Using flow cytometry, the authors measured the nuclear DNA content of tumor cells. DNA ploidity status was evaluated from paraffin-embedded tumor tissues. Perioperative mortality occurred in one patient. Reoperation was performed on 2 patients, because of presence of anastomotic leakage. Survival was 50% at 1 year, 37.5% at 3 years, and 25% at 5 years. Tumor size (> 2 cm) was not negative prognostic factors for survival. The mean survival of patients with diploid cancer (n:6) was 17 months, and the mean survival of patients with aneuploid carcinoma (n:2) was 56 months. The proliferative index of the diploid carcinomas ranged from 3% to 11%. The proliferative index of the aneuploid tumors ranged from 17% to 28%. In conclusion, tumor size (> 2 cm), DNA ploidity status and proliferative index were not significantly negative prognostic factors for survival in patients with tumor of papilla of Vater. PMID- 9408276 TI - Keratoprosthesis. Implantation of artificial corneas. AB - Keratoprosthesis (implantation of artificial, plastic cornea) is indicated in severe cases with corneal leucoma (non-transparent, cicatrized cornea) in which keratoplasty (corneal transplantation) is not possible or has repeatedly failed. In the past 40 years we implanted 37 artificial corneas (7 Cardona type, 29 Konstantinov type, 1 Fjodorov type). The visual acuity increase was temporary (lasting from a few weeks to a few months) in 25 patients. The visual acuity was at least 0.2 three years following the implantation of keratoprosthesis in 12 patients. One patient had 1.0 vision 10 years after surgery. Our results indicate that the implantation of artificial corneal is still an "ultimum refugium", an operation that can be justified only in monocular patients, in eyes that cannot be and/or had unsuccessfully been operated on with repeated keratoplasties. The visual improvement is temporary, but in some cases may last for several years. Still this is the only procedure by which useful vision can be provided, for shorter or longer time intervals, for patients suffering from corneal blindness (nontransparent cornea in otherwise functioning eye) whose only eye cannot be treated with corneal transplantation. Keratoprosthesis with better biocompatibility, better fixation techniques, and wider visual fields have to be developed before the implantation of artificial cornea can be looked upon as a surgical procedure with which full optical rehabilitation can be achieved. PMID- 9408277 TI - Role of endotoxins and bile acids in the pathogenesis of septic circulatory shock. AB - It has long been known that the toxic effects of endotoxins under experimental conditions can be induced only when they are administered parenterally. However, in naturally occurring enteroendotoxemic diseases (e. g. septic and intestinal ischemic shocks) the endotoxins--which are produced by gram negative members of intestinal flora-, absorb from the intestinal tract to the blood circulation and can elicit pathological processes. It is an important distinction between natural and experimental endotoxin shock. If the common bile duct of rats were chronically cannulated a significant amount of perorally administered endotoxin was absorbed into the blood. This endotoxin shock can be prevented by bile acids. The physiological surfactants, the bile acids, are important facts in the defense of macroorganisms against endotoxins (physico-chemical defense). The production and passage of bile acids depend from the function of liver and the cholecystokinine (CCK) synthesis of small intestine wall. If the bile (bile acid) content of the intestinal canal decreases the endotoxin can translocate to the body and elicits toxic symptoms. So most important parts of defense against endotoxins in natural conditions are the CCK and bile acids. The consequence of damage of liver (place of bile acid synthesis) or small intestine (place of CCK synthesis) is the absorption of endotoxins. PMID- 9408278 TI - The current surgical treatment of primary malignant melanoma of the skin. AB - In our Department, between 1991 and 1996, 132 patients, there of 87 male-45 female, average age of 48.2 years underwent surgery because of stage 1 (T2) to stage 3 melanoma that was located on the skin. None of patients suffered from early or in situ melanoma. Our retrospective study was based on those 94 patients who had been followed up by Department or dermatology-oncology in our medical centre. Surgery is still the primary treatment for cutaneous malignant melanoma. Thin melanomas (up to 2 mm in thickness) can be excised with 2 cm margins. Whether this is also true for thicker melanomas is not known and the only way to obtain more knowledge is to participate in prospective randomised studies. Elective lymph node dissection is associated with significant morbidity which includes lymphedema, wound complications and paresthesias of the extremity. For this reason we use an alternative operative approach uses selective lymphadenectomy with identification of the sentinel node. PMID- 9408279 TI - Laparoscopic fenestration of symptomatic solid spleen cyst with harmonic scalpel instrument. AB - The treatment of two operated solid splenic cysts has been reported by authors. Laparoscopic cyst fenestration has been demonstrated to be a useful alternative method to open surgery. The aim of the authors was to analyse the use of Ultracision Harmonic Scalpel in two spleen preserving procedures. Cysts were located in the superior and the anterior-inferior pole of the spleen. Cyst wall not covered by spleen tissues was removed, drain was left in the abdomen. In the demonstrating spleen cyst operation the advantages of HS instrument was the clean operating field, correct coagulation of cyst wall, short hospital stay. Authors believe that this new technology will make it easier and more desirable for surgeons to fenestrate symptomatic spleen cysts. PMID- 9408280 TI - Laparoscopic hiatal reconstruction and use of harmonic scalpel instrument. AB - The method of laparoscopic repair of hiatal hernia is accepted in surgery. Usually associated with Nissen fundoplication which is the most commonly performed antireflux operation. Within a ten-year period authors have done 90 antireflux operations, 52 was laparoscopic procedures. (30 operations for GERD, 10 for hiatal hernia, 12 for the combination of both.) A patient was submitted to operation with large hiatal hernia. He was treated laparoscopic way with success and good results. The Ultracision Harmonic Scalpel instrument helped their operation in many ways, and provided correct bloodless preparation of cardiac region. The authors demonstrate our procedure and the use of the Harmonic Scalpel. Their patient were completely pleased with the results after discharge, they are still under regular control. PMID- 9408281 TI - Haemarthros induced articular cartilage degradation. AB - The statement that blood in the articular cavity is cause of cartilage degradation is widely accepted as an axiom. Although the causes of the different articular diseases were explained in numerous studies, none of them has clarified the pathomechanism of haemarthrosis. Our aims were: 1/ to give a morphological description of the blood induced changes in the cartilage, 2/ to verify that the haemarthros is the cause of the cartilage degradation. 10 white rabbits were used in our experimental model. Artificial haemarthros was produced in their left hind knees by intraarticular injection of their own blood. The right hind served as control. The rabbits were divided into to five groups based on the time of the haemarthros (22-50 days). Samples of the condylar cartilage were taken for light, polarization, transmission and scanning electron microscopy examinations. Signs of the disorganization of the matrix structure were showed by polarisation microscope and serious lesions were detected in the perichondrium by scanning electron microscope. Similarity have been suggested amongst the pathomechanism of haemarthrosis and other degenerative cartilage diseases (e. g.: osteoarthrosis, rheumatoid arthritis), so we made the same comparison. In many cases similar morphological changes were observed, as described by other authors in case of degenerative diseases. PMID- 9408282 TI - In vitro amylase release of preserved pancreas: a simple test to assess the viability of pancreatic allograft during preservation in the pigs. AB - To determine an in vitro marker of viability during pancreatic preservation, 12 pigs underwent total pancreas harvesting, and graft were stored in Euro-Collins or Belzer perfusion solution for up to 24 hours. Amylase concentration of the storage solution was analyzed in regular periods and tissue samples were taken for acridine-orange histochemical evaluation of viability in the same time. In vitro pancreatic amylase release (IU/g pancreas tissue) was calculated from the volume of solution and the weight of graft. A significant increase of amylase release was found in the course of preservation in both media. Comparing amylase release in different solutions we found significant difference between Euro Collins and Belzer media (4 hours: 6.45 IU/g vs. 2.2 IU/g, 8 hours: 11.5 vs. 3.58, 24 hours: 8.7 vs. 42.8, respectively). Comparison of amylase release with histochemical evaluation of viability showed strict correlation. We concluded that amylase release is a good marker for exocrine tissue destruction as well as viability of preserved pancreas. Our data confirms that Belzer solution is superior in pancreatic preservation. It is suggested that after adaptation into human model in vitro pancreatic amylase release could be a time- and cost-saving, useful method in predicting pancreatic transplant function prior graft implantation. PMID- 9408283 TI - Indications, new surgical technique and results of colon interposition or bypass in esophageal surgery. AB - Over a 5-year period, 29 patients with esophageal disease underwent colon interposition or bypass. The indication was cure of cancer in 11 patients, who underwent earlier a gastric resection. Other indications was benign stricture in 7 patients, bypass for unrespectable cancer in 6, having a caustic injury in 3 and after an esophageal perforation in 2. In 14 patients the left colon, in 15 the right colon was used. The colon was transected without dividing the mesentery other than just along its mesenteric border. This preserves additional blood supply from the marginal artery, also improves the function of the graft in transporting food. Anastomosis leakage occurred in 4 cases (13.7%). Graft necrosis occurred in 2 of 29 patients, one of whom alter underwent a successful second reconstruction. The 30 day operative mortality rate was 13.7%. A colon interposition provides good quality of deglution, and is the organ of choice for patients who require an esophageal substitute and are potential candidates for long survival, or when the stomach is unsuited for replacement or bypass. PMID- 9408285 TI - Posterior lumbar interbody fusion (PLIF) using the bony elements of the dorsal spinal segment. AB - Many different types of surgical technics for treating unstable degenerative spondylolisthesis of the lumbar spine has been reported. The most important part of the different procedures is to achieve a solid bony fusion between the two vertebrae. Authors report a new simple method of posterior lumbar interbody fusion in spondylolisthesis. The whole posterior segment of the mobile neural arch is removed and used for fusion. Corticospongiosus dowels and small bone chips are impacted into the disc space emptied totally before. Monosegmental transpedicular fixateur is required for stability. 33 patients with an average follow up of 20 months showed a clinical improvement of 88%. The radiologically proved fusion rate was 90%. Authors achieved good clinical outcome and high bony fusion rate with the one stage operation from posterior approach for treating lumbar spondylolisthesis. PMID- 9408284 TI - Experimental studies for the surgical correction and fixation of dorsal spine deformities. AB - One of the specific features of the scoliosis operations with a posterior approach is that both the correction of the deformity and then the maintaining of the corrected situation are carried out with the help of the implants. With the currently applied systems based on the CD principle it is still difficult to control the rotational component of the scoliotic curve. To complement the systems based on the CD principle, we have developed an implant family whose application makes the correction of the dorsal deformity generally simplier and derotation more effective. Our method is based on the application of such hooks which, linked to the longitudinal rods and hooked on both transverse processes of the instrumented vertebrae, transmit the concerted forces exerting their influence in the direction of the correction. Depending on their symmetrical relations, the hooks are capable of tilting in the frontal plane and derotating in the horizontal plane simultaneously, in the direction of our choice. The stability and applicability of the hooks were tested in implants into cadavers, and then the intimate relations of the implants were examined by means of radiological tests and dissection on instrumented specimens. On the basis of our results, the implants can already be used in clinical practice. PMID- 9408286 TI - Different techniques for creating oesophageal anastomoses. A historical review and personal experience. AB - Among the surgical techniques used to create a reliable oesophageal anastomosis, mention should be made of the handmade (in one or two layers, wire or Vicryl) and the different stapler anastomoses. 41 oesophageal anastomoses were performed by stapler technique between 4 March 1985 and 4 March 1991. The EEA stapler was used in 15, and the SPTU stapler in 26 patients. The average age was 56.8 years overall, 53.6 in the female (7 patients) and 57.4 years in the 34 male patients. Tumours in the middle and lower third of the oesophagus and on the cardia were the indications for resection in 30 instances. Total gastrectomy was performed in 9 patients and oesophageal resection for peptic stricture in 2 cases. Replacement with stomach was carried out after oesophageal resection (17 patients), and with Roux-loop in 24 cases. The EEA anastomoses were not covered by a hand-made layer of interrupted sutures as is compulsory in the case of the SPTU gun. The intraoperative complication rate was 12.2%--two severe complications with the SPTU and 3 mild ones with the EEA (2 cases) and SPTU (1 case) machines. The postoperative complication rate was 17%--the severe ones with the SPTU gun. The only fatal anastomosis insufficiency was observed in this group. 3 of the 41 patients died--a mortality rate of 7.3%--but only one of them was due to technical failure in the SPTU group: 2.4%. CONSEQUENCES: Both the intra- and postoperative complications were more severe with the SPTU technique. The early postoperative complications are closely related to the intraoperative ones. Mortality due to technical failure was only observed in the SPTU group. The EEA stapler gun is superior in every respect to the SPTU sewing-machine. The future belongs to the even more sophisticated bent and modifiable devices. These were used in our Department Between 1992 and 1997, with practically no morbidity and no mortality. Although they are the most expensive of all the possibilities, the low morbidity and mortality rates pay off from the aspects of the short hospital stay and the savings in human life. PMID- 9408287 TI - Primary resection with antegrade colonic irrigation and peritoneal lavage versus subtotal colectomy in the management of obstructed left colon cancer. AB - Between 1980 and 1996 122 patients with acutely obstructed resectable carcinomas of the colon and rectum were treated in our hospital. Ninety-four has undergone one stage operation of immediate resection and primary anastomosis without proximal colostomy. Intraoperative colonic irrigation was performed in 34 patients, subtotal/total colectomy in 30 patients, right hemicolectomy in 30 patients. There were 2.3 and 1 operative deaths, respectively. The average hospital stay for the survivors was 19.8 days in the primary resection and anastomosis group, and 41.0 days in staged operation group. Concerning the possibility of multiple lesions the authors recommend subtotal/total colectomy except for tumours localized in the sigma where they apply total colectomy only when the proximal part of the colon is necrotized or filled with solid stool. PMID- 9408288 TI - Trend from laparotomy to laparoscopy in the case of treatment of ectopic pregnancy. AB - Ectopic pregnancy is a very hazardous, life threatening complication of the women. Diagnosis and treatment previously were very conservative. When not typical symptoms were found, only waiting or observing was the main task of hospitalisation and only the beginning of typical signs came laparotomy. Nowadays, with the introduction of vaginal ultrasound, color doppler or power imaging techniques and sensitive blood HCG analysis greatly reduced hospitalisation period. The need for the change from laparotomy to laparoscopic treatment in surgical operations became everyday practice. We analyzed the data in our department for the operation procedures of ectopic pregnancies from 1990. PMID- 9408289 TI - Minimally invasive laparoscopic surgery on experimental animal models. AB - Our goal was to find a very good model for gynecological laparoscopic operations. The main purpose of the operations was to perform the same types of laparoscopic operations as we use daily in our clinical practice at the Department of Obstetrics and Gynecology. UMSD. The uterus of female dogs seemed to be ideal for this work. In every experiment we carried out 10 different interventions on identical part of different dog uterus. The operations were performed at identical time period of day (a.m.). The weight of the female dogs were similar. The anaesthesia was also performed the same way in every case. The main purpose of the operations was to find some significant changes or differences between the 10 different surgical techniques. The hystological investigation were carried out with normal microscope and electron microscope. PMID- 9408290 TI - The values of detection of free radical mediated reactions in patients. AB - The peroxidative processes and individual antioxidant protection were measured in patients with different cardiovascular diseases. We concluded that monitoring of this system we were able to detect not only the actual changes of lipid peroxidation and antioxidant defence mechanisms, but additionally the therapeutic efficacy of the treatment. PMID- 9408291 TI - Results of contact beta-irradiation of choroidal melanomas. AB - Between the end of 1986 and the end of October 1996 81 patients suffering from choroidal melanoma were irradiated at the Department of Ophthalmology University Medical School of Debrecen. The irradiations were performed with Ruthenium-106 beta ray emitting scleral plaques (ophthalmic applicators). The radioactive plaques were implanted surgically, fixed by episcleral sutures onto the sclera covering the base of the tumour, and removed by another operation when sufficient time for irradiation had elapsed. For the treatment of tumours in the vicinity of the optic nerve special plaques were used with a notch for the nerve. Precise localisation of the tumours was established by intraoperative ultrasonography and transillumination. The mean follow up period was 3.5 years after the irradiation. Tumour regression was recorded by fluorescein angiography, ultrasonography and fundus photography. In 75 cases (92.5%) the treatment was successful. Additional laser photocoagulation was performed after irradiation in 26 cases. The irradiation by Ruthenium-106 scleral plaques of choroidal melanomas proved to be an effective treatment in cases of small and medium sized choroidal melanomas in our practice and the results are similar to those of the largest radiotherapy centers of Europe. PMID- 9408292 TI - Integrin expression on normal and neoplastic human breast epithelium. AB - Integrin adhesion receptor expression of different benign and malignant breast tumours was examined by means of immunohistochemical techniques. A panel of seven different anti-alpha and two different anti-beta subunit antibodies was used. Normal breast epithelium displayed a well characterized and constant pattern of integrin expression consisting of strong alpha 1,2,3,6 and alpha v, and a relatively weaker beta 1 and beta 3 staining. No staining for alpha 4 or alpha 5 could be detected on the epithelial cells. Benign fibroadenomas did not show changes in their receptor expression compared to normal tissues. In the cases of different types of breast cancer, there was a significant downregulation of all subunits. The staining pattern was distinct if there could a basement membrane like structure be detected around the invading tumour nodules. When laminin and collagen type IV surrounded the tumour cells, those cells in contact with the extracellular matrix components still displayed strong positivity for the integrin subunits. Other cells inside the tumour cell nests or not surrounded by basement membrane did not express integrins. The positively staining cells might be more differentiated owing to the effect the basement membrane. Myoepithelial labeling of the integrin expressing cells gave negative results. The observed integrin expression heterogeneity renders the histologic picture difficult to interpret with regard to clinical behavior of the tumour. PMID- 9408294 TI - Late complications after pharyngogastrostomy. AB - A case of severe stricture and a case of tracheogastric fistula after laryngopharyngo-esophagectomy and pharyngogastrostomy for cervical esophageal cancer are described. Stricture is often seen but tracheogastric fistula is a rare complication, however, both are devastating conditions. According to the literature, the survival rate is poor in both cases. The surgical management demands several principles. Recurrent or metastatic cancer must be ruled out. The patients' general condition and nutritional status must be optimized. Pulmonary infection must be cleared. The surgical management of the stricture was a free jejunal transfer after failed attempts of several dilation procedures. The treatment of tracheogastric fistula was suturing the stomach and covering the trachea with a pedicled left sternocleidomastoideus flap. The survival of the patient treated with free jejunal interposition exceeds 24 month. Unfortunately, the patient with tracheogastric fistula, treated with interpositioned sternocleidomastoideus muscular flap, lived two weeks after this operation. The surgical managements described in this report may provide palliation or definitive treatment for these devastating complications. PMID- 9408293 TI - Anaesthesiological indications and contraindications of minimally invasive surgery. AB - Spreading of laparoscopic techniques caused changes in anaesthesiological contraindications. In the first period laparoscopy was contraindicated in ischemic heart disease (IHD). Early mobilisation and short postoperative period are positive goals, IHD was taken out of contraindications. Present study compares changes in circulatory, blood gas and acid-base balance values during laparoscopic cholecystectomy (LC) in groups of patients ASA I-II. and ASA III. with IHD. There were 30 patients in group ASA I-II, 30 patients with IHD in category of ASA III. investigated during LC. Fifteen patients of both groups went under Propofol-Fentanyl (TIVA) anaesthesia, others were on Propofol-Fentanyl-N2O (IVA) protocol. All of them got also Atracurium. Pulse rate, mean arterial pressure, O2 saturation and end tidal CO2, blood gases and acid-base state were recorded before induction, after CO2 insufflation, after desufflation, 1 and 3 hours postoperatively. After CO2 insufflation there was a moderate tachycardia in both ASA III. groups (74/min-->88/min). In all groups pCO2 increased (40-->48 mmHg) but normalised till the 3rd postoperative hours (42 mmHg). Ventricular extrasystoles appeared in 3 ASA III. patients in IVA group. Three high risk patients had serious metabolic acidosis postoperatively. Present time the ischaemic heart disease does not contraindicates laparoscopic interventions. TIVA with Propofol is better choice because of its favourable effects on circulation and acid-base balance. Using N2O caused higher grade of intestinal distension. The cardio-respiratory, blood gas parameters and acid-base balance have to be monitorised in perioperative period of laparoscopic surgery. PMID- 9408295 TI - Changing techniques and indications for lamellar keratoplasty. AB - In recent decades, the number of lamellar keratoplasties (LKs) being performed worldwide has been gradually decreasing. However, new technical procedures have been developed and microsurgeons must consider modified indications for lamellar grafting. In the period 1946-1995, in the Department of Ophthalmology University Medical School of Debrecen, Hungary 3889 consecutive keratoplasties were performed of which 608 were LKs. In the period 1946-1960 special surgical material (biosutures) was used without operating microscopes and the leading indication for LK was corneal scar (leukomas). Between 1961 and 1979 with the development of microsurgical procedures (10/0 nylon sutures, operating microscope), the LK technique became more advanced. In the period 1979-1995 precision lamellar grafting could be performed with the widely available improved modern techniques. Changing indications for LK were noted in the second period and this tendency has subsequently continued. Currently the main indications for LK are "immediate" keratoplasty after chemical burns, and sclerokeratoplasty against recurrent pterygia. Their observations lead the authors to conclude that the possible reasons for the changing indications of LK are the new technical procedures, the increasing knowledge, the better immunological tests and the improved postoperative therapy. PMID- 9408296 TI - Experience acquired by applying gentamicin-sponge. AB - Chronic suppuration is a reverting problem in surgical practice. The concerned region either has insufficient circulation due to basic disease, or systemic antibiotics takes no effect due to inflammatory barrier developed meanwhile. For this reason Authors initiated systematic use of gentamicin sponge (Garamycin, Schering-Plough, USA) two years ago. During this period we implanted gentamicin sponge in 40 cases. After primary intervention in 85% of treated patients we observed complete recovery, or in case of ulcus cruris we reached proper condition for skin-transplantation. Three patients needed further incision and in two cases amputation of diabetic foot could not be avoided. Our results support the effectivity of gentamicin sponge in treating local inflammatory processes supplementing and increasing efficiency of surgical procedures. PMID- 9408297 TI - The pathological plica in the knee. AB - Fourteen patients were evaluated who had an arthroscopic removal of the symptomatic mediopatellar plica of the knee in this retrospective study. The main complaint of the patients was the pain at the medial side of the knee. Before the operation only in 3 cases was the diagnosis the pathological plica. The arthroscopy was performed in a routine way. The plica was consider pathological, and removed, when it was thickened. There was grade I or II chondropathy in the knee on the medial femoral condyle and the patella respectively, caused by the plica. Good to excellent result were obtained in 93% of the knee with or without chondromalacia. PMID- 9408298 TI - Relevance of cytokine production to infected pancreatic necrosis. AB - The purpose of this study was to evaluate the role of cytokines in septic conditions following acute pancreatitis and to elaborate a new strategy in the treatment. Increased TNF and IL-6 serum levels were found in 30% of the patients (n = 40), while the IL-6 level was elevated in all of them. There was a positive correlation between the serum IL-6 and sICAM-1 levels. The in vitro TNF and IL-6 producing capacities were initially higher in the study group, but decreased on subsequent days, especially in fatal cases (n = 3). Administration of pentoxifylline [PTX] (400 mg/day) to septic patients following necrotizing pancreatitis resulted in TNF and IL-6 production similar to that observed in control donors. The level of sICAM-1 also decreased following PTX therapy. These results suggest that cytokines produced by activated leucocytes are important in the pathogenesis of infected pancreatic necrosis, and their inhibition might be of therapeutic advantage. PMID- 9408299 TI - TGF-beta 1 and IL-6--new aspects in pancreas regeneration? AB - Plasma levels of interleukin-6 (IL-6) and transforming growth factor-beta (TGF beta 1) were studied during cholecystokinin octapeptide (CCK-8)-induced regeneration after pancreas resection in rats. The weight of the pancreas and the DNA and protein contents increased significantly. The serum levels of TGF-beta 1 and IL-6 were increased significantly on days 7 and 14. There was no significant change in serum amylase levels. These findings indicate that cytokines such as TGF-beta 1 and IL-6 may play a role in the pathomechanism of pancreas regeneration. PMID- 9408300 TI - Why the internal mammary artery is an ideal graft for myocardial revascularisation? (an experimental model with omentoplasty). AB - Ten dogs underwent coronary artery bypass grafting through a right thoracotomy. Free internal mammary artery grafts (IMA) was used for aortocoronary bypass to right coronary artery (RCA) without temporary cardiopulmonary bypass. In upper median laparotomy approach, mobilization and lengthening of the omentum was performed through the retrosternal part of the diaphragm. An epiploic muff was effectuated surrounding the IMA graft. After three months the animals were sacrificed and the grafts folded by omentum were used for microscopical study. The histological evaluation has shown that the omentum has formed a new adventitia around the graft. In the new-formed adventitia, newly formed vessels can be found and these are grouped around the adventitia-media border zone, very rarely penetrating into the external layer of the media. This graft-omentoplasty offer a good experimental model which facilitates to investigate in vivo the blood supply needed by free IMA grafts through the adventitia in experimental conditions. The biological mechanism of angiogenesis in graft wall can be investigate as well. PMID- 9408301 TI - Late epigastric incisional hernias following laparoscopic cholecystectomy. AB - By the introduction of laparoscopic cholecystectomy a new "gold standard" procedure became a routinely performed operation in the field of biliary tract surgery. Thus, the incision related early and late complications are thought to diminish, especially the formation of incisional hernias. Five patients had been referred to our department suffering from chronic incisional hernias following laparoscopic cholecystectomy. All of the hernias were located to the site of the epigastric trocar. The contents of the hernias proved to be omentum. The documentation's of the laparoscopic cholecystectomies revealed the extraction of thick walled gallbladders that contain large stones, and the wounds through which the extraction was performed had not been closed. Taking into consideration the fact of the "Chimney Effect" caused by the desufflation of the pneumoperitoneum at the end of the laparoscopic operation, bowel or omentum can easily escape through the relatively large wound formed during the extraction of the gallbladder, resulting in the formation of incisional hernias. This can be avoided by the complete desufflation and the prompt closure of the wound. PMID- 9408302 TI - Inflammatory mediators and surgical trauma regarding laparoscopic access: free radical mediated reactions. AB - In this prospective study the free radical mediated reactions, the changes of endogenous antioxidant defense mechanism and activation of leukocytes were measured from the blood of patients undergoing elective cholecystectomy because of symptomatic cholecystolithiasis. The patients were randomised into two groups. Group one contained 21 patients treated by open cholecystectomy(OC). Group two consisted of 21 patients treated by laparoscopic cholecystectomy (LC). Both groups had similar patient characteristics. Patients with acute cholecystitis, pancreatitis, choledocholithiasis or other disease were excluded. Values from patients in both groups were compared. The measured biochemical parameters are the following: malondialdehyde (MDA) as a marker of the free radical induced lipid peroxidations, reduced and oxidised glutathione (GSH-GSSG), as endogenous scavengers as well as markers of oxidative stress and myeloperoxidase activity (MPO) of leukocytes. The results showed significantly lower values of postoperatively measured MDA, GSH-GSSG, and MPO activity of leukocytes in patients with laparoscopic cholecystectomy, indicating a lesser stress response and tissue trauma in this group of patients. The results correspond to the favourable results of most other trials evaluating clinical aspect of LC. PMID- 9408303 TI - Acute arthroscopy. AB - The role and significance of acute arthroscopy have been evaluated in the treatment of knee joint injuries on the basis of findings during 59 arthroscopic operations which were conducted within two weeks after the accident. The injuries developed isolated in more than half of the cases (65%) whereas they appeared in a combination of two or more in 26% and in 9%, respectively. Injuries requiring operation were found in 91.5%, most of which were ruptures of the ACL (33 cases) and menisci (23 cases). In the case of ACL rupture, in the acute phase on sportsmen and physical workers primary arthroscopically assisted transligamental replacement was performed with patellar graft while in the case of proximal rupture of the ACL reinsertion and augmentation were carried out with semitendinosus tendon. The ruptures of dislocated eminentia were refixed in each case. In the case of the rupture of meniscus the refixation of the meniscus of resection of the ruptured part was attempted. By means of acute arthroscopy the lesion of the intraarticular structures or that of their combinations can be exactly diagnosed. Depending on the findings of arthroscopy the injuries can be treated immediately or operated on at a later time, thus preventing the joint from further deterioration. PMID- 9408304 TI - Therapeutic value of continuous passive motion after anterior cruciate replacement. AB - The aim of this study is the evaluation of the therapeutic value of continuous passive motion after ACL replacement. After 41 ACL reconstruction in 13 cases only active motion, in 28 cases active motion and CPM were used postoperatively. Stability, the range of motion and complications were evaluated at the time of discharge, 3 and 6 months after the operation. The flexion-extension in the CPM group was significantly greater at the time of discharge, but this difference was practically eliminated in 6 months after the operation. There were not any other significant difference between the two groups. The CPM gives only a little advantage in the rehabilitation after the ACL replacement. PMID- 9408305 TI - Prognosis of untreated liver metastasis from rectal cancer. AB - Majority of studies based of treatment of liver metastases from colorectal cancer compare their results to historical controls that include patients having untreated secondary liver tumours. The aim of this study was to show the natural history of patients with liver metastasis from rectal cancer. Data of 303 patients underwent laparotomy between 1984 and 1992 were reviewed. 47 of 57 patients who had liver metastasis at the time of surgery fulfilled the criteria of the study. The mean survival time was 8.5 (1-27) months for all patients, 11 months for patients with solitary hepatic tumour (n = 13) and 7.5 months for those with multiple tumours (n = 34). Patients who had liver and other distant metastases simultaneously (n = 9) survived a shorter time than those with hepatic secondaries only (4 and 9 months respectively). The mean survival time for patients in whom the primary tumour was resected (n = 17) was 11 months contrary to 7 months for those who underwent colostomy or exploratory laparotomy, but comparison of survival curves didn't revealed significant difference between the two groups. CONCLUSIONS: The incidence of liver metastases from rectal cancer at the time of laparotomy was 19%. The prognosis was very poor in case of synchronous secondary tumours. Palliative resection of the primary tumour is recommended for selected patients only to control local symptoms. PMID- 9408306 TI - Our preliminary results in application of Stoppa technique for recurrent groin hernia. AB - In 1994 we have started the Stoppa technique which was only used for the recurrent groin hernia repair, when Mersilene mesh was implanted in preperitoneal position from a lower midline incision. From March, 1994 to March, 1996 seventeen Stoppa procedures were performed in our department (14 men, 3 women, mean age of 66 years). Type of hernias: 13 unilaterale, 3 bilaterale recurrent groin hernias and 1 primary bilaterale groin hernia combined with a lower midline incisional hernia. Operations were performed by 7 surgeons. The mean time of the surgery was 85 min. (35-165). The mean postoperative stay in the hospital was 9 days (7-19). The Stoppa technique could be performed in all patients. Serious early postoperative complications didn't occur. The two late recurrences were observed amongst the first five patients. All implanted mesh were well tolerated by the patients. Author believe, this procedure has some advantages: the site of the incision is located in a new place from where the dissection is easier than in the previous incision, the stich fistulas from the previous operation can be avoided, furthermore this method could be applicated for combined hernias located in the lower part of abdomen and it is cheaper than the laparoscopic hernioplasty. PMID- 9408307 TI - Interdisciplinary treatment of the malignant lymphoma of the thyroid. AB - 720 patients with malignant tumor of the thyroid were treated at our department between 1950 and 1996. Histological examination revealed 15 cases (2%) of malignant lymphoma (14 female, 1 male mean age: 69.8 year). In 8 cases Hashimoto thyroiditis, in 1 case chronic lymphocytic thyroiditis was verified beyond the lymphoma. According to Ann Arbor staging, 9 patients were stage I E, 3 II E, 1 III E, and 2 IV E. Radical surgical treatment was performed in 7, palliative in also 7, and only biopsy in one case. Irradiation was administered in 9, chemotherapy in 8 patients. In 5 out of the 7 palliative operated patients tracheostomy was performed. 10 patients were lost. The therapy of the non-Hodgkin lymphoma of the thyroid is debated in the literature. There are some authors, who question the role of surgery. Our experience, based on the longer survival of the patients, who underwent radical operation, support those, who consider the possible surgical treatment as the initial part of the multimodal therapy. PMID- 9408308 TI - Creation of axillo-axillar bypass with bidirectional anastomoses. AB - Scholz and co-workers reported such type of anastomosis applied in femoro-crural bypass procedures which leads blood flow well into both directions. In vascular surgery practice of the author there were several occasions where this modified anastomosis was favorably applicable. An old man was operated up in this manner whose left common and internal carotid artery and the starting part of the subclavian artery was occluded that caused steal syndrome. Axillo-axillar bypass was done with end to side anastomoses. After the operation blood pressure increased to 155/75 mmHg on both arms the steal phenomena ended. Bidirectional anastomoses make flow from obtuse angle to rectangular. PMID- 9408309 TI - About streamlined brachial dialysis shunts. AB - The authors modified the technics of fistula forming. In such cases where to create a connection on the forearm is impossible one possible modality is to make it between major vessels on the arm. Streamlined connection was made in seven patients between basilic vein and brachial artery in three cases, between cephalic vein and brachial artery in four. There were two men and five women. The age interval was between 58 and 78 years. The longest observation period is 30 months. Until this time the fistulas were suitable for dialysis and had abundant yield. Because of the number of cases and the observation period has not yet make possible to come to final conclusion their goal was to present technical solution first of all. Their expectation is that with making them streamlined they will manage to create good perfusing and long lasting fistulas. PMID- 9408310 TI - The influence of Roux-Y versus Longmire-Gutgemann reconstruction on the composition of pancreatic juice. AB - Both methods of reconstruction after gastrectomy lead more or less to an insufficiency of pancreas. Therefore investigations on rats should further clarify which defects are obvious after both operation methods. Wistar rats were divided into 3 groups. In 2 groups a gastrectomy was performed while one was reconstructed according to the method of Roux-Y, the other was treated according to the method of Longmire-Gutgemann. The first group was a sham operated control group. 3 months after this operation pancreatic juice was collected over the time of 6 hours. Volume and protein content were determined as well as a differentiation of the proteins by means of the 2D electrophoresis which separates the molecules according to isoelectric focus and the molecular weight. The results show a significant increase of the volume of pancreatic juice after both operations. Whereas the protein content is also altered the number of proteins is significantly decreased. Especially proteins with an alkaline isoelectric focus are significantly diminished. The molecular weight of the proteins is also changed. Low molecular protein fragments which were not observed in the sham operated group are increased especially in the Roux-Y group. This means that the production of enzymes is changed after both operations. The pH optimum as well as the viability of the protein enzymes is shifted. Since the changes are more pronounced after Roux-Y operation signs of pancreatic insufficiency should be expected more frequently after this operation. PMID- 9408311 TI - Is the presence of distant metastasis associated with c-myc amplification in gastric cancer? AB - The expression of the c-myc oncogenes has already been reported in human gastric carcinoma. Overexpression can be the consequence of oncogene amplification and often correlates with different prognostic factors. Authors investigated the value of c-myc oncogene amplification in 23 patients (9 male, 14 female, aged 28 85 yrs) with gastric cancer and its correlation to the following clinical and histopathological parameters: grade, TNM stage, Lauren's type, localisation and severity of disease. DNA was isolated from formalin-fixed, paraffin embedded tissue for quantitative dot-blot hybridisation. Amplified c-myc was found in 6 out of 23 cases. Its values ranged from 2.12 up to 18.2 (average 9.1). Significant association was found between the presence of c-myc amplification and distant metastasis (corr. coeff.: 0.5623, p < 0.01). High scores of the other parameters also correlated with c-myc, albeit not significantly. The result of cluster analysis, based on the similarity of the parameter values for the individual patients proved that the age was the decisive factor in creating two groups. The distribution of patients into these groups did not seem to coincide with the presence of c-myc amplification or distant metastasis, inspite of the proved correlation between them. PMID- 9408312 TI - Excimer laser photorefractive keratectomy with different ablation zones. AB - In this study we would like to introduce the excimer laser, and to demonstrate our results and complications by using different ablation zones during photorefractive keratectomy (PRK) in the correction of myopia and astigmatismus. In 1996 we performed photorefractive keratectomy on 100 myopic eyes of 52 patients (28 females, 24 males). Mean age was 26.21 years (ranged from 19 to 54 years). The preoperative refraction ranged from -1.0 D to -18.0 Diopters. The diameter of the ablation zones were between 5 and 6.5 mm. We evaluated the results and the complications of the surgeries of 100 eyes which were performed with Schwind keratom F excimer laser. After 2 days, 1 week, 1 month, 3 months, and 6 months postoperatively we tested the best uncorrected and corrected visual acuities, and performed intraocular pressure measurement, slit lamp examination as well as corneal topography. The postoperative refractions were between +/- 0.5 to +/- 1.0 Diopters. After six months postoperatively the slit lamp examination showed that 80% of the patients had no corneal haze while 20% had stage I (Hanna) corneal haze. The smaller the diameter of the ablation zone was, the more pronounced the corneal haze and the night-glare were. The photorefractive excimer laser keratectomy is judged to be a safe method, although it might have some side effects. The different ablation zones of this treatment means an important modification, that not only allows the method to meet the individual requirements, but reduces the chance of the complications as well. Based on the authors' experiences PRK for moderate myopia with large diameter ablation zones appears more predictable than than with smaller ablation zone diameters. PMID- 9408313 TI - The role of different tumor markers in the early diagnosis and prognosis of pancreatic carcinoma and chronic pancreatitis. AB - The examination of tumor markers in the diagnosis and in the evaluation of progression of tumors has got an increasing significance. The serum level changements of three tumor markers (CEA, CA 19-9, CA 125) were examined before and after the operation in 94 patients operated for pancreatic carcinoma (PC) and chronic pancreatitis (CP) between March 1994 and December 1996 at the 2nd Dept. of Surgery of Debrecen Medical University. From the patients 62 were operated for carcinoma, in 19 cases the tumor was resectable, 43 patients had palliative operation. In 32 patients ductal decompression was performed because of CP. The authors evaluate the serum level changements of the three tumor markers examined in three groups of patients before and after the operation. In conclusion CA 19-9 is the most sensitive marker of PC, the sensitivity was 77.4%, the specificity was 87.5%. CEA and CA 125 are not as sensitive markers of PC as CA 19-9, while CEA and CA 125 serum levels are both increased in half of the patients with chronic pancreatitis. PMID- 9408314 TI - Second-look surgery (SLO) in the management of carcinoma of the ovary. AB - There is a permanent controversy on the clinical relevance of second-look laparotomies. In the first approach the majority of clinical experts evaluated this procedure experimental. Our results support the idea of clinical relevance of this procedure as patients with SLOs through the early detection of recurrence really profited from the procedure. PMID- 9408315 TI - Minimal invasive surgery. AB - The advent of laparoscopic cholecystectoma has established a new chapter in surgical treatment with an impact on the various surgical specialties leading to significant changes in surgical practice. The essential attribute of the new approach is the reduction of the trauma of access. Minimal access surgery (MAS) underlies all the benefits of the new approach including the accelerated recovery. Few other developments in surgery have excited so much interest among surgeons and the medical equipment industries alike, and progress during the past 7 years has been impressive though at times lacking direction and in some instances, scientific backing. PMID- 9408316 TI - The replacement of the esophagus by musculocutaneous flaps. AB - If there is no other possibility for the replacement of the whole esophagus, the antethoracal neo-esophagus from musculocutaneous flaps gives the best result compared with the skin tube reconstruction. Two successful cases are discussed. PMID- 9408317 TI - 10 years of cooperation with the Department of Experimental Surgery, University Medical School of Debrecen: scientific outcome and impact on clinical practice. AB - From 1986-1996 several series of experiments were done in cooperation with the Department of Experimental Surgery, University Medical School of Debrecen. Very interesting scientific findings have been achieved in the fields of urethral surgery, continent urinary diversion, alternatives to dialysis, laparoscopy and bladder stimulation. Besides representing a high scientific output nearly all experiments had direct impact on our clinical practice. PMID- 9408318 TI - Inflammatory mediators and surgical trauma regarding laparoscopic access: acute phase response. AB - Numerous studies have tried to compare different aspects of patient response to laparoscopic cholecystectomy (LC) versus open cholecystectomy (OC). Our study focused on the acute phase response in order to clarify the better patient recovery following LC. Sixteen patients scheduled for elective cholecystectomy were equally allocated into groups of OC and LC. Blood samples were collected before and for four days after the procedures. Levels of interleukin 6 (IL-6) and C-reactive protein (CRP) were determined by ELISA technique. IL-6 value increased ten fold on the first postoperative day (52.8 pg/ml) in the OC group with a return to baseline value by the fourth postoperative day. In contrast a moderate increase postoperatively (12.1 pg/ml) with a fast normalisation of IL-6 value by the second postoperative day was noted in LC group. A similar pattern was observed in the CRP level. CRP peaked to 84.1 mg/l by the second follow-up day after OC, while only an insignificant rise to 52.7 mg/l was registered in the LC group with again a faster return to baseline value. The marked contrast between the two groups with regard to IL-6 and CRP changes clearly underlines a diminished acute phase response following LC, which verifies LC as probably a less traumatic procedure. PMID- 9408319 TI - Lymphadenectomy in gastrointestinal surgery for malignancy. AB - In a randomised study 25 patients with gastrointestinal surgery combined with extended lymphadenectomy (three field lymphadenectomy in case of esophageal cancer, D2 lymphadenectomy in case of gastric cancer) has been compared to the same number of patients with limited lymphadenectomy (D1). The operation time and the need for blood transfusion has increased in the extended lymphadenectomy group. The complication rate was more than doubled in the extended lymphadenectomy group, due to fluid or lymph collection, lymphatic edema, and infection. The mapping and staging was superior in extended lymphadenectomy group, but increased morbidity and mortality has been found in this group. However the favourable effect of extended lymphadenectomy on survival needs further long-term studies and proofs. PMID- 9408320 TI - Transverse hepatectomy in surgical treatment of gallbladder carcinoma. AB - In the surgical treatment of gallbladder cancer not only the traditional resections (right trisegmentectomy, lobectomy), but newer, parenchyma sparing multisegmentectomies, resection of 4B, 5, 6, that is transverse resection is also justified, as it is shown by demonstrating the 2 first cases in Hungary with transverse hepatectomy. World-wide there is a tendency for more aggressive surgical treatment in case of gallbladder cancer. For this reason we introduced transverse hepatectomy in our surgical practice, and in this paper we demonstrate the technical details of the new procedure. The goal of the transverse resection is to remove the tumour with an adequate margin of resection. The remaining hepatic parenchyma allows good quality of life for the patients till recurrence develops. PMID- 9408321 TI - Experiences with duodenum preserving pancreatectomy. AB - According to the principle of surgery for chronic pancreatitis the preservation of pylorus, duodenum or distal part of common bile duct gives the benefit of more physiological intervention. 2 patients with duodenum preserving pancreatectomy are presented. The operation was carried out for chronic pancreatitis. Both patients had jaundice and needed T drainage. Both patients suffered from very severe malnutrition with cachectic condition adding severe pain. None of them proved to be malignant by the frozen section. Previous diabetes, severe chronic inflammation of the whole pancreas, destruction of the pancreatic ductal system and cysts helped the decision-making for ablation of pancreas with preservation of duodenum which seems organ saving procedure. In comparison with the Whipple operation the duodenum-preserving pancreatectomy spares the patient a gastrectomy, a duodenectomy and a resection of distal common bile duct. PMID- 9408322 TI - Preoperative work up and therapeutic schedule for patients with duplex tumors. AB - Authors analyze the cases of duplex and multiplex tumors among the more than two thousand operated upon patients a year at the Surgical Department of Uzsoki Hospital, Budapest, Hungary. The study enrolls a two years period evaluating the medical history of our patients in hope of a more sophisticated and effective diagnostic and therapeutic regimen. Our experience gained by analyzing data of our patients with synchron tumors shows that the patient's life expectancy is much better if performing a radical multivisceral tumor extirpation at the same sitting even if keeping in mind the higher perioperative risk. In accordance to the above mentioned concept we would like to stress the utmost importance of the interdisciplinary cooperation. PMID- 9408323 TI - Impaired T cell functions preceding lymphoproliferative disorders in mice neonatally tolerized to transplantation antigens. AB - In A/J (H-2a) (A) mice, the neonatal i.v. injection of (B10 x A)F1 spleen cells (SC) induces partial transplantation tolerance (TT) to C57BL/10ScSn (H-2b) (B10) skin allografts, chronic host-versus-graft disease (HVGD) and lethal lymphoproliferative disorders (LPD). They produce anti-T-cell autoantibodies (ATA), and the proliferative responses of their SC to the T cell mitogen Con A are decreased. We found that, similar to ATA, the hyporeactivity of T cells developed earlier (at 1-2 weeks of age) than splenomegaly. The proportions of both CD4+ and CD8+ T cells were not reduced in the spleens of tolerized mice without manifest LPD. The supernatants (SN) of Con A-stimulated tolerized SC contained no, or only small amounts of interleukin-2 (IL-2). Thus, in the tolerized mice, ATA and T cell deficiency preceded the development of LPD. ATA and the decreased amount of the T cell growth factor IL-2 might play a role in the defective T cell activation. PMID- 9408325 TI - Use of harmonic scalpel at laparoscopic cardiomyotomy. A new method. AB - The laparoscopic cardiomyotomy with anterior fundoplication (Heller-Dor procedure) is accepted for treatment of esophageal achalasia. The crucial point of the procedure is proper myotomy and avoid perforation of the esophagus. Hook cautery is widely accepted to dissect the oesophageal muscle. We'd like to demonstrate our experiences with a new device--Ultrasonically Activated Harmonic Scalpel-, witch was used at our operation for achalasia to make the cardiomyotomy. Between December 1993 and December 1996, 11 patients with esophageal achalasia underwent laparoscopic Heller's operation with Dor's antireflux procedures. In one patient we applied the Ultrasonically Activated Harmonic Scalpel (HS) to make the cardiomyotomy. The use of HS and results were evaluated. Application of the Harmonic Scalpel is effective for cardiomyotomy. It can be used more safe than electrocoagulation, because it cause less thermic lesion. It's easy to use at laparoscopic way. A perforation of the esophagus didn't occur. There was no intra-, or postoperative complications. After the operation the patient is free of complains. After our first operation, we have found, that Ultrasonically Activated Scalpel can be applied with safe and good results for the cardiomyotomy at laparoscopic operations for esophageal achalasia. PMID- 9408324 TI - Effects of antiendothelin treatment on the early hemodynamic changes in hyperdynamic endotoxemia. AB - We have performed a series of experiments to study the effects of a newly developed antisense homology box-derived endothelin (ET) antagonist peptide (ETR P1/fl) on the early hemodynamic changes in a hyperdynamic endotoxemic dog model. Mean arterial pressure (MAP), cardiac output (CO) and myocardial contractility (MC) were measured in closed-chest animals. Plasma levels of ET-1,2 were determined by radioimmunassay. A hyperdynamic circulatory response was elicited with a 2-hour infusion of 5.3 micrograms/kg of E. coli endotoxin (ETX). Control and ETX-treated animals received an infusion of ETR-P1/fl (0.1 mg/kg) i.v. ETX treatment decreased MAP and MC, increased initially CO, and a long lasting elevation in the plasma ET level was observed. In ETX-treated animals the administration of ETR-P1/fl significantly prolonged the increase in CO and inhibited the depression of MC. Our results suggest that treatment with the ET antagonist ETR-P1/fl may be advantageous in the early phase of endotoxemia. PMID- 9408326 TI - Use of harmonic scalpel for division of short gastric vessels at laparoscopic Nissen fundoplication. A new method. AB - The essential of laparoscopic Nissen fundoplication is creating a loose and tension free wrap requiring mobilisation of the gastric fundus. Division of the short gastric vessels (SGV) is a standard component of that procedure which requires considerable part of the operation time, and despite of a careful dissection sometimes significant blood loss can be occurred. The Ultracision Harmonic Scalpel (HS) makes the process quicker safer and can cause less intraoperative complication. This study compares our original method of vessel control (at 10 cases, Group 1), to the Ultracision Harmonic Scalpel (at 10 cases, Group 2) Times for SGV division, estimated blood loss and intraoperative and postoperative complications were evaluated. In the favour for Group 2 a significant reduction was reached in the time required for division of SGV to mobilize the fundus. As the most common postoperative complication, the dysphagia concerns we found significant difference in the two groups and dysphagia was mild at Group 2. Application of HS provide safer, easier and faster division of SGV resulting significant savings of time and less prone to thermic trauma and recommended at LNF operations to avoid potential intra- and postoperative complications. PMID- 9408327 TI - Resection of giant schwannoma by combined surgical exposure. AB - Contrary to the past experience the giant Schwannoma with symptoms of canalis vertebralis compression has been removed by combined surgical exposure in one sitting. Laminectomy, decompression of the canalis vertebralis and immediately subsequent extracanalicular resection of the tumour by retroperitoneal approach was performed in one sitting by two surgical teams. The advantages of the combined surgical exposure: 1. The affliction of the patients caused by the operation significantly decreased. 2. The expenses of the treatment, nursing and hotel decreased as well. 3. The surgical team of different specialists remove the tumour together in both approaches. PMID- 9408328 TI - Diagnostic-staging laparoscopy. AB - Recently instead of the old fashioned, traditional explorative laparotomy the newer up-to-date procedure, the diagnostic-staging laparoscopy has been generally used in the everyday surgical practice of the Authors. Evaluating 32 diagnostic and staging laparoscopy Authors draw the attention to the importance of this procedure which has high diagnostic value, and reduces the complication rate. Biopsy, cytology, frozen section can be done. Authors carries out diagnostic and staging laparoscopy in cases of esophageal, liver, stomach and pancreas tumours just immediately before surgery. PMID- 9408329 TI - Sutureless anastomosis in the surgery of the gastrointestinal tract. AB - The idea of the sutureless anastomosis is not a new one, but only the introduction of the VALTRAC-BAR instrument made the wide clinical application possible. The authors have applied this instrument in the case of 32 patients in a two years period. It was used in intestinal procedures in 28, in gastric resection in 4 occasions, respectively. In one case spontaneously recovered stercoral fistula was observed. According to our experiences, the instrument can be used advantageously in preparing anastomoses in the gastrointestinal tract. PMID- 9408330 TI - The blood supporting of nipple-areolar (NAC) complex performing for mammaplasties. AB - In case of mammaplasties the blood support of the nipple-areolar complex is one of the most important. The authors give a comprehensive clinical summary of the operating technics which are suitable to correct mastoptosis and breast hypertrophy (or macromastia). After mentioning the anatomy, blood and nerve supply of breast, following the classifying of mastoptosis and breast hypertrophy. Summarizing of the historical development of operating methods, which resulting in modern solutions, and adding Hungarian-related data. The widely used mastopexies and reduction mammaplasties will be analyzed, with demonstration of transposition of NAC using different kind of dermo-glandular flaps, e.g.: Skoog-, Pitanguy-, McKissock-, Strombeck-, Robbins-, Regnault- and other's method. After discussing the possible postoperative complications, authors underline the necessity of detailed analysis and exact planning in varied clinical cases for achieving wanted functional and aesthetic result. Attaching importance to have more perfect operating techniques. PMID- 9408331 TI - Level of sensitivity of pain in patients with obesity. AB - Our aim was to find out the presence of differences in pain sensitivity level depending on body weight. 206 healthy persons at the age of 18 to 84 were the objects of the research with the help of the special instrument which action was based on the principle of dosage pressure by a needle on the forearm skin and by registration of indecis pressure extents when the patients felt no pain, when they felt slight pain and when they felt violent pain. The patients with normal body weight didn't feel pain when the pressure of a needle was 28.53 g; persons with surplus body weight and fatness of the 1 degree-46.5 g and persons with fatness of the 2-3 degrees-61.55 g. Slight pain was registered when the pressure was 40.5 g; 57.6 g and 72.9 g conformably. And the violent pain-76.5 g; 91.3 g and 116.2 g. Thus the patients with fatness have higher pain sensitivity threshold then people of other categories, so they feel less pain. The older a person is the higher pain sensitivity threshold is. PMID- 9408332 TI - Internal carotid stent implantation with angioscopic control. AB - Stent implantation is a method, which is being used more and more often, mainly in the field of peripheral arteries, but coronary stent implantation is also well known. The authors apply this procedure for patients, who besides the carotid bifurcation stenosis also suffer from the internal carotid stenosis in a longer section of the artery. The indication are as follows: 1 in the case of internal carotid stenosis in a longer section we assure the flow with the help of carotid thrombendarterectomy and slove the run-off with balloon catheter dilatation and stent implantation. 2. in the case of a stenosis in a short section we use stent implantation to avoid dissection of the intima. With each intervention we perform the carotid bifurcation thrombendarterectomy. The authors made the first intervention 15 months ago, since then we have performed 17 stent implantations. One of the 17 patients developed a temporary stroke (TIA). This is a new method, further cases are needed for long-term experiences. Intervention under eye control can be carried out with more confidence. As regards to the post-operative medicinal treatment, the administration of thrombocyte aggregation-blockers or heparinoid preparation (e.g.: PPS-SP 54) after the stent implantation has become a routine therapy. Summarized the foregoing, these cases constitute about 10-15% of all carotid operations, therefore the authors would like to make it clear, that this is not a routine method, but a possibility, strictly respecting the above-mentioned indications. PMID- 9408333 TI - Endogenous intoxication syndrome when acute process in abdominal cavity. AB - In the time of simultaneous study of the components of blood of sick patient and animals with acute processes in abdominal cavity it was revealed that in short periods there is increase in value indeces of endogenous intoxication syndrome. The independent index is offered for studying the endotoxicosis of whole blood. It is recommended to use for general and particular characteristics of dynamics during the pathological process. PMID- 9408334 TI - Surgical treatment of pancreatic head and periampullary tumors. AB - The aim of this study was the comparison of the postoperative results of standard Whipple pancreatoduodenectomy (WP), pylorus preserving pancreatoduodenectomy (PPPD) and palliative bypass operation performed for treatment of pancreatic head and periampullary tumors. In the period from Jan. 1992 to 1996 106 patients had tumors located in the head of pancreas and 21 patients had periampullary tumors. The diagnosis was established by ERCP, transabdominal ultrasonography and computer tomography. We assessed the morbidity, mortality, prognostical data of the surgery of pancreatic head and periampullary tumors. Tumor markers such as CEA, CA 19-9 and CA 125 were also studied. The operability rate was 26% in case of pancreatic head tumors and 69% in peri ampullary tumors. The mortality rate was 6%. Postoperative complications were in 23 patients(18.1%). There was no significant difference between the survival of WP and PPPD group, but we found much better survival in patients with periampullary tumor. After palliative operation the survival rate was 6.1 months in case of pancreatic head carcinoma and 11 months in case of periampullary tumors. Our data provided many evidences about the advantage of PPPD in the patients with malignant periampullary and pancreas head tumors and the long-term results and quality of life is much better after PPPD. PMID- 9408335 TI - Improvement of swallowing ability in advanced oesophageal cancer. AB - Without different types of palliation the patients with inoperable oesophageal cancer have a poor quality of life, rapid weight loss which leads to death. The aim of palliation is the complete relief of dysphagia. Our modified procedure is a simplified way of a well known method described by Tytgat in 1986. The prosthesis is positioned under continuous visual control using only local anaesthesia. This method is safe and not expensive. During the last three years 73 consecutive patients were treated with palliative fiberoscopic intubation with Tygon prosthesis. 46 patients had esophageal carcinoma, 19 had gastric, 8 had pulmonary carcinoma obstructing the gullet. Among them 11 patients had bronchoesophageal fistula. The early complications were perforations (7) bleeding (2), and later complications: food impaction (5) tumor overgrowth (5) and tube migration (2). The mortality was 2%. All patients have received antibiotic prevention. Although the improvement of life quality is more important than extension of life, for many patients survival will be prolonged due to improved nutrition as a result of treatment. This study summaries our experience with this technique and analyzes the problems and complications encountered in our patients. PMID- 9408336 TI - Morphology of cystic renal lesions. Lectin and immuno-histochemical study. AB - Renal cystic disease include heritable, developmental and acquired disorders. Morphological features were extensively studied mainly in cases of autosomal dominant polycystic and experimentally induced cystic disorders. We report the immunohistochemical (cytokeratin, epithelial membrane antigen, vimentin, Tamm Horsfall protein, proliferating cell nuclear antigen) and lectin-binding (soybean agglutinin, Dolichos biflorus agglutinin) profile of cystic kidneys from 9 surgically removed and 21 autopsy cases. The primary renal diseases displayed great diversity. Beside polycystic kidney diseases we studied cysts associated to renal neoplasm, hemodialysis, nephrosis syndrome and chronic transplant rejection. Cystic epithelium demonstrated positive reactions with distal tubular markers (epithelial membrane antigen, cytokeratin) or collecting duct (soybean agglutinin, Dolichos biflorus agglutinin) and Henle loop markers (Tamm-Horsfall protein) but the latter in lesser extent. The large number of the vimentin positive cases are suggestive to dedifferentiation or cellular regeneration. The former might be underlined by the diffuse cytoplasmic or basolateral membrane staining of the epithelial membrane antigen in some cystic epithelial cells. Not the cystic epithelium but rather the neighbouring non-dilated tubular cells and interstitial cells presented proliferative activity which was most intense in areas where vimentin and variable nephron segment markers in the same tissue were expressed. Positive reaction of the type IV basement membrane collagen and the rate of the inflammation failed to show similar connection. This finding suggests the importance of the inflammatory cells in the development and/or expansion of the cysts. PMID- 9408337 TI - Prospective randomized trial comparing Shouldice and Bassini-Kirschner operation technique in primary inguinal hernia repair. AB - The authors conducted a prospective randomized trial to compare the Shouldice and the Bassini-Kirschner technique between April, 1994 and December, 1995. During this period 129 adult patients, mean age 54 (17-87) years underwent operation on primary inguinal hernias in their department. 63 Shouldice and 66 Bassini Kirschner operations were performed by 17 surgeons. The duration of surgery, the technique of anesthesia, the perioperative complications, the duration of postoperative care, then one year after the operation the recurrence rate and the patient's subjective complains were investigated. 85 patients of 129 were examined one year after the operation in spite of that all the patients were invited for control. Both operations gave almost the same results in the perioperative period and nearly identical recurrence rate (4.44% in Shouldice and 5.0% in Bassini-Kirschner group). The patient's subjective complains were also very similar one year after the operation. The authors could not find significant difference in the results of the two types of surgery although they were well experienced with the Bassini-Kirschner operation and just starting to practice the Shouldice procedure. These facts suggest that having more experience with the Shouldice operation makes available the extreme low recurrence rate published by several authors. PMID- 9408338 TI - Jejunal nutrition. AB - Contrary to the past experience of forced parenteral nutrition nowaday's the enteral [jejunal] nutrition enjoys priority. It is not questionable, that well adjusted and controlled application of fluid, ion, fat, carbon hydrate, amino acid promoted convalescence. The experiences of the Authors supports that enteral nutrition through technically proper outperformed jejunostomy does not increase complication rate and beside well controlled food administration provides the physiologic stimules of food, the method is relatively easy and cost effective. For this reason the Authors initiated jejunostomy at the end of larger interventions such as Akyama procedure, total gastrectomy, multivisceral interventions, pancreatectomy, operations for massive gastrointestinal bleeding and finally reoperations with extreme negative N-balance and with the chance of inability of oral feeding for several days. PMID- 9408339 TI - Changing concept and modern techniques in cataract surgery. AB - Until the middle of the 80-ies the routine method of cataract surgery was intracapsular extraction (ICCE). Approximately 10 years ago the so called extracapsular cataract extraction (ECCE) started to become more and more widely used. After extracapsular cataract extraction had come into general use, it became possible to implant the IOL behind the iris, into the left in place capsular bag of the original lens. We usually perform extracapsular cataract extraction with posterior chamber lens (PCL) implantation as a routine procedure. In the last few years phacoemulsification started to gain acceptance. During this operation one breaks the nucleus of the lens into pieces with the help of ultrasound, and this way it is possible to remove both the nucleus and the cortex of the lens through a small wound. The first step in learning phacoemulsification is to be able to create a perfectly round hole on the anterior capsule (capsulorhexis). Besides using various manual techniques we introduced diathermal capsulotomy for capsulorhexis. This latter procedure is becoming more and more popular as it is safe, easy to perform, and gives excellent result. PMID- 9408340 TI - The effects of bisaramil and antiarrhythmics on free radical generation of isolated neutrophil granulocytes. AB - The aim of this study was to investigate the effect of bisaramil--an antiarrhythmic drug under clinical trials--on free radical generation of isolated polymorph neutrophil granulocytes (PMN) and to compare its activity with well known antiarrhythmics. PMNs were isolated from healthy beagle dogs, and superoxide radical generation was induced by phorbol-myristate-acetate. Free radical generation capacity of stimulated PMNs were measured. Bisaramil exerted a concentration dependent inhibitory effect on stimulated free radical generation. At the investigated concentration range of the antiarrhythmics only propafenon, mexiletine and diltiazem showed similar activity as bisaramil, but clear concentration dependency could not be seen in any of the cases. According to the results of this study inhibition of stimulated free radical production by isolated PMNs can not be closely related merely to either membrane stabilizing or Ca-antagonistic activity of drugs. In vitro inhibitory action of bisaramil on free radical generation indicates a possible cardioprotective effect existing independently of its antiarrhythmic one. This observation may be important in outlining the range of clinical indications of bisaramil as it may also be useful in the treatment of reperfusion injury. PMID- 9408341 TI - Monitoring of plasma total antioxidant status in different diseases. AB - The pathological increase of oxygen free radical generation has already been recognised in more than one hundred diseases. To gain information about the consequences of oxidative stress the investigation of plasma antioxidants seems to be plausible. In our study we used a new kit (RANDOX, England) for measurement of total antioxidant status (TAS) to determine whether it has diagnostic value in comparison with our earlier results of measuring other parameters of oxidative stress in the following diseases: i./In the group of patients with ischemic heart disease (n = 19) the TAS elevated from 1.08 +/- 0.13 to 1.16 +/- 0.11 mM after 2 weeks of cardioprotective drug administration showing the beneficial effect of drug treatment. ii./In the group of patients with essential hypertension (n = 47) its values were below the normal range (1.11 +/- 0.15 mM) at the time of the first investigation and increased gradually following antihypertensive treatment. iii./The changes of TAS values of patients who underwent open (n = 21) or laparoscopic (n = 21) cholecystectomy indicated the less surgical trauma following laparoscopic procedures. Our results suggest that determination of TAS is a valuable and reproducible method to detect the actual antioxidant status in patients. PMID- 9408342 TI - The effects of glucocorticoids and a glucocorticoid antagonist (RU 38486) on experimental acute pancreatitis in rat. AB - The effects of glucocorticoids on acute pancreatitis are a matter of dispute. In animal experiments, dexamethasone and hydrocortisone significantly decreased the serum amylase activities 8 hours after the induction of pancreatitis. In the dexamethasone treated group, the serum IL-6 level was significantly decreased at 4 and 8 hours, while in the hydrocortisone treated group, all the IL-6 values were significantly diminished vs. the control group. As compared to the control, a glucocorticoid antagonist (RU 38486) did not influence the serum amylase activity, but significantly increased the serum IL-6 level. These results suggest that glucocorticoids may play a role in the control of pancreatitis caused by inhibition of cytokine production. PMID- 9408343 TI - Experimental investigation of preservation injury in animal kidneys after reperfusion with Euro-Collins. AB - The authors evaluated the pathomorphologic alterations of removed and reperfused dog kidneys by means of light and electronmicroscopic examination. In each sample the following reversible signs were found: Hypereosinophilia (HE), Hydropic dystrophy (HD), Nuclear polymorphism (NP), Epithelial desquamation (ED), Brush border lesion (BBL), Single cell necrosis (SCN), Total tubular epithel necrosis (TTEN), Interstitial edema (IE), Perivascular edema (PE). The irreversible signs were: Basement membrane rupture (BMR), Cellular infiltration (CI), Glomerular mesangial matrix expansion (GME) and vascular lesions (VL). The most severe and mostly irreversible alterations occur in the 54-72 hours after harvesting. The authors emphasize the significance of basement membrane rupture, because the impossibility of tubular epithelial regeneration, the cellular infiltration due to its fibrogenic effect, glomerular lesion because it makes decrease the glomerular filtration rate, proceeding juxtaglomerular cell proliferation and hypertension through renin-angiotensin mechanism and vascular lesions causing renovascular hypertension and tubulopathy. The authors believe that reperfusion injury is very important factor in kidney allograft survival. Its mechanism is similar to the normal necrosis pathway, but the timing is delayed. Further investigations are needed to understand what specific alterations may occurred under blood circulation in the host to reveal more exact cause of primary graft failure after transplantation. PMID- 9408344 TI - Intraepithelial neoplasms of the pancreas. AB - Intraepithelial neoplasms of the pancreas have been recently described and relatively rare, but these lesions represent a distinct, however pathologically heterogeneous entity with shared clinical features. We analyzed the clinicopathologic and cytogenetic characteristics of eight patients with intraepithelial neoplasms. Based on our hypothesis that tumor ploidy pattern correlates with ploidy, synthetic (S) phase and proliferative fractions of each neoplasm by flow cytometry. Analysis of the nuclear DNA content of pancreatic intraepithelial neoplasms in this study supports our hypothesis. Each neoplasm demonstrated diploid stemline, with a low S-phase fraction. The diploid DNA pattern and the low proliferative activity are consistent with the nonaggressive behavior of the tumors and, in part, explain the favorable prognosis of intraepithelial neoplasms. Intraepithelial neoplasms of the pancreas constitute a rare, but surgically curable, localized disease with a good prognosis following radical resection. The most valuable tool in the diagnosis of these preinvasive lesions is ERCP combined with brush cytology. This study supports the potential value of flow cytometric DNA analysis in determining outcome, as the diploid DNA pattern and low S-phase fractions correlate with the nonaggressive biological behavior. PMID- 9408345 TI - The part of MR cholangiography in biliary surgery. AB - The magnetic resonance cholangiography (MRC) is a new none invasive examination procedure which demonstrates well the extrahepatic ducts. The examination can reveal all the anatomical or pathological anomalies of the biliary tracts. The authors performed this examination on 12 patients between 1st December 1996 and 31st December 1996. In 4 patients there were no pathological changes. Common bile duct stone was found in 1 patient. Sclerosis of the Vater papilla in 1 case. Mirizzi syndrome in 1 case. Carcinoma of the common bile duct in 3 cases. In 2 cases inflammation of the head of the pancreas caused compression on the bile duct. The patients were all operated. The operative and the MRC diagnosis corresponded. Similar examination procedure has not yet been mentioned in Hungary. PMID- 9408346 TI - The Lichtenstein open tension-free hernioplasty. AB - In the department of general surgery, in Szolnok MAV General Hospital the authors performed 51 open tension-free herniorrhaphies by Lichtenstein method between 1st July 1995 and 31st December 1996. According to this method the rings are closed anteriorly with Dacron mesh. The method is simple and rapid, less painful than any other technique. Apart from two minor complications, our patients recovered without any problem. The method helps to reduce the period of hospital treatment as well as that of sick-leave. PMID- 9408348 TI - Surgical treatment of gastric cancer: a retrospective study with special reference to epidemiology. AB - The present retrospective study gives the analysis of the epidemiological data of 1474 patients with gastric cancer, who were operated on at the 1st Department of Surgery of the University Medical School of Debrecen in the course of 35 years. In spite of the changes in diagnostics and therapeutic management it was impossible to raise, during this period, the rate of resectability: in the first 10 years it was 56%, and 55% in the last five years. There was no improvement in the proportion of curative and palliative resection, either: 50/6 vs 49/6. The number of gastrectomies increased to 22.5% from 17%, in a quarter of the cases, in the last two periods, extended multivisceral interventions were also performed. In the location of resectable carcinomas the number of those spreading over more than one third increased from 2% to 7%, and similar growth was encountered in the rate of diffuse-type carcinomas. The postoperative surgical complications decreased to 18% from 27%, and early postoperative death from 19.6 to 10.1%. In the last two periods 46% of the patients were admitted in advanced stages of tumor (UICC III B or IV), with half a year's or longer histories. Exclusively surgical treatment is not effective in these cases. PMID- 9408347 TI - Distribution of calcitonin-containing parafollicular cells of the thyroid in patients with chronic lymphocytic thyroiditis: a clinical, pathological and immunohistochemical study. AB - The C-cell hyperplasia of the thyroid gland is recognized as precursor to medullary carcinoma, particularly in multiple endocrine neoplasias, however it can be associated with hypercalcemic states and follicular tumours as well. The authors analysed 46 of their cases with Hashimoto's and/or lymphocytic thyroiditis from the immunohistological point of view with the aim of determining to what extent is C-cell hyperplasia associated with these pathological pictures. C-cell hyperplasia was demonstrated on sections of intraoperative preparations with the immunoperoxidase method with anticalcitonin MoAb. Moderate, focal C-cell hyperplasia was found in 17.4% (8 patients), extensive focal hyperplasia similarly in 17.4% (8 patients). Diffuse C-cell hyperplasia occurred in 1 case (2.2%). These results suggest the possibility that in the case of diffuse or more pronounced focal hyperplasia the serum calcitonin concentration of these patients is also elevated. Further clinical and pathological studies are needed to find out whether there is some pathogenetic relationship between chronic lymphocytic thyroiditis and C-cell hyperplasia or whether the finding is coincidental, and to establish what degree of C-cell hyperplasia is associated with elevated serum calcitonin levels. This relationship has been unknown until the present time, whereas it can be of great clinical significance, in part because of the changes in serum calcitonin levels, in part for screening out the MEN IIa type and working out the operating strategy. PMID- 9408350 TI - The prognostic value of CA-125 in epithelial ovarian cancer patients during and after chemotherapy. AB - In the past ten years the investigations on CA-125 levels of epithelial ovarian cancer patients have raised several questions both in the screening and in the follow-up. The predictive value of the test during the postoperative chemotherapy and the follow-up period was the topic in the series of examinations. Our findings are as follows. The decrease of the CA-125 level predicts a long tumour free survival. The permanent high level of CA-125 is a signal of a recurrence within few month after stopping therapy. In case of increase of the CA-125 level an early recurrence can be expected. PMID- 9408349 TI - Lymphonodal response to sensitization with 2,4-dinitrochlorobenzene (DNCB) in laboratory rats. AB - White laboratory rats were used for the experiments. Their popliteal lymph nodes were investigated after sensitization with DNCB (2,4-dinitrochlorobenzene) and i.c. injection of 3-H-thymidine and Patent Blue Violet at several intervals following the challenge with the hapten DNCB. Changes of weight of lymph nodes, further uptake of tritiated thymidine by the cells were investigated by auto radiography, while the content of 3-H-DNA indicating an increased DNA metabolism due to blastic transformation of the small lymphocytes was determined by a liquid scintillation method. The authors found a significantly increased weight of lymph nodes and a higher percentage of labelled cells following antigenic stimulus of i.c. injected DNCB compared to non-sensitized lymph nodes of the control rats. PMID- 9408351 TI - Pancreatic pseudocysts associated with chronic pancreatitis--early and late results of 1367 operations. AB - The authors hereby review the data of 1367 operations for pancreatic pseudocysts. The surgical procedures of choice in particular pancreatic pathologies are analysed in the light of early morbidity and mortality, as well as long term follow-up results. The best operations for pancreatic pseudocysts have been the internal drainage procedures, which resolve the pathological alterations without the necessity of pancreatic resection. The treatment of chronic pancreatitis may require combined surgical procedure, such as cysto-Wirsungo-gastrostomy. The pancreatic resections performed for the treatment of small pseudocysts in the pancreatic head have been superseded by the less invasive blunt, forced cysto duodenostomies, representing better results secondary to the smaller perioperative risk for the patient. The cyst-to-stomach and cyst-to-duodenum internal drainage techniques are just as effective, but with shorter operation time, than the Roux-en-Y cysto-jejunostomies. PMID- 9408352 TI - Application of minimally invasive surgery in Mayer-Rokitansky-Kuster-Hauser syndrome. AB - With the advancement of endoscopic techniques more and more invasive procedures can be replaced by less harmful interventions. Successful laparoscopic treatment of vaginal agenesis in Mayer-Rokitansky-Kuster-Hauser syndrome is described in four patients. PMID- 9408353 TI - Results with collagen fleece coated with fibrin glue (TachoComb). A macroscopical and histological experimental study. AB - Diffuse bleeding from parenchymatous organs at conventional surgery is eliminated with the usual methods coagulation tamponade or styches. We performed experimental series at 9 dogs. After resection of spleen, liver, pancreas and kidney, the bleeding surface was covered by collagen fleece coated with fibrin glue (TachoComb). Postoperatively 7 days, 10 days, 14 days and 28 days we made a relaparotomy. Then the results were analyzed macroscopically and microscopically. In the abdominal cavity neither significant quantity of blood nor greater adhesions were detected. At all cases the fibrin glue was found on place were it was put before. Histologically a perfect wound healing experienced. The fibrin glue (TachoComb) using at diffuse parenchymatous organs' bleeding give a very good results when the wound area is at least 1 cm beyond the immediate wound margin and the fibrin glue is applied onto the wound and pressed on it for 4-5 minutes. PMID- 9408354 TI - Plasma levels of TNF and IL-6 following induction of acute pancreatitis and pentoxifylline treatment in rats. AB - Activation of cytokine cascade is a decisive factor in determining the pathobiology of different inflammatory processes including acute pancreatitis. The purposes of this study were to determine the TNF and IL-6 levels after the induction of acute necrotizing pancreatitis, and to establish the effects of pentoxifylline on the cytokine production and the severity of pancreatitis. Acute necrotizing pancreatitis was induced by the retrograde injection of 200 microliters taurocholic acid into the pancreatic duct in male Wistar rats. TNF was titrated in a bioassay on cell line WEHI clone 164. IL-6 was measured via its proliferative action on the IL-6 dependent mouse hybridoma cell line B-9. Seven mg/kg pentoxifylline was administered intraperitoneally at the time of operation and/or 24 hours later. Rats were sacrificed, 48 or 72 hours after the operation. The TNF bioassay revealed high levels of TNF (36.6 +/- 6.0 U/ml) in the control group whereas levels decreased to zero in the pentoxifylline-treated group. The IL-6 bioassay likewise demonstrated high levels of IL-6 in the control group and markedly decreased levels in the pentoxifylline treated group (7083 +/- 2844 pg/ml, 6463 +/- 1307 pg/ml vs. 137.5 +/- 85.5 pg/ml, respectively, p < 0.05). The high mortality observed in the control group (43%) was sharply decreased by pentoxifylline administration to 11%. The data suggest that pentoxifylline is capable of modifying the cytokine production after 48 hours of induction of acute pancreatitis. PMID- 9408355 TI - Comparison between continuous brain tissue measurement and cerebrovenous measurement of pO2, pCO2 and pH in a porcine intracranial pressure model. AB - Simultaneous oxygen measurements in brain tissue (p(ti)O2) and hemoglobin saturation measurement in cerebrovenous blood in patients after severe head injury have shown different results regarding the comparability of the findings in respect to CPP and ICP. This is contrast to theoretical expectations. The aim of this study was to compare continuous ptiO2 measurement with oxygen partial pressure measurement in sagittal sinus (pO2cv) during simultaneously performance in an animal intracranial pressure model. For continuous measurement we used a newly available multisensor probe. We placed a Paratrend 7 probe (BSL, High Wycombe, UK) in the left frontoparietal white matter and measured ptiO2, pCO2 (ptiCO2) pH (pHti) and temperature (t(ti)) while simultaneously measuring these parameters (pcvO2, pcvCO2, pHcv, tcv) in the sagittal sinus in 9 pigs under general anaesthesia. A fogarty balloon catheter was placed supracerebellar infratentorial and inflated stepwise in order to increase ICP. The baseline levels of pO2ti, pCO2ti und pHti in the non-injured brain tissue showed a more extended heterogeneity compared to the findings in cerebrovenous blood. Both, pO2ti and pO2cv were significant correlated to the CPP decrease. In both measurement compartments pCO2 was inverse correlated to the course of CPP and seems the course of pH mainly to determine. p(ti)O2 as well as p(cv)O2 showed a close correlation to the CPP course and have proven to be qualified to indicate metabolic information about the relation of cerebral blood flow and metabolic cerebral demands. The measurement of CO2 tension in both measurement compartments shows a distinct heterogeneity of the absolute values and the results are only weak correlated to CPP. Metabolic influence on this parameter could not be revealed in the used experimental approach. PMID- 9408356 TI - Details of teaching, learning and training in laparoscopic surgery. AB - The authors discuss the specific details of teaching, learning and training in laparoscopy in 6 main sections, on the basis of their experience gained through basic and advanced courses held for approximately 480 participants at the courses organised at the Department of Experimental Surgery of the University Medical School of Debrecen, between 1989 and 1996. The 6 section cover the following: 1. Why these courses are needed? 2. Who should participate in them? 3. When courses should be held (continuously for beginners, at an appointed time for advanced participants, and chances for training should be provided at any time, according to demand) 4. What should be taught? 5. Where teaching and training should take place (surgical learning/training centres) and 6. What methods of teaching should be used. PMID- 9408357 TI - Protection of the renal artery in nephron-sparing surgery. I. Pathomorphological study. AB - It is necessary to clamp the renal artery for some time in renal surgery to preserve the organ. Not only the renal parenchyma but also the wall of the clamped renal artery may be damaged due to known ischaemic-reperfusion causes. Regional venous renal hypothermia induced by intracellular solutions (Sacks II, Euro-Collins) could provide protection against damage of the vessel wall caused by reperfusion or clamping in our experiments, in cases of clamping for 30 minutes, probably because it also ensured the cooling of the surface of the vessel wall. Similar vasoprotective effect could be achieved by systematic antioxidant treatment (MTDQ-DS) and regional renal hypothermia given in combination. Harmful effects of 45-minute clamping could be best avoided by applying antioxidant treatment alone. While there were no significant morphological changes in the renal arteries the persistent endothelial damage may trigger further complications like renal ischaemic condition. PMID- 9408358 TI - Protection of the renal artery in nephron-sparing surgery. II. Arterial contractility investigations. AB - It has been well known that reperfusion following ischaemia may cause functional and structural damage to not only the organ involved but also the blood vessels supplying that organ. As in organ-sparing renal surgery it is inevitable to clamp the renal artery for some time, it is expected that reperfusion, following the removing of clamping, causes structural changes in the vessel wall which may result in a decrease in arterial function. In our model experiments on animals, the left renal arteries were atraumatically clamped for 30, 45 and 60 minutes. Simultaneously with clamping, perfusion regional renal venous cooling was applied to some of the animals, together with nephrotomy. In some cases cooling was performed in combination with antioxidant treatment. On the 3rd postoperative day renal arteries from both sides were removed, the right, intact ones serving as control. Noradrenaline dose effect curves characterizing vessel contractility were determined to demonstrate functional changes. It was established that cooling the renal artery for only 30 minutes was enough to rule out the damage due to ischaemia-reperfusion. If clamping lasted for 45 minutes, venous cooling of the kidney in combination with antioxidant treatment was necessary to spare arterial function. Clamping for 60 minutes resulted in irreversible/permanent decrease in contractility even if hypothermia and antioxidant treatment were given simultaneously. PMID- 9408360 TI - Modifications of and special operating tricks in the classical technique of surgery against strabismus. AB - Even nowadays the idea of most of the operations against strabismus is based on myectomy and reposition of the rectus muscles. At the Department of Ophthalmology of the University Medical School of Debrecen we use mainly these two methods of operation. At our Department we performed 1605 operations against strabismus during the last 10 years. 1269 of our patients had convergent, 336 had divergent strabismus. The majority of the operations were performed on patients less than 6 years of age, under general anesthesia. The outcome of the operation is successful if it results in a situation, in which torque affecting the eyeball stabilizes it in the normal position. The advantage of the combined surgical procedure is that the former muscular balance remains intact, and by relatively small intervention good results can be achieved. In the rectus muscles of squint patients anatomical and histopathological changes could be found, especially if they did not receive pleoptic treatment preoperatively. Depending on the direction of the strabismus one of the muscles is thicker, while the other is thinner, due to hyper- and hypofunction. Our surgical experience of many years showed that by modifying the classical technique of the operations with some fine technical tricks it was possible to increase the success rate. In our film we presented the right way of incising and handling the subconjunctival connective tissue (Tenon capsule) thus making it possible to lay it back to its original place at the end of the operation. This way the chance of postoperative scarring is reduced. We demonstrated how to test the contractility of the muscles by using muscle-hooks, that replaces the forceps test. In case of myectomy we demonstrated the tricks of how to handle the easiest and the finest way. In case of retroposition we can achieve perfect reconstruction and wound healing by using the shown simple technique. We also take much care of suturing the conjunctiva during operations performed on infants that is important because of their undisciplined behavior. PMID- 9408359 TI - Intraoperative monitoring of the motor pathway using transtracheal stimulation of the cervical spine in dogs. AB - Although SEP monitoring of the spinal cord has been a well established method recently, not an ultimate, perfectly developed technique for monitoring of the motor system is known so far, particularly, because of the disturbing effect of narcotic drugs and relaxants on the motor evoked potentials. In this study the upper part of the spinal cord was stimulated in 14 anesthetized and relaxed dogs with a cathode attached to the intratracheal tube and an anode fixed to the cervical spinous processes. Single and serial stimuli were applied. Recordings were obtained from the exposed right femoral nerve and quadriceps muscle. Averaging was necessary when using serial stimulations. Responses were consequent and reproducible during regular anesthesia. The origin of the different responses in the spinal cord is discussed. The method seems to be appropriate for intraoperative monitoring of the thoracolumbar spine. PMID- 9408361 TI - Thoracic and lumbar sympathectomy with the application of ROMICRO mini laparotomy set. AB - The authors present the use of ROMICRO set (originally developed for mini laparotomy) for thoracic and lumbar sympathectomy. They review 3 thoracic and 2 lumbar sympathectomy operations that had good effects and uneventful recoveries. The advantage of the method is that no special instruments are needed and it can be done as a conventional operation. PMID- 9408362 TI - Intrastromal corneal ring, a new refractive surgical technique to decrease myopia. Experimental and clinical results. AB - From among the wide range of keratorefractive operations, the authors applied the intrastromal corneal ring (ICR) technique to treat myopia. The results of operations performed on dogs' eyes were evaluated using slit lamp investigations, cornea topography and histological investigations. In the possession of favourable results, this kind of operation was successfully performed on three patients suffering from expressed unilateral myopia, too. The desired optical results (10.0 dioptres) proved to be permanent 3-10 months after surgery and were demonstrated by the patients' visual acuity as well as cornea topographic examinations. The surrounding of the PMMA ring situated intrastromally and the substance of the cornea remained calm and clear, respectively. The authors think that the ICR technique can successfully be applied in the treatment of myopia similarly to refractive surgical interventions such as radial keratotomy (RK) and excimer laser photorefractive keratectomy (PRK), which are widely used methods these days. PMID- 9408363 TI - Multimodality treatment of pancreatic pseudoaneurisms. AB - Vascular lesions of pancreatitis manifest in the form of haemorrhage into the pseudocyst (PC), the development of pseudoaneurisms (PA) or splenic lesions. Between 1987 and 1996 31 patients were found to develop vascular lesions either in the form of haemorrhage into a PC (12) or PA (19). Diagnosis of pancreatic PA was established preoperatively in 8 cases only. Gastrointestinal (GI) bleeding manifested in 12 patients, but only in 6 of them was the pancreatic origin of the bleeding considered. All patients were operated. For the management of the lesions resection of the pancreas (11 cases) or ligation of the bleeding vessel with external or internal drainage of the PC was performed (12 cases). Simple external drainage of a haemorrhaged PC in 3, and cystoduodenostomy or cystogastrostomy was performed in 5 cases respectively. Intraoperatively moderate bleeding gave some concern (7 cases), while post operatively pancreatic fistula developed in 9 patients drained externally. All stopped spontaneously. In two cases severe GI bleeding occurred post operatively. In both cases embolization of the bleeding vessels was performed successfully. No operative mortality occurred. The mean follow-up time was 40.6 months (5-106). Five patients died of unrelated causes, 3 patients underwent subsequent pancreatic operation, and 74.2% of the patients are doing well. Development of pancreatic PA was associated with a longer observation or conservative treatment period. Angiography should be considered whenever severe upper GI bleeding occurs in patients with known pancreatic disease and the source of bleeding is obscure. In selected cases selective embolization of the bleeding site may provide definitive treatment. PMID- 9408364 TI - Benefits of cell saver during the operation of gastric haemorrhage following repeated pancreatitis. AB - Authors report on a case of repeated pancreatitis causing several characteristic complications and concluded to splenic vein thrombosis, with an enlarged perisplenic vein network. The source of gastrointestinal bleeding is uncommon: originates from the submucous vein of the stomach. Cell saver was used under splenectomy because of great blood loss, and due to hyperimmunization occurred as consequence of massive transfusion. The benefit of cell saver was clearly evident from both point of view. PMID- 9408366 TI - The principle and practice of the minimal invasivity in the course of our traumatological work. AB - The authors examine which experiences can be deduced, in the course of their daily accident surgical work, from the minimal invasive chirurgical treatment of fracture contradicting many times the principles of the chirurgical treatment of fracture standardized by AO International. PMID- 9408365 TI - Evaluation of urinary enzymes in renal allograft damages by histology examination. AB - Acute allograft rejection (ARE) is one of the most current problem in kidney transplantation. Urinary enzymes (glutathione-S-transferase /GST/. dipeptidil dipeptidase /DPP/) are frequently used as prognostic factors of ARE. The authors compared the results of light microscopic study (by Banff scheme), and the GST, and DPP secretion in acute rejection. The correlation between the laboratory, and histology findings wasn't significant. our results suggest that both GST, and DPP are very sensitive, but less specific indicators in ARE, since their activity increases in many other damages as well. PMID- 9408367 TI - 10 years experience in performing Shouldice operation. AB - 206 male patients were operated for primary direct and indirect inguinal hernia, or both, by Shouldice technique at general surgical departments of Madadeni Hospital, Newcastle, South Africa, Paszto Hospital and Szolnok MAV Hospital, Hungary between 1986 and 1996. Mean age was 51 yrs +/- 15 yrs (17-91 yrs). The operations were performed by the original way of Shouldice described that type of hernia repair in 1945. 175 patients had spinal and 31 patients had local anesthesia with intravenous fluid and sedation respectively. Studies indicate that collagen metabolic dysfunction plays a major rule in the etiology of groin hernia. Until this is more clearly defined, surgeons will continue to repair groin hernias constitute 15% of operations in general surgery. In approach to groin hernia, the best view for examination of the inguinal region can be obtained by Shouldice technique to decide the proper surgical intervention to repair groin hernia. With low recurrence rate and rapid rehabilitation, author reports 2% of recurrence rate, the Shouldice operation highly recommended. PMID- 9408369 TI - Stereomatic constants of the biliary tree in normal functional state and during experimental cholestasis. AB - It has been found from the study of cholangiograms and laboratory specimens of biliary systems of human beings and dog's that the decrease in average diameters of biliary ducts during their passage from the hepatic portals to the periphery, is in accordance with the equation D = e-kz+b (z--sequence no of the duct, D- diameter of the duct, k--rate of change in the diameter, b--logarithm of diameter when z = 0). It has been established that parameter (b) defines size of the liver, and parameter (k) does not depend upon hepatic dimensions and changes during pathological processes. The use of (k) is recommended as an aid in defining the general character of dynamics of the duration of a given pathology. PMID- 9408368 TI - Dose depending effect of the complex action hemocorrector "Lactosorbal" on the motility of the distal colon in the paralitic ileus dogs. AB - For the model of paralytic ileus caused by the severe surgery it has been studied the influence of different doses of hemocorrector of "Lactosorbal" on distal colon motility. It has been established that stimulating effect of "Lactosorbal" on the motility of distal colon is of the dose-dependent nature. The results of the current survey are making the ground for multiple use of "Lactosorbal" to reach the most beneficial outcome of severe paralytic ileus resulting from major traumatic surgery in clinic. PMID- 9408370 TI - Is DNA aneuploidy a prognostic factor in gastric cancer? AB - Flow cytometric analysis of DNA ploidy was performed on an archival material obtained from 79 patients with gastric cancer who underwent a potentially curative (R0) stomach resection with D2 lymphadenectomy, and evaluated its relationship to conventional pathologic parameters, TNM stage and prognosis. No significant association between DNA aneuploidy and either patients' sex, depth of tumor infiltration (pT), lymph node involvement (pN), histological type according to Ming, macroscopic type according to Borrmann or tumor localization was found. However, the incidence of DNA aneuploidy was significantly lower in tumors of diffuse type according to Lauren, in tumors of signet ring cell or undifferentiated type according to WHO classification, in poorly differentiated/undifferentiated tumors and in patients younger than 50 years. We found no significant difference in survival between patients with DNA aneuploid tumors and those with DNA diploid tumors. Although the prognosis of patients with tumors of lower DNA index (DI < 1.2) tended to be better than for those of higher DNA index (DI > 1.2), the difference did not reach a statistically significant level (p = 0.09). TNM stage, depth of tumor infiltration (pT) and lymph node involvement (pN) were the only factors that significantly affected survival in univariate analysis and both pT and pN retained their independent prognostic significance in multivariate analysis. PMID- 9408371 TI - The use of modified Baylor score in the prediction of rebleeding in peptic ulcer hemorrhage. AB - The upper gastrointestinal bleeding is still an everyday problem. 40-50% of these bleedings are originating from peptic ulcer. The rate of rebleeding after initial hemostasis is 30-50%. In this group of patients we can observe the highest morbidity and mortality. The aim of this work is to select those patients who belong to the high risk group from the point of rebleeding. For this purpose we introduced after a retrospective analysis the modified Baylor score. In this scoring system the age, the number and severity of parallel illnesses, the hemostatus by the admittance, the ulcer size and location and the stigmata of recent hemorrhage are taken into consideration. On the basis of this every patient gets a score between 0 and 31. Based on our retrospective analysis we could establish three grades of risk groups: low risk (0-7), middle risk (8-11) and high risk (12 and over). In the low risk group there was no rebleeding. In the middle risk group we observed 4 rebleedings in 19 patients, while in the high risk group there were 32 rebleedings out of 36 cases. As a conclusion we can state, that the modified Baylor score is capable for the selection of high risk patients for rebleeding. With the early elective operations in these cases the high morbidity and mortality can be reduced. PMID- 9408372 TI - Investigation of microcirculatory changes in the duodenum of dogs caused by surgical suture materials. AB - Surgical suture materials play an important role in the safe performance of surgical interventions. In our experiments we made an attempt to investigate what microcirculatory changes result from pulling the thread through the wall of the duodenum in Lembert stitches by including 3 kinds of absorbable and non absorbable suture materials each (Catgut, Dexon, PDS, Silk, Ethibond, Ethilon). Research is still in the pre-experimental stage. In the long run, we hope to enrich the description of these suture materials by some new details. It could help prevent suture insufficiency, facilitate wound healing and thus, improved surgical safety. PMID- 9408373 TI - Laparoscopic deroofing of nonparasitic liver cysts. Comparison different methods. AB - Between 1982 and 1995 116 patients underwent operations because of NC of the liver, 18 of them were operated on by laparoscopic way. There were 94 male (mean age: 51.9 years) and 66 female (mean age: 54.6 years) patients. The mean size of the cysts were 57.4 mm in diameter (20-140 mm), In 94 cases they were solitary and in 11 they were multiple. In 31 cases we performed an enucleation, in 60 cases a fenestration and in one case a punction of the cysts. A liver resection had to be carried out in two cases. In 18 cases the fenestration of the cysts was performed by laparoscopy and in 10 cases cholecystectomy was simultaneously carried out. The most common complications were fever and wound suppuration. There was no mortality. Another 26 patients with NC have been treated with ultrasound guided puntion of the liver cysts. The cysts were solitary in twenty three cases and multiple in three other cases. The number of the cysts were thirty-two. One case had to be operated on because of recurrence of the cyst. Mortality could not be observed. In the laparotomized group the average time of the hospitalisation was 14.3 days, and among the patients who were operated on by laparoscopic way 7.0 days. An intervention is indicated only in case of severe complaints or the growing of the lesion. Recently we prefer the ultrasound guided punction or the fenestration of the liver cysts by means of the laparoscopic way. PMID- 9408374 TI - Host-versus-graft disease in mice after the induction of neonatal transplantation tolerance by two different methods. AB - Newborn A mice were injected either with a single i.v. dose (Group A) or with repeated doses of (B10 x A)F1 semiallogeneic spleen cells (SSC) (Group B). A similar degree of partial transplantation tolerance (TT) to B10 skin allografts was revealed in both groups. No signs of acute, rapidly fatal host-versus-graft disease (HVGD) (anemia, leukocytosis, severe thrombocytopenia, hepatic infarcts, gastrointestinal bleedings) were found, rather a chronic HVGD developed [moderate thrombocytopenia, autoimmune antithymocyte antibodies (ATA)] in both groups. The mortality due to lymphoproliferative disorders (LPD) was significantly higher in Group A. Thus, repeated perinatal injections of (B10 x A)F1 SSC into A mice did not increase the tolerogenic and the LPD-inducing effect either, and they did not elicit acute HVGD in contrast to observations in other F1 donor-recipient combinations [1]. Consequently, the development of acute HVGD depends on immunogenetic factors and not on the repeated administration of SSC. PMID- 9408375 TI - Surgical chemotherapy in the management of melanoblastoma of the lower extremities. AB - The therapy of advanced melanoblastomas of the lower extremities is limited. Surgery alone is insufficient due to the extent of the tumor, the radicality of mutilating surgery is questionable because of the existing or suspected subclinical metastasis. To avoid amputation, regional chemoperfusion and simultaneous hemofiltration may be the choice of treatment. Between 1993 and 1995 the authors performed surgical chemotherapy on 21 occasions in 14 patients with advanced melanoblastoma of the lower limb. Partial remission of 4 to 11 months developed in 10 patients, 3 patients achieved subjective improvement for 3 to 6 months, 1 patient had disease progression. Simultaneous application of surgical regional chemotherapy and hemofiltration offers an alternative approach in the management of patients suffering from advanced melanoblastoma. PMID- 9408376 TI - Tumor surgery of the pelvic region. AB - During the past ten years the authors operated on 27 tumors of the pelvic region, 12 of them involving the pelvic blade, 6 the periacetabular region, further 9 the os pubis and ischii, respectively. Most of the cases (16) were chondrosarcomas. The mean age of the patients--13 male and 14 female--was 41 years. As to surgical radicality 11 wide, 10 marginal and 6 intralesional resections were performed. After a mean follow-up period of 3 years (0.5-11 years) 19 patients are alive tumor-free, 2 with tumor, 4 died and 2 were lost to follow-up. As postsurgical complication wound-healing disorder and inguinal hernia occurred in 5 cases, on the surgical field thrombosis with secondary compartment syndrome and renal insufficiency developed in one case. The authors draw the attention to the difficulties and indications of the pelvic resections (internal hemipelvectomies). PMID- 9408377 TI - Cholecystectomy through a mini laparotomy alongside laparoscopic technique. AB - The authors present the surgical treatment of gallstones with special instruments and technique through a mini laparotomy, as developed in Kaposvar (Hungary). Despite the fact that the widespread use of laparoscopic cholecystectomy (LC) has succeeded over open cholecystectomy (OC), in their opinion, there are some instances where cholecystectomy through a mini laparotomy (MLC) would be advantageous. Last year (1996), in the authors' department, 32 MLCs were performed in addition to 72 OCs and 176 LCs. Due to unspecified techniques and differing criteria used to define MLC, it is difficult to find any data related to this subject in the literature available. In Hungary, there are two cities- Kaposvar and Szolnok--where only MLCs are performed for the surgical treatment of gallstones. Conversely, the University hospital of Szeged and the County hospital of Bekescsaba use all three procedures i.e. OC, LC and MLC; as is the case in the authors' hospital. In the article, the pros and cons of MLC are compared to those of LC. (The MLC being more cost effective, enables quick conversion to an OC if required, and provides the surgeon with a three dimensional picture, it is also suitable for the surgical treatment of common bile duct stones. However, it is cosmetically worse, obesity can cause technical difficulties, the incidence of hernia formation is higher, and wound healing is affected.) In the authors' opinion, it would be important to establish set criteria in order to screen patients that are unsuitable for LC, and they attempt to do so in this article. PMID- 9408378 TI - Continuous pO2 and pCO2 measurement in brain tissue and cerebrovenous blood during different inspired oxygen settings. A porcine model. AB - The clinical use of brain tissue oxygen measurement in patients with severe head injury is increasing. It is important to compare the findings in brain tissue with jugular bulb oximetry, to obtain normal values and to find out limitations of the method. We evaluated a newly available multisensor probe simultaneously in the brain tissue and in the sagittal sinus in a porcine animal model. We placed the Paratrend 7-probe (BSL, High Wycombe, UK) in the left frontoparietal white matter and measured pO2 (ptiO2), pCO2 (pti CO2), pH and temperature while simultaneously measuring these parameters (pcvO2, pcvCO2) in the sagittal sinus in 7 pigs under general anaesthesia during a 100% oxygen enhancement. The relation between oxygen increase in brain tissue and in the sagittal sinus showed a coefficient of correlation (CCmean) r(mean) = 0.96. The quantitative response in brain tissue was much more sensitive than in the sinus. A close correlation between pCO2 in brain tissue and sagittal sinus and the increase of the inspired oxygen was seen: CC ptiCO2 to arterial oxygen pressure (paO2) - r(mean) = 0.67, CC pcvCO2 to paO2 - r(mean) = 0.88. This is important for interpreting measured values and introducing new coefficients for patient monitoring. Newly available continuous brain oxygen measurement methods will allow better understanding of brain metabolism in the future. PMID- 9408379 TI - Micro and minilaparotomy surgery in the treatment of Mirizzi's syndrome. AB - The Mirizzi's syndrome presents a difficult surgical challenge because the dense adhesions and edematous inflammatory tissue cause distorsion of the normal anatomy in Calot's triangle, leading to a great risk of bile duct injury. Therefore, a controversial issue the surgical strategy for treatment of Mirizzi's syndrome since the introduction of laparoscopic cholecystectomy. The present study was undertaken to elucidate the applicability of microlaparotomy cholecystectomy in the management of Mirizzi's syndrome. PMID- 9408380 TI - The surgical technique of microlaparotomy cholecystectomy. AB - The authors are demonstrating the surgical technique of microlaparotomy cholecystectomy and the categorisation of minilaparotomy. In their 1575 unselected cases operated for cholelithiasis and their complications, 94% had microlaparotomy cholecystectomy (MLC: less than 4 cm single skin incision). 5.4% required modern minilaparotomy (4 to 6 cm incision) because of choledocholithotomy or bilioenteric fistula: and 0.3% had classical minilaparotomy (6 to 8 cm incision). In only 0.3% they converted into an incision longer than 8 cm. PMID- 9408381 TI - The removal of cystic duct and gallbladder remnant by microlaparotomy. AB - The so called "Postcholecystectomy Syndrome" may be due to various pathological biliary causes. While a very small number of patients may have symptoms attributable to problems related to cholecystectomy. Twenty five patients underwent a second operation on the bile ducts after cholecystectomy, cholecystostomy and choledocho-duodenostomy by micro and minilaparotomy between December 1990 and December 1996. The second most common causes for reexploration were cystic duct and gallbladder remnants (16%). After incomplete cholecystectomy they usually find that the cystic duct stump and the alot triangle embedded in inflamed scar tissue. For this reason the surgical risk is to high with laparoscopic surgery to reoperate for these pathological changes. PMID- 9408382 TI - The basic and the practical way of treating of diabetic foot. AB - Developed atrophic ulcer and infected alterations of the foot, as a result of complications of DM, according to the available literature, 40-80% of the performed amputations are not necessary even though in practice they do occur. In our practice even the severely altered and infected extremities which look serious are considered as primarily a savable extremity if the conditions are present. The state of circulation of the extremity and the condition of the limb are evaluated carefully, and the sugar level is monitored continuously. According to our experience, every progressing process, the alteration of the host sugar level can be held as responsible. That is why after the early therapeutic period, careful monitoring of the insulin level is a priority. At our out-patient department we are daily confronted during routine wound inspections by cases of necrotizing osteomyelitis which to our experience are doomed for removal. In 1995 at the out-patient section of the vascular surgery ward we saw over 9000 patients, more than 500 of which included patients with diabetic angio/neuropathy complications. The nursing of this group of patients during that year only to 33 large vessel reconstructive operations and 26 cases of amputations. We conclude from the above statistics that not all cases of osteomyelitis cases should lead to limb amputation. Due to the nature of the condition, careful monitoring with early preventive measures, plus family support play a crucial role in the outcome of the condition. This complex process is better handled if special diabetic centres were set-up to monitor patients progress. PMID- 9408383 TI - Oxygen free radicals and their clinical implications. AB - Reactive oxygen species (ROS) have been implicated in a variety of pathological processes. The generation of highly reactive oxygen metabolites is an integral feature of normal cellular metabolism (mitochondrial respiratory chain, phagocytosis, arachidonic acid metabolism, ovulation and fertilization), however their production can multiply during pathological circumstances. Free oxygen radicals act either on the extracellular matrix or directly upon cellular membranes themselves. The fundamental defense of the organism against ROS include scavenger enzymes (superoxide dismutase, catalase, glutathione peroxidase) and lipid- and water soluble antioxidant compound (ascorbic acid, glutathione, albumin, transferrin, etc.). Their role in ischemia-reperfusion models have now been comprehensively investigated and it has become clear that ROS is to be blamed for the bulk of post-ischemic injuries, hence the basis for newly established antioxidant therapy in such cases. Also more and more studies have concluded a pivotal role of ROS in degenerative and inflammatory conditions, post radiation processes and aging. Therefore it seems as we are continuously shedding light on the crucial part played by these molecules regarding a wide range of pathologies, we are discovering new therapeutic windows that would clinically assist us in managing such conditions. PMID- 9408385 TI - Surgery of the pancreas. AB - The author deal with the surgery of diseases of the pancreas in this article. On the basis of the literature, timing of operation, surgical procedures in acute pancreatitis are discussed. In chronic pancreatitis, results of different decompression operations, resection of the pancreas and endoscopic biliary stenting are evaluated. Regarding the carcinoma of the pancreas, operative methods and factors of survival are the most important questions. PMID- 9408384 TI - Cardioprotection during heart ischemia-reperfusion. AB - Oxygen reactive species play a significant role in reperfusion tissue damages. In this study we aimed to investigate the mechanisms of injury regarding changes of neutrophil function. In our experiments the left descending coronary artery (LAD) was ligated in Beagle dogs for 1 hour followed by one hour reperfusion. Animals were divided into two groups: Group I. dogs (n = 10) served as control: Group II. the animals (n = 10) were treated by cardioprotective drug Bisaramil. Peripheral blood samples were taken for neutrophil isolation before operation and subsequent reperfusion (5 min, 1 hour). The stimulated superoxide radical generating capacity of polymorphonuclear leukocytes (PMN) was measured. The lipid peroxidation (MDA), amount of reduced glutathione (GSH) and activity of superoxide dismutase (SOD) were measured in non-ischemic and ischemic parts of left ventricle. There was no significant changes either in control or in treated animals in respect to changes of neutrophil radical production after one hour LAD ligature, however there was a significant discrepancy (p < 0.001) between control and treated animals following a 1 hour reperfusion. The values of MDA in the ischemic-area increased characteristically in the Group I. parallel with decrease of scavenger GSH and SOD. In contrast in Group II., where depleted PMN radical production was observed endogenous scavengers were preserved on a higher level. In summary we can conclude that diminished superoxide radical production of circulating neutrophils during reperfusion has beneficial effects on tissue injury caused especially by free radicals. PMID- 9408386 TI - The role of high risk HPV lymph node positivity in the surgical staging of cancer of the uterine cervix. AB - The presence of oncogenic Human Papillomavirus (HPV) DNAs in cervical carcinomas and their normal and metastatic pelvic lymph nodes is studied worldwide. Does the HPV positivity without histological metastasis in the lymph nodes mean an early prognostic indicator in the progress of the disease? The authors analyzed the surgical, histological and virological data of four cervical cancer patients, whose recurrence occurred very early and died soon. After performing radical hysterectomy with precise removal of the pelvic lymph nodes, they assessed the operative specimens by routine histology and detected for HPV types by PCR. All the four lymph nodes harboured HPV-18 subtype in spite of negative histology in three cases. Thus the authors consider that if the lymph nodes contain any type of high-risk HPVs maybe despite negative histology or early stage of cancer we have to treat the patient as if she had an advanced cervical cancer. PMID- 9408387 TI - The importance of incision-planning and operation technique during organ preserving and radical operations in the case of patients with mammary cancer. AB - The authors place a great deal of emphasis on the application of plastic surgery principles in the course of all their operations, especially mammary operations: the incision planning respecting the lines of strength, the atraumatic operation technique (sharp preparation, high-frequency electrocoagulation, tensionless wound closure, and suction drainage from a separate opening). In their organ preserving operations the radial incision recommended by Veronese is avoided, and the axilla is exposed separately. They do not perform their radical mammary operations according to the customary method (Auchincloss, Patey) either. If necessary they accomplish the tensionless closure of the wound by means of individual incision planning and the preservation (rotation and sliding) of the intact skin of the breast, which is the precondition of the reconstruction that might be performed subsequently. In the course of the follow-up of 295 operations carried out in this manner excellent or good results were obtained in 83% as far as aesthetics and functionality were concerned. Lymphedema did not develop in a single case on account of operation technique. PMID- 9408388 TI - The totally extraperitoneal (TEP) laparoscopic hernia repair. AB - In recent years endoscopic techniques using mesh implantation have been added to the many options for the repair of inguinal hernia to diminish postoperative pain, shorten the reconvalescence period and improve the recurrence figures of the classical repair. The purpose of this paper is to evaluate our first experiences gained by applying the TEP laparoscopic hernia repair. Between March and December 1996, 20 laparoscopic herniorrhaphies were performed with complete extraperitoneal balloon dissection. A large polypropylene prosthesis was inserted to cover all potential defects. The follow-up was 2-10 months. There were 10 indirect, 6 direct, 1 combined direct and indirect, 1 femoral and 2 scrotal hernias. Age (26-86 years) and operative time (52-120 mins) had a wide range. Hospital stay lasted from 1-5 days. Morbidity was low: scrotal emphysema (3), peritoneal lesion (2) and palpable mesh crease (1) occurred in a few cases. No recurrences have been seen so far. It seems that the TEP laparoscopic hernia repair is a highly successful procedure with minimal morbidity. Preliminary results are promising. Further experiences and long term follow-up studies will determine the future of laparoscopic hernia surgery. PMID- 9408389 TI - The trental treatment prevents the reduction of superoxide dismutase enzyme (SOD) activity in rats after colon surgery. AB - The authors tested the effects of the pentoxiphyllin (px, Trental) pre-treatment upon the superoxide dismutase activity of the left colonpart anastomosis of rats. It has been found that the SOD activity of the proximal (4.9 U/g), the anastomical area (1.9 U/g) and the distal (3.1 U/g) intestinal segments considerably decreased, compared to the control (9.16 U/g). In the operated and pre-treated animals with Trental, the SOD activity of all the three intestinal segments increased (proximal 15.5-anastomosis 5.7-distal 8.9 U/g) compared to the non treated group. PMID- 9408390 TI - Changes of volume substitution for the treatment of hemorrhage within 10 years and their costs. AB - Whereas 10 years ago blood loss was substituted by whole blood application together with colloidal or crystalloid substitutes, this behavior changed to a distinct therapy according to the changed laboratory parameters with the application of red cell concentrates together with fresh frozen plasma (FFP). By means of two representative operations (resection of sigma and hemicolectomy) the behavior of volume application and the substitution of blood and blood components were controlled for the years 1980 and compared with the behavior of 1990. In addition to that the costs for both behaviors were calculated and compared. The volume substitution of altogether 176 patients was investigated. 87 patients were operated in the year 1980 and 89 patients in the year 1990. 60% sigma resection were observed on both decades. The analysis revealed that the main substitute (60%) was whole blood in the year 1980 whereas in the year 1990 only 3% of the patients were treated with whole blood. A reverse development was observed with the application of red cell concentrates which was only 2% in the year 1980 but 54% in the year 1990. Unexpectedly the consumption of FFP remained nearly constant in both decades whereas the administration of 5% albumin increased from 40% to 80%. Also the behavior with regard to colloidal substitutes changed markedly within the 2 decades. Dextran and gelatin preparations were exclusively applied in the year 1980 and starch preparations in mainly the year 1990. This changed behavior was responsible for an increase of the costs of 100 $US for every patient. Although the changed behavior can be explained with advantages for the patient but this must be paid by an increase of the costs. PMID- 9408392 TI - In ophthalmology new possibilities for the use Nd:YAG laser. AB - Results of the technical development in laser technology created new possibilities in ophthalmic surgery, too. The easy access to various lasers offers an extraordinary opportunity for eye surgeons. At the Department of Ophthalmology of the University Medical School of Debrecen we performed 1400 laser treatments in 1996. Out of that 1100 were retinal Argon laser coagulations because of diabetic retinopathy, 200 YAG laser capsulotomies because of secondary cataract, and 30 laser iridotomies because of glaucoma. Additional 70 laser treatments were performed for special indications. During the laser treatment of the retina and the choroid we use Argon gas lasers, whereas the Nd:YAG laser used in the anterior segment of the eye is a solid-state laser. Our pulse mode modus synchronized Nd:YAG apparatus can produce megawatts of energy in nanoseconds. Authors give an account of the Nd:YAG lasers' possibilities in therapy (capsulotomy, iridotomy), present the most frequent indications of treatments in 1996, and compare it with international literature data. Authors demonstrate the newest principles of the Nd:YAG laser treatment of secondary membranes that are occasionally formed in the anterior chamber especially in the area of the pupil following cataract surgery. PMID- 9408391 TI - Free radical reactions in the gallbladder. AB - The changes in the composition of bile can lead to the process of it's crystallization in the gallbladder. In bile model it was shown that inflammation with the generation of reactive oxygen metabolites may induce and influence the cholesterol monohydrate crystal formation within supersaturated bile. The aim of this study was to investigate the ability to detect traces of reactive oxygen metabolites, thiobarbituric acid reactive compounds and dien, in order to compare cholesterol and bilirubin contents in bile and serum during different conditions of inflammation in the gallbladder's wall. In every bile sample a reference to free radical reaction was found. There was an increase in MDA during higher degree of inflammation in the gallbladder, but no alteration in the dien content was observed. In case of common bile duct stones the bilirubin in the serum and in the gallbladder was parallelly high, but in other cases there were no significant correlation. In an occluded gallbladder with hydrops the content of protein was significantly higher in 85% of the cholesterol stones. As a conclusion, free radical reactions in the wall of gallbladder as well as in bile can induce gallstone formation. Further studies are needed to clarify the time which is sufficient to change the composition of bile and the degree of inflammation which lead to the onset of stone formation. PMID- 9408393 TI - Mucosal permeability changes during intestinal reperfusion injury. The role of mast cells. AB - The objective of this study was to investigate the role of intestinal mast cells in mucosal functional and morphological alterations induced by 30 min segmental ischemia and 120 min reperfusion in anesthetized dogs. The time course of permeability changes of the mucosa to sodium fluorescein (NaFl) in blood-lumen and lumen-blood directions was studied in two separate series of experiments. Local hemodynamics, intramucosal pH (pHi) alterations, mast cell number and degranulation and the degree of tissue injury were determined. The effects of cromolyn (peritoneal-type mast cell stabilizer), quercetin (mucosal-type mast cell stabilizer), and dexamethasone (aspecific membrane stabilizer and mast cell depleter) pretreatments were evaluated. Ischemia-reperfusion induced significant tissue injury, elevated segmental vascular resistance, and decreased pHi. The blood to lumen clearance of NaFl increased significantly during ischemia and reperfusion. Cromolyn and quercetin pretreatment significantly inhibited permeability changes, but did not influence pHi and morphological alterations induced by ischemia-reperfusion. Dexamethasone pretreatment did not influence the number of mast cells, however, the degree of mast cell degranulation and the degree of mucosal damage decreased. These results demonstrate that mast cells or mast cell-induced reactions contribute to the mucosal permeability alterations and barrier lesions during reperfusion, but play a minor role in reperfusion induced structural injury. PMID- 9408394 TI - Rare case of the utero-vesical fistula caused by intrauterine contraceptive device. AB - The vesico-uterine fistula is a very rare disease. There have only been 150-200 causes. We are reporting on a case in which the chronic fistula was caused by an IUD having been placed 4 years ago and it "wandered" through the bladder. A 30 year-old patient in 1992 and IUD was inserted. She had gynecological controls twice, in 1993 last time. She has problem of urination very often. A cyclical bladder bleeding drew the attention to the disease. In ambulanterely performed cystoscopy we found an IUD perforating towards the interior of bladder in the borderline of its bottom and back wall and was situated in the bladder with its 3/4. We have removed it with forceps. After six weeks of expectation and strict observation did we want to manage the fistula after having consolidated the symptoms of the inflamed surroundings. During the operation we have noticed a wallment size mass of scar between the uterus and the bladder expanding to the height of the orifice of the uterus. The scarily fixed bladder has been separated from the cervix and the scarry wall of the fistula has been cut out. We have brained the cervix towards the vagina and then we've sutured the cervix and the bladder with Dexon 'O' treat, as well. We have interposed a surgical net between the cervix and the bladder followed by blood-clotting and peritonisation. We should take the follows into consideration: careful separation, fine operative technique, and strong well absorbing thread as well as trying to keep the organ. In our opinion the bioplast--interpositum used on our case maker the efficiency of the operation higher. PMID- 9408395 TI - Endothelin-1-induced circulatory response in the rat: the role of ETA and ETB receptors. AB - Production of the powerful vasoconstrictor endothelin-1 (ET1) is increased in a number of pathological conditions. This study was performed 1. to assess the effects of a twofold elevation of circulating ET1 on global hemodynamics and cardiac function, and 2. to determine the ET receptor subtypes that are responsible for this action. We have used the ETA receptor-selective antagonist BQ 610, the novel ETA receptor antagonist ETR-Pl/fl peptide and the specific ETB receptor antagonist IRL 1038 to investigate the role of these receptor subtypes in mediating circulatory changes induced by ET1 in anesthetized Wistar rats. ET1 infusion produced a significant rise in mean arterial pressure (MAP), elevated total peripheral resistance (TPR), and decreased cardiac output (CO). BQ 610 and ETR-Pl/fl pretreatment significantly attenuated the ET1-induced hemodynamic changes. Pretreatment with IRL 1038 had no effect on CO, but significantly reduced MAP and TPR elevation 20 min after ET1 infusion. These results suggest that ET1 may contribute to circulatory failure in conditions with increased ET1 production via a mechanism involving ETA receptors. ETB receptors, albeit to a lesser extent than ETA receptors, are also involved in mediating ET1-induced peripheral vasoconstriction in the rat. PMID- 9408396 TI - Idiopathic scoliosis--new surgical methods or search for the reasons. AB - Idiopathic scoliosis (IS) is a deformity of the spine whose aetiology is unknown despite of extensive clinical and basic research work. The significance of this disease lies in the consequences of a spine deformity (deformation of the chest, compromise of the cardio-pulmonar function, compression of the neurological elements, severe somatic and psychological sufferings). According to statistical data, one third of the idiopathic scoliotic cases progress and need surgical intervention. Cotrel and Dubousset introduced the three dimensional correction surgical method for the treatment of spine deformities in 1983 (CD method). The authors report on the application of the CD method for the treatment of IS from 1991 to date. After the operation of 36 patients they give an account of good results and report on the modifications of the surgical technique on the basis of their experiences. Aware of the late consequences of the surgical treatment of IS the authors started with the investigation of the aetiology of the disease in order to introduce a causal treatment. As for the aetiological background of IS, data on the role of heritable factors were relevant only. They gave a hint to start with studies aimed at the investigation of genetic determinants of the syndrome. I aberration of chromosome #10 [karyotype: inv(10)(p11q26)]. In one of the cases there was a multiple familial occurrence of the syndrome coupled with the same anomaly of the karyotype. The probands' father and grandfather all had the same pattern of scoliotic deformity and the same aberrant karyotype of chromosome #10 was found both in the proband and his father. Preparation of genomial DNA from peripheral blood of the patients was started. Fragment length polymorphism (RFLP) examination with PCR technique using oligonucleotide primers specific for chromosome #10 and for regulatory genes of left-right body assymetry is underway. PMID- 9408397 TI - Predictive morphological findings in "zero-hour" biopsies of renal allografts. AB - "Zero-hour" biopsies of 65 donors have been performed since 1994. Donor kidneys were categorized into five groups based on the morphological findings in "zero hour" biopsies. No morphological abnormalities were found in 38% of the cases (group 1). Arteriosclerosis was present in 31% of donor kidneys (group 2). Specific morphological alterations, i.e. acute tubular necrosis [21.5%], tubulointerstitial nephritis [6.2%] or glomerulonephritis [3.1%] were detectable in the cases remained (group 3-5). During an average of 336 posttransplant days clinical and histological follow up was performed (50 rebiopsies). Statistical data of mismatch (1.4-2.0), average of donor/recipient age (35-42 years), cold and warm ischaemic time (1290 and 66 min) were comparable in all groups. According to our observations: 1. higher creatinin was found in grafts with arteriosclerosis (group 2) (p < 0.05), 2. there were more non-viable grafts and longer period of delayed graft function in acute tubular necrosis (group 3), 3 higher creatinin, rejections with the need of rehemodialysis were observed in four cases of tubulointerstitial nephritis (TIN-group 4). Glomerulonephritis (GN group 5) grafts had only delayed graft function, however these groups were few for statistical evaluation. Biopsy complication in 1/115 cases was found (rebiopsy induced kidney haemorrhage). In conclusion, "zero-hour" biopsies can be useful and safe tools to predict early graft function. Besides "zero-hour" biopsies help the histological interpretation of consecutive graft rebiopsies. PMID- 9408398 TI - Splenic autotransplantation after abdominal trauma in childhood. Clinical and experimental data. AB - Splenectomy is known to increase the risk of overwhelming bacterial infection. There is a decrease in immunoglobulin IgM and T-lymphocytes, primary antibody response to antigen challenge is impaired, altered opsonic function an Tuftsin deficiency are noted. Splenic autotransplantation has been suggested as a method of preserving function and this concept is supported by experiments in animals. Prior to operation on humans the technique was thoroughly elaborated and practised in animal experiments (dogs). After splenectomy, 6-8 thin segments (Furka's "spleen chip") are placed in between the plates of the major omentum. Within the period of ten years out of 52 patients 11 children (4 girls, 7 boys) suffered from abdominal trauma underwent total splenectomy, and than autotransplantation in the Kenezy Hospital in Debrecen, Hungary. In several patients the postoperative follow-up radionuclide imaging, IgM, and Tuftsin levels, and the haematological changes (leukocytes, differential blood count, platelet count, iron level in serum) unambiguously confirmed the function of the splenic tissue. PMID- 9408400 TI - Efficacy of prevention of thromboembolic complications with LMW-heparin in experiment. AB - The authors used an antithrombotic agent (Nadroparin Calcium) with anti-Xa effect in their experiments to prevent thromboembolic complications in the model of endoprosthetic replacement of the hip joint in mongrel dogs. 10 experimental animals (Group I.) were given doses of 100 A Xa ICU/kg/bwt of Nadroparin Calcium subcutaneously 4 hours prior to the operation and also once a day until the 3rd postoperative day; between the 4th and 10th postoperative days doses of 150 A Xa ICU/kg/bwt Nadroparin Calcium were given. The 10 control animals (Group II.) did not receive anticoagulant treatment. In both groups platelet count, activated partial thromboplastin times (APTT), prothrombin and fibrinogen levels as well as activated factor X inhibition (F Xa) were measured prior to surgery and also until the 14th postoperative day. No changes in APTT and prothrombin levels were detected during the experiment, however platelet count and fibrinogen levels as well as the extent of F Xa inhibition showed significant and different changes in groups I. and II. The Group I. which had received thromboembolic prophylaxis did not develop deep venous thrombosis or pulmonary embolism, but the control group did. Based on their investigations, the authors concluded that they had been able to achieve F Xa inhibition by giving the above mentioned doses of Nadroparin Calcium which was enough to prevent thromboembolic complications in their model experiment of implanting hip endoprosthesis. PMID- 9408399 TI - Mini-sternotomy for aortic valve surgery. AB - In the recent years more and more efforts have been made widely to introduce new techniques in the minimally invasive cardiac surgery. At our Department and the Linkoping University Hospital, Cardiothoracic Surgery Department, from August 1996 to January 1997, aortic valve surgery was performed in 23 adult patients (9 female, 14 male), age 28-86 years (mean age 62.5 years). Twenty-two patients had aortic valve replacement, among these, in 3 cases concomitant aortic annulus dilatation was made and in one case reduction-plasty of the dilated ascending aorta. In another one case resection of a sub-aortic membrane was performed. The operations and postoperative period were free of complications in all patients. Following an average 36 hours intensive care all patients were discharged after an average of 11.2 day hospital stay. The authors introduce the new surgical technique and present its advantages and disadvantages. Mini-sternotomy has less detrimental structural and functional effects on the thorax. Moreover, due to its minimal surgical trauma, this less invasive technique reduces patient morbidity, hospital stay and cost of care. Since mini-sternotomy is a safe and advantageous technique, the authors recommend applying this new technique in most of aortic valve operations. PMID- 9408401 TI - Laparoscopic distal resection of the pancreas with the preservation of the spleen. AB - Management of the pancreatic diseases is still a challenge to the laparoscopic technique. Some experience has been gained in the laparoscopic exploration of the pancreas and staging in cancer. Anatomically the accessibility of the distal pancreas provides the laparoscopic approach technically feasible. PATIENT AND METHOD: A case of insuloma in the tail of the pancreas is presented, where distal pancreatic resection was performed laparoscopically with the preservation of the spleen. In a 55 years old female patient with typical clinical symptoms of hyperinsulinism CT identified a 3 cm large solid tumor in the tail of the pancreas. Complete mobilization of the distal pancreas was enhanced by the use of an ultrasonic dissector (UltraCision). The pancreas is detached from the splenic hilum after dividing the spleen vessels. The pancreas is transected proximally by laparoscopic linear stapler. Preservation of the short gastric vessels provides the necessary blood supply of the spleen following division of the splenic artery and vein. Thus removal of the spleen is not a necessary step in this procedure. The operation was carried out within 4.5 hours. Postoperative course was uneventful, the patient left the hospital on the 5th postoperative day. Advantages of the procedure were the earlier mobilization and shorter recovery time, less postoperative pain. The procedure can be safely performed with a good experience in both pancreatic and laparoscopic surgery. PMID- 9408402 TI - Intracellular free Ca2+ elevations in cultured astroglia induced by neuroligands playing a role in cerebral ischemia. AB - For better understanding of glial participation in cerebral ischemia, spectrofluorimetric analysis using the calcium indicator Fura-2AM was applied to examine the role of intracellular free Ca2+ ([Ca2+])i elevation induced by different neuroactive substances in cultured rat brain astrocytes. The activation by the general receptor agonist glutamate resulted in a biphasic cell response in [Ca2+]i. We couldn't observe N-methyl-D-aspartate-evoked [Ca2+]i response at all. Quisqualate triggered a complex [Ca2+]i response in astrocytes consisting of mobilization of Ca2+ from the intracellular stores and also Ca2+ influx from the extracellular space. Kainate elicited a markedly different Ca2+ signal an external Ca(2+)-dependent sustained [Ca2+]i rise resulting from the activation of the ionotropic glutamate receptor. According to our results two types of glutamate receptors, the quisqualate-specific metabotropic and kainate-specific ionotropic receptor, are involved in [Ca2+]i elevation in these cultures. We could monitor agonist-specific cell response to noradrenaline, serotonin, vasopressin and ATP as well in these cultured rat astrocytes. PMID- 9408403 TI - Splenectomy and spleen autotransplantation due to splenic cyst. AB - Splenectomy is known to increase the risk of Overwhelming Postsplenectomy Infection (OPSI). Autotransplantation is one of the possibilities to preserve splenic functions. The authors performed spleen autotransplantations in two cases after the splenectomy of cystigerous spleens. Ten pieces from the intact spleens each sized 10 x 20 x 2 mm were implanted in between the plates of the great omentum according to Furka's spleen apron method. The function of the reimplanted splenic tissue was demonstrated by scintigraphic investigations. PMID- 9408404 TI - Monitoring of gastric acidity following ductal decompression surgery for chronic pancreatitis. AB - The accepted decompression methods of chronic pancreatitis are the longitudinal pancreaticogastrostomy and the conventional pancreaticojejunostomy. The aim of the present study was to estimate the effect of these types of drainage operations on gastric acidity and to evaluate the clinical results. Between Jan. 1992 to 1996 56 patients with chronic pancreatitis were selected into the investigation who were operated in our clinic. A 24 hour gastric monitoring was taken on every patient before and 6 weeks after the operation. Following a complete postoperative check up we found that both types of operations are effective for pain relief (71%). Retrospectively 83% of the patients had no digestive problems due to pancreatic enzyme substitution. According to our statistical evaluation of 24 hour gastric pH monitoring test no alteration was detected in gastric pH in both groups pre- and postoperatively. On the basis of pH measuring and evaluated data we consider that pancreaticogastrostomy is a good operation choice to relieve intractable pain in selected patients with chronic pancreatitis associated with duct dilatation. PMID- 9408405 TI - Inflammatory mediators and surgical trauma regarding laparoscopic access: neutrophil function. AB - Polymorphonuclear leukocytes (PMN) are well recognised as being the principal cells in the inflammatory response reaction. The current investigation was designed to evaluate the stimulated state of neutrophils in patients undergoing open (OC) and laparoscopic cholecystectomy (LC). The superoxide radical generation of isolated PMN and its lag time, and PMN elastase were measured in peripheral venous blood samples collected from 42 patients. The observation period started on the day of hospital admission and ended on the 5 th day. Our results showed that although there were no characteristic changes in the superoxide radical production of PMN in the LC group, we found a marked depletion already by the first day in OC group which persisted during the whole observation period. There was an elevation of PMN elastase on the first postoperative day in both groups, but in the laparoscopically operated patients it decreased considerably on the 3rd postoperative day while in the OC group it maintained a high level. The same discrepancy was present between the two groups on the 5th day. Decreased free radical production of isolated PMN may reflect depleted neutrophil functional state, while changes of PMN elastase indicates increased harmful potential. These data suggest the differences in neutrophil activation between the two groups can be partially responsible for the better outcome in patients operated by laparoscopic cholecystectomy. PMID- 9408406 TI - Postoperative chemoembolism treatment of primary and metastatic malignant hepatic tumours. AB - Between 1992 and 1996 62 patients were operated for primary malignant and metastatic hepatic tumours. After the operation 31 patients received cytostatical treatment, 9 patients were treated with the method of chemoembolism. 46 patients are alive 30.7 (5-54) months after the operation. 9 patients died 12.5 (3-27) months after the resection of the liver. Authors have favourable experiences on the treatment of chemoembolism. PMID- 9408407 TI - Surgical treatment of diverticulitis of the sigmoid. AB - This study demonstrates that elective colon surgery for diverticular disease is benign, with a low mortality and morbidity. The acute severe complications of the colonic diverticular disease are characterized by a large mortality and morbidity. The results reported here, as all those published recently on the same subject, are in favor of broadening the indications of colectomy in diverticular disease, before the occurrence of a severe acute complication. PMID- 9408408 TI - Difficulties in early diagnosis and treatment of uncommon breast tumours. AB - The aim of the current study was to analyse the uncommon histological type tumours, occurred in our department in the last 10 years. In the study period 921 patients were treated for breast malignoma, 47.4% of them in early cancer stage. In the latter group 3 cases were observed with rare histological tumour type: secretory juvenile breast cancer T1N0M0, GI /case I/; primary angiosarcoma T1N0M0, GI /caseII/; planocellular breast cancer T2N0M0, GII /caseIII/. In the case of rare histological type tumours: (1) triplet diagnostic procedures are usually able to result in correct preoperative diagnosis; (2) the adequate treatment has to be determined individually; (3) continuous, long term follow-up is necessary; (4) MRI is an appropriate technique in the detection of recurrences. PMID- 9408409 TI - Different opening techniques in cataract surgery. AB - A retrospective study on the different opening techniques in cataract surgery at our Department in 1996 was carried out. In our everyday practice fornix based conjunctival flap and sclerocorneal oblique or vertical-oblique approach with a center at 12 o'clock was performed. In the case of hemorrhagic diathesis and secondary ACL implantation the corneal approach was preferred. After glaucomatous filtration surgery the incision was made temporally in some cases. PMID- 9408411 TI - Two methods in the treatment of prostate cancer T1-T2. AB - The author reports experience of 82 radical retropubic prostatectomies. In accordance with the length of isolable urethra two methods were applied. In one of the methods the bladder-neck was pulled down to the floor of the pelvis by a synthetic reabsorbable thread (40 cases), in the other method, the urethra-trunk was sewn up to the bladder-neck (42 cases). The early and later complications are dealt with. Their findings: the blood transfusion during intra and perioperative period was identical in both methods. Injury of ureter did not occur, injury of rectum in one case (1.2%) occurred. In the two methods incidence of impotence (85%) and incontinence (15%) were identical. The length of operation was on an average twenty minutes shorter in the pull-down technique but stricture occurred in 4.7%. In the sewing method no stricture occurred. In the opinion of the author the pull-down technique at the short (5-7 mm) urethra-trunk, while the direct sewing-up technique at the long (7-20 mm) urethra-trunk were applied. PMID- 9408410 TI - New indications for and technics in vitreoretinal surgery. AB - Vitreoretinal surgery has become a widely used ophthalmic surgical procedure since the late sixties. However, it remained an intensively developing field of surgical interventions, and indications still become wider and wider. The procedure starts with the removal of vitreous body, blood or inflammatory material from the vitreous cavity. After this, the given retinal disease, such as retinal detachment, macular hole, proliferative diabetic retinopathy, intraocular foreign body, etc. can be properly treated in situ. For that purpose a number of fine intraocular instruments, fluids, gases can be used. While the anatomical success rate can be up to 95%, the final visual outcome, due to the fact that the retina is not rarely seriously damaged, is less enthusiastic. However, a great number of previously untreatable vitreoretinal diseases can be managed in this way, nowadays. PMID- 9408412 TI - Experiences with ethanol infiltration of the liver metastases of colorectal tumors (preliminary study). AB - With the aid of improvement in imaging technique and with the ability of detection of hepatic tumours, the successful treatment of primary and secondary hepatic tumours using surgical methods has gained importance. Metastasectomies were performed in 16 cases at our department in 1996. In 6 cases atypical resections and/or infiltration with 96% ethanol were performed. In 3 instances ethanol infiltration into the metastases were accomplished during the resection, in 4 cases--when metastases were discovered 1-4 year after the resection of the primary tumours--the same method was used during the relaparatomy. 96% ethanol was injected directly into the metastatic tumours during operations. The early ultrasonograph examinations proved the total destruction of the metastases in diameter of 1-3 cm, and partial necrosis of the large (d = 3-8 cm) tumours. Two patients were lost from the follow up. Five patients are still alive after 2-8 months of the ethanol infiltration. In one patient after the ethanol infiltration of metastases /d = 3-5 cm/ in 6 segments of the liver we observed the dissemination of the tumour. PMID- 9408413 TI - The cryovaricectomy (demonstration of a new method). AB - The operating technique of the primary varicosity has changed recently. The subcutaneous methods have been used nowdays. We give account of making 416 cases by cryovaricectomy, which is a quite new method. Using this technic we can remove the dilated venous branches during a 3-4 millimeters wide incision of skin and preserve the sufficient main vein. Summarizing of our advantageous experience: the cryovaricectomy is suitable for principles of minimally invasive surgery, and the operating time is much more briefer, the aesthetic results are best, the attendance far shorter and the postoperative complaints of patients much more fewer. PMID- 9408414 TI - A randomized comparison between minilaparotomy and conventional approach for aortoiliac reconstructive surgery. AB - To decrease the surgical stress on patients undergoing aortic surgery the authors developed a less extensive procedure utilizing minilaparotomy and videoendoscopy. From June 1993 through July 1996, patients undergoing surgery for aortoiliac occlusive disease were randomized comparing the minilaparotomy (ML) with conventional approach (CA). Sixty-two patients participated in this trial, with 37 in the ML group and 25 in CA group. There were no significant differences between the groups in terms of age, sex or comorbid conditions. The incidence of intraoperative complications was similar for both groups. After surgery, nasogastric drainage was significantly (p = 0.01) shorter, bowel movement and initiation of alimentation began earlier in ML group. Both groups of patients showed a significant decrease in vital capacity and forced exspiration volume postoperatively, but this depression was significantly higher (p = 0.05) in CA group. The ML group also had significantly shorter stay in the intensive care unit (p = 0.001) and the mean duration of the postoperative hospital stay was also significantly less (p = 0.05). Generally, the patients operated by ML method during the postoperative recovery period required less analgesia, and expressed better "overall satisfaction". In long-term follow-up (mean 21 months), there was no significant difference in survival rates between two groups. PMID- 9408415 TI - Effects of nitric oxide synthase inhibition on the hemodynamic changes in hyperdynamic endotoxemia. AB - In this study we compared the circulatory effects of the arginine analogue non specific nitric oxide synthase (NOS) inhibitor N omega-nitro-L-arginine (NNA), and the specific inducible NOS (iNOS) inhibitor S-methylisothiourea (SMT) and S (2-aminoethyl)-isothiourea (AEST) in a hyperdynamic endotoxemic dog model. Mean arterial pressure (MAP), cardiac output (CO), and myocardial contractility (MC) were measured. A hyperdynamic circulatory response was elicited with a 2-h infusion of a total dose of 5.3 micrograms/kg E. coli endotoxin (ETX). NOS inhibitory treatment (2 mg/kg) was administrated from the 45th min of endotoxemia. ETX induced a hyperdynamic circulatory response, and a significant myocardial depression. NNA induced a prolonged, SMT a transient increase in MC, both drugs elevated MAP, but decreased CO. AEST significantly prolonged the elevation in CO, but did not affect MAP. Selective inhibition of the iNOS may be a beneficial in sepsis. PMID- 9408416 TI - Reconstruction of the constricted orbital cavity in patients with artificial anophthalmy. AB - In cases, in which not only the globe of the eye, but the whole conjunctival sack is missing and the eyelids are due to scarring attached either to the orbital tissues and/or to each other, it is not possible to insert a prosthesis. This is a major aesthetic defect. The cavity that allows artificial eye insertion can be formed by surgery. One of the best methods is the orbital cavity reconstruction developed and suggested by the Hungarian Istvan Csapody. He constructed two instruments to perform the operation: a butterfly shaped marker to prepare skin graft with proper size and shape, and an adjustable cone formed spacer to keep the reconstructed socket open during the healing process. We use free autologous split skin graft to line the cavity. If the shapes of the eyelids are intact, and the color of the prosthesis matches the fellow eye and it fits perfectly, then the postoperative cosmetic result can be excellent. PMID- 9408417 TI - Treatment of ingrowing toenail with segmental chemical ablation. AB - During a 5 year period (1992-1996) 716 segmental chemical ablations using phenol solution of 80 v/v % were carried out because of ingrowing toenail. As a control, Emmert procedure was done in 200 cases. In the present study we discuss the etiology, the surgical technique of the chemical ablation and the results of the intervention. We have found that phenol treatment results in a larger rate of symptomatic relief, lower recurrence rate and better cosmetic results could be achieved. Taking into account the good results, the low financial requirements of the procedure in the treatment of ingrowing toenail, segmental chemical ablation with phenol may be the therapy of choice of the disease. PMID- 9408418 TI - Shall we wait until the head of the femur is completely crushed in idiopathic necrosis of the femoral head? AB - Although it is of polyetiological nature but of identical pathomechanism, the necrosis of the femoral head has been known for more than a century, and, without treatment, it results in complete necrosis. To stop the progression of the process, load-lifting drilling of the affected area was introduced in the 1970s. Since then, the success of the technique in the early stage of the disease has been justified by countless follow-up examinations and studies. Load-lifting drilling of the affected area in the early stage of the disease yielded similar results at our department in a small clinical sample over 6 years. To ensure permanent drainage, we have used cannulated titanium screws with a diameter of 9 mm for the second year now; besides allowing continuous decompression these screws hold mechanically well and the use of titanium allows MR and CT investigations later on. PMID- 9408419 TI - About the primary malignant tumors of small bowel. AB - Primary malignant tumors of small bowels are relatively rare among the GIA tumors. There are no specific methods to find these tumors in an early stage. Only the specific radiological examination (double contrast method or CT-scan) or sometimes the specific jejunoscopy can prove the correct diagnosis before the operation. Usually surgeons have to operate because of the emergency situation caused by primary malignant tumors of small bowel. A total of 21 patients were operated on for malignant tumors of small bowels since 1st January, 1986 till 31st December, 1995 (during 10 years). 13 of the 21 were urgent cases. 3 perforations and 10 mechanical bowel obstructions were the indications of the acute surgical procedures. The preoperative diagnoses were correct in the eight elective operations. Types of operations: 8 jejunal-resection, 11 ileal resection, 1 Whipple-operation, 1 ileal-resection with right hemicolectomy. Peri operative mortality rate was zero. Histological reports: 13 adenocancers, 2 leiomyosarcomas, 5 malignant carcinoids, 1 anaplastic cancer. Lymphomas and tumors of papilla Vateri were excluded from this study. Most of the patients had abdominal symptoms earlier than 8 months before the operations. Earlier diagnosis may give a better chance to operate these tumors in earlier stages, and it can give a better chance for the patients for longer survival. PMID- 9408420 TI - Locked gates: profit and pain. PMID- 9408421 TI - Emergency medicine research: where are we now and where do we need to be? PMID- 9408422 TI - Ambulance transports: what are the alternatives? PMID- 9408423 TI - Computed tomography scanning and delirium in elder patients. AB - OBJECTIVE: 1) To examine the ordering of head CT scans in elder patients with delirium and cognitive impairment; and 2) to report CT scan findings associated with these conditions. METHODS: This was a 2-part study. Part 1 was a prospective, observational study of 560 adults > 70 years of age evaluated at 3 separate EDs using a 200-hour stratified sampling process at each ED. During Part 1, the frequencies of specific findings (i.e., delirium, impaired consciousness, and impaired cognition) and CT scan rates for these groups were determined. Part 2 was a retrospective analysis of CT scan reports and medical records (n = 279) for patients > 70 years of age in the prospective sample (n = 79) and from a sample (n = 200) of CT scans obtained at a fourth ED. Part 2 examined clinical findings detected in the ED to determine those factors that were associated with acute findings on CT scan. RESULTS: Part 1: There were 333 (59.4%) patients prospectively classified as having impaired cognition, impaired consciousness, or delirium; 79 (23.7%) of these patients had a head CT scan. Of these 3 groups, delirious patients were more frequently scanned (p < 0.001). Part 2: Of 279 CT scans, 42 (15.0%) were positive for an acute condition (hemorrhage, hematoma, space-occupying lesion, infarct). Of 42 positive scans, 40 (95.1%) were found in the 102 (36.6%) patients with either impaired consciousness or a new focal neurologic finding detected in the ED. CONCLUSIONS: Considerable variability in ED CT scan ordering exists for elder patients with neurologic findings. Impaired consciousness and/or new focal neurologic signs are associated with acute findings on CT scan in elder patients. Acute CT abnormalities are uncommon in elder ED patients with other neurologic findings. Additional prospective evaluation is warranted prior to guideline development for CT scans in this patient population. PMID- 9408424 TI - Proper depth of placement of nasotracheal tubes in adults prior to radiographic confirmation. AB - OBJECTIVE: To determine the optimal initial depth of tube placement in nasotracheal intubation (NTI) of adult patients, measured at the naris, prior to obtaining a chest radiograph (CXR). METHODS: Part 1: A prospective, observational study was performed to compare the initial depth of NTI, measured at the naris, with the observed height of the endotracheal tube (ETT) tip above the carina on the initial CXR. Optimal depths were predicted by gender. Part 2: Results from Part 1 were prospectively validated by measuring the frequency of adequate placement when ETTs were placed to this depth. ETT placement was considered adequate if the tip was at least 2 cm above the carnia and below the larnx on the CXR. RESULTS: Part 1: The mean depth measured at the naris was 27.5 +/- 1.5 cm in women (n = 50) and 27.8 +/- 1.0 cm in men (n = 74). The mean distance of the tip of the ETT to the carina was 3.9 +/- 2.7 cm in women and 6.4 +/- 2.2 cm in men. Initial tube position was adequate in 39 (78%) of the women and 72 (97%) of the men. It was determined that if a depth of 26 cm had been used in the women and 28 cm in the men, 45 (90%) of the women and 70 (95%) of the men would have had adequate tube placement, resulting in statistically significant improvement in the women (p < 0.05; McNemar chi 2). Part 2: These calculated depths (26 and 28 cm) were then prospectively applied in 26 women and 52 men. Twenty-five (96%) of 26 women and 51 (98%) of 52 men had adequate placement, with a mean height above the carina of 4.5 +/- 1.4 cm in women and 5.6 +/- 1.8 cm in men. CONCLUSION: Initial placement of NTI at 26 cm in women and 28 cm in men, measured at the naris, resulted in adequate initial placement for most adult patients. PMID- 9408425 TI - Comparison of routes of flumazenil administration to reverse midazolam-induced respiratory depression in a canine model. AB - OBJECTIVE: To determine whether flumazenil, a drug used to reverse benzodiazepine induced respiratory depression and approved only for i.v. use, is effective by alternative routes. METHODS: A randomized, controlled, nonblinded, crossover canine trial was performed to evaluate reversal of midazolam-induced respiratory depression by flumazenil when administered by alternative routes. Mongrel dogs were sedated with thiopental 19 mg/kg i.v., then tracheally intubated. With the dogs spontaneously breathing, tidal volume, end-tidal CO2, and O2 saturation were observed until a stable baseline was achieved. Incremental doses of midazolam were administered until respiratory depression (30% decline in tidal volume, 10% decrease in O2 saturation, and 15% increase in end-tidal CO2) occurred. Flumazenil was administered by a randomly selected route [0.2 mg followed 1 minute later by 0.3 mg i.v., sublingual (s.l.) or intramuscular (i.m.); or 1 mg followed 1 minute later by 1.5 mg per rectum (PR)]. Time to return to baseline respiratory functions was recorded ("time to reversal"). Each of 10 dogs was studied using all 4 routes of flumazenil administration with a washout period of at least 7 days. An additional dog served as a control (no flumazenil). RESULTS: The control time to reversal was 1,620 seconds. The i.v. route was significantly faster (mean 120 +/- 24.5 sec) than the other 3 routes (p < 0.005). The SL route was the second fastest (mean 262 +/- 94.5 sec), the IM route was the third fastest (mean 310 +/- 133.7 sec) and the PR route was the s;owest (mean 342 +/- 84.4 sec). The SL, IM, and PR routes did not differ significantly from one another. CONCLUSIONS: Flumazenil administered by all 4 routes reversed midazolam induced respiratory depression in a dog model. For the selected dosages used, the i.v. route was significantly faster than all 3 other routes, and SL was the second fastest. PMID- 9408426 TI - Incidence of deep venous thrombosis associated with femoral venous catheterization. AB - OBJECTIVE: To determine in adult medical patients the incidence of deep venous thrombosis (DVT) resulting from femoral venous catheterization (FVC). METHODS: A prospective, observational study was performed at a 420-bed community teaching hospital. Heparin-coated 7-FR cm femoral venous catheters were inserted unilaterally into a femoral vein. Each contralateral leg served as a control site. Age, gender, number of FVC days, DVT risk factors, administration of DVT prophylaxis, and DVT formation and site were tabulated for each patient. Venous duplex sonography was performed bilaterally on each patient within 7 days of femoral venous catheter removal. RESULTS: Catheters were placed in 29 men and 13 women. Femoral DVT was identified by venous duplex sonography in 11 (26.2%) of the FVC legs and none (0%) in the control legs. Posterior tibial and popliteal DVT was identified in both the FVC and control legs of 1 patient. DVT formation at the site of FVC insertion was highly significant (p = 0.005). There were no statistically significant associations with age (p = 0.42), gender (p = 0.73), number of DVT risk factors (p = 0.17), number of FVC days (p = 0.89), or DVT prophylaxis (p = 0.99). CONCLUSION: Placement of femoral catheters for central venous access is associated with a significant incidence of femoral DVT as detected by venous duplex sonography criteria at the site of femoral venous catheter placement. Physicians must be aware of this risk when choosing this vascular access route for adult medical patients. Further studies to assess the relative risk for DVT anf its clinical sequelae when using the femoral vs other central venous catheter routes are indicated. PMID- 9408427 TI - Analysis of the treatment of spontaneous sustained stable ventricular tachycardia. AB - OBJECTIVES: To determine the termination rate of spontaneous sustained stable ventricular tachycardia (SSSVT) as a function of the first and second therapeutic interventions used, and to determine factors associated with successful termination. METHODS: A multihospital, retrospective analysis of the treatment of patients with SSSVT was performed. The setting included 2 urban county hospitals, 2 urban private hospitals, and a Veterans Affairs hospital. Cases were identified by discharge diagnosis and ECG characteristics, and confirmed by electrophysiology study or ECG criteria. RESULTS: There were 40 cases of SSSVT identified. Excluding adenosine, 35 patients were treated with lidocaine as a first intervention. The rate of termination with lidocaine bolus was 17% (6 of 35) (95% CI 7-34%). Regarding the 35 patients initially treated with lidocaine, the odds of termination of SSSVT were 11 times greater in those without a history of previous myocardial infarction (MI) than in those with a history of MI (95% CI 0.96-551). Of the 29 patients who failed initial lidocaine treatment, 23 were treated with a second lidocaine bolus, with a termination rate of 18% (4 of 22) (95% CI 5-40%). Only 2 patients with sustained ventricular tachycardia had a concurrent MI, and the tachycardia was unresponsive to initial lidocaine bolus in both cases. Fifteen patients received adenosine with no tachycardia terminations and no significant adverse effects. CONCLUSIONS: The rate of SSSVT termination with lidocaine was low, particularly in patients with a history of Mi. PMID- 9408428 TI - Adverse outcomes of managed care gatekeeping. AB - OBJECTIVES: To determine whether telephone preauthorization for reimbursement of ED care (medical "gate-keeping") by managed care organizations (MCOs) is associated with adverse outcomes. METHODS: A structured review was performed of case reports solicited during 1994 and 1995 with possible adverse outcomes related to managed care gatekeeping. Gatekeeping was defined as the requirement imposed by an MCO that ED staff contact on-call gatekeepers (i.e., clinical or nonclinical MCO personnel) to request preauthorization for ED treatment (a requirement that such MCOs enforce by refusing payment for the ED care unless preauthorization is obtained). Cases in which gatekeeper denial of preauthorization occurred were sought. Two physicians agreed on patient eligibility and classification criteria, then independently, retrospectively classified case reports identified as MCO ED payment denials into 1 of 4 categories: 1) adverse outcome; 2) patient placed at increased risk of death or disability; 3) "near miss" (emergency physicians prevented adverse outcome by caring for patient despite denial); and 4) none of the above. RESULTS: Of the 143 cases reviewed, 29 reports represented MCO ED payment denial. Of these 29 eligible cases, there were 4 (14%) patients with adverse outcomes, 4 (14%) patients placed at increased risk, and 21 (72%) near misses. All of the 29 cases came from different EDs, representing 9 different states, with the majority from California. Adverse outcomes included respiratory failure from fulminant meningococcemia, hypovolemic syncope from ruptured ectopic pregnancy, hypovolemic arrest from vascular fibroid hemorrhage necessitating emergency hysterectomy, and prolonged postoperative course following ruptured duodenal ulcer. Patients placed at increased risk were diagnosed as having epiglottitis, myocardial infarction, ruptured ectopic pregnancy, and delayed treatment of hip septic arthritis. Near misses included diagnoses of ectopic pregnancy (n = 2), pneumothorax (n = 2), alcohol withdrawal seizures and pancreatitis necessitating intensive care unit admission, appendicitis, bacterial meningitis, cerebrovascular accident, cryptococal meningitis in immuno comprised host, endocarditis, incarerated inguinal hernia, meningocococemia, meninoccocal meningitis, peritonsillar abscess, pneumococcal meningitis, ruptured abdominal aortic aneurysm, shock from gastrointestinal bleeding, small bowel obstruction, schizophrenic crisis resulting in psychiatric hospitalization, suicidal depression resulting in psychiatric hospitalization, and unstable angina. CONCLUSION: Adverse outcomes occur with MCO gatekeeping, Although the present study cannot ascertain whether this is a frequent event or a rare one, the safety of MCO gatekeeping deserves further study. PMID- 9408429 TI - Medically unnecessary pediatric ambulance transports: a medical taxi service? AB - OBJECTIVE: To characterize ambulance utilization in a pediatric population and pediatric emergency physicians' judgement of the medical need for ambulance transport. METHODS: A convenience sample of ambulance transports were studied prospectively during a 5-week period. Exclusion criteria included transfer from another medical facility, study physician not available, need for immediate resuscitation, or trauma team activation. A questionnaire completed by the physician assessed medical need for the ambulance based on chief complaint, general appearance, vital signs, and ambulance run sheet information. A separate questionnaire was administered to the parents regarding reasons for ambulance use and other available means of transportation. Caregivers were contacted by telephone 2-3 days later to determine the mode of transportation home and the clinical outcome. RESULTS: Of 172 eligible patients, 92 (53%) were enrolled. Most (61%; 56/92) transports were considered medically unnecessary. Interestingly, 40% (37/92) of the subjects had no other means of transportation; 86% (32/37) of ambulance transports for this group were judged medically unnecessary. Overall, 86% (79/92) of families had not called their physician. There was no association between having spoken with the physician and medical need for an ambulance. Many (82%; 46/56) Medicaid transports were judged medically unnecessary Overall, follow-up was achieved for 91% (85/92) of the patients. No patient for whom transport was medically unnecessary had a repeat ED visit for the same complaint or required admission. Most patients (74%; 68/92) returned home without any assistance. Among the medically unnecessary transports, 52% (32/60) of the caregivers cited no other means of transportation, yet 34% (11/32) of these patients returned home by private car. CONCLUSIONS: Most pediatric ambulance transports in this sample, which excluded patients requiring immediate resuscitation or trauma team care, were judged to be medically unnecessary. Caregivers often use an ambulance as a convenience or as the only means of transportation. An alternate, less resource-intensive transportation system may be more appropriate for this population. PMID- 9408430 TI - Confidentiality and privacy breaches in a university hospital emergency department. AB - OBJECTIVE: To determine the frequency of visual and auditory confidentiality and privacy breaches in a university ED. METHODS: A prospective, observational study of medical personnel behavior was performed using participant and direct observation techniques. Observations were made in a university tertiary referral and trauma center emergency facility. Observers recorded auditory and visual confidentiality and privacy breaches in various patient care areas during 1-hour periods. Information collected included patient name or room number, complaint/diagnosis, diagnostic tests, past medical history, and personal information. It was then determined whether a clear identification of the patient's name or face and/or an association to his or her clinical course could be made. RESULTS: All members of the health care team committed confidentiality and privacy breaches. Frequency of breaches was dependent on room location and design. Breaches in the triage/waiting area occurred for > 53% of the patients. Breaches near the physician/nursing station ranged from 3 to 24 per hour and 1.5 to 3.4 per patient hour. Other inappropriate comments also were noted. One hundred consecutive patients and family members were interviewed at ED release, with only 2/100 having noticed the status board, although neither could recall any specific details. CONCLUSION: Confidentiality and privacy breaches occur in a university ED by all members of the health care team. The ED architecture and floor plan affect patient confidentiality and privacy. PMID- 9408431 TI - Effectiveness of an organized follow-up system for elder patients released from the emergency department. AB - OBJECTIVES: To determine whether an effective telephone callback system can be successfully implemented in a busy ED and to quantify the benefits that can be obtained related to the follow-up care of elder patients. METHODS: This was a prospective, cohort study conducted at a community teaching hospital during a 6 month period. Consecutive patients > or = 60 years old and released from the ED were selected for telephone follow-up. Calls were made by a research nurse within 72 hours after the patient's ED visit. Follow-up information included current medical status, problems encountered during the ED visit, compliance, and impact of the illness on self-care capabilities. RESULTS: Seventy-nine percent (831/1,048) of the patients selected for telephone follow-up were successfully contacted. The calls lasted an average of 4 +/- 2.5 minutes. Although 94% (778/831) of these patients had a regular physician, 14% failed to make their recommended follow-up arrangements. Compliance was significantly improved when a follow-up physician was contacted during the patient's ED visit. Approximately 96% of the patients were either satisfied or very satisfied with their ED care. However, 13% (109/831) had moderate deterioration in their ability to care for themselves. Of the patients contacted, 333 (40%) required further clarification of their home care instructions, 31 were advised to return to the ED for reevaluation, and 26 were referred to a medical social worker for psychosocial concerns. CONCLUSION: A telephone callback system is a feasible and effective method to improve follow-up care of elder patients released from the ED. PMID- 9408432 TI - Comparison of types of research articles published in emergency medicine and non emergency medicine journals. AB - INTRODUCTION: As the specialty of emergency medicine (EM) matures, its journals should be publishing research of a quality similar to that which appears in other premier journals. OBJECTIVE: To compare the types of original research published in 4 EM vs 3 non-EM journals. METHODS: Retrospective review of all 1995 articles published in Academic Emergency Medicine, American Journal of Emergency Medicine, Annals of Emergency Medicine, Journal of Emergency Medicine, Annals of Internal Medicine, JAMA, and New England Journal of Medicine. Research articles were classified as longitudinal vs cross-sectional, prospective vs retrospective, and interventional vs observational. Other characteristics noted were number of subjects, randomization, blinding, control, and power calculations. Journals were reviewed by 4 investigators who received specific training in research classification, adhering to previously reported criteria for retrospective reviews. Interobserver reliability was independently validated. RESULTS: The authors reviewed 3,524 articles, of which 874 (24.8%) were original research. Compared with research reported in non-EM journals, EM journals contained fewer longitudinal studies (40.5% vs 60.4%, p < 0.0001) and fewer prospective studies (70.8% vs 78.7%, p = 0.008). Fewer EM journals had studies that were blinded (13.7% vs 18.9%, p = 0.047) or controlled (36.3% vs 50.0%, p = 0.003). Studies reported in EM journals had fewer subjects (138 vs 300, p < 0.001). Research reports in EM journals were less likely to have been funded, even after adjustment for the differences in study designs (adjusted odds ratio 7.0, 95% CI 5.1-9.7). CONCLUSION: Significant differences in types of research published in EM and non-EN journals were identified. PMID- 9408433 TI - Implementation of a hypertext-based curriculum for emergency medicine on the World Wide Web. AB - This project reports the publication of a variety of existing curricular resources for emergency medicine on the global Internet in a format that allows hypertext links between related material, timely updates, and end-user feedback. Curricular elements were converted to Hypertext Markup Language with extensive links between related content. The completed document contains instructions for curriculum development, specific curricula for subspecialty areas within a residency, reading lists for subspecialty curricula, banks of images, and board type questions with answers. Users are provided with a mechanism to provide immediate feedback to section editors with suggestions for changes, including new references. Access to all or part of the document can be controlled via passwords, but is potentially available to anyone with an Internet connection and a World Wide Web browser. The document may by viewed on the World Wide Web at: http:@www.brown.edu@Administration@emergency_Medicine@ curr.html. PMID- 9408434 TI - Implications of basic science research for emergency medicine. PMID- 9408435 TI - Septic arthritis of the ankle from Fusobacterium necrophorum. PMID- 9408436 TI - Recurrent pyogenic cholangitis. PMID- 9408437 TI - Documented use of analgesics in the emergency department and upon release of patients with extremity fractures. PMID- 9408438 TI - A model emergency department systemic sedation record. PMID- 9408439 TI - The role of the podocyte in the degeneration of a renal glomerulus. PMID- 9408440 TI - Glomerular cells in the progression of human and experimental nephropathies. PMID- 9408441 TI - Lipids, 3-hydroxy-3-methylglutaryl coenzyme a reductase inhibitors, and progression of renal failure. PMID- 9408442 TI - Optimizing the renal response to ace inhibition: a strategy toward more effective long-term renoprotection. PMID- 9408443 TI - Progression of renal scarring: a balancing act. AB - The progression of renal scarring and fibrosis is a complex process involving intricate interactions between a wide range of pro-scarring and anti-scarring forces and mediators. A better understanding of these events should allow more rational interventions based on their manipulations. Unfortunately, while rapid and considerable progress has been made in experimental animals and in the rat in particular, these so far have had a limited impact on progressive nephropathies in humans. It is as if clinical interventions continue to proceed along different lines, defined decades ago. PMID- 9408444 TI - Some cardiac abnormalities in renal failure. PMID- 9408445 TI - Arterial remodeling and cardiovascular function in end-stage renal disease. PMID- 9408446 TI - The clinical significance of adynamic bone disease in uremia. PMID- 9408447 TI - Peritoneal dialysis: how to improve peritoneal permeability or function. PMID- 9408448 TI - New peritoneal dialysis techniques and their evaluation. PMID- 9408449 TI - Adequacy, nutrition, and outcome in peritoneal dialysis. PMID- 9408450 TI - Epidemiology of toxic nephropathies. PMID- 9408451 TI - Acute renal failure syndromes after bone marrow transplantation. PMID- 9408452 TI - Atherosclerotic renovascular disease in patients with renal failure. PMID- 9408453 TI - Polarized expression of membrane proteins in renal epithelial cells: involvement of specialized transport vesicles and intracellular pathways. AB - The polarized insertion of membrane proteins in epithelial cells is a highly regulated process that involves the coordinated interaction of many subsets of intracellular proteins. Thus, like cog-wheels in a complex machine, the different parts of the trafficking pathway must work together to ensure the correct functioning of an individual cell and of the entire epithelium in which it resides. The individual portions of the trafficking pathway include (1) sorting of membrane proteins into the correct apically or basolaterally targeted vesicles at the level of the trans-Golgi network; (2) delivery of these vesicles to their correct membrane destination by the microtubular (and probably the actin) cytoskeleton; (3) fusion of the vesicles with the appropriate membrane domain via a complex set of fusion proteins. This review has briefly outlined some of these processes and has used examples from the kidney to illustrate their importance to normal renal function. PMID- 9408454 TI - Renal magnesium handling. PMID- 9408455 TI - Mutations in renal ion transporters cause Gitelman's and Bartter's syndromes of inherited hypokalemic alkalosis. PMID- 9408456 TI - Inherited pheochromocytoma. PMID- 9408457 TI - Skin cancers in transplant patients. AB - Skin tumors are frequent in transplant patients, and their potential for progression (locoregional recurrences, metastases) is much greater than in the general population. The viral element with HPV probably represents one of the etiologic factors, although other only partially known factors play a role, including the sun and genetic factors. The high frequency of these skin tumors in transplant patients and their potential for progression require preventive and therapeutic measures: regular examination of the skin, strict advice about protection from sun exposure, excision of any suspect lesion, and treatment of warts that might be conducive to the development of skin cancers. Finally, it must be decided whether immunosuppression should be reduced or stopped during treatment of skin tumors with a high risk of progression. PMID- 9408458 TI - Xenotransplantation: progress toward clinical development. PMID- 9408459 TI - Mechanisms of action of new antiepileptic drugs. PMID- 9408460 TI - Mechanism-specific pathways for new antiepileptic drug discovery. PMID- 9408461 TI - The National Institutes of Health Anticonvulsant Drug Development Program: screening for efficacy. AB - The procedures employed by the ASP provide detailed information pertaining to the anticonvulsant profile of new candidate substances. In addition, the results obtained from tolerance and liver microsomal studies furnish critical information for predicting whether tolerance and/or serious drug-drug interactions are likely to develop following long-term administration of a candidate substance. Finally, in vitro mechanism-of-action studies supply preliminary information regarding the site of action of promising new anticonvulsant drugs. It is anticipated that the testing protocol outlined above will identify safer and mechanistically novel substances to enhance significantly the quality of life of those epilepsy patients still suffering from uncontrolled seizure disorders and/or experiencing significant adverse drug effects. PMID- 9408462 TI - Pharmaceutical industry screening for new antiepileptic drugs. AB - It is clear that in coming years, many interesting new compounds will emerge from these and other strategies for drug discovery. In previous strategies of anticonvulsant discovery, whole-animal models of seizures were utilized from the initial stages; however, with the newer strategies, it will become necessary to select promising drug targets for clinical trials in epilepsy that have well defined molecular targets of action but poorly defined actions in various animal seizure models. The new pharmaceutical discovery technologies of mass screening, cloned drug targets, and combinatorial chemistry will assist in the efficient discovery of novel drug candidates. Incidentally, these techniques also will minimize the use of large numbers of living laboratory animals for anticonvulsant drug screening. PMID- 9408463 TI - Metabolic enzymes and antiepileptic drug interactions. PMID- 9408464 TI - New drugs for persons with epilepsy. AB - Between 30% and 60% of patients with epilepsy have not achieved adequate control with current medications, and side effects are a significant problem. In the past 2 years, three drugs for epilepsy have been approved. At least six more drugs are in the final stages of development, and there is an active "pipeline." None of the new drugs are panceas, but many have special advantages and meet important specific needs. Felbamate, despite a high incidence of aplastic anemia and hepatic failure, remains useful because of its lack of sedative effects and high efficacy. Gabapentin is remarkable for its favorable side effect profile, lack of interactions, and straightforward kinetics. Lamotrigine is also nonsedating and may be especially useful in generalized epilepsies. Topiramate and vigabatrin are both highly effective, although each is associated with a variety of cognitive or psychiatric side effects that may limit utility. Oxcarbazepine shares the efficacy of carbamazepine, with fewer side effects or drug interactions. Zonisamide seems to be effective and cause mild side effects, although the risk for renal stones indicates a need for cautious use. Tiagabine, like gabapentin, is a mild drug with a favorable side effect profile. New forms of old drugs will make for easier administration; fosphenytoin will increase the safety of parenchymal phenytoin use. The best of the new drugs help, at most, 10% of previously uncontrolled patients to become seizure-free. The development of new drugs remains an important need. PMID- 9408465 TI - AED doses: from animals to humans. PMID- 9408466 TI - Assessing pharmacokinetic and pharmacodynamic interactions in clinical trials of antiepileptic drugs. AB - An understanding of the pharmacokinetic and pharmacodynamic properties of a drug is a basic requirement for its clinical use. The investigations of these properties and their timing are fairly clearly defined in the drug development process. Without fundamental knowledge of the pharmacokinetics and pharmacodynamics of a drug, a physician could not use it appropriately, nor would a regulatory agency be likely to approve its use. Information about the interactions of a new antiepileptic drug with other antiepileptic drugs also aids a physician and is required, in varying degrees, by regulatory agencies. The amount of this information that is needed depends, in part, on the class of drug and the population for which the agent is intended. Because antiepileptic drugs are often used as polytherapy and generally are developed first for this use, their interaction potential must be part of the thought process in their development. The correct time to obtain this information, however, is not clearly defined. The analytic methodology to investigate the pharmacokinetic profile of an NCE exists and is fairly sophisticated. This methodology has enabled the development of study designs to investigate pharmacokinetic interactions. Because the plasma concentration of an antiepileptic drug may be increased or decreased as a result of pharmacokinetic interactions with concomitant antiepileptic drugs, it is of great importance to know about the specific interaction potential of an antiepileptic drug early in its development. Recent studies have confirmed the importance of investigating pharmacokinetic interactions in phase I before proceeding into phases II and III. Without this information, study results are often difficult to interpret; with this knowledge, study designs can be modified to minimize the confounding effect. A methodology exists to investigate the pharmacodynamic effects of an antiepileptic drug at receptors in cell cultures and in animal models of seizures; however, no procedure has been established to evaluate the short-term or immediate clinical pharmacodynamic effect of an antiepileptic drug, as has been done for other classes of drugs and other diseases. The clinical effect that is sought in trials with antiepileptic drugs is a reduction in seizures with little toxicity. The methodology to investigate the effect of seizure reduction over time has been used repeatedly with minor variations in the development of all the new antiepileptic drugs. However, no study has evaluated the effect of pharmacodynamic interactions among antiepileptic drugs on seizure reduction. Some studies have purported to show an interaction effect on adverse events, and assumptions are made about pharmacodynamic interactions. Although the information regarding pharmacodynamic interactions is important and existing trial designs could evaluate this, there has been no perceived need to carry out such trials. This information is less accessible than pharmacokinetic interaction information. Moreover, pharmacodynamic interactions, as opposed to pharmacokinetic interactions, are probably unidirectional and lead only to increased effects. Although it would be preferable to have this knowledge, an antiepileptic drug can be used effectively without it; over time, the information about pharmacodynamics will be inferred. Thus, conducting pharmacokinetic interaction studies with antiepileptic drugs early in their development as part of phase I is essential, whereas obtaining pharmacodynamic interaction information can be deferred. PMID- 9408467 TI - Novel methods for studying new antiepileptic drug pharmacology. AB - A number of new antiepileptic drugs act by indirect mechanisms and thus produce effects that may not best be measured by traditional blood studies of the drugs and their metabolites. Study of the indirect action of these drugs on GABA mediated inhibition by microdialysis and nuclear MR spectroscopy has proved more relevant. These new investigative techniques may also prove valuable as compounds affecting glutamate or other excitatory neurotransmitters are developed. PMID- 9408468 TI - The art of antiepileptic trial design. PMID- 9408469 TI - Reanalysis of existing trial data: what can be learned. PMID- 9408470 TI - Monotherapy trials. PMID- 9408471 TI - The impact of foreign data on a new drug application. PMID- 9408472 TI - Issues for women in antiepileptic drug development. PMID- 9408473 TI - Antiepileptic drug trials in children and adolescents: is there a need? PMID- 9408474 TI - Pediatric trials: practical issues. Special populations and trial design. PMID- 9408475 TI - Clinical trials for status epilepticus. PMID- 9408476 TI - Clinical trials for seizure prevention. PMID- 9408477 TI - Alternate endpoints for seizure measurement. AB - The type of seizure measurement selected should be tailored to the needs of the clinical trial. These needs change as the development of the investigational agent moves along. Protocol designers who look toward nontraditional outcome measures might acquire valuable data that could be used to differentiate their drug from other new treatments. Most importantly, as clinical trials evolve into effectiveness studies, clinicians are provided with useful data on which to base selection of a drug of first choice to treat each type of seizure disorder. PMID- 9408478 TI - Alternate endpoints: studies in quality of life and health economics of epilepsy. PMID- 9408480 TI - Routine measurement of new antiepileptic drug concentrations: a critique and a prediction. PMID- 9408481 TI - Cognitive and behavioral assessments in AED trials. PMID- 9408479 TI - Alternate endpoint: EEG assessment of antiepileptic drug efficacy and toxicity. PMID- 9408482 TI - Economic factors in the development of new antiepileptic drugs. AB - The development of new antiepileptic drugs is closely linked to a series of economic factors and perceptions. Industrial interest in such drugs continues to be relatively low, and few companies presently have active discovery programs. The market, however, is large and growing and should encourage more industrial interest in the future. PMID- 9408483 TI - Recent issues and considerations in the regulation of antiepileptic drugs. PMID- 9408484 TI - Ethical and institutional review board issues. AB - IRBs provide an important role in the protection of research subjects/patients. Research investigators have an inherent potential conflict of interest as health care professionals; as physicians, they are dedicated to promoting the welfare of individual patients, whereas as researchers, they seek knowledge that can be generalized and is applicable to persons other than the individual patient under study. The second goal may be in conflict with the first. IRBs have the paramount responsibility of protecting the rights and welfare of human research subjects. Although the IRB system is not perfect, conscientious IRBs reassure the public that the rights and welfare of human subjects are seriously considered by people who do not have a vested interest in the outcome of the research. By exercising their responsibilities, IRBs promote the protection of human subjects. IRB approval provides a significant affirmation of the scientific and ethical qualities of research, and therefore offers important validation to research and research investigators. IRBs, acting in accordance with the guiding principles of the Belmont Report and within the regulatory guidelines of 45.CFR.46, are intended to provide balance between society's interest in advancing scientific knowledge and the mandate to protect the rights and welfare of human subjects. PMID- 9408485 TI - Postmarketing studies. PMID- 9408486 TI - Current management of small-bowel obstruction. AB - In conclusion, most of the recent advances in the management of small bowel obstruction consist of developments in the imaging modalities available to assist in the diagnosis itself, particularly with regard to the distinction between partial and complete obstruction. Unfortunately, little progress has been made to enable physicians to detect early, reversible strangulation, and therefore the surgical management of small-bowel obstruction has changed very little over the past 10 years. Because of the inability to detect reversible ischemia, there is a substantial risk of progression to irreversible ischemia (and an inherent rise in morbidity and mortality) when surgery is delayed for an extended period of time, especially in the setting of suspected complete obstruction. However, almost all patients do benefit from an initial 12 to 24 hours of resuscitation and decompression in cases of complete obstruction; resuscitation and decompression can usually be extended for a longer period of time in those patients with partial obstruction who exhibit no signs of progression (Fig 6). It is encouraging, however, that some advances have been made in understanding the pathophysiology and prevention of adhesion formation. Research efforts in the future should continue to focus on these issues as well as on the development of methods to better recognize early signs of strangulation. PMID- 9408487 TI - Management of minimal breast cancer. PMID- 9408488 TI - Can an enteral diet decrease sepsis after trauma? PMID- 9408489 TI - The role of sentinel lymph node mapping for melanoma. PMID- 9408491 TI - Current surgical management of hepatic cyst disease. PMID- 9408490 TI - Small-diameter portocaval shunt vs. transjugular intrahepatic portosystemic shunt for portal hypertension. PMID- 9408492 TI - Current status of stroma-free hemoglobin. PMID- 9408493 TI - Current management of acute respiratory distress syndrome. PMID- 9408494 TI - Extracorporeal life support for cardiorespiratory failure. PMID- 9408495 TI - The flank vs. the abdominal approach for aortic surgery: the abdominal approach. PMID- 9408496 TI - Flank vs. abdominal approach for abdominal aortic surgery: the flank approach. PMID- 9408497 TI - Extrapleural pneumonectomy for malignant mesothelioma. PMID- 9408498 TI - Reoperation for missed parathyroid adenoma. AB - These results in difficult patients with previously missed parathyroid adenomas demonstrate that a prospective strategy to treat these patients surgically can be used with a high degree of success. This strategy required collaboration among endocrinologists, radiologists, and surgeons. Prospectively, only patients with symptomatic persistent or recurrent primary hyperparathyroidism were included. Operative reports and pathology results from the initial operation and all previous operations were reviewed in detail. Patients with FHH or hypercalcemia from other causes were excluded. State-of-the-art radiologic localization procedures were used to localize the abnormal parathyroid gland. Most patients had only noninvasive procedures, such as the sestamibi scan, but some whose results were equivocal underwent invasive localization, including selective angiography and venous sampling. All patients underwent surgery even if the localization procedures were negative. The operative approach and strategy were dependent on the previous operative result and results of the imaging studies. Operative techniques like IOUS and urinary cAMP determination helped facilitate a more rapid successful outcome. However, in some patients, operative success was only achieved by complete dissection and removal of all tissue that might harbor the adenoma such as a lobe of the thyroid or the thymus. Abnormal parathyroid tissue was cryopreserved for possible subsequent autotransplantation in the event of hypoparathyroidism. The reoperative success rate for missed adenoma was 97%, the highest ever reported with a very acceptable complication rate and no deaths. Postoperative hypoparathyroidism, which was really a complication of the initial previous operations during which normal parathyroid tissue was removed, was treated by autotransplantation of cryopreserved tissue in 12 patients, and 8 functioned perfectly. With attention to details and possible pitfalls, reoperations for missed parathyroid adenomas can be performed safely and effectively. PMID- 9408499 TI - Surgical infections in immunocompromised patients--prevention and treatment. AB - The care of immunocompromised patients is a challenge to all physicians. The variety of opportunistic infections that can develop in these patients underscores the importance of the immune response. The key to success in the prevention and management of infections in these patients lies in knowing what kind of infections to expect and instituting empirical treatment even before a specific diagnosis is made. With the advent of better preventive strategies, the use of prophylactic and preemptive therapies, and the introduction of new immunomodulators, successful outcomes in these interesting and challenging patients may be more easy to achieve in the future. PMID- 9408500 TI - Laparoscopic adrenalectomy. PMID- 9408501 TI - Outcome of surgical therapy for metastatic cancer to the brain. PMID- 9408502 TI - Portal vein involvement in pancreatic cancer: a sign of unresectability? AB - Although the prognosis remains poor, surgical exploration with complete resection provides the only potential cure for patients with adenocarcinoma of the pancreas. However, only 15% to 20% of patients are candidates for resection because of the presence of distant tumor outside the confines of resection or because of locally advanced disease. Even though isolated portal vein involvement has classically been a contraindication for resection, PVR can be performed safely with a low perioperative mortality rate. Besides PV involvement and size, the distribution of histopathologic prognostic factors is no different between patients undergoing PVR and those undergoing standard pancreatic resection. Importantly, overall survival is similar between both groups. Therefore, suspected isolated PV involvement frequently does not preclude operability and, by itself, should not be a contraindication to pancreatic resection. PMID- 9408503 TI - Surgical management of soft-tissue sarcoma. PMID- 9408504 TI - New guidelines address pediatric issues in antiretroviral therapy. PMID- 9408505 TI - Ropinirole approved for Parkinson's disease. PMID- 9408506 TI - New interferon approved for chronic hepatitis C. PMID- 9408507 TI - As patients embrace herbal remedies, dearth of scientific evidence frustrates clinicians. PMID- 9408508 TI - Americans at risk from self-medication, survey reveals. PMID- 9408509 TI - Evidence-Based Practice Centers receive initial assignments. PMID- 9408510 TI - Pharmacist intervention in depression is focus of $3 million research grant. PMID- 9408512 TI - Promoting rational thought about ADHD. PMID- 9408511 TI - Work styles. PMID- 9408513 TI - Oral angiotensin-converting-enzyme inhibitors. AB - The pharmacology, pharmacokinetics, clinical uses, adverse effects, drug interactions, dosage, cost, and therapeutic interchange of oral angiotension converting-enzyme (ACE) inhibitors are reviewed. ACE inhibitors attenuate the formation of angiotension II and may lead to the accumulation of kinins. Although the hypotensive effects of many ACE inhibitors may persist for 24 hours, some patients require more than one dose per day to achieve adequate control. These agents accumulate in patients with renal or hepatic dysfunction, but it is unclear whether dosage adjustments are necessary. ACE inhibitors are effective against mild to moderate hypertension; for severe hypertension, additional anti hypertensive agents may be necessary. Other conditions in which ACE inhibitors have shown efficacy include congestive heart failure, myocardial infarction, left ventricular dysfunction, left ventricular hypertrophy, chronic renal insufficiency, insulin sensitivity, and coronary artery disease. The most common adverse effect is a persistent nonproductive cough. Angioedema, fetal and neonatal morbidity and mortality, acute renal failure, and hyperkalemia may also occur. ACE inhibitors may interact with diuretics, lithium, nonsteroidal anti inflammatory drugs, oral hypoglycemic agents, and some other drugs. ACE inhibitor therapy should be initiated with low doses that may then be slowly adjusted upward. Many of the agents have similar costs for lower and higher dosages. The only significant differences among the ACE inhibitors are the time of onset of hypotensive effects, time to peak effect, and duration of effect. Each formulary should include, at least, captopril and one intermediate-acting and one long acting ACE inhibitor. PMID- 9408514 TI - Influence of piperacillin-tazobactam on pharmacokinetics of gentamicin given once daily. AB - The effect of piperacillin-tazobactam on the pharmacokinetics of gentamicin given once daily was studied. Healthy adult volunteers each received four drug regimens in randomized order: gentamicin 7 mg/kg i.v. once daily (1) by itself, (2) with piperacillin 4 g and tazobactam 0.5 g (both as the sodium salt) every six hours i.v., (3) with piperacillin 4 g and tazobactam 0.5 g i.v. every eight hours, and (4) with piperacillin 8 g and tazobactam 1 g by continuous i.v. infusion over 24 hours. All the gentamicin doses were infused over 30 minutes two hours after piperacillin-tazobactam. Blood samples were drawn before and at the end of each gentamicin infusion and at intervals up to 12 hours after the start of each gentamicin infusion. Samples were assayed for gentamicin concentration by an enzyme-multiplied immunoassay technique and for piperacillin and tazobactam concentrations by high-performance liquid chromatography. Six women and four men completed all four drug regimens. For the gentamicin-alone regimen, the mean +/- S.D. area under the concentration-versus-time curve was 78.06 +/- 10.28 micrograms/mL.hr, the mean +/- S.D. peak concentration (30 minutes after the end of an infusion) was 20.28 +/- 2.54 micrograms/mL, and the mean +/- S.D. half-life was 2.41 +/- 0.24 hours. The values for gentamicin alone did not differ significantly from those for gentamicin in any of the combination regimens. Coadministration of once-daily gentamicin and piperacillin-tazobactam in various i.v. infusion regimens did not affect the pharmacokinetics of once-daily gentamicin. PMID- 9408515 TI - Compatibility of doxorubicin hydrochloride liposome injection with selected other drugs during simulated Y-site administration. AB - The compatibility of doxorubicin hydrochloride liposome injection with selected other drugs during simulated Y-site administration was studied. Five milliliters of doxorubicin hydrochloride liposome injection 0.4 mg/mL in 5% dextrose injection was combined with 5 mL of each of 82 other drugs in 5% dextrose injection or, if necessary to avoid incompatibilities with the diluent, 0.9% sodium chloride injection. The combinations were examined with the unaided eye in fluorescent light and in high-intensity monodirectional light to enhance visualization of small particles and low-level turbidity. The turbidity of each combination was measured as well. Particle sizing and counting were performed on selected combinations. Evaluations were performed initially and at one and four hours. All combinations were stored at room temperature (approximately 23 degrees C). Most of the test drugs were compatible with doxorubicin hydrochloride liposome injection during the four-hour observation period. However, practitioners should be cautious in administering any drug simultaneously with doxorubicin hydrochloride liposome injection until the integrity of the liposomes can be verified. Eighteen drugs exhibited unacceptable increases or decreases in measured turbidity or particulate formation within four hours. During simulated Y site administration, doxorubicin hydrochloride 0.4 mg/mL (as the liposomal injection) in 5% dextrose injection was compatible with 64 of 82 other drugs for four hours at approximately 23 degrees C and was incompatible with 18 of the test drugs. PMID- 9408516 TI - Stability of pyrimethamine in a liquid dosage formulation stored for three months. AB - The stability of pyrimethamine in a liquid dosage formulation stored for up to three months was studies. Commercially available 25-mg pyrimethamine tablets were crushed with a mortar and pestle and mixed with a 1:1 mixture of Simple Syrup, NF, and 1% methylcellulose to yield a suspension with a pyrimethamine concentration of 2 mg/mL. The suspension was poured into 10 amber plastic and 10 amber glass prescription bottles; 5 plastic and 5 glass bottles were stored at 4 degrees C, and the remaining bottles were kept at 25 degrees C. Samples were collected at intervals up to 91 days and tested for pyrimethamine concentration by stability-indicating high-performance liquid chromatography. Pyrimethamine remained stable throughout the three-month study period under all conditions. At 4 degrees C, pyrimethamine concentrations remained above 96% of the initial concentration; at 25 degrees C, pyrimethamine concentrations remained above 91%. No substantial changes in pH were observed. Pyrimethamine was stable for at least 91 days in an oral suspension stored in plastic or glass prescription bottles at 4 or 25 degrees C. PMID- 9408517 TI - Psychopharmacy medication clinic in a managed care women's health setting. PMID- 9408518 TI - Streamlining outpatient drug therapy at a Veterans Affairs medical center. PMID- 9408519 TI - Measuring patient satisfaction with pharmaceutical services. AB - Common conceptualizations of satisfaction are discussed, and different ways of measuring patient satisfaction with pharmaceutical services are suggested. Patient satisfaction is becoming increasingly popular as an indicator of the quality of health care services, including pharmaceutical services. Satisfaction can be conceptualized as a performance evaluation, disconfirmation of expectations, an affect-based assessment, or an equity-based assessment. A satisfaction measure should have a theoretical base on which the measure's validity can be assessed. The measure chosen must fit the context of an overall research process, and the researcher must have a clear idea of what is to be measured. A large pool of items, or questions, for potential inclusion in a patient-satisfaction questionnaire can then be generated. The researcher should develop a format for each item (e.g., Likert scale). The items should be reviewed by experts for relevance and completeness. Questions that will help validate other questions should be included. To assess reliability and validity, the questionnaire should be given to a representative sample of respondents before being refined into its final format. Finally, the researcher must ensure that the patient-satisfaction questionnaire is practical (e.g., in terms of length and complexity). No single standard measure of patient satisfaction is applicable to all pharmacy situations. Researchers should either use an existing measure with demonstrated reliability and validity or develop a new measure by using a systematic process. PMID- 9408520 TI - Human dignity and pharmaceutical care. PMID- 9408521 TI - Determination of cost-effective treatment of acute otitis media from HMO records. PMID- 9408522 TI - More on mechanisms of bacterial resistance. PMID- 9408523 TI - Risk and incidence of asthma attributable to occupational exposure among HMO members. AB - Occupational asthma may account for a significant proportion of adult-onset asthma, but incidence estimates from surveillance of physician reports and workers' compensation data (0.9 to 15/100,000) are lower than expected from community-based cross-sectional studies of asthma patients. We conducted a prospective cohort study of 79,204 health maintenance organization members between the ages of 15 and 55 at risk for asthma. Computerized files, medical records, and telephone interviews were used to identify and characterize asthma cases. Evidence for asthma attributable to occupational exposure was determined from work-related symptoms and workplace exposure. The annual incidence of clinically significant, new-onset asthma was 1.3/1,000, and increased to 3.7/1,000 when cases with reactivation of previously quiescent asthma were included. Criteria for onset of clinically significant asthma attributable to occupational exposure were met by 21% (95% CI 12-32%) of cases giving an incidence of 71/100,000 (95% CI 43-111). Physicians documented asking about work related symptoms in 15% of charts, and recorded suggestive symptoms in three cases, but did not obtain occupational medicine consultation, diagnose occupational asthma, report to the state surveillance program, or bill workers' compensation for any of them. These data suggest that the incidence of asthma attributable to occupational exposures is significantly higher than previously reported, and accounts for a sizable proportion of adult-onset asthma. PMID- 9408524 TI - Antioxidant defense mechanisms in the lung toxicity of tri-n-butyl phosphate. AB - Pneumotoxic effects of the tri-n-butyl phosphate (TBP) are investigated on rats using biological markers in bronchoalveolar lavage fluid (BALF) and studying key antioxidant enzymes in lung homogenate. Each animal from the experimental group received intratracheally 5 microliters TBP (20% v/v in n-dodecane). Six rats from the control and treated groups are sacrificed on post-treatment days 1, 3, 7, 14, and 28. The lactate dehydrogenase activity, the total protein content and the total cell number in BALF are increased mainly on day 1 after the treatment. The activities of superoxide dismutase and catalase are decreased to day 7 and those of glutathione peroxidase and glutathione reductase on day 1 only. The malondialdehyde content is elevated to day 14. It is concluded that TBP causes moderate toxic injury of the lung parenchyma. The depression of the key antioxidant enzymes and the elevated lipid peroxidation are probably important mechanisms of the lung damage. PMID- 9408525 TI - Pulmonary effects of inhaled dust and fumes: exposure-response study in rubber workers. AB - Lung function changes and respiratory symptoms were investigated in a cross sectional study in rubber workers exposed to dust and fumes. To exclude acute pulmonary effects related to "rubber fumes," lung function was measured in curing workers at the start and end of the day shift. Exposure to inhalable dust was measured in all production areas. The results were compared with a reference population from the same geographical region. This study indicates that exposure to "rubber fumes" in curing workers was not related to cross-shift and cross-week decreases in pulmonary function at levels approximately 1 mg/m3 (AM) inhalable dust and 260 micrograms/m3 cyclohexane soluble fraction (CSF). Cross-sectional analyses gave indications for a small loss in pulmonary function in all rubber workers. This decrease in lung function was associated with 10 years of exposure to an average of 2.0 mg/m3 inhalable dust. Our study showed a mean annual decline of 0.08% for the FEV1/FVC ratio and of 10 ml/s for the MMEF. Self-reported chronic respiratory symptoms were not related to dust exposure. PMID- 9408526 TI - Respiratory function and immunological status in cocoa and flour processing workers. AB - Respiratory function and immunological status were studied in 40 cocoa and 53 flour processing workers employed as packers in a confectionery industry and in 65 unexposed control workers in the same industry. A high prevalence of chronic respiratory symptoms was recorded in exposed workers, varying from 5.0% to 30.0% in cocoa workers and from 5.7% to 28.3% in flour workers. Occupational asthma was diagnosed in 2 (5%) of the cocoa workers and in 3 (5.7%) of the flour workers. None of the control workers suffered from occupational asthma. The prevalence of almost all chronic respiratory symptoms was significantly greater in cocoa and flour workers than in control workers. There was also a high prevalence of acute symptoms that developed during the work shift, being highest for cough (cocoa: 57.5%; flour: 50.9%) and eye irritation (cocoa: 50.0%; flour: 54.7%). Significant across-shift reductions of ventilatory capacity were recorded in exposed workers, being largest for flow rates at 50% and the last 25% of the vital capacity on maximum expiratory flow-volume (MEFV) curves (FEF50, FEF75). The prevalence of positive skin tests for cocoa (60.2%) was significantly higher than the prevalence of positive skin tests for flour (25.8%) among the 93 exposed workers (p < 0.05). Control workers had significantly lower prevalences of positive skin tests to cocoa (4.6%) and flour (12.3%) than exposed workers (p < 0.01). Increased total serum IgE levels were found in 17.5% of cocoa and in 18.7% of flour workers; none of the control workers had increased IgE levels. Bronchoprovocation testing demonstrated significant decreases in lung function following inhalation of cocoa dust extract and flour dust in workers with respiratory symptoms and large across-shift reductions in lung function. Dust concentrations in the working environment were higher than those recommended by Croatian standards. These data suggest that workers employed in the processing of cocoa and flour may be at a high risk for the development of allergic sensitization and respiratory impairment. PMID- 9408527 TI - Empirical comparisons of proportional hazards, poisson, and logistic regression modeling of occupational cohort data. AB - This research was conducted to examine the effect of model choice on the epidemiologic interpretation of occupational cohort data. Three multiplicative models commonly employed in the analysis of occupational cohort studies- proportional hazards. Poisson, and logistic regression--were used to analyze data from an historical cohort study of workers exposed to formaldehyde. Samples were taken from this dataset to create a number of predetermined scenarios for comparing the models, varying study size, outcome frequency, strength of risk factors, and follow-up length. The Poisson and proportional hazards models yielded nearly identical relative risk estimates and confidence intervals in all situations except when confounding by age could not be closely controlled in the Poisson analysis. Logistic regression findings were more variable, with risk estimates differing most from the proportional hazards results when there was a common outcome or strong relative risk. The logistic model also provided less precise estimates than the other two. Thus, although logistic was the easiest model to implement, it should be used only in occupational cohort studies when the outcome is rare (5% or less), and the relative risk is less than approximately 2. Even then, the proportional hazards and Poisson models are better choices. Selecting between these two can be based on convenience in most circumstances. PMID- 9408528 TI - Biological monitoring of workers exposed to carbon disulfide. AB - Urinary 2-thiothiazolidine-4-carboxylic acid (TTCA) concentrations and corresponding personal breathing zone carbon disulfide (CS2) air concentrations were measured for worker populations at a Tennessee rubber product facility and a Virginia viscose rayon plant. At the rubber product facility, all of the 19 workers had urinary TTCA levels less than the limit of detection (LOD) of about 0.03 mg/L, equivalent to less than 0.5 ppm of CS2 in air. At the viscose rayon plant, five of six workers, although wearing half-mask cartridge respirators, showed increased urinary TTCA during the workshift. The cutters and spinners had the largest increases in urinary TTCA concentrations; they also had the highest breathing zone exposures to CS2. The TTCA concentrations for three cutters and spinners did not return to normal preshift levels of < 0.3 mg/g creatinine before the start of the next shift. The arithmetic mean respirator workplace protection factor (WPF) was 7.0 +/- 2.2. Increase in urinary TTCA concentration during the workshift and postshift urinary TTCA concentration reflected CS2 air concentration equally well. In conjunction with air monitoring results, urinary TTCA concentrations allow determination of the WPF afforded workers wearing respirators and identify workers not adhering to safety rules and good work practices. Workers at risk of adverse health effects from overexposure to CS2 for any reason may thus be identified. PMID- 9408529 TI - Urine vanadium concentrations in workers overhauling an oil-fired boiler. AB - Since fuel oil ash contains vanadium (V), the measurement of urinary levels of V may provide a biological marker in workers exposed to fuel oil ash. The usefulness of urine V samples as a biological monitoring tool ultimately depends on determining the appropriate time of sampling relative to when exposure occurs. Twenty boilermakers were studied during the overhaul of a large oil-fired boiler. A total of 117 urine samples were collected, 65 start-of-shift (S-O-S) and 52 end of-shift (E-O-S) samples. Air V exposures were estimated with personal sampling devices and work history diaries. Air V concentrations ranged from 0.36 to 32.19 micrograms V/m3, with a mean +/- SD of 19.1 +/- 10.7, and a median of 18.5. On the first day of work on the overhaul, the V urine levels at the E-O-S (mean +/- SD were 1.53 +/- 0.53, median was 1.52 mg V/g creatinine) were significantly higher than those at the S-O-S (0.87 +/- 0.32, median was 0.83), P = 0.004. However, the V concentrations of the S-O-S urine samples on the last Monday of the study were not significantly different from the S-O-S urine levels on the previous Saturday, a time interval of about 38 hr between the end of exposure and sample collection. The Spearman correlation coefficient (r) between the S-O-S urine V and the workplace concentration of V dust during the previous day was r = 0.35. In summary, the results suggest a rapid initial clearance of V (elevating the E-O-S V concentration on the first day of work relative to the S-O-S concentration), followed by a slow clearance that is not complete 38 hr after the end of exposure, as evidenced by the Monday morning urine V concentrations. The Spearman correlations suggest that the S-O-S urine is preferred to the E-O-S urine for across-shift biological monitoring of V exposure. PMID- 9408530 TI - An updated mortality study of workers at a petroleum refinery in Beaumont, Texas. AB - The mortality experience of 7,119 workers who were employed at a Beaumont, Texas, refinery for at least 1 year between 1945 and 1987 was investigated. Mortality analyses based on standardized mortality ratios (SMRs) and 95% confidence intervals (95% CI) showed overall mortality was significantly lower than expected compared with the U.S. general population (SMR = 82, 95% CI = 79-86). Total cancer mortality was also lower than expected (SMR = 92, 95% CI = 84-100). Significant mortality deficits from several malignant and nonmalignant diseases were reported. A significant mortality increase in the broad category of lymphatic and hematopoietic cancers was found (SMR = 133, 95% CI = 103-170). This increase was attributed to a nonsignificant elevation in leukemia of all cell types combined (SMR = 139, 95% CI = 92-201) and a borderline significant increase in other lymphatic tissue cancer (SMR = 158, 95% CI = 101-235). The elevation in leukemia was confined to workers hired before 1950. Furthermore, the leukemia excess was shown to have peaked during the 1960s, with mortality no longer elevated post-1980. Analyses of cell type-specific leukemias showed a similar temporal pattern for acute myeloid leukemia (AML) which was not significantly elevated (SMR = 136, 95% CI = 59-268). Mortality from other leukemia cell types was similar to or lower than expected. Mortality from non-Hodgkin's lymphoma (NHL) (SMR = 140, 95% CI = 88-211) and multiple myeloma (MM) (SMR = 121, 95% CI = 55-230) were increased, but neither was statistically significant nor likely to be related to refinery employment. No death from asbestosis was reported, and mortality from mesothelioma and pulmonary fibrosis was lower than expected. Lung cancer mortality for the overall cohort was similar to expected. For the overall cohort, analyses by duration of employment and time since first employment showed no evidence of any trends for increasing cause-specific mortality. Separate analyses of male workers employed in operator jobs showed mortality patterns that were more favorable than those of the total cohort. Maintenance craftworkers showed statistically significant elevations in mortality for prostate cancer (SMR = 145, 95% CI = 107-194), leukemia (SMR = 179, 95% CI = 111-273), and other lymphatic tissue cancer (SMR = 233, 95% CI = 138-368). Detailed analyses indicated that, among maintenance craftworkers, mortality was elevated for AML, NHL, and MM, but none was significant. Furthermore, no upward trend by duration of maintenance jobs was observed. A small increase of lung cancer was observed among maintenance craftworkers (SMR = 120, 95% CI = 99-145), which was borderline significant. No relationship between lung cancer and duration of maintenance employment was found. In contrast, a deficit of pulmonary fibrosis was reported among maintenance craftworkers (SMR = 62, 95% CI = 17-159). These findings are discussed in conjunction with results from other refinery studies, and the limitations of the study are discussed. PMID- 9408531 TI - Non-hodgkin's lymphoma and agricultural use of the insecticide lindane. AB - Data from population-based case-control studies of non-Hodgkin's lymphoma among white men from Kansas, Nebraska, Iowa, and Minnesota were pooled to evaluate potential risks from environmental exposures in more detail, while controlling for potential confounding factors. These data provided the opportunity to evaluate the risk of non-Hodgkin's lymphoma from potential exposures to lindane, a pesticide that causes cancer in laboratory animals and has been associated with human cancer in a few epidemiologic investigations. This pooled data set includes 987 individuals with non-Hodgkin's lymphoma and 2,895 population-based controls. Information was obtained by telephone or in person interviews, which included detailed questions on farm practices and agricultural use of chemicals. Logistic regression was used to calculate odds ratios (ORs) adjusted for age, state of residence, and subject or proxy interviews. Reported use of lindane significantly increased the risk of non-Hodgkin's's lymphoma by 50%. Some use characteristics were suggestive of an association. ORs were greater among persons who first used the pesticide 20 years before diagnosis (OR = 1.7) than more recently (OR = 1.3), among those who reported more frequent use (OR = 2.0 for use 5 or more days per year versus 1.6 for fewer than five days per year), and from use on crops (OR = 1.9), rather than from use on animals (OR = 1.3), although these differences were not statistically significant. On the other hand, ORs were lower when based on direct interviews (OR = 1.3) than on data from proxy respondents (OR = 2.1) and adjustment for potential confounding by use of 2,4-D and diazinon reduced the ORs associated with lindane use from 1.5 to 1.2 and 1.3, respectively. Lindane does not appear to be a major etiologic factor in the development of non-Hodgkin's's lymphoma, although a small role cannot be ruled out. PMID- 9408532 TI - Cancer among greenhouse owners and their relatives: results of a pilot study. PMID- 9408533 TI - A cross-sectional study on asbestos workers carried out in Italy in 1940: a forgotten study. PMID- 9408534 TI - RE main asbestos type in pleural mesothelioma. PMID- 9408535 TI - Further examination of employment in the chemical industry: follow-up case control study of hematopoietic and lymphoid neoplasms using a next consecutive death control group. PMID- 9408536 TI - Bioarchaeological research in the Mariana Islands of the western Pacific: an overview. PMID- 9408537 TI - A biocultural perspective on Marianas prehistory: recent trends in bioarchaeological research. AB - Fifteen years ago, the biohistory of Micronesia was still a blank slate relative to other regions of the Pacific. Since 1980, however, the Mariana Islands, one of the largest island chains in Micronesia, have been the focus of intensive archaeological investigation and human remains have been ubiquitous components of the archaeological assemblages recovered from the islands of Guam, Rota, Tinian, and Saipan. These investigations have provided us with a wealth of new data that will contribute substantially to our understanding of population adaptation to island ecosystems in this part of the Pacific. Much of the recent bioarchaeological research in the Marianas is the product of archaeological mitigation rather than directed research. Consequently, many of our research efforts have been articulated with the needs of cultural resource management (CRM) where research designs focus on several general problem areas: 1) subsistence adaptation with emphasis on the contribution of marine vs. terrestrial resources and the role of pelagic, or deep-ocean resources in the marine component of the diet; 2) regional (upland vs. coastal; interisland) and temporal variation in subsistence/settlement; and 3) geomorphologic variation in coastal sediments, particularly as influenced by storm events. PMID- 9408538 TI - Skeletal biology of Apurguan: a precontact Chamorro site on Guam. AB - The human skeletal remains of a minimum of 152 individuals from the precontact Latte Period (AD 1000-1521) on Guam, Mariana Islands, are described. The sample, recovered at Apurguan, in the Tamuning District, is one of the largest series of well-provenienced Chamorro skeletal remains to be analyzed in recent years. The size and systematic nature of this database are a major contribution to the human biology of the region. Paleodemographic characteristics, dental and skeletal morphology, and paleopathology are presented, along with a limited examination of sex differences in frequencies of nonmetric variation. The mortuary sample, consisting of 51 subadults and 101 adults, exhibits underrepresentation of females, highest subadult mortality between 2 and 10 years, and an adult average age-at-death of 43.5 years. Cranial and infracranial indices and nonmetric variation are consistent with the Chamorro's Southeast Asian origins. There are few statistically significant sex differences in nonmetric variation which suggests close genetic affinity. The frequency of dental pathology overall is low, reflecting a well-balanced, varied diet, and consistent with preagricultural subsistence; however statistically significant sex differences suggest the influence of differential cultural behaviors or resource access. Paleopathological observations include healed fractures (more common in males), little advanced osteoarthritis, evidence for gouty arthritis, and treponemal disease (yaws). One individual, a young adult male, was interred with 10 human bone spear points in situ. Twenty percent of the primary burials exhibit evidence of postdepositional removal of selected skeletal elements for cultural purposes such as keepsakes or raw material. PMID- 9408539 TI - An assessment of health and disease in the prehistoric inhabitants of the Mariana Islands. AB - Using a variety of skeletal and dental stress indicators, an assessment of the health and disease of the indigenous inhabitants of the Mariana Islands, the Chamorro, is made. The major hypothesis to be tested is that the Chamorro were relatively healthy and that deviations from the expected, as well as inter-island variation, may reflect environmental, ecological, and cultural differences. The major skeletal series surveyed include sites on Guam (N = 247 individuals), Rota (N = 14), Tinian (N = 20), and Saipan (N = 102). The majority of the specimens are from the transitional pre-Latte (AD 1-1000) and Latte (AD 1000-1521) periods. These data derive primarily from unpublished osteological reports. The indicators of health and disease surveyed include mortality and paleodemographic data, stature, dental paleopathology, cribra orbitalia, limb bone fractures, degenerative osteoarthritis, and infectious disease (including treponemal infection). Where appropriate, tests of significance are calculated to determine the presence of any patterning in the differences observed within and between the skeletal series. Information recorded in prehistoric Hawaiians provides a standard for external comparisons. Several of the larger skeletal series surveyed have paleodemographic features that are consistent with long-term cemetery populations. Females and subadults are typically underrepresented. Most subadult deaths occur in the 2-5 year age interval. Life expectancy at birth ranges from 26.4 to 33.7 years. A healthy fertility rate is indicated for these series. The prehistoric inhabitants of the Mariana Islands were relatively tall, exceeding living Chamorros measured in the early part of the present century. The greater prevalence of developmental defects in the enamel suggests that the Chamorro were exposed to more stress than prehistoric Hawaiians. The low frequency of cribra orbitalia further indicates iron deficiency anemia was not a problem. There are generally low frequencies of dental pathology in the remains from the Mariana Islands. Betel-nut staining is relatively common in all series which may help to explain the relatively low prevalence of dental pathology. Healed limb bone fractures are rare in these skeletal series; the frequency and patterns of fractures suggest accidental injury as the main cause. Greater physical demands involving the lower back region are indicated by a high frequency of spondylolysis, or stress fracture in the lumbar vertebrae in the Chamorro. Likewise, advanced degenerative bone changes, while of low occurrence, are significantly greater in the Chamorro than Hawaiians. The prevalence of skeletal and dental indicators of stress was generally higher in the smaller islands of the Mariansas chain (e.g., Rota), islands with fewer resources to buffer environmental catastrophe. Bony changes suggestive of treponemal (probably yaws) disease are common in most of these Marianas Islands skeletal series. PMID- 9408540 TI - Stable isotopic analysis of human diet in the Marianas Archipelago, western Pacific. AB - Proportions of marine vs. terrestrial resources in prehistoric human diets in the southern Mariana Islands (Guam, Rota, Saipan), Micronesia, have been estimated by analysis of stable isotope ratios of carbon and nitrogen in bone collagen and of carbon in apatite. The isotopic composition of marine and terrestrial food resources from the Marianas have also been determined. Experimental evidence shows that collagen carbon isotopes mainly reflect those of dietary protein sources and thus overestimate the contribution of marine animal foods. Marine protein consumption apparently ranges from approximately 20% to approximately 50% on these islands. Experiments also demonstrate the carbon isotope ratio of bone apatite carbonate accurately reflects that of the whole diet. Carbonate carbon isotope data suggest some individuals consumed significant amounts of 13C enriched (C4) plants or seaweeds. Sugar cane is an indigenous C4 crop and seaweeds are eaten throughout the Pacific, but they have not been considered by archaeologists to have been prehistoric dietary staples. Apatite carbon isotope analysis has apparently identified previously unrecognized prehistoric dietary adaptations in the Mariana Islands, but this must be confirmed by archaeobotanical evidence. PMID- 9408541 TI - Subadult stress, morbidity, and longevity in Latte Period populations on Guam, Mariana Islands. AB - The frequency and age distribution of linear enamel hypoplasia (LEH) in the dentition of 293 individuals from Latte Period sites (AD 800-1521) on Guam, Mariana Islands, are examined in this study. Individuals dying as subadults (before age 16) and as young adults (ages 16-21) have more frequent LEHs than those who survived to middle or late adulthood, documenting a relationship between LEH-causing stress events and reduced life expectancy. The age distributions of cribra orbitalia and skeletal infection in children who died by age 10 exhibit striking similarities to the etiological age patterns of LEH in children, and those with skeletal infection have more frequent hypoplasias than children without infection. The comorbidity of systemic stress and infection in children, and their impact on life expectancy, are interpreted in the biocultural context of high population density in the large coastal villages of the late prehistoric period in the Marianas. PMID- 9408542 TI - Cultural alteration of human teeth in the Mariana Islands. AB - Evidence of cultural dental modification in a precontact (pre-1521) skeletal sample from the Academy of Our Lady of Guam gymnasium site in Agana, Guam, is documented. Two of the four individuals recovered at the Academy Gym site exhibit modification of the maxillary teeth. One individual displays vertical incising of a single tooth, and the other exhibits horizontal abrading of the anterior teeth which may be a purposeful or an incidental alteration. Although deliberate alteration of the dentition, including tooth extraction, notching, filing, and drilling, has been documented in human groups worldwide, little has been written about these cultural practices in the Mariana Islands. Examination of the available literature on precontact human remains from the region reveals at least three patterns of dental incising and similar cases of dental abrasion. While the origins of these practices are not known, the presence and style of these cultural alterations may be sex-specific, cosmetic in nature, or an indication of status in a ranked society. Alternatively, they may signify membership in a particular group or lineage, or mark a rite of passage. Because the comparative samples are limited in number and small, and the provenience of many of the skeletons is obscure, temporal variation cannot be ruled out. PMID- 9408543 TI - Spondylolysis in prehistoric human remains from Guam and its possible etiology. AB - This study reports the findings of complete bilateral separation of the neural arch (spondylolysis) in 176 inhumations from the Hyatt Site, Tumon Bay, which is located on the west side of the island of Guam. Skeletons were excavated and analyzed by the Paul H. Rosendahl Inc. (PHRI) team in 1989-1990. The inhumations were associated with the pre-European Latte Period (circa 1,200-1,521 A.D.). This period was characterized by the use of large stone pillars, called latte sets, for the construction of houses. Of the 176 individuals, only 38 adult skeletons had complete spines, and 21% (8/38) of these had evidence of spondylolysis in their lumbar vertebrae, particularly in L-5. The age of the eight individuals range from 30 to 50 years. No children were found with spondylolysis. Of the males 29.4% (5/17) had spondylolysis, as did 14.3% (3/21) of the females. However, the difference between the sexes was not statistically significant. Though the sample is small, it is suggested that the high incidence of lumbar spondylolysis found in these ancient Chamorros was related to lower back traumatic events. The transport of latte stones, involving hyperextension and torque of the lower back, while dragging the stones, probably contributed to the development of microfractures in the spine and subsequent spondylolysis. If this hypothesis is correct, then both males and females appear to have been participants in an organized community labor force. It is predicted that similar frequencies of spondylolysis will be found at other Latte sites. PMID- 9408544 TI - Cranial variation in prehistoric human skeletal remains from the Marianas. AB - Nonmetric cranial variation and facial flatness of the Pacific and circum-Pacific populations are investigated. The peoples of the Marianas, eastern Polynesia and Hawaii form a cluster and show affinities in terms of nonmetric cranial variation with the Southeast and East Asians rather than with the Jomon-Ainu, a view which is widely supported by others. Facial flatness analysis also indicates that Polynesians have different patterns of facial prominence as compared with the Jomon-Ainu. These results increase the difficulty of accepting the Jomon-Pacific cluster proposed by Brace and his coworkers. Although genetic and nonmetric cranial variation reveal relatively close relationships, the Mariana skeletons are markedly different in facial flatness and limb bone morphology from those of Polynesians. PMID- 9408545 TI - Craniofacial affinities of Mariana Islanders and circum-Pacific peoples. AB - Metric craniofacial variation was studied in a number of skeletal samples that originated from the Mariana Islands and circum-Pacific regions. The broad comparisons including East/Southeast Asians, Polynesians, Melanesians, and Australians confirm the relationships between Mariana Islanders and East/Southeast Asians on the one hand and Polynesians on the other hand. A transformation of Melanesians into western Micronesians is not supported. The result of the principal component analysis indicates that the cranial morphological pattern of Mariana people shares the intermediate characteristics between those of typical East/Southeast Asians and several groups falling as outliers to more predominant Asian populations. PMID- 9408546 TI - Cardiovascular disease in the NHANES III. PMID- 9408547 TI - Epidemiology and the health care revolution. AB - PURPOSE: Revolutions in the health care marketplace, with their implied dependence upon the population perspective, have renewed interest in epidemiologic methods and their application. METHODS: As a way of organizing questions, data, and answers, epidemiology is once again the core scientific discipline for a multidimensional paradigm shift that emphasizes five facets of health care. First, the health of enrolled populations, rather than merely treatment of sick individuals presenting for care. Second, assessment of the "time, place, and person" of disease burden--who is healthy, who is not, and what it will take to maximize their health. Third, the causative factors for disease and the factors or characteristics that promote health. Fourth, the development of evidence-based clinical guidelines for clinicians and consumers alike. Fifth, evaluation of the impact providers are having on the health of populations (outcomes), individually and collectively. RESULTS: Thoughtful consideration of all of the above factors depends upon epidemiologic data and insights brought to a firm and conclusive end point capable of supporting policy. CONCLUSIONS: Credibility will require that epidemiologists (and the symbiotic media) exercise far greater restraint in drawing definitive conclusions and speculations from often starting but meager data. PMID- 9408548 TI - Occupational epidemiology in China comes of age. PMID- 9408549 TI - Risk factors and early detection of lung cancer in a cohort of Chinese tin miners. AB - PURPOSE: To examine risk factors and establish a biologic specimen and data bank for the study of early markers of lung cancer. METHODS: We designed a dynamic cohort using an ongoing lung cancer screening program among radon- and arsenic exposed tin miners in Yunnan China. Through the first four years of the study, 8,346 miners aged 40 years and older with over 10 years of occupational exposure have been enrolled, risk factors have been assessed, annual sputum and chest radiographs have been obtained, and numerous biologic specimens have been collected. RESULTS: A total of 243 new lung cancer cases have been identified through 1995. Radon and arsenic exposures are the predominant risk factors, but lung cancer risk is also associated with chronic bronchitis and silicosis, as well as a number of exposure to tobacco smoke, including early age of first use, duration, and cumulative exposure. Tumor and sputum samples are being examined for early markers of lung cancer. CONCLUSION: A cohort of occupationally-exposed tin miners with an extensive biologic specimen repository has been successfully established to simultaneously study the etiology and early detection of lung cancer. PMID- 9408550 TI - DSM-IV alcohol dependence and drinking in the U.S. population: a risk analysis. AB - PURPOSE: This paper examines the relationship between alcohol dependence according to the criteria found in the 4th edition of the Diagnostic and Statistical Manual (DSM-IV) of the American Psychiatric Association and drinking in the U.S. general population. METHODS: The data set under analysis is the 1988 National Health Interview Survey, which interviewed a probability sample of 22,102 adult drinkers in the U.S. household population. The response rate was 86%. RESULTS: Results indicate that there is a linear relationship between DSM-IV dependence and the mean number of drinks consumed per day, or the number of days drinking five or more glasses of alcohol in the past 12 months. Respondents who reported consuming five or more drinks in a day have about six times more chances of being dependent than respondents who did not report such pattern of drinking. Older drinkers are less at risk than younger drinkers. CONCLUSIONS: There is a risk of alcohol dependence at relatively low volumes of consumption. The risk increases gradually with the volume of consumption. An added and higher risk exists when drinkers engage in a pattern of consumption involving the ingestion of five or more drinks per day. PMID- 9408551 TI - Self-esteem and adiposity in black and white girls: the NHLBI Growth and Health Study. AB - PURPOSE: Obesity is assumed to have a negative impact on self-esteem because of the associated social stigmatization in Western society. Studies of the psychological effect of obesity in children are inconclusive and limited, particularly pertaining to minority populations. Most studies have assessed global rather than domain-specific measures of self-esteem and hence, may have lacked specificity to detect impairment of certain aspects of self-esteem most closely associated with obesity. The purpose of this study is to examine the effect of adiposity and other environmental factors on measures of perceived competence and self-adequacy in 2205 black and white girls aged 9-10 years. METHODS: Domain-specific measures of self-esteem were studied by race and degree of adiposity, using Harter's "Self-Perception Profile for Children". Three Harter scales deemed more relevant to obesity (social acceptance (SA), physical appearance (PA), and global self-worth (GSW)) were selected for univariate and multivariate linear regression models to examine relationships between self esteem level and adiposity (measured by the sum of triceps, subscapular, and suprailiac skinfolds (SSF)), race, pubertal maturation, and parental education. The relationship between adiposity and Harter scores was further examined with LOESS curves and also by comparing the mean scores of each quintile of SSF by race, as well as inter-quintile differences within race. RESULTS: Adiposity in general impacted negatively on the scores of all three selected Harter scales. There was also racial variation in the relationship between the scores and adiposity, with the magnitude of the effect somewhat less in black girls. White girls exhibited a significant inverse relationship between SSF and SA scores while, in striking contrast, there was no variation in scores in black girls across all ranges of adiposity. Although there was a significant inverse relationship between adiposity and PA and GSW in both groups, the slope was steeper in white girls, particularly at higher ranges of SSF. Non-linearity in the relationship between SSF and the scores was seen in SA and PA scales. CONCLUSIONS: The present study demonstrated a significant negative association between adiposity and the level of self-esteem in girls as young as 9 to 10 years. There were also intriguing racial differences in the selected domains of esteem. These results may help better understand cultural differences regarding the psychological impact of obesity and could be used to formulate appropriate strategies for public health policy. PMID- 9408553 TI - Risk of suicide attempts after benzodiazepine and/or antidepressant use. AB - PURPOSE: Recent publications imply the existence of associations between psychotropic drugs use and risk of suicide. Some studies have measured the tissue level of these drugs in suicide deaths, while others compared toxicity indices, defined as number of suicide deaths per million prescriptions for individual antidepressants. Few of these studies used unexposed controls. The objective of this cohort study was to evaluate suicide attempts in subjects recently exposed to benzodiazepines and/or antidepressants, as compared to unexposed controls. METHODS: A population of 225,796 persons with prescriptions for benzodiazepines were selected from the Saskatchewan Health Data Bases. Controls consisted of 97,862 individuals who did not receive benzodiazepines. RESULTS: Stratifying the populations into antidepressant users and non-antidepressant users indicated that nonantidepressant users had statistically significant associations between suicide attempts and benzodiazepine use (odds ratio (OR) = 6.2), antipsychotic use, (OR = 2.6), and a history of past treatment for drug/alcohol abuse (OR = 13.4). Antidepressant users showed a statistically significant relation only with past treatment for drug/alcohol use (OR = 5.8). It is argued that the large OR for antidepressant use is due to confounding by indication. If so, the concept of toxicity index is misleading and should not be used. CONCLUSIONS: The association between benzodiazepine use and attempted suicide is especially high for nonantidepressant users, for the young, and for males. Whether this relationship is causal or not, physicians should be aware of the high potential for suicide attempts when prescribing benzodiazepines for patients in these high risk groups. PMID- 9408552 TI - Associations of oral contraceptive use with serum lipids and lipoproteins in young women: the Bogalusa Heart Study. AB - PURPOSE: Cross-sectional and longitudinal associations of serum lipids and lipoproteins with oral contraceptive (OC) use were examined among white and black women aged 18-27 years in 1985-86 and 1988-1991 in the Bogalusa Heart Study, a study of cardiovascular disease in a Southern community. METHODS: Analyses of covariance. RESULTS: In 1985-1986, white OC users had significantly (p < 0.05) higher adjusted mean total and low density lipoprotein (LDL) cholesterols, and lower high density lipoprotein (HDL) cholesterol compared with nonusers; black OC users had higher triglycerides and LDL cholesterol, and lower HDL cholesterol. In 1988-1991, white OC users had higher total cholesterol, triglycerides, and LDL cholesterol, while black OC users had higher triglycerides. OC use was unrelated to mean HDL cholesterol levels in 1988-1991; however, a lower percentage of white OC users than nonusers in 1988-1991 had HDL cholesterol levels < 35 mg/dl. Longitudinally, white OC nonusers at baseline who used OCs at follow-up had significant increases from baseline levels in total cholesterol, triglycerides, and very low density lipoprotein (VLDL) and LDL cholesterols; black women showed an increase only in LDL cholesterol. White women who stopped using OCs by follow up had a decrease in VLDL and LDL cholesterols, and an increase in HDL cholesterol. White OC users at both exams also had a significant increase in HDL cholesterol, whereas women who began using OCs by follow-up did not. CONCLUSIONS: The unfavorable lipid profile associated with OC use was not apparent upon discontinued use. Lack of an adverse effect of OC use on HDL cholesterol at follow-up may be the result of changing formulations, and requires further examination. PMID- 9408554 TI - Pharmacokinetics of selected antibacterial agents. PMID- 9408555 TI - An in-flight investigation of the efficacy of dextroamphetamine for sustaining helicopter pilot performance. AB - A promising countermeasure for fatigue in sustained aviation operations is stimulant administration. However, well-controlled, aviation-relevant studies of the efficacy of medications such as Dexedrine are virtually nonexistent. In this investigation, flight performance, mood, and alertness were evaluated in 10 UH-60 pilots during sleep deprivation periods under Dexedrine or placebo. Relative to placebo, Dexedrine improved flight performance during straight-and-levels, climbs, descents, right turns, and a left-descending turn, with tendencies toward better performance during the left turns and the instrument landing system approach. Dexedrine markedly reduced subjective feelings of fatigue, confusion, and depression while increasing feelings of vigor. Central nervous system arousal was enhanced by Dexedrine relative to placebo. No significant side effects occurred, although Dexedrine was associated with mild asymptomatic increases in heart rate and BP. Thus, Dexedrine appears effective for the short-term sustainment of aviator performance during sustained operations. However, future work should investigate the efficacy of stimulants for longer-term use (e.g., more than 40 h of continuous wakefulness). PMID- 9408556 TI - Validity of using non-pilot subjects to represent pilots in a sustained acceleration environment. AB - BACKGROUND: A preliminary study determined the similarities between the personality of military pilots (transport and fighter) and centrifuge subjects using the Edwards Personal Preference Schedule (EPPS). Past, similar personality studies have shown differences between military fighter vs. transport pilots, and general population vs. male and female general aviators. To use subjects in lieu of pilots in the centrifuge, they must represent the pilot characteristics of interest, for both ethical and scientific reasons. With the increase in measuring performance metrics (e.g., reaction time, tracking tasks, missile evasion) during centrifuge testing, any factor effecting performance must be explored. It is unknown whether personality effects performance. METHODS: Cluster analysis of 36 pilot and subject personality tests consisted of the Partitioning Around Medoids (PAM) program by Leonard Kaufman and Peter Rousseeuw (10) and Ward's method/K MEANS clustering (CSS:STATISTICA). RESULTS: The clusters generated by the 36 pilots and subjects did not match the Retzlaff and Gibertini (21) clusters. Two clusters were preferred over three, and while the values of the personality variables Dominance, Exhibition, and Aggression (DOM, EXH, AGG) were similar, the pilot membership did not coincide. Subjects had basically the same cluster characteristics as pilots and did not alter the pilot cluster composition characteristics when clustered together. Females did not appear to differ from the males in the cluster analysis. Clustering did not differentiate between fighter and transport pilots using the chosen variables. CONCLUSION: These preliminary results support the hypothesis that there are no major differences in personality between fighter pilots, transport pilots, or centrifuge subjects using the EPPS. PMID- 9408557 TI - Stress in Air Force aviators facing the combat environment. AB - BACKGROUND: This paper evaluates the effect of stress on four squadrons of United States Air Force aviators in tactical high performance aircraft deployed for combat operations compared with U.S. based aircrew using the Beck Depression Inventory (BDI) as the evaluating instrument. METHODS: This is a retrospective cross-sectional study consisting of 42 aviators in deployed squadrons stationed overseas and involved in a contingency mission, and 15 subjects stationed in the U.S. and not exposed to combat conditions. Each subject was administered the test instrument, which was completed in privacy and with complete anonymity. RESULTS: The hypotheses of interest were: a) the proportion of individuals in the population of fighter aircrew who would report excessive stress is 0; and b) no significant differences would exist in the proportion of individuals with excessive stress in the various squadrons. Using statistical methodology, these hypotheses were rejected. CONCLUSION: It is concluded that more studies in each given circumstance are necessary. PMID- 9408558 TI - Content analysis of the crew communication with external communicants under prolonged isolation. AB - BACKGROUND: The communication between space crews and outside monitoring personnel in Mission Control gives us valuable information about crewmember psychological health. Studying the formal parameters of similar communication during ground-based space simulation missions helps us understand the psychological climate of confined groups over time, and this information can be applied to long-term space missions. HYPOTHESES: We hypothesized that psychological closing and information filtration will be observed in a confined crew as the mission progresses and will differ by the type of communication (e.g., audio vs. computer-generated) with the outside monitors in the simulated Mission Control (MC). In addition, communication patterns will vary with the specific group of people on-duty outside. METHOD: Audio and computer communications between space simulation crews and MC teams were analyzed during two isolation studies lasting 135 and 90 d at the Institute for Biomedical Problems in Moscow. A number of temporal, content and quantitative parameters of verbal activity were examined. RESULTS: The total intensity of communication dropped after month 1 of the missions. The Commander had the highest intensity of communication. There was evidence for the presence of psychological closing and information filtration in the crews over time, with some variation based on type of communication. Communication patterns differed with different outside teams. CONCLUSIONS: The communication between confined groups and outside monitoring personnel is affected by psychological closing and information filtration and by the make-up of the teams that comprise the monitoring groups. PMID- 9408559 TI - Cardiovascular effects of a sustained -Gz force in the horizontal position. AB - INTRODUCTION: The purpose of this investigation was to study the effect of a sustained -Gz force applied in the horizontal position on the cardiovascular system and the distribution of body fluids. METHODS: The study was conducted on four men who were submitted to two protocols. The principal difference between the two protocols was that in Protocol A the subjects remained supine, whereas in protocol B the subjects were submitted to a small -Gz force (0.38 G) for a relatively prolonged period while remaining supine. The -Gz force was generated by a short arm centrifuge (AGS or artificial gravity simulator) which has four beds radially oriented on a circular platform and allows variation of the Gz force by controlling the angular velocity. The cardiovascular and fluid distribution responses were sampled approximately every 2 min during the whole duration of the experiment (205 min). The variables recorded were thoracic impedance (Tz), stroke volume (SV), heart rate (HR), first derivative of the thoracic impedance change (dz.dt-1), and systolic (SBP) and diastolic (DBP) arterial BPs. RESULTS: Changes in these variables within each protocol and across protocols were analyzed statistically by group comparisons. This analysis showed that changing from the standing to the supine position causes an increase in SV and a decrease in HR and SBP and DBP. Adding a -Gz force produces the same qualitative effects but causes a greater increase in SV and a greater decrease in systolic and diastolic BPs, even though the -Gz force was only a fraction of 1 G. CONCLUSION: Changing position from vertical to horizontal position and the application of a -G force produce similar cardiovascular and body fluid effects but these effects are amplified by -Gz. An attempt to find an explanation to those changes has led us to hypothesize that a combination of factors such as central venous pressure and sequestering fluids in some part of the thorax must play an important role in the characteristics of those changes. Further research on the physiological effects of prolonged -Gz may be relevant to studies in simulated microgravity. PMID- 9408560 TI - Low gravity and inertial effects on the growth of E. coli and B. subtilis in semi solid media. AB - BACKGROUND: Several published experimental results have shown that cultures of suspended bacteria exhibit increased growth in the spaceflight environment. HYPOTHESIS AND METHODS: To test whether these differences were due to fluid mechanics and not cellular effects, E. coli and B. subtilis were grown on agar cultures under static, agitated, and rotated conditions in the laboratory, and under low-gravity conditions on four Space Shuttle flights. Growth experiments were terminated with glutaraldehyde, and individual cells were counted after quantitative elution from the agar. RESULTS: The spaceflight results, in conjunction with static, rotation, and agitation experiments indicate that E. coli and B. subtilis cultures on agar, unlike their suspension grown counterparts, do not experience heightened final cell concentration when the inertial environment is changed. CONCLUSIONS: This finding points to fluid dynamics and extracellular transport phenomena and not cellular dynamics as the most likely cause of previously reported increases in bacterial growth in microgravity. PMID- 9408561 TI - Sympatho-vagal responses in humans to thermoneutral head-out water immersion. AB - To clarify the role of autonomic nervous functions in cardiovascular adaptation to microgravity, heart rate variability (HRV) and blood pressure variability (BPV) were evaluated during thermoneutral head-out water immersion (HOI) of eight healthy young subjects 23 to 31 yr of age. The very low-frequency (VLF; 0.00-0.04 Hz) component of BPV tended to increase during HOI, whereas the low-frequency (LF; 0.04-0.15 Hz) component of BPV and the ratio of LF power to high-frequency (HF; 0.15-0.40 Hz) component (LF/HF ratio) of HRV decreased. The HF component of HRV increased in all the subjects during immersion up to the shoulder. Concomitantly, we found a decrease in heart rate and increases in stroke volume and cardiac output with no significant changes in BP and respiration rate during HOI. These results suggest that both vasomotor and cardiac sympathetic activities are suppressed and that the parasympathetic (vagal) activity is enhanced during HOI. PMID- 9408562 TI - Effects of fore-and-aft, lateral and vertical whole-body vibration on a head positioning task. AB - BACKGROUND: The performance of tasks in which the head must be positioned close to objects in a moving vehicle may be impeded by the presence of vibration. HYPOTHESES: It was hypothesized that the extent to which a head positioning task would be impeded by whole-body vibration would depend on the frequency, direction and waveform of the vibration and the posture of the body. METHOD: There were 12 subjects who participated in a laboratory experiment in which they judged the difficulty of looking through a pair of sights while exposed to low frequency vibration. We investigated 4 variables: vibration axis (fore-and-aft, lateral, vertical), vibration frequency (11 frequencies in the range 0.5 to 5.0 Hz), vibration waveform (sinusoidal vibration, one-third octave bands of random vibration), seating condition (wearing a 4-point harness, sitting without back support). RESULTS: We found that all variables affected the perceived task difficulty. Frequencies of horizontal vibration in the range 1 to 4 Hz caused most difficulty. Task difficulty was greatest with random vibration, especially with low frequency vibration in the horizontal axes. The wearing of a 4-point harness greatly reduced the perceived task difficulty during exposure to low frequency fore-and-aft vibration but increased task difficulty with higher frequencies of lateral vibration. CONCLUSIONS: Increased motion predictability and the provision of suitable support to the upper body (e.g., a harness, back support, front support) can reduce the difficulty of head positioning tasks during exposure to some types of oscillatory motion. PMID- 9408563 TI - High altitude effects on human taste intensity and hedonics. AB - BACKGROUND: The study was conducted on human volunteers taken to 3500 m altitude for a period of 3 wk. METHODS: Subjects rated four compounds representing sweet, salty, sour and bitter taste, and the hedonic matrix in terms of taste threshold, taste intensity, and taste hedonicity were recorded using category scale. Blood sugar levels were estimated weekly. RESULTS: An increase in the taste thresholds for glucose and sodium chloride was shown while quinine sulphate and citric acid thresholds recorded a decrease. The taste intensity ratings showed a linear relationship with increasing logarithmic molar concentrations of each solution, as compared with taste hedonicity which showed an inverted 'U' type function. The blood picture did not reveal any change in the blood sugar level. All the parameters recorded at high altitude (HA) showed a tendency to return to basal values after reinduction to sea level. CONCLUSION: The study suggests that HA hypoxic stress brings about changes in the hedonic responses, primarily an increased palatibility for sweetness; we speculate that the mechanism may be anorexia-linked nutritional stress. PMID- 9408564 TI - The influence of hyperbaric oxygenation on leukocyte viability and surface protein expression. AB - Hyperbaric oxygenation was studied as a potential inducer of cell growth and differentiation in promyelocytic leukemic HL60 cells. We studied changes in HL60 cells exposed to hyperbaric oxygen, oxygen, or carbon dioxide for 72 h. The proliferation rate and viability of cells in the hyperbaric oxygenation groups were significantly lower (p < 0.05) than for the controls. While CD13 and CD38 were expressed less following hyperbaric treatment, CD11b, CD14, and CD16 showed an increase following hyperbaric treatment, and CD10, CD15, and HLADR showed no change. These results support previous studies which demonstrate the role of oxygen tension in the regulation of cell cycle and protein expression. PMID- 9408565 TI - Passenger doctors in civil airliners--obligations, duties and standards of care. AB - Airlines frequently rely on passenger doctors to assist with in-flight medical emergencies, but the legal implications of such actions vary between nations. While no examples of actions taken against treating physicians for alleged negligence in such emergencies were found, examples of recourse to litigation against individual airlines where the advice of passenger doctors was deemed to be incorrect are cited. Legislation regarding the obligation to treat patients involved in in-flight medical emergencies, the duty and standard of care once treatment, including any history-taking and examination, has commenced, and the requirement for consent to any examination or treatment that may be required is discussed. Particular emphasis is placed on comparing and contrasting the approach of the law in the United Kingdom and United States with the law in countries using Civil Law systems. The paper concludes with a hypothetical example of the kind of difficulties that may practically be encountered and a list of recommendations for physicians attending medical emergencies in civil airliners. PMID- 9408566 TI - Case report: obstructive sleep apnea--an air safety risk. AB - Aviation safety reports indicate that many incidents are related to fatigue. Obstructive sleep apnea (OSA) is characterized by irregular snoring with repeated apnea episodes during sleep and excessive daytime sleepiness. Deprived of sleep, patients suffer from daytime sleepiness and involuntary sleep attacks. The prevalence of OSA among adult men is more than one percent, 0.5% in women. Predisposed are men aged 40-65 yr. Many patients, including pilots, are unaware of their sleeping disturbance and the symptoms are not easily recognized. Therefore, this condition may not be discovered during a regular health examination. However, this condition can be effectively treated. In our opinion, pilots suffering from OSA do not necessarily have to lose their certificate. Diagnosis and treatment can be conducted, followed by regular check-ups. We suggest that questions about sleep be included in pilots' health examinations. PMID- 9408567 TI - How to measure human adaptation in extreme environments: the case of Antarctic wintering-over. AB - BACKGROUND: Whether wintering-over causes cognitive and sensory impairment and associated adaptation problems is still a subject of debate. Conflicting results reported in the literature may be due to the difference in the experimental tasks. Moreover, interpersonal variability can hide the effects. HYPOTHESIS: Performance on cognitive and psychomotor tasks can be an indicator of adaptation problems. METHODS: Subjects were 10 scientists and 6 technicians (both clinically normal) who wintered-over for a year in the Antarctic. A subset of tasks of the AGARD battery of Standardized Tests for Research with Environmental Stressors (SB) was completed eight times during the isolation. To assess adaptation, the adaptability questionnaire (AQ) was completed by the physician before every performance test. RESULTS: SB showed a sensitive period of adaptation at mid winter and at the end of the isolation, but the performance in the different tasks did not deteriorate exactly at the same time. No systematic relationship between AQ notations and SB performances was noted. However, three subjects showed significant positive correlations: r > 0.8, p < 0.05 (i.e., low adaptation = low performance). CONCLUSION: Some clinically normal individuals can experience adaptation problems, and SB tasks can be an indicator of these problems when comparisons are done at an individual level. PMID- 9408571 TI - A unique flight experience: earthquake relief support. PMID- 9408568 TI - East meets West: a comparison of eastern block/western aeromedical practices. AB - Under the auspices of the European Command (EUCOM) Military-to-Military Exchange Program, the authors participated in 13 trips to visit aeromedical facilities of nine Eastern European nations (Albania, Belarus, Bulgaria, Czech Republic, Hungary, Lithuania, Poland, Romania, and Slovakia). In addition, eight of these Eastern European nations visited United States Air Force (USAF) aeromedical facilities. This article highlights the similarities and differences noted between the USAF and Eastern Europe in the practice of aerospace medicine. Flight surgeons from both Eastern Europe and the USAF address issues such as physiologic stresses of flight (acceleration, hypoxia, etc.) and lifestyle stresses (rest, diet, alcohol, cigarettes, etc.). Eastern European Flight Surgeons do not regularly fly. The Eastern European approach to medical standards and screening for aviation applicants is much stricter and more comprehensive than ours. Several of the nations visited had active research programs at their central aeromedical institute emphasizing aircrew selection and retention standards. With the exception of the Czech Republic, Eastern European nations did not routinely grant waivers for chronic medical conditions such as hypertension in aircrew. Soviet-built aircraft had many unique features such as an outside-in attitude indicator and an auto-recovery system. PMID- 9408572 TI - Possible aircrew intoxication caused by accidental release of RainBoe: a case report. PMID- 9408573 TI - Organic solvents and exacerbation of M.S. PMID- 9408574 TI - Finite element analysis of anterior cervical spine interbody fusion. AB - The present study investigated the external and the internal biomechanical responses of anterior cervical discectomy coupled with fusion. Five different types of interbody fusion materials were used: titanium core, titanium cage, tricortical iliac crest, tantalum core, and tantalum cage. Two different types of surgical procedures were analyzed: Smith-Robinson and Bailey-Badgley. A validated three-dimensional anatomically accurate finite element model of the human cervical spine was used in the study. The finite element model was exercised in compression, flexion, extension, and lateral bending for the intact case and for the two surgical procedures with five implant materials. The external response in terms of the stiffness and angular rotation, and the internal response in terms of the disc and the vertebral stresses were determined. The Smith-Robinson technique resulted in the highest increase in external response under all modes of loading for all implant materials. In contrast, the Bailey-Badgley technique produced a higher increase in the disc and the vertebral body stresses than the Smith-Robinson technique. As experimental human cadaver tests can only determine the external response of the non-fused spine simulating immediate post-operative structure, the present finite element studies assist in the understanding of biomechanics of interbody fusion by delineating the changes in the extrinsic and intrinsic characteristics of the cervical spine components due to surgery. PMID- 9408575 TI - Modeling the tensile behavior of human Achilles tendon. AB - Uniaxial quasi-static tensile stress, sigma versus strain, epsilon, data were obtained from 29 cadaveric Achilles tendons (donor ages: 36 to 100 years), at a strain rate of either 10 or 100%/s. These results were then used in modeling the elastic component of the tensile deformational behavior of this tissue. Two approaches were taken. In the first, it was shown that the following constitutive relation provided an excellent fit to the elastic section of the sigma-epsilon curve, sigma = C epsilon exp[D epsilon + F epsilon 2], with C, D and F being material constants, whose values for the present dataset were found to be C = 2.00 +/- 0.99, D = 0.089 +/- 0.087 and F = -0.0047 +/- 0.0095. The values of these coefficients were not statistically significantly affected by either donor age or test strain rate. In the second approach, the value of the modulus of elasticity of a filamentary polymer matrix composite material was computed as a function of various combinations of values of the modulus of elasticity of the fiber, the modulus of elasticity of the matrix, and angle of orientation of the principal material axes with respect to the reference coordinate axes (theta) for a fiber volume fraction of 0.6 and a material Poisson's ratio of 0.4. By comparing these results with the experimentally-obtained values of the tangent modulus of elasticity of the tendons (defined as the slope of the linear section of the post-toe zone in the sigma-epsilon plot), and assuming that the tendon may be idealized as a filamentary polymer matrix composite material, the suggestion is made that the winding angle of the fibers (collagen fibrils) in the tendon (taken to be equal to theta) is about 6 degrees. PMID- 9408576 TI - Reinforcement of PMMA bone cement with a continuous wire coil--a canine femur study. AB - The hoop and axial strains present due to loading after femoral stem implantation in canine femurs, implanted with and without a wire coil surrounding the distal tip were investigated. One stem served as the control without wire coil while the other was experimental reinforced with a wire coil. Both stems were implanted by identical methods. Ideally, the wire coil should serve to reduce the hoop and axial strains present in the distal tip of the arthroplasty. The strains were measured using 90 degrees rosette strain gages. Though the coil's position was altered slightly during implantation usable results were still obtained. At a maximum load of 44.5 N there were 32 and 19% reduction in the hoop and axial strains for the reinforced stem and the control, respectively. This experiment presents a striking difference between the control and reinforced hip arthroplasties. Equally important is that this study confirms the trends in hoop and axial strain behavior demonstrated in other works utilizing a wire coil reinforcement scheme. The simple method of applying a continuous wire coil may help to reduce the loosening of femoral stem of total hip arthroplasty by reducing strains at the tip of the stem due to the strengthening of the cement mantle. PMID- 9408577 TI - New titanium alloys for biomaterials: a study of mechanical and corrosion properties and cytotoxicity. AB - Three new titanium alloys with Zr, Nb, Ta, Pd and In as alloying elements were developed and compared with currently used implant metals, namely, pure Ti and Ti 6Al-4V alloy, in terms of mechanical and corrosion properties, and cytotoxicity. New alloys showed comparable mechanical properties with that of the Ti-6Al-4V alloy, but increased corrosion potential, somewhat decreased breakdown potential and increased corrosion rate. There were no significant differences in cell growth on the surface of the various metal specimens, indicating that the cells cannot differentiate between the passivated surfaces of the various Ti metals. PMID- 9408578 TI - Bulge ductility of several occlusal contact measuring paper-based and plastic based sheets. AB - Articulating paper/film is a sheet strip that is coated with ink- or dye containing wax and is used for marking occlusal contacts and monitoring the results. New types of occlusal film systems have been developed recently, which are capable of being computer-interfaced to identify the occlusal contact points and digitize the occlusal force. The mechanical ductility and thickness of these occlusal sheets constitute some of their important properties. The objective of this study was to evaluate seven different occlusal sheets and compare their bulge ductility and thickness. A custom-designed photo-sensing bulge tester was used. Three paper-based sheets (BAP, BET and SFA), two plastic-based sheets (ACF and AOS), and advanced systems (TSS and FDP) were tested. The specimen size was 20 mm square. Fifteen samples were tested for each material. The sheet film thickness with coated layers was as follows: BAP, 62 microns; BET, 46 microns; SFA, 133 microns; ACF, 23 microns; AOS, 14 microns; TSS, 134 microns; and FDP, 82 microns. Ductility (standard deviation) was as follows: BAP, 2.10 (0.0060)%; BET, 2.14 (0.008)%; SFA, 5.19 (0.57)%; ACF, 8.68 (0.05)%; AOS, 16.26 (0.41)%; TSS, 16.26 (0.41)%; and FDP, 6.37 (0.09)%. One-way ANOVA analysis indicated that (1) there in no statistical difference between BAP and BET (p > 0.001), (2) there is a statistical difference (p < 0.001) among all the rest of the tested occlusal sheets, and (3) bulge ductility appears to be linearly related to film thickness, its correlation depending upon types of base materials. PMID- 9408579 TI - Fracture analysis of cast pure Ti and Ti-6Al-4V alloy for dental use. AB - The fracture behavior of cast two types of pure Ti and Ti-6Al-4V alloy was understood by acoustic emission (AE) analysis during a fracture toughness test. Specimens for test were cast by the lost wax method using a specially designed Ti casting machine of pressure-different method for dental use. A fatigue crack was inserted from the machined notch tip into the body of a specimen in the range of 0.45-0.55 a/W. Acoustic emission signals released during the fracture toughness test were detected by two sensors attached to both ends of the specimen. Then the signals were recorded and analysed by the PAC 3000/3104 system. From the early stage of the fracture toughness test, AE signals started to be released in all types of specimens tested. A reaction layer with the investment materials of about 50-100 microns was thought to be the result of the AE release from an early stage of the fracture toughness test. A microfracture behavior of the cast pure Ti and Ti alloys was proposed based on the results obtained from the AE releasing pattern and fracture surface findings. PMID- 9408580 TI - Locking strength of Morse tapers used for modular segmental bone defect replacement prostheses. AB - Mechanical testing has been performed to characterize the locking strength of Morse taper locks used for reconstruction of large bone defects. Taper joint pairs were locked with a series of compressive loads increasing from 500 to 3500 N. Following each load application the taper locks were distracted with either an axial load or a torsional load. Additional tapers were loaded with 2 million cycles of axial compression or 2 million cycles of cantilever bending combined with axial compression, followed by axial distraction. The torsional and axial distraction loads increased linearly with the compressive load. Compared to a single compressive load application, cyclic axial loading had little influence on the joint strength, while a combination of axial loading and bending increased the joint strength. Based on these results, in vivo loading should increase the locking strength of Morse taper locks used for bone defect reconstruction. PMID- 9408581 TI - Differential staining of two subpopulations of Purkinje neurons in rat cerebellum with acid dyes. AB - We present a new method that stains differently two subpopulations of Purkinje cells in the adult rat. Deparaffinized sections of cerebella, fixed by perfusion with buffered glutaraldehyde or Bouin's fluid were stained with 0.5% light green in 50% ethanol (10-30 min). The excess dye was removed with saturated aqueous picric acid (10-30 min). At this point some Purkinje cells appeared as lightly stained neurons, while others were strongly stained. Slides were immersed in 0.5% aqueous acid fuchsin for approximately 1 min until the lightly stained neurons acquired a red color. Following immersion in 1% phosphotungstic acid, slides were rapidly dehydrated in ethanol, passed to xylene and mounted in Canada balsam. Two subpopulations of Purkinje cells differing in their protein content in somata and proximal dendrites stained differentially by this method. They occurred in all coronal and sagittal sections and in patches or stripes. Their relative proportion varied from lobule to lobule. A second staining method used potassium permanganate as the sole staining reagent. The staining reagent can be used on sections previously stained with the acid dyes. Purkinje cells appeared as subsets of brownish to deep brown stained neurons, the latter ones corresponding to green stained cells in the dichromic method. The results obtained indicated that the subpopulations reflect real differences among individual neurons and are not artifacts. The technique holds promise for identifying and localizing sub sets of Purkinje cells differing in their protein content under normal and experimental conditions and for their further characterization by combined staining and histochemical procedures. PMID- 9408582 TI - Complete staining of nerve fiber and myoneural junctions with acetylthiocholine and silver. AB - We describe a combined stain for simultaneous demonstration of the preterminal axons and cholinesterase activity at myoneural junctions of mammalian muscles. This technique employs acetylthiocholine iodide as the substrate for cholinesterase activity and silver nitrate impregnation of preterminal axons. The procedure is rapid, simple and uses fresh muscles. Intramuscular nerves, preterminal axons and myoneural junctions are stained simultaneously brown or black with minimal background staining of connective tissue and muscle fibers. PMID- 9408583 TI - Fine needle tissue aspiration for accurate localization of histological specimens from complex structures. AB - A procedure is presented for exact, detailed comparison of light and electron microscopic analyses of tissues with complex architecture. Earlier techniques require one to make drawings of tissue pieces to be analyzed by electron microscopy to permit rough localization of the origin of the tissue pieces. Specifically, exact analysis of fetal cartilage and bone is hampered by the complicated arrangement of both tissue components, severely limiting the assessment of electron microscopic analyses. The advantage of the technique described here is that it allows precise localization of the tissue sample in the original tissue area. Punches 1 min in diameter were obtained from femora and coxae with a syringe and embedded for light and electron microscopy. The remaining tissue with its exactly defined punctures is prepared for standard histology. Human fetal cartilage and bone tissue were used to demonstrate this technique, but this procedure may be used for other kinds of tissues. PMID- 9408584 TI - An improved method for chromosome counting in maize. AB - An improved method for counting chromosomes in maize (Zea mays L.) is presented. Application of cold treatment (5C, 24 hr), heat treatment (42 C, 5 min) and a second cold treatment (5C, 24 hr) to root tips before fixation increased the number of condensed and dispersed countable metaphase chromosome figures. Fixed root tips were prepared by the enzymatic maceration-air drying method and preparations were stained with acetic orcein. Under favorable conditions, one preparation with 50-100 countable chromosome figures could be obtained in diploid maize using this method. Conditions affecting the dispersion of the chromosomes are described. This technique is especially useful for determining the somatic chromosome number in triploid and tetraploid maize lines. PMID- 9408585 TI - DiOC6(3): a useful dye for staining the endoplasmic reticulum. AB - The present review discusses the fluorescent organelle probe, DiOC6(3), with reference to its structure, chemistry, availability, spectral properties, labeling procedures, vital staining characteristics, and major applications in cellular and molecular biology. The specificity of dye for endoplasmic reticulum is summarized. We examine the simplicity and advantages of the fluorescent dye system for evaluating structure and function of endoplasmic reticulum. Other significant uses of the dye are also discussed. PMID- 9408586 TI - Apyrase, ascorbic acid and aprotinin ameliorate the storage lesion in pelleted platelet preparations. AB - To evaluate the effect of apyrase, ascorbic acid and aprotinin (AAA) in preventing platelet activation during storage, 12 sets of platelet concentrates (PCs), were treated with AAA and evaluated at days 1, 3, and 5 utilizing platelet functional and morphological assays. Platelets treated with AAA demonstrated significantly enhanced response to ADP-induced platelet aggregation, higher morphology scores, and evaluated ATP levels compared to control samples after 5 days of storage. Similarly, platelet specimens treated with AAA had significantly reduced PF4 secretion and P-selectin expression compared to controls. Finally, Western blots of aggregated platelets at day 5 demonstrated that AAA-treated PCs continue to express the platelet membrane GPIb whereas specimens from control PCs do not. These results show that PCs treated with AAA have reduced platelet activation and enhanced functional platelet activity. PMID- 9408587 TI - The use of dimethylsulfoxide for fixation of yeasts for electron microscopy. AB - Conventional methods of chemical fixation are often inadequate for preserving yeast ultrastructure. The thick cell wall severely limits penetration of fixatives rendering poor detail of the cell wall, membranes, and overall anatomy. Dimethylsulfoxide (DMSO) enhances penetration of chemicals and has been added to fixatives to improve cell preservation. At high concentrations (5 to 50%), however, it affects ultrastructure unpredictably. We found that adding 0.1% DMSO to fixatives greatly improved retention of yeast ultrastructure. Candida albicans, C. glabrata and Aspergillus fumigatus were fixed for 3 hr in 3% paraformaldehyde, 1% glutaraldehyde, 1 mM MgCl2, 1 mM CaCl2, 0.1% DMSO in 0.1 M sodium cacodylate buffer followed by 1% OsO4, 1% K2Cr2O7, 0.85% NaCl, 0.1% DMSO in the same buffer. Thin epoxy sections were post-stained in uranyl acetate and lead citrate. The multilayered character of the cell wall was distinct and well structured. Addition of ruthenium red or alcian blue to the fixatives further enhanced the outer fibrillar layer. The plasma membrane was contiguous and tightly adjacent to the inner mannoprotein layer of the cell wall. The cytoplasm was well preserved and the overall preservation of the yeast ultrastructure was significantly improved. PMID- 9408588 TI - Commercial formalin substitutes for histopathology. AB - We compared the performance of six commercial fixatives proposed to be formalin substitutes with the performance of buffered formalin, Clarke's ethanol-acetic acid, and ethanol, using rat liver, small intestine, and kidney. We investigated the rate of penetration, mode of fixation, extent of protein and structural immobilization, quality of histology and cellular structure following routine dehydration and paraffin embedding, and performance as a fixative for immunohistochemistry. Furthermore, we evaluated the effects of the various fixatives on ultrastructure. Only buffered formalin performed equally well on all tissues tested. While several of the commercial fixatives appeared to preserve liver tissue at 200x, the preservation of kidney, intestinal villi, and smooth muscle was unacceptable. Histological distortion, cell shrinkage and vacuolization were prominent when the substitutes or ethanol were used. In contrast, these artifacts were found occasionally and to a minor degree when buffered formalin or Clarke's fixative were used. Immunohistochemistry demonstrated a total loss of low molecular weight antigens for all fixatives except buffered formalin. The best immunostaining was obtained by combining formalin fixation with antigen retrieval. We conclude that none of the proposed commercial substitutes for buffered formalin are adequate for critical histology or histopathology. PMID- 9408589 TI - Electrical status epilepticus during sleep (ESES). Different clinical syndromes: towards a unifying view? AB - 'Electrical status epilepticus during sleep' (ESES) is a typical childhood process of generalization of paroxysmal activity. Notwithstanding a number of intermediate forms, three syndromes included in the 1989 ILAE classification can be considered as prototypes: the 'continuous spike-waves during sleep' (CSWS syndrome), the 'acquired aphasia with convulsive disorder in children' (L-Kl syndrome) and the 'benign epilepsy of childhood with rolandic spikes' (BECRS), which can be considered as the benign end of the spectrum. The pathognomonic clinical and EEG features of these conditions are described. They can probably be considered, in a unifying view, to be based on a common pathogenetic factor. They are associated with neuropsychological and/or mental disturbances with differences probably due to the idiopathic or symptomatic origin of the underlying epileptic condition, the cortical area of the primary focal paroxysmal activity, the patient's age and the severity and duration of the paroxysmal dysfunction. Possible hypotheses on the physiopathogeneses of ESES and correlated neuropsychological disorders are summarized. Short cycles (3-4 weeks) of relatively high daily doses of diazepam (DZP) (0.5 mg/kg body weight) following a rectal DZP bolus of 1 mg/kg b.w. seem to be effective in the majority of ESES conditions. The somewhat underestimated problem of neuropsychological disorders correlated with ESES in BECRS is also considered. PMID- 9408590 TI - Tissue culture study on neuronal migration in the rat cerebral cortex: effects of low dose radiation. AB - In order to elucidate the molecular mechanisms underlying neuronal migration in the developing rat cerebral cortex, a novel primary tissue culture system in which neuronal migration can be evaluated was developed. Using this culture system, through autoradiographic studies we demonstrated the migration of [3H]thymidine labeled cells, and also revealed that this neuronal migration was delayed by such low dose radiation as 10 cGy. In addition, an immunohistochemical study revealed that the neural cell adhesion molecule (N-CAM), was undetectable in the matrix cell layer. When anti-N-CAM monoclonal antibodies were added to the tissue culture system, the neuronal migration delay was comparable with that observed in the case of 20 cGy radiation. These results suggest that N-CAM is related to neuronal migration in the rat cerebrum and that the neuronal migration delay evoked by low dose radiation might be caused by disorder of N-CAM present in the matrix cells. PMID- 9408591 TI - Vigabatrin as add-on therapy in children and adolescents with refractory epilepsy: an open trial. AB - Seventy-seven children and adolescents with drug-resistant epilepsies received vigabatrin as add-on therapy for a median of 18 months (range 4-36 months) at a dose of 50 mg/kg/day divided in two doses; patients with spasms were given a maximum dose of 100 mg/kg/day. In 23 patients (29.9%), seizure frequency decreased by 50-100% and in 12 patients (15.6%) by 25-50%. The number of seizures remained unchanged in 34 patients (44.1%) and increased in seven (9.1%). Vigabatrin was most effective in cryptogenic and symptomatic partial seizures (39% and 43%, respectively), and in infantile spasms (25%). Adverse events occurred in 20 patients (26%), though they were generally mild and transient, suggesting that vigabatrin is well tolerated. PMID- 9408592 TI - Molecular and pathological studies in Charcot-Marie-Tooth disease 1A. AB - We analyzed a 1.5-Mb duplication of the p11.2-12 region of chromosome 17, including the PMP-22 gene (CMT1A duplication), seven families with Charcot-Marie Tooth disease type I (CMT I) and six sporadic patients with suspected CMT I by Southern blot analysis. In order to detect the CMT 1A duplication, probe pVAW409R3a, probe PMP-22 cDNA and reference probe SF85 were used for Southern hybridization. In six out of seven families with CMT I, CMT1A duplication was identified. One of six sporadic CMT patients had CMT1A duplication. The probe pVAW4O9R3a was more informative than PMP-22 cDNA and SF85 for detecting CMT1A duplication. In pathological study of biopsied sural nerve, thickened myelin sheath was observed in some myelinated fibers in patients with CMT1A duplication. PMID- 9408594 TI - Benign myoclonic epilepsy: long-term follow-up of 11 new cases. AB - The authors report a long-term follow-up of 11 new subjects with benign myoclonic epilepsy. There were some unusual clinical features such as the need for dual therapy in 45.5% of subjects, and the presence of non-epileptic myoclonus in 54.5%, neither of which influenced the prognosis. Neuropsychological and behavioral evolution was less favorable in 45.5% of patients (mental retardation, school learning problems, attention deficit disorder, hyperkinesia, aggressiveness, irritability, negativism). The less favorable neuropsychological outcome might be related to additional interacting factors such as personal antecedents, seizure onset and antiepileptic treatment. PMID- 9408593 TI - Sandhoff disease in the Turkish population. AB - Eighteen cases affected by Sandhoff disease were investigated by an enzymatic study of serum and leukocytes during the period 1988-1996, the clinical expression and enzymatic study were reported and discussed. An indirect minimum disease incidence was calculated in the Turkish population. Hexosaminidase activity in serum and leukocytes was severely deficient when measured by synthetic substrate 4-MU-N-acetylglucosaminide using the thermolabile fractionation procedure. Fractionation of hexosaminidase revealed different levels of isoenzymes A and B. Clinically, organomegaly was not found in 11 out of 18 infantile Sandhoff disease patients, while the remaining seven had mild organomegaly. Organomegaly was not found in patients with relatively high % hexosaminidase B activities. These results suggested that patients with different percent heat-stable enzyme activity may have a different type of mutation which is related to the underlying molecular heterogeneity in the Turkish population where 21% of marriages are found to be consanguineous. PMID- 9408595 TI - Early detection of axonal and neuronal lesions in prenatal-onset periventricular leukomalacia. AB - The expression of beta-amyloid precursor protein (beta-APP) immunoreactivity was investigated in 16 cases of prenatal-onset periventricular leukomalacia (PVL). beta-APP positive axons were found in the early stage of prenatal PVL, which included coagulation necrosis, microglial activation, axonal swelling or astrogliosis, but were not detectable in the late stage of prenatal PVL. Furthermore, beta-APP immunoreactive neurons were also observed in the fifth layer of pyramidal neurons of the cerebral cortex, corresponding to the beta-APP positive axons in PVL. Thus, beta-APP is detected as an early sign of axonal and neuronal lesions in prenatal-onset PVL, and neuronal beta-APP in the cerebral cortex may function to repair cell damage. In addition, prenatal PVL occurred at various stages before birth. PMID- 9408596 TI - Autonomic nervous system functions in children with nocturnal enuresis. AB - Nocturnal enuresis is involuntary urination during night sleep. The pathogenesis of nocturnal enuresis is controversial. Developmental delay, genetic factors, stress and psychological factors, and sleep abnormalities are considered to be the etiologic factors. Various urodynamic studies showed bladder hyperactivity in enuretic children. Since the cause of vesical hyperactivity is not clear, we investigated the possible role of autonomic nervous system dysfunction in these children. The study groups consisted of 41 enuretic (25 boys and 16 girls) and 30 healthy children (18 boys and 12 girls). Four non-invasive autonomic nervous system function tests (orthostatic test, Valsalva ratio, 30:15 ratio, heart rate responses to deep breathing) were carried out in both groups. The differences between the enuretic and control groups were statistically significant in the Valsalva and 30:15 ratios (P < 0.0005 and P < 0.005, respectively). The results of these two tests demonstrated parasympathetic nervous system hyperactivity. No abnormality of the sympathetic nervous system was found. We suggest that the parasympathetic nervous system hyperactivity shown in our study may be a cause of vesical hyperactivity in enuretic children. PMID- 9408597 TI - Early infantile Krabbe disease: deceptively normal magnetic resonance imaging and serial neurophysiological studies. AB - Early infantile Krabbe disease is a progressive neurodegenerative disease caused by deficiency of lysosomal enzyme galactocerebroside beta-galactosidase, with onset before the age of 6 months. We present serial clinical, radiological and neurophysiological findings of a patient with early infantile Krabbe disease, presenting at the third day of life with hypotonia, macrocephaly and neonatal seizures. The patient had a deceptively normal initial magnetic resonance imaging examination at the age of 3 months, with progression of the white matter disease over the following 9 months, showing a clinical picture of profound hypotonia with pyramidal and pseudobulbar signs, as well as mild optic atrophy. Assay of galactocerebroside beta-galactosidase activity in leukocyte culture disclosed a marked deficiency of the enzyme (0.00 nmol/mg protein per h with normal values > 0.7 nmol/mg protein per h), thus confirming the diagnosis of Krabbe disease. Nerve conduction velocity and evoked potential studies, as well as the electroencephalogram, were abnormal at the age of 6 months, while serial neurophysiological studies at the age of 12 and 18 months demonstrated the progressive nature of the disease. PMID- 9408598 TI - Classical Rett syndrome in sisters: variability of clinical expression. AB - Familial cases of Rett syndrome (RS) are rare. No significant differences have been reported in the clinical courses of concordant monozygotic twins with RS. We present the variability of clinical expression in two Japanese sisters with classic RS. The younger sister, currently 6 years and 6 months old, never stood or walked alone, showed severe spasticity, growth retardation, and microcephaly and developed sleep-wake rhythm disturbance from age 4 years and seizures from age 5 years. The elder, currently 7 years and 9 months old, walked alone and had mild spasticity, no growth retardation, normal sleep-wakefulness rhythm and no seizures. RS is most likely to be transmitted as an X-linked dominant, male lethal (XDML) disorder, although this is still contested. If RS is an XDML disorder, lyonization may account for variability of expression in the sisters. PMID- 9408599 TI - Antenatal thalamic lesions in the newborn: two anatomoclinical cases. AB - We report two cases of antenatal bilateral thalamic lesions constituted by neuronal loss, gliosis and mineralized glial or neuronal cells. No etiology could be found. Neuroradiological findings were poorly correlated with histological changes. These cases are compared with the few previously reported cases of the same condition. We strongly recommend extensive etiological investigation as recurrence occurred in one family. PMID- 9408600 TI - An autopsy case of bathtub drowning in epilepsy. AB - We report an autopsy case of bathtub drowning in epilepsy. A 26-year-old female with mental retardation had been treated for refractory epilepsy. Her younger sister found her floating supine in the bathtub 45 min after starting bathing. Neuropathological examination revealed cerebral cortical dysplasia in the precentral gyrus of the left hemisphere, which had not been detected by MRI, suggesting the etiology of epilepsy. In bathtub submersion injury of an unidentified cause, neuropathological examination should be performed to reveal any lesion underlying epileptic seizures. Additionally, we present statistics on bathtub submersion injury in children aged 5 years or older in Japan based upon nationwide survey data obtained in 1991. Forty-seven percent of them had associated epilepsy or convulsive attacks and 71% died. It is necessary for epileptic patients and their families to understand that the risk of bathtub drowning can be minimized if proper precautions are taken. PMID- 9408601 TI - Effective immunoglobulin therapy for brief tonic seizures in methylmalonic acidemia. AB - We report on a patient with methylmalonic acidemia (MMA). He experienced a metabolic acidosis attack at 3 weeks of age. He immediately received peritoneal dialysis and exchange transfusion, and recovered from the attack. His MMA phenotype was mut0. Dietary therapy (strict protein restriction) was found to be effective in preventing further attacks, and he had mild hypotonia and impaired psychomotor development. At 9 months of age, he developed brief tonic seizures, which showed polyspike bursts under EEG. His psychomotor development continued to deteriorate. However, intravenous administration of immunoglobulin (200 mg/kg/day for 5 consecutive days) had a dramatic effect; his seizures disappeared and his psychomotor development improved. PMID- 9408602 TI - Benzodiazepine (BDZ) treatment of benign childhood epilepsy with centrotemporal spikes (BECCT) PMID- 9408603 TI - Inhibition of establishment of hepatic metastasis in mice by combination gene therapy using both herpes simplex virus-thymidine kinase and granulocyte macrophage-colony stimulating factor genes in murine colon cancer. AB - Herpes simplex virus-thymidine kinase (HS-tk) gene therapy with ganciclovir (GCV) treatment has been reported to inhibit the tumor growth, which is applied to the gene therapy targeted to the malignant brain tumor. To suppress the tumor growth completely, the authors designed the HS-tk gene therapy in combination with granulocyte macrophage-colony stimulating factor (GMCSF) gene using the hepatic metastatic model of murine colon cancer. The transduction of the HS-tk gene in combination with the GMCSF gene, followed by GCV, showed a complete inhibition of hepatic metastases of murine colon cancer, which was significantly superior to that of HS-tk gene alone. The growth of cancer cells transduced with both HS-tk and GMCSF genes was inhibited in vitro, and long-lasting antitumor immunity after hepatic metastasis of cancer cells transduced with both HS-tk and GMCSF genes was acquired. It is suggested that HS-tk gene therapy in combination with the GMCSF gene is effective for the complete inhibition of hepatic metastasis of murine colon cancer. PMID- 9408604 TI - Vaccination for experimental gliomas using GM-CSF-transduced glioma cells. AB - Brain tumors have an immunoprivileged status which contributes to their refractoriness to treatment. In this study, immune rejection of GL261 glioma tumors in the mouse brain was achieved by subcutaneous vaccination with GM-CSF transduced glioma cells. Cultured GL261 cells were transduced to secrete murine GM-CSF using a retrovirus vector, then irradiated, and injected subcutaneously into H-2 matched C57BL/6 mice. In prevaccination studies, the median survival time (MST) of animals vaccinated with 5 x 10(4) or 5 x 10(5) GM-CSF-transduced cells 7 days prior to intracranial injection of 10(6) nontransduced, nonirradiated GL261 cells was significantly prolonged by 45-50% compared with animals vaccinated in parallel with nontransduced, irradiated glioma cells. In treatment of established gliomas, the MST of animals, which were treated subcutaneously with 5 X 10(6) irradiated GM-CSF-transduced cells 3 days after intracranial injection of 2 x 10(4) nontransduced cells, was prolonged significantly by 36% compared with animals treated with the same number of nontransduced, irradiated cells or to sham-treated animals. In prevaccination studies, histology of brain tumors 4 days after intracranial tumor cell injection revealed infiltrates of CD8+ lymphocytes and eosinophils, the latter exclusively in animals vaccinated with GM-CSF-transduced cells, Thus, subcutaneous injection of irradiated GM-CSF-transduced glioma cells can induce a potent immune response to intracranial gliomas both as a vaccination against subsequent intracranial glioma cell implantation and for treatment of established intracranial glioma. PMID- 9408605 TI - Inhibition of established tumor growth in syngeneic mice by local inoculation of engineered mouse myeloma cells secreting a recombinant fusion protein RM4/TNF. AB - Mouse myeloma cell line VKCK/RM4-TNF secreting the recombinant fusion protein RM4/TNF was used to study the relationship between tumor necrosis factor (TNF) secretion of tumor cells and its tumorigenicity, and to study the potential mechanisms responsible for the antitumor immune response. To evaluate tumorigenicity, 5 x 10(5) viable TNF-secreting VKCK/RM4-TNF and non-TNF-secreting VKCK tumor cells were subcutaneously injected into BALB/c mice. Tumor progression or regression was evaluated 2 weeks after tumor inoculation. Our animal studies showed that RM4/TNF secretion by VKCK/RM4-TNF tumor cells curtailed its tumorigenicity in BALB/c mice and induced a long-term, protective immune response against a subsequent challenge of the parental VKCK tumor cells. Our animal studies in T-cell subset-depleted BALB/c mice and in T-cell-deficient nude mice demonstrated that both CD4+ and CD8+ T cells play a major role in the reduction of tumorigenicity. In addition, our results further showed that local inoculation of irradiated VKCK/RM4-TNF cells secreting TNF was able to significantly inhibit the established VKCK tumors in syngeneic mice. This study thus highlights the potential utility of engineered tumor cells secreting RM4/TNF in cancer gene therapy. PMID- 9408607 TI - In vitro and in vivo characteristics of human squamous cell carcinoma of the head and neck cells engineered to secrete interleukin-2. AB - Two human squamous cell carcinoma of the head and neck (SCCHN) cell lines, PCI-13 and PCI-52, were transduced with the retroviral construct containing human interleukin-2 (IL-2) cDNA and selected for neomycin resistance in G418 medium. Stably transduced SCCHN cells produced and secreted IL-2, which was shown to have biologic activity in a bioassay, using an IL-2-dependent CTLL-2 cell line. By immunohistochemistry, IL-2 gene-transduced PCI-13 cells were strongly positive for IL-2, and by flow cytometry showed both cell surface and intracytoplasmic expression of IL-2 protein. Expression of IL-2 mRNA was measured by quantitative RT-PCR and found to be considerably increased in transduced SCCHN relative to that in parental cells. There was no difference in expression of IL-2R between the parental and IL-2 gene-transduced cells. In vitro proliferation of IL-2 gene transduced tumor cells was consistently more rapid than that of parental cells. Sensitivity of the parental and IL-2 gene-transduced targets to lysis or apoptosis mediated by purified human natural killer (NK) cells or IL-2-activated NK (A-NK) cells was comparable as measured in 4-hour 51Cr-release and 1-hour [3H]thymidine-release assays, respectively. However, transduced cells were significantly more sensitive than parental cells to these effectors in 24-hour MTT assays, most likely due to IL-2 production by the transduced targets. PCI-52 cells selected for in vivo experiments formed large subcutaneous tumors in immunosuppressed nude mice. Tumors established by subcutaneous injections of 1 x 10(7) IL-2 gene-transduced cells regressed completely by day 25, while those formed by parental or LacZ gene-transduced tumor cells grew progressively. Tumor regression was mediated by numerous mononuclear cells, identified as murine NK cells and macrophages by immunohistochemistry, which accumulated around the IL-2 secreting, but not parental, tumors within 5-6 days after tumor cell injections. Thus, IL-2 gene-transduced SCCHN cells produce functional IL-2 in vivo in amounts sufficient to support the recruitment to the tumor site and antitumor activity of cytotoxic effector cells. IL-2-secreting SCCHN cells may be a useful component of vaccines designed to induce and sustain effector cell activation at the tumor site. PMID- 9408606 TI - Chemogene therapy: osteocalcin promoter-based suicide gene therapy in combination with methotrexate in a murine osteosarcoma model. AB - We previously reported that the recombinant adenovirus (Ad) vector containing the thymidine kinase (TK) gene driven by the osteocalcin (OC) promoter (Ad-OC-TK), when delivered concurrently with acyclovir (ACV), is highly selective in blocking the growth of osteosarcoma in experimental models (Cancer Res. 1996;56:4614 4619). To investigate the possible additive effects of the combined treatment of gene therapy and conventional chemotherapy (chemogene therapy), we compared the effect of low dose (IC10) methotrexate (MTX) and OC promoter-based toxic gene therapy with either of these single modalities alone. We choose low dose MTX with the intent of determining whether chemosensitization of the osteosarcoma may be possible in combination with gene therapy with an overall reduced toxicity profile and enhanced therapeutic efficacy when compared to a single agent alone. In vitro, the combined treatments of MTX (3 ng/mL) and Ad-OC-TK (20 multiplicity of infection (MOI)/target cell) plus ACV (10 mg/mL) had an additive therapeutic effect over that of either MTX (P < 0.05) or Ad-OC-TK plus ACV treatment alone (P < 0.05). In vivo, nude mice with subcutaneous tumors of either human osteosarcoma (MG-63) or rat osteosarcoma (ROS) received three intratumoral injections of Ad-OC TK (5 x 10(8) PFU) plus daily intraperitoneal ACV (40 mg/kg body weight) for 2 weeks in combination with five weekly bolus intraperitoneal MTX (87.5 mg/kg). Osteosarcoma tumor growth was inhibited more efficiently than by either Ad-OC-TK plus ACV (P < 0.05) or MTX treatment (P < 0.005) alone. At day 45 in the ROS group, 100% of the animals survived when treated with chemogene therapy, whereas 80% survived with gene therapy and no animals survived in the MTX-treated or untreated controls. In summary, we developed a novel therapeutic strategy for the treatment of osteosarcoma employing both chemotherapy and gene therapy. Chemogene therapy could potentially achieve better antitumor effects with reduced toxicity than the conventional chemotherapy or gene therapy protocols alone. PMID- 9408608 TI - Cervical carcinoma cells transfected with the CD80 gene elicit a primary cytotoxic T lymphocyte response specific for HPV 16 E7 antigens. AB - Cervical carcinoma is strongly associated with human papillomavirus (HPV) type 16, and the transforming viral genes E6 and F7 are steadily expressed by the tumor cells. Therefore these viral oncogenes may be regarded as tumor-associated antigens. Our previous studies showed that cervical cancer cells after introduction of the CD80 gene activated allogeneic cytotoxic T lymphocytes (CTLs). In this study, we tested whether HPV 16+ cervical tumor cells (CaSki) expressing CD80 were able to activate CTLs recognizing HPV 16 E7 antigen. To this end, CD80+ CaSki cells (HLA-A*0201+) were used to stimulate peripheral blood T lymphocytes from HLA-A*0201+ healthy donors. We found that the activated T cells were able to lyse parental CaSki cells as well as Epstein-Barr virus-immortalized autologous B cells loaded with HLA-A*0201-restricted E7 peptides (amino acids 11 19, 82-90, 86-93). In contrast, no lysis was observed against target cells loaded with a control HIV-reverse transcriptase peptide (amino acid 476-484, HLA-A 0201 restricted). Our data, for the first time, provide evidence that CD80-expressing cervical cancer cells are able to activate tumor-specific CTLs using HPV 16 E7 as tumor-associated antigens. PMID- 9408609 TI - A recombinant adenovirus expressing wild type p53 induces apoptosis in drug resistant human breast cancer cells: a gene therapy approach for drug-resistant cancers. AB - The cytotoxicity of a recombinant adenovirus expressing the wild type tumor suppressor gene p53 (AdWTp53) was studied in two human breast cancer MCF-7 sublines selected for resistance to adriamycin (MCF-Adr) and mitoxantrone (MCF Mito). Although the levels of wild type p53 protein following infection with AdWTp53 are comparable in all cell lines, the two drug-resistant MCF-7 sublines were 300- and 18-fold more sensitive to killing by AdWTp53 compared with the drug sensitive parental MCF-7 cell lines. In each cell line, AdWTp53 infection led to cell cycle arrest, and reduction of Cdk2 and cyclin B1-Cdc2 activity. Nucleosomal DNA fragmentation analysis (as a function of apoptosis) following AdWTp53 infection revealed that, while the parental MCF-7 cells failed to undergo apoptosis, both drug-resistant cell lines showed distinct DNA laddering. In MCF Adr cells, a combination treatment of AdWTp53 and adriamycin was much more toxic than either of the reagents used individually. Finally, exposure of a mixed population of MCF-Adr and CD34+ cells to AdWTp53 selectively prevented MCF-Adr cell colony formation, while there was no inhibition of CFU-GM colony formation from CD34+ cells. These findings suggest that some drug-resistant human breast cancers may be effectively treated with adenovirus expressing wild type p53. PMID- 9408610 TI - Cisplatin-based interferon gamma gene therapy of murine ovarian carcinoma. AB - Cis-diamminedichloroplatinum(II) increased in vivo lipofection efficiency of tumor cells with a target gene, interferon gamma (IFN gamma), in a murine metastatic ovarian carcinoma model. The expression of IFN gamma gene in ascitic tumors reached a peak at day 11 after cisplatin treatment and lasted over 1 week. A local increase in IFN gamma gene expression increased a local expression of inducible nitric oxide synthase (iNOS) and the tumor necrosis factor a gene. This treatment suppressed tumor growth (or killed tumor cells) and increased the long term survival of animals (at least 80 days) without tumors, which was prevented by simultaneous treatment of iNOS inhibitor aminoguanidine. The results indicate that cisplatin-based IFN gamma gene therapy is an effective treatment protocol and nitric oxide may play a major part for tumor cell killing or growth. PMID- 9408611 TI - Coronary artery stenting without anticoagulation, aspirin, ultrasound guidance, or high balloon pressure: prospective study of 1,051 consecutive patients. AB - Between March 1994 and November 1995, 1,212 coronary stents were implanted in 1,051 consecutive patients at our institution with the following protocol: daily pre- and poststenting treatment with ticlopidine 500 mg without aspirin, implantation under angiographic guidance, without ultrasound, with semi-compliant balloons inflated at 10 bars. Stenting was indicated after failure of balloon angioplasty (bail-out, dissection, elastic recoil) in 27% of the patients and considered as elective (de novo, restenosis, chronic occlusion, saphenous vein grafts) in 73% of the cases. During the 30-day follow-up period, stent thrombosis occurred in 11 patients (1.0%) and vascular access-site complications in three patients (0.3%). Thirteen patients (1.1%) died, 10 from previous left ventricular failure, 3 (0.3%) from subacute thrombosis. Multivariate analysis revealed that the size of the last balloon used was associated with subacute stent thrombosis Thus, in nonselected patients, placement of coronary stents may be safely achieved without use of warfarin, post procedural heparin, high balloon pressure, or ultrasound guidance. Antiplatelet therapy with ticlopidine and angiographic guidance result in a stent thrombosis rate of 1% and a vascular complication rate of 0.3%. PMID- 9408612 TI - Stenting without ASA? PMID- 9408613 TI - Complications of cardiac angiography using low- or high-osmolality contrast agents in patients with left main coronary stenosis. AB - Recently published guidelines suggest that, in view of cost concerns, low-osmolal contrast should be selectively used in patients at increased risk of experiencing a contrast-related complication during cardiac angiography. The suggested criteria include the presence of left main coronary disease. However, the presence of left main disease is not usually known prior to angiography. Contrast related complications of cardiac angiography were therefore analyzed in a group of 111 clinically stable patients found to have left main coronary stenosis, to determine if use of low-osmolality contrast had any beneficial effect when compared to standard contrast. Data were gathered prospectively as part of a randomized controlled trial, and the subgroup of patients with left main disease was analyzed retrospectively. Complications were divided into minor, intermediate, and major categories. In the 58 patients who received high-osmolar contrast, there were 4 contrast-related minor reactions, 8 intermediate events requiring treatment, and 1 major adverse event. Among the 53 patients who received low-osmolar contrast, there were no minor reactions, 7 intermediate events requiring treatment, and no major adverse events. The only difference of borderline significance was in the incidence of minor reactions requiring no treatment (P = 0.05). Although small and therefore not definitive, this study suggests that 1) universal use of low-osmolar contrast agents would not be expected to eliminate the risk of contrast-related reactions to cardiac angiography; 2) the well-documented clinical differences between high- and low osmolar contrast primarily involve mild reactions; and 3) standard high-osmolar contrast is reasonably safe in clinically stable patients with left main coronary stenosis. The results therefore are consistent with the notion that selective use of low-osmolar contrast only in unstable patients is safe and appropriate. PMID- 9408614 TI - Dealing with the highs and lows of contrast agents in left main disease. PMID- 9408615 TI - Effect of gender on outcomes following renal artery stent placement for renovascular hypertension. AB - To study the effect of gender on outcome following renal artery stent placement for renovascular hypertension, we prospectIvely followed 66 patients (30 males, 36 females) who underwent Palmaz stent placement in 89 renal arteries. There was no difference in the incidence of procedure-related complications between males and females. At 6-mo follow-up, the decrease in systolic (35 +/- 30 mm Hg and 27 +/- 25 mm Hg) and diastolic (15 +/- 23 mm Hg and 14 +/- 14 mm Hg) blood pressures was similar in female and male patients, respectIvely. Late follow-up at 19 +/- 11 mo also showed no difference in blood pressure response. In 44 patients who underwent repeat angiography at a mean duration of 9.1 +/- 5.6 mo after stent deployment, the incidence of restenosis was 26% in females and 24% in males (P = 0.85). We conclude that gender has no effect on the incidence of complications, blood pressure response, or angiographic restenosis in patients undergoing renal artery stent placement. PMID- 9408616 TI - Gender differences in cardiovascular treatment: renal artery stenting. PMID- 9408618 TI - Promises, promises... PMID- 9408619 TI - A round peg for a round hole--progression to the ideal! PMID- 9408617 TI - Transcatheter closure of secundum atrial septal defects using the new self centering amplatzer septal occluder: initial human experience. AB - Transcatheter closure of secundum atrial septal defect (ASD) using clamshell or buttoned devices is accompanied by a high incidence of residual shunt. Recently, a new self-centering device, the Amplatzer septal occluder (ASO), has been evaluated in an animal model with very good results. Therefore, our purpose is to report on our initial clinical experience with this device. Thirty patients underwent an attempt at catheter closure of their ASDs at a median age of 6.1 yr (range, 2.9-62.4 yr) and median weight of 22 kg (range, 13-69 kg) using the ASO. The median ASD diameter measured by transesophageal echocardiography (TEE) was 12.5 mm (range, 5-21 mm), and the median ASD balloon stretched diameter was 14 mm (range, 7-19 mm). All patients had right atrial and ventricular volume overload with a mean +/- SD Qp/Qs of 2.3 +/- 0.6. A 7F catheter was used for delivery of the device in all patients. The device was placed correctly in all patients. There was immediate and complete closure (C) in 17/30 patients, 10 patients had trivial residual shunt (TS), and 3 had moderate residual shunt (MS). The median fluoroscopy time was 15 min (range, 8-35 min), and the median total procedure time was 92.5 min (range, 40-135 min). There was no episode of device embolization or any other complication. Follow-up was performed using transthoracic echocardiography (TTE) 1 day, 1 mo, 3 mo, and yearly thereafter. At 1 day, there was C of the ASD in 24/30 patients, 3 had TS, 1 had small shunt (SS), and 2 had MS. At a median follow-up interval of 6 mo, there have been no episodes of endocarditis, thromboembolism, or wire fracture. We conclude that the use of the new ASO is safe and effective in complete closure of secundum ASDs up to a diameter of 21 mm in the majority of patients. Further clinical trials are underway. PMID- 9408620 TI - Transcatheter management of neonates with pulmonary atresia and intact ventricular septum. AB - This report describes a 1 day-old infant with valvar pulmonary atresia with intact ventricular septum in whom we were successful in performing transcatheter guidewire perforation and balloon pulmonary valvuloplasty to establish right ventricle-to-pulmonary artery continuity and flow. Also described is implantation of a 4 mm coronary stent into ductus arteriosus in lieu of surgical aortopulmonary shunt to treat pulmonary oligemia and systemic arterial hypoxemia. Details of transcatheter guidewire perforation are presented and it is suggested that this method be used as an alternative to Laser/radio frequency wires, especially in the absence of approval of the latter wires by the regulatory agencies. Stenting of the ductus may be considered an alternative to surgical aortopulmonary shunt. Role of transcatheter technology in the management of selected patients with pulmonary atresia and intact ventricular septum is discussed. PMID- 9408621 TI - To perforate or not to perforate-that's the question...or is it? just ask Richard! PMID- 9408623 TI - Interventionalists: watch out for coronary arterial anomalies in tetralogy of Fallot. PMID- 9408622 TI - Single coronary artery complicating stent implantation for homograft stenosis in tetralogy of Fallot. AB - Successful stent implantation for conduit stenosis has been described; however, this procedure may be complicated by compression of adjacent structures during expansion. We report on a rare case of a single right coronary artery system complicating stent implantation for relief of homograft stenosis in tetralogy of Fallot. PMID- 9408624 TI - Percutaneous retrieval of broken silastic catheter from the left atrium in a critically Ill premature infant. AB - A critically ill premature neonate (birthweight 1,395 g, gestational age 30 wk) had a broken silastic catheter lodged in the left atrium. We successfully retrieved the foreign body by percutaneous approach using a helical basket catheter under echocardiographic control. Such a therapeutical option for a broken, lightly radiopaque catheter has not been previously described in very low birthweight, critically ill infants. PMID- 9408625 TI - Aortic dissection occurring during coronary angioplasty: angiographic and transesophageal echocardiographic findings. AB - A localized acute aortic dissection was produced in 2 patients, complicating coronary angioplasty. In both cases a coronary dissection provided the entry door, with subsequent retrograde progression of the dissection into the aortic root. After sealing the entry door, both patients could be managed conservatively using transesophageal echocardiography to accurately define the location of the intimal flap and to rule out dissection progression. PMID- 9408626 TI - Aortic dissection--exceedingly rare complication of coronary angioplasty. PMID- 9408627 TI - Unsuspected articulation side-strut extension during stenting in a tortuous right coronary artery. AB - This case report demonstrates the effect of articulation stent-strut extension during attempts to deploy a stent in a tortuous, calcified right coronary artery. The manipulation of the stent-sheath relationship and the effect of a tortuous, calcified artery produced serial dissections resulting in the implantation of three stents to seal the dissection. Caution should be used when employing rigid sheathed stent systems through tortuous vessels. PMID- 9408628 TI - Coronary artery aneurysm development after successful primary stent implantation. AB - This case report describes a patient who developed an aneurysmatic dilation of a coronary artery 6 months after successful primary stent implantatIon. The dilation occurred within the stented segment of the artery. To our knowledge, this is the first report of the development of an aneurysm, in the absence of angiographically visible dissection or other possible causative factors. PMID- 9408629 TI - Aneurysm of the mitral-aortic intervalvular fibrosa as a rare cause of angina pectoris: angiographic demonstration. AB - Aneurysms of the mitral-aortic interventricular fibrosa (MAIF) are exceptionally rare complications, commonly following aortic valve endocarditis. This report describes the angiographic findings of such an aneurysm, in a patient who developed an uncommon symptomatology of unstable angina pectoris, caused by the aneurysm's expansion against the coronary arteries. Surgical treatment is also discussed. PMID- 9408631 TI - Anomalous origin of left main coronary artery from the noncoronary sinus: an intravascular ultrasound observation. AB - We report on a rare case of anomalous origin of left coronary artery from the noncoronary sinus of Valsalva. Intraaortic intravascular ultrasound study identified the origin of the left coronary artery and facilitated subsequent selective coronary angiography of the artery. PMID- 9408630 TI - Saphenous vein covered stenting for right coronary artery lesion containing thrombus. AB - Vein covered stenting to close coronary pseudoaneurysm and perforation and in the setting of acute myocardial infarction have been described. This case report describes saphenous vein covered stenting to exclude a large thrombus in a right coronary artery lesion. Vein covered stenting may be considered as an option when dealing with a thrombus containing lesion. PMID- 9408632 TI - Fatal stent thrombosis following successful treatment or coronary artery rupture in an octogenarian. AB - Primary intracoronary stenting of a calcified left anterior descending coronary artery stenosis was complicated by within-stent coronary artery rupture and subsequent cardiac tamponade. Despite pericardiocentesis and sealing of the perforation by additional stent placement, subsequent stent thrombosis resulted in anterior myocardial infarction and fatal cardiogenic shock. PMID- 9408633 TI - Anomalous origins of the left coronary arteries from the right sinus of valsalva with an unusual course. AB - We report and attempt to classify a previously undescribed coronary artery anomaly. Our patient had all three coronary arteries arising from a common ostium in the right sinus of Valsalva, with an unusual distribution to the left coronary artery system: the anomalous left circumflex system taking an intraseptal (intramyocardial) course and the anomalous left anterior descending system taking an interarterial (between the great vessels) course. PMID- 9408634 TI - Stent placement for recurrent vasospastic angina resistant to medical treatment. AB - The successful stent placement for treatment of recurrent vasospastic angina in a patient with nonstenotic coronary arteries is described. Use of the Palmaz-Schatz stent resulted in successful vasodilation that completely prevented anginal attacks. This procedure represents an alternative treatment for patients with vasospastic angina refractory to aggressive medical therapy. PMID- 9408635 TI - The fight against coronary spasm--one more weapon. PMID- 9408636 TI - Focal angioplasty: theory and clinical application. AB - Percutaneous transluminal coronary angioplasty (PTCA) has been performed with 20 mm long uni-diameter balloons for over 17 years with excellent results. It has now become possible to combine both compliant and non-compliant balloon material on a single device, allowing the interventionalist to direct the force of dilation to the lesion focally. This concept has been termed "focal angioplasty." This article reviews the concept of focal angioplasty, including the physics of balloon dilation, and describes current clinical applications for focal angioplasty. We discuss use of the focal angioplasty technique for PTCA and stenting and review the results of several ongoing clinical trials. While the acute results using focal angioplasty have been promising, long-term benefits have not been fully studied and will require completion of ongoing trials. PMID- 9408637 TI - New stent delivery balloon: a technical note. AB - This study reports the first clinical application of a new noncompliant balloon composed of a middle polyurethane layer sandwiched between an inner layer of polyethylene terephtalate and an outer membrane that provides for consistent even expansion. With this balloon design, the very low compliance and high pressure resistance of polyethylene terephthalate are associated with the high elasticity of polyurethane, preventing balloon damage from stent crimping and expansion and allowing a firm embedding of the stent struts. Palmaz-Schatz stent implantation was successful in 33/35 stents (94%), and the two stents that could not be advanced up to the lesion were successfully withdrawn. High pressure expansion of the stent was obtained during deployment with no balloon ruptures at inflation pressures equal or lower than 16 atmospheres (atm). Accurate positioning of the stent was facilitated by the two markers at the balloon ends and by the optimal visualization after contrast injection, even with 6 Fr guiding catheters. This new delivery system maintains the advantages of hand-crimped stents on noncompliant balloons, reducing the risk of stent loss. PMID- 9408638 TI - Coronary stent and over-the-wire catheter exchange using standard length guidewires: Jet exchange (JEX) practice and theory. AB - The practice and theoretical principles of hydraulic exchange of over-the-wire (OTW) stent and PTCA catheters are described. Seventy-eight Palmaz-Schatz coronary stent delivery systems (PS-SDS), 8 Cook Flex-stents, and 247 assorted OTW catheters were delivered and extracted over standard length coronary guidewires using Jet Exchange (JEX). JEX was performed by pressurizing the wire lumen of coronary stent catheters to 18-20 atm and PTCA catheters to 15 atm. Extraction and insertion times were measured in the last 10 PS-SDS and PTCA procedures. Mechanical analysis of JEX was performed for PS-SDS and a representative OTW PTCA catheter by solving the Navier Stokes equation for annular flow with changing geometry. The force/mass relationship, extraction time, average velocity, net force on the guidewire, and drag force on the guidewire were determined for varying pressures, catheter masses, and extraction wire lumen fluids. JEX was successful in 75/78 (96%) of coronary stents and in 243/247 (98%) of the PTCA catheter exchanges. After catheter removal, reinsertion of another OTW catheter was successful in 324/325 (99%) attempts. The mean force on the guidewire at 15 and 20 atm ranged from 16,000 to 22,000 dynes. Extraction velocity was approximately equal to 250% greater when saline was used compared to the more viscous 50/50 contrast-saline solution. Timed JEX extractions for the PS SDS and standard PTCA catheters were 8.9 +/- 2.3 sec and 5.3 +/- 1.4 sec and compared favorably to theoretical calculations of extraction times, 9.8 and 3.8 sec respectively. JEX is a simple, reliable, and cost effective means of rapidly exchanging OTW stent delivery and PTCA catheters without using exchange wires, extension, or wire trapping devices. Analysis of the principles of conservation of momentum provides a basis for understanding the physical laws that permit safe and expedient JEX in a clinically setting. PMID- 9408639 TI - Radial approach: what about the learning curve? PMID- 9408641 TI - Detection of pulmonary emboli and associated right heart dysfunction by combining ventilation perfusion lung scanning and xenon ventriculography. AB - PURPOSE: The authors sought to determine if the right ventricular ejection fraction (RVEF), as measured by xenon ventriculography, is depressed in patients with pulmonary emboli. The authors also sought to correlate any decrement in RVEF with the extent of lung perfusion defects. MATERIALS AND METHODS: The authors identified all patients who had lung ventilation-perfusion (V/Q) scans between January 1994 and December 1996, that were interpreted as high probability for pulmonary embolism. From these patients, the authors selected those who had undergone concurrent xenon ventriculography (XV) (n = 23), and then reprocessed the initial ventriculography data for confirmation. The authors also reviewed original V/Q scans, chest radiographs, and clinical data. A control group was drawn from patients with normal V/Q scans who had undergone XV. RESULTS: Fifteen patients (65%) with high probability V/Q scans had an abnormally low RVEF (< .32). Patients with high probability V/Q scans also had a significantly lower mean RVEF (0.28 +/- .08) than patients with normal V/Q scans (.39 +/- .08 SD). The degree of RVEF decline correlated poorly with the number of segmental perfusion defects (r = -.39). CONCLUSIONS: RVEF is often depressed in patients with high probability V/Q scans. XV can identify these patients, while routine lung V/Q scans cannot. PMID- 9408640 TI - In-111 octreotide imaging in staging of small cell lung cancer. AB - BACKGROUND: Small cell lung cancer (SCLC) tumors have neuroendocrine features. In vitro and in vivo studies have demonstrated that 50%-75% of SCLC tumors express receptors for somatostatin. This might enable in vivo localization of the primary tumor and its metastases by using scintigraphy with a radiolabeled somatostatin analogue, such as octreotide. PURPOSE AND METHODS: The efficacy of scanning with In-111 labeled octreotide (octreotide scan) was studied in the staging of SCLC patients and compared with the results of conventional staging (liver ECHO, bone scintigraphy, MRI of the brain, spine, and pelvis). Imaging was performed in 29 patients with histologically confirmed SCLC at 4, 24, and 48 hours after intravenous injection of 185 MBq In-111 octreotide. RESULTS: In 24 of 29 patients, the primary tumor was visualized. In these 24 patients, 26 metastases were demonstrated with conventional staging, of which only nine were visualized with octreotide scan. Octreotide scans showed two metastases in the brain that were not visualized by MRI. In the other five patients, five metastases were demonstrated with conventional staging. Only two of these were detected with octreotide scan. However, octreotide scan did show a further metastasis in the brain that was not visualized by MR imaging. CONCLUSION: Octreotide imaging has a limited use in the detection of SCLC metastases compared to conventional staging. It might have some specific value in the detection of brain involvement in patients with limited disease. PMID- 9408642 TI - Spinal bone SPECT in chronic symptomatic ankylosing spondylitis. AB - Bone SPECT has acquired a useful role in the evaluation of acute and chronic back pain. Spinal pain is also a characteristic of chronic ankylosing spondylitis. The authors investigated the bone SPECT appearances of the lower thoracic and lumbar spine in 28 symptomatic patients with established ankylosing spondylitis, half of whom had complete ankylosis of the sacroiliac and one third complete ankylosis of the intervertebral joints. SPECT abnormalities were identified in 89%. Facetal joint uptake was found in 16 (57%) patients, of whom 7 (25%) had three or more, and 2 (7%) had seven or more active sites. The only other common site of uptake was in the vertebral body, in which 15 patients (54%) showed increased uptake with three or more sites found in 3 (11%) patients. The authors conclude that SPECT abnormalities in the lower thoracic and lumbar spine are frequently found in patients with ankylosing spondylitis, with the most common sites of abnormality in the facetal joints or vertebral body. Multiple sites of uptake in the facetal joints maybe striking in such patients and has not been previously described. PMID- 9408643 TI - Characterization of hepatic adenoma with atypical appearance on CT and MRI by radionuclide imaging. AB - The appearance of hepatic adenomas on CT and MRI are highly variable because of their varied histopathology, and images of adenomas are at times indistinguishable from those of other hepatic tumors. The authors present two patients with hepatic adenomas with extremely atypical CT and MRI manifestations demonstrating a "nodule-in-nodule" appearance. Radionuclide imaging in both patients showed decreased Ga-67 uptake in the adenomas compared to normal liver, negative colloid (Tc-99m phytate) uptake in the adenomas, and early uptake and subsequent retention of Tc-99m PMT. This correctly characterized the unique histopathologic features of the resected tumors. Radionuclide imaging using a combination of radiotracers may play an important role in aiding the diagnosis of this rare benign tumor, despite variable CT and MRI appearances. PMID- 9408644 TI - Tc-99m tetrofosmin in breast carcinoma and axillary lymph node metastases: a comparative study with Tc-99m MIBI. AB - The potential of Tc-99m tetrofosmin for the imaging of breast carcinoma and axillary lymph node metastases was investigated and compared with that of Tc-99m MIBI. Thirty female patients with palpable breast masses underwent Tc-99m MIBI scintigraphy; 17 of those underwent Tc-99m tetrofosmin scintigraphy. The axillary and breast regions were evaluated in all patients. All patients underwent biopsy within 2 weeks of the study. Twenty patients were found to have a primary malignancy of the breast, whereas 10 had benign disease. The patients with breast carcinoma had surgery. Twelve patients had axillary lymph node metastases. Tc-99m MIBI breast imaging showed abnormal uptake in 18 of 20 malignancies and in 8 of 12 axillary lymph node metastases. Tc-99m tetrofosmin breast imaging showed abnormal uptake in 13 of 14 malignancies and in 6 of 10 axillary lymph node metastases. Sensitivity, specificity, and accuracy values obtained with Tc-99m MIBI and Tc-99m tetrofosmin scintigraphy for breast carcinoma were 90%, 90%, 90%, and 93%, 100%, 94%, respectively. The values obtained with Tc-99m MIBI and Tc-99m tetrofosmin scintigraphy for axillary lymph node metastases were 66%, 100%, 86%, and 60%, 100%, 76%, respectively. The authors conclude that both of these techniques are effective in the differentiation of malignant breast masses from benign ones and in detecting axillary lymph node metastases. However, Tc-99m tetrofosmin is superior to Tc-99m MIBI in detecting breast carcinoma. PMID- 9408645 TI - Tl-201 positive, Ga-67 negative hepatoblastoma: a case report of a 12-year-old boy. AB - Hepatoblastoma is a primary liver neoplasm in which prompt diagnosis and resection are critical to long-term survival. Nuclear scintigraphy plays an important role in the characterization of hepatic masses. The authors present an unusual case of hepatoblastoma in a 12-year-old boy in whom Ga-67 scintigraphy and serum alpha-fetoprotein were negative. Positive Tl-201 scintigraphy pointed toward the true malignant nature of the mass and should be considered in the investigation of hepatic masses in childhood. PMID- 9408646 TI - Renal SPECT with Tc-99m DMSA in children with upper urinary tract infections using a triple-headed gamma camera. AB - Renal cortical scintigraphy with Tc-99m DMSA provides an excellent imaging modality for the assessment of cortical damage secondary to upper urinary tract infection (UTI). METHODS: The authors evaluated 48 children with UTI, 12 of whom had a history of vescico-ureteral reflux (from first of fourth degree), by planar scintigraphy and SPECT using a triple-headed gamma camera equipped with parallel hole, high-resolution collimators. RESULTS: SPECT images yielded positive findings in 36 kidneys, whereas planar scans yielded positive findings in 18 kidneys. The total number of lesions detected by SPECT was 51, whereas the number found with the planar technique was 23. CONCLUSION: This study demonstrates the superiority of SPECT scanning in detecting kidney lesions in children with UTI. PMID- 9408647 TI - Fascial Tc-99m MDP uptake in eosinophilic fasciitis as demonstrated by SPECT. AB - BACKGROUND: The presence of bone-seeking radiopharmaceutical uptake in extraskeletal tissues of the lower or upper extremities may indirectly reflect the presence of active inflammatory lesions in patients in whom systemic disease is suspected. MATERIALS AND METHODS: The authors present the case of a 26-year old woman who had mixed signs of scleroderma and cosinophilic fasciitis, in whom misleading findings on planar bone scintigraphy suggested diffuse muscular tracer uptake in the lower extremities. RESULTS: However, using additional SPECT imaging of the pelvis and thighs, it was shown that the soft tissue radioactivity was clearly restricted to the fascia overlying the muscles. The fascial localization of the inflammation was confirmed by biopsy. CONCLUSION: SPECT imaging was proven useful in indicating the exact localization of an active inflammatory process in the muscle fascia. PMID- 9408648 TI - Neurofibromatosis type 2 (bilateral acoustic schwannomas) demonstrated by Tc-99m (V) DMSA SPECT. AB - Tc-99m (V) DMSA clearly demonstrated several cranial meningiomas, bilateral acoustic neurinomas and multiple subcutaneous neurofibromas in a patient of neurofibromatosis type 2 (NF-2). The present paper describes accumulation of Tc 99m (V) DMSA in cranial schwannomas and meningiomas as well as multiple peripheral neurofibromas. PMID- 9408649 TI - Tc-99m ECD SPECT imaging in aphasia caused by subcortical infarct. PMID- 9408650 TI - Tc-99m labeled white blood cell imaging in chronic pancreatitis with pseudocyst. PMID- 9408651 TI - Psoriatic dactylitis: bone scintigraphic appearances. PMID- 9408652 TI - Artifact of I-131 whole-body scan with thoracic vertebral uptake in a patient with papillary thyroid carcinoma. PMID- 9408653 TI - Tc-99m sestamibi scintigraphy of double parathyroid adenoma. PMID- 9408654 TI - Specific muscle uptake on bone scintigraphy after strenuous activity. PMID- 9408655 TI - Osteopetrosis (Albers-Schonberg): appearance on three-phase bone scintigraphy. PMID- 9408656 TI - Symmetric tibial stress fractures from extensive jump roping. PMID- 9408657 TI - Is the cortical blindness due to diaschisis? PMID- 9408658 TI - Spinal metastases of a high grade astrocytoma visualized with FDG-PET. PMID- 9408659 TI - Urethro-scrotal fistula presenting as massive scrotal "bladder" on radionuclide bone scan. PMID- 9408660 TI - Tc-99m labeled WBC imaging of lower extremity abscesses and skin necrosis due to skin popping. PMID- 9408661 TI - Retroperitoneal hemorrhage detected by RBC scintigraphy. PMID- 9408662 TI - Tc-99m sestamibi and gated blood pool scintigraphy in localization of ectopic mediastinal parathyroid. PMID- 9408663 TI - Ga-67 uptake in a perisplenic fluid collection: planar and SPECT images. PMID- 9408664 TI - Successful Tc-99m MIBI imaging of recurrent hyperparathyroidism in parathyroid autotransplanted patients. PMID- 9408665 TI - The eight world congress of the International Rehabilitation Medicine Association. PMID- 9408666 TI - Practical guidelines for independent assessment in randomized controlled trials (RCTs) of rehabilitation. AB - OBJECTIVE: To provide guidelines for conducting independent assessments in randomized controlled trials (RCTs) of rehabilitation. The article aims to assist those who plan to put independent assessment into practice and to be an introduction for those who are new to independent assessment. ISSUES: Possible causes of unblinding to group allocation are discussed, such as unblinding by other people, the environment, patients and assessors themselves. Other issues discussed in this paper are bias during assessment and monitoring levels of unblinding. CONCLUSION: The importance of monitoring levels of unblinding is stressed. Although it may not be possible to always keep an assessor blind in RCTs of rehabilitation, we should strive for perfection. It is therefore advocated that levels of unblinding should be reported. PMID- 9408667 TI - Early supported hospital discharge following acute stroke: pilot study results. AB - OBJECTIVE: To establish the feasibility and method of evaluation of an early supported hospital discharge policy for patients with acute stroke. DESIGN: A randomized controlled trial comparing an early supported discharge service to conventional care. SETTING: Three acute hospitals in Newcastle upon Tyne. SUBJECTS: Ninety-two eligible patients with acute stroke admitted between 1 February 1995 and 31 January 1996. MAIN OUTCOME MEASURES: Placement, length of stay, readmission rates, mortality, functional ability (Nottingham Extended Activities of Daily Living (ADL) Scale), handicap (Oxford Handicap Scale), global health status (Dartmouth Coop Function Charts) and carer stress (General Health Questionnaire 30 item). RESULTS: The median length of stay for patients randomized to early supported discharge was 13 days compared to 22 days in the conventional care group (p = 0.02). The median Barthel ADL index at seven days post stroke of patients randomized to early supported discharge was 15, and 13 for those randomized to conventional care (NS). At three months post stroke the median Nottingham EADL score of patients randomized to early supported discharge was 10 compared to 7 for those who received conventional care (NS). There were no statistically significant differences in the global health status of patients or carer stress. CONCLUSION: An early supported discharge service following acute stroke with individualized rehabilitation in the community is feasible and can be evaluated by a randomized controlled trial but a larger multicentre trial is needed before such a service is widely adopted. PMID- 9408668 TI - The effect of botulinum toxin on hand function after incomplete spinal cord injury at the level of C5/6: a case report. AB - OBJECTIVE: To investigate the benefits of the focal use of botulinum toxin in spasticity in the forearm seen after incomplete spinal cord injury. DESIGN: A single case study with standardized assessment before and at three-week intervals after injection. INTERVENTION: EMG-guided selective injection of botulinum toxin. SUBJECT: A 23-year-old man, 18 months post injury. MEASURES: Rivermead Motor Assessment; grip strength; Jebsen hand tests; visual analogue scale; Ashworth spasticity scale. RESULTS: Weakness was seen as expected with some functional losses, but the patient made gains in the areas of concern: shaking hands, typing, using the hand to drink. These gains were sustained at 12 weeks. CONCLUSION: Selective use of botulinum toxin to weaken muscles can lead to functional benefit. PMID- 9408669 TI - A qualitative study of specialist nurse support for stroke patients and care givers at home. AB - BACKGROUND: The involvement of five specialist nurses in providing a stroke support service was evaluated quantitatively in a recent randomized controlled trial. This complementary study used qualitative methods to evaluate trial outcomes more comprehensively. AIMS: To identify whether the nurses' intervention may have influenced the process of stroke recovery. METHOD: A purposefully selected subsample of 30 patients and 15 care-givers were interviewed within 1-3 months of their final quantitative assessment (12 months after recruitment to the randomized trial). Fifteen of the patients and eight of the care-givers had received visits from a specialist nurse. A semistructured interview was designed to include questions on perceptions of the recovery process and evaluation of services received. RESULTS: Some differences were evident between the accounts of control and intervention group subjects. The less tangible aspects of nurses' interventions--concern, attention, empathy and interest, when combined with sound professional knowledge, had identifiable value to the patients and care-givers. It appeared that the nurses had employed considerable sensitivity and skill in identifying and responding to particular needs at appropriate times. CONCLUSION: The qualitative evaluation offers a different picture to the quantitative results of the randomized controlled trial. In general, the findings of the qualitative study are more positive and encouraging than the quantitative results. The majority of patients and care-givers in the intervention group believed that they had benefited from the specialist nurse's visits. PMID- 9408670 TI - A pilot study to determine the efficiency of lightweight carbon fibre orthoses in the management of patients suffering from post-poliomyelitis syndrome. AB - OBJECTIVES: To assess the usage of carbon fibre orthoses in assisting post-polio syndrome patients who reported advancing weakness and difficulty in maintaining their independent ambulatory status. DESIGN AND INTERVENTIONS: Carbon fibre orthoses that are 30% lighter than metal braces were constructed. SUBJECTS: Thirty patients who complained of difficulty in maintaining their ambulation using long-leg braces were provided with lightweight orthoses. RESULTS: Seventy per cent of the patients reported satisfaction with their braces. There were, however, problems in construction and use. CONCLUSIONS: Ambulation can be preserved in some patients who report advancing weakness because of post-polio syndrome. PMID- 9408671 TI - The use of standardized outcome measures in rehabilitation centres in the UK. AB - OBJECTIVE: To ascertain which standardized instruments are currently most commonly used as outcome measures for rehabilitation in routine clinical practice in the UK. DESIGN: The study used a postal questionnaire which was sent out to members of two major societies of rehabilitation professionals in the UK. RESULTS: Of 182 rehabilitation centres represented by respondents, 140 (77%) collected at least one standardized measure and 42 did not. Principal reasons for not recording measures were lack of time and not knowing what to collect. As had been anticipated, a very wide range of different measures were used by different centres, however some clear favourites emerged including the 10 m Walk, the Motricity Index and the Nine-hole Peg Test. One hundred and twenty-three centres used one or more global disability measure of which the commonest were the Barthel index or one of its modifications and the functional Independence Measure (FIM) and/or Functional Assessment Method (FAM). Among units that used handicap or extended activities of daily living (EADL) scales, the Nottingham EADL, the London Handicap Scale and the General Health Questionnaire (GHQ-22 or 12) were most popular. Outside neurorehabilitation, the Harold Wood/Stanmore mobility grades were used by 10/18 amputee rehabilitation centres and the Health Assessment Questionnaire (HAQ) was used by 15/48 units providing musculoskeletal rehabilitation. CONCLUSIONS: It is clear that no one measure is suitable in all settings and services, but the most popular measures from this survey may reasonably form the basis for a 'basket of recommended instruments' that may help to guide units wishing to collect outcome data but not knowing which to choose. PMID- 9408672 TI - Joint angle measurement: a comparative study of the reliability of goniometry and wire tracing for the hand. AB - OBJECTIVES: To compare the inter- and intra-rater reliability of goniometry and wire tracing in the assessment of finger joint angles: metacarpo-phalangeal (MCPJ), proximal (PIPJ) and distal interphalangeal joints (DIPJ). DESIGN: Twenty occupational therapists and 20 physiotherapists with a range of clinical experience were recruited from nine different centres. Using a masked goniometer and wire tracing they carried out repeated assessments of the MCPJ, PIPJ and DIPJ of a normal subject fixed in two different positions. RESULTS: The two assessment methods did not produce comparable angle measurements. Goniometry showed greater inter- and intra-rater reliability than wire tracing. Regardless of the assessment tool, the repeatability coefficient indicated that DIPJ measurement was less reliable than the other joints. Clinical and specialist experience did not affect reliability. CONCLUSION: Although both goniometry and wire tracing show limitations as reliable assessment tools, it is recommended that where possible goniometry should be used. PMID- 9408673 TI - Assessing and restoring function in elderly people--more than rehabilitation. AB - OBJECTIVE: To quantify the medical component of assessing and restoring function in the elderly and to determine if a subset who did not require medical input could be identified. DESIGN: Prospective consecutive sample. SETTING: Inpatient assessment, treatment and rehabilitation unit for the elderly in New Zealand. PATIENTS: Two hundred patients aged 60-98 years. MAIN OUTCOME MEASURES: (1) Proportion of elderly people receiving rehabilitation who also required treatment of medical problems. (2) Characteristics of those not requiring medical input. RESULTS: One hundred and seventy-two (86%) of people required some medical input during their hospital stay for rehabilitation. Forty-nine per cent had medical conditions which had a direct impact on their rehabilitation. Rehabilitation was delayed for a mean 16.8 (13.0-20.6)% of hospital stay when significant medical problems required treatment. People with previously undiagnosed conditions affecting rehabilitation tended to be older. The referral diagnoses most likely to result in an uncomplicated inpatient stay (i.e. rehabilitation only or rehabilitation with treatment of minor medical problems) were postsurgical treatment (55% uncomplicated), stroke (47% uncomplicated) or amputation (55% uncomplicated). However, 45-53% of people with these problems had a major medical component to their stay. People with other referral diagnoses or those admitted from institutions for the elderly were more likely to have a medical component to their stay. CONCLUSION: Restoration of function in the elderly requires a combination of both medical and rehabilitation skills. Reliable predictors of those not requiring medical input could not be found. PMID- 9408674 TI - A study of the attitudes of a rural Indian community toward people with physical disabilities. AB - OBJECTIVE: To examine the attitudes of a rural community in southern India toward its disabled members. DESIGN: Prospective cross-sectional study. METHOD: One hundred and twenty randomly selected villagers around the town of Vellore in southern India were invited to take part in the study. The attitudes towards individuals with physical disabilities were examined using a modified version of the Scale of Attitudes Toward Disabled Persons (SADP). The questionnaire was administered in its original English format with simultaneous translation into Tamil. RESULTS: Data were complete in 111 cases, 82% of whom showed a positive attitude toward people with disability. Gender and employment status did not appear to have an influence on whether the respondent regarded disabled people in a positive or negative way but most older individuals expressed prejudice and/or challenged the rights of disabled people to equal opportunities in education, employment and social integration. Further data analysis demonstrated that more subjects with than without a disabled family member supported the rights of disabled individuals to education, employment and social integration. By contrast, the items of SADP which described the personal attributes of disabled people did not discriminate between the two groups. CONCLUSION: The attitudes of the rural community studied toward people with disabilities were unrelated to the sex or educational status of the respondents but was influenced by their age. This suggests that older subjects in rural communities in developing countries should be the main target for educational programmes which promote positive images of disability. PMID- 9408675 TI - The Barthel Index and its relationship to nursing dependency in rehabilitation. AB - OBJECTIVES: To investigate the clinical application of the Barthel Index as an indicator of nursing dependency in a younger disabled unit. DESIGN: A prospective study of 132 patients (mean age 49) with chronic neurological problems. SETTING: A younger disabled unit at a District General Hospital, Oxford. MAIN OUTCOME MEASURES: The Barthel Index, and total nursing hours. RESULTS: The Median Barthel Index was 7 (95% confidence interval 6-9). The mean nursing hours were 2.7 +/- 1.7 (CI 2.41-2.99). Spearman rank order correlation coefficient between the Barthel Index and nursing hours was r = -0.69 (CI -0.79 to -0.59). CONCLUSIONS: Regression analysis showed that it was possible to use the Barthel Index as an indicator of nursing dependency particularly in physical care, and that it was independent of age and diagnosis. PMID- 9408676 TI - Further validation of the Elderly Mobility Scale for measurement of mobility of hospitalized elderly people. AB - OBJECTIVE: To further assess the validity and inter-rater reliability of the Elderly Mobility Scale (EMS). Also whether the scale reflects elderly people's perceptions regarding their mobility, and whether it can predict discharge destination, or likelihood of falling. DESIGN: Questionnaire-based study completed on admission and weekly after this on all patients referred to physiotherapy for mobility problems over the course of one month. SETTING: Care of the elderly wards in the Bristol General Hospital. SUBJECTS: Sixty-six patients (ages 66-69 years, 66% female) were included in the validity study. Nineteen patients (ages 71-95 years, 47% female) were included in the inter-rater reliability study. INTERVENTIONS: EMS, Barthel and patients' perceptions of mobility were tested with routine physiotherapy treatment carried out as necessary. MAIN OUTCOME MEASURES: Concurrent validity was assessed by correlating EMS scores with Barthel scores using Spearman's test. Inter-rater reliability was also tested using a Spearman's correlation. EMS scores of patients were also evaluated in conjunction with whether or not they fell and their destination on discharge. RESULTS: A significant correlation between EMS and Barthel scores indicated concurrent validity. Inter-rater reliability was demonstrated on 19 patients with a significant correlation between scores. No predictive validity could be ascribed to EMS in terms of discharge destination or likelihood of falling. Results do indicate a possible predictive validity of the functional reach component of the EMS regarding the risk of future falls. CONCLUSIONS: The EMS was found to be a valid scale with good inter-rater reliability that could be readily applied during daily clinical work. However, it was found to have no predictive validity in terms of falling or discharge destination. PMID- 9408677 TI - Emergency day hospital assessments. AB - OBJECTIVE: To describe the use of a geriatric day hospital for emergency multidisciplinary assessment as an alternative to immediate hospital admission for some groups of frail older people. DESIGN: Retrospective study of change in clinical practice. SETTING: District general hospital. INTERVENTION: Immediate multidisciplinary assessment of selected patients instead of requested emergency hospital admission. OUTCOMES: (1) Data were collected on 67 consecutive referrals. In 63% the patient needs could be met without emergency hospital admission on the day of assessment. (2) Of the 42 patients originally not admitted only 29% were subsequently admitted within the next three months. CONCLUSIONS: Emergency day hospital assessments can be used as an alternative to immediate hospital admission for some frail older people. PMID- 9408678 TI - Bed rails: a barrier to independence? PMID- 9408679 TI - Subjective swallowing difficulties following stroke: a questionnaire survey. PMID- 9408680 TI - Multicultural issues in organ transplantation: the influence of patients' cultural perspectives on compliance with treatment. AB - The potential influence of racially or ethnically diverse cultural perspectives on patient compliance with post-transplant treatments is discussed. Domains of competency regarding culturally sensitive clinical practice are outlined to assist providers in better understanding the perspectives that may influence the views and behavior of culturally diverse patients. PMID- 9408681 TI - Commercially motivated renal transplantation: results in 540 patients transplanted in India. The Living Non-Related Renal Transplant Study Group. AB - Commercial renal transplantation is widely condemned on moral and ethical grounds. Moreover, previous studies involving small numbers of patients, mostly from tertiary care centers, have reported poor patient and graft survival. The Living Non-Related Renal Transplant Study Group was established to determine the overall results and complications in a large number of patients who received this type of transplantation in India. In this study we retrospectively collected data from 22 centers on 540 patients who had received commercial renal transplantation (CRT) in India between 1978 and 1993. The data collected included demographic characteristics of patients, the primary renal disease, dialysis and pretransplant data, immunosuppressive regimen, rejection episodes, post transplant data, as well as patient and graft outcomes. Results were compared with those of 75 recipients of emotionally related renal transplantation (ERT) performed in two participating institutions in the Middle East. Among the 540 patients, 7.5% were younger than 20 yr of age and 14.2% were 50 yr of age or older. After adjusting for several variables, the 1-, 3-, and 5-yr patient survival rates in the CRT were 97, 94, and 92%, and in the ERT 95, 91, and 91% respectively (p = 0.4921). The corresponding rates for graft survival were 90, 81, and 72%, and 90, 86, and 83%, respectively (p = 0.5336). There was a higher incidence of human immune deficiency virus (HIV) infection (4.6% vs. 0% p = 0.053), and hepatitis B virus (HbsAg) infection rate (8.1% vs. 1.4% p = 0.03) in CRT. In conclusion, patient and graft survival rates in recipients of commercial renal transplantation are similar to those in recipients of emotionally related transplantation done in our institutions. However, the incidence of serious infections with HIV and HbsAg is significantly higher. PMID- 9408682 TI - Lipoprotein pattern in end-stage renal disease and following successful renal transplantation. AB - Lipoprotein abnormalities are common in uremia and frequently persist after successful renal transplantation (RT). Based on the influence of immunosuppression on lipoprotein metabolism, this prospective study in 76 patients has been focused on lipoprotein pattern in end-stage renal disease and after successful RT during a follow-up period of 18 months. Additionally, the influence of different immunosuppressive regimes has been evaluated. Hyperlipidemia was present in 32% of the patients before and in 59% after grafting. Total cholesterol was 5.42 +/- 1.57 mmol/l (mean S D) before RT and continuously and significantly increased during the entire observation period, being highest 18 months after RT (6.8 +/- 1.63 mmol/l; p < 0.01). This was mainly due to an increase in LDL-cholesterol (before RT: 3.68 (1.41 mmol/l; 18 months after RT: 4.69 +/- 1.88 mmol/l; p < 0.05) while HDL-cholesterol values only slightly increased (before RT: 0.99 +/- 0.36 mmol/l; 18 months after RT: 1.13 +/- 0.3 mmol/l; p < 0.05). Changes in total HDL-cholesterol were mainly due to an increase in HDL 3, while HDL 2 moderately increasing remained at low concentrations. As corticosteroid dosage was substantially reduced during the entire observation period, other factors such as cyclosporin A have to be considered for hyperlipidemia in renal transplant recipients. The observation on LDL-cholesterol being highest 18 months after transplantation suggests lipid lowering interventions to be indicated and might improve long-term outcome in renal transplant recipients. PMID- 9408683 TI - Effect of early cyclosporine levels on kidney allograft rejection. AB - Acute rejection is the greatest risk factor for development of biopsy-proven chronic rejection and late kidney allograft loss. We previously noted that low cyclosporine (CsA) levels were a risk factor for early acute rejection in pediatric recipients (1). In our current study, we used logistic regression to identify risk factors for acute rejection in 726 adult kidney transplant recipients on triple therapy (prednisone, azathioprine, CsA). Variables considered for logistic regression analysis were donor organ source (cadaver vs. living), degree of HLA mismatch (1 to 6 vs. 0 antigen mismatch), transplant number (primary vs. retransplant), CsA levels (< 125 vs. > or = 125 ng/ml, < 150 ng/ml vs. > or = 150 ng/ml, and < 175 vs. > or = 175 ng/ml), and acute rejection episodes (0 vs. > or = 1). Of 726 recipients, 401 (55%) received cadaver kidneys; 325 (45%), living related. Overall, 572 (79%) had a primary transplant; 154 (21%), a retransplant. The vast majority of acute rejection episodes occurred within the first 2 months posttransplant; 68% of recipients had no acute rejection episodes by 2 months and 58% had none by 60 months posttransplant. Logistic regression analysis revealed that a cadaver donor kidney (vs. living) (p = 0.004), a 1 to 6 antigen mismatch (vs. 0 mismatch) (p = 0.001), and CsA levels < 150 ng/ml (vs. > or = 150 ng/ml) correlated with biopsy-proven acute rejection. The correlation for CsA levels < 150 ng/ml (vs. > or = 150 ng/ml) held true for levels at 1 wk (p < 0.05), 1 month (p = 0.0001), 2 months (p = 0.01), and 3 months (p = 0.02) posttransplant. Similar correlation was found for CsA levels < 125 ng/ml (vs. > or = 125 ng/ml) and < 175 ng/ml (vs. > or = 175 ng/ml). Comparative analyses were made (by Chi-square) of acute and chronic rejection rates when recipients were divided into 3 groups by CsA level (< 125 ng/ml, > or = 125 to < 150 ng/ml, and > or 150 ng/ml). At each time point (1 wk, 2 wk, 1 month, 2 months, 3 months), CsA levels < 125 ng/ml (vs. > or = 125 to < 150 ng/ml and > or = 150 ng/ml) were associated with the greatest increased risk of acute rejection--for both cadaver and living related recipients (all p < 0.05). CsA levels < 125 ng/ml at each time point (1 wk, 2 wk, 1 month, 2 months, 3 months) were also associated with a significantly increased risk of chronic rejection (all p < 0.001). The incidence of both acute and chronic rejection was reduced in the group with CsA levels > or = 125 to < 150 ng/ml and further reduced in the > or = 150 ng/ml group. Our data indicate that maintaining CsA levels > or = 150 ng/ml as part of triple therapy reduces the incidence of both acute and chronic rejection. Because chronic rejection is the leading cause of late allograft loss, maintaining adequate CsA levels should improve long-term graft survival. Based on this analysis, we have modified our own immunosuppressive regimens to achieve higher CsA levels earlier posttransplant. PMID- 9408684 TI - IgM antibodies in renal transplantation. AB - A prospective final crossmatch with patient serum and donor lymphocytes using the complement-dependent cytotoxicity assay to identify any performed anti-donor antibody is required for kidney transplantation. The presence of pre-existing antibody may lead to hyperacute rejection of the transplanted kidney. Certain anti-donor antibodies have previously been shown to be ineffective in promoting hyperacute rejection, such as IgM autoantibodies and non-specific IgM lymphocytotoxic antibodies. In this report, we present evidence that IgM HLA alloantibody specific to the donor does not lead to hyperacute rejection and produces graft survival results equivalent to transplants with negative pre transplant final crossmatches. Forty-eight (48) of 402 patients transplanted over and 8 yr period were transplanted across a positive final crossmatch due to IgM antibodies alone. Three patients exhibited IgM autoantibodies and 26 patients demonstrated non-specific IgM antibodies to lymphocytes. In 15 patients, following a detailed serum screening analysis, a significant correlation (r > 0.9, p < 0.001) was observed between HLA Class I antigens and the presence of corresponding IgM alloantibodies. Five of these patients were subsequently transplanted despite a positive final crossmatch that was clearly demonstrated to be the result of IgM alloantibody to donor HLA Class I specificities. All of these patients continue to have graft function. These results suggest that hyperacute rejection is not mediated by any type of IgM antibody to donor lymphocytes and that kidney transplantation when only IgM antibody is present against donor lymphocytes represents a reasonable opportunity for a safe transplant and successful long-term graft survival. PMID- 9408685 TI - Remission of transplanted melanoma--clinical course and tumour cell characterisation. AB - The recipient of a cadaveric kidney was found to have donor melanoma within the graft together with metastatic spread. After cessation of immunosuppression, the kidney rejected and was removed. One month later there was both clinical and radiological evidence of remission and at autopsy 5 months later there was no histological evidence of melanoma. The outcome for recipients of other organs from the same donor was varied. Tumour cells expressed HLA class 1 antigens mismatched in the recipient and mRNA for the costimulator B7, and the cytokines GM-CSF, IL-1 alpha and beta. These characteristics may have been important in the host immunological response. PMID- 9408686 TI - Immunomagnetic bead cell isolates reduce kidney donor cold ischemia time, STAT crossmatch time and histocompatibility test cost. AB - The use of immunomagnetic beads for STAT cadaveric crossmatching for renal transplantation reduces test cost by 50%, laboratory work-up time by 50% and cold ischemia time by 4 h as compared with the nylon wool method of T- and B lymphocyte separation. PMID- 9408687 TI - Acute renal allograft rejection in patients with Epstein-Barr virus associated post-transplant lymphoproliferative disorder. AB - BACKGROUND: There is a reciprocal relationship between post-transplant lymphoproliferative disorder (PTLD) and rejection: aggressive treatment of rejection can result in PTLD, while treatment of PTLD by reducing immunosuppression can result in recrudescence of rejection. The literature on the relationship between PTLD and rejection episodes is limited. METHODS: The clinical course and outcome of rejection episodes occurring prior to and following a diagnosis of PTLD were studied in 20 renal transplant recipients. RESULTS: The diagnosis of PTLD was preceded by rejection in 12/20 (60%) patients. OKT3 treatment was associated with early onset PTLD, which involved the allograft in 6/7 patients (86%). The risk of rejection following reduced immunosuppression was 7/14 (50%). Post-PTLD rejection left untreated led to graft loss in 3 patients. The remaining 4 patients responded satisfactorily to anti-rejection therapy. CONCLUSIONS: Reduction of immunosuppression for PTLD is frequently, but not invariably, complicated by rejection. The clinical outcome of PTLD does not correlate with the occurrence or reversibility of rejection episodes. PMID- 9408689 TI - Donor biliary variations: an overlooked problem? AB - Documented causes of biliary complications following orthotopic liver transplantation have been related to technical imperfections or insufficient arterial supply. Although anatomical variations of the extrahepatic biliary system are not infrequent, neither their incidence, surgical management nor possible association with complications have been reported in liver transplantation. At our institution, the global incidence of biliary complications following 357 consecutive liver transplants performed in 324 patients over a 2-yr period was 15.4% (55/357). Anomalous donor extrahepatic ducts were verified in 10 cases (2.8%) and they were recognized intraoperatively, prior to biliary reconstruction, in 7 cases. Technical complications occurred in 1 of these 7 and in 3 other cases where the anomalous ducts were not identified until later in the postoperative period when serious clinical problems ensued. We herein present a description of these 10 cases, with reference to the techniques employed to manage the anatomical anomalies and to treat complications. As in any hepatobiliary procedure, awareness of possible variations of the extrahepatic biliary system, intraoperative identification of the anomalous ducts and appropriate tailoring of the surgical technique are advisable in order to avoid serious postoperative complications in liver transplantation. PMID- 9408688 TI - Elevation of cyclosporin A blood levels during carvedilol treatment in renal transplant patients. AB - In a consecutive study of 21 renal transplant patients suffering from chronic vascular rejection (CVR) we added the beta-receptor-blocking drug carvedilol to their regular medication. The purpose was to investigate possible pharmacokinetic interactions between carvedilol and cyclosporine (CsA), since carvedilol will soon be used in clinical trials in renal transplant patients with CVR. On the first day of the study the patients received 6.25 mg of carvedilol added to their daily medication. The dose was increased stepwise to 50 mg while the doses of other beta-blocking drugs were decreased. The goal was to exchange atenolol with carvedilol at a ratio of 2:1 when substituting atenolol with carvedilol. The patients' blood pressure was the final determinant of the dose of carvedilol. The trough levels of CsA were measured on days 1, 14, 30, 90 and 180. It was found that the blood levels of CsA increased when carvedilol was introduced. Thus, the doses of CsA had to be reduced in order to keep the blood levels within the therapeutic range. At 90 d, the daily doses of CsA had been reduced from 3.7 +/- 0.3 to 3.0 +/- 0.2 mg/kg BW (p < 0.001). The present results suggest an interaction between carvedilol and CsA that demands a 20% average reduction of CsA doses to maintain the CsA blood levels within the therapeutic range. However, the interaction shows a great interindividual variation, calling for careful monitoring of the CsA blood levels. PMID- 9408690 TI - Differential contribution of graft and recipient to perioperative TNF-alpha, IL-1 beta, IL-6 and IL-8 levels and correlation with early graft function in clinical liver transplantation. AB - Cytokines, produced by both the recipient and the newly vascularized allograft, are central mediators in the inflammatory response to allografted tissue. This study examines the relationship between pre- and intraoperative levels of TNF, IL 1, IL-6, and IL-8 and hepatic allograft function in the early postoperative period and also determines which cytokines are produced in a significant amount by the newly vascularized allograft. Baseline levels of IL-6 and IL-8 tended to be higher in patients with more advanced disease and showed an increase during the anhepatic period. TNF and IL-1 remained stable from baseline to anhepatic phase. IL-1 showed an increase from portal vein to effluent samples, suggesting that the graft has an important contribution to circulating IL-1 levels. Analysis of the data according to early graft performance revealed extremely high levels of effluent IL-1, IL-6 and IL-8, and the prolonged elevation of the latter two cytokines in patients with poor early graft function. Our findings demonstrate that sequential perioperative measurements of proinflammatory cytokines can be useful in monitoring graft function. PMID- 9408691 TI - Prophylactic use of low-dose urodilatin for prevention of renal impairment following liver transplantation: a randomized placebo-controlled study. AB - Many therapeutic measures have been employed to prevent or at least ameliorate postoperative renal impairment following liver transplantation. Recent clinical phase II studies have demonstrated that the new natriuretic peptide urodilatin has beneficial effects on renal function following heart and liver transplantation. The present study reports the first prospective randomized placebo-controlled trial of prophylactic urodilatin administration following liver transplantation. Seventy consecutive recipients of primary liver transplants were included in the study following randomization, and 33 patients continuously received urodilatin at a dose of 20 ng/kg/min for 7 d. The remaining 37 patients received a placebo infusion for the same time period. The course of serum creatinine and urea did not differ between the two groups nor did the daily urine production. However, the urodilatin group showed a higher preoperative median serum creatinine and a significant reduction on days 1 and 2, whereas this observation was not made in the placebo group. Furthermore, less furosemide was administered to the patients in the urodilatin group during the first 2 d. The incidence of postoperative hemodialysis and the number of treatments did not differ between the groups either (urodilatin group 4, vs. placebo group 6 and 22 for both groups, respectively). Side effects of the urodilatin therapy were not detected. The prophylactic low-dose urodilatin administration resulted in a trend towards amelioration of the renal function, but did not result in significant differences between the two experimental groups. Further studies, using higher doses, will be required to define the value of urodilatin for prevention of renal impairment after liver transplantation. PMID- 9408692 TI - Unexpectedly low immunocompetence in transplant patients on ketoconazole. AB - The P450 inhibitor ketoconazole may be used to decrease the dose, and therefore cost, of cyclosporine (CYA) by greatly decreasing the dose necessary to obtain therapeutic levels. However, the degree of immunosuppression produced using this drug regimen is not certain. We studied the immunocompetence of patients that had been started on ketoconazole to reduce the dose of CYA compared with patients treated conventionally. 95 assays were done in 64 patients including 6 assays in patients receiving low dose CYA plus ketoconazole. Immunocompetence was tested by measuring the mixed lymphocyte response using stimulators either non-depleted (ND) or depleted (D) of antigen presenting cells, based on the finding that CYA inhibits the response against D at a lower dose than against ND. Responses to ND/D ranged from +/+ through +/- to -/-. Normal individuals were always +/+. In conventionally treated patients with CYA the incidence of immunocompetence < or = +/- was 48%, whereas all patients on CYA + ketoconazole had an immunocompetence score < or = +/- (p = 0.03, chi 2). This degree of immunosuppression contrasted strikingly with the chemical levels, which for those on ketoconazole were in the low acceptable area (182.3 +/- 77.1 ng/ml range from 67 to 230 ng/ml). Therefore, patients using low-dose CYA plus ketoconazole to inhibit metabolism were more immunosuppressed than those receiving conventional CYA treatment, in spite of comparable CYA blood levels. If confirmed, this unexpectedly depressed immunocompetence in these patients would warrant caution in general regarding interpretation of trough blood levels in patients receiving CYA that are also being treated with agents that alter P450 activity. PMID- 9408693 TI - Beneficial effect of Sandoglobulin upon allograft survival in the pediatric renal transplant recipient. AB - The use of pooled immunoglobulin (IgG) has been shown to decrease panel reactive antibodies (PRA) in highly sensitized patients awaiting transplantation. IgG infusions have also been found effective for CMV prophylaxis. Analysis of 52 non highly sensitized children (ages 1-18) who received kidney transplants from May 1991 through January 1995 was undertaken to determine if the immunoglobulin administered for CMV prophylaxis effected allograft survival. Comparison of the "Sando Pos" group (those who received Sandoglobulin for CMV prophylaxis) to the "Sando Neg" group demonstrates a significantly improved allograft survival at 1, 2, and 3 yr post-transplantation. Despite the Sando Pos group being younger [7.3 +/- 1.3 yr vs. 10.7 +/- 0.9 yr; (mean +/- SEM) p < 0.05] allograft survival was 95%, 95% and 88% in the Sando Pos group vs. 88%, 79% and 79% in the Sando Neg group at 1, 2 and 3 yr, respectively (p < 0.01 at all three time points). It is concluded that the potential mechanism of the immunosuppressive benefit of Sandoglobulin is speculative but presumed to be upon inhibition of anti-HLA class I antibodies. We conclude that Sandoglobulin may not only be useful for CMV prophylaxis but also as an adjunct to routine immunosuppression. PMID- 9408694 TI - Adjuvant intrapleural amphotericin B therapy for pulmonary mucormycosis in a cardiac allograft recipient. AB - Infectious complications after heart transplantation remain a major cause of morbidity and mortality. While many viral, bacterial, and protozoal infections can be successfully treated, fungal infections continue to be challenging. Mucormycosis is a rare infection in heart transplant recipients; however, mortality is exceedingly high. We report a case of cavitary Rhizopus lung infection 2 months after cardiac transplantation. The infection was complicated by inadvertent exposure of the pleural cavity to the fungus during surgical resection. Therapy consisted of standard systemic amphotericin B, surgical excision, and for the first time, the use of adjuvant intrapleural amphotericin B. Cure was achieved with no clinical or radiological evidence of disease at 3 months follow-up. Rhizopus pulmonary infection is a rare complication of cardiac transplantation. Treatment consists of the triad of systemic anti-fungal therapy, surgical resection, and control of any underlying predisposing diseases. Adjuvant intrapleural amphotericin B use could also be considered in patients with fungal pneumonias and evidence of chest wall and/or pleural cavity involvement. PMID- 9408695 TI - Oral ganciclovir in pediatric transplant recipients: a pharmacokinetic study. AB - An oral formulation of ganciclovir (GAN) has been shown to be effective as prophylaxis of cytomegalovirus (CMV) after liver transplantation in an adult population. There are no data on the use or dose of oral GAN in pediatric transplant recipients. We evaluated the pharmacokinetics of oral GAN in a group of such patients. Nine patients (4 kidney and 5 liver transplant recipients, age 0.9-13 yr) were treated with the commercial formulation of oral GAN after transplant. All patients were considered at risk for CMV disease based on the use of anti-lymphocyte antibody (n = 5), and/or the use of a CMV positive organ in a CMV negative recipient (n = 5) or based on being a recipient of a liver transplant (n = 4). They received oral GAN for 84 +/- 29 d. All recipients had normal renal function as estimated by the Schwartz formula. While on a stable dose of oral GAN for at least 4 d (mean +/- SD 8.4 +/- 7, range 4-27 d), 1-ml serum samples were obtained before and at various times after dosing for the measurement of GAN levels. GAN levels were determined at a central laboratory by high-performance liquid chromatography. In 7 of the patients, a sufficient number of levels were obtained post-dosing to calculate the area under the curve using the linear trapezoidal rule. Cmin, the morning trough concentration, and Cmax, the peak concentration, were determined by inspection. Doses of oral GAN were increased if Cmin levels were less than 0.5-1.0 microgram/ml. Adequate levels of GAN were achieved in these patients at doses of 21.8-38.5 mg/kg or 568-990 mg/m2 every 8 h. There was a better correlation between the maximum GAN blood concentration and body surface area (R2 = 0.52, p = 0.008) than with body weight (R2 = 0.36, p = 0.04). Oral GAN was well tolerated in the 7 patients without evidence of leukopenia, thrombocytopenia, or nephrotoxicity. No CMV disease occurred, although one patient had probable CMV syndrome with the development CMV IgM antibodies. These data suggest that oral GAN may be safely administered to pediatric transplant recipients. With normal renal function, the dosing should be in the range 20-40 mg/kg or 500-700 mg/m2 q 8 h. Further data in children with impaired renal function is required. PMID- 9408696 TI - Patient death or renal graft loss within 3 yr of transplantation in a county hospital: importance of poor initial graft function. AB - Our purpose was to study the factors predictive of graft loss or patient death within 3 yr of renal transplant in a county population. 363 renal transplants performed between 1984 and 1991 at the Harbor-UCLA Medical Center were reviewed. There were 16 recipients (4%) who died and 76 (21%) who lost their grafts within 3 yr. Known causes of death were cardiac (44%), infection (38%) and malignancy (6%). There was a higher proportion of diabetics (44%), hypertensives (69%) and those with a past cardiac history (38%). There was also a high delayed graft function rate (56%). Of the 76 grafts that failed in the first 3 yr, 14% were due to noncompliance, 20% to grafts that never functioned, 9% to technical problems, 21% to acute rejection, 9% to recurrent disease and 26% to chronic rejection. The noncompliance group were younger (mean 34 yr), and more likely to be a first transplant (90%) and be non-white (82%). The rejection group was significant for high delayed graft function rate (84%) and frequent and early rejection (76% at least one rejection). We conclude that fair organ allocation requires a balance between equity and utility, but patient death or graft loss within 3 yr benefits neither the patient or society. Patient survival may be improved with diligent cardiac evaluation, particularly in older patients and diabetics. Patient education and monitoring for noncompliance is essential, particularly in young first-time recipients. Maneuvers to decrease delayed graft function are essential to improve long-term results. PMID- 9408697 TI - Effect of initial slow graft function on renal allograft rejection and survival. AB - Cadaver renal allografts with immediate excellent function have good long-term outcomes, while grafts with delayed function have been associated with an increased incidence of acute rejection (AR) and subsequent poor long-term graft survival. There is, however, an intermediate group with initial slow function whose outcome is not well defined. This group was examined by reviewing 510 patients that received primary cadaver transplants between 1/85 and 8/95. Recipients were grouped by initial function into: 1) those with immediate graft function (IGF), defined by serum creatinine (Cr) level < 3 mg/dl on post operative day (POD) 5 (n = 237); 2) those with slow graft function (SGF), defined by serum Cr > 3 mg/dl on POD 5 but no need for dialysis (n = 160); and 3) those with delayed graft function (DGF), defined by the need for dialysis in the first week post-transplant (n = 113). Five-year graft survivals were determined for each group by Kaplan-Meier methods and compared by a generalized Wilcoxon test. The incidence of AR in the first 6 months was significantly higher in those with SGF (40%) vs. those with IGF (30%) (p < 0.05); both groups had a lower incidence than those with DGF (47%) (p < 0.05). In the absence of AR, 5-yr graft survival was similar between the 3 groups, 94%, 97% and 92% for IGF, SGF and DGF respectively. In the presence of AR, 5-yr graft survival was significantly reduced in all groups, but most notably in those with SGF (51%) or DGF (53%), as compared to those with IGF (80%), (p < 0.001). We conclude that recipients with SGF, but no AR, have excellent outcomes, comparable to those with IGF. However, there is an increased incidence of early AR associated with SGF. In recipients with SGF or DGF, AR has a more profound detrimental effect on long-term graft survival than in the IGF group. Thus, recipients with SGF are at increased risk for AR with resultant poor long-term graft survival, and may need modified immunosuppressive protocols. PMID- 9408698 TI - Importance of decreased heart rate in predicting transplant coronary artery disease. AB - Studies in animals and humans have demonstrated that an increased heart rate is a predictor for the development of coronary atherosclerosis and overall cardiovascular mortality. In contrast, we have previously reported that the need for pacemaker implantation because of bradycardia in heart transplant recipients is associated with an increased prevalence of transplant coronary artery disease (TxCAD). Hence, the relevance of changes in heart rate to the development of TxCAD remains unclear. Intra-coronary ultrasound examinations (ICUS) were therefore analyzed in 130 heart transplant recipients (age 50 +/- 11 yr) studied at annual evaluations (3.7 +/- 3.0 yr after transplantation). Quantitative ultrasound measurements were obtained by calculating mean coronary artery intimal thickness (MIT) obtained by examination of the left anterior descending artery. The presence of TxCAD was defined as MIT > 0.3 mm. Resting heart rates (HR) were recorded with the patients in the supine position during routine echocardiography. Based on HR recordings, two groups were defined: group 1, HR below; or group 2, HR above the median. TxCAD was detected in 40% of the ICUS studies overall. The prevalence of TxCAD was higher in group 1 (49%) compared with group 2 (33%), p < 0.05. There was no significant difference in donor ischemic time or donor gender, recipient age, gender, body weight, CMV status, creatinine, total cholesterol, use of lipid lowering drugs or diltiazem. Donor age and use of beta-blockers were higher in group 1 compared with group 2 (29 +/- 10 vs. 25 +/- 9 yr, and 15% vs. 5%, for donor age and beta-blocker use, respectively). By multivariate regression analysis only donor age and years after transplantation were independently correlated with TxCAD. After excluding patients taking beta-blockers and diltiazem, the prevalence of CAD was still higher in group 1 (50%) vs. group 2 (34%). In conclusion, transplant coronary artery disease is more prevalent in patients with lower, rather than higher, heart rates. The reason for this is unclear, but may reflect impaired blood flow to the sinoatrial node. PMID- 9408699 TI - Efficacy of oral ganciclovir in prevention of cytomegalovirus infection in post kidney transplant patients. AB - Unlike parenteral gancicovir, the efficacy of oral ganciclovir in the prevention and treatment of cytomegalovirus (CMV) infection in kidney transplantation has not been well documented. This study prospectively evaluated the episodes of CMV infection within the first nine months after transplantation in renal transplant recipients treated prophylactically with oral ganciclovir (750 mg twice a day) over a period of 3 months (oral ganciclovir, N = 22), compared with patients randomly assigned as controls (controls, N = 22) who did not receive any anti viral prophylaxis. Diagnosis of CMV infection at presentation included serological determination of CMV-specific immunoglobulin M antibodies, CMV immunofluorescence assay [standard culture tube and shell vial] (blood) and virus isolation (urine and tissue). CMV infection occurred in one patient (5%) in the oral ganciclovir group and 6 patients (27%) in the control group (p < 0.05). The episodes of biopsy proven allograft rejections were 5% (1/21) and 18% (4/22) in the oral ganciclovir and control groups, respectively. Except for one, none of these patients developed CMV infection either before or after rejection(s). Controlling for the reason (induction or treatment of rejection) for using cytolytic antibody therapies, we found that prophylactic oral ganciclovir was protective against CMV infection (adjusted risk reduction 0.83; 95% confidence interval, 0.33-0.98; p < 0.05). Neither, the CMV status of donors and recipients nor the treatment for acute rejection had any significant impact on the occurrence of CMV infections. Our results show that oral ganciclovir is an effective and well tolerated therapy in the prevention of CMV infection in renal transplant patients. PMID- 9408700 TI - Comparative study of the effects of two once-a-month injectable steroidal contraceptives (Mesigyna and Cyclofem) on lipid and lipoprotein metabolism. United Nations Development Programme/United Nations Population Fund/World Health Organization/World Bank, Special Programme of Research, Development and Research Training in Human Reproduction. AB - A 1-year longitudinal multicenter study was undertaken to monitor the effects on lipid metabolism of two once-a-month injectable contraceptive preparations, Mesigyna (50 mg norethisterone enanthate and 5 mg estradiol valerate and Cyclofem (25 mg medroxyprogesterone acetate and 5 mg estradiol cypionate). A total of 357 volunteers from four centers (Hangzhou, Havana, Jakarta, and Shanghai) were monitored during one pretreatment cycle, the third and ninth injection intervals, and during one post-treatment cycle, approximately 3 months after the 9-month period of treatment was completed. With both preparations, changes were observed at 3 months of use, remained essentially unchanged during treatment, and returned to pretreatment levels after discontinuation of use. With Mesigyna, changes consisted mostly of decreases in high density lipoprotein cholesterol (HDL-C, 10%), apolipoprotein AI (-9%), and triglycerides (-15%). With Cyclofem, changes were even smaller. With both, variations in lipid levels mirrored closely the pharmacokinetics of each preparation and the biphasic nature of the injection intervals. PMID- 9408701 TI - Depot medroxyprogesterone acetate. Patterns of use and reasons for discontinuation. AB - Little information is available from outside clinic settings about the acceptability of depot medroxyprogesterone acetate (DMPA, Depo-Provera) as an injectable contraceptive. In this national, population-based study, New Zealand women aged 25 to 54 years were selected at random from voter rolls. The 1864 subjects were interviewed by telephone after an initial approach by letter. More than 1 in 8 women (13.7%) had used DMPA at some time. The proportion was higher among Maori women and among those of lower income and education, but DMPA had been used by a substantial proportion of all socioeconomic groups. A quarter of all users reported receiving only a single injection of DMPA, and only 53% had used this method for a total of 12 months or more. Only 5 (1.6%) of discontinuations were attributed to contraceptive failure; this corresponds to a contraceptive failure rate of 0.9 per 100 woman-years. Side effects were given as the most common reasons for stopping, with menstrual disturbances and weight gain being cited most often. Other reasons for stopping included no further need for contraception or doubts about the appropriateness of DMPA. In this developed country population, DMPA is widely used for short periods but its acceptability is limited by the occurrence of side effects. PMID- 9408702 TI - Results of a pilot study of the time to azoospermia after vasectomy in Mexico City. AB - In a joint pilot study by Family Health International (FHI), AVSC International, and the Instituto Mexicano del Seguro Social (IMSS), information was gathered on the determinants of azoospermia following vasectomy on 38 healthy men who chose vasectomy for contraception. The time and number of ejaculations associated with loss of sperm motility and loss of sperm eosin vital staining were also evaluated. "Azoospermia" was defined on the basis of two consecutive azoospermic semen samples collected at least 3 days apart. The single decrement life table method was used to calculate weekly gross cumulative life table rates for the time to azoospermia, zero motility, and zero viability. The Kaplan-Meier method was used to calculate the product-limit estimates of the cumulative rates for the total number of ejaculations to azoospermia, 0% motility and 0% viability. The median time to azoospermia was 10 weeks and at the end of week 20, the life table rate (+/- standard error) was 93.0 (+/- 6.30) weeks/100 men. The median number of ejaculations to azoospermia was between 25 and 30, and the cumulative rate (+/- standard error) at 60 ejaculations reached 94.5 (+/- 5.16)/100 men. PMID- 9408703 TI - Fertility regulation in nursing women. IX. Contraceptive performance, duration of lactation, infant growth, and bleeding patterns during use of progesterone vaginal rings, progestin-only pills, Norplant implants, and Copper T 380-A intrauterine devices. AB - This study evaluated the performance of progesterone vaginal rings (n = 187), progestin-only pills (n = 117), Norplant implants (n = 120), and Copper T 380-A intrauterine devices (n = 122) in lactating women. Contraceptive efficacy, bleeding pattern, and influence of the method upon breastfeeding duration and infant growth were compared with those of untreated women (n = 236) who relied on lactational infertility. Participants were healthy, 18 to 38 years, had had a normal delivery, and were intending to breastfeed for as long as possible. Contraceptives were initiated at day 57 +/- 3 postpartum. Results are reported for the first year of use. All methods were highly effective, with pregnancy rates below 1%. None affected breastfeeding performance or the rate of infant growth. Users of the progestin-only methods experienced a period of lactational amenorrhea 4 to 5 months longer than did users of Copper T or untreated women. More than half of the women in each contraceptive group reported a bleeding in the first month after treatment initiation, which was not considered in the calculation of the duration of amenorrhea. Prolonged or frequent bleedings were infrequent. The proportion of bleedings lasting more than 10 days ranged from 0 in the progestin-only pills group to 7% in the Norplant implants group. The four methods, initiated around the eighth postpartum week, provided effective contraception with no negative effects upon lactation or infant growth and without the bleeding problems associated with their use in nonlactating women. PMID- 9408705 TI - The contraceptive choice for a Wilson's disease patient with chronic liver disease. AB - Preserved fertility status is frequently encountered in patients with Wilson's disease, and contraceptive counseling may, therefore, be a relevant issue. Yet, several contraceptive methods can adversely influence the hepatic function, and the efficacy of others may be affected by the liver disease. We describe a patient with Wilson's disease manifested by cirrhosis, portal hypertension, and bleeding esophageal varices who underwent termination of pregnancy at 9 weeks' gestation. Following the procedure, contraceptive advice was sought in order to postpone conception until portal hypertension was controlled and hepatic function improved. Intramuscular depot medroxyprogesterone acetate was administered and tolerated well by the patient. A detailed discussion of the contraceptive options for patients with chronic liver disease, in general, and Wilson's disease, in particular, follows the case report. PMID- 9408704 TI - Effect of different insertion regimens on side effects with a combination contraceptive vaginal ring. AB - The study population included 159 women aged 18 to 37 years requiring contraception (60 in Sydney, 99 in Los Angeles). The design consisted of a 6 month study of a contraceptive vaginal ring (CVR) releasing 20 micrograms ethinyl estradiol and 1 mg of norethindrone acetate daily over two successive cycles with a new ring on each of three different insertion regimens. A total of 831 woman months of exposure were recorded, and 129 women completed the study. The overall incidence of side effects was similar in the two centers and differences between the insertion regimens were not observed. Transient nausea following insertion of a new ring (mainly lasting 0.5 to 48 h) was the most prominent side effect, with no significant difference between the insertion regimens (although the incidence tended to be highest with bedtime insertion [34%] and lowest with early evening insertion and temporary removal during the first night [27%]). Transient vomiting occurred in about 10% of women in the first 24 h after first insertion of a new ring. The incidence and severity of nausea was greatly reduced in cycle 2 with each regimen (6% to 9% of women). Nausea could be prevented by overnight soaking of the ring in water before use. Other side effects such as headache, dizziness, uterine cramps, breakthrough bleeding, weight gain, and ring expulsion occurred with similarly low frequency in all three insertion groups. One pregnancy occurred with probable ovulation between cycles 5 and 6, the only pregnancy recorded to date in studies with this ring. The study demonstrated that this effective and generally well tolerated CVR causes a relatively high incidence of transient nausea after insertion of a new ring, which is probably due to accumulation of ethinyl estradiol on the ring surface during storage. Acceptability is still high, and this particular CVR merits further development. PMID- 9408707 TI - Gossypol-induced hypokalemia and role of exogenous potassium salt supplementation when used as an antispermatogenic agent in male langur monkey. AB - In our earlier study, we have observed that hypokalemia in langur monkeys, following gossypol acetic acid (GAA) treatment (5 mg dose level) when used as an antispermatogenic agent, and potassium salt supplementation partially maintained body potassium level of the animals. The aims of the present investigation was to confirm further occurrence of hypokalemia in the monkey (comparatively at two higher dose levels) and the role of potassium salt in preventing occurrence of gossypol-induced hypokalemia. Highly purified gossypol acetic acid alone at two dose levels (7.5 and 10 mg/animal/day; oral) and in combination with potassium chloride (0.50 and 0.75 mg/animal/ day; oral) was given for 180 days. Treatment with gossypol alone as well as with the supplementation of potassium salt resulted in severe oligospermia and azoospermia. Animals receiving gossypol alone showed significant potassium deficiency with signs of fatigue at both dose levels. Enhanced potassium loss through urine was found in potassium-deficient animals, whereas animals receiving gossypol acetic acid plus potassium salt showed normal serum potassium with a less significant increase in urine potassium level during treatment phases. Other parameters of the body remained within normal range except gradual and significant elevation in serum transaminases activity. The animals gradually returned to normalcy following 150 and 180 days of termination of the treatment. PMID- 9408706 TI - Phase II clinical trial of a vas deferens injectable contraceptive for the male. AB - Following up on an earlier clinical trial demonstrating the safety of an intra vas deferens injection of a contraceptive drug named Risug, comprised of styrene maleic anhydride (SMA) in a solvent vehicle of dimethylsulphoxide (DMSO), a study to assess the contraceptive effectiveness of a specific dose (60 mg) of SMA bilaterally was planned and implemented. Male subjects and their wives with normal reproductive profiles were the volunteer subjects. The wives were not using any contraceptives. The results reconfirm the safety and show that for a period of at least 1 year, the treatment leads to azoospermia in the male and gives pregnancy protection. PMID- 9408708 TI - Use of norethisterone and estradiol in mini doses as a contraceptive in the male. Efficacy studies in the adult male bonnet monkey (Macaca radiata). AB - Administration of norethisterone (NET) or NET + estradiol benzoate using an Alzet minipump or as once-a-month intramuscular injection of their depot forms, NET enanthate (NET-EN) and estradiol valerate (E-val), resulted in azoospermia in all monkeys (n = 13) within 60 to 150 days of treatment. Although addition of depot form of testosterone (T, 20 mg/month) to the regimen restored the behavioral response typical of a normal male, it did not reverse the azoospermic state. Serum T (heightened nocturnal) levels were significantly reduced (> 85%, p < 0.001) in all the treated groups. Evidence for blockade in spermatogenesis following treatment was obtained by DNA flow cytometry. Following withdrawal of treatment, the T level was restored to normalcy within 15 days but 120 days more were required for the animals to exhibit normal sperm counts. In conclusion, the efficacy of once-a-month injection of relatively low doses of NET-EN + E-Val to bring about azoospermia in monkeys, in a relatively short time, has been demonstrated. As the results are uniform and reproducible, it appears desirable that this steroid regimen be tested in man for its contraceptive efficacy. PMID- 9408709 TI - Fertility impairment after mifepristone treatment to rats at proestrus. Actions on the hypothalamic-hypophyseal-ovarian axis. AB - Accumulated evidence indicates that the antigestagen mifepristone affects the reproductive axis acting on hypothalamic, pituitary, ovarian, and uterine tissues. The purpose of this study was to further investigate which reproductive functions are impaired by the antagonist, critically compromising the reproductive process, leading to unsuccessful pregnancy. Circulating pituitary and ovarian hormones, sexual receptivity, ovulation, and implantation rates were studied in cycling rats receiving a single dose of mifepristone (1 or 10 mg/kg subcutaneously) at 12:00 proestrus, before luteinizing hormone (LH) stimulation of the ovulatory process. Mifepristone-treated rats had decreased preovulatory surges of LH and prolactin (PRL), and hypersecretion of LH, PRL, and progesterone at estrus. The sexual receptivity was dramatically affected by the antagonist as indicated by the profound decrease in the lordosis response evaluated on the night of proestrus. The number of ovulating animals and the number of oocytes recovered from the oviduct on the morning of estrus were not affected by mifepristone. The low number of rats that succeeded in mating with potent males became pregnant. However, they delivered an average of only two pups at parturition, indicating a failure in the implantation of the fertilized ova, as ovulation was not affected by the antagonist at the dose used. We conclude that a dramatic inhibition of the sexual receptivity and unsuccessful implantation, preceded by a reduction on LH and PRL secretion, are the major components leading to fertility impairment after a single dose of mifepristone administered before the preovulatory surge of LH. PMID- 9408710 TI - Surgical procedures and devices should be evaluated in the same way as medical therapy. AB - This paper is a personal essay that starts and ends with the message that surgical procedures and devices should be evaluated in the same way as medical therapies, namely, by randomized clinical trials (RCTs). I discuss, with particular attention to surgical procedures and devices, the objections raised against RCTs in medical decision-making, a schema for utilizing the traditional phases of RCTs in the evaluation of surgical procedures and devices, the importance of RCTs to FDA approval, financial compensation, and health care costs, the impact of RCTs on clinical practice, the role of RCTs in academia, teaching, and research, and the surgeon's obligation to participate in a leadership role in RCTs. The belief is expressed that national funding of health care should mandate allocations for RCTs and that such expenditures, as well as the spending of health care dollars on the basis of the outcomes of such trials, will not only improve patient management in the shortest time but will eventually reduce health care costs. The RCT, by selecting effective and safe surgical procedures and devices, as well as diets and drugs, is the best means science has to assess the validity of patient management. PMID- 9408711 TI - Influence of study goals on study design and execution. AB - From the viewpoint of a clinician who makes recommendations to patients about choosing from the multiple possible management schemes, quantitative information derived from statistical analyses of observational studies is useful. Although random assignment of therapy is optimal, appropriately performed studies in which therapy has been nonrandomly "assigned" are considered acceptable, albeit occasionally with limitations in inferences. The analyses are considered most useful when they generate multivariable equations suitable for predicting time related outcomes in individual patients. Graphic presentations improve communication with patients and facilitate truly informed consent. PMID- 9408712 TI - Observational epidemiology is the preferred means of evaluating effects of behavioral and lifestyle modification. AB - In evaluating the health effects of behavioral change, randomized trials have the clear advantage of being able to provide unbiased and unconfounded estimates. However, in practice, many difficulties loom that limit the utility of trials. These include the inability to assess adverse behavior (except indirectly), the often unrealistic requirement for very prolonged adherence--perhaps for decades- to the randomized assignment, the requirement for huge numbers for precise estimates of effect (as opposed to simply a "significant result"), and the ethical problem of giving a placebo when an efficacious agent is available. We must be alert to feasible opportunities to test behavioral changes in clinical trials, but the many limitations mean that we will continue to rely on well conducted, large-scale observational studies for most of our evidence on the effects of behavioral change. PMID- 9408713 TI - Individual subject random assignment is the preferred means of evaluating behavioral lifestyle modification. AB - Of the three most important approaches to evaluating lifestyle and health outcomes--observational studies, individual subject random assignment clinical trials, and community random assignment clinical trials--individual subject random assignment clinical trials provide the most useful information and the most certain inferences. Observational studies are limited by the collection of information on association of lifestyle instead of change in lifestyle with health outcomes, as well as by individual lifestyle selection that may be associated with particular outcomes. Community randomized trials may be the best way to decide such public health policy issues as whether or not to add fluoride to a water supply or to use a community-wide anti-smoking program. The limited amount of individual-specific data collected in community randomized trials, difficulties in accounting for missing data, and problems in data analysis because people move into or out of communities that are under study limit the value of community randomized trials for advising individuals whether or not to embark on a lifestyle modification program. Clinical trials of pharmacologic agents have successfully addressed challenges to individual subject random assignment clinical trials of lifestyle modification, such as long duration of study, access to study intervention(s) by individuals not assigned them, and cost. PMID- 9408714 TI - The advantages of community-randomized trials for evaluating lifestyle modification. AB - Observational studies may provide suggestive evidence for the results of behavior change and lifestyle modification, but they do not replace randomized trials for comparing interventions. To obtain a valid comparison of competing intervention strategies, randomized trials of adequate size are the recommended approach. Randomization avoids bias, achieves balance (on average) of both known and unknown predictive factors between intervention and comparison groups, and provides the basis of statistical tests. The value of randomization is as relevant when investigating community interventions as it is for studies that are directed at individuals. Randomization by group is less efficient statistically than randomization by individual, but there are reasons why randomization by group (such as community) may be chosen, including feasibility of delivery of the intervention, political and administrative considerations, avoiding contamination between individuals allocated to competing interventions, and the very nature of the intervention. One example is the Community Intervention Trial for Smoking Cessation (COMMIT), which involved 11 matched pairs of communities and randomized within these pairs to active community-level intervention versus comparison. For analysis of results, community-level permutation tests (and corresponding test based confidence intervals) can be designed based on the randomization distribution. The advantages of this approach are that it is robust, and the unit of randomization is the unit of analysis, yet it can incorporate individual-level covariates. Such covariates can play a role in imputation for missing values, adjustment for imbalances, and separate analyses in demographic subsets (with appropriate tests for interaction). A community-randomized trial can investigate a multichannel community-based approach to lifestyle modification, thus providing generalizability coupled with a rigorous evaluation of the intervention. PMID- 9408715 TI - Clinical perspectives on the use of composite endpoints. AB - Although mortality is the most important endpoint in evaluating new regimens, insistence on its use as the only endpoint in clinical trials can require that thousands of patients be studied. Accordingly, composite endpoints have been increasingly used to increase the overall event rate and thereby reduce the number of patients needed for the trial. For use as part of composite endpoint, nonfatal endpoints should be clinically meaningful, i.e., related to an adverse subsequent prognosis. In acute myocardial infarction (MI), several intermediate endpoints in the well-described pathophysiology of acute MI have been correlated with an adverse long-term outcome: recurrent MI, new onset congestive heart failure or cardiogenic shock, left ventricular dysfunction, large infarct size, and failure to achieve early patency of the infarct-related artery. Furthermore, in acute MI, new therapies that improve these nonfatal endpoints also improve mortality, thereby validating this approach. Once this link is established, such nonfatal endpoints can be validly used in evaluating new therapies. Note, however, if this link has not been made (that mortality is reduced when there is a reduction in the nonfatal endpoint), as in the case of suppression of ventricular premature complexes with antiarrhythmic therapy, the nonfatal endpoint cannot be used validly. Thus, appropriately designed and validated composite endpoints can provide a valid means of testing new treatments in a smaller trial than one using mortality alone. Their use should allow testing of a greater number of new regimens, thereby allowing more rapid progress toward improving the clinical outcome of patients. PMID- 9408716 TI - Clinical trials with multiple outcomes: a statistical perspective on their design, analysis, and interpretation. AB - This article tackles both practical and statistical issues in the handling of multiple outcomes in clinical trials, with relevance to trial design, analysis, and reporting. Specific topics illustrated by examples include: the advantage of prespecifying priorities amongst outcomes and analyses, corrections for multiple significance testing and their limited value, problems with adverse event data, the use of a single global test of significance for clinically related outcomes, the use of a combined outcome for clinical event data, and the value of exploring interrelationships amongst outcomes. The problems in handling multiple outcomes are enhanced by trials being too small, dichotomous attitudes (is the trial "positive" or not?), obsession with p-values, and the manipulative instincts of human nature. While predeclarations of priorities in analysis and reporting of multiple outcomes are important in suppressing distortive claims, it would be unfortunate if too inflexible an approach suppressed unpredictable findings from being seriously considered. PMID- 9408717 TI - Secondary endpoints cannot be validly analyzed if the primary endpoint does not demonstrate clear statistical significance. AB - There is lack of consensus surrounding the interpretation of observed treatment effects for secondary clinical endpoints when the primary endpoint for which the clinical trial was initially designed does not meet the objective of a demonstrated effect. We provide some arguments to support caution in making inferences for secondary endpoints in this situation. We examine the definitions of primary and secondary endpoints within the context of a hypothesis-testing framework for multiple endpoints, and we address the relationship of the correlation structure of these endpoints and the statistical adjustments needed to preserve experiment-wise type I error for a valid inference. We also address the hypothesis-testing framework and the estimation framework for valid inference, focusing on the interpretation of p-values associated with differentially powered hypothesis tests for each endpoint to detect an important clinical effect. We point out the limitations on the strength of evidence (and quantification of uncertainty) for a secondary endpoint effect that can be derived from only one study and introduce the likelihood of replication of the finding in another study of identical size and design as a useful concept to guide this interpretation. PMID- 9408718 TI - Secondary endpoints can be validly analyzed, even if the primary endpoint does not provide clear statistical significance. AB - Response variables from clinical trials are often divided into those considered primary and those considered secondary to the purposes of the study. If the difference between treatment groups on primary outcomes is not significant, the interpretation of significant differences in secondary response variables may be difficult because of multiple comparisons. It is proposed that the comparison on the primary outcome use a significance level of alpha and that the secondary endpoints be evaluated using a significance level alpha/(k + 1), where k is the number of secondary outcomes to be evaluated. Examples from recently completed clinical trials illustrate the use of this rule. PMID- 9408719 TI - Strengths and limitations of meta-analysis: larger studies may be more reliable. AB - Meta-analysis of randomized controlled trials combines information from independent studies that address a similar question to provide more reliable estimates of treatment effects. At the present time, the methodology and usefulness of meta-analysis is under scrutiny. In the first part of this paper, we summarize the limitations of meta-analysis and make suggestions for improvements. In the second part, we illustrate strengths and limitations using examples of meta-analyses and subsequent large trials that address the same question. We develop the hypothesis that the size of the meta-analysis may be a useful measure of reliability. Small meta-analyses (i.e., those with less than 200 outcome events) may only be useful for summarizing the available information and generating hypotheses for future research. The results of small meta-analyses should be regarded with caution, even if the p value shows extreme statistical significance. Larger meta-analyses (i.e., those with several hundred events) are likely to be more reliable and may be clinically useful. Well-conducted meta analyses of large trials using individual patient data may provide the best estimates of treatment effects in the cohort overall and in clinically important subgroups. PMID- 9408720 TI - Cumulating evidence from randomized trials: utilizing sequential monitoring boundaries for cumulative meta-analysis. AB - We propose the adaptation of classical monitoring boundaries for use in cumulative meta-analysis as guidelines for deciding when accumulating evidence is statistically significant and medically convincing. The interpretation of information from a randomized controlled trial is compared with that from a meta analysis. The concept of optimal information size for a meta-analysis is developed and used to adapt monitoring boundaries to cumulative meta-analysis. PMID- 9408721 TI - Meta-analysis of randomized trials: looking back and looking ahead. AB - Meta-analyses as currently practiced are usually retrospective. They can be made more rigorous by developing a protocol that incorporates prospectively the elements that are usually necessary in a well-designed trial. Meta-analysis and large trials are complementary. Meta-analysis of small trials is useful in generating the hypotheses and assisting in the design of the large trials that are needed. Once the large trials have been completed, they could be brought together within the framework of a meta-analysis to estimate the overall treatment effect with greater confidence and to explore the effects in various subgroups. This article explores the value and limitations of meta-analyses and suggests ways of improving their conduct and interpretation. PMID- 9408722 TI - When are placebo-controlled trials no longer appropriate? AB - This paper presents a standard for assessing the validity of placebo-controlled trials in circumstances in which such trials might be unjustly denying appropriate therapies to members of the control group. This standard categorizes the types of risks that can or cannot be imposed upon consenting research subjects in such control groups. The paper also shows how needed research can be conducted while respecting the proposed ethical standard. Both the problem and the proposed standard are illustrated by reference to the major trials of the thrombolytic agents. PMID- 9408723 TI - When are clinical trials of a given agent vs. placebo no longer appropriate or feasible? AB - In several situations, placebo-controlled trials may be difficult to conduct. This article discusses the reasons for such difficulty--principally, a belief that the effect of a therapy has already been established or the existence of another therapy already proved effective. In the latter case, an active control equivalence trial may be considered, but this design has important deficiencies in many situations, notably, the lack of assurance of assay sensitivity. PMID- 9408724 TI - Approaches to informed consent. AB - Informed consent (IC) is an indicator, or a pivotal point, in broader and more fundamental questions dealing with the way clinical experimentation and, more specifically, randomized controlled trials (RCTs) relate to routine clinical practice; the rules that characterize the doctor-patient relationship; the self perception of medicine with respect to its capacity, duty, and autonomy in the production of new knowledge; and the role of medicine in society. The asymmetry of knowledge and power that characterizes the usual relationship between care providers and patients does not resolve when something experimental enters the relationship. The real world of clinical investigation is not uniformly distinct from clinical practice. Experimentation is more appropriately considered a continuum with respect to appropriate or recommended care. Fundamental patient rights come first and are more binding than compliance with procedures and regulations. The view that IC is the most important component of the "ethical" aspects of experimentation is highly misleading. The responsibility to foster well-informed decisions shapes the contents, the timing, the validity, and the credibility of IC. Documented, evaluable decisions are the natural substitute for individual IC when the patient is not able to handle information autonomously. Positive examples of IC practices and approaches suggest that IC may be important in improving the way medicine responds to its responsibilities and communicates with society. PMID- 9408725 TI - Informed consent: protection or obstacle? Some emerging issues. AB - There is widespread consensus on the need for informed consent procedures in medical research. Nevertheless, aspects of the informed consent process remain controversial, and innovative approaches to research may raise new issues and concerns. The randomized consent design for clinical trials, proposed by Zelen (N Engl J Med 1979; 300:1242-1245), permitted physicians to randomize patients without consent, then obtain informed consent from only those patients randomized to the experimental (as opposed to the standard treatment) arm. More recently, the proposal has been made to allow waiver of informed consent for study of patients in emergency circumstances who may be temporarily incapable of providing such consent, and for whom no family member is immediately available to give a "proxy" consent (Biros M.H. et al. JAMA 1995; 273:1283-1287). The medical community and federal regulatory policy have responded differently to these proposals. PMID- 9408726 TI - Distinctions between fraud, bias, errors, misunderstanding, and incompetence. AB - Randomized clinical trials are challenging not only in their design and analysis, but in their conduct as well. Despite the best intentions and efforts, problems often arise in the conduct of trials, including errors, misunderstandings, and bias. In some instances, key players in a trial may discover that they are not able or competent to meet requirements of the study. In a few cases, fraudulent activity occurs. While none of these problems is desirable, randomized clinical trials are usually found sufficiently robust by many key individuals to produce valid results. Other problems are not tolerable. Confusion may arise among scientists, scientific and lay press, and the public about the distinctions between these areas and their implications. We shall try to define these problems and illustrate their impact through a series of examples. PMID- 9408727 TI - Developing systems for cost-effective auditing of clinical trials. AB - Auditing a clinical trial is a complex process designed to ensure that the trial will provide a reliable answer to the question being posed. Traditional auditing methods are expensive, and escalate the cost of clinical trials. This paper describes approaches to cost-effective monitoring of clinical trials, such as integrating them with clinical practice and focusing the data being collected. Sampling methods for source documentation can be used to eliminate costs incurred by reviewing every record. These measures, coupled with prospective clinical judgment about areas of concern in the conduct of trials, can reduce complications and costs without sacrificing quality. PMID- 9408728 TI - Food materials adhesion: a review. AB - This article reviews the various theories of adhesion mechanism and, more specifically, studies concerning foodstuffs adhesion to industrial equipment and packaging surfaces. Adhesion is governed by mechanical interlocking, wetting, electrostatic and chemical forces, and diffusion. Direct conclusions about the validity of one of these theories were seldom made in the empirical studies reviewed. The different food adhesion determination methods were detailed: direct observations, evaluations (weighting, UV absorbance, and adhesive loss), adhesion strength measurements, and indirect measurements via the wetting theory (tilted plane method, contact angle, and surface tension). The importance of proteins, product rheological properties, solid surface rugosity, and wetting phenomena in many adhesion cases is highlighted. Conclusions were made that fundamental mechanisms of food-contact surfaces interactions still need to be investigated to improve understanding in the science of food materials. PMID- 9408729 TI - Variability in the intercomparison of food carotenoid content data: a user's point of view. AB - The availability of reliable information on food composition is essential both for the evaluation of diet and for nutritional research to relate diet to health or disease. In this article, we compare the total and individual carotenoid contents and the retinol equivalents in fruits and vegetables reported in several food composition tables and HPLC studies. The impact of the variability in carotenoid intake was evaluated on the basis of Spanish National Consumption Statistics and on the values for a standard diet. We identify, from a user's point of view, errors concerning identification of the items and the terms used to refer to the compounds. Food composition tables overestimate (by 2 to 48%) the retinol equivalent intake, whereas they underestimate (by 30 to 50%) the total carotenoid intake according to HPLC data. We study the effect of these main sources of error, their impact on dietary assessment and on the classification of relevant contributors, and the possible consequences with respect to proper diet in terms of nutritional assessment and epidemiological studies. Given the different dietary habits among populations and the fact that certain items may be over- or underestimated in databases, the use of a single database may be misleading as to the rate of carotenoid consumption and the "true" nutrient intake in a given population, thus weakening the reliability of the study and resulting in erroneous conclusions. PMID- 9408730 TI - Regulating the use of degraded oil/fat in deep-fat/oil food frying. AB - During frying, the degradation of oil produces harmful compounds. Improper monitoring of oil-discard times in restaurants either risks the public health or causes financial losses to industries. Measuring the oil quality is a complex problem and an online sensor is needed. The process of frying reviewed includes moisture, heat and fat/oil transfer, crust formation and various structural, textural and chemical changes in the product, and degradation of frying medium. Some of the European nations and the U.S. have specific regulations against the use of deteriorated frying oils. Due to the absence of a suitable online frying oil quality sensor for restaurant situations, it is difficult to implement any regulation against the use of deteriorated frying oil. Based on various regulations, a model regulation to increase the safety and quality of fried foods is discussed. Background and requirements for developing an online sensor to measure frying oil quality are discussed. Other related areas reviewed in this article are factors affecting oil penetration and absorption by the food, surfactant theory of frying, analytical indices, quick tests and acceptability of frying oil. PMID- 9408731 TI - Isolation, purification, and alteration of some functional groups of major milk proteins: a review. AB - This review covers selected methods of isolation and purification of mainly alpha s-casein, beta-casein, kappa-casein, beta-lactoglobulin, and alpha-lactalbumin. Selected methods of alteration of some functional groups of these proteins also were reviewed. Isolation and purification of milk proteins per se are methods of modifying the individual milk proteins. Gram quantities of these proteins can now be purified in a relatively short time using ion-exchange resins. Due to the prominent use of non-food-grade reagents in the procedures for preparation of these milk proteins, individual proteins are not maximally utilized for the manufacture of food/feed and pharmaceutical products. Therefore, intensive research efforts are needed to obviate the problems associated with underutilization of milk proteins. PMID- 9408732 TI - Pulmonary retention and clearance of inhaled biopersistent aerosol particles: data-reducing interpolation models and models of physiologically based systems--a review of recent progress and remaining problems. AB - During the last 40 years, most models of long-term clearance and retention of biopersistent particles in the pulmonary region of the lung were phenomenologically oriented and accounted for only a small portion of the growing insight into lung dynamics by pulmologists, histologists, and biochemists. In this review, theoretical developments of modeling pulmonary dynamics for biopersistent particles during or after inhalation exposure are discussed. Several characteristic examples are given of the present state of the art. Most of the models presently in use are pragmatical compartmental models with a single compartment for the pulmonary region. They relate to observed data and facilitate an interpolation within the range covered by observation. Occasionally, these models are unjustifiably used for extrapolations in efforts to derive hypothetical risk assessments. Modeling efforts aiming at models of physiologically based pulmonary systems with a potential for extrapolations are not common and were published only during the last decade. Of this kind of approach, the review covers four examples. Promising progress has been made, but scarcity of supporting experimental data slows validation and extension. The two most recent model developments are based on a hypothesis by P.E. Morrow. According to Morrow, alveolar clearance is accomplished by mobile alveolar macrophages after phagocytosis of particles on the alveolar surface. The macrophage mobility, however, and thus the efficiency of the transport to the mucociliary escalator of the tracheobronchial tract will eventually decline towards total loss of mobility after the particle burden of the macrophages exceeds a critical value. The POCK model has been evaluated for a variety of chronic and subchronic rat exposure studies with noncytotoxic aerosols and gave good simulation results. The model by Tran et al. appears to be still in the developing stage of facilitating simulations for cytotoxic aerosols, but the combination of both model approaches seems to be a sound route of future efforts. PMID- 9408733 TI - A review of epidemiologic studies of triazine herbicides and cancer. AB - We evaluated epidemiologic evidence pertaining to the human carcinogenic potential of triazine herbicides in general and of atrazine, the most common triazine. Cancers for which data are available included non-Hodgkin's lymphoma, Hodgkin's disease, leukemia, multiple myeloma, soft tissue sarcoma, colon cancer, and ovarian cancer. The investigations had methodologic limitations, including lack of in-depth exposure measurements and small numbers of subjects with heavy exposure and/or with many years since starting exposure, possibly required for the induction of cancer. The relation between triazines and non-Hodgkin's lymphoma has been assessed in four independent population-based case-control studies, reporting odds ratios ranging from 1.2 to 2.5. However, chance and/or confounding by other agricultural exposures may have produced these weak statistical associations. Furthermore, a pooled analysis of three of the case control studies and the combined analysis of two retrospective follow-up studies did not demonstrate the types of dose-response or induction time patterns that would be expected if triazines were causal factors. The epidemiologic data pertaining to Hodgkin's disease, leukemia, multiple myeloma, soft tissue sarcoma, colon cancer, and ovarian cancer were inadequate for determining whether associations with atrazine or triazines exist in humans. For each of these cancers, only one or two studies evaluating the relationship were available, and the results of the studies typically were imprecise. PMID- 9408734 TI - Albuminuria in non-insulin-dependent diabetes mellitus. Prevalence, causes, and consequences. PMID- 9408735 TI - Solution of methodological problems in prorenin measurement and investigations of tissue renin-angiotensin systems in the female reproductive tract. AB - The literature has been provided with conflicting results concerning prorenin in rat plasma. The cause was investigated and we characterized severe methodological problems. Prorenin is in rats measured by its enzymatic property after trypsin activation. Besides activating prorenin, trypsin appeared to degrade renin substrate (angiotensinogen) and formed an unstable 'blank' which interfered in the prorenin measurement. An interfering sulphydryl enzyme was also found in plasma from nephrectomized rats. Identical 'blank' problems were also found in other animals such as pigs and cattle. These problems were solved and a new validated method was developed enabling the study of active renin and prorenin in species other than humans. Plasma prorenin was found primarily to originate from kidneys in rats as in other species although minor contribution of other tissues was likely. Evidence has accumulated during recent years supporting the existence of local tissue RAS's in ovary and uteroplacental unit. The presence of active renin and prorenin in female reproductive organs were verified in cattle and pigs. A marked species difference was found. Secretion of prorenin to plasma only seemed to take place in humans suggesting primarily a local function of reproductive RAS's. The concentration of prorenin varies during follicular maturation and pregnancy in reproductive tissues and fluids. A tissue incubation method was developed to study the bovine ovarian RAS in-vitro. The presence, formation, secretion and degradation of active renin in bovine ovarian follicles in-vitro indicate that active renin--and not prorenin as suggested earlier--is important in mediating effects of tissue RAS. Angiotensin II receptors are also present in high and varying densities in the ovary, uterus and placenta. Several potential effects, mediated through auto- or paracrine actions, are suggested. Ovarian RAS may affect oocyte maturation, ovulation, steroidogenesis and angiogenesis. The uteroplacental RAS may affect decidualization, angiogenesis, hormone synthesis, local blood flow and uterine contraction. Effects may, according to the marked species difference of RAS, vary between species. The understanding of tissue RAS's is in its infancy. The ovary seems to be a very good model in study of tissue RAS's and their possible relations to the bloodborne RAS. Further investigation may also benefit the reproductive endocrinology. PMID- 9408736 TI - Human myocardial and skeletal muscular Na,K-ATPase in relation to digoxin therapy of heart failure. PMID- 9408737 TI - Alphoid repetitive DNA in human chromosomes. AB - The thesis describes the first extensive DNA sequence analysis that demonstrated that the tandemly repeated alphoid DNA in the centromere of the human chromosomes consists of distinct subfamilies and in a number equal to or exceeding the number of chromosomes. The expected presence of only one or a few distinct subfamily on individual chromosomes was supported by the characterization of an extremely well defined subfamily specific for chromosome 7 and represented in the original collection of subfamilies. The pattern of chromosome-specificity breaks down among the acrocentric chromosomes where chromosomes 13 and 21 were found to share one and chromosomes 14 and 22 to share another specific subfamily. By in situ hybridization these subfamilies were shown not to be shared by other chromosomes. The remarkable pairwise pattern of sequence homogenization was present also in the chimpanzee genome raising the question of its biological role. However, the subfamilies on these human and chimpanzee chromosomes are not orthologous but were shown to originate from two evolutionarily different repeat families. It follows that dramatic sequence evolution has occurred in one or both species during or after separation. The sequence evolution might even occur at a higher rate in humans. This possibility was studied in orthologous alphoid sequences on the X chromosome of humans and the great apes. The analysis supports the general view that our closest relative is the chimpanzee and indicates that the rate of recombination is increased in the human repeat DNA. A "molecular clock" running faster in this DNA may have evolutionary implications. Finally, the usefulness of alphoid subfamilies as chromosome-specific markers is illustrated in a cytogenetic dissection of the centromeric region of Robertsonian translocations. The breakpoints were located to satellite III DNA leaving these chromosomes dicentric. The order of the different tandem DNAs on the p-arm of the acrocentric chromosomes could also be established. PMID- 9408738 TI - The Danish Cancer Registry--history, content, quality and use. AB - The Danish Cancer Registry is a population-based registry containing data on the incidence of cancer throughout Denmark since 1943. Reporting of cancer was made mandatory by administrative order in 1987. Details of individual cases of cancer are available according to the 7th revision of the International Classification of Diseases (ICD) for all years, and according to the ICD-O since 1978. A core data set is kept on each individual which includes date of birth, sex, date of cancer diagnosis, method of verification, date of death and cause of death. This paper describes the history of the registry, its data sources and its procedures, including quality control and access to data. Integration of both research activities and registration since the inception of the Registry has maintained the completeness and validity of the data for 1943-1996. PMID- 9408739 TI - The Danish strong analgesic surveillance system. AB - The legal consumption of strong analgesics is increasing and research addressing the complex consequences of this expansion is needed. We describe the Danish strong analgesic surveillance system comprising copies of all prescriptions of strong analgesics purchased at Danish pharmacies. The system is operated by the Public Health officers and has been computerized since 1993. In addition to specific informations about the drugs in question the system comprises personal registration numbers (CPR) of both patients and doctors issuing the prescription. The system therefore allows: 1) a precise and valid identification of the individuals purchasing and prescribing strong analgesics, 2) an option to describe possible changes in consumption over time, and 3) the possibility of a highly reliable record linkage with other Danish population-based data sources. Validation studies are reported and strengths and limitations of the system are discussed. PMID- 9408740 TI - Determinants of general practice utilization in Denmark. AB - The study was undertaken to identify determinants of health services' utilization related to organizational characteristics of the health services, social characteristics of the community, and health and socio-demographic characteristics of the population. We used a follow-up design involving a time dimension of 12 months. A cohort of 3000 50-year old males and females were randomly selected from an urban county near Copenhagen and a rural county in Western Jutland and 65% agreed to participate. Data were collected through: 1) mailed questionnaires to the participants every three months, 2) a mailed questionnaire to all general practitioners in the counties, and 3) the Health Insurance Register. Data were analyzed by multivariate logistic regression analysis. Utilization of health services was found to be strongly associated with gender and health characteristics, especially with respect to functional status and chronic diseases. Health service characteristics, demographic and psycho social factors had very little impact on utilization patterns. The utilization of publicly financed health services was found to be determined by health needs rather than by social, psychological and organizational factors, indicating geographical and social equity in the access to the Danish health services. PMID- 9408741 TI - Elevated levels of sex hormones and sex hormone binding globulin in male patients with insulin dependent diabetes mellitus. Effect of improved blood glucose regulation. AB - In a prospective study we have measured serum levels of sex hormone-binding globulin (SHBG), androgens, oestrogens and gonadotropins in 20 male IDDM patients with the aim of evaluating the effect of improved glycaemic control on these levels and to compare the IDDM patients with an age- and weight-matched healthy non-diabetic control group. The patients were chosen from the male IDDM patients attending the outpatient clinic at the Department of Endocrinology, Odense University Hospital. Glycaemic control was optimized by a trained diabetologist according to the patients' measurements of home blood glucose concentrations and reported hypoglycaemic episodes during a three month period. Prior to regulation the patients, compared to healthy control subjects, had significantly higher serum levels of oestrone (0.29 +/- 0.02 vs 0.16 +/- 0.01 nmol/l (Mean +/- SEM), p < 0.01), 17 beta-oestradiol (0.12 +/- 0.01 vs 0.08 +/- 0.01 nmol/l, p < 0.01), dihydrotestosterone (4.20 +/- 0.18 vs 1.66 +/- 0.09 nmol/l, p < 0.01), total testosterone (20.7 +/- 0.9 vs 17.8 +/- 1.1 nmol/l, p < 0.05) and SHBG (42.3 +/- 2.9 vs 15.5 +/- 3.5 nmol/l, p < 0.05), while the calculated free-testosterone was lower (0.34 +/- 0.02 vs 0.40 +/- 0.02 nmol/l, p = 0.068). After regulation, the patients obtained significantly lower levels of glycosylated haemoglobin (10.4 +/ 0.3 vs 8.9 +/- 0.2%, p < 0.005) and serum fructosamine (1.50 +/- 0.05 vs 1.34 +/ 0.04 nmol/l, p < 0.005) on a higher 24 hour insulin dose (52.7 +/- 4.4 vs 59.2 +/- 4.6 IU/24 h, p < 0.005). Levels of free-testosterone (0.41 +/- 0.04 nmol/l), oestrone (0.33 +/- 0.03 nmol/l), oestradiol (0.14 +/- 0.01 nmol/l), delta 4 androstenedione (4.44 +/- 0.43 vs 3.85 +/- 0.42 nmol/l), and prolactin (249 +/- 24 vs 200 +/- 19 mIU/l) increased compared to values obtained before regulation (all p < 0.05). We conclude that SHBG, testosterone, dihydrotestosterone and oestrogen levels are increased in male IDDM patients. High SHBG levels tend to keep the free fractions of sex hormones within normal limits. During improvement of glycaemic control with insulin, levels of free-testosterone and its bioprecursors and metabolites rise. This may partly be due to the increased daily insulin dose and/or to the improvement in glycaemic control itself. PMID- 9408742 TI - The follicle-stimulating hormone receptor: biochemistry, molecular biology, physiology, and pathophysiology. PMID- 9408743 TI - Insulin resistance and the polycystic ovary syndrome: mechanism and implications for pathogenesis. AB - It is now clear that PCOS is often associated with profound insulin resistance as well as with defects in insulin secretion. These abnormalities, together with obesity, explain the substantially increased prevalence of glucose intolerance in PCOS. Moreover, since PCOS is an extremely common disorder, PCOS-related insulin resistance is an important cause of NIDDM in women (Table 3). The insulin resistance in at least 50% of PCOS women appears to be related to excessive serine phosphorylation of the insulin receptor. A factor extrinsic to the insulin receptor, presumably a serine/threonine kinase, causes this abnormality and is an example of an important new mechanism for human insulin resistance related to factors controlling insulin receptor signaling. Serine phosphorylation appears to modulate the activity of the key regulatory enzyme of androgen biosynthesis, P450c17. It is thus possible that a single defect produces both the insulin resistance and the hyperandrogenism in some PCOS women (Fig. 19). Recent studies strongly suggest that insulin is acting through its own receptor (rather than the IGF-I receptor) in PCOS to augment not only ovarian and adrenal steroidogenesis but also pituitary LH release. Indeed, the defect in insulin action appears to be selective, affecting glucose metabolism but not cell growth. Since PCOS usually has a menarchal age of onset, this makes it a particularly appropriate disorder in which to examine the ontogeny of defects in carbohydrate metabolism and for ascertaining large three-generation kindreds for positional cloning studies to identify NIDDM genes. Although the presence of lipid abnormalities, dysfibrinolysis, and insulin resistance would be predicted to place PCOS women at high risk for cardiovascular disease, appropriate prospective studies are necessary to directly assess this. PMID- 9408744 TI - Insulin-like growth factor-binding proteins in serum and other biological fluids: regulation and functions. PMID- 9408745 TI - Maternal-fetal calcium and bone metabolism during pregnancy, puerperium, and lactation. PMID- 9408746 TI - Analyses of brain tumor cell lines confirm a simple model of relationships among fluorescence in situ hybridization, DNA index, and comparative genomic hybridization. AB - Several techniques are commonly used for genetic analysis of interphase nuclei. Flow cytometry assays the distribution of DNA content in populations of nuclei stained with a DNA-specific fluorochrome. Fluorescence in situ hybridization (FISH) quantifies the number of copies of a specific DNA sequence in single nuclei. Comparative genomic hybridization (CGH) assesses the relative copy number of DNA sequences throughout a test genome by comparing the signal intensities of test and reference DNA samples hybridized to a template of normal metaphase chromosomes. In principle, there are specific relationship among data obtained from these measurements, and combined measurements should provide a more comprehensive view of the sample that is analyzed. We applied these three techniques to nine brain tumor cell lines and find that a model of CGH that includes unsuppressed repeat sequences describes the data well. We estimate that up to 35% of the fluorescence intensity in well-blocked CGH preparations may not represent unique sequences. Taking these factors into account, our results are, in general, mutually consistent, and highlight issues critical for interpreting CGH preparations. PMID- 9408747 TI - Comparative genomic hybridization detects frequent overrepresentation of chromosomal material from 3q26, 8q24, and 20q13 in human ovarian carcinomas. AB - We used comparative genomic hybridization (CGH) to identify recurrent chromosomal imbalances in tumor DNA from 25 malignant ovarian carcinomas and two ovarian tumors of low malignant potential (LMP). Many of the carcinoma specimens displayed numerous imbalances. The most common sites of copy number increases, in order of frequency, were 8q24.1, 20q13.2-qter, 3q26.3-qter, 1q32, 20p, 9p21-pter, and 12p. DNA amplification was identified in 12 carcinomas (48%). The most frequent sites of amplification were 8q24.1-24.2, 3q26.3, and 20q13.2-qter. Other recurrent sites of amplification included 7q36, 17q25, and 19q13.1-13.2. The most frequent sites of copy number decreases were 5q21, 9q, 17p, 17q12-21, 4q26-31, 16q, and 22q. Underrepresentation of 17p was observed in six of 16 stage III/IV tumors, but in none of seven stage I/II tumors, suggesting that this change may be a late event associated with the transition of ovarian carcinomas to a more metastatic disease. Overrepresentation of 3q26.3-qter, 5p14-pter, 8q24.1, 9p21 pter, 20p, and 20q13.2-qter and underrepresentation of 4q26-31 and 17q12-21 also tended to be more common in advanced-stage tumors. All ten grade 3 tumors had copy number increases involving 8q24.1, compared to only three of nine grade 2 tumors. Overrepresentation of 3q26.3-qter and 20q13.2-qter was also observed at a higher frequency in high-grade tumors. One of the two LMP tumors displayed chromosomal alterations, which consisted of overrepresentation of 5p and 9p only. Taken collectively, these findings and data from other CGH studies of ovarian cancers define a set of small chromosome segments that are consistently over- or underrepresented and, thus, highlight sites of putative oncogenes and tumor suppressor genes that contribute to the pathogenesis of these highly malignant neoplasms. PMID- 9408748 TI - Human/mouse microcell hybrid based elimination test reduces the putative tumor suppressor region at 3p21.3 to 1.6 cM. AB - We have previously identified an approximately 7 cM long common eliminated region (CER), involving the 3p21.3 markers AP20R, D3S966, D3S3559, D3S1029, WI-7947, D3S2354, AFMb362wb9, and D3S32, in human chromosome 3/A9 mouse fibrosarcoma microcell hybrid (MCH) derived SCID mouse tumors. We now report the results of our more detailed analysis on 24 SCID mouse tumors derived from two MCH lines that originally carried intact human chromosomes 3. They were analyzed by fluorescence in situ hybridization (FISH) painting and PCR, using 24 markers covering the region between D3S1611 and D3S13235 at 3p22-p21.2. D3S32 and D3S2354 were regularly eliminated during in vivo tumor growth, whereas the other 22 markers, D3S1611, ACAA, D3S1260, WI-692, AP20R, D3S3521, D3S966, D3S1029, D3S643, WI-2420, MSTI. GNAI2, D3S1235, D3S1298, GLBI, WI-4193, D3S3658, D3S3559, D3S3678, WI-6400, WI-7947, and WI-10865, were regularly retained. We have defined a common eliminated region of approximately 1.6 cM (designated as CER1) inside the 7 cM CER described earlier. CER1 is flanked distally by D3S1029 and proximally by D3S643. PMID- 9408749 TI - Characterization of a hairy cell leukemia-associated 5q13.3 inversion breakpoint. AB - Previous cytogenetic analysis has indicated that chromosome anomalies involving the 5q13 band are common in hairy cell leukemia (HCL), occurring in approximately 1/3 of the patients. The data suggest that 5q13.3 is likely to harbor a gene involved in the transformational event of this disease. We selected a constitutional inv(5)(p13.1q13.3) in a patient with HCL as the starting point in an attempt to identify the relevant gene in 5q13.3. By using double color interphase fluorescence in situ hybridization (FISH) techniques, we have identified two cosmid probes from a chromosome 5-specific library that flank the 5q13.3 inversion breakpoint proximally and distally. Pulsed field gel electrophoresis (PFGE) and interphase FISH experiments suggest that the two markers are at a distance of no more than 300 kb. YAC probes covering a 21 Mb region at 5q13 were used to map the 5q13.3 inversion breakpoint and the breakpoint is located within the D5S646-D5S620 region. Two non-chimeric YACs have been identified that span the breakpoint. FISH analysis revealed that four other patients with cytogenetic aberrations of 5q carried inversions/deletions that involved the same 5q13.3 breakpoint region. The identification of a gene involved in hairy cell leukemogenesis in this region will be of major importance in the elucidation of the transformational events of HCL. PMID- 9408750 TI - Deletion mapping defines three discrete areas of allelic imbalance on chromosome arm 8p in oral and oropharyngeal squamous cell carcinomas. AB - Deletions on chromosome arm 8p, as defined by allelic imbalance, are a frequent event in many different types of malignant tumors, including those of the head and neck. These regions are thought to harbor tumor suppressor genes. In order to define a high-density deletion map of this chromosomal arm in oral and oropharyngeal squamous cell carcinomas, we have tested for allelic imbalance in 35 such tumors with 22 short tandem-repeat polymorphisms. Overall, 21 (60%) of the 35 tumors showed allelic imbalance at one or more loci on chromosome arm 8p. Interstitial deletions defined three discrete areas of deletion: at 8p23, 8p22, and 8p12-p21. Tumors of TNM stages II-IV showed a significantly higher frequency of allelic imbalance on 8p than did TNM stage I tumors. Our data suggest that there are least three tumor suppressor loci on chromosome arm 8p that may be implicated in oral carcinogenesis. Furthermore, inactivation of such genes may be associated with high-grade tumors. PMID- 9408751 TI - Four regions of allelic imbalance on 17q12-qter associated with high-grade breast tumors. AB - Rearrangements or loss of chromosome 17 are frequent events in breast tumors. Chromosome 17 contains at least four genes implicated in breast cancer (TP53, ERBB2 (Her2/neu), BRCA1, and NM23), as well as other putative tumor suppressor genes and oncogenes implicated in loss of heterozygosity or allelic imbalance studies. Allelic imbalance represents the addition or loss of genetic material in tumor samples, providing circumstantial evidence for the location of cancer related genes. We have analyzed a panel of 85 breast tumor/normal tissue pairs with 21 PCR-based short tandem repeat (STR) markers located at 17q12-qter to more precisely define regions of allelic imbalance and to determine their relation to clinical parameters. Our analysis revealed at least four common regions of allelic imbalance: proximal to BRCA1, including D17S800 (17q12); distal to NM23 around D17S787 (17q22); near the growth hormone (GH) locus, at D17S948 (17q23 24); and between markers D17S937 and D17S802 (17q25). These data also reveal that loss (or gain) of 17q genetic material correlates with poorly differentiated (grade III) tumors (P = < 0.001), high S phase fraction (P = 0.034), and positive TP53 immunohistochemical staining (P = 0.011). However steroid receptor status, ERBB2 (Her2/neu) staining, and aneuploidy do not correlate with allelic imbalance at 17q. PMID- 9408752 TI - Molecular analysis of the t(8;14)(q24;q11) chromosomal breakpoint junctions in the T-cell leukemia line MOLT-16. AB - The MOLT-16 cell line was established from the leukemic cells of a patient with T cell acute lymphoblastic leukemia and contains a t(8;14)(q24;q11) resulting in juxtaposition of sequences downstream of the MYC gene on chromosome 8 and the J region of the T-cell receptor alpha chain gene (TCRA) on chromosome 14. The reciprocal translocation involved a complex rearrangement with two chromosome breakpoints within the TCRAJ region on chromosome 14, resulting in inversion of a 1.4 kb DNA fragment between the two breakpoints. The 5' border of the inversion joints with another segment of chromosome 14, whereas the 3' border joins with a region of chromosome 8 located at least 257 kb downstream of MYC. Extensive deletions have occurred on both chromosomes 8 and 14 in conjunction with the translocation. To investigate the possible involvement of the V(D)J recombinase in this translocation, we analyzed the nucleotide sequences surrounding the translocation breakpoints. The breakpoint on chromosome 14 occurs between a segment coding for a TCRAJ sequence and its hepatamer-nonamer signal. Heptamer nonamer consensus sequences are also identified on chromosome 8 adjacent to the breakpoint. Inserted N and P nucleotides are observed at the breakpoint junctions. PMID- 9408753 TI - Increased copy number at 17q22-q24 by CGH in breast cancer is due to high-level amplification of two separate regions. AB - Studies by comparative genomic hybridization (CGH) have defined a chromosomal site at 17q22-q24 that is often overrepresented in breast cancer, neuroblastoma, and several other tumor types. Due to the limited resolution and dynamic range of CGH, it remain unclear whether this gain reflects high-level amplification of small subregion(s) or low-level gain of most of the distal 17q. We used 32 physically mapped 17q probes to construct more accurate copy number profiles for 14 breast cancer cell lines by interphase fluorescence in situ hybridization (FISH). Six cell lines (43%) showed an increased copy number of the 17q-22q24 region by CGH, and seven (50%) by FISH. FISH copy number profiles had a substantially higher dynamic range than did CGH profiles. FISH revealed two independent, highly amplified regions (A and B) at 17q23, separated by about 5 Mb of non-amplified DNA. These regions were distinctly telomeric from the ERBB2 gene locus. However, region A was often co-amplified with ERBB2, whereas B was amplified in cell lines that showed no ERBB2 amplification. We conclude that distal 17q gains recently discovered in breast cancer by CGH are due to high level amplifications of two different regions at 17q23. This chromosomal region has previously been reported to undergo allelic loss and therefore was thought to harbor a tumor suppressor gene. The present FISH data provide support for the presence, and a starting point for the positional isolation, of 17q23 genes whose upregulation by amplification may play a role in the progression of breast cancer and many other tumor types. PMID- 9408755 TI - Comparative genomic hybridization reveals frequent gains of 20q, 8q, 11q, 12p, and 17q, and losses of 18q, 9p, and 15q in pancreatic cancer. AB - Comparative genomic hybridization (CGH) was used to screen for genomic imbalances in 24 exocrine pancreatic carcinomas, including II low-passage cell lines (4-8 subcultures) and 13 uncultured samples. Aberrations were found in all cell lines and in seven of the 13 biopsies. The most frequent changes in the cell lines were gains of 20q (91%), 11q (64%), 17q (64%), 19q (64%), 8q, 12p, 14q, and 20p (55%), and losses of 18q (100%), 9p (91%), 15q(73%), 21q (64%), 3p (55%), and 13q (55%). High-levels gains (tumor to normal ratio over 1.5) were detected at 3q, 6p, 7q, 8q, 12p, 19q, and 20q. Among the tumor biopsies, overrepresentations of 7p and 8q were most common (31%), followed by 5p, 5q, 11p, 11q, 12p, and 18q (23%), whereas the most frequent losses involved 18p and 18q (31%) and 6q and 17p (23%). The genetic changes in nine samples obtained from metastatic lesions did not differ significantly from those in 15 primary carcinomas. Most of the gains and losses detected in this CGH study correspond well to those identified in previous cytogenetic and molecular genetic investigations of pancreatic carcinomas. However, frequent gain of 12p and loss of 15q have not been previously reported. Molecular genetic analyses of these chromosome arms are warranted, and may lead to the discovery of novel genes important in pancreatic carcinogenesis. PMID- 9408754 TI - Allelic loss at BRCA1, BRCA2, and adjacent loci in relation to TP53 abnormality in breast cancer. AB - Cells with abnormal TP53 lose cell cycle checkpoints, resulting in genomic instability and neoplastic transformation. However, the evidence linking the tumor-specific targets of genomic alteration to an abnormal TP53 is limited. The present study tested the hypothesis that TP53 abnormalities are correlated with an increased frequency of deletion of breast cancer susceptibility loci (17q and 13q) in breast carcinomas. Tumors from 90 patients were examined for TP53 abnormality and loss of heterozygosity (LOH) at 11 loci on 17q (17q11.2-21) and 13q (13q12-14), including the loci for BRCA1 and BRCA2. A higher frequency of LOH was consistently found at 17q or 13q loci in tumors with an abnormal TP53. The increased LOH in relation to TP53 abnormality was statistically significant at the BRCA1, D17S588, and D13S267 loci (P < 0.05) but not at the locus for BRCA2 (P = 0.64). These observations imply a possible link between an abnormal TP53 and specific genomic deletions of breast cancer susceptibility loci, which may provide clues to the role of TP53 during breast tumorigenesis. PMID- 9408756 TI - Hyperdiploidy and E2A-PBX1 fusion in an adult with t(1;19)+ acute lymphoblastic leukemia: case report and review of the literature. AB - The t(1;19)(q23;p13), detected cytogenetically in 5-6% of cases, is one of the most common translocations in childhood acute lymphoblastic leukemia (ALL). Most t(1;19)+ ALLs are pseudodiploid or contain fewer than 50 chromosomes, are classified as pre-B based on expression of cytoplasmic, but not surface, immunoglobulin (clg+/slg-), express a characteristic pattern of cell surface antigens, and contain E2A-PBX1 fusion mRNAs. A minority of cases are early pre-B (clg-/slg-), do not express the characteristic pattern of cell surface antigens, and lack E2A-PBX1 fusion mRNAs. These latter cases are frequently hyperdiploid, with a modal chromosome number of 55-57. The incidence of the t(1;19) in adults with ALL (approximately 3%) appears to be similar to that observed in children, but the genetic and immunophenotypic features of adult t(1;19)+ ALL have not been described extensively. We report a case of t(1;19)+ ALL occurring in a 38-year old man in the setting of hyperdiploidy > 50. Despite this feature, this case was pre-B, conformed to the classic t(1;19) immunophenotype, and expressed E2A-PBX1 fusion mRNAs. This prompted us to review the published literature on ploidy and genetic features of t(1;19)+ ALLs. Overall, E2A-PBX1 fusion occurred in 95% (102/107) of t(1;19)+ B-lineage ALLs with 50 or fewer chromosomes, 80% of which were pseudodiploid, vs. only 25% (2/8) of t(1;19)+ ALLs with more than 50 chromosomes. PMID- 9408757 TI - Matrix-based comparative genomic hybridization: biochips to screen for genomic imbalances. AB - Comparative genomic hybridization (CGH) to metaphase chromosomes has been widely used for the genome-wide screening of genomic imbalances in tumor cells. Substitution of the chromosome targets by a matrix consisting of an ordered set of defined nucleic acid target sequences would greatly enhance the resolution and simplify the analysis procedure, both of which are prerequisites for a broad application of CGH as a diagnostic tool. However, hybridization of whole genomic human DNA to immobilized single-copy DNA fragments with complexities below the megabase pair level has been hampered by the low probability of specific binding because of the high probe complexity. We developed a protocol that allows CGH to chips consisting of glass slides with immobilized target DNAs arrayed in small spots. High-copy-number amplifications contained in tumor cells were rapidly scored by use of target DNAs as small as a cosmid. Low-copy-number gains and losses were identified reliably by their ratios by use of chromosome-specific DNA libraries or genomic fragments as small as 75 kb cloned in PI or PAC vectors as targets, thus greatly improving the resolution achievable by chromosomal CGH. The ratios obtained for the same chromosomal imbalance by matrix CGH and by chromosomal CGH corresponded very well. The new matrix CGH protocol provides a basis for the development of automated diagnostic procedures with biochips designed to meet clinical needs. PMID- 9408758 TI - Characterization of a t(10;12)(q24;p13) in a case of CML in transformation. AB - We have used Southern blotting and fluorescence in situ hybridization (FISH) to define the breakpoints of a reciprocal translocation, t(10;12)(q24;p13), acquired as a secondary abnormality in a patient with Philadelphia chromosome positive chronic myeloid leukemia (CML) in transformation. A YAC clone that spanned the breakpoint at 12p13 was identified; this YAC included the CDKN1B gene but did not include ETV6. Neither ETV6 nor CDKN1B was rearranged, as determined by FISH and Southern blotting; however, a small deletion encompassing the translocated CDKN1B allele was detected. Analysis of two candidate genes at 10q24, HOX11 and NFKB2 suggested that they are not involved in this translocation. The preliminary mapping of breakpoints in this case demonstrated that they are different from an apparently identical translocation identified previously in a patient with myelodysplastic syndrome. The identification of the split YAC and small deletion should enable a more focused search for a gene or genes that may contribute to progression from chronic phase to blast crisis in CML. PMID- 9408759 TI - Detection of chromosomal aberrations in seminomatous germ cell tumours using comparative genomic hybridization. AB - Comparative genomic hybridization (CGH) was used to evaluate tissue specimens from 16 seminomas in order to elucidate the pathogenesis of germ cell tumours in males. A characteristic pattern of losses and gains within the entire genomes was detected in 94% of the seminomas by comparing the ratio profiles of the tumours with a standard of cytogenetically normal genomic DNA. Losses represented 43% of the total number of alterations often affecting chromosomes and chromosome arms 4, 5, 11, 13q, and 18q. Gains amounted to 57% and were often observed on 1q, 7, 8, 12, 14q, 15q, 21q, and 22q. Aberrations of 12p and 21q appeared most consistently. Results from CGH analysis displayed no relationship to the clinical stages of the malignancy. Some rare aberrations appeared, however, only in clinical stage II and in tumours showing relapse in the contralateral testis following orchiectomy, although the alterations were not present in all of the tumours in question. Losses of 16q13-21 and gains of 9q22.1-22.2 were demonstrated in both groups, while loss of 16p12 and gains of 6p21 and 6q23.3-24 were detected in the latter group as well. In conclusion, a specific pattern of chromosomal alterations was demonstrated in the seminomas by improved detection criteria, which increased specificity and sensitivity. The rare aberrations, which appeared only in tumours in improved detection criteria, which increased specificity and sensitivity. The rare aberrations, which appeared only in tumours in clinical stage II and relapsed tumours, may be linked to tumour progression, invasiveness, and bilateral disease. PMID- 9408760 TI - 1p and 3p deletions in meningiomas without detectable aberrations of chromosome 22 identified by comparative genomic hybridization. AB - Meningioma is a common tumor of the meninges covering the central nervous system. Although generally a benign tumor, meningioma often recurs and is malignant in 5 10% of all cases. Loss of chromosome 22 loci, and specifically inactivation of the NF2 tumor suppressor gene, is considered one of several critical steps in the tumorigenesis of meningioma. However, cytogenetic and molecular investigations have failed to detect either aberrations of chromosome 22 or mutations in the NF2 gene in approximately 40% of all tumors, thus making it apparent that an alternative mechanism(s) is responsible for the development of a large fraction of meningiomas. This subset of meningiomas is not distinct with regard to clinical and histopathological features from tumors showing deletions on chromosome 22. It is, therefore, important to attempt the elucidation of molecular pathway(s) that may operate in the tumorigenesis of these tumors. We used comparative genomic hybridization (CGH) to identify regions of the genome other than chromosome 22, contributing to the development of meningioma. We analyzed 25 tumors that had undergone detailed LOH analysis on chromosome 22 and were shown to contain no detectable deletions. Two benign, malignancy grade I, meningiomas showed concurrent deletion of 1p and 3p. These results suggest that loss of both 1p and 3p may contribute to meningioma tumorigenesis. This may represent genetic changes that are alternative to deletions on chromosome 22. PMID- 9408761 TI - Der(16)t(1;16)(q21;q13) as a secondary structural aberration in yet a third sarcoma, extraskeletal myxoid chondrosarcoma. AB - Cytogenetic analysis of an extraskeletal myxoid chondrosarcoma revealed a der(16)t(1;16)(q21;q13) in addition to the t(9;22)(q22;12) described as characteristic for this chondrosarcoma clinicohistopathologic subtype. An identical der(16) has been identified as the most common secondary structural aberration in Ewing's sarcoma and alveolar rhabdomyosarcoma. PMID- 9408762 TI - Splenectomy for haematological diseases--a single institution experience. AB - OBJECTIVES: To evaluate our experience with splenectomy for haematological disease over 15 years, focusing on the diseases and indications requiring surgery, the complications and the haematological results. DESIGN: A single institution retrospective analysis. SETTING: Departments of Surgery, Internal Medicine and Haematology, Rabin Medical Center, Hasharon Hospital, Petah-Tikva, Israel. PATIENTS: Sixty-nine patients undergoing splenectomy for haematological diseases between 1980 and 1994. PARAMETERS STUDIED: (1) the patient characteristics; (2) the haematological disorders and the indications for splenectomy; (3) the splenic size; (4) perioperative complications; (5) the haematological and other results of surgery. RESULTS: The common disease requiring splenectomy in our patient population was immune thrombocytopenic purpura (ITP). Eighteen patients (26%) suffered from ITP, 12 (17.4%) had lymphoproliferative (LP) disorders, including 11 patients with non-Hodgkin's lymphoma (NHL), 9 (13%) immune haemolytic anaemia (IHA), 8 (11.6%) Hodgkin's disease (HD), and 8 patients with myeloproliferative diseases (MPD). Fifty-two patients (75.4%) underwent surgery for therapeutic purposes while 25 patients (36.2%) underwent diagnostic surgery. Eight patients were operated on for both indications. Patients with HD and ITP were younger (mean age in the 30th year) as opposed to patients with congestive splenomegaly and LP (mean age in the 60th year). Most patients with ITP, NHL and IHA were female. Patients with HD and ITP had a small spleen (< 300 g), in contrast with HCL and MPD (> 2400 g). No perioperative mortality was observed. Twenty-one patients (30.4%) experienced perioperative complications, including bleeding in 9 patients (13%) and infection in 10 patients (14.5%). Of the 34 patients evaluable for platelet response to splenectomy, 26 (76.5%) achieved complete response (CR), including 15 of 18 patients with ITP. Eleven of the 17 (64.7%) anaemic patients evaluable for haemoglobin (Hb) response achieved CR. Eight of the 9 leucopenic patients obtained CR. In 15 patients, splenectomy established the diagnosis, including 8 patients with NHL. In 2 others, surgery confirmed the tentative diagnosis. In 2 of the 8 HD patients the spleen was found to be involved. CONCLUSIONS: From both therapeutic and diagnostic standpoints splenectomy is a relatively safe and effective procedure. However, no long-term follow-up data are provided to allow conclusions regarding the long-term prognosis of diseases such as myelo- or lymphoproliferative disorders. PMID- 9408763 TI - Refractory anaemia with ringed sideroblasts concurrent with multiple myeloma--a brief review of the recent literature. AB - The report describes a patient in whom myelodysplastic syndrome and multiple myeloma (MM) were simultaneously present. This patient manifested an IgA-lambda type of MM concurrent with a refractory anaemia with ringed sideroblasts (RARS) without prior therapy. His bicytopenia could not be improved by vitamin B6 regardless of a reduced serum vitamin B6 concentration. A review of the literature suggests that myelodysplastic syndrome (MDS), chronic neutrophilic leukaemia (CNL) and idiopathic myelofibrosis (IMF) are the most frequent disorders associated with MM. The IgA type seems to be associated more commonly with these disorders. The mechanisms responsible for the development of plasma cell proliferation are diverse; the neoplastic transformation of a common progenitor, the involvement of the lymphoplasmacytic system and/or chronic reticuloendothelial stimulation may play a role in the occurrence of such hybrid haematological disorders. PMID- 9408765 TI - Congenital systemic Langerhans cell histiocytosis (report of two cases). AB - Two cases of congenital systemic Langerhans cell histiocytosis (LCH), diagnosed and treated in our department from June 1995 until May 1996, are described. The cases concern two neonates (one female and one male) born with necrotic lesions and skin nodules. The diagnosis was confirmed by skin biopsy which showed diffuse infiltration by CD1 antigen and S-100 protein positive histiocytes. The babies didn't present with anemia, hepatosplenomegaly or lymphadenopathy. Hepatic and renal function were normal. In both infants skeletal survey showed no lytic lesions but chest X-rays and high resolution computerized tomography (HRCT) scan revealed diffuse mottling of both lung fields. Bone marrow aspiration showed the presence of histiocytes in percentages of 6% and 10%, respectively. Both babies were treated with prednisolone 1 mg/kg body weight for three months. The first child who is 20 months old, is now well with resolution of skin and pulmonary lesions occurring within one month of the initiation of steroids, while the second, who presented spectacular resolution of skin lesions within the first three weeks of therapy, is also in excellent condition five months after completion of treatment. We conclude that congenital LCH has to be suspected in neonates with persisting skin lesions. If the disease is systemic but without organ dysfunction, treatment with steroids may be beneficial. PMID- 9408764 TI - Administration of MINE protocol in untreated patients with intermediate and high grade non-Hodgkin's lymphoma. AB - In recent years, more effective and less toxic treatment protocols have been developed to increase the cure rates in intermediate and high grade non-Hodgkin's lymphoma (NHL). This study was undertaken to investigate the efficacy and toxicity of MINE (ifosfamide, mesna, mitoxantrone and etoposide) combination chemotherapy in patients with intermediate and high grade NHL. Twenty-one patients (16 male, 5 female; age between 26 and 70 years) with NHL were included in the study. An overall response rate of 73% and complete response rate of 56% were achieved and survival rate for responding patients was 80% at the 48th month. Side effects including mild myelosuppression, nausea/vomiting and alopecia were observed. MINE combination seems to be effective and well tolerated without significant toxicity as a first-line therapy in patients with intermediate or high grade NHL. PMID- 9408766 TI - 5'-nucleotidase and adenosine deaminase activities in blood of patients with unstable angina pectoris. AB - During ischaemia and hypoxia adenosine is released from cardiac cells. Adenosine is the end product of 5'-nucleotidase activity. We were interested in how this enzyme activity in plasma of patients with unstable angina pectoris, causes short term ischaemia. 5'-Nucleotidase activity in plasma was determined using a standard diagnostic kit from Sigma. Furthermore, we studied the activity of adenosine deaminase in plasma, granulocytes, lymphocytes and erythrocytes by the methods of Hopkinson [1]. It was found that 5'-nucleotidase activity was increased by about 43% in plasma. The activity of adenosine deaminase (ADA) in plasma increased by 6%, but in granulocytes, lymphocytes and erythrocytes decreased by about 24, 19 and 10.6%, respectively. We concluded that a large increase in 5'-nucleotidase activity may be caused by activation of 5' ectonucleotidase in blood cells by ischaemia. However, the decrease in ADA activity in blood cells may be associate with the adenosine metabolism. PMID- 9408767 TI - Investigation of malondialdehyde formation and antioxidant enzyme activity in stored blood. AB - In the present study, fresh blood obtained from six healthy adult male donors was investigated for erythrocyte malondialdehyde (MDA) formation and antioxidative enzyme activity. Plasma and erythrocyte membrane lipid peroxidation were measured by MDA formation. Erythrocyte superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) levels were determined in whole blood stored for transfusion purposes. Erythrocyte and plasma MDA levels increased significantly during the storage period from day 3 to day 19 and, after that, stayed unchanged. Erythrocyte MDA increased 100% on day 7 when compared to day 1. The GSH-Px activity significantly decreased after day 9 and SOD decreased after day 13. The reduction in enzyme activities continued until day 27. Our results showed significant alteration in erythrocyte membrane and plasma MDA formation and intracellular antioxidant enzyme status in whole blood used for transfusion. However, we do not know whether such alterations have clinical importances for the recipient. PMID- 9408768 TI - A simple morphological score for the quality control of platelet concentrates. AB - The percentage of platelets that retain discoid form correlate with their post transfusion viability and therefore they may act as a simple indicator for the quality of the stored platelet concentrates (PCs). The morphological score (MS) of platelets, given by Kunicki, requires the index of different forms of platelets: discoid, spherical, starlike and bizarre. In the present study, we intended to clarify whether the morphological score of discoid platelets alone could serve as a method for the estimation of PCs quality. During the first five days after preparation, samples of standard platelet concentrates (SPCs) and of leucodepleted platelet concentrates (LDPCs) were analyzed morphologically by immersion light microscopy using smears stained by the May-Grunwald-Giemsa method. In addition to the MS, an original Modified Morphological Score (MMS), for a population of 200 platelets, was used to count platelets with the discoid shape. The mean MMS ranged (first to fifth day) from 192.6 to 166.6 with SPCs and from 197.2 to 185.4 with LDPCs. There is a significant difference (ANOVA) between: (a) PCs prepared by different techniques (p < 0.01); (b) the quality of the same PCs during the storage (p < 0.0001). PMID- 9408769 TI - Dual image of myosin and actin in human neutrophils during movement by fluorescence microscopy. AB - It is well known that myosin and actin play important roles as a motile apparatus in cells, and as a cytoskeleton of cell structure in non-muscle cells. The purpose of this study was simultaneously to demonstrate the intracellular dynamics of myosin and actin in human neutrophils during movement. We designed a double-fluorescence labelling procedure utilising fluorescein isothiocyanate and rhodamine. Myosin was labelled with the green-fluorescence, F-actin was labelled with the red-fluorescence and the coexisting myosin and F-actin were labelled with yellow-fluorescence. We obtained the dual image of myosin and F-actin in human neutrophils during movement by this procedure through a conventional fluorescence microscope. Dual image can interpret the relationship between myosin and actin in human neutrophils during movement, however, further studies are required to elucidate the contractile mechanism in such cells. PMID- 9408770 TI - Reactivity of a transient autoantibody inhibited by ionized calcium. AB - A 53-year-old female suffering from severe anaemia following falciparum malaria required blood transfusion. Her blood specimen in EDTA showed strong autoagglutination giving problems in the preliminary grouping by a slide test. She was grouped, however, as B Rh-D positive without difficulty by the cell and serum groupings done using clotted blood. Two units of B Rh-D positive blood compatible with the patient's serum were transfused without untoward reaction. The patient's blood collected a fresh in different anticoagulants showed autoagglutination in the plasma with EDTA or citrate, but not in heparin collection or in serum, indicating a possible role of ionized calcium as an inhibitor of reactivity. Addition of 0.02 M calcium chloride solution to the plasma remarkably weakened the strength of agglutination, evidently supporting this hypothesis. Autoantibody, identified as anti-Pra, was transient in nature and possibly associated with the malarial infection, as neither the antibody nor the malarial parasites were detected in her blood in follow up studies four months later. PMID- 9408771 TI - Remission of hairy cell leukemia without treatment. AB - A patient is described with hairy cell leukemia, in whom a remission occurred spontaneously or in association with Legionnaires or subsequent HIV infection. Possible pathophysiological mechanisms are reviewed. Currently it is seventeen years since the diagnosis of hairy cell leukemia and fourteen years since the infusion of HIV infected blood. Hairy cell leukemia has yet to return and HIV infection has not progressed to AIDS. PMID- 9408772 TI - Successful outcome of aspergillus brain abscess in a patient who underwent bone marrow transplantation for aplastic anemia. AB - We report the course of an aspergillus brain abscess in an 18-year-old female patient who underwent bone marrow transplantation for aplastic anemia. The abscess was discovered on day 35 post-transplant, in a cranial computerized tomography (CT) scan performed for the evaluation of an unexplained headache. Meanwhile, she was receiving broad-spectrum antibacterials and liposomal amphotericin B for a right upper pulmonary lobe infiltrate. A percutaneous puncture of the cerebral lesion was performed; fungal elements were seen in the pus obtained and its culture yielded A. fumigatus. The dose of amphotericin B was increased, intraconazole was added and two more punctures were done. With these antifungals, the abscess regressed significantly; so, amphotericin B was discontinued after a cumulative dose of 6775 mg but intraconazole was maintained at 400 mg/day. At the last follow-up, seventeen months after detection of the abscess, the patient was well, without symptoms and the cerebral lesion diminished to a very small, thick-walled CT image. PMID- 9408773 TI - Assisted suicide, community, and the common good. PMID- 9408774 TI - Making the case for culturally appropriate community services: Puerto Rican elders and their caregivers. AB - This study of the caregiving arrangements of 214 disabled elderly people in a large northeastern city found that the recipients of care were severely disabled, particularly in instrumental activities of daily living, yet had limited sources of informal care and used formal services minimally. The authors discuss the need to use a structural adaptation approach and to develop social services that are culturally sensitive and acceptable to both elders and their caregivers. PMID- 9408775 TI - The hospital social work self-efficacy scale: a partial replication and extension. AB - The Hospital Social Work Self-Efficacy Scale (HSWSE) was developed to assess social workers' confidence about their ability to perform hospital social work. The scale was originally tested with master's-level social work students. This partial replication and extension included both students and professional social workers. In addition, we used a new scale to assess construct validity and added another hospital to assess external validity. In general, findings continue to support the use of the HSWSE. In addition, anecdotal evidence suggests that this is an outcome measure that practitioners see as easy to use, specific, and highly relevant. PMID- 9408776 TI - Self-esteem instability and its implications for HIV prevention among gay men. AB - This study, using a modified State Self-Esteem Scale (SSES), examined self-esteem instability and its association with risky sexual behavior among 455 gay and bisexual men. A self-administered questionnaire included the SSES and measures of intimacy, loneliness, social support, and ways of coping. Analysis of variance showed that the self-esteem of participants who recently engaged in unprotected anal intercourse with nonprimary partners was more unstable than the self-esteem of participants who did not engage in unprotected anal intercourse. Self-esteem instability was associated with higher avoidance coping, higher loneliness, and lower social support. Episodes of self-esteem injury might motivate some gay men to engage in risky sex. HIV prevention strategies with gay men should target the quality of their interpersonal relationships and community supports. PMID- 9408777 TI - Barriers to school-based health care programs. AB - Although school-based health care programs (SBHCPs) provide affordable and accessible health care to children and adolescents and are known to improve school attendance, a variety of barriers affect their development. Focus groups were conducted in three schools in Louisiana to demonstrate how barriers can affect the initiation and development of SBHCPs. Each school-based program was in a different stage of development. Identifying potential barriers and developing strategies to overcome them can enhance already existing SBHCPs and make it easier for new programs to begin. The social worker serves as an important ally in the development of SBHCPs and is a necessary part of the school-based health care team. PMID- 9408778 TI - The SAFE project: community-driven partnerships in health, mental health, and education to prevent early school failure. AB - This article presents a case study of an innovative school-based health and mental health project that prevents early school failure in one county in Oklahoma. Success is attributed to social work development of broad-based partnerships involving families, schools, communities, and public policy officials. Citizen-driven, these partnerships have meshed previously fixed institutional boundaries in health, mental health, and education to prevent early school failure. The article describes school-family partnerships that form the core of the project's service intervention model. Statistics on service activities and outcomes are presented, along with a discussion of lessons learned for implementation of the project. PMID- 9408779 TI - When people with pre-existing disabilities age in place: implications for social work practice. AB - We focus on a population of people with disabilities who are "aging in place," that is, individuals aging with pre-existing physical disabilities. We distinguish between those who experience prolonged aging and others who experience accelerated aging. A brief overview of people aging with disabilities and selected background information on the increasing linkages between the aging and disability communities is provided. Four case examples illustrate the practice implications faced by social workers in partnering with people with pre existing disabilities and in being sensitive to their desires concerning aging in place. PMID- 9408780 TI - A young women's support group: prevention of a different kind. PMID- 9408781 TI - The Medicaid sweater, children's health, and the tiny hole. PMID- 9408782 TI - It isn't funny! PMID- 9408783 TI - Developing effective helping relationships in health education practice. AB - Health educators who interact directly with the people they serve must be able to establish effective relationships. Helping relationships are effective if they facilitate clients' progress toward health-promoting goals. Health educators are usually well versed in learning activities and spend a significant proportion of their time interacting with clients. However, many health educators have never received explicit training in how to establish effective formal helping relationships. Research on social influence processes has provided a set of empirical findings that suggest interpersonal behaviors that are likely to maximize the effectiveness of formal helping relationships. This literature indicates that formal helping relationships characterized by interpersonal behaviors that enhance client self-esteem and feelings of control are most effective in helping clients achieve specific goals. Interestingly, enhancement of self-esteem and feelings of control are consistent with many definitions of personal empowerment. Since the social influence and empowerment literatures come from very different intellectual roots and have different approaches to power and influence, their convergence is especially notable. These literatures combine to establish the bases for proposing two essential components of effective helping relationships: (1) providing unconditional acceptance and positive regard for clients, and (2) sharing power and control through participatory processes. PMID- 9408784 TI - Effective helping relationships: the role of power and control. PMID- 9408785 TI - A simple reinforcement strategy for increasing attendance at a fitness facility. AB - The purpose of the present study was to examine the effect of a previously untested reinforcement strategy (i.e., 1 month's free membership) on attendance at a fitness facility. Participants were paying members of a fitness facility randomly assigned to control (n = 100), placebo (n = 100), and reinforcement (n = 100) conditions. The control condition received no intervention; the placebo condition received a letter by mail; and the reinforcement condition received the same letter by mail, except that it included an additional paragraph instructing them that they could earn 1 month's free membership if they attended the fitness facility at least 12 times in the next month. Attendance was monitored for 1 month baseline and postintervention by using the facility's computer system. Analysis of variance, t tests, and chi-square analysis all revealed that the reinforcement condition had the best attendance record during the intervention period. These preliminary results shed some light on increasing attendance at a fitness facility. Discussion highlighted the practical implications of the findings and offered directions for future research. PMID- 9408786 TI - The Child and Adolescent Trial for Cardiovascular Health (CATCH): intervention, implementation, and feasibility for elementary schools in the United States. AB - The Child and Adolescent Trial for Cardiovascular Health (CATCH) was the largest school-based field trial ever sponsored by the National Institutes of Health. The trial demonstrated positive changes in the school food service and physical education program, as well as in students' cardiovascular health behaviors. Because the CATCH intervention programs were implemented in 56 schools (in four states) that were typical of schools throughout the United States, their reception by schools and degree of implementation provide evidence about their feasibility for schools nationally. Extensive process evaluation data were collected from students, teachers, school food service personnel, and physical education specialists throughout the three school years of the CATCH intervention. Four of the CATCH programs--school food service, physical education, classroom curricula, and home programs--were assessed over the three school years. The process data provide information on participation, dose, fidelity, and compatibility of the CATCH programs in the intervention schools for these programs. High levels of participation, dose, fidelity, and compatibility were observed for the four programs during the 3 school years. CATCH emerges as a model of a feasible multilevel health promotion program to improve eating and exercise behaviors for elementary schools in the United States. PMID- 9408787 TI - HIV testing among Haitian women: lessons in the recognition of risk. AB - Haitian women in Miami, Florida, responded to recruitment for testing of HIV antibody serostatus in ways that demonstrated the value of ethnographic methods for studying reactions to this kind of test, especially pre- and posttest counseling sessions. A total of 155 women between 14 and 61 years old, recruited in Miami in 1992 and 1993, participated. Response to testing identified three primary obstacles to the women's understanding of content presented in pre- and posttest counseling sessions: (1) their confusion about the meaning of positive versus negative, (2) the investigators' difficulty in communicating the concept of antibody, and (3) vagueness of the concept of window period between exposure and presentation of antibody. Retesting of a subset of Haitian participants helped to define sexual risk among these women in terms of having partners who had other partners and perception of supernatural risk. PMID- 9408788 TI - Patterns of onset and cessation of drug use over the early part of the life course. AB - This study uses retrospective drug use histories to examine the timing of drug use behavior among young people participating in a large random probability survey of residents of the province of Ontario, Canada. Results reveal that the major risk period for initiation into alcohol, tobacco, and most illicit drugs begins around age 12 and is mostly over by age 22. For most drugs, peak periods of risk of initiation occur in the age range 15 to 19. For all categories of illicit drugs, results reveal high quit rates in the first few years of use followed by a sharp decline. Differences by gender reveal higher rates of onset for males for most drugs. Male drug users are significantly less likely than female users to quit using a drug. Results are discussed in terms of their implications for the development of primary prevention and early intervention programs for adolescents and young adults. PMID- 9408789 TI - The telephone as a communication medium for health education. AB - The telephone is being widely used by the private sector as a communication medium for understanding and influencing consumer behavior. Coinciding with the growth of telephone use in the private sector is the expansion of telephone use in health care to include complex health promotion and disease prevention interventions aimed at initiating and maintaining health-related behavioral changes. While several studies have evaluated the impact of telephone interventions on a wide range of health behaviors, no published reports synthesizing current knowledge about using the telephone as a communication medium for health education were identified. In this article, the authors therefore (1) examine recent applications of telephone use in health education by describing three example interventions, (2) outline key features and alternatives in conceptualizing and designing health education using the telephone, (3) review advantages and disadvantages of using the telephone for health education, and (4) discuss implications for health education practice and research. PMID- 9408790 TI - The meaning of RTI in Vietnam--a qualitative study of illness representation: collaboration or self-regulation? AB - In collaboration with the National Committee for Population and Family Planning, a study was conducted in a rural and urban commune of northern Vietnam to provide community-level information about women's reproductive health and behaviors. Ethnographic and structured interviews were conducted with 32 women. A psychosocial model of health behavior, the Dual Process Model, was applied to provide a theoretical framework for understanding women's interpretations of, and strategies for, coping with symptoms of reproductive tract infections (RTIs). Women were found to interpret and manage RTI symptoms collaboratively with other women. Therefore, women's approach to care seeking was influenced heavily by their peer network and not driven by their method of family planning. PMID- 9408791 TI - Method effects in survey and focus group findings: understanding smoking cessation in low-SES African American women. AB - The same pool of African American women participated in a survey and in focus groups on motivation to quit smoking. Findings from the two studies were compared to explore potential method effects. Consistent with each method's basic purposes, the survey identified variation in study variables based on accepted theory and association patterns among such variables. The focus groups discovered themes and images salient to the participants and highlighted the situational contexts that gave meaning to smoking and smoking cessation. Survey method limitations included poor sensitivity to topic salience and contextual meanings and a deductive mode that channeled interpretation of results within the boundaries predefined by the study's theoretical framework. Focus group method limitations included an overfocus on the most dramatic and uncommon evidence and lack of systematic ways to identify explanations that may underlie the participants' overt expressions. Together, the multiple findings complemented and explained each other. PMID- 9408792 TI - Homogeneity of cigarette smoking within peer groups: influence or selection? AB - Many studies stress the relevance of peer group influence as a determinant of smoking behavior of adolescents. Recent research, however, concludes that homogeneity of behavior in friendships is also due to selection; youngsters choose new friends whose behavioral patterns are similar to their own. Data from a three-wave longitudinal study among students of secondary education (N = 1,063) was used to examine sources of peer similarities. The results demonstrated that both influence and selection processes contributed to peer group homogeneity, but the largest part of similarities in smoking status had to be attributed to selection. No support was found for friendships breaking down for reasons of dissimilarity in smoking status. Our findings underscore the complexity of processes and interactions with regard to the relationships of teenagers and offer new views on prevention programs. PMID- 9408793 TI - Evaluating community coalitions for prevention of substance abuse: the case of Project Freedom. AB - In the United States alone, there are more than 2,000 community coalitions to address local concerns about abuse of alcohol, tobacco, and other drugs. This article describes an evaluation system used to examine the process, outcome, and impact of coalitions for the prevention of substance abuse. The evaluation addresses five key questions: (a) Was the community mobilized to address substance abuse (Process)? (b) What changes in the community resulted from the coalition (Outcome)? (c) Is there a change in reported use of alcohol and other substances by youths (Outcome)? (d) Does the coalition have a community-level impact on substance abuse (Impact)? and (e) Is community-level impact related to changes facilitated by the coalition (Impact)? To address these and other questions, using eight core measurement instruments, the evaluation system collects 15 distinct measures. This evaluation system is illustrated with a multiyear study of Project Freedom, a substance abuse coalition in a large midwestern city. PMID- 9408794 TI - Depression and memory in the elderly: a meta-analysis. AB - A meta-analysis (N = 40) of the effects of depression on memory in the elderly (sample mean age > or = 55 years) examined variables potentially accounting for divergent findings in the literature. The distribution of effects was bimodal and the effect sizes were heterogeneous. Compared to controls, groups containing unipolar subjects only were significantly less impaired than were mixed unipolar bipolar; five of six studies mixing depression subtype were associated with the more negative mode. Samples containing younger depressed subjects (< 45 years) were significantly more impaired and were associated with the more negative mode. Significant group differences were found between studies matching their comparison groups reasonably well on years of education and those that did not. Thoroughness of dementia screening yielded no group differences. Although correlated observations precluded significance tests, larger effects were found for (1) figural (vs. verbal) memory; (2) delayed (vs. immediate) memory; and (3) recognition (vs. free recall and incidental or cued recall; incidental and cued recall effects were nearly identical). Similar effects were found for composite memory scores versus constituent and for various presentation paradigms (e.g., single presentation, selective reminding). Effect sizes for these categories were in the moderate range. Difficulties synthesizing this literature are discussed as are suggested remedies and directions for future research. PMID- 9408795 TI - Free and cued selective reminding and selective reminding in the elderly. AB - Free and Cued Selective Reminding (FCSR) differs from Selective Reminding (SR) because of a study procedure which controls cognitive processing and a reminding procedure which allows for cued recall. Performance on FCSR and SR was compared in two studies to determine which test produces better recall and to identify the factors that account for the superior recall. When the tests were administered to very elderly normal subjects using the standard clinical protocol, twice as many words were retrieved from long-term memory in FCSR than SR. The second experiment, which manipulated study and reminding conditions in a younger sample of normal elderly, suggests that the improvement in free recall was due to the study procedure and the method of reminding. PMID- 9408796 TI - Relationship of IQ to verbal learning and memory: test and retest. AB - The relationship between Wechsler Adult Intelligence Scale-Revised (WAIS-R) IQ and performance on measures of memory was examined in 64 adults tested twice at a 2-week interval. Repeated measures analyses of variance revealed that individuals with Low-Average WAIS-R Full Scale IQ scores performed significantly more poorly than did individuals with Average and High-Average Full Scale IQs on memory measures including the Wechsler Memory Scale-Revised (WMS-R) General Memory and Delayed Recall indices, as well as California Verbal Learning Test (CVLT) Total Words. Learning Slope, and Discriminability. Although all three groups demonstrated significant practice effects on each memory measure, group differences in performance persisted at retest. Multiple regression analyses revealed that WAIS-R factor scores Verbal Comprehension and Freedom from Distractibility accounted for up to 42% of the variance in WMS-R and CVLT indices. Moreover, WAIS-R performance at initial testing accounted for 22-41% of the variance in memory performance at retest. These results are discussed in the context of the construct stabilities of intelligence and memory, as well as the psychometric precision of the tests used to measure these constructs. PMID- 9408797 TI - Test-retest reliability for the consonant-vowel syllables dichotic listening paradigm. AB - The present study compared test-retest reliability for dichotic listening (DL) performance to consonant-vowel (CV) syllables under three different attentional instructions. Previous studies of reliability in dichotic listening have shown great variability which most likely reflects the large variations in stimuli, procedures, and techniques used in the different studies. Sixteen subjects were tested twice, 2 weeks apart, with the same dichotic listening procedure, including a divided attention condition and two focused attention conditions on each test occasion. The results showed test-retest correlations ranging from .61 to .86 for the three attentional conditions, with the smallest correlation coefficient for the divided attention condition. PMID- 9408798 TI - Free and cued selective reminding test: MOANS norms. AB - Age-adjusted norms are presented for persons age 56-98 years on the Free and Cued Selective Reminding Test (FCSRT; Buschke, 1984; Grober & Buschke, 1987). These data were obtained via several research projects that are known collectively as Mayo's Older Americans Normative Studies (MOANS). These norms should be useful when the FCSRT is used as a "stand alone" test. They should also promote accuracy in the comparison of any person's FCSRT performance against his or her functioning on any other tests with MOANS norms. The unique features of the FCSRT are discussed. These norms will, hopefully, enhance the utility of the test when it is used in clinical and research investigations of various amnestic disorders. PMID- 9408799 TI - Memory improvement following cardiac transplantation. AB - Seventeen patients with severe cardiomyopathy underwent neuropsychological evaluation prior to and at least 1 year after successful heart transplantation. Study candidates were screened, and individuals with a history of stroke, cardiac arrest, or medical and neurological conditions which might affect brain function were excluded. Pre-transplant testing revealed normal intelligence and normal attentional, language, and executive abilities but impaired recent memory. Following heart transplant, memory functioning improved significantly, reaching normal levels. Other cognitive abilities remained unchanged. Results suggest that cardiomyopathy is associated with mesial temporal dysfunction, possibly attributable to inadequate or reduced cerebral blood flow and related hypometabolism. This cerebral dysfunction is potentially reversible following successful transplantation, which restores cardiac output and cerebrovascular perfusion. PMID- 9408800 TI - Effects of triazolam and ethanol on proactive interference: evidence for an impairment in retrieval inhibition. AB - The effects of two memory-impairing drugs, ethanol and triazolam, on proactive interference (PI) in memory were studied. Following ingestion of either one of these drugs or a placebo, subjects studied an A-B list (e.g., BEE-WASP) of paired associates, followed by an A-C list (e.g., BEE-HONEY) on the interference trial, and a D-E list (e.g., KING-QUEEN) followed by an A-C list on the control trial. A PI effect was found in the data, such that subjects produced fewer correct second list targets on the interference trial than on the control trial. Neither ethanol nor triazolam was found to influence the size of the PI effect. However, both drugs were found to increase B intrusions on the test of the A-C list, to impair subjects' ability to produce more than one studied response for each cue word, and to impair the subjective experience of retrieved memory information. These data suggest that ethanol and triazolam impair an inhibitory process that normally operates as one component of intentional retrieval, playing an important role in the suppression of unwanted information during a memory task. PMID- 9408801 TI - Unmasking the heterogeneity of Alzheimer's disease: case studies of individuals from distinct neuropsychological subgroups. AB - Case descriptions of patients with probable Alzheimer's disease (AD) who were reliably classified into three neuropsychological subgroups (global [GAD], right hemisphere [RAD], and left-hemisphere [LAD]) in an earlier cluster analytic study (Fisher et al., 1996) are presented. Concordance between the neuropsychological patterns and clinical histories of randomly selected and hand-selected cases from within each subgroup was high. Longitudinal analysis revealed stable subgroup specific neuropsychological progression patterns. Results are discussed in terms of future research avenues worth pursuing, in addition to the conceptual shift in research design necessary to uncover both quantitative and qualitative subgroup specific ability differences. PMID- 9408802 TI - Confirmatory factor analysis of the Wechsler Memory Scale-Revised in a sample of clients with alcohol dependency. AB - Previous analyses of the Wechsler Memory Scale-Revised (WMS-R; Wechsler, 1987) have reported one-, two-, and three-factor solutions and raised questions about the validity of the visual memory subtests. These various findings may stem in part from different methods of analysis, and from the study of different participant samples. To address these issues, we analysed data from the WMS-R and a spatial maze test administered to 154 participants with a history of alcohol dependence. Results from confirmatory factor analysis supported the interpretation of three factors underlying the WMS-R subtests and the spatial maze score in this sample, namely, attention-concentration, immediate memory, and delayed recall. This result held despite the inclusion of the maze score which is a well-validated measure of visuo-spatial memory. PMID- 9408803 TI - Dissociation between position sense and visual-spatial components of hemineglect through a specific rehabilitation treatment. AB - Current evidence suggests an association between contralesional extra-personal hemineglect, and deficits of arm position sense in patients with damage to the right cerebral hemisphere. A unitary deficit may produce both disorders, or this association may reflect the anatomical contiguity of relevant brain structures. A rehabilitation treatment, devised for visual-spatial hemineglect, was used to investigate these hypotheses in 8 patients with damage to the right cerebral hemisphere. The treatment improved hemineglect, but not the position sense deficit. The severity of the latter was however transiently reduced by optokinetic stimulation, with effects similar to those found in visual-spatial hemineglect. These effects of rehabilitation suggest that extra-personal hemineglect and the neglect-related component of the position sense disorder of the left forearm are independent, though frequently associated, deficits. Implications for the design of rehabilitation programs are discussed. PMID- 9408804 TI - Solvent-induced toxic encephalopathy: electrophysiological data in relation to neuropsychological findings. AB - Male subjects with type 2A (n = 12) and 2B (n = 12) solvent-induced toxic encephalopathy and a reference group of healthy men (n = 12) without previous solvent exposure were studied using quantitative EEG and event-related potentials from an odd-ball and a dual-task paradigm. Subjects with toxic encephalopathy of types 2A and 2B showed markedly lower P300 amplitudes than did controls in both paradigms. In the relatively complex dual-task setting, subjects with 2A and 2B showed lower signal detection than did controls. PMID- 9408805 TI - Must we now consider SRIs neuroleptics? PMID- 9408806 TI - How long should pindolol be associated with paroxetine to improve the antidepressant response? AB - A double-blind study was undertaken to investigate the period of treatment with the beta-adrenoreceptor/5-hydroxytryptamine 1A (5-HT1A) antagonist pindolol required to enhance the antidepressant effects of paroxetine. After 1 week of a placebo run-in period, 63 untreated major depressive inpatients were randomly assigned to three different groups. Group 1 received paroxetine (20 mg/day) plus placebo (4 weeks). Group 2 received paroxetine (20 mg/day) plus pindolol (7.5 mg/day) for 1 week and placebo for 3 weeks. Group 3 received both active treatments for the entire duration of the study (4 weeks). Clinical response was defined as a reduction of the score in the Hamilton Rating Scale for Depression (HAM-D) to 8 or below. Also, to preliminarily examine whether beta-adrenoreceptor blockade was involved in the action of pindolol, another group of 10 inpatients was treated in an open-label manner with paroxetine (20 mg/day) plus 50 mg/day of the beta-adrenergic antagonist metoprolol, devoid of significant affinity for 5 HT1A receptors. At endpoint, the incidence of treatment-emergent side effects did not significantly differ among the three groups. After 1 and 2 weeks of treatment, the two groups treated with paroxetine plus pindolol displayed a significantly greater response rate than the group treated with paroxetine plus placebo. At study completion, only the patients treated with pindolol for the entire period showed a significantly greater response rate (p = 0.05). HAM-D score were also significantly lower at endpoint in patients treated with the combination for 4 weeks (p = 0.00003). The group of patients treated with paroxetine and metoprolol exhibited a side-effect profile comparable to that of paroxetine alone. Response rates were also comparable. These findings support the efficacy of pindolol, but not of metoprolol, in accelerating the antidepressant effect of paroxetine and suggest that the administration of pindolol for the entire period of the acute treatment may increase the efficacy of paroxetine. PMID- 9408807 TI - Paroxetine does not affect the cardiac safety and pharmacokinetics of terfenadine in healthy adult men. AB - Potent CYP3A4 inhibitors such as ketoconazole can cause dangerous increases in plasma concentrations of the H-1 antagonist terfenadine. In light of recent reports that the selective serotonin reuptake inhibitor antidepressants may be weak CYP3A4 inhibitors, this study was designed to investigate the effects of paroxetine on the pharmacodynamic and pharmacokinetic profile of terfenadine. Twelve healthy male volunteers participated in a randomized open-label, two period, steady-state crossover study. Terfenadine (60 mg twice daily for 8 days) was administered alone and with paroxetine at steady state (20 mg once daily for 15 days, with terfenadine on days 8 through 15). Extensive electrocardiogram monitoring was conducted throughout, and terfenadine and carboxyterfenadine pharmacokinetics were assessed at the end of each treatment period. One subject withdrew because of adverse experiences related to paroxetine, but the other 11 subjects completed the study uneventfully. On the final day of coadministration, the rate-corrected QT interval (QTc) was unaltered compared with terfenadine dosed alone; maximum QTc values (mean [SEM]) were 404 (4) and 405 (5) msec, respectively. Terfenadine pharmacokinetics were also unchanged; geometric mean steady-state area under the curve (AUC)tau values were 30.0 ng.hr/mL during coadministration compared with 30.8 ng.hr/mL when dosed alone (p > 0.05). The corresponding Cmax values were 3.68 and 3.64 ng/mL (p > 0.05). There was, however, a small (on average 17-20%), unexplained reduction in the steadystate Cmax and AUCtau of carboxyterfenadine during coadministration with paroxetine. In conclusion, paroxetine does not affect the pharmacokinetics or cardiovascular effects of terfenadine. The small reduction in carboxyterfenadine plasma concentrations is unlikely to be important clinically. PMID- 9408808 TI - General principles in the pharmacotherapy of antidepressant-induced rapid cycling: a case series. AB - The treatment of depression in patients who rapidly cycle once they receive antidepressant medication is a difficult clinical problem. Hurowitz and Liebowitz presented six patients with antidepressant-induced rapid cycling and proposed guidelines for how to manage medications in these patients. In this article, we present six new cases of antidepressant-induced rapid cycling as well as follow up from four of the previous cases. We conclude that DSM-IV criteria are not sufficiently sensitive to diagnose antidepressant-induced rapid cycling and that the treatment guidelines of Hurowitz and Liebowitz, although not foolproof, are useful. General recommendations for treating patients with antidepressant-induced rapid cycling are discussed. PMID- 9408809 TI - The CYP2D6 genotype and plasma concentrations of mianserin enantiomers in relation to therapeutic response to mianserin in depressed Japanese patients. AB - The relationship between therapeutic response to racemic mianserin and steady state plasma concentrations of S(+)- and R(-)-mianserin was studied in 26 Japanese patients with major depression. The daily dose of mianserin was 30 mg, and the duration of treatment was 3 weeks. Regarding S-mianserin, the proportion of responders (final Montgomery-Asberg Depression Rating Scale score of 10 or less) was significantly higher in the plasma concentration range of 10 to 23 ng/mL than outside (10 of 11 vs. 3 of 15, p = 0.0005). Such a plasma concentration difference between responders and nonresponders was not found for R mianserin. In 15 patients, the relationships between the CYP2D6 genotype, determined by allele-specific polymerase chain reaction analysis and Escherichia coli RI restriction fragment length polymorphism, plasma concentrations of the enantiomers, and the therapeutic response were studied. Five patients were homozygous for the wild type (wt) allele (wt/wt), nine were heterozygous for the CYP2D6Ch (Ch) allele causing decreased CYP2D6 activity (Ch/wt), and one patient was heterozygous for the Ch allele and the defect allele CYP2D6D (D) (Ch/D). The Ch/wt group showed significantly higher plasma concentrations of S-mianserin (mean +/- SD: 15 +/- 6 vs. 8 +/- 1 ng/mL, p = 0.007) and proportion of responders (8 of 9 vs. 1 of 5, p = 0.023) than the wt/wt group. The patient with the Ch/D genotype had the highest plasma concentration of S-mianserin (37 ng/mL) and a poor response. No significant relationship was found between the CYP2D6 genotype and plasma concentration of R-mianserin. The study presented here thus suggests that the CYP2D6 genotype plays a major role in controlling plasma concentration of the S-enantiomer of mianserin, which contributes to a major extent to the antidepressant effect during mianserin treatment. PMID- 9408810 TI - Olanzapine plasma concentrations and clinical response in acutely ill schizophrenic patients. AB - Olanzapine is an atypical antipsychotic effective in the treatment of schizophrenic patients. After a 2- to 9-day placebo lead-in, 79 inpatients with schizophrenia according to DSM-III-R criteria were placed on an olanzapine dosage of 10 mg/day or 1 mg/day for up to 6 weeks. Blood samples were obtained weekly during this period. Receiver operating characteristic curve analyses of Brief Psychiatric Rating Scale (BPRS) and Positive and Negative Syndrome Scale rating scale data suggested a minimum effective therapeutic concentration of 9 ng/mL. Using an intent-to treat analysis, 45% of the patients with olanzapine plasma concentrations > or = 9.3 ng/mL responded (> or = 20% decrease in BPRS), whereas only 13% of the patients with concentrations < 9.3 ng/mL responded. Use of olanzapine plasma concentrations of > 9 ng/mL as a predictor for treatment response in acutely ill schizophrenic patients is practicable because this therapeutic marker significantly increases the likelihood of a patient responding to olanzapine. PMID- 9408811 TI - Risperidone dose and blood level variability: accumulation effects and interindividual and intraindividual variability in the nonresponder patient in the clinical practice setting. AB - Risperidone blood levels were measured every 2 weeks after initiation of therapy in 24 refractory chronic schizophrenic patients referred to a locked, skilled nursing facility for long-term treatment. Blood levels were assessed on 285 occasions over a 1- to 16-month treatment program. Drug plasma level increases peaked by 2 months for risperidone at 334% and by 6 months for 9-hydroxy risperidone at 104% over the baseline levels. Total blood levels (risperidone plus 9-hydroxy-risperidone) peaked at 111% increase at 6 months and then declined 8% per month to 12 months, stabilizing at a value 31% higher than the initial value. Significant dose to blood level interindividual variation was noted. Considerable blood level variation was evident in single blood level sample determinations. The results suggest the value of risperidone blood levels, consideration of reduction of initial recommended starting dosages, and a need to optimize risperidone dosage approaches individually to patients. PMID- 9408812 TI - Replacement medication for cocaine dependence: methylphenidate. AB - Agonists, or "replacement medications," are useful adjuncts in treatment of opiate and nicotine dependence. They have not been systematically examined in cocaine dependence. Results of early open trials with methylphenidate for treatment of cocaine dependence were equivocal. Twenty-four cocaine-dependent subjects were enrolled in an 11-week double-blind, placebo-controlled study of methylphenidate. Assignment was random. Intake included a 2-day human laboratory procedure in which subjects received initial doses of methylphenidate or placebo. Subjects attended the clinic Monday through Friday and received oral doses of methylphenidate (5 mg plus 20-mg sustained release) or placebo at 8:00 a.m., with afternoon and weekend take-home doses (20 mg sustained-release or placebo) provided in Medication Events Monitoring System bottles to monitor compliance. Clinic visits included therapy sessions, electrocardiograms, self-report measures, and twice-weekly urine screens. The two groups were equivalent in terms of retention (methylphenidate 48% and placebo 42%) and had similar cocaine use outcomes (40% benzoylecgonine-positive urine screens). There were no significant adverse effects. The doses were sufficient to permit detection of psychoactive effects ("stimulant," "more energy") and side effects ("jitteriness," "eating less") without increased "craving." Additional medications with different effects profiles are being studied to further evaluate the replacement model in cocaine dependence. PMID- 9408813 TI - Selective serotonin reuptake inhibitor and akathisia. PMID- 9408814 TI - Clinical considerations in managing nausea associated with venlafaxine. PMID- 9408815 TI - Seizure activity and enzyme elevations after venlafaxine overdose. PMID- 9408816 TI - Serotonin, cigarettes, and nausea. PMID- 9408817 TI - A naturalistic pilot study comparing haloperidol, clozapine, sertindole, and risperidone in partially responsive chronic schizophrenia or schizoaffective disorder. PMID- 9408818 TI - Prolongation of clozapine-induced granulocytopenia associated with olanzapine. PMID- 9408819 TI - End-stage renal disease after treatment with lithium. PMID- 9408820 TI - Botulinum toxin in the treatment of tardive dystonia. PMID- 9408821 TI - A linear goal programming model for human resource allocation in a health-care organization. AB - This paper presents the development of a goal programming (GP) model as an aid to strategic planning and allocation for limited human resources in a health-care organization. The purpose of this study is to assign the personnel to the proper shift hours that enable management to meet the objective of minimizing the total payroll costs while patients are satisfied. A GP model is illustrated using the data provided by a health-care organization in the midwest area. The goals are identified and prioritized. The model result is examined and a sensitivity analysis is performed to improve the model applicability. The GP model application adds insight to the planning functions of resource allocation in the health-care organizations. The proposed model is easily applicable to other human resource planning process. PMID- 9408822 TI - A process model basis for evolving hospital information systems. AB - Hospitals often invest significant resources in the development of large and complex information systems that must be modified and extended to respond to changing requirements and to exploit the capabilities offered by modern technologies. A disciplined approach of managing the evolution of hospital information systems is then required so that to meet the increasing demands for effective and efficient use of scarce resources. This paper takes a process oriented view of the hospital and presents an approach based on hospital process modeling which aims at assessing the current status of computer support within a hospital and at identifying new opportunities for automation. The approach is illustrated by an example taken from a major Greek hospital. PMID- 9408823 TI - Graphical 3D medical image registration and quantification. AB - We present a graphical three-dimensional method that facilitates image registration and fusion, and provides quantitative geometric and volume information. In particular it enhances the use of functional (radiopharmaceutical) imaging (SPECT, PET) which, though a powerful clinical tool, has the disadvantage of low spatial resolution and ill-defined boundaries. Registration between functional images and structural images (MRI, CT) can augment the anatomical context of these functional images. PMID- 9408824 TI - A human resource planning model for hospital/medical technologists: an analytic hierarchy process approach. AB - This paper deals with the development of a human resource planning (HRP) model for hospital laboratory personnel. External and organizational factors impacting the demand and supply of clinical laboratory personnel in an urban academic health center are identified by a Delphi process. These factors are structured into a hierarchy for the application of the Analytic Hierarchy Process (AHP). Computer software, Expert Choice, was used in model hierarchy development and judgmental elicitation process. The model result can be applicable to eliciting the perceptions, insights, and understanding of health-care decision-makers for strategic human resource planning. PMID- 9408825 TI - Evaluation of an algorithm to identify women with carcinoma of the breast. AB - Although claims data are increasingly being used to measure and manage the cost and quality of health care, few studies have evaluated algorithms developed for such analyses. Therefore, the present study was performed to evaluate prospectively a previously published algorithm used to identify women with the new diagnosis of carcinoma of the breast. This algorithm had been developed from the patterns of claims that suggested common clinical presentations of carcinoma of the breast. In the present study, this algorithm was used to identify 177 potential cases of women with newly diagnosed carcinoma of the breast from the claims database of a large health maintenance organization (HMO). The algorithm's positive predictive value for cases identified in the present study was 83% (147/177). To attempt to improve upon the positive predictive value, multiple modifications of the algorithm were performed. The previously defined best modification of the initial algorithm yielded a positive predictive value of 84% (147/174) in the present study with the loss of none of the true positive cases. These results demonstrate that logic-based algorithms can be used as a valid and efficient method of identifying large numbers of new breast cancer cases from claims data. This algorithm provides a powerful tool to perform health care analysis and research for women with newly diagnosed carcinoma of the breast. PMID- 9408826 TI - Optical biosensors. PMID- 9408827 TI - The influence of the exocyclic amino group characteristic of GC base pairs on molecular recognition of specific nucleotide sequences in DNA by berenil and DAPI. AB - The expedient of preparing homologous DNA samples substituted with inosine for guanosine residues, 2,6-diaminopurine (DAP) for adenine residues, or both, has been used to investigate the role of the purine 2-amino group in determining the preferred binding sites for the drugs berenil [1,3-bis(4-phenylamidinium) triazene] and DAPI (4',6-diamidino-2-phenyl indole) on DNA. The selectivity of these two minor groove binders for AT-rich sequences is seen to be radically altered in the substituted DNA molecules. Neither berenil nor DAPI bind to DAP substituted DNA where all purine residues bear a 2-amino group. By contrast, they bind to AT-rich, IC-rich and even mixed sequences of the inosine DNA where all purine residues lack the 2-amino group. With the inosine and DAP double substituted DNA, both berenil and DAPI bind preferentially to IC-rich clusters instead of their canonical tracts endowed with an extra 2-amino group through substitution with DAP. These results establish that the location of the purine 2 amino group represents a critical determinant for recognition of DNA nucleotide sequences by the two drugs. PMID- 9408828 TI - Ligand loading at the surface of an optical biosensor and its effect upon the kinetics of protein-protein interactions. AB - Optical biosensors are finding increasing use in the determination of kinetic and equilibrium constants for a variety of biomolecular interactions. Usually these biosensors require one biomolecule, the ligand, to be covalently attached to a hydrogel matrix which itself is bonded to the sensing surface. The ligands partner, the ligate, then binds from solution resulting in a measurable change in response which the instrument records as a function of time. Although in many cases, optical biosensors are used in order to obtain parameters that relate to interactions in solution, it is becoming clear that measurements involving the interaction of ligate with immobilized ligands on surfaces require careful experimental design. Here we report on how the density of ligand loading within the hydogel matrix affects the measured interaction kinetics. It is found that crowding of ligand within this matrix results in a significant reduction in the measured association rate constant, with a corresponding effect in the calculated overall affinity. However, measurements at low ligand loadings show association rate constants that are comparable to those measured in solution. Clearly, where this comparison is required, it is important to perform measurements under such conditions. PMID- 9408829 TI - Detection and characterization of weak affinity antibody antigen recognition with biomolecular interaction analysis. AB - In biological systems, weak-affinity interactions (association constant, Ka, of less than approximately 10(4) M-1) between biomolecules are common and essential to the integrity of such units. However, studies of weak biological interactions are difficult due to the scarcity of analytical methods available for the bioscientist. In this communication, we report on the use of biosensors based on surface plasmon resonance to detect and characterize weak affinity antibody antigen interactions. Monoclonal antibodies towards carbohydrate antigens were immobilized on sensor surfaces and were used to detect weak binding of the carbohydrate tetraglucose of dissociation constant, Kd, in the millimolar range. Sensorgrams were received in the form of square pulses where the kinetic rate constants were difficult to assess due to the rapid association and dissociation of the antigen to/from the immobilized antibody. PMID- 9408830 TI - Further refinement of the description of the ligand-binding characteristics for the galactoside-binding mistletoe lectin, a plant agglutinin with immunomodulatory potency. AB - The galactoside-binding lectin from mistletoe (Viscum album L.) is a biological response modifier, eliciting e.g. enhanced secretion of cytokines. This immunological activity warrants the further analysis of its ligand-binding properties with special attention paid to blood group epitopes. To avoid the microheterogeneity and complexity of naturally occurring glycoproteins, chemically strictly defined neoglycoconjugates and a panel of synthetic oligosaccharides were employed in solid-phase assays for direct binding and assessment of the relative inhibitory capacity. Since label incorporation into the lectin, although performed under protective conditions, or surface immobilization by adsorption to plastic may affect its affinity characteristics, the extent of neoglycoconjugate binding in the absence of any interfering substance and in the presence of oligosaccharides was determined comparatively with labeled and with immobilized lectin. In principle, these two factors could be excluded to markedly alter binding features. In addition to lactose, the blood group determinants H and B were strongly reactive. A fucose residue can thus especially be accommodated to the binding site when linked to the non-reducing unit. N-Acetyllactosamine was nearly as potent as an inhibitor as lactose. Lec and the A determinant were notably inferior to the other ABH blood group epitopes. Le(a) and Le(x) and their sialylated derivatives displayed only very weak binding capacity. Among the two natural isomers of sialyllactose, the alpha 2,6-form displayed a higher level of inhibitory capacity than the alpha 2,3 derivative. Isomeric variants of the Thomsen-Friedenreich antigen, too, reduced lectin binding to the lactose-carrying polymer. Their capacities were surpassed by those of the H and the B determinants and a related form of the latter, the P1 epitope. An overlap of specificity with the immunomodulatory human galectin-3 is thus measurable for H/B-like structures. The documented differential reactivity of the mistletoe lectin to blood group oligosaccharides may have a bearing on the responsiveness of blood group-positive cell populations. PMID- 9408831 TI - Influence of mass transfer and surface ligand heterogeneity on quantitative BIAcore binding data. Analysis of the interaction of NC10 Fab with an anti idiotype Fab'. AB - The interaction of monovalent Fab fragments of NC10, an antiviral neuraminidase antibody, and the anti-idiotype antibody 3-2G12 has been used as a model system to demonstrate experimentally the influence of non-ideal binding effects on BIAcore binding data. Because the association rate constant for these two molecules was found to be relatively high (about 5 x 10(5) M-1 S-1), mass transfer was recognised as a potential source of error in the analysis of the interaction kinetics. By manipulation of the flow rate and the surface density of the immobilised ligand, however, the magnitude to this error was minimised. In addition, the application of site-specific immobilisation procedures was found to improve considerably the correlation of experimental binding data to the ideal 1:1 kinetic model such that the discrepancy between experimental and fitted curves was within the noise range of the instrument. Experiments performed to measure the equilibrium constant (KD) in solution resulted in a value of similar magnitude to those obtained from the ratio of the kinetic rate constants, even those measured with a heterogeneous ligand or with a significant mass transfer component. For this system, the experimental complexities introduced by covalent immobilisation did not lead to large errors in the KD values obtained using the BIAcore. PMID- 9408832 TI - Bioavailability of different artemisinin tablet formulations in rabbit plasma- correlation with results obtained by an in vitro dissolution method. AB - The demonstration of a good overall correlation with in vivo data is the ultimate proof of qualification for any dissolution-rate test. For artemisinin, a very hydrophobic compound at a high content in oral solid dosage forms, all official dissolution apparatus were estimated unsuitable. A modified two phase partition dissolution method was applied to solve this problem. This study reports on the bioavalability of three different formulations of artemisinin tablets in rabbit plasma. Artemisinin concentrations in plasma were determined by liquid chromatography. A linear correlation between results obtained by the partition dissolution method described and the obtained in vivo data confirmed the validity of the dissolution method. PMID- 9408833 TI - Increasing urinary cotinine concentrations at elevated temperatures: the role of conjugated metabolites. AB - The presence of cotinine, a nicotine metabolite, in urine above a specified cutoff concentration is commonly used to distinguish smokers from nonsmokers, as in smoking cessation studies. A stability study of cotinine in urine was carried out after questions arose concerning analyte stability at elevated storage and shipment temperatures. Aliquots from a smokers urine pool were stored at 5, 25, 40, 50 and 60 degrees C for 30 days. Another set of aliquots, obtained by diluting the smokers pool 1:1 with nonsmokers urine, were stored under the same conditions. Free cotinine levels, determined by a stability-indicating gas chromatographic/mass spectrometric (GC/MS) assay, increased over the 30-day period at higher storage temperatures. Cotinine concentrations in the aliquots stored at 60 degrees C, for example, nearly doubled over 30 days (1301 to 2476 ng/ml), with similar proportional increases observed in the aliquots diluted with nonsmokers urine. Since cotinine can be excreted to a large degree as cotinine-N glucuronide, this conjugated metabolite was determined by an indirect method. As the storage temperature increased, the free/conjugated cotinine ratio dramatically increased, pointing to cotinine-N-glucuronide as the source of the additional free cotinine at the higher storage temperatures. The results of this study are of much practical importance, since urine samples with free cotinine concentrations just below a cutoff value may become positive for smoking status if suitably low temperatures cannot be maintained during sample handling and shipment. PMID- 9408834 TI - Stability and compatibility of topotecan hydrochloride for injection with common infusion solutions and containers. AB - The stability and compatibility of topotecan hydrochloride with common infusion solutions and containers were studied. During this study, the leaching of diethylhexyl phthalate (DEHP), a major plasticizer of some polyvinyl chloride (PVC) materials was also investigated. A formulation of topotecan hydrochloride was added to 50 ml PVC infusion bags, polyolefin infusion bags and 150 ml glass bottles containing either 5% dextrose injection or 0.9% sodium chloride injection at an initial nominal topotecan concentration of 0.05 mg ml-1. Additionally, the topotecan hydrochloride formulation was added to 50 ml PVC infusion bags containing either 5% dextrose injection or 0.9% sodium chloride injection at an initial nominal topotecan concentration of 0.025 mg ml-1. Containers were maintained at 5 degrees C for 7 days or 23-24 degrees C for 24 h. Samples were analyzed using a stability-indicating HPLC method to determine the concentration of topotecan and the presence of any degradates. The samples were also analyzed by separate HPLC methods to detect the presence of DEHP and the hydrolyzed lactone ring form (SKF 105992) of topotecan hydrochloride. In addition, the pH of each sample was measured initially and at the end of the storage time. There was no significant loss of topotecan observed for any of the conditions studied and no significant increase in degradates was observed. The pH remained unchanged for all samples between the start and end of the study. At the concentrations studied, topotecan hydrochloride was stable for up to 24 h at room temperature and for up to 7 days at 5 degrees C, in PVC and polyolefin infusion bags and glass bottles containing either 5% dextrose injection or 0.9% sodium chloride injection. The presence of topotecan hydrochloride did not contribute to leaching of DEHP in the PVC infusion bags. PMID- 9408836 TI - Determination of glafenine in dosage forms and serum by thin layer densitometry and high performance liquid chromatography. AB - New thin layer densitometry and high performance liquid chromatography (HPLC) methods are described for quantitative determination of glafenine in dosage forms in the presence of its photo-degradation products and in serum in the presence of its metabolites. Mobile phases consisting of toluene-isopropyl alcohol dimethylformamide-water (18:3:1:0.5) and methanol-water-phosphoric acid (80:120:0.5) are found to be efficient for reasonable separation and adequate resolution of glafenine from associated substances by thin layer chromatography (TLC) and HPLC techniques, respectively. The methods are used for the study of glafenine purity, stability, bioavailability, bioequivalence and tablet dissolution rate. The results obtained by TLC and HPLC techniques are in good agreement and offer the advantages of reproducibility and accuracy. PMID- 9408835 TI - Analysis of amino acid enantiomers derived from antitumor antibiotics using chiral capillary electrophoresis. AB - The chiral separation of enantiomeric forms of derivatized amino acids have been achieved based on a metalchelate chiral capillary electrophoretic method and a cyclodextrin mediated host-guest interaction approach in micellar electrokinetic chromatography (MEKC) mode with laser-induced fluorescence detection. This approach has been applied to the determination of enantiomeric forms of amino acids derived from novel depsipeptide antitumor antibiotics, BMY-45012 and its analogs. Amino acids were analyzed by complete hydrolysis and the hydrolysate was derivatized with either dansyl chloride for UV absorbance detection or fluorescein isothiocyanate for laser based fluorescence detection. The presence of several amino acids, serine and beta-hydroxyl-N-methy-valine in the proposed structure have been confirmed as D-serine and L-beta-hydroxyl-N-methy-valine enantiomeric forms by both chiral capillary electrophoresis (chiral CE) and MEKC approaches. A non-chiral amino acid, sarcosine, was also confirmed. These methodologies provide a quick and sensitive approach for the determination of amino acids racemization of pharmaceutical natural products and have proven to be useful for structural elucidation refinement. PMID- 9408837 TI - Relationship between lipophilicity and binding to human serum albumin of arylpropionic acid non-steroidal anti-inflammatory drugs. AB - A possible relationship between lipophilicity and binding to human serum albumin was investigated for 11 arylpropionate non-steroidal anti-inflammatory drugs. The lipophilic parameter was determined by a reversed-phase high-performance liquid chromatographic procedure as the capacity factor (k'). The binding of arylpropionic acids to human serum albumin was studied in vitro by equilibrium dialysis. For each compound, a Scatchard analysis was performed considering two classes of binding sites characterized by high- and low-affinity constants, K1 and K2, respectively. A linear relationship was found between lipophilicity and binding parameters, n1K1 (r = 0.88, P < 0.0005) and n2K2 (r = 0.96, P < 0.0002). These results suggest the role of hydrophobic interactions in the binding of arylpropionic acids to human serum albumin. PMID- 9408838 TI - Optimisation of the enantiomeric separation of 12 2-aminotetralin analogues using Chiral AGP high-performance liquid chromatography by simultaneous factorial design. AB - A method for the simultaneous optimisation of mobile phase composition for the resolution of pairs of enantiomers of 12 2-aminotetralin analogues is presented. The selectivity necessary to discriminate between 12 analytes was obtained by using mass selective detection. The ability to examine more than a few analytes at a time extends the otherwise limited applicability of a factorial design strategy to the rapid development of chiral assays. PMID- 9408839 TI - Application of microdialysis to study the in vitro metabolism of drugs in liver microsomes. AB - Current methods for studying in vitro drug metabolism involve add-incubate separate-measure approach. Separation of the desired analytes requires removal of protein which is typically accomplished by precipitation and centrifugation and extraction of the analytes into an organic phase. The analysis scheme then becomes more complex resulting in a decrease in precision and an increase in assay time. Microdialysis sampling circumvents these problems by allowing researchers to sample the reaction mixture periodically and obtain the complete metabolic profile. In the present study, microdialysis sampling was used to investigate Phase I metabolism of salicylic acid, diazepam and ibuprofen in rat liver microsomes. The major metabolites of these drugs were profiled by LC. Michaelis-Menten enzyme kinetic parameters, Km and Vmax were obtained for the formation of diazepam metabolites by both microdialysis and conventional microsomal incubations and were in good agreement with the values reported in the literature. This study shows that microdialysis has considerable promise as a sampling technique for in vitro drug metabolism studies. By making minor modifications to the instruments, microdialysis can be applied to other in vitro systems such as isolated hepatocytes to study the Phase II metabolism or tissue slices to study drug distribution. PMID- 9408840 TI - Flow extraction spectrophotometric method for the determination of diclofenac sodium in pharmaceutical preparations. AB - The spectrophotometric determination of trace amounts of diclofenac was carried out by liquid-liquid extraction using acridine yellow with a flow system. The determination of diclofenac sodium in the range of 3-80 micrograms ml-1 was possible with a sampling frequency of 40 samples h-1. The method was satisfactorily applied to the determination of diclofenac in pharmaceutical preparations. PMID- 9408841 TI - Determination of water in ferrous lactate by near infrared reflectance spectroscopy with a fibre-optic probe. AB - Near infrared diffuse reflectance spectroscopy with a fibre-optic probe was used to determine the water content in ferrous lactate dihydrate. Spectra were recorded by immersing the probe in a beaker containing the ferrous lactate sample. Spectral data were processed by using two different multivariate calibration procedures, viz. stepwise multiple linear regression (SMLR) and partial least-squares regression (PLSR). The results provided by the two calibration procedures were similar and departed by less than 1.5% from the values obtained by Karl Fischer titration. PMID- 9408842 TI - Novel non-acidic formulations of haloperidol complexed with beta-cyclodextrin derivatives. AB - Haloperidol (Hal), a highly hydrophobic drug, was complexed with two beta cyclodextrin (beta-CD) derivatives. Hal solubility was increased 20-fold in the presence of a 10-fold excess of methyl beta-CD (Me beta-CD) and 12-fold in the presence of a 10-fold excess of 2-hydroxypropyl beta-CD (HP beta-CD). The stoichiometries and stability constants of Hal-Me beta-CD (1:1 and 2345 M-1 at 27 degrees C) and Hal-HP beta-CD (1:1 and 2112 M-1 at 27 degrees C) complexes were calculated by the continuous variation and phase solubility methods respectively. Differential scanning calorimetry and 1H-NMR were used to confirm the formation of inclusion complexes. Moreover, the enthalpy and entropy of the complexation process were calculated for both complexes in order to obtain such information as the main 'driving force' and whether or not complex formation is thermodynamically favoured. This was achieved by monitoring the isothermic solubility lines at various temperatures. PMID- 9408843 TI - Use of integral and differential methods for the determination of L-dopa in pure form and pharmaceutical preparations. AB - A new simple and selective kinetic method for the determination of L-dopa, using differential and integral methods, is described. The spectrophotometric measurements were recorded by measuring the increase in absorbance at 300 nm. The concentration range was valid from 5-35 ppm. The complex ratio showed a (1:2) of L-dopa with respect to sodium hydroxide, with formation constant 5.06 x 10(6) and molar absorptivity of 3.85 x 10(3) l mol-1 cm-1. PMID- 9408844 TI - Evaluation of the methods for the determination of the stability constant of cyclodextrin-chlorambucil inclusion complexes. AB - The interaction of the antitumor agent chlorambucil (CHL) with three different cyclodextrins (CD), namely methyl-beta CD (Me beta CD) polymer-beta CD (poly-beta CD) and gamma CD, is examined kinetically and spectrophotometrically, monitoring the hydrolysis and the changes in the UV absorbance of CHL respectively, in the presence of increasing concentrations of the examined CD. The stoichiometry coefficient for all the CHL-CD complexes was calculated and found to be 1:1, using the continuous variation method based on the UV data. Also, the stability constant Kst for the CHL-CD complexes was calculated and evaluated using the above mentioned two methods, each one based on linear and nonlinear mathematical models. All studies demonstrate that the interaction of CHL with the methylated derivative (Me beta CD) is stronger (the highest Kst value), probably due to the enhanced hydrophobic character of this derivative. PMID- 9408845 TI - The indirect UV detection in the analysis of ursodeoxycholic acid and related compounds by HPCE. AB - A high-performance capillary zone electrophoretic (HPCE) assay has been developed for the determination of ursodeoxycholic acid (UDCA) and its usual impurities. Considering the low molecular absorptivity of UDCA and its related compounds indirect UV detection was used. The electrophoretic capillary was filled with a background electrolyte (BGE) containing an UV absorbing ion: benzoic acid (BA) or 5,5-diethylbarbituric acid (DBA). To enhance the selectivity of the assay diimethyl-beta-cyclodextrines (D-beta-CDs) or trimethyl-beta-cyclodextrines (T beta-CDs) have been added to the running buffer together with methylcellulose or urea. All considered impurities were well resolved with two buffers studied, with the exception of methylursodehoxycholate, a neutral compound. PMID- 9408846 TI - Direct injection HPLC method for the determination of phenylbutazone and oxyphenylbutazone in serum using a semipermeable surface column. AB - A direct injection HPLC method has been developed for the determination of phenylbutazone and its active metabolite oxyphenylbutazone in serum using a semipermeable surface (SPS) column. The method is easy to perform and requires 20 microliters of a filtered serum sample. The chromatographic time is less than 13 min using a mobile phase of 15:85 v/v acetonitrile-0.05M phosphate buffer pH 7.5. The method was linear in the range 0.5-20 micrograms ml-1 (r > 0.99, n = 6) with R.S.D. less than 6%. Interday and intraday variability were found to be less than 8.3%. The limit of quantitation and detection were 0.5 and 0.25 microgram ml-1 (s/n > 3), respectively, for both drug and metabolite. PMID- 9408847 TI - Photon correlation spectroscopy applied to characterisation of denaturation and thermal stability of human albumin. AB - Photon correlation spectroscopy and light absorption measurements have been applied for characterisation of denaturation kinetics and thermal stability of human albumin in solution. The hydrodynamic size of the molecules has been studied as a function of pH, and the denaturation rate of ten different lots of 5% (w/v) human albumin solution has been measured at various temperatures. In the native (pH 7) state, the hydrodynamic molecular diameter was found to 6.3 nm. The molecular size was relatively stable between pH 10 and 5, but increased with decreasing pH to approximately 20 nm at pH 3. The denaturation rate, measured as change in hydrodynamic diameter per min, was strongly dependent on temperature and increased 3-fold per degree in the 73-75 degrees C range. The investigated lots of albumin solution showed large variations in stability at 74 degrees C, with denaturation rates ranging from 10 to 100 nm min-1. The observed thermal stability for the lots investigated was ranked identically with both the employed techniques. In an effort to explain the observed lot to lot variations in denaturation rate, a broad chemical characterisation including determination of free SH content, fatty acid content and composition and metal content, was performed. However, lot to lot variations in these parameters was not found to fully elucidate the observed variations in thermal stability. PMID- 9408848 TI - Quantitation of promethazine enantiomers in human serum using a chiralcel OJ-R column and mixed-mode disc solid-phase extraction. AB - A liquid chromatographic method was developed for the assay of R(+)- and S(-) promethazine enantiomers from human serum. The method involves the use of the mixed mode disc solid-phase extraction technique for sample clean-up. Chromatographic resolution of the enantiomers was performed on a reversed-phase cellulose-based chiral column (Chiralcel OJ-R) under isocratic conditions using a mobile phase consisting of 0.5 M aqueous sodium perchlorate/acetonitrile (63:37, v/v) at a flow rate of 0.5 ml min-1. Recoveries in the range of 97-99% at 20 ng ml-1 levels were obtained for both promethazine enantiomers. Intra-day and inter day precision calculated as R.S.D.% was in the 3-8% ranges for both enantiomers. Intra-day and inter-day accuracy calculated as percent error was in the 0-10 and 1-7% ranges for both enantiomers, respectively. Linear calibration curves were obtained for each enantiomer in serum in the concentration range 5-90 ng ml-1. The limit of quantitation of each enantiomer was 10 ng ml-1. The detection limit for each enantiomer in serum using UV detection at 249 nm was 2 ng ml-1 (S/N = 2). PMID- 9408849 TI - Assay for quantitation of clozapine and its metabolite N-desmethylclozapine in human plasma by high-performance liquid chromatography with ultraviolet detection. AB - A high-performance liquid chromatographic method with UV detection has been developed for the analysis of clozapine and its active N-desmethylated metabolite (N-desmethylclozapine = DMC) in human plasma. A liquid/liquid procedure was used to extract clozapine and DMC from human plasma. The analysis was performed on a C8 Nucleosil column and the mobile phase comprised acetonitrile-water-Pic B5 diethylamine (63:37:25:0.04, v/v/v/v). The detection wavelength was 245 nm. The intra-assay and inter-assay precision was satisfactory within the concentration range 10-900 ng ml-1. The lower detection limit for clozapine and for DMC was 5 ng ml-1. The recovery and reproducibility values of this method were better or similar to those found by other authors. This method, which is simple, selective and avoids an evaporation step, can be used routinely for therapeutic drug monitoring. PMID- 9408850 TI - The preparation of a molecular imprinted polymer to 7-hydroxycoumarin and its use as a solid-phase extraction material. AB - A molecular imprinted polymer (MIP) was prepared to 7-hydroxycoumarin (7-OHC). A number of preparation parameters were examined by ultraviolet (UV) spectroscopy, including the amount of solvent used for reaction, equilibration time, selectivity and capacity of material. The polymer which showed the most selectivity for 7-OHC was then packed into cartridges and used as a solid-phase extraction sorbent. An extraction procedure was then developed from first principles. The cartridges were examined for selectivity of 7-OHC over some other members of the coumarin family. 7-OHC was then extracted from urine using this solid-phase extraction (SPE) method, and its concentration determined using capillary zone electrophoresis (CZE). The method was found to be linear over the range 10-50 micrograms ml-1. Inter- and Intra-assay precision studies were performed to validate the method. PMID- 9408851 TI - New minor ecdysteroids from Silene otites (L.) Wib. AB - Six minor new ecdysteroid components have been isolated from Silene otites (L.) Wib. by a combination of chromatographic methods. Three of them (2-deoxy-20 hydroxyecdysone 3,22-diacetate, 5 alpha-2-deoxy-20-hydroxyecdysone 3-acetate, and 2-deoxy-20-hydroxyecdysone 3-crotonate) are new natural products. PMID- 9408852 TI - Determination of omeprazole in pharmaceuticals by derivative spectroscopy. AB - A new derivative UV spectroscopic method was developed for the analysis of omeprazole in borate buffer (pH 10.0; 0.1 M). Second derivative spectra were generated between 200-400 nm at N = 9, delta gamma = 31.5. The linearity range for values obtained from second derivative spectra was 0.2-40.0 micrograms ml-1. The developed method was applied to five different commercial preparations of hard gelatin capsules containing enteric coated granules. The relative standard deviations were found to be 2.24% (brand A), 1.87% (brand B), 2.80% (brand C), 4.55% (brand D) and 1.09% (brand E). The data were compared with ones obtained from the polarographic method given in the literature and no difference was found statistically. PMID- 9408853 TI - Determination of platinum in tumour tissues after cisplatin therapy by electrothermal atomic absorption spectrometry. AB - A simple procedure is described for the determination of platinum in tumours after cisplatin therapy. Tumours were digested in 65% nitric acid by incubation at 37 degrees C for 2 days and platinum analysed under optimum conditions by electrothermal atomic absorption spectrometry with Zeeman background correction. The reproducibility of measurements in general was better than +/- 2%. The calibration graph was linear from 30.0 up to 1000 micrograms l-1 of platinum, while the limit of detection (3 sigma) was found to be 3.0 micrograms l-1 (sample volume 20 microliters). Aqueous standard solutions and the standard addition method were applied in the calibration procedure. Under the recommended analytical conditions, the sample matrix did not influence the determination of platinum significantly. In 72% of samples analysed the differences between results obtained by the two calibration procedures did not exceed +/- 5%. PMID- 9408854 TI - Application of the bivariate spectrophotometric method for the determination of metronidazole, furazolidone and di-iodohydroxyquinoline in pharmaceutical formulations. AB - The bivariate calibration algorithm was applied to the spectrophotometric determination of metronidazole, furazolidone and di-iodohydroxyquinoline in pharmaceutical dosage forms. The results obtained were compared with the results of derivative spectrophotometry. The statistical evaluation of method bias was carried out, and it was shown that the proposed procedure may be competitive with commonly used first-derivative spectrophotometry. The advantage of the bivariate calibration is its simplicity, and the fact that there is no need to use the derivatization procedures. PMID- 9408855 TI - Determination of salbutamol enantiomers by high-performance capillary electrophoresis and its application to dissolution assays. AB - Capillary zone electrophoresis was successfully applied to the chiral separation of salbutamol after addition of a suitable cyclodextrin chiral selector to the electrophoresis buffer. Parameters important in achieving enantiomeric separation are cyclodextrin type, mobile phase pH and applied field strength. In our study, salbutamol enantiomeric separation was obtained with the following conditions: heptakis (2,6-di-O-methyl)-beta-cyclodextrin in 40 mM Tris (pH 2.5) and at 15 kV, obtaining a 3.09 resolution with migration times of 13.74 min for (R)-salbutamol and 13.98 min for (S)-salbutamol. Linearity, limit of quantitation, precision and accuracy were established using this method. The calibration curve was linear in a range of 1-40 micrograms ml-1 of racemic salbutamol (0.5-20 micrograms ml-1 of each enantiomer). This method was applied to evaluate the enantioselective release of salbutamol and taking into account the hypothesis that one enantiomer of a chiral drug would be released faster than the other from a pharmaceutical dosage form containing a racemic drug and a chiral excipient. For this purpose, matrix tablets formed by chiral excipients such as hydroxypropylmethylcellulose (HPMC) were considered. The release of the enantiomers of salbutamol from the formulations containing HPMC was found to be equivalent, with constant dissolution values (K) of 1.187 +/- 0.223% min-n for (R)-salbutamol and 1.076 +/- 0.268% min-n for (S)-salbutamol. PMID- 9408856 TI - Pharmacokinetic analysis of mizolastine in healthy young volunteers after single oral and intravenous doses: noncompartmental approach and compartmental modeling. AB - This paper presents the analysis of the kinetics of a new antihistamine, mizolastine, in 18 healthy volunteers, from concentrations measured after an intravenous infusion and two different oral administrations: tablet and capsule. Two approaches were used to analyze these data: (i) a noncompartmental approach implemented in PHARM-NCA; (ii) a compartmental modeling approach implemented in a new S-PLUS library, NLS2, which allows the estimation of variance parameters simultaneously with the kinetic parameters. For the compartmental modeling approach, two-compartment open models were used. According to the Akaike criterion, the best model describing the kinetics of mizolastine after oral administration was the zero-order absorption model. The kinetic parameters obtained with PHARM-NCA and NLS2 were similar. The estimated duration of absorption was greater for the tablets than for the capsules (with means equal to 1.13 hr and 0.84 hr respectively). After an intravenous infusion, the mean estimated clearance was 4.9 L/hr, the mean lambda 2-phase apparent volume of distribution was 89.6 L and the mean terminal half-life was 12.9 hr. PMID- 9408857 TI - Mixed effect modeling of sumatriptan pharmacokinetics during drug development. I: Interspecies allometric scaling. AB - Allometric scaling is an empirical examination of the relationships between the pharmacokinetic parameters and size (usually body weight), but it can also involve brain weight for metabolized drug. Through all species, the protein binding of sumatriptan is similar (14-16%), and its metabolic pathway undergoes extensive oxidative deamination involving the monoamine oxidase A isoenzyme. These similarities across species suggested the possible relevance of an allometric analysis. Toxicokinetic data were collected from rats, pregnant rabbits, and dogs in animal pharmacokinetic studies where sumatriptan was administered intravenously to the animals at doses of 5 mg/kg. 0.25 mg/kg, and 1 mg/kg, respectively. Animal data were pooled and analyzed in one step using a mixed effect modeling (population) approach. The kinetic parameters predicted in any species were close to the observed values by species: 77 L/hr vs. 80 L/hr in man for total clearance, 137 L vs. 119 L for distribution volume at steady state. The value of the mixed effect modeling approach compared to the two-step method was demonstrated especially with the possibility of including covariates to describe the status of animal (e.g., pregnancy) in the model. Knowledge of the animal kinetics, dynamics, and metabolism of a drug contributes to optimal and expeditious development. Valuable information for the design of the first-dose-in man study may emerge from more creative data analysis based on all the information collected during the preclinical and ongoing nonclinical evaluation of a new drug. PMID- 9408858 TI - A recirculatory model of the pulmonary uptake and pharmacokinetics of lidocaine based on analysis of arterial and mixed venous data from dogs. AB - Pulmonary uptake of basic amine xenobiotics such as lidocaine may influence the onset of drug effect and ameliorate toxicity. To date, pharmacokinetic analysis of pulmonary drug uptake has been only semiquantitative and ill-suited for relating pharmacodynamics to pharmacokinetics or for estimating the time course of the fraction of drug dose residing in the lung during a single pass. We have developed recirculatory models in an experiment in which lidocaine was injected into the right atrium simultaneously with markers of intravascular space (indocyanine green) and total body water (antipyrine); this was followed by rapid arterial and mixed venous blood sampling. Such models are interpretable physiologically and are capable of characterizing the kinetics of the pulmonary uptake of lidocaine in addition to peripheral tissue distribution and elimination. The apparent pulmonary tissue volume of lidocaine (39 ml/kg) was nearly ninefold greater than that of antipyrine (4.5 ml/kg). The recirculatory model characterized both arterial and mixed venous data, but the latter data were not essential for estimating lidocaine's pulmonary disposition either before or after recirculation of drug was evident. PMID- 9408859 TI - Pharmacokinetic-based minibolus delivery as an alternative to continuous infusion for drugs that exhibit a biophase lag. AB - The presence of a biophase compartment in a pharmacokinetic model indicates that the response to an administered dose of drug is damped such that the time to peak effect occurs after the peak concentration in the bloodstream. This phenomenon, which is common to most intravenous anesthetic agents, can be exploited by a drug delivery method that administers minibolus doses of drug rather than a continuous infusion. Through analysis of the frequency response behavior of the biophase compartment, a bolus magnitude and dose frequency or interval (1/frequency) can be chosen such that the oscillation in drug effect is minimized even though the plasma concentration may be changing significantly with each supplemental dose. A pharmacokinetic and pharmacodynamic based method for calculating the bolus dose size and dosing interval is presented. The trade-off between dose interval and change in drug effect is exemplified through computer simulation of this strategy applied to delivery of the neuromuscular blocking agent pancuronium. The method provides a repetitive perturbation to the pharmacokinetic and pharmacodynamic system that can aid in model parameter identification during closed loop applications. PMID- 9408860 TI - Bayesian population pharmacokinetic and pharmacodynamic analyses using mixture models. AB - Population studies of the pharmacokinetics or pharmacodynamics of drugs help us, learn about the variability in drug disposition and effects, information that can be used to treat future patients at safe and effective doses. We present a new approach to population modeling based on a weighted mixture of normal distributions having random weights and means. This method allows estimation of underlying continuous population distributions without prespecifying the parametric form or shape of these probability distributions. Additionally, this method can carry out nonparametric regression of pharmacokinetic or dynamic parameters on patient covariates while estimating the underlying distributions. Two examples illustrate the method and its flexibility. PMID- 9408861 TI - Bayesian nonparametric population models: formulation and comparison with likelihood approaches. AB - Population approaches to modeling pharmacokinetic and/or pharmacodynamic data attempt to separate the variability in observed data into within- and between individual components. This is most naturally achieved via a multistage model. At the first stage of the model the data of a particular individual is modeled with each individual having his own set of parameters. At the second stage these individual parameters are assumed to have arisen from some unknown population distribution which we shall denote F. The importance of the choice of second stage distribution has led to a number of flexible approaches to the modeling of F. A nonparametric maximum likelihood estimate of F was suggested by Mallet whereas Davidian and Gallant proposed a semiparametric maximum likelihood approach where the maximum likelihood estimate is obtained over a smooth class of distributions. Previous Bayesian work has concentrated largely on F being assigned to a parametric family, typically the normal or Student's t. We describe a Bayesian nonparametric approach using the Dirichlet process. We use Markov chain Monte Carlo simulation to implement the procedure. We discuss each procedure and compare our approach with those of Mallet and Davidian and Gallant, using simulated data for a pharmacodynamic dose-response model. PMID- 9408862 TI - Errors in clearance estimation after bolus injection and arterial sampling: nonexistence of a central compartment. AB - In conventional pharmacokinetics monotonic decreasing drug disposition curves are usually assumed after bolus injection. For arterial sampling starting about 1 min after dosing the resulting neglect of the initial concentration peak leads to a relative overestimation of clearance which may be approximated by the percentage of systemic drug extraction. This error can be avoided by administering the drug as a short-term infusion. PMID- 9408863 TI - Multiple pregnancy. New insights. PMID- 9408864 TI - Maternal mortality in twin gestations. AB - OBJECTIVE: To review various aspects of maternal morbidity and delivery complications leading to maternal death in twin gestations. STUDY DESIGN: Literature research. RESULTS: The incidence of maternal mortality associated with twin gestation is not known but can be estimated from a few community-based series and from studies on maternal morbidity related to twins. The combination of physiologic changes and perinatal pathologies that are either unique or more common in twin gestations (e.g., preeclampsia/eclampsia, need for tocolysis, anemia, abruption placentae, special delivery circumstances and cesarean section) is certainly increasing the maternal risk for serious morbidity and even mortality. CONCLUSION: Twin pregnancies should be included among the categories of pregnancy-related maternal death in order to learn how to eliminate avoidable mortality in twin gestations. PMID- 9408865 TI - Sex difference in crown-rump length in first-trimester twins. AB - OBJECTIVE: To determine if there is a difference in the crown-rump length (CRL) between male and female fetuses in the first trimester. STUDY DESIGN: CRL was determined in 77 normal twin pairs at the time of chorionic villus sampling. The sex of each fetus was assigned based upon the results of the sampling. RESULTS: A small difference was found between the CRL of males as compared to females, but this was not statistically significant. However, if there was a difference in size between the twin pairs, the odds were 1.5:1 that the larger would be male. CONCLUSION: There is no clinically significant difference in size between male and female fetuses in the first trimester of pregnancy. PMID- 9408866 TI - Terbutaline pump tocolysis in high-order multiple gestation. AB - OBJECTIVE: To review terbutaline pump tocolytic therapy as part of the management of high-order multiple gestations (triplet and quadruplet pregnancies). STUDY DESIGN: We performed a retrospective review of the medical records of triplet and quadruplet pregnancies cared for by Phoenix Perinatal Associates from August 1988 to January 1992 in whom terbutaline pump tocolysis was administered. RESULTS: The study group consisted of 15 triplet pregnancies and 6 quadruplet pregnancies. The 15 patients with triplets delivered at a mean (+/- SD) gestational age of 33.0 +/ 1.9 weeks. The six patients with quadruplets delivered at 33.0 +/- 1.3 weeks. Only 2 of 15 (13%) of the triplets and 1 of 6 (17%) of the quadruplets were delivered for tocolytic failure. CONCLUSION: Terbutaline pump tocolysis provides safe and effective tocolytic therapy in a select group of high-order multifetal gestations. PMID- 9408867 TI - Iatrogenic multiple pregnancy. Higher risk than a spontaneous one? AB - OBJECTIVE: To determine if spontaneous and induced multiple pregnancies have similar outcomes. STUDY DESIGN: We compared the results of antepartum and intrapartum surveillance and fetal outcome in spontaneous multiple gestations (group A) with induced multiple gestations (group B) at Albert Szent-Gyorgyi Medical University, Szeged, Hungary, in a six-year period. RESULTS: Between January 1, 1991, and December 31, 1996, there were 13,131 births; the number of multiple pregnancies was 307 (2.34%). There were 232 spontaneous and 48 induced twin pregnancies, 8 spontaneous and 16 induced triplet pregnancies, and 3 quadruplet pregnancies, all induced. Higher incidences of gestational diabetes and cervical insufficiency were found in group B. The incidences of prematurity in the induced and spontaneous groups were similar. The incidences of low birth weight and perinatal mortality were higher in induced triplet pregnancies than in spontaneous ones. Fetal outcome, with respect to Apgar score and umbilical cord blood pH, was much poorer in both induced groups. CONCLUSION: Iatrogenic multiple pregnancy following ovulation induction or assisted operative reproductive techniques may increase the incidence of pathologic events in the antepartum, intrapartum or postpartum period. Careful counseling before assisted reproductive techniques is of paramount importance. PMID- 9408868 TI - Zygosity testing. Current status and evolving issues. AB - This paper reviews the current status and evolving issues in the field of zygosity testing. Monozygotic twins are seldom absolutely identical. Traditional methods of zygosity testing, mathematical ones and assessment of physical characteristics are reviewed. The proper delineation of zygosity based on placentation and chorionicity is outlined. Genetic and environmental discordance patterns are described, with a discussion of why "identical" twins are never identical preceding specific guidelines for clinical practice. PMID- 9408869 TI - Twin-twin transfusion syndrome. Three possible pathophysiologic mechanisms. AB - OBJECTIVE: To propose a classification of twin-twin transfusion syndrome based upon three categories of placental anastomotic patterns. STUDY DESIGN: A mathematical model developed to compute fetal blood volume in monochorionic twins combines fetoplacental circulation with net fetofetal transfusion along placental anastomoses. We included (1) unequal cotyledonic sharing, assuming that smaller fractions cause smaller twins with lower blood pressure, and (2) significantly decreasing anastomotic resistance, combining Poiseuille's law with placental anastomotic growth. Fetoplacental compensatory mechanisms were not studied. RESULTS: First, unidirectional arteriovenous anastomoses produce steadily increasing fetal discordance by small anastomotic blood flow. Second, arteriovenous plus compensating anastomoses (venoarterial, arterioarterial, venovenous) produce fetal discordance followed by a dynamic steady state of minimal net fetofetal transfusion and large anastomotic flow. This circumstance mitigates further discordant growth. Third, unequal cotyledonic sharing plus superficial compensating anastomoses (arterioarterial, venovenous) produce fetal discordance followed by a steady state of equal fetal growth and small anastomotic flow. The model predictions include spontaneous disappearance and reversal of discordance. Serial measurement of fetal growth patterns and anastomotic flow could identify the syndrome's underlying pathophysiology. CONCLUSION: Testing the model predictions by relating clinical presentation with placental anatomy could increase our understanding and direct diagnostic and therapeutic strategies to match the underlying placental anatomy. PMID- 9408870 TI - Term and postterm twin gestations. Placental cause of perinatal mortality. AB - OBJECTIVE: To compare perinatal mortality in twins and singletons and to study the influence of fetal sex, placentation and maternal parity on perinatal mortality in term and postterm twin gestations. STUDY DESIGN: The subjects of the study were 1,511 twin pairs and 3,022 singletons. All were born at a gestational age of 27 weeks or more in two clinics in Amsterdam between 1931 and 1975. RESULTS: Perinatal mortality was lower in twins than in singletons until 37-38 weeks and higher afterwards. In twins, perinatal mortality was higher in boys than in girls, in monochorial than in dichorial twins, and in primiparae than in multiparae, especially in the last trimester. CONCLUSION: The development of the twin placenta may set limits in term and postterm twin gestations and may be responsible, to some extent, for the increase in perinatal mortality. PMID- 9408871 TI - Maternal position and ultrasonic cervical assessment in multiple pregnancy. Preliminary observations. AB - OBJECTIVE: To evaluate the influence of position on the functional anatomy of the cervix. STUDY DESIGN: Cervical length and width of the internal os were measured by transvaginal ultrasound in the recumbent and standing position from 15 gestational weeks to term in 15 twin and 3 triplet pregnancies. Measurements on asymptomatic primigravidae with twins who delivered vaginally after 36 weeks were made to calculate trends in normal twin pregnancies. These data were compared with that obtained in patients who had threatened preterm labor. RESULTS: As the total observation period progressed, the mean cervical length decreased from 50 to 27 mm in the recumbent position and from 48 to 21 mm in the standing position (P < .001). The differences between the values obtained in the recumbent and standing position increased from 5% to 31% as pregnancy progressed (P < .001). Funnelling was observed from 20 weeks onwards in the erect position but only after 35 weeks in the recumbent position. CONCLUSION: Our preliminary observations support a policy aimed at prevention of prematurity by reducing physical activity and standing work in patients who have multiple gestations. The full impact of these observations on the early detection of special risk groups and the prescription of preventive interventions have yet to be evaluated. PMID- 9408872 TI - Biparietal diameter in twins at gestational weeks 18-32. Differences and similarities. Le Groupe Romulus. AB - OBJECTIVE: To examine twin similarities on biparietal diameter (BPD) measurements by zygosity (monozygotic [MZ] and dizygotic [DZ]) and chorionicity (monochorionic [MC] and dichorionic [DC]) and their evolution during pregnancy. METHOD: A sample of 54 pairs of twins (43 DZ sets, 11 MZ sets [7 MC and 4 DC]) was constructed using retrospective data. Despite the small sample size, our data were complete, and, for the first time we measured different fetal parameters on digital ultrasound images outside routine examination. The intraexaminer and interexaminer reliability of BPD measurement was significant (r = .95, P < .001). RESULTS: In this study, developmental results indicate significant linear regression coefficient (R) through the whole period of gestation (r = .96, P < .001), though product moment correlations comparing the periods of gestation two by two are weaker. The distribution of BPD values was slightly wider at the 28th week and markedly wider at the 32nd week than that at the 18th and 23rd weeks. The intraclass correlations of DZ and MZ (MC and DC) twins were examined at the 18th, 23rd, 28th and 32nd weeks of amenorhea. The intraclass correlations of DZ twins were significant through the whole period of gestation (r = .45, P = .001; r = .27, P = .04; r = .36, P = .008; and r = .42, P = .002, respectively), whereas the intraclass correlations of MZ twins were significant only at the 18th, 23rd and 28th weeks (r = .73, P = .002; r = .69, P = .005; r = .49, P = .047, respectively). We found significant within-variance differences not only between DZ and DC-MZ but also between DC-MZ and MC-MZ in late gestation. CONCLUSION: Our analysis of twin BPD development demonstrated that zygosity and chorionicity type are both important determinants of twin fetal development. PMID- 9408873 TI - Nicotine and caffeine. Influence on prenatal hemodynamics and behavior in early twin pregnancy. AB - OBJECTIVE: To evaluate the effect of nicotine and caffeine in early multiple pregnancies. STUDY DESIGN: Ten women with twin pregnancies volunteered to take part in the study. Five mothers smoking > 5 cigarettes/d were investigated before and after smoking. Five women were investigated before and after drinking coffee. Maternal hemodynamics, umbilical blood flow, fetal heart rate and twin behavior were longitudinally documented during a 30-minute session from 8 to 16 gestational weeks. Fetal behavior was analyzed from videotapes using a one-minute window. RESULTS: The mean increase in maternal heart rate after smoking was 10 bpm (P < .005). After drinking coffee, it was 8 bpm (P < .01). The pulsatility index of the umbilical artery showed a significant increase after smoking (P < .01). After coffee the increase was also significant (P < .05). Two of five behavioral parameters (swallowing and breathing) demonstrated significant changes after the consumption of coffee (P < .001). CONCLUSION: This pilot study demonstrated significant effects of nicotine and caffeine consumption on maternal and fetal hemodynamics. It seems worthwhile to study the influences of smoking and coffee consumption on early twin behavior in larger samples. PMID- 9408874 TI - Epidemiologic paradox in multiple births among Asians in Illinois. Correlation between risk factors and outcomes. AB - OBJECTIVE: To determine if the plural birth rate, maternal risk factors and neonatal outcomes among Asian American populations residing in Illinois are homogeneous or heterogeneous with regard to maternal risk factors and neonatal outcomes and to attempt to establish correlations between maternal risk factors and neonatal outcomes. STUDY DESIGN: A total of 1,145,962 computerized birth certificate files were analyzed for 11 Asian subgroups giving birth in Illinois in the years 1989-1994. RESULTS: The multiple birth rates varied between 6 per 1,000 (Vietnamese) to 15 per 1,000 (Cambodian) (heterogeneity among all groups, P < .005). The percent of neonates born at gestational ages 22-33 weeks varied between 0 (Cambodian) to 28 (Thai). Japanese women showed the highest rate of neonates with birth weight less than 2,500 g after adjustment for race: 15.38 per 1,000. Women from Cambodia and Laos had the highest sum of maternal risk factors; those from the Philippines and Thailand had the least. The lowest collective adverse neonatal outcomes were in infants born to women from Cambodia and Laos. CONCLUSION: The 11 subgroups of Asians living in Illinois demonstrate heterogeneity with respect to multiple birth rate, selected maternal risk factors and adverse neonatal outcomes. After considering all maternal risk factors and neonatal outcomes, the two groups (Cambodians and Laotians) with the poorest maternal risk factors had the lowest rate of poor neonatal outcomes. This is the first time that this epidemiologic paradox has been observed in twins of Asian ancestry. PMID- 9408875 TI - Monoamniotic twin cord entanglement. A case report with color flow Doppler Ultrasonography for antenatal diagnosis. AB - BACKGROUND: Monoamniotic twinning is a rare event with a high perinatal mortality rate from cord accidents. To date, only a few cases have been reported in which cord entanglement was diagnosed with Doppler ultrasonography. CASE: In a monoamniotic twin pregnancy, umbilical cord entanglement was diagnosed at 20 weeks' gestation by color flow Doppler ultrasonography. The entanglement was associated with growth discordance between the twins and subsequent death of the smaller fetus. CONCLUSION: The rarity of this condition hinders our understanding of the pathophysiology and development of management protocols. PMID- 9408876 TI - High-order multiple pregnancies complicated by HELLP syndrome. A report of four cases with corticosteroid therapy to prolong gestation. AB - BACKGROUND: Previously, HELLP syndrome (hemolysis, elevated liver enzymes and low platelets) was perceived as an indication for immediate delivery. This often results in delivery of infants remote from term and with significant morbidity and mortality for both mother and neonates. While stabilization of maternal status has been reported to occur in singleton pregnancies with the use of antepartum administration of high-dose corticosteroids, it has not been reported to occur in high-order multiple pregnancies. We extended the use of corticosteroids to high-order multiple pregnancies complicated by HELLP syndrome to stabilize the disease process and lengthen gestation. CASES: Four cases of high-order multiple pregnancies complicated by HELLP syndrome were treated with high-dose, long-term corticosteroids to prolong gestation. The administration of dexamethasone resulted in stabilization of laboratory values and prolongation of gestation by 29, 6, 20 and 41 days, respectively. CONCLUSION: Antepartum administration of high-dose corticosteroids in high-order multiple pregnancies complicated by HELLP syndrome stabilized laboratory values and prolonged gestation in four patients remote from term. PMID- 9408877 TI - Selective feticide by embolization in twin-twin transfusion syndrome. A report of two cases. AB - BACKGROUND: The early development of complications in twin-twin transfusion syndrome is a poor prognostic sign. For this reason, new techniques for intrauterine therapy are being developed: invasive options, such as selective feticide of one of the twins, have been reported. CASES: Two cases of twin-twin transfusion syndrome in the late second trimester were treated by selective feticide using vascular embolization to the more severely damaged fetus. In one case the embolized fetus was a hydropic recipient with a normal cotwin; in the other, the donor fetus was affected by bilateral hydrocephalus. These fetuses underwent ultrasound-guided embolization using a bolus of histoacryl injected into the umbilical vein and fetal heart. Both patients went on to deliver healthy singletons in the third trimester. CONCLUSION: In twin-twin transfusion syndrome of early onset, embolization may help one of the twins survive. This technique is neither time-consuming nor expensive and does not require a general anesthetic. PMID- 9408878 TI - Vaginal lever pessary in patients with multiple gestation, preterm labor and low fetal station. A report of three cases. AB - BACKGROUND: The vaginal pessary has been utilized previously in patients with cervical incompetence, resulting in improvement in pregnancy outcome. The mechanical advantages generated by the vaginal lever pessary could theoretically be applied to patients in preterm labor with low station of the presenting part to prevent cervical dilatation. CASES: One patient with twins and two with triplet pregnancies presented in preterm labor with advanced cervical dilatation and low fetal station. They were treated with tocolytic drugs, and a vaginal lever pessary was placed. The gestational age at delivery was delayed with the addition of the vaginal pessary. Based on previous experience with these difficult cases, it was judged that the use of the pessary achieved a delay in delivery that would not have occurred without the device. CONCLUSION: Vaginal pessaries can be used as mechanical adjuvants in the treatment of preterm labor. These devices work by altering the pressure dynamics on the cervix and lower uterine segment. They may also help limit cervical change by preventing engagement of the presenting part into the maternal pelvis. In these cases, a vaginal pessary appears to be beneficial in delaying delivery in multiple gestations presenting with advanced cervical dilatation and low station of the presenting part. PMID- 9408879 TI - Complete hydatidiform mole and coexisting normal fetuses. A report of two cases with contrasting outcomes. AB - BACKGROUND: Multiple pregnancies consisting of a complete hydatidiform mole and coexisting fetuses are relatively rare but may become more common due to the increasing use of ovulation-induction agents. CASES: We report on a twin and a triplet pregnancy, conceived using clomiphene citrate, with contrasting outcomes. The twin pregnancy resulted in a term delivery of a healthy singleton and the triplet pregnancy in a termination at 17 weeks followed by the development of choriocarcinoma. CONCLUSION: The few cases available suggest that a subgroup of complete moles follows a more benign course and can be managed conservatively, allowing the pregnancy to go to term with appropriate follow-up, whereas other cases follow a more aggressive course. Larger case series are needed to develop definitive protocols. PMID- 9408880 TI - Immunotherapy for recurrent pregnancy loss: "standard of care or buyer beware". PMID- 9408881 TI - Ceramide stimulates prostaglandin production by human amnion and decidual cells. AB - OBJECTIVE: To determine whether ceramide regulates prostaglandin (PG) production by cultured human amnion cells and decidual cells independently of interleukin-1 beta (IL-1 beta). METHODS: Cells were grown in monolayer culture and then incubated with varying concentrations of ceramide, IL-1 beta, ceramide in the presence and absence of IL-1, and control media. Production of PGE2 was determined using a specific radioimmunoassay. RESULTS: Ceramide induced a significant concentration-dependent increase in PGE2 production by amnion cells and decidual cells. However, PGE2 production induced by IL-1 beta was significantly more than with ceramide alone, and there was no potentiation of PGE2 production with coincubation of ceramide and IL-1 beta. CONCLUSION: We suggest that term human amnion cells and decidual cells are responsive to ceramide independent of IL-1 beta and that generation of these substances in response to an infection in the uterus may lead to increased PG production by human gestational tissues, indicating that there are several mechanisms leading to PG production by these cells. PMID- 9408882 TI - Influence of estradiol and androstenedione on ACTH and cortisol secretion in the ovine fetus. AB - OBJECTIVE: To test the hypothesis that physiologic increases in fetal plasma 17 beta-estradiol and androstenedione modulate the activity of the fetal hypothalamic-pituitary-adrenal (HPA) axis. METHODS: Seventeen pregnant ewes and their fetuses were chronically catheterized. At the time of surgery, the fetuses received implants that released 17 beta-estradiol (n = 5) alone or 17 beta estradiol and androstenedione (n = 6), each at a rate of approximately 250 micrograms/day for each steroid. The control group (n = 6) received either no pellet (n = 2) or a "placebo" pellet, which contained no steroid (n = 4). Fetal blood samples were drawn for hormone and blood gas analysis at 1-3-day intervals until the time of spontaneous parturition. Fetal plasma ACTH and cortisol concentrations were fit to semilogarithmic equations and analyzed by stepwise multiple linear regression analysis for statistically significant effects of 17 beta-estradiol and androstenedione. RESULTS: Estradiol significantly increased and androstenedione significantly decreased the ACTH and cortisol concentrations. Treatment with both 17 beta-estradiol and androstenedione resulted in parturition approximately 4 days earlier than in the other groups (P < .05). CONCLUSIONS: Physiologic increases in fetal plasma estradiol and androstenedione modify the activity of the HPA axis. PMID- 9408884 TI - Flow decreases myogenic reactivity of mesenteric arteries from pregnant rats. AB - OBJECTIVE: To determine whether pregnancy alters the response of small mesenteric arteries to increased pressure (myogenic reactivity) and flow. METHODS: Mesenteric arteries (300 microns) from cycling nonpregnant (NP, n = 6) and late pregnant (20 days, LP, n = 6) Sprague-Dawley rats were dissected and mounted on an arteriograph system designed for the precise measurement of pressure and flow. Myogenic reactivity was measured as the percentage constriction after a pressure increase to 75 mmHg in the absence and presence of flow (60 microL/minute). RESULTS: In the absence of flow, there was no difference in myogenic reactivity in arteries from NP versus LP animals (NP, 8.4 +/- 1.4%; LP, 11.0 +/- 1.6%; not significant). In the presence of flow, myogenic reactivity was decreased in arteries from LP rats, but was unchanged in arteries from NP rats (NP, 13.2 +/- 1.1%; LP, 2.5 +/- 2.9%; P < .05). The differential group effect appeared to result not from differences in arterial response to changes in pressure or flow alone, but rather from the interaction between pressure and flow. CONCLUSION: These results suggest that pregnancy alters the interaction of the physical forces of pressure and flow on the arterial wall in a manner consistent with decreased vascular resistance. PMID- 9408883 TI - Pregnancy induces an increase in the expression of glyceraldehyde-3-phosphate dehydrogenase in uterine artery endothelial cells. AB - OBJECTIVE: We determined the effects of pregnancy on glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression in ovine uterine and omental (systemic control) artery endothelial cells (UAEC, OAEC). We also determined in primary cultures of UAEC the effects of exposure to either exogenous nitric oxide (NO) or angiogenic growth factors (basic fibroblast growth factor [bFGF], vascular endothelial growth factor [VEGF], and epidermal growth factor [EGF]) on UAEC GAPDH expression. METHODS: We isolated UAEC to high purity from both nonpregnant (NP; n = 4) and pregnant ewes (P; 110-120 days' gestation, n = 4) by limited collagenase dispersion and immediately extracted total RNA. In additional experiments performed in vitro, ovine UAEC isolated from NP ewes and maintained in culture (n = 3) were exposed to 1) 100 mumol/L sodium nitro-prusside for 0, 6, 12, or 24 hours or 2) 10 ng/mL bFGF, VEGF, EGF, or vehicle for 24 hours. Total RNA was then immediately extracted. A partial ovine GAPDH (oGAPDH) cDNA was isolated by reverse transcriptase polymerase chain reaction (RT/PCR) and sequenced. A one tube semiquantitative RT/PCR amplification assay was established, and GAPDH mRNA was subsequently quantified in all samples of total RNA. The PCR products were separated by size, quantified by Southern hybridization analysis, and normalized to 28S rRNA content. Expression of GAPDH protein was also measured by Western analysis of endothelium-derived protein from omental (n = 14) and uterine (n = 16) arteries from NP and P ewes. RESULTS: Pregnancy was associated with a 4.5 fold increase in GAPDH mRNA levels in UAEC, although in vitro exposure of primary cultures of UAEC from NP ewes to NO or angiogenic growth factors did not significantly change GAPDH mRNA expression. A 1.6-fold increase in GAPDH protein was detected in the uterine artery endothelium of P versus NP ewes, but no corresponding increase was found in omental artery endothelium. CONCLUSION: Pregnancy increases the expression of both GAPDH mRNA and protein in UAEC. Furthermore, the pregnancy-induced increase in GAPDH protein in the endothelium of the uterine artery appears specific, as it is not observed in the omental (systemic) artery. This induction is not, however, reproduced in vitro with exogenous NO or angiogenic growth factor treatment (up to 24 hours). PMID- 9408885 TI - Induction of immune responses to ovarian tumor antigens by multiparity. AB - OBJECTIVE: Because epidemiologic data indicate a reduction in ovarian cancer risk with increased parity, the occurrence of maternal immunization against ovarian tumor-associated antigens during pregnancy was investigated. METHODS: Sera were obtained from nulligravid and multiparous women and from men. Cellular proteins were isolated from four ovarian tumor cell lines as well as from normal ovaries. These proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the presence of cellular proteins reactive with each individual's serum was assessed by Western immunoblot. Tumor-reactive antibodies from two multiparous women were used to prepare immunoaffinity columns for the isolation of reactive proteins from ovarian tumor cells. These immunoaffinity purified antigens were transferred electrophoretically to nitrocellulose membranes, stained with Ponceau S, and identified by amino acid sequencing. RESULTS: Western immunoblot analysis of the cellular proteins from four established ovarian tumor cell lines using sera from multiparous women as the primary antibody indicated that these samples recognized multiple bands on ovarian tumors, ranging from 30 to 150 kD. Two commonly recognized proteins were isolated and subjected to microsequencing, which identified the 56-kD band protein as elongation factor-1 alpha and the 38-kD protein as nucleophosmin/B23 protein. Both of these proteins play integral roles in cell growth. CONCLUSION: These findings suggest that certain antigens expressed by the fetus immunize women during pregnancy. This immune response may protect these women from the subsequent development of cancer. PMID- 9408886 TI - Expression of 11 beta-hydroxylase (CYP11B1) and aldosterone synthase (CYP11B2) in the human fetal adrenal. AB - OBJECTIVE: To understand better the steroidogenic capacity of the human fetal adrenal (HFA), we evaluated the expression of 11 beta-hydroxylase (CYP11B1) and aldosterone synthase (CYP11B2) in the fetal zone and neocortex of the HFA using a specific RNase protection assay. METHODS: Adrenal glands were obtained at the time of elective termination of pregnancy. Whole adrenals (n = 7) were frozen in liquid nitrogen, and subsequently total RNA extraction was performed by tissue homogenization followed by guanidinium/chloroform purification. In addition, RNA was obtained from separated fetal zone (n = 4) and neocortex (n = 4) tissues obtained by dissection. RNase protection assays were then performed using radiolabeled complementary RNA probes generated by T7 RNA polymerase directed against transcripts for CYP11B1, CYP11B2, and actin, the latter of which was used as a control for RNA integrity. Transcripts also were examined using a reverse transcription polymerase chain reaction (RT-PCR) protocol specific for CYP11B1 or CYP11B2. RESULTS: The RNase protection assay was designed to distinguish specific bands that corresponded to CYP11B1 (232 bp), CYP11B2 (262 bp), and actin (221 bp). RNA isolated from whole HFA was observed to have high levels of CYP11B1 transcript, whereas CYP11B2 was not detected. Dissected neocortex and fetal zones were found to contain transcript for CYP11B1 using both the RNase protection assay and RT-PCR analysis. In contrast, using the RNase protection assay, CYP11B2 mRNA was not observed in the RNA from the fetal zone, but after prolonged exposure there was a band corresponding in size to CYP11B2 observed in RNA from the neocortex. Using the more sensitive RT-PCR method, transcript for CYP11B2 was found in both neocortex and fetal zone. CONCLUSION: The HFA expresses low levels of CYP11B2 in accordance with its low production of mineralocorticoid. The expression of CYP11B1 in the fetal zone is intriguing because this enzyme is not necessary for the production of C19 steroids. Definition of the molecular mechanisms controlling expression of the CYP11B genes will be necessary to determine why the HFA differentially expresses these isoenzymes. PMID- 9408887 TI - Regulation of PTP1D mRNA by peptide growth factors in the human endometrial cell line HEC-1-A. AB - OBJECTIVE: To assess, in the human endometrial cell line HEC-1-A, the presence of protein tyrosine phosphatase 1D (PTP1D) and the possible regulation of its mRNA expression by mitogens such as forskolin (an agent that increases intracellular cyclic adenosine monophosphate [cAMP] levels), epidermal growth factor (EGF), and insulin-like growth factor-I (IGF-I). METHODS: Cells were grown to confluence and maintained in serum-free media for 24 hours before treatment. Cells were exposed to forskolin, EGF, and IGF-I for increasing time periods (0, 1, 3, 6, and 24 hours), and PTP1D mRNA expression was determined by Northern blot analysis. In addition, cells were incubated with increasing doses of forskolin (final concentrations: 1, 5, 10, 20, and 30 mumol/L) for 6 hours. RESULTS: When treated with the various mitogens, cells increased their stimulation of PTP1D mRNA expression in a time- and dose-dependent fashion. Specifically, forskolin, EGF, and IGF-I induced maximal mRNA expression at 6, 3, and 6 hours, respectively. Expression induced by forskolin, EGF, and IGF-I was five, three, and six times control levels, respectively. At a dose of 10 mumol/L, forskolin induced PTP1D mRNA expression almost two times higher than control values. CONCLUSION: These data suggest that in human endometrial carcinomas, cAMP, EGF, and IGF-I may regulate the expression of PTP1D mRNA, which may, in turn, play a role in uncontrolled cell proliferation and neoplastic transformation. PMID- 9408888 TI - Human umbilical vessels and cultured umbilical vein endothelial and smooth muscle cells lack detectable protein and mRNA encoding estrogen receptors. AB - OBJECTIVE: To identify and characterize estrogen receptors in human umbilical vascular tissues and in cultured cells derived from the human umbilical vein. METHODS: Human umbilical vein endothelial (HUVE) and human umbilical vein smooth muscle (HUVSM) cells were isolated. Immunohistochemical, radioligand binding. Western immunoblotting, and reverse transcription-polymerase chain reaction (RT PCR) methods were used to detect estrogen receptors in vascular tissues and in cells derived from the umbilical cord. RESULTS: Estrogen receptor protein was not detected in either umbilical vessel tissue or in isolated HUVE or HUVSM cells. Messenger RNAs for the classic estrogen receptor (alpha) and estrogen receptor beta isoforms also were undetectable by RT-PCR. CONCLUSION: These findings suggest that the effects of estradiol observed in this widely used vascular model are mediated by very low concentrations of receptors that evade standard methods of detection. Alternatively, this steroid may affect umbilical vascular cells through mechanisms that do not involve the classic genomic estrogen-receptor pathway. PMID- 9408889 TI - Malignant lymphoma of the skin and superficial lymph nodes in a bull mastiff bitch. PMID- 9408890 TI - Acute effects of an anionic diet on bone mineral homeostasis in the bovine. AB - Fifteen Friesian oxen between 12 and 18 months of age with a mean body mass of 240.7 kg were randomly assigned to diets containing 0.25% phosphorus (P) or less to evaluate the acute effects of an acidiogenic diet of -11.1 meq/100 g of diet dry matter, compared with a basiogenic diet of +25.6 meq/100 g or a control diet of +16.5 meq/100 g of diet dry matter calculated as (Na + K)-(Cl + S), on blood, bone and faecal P, calcium (Ca) and magnesium (Mg) for a period of 9 weeks. Blood, bone and faecal responses to an anionic diet are described. An inverse relationship existed between bone and blood Ca, in which there was resorption from bone with increased blood Ca in response to the anionic diet. The anionic treatment group demonstrated simultaneous increases in bone, blood and faecal P concentrations at various stages of the experiment compared to the cationic and control treatment groups. Results indicate independent absorption and resorption of Ca and P into and out of bone. There was wide variation in the bone Ca:P ratio between 2.02 and 1.51 among animals fed the anionic diet, with the Ca:P ratio following Ca values and not bone P values. Bone and blood P had a linear relationship with dietary cation:anion balance (DCAB), increasing as the diet became more anionic in nature, but faecal P was curvilinear with highest concentrations at -11.1 and +25.6 meq/100 g compared to +16.5 meq/100 g. Concurrent blood, bone and faecal P increases at some stages of the experiment indicate a P-sparing effect of the anionic diet and warrants further research into the long-term effects of anions in the diet, leading to their use as a possible addition to improved licks in P-deficient areas. PMID- 9408891 TI - The effect of dietary protein on reproduction in the mare. I. The composition and evaluation of the digestibility of dietary protein from different sources. AB - Four rations that differed in their crude protein and essential amino-acid content were compiled. Digestibility of the crude protein and essential amino acid contents were determined biologically in a feeding trial using 4 Anglo-Arab stallions. Their respective daily diets were: Diet 1:2 kg cubes, 5 kg tef hay (Eragrostis tef); Diet 2:2 kg cubes, 5 kg lucerne hay (Medicago sativa); Diet 3:2 kg cubes, 5 kg tef hay, 200 g fishmeal; Diet 4:2 kg cubes, 5 kg lucerne hay, 200 g fishmeal. The concentrations of the amino-acids threonine, iso-leucine, leucine and arginine were increased in the total ration when lucerne hay replaced the tef hay while fishmeal supplementation increased the methionine and lysine contents, which provided a wide-range of concentrations of digestible amino-acids in each of the 4 rations. PMID- 9408892 TI - The effect of dietary protein on reproduction in the mare. II. Growth of foals, body mass of mares and serum protein concentration of mares during the anovulatory, transitional and pregnant periods. AB - The effect of 4 different diets, in terms of protein quantity and quality, on total serum protein (TSP), albumin and globulin was investigated. Non-pregnant mares that were not lactating (n = 36), pregnant mares that had foaled (n = 24) and their foals (n = 24) were used in this study. Daily total protein intake had no effect on blood protein concentrations in the mares. Total protein intake and quality (available essential amino-acids) did affect the body mass of mares during lactation. When mares were fed the minimum recommended (National Research Council 1989) total daily protein, foal mass decreased by approximately 25% at weaning compared to the foals whose dams were on a higher level of protein intake. The TSP concentrations of foals at birth were on average 10 g/l lower than those of the mares. Albumin concentrations of foals during the first 60 days of life were on average 2-3 g/l lower than those of the mares. Globulin concentrations of foals were approximately 5 g/l lower than those of mares at weaning. PMID- 9408893 TI - The effect of dietary protein on reproduction in the mare. III. Ovarian and uterine changes during the anovulatory, transitional and ovulatory periods in the non-pregnant mare. AB - In the main experiment the total daily protein intake and quality (essential amino-acids) was varied in 4 groups of mares. The incidence of oestrus in mares during the transitional period was unaffected by protein nutrition. Ovarian activity, as evaluated by follicular development and size of the ovaries, was affected. Mares that received low-quality protein (Groups 1 and 2) had a higher number of smaller follicles (< 10 mm) that developed during the transitional period compared to mares on a high-quality protein intake (Groups 3 and 4). The mares that received the high quality protein ovulated 2-3 weeks earlier in the breeding season in a synchronised period of 4-5 weeks compared to a period of 6-8 weeks in Groups 1 and 2. The duration of the subsequent oestrous cycles was not affected. There was no difference in the diameter of the largest follicle of mares between groups on the day before ovulation. In a separate experiment, 5 maiden Anglo-Arab mares, 4-5 years of age, were slaughtered at different stages during the anovulatory, transitional and ovulatory periods of the breeding cycle. The morphology of the ovaries and uteri of these mares was described and photographed for use as guidelines when comparing ovarian changes and follicular activity of mares. PMID- 9408894 TI - Comparison of electron microscopy, enzyme-linked immunosorbent assay and latex agglutination for the detection of bovine rotavirus in faeces. AB - The performance characteristics of 2 enzyme immunoassays (ELISAs) and 4 latex agglutination assays (LXs) were evaluated for the detection of bovine rotavirus in faecal specimens of young calves with diarrhoea. A total of 26 specimens from calves less than 5 months of age were examined with different commercial assays and compared with electron microscopy (EM) as the gold standard and with polyacrylamide gel electrophoresis (PAGE) for the detection of atypical, non group A rotaviruses. In the 2nd study, EIA (Dako) and LX (Murex), the assays of choice, were used to analyse 97 further faecal specimens from calves with diarrhoea. The ELISAs proved to be the most sensitive compared with the other tests used. The EM and PAGE are 100% specific although slightly less sensitive than the commercial assays. The results show that all the commercial assays can accurately detect rotavirus in the stools of calves with gastroenteritis, although the suitability and choice of assay will depend upon the requirements of individual laboratories. PMID- 9408895 TI - Vermeersiekte caused by Geigeria burkei Harv. subsp. burkei var. hirtella merxm. in the northern province of South Africa. AB - The 1st field outbreak of vermeersiekte induced by Geigeria burkei Harv. subsp. burkei var. hirtella Merxm, is reported. It is also the first recorded outbreak of this disease in the arid sweet bushveld of the Northern Province of South Africa. The toxicosis was experimentally reproduced in a sheep following daily intraruminal administration of 2.5-5.0 g/kg dried, milled plant material for 18 consecutive days. Neither the sheep in the field outbreak nor the ewe in the experiment exhibited any signs of regurgitation of rumen contents (vermeersiekte). All developed only the stiff or paretic/paralytic forms of the disease. Serum activities of CK and GGT were slightly raised in clinically affected sheep (n = 11) during the field outbreak, and serum activities of AST, GLDH, GGT, LDH and CK increased in the ewe dosed with the plant material. Analysis of dried, milled Geigeria plant material confirms that this species is moderately nutritious. PMID- 9408896 TI - Standing laparoscopically-aided ovariectomy in mares. AB - Bilateral ovariectomy was performed in 11 mares and unilateral ovariectomy in 2 mares. The horses were standing and sedated for surgery. After appropriate preparation a laparoscope was inserted into the abdomen through the paralumbar fossa and the ovary was identified and anaesthetised with local anaesthetic via a custom-built needle. The ovary was then withdrawn from the abdomen through a separate flank incision and removed. The abdomen was not distended with gas before surgery. This method proved to be minimally invasive, rapid and effective. PMID- 9408897 TI - Clinical and epizootiological study of a leptospirosis outbreak due to Leptospira canicola in a feedlot. AB - This report describes the epizootiology, clinical presentation, diagnosis and treatment of an outbreak of leptospirosis caused by Leptospira canicola in feedlot calves. The infection appeared to be of high morbidity with a cumulative clinical incidence of 15.6%, cumulative subclinical incidence of 39% and high mortality (8.3%). Clinical disease was diagnosed in 4-8-month-old calves, while subclinical infection occurred in 9-12-month-old calves. Subclinical infection was based on serological evidence only. The zoonotic aspects of the infection are emphasised. PMID- 9408898 TI - Urban low-income African American men, HIV/AIDS, and gender identity. AB - In a 1993 Human Organization article, Jerome Wright called for more research on African American male sexual behavior and the risk for HIV infection. The present article is a response to that call. Wright pointed out a well-known fact of HIV/AIDS prevention programs: such programs have not been very successful in reaching low-income African American males. The present article suggests that perhaps the key to better understanding sex-related health-risk behavior is to conduct more systematic research on gender identity, and the historical and sociocultural origins of such identities. I argue that if we are truly interested in developing effective HIV/AIDS programs targeting low-income African American males, then the sociocultural "meanings" that this population attaches to AIDS related phenomena must be understood in the broader contexts of American constructs of masculinity, and in the real and perceived experiences of black men in America. Data from several ethnographic and qualitative research projects carried out among low-income African American male and female residents of Baltimore, other parts of Maryland, and Washington, D.C. are used in support of my primary arguments. PMID- 9408899 TI - Gender, sexuality, and health in a turn-of-the-century "black metropolis". PMID- 9408900 TI - African American males and HIV: the challenge of the AIDS epidemic. PMID- 9408901 TI - Signifying the pandemics: metaphors of AIDS, cancer, and heart disease. AB - This article offers a symbolic analysis of the cultural construction and signification of three of the major "pandemics" of the late 20th century: AIDS, cancer, and heart disease. It is based on unstructured interviews conducted in Israel between 1993-94 with 75 nurses and 40 physicians and between 1993-95 with 60 university students. Two key symbols, "pollution" and "transformation," are shown to constitute AIDS and cancer within a symbolic space that I suggest is "beyond culture," where body boundaries are dissolved and cultural categories are dismantled. Heart disease, in contrast, is metaphorized as a defect in the "body machinery." The article concludes by arguing that heart attack is depicted as the pathology of the Fordist, modernist body, while AIDS/cancer are pathologies of the postmodern body in late capitalism. PMID- 9408902 TI - Conjuring medical science: the 1986 referendum on AIDS/HIV policy in California. AB - Medical science occupies a peculiar status in American life. On the one hand, people often view medical science as a privileged and authoritative body of knowledge that transcends other kinds of knowledge. On the other hand, medical scientific authority can be easily conjured from the popular symbols of science, e.g., credentials, technical terms, and white lab coats. This problem can be converted into an anthropological question of meanings and symbols, based on Geertz's interpretive anthropology and Baudrillard's sociology of hyperreality. This article uses these frameworks to explore the cultural construction of medical-scientific authority in the case of a 1986 referendum on AIDS/HIV policy in California. The interpretation of that construction raises some difficult problems concerning anthropology's treatment of medical science. PMID- 9408903 TI - "You have to be your own doctor": sociocultural influences on alternative therapy use among gay men with AIDS in west Hollywood. AB - This article considers the use of alternative therapies by gay men with AIDS in West Hollywood, California. In all, 89 different alternative therapies for HIV and AIDS treatment were uncovered during the course of the study. In addition to the high number of alternative therapies, a majority of the men in the study (69.2 percent) were currently using some type of alternative treatment. Sociocultural influences on the use of alternative therapies are explored, including AIDS activism. Because most of the men (92 percent) also use Western medicine as well, the interface of these two types of treatments is also explored. PMID- 9408904 TI - Speed, sex, gay men, and HIV: ecological and community perspectives. AB - Fifteen years into the HIV/AIDS epidemic, a great deal is now known about the different populations impacted by the disease, including those affected directly or indirectly by drug use. Anthropology has played a critical role in assisting with this task by identifying hidden populations, developing new methodological approaches, and targeting outreach efforts. In spite of this considerable body of ethnographic knowledge, men who have sex with other men (i.e., MSM, or gay and bisexual men) who use drugs have not received the same research attention as other drug users, despite the fact that they represent nearly one-fifth of AIDS cases in the U.S. with injection drug histories. In response to the alarming increase in HIV seroprevalence among this population, this ethnographic project provides preliminary data about those who are at dual risk for HIV through both homosexual behavior and injection drug use. PMID- 9408905 TI - Hemopoiesis in human fetal and embryonic liver. AB - In this review, we describe the topographic distribution of hemopoietic cells of the lymphoid, myeloid, and erythroid lineages in the human fetal and embryonic liver. The data are based on studies of frozen tissue, allowing the determination of a broad range of hemopoietic antigens, and studies on paraffin-embedded tissue, allowing the combination of optimal morphology and immunodetection of lineage-specific antigens. The different hemopoietic lineages each show their own immunophenotype and distribution; intercellular and microenvironmental relationships were easily determined. In a few cases, some scarce CD34-positive early progenitor cells were seen. The number of proliferating cells, identified by monoclonal antibody (MAb) Ki-67, varied from 350 to 2500 (median = 1,500) per square millimeter of tissue. Erythroid cells reacted with antisera to glycophorin A, CDw75, and CD43 and partly surround a central macrophage, whereas the myelomonocytic cells reacted with CD45, CD43, CD74, and antilysozyme serum, and with LN3 from 14 weeks onward. Myelopoietic (CD15 positive) cells were localized mainly around portal triad vessels and increased in number with gestational age. The lymphoid cells showed CD45, CD43, CD45RA/MT2, CD45RA/MB1, MB2, and CD74 reactivity. B cells and their precursors were scattered among the hepatocytes without any sign of focal development in the age range studied. We seldom found cells positive for delta-H chain or C3bR (CD35); C3dR (CD21)-positive cells were even more scarce. Cells reactive with MAb WT1 (CD7) were present in a scattered single pattern (< or = 20/mm2) among the parenchymal cells; cells expressing mature T-cell markers (CD2, CD3, CD5) were rare. Large (> 15 mu) CD43-positive hemopoietic cells in the fetal liver were distinguished that exclusively expressed CD43, probably representing early hemopoietic progenitor cells. PMID- 9408906 TI - Origin and fate of the central macrophages of erythroblastic islands in the fetal and neonatal mouse liver. AB - Erythroblastic islands were examined by ultrastructure and ultrastructural histochemistry in fetal and early neonatal livers of the mouse. The liver primordium of day 11 embryos contained not only immature hemopoietic cells in the hepatic cords but also macrophages in expanded sinusoids. At 12 and 13 days of gestation, macrophages bearing large cytoplasmic inclusions increased in number, and some of them moved from the sinusoids into the hepatic cords. A ring of erythroblasts surrounded the macrophages, and erythroblastic islands could be identified at 14 days of gestation. Fetal livers contained two kinds of macrophages: sinusoidal macrophages and central macrophages of the erythroblastic islands. These macrophages exhibited a similar binding pattern to Griffonia simplicifolia isoagglutinin-IB4 (GS-IB4) and soybean agglutinin (SBA). Fetal hepatocytes, however, did not appear to engage in active phagocytosis, and the binding patterns of GS-IB4 and SBA differed significantly between hepatocytes and the two kinds of macrophages. In the early postnatal mouse, a marked decrease in the number of erythroblastic islands occurred. Erythroblasts left the central macrophages, and the macrophages subsequently underwent degeneration. The erythroblastic islands finally disappeared at the end of liver hemopoiesis, and the degenerated central macrophages were removed by scavenger macrophages in the perisinusoidal space. Our data demonstrate that scavenger macrophages play an essential role in the development of hepatic hemopoiesis, with special reference to the formation and dissolution of erythroblastic islands. PMID- 9408907 TI - Development and maintenance of bile canaliculi in vitro and in vivo. AB - The apical surfaces of hepatocytes are specialized to form the boundaries of the bile canaliculi. The canaliculi function to secrete and concentrate components of the bile and to transport the bile out of the interior of the hepatic parenchymal tissue to the epithelium-lined bile ducts. Failure of the canaliculi to form and function properly can lead to biliary stasis or release of bile components into the bloodstream, both potentially life-threatening situations. Experimental analysis of canaliculus development and function has been undertaken in a number of experimental systems, ranging in complexity from intact animals to isolated hepatocyte cell cultures. These approaches each have inherent advantages and disadvantages for studying the various aspects of canaliculus development and function. This article summarizes what is known about how the functional components of the canaliculus develop and the directions that current experimental approaches are leading in analyzing this process. Studies of model epithelial systems have begun to define how interactions between components of the cytoskeleton and plasma membrane regulate the structure of polarized plasma membranes. These results are also discussed in terms of the bile canaliculus. PMID- 9408908 TI - Developmental appearance of ammonia-metabolizing enzymes in prenatal murine liver. AB - To resolve an apparent discrepancy in the developmental appearance of glutamine synthetase (GS) protein in rat [Gaasbeek Janzen et al. (1987) J. Histochem, Cytochem., 35:49-54] and mouse [Bennett et al. (1987) J. Cell Biol., 105:1073 1085] liver, we have investigated its expression during liver development in the mouse and compared it with that of carbamoylphosphate synthetase I (CPS). The expression of glutamate dehydrogenase was used as a marker to identify all hepatocytes in these strongly hematopoietic livers. GS protein accumulation starts in mouse hepatocytes at embryonic day (ED) 15. The first hepatocytes in which the enzyme accumulates were found around the major hepatic veins. CPS protein was found to accumulate in mouse hepatocytes from ED 13 onward: first, at the center of the median and lateral lobes, but temporarily not at the periphery of these lobes and not at the caudate lobe. The initial phase of accumulation of GS and CPS protein was characterized by a heterogeneity in enzyme content between hepatocytes. By ED 17, both enzymes were detectable in all hepatocytes at the center of the median and lateral lobes. This event marked the onset of the development of the complementary distribution of the enzymes typical of zonal heterogeneity in the adult mammalian liver. However, during the perinatal period, the pericentral hepatocytes temporarily accumulated CPS protein. We also observed heterochrony between species in the appearance of CPS protein in the small intestine. PMID- 9408909 TI - Expression and inducibility of P450 enzymes during liver ontogeny. AB - Several approaches, including immunoquantification of individual enzymes, profiling of substrate activities by immunoinhibition using highly specific polyclonal and monoclonal antibodies, and the estimation of corresponding mRNAs with nucleic acid probes, have been used to investigate the ontogeny of cytochromes P450 in livers of rodents and man. CYP1A1 is expressed very early in development in rodents, whereas most other enzymes either appear at or near birth (CYP2B, CYP2C23, and CYP3A) or between 2 and 4 weeks following birth (CYP2A, CYP2C6, CYP2C7, CYP2C11, CYP2C12, and CYP4A10). The constitutive expression of enzymes is subject to regulation by various transcriptional nuclear and/or hormonal factors (CYP2B and CYP2C) or in a sex-dependent manner (CYP2A, CYP2C11, CYP2C12, CYP3A, and CYP4A10). The enhanced sensitivity and specificity of immunocytochemical and in situ hybridisation studies have revealed differences, with age and xenobiotic treatment, in the intercellular expression of certain P450 enzymes of the liver. For example, in rats, the expression of CYP1A1 and 1A2 is differentially regulated at the level of the individual cell from as early as 24 hours before birth. The human foetal liver relative to rodents has a substantial level of CYP3A and also has the capacity to metabolise a greater repertoire of substrates. Evidence to date suggests that P450 enzymes in man are regulated in a manner similar to that in other animals. The balance between different individual enzymes of cytochrome P450 in foetuses and/or neonates is subject to modulation by xenobiotics, the consequences of which may lead to toxicologically compromised livers with respect to metabolic handling of certain substrates. PMID- 9408910 TI - Spatial distribution of cytoskeleton intermediate filaments during fetal rat hepatocyte differentiation. AB - The construction of the liver parenchyma throughout fetal development depends on the elaboration of intercellular contacts between epithelial cells and between epithelial and mesenchymal cells. During this time, the spatial distribution of cytokeratins in hepatocytes shows a striking evolution as demonstrated by confocal microscopy and image analysis. In the early stages of fetal rat development, the liver is mainly a hematopoietic organ and hepatocytes represent fewer than 40% of all liver cells. At this time, cytokeratin filaments are scarce and are randomly distributed inside the cytoplasm. A coexpression of desmin and cytokeratin is found in some cells. Intercellular contacts between epithelial and mesenchymal cells are more numerous than between epithelial cells. Later in development, hepatocytes are arranged in a "muralium duplex" architecture (two cell-thick sheets). Contacts between hepatocytes become more numerous and bile canaliculi become well developed. The density of cytokeratin filaments increases and appears to be very high near the bile canaliculi. In adult liver, hepatocytes are arranged in a "muralium simplex" architecture. Cytokeratin filaments show a symmetrical distribution in relation to the nuclear region. The highest density of filaments is found near the cytoplasmic membrane. Variations of the spatial distribution of intermediate filaments throughout hepatocyte differentiation were investigated in a pilot study using computerized image analysis. We found significant differences between the filament networks in fetal and adult hepatocytes. PMID- 9408911 TI - Biogenesis and function of lipolysosomes in developing chick hepatocytes. AB - Proliferation of lipolysosomes is one of the characteristic aspects of embryonic chick hepatocytes. Formation of lipolysosomes is observed in the well-developed trans-Golgi network, with the highest frequency occurring from 11 to 14 days of incubation. The lipolysosomes usually contain a small and electron-dense lipid inclusion; however, during development, they gradually enlarge with an accompanying reduction in the electron density of the inclusion. Lipolysosomes isolated from neonatal chick liver homogenates were mainly composed of esterified cholesterol and showed considerably high activity of lysosomal enzymes. Moreover, the lipolysosome fraction is clearly shown to be a function of intralysosomal lipolysis via acid lipase. This accumulation of esterified cholesterol within lipolysosomes might be attributed to an excessive uptake and conversion of plasma lipoproteins to lipolysosomes. This concept is supported by the appearance of an abundance of coated pits and both "early" and "late" endosomes. The major components of plasma lipoprotein are low density lipoprotein (LDL) and high density lipoprotein (HDL), the cholesterol-rich lipoproteins, whose cholesterol content increases during the last week of incubation when the lipolysosomes quickly enlarge. Plasma lipoprotein particles are produced in the yolk sac epithelium from yolk very low density lipoprotein (VLDL) and transferred via the vitelline circulation to the chick liver. After hatching, when the supply of nutrients from the yolk sac is terminated, lipolysosomes immediately decrease in size and number. The cholesterol and fatty acids released are useful as an energy source and lipid metabolism in general, especially after hatching. Food intake induces the use of and accelerates the disappearance of lipolysosomes. Instead of lipolysosomes, lipid droplets appear and increase in number and size with concomitant increases of triglyceride concentrations in the liver homogenates, suggesting that lipogenesis has begun in the chick hepatocyte. PMID- 9408912 TI - Biogenesis of peroxisomes in fetal liver. AB - Peroxisomes are single membrane-limited cell organelles that are involved in numerous metabolic functions. Peroxisomes do not contain DNA; the matrix and membrane proteins are encoded by the nuclear genome. It is assumed that new peroxisomes are formed by division of existing organelles. The present article gives an overview of microscopic studies and recent unpublished results dealing with peroxisome biogenesis in mammalian fetal liver and presents data on peroxisomes in oocytes. Cytochemical (catalase and D-aminoacid oxidase activity) and immunocytochemical data in rat and human liver (antigens of catalase, the three peroxisomal beta-oxidation enzymes, alanine: glyoxylate aminotransferase, peroxisomal membrane proteins with molecular weights of 42 and 70 kDa) indicate that during embryonic and fetal development the peroxisomal population undergoes a differentiation with respect to the composition of the matrix and to the size and number of the organelles. In the youngest stages, rare and small peroxisomes are present, into which the matrix components are imported in a sequential way. The import seems asynchronous in peroxisomes of the same hepatocyte. The size and number of the peroxisomes increase during liver development. In rat and human liver, no morphological or immunocytochemical evidence for an elaborate network of interconnected peroxisomes ("reticulum") was found. Instead, peroxisomes presented as individual organelles, which occasionally show membrane extensions. The importance of the metabolic functions of peroxisomes in human liver is emphasized by the peroxisomal disorders. In the liver of affected fetuses, the microscopic features associated with the defect can already be recognized; i.e., either catalase containing peroxisomes are absent and catalase is localized in the cytoplasm (in fetuses affected with Zellweger syndrome or with infantile Refsum disease) or peroxisomes are present but they are abnormally enlarged (e.g., a fetus affected with acyl-CoA oxidase deficiency). In the quail ovary, numerous peroxisomes are observed in the oocyte and in the granulosa cells during follicle maturation, but not in the full-grown egg. Thus, the mechanism of peroxisome inheritance remains unresolved. PMID- 9408913 TI - Reproducibility of proton magnetic resonance spectroscopic imaging in patients with schizophrenia. AB - Using proton magnetic resonance spectroscopic imaging (1H-MRSI) we found in a previous study a specific pattern of neuronal pathology in patients with schizophrenia as determined by relative loss of signal from N-acetyl-containing compounds (NAA). The purpose of the present study was to assess the reproducibility of the results of 1H-MRSI both in patients with schizophrenia and in normal controls. We studied twice 10 patients and 10 controls on 2 days separated by, on average, 3 months. Reproducibility was assessed with several statistical procedures including ANOVA, coefficients of variation (CVs) and intra class correlation coefficients (ICC). Patients showed significant reductions of NAA/creatine-phosphocreatine (CRE) and NAA/choline-containing compounds (CHO) selectively in the hippocampal region (HIPPO) and in the dorsolateral prefrontal cortex (DLPFC) on both experimental days. A repeated measures ANOVA showed no effect of time on metabolite ratios in all subjects. CVs were fairly low (especially for NAA/CRE and CHO/CRE) and did not differ significantly between patients and controls. The ICCs of the ROIs reached statistical significance only in a few instances. The present multislice 1H-MRSI study shows that: (1) patients with schizophrenia, when compared as a group to normal controls, show a consistent 1H-MRSI pattern of group differences, i.e., bilateral reductions of NAA/CRE and NAA/CHO in HIPPO and DLPFC; (2)1H-MRSI data in both patients and controls do not show significant changes over this 90-day period; however, absolute metabolite ratios in individuals show low predictability over this time interval; (3) 1H-MRSI data show relatively low variability (as measured by the CVs) both in patients and normal controls, especially for NAA/CRE and CHO/CRE. PMID- 9408914 TI - Serotonin function and risk for alcoholism in boys with attention-deficit hyperactivity disorder. AB - Data in animals and adults indicate that central serotonergic (5-HT) function may be involved in the development of alcohol abuse. Despite this, studies exploring this mechanism in individuals at risk for alcoholism are scant. This study used a fenfluramine (FEN) challenge procedure to investigate the relationship between risk for alcoholism and 5-HT function in 7- to 11-year-old boys with attention deficit hyperactivity disorder (ADHD). The prolactin (PRL) and cortisol (CORT) responses to FEN were examined in 10 sons of alcoholic fathers (FA+) and 30 sons of nonalcoholic fathers (FA-). The FA+ group had a significantly greater CORT, but not PRL, response to FEN relative to the FA- group. The discrepancy between the CORT and PRL responses may be due to the different mechanisms that underlie their 5-HT stimulated release. This suggests that, among ADHD boys, those at familial risk for alcohol abuse may differ from those who are not at risk in 5-HT function. PMID- 9408915 TI - Glutamate modulation of dopamine measured in vivo with positron emission tomography (PET) and 11C-raclopride in normal human subjects. AB - Subanesthetic doses of the noncompetitive N-methyl-D-aspartate (NMDA) antagonist ketamine exacerbate psychosis in schizophrenic patients, and ketamine has significant abuse liability. These observations indicate that a secondary effect of ketamine may be to increase dopamine concentrations. The present study was undertaken using positron emission tomography (PET) and the dopamine (D2) radiotracer 11C-raclopride to determine whether ketamine would decrease D2 receptor availability, indicative of an increase in dopamine concentrations. Two scans were performed in seven male control subjects before and after administration of ketamine (0.5 mg/kg, i.v. infused over 20 min). Ketamine significantly increased cortisol levels and decreased dopamine receptor availability in the striatum (specific binding), but not in the cerebellum (nonspecific binding). In addition, the cerebellar binding subtracted from the striatal binding (to account for changes in nonspecific binding) was significantly decreased after ketamine administration. These results provide in vivo evidence for the ability of ketamine to increase striatal dopamine concentrations, consistent with the role of the NMDA receptor in modulating dopamine function. PMID- 9408916 TI - Dopamine receptor antagonists fail to prevent induction of cocaine sensitization. AB - We investigated the ability of dopamine D1 and D2 class receptor antagonists to prevent the induction of behavioral sensitization to cocaine. The D2 receptor antagonist eticlopride failed to prevent the induction of cocaine sensitization. An intermediate dose of the D1 receptor antagonist SCH 23390 (0.1 mg/kg) appeared to prevent the induction of cocaine sensitization when tested after 3 days of withdrawal, but sensitization was clearly evident after 10 days of withdrawal. High doses of SCH 23390 alone produced supersensitivity to the behavioral effects of cocaine and to the inhibitory effects of D1 receptor agonists on nucleus accumbens neurons. Co-administration of eticlopride and SCH 23390 also failed to prevent the induction of cocaine sensitization. SCH 23390, but not eticlopride, prevented the expression of cocaine sensitization. We conclude that dopamine receptors are either not involved in the induction of cocaine sensitization or that redundant mechanisms exist to produce the same neuroadaptations. PMID- 9408917 TI - Olanzapine versus placebo and haloperidol: quality of life and efficacy results of the North American double-blind trial. AB - This double-blind study evaluated the impact of treatment with olanzapine compared with haloperidol, and placebo on improvements in symptomatology and quality of life in patients with a DSM-III-R diagnosis of schizophrenia. A total of 335 patients was randomized to five treatment groups; olanzapine 5 +/- 2.5 mg/day, olanzapine 10 +/- 2.5 mg/day, olanzapine 15 +/- 2.5 mg/day, haloperidol 15 +/- 5 mg/day, and placebo. Patients responding to treatment during the 6-week acute phase were eligible to enter a 46-week extension. Efficacy measures included the brief psychiatric rating scale total, scale for assessment of negative symptoms summary, and clinical global impressions severity scores. Quality of life was evaluated using the quality of life scale. Data analyzed after 24 weeks of therapy showed that olanzapine was significantly superior to placebo in reducing clinical severity and significantly superior to haloperidol in reducing negative symptoms in patients responding to acute treatment. Furthermore, improvement in quality of life was observed in olanzapine-treated responders. PMID- 9408918 TI - Discrepant findings of clozapine effects on prepulse inhibition of startle: is it the route or the rat? AB - Studies examined methodological differences that might account for discrepant reports related to the ability of clozapine to restore prepulse inhibition (PPI) of acoustic startle in apomorphine (APO)-treated rats. Changes in PPI after APO and clozapine were compared in Sprague. Dawley (SD) versus Wistar rats. In SD rats, intraperitoneal administration of clozapine (4-12 mg/kg) completely reversed the PPI-disruptive effects of APO (0.5 mg/kg), with significant effects evident at the lowest dose of clozapine. Compared to SD rats, Wistars exhibited a relatively weaker (but statistically significant) disruption of PPI with the same or higher doses of APO and were also less sensitive to the PPI-restorative effects of clozapine. Clozapine administered via subcutaneous route completely restored PPI after APO treatment in SD rats. Discrepant findings with this model can be attributed to differences in rat strain; SD rats exhibit patterns of drug responses in this model that are optimal for examining profiles of putative atypical antipsychotics. PMID- 9408920 TI - Pattern recognition approaches in biomedical and clinical magnetic resonance spectroscopy: a review. AB - In recent years considerable effort has been devoted to applying pattern recognition techniques to the complex task of data analysis in magnetic resonance spectroscopy. It may be argued that such techniques will facilitate putting MRS technology to practical clinical use. This paper reviews approaches of pattern recognition commonly used in the analysis of MR spectra for biomedical applications. It briefly introduces the mathematical and algorithmic formulation of each of the techniques, noting their developmental background and their relationship to each other, and discusses their strengths and limitations. It then reviews how these techniques have been implemented in MRS applications. In doing so the paper also highlights a number of problems related to the design and testing of MRS/pattern recognition applications which currently prevent these techniques from being in wide practical clinical use, and suggests ways to avoid those pitfalls. PMID- 9408919 TI - Serotonergic agents that activate 5HT2A receptors prevent NMDA antagonist neurotoxicity. AB - Phencyclidine, ketamine, and other agents that block NMDA glutamate receptors trigger a schizophrenia-like psychosis in humans and induce pathomorphological changes in cerebrocortical neurons in rat brain. Accumulating evidence suggests that a complex network disturbance involving multiple transmitter receptor systems is responsible for the neuronal injury, and it is proposed that a similar network disturbance is responsible for the psychotomimetic effects of NMDA antagonists, and might also be involved in the pathophysiology of schizophrenia. In the present study we present evidence that serotonergic agents possessing 5HT2A agonist activity prevent NMDA antagonist neurotoxicity in rat brain. It is proposed that 5HT2A agonists may also prevent the psychotomimetic effects of NMDA antagonists. Among the 5HT2A agonists examined and found to be neuroprotective are LSD and related hallucinogens. The apparent contradiction in proposing that these agents might have antipsychotic properties is resolved by evidence linking their hallucinogenic activity to agonist action at 5HT2C receptors, whereas antipsychotic activity would be attributable to agonist action at 5HT2A receptors. PMID- 9408921 TI - Retrospective correction of surface coil MR images using an automatic segmentation and modeling approach. AB - The use of surface coils in magnetic resonance imaging offers significant improvements in the signal-to-noise ratio over volume coils for many applications. However, the inhomogeneous reception profile of surface coils hampers their usefulness by introducing significant nonuniformities or intensity variations which can vary by greater than six-fold across the sample. In this study, we evaluated an automatic technique for retrospective correction of intensity variations observed in a high-resolution surface coil MR image of the rat brain obtained using an adiabatic magnetic resonance imaging sequence. The images are shown to have a coefficient of variation less than 12% following application of this correction algorithm. This image intensity correction technique can be applied retrospectively to all data sets and corrects both sample/patient dependent effects (e.g. attenuation of overlying tissue) or sample independent effects (e.g. coil geometry or position). This approach should also prove valuable in improving regions of interest analysis, volume histograms and thresholding techniques. PMID- 9408922 TI - Free magnesium-ion concentration in erythrocytes by 31P NMR: the effect of metabolite-haemoglobin interactions. AB - The effects that haemoglobin-metabolite interactions have on estimates of free magnesium-ion concentration in human erythrocytes, determined by 31P NMR [Gupta, R. K. et al., J. Biol. Chem. 253, 6172-6176 (1978)], were investigated. If the metabolite-haemoglobin association constants of Berger et al. [Eur. J. Biochem. 38, 553-562 (1973)] are used in the analysis then the estimates of intracellular free magnesium-ion concentration made by Gupta et al. (0.25 and 0.67 mM) become 0.43 and 0.60 mM, for oxygenated and deoxygenated cells, respectively. In oxygenated cells, this difference is primarily due to the lower value of KHbMgATP, given by Berger et al. These newly calculated concentrations are in closer agreement with those of Flatman (0.40 mM for oxygenated cells; 0.62 mM for deoxygenated cells) [Flatman, P. W., J. Physiol. 300, 19-30 (1980)] obtained with the 'zero-point titration' method. In addition, the assumptions that the chemical shift separations between the alpha- and beta-phosphorus resonances of ATP and MgATP are unchanged on association with Hb were shown to be false. Under normal intracellular conditions this may lead to errors of 5-10%. Much larger errors would be possible in cases where significant amounts of ATP or MgATP are bound to Hb. These outcomes place doubt on measurements of intracellular free Mg2+ concentration made using 31P NMR if there is no consideration given to the total concentration of 2,3-bisphosphoglycerate (BPG), ATP and Hb in the sample; the same principle would apply to other cell-types. PMID- 9408923 TI - PET and NMR dual acquisition (PANDA): applications to isolated, perfused rat hearts. AB - Positron emission tomography and nuclear magnetic resonance spectroscopy are non invasive techniques that allow serial metabolic measurements to be obtained in a single subject. Significant advantages could be obtained if both types of scans could be acquired with a single machine. A small-scale PET scanner, designed to operate in a high magnetic field, was therefore constructed and inserted into the top half of a 7.3 cm bore, 9.4 T NMR magnet and its performance characterized. The magnetic field did not significantly affect either the sensitivity (approximately 3 kcps/MBq) or the spatial resolution (2.0 mm full width at half maximum, measured using a 0.25 mm diameter line source) of the scanner. However, the presence of the PET scanner resulted in a small decrease in field homogeneity. The first, simultaneous 31P NMR spectra (200, 80 degrees pulses collected at 6 s intervals) and PET images (transverse, mid-ventricular slices at the level of the mitral value) from isolated, perfused rat hearts were acquired using a specially designed NMR probe inserted into the bottom half of the magnet. The PET images were of excellent quality, enabling the left ventricular wall and interventricular septum to be clearly seen. In conclusion, we have demonstrated the simultaneous acquisition of PET and NMR data from perfused rat hearts; we believe that the combination of these two powerful techniques has tremendous potential in both the laboratory and the clinic. PMID- 9408925 TI - Current awareness in NMR in biomedicine. PMID- 9408924 TI - On the correlation between tissue hydration state and proton NMR relaxation rates in experimental liver transplantation. AB - As the tissue hydration state is one of the most important parameters to predict viability cold stored livers before transplantation, we investigated the correlation between the tissue inverse total water fraction, reflecting the hydration state, and proton relaxation times in cold stored rat liver and orthotopic liver transplantation in a pig model. In cold stored rat liver excellent linear correlations between relaxation rates R1 (= 1/T1) and R2 (= 1/T2) and inverse total water fraction 1/Pw were obtained. In pig liver transplants, the slope and intercept obtained from a linear regression model are twice as high for R1 and almost identical for R2; however, correlation coefficients are lower due to increased biological variation and a smaller range in storage conditions, reflected by the range of water content. Proton nuclear magnetic resonance relaxation times measured during the cold storage on the whole organ non-invasively show also linear correlation with the inverse total water fraction, but the method is presently not accurate enough to estimate the hydration state of the liver tissue with sufficient precision. NMR relaxation times obtained from liver biopsies have the potential to predict tissue viability in experimental liver transplantation independent of species, strain and gender, and thus may be useful in estimating the viability of human donor livers (or at least add a new complementary information to the information gained by standard liver selection and function test before and after transplantation). PMID- 9408926 TI - Listeriosis and pregnancy: food for thought. PMID- 9408927 TI - Pregnancy-induced hypertension: genesis of and response to endothelial injury and the role of endothelin 1. AB - This paper discusses the spectrum of pregnancy-induced hypertension and presents a theory for its etiology. Endothelial injury is the purported precursor to pregnancy-induced hypertensive disorders, and this discussion expands on a possible mechanism by which injury could occur as a result of incomplete trophoblastic invasion. We review endothelin physiology and compare and contrast the evidence surrounding endothelin 1 as a putative mediator of PIH. An approach to treatment utilizing antagonists to the endothelin 1 receptor is introduced. PMID- 9408928 TI - Delivery of the nonvertex second twin: a review of the literature. AB - Twin gestations comprise approximately 1 percent of all pregnancies (1), and are associated with increased perinatal morbidity and mortality, mainly due to the increased incidence of prematurity and growth restriction (2). Hazards of twin delivery can be attributed to malpresentation, most often by the second twin. The vertex-nonvertex presentation occurs in approximately 40 percent of all twins (3, 4). Although there is consensus regarding the safety of vaginal delivery for twins when both are vertex (5), controversy exists over intrapartum management when the second twin is nonvertex. Some investigators advocate cesarean delivery, particularly when the second twin is nonvertex (6), or if the expected birth weight is < 2000 gm (6-8). This review aims to determine whether vaginal delivery of the nonvertex second twin, either by breech extraction or attempted external cephalic version, is associated with increased morbidity or mortality over cesarean delivery. PMID- 9408929 TI - Arteriovenous malformations of the uterus: an uncommon cause of vaginal bleeding. AB - Arteriovenous malformations (AVM) are rare entities in gynecology, with only 73 cases reported in the literature. Most commonly they present with vaginal hemorrhage, but other presentations such as congestive heart failure, postmenopausal bleeding, and an asymptomatic mass have been described. These lesions may be congenital or acquired. Acquired lesions are believed to follow trauma or may arise after choriocarcinoma or other gynecologic malignancies. Diagnosis can rapidly be made with color flow Doppler ultrasound or angiography. Additionally, they have been detected using hysteroscopy, hysterosalpingogram, and computerized tomography. Acute management consists of hemodynamic stabilization and possibly placement of a Foley bulb in the uterus or methylergonovine injection. Ultimate treatment depends on the patients desire for fertility. Embolization therapy is variably successful and may allow the preservation of reproductive capacity. To date, five pregnancies after embolization have been reported with varying outcomes. If pregnancy is not desired or embolization fails, hysterectomy remains the treatment of choice. PMID- 9408930 TI - The patient as decision maker. PMID- 9408931 TI - Keep your hands off? PMID- 9408932 TI - Group hydrotherapy versus group land-based treatment for chronic low back pain. AB - Sixty subjects with chronic low back pain (LBP) were sequentially allocated to either hydrotherapy treatment or land treatment groups in order of presentation. Subjects acted as their own controls for a period of three weeks, after which they attended their respective group sessions twice weekly for six weeks. Twenty eight subjects from each group attended all treatment and assessment sessions. Results indicated that both groups improved significantly in functional ability and in decreasing pain levels. Thoracolumbar mobility did not improve significantly in either group. Overall there was no significant difference found between the two types of treatment, although results should be viewed as encouraging for the advocates of both hydrotherapy and land-based exercise as a treatment for chronic LBP. PMID- 9408933 TI - Does ice immersion influence ankle joint position sense? AB - The purpose of this study was to determine whether a fifteen minute ice immersion treatment influenced the normal ankle joint position sense at 40% and 80% range of inversion and to establish the length of treatment effect through monitoring the rewarming process. Forty nine healthy volunteers between the ages of 17 and 28 were tested. Subjects were screened to exclude those with a history of ankle injuries. The subject's skin temperature over antero-lateral aspect of the ankle was measured using a thermocouple device during the fifteen minutes ice intervention and thirty minutes post-intervention. Testing of ankle joint position sense using the pedal goniometer was performed before and after a clinical application of ice immersion. The testing required the subject to actively reposition their ankle at 40% and 80% of their total range of inversion. The majority of subjects experienced numbness of the foot and ankle by the fifth or sixth minute during ice immersion. One minute after immersion skin temperatures averaged 15 degrees C + 1.7 degrees C. Skin temperature was seen to rise relatively rapidly for the first ten minutes and then slowed considerably. Subjects had not returned to the pre-test skin temperatures by thirty minutes. A significant difference in ankle joint position sense (p < 0.0499) following fifteen minutes of ice immersion was found. However, the magnitude of this difference (0.5 degree) would not be deemed significant in clinical practice. The research found no significant difference in joint position sense between 40% and 80% of the range of inversion both before and after cryotherapy. These findings suggest that the clinical application of cryotherapy is not deleterious to joint position sense and assuming normal joint integrity patients may resume exercise without increased risk of injury. PMID- 9408934 TI - A multidimensional scaling analysis of the techniques that physiotherapists use. AB - Following the essentially 'hands-on' origin of the physiotherapy profession a century ago, the range of techniques available to practitioners has expanded considerably. While this expansion of techniques has been of research interest, the main focus has been upon individual techniques, thus neglecting the fundamental question of the use of techniques in combination. In addition, researchers have tended to treat the profession as homogeneous, thus overlooking possible differences in technique usage. The present study addressed these questions. It postulated that ordered combinations of techniques usage would be employed, and that a typology would exist that differentiates the profession according to patterns of use. The study comprised a questionnaire survey of 230 clinical physiotherapists in England. This sought information on the range and frequency of techniques used over the previous six months. Descriptive analysis indicated a high frequency of use of several techniques, notably exercise therapy, passive mobilisation techniques and ultrasound. A feature of the analysis was the use of two multivariate procedures, multidimensional scalogram analysis (MSA) and smallest space analysis (SSA). The SSAs revealed a clear structure concerning combinations of techniques used, and the MSAs provided a coherent typology based upon this usage. One feature of the techniques in combination that emerged was the frequent use of passive mobilising techniques, which was associated with equally frequent use of electrotherapy techniques. In contrast, respondents who employed neurological or respiratory techniques made little use of electrotherapy. Regarding the typology, differentiation of the profession according to use of particular combinations of techniques indicated probable specialisms. The specialist areas identified were neurological physiotherapy, respiratory physiotherapy and musculo-skeletal physiotherapy. The results reinforce the need for both future research and physiotherapy education to address the issue of techniques in combination. Finally, and on the basis of the analyses, elements of a theoretical model are advanced that may provide directions for future research. PMID- 9408935 TI - Stimulation of bone healing in new fractures of the tibial shaft using interferential currents. AB - The aim of this research was to establish whether interferential currents (IFC) could significantly reduce the healing time for fractures of the tibia and thereby prevent nonunion. Males between the ages of 12 and 86, who had sustained fractures of the tibiae, were entered into this double blind clinical trial. According to strict inclusion and exclusion criteria, a final sample of 227 cases (208 subjects) were entered by block randomization into two groups; an experimental group (n = 41) and placebo group (n = 35). A further group was entered retrospectively--control group (n = 151). IFCs were applied to the experimental group via suction electrodes for 30 minutes per day for 10 days, using a beat frequency of 10-25 Hz and a swing mode of 6 integral of 6. The placebo group had only suction electrodes applied, the intermittent mode produced a rhythmical massage effect; subjects in this group commented on pain relief which resulted in the addition of the control group as a check on the possible effect of intermittent suction. The control group received no intervention. The data were analysed using analysis of covariance which resulted in a finding of no significant difference in the time taken to union for the three groups. This means that to date there is no reason to believe that IFCs (using the parameters of this trial) can reduce the healing time for new fractures of the tibia or prevent nonunion. However, further investigation is recommended. PMID- 9408936 TI - Evidence based medicine. PMID- 9408937 TI - Bases of practice in neurological physiotherapy. PMID- 9408938 TI - Mechanisms of signaling and related enzymes. AB - Most enzymes involved in cell signaling, such as protein kinases, protein phosphatases, GTPases, and nucleotide cyclases catalyze nucleophilic substitutions at phosphorus. When possible, the mechanisms of such enzymes are most clearly described quantitatively in terms of how associative or dissociative they are. The mechanisms of cell signaling enzymes range from < or = 8% associative (cAMP-dependent protein kinase) to approximately 50% associative (G protein Gi alpha 1). Their catalytic powers range from 10(5.7) (p21ras) to 10(11.7) (lambda Ser-Thr protein phosphatase), usually comparable in magnitude with those of nonsignaling enzymes of the same mechanistic class. Exceptions are G proteins, which are 10(3)- to 10(5)-fold poorer catalysts than F1 and myosin ATPases. The lower catalytic powers of G proteins may be ascribed to the absence of general base catalysis, and additionally in the case of p21ras, to the absence of a catalytic Arg residue, which interacts with the transition state. From kinetic studies of mutant and metal ion substituted enzymes, the catalytic powers of cell signaling and related enzymes can be rationalized quantitatively by factors contributed by metal ion catalysis (> or = 10(5), general acid catalysis (approximately 10(3 +/- 1)), general base catalysis (approximately 10(3 +/- 1)), and transition-state stabilization by cationic and hydrogen bond donating residues (approximately 10(3 +/- 1)). PMID- 9408939 TI - Chaos in protein dynamics. AB - MD simulations, currently the most detailed description of the dynamic evolution of proteins, are based on the repeated solution of a set of differential equations implementing Newton's second law. Many such systems are known to exhibit chaotic behavior, i.e., very small changes in initial conditions are amplified exponentially and lead to vastly different, inherently unpredictable behavior. We have investigated the response of a protein fragment in an explicit solvent environment to very small perturbations of the atomic positions (10(-3) 10(-9) A). Independent of the starting conformation (native-like, compact, extended), perturbed dynamics trajectories deviated rapidly, leading to conformations that differ by approximately 1 A RMSD within 1-2 ps. Furthermore, introducing the perturbation more than 1-2 ps before a significant conformational transition leads to a loss of the transition in the perturbed trajectories. We present evidence that the observed chaotic behavior reflects physical properties of the system rather than numerical instabilities of the calculation and discuss the implications for models of protein folding and the use of MD as a tool to analyze protein folding pathways. PMID- 9408940 TI - Highly constrained multiple-copy refinement of protein crystal structures. AB - In the course of refining atomic protein structures, one often encounters difficulty with molecules that are unusually flexible or otherwise disordered. We approach the problem by combining two relatively recent developments: simultaneous refinement of multiple protein conformations and highly constrained refinement. A constrained Langevin dynamics refinement is tested on two proteins: neurotrophin-3 and glutamine synthetase. The method produces closer agreement between the calculated and observed scattering amplitudes than standard, single copy, Gaussian atomic displacement parameter refinement. This is accomplished without significantly increasing the number of fitting parameters in the model. These results suggest that loop motion in proteins within a crystal lattice can be extensive and that it is poorly modeled by isotropic Gaussian distributions for each atom. PMID- 9408941 TI - Small-angle X-ray scattering and crystallographic studies of arcelin-1: an insecticidal lectin-like glycoprotein from Phaseolus vulgaris L. AB - Arcelin-1 and alpha-amylase inhibitor are two lectin-like glycoproteins expressed in the seeds of the kidney bean (Phaseolus vulgaris). They display insecticidal activities and protect the seeds from predation by larvae of various bruchids through different biological actions. Solution-state investigations by small angle X-ray scattering (SAXS) show the dimeric structure of arcelin-1, a requirement for its hemagglutinating properties. Anions were found to have specific properties in their effectiveness to disrupt protein aggregates, affect solubility, and improve crystallizability. The SAXS results were used to improve crystallization conditions, and single crystals diffracting beyond 1.9 A resolution were obtained. X-ray diffraction data analysis shows that noncrystallographic symmetry-related arcelin-1 molecules form a lectin-like dimer and reveals the presence of a solvent-exposed anion binding site on the protein, at a crystal-packing interface. The solution state properties of arcelin-1 and crystal twinning may be explained by the anion specificity of this binding site. PMID- 9408942 TI - Improved method for prediction of protein backbone U-turn positions and major secondary structural elements between U-turns. AB - A new and more accurate method has been developed for predicting the backbone U turn positions (where the chain reverses global direction) and the dominant secondary structure elements between U-turns in globular proteins. The current approach uses sequence-specific secondary structure propensities and multiple sequence information. The latter plays an important role in the enhanced success of this approach. Application to two sets (total 108) of small to medium-sized, single-domain proteins indicates that approximately 94% of the U-turn locations are correctly predicted within three residues, as are 88% of dominant secondary structure elements. These results are significantly better than our previous method (Kolinski et al., Proteins 27:290-308, 1997). The current study strongly suggests that the U-turn locations are primarily determined by local interactions. Furthermore, both global length constraints and local interactions contribute significantly to the determination of the secondary structure types between U-turns. Accurate U-turn predictions are crucial for accurate secondary structure predictions in the current method. Protein structure modeling, tertiary structure predictions, and possibly, fold recognition should benefit from the predicted structural data provided by this new method. PMID- 9408943 TI - Evolution of model proteins on a foldability landscape. AB - We model the evolution of simple lattice proteins as a random walk in a fitness landscape, where the fitness represents the ability of the protein to fold. At higher selective pressure, the evolutionary trajectories are confined to neutral networks where the native structure is conserved and the dynamics are non self averaging and nonexponential. The optimizability of the corresponding native structure has a strong effect on the size of these neutral networks and thus on the nature of the evolutionary process. PMID- 9408944 TI - New insights into the molecular basis of lectin-carbohydrate interactions: a calorimetric and structural study of the association of hevein to oligomers of N acetylglucosamine. AB - Isothermal titration calorimetry was used to characterize thermodynamically the association of hevein, a lectin from the rubber tree latex, with the dimer and trimer of N-acetylglucosamine (GlcNAc). Considering the changes in polar and apolar accessible surface areas due to complex formation, we found that the experimental binding heat capacities can be reproduced adequately by means of parameters used in protein-unfolding studies. The same conclusion applies to the association of the lectin concanavalin A with methyl-alpha-mannopyranoside. When reduced by the polar area change, binding enthalpy values show a minimal dispersion around 100 degrees C. These findings resemble the convergence observed in protein-folding events; however, the average of reduced enthalpies for lectin carbohydrate associations is largely higher than that for the folding of proteins. Analysis of hydrogen bonds present at lectin-carbohydrate interfaces revealed geometries closer to ideal values than those observed in protein structures. Thus, the formation of more energetic hydrogen bonds might well explain the high association enthalpies of lectin-carbohydrate systems. We also have calculated the energy associated with the desolvation of the contact zones in the binding molecules and from it the binding enthalpy in vacuum. This latter resulted 20% larger than the interaction energy derived from the use of potential energy functions. PMID- 9408945 TI - SHBG region of the anticoagulant cofactor protein S: secondary structure prediction, circular dichroism spectroscopy, and analysis of naturally occurring mutations. AB - Protein S (PS) and growth arrest specific factor 6 (GAS6) are vitamin K-dependent proteins with similar structures. They are mosaic proteins possessing a carboxyl terminal region presenting sequence similarity with plasma sex hormone binding globulin (plasma SHBG), although apparently not involved in steroid binding. The SHBG-like modules have sequence similarity with the G repeats of the chain A of laminin. Laminin G repeats have been reported to contain mainly beta-strands (about 40-50%) but no or little alpha structure by circular dichroism (CD) spectroscopy. Secondary structure predictions carried out in the present work unexpectedly showed a 20 to 27% helices content in the SHBG region of PS/GAS6 (about 100 residues), while plasma SHBG and laminin G repeats had around 10% helices. CD measurements for human PS indicated also that its SHBG region had about 100 residues in alpha-helical structure. These data suggest that the SHBG region of PS/GAS6 on the one hand, and the laminin G repeats and possibly plasma SHBG on the other hand, could present important structural differences. Previously reported polymorphisms and point mutations leading to PS deficiency and thrombophilia have been analyzed with our structural predictions. We found a good agreement between these structural predictions, CD measurements, experimental and clinical data. This information allows us to gain insights into the three-dimensional structure of PS that will be helpful for the design of new experiments and future clinical investigations. PMID- 9408946 TI - Effects of organic solvents on protein structures: observation of a structured helical core in hen egg-white lysozyme in aqueous dimethylsulfoxide. AB - A partly folded state of hen egg-white lysozyme has been characterized in 50% DMSO. Low concentrations of DMSO (< 10%) have little effect on the overall folded conformation of lysozyme as seen from 1H NMR chemical shift dispersion. At increasing DMSO concentrations (> 10%) a cooperative transition of the structure to a new, partially folded state is observed. This transition is essentially complete by approximately 50% DMSO. NMR studies show an overall decrease in chemical shift dispersion with marked broadening of many resonances. A substantial number of backbone and side chain-side chain NOEs suggests the presence of secondary and tertiary interactions in the intermediate state. Tertiary organization of the aromatic residues is also demonstrated by enhanced near-UV circular dichroism and limited exposure of tryptophans as monitored by iodide quenching of fluorescence. The intermediate state exhibits enhanced binding to hydrophobic dyes. Further, the structural transition from this state to a largely unfolded conformation is cooperative. H/D exchange rates of several amide protons and four indole protons of tryptophans (W28, W108, W111, and W123), measured by refolding from 50% DMSO at different time intervals reveal that protection factors are high for the helical domain, whereas NH groups in the triple stranded antiparallel beta-sheet domain are largely solvent-exposed. An ordered hydrophobic core in the intermediate state comprising of helix A, helix B, and helix D is consistent with the high protection factors observed. The structured intermediate in 50% DMSO resembles the early kinetic intermediate observed in the refolding of hen egg white lysozyme, as well as a molten globule state of equine lysozyme at low pH. The results demonstrate the potential use of non-aqueous structure perturbing solvents like DMSO to stabilize partially folded conformations of proteins. PMID- 9408947 TI - Monte Carlo simulation of protein folding with orientation-dependent monomer monomer interactions. AB - We present the results of lattice Monte Carlo simulations of protein folding in the framework of a model taking into account (i) the dependence of the energy of interaction of amino-acid residues on their orientation and (ii) the rigidity of the polypeptide chain with respect to the formation of kinks. If the chain is flexible, the final protein structures are predicted to be compact. Increasing the energy cost of creation of kinks is found to favor the formation of flat structures mimicking an ideal antiparallel beta sheet. For compact structures, the kinetics of folding exhibit the standard two-phase regime (a rapid collapse to one of the metastable stable, followed by slow reconfiguration of the chain to the native structure). For flat structures, the transition to the native state is often gradual. PMID- 9408948 TI - Species-specificity of the cohesin-dockerin interaction between Clostridium thermocellum and Clostridium cellulolyticum: prediction of specificity determinants of the dockerin domain. AB - The cross-species specificity of the cohesin-dockerin interaction, which defines the incorporation of the enzymatic subunits into the cellulosome complex, has been investigated. Cohesin-containing segments from the cellulosomes of two different species, Clostridium thermocellum and Clostridium cellulolyticum, were allowed to interact with cellulosomal (dockerin-containing) enzymes from each species. In both cases, the cohesin domain of one bacterium interacted with enzymes from its own cellulosome in a calcium-dependent manner, but the same cohesin failed to recognize enzymes from the other species. Thus, in the case of these two bacteria, the cohesin-dockerin interaction seems to be species specific. Based on intra- and cross-species sequence comparisons among the different dockerins together with their known specificities, we tender a prediction as to the amino-acid residues critical to recognition of the cohesins. The suspected residues were narrowed down to only four, which comprise a repeated pair located within the calcium-binding motif of two duplicated sequences, characteristic of the dockerin domain. According to the proposed model, these four residues do not participate in the binding of calcium per se; instead, they appear to serve as recognition codes in promoting interaction with the cohesin surface. PMID- 9408949 TI - Theoretical investigation of IL-6 multiprotein receptor assembly. AB - Interleukin-6 (IL-6) is a multifunctional cytokine that regulates cell growth, differentiation, and cellular functions in many cell lineages. Recently, evidences for the formation of an active hexameric complex with an IL-6:IL-6R alpha:gp130 stoichiometry of 2:2:2 have been obtained by different experimental approaches. Analysis of the electrostatic potential complementarity between IL-6 and its receptors has been used, in this study, to guide the assembly of homology based 3D models of the components. The results strongly support a mechanism whereby the active cytokine (IL-6: IL-6R alpha) associates with the signal transducing gp130 protein, and the trimeric complex formed further dimerizes to form the hexameric species. Furthermore, computational simulations of the multiprotein complexes provide a rationalization of data from mutation experiments and highlight some key protein-protein interactions which have not yet been the subject of mutagenesis studies. PMID- 9408950 TI - SH3 domain of Bruton's tyrosine kinase can bind to proline-rich peptides of TH domain of the kinase and p120cbl. AB - X-linked agammaglobulinemia (XLA), an inherited disease, is caused by mutations in the Bruton's tyrosine kinase (BTK). The absence of functional BTK leads to failure of B-cell differentiation; this incapacitates antibody production in XLA patients, who suffer from recurrent, sometimes lethal, bacterial infections. BTK plays an important role in B-cell development; it interacts with several proteins in the context of signal transduction. Point mutation in the BTK gene that leads to deletion of C-terminal 14 aa residues of BTK SH3 domain was found in a patient family. To understand the role of BTK, we studied binding of BTK SH3 domain (aa 216-273, 58 residues) and truncated SH3 domain (216-259, 44 residues) with proline-rich peptides; the first peptide constitutes the SH3 domain of BTK, while the latter peptide lacks 14 amino acid residues of the C terminal. Proline-rich peptides selected from TH domain of BTK and p120cbl were studied. It is known that BTK TH domain binds to SH3 domains of various proteins. We found that BTK SH3 domain binds to peptides of BTK TH domain. This suggests that BTK SH3 and TH domains may associate in inter- or intramolecular fashion, which raises the possibility that the kinase may be regulating its own activity by restricting the availability of both its ligand-binding modules. We also found that truncated SH3 domain binds to BTK TH domain peptide less avidly than does normal SH3 domain. Also, we show that the SH3 and truncated SH3 domains bind to peptide of p120cbl, but the latter domain binds weakly. It is likely that the truncated SH3 domain fails to present to the ligand the crucial residues in the correct context, hence the weaker binding. These results delineate the importance of C-terminal in binding of SH3 domains and indicate also that improper folding and the altered binding behavior of mutant BTK SH3 domain likely leads to XLA. PMID- 9408951 TI - A combinatorial library for the binuclear metal center of bacterial phosphotriesterase. AB - Phosphotriesterase (PTE) is a zinc metalloenzyme that catalyzes the hydrolysis of an extensive array of organophosphate pesticides and mammalian acetylcholinesterase nerve agents. Although the three-dimensional crystal structure of PTE has been solved (M. M. Benning et al., Biochemistry 34:7973 7978, 1995), the precise functions of the individual amino acid residues that interact directly with the substrate at the active site are largely unknown. To construct mutants of PTE with altered specificities for particular target substrates, a simple methodology for generating a library of mutants at specific sites was developed. In this investigation, four of the six protein ligands to the binuclear metal site (His-55, His-57, His-201, and His-230) were targeted for further characterization and investigation. Using the polymerase chain reaction (PCR) protocols, a library of modified PTE genes was generated by simultaneously creating random combinations of histidine and cysteine codons at these four positions. The 16 possible DNA sequences were isolated and confirmed by dideoxy DNA sequencing. The 16 mutant proteins were expressed in Escherichia coli and grown with the presence or absence of 1 mM CoCl2, ZnSO4, or CdSO4 in the growth medium. When grown in the presence of CoCl2, the H57C protein cell lysate showed greater activity for the hydrolysis of paraoxon than the wild type PTE cell lysate. H201C and H230C exhibited up to 15% of the wild-type activity, while H55C, a green protein, was inactive under all assay conditions. All other mutants had < 10(-5) of wild-type activity. None of the purified mutants that exhibited catalytic activity had a significantly altered Km for paraoxon. PMID- 9408952 TI - Primary structure of the common polypeptide chain b from the multi-hemoglobin system of the hydrothermal vent tube worm Riftia pachyptila: an insight on the sulfide binding-site. AB - The deep-sea tube worm Riftia pachyptila Jones possesses a multi-hemoglobin system with three different extracellular Hbs: two dissolved in the vascular blood, V1 (ca. 3,500 kDa) and V2 (ca. 400 kDa), and one in the coelomic fluid, C1 (ca. 400 kDa). V1 Hb consists of four heme-containing, globin chains (b-e) and four linker chains (L1-L4). V2 and C1 Hbs are exclusively built from globin chains, six for V2 (a-f) and five for C1 (a-e). The complete amino acid sequence of the isolated monomeric globin chain b, common to all Riftia Hbs, has been determined by automated Edman degradation sequencing of the peptides derived by digestion with trypsin, chymotrypsin, thermolysin, and CNBr. This polypeptide chain is composed of 144 amino acid residues, providing a M(r) of 16, 135.0 Da. Moreover, the primary sequence of chain b revealed 3 Cys residues at position 4, 75, and 134. Cys-4 and Cys-134 are located at positions where an intra-chain disulfide bridge is formed in all annelid, vestimentiferan, or pogonophoran chains, but Cys-75 is located at a unique position only found in three globin chains belonging to Lamellibrachia and Oligobrachia, a vestimentiferan and a pogonophoran. In both groups, Hbs can bind sulfide reversibly to fuel the chemosynthetic process of the symbiotic bacteria they harbor. Sulfide-binding experiments performed on purified Hb fractions (i.e., V1, V2, and C1 Hbs) suggest that free Cys residues on globin chains, and the numerous Cys found in linker chains, as determined previously by ESI-MS, may be the sulfide binding-sites. Blocking the free Cys by N-ethylmaleimide, we confirmed that free cysteines were involved in sulfide-binding but did not account for the whole sulfide-binding capacity of V1 Hb. Furthermore, a phylogenetic tree was constructed from 18 globin-like chains of annelid, vestimentiferan, and pogonophoran extracellular Hbs to clarify the systematic position of tubeworms. Riftia chain b clearly belongs to the "strain A" family with 30 to 80% identity with the other sequences analyzed. Its position in the tree confirmed a close relationship between vestimentiferan, pogonophoran, and annelid Hbs. PMID- 9408953 TI - Chain conformation in polyretropeptides III: design of a 3(10) helix using alpha,alpha-dialkylated amino acids and retropeptide bonds. AB - Computer simulations have been used to design a polypeptide with a 3(10) helix conformation. The study has been been performed taking advantage of the intrinsic helix forming tendency of alpha-Aminoisobutyric acid. In order to avoid the formation of the alpha helix, which is the other common helical conformation adopted by alpha-Aminoisobutyric acid-based peptides, retropeptide bonds have been included in the sequence. Thus, retropeptides are not able to form the intramolecular hydrogen bonding interactions characteristic of the alpha helix. The influences of both the peptide length and the solvent have been examined and compared with those of the polypeptide without retropeptide bonds. PMID- 9408954 TI - Orthopedic and interventional applications at low field MRI with horizontally open configuration. A review. AB - The recently introduced horizontally open configuration imagers allow imaging of knee, hip or shoulder during whole range of motion, which is not possible in conventional MR imagers. Special joint motion devices can be used to provide accurate and reproducible studies. In cervical spine, functional MR imaging may be useful in evaluating alarligament stability in patients with late sequelae of a whiplash injury, and in patients with rheumatoid arthritis who are clinically suspected of having a cervical myelopathy or superior migration of the odontoid process. In shoulder, full range of motion abduction study may be helpful in assessing the supraspinatus tendon impingement. To evaluate patellofemoral malalignment, quadriceps loading is recommended since associated contracting muscles and related soft tissue structures can be evaluated. The position of the femoral head relative to the acetabulum during different positions can be assessed. Open-configuration scanners provide an access to patients during scanning procedure, and therefore permit interventional procedures to be monitored with MRI. Such interventions include aspiration cytology/biopsy and different drainage procedures. PMID- 9408955 TI - Mechanisms of drug resistance in hematologic malignancies. AB - Multiple cellular mechanisms contribute to the overall clinical drug-resistant phenotype of malignant cells. A major mechanism of drug resistance documented to occur in hematologic malignancies is overexprssion of the MDR-1 gene product, P glycoprotein (P-gp). Drugs, called chemosensitizers, have been designed to overcome P-gp-mediated drug resistance, and these agents are now being tested in the clinic. Overcoming P-gp-mediated resistance may select for alternative mechanisms of resistance that are not affected by chemosensitizing agents. Alternative mechanisms are now being described, and the clinical relevance of these mechanisms is being investigated in hematologic malignancies. The exact mechanisms involved in the overall drug-resistant phenotype will likely depend on the type of malignancy and its exposure to anticancer drugs. A major challenge in improving the treatment of patients with hematologic malignancies will be to determine if and when these various cellular mechanisms contribute to clinical drug resistance. PMID- 9408956 TI - Bcl-2 family proteins: regulators of apoptosis and chemoresistance in hematologic malignancies. AB - Nearly all anticancer drugs presently available to clinicians kill tumor cells by activating an endogenous biochemical pathway for cell suicide, known as programmed cell death or apoptosis. However, many malignant cells develop defects in the regulation of genes that control apoptosis, rendering them resistant to the induction of apoptosis by a wide variety of stimuli, including chemotherapeutic drugs and radiotherapy. The BCL-2 family of proto-oncogenes are critical regulators of apoptosis whose expression frequently becomes altered in human cancers, including some of the most common types of lymphomas and leukemias. The first member of this gene family to be identified was BCL-2, by virtue of its involvement in t(14;18) chromosomal translocations commonly found in B-cell non-Hodgkin's lymphoma (NHL). Subsequently, however, overexpression of Bcl-2 has been reported in a wide variety of cancers, without associated chromosomal translocations. A variety of experiments have provided conclusive evidence that elevations in Bcl-2 expression cause resistance to chemotherapeutic drugs, while decreases in Bcl-2 expression promote apoptotic responses to anticancer drugs. Information about the biochemical mechanisms of action of the Bcl-2 protein and other members of the Bcl-2 family is beginning to suggest strategies for overcoming the cytoprotective effects of Bcl-2 overexpression in lymphomas, leukemias, and other types of cancer. PMID- 9408957 TI - Non-P-glycoprotein drug export mechanisms of multidrug resistance. AB - A variety of cellular mechanisms of multidrug resistance (MDR) have been identified in human drug-resistant cell lines, and may play an important role in the clinical response of hematologic malignancies to chemotherapy. P-glycoprotein (P-gp)-mediated drug efflux is the most well-characterized cellular mechanism of MDR; however, several other non-P-gp membrane transporter proteins have also been implicated in the development of an MDR phenotype in hematologic malignancies. These include the MDR-related protein (MRP), the lung-resistance protein (LRP), and the transporter of antigenic peptides (TAP). The transporter proteins MRP and TAP are both members of the adenosine triphosphate (ATP)-binding cassette (ABC) family of transmembrane transporters, but each has distinct differences in substrate specificity. Despite effective modulation of P-gp, one or more of these alternate mechanisms of drug resistance may contribute to an MDR phenotype in tumor cell lines. Development of multifunctional MDR modulators or novel therapeutics may be necessary to effectively circumvent MDR in hematologic malignancies. PMID- 9408958 TI - The prognostic significance of the expression and function of multidrug resistance transporter proteins in acute myeloid leukemia: studies of the Southwest Oncology Group Leukemia Research Program. AB - Resistance to chemotherapy is a major obstacle in the treatment of patients with acute myeloid leukemia (AML). The majority of AML patients are intrinsically resistant to chemotherapy at initial diagnosis before chemotherapeutic exposure; such intrinsic resistance frequently results from expression of the multidrug resistance gene (MDR-1), which encodes a membrane transporter protein, P glycoprotein (P-gp), that mediates drug efflux in leukemic cells. Expression of novel transporter proteins that confer alternative forms of multidrug resistance (MDR), such as the lung-resistance protein (LRP) and the MDR-associated protein (MRP), occurs more frequently in leukemic patients at relapse. Preliminary studies indicate that these proteins may also confer therapeutic resistance in leukemia. In patients older than 55 years of age, AML is characterized by a high frequency of unfavorable cytogenetics, P-gp expression, and functional drug efflux, which contribute to poor clinical outcome. In a multivariate analysis, secondary AML, unfavorable cytogenetics, and P-gp expression/function were each significantly and independently associated with a lower complete remission (CR) rate. Resistant disease was associated with P-gp expression and unfavorable cytogenetics. Strikingly, elderly patients with P-gp- de novo AML and favorable or intermediate cytogenetics had a CR rate of 81%. Patients with P-gp+ secondary AML with unfavorable cytogenetics had a CR rate of only 12%. Therefore, characterization of elderly AML patients at diagnosis using biologic parameters may help identify those patients who are likely to achieve a CR with conventional regimens, as well as those patients who require alternate treatment designed to overcome MDR. In contrast, expression of P-gp and functional drug efflux is detected in only 25% to 30% of AML patients less than 50 years of age. While P-gp expression is less strongly associated with a poor outcome in younger versus older AML patients, it remains strongly associated with resistant disease. Studies are ongoing to determine the prognostic significance of LRP and MRP in various forms of leukemia. PMID- 9408959 TI - Drug resistance in multiple myeloma. AB - The development of multidrug resistance (MDR) is a major obstacle to improving treatment outcomes in multiple myeloma. Recent studies have indicated that several specific mechanisms of MDR may be involved in clinically refractory multiple myeloma patients, such as expression of P-glycoprotein (P-gp), expression of the lung-resistance protein (LRP) and suppression of apoptosis via expression of Bcl-2. The emergence of these mechanisms of MDR in multiple myeloma is enhanced by exposure to chemotherapeutic agents. Recently, clinical reversal of MDR by noncytotoxic P-gp modulators such as verapamil, cyclosporin A (CsA), and PSC 833 was explored in acute leukemia and multiple myeloma. Preliminary results from clinical phase I/II trials indicate that reversal of MDR via modulation of P-gp is possible and that coadministration of these MDR modulators with chemotherapeutic agents alters the plasma pharmacokinetics of chemotherapeutic agents. Phase II and III clinical trials investigating the efficacy of these and other agents in the reversal of MDR in hematologic malignancies are ongoing. PMID- 9408960 TI - Pharmacologic approaches to reversing multidrug resistance. AB - The rationale for modulation of multidrug resistance (MDR) by inhibitors of the multidrug transporter, P-glycoprotein (P-gp) includes the following: (1) P-gp is expressed by human cancers, either at diagnosis or after failure of chemotherapy; (2) P-gp expression at diagnosis has been associated with a poor prognosis in some types of cancer; (3) MDR related to P-gp expression can be reversed by modulators, resulting in enhanced therapeutic efficacy in cellular and animal models of drug resistance; and (4) the emergence of MDR related to P-gp expression can be prevented in cellular models by co-administration of MDR related cytotoxins and modulators. Clinical trials of modulation of MDR have been limited by two major factors: inability to achieve adequate levels of the modulators to reverse drug resistance in patients and the presence of other mechanisms of resistance in tumor cells in addition to P-gp. The former limitation will hopefully be overcome by new, more potent and specific inhibitors of P-gp such as PSC 833. The latter will require further understanding of various alternative cellular mechanisms of resistance and the development of approaches to overcome or circumvent these mechanisms. PSC 833 is associated with significant drug interactions with MDR-related cytotoxic agents, which require dose reduction of the cytotoxins to achieve a dose exposure and toxicity similar to the chemotherapy agents without a modulator. These drug interactions are predictable and are at least in part due to inhibition of P-gp in normal tissues such as the liver and kidneys, where P-gp is known to play a role in drug excretion. Data from knockout mice, which lack P-gp expression, support the concept that P-gp is an important factor in MDR-related drug disposition. Early data from phase I and II trials with PSC 833 indicate that substantial inhibition of P-gp can be achieved in patients at clinically tolerable doses of both modulator and cytotoxins. The ultimate therapeutic benefit of MDR modulation with PSC 833 is currently being tested in phase III clinical trials in acute myeloid leukemias (AMLs) and multiple myeloma. PMID- 9408961 TI - Drug resistance to DNA topoisomerase I and II inhibitors in human leukemia, lymphoma, and multiple myeloma. AB - Several antineoplastic agents used in the treatment of hematologic malignancies exert their cytotoxic effects by inhibiting the activity of nuclear DNA topoisomerase (topo) I or II. Mechanisms of drug resistance to topoisomerase inhibitors have been defined at the molecular level from in vitro studies using model cell lines, and include quantitative and qualitative changes in topo I and II. The possible roles of these mechanisms in clinical drug resistance and clinical outcomes for patients with hematologic malignancies are now under investigation. Available data indicate that the blast content of topo II does not correlate with clinical outcome in acute myeloid leukemia (AML), and this may also be true in acute lymphocytic leukemia (ALL). Chronic lymphocytic leukemia (CLL) cells are resistant to topo II inhibitors because they express low levels of topo II. Further studies using sequential biopsy samples and assays of topoisomerase activity should establish the role that changes in topo I and II activity play in the development of drug resistance in hematologic malignancies. PMID- 9408962 TI - Measuring multidrug resistance expression in human malignancies: elaboration of consensus recommendations. AB - Before the prognostic significance of P-glycoprotein (P-gp) expression can be properly evaluated in prospective clinical trials of P-gp modulators, standard techniques for the measurement of P-gp must be widely accepted. Several multicenter trials have demonstrated large discrepancies in the observed levels of P-gp expression in the same clinical samples evaluated at different centers. The greatest discrepancies occurred with samples that expressed low levels of P gp. Although standardized procedures have dramatically increased the interlaboratory reproducibility of flow cytometry and polymerase chain reaction assays, data from immunocytochemistry remain difficult to interpret. Consensus recommendations are presented for improving data reproducibility. These recommendations emphasize the importance of using calibrated batches of antibodies and two different antibodies for immunocytochemistry, the need for an internal standard for reverse transcriptase-polymerase chain reaction (RT-PCR) assays, the need for the presentation of data as a continuous variable, and the need for setting standard parameters for flow cytometry. It is also extremely important for the success of clinical trials that multiple techniques be employed to insure accurate measurement of P-gp expression. PMID- 9408963 TI - On the function of pit cells, the liver-specific natural killer cells. AB - Pit cells are liver-specific natural killer (NK) cells and belong to the group of sinusoidal cells, together with Kupffer, endothelial, and fat-storing cells. Pit cells are lymphoid cells containing specific granules, classifying them also as large granular lymphocytes (LGL). They probably originate from the bone marrow, circulate in the blood, and marginate in the liver, where they develop into pit cells by lowering their density and increasing the number of granules, which decrease in size. Pit cells remain in the liver about 2 weeks and are dependent on Kupffer cells. Pit cells also proliferate locally, when stimulated with interleukin-2, biological response modifiers, or other agents. Pit cells adhere to tumor target cells during killing. They possess a high level of natural cytotoxicity against a variety of tumor cell lines, which is comparable to the cytotoxicity level of lymphokine-activated killer (LAK) cells. Tumor cell killing is synergistically enhanced when pit cells attack tumor cells together with Kupffer cells. Further investigations are needed to clarify the mechanisms of pit cell cytotoxicity and the role of these cells in killing virus-infected cells, such as during viral hepatitis. PMID- 9408964 TI - Interferons in host defense. AB - The Type I interferons are a group of related glycoproteins that play a key role in host defenses against viral infections. The interferons bind to a cell surface receptor and initiate the transcription of a wide range of proteins that have potent antiviral properties. The mechanism by which interferon binding to the cell surface initiates gene transcription has recently been elucidated and involves activation of protein kinases (JAK 1 and Tyk 2) followed by phosphorylation and activation of transcriptional regulators (the STAT proteins). These signal transduction molecules are not unique to the interferon signaling pathway, and other cytokines as diverse as erythropoietin and IL-2 use the same, or related proteins. To overcome the antiviral effects of the type I interferons, some viruses that cause chronic infections have developed interferon inhibitors that reduce the effectiveness of endogenous and exogenous interferon. PMID- 9408965 TI - Complement and complement deficiencies. AB - The complement system provides a first line of defense and mediates a large variety of cellular and humoral interactions within the immune response, including chemotaxis, phagocytosis, cell adhesion, and B-cell differentiation. The system involves more than 30 serum components and numerous cell surface regulators and receptors. Similar to the blood clotting system, complement activation is initiated through a series of complex activation cascades involving enzymatic cleavage. Three independent complement activation cascades, the classical, the alternative, and the lectin pathway, have been described. The liver is the main site of biosynthesis for most of the serum components of complement and diseases of the liver can lead to alterations of the normally stable plasma levels of complement. Deficiencies of single components can lead to a broad variety of secondary diseases, caused by either imbalanced activation or defects in the humoral or cellular response to microbial infections. PMID- 9408966 TI - The role of collectins in host defense. AB - Mannose-binding protein (MBP) belongs to a group of Ca(2+)-dependent lectins called collectins that play a role in first-line host defense. It recognizes specific carbohydrate residues (mannose and N-acetylglucosamine) on the surface of microorganisms and promotes the killing of microbes either by acting directly as an opsonin or by activating the lectin complement pathway. The collagenlike domain of MBP is important for the binding of MBP to the collectin receptors expressed on different phagocytes, and for activation of complement. The binding of MBP to bacteria, viruses, and parasites has been demonstrated in vitro. Three major mutations have been found in exon 1 of the MBP gene, which encodes the collagenous domain of the protein. These mutations cause low levels of serum MBP and have been linked with lifelong risk of infection. The homozygotes for these mutations are especially susceptible to severe infections. PMID- 9408967 TI - Bystander activation by cytokines of intrahepatic T cells in chronic viral hepatitis. AB - Although immune responses to hepatitis viruses are initiated by virus-specific T cells, there is evidence that many more intrahepatic T cells are activated than those specific for the pathogen. Recent evidence suggests that cytokine combinations, such as IL-2, IL-6, and TNF alpha, can activate both naive and memory T cells in vitro. The inflammatory cytokine milieu in the liver of patients with chronic viral hepatitis may therefore favor bystander activation of T cells. This may play an important role in enhancing effector T-cell function in the liver, and in maintaining peripheral memory T cells in the absence of antigenic stimulation, such as after virus clearance. PMID- 9408968 TI - Bacterial infections in liver disease. AB - Most bacterial infections in cirrhotic patients are hospital-acquired. Urinary tract infections, spontaneous bacterial peritonitis (SBP), respiratory tract infections, and bacteremia are the most frequent bacterial infectious complications seen in cirrhotic patients. SBP is the most characteristic infectious complication of cirrhotic patients, and it is defined as the infection of a previously sterile ascitic fluid, with no apparent intra-abdominal source of infection. The incidence of SBP in cirrhotic patients admitted to hospital with ascites has been estimated to range between 7 and 23%. The diagnosis is established on the basis of clinical signs and symptoms and/or a polymorphonuclear cell count in ascitic fluid higher than 250/mm3. This diagnosis is confirmed by a positive culture in approximately 70% of the cases. The remaining 30% are considered culture-negative SBP but are empirically treated with antibiotics because severe peritonitis and death may follow if these patients are not treated. Early diagnosis, the routine use of diagnostic paracentesis in patients admitted to hospital with ascites, and, especially, the use of adequate antibiotics are very important tools in the treatment of SBP. Third-generation cephalosporins are the first-choice antibiotic treatment in SBP, although selected patients with SBP, those with normal renal function and without hepatic encephalopathy, shock, or gastrointestinal bleeding, may be treated with oral quinolones. Selective intestinal decontamination with norfloxacin is safe and useful in the primary and secondary prophylaxis of SBP, although the incidence of quinolone-resistant organisms is increasing and this may be a problem in the future. PMID- 9408969 TI - The liver in AIDS. AB - Deficits in cell-mediated immunity in AIDS result in a wide variety of hepatic complications, including granulomas, cytomegalovirus hepatitis, multimicrobial AIDS cholangiopathy, Kaposi's sarcoma, and lymphoma. Kupffer cells are the major hepatic target cell population for human immunodeficiency virus-1 (HIV-1), and rhesus monkeys with simian immunodeficiency virus infection have served as a model for ultrastructural analysis of viral clearance by these cells. The majority of patients with established AIDS reveal abnormalities on serum liver tests. In these individuals, the differential diagnosis includes opportunistic infections and neoplasms, as well as possible concomitant chronic viral hepatitis B, C, D, and G, and drug hepatotoxicity. This article reviews the spectrum of hepatic pathology in AIDS and discusses the effects of HIV-1 infection on hepatitis virus infections. PMID- 9408970 TI - Host factors in chronic viral hepatitis. AB - The clinical outcomes of both hepatitis B and C virus infection are immensely variable, ranging from subclinical, self-limiting infection to end-stage liver disease with hepatocellular carcinoma. Knowledge of the host factors that determine these outcomes is important for the understanding and management of these diseases and may in the future guide rational drug development. Epidemiologic studies have elucidated the role of age (at the time of infection) and sex on disease outcome and the complex role of HIV coinfection has become clearer with time. More recently, investigation of genetic susceptibility to the most adverse outcomes of infection has identified the importance of polymorphisms in the MHC class I and II loci, mannose-binding protein, and the TNF alpha promoter. However, relative to malaria, the study of genetic susceptibility in viral hepatitis is still in its infancy. PMID- 9408971 TI - Liver tumors and host defense. AB - Complex molecular and cellular mechanisms exist to protect cells against tumor formation and to protect the entire organism against further development and spread of established tumors. The p53 tumor suppressor gene controls the cell cycle through at least two mechanisms, namely, mitotic arrest and apoptosis. Human hepatocellular carcinomas (HCCs) are often found to have mutant p53, or sometimes may have dysfunctional p53 as a result of its being bound by viral or cellular proteins. Another mechanism of host response is the production of transforming growth factor beta 1, which acts on receptors in normal hepatocytes to cause inhibition of DNA synthesis; abnormalities of transforming growth factor beta 1 have been documented in HCCs, but their biologic significance is unclear. Other host defense mechanisms include cellular responses to the tumor and the proliferation of substances with anticoagulant properties. PMID- 9408972 TI - World-wide surveillance of influenza. AB - Influenza epidemics and pandemics resulting in excess mortality are due to various influenza viruses, in which through the accumulation of mutations the structure changes. A world-wide surveillance has been set up for early detection of new influenza virus strains and of epidemics or pandemics resulting thereof. Basing on such data the World Health Organisation (WHO) issues recommendations to Public Health authorities on the most efficient means for prevention. PMID- 9408973 TI - Effect of retinoic acid on otic capsule chondrogenesis in high-density culture suggests disruption of epithelial-mesenchymal interactions. AB - Previous studies have shown that in utero exposure of the mouse embryo to nonphysiological levels of all-trans retinoic acid (RA) produces malformations of the epithelial-derived auditory and vestibular receptors of the inner ear and its surrounding cartilaginous capsule. In this study, we demonstrate the effects of all-trans RA in high-density cultures of the periotic mesenchyme fated to form the otic capsule. Our results demonstrate an inhibition of chondrogenesis in cultured periotic mesenchyme + otic epithelium of embryonic age E10.5 days (E10.5) in response to all-trans RA exposure. However, at later stages of development (i.e., E12, E14), when epithelial-mesenchymal interactions are no longer required for initiation of chondrogenesis, exposure to this teratogen has no effect on the chondrogenic process. Two analogues of all-trans RA, i.e., cis RA and trans-retinol, were investigated for their biological activity in chondrogenic cultures of inner ear mesenchyme + epithelium. Moreover, we tested the inductive capability and responsiveness of in utero RA-exposed inner ear tissues when cultured with inner ear tissues that were not exposed to this teratogen. Our results support the hypothesis that all-trans RA disrupts otic capsule formation by interfering with the tissue interactions required for its normal differentiation and development. PMID- 9408974 TI - Prenatal exposure to salicylates and gastroschisis: a case-control study. AB - Gastroschisis, which is a defect in the abdominal wall, lateral to the umbilical cord, is considered to be a vascular problem, probably due to a disruption of the omphalomesenteric artery [Hoyme et al. (1981) J. Pediatr. 98:228-231]. Recently, Torfs et al. [(1996) Teratology 54:84-92] observed a significantly increased risk for aspirin and ibuprofen, two strong cyclooxygenase inhibitors. Here we present the results of a case-control study conducted by the Spanish Collaborative Study of Congenital Malformations (ECEMC) on the relationship between prenatal exposure to salicylates during the first trimester of pregnancy and gastroschisis. The results show an increased risk (OR = 3.47; P = 0.015) after controlling the possible effect of maternal age and maternal smoking during pregnancy. PMID- 9408975 TI - Goldenhar syndrome among infants born in military hospitals to Gulf War veterans. AB - Reports in the popular press described the occurrence of Goldenhar syndrome among children of Persian Gulf War veterans (GWVs). The objective of this investigation was to compare the birth prevalence of Goldenhar syndrome among infants born in military hospitals to GWVs and to military personnel who were not deployed to the Gulf War (NDVs). Computerized hospital discharge data were reviewed for infants conceived after the war and born prior to the 1st of October, 1993, in medical treatment facilities (MTFs) operated by the U.S. Department of Defense. Medical records were evaluated for infants diagnosed at birth with at least one abnormality that might be related to Goldenhar syndrome. Two pediatricians, blinded to the parental Gulf War status of each infant, reviewed records. An estimated 75,414 infants were conceived after the Gulf War and born in MTFs during the study period (34,069 GWV infants and 41,345 NDV infants). Seven infants fulfilled the case criteria (five GWV infants and two NDV infants). All infants had fathers who served in the military at the time of their conception and birth. The birth prevalence was 14.7 per 100,000 live births among GWV infants (95% confidence interval [CI]: 5.4-36.4) and 4.8 per 100,000 live births (95% CI: 0.8-19.5) among NDV infants (relative risk: 3.03; 95% CI: 0.63-20.57; P values: [2-tailed] = 0.26, [1-tailed] = 0.16). The few affected cases and the broad confidence intervals surrounding the relative risk require that these results be interpreted with caution and do not exclude chance as an explanation for these findings. PMID- 9408976 TI - Critical period of carbon tetrachloride-induced pregnancy loss in Fischer-344 rats, with insights into the detection of resorption sites by ammonium sulfide staining. AB - Several low-molecular weight halocarbons have been shown to cause full-litter resorption (FLR), i.e., pregnancy loss, in Fischer-344 rats treated during organogenesis. To determine periods of gestation sensitive to acute exposure, a single dose of 150 mg carbon tetrachloride (CCl4)/kg was administered on gestation day (GD) 6, 7, 8, 10, or 12. Fetuses were delivered by cesarean section on GD 20. Non-gravid uteri were examined for resorption sites, placed in 10% ammonium sulfide, and re-examined for stained resorption sites approximately 1 and 4.5 hr later. FLR was seen in 4% (1/27) of control dams and 36% (4/11), 54% (7/13), 72% (18/25), 54% (7/13), and 0% (0/12) of dams treated on GD 6, 7, 8, 10, and 12, respectively. Ammonium sulfide staining clearly yielded a more accurate account of the incidence of FLR. The technique was most effective when the staining period was extended to 4.5 hr, as two cases of FLR were revealed that had been undetected after 1 hr of staining. For dams with FLR, staining was required to detect resorption sites in all dams treated on GD 6 or 7, most dams treated on GD 8, and one dam treated on GD 10. Fewer implantation sites were detected in the dams treated on GD 6, and the size of the stained resorption sites increased as the day of treatment was delayed. These findings demonstrate a relationship between the time of toxicant exposure and the size and detectability of resorption sites near term, suggesting that the size of the resorption site may reliably reflect the time of embryonic death. Treatment on GD 8 caused the highest incidence of FLR and will be used in subsequent mechanistic research. PMID- 9408977 TI - Pathogenesis of ectrodactyly in the Dactylaplasia mouse: aberrant cell death of the apical ectodermal ridge. AB - Dactylaplasia, or Dac, was recently mapped to the distal portion of mouse chromosome 19 and shown to be inherited as an autosomal semi-dominant trait characterized by missing central digital rays. The most common locus for human split hand split foot malformation, also typically characterized by missing central digital rays, is 10q25, a region of synteny to the Dac locus. The Dac mouse appears to be an ideal genotypic and phenotypic model for this human malformation syndrome. Several genes lie in this region of synteny, however, only Fibroblast Growth Factor 8, or Fgf-8, has been implicated to have a role in limb development. We demonstrate that the developmental mechanism underlying loss of central rays in Dac limbs is dramatic cell death of the apical ectodermal ridge, or AER. This cell death pattern is apparent in E10.5-11.5 Dac limb buds stained with the supravital dye Nile Blue Sulfate. We demonstrate that Fgf8 expression in wild type limbs colocalizes spatially and temporally with AER cell death in Dac limbs. Furthermore, in our mapping panel, there is an absence of recombinants between Fgf-8 and the Dac locus in 133 backcross progeny with a median linkage estimate of approximately 0.5 cM. Thus, our results demonstrate that cell death of the AER in Dac limbs silences the role of the AER as key regulator of limb outgrowth, and that Fgf-8 is a strong candidate for the cause of the Dac phenotype. PMID- 9408978 TI - Initiation of phenytoin teratogenesis: pharmacologically induced embryonic bradycardia and arrhythmia resulting in hypoxia and possible free radical damage at reoxygenation. AB - The aim of this study was to investigate if phenytoin has the capacity to induce embryonic hypoxia mediated via adverse effects on the embryonic heart. Mouse embryos of different strains (CD-1, C57B1/6J and A/J) as well as Sprague Dawley (SD) rat embryos were cultured in vitro (in 75-80% rat serum) by the whole embryo technique. Effects on the heart were examined on gestational day 10 for mouse embryos and days 11 and 13 for rat embryos. Phenytoin was dissolved in water to give concentrations of 50-800 microM. In the mouse embryo studies, phenytoin caused a concentration-dependent decrease in embryonic heart rate in all three strains, with a slight decrease at 100 microM (2-7%) and a more pronounced effect at 200 microM (approximately 20%). Temporary or permanent cardiac arrest occurred in 86% of the CD-1 embryos at 500 microM, in 67% of the C57B1/6JM at 400 microM, and in all A/J embryos at 300 microM. Arrhythmias was observed in 8% in CD-1 embryos at 200 microM, in 18% at 150 microM in C57B1/6J embryos, and in 67% of the A/J embryos at 100 microM (lowest tested concentrations where arrhythmias occurred). In rat embryos, a concentration-dependent decrease in heart rate was observed on both days 11 and 13 at similar concentrations as in the mouse embryo studies. In a separate experiment, the effects on the heart rate of free phenytoin (not serum protein bound) were examined in rat embryos cultured in serum-free medium. Already at 12 microM a significant decrease in heart rate was observed. Altogether, the results support the hypothesis that phenytoin teratogenicity is initiated by pharmacologically induced embryonic hypoxia. A genetic susceptibility to the adverse effects of phenytoin on the embryonic heart may be of importance to explain strain and species differences in phenytoin teratogenicity. PMID- 9408979 TI - Teratogen update: fetal effects of indomethacin administration during pregnancy. PMID- 9408980 TI - Transcranial Doppler for early identification of potential organ transplant donors. AB - BACKGROUND: Encouraging results in transplant medicine create a growing demand for organ transplant donors. Transcranial Doppler (TCD) has been used by several investigators to assess arrest of the cerebral circulation in brain dead patients. We report on TCD as a monitoring tool for early identification of potential organ transplant donors. DESIGN: A prospective clinical study. SETTING: Intensive care unit (ICU) of a 900-bed community hospital (primary and tertiary care center) in Vienna, Austria. SUBJECTS AND METHODS: All patients with acute intracranial lesions admitted to our intensive care unit underwent TCD examination at least once daily. In patients with Glasgow Coma Scores < 7, TCD waveforms with high resistance profiles unchanged by therapeutic attempts to lower intracranial pressure indicated the need for repeated TCD up to four times a day. TCD waveform abnormality consisting of absent or reversed diastolic flow or small early systolic spikes in at least two intracranial arteries was considered to constitute intracranial circulatory arrest. Brain death was confirmed by clinical criteria, an isoelectric electroencephalography (EEG) or non filling of the intracerebral arteries on arteriography. RESULTS: From January 1994 to July 1996 we identified 11 comatose patients as potential organ transplant donors with typical TCD findings indicating intracranial circulatory arrest. Diagnosis was subarachnoid hemorrhage in 7 and intracerebral hemorrhage in 4 patients. Brain death diagnosis according to the criteria of Austrian law was initiated immediately after the TCD findings suggested intracranial circulatory arrest. Confirmation of brain death was obtained by clinical criteria and either EEG (6 patients) or cerebral angiography (5 patients). CONCLUSION: TCD examinations on a daily routine basis offer a noninvasive monitoring method for early assessment of intracranial circulatory arrest. TCD enables quick identification and further diagnosis of candidates for organ transplant donation. PMID- 9408981 TI - Renal tolerability of four different once-daily dose regimen of netilmicin in critical care patients. AB - A prospective, randomized trial was conducted in a medical intensive care unit to assess safety and tolerability of four different dose regimens of intravenous netilmicin given once daily in the treatment of febrile episodes in critically ill patients. Eighty patients with febrile episodes during their stay in the intensive care unit were included in the study. The patients were randomized into four groups: Group 1 received a single daily dose of netilmicin based upon weight, age and renal function according to a dosage nomogram [13] (mean dose 298 +/- 29 mg, median 300 mg, range 250-350 mg), group 2 received 150% of this standard dose (mean 418 +/- 45 mg, median 400 mg, range 350-500 mg), group 3 200% (mean 525 +/- 41 mg, median 500 mg, range 400-550 mg) and group 4 250% (mean 710 +/- 39 mg, median 650 mg, range 600-750 mg). Duration of treatment was six days. Positive cultures were obtained in 29 patients. Serum creatinine and creatinine clearance, as well as netilmicin trough levels and levels of alpha 1 microglobulin showed no significant difference between the groups before, during, and after therapy. Our results indicate that with once daily dosing even high doses of netilmicin are well tolerated in patients with a creatinine clearance of > 70 ml/min before therapy. Necessary precautions include monitoring of drug trough levels (< 1 mg/L) and maintenance of adequate volume status. PMID- 9408982 TI - Primary adrenal insufficiency in two patients with the acquired immunodeficiency syndrome associated with disseminated cytomegaloviral infection. AB - We present two patients with manifest acquired immunodeficiency syndrome (AIDS) suffering from a generalized cytomegalovirus (CMV) infection. Over the course of several weeks they had developed a state of increasing lethargy and fatigue and one patient had noticed a darkening of his skin. These and other symptoms (vomiting, diarrhoea, hypotension) were suggestive of adrenal insufficiency. Laboratory findings included an increase of serum potassium levels, a decrease of serum sodium concentrations and elevated levels of the adrenocorticotropic hormone (ACTH). These findings, as well as the prompt therapeutic response to hydrocortisone established the diagnosis of adrenal insufficiency. Although definitive proof is lacking, generalised CMV infection is the most likely cause of our patients' symptoms. For the early initiation of appropriate substitution therapy, persons infected with the human immunodeficiency virus (HIV) with signs of CMV infection should be carefully and repeatedly monitored for clinical and laboratory signs of adrenal insufficiency. PMID- 9408983 TI - Ethics and nutrition. AB - Whilst much has been written on the implications of withholding or withdrawing nutritional support therapy from the terminally ill, the comatose or vegetative patients relatively little has been published on the ethical implications of nutritional therapy in other groups. This article outlines a surgeon's view of ethical issues as they apply to nutrition and concludes that failure to provide nutritional support in one form or another for patients who have or who are expected to have seven days or more of inadequate oral intake should now be considered unethical. PMID- 9408984 TI - Therapeutic results with Cerebrolysin in the treatment of dementia. AB - BACKGROUND: In the present study patients suffering from dementia were treated with an infusion therapy using the neurotrophic drug Cerebrolysin. The effectiveness of the therapy was examined under conditions of daily clinical practice. METHODS: 645 Patients were treated with 30 ml Cerebrolysin daily. The average period of treatment was 17.8 days. Prior to treatment patients were diagnosed according to DSM-III-R. In addition a differential diagnostic examination using the Hachinski Ischemic Score was performed. Clinical symptoms and the Clinical Global Impression (CGI) were determined before and after therapy. RESULTS: Cerebrolysin therapy led to a significant (p < 0.001) improvement in memory for 62% of the patients. 65% showed improvement in concentration, 50% in mood and fatigue, and 47% improved in vertigo. An improvement in the Clinical Global impression was observed in approximately 80% of the patients. The improvement of symptoms was significantly larger (p < 0.05) in younger, less afflicted patients than in the older, more seriously ill patients. The unusual good tolerance of Cerebrolysin was especially noteworthy. CONCLUSIONS: The present study confirms the results of earlier clinical studies with Cerebrolysin in the treatment of Alzheimer patients and patients with vascular dementia in which similar responder rates were observed. The significant dependence of therapeutic success on the length and severity of illness confirms that timely pharmacological intervention leads to best therapeutic results. PMID- 9408985 TI - Protein analysis by packed capillary liquid chromatography with electrospray ionization and time-of-flight mass spectrometry detection. AB - Packed capillary liquid chromatography (LC) has been interfaced on-line to electrospray (ES) ionization with time-of-flight (TOF) mass spectrometry (MS) detection for the separation and identification of standard proteins. Using this system, as little as 2.5 fmol of cytochrome c could be detected with a 0.1 mm I.D. column. The effect of column size on sensitivity has been investigated using various column diameters and a sensitivity increase of 428-fold has been observed between the 0.10- and 2.1 mm I.D. columns. Although the TOF-MS can ultimately record data at up to 100 mass spectra/s, the peak widths for these particular separations require only 1 spectra/s. Finally, the sensitivity obtained is a function of both the reduced diameter LC column and the high ion duty cycle of the TOF-MS. PMID- 9408986 TI - Polychlorinated naphthalenes in groundwater samples from the Llobregat aquifer (Spain). AB - Polychlorinated naphthalenes (PCNs) have characteristics fairly similar to those of polychlorinated biphenyls and terphenyls (PCBs and PCTs) and although they have been used in numerous applications, very few reports on the occurrence of PCNs in water have been published. Both automated solid-phase extraction (SPE) and liquid-liquid extraction (LLE) procedures were evaluated to concentrate PCNs. High-resolution gas chromatography with electron-capture detection (HRGC-ECD) and mass spectrometric detection in the selected ion monitoring mode (SIM) were used to analyze PCNs in groundwater samples from the aquifer of the Llobregat river near Barcelona (NE Spain). The homologue distribution profiles of PCNs found in groundwater samples and the standard mixtures of Halowax (1099, 1013 and 1014) were compared. Quantification was achieved by HRGC-ECD. Total PCN concentrations expressed as Halowax 1099 equivalents ranged from < 0.5 ng/l to 79.1 micrograms/l in the water samples, with tetrachloronaphthalenes (TetraPCN) as the major group of congeners. PMID- 9408987 TI - Optimized method for the determination of organophosphorus pesticides in meat and fatty matrices. AB - A method was developed and optimized for determination of residues of organophosphorous pesticides (OPs) in meat and fatty matrices. The method was developed for the national Danish monitoring programme, whereby priority was given to simplify the clean-up, avoid use of toxic and harmful organic solvents and allow quantification at ppb level using GC with nitrogen-phosphorus detection (NPD). Homogenized meat was extracted using ethyl acetate, the co-extracted water being removed using anhydrous Na2SO4. The clean-up was done through fat precipitation by cooling the extract flowed by solid-phase extraction on C18 mini columns. Pesticide residues were determined by GC-NPD using a DB1701 capillary column. The limit of detection was 1 to 20 ppb and limit of determination was 2 to 33 ppb and the method is feasible for control of fat-soluble OPs according to the maximum residue limits set by the European Communities. The method developed covers a broad polarity range from polar OPs, such as acephate and methamidophos, through medium polar OPs to non-polar pesticides, such as prothiofos. PMID- 9408988 TI - Selectivity of electron-donor- and electron-acceptor-bonded silica packing materials for hydrophobic environmental contaminants in polar and non-polar eluents. AB - Electron-acceptor-bonded stationary phases, 2-(nitrophenyl)ethylsilyl (NPE) and 3 (p-nitrophenoxy)propylsilyl (NPO), and electron-donor-bonded phases, 3-(N carbazolyl)propylsilyl (CZP), 2-(1-pyrenyl)ethylsilyl (PYE), and 5 coronenylpentylsilyl (COP), were prepared from silica particles and their selectivities were examined in both polar and non-polar solvents for specific isomers of polychlorodibenzo-p-dioxins (PCDDs), hexachloronaphthalenes (HxCNs) and planar and non-planar polychlorobiphenyl (PCB) congeners. Although no single stationary phase was able to separate all the isomer pairs that are coproduced during the synthesis of the PCDDs and HxCNs, pairs can be separated by selecting a suitable stationary phase and solvent. The separation of mixtures of PCDD isomers were found to be most successful with PYE and NPO phases, which yielded the opposite elution orders for each isomer pair that is produced as a mixture. Similar results were obtained for the HxCN isomers that were separated on PYE and CZP phases. The COP phase provided easier separation of non-ortho-substituted and mono-ortho-substituted PCBs from the other PCBs based on the planarity than PYE phase. PMID- 9408989 TI - Evaluation of two solid-phase extraction procedures for the preconcentration of chlorophenols in drinking water. AB - Two off-line concentration procedures for the determination of sixteen chlorophenols in drinking water were developed. One involves acetylation of the samples and their subsequent preconcentration over graphitized carbon black cartridges. In the other, chlorophenols are derivatized following preconcentration over cross-linked styrene-divinylbenzene. The two proposed procedures are compared in terms of chlorophenol recoveries, throughput and breakthrough volume of the cartridges. The acetylated derivatives of chlorophenols are determined highly selectively at the concentration levels established by international legislation using gas chromatography in combination with microwave induced plasma atomic emission spectroscopy. PMID- 9408990 TI - Analysis of 2,4,6-trichloroanisole in wines using solid-phase microextraction coupled to gas chromatography-mass spectrometry. AB - Cork taint in wine is a serious problem which is exacerbated by the difficulty of its assessment. Current analytical procedures are costly, time consuming and require the use of large amounts of solvents. We developed and evaluated a rapid method for the detection of the cork taint compound, 2,4,6-trichloroanisole (TCA), in wine samples. The method employs solid-phase microextraction, a solventless, automated sampling procedure, coupled to GC-MS-selected ion monitoring analysis. Quantification is enabled by a fully deuterated [2H5]TCA analog used as an internal standard. Accuracy (+/- 8%), precision (R.S.D. 5-13%), and limit of quantification (5 ng/l) are comparable to existing methods. PMID- 9408991 TI - Enantiomeric determination of amino compounds with high sensitivity using the chiral reagents (+)- and (-)-1-(9-anthryl)-2-propyl chloroformate. AB - New chiral precolumn reagents, (+)- and (-)-1-(9-anthryl)-2-propyl chloroformate (APOC), are introduced for the chiral separation of amino acids and small peptides in capillary electrophoresis. Chiral separation of 17 amino acids and four small peptides as their diastereomeric 1-(9-anthryl)-2-propyl carbamate derivatives have been achieved by micellar electrokinetic chromatography. The detection limit for the derivatives is in the femtomole range with UV detection and in the attomole range with laser-induced fluorescence (LIF) detection. LIF detection was used to determine the enantiomeric excess of four APOC-derivatised peptides. The use of the new, anthracene-based reagents in conjunction with argon ion LIF makes enantiomeric determinations at ppm levels feasible. In this paper determinations below promille levels are performed without overloading the separation system. PMID- 9408992 TI - Displacement chromatography with on-column isomerization. AB - Displacement chromatography was simulated for the separation of two feed components interconverting by a reversible first order reaction and with Langmuirian adsorption behavior. The study was prompted by recent interest in the isolation of cis and trans forms of peptides containing one or more peptidyl proline residues when the isomerization reaction interferes with the separation. The parameter values used in the simulations are similar to those found experimentally by reversed-phase chromatography and capillary electrophoresis of phenylalanine-proline dipeptide. From the concentration profiles computed by the finite difference scheme, the dependence of both the yield and production rate on the temperature, column length, flow velocity and displacer concentration was evaluated. The most important operational variable of the system is the temperature as it affects both the kinetic and adsorption parameters. The yield and production rate of the component of interest were evaluated as a function of the column length and displacer concentration under conditions that facilitate its efficient separation and the plots show an optimum. Nonetheless, optimal conditions for yield and production rate were considerably different. In the temperature range from 2 to 42 degrees C, the yield always decreases with increasing temperatures and for all the cases, optimum yield by displacement mandates the use of conditions such as pH, solvent and temperature under which the rate of interconversion is reduced to a level where it does not palpably interfere with the separation. On the other hand, under certain conditions optimal production rate can be obtained at higher temperatures. PMID- 9408993 TI - Theoretical concepts of on-line liquid chromatographic-biochemical detection systems. I. Detection systems based on labelled ligands. AB - A theoretical foundation for on-line coupling of liquid chromatography (LC) with fluororeceptor assays based on fluorescent ligands is presented. Using a recently developed LC-receptor affinity detection (RAD) system as a model, equations are derived which describe the detector response and the signal-to-noise ratio as a function of important biochemical and instrumental parameters. The effect of ligand and label affinity, and receptor concentrations on the detector performance were investigated. It was found that the response of the RAD system is correlated with the affinity of the injected compounds and that the relative affinity order of binding ligands is maintained in the RAD system. PMID- 9408994 TI - Ligand structure of the divinylsulfone-based T-gel. AB - The requirements for divinylsulfone (DVS)-based gels to act as thiophilic adsorbents, binding immunoglobulins in a salt-dependent manner have been examined. No differences in protein binding were observed for a DVS-activated gel reacted with mercaptoethanol (the T-gel), or for the same gel treated at high pH to hydrolyse the active groups and/or allow the formation of cross-links within the matrix, indicating that an O atom may be substituted for the thioether without affecting the thiophilic interactions. Extending the time of the activation reaction between DVS and the matrix results in increased amounts of sulfone attached to the gel, but decreased levels of active vinyl groups. When coupled to mercaptoethanol, these adsorbents bound more IgG than gels activated for shorter periods. This provides a convenient method to prepare thiophilic adsorbents of high capacity while minimising the amount of DVS used. The immobilised vinylsulfone must be linked to an electron donating atom for IgG to bind. When the vinyl was instead reduced with sodium borohydride, protein binding was decreased. No IgG bound to amine-coupled DVS-activated adsorbents, perhaps due to an overall positive charge on these gels at pH 7.4. The binding of human IgG to the adsorbents is dependent on ligand density, with little protein binding to gels having less than 16 mumol sulfone per ml. The binding increased with the ligand density above this level, with more than 25 mg IgG binding per ml to an adsorbent having 114 mumol sulfone per ml. The lack of binding at low ligand densities would be expected if the IgG must interact with two or more sulfone ligands to be retained on the adsorbent. PMID- 9408995 TI - Liquid chromatography-atmospheric pressure ionization mass spectrometry for the determination of chloro- and nitrophenolic compounds in tap water and sea water. AB - Liquid chromatography coupled to atmospheric-pressure ionization mass spectrometry (LC-API-MS) with negative ion detection was studied for the determination of a variety of phenolic compounds in environmental waters. An isocratic mobile phase of 0.05% acetic acid-acetonitrile (50:50, v/v) was used. The influence of post-column addition of different bases on the sensitivity of the detection in electrospray (ES) was studied. The [M-H]-ion was the base peak for all the compounds using both ES and atmospheric-pressure chemical-ionization (APCI) ion sources. Moreover, abundant structural information was obtained by increasing the extraction voltage. Detection limits for standard solutions ranging from 2 to 13 ng injected for LC-ES-MS and from 0.02 to 20 ng for LC-APCI MS were obtained. Good reproducibilities (day-to-day and run-to-run) were observed. The optimum LC-ES-MS and LC-APCI-MS conditions thus determined were used for a quantitative analysis of some phenolic compounds in spiked tap water and sea water samples. PMID- 9408996 TI - Selection of optimum affinity tags from a phage-displayed peptide library. Application to immobilized copper(II) affinity chromatography. AB - Immobilized metal affinity chromatography (IMAC) is a versatile tool for the purification of proteins with affinity for immobilized metals. Moreover, this technique has also been used for the separation of proteins that do not exhibit significant metal affinity in the native form, by their fusion to a short metal binding peptide (a tail), most commonly, a sequence consisting of six adjacent histidine residues (His6). A phage-displayed random hexamer library is used to select for peptides with affinity for immobilized copper. The study follows our previous investigation in which a stringent selection protocol led to the selection of only one copper-binding peptide containing two histidines. The less stringent conditions employed in this work resulted in the selection of a more diverse population of peptides, but again, dominated by peptides containing two histidines (13 out of 19). The prevalence of peptides with two histidines, in contrast to peptides with a higher number of histidines (e.g. His6 or HHHMVH), is explained based on the differences in the pH dependence of their affinity for copper. As discussed, the selected peptides with two histidines will be superior affinity tails than peptides with a higher histidine content (e.g. His6). Moreover, a peptide with a single histidine but with a very high copper affinity, is also identified. Its high copper affinity is related to the presence of several hydrophobic residues in the neighborhood of histidine. Chromatography of human interleukin-1 beta (hIL-1 beta) and several other proteins containing a single surface-exposed histidine surrounded by several hydrophobic residues confirmed that such a sequence could also serve as a very effective metal binding domain for protein purification using immobilized copper(II) columns. PMID- 9408997 TI - Characterization of antigen-antibody complexes by size-exclusion chromatography coupled with low-angle light-scattering photometry and viscometry. AB - In this paper, the molecular masses (M(r)s) of the complexes of monoclonal anti BSA (antibody to bovine serum albumin) (clone: 33) and monomer BSA were determined on-line by using size-exclusion chromatography (SEC) coupled with a low-angle laser light-scattering (LALLS) detector and two concentration detectors, ultraviolet (UV) and refractive index (RI) (SEC-LALLS/UV/RI system). Also, the size and M(r)s of the complexes were evaluated by the SEC LALLS/UV/viscometer (VISC) system. This study demonstrated that, for small size macromolecules, the combination of light scattering and viscosity detection was a suitable choice for determining their M(r)s and sizes. PMID- 9408998 TI - Concurrent liquid chromatographic separation and photodiode array detection of retinol, tocopherols, all-trans-alpha-carotene, all-trans-beta-carotene and the mono-cis isomers of beta-carotene in extracts of human plasma. AB - In this report we describe the development of a method for the concurrent reversed-phase high-performance liquid chromatographic separation and photodiode array detection of human plasma retinol, tocopherols and carotenes. For a single sample injection, retinol, retinyl acetate, alpha-tocopherol, gamma-tocopherol, alpha-tocopheryl acetate, all-trans-alpha-carotene and all-trans-beta-carotene, as well as the mono-cis geometrical isomers of beta-carotene were separated and detected. Analytical separations were performed at a subambient temperature (0 degree C) over a Suplex pKb-100 reversed-phase analytical column with an isocratic mobile phase of methanol-methyl tert.-butyl ether-water (80:20:5, v/v/v) at a flow-rate of 0.8 ml/min for 60 min. Standards and samples were reconstituted in ethanol, and typically, 50 microliters was injected for analysis. By HPLC, compounds of interest were clearly resolved and detectable at the picomole level. PMID- 9408999 TI - High-performance liquid chromatographic determination of benomyl and carbendazim residues in apiarian samples. AB - Simple procedures for the extraction and chromatographic determination of benomyl and carbendazim in honey, bees wax, larvae, bees and pollen are proposed. The fungicides were extracted from honey, larvae and bees using ethyl acetate, while methanol was more suitable for wax and pollen samples. Pollen extracts need a further clean-up step with n-hexane. The determination is carried out by reversed phase high-performance liquid chromatography (HPLC) with fluorescence detection. The procedures have been applied to the analysis of benomyl on honey and larvae samples from hives whose bees were nourished with artificial food mixed with benomyl. PMID- 9409000 TI - Sensitive and selective determination of methylenedioxylated amphetamines by high performance liquid chromatography with fluorimetric detection. AB - A rapid, sensitive and selective liquid chromatographic method with fluorimetric detection was developed for the separation and quantification of four methylenedioxylated amphetamines without interference of other drugs of abuse and common substances found in illicit tablets. The method was validated by examining linearity, precision and accuracy as well as detection and quantification limits. Methylenedioxylated amphetamines were quantified in eight tablets from illicit drug seizures and results were quantitatively compared to HPLC-UV analyses. To demonstrate the better sensitivity of the fluorimetric detection, methylenedioxylated amphetamines were analyzed in serum after a liquid-liquid extraction procedure and results were also compared to HPLC-UV analyses. PMID- 9409001 TI - Detection of glycosaminoglycans as a copper(II) complex in high-performance liquid chromatography. AB - Glycosaminoglycans including heparin, heparan sulfate, chondroitin sulfate, dermatan sulfate and hyaluronan were analyzed by high-performance gel filtration chromatography. Detection was achieved at 240 nm based on the formation of copper(II) complex in copper sulfate solution at low pH. Detection of the copper(II)-heparin complex is sensitive, permitting the analysis of as little as 10(-7) g. This method was also successfully applied to the analysis of the chemically derivatized glycosaminoglycans (de-N-acetylated and/or oversulfated chondroitin sulfate) that cannot be depolymerized by enzymes such as heparin and chondroitin lyases. PMID- 9409002 TI - Determination of the insecticide imidacloprid in water and soil using high performance liquid chromatography. AB - We describe an analytical technique for measuring residues of imidacloprid, a relatively new and highly active insecticide, in water and soil using high performance liquid chromatography (HPLC). All analyses were performed on reversed phase HPLC with UV detection at 270 nm using a mobile phase of acetonitrile-water (20:80, v/v). Fortified water samples were extracted with either solid-phase extraction (SPE) or liquid-liquid extraction methods. A detection limit of 0.5 microgram/l was achieved using the SPE method. The imidacloprid residues in soils were extracted with acetonitrile-water (80:20, v/v), and the extract was then evaporated using a rotary evaporator. The concentrated extract was redissolved in 1 ml of acetonitrile-water (20:80, v/v) prior to analysis by reversed-phase HPLC. A detection limit of 5 micrograms/kg was obtained by this method which is suitable for analysis of environmental samples. Accuracy and precision at 10 and 25 micrograms/kg soil samples were 85 +/- 6% and 82 +/- 4%, respectively. PMID- 9409003 TI - Rapid determination of 1,4-dioxane in water by solid-phase extraction and gas chromatography-mass spectrometry. AB - A highly sensitive and accurate gas chromatography-mass spectrometry method for rapidly quantitative analysis of dioxane in water samples is described. Dioxane in water sample was extracted by hexane-methylene chloride (80:20, v/v) then transferred to a C18 solid-phase extraction (SPE) cartridge and eluted by acetonitrile. An aliquot of this solution was analyzed by gas chromatography-mass spectrometry with electron impact ionization and selected ion monitoring (dioxane m/z 88) mode. The quantitative limit was set to be 0.05 microgram/ml with an injection volume of 2 microliters and a sample volume of 1 ml. The isotopically labelled [2H8]dioxane (m/z 96) was used as internal standard. Analysis time is approximately 6 min. The results for various kinds of SPE cartridges in this method are also discussed. PMID- 9409004 TI - Identification of nonprotein amino acids from cycad seeds as N-ethoxycarbonyl ethyl ester derivatives by positive chemical-ionization gas chromatography-mass spectrometry. AB - Nonprotein amino acids from nine species of cycad seeds were analyzed as N ethoxycarbonyl ethyl ester (ECEE) derivatives by positive chemical-ionization gas chromatography-mass spectrometry. Based on the retention times and mass spectrometry analyses, 12 nonprotein amino acids were identified in these seeds. In addition to the excitatory and putative neurotoxin beta-N-methylamino-L alanine (BMAA), the known neurotoxin beta-N-oxalylamino-L-alanine (BOAA) was detected from the seeds of Macrozamia moorei and M. communis, and delta-N-oxalyl ornithine was obtained from the Cycas revoluta seeds. A novel nonprotein amino acid named cycasindene, previously reported from C. revoluta, was also found in the seeds of members of the C. angulata and C. rumphii complex. Eight additional known nonprotein amino acids were also identified. This is the first report of the neurotoxin BOAA from cycad seeds. PMID- 9409005 TI - Coupling chemical derivatization reactions with supercritical fluid extraction. AB - Coupling chemical derivatization reactions with supercritical fluid extraction for the determination of trace levels of organic and organometallic compounds in liquid and solid matrices is reviewed. Derivatization is used to increase the solubility of analytes in supercritical carbon dioxide, to increase analyte volatility for gas chromatographic analysis and to integrate sample preparation steps in order to reduce analysis time and costs. Reactions that are covered in this review derivatize analytes possessing carboxyl, hydroxyl, sulfonic and amino groups to their alkyl, acyl and silyl derivatives. Derivatization is also used to derivatize active matrix sites to facilitate the release of analytes. Approaches used to couple derivatization with supercritical fluid extraction include reactions conducted prior to extraction, under in-site supercritical fluid conditions, or off-line under injection port conditions. Examples of applications are given for organic and organometallic compounds in environmental, pharmaceutical and agricultural product samples. PMID- 9409006 TI - Gas chromatographic determination of organic acids from fruit juices by combined resin mediated methylation and extraction in supercritical carbon dioxide. AB - A procedure in which anionic analytes, trapped on ion exchange resin, are simultaneously methylated and released using methyl iodide in either supercritical carbon dioxide or acetonitrile has been extended to polyfunctional organic acids. The combined SFE methylation of fruit juice acids trapped onto ion exchange resin proceeds in good yield producing the methyl esters of fumaric, succinic, malic, tartaric, isocitric and citric acids which are readily separated by GC. Using this procedure low concentrations of one acid can be detected and quantitated in the presence of very high concentrations of another. This new method detects tartaric acid at levels of 10 ppm in juices containing 10,000 ppm citric acid. Quantitation was performed either by using GC-FID with triethyl citrate or diethyl tartrate as internal standards or with the element specific calibration capability of the GC-AED. A simple new technique for the determination of citric/isocitric acid ratio is now available. Also, in contrast to HPLC methods, the identity of an analyte is readily confirmed by GC-MS. PMID- 9409007 TI - Supercritical fluid extraction of pesticides in foods. AB - This article summarizes research findings involving the supercritical fluid extraction (SFE) of pesticides in food and other tissue matrices. Emphasis is placed on multiresidue analysis of pesticides in nonfatty foods, including some previously unpublished aspects of SFE in this application. Brief overviews of pesticides and traditional multiresidue methods are given, followed by discussion of results for SFE applications in the pesticide residue analysis of foods. PMID- 9409008 TI - Evaluation of a fibrous cellulose drying agent in supercritical fluid extraction and pressurized liquid extraction of diverse pesticides. AB - A fibrous cellulose powder (CF-1) was investigated as a drying agent for supercritical fluid extraction (SFE) and pressurized liquid extraction (PLE), also known as accelerated solvent extraction. Analysis of fifty-eight diverse pesticides was performed using gas chromatography-ion-trap mass spectrometric detection (GC-ITD). Extraction efficiencies were correlated versus pesticide polarity with samples of different water-CF-1 ratios. The effect of water was much more pronounced in SFE using CO2 than PLE using acetonitrile. Pesticide recoveries and limits of detection of fortified tomato samples mixed with CF-1 were determined. PLE gave recoveries > 80% for nearly all pesticides, and SFE gave similar recoveries except for the most polar and non-polar pesticides. SFE typically gave lower detection limits than PLE due to fewer matrix interferants. PMID- 9409009 TI - Sub- and supercritical fluid extraction of trichloropyridinol from soil prior to immunoassay. AB - A comparative study on the extraction of TCP (3,5,6-trichloro-2-pyridinol, a metabolite of chlorpyrifos) from soil with CO2 and H2O is reported. The polarity of the analyte requires the presence of both a cosolvent (methanol) and an ion pair reagent [(1R)-(-)-10-camphorsulfonic acid ammonium salt] for 95% extraction in 30 min when supercritical CO2 at 40 degrees C and 383 bar is used as extractant. Subcritical water (250 degrees C and 200 bar) enables complete extraction within 15 min without additives. Quantitation of the target analyte is performed by specific immunoassay using a non-commercial monoclonal antibody which provides a linear determination range between 0.005 and 5 micrograms/g, with coefficients of variation of 5.3 and 4.9% for the SC-CO2 and sub-H2O extractions, respectively. PMID- 9409010 TI - Development of a supercritical fluid extraction method for determination of lipid classes and total fat in meats and its comparison with conventional methods. AB - A method has been developed for the determination of total fat and lipid classes in meat using supercritical fluid extraction (SFE). The results agreed well with results from the conventional Bligh and Dyer and Schmid, Bondzynski and Ratzlaff extraction methods. SFE has advantages compared to the latter methods of a low consumption of hazardous organic solvents and shorter extraction time. After investigation of several different conditions, the most rapid extraction was achieved by adding 1 ml of cyclohexane to a 0.5-g sample mixed with 1 g Hydromatrix in a 7-ml thimble. The optimized SFE parameters were: 370 bar, 50 degrees C, 8% ethanol modifier, 4 ml/min dynamic flow for 30 min and collection with a tube leading to a vial. PMID- 9409011 TI - Extraction of artemisinin and artemisinic acid from Artemisia annua L. using supercritical carbon dioxide. AB - Artemisinin (an antimalaric compound) and its major precursor artemisinic acid, isolated as the active principles of the medicinal plant Artemisia annua L., were extracted by supercritical fluid extraction (SFE) and analyzed by supercritical fluid chromatography (SFC) using a capillary column, coupled with a flame ionization detector (FID). With optimized operating conditions, artemisinin and artemisinic acid were quantitatively extracted at a flow-rate of 2 ml min-1 in less than 20 min. The supercritical fluid was composed of carbon dioxide and 3% methanol with temperature and pressure fixed at 50 degrees C and 15 MPa, respectively. From the kinetic curves, it appears that the extraction of artemisinin is not limited by the diffusion of the analyte from the plant into the extraction fluid but rather by the elution process. These conditions avoided degradation of the analyte and gave clean extracts ready to be analyzed by SFC. The SFE-SFC-FID method was successfully applied to six samples of A. annua containing various concentrations of artemisinin and artemisinic acid. Results were compared with two conventional liquid solvent extraction processes. PMID- 9409013 TI - Beneficial effects of radiation and regulatory policy. AB - Adaptive and stimulating effects of ionizing radiation occur at near natural doses. This disagrees with linear, no-threshold hypothesis on the dose/effect relationship, which is a basis of the current radiation protection. Vast literature demonstrates that such effects, usually known as hormetic ones, occur at molecular, cellular and population levels, and often result in increase longevity and decreased cancer incidence. Exposure to lower than natural radiation causes deficiency symptoms in protozoa and bacteria. Hormetic effects suggest that the current radiation protection regulations may be too conservative. After the Chernobyl accident, adverse health effects and vast material losses were induced in the former USSR by practical implementation of the ICRP radiation protection recommendations. A revision of the current approach to managing the risks of ionizing radiation is needed for the public interest. PMID- 9409014 TI - Dosimetry of beta emitting radionuclides for use in balloon angioplasty. AB - The dose at varying distances from the surface of an infinite cylinder containing 90Y, 32P and 188Re respectively is calculated using published scaled point dose kernels for these three radionuclides. It is shown that all are suitable radionuclides for use in the irradiation of arteries subsequent to balloon angioplasty. All three may be used as a radioactive liquid in the angioplasty balloon, thereby simplifying the procedure and enabling a uniform radiation dose to be given to the arterial wall. It is however shown that there is a rapid reduction in dose with distance from the arterial wall which demands careful specification of the prescribed radiation dose. A similar rapid reduction with distance is also found with a central radioactive wire or with a radioactive stent containing the same radionuclides. PMID- 9409015 TI - Neural networks for signal processing applications: ECG classification. AB - QRS complexes were classified using a Multi Layer Perceptron (MLP) using a modified version of the original Backpropagation (BP) algorithm. The input to the network were bitmaps of the QRS complexes represented in the form of a 20 x 20 matrix. A number of hidden layers (and neurons in each hidden layer) were experimented upon to observe the rate at which the network converged. Larger Networks were observed to find the minimum on the error curve with ease. Increasing the network size beyond a certain size did not improve the performance rate, rather it decreased the performance rate. It is evident that there exists an optimal neural network architecture for every given problem. The weights change rules in Backpropagation algorithm were modified to include a variation of the relationship between momentum and learning rate to observe any increase in network's performance rate. A learning rate adaptation factor was introduced into the learning algorithm to decrease the network's chances of missing a minimum on the error curve. The network was found to perform extremely well with the modified version of the algorithm. The network converged after only 9000 learning cycles when compared to 14,000 cycles with the original algorithm. PMID- 9409016 TI - A computational model for blood flow through highly curved arteries with asymmetric stenoses. AB - In this paper, we develop a three dimensional model of blood flow through curved arteries with asymmetric stenoses. Firstly, the Navier-Stokes equations representing Newtonian flow are solved using PHOENICS, a computational fluid dynamics package which utilises the Finite Volume method of solution. The severity of the stenoses considered in this study vary from about 40% to about 70%. Subsequently, the model is solved for Reynolds numbers ranging from 100 to 1200. The pressure drop results show good agreement with published results. The results also show that stenoses on a bent artery has a significant effect on blood flow characteristics. PMID- 9409017 TI - Optimisation in radiotherapy I: defining the problem. AB - Optimisation in radiotherapy should incorporate a very wide set of variables, including the combinations of dose due to external beam radiotherapy, brachytherapy, internally administered radionuclides and the effects of chemotherapy, surgery, hyperthermia, other biological and chemical defences, alternative treatment techniques, lifestyle and mental state of the patient, the economics of cancer treatment, and consideration of tolerable levels of adverse effects and palliation. A full treatment optimisation would consider the influence and covariance of all these variables and any future techniques, as well as the complex constraints imposed by the biological systems being irradiated. This series of reviews concentrates on optimisation of radiotherapy through the treatment planning component of the treatment process--an area of radiotherapy research that has received a great deal of attention as the attached lists of references will testify. It hopes to provide the medical physics and engineering community (and hopefully the clinical community) with a background into the mathematical bases for the manipulation of radiation for clinical benefit. It also examines the potential benefits of research into these techniques in the light of recent approaches to optimisation in radiotherapy, and provides pointers to more concise accounts in the literature. In this first article, the incentive for radiotherapy optimisation research is established, and the actual radiotherapy optimisation problem (in terms of the manipulation of degrees of freedom in radiation delivery) is defined. The degrees of freedom associated with radiotherapy treatment are identified, and it is shown how these degrees of freedom translate into the mathematical parameters of the problem, including the dose distributions they produce. The constraints and objectives of the problem are also discussed from both physical and radiobiological perspectives. Subsequent articles will consider algorithms for performing the optimisation, their relative utility, and their application in clinical practice. PMID- 9409018 TI - Off-axis output factors for 6MV and 18MV photons. AB - A series of in-air measurements showed that collimator scatter (Sc) did not change significantly for 6MV photons when the centre of the field was moved away from the central axis. This result enabled a model to be developed for the off axis Effective Output Factor (EOF) which was then verified for 6MV and 18MV photons on a Varian 2100c accelerator and for 6MV photons on a Varian 600c accelerator. Thus off-axis output factors may be predicted, for a range of rectangular asymmetric fields, using only the Primary Off-Centre Ratio (POCR) in air and the on-axis output factor. Depth doses were also investigated off-axis and found to have no clinically significant differences compared with on-axis depth doses, for depths less than 7.5 cm for 6MV and 12.5 cm for 18MV photons. The model is simple to implement and avoids the need for a measurement for each patient, thus saving accelerator time. PMID- 9409019 TI - Conversion of an infrared densitometer for radiochromic film analysis. AB - By the simple incorporation of a high intensity red LED into a typical infrared film densitometer, radiochromic film can be analysed using existing detectors and scanning software. Results show an accurate dose measurement using radiochromic film and this system compared to conventional detectors for percentage depth dose and penumbral measurements in high and low energy x-ray beams. A small circuit including a red Light Emitting Diode (LED) was positioned inside the film densitometer which does not obscure the infrared source. The red and infrared diodes work independently. For 6 MV x-rays, the 80%/20% penumbral width at 15 mm depth for a 10 x 10 cm field at 100 cm SSD was measured to be 3.5 mm with radiochromic film as compared to 3.3 with corrected diode measurements. Percentage depth doses were measured to within +/- 3% of ionisation chamber data at 6MV and within +/- 2% for 250 kVp x-ray with the film placed parallel to the beam direction in both cases. PMID- 9409020 TI - Determination of correct AEC function with computed radiography cassettes. AB - It has long been recognized that the assessment of Automatic Exposure Control (AEC) function is an important criterion in the assessment of any general radiographic x-ray installation. Of the tests commonly performed on an AEC, only Beam Quality Dependence relates intimately with the optical density produced on the resultant radiograph. In this test the AEC function is ascertained as being satisfactory or otherwise, on the basis of whether the optical density on the radiograph is controlled within well defined limits. With computed radiography (CR), where the conventional screen-film combination is replaced by a storage phosphor screen, the process by which the digital image is presented on film restricts the resultant optical densities to an extremely tight range, independent of the air kerma incident upon the CR phosphor plate. The implication of this is that the measurement of optical density can not be used as a direct measure of AEC function, and a test such as Beam Quality Dependence cannot be performed in the conventional manner. To overcome this, the relationship between incident air kerma and a computer generated index supplied with the CR system was investigated for two commercially available systems, one supplied by Fuji Medical Systems and the other by Kodak. The relationship between the index and incident air kerma could then be used as a substitute for optical density in monitoring AEC performance with respect to beam quality. It has been determined that an inverse linear relationship exists between the incident air kerma and the Sensitivity Number for the Fuji CR system and that a logarithmic relationship exists between the Exposure Index and incident air kerma for the Kodak CR system. In order to quantify these relationships further, the dependence of Sensitivity Number and Exposure Index on beam quality and incident kVp have been investigated. Using the results from the above investigations, acceptable limits for both Sensitivity Number and Exposure Index, can be established to test for correct AEC function, with respect to Beam Quality Dependence. PMID- 9409021 TI - Mammography accreditation. PMID- 9409022 TI - Lymphangioma: imaging diagnosis. AB - Lymphangiomas are congenital malformations of the lymphatics that are curable by extirpation. Accurate delineation of lesion extension is important for pre operative diagnosis, surgical planning, and assessing recurrence. The radiologic findings were retrospectively evaluated to determine the imaging appearance of these benign tumours. The plain radiographs, barium meal, ultrasound, CT, and MR images of 18 patients with one or more pathologically proved lymphangiomas were reviewed. Plain radiography and barium study showed masses displacing adjacent organs. Ultrasound examination showed uni- or multilocular cystic masses with smooth, thin or irregular, thick walls. Enhancement of the cyst wall was variable on CT and MR studies. The CT density of the fluid ranged from -4 to 34 HU depending on the lipid content and the presence of haemorrhage. The cysts were isointense to muscle on T1-weighted and hyperintense to fat on T2-weighted MR images. The MR imaging delineated the tumour lesion extension more clearly than ultrasound and CT scans. Ultrasound, CT, and MR imaging are valuable for evaluating lymphangiomas. Magnetic resonance imaging allows accurate determination of lesion extension. PMID- 9409023 TI - Radiation dose in computed tomography of the pelvis: comparison of helical and axial scanning. AB - An anthropomorphic Rando phantom was used to compare radiation doses sustained during helical and conventional axial CT of the pelvis. The values obtained with the Rando phantom were validated against cadaveric phantoms, and show good agreement. For the authors' particular CT unit, helical scanning was found to deliver a lower radiation dose than conventional axial scanning. This was most prominent at 1.0-s tube rotation times (average dose ratio 1.24). For realistic scanning parameters and exposure factors, the ratio of radiation dose to pelvic organs can be expected to lie in the range of 40-100 mGy. The whole-body effective dose (ED) depends on selection of scanning parameters and patients anatomy. In a favourable case scenario, the ED for CT scanning of the pelvis in a male can be expected to be between 10 and 20 mSv if the scrotum is not included in the radiation field, while the ED in a female will be approximately 20 mSv. An examination of scatter radiation fall-off curves from a single slice shows that the spread of scatter radiation is only marginally affected by slice thickness. A total of 10-12 cm of human soft tissue acts as a good barrier against internal scattered radiation. The use of such scatter fall-off curves, together with manufacturers' dosimetry specifications, allows a fast estimate of absorbed dose. PMID- 9409024 TI - The 'spinnbarkeit phenomenon' of bile. PMID- 9409025 TI - Extrarenal angiomyolipomas of the perinephric space in tuberose sclerosis. AB - A case of bilateral angiomyolipomas of the perinephric space in a young Arab woman with tuberose sclerosis is reported. Multiple small angiomyolipomata were present in both kidneys and there was pulmonary involvement. The literature on this very rare occurrence is briefly reviewed. PMID- 9409026 TI - Percutaneous gastrostomy in radiologic practice. AB - Long-term gastrostomy tubes have a well-accepted role in providing nutritional support. Traditionally they have been placed by surgeons and by endoscopists. In the last decade, radiologists have come to play a major role in the placement of gastrostomy and gastrojejunostomy devices, and can usually do so as effectively and at lesser expense. A technique for placement is outlined, with a discussion of patient selection and complications. A review of the literature is provided. PMID- 9409027 TI - Cervical spine degenerative diseases: an evaluation of clinical and imaging features in surgical decisions. AB - In clinically severe cervical spondylosis, imaging plays a vital role in surgical decisions. A prime factor is acquired canal stenosis with cord compression. To validate this concept, the clinical and imaging features of 20 patients with spondylotic myelopathy and 24 with radiculopathy were retrospectively reviewed. All had computed tomographic myelography (CTM) as part of their clinical work-up. The patients' clinical severity was graded as mild, moderate and severe; the age, length of illness and a history of eventual surgery or otherwise were recorded. At the level of maximum compression the following parameters were obtained from the axial CTM images: surface area and ratio of the anteroposterior to the transverse diameter of the cord; subarachnoid space and vertebral canal areas. Data were statistically analysed. A significant association exists between surgery and increasing severity of symptoms (P = 0.04), and advancing age (P = 0.01). These associations hold true for myelopathy and radiculopathy. A strong association is present between surgery and the surface area of the cord (P = 0.01), being applicable to myelopathy only. The other parameters show no association with surgical decisions. It is concluded that with myelopathy a narrow cord area at the level of maximum compression, and moderate--severe functional impairment are indicators for surgical intervention. PMID- 9409028 TI - Endoluminal brachytherapy for recurrent laryngeal carcinoma. AB - Early-stage squamous cell carcinoma of the larynx is usually treated with local field radiotherapy. Surgery is used for salvage following recurrence. Further recurrences present a more difficult therapeutic problem which requires individualized management. The aims of local control, survival, maintenance of function and minimizing side effects all need to be balanced according to the site and extent of disease. The present case study looks at the management of a 54-year-old man with multiple recurrences from a squamous cell carcinoma of the larynx. It describes a technique of endoluminal brachytherapy using an iridium 192 wire spiralled around the outer part of a tracheostomy tube that achieves good local control while enabling self-insertion and self-cleaning during the procedure. The dose given was 2500 cGy at 5 mm over 25.2 h and was achieved with minimal early or delayed side effects. The patient had no further symptoms relating to the stomal recurrence until his death from metastatic disease 6 months later. PMID- 9409029 TI - Splenic irradiation for hairy cell leukaemia. AB - Splenic irradiation in the management of hairy cell leukaemia is previously unreported. A case is presented here to illustrate that splenic irradiation may be a useful addition to systemic therapies. PMID- 9409030 TI - Aggressive skin cancers in a cardiac transplant recipient. AB - Skin cancers following cardiac transplantation are a serious problem that may lead to significant morbidity and mortality. While the majority of skin cancers represent minimally invasive lesions that are treated adequately by local means, a small subset exhibits a potentially life-threatening natural history. The aggressiveness of these skin cancers, despite standard treatment, is indicative of the aggressive nature displayed by a subset of secondary malignancies following cardiac transplantation. The need for lifelong maintenance immunosuppression after transplantation is a major factor in the development of these secondary malignancies. Presented here is the case study of a cardiac transplant recipient who developed multiple aggressive squamous and non-squamous cell scalp skin cancers. PMID- 9409031 TI - Computed tomography features of lung parenchymal changes in pulmonary tuberculosis. AB - Twenty-five patients with active pulmonary tuberculosis were prospectively studied with CT. Three major parenchymal patterns were noted. The nodular opacities pattern was seen in all cases. Confluent consolidation was seen in 37% of patients and consolidation with associated loss of volume (CWALV) was seen in 69% of patients. High-resolution CT (HRCT) features of bronchogenic spread included (i) a centrilobular nodule or a branching linear structure (92.3%); (ii) bronchial wall thickening (61.5%); (iii) a 'tree in bud' appearance (92.3%); and (iv) poorly marginated 5-8 mm nodules (61.5%). Most of the patterns showed satellite lesions in the form of small nodules or peripheral areas of increased attenuation. Cavitation was most common in CWALV lesions. Bronchiectasis was a common accompaniment (81.3%), and its occurrence paralleled the distribution of parenchymal lesions. Associated pleural thickening was noted in half the cases. To conclude, distinctive parenchymal changes were noted with CT in cases of pulmonary tuberculosis, which may suggest the diagnosis in the appropriate clinical setting. PMID- 9409032 TI - Atypical computed tomography appearance of a duodenal diverticulum. AB - The typical CT appearance of a duodenal diverticulum is of a rounded air collection with or without fluid or contrast. An unusual case in which gas bubbles mixed with particulate matter mimicked faecal material in the large bowel is presented here. A similar appearance has been described in abnormally dilated small bowel, usually associated with obstruction, and is probably due to stasis of undigested food. PMID- 9409033 TI - Phaeochromocytoma with central nervous system manifestations. AB - A 35-year-old Samoan male presented with intermittent headaches and hypertensive episodes for several months. A subsequent left adrenal gland phaeochromocytoma was discovered and surgically excised. An MRI of his brain demonstrated periventricular, basal ganglia, and centrum semi-ovale infarction. We suggest that catecholamine excess and neuropeptide Y may contribute to intracerebral haemorrhage and infarcts associated with phaeochromocytomas. Additionally, our surgical approach in removing the phaeochromocytoma is discussed. PMID- 9409034 TI - An unusual consequence of stereotactic colloid cyst aspiration: case report. AB - The removal of a colloid cyst of the third ventricle by stereotactic cyst aspiration is frequently performed as an alternative to a transcortical transventricular or transcallosal operative route. A consequence of a CT-guided stereotactic aspiration, where residual colloid cyst material has been dislodged into the lateral ventricle and likened to an intraventricular mouse, is described. PMID- 9409035 TI - Frontal sinus carcinoma: a case report and review of the literature. AB - A rare case of frontal sinus carcinoma is reported. The patient developed early recurrence following surgery, and was managed by accelerated hyperfractionated radiation therapy and concurrent cisplatinum-based systemic chemotherapy. The patient has remained disease-free for 30 months following the end of treatment. PMID- 9409036 TI - Diffuse fatty infiltration of the liver: pitfalls in computed tomography diagnosis. AB - The presence of a fatty liver often complicates the interpretation of abdominal computed tomography (CT). Abnormalities in or adjacent to the liver, including dilated bile ducts, liver masses and subphrenic collections, may be masked by the fatty liver. Furthermore, normal structures may simulate pathological conditions. Five cases are presented to illustrate some of these diagnostic pitfalls. PMID- 9409037 TI - Review of the first 50 cases completed by the RACR mammography QA programme: phantom image quality, processor control and dose considerations. AB - The Mammography Quality Assurance Programme, recently established by the Royal Australasian College of Radiologists, has processed the first 50 applications. This programme, which closely follows the programme of the American College of Radiology (ACR), utilizes phantom film images, thermoluminescent dosimetry measurement of mean glandular dose, processor control charts, clinical images, equipment reports and required survey information to establish that a centre conforms to a minimum standard in mammography. The present paper describes the initial results of the first phantom images, dose measurements, processor control and survey information. A film review panel of six members has been trained in phantom film reading. Their evaluation of phantom films was compared with film readings by members of the ACR and was found to be in close agreement. Fifty films have been evaluated up to the present time with a failure rate of 26%. The major causes of failure were unacceptable film artefacts and poor contrast (as indicated by reduced fibre and mass visibility). A surprising result was the high failure in processing, where 23% of units reviewed had significant problems, including failure to keep the processor within required control limits. Only one centre recorded a mean glandular dose above 2 mGy with no centre over the 3 mGy limit. A review of the frequency of the quality control testing shows that the acceptance of quality assurance in mammography, while greater than in the initial stages of the ACR programme, is less than current US practice. These initial results for the accreditation process probably reflect an initial period of adjustment, as seen by the high pass rate achieved by centres that have resubmitted material to gain accreditation. PMID- 9409038 TI - Temporal lobe necrosis in nasopharyngeal carcinoma: pictorial essay. AB - Nasopharyngeal carcinoma (NPC) shows a high frequency of skull base erosion and intracranial spread. This tumour is usually treated with radiation therapy. The medial and inferior portions of both temporal lobes are included within the radiation portals. These areas are therefore potential sites of radiation-induced necrosis. It is important to recognize this complication and separate it from intracranial tumour recurrence because the treatments of these entities are different. PMID- 9409039 TI - 1996 Australian radiology workforce report. AB - A questionnaire was sent to all radiologists in Australia using a mailing list supplied by individual State Workforce subcommittee members. A reply rate of 72% was obtained. There are currently 1061 radiologists (1010.5 full-time equivalents) or 55.5 radiologists per million population, placing Australia in the mid-range compared with other OECD countries. This has increased slightly from 54.1 in 1994. There is a small but definite State variation. Utilizing current trainee numbers and traditional attrition rates, there is no projected change in these figures (55.3 in 2001), but the continued introduction of 'migrant' radiologists is postulated to cause an increase (56.6 with 25 migrants and 58.4 with 50 migrants in 2001). Analysis of work-practice indicates a performance rate of 14,100 procedures per year per practising radiologist. There is again a State variation. It is estimated that the total number of medical imaging procedures per 1000 population per year (rendered by radiologists) is 815. These latter two figures place Australia in the mid-range compared with the United States and Great Britain. General radiography, mammography, ultrasound, and CT are the most common procedures (in that order), and are performed by the largest proportion of radiologists. PMID- 9409040 TI - Radiotherapy for Hodgkin's disease in pregnancy. PMID- 9409041 TI - Intraventricular neurocytoma. PMID- 9409042 TI - Radiation oncology registrar training in Adelaide. PMID- 9409043 TI - Public perception of cross-infection control in dentistry. AB - Since the advent of HIV/AIDS at the beginning of the 1980s, concern has generated considerable impetus for change in cross-infection control procedures in dentistry. This process has been hastened partly by media coverage which, in tending to favour sensation over rational discourse, has played a not inconsiderable role in shaping public understanding and expectations. This study aimed to investigate public perceptions of cross-infection control in dentistry in Australia using a postal follow-up to the 1995 National Dental Telephone Interview Survey. The postal survey response rate was 85.2 per cent. Concerns about the procedures used by their dentist to sterilize instruments were reported by 13.3 per cent of respondents overall, and this was greater among non-health card-holders, individuals who mainly spoke a language other than English in the home, and those who reported a non-routine dental visiting pattern. Avoidance or delaying of dental visits due to the perceived cross-infection risk was reported by an overall 3.6 per cent of people, and this was higher among females and those who expressed concern about cross-infection control. The profession has a responsibility to ensure that information on the measures which have been taken to reduce the risk of cross-infection in dentistry is disseminated as widely and as clearly as possible so that undue public concern and avoidance of dental care are minimized. PMID- 9409044 TI - Lower lip paraesthesia following restoration of a second premolar tooth. Case report. AB - A forty year old female patient developed paraesthesia of the right side of her lower lip following the placement of an extensive pin-retained amalgam restoration in her lower right second premolar tooth. Radiographs indicated that the mental foramen was close to the apex of this tooth and it was assumed that postoperative pulpitis and periapical inflammation had caused the paraesthesia through the effects of pressure on the mental nerve. The paraesthesia resolved following endodontic treatment of the lower second premolar tooth and the patient has had no further signs or symptoms. PMID- 9409045 TI - The current status of low level laser therapy in dentistry. Part 2. Hard tissue applications. AB - While most applications of low level laser therapy (LLLT) in dentistry are directed toward soft tissues, in recent years there has been increasing interest in tooth-related or hard tissue applications of LLLT. This report provides an overview of applications of LLLT in the treatment of dentine hypersensitivity and pain arising from the periodontal ligament, and describes the phenomenon of lethal laser photosensitization and its applications in the treatment of dental caries. Technical aspects of LLLT equipment and safety concerns are also discussed. PMID- 9409046 TI - Trans-antral temporalis transfer for the repair of adult cleft palates. AB - Temporalis muscle transfer is a versatile technique frequently used for reconstructive procedures in the maxillofacial region. However the thickness of the pedicle may interfere with masticatory function when used anteriorly in the oral cavity. To repair full-length mid-palatal defects in fully dentate patients the flap can be passed through the maxillary sinus and combined with local repair of the soft palate, thus avoiding any occlusal trauma from the posterior teeth. The operation is a single stage procedure with low morbidity and few complications, and is a useful technique for repairing the large untreated clefts frequently encountered in developing countries. The procedure is used by members of the Australian and New Zealand Association of Oral and Maxillofacial Surgeons Bangladesh Project who have operated in Dhaka teaching hospitals on a regular basis since 1991. PMID- 9409047 TI - Adenomatoid odontogenic tumour (adenoameloblastoma). Case report and review of the literature. AB - An adenomatoid tumour was found in the anterior maxillary region of a 15 year old female patient. Two impacted teeth were found in the tumour. The lateral incisor found in the tumour was dilacerated, and the roots of the first premolar were resorbed. A review of the English literature indicated that 294 similar cases have been reported. PMID- 9409048 TI - Cysticercosis in labial tissue. Case report. AB - Cysticercosis is a condition in which a human acts as the intermediate host of Taenia solium, a pork tapeworm. The larvae infestation sites frequently include cerebral tissue, ocular organs, and muscles. The present case report describes a rare incidence of cysticercosis in the oral region. PMID- 9409049 TI - Enamel demineralization during orthodontic treatment. Aetiology and prevention. AB - The aetiology of enamel demineralization during fixed orthodontic treatment and its sequelae are discussed. A summary is given of the various methods available to assess the risk of demineralization prior to active treatment. The best preventive strategy would appear to be an assessment of risk factors prior to banding, coupled with fluoride rinses, regular reinforcement of oral hygiene, and dietary advice throughout the course of treatment. PMID- 9409050 TI - The analgesic efficacy of ibuprofen in periodontal surgery: A multicentre study. AB - The efficacy of a non-steroidal anti-inflammatory agent, ibuprofen, was evaluated in pain control following periodontal surgery. This type of agent acts peripherally by inhibiting the release of prostaglandins and minimizing the local inflammatory response. Thus there may be an advantage in pre-treatment administration of the drug so as to delay or even prevent postoperative pain. The study was multicentre, involving a Public Hospital Periodontal Unit, two specialist periodontal practices in Sydney, NSW, and two in Canberra, ACT. One hundred and twenty-seven patients who were to undergo periodontal surgery were randomly given either two 200 mg tablets of ibuprofen or two matching placebo tablets at least 30 minutes before administration of local anaesthesia. The procedure was double blind: neither the patient nor the clinician was aware of the tablet identity. Postoperatively, all patients were given labelled ibuprofen for pain relief, but were randomly divided into two groups: As directed who were instructed to take the drug regularly for two days postoperatively, and As required, who were to take the drug only if needed for pain relief. All patients completed a diary recording quantity and time of medication, and regular assessment of pain experience utilizing a visual analogue scale. The As directed group showed no significant difference in pain experience between pre-operative and post-operative only medication, but the As required group experienced significantly less pain and requirement for medication if the ibuprofen was administered pre-operatively. PMID- 9409051 TI - Tissue radiation dosages using the RVG-S with and without niobium filtration. AB - Tissue doses for a modified Rando head- and-neck phantom were measured for imaging with speed group E film with standardized aluminium filtration and the RVG-S both with and without added niobium filtration. Cylindrical holes drilled into the phantom's tissue-equivalent material permitted the placement of a small ionization chamber into anatomically correct sites representing the thyroid, parotid, submandibular and sublingual glands. To establish the necessary cone positions, angulations and time settings for each exposure, diagnostically acceptable images of six teeth, representative of different intraoral regions, were made for a DXXTR mannequin. Entrance and exit points were marked and transferred to the phantom to allow reproducible repeat exposures. The RVG-S provided reductions in average skin entrance dose of 31 per cent to 39 per cent with standard aluminium filtration and 51 per cent to 60 per cent with the addition of niobium filtration to attenuate the beam. While dose reductions relative to E-speed film usage were found for deep tissue sites, these were site and projection specific. The cumulative reduction from use of the RVG-S without niobium filtration was 32 per cent. It was 42 per cent with additional niobium filtration. It should be noted, however, that adding niobium filtration resulted in increased dosages to the deeper soft tissues such as the thyroid gland. PMID- 9409052 TI - Growth inhibition of oral bacteria related to denture stomatitis by anti-candidal chalcones. AB - In the antimicrobial therapy of denture stomatitis, it is desirable to inhibit the growth of not only the primary causative organism, Candida albicans, but also other oral bacteria closely associated with the condition. Three synthetic anti candidal chalcones were characterized and compared for their additional activity in inhibiting these causative bacteria. Among the tested chalcones, 2,4,2' trihydroxy-5'-methylchalcone showed the highest activity for different Gram positive bacteria. It inhibited the growth of streptococci, staphylococci and lactobacilli at 25.0-50.0 micrograms/mL which was lower than or comparable to its minimum inhibitory concentration for candida. It functioned with a bactericidal action and leaked 260 nm-absorbing substances from the streptococcal cells. The antimicrobial activity of 2,4,2'-trihydroxy-5'-methylchalcone against both primary and secondary causative agents suggests it could be useful as a potent therapeutic agent in denture stomatitis. PMID- 9409053 TI - Deregulation, competition policy--where do we go from here? PMID- 9409054 TI - Infectious waste storage. PMID- 9409055 TI - Elsdon Storey Memorial Lecture. Dentistry and the world of children. PMID- 9409056 TI - Case management study: methotrexate side effects--or not? PMID- 9409057 TI - Human immunodeficiency virus infection and rheumatic disease. PMID- 9409058 TI - Helicobacter pylori and nonsteroidal antiinflammatory drugs: partners in crime? PMID- 9409059 TI - Health promotion and early detection of cancer in older adults: needs assessment for program development. AB - The greatest risk factor for developing cancer is age, yet little is known about the cancer-related knowledge, attitudes and beliefs of the older adult (age > 55 years). A community-based needs assessment was conducted to understand these dimensions in a large metropolitan community. Ten focus groups (n = 158 older adults) and 9 individuals were interviewed. Content analysis for the audiotaped sessions was completed. The study participants focused on "being active" and living a healthy lifestyle. Many worried about illness interfering with their ability to do what they wanted to do. Many had had exposure to cancer through family members or friends, but still had many unanswered questions about cancer. Age was not seen as a risk factor for cancer, and a range of attitudes existed regarding cancer prevention and early detection. Overall, despite fearing cancer, participants thought older adults needed to know about cancer and suggested a wide range of approaches to disseminate information effectively to older adults. PMID- 9409060 TI - Decision making by elderly patients with cancer and their caregivers. AB - This study explored the scope of decisions encountered by elderly cancer patients and/or their family caregivers, and the types of decision-making assistance requested and required within one practice setting. Semistructured interviews were conducted with five cancer center nurse coordinators (CCNCs). The CCNCs were interviewed weekly for 16 weeks to identify decision-making topics addressed, assistance requested, and perceptions of assistance required during telephone conversations. The CCNCs' reports of 41 telephone conversations revealed 44 specific decision-making topics. Content analysis uncovered 11 categories: symptom management, use of chemotherapy, ancillary choices selection of a medical provider, planning for end-of-life care, alternative therapy, vacation planning, weekend-pass planning, discharge planning, family survivor issues, and involvement of adult children in the elder's care. Elderly patients and/or their family caregivers requested information and assistance with making decisions. CCNCs perceived that callers also needed information clarification, reassurance about their decisions, a listener, permission to change the treatment regimen, and help with communication among health professionals, the elderly patient, and the family. PMID- 9409061 TI - Quality of life in breast cancer. Part I: Physical and social well-being. AB - Almost 2 million breast cancer survivors reside in the United States. An increase in consumer advocacy and media attention to this disease has helped bring breast cancer survivorship to the forefront of public attention. This has led to increased attention on quality of life (QOL) issues for these survivors of breast cancer. This two-part article presents the results of a qualitative, descriptive study evaluating the QOL of 21 breast cancer survivors. This study is based on our conceptual model of QOL including physical, psychological, social, and spiritual well-being. Part I of this article describes the impact of breast cancer on the physical and social domains of QOL based on in-depth interviews with breast cancer survivors. PMID- 9409062 TI - Supportive therapies for cancer chemotherapy patients and the role of the oncology nurse. AB - Cancer chemotherapy often causes severe side effects, such as neutropenia, nausea and vomiting, and oral complications, which adversely affect patients' quality of life and may interfere with treatment success. A number of supportive therapies, such as colony stimulating-growth factors and antiemetics, have been developed to ameliorate these side effects, however, but often are underutilized. Oncology nurses, who serve as liaisons between oncologists and patients, can have a positive effect on patients' quality of life by educating them about potential side effects and the availability of supportive therapies, and by bringing patients' quality of life concerns and priorities to the attention of physicians. This article reviews the side effects of chemotherapy and the supportive therapies currently available to treat them and explores the role of oncology nurses as advocates for improved quality of life for chemotherapy patients. PMID- 9409063 TI - A descriptive study of stress management in a group of pediatric oncology nurses. AB - Pediatric oncology nursing is associated with highly stressful and emotional situations. This article describes and discusses major sources of occupational stress among a group of nurses participating in a stress management group. The stress sources for these nurses were preoccupation with death and dying, the professional image of the oncology nurse, the nurse as fighter in the war against disease and death, the nurses' perceived isolation from the medical staff, the nurses' perceived inferior professional status compared with that of physicians, emotional overinvolvement with patients and families, suppression of anger, and difficulties in balancing work and home demands. The following factors are suggested as major contributors to the nurses' stress and burnout: increased tendency for irrational-dysfunctional thinking styles (mainly "demandingness" and "awfulizing"), diffuse boundaries between nurses and patients, low professional self-efficacy, and wide prevalence of military metaphors. PMID- 9409064 TI - Feelings of powerlessness in relation to pain: ascribed causes and reported strategies. A qualitative study among Dutch community nurses caring for cancer patients with pain. AB - This qualitative study focused on the causes for the feelings of powerlessness experienced by dutch community nurses in caring for cancer patients with pain. In addition, the study focused on the strategies community nurses employed to cope with feelings of powerlessness. Semistructured interviews revealed the following ascribed causes for feelings of powerlessness: problems in communication between community nurses and patients and between community nurses and personal physicians, dilemmas concerning physical care and opiates, and discrepancies between community nurses' pain management goals and what can realistically be achieved. The main strategies for reducing feelings of powerlessness were sharing feelings with the patient and colleagues, increasing knowledge and skills, and standing back from the situation. PMID- 9409065 TI - Side effects of CHOP in the treatment of non-hodgkin's lymphoma. AB - Cyclophosphamide, doxorubicin, vincristine (Oncovin), and prednisolone (CHOP) has for many years been the standard chemotherapeutic regimen for patients with aggressive non-Hodgkin's lymphoma. Published data for side effects experienced by patients undergoing CHOP chemotherapy in the treatment of non-Hodgkin's lymphoma are limited and inconsistent. No broad descriptive work appears to have been carried out. This study aimed to describe the range of problems experienced by patients receiving CHOP and to estimate incidence and severity of side effects over the treatment period. Data were collected at each treatment cycle via a 75 item self-report questionnaire, with severity of each side effect graded on a 5 point scale. The instrument has previously been shown to be reliable and valid. Nineteen participants received 99 cycles of CHOP and returned 74 questionnaires (response rate = 75%). Patients reported a total of 80 side effects. Alopecia was the most common problem, with all patients experiencing some hair loss by cycle 3. Fatigue was the second most common side effect (incidence = 77%) and taste change the third (incidence = 74%). Patients judged postchemotherapy nausea to be the "most troublesome" problem, followed by fatigue, taste change, constipation, and difficulty sleeping. Both nausea and fatigue were most problematic in the first part of the treatment course. These results indicate that patients receiving CHOP experience a wide range of problems, many of which merit further investigation. PMID- 9409066 TI - Infection control in child care settings. AB - Over one-third of all under five year old Australian children use some form of licensed child care. The majority of research on infectious diseases in children using care, mainly emanating from North American and Scandinavia, suggests that children in preschool or long day care suffer more frequent infections and more days of illness than those cared for at home or in family day care. In order to minimise these risks it is necessary to apply infection control principles. In this article infection risk factors are outlined and recommendations for immunisation, preventative practices, the use of antibiotics and outbreak management are presented. PMID- 9409067 TI - Communicable diseases surveillance. PMID- 9409068 TI - Cultural babel: the challenge of immigrants to the helping professions. AB - Immigrants to the United States often experience sociocultural stress not easily mitigated by the culturally individualistic and Western endopsychic probing methods of therapy. The vast majority of immigrants hold communal attitudes and holistic beliefs, whereas American therapists use treatment methods that endorse self-sufficiency and individualistic values. Parsimonious, psychoeducational, and less ego-threatening strategies are proposed--supportive, directive, didactic, and reality-oriented approaches that are consistent with the treatment perspective of immigrant populations that commonly hold collective rather than individualistic views. The proposed goal of intervention is to help immigrants become bicultural citizens, by enabling them to adopt American coping skills without surrendering those of their own culture. PMID- 9409069 TI - Immigration patterns, social support, and adaptation among Korean immigrant women and Korean American women. AB - There are little empirical data available on the mental health and social functioning of Korean American Women (both native U.S. born and foreign Korean born U.S. residents, inclusive). State-of-the-art research used to inform social work practice is exploratory descriptive. With the goal of contributing to the social work knowledge base regarding this understudied population, this article uses an emic understanding and approach to examine immigration patterns, social support networks, and issues around adaptation experienced by Korean American women. Issues examined include gender role disruption, limited use of social services, and evidence of depressive symptoms in Korean American women and subsequent risk of substance abuse, suicide, battering, loss of employment, deficits in parenting, and mental health problems. Focus on these areas of functioning suggests the need for development of culturally competent community, family, individual, and organizational-level intervention strategies. PMID- 9409070 TI - The relationship of DSM diagnostic criteria and Gough's Prejudice Scale: exploring the clinical manifestations of the prejudiced personality. AB - The relationship of psychopathology, symptoms of personality disorder, and outgroup prejudice was examined with 193 outpatient psychotherapy clients. Primary DSM-IV diagnosis, General Adaptive Functioning (GAF) scores, personality disorder criteria, and Minnesota Multiphasic Personality Inventory (MMPI) scale scores were examined in relationship to Pr (Prejudice) Scale scores and client outgroup attributions. Results of a 3 x 10 Multivariate analysis of variance (MANOVA) indicated that clinician ratings of outgroup bias were significantly related with the Axis II criteria for Paranoid, Borderline, and Antisocial disorders. MANOVA results for ratings of outgroup bias and MMPI scores did not yield a significant multivariate effect; however, significant univariate ANOVA results were found with the MMPI F, HS, PD, and MA scales. Computed univariate ANOVA results indicated that Pr Scale scores did not significantly vary between primary Axis I and Axis II DSM-IV diagnosis, but did yield a significant difference for (categorical) diagnosis by Axis II Cluster groups. Both Pr Scale scores and clinician ratings of client outgroup bias were significantly related to greater psychopathology, as reflected by lower GAF scores assigned at the initiation of treatment. Findings provide preliminary evidence of the relationship of traits of personality disorder, as characterized by impulsivity, relational disturbance, and affective lability, to outgroup prejudice with a clinical population. PMID- 9409071 TI - The phenomenology of ritualized and repeated behaviors in nonclinical populations in the United States. AB - The question of whether ritualized and repeated behaviors might be part of a person's "normal" repertoire, providing them with feelings of self-efficacy, is explored in a phenomenological study in which nonclinical subjects who engage in ritualized and repeated behaviors describe their behaviors, and reflect on their feelings before, during, and after engaging in these rituals. Results seem to support the hypothesis that engaging in these behaviors is perceived as reducing feelings of anxiety, fear, and discomfort, and increasing feelings of control and security. Subjects also report that they would feel anxiety and lack of control if confronted with the possibility of interference with rituals. Results contribute to the conceptualization of ritualized and repeated behaviors as coping strategies in an individualized society, which function to alleviate anxiety and promote self-efficacy. It is suggested that this paradigm be used to explore obsessive-compulsive disorder in clinically diagnosed populations of ritualizers, in terms of etiology and for purposes of treatment. PMID- 9409072 TI - Psychiatric symptoms in offspring of within vs. across racial/ethnic marriages. AB - A large number of adolescents of interracial ancestry (parents comprising various combinations of African-American, American Indian/Alaska Native, European American, Chinese, Filipino, Hispanic, Japanese, Korean, Puerto Rican, Samoan, and Tongan ancestry) were contrasted with a monoracial European-American sample in the degree to which they reported symptoms of depression, anxiety, conduct disorder/aggression, and substance abuse. The adolescents of interracial ancestry were subdivided into three groups in terms of parental ancestry: both parents of interracial ancestry, one parent of interracial and the other of monoracial ancestry, and both parents of monoracial but different ancestries. The interracial ancestry groups did not differ significantly from one another or from the European American sample in terms of symptom scores. PMID- 9409073 TI - Clinical presentations of depression in African American and white outpatients. AB - The objective of this study was to compare the nature and severity of depressive symptoms in moderately depressed, medically healthy African American and White patients. Twenty age- and gender-matched subjects from each ethnic group who met criteria for a major depression were assessed with structured interviews, and their depression was evaluated with the Hamilton Depression Rating Scale (HAM-D). Overall severity of the depression was comparable between groups. When the individual HAM-D items were grouped into factors, Whites showed significantly more articulated/observed mood and anxiety symptoms, whereas African Americans had significantly more diurnal variation to their depression. There were no differences on other neurovegetative symptoms. These results are discussed in the context of past studies, which often used very heterogeneous populations not matched for socioeconomic status, and included those with comorbid psychiatric and medical illnesses. Although our sample size was relatively modest, the results suggest that clinicians should be aware of potential differences in symptom presentation when treating patients from different ethnic groups. PMID- 9409074 TI - Volume and topographical changes of the basolateral complex during the development of the rat's amygdaloid body. AB - Volume and topography of basolateral complex during development and maturation in fetal and postnatal rat brains were studied using morphometrical methods. 39 rat brains of various ages were fixed in formalin, frozen and cut into 25-micron thick sections and stained with cresyl violet. In cresyl violet preparation the basolateral complex appeared first on 17th prenatal day and it was composed of two homogenous parts--lateral and basolateral nuclei. On about 7th postnatal day each of these nuclei divided into two parts--the first one into the dorsolateral and ventromedial part, while the second one--into the anterior and posterior parts. Morphometric investigations showed that volume of the basolateral complex and its parts underwent the biggest changes up to 14th postnatal day, and we suspected that during this time it was more sensitive to pathological changes. After that day the volume of the basolateral complex had changed only a little. PMID- 9409075 TI - Cytoplasmic dense bodies in pig pinealocyte during postnatal development. Quantitative, ultrastructural study. AB - Cytoplasmic dense bodies, conspicuous structures of pinealocytes in the pig pineal gland were ultrastructurally examined during postnatal development from the first day after birth to puberty. The morphometric analysis was employed to estimate the quantitative changes. Process of progressive increase in the relative volume with simulatenous differentiation of structure of the bodies was observed. PMID- 9409076 TI - Fine structure of the capillaries in the spinal cord of human embryos at 7th week. AB - Study were made in three human embryos at developmental stages 18 and 19 (7th week). It was shown that the wall of the capillaries is formed by one or two endothelial cells, which are separated by perivascular space from the glial cells. Endothelial cells possessed cilia projecting into the vascular lumen and are connected through the zonulae occludentes. PMID- 9409077 TI - A microscopic view of false tendons in the left ventricle of the human heart. AB - Research was conducted on material consisting of 45 fetal, newborn, infant and adult human hearts. False tendons in fetal, newborn and infant hearts were made up of mainly heart muscle tissue. False tendons of the tendon attachment to the interventricular septum. These false composed of muscle tissue and connective tissue in various proportions. Most of the connective tissue was observed in this age group in the area of the tendin attachment to the interventricular septum. These false tendons turned out to be very richly vascularized. In some of them, elements of the conductive system was confirmed, being an extension of the left branch of the bundle of His. This may confirm the role of false tendons in heart arrhythmias. PMID- 9409078 TI - Vascularization of the brain in guinea pig III. Vascular architecture of the medula oblongata, pons, and cerebellum. AB - The blood vessels of the medulla oblongata and pons are arranged into anterior (medial), lateral, and posterior groups. Particular part of the brain receive also blood from several sides (medial, lateral, posterior) what secures rich capillary net and vascular loops. Venous blood is drained by veins running in the same directions. The richest capillary nets are found within brain stem nuclei. The cerebellum is supplied by three cerebellar arteries. PMID- 9409079 TI - Vascularization of the brain in guinea pig. IV. Angioarchitectonics of the tectum, tegmentum, and crura mescencephali. AB - Out of three parts of the midbrain the richest vascularization is observed in the tegmentum in which are localized many nuclei. Particularly abundant blood supply shows the main nucleus of the oculomotor nerve. Less vascularized is the central oculomotor nucleus, and scanty blood vessels are found within the autonomic nucleus of the oculomotor nerve. Dense vascular net is found in the supratrochlear nucleus which lies in the central gray substance of the midbrain. Also, dense vascular net is noted within the red nucleus, trochlear nucleus, and the dorsal raphe nucleus. Within the substantia nigra less vascularized is its compact part. Very weak vascularization is observed in the periaqueductal gray and in the crura cerebri. Within the tectum more blood vessels are found in the inferior colliculus as compared with the superior colliculus. Within the crura cerebri the blood vessels do not form vascular net and they ascend into tectum and tegmentum. PMID- 9409080 TI - Vascularization of the brain in guinea pig. V. Angioarchitectonics of the thalamus, telencephalon and internal capsule. AB - The main blood supply of the thalamus constitute internal branches of the posterior cerebral, posterior communicating, middle, and choroid arteries. Branches of these arteries from three groups of vessels: medial, lateral, and posterior. Considering the number of vessels, their diameter and direct origin from the cerebral arterial circle, the thalamus is best vascularized structure of the brain. The telencephalon is supplied by rami of the anterior and middle cerebral, choroid, posterior communicating, and posterior cerebral arteries. The internal capsule and telencephalic nuclei are supplied by the internal branches of the same vessels as the telencephalon as well as by the cortical rami of the middle and posterior cerebral arteries. All these vessels from anteromedial, anterolateral, and superolateral groups. Between the internal and cortical branches anastomotic vessels are seen. PMID- 9409081 TI - The complexity of molecular genetic research in psychiatric disorders: advances and pitfalls. AB - This article describes the major techniques in molecular genetics, i.e., parametric and nonparametric linkage analyses, association studies and quantitative trait loci approaches. Some major molecular findings in schizophrenia, bipolar affective disorder and Alzheimer disease are presented. Aspects of the complexity of molecular genetic research in psychiatric disorders are discussed, including the relationship between genotype and phenotype, and between etiology and pathophysiology. Molecular findings in two genetic neuropsychiatric disorders, fragile X syndrome and Huntington's disease are described. These findings provoke some critical thoughts regarding future directions in psychiatric molecular research. PMID- 9409082 TI - Oral maternal inositol supplementation does not increase rat conceptus inositol levels. AB - Lithium (Li) is an effective drug for prophylaxis and treatment of major affective disorders. It is teratogenic both to animals and human beings. Depletion of inositol is associated both with lithium side effects and teratogenesis. There is no direct evidence showing that in humans Li teratogenesis is associated with the phosphoinositol (PI) cycle. It is conceivable that the teratogenic effect of Li in humans is also associated with inositol depletion and therefore is amenable to inositol supplementation. To test the hypothesis that oral inositol may cross the placental barrier and may be useful as a protective supplement to lithium therapy during pregnancy, we studied the effect of 2.5% inositol in drinking water on embryonic inositol levels in rats. There was no effect on fetal inositol concentration. However, weight of embryos of mothers receiving inositol was significantly higher. These data do not support the concept that inositol supplementation may be useful in preventing human Li-induced teratogenesis. PMID- 9409083 TI - Subjective experience and attitudes towards participation in clinical trials among patients with anxiety disorders. AB - Fifty-two subjects with panic disorder and generalized anxiety disorder were interviewed 12-18 months after they completed an acute treatment study (9-11 weeks) with alprazolam followed by blind and controlled drug discontinuation. Patients were questioned about the severity of their anxiety disorder and degree of functional impairment at the time of follow-up, as well as about their subjective experience as participants in a clinical trial. At the time of follow up 78% of the patients reported none or minimal anxiety symptoms and 89% had none or only minimal functional deficit, as compared to 100% reporting moderate to severe anxiety symptoms and 57% reporting significant functional deficit before entering the study. Overall, patients felt that the participation in the study was a very positive and beneficial experience and 64% said they would not hesitate to participate in a clinical trial in the future. PMID- 9409084 TI - Personality genetics. AB - Although 30-60% of the variance in many personality traits is inherited, until recently little was known about the responsible genes. Preliminary studies of family history in bipolar disorder and of X-linkage of personality traits in color-blindness suggested a "quantitative trait locus" (QTL) approach to the genetics of normal personality. In methodically similar but independent studies of 124 Israeli and 315 American normal volunteers we showed an association between the dopamine D4 receptor gene (D4DR) and the personality trait of novelty seeking. In the Israeli sample we also found an interaction between the D4DR gene and the serotonin 2C receptor gene (5-HT2C) with a marked effect on the trait of reward dependence. Further investigation of genes for personality traits may suggest links between normal personality and psychiatric illness. PMID- 9409085 TI - Serum neuroleptic levels during reduced dose fluphenazine decanoate maintenance therapy. AB - Forty-one remitted and chronically psychotic schizophrenic out-patients completed a two-year clinical trial during which they were assigned, on the basis of their clinically determined maintenance dosages, to one of two reduced, fixed-dose fluphenazine decanoate (FD) regimens: 35 mg/4 wks (19 patients) or 10 mg/4 wks (22 patients). Eighty-one percent of chronically psychotic patients, who represented 74% of the high dose group, relapsed, in comparison with only 38% of remitted patients (p < .001), who represented 86% of the low dose group. During this study serum neuroleptic levels were assessed, using the radioreceptor assay, before the administration of each FD injection and whenever a patient relapsed. Overall, 334 serum neuroleptic activity measurements were performed. Serum neuroleptic levels were detectable in all patients and were higher, although not significantly, in the 35 mg/4 wks group. The dichotomous clinical outcome of chronically psychotic and remitted patients occurred within the framework of essentially similar serum neuroleptic levels. These findings suggest that: 1) serum neuroleptic levels can be monitored during low dose FD treatment, 2) the poor maintenance therapy outcome of chronically psychotic patients cannot be accounted for by inadequate neuroleptic bioavailability, 3) a majority of remitted FD maintained patients retain their clinical response at serum neuroleptic levels lower than those initially attained at steady state. PMID- 9409086 TI - Taste- and odor-reactivity in heroin addicts. AB - Opiates in general, and heroin in particular, are known to induce compulsive drug seeking and drug-taking behavior. Addiction is accompanied by psychobiological processes which may distort perception of sensory stimuli. Gustatory and olfactory stimuli are hedonically polarized and therefore most appropriate for the assessment of the organism's reactivity to "useful" and "harmful" chemosensory events. Previous studies revealed that psychophysical self-estimates and reflectory facial expressions mirror with comparable reliability the hedonics of the perceived taste and odor sensations. In the present study both cognitive verbal and reflectory facial expressions of a group of: a) heroin addicts were recorded and compared to those of a group of b) detoxified former addicts and to c) a group of matching controls. Results show that all three groups differentiate between pleasant, indifferent and aversive tastes and odors. Active addicts estimated sweet taste and savory smells as being somewhat more pleasant, and bitter and sour tastes and a putrid odor as less unpleasant than did the other two groups. The reflectory facial displays of addicts were less expressive and discriminative than those of the two other groups. Taste- and odor-induced facial displays are known to be controlled primarily by the brainstem. The findings indicate that heroin-addiction affects brain-mechanisms, which mirror taste- and odor-hedonics. Modulation of the phylogenetically ancient, sensory-motor coordinations was found to be of a different pattern than that of the cortically controlled cognitive reactions. PMID- 9409087 TI - Selective effects of MAO inhibition on peripheral benzodiazepine receptor binding in the mouse. AB - Monoamine Oxidase (MAO) and the peripheral benzodiazepine binding site (PBR) share a close physical proximity to each other in the outer mitochondrial membrane. Furthermore, MAO activity and the density of PBR sites are affected by stress; benzodiazepines may influence stress-induced changes in MAO activity. In view of the close physical association between MAO and the PBR, we examined the effects of chronic administration of selective and nonselective MAO inhibitors to mice on the specific binding of 3H-Ro5-4864 and 3H-PK-11195 to crude membranes prepared from kidney, heart and liver. Chronic MAO inhibition was associated with alterations in PBR binding in all three tissues; however, in heart and liver changes were not detectable with 3H-PK-11195. Perhaps, the ability to discern changes with 3H-Ro5-4864 that are not detectable with 3H-PK-11195 reflects a functional change in the "activity" of the PBR site in heart and liver that is elicited by chronic MAO inhibition and mediated by a change in the "conformation" of the protein that is detected with 3H-Ro5-4864. Importantly, iproniazid, the nonselective MAO inhibitor, caused changes in PBR binding in all three of the tissues. PMID- 9409088 TI - Bipolar affective disorder associated with Klinefelter's syndrome--a case report. AB - Several cytogenetic surveys and literature reviews have suggested a possible association between Klinefelter's syndrome and bipolar illness. We describe a manic patient with Klinefelter's syndrome who was treated successfully with lithium. Our patient represents the 32nd reported case of bipolar disorder in an individual with a known chromosomal abnormality, the 15th with Klinefelter's syndrome, and the 11th with a karyotype of 47XXY. PMID- 9409089 TI - Possible pitfalls in the diagnosis of acute myocardial infarction in acutely psychotic patients. AB - The disturbed communication of the acutely psychotic patient with Emergency Room (ER) personnel can occasionally lead to a missed or delayed diagnosis of acute myocardial infarction. Conversely, miscommunication may also lead to a false positive diagnosis of acute myocardial ischemia or infarction. We describe a patient who failed to report lithium ingestion because of his psychotic and toxic state. Moreover, the patient was agitated and was treated with intramuscular haloperidol. These factors produced electrocardiographic changes due to the lithium and a rise in serum creatine kinase due to the intramuscular haloperidol, two criteria which could mislead an inexperienced observer to make the false diagnosis of acute myocardial ischemia or infarction. Lithium related electrocardiographic abnormalities and the causes for elevated creatine kinase isoenzymes are reviewed. PMID- 9409090 TI - Risperidone in the treatment of catatonia in a schizophrenic patient. PMID- 9409091 TI - Domestic social integration and suicide in Israel. PMID- 9409092 TI - Demonstrating the added value of community health nursing for clients with insulin-dependent diabetes. AB - The importance of demonstrating that nursing care adds value to client outcomes is discussed broadly as well as within the dimensions of a community health nursing outcomes study. The study examined the effect of a community health nursing intervention on the health behaviors and health status of adults with insulin-treated diabetes as measured by client outcomes using a two-group repeated measures experimental design. The findings indicate that the community health nursing intervention significantly enhanced the self-reported self-care behaviors of blood glucose testing, complication management, nutrition regimen adherence, and reporting foot change in the experimental group. No differences were found between the groups in outcomes for dietary adherence, foot care, blood glucose levels and overall diabetes knowledge, metabolic control, or functional health status. Challenges for community health nurse researchers engaging in client outcome studies are delineated and discussed. PMID- 9409093 TI - Using a nursing framework to establish a nurse-managed Senior Health Clinic. AB - This article describes the organizing framework and evaluation component of a newly initiated nurse-managed Senior Health Clinic that is being collaboratively run by a school of nursing and a senior service agency in Central Florida. Background is provided on choosing a partner agency; negotiations; developing a mission statement, goals, and operation plans; determining the target audience; and marketing strategies. The clinic targets residents who are 55 years old and older and who do not currently have a primary care provider. Cox's (1982) Interactional model for Client Health Behavior provides the framework for care delivery and all process and outcome evaluation activities. As such, the model provides an innovative nursing focus to the delivery of primary care services. Client characteristics are used to form the components of the client data collection tool, which was developed to provide baseline information on seniors who are using the clinic. Three elements of client professional interaction are included in the Health Care Form, which documents the nurse practitioner services that were provided: health information, affective support, and professional technical competencies. The Client Satisfaction Tool measures the client's satisfaction with each of the elements of client-professional interaction that are theorized to influence health outcomes. Information management is facilitated by a computer program that enables staff to enter the information directly into the computer. Data collection is ongoing and will quantify the type and extensiveness of services provided and the quality of care at the Senior Health Clinic. PMID- 9409095 TI - Acupuncture, fools, and horses. PMID- 9409094 TI - Preventing abuse to pregnant women: implementation of a "mentor mother" advocacy model. AB - Abuse to pregnant women is common and can result in complications to maternal and child health. Although screening and detection of abuse in primary health care settings is becoming more commonplace, intervention models that include community outreach have not been developed or tested. An advocacy model was developed and tested for pregnant abused women by melding research on advocacy programs for abused women exiting shelters with the principles of home visitation used to improve outcomes to pregnant women. Advocacy was offered by "mentor mothers," who were residents of the project's service area. The advocacy consisted of weekly social support, education, and assisted referrals to pregnant women identified as abused as part of routine screening offered at the first prenatal visit to a public health clinic. Effectiveness of the advocacy intervention was measured as contact success rate, number and type of advocacy contacts, and number and type of referrals made to the first 100 women to complete the advocacy program. The mentor mother advocates were successful in contacting the abused woman 33% of the time, regardless of whether a telephone call, home visitation, or in-person meeting was attempted. The average number of advocacy contacts was 9.2 (SD = 7.6) with the majority (74%) being via the telephone. The average number of referrals per woman was 8.6 (SD = 7.6) with the largest percentage (38%) being for medical services. Outreach advocacy as an intervention model for pregnant abused women is recommended. PMID- 9409096 TI - Is the cost of preservative-free opioids for intraspinal use justified? PMID- 9409097 TI - Symptom prevalence in the last week of life. AB - Palliative care is the management of patients with progressive, far-advanced disease for whom the prognosis is limited and the focus of care is quality of life. During the last days of life, it is important to redefine the goals, as previously present symptoms may increase and new symptoms may appear. To assess these symptoms, 176 patients were evaluated. A questionnaire evaluated symptoms during the last week of life and compared these prevalences with those at the first evaluation. The patients comprised 121 men and 55 women. The mean age was 67.7 years. Metastases were present in 66.5% and were multiple in 52%. The most frequent symptoms at the end of life (> 50%) were anorexia, asthenia, dry mouth, confusion, and constipation. The majority of patients died at home (64.2%). We observed good control of "reversible" symptoms, but many symptoms were difficult to control at the end of life. Symptom assessment is important in this population. PMID- 9409098 TI - A Canadian survey of issues in cancer pain management. AB - We report the analysis of a cancer management survey mailed to a representative group of health professionals in 1994. The goals of the study were to gather information on cancer pain treatment practices, and to obtain health professional views on obstacles to ideal pain management. The survey, designed by a working party of pharmacists, nurses and physicians, was distributed to 14,628 physicians. A total of 2,686 physicians responded to the survey, including 39% of medical or radiation oncologists, and 18.19% of physicians who listed their primary interest as Family Medicine. Reflecting the modest emphasis placed on palliative care and cancer pain management in the current Canadian milieu, 67% of physicians rated their past teaching experience as only "fair" or "poor." Lack of exposure to pain education was reflected in the response to a series of hypothetical case scenarios exploring physician choices in managing severe cancer pain. For example, in the initial management of a cancer patient with severe pain, 50% of physicians would not use a strong opioid in the absence of other contraindications to opioid use. A wide variety of analgesics and non pharmacologic techniques is available to Canadian physicians to assist patients with pain. Few physicians identified the unavailability of analgesics or analgesic techniques as limiting factors in pain management. We conclude that greater emphasis should be placed on pain education in our training programmes. We suggest that further surveys of this type, sponsored by our provincial colleges and medical organizations, can provide feedback which will enhance the adherence by Canadian physicians to published guidelines for pain management. PMID- 9409099 TI - Measurement of neonatal responses to painful stimuli: a research review. AB - The measurement of neonatal responses to painful stimuli remains a significant clinical problem. Although numerous measures have been evaluated, instruments that are valid, reliable, and clinically feasible are not yet available. The purpose of this paper is to critique the studies that have been done using biochemical, physiological, and behavioral measures to evaluate neonatal responses to painful stimuli. Specific issues regarding measurement in premature and critically ill neonates are emphasized. The intent of this review and critique of the literature is to stimulate additional research into the assessment of neonatal pain. PMID- 9409100 TI - "Just one look...". PMID- 9409101 TI - "A 43-year-old black male ...": conventional wisdom, racism, or noise? PMID- 9409102 TI - Polycarbonate is great, but... PMID- 9409103 TI - Eye signs and tic disorders: Gilles de la Tourette's syndrome. AB - BACKGROUND: Eye tics are among the most common manifested symptoms of tic disorders, including Gilles de la Tourette's syndrome (GTS). METHODS: A review of medical literature or ocular signs and symptoms of tic disorders was conducted. RESULTS: Tic disorders are characterized by repetitive and often disabling movements that primarily involve the eyes, face, neck, and voice apparatus. In addition, patients with tics may have eye problems related to medications used to control tics. Some eye signs are atypical in tic disorders and their presence may suggest alternate diagnoses. CONCLUSION: Eye tics can be disabling and a optometrist should be familiar with ocular manifestations of tics and ocular dysfunction related to tic treatment, and also be aware of other neurologic conditions confused with tics. PMID- 9409104 TI - Professional behavior and the optometric profession. AB - BACKGROUND: Optometry has been recognized as a profession within the United States, both legally and socially, for the better part of the past century. Historically, there have been expectations placed on the behavior of individuals within the professions that would not generally be placed on the general business person. These expectations have existed to protect the clients or patients of professionals from incompetence, uncaring, or selfish excesses. Behavior of an exceptional nature is expected of professionals because of the unusual vulnerability of clients and patients to unprofessional conduct. Doctors of Optometry, as members of the optometric profession, have professional standards placed on their behavior. METHODS: A search of the literature was conducted to discover the historical and current bases for setting standards for professional behavior. RESULTS: The literature search reinforces the rationale for the optometric professions long-standing practice of setting standards for professional conduct. CONCLUSIONS: Individual Doctors of Optometry will find that the trust resulting from high standards of professional conduct bring many positive benefits to the doctor-patient relationship. The rewards that come to both the doctor and patient from these trusting relationships make the practice of optometry truly an exceptional experience. PMID- 9409106 TI - Efficacy and durability of ultraviolet tints in CR-39 ophthalmic lenses. AB - BACKGROUND: Ocular protection from solar ultraviolet (UV) radiation has been emphasized in recent years as a result of the thinning of the ozone layer in the atmosphere. The purpose of this study was to evaluate the absorptive properties of UV tints in CR-39 lenses. METHODS: We used a spectrophotometer to measure the UV transmittance of three groups of UV tinted CR-39 lenses, including (1) lenses tinted by local optical laboratories: (2) lenses tinted by us, using commercially available dyes: and (3) stock UV lenses that have UV absorptive molecules throughout the lens. We also tested the durability of these tints to daily washing/drying by measuring their UV transmittance characteristics at 3, 6, and 12 months. RESULTS: All the tested lenses absorbed all of the UV-B and at least 99% of UV-A. The durability of these UV tints when exposed to daily washing/drying was excellent: all lenses continued to absorb all of the UV-B and at least 99% of UV-A after 1 year. CONCLUSIONS: These data suggest that UV tinted CR-39 lenses provide protection against UV radiation that meets the ANSI Z80.3 1996 Standard for non-prescription sunglasses and fashion eyewear. Furthermore, normal daily washing/drying for 1 year does not cause a significant decrease in the protective effect of the UV tint. PMID- 9409105 TI - Vision profile of the athletes of the 1995 Special Olympics World Summer Games. AB - BACKGROUND: Special Olympics were organized in 1968 by Eunice Shriver as a program of physical fitness for individuals with mental handicaps. Numerous epidemiologic studies report an increase in visual problems in this population. METHODS: A comprehensive vision screening was conducted at the 1995 Special Olympic World Summer Games to identify the prevalence of visual anomalies in this select group. Testing included visual acuity, refractive error, ocular motor skills, stereopsis, color vision, contrast sensitivity, eye-hand coordination, eye-foot coordination, and an ocular health evaluation. RESULTS: Nine hundred five special athletes, ranging in age from 8 to 58 years, participated in the screening. More than 65% of the participants had not received eye care for more than 3 years. The most commonly reported symptom was difficulty in seeing. Other ocular health problems included refractive errors, poor distance monocular acuity, and strabismus. CONCLUSION: The results of the study indicate that Special Olympians have a high prevalence of vision anomalies that may go undetected. This population demonstrates a high prevalence of refractive errors, decreased visual acuity, ocular health problems, and strabismus. In addition, based on the number of individuals who have not received eye care within the last year, there is a need to increase the availability of vision care to these and other people with mental retardation. PMID- 9409107 TI - Differential diagnosis in facial nerve palsy: a clinical review. AB - BACKGROUND: The facial nerve possesses five functional components and manifests a complex course from its origin in the motor cortex to its peripheral distribution. Pathologies that impact the facial nerve in various locations along its route result in characteristic clinical manifestations that often involve other neurologic entities. CASE REPORTS: Case reports of three patients who manifested lesions of the facial nerve are presented. Each case represents a specific facial nerve pathology occurring within the supranuclear, nuclear, and infranuclear location. An anatomic, regional, and etiologic approach to the spectrum of facial nerve disorders is provided. Additionally, hyperkinetic facial disorders is discussed, and the management of facial nerve palsy is emphasized. CONCLUSION: The clinician must understand the fundamental anatomy and distribution of the facial nerve in order to localize lesions and institute the appropriate management. Abnormalities of lid position and insufficient corneal wetting are problematic. All efforts should be directed toward the maintenance of corneal integrity by appropriate wetting strategies. PMID- 9409108 TI - Paraocular sinus mucoceles. AB - BACKGROUND: Patients with paraocular sinus masses may manifest ocular complications, including orbital displacement, proptosis, diplopia, restricted extraocular muscles, decreased vision, chemosis, pain, and optic neuritis. METHODS: Two patients with paraocular sinus masses came to our clinic for examination. One had proptosis and orbital displacement and a chief symptom of increasing diplopia, along with dull brow pain and sudden decreased vision of the left eye. He was diagnosed with polypoid disease and mucoceles of the ethmoidal and frontal sinuses. The second patient experienced pain and swelling under his left eye. He had an upper respiratory tract infection with sinusitis, which in turn increased the size of the sinus cyst. RESULTS: Computed tomography (CT) of the sinuses and orbits was ordered for both patients and confirmed the presence of mucoceles in each case. The first patient's signs and symptoms decreased over 2 weeks, so surgical intervention at that time was not necessary. The second patient was treated with 60-mg pseudoephedrine q.i.d. and 250-mg amoxicillin/potassium clavulanate q.i.d. CONCLUSIONS: Mucoceles are slow-growing polyp-like cysts of the sinuses; these cysts may be sterile in composition or harbor purulent infection (mucopyocele). In either case; they are space-occupying lesions that increase in size as mucus secretions continue, and can be exacerbated by active sinusitis. Differential diagnosis includes thyroid eye disease, orbital pseudotumor, infection, trauma, benign or malignant tumors, encephalocele, or meningiocele. PMID- 9409109 TI - Retinopathy secondary to anemia from myeloid metaplasia in polycythemia vera. AB - BACKGROUND: Polycythemia vera is a chronic clonal disorder associated with excessive proliferation of erythrocytes, leukocytes, and thrombocytes, as well as an accompanying splenomegaly. Ocular manifestations of polycythemia vera include occipital cortex transient ischemic attacks, transient monocular blindness, vaso occlusive disease, and retinal hemorrhages. CASE REPORT: A 56-year old man with longstanding polycythemia vera sought treatment for a chief symptom of blurred vision in the left eye and a red tinge to things first noticed on awakening that morning. He had preretinal and intraretinal hemorrhages and was subsequently found to be severely anemic as a result of postpolycythemic myeloid metaplasia after years of phlebotomy. Splenectomy controlled his anemia and thrombocytopenia, allowing transient improvement of the retinal hemorrhages. Acute leukemia subsequently developed and the patient died 7 weeks after initial examination. CONCLUSION: In this case, preretinal and intraretinal hemorrhages were found in a patient with longstanding polycythemia vera. The exact origin of these hemorrhages is uncertain. They are probably secondary to anemia, but the possibility that they are sites of extramedullary hematopoiesis must be considered. The appearance of retinal hemorrhages warrants careful investigation to rule out diabetes, hypertension, and anemia, as well as the various other blood dyscrasias. PMID- 9409111 TI - Nationwide poll on patient safety--100 million Americans see medical mistakes directly touching them. PMID- 9409110 TI - Effective stroke prevention in atrial fibrillation requires close laboratory monitoring--INR consensus statement. PMID- 9409112 TI - A treatise on Hansen's disease (leprosy) in Hawaii in the 1800s. PMID- 9409113 TI - Treatment of catheter-induced obstruction of the superior vena cava with Wallstent endoprosthesis. PMID- 9409114 TI - Aortic involvement in Marfan's syndrome. PMID- 9409115 TI - Radiological case of the month. Renal angiomyolipoma with perinephric hemorrhage. PMID- 9409116 TI - Photodynamic therapy: applications in bladder cancer and other malignancies. AB - Photodynamic therapy (PDT) has gained popularity in the past 10 years because of advances in laser and pharmacokinetic technologies and the development of new photosensitizers. Early studies on PDT with focal illumination for papillary bladder cancer obtained reasonable response rates for small tumors but recurrence was common. Whole bladder irradiation, once a suitable light-delivery system had been developed, gave promising outcomes with acceptable rates of complications. PDT for prostate cancer is still at the experimental stage but initial results have been promising. Clinical trials of PDT for brain tumors have shown no significant complications but no improvement in survival rate compared with other treatment modalities. PDT is particularly useful for early superficial lung cancers that are localized to one or a few discrete sites; it is also safe to use in patients who are too sick to be treated with conventional therapies. Preoperative PDT has reduced the extent of surgery necessary in some patients. Clinical experience with PDT for gynecological cancer is limited and prospective studies are needed. In head and neck oncology, PDT should prove a useful option, but methodological problems need to be overcome. Good responses of esophageal cancer to PDT have led to governmental approval of Photofrin, a photosensitizer, in several countries for either palliative use or treatment of inoperable or recurrent cancer. The use of PDT for early gastric cancer has great potential but several technical problems remain. PDT has proven generally effective for skin cancer when hematoporphyrin derivative or Photofrin is used but more long-term follow-up data are required for PDT with 5-aminolevulinic acid. Overall, PDT is changing from a scientific curiousity into an accepted modality for the treatment of cancer, with an improved likelihood of finding further clinical applications. PMID- 9409117 TI - Predicting the outcome of hemodialysis arteriovenous fistulae using duplex ultrasonography. AB - Duplex ultrasonography is a reliable method for assessing the anatomic features and blood flow rate of a vascular access point of hemodialysis. We assessed the value of measurement of cross-sectional area and blood flow rate of the major outflow veins using duplex ultrasonography in predicting the outcome of fistulae. Radiocephalic arteriovenous fistulae were created in 126 consecutive end-stage renal failure patients (55 men, 71 women; aged 20-83 yr) and examined using duplex ultrasonography in the second week following surgery. Examinations were repeated in 45 of the 126 fistulae in the third week. The outcome of new fistulae was classified as success or failure. The failure group fistulae were further classified as delayed maturation or primary failure. The cross-sectional area (12.1 +/- 3.5 vs 6.9 +/- 2.4 mm2) and blood flow rate (825.6 +/- 424.3 vs 303.7 +/- 114.5 mL/min) were significantly lower in the failure group, but there was no difference between the subgroups of primary failure and delayed maturation. Receiver-operating characteristic plots were generated for cross-sectional area and blood flow rate. The best cut-off point for distinguishing successful outcome from failure was 8.5 mm2 for cross-sectional area (sensitivity 0.823, specificity 0.867, positive predictive value 0.952, negative predictive value 0.605) and 425 mL/min for blood flow rate (sensitivity 0.813, specificity 0.933, positive predictive value 0.975, negative predictive value 0.609). Our findings show the cross-sectional area and blood flow rate, as measured using duplex ultrasonography, are useful in predicting the outcome of vascular access points of hemodialysis. PMID- 9409118 TI - A multidisciplinary pulmonary rehabilitation program for patients with moderately severe chronic obstructive pulmonary disease. AB - A multidisciplinary pulmonary rehabilitation program was conducted for 13 outpatients (mean age 66 +/- 6.7 yr) with moderately severe chronic obstructive pulmonary disease. Changes in pulmonary function and blood gas data were not significant. Exercise capability, including 6-minute walking distance (WkD6), maximal work load (WkLmax), endurance time, and maximum heart rate, improved significantly (p < 0.05), as did subjective symptoms and quality of life. Of the observed changes, only baseline PaO2 and oxygen saturation were positively correlated with changes in maximum heart rate. The initial maximum heart rate was inversely related to both the absolute and percentage improvement. There were no significant relationships between improvement in WkD6 and age, initial arterial blood gas, or pulmonary function, but a significant relationship was found between baseline forced expired volume in the first second (FEV1) and percentage change in WkLmax. Our results indicate that patients with moderately severe chronic obstructive pulmonary disease can improve their exercise capacity, subjective symptoms, and quality of life through a pulmonary rehabilitation program. All patients can increase their endurance, regardless of their initial exercise performance. Maximum heart rate and FEV1 are predictors of exercise capability improvement. PMID- 9409119 TI - Twenty-four-hour ambulatory esophageal pH monitoring in patients with symptoms of gastroesophageal reflux. AB - We investigated the relationship between gastric acid reflux patterns and esophageal mucosal damage in 142 patients with symptoms suggestive of gastroesophageal reflux and esophagitis. Each patient underwent endoscopy of the esophagus and 24-hour ambulatory esophageal pH monitoring. Endoscopy revealed varying degrees of esophagitis in 37 patients and a normal esophagus 105 patients. No significant difference in symptoms was found between patients with and without esophagitis. Sixteen of the 37 patients with esophagitis had a normal composite score on pH monitoring, whereas 24 of the 105 patients without esophagitis had an abnormal composite score (p < 0.001). Comparison of individual pH parameters during the study showed more reflux episodes (pH < 4), a higher percentage of reflux time, and more reflux episodes longer than 5 minutes in the group with esophagitis than in the without (p < 0.05). Both the percentage of reflux time and the number of reflux episodes longer than 5 minutes/hour were significantly higher (p < 0.05) in the group with endoscopically proven esophagitis in the supine period, but not in the upright period. There was a positive correlation between the severity of endoscopic esophagitis and the percentage of reflux time in the supine period (r = 0.64; p < 0.001), and the number of reflux episodes longer than 5 minutes/hour in the supine period (r = 0.56; p < 0.001). In conclusion, our patients with peptic esophagitis had significant gastric acid reflux and impaired acid clearance by the esophagus in the supine position. We advocate that 24-hour ambulatory esophageal pH monitoring should be considered in Chinese patients with normal endoscopy findings but with symptoms suggestive of gastroesophageal reflux disease. PMID- 9409120 TI - Diagnostic role of endoscopy, stool culture, and toxin A in Clostridium difficile associated disease. AB - This retrospective study was designed to assess the roles of stool culture for Clostridium difficile, detection of the presence of toxin A, and endoscopic examination in the diagnosis of Clostridium difficile-associated disease (CDAD). From January 1994 through September 1996, there were 213 patients with stool cultures positive for C. difficile in National Taiwan University Hospital. Of these, 126 had CDAD. There were 87 asymptomatic carriers of C. difficile in our study, 12 of whom were positive for toxin A. In addition, seven patients with pseudomembranous colitis (PMC), who were either culture-negative or not tested, were included in the study. The positive predictive values of stool cultures for CDAD and PMC were 59% and 32%, respectively. The positive predictive values of toxin A for CDAD and PMC were 41% and 43%, respectively. Seventy-eight patients (59%) improved with supportive treatment after discontinuing antibiotics. We concluded that stool culture for C. difficile and discontinuation of antibiotics should be the standard approach for patients with suspected CDAD. Endoscopic studies can eliminate some other possible causes of diarrhea such as inflammatory bowel disease, allow biopsies of suspicious lesions, and reveal the severity of CDAD. Toxin assay results need to be interpreted together with the clinical data. PMID- 9409121 TI - Outbreak and control of a rotaviral infection in a nursery. AB - Neonatal rotaviral infection generally causes an asymptomatic or mild illness. Once introduced into a nursery, it is very difficult to eradicate. We prospectively studied an outbreak of rotavirus infection in a normal newborn nursery from October 1994 through May 1995. Stool samples from infants more than 24 hours old were tested for rotaviral infection, either weekly, biweekly, or monthly. Rotavirus was identified in 164 (16%) of 1,037 tested neonates. Ninety four (57%) rotavirus-positive neonates became symptomatic: 56 had diarrhea, 26 developed fever (rectal temperature > 38 degrees C), 25 experienced vomiting, 17 showed poor feeding, and 14 had an elevated core temperature. In total, 24 neonates were evaluated for suspected sepsis. RNA electropherotyping of samples from 91 neonates revealed infection by the same rotavirus strain in all cases. This strain differed from that isolated from 64 rotavirus-infected infants and toddlers in the pediatric ward during the same period. Infection control procedures (hand washing, isolation of infected neonates, and careful management of diapers) and early discharge of uninfected neonates were instituted, and the outbreak was eradicated 8 months after the onset. Our findings indicate that many rotavirus-infected term neonates become symptomatic and have signs suggestive of sepsis. Extended hospital stay may be an important factor in promoting rotaviral transmission. Thus, early discharge may be an additional effective method of controlling rotavirus outbreaks in a nursery. PMID- 9409122 TI - Initial drug resistance of Mycobacterium tuberculosis in Taiwan. AB - The prevalence and mortality rate of pulmonary tuberculosis in adults are high in Taiwan. Because the emergence of drug-resistant tuberculosis is one of the major causes of this sustained high tuberculosis mortality, surveillance of initial drug resistance is important. We tested Mycobacterium tuberculosis isolates from 1,935 newly diagnosed tuberculosis patients from January 1990 through December 1995 at the Taiwan Provincial Chronic Disease Control Bureau. The overall initial drug resistance rate was 12.3%; 8.7% of isolates were resistant to only one drug, 2.6% to two drugs, 0.7% to three drugs, and 0.3% to four drugs. The resistance rates to individual drugs were: streptomycin, 5.7%; isoniazid, 9.2%; ethambutol, 0.7%; and rifampin, 1.5%. The frequency of multidrug-resistant M. tuberculosis (resistant to at least isoniazid and rifampin) was 1.2%. In view of the high initial isoniazid resistance rate and low initial ethambutol resistance rate, ethambutol should be added to the regimen for the initial treatment of tuberculosis in Taiwan. The emergence of multidrug-resistant M. tuberculosis is ominous and should be considered when treating patients in Taiwan. PMID- 9409123 TI - Clinical features and outcome of Acanthamoeba keratitis. AB - We retrospectively reviewed 11 patients with culture-proven Acanthamoeba keratitis who presented at the National Taiwan University Hospital between 1989 and 1996. We assessed predisposing factors, initial diagnosis, clinical presentation, treatment, and outcome. A history of contact lens-wear, poor contact lens hygiene, intractable eye pain, and ring infiltrates in the cornea were the most prominent characteristics and clinical manifestations. Acanthamoeba keratitis was often misdiagnosed, with herpetic keratitis (7/11) being the most common initial diagnosis from referring hospitals. These patients were usually treated on the basis of the inaccurate diagnosis for more than 1 month (range 1-8 mo) before referral. All patients ultimately received penetrating keratoplasty because of poor response to delayed medical treatment. We suggest that inadequate contact lens hygiene may be important in Acanthamoeba keratitis. This condition is often misdiagnosed and, as early diagnosis is a major factor for successful medical treatment in such patients, awareness in clinical practice is critical. PMID- 9409124 TI - Macrodactyly of the feet and hands. AB - We reviewed the records of 16 patients with true macrodactyly and analyzed the typical clinical features and methods of treatment. Fourteen feet were involved in 13 patients (one was bilaterally affected). Three hands were involved in three patients. Clinically, all lesions in the hands and lesions in 11 of 14 feet involved the preaxial side. There was multiple digit involvement in two hands and 11 feet. Progressive macrodactyly (10 feet and two hands) was more common than the static type (four feet and one hand). Proximal involvement of the sole or palm occurred in seven feet and one hand; all cases were of progressive macrodactyly. Enlargement of the metatarsals or the metacarpals was frequent (11 feet and two hands). The growth behavior and extent of bony involvement were similar in patients with hand involvement and those with foot involvement. Fourteen patients had additional clinodactyly, either medial or lateral. The toes of eight feet had angular deformities in the sagittal plane; most were angulated dorsally. Nine patients underwent surgery and two had repeated surgery. The reduction procedures included debulking, ray resection, toe resection, phalangeal resection, and phalangeal epiphysiodesis; the corrective procedures included wedge osteotomy, interdigitalization, and split thickness skin graft. Of the nine patients surgically treated, five had good results and four had fair results. Of the seven patients without surgical repair, three had fair results and four had poor results. Surgical debulking, phalangeal resection, ray resection, and phalangeal epiphysiodesis produced significant improvement in macrodactyly of the feet and hands. Toe resection was not as beneficial. PMID- 9409125 TI - Biometric study of acute primary angle-closure glaucoma. AB - We investigated the biometric characteristics of acute primary angle-closure glaucoma (PACG) patients and sought guidelines to predict subjects at high risk of acute attack. We enrolled 80 patients who suffered acute attack of angle closure glaucoma and 60 nonglaucoma subjects. The biometric data, measured with A scan ultrasonography, and the outcomes after various treatments were analyzed. Six patients were excluded because they were lost to follow-up. In the acute PACG patients, the axial length (mean +/- standard deviation) was 22.25 +/- 0.77 mm, the anterior chamber depth (corneal thickness included) was 2.28 +/- 0.23 mm, and the lens thickness was 4.94 +/- 0.28 mm. In the nonglaucoma subjects, the axial length was 23.26 +/- 0.76 mm, the anterior chamber depth (corneal thickness included) was 3.11 +/- 0.25 mm, and the lens thickness was 4.48 +/- 0.30 mm. An anterior chamber depth of less than 2.70 mm was the most sensitive (94%) and specific (94%) parameter to differentiate acute PACG patients from nonglaucoma subjects. The intraocular pressure was controlled within the satisfactory range in 58 patients by laser iridectomy, with or without antiglaucoma medication. Eight patients needed additional gonioplasty and eight required filtering surgery to control the intraocular pressure. Our findings provide useful information about the biometric data of eyes of patients with acute angle-closure glaucoma and offer a useful guide to predict those patients at high risk of acute attack. PMID- 9409126 TI - Osteomyelitis and tenosynovitis due to Mycobacterium marinum in a fish dealer. AB - Osteomyelitis caused by nontuberculous mycobacteria is rarely reported. We describe a case of tenosynovitis and osteomyelitis of the right middle finger and metacarpal bone caused by Mycobacterium marinum in a fish dealer. This 52-year old woman suffered progressive pain, numbness, tenderness, and erythematous swelling of the right middle finger over a 2-month period. A radiograph of the right hand disclosed osteolytic lesions at the third metacarpal bone and the third proximal phalanx. She was treated successfully with repeated surgical debridement and antimicrobial agents, including clarithromycin, ethambutol, rifampin, and doxycycline for 1 month, followed by ethambutol and clarithromycin. Pathologic examination of the debrided tissue disclosed epithelioid granuloma, caseous necrosis, and numerous acid-fast bacilli, which were later identified as M. marinum using conventional biochemical tests and by the characteristic gas liquid chromatogram of esterified cellular fatty acid. The wound healed completely after 7 months of treatment. The patient is still under treatment, and clarithromycin and ethambutol will be given for a total of 18 months. PMID- 9409127 TI - Systemic sclerosis with pulmonary involvement and right ventricular failure in a child. AB - We report a very rare case of systemic sclerosis in a 6-year-old girl. She presented with diffuse scleroderma, Raynaud's phenomenon, pulmonary interstitial fibrosis, pulmonary hypertension, and right ventricular failure. The diagnosis was confirmed by skin manifestations, high resolution computed tomography, cardiac catheterization, and anti-nuclear antibodies. Nifedipine, prednisolone, digoxin, and furosemide were given. There was remission of the right ventricular failure and dyspnea, and the skin showed partial improvement. The patient remained asymptomatic for a year. The symptoms of respiratory and right heart failure developed again after an episode of lower respiratory tract infection and she eventually died. We discuss the clinical manifestations, treatment, and outcome. PMID- 9409128 TI - Trifascicular block with intermittent complete atrioventricular block in a child. AB - Trifascicular block, which consists of impaired conduction in the three main fascicles of the ventricular conduction system, may progress to high-grade or complete atrioventricular block. It rarely occurs in children with a structurally normal heart. We report a case of trifascicular block in a previously healthy 9 year-old girl. The patient presented with repetitive seizures. The electrocardiogram showed a complete right bundle branch block, left axis deviation, PR interval prolongation, and intermittent high-degree and complete atrioventricular block. She was successfully treated with a temporary pacemaker and her electrocardiogram returned to a normal sinus rhythm in 3 days. She has remained well over a 4-year follow-up. Although her cardiac enzyme levels were normal, the clinical course and electrocardiography findings suggested myocarditis. We emphasize the diagnosis of trifascicular block using electrocardiography; clinical outcome is good, if the patient is managed properly. PMID- 9409129 TI - Legislative approaches to the regulation of the chiropractic profession. AB - Traditional and complementary health care services have a growing and significant role in both developed and developing countries. In the United Kingdom the British Medical Association (BMA) has identified five complementary approaches to health care that should now be regarded as "discrete clinical disciplines" because they have "established foundations of training and have the potential for greatest use alongside orthodox medical care". These are acupuncture, chiropractic, herbalism, homeopathy and osteopathy. The BMA recommended that there should be legislation to regulate these disciplines and the Chiropractors' Act enacted in the U.K in 1994. The chiropractic profession was founded in the United States in 1895, and the practice of chiropractic has been regulated in the United States and Canada since the 1920s, in Australia since the late 1940s, in New Zealand and South Africa since the 1960s, and more recently in Asia, Europe, Latin America and elsewhere. Figure 1 lists the countries which currently recognize and regulate the chiropractic profession. Many countries, such as Japan with approximately 10,000 chiropractors with different levels of education, and Trinidad & Tobago with 5 chiropractors who are graduates of accredited chiropractic colleges in North America, are considering legislation. Croatia, with 3 chiropractors, is preparing legislation. Cyprus, with 6 chiropractors, has legislation. Even in countries such as these, where the profession is small, there are compelling public interest arguments for regulation. This is especially true in the 1990s. One reason is the growing incentive for lay healers and others without formal training to use the title "chiropractor" as chiropractic practice gains increasing acceptance. The majority of chiropractic practice involves patients with non- specific or mechanical back and neck pain. The chiropractic approach to management, which includes spinal adjustment or manipulation, other physical treatments, postural advice, rehabilitative exercises and early return to activities, formally only had empirical evidence of success. Now there is firm scientific support. Recent national, evidence- based, multi-disciplinary guidelines in Canada (neck pain), the United Kingdom (back pain), and the United States (back pain) support these methods as a first line of management for most patients. Another reason for regulation is that international standards of chiropractic education and scope of practice have been established by appropriate chiropractic organizations, including the World Federation of Chiropractic which represents national associations of chiropractors in 63 countries. This paper now reviews current legislation worldwide. PMID- 9409130 TI - Some observations on post-coma unawareness patients and on other forms of unconscious patients: policy proposals. AB - The concern of this paper is with various forms of unconsciousness. Special attention will be given to patients in PCU commonly termed as Persistent Vegetative State (PVS). I reject the term PVS, arguing that for ethical reasons we should strive to resort to terminology that does not offend the patients and their loved ones. I further urge hospitals not to cease treatment of PCU patients younger than 50 years old within a period of less than two years. This is especially true for patients who entered a state of unawareness due to traumatic causes. The two-years waiting period should be regarded as the minimum period of evaluation before foregoing hopes for patients' rehabilitation and return to some form of cognition. I provide data and human stories from the Israeli experience to substantiate this argument. PMID- 9409131 TI - The Ombudsman's involvement in ensuring patients' rights. AB - An analysis has been carried out on the extent and character of patient complaints regarding physicians' conduct, received by the Danish Parliament's Ombudsman during the period 1983-1993. Only a few of the complaints to the Ombudsman are treated completely or partially, because the Ombudsman only examines the administrative handling of the case, and declines to consider the medical opinion. The consequences of the actions chosen by the Ombudsman, regarding ensuring patients' rights, are discussed. In the light of the establishment of more local patient ombudsman systems, there is an ongoing discussion as to just how effective these will be to ensure the legal rights of patients. The possibility of establishing a national patient ombudsman is also discussed. PMID- 9409132 TI - Pseudopsychopathic schizophrenia--a neglected diagnostic entity with legal implications. AB - Pseudopsychopathic schizophrenia (PPS) forms a diagnostic unity, comprising aspects of schizophrenic process and antisocial personality disorder. Perusal of the psychiatric literature reveals that this diagnostic entity has fallen into oblivion. Recognition of this neglected category is important from therapeutic, academic and medico-legal points of view. PMID- 9409133 TI - Practices and attitudes among Swedish psychiatrists regarding the ethics of compulsory treatment. PMID- 9409134 TI - Mental illness in India and Britain: theory and practice. AB - The paper argues that conceptions of mental illness and its treatment often stem from the normative social and cultural constructions of mental illness. Psychiatrists, psychologists, and other health professionals do not work in a social vacuum; they work within the accepted traditions and values prevalent in their culture. To understand mental illness it is therefore necessary to examine the salient normative beliefs, attitudes, and values of a given culture. The paper proposes a cross-cultural theoretical and empirical model which permits a close examination and comparison of mental illness in two cultures: India and Britain. The proposed model from which several testable hypotheses have been deduced, rests on the following four factors; [Table See Text]. The nature and the importance of the factors in explaining mental illness and the culture specific treatment strategies which follow in the two cultures are critically discussed in the paper. PMID- 9409135 TI - Whistle-blowing knowledge and attitude of Israeli nurses. PMID- 9409136 TI - Genetics, human rights and employment. American and European perspectives. AB - The United States of America and the European Countries have responded quite differently to the to the human rights of candidate workers as imposed by genetic and predictive health testing. Whereas the United States traditionally relies largely on the non-discrimination principle, the European countries seem to attach more value to the right to privacy and the principle of social justice. A recently published communication of the Commission of the European Communities seems to bridge these two legal traditions. PMID- 9409137 TI - Consent in the 90's. AB - Patients consent to surgical procedures is a universal issue in medical law. The legal position in Australia, because it falls somewhere between the North American doctrine of "informed consent", and the Bolam test in the United Kingdom, will be of interest to all clinicians and legal practitioners dealing with these issues. The High Court of Australia's decision in Rogers v Whitaker (1992) 109 ALR 625 is now the leading decision regarding of consent to surgical procedures in Australia. The author draws a thumbnail sketch of the law in Australia, and discusses some loopholes in the legal structure established by the Australian High Court. The author supports the view taken by the High Court of Australia, and provides some material to contest the allegation made by clinicians that lawyers in Australia are in some way responsible to "opening the floodgates" for an increase in legal claims by patients. PMID- 9409138 TI - Medical disclosure and consent forms: proposal for an international standard. AB - Disclosure of material risks prior to invasive medical and surgical procedures remains a fiction derived from legal idealism rather than a clinical reality. Where disclosure has been inadequate, legal allocation of individual medical blame fails to address ongoing systems problems in the disclosure process and thereby potentially jeopardizes patient safety. Patient signature of disclosure and consent forms is a standardised international ritual with great potential for ensuring that international ideals and principles of medical ethics (such as those relating to disclosure of risk) become a recognised and consistent part of global medical practice. This article encourages the development of an international standard for disclosure and consent forms. PMID- 9409139 TI - Deaths and medical attention in police custody. AB - This article presents data from a study on the medical and monitoring procedures surrounding all 59 deaths in Dutch police stations in the period 1983-1993. In many cases, the medical procedures and monitoring procedures were not in accordance with the Dutch ministerial guidelines. Notification of physicians, transference to hospitals, medical examinations, and monitoring procedures showed several shortcomings. Based on the findings, several measures for the prevention of deaths in police custody are discussed. PMID- 9409140 TI - Outcomes of regular vs. extended alcohol/drug outpatient treatment: I. Relapse, aftercare, and treatment re-entry. AB - Alcohol and drug patients were randomized into two groups, one receiving three months and the other six months of outpatient treatment to determine differences in treatment outcomes. Most clients had received prior 30 days of inpatient treatment. Patients were contacted after the first 70 days of outpatient treatment and 12 refused participation. Consenters were randomized and assigned into control (n = 103) and experimental (n = 127) groups, and interviewed at discharge, and three and six months later. A gratuity of $10.00 was offered after a completed phone interview. Data were analyzed using chi-square, t-test, and multivariate logistic regression techniques. Controls had lower treatment drop out and higher follow-up attrition rates than experimentals. There were no major differences in reported subsequent alcohol/drug use, or attendance to aftercare, Alcohol Anonymous (AA) and support groups during the 3 and 6 months follow-up surveys. More controls re-entered treatment than experimentals at 3 months post treatment, but there was no such difference at 6 months post-treatment. In terms of ancillary effects, experimentals had slightly more desirable outcomes with respect to abstinence at time of discharge, and use of cocaine at 3 months follow up. Controls were more likely to use cocaine and less likely to re-enter inpatient treatment or attend aftercare than experimentals. At six months the few who reported using painkillers were controls. Relapse was predictably influenced both at 3 and 6 months by pretreatment use of cocaine as primary drug, and by duration of abstinence from all chemicals. The predictive influence of cocaine was greater at 3 than at 6 months post-discharge. PMID- 9409141 TI - Artificial wombs: medical miracle, legal nightmare. AB - The fact that the development of the artificial uterus is nearing completion is a mixed blessing. It will provide numerous benefits within the field of pediatric medicine, such as ensuring normal development to term of extremely premature foetuses, but it will pose numerous problems which the law is ill-equipped to handle. The legislature will have to examine the current definition of 'parent' which is based on normal conception practices. It will also have to determine whether only married couples should have access to this technology and under which conditions. PMID- 9409142 TI - A judicial point of view with regard to the testimony of medical experts. AB - This is an era of the expert witness. The classic description of an expert--"... no scientific man ought ever to become the partisan of a side: he may be the partisan of an opinion in his own science"--Lord MacMillan. The current tendency to be less than impartial or objective: the anxieties of Lord Woolf in Access to Justice-Factors which may lead to partiality: (a) explosions of claims for personal injury or product liability; (b) the emergence of the contingency fee as the primary fund for such litigation. Problems facing an expert--the arrogance of assurance--the power of presentation--the anxieties of the witness box. The virtues of education and training. The importance of judicial management to control expert evidence and avoid escalating costs and delay in litigation. The duty of the Judge is to ensure that the expert does not practice a fraud on the administration of justice. PMID- 9409143 TI - "No" to the intention/foresight distinction in medical end-of-life decisions. AB - The idea that the permissibility of an action (or an omission) depends on the agent's intention has played an important role in traditional moral thinking and the law and determines, at least in part, the permissibility of medical end-of life decisions. I argue that there are good reasons why the law ought not, in end of-life decisions, focus on the doctor's intention. The patient's interests or rights are a more appropriate focus. PMID- 9409144 TI - Informed consent--a ploy? PMID- 9409146 TI - Mating types and sexual development in filamentous ascomycetes. AB - The progress made in the molecular characterization of the mating types in several filamentous ascomycetes has allowed us to better understand their role in sexual development and has brought to light interesting biological problems. The mating types of Neurospora crassa, Podospora anserina, and Cochliobolus heterostrophus consist of unrelated and unique sequences containing one or several genes with multiple functions, related to sexuality or not, such as vegetative incompatibility in N. crassa. The presence of putative DNA binding domains in the proteins encoded by the mating-type (mat) genes suggests that they may be transcriptional factors. The mat genes play a role in cell-cell recognition at fertilization, probably by activating the genes responsible for the hormonal signal whose occurrence was previously demonstrated by physiological experiments. They also control recognition between nuclei at a later stage, when reproductive nuclei of each mating type which have divided in the common cytoplasm pair within the ascogenous hyphae. How self is distinguished from nonself at the nuclear level is not known. The finding that homothallic species, able to mate in the absence of a partner, contain both mating types in the same haploid genome has raised more issues than it has resolved. The instability of the mating type, in particular in Sclerotinia trifolorium and Botrytinia fuckeliana, is also unexplained. This diversity of mating systems, still more apparent if the yeasts and the basidiomycetes are taken into account, clearly shows that no single species can serve as a universal mating-type model. PMID- 9409147 TI - Cyclopropane ring formation in membrane lipids of bacteria. AB - It has been known for several decades that cyclopropane fatty acids (CFAs) occur in the phospholipids of many species of bacteria. CFAs are formed by the addition of a methylene group, derived from the methyl group of S-adenosylmethionine, across the carbon-carbon double bond of unsaturated fatty acids (UFAs). The C1 transfer does not involve free fatty acids or intermediates of phospholipid biosynthesis but, rather, mature phospholipid molecules already incorporated into membrane bilayers. Furthermore, CFAs are typically produced at the onset of the stationary phase in bacterial cultures. CFA formation can thus be considered a conditional, postsynthetic modification of bacterial membrane lipid bilayers. This modification is noteworthy in several respects. It is catalyzed by a soluble enzyme, although one of the substrates, the UFA double bond, is normally sequestered deep within the hydrophobic interior of the phospholipid bilayer. The enzyme, CFA synthase, discriminates between phospholipid vesicles containing only saturated fatty acids and those containing UFAs; it exhibits no affinity for vesicles of the former composition. These and other properties imply that topologically novel protein-lipid interactions occur in the biosynthesis of CFAs. The timing and extent of the UFA-to-CFA conversion in batch cultures and the widespread distribution of CFA synthesis among bacteria would seem to suggest an important physiological role for this phenomenon, yet its rationale remains unclear despite experimental tests of a variety of hypotheses. Manipulation of the CFA synthase of Escherichia coli by genetic methods has nevertheless provided valuable insight into the physiology of CFA formation. It has identified the CFA synthase gene as one of several rpoS-regulated genes of E. coli and has provided for the construction of strains in which proposed cellular functions of CFAs can be properly evaluated. Cloning and manipulation of the CFA synthase structural gene have also enabled this novel but extremely unstable enzyme to be purified and analyzed in molecular terms and have led to the identification of mechanistically related enzymes in clinically important bacterial pathogens. PMID- 9409145 TI - Arac/XylS family of transcriptional regulators. AB - The ArC/XylS family of prokaryotic positive transcriptional regulators includes more than 100 proteins and polypeptides derived from open reading frames translated from DNA sequences. Members of this family are widely distributed and have been found in the gamma subgroup of the proteobacteria, low- and high-G + C content gram-positive bacteria, and cyanobacteria. These proteins are defined by a profile that can be accessed from PROSITE PS01124. Members of the family are about 300 amino acids long and have three main regulatory functions in common: carbon metabolism, stress response, and pathogenesis. Multiple alignments of the proteins of the family define a conserved stretch of 99 amino acids usually located at the C-terminal region of the regulator and connected to a nonconserved region via a linker. The conserved stretch contains all the elements required to bind DNA target sequences and to activate transcription from cognate promoters. Secondary analysis of the conserved region suggests that it contains two potential alpha-helix-turn-alpha-helix DNA binding motifs. The first, and better fitting motif is supported by biochemical data, whereas existing biochemical data neither support nor refute the proposal that the second region possesses this structure. The phylogenetic relationship suggests that members of the family have recruited the nonconserved domain(s) into a series of existing domains involved in DNA recognition and transcription stimulation and that this recruited domain governs the role that the regulator carries out. For some regulators, it has been demonstrated that the nonconserved region contains the dimerization domain. For the regulators involved in carbon metabolism, the effector binding determinants are also in this region. Most regulators belonging to the AraC/XylS family recognize multiple binding sites in the regulated promoters. One of the motifs usually overlaps or is adjacent to the -35 region of the cognate promoters. Footprinting assays have suggested that these regulators protect a stretch of up to 20 bp in the target promoters, and multiple alignments of binding sites for a number of regulators have shown that the proteins recognize short motifs within the protected region. PMID- 9409148 TI - Rolling-circle replication of bacterial plasmids. AB - Many bacterial plasmids replicate by a rolling-circle (RC) mechanism. Their replication properties have many similarities to as well as significant differences from those of single-stranded DNA (ssDNA) coliphages, which also replicate by an RC mechanism. Studies on a large number of RC plasmids have revealed that they fall into several families based on homology in their initiator proteins and leading-strand origins. The leading-strand origins contain distinct sequences that are required for binding and nicking by the Rep proteins. Leading-strand origins also contain domains that are required for the initiation and termination of replication. RC plasmids generate ssDNA intermediates during replication, since their lagging-strand synthesis does not usually initiate until the leading strand has been almost fully synthesized. The leading- and lagging strand origins are distinct, and the displaced leading-strand DNA is converted to the double-stranded form by using solely the host proteins. The Rep proteins encoded by RC plasmids contain specific domains that are involved in their origin binding and nicking activities. The replication and copy number of RC plasmids, in general, are regulated at the level of synthesis of their Rep proteins, which are usually rate limiting for replication. Some RC Rep proteins are known to be inactivated after supporting one round of replication. A number of in vitro replication systems have been developed for RC plasmids and have provided insight into the mechanism of plasmid RC replication. PMID- 9409149 TI - Archaea and the prokaryote-to-eukaryote transition. AB - Since the late 1970s, determining the phylogenetic relationships among the contemporary domains of life, the Archaea (archaebacteria), Bacteria (eubacteria), and Eucarya (eukaryotes), has been central to the study of early cellular evolution. The two salient issues surrounding the universal tree of life are whether all three domains are monophyletic (i.e., all equivalent in taxanomic rank) and where the root of the universal tree lies. Evaluation of the status of the Archaea has become key to answering these questions. This review considers our cumulative knowledge about the Archaea in relationship to the Bacteria and Eucarya. Particular attention is paid to the recent use of molecular phylogenetic approaches to reconstructing the tree of life. In this regard, the phylogenetic analyses of more than 60 proteins are reviewed and presented in the context of their participation in major biochemical pathways. Although many gene trees are incongruent, the majority do suggest a sisterhood between Archaea and Eucarya. Altering this general pattern of gene evolution are two kinds of potential interdomain gene transferrals. One horizontal gene exchange might have involved the gram-positive Bacteria and the Archaea, while the other might have occurred between proteobacteria and eukaryotes and might have been mediated by endosymbiosis. PMID- 9409152 TI - Help! Managed care is killing my practice. PMID- 9409153 TI - Do no harm. Thalidomide, laser cavity preparation and oral hydrogen peroxide. PMID- 9409154 TI - Matter of life or death. PMID- 9409155 TI - Dismissed out of hand. PMID- 9409150 TI - Metabolism of sulfur amino acids in Saccharomyces cerevisiae. AB - Sulfur amino acid biosynthesis in Saccharomyces cerevisiae involves a large number of enzymes required for the de novo biosynthesis of methionine and cysteine and the recycling of organic sulfur metabolites. This review summarizes the details of these processes and analyzes the molecular data which have been acquired in this metabolic area. Sulfur biochemistry appears not to be unique through terrestrial life, and S. cerevisiae is one of the species of sulfate assimilatory organisms possessing a larger set of enzymes for sulfur metabolism. The review also deals with several enzyme deficiencies that lead to a nutritional requirement for organic sulfur, although they do not correspond to defects within the biosynthetic pathway. In S. cerevisiae, the sulfur amino acid biosynthetic pathway is tightly controlled: in response to an increase in the amount of intracellular S-adenosylmethionine (AdoMet), transcription of the coregulated genes is turned off. The second part of the review is devoted to the molecular mechanisms underlying this regulation. The coordinated response to AdoMet requires two cis-acting promoter elements. One centers on the sequence TCACGTG, which also constitutes a component of all S. cerevisiae centromeres. Situated upstream of the sulfur genes, this element is the binding site of a transcription activation complex consisting of a basic helix-loop-helix factor, Cbf1p, and two basic leucine zipper factors, Met4p and Met28p. Molecular studies have unraveled the specific functions for each subunit of the Cbf1p-Met4p-Met28p complex as well as the modalities of its assembly on the DNA. The Cbf1p-Met4p-Met28p complex contains only one transcription activation module, the Met4p subunit. Detailed mutational analysis of Met4p has elucidated its functional organization. In addition to its activation and bZIP domains, Met4p contains two regulatory domains, called the inhibitory region and the auxiliary domain. When the level of intracellular AdoMet increases, the transcription activation function of Met4 is prevented by Met30p, which binds to the Met4 inhibitory region. In addition to the Cbf1p-Met4p-Met28p complex, transcriptional regulation involves two zinc finger-containing proteins, Met31p and Met32p. The AdoMet-mediated control of the sulfur amino acid pathway illustrates the molecular strategies used by eucaryotic cells to couple gene expression to metabolic changes. PMID- 9409156 TI - Cortical bone resorption secondary to endodontic-implant pathology. A case report. AB - A single implant was inserted to replace the mandibular right lateral incisor in a 50-year-old Caucasian woman. After one month, an endodontic lesion formed on the adjacent canine, which went on to engulf the fixture. Even though endodontic therapy was instituted expeditiously on the canine, the implant exfoliated spontaneously the following month. There was a large radiolucency afterwards, suggesting resorption of both the labial and lingual cortical plates. A follow-up radiograph four months later revealed a residual detect along with delayed wound repair. PMID- 9409157 TI - Oral and maxillofacial surgery. A specialty altered by time and circumstance. AB - The evolution of oral and maxillofacial surgery into a specialty was aided by international strife in the first half of the century. Today the scope of OMS practice continues to expand as the result of an educational process that is responsive to the changing needs of the specialty. But understanding and acceptance of the OMS scope of practice lags behind. PMID- 9409158 TI - Long-term effects of political imprisonment: a group comparison study. AB - The study investigated the long-term effects of political imprisonment in the former German Democratic Republic. A group of non-treatment-seeking former political prisoners (n = 146) was compared with an age- and sex-matched group (n = 75). Assessments included the structured Diagnostic Interview for Psychiatric Disorders (German abbreviation: DIPS) for DSM-III-R/-IV diagnoses, a checklist of persecution and maltreatment, and other self-rated measures of post-traumatic stress disorder (PTSD), anxiety, depression, and dissociation. PTSD was assessed by the DIPS as current and lifetime diagnoses. Former political prisoners were imprisoned for 38 months on average. The former prisoners had a lower educational and lifetime occupational level than the comparison group. Results regarding diagnoses show a frequency of 30% current and 60% lifetime PTSD in the former prisoners group. Other anxiety disorders (e.g., claustrophobia, social phobia) outnumbered comorbid affective disorders. The level of dissociation was elevated in the former prisoners group. Intrusive recollections and hyperarousal were more common than avoidance/numbing symptoms. Despite differences in imprisonment duration between three historically defined eras of persecution, no differences appeared in the level of symptomatology. The results suggest that political imprisonment in the former German Democratic Republic had long-term psychological effects. Compared with an age- and sex-matched comparison group, the former political prisoners showed higher levels not only of post-traumatic symptomatology but also of other anxiety disorders and dissociation. PMID- 9409159 TI - Mortality after deliberate self-poisoning. A prospective follow-up study of 587 persons observed for 5279 person years: risk factors and causes of death. AB - A follow-up study of mortality and factors associated with death from various causes were done on two unselected groups of patients surviving deliberate self poisoning in 1978 and 1987. The persons were studied up to the end of 1993. In 1978 the group included 152 female and 101 male subjects and in 1987 the group included 190 female and 144 male subjects. By the end of 1993 a total of 37 (24%) of the female and 33 (33%) of the male patients admitted in 1978 had died (n.s.) and 18 (10%) of the female and 29 (20%) of the male patients admitted in 1987 had died (P < 0.01). The main causes of death were suicide and death from cardiovascular disease. The 5-year follow-up mortality more than doubled in males from 1978 to 1987 but decreased in females. In female subjects, the total follow up mortality was 3.6 times the expected ratio, with a 95% confidence interval (95% CI of 2.7-4.6); in male subjects it was 5.0 times the expected ratio (95% CI = 3.8-6.4). The cause-specific mortality ratio was highest for deaths from suicide--in the female group it was 65.5 (39.4-102.3) times the expected and in the male group 41.5 (26.0-62.8)--and from accidental poisoning--for females 50.0 (6.1-180.6) times the expected and for males 66.7 (24.5-145.1). In the female group none of the variables examined reached significance as predictors for subsequent suicide or death from unnatural causes. In the male group being aged 30 years or more came out as a predictor for subsequent suicide [relative risk (RR) = 5.66 (1.05-30.37)], while imprisonment came out as a protective factor [RR = 0.08 (0.01-0.64)]. Significant predictors for death from unnatural causes were: having been convicted (but not been in jail) [RR = 34.01 (1.07-1078.15)] and a serious suicidal intent [RR = 138.62 (1.38-13,946.79)]. It is concluded that patients who survive deliberate self-poisoning are at increased risk of death. The predictors for death are not very specific and are considered difficult to apply in the clinical work with these patients. PMID- 9409160 TI - A 3- to 6-year follow-up of former long-stay psychiatric patients in Northern Ireland. AB - Little is known about the first patients who left hospital before and during the official implementation of the hospital discharge policy in Northern Ireland. This study describes patterns of residential provision for former long-stay patients (approximately two-thirds of whom had an ICD-9 diagnosis of schizophrenia) discharged from the six major psychiatric hospitals in Northern Ireland between 1987 and 1990 (n = 321). It also employs several instruments within a retrospective survey design to examine outcomes for a 35% sample of people (112/321) discharged between 1997 and 1990 and followed up in 1993. Almost two-thirds (61%) had been discharged to independent living or low-staffed statutory settings. None of the group was homeless, one person was in prison and three people had committed suicide during the first 2 years after discharged. Almost one-third had to be re-admitted at some stage during the 6-year period and 13% had died. 'Moderate' to 'major problems' with most daily living skills were reported for less than 25% of people, while 15% or less had problem behaviour. Approximately 90% or more were satisfied with most aspects of their new homes and most also reported feeling happier (77%), healthier (63%) and more independent (78%) since discharge. However, social, recreational and occupational opportunities were limited. Purchasers, providers and practitioners need to review ways in which former long-stay patients might be empowered to live more meaningful and integrated lives in the community, particularly as the current government strategy for health and social well-being (1997-2002) in Northern Ireland points to the closure of existing psychiatric hospitals. PMID- 9409161 TI - Sexual and relationship problems amongst patients with severe chronic psychoses. AB - Recent studies have begun to examine the complexity and frequency of sexual and relationship problems amongst samples of community resident people with severe psychoses. For the purposes of this study, subjects with severe persistent psychoses and under the care of a single community team were interviewed using a semi-structured clinical interview and structured diagnostic interview of marital and sexual satisfaction. Functional disability was assessed by the Multnomah Community Ability Scale. Amongst those interviewed (n = 53) the prevalence of sexual difficulties was 47.5% and 30.8% for men (n = 40) and women (n = 13), respectively. The majority of men (82.5%) and some of the women (38.5%) were not in intimate relationships; 42.5% of men and 38.5% of women had never had a sexual relationship. Amongst community resident patients with severe psychoses, the level of unmet need for specific interventions (including assessment procedures, psychotherapeutic, pharmaco-therapeutic) for sexual and relationship dysfunction is high. This warrants evaluation of service structures and treatment packages tailored for this group. PMID- 9409162 TI - Belief systems of the general public concerning the appropriate treatments for mental disorders. AB - A study was conducted to assess the belief systems of the general public concerning the appropriate treatments for mental disorders and correlates of these belief systems. The study was based on the results of a household survey of the general public in Australia, using a national random sample of 2,031 adults aged 18-74 years. Respondents were given a vignette describing either a person with depression or one with schizophrenia, and were asked for their opinions about the helpfulness of various professional and non-professional treatments for the person described. A principal components analysis of the helpfulness ratings gave three factors: a Medical factor with high loadings on all drug treatments (except Vitamins) and on Psychiatric ward and ECT; a Psychological factor with high loadings on Counsellor, Social Worker, Phone counselling, Psychiatrist, Psychologist, Psychotherapy and Hypnosis; and a Lifestyle factor with high loadings on Close family, Close friends, Naturopath, Vitamins, Physical activity and Get out more. The same factors emerged from ratings of the two vignettes. Mean scores on scales constructed from the items with high loadings showed that the public tend to have a negative view of medical treatments and a positive view of psychological and lifestyle ones. However, medical treatments were rated more negatively for depression than for schizophrenia, psychological treatments were rated more positively for schizophrenia, and lifestyle treatments more positively for depression. Age, sex and education of respondents showed few associations with scores on the scales, although the better educated were more in favour of psychological treatments for both depression and schizophrenia and were less opposed to medical treatments for schizophrenia. Respondents who had suffered from the symptoms described in the schizophrenia vignette were more negative towards medical treatments. These findings about public belief systems could have implications for the provision of treatment: where there is a discrepancy in belief system between the patient and the clinician there may be poor adherence to treatment. PMID- 9409163 TI - The validity of the Arabic Edinburgh Postnatal Depression Scale. AB - For the purpose of this study, a consecutive sample of 95 postpartum women were assessed at 1 week postpartum with the (EPDS) and at 8 +/- 2 weeks postpartum using the Present State Examination (PSE). A moderate correlation between PSE total score and EPDS score was found (r = 0.57). A moderate agreement between EPDS and Catego diagnosis of depression was also found (Kappa = 0.52). Using a cut-off score of 12 on EPDS and Catego diagnosis as a criterion variable, the sensitivity and specificity of the scale were 73% and 90%, respectively. However, using a cut-off score of 10, the sensitivity of the scale rose to 91% without much fall in its specificity (84%). The internal reliability of the scale was 0.84 (alpha Cronbach). We conclude that the Arabic version of the EPDS is a reliable and valid screening tool for depression in postpartum women. PMID- 9409164 TI - Understanding childhood (problem) behaviors from a cultural perspective: comparison of problem behaviors and competencies in Turkish immigrant, Turkish and Dutch children. AB - Parents' reports of problem behaviors in 2,081 Dutch children, 3,127 Turkish children in Ankara and 833 Turkish immigrant children living in The Netherlands, aged 4-18 years, were compared. Dutch and Turkish versions of the Child Behavior Checklist (CBCL) were used. Immigrant children were scored higher than Dutch children on 6 of the 11 CBCL scales, most markedly on the Anxious/Depressed scale. Immigrant children were scored higher than Ankara children on five CBCL scales. However, these differences were much smaller than those found between immigrant and Dutch children. Furthermore, immigrant children's Total Problem scores did not differ from those for Ankara children. Turkish immigrant children have very similar patterns of parent-reported problem behaviors to children living in Turkey, although both groups of Turkish children showed higher levels of parent-reported problem behaviors than Dutch children. The higher scores for Turkish children on the Anxious/Depressed scale compared with their Dutch peers may be explained by cultural differences in parental perception of children's problem behaviors, as well as the threshold for reporting them, or by cultural differences in the prevalence of problems, for instance as the result of cross cultural differences in child-rearing practice. More research is needed to test the degree to which Turkish immigrant parents tend to preserve their cultural characteristics and child-rearing practices in Dutch society. PMID- 9409166 TI - Can managed care work in rural areas? PMID- 9409167 TI - Using asset protection strategies to shield your practice and savings from lawsuits. PMID- 9409165 TI - Social deprivation and psychiatric admission rates in Amsterdam. AB - The main subject of this study was the link between social indicators and the (re)admission rates for, and length of stay in, in-patient mental health care in Amsterdam. In a factor analysis of 15 sociodemographic variables, two principal components analysis factors were distinguished: housing quality and socioeconomic deprivation. The census variables and the factors almost all had high correlations with the crude admission rates as well as the rates standardised for age and sex. In general, the correlations with rates that were also standardised for marital status were significantly lower. This shows that many correlations between indicators and crude rates are determined to a significant extent by the marital status profile of an area. Socioeconomic deprivation is positively correlated with the proportion of readmissions and inversely correlated with average length of stay. PMID- 9409168 TI - The unfairness of the system. PMID- 9409169 TI - Hypercalcemia and pulmonary tuberculosis in east Tennessee. AB - In a study of 83 patients with active pulmonary tuberculosis who were treated in East Tennessee, only three developed hypercalcemia. The incidence of hypercalcemia in East Tennessee is markedly lower than that quoted in earlier studies performed in the United States. The explanation for the infrequent occurrence of elevated serum calcium in our population is probably multifactorial, but does not appear to be related to the selection of antituberculous agents. PMID- 9409170 TI - Infantile hypertrophic pyloric stenosis and congenital heart disease: an under recognized association. PMID- 9409171 TI - Ischemic stroke in sickle cell disease: a review. PMID- 9409172 TI - Help them to stand tall. PMID- 9409173 TI - A hypertensive man with abdominal pain. PMID- 9409174 TI - Report on the management of vesicoureteral reflux in children. PMID- 9409151 TI - Cell biology and molecular basis of denitrification. AB - Denitrification is a distinct means of energy conservation, making use of N oxides as terminal electron acceptors for cellular bioenergetics under anaerobic, microaerophilic, and occasionally aerobic conditions. The process is an essential branch of the global N cycle, reversing dinitrogen fixation, and is associated with chemolithotrophic, phototrophic, diazotrophic, or organotrophic metabolism but generally not with obligately anaerobic life. Discovered more than a century ago and believed to be exclusively a bacterial trait, denitrification has now been found in halophilic and hyperthermophilic archaea and in the mitochondria of fungi, raising evolutionarily intriguing vistas. Important advances in the biochemical characterization of denitrification and the underlying genetics have been achieved with Pseudomonas stutzeri, Pseudomonas aeruginosa, Paracoccus denitrificans, Ralstonia eutropha, and Rhodobacter sphaeroides. Pseudomonads represent one of the largest assemblies of the denitrifying bacteria within a single genus, favoring their use as model organisms. Around 50 genes are required within a single bacterium to encode the core structures of the denitrification apparatus. Much of the denitrification process of gram-negative bacteria has been found confined to the periplasm, whereas the topology and enzymology of the gram positive bacteria are less well established. The activation and enzymatic transformation of N oxides is based on the redox chemistry of Fe, Cu, and Mo. Biochemical breakthroughs have included the X-ray structures of the two types of respiratory nitrite reductases and the isolation of the novel enzymes nitric oxide reductase and nitrous oxide reductase, as well as their structural characterization by indirect spectroscopic means. This revealed unexpected relationships among denitrification enzymes and respiratory oxygen reductases. Denitrification is intimately related to fundamental cellular processes that include primary and secondary transport, protein translocation, cytochrome c biogenesis, anaerobic gene regulation, metalloprotein assembly, and the biosynthesis of the cofactors molybdopterin and heme D1. An important class of regulators for the anaerobic expression of the denitrification apparatus are transcription factors of the greater FNR family. Nitrate and nitric oxide, in addition to being respiratory substrates, have been identified as signaling molecules for the induction of distinct N oxide-metabolizing enzymes. PMID- 9409175 TI - Wegener's granulomatosis: histological documentation of common and uncommon manifestations in 216 patients. AB - BACKGROUND: Wegener's granulomatosis is the prototype of pulmonary angiitis and granulomatosis and a systemic vasculitic syndrome of unknown etiology. Wegener's granulomatosis can involve virtually any and often a multitude of organ-tissues. PATIENTS AND METHODS: This survey of 216 patients provides a histological documentation and pertinent literature review of both the common and uncommon, but with emphasis on the uncommon, manifestations of Wegener's granulomatosis. RESULTS: The common manifestations of the disease include the classic triad of upper airway, lung, and kidney, in 87%, 69%, and 48% of the patients, respectively. The less common manifestations involve the skin, central nervous system, eye and orbit, heart, breast, salivary gland, gastrointestinal tract, spleen, and male and female urogenital tracts; each of these accounts for less than 15% in all cases and below 5% for most of the patients. CONCLUSIONS: The manifestations of Wegener's granulomatosis in many of the uncommon anatomical sites of involvement may be distinctive or atypical and therefore, the histopathological diagnosis must be correlated with clinical and laboratory test findings. PMID- 9409176 TI - Morphological and functional changes of the arterial wall in subjects at risk of atherosclerosis and in patients with peripheral arterial occlusive disease. AB - BACKGROUND: The aim of this study was to determine the intima-media thickness (IMT) of the carotid arteries, which is regarded as the earliest morphological evidence of the atherosclerotic process in subjects with risk factors of atherosclerosis. Changes of blood flow in the branchial artery during reactive hyperemia were also investigated. PATIENTS AND METHODS: In 4 groups of subjects: smokers, diabetics, patients with peripheral arterial occlusive disease (PAOD) and controls, the above mentioned morphological and functional changes were studied by ultrasound. Each group comprised 18 subjects, aged 32 to 56 years. First, IMT was measured at three different sites of the carotid arteries. Then blood flow in the brachial artery was determined at rest and during reactive hyperemia (caused by handgrip test). RESULTS: In the control group the mean IMT (all segments) was 0.65 +/- 0.08 mm. IMT of smokers was similar (0.65 +/- 0.07 mm), while in diabetics a tendency to IMT increase was detected at all three measuring sites, and in the common carotid artery the IMT was statistically significantly thicker than in the controls (0.75 +/- 0.08 mm vs. 0.65 +/- 0.08, p < 0.05). PAOD patients showed a significant increase of IMT at all three carotid segments (mean 0.77 +/- 0.10 mm p < 0.05). In all groups IMT was related to the number of atherosclerotic plaques (r = 0.6, p < 0.001), and was also correlated with body mass index (BMI) (r = 0.32, p < 0.01) and fibrinogen level (r = 0.32, p < 0.05). All groups demonstrated a significant increase of blood flow in the branchial artery during hyperemia. In PAOD patients this increase was significantly smaller than in the controls and other groups (212 +/- 61% vs 275 +/- 60%, p < 0.05). CONCLUSIONS: The results of our study indicated that the most important factors determining increase of IMT are diabetes, fibrinogen level, BMI and the presence of clinically manifested atherosclerosis. Hyperemic blood flow, which is predominantly determinated by the vasodilation capacity of the resistance vessel, is reduced in patients with clinically manifested atherosclerosis and not in subjects with risk factors and without atherosclerotic manifestation. PMID- 9409177 TI - Cutaneous inflammation limited to the region of the ulcer in chronic venous insufficiency. AB - BACKGROUND: The role of inflammatory reactions in the pathogenesis of chronic venous insufficiency and the persistence of venous ulcerations is still not totally clear and remains a hotly debated topic. An investigation of the intensity and distribution of ICAM-1 expression and different inflammatory cells should help clarify whether inflammatory processes are limited locally to the area of the ulcer or if an upregulation can also be observed in clinically unaffected skin of CVI-III patients, as a sign of a primary inflammatory process. PATIENTS AND METHODS: We examined two skin areas in 10 patients with venous ulcerations. One area was at the border of the ulcer and another in clinically unaffected skin (distance from the ulcer: 12.6 +/- 5.1 cm). In addition skin specimens were obtained from the perimalleolar skin of 10 healthy controls. Our histological and immunohistochemical examinations were focused on inflammatory cells (B and T lymphocytes, macrophages, and mast cells) and on the adhesion molecule ICAM-1. RESULTS: A very strong expression of ICAM-1 could be seen at the border of the ulcer. This tissue also showed a dense infiltration, mainly by T lymphocytes and macrophages. In some cases the tissue was infiltrated by an increased number of mast cells. This is the typical picture of a chronic inflammatory reaction. Compared to healthy controls, the clinically unaffected skin of patients showed not an increased expression of ICAM-1 and only in some cases we could find a slight perivascular infiltrate of T lymphocytes. CONCLUSIONS: These data imply that the upregulation of endothelial adhesion molecules (ICAM-1) and dermal infiltration by T lymphocytes and macrophages in CVI-III patients is limited to the region of the ulcer, or at least to skin areas with a severe microangiopathy, and is part of a secondary elimination of necrotic issue (an 'injury and repair' process). These local chronic inflammatory reactions are certainly an important factor in the persistence and recurrence of venous ulcerations. PMID- 9409178 TI - Elastic compression stockings: durability of pressure in daily practice. AB - BACKGROUND: Elastic compression stockings are valuable tools in the treatment of many phlebological diseases. As to day no data are available about how long these elastic compression stockings continue to exert enough compression for a proper clinical function. PATIENTS AND METHODS: In this study we measured the pressure at the B level of compression class II and III elastic stockings directly after the patients started to use them, after 1 month, and after 3 months. In total 99 below knee stocking were measured with a TNO-tester, which has officially been accepted by the CEN for measurement of elastic compression stockings. RESULTS: Class II flat knitted stockings showed a mean pressure of 29.3 (SD 4.9) mmHg at the start of the study, declining to 27.6 (SD 5.2) mmHg, and 26.5 (SD 4.4) mmHg, at 1 and 3 months respectively. The round knitted class II had comparable results. In class III flat knitted stockings we measured 47.5 mmHg (SD 8.1) at the start, and 44.2 (SD 7.1) mmHg, and 41.3 (SD 6.7) mmHg, at 1 and 3 months respectively. Extrapolation of the results after 1 and 3 months demonstrate that the mean pressure of class II stocking is less than 25 mmHg after 4 to 5 months. Class III stockings were less than 35 mmHg close to 6 months, if evaluated in the same way. Individual regression analysis show that after 6 months 66% of the pressure class II stockings have a pressure of less than 25 mmHg and 45% of the pressure class III less than 35 mmHg. When considering a 10% safety margin the figures are respectively 84% and 63%. CONCLUSIONS: From the results of this study it can be concluded that, especially for the most frequently used compression class II stockings, three new stockings each year are necessary to ensure an effective function of the stocking during the time they are being used by the patient. For compression class III flat knitted stockings two pairs can be considered as sufficient. PMID- 9409179 TI - Circadian pattern of post-surgical fatal pulmonary embolism. AB - BACKGROUND: The circadian distribution of fatal pulmonary thromboembolism in general surgical patients is unknown. PATIENTS AND METHODS: One hundred consecutive cases of pulmonary embolism, with reliable clinical notes and data, were studied (67 men and 33 women; mean age 71 years). Only post-surgical cases were considered in this analysis. Patients had undergone elective (78%) or emergency abdominal surgery (22%). Correct prophylaxis (according to the Windsor Consensus Statement) had been used in 12%. Cases were grouped according to the time of onset of signs and symptoms related to pulmonary embolism at one hour intervals. RESULTS: The maximum incidence of fatal pulmonary embolism was between 7.00 a. m. and 1.00 p. m. with the highest peaks at 9.00 and 11.00 a. m. 9% of deaths) (P < 0.02). When results from this study were compared to a previous study no significant difference was observed between the distribution profile of cases from general medical wards and surgical wards. CONCLUSION: It appears that in surgical patients there is a circadian pattern in pulmonary embolism as already documented in medical patients. PMID- 9409180 TI - Development and validation of a disease-specific questionnaire on the quality of life of patients with chronic venous insufficiency. AB - BACKGROUND: Recording of Quality of Life (QoL) is taking on increasing importance in medicine. Although QoL cannot be measured directly, numerous methods studies have demonstrated that important areas of QoL can be addressed by validated questionnaires. The present study was designed to examine a new disease-specific, German-language questionnaire, the Freiburger Questionnaire of QoL in venous diseases (FLQA), with respect to reliability, validity and patient acceptance among patients with chronic venous insufficiency (CVI). PATIENTS AND METHODS: The FLQA consists of 83 items and differentiates between limitations in QoL in 7 scales: physical complaints, everyday life, social life, emotional status, therapy, satisfaction, occupation. Data were collected from 246 patients with CVI stage I (n = 107), II (n = 75) and III (venous ulcer, n = 64) in the Widmer classification. RESULTS: In the psychometric test, the majority of scales showed little or no top or bottom effect. The internal consistency of the scales was generally high (Cronbach's Alpha in all scales more than alpha = .70 and in 4 scales above alpha = .90). The convergent validity with the Nottingham Health Profile and the ALLTAG was high in several scales. The FLQA showed good discriminant validity and significantly differentiated between the clinical CVI stages in several areas of QoL. The change sensitivity during the course of therapy was also good. In a feasibility test, the acceptance values among patients were high with respect to ease of understanding and description of the QoL areas relevant to them. The mean time to fill out the questionnaire was 21 +/ 6 minutes. CONCLUSION: Overall, the FLQA appears suitable to reliably record characteristics of QoL in patients with CVI in both course and cross-sectional studies. PMID- 9409181 TI - Successful treatment of an extended leg ulcer in systemic sclerosis. AB - We report the successful surgical treatment of a large and painful leg ulcer associated with systemic sclerosis (scleroderma). In addition, there was a long occlusion of the superficial femoral artery, and ankle systolic blood pressure was 80 mmHg (ankle-brachial-index 0.65). All conservative treatments including systemic antibiotics, nifedipine, intravenous iloprost, intravenous penicilline G and hyperbaric oxygen failed. Pain was intolerable and below-knee amputation was considered. In a first attempt to save the limb, the patient underwent femoropopliteal bypass surgery. Despite a successful outcome of the bypass operation and normalization of the ankle blood pressure, the large wound remained recalcitrant and extremely painful. A second attempt to save the limb consisted of complete debridement of all sclerotic tissue down to the fascia and split skin grafting. The graft took in over 90% of the surface and the remaining wound healed spontaneously. Large leg ulcers in systemic sclerosis can become limb threatening. Radical debridement combined with a split skin graft seems to be a promising way to avoid amputation. PMID- 9409182 TI - An unusual cause of upper gastrointestinal haemorrhage--perforation of a vena cava filter into the duodenum. AB - Penetration of vena cava inferior filter spikes through the vessel wall is a frequent but only rarely symptomatic complication of these devices. We report a case of upper gastrointestinal haemorrhage 11 year after implantation of a Mobin Uddin filter. At gastroduodenoscopy a filter spike protruding from the duodenal mucosa was identified as the cause of the haemorrhage. After removal of the filter the patient made an uneventful recovery. Indications for vena cava filter placement are discussed in the light of their early and late complications. Surgical intervention is strongly recommended for symptomatic perforations. PMID- 9409183 TI - Pseudocoarctation of the abdominal aorta. AB - The case of an asymptomatic pseudocoarctation of the abdominal aorta is reported. Pronounced kinking of the abdominal aorta was seen on angiograms, without stenosis or collateral circulation. Angiography revealed normal renal arteries originating from the malformated aortal segment. The possible etiology and the need for the therapeutical intervention are discussed. PMID- 9409185 TI - Global AIDS surveillance--part I. PMID- 9409184 TI - Is intermittent claudication in patients with peripheral arterial occlusive disease relieved under hyperbaric conditions? PMID- 9409186 TI - The current global situation of the HIV/AIDS pandemic. PMID- 9409187 TI - WHO scientific group meeting on cardiovascular disease and steroid hormone contraceptives. PMID- 9409188 TI - Adjuvant therapy in cervical cancer patients with high risk factors. AB - Neoadjuvant and adjuvant chemotherapies are used adjunctively with surgery or radiation and are among the treatment options that are now employed for reducing treatment failure in early-stage cervical cancers with high-risk prognostic factors. Adjuvant therapies have been reported to significantly improve survival than would otherwise be possible with surgery or radiotherapy alone. However, for advanced cervical cancers, sequential or concurrent chemo-radiotherapy does not appear to significantly increase survival. The combination of radiotherapy with IFN-a2a and RA in the treatment of patients with locally advanced cervical cancer showed high response rates, however this should be confirmed in larger studies. Recent reports show that postoperative adjuvant radiotherapy has no benefit in survival, but that postoperative adjuvant chemotherapy has improved survival. Toxicities and the optimum number of cycles of neoadjuvant and adjuvant chemotherapy, as well as biologic therapy, will follow along with individualized treatment based on high-risk prognostic factors. Although more comprehensive studies and longer follow up will be required for complete evaluation of these adjuvant therapies, preliminary results are promising. PMID- 9409189 TI - Role of nitric oxide in penile erection. AB - The present study was undertaken to investigate the role of nitric oxide (NO) in erectile physiology by correlating its action with the existence and activity of nitric oxide synthase (NOS), which produces NO. We applied Western blot analysis in both human and rat penile tissue. In the rat, reduced nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase staining and spectrophotometric assay were also performed, in addition to in vivo electroerection study with pharmacological manipulation. Western blot analysis identified a protein of 155 KDa identical to the neural form of NOS in the human and rat penis. The NOS blot densities in the two species were similar, and both were lower than that in the rat cerebellum. Histochemical staining localized NOS to neurons innervating the corpora cavernosa, including the pelvic plexus, the cavernosal nerves and their terminal fibers within the corporeal erectile tissue, and dorsal penile nerves. NOS activity was also found in the cerebellum, urethra, penis, and urinary bladder, in decreasing order of intensity. Intracavernous injections of NOS inhibitor (L-NOARG or L-NAME in concentrations from 10(-6) M to 10(-3) M suppressed electrostimulation-induced erection in a concentration-dependent manner. Subsequent intracavernous injection of L-Arginine (10(-2) M) partially restored the erection. The neural form of constitutive NOS in the corpora cavernosa synthesizes NO, which mediates penile erection. Determination of cavernosal NOS expression or activity may permit characterization of certain pathological conditions that cause impotence. PMID- 9409190 TI - Eight-year experience of malignant lymphoma--survival and prognostic factors. AB - Several reports have suggested a geographic difference in the histopathologic characteristics and prognosis of malignant lymphoma around the world. We tried to evaluate the clinical and histopathologic characteristics, therapeutic outcomes, and prognostic features of malignant lymphoma, particularly in Korean patients. Three hundred and seventy-six adult patients with the initial histopathologic diagnosis of malignant lymphoma of Yonsei University College of Medicine over an 8-year period were analyzed, retrospectively, with the following results: 1) There were 47 cases of Hodgkin's disease (HD) (12.5%) and 329 of non-Hodgkin's lymphoma (NHL) (87.5%) with a 1:7 ratio. The most common histopathologic subtype of HD was mixed cellularity (44.7%), and that of NHL was intermediate grade (70.8%), especially diffuse large-cell type (44.1%), whereas follicular type was less common. In regard to the incidence of extranodal presentation, it is rare in HD (4.2%), but occurs in 49.8% of patients with NHL. 2) The complete remission (CR) rate was 91.5% in HD and 63.6% in NHL, and the 5-year and 7-year disease free survival rates were 71.3% and 57.0% in HD; 67.0% and 49.6% in NHL. The 5 year and 8-year overall survival rates were 90.7% and 68.0% in HD; 65.2% and 60.2% in NHL. 3) By multivariate analysis, we found that age, performance status, histopathologic grade, stage, serum lactate dehydrogenase (LDH) and beta 2 microglobulin were the useful prognostic factors in predicting survival in NHL, while no definite prognostic factors were found in HD. Also, in NHL patients less than 60 years old, stage, serum LDH, and histopathologic grade were closely associated with their therapeutic outcomes. In conclusion, the characteristics of malignant lymphoma in our hospital differ from those in Western countries with respect to the clinical, histopathologic and immunophenotypic patterns, but the prognostic factors and overall therapeutic outcomes were quite comparable to those of other reports from Western countries. PMID- 9409191 TI - Granzyme B immunoreactivity in T/natural killer cell lymphomas. AB - Granzyme B is one of the serine proteases expressed in natural killer (NK) cells and activated cytotoxic T-lymphocytes. To evaluate the usefulness of granzyme B immunoreactivity in the diagnosis of T/NK-cell lymphoma, we studied 69 cases of lymphomas occurring in the upper aerodigestive tract by paraffin-section immunohistochemistry using a commercially available monoclonal antibody to granzyme B (GrB-7). All 19 cases of T/NK-cell lymphomas defined by the expression of CD56 (NHK-1) and one or both T-cell markers (polyclonal CD3 and CD45RO) expressed granular cytoplasmic granzyme B immunoreactivity. Two out of 9 cases of T-cell lymphomas showing no CD56 expression demonstrated strong granzyme B immunoreactivity. No tumor cells among 39 cases of B-cell diffuse large cell lymphomas and 2 cases of null cell diffuse large cell lymphomas were immunoreactive for granzyme B, however a few scattered granzyme B-positive reactive small lymphoid cells were consistently observed. Based on its sensitivity in this study as well as its reactivity in routinely processed tissue sections, even without heat-induced epitope retrieval technique, monoclonal antibody to granzyme B (GrB-7) can be applied as a useful marker in the diagnosis of T/NK-cell lymphomas in conjunction with CD56. PMID- 9409192 TI - Arthroscopic transglenoid Bankart suture repair with modifications of Caspari's technique. AB - Recent advances in arthroscopic surgery have given new options for dealing with anterior glenohumeral instability with less morbidity than the open procedure. The early literature discussing arthroscopic Bankart suture repair is favorable, however limited long-term follow-up studies have yet to prove its success. This study reports our experience with arthroscopic transglenoid Bankart suture repair with a minimum 2-year follow-up. Arthroscopic transglenoid Bankart repair was performed in 23 cases of anterior shoulder instability with some modifications of Caspari's suture technique. First, the suture material was #1 PDS and the number of sutures was 4 or 5. Second, the insertion site of the guide pin was moved to 1 o'clock on the right shoulder and to 11 o'clock on the left shoulder to prevent slippage of the guide pin. With this technique, we obtained 87% satisfactory results analyzed by the Rowe functional grading system. There was no suprascapular or axillary nerve injury. Two patients had redislocations and one patient had recurrent subluxation. Five patients complained of suture-knot irritation problems. PMID- 9409193 TI - Cyclin E expression in benign and malignant epithelial neoplasms of the gallbladder. AB - Cyclins are the regulatory subunits of cyclin-dependent kinase and play an important role in cell proliferation. Many tumors, such as colon, breast and gastric carcinomas are known to be involved in the deregulation or amplification of cyclins, especially cyclin E, which involves the restriction point of G1-S transition. We investigated the expression of cyclin E in benign and malignant epithelial tumors of the gallbladder and compared the results with the activity of cell proliferation by the Ki67 antigen using immunohistochemical staining. Cyclin E was expressed in the adenocarcinoma tissue in 33.3% of patients (4 out of 12 cases), whereas only one out of 8 cases of adenoma expressed cyclin E (12.5%). There was a correlation between cyclin E expression and the Ki67 labeling index. These results suggest that the high expression of cyclin E in adenocarcinoma of the gallbladder is related to a high rate of cell proliferation. PMID- 9409194 TI - Long-term results of cyclosporine-induced remission of relapsing nephrotic syndrome in children. AB - Twenty-nine children with nephrotic syndrome were treated with cyclosporine (CsA), 100 mg/m2/day for 6 months and prednisone, 2 mg/kg every other day for 1 month and then subsequently 1 mg/kg every other day for 5 months. A renal biopsy had shown minimal change disease (MCD) in 18 children, focal segmental glomerulosclerosis (FSGS) in 3 children, membranous glomerulonephritis (MGN) in 4 children, membranoproliferative glomerulonephritis (MPGN) in 2 children, and IgA nephropathy in 2 children. All MCD patients went into complete remission during therapy. Five out of 11 steroid-sensitive patients (45.5%) remained in complete remission, while the remaining 6 (54.5%) had 2 to 3 relapses, 19 to 47 months after CsA discontinuation. Two out of 7 steroid-resistant patients (28.6%) were still in complete remission and 5 (71.4%) had 1 to 6 relapses 25 to 49 months after CsA withdrawal. The mean number of relapses in the steroid-sensitive group before and after CsA treatment decreased more (8.5 vs 1.4) than in the steroid resistant group (8.1 vs 2.4) (p < 0.05). At the most recent examination, 1 of 3 FSGS patients achieved complete remission and 2 had a partial response. Three of 4 MGN patients were in complete remission and 1 was in partial remission. One of 2 MPGN patients achieved complete remission and 1 showed partial remission. Two patients with IgA nephropathy were in partial remission. We compared MCD patients in sustained remission and relapse; the mean CD4/CD8 ratio decreased from 1.5 to 0.9 in the remission group, in comparison with no change in the relapsed group (p < 0.05). The posttreatment renal biopsy showed lesions of nephrotoxicity in 3 of 18 children with MCD whose renal function did not alter after CsA treatment. We concluded: 1) A 6-month treatment of CsA, in combination with a low-dose alternate-day steroid, proved to be effective in maintaining the remission of steroid-sensitive and steroid-resistant MCD patients. 2) The CD4/CD8 ratio can be used as a index to predict remission or relapse after CsA therapy. PMID- 9409195 TI - Primary hypertrophic osteoarthropathy accompanied by Crohn's disease: a case report. AB - Primary hypertrophic osteoarthropathy is a rare hereditary disease without evidence of underlying diseases. We report a very unusual case of primary HOA accompanied by Crohn's disease with the primary HOA mimicking secondary HOA, which is a rare manifestation of Crohn's disease. We also review the literature to find the correlation, if any, between the two. PMID- 9409196 TI - Pulmonary asbestosis: radiologic-pathologic brief report. AB - Pulmonary asbestosis is defined as bilateral diffuse interstitial fibrosis of the lungs caused by exposure to asbestos. Many occupations are at risk for asbestos exposure, particularly in the mining, milling, manufacturing, construction, shipbuilding, and automotive industries. Therefore, the prevalence of asbestosis should be fairly widespread. The diagnosis of asbestosis can be made on either clinical or pathological grounds. We recently encountered one case of asbestosis which was confirmed histologically. On HRCT, there was ground-glass opacity with irregular linear shadows, subpleural curvilinear lines and parenchymal bands. Neither plaque nor calcification were noted. The histologic findings observed on open-lung biopsy specimen were well in accord with those in HRCT. Many asbestos coated bodies were present along with black dust. PMID- 9409197 TI - Isoprostanes: potential markers of oxidant stress in atherothrombotic disease. AB - Isoprostanes are emerging as a new class of biologically active products of arachidonic acid metabolism of potential relevance to human vascular disease. Their formation in vivo seems to reflect primarily, if not exclusively, a nonenzymatic process of lipid peroxidation. Enhanced urinary excretion of 8-iso PGF2 alpha has been described in association with cardiac reperfusion injury and with cardiovascular risk factors, including cigarette smoking, diabetes mellitus, and hypercholesterolemia. Besides providing a likely noninvasive index of lipid peroxidation in these settings, measurements of specific F2 isoprostanes in urine may provide a sensitive biochemical end point for dose-finding studies of natural and synthetic inhibitors of lipid peroxidation. Although the biological effects of 8-iso-PGF2 alpha in vitro suggest that it and other isoeicosanoids may modulate the functional consequences of lipid peroxidation, evidence that this is likely in vivo remains inadequate at this time. PMID- 9409198 TI - Effect of phosphorothioated oligonucleotides on neointima formation in the rat carotid artery. Dissecting the mechanism of action. AB - Several studies have shown that single-dose administration of agents that inhibit medial cell replication, such as antisense oligonucleotides to cell replication genes, can inhibit neointima formation after arterial injury. However, the precise mechanism of action of these agents is unknown. We analyzed the effect of phosphorothioated oligonucleotides delivered periadventitially on the response to injury in the balloon-injured rat carotid artery. Antisense oligonucleotides to c myc suppressed medial replication 2 days after injury, but this effect was not present at 4 or 14 days. Endothelial cell proliferation was not affected by antisense oligonucleotides. There was, however, a significant suppression of intimal area and intima/media ratio at 14 days and an increase in lumen area in the antisense-treated group. Indeed, an increase in the number of medial cells at 14 days in the antisense group indicated that most of the effect of the agent was due to the suppression that most of the effect of the agent was due to the suppression of cell migration. No effect was noted on expression of two genes, osteopontin and tropoelastin, used as markers of modulation of smooth muscle cells to a "neonatal" phenotype at 4 days after injury. Because no effect on cell proliferation could be demonstrated after 2 days, our data indicate that an early effect of the antisense agent mediates its longer-term effects. We suggest that this effect may be due to the suppression of migration of medial smooth muscle cells rather than the suppression of medial or intimal cell proliferation. PMID- 9409199 TI - Atherosclerosis, vascular remodeling, and impairment of endothelium-dependent relaxation in genetically altered hyperlipidemic mice. AB - We examined the vascular structure and endothelium-dependent relaxation in two genetic models of hypercholesterolemia: apolipoprotein E (apoE)-knockout mice and combined apoE/LDL receptor-double-knockout mice. Intimal area was increased markedly in proximal segments of thoracic aortas from apoE/LDL receptor-knockout mice [0.13 +/- 0.03 (mean +/- SE) mm2] compared with normal (C57BL/6J) mice (0.002 +/- 0.002 mm2, P < .05). Despite intimal thickening, the vascular lumen was not smaller in the aortas of apoE/LDL receptor-knockout mice (0.52 +/- 0.03 mm2) than in normal mice (0.50 +/- 0.03 mm2). In apoE-deficient mice, intimal thickening was minimal or absent, even though the concentration of plasma cholesterol was only modestly less than that in the double-knockout mouse (14.9 +/- 1.1 vs 18.0 +/- 1.2 mmol/L, respectively, P < .05). Relaxation of the aorta was examined in vitro in vascular rings precontracted with U46619. In normal mice, acetylcholine produced relaxation, which was markedly attenuated by the nitric oxide synthase inhibitor NG-nitro-L-arginine (100 microM). Relaxation to acetylcholine and the calcium ionophore A23187 was normal in apoE-deficient mice (in which lesions were minimal) but greatly impaired in the proximal segments of thoracic aortas of apoE/LDL receptor-deficient mice, which contained atherosclerotic lesions. Vasorelaxation to nitroprusside was similar in normal and apoE-knockout mice, with modest but statistically significant impairment in atherosclerotic segments of apoE/LDL receptor-knockout mice. In distal segments of the thoracic aorta of apoE/LDL receptor-deficient mice, atherosclerotic lesions were minimal or absent, and the endothelium-dependent relaxation to acetylcholine and calcium ionophore was normal. Thus, in apoE/LDL receptor knockout mice (a genetic model of hyperlipidemia), there is vascular remodeling with preservation of the aortic lumen despite marked intimal thickening, with impairment of endothelium-dependent relaxation to receptor- and nonreceptor mediated agonists. Atherosclerosis may be accelerated in the apoE/LDL receptor double-knockout mouse compared with the apoE-knockout strain alone. We speculate that other factors, such as the absence of LDL receptors, may contribute to the differences in the extent of atherosclerosis in these two models of hyperlipidemia. PMID- 9409200 TI - CLA-1 is an 85-kD plasma membrane glycoprotein that acts as a high-affinity receptor for both native (HDL, LDL, and VLDL) and modified (OxLDL and AcLDL) lipoproteins. AB - Lipoprotein metabolism is regulated by the functional interplay between lipoprotein components and the receptors and enzymes with which they interact. Recent evidence indicates that the structurally related glycoproteins CD36 and SR BI act as cell surface receptors for some lipoproteins. Thus, CD36 has been reported to bind oxidized LDL (OxLDL) and acetylated LDL (AcLDL), while SR-BI also binds native LDL and HDL. The cDNA of human CLA-1 predicts a protein 509 amino acids long that displays a 30% and an 80% amino acid identity with CD36 and mouse or hamster SR-BI, respectively. In this report, we describe the structural characterization of CLA-1 as an 85-kD plasma membrane protein enriched in N linked carbohydrates. The expression of CLA-1 on mammalian and insect cells has been used to demonstrate that CLA-1 is a high-affinity specific receptor for the lipoproteins HDL, LDL, VLDL, OxLDL, and AcLDL. Northern blot analysis of the tissue distribution of CLA-1 in humans indicated that its expression is mostly restricted to tissues performing very active cholesterol metabolism (liver and steroidogenic tissues). This finding, in the context of the capability of this receptor to bind to both native and modified lipoproteins, strongly suggests that the CLA-1 receptor contributes to lipid metabolism and atherogenesis. PMID- 9409201 TI - High-density lipoprotein-binding protein (HBP)/vigilin is expressed in human atherosclerotic lesions and colocalizes with apolipoprotein E. AB - Accumulation of cholesteryl esters within cells of the arterial intima is a hallmark of atherosclerosis. A small number of proteins have been shown in vitro to be upregulated by cellular cholesterol loading, including apolipoprotein E (apoE) and the recently cloned HDL-binding protein (HBP), but only apoE has been shown to be upregulated in cholesterol-loaded cells in atherosclerosis. To determine whether HBP (also called vigilin) might be expressed in human atherosclerosis, immunohistochemistry and in situ hybridization were performed on coronary arteries of 18 patients. HBP/vigilin was detected on all endothelial cells. HBP/vigilin mRNA and protein also were detected on a subset of macrophages and occasionally on smooth muscle cells (SMC) in atherosclerotic plaques but were not detected on these cell types in nondiseased coronary intima. The majority of HBP/vigilin-expressing macrophages were foam cells, but HBP/vigilin expression also was detected rarely in nonfoam cell macrophages. Foam cell macrophage HBP/vigilin expression was present in 100% of atherosclerotic quadrants, and nonfoam cell macrophage HBP/vigilin expression was present in 6% of atherosclerotic quadrants. HBP/vigilin-expressing human plaque cells also expressed apoE. However, HBP/vigilin was detected in cardiac myocyte foam cells of an apoE-deficient mouse, demonstrating that HBP/vigilin expression can occur independently of apoE. These results suggest that in vivo HBP/vigilin expression is upregulated by intracellular cholesterol loading but also that other factors present in atherosclerotic plaques may upregulate HBP/vigilin. Although the exact function of HBP/vigilin is unknown, its expression in plaque macrophages suggests a role for this molecule in atherogenesis. PMID- 9409202 TI - Calreticulin, a potential vascular regulatory protein, reduces intimal hyperplasia after arterial injury. AB - Both thrombotic and inflammatory responses to arterial injury have been implicated in atherosclerotic plaque growth. Calreticulin is a ubiquitous calcium binding protein with antithrombotic activity and, in addition, is associated with leukocyte activation. We are investigating calreticulin as a potential vascular regulatory protein. The development of intimal hyperplasia was studied at sites of balloon injury in iliofemoral arteries from 91 rats. Calreticulin was infused directly into the artery immediately before balloon injury, and plaque growth was then assessed at 4 weeks' follow-up. Parallel studies of the effects of each calreticulin domain as well as a related calcium-binding protein, calsequestrin, were examined. The effects of calreticulin on platelet activation, clot formation, and mononuclear cell migration were also studied. When infused before balloon injury in rat iliofemoral arteries, calreticulin, or its high-capacity Ca(2+)-binding C domain, significantly reduces plaque development, whereas calsequestrin, a related calcium-binding protein that lacks the multifunctional nature of calreticulin, does not decrease plaque area (saline: 0.037 +/- 0.007 mm2, calsequestrin: 0.042 +/- 0.021 mm2, calreticulin: 0.003 +/- 0.002 mm2, n = 46, P < .04). The N domain and more specifically the P domain, a low-capacity, high-affinity calcium-binding domain in calreticulin, do not reduce intimal hyperplasia (N + P domain: 0.038 +/- 0.012 mm2, C domain: 0.003 +/- 0.002 mm2, n = 45 rats, P < .0001). Calreticulin reduces macrophage and T cell staining in the arterial wall after injury but has no direct effect on monocyte migration in vitro (percent medial area staining positive for macrophage 24 hours after injury (N + P: 4.06 +/- 1.42, calreticulin: 0.273 +/- 0.02; n = 26, P < .009). Calreticulin does, however, reduce platelet-dependent whole blood clotting time, in vitro (baseline: 78.23 +/- 2.04 seconds, calreticulin: 113.5 +/- 1.95 seconds; n = 5, P < .002). We conclude that calreticulin significantly reduces intimal hyperplasia after arterial injury, potentially acting as a vascular regulatory protein. PMID- 9409203 TI - Scavenger receptors are present on rabbit aortic endothelial cells in vivo. AB - Endothelial cells metabolize modified LDL, but attempts to detect scavenger receptors in this cell type in vitro have been unsuccessful. To determine whether scavenger receptors are present on endothelial cells in vivo, species-specific reagents were developed to detect rabbit scavenger receptor protein. Antiserum against the rabbit scavenger receptor was generated with the use of synthetic peptides of two distinct regions: residues 3 to 21 in the cytoplasmic tail and residues 282 to 304 in the collagen-like region. Reactivity of antiserum against the synthetic peptides was confirmed with an enzyme-linked immunosorbent assay. Positive reactivity was also observed against fragments of scavenger receptor protein expressed in bacteria. Antiserum to both regions reacted with liver membrane proteins of sizes consistent with the scavenger receptor, as confirmed by Western blotting under reduced and nonreduced conditions. Immunocytochemical examination of rabbit aortic tissue by use of antiserum to both regions of scavenger receptor protein produced striking and identical patterns of staining of aortic endothelium. Immunostaining was abolished for both antisera by preadsorption with the specific peptide region used as immunogen. In contrast, incubation of scavenger receptor antiserum with a peptide of a region of the rabbit LDL receptor failed to influence immunoreactivity against endothelium. These data demonstrate the presence of scavenger receptors in rabbit endothelium in vivo, which may have fundamental implications for lipoprotein metabolism by the arterial wall. PMID- 9409204 TI - beta-VLDL hypercholesterolemia relative to LDL hypercholesterolemia is associated with higher levels of oxidized lipoproteins and a more rapid progression of coronary atherosclerosis in rabbits. AB - The accumulation of the oxidized apolipoprotein, apoB-100, containing lipoproteins in the arterial wall and the progression of coronary atherosclerotic lesions in rabbits with beta-VLDL and LDL hypercholesterolemia was compared. In New Zealand White (NZW) rabbits on a 0.125% cholesterol diet, LDL cholesterol levels increased from 14 +/- 1 mg/dL (mean +/- SEM; n = 9) to 170 +/- 34 mg/dL (n = 10, P = .0002). On 0.5% cholesterol, LDL cholesterol levels were similar, but beta-VLDL cholesterol levels increased from 60 +/- 4 mg/dL (n = 10) to 550 +/- 75 mg/dL (n = 8; P < .0001). In Watanabe heritable hyperlipidemic (WHHL) rabbits, LDL cholesterol levels were 2.3-fold higher (n = 13; P < .0001) than in NZW rabbits on 0.5% cholesterol, whereas their beta-VLDL cholesterol levels were 3.7 fold lower (P < .0001), resulting in similar total cholesterol levels. At 2 months, mean intimal areas of lesions in the coronary arteries of NZW rabbits on 0.125% cholesterol were 0.13 +/- 0.045 mm2 (n = 4; mean +/- SEM) and were 5.8 fold, (n = 4; P = .016) and 2.0-fold (n = 6; P = NS versus 0.125% cholesterol and P = .014 versus 0.5% cholesterol) higher in NZW rabbits on 0.5% cholesterol and in WHHL rabbits, respectively. At 5 months, mean intimal areas were 0.47 +/- 0.088 mm2 (n = 6) in NZW rabbits on 0.125% cholesterol and were 4.5-fold (n = 4; P = .0001) and 2.0-fold (n = 7; P = .012 and P = .0019) higher in rabbits on 0.5% cholesterol and in WHHL rabbits, respectively. Levels of oxidized apoB-100 containing lipoproteins (both beta-VLDL and LDL) in the lesions correlated with mean intimal area (r = .88; n = 31; P < .0001) of those lesions and with the plasma levels of total beta-VLDL/LDL (r = .72; P < .0001). Levels of oxidized apoB-100 containing lipoproteins in the arterial wall correlate with progression of hypercholesterolemia-induced coronary atherosclerotic lesions. Plasma levels of beta-VLDL relative to similar increases in LDL result in a more pronounced accumulation of oxidized apoB-100 containing lipoproteins in the arterial wall and in the plasma and a more rapid progression of coronary atherosclerosis. PMID- 9409205 TI - Apoptosis after stent implantation compared with balloon angioplasty in rabbits. Role of macrophages. AB - Both cell proliferation and apoptosis (programmed cell death) are supposed to play a role in restenosis after angioplasty. We studied these processes in smooth muscle cells (SMCs) and macrophages 1, 4, and 12 weeks after balloon angioplasty or Palmaz-Schatz stent implantation in rabbit iliac arteries. Proliferating cells were visualized by immunostaining with antibodies directed against proliferating cell nuclear antigen. Apoptotic cells were detected using the TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick and labeling) technique, propidium iodide staining, and transmission electron microscopy. At all time points, the neointimal cross-sectional area of the arteries was twofold to fourfold greater after stent implantation than after balloon angioplasty. The total number of neointimal cells was similar 1 and 12 weeks after both interventions. The neointimal cell density, however, decreased by 58% between the 1st and the 12th week after stent implantation compared with a 20% decrease after balloon angioplasty (P < .01). Stent implantation induced more cell proliferation but also more apoptosis in the media than balloon angioplasty after 1 and 4 weeks. In addition, stent implantation caused more macrophage accumulation and apoptosis in the neointima, but cell proliferation rates did not differ significantly in comparison with balloon angioplasty. The higher rate of apoptosis in the neointima 1 week after stent implantation compared with balloon angioplasty is due to an increased rate of SMC and macrophage death. Macrophage accumulation and apoptosis in the early phase after stent implantation appear to play a role in extracellular matrix secretion, which increases neointima formation after 4 and 12 weeks compared with balloon angioplasty in this model. PMID- 9409206 TI - Role of activin-A and follistatin in foam cell formation of THP-1 macrophages. AB - Macrophage (M phi) foam cell formation is a characteristic event that occurs in the early stage of atherosclerosis. To examine the roles of activin-A, a member of the transforming growth factor-beta superfamily, and follistatin, the binding protein for activin-A, in M phi function, we investigated their effects on foam cell formation of THP-1 M phi s. When THP-1 M phi s were treated with activin-A (5 nmol/L), foam cell formation and cellular cholesteryl ester accumulation were decreased. This downregulation was paralleled by a reduction in cell association and degradation of acetylated LDL. The inhibitory effect of activin-A on cell association and degradation was dose dependent, and the effect was blocked by concomitant addition of follistatin. Activin-A (5 nmol/L) also decreased the Bmax for acetylated LDL and scavenger receptor mRNA expression. Follistatin showed an effect opposite to that of activin-A and promoted M phi foam cell formation and cellular cholesteryl ester accumulation. It increased binding, cell association, and degradation of acetylated LDL and upregulated scavenger receptor mRNA expression. Because follistatin is the binding protein for activin-A, follistatin's effect is considered to be mediated by blocking the inhibitory effect of intrinsic activin-A. These results indicate that activin-A inhibits and follistatin promotes M phi foam cell formation by regulating scavenger receptor mRNA expression. We conclude that activin-A and follistatin play important roles in the process of atherosclerosis by regulating M phi foam cell formation. PMID- 9409207 TI - Quantitation of platelet-derived growth factor receptors in human arterial smooth muscle cells in vitro. AB - Platelet-derived growth factor (PDGF) is suggested to play an important role in the development of atherosclerosis as a migratory and mitogenic stimulus to arterial smooth muscle cells (ASMCs). Stimulated and unstimulated ASMCs were studied with respect to PDGF receptor (PDGF-R) mRNA and protein expression. Quantitative RT-PCR was developed for simultaneous evaluation of both PDGF-R alpha and -R beta mRNA expression and a quantitative ELISA for estimation of corresponding PDGF-R subunits. On the mRNA level, the overall PDGF-R beta expression was approximately 100 times lower than that of PDGF-R alpha. Furthermore, although PDGF-R alpha mRNA levels were high irrespective of hASMC phenotype, PDGF-R beta mRNA was influenced by serum stimulation with lower copy numbers in proliferating and confluent cells compared with quiescent cells. On the protein level, quiescent hASMCs expressed 10 times more PDGF-R beta than PDGF R alpha. Serum stimulation decreased cell surface PDGF-Rs, with most prominent loss of PDGF-R alpha (ELISA and immunohistochemistry). Our results suggest a differential regulatory pattern for PDGF-R alpha and -R beta and are compatible with the usage of alternative promoters for regulation of -R alpha expression. Further, it seems that the number of available receptor subunits is not the only determinant of variations in cell stimulation with different PDGF isoforms. PMID- 9409208 TI - Expression and function of recombinant endothelial NO synthase in coronary artery smooth muscle cells. AB - Smooth muscle cells (SMCs) play a key role in the pathogenesis of vascular diseases. The objectives of this study were to determine whether transfer of recombinant endothelial nitric oxide synthase (eNOS) gene to porcine coronary artery smooth muscle cell (CSMCs) would result in expression of a functional enzyme and to assess the effect of expression of eNOS on cell proliferation. CSMCs were transduced in vitro with adenoviral vectors encoding cDNA for eNOS (AdeNOS) and beta-galactosidase (Ad beta Gal). In contrast to Ad beta Gal- or sham-transduced cells, CSMCs transduced with AdeNOS stained positive with the NADPH-diaphorase stain, acquired calcium-dependent NOS activity (measured by the conversion of [3H]L-arginine to [3H]L-citrulline), had increasing cyclic 3',5' cGMP levels with increasing concentrations of the vector, and produced increased amounts of nitrite. cGMP production by AdeNOS-transduced cells was augmented by increasing intracellular levels of the eNOS cofactor tetrahydrobiopterin. CSMCs transduced with AdeNOS showed diminished serum-stimulated DNA synthesis as measured by thymidine uptake. Cell proliferation was diminished in AdeNOS transduced CSMCs as assessed by cell counts 3 and 6 days after serum stimulation of quiescent CSMCs. The present study demonstrates that adenovirus-mediated gene transfer of eNOS to CSMCs results in the expression of a functional enzyme whose activity can be augmented by increasing intracellular levels of tetrahydrobiopterin. Expression of recombinant eNOS in CSMCs results in inhibition of serum-stimulated DNA synthesis and cell proliferation. These findings imply that eNOS gene transfer to SMCs may be a unique mode of increasing local NO production in the arterial wall. PMID- 9409209 TI - Does insulin resistance unite the separate components of the insulin resistance syndrome? Evidence from the Miami Community Health Study. AB - A number of coronary heart disease risk factors have been identified that often cluster together to increase the risk of macrovascular disease. This cluster is referred to as the insulin resistance syndrome, and the risk factors commonly include dyslipidemia, elevated blood pressure, an android pattern of body fat distribution, and glucose intolerance. Whether hyperinsulinemia or insulin resistance per se provides a common pathway for these metabolic abnormalities is unclear. The authors studied 50 nondiabetic persons who had completed a euglycemic hyperinsulinemic clamp protocol in addition to a 75-g oral glucose tolerance test and other measures of the coronary risk profile. Using principal component analysis, we reduced nine coronary risk factors to two uncorrelated factors that explained 54.5% of the variance. Factor 1 consisted of positive loadings for uric acid, systolic and diastolic blood pressure, triglyceride concentration, and waist girth and negative loadings for HDL cholesterol and the rate of insulin-mediated glucose disposal (M, in milligrams per kilogram of body weight per minute). M also loaded on factor 2, along with fasting insulin and glucose concentrations, diastolic blood pressure, and waist girth. The observation that M loaded on both factors suggests that a resistance to insulin action may provide the mechanism uniting the features of the insulin resistance syndrome. Hyperinsulinemia with concomitant insulin resistance may be necessary to produce this metabolic derangement, as well as the increased risk of macrovascular complications. PMID- 9409210 TI - The heterozygous 20210 G/A prothrombin genotype is associated with early venous thrombosis in inherited thrombophilias and is not increased in frequency in artery disease. AB - A genetic variation in the 3'-untranslated region of the prothrombin mRNA (20210 G/A) has recently been reported to be associated with elevated plasma prothrombin levels and with an increased incidence of venous thrombosis. We determined the frequency of this mutation, the detection of which was improved by allele specific amplification of exon 14 and by denaturing gradients (denaturing gradient gel electrophoresis), in cohorts of patients affected by venous thrombosis (n = 132) or by coronary or cerebrovascular diseases (n = 195) and in normal subjects from various populations. An overlapping frequency of the heterozygous genotype (4%) was found in normal subjects from Italy and Cyprus, and no carrier was detected in 40 subjects of Indian or Somali origin. The 20210 GA heterozygous genotype was not increased in frequency in patients with arterial disease. In contrast, the GA genotype was associated (P = .007) with venous thrombosis both in simple heterozygotes (16%) with a family history of thrombosis as well as in double heterozygotes (14%) for other known thrombophilic defects. A synergic interaction between the prothrombin 20210 GA genotype and the factor V Leiden mutation, both potentially affecting the prothrombinase complex, was suggested by the early onset of thrombosis (median age 22 years) in doubly heterozygous patients. The association of the 20210 A allele with higher prothrombin levels was confirmed in the Italian population. However, the prothrombin assay does not allow an efficient preselection of patients for the DNA analysis. PMID- 9409211 TI - Local application of LDL promotes intimal thickening in the collared carotid artery of the rabbit. AB - Oxidized LDL (oxLDL) has been implicated in atherogenesis on the basis of in vitro studies and is present in atherosclerotic lesions. The aim of this study was to investigate the effects of LDL and oxLDL on intimal thickening in vivo. Intimal thickening was evoked by the placement of silicone collars around the carotid arteries of rabbits for 2 weeks. The collars were connected to osmotic minipumps containing LDL (7 micrograms h-1, n = 16 arteries), oxLDL (Cu2+ oxidized, 7 micrograms h-1, n = 16), or phosphate-buffered saline (5 microL h-1, n = 16). Segments proximal to the collars served as controls. Collar placement without lipoprotein application resulted in the appearance of alpha-SMC actin immunoreactive cells in the intima, thereby increasing the intimal thickness from 5 +/- 1 to 26 +/- 5 microns. The perivascular infusion of LDL or oxLDL within the collar significantly enhanced the development of the intima ninefold and sevenfold, respectively. The large intimas resulting from lipoprotein exposure were infiltrated by macrophages and T lymphocytes, and the intimal collagen area was increased from 5 +/- 2% in the discrete collar-induced intima to approximately 20% in the lipoprotein-evoked lesions. In conclusion, the local vascular application of LDL, oxidized in vitro or possibly in vivo, elicited an inflammatory-fibroproliferative response characteristic of arteriosclerotic lesions, thereby demonstrating an active role for this class of lipoproteins in the disease process. PMID- 9409212 TI - Tissue prothrombin. Universal distribution in smooth muscle. AB - Immunohistochemical analysis of surgically obtained porcine tissue samples reveals ubiquitous staining for prothrombin in organs rich in smooth muscle content and universal staining of smooth muscle in tissue vasculature. The native character of tissue prothrombin is verified first by chromogenic substrate hydrolysis and hirudin inhibition after incubation of tissue extracts with taipan snake venom and phospholipid. Western analysis of tissue extracts confirms the native zymogen molecular weight. In addition, prothrombin purified in good yield from porcine uterus is activated by Echis carinatus venom which, like taipan venom, is 4-carboxyglutamic acid-sensitive. After correction for blood (gross heme) and interstitial fluid (albumin), excess functional prothrombin is observed in extracts of tissues having abundant smooth muscle. In contrast with factor X, the yield of prothrombin purified from porcine uterus greatly exceeds that attributable to contamination by whole blood. Northern blot analysis from selected bovine tissues extracted for polyadenylated messenger RNA is equivocal for prothrombin mRNA with the exception of liver, which is positive. It is concluded that functionally intact prothrombin is widely distributed among tissues owing to smooth muscle content, although the mechanism of emplacement and physiologic significance of prothrombin in these tissues remains unclear. PMID- 9409214 TI - Immunohistochemical colocalization of the terminal complex of human complement and smooth muscle cell alpha-actin in early atherosclerotic lesions. AB - There is substantial evidence that activated components of the complement cascade are present in atherosclerotic lesions, and it was suggested some years ago that smooth muscle cells may be an important target of complement attack by the terminal components of the cascade, C5b-9, also called the membrane attack complex. Recent in vitro studies have shown that assembly of membrane attack complex on smooth muscle cells leads to the release of monocyte chemotactic protein-1, and, if this were to occur in vivo, then it could be responsible for the recruitment of monocytes into the lesion. In this study we have investigated the localization of C5b-9 in early atherosclerotic lesions of human coronary arteries, collected from autopsies, by immunohistochemical staining, C5b-9 was found to colocalize widely with smooth muscle cell alpha-actin, but not with intact macrophages, thus supporting the hypothesis that interaction of complement with smooth muscle cells may indeed be important in atherogenesis. PMID- 9409213 TI - Identification of osteoglycin as a component of the vascular matrix. Differential expression by vascular smooth muscle cells during neointima formation and in atherosclerotic plaques. AB - Using differential cDNA screening, we demonstrated that the bone-associated glycoprotein osteoglycin was highly expressed in differentiated adult rat vascular smooth muscle cells (VSMCs) but downregulated in VSMCs that had undergone proliferation in vitro. Further experiments in vitro revealed that osteoglycin gene expression was downregulated by a number of cytokines expressed in vivo (often in association with vascular injury) including basic fibroblast growth factor, transforming growth factor-beta, platelet-derived growth factor, and angiotensin II. In the normal adult rat carotid artery, osteoglycin was expressed in both the media and adventitia. However, osteoglycin mRNA expression was substantially increased in the adventitia and neointima 14 days after balloon injury, implying a role for this protein in vessel remodeling. Northern analysis of mRNA from neonatal rat aortas demonstrated upregulation of osteoglycin mRNA at week 2, after VSMC proliferation had ceased and when matrix modeling was maximal. In situ hybridization studies in human coronary arteries showed that osteoglycin mRNA was expressed by normal medial VSMCs but was downregulated in a subset of intimal VSMCs. Osteoglycin was not expressed in the VSMCs of adventitial vessels but was expressed in a subset of adventitial cells. This expression pattern contrasted with that of SM22 alpha, a contractile protein marker of VSMC differentiation, which was highly expressed in the media of all vessels. These data indicate that osteoglycin is a new marker of differentiated VSMCs and may be an essential component of the normal vascular matrix. PMID- 9409215 TI - Adenovirus-mediated expression of the secreted form of basic fibroblast growth factor (FGF-2) induces cellular proliferation and angiogenesis in vivo. AB - Blood supply through collateral arteries is of critical importance in occlusive arterial diseases such as coronary atherosclerosis. Induction of angiogenic growth factor within either the narrowing arteries or jeopardized myocardium may promote angiogenesis in vivo, leading to salvage of ischemic myocardium. We constructed a replication-defective adenovirus (AdCAsFGF-2) coding for human basic fibroblast growth factor (FGF)-2 that is modified, so that its secretion will be facilitated, by tagging a signal sequence derived from FGF-4. A large quantity of FGF-2 was detected in both the cell lysate and culture medium of COS cells infected with AdCAsFGF-2, indicating that FGF-2 was secreted at least partly from the infected cells. The conditioned medium from the infected COS cells stimulated DNA synthesis in and induced cellular proliferation of arterial smooth muscle cells. These effects were eliminated by adenovirus-mediated overexpression of a dominant-negative truncated FGF-receptor type 1. Implantation of a gel of basement membrane proteins containing fibroblasts infected with AdCAsFGF-2 into the ventral subcutaneous space of mice induced extensive cellular proliferation and the formation of functional arterioles. Cells surrounding the vessels were positively immunostained with antibodies recognizing either smooth muscle-specific alpha-actin or factor VIII antigen as a marker for endothelium. These results suggest that AdCAsFGF-2 may be useful for delivering functional FGF 2 into tissues and may lead to therapeutic angiogenesis in vivo. PMID- 9409216 TI - Inhibition of protein tyrosine kinases attenuates increases in expression of transforming growth factor-beta isoforms and their receptors following arterial injury. AB - Transforming growth factor-beta 1 (TGF-beta 1) has been implicated in neointima formation in mechanically injured vessels and in restenosis after angioplasty. To further understand the significance of TGF-beta s in neointima formation, we examined the temporal expression of three TGF-beta isoforms (-beta 1, -beta 2, and -beta 3), their receptors (ALK-2, ALK-5, and T beta RII), and two putative TGF-beta responses (elevations in alpha v and beta 3 integrin mRNAs) in balloon catheter-injured rat carotid arteries and their dependency on tyrosine kinase activity. Using a standardized reverse transcriptase-polymerase chain reaction assay optimized to estimate mRNA levels, we observed distinct patterns of mRNA regulation for TGF-beta 1, -beta 2, and -beta 3 during the 48 hours immediately after injury, which were localized to the vessel's media. TGF-beta 1 mRNA increased 10-fold during this time while TGF-beta 3 mRNA also increased almost 2 fold. There were also increases in mRNAs encoding the TGF-beta type I receptors ALK-5 and ALK-2, as well as the type II receptor (T beta RII). Eight hours after the injury, mRNA levels for ALK-2 and ALK-5 were on average 2-fold higher; mRNA encoding the type II receptor increased approximately 3-fold by 24 hours. There were also associated increases in TGF-beta 1, TGF-beta 3, ALK-5, and T beta RII immunoreactive peptide levels. Peak increases in mRNAs for integrins alpha v and beta 3 averaged approximately 2-fold and 2.5-fold, respectively. Perivascular administration of the tyrosine kinase inhibitor genistein at the time of vessel injury markedly (> 85%) inhibited elevations in mRNAs encoding TGF-beta 1, TGF beta 3, T beta RII, and the two integrins alpha v and beta 3, while application of its inactive chemically similar homologue daidzein did not prevent the injury induced elevations in mRNA levels. Since the increases in integrins alpha v and beta 3 mRNA could be theoretically attributed to TGF-beta actions despite being dependent on tyrosine kinase activity, we examined whether the observed elevations in integrins alpha v and beta 3 were due to TGF-beta 1 secretion, using cultured rat carotid artery smooth muscle cells. TGF-beta 1 neutralizing antibodies specifically inhibited elevations in integrins alpha v and beta 3 mRNAs due to platelet-derived growth factor-BB and fibroblast growth factor-2. We conclude that multiple components of the TGF-beta system in vessels are activated following injury and influence expression of integrin receptors important for smooth muscle cell migration. Activation of the TGF-beta system appears to be highly dependent on tyrosine kinases. PMID- 9409217 TI - Basic fibroblast growth factor (bFGF) regulates the expression of the CC chemokine monocyte chemoattractant protein-1 (MCP-1) in autocrine-activated endothelial cells. AB - The CC chemokine monocyte chemoattractant protein (MCP)-1 is induced by inflammatory cytokines and acts as a potent regulator of monocyte trafficking. Monocytes adhere preferentially to migrating endothelial cells in vitro and to endothelial cells at the migration front in vivo after aortic balloon denudation injury. Based on these findings, we analyzed MCP-1 expression in migrating and resting bovine aortic endothelial (BAE) cells and identified prominently upregulated levels of MCP-1 expression in migrating BAE cells. Stimulation of resting BAE cells with 5 ng/mL bFGF resulted in a fourfold induction of MCP-1 mRNA expression. The time course of bFGF-induced MCP-1 mRNA expression indicated a rapid and direct stimulation of MCP-1 expression that was detectable 30 minutes after stimulation. Levels of basal MCP-1 expression, as well as upregulated levels of MCP-1 in migrating BAE cells, were downregulated by addition of a neutralizing anti-bFGF monoclonal antibody (1.0 microgram/mL). Digestion of conditioned media of resting BAE cells with collagenase led to a dose-dependent induction of MCP-1 expression in resting BAE cells, which was inhibited > 50% by addition of neutralizing anti-bFGF antibody. Confirmation of the Northern blot experiments by ELISA-based quantitation of MCP-1 protein levels identified threefold to sixfold higher levels of MCP-1 in the supernatants of bFGF stimulated BAE cells than in unstimulated resting BAE cells. Finally, analysis of MCP-1 expression by in situ hybridization carried out on en face preparations of aortas demonstrated that MCP-1 expression is dramatically upregulated in regenerating endothelial cells in vivo after balloon denudation. Though not establishing a direct causal relation between the preferential adhesion of monocytes to migrating endothelial cells, these findings strongly suggest that autocrine-activated endothelial cell-derived MCP-1 may play a critical role in recruiting monocytes. They furthermore support the concept that bFGF acts as an autocrine regulator of endothelial cell activity and may imply an involvement of bFGF as a mediator of inflammatory cell trafficking. PMID- 9409218 TI - Expression of multiple isoforms of nitric oxide synthase in normal and atherosclerotic vessels. AB - Atherosclerosis is associated with reduced endothelium-derived relaxing factor bioactivity. To determine whether this is due to decreased synthesis of nitric oxide synthase (NOS), we examined normal and atherosclerotic human vessels by in situ hybridization and immunocytochemistry by using probes specific for endothelial (ecNOS), inducible (iNOS), and neuronal (nNOS) NOS isoforms, ecNOS was detected in endothelial cells overlying normal human aortas, fatty streaks, and advanced atherosclerotic lesions. A comparison of the relative expression of ecNOS to von Willebrand factor on serial sections of normal and atherosclerotic vessels indicated that there was a decrease in the number of endothelial cells expressing ecNOS in advanced lesions. iNOS and nNOS were not detected in normal vessels, but widespread production of these isoforms was found in early and advanced lesions associated with macrophages, endothelial cells, and mesenchymal appearing intimal cells. These data suggest that there is (1) a loss of ecNOS expression by endothelial cells over advanced atherosclerotic lesions and (2) a significant increase in overall NOS synthesis by other cell types in advanced lesions composed of the ecNOS, nNOS, and iNOS isoforms. We hypothesize that the increased expression of NOS and presumably NO in atherosclerotic plaques may be related to cell death and necrosis in these tissues. PMID- 9409219 TI - Smooth muscle cells express granulocyte-macrophage colony-stimulating factor in the undiseased and atherosclerotic human coronary artery. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF), one of a family of cytokines that regulate proliferation in macrophages and other types of cells, has been implicated in the inflammatory-fibroproliferative response of atherosclerosis. However, previous studies have been restricted to cultured cells and animal models. In the present study, we investigated GM-CSF expression in undiseased and atherosclerotic human coronary arteries at both the mRNA and protein levels. Dual in situ hybridization/cell-marking experiments demonstrated that subpopulations of intimal smooth muscle cells (SMCs) and endothelial cells express the cytokine in the histologically normal human coronary artery and that augmented expression occurs at these sites, and in macrophage accumulations and medial SMCs, in the atherosclerotic vessel. Corresponding data were obtained by in situ hybridization and reverse transcription-polymerase chain reaction and Northern analyses of cultured cells. Cultured human coronary arterial SMCs showed constitutive expression of GM-CSF in cells that had adopted an activated synthetic phenotype. Electron microscope immunocytochemistry revealed that GM-CSF is a protein localized in the cytoplasmic matrix of SMCs of both the undiseased and atherosclerotic vessel wall; extracellular matrix was largely unlabeled, with only occasional small patches of amorphous immunopositive material. The expression of GM-CSF by subpopulations of intimal SMCs in the undiseased artery and the marked upregulation of GM-CSF apparent in atherosclerotic lesions suggest roles for the cytokine in the cellular events underlying initiation and progression of the human atherosclerotic lesion. PMID- 9409220 TI - Short-term exposure to thapsigargin inhibits neointima formation in human saphenous vein. AB - Vascular smooth muscle cell (VSMC) migration and proliferation are involved in the intimal thickening responsible for late vein graft failure. In addition to growth and chemotactic factors, VSMCs require expression of matrix-degrading enzymes, e.g., metalloproteinases (MMP), to relieve the antiproliferative and antimigratory constraints of the extra-cellular matrix. Thapsigargin irreversibly inhibits Ca(2+)-ATPase, eliciting an increase in intracellular Ca2- and depletion of the intracellular calcium pools that are thought to be involved in the control of VSMC migration, VSMC proliferation, and MMP activity. We therefore studied the effect of thapsigargin on VSMC migration, VSMC proliferation, and MMP expression in human saphenous vein organ cultures. Vein segments were cultured for 14 days, and VSMC proliferation and migration were determined by autoradiography. Cell death was assessed using in situ end-labeling and lactate dehydrogenase release. Using Western blotting, we examined MMP-2 and MMP-9 and tissue inhibitor of metalloproteinases (TIMP)-1 and TIMP-2 expression. Exposure to thapsigargin at 10 nmol/L for 60 minutes before culture significantly inhibited neointimal thickening (60%, P < .05), intimal and medial VSMC proliferation (32%, P < .05 and 37%, P < .05, respectively), and VSMC migration (36%, P < .05). Thapsigargin at 10 nmol/L did not significantly increase cell death or MMP-2, MMP-9, TIMP-1, and TIMP-2 expression. These results suggest that blockade of Ca(2+)-ATPase by thapsigargin inhibits VSMC migration and proliferation involved in neointimal formation without affecting MMP-2 and MMP-9 expression. Because short-term exposure to thapsigargin was sufficient to inhibit neointima formation, this drug may prove useful in the treatment of intimal thickening after arterial bypass graft surgery. PMID- 9409222 TI - Dose-dependent suppression of transplant arteriosclerosis in aorta-allografted, cholesterol-clamped rabbits. Suppression not eliminated by the cholesterol raising effect of cyclosporine. AB - Cyclosporine may suppress transplant arteriosclerosis; however, it also raises plasma cholesterol, which could promote the disease. Our aim was to test these hypotheses experimentally. In experiment 1 (n = 34), cholesterol was clamped at a human level of 5 to 7 mmol/L, and rabbits were given either saline or cyclosporine in a low, medium or high dose. In experiment 2 (n = 15), in which dietary cholesterol was fixed at 0.05 g.kg-1.d-1, and experiment 3 (n = 16), in which no dietary cholesterol was added to the chow, rabbits were given either medium-dose cyclosporine, saline, or vehicle. The duration of each experiment was 5 weeks. In experiment 1, cyclosporine attenuated the development of transplant arteriosclerosis dose dependently (trend test: P < .0001). Cyclosporine also suppressed, in a dose-dependent manner, the activation of the immune system (trend test: P < .05) and the presence of T lymphocytes (trend test: P < .0001) and macrophages in the intima (trend test: P < .01). Despite a higher plasma cholesterol level in cyclosporine-treated rabbits compared with saline-treated rabbits in both experiment 2 (4.9 versus 2.9 mmol/L) and experiment 3 (1.6 versus 0.8 mmol/L), transplant arteriosclerosis was significantly reduced by cyclosporine (Mann-Whitney U test; P < .05 and P < .05). These results suggest that cyclosporine suppresses experimental transplant arteriosclerosis dose dependently. Accordingly, in the assessment of the optimal cyclosporine dose to heart-transplanted patients, it should be taken into account that a dose reduction may promote transplant arteriosclerosis. PMID- 9409221 TI - Interaction of very-low-density, intermediate-density, and low-density lipoproteins with human arterial wall proteoglycans. AB - The specific interaction of lipoproteins with arterial wall constituents, particularly proteoglycans (APG), is believed to play an important role in the development of atherosclerosis. The objective of this study was to examine the interaction of apolipoprotein B (apoB) containing lipoprotein subfractions (VLDL1, Sf 60 to 400; VLDL2, Sf 20 to 60; IDL1, Sf 16 to 20; IDL2, Sf 12 to 16; LDLA, Sf 8 to 12; and LDLB, Sf 0 to 8) prepared by cumulative density gradient centrifugation with chondroitin sulfate-rich APG. Eighteen subjects were studied, and a similar pattern of interaction between the lipoprotein species and APG was found in all. The order of reactivity (as measured by increased turbidity due to insoluble complex formation) was IDL Sf 12 to 16 > or = LDL Sf 8 to 12 > LDL Sf 0 to 8 > IDL Sf 16 to 20 >> VLDL Sf 20 to 60 > VLDL Sf 60 to 400. When the subjects were divided on the basis of their LDL subfraction profile, the extent of insoluble complex formation was highest in the group in which small, dense LDLIII was predominant; intermediate in the group whose LDL was mainly LDLII; and lowest in the group with a high proportion of LDLI (the mean reactivity, AU at 600 nm. of APG with IDL Sf 12 to 16 and LDL Sf 8 to 12 was 0.66; 0.62 and 0.46, 0.43 and 0.20, and 0.21 for the three groups, respectively). Fibrate lipid-lowering treatment decreased the percentage of LDLIII and increased the percentage of LDLI within total LDL and reduced the reactivity of all apoB-containing lipoprotein fractions toward APG. Sialic acid content varied in different lipoprotein subfractions, being the highest in VLDL and lowest in LDL. However, across lipoprotein species, it did not significantly correlate with APG-binding reactivity, suggesting that other factors are important in determining the interaction of lipoproteins with APG. Modification of LDL arginine and lysine residues abolished the ability of the lipoprotein to interact with APG, a finding that supports the hypothesis that the interaction is dependent on key positively charged amino acids on apoB. These findings demonstrate that (1) the overall reactivity of apoB-containing lipoproteins is greatest in individuals with small, dense LDL and (2) within an individual, IDL of Sf 12 to 16 is the most reactive species, and this may in part explain the positive correlation between IDL and risk of coronary heart disease. PMID- 9409223 TI - Soy protein versus soy phytoestrogens in the prevention of diet-induced coronary artery atherosclerosis of male cynomolgus monkeys. AB - Soy protein, long recognized as having cardiovascular benefits, is a rich source of phytoestrogens (isoflavones). To distinguish the relative contributions of the protein moiety versus the alcohol-extractable phytoestrogens for cardiovascular protection, we studied young male cynomolgus macaques fed a moderately atherogenic diet and randomly assigned to three groups. The groups differed only in the source of dietary protein, which was either casein/lactalbumin (casein, n = 27), soy protein with the phytoestrogens intact (soy+, n = 27), or soy protein with the phytoestrogens mostly extracted (soy-, n = 28). The diets were fed for 14 months. Animals fed soy+ had significantly lower total and LDL plus VLDL cholesterol concentrations compared with the other two groups. They soy+ animals had the highest HDL cholesterol concentrations, the casein group had the lowest, and the soy- group was intermediate. A subset was necropsied for atherosclerosis evaluations (n = 11 per group). Morphometric and angiochemical measures were done to quantify atherosclerosis. Coronary artery atherosclerotic lesions were smallest in the soy+ group (90% less coronary atherosclerosis than the casein group and 50% less than the soy- group), largest in the casein group, and intermediate in the soy- group. The effects of the diets on lesion size and arterial lipid measures of the peripheral arteries were similar to those in the coronary arteries, with greatest prevention of atherogenesis with soy+ and intermediate benefit with soy- relative to casein. We could not determine whether the beneficial effects seen in the soy- group relate to the protein itself or to the remaining traces of phytoestrogens. The beneficial effects of soy protein on atherosclerosis appear to be mediated primarily by the phytoestrogen component. Testicular weights were unaffected by the phytoestrogens. PMID- 9409224 TI - Correction of hypertriglyceridemia and impaired fat tolerance in lipoprotein lipase-deficient mice by adenovirus-mediated expression of human lipoprotein lipase. AB - Humans homozygous or heterozygous for mutations in the lipoprotein lipase (LPL) gene demonstrate significant disturbances in plasma lipoproteins, including raised triglyceride (TG) and reduced HDL cholesterol levels. In this study we explored the feasibility of adenovirus-mediated gene replacement therapy for LPL deficiency. A total of 5 x 10(9) plaque-forming units (pfu) of an E1/E3-deleted adenovirus expressing either human LPL (Ad-LPL) or the bacterial beta galactosidase gene (Ad-LacZ) as a control were administered to mice heterozygous for targeted disruption in the LPL gene (n = 57). Peak expression of total postheparin plasma LPL activity was observed at day 7 in Ad-LPL mice versus Ad LacZ controls (834 +/- 133 vs 313 +/- 89 mU/mL, P < .01), and correlated with human-specific LPL activity (522 +/- 219 mU/mL) and mass (9214 +/- 782 ng/mL), a change that was significant to 14 and 42 days, respectively. At day 7, plasma TGs were significantly reduced relative to Ad-LacZ mice (0.17 +/- 0.07 vs 1.90 +/- 0.89 mmol/L, P < .01) but returned to endogenous levels by day 42. Ectopic liver expression of human LPL was confirmed by in situ hybridization analysis and from raised LPL activity and mass in liver homogenates. Analysis of plasma lipoprotein composition revealed a marked decrease in VLDL-derived TGs. Severely impaired oral and intravenous fat-load tolerance in LPL-deficient mice was subsequently corrected after Ad-LPL administration and closely paralleled that observed in wild-type mice. These findings suggest that liver-targeted adenovirus-mediated LPL gene transfer offers an effective means for transient correction of altered lipoprotein metabolism and impaired fat tolerance due to LPL deficiency. PMID- 9409225 TI - Beneficial effects of alcohol withdrawal on LDL particle size distribution and oxidative susceptibility in subjects with alcohol-induced hypertriglyceridemia. AB - LDL subclass pattern B, reported to have a higher prevalence in hypertriglyceridemics (HTGs), is considered to be associated with an increased risk for coronary artery disease, and the small dense LDL characteristic of this pattern is susceptible to oxidative modification. Alcohol is considered one of the most frequent causes of increases in plasma triglyceride (TG) levels. We investigated the effects of alcohol withdrawal on LDL subclass distribution and oxidizability in drinkers with different plasma TG levels. Thirty-seven male subjects with relatively heavy alcohol-consumption habits were divided into four groups; normotriglyceridemic (NTG)/withdrawal (n = 11), NTG/control (n = 8), hypertriglyceridemic (HTG)/withdrawal (n = 10), and HTG/control (n = 8). Both withdrawal groups abstained form alcohol for 4 weeks, while the control subjects maintained their usual intake of alcohol. Peak LDL particle diameter (PPD) was smaller in the combined HTG groups than in the combined NTG groups before abstinence, although PPD increased significantly (P < .01) from 25.5 to 26.1 nm in the HTG/withdrawal group. Before abstinence, lag times preceding LDL oxidation in the combined HTG groups were shorter than in the combined NTG groups; after withdrawal, lag time was prolonged significantly (P < .01) from 49.9 to 57.3 minutes in the HTG-withdrawal group. No significant changes in PPD and lag time were observed in the other three groups. Significant correlations (P < .05) were observed between the change (delta) in the lag time and delta TG and between delta lag time and delta PPD. We conclude that in alcohol-induced HTG subjects, alcohol withdrawal has beneficial effects on the LDL profile by shifting the particle size from smaller to larger and decreasing its susceptibility to oxidation. PMID- 9409226 TI - Contribution of factor VII genotype to activated FVII levels. Differences in genotype frequencies between northern and southern European populations. AB - The relationship between coagulation factor VII (FVII) levels in plasma and FVII genotypes, determined by three polymorphisms (5'F7, IVS7, and 353R/Q), were studied in 500 control subjects enrolled in European multicenter study. The selection of particular FVII genotypes and the analysis of variance clearly indicated the independent contribution of a single 5'F7 insertion (A2) or 353Q (M2) allele to lowering plasma levels of activated FVII (FVIIa) (by a mean 25%). The M2 allele alone was found to make a major contribution to the genetically determined component of the FVIIa levels. Genotypes associated with low FVII levels were significantly rarer in the northern part of Europe (Oslo) than in the southern part (Rome, Murcia). The contribution made by the FVII genotype to the total variance of FVIIa levels was higher (30%) than that made to either FVII activity (25%) or FVII antigen (12%). Subjects with different FVII genotypes showed up to fivefold differences in mean FVIIa values, thus allowing attribution of a substantial part of the considerable interindividual variation to genetic variation, which may be of assistance in the interpretation of FVIIa levels on an individual basis. When FVII levels were adjusted by age and by triglyceride levels, the contribution of FVII genotypes to the FVII phenotypic variance was virtually unchanged. Taken together, these data indicate that in healthy control subjects the FVII genotype is a major predictor of plasma FVIIa levels and would support further study on the role of FVII genetic components in the development of cardiovascular disease. PMID- 9409227 TI - Elevated fasting total plasma homocysteine levels and cardiovascular disease outcomes in maintenance dialysis patients. A prospective study. AB - There is an excess prevalence of hyperhomocysteinemia in dialysis-dependent end stage renal disease (ESRD) patients. Cross-sectional studies of the relationship between elevated total homocysteine (tHcy) levels and prevalent cardiovascular disease (CVD) in this patient population suffer from severe methodologic limitations. No prospective investigations examining the association between tHcy levels and the subsequent development of arteriosclerotic CVD outcomes among maintenance dialysis patients have been reported. To assess whether elevated plasma tHcy is an independent risk factor for incident CVD in dialysis-dependent ESRD patients, we studied 73 maintenance peritoneal dialysis or hemodialysis patients who received a baseline examination between March and December 1994, with follow-up through April 1, 1996. We determined the incidence of nonfatal and fatal CVD events, which included all validated coronary heart disease, cerebrovascular disease, and abdominal aortic/lower-extremity arterial disease outcomes. After a median follow-up of 17.0 months, 16 individuals experienced at least one arteriosclerotic CVD event. Cox proportional-hazards regression analyses, unadjusted and individually adjusted for creatinine, albumin, and total cholesterol levels, total/HDL cholesterol ratio, dialysis adequacy/residual renal function, baseline CVD, and the established CVD risk factors (ie, age, sex, smoking, hypertension, diabetes/glucose intolerance, and dyslipidemia) revealed that tHcy levels in the upper quartile (> or = 27.0 mumol/L) versus the lower three quartiles (< 27.0 mumol/L) were associated with relative risk estimates (hazards ratios, with 95% confidence intervals for the occurrence of (pooled) nonfatal and fatal CVD ranging from 3.0 to 4.4; 95% confidence intervals (1.1 8.1) to (1.6-12.2). We conclude that the markedly elevated fasting tHcy levels found in dialysis-dependent ESRD patients may contribute independently to their excess incidence of fatal and nonfatal CVD outcomes. PMID- 9409228 TI - Role of lipoprotein-bound NEFAs in enhancing the specific activity of plasma CETP in the nephrotic syndrome. AB - Plasma cholesteryl ester transfer protein (CETP) activity, evaluated by the transfer of radiolabeled cholesteryl esters from a tracer dose of tritiated HDL to the plasma apolipoprotein B-containing lipoproteins, was significantly higher in patients with untreated idiopathic nephrotic syndrome (n = 15) than in normolipidemic control subjects (n = 22) (81.5 +/- 8.4 versus 43.1 +/- 3.1 micrograms CE.mL-1.h-1, respectively; P < .001). The increased CETP activity in nephrotic plasma was explained by a significant rise in both the CETP mass concentration (3.2 +/- 0.2 versus 2.1 +/- 0.1 mg/L; P < .001), and the specific CETP activity, calculated as the ratio of CETP activity to CETP mass (25.3 +/- 1.7 versus 20.4 +/- 1.6 micrograms CE.mg-1.h-1; P < .05). Elevated CETP activity in nephrotic patients was shown to be associated with a significant decrease in the mean size of LDL (24.4 +/- 0.5 versus 26.3 +/- 0.5 nm; P < .0001) as well as in the relative abundance of HDL2a (29.6 +/- 1.6% versus 34.8 +/- 1.1%; P < .05). The nephrotic syndrome was characterized by a significant increase in the relative proportion of lipoprotein-bound nonesterified fatty acids (NEFAs) (35.4 +/- 7.7% versus 7.6 +/- 3.0% of total; P < .01), leading to a significant increase in the electronegative charge of LDL (-4.3 +/- 0.1 versus -3.9 +/- 0.1 mV; P < .05) and HDL (-11.5 +/- 0.1 versus -11.1 +/- 0.2 mV; P < .05). Compared with native, non-supplemented plasma, removal of lipoprotein-bound NEFAs by addition of fatty acid-poor albumin to total plasma from nephrotic patients or control subjects significantly decreased CETP activity and specific CETP activity. Specific CETP activity no longer differed between nephrotic and control groups after albumin supplementation (19.7 +/- 1.5 versus 17.7 +/- 1.5 micrograms CE.mg-1.h-1; NS). It is concluded that, in addition to elevated CETP mass concentration, lipoprotein-bound NEFAs, by increasing the negative electrostatic charge of nephrotic lipoproteins, can facilitate the CETP-mediated neutral-lipid transfer reaction in total plasma from nephrotic patients. PMID- 9409229 TI - Regulation of tumor necrosis factor alpha- and interleukin-1-beta-induced induced adhesion molecule expression in human vascular smooth muscle cells by cAMP. AB - This study investigates the hypothesis that the elevation of intracellular cAMP may affect cytokine-induced expression of adhesion molecules on human vascular smooth muscle cells. In cultured human smooth muscle cells from coronary arteries and saphenous veins, tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1 beta) induced the expression of intercellular adhesion molecule (ICAM 1) and vascular cell adhesion molecule (VCAM-1), whereas interferon-gamma (INF gamma) selectively stimulated the expression of ICAM-1. Adenylyl cyclase was stimulated either by the stable prostacyclin mimetic cicaprost or by forskolin. Adhesion molecules were detected by a cell surface enzyme immunoassay and the respective mRNA by reverse transcriptase polymerase chain reaction (rt-PCR). Cicaprost as well as forskolin significantly inhibited TNF-alpha- and IL-1 beta induced cell surface expression of ICAM-1 and VCAM-1. Semiquantitative rt-PCR measurements showed a marked decrease of TNF-alpha- and IL-1 beta-induced mRNA levels of both adhesion molecules after preincubation with cicaprost. The stability of TNF-alpha-induced ICAM-1 and VCAM-1 expression at mRNA and protein level was not altered by cicaprost. The IFN-gamma-induced increase of cell surface expression of ICAM-1 and the respective mRNA levels, however, were not significantly altered by elevation of intracellular cAMP. Basal and stimulated cAMP levels, measured by radioimmunoassay, did not differ in TNF-alpha- and IFN gamma-treated cells. The present results demonstrate that the expression of adhesion molecules on human smooth muscle cells induced by cytokines is differentially modulated by activation of adenylyl cyclase. PMID- 9409230 TI - Peroxidation of LDL from combined-hyperlipidemic male smokers supplied with omega 3 fatty acids and antioxidants. AB - The effects of marine omega-3 polyunsaturated fatty acids (FAs) and antioxidants on the oxidative modification of LDL were studied in a randomized, double-blind, placebo-controlled trial. Male smokers (n = 41) with combined hyperlipidemia were allocated to one of four groups receiving supplementation with omega-3 FAs (5 g eicosapentaenoic acid and docosahexaenoic acid per day), antioxidants (75 mg vitamin E, 150 mg vitamin C, 15 mg beta-carotene, and 30 mg coenzyme Q10 per day), both omega-3 FAs and antioxidants, or control oils. LDL and human mononuclear cells were isolated from the patients at baseline and after 6 weeks of supplementation. LDL was subjected to cell-mediated oxidation by the patients' own mononuclear cells, as well as to Cu(2+)-catalyzed and 2,2'-azobis-(2 amidinopropane hydrochloride) (AAPH)-initiated oxidation. Extent of LDL modification was measured as lag time, the formation rate of conjugated dienes (CDs), the maximum amount of CDs formed, formation of lipid peroxides, and the relative electrophoretic mobility of LDL on agarose gels. Dietary supplementation with omega-3 FAs increased the concentration of total omega-3 FAs in LDL and reduced the concentration of vitamin E in serum. The omega-3 FA-enriched LDL particles were not more susceptible to Cu(2+)-catalyzed, AAPH-initiated, or autologous cell-mediated oxidation than control LDL. In fact, enrichment with omega-3 FAs significantly reduced the formation rate of CDs when LDL was subjected to AAPH-induced oxidation. Supplementation with moderate amounts of antioxidants significantly increased the concentration of vitamin E in serum and increased the resistance of LDL to undergo Cu(2+)-catalyzed oxidation, measured as increased lag time, reduced formation of lipid peroxides, and reduced relative electrophoretic mobility compared with control LDL. Supplementation with omega-3 FAs/antioxidants showed oxidizability of LDL similar to that of control LDL and omega-3 FA-enriched LDL. In conclusion, omega-3 FAs neither rendered the LDL particles more susceptible to undergo in vitro oxidation nor influenced mononuclear cells' ability to oxidize autologous LDL, whereas moderate amounts of antioxidants protected LDL against oxidative modification. PMID- 9409231 TI - Inhibition of HMG-CoA reductase by atorvastatin decreases both VLDL and LDL apolipoprotein B production in miniature pigs. AB - In the present studies, the 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor atorvastatin was used to test the hypothesis that inhibition of cholesterol biosynthesis in vivo with a consequent reduction in the availability of hepatic cholesterol for lipoprotein synthesis, would (1) reduce very low density lipoprotein (VLDL) apolipoprotein B (apoB) secretion into the plasma, (2) reduce the conversion of VLDL apoB to LDL apoB, and (3) reduce LDL apoB direct synthesis. ApoB kinetic studies were carried out in six control miniature pigs and in six animals after 21 days of administration of atorvastatin (3 mg/kg per day). Pigs were fed a fat- (34% of calories; polyunsaturated to monounsaturated to saturated ratio, 1:1:1) and cholesterol- (400 mg/d cholesterol; 0.1%; 0.2 mg/kcal) containing pig chow-based diet. Atorvastatin treatment significantly reduced plasma total cholesterol, LDL cholesterol, total triglyceride, and VLDL triglyceride concentrations by 16%, 31%, 19%, and 28%, respectively (P < .01). Autologous 131I-VLDL, 125I-LDL, and [3H]leucine were injected simultaneously into each pig, and apoB kinetic data were analyzed using multicompartmental analysis (SAAM II). The VLDL apoB pool size decreased by 29% (0.46 versus 0.65 mg/kg; P = .002), which was entirely due to a 34% reduction in the VLDL apoB production rate (PR) (1.43 versus 2.19 mg/kg per hour; P = .027). The fractional catabolic rate (FCR) was unchanged. The LDL apoB pool size decreased by 30% (4.74 versus 6.75 mg/kg; P = .0004), which was due to a 22% reduction in the LDL apoB PR (0.236 versus 0.301 mg/kg per hour; P = .004), since the FCR was unchanged. The reduction in LDL apoB PR was primarily due to a 34% decrease in conversion of VLDL apoB to LDL apoB; however, this reduction was not statistically significant (P = .114). Hepatic apoB mRNA abundance quantitated by RNase protection assay was decreased by 13% in the atorvastatin-treated animals (P = .003). Hepatic and intestinal LDL receptor mRNA abundances were not affected. We conclude that inhibition of hepatic HMG-CoA reductase by atorvastatin reduces both VLDL and LDL apoB concentrations, primarily by decreasing apoB secretion into the plasma and not by an increase in hepatic LDL receptor expression. This decrease in apoB secretion may, in part, be due to a reduction in apoB mRNA abundance. PMID- 9409232 TI - Antiatherogenic effects of a novel lipoprotein lipase-enhancing agent in cholesterol-fed New Zealand white rabbits. AB - Following our report that administration of 4-diethoxyphosphorylmethyl-N-(4-bromo 2-cyanophenyl) benzamide (NO-1886) to rats elevated postheparin lipoprotein lipase (LPL) activity through an increase in the enzyme mass, we now investigate antiatherogenic effects of NO-1886 in cholesterol-fed New Zealand White rabbits. For 20 weeks, four groups of male rabbits received regular rabbit chow (the normal control), 0.25% cholesterol-containing chow (the control), and cholesterol chow supplemented with 0.5% and 1.0% NO-1886, respectively. Postheparin LPL activity at week 10 was raised by 30% in 0.5% of the NO-1886 group and 40% in 1.0% of the NO-1886 group compared with those in the control. The area under the curve of plasma cholesterol level was not different in three cholesterol-fed groups whereas the area under the curve of HDL cholesterol was approximately twofold greater in the two NO-1886 groups than in the control, and the area under the curve of plasma triglyceride was reduced to the level of the normal control. LPL activity was correlated with HDL cholesterol (r = .764, n = 18) and triglyceride (r = -.627, n = 18). Relative atheromatous area, aortic cholesterol, and triglyceride contents were reduced to approximately 25%, 60%, and 55%, respectively, of the control values by NO-1886 ingestion. Multiple regression analysis of LPL, HDL cholesterol, and triglyceride indicated that HDL cholesterol was the most powerful protector against aortic cholesterol accumulation, and triglyceride was the one to protect against the atheromatous area. We concluded that NO-1886 prevented the development of atherosclerosis through increasing LPL activity with a consequent increase in HDL cholesterol and a decrease in triglyceride without a significant influence of plasma cholesterol level. PMID- 9409234 TI - Compensatory enlargement in coronary and femoral arteries is related to neither the extent of plaque-free vessel wall nor lesion eccentricity. A postmortem study. AB - Arteries may demonstrate compensatory enlargement in response to plaque accumulation. It has been proposed that enlargement is achieved by the expansion of the nondiseased (plaque-free) vessel wall. In this study, we assessed this hypothesis. Post mortem, 32 atherosclerotic coronary arteries (left anterior descending, n = 10; left circumflex, n = 11; and right coronary, n = 11) and 54 atherosclerotic femoral arteries were pressure fixed. Cross sections (coronary arteries, n = 537; femoral arteries, n = 1602) were obtained for analysis every 2.5 mm for the coronary arteries and every 5.0 mm for the femoral arteries. From these cross sections, we determined the degree of remodeling and an eccentricity index. Finally, we measured the extent of plaque-free vessel wall. The plaque free vessel wall was defined as (1) no plaque present or (2) plaque thickness < 0.5 mm. A very weak, negative correlation was observed between the degree of remodeling and the extent of the plaque-free vessel wall (coronary arteries: no plaque r2 = .13, P < .01; < 0.5 mm plaque r2 = .15, P < .05; femoral arteries: no plaque r2 = .02, P < .01; < 0.5 mm plaque r2 = 0.04, P < .01). The degree of remodeling did not correlate with the eccentricity index (coronary arteries r2 = .002, P > .05 and femoral arteries r2 = .001, P > .05). Thus, compensatory enlarged segments did not reveal a larger circumference of plaque-free vessel wall compared with segments that failed to enlarge. This study provides no support for the hypothesis that nondiseased vessel-wall expansion is responsible for compensatory enlargement in atherosclerotic arteries. PMID- 9409233 TI - Thrombin stimulates syndecan-1 promotor activity and expression of a form of syndecan-1 that binds antithrombin III in vascular smooth muscle cells. AB - Vascular smooth muscle (VSM) cells express transmembrane proteoglycans of the syndecan gene family. We reported previously that the expression of syndecans by VSM cells is regulated by mitogens such as serum, platelet-derived growth factor, and basic fibroblast growth factor and that syndecan expression is induced after balloon injury in vivo. We now show that thrombin is a potent inducer of syndecan 1 expression in VSM cells. Transient transfection experiments with a rat syndecan 1 promoter construct demonstrated that thrombin stimulates transcription of the syndecan-1 gene. Syndecan expression in response to thrombin was not inhibited by downregulation of protein kinase C. Thrombin-induced syndecan-1 expression was dependent on tyrosine kinase activity. Calcium was necessary for syndecan-1 expression, but increasing the intracellular calcium levels was not sufficient to induce syndecan-1 expression. Analysis of antithrombin III (AT III) binding activity revealed that thrombin caused an increase in the synthesis of syndecan-1 molecules that exhibited high-affinity AT III binding. These results suggest that VSM cells could play an important role in controlling local thrombus formation subsequent to vascular injury, via a feedback mechanism that involves thrombin induced stimulation of an inhibitor of thrombin activity. PMID- 9409235 TI - Platelet-derived growth factor beta-receptors can both promote and inhibit chemotaxis in human vascular smooth muscle cells. AB - The effect of the three platelet-derived growth factor (PDGF) isoforms AA, AB, and BB on migration was investigated in cultured human saphenous vein smooth muscle cells. The modified Boyden chamber technique yielded efficacies BB >> AB, AA = 0. However, the BB concentration-response relationship displayed a pronounced peak, occurring between 1 and 10 ng/mL, with no response above this range. Checkerboard analysis showed that the promotion of migration at low concentrations was chemotactic in nature but that the downturn was independent of gradient. Furthermore, at high concentrations BB was able to prevent chemotaxis induced by fetal calf serum and epidermal growth factor (EGF). Experiments using low concentrations of BB in combination with high concentrations of AA to saturate PDGF alpha-receptors in the presence and absence of a neutralizing antibody to alpha-receptors revealed that alpha-receptor activation induced partial inhibition of chemotaxis but this did not account for the inhibition of migration by high concentrations of BB. Despite possessing no significant chemotactic action itself, high concentrations of the AB isoform completely inhibited BB induced chemotaxis. Taken together these results suggest that the chemotactic signal induced by PDGF is dominated by PDGF beta-receptors and switches from positive at low concentrations to negative at higher concentrations. Stimulation of DNA synthesis by the three isoforms (as measured by [3H] thymidine incorporation) yielded saturable responses for the AB and BB isoforms, with similar efficacy and weak or no response for the AA isoform. Concentration-dependent patterns of tyrosine phosphorylation of certain proteins mirrored the form of the chemotactic response and suggest one possible underlying regulatory mechanism to account for the disparity between PDGF-induced chemotaxis and DNA synthesis. PMID- 9409236 TI - Prevalence of double pre-beta lipoproteinemia in hyperlipidemic patients is influenced by gender, menopausal status, and ApoE phenotype. AB - Double pre-beta lipoproteinemia (DPBL) is a plasma lipoprotein phenotype characterized by the presence of two agarose gel electrophoretic populations of very low density lipoproteins (VLDLs, d < 1.006 g/mL), i.e., normal pre-beta migrating VLDL and slow pre-beta VLDL. Slow pre-beta VLDL represents remnant lipoproteins derived from the hydrolysis of triglyceride (TG)-rich lipoproteins (TRLs), and thus DPBL is a characteristic of plasma remnant lipoprotein accumulation. To determine the prevalence of DPBL in our lipid clinic population, patients (n = 2501) were selected who (1) had an unambiguous VLDL electrophoretic phenotype and could be classified as having either DPBL (DPBL+), beta-migrating VLDL (beta-VLDL +), or an absence of both (DPBL/beta-VLDL-/-) and (2) had hypercholesterolemia (HC: plasma cholesterol > or = 6.2 mmol/L, n = 1017), hypertriglyceridemia (HTG: plasma TG > or = 2.3 mmol/L but < 15 mmol/L, n = 554) or combined hyperlipidemia (HC + HTG, n = 930). Patients with TG < 2.3 mmol/L and cholesterol < 5.2 mmol/L acted as control subjects (n = 343). Using a commercially available agarose gel electrophoresis system, we identified 220 hyperlipidemic patients (8.8%) with DPBL (versus < 1% of control). The prevalence of DPBL was higher in (1) male than in female patients (10.7% versus 6.7%), (2) postmenopausal than in premenopausal females (7.3% versus 4.1%), and (3) patients with HC + HTG than in those with HTG or HC alone (15.8% versus 8.3% versus 2.7%, respectively). Patients with an epsilon 2 allele had a higher prevalence of DPBL; i.e., 26.9% of apoE 3/2 and 26.2% of apoE 4/2 patients had DPBL compared with 6.5%, 6.8%, and 7.4% of apoE 3/3, 4/3, and 4/4 patients, respectively. DPBL patients consistently had increased levels of VLDL-C and (LDL + HDL)-TG and decreased levels of LDL-C, and their plasma lipid profiles were intermediate between those of beta-VLDL+ and DPBL/beta-VLDL -/- patients. These results demonstrate that male sex, postmenopausal status in women, and the presence of an apoE 3/2 or apoE 4/2 phenotype are associated with an increased incidence of DPBL in hyperlipidemic patients. PMID- 9409237 TI - Effect of iron overload and iron deficiency on atherosclerosis in the hypercholesterolemic rabbit. AB - It has been suggested that iron plays an important role in the pathogenesis of atherosclerosis, primarily by acting as a catalyst for the atherogenic modification of LDL. Although some epidemiological data suggest that high stored iron levels are an independent risk factor for coronary artery disease and that iron has been detected in both early and advanced atherosclerotic lesions, the evidence is often contradictory and inconclusive. We used the New Zealand White rabbit to investigate the effects of iron overload (FeO) and iron deficiency (FeD) on atherosclerosis. Groups of 7 rabbits were either iron loaded by injections of iron dextran (FeO group), iron depleted by phlebotomy (FeD group), or given injections of saline (control group) for a total of 9 weeks. All rabbits were fed a chow diet containing 1% (wt/wt) cholesterol for the last 6 weeks of the study. Iron and antioxidant status and cholesterol levels were assayed in plasma before cholesterol feeding (week 3) and at the time that the rabbits were killed (week 9). In addition, the susceptibility of LDL to oxidation was measured and pathological examination of the aortic arch and thoracic aorta performed at the end of the study. FeD significantly decreased the levels of blood hemoglobin, serum iron, and transferrin saturation compared with controls. Conversely, FeO significantly increased transferrin Fe saturation. FeO but not FeD decreased plasma cholesterol levels compared with control animals both before (P < .05) and after (P = .055) cholesterol feeding. Neither FeO nor FeD had a significant effect on the levels of antioxidants and lipid peroxidation products in plasma and aortic tissue or on the susceptibility of LDL to ex-vivo oxidation. FeO significantly decreased aortic arch lesion formation by 56% compared with controls (P < .05), whereas FeD had no significant effect. These results indicate that in this animal model, FeO decreases rather than increases atherosclerosis, likely because iron dextran exerts a hypocholesterolemic effect. Our data do not support the hypotheses that elevation of Fe stores increases or that a reduction of Fe stores by phlebotomy decreases the risk of coronary artery disease. PMID- 9409238 TI - Soluble vascular cell adhesion molecule-1 as a biohumoral correlate of atherosclerosis. AB - Vascular cell adhesion molecule-1 (VCAM-1) is a protein expressed on the surface of activated endothelial cells and expressed in early atherosclerosis. Because part of the protein is shed in the circulation and can be detected in peripheral plasma [soluble (s) VCAM-1], we hypothesized that sVCAM-1 may be a circulating marker of the presence and severity of atherosclerosis in humans. We selected 11 patients with essential hypertension plus peripheral vascular disease (PVD) and matched them for age, gender, body mass index, and smoking habits with 11 patients with uncomplicated essential hypertension (UH) and 11 healthy controls. We evaluated plasma concentrations of sVCAM-1 along with those of the soluble form of two other endothelial leukocyte adhesion molecules [sE-selectin and s intercellular adhesion molecule-1 (sICAM-1)] and other markers of endothelial dysfunction/ damage [s-thrombomodulin, plasminogen activator inhibitor type I, and von Willebrand factor (vWF)]. We also measured insulin, glucose, fibrinogen, total and HDL cholesterol, and the urinary albumin excretion (UAE), which may also be related to atherosclerosis. Results of these assays were related to the echographic assessment of the maximum intima-media thickness (IMTmax) at the carotid bifurcation, as an index of atherosclerosis in the carotids. PVD patients had a clearly elevated IMTmax [2.7 (1.1-3.1) mm, median (range)] compared with both UH patients [1.2 (0.8-2.4) mm] and controls [1 (0.6-2) mm]. sVCAM-1 was clearly higher in PVD patients [990 (273-1808) ng/mL, median (range)] versus 340 (236-975) ng/mL in UH and 386 (204-835) ng/mL in controls, and it separated clinical categories better than sICAM-1, vWF, glucose, insulin, UAE, triglycerides, or total, LDL or HDL cholesterol, sVCAM-1 was also the best biohumoral correlate of IMTmax (R = .59; P < .001) in univariate analysis. Because many of the biohumoral variables assessed were mutually intercorrelated, they were entered in a multivariate analysis to assess their contribution in explaining IMTmax variability. sVCAM-1 remained the only independent predictor of IMTmax and totally abolished the contribution of other variables to IMTmax variability. Thus, sVCAM-1 is a good biohumoral correlate of overt atherosclerosis, independent of underlying hypertension, and may be an in vivo marker of endothelial activation. Its potential value as a surrogate for global risk assessment and its behavior in intervention studies remain to be determined. PMID- 9409239 TI - High prevalence of hyperhomocysteinemia and asymptomatic vascular disease in siblings of young patients with vascular disease and hyperhomocysteinemia. AB - Hyperhomocysteinemia (HHC) is associated with an increased risk of atherosclerotic vascular disease and may be inherited. Fasting and postmethionine HHC are independent risk factors that overlap to a limited extent. To study the familial occurrence of HHC, we investigated the prevalence of HHC (both fasting and after methionine) among 450 siblings of 167 consecutive young patients with vascular disease and postmethionine HHC. Furthermore, all subjects with postmethionine HHC (n = 125) were invited for noninvasive vascular testing; 101 (80.8%) agreed. Of those with a normal postmethionine plasma level (n = 325), we randomly selected 73 subjects for further studies; 53 agreed (72.6%). Thus, a total of 154 siblings underwent ultrasonography of the carotid arteries, measurement of ankle-brachial pressure indices at rest and after a treadmill exercise test, and exercise electrocardiographic stress testing. We observed HHC after methionine, fasting, or both, in 27.8% (95% CI, 23.7 to 31.9), 11.1% (CI, 8.2 to 14.0) and 8.7% (CI, 6.1 to 11.3) of the siblings. Abnormal peripheral, coronary, or carotid artery tests were observed in 35.7% (CI, 28.1 to 43.3), 7.1% (CI, 3.0 to 11.2), and 7.1% (CI, 3.0 to 11.2). Univariate and multivariate analyses revealed weak evidence of a relationship with homocysteine levels. In conclusion, we found a high prevalence of HHC and asymptomatic vascular disease in siblings of young patients with vascular (mainly peripheral arterial) disease and HHC. Our data raise the possibility that homocysteine does not play a major role in the early, asymptomatic phases of vascular disease, at least among siblings of young patients with vascular disease. PMID- 9409240 TI - Triggering of beta 1-integrin chain induces platelet adhesion to cultured endothelium. AB - We report here that platelets adhere to cultured endothelial cells (EC) on exposure to the integrin beta 1 activating monoclonal antibody (mAb) BV7. The effect of BV7 is exerted mostly on platelets rather than EC. BV7 does not induce platelet aggregation or 5-hydroxytyptamine (5-HT) release and does not increase platelet adhesion to matrix proteins. Another activating beta 1 mAb, Lia1/2, triggers an effect similar to BV7. Blocking antibodies to alpha 2 and beta 1, but not to other integrin chains, are able to inhibit BV7-mediated adhesion. Moreover, the effect of BV7 requires active cellular metabolism and is not inhibited by platelet treatment with aspirin, by the PAF receptor antagonist BN50730, the phosphokinase C inhibitor staurosporin, or by the cAMP or cGMP enhancers prostaglandin E1 and sodium nitroprusside, respectively. Finally, BV7 mediated adhesion was enhanced by the endoperoxide analogue U46619. These data describe a novel mechanism of platelet adhesion to endothelial cells. This adhesion pathway appears to be mediated by alpha 2 beta 1-integrin on platelets and a still-undefined endothelial counter receptor. PMID- 9409241 TI - Ethnic variation and in vivo effects of the -93t-->g promoter variant in the lipoprotein lipase gene. AB - Recently, a (t-->g) transition at nucleotide -93 in the lipoprotein lipase (LPL) gene promoter has been observed in Caucasians. Here, we have compared the frequency of the -93g carriers in three distinct populations (Caucasians, South African Blacks, and Chinese). The carrier frequency in the Caucasian population was 1.7% (4/232), which was in contrast to the South African Black population, which had a frequency for this allele of 76.4% (123/161) of the individuals tested. This transition was not observed in the Chinese population under study. Near complete linkage disequilibrium between the -93g and the previously described D9N mutation was observed in the Caucasian population but not in South African Blacks. To further assess the ancestral origins of these DNA changes, DNA haplotyping using a CA repeat 5' to these substitutions was performed. The -93t allele was associated with only a few specific dinucleotide repeat sizes. In contrast, the -93g allele occurred on chromosomes with many different repeat lengths. The broad distribution of repeats on -93g carrying chromosomes, their high frequency in the South African Black population, and the conservation of the -93g allele among different species may suggest that the -93g allele is the ancestral allele on which a transition to t and the D9N mutations arose. The very high frequency of the -93g allele distinct from the N9 allele in a cohort of Black South Africans allowed us to specifically assess the phenotypic effects of the -93g allele on lipids. Individuals homozygous for the g allele at -93 showed mildly decreased triglycerides compared with individuals homozygous for the t allele (1.14 +/- 0.66 mmol/L versus 0.82 +/- 0.3; P = .04). Thus, the -93g allele in this cohort is associated with low plasma triglyceride levels. PMID- 9409242 TI - 17 beta-estradiol prevents fatty streak formation in apolipoprotein E-deficient mice. AB - The reality of the atheroprotective effect of estrogens is still a matter of debate, and its unknown mechanisms could involve favorable changes in blood lipids and lipoproteins and/or direct action at the level of the arterial wall. We used the recently developed animal model of atherosclerosis constituted by apolipoprotein E-deficient mice in an attempt to clarify these issues. Male and female animals, fed a low-fat chow diet, were treated with increasing doses of 17 beta-estradiol (E2) after castration and compared with testosterone treated and uncastrated (intact) animals. Total serum cholesterol, LDL-cholesterol, and HDL cholesterol concentrations decreased under E2 treatment in each sex and were weakly correlated with lesion area. However, a highly significant correlation between lesion area and serum E2 levels also suggested a direct action of E2 on cells of the vascular wall. A dose-response curve analysis revealed that these activities were sex-dependent, with females being nearly twice as sensitive to E2 as males. It also revealed that the atheroprotective activity was recruited at higher E2 concentrations than those needed by other E2 target tissues such as uterus or functions such as apoA-1 and LDL production and/or clearance rates. PMID- 9409243 TI - HDL phospholipid content and composition as a major factor determining cholesterol efflux capacity from Fu5AH cells to human serum. AB - The relationships of cell cholesterol efflux to HDL phospholipid (PL) content and composition in human serum were analyzed in two groups of subjects selected on the basis of their HDL cholesterol (HDL-C) levels: a norm-HDL group (1.10 mmol/L < HDL-C < 1.50 mmol/L) and a high-HDL group (HDL-C > 1.75 mmol/L). In the high HDL group, the relative fractional efflux was significantly higher than in the norm-HDL group, and in both groups, fractional efflux was correlated with a number of lipoprotein parameters, the best correlation and the only one that remained significant after multivariate analysis being with HDL phospholipid (HDL PL). Analysis of the HDL-PL subclasses revealed that HDL in the high-HDL sera was enriched with phosphatidylethanolamine (HDL-PE) and relatively deficient in sphingomyelin (HDL-SM) compared with norm-HDL sera. Moreover, the fractional efflux values in the high-HDL group were negatively correlated with the proportion of HDL-PE (r = -.64, P < .0001) and positively correlated with the proportion of HDL-SM (r = .43, P < .01). Thus, this study provides evidence that HDL-PL concentration can be used to predict the capacity of serum to accept cellular cholesterol. Among the differences described between norm-HDL and high HDL sera, the variability in PE to SM ratio might reflect changes in serum cholesterol acceptors that modulate the first step of reverse cholesterol transport. PMID- 9409244 TI - Seasonal variations of rheological and hemostatic parameters and acute-phase reactants in young, healthy subjects. AB - The incidence of cardiovascular diseases is increased in winter months. Recent studies have shown seasonal changes in plasma viscosity, fibrinogen, and factor VII activity with elevated levels during winter. An increase in these factors generates a "hypercoagulable state," which may lead to a rise in cardiovascular morbidity and mortality. It has been suggested that an increase in upper respiratory infections might be the underlying cause for the raised acute-phase reactants, in particular fibrinogen, during the winter season. We investigated seasonal variations of 26 parameters, determining blood rheology and hemostasis in 16 healthy volunteers (8 men and 8 women) aged 20 to 41 years. They were seen at monthly intervals over a period of 1 year. Seasonal variation with peak fitted values in the winter months was found for plasma viscosity (P < .001 for the seasonal difference), red blood cell deformability (P < .001), whole blood viscosity (P < .001), hemoglobin (P < .001), hematocrit (P < .001), mean corpuscular volume (P = .001), platelet count (P = .01), alpha 1-glycoprotein (P < .001), fibrinogen (measured by immunonephelometry; P < .001), plasminogen activator inhibitor-1 (P = .002), LDL cholesterol (P = .003), and triglyceride levels (P < .001). HDL cholesterol (P < .001) and cortisol (P = .001) showed inverse seasonal patterns, with a maximum during summertime. No statistically significant seasonal variations were seen for red blood cell aggregation, complement factor C4, total cholesterol, ceruloplasmin, haptoglobin, white blood cell count, and plasminogen. These data do not support the hypothesis that increased morbidity and mortality from cardiovascular diseases during winter may be mainly attributable to increased synthesis of acute-phase proteins due to infections. The cause for the seasonal variations in rheological and hemostatic parameters remains unclear and should be studied in more detail. PMID- 9409245 TI - Phosphoproteins regulated by the interaction of high-density lipoprotein with human skin fibroblasts. AB - Interaction of HDL with cells activates protein kinase C (PKC), a process that may be important in stimulating efflux of excess cellular cholesterol. Here we report that HDL treatment of cholesterol-loaded fibroblasts increases 32P labeling of three acidic phosphoproteins. These phosphoproteins, called pp80, pp27, and pp18 based on apparent M(r) in kD, were also phosphorylated by acute treatment of cells with phorbol myristate acetate, suggesting that they are regulated in response to PKC activation. The HDL-stimulated phosphorylation of pp80 and pp18 was significant after only 30 seconds and was sustained for at least 30 and 120 minutes, respectively, while increased phosphorylation of pp27 was transient, reaching a maximum at 10 minutes. Both pp27 and pp18 were phosphorylated on serine/threonine residues, whereas pp80 was phosphorylated on serine/threonine and tyrosine residues. Immunoprecipitation studies suggested that pp80 is the myristoylated alanine-rich C kinase substrate protein, but the identities of pp27 and pp18 are unknown. HDL and trypsin-digested HDL stimulated phosphorylation of pp80 and pp27, while purified apoA-I, apoA-II, or apoE had no stimulatory effects, indicating that the active component in HDL was trypsin resistant and unlikely to be an apolipoprotein. Conversely, HDL, apoA-I, apoA-II, and apoE all stimulated pp18 phosphorylation, while trypsin-digested HDL had less effect, consistent with pp18's being responsive to HDL apolipoproteins. Treatment of cholesterol-depleted cells with apoA-I also stimulated phosphorylation of pp18, but only transiently. These results suggest that HDL interaction with cells activates diverse PKC-mediated pathways that target different phosphoproteins. Of these three phosphoproteins, only pp18 has a phosphorylation response consistent with its being involved in apolipoprotein-mediated lipid transport. PMID- 9409246 TI - Upregulation of low density lipoprotein receptor by gemfibrozil, a hypolipidemic agent, in human hepatoma cells through stabilization of mRNA transcripts. AB - Gemfibrozil reduces the plasmal levels of cholesterol and triglyceride in patients with hyperlipidemia by a mechanism that is not well understood. The present study evaluated the effect of gemfibrozil on the LDL receptor in human hepatoma cells compared with that of pravastatin, an inhibitor of 3-hydroxy-3 methylglutaryl coenzyme A reductase. Exposure to gemfibrozil, 40 mumol/L, for 3 days increased the binding of 125I-LDL to the surface of three lines of human hepatoma cell, HepG2, HuH7, and HLE by 1.5- to 2.0-fold. Similar findings were observed with pravastatin. Scatchard analysis with 125I-LDL indicated an increased number of LDL receptors on the cell surface of HepG2 cells when treated with gemfibrozil and pravastatin. However, the gemfibrozil-treated cells exhibited no increase in the binding of 125I-epidermal growth factor (EGF). Gemfibrozil increased the levels of LDL receptor mRNA and protein in HepG2 cells. The increase in LDL receptor activity induced by pravastatin was abolished by concomitant administration of mevalonic acid, 770 mumol/L. This effect was not seen with gemfibrozil, suggesting the mechanism differs for the two lipid lowering drugs. To determine whether this increase in mRNA was due to transcriptional activation, we prepared HepG2 cells transfected with an LDL receptor promoter-reporter construct that contained a sterol regulatory element. The expression of LDL receptor regulated by the sterol regulatory element was increased by pravastatin, but not by gemfibrozil. We evaluated the stability of the mRNA in the presence of actinomycin D to explain the increase in the LDL receptor mRNA. Gemfibrozil prolonged the half-life of the mRNA for LDL receptor but not that for the EGF receptor. Stabilization of the LDL receptor mRNA is suggested to be the novel mode of action of gemfibrozil. PMID- 9409247 TI - Relation of LDL size to the insulin resistance syndrome and coronary heart disease in American Indians. The Strong Heart Study. AB - Small, dense LDL has been shown to be associated with the insulin resistance syndrome and coronary heart disease (CHD). We examined the distribution of LDL size and phenotype within a population-based sample of American Indians to determine the relationships with prevalent CHD and to examine associations with hyperinsulinemia and other components of the insulin resistance syndrome. Data were available for 4505 men and women between 45 and 74 years of age who are members of 13 American Indian communities in three geographic areas. Diabetes, CHD, and CHD risk factors were assessed by standardized techniques, and LDL size was measured by gradient gel electrophoresis. LDL size was smaller in men than in women and in individuals with diabetes than in those without diabetes. In multivariate analysis, LDL size was significantly related to several components of the insulin resistance syndrome, including triglycerides (inversely) and HDL cholesterol (positively). Although univariate relations were positive, LDL size was not significantly related to fasting insulin concentrations or body mass index in the multivariate model. LDL size also showed no relationship to apolipoprotein E phenotype. When LDL size was compared in individuals with and without CHD, no significant differences were observed, either in nondiabetic or diabetic individuals. We conclude that LDL size is most strongly related to lipoprotein components of the insulin resistance syndrome, especially plasma triglycerides. However, in this population with low LDL, it is not related to cardiovascular disease. PMID- 9409248 TI - In vitro interactions of oxidatively modified LDL with type I, II, III, IV, and V collagen, laminin, fibronectin, and poly-D-lysine. AB - The accumulation of LDL in the arterial intima is considered a key event in atherogenesis. We investigated the binding of oxidized LDL (ox-LDL) to microtiter plates coated with type I or II collagen, laminin, fibronectin, or poly-D-lysine. Oxidation of LDL, 125I-LDL, or Eu(3+)-LDL was performed with CuCl2, varying the time of oxidation. Bound lipoprotein was assessed by counting radioactivity or fluorescence in the wells. Binding of highly ox-LDL in PBS followed the order: type I collagen > poly-D-lysine > type II collagen > laminin > fibronectin. Comparing various collagen types, the binding of ox-LDL followed the order: type I > type V and, type III > type IV > type II collagen. Binding of ox-LDL in PBS was dependent on an increase in negative charge of ox-LDL. Testing certain amino acids as competitors for binding of highly ox-LDL to type I collagen put lysine first, followed by arginine and histidine. On laminin, histidine competed most, followed by lysine and arginine. When studying the influence of Na+, K+, Ca2+, Mg2+ (equivalent to their concentrations in the interstitial fluid), native LDL, moderately ox-LDL, and highly ox-LDL showed the same affinity to type I collagen. However, a fivefold dilution of the buffer increased the affinity of moderately and highly ox-LDL 3.9- and 10-fold compared with native LDL. Application of the F(ab')2 from a monoclonal antibody to ox-LDL revealed a strong competition of the binding of highly ox-LDL to type II collagen (60%), laminin (35%), type I collagen (20%), and poly-D-lysine (15%), whereas the binding to fibronectin was not affected. PMID- 9409249 TI - Etiologic heterogeneity of hyperapobetalipoproteinemia (hyperapoB). Results from segregation analysis in families with premature coronary artery disease. AB - Hyperapobetalipoproteinemia (hyperapoB) is a common familial lipoprotein disorder associated with premature coronary artery disease (CAD). HyperapoB is characterized by an increased number of small, dense LDL particles. Patients with hyperapoB may be normotriglyceridemic (normoTG) or hypertriglyceridemic (hyperTG). We tested the hypothesis that a major locus controls the hyperapoB phenotype by using data from 1035 participants in 145 families enriched for premature CAD. Segregation analysis was conducted, and results suggest etiologic heterogeneity in these families. Families (n = 55) with one or more hyperTG hyperapoB individuals strongly supported mendelian recessive inheritance of hyperapoB. Under this mendelian model, individuals with the high-risk genotype had a baseline risk of 0.78, but parental and spouse's hyperapoB phenotypes did influence the probability of displaying hyperapoB. Low-risk genotypes had virtually no risk of displaying hyperapoB. The other subgroup of families (n = 72), in which all hyperapoB individuals were normoTG, did not show any clear pattern of inheritance. Eighteen families did not have any hyperapoB individual. In the 55 families with hyperTG hyperapoB, diabetes was more prevalent in hyperapoB individuals (18.3% of hyperTG hyperapoB individuals, 9.6% of normoTG hyperapoB individuals) than in normal individuals (4.9%). Both hyperTG hyperapoB and normoTG hyperapoB phenotypes were significant predictors for blood pressure in the 55 families, but not in the total population. These associations further suggest a link between hyperapoB and the small, dense LDL syndromes. This study successfully demonstrated mendelian inheritance of the hyperapoB phenotype and also suggested etiologic heterogeneity of hyperapoB. PMID- 9409251 TI - Reduced progression of atherosclerosis in apolipoprotein E-deficient mice following consumption of red wine, or its polyphenols quercetin or catechin, is associated with reduced susceptibility of LDL to oxidation and aggregation. AB - The effect of consuming red wine, or its major polyphenol constituents catechin or quercetin, on the development of atherosclerotic lesions, in relation to the susceptibility of plasma LDL to oxidation and to aggregation, was studied in atherosclerotic apolipoprotein E deficient (E degree) mice. Forty E degree mice at the age of 4 weeks were divided into four groups, 10 mice in each group, and were supplemented for up to 6 weeks in their drinking water with placebo (1.1% alcohol); catechin or quercetin (50 micrograms/d per mouse), or red wine (0.5 mL/d per mouse). Consumption of catechin, quercetin, or red wine had no effect on plasma LDL or HDL cholesterol levels. The atherosclerotic lesion area was smaller in the treated mice by 39%, 46%, and 48%, respectively, in comparison with E degree mice that were treated with placebo. In accordance with these findings, cellular uptake of LDL derived after catechin, quercetin, or red wine consumption was found to be reduced by 31%, 40%, and 52%, respectively. These results were associated with reduced susceptibility to oxidation (induced by different modes such as copper ions, free radical generator, or macrophages) of LDL isolated after red wine or quercetin and, to a lesser extent after catechin consumption, in comparison with LDL isolated from the placebo group. Similar results were obtained when LDL was preincubated in vitro with red wine or with the polyphenols prior to its oxidation. Even in the basal oxidative state (not induced oxidation), LDL isolated from E degree mice that consumed catechin, quercetin, or red wine for 2 weeks was found to be less oxidized in comparison with LDL isolated from E degree mice that received placebo, as evidenced by 39%, 48%, and 49% reduced content of LDL-associated lipid peroxides, respectively. This effect could be related to enhanced serum paraoxonase activity in the polyphenol-treated mice. LDL oxidation was previously shown to lead to its aggregation. The present study demonstrated that the susceptibility of LDL to aggregation was reduced in comparison with placebo-treated mice, by 63%, 48%, or 50% by catechin, quercetin, and red wine consumption, respectively, and this effect could be shown also in vitro. The inhibition of LDL oxidation by polyphenols could be related, at least in part, to a direct effect of the polyphenols on the LDL, since both quercetin and catechin were found to bind to the LDL particle via the formation of an ether bond. We thus conclude that dietary consumption by E degree mice of red wine or its polyphenolic flavonoids quercetin and, to a lesser extent, catechin leads to attenuation in the development of the atherosclerotic lesion, and this effect is associated with reduced susceptibility of their LDL to oxidation and aggregation. PMID- 9409250 TI - Enhanced endothelin-B-receptor-mediated vasoconstriction of small porcine coronary arteries in diet-induced hypercholesterolemia. AB - The coronary vasoconstrictor effects of endothelins, mediated by both endothelin ETA and ETB receptors, may be differentially altered in pathophysiological states associated with endothelial dysfunction and elevated endothelin levels. Experimental hypercholesterolemia is associated with coronary endothelial dysfunction and increased circulating endothelin concentrations. These studies were designed to test the hypothesis that experimental hypercholesterolemia is characterized by a differentially altered coronary contractile response to ETA- and ETB-receptor stimulation, in vitro. Pigs were fed either a normal or a high cholesterol diet for 10 to 13 weeks. Changes in the intraluminal diameter of pressurized small coronary arteries (< 481 +/- 25 microns in diameter) to cumulative concentrations (10(-10) to 10(-6) mol/L) of endothelin-1 (ET-1), and sarafotoxin 6c (S6c), a specific ETB-receptor agonist, were measured using a video dimension analyzer. The maximal contraction attained with ET-1 was greater than with S6c in both normal (86 +/- 7% versus 47 +/- 7%, P = .001) and hypercholesterolemic (77 +/- 6% versus 37 +/- 7%, P < .001) pigs. At 10(-10) mol/L, vessels from hypercholesterolemic pigs manifested greater contraction to both ET-1 (23 +/- 6% versus 8 +/- 3%, P = .02) and S6c (17 +/- 5% versus 4 +/- 2%, P = .02). Incubation of arteries from hypercholesterolemic pigs with BQ-788 (ETB-receptor antagonist), but not FR-139317 (ETA-receptor antagonist), altered the contractile response to ET-1 at 10(-10) mol/L. Removal of the endothelium abolished the difference in response to S6c between normal and hypercholesterolemic pigs. These studies demonstrate that experimental hypercholesterolemia is characterized by enhanced coronary vasoconstriction to endothelins in vitro, the mechanism of which is mediated mainly through the ETB receptor. Thus, the ETB receptor has a role in regulation of coronary artery tone in both the steady-state and pathophysiological states. PMID- 9409252 TI - Common genomic variation in the APOC3 promoter associated with variation in plasma lipoproteins. AB - We hypothesized that common genomic variation that affected the expression and/or function of the products of the APOC3, APOE, FABP2, and PON1 genes would be associated with variation in biochemical phenotypes in a previously unstudied human sample. We determined genotypes of functional genomic variants of APOC3, APOE, FABP2, and PON1 in 509 adult aboriginal Canadians from an isolated community in Northern Ontario. We tested for genotype associations with plasma lipoprotein traits. We found that (1) common variation at nucleotide -455 of the APOC3 promoter was associated with variation in plasma triglycerides (P = .006) and (2) common variation of APOE determining plasma isoforms of apo E was associated with variation in plasma apo B (P = .009). Analysis of subjects classed by APOC3 markers showed that homozygosity for presence of a C at nucleotide -455 and a T at nucleotide -482 was associated with significantly increased plasma triglycerides in both men and women. Furthermore, this allele was approximately twice as frequent in subjects within the highest quartile of plasma triglycerides as in subjects within the lowest quartile. Since the DNA variation detected by the APOC3 markers affects in vitro expression of the gene product, it is possible that the marker itself caused the associations. However, the associations could also have resulted from linkage disequilibrium with other functional variants in APOC3 or the closely linked APOA1 and/or APOA4 genes. PMID- 9409253 TI - The role of a triplet repeat sequence of the very low density lipoprotein receptor gene in plasma lipid and lipoprotein level variability in humans. AB - The biological role of the very low density lipoprotein receptor (VLDL-R) in humans is not yet elucidated. This cellular receptor binds apolipoprotein E (apoE)-containing lipoparticles and is mainly expressed in peripheral tissues. The VLDL-R gene contains a polymorphic triplet (CGG) repeat located 19 bp upstream of the initiation codon. We explored the allelic distribution of this repeat in 1384 subjects of European Caucasian origin, 609 of them surviving a myocardial infarction. Six alleles corresponding to 5, 6, 7, 8, 9, and 11 repeats were detected in this population. The alleles 5, 8, and 9 were the most frequent, with frequencies of 0.413, 0.275, and 0.292, respectively. No association was found between the VLDL-R polymorphism and myocardial infarction. In controls without lipid lowering treatment, a statistically significant interaction between VLDL-R genotype and apoE phenotype was found for plasma triglycerides (P < .04), suggesting a gene-gene interaction. There was also a main effect of the VLDL-R polymorphism on LpE:B and LpA-I. The VLDL-R 9 allele was associated with lower levels of plasma LpE:B (P < .05) and higher concentrations of plasma LpA-I (P < .01) than the other alleles. These results suggest that VLDL-R has a modest influence on circulating lipoproteins in humans. PMID- 9409254 TI - Protein C activation and factor Va inactivation on human umbilical vein endothelial cells. AB - The inactivation of factor Va was examined on primary cultures of human umbilical vein endothelial cells (HUVECs), either after addition of activated protein C (APC) or after addition of alpha-thrombin and protein C (PC) zymogen. Factor Va proteolysis was visualized by Western blot analysis using a monoclonal antibody (alpha HVaHC No. 17) to the factor Va heavy chain (HC), and cofactor activity was followed both in a clotting assay using factor V-deficient plasma and by quantitation of prothrombinase function. APC generation was monitored using the substrate 6-(D-VPR)amino-1-naphthalenebutylsulfonamide (D-VPR-ANSNHC4H9), which permits quantitation of APC at 10 pmol/L. Addition of APC (5 nmol/L) to an adherent HUVEC monolayer (3.5 x 10(5) cells per well) resulted in a 75% inactivation of factor Va (20 nmol/L) within 10 minutes, with complete loss of cofactor activity within 2 hours. Measurements of the rate of cleavage at Arg506 and Arg306 in the presence and absence of the HUVEC monolayer indicated that the APC-dependent cleavage of the factor Va HC at Arg506 was accelerated in the presence of HUVECs, while cleavage at Arg306 was dependent on the presence of the HUVEC surface. Factor Va inactivation proceeded with initial cleavage of the factor Va HC at Arg506, generating an M(r) 75,000 species. Further proteolysis at Arg306 generated an M(r) 30,000 product. When protein C (0.5 mumol/L), alpha thrombin (1 nmol/L), and factor Va (20 nmol/L) were added to HUVECs an APC generation rate of 1.56 +/- 0.11 x 10(-14) mol/min per cell was observed. With APC generated in situ, cleavage at Arg506 on the HUVEC surface is followed by cleavage at Arg306, generating M(r) 75,000 and M(r) 30,000 fragments, respectively. In addition, the appearance of two novel products derived from the factor Va HC are observed when thrombin is present on the HUVEC surface: the HC is processed through limited thrombin proteolysis to generate an M(r) 97,000 fragment, which is further processed by APC to generate an M(r) 43,000 fragment. NH2-terminal sequence analysis of the M(r) 97,000 fragment revealed that the thrombin cleavage occurs in the COOH-terminus of the intact factor Va HC since both the intact HC as well as the M(r) 97,000 fragment have the same sequence. Our data demonstrate that the inactivation of factor Va on the HUVEC surface, initiated either by APC addition or PC activation, follows a mechanism whereby cleavage is observed first at Arg506 followed by a second cleavage at Arg306. The latter cleavage is dependent on the availability of the HUVEC surface. This mechanism of inactivation of factor Va is similar to that observed on synthetic phospholipid vesicles. PMID- 9409255 TI - Moderate genetic influences on plasma levels of plasminogen activator inhibitor-1 and evidence of genetic and environmental influences shared by plasminogen activator inhibitor-1, triglycerides, and body mass index. AB - Both genes and environmental factors have been reported to influence plasma levels of plasminogen activator inhibitor-1 (PAI-1). However, the relative importance of genetic influences (i.e., heritability) on plasma PAI-1 levels has not yet been investigated. Furthermore, PAI-1 levels are correlated with body mass index (BMI) and triglycerides. These correlations could reflect genetic and/or environmental factors in common to PAI-1, triglycerides, and BMI. We applied multivariate genetic analysis methods to assess the relative importance of genetic and environmental influences on plasma PAI-1 levels and to test the significance of genetic and/or environmental influences shared by PAI-1, triglycerides, and BMI in 217 pairs of middle-aged and elderly twins, of whom 113 pairs were reared apart and 121 pairs were women. The heritability estimate for PAI-1 levels was 42%. Individual-specific environmental factors explained 36% of the variance for PAI-1 levels. The remaining variance of PAI-1 was explained by rearing and residual-familial environmental factors. Furthermore, a genetic correlation of 1.00 between PAI-1 and triglycerides, a rearing environmental correlation of 1.00 between PAI-1 and BMI, a residual-familial environmental correlation of 1.00 between PAI-1 and triglycerides, and a genetic correlation of 0.63 between PAI-1 and BMI, were found. In conclusion, the present results suggest that genetic influences on plasma PAI-1 are moderate. Genetic and shared rearing or residual-familial environmental factors shared by PAI-1, BMI, and triglycerides explain the phenotypic association between these measures. It appears that all the genetic influences for PAI-1 are more or less shared with those for triglycerides and BMI. PMID- 9409256 TI - Lipoprotein(a) interactions with lipid and nonlipid risk factors in early familial coronary artery disease. AB - An interaction between high plasma lipoprotein(a) [Lp(a)], unfavorable plasma lipids, and other risk factors may lead to very high risk for premature CAD. Plasma Lp(a), lipids, and other coronary risk factors were examined in 170 cases with early familial CAD and 165 control subjects to test this hypothesis. In univariate analysis, relative odds for CAD were 2.95 (P < .001) for plasma Lp(a) above 40 mg/dL. Nearly all the risk associated with elevated Lp(a) was found to be restricted to persons with historically elevated plasma total cholesterol (6.72 mmol/L [260 mg/dL] or higher) or with a total/HDL cholesterol ratio > 5.8. Nonlipid risk factors were also found to at least multiply the risk associated with Lp(a). When Lp(a) was over 40 mg/dL and plasma total/HDL cholesterol > 5.8, relative odds for CAD were 25 (P = .0001) in multiple logistic regression. If two or more nonlipid risk factors were also present (including hypertension, diabetes, cigarette smoking, high total homocysteine, or low serum bilirubin), relative odds were 122 (P < 1 x 10(-12)). The ability of nonlipid risk factors to increase risk associated with Lp(a) was dependent on at least a mildly elevated total/HDL cholesterol ratio. In conclusion, high Lp(a) was found to greatly increase risk only if the total/HDL cholesterol ratio was at least mildly elevated, an effect exaggerated by other risk factors. Aggressive lipid lowering in those with elevated Lp(a) therefore appears indicated. PMID- 9409258 TI - Long-term probucol treatment reverses the severity of myocardial injury in watanabe heritable hyperlipidemic rabbits. AB - We previously reported that administration of NO donors ameliorates the severity of myocardial injury in cholesterol-fed rabbits. We now evaluated the effects of probucol, a lipid-lowering antioxidant that can preserve endothelium-dependent relaxation (EDR), in the aortas of cholesterol-fed rabbits. We examined the effects of short-term (7 days) or long-term (24 weeks) administration of 1% probucol on the size of infarcts resulting from 30 minutes of coronary occlusion followed by reperfusion (for 48 hours) in Watanabe heritable hyperlipidemic (WHHL) rabbits. Infarcts in untreated WHHL rabbits were significantly larger than those in the rabbits receiving the long-term but not the short-term treatment with probucol (72.2 +/- 5.4%, 37.6 +/- 6.4%, and 66.7 +/- 3.5%, respectively). Long-term probucol treatment also significantly reduced myeloperoxidase activity in both ischemic and nonischemic myocardium and suppressed P-selectin expression in the coronary vasculature. No significant differences were observed in hemodynamic parameters during myocardial ischemia/reperfusion. Long-term probucol treatment significantly reduced the surface area of atherosclerotic plaque lesions in the aorta (24.4 +/- 3.8% vs 46.3 +/- 6.3, P < .05). Moreover, long term probucol treatment restored acetylcholine-induced EDR in aortic rings but did not affect sodium nitroprusside-induced relaxation. Finally, long-term probucol treatment resulted in significantly elevated cGMP levels in the aorta. These results indicate that long-term probucol treatment significantly ameliorates myocardial injury in heritable atherosclerotic rabbits, perhaps by reducing the accumulation of leukocytes in the myocardium and atherosclerotic vascular lesions. Thus, long-term administration appears to suppress the progression of atherosclerotic vascular disease in this animal model. PMID- 9409257 TI - Vascular endothelial growth factor/vascular permeability factor produces nitric oxide-dependent hypotension. Evidence for a maintenance role in quiescent adult endothelium. AB - In vitro studies suggest that vascular endothelial growth factor/vascular permeability factor (VEGF/VPF) may stimulate release of nitric oxide (NO) from endothelial cells. To investigate the hemodynamic consequences of recombinant VEGF/VPF administered in vivo, recombinant human VEGF/VPF was administered as a bolus dose of 500 micrograms to anesthetized (n = 6) or conscious (n = 5) New Zealand White rabbits, as well as anesthetized rabbits with diet-induced hypercholesterolemia (HC; n = 7). Anesthetized Yorkshire farm pigs (no specific dietary pretreatment) were studied before and after receiving 500 micrograms intravenous (IV; n = 5) or intracoronary (IC; n = 5) VEGF/VPF. In anesthetized, normal rabbits, mean arterial pressure (MAP) fell by 20.5 +/- 1.4% (P < .05 versus baseline) within 3 minutes after IV VEGF/VPF. Pretreatment with N omega nitro-L-arginine caused a significant inhibition of VEGF/VPF-induced hypotension. In conscious, normal rabbits, VEGF/VPF produced a consistent though lesser reduction in MAP. The fall in MAP induced by VEGF/VPF in anesthetized, HC rabbits (21.5 +/- 2.5% from baseline) was no different from that observed in normal anesthetized rabbits. In pigs, both IV and IC administration of VEGF/VPF produced a prompt reduction in MAP. Heart rate increased, while cardiac output, stroke volume, left atrial pressure, and total peripheral resistance all declined to a similar, statistically significant degree in both IV and IC groups. Epicardial echocardiography disclosed neither global nor segmental wall motion abnormalities in response to VEGF/VPF. We conclude that (1) VEGF/VPF-stimulated release of NO, previously suggested in vitro, occurs in vivo; (2) this finding suggests that functional VEGF/VPF receptors are present on quiescent adult endothelium, consistent with a maintenance function for VEGF/VPF, which may include regulation of NO; and (3) the preserved response of HC rabbits suggests that endothelial cell receptors for VEGF/VPF are spared in the setting of hypercholesterolemia. PMID- 9409259 TI - Activation of mitogen-activated protein kinases (ERK/JNK) and AP-1 transcription factor in rat carotid arteries after balloon injury. AB - Smooth muscle cell proliferation is a key event in neointimal formation after balloon angioplasty. The molecular signals that mediate this process have yet to be identified. Mitogen-activated protein (MAP) kinases are thought to play a pivotal role in transmitting transmembrane signals required for cell proliferation in vitro. The present studies were designed to investigate whether the signal transduction pathways of MAP kinases were involved in the development of restenosis in the injured arteries. Rat carotid arteries were isolated at various time points after balloon injury, and activities of MAP kinases, including extracellular signal-regulated kinases (ERK), and stress activated protein kinases (SAPK)/c-Jun N-terminal protein kinases (JNK), were determined in protein extracts of the vasculature using protein kinase assay and Western blot analysis. After balloon angioplasty, ERK2 and JNK1 activities in the vessel wall increased rapidly, reached a high level in 5 minutes and maintained for 1 hour. A sustained increase in ERK2 kinase activity was observed over the next 7 days in the arterial wall and 14 days in neointima after injury. In contrast, opposite and uninjured arteries did not show significant changes in these kinase activities. Concomitantly, Western blot analysis confirmed that the ERK2 kinase in the injured vessels was indeed activated or phosphorylated, showing a slowly migrating species of a 42-kDa protein containing phosphorylated tyrosine. Kinase activation is followed by an increase in c-fos and c-jun gene expression and enhanced activator protein 1 (AP-1) DNA-binding activity. Thus, balloon injury rapidly activates the MAP kinases in rat carotid arteries. These kinase activations may be crucial in mediating smooth muscle cell proliferation in response to vascular angioplasty. PMID- 9409260 TI - Genetic factors precipitating type III hyperlipoproteinemia in hypolipidemic transgenic mice expressing human apolipoprotein E2. AB - Several factors are hypothesized to precipitate or exacerbate type III hyperlipoproteinemia (HLP) in humans. Among such factors are those that directly overload remnant lipoprotein production or disrupt removal pathways, including an increased ratio of apolipoprotein (apo) E2 to normal apoE, overproduction of apoB containing lipoproteins, and decreased LDL receptor activity. Hypolipidemic apoE2 transgenic mice bred onto an apoE-null background had dramatically higher plasma total cholesterol (192 +/- 26 mg/dL for males, 203 +/- 40 mg/dL for females) and triglyceride (295 +/- 51 mg/dL for males, 277 +/- 58 mg/dL for females) levels than apoE2 mice with endogenous mouse apoE. Thus, eliminating normal apoE in the presence of apoE2 (thereby increasing the relative abundance of the defective ligand) can convert a hypolipidemic to a hyperlipidemic phenotype. Hypolipidemic apoE2 transgenic mice overexpressing human apoB had moderate remnant accumulation compared with apoE2-only or apoB-only transgenic mice, indicating that overproduction of apoB-containing lipoproteins in the presence of apoE2 can augment remnant production. Hypolipidemic apoE2 transgenic mice bred-onto an LDL receptor-null background had markedly higher plasma total cholesterol (288 +/- 51 mg/dL for males, 298 +/- 73 mg/dL for females) and triglyceride (356 +/- 72 mg/dL for males, 317 +/- 88 mg/dL for females) levels than apoE2-only mice, and remnant accumulation increased even in apoE2 mice with a heterozygous LDL receptor knockout background (compared with apoE2-only mice), suggesting that reducing or eliminating a major receptor-mediated remnant-removal pathway in the presence of apoE2 can also precipitate a hyperlipidemic phenotype. In all cases where either lipoprotein remnant production or removal pathways were severely stressed, increased remnant accumulation was apparent. As judged by the chemical characteristics of the remnant lipoproteins, the lipoprotein phenotype was quite similar to that of human type III HLP, especially in the apoE2-expressing mice with no endogenous apoE or LDL receptors, and thus these mice represent improved models of the disorder. PMID- 9409261 TI - The Arg353Gln polymorphism reduces the level of coagulation factor VII. In vivo and in vitro studies. AB - Factor VII levels are regulated by environmental and genetic factors. Two polymorphisms, a G-to-A transversion at nucleotide 10,976 resulting in Arg353Gln and a decanucleotide insert at position -323 in the 5'-flanking region of the factor VII gene, have been associated with a 20% to 25% reduction in plasma factor VII levels. However Arg353Gln almost always segregates on alleles containing the insert in UK and Italian populations, thereby making it impossible to independently evaluate the impact of Arg353Gln on factor VII levels in these ethnic groups. We have evaluated the influence of genotype on factor VII levels in 99 healthy Polish blood donors and observed that Arg353Gln frequently occurs in the absence of the insert. In univariate analysis, the mean levels of factor VII coagulant activity (VII:C) and factor VII antigen (VII:Ag) were significantly lower in 16 people who were heterozygous for Arg353Gln and the insert compared with 72 normal subjects who had neither Arg353Gln nor the insert (88.8% of normal and 83.1% versus 102% and 100%, P = .019 and P = .0003, respectively). In nine subjects heterozygous for Arg353Gln alone, VII:C and VII:Ag were significantly decreased compared with the normal subjects (81.9% and 83%, respectively, P = .007 and P = .004). In multivariate analysis, Arg353Gln but not the insert significantly reduced VII:C and VII:Ag after adjustment for age and plasma triglycerides (P < .05 and P = .02, respectively). To evaluate the mechanism responsible for reduced factor VII levels in individuals with Arg353Gln, we performed transient transfection assays with factor VII cDNA containing the base substitution resulting in Gln353 and wild-type factor VII cDNA in COS-1 cells. The levels of VII:Ag in the cell lysates were similar, but the amino acid substitution significantly reduced factor VII secretion into the media to 74.9% of wild-type (P = .0001). Based on these in vivo and in vitro studies, we conclude that the Arg353Gln polymorphism alone can decrease plasma factor VII levels. PMID- 9409262 TI - In vivo glucosylated LpA-I subfraction. Evidence for structural and functional alterations. AB - This study compared the structural and functional properties of glucosylated and non-glucosylated LpA-I particle subfractions (GLpA-I and NGLpA-I, respectively) isolated from patients with poorly controlled type 1 (insulin-dependent) diabetes. Compared with NGLpA-I, GLpA-I showed an enrichment in triglycerides (P < .05) and a depletion in phospholipid (P < .05) content. Moreover, the triglycerides-to-cholesteryl esters ratio was increased (P < .05), suggesting an increased cholesteryl ester transfer protein activity and a possible transport defect that accelerates atherogenesis. The surface-to-core constituents ratio, an indirect estimate of particles size, is lower in GLpA-I (P < .01) than in NGLpA I, correlating well with a larger median size (P < .05) as seen by electron microscopy. The apolipoprotein (apo) A-I conformation was evaluated through determination of the immunological accessibility of three different domains defining specific epitopes for anti-apo A-I monoclonal antibodies. We observed a marked decreased accessibility for two of these regions, which interestingly have already been implicated in the interaction with cells. Cell culture data suggest that nonenzymatic glycosylation occurring on apo A-I can modify lipoprotein function, since it results in a decreased binding of GLpA-I to HeLa cells and impaired cholesterol efflux from Fu5AH rat hepatoma cells. PMID- 9409263 TI - Common C-to-T substitution at position -480 of the hepatic lipase promoter associated with a lowered lipase activity in coronary artery disease patients. AB - We studied the molecular basis of low hepatic lipase (HL) activity in normolipidemic male patients with angiographically documented coronary artery disease (CAD). In 18 subjects with a lowered HL activity (< 225 mU/mL), all nine exons of the HL gene and part of the promoter region (nucleotides -524 to +7) were sequenced. No structural mutations in the coding part of the HL gene were found, but 50% of the subjects showed a C-to-T substitution at nucleotide -480. Screening for the base substitution in 782 patients yielded an allele frequency of 0.213 (297 heterozygotes, 18 homozygotes). In a group of 316 nonsymptomatic control subjects, the allele frequency was 0.189, which is significantly less than in the CAD patients (P = .035). In the CAD patients, the C-to-T substitution was associated with a lowered lipase activity (heterozygotes -15%, homozygotes 20%). The patients were divided into quartiles on the basis of HL activity. Sixty percent (allele frequency 0.32) of the patients in the lowest quartile (HL activity < 306 mU/mL) had the gene variant against 27% (allele frequency 0.14) in the highest quartile (HL activity > 466 mU/mL). In the noncarriers, but not in the carriers, HL activity was related with plasma insulin, being increased at higher insulin concentration. Homozygous carriers had a significantly higher HDL cholesterol level-than noncarriers (1.13 +/- 0.28 mmol/L versus 0.92 +/- 0.22 mmol/L, P < .02). Our results show that a C-to-T substitution at -480 of the HL promoter is associated with a lowered HL activity. The base substitution, or a closely linked gene variation, may contribute to the variation in HL activity and affect plasma lipoprotein metabolism. PMID- 9409264 TI - Elevated circulating levels of inflammatory cytokines in patients with abdominal aortic aneurysm. AB - The basic feature in the pathogenesis of abdominal aortic aneurysm (AAA) is the degradation of extracellular matrix components. This process is induced partly by cytokines secreted from inflammatory and mesenchymal cells. Circulating levels of inflammatory cytokines were studied in AAA patients and compared with subjects suffering from atherosclerotic disease only. Furthermore, the predictive value of cytokine concentrations was evaluated for aneurysm expansion rate. Circulating levels of interleukin 1 beta (IL-1 beta), interleukin 6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), and interferon-gamma (IFN-gamma) were measured in 50 AAA patients (40 men, 10 women), 42 patients with coronary heart disease (CHD) (23 men, 19 women), and 38 controls whose angiogram was normal (17 men, 21 women). No differences in cytokine concentrations were found between the CHD patients and the controls. AAA disease was found to be associated with significantly higher IL-1 beta and IL-6 concentrations in both male patients (median concentrations of 19.40 pmol/L and 6.45 pmol/L, respectively) and female patients (19.26 pmol/L and 7.99 pmol/L) than in either the CHD patients or the controls (P < .005). TNF-alpha levels were slightly higher in the AAA patients (1.64 pmol/L in the males and 1.59 pmol/L in the females) than in the other groups (P < .05). IFN-gamma levels were elevated significantly in the female AAA patients (3.75 pmol/L) compared with levels found in the other female (P < .05) or male (P < .01) patient groups. The measured cytokine concentrations were not related to the size of the aneurysm or the maximal thickness of the thrombus within the aneurysm. IFN-gamma concentration showed a significant positive correlation to the aneurysm expansion (R = .37, P < .02) and negative correlation to the concentration of aminoterminal propeptide of type III procollagen during 6 month follow up (R = -.42, P < .005). The results show that circulating levels of inflammatory cytokines are elevated in patients with AAA disease, suggesting that the production of these cytokines is increased in these patients compared with CHD patients and controls. Elevated INF-gamma concentrations seem to predict an increased rate of expansion in AAA. PMID- 9409265 TI - Induction of vascular SMC proliferation by urokinase indicates a novel mechanism of action in vasoproliferative disorders. AB - The urokinase-type plasminogen activator (UPA) and its receptor are expressed in the vasculature and are involved in cell migration and remodeling of the extracellular matrix in the neointima. Vessels with atherosclerosis or neointimal hyperplasia, when compared with normal vessels, contain high UPA activity as well as increased levels of UPA receptor. In this study, we have identified the stimulation of vascular smooth muscle cell proliferation as a novel activity for UPA in the vessel wall. High-molecular-weight-UPA (12-200 nmol/L range) stimulated DNA synthesis and cell proliferation, which was half that induced by fetal calf serum or by platelet-derived growth factor-BB. UPA did not induce growth of endothelial cells, and tissue-type plasminogen activator showed no activity on either cell type. Induction of proliferation required the complete UPA molecule but was independent of the proteolytic activity of UPA, whereas neither the amino-terminal fragment nor the catalytic domain by itself was mitogenic. UPA also stimulated c-fos/c-myc mRNA expression and mitogen-activated protein kinase activity in smooth muscle cells. Blocking monoclonal antibodies against the UPA receptor and the enzymatic removal of receptors were ineffective in inhibiting the mitogenic effect of UPA, suggesting a UPA receptor-independent mechanism. Thus, we provide evidence for a novel function of UPA on vascular smooth muscle cell proliferation that, together with its previously documented involvement in regulating pericellular proteolysis-related events and cell migration, provides additional evidence for a role in the pathogenesis of atherosclerosis/restenosis. PMID- 9409266 TI - Paracetamol catalyzes myeloperoxidase-initiated lipid oxidation in LDL. AB - The oxidative modification of LDL may play a significant role in atherogenesis. Myeloperoxidase (MPO) expressed in human atherosclerotic plaques has been suggested to be operative in vivo, making LDL atherogenic. Tyrosyl radicals generated by MPO have been shown to act as physiological pro-oxidants of lipid peroxidation in LDL. Assuming that a variety of phenolic compounds are able to form phenoxyl radicals when exposed to peroxidases, we tested the ability of paracetamol, a known analgesic drug with a tyrosine-like monophenolic structure, to act as a pro-oxidant of lipid peroxidation in LDL. Spectroscopic analyses indicated that paracetamol, similar to tyrosine, could undergo peroxidase-induced phenoxyl radical formation, which was inhibited by the radical scavenger ascorbic acid as well as by heme poisons and catalase. Measurement of conjugated dienes and lipid hydroperoxides in LDL preparations exposed to MPO/H2O2 in the absence or presence of paracetamol revealed that the drug could act as a catalyst of lipid oxidation in LDL. Similar results were found when LDL oxidation was performed with activated human neutrophils, which use MPO to promote lipid peroxidation. In conclusion, the results suggest that paracetamol could act, via a phenoxyl radical, as a catalyst of LDL oxidative modification by MPO. PMID- 9409267 TI - Evidence against an effect of endothelin-1 on blood coagulation, fibrinolysis, and endothelial cell integrity in healthy men. AB - On the basis of an array of preclinical experimental results, it has been widely assumed that endothelin-1 (ET-1) may affect blood coagulation, fibrinolysis, and endothelial cell function, thereby playing a pathophysiological role in various cardiovascular diseases in humans. However, confirmation of this assumption is still lacking. ET-1 or placebo was administered intravenously to 12 healthy volunteers in a prospective, randomized, double-blind, crossover trial. Pathophysiologically relevant concentrations of ET-1 (an approximate threefold increase of normal blood levels) causing hemodynamic effects were reached by continuous intravenous infusion for 6 hours. Components of the coagulation (thrombin-antithrombin complexes, prothrombin fragment F1 + 2, activated factor VII, and factor VII antigen) and fibrinolytic (fibrin split product D-dimer, plasmin-plasmin inhibitor complex, tissue-type plasminogen activator, urokinase type plasminogen activator, and plasminogen activator inhibitor-1) systems and markers of endothelial cell perturbation/dysfunction (von Willebrand factor and thrombomodulin) were measured before the start of infusion and after 2, 6, 12, and 24 hours. Comparing changes in the plasma concentrations of these parameters during and after infusion of ET-1 and placebo, we found no specific effects of ET 1. In contrast to previous reports from preclinical experiments, ET-1 does not appear to affect coagulation or fibrinolysis, nor does this peptide induce relevant endothelial cell perturbations in humans. PMID- 9409268 TI - Genistein, the dietary-derived angiogenesis inhibitor, prevents LDL oxidation and protects endothelial cells from damage by atherogenic LDL. AB - There is now growing evidence that the oxidative modification of LDL plays a potential role in atherosclerosis. In this study, genistein, a compound derived from a soy diet with a flavonoid chemical structure (4',5,7 trihydroxyisoflavone), which was found to inhibit angiogenesis, has been evaluated for its ability to act as an LDL antioxidant and a vascular cell protective agent against oxidized LDL. The results showed that genistein was able to inhibit the oxidation of LDL in the presence of copper ions or superoxide/nitric oxide radicals as measured by thiobarbituric acid-reactive substance formation, alteration in electrophoretic mobility, and lipid hydroperoxides. Bovine aortic endothelial cell- and human endothelial cell mediated LDL oxidation was also inhibited in the presence of genistein. The 7-O glucoside of genistein, genistin, was much less effective in inhibiting LDL oxidation in the cell-free and cell-mediated lipoprotein-oxidating systems. Incubating human endothelial cells in the absence or presence of genistein and challenging the cells with already oxidized lipoprotein revealed that in addition to its antioxidative potential during LDL oxidating processes, genistein effectively protected the vascular cells from damage by oxidized lipoproteins. The tyrosine kinase inhibitor genistein was found to block upregulation of two tyrosine-phosphorylated proteins of 132 and 69 kDa in endothelial cells induced by oxidized LDL. Parallel experiments with the inactive analogue daidzein, however, showed that the cytoprotective effect of the isoflavones seems not to be dependent on tyrosine phosphorylation. Our findings will support the suggested and documented beneficial action of a soy diet in preventing chronic vascular diseases and early atherogenic events. PMID- 9409269 TI - A prothrombin gene mutation is significantly associated with venous thrombosis. AB - This case-control study examined the prevalence of a prothrombin gene mutation in the 3'-untranslated region (UTR) first reported by Poort et al in Dutch subjects with a history of venous thrombosis and in matched control subjects without a history of thrombosis. We tested the hypothesis that the presence of the 3'UTR prothrombin mutation would convey a higher risk of venous or arterial thrombosis and therefore would be found in a higher-than-normal percentage of subjects with a history of thrombosis. Our study included 100 subjects: 50 with a history of thrombosis (21 with venous thrombosis and 29 with arterial thrombosis, who had been recruited from an anticoagulation clinic) and 50 control subjects without a history of thrombosis. DNA from these subjects was analyzed by polymerase chain reaction and agarose gel electrophoresis. We found a statistically significant increase in the prevalence of the 3'UTR mutation in subjects with a history of venous thrombosis compared with subjects without thrombosis. The prevalence of the 3'UTR prothrombin mutation was 19% (4/21;3 heterozygous and 1 homozygous) in subjects with a history of venous thrombosis, 0% (0/29) in subjects with a history of arterial thrombosis, and 2% (1/50) in control subjects (P < .0245, by Fisher's exact test for comparison of subjects with versus those without a history of venous thrombosis). The G-->A mutation at nucleotide 20,210 in the 3'UTR was confirmed by direct DNA sequencing. The similar increased prevalence of the 3'UTR mutation in subjects with venous thrombosis in our population and in the Dutch population studied by Poort et al suggests that this mutation is an important risk factor for venous thrombosis in the general white population. PMID- 9409270 TI - The apolipoprotein E and beta-fibrinogen G/A-455 gene polymorphisms are associated with ischemic stroke involving large-vessel disease. AB - The relationship between the apolipoprotein E (apoE) and beta-fibrinogen G/A-455 polymorphisms and cerebrovascular disease (CVD) was examined in the present study. We compared 227 patients with the subtypes of CVD (large-vessel disease, lacunar stroke, cardiac embolism, or undetermined pathomechanisms) with 225 control subjects. The occurrence of apoE isoforms (E2, E3, and E4) and the beta fibrinogen G/A-455 genotype was determined in these individuals. No differences in apoE polymorphisms or allele frequencies between the CVD patients and control subjects were found. However, analysis of apoE genotypes as a function of stroke subtype revealed that the apoE4 allele was significantly more common in those patients with macroangiopathy-associated CVD. The only CVD risk factor that distinguished patients with the E4 allele from those with other apoE genotypes was elevated cholesterol. No association between the beta-fibrinogen G/A-455 polymorphism and CVD was found. However, homozygosity for the A allele was more common in patients with CVD resulting from large-vessel disease. These data demonstrate that the apoE4 allele and the AA genotype of the beta-fibrinogen G/A 455 polymorphism occur significantly more frequently in patients with CVD resulting from stenosis of large, brain-supplying vessels. Such genetic analyses may further our understanding of the etiology of cerebrovascular disease. PMID- 9409271 TI - Effect of advanced glycation end product-modified albumin on tissue factor expression by monocytes. Role of oxidant stress and protein tyrosine kinase activation. AB - Diabetes is associated with a hypercoagulable state that contributes to macrovascular complications, including cardiovascular events. The glycation reaction, a consequence of chronic hyperglycemia, has also been implicated in the pathogenesis of diabetic complications. Glycated proteins have receptors on monocytes and generate reactive oxygen species that can regulate the expression of a number of genes. As abnormal monocyte expression of tissue factor (TF), the main initiator of the coagulation cascade, is responsible for thrombosis in a number of clinical settings, we studied the effect of glycated albumin on monocyte TF expression. Mononuclear cells were incubated with glycated albumin for 24 hours, and monocyte TF activity was measured with a plasma recalcification time assay; TF antigen was measured by ELISA and TF mRNA by RT-PCR. Glycated albumin induced blood monocyte expression of the procoagulant protein TF at the mRNA level. Oxidative stress appeared to be involved in this effect, as the antioxidant N-acetylcysteine diminished TF mRNA accumulation in stimulated monocytes. Hydroxyl radicals, which may be generated inside cells from H2O2 via the Fenton reaction, also appeared to be involved in this effect, as hydroxyl radical scavengers downregulated TF activity and antigen levels (but not TF mRNA). Finally, the involvement of activated protein tyrosine kinase in the transmission of the signal from the membrane to the nucleus was suggested by the inhibitory effect of herbimycin A. These results point to a new mechanism for the hypercoagulability often described in diabetic patients and suggest that antioxidants or protein tyrosine kinase inhibitors might be of therapeutic value in this setting. PMID- 9409272 TI - Evidence for the migration of rat aortic endothelial cells toward the heart. AB - Most vascular endothelial cells at the edge of experimentally induced wounds have their centrosomes oriented toward the wound in the direction of cell migration. The finding that the centrosomes in endothelial cells of non-wounded aorta and vena cava are also oriented toward the heart suggested the hypothesis that endothelial cells are normally migrating in this direction. To test this hypothesis, endothelial cells in a segment of the rat abdominal aorta were labeled with a relatively nontoxic dye, 1,1'-dioctadecyl-3,3,3',3' tetramethylindocarbocyanine perchlorate (DiI), and the position of the labeled cells was determined 3 and 6 weeks later. The results obtained showed that in 6 of the 9 rat aortas examined at 3 weeks and 15 of the 20 rat aortas examined at 6 weeks, DiI-labeled endothelial cells had migrated various distances up to 5000 microns toward the heart. In contrast, no migration of endothelial cells was detected at the opposite end of the labeled segment, in the direction away from the heart. These results demonstrate that vascular endothelial cells in the abdominal aorta of the rat are not stationary but are migrating toward the heart. The significance of the migration of endothelial cells toward the heart is presently unknown; however, it would be interesting to explore whether or not the impairment of this migration may contribute to disease processes in which the ability to maintain an intact and normally functioning endothelial cell lining is compromised as in atherosclerosis. PMID- 9409273 TI - Effect of phenotype on the transcription of the genes for platelet-derived growth factor (PDGF) isoforms in human smooth muscle cells, monocyte-derived macrophages, and endothelial cells in vitro. AB - Proliferation of arterial smooth muscle cells (ASMCs) contributes considerably to enlargement of the arterial wall during atherosclerosis. The platelet-derived growth factor (PDGF) is a well-known mitogen and chemoattractant for ASMCs. Quantitative reverse transcription-polymerase chain reaction showed that cells appearing in atherosclerotic lesions, such as ASMCs, endothelial cells, and monocytes/macrophages, expressed mRNAs for both PDGF A and B chains in vitro, with the highest expression in endothelial cells. On proliferation, ASMCs and endothelial cells upregulated PDGF A mRNA. Differentiation of macrophages increased the amount of both mRNAs. Thus, the regulation of PDGF A- and B-chain expression depends on cell types and phenotypic states of the cells, which have also been found in vivo in human atherosclerotic lesions. PDGF A can be produced as short and long isoforms. The latter binds with high affinity to glycosaminoglycans. Irrespective of phenotype, only the minor part of total PDGF A mRNA consisted of the long variant in ASMCs, while endothelial cells produced 40% of total PDGF A as the long form. The differentiation of macrophages increased the production of the long PDGF A mRNA from 10% to 40%. Thus, increasing numbers of stimulated cells in the atherosclerotic lesion may increase the transcription of PDGF isoforms, and particularly of the long PDGF A isoform. Together with increasing amounts of ASMC-derived proteoglycans in developing lesions, this may contribute to accumulation of PDGF in the arterial wall matrix, resulting in prolonged stimulation of ASMCs. PMID- 9409274 TI - Effects of dietary fat quality and quantity on postprandial activation of blood coagulation factor VII. AB - Acute elevation of the coagulant activity of blood coagulation factor VII (FVIIc) is observed after consumption of high-fat meals. This elevation is caused by an increase in the concentration of activated FVII (FVIIa). In a randomized crossover study, we investigated whether saturated, monounsaturated, or polyunsaturated fats differed regarding postprandial activation of FVII. Eighteen healthy young men participated in the study. On 6 separate days each participant consumed two meals (times, 0 and 1 3/4 hours) enriched with 70 g (15 and 55 g) of either rapeseed oil, olive oil, sunflower oil, palm oil, or butter (42% of energy from fat) or isoenergetic low-fat meals (6% of energy from fat). Fasting and series of nonfasting blood samples (the last at time 8 1/2 hours) were collected. Plasma triglycerides, FVIIc, FVIIa, and free fatty acids were analyzed. There were marked effects of the fat quantity on postprandial responses of plasma triglycerides, FVII, and free fatty acids. The high-fat meals caused, in contrast to the low-fat meals, considerable increases in plasma triglycerides. Plasma levels of FVIIc and FVIIa peaks were 7% and 60% higher after consumption of high fat meals than after consumption of low-fat meals. The five different fat qualities caused similar postprandial increases in plasma triglycerides, FVIIc, and FVIIa. These findings indicate that high-fat meals may be prothrombotic, irrespective of their fatty acid composition. The postprandial FVII activation was not associated with the plasma triglyceride or free fatty acid responses. PMID- 9409275 TI - Chronic Chlamydia pneumoniae infection is associated with a serum lipid profile known to be a risk factor for atherosclerosis. AB - Chlamydia pneumoniae infection has been associated with coronary heart disease. To evaluate the mechanisms of this association, we studied whether chronic C. pneumoniae infection affects serum lipid values similarly to acute infections. Triglyceride, total and HDL cholesterol concentrations, and C. pneumoniae antibodies were measured from paired serum samples of 415 Finnish males taken 3 years apart. Chronic infection, defined as persistent IgG and IgA antibodies, was found in 20%, and the antibodies were negative (IgG < 32 and IgA < 16 in both samples) in 15% of the cases studied. The serum triglyceride and total cholesterol concentrations were higher in the subjects with a chronic C. pneumoniae infection than in the subjects with no antibodies (1.23 versus 1.03 mmol/L and 6.41 versus 6.31 mmol/L, respectively). The HDL cholesterol concentrations and the ratios of HDL cholesterol to total cholesterol were significantly decreased in the subjects with chronic infection (1.24 versus 1.36 mmol/L, P = .026; and 0.19 versus 0.22, P = .018, respectively). Chronic C. pneumoniae infection seems to be associated with a serum lipid profile considered to increase the risk of atherosclerosis. This finding supports the hypothesis that infections play a role in the pathogenesis of atherosclerosis. PMID- 9409276 TI - ApoE genotype does not predict lipid response to changes in dietary saturated fatty acids in a heterogeneous normolipidemic population. The DELTA Research Group. Dietary Effects on Lipoproteins and Thrombogenic Activity. AB - Recent studies have suggested that variations in apoE genotypes may influence the magnitude of plasma lipid changes in response to dietary interventions. We examined the ability of apoE genotype to predict plasma lipid response to reductions in percent of calories from total fat (TF) and saturated fat (SF) in a normolipidemic study population (n = 103) heterogeneous with respect to age, gender, race, and menopausal status. Three diets, an average American diet (34.3% TF, 15.0% SF), an AHA Step 1 diet (28.6% TF, 9.0% SF), and a low saturated fat (Low-Sat) diet (25.3% TF, 6.1% SF) were each fed for a period of 8 weeks in a three-way crossover design. Cholesterol was kept constant at 275 mg/d; monounsaturated and polyunsaturated fat were kept constant at approximately 13% and 6.5% of calories, respectively. Fasting lipid levels were measured during each of the final 4 weeks of each diet period. Participants were grouped by apoE genotype: E2 (E2/2, E2/3, E2/4); E3 (E3/3); E4 (E3/4, E4/4). Relative to the average American diet, both the Step 1 and Low-Sat diets significantly reduced total cholesterol, LDL cholesterol, and HDL cholesterol in all three apoE genotype groups. No evidence of a significant diet by genotype interaction, however, could be identified for any of the measured lipid and lipoprotein end points. Additional analysis of the data within individual population subgroup (men and women, blacks and whites) likewise provided no evidence of a significant diet by genotype interaction. Thus, in a heterogeneous, normolipidemic study population, apoE genotype does not predict the magnitude of lipid response to reductions in dietary saturated fat. PMID- 9409277 TI - Hyperhomocyst(e)inemia and a common methylenetetrahydrofolate reductase mutation (Ala223Val MTHFR) in patients with inherited thrombophilic coagulation defects. AB - To assess whether certain abnormalities of the sulfated amino acid metabolism are associated with the occurrence of thromboembolic events in patients with inherited thrombophilic conditions, the levels of homocyst(e)ine, before or after methionine load, and the presence of the Ala223Val substitution in the 5,10 methylenetetrahydrofolate reductase (MTHFR) were evaluated in 119 subjects with a congenital single thrombophilic condition (type I deficiency of antithrombin n = 10, protein C n = 24, protein S n = 16; activated protein C resistance due to factor V Leiden mutation n = 69). Sixty-three subjects had experienced at least one documented thrombotic event, while the remaining 56 subjects were still free from any thrombotic symptom. Our results show that (1) high homocyst(e)ine levels, either in fasting condition or after methionine load, were not more frequent in subjects with inherited thrombophilic alterations (14.4%) than in normal control subjects (10% by definition) and (2) the frequency of hyperhomocyst(e)inemia was similar in thrombophilic subjects, who already have (14.3%) or have not (14.6%) experienced thrombotic events. As regards the MTHFR mutation, the homozygous condition was present in 23.2% of the thrombophilic patients versus 17.5% in the control subjects, a nonsignificant difference. The mutation was slightly more frequent in those thrombophilic subjects who had suffered a thrombotic episode (25.5%) versus those with no thrombosis (20.8%), with odds ratios of 1.61 (confidence interval (CI) = 0.58-4.52) and 1.24 (CI = 0.42-3.43), respectively. These differences were also nonsignificant. It is concluded that in subjects with inherited thrombophilias, a condition of hyperhomocyst(e)inemia "per se" is not a factor increasing the risk of thrombosis. The risk enhancement conferred by the MTHFR mutation, if any, seems to be slight or limited, and its significance could be ascertained only in a large multicenter trial. PMID- 9409278 TI - Consequences of hyperhomocyst(e)inemia on vascular function in atherosclerotic monkeys. AB - Moderate elevation of plasma homocyst(e)ine is associated with increased risk for atherosclerotic vascular disease. In a previous study, we observed impaired vascular function in nonatherosclerotic monkeys with moderate hyperhomocyst(e)inemia. In this study, we tested the hypothesis that dietary intervention to lower plasma homocyst(e)ine corrects vascular dysfunction in atherosclerotic monkeys. Cynomolgus monkeys were fed an atherogenic diet that produces both hypercholesterolemia and moderate hyperhomocyst(e)inemia. After 17 months, the atherogenic diet was supplemented with B vitamins (5 mg folic acid, 400 micrograms vitamin B-12, and 20 mg vitamin B-6 daily) for 6 months. Total plasma homocyst(e)ine decreased from 12.8 +/- 2.8 to 3.5 +/- 0.3 mumol/L (n = 9; mean +/- SE; P < .01) after vitamins were added to the diet, but plasma cholesterol remained elevated (522 +/- 63 versus 514 +/- 41 mg/dL; P > .05). In response to intra-arterial infusion of collagen, blood flow to the leg decreased by 30 +/- 3% and 38 +/- 5%, respectively, before and after vitamin supplementation (P > .05). In vivo responses of resistance vessels to endothelium dependent vasodilators (acetylcholine or ADP) were impaired at baseline and did not improve after vitamin supplementation. In carotid artery studied ex vivo, relaxation to low doses of acetylcholine improved after vitamin supplementation, but maximal relaxation remained impaired. Ex vivo thrombomodulin anticoagulant activity was threefold higher in monkeys fed the atherogenic diet (with or without B vitamins) than in normal monkeys (P < .05). We conclude that normalization of plasma homocyst(e)ine is insufficient to restore normal vascular function in atherosclerotic monkeys with persistent hypercholesterolemia and that atherosclerosis, with or without hyperhomocyst(e)inemia, is associated with elevated thrombomodulin activity. PMID- 9409279 TI - Two alleles of the human paraoxonase gene produce different amounts of mRNA. An explanation for differences in serum concentrations of paraoxonase associated with the (Leu-Met54) polymorphism. AB - In a recent study we demonstrated that a polymorphism of the human paraoxonase gene affecting position 54 was linked to variations in serum concentrations of the enzyme. L allele carriers (leucine at position 54) have significantly higher concentrations of paraoxonase than M allele carriers (methionine at position 54). In the present study we examined the hypothesis that differences in mRNA production could contribute to variations in serum concentrations. Relative concentrations of L and M type mRNA were analyzed in total RNA extracted from heterozygous liver samples. This was achieved by cDNA synthesis, polymerase chain reaction amplification of the cDNA fragment containing the 54 polymorphism and restriction analysis to identify radiolabeled end fragments of L and M alleles. An allele mixing experiment using total RNA from liver samples of LL and MM homozygotes demonstrated the sensitivity of the approach to changes in the relative concentrations of each type of RNA. In 8 of 10 heterozygous samples, an excess of L allele type mRNA was observed. Overall there was a significantly higher level of L type mRNA (L:M ratio of 2.51 +/- 1.41, n = 10, P < .01). These results support our hypothesis that increased concentrations of serum paraoxonase arise from greater production of L allele mRNA. In two samples, the L:M ratio was close to or below 1.0. This is consistent with the known spectrum of paraoxonase serum concentrations associated with the L and M alleles and suggests that factor(s) that preferentially modulate allele expression are usually, but not uniformly, associated with the L allele. PMID- 9409280 TI - Altered transfer of cholesteryl esters and phospholipids in plasma from alcohol abusers. AB - The net mass transfer (NMT) of cholesteryl esters (CEs), triglycerides (TGs), and phospholipids (PLs) between lipoproteins was measured after incubation of fresh plasma for up to 2 hours from 18 male alcohol abusers and 17 male volunteer control subjects. In alcohol abusers the mean value of CE NMT was 3.7 nmol.mL-1.h 1 from apolipoprotein B-containing lipoproteins (apoB-containing lipoproteins) to HDL and in control subjects 8.7 nmol.mL-1.h-1 from HDL to apoB-containing lipoproteins. The NMT of PL was higher in alcohol abusers than in control subjects (35.0 vs 11.6 nmol.mL-1.h-1 from apoB-containing lipoproteins to HDL, respectively), and plasma PL transfer protein (TP) activity was 33% higher (P < .05) in alcohol abusers than in control subjects. The lack of correlation between the NMTs and CETP and PLTP activities suggests that the NMT could more closely reflect the role of lipoprotein properties in reverse cholesterol transport in vivo, whereas in vitro activities reflect the total capacity of transfer but not its direction. The rate of CE NMT from HDL to apoB-containing lipoproteins was dependent on the VLDL TG concentration. Moreover, at low VLDL TG levels, the increased HDL cholesterol concentration in alcohol abusers reversed the direction of CE NMT. This situation could be reconstructed in the plasma of control subjects by adding autologous HDL or VLDL to mimic the lipoprotein profiles of the alcohol abusers. Addition of VLDL enhanced the CE NMT from HDL to apoB containing lipoproteins, whereas addition of HDL had an opposite effect, and at higher HDL levels, even reversed the direction of CE NMT. In conclusion, the NMT of CE and PL in alcohol abusers differs from that in control subjects. The concentrations of HDL and VLDL seem to be the major determinants of the direction of CE NMT in alcohol abusers. PMID- 9409281 TI - Liver LDL receptor mRNA expression is decreased in human ApoB/CETP double transgenic mice and is regulated by diet as well as the cytokine oncostatin M. AB - We have investigated liver LDL receptor mRNA expression in nontransgenic, human cholesteryl ester transfer protein (CETP) transgenic, and human apolipoprotein (Apo) B/CETP double transgenic mice fed a normal chow diet and a high fat, high cholesterol diet (HFHC). Three weeks of HFHC feeding increased total serum cholesterol 1.5-fold in the nontransgenic, 3.1-fold in the CETP transgenic, and 3.4-fold in the ApoB/CETP double transgenic mice. To examine the liver LDL receptor mRNA expression among the different groups of mice fed the normal diet or fed the HFHC diet, we developed a quantitative reverse-transcribed polymerase chain reaction assay in which the LDL receptor mRNA level was normalized with the beta-actin mRNA. The results show that on the normal chow diet, the LDL receptor mRNA expression levels were lower in the ApoB/CETP mice than in the nontransgenic mice and the human CETP transgenic mice. Liver LDL receptor gene expression was lower in all groups of mice fed the HFHC diet, with the lowest level of expression in the ApoB/CETP mice. Similar results were obtained by Northern blot analysis. In addition, we have previously shown that the cytokine oncostatin M (OM) increases LDL receptor gene expression in HepG2 cells. In this study, we used the ApoB/CETP mice as the model system to examine the in vivo activity of OM on liver LDL receptor gene expression. Our data show that OM increased the level of liver LDL receptor mRNA up to 80% to 90% when the animals were fed the HFHC diet. The results from these studies demonstrate that the expression of the liver LDL receptor in the ApoB/CETP mice is suppressed compared with nontransgenic mice and that the expression of the hepatic LDL receptor gene in these mice is subjected to the normal cholesterol feedback regulation. In addition, LDL receptor gene expression in these mice is also inducible by a positive regulator. PMID- 9409282 TI - Assembly and secretion of VLDL in nondifferentiated Caco-2 cells stably transfected with human recombinant ApoB48 cDNA. AB - Intestinal cells secrete apoB48-containing very low density lipoproteins (VLDLs) and chylomicrons for the transport of biliary and dietary lipids. The molecular mechanisms regulating the assembly of intestinal lipoproteins are not known due to a lack of reliable and specific cell culture models. Caco-2 (a human colon carcinoma) cells have been used to study intestinal lipid metabolism. These cells have been shown to secrete both apoB100- and apoB48-containing triglyceride (TG) rich lipoproteins only after differentiation into enterocyte-like cells. To study lipoprotein assembly in nondifferentiated Caco-2 cells, we stably expressed human recombinant apoB48 cDNA under the control of a constitutive cytomegalovirus promoter. Pulse-chase analysis revealed that the majority (> 50%) of apoB48 synthesized was degraded intracellularly in the presence or absence of oleic acid. Transfected nondifferentiated cells secreted lipoproteins with flotation densities similar to those of plasma HDL or LDL when cultured in serum-free or serum-containing media, respectively. Incubation of cells with media containing serum and oleic acid resulted in the secretion of VLDL-like particles. Secretion of VLDL was inhibited (> 80%) by triacsin C due to > 60% inhibition of oleate induced TG synthesis. However, inhibition of cholesteryl ester synthesis by 70% with an acyl coenzyme A:cholesterol acyltransferase inhibitor did not affect VLDL secretion. Efficient assembly of lipoproteins usually requires the microsomal TG transfer protein (MTP). The presence of MTP in transfected Caco-2 cells was investigated by measuring TG transfer activity in microsomal fractions. Microsomal fractions had 0.2% TG transfer activity per hour per microgram of protein, which corresponds to 30% to 60% of the MTP activity present in liver derived cells. To determine whether MTP activity was required for lipoprotein assembly, transfected cells were incubated in the presence of the MTP inhibitor CP-10,447. This compound completely abolished the secretion of apoB. These data show that the transfected cell lines secrete lipoproteins of different densities under different culture conditions and that the assembly of larger VLDL particles requires active TG synthesis and MTP activity. Thus, in nondifferentiated Caco-2 cells, the amount of apoB secreted and not the MTP activity is the limiting factor for lipoprotein assembly. PMID- 9409283 TI - Tissue factor pathway inhibitor in endothelial cells colocalizes with glycolipid microdomains/caveolae. Regulatory mechanism(s) of the anticoagulant properties of the endothelium. AB - Tissue factor pathway inhibitor (TFPI), the main downregulator of the procoagulant activity of tissue factor.factor VIIa complex, locates in human endothelial cells (EC) in culture as well-defined clusters uniformly distributed both on the cell surface and intracellularly. We here demonstrate by immunofluorescence that TFPI colocalizes in EC with caveolin, urokinase-type plasminogen activator receptor, and glycosphingolipids. The localization of TFPI in caveolae in resting endothelium is proved by double immunogold electron microscopy for TFPI and caveolin. After ultracentrifugation of rat lung or EC homogenates through density gradients of Nycodenz, TFPI was highly enriched at densities of 1.05 to 1.08 g/mL, together with caveolin and alkaline phosphatase. By ELISA, more than half of the cellular TFPI was detected in Triton X-100 insoluble extracts of EC. TFPI incorporates [1-3H]ethanolamine and is cleaved from the cell surface by phosphatidylinositol-phospholipase C, indicating a specific glycosylphosphatidylinositol-anchorage mechanism for TFPI in the plasma membrane. Clustering of TFPI and its localization in caveolae are dependent on the presence of cholesterol in the membrane. Agonist-induced stimulation of EC caused marked changes of distribution for both TFPI and caveolin at subcellular level, with subsequent increase of the cell surface-associated inhibitory activity toward tissue factor.factor VIIa. Our findings suggest that, beside their function in transcytosis, potocytosis, cell surface proteolysis, and regulation of signal transduction, caveolae also play a direct role in the regulation of EC anticoagulant properties. PMID- 9409284 TI - Vascular superoxide dismutase deficiency impairs endothelial vasodilator function through direct inactivation of nitric oxide and increased lipid peroxidation. AB - Nitric oxide (NO) and superoxide are both constitutive products of the endothelium. Because NO is readily inactivated by superoxide, the bioactivity of endothelium-derived NO (EDNO) is dependent on local activity of superoxide dismutase (SOD). We examined the effects of chronic inhibition of copper-zinc SOD (CuZnSOD) using a rat model of dietary copper restriction. Male weanling Sprague Dawley rats were fed a Cu-deficient diet and received either no Cu replacement (Cu-deficient) or Cu in the drinking water (Cu-sufficient). Compared with Cu sufficient animals, Cu-deficiency was associated with a 68% reduction in CuZnSOD activity and a 58% increase in vascular superoxide as estimated by lucigenin chemiluminescence (both P < .05). Compared with Cu-sufficient animals, arterial relaxation in the thoracic aorta from Cu-deficient animals was 10-fold less sensitive to acetylcholine, a receptor-dependent EDNO agonist, but only 1.5-fold less sensitive to A23187, a receptor-independent EDNO agonist, and only 1.25-fold less sensitive to authentic NO (all P < .05). In contrast, acute inhibition of CuZnSOD with 10 mM diethyldithiocarbamate produced a more uniform reduction in sensitivity to acetylcholine (8-fold), A23187 (10-fold), and NO (4-fold; all P < .001). Cu-deficient animals demonstrated a 2.5-fold increase in plasma-esterified F2-isoprostanes, a stable marker of lipid peroxidation, that correlated inversely with arterial relaxation to acetylcholine (R = -.83; P < .0009) but not A23187 or authentic NO. From these findings, we conclude that chronic inhibition of CuZnSOD inhibits EDNO-mediated arterial relaxation through two mechanisms, one being direct inactivation of NO and the other being lipid peroxidation that preferentially interrupts receptor-mediated stimulation of EDNO. PMID- 9409285 TI - Conformational changes in apolipoprotein B modulate intracellular assembly and degradation of ApoB-containing lipoprotein particles in HepG2 cells. AB - The linkage between the conformation of apolipoprotein B100 (apoB) and the intracellular assembly and degradation of apoB-containing lipoproteins was investigated in the present study. Disruption of disulfide bond formation in newly synthesized apoB molecules through the use of the reducing agent DTT resulted in a decrease in the secretion of apoB-containing lipoproteins from HepG2 cells compared with control cells. The synthesis of total apoB (apoB100 plus nascent chains), as well as a number of control proteins, such as albumin and alpha 1-antitrypsin, was decreased significantly in DTT-treated cells. However, the intracellular accumulation of full-length apoB100 molecules was not inhibited in the presence of DTT. Subcellular fractionation indicated that apoB molecules isolated from the microsomes of DTT-treated cells had an increased association with the microsomal membrane compared with apoB isolated from untreated cells. Analysis of the distribution of apoB-containing lipoproteins from the lumen of isolated microsomes demonstrated that in the presence of DTT, there was a shift in the distribution, such that there was a decrease in the formation of HDL-sized (lipid-poor) apoB-containing lipoproteins and a decrease in the formation of LDL/VLDL apoB particles. Alterations in apoB conformation and their impact on degradation were also investigated by using DTT and by inhibiting N-linked glycosylation with tunicamycin. DTT appeared to change the rate and pattern of apoB degradation. Degradation was accelerated in both intact and permeabilized HepG2 cells. ApoB degradation occurred in DTT-treated permeabilized cells without the usual generation of the 70-kD and 335-kD fragments and was largely N-acetyl-leucyl-leucyl-norleucinal (ALLN) insensitive. In tunicamycin treated cells, DTT further accelerated the degradation of unglycosylated apoB. Overall, the data suggest that the misfolding of apoB may prevent the proper association of apoB with lipids, resulting in impairment of the assembly of mature apoB-containing lipoproteins. Alteration in the conformation of apoB also appears to alter the degradation pathway of apoB, such that the protein is degraded through a pathway that is at least in part ALLN insensitive. PMID- 9409286 TI - Plasma LDL oxidation leads to its aggregation in the atherosclerotic apolipoprotein E-deficient mice. AB - Two major modifications of low density lipoprotein (LDL) that can lead to macrophage cholesterol accumulation and foam cell formation include its oxidation and aggregation. To find out whether these modifications can already occur in vivo in plasma and whether they are related to each other, the oxidation and aggregation states of plasma LDL were analyzed in the apolipoprotein E-deficient (E degree) transgenic mice during their aging (and the development of atherosclerosis), in comparison to plasma LDL from control mice. Plasma LDL from the E degree mice was already minimally oxidized at 1 month of age in comparison to control mice LDL, and it further oxidized with age in the E degree mice but not in the control mice. At 6 months of age, the contents of the E degree mice LDL-associated cholesteryl ester hydroperoxides, thiobarbituric acid reactive substances, and conjugated dienes were higher by two, three, and twofold, respectively, in comparison to LDL from the young, 1-month-old E degree mice. We also investigated the LDL aggregation state in E degree mice. In the young E degree mice, LDL oxidation was shown in comparison to control mice, but in both groups of young mice their LDL was not aggregated. In the E degree mice, however, the LDL aggregation state substantially increased with age, by as much as 125% at 6 months of age compared to the 1-month-old mice, whereas no significant aggregation could be detected in plasma LDL from control mice at the same age. To question the possible effect of LDL oxidation on its subsequent aggregation, LDL oxidation was induced by either copper ions, or by the free radical generator 2,2 azobis-2-amidinopropane hydrochloride, or by hypochlorite. All these oxidative systems led to LDL oxidation (to different degrees) and resulted in a similar, substantial LDL aggregation. These oxidation systems also enhanced the susceptibility of LDL to aggregation (induced by vortexing) by 23%, 28%, or 40%, respectively. To further analyze the relationships between the lipoprotein oxidation and its aggregation, LDL (0.1 mg of protein/mL) was incubated with 5 mumol/L CuSO4 at 37 degrees C in the absence or presence of the antioxidant, vitamin E (25 mumol/L). In the absence of vitamin E, a time-dependent increment in LDL oxidation was noted, which reached a plateau after 2 hours of incubation. LDL aggregation, however, only started at this time point and reached a plateau after only 5 hours of incubation. In the presence of vitamin E, both LDL oxidation and its aggregation were reduced at all time points studied. We extended the vitamin E study to the in vivo situation, and the effect of vitamin E supplementation to the E degree mice (50 mg.kg-1.d-1 for a 3-month period) on their plasma LDL oxidation and aggregation states was studied. Vitamin E supplementation to these mice resulted in a 35% reduction in the LDL oxidation state and in parallel, the LDL aggregation state was also reduced by 23%. These reductions in LDL oxidation and aggregation states were accompanied by a 33% reduction in the aortic lesion area, in comparison to nontreated E degree mice. We conclude that in E degree mice, LDL oxidation, which already took place in the plasma, can lead to the lipoprotein aggregation. These modified forms of LDL were shown to be taken up by macrophages at an enhanced rate, leading to foam cell formation. Thus, the use of an appropriate antioxidant can inhibit the formation of both atherogenic forms of LDL. PMID- 9409287 TI - Genotype distribution of estrogen receptor polymorphisms in men and postmenopausal women from healthy and coronary populations and its relation to serum lipid levels. AB - The cardiovascular protective effects of estrogen are known to be mediated by its beneficial effects on lipid metabolism and its direct actions on the vessel wall. The latter can be mediated by a specific receptor for estrogen present on smooth muscle cells and endothelial cells. The gene for the receptor (the classic estrogen receptor [ER]) has three known polymorphisms, Pvu II, Xba I, and B variant polymorphisms, which are reportedly associated with receptor expression and altered receptor function and with some disorders including breast cancer, hypertension, and spontaneous abortion. However, the significance of genetic variations of the ER in vascular diseases has not been reported. We have examined the association between coronary artery disease (CAD) and the three polymorphisms in ER. Genotypes (P1/P2, X1/X2, and B-wild type/B-variant type) were determined in 87 men and postmenopausal women with myocardial infarction or angina pectoris whose lesions were confirmed by coronary angiography, as well as from 94 control individuals from the general population with no coronary heart disease and normal resting ECG. For B-variant polymorphism, all individuals examined had B-wild type, which contrasts with the reported allele frequency for B-variant type (0.1) in the white population. Genotype distributions and allele frequencies of Pvu II or Xba I polymorphisms were not significantly different between control subjects and patients (P > .05 for Pvu II or Xba I genotypes; P > .05 for Pvu II or Xba I allele frequencies). When the allele frequencies were analyzed separately by sex, there was still no statistically significant difference for both polymorphisms (P > .05 for men; P > .05 for women). No association was found between the polymorphisms and the angiographic severity of CAD. Total cholesterol, triglyceride, or HDL-cholesterol levels were not significantly different among ER genotypes. These findings suggest that the three polymorphisms in ER are not associated with the prevalence and severity of CAD and that the polymorphisms are unrelated to the serum lipid levels in control subjects and patients. PMID- 9409288 TI - Two intracellular signaling pathways for activation of protein kinase C are involved in oxidized low-density lipoprotein-induced macrophage growth. AB - Recent studies demonstrated that oxidized LDL (Ox-LDL) induces macrophage growth in vitro. The present study was undertaken to elucidate the intracellular signaling pathways for macrophage growth. Ox-LDL initiated a rapid and transient rise in intracellular free calcium ion and induced activation of membrane protein kinase C (PKC). Pertussis toxin completely inhibited the Ox-LDL-induced rise in free calcium ion and significantly inhibited macrophage growth by 50%. Moreover, PKC inhibitors calphostin C and H-7 significantly inhibited Ox-LDL-induced macrophage growth by 80%. On the other hand, phospholipase A2-treated acetylated LDL did not induce a rise in calcium but significantly activated PKC and led to significant macrophage growth that was significantly inhibited by calphostin C by 90%. These results suggest the presence of two intracellular signaling pathways for activation of PKC, a rise in calcium that was mediated by pertussis toxin sensitive G protein and the internalization of lysophosphatidylcholine through the scavenger receptors. These two pathways may play an important role in Ox-LDL induced macrophage growth. PMID- 9409289 TI - Apolipoprotein A-IFIN (Leu159-->Arg) mutation affects lecithin cholesterol acyltransferase activation and subclass distribution of HDL but not cholesterol efflux from fibroblasts. AB - We showed earlier that the apolipoprotein A-I Leu159-->Arg mutation (apoA-IFin) results in dominantly inherited hypoalphalipoproteinemia. In the present study we investigated the effect of the apoA-IFin mutation on lipoprotein profile, apoA-I kinetics, lecithin:cholesterol acyltransferase (LCAT) activation, and cholesterol efflux in vitro. Carriers (n = 9) of the apoA-IFin mutation exhibited several lipoprotein abnormalities. The serum HDL cholesterol level was diminished to 20% of normal, and nondenaturing gradient gel electrophoresis of HDL showed disappearance of particles at the 9.0- to 12-nm size range (HDL2-type) and the presence of small 7.8- to 8.9-nm (mostly HDL3-type) particles only. HDL3-type particles from both the mutation carriers and nonaffected family members were similarly converted to large, HDL2-type particles by phospholipid transfer protein in vitro. Studies on apoA-I kinetics in four affected subjects favored accelerated catabolism of apoA-I. Experiments with reconstituted proteoliposomes showed that the capacity of apoA-IFin protein to activate LCAT was reduced to 40% of that of the wild-type apoA-I. The impact of the apoA-IFin protein on cholesterol efflux was examined in vitro using [3H]cholesterol-loaded human fibroblasts and three different cholesterol acceptors: (1) total HDL, (2) total apoA-I combined with phospholipid, and (3) apoA-I isoform (apoA-IFin or wild-type apoA-I isoform 1) combined with phospholipid. ApoA-IFin did not impair phospholipid binding or cholesterol efflux from fibroblasts to any of the acceptors used. Only one of the nine apoA-IFin carriers appears to have evidence of clinically manifested atherosclerosis. In conclusion, although the apoA-IFin mutation does not alter the properties of apoA-I involved in promotion of cholesterol efflux, its ability to activate LCAT in vitro is defective. In vivo, apoA-IFin was found to be associated with several lipoprotein composition rearrangements and increased catabolism of apoA-I. PMID- 9409290 TI - Cholesterol-mediated changes of neutral cholesterol esterase activity in macrophages. Mechanism for mobilization of cholesteryl esters in lipid droplets by HDL. AB - Cholesteryl esters (CE) in lipid droplets undergo a continual cycle of hydrolysis and reesterification by neutral cholesterol esterase (N-CEase) and acyl CoA:cholesterol acyltransferase (ACAT), respectively. The mechanism by which HDL mobilizes CE from lipid droplets in J774 A.1 cells was investigated, focusing on N-CEase activity. We asked whether HDL enhances the activity and, if so, what signals induce the change of the activity. An incubation of cells with HDL enhanced the decline of cholesteryl-[l-14C]-oleate in foam cells and increased N CEase activity in the supernatant of cell homogenate in a concentration-dependent manner, whereas incubation with LDL decreased the activity. In addition, N-CEase activity was fivefold higher when cells were cultured in 10% lipoprotein deficient serum (LPDS) medium (2 micrograms cholesterol/mL) than when cultured in 10% fetal calf serum medium (31 micrograms cholesterol/mL), suggesting that changes in N-CEase activity are mediated by cholesterol. An addition of cholesterol (0 to 30 micrograms/mL) in LPDS medium markedly inhibited N-CEase activity with a concomitant increase in cellular cholesterol concentration. This inhibitory effect of cholesterol was also observed in mouse peritoneal macrophages. In vitro addition of cholesterol did not affect N-CEase activity. Treatment of cells with HMG-CoA reductase inhibitors enhanced N-CEase activity, whereas ACAT inhibitor decreased the activity. Northern blot analysis of N-CEase mRNA showed that the expression was not altered by the presence of cholesterol in LPDS medium. These results suggest that cholesterol downregulates N-CEase activity, probably through cholesterol-dependent appearance of some factors. PMID- 9409291 TI - Suppression of lipid transfer inhibitor protein activity by oleate. A novel mechanism of cholesteryl ester transfer protein regulation by plasma free fatty acids. AB - Cholesteryl ester transfer protein (CETP) mediates the interlipoprotein exchange of cholesteryl ester (CE) and triglyceride. A second plasma protein, lipid transfer inhibitor protein (LTIP), binds to lipoproteins and inhibits CETP activity by displacing CETP from the lipoprotein surface. Since free fatty acids (FFAs) enhance the binding of CETP to lipoproteins, we have examined the possible role of FFAs in modulating LTIP activity. Partially purified CETP, LTIP, and lipoproteins were incubated with 0 to 30 mumol/L sodium oleate, and the transfer of CE between a labeled donor lipoprotein and a given acceptor lipoprotein was measured. Without LTIP, oleate stimulated CETP-mediated CE transfer between VLDL, LDL, and HDL up to threefold. This stimulation was unique in both magnitude and oleate concentration dependence for each donor-acceptor lipoprotein pair. In contrast to CETP activity, in transfer reactions involving LDL or VLDL as donor, LTIP activity was suppressed (> 80%) by 10 to 15 mumol/L oleate. LTIP activity in transfer reactions with HDL as donor was less sensitive. Similar results to these were observed when lipid transfer reactions were measured in the total lipoprotein fraction isolated from FFA-enriched plasma. The FFA content of lipoproteins was strongly influenced by the concentration of FFA in plasma; lipoprotein FFA levels sufficient to suppress LTIP activity by 50% to 100% were achieved in plasma containing 0.8 to 1.0 mmol/L FFA. We conclude that LTIP may be functionally inactive during periods of transient elevations of plasma FFA levels, such as during postprandial lipemia or overnight fasting, or chronically suppressed in disease states in which plasma FFA levels are increased. The suppression of LTIP activity by FFA allows for maximum CETP-mediated lipid transfer between all lipoproteins, including lipid transfer reactions involving LDL that are normally preferentially suppressed by LTIP. PMID- 9409292 TI - Antithrombotic efficacy of inactivated active site recombinant factor VIIa is shear dependent in human blood. AB - Several studies have indicated a profound role for factor VII(a) [FVII(a)] in venous and arterial thrombogenesis. In the present study, we quantified the inhibitory efficacy of dansyl-glutamyl-glycyl-arginyl-recombinant FVIIa (DEGR rFVIIa) on acute thrombus formation. Thrombus formation was elicited by immobilized tissue factor (TF) in a parallel-plate perfusion chamber device at blood flow conditions characterized by wall shear rates of 100 S-1 (veins) and 650 S-1 (medium-sized healthy arteries). Native human blood was drawn directly from an antecubital vein by a pump into a heparin-coated mixing device in which DEGR-rFVIIa (0.09 to 880 nmol/L final plasma concentration) or buffer was mixed homogeneously with flowing blood. Subsequently, the blood was passed over a plastic coverslip coated with TF and phospholipids in the parallel-plate perfusion chamber. Fibrin deposition, platelet-fibrin adhesion, and platelet thrombus volume triggered by this surface were measured by morphometry. DEGR rFVIIa inhibited thrombus formation in a dose-dependent manner, but the efficacy was shear rate dependent. At a wall shear rate of 100 S-1, the IC50 (50% inhibition) was 30 nmol/L, whereas at 650 S-1, the IC50 was 0.6 nmol/L. Binding studies to immobilized TF under flow conditions using surface plasmon resonance revealed a significantly higher on-rate for DEGR-rFVIIa and FVIIa than for FVII, 2.8 x 10(5), 2.6 x 10(5), and 1.8 x 10(5) M-1 S-1, respectively. This indicates that a contributing factor to the shear-dependent efficacy may be a differential importance of on-rates at arterial and venous blood flow conditions. PMID- 9409293 TI - The impact of atherosclerotic arterial remodeling on percentage of luminal stenosis varies widely within the arterial system. A postmortem study. AB - Luminal stenosis can be based on large atherosclerotic plaques in compensatory enlarged segments or on relatively little plaques in shrunken segments. In the present study, the contribution of plaque formation and remodeling to luminal narrowing was compared among six types of arteries prone to symptomatic atherosclerosis. Cross-sections (n = 5195) were obtained at regular intervals from 329 arteries. For each artery, the cross-section that contained the least amount of plaque was considered to be the reference. For each cross-section, the percentage of lumen area decrease was expressed as a percentage of the lumen area at the reference site (luminal stenosis). Similarly, the area encompassed by the internal elastic lamina (IEL area) was expressed as a percentage of the IEL area at the reference site (relative IEL area). All cross-sections were categorized in three groups: relative IEL area > 105% (enlargement), 95% to 105% (no remodeling), and < 95% (shrinkage). The prevalence of enlargement (50% to 75%) was significantly higher compared with shrinkage (8% to 25%). Shrinkage was observed most frequently in the femoral arteries (25%) and infrequently in the renal arteries (8%). For all types of arteries, the relative IEL area correlated negatively with luminal stenosis (P < .001). Regression analysis of relative IEL area on luminal stenosis, however, showed significant differences in the first order regression coefficients among artery types. On average, plaque increase was more compensated for by enlargement in the coronary, common carotid, and renal arteries compared with the arteries obtained from the lower extremities. Anatomic regional differences were observed in the impact of arterial wall remodeling on percent luminal stenosis in de novo atherosclerotic lesions. PMID- 9409295 TI - Effects of oral and transdermal estrogen/progesterone regimens on blood coagulation and fibrinolysis in postmenopausal women. A randomized controlled trial. AB - Postmenopausal hormone replacement therapy is associated with a reduction in the incidence of coronary heart disease. However, inconclusive results have been reported with respect to the risk of stroke, and recent studies consistently showed an increased risk of venous thromboembolism in postmenopausal women using oral estrogen. There are surprisingly few interventional studies to assess the true effects of estrogen-progestin regimens on blood coagulation and fibrinolysis, and the impact of the route of estrogen administration on hemostasis has not been well documented. Therefore, we investigated the effects of oral and transdermal estradiol/progesterone replacement therapy on hemostatic variables. Forty-five healthy postmenopausal women, aged 45 to 64 years, were assigned randomly to one of the three following groups: cyclic oral or transdermal estradiol, both combined with progesterone, or no hormonal treatment. Hemostatic variables were assayed at baseline and after a 6-month period. Pairwise differences in the mean change between the three groups were compared using nonparametric tests. Oral but not transdermal estradiol regimen significantly increased the mean value of prothrombin activation peptide (F1 + 2) and decreased mean antithrombin activity compared with no treatment. Differences in fragment F1 + 2 levels between active treatments were significant. The oral estrogen group was associated with a significant decrease in both mean tissue type plasminogen (t-PA) concentration and plasminogen activator inhibitor (PAI-1) activity and a significant rise in global fibrinolytic capacity (GFC) compared with the two other groups. A transdermal estrogen regimen had no significant effect on PAI-1, t-PA, and GFC levels. There were no significant changes in mean values of fibrinogen, factor VII, von Willebrand factor, protein C, fibrin D dimer, and plasminogen between and within the three groups. We conclude that oral estrogen/progesterone replacement therapy may result in coagulation activation and increased fibrinolytic potential, whereas opposed transdermal estrogen appears without any substantial effects on hemostasis. Whereas these results may account for an increased risk of venous thromboembolism in users of oral postmenopausal estrogen, they emphasize the potential importance of the route of estrogen administration in prescribing hormone replacement therapy to postmenopausal women, especially to those at high risk of thrombotic disease. PMID- 9409294 TI - Cafestol, the cholesterol-raising factor in boiled coffee, suppresses bile acid synthesis by downregulation of cholesterol 7 alpha-hydroxylase and sterol 27 hydroxylase in rat hepatocytes. AB - Consumption of boiled coffee raises serum cholesterol levels in humans. The diterpenes cafestol and kahweol in boiled coffee have been found to be responsible for the increase. To investigate the biochemical background of this effect, we studied the effects of cafestol and a mixture of cafestol/kahweol/isokahweol (48:47:5 w/w) on bile acid synthesis and cholesterol 7 alpha-hydroxylase and sterol 27-hydroxylase in cultured rat hepatocytes. Dose dependent decreases of bile acid mass production and cholesterol 7 alpha hydroxylase and sterol 27-hydroxylase activity were found, showing a maximal reduction of -91%, -79%, and -49% respectively, at a concentration of 20 micrograms/mL cafestol. The decrease in 7 alpha-hydroxylase and 27-hydroxylase activity paralleled well the suppression of the respective mRNAs, being -79% and 77%, and -49% and -46%, respectively, at 20 micrograms/mL cafestol. Run-on data showed a reduction in 7 alpha-hydroxylase and 27-hydroxylase gene transcriptional activity after incubation with cafestol. The mixture of cafestol/kahweol/isokahweol was less potent in suppression of bile acid synthesis and cholesterol 7 alpha-hydroxylase. Cafestol (20 micrograms/mL) had no effect on lithocholic acid 6 beta-hydroxylase mRNA, another enzyme involved in bile acid synthesis. LDL-receptor, HMG-CoA reductase, and HMG-CoA synthase mRNAs were significantly decreased by cafestol (-18%, -20%, and -43%, respectively). We conclude that cafestol suppresses bile acid synthesis by downregulation of cholesterol 7 alpha-hydroxylase and of, to a lesser extent, sterol 27-hydroxylase in cultured rat hepatocytes, whereas kahweol and isokahweol are less active. We suggest that suppression of bile acid synthesis may provide an explanation for the cholesterol-raising effect of cafestol in humans. PMID- 9409297 TI - Loss of arterial dilation in the reendothelialized area of the flow-loaded rat common carotid artery. AB - We have investigated regenerated endothelial cells and their possible contribution to arterial dilation in response to increased blood flow in rat common carotid artery (CCA). After endothelial denudation using a balloon catheter in the left CCA, an arteriovenous shunt was constructed between the left CCA and the left external jugular vein at 20 mm distal from the orifice in the denuded group. Animals that were given the arteriovenous shunt without denudation were used to form the nondenuded group. The blood flow rate in the left CCA was increased by sixfold after operation in the denuded group. We observed that endothelial cells were gradually regenerated from the orifice to the distal area and that the reendothelialized area after 4 to 8 weeks was approximately one third of the left CCA (5.31 +/- 1.49 mm at 4 weeks, 5.47 +/- 1.56 mm at 8 weeks). In the reendothelialized area of the left CCA after 4 to 8 weeks, the lumen diameter was significantly smaller than that of the nondenuded group and showed no significant difference from age-matched nonsurgical animals. The intimal and medial thickening, which would result in arterial stenosis in the reendothelialized area, was not observed in the denuded group, although the denuded control showed significant intimal thickening. From these results, we conclude that regenerated endothelial cells reduce intimal thickening but do not respond to increased blood flow to dilate the artery. PMID- 9409296 TI - Induction and role of NO synthase in hypotensive hepatic failure. AB - Nitric oxide (NO) plays an important role in the physiological and pathophysiological control of the vascular system. NO is synthesized by isoforms of the enzyme NO synthase (NOS). Hepatic failure is complicated by hypotension, low systemic vascular resistance, and resistance to vasoconstrictor drugs. The potential role of NO in these abnormalities was investigated by using in vitro pharmacological interventions on hepatic arteries obtained from both donor and recipient patients at the time of liver transplantation. The presence of NOS mRNA was investigated by reverse transcription polymerase chain reaction (RT-PCR) with primers designed from human endothelial NOS (eNOS) and inducible NOS (iNOS) cDNA sequences. Arteries from patients with hepatic failure had an impaired constrictor response to phenylephrine compared with those of donor arteries. The constrictor effect of phenylephrine was potentiated by NG-monomethyl-L-arginine, an inhibitor of NOS, which had no effect in donor control arteries. RT-PCR identified human eNOS mRNA in donor and recipient arteries and human iNOS mRNA in recipient arteries only. Induction of NOS in the vasculature with subsequent NO induced vasodilatation may therefore contribute to the hemodynamic abnormalities observed in hepatic failure and potentially in other pathologies associated with endotoxemia. Whether selective inhibitors of iNOS will improve hemodynamic control or clinical outcome in these conditions requires further study. PMID- 9409299 TI - Shear stress gradients remodel endothelial monolayers in vitro via a cell proliferation-migration-loss cycle. AB - Wall shear stress has been implicated in the genesis of atherosclerosis because a strong correlation exists between the location of developing arterial lesions and regions where particular gradients in stress occur. Studying the behavior of endothelial cells in such regions may contribute to our understanding of the disease etiology. We report the detailed migratory history of endothelial cells subjected to large shear stress gradients caused by a surface protuberance in an in vitro model system. The history of cell migration, cell division, and cell loss from the surface was continuously monitored in confluent human umbilical vein endothelial cell monolayers for 48 hours after the onset of flow. Individual cells were tracked using time-lapse video microscopy. In contrast to a uniform laminar flow field in which cells were observed to continually rearrange their relative position with no net migration, in a disturbed flow field there was a net migration directed away from the region of high shear gradient. This organized migration pattern under disturbed flow conditions was accompanied by more than a twofold increase in cell motility. In addition, cell division increased in the vicinity of the flow separation (maximum shear stress gradient of 34 dyne/cm2 per mm) whereas cell loss was increased upstream and downstream in the regions where the shear gradient diminishes. These data suggest a steady cell proliferation-migration-loss cycle and indicate that local shear stress gradient may play a key role in the morphological remodeling of the vascular endothelium in vivo. PMID- 9409298 TI - Comparison of the genetic defect with LDL-receptor activity in cultured cells from patients with a clinical diagnosis of heterozygous familial hypercholesterolemia. The Familial Hypercholesterolaemia Regression Study Group. AB - In this study we have analyzed the genetic defect in 42 patients with a diagnosis of heterozygous familial hypercholesterolemia (FH) by Southern blotting, SSCP, and sequencing of PCR-amplified fragments of genomic DNA or sequencing of RT-PCR products from mRNA in cultured cells. The apoB Arg3500Gln mutation was identified in five patients. A molecular defect in the LDL-receptor gene was confirmed in 23 patients; 16 of these mutations have not been described before. No defect in the coding region, intron:exon junctions or proximal promoter of the LDL-receptor gene or in the region of the apoB gene coding for the LDL-receptor binding domain was found in the remaining 14 patients. LDL-receptor activity and protein content of cultured lymphoblasts from the patients was significantly lower in cells from patients with severe rather than mild LDL-receptor mutations. Cells from four patients with no detectable defect showed reduced LDL receptor activity compared with eight normal cell lines, whereas six others had reduced LDL-receptor activity but LDL-receptor protein content within the normal range. Cells from four patients appeared to have normal LDL-receptor function. Cells from two patients with a defined defect also had LDL-receptor activity within the normal range. The findings demonstrate the problems involved in the genetic diagnosis of FH in patients. PMID- 9409300 TI - Human monocyte-derived macrophages express an approximately 120-kD Ox-LDL binding protein with strong identity to CD68. AB - A protein that specifically binds oxidized LDL (Ox-LDL) has recently been characterized in mouse peritoneal macrophages and identified as macrosialin, a protein with a molecular weight of 95 kD. First, the present work shows that human monocyte-derived macrophages express a membrane protein with a molecular weight of approximately 120 kD that selectively binds Ox-LDL. Second, we tested whether this approximately 120-kD Ox-LDL binding protein had any relation to CD68, the human homologue of macrosialin. The following evidence was obtained to support the role of CD68 as an Ox-LDL binding protein: (1) Ligand blots with Ox LDL and Western blots with Ki-M6, an anti-human CD68 monoclonal antibody, revealed a single band with a molecular weight of approximately 120 kD under reducing and nonreducing condition. (2) The expression patterns of the approximately 120-kD Ox-LDL binding membrane protein and of CD68 paralleled each other during monocyte/macrophage differentiation. (3) Digestion with N glycosidase F demonstrated that both CD68 and the Ox-LDL binding protein are glycoproteins; both showed a similar shift of approximately 18 kD in apparent molecular weight. (4) CD68, probed with monoclonal antibody Ki-M6, and the approximately 120-kD Ox-LDL binding protein were coprecipitated with EMB11, another anti-CD68 antibody. About 5000 molecules of CD68 are expressed on the cell surface of human macrophages. Ligation of 125I-Ki-M6 to cells leads to its internalization and degradation. This capacity would be sufficient to allow for the specific uptake and degradation of Ox-LDL. Taken together, these data support a role for CD68 as a specific Ox-LDL binding protein in human monocyte-derived macrophages. PMID- 9409301 TI - Bone marrow transplantation in apolipoprotein E-deficient mice. Effect of ApoE gene dosage on serum lipid concentrations, (beta)VLDL catabolism, and atherosclerosis. AB - Apolipoprotein E (apoE), a high-affinity ligand for lipoprotein receptors, is synthesized by the liver and extrahepatic tissues, including cells of the monocyte/macrophage lineage. Inactivation of the apoE gene in mice leads to a prominent increase in serum cholesterol and triglyceride levels and the development of premature atherosclerosis. In this study, the role of monocyte/macrophage-derived apoE in lipoprotein remnant metabolism and atherogenesis was assessed. The influence of apoE gene dosage on serum lipid concentrations was determined by transplantation of homozygous apoE-deficient (apoE-/-), heterozygous apoE-deficient (apoE+/-), and wild-type (apoE+/+) bone marrow in homozygous apoE-deficient mice. The concentration of apoE detected in serum was found to be gene dosage dependent, being 3.52 +/- 0.30%, 1.87 +/- 0.17%, and 0% of normal in transplanted mice receiving either apoE+/+, apoE+/-, or apoE-/- bone marrow, respectively. These low concentrations of apoE nevertheless dramatically reduced serum cholesterol levels owing to a reduction of VLDL and, to a lesser extent, LDL, while HDL levels were slightly raised. After 4 months on a "Western-type" diet, atherosclerosis was evidently reduced in mice transplanted with apoE+/+ bone marrow, compared with control transplanted mice. To study the mechanism of the lipoprotein changes on bone marrow transplantation, the in vivo turnover of autologous serum (beta)VLDL was studied. The serum half-life of (beta)VLDL in transplanted mice, compared with control apoE-deficient mice, was shortened mainly as a consequence of an increased recognition and uptake by the liver. Analysis of the relative contribution of the liver parenchymal cells, endothelial cells, and Kupffer cells (liver tissue macrophages) indicated an increased uptake by parenchymal cells, while the relative contribution to Kupffer cells was decreased. In conclusion, macrophage derived apoE can dose-dependently reduce hypercholesterolemia in apoE-deficient mice owing to increased recognition and uptake of (beta)VLDL by parenchymal liver cells, leading to a decreased susceptibility to atherosclerosis. PMID- 9409302 TI - Familial hypercholesterolemia in the Finnish north Karelia. A molecular, clinical, and genealogical study. AB - A specific mutation termed FH-North Karelia [FH-NK] accounts for almost 90% of familial hypercholesterolemia [FH] cases in the Finnish North Karelia, with a population of about 180,000. Extensive search for its presence in the entire North Karelia province revealed 340 carriers of this mutation. Other mutations of the LDL receptor [LDLR] gene accounted for 67 cases of heterozygous FH. This gives a minimum FH prevalence of 1 in 441 inhabitants in North Karelia, with the highest density of patients in the Polvijarvi commune (1 in 143 inhabitants). Old parish records, confirmation records, and tax records were used to track a common ancestor for most of the present-day North Karelian FH-NK patients in the village of Puso, located within an area where the FH prevalence today is the highest. DNA analysis indicated that 2% of the subjects aged 1 to 25 years would have been diagnosed as false-negative and 7% as false-positive FH patients on the basis of LDL cholesterol [LDL-C] determinations alone. Common genetic variations of apolipoprotein E [apoE], XbaI, polymorphism of apolipoprotein B [apoB], and PvuII polymorphism of the intact LDLR allele contributed little to serum lipid variation in established carriers of the FH-NK allele, although apoE2/4 genotype and the presence of the PvuII restriction site tended to be associated with relatively low LDL-C levels. Coronary heart disease (CHD) was present in 65 (30%) out of the 179 FH gene carriers aged > or = 25 years, and 19 individuals had a previous history of acute myocardial infarction (AMI). The average age (mean +/- SD) at onset of CHD was 42 +/- 7 years for males and 48 +/- 11 years for females (P < .05). In stepwise logistic regression analysis carried out in carriers of the FH-NK allele, age, gender, smoking, and apoE allele E2 all emerged as independent determinants of risk of CHD or AMI. It may be concluded that the relatively high prevalence of FH patients in North Karelia province provides a unique founder population in which genetic and nongenetic factors modifying the course of FH can be effectively investigated. PMID- 9409303 TI - Thrombomodulin expression in bovine brain capillaries. Anticoagulant function of the blood-brain barrier, regional differences, and regulatory mechanisms. AB - Thrombomodulin (TM), a key cofactor of the TM-protein C pathway, is of major biologic significance for the antithrombotic properties of endothelial cells. Yet, there is uncertainty whether TM is expressed in brain and what mechanisms govern brain endothelial anticoagulant activity. In this study, bovine brain capillaries were used as an in vitro model of the blood-brain barrier to determine factors involved in the regulation of TM expression in cerebral vasculature. Quantitative competitive-polymerase chain reaction assay revealed significant regional differences in the amount of brain capillary TM mRNA, i.e., cortical > cerebellar > pontine, consistent with the reverse transcription polymerase chain reaction findings in which the abundance of TM mRNA was analyzed relative to beta-actin mRNA. Regional differences in TM mRNA brain capillary level correlated well with differences in protein C activation. The TM mRNA and activity were not detectable in brain parenchyma. Pathogenic mediators of ischemic stroke, interleukin 1 beta (10 U/mL), and tumor necrosis factor alpha (10 U/mL), produced a time-dependent decrease in brain capillary TM mRNA (t1/2 of 2.1 and 3.9 hours, respectively) and reduced endothelial TM activity. Incubation of brain capillaries with retinoic acid (10 mumol/L) and dibutyryl cAMP (3 mmol/L) resulted in a 4-fold increase in TM mRNA at 4 and 8 hours, respectively, followed by an increase in protein C activation. We conclude that TM at the blood brain barrier is likely to be an important physiologic anticoagulant in brain microcirculation. Its downregulation by cytokines may contribute to ischemic brain damage and potentially could be counteracted by retinoic acid and cAMP. PMID- 9409304 TI - Genetic contribution of the endothelial constitutive nitric oxide synthase gene to plasma nitric oxide levels. AB - Nitric oxide (NO) has an important physiological role in regulating vascular tone and is also relevant to many pathological processes including hypertension and atherosclerosis. Endothelial constitutive nitric oxide synthase (ecNOS) is the key enzyme in determining basal vascular wall NO production. We used a combination of maximum-likelihood-based statistical genetic methods to explore the contributions of the ecNOS gene and other unmeasured genes to basal NO production measured by its metabolites (NOx: nitrite and nitrate) in 428 members of 108 nuclear families. Our initial quantitative genetic analysis estimated that approximately 30% of the variance in fasting NOx levels is due to genes (chi 2(1) = 16.04, P = .000062). Complex segregation analysis detected the effects of both a single locus and residual polygenes on NOx levels, and measured genotype analysis showed that plasma NOx levels in those homozygous for the rare allele (64.9 +/- 7.8 mumol/L) were significantly higher (P = .000242) than those homozygous for the common allele (30.2 +/- 3.1 mumol/L). The results of the variance component linkage analysis were consistent with linkage of a quantitative trait locus in or near the ecNOS gene to variation in plasma NOx levels (P = .0066). While many environmental factors have been shown to alter transiently plasma NOx levels, our study is the first to identify a substantial effect of the ecNOS locus on the variance of plasma NOx, i.e. basal NO production. This finding may be relevant to atherogenesis and NO-related disorders. PMID- 9409305 TI - Binding properties and inhibition of platelet aggregation by a monoclonal antibody to CD31 (PECAM-1). AB - CD31/platelet endothelial cell adhesion molecule 1 (PECAM-1) is expressed on platelets, endothelial cells, myeloid cells, monocytes, and certain lymphocyte subsets. It has been shown that CD31 is involved in the homophilic and heterophilic cellular adhesion and leukocyte transendothelial migration, but little is known about the role of CD31 in platelet functions. Previously we have shown that monoclonal antibody (MAb) AAP2 produced in our laboratory bound to a 110-kD platelet antigen and gave an enhanced binding to activated platelet membrane. In this study we demonstrated that platelet lysate depleted of the antigen through adsorption by an AAP2-solidified affinity column was bound by MAbs against CD62 and CD42 but not by MAb 5.6E against CD31 or AAP2 on the immunoblot. Rabbit antibodies against CD31 completely inhibited the binding of AAP2 to platelets in the flow cytometry analysis. This indicates that AAP2 is specifically against CD31. 125I-labeled AAP2 bound to resting platelets with 5587 +/- 1765 sites/platelet and a Kd of 1 nmol/L and to thrombin-activated platelets with 17,625 +/- 4865 sites/platelet and a Kd of 0.24 nmol/L. Addition of 10 micrograms/mL AAP2 inhibited the aggregation induced by 4 mumol/L ADP by 78.6%, 6 mumol/L epinephrine by 79.4%, 1 microgram/mL collagen by 78.7%, and 0.25 U/mL thrombin by 29%. The platelet aggregation was completely restored when higher concentration of agonists were used. MAb 5.6E did not have any effect on platelet aggregation. These results suggest that AAP2 binds to a special epitope of CD31 on platelets and that CD31 is involved in platelet aggregation. PMID- 9409306 TI - Antibodies against cardiolipin and oxidatively modified LDL in 50-year-old men predict myocardial infarction. AB - Autoantibodies against oxidatively modified low-density lipoproteins (oxLDL) and cardiolipin occur in patients with vascular diseases, including atherosclerosis. The ability of such antibodies to predict myocardial infarction (MI) was investigated in a prospective nested case-control study in which healthy 50-year old men were followed up for 20 years. Raised levels of antibodies against oxLDL and cardiolipin at 50 years of age correlated positively with the incidence of MI and mortality related to MI 10 to 20 years later. IgG and IgA antibodies against cardiolipin were associated with MI between 50 to 60 years of age and IgG and IgA antibodies against oxLDL with MI at 60 to 70 years of age. Moreover, higher antibody levels were noted in those who died from acute MI in comparison to those who survived. The predictive power of IgA and IgG antibodies was strong and largely independent of that of other strong risk factors. In conclusion, raised levels of antibodies against oxLDL and cardiolipin may predict MI and MI-related death. PMID- 9409307 TI - Dopamine as a novel antimigration and antiproliferative factor of vascular smooth muscle cells through dopamine D1-like receptors. AB - Vascular smooth muscle cell (VSMC) migration and proliferation are believed to play key roles in atherosclerosis. To elucidate the role of vascular dopamine D1 like receptors in atherosclerosis, the effects of dopamine and specific D1-like agonists SKF 38,393 and YM 435 on platelet-derived growth factor (PDGF) BB mediated VSMC migration and proliferation were studied. We observed that cells stimulated by PDGF-BB (5 ng/mL), showed increased migration and proliferation. These effects were prevented by coincubation with dopamine, SKF 38,393, or YM 435 (1 to 10 mumol/L), and this prevention was reversed by Sch 23,390 (1 to 10 mumol/l), a specific D1-like antagonist. These actions are mimicked by forskolin (1 to 10 mumol/L), a direct activator of adenylate cyclase and 8-bromo-cAMP at 0.1 to 1 mmol/L and are blocked by a specific protein kinase A inhibitor, N-[2-(p bromocinnamylamino)ethyl]-5-isoquinoline-sulfonamide (H 89), but not blocked by its negative control, N-[2-(N-formyl)-p-chlorociannamylamino)ethyl]-5 isoquinoline sulfonamide (H 85). PDGF-BB (5 ng/mL)-mediated activation of phospholipase D, protein kinase C, and mitogen activated protein kinase activity were significantly suppressed by coincubation with dopamine. These results suggest that vascular D1-like receptor agonists inhibit migration and proliferation of VSMC, possibly through protein kinase A activation and suppression of activated phospholipase D, protein kinase C, and mitogen-activated protein kinase activity. PMID- 9409308 TI - Upregulation of connexin43 gap junctions between smooth muscle cells after balloon catheter injury in the rat carotid artery. AB - Phenotypic transformation of smooth muscle cells (SMCs) to the synthetic state in vitro and in human coronary atherosclerosis is reported to be associated with upregulation of connexin43 gap junctions. To determine whether cellular interactions mediated by gap junctions participate in the phenotypic transformation of SMCs in arterial injury and disease in general and to establish the spatial and temporal pattern of any such change in relation to neointimal development, we investigated SMC connexin43 gap junction expression during vascular healing in the rat carotid artery after balloon catheter injury. Quantitative immunoconfocal microscopy was applied to localize and to quantify connexin43 gap junctions 1, 3, 9, and 14 days after injury. Parallel studies were conducted by electron microscopy (direct morphological demonstration of SMC gap junctions) and immunoconfocal microscopy (localization of altered actin expression). Synthetic-state SMCs in the neointima (first apparent from 9 days postinjury) revealed abundant expression of gap junctions, with levels of immunodetectable connexin43 threefold greater than those of medial cells. However, the first detectable changes were found in the media, before neointimal formation; at 1 to 3 days postinjury, an increase in SMC gap junction expression was apparent in the innermost (subluminal) zone, the major site from which the cells subsequently found in the neointima are recruited. We conclude that upregulation of connexin43 gap junctions is intimately linked to SMC phenotypic transition and that interactions mediated by gap junctions may be a hitherto unrecognized contributor to the cellular mechanisms underlying the vascular response to injury. PMID- 9409309 TI - cAMP involvement in the expression of MMP-2 and MT-MMP1 metalloproteinases in human endothelial cells. AB - Matrix metalloproteinases (MMPs) are a multigene family of enzymes secreted by a variety of cells, including human umbilical vein endothelial cells (HUVECs). Because metalloproteinases are potentially destructive agents, their production is tightly controlled at several levels. Rather little is known about the presence and regulation of MMPs in endothelial cells. In this study, we investigated the expression and regulation of MMP-2 and membrane type-matrix metalloproteinase (MT-MMP1), a membrane metalloproteinase strictly related to MMP 2 activation. Zymographic analysis of conditioned medium (CM) of HUVECs showed the presence of gelatinolytic activity mainly at 72 and 64 and 62 kD. The 64- and 62-kD bands, respectively, represent the intermediate and the completely active forms of MMP-2. When HUVECs were treated with forskolin (FK) (100 and 25 mumol/l), there was a decrease in the appearance of the 64 to 62 kDa doublet, suggesting an inhibition of the fully activated form of MMP-2. FK raises intracellular cAMP in HUVECs. The same data were obtained using dibutyryl-cAMP. Northern analysis revealed that the expression of MMP-2 increased slightly after treatment with FK, in contrast with gelatin zymography results. Taking into consideration the mechanism of activation of MMP-2, we tested the hypothesis that this compound could modulate MT-MMP1. As expected, FK was able to decrease MT MMP1 expression. These data correlate with experiments using membranes of FK treated HUVECs and incubated with control CM. Zymography revealed that when CM was incubated with membranes prepared from FK-treated HUVECs, there was a decrease in the appearance of the 64-kDa band, suggesting that the expression of MT-MMP1 was negatively modified. These results correlate with the MT-MMP1 protein level, negatively modified after FK treatment. PMID- 9409310 TI - Plasma levels of nitrite/nitrate and platelet cGMP levels are decreased in patients with atrial fibrillation. AB - Patients with atrial fibrillation have been reported to exhibit abnormal hemostasis. Since nitric oxide (NO) exerts antithrombotic effects and attenuates platelet function, we evaluated two indicators of plasma NO levels, the plasma levels of nitrite and nitrate (NOx), and the levels of cGMP in platelets. We also examined whether indicators of plasma NO levels were associated with abnormalities in parameters related to platelet function, blood coagulation, and fibrinolysis. We evaluated 45 patients with chronic sustained atrial fibrillation (33 men and 12 women, age range 63 +/- 2 years) compared with 45 sex- and age- (+/- 2 years) matched nonhospitalized subjects with sinus rhythm. There were no significant differences between the two groups in the incidence of risk factors for stroke except for ischemic heart disease or in echocardiographic parameters. Plasma levels of NOx measured using the Greiss reagent (mean [interquartile range]: 15.6 [9.5 to 25.7] versus 24.1 [14.2 to 40.8] mumol/L, n = 45) and the platelet cGMP levels (0.33 [0.16 to 0.67] versus 0.63 [0.31 to 1.29] pmol/10(9) platelets, n = 9) were significantly (P < .05) lower in the patients with atrial fibrillation than in the control subjects. Plasma levels of D-dimer, beta thromboglobulin, and fibrinogen were significantly (P < .05) higher in the patients with atrial fibrillation. The two groups did not differ as to the plasma levels of tissue plasminogen activator or plasminogen activator inhibitor-1. Our findings suggest that a decrease in plasma NO levels may account for the hemostatic abnormalities observed in patients with atrial fibrillation. PMID- 9409312 TI - Shear-stress-induced detachment of blood platelets from various surfaces. AB - Platelet accumulation is the result of platelet adhesion and detachment. This study describes platelet detachment from fibronectin, laminin, fibrinogen, von Willebrand Factor (vWF), endothelial cell matrix (ECM), and collagen type III. Platelets adhered after 5 minutes' perfusion of anticoagulated whole blood at different shear rates were subjected to a brief flush of 1 minute with HEPES buffered saline at varying shear stress. Platelets adhering to fibronectin and laminin were most easily detached. Fibrinogen and vWF had an intermediate position. Almost no detachment occurred from ECM and collagen type III. Dendritic platelets were removed more easily than spread platelets. When the shear rate at which adhesion had occurred was raised, platelet detachment decreased strongly. When the time period between adhesion and detachment was increased, platelet detachment also decreased. From these results, we conclude that detachment is determined initially by the shear rate at which platelets adhere, then by the time they are allowed to settle, then by the nature of the surface, and then by the degree of spreading. The shear optimum for a given adhesive protein is not determined by the detachment. PMID- 9409311 TI - Role of angiotensin II and bradykinin on aortic collagen following converting enzyme inhibition in spontaneously hypertensive rats. AB - We previously showed that chronic angiotensin-converting enzyme (ACE) inhibition prevented the increase in aortic collagen in spontaneously hypertensive rats (SHRs) independently of blood pressure reduction. The aim of the present study was to determine whether the effects of ACE inhibition on aortic fibrosis were due to inhibition of angiotensin II formation, preservation of bradykinin, or a combination of both. Four week-old SHRs were treated for 4 months with the ACE inhibitor quinapril, quinapril with the bradykinin B2 receptor antagonist Hoe 140, or the angiotensin II AT1 receptor antagonist CI996. Control SHR and Wistar Kyoto (WKY) rats received a placebo for the same period of time. At the end of the treatment, as compared to conscious SHR and WKY controls, quinapril completely prevented the development of hypertension, whereas quinapril-Hoe 140 and the AT1 receptor antagonist produced only a partial reduction of blood pressure. In relation with blood pressure changes, aortic hypertrophy was significantly prevented by quinapril but not by quinapril-Hoe 140 or CI996. In contrast, aortic collagen accumulation was completely prevented by all three treatments. The study provides evidence that in young live SHRs, the prevention of aortic collagen accumulation is independent of blood pressure changes and bradykinin preservation and involves exclusively angiotensin II inhibition through AT1 receptors. PMID- 9409313 TI - Role of hemostatic risk factors for restenosis in peripheral arterial occlusive disease after transluminal angioplasty. AB - In a prospective study, the role of various hemostatic factors known to be associated with thrombotic risk was investigated in 71 patients with peripheral arterial occlusive disease (PAOD, stages II through IV, Fontaine; aged 68 +/- 13 years). Laboratory investigations were done before; 1, 24, and 48 hours after; and 3 and 6 months after percutaneous transluminal angioplasty (PTA). Thirty of 71 (42.3%) patients developed restenosis (> 50% reduction of the lumen diameter) at the site of PTA within 6 months, verified by color-coded duplex sonography. Significantly increased levels of thrombin-antithrombin III complexes (P < .01), prothrombin fragments 1 + 2 (P < .01), and D-dimers (P < .01) were found 1 hour, as well as 24 to 48 hours, after PTA. Fibrinogen (P < .01) and von Willebrand factor (P < .01) were significantly higher 48 hours after PTA. Restenotic patients as a whole had higher plasma fibrinogen (3.46 +/- 1.12 versus 2.95 +/- 0.62 g/L, P < .01) and C-reactive protein (25.4 +/- 46.7 versus 7.9 +/- 6.9 mg/L, P < .05) at baseline, as well as higher fibrinogen (P < .05) and prothrombin fragments 1 + 2 (P < .01) during months 3 to 6 after PTA. There was a nonsignificant tendency for higher values of von Willebrand factor (206 +/- 98% versus 184 +/- 100%, P = .2) at baseline in patients with restenosis, whereas tissue plasminogen activator, plasminogen activator inhibitor, coagulation screening tests, blood cell counts, and serum lipids showed no significant difference between the two groups. The relative risk for developing restenosis within 6 months while having high fibrinogen (> 2.8 g/L) or C-reactive protein at baseline was 2.80 (95% CI: 1.30-6.02, P < .01) and 1.96 (95% CI: 1.07-3.58, P < .05), respectively. Patients with critical limb ischemia (stage III/IV, Fontaine) had significantly higher fibrinogen and von Willebrand factor at repeated points of time, as well as significantly higher C-reactive protein and lower creatinine clearance at entry. In the logistic regression risk factor analysis, baseline plasma fibrinogen, C-reactive protein concentration, and the severity of the arterial disease were significantly predictive of restenosis. Our results indicate that high procoagulant factors and persistent thrombin generation of the hemostatic system might promote restenosis, particularly in patients with extended atherosclerosis. This finding suggests that new treatment strategies should be taken under consideration for patients with PAOD and PTA. PMID- 9409314 TI - Genotype-specific transcriptional regulation of PAI-1 expression by hypertriglyceridemic VLDL and Lp(a) in cultured human endothelial cells. AB - The hypothesized relationships between plasminogen activator inhibitor (PAI-1) genotypes, PAI-1 levels, and their potential regulation by hypertriglyceridemic (HTG) very low density lipoprotein (VLDL) and lipoprotein(a) [Lp(a)] was examined in a PAI-1 genotyped human umbilical vein endothelial cell (HUVEC) culture model system. Individual human umbilical veins were used to obtain cultured ECs and were genotyped for PAI-1 by using the HindIII restriction fragment length polymorphism (RFLP) as a marker for genetic variation. Digested genomic DNA, examined by Southern blot analysis and probed with an [alpha-32P]dCTP-labeled 2.2 kb PAI-1 cDNA, yielded three RFLPs designated 1/1 (22-kb band only), 1/2 (22-plus 18-kb bands), and 2/2 (18-kb band only). Individual PAI-1 genotyped HUVEC cultures were incubated in the absence or presence of HTG-VLDL (0 to 50 micrograms/mL) or Lp(a) (0 to 50 micrograms/mL) at 37 degrees C for various times (4 to 24 hours), followed by analyses of PAI-1 antigen (by ELISA) and mRNA (by ribonuclease protection assay) levels, EC surface-localized plasmin generation assays, and nuclear run-on transcription assays. Secreted PAI-1 antigen levels were increased approximately 2- to 3-fold by HTG-VLDL and approximately 1.6 to 2 fold by Lp(a); mRNA levels were increased approximately 3- to 4.5-fold by HTG VLDL and approximately 2.5- to 3.2-fold by Lp(a) compared with medium-incubated controls, primarily in the 2/2 PAI-1 genotype HUVEC cultures. Increases in PAI-1 mRNA induced by HTG-VLDL or Lp(a) could be abolished by coincubation with actinomycin D (2 x 10(-6) mol/mL) or puromycin (1 microgram/mL). In addition, nuclear transcription run-on assays typically demonstrated that HTG-VLDL increased PAI-1 gene transcription rates by approximately 5- to 6-fold and approximately 4- to 5-fold, respectively, primarily in the 2/2 PAI-1 genotype HUVEC cultures compared with 1/1 PAI-1 genotype HUVEC cultures or medium incubated controls. The positive control interleukin-1 increased both 2/2 and 1/1 PAI-1 mRNA levels by approximately 5- to 6-fold. Increased PAI-1 antigen and mRNA expression were associated with a concomitant 50% to 60% decrease in plasmin generation. These combined results demonstrate the genotype-specific regulation of PAI-1 expression by HTG-VLDL and Lp(a) and further indicate that these risk factor-associated components regulate PAI-1 gene expression at the transcriptional level in cultured HUVECs. Results from these studies further suggest that individuals with this responsive 2/2 PAI-1 genotype may reflect the additional inherent potential for later HTG-VLDL- or Lp(a)-induced fibrinolytic dysfunction, resulting in the early initiation of thrombosis, atherogenesis, and coronary artery disease. PMID- 9409315 TI - Continued thromboxane A2 formation despite administration of a platelet glycoprotein IIb/IIIa antagonist in patients undergoing coronary angioplasty. AB - Experimental data suggest that formation of thromboxane A2 may be suppressed during administration of a glycoprotein IIb/IIIa antagonist. We determined the dose of one such compound, fradafiban, required to provide > 80% occupancy of the platelet glycoprotein IIb/IIIa and examined its effects on thromboxane A2 formation in patients undergoing PTCA. The dose response to fradafiban and additional effects of aspirin were explored initially in patients with stable coronary artery disease. Fradafiban induced a dose-dependent inhibition of platelet aggregation that correlated with fibrinogen receptor occupancy and plasma drug concentration. Addition of aspirin 300 mg had no effect on these parameters. At the highest dose, mean fibrinogen receptor occupancy was 89.7 +/- 1.2% (n = 3) at 4 hours and platelet aggregation had decreased by 93.4 +/- 2.7%. Eighteen patients undergoing coronary angioplasty were randomized to receive either aspirin 330 mg or that dose of fradafiban producing > 80% fibrinogen receptor occupancy. Platelet aggregation was suppressed throughout the infusion of fradafiban to a greater extent than with aspirin. However, there was a marked increase in urinary excretion of 11-dehydrothromboxane B2 in patients treated with fradafiban: from 1973 +/- 889 to a peak of 9760 +/- 3509 pg/mg creatinine (P = .0046). Despite this evidence of continued platelet activation in vivo, there were no cases of coronary thrombosis. In conclusion, fradafiban suppresses platelet aggregation and may be a useful alternative to aspirin in the prevention of thrombotic events in patients undergoing PTCA. However, there is continued formation of thromboxane A2, which may continue to exert its effects as a potent vasoconstrictor and vascular smooth muscle mitogen. PMID- 9409316 TI - In vivo formation of 8-Epi-prostaglandin F2 alpha is increased in hypercholesterolemia. AB - F2-isoprostanes are bioactive prostaglandin (PG)-like compounds that are produced from arachidonic acid through a nonenzymatic process of lipid peroxidation catalyzed by oxygen free-radicals. 8-Epi-PGF2 alpha may amplify the platelet response to agonists, circulates in plasma, and is excreted in urine. We examined the hypothesis that the formation of 8-epi-PGF2 alpha is altered in patients with hypercholesterolemia and contributes to platelet activation in this setting. Urine samples were obtained from 40 hypercholesterolemic patients and 40 age- and sex-matched control subjects for measurement of immunoreactive 8-epi-PGF2 alpha. Urinary excretion of 11-dehydro-thromboxane (TX) B2, a major metabolite of TXA2, was measured as an in vivo index of platelet activation. Low-dose aspirin, indobufen, and vitamin E were used to investigate the mechanism of formation and effects of 8-epi-PGF2 alpha on platelet activation. Urinary 8-epi-PGF2 alpha was significantly (P = .0001) higher in hypercholesterolemic patients than in control subjects: 473 +/- 305 versus 205 +/- 95 pg/mg creatinine. Its rate of excretion was inversely related to the vitamin E content of LDL and showed a positive correlation with urinary 11-dehydro-TXB2. Urinary 8-epi-PGF2 alpha was unchanged after 2-week dosing with aspirin and indobufen despite complete suppression of TX metabolite excretion. Vitamin E supplementation was associated with dose dependent reductions in both urinary 8-epi-PGF2 alpha and 11-dehydro-TXB2 by 34% to 36% and 47% to 58% at 100 and 600 mg daily, respectively. We conclude that the in vivo formation of the F2-isoprostane 8-epi-PGF2 alpha is enhanced in the vast majority of patients with hypercholesterolemia. This provides an aspirin insensitive mechanism possibly linking lipid peroxidation to amplification of platelet activation in the setting of hypercholesterolemia. Dose-dependent suppression of enhanced 8-epi-PGF2 alpha formation by vitamin E supplementation may contribute to the beneficial effects of antioxidant treatment. PMID- 9409317 TI - Prostaglandin F2-like compounds, F2-isoprostanes, are present in increased amounts in human atherosclerotic lesions. AB - Oxidative modification of LDL is believed to play a major role in atherogenesis. As major lipid peroxidation products oxygenated linoleic acid derivatives and oxysterols have been described in human atherosclerotic lesions. Here we report that human lesions contain isoprostanes as peroxidation products of arachidonic acid at a level of 27.1 +/- 21.2 pg/mg wet weight (n = 10), which corresponds to 75.9 +/- 59.3 pg/mg dry weight, n contrast, human umbilical veins (n = 10), which were used as nonatherosclerotic control vessels, contain much smaller amounts of isoprostanes (1.4 +/- 0.7 pg/mg wet weight, which corresponds to 11.7 +/- 6.2 pg/mg dry weight), and there are significant differences between the two types of vessels. As major products of linoleic acid oxidation, racemic hydroxy linoleate isomers were detected in the lesional ester lipids. In human lesions, the hydroxy linoleic acid/linoleic acid ratio was about 0.5%, a result indicating that 5 out of 1000 linoleate residues are present as hydroxylated derivatives. In umbilical veins, no hydroxy linoleic acid could be detected. These data show that human atherosclerotic lesions contain increased amounts of hydroxy linoleic acid isomers and isoprostanes when compared with nonatherosclerotic vessel wall and suggest a link between local lipid peroxidation and progression of atherosclerosis. For evaluation of the degree of lipid peroxidation, the determination of the hydroxy linoleic acid/linoleic acid ratio appears to be more suitable than the isoprostane content. PMID- 9409318 TI - Polymorphism of angiotensin converting enzyme gene is associated with circulating levels of plasminogen activator inhibitor-1. AB - The deletion (D) allele of the insertion/deletion (I/D) polymorphism of the angiotensin converting enzyme (ACE) gene is strongly associated with an increased level of circulating ACE. The ACE gene polymorphism may influence the production of angiotensin II (Ang II). It has been shown that Ang II modulates fibrinolysis, that is, Ang II increases plasminogen activator inhibitor-1 (PAI-1) mRNA and plasma PAI-1 levels in vitro and in vivo. Considered together, we tested the hypothesis that the deletion allele of the ACE gene might be associated with increased levels of PAI-1. We related the ACE genotype to PAI-1 antigen levels in 603 men and 221 women attending a routine health screening. As a whole, the plasma PAI-1 level was not strongly associated with ACE genotype. Since the PAI-1 level was significantly influenced by well-known risk factors for coronary artery disease (CAD), we further analyzed the data after excluding subjects with major cardiovascular risk factors. In low-risk male subjects, the DD genotype had significantly higher levels of plasma PAI-1 (DD: 20.3 +/- 2.2; DI: 13.9 +/- 1.1; II: 13.6 +/- 1.3 ng/mL, P = .010 by ANOVA). In low-risk female subjects, the DD genotype showed a tendency to a high level of plasma PAI-1 without statistical significance. When analysis was restricted to postmenopausal women (age > or = 55 or FSH > or = 35 ng/mL), the DD genotype showed a significantly higher level of PAI-1 than subjects with the DI and II genotypes (27.7 +/- 6.2 versus 15.6 +/- 1.8 ng/mL, P = .028). The DD polymorphism of the ACE gene is associated with high PAI-1 levels in male and possibly in postmenopausal female subjects who have lower conventional cardiovascular risk factors. These results suggest that the increased ACE activity caused by DD polymorphism may play an important role in elevating the level of plasma PAI-1. Our data support the notion that the genetic variation of ACE contributes to the balance of the fibrinolytic pathway. PMID- 9409319 TI - Evidence that human Fc gamma receptor IIA (CD32) subtypes are not receptors for oxidized LDL. AB - Several lines of evidence suggest that clearance of oxidized LDL (oxLDL) immune complexes by macrophage IgG Fc receptors (Fc gamma Rs) plays a role in atherogenesis. Ox-LDL may also be cleared directly by Fc gamma Rs, as shown for murine Fc gamma RII-B2. In humans, the homologous Fc gamma R is Fc gamma RIIA (CD32), which is abundantly expressed on monocytes and macrophages and shares 60% sequence identity with murine Fc gamma RII-B2. As murine Fc gamma RII-B2 and human Fc gamma RIIA also share similar IgG ligand-binding properties, the purpose of this study was to test the hypothesis that human CD32 is a receptor for oxLDL. For these studies we used transfected Chinese hamster ovary (CHO) cells, monocytes, and cell lines that functionally express either of two Fc gamma RIIA subtypes (R131 or H131) and assayed binding or degradation of several preparations of oxLDL. The integrity of all oxLDL preparations was checked by studying their ability to react with CHO cells expressing human type I scavenger receptors and by other characteristics of lipoprotein oxidation. Although we showed that each preparation of oxLDL could recognize class A or class B scavenger receptors, we did not detect any differences in the binding or degradation of any type of oxLDL preparation among control versus CHO cell transfectants. Using monocytes that express Fc gamma RIIA and CD36, we showed that the binding of oxLDL was inhibited by antibodies to CD36, but not by Fc gamma RIIA antibodies. Thus, the data do not support the hypothesis that human Fc gamma RIIA is by itself a receptor for oxLDL. We conclude that human CD32 can mediate uptake of lipoprotein immune complexes, but does not mediate uptake of oxLDL in the absence of anti-oxLDL antibodies. OxLDL may interact with human mononuclear phagocytes directly via other types of receptors, such as class A and class B scavenger receptors or CD68. PMID- 9409320 TI - Tetradecylthioacetic acid inhibits the oxidative modification of low density lipoprotein and 8-hydroxydeoxyguanosine formation in vitro. AB - Oxidative modification of low-density lipoprotein (LDL) is thought to play a key role in the formation of foam cells and in initiation and progression of atherosclerotic plaque. The hypolipidemic 3-thia fatty acids contain a sulfur atom and might therefore possess reducing (antioxidant) properties. Consequently, the effects of 3-thia fatty acids on the susceptibility of LDL particles to undergo oxidative modification in vitro were studied. Tetradecylthioacetic acid (TTA), incorporated into the LDL particle and increased the lag time of copper ion induced LDL oxidation in a dose-dependent manner, 80 mumol/L TTA reduced the generation of lipid peroxides during copper ion induced LDL oxidation (for 2 hours) by 100%, 2,2'-azobis-(2,4-dimethylvaleronitrile) induced LDL oxidation by 64%, and 2,2'-azobis-(2-amidinopropane hydrochloride) induced LDL oxidation (for 6 hours) by 21%. The electrophoretic mobility of the oxidized LDL was reduced by TTA in both copper ion and azo-compounds initiated oxidation. This fatty acid analogue was effectively able to reduce in a dose dependent manner the formation of 8-hydroxydeoxyguanosine from 2-deoxyguanosine with ascorbic acid as the radical producer. TTA bound copper(II) ions and did not reduce copper(II) to copper(I). It failed to scavenge the 1,1-diphenyl-2-picrylhydrazyl radicals. The results suggest that the modification of LDL in the lipid and protein moieties can be significantly reduced by TTA. This acid may exert its antioxidant effect partially through metal ion binding and through free radical scavenging. PMID- 9409321 TI - Familial resemblance of plasma lipids, lipoproteins and postheparin lipoprotein and hepatic lipases in the HERITAGE Family Study. AB - The familial aggregation of lipids and lipoproteins and plasma postheparin triglyceride lipases was investigated in 86 Caucasian families participating in the HERITAGE Family study, a study investigating the role of genetic factors in the adaptation to exercise training and its relationships with cardiovascular disease risk factors. Accordingly, sedentary subjects were recruited, tested for a battery of measurements, exercise trained for 20 weeks, and were re-measured. The present report includes plasma levels of total cholesterol, high-density lipoprotein (HDL) cholesterol, low-density lipoprotein (LDL) cholesterol and triglycerides, and postheparin plasma lipoprotein lipase (LPL) and hepatic lipase (HL) activities measured in 437 sedentary individuals (171 parents and 266 adult offspring) before training. Significant familial resemblance was observed for all the age-adjusted phenotypes. The pattern of familial correlations reveals no spouse correlations but significant parent-offspring and sibling correlations for total cholesterol, HDL-cholesterol and LDL-cholesterol with heritability (h2) estimates of 62%, 83%, and 50%, respectively. For plasma triglyceride concentrations (h2 = 55%) and HL activity (h2 = 40%), significant spouse correlations were found in addition to parent-offspring and sibling correlations, suggesting that common familial environment in addition to genetic factors contribute to the familial resemblance. For plasma LPL activity, there was no spouse correlation, but sex differences were found in the familial correlations with higher heritabilities in female pairs (h2 = 76%) compared to male pairs (h2 = 30%) and opposite-sex pairs (h2 = 44%). These results confirm the findings of previous family studies showing that genetic factors are major determinants of the familial resemblance in plasma lipids and lipoproteins and suggest the presence of sex differences in the heritability of postheparin LPL activity. PMID- 9409322 TI - Cross-trait familial resemblance for body fat and blood lipids: familial correlations in the Quebec Family Study. AB - In an attempt to better understand the genetic basis of the metabolic syndrome, we have undertaken a series of studies on the familial aggregation in the clustering of the coronary heart disease risk factors which characterize this syndrome. In the present study, the hypothesis of shared genetic (pleiotropy) and/or environmental factors between body fat and blood lipids is investigated by examining cross-trait (eg, father's body fat with his son's blood lipid) familial resemblance between 4 indicators of body fat (body mass index [BMI], sum of 6 skin folds [SF6]) and fat distribution (the ratio of the trunk to extremity skin folds adjusted [TER-sf] and unadjusted [TER] for SF6), and 5 blood lipid variables (total cholesterol [CH], triglycerides [TG], cholesterol associated with high-density lipoproteins [HDL], the CH/ HDL ratio and the difference between CH and HDL [CH-HDL]) measured in 1239 individuals from 309 families participating in the Quebec Family Study. A bivariate correlation model was used to obtain maximum likelihood estimates of cross-trait spouse, parent-offspring, and sibling correlations after adjustment of body fat and lipid data for the effects of age, separately in the four sex-by-generation groups. Likelihood ratio tests revealed the presence of significant (P < .05) cross-trait resemblance between body fat (BMI and SF6) and all lipid traits except CH and also between fat distribution (TER and TER-sf) and CH/HDL and CH-HDL. Only sibling cross-trait correlations were significant for all body fat-lipid pairs of measures, with bivariate familiality estimates (i.e., shared genetic and/or environmental factors) ranging from 8% to 40%. Although the hypothesis of genetic pleiotropy cannot be ruled out from the pattern of cross-trait correlations found in the present study, we conclude that environmental factors shared within sibships are probably more important than common genes in determining the covariation between body fat and blood lipids. PMID- 9409323 TI - Identification of the Src family kinases, Lck and Fgr in platelets. Their tyrosine phosphorylation status and subcellular distribution compared with other Src family members. AB - We have identified the Src family members, Lck and Fgr in resting human and rodent platelets and compared their subcellular distributions and tyrosine phosphorylation status to those of the other Src family kinases to gain insights into the signal transduction pathways active in maintaining platelets in the circulation. Like Fyn, Lyn, and Yes, most of Fgr and Lck was detergent-insoluble in human and rat platelets. In comparison, Src showed higher detergent solubility than the Src-related kinases. Most all human platelet Src was detergent-soluble, while that of rodent platelets was present in all detergent fractions. We also compared the tyrosine-phosphorylation status of Lck and Fgr to other Src family members in resting platelets using immunoprecipitation and immunoblotting. All of these Src family members except Fgr exhibited substantial phosphotyrosine antibody labeling. The partitioning of these kinases, with the exception of Src, with the detergent-insoluble fraction, their tyrosine-phosphorylation status, and co-localization with endocytotic vesicles lead us to hypothesize that the Src family kinases are involved in signaling events that drive cytoskeletal reorganization and active endocytosis of plasma proteins by circulating platelets. PMID- 9409324 TI - Interaction between BTBR and C57BL/6J genomes produces an insulin resistance syndrome in (BTBR x C57BL/6J) F1 mice. AB - Insulin resistance is a common syndrome that often precedes the development of noninsulin-dependent diabetes mellitus (NIDDM). Both diet and genetic factors are associated with insulin resistance. BTBR and C57BL/6J (B6) mice have normal insulin responsiveness and normal fasting plasma insulin levels. However, a cross between these two strains yielded male offspring with severe insulin resistance. Surprisingly, on a basal diet (6.5% fat), the insulin resistance was not associated with fasting hyperinsulinemia. However, a 15% fat diet produced significant hyperinsulinemia in the male mice (twofold at 10 weeks; P < .05). At 10 weeks of age, visceral fat contributed approximately 4.3% of the total body weight in the males versus 1.8% in females. In the males, levels of plasma triacylglycerol and total cholesterol increased 40% and 30%, respectively, compared to females. Plasma free fatty acid concentrations were unchanged. Oral glucose tolerance tests revealed significant levels of hyperglycemia and hyperinsulinemia 15 to 90 minutes after oral glucose administration in the male mice. This was particularly dramatic in males on a 15% fat diet. Glucose transport was examined in skeletal muscles in (BTBR x B6)F1 mice. In the nonhyperinsulinemic animals (females), insulin stimulated 2-deoxyglucose transport 3.5-fold in the soleus and 2.8-fold in the extensor digitorum longus muscles. By contrast, glucose transport was not stimulated in the hyperinsulinemic male mice. Hypoxia stimulates glucose transport through an insulin-independent mechanism. This is known to involve the translocation of GLUT4 from an intracellular pool to the plasma membrane. In the insulin-resistant male mice, hypoxia induced glucose transport as effectively as it did in the insulin-responsive mice. Thus, defective glucose transport in the (BTBR x B6)F1 mice is specific for insulin-stimulated glucose transport. This is similar to what has been observed in muscles taken from obese NIDDM patients. These animals represent an excellent genetic model for studying insulin resistance and investigating the transition from insulin resistance in the absence of hyperinsulinemia to insulin resistance with hyperinsulinemia. PMID- 9409325 TI - Changes in arterial expression of fibrinolytic system proteins in atherogenesis. AB - Plasminogen activators (PAs) and their inhibitor, plasminogen activator inhibitor type-1 (PAI-1), have been implicated in modulation of luminal fibrinolysis and mural proteolysis contributing to atherogenesis. Expression of PAs/PAI-1 (normalized to extracted tissue protein) was delineated by assays of conditioned media and of extracts from walls of human arterial segments in culture. Arterial specimens (n = 39 from 26 subjects) were divided into four groups: normal (n = 14), fatty streak (n = 6), moderate atherosclerosis (mural thickening with < 70% lumen obstruction, n = 5), and severe atherosclerosis (mural thickening with > 70% lumen obstruction, n = 14). Paired samples from the same individual comprising a normal arterial segment and an atherosclerotic segment were evaluated also. A fourfold molar excess in PAI-1:t-PA was seen in conditioned media from samples with any evidence of atherosclerosis compared with normal specimens (normal 21 +/- 4, diseased 82 +/- 21, P < or = .05). Compared with normal pairs, the tissue content of PAI-1 (ng) was increased in fatty streak lesions (n = 3, normal 35 +/- 12, fatty streak 50 +/- 8, P < or = .05); stable to decreased in moderate atherosclerosis (n = 3, normal 34 +/- 3, moderate 22 +/- 7, P = .16); and increased in severe atherosclerosis (n = 6, normal 48 +/- 9, severe 85 +/- 19, P < or = .05). The tissue content of PAs (ng), though not increased in fatty streak lesions, was elevated in moderately and severely atherosclerotic segments (normal 0.7 +/- 0.2, moderate 1.6 +/- 0.1; normal 0.8 +/- 0.3, severe 2.1 +/- 0.3, P < or = .05 for each comparison). Atherogenesis is associated with decreased luminal fibrinolytic capacity that may exacerbate thrombosis. Decreased mural proteolysis in early atherogenesis may exacerbate matrix accumulation. Increased mural proteolysis later is associated with, and may potentiate, smooth muscle cell migration and proliferation. PMID- 9409326 TI - Insulin resistance as an independent risk factor for carotid artery wall intima media thickening in vasospastic angina. AB - Studies have shown the presence of insulin resistance together with compensatory hyperinsulinemia in vasospastic angina as well as obstructive coronary artery disease. There is growing evidence that the development of coronary atherosclerosis may be closely related to systemic atherosclerosis as well as coronary spasm. However, no information is available about the possible relationship between insulin resistance and the existence of carotid atherosclerosis in vasospastic angina without segmental stenosis or luminal irregularities in coronary angiograms. To evaluate the independent effect of insulin resistance on carotid intima media thickening, we performed insulin sensitivity tests (steady-state plasma glucose method) on 40 patients with vasospastic angina and 24 control subjects with angiographically intact coronary arteries. Both oral glucose tolerance tests and lipid analyses were performed. Using B-mode ultrasonography, we assessed intima media thickness and plaque formation of common carotid arteries in these subjects. The steady-state plasma glucose level in the vasospastic angina group was about twofold higher than that of the control group, confirming the presence of insulin resistance in patients with vasospastic angina. The patients with vasospastic angina showed a significant increase in the average intima media thickness of the carotid wall and frequency of plaque formation, although they were comparable to the control subjects in risk factors other than insulin resistance. The intima media thickness was correlated with age (r = .62, P < .001), 2-hour insulin area (r = .45, P < .01), and steady-state plasma glucose level (r = .68, P < .0001) in patients with vasospastic angina. Similar correlations were observed in the control subjects. Multiple regression analyses of data indicated that 67% of the variation in the intima media thickness could be accounted for by age, steady state plasma glucose level, and cigarette-years in vasospastic angina. In addition, differences in IMT were independently related to vasospastic angina. These results suggest that insulin resistance in association with compensatory hyperinsulinemia may be an important pathogenic factor for the development of coronary artery spasms and systemic early atherosclerosis. PMID- 9409327 TI - alpha IIb beta 3 redistribution triggered by receptor cross-linking. AB - Fibrinogen binding to alpha IIb beta 3 on adherent, spread platelets triggers active, cytoskeletally-directed redistribution of fibrinogen/alpha IIb beta 3 complexes on the platelet surface. Gold-conjugated fibrinogen, unlabeled, soluble fibrinogen, and individual fibrinogen molecules have been demonstrated to trigger receptor redistribution. Here we examine the respective roles of receptor cross linking and ligand occupancy of receptors in initiating this movement. Monovalent, alpha IIb beta 3-binding fibrinogen fragments RGDS and HHLGGAKQAGDV did not trigger receptor redistribution, suggesting that ligand binding to a single receptor is an insufficient stimulus. Binding of monoclonal antibodies 10E5, AP2, and AP3 to the receptor did not trigger receptor movement. However, cross-linking these receptor-bound monoclonal antibodies by polyclonal anti-mouse IgG or by conjugation of the anti-receptor antibody to large colloidal gold particles triggered receptor redistribution identical in rate, pattern, and final distribution to that previously seen with fibrinogen binding. We conclude that receptor cross-linking provides the signal for initiation of fibrinogen/alpha IIb beta 3 complex redistribution on platelet surfaces. PMID- 9409328 TI - Hemostatic factors as predictors of ischemic heart disease and stroke in the Edinburgh Artery Study. AB - Plasma fibrinogen is a consistent predictor of ischemic heart disease (IHD) in prospective studies, but there are fewer data relating other hemostatic variables to IHD and also to stroke. We therefore studied the relationships of plasma fibrinogen, von Willebrand factor antigen, tissue plasminogen activator (TPA) antigen, factor VII, and fibrin D-dimer to incidence of IHD and stroke and determined whether any associations could be explained by conventional risk factors and baseline heart disease. In the Edinburgh Artery study, 1592 men and women aged 55 to 74 years, randomly sampled from the general population, were followed prospectively over 5 years to detect fatal and nonfatal IHD and stroke events. During the 5 years, 268 new vascular events were identified. Baseline plasma fibrinogen was independently related to risk of stroke in multivariate analysis that adjusted for cigarette smoking, LDL-cholesterol, systolic blood pressure, and preexisting IHD (relative risk [RR] 1.52, 95% confidence interval [CI] 1.17, 1.98). TPA antigen, and fibrin D-dimer were also independently associated with risk of stroke (RR 1.69,95% CI 1.22,2.35 and RR 1.96, 95% CI 1.12,3.41, respectively). Significant relationships were found between TPA antigen and myocardial infarction (P < or = .05). In older men and women, increased coagulation activity and disturbed fibrinolysis are predictors of future vascular events (both IHD and stroke). PMID- 9409329 TI - Interaction of diet and genes in atherogenesis. Report of an NHLBI working group. AB - Recent advances in genetics and information emerging from the Human Genome Project make it feasible to examine the importance of dietary-genetic interactions in the development of atherosclerosis. In the opinion of the Working Group, three approaches are necessary to examine this concern. The first approach utilizes animal models to map and identify candidate genes involved in dietary responsiveness and atherogenesis. The second approach involves the evaluation of these genes in specific physiological processes involved in dietary responsiveness and atherogenesis. Finally, the third approach is to extend the studies performed in animal models to human populations using linkage or association studies. PMID- 9409330 TI - Neonatal diagnosis of familial hypercholesterolemia in newborns born to a parent with a molecularly defined heterozygous familial hypercholesterolemia. AB - This study was designed to compare blood lipid levels in newborn individuals with molecularly defined heterozygous familial hypercholesterolemia [FH] to those in non-affected babies and to clarify the value of lipid determinations in assessment of diagnosis of FH at birth and 1 year of age. Twenty-five babies were born to 21 parents with DNA-documented heterozygous FH. Analysis of their cord blood samples revealed 11 newborns with the FH-North Karelia [FH-NK] mutation, 3 newborns with the FH-Helsinki [FH-HKI] mutation, and 11 nonaffected newborns. Cord serum total [TC] and LDL cholesterol [LDL-C] levels (mean +/- SD) in affected newborns (2.60 +/- 0.70 and 1.77 +/- 0.56, respectively) were significantly (P < .001) higher than those in nonaffected ones (1.54 +/- 0.23 and 0.78 +/- 0.15, respectively) and another cohort of 30 randomly selected control samples from apparently healthy newborns (1.84 +/- 0.46 and 1.03 +/- 0.30, respectively). However, there was overlapping of individual lipid levels in these three groups precluding the use of TC or LDL-C determinations in neonatal diagnosis of FH. In contrast, 1 year follow-up samples from 10 affected and 7 nonaffected individuals, as well as additional samples collected from another group of 8 affected and 9 nonaffected individuals, indicated that serum cholesterol levels showed much greater increment in children with FH. Thus, at the age of 1 year the mean serum TC and LDL-C levels in the affected infants (8.38 +/- 1.18 and 7.02 +/- 1.07, respectively) were much higher (P < .001) than the corresponding levels (4.40 +/- 0.66 and 2.89 +/- 0.68, respectively) in the nonaffected infants, and the individual ranges of TC and LDL-C levels were nonoverlapping in these two groups. Serum HDL cholesterol [HDL-C] levels in 1 year-old children with FH (0.95 +/- 0.14) were approximately 20% lower than those of their similar at birth. In conclusion, phenotypic expression of heterozygous FH, as defined by molecular analysis of genomic DNA, is evident in serum LDL-C (but not HDL-C) levels already at birth, but for diagnostic purposes blood lipid determinations carried out at the age of 1 year are highly superior to those performed at birth. PMID- 9409331 TI - Copper ions promote peroxidation of low density lipoprotein lipid by binding to histidine residues of apolipoprotein B100, but they are reduced at other sites on LDL. AB - Oxidized LDL is implicated in the pathogenesis of atherosclerosis. A widely studied model for oxidation of the lipid in LDL involves Cu2+. Recent studies suggest that Cu2+ may be reduced to Cu1+ by alpha-tocopherol to initiate LDL lipid peroxidation. LDL demonstrates binding sites for Cu2-, but the nature of these binding sites, as well their role in promoting Cu2+ reduction and lipid peroxidation, has not been established. In the current studies, we used diethylpyrocarbonate (DEPC) to modify the histidine residues of apolipoprotein B100, the major protein in LDL. First, we demonstrated that histidine residues were preferentially modified by DEPC under our experimental conditions. Then we monitored the kinetics of Cu(2+)-promoted oxidation of LDL and DEPC-modified LDL. In both cases, the progress curve of lipid peroxidation exhibited a lag phase and a propagation phase. However, when LDL was modified with DEPC, the length of the lag phase was prolonged whereas the rate of lipid peroxidation during the propagation phase was lower. Studies with LDL oxidized by 2,2'-azobis (2 amidinopropane) hydrochloride and phosphatidylcholine liposomes oxidized with hydroxyl radical established that DEPC was not acting simply as a nonspecific inhibitor of lipid peroxidation. DEPC treatment of LDL almost completely inhibited its ability to bind Cu2+. These observations suggest that peroxidation of the lipids in LDL can proceed with normal kinetics only when Cu2+ binds preferentially to sites on apolipoprotein B100 that contain histidine residues. We also compared the kinetics of Cu2+ reduction in the absence and presence of DEPC. There was no effect of DEPC modification on either the rate or extent of Cu2+ reduction by LDL. Therefore LDL is likely to contain a second class of binding sites for Cu2+ that does not involve histidine residues. Thus, LDL appears to contain at least two classes of Cu(2+)-binding sites: histidine containing sites, which are responsible in part for promoting lipid peroxidation during the propagation phase, and sites at which Cu2+ is reduced without binding to histidine. PMID- 9409332 TI - Demonstration of biphasic effects of docosahexaenoic acid on apolipoprotein B secretion in HepG2 cells. AB - Oleic acid (OA) stimulates apolipoprotein B (apoB) secretion from HepG2 cells by protecting the nascent protein from rapid intracellular degradation. In contrast, the n-3 fatty acids, docosahexaenoic acid (DHA) and eicosapentaenoic acid, have been shown to reduce apoB secretion by increasing its intracellular degradation in rat hepatocytes. We attempted to determine if OA and DHA have these opposite effects at the same point in the secretory pathway for apoB or if they act at different points in HepG2 cells. Unexpectedly, we found that when DHA (0.2 mmol/L) was incubated with HepG2 cells for 2 hours, it stimulated both triglyceride (TG) synthesis and apoB secretion significantly (the "stimulatory effect"). The stimulatory effect of DHA on apoB secretion was associated with decreased intracellular degradation of newly synthesized apoB. These acute effects of DHA on TG synthesis and apoB secretion paralleled those previously demonstrated with OA. After DHA was removed from the medium, however, both TG synthesis and apoB secretion rapidly decreased to a level that was significantly less than the control level (the "inhibitory effect"). At the same time, intracellular apoB degradation was significantly increased, and this degradation was efficiently prevented by proteasome inhibitors. Removal of DHA from the incubation resulted in inhibition of the incorporation of endogenous fatty acids into TG. In contrast, removal of OA from the media was not associated with any such inhibitory effect. The inhibitory effect of DHA on basal apoB secretion persisted at least 8 hours. These studies suggest that incubation of HepG2 cells with DHA has biphasic effects on TG synthesis and apoB secretion: an initial stimulation of TG synthesis is followed by inhibition of TG synthesis and increased apoB degradation. Although the stimulatory effect of DHA is apparent during short incubations of HepG2 cells, both effects would be expected to occur during long incubations, since fatty acid uptake by cells is rapid and efficient. Thus, long incubations of HepG2 cells with DHA could result in overall reduced apoB secretion compared with cells incubated in bovine serum albumin. If these findings are extrapolated to the in vivo situation, they can explain the ability of dietary n-3 fatty acids, which would be delivered to the liver intermittently, to reduce very low density lipoprotein secretion. PMID- 9409333 TI - Differential effects of extracellular Mg2+ on thrombin-induced and capacitative Ca2+ entry in human coronary arterial endothelial cells. AB - Receptor-mediated and capacitative Ca2+ entry are the primary Ca2+ entry pathways in endothelial cells (ECs). The mechanisms for Ca2+ entry via these pathways have not been fully elucidated. In this study, the effect of low and high external Mg2+ concentrations on these Ca2+ entry pathways was examined in human coronary arterial ECs. External Mg2+ concentration did not affect cytosolic free Mg2+ concentration. After exposure to thrombin in Ca(2+)-free medium, addition of Ca2+ to the medium caused a rise in cytosolic free Ca2+ concentration ([Ca2+]i), indicating thrombin-induced Ca2+ influx. Thrombin-induced Ca2+ influx was inhibited by not only low but also high external Mg2+ concentrations. After depletion of endoplasmic Ca2+ stores by thapsigargin, addition of Ca2+ to the medium induced an increase in [Ca2+]i, indicating capacitative Ca2+ entry. Capacitative entry was found to be accelerated by low external Mg2+ and inhibited by high external Mg2+ concentration. Results suggest that receptor-mediated Ca2+ influx requires external Mg2+ but is inhibited by increased external Mg2+ concentrations and that capacitative Ca2+ entry is reduced by external Mg2+ in human coronary arterial ECs. PMID- 9409335 TI - Estrogen replacement therapy and longitudinal decline in visual memory. A possible protective effect? AB - Estrogen replacement therapy (ERT) is increasingly recommended for postmenopausal women due to its beneficial effects on physical health in older women. Recent studies have suggested that ERT may have a protective effect on cognitive function and may reduce the risk of Alzheimer's disease. In the present study we test the hypothesis that ERT may have a protective effect on memory in nondemented women. Data on hormonal status and memory were examined in 288 postmenopausal women in the Baltimore Longitudinal Study of Aging. One hundred sixteen women who reported that they were receiving ERT during a cognitive assessment were compared with 172 women who had never received ERT. Women who were receiving ERT had fewer errors on the Benton Visual Retention Test (BVRT), a measure of short-term visual memory, visual perception, and constructional skills. Furthermore, ERT appeared to protect against age changes in BVRT performance in a subgroup of 18 women for whom BVRT data were available before and during treatment with ERT. These findings suggest that ERT may protect against memory decline in nondemented postmenopausal women and offer further support for a beneficial role of estrogen on cognitive function in aging women. PMID- 9409334 TI - Naratriptan is effective and well tolerated in the acute treatment of migraine. Results of a double-blind, placebo-controlled, crossover study. The Naratriptan S2WA3003 Study Group. AB - The efficacy and tolerability of naratriptan tablets (2.5 mg, 1 mg, and 0.25 mg) compared with placebo in the acute treatment of migraine were evaluated in a randomized, double-blind, four-period crossover study. Five hundred eighty-six assessable patients received naratriptan 2.5 mg, 595 received 1 mg, 591 received 0.25 mg, 602 received placebo. Headache relief (moderate or severe pain reduced to mild or none) 4 hours postdose was reported in 68% of patients after treatment with naratriptan 2.5 mg compared with 57% after 1 mg, 39% after 0.25 mg, and 33% after placebo (p < 0.001 naratriptan 2.5 mg and 1 mg versus placebo and 1 mg and 2.5 mg versus 0.25 mg). Headache relief was maintained 8, 12, and 24 hours postdose with no use of rescue medication or a second dose of study medication by significantly (p < 0.001) greater percentages of patients after treatment with naratriptan 2.5 mg or 1 mg compared with naratriptan 0.25 mg or placebo. Naratriptan was also more effective than placebo in reducing clinical disability and the incidences of nausea, photophobia, and phonophobia. The overall incidence of adverse events and the incidences of specific adverse events did not differ in the naratriptan groups compared with placebo. No clinically relevant changes in ECG, blood pressure, or laboratory findings were reported. These data demonstrate that naratriptan is effective and well tolerated for the acute treatment of migraine. The 2.5-mg dose was associated with superior efficacy, whereas its adverse event profile was similar to that of placebo. PMID- 9409336 TI - Lifetime risk of dementia and Alzheimer's disease. The impact of mortality on risk estimates in the Framingham Study. AB - We estimated the remaining lifetime risks of developing Alzheimer's disease (AD) and dementia from all causes, based on data from longitudinal population studies. The risk of developing AD during one's lifetime depends on both disease incidence and life expectancy. Conventional estimates of cumulative incidence overestimate the risk when there is a substantial probability of mortality due to competing causes. A total of 2,611 cognitively intact subjects (1,061 men, 1,550 women; mean age, 66 +/- 7 years) were prospectively evaluated for the development of AD or other dementia. A modified survival analysis was used to estimate both cumulative incidence and the sex-specific remaining lifetime risk estimates for quinquennial age groups above age 65 years. Over a 20-year follow-up period, 198 subjects developed dementia (120 with AD). The remaining lifetime risk of AD or other dementia depended on sex, being higher in women, but varied little with age between 65 and 80 years. In a 65-year-old man, the remaining lifetime risk of AD was 6.3% (95% CI, 3.9 to 8.7) and the remaining lifetime risk of developing any dementing illness was 10.9% (95% CI, 8.0 to 13.8); corresponding risks for a 65 year-old woman were 12% (95% CI, 9.2 to 14.8) and 19% (95% CI, 17.2 to 22.5). The cumulative incidence between age 65 and 100 years was much higher: for AD, 25.5% in men and 28.1% in women; for dementia, 32.8% in men and 45% in women. The actual remaining lifetime risk of AD or dementia varies with age, sex, and life expectancy and is lower than the hypothetical risk estimated by a cumulative incidence in the same population. PMID- 9409337 TI - Quantitative MR volumetry in Alzheimer's disease. Topographic markers and the effects of sex and education. AB - We determined topographic selectivity and diagnostic utility of brain atrophy in probable Alzheimer's disease (AD) and correlations with demographic factors such as age, sex, and education. Computerized imaging analysis techniques were applied to MR images in 32 patients with probable AD and 20 age- and sex-matched normal control subjects using tissue segmentation and three-dimensional surface rendering to obtain individualized lobar volumes, corrected for head size by a residualization technique. Group differences emerged in gray and white matter compartments particularly in parietal and temporal lobes. Logistic regression demonstrated that larger parietal and temporal ventricular CSF compartments and smaller temporal gray matter predicted AD group membership with an area under the receiver operating characteristic curve of 0.92. On multiple regression analysis using age, sex, education, duration, and severity of cognitive decline to predict regional atrophy in the AD subjects, sex consistently entered the model for the frontal, temporal, and parietal ventricular compartments. In the parietal region, for example, sex accounted for 27% of the variance in the parietal CSF compartment and years of education accounted for an additional 15%, with women showing less ventricular enlargement and individuals with more years of education showing more ventricular enlargement in this region. Topographic selectivity of atrophic changes can be detected using quantitative volumetry and can differentiate AD from normal aging. Differential effects of sex and years of education can also be detected by these methods. Quantification of tissue volumes in vulnerable regions offers the potential for monitoring longitudinal change in response to treatment. PMID- 9409338 TI - Changes of hippocampal N-acetyl aspartate and volume in Alzheimer's disease. A proton MR spectroscopic imaging and MRI study. AB - Hippocampal atrophy detected by MRI is a prominent feature of early Alzheimer's disease (AD), but it is likely that MRI underestimates the degree of hippocampal neuron loss, because reactive gliosis attenuates atrophy. We tested the hypothesis that hippocampal N-acetyl aspartate (NAA: a neuronal marker) and volume used together provide greater discrimination between AD and normal elderly than does either measure alone. We used proton MR spectroscopic imaging (1H MRSI) and tissue segmented and volumetric MR images to measure atrophy-corrected hippocampal NAA and volumes in 12 AD patients (mild to moderate severity) and 17 control subjects of comparable age. In AD, atrophy-corrected NAA from the hippocampal region was reduced by 15.5% on the right and 16.2% on the left (both p < 0.003), and hippocampal volumes were smaller by 20.1% (p < 0.003) on the right and 21.8% (p < 0.001) on the left when compared with control subjects. The NAA reductions and volume losses made independent contributions to the discrimination of AD patients from control subjects. When used separately, neither hippocampal NAA nor volume achieved to classify correctly AD patients better than 80%. When used together, however, the two measures correctly classified 90% of AD patients and 94% of control subjects. In conclusion, hippocampal NAA measured by 1H MRSI combined with quantitative measurements of hippocampal atrophy by MRI may improve diagnosis of AD. PMID- 9409339 TI - An alphabetical 'WORLD'. A new version of an old test. AB - The Mini-Mental State Examination (MMSE) is a standardized test used by neurologists to screen patients for impaired cognition. Despite its ease of use, one major limitation of the MMSE is a possible ceiling effect or a lower sensitivity in patients with advanced education. Patients (n = 97) undergoing diagnostic neuropsychological testing also completed one subtest of the MMSE, the spelling of "world." In addition to its forward and backward spelling, patients were asked to reorder these letters in alphabetical sequence. Our preliminary data indicate that our modified WORLD test is a rapid and simple test to identify patients with cognitive impairment. When measured against the diagnosis of dementia as determined by neuropsychological testing, the modified WORLD test has a sensitivity of 85%, a specificity of 88%, and a positive predictability value of 95%. Other variables examined include patient age, sex, education, and cutoff scores on the Mattis Dementia Rating Scale. PMID- 9409340 TI - Normalization of neuronal metabolic dysfunction after surgery for temporal lobe epilepsy. Evidence from proton MR spectroscopic imaging. AB - Surgery is a safe and effective treatment for patients with temporal lobe epilepsy (TLE) who do not respond adequately to anticonvulsant medication and in whom the seizure generator can be identified and safely removed. Proton MR spectroscopic imaging (MRSI) can image and quantify neuronal damage in patients with TLE based on reduced signals from N-acetylaspartate (NAA), a compound localized exclusively in neurons. We performed proton MRSI in patients with TLE before and after surgical treatment to determine whether NAA or other resonance intensities changed in the temporal lobes of patients with TLE after surgery, and whether these changes correlated with surgical outcome. N-acetylaspartate resonance intensity relative to creatine (NAA/Cr) was abnormally low preoperatively in at least one temporal lobe in all 14 patients examined. It was low ipsilaterally in the patients who became seizure free and bilaterally in those who did not. Postoperatively, it increased to the normal range on the side of surgery in all patients who became seizure free. In the one patient who became seizure free and who had low NAA/Cr in both temporal lobes before surgery, NAA/Cr values in the contralateral, unoperated temporal lobe also increased to the normal range. In contrast, NAA relative intensity ratios did not change in those patients who continued to have seizures after surgery. The creatine resonance intensity (Cr) in the temporal lobes was high, relative to the brainstem, in seven patients preoperatively. After surgery, the Cr remained high in two patients, both of whom continued to have seizures. We conclude that NAA (and Cr) abnormalities in TLE do not result solely from neuronal loss and gliosis but can be reversible after postsurgical control of seizures. This implies that the NAA and Cr abnormalities in patients with TLE, at least in part, are dynamic markers of both local and remote physiologic dysfunction associated with ongoing seizures. PMID- 9409341 TI - Epileptic negative myoclonus. Subdural EEG recordings indicate a postcentral generator. AB - We report a patient with epileptic negative myoclonus (ENM) presenting as status epilepticus of the right arm. The etiology was a cortical malformation in the contralateral postcentral gyrus. Electrical stimulation of the right median nerve revealed giant surface somatosensory evoked potentials. Subdural recordings, performed to plan epilepsy surgery, demonstrated that the epileptogenic zone was in the left postcentral gyrus. Repetitive left postcentral spikes were consistently followed by an EMG silent period in the right arm with a latency of about 20 to 30 msec. The silent period in the EMG reflected the negative myoclonus and lasted 30 to 340 msec, mostly between 100 and 200 msec. Spikes in other regions of the cortex did not elicit ENM. The amplitude and duration of the spikes correlated with the EMG silent period: the more cortex involved in the spike generation, the longer the duration of the silent period. This suggests that epileptic activation of postcentral cortex leads to an inhibition of tonic activity of motor neurons. Focal ENM may be related to a hyperexcitability of the postcentral cortex. PMID- 9409342 TI - Stroke mechanisms and clinical presentation in large subcortical infarctions. AB - Large subcortical infarctions may be due to cerebral embolism and cause cortical signs more frequently than small subcortical infarctions, which usually result from small-vessel disease and are not associated with cortical findings. We evaluated 51 consecutive patients with a subcortical infarct on CT that was 1.5 cm or larger for a potential carotid or cardiac source of embolism and determined how frequently aphasia, hemineglect, or gaze paresis occurred. A carotid or cardiac embolic source was identified in 63% of the total population with a carotid source occurring in 23% and a cardiac source occurring in 49%. More than one-half of the patients with hypertension or diabetes mellitus had an embolic source, whereas all patients without these risk factors had a possible carotid or cardiac source of embolism. Aphasia or hemineglect occurred in 39% of patients and gaze paresis occurred in 41%. Large subcortical strokes frequently result in a different clinical syndrome and from a different mechanism than small subcortical strokes. PMID- 9409343 TI - Subtypes of ischemic stroke in children and young adults. AB - Specific strategies for primary and secondary stroke prevention in children and young adults can only be recommended once the causes of stroke in these age groups are well described. ICD-9 codes were used to identify children aged 1 to 18 years with acute ischemic stroke. Young adults aged > 18 to 45 years were identified from the Indiana University and Northwestern University Young Adults Stroke Registries. Validated criteria were used to subtype ischemic stroke as atherothrombotic (AT), cardioembolic (CE), small-vessel (SV), other determined cause, or unknown cause. Ninety-two children and 116 young adults were identified. Stroke subtypes in children/young adults (percentages) were as follows: AT 0/16 (p < 0.001), CE 15/14 (p = 1.0), SV 0/3 (p = 0.26), other 49/44 (p = 0.40), and unknown 36/23 (p = 0.04). Children had more prothrombotic causes (25% versus 14%, p = 0.03), and young adults had more dissections (3% versus 15%, p = 0.005). Children aged 15 to 18 years had causes of ischemic stroke more similar to those in young adults. The cause of ischemic stroke is less often identified in children than it is in young adults. Children have more prothrombotic causes of stroke, and adults have more atherothrombotic causes and dissections. Lacunar strokes are rare in both children and young adults. The age of 15 years should be used to separate childhood from young-adult ischemic stroke. PMID- 9409344 TI - Some patients with intracranial aneurysms have a reduced type III/type I collagen ratio. A case-control study. AB - A reduced production of type III collagen has been reported in previous studies to be associated with intracranial aneurysms. The purpose of this prospective case-control study was to assess the possible role of a reduced type III collagen production as a risk factor for having an intracranial aneurysm. The study group consisted of 41 consecutively admitted patients with intracranial aneurysms. Intracranial aneurysms were demonstrated by intraarterial digital subtraction cerebral angiography or during operation. The control group consisted of 41 healthy volunteers matched for age and sex. Fibroblasts were cultured from skin biopsies from patients and control subjects, and the type III/type I collagen ratios were determined. The type III/type I collagen ratios in the controls ranged from 5.5 to 19.8%, with a median ratio of 10%, and none had a ratio below 5.5%. The type III/type I collagen ratios in patients ranged from 1.1 to 25.1%, with a median ratio of 10.5%, and eight patients (19.5%) had a low (< 5.5%) ratio (p = 0.005, Fisher's exact test). Our findings support the hypothesis that a reduced production of type III collagen may contribute to the formation of intracranial aneurysms in some patients. PMID- 9409345 TI - Von Willebrand factor and risk of ischemic stroke. AB - We carried out a case-control study to determine whether von Willebrand factor (vWF) antigen (and factor VII and tissue plasminogen activator [tPA] antigens) are associated with ischemic stroke. Ninety-five patients with transient ischemic attack or minor ischemic stroke recruited from the Oxfordshire Community Stroke Project and one neurology clinic were compared with 236 controls, group-matched for age and sex, from the same general practitioners as the incident cases. In crude analyses, concentrations of vWF antigen were significantly higher in cases than in controls (p = 0.004). The age- and sex-adjusted odds ratios from lowest (referent) to highest quartile of vWF antigen were 1.00, 1.15, 2.34, and 2.36 (trend test, p = 0.006). Factor VII antigen and tPA antigen were not significantly different between cases and controls. Although adjustment for other potential risk factors abolished the statistical significance of the association between vWF and disease, this was largely due to the influence of history of ischemic heart disease. We conclude that vWF is a potent and independent risk factor for transient ischemic attack, minor ischemic stroke, and, by extrapolation, ischemic stroke in general. The data also suggest that vWF may be a risk factor for both ischemic stroke and ischemic heart disease. We found no evidence to implicate factor VII and tPA as risk factors for ischemic stroke. PMID- 9409346 TI - Patterns of sensory dysfunction in lateral medullary infarction. Clinical-MRI correlation. AB - Sensory dysfunction in lateral medullary infarction (LMI) has been insufficiently studied. We prospectively analyzed the sensory signs of 50 consecutive patients with LMI, correlating them with MRI results. The classical ipsilateral trigeminal contralateral body/limb pattern was observed only in 13 patients (26%) with lesions confined to the most posterolateral part of the caudal-middle medulla, whereas the bilateral trigeminal pattern observed in 12 patients was associated with large, ventrally extending lesions usually at the middle-rostral medulla. The contralateral trigeminal pattern was observed in nine patients with lesions sparing the most posterolateral area of the medulla. Isolated body/limb and isolated trigeminal involvement were observed in 10 and four patients respectively, usually associated with very small lesions. No sensory sign was noted in two patients. In addition to impaired sensation of spinothalamic modalities, six patients had decreased vibratory sensation in the hypoalgesic body/limb, whereas four patients had a lemniscal sensory deficit on the side contralateral to the hypoalgesic body/limb. Fifteen patients showed sensory gradient or level at the body/limb, and five had delayed appearance of sensory deficits. Trigeminal sensation was usually inhomogeneously involved among three divisions, which was more often of an onion-skin pattern than a divisional pattern. The perioral area, or V3, was generally spared or less severely involved on the side contralateral to the lesion. The sensory manifestations of LMI are extremely diverse and usually, although not always, correlate with MRI findings. The so-called classic, dissociated sensory pattern is actually uncommon, whereas sensory patterns previously thought of as atypical are relatively frequent. PMID- 9409347 TI - Pallidal stimulation for Parkinson's disease. Two targets? AB - There has been renewed interest in functional surgery as treatment for Parkinson's disease (PD). Although pallidotomy and chronic pallidal stimulation are highly effective in suppressing levodopa-induced dyskinesia (LID), both methods also seem to be effective in reducing parkinsonian disability. However, the simultaneous improvement of LID and motor signs is hard to explain with the classic model of basal ganglia circuitry. Taking advantage of the fact that deep brain stimulation is reversible and that implanted electrodes contain four discrete stimulation sites, we investigated the effect of stimulation on different sites of the globus pallidus (GP) in five PD patients. Stimulation in the dorsal GP (upper contact) significantly improved gait, akinesia, and rigidity and could induce dyskinesia when patients were in the "off" state. In contrast, stimulation in the posteroventral GP (lower contact) significantly worsened gait and akinesia, although the reduction in rigidity remained. For patients in the "on" state, stimulation in the posteroventral GP dramatically reduced LID but, as in the "off" state, worsened gait and akinesia, thus canceling out the antiparkinsonian effect of levodopa. Our results indicate that stimulation had a striking different effect on parkinsonism and dyskinesia when applied at two different loci of the GP and that stimulation applied in the posteroventral GP produced opposite effects on rigidity and on akinesia. We conclude that parkinsonian signs and LID are a reflection of at least two different anatomofunctional systems within the GP and that this functional organization of the GP needs to be considered when determining the optimal target for surgical treatment of PD. PMID- 9409348 TI - The relationship between dementia and direct involvement of the hippocampus and amygdala in Parkinson's disease. AB - Severe dementia affects 10 to 20% of all patients with Parkinson's disease (PD) and is particularly common in those aged 65 years and over. In a clinicopathologic study, we correlated Mini-Mental State Examination scores and DSM-III dementia ratings with the density of Lewy bodies, Lewy neurites, neurofibrillary tangles, neuritic plaques, gliosis, and neurons in the hippocampus and amygdala of 27 PD patients without Alzheimer's disease changes. Cortical Lewy body densities were examined in the anterior cingulate gyrus. The degree of cognitive impairment was correlated with the density of Lewy neurites in the CA2 hippocampal field, raising the possibility that disruption of the connection between the dentate gyrus, entorhinal cortex, septal nuclei, and hypothalamus and the CA1 field contributes to dementia in PD. PMID- 9409349 TI - The dopamine transporter gene and Parkinson's disease in a Chinese population. AB - We studied a variable number tandem repeat polymorphism within the dopamine transporter gene (DAT) for an association with Parkinson's disease in a Chinese population. Five alleles were detected, consisting of 6, 8, 9, 10, and 11 copies of the 40 base pair repeat sequence. The 10-copy allele was most common, accounting for 90% of alleles. There were no significant differences between the patients and the control subjects in the distribution frequencies of the alleles or genotypes. Therefore, this polymorphism is not associated with Parkinson's disease in Chinese populations. PMID- 9409350 TI - Metric properties of nurses' ratings of parkinsonian signs with a modified Unified Parkinson's Disease Rating Scale. AB - We evaluated the ability of nurse clinicians to assess parkinsonian signs in older persons with a modified version of the motor section of the Unified Parkinson's Disease Rating Scale (UPDRS). After completing a structured training protocol, three nurse clinicians and a neurologist with expertise in movement disorders administered a modified UPDRS to 75 older persons. The nurses repeated the assessment about 3 weeks later. Inter-rater agreement and short-term temporal stability were estimated for each item, the total modified UPDRS score, and for summary measures of bradykinesia, postural reflex impairment, rigidity, and tremor, and a global parkinsonian sign score. We performed our assessment in Catholic religious communities in the Chicago area, using consecutive subjects at four communities participating in the Religious Orders Study, a longitudinal, clinical-pathologic study of older persons. Our results showed that nurses were not a significant source of variability, with intraclass correlations exceeding 0.97 for all items, and they showed good to excellent agreement with the neurologist for most modified UPDRS items. Correlations between nurses and neurologist exceeded 0.90 for the total modified UPDRS, ranged from 0.76 to 0.95 for the four parkinsonian domain scores, and exceeded 0.90 for the global parkinsonian sign score. Nurses showed fair to good agreement with themselves over the 3-week interval for most modified UPDRS items. Correlations over the 3 week interval exceeded 0.90 for the total modified UPDRS score, ranged from 0.70 to 0.95 for the four domain scores, and exceeded 0.90 for the global parkinsonian sign score. Ratings of parkinsonian signs by nurse clinicians corresponded closely to those of a neurologist with expertise in movement disorders and showed good inter-rater agreement and temporal stability. With appropriate training, nurse clinicians can reliably administer the modified UPDRS. PMID- 9409351 TI - Clozapine in Parkinson's disease tremor. Effects of acute and chronic administration. AB - The effects of the acute administration of clozapine on parkinsonian mixed tremor (i.e., resting and postural tremors) were evaluated to establish clozapine's predictive value for long-term response and to determine if there is a difference in the pharmacologic responses of the two tremors. We also investigated the correlation between reduction of tremor and induction of sedation after acute and chronic administration of clozapine. Clozapine (12.5 mg) or placebo were administered po in a double-blind manner to 17 PD patients with mixed L-dopa resistant tremors. Two patients did not reach 50% improvement and were considered nonresponders. The remaining 15 patients reported moderate to marked reduction of tremor. Responsive patients in the acute test moved on to a long-term, open clozapine add-on study receiving an average daily dose +/- SD of 45 +/- 9.6 mg for a period of 15.5 +/- 8.3 months. A significant reduction of both resting (p < 0.05) and postural (p < 0.05) tremors was observed under clozapine from the first week of treatment through the entire period of the study. There was no statistically significantly difference between the degree of improvement for resting and postural tremors after either single or chronic clozapine administration. Sedation was the only side effect reported after clozapine; however, the time courses of sedation and tremor reduction did not coincide in the acute or in the chronic experimental paradigm, where it decreased considerably in a few weeks in all patients. During long-term clozapine treatment, neither systemic side effects nor worsening of motor disability scores were noted. Thus we wish to propose an acute test or a therapeutic attempt, or both, with clozapine before defining a case of mixed parkinsonian tremor as resistant tremor and therefore resorting to a neurosurgical approach. PMID- 9409353 TI - Multiple-system atrophy is genetically distinct from identified inherited causes of spinocerebellar degeneration. AB - Multiple system atrophy (MSA) is a neurodegenerative disorder of unknown cause. The only case-control study conducted in MSA patients to date suggested a possible contributory genetic component in the pathogenesis of this disorder. The aim of this study was to evaluate a possible overlap between clinically or pathologically well-defined MSA and other conditions with an identified genetic defect causing spinocerebellar degeneration in humans or mutant mice strains. The spinocerebellar ataxia type 1 and 3 genes (SCA1 and SCA3) were analyzed for a pathologic expansion in 80 patients with MSA to evaluate a possible overlap between MSA and SCA1 or SCA3. Weaver mice and lurcher mice are animal models for spinocerebellar degeneration; both share pathologic features with MSA. We sequenced the H5 pore region of the human homologue of the weaver mouse gene, hiGIRK2, in all our patients. In lurcher mice, previous biochemical studies have shown a decreased intracellular response to insulin-like growth factor 1 (IGF-1) in the cerebellar cortex, and we thus investigated the possibility of an allelic association between MSA and the receptor for IGF-1. In addition, we evaluated a possible involvement of the ciliary neurotrophic factor gene (CNTF) and examined the role of HLA-A32 to clarify the conflicting data from previous studies. No changes were detected in any of the analyzed genes. Our studies strongly suggest that MSA is an autonomous syndrome distinct from identified genetic causes for spinocerebellar degeneration. PMID- 9409352 TI - Intrathecal baclofen therapy for stiff-man syndrome and progressive encephalomyelopathy with rigidity and myoclonus. AB - We report on eight patients with stiff-man syndrome (SMS) or its "plus" variant, progressive encephalomyelopathy with rigidity and myoclonus (PERM) receiving intrathecal baclofen via pump. In six of the patients, follow-ups continued for approximately 2.5 to 6.5 years after pump implantation. Intrathecal baclofen was an effective last-resort alternative for patients who responded poorly to or did not tolerate oral antispasticity medications. General mobility increased, and spasms and rigidity were reduced; however, no complete remissions were observed either before or after pump implantation. PERM patients showed more severe and rapid progression of symptoms and more attacks of autonomic dysregulation than SMS patients. They also required higher doses and more rapid dosage increases. Complications of intrathecal baclofen therapy included spasm-induced rupture of the catheter, catheter dislocation causing radicular symptoms, and pump malfunction resulting in inaccurate dosage administration. Patients suffered fewer side effects with intrathecal baclofen than with oral medication, but overdose resulted in a transient, comalike state in one patient and sudden dosage reduction due to pump failure was fatal in another. PMID- 9409354 TI - Atrophy of the cerebellum and brainstem in dentatorubral pallidoluysian atrophy. Influence of CAG repeat size on MRI findings. AB - To elucidate how the size of the expanded CAG repeat of the gene for dentatorubral pallidoluysian atrophy (DRPLA) and other factors affect the atrophy of the brainstem and cerebellum, and the appearance of high-intensity signals on T2-weighted MRI of the cerebral white matter of patients with DRPLA, we quantitatively analyzed the MRI findings of 26 patients with DRPLA, the diagnosis of which was confirmed by molecular analysis of the DRPLA gene. When we classified the patients into two groups based on the size of the expanded CAG repeat of the DRPLA gene (group 1, number of CAG repeat units > or = 66; group 2, number of CAG repeat units < or = 65), we found strong inverse correlations between the age at MRI and the areas of midsagittal structures of the cerebellum and brainstem in group 1 but not in group 2. Multiple regression analysis, however, revealed that both the patient's age at MRI and the size of the expanded CAG repeat correlated with the areas of midsagittal structures. Involvement of the cerebral white matter as detected on T2-weighted images was observed more frequently in patients belonging to group 2 than in group 1 patients. Furthermore it was demonstrated that high-intensity signals can be detected on T2-weighted images of the cerebral white matter of patients with a largely expanded CAG repeat (group 1) in their thirties. These results suggest that patient age as well as the size of the expanded CAG repeat are related to the degree of atrophy of the brainstem and cerebellum, and the white matter changes in patients with DRPLA. PMID- 9409355 TI - Familial amyotrophic lateral sclerosis. Molecular pathology of a patient with a SOD1 mutation. AB - We report the clinical, genetic, and neuropathologic findings in a patient with rapidly progressive familial amyotrophic lateral sclerosis (ALS). We detected a point mutation at codon 48 of the Cu/Zn superoxide dismutase gene (SOD1) leading to a substitution of histidine by glutamine in the copper-binding domain. The histopathologic features are consistent with those described in rapidly progressive sporadic ALS and do not support claims that sporadic and familial disease are different pathologic entities. Neurofilamentous accumulations, hyaline, and ubiquitinated inclusions were present in the motor cortex, brainstem, and anterior horn cells, but there was no evidence of abnormal SOD1 immunoreactivity. This confirms that the cytoskeletal pathology specific to ALS is secondary to an unknown biochemical disturbance caused by mutant SOD1 molecules and not its toxic accumulation. PMID- 9409356 TI - Broadened Friedreich's ataxia phenotype after gene cloning. Minimal GAA expansion causes late-onset spastic ataxia. AB - We describe three siblings from an Italian family affected by an autosomal recessive spinocerebellar degeneration. Gait ataxia, presenting between 38 and 45 years, was the first symptom in all three patients. Dysarthria, dysmetria, brisk tendon reflexes, extensor plantar response, and scoliosis were constant features. Disease progression was slow. Electrophysiologic studies demonstrated a slight reduction in sural nerve sensory action potential in only one patient. Analysis of GAA expansion within the X25 gene showed that patients were homozygous for the expansion, with the shorter expanded allele ranging from 120 to 156 triplets. The size of the GAA expansion may be smaller than we previously described. Such minimal expansions may result in atypical forms of Friedreich's ataxia. PMID- 9409358 TI - Linkage mapping of the gene for Charcot-Marie-Tooth disease type 2 to chromosome 1p (CMT2A) and the clinical features of CMT2A. AB - Charcot-Marie-Tooth disease type 2 (CMT2) is characterized by a motor conduction velocity of the median nerve of > 38 m/sec and is a genetically heterogeneous disorder with at least three loci identified: CMT2A (1p35-36), CMT2B (3q13-22), CMT2C (not linked to any known loci), and CMT2D (7p14). In this study, we performed linkage analysis of two Japanese CMT2 families using markers flanking the CMT2A, CMT2B, and CMT2D loci. The highest cumulative multipoint lod score of 3.69 was obtained at D1S244. The CMT2B and CMT2D loci were excluded by the results of linkage analysis performed using markers D3S1551, D3S1290, and D7S484. The clinical features of the CMT2A affecting the two families include similar levels of muscle weakness of the posterior and anterior tibial muscles, tendon reflexes preserved in upper extremities but reduced or absent in lower extremities, no enlargement of the peripheral nerves, and mild sensory disturbance in only 20% of affected individuals. PMID- 9409357 TI - Effect of recombinant human insulin-like growth factor-I on progression of ALS. A placebo-controlled study. The North America ALS/IGF-I Study Group. AB - The objective of this study was to investigate the safety and efficacy of recombinant human insulinlike growth factor-I (rhIGF-I) in the treatment of sporadic ALS. A double-blind, placebo-controlled, randomized study of 266 patients was conducted at eight centers in North America. Placebo or rhIGF-I (0.05 mg/kg/day or 0.10 mg/kg/day) was administered for 9 months. The primary outcome measure was disease symptom progression, assessed by the rate of change (per patient slope) in the Appel ALS rating scale total score. The Sickness Impact Profile (SIP), a patient-perceived, health-related quality of life assessment, was a secondary outcome variable. Progression of functional impairment in patients receiving high-dose (0.10 mg/kg/day) rhIGF-I was 26% slower than in patients receiving placebo (p = 0.01). The high-dose treatment group was less likely to terminate the study due to protocol-defined markers of disease symptom progression, and members in this group exhibited a slower decline in quality of life, as assessed by the SIP. Patients receiving 0.05 mg/kg/day of rhIGF-I exhibited trends similar to those associated with high-dose treatment, suggesting a dose-dependent response. The incidence of clinically significant adverse experiences was comparable among the three treatment groups. Recombinant human insulin-like growth factor-I slowed the progression of functional impairment and the decline in health-related quality of life in patients with ALS with no medically important adverse effects. PMID- 9409359 TI - Gene dosage effects in hereditary peripheral neuropathy. Expression of peripheral myelin protein 22 in Charcot-Marie-Tooth disease type 1A and hereditary neuropathy with liability to pressure palsies nerve biopsies. AB - A duplication of a 1.5-Megabase genomic region encompassing the gene for the peripheral myelin protein 22 (PMP22) is found on chromosome 17p11.2-12 in Charcot Marie-Tooth disease type 1A (CMT1A), whereas the reciprocal deletion is associated with hereditary neuropathy with liability to pressure palsies (HNPP). Since most CMT1A patients harbor three copies of the PMP22 gene, and most HNPP patients carry only a single copy, a gene dosage effect has been proposed as a mechanism for both diseases. We have analyzed the steady-state expression of PMP22 protein in sural nerve biopsies from three CMT1A and four HNPP patients. Quantitative immunohistochemical determination showed that PMP22 protein expression relative to that of myelin protein zero and myelin basic protein was increased in all CMT1A patients and reduced in all HNPP patients, as compared with biopsy samples of patients with normal PMP22 gene expression. These data demonstrate that both neuropathies result from an imbalance of PMP22 protein expression. PMID- 9409360 TI - A case-control and nerve biopsy study of CREST multiple mononeuropathy. AB - From 536 patients with the CREST syndrome (calcinosis, Raynaud's phenomenon, esophageal dysmotility, sclerodactyly, and telangiectasis), seven were identified as having peripheral neuropathy not attributable to another cause. Peripheral neuropathy developed 0 to 25 years after their first symptoms of scleroderma. Unexplained neuropathy in CREST patients (seven patients) was more frequent than in control subjects (two patients) matched for age, sex, time of evaluation, and geographic referral region. Multiple mononeuropathy occurred significantly more frequently in the CREST group (six patients) than in the control group (0 patients). Four sural nerve biopsy specimens from the CREST patients demonstrated multifocal fiber loss and perivascular inflammation; one was diagnostic for necrotizing vasculitis and two others were highly suggestive for necrotizing vasculitis. The density of myelinated fibers in three nerves from CREST patients was significantly decreased, whereas the index of dispersion (a measure of multifocal fiber loss) was increased, and the frequency of axonal degeneration was significantly increased. Based on these clinical and pathologic findings, we conclude that in the CREST syndrome multiple mononeuropathy, although occurring infrequently, occurs more frequently than by chance and necrotizing vasculitis is the cause of this multiple mononeuropathy. PMID- 9409361 TI - Anesthetic and surgical complications in 219 cases of myotonic dystrophy. AB - The objective of this study was to assess the frequency, type, and severity of perioperative complications after a first surgery under general anesthesia in patients with myotonic dystrophy (DM) and to measure the association with suspected risk factors. Numerous cases of perioperative complications in DM patients have been reported. Hazards have been associated with the use of thiopentone, suxamethonium, neostigmine, and halothane. A retrospective study of perioperative complications was conducted for 219 DM patients who had their first surgery under general anesthesia at the Chicoutimi Hospital. The overall frequency of complications was 8.2% (18 of 219). Most complications (16 of 18) were pulmonary, including five patients with acute ventilatory failure necessitating ventilatory support, four patients with atelectasis, and three patients with pneumonia. Using multivariate analysis, we found that the risk of perioperative pulmonary complications (PPC) was significantly higher after an upper abdominal surgery (odds ratio (OR), 24.4; 95% CI, 4.0 to 149.3) and for patients with a severe muscular disability, as assessed by the presence of proximal limb weakness (OR, 14.1; 95% CI, 1.5 to 134.4). The likelihood of PPC was not related to any specific anesthetic drug. Because of the increased risk of PPC, careful monitoring during the early postoperative period, protection of upper airways, chest physiotherapy, and incentive spirometry are mandatory in all symptomatic DM patients, particularly those with a severe muscular disability or those who have undergone an upper abdominal surgery. PMID- 9409362 TI - A skeletal muscle-specific form of decorin is a target antigen for a serum IgM M protein in a patient with a proximal myopathy. AB - We described a myopathy in a patient, B.J., with Waldenstrom's macroglobulinemia and a serum IgM M-protein that binds to a glycoprotein located in skeletal muscle endomysium. The objective of this study was to identify and characterize the endomysial antigen. The antigenic protein was purified using sequential differential solubility steps, size exclusion chromatography, and anion exchange chromatography. We subjected the deglycosylated protein and several proteolytic fragments to sequence analysis. We used immunohistochemistry, Western blot, and ELISA to study the binding of the IgM monoclonal and the anti-decorin core protein antibodies to the muscle antigen before and after deglycosylation. Sequence analysis revealed amino acid residues with 100% homology to human decorin. The anti-decorin core protein antibody bound to the purified antigen by ELISA and Western blot methods and bound to skeletal muscle endomysium in a histologic pattern similar to the human IgM M-protein from the patient B.J. Deglycosylation experiments revealed that the human IgM M-protein from B.J. serum recognized a carbohydrate epitope on decorin, probably containing chondroitin sulfate. A skeletal muscle-specific form of the proteoglycan decorin, with a chondroitin sulfate-like epitope, is a target antigen for the IgM M-protein in our patient with Waldenstrom's macroglobulinemia and a myopathy. These results are the first demonstration of the binding of a human IgM M-protein to an endomysial antigen. The anti-decorin IgM may be relevant to disease pathogenesis because the patient studied had a myopathy with IgM monoclonal antibodies deposited in the muscle endomysium. PMID- 9409363 TI - Outcome of pyruvate dehydrogenase deficiency treated with ketogenic diets. Studies in patients with identical mutations. AB - Inborn errors of the pyruvate dehydrogenase complex (PDC) are associated with lactic acidosis, neuroanatomic defects, developmental delay, and early death. PDC deficiency is a clinically heterogeneous disorder, with most mutations located in the coding region of the X-linked alpha subunit of the first catalytic component, pyruvate dehydrogenase (E1). Treatment of E1 deficiency hs included cofactor replacement, activation of PDC with dichloroacetate, and ketogenic diets. In this report, we describe the outcome of ketogenic diet treatment in seven boys with E1 deficiency. These patients were divided into two groups based on their mutations (R349H, three patients; and R234G, four patients, two sibling pairs). All seven patients received ketogenic diets with varying degrees of carbohydrate restriction. Clinical outcome was compared within each group and between siblings as related to the intensity and duration of dietary intervention. Subjects who either had the diet initiated earlier in life or who were placed on greater carbohydrate restriction had increased longevity and improved mental development. Based on the improved outcomes of patients with identical mutations, it appears that a nearly carbohydrate-free diet initiated shortly after birth may be useful in the treatment of E1 deficiency. PMID- 9409364 TI - Reversible cerebral hypoperfusion in Lyme encephalopathy. AB - Lyme encephalopathy (LE) presents with subtle neuropsychiatric symptoms months to years after onset of infection with Borrelia burgdorferi. Brain magnetic resonance images are usually normal. We asked whether quantitative single photon emission computed tomography (SPECT) is a useful method to diagnose LE, to measure the response to antibiotic therapy, and to determine its neuroanatomic basis. In 13 patients with objective evidence of LE, SPECT demonstrated reduced cerebral perfusion (mean perfusion defect index [PDI] = 255), particularly in frontal subcortical and cortical regions. Six months after treatment with 1 month of intravenous ceftriaxone, perfusion significantly improved in all 13 patients (mean PDI = 188). In nine patients with neuropsychiatric symptoms following Lyme disease, but without objective abnormalities (e.g., possible LE), perfusion was similar to that of the treated LE group (mean PDI = 198); six possible LE patients (67%) had already received ceftriaxone prior to our evaluation. Perfusion was significantly lower in patients with LE and possible LE than in 26 normal subjects (mean PDI = 136), but 4 normal subjects (15%) had low perfusion in the LE range. We conclude that LE patients have hypoperfusion of frontal subcortical and cortical structures that is partially reversed after ceftriaxone therapy. However, SPECT cannot be used alone to diagnose LE or determine the presence of active CNS infection. PMID- 9409366 TI - Improving interobserver variation in reporting gadolinium-enhanced MRI lesions in multiple sclerosis. AB - Gadolinium-enhanced MRI is a sensitive and objective means to monitor disease activity in multiple sclerosis (MS). We evaluated the interobserver agreement and the value of observer training in reporting enhancing lesions from serial MRI. Scans of 16 MS patients were evaluated by five inexperienced and five experienced observers before and after consensus formation and training. The number of lesions at baseline, and the number of new and persistent lesions at follow-up were scored. For each condition, weighted kappa values (kappa) and the mean average difference to the median (MADM) scores were calculated. Without training, the experienced readers showed good agreement on number of lesions at baseline and new lesions at follow-up, and moderate agreement for persistent lesions. The inexperienced readers showed poor agreement for baseline and persistent lesions, and moderate agreement for new lesions. After training, both groups reported lower absolute numbers of lesions, especially the inexperienced readers. The experienced readers showed good agreement for all lesion types, the inexperienced readers showed agreement for baseline and new lesions, and agreement was moderate for persistent lesions. In both groups MADM scores were < 0.72 for baseline and new lesions, but > 1.2 for persistent lesions. Interobserver agreement is improved by training, especially in inexperienced readers. Interobserver agreement in reporting gadolinium-enhanced lesions is high, which validates the use of serial, enhanced MRI as an outcome parameter in treatment trials in MS. PMID- 9409365 TI - Kynurenine pathway inhibition reduces neurotoxicity of HIV-1-infected macrophages. AB - The AIDS dementia complex (ADC) is a consequence of excessive immune activation driven at least in part by systemic HIV infection and probably brain infection. Quinolinic acid (QUIN) is a neurotoxic tryptophan metabolite produced by macrophages in response to stimulation with cytokines or infection with HIV-1. Consequently it has been implicated in ADC pathogenesis. However, macrophages infected with HIV-1 synthesize numerous neurotoxic substances. Therefore we conducted experiments using human fetal brain tissue to determine the relative importance of QUIN as a neurotoxin in ADC. Human macrophages were infected with HIV-1 in vitro using a viral isolate from a demented patient. 6-Chloro-D tryptophan, an inhibitor of QUIN biosynthesis, was added to half the macrophage cultures to block formation of QUIN. Supernatants containing QUIN (SQpos) or in which QUIN biosynthesis had been inhibited (SQneg) were then added to human fetal brain aggregate cultures. Toxicity was evaluated using lactate dehydrogenase efflux, trypan blue exclusion, immunohistochemistry, image analysis, and electron microscopy. Each technique showed a reduction of toxicity in SQneg-treated cultures. These studies confirm the significance of QUIN as a neurotoxin in ADC and suggest that neuroprotective strategies may have a place in the treatment of this disease. PMID- 9409367 TI - Acute stroke after coronary angiography associated with protruding mobile thoracic aortic atheromas. AB - We describe two patients who developed acute embolic stroke immediately after coronary catheterization for unstable angina. Transesophageal echocardiography (TEE) and spiral CT of the chest revealed protruding floating atheromas within the aortic arch. These cases of stroke immediately after coronary catheterization suggest that protruding floating atheromas of the thoracic aorta place patients at risk for stroke. TEE or CT might predict a risk of stroke in such patients. PMID- 9409368 TI - Paroxysmal atrial fibrillation: high frequency of embolic brain infarction in elderly autopsy patients. AB - We examined brains clinicopathologically from 54 consecutive paroxysmal atrial fibrillation (PAF) patients aged 70 years or older and compared them with those of 59 age-matched controls. Symptomatic cerebral infarctions were present in 29 PAF patients (53.7%) and in 13 controls (22.0%) (p < 0.001). Symptomatic brain infarction was 2.4 times more common in PAF cases than in PAF-free controls; approximately 60% of the infarctions in the PAF cases were judged to be cardioembolic in origin. PAF in the elderly can be an important cause for cardiogenic cerebral embolism. PMID- 9409369 TI - Dichotomizing stroke outcomes based on self-reported dependency. AB - Patients participating in a clinical trial of stroke therapy were assessed 3 months after randomization using the Barthel Index, the Modified Rankin Scale, questions on activities of daily living, and extent of recovery. Those who needed help in performing the activities of daily living and had not recovered completely were classified as dependent. Self-reported dependency had a sensitivity of 94% and specificity of 80% against the Barthel Index dichotomized at 16 or below, or a sensitivity of 85% and specificity of 87% against the Modified Rankin Scale dichotomized at 2 or above. PMID- 9409370 TI - Idiopathic granulomatous angiitis of the CNS manifesting as diffuse white matter disease. AB - A 48-year-old man presented with progressive spastic paraparesis and diffuse white matter involvement on neuroimaging that suggested a primary demyelinating disease. Brain biopsy 3 years after onset of symptoms demonstrated idiopathic granulomatous angiitis. In patients with MRI features of diffuse white matter disease of uncertain etiology, open brain biopsy of leptomeninges and parenchyma should be considered to exclude idiopathic granulomatous angiitis of the CNS. PMID- 9409371 TI - Coexistence of tics and parkinsonism: evidence for non-dopaminergic mechanisms in tic pathogenesis. AB - We report four patients with tics since childhood who presented with typical Parkinson's disease (PD). No alteration in tics occurred before or during the development of symptoms. L-Dopa/carbidopa therapy in three patients improved parkinsonism without worsening tics over 3 months to 11 years of follow-up. Our experience fails to support hypotheses exclusively emphasizing overactivity of dopaminergic function underlying tics that would predict that PD would reduce and L-dopa exacerbate tics. This emphasizes the role of other neurotransmitter systems in the pathophysiology of tics. PMID- 9409372 TI - Cerebral manifestation of Erdheim-Chester disease: clinical and radiologic findings. AB - A 33-year-old woman presented with a 3-year history of progressive numbness in the hand, cerebellar ataxia, limb weakness, nystagmus, and dysarthria. T2 weighted MRI revealed abnormal foci of increased signal intensity mimicking demyelinating plaques in the periventricular white matter, and brain 18FDG-PET scan showed increased uptake in the pons. Biopsy from a tibial lesion showed aggregates of foamy histiocytes in the intertrabecular spaces replacing the bone marrow, characteristic of Erdheim-Chester disease. The patient was treated with craniospinal radiation. After 6 months, the clinical picture was stable and the MRI was unchanged. PMID- 9409373 TI - Selective defect of baroreflex blood pressure buffering with intact cardioinhibition in a woman with familial aniridia. AB - We report a symptomatic failure of the baroreceptor blood pressure (BP) buffering mechanism in a woman with familial aniridia. Her baseline BP oscillated at 0.1 Hz, the frequency of Mayer waves, with increased amplitude on standing without orthostatic hypotension. Although sudomotor function was normal, cutaneous thermoregulatory function and BP response to Valsalva's maneuver were abnormal. The defective BP buffer mechanism suggests Mayer waves could be a sympathetic mediated cardiovascular resonance. Baroreceptor cardioinhibition was intact. We presume that the lesion is in the rostral aspect of the dorsal medulla oblongata. PMID- 9409374 TI - Anhidrosis: an unusual presentation of diabetes insipidus. AB - A 61-year-old man who had a 10-year history of anhidrosis was found to have idiopathic diabetes insipidus. He showed no spontaneous sweating or pilocarpine induced sweat response. Skin pathology showed a normal eccrine gland. Microneurography detected no skin sympathetic nerve activity. Within a month of desmopressin treatment for diabetes insipidus, sweating and skin sympathetic nerve activity returned. PMID- 9409375 TI - Guillain-Barre syndrome following allogeneic bone marrow transplantation. AB - We describe four patients who developed Guillain-Barre syndrome (GBS) following allogeneic bone marrow transplantation (BMT). All four patients had a febrile illness before developing GBS. Two patients had mild graft-versus-host disease but did not require immunosuppression. These patients add to the increasing number of BMT patients whose courses have been complicated by GBS and raise the possibility that BMT patients, especially those undergoing allogeneic BMT, may have a higher risk of developing GBS. PMID- 9409376 TI - Polysomnographic findings in a patient with the mitochondrial encephalomyopathy NARP. AB - A 23-year-old woman with the mitochondrial encephalomyopathy NARP (neurogenic muscle weakness, ataxia, and retinitis pigmentosa) presented with symptoms of obstructive sleep apnea (OSA). An overnight polysomnogram (PSG) showed apnea, EEG slowing, and a paucity of sleep spindles. The patient had a tracheostomy for OSA, and 5 months later she had normal EEG patterns and marked clinical improvement. We propose that patients with mitochondrial encephalomyopathies should have sleep evaluations if the history suggests OSA. PMID- 9409378 TI - Refractory giant cell arteritis with spinal cord infarction. AB - We report an elderly patient with aggressive steroid-refractory giant cell arteritis manifesting as myelopathy and bilateral visual loss while on treatment. Pathologically, spinal cord infarction was observed and was due to extensive necrotizing granulomatous arteritis of spinal arteries. Spinal cord damage in giant cell arteritis is rare. One prior autopsy report of spinal cord infarction in giant cell arteritis did not identify vasculitic changes in the spinal arteries. PMID- 9409377 TI - An autosomal recessive disorder with posterior column ataxia and retinitis pigmentosa. AB - We report an autosomal recessive form of ataxia that is not allelic to Friedreich's disease in six individuals from a large kindred with family origins traced to a common founder of German-Swiss descent. The disorder begins during early childhood with a concentric contraction of the visual fields and proprioceptive loss. Eventually blindness, a severe sensory ataxia, achalasia, scoliosis, and inanition develop by third decade. Inversion recovery MRIs of the spinal cord in affected individuals demonstrate a hyperintense signal in the posterior columns. Finding the gene responsible for this disorder may aid in our understanding of the mechanisms that cause sensory neuronal degeneration. PMID- 9409379 TI - A prospective clinical and electrophysiologic survey of acute flaccid paralysis in Chinese children. AB - We studied 29 children admitted to Beijing Children's Hospital (BCH) with acute flaccid paralysis between June 1991 and June 1993. Twenty-seven patients had Guillain-Barre syndrome--7 with acute inflammatory demyelinating polyneuropathy and 20 with acute motor axonal neuropathy (AMAN). Two had poliomyelitis. The most common cause of acute flaccid paralysis at BCH is the AMAN pattern of GBS. PMID- 9409380 TI - Motor fibers in the sural nerve of humans. AB - Although motor fibers in the sural nerve were already described in three individuals, this nerve is considered purely sensory. We investigated the occurrence of motor fibers in 331 sural nerves of 207 individuals. We found motor fibers in 15 nerves (4.5%) or in 13 individuals (6.2%). The identification of motor fibers in the sural nerve before nerve biopsy has important implications for the interpretation of pathologic findings. PMID- 9409381 TI - Non-AIDS-related lymphomatous meningitis: combined modality therapy. AB - Twenty-two patients (age range, 38 to 69 years; median, 60 years) with lymphomatous meningitis due to metastatic non-AIDS-related non-Hodgkin's lymphoma were treated. Cytologic responses were seen in 16 patients (73%) to first-line chemotherapy, 7 patients (58%) to second-line chemotherapy, and 2 patients (40%) to third-line chemotherapy. Median survival was 10 months (range, 3 to 24 months). PMID- 9409382 TI - Progressive myoclonus epilepsy in young adults with neuropathologic features of Alzheimer's disease. AB - Progressive myoclonus epilepsy (PME) may develop in adult life. We present two patients with PME appearing around the age of 30 years in whom the disorder represented a manifestation of Alzheimer's disease. This diagnosis must be considered in addition to possible Kufs' disease or myoclonic epilepsy with ragged red fibers (MERRF) when PME develops in young adults. PMID- 9409383 TI - American Academy of Neurology guidelines for credentialing in neuroimaging. Report from the task force on updating guidelines for credentialing in neuroimaging. PMID- 9409384 TI - American Academy of Neurology neuroimaging training guidelines. The AAN Workshop on Neuroimaging Training. PMID- 9409385 TI - Homonymous quadrantanopia respecting the horizontal meridian. A feature of striate and extrastriate cortical disease. AB - It has been proposed that homonymous quadrantanopic visual field loss that respects the horizontal meridian is a pathognomonic sign of extrastriate cortical disease. However, two patients with this sign are reported--one with a rare striate and the other with an extrastriatal lesion of the visual cortex. It is therefore hypothesized that a homonymous quadrantanopia respecting the horizontal meridian may indicate striate and/or extrastriate cortical disease. PMID- 9409386 TI - Prevalence of polyclonal gammopathy in polyneuropathy. PMID- 9409388 TI - Homonymous hemifield loss in childhood. PMID- 9409387 TI - Onset of myasthenia gravis in a patient with multiple sclerosis during interferon 1b treatment. PMID- 9409389 TI - Unilateral hypnic headache: a case study. PMID- 9409390 TI - Handedness and laterality of the viscera. PMID- 9409391 TI - Assessment of CSF levels of tau protein in patients with Alzheimer's disease and mild dementia. PMID- 9409392 TI - Delirium, not dementia, resulting from hypoparathyroidism. PMID- 9409393 TI - Clinical-neuropathologic findings in multi-infarct dementia: a report of six autopsied cases. PMID- 9409394 TI - Clinical-neuropathologic findings in multi-infarct dementia: a report of six autopsied cases. PMID- 9409395 TI - Epidemiological features of moyamoya disease in Japan: findings from a nationwide survey. AB - To estimate an annual number of patients treated for Moyamoya disease in Japan and to describe the clinico-epidemiological features, a nationwide epidemiological survey was conducted in 1995. The study consisted of two questionnaires, which were distributed to departments randomly selected, of neurosurgery, neurology and pediatrics in hospitals throughout Japan. The first questionnaire inquired the number of the patients treated in 1994 and the second one detailed clinico-epidemiological information of each patient reported. Following major epidemiological findings emerged from the study: (a) The total annual number of patients treated for Moyamoya disease was estimated as 3900 (95% confidence interval (CI) 3500-4400) in Japan 1994, with the prevalence and incidence rates of 3.16 and 0.35 per 100,000 population, respectively; (b) the sex ratio (females to males) of the patients was 1.8; (c) the peak of age distribution of the patients was observed in 10-14 years old and a smaller peak in their forties; (d) the age at onset was under 10 years old in 47.8% of the patients, but some had developed the disease at the age of 25-49 years; (e) family history of Moyamoya disease was found in 10.0% of the patients; and (f) about 75% of the patients had normal activity of daily life or working ability even before treatment. The present findings were quite comparable with those obtained in the previous nationwide epidemiological survey in 1990. PMID- 9409396 TI - Epidemiological survey of moyamoya disease in Korea. AB - To both clarify the current clinico-epidemiological features of Moyamoya disease in Korea as well as compare these cases with Japanese patients, 451 cases were collected from 26 Korean major neurosurgical institutes and 296 definite cases were analyzed statistically. Although the Korean age distribution patterns of Moyamoya disease showed two peaks, similar to Japanese patients, the Korean pattern was shifted to the right thus indicating Korean adult population to be 20% higher than that of Japanese patients. The female/male ratio was 1:3, which was slightly less than that for Japanese. The family occurrence rate in Koreans was 1.8%. The incidence of cerebral infarction and bleeding in Koreans was higher while transient ischemic attack (TIA) and seizure were less than that of Japanese. The incidence of infarction in children and of hemorrhage in children and adults were also statistically higher in Koreans. The incidence of hemorrhage was higher in females than in males. Both the age at onset and sex affected the disease type. Single encephalo-duro-arterio-synangiosis (EDAS) was performed on 87.6% of all surgical cases. Although the incidence of bleeding was higher in Korea, the outcomes of the patients were similar to that of the Japanese patients. Although Korean Moyamoya disease showed a relatively higher incidence of hemorrhage and adult onset, the overall clinical background was similar to that of Japanese patients. PMID- 9409397 TI - Natural history of moyamoya disease: comparison of activity of daily living in surgery and non surgery groups. AB - The present study was undertaken to reach a clearer understanding of the natural history of Moyamoya disease. Follow-up studies were performed in 88 patients with Moyamoya disease. They were divided into the ischemia group and the hemorrhage group. The activity of daily living (ADL) of each groups were followed up and compared between those who were surgically treated and conservatively managed. Ischemic manifestations were more common in the younger children and tended to be recurrent, whereas hemorrhagic manifestations were more common in the adults. Follow-up duration of the 36 patients, who were surgically treated, ranged from 6 to 86.4 months (mean: 28.8 months). During the follow-up period, ADL was improved in 17 of 31 ischemic Moyamoya patients (55%); the condition was unchanged in nine (29%); and aggravated in five (16%). Follow-up duration of the 52 patients who were managed without surgery ranged from 12 to 216 months (mean: 67.2 months). In 35 patients of the ischemia group, ADL was aggravated in 49% and improved in only 26% during the follow-up period. However, ADL was aggravated in 12% of 17 hemorrhagic patients, but improved in 53%. Our result suggest that indirect revascularization procedures are effective for prevention of recurrent ischemic attacks which is common in pediatric patients. However, the effectiveness of indirect revascularization for hemorrhagic Moyamoya disease is not clear and requires extended follow-up study. PMID- 9409398 TI - Comparison of moyamoya disease in Japan and moyamoya disease (or syndrome) in the People's Republic of China. AB - To find out the causes of many epidemiological differences in Moyamoya disease in Japan and in the People's Republic of China, we performed a bibliographical investigation and an actual survey and reached the following conclusions: (1) Leptospiral infection can cause Moyamoya syndrome which cannot be differentiated from Moyamoya disease unless confirmed through the patients past history and a positive serum test; and (2) Moyamoya disease reported in China may include Moyamoya syndrome caused by leptospiral or other cerebral arteritis. PMID- 9409399 TI - Epidemiological study of moyamoya disease in Taiwan. AB - From January 1978 to December 1995, 92 cases of Moyamoya disease were collected from seven major medical centers in Taiwan. The data gave an annual incidence rate of 0.048 per 100,000 population. There were 40 males and 52 females and the ages ranged from 2 to 62 years with the peak incidence in the 31-40 year age group (23 cases). Cerebral infarction occurred in 20 out of 24 juvenile patients (83%), and in 24 out of 68 adult patients (35%). The difference was statistically significant. Haemorrhagic stroke was more frequent in adult patients. Computed tomographic scans following stroke showed cerebral infarction in 44 cases, ventricular haemorrhage in 26 cases, intracerebral haemorrhage in 14 cases and pure subarachnoid haemorrhage in eight. The most frequent initial symptom was motor disturbance (59%), followed by headache (49%) and impaired consciousness (35%). This survey showed an incidence rate much lower than that in Japan, but comparable with those in other Oriental countries and higher than those in Western countries. The male-to-female ratio once differed considerably from that of the Japanese series, but from the present study is now quite similar. PMID- 9409400 TI - Moyamoya disease in the United States. AB - The epidemiology and radiological features of Moyamoya disease (MMD) in the US were investigated. This study encompassed 98 cases; 26 were newly collected from eight US institutions and 72 were previously reported in the US literature. The patients ranged in age from 6 months to 67 years with age peaks in the first, third and fourth decades. MMD was seen in various ethnic groups and females were more commonly involved (71%) than males. A specific etiology could not be determined in most cases but arteriosclerosis and use of oral contraceptives were occasional associations. On angiography and/or magnetic resonance angiography (MRA), carotid arterial stenosis or occlusion was seen bilaterally in 95 cases (97%) and unilaterally in three. On MR or MRA, internal carotid steno-occlusive lesions were well demonstrated in all cases but Moyamoya collateral vessels (MMVs) were visualized in only 65% of the patients. MMVs in the basal ganglia and thalami were best demonstrated on T1 weighted images. Parenchymal lesions were seen in all patients and were often bilateral. With advances in MR techniques and increasing awareness of diagnostic guidelines, MMD will be diagnosed more frequently than before in the US. PMID- 9409401 TI - A survey of moyamoya disease in Hawaii. AB - Moyamoya disease (MMD) is a rare entity in the US with a few retrospective series and sporadic cases reported in the US and North America. Although it is known that MMD exists in all races, there is a predilection for people of Asian origin. Because of the relatively high percentage of Asians living in Hawaii, it was hypothesized that the estimated prevalence of MMD would be higher in Hawaii than the remaining US. All practicing neurologists, neurosurgeons, neuroradiologists and major hospitals in Hawaii were surveyed for MMD patients treated during the past 10 years. Medical records and angiograms (when available) were reviewed and the diagnostic guidelines for MMD from the Ministry of Health and Welfare of Japan applied. There were 53 records reviewed in 42 patients; 21 fulfilled the criteria for definite Moyamoya disease which were the focus of this study. In Hawaii, the prevalence of MMD was higher in patients of Japanese descent compared to Caucasians (P = 0.012) and higher than in the remaining US (P < 0.001). Non Japanese Asians and Pacific Islanders had a higher incidence of MMD than Caucasians that was not statistically significant. There was no difference in MMD among Japanese living in Hawaii or Japan. Males had an equal percentage of hemorrhage and infarcts; females tended to have a higher incidence of ischemic events rather than hemorrhage. Age and sex distribution of our series were similar to larger reported MMD studies. Our results suggest that: (1) Moyamoya disease in Hawaii has a higher incidence and prevalence than the rest of the US, largely due to the larger percentage of Asians, particularly Japanese, living in Hawaii; and (2) genetic rather than environmental factors may explain the increased MMD in Hawaii. PMID- 9409402 TI - Midwest experience with moyamoya disease. AB - A review of the cases of Moyamoya disease at two large Mid-Western United States Universities was undertaken for the purpose of assessing the epidemiology of Moyamoya disease. A total of 51 cases of Moyamoya disease were identified, with 12 cases classified as akin Moyamoya disease, nine cases of probable Moyamoya disease, and 30 cases of classic or definite Moyamoya disease. The conditions associated with akin Moyamoya were sickle cell disease, Down's syndrome, trauma, radiation, and neurofibromatosis. The mean age of presentation for probable and classic Moyamoya disease was 22 years. The sex predilection was approximately equal, with a slight female predominance. The racial background was identified in 22 of the definite cases, and included six patients with oriental inheritance, three with American Indian inheritance, one black, and the remainder Caucasian. Of some interest, there were five Caucasian patients with names identifiable as Eastern European in origin. The mean age of presentation of the definite Moyamoya disease was 14 years, the probable Moyamoya disease was 4 years, and the akin Moyamoya disease was 5 years. PMID- 9409403 TI - Moyamoya disease and syndrome. AB - Moyamoya disease was first described by Suzuki in 1963. The surgical management of Moyamoya disease began in the mid 1970s. At our institution we began operating on Moyamoya disease in 1979 and between 1979 and 1995 we have treated 30 children with Moyamoya disease and Moyamoya syndrome. Of these 21 children have an excellent outcome. Two children continued to have symptoms and both responded to an encephalo-myo-synangiosis (EMS) procedure, five children had a good outcome but are left with a significant neurologic deficit, one child remains in a poor state and one child had an excellent outcome but then died of his Fanconi's anemia 7 years after his encephalo-duro-arterio-synangiosis (EDAS) procedure. PMID- 9409404 TI - Risk factors of moyamoya disease in Canada and the USA. AB - Over the past 28 years, 39 patients with Moyamoya disease or syndrome defined as spontaneous occlusion of the circle of Willis with extensive basal collateral vessels have been treated by the author in Canada and the USA. All patients presented with clinical or radiologic evidence of hemorrhage (23) or ischemia and infarction (16). A total of 12 patients had associated cerebral aneurysms and seven of these patients with aneurysms presented with subarachnoid hemorrhage. The patients ages ranged from 5 to 47 years. Of these 58% were female. The patients racial origin included North American Indian, Innuit, East Indian/Pakistani, Japanese, Chinese, Filipino, Korean, Malayasian, Hispanic, African American and Caucasian. Familial clustering was seen in North American Indian, Innuit and Caucasian patients. Associated disorders (tuberculosis, pharyngitis, thalassemia, fibromuscular hyperplasia, polycystic kidney, sickle cell trait and hypertension) were common in these patients, as was the use of tobacco, alcohol and in the adult females, oral contraceptives. It may be concluded from this series that the etiology of Moyamoya disease or syndrome is probably multifactorial, but that some racial and familial groups are more susceptible. Furthermore, in that the clinical and angiographic features are identical, the separation between Moyamoya disease and syndrome may not be helpful in understanding the etiology and pathophysiology of this disorder. PMID- 9409405 TI - The main epidemiological, clinical and morphological features of moyamoya disease in Yugoslavia. AB - Over a period of 22 years, 31 Moyamoya cases have been recorded in Yugoslavia. In this group of the patients comprising four children and 27 adults, peak age incidence is in the third and fourth decades of life. Males and females have been almost equally affected. Familial cases or territorial clustering of the patients have not been noted. In most cases leading symptoms on admission were motor disturbances. In four patients unilateral involvement was observed while in 27 patients bilateral changes of the internal carotid artery were seen. PMID- 9409406 TI - Clinical and neuroradiological findings of moyamoya disease in Italy. AB - A total of 34 Italian patients (15 males and 19 females) suffering from Moyamoya disease (MMD) and selected by a questionnaire survey in 12 neuropediatric and neuroradiologic departments were studied in a multicentric study. The onset of the disease appeared either in childhood (27 patients, aged 0-16 years, mean 5.4 years) or in adulthood (seven patients, aged 25-55 years, mean 35 years). The early clinical symptoms consisted of transient ischemic attacks and/or stroke (20 cases), recurrent migraine-like headaches (seven cases), seizures (six cases) and hemorrhage (one case). A total of four familial cases were found. The final diagnosis was based in all cases on the conventional angiographic findings and more recently also on the magnetic resonance angiography (ten patients). The mean lag time between the first clinical manifestation and the angiographic diagnosis was about 2 years. A medical treatment (vasodilators, antiplatelet agents, calcium channel blockers) was followed by 21 patients, while five cases underwent a surgical revascularization. The follow-up ranges from 1 to 15 years (mean 6 years): A motor (16 cases) and/or mental impairment (14 cases) was detected especially in the childhood onset MMD; only one patient died. In nine cases the long-term outcome persisted without neurological deficit. PMID- 9409407 TI - Moyamoya disease in Europe, past and present status. AB - A questionnaire was distributed in early 1996 to 160 leading European neurological, neuro-pediatric and neurosurgical centers to assess the present status of Moyamoya disease in Europe. The response rate was 43%. Information was obtained on a total of 168 patients, of whom 110 had presented before 1992, and 58 from 1993 onward. 82% of the patients were Caucasian. In all other respects, the clinical findings were similar to those observed in Japan. The present study yields an incidence of 0.3 patients per center per year, which is approximately one-tenth of the incidence in Japan. Alongside these results, the history of the recognition and treatment of this disease in Europe is briefly discussed. PMID- 9409408 TI - Angiographic study of moyamoya disease and histological study in the external carotid artery system. AB - A total of 13 cases of Moyamoya disease were verified by internal and external carotid artery angiography. The biopsy samples were taken from the superficial temporal arteries and middle meningeal arteries in 12 and six cases, respectively. Angiographical and histological studies revealed that both of the internal and external carotid system could be involved in Moyamoya disease. Our studies demonstrated that Moyamoya disease was a type of vascular disease not only confined to the cerebral vessels. We recommend angiography of external carotid system and extracranial segment of internal carotid artery while performing three vascular angiography. Whether superficial temporal artery has been involved in angiography can be used as an indication of indirect cerebrovascular bypass. PMID- 9409409 TI - Proton magnetic resonance spectroscopy in children with moyamoya disease. AB - Proton magnetic resonance spectroscopy (MRS) is an effective method to noninvasively investigate cerebral metabolism. The aim of this study was to evaluate cerebral metabolic changes in children with Moyamoya disease using proton MRS. A total of 11 patients under 15 years old were enrolled, whose main symptoms were transient ischemic attacks. In nine patients, the more and less affected hemispheres could be determined according to their symptoms and/or single-photon emission computed tomography (SPECT). A single voxel point resolved spectroscopy was performed in the frontal or parietal white matter bilaterally by using 1.5 Tesla MR system (repetition time 2000 ms and echo time 272 ms). Voxels were located within areas that were normal on T1- and T2-weighted MR imaging. Choline (Cho), creatine (Cre) and N-acetyl aspartate (NAA) were semiquantitated as a metabolite value using an external standard reference. Pre-operative Cho, Cre and NAA values were within respective normal ranges. No metabolite showed a significant difference between the more and less affected hemispheres. A total of six patients were examined before and after revascularization surgery on one side. Pre- and post-operative comparisons demonstrated that all three metabolites showed statistically significant increases following surgery. These results suggest that, although the mechanism of spectroscopic changes needs to be further investigated, proton MRS provides a valuable method to evaluate cerebral metabolism in children with Moyamoya disease. PMID- 9409410 TI - Computer simulation of cerebral blood flow in moyamoya and the results of surgical therapies. AB - Moyamoya is the disease which involves the terminal portions of the internal carotid or origins of the middle or anterior cerebral arteries. The posterior communicating arteries are also involved, but not the vertebrals or the basilar artery. The disease occurs more commonly in females than males and it has two age peaks at less than 10 and 40 years. Over the years many treatment options or procedures have been advocated for this disease either with direct bypasses or indirect revascularization procedures or both in combination. Whether one procedure is better than another is a matter of question and still to be determined. Along with it, there are various diagnostic and research work have been done for the etiology and the management of this disease. We have tried to implement a computer model of cerebral blood flow in order to assess and predict the flow in this disease process. At this time to know and predict the effectivity of certain types of offered treatment of Moyamoya disease is only to evaluate patients clinically with long term follow ups and at some interval after surgery with angiography or blood flow determinations. This study tries to focus on the use of computerized model of predicting cerebral blood flow which tries to assess the cerebral flow and decide which treatment option would be the best for a particular patient. After various computer simulations the blood flow following each treatment option is detected and the situation which offers the best treatment in a particular case is offered to the patient. To confirm the use of utility of this computer model a larger population of patients with Moyamoya disease need to be evaluated. PMID- 9409411 TI - Cerebral hemodynamics and metabolism in moyamoya disease--a positron emission tomography study. AB - We studied the cerebral blood flow (CBF), cerebrovascular response to hypercapnia, oxygen extraction fraction (OEF), metabolic rate for oxygen (CMRO2), blood volume (CBV) and transit time (TT: CBV/CBF) in Moyamoya disease using positron emission tomography (PET). The subjects consisted of 23 patients with Moyamoya disease, including eight pediatric and 15 adult patients. Among them, ten patients were examined both before and after surgery. The pediatric patients showed a marked increase in the CBV and a prolonged TT, especially in the striatum. The adult patients also showed a prolonged TT, but it was less prominent than that observed in pediatric patients, while the CBF remained at the level of the normal controls. The cerebrovascular response to hypercapnia was markedly impaired in both the pediatric and adult patients, which thus indicates a decrease in the hemodynamic reserve capacity. After surgery, the CBV, TT and cerebrovascular response to hypercapnia all improved, mainly in the temporal cortex where extracranial-intracranial (EC-IC) bypass surgery had been performed. In the striatum, the CBV also decreased, while the TT was shortened after surgery. A PET study was thus found to be useful for evaluating the cerebral hemodynamics in Moyamoya disease and monitoring the effect of surgery. PMID- 9409412 TI - Assessment of encephalo-galeo-myo-synangiosis with dural pedicle insertion in childhood moyamoya disease: characteristics of cerebral blood flow and oxygen metabolism. AB - In the surgical treatment of childhood Moyamoya disease, we have been performing a new non-anastomotic bypass surgery using encephalo-galeo-myo-synangiosis (EGMS) with dural pedicle insertion over the brain surface. To assess the results obtained using this procedure, the long-term clinical outcome and the characteristics of cerebral blood flow and oxygen metabolism using PET were evaluated. The subjects were 19 patients with childhood Moyamoya disease admitted to our institution over 8 years from 1988 to 1996. This type of surgery was performed 39 times and the patients have been followed up from 10 months to 8 years. Preoperatively, although cerebral blood flow and oxygen metabolism were varied from individual to individual, low values of regional cerebral blood flow (rCBF) and regional cerebral metabolic rate of oxygen (rCMRO2) with relatively high values of regional oxygen extraction fraction (rOEF) were found in the frontal region in most of the patients compared to normal children. Postoperatively, obvious improvement of cerebral blood flow and hemodynamics in all the areas in the operative site and clinical symptoms and signs ameliorated or disappeared during the follow-up period. In conclusion, the present method appeared to be useful to improve the hemodynamic in the brain and its long-term clinical outcome was satisfactory. PMID- 9409413 TI - Cerebral hemodynamics in patients with moyamoya disease and in patients with atherosclerotic occlusion of the major cerebral arterial trunks. AB - To determine the difference in cerebral hemodynamics between Moyamoya disease and atherosclerotic occlusion of the major cerebral arterial trunks, we measured local cerebral blood flow (CBF) and local CO2 reactivity (CO2R) by xenon-enhanced computed tomography (CT). A total of 11 adult patients with Moyamoya disease (mean age, 39.6 +/- 7.8 years) and eight patients with atherosclerotic occlusion of the major arterial trunks (mean age, 62.4 +/- 15.4 years) were studied. Regions of interest were frontal, temporal and occipital cortex, caudate, putamen and thalamus in each hemisphere. In patients with Moyamoya disease, local CBF values in the internal carotid artery territory (frontal and temporal cortex, caudate, putamen) were significantly higher than those in the occluded side of patients with atherosclerotic occlusion. Local CO2R values in the caudate and putamen were significantly higher than those in the occluded side of patients with atherosclerotic occlusion. These results suggest that the cerebral hemodynamics of Moyamoya disease differ from those of atherosclerotic occlusion of the major cerebral arterial trunks, and may be a result of the abundant collateral circulation through basal 'Moyamoya' vessels. PMID- 9409414 TI - The clinical significance of cerebral veins in moyamoya disease. AB - A total of 11 patients with Moyamoya disease who underwent digital subtraction angiography were studied with respect to the cerebral venous system. First, delayed venous filling and the divisional change of the superficial cerebral veins have been attributed to the development of collateral pathways in Moyamoya disease and may be reflective appearances for the pathophysiological conditions in Moyamoya disease. Particularly, delayed venous filling may be an important finding which predicts an ischemic attack. Secondarily, the inferior striate veins were frequently detected in proportion to the development of Moyamoya vessels. On the other hand, the superior striate veins were prominent in the decrement stage of Moyamoya vessels. These findings may give support to the pathological findings of Moyamoya vessels that there were observed various degrees of dilatation with venous-like vessels as well as dilated perforating arteries. PMID- 9409415 TI - Quantification of regional cerebral blood flow and vascular reserve in childhood moyamoya disease using [123I]IMP-ARG method. AB - Both regional cerebral blood flow (rCBF) and regional vascular reserve (rVR) in ten childhood Moyamoya disease were quantified pre- and post-operatively by autoradiographic processing using single photon emission computed tomography (SPECT) and N-isopropyl-rho-iodoamphetamine (IMP) (IMP-ARG method) to estimate hemodynamic effectiveness of surgical revascularization. Before surgery, in two patients, rCBF was reduced in the whole territories and loss of rVR in the anterior circulation was observed; +4.3% in the anterior cerebral artery (ACA), 3.0% in the middle cerebral artery (MCA) and +17.5% in posterior cerebral artery (PCA) territories. After surgery, in eight patients without transient ischemic attack (TIA) episodes, rCBF at rest was maintained around subnormal level in the whole territories, and mean rVR was up to +11.9, +17.3 and +28.3% in ACA, MCA and PCA territories, respectively. However, rVR in the anterior circulation was significantly reduced in comparison with rVR in the posterior circulation. Quantification of both resting rCBF and rVR using IMP-ARG method could provide reliable information concerning on surgical indication and its effectiveness in childhood Moyamoya disease. PMID- 9409416 TI - Transcranial Doppler ultrasonography in patients with moyamoya disease. AB - This study included 17 patients (five men and 12 women) with clinical diagnosis of Moyamoya disease from conventional angiograms. Bilateral basal arteries were measured by the transtemporal approach with a 2 MHz pulsed Doppler instrument (TC 2 64B EME). In 28 out of the 34 basal arteries (82.4%), reliable recordings were obtained. These transcranial Doppler ultrasonography (TCD) findings were classified into three patterns: (1) High-high pattern; the mean cerebral blood flow velocity (CBFV) was increased throughout the basal arteries by over 70 cm/s (seven arteries, 25.0%); (2) high-low pattern; the mean CBFV was fastest (over 70 cm/s) at the ICA or proximal MCA and the mean CBFV decreased remarkably distally (15 arteries, 53.6%); and (3) low-low pattern; the mean CBFV was less than 40 cm/s throughout the basal arteries (arteries, 15.4%). These CBFV patterns as assessed by TCD are compared with the patients age, clinical symptoms and angiographical stagings (Suzuki's criteria). The high-high pattern on TCD was predominantly seen in the younger patients and in the earlier stages of the disease. The high-low pattern was the most common pattern of CBFV as assessed by TCD in Moyamoya patients. The low-low pattern on TCD was more common in the later stages following angiographic evaluation by Suzuki's criteria. The above patterns based on TCD findings show a good correlation with the age of the patient and the clinical diagnosis at the onset. TCD appears to be very useful in the evaluation of patients with Moyamoya disease. PMID- 9409417 TI - Intraoperative monitoring of jugular bulb oxygen saturation in patients with moyamoya disease. AB - The jugular bulb oxygen saturation (SjO2) and end-tidal carbon dioxide (ETCO2) were monitored continuously during surgery in six cases of Moyamoya disease who had demonstrated multiple episodes of transient ischemic attacks (TIAs) and/or fluctuating neurological deficits preoperatively. The arterial carbon dioxide tension (PaCO2) levels were also measured repeatedly at predetermined interval. In two cases (group H), the ETCO2 was controlled at hypercapnic levels during surgery (45.5 +/- 1.5 mmHg) and the remaining four (group N) were operated on in a normocapnic state (39.0 +/- 2.0 mmHg). The group H patients demonstrated high levels of SjO2 ranging from 72 to 85%, indicative of excessive hyperemia. One of the group H patients demonstrated mild and transient motor weakness postoperatively. The group N patients demonstrated normal levels of SjO2 ranging from 66 to 78%. All the patients in both groups demonstrated fluctuations in SjO2 levels in clear positive correlation with spontaneous changes in PaCO2 levels. The present findings indicated that: (1) Global carbon dioxide reactivity of cerebral perfusion is well preserved in patients with Moyamoya disease; and (2) hypercapnia in these patients often causes excessive hyperemia. The occurrence of postoperative neurological deficits in association with such an excessive hyperemia suggests that hyperapnia during surgery is not always beneficial. Intraoperative monitoring of SjO2 is useful for maintaining cerebral perfusion within the optimum range. PMID- 9409418 TI - Anesthetic management of children with moyamoya disease. AB - A review of the surgical and postoperative records of 127 revascularization procedures performed on 82 children with Moyamoya disease was done to evaluate changes we made in anesthetic management in response to perioperative complications. From 1982 to 1996, out of 82 children who underwent revascularization surgery at our hospital, five developed perioperative complications. One developed circulatory instability during surgery; the cause seemed to be a depth of anesthesia insufficient for preventing surgical stress. To rectify this problem, an increased dose of fentanyl was used to improve the maintenance of anesthesia. Four patients developed cerebral infarction during the early postoperative period due, in part, to inadequate management of postoperative pain. We began to administer supplemental doses of meperidine to patients after they emerged from anesthesia to provide better control of postoperative pain. Our review confirmed the effectiveness of these measures. The data suggest that during the perioperative management of children with Moyamoya disease, close attention should be paid to balancing the patients' anesthetic state against surgical stress and providing adequate postoperative analgesia. PMID- 9409419 TI - Revascularization with split duro-encephalo-synangiosis in the pediatric moyamoya disease--surgical result and clinical outcome. AB - Dural arteries are potential donor arteries for cortical revascularization. In this report, a technique of indirect anastomosis using a split dura is presented. At surgery, the dura near the branches of the middle meningeal artery was split into outer and inner layers, and the split surface of the outer layer was attached to the cortical surface (split duro-encephalo-synangiosis; split DES). This procedure, combined with standard encephalo-duro-arterio-synangiosis, was applied to 25 hemispheres in 18 patients with pediatric Moyamoya disease (mean age, 6 years). Postoperative superselective angiograms demonstrated effective cortical revascularization through the dural arteries in addition to the supply from the scalp arteries. All the patients were symptom free by 1.5 years after surgery. Postoperative reversible ischemic neurological deficit and infarction were seen in three (12%) and one (4%), respectively. The follow-up period ranged from 1 to 12 years (mean, 6.5 years). Thirteen of 16 (81%) patients led normal lives and three were mildly handicapped due to mental retardation that existed preoperatively. The split DES is a useful technique to extend the area of revascularization of ischemic hemispheres in Moyamoya disease. PMID- 9409420 TI - Combined encephalo-arterio-synangiosis and encephalo-myo-synangiosis in the treatment of moyamoya disease. AB - From January 1990 to December 1995, a total of nine cases of Moyamoya disease were treated at the National Taiwan University Hospital with combined encephalo arterio-synangiosis (EAS) and encephalo-myo-synangiosis (EMS). There were five males and four females and their ages ranged from 6 months to 31 years. Of these, two cases had their first symptom as intracranial hemorrhage and the rest of the cases had ischemic manifestations. Surgical treatment with combined EAS and EMS was performed on 16 hemispheres of the nine cases. The superficial temporal artery with its anterior and posterior branches was isolated and fixed to the pial surface. Then, the muscle pedicle from the bivalved temporal muscle was used as a dural graft to cover the artery. All the cases showed good neovascularization on follow-up angiography performed at 2-3 months after surgery. These two patients with hemorrhagic symptoms were followed for 52 and 61 months, respectively. Neither of these two cases showed recurrent bleeding. For patients with ischemic symptoms, the follow-up period ranged from 8 to 73 months (mean 41.7 months). All the patients showed improvement in their clinical symptoms. PMID- 9409421 TI - Surgical techniques and the results of a fronto-temporo-parietal combined indirect bypass procedure for children with moyamoya disease: a comparison with the results of encephalo-duro-arterio-synangiosis alone. AB - We recently treated children with Moyamoya disease using a fronto-temporo parietal combined indirect bypass procedure. Three different indirect bypass procedures (frontal EMAS, EDAS, EMS) were simultaneously carried out at three different sites. We thus treated 16 sides in 12 pediatric patients with Moyamoya disease using this method. Both the collateral formation and the improvement in the clinical symptoms were evaluated postoperatively. These results were then compared with those of the patients treated by EDAS alone. The postoperative collateral formation was more extensively seen in the patients treated with the combined bypass procedure than in those treated by EDAS alone. The improvement in ischemic symptoms was also better in the patients treated by the combined indirect bypass procedure. We therefore conclude that the combined indirect bypass procedure is more effective than EDAS alone. PMID- 9409422 TI - Direct extracranial-intracranial bypass for children with moyamoya disease. AB - To improve cerebral hypoperfusion in the ischemic type of Moyamoya disease, we have applied superficial temporal artery-middle cerebral artery (STA-MCA) double anastomoses in combination with encephalo-myo-synangiosis (EMS) for 19 hemispheres of 10 children (age from 5 to 11 years at surgery). Two branches of the STA were anastomosed to the two cortical arteries which were selected in the watershed area of the cerebral hemisphere estimated as a hypoperfusion area on the preoperative angiograms. Before surgery transient ischemic attacks (TIAs) developed from every month to every 6 months in association with hyperventilation or sobbing. No perioperative completed stroke or wound complications was observed, although single TIA developed in four patients within 1 month after surgery. Postoperative angiogram demonstrated that, not only the preoperative watershed area, but also the most of the middle cerebral artery territory was oppacified via the 2 branches of the STA in all 19 hemispheres. In a mean follow up period of 4 years, no ischemic episode was induced by hyperventilation, and there was no mental or neurological deterioration. STA-MCA double anastomoses, to the cerebral watershed area, in combination with EMS are safe and effective even for younger children with Moyamoya disease. PMID- 9409423 TI - Superficial temporal artery to anterior cerebral artery direct anastomosis in patients with moyamoya disease. AB - Some patients with Moyamoya disease require revascularization not only to the territory of the middle cerebral artery (MCA) but also to that of the anterior cerebral artery (ACA). To revascularize the ACA territory, we performed superficial temporal artery (STA)-ACA direct anastomosis in five patients with Moyamoya disease. After paramedian frontal craniotomy (approximately 5 cm diameter), STA-ACA anastomosis was carried out at the convexity using a cortical branch of the ACA as a recipient. An interposed STA graft was used in four patients: all of the grafts were shorter than 4 cm. Bypass flow was satisfactory in four patients. One patient who underwent simultaneous STA-ACA and STA-MCA anastomoses had poor bypass flow probably due to spontaneous leptomeningeal collateral channels between the ACA and MCA. Our method using a cortical branch of the ACA as a recipient and a branch of the STA for the interposed graft can be performed at the convexity and much easier than in a deep operative field. A short arterial graft does not easily occlude. The diameter of a branch of the STA is small and therefore, a size discrepancy does not present a problem. Our STA ACA anastomosis method is useful and effective for revascularization of the ACA territory in any patients with Moyamoya disease. PMID- 9409424 TI - Combined direct and indirect reconstructive vascular surgery on the fronto parieto-occipital region in moyamoya disease. AB - Between January 1992 and December 1995, eight patients with Moyamoya disease, aged from 2 to 39 years, underwent encephalo-duro-arterio-myo-synangiosis (EDAMS) on the frontal region, superficial temporal artery to middle cerebral artery (STA MCA) anastomosis combined with encephalo-myo-synangiosis (EMS) on the parietal region and encephaloduro-arterio-synangiosis (EDAS) on the occipital region using the frontal and parietal branch of the STA and the occipital artery, respectively. The development of postoperative collateral formation was assessed by carotid angiography and the improvement of clinical symptoms was evaluated for over 1 year after the bypass surgery. Of the 13 sides which underwent EDAMS and STA-MCA anastomosis with EMS, 11 sides resulted in extensive revascularization on the frontoparietal region and two sides showed localized collaterals, whereas EDAS on the occipital region demonstrated extensive and localized revascularization in each four sides and no evidence of revascularization in two sides among ten sides which underwent the EDAS. The clinical improvement due to the combined reconstructive surgery was very excellent in the reduction of the incidence of transient ischemic attacks (TIA) and reversible ischemic neurologic deficits (RIND). PMID- 9409425 TI - Direct and indirect revascularization for moyamoya disease surgical techniques and peri-operative complications. AB - We have performed surgical treatment for Moyamoya disease using the superficial temporal artery to middle cerebral artery (STA-MCA) anastomosis and encephalo duro-arterio-myo-synangiosis (EDAMS). In this paper, the surgical technique of combined revascularization for Moyamoya disease as well as peri-operative complications are discussed. Craniotomy and dural opening were extensively carried out to expose the brain surface as widely as possible. Dissection of the STA, which is the most powerful resource of direct revascularization, should be carefully carried out using a surgical microscope. The temporal muscle and middle meningeal artery, which have the most potential as sources of indirect revascularization, must be preserved. STA-MCA anastomosis to the frontal branch of the middle cerebral artery is indispensable for improving cerebral circulation of the frontal lobe. A small arachnoid membrane opening and water-tight closure are also important to avoid post-operative subdural and subcutaneous fluid collection. Ischemic events disappeared immediately after surgery in most cases. However, in several cases, transient ischemic attacks recurred for several months after the surgery. Chronic subdural hematoma was seen in two cases. PMID- 9409426 TI - Long-term intelligence outcome of post-encephalo-duro-arterio-synangiosis childhood moyamoya patients. AB - Long term post-encephalo-duro-arterio-synangiosis (EDAS) intelligence quotient (IQ) evaluated by Wechsler tests stays around the normal range (mean full intelligence quotient (FIQ), verbal intelligence quotient (VIQ) and performance intelligence quotient (PIQ) at 100.0 +/- 155, 100.0 +/- 15.7 and 100.2 +/- 163, respectively) at more than 9.5 years after the operation, provided that the patient has FIQ above 70 pre-operatively. This fact encourages us to employ operative treatment for this difficult disease in which intelligence deterioration has been reported to be a common occurrence. PMID- 9409427 TI - Moyamoya disease in adults: characteristics of clinical presentation and outcome after encephalo-duro-arterio-synangiosis. AB - To determine the clinical characteristics and the effectiveness of encephalo-duro arterio-synangiosis (EDAS) in adulthood-onset Moyamoya disease (MMD), the authors retrospectively reviewed 26 patients suffering from MMD who were admitted to Seoul National University Hospital between 1987 and 1995. When they showed major symptoms, all were more than 16 years-old. The most common presenting symptom was intracranial hemorrhage (ICrH), found in 12 patients or 46% of the total; the second was infarction and transient ischemic attack, each found in seven or 27% of them. Only one patient was found to have seizures, which were associated with a cerebral infarction. The Suzuki angiographic stage 3 and less than stage 3 accounted for 73% of all 52 hemispheres. A total of 15 patients underwent single photon emission computed tomography (SPECT) preoperatively. When the derangement of cerebral perfusion was estimated with four SPECT grades (SG), 70% of their hemispheres revealed normal (SG1) or localized decreased-perfusion (SG2). The other 30% had extensive decreased-perfusion or localized perfusion defects (SG3). There was no case who had extensive perfusion defects (SG4). A total of 17 patients underwent EDAS operations (EDAS group) and nine did not undergo any operation (no-op group). The EDAS group had significantly better clinical outcomes than the no-op group after a 12-month median follow-up period (P < 0.05). The angiographic and SPECT follow-up studies comprised six and seven cases, respectively. There was also satisfactory angiographic revascularization in all follow-up cases and improvement in cerebral perfusion at SPECT follow-up in six of seven cases. It is concluded that the involvement of posterior circulation of MMD is not frequent and cerebral perfusion is preserved in adulthood-onset MMD patients. These findings may explain the reason why hemorrhages are frequent and the late onset of symptoms in adulthood-onset MMD. Surgical treatment with EDAS seems to be effective in adulthood-onset MMD in terms of clinical improvement. PMID- 9409428 TI - Long-term results of surgically treated moyamoya disease. AB - Various surgical procedures have been tried for patients with Moyamoya disease. The most effective treatment, however, is still controversial. We retrospectively evaluated the long-term results of 71 patients (26 men and 45 women) with Moyamoya disease surgically treated in our institute. They consisted of 56 pediatric patients (younger than 15 years) and 15 adult patients. Symptoms in all patients were due to cerebral ischemia. We did 123 operations on 119 hemispheres: 18 superficial temporal artery--middle cerebral artery (STA-MCA) anastomoses, six STA-MCA anastomoses with indirect bypass (IB), 41 encephalo-duro-arterio synangiosis (EDAS), 29 encephalo-duro-arterio-myo-synangiosis (EDAMS) and 29 ribbon EDAMS. Average follow-up periods for each procedure were: 7 years for STA MCA anastomosis, 6.2 years for STA-MCA anastomosis with indirect bypass, 11 years for EDAS, 5.6 years for EDAMS and 2.6 years for ribbon EDAMS, respectively. The results of each procedure were satisfactory because the preoperative transient ischemic attacks disappeared. Analysis of follow-up angiograms shows excellent filling of the ACA and MCA territory in the patients undergoing ribbon EDAMS. However, long-term follow-up study shows that about 10% of the patients had severe difficulty in social or school life because of intellectual impairment. PMID- 9409429 TI - Familial occurrence of moyamoya disease. AB - There is extensive evidence that Moyamoya disease has a tendency for multifactorial inheritance, although the pathogenesis of Moyamoya disease is not clear. The authors report five cases showing familial occurrence of Moyamoya disease and analyse its clinical characteristics. In the past 15 years, we have encountered 68 cases of Moyamoya disease. Among these, 14 cases (10 females and four males, five family pedigrees, asymptomatic 1 case) of familial occurrence were observed. In this series, mother-to-child inheritance was observed in five cases, although there were no cases showing father-to-child inheritance. Ten patients were children with an initial onset of cerebral ischemia, at a mean age of 9.7 years. One mother was asymptomatic and two mothers had a past history of cerebral ischemia. Only one patient was a 37-year-old woman with clinical onset of intracerebral hemorrhage. There were no specific clinical characteristics in familial Moyamoya disease compared with those in sporadic Moyamoya disease. PMID- 9409430 TI - 'Angiographic moyamoya' its definition, classification, and therapy. AB - Are there any differences between probable Moyamoya disease and unilateral Moyamoya disease? What kinds of differences exist between definite and probable Moyamoya disease? Furthermore, according to the diagnostic criteria of Moyamoya disease, patients with systemic disorders and angiographic features similar to those of Moyamoya disease can not be diagnosed as Moyamoya disease. How should we call these? Such patients have been reported as 'Moyamoya syndrome,' 'quasi Moyamoya disease', 'akin-Moyamoya disease'. etc. These variations of terminology including unilateral or probable Moyamoya disease have thus led to as state of confusion. In this study the previously reported cases in the literature were surveyed to clarify how these terms have been used and how we should use them correctly in the future. Since the diagnostic criteria of this disease are mainly based on angiographic findings, the term Moyamoya 'syndrome' should not be used. A unilateral involvement without any known cause should be called 'probable.' Because some systemic diseases commonly associated with Moyamoya disease might be genetically linked, it is better to avoid using such vague expressions as 'quasi', 'akin', or 'pseudo.' There might be a coexistence of two diseases. It is therefore better to simply state that the angiographic findings are similar to Moyamoya disease, or a systemic disease with 'angiographic Moyamoya' until the etiology of the Moyamoya disease is clarified. PMID- 9409431 TI - Clinical features of probable moyamoya disease in Japan. AB - To clarify the current clinical features of probable Moyamoya disease in Japan, 180 cases were analyzed based on the cases collected by the Research Committee on Spontaneous Occlusion of the Circle of Willis of the Ministry of Health and Welfare, Japan. Although the age distribution patterns of probable Moyamoya disease showed two peaks similar to that of definite cases, the pattern shifted to the right thus indicating a reversed children versus adult ratio. The female/male ratio was 1.65, which was not significantly different from that observed in definite cases. The family occurrence rate was 6.7%. The incidence of cerebral infarction and bleeding in probable cases was higher than that in definite cases. The incidence of hemorrhage was higher in females than in males. Angiographically probable cases were at earlier stages in comparison to definite cases. The rebuild-up phenomenon in electroencephalograms was less detectable than in definite cases. Surgical procedures were performed in 63.3% of all cases and approximately 81% of them underwent bypass surgery. Single encephalo-duro arterio-synangiosis, direct bypass and combined bypass procedures were performed in 35, 40 and 25% of all surgical cases, respectively. Although the incidence of bleeding was higher in the probable cases, the outcomes of the patients were similar to that of the definite cases except for mortality. Approximately 7% of the probable cases developed into definite type within an average 6.6 year follow up period. The majority of probable Moyamoya disease cases thus seems to have somewhat different clinical characteristics from the definite disease cases. PMID- 9409432 TI - Long-term follow-up study of patients with unilateral moyamoya disease. AB - Although a number of cases of unilateral Moyamoya disease have been reported, the natural history of this disease remains unclear. The clinical features of 17 patients initially diagnosed with unilateral Moyamoya disease at our hospital are reported. Age at onset was 3-45 years (mean, 13.5). Of these 12 cases had onset of symptoms in childhood and five had onset in adulthood. Seven were male and 10 were female. An ischemic attack was the initial episode in ten of the 12 pediatric cases, two of the five adult cases presented with intraventricular hemorrhage. Of the 12 pediatric patients six developed contralateral lesions between 4 and 34 months (mean, 20) after the diagnosis of a unilateral lesion. The remaining six pediatric patients and all adult patients did not develop lesions on the normal side. The mean age at onset for patients later developing contralateral lesions was 6.2 years. The pediatric cases remaining unilateral was 7.7 years. The normal hemisphere of three of the pediatric patients has remained unchanged on repeated follow-up angiograms for over 10 years. Young children tended to develop the vascular pathology bilaterally. However, there were some pediatric cases whose normal or atypical sides remained unchanged without development of bilateral lesions. PMID- 9409433 TI - The current status of the treatment for hemorrhagic type moyamoya disease based on a 1995 nationwide survey in Japan. AB - The optimal treatment regimen for hemorrhagic type Moyamoya disease has yet to be clearly established. Furthermore, it remains unclear as to whether or not bypass surgery can help prevent future intracranial hemorrhaging in Moyamoya patients. In Japan, several treatment options, such as conservative, medical, and surgical intervention, have been employed for the treatment of hemorrhagic type of Moyamoya disease. In this study, 282 hemispheres with hemorrhagic onset were analyzed based on a 1995 nationwide survey by the Research Committee on Spontaneous Occlusion of the Circle of Willis (Moyamoya Disease) of the Ministry of Health and Welfare Japan to clarify the current status of treatment for hemorrhagic type Moyamoya disease. Questionnaires were distributed to the departments of neurology, neurosurgery, and pediatrics all over Japan asking about both treatment and rebleeding. As a result, 12.5% of the affected hemispheres were treated conservatively while 32.3% were medically treated. Ventricular drainage and/or hematoma removal was performed in 15.8%, and revascularization surgery in 38.3% of all hemorrhagic sides. Among the revascularization procedures used, 45.7% of the hemispheres underwent single indirect bypass surgery, such as EDAS while 22.2% of them received a direct (superficial temporal artery--middle cerebral artery: STA-MCA) bypass surgery. A combination of direct and indirect bypass surgery or a combination of different kinds of indirect procedures comprised 32.1%. Forty-nine out of 282 hemorrhagic hemispheres demonstrated rebleeding. An intracranial hemorrhage occurred in 15 hemispheres with ischemic onset even though nine of them had undergone bypass surgery prior to hemorrhaging. Nearly 18% of the patients with hemorrhagic type disease experienced rebleeding regardless of the treatment modalities. Based on these findings, there remains no clearly superior treatment plan for hemorrhagic Moyamoya disease to prevent rebleeding at this time. However, the selection of patients, treatment modalities, and the timing of the surgery might all play an important role in controlling rebleeding. The final outcome of the patients are therefore mainly considered to correlate with the initial severity of the clinical features. PMID- 9409434 TI - Mechanism of intracranial rebleeding in moyamoya disease. AB - Intracranial hemorrhage is the major catastrophic event in the natural course of Moyamoya disease, and outcome of the patients with rebleeding is very poor. However, the mechanism underlying intracranial rebleeding is not well elucidated. We retrospectively analyzed 15 patients who bled two times or more among 46 bled patients with Moyamoya disease. The results indicated that there were two different types in the manner of rebleeding. One group consisted of seven cases, which bled two times or more at the same site than the original bleeding site. In four of these seven cases, a ruptured aneurysm was identified at the distal part of collateral vessel or on the major vessel. In the other three cases, no source of bleeding was identified. In all of these cases, rebleeding occurred within 2 months after the initial insult except for one case. Another group consisted of eight cases, which bled repeatedly but at different sites from the initial bleeding site. In any of these cases, neither aneurysms nor other vascular abnormalities were identified. In all of these cases, rebleeding occurred more than 2 months after the initial bleeding. The present result indicated that intracranial bleeding might occur as a result of rupture of a tiny aneurysm at the periphery of collateral vessels. These aneurysms may be blown out after initial bleeding. When they persist after the event, they may rupture again in a fairly short interval. In other cases, bleeding occur at different sites from the initial site. They are considered to be a result of ruptured weak Moyamoya vessels which are forced to act as collateral pathways and are under unusually increased hemodynamic stress. PMID- 9409435 TI - Direct surgery for major artery aneurysm associated with moyamoya disease. AB - Moyamoya disease is often accompanied by intracranial major artery aneurysms in the posterior circulation which acts as collateral channels in place of the stenotic internal carotid arteries. These major artery aneurysms are considered to have high risk of enlargement and rupture due to increased hemodynamic stress. Direct surgical intervention has been recommended for the treatment of these aneurysms, but the direct approach to them is often difficult due to interference by intertwined abnormal vessels. We have performed direct surgery for seven major artery aneurysms in five patients with Moyamoya disease. Of these three aneurysms located in the anterior circulation were successfully clipped via pterional or interhemispheric approach. Of four posterior circulation aneurysms (two at the junction of the basilar artery and the superior cerebellar artery and two at the P1-P2 junction of the posterior cerebral artery), one was approached via pterional route because collateral vessels in the basal cistern was judged not to be rich on angiograms. However, the operative field was interfered by abundant fragile collateral vessels and it was difficult to reach the distal portion of the basilar artery. In contrast, in the other three cases in which the subtemporal approach was employed, there weren't any problems in exposures of the aneurysms. Our experiences indicate that subtemporal approach is superior than the pterional approach to reach the distal portion of the basilar artery in patients with Moyamoya disease. PMID- 9409436 TI - The efficacy of bypass surgery for the patients with hemorrhagic moyamoya disease. AB - The efficacy of bypass surgery for the patients with hemorrhagic Moyamoya disease were studied by multi-center retrospective questionnaire study. The rebleeding rate was 28.3% (39/138 patients) in conservative treatment group and 19.1% (29/152 patients) in bypass surgery group with no significant statistical difference. It is still difficult to clarify the efficacy of bypass surgery for prevention of hemorrhagic attack. PMID- 9409437 TI - Hemorrhagic type moyamoya disease. AB - The clinical picture of hemorrhagic type Moyamoya disease was analyzed in 20 cases. Hematoma at the basal ganglia was noted in 40% of cases, intraventricular hemorrhage (IVH) in 30%, thalamic hemorrhage with ventricular rupture in 15% and subcortical hemorrhage in 5%. The location was undetermined in two cases (10%) due to bleeding in the pre-computed tomography (CT) era. The frequencies shown above were correlated well to previous reports. In magnetic resonance imaging (MRI) performed 1 year or more after IVH, the primary bleeding site was demonstrated at the lateral wall of lateral ventricle, in portion density weighted and T2 weighted images. MRI can detect the site of old bleeding points and its chronological change if the study is repeated. In a follow-up period of 6.2 years, 35% of the cases had rebleeding once or twice. The second bleeding occurred seven times and the third twice. IVH occurred five times and the most common, basal ganglia hematoma three times while thalamic hemorrhage once. As a result, the rate of good outcome was 60% after the first bleeding and 40% after rebleeding. The mortality rate was 5% after the first 25% after rebleedings. Factors related to rebleedings and their poorer outcome are sex (with women being more susceptible), massive ICH and early recurrence. Rebleeding worsened the outcome. Therefore, prevention of rebleeding is important. From a therapeutic viewpoint, although a close relation between rebleeding and untreated hypertension could not be established, blood pressure control is critical at the both acute and chronic stages. Bypass surgery for bleeding type of Moyamoya disease seems to be less promising than ischemic type, even though a definite answer may not be obtained from our small number of cases. PMID- 9409438 TI - Histopathological studies on spontaneous occlusion of the circle of Willis (cerebrovascular moyamoya disease). AB - Spontaneous occlusion of the circle of Willis (cerebrovascular Moyamoya disease; SOCW) was first described by Japanese surgeons and is thought to have a high incidence in the Japanese population. SOCW is characterized by the angiographical findings of the obstructive vascular lesions around the terminal portions of the internal carotid arteries and the formation of abnormal vascular networks visualized in the arterial phase without any definite underlying conditions. Here, we present a detailed histopathological observation of 31 autopsy cases with SOCW to discuss its etiology which is still a matter of dispute. The obstructive vascular lesion around the terminal portions of the internal carotid arteries which is thought to be the primary site affected in SOCW is due to multilayered eccentric intimal fibrous thickening suggestive of organized mural thrombi. In fact, we found thrombotic lesions frequently in the major cerebral arteries, including those of the circle of Willis, of autopsy cases with SOCW. This fact supports the idea that thrombi play an important role in establishment of the vascular lesions. Extracranial vessels are also shown to be involved and it is conceivable that the systemic etiologic factor, such as the systemic background to form thrombi, exists in patients with SOCW. PMID- 9409439 TI - Histopathological studies on spontaneous vault moyamoya and revascularized collaterals formed by encephalomyosynangiosis. AB - Two topics concerning the leptomeningeal vessels are discussed. The first portion indirectly proves that angiographic 'vault Moyamoya' consists of dilated preexisted vessels rather than newly-formed vessels, using morphometric analysis. The second portion presents a 67 year old autopsied case with Moyamoya disease who underwent encephalomyosynangiosis (EMS). Collateral vessels fomred by EMS are histopathologically identified. PMID- 9409441 TI - Evaluation of cytokines in cerebrospinal fluid from patients with moyamoya disease. AB - We investigated the levels of angiogenic growth factors in cerebrospinal fluid (CSF) from patients with Moyamoya disease and from those with atherosclerotic occlusive disease to evaluate the relationship of these factors to the pathogenesis of Moyamoya disease. CSF from Moyamoya patients contained significantly higher concentrations of basic fibroblast growth factor (b-FGF) (P < 0.05). The b-FGF level was apparently elevated in patients with well developed neovascularization after indirect revascularization surgery (P < 0.01). The other angiogenic factors were not significantly elevated compared with those of the control group. PMID- 9409440 TI - Development of intimal thickening in superficial temporal arteries in patients with moyamoya disease. AB - To investigate the possible mechanism of neointimal formation in Moyamoya disease, we histologically examined the superficial temporal arteries and also investigated cultured smooth muscle cells (SMCs) from the arteries. Intimal thickening of the scalp arteries developed significantly at an early age in Moyamoya patients compared with control subjects. The histopathological findings of the neointima in scalp arteries were almost similar to those in intracranial arteries in Moyamoya patients. SMCs cultured from Moyamoya arteries responded significantly less to serum mitogens, especially to platelet derived growth factor (PDGF), than those of control patients, the finding of which was explained by the reduced number of PDGF receptor on Moyamoya SMCs. Our findings indicate the presence of systemic factors that promote migration and proliferation of SMCs from the media to the intima in Moyamoya disease. Our results suggest that alteration in vascular cells may contribute to the development of intimal thickening in Moyamoya disease. PMID- 9409442 TI - The relationship between moyamoya disease and bacterial infection. AB - To examine the relationship between Moyamoya disease and bacterial infections, authors studied the serum of 85 cases of Moyamoya disease and the influence of Propionibacterium acnes (P. acnes) infection on intracranial arteries in rats. The serum levels of P. acnes antibody, IgM, transferrin (Tf), alpha 2 macroglobulin (alpha 2M) were significantly higher in Moyamoya disease than in normal volunteers. Moyamoya-like changes of the intracranial internal carotid arteries were histopathologically demonstrated in P. acnes infectious rats. These findings suggest that P. acnes and immunological factors might play a role in the pathogenesis of Moyamoya disease. PMID- 9409443 TI - Studies on cytomegalovirus and Epstein-Barr virus infection in moyamoya disease. AB - In this study, measurement of Epstein-Barr virus (EBV) and cytomegalovirus (CMV) viral antibody titers and analysis of both viral genomic sequences using polymerase chain reaction (PCR) were performed to clarify the correlation of viral infection and Moyamoya disease. Serum samples were obtained from 64 patients with Moyamoya disease. The ages ranged from 5 to 66 years, with a mean age of 35.1 years. There were 23 males and 41 females. The serum antibody titers to CMV and EBV were measured by means of compliment fixation test and fluorescent antibody method respectively. Those titers of the patients were compared with those of 13 patients of atherosclerotic internal carotid occlusion and 34 normal volunteers. On the other hand, CMV and EBV genomic sequence using PCR, which was utilized with specific primer pairs, were performed in 22 patients of Moyamoya disease and ten normal volunteers. The following results were obtained; The antibody titer of EBV in Moyamoya disease was significantly higher than that in controls. However, no significant difference of antibody titer against CMV was detected. In EBV DNA analysis by use of PCR, EBV DNA was proved in 15 out of 20 patients with Moyamoya disease and four out of nine normal controls. Namely, EBV DNA was seen more frequently in patients with Moyamoya disease, compared with normal controls. In inverse, CMV DNA was not seen in patients with Moyamoya disease nor normal controls. In conclusion, the antibody titer of EBV revealed high levels in Moyamoya disease and EBV DNA was also detected more frequently in patients with Moyamoya disease. These results suggested a possibility that EBV infection might be involved in the pathogenesis of Moyamoya disease. PMID- 9409444 TI - Thrombophilia found in patients with moyamoya disease. AB - Sixteen patients with Moyamoya disease and four with quasi-Moyamoya disease were investigated in order to elucidate the presence of thrombophilia. The assay system for diagnosing thrombophilia consisted of assessing both the activity and antigen levels of antithrombin III, protein C, protein S, fibrinogen and plasminogen as well as detecting lupus anticoagulants. The analysis revealed that one third (four definite cases and three quasi-cases) of the examined patients demonstrated either congenital or acquired thrombotic tendency. Protein C deficiency was found in two definite cases and in two quasi-cases among whom one quasi-case was identified to have a hereditary type I Protein C deficiency. Protein S deficiency was found in one definite case and in one quasi-case. Type II plasminogen deficiency was found in one quasi-case, and lupus anticoagulant was present in one quasi-case. Based on these findings, an evaluation of thrombophilia should thus be performed when both diagnosing and treating suspected cases of Moyamoya disease. PMID- 9409445 TI - Analysis of class II genes of human leukocyte antigen in patients with moyamoya disease. AB - Moyamoya disease is a progressive steno-occlusive disease at the terminal portion of the bilateral internal carotid arteries. An unusual vascular network is formed as a result of ischemic change of cerebrovascular system. Although some investigations suggested possible etiological factors of Moyamoya disease, the etiology is still unknown. To elucidate the genetic factors of Moyamoya disease, class II genes of human leukocyte antigen (HLA) were analyzed at the DNA level. The DNA typing of HLA was performed in the unrelated Japanese patients with definite Moyamoya disease using extracted genomic DNA from the leukocyte. The genotype was confirmed by polymerase chain reaction-sequence specific oligonucleotide probe (PCR-SSOP) technique. The class II genotypes were analyzed in 71 samples. As a result, several alleles of class II genes showed significant association with Moyamoya disease. DQB1*0502 had positive association with the disease. On the other hand DRB1*0405 and DQB1*0401 had negative association. Moyamoya disease seems to have a genetic background in its etiology because certain alleles of HLA are associated with Moyamoya disease. PMID- 9409446 TI - Guidelines for the diagnosis and treatment of spontaneous occlusion of the circle of Willis ('moyamoya' disease). Research Committee on Spontaneous Occlusion of the Circle of Willis (Moyamoya Disease) of the Ministry of Health and Welfare, Japan. PMID- 9409447 TI - Comprehensive method for the typing of HLA-A, B, and C alleles by direct sequencing of PCR products obtained from genomic DNA. AB - Molecular testing is gradually replacing standard typing techniques in the field of HLA because it allows higher resolution, which has significant functional implications. Although several techniques have been so far described for this purpose, the definitive means to determine which alleles are present in a particular sample is to identify their sequence. We describe a simplified method for typing HLA-A, B, and C alleles by direct sequencing of polymerase chain reaction (PCR) products amplified from genomic DNA that could allow large-scale handling of samples for clinical use. The template is the product of a nested PCR. A first round of PCR amplifications from genomic DNA is performed with three different sets of primers, one pair specific for each locus. The PCR products encompass exons 2 and 3, the regions of interest to determine the allele present. These fragments are a mixture of both alleles present in one locus. In a second round of PCRs using the first fragment as template, exons 2 and 3 are separately amplified and simultaneously tailed with sequences corresponding to fluorescent labeled commercial primers. The sense and antisense sequence of each exon is obtained and compared with a database of all known HLA-A, B, or C alleles. Heterozygous positions are determined and the most probable alleles assigned. This simplified procedure has the practical advantage of allowing high-resolution typing of clinical material by utilizing the same genomic DNA used for standard molecular typing of HLA class I. PMID- 9409448 TI - MHC-peptide binding: dimers of cysteine-containing nonapeptides bind with high affinity to HLA-A2.1 class I molecules. AB - Small peptides, 8-10 amino acids long, derived from degradation of cytoplasmic proteins by a proteasome-proteinase complex, are usually presented and recognized by CD8+ cytolytic T lymphocytes (CTLs) associated with major histocompatibility complex (MHC) class I molecules. Recently synthetic peptides were used for the in vitro induction of tumor-specific CTLs, offering another strategy in the study of the immune-response repertoire and providing a new tool in cancer vaccination and immunotherapy. Peptides derived from otherwise normal proteins, overexpressed in many tumors as products of the protooncogene, may represent a target for an immune response. This is the case of HER-2/neu gene (also known as ErbB-2), encoding a cysteine-rich glycoprotein transmembrane receptor with tyrosine kinase activity (gp185neu). Recent data, demonstrating that HLA-A2.1-related peptides are able to stimulate in vitro CD8+ lymphocytes, Prompted us to study the binding to HLA-A2.1 molecules of several gp185 synthetic peptides containing a cystein residue and to define the relevance of this amino acid residue in the reduced or oxidated form of the sulfhydryl group. We found that monomers and their homodimers, linked by a disulfide bridge, bind to HLA-A2.1 molecules with overlapping affinity. These results suggest that additional amino acids of the nonapeptide do not prevent the binding and the HLA refolding through chemical or sterical interactions. This might be of particular relevance for the in vivo processing of cysteine-rich proteins. Because ErbB-2 molecules, as tumor differentiation antigens in melanoma, are cysteine-rich molecules, it may be relevant to evaluate the possible role of the cystine residues interacting with the T-cell receptor. The recognition of these heterodimers by CD8+ lymphocytes will require functional in vivo studies. PMID- 9409449 TI - Construction and characterization of retroviral vectors for interleukin-2 gene therapy. AB - Several investigators have employed interleukin-2 (IL-2) gene transfer to enhance the immunogenicity of tumor cell vaccines. We describe in this report the construction and characterization of retroviral vectors for IL-2 gene therapy. Human IL-2 cDNA with a chimeric rat preproinsulin/IL-2 DNA leader sequence was subcloned into the pLXSN (long terminal repeat promoter) and pLNCX (cytomegalovirus [CMV] promoter) vectors to generate the plasmids pLXSN-iIL2 and pLNCX-iIL2, respectively. Human IL-2 cDNA with a chimeric human tissue factor/IL 2 DNA leader sequence was utilized to construct the vector pLXSN-tIL2. The levels of IL-2 secreted by transduced tumor cells and fibroblasts were evaluated by enzyme-linked immunosorbent assay (ELISA) of culture supernatants and compared with those of normal peripheral blood mononuclear cells (PBMC) activated in vitro with calcium ionophore and phorbol 12-myristate 13-acetate. The highest levels of IL-2 secreted by transduced tumor cells (760 units/10(6) cells/24 h), adult fibroblasts (625 units/10(6) cells/24 h), and embryonic fibroblasts (3,975 units/10(6) cells/24 h) were 150- to 1,000-fold higher than than secreted by the activated PBMC (4 units/10(6) cells/24 h). Similar levels of IL-2 were expressed by human fibroblasts transduced with pLXSN vectors employing the preproinsulin (pLXSN-iIL2) or tissue factor (pLXSN-tIL2) leader sequences (range in IL-2 units/10(6) cells/24 h pLXSN-iIL2 = 375-625 vs. pLXSN-tIL2 = 90-440). Because IL 2-transduced cells for clinical applications are generally irradiated to prevent cellular proliferation, we evaluated the effects of radiation on IL-2 production. Radiation doses between 1,500 and 10,000 cGy resulted in gradual decreases in IL 2 secretion by transduced cells. The range of the decrease in IL-2 secretion was 7-11% by day 7, 0-29% by day 14, and 25-50% by day 35. For clinical applications, stable production of the vector in high concentrations is an important consideration. The retroviral vector pLXSN-tIL2 produced the highest viral titer and was chosen for further characterization. Southern blot analysis of SacI digested genomic DNA from the LXSN-tIL2 producer cell line and SacI-digested pLXSN-tIL2 plasmid DNA revealed the expected 3.2-kbp fragment, suggesting the absence of transgene rearrangement and the suitability of this vector as a candidate for clinical applications. PMID- 9409450 TI - Immunization with a syngeneic tumor infected with recombinant vaccinia virus expressing granulocyte-macrophage colony-stimulating factor (GM-CSF) induces tumor regression and long-lasting systemic immunity. AB - A recombinant vaccinia virus encoding the gene for granulocyte-macrophage colony stimulating factor (rV-GM-CSF) was used to infect the poorly immunogenic murine colon adenocarcinoma cell line, MC-38. Infection of MC-38 tumor cells with rV-GM CSF completely suppressed the growth of the MC-38 primary tumors, whereas progressively growing tumors were formed in mice injected with MC-38 cells infected with wild type V-Wyeth. Irradiation of the recipient B6 mice before implantation of rV-GM-CSF-infected tumor cells resulted in the development of progressively growing tumors. Moreover, in vivo T-cell depletion studies revealed that growth suppression of the rV-GM-CSF-infected tumor cells was dependent on the presence of both CD4+ and CD8+ T-cell subsets. Subsequent studies established that this immunity was long-lasting and antigen specific, as demonstrated by the protection of rV-GM-CSF-immunized mice from MC-38 tumor challenge but not from challenge with another syngeneic tumor cell type. No such effects were observed when MC-38 tumor cells were infected with recombinant vaccinia viruses expressing interleukin (IL)-2 or IL-6. The results demonstrate that paracrine release of biologically active murine GM-CSF by tumor cells infected with rV-GM-CSF enhances the intrinsic immunogenicity of a poorly immunogenic murine tumor. Presumably the augmentation of tumor immunogenicity induces an antigen-specific T-cell-dependent antitumor response that prevents the formation of primary tumors and protects mice from tumor challenge. Thus in this experimental model, GM-CSF functions as a highly effective vaccine adjuvant. PMID- 9409451 TI - Differential expression of MART-1 in primary and metastatic melanoma lesions. AB - Twenty-eight primary and 29 metastatic melanoma lesions and 18 pigmented nevi lesions were analyzed by using the immunoperoxidase reaction with anti-MART-1 and anti-gp100 monoclonal antibodies (mAbs). The MART-1 was expressed in 28, 29, and 18, and gp100 was expressed in 27, 28, and eight of these lesions, respectively. Intensity and percentage of stained cells with anti-MART-1 mAb were stronger and higher than those with anti-gp100 mAb. MART-1 was expressed homogeneously in primary melanoma and pigmented nevi, whereas it was heterogeneously expressed in metastatic melanoma lesions. The level of expression of MART-1 in primary melanoma lesions did not correlate with any clinicopathologic parameters. These results suggest that anti-MART-1 mAb is a useful tool for immunohistochemical analysis of melanocytic lesions and also is useful for patients' selection and monitoring of antigen-loss variants in clinical trials with the MART-1-based immunotherapy. PMID- 9409452 TI - Melan-A/MART-1 antigen expression in cutaneous and ocular melanomas. AB - The human immune repertoire appears to be capable of recognizing normal antigens expressed by tumor cells. Among these antigens, those of differentiation, characterized by a restricted tissue expression, could be of clinical interest since they may represent a target for immunotherapeutic protocols. In this context we have evaluated, in benign and malignant lesions of the melanocytic lineage, the expression of the Melan-A/MART-1 antigen, which has been shown to be recognized by T cells, of HLA-A2 melanoma patients. The immunohistochemical analysis conducted with a Melan-A/MART-1 monoclonal antibody demonstrated that the antigen expression does not correlate with transformation or tumor progression. At variable levels Melan-A/MART-1, differently from other differentiation antigens, is homogeneously expressed by multiple autologous metastases and by melanoma metastases at different body sites. This tissue distribution adds further biological support to the ongoing use of Melan-A/MART-1 related peptides in active immunotherapy. PMID- 9409453 TI - Clonality of tumor-infiltrating lymphocytes in human urinary bladder carcinoma. AB - The immune system has been implicated in the control of bladder tumor growth. To evaluate the clonality of bladder tumor-infiltrating T lymphocytes (TILs) in vivo, we studied the T-cell antigen receptor (TCR) repertoire in tumor biopsy specimens from 10 patients with transitional-cell carcinoma (TCC) of the bladder. Nine patients had a primary tumor, and one had a multifocal disease, consisting of two bladder tumors and three bilateral upper urinary tract sites of involvement. The following specimens from the nine patients with a primary tumor also were analyzed: a recurrent tumor from four patients, a metastatic lymph node from one patient, and peripheral blood from five patients. We used a high resolution polymerase chain reaction (PCR) method to determine CDR3 (complementarity-determining region 3) size lengths of TCR beta-chain transcripts. Oligoclonal T-cell expansion was identified in all specimens, with a larger number of expanded clones in the tumors than in peripheral blood. Expanded clones were identified in several beta-chain variable region (BV) subfamilies and varied from one patient to the next and also in different specimens from the same patient. However, a number of clones with the same VJ combination and the same CDR3 size were identified in a given patient (in specimens collected either simultaneously or at different times), suggesting homogeneity in the immunogenic environment. Clonal T-cell expansion in patients with bladder cancer may reflect prolonged exposure of T lymphocytes to tumor antigens. Our findings provide a basis for functional studies to elucidate T lymphocyte-bladder tumor cell interactions. PMID- 9409454 TI - Defective granzyme B gene expression and lytic response in T lymphocytes infiltrating human renal cell carcinoma. AB - Granzyme B is a protein thought to play a pivotal role in the cytolytic functions of T cells. In view of this, the inducibility of this gene in freshly isolated T cells (T-TILs) infiltrating human renal cell carcinoma (RCC) in vitro was examined by using the reverse transcriptase-polymerase chain reaction (RT-PCR). A reduction in granzyme B messenger RNA (mRNA) expression in stimulated T-TILs from five of nine patients with RCC compared with autologous peripheral blood T cells was noted. The reduced expression was observed after multiple stimuli including anti-CD3 antibody, interleukin-2 (IL-2), and phytohemagglutinin (PHA). Because CD8+ T cells represent the predominant cytotoxic population, the ability of this cell population to express granzyme B mRNA after stimulation also was examined. When compared with CD8+ peripheral blood lymphocytes (T-PBLs) from patients with RCC and normal donors, the induction of granzyme B mRNA was reduced in CD8+ T TILs. CD8+ T-TILs also had lower non-major histocompatibility complex (MHC) restricted cytotoxic activity than did CD8+ T-PBLs against both Daudi cells and allogeneic RCC cell lines. These results show that in a subset of patients with RCC, depressed lytic activity of CD8+ TILs compared with CD8+ PBLs is present. Reduced granzyme B mRNA expression also was noted in selected patients. PMID- 9409455 TI - Tumor-infiltrating lymphocytes in patients with metastatic melanoma receiving chemoimmunotherapy. AB - Biopsies from 12 patients with progressive metastatic melanoma were excised during chemoimmunotherapy to evaluate and characterize the local immune response in situ in the metastases. These findings were compared with the distribution of lymphocyte subsets in the peripheral blood and correlated with the clinical data. The biopsy specimens were prepared for microscopic procedures, and the fields for analyses were chosen to involve a section of both stroma and the tumor area. The number of each lymphocyte subset was calculated and compared with the number of melanoma cells in the field, allowing quantitative characterization of the immune reaction in different samples. Comparison of the lymphocyte subsets of peripheral blood and metastatic lesions revealed equal relative amounts of CD4+ (helper) and CD8+ (suppressor/cytotoxic) cells in both tissues, but 10- to 20-fold fewer CD56+ (natural killer, NK) cells, and a total absence of CD20+ (B) cells in the metastatic lesions. The prognosis of patients was viewed at different stages of the disease. The median survival from the primary diagnosis of patients with a tumor CD4+/CD8+ ratio above the median was 4.4 years compared with 2.4 years for those with a ratio below the median (Logrank, p = 0.02). In the multivariate analysis, the only statistically significant prognostic factors were the CD4+/CD8+ ratio of the tumor (p = 0.010) and of the peripheral blood (p = 0.020). Monitoring of CD4+ and CD8+ cells may thus provide valuable information about the state of host defense, with a high CD4+/CD8+ ratio indicating more favorable prognosis. PMID- 9409456 TI - Chimeric bispecific OC/TR monoclonal antibody mediates lysis of tumor cells expressing the folate-binding protein (MOv18) and displays decreased immunogenicity in patients. AB - The bispecific OC/TR monoclonal antibody (mAb) cross-links the CD3 molecule on T cells with the human folate-binding protein (FBP), which is highly expressed on nonmucinous ovarian carcinomas. Clinical trials of patients with ovarian carcinoma with the OC/TR mAb have shown some complete and partial responses. Most patients developed human anti-murine immunoglobulin antibodies (HAMA), which can inhibit OC/TR mAb-mediated lysis. We generated a chimeric version of the OC/TR mAb to decrease the immunogenicity of the OC/TR mAb and to allow more extended treatment schedules. Sp2/0 myeloma cells were transfected with chimeric heavy- and light-chain genes encoding the anti-CD3 mAb and the MOv18 mAb, respectively, which are reactive with FBP. The resulting cell line produced 80 micrograms/ml of total immunoglobulin G (IgG), of which 11.5% was the functionally active chimeric OC/TR mAb. Chimeric OC/TR F(ab')2 fragments mediated lysis of IGROV-1 ovarian carcinoma cells by human T cells at antibody concentrations of > or = 1 pg/ml. Specific lysis was still detectable at an effector-to-target cell ratio as low as 0.4. Two patients with ovarian carcinoma treated with F(ab')2 fragments of the murine OC/TR developed distinct HAMA titers, which were mainly anti-idiotypic and only partly directed against the murine antibody constant regions. However, of the two patients that were treated with the F(ab')2 fragments of the chimeric OC/TR mAb, only one developed a low transient HAMA response just above background level. In conclusion, the generation of chimeric OC/TR may allow more extended clinical studies of bispecific mAb-mediated immunotherapy of ovarian carcinoma. PMID- 9409457 TI - Six-year clinical evaluation of HTR synthetic bone grafts in human grade II molar furcations. AB - A biocompatible microporous composite of PMMA (poly-methyl-methacrylate), PHEMA (poly-hydroxy-ethyl-methacrylate) and calcium hydroxide bone replacement graft material (Bioplant HTR Synthetic Bone) was evaluated in 16 maxillary molar and 10 mandibular molar Grade II furcations in 13 patients. Following initial preparation, full thickness flaps were raised to gain access to the furcations; mechanical hand and ultrasonic root and defect debridement and chemical tetracycline root preparation were performed; furcation and adjacent osseous defects in each patient were grafted with HTR Synthetic Bone; and the host flaps replaced or slightly coronally positioned. Weekly, then monthly deplaquing was performed until surgical re-entry at 6-12 months. Patients were then followed on approximate 3-month recalls for > or = 6 yr. Evaluation of the primary clinical outcome of furcation grade change showed that in the maxilla 5/16 furcations were clinically closed, 9/16 were Grade I, and 2/16 remained Grade II; while in the mandible 3/10 were clinically closed, 5/10 were Grade I, and 2/10 remained Grade II. Other significant clinical changes included decrease in mean horizontal furcation probing attachment level from 4.4 mm at surgery to 2.2 mm at re-entry to 2.0 mm at 6 yr, decrease in probing pocket depth from 5.4 mm at surgery to 3.0 mm at re-entry to 3.2 mm at 6 yr, and improvement in vertical clinical probing attachment level from 5.4 mm at surgery to 4.2 mm at re-entry to 4.1 mm at 6 yr (all p < 0.05 from surgery to re-entry and surgery to 6 yr, n.s. from re-entry to 6 yr via ANOVA). These favorable results with HTR polymer are similar to several reports with other graft materials and with GTR barriers, and suggest that HTR polymer may have a beneficial effect in the clinical management of Grade II molar furcations. PMID- 9409458 TI - CD44 isoform expression in periodontal tissues: cell-type specific regulation of alternative splicing. AB - CD44 functions as a receptor for various extracellular matrices and plays crucial roles in homotypic and heterotypic cell-cell interactions. Recently, the molecular structure of CD44 has been extensively analyzed and multiple isoforms produced by alternative splicing of messenger RNA have been identified. In this study, we examined the expression of CD44 isoforms on different cell types isolated from periodontal tissue. In order to examine tissue differences in CD44 isoform expression, we established in vitro cell culture of human gingival fibroblasts (HGF), human periodontal ligament cells (HPDL) and human gingival epithelial cells (HGEC). These cells all expressed CD44 protein and messenger RNA. However, immunoprecipitation and Northern blot analysis revealed that HGEC expressed larger CD44 isoforms than HGF and HPDL. Reverse transcription polymerase chain reaction with primers flanking the insertion site of alternatively spliced exons was used to study details of the heterogeneity. All cells examined expressed a major band in the absence of alternatively spliced exons and additional larger bands. In particular, HGEC contained more abundant high molecular mass species. In vitro stimulation by IL-1 beta, TNF alpha or phorbol 12-myristate 13-acetate induced an increase in total CD44 messenger RNA in HGF but not change in overall patterns of CD44 isoform expression. However, the isoform expression of HGEC was sensitive to cell density. The amount of larger isoform was decreased by culturing cells beyond confluence. These findings suggest that CD44 isoform expression is cell type-specifically regulated in periodontium and altered according to growth phase of HGEC. PMID- 9409459 TI - Reliability of a self-reported health questionnaire in a periodontal disease study. AB - This study assesses the reliability of a self-reported health questionnaire completed by 413 subjects aged 25-74 yr in the Erie County Periodontal Disease (ECPD) Study. Specific questions on general and oral health conditions were completed by each subject during a first visit and at a follow-up examination 2 yr later, and the two compared. Results showed that the overall measure of agreement between the two visits is substantial (average kappa, kappa = 0.80). Variation by gender and age were minimal. Questions regarding allergy to medications, oral treatment, reason for tooth extraction, health symptoms and history of systemic diseases exhibited high levels of agreement (kappa ranged from 0.71-0.90). Information on vitamin and mineral intake yielded kappa = 0.63. Oral conditions scored the lowest but were still acceptable (kappa = 0.57). These findings indicate that there were no significant discrepancies in self-reported responses to the health questionnaire used in the ECPD Study. Although the information provided by the subject may not be as accurate as compared to laboratory testing, it is nevertheless a reliable source of information which can be utilized cost-effectively in research studies. PMID- 9409460 TI - A fluid-phase endocytotic capacity and intracellular degradation of a foreign protein (horseradish peroxidase) by lysosomal cysteine proteinases in the rat junctional epithelium. AB - We investigated the co-localization of lysosomal cathepsins B, H and L, and horseradish peroxidase (HRP) in junctional epithelial (JE) cells both as a fluid phase endocytotic marker to demonstrate the fluid-phase endocytotic capacity of JE cells, and to understand the morphological relationships of the endocytosed foreign substances to lysosomal cathepsins in these cells. The diaminobenzidine (DAB) histochemical and cytochemical methods and immunohistochemical avidin biotin-peroxidase complex and immunocytochemical post-embedding colloidal gold methods were used. Under light microscopy, DAB reaction products based on HRP were found in JE but were rare or absent in the oral sulcular epithelium and oral epithelium. Immunolabeling for cathepsins B and H was found in the granular structures of the cells, but no cathepsin L was identified. With electron microscopy, DAB reaction products, which indicated both HRP and the azurophil granules of neutrophils, were endocytosed into JE cells. Using a post-embedding technique, gold particles indicating HRP were present on the plasma membrane of JE cells, at the periphery of electronlucent vacuoles, and in the electrondense granules. Gold particles indicating cathepsin B or H were found in the electrondense granules. With different sizes of colloidal golds, the co localization of cathepsin B or H with HRP was indicated only in the electrondense portion of the larger vacuoles consisting of electronlucent and -dense parts. This study provided the first morphological data which indicate that JE has a fluid phase endocytotic capacity, and which suggest that the lysosomal cathepsins B and H are involved in the intracellular degradation of foreign substances invading through the gingival sulcus in JE cells. PMID- 9409461 TI - The relationship between radiographic and clinical changes in the periodontium. AB - Change in clinical attachment level (CAL) and radiographic change in crestal bone height are often used to assess periodontal breakdown and disease progression. These two variables are also used to monitor the effect of treatment. The purpose of the present longitudinal study was to evaluate the correlation between changes in CAL and alveolar bone loss. Following initial screening, 79 subjects with established periodontitis were monitored quarterly for 1 yr, using a pressure sensitive automated probe. CAL and relative attachment level (RAL) were recorded at 6 sites for each tooth. Radiographs were obtained at baseline and 1 yr. Crestal bone changes were determined using an image enhancement technique. Mean change in attachment level was 0.16 mm. Similarly, mean proximal bone loss measured radiographically was 0.16 mm. In 6.9% of all the sites, and 13.7% of all pooled interproximal sites, AL loss was in excess of the threshold defined as 2 s.d. of repeated measurements (mean 1.54 mm). Similar percentages of sites (12.9%) had radiographic evidence of proximal bone loss exceeding the threshold (0.55-1.08 mm). A site-based analysis of active sites revealed an overall poor correlation between the 2 variables (kappa value = 0.03) which was the result of a very poor sensitivity (0.16) despite a relatively good specificity (0.81). A patient-based comparison of clinical and radiographical changes revealed an overall kappa value of 0.08, with sensitivity and specificity of 0.51 and 0.56, respectively. However, baseline CAL and crestal bone height showed good correlation (r = 0.73; p = 0.0001). It is suggested that changes in CAL and radiographic bone level progress somewhat independently. Over a short-term period of time they might not follow the same course; however, in the long term, these differences seem to level off. For longitudinal monitoring of disease progression and response to therapy both methods may be needed; while for cross-sectional evaluation and long-term prospective studies, either variable may be used alone. PMID- 9409462 TI - Effects of basic fibroblast growth factor on human periodontal ligament cells. AB - In order to clarify the regulatory mechanisms of periodontal regeneration by basic fibroblast growth factor (bFGF), effects of bFGF on proliferation, alkaline phosphatase activity, calcified nodule formation and extracellular matrix synthesis of human periodontal ligament (PDL) cells were examined in this study. bFGF enhanced the proliferative responses of PDL cells in a dose-dependent manner. The maximum mitogenic effect of bFGF on PDL cells was observed at the concentration of 10 ng/ml. In contrast, bFGF inhibited the induction of alkaline phosphatase activity and the mineralized nodule formation by PDL cells. Moreover, employing the reverse transcription-polymerase chain reaction (RT-PCR) technique, we observed that the levels of laminin mRNA of human PDL cells was specifically upregulated by bFGF stimulation, but that of type I collagen mRNA was downregulated. On the other hand, the expression of type III collagen and fibronectin mRNA were not altered even when the cells were activated by bFGF. These results suggest that suppressing cytodifferentiation of PDL cells into mineralized tissue forming cells, bFGF may play a role in wound healing by inducing growth of immature PDL cells and that in turn accelerates periodontal regeneration. PMID- 9409463 TI - Prevalence of Actinobacillus actinomycetemcomitans serotypes in Japanese patients with periodontitis. AB - Oral Actinobacillus actinomycetemcomitans strains are serologically classified into 5 distinct groups, a to e. We examined the distribution of A. actinomycetemcomitans serotypes in Japanese patients with periodontitis. A total of 157 A. actinomycetemcomitans clinical isolates from diseased sites of 39 patients with periodontitis were serotyped by using serotype-specific rabbit antisera against A. actinomycetemcomitans serotypes a, b, c, d and e strains. In the immunodiffusion assay, autoclaved extracts of 42, 6, 39, 9 and 41 A. actinomycetemcomitans clinical isolates reacted with serotypes a, b, c, d and e antisera, respectively. Although 37 patients were infected with a serotype strain, 2 patients harbored 2 different serotype strains, b/e and b/untypeable. To establish a correlation between serotype and genotype of A. actinomycetemcomitans clinical isolates from 2 patients who had different serotype strains, we used arbitrarily primed polymerase chain reaction (AP-PCR) to fingerprint clinical isolates of different serotypes. The AP-PCR genotypes among 4 clinical isolates (b/e and b/untypeable) were identical to that of A. actinomycetemcomitans Y4 (serotype b), indicating the presence of multiple A. actinomycetemcomitans serotypes which are genetically homogeneous in the periodontally diseased sites of patients with periodontitis. PMID- 9409464 TI - Transforming growth factor-beta response and expression in junctional and oral gingival epithelial cells. AB - The junctional (JE) and oral gingival (OGE) epithelium show distinct morphological phenotypes and express different cell surface and keratin markers. Transforming growth factor-beta (TGF-beta) has been shown to stimulate extracellular matrix formation and inhibit proteolytic matrix degradation in periodontal wound healing. To elucidate potential roles of TGF-beta in gingival epithelial regeneration and reattachment, the present study examined the effects of TGF-beta on JE and OGE cell growth and determined the patterns of expression of mRNAs for the TGF-beta isotypes beta 1, beta 2 and beta 3 and TGF-beta receptor types I, II and III. Primary cell cultures were initiated from JE and OGE and the cell phenotypes confirmed using monoclonal antibodies to specific keratins. TGF-beta induced a significant growth inhibition in OGE cells derived from 6 different patients with a mean inhibition of 46% and a range of 16-70% (p = 0.031). Although responses varied between patients, in general maximum inhibition occurred at 10 ng/ml TGF-beta. JE cells from 5 patients showed no significant growth inhibition by TGF-beta (p = 0.125). Greater expression of TGF beta 2 and receptor type I mRNA was found in OGE than JE cells and thus appeared to be associated with differentiating epithelial cells. JE cells expressed more TGF-beta type II receptor specific mRNA than did OGE cells, but TGF-beta 1 mRNA expression was similar in JE and OGE cells. JE or OGE cultures derived from 2 of 3 patients showed expression of mRNA for the TGF-beta type III receptor. TGF-beta 3 mRNA was not detected in any of the JE or OGE samples examined. The greater sensitivity of OGE than JE to the growth inhibiting effects of TGF-beta correlated with higher expression of receptor type I mRNA which, together with the type II receptor, is required for sensitivity to growth inhibition by TGF beta. The results suggest that, in addition to structural differences, the development of functional differences in the responses of JE and OGE to TGF-beta may be associated with the formation of JE from OGE cells and the reformation of attachment after periodontal surgery. PMID- 9409465 TI - Substance P and nociceptive afferent neurones. PMID- 9409466 TI - Inactivation of a G protein-coupled inwardly rectifying K+ channel. PMID- 9409467 TI - Perinatal changes in expression of aquaporin-4 and other water and ion transporters in rat lung. AB - 1. At birth, rapid removal of lung liquid from potential airspaces is required to establish pulmonary gas exchange. To investigate the role for water channels, aquaporins (AQP) and ion transporters in this process, the mRNA expression of AQP, Na+,K(+)-ATPase and the amiloride-sensitive Na+ channel (ENaC) were studied in the fetal and postnatal rat lung. 2. The mRNA expression of all transporters studied increased postnatally. 3. The following water channels were expressed in the lung, AQP1, 4 and 5. The most specific perinatal induction pattern was observed for AQP4. A sharp and transient increase of AQP4 mRNA occurred just after birth coinciding with the time course for clearance of lung liquid. This transient induction of AQP4 mRNA at birth was lung-tissue specific. Around birth there was a moderate increase in AQP1 mRNA, which was not transient. AQP5 increased continuously until adulthood. 4. Fetal lung AQP4 mRNA was induced by both beta-adrenergic agonists and glucocorticoid hormone, which are factors that have been suggested to accelerate the clearance of lung liquid. 5. Immunocytochemistry revealed that AQP4 was located in the basolateral membranes of bronchial epithelia in newborn rats, consistent with the view that this is the major site for perinatal lung liquid absorption. 6. The Na+,K(+)-ATPase alpha 1 subunit and ENaC alpha-subunit mRNA also increased around birth, suggesting that they co-operatively facilitate lung liquid clearance at birth. 7. These data indicate that removal of lung liquid at birth is associated with pronounced and well-synchronized changes in the expression of AQP and the ion transporters studied. The transient perinatal induction of AQP4, which could be prenatally induced by beta-adrenergic agonists, and the localization of this water channel strongly suggest that it plays a critical role for removal of lung liquid at the time of birth. PMID- 9409468 TI - A C-terminal peptide of the GIRK1 subunit directly blocks the G protein-activated K+ channel (GIRK) expressed in Xenopus oocytes. AB - 1. In order to find out the functional roles of cytosolic regions of a G protein activated, inwardly rectifying potassium channel subunit we studied block of GIRK channels, expressed in Xenopus laevis oocytes, by synthetic peptides in isolated inside-out membrane patches. 2. A peptide (DS6) derived from the very end of the C-terminus of GIRK1 reversibly blocked GIRK activity with IC50 values of 7.9 +/- 2.0 or 3.5 +/- 0.5 micrograms ml-1 (corresponding to 3.7 +/- 0.9 or 1.7 +/- 0.2 mumol l-1) for GIRK1/GIRK5 or GIRK1/GIRK4 channels, respectively. 3. Dose dependency studies of GIRK activation by purified beta gamma subunits of the G protein (G beta gamma) showed that DS6 block of GIRK channels is not the result of competition of the peptide with functional GIRK channels for the available G beta gamma. 4. Burst duration of GIRK channels was reduced, whereas long closed times between bursts were markedly increased, accounting for the channel block observed. 5. Block by the DS6 peptide was slightly voltage dependent, being stronger at more negative potentials. 6. These data support the hypothesis that the distal part of the carboxy-terminus of GIRK1 is a part of the intrinsic gate that keeps GIRK channels closed in the absence of G beta gamma. PMID- 9409469 TI - Synergistic activation of guinea-pig cardiac cystic fibrosis transmembrane conductance regulator by the tyrosine kinase inhibitor genistein and cAMP. AB - 1. The regulation of cardiac Cl- current (ICl) by tyrosine and serine/threonine phosphorylation was examined in guinea-pig and rat ventricular myocytes. The protein tyrosine kinase (PTK) inhibitor genistein (GST) and phosphotyrosine phosphatase (PTP) inhibitor sodium orthovanadate (VO4) were used to modify tyrosine phosphorylation, whereas forskolin (FSK), cAMP, and other agents were used to modify cytoplasmic cAMP concentration and protein kinase A (PKA) phosphorylation. 2. Low concentrations (0.1 microM) of FSK did not activate the PKA-regulated cystic fibrosis transmembrane regulator (CFTR) ICl in guinea-pig ventricular myocytes, but strongly potentiated activation of an ICl by 20-100 microM GST. The potentiation did not occur when GST was replaced by PTK-inactive daidzein, and it was strongly inhibited by 1 mM VO4. 3. Potentiation by 0.1 microM FSK was linked to a small stimulation of the adenylate cyclase-cAMP-PKA pathway. The potentiation was not mimicked by inactive 1,9-dideoxyforskolin, and was inhibited by muscarinic stimulation (ACh) and by a PKA inhibitor. Internal application of a cAMP solution that alone was too weak to activate CFTR ICl strongly potentiated the activation of ICl by 50 microM GST and occluded potentiation by 0.1 microM FSK. 4. The foregoing suggests that potentiated ICl flows through cAMP-dependent CFTR channels. In agreement with this interpretation, GST did not increase ICl when CFTR was maximally activated by a high concentration (5 microM) of FSK and okadaic acid, and neither GST nor GST plus FSK activated an ICl in CFTR-deficient rat myocytes. The lack of effect in rat myocytes was not due to the absence of functional, channel-relevant PKA and PTK-PTP systems, because (as in guinea-pig myocytes) L-type Ca2+ current (ICa,L) was stimulated by FSK and inhibited in a VO4-reversible manner by GST. 5. The synergistic activation of CFTR by low concentrations of FSK and GST cannot be explained by either a GST-induced elevation of cAMP concentration or inhibition of serine/threonine phosphatase. Rather, it appears to be due to tyrosine dephosphorylation that facilitates PKA-mediated phosphorylation of the channels. PMID- 9409470 TI - Swelling-induced Cl- current in guinea-pig atrial myocytes: inhibition by glibenclamide. AB - 1. Whole-cell currents were recorded from guinea-pig atrial myocytes using the patch-clamp technique under conditions designed to block K+ channels, Ca2+ channels and electrogenic transporters. 2. Exposure of atrial myocytes to the hyposmotic external solution (Na+ reduction to about 70% of control) resulted in hyposmotic cell swelling which was associated with activation of an outwardly rectifying Cl- current (ICl,swell). 3. Whereas the activation of ICl,swell was not significantly affected by replacement of ATP in the pipette solution with the non-hydrolysable ATP analogue 5'-adenylyl-imidodiphosphate (AMP-PNP), its activation was greatly reduced in cells dialysed with an ATP-free pipette solution, thus indicating that the activation process of ICl,swell requires the presence of intracellular ATP, but not its hydrolysis. 4. Bath application of glibenclamide produced a concentration-dependent block of ICl,swell with a half maximal inhibitory concentration (IC50) of 60.0 microM and a Hill coefficient of 2.1. The maximal effect (100% inhibition) was obtained with 500 microM glibenclamide. The steady-state inhibition showed little voltage dependence, while glibenclamide at concentrations of more than 100 microM inhibited the outward ICl,swell more rapidly than the inward ICl,swell. The glibenclamide inhibition was fully reversible after removal of the drug, even when a maximal effect (full inhibition) was achieved at a high drug concentration (500 microM). 5. These results show that (i) glibenclamide is one of the most potent inhibitors of guinea-pig atrial ICl,swell, and (ii) atrial ICl,swell and the cystic fibrosis transmembrane conductance regulator (CFTR) Cl- currents are almost equally sensitive to inhibition by glibenclamide. PMID- 9409471 TI - Modulation of Ca(2+)-activated Cl- currents in rabbit portal vein smooth muscle by an inhibitor of mitochondrial Ca2+ uptake. AB - 1. The effects of carbonyl cyanide m-chlorophenyl hydrazone (CCCP), an inhibitor of mitochondrial Ca2+ uptake, was investigated on the properties of Ca(2+) activated chloride currents (ICl(Ca)) in rabbit portal vein smooth muscle cells using the perforated patch whole-cell voltage-clamp technique to ascertain whether this Ca2+ uptake process influences the time course of the subsarcolemmal Ca2+ signal that activates ICl(Ca). 2. In cells bathed in either physiological calcium (2 mM Cao2+) or high calcium (10 mM Cao2+) external solutions, application of CCCP (1-2 microM) evoked an inward current and prolonged the exponential decay time constant (tau) of Ca(2+)-activated Cl- 'tail' currents (Itail) evoked by Ca2+ influx through voltage-dependent calcium channels (VDCCs). The effect of CCCP on tau was greater in cells where the amplitude of Itail was relatively large and, in different cells, the effect of CCCP on tau was positively correlated with the amplitude of Itail. 3. CCCP abolished spontaneously occurring transient Ca(2+)-activated Cl- currents (STICs), but did not alter their time course before complete block. 4. Thapsigargin and cyclopiazonic acid (inhibitors of the sarcoplasmic Ca(2+)-ATPase) inhibited STICs, but did not affect the decay of Itail or STICs. 5. In conclusion, when Ca2+ enters the cell through VDCCs, the time course of the consequent Ca2+ signal in the subsarcolemmal domain containing Ca(2+)-activated chloride channels appears to be regulated by Ca2+ uptake into mitochondria. In contrast, inhibition of Ca2+ uptake by the sarcoplasmic reticulum ATPase does not seem to influence the time course of ICl(Ca). PMID- 9409472 TI - Characterization of a nicotinamide-adenine dinucleotide-dependent cation channel in the CRI-G1 rat insulinoma cell line. AB - 1. Cell-free excised membrane patches were used to examine the properties of a novel nicotinamide-adenine dinucleotide (beta-NAD+)-activated ion channel in the rat insulin-secreting cell line, CRI-G1. 2. In inside-out recordings, beta-NAD+ (0.05-1.0 mM) induced the appearance of a channel characterized by extremely slow kinetics, with mean open times in the range of seconds. The estimated EC50 for activation was 114 microM. Channel activity declined with time (run-down) following activation by beta-NAD+ in excised patches and this was not prevented by intracellular application of trypsin. 3. The single channel current-voltage relationship was linear with a conductance of 74 pS in symmetrical NaCl. The channel appears equally permeable to Na+, K+ and Cs+, exhibits an appreciable permeability to Ca2+, Mg2+ and Ba2+, but excludes anions. 4. The channel displays an unusual voltage sensitivity, with an abrupt increase in open-state probability at depolarized voltages. 5. Channel opening, in the presence of beta-NAD+, required both Ca2+ and Mg2+ to be present at the internal side of the membrane. Activation by Ca2+ required a concentration of at least 10 microM and was maximal at 0.1 mM. Ba2+ did not substitute for Ca2+ in inducing channel activity nor did it inhibit activation by Ca2+. Increasing the concentration of intracellular Mg2+ stabilized the open state of NAD(+)-activated channels. 6. The non-selective cation channel reported here differs in its gating and modulatory characteristics from non-selective cation channels described in other tissues. This channel may play a role in the pathophysiological responses of beta-cells to oxidative stress. PMID- 9409473 TI - ATP-dependent desensitization of the muscarinic K+ channel in rat atrial cells. AB - 1. Fast desensitization of the muscarinic K+ channel has been studied in excised patches from rat atrial cells. 2. In inside-out patches, ACh was present in the pipette and GTP was applied via the bath to activate the channel. In outside-out patches, GTP was present in the pipette and ACh was applied via the bath to activate the channel. In both cases, during a 30 s exposure to GTP or ACh there was a decline in channel activity as a result of fast desensitization if ATP was present. 3. In inside-out patches, fast desensitization was still observed if the muscarinic ACh receptor was bypassed and the channel was activated by GTP gamma S. This suggests that fast desensitization is a result of a modification of the channel (or the connecting G protein) and not the receptor. 4. In both inside-out and outside-out patches, channel activity was depressed and fast desensitization was reduced or absent, if ATP was not present. 5. The non-hydrolysable analogue of ATP, AMP-PNP, did not substitute for ATP in its effects on the channel. 6. The results are consistent with the hypothesis that fast desensitization of the muscarinic K+ channel is the result of a dephosphorylation of the channel. PMID- 9409474 TI - Reduced outward K+ conductances generate depolarizing after-potentials in rat supraoptic nucleus neurones. AB - 1. Whole-cell patch clamp recordings were obtained from sixty-five rat supraoptic nucleus (SON) neurones in brain slices to investigate ionic mechanisms underlying depolarizing after-potentials (DAPs). When cells were voltage clamped around -58 mV, slow inward currents mediating DAPs (IDAP), evoked by three brief depolarizing pulses, had a peak of 17 +/- 1 pA (mean +/- S.E.M.) and lasted for 2.8 +/- 0.1 s. 2. No significant differences in the amplitude and duration were observed when one to three preceding depolarizing pulses were applied, although there was a tendency for twin pulses to evoke larger IDAP than a single pulse. The IDAP was absent when membrane potentials were more negative than -70 mV. In the range -70 to -50 mV, IDAP amplitudes and durations increased as the membrane became more depolarized, with an activation threshold of -65.7 +/- 0.7 mV. 3. IDAP with normal amplitude and duration could be evoked during the decay of a preceding IDAP. As frequencies of depolarizing pulses rose from 2 to 20 Hz, the times to peak IDAP amplitude were reduced but the amplitudes and durations did not change. 4. A consistent reduction in membrane conductance during the IDAP was observed in all SON neurones tested, and averaged 34.6 +/- 3.3%. Small hyperpolarizing pulses used to measure membrane conductances appeared not to disturb major ionic mechanisms underlying IDAP, since the slope and duration of IDAP with and without test pulses were similar. 5. The IDAP had an averaged reversal potential of -87.4 +/- 1.6 mV, which was close to the K+ equilibrium potential. An elevation in [K+]o reduced or abolished the IDAP, and shifted its reversal potential toward more positive levels. Perifusion of slices with 7.5-10 mM TEA, a K+ channel blocker, reversibly suppressed the IDAP. 6. Both Na+ and Ca2+ currents failed to induce an IDAP-like current during perifusion of slices with media containing high [K+]o or TEA. However, the IDAP was abolished by replacing external Ca2+ with Co2+, or replacing 82% of external Na+ with choline or Li+. Perifusion of slices with media containing 1-2 microM TTX also reduced IDAP by 55.5 +/- 9.0%. 7. These results suggest that the generation of DAPs in SON neurones mainly involves a reduction in outward K+ current(s), which probably has little or no inactivation and can be inhibited by [Ca2+]i transients, due to Ca2+ influx during action potentials and Ca2+ release from internal stores. Na+ influx might provide a permissive influence for Ca(2+)-induced reduction of K+ conductances and/or help to raise [Ca2+]i via reverse-mode Ca(2+)-Na+ exchange. Other conductances, making minor contributions to the IDAP, may also be involved. PMID- 9409475 TI - Hypoxia-induced inhibition of calcium channels in guinea-pig taenia caeci smooth muscle cells. AB - 1. The effects of hypoxia on whole-cell current in single smooth muscle cells and on a high K(+)-induced contraction of strips of the guinea-pig taenia caeci were studied. 2. In physiological salt solution (PSS) and K(+)-based pipette solution, hypoxia (PO2 = 20 mmHg) reversibly inhibited both the inward Ca2+ current (ICa) and outward Ca(2+)-activated K+ current (IK(Ca)) components of the whole-cell current. 3. In PSS and Cs(+)-based pipette solution, hypoxia reversibly suppressed ICa by 30 +/- 5% at 0 mV. 4. When Ba2+ was used as a charge carrier, the IBa was suppressed by hypoxia in a potential-dependent manner, with the maximum of 40 +/- 7% at +10 mV. Alterations of concentrations of EGTA, GDB beta S or ATP in the pipette solution did not change the inhibitory effects of hypoxia on ICa and IBa. 5. In PSS with 2 mM CaCl2 replaced by CoCl2, hypoxia did not affect the Ca2+ influx-independent potassium current. 6. In cells voltage clamped at -20 mV hypoxia reversibly inhibited the spontaneous transient outward currents. 7. The response of high K(+)-contracted taenia caeci to hypoxia was composed of an initial rapid relaxation followed by a small transient contraction and slow relaxation. The transient contraction was blocked by atropine (1-10 microM), while relaxations were unaffected by atropine and guanethidine (10 microM). 8. The results show that hypoxia reversibly inhibits ICa and secondarily suppresses IK(Ca) due to decreased Ca2+ influx through Ca2+ channels. 9. It is suggested that inhibition of ICa was responsible for the rapid relaxation, whereas transient contraction may have been due to release of acetylcholine from nerve terminals upon hypoxia. PMID- 9409476 TI - Ca2+ current and Ca(2+)-activated chloride current in isolated smooth muscle cells of the sheep urethra. AB - 1. Isolated sheep urethral cells were studied using the perforated patch clamp technique (T = 37 degrees C). Depolarizing steps ranging from -40 to -10 mV evoked an inward current that peaked within 10 ms and a slower inward current. Stepping back to the holding potential of -80 mV evoked large inward tail currents. All three currents were abolished by nifedipine (1 microM). Substitution of external Ca2+ with Ba2+ resulted in potentiation of the fast inward current and blockade of the slow current and tails. 2. Changing the chloride equilibrium potential (ECl) from 0 to +27 mV shifted the reversal potential of the tail currents from 1 +/- 1 to 27 +/- 1 mV (number of cells, n = 5). Chloride channel blockers, niflumic acid (10 microM) and anthracene-9 carboxylic acid (9AC, 1 mM), reduced the slow current and tails suggesting that these were Ca(2+)-activated Cl- currents, ICl(Ca). 4. Caffeine (10 mM) induced currents that reversed at ECl and were blocked by niflumic acid (10 microM). 5. In current clamp mode, some cells developed spontaneous transient depolarizations (STDs) and action potentials. Short exposure to nifedipine blocked the action potentials and unmasked STDs. In contrast, 9AC and niflumic acid reduced the amplitude of the STDs and blocked the action potentials. 6. In conclusion, these cells have both L-type ICa and ICl(Ca). The former appears to be responsible for the upstroke of the action potential, while the latter may act as a pacemaker current. PMID- 9409477 TI - Thromboxane A2, released by the anti-tumour drug irinotecan, is a novel stimulator of Cl- secretion in isolated rat colon. AB - 1. A camptothecin derivative, irinotecan (Cpt-11), is a topoisomerase I inhibitor and has a strong activity against a broad range of human cancer. One of the side effects of this drug is diarrhoea. Here, we tried to determine the mediator of the irinotecan-induced Cl- secretion which may underlie this diarrhoea, using isolated mucosae of rat distal colon. 2. Irinotecan increased Cl- secretory current in a concentration-dependent manner across the mucosa, set between Ussing chambers. Thromboxane A2 (TXA2) has not been reported to date as a physiological stimulant of Cl- secretion in the distal colon. However, the major part of the present irinotecan-induced current was inhibited by selective thromboxane A2 receptor antagonists (KW-3635 and ONO-3708), and a selective thromboxane synthase inhibitor (Y-20811). In fact, we found that irinotecan stimulated the release of TXA2 in a concentration-dependent manner from the isolated mucosa into the bathing solutions. 3. Furthermore, 9,11-epithio-11,12-methano-thromboxane A2 (STA2), a stable analogue of TXA2, induced Cl- secretion, which was almost completely inhibited by the TXA2 receptor antagonists. 4. In single cells of isolated crypts, STA2 depolarized the cell and increased the membrane conductance, indicating that STA2 opened the apical Cl- channel of the crypt cells. 5. We conclude, therefore, that the irinotecan-induced endogenous TXA2 is a novel stimulant of the Cl- secretion from the crypt cells of distal colon. PMID- 9409478 TI - Local opioid inhibition and morphine dependence of supraoptic nucleus oxytocin neurones in the rat in vivo. AB - 1. Single neurones of the rat supraoptic nucleus were recorded during microdialysis of naloxone onto the ventral surface of the nucleus in anaesthetized rats. We used this combination of techniques to test whether the acute or chronic effects of systemically or centrally applied opioids upon oxytocin cell activity were due to actions of the opioids within the nucleus itself. 2. Supraoptic nucleus oxytocin neurones were identified antidromically and by an excitatory response to intravenously injected cholecystokinin. Acute intravenous injection of the kappa-agonist U50488H or the mu-agonist morphine (1 5 mg kg-1) reduced the firing rate of identified oxytocin neurones by 97.7 +/- 4.8% (n = 6) and 94.1 +/- 4.1% (n = 7), respectively. The inhibition by each of these opioids was completely reversed after administration by microdialysis (retrodialysis) of the opioid antagonist naloxone (0.1-1.0 microgram microliter-1 at 2 microliters min-1) onto the exposed ventral surface of the supraoptic nucleus. 3. Retrodialysis of naloxone (0.1-10.0 micrograms microliter-1) onto the supraoptic nucleus of rats made dependent by intracerebroventricular morphine infusion for 5 days increased the firing rate of oxytocin neurones from 0.9 +/- 0.4 to 3.1 +/- 0.7 spikes s-1 (P < 0.05, n = 6). This increase in firing rate from basal was 58.5 +/- 15.1% of that following subsequent intravenously injected naloxone (5 mg kg-1). 4. Thus, the acute inhibition of supraoptic nucleus oxytocin neurones which results from systemic administration of opioid agonists primarily occurs within the supraoptic nucleus itself, since the antagonist naloxone was effective when given into the supraoptic nucleus. Furthermore, oxytocin neurones develop morphine dependence by a mechanism which is distinct from an action on their distant afferent inputs. Nevertheless, withdrawal excitation of these afferent inputs may enhance the magnitude of oxytocin neurone withdrawal excitation. PMID- 9409479 TI - Presynaptic GABAB and adenosine A1 receptors regulate synaptic transmission to rat substantia nigra reticulata neurones. AB - 1. Patch pipettes were used to record whole-cell currents under voltage clamp in substantia nigra zona reticulata (SNR) neurones in the rat midbrain slice. Bipolar electrodes evoked synaptic currents mediated by glutamate (EPSCs) and GABAA receptors (IPSCs). 2. Baclofen reduced the amplitude of IPSCs by 48% at its IC50 value of 0.60 microM. The GABAB antagonist CGP 35348 blocked this effect with a Kd value estimated by Schild analysis of 5 microM. 3. Adenosine reduced IPSCs by 48% at its IC50 value of 56 microM. Adenosine agonists reduced IPSCs with the following rank order of potency: CPA (N6-cyclopentyladenosine) > R-PIA (R(-)N6-(2-phenylisopropyl)adenosine) > CHA (N6-cyclohexyladenosine) = NECA (5'-N ethylcarboxamidoadenosine) > 2-CADO (2-chloroadenosine) > adenosine. Schild analysis yielded a Kd value of 0.4 nM for antagonism of CPA by the adenosine A1 receptor antagonist DPCPX (8-cyclopentyl-1,3-dipropylxanthine). 4. Both baclofen and adenosine reduced the magnitude of paired-pulse depression of IPSCs, and neither blocked currents evoked by GABA, which was pressure-ejected from micropipettes. 5. Glutamate EPSCs were reduced by baclofen (IC50 = 0.78 microM) and adenosine (IC50 = 57 microM). Schild analysis yielded a Kd value of 11 microM for antagonism of baclofen-induced inhibition of EPSCs by CGP 35348. DPCPX (1 microM) completely blocked the inhibitory effects of adenosine (100 microM) and CPA (100 nM) on EPSCs. Neither adenosine nor baclofen reduced inward currents evoked by glutamate which was pressure-ejected from micropipettes. 6. These results show that presynaptic GABAB and A1 receptors reduce glutamate and GABA release from nerve terminals in the SNR. PMID- 9409481 TI - Relationship of substance P to afferent characteristics of dorsal root ganglion neurones in guinea-pig. AB - 1. The relationship between the afferent properties and substance P-like immunoreactivity (SP-LI) of L6 and S1 dorsal root ganglion (DRG) neuronal somata was examined in anaesthetized guinea-pigs. Glass pipette microelectrodes filled with fluorescent dyes were used to make intracellular recordings and to label DRG somata. The dorsal root conduction velocity (CV) and the afferent receptive properties of each unit were categorized according to criteria established in other species. Categories included a variety of low threshold mechanoreceptive classes, innocuous thermoreceptive and several nociceptive classes. Nociceptive units were further subdivided on the basis of CV and the locus of the receptive field (superficial cutaneous, deep cutaneous or subcutaneous). 2. SP-LI was determined using the avidin-biotin complex method and the relative staining intensity determined by image analysis. The possible significance of labelling intensity is discussed. Clear SP-LI appeared in twenty-nine of 117 dye-labelled neurones. All SP-LI positive units with identified receptive properties were nociceptive but not all categories of nociceptors were positive. The intensity of SP-LI labelling varied, often systematically, in relation to afferent properties. There was a tendency for nociceptive neurones with slower CVs and/or smaller cell bodies to show SP-LI. 3. Nineteen of fifty-one C fibre neurones showed SP-LI. Fewer than half the C polymodal nociceptors (CPMs) were positive. The most intensely labelled units were the deep cutaneous nociceptors and some of the CPMs in glabrous skin. C low threshold mechanoreceptors and cooling-sensitive units did not show SP-LI. 4. Ten of sixty-six A fibre neurones exhibited SP-LI, including eight of sixteen A delta nociceptors and two of fifteen A alpha/beta nociceptors. A fibre neurones exhibiting SP-LI included seven of eight deep cutaneous mechanical nociceptors and some superficial cutaneous mechano-heat nociceptors of hairy skin. In contrast, none of twenty superficial cutaneous A high threshold mechanoreceptor units or the thirty-five A fibre low threshold units (D-hair and other units) showed detectable SP-LI. 5. We conclude that SP-LI labelling in guinea-pig DRG neurones is related to (a) afferent receptive properties, (b) the tissue in which the peripheral receptive terminals are located, (c) the CV and (d) the soma size. PMID- 9409480 TI - The removal of acetylcholine by diffusion at nicotinic synapses in the rat otic ganglion. AB - 1. We have examined the clearance of synaptically released acetylcholine in the otic ganglion when acetylcholinesterase was blocked with eserine. 2. Intracellular recordings were made from otic ganglion neurones, in vitro. The decay of the excitatory postsynaptic potential (EPSP), in response to stimulation of afferent fibres, was greatly prolonged in the presence of eserine. Low frequency (0.05-4 Hz) repetitive synaptic stimulation led to a slow depolarization of the postsynaptic cell that persisted throughout the period of stimulation. This slow depolarization was blocked by the nicotinic antagonists mecamylamine (100 microM) or (+)tubocurarine (100 microM), but was unaffected by atropine (1 microM), indicating that the response was due to the activation of nicotinic receptors. 3. Following 2 Hz synaptic stimulation (30 s), the rate of rise of the slow depolarization had a time constant of 3.1 +/- 0.4 s and a peak amplitude of 12 +/- 1 mV. Upon cessation of stimulation, the depolarization decayed to resting levels with a time constant of 18.3 +/- 1.5 s (n = 23). At increasing stimulation frequencies the rate of rise of the depolarization increased. Lowering the probability of release, by adding cadmium to the perfusing solution or by lowering extracellular calcium, slowed the rise time of the response. 4. Both the onset and decay kinetics of the slow depolarization had a low temperature sensitivity, indicating that they reflect diffusional processes. 5. Repetitive stimulation (2 Hz) of the afferent nerve supplying the ganglion, in the presence of eserine, also caused a slow depolarization in cell in which we could not demonstrate a synaptic input. This indicates that synaptically released acetylcholine can spill over onto nearby neurones. 6. We conclude that at parasympathetic synapses, under physiological conditions, transmitter action is terminated by the enzymatic degradation of acetylcholine. When acetylcholinesterase is blocked, acetylcholine accumulates because its removal by diffusion is slow. PMID- 9409482 TI - Linear transduction of natural stimuli by dark-adapted and light-adapted rods of the salamander, Ambystoma tigrinum. AB - 1. We examined signal, noise and response properties of salamander rod photoreceptors by measuring: (a) the circulating current of rods which were adapted to darkness and to a wide range of backgrounds; (b) contrasts of natural environments; (c) the effect of adaptation on the linear response range of rods; and (d) the behaviour of rods responding to dynamically modulated stimuli having a range of contrasts found in nature. 2. In the dark, the circulating current contained two noise components analogous to those described in toad. A discrete noise component consisted of events occurring at a rate of 1 event per 32 s (21 degrees C) and had a variance of 0.036 pA2. A continuous noise component contributed 0.022 pA2 to the dark current, roughly equal to the discrete noise variance. 3. Exposure to a wide range of steady backgrounds (suppressing up to 80% of the circulating current), elicited a sustained fluctuating photocurrent having a power spectrum which resembled those of single photon responses and was consistent with the linear summation of single photon events; this indicates that the primary source of noise in the current is caused by the light. 4. Eighty-nine per cent of the contrasts (C) measured in natural environments had magnitude of C < 50%, where C = magnitude of I - Imean/magnitude of Imean. The linear response range elicited by brief flashes expanded with brighter backgrounds, well encompassing flash contrasts of 100%. 5. Dynamically modulated stimuli and incremental flashes having contrasts similar to those in natural scenes elicited small currents which deviated by a few picoamps about the mean and the transfer functions computed from each type of stimulus-response pair closely corresponded to one another. These results indicate that in natural environments, rods behave as linear small-signal transducers of light. PMID- 9409483 TI - Energetics of lengthening in mouse and toad skeletal muscles. AB - 1. The energetics of lengthening were studied in amphibian and mammalian skeletal muscle. The aims were to determine whether energy absorption during stretch is a general property of skeletal muscle and to investigate the influence of lengthening velocity on energy absorption. 2. Experiments were performed in vitro (21 degrees C) using bundles of muscle fibres from fast-twitch extensor digitorum longus and slow-twitch soleus muscles of the mouse and tibialis anterior muscles of a toad, Bufo marinus. Initial heat production and mechanical work done on muscles were measured during isovelocity lengthening. Enthalpy output during lengthening was calculated as the difference between the amount of heat produced and the work done. 3. For all three muscle types, more energy was put into muscles as work than was produced as heat. Thus, part of the energy put into muscles to stretch them must have been absorbed. 4. For all three muscle types, the amount of energy absorbed was constant at velocities exceeding approximately 0.5 Vmax (Vmax is the maximum shortening velocity), but was significantly lower at slow velocities of lengthening. The same amount of energy was absorbed by all three muscles when lengthened at > or = 0.5 Vmax. 5. It was concluded that absorption of energy during lengthening occurs in mammalian as well as amphibian muscle and that lengthening velocity has only a small effect on the amount of energy absorbed. PMID- 9409484 TI - Mechanism of action of endothelin in rat cardiac muscle: cross-bridge kinetics and myosin light chain phosphorylation. AB - 1. The molecular mechanism of inotropic action of endothelin was investigated in rat ventricular muscle by studying its effects on characteristics of isometric twitch, barium-induced steady contracture and the level of incorporation of 32Pi into myosin light chain 2. 2. Exposure of rat papillary muscle to endothelin caused an increase in isometric twitch force but did not alter the twitch-time parameters. 3. Endothelin did not significantly change the maximum contracture tension but did cause an increase in contracture tension at submaximal levels of activation, without changes in the tension-to-stiffness ratio and kinetics of attached cross-bridges. Kinetics of attached cross-bridges were deduced during steady contracture from complex-stiffness values, and in particular from the frequency at which muscle stiffness assumes a minimum value, fmin. Endothelin did not alter fmin. 4. Endothelin caused an increase in the level of incorporation of 32Pi into myosin light chain 2 without a concurrent change in the level of incorporation of 32Pi into troponin I. 5. We conclude that the inotropic action of endothelin is not due to an increase in the kinetics of attached cross bridges, nor due to a change in the force per unit cross-bridge, but may result from an increased divalent cation sensitivity caused by elevated myosin light chain 2 phosphorylation, resembling post-tetanic potentiation in fast skeletal muscle fibres. PMID- 9409486 TI - Development of electrical rhythmicity in the murine gastrointestinal tract is specifically encoded in the tunica muscularis. AB - 1. Interstitial cells of Cajal (ICCs) have been identified as pacemaker cells in the gastrointestinal (GI) tracts of vertebrates. We have studied the development of ICCs in pacemaker regions and the onset of electrical rhythmicity in the gastric antrum, small bowel and proximal colon of the mouse. 2. ICCs, as detected by c-Kit immunofluorescence, were found during embryogenesis in regions of the GI tract that eventually become pacemaker areas. Prior to birth, these cells were organized into well-structured networks, and by the end of the embryonic period the morphology of ICC networks in pacemaker regions appeared very similar to that observed in adult animals. 3. Electrical rhythmicity was recorded prior to birth (by E18) in the proximal GI tract (stomach and jejunum), and this activity developed to adult-like behaviour within a week after birth. In the ileum and proximal colon rhythmicity developed after birth, and adult-like characteristics were apparent within the first week. 4. Post-junctional responses of smooth muscles to neural inputs could be recorded at birth, and stimulation of intrinsic nerves often led to oscillatory activity resembling slow waves for up to several minutes following brief stimuli. Nerve stimulation augmented spontaneous activity in the proximal portions of the GI tract and elicited rhythmic activity temporarily in quiescent tissues of the distal GI tract. 5. ICCs and rhythmicity developed in an apparently normal manner in tissues isolated at birth and placed in organ culture. These data suggest that the tunica muscularis provides a suitable microenvironment for the development of ICCs and rhythmicity without the need for extrinsic stimuli. 6. Treatment of small intestinal tissues taken from embryos at E15 with neutralizing c-Kit antibodies abolished ICC development and the organization of ICCs into networks that typically occurs during the late embryonic period. Treatment of muscles taken from newborn animals with c-Kit antibodies blocked postnatal development of ICCs, disrupted already established and functional ICC networks, and rendered muscles electrically quiescent. 7. In summary, ICC networks develop in the pacemaker regions of the murine GI tract before birth. Development and organization of ICCs of the myenteric plexus region into networks precedes the development of electrical rhythmicity. Post-natal development of electrical rhythmicity is mainly characterized by enhancement of the amplitude and frequency of slow waves. The development of ICCs and electrical rhythmicity persists in vitro. ICCs appear to be necessary for the initiation of electrical rhythmicity. These findings provide further evidence for the pacemaker role of ICCs. PMID- 9409488 TI - Telemedicine and the doctor-patient relationship. PMID- 9409485 TI - Consequences of metabolic inhibition in smooth muscle isolated from guinea-pig stomach. AB - 1. In smooth muscle isolated from the guinea-pig stomach, cyanide (CN) and iodoacetic acid (IAA) were applied to block oxidative phosphorylation and glycolysis, respectively. Effects of IAA on generation of spontaneous mechanical and electrical activities were systematically investigated by comparing those of CN. Spontaneous activity ceased in 10-20 min during applications of 1 mM IAA. On the other hand, application of 1 mM CN also reduced the spontaneous activity, but never terminated it. In the presence of CN the negativity of the resting membrane potential was slightly reduced. 2. When spontaneous activity ceased with IAA, the resting membrane potential was not significantly affected. Also, before ceasing, the amplitude and duration of the spontaneous electrical activity were significantly reduced. The amplitude of the electrotonic potential was, however, not changed by IAA. Further, glibenclamide did not prevent the effects of IAA. These results suggest that, unlike cardiac muscle, activation of metabolism dependent K+ channels in stomach smooth muscle does not seem to play a major role in reducing and terminating spontaneous activity during metabolic inhibition. 3. Carbachol-induced contraction transiently increased, and subsequently decreased gradually during application of IAA. 4. After 50 min application of IAA, when there was no spontaneous activity, the concentrations of phosphocreatine (PCr) and ATP measured with 31P nuclear magnetic resonance decreased to 60 and 80% of the control, respectively, while inorganic phosphate (Pi) concentration paradoxically fell to below detectable levels. During subsequent prolonged application of IAA, high-energy phosphates steadily decreased. On the other hand, after 50 min CN application, [PCr] and [ATP] decreased to approximately 30 and 80% of the control, respectively, while [Pi] increased by 2.6-fold. 5. In the presence of either CN or IAA, spontaneous mechanical and electrical activities were reduced or eliminated, although amounts of high-energy phosphates sufficient to contract smooth muscle remained. It can be postulated that some mechanism(s) related to energy metabolism, but not including ATP-sensitive K+ channels, plays an important role in generating spontaneous activity in guinea-pig stomach smooth muscle. During metabolic inhibition the energy metabolism-dependent mechanism(s) would preserve high-energy phosphates, and consequently cell viability, by stopping spontaneous activity. PMID- 9409487 TI - Mental rehearsal of motor tasks recruits alpha-motoneurones but fails to recruit human fusimotor neurones selectively. AB - 1. As mental rehearsal of movements activates multiple cortical areas associated with movement, we assessed whether this increases fusimotor drive and whether enhanced muscle spindle activity could contribute to the improvement in skill that accompanies mental rehearsal. 2. Microneurographic recordings were made from six muscle spindle afferents innervating extensor muscles in the forearm or tibialis anterior, which were selected because their discharge increased during very weak contractions. Activity was monitored while subjects imagined performing a range of activities including simple and complex movements involving the relevant muscles. 3. No activation of muscle spindle afferents occurred during imagined motor tasks without EMG. When the relevant muscles contracted during mental rehearsal, spindle discharge increased, much as in weak contractions. 4. Mental rehearsal increased background EMG in the involved muscles and also increased H reflex amplitude independently of EMG changes. 5. Although there was no evidence for selective fusimotor activation during imagined movement, skeletomotor activity and reflex excitability increased. Similar changes occur with preparation for movement following a cue. It is likely that mental rehearsal usually involves unintentional performance of the planned motor task. PMID- 9409489 TI - Update on viral hepatitis. PMID- 9409490 TI - Acute liver failure. PMID- 9409491 TI - Investigation and management of gastrointestinal motility disease. PMID- 9409492 TI - Coeliac disease. AB - Coeliac disease, or gluten sensitive enteropathy is a common disorder and results from exposure to gluten in the diet of genetically susceptible individuals. Environmental factors may influence both the age of presentation and the severity of symptoms. Screening by quantifying anti-gliadin and anti-endomysial antibody titres and diagnosis by small intestinal biopsy are both straightforward. A gluten free diet produces clinical and symptomatic improvement and decreases the rate of complications, including gastrointestinal malignancy. Current research is likely to improve our understanding of the disease pathogenesis, the structure of the toxic cereal peptides, and the genetics of the condition. PMID- 9409493 TI - Diagnostic dilemmas in colitis. PMID- 9409494 TI - Clinical genetic services: activity, outcome, effectiveness and quality. Summary of a report of the Clinical Genetics Committee of The Royal College of Physicians. PMID- 9409495 TI - Pituitary tumours: recommendations for service provision and guidelines for management of patients. Summary of a consensus statement of a working party from the Endocrinology and Diabetes Committee of The Royal College of Physicians and the Society for Endocrinology in conjunction with the Research Unit of the Royal College of Physicians. PMID- 9409496 TI - Tuberculosis--an epidemic of injustice. PMID- 9409497 TI - Financing the rising cost of haemophilia care at a large comprehensive care centre. AB - Haemophilia affects 1 in every 6,000 males. Patients with haemophilia A receive treatment with factor VIII (FVIII) and those with haemophilia B receive factor IX (FIX). In the UK, patients receive their treatment from comprehensive care centres (CCCs) or haemophilia centres. Over the last two decades the amount of clotting factor used per patient has increased; the quality of the clotting factors available and the methods of administration have also improved. As a consequence, the cost of providing care has increased substantially. In theory, the nature and level of haemophilia treatment is specified in contracts between purchasers and providers, ensuring that the costs of treating patients are fully recovered. However, at our large CCC, which has 1,700 registered patients with inherited bleeding disorders, the costs of care regularly exceed contract revenue. This paper describes the cost pressures and difficulties faced by a North London Trust in an attempt to maintain, and in some instances improve, the services provided within its CCC. PMID- 9409498 TI - Local anaesthetic infiltration prior to arterial puncture for blood gas analysis: a survey of current practice and a randomised double blind placebo controlled trial. AB - BACKGROUND: Infiltration with local anaesthetic (LA) is recommended before arterial puncture (AP) for blood gas analysis. A telephone survey of 100 junior hospital doctors established that 84% never used LA; the reason for this cited by 47% of doctors was that they considered the injection to be as painful as AP itself. We therefore undertook a randomised double blind placebo controlled trial to establish whether the recommendation for LA is justified. METHODS: Patients undergoing AP were randomly allocated to one of three groups before arterial stab: A--Infiltration with 2% lignocaine; B--Infiltration with normal saline; C- No infiltration. Patient and doctor then rated the discomfort of the procedure. RESULTS: Both patients and doctors rated the pain of AP less when LA was used. Infiltration with LA was no more painful than with placebo. AP was no more difficult following LA as assessed by passes made, times skin broken and the doctor's rating of the procedure. CONCLUSION: This study supports the recommendation that LA should be infiltrated before radial artery puncture is performed. The belief that the use of LA makes the procedure more difficult and is as painful as arterial puncture should be dispelled. PMID- 9409499 TI - Junior doctors and clinical audit. AB - OBJECTIVES: To assess the extent of junior doctor involvement in clinical audit, the degree of support from audit staff, and the perceived value of the resulting audits. DESIGN: Postal survey of National Health Service (NHS) junior doctors. SUBJECTS AND SETTINGS: 704 junior doctors in central Leeds hospitals, June 1996. RESULTS: Questionnaires were returned by 232 respondents (33%), 211 (31%) were completed; 157 respondents (74%) had personally performed audit. Mean (+/- SD) duration since last audit project was 14.9 (14.1) (range 0-84) months. Of the respondents who had personally performed audit, 88 (56%) did not use the hospital audit department, 60 (38%) received no guidance and only 19 (12%) were involved in re-auditing the same project. Mean (+/- SD) time spent per audit project was 27.8 (37.7), (range 2-212) hours. Seventy-five junior doctors (48%) were aware of subsequent change in clinical practice, 41 (26%) perceived a negative personal benefit from audit, 33 (21%) perceived a negative departmental benefit, and 42 (27%) felt that audit was a waste of time. CONCLUSIONS: A large proportion of junior doctors are involved in audit projects that do not conform to established good practice and which have a low impact on clinical behaviour. Although junior doctors feel that there is inadequate assistance and poor supervision whilst performing audit, they still support the principle of audit. There is a need to improve the quality and supervision of audit projects performed by junior doctors. PMID- 9409500 TI - The provision of CME meetings for physicians. AB - 1. A large number of meetings on a wide variety of topics are available for a majority of consultant physicians. 2. Many meetings consist of lectures or presentations; more diverse and participatory learning methods are required. 3. The pharmaceutical industry provides sponsorship for 45% of all meetings, much more in some medical specialties. 4. It should be possible and easy to obtain information about approved educational events from a WEB page. PMID- 9409501 TI - Hypoxaemia and supplemental oxygen therapy in the first 24 hours after myocardial infarction: the role of pulse oximetry. AB - OBJECTIVE: To assess the incidence and degree of hypoxaemia in patients with acute myocardial infarction and evaluate the nation-wide perception and usage of oxygen therapy. DESIGN: Postal survey of all coronary care units (CCU) in England of their use of prescribed oxygen and pulse oximetry. Prospective randomised study of 50 patients presenting within 24 hours of onset of myocardial infarction, half of whom received oxygen therapy. Oxygen saturation (SpO2) as continuously measured by pulse oximetry, and arrhythmias and ST segment changes were recorded on simultaneous 24-hour ambulatory Holter monitors. RESULTS: In 53% of UK coronary care units oxygen is not routinely prescribed but in only 3% is a pulse oximeter used to aid management. In patients presenting with acute myocardial infarction the incidence of hypoxaemia (SpO2 < 90%) was 70% and severe hypoxaemia 35% in those not given oxygen, compared with only 27% and 4% in patients given oxygen therapy. The only patient to receive oxygen on clinical grounds had an oxygen saturation of 71%. Severe hypoxaemia (SpO2 < 80%) occurred significantly less often (1 and 7 patients, p < 0.05) in patients given oxygen. There were no differences in arrhythmias or ST segment changes between groups. CONCLUSION: Hypoxaemia occurs frequently in patients in the first 24 hours after acute myocardial infarction. It is effectively and easily treated with supplemental oxygen which can be guided by pulse oximetry. This is rarely done. Measurements of oxygen saturation are therefore justified in all patients to guide oxygen therapy unless there is a decision to give all patients supplemental oxygen: this we believe to be unnecessary. PMID- 9409502 TI - The management of pulmonary tuberculosis notified in England and Wales in 1993. AB - We have compared the management of 925 cases of pulmonary tuberculosis reported to the 1993 national tuberculosis notification survey with the recommended standards of treatment. Forty-eight per cent of patients were white, 36% came from the Indian subcontinent (ISC) and 15% were of other ethnic origin. Most patients (86%) were under the care of thoracic physicians. Sputum microscopy was positive in 44%, and culture confirmation was obtained in 64% of cases. Drug resistance was reported in 30/582 isolates (5%), ranging from 13% in Black Africans to 4.5% in ISC ethnic groups and 2% in the whites, with none reported in those of Black-Caribbean origin. Almost all patients (94.5%) were started on a recommended drug combination, but only 74% continued to receive one, with thoracic physicians significantly more likely than other physicians to use a recommended combination. Non-standard durations of either initial and/or continuation phase therapy were used in 303 patients, but in only 167 was a satisfactory reason given for the modification. Definite or suspected drug toxicity was reported in 79 (9%) and was significantly more likely with non standard regimens. Seventy-two patients died before the survey was carried out one year after their notification, only 15 of them directly due to tuberculosis. Of the 815 cases observed to treatment completion, 430 (53%) were then discharged. There were adequate reasons for follow-up after the end of treatment in all but 98 of those so managed. Although the results were satisfactory overall, continued efforts are required to increase the percentage of patients treated with evidence-based recommended regimens and durations of chemotherapy. PMID- 9409504 TI - Genetics and the future of human longevity. PMID- 9409503 TI - The management of lymph node tuberculosis notified in England and Wales in 1993. AB - We have compared the management of 219 cases of lymph node tuberculosis reported to the 1993 national notification survey with the recommended standards of treatment. The diagnosis was supported by positive histology and bacteriology in 81 cases (37%), positive histology in 70 (32%), positive bacteriology in 26 (12%), and on only clinical grounds in 40 (18%). Most patients (88%) were under the care of thoracic physicians. Almost all (97%) were commenced on a recommended drug combination, but only 81% continued to receive it, with thoracic physicians more likely than other physicians to use a recommended combination. Non-standard durations of the initial and/or continuation phases of treatment were used in 83 patients, but in only 49 cases was a satisfactory reason given for the modification. Definite or suspected drug toxicity was reported in 22 cases (10%), and was significantly more likely with non-standard regimens. There were no deaths. Of the 209 patients observed to treatment completion, 129 (62%) were then discharged. There were adequate reasons for follow-up after the end of treatment in all but 32 (15%) of those so managed. Further education is required to increase the percentage of patients treated with evidence-based regimens and durations of chemotherapy. PMID- 9409505 TI - A correct compassion: the medical response to an ageing society. The Harveian Oration of 1997. PMID- 9409506 TI - Medics, lawyers and the courts. PMID- 9409508 TI - Cardiological examinations. PMID- 9409507 TI - Wegener's granulomatosis from infancy to adolescence. PMID- 9409509 TI - A filter immunobinding technique for the rapid detection and simultaneous identification of avian and bovine mycoplasmas. AB - A filter immunobinding (FIB) method was developed for the detection and identification of mycoplasmas. Type strains of a total of 18 avian and bovine mycoplasma species propagated in broth media were diluted and immobilized on a nitrocellulose membrane as antigens for investigating the specificity with rabbit hyperimmune sera. Non-specific FIB reactions were easily eliminated by the procedure of absorbing rabbit hyperimmune sera in the broth. Absorbed rabbit hyperimmune sera exhibited clear species-specificity with mycoplasma antigens by the FIB. These specific reactions always agreed with the results of identification by tests of biochemical properties and growth inhibition for the isolates of M. bovirhinis, M. bovis, M. columbinum, M. columborale, M. gallisepticum and M. synoviae. Some bovine mycoplasma species, which were impossible to identify by growth inhibition test, because of their strong production of film and spots on the agar, specifically reacted with absorbed rabbit hyperimmune sera against M. bovis in the FIB. The detection limit of mycoplasmas by this method was about 10(4)-10(5) colony-forming units/ml, which is lower than that of colony determination on agar. The FIB seems to be a useful technique for rapid detection and simultaneous identification of mycoplasmas. PMID- 9409510 TI - Innervation of two peptidergic (substance P and calcitonin gene-related peptide) nerves in the cerebral arteries and choroid plexus of the Japanese Newt (Triturus pyrrhogaster). AB - The pattern of cerebrovascular substance P (SP) and calcitonin gene-related peptide (CGRP) immunoreactive (-IR) innervation was investigated in the newt. SP IR nerves supplying the cerebral arterial tree and choroid plexus were positive for CGRP, but negative for vasoactive intestinal polypeptide or neuropeptide Y. It is suggested that cerebrovascular SP- and CGRP-IR axons are sensory in nature. The supply of SP- and CGRP-IR nerves to the major cerebral arteries is relatively poor. Nevertheless, numerous SP- and CGRP-IR axons, which are contained in the fiber bundles on the cerebral carotid artery and the basilar artery, spread widely over the microvascular-epithelial regions of the choroid plexuses. It must be considered in relation to the significant role of SP- and CGRP-IR neuronal mechanisms responsible to the microcirculation, cerebrospinal fluid (CSF) production and transport action within the choroid plexus in the nutrition of the newt brain via the CSF. PMID- 9409511 TI - Effect of attenuated Erysipelothrix rhusiopathiae vaccine in pigs infected with porcine reproductive respiratory syndrome virus. AB - Twenty 2nd specific pathogen-free pigs were divided into 4 groups: Group A were infected with porcine reproductive and respiratory syndrome (PRRS) virus at 6 weeks of age and treated with available swine erysipelas and swine fever combined vaccine (vaccinated) at 7 weeks of age; Group B were vaccinated at 7 weeks of age and infected with PRRS virus at 8 weeks of age; Group C were vaccinated at 7 weeks of age: Group D were neither vaccinated nor infected with PRRS virus. All pigs were challenged to Erysipelothrix rhusiopathiae C42 strain at 10 weeks of age. No clinical signs appeared after vaccination of group A and B pigs, thus confirming that the safety of the vaccine was not influenced by infection with PRRS virus. None of the pigs in Groups A and C developed erysipelas after challenge exposure to E. rhusiopathiae. In contrast, fever and/or urticaria appeared transiently in all pigs of Group B after challenge exposure. At the time of challenge exposure to E. rhusiopathiae, the PRRS virus titer was high in sera of Group B, but was low in those from Group A. However, vaccination of pigs with attenuated E. rhusiopathiae was effective in dual infection with PRRS virus and E. rhusiopathiae, because the clinical signs were milder and the E. rhusiopathiae strain was less recovered from these pigs compared to pigs of group D. PMID- 9409512 TI - Expression pattern of the mitochondrial capsule selenoprotein mRNA in the mouse testis after puberty; in situ hybridization study. AB - Mitochondrial capsule selenoprotein (MCS) has been known as a structural protein of the mitochondrial sheath in spermatozoa. In this study, to determine the expression pattern of MCS mRNA in the mouse testis after puberty, in situ hybridization using digoxigenin-labeled RNA probes for MCS was performed in the testes of 8- and 20-week-old ICR mice. In the testes of both ages, MCS mRNA first appeared in step 3 round spermatids, gradually increased during early spermiogenesis, and persisted a high level until step 14 spermatids. After the step 14 spermatids, the signal began to decline and was weakly detected in steps 15-16 spermatids. On the other hand, compared with that in the testes of 8-week old mice, MCS mRNA level in the testes of 20-week-old mice increased over 2-fold at stages VI-III, while it slightly increased at stages IV-V. These findings suggest that MCS gene transcription may be up-regulated after puberty in the mouse testis. PMID- 9409513 TI - A clinical evaluation of blood pressure through non-invasive measurement using the oscillometric procedure in conscious dogs. AB - The oscillometric procedure was used to measure the heart rate as well as the systolic, mean, and diastolic blood pressures of 152 dogs (102 in the control group, 13 in the group with renal disease, 37 in the group with heart disease) who were brought to Azabu University of Veterinary Teaching Hospital. It was demonstrated that the blood pressure and heart rate of the control group lowered and tended to become stable as the number of measurements increased. No appreciable difference was identified in the measurements of either blood pressure or heart rate in the forelimbs and tail head. With regard to gender, males showed a significantly higher value than females (p < 0.05). No interaction was identified between age and blood pressure. No difference was identified in the heart rate in all groups. In an investigation of blood pressure in all groups, the renal disease group showed significantly higher values (p < 0.05) than either the control or the heart disease group in all values of systolic, mean and diastolic pressures. These results indicated that dogs with renal disease can manifest hypertension. PMID- 9409514 TI - Inhibitory effects of glucocorticoids on proliferation of canine mast cell tumor. AB - The inhibitory effect of glucocorticoids (GCs) on proliferation of canine mast cell tumor (MCT) was studied using two types of MCT cells; JuMC cells and LuMC cells derived from spontaneous canine cutaneous and intestinal MCT, respectively. In in vitro study, growth of JuMC cells was significantly inhibited with more than 1 nM GCs and apoptotic-like cell death was seen, while that of LuMC cells was never inhibited even with 10 microM GCs. Growth rate of masses in nude mice developed by inoculation of JuMC cells was reduced in a dose-dependent manner by administration of GC, while growth inhibition of masses developed by inoculation of LuMC cells was minimal with increasing GC doses. Competitive binding studies and Scatchard analysis demonstrated the presence of high-affinity, low capacity GC receptors in both JuMC and LuMC cells. Kd was estimated to be 1.30 nM in JuMC cells and 0.45 nM in LuMC cells, respectively. It is concluded that canine cutaneous MCT cells responded to GCs in vitro and in vivo, whereas intestinal MCT cells did not, though both types of cells had specific GC receptors. PMID- 9409515 TI - Characterization of canine herpesvirus glycoprotein D (hemagglutinin). AB - Glycoprotein D (gD) of canine herpesvirus (CHV) YP2 strain was expressed in COS-7 and insect (Spodoptera frugiperda; Sf9) cells. The gDs expressed in COS-7 and Sf9 cells reacted with a panel of monoclonal antibodies (MAbs) against CHV gD (hemagglutinin) and an MAb 25C9 against feline herpesvirus type 1 (FHV-1) gD by indirect immunofluorescence assay, and possessed a molecular weight (MW) of approximately 51-55 and 41-46 kilodalton (kDa), respectively, when examined by immunoblot analysis. After treatment with tunicamycin, the MW of the gD expressed in Sf9 cells became approximately 37 kDa. By hemadsorption (HAD) tests using canine or feline red blood cells (RBC), COS-7 cells expressing CHV gD adsorbed only canine RBC, but not feline RBC, whereas control COS-7 cells expressing FHV-1 gD adsorbed feline RBC, but not canine RBC. By hemagglutination (HA) tests, lysates of Sf9 cells expressing CHV gD agglutinated canine RBC, but not feline RBC. These HA and HAD activities were inhibited by HA-inhibition MAbs against CHV gD. Control lysates of Sf9 cells expressing FHV-1 gD agglutinated only feline RBC. Serum from mice inoculated with lysates of Sf9 cells expressing CHV gD possessed a high titer of virus-neutralizing activities against CHV infection. These results indicated that CHV gD is structurally similar to FHV-1 gD, but is functionally different from FHV-1 gD. PMID- 9409516 TI - Analysis of distribution of Campylobacter jejuni and Campylobacter coli in broilers by using restriction fragment length polymorphism of flagellin gene. AB - The incidence of Campylobacter jejuni and Campylobacter coli in broiler farms was 33.9% (19/56). C. jejuni-positive flocks accounted for 20.0% (17/85) and C. coli positive ones was 4.7% (4/85). There were 14 patterns (fla type) of restriction fragment length polymorphism (RFLP) of flagellin A gene among these 22 strains of C. jejuni and C. coli including the standard strain C. jejuni ATCC 33560. Different fla types of Campylobacter were isolated from broilers in different growing cycles on the same farms. Four strains of C. jejuni were isolated from four breeder farms and four fla types of C. jejuni were detected from their progenies reared on growing farms. Three fla types of C. jejuni detected from the progenies were different from those of each breeder. Also, the other three fla types of C. jejuni were detected from different progenies of each growing farm during the next growing cycle. These findings indicate that the RFLP analysis may contribute to epidemiological studies of C. jejuni and C. coli contamination of broilers and suggest the risk of contamination with different types of Campylobacter in every growing cycle of broilers on the farm even on the same farm. They also supported that there was little likeliness of the vertical transmission of C. jejuni and C. coli from breeders to broilers. PMID- 9409517 TI - Effect of dietary zinc content on 65Zn metabolism in mice. AB - 65Zn is one of the induced radioactive nuclides which are generated in power reactors. In the present experiment, several parameters of 65Zn metabolism were studied in mice maintained on diets with various zinc contents from 45 to 4,500 mg/kg to evaluate the efficacy of the dilution method for radiation protection against internal contamination with 65Zn. Gastrointestinal absorption of 65Zn was suppressed and its excretion accelerated as the dietary zinc content increased over a wide range. Clearance of 65Zn from tissues was generally accelerated by feeding mice a high-zinc diet, but that from the femurs was not affected by dietary zinc content. Zinc concentrations in tissues were regulated homeostatically up to a dietary zinc content of 1,350 mg/kg. Although a significant accumulation of zinc occurred in the liver, pancreas, kidneys, and femurs when mice were given 4,500 mg/kg diet, the concentrations except in the femurs recovered within a few days after switching to a normal-zinc diet. These results suggest that oral administration of zinc is effective for preventing the absorption and for enhancing the excretion of 65Zn to protect the body from internal radiation exposure with this isotope. PMID- 9409518 TI - Enhancement of passive immunity with maternal vaccine against newborn calf diarrhea. AB - The effects of a maternal vaccine against newborn calf diarrhea associated with group A bovine rotavirus (BRV), bovine coronavirus (BCV), bovine parvovirus and K99 Escherichia coli (E. coli) were examined on a beef cow-calf herd. After vaccination, serum or colostrum antibody titers to BRV, BCV and E. coli K99 in the vaccinated cows were significantly higher than those in unvaccinated control cows. Serum antibody titers to BRV, BCV and E. coli K99 in calves from the vaccinated cows were also significantly higher than those in calves from the control cows for 3-4 weeks after birth. These results suggested that the immunization of cows with the maternal vaccine enhanced the passive immunity levels in calves against BRV, BCV and K99 E. coli. PMID- 9409519 TI - Pelvic aneurysmal bone cyst in a dog. AB - A three-year-old male Siberian Husky dog was referred to the Veterinary Teaching Hospital in Osaka Prefecture University with a complaint of difficulty in expelling the stools. By rectal examination, a mass as big as a fist could be detected occupying the cavum pelvis. Radiographically the mass had a thin bony shell bulging from the pubic periosteum. In the shell, radiolucent trabeculation gave the area a "soap bubble" appearance. The cut surface of the removed mass showed a honeycomb-like pattern constituted of some small loculate bony cysts. These cysts were separated from each other by a fibrous or bony trabeculae with blood-filled vascular channels or sponge-like structures. From clinical and pathological findings, this mass was diagnosed as a pelvic aneurysmal bone cyst. After surgery, the patient completely recovered without tenesmus. PMID- 9409520 TI - Isolation of Streptococcus equi subsp. equi from thoroughbred horses in a racehorse-breeding area of Japan. AB - For determination whether strangles has invaded the Hidaka district of Hokkaido, the main racehorse-breeding area of Japan, a epizootiological survey with bacterial isolation was carried out during the breeding season in 1995. Streptococcus equi subsp. equi, which is the causative agent of strangles, was isolated from two Thoroughbred horses with submandibular lymphadenitis. Isolates were identified by serological grouping, biochemical tests and analysis of cell surface proteins by Western immunoblotting. Through this survey, it revealed that S. equi subsp. equi has invaded the Hidaka district and that strangles has become prevalent in racehorse-breeding farms in this area. PMID- 9409521 TI - Susceptibility of several species of Cyprinidae and Salmonidae freshwater fish to larval Gnathostoma nipponicum infection. AB - Susceptibility of five species of Cyprinidae and Salmonidae freshwater fish to the early third-stage larvae (EaL3) and advanced third-stage larvae (AdL3) of Gnathostoma nipponicum infection were examined. Two fish species inoculated orally with EaL3 were infected, and AdL3 were recovered from them with rate of 21.0% in Tribolodon hakonensis and 0.5% in Cyprinus carpio at 30 days postinoculation (PI). Attempts to infect five fish species with AdL3 were all successful. The recovery rate of AdL3 was 69.0% in T. hakonensis, 47.5% in Carassius auratus subsp., 35.0% in C. carpio, 53.0% in Oncorhynchus masou, and 32.0% in O. mykiss at 10 days PI. These results confirmed that the Cyprinidae and Salmonidae fish species reported here were susceptible to larval G. nipponicum infection and AdL3 had higher infectivity to them than the EaL3. PMID- 9409522 TI - Role of host immune response in the occurrence of gastropathy in rats infected with larval Taenia taeniaeformis. AB - Decrease of spleen weight from peak to control levels together with a corresponding decline of serum IgG level at the onset of gastric hyperplasia in Taenia taeniaeformis-infected euthymic rats, may indicate that a form of down regulation of host immune response during the course of larval T. taeniaeformis infection could facilitate the occurrence of gastropathy in rats. Gastric hyperplasia developed in T. taeniaeformis-infected athymic nude rats, indicating that occurrence of gastropathy associated with larval T. taeniaeformis infection in the rat is T cell independent. Apparently, gastric hyperplasia appeared early in nude rats which suggests that absence of T cell-mediated response could have facilitated its early occurrence. PMID- 9409523 TI - Vaginal atresia with transverse septum in a cat. AB - A six and half-year-old nulliparous mixed breed cat which had the complaints of vomiting, abdominal distention and depression was presented to the Veterinary Teaching Hospital of Osaka Prefecture University. She was suspected of pyometra by clinical signs and tests. By laparotomy, it was clarified that both uterine horns and vagina showed distension by the accumulation of secretions, and the vagina ended blindly leaving a tough connective tissue at the border between cranial and caudal part of the vagina. Postoperative contrast-radiograph of the remaining vagina proved it had no persistence of the hymen. From these findings, the condition was diagnosed as a feline atresia vaginalis with the transverse vaginal septum which is caused by the embryonic failure of canalization of the paramesonephric duct between the end of the Mullerian duct and the urogenital sinus. PMID- 9409524 TI - Apoptosis enhanced by soluble factor produced in feline immunodeficiency virus infection. AB - Feline immunodeficiency virus (FIV)-infected cells have been shown to undergo apoptosis by treatment with tumor necrosis factor alpha (TNF-alpha). This study detected a soluble factor which enhanced the apoptosis induced by TNF-alpha treatment. The sensitivity to TNF-alpha in the induction of apoptosis in a feline fibroblastoid cell line (CRFK) cells was significantly enhanced when the culture supernatant of FIV-infected CRFK cells or plasma samples from FIV-infected cats was added to the culture. These findings suggested that FIV infection induces production of a soluble factor which enhances CRFK cells sensitivity to TNF-alpha induction of apoptosis both in vitro and in vivo. This factor may contribute to the loss of lymphocytes in cats infected with FIV. PMID- 9409525 TI - Effects of epidermal growth factor on maternal and fetal serum amino acid levels in rats. AB - Pregnant rats were subcutaneously administered with mouse epidermal growth factor (EGF) at the concentration of 0, 100, or 200 micrograms/kg body weight/day from day 18 to 21 of gestation. The amino acid analysis by high-performance liquid chromatography demonstrated that the umbilical venous/maternal and fetal/maternal ratio of serum proline concentration increased in EGF dose-dependent manner accompanied by the increase in the ratios of total fetal weight and placental weight to maternal body weight gain. These results suggested that EGF regulates fetal growth by, as one of its possible mechanism, promoting placental proline supply from mother to fetus. PMID- 9409526 TI - Hepatic oxygen supply, energy charge, and histological findings in dogs with portal vein arterialization. AB - Hepatic oxygen supply, energy charge (EC), and histology were examined comparatively in dogs with portal vein anastomosis (PVA group), and PA in addition to PVA (PA group). The PVA group showed a lower level of hepatic oxygen supply than those of the PA group throughout the experimental period, and also showed decreases of adenosine triphosphate (ATP) and EC level after blood perfusion. In contrast, the oxygen supply and consumption were stable in the PA group. A temporary fall of ATP level was followed by recovery to the preperfusion level in the PA group. Histological examination indicated the collapse of hepatic cords with granular and vacuolar degeneration in only the PVA group. These findings suggested that PA, when supplemented to PVA, is an available technique for preventing hepatic failure caused by ischemic conditions. PMID- 9409527 TI - Pathology of interdigital glands in a wild Japanese serow (Capricornis crispus) infected with parapoxvirus. AB - The interdigital glands of a Japanese serow infected with parapoxvirus had severe papular and nodular lesions that completely occupied the sac and duct of the gland. The lesions were characterized by acanthosis with hyperkeratosis. Intracytoplasmic inclusion bodies were detected in the vacuolated prickle cells. By electron microscopy, mature and immature viral particles were present in the cytoplasm. These glands act as scent glands, and lesions in this organ probably affected the ecological adaptation of this individual. PMID- 9409528 TI - Hepatic carnitine palmitoyltransferase activity in cattle. AB - A fluorometric assay for the determination of hepatic carnitine palmitoyltransferase (CPT) activity was slightly modified for use with cattle samples. With this assay, the Km value was 0.56 +/- 0.10 mM with respect to L carnitine (mean +/- SD, n = 4) and was 9.6 +/- 2.2 microM (n = 3) with respect to palmitoyl-CoA. The average hepatic CPT activity was 33.6 +/- 2.0 mumol CoASH/min/g protein in 38 healthy cattle and was similar in both sexes and among breeds. Hepatic CPT activity showed no correlation with serum phospholipid, free fatty acid, triglyceride or total cholesterol concentrations. PMID- 9409529 TI - Instability of the 12-nucleotide repeat in c-myb gene of bovine T-lymphoma cells. AB - Insertion of a 12-nucleotide repeat in c-myb gene exon 9 was observed in about 15% of sporadic bovine T-lymphomas. The 12-nucleotide repeat in the T-lymphoma cells showed deletion and insertion of the repeat units during cultivation of the cells. To know whether deficiency in DNA loop repair is involved in the instability of the repeat, abilities to bind and correct the loop structure in nuclear extracts were examined. The nuclear extracts of all examined cells had ability to bind and correct the loop structure. These data suggest that instability of the 12-nucleotide repeat in bovine T-lymphoma cells might be independent of deficiency of DNA loop repair function. PMID- 9409530 TI - Inhibitory effect on LPS-induced tumor necrosis factor in calves treated with chlorpromazine or pentoxifylline. AB - The inhibitory effect of chlorpromazine (CPZ), pentoxifylline (PTX) and dexamethasone (DEX) was investigated in a model of endotoxin shock in Holstein calves following an intravenous administration of Esherichia coli endotoxin (LPS). Initial correlations with its effects on the levels of tumor necrosis factor (TNF), a pivotal mediator of endotoxin shock, and clinical signs were obtained. The pretreatment of CPZ or DEX significantly decreased the serum levels of TNF, and reduced endotoxic shock. But the pretreatment of PTX hardly reduced the increase of serum TNF levels and endotoxin shock. The levels of serum endotoxin were not significantly different a minute of postinjection of LPS in calves. The results of this study indicate that pretreatment of CPZ or DEX inhibit various biological effects on endotoxin in calves. PMID- 9409531 TI - Lymph node abscess due to Actinomyces viscosus in a cat. AB - In a four-year-old male cat, a subcutaneous phyma about 3.5 cm in diameter was surgically removed from the left inframandibular region. Histopathologically, the phyma was found to be the swollen medial retropharyngeal lymph node containing an actinomycotic abscess. The filamentous organisms in the abscess stained positively by the Gram's, Grocott's and periodic acid-Schiff methods, and were negative by the Ziehl-Neelsen method. By the immunoperoxidase method, the organisms were specifically identified as Actinomyces viscosus serotype 2 by its antiserum absorbed with A. viscosus serotype 1 antigen. PMID- 9409532 TI - Mutagenicity of dog urine determined by blue rayon extraction and the ultramicro forward-mutation method. AB - The mutagenicity of 15 household dog urine specimens were measured by the combination of blue rayon extraction and ultramicro forward-mutation method with Salmonella Typhimurium TM677 strain. A good dose-response relation was observed between the urine volume and mutation frequency. The minimum amount of urine required was 20 ml or less. The specific mutation frequency of urine greatly varied from one dog to another. The average specific mutation frequencies in the presence and absence of S9 mix were 28.7 +/- 51.5 (x 10(-4)) and 12.0 +/- 13.3 (x 10(-4)), respectively, and there was no significant difference between them. The mutation frequency markedly increased after the ingestion of broiled fish. Ten human urines specimens showed a similar level of specific mutation frequency to that of the dog urine specimens in both the presence and absence of S9 mix. PMID- 9409538 TI - Regulatory closure of cervical cytology laboratories: recommendations for a public health response. AB - The Papanicolaou test--or Pap smear test--is one of the most effective cancer screening tests available, and its ability to detect premalignant conditions has contributed to the decline in cervical cancer morbidity and mortality in the United States since its development in 1941. The success of this screening test has created confidence among women, health-care providers, and public health officials. However, this screening tool is not perfect: false-negative findings are a special concern because they can delay necessary follow-up of and treatment for women who have cervical cancer precursor lesions or invasive cervical cancer. Recent media attention has focused on cytology laboratories that have been closed as a result of deficiencies (including a high proportion of false-negative reports), and in some states legal action has been taken against individual laboratories. With the advent of revised federal regulations implementing the Clinical Laboratory Improvement Amendments (CLIA) of 1988, scrutiny of the quality of cytology laboratory practice has increased. Between 1992 and 1994, a total of 10 cytology laboratories were closed by regulatory action of the Health Care Financing Administration because they were considered a threat to the public's health. Although such closures represent <1% of CLIA-certified cytology laboratories, the attendant publicity may trigger anxiety among women. Public health officials must respond to those concerns with appropriate clinical and community actions to ensure the health and safety of women whose Pap smears were evaluated by the closed laboratories. There are no published recommendations to help develop a public health response to the regulatory closure of a cervical cytology laboratory. In April 1994, the Association of State and Territorial Public Health Laboratory Directors, through a cooperative agreement with CDC, convened a working group to provide background on the current practice of clinical cervical cytology in the United States, summarize the CLIA regulations that established specific quality assurance standards for this specialty, and recommend actions that a public health agency may initiate to deliver a measured response to laboratory closings and other regulatory sanctions. This report includes this background and summary of the workshop. The working group made three recommendations: (a) public health officials should plan for a cervical cytology laboratory closure, then, when a laboratory is closed by regulatory action, they should (b) assess the severity of the situation and determine an appropriate response and (c) provide accurate, timely information to the public. PMID- 9409533 TI - Comparison of the nuclear DNA stability against freezing-thawing and high temperature treatments between spermatozoa and somatic cells. AB - Thermostability of sperm genome against freezing-thawing and high temperature treatments was assessed by comparing the degradation patterns of genomic DNAs from epididymal sperms and somatic tissues. Golden hamster liver, kidney, epididymal sperm, and testis were frozen and thawed repeatedly, or incubated in a hot water bath. Genomic DNAs were isolated and then separated by agarose gel electrophoresis. It was revealed that the size of sperm genomic DNA was hardly changed after freezing-thawing treatment, however, the DNA sizes of the other three tissues were gradually reduced with an increasing number of freezing thawing cycles. In contrast, high temperature treatment appears to damage not only the genomic DNAs of somatic cells but also those of spermatozoa. PMID- 9409539 TI - Centenary of the birth of modern biochemistry. PMID- 9409540 TI - Ubiquitin-dependent internalization and down-regulation of plasma membrane proteins. AB - The modification of cytosolic proteins with polyubiquitin chains targets them for recognition and degradation by the multisubunit proteolytic particle, the 26S proteasome. Membrane proteins are also substrates for ubiquitination. Integral membrane proteins of the endoplasmic reticulum are ubiquitinated and destroyed by the proteasome. However, it has been shown recently that the ubiquitination of Saccharomyces cerevisiae plasma membrane proteins signals their degradation by the proteolytic system in the lysosome-like vacuole. Ubiquitination of several different classes of cell surface proteins serves as a signal for their entry into the endocytic pathway; this leads to their transport to the vacuole, where they are permanently inactivated by degradation. In yeast, ubiquitin has been implicated as an internalization signal for most, if not all, endogenous plasma membrane proteins that are known to be endocytosed. Ubiquitin-dependent internalization has been best characterized for two proteins: the mating pheromone alpha-factor receptor and the uracil permease. Some mammalian cell surface receptors are also ubiquitinated at the plasma membrane. Ubiquitination machinery is required for ligand-induced endocytosis of the growth hormone receptor, suggesting that ubiquitin-dependent endocytosis and sorting is also an important regulatory process in mammalian cells. Mammalian receptors may also be down-regulated through the degradation of their cytosolic domains by a proteasome dependent pathway. PMID- 9409541 TI - Endoplasmic reticulum degradation: reverse protein flow of no return. AB - The endoplasmic reticulum (ER) is the site of entry of proteins into the secretory pathway. It is responsible for proper folding of the proteins before delivery to their site of action. Furthermore, proofreading to detect malfolded or unnecessary proteins that have to be eliminated and regulation of protein levels are crucial ER functions. The ubiquitin-proteasome system, located in the cytoplasm, has emerged as the major ER degradation machinery. A multitude of ER resident as well as membrane-bound and soluble proteolytic substrates of the secretory pathway are retained in the ER and destined for degradation via this pathway. Their actual proteolysis is preceded by a retrograde transport to the cytoplasm. A key component of the translocation apparatus, Sec61p, is also the central subunit of the retrograde transport system. Other components of the translocon such as Sec63p or the lumenal chaperone BiP may also be involved in export to the cytosol. Novel ER membrane proteins such as Der1p, Der3p/Hrd1p, or Hrd3p might reprogram the translocon for retrograde transport. As ubiquitination is a prerequisite for degradation by the proteasome, exported proteins are ubiquitinated. Representatives of ER membrane-bound ubiquitin-conjugating enzymes, Ubc6p and Cue1p/Ubc7p, have been identified in yeast. Retrograde transport and ubiquitination seem to be coupled processes. PMID- 9409542 TI - Biogenesis of eukaryotic 20S proteasomes: the complex maturation pathway of a complex enzyme. AB - Eukaryotic 20S proteasomes harbor a remarkably complex architecture and unique proteolytic properties. Its catalytic mechanism places this enzyme in a new kind of protease family. The recently solved crystal structure of the yeast 20S complex, along with elucidation of the maturation pathway of human proteasomes, has allowed insight into structure/function relationships. Although not all of the unusual enzymatic properties such as broad substrate specificity, predominant generation of peptides with a specific size, or susceptibility to activating complexes can be explained in detail, knowledge of the structure provides important hints for an explanation of underlying mechanisms. Except for ribosome biogenesis, the complexity of eukaryotic proteasome maturation is without precedence. It is a slow process that involves a series of precisely ordered events. Proteasome structure formation is characterized by an initial cooperative formation of an alpha ring matrix, providing docking sites for a defined subset of beta subunits. Subsequent structural rearrangement allows the residual subunits to bind, followed by dimerization of two half-proteasomes. The prosequences of beta subunits exert specific functions during this process and are removed by cis- and trans-autocatalysis, most likely in the completely assembled proteasome cylinder. PMID- 9409543 TI - Regulation of ubiquitin-dependent processes by deubiquitinating enzymes. AB - An astounding number of important regulatory and structural proteins are subject to modification by the attachment of ubiquitin or ubiquitin-like proteins. This modification acts as a targeting signal, delivering the modified protein to different locations in the cell and modifying its activity, macromolecular interactions, or half-life. Deubiquitination, or the removal of this modification, is being recognized as an important regulatory strategy. This reaction is catalyzed by processing proteases known as deubiquitinating enzymes (DUBs). More than 60 DUBs are already known, although little is known about their biological roles. This review concentrates on recent findings and new insights into this fascinating class of enzymes. PMID- 9409544 TI - Pathways of ubiquitin conjugation. AB - The covalent attachment of the polypeptide ubiquitin to proteins marks them for degradation by the ubiquitin/26S proteasome-dependent degradation pathway. This pathway functions in regulating many fundamental processes required for cell viability. Phylogenetic analysis of ubiquitin sequences reveals greater variability among lower eukaryotes and defines essential residues, many of which are conserved among the three ubiquitin-like proteins known to undergo parallel ligation pathways. The hierarchical design of the ubiquitin conjugation mechanism provides great flexibility for the divergent evolution of new functions mediated by this posttranslational modification. Within this hierarchy, a single ubiquitin activating enzyme provides charged intermediates to multiple targeting pathways defined by cognate ubiquitin carrier protein (E2)/ligase (E3) pairs. Sequence analysis of E2 isozymes shows that the E2 superfamily is composed of distinct function-specific families. The apparent lack of E2/E3 specificity suggested in the literature results from the presence of multiple isozymes within many E2 families and erroneous family assignments based on incomplete data sets. Other apparent inconsistencies are explained by interfamily sequence relationships among some E2 isoforms. PMID- 9409545 TI - Mitochondria generate superoxide anion radicals and hydrogen peroxide. PMID- 9409546 TI - P-selectin is up-regulated in vital organs during murine traumatic shock. AB - Murine traumatic shock is associated with increased adherence of neutrophils to the vascular endothelium resulting in neutrophil infiltration and tissue damage. We examined the effects of trauma on the expression of the adhesion molecule P selectin in several vital organs (i.e., heart, lungs, liver, kidneys, and small intestine) 2 h after induction of Noble-Collip drum trauma in anesthetized rats. Total RNA was extracted from these organs and P-selectin mRNA was quantified by RNase protection assays. P-selectin mRNA was significantly increased over control nontraumatized, anesthetized rats in all vital organs (P<0.05 or less), with the largest increase occurring in the lung (P<0.01). Immunohistochemical analysis showed increased expression of P-selectin protein in postcapillary venules of all vital organs after trauma. To further investigate the possible mechanisms of increased P-selectin mRNA transcription promoter activity during trauma, we quantified binding of proteins from nuclear extracts to the kappaB site ( 218GGGGGTGACCC[-207]) of the P-selectin gene by electrophoretic mobility shift assay. We confirmed the results of NF-kappaB binding by demonstrating increases in p50 and p52, as well as decreases in IkappaB in cytoplasmic and nuclear extracts from the lungs of trauma rats by Western blotting. Increased activity of the transcription factor, nuclear factor kappaB (NF-kappaB), occurred in all vital organs of the trauma rats compared to sham-operated controls. Our findings suggest that severe trauma results in up-regulation of P-selectin at the transcriptional level, which is partly controlled by an NF-kappaB-responsive element in the region of the P-selectin promoter. This increased activation of NF kappB binding may contribute to the widespread increases in P-selectin expression observed in several vital organs 2 h after trauma, which in turn may play a key role in the pathogenesis of traumatic shock. PMID- 9409547 TI - Peroxynitrite modulates tyrosine-dependent signal transduction pathway of human erythrocyte band 3. AB - Peroxynitrite, the product of the reaction between nitric oxide and superoxide anion, is able to nitrate protein tyrosines. If this modification occurs on phosphotyrosine kinase substrates, it can down-regulate cell signaling. We investigated the effects of peroxynitrite on band 3-mediated signal transduction of human erythrocytes. Peroxynitrite treatment induced two different responses. At low concentrations (10-100 microM) it stimulated a metabolic response, leading to 1) a reversible inhibition of phosphotyrosine phosphatase activity, 2) a rise of tyrosine phosphorylation in the 22K cytoplasmic domain of band 3, 3) the release of glyceraldehyde 3-phosphate dehydrogenase from the membrane, and 4) the enhancement of lactate production. At high concentrations (200-1000 microM), peroxynitrite induced 1) cross-linking of membrane proteins, 2) inhibition of band 3 tyrosine phosphorylation, 3) nitration of tyrosines in the 22K cytoplasmic domain of band 3, 4) binding of hemoglobin to the membrane, 5) irreversible inhibition of phosphotyrosine kinase activity, 6) massive methemoglobin production, and 7) irreversible inhibition of lactate production. Our results demonstrate that at concentrations that could conceivably be achieved in vivo (10 100 microM), peroxynitrite behaves like other oxidants, i.e., it stimulates band 3 tyrosine phosphorylation and increases glucose metabolism. Thus, one plausible physiologic effect of peroxynitrite is the up-regulation of signaling through the reversible inhibition of phosphotyrosine phosphatase activity. At high concentrations of peroxynitrite, the tyrosine phosphorylation ceases in parallel with the nitration of band 3 tyrosines, but at these concentrations phosphotyrosine kinase activity and glycolysis are also irreversibly inhibited. Thus, at least in red blood cells, the postulated down-regulation of signaling by peroxynitrite cannot merely be ascribed to the nitration of tyrosine kinase targets. PMID- 9409548 TI - Classically conditioned changes in plasma cortisol levels induced by dexamethasone in healthy men. AB - Animal research has indicated that the activity of the hypothalamic-pituitary adrenal axis can be influenced by classical (Pavlovian) conditioning procedures. To test the hypothesis that alterations in plasma cortisol levels can be conditioned in humans, the present study used a prospective, randomized, double blind, placebo-controlled design in which a distinctively flavored beverage was paired with p.o. administration of dexamethasone. Twenty-five healthy men were randomly assigned to one of two groups. During the conditioning phase of the study, subjects in the experimental group received three conditioning trials (pairings of a distinctively flavored beverage with a capsule containing 5 mg dexamethasone) separated by 1 wk recovery periods. Subjects in the control group were treated identically, except that the capsule contained a placebo. During the test phase, all subjects underwent a test day (reexposure to the distinctively flavored beverage before receiving a placebo capsule) and a comparison day (no exposure to the beverage or the capsule). Plasma cortisol was assessed repetitively before and after administration of the beverage and capsule, as were possible confounding factors, including: behavioral variables, psychological distress, aversive reactions to the beverage, and expectations of treatment. After reexposure to the beverage and administration of a placebo capsule (conditioned stimuli), the experimental group had significantly higher levels of plasma cortisol than the control group, after controlling for variability in baseline levels of cortisol (F(5,60)=3.09; P=0.015) that could not be explained by differences in other study variables. No differences in cortisol levels were found on the comparison day. These results support the study hypothesis that changes in plasma cortisol levels can be classically conditioned in humans by pairing a distinctive beverage with p.o. administration of dexamethasone. PMID- 9409549 TI - Androgen-dependent expression of prolactin in rat prostate epithelium in vivo and in organ culture. AB - Peptide hormones and growth factors are involved in the regulation of prostatic cell proliferation, differentiation, and programmed cell death, which functions are primarily controlled by androgen. In carcinogenesis, prostatic cancer cells often lose androgen dependence and become largely dependent on local growth factors. The prostatic cancer cells able to respond to factors other than androgen by proliferation and inhibition of apoptosis are possibly able to survive. We demonstrate that prostatic epithelium expresses prolactin mRNA and protein in a characteristic manner. By using in situ hybridization, an overall distribution of prolactin mRNA was demonstrated in the epithelium of rat dorsal and lateral prostate, whereas a very specific localization of prolactin protein to single cells was observed by immunohistochemistry in the same tissues. In these cells, immunoelectron microscopy showed that prolactin was primarily localized to the secretory granules. These data demonstrate a selective regulation of prostatic prolactin at least at the level of transcript processing/translation and/or protein accumulation and secretion. In addition, the expression of prolactin protein in rat dorsal and lateral prostate was found to be androgen dependent in vivo in castrated and in castrated, testosterone treated rats, as well as in vitro in organ cultures. Our results support the concept of an autocrine/paracrine loop of prolactin action in prostate where it could mediate some of androgen actions. Also, locally synthesized prolactin might belong to the factors that take over androgen regulation of prostatic cancer cells during the development of androgen-independent growth. PMID- 9409550 TI - Melatonin maintains glutathione homeostasis in kainic acid-exposed rat brain tissues. AB - Reduced glutathione (GSH) is a key component of the cellular defense cascade against injury caused by reactive oxygen species. Because kainic acid (KA) neurotoxicity is probably mediated at least in part by oxidative stress, we examined the influence of KA treatment on GSH content and GSH-related enzyme activities in adult rats. A single injection of KA (10 mg/kg i.p.) time dependently decreased forebrain GSH (maximal reduction at 48 h). KA also markedly lowered GSH levels in amygdala and hippocampus, but not in the corpus striatum, which is resistant to KA injury. The pineal secretory product melatonin has been shown to exert neuroprotective effects against KA-induced excitotoxicity in rats. Melatonin (2.5 mg/kg i.p., administered four times) partially prevented all decreases in GSH of KA-treated rats. These neuroprotective effects of melatonin may result from a sparing of glutathione reductase, which decreased in KA-treated but not in KA/melatonin-treated animals. Moreover, KA caused a rapid decrease in the GSH content of cultured cerebellar granule neurons but not astrocytes. These cell types both express functional KA receptors, but only the former are sensitive to reactive oxygen species-dependent KA injury. Melatonin counteracted the changes in GSH induced by KA in cultured cerebellar granule neurons. Our results suggest that melatonin prevents the neurotoxic effects of reactive oxygen species linked to KA receptor activation by maintaining cellular GSH homeostasis. PMID- 9409551 TI - Induction of mitochondrial manganese superoxide dismutase in macrophages by oxidized LDL: its relevance in atherosclerosis of humans and heritable hyperlipidemic rabbits. AB - The objective of the study was to analyze the intracellular antioxidative response of macrophages (Mphi) exposed to increased levels of low density lipoprotein (LDL). We studied manganese superoxide dismutase (MnSOD) and, in part, GSH in cultured human and rabbit Mphi, and in atheromatous arterial tissue of humans and heritable hyperlipidemic (HHL) rabbits. Incubation of human Mphi with oxidized-LDL (ox-LDL) resulted in an induction of MnSOD mRNA production as shown by RT-PCR. MnSOD immunoreactivity (IR) was found to be located in the mitochondria of Mphi. In HHL rabbits, MnSOD activity and GSH concentration were significantly increased in atherosclerotic intima compared to the media of the aorta, but significantly decreased (P<0.01) in larger plaques compared with smaller ones, resulting in a significant inverse correlation of MnSOD activity (r=-0.67, P<0.001) and GSH concentration (r=-0.57, P<0.01) with plaque size. Immunohistology of the atherosclerotic intima revealed MnSOD-IR in Mac-1 (CD 11b/CD 18)-immunoreactive (ir) Mphi of human arteries and, similarly, in RAM-11 ir Mphi of rabbit ones. The relation of MnSOD-ir Mphi decreased with plaque advancement, which is consistent with biochemical findings. Most MnSOD-ir Mphi in atherosclerotic plaques revealed TUNEL-positive nuclei, indicating DNA strand breaks, and p53-IR. We conclude that mitochondrial antioxidants such as MnSOD are induced in Mphi in vitro and in atherosclerotic arteries as a reply to increased mitochondrial oxidation. As normal consequences of an increased oxidative stress due to the exposure to ox-LDL nuclear DNA strand breaks occur, which are suggested to be a signal to increase p53 protein levels. Reactive oxygen species mediated mitochondrial-dependent pathways are suggested as major contributing pathomechanisms to nuclear damage, which eventually may result in apoptosis. A common response to increased oxidative stress due to modified LDL is presumed in rabbit and human atherosclerotic plaques. PMID- 9409552 TI - Asbestos, the mesothelial cell and malignancy: a matter of life or death. PMID- 9409553 TI - Analysis of cell cycle disruptions in cultures of rat pleural mesothelial cells exposed to asbestos fibers. AB - The control of DNA integrity in mammalian cells is important to maintain the cell homeostasis and prevent neoplastic transformation. Control of cell division and cell death permits repair or elimination of damaged cells. Since asbestos fibers can produce DNA damage, chromosome alterations and apoptosis in several sorts of cells, including mesothelial cells, it was interesting to investigate cell cycle disturbances in rat pleural mesothelial cells (RPMC) treated with asbestos fibers. Cell cycle analyses were performed in RPMC exposed to crocidolite (10 and 20 microg/cm2) and chrysotile (5 and 10 microg/cm2) for different times (4 to 48 h). Both fiber types entailed a G2/M accumulation in agreement with a delay in the mitosis course. Chrysotile fibers produced a G0/G1 accumulation associated with a time-dependent p53 and p21 expression. Crocidolite exposure resulted in a delay in the G1/S transition paralleling a low rate of p53 expression. These results are in agreement with a DNA damaging potential of asbestos fibers since similar results were found following RPMC exposure to gamma rays. In asbestos treated RPMC, a low rate of apoptosis was found suggesting that RPMC may follow a DNA repair pathway that could contribute to the formation of DNA lesions. In addition, the cell cycle disturbances at the G2/M checkpoint suggest that genetically altered cells have progressed through the cycle and support the already published findings on the ability of asbestos fibers to impair cell division. PMID- 9409554 TI - Primary cell culture of human type II pneumonocytes: maintenance of a differentiated phenotype and transfection with recombinant adenoviruses. AB - Studies of the regulation of surfactant lipoprotein metabolism and secretion and surfactant protein gene expression have been hampered by the lack of a cell culture system in which the phenotypic properties of type II cells are maintained. We have developed a primary culture system that facilitates the maintenance of a number of morphologic and biochemical properties of type II pneumonocytes for up to 2 wk. Cells were isolated by collagenase digestion of midgestation human fetal lung tissue that had been maintained in organ culture in the presence of dibutyryl cyclic AMP (Bt2cAMP) for 5 days. The isolated cells were enriched for epithelial components by treatment with DEAE-dextran, plated on an extracellular matrix (ECM) derived from Madin-Darby canine kidney (MDCK) cells, and incubated at an air/liquid interface in a minimal amount of culture medium containing Bt2cAMP. The cell cultures were comprised of islands of round epithelial-like cells containing numerous dense osmiophilic granules, surrounded by sparse spindle-shaped cells with the appearance of fibroblasts. Ultrastructural examination revealed that the osmiophilic granules had the appearance of lamellar bodies, the distinguishing feature of type II pneumonocytes. Additionally, the cultures maintained elevated levels of SP-A gene expression for up to 2 wk. The expression of mRNAs encoding SP-A, SP-B, and SP-C were regulated in the cultured cells by glucocorticoids and cyclic AMP in a manner similar to that observed in fetal lung tissue in organ culture. The differentiated phenotype was most apparent when the cells were cultured at an air/liquid interface. In order to utilize the cultured type II cells for study of the effects of overexpression of various proteins and for promoter analysis, it is of essence to transfect DNA constructs into these cells with high efficiency. Unfortunately, we found the cells to be refractory to efficient transfer of DNA using conventional methods (i.e., lipofection, electroporation, or calcium phosphate-mediated transfection). However, replication-defective recombinant human adenoviruses were found to provide a highly efficient means of introducing DNA into the type II pneumonocytes. Furthermore, we observed in type II cell enriched cultures infected with recombinant adenoviruses containing the lacZ gene under control of a cytomegalovirus promoter, that beta-galactosidase was expressed uniformly in the islands of type II cells and surrounding fibroblasts. By contrast, in cultures infected with recombinant adenoviruses containing the human growth hormone (hGH) gene under control of the SP-A gene promoter and 5' flanking region, hGH was expressed only in the type II cells. Thus, this culture system provides an excellent means for identifying genomic elements that mediate type II cell-specific gene expression. PMID- 9409555 TI - Eotaxin mRNA and protein expression in chronic sinusitis and allergen-induced nasal responses in seasonal allergic rhinitis. AB - Eotaxin is an eosinophil-specific chemokine associated with the recruitment of eosinophils to the site of allergic inflammation. The aims of this study were to determine the expression of eotaxin in nasal biopsies from allergic and nonallergic individuals with chronic severe sinusitis, and to examine whether the expression of this chemokine is upregulated following allergen challenge in the nasal mucosa of patients with allergic rhinitis. We also undertook to phenotype of inflammatory cells within the submucosa expressing eotaxin mRNA. Nasal turbinate tissue from 16 individuals with allergic or nonallergic chronic sinusitis and 10 normal controls were examined for the presence of eotaxin mRNA and immunoreactivity by in situ hybridization and immunocytochemistry. The numbers of cells expressing eotaxin mRNA were also determined after either allergen or diluent challenge in atopic subjects with a history of allergic rhinitis. There was a constitutive expression of eotaxin-immunoreactivity and the presence of eotaxin mRNA-positive cells in nasal biopsies from normal individuals. Compared with normal controls, the numbers of cells expressing eotaxin mRNA and protein were significantly increased in both allergic and nonallergic sinusitis (P < 0.001). Eotaxin mRNA was expressed by nasal epithelial cells and primarily colocalized to CD68-positive macrophages within the subepithelium. In subjects with allergic rhinitis, allergen challenge markedly increased the numbers of cells expressing eotaxin mRNA and immunoreactivity in the epithelial and subepithelial cell layers (P < 0.05). This could be largely attributed to a local increase in eotaxin production within the nasal tissues. The results of this study demonstrate the constitutive expression of eotaxin and show that the numbers of cells expressing eotaxin mRNA are increased within the epithelial and subepithelial layers of the nasal mucosa in individuals with chronic sinusitis. Furthermore, allergen challenge of the nasal mucosa in atopic subjects results in a local upregulation of eotaxin expression. These data suggest a potential role for this chemokine in the pathogenesis of allergic and nonallergic eosinophilic inflammation characterizing chronic sinusitis and allergic rhinitis. PMID- 9409556 TI - Increased type I procollagen mRNA in airways and pulmonary vessels after vagal denervation in rats. AB - To test the hypothesis that increased airway strain resulting from lung denervation initiates a fibroproliferative response in the airways, we compared the transcriptional expressions of alpha1(I)-procollagen and tropoelastin in the lungs of rats subjected to unilateral vagal denervation, unilateral vagal denervation combined with ipsilateral phrenectomy, or thoracotomy without denervation (controls). We found increases in alpha1(I)-procollagen messenger ribonucleic acids (mRNAs) in the submucosa of the airways and the adventitia of airways and pulmonary vessels of the denervated lungs in 31% of the rats subjected to unilateral denervation (with and without phrenectomy), and in none of the controls. The increased transcripts were associated with collagen deposition in the peribronchial and perivascular tissue, and occasionally with cell proliferation leading to occlusion of the airway and vascular lumina. Unilateral phrenectomy did not decrease the frequency with which production of Type I procollagen was upregulated, suggesting that the upregulation was not entirely dependent on airway strain. Tropoelastin expression was not influenced by denervation. Our results indicate that the autonomic nervous system has a previously unsuspected trophic influence on collagen synthesis in the airways and pulmonary vessels. Abolition of this influence by denervation may lead to structural changes analogous to those observed in bronchiolitis obliterans after lung transplantation. PMID- 9409557 TI - Kinetics and quantitation of eosinophil and neutrophil recruitment to allergic lung inflammation in a brown Norway rat model. AB - We quantitated neutrophil and eosinophil migration into lung parenchyma using specific peroxidase enzyme assays, and into the bronchoalveolar compartment by bronchoalveolar lavage (BALF), in sensitized brown Norway (BN), Fischer, and Lewis rats and also assessed the lungs by histopathology. Fourteen days after sensitization with ovalbumin (OA in alum [given subcutaneously] and OA with Bordetella pertussis [given intraperitoneally]), rats were challenged with an OA aerosol for 1 h. In BN rats, there was marked perivascular and peribronchial edema, focal hemorrhages, and increase in lung wet weight and BALF protein content, accompanied by neutrophilic infiltration at 3-14 h postchallenge. Few eosinophils were seen at 14 h in lung tissue or in BALF. Neutrophils peaked at 24 h in parenchyma ([94 +/- 7] x 10[6]) and in BALF ([2.7 +/- 0.4] x 10[6]) and declined rapidly thereafter. Marked eosinophil infiltration into parenchyma was apparent by 24 h. Eosinophil accumulation peaked at 48 h in parenchyma ([127 +/- 18] x 10[6]) and at 72 h in BALF ([10 +/- 2.4] x 10[6]), comprising up to 85% of lavage cells at this time. Lung eosinophilia persisted for at least 6 d with only a slow decline or clearance, not approximating baseline until day 13 after challenge. Histopathology showed peribronchial and interstitial eosinophilic pneumonia, most severe on day 3. In contrast to the BN rats, essentially no pulmonary inflammation was observed in Lewis and Fischer rats. This model in the BN rat, and the specific peroxidase assays for quantitating tissue eosinophils and neutrophils, should be useful for investigating the regulation of allergen induced eosinophil and neutrophil migration into and clearance from the lung. PMID- 9409559 TI - Cytokine-induced neutrophil transepithelial migration is dependent upon epithelial orientation. AB - The mechanisms by which mediators and cytokines stimulate neutrophils to migrate across the lung epithelium are still unclear. We hypothesized that neutrophil transepithelial migration depends upon polarity of the epithelium. We therefore compared neutrophil migration through human lung Type II-like alveolar epithelial cell line (A549) monolayers grown on the upper versus lower surface of permeable filters to simulate apical-to-basal and basal-to-apical movement of neutrophils, respectively. The classic chemoattractants formyl-methionylleucylphenylalanine (FMLP), leukotriene B4 (LTB4), and interleukin-8 (IL-8) induced equivalent neutrophil transepithelial migration in the apical-to-basal and basal-to-apical directions. However, the degree of neutrophil transepithelial migration was significantly greater in the basal-to-apical direction in response to either IL 1beta or tumor necrosis factor-alpha (TNF-alpha). Enhanced TNF-alpha-induced neutrophil migration through A549 monolayers in the basal-to-apical direction occurred regardless of whether the TNF-alpha was above or below the filter/monolayer complex. Actinomycin D pretreatment of A549 monolayers had no effect on FMLP-induced neutrophil transepithelial migration, but markedly (about 75%) inhibited both TNF-alpha- and IL-1beta-induced neutrophil transepithelial migration, regardless of monolayer orientation. Thus, in contrast to classic chemoattractants, IL-1beta and TNF-alpha induced greater neutrophil transepithelial migration in a basal-to-apical direction, and this occurred independently of the cytokine location, but depended upon intact metabolic capacity of the A549 cells. These data suggest that the mechanisms important for neutrophil transepithelial migration in response to classic chemoattractants differ from those important for migration in response to inflammatory cytokines. PMID- 9409558 TI - Elevation of manganese superoxide dismutase gene expression by thioredoxin. AB - Manganese superoxide dismutase (MnSOD) is a mitochondrial enzyme that dismutates potentially toxic superoxide radical into hydrogen peroxide and dioxygen. This enzyme is critical for protection against cellular injury due to elevated partial pressures of oxygen. Thioredoxin (TRX) is a potent protein disulfide reductase found in most organisms that participates in many thiol-dependent cellular reductive processes and plays an important role in antioxidant defense, signal transduction, and regulation of cell growth and proliferation. Here we describe induction of manganese superoxide dismutase by thioredoxin. MnSOD mRNA and activity were increased dramatically by low concentrations of TRX (28 microM). Elevation of MnSOD mRNA by TRX was inhibited by actinomycin D, but not cycloheximide, occurring both in cell lines and primary human lung microvascular endothelial cells. mRNAs for other antioxidant enzymes including copper-zinc superoxide dismutase and catalase were not elevated, demonstrating specificity of induction of MnSOD by TRX. Thiol oxidation by diamide or alkylation by chlorodinitrobenzene inhibited MnSOD induction, further indicating a requirement for reduced TRX. Because both oxidized and reduced thioredoxin (28 microM) induced MnSOD mRNA, the intracellular redox status of externally added Escherichia coli oxidized TRX was determined. About 45% of internalized E. coli TRX was reduced, with 8% in fully reduced form and about 37% in partially reduced form. However, when TRX reductase and nicotinamide adenine dinucleotide (NADPH) were added to the extracellular medium with TRX, more than 80% of E. coli TRX was found to be in a fully reduced state in human adenocarcinoma (A549) cells. Although lower concentrations of oxidized TRX (7 microM) did not induce MnSOD mRNA, this concentration of TRX, when reduced by NADPH and TRX reductase, increased MnSOD mRNA six-fold. In additional studies, MCF-7 cells stably transfected with the human TRX gene had elevated expression of MnSOD mRNA relative to vector-transfected controls. Thus, both endogenously produced and exogenously added TRX elevate MnSOD gene expression. These findings suggest a novel mechanism involving reduced TRX in regulation of MnSOD. PMID- 9409560 TI - IL-12 and IFN-gamma are required for initiating the protective Th1 response to pulmonary cryptococcosis in resistant C.B-17 mice. AB - A murine model was used to assess the role of cytokines in initiating protective T-cell-mediated immunity in the lung. A pulmonary infection was initiated by intratracheal inoculation of Cryptococcus neoformans (Cne). Previously, we had established that Cne lung clearance was mouse-strain-specific: C.B-17 mice were resistant and developed a Th1-like response, whereas C57BL/6 mice were susceptible and did not develop a Th1 response. In the present study we showed that monoclonal anti-interferon-gamma (IFN-gamma) and anti-interleukin-12 (IL-12) antibody administration prior to infection of resistant C.B-17 mice inhibited lung clearance of Cne. Cytokine profiles of lung and lung-associated lymph nodes (LALN) from monoclonal antibody (mAb)-treated C.B-17 mice were switched from Th1 like to Th2-like, and mAb-treated C.B-17 mice exhibited lung eosinophilia, which was absent in control C.B-17 mice. Additionally, C.B-17 mice treated with anti IFN-gamma and anti-IL-12 mAb demonstrated a significantly lower percentage of lung macrophages expressing inducible nitric oxide synthase (iNOS) than did control mice. These studies clearly demonstrate that both IFN-gamma and IL-12 are required for initiation of a Th1 response in resistant C.B-17 mice. PMID- 9409561 TI - Dietary restriction mitigates ozone-induced lung inflammation in rats: a role for endogenous antioxidants. AB - Studies were undertaken to determine whether dietary restriction protects against acute pulmonary oxidant challenge. Male F344 rats were fed NIH-31 diet either ad libitum or at restricted levels equal to 75% that of ad libitum intake. After 3 wk of dietary adaptation, animals were exposed by inhalation to 2.0 ppm ozone (O3) for 2 h or chamber air and evaluated for cellular and biochemical indices of pulmonary toxicity. Compared to air controls, bronchoalveolar lavage fluid (BALF) from O3 exposed ad libitum fed rats contained increased protein (145 versus 380 microg/ml), PMN infiltration (0 versus 11%) and fibronectin (45 versus 607 U/ml). Diet restriction abrogated these indicators of pulmonary inflammation induced by ozone. Binding of 18O3 to BALF protein and cells was significantly decreased in diet restricted rats while BALF ascorbate and glutathione levels, but not alpha tocopherol or urate, were elevated compared to ad libitum fed rats. Taken together, these results indicate that dietary restriction affords protection against O3-induced oxidant toxicity. Protection is mediated partially by increases in ascorbate in the fluid bathing the lung surface, thereby providing an antioxidant sink which minimizes the ability of O3 to reach biological targets. PMID- 9409562 TI - Prostaglandins induce vascular endothelial growth factor in a human monocytic cell line and rat lungs via cAMP. AB - Prostaglandins have emerged as a therapeutic option for patients with peripheral vascular disease as well as pulmonary hypertension as a means to increase blood flow. We tested the hypothesis that prostaglandins regulate vascular endothelial growth factor (VEGF) expression in the human monocytic THP-1 cell line and in isolated perfused rat lungs. Our data show that the stable PGI2-analogue iloprost induces VEGF gene expression (predominantly VEGF121, but also VEGF165 isoforms) and VEGF protein synthesis in THP-1 cells. This effect is abolished by dexamethasone and by Rp-cAMP, a specific inhibitor of cAMP-dependent protein kinase (PKA) activation. The calcium channel blocker diltiazem has no effect on the iloprost-induced VEGF gene expression, and depletion of intracellular Ca2+ stores by long-term exposure (16 h) of THP-1 cells to thapsigargin does not inhibit iloprost-induced VEGF gene expression, suggesting that an increase in intracellular Ca2+ is not essential for VEGF gene induction by iloprost. However, an increase of intracellular Ca2+ by a short-term (2 h) exposure of THP-1 cells to thapsigargin or to the calcium-ionophore A23187 increases VEGF mRNA levels, indicating that a change in intracellular Ca2+ by itself can alter VEGF gene expression. The effects of thapsigargin or A23187 on VEGF gene expression are also mediated via cAMP-PKA since they are inhibited by Rp-cAMP. In isolated perfused rat lungs, PGI2 and PGE2 increases VEGF mRNA abundance whereas Rp-cAMP inhibits the prostaglandin-induced VEGF gene activation. Thus, our data suggest that prostaglandins stimulate VEGF gene expression in monocytic cells and in rat lungs via a cAMP-dependent mechanism. PMID- 9409563 TI - Roles of adhesion molecules ICAM-1 and alpha4 integrin in antigen-induced changes in microvascular permeability associated with lung inflammation in sensitized brown Norway rats. AB - Increased microvascular permeability and mucosal edema are pathological features of airway inflammation in asthma. In this study, we investigated the characteristics of the edema response occurring in a model of antigen-induced lung inflammation in sensitized brown Norway rats and examined the effects of monoclonal antibodies (mAbs) to adhesion molecules on this response. Ovalbumin (OA) challenge-induced increases in lung permeability were determined by the leakage of 125I-labeled bovine serum albumin (BSA) into the extravascular tissues of the lungs 24 h after challenge in animals intravenously injected (prechallenge) with this tracer. Inflammatory cell infiltration into the alveolar space was determined by bronchoalveolar lavage (BAL). Mean extravascular plasma volume in the lung increased 233% as compared with control (P < 0.005) at 24 h and increased to 517% by 72 h. The 24-h edema response was completely inhibited by two oral doses (0.1 mg/kg) of dexamethasone 1 h before, and 7 h after, challenge. Intraperitoneal administration of the anti-rat ICAM-1 mAb 1A29, or anti-rat alpha4 integrin mAb TA-2 (2 mg/kg at 12 and 1 h before, and 7 h after, antigen challenge), significantly suppressed eosinophil infiltration into the alveolar space without inhibiting the enhanced microvascular leakage and lung edema. Determination of plasma antibody concentrations by ELISA of mouse IgG1 indicated that sufficient concentrations of the appropriate mAb were present to block alpha4- or ICAM-1-dependent adhesion. The results suggest that increases in microvascular permeability and plasma leakage occurred independently of eosinophil accumulation. PMID- 9409564 TI - Role of IFN-gamma in the inhibition of the allergic airway inflammation caused by IL-12. AB - T-helper 2 (Th2)-like cells are thought to play a crucial role in the pathogenesis of the eosinophilic airway inflammation observed in asthma. In a murine model of allergen-induced airway eosinophilia and bronchial hyperresponsiveness (BHR), we have shown that interleukin (IL)-12 can suppress antigen-induced airway changes despite the presence of circulating specific IgE. In the present study, we investigated the role of interferon-gamma (IFN-gamma) in the inhibitory effects of IL-12 on allergic airway inflammation. Repeated daily exposure of actively immunized mice to aerosolized ovalbumin (OVA), as compared with aerosolized saline (SAL), induced a significant increase in bronchoalveolar lavage fluid (BALF) eosinophilia and OVA-specific serum IgE in both IFN-gamma receptor-deficient (IFN-gammaR KO) and wild-type mice. As compared with placebo (PLAC), administration of recombinant murine IL-12 (rmIL-12) during the daily aerosol exposure (but not at the time of immunization) significantly inhibited BALF eosinophilia in both IFN-gammaR KO mice and wild-type controls, without influencing the production of specific IgE. In contrast, administration of rmIL 12 during the active immunization inhibited both BALF eosinophilia and specific IgE in wild-type mice as compared with littermates given PLAC; however, treatment with rmIL-12 during immunization, in comparison with PLAC, caused a significant increase in BALF eosinophilia and specific IgE in IFN-gammaR KO mice. These results demonstrate that inhibition of the allergen-induced eosinophil influx in murine airways by IL-12 is IFN-gamma-dependent during the initial sensitization, but becomes IFN-gamma-independent during the secondary response. PMID- 9409565 TI - Presidential address to the European Surgical Association: research and surgery. PMID- 9409566 TI - Cost-effectiveness of extraperitoneal laparoscopic inguinal hernia repair: a randomized comparison with conventional herniorrhaphy. Coala trial group. AB - OBJECTIVE: To determine the cost-effectiveness of laparoscopic inguinal hernia repair. SUMMARY BACKGROUND DATA: Laparoscopic inguinal hernia repair seems superior to open techniques with respect to short-term results. An issue yet to be studied in depth remains the cost-effectiveness of the procedure. As part of a multicenter randomized study in which >1000 patients were included, a cost effectiveness analysis from a societal point of view was performed. METHODS: After informed consent, all resource costs, both in and outside the hospital, for patients between August 1994 and July 1995 were recorded prospectively. Actual costs were calculated in a standardized fashion according to international guidelines. The main measures used for the evaluation of inguinal hernia repair were the number of averted recurrences and quality of life measured with the Short Form 36 questionnaire. RESULTS: Resource costs were recorded for 273 patients, 139 in the open and 134 in the laparoscopic group. Both groups were comparable at baseline. Average total hospital costs were Dfl 1384.91 (standard deviation: Dfl 440.15) for the open repair group and Dfl 2417.24 (standard deviation: Dfl 577.10) for laparoscopic repair, including a disposable kit of Dfl 676. Societal costs, including costs for days of sick leave, were lower for the laparoscopic repair and offset the hospital costs by Dfl 780.83 (75.6%), leaving the laparoscopic repair Dfl 251.50 more expensive (Dfl 4665 versus Dfl 4916.50). At present, the recurrence rate is 2.6% lower after laparoscopic repair. Thus, 38 laparoscopic repairs, costing an additional Dfl 9,557, prevent the occurrence of one recurrent hernia. Quality of life was better after laparoscopic repair. CONCLUSION: A better quality of life in the recovery period and the possibility of replacing parts of the disposable kit with reusable instruments may result in the laparoscopic repair becoming dominantly better--that is, less expensive and more effective from a societal perspective. PMID- 9409567 TI - Prospective, randomized trial on the effect of cyclic versus continuous enteral nutrition on postoperative gastric function after pylorus-preserving pancreatoduodenectomy. AB - OBJECTIVE: The effect of a cyclic versus a continuous enteral feeding protocol on postoperative delayed gastric emptying, start of normal diet, and hospital stay was assessed in patients undergoing pylorus-preserving pancreatoduodenectomy (PPPD). SUMMARY BACKGROUND DATA: Delayed gastric emptying occurs in approximately 30% of patients after PPPD and causes prolonged hospital stay. Enteral nutrition through a catheter jejunostomy is used to provide postoperative nutritional support. Enteral infusion of fats and proteins activates neurohumoral feedback mechanisms and therefore can potentially impair gastric emptying and prolong postoperative gastroparesis. METHODS: From September 1995 to December 1996, 72 consecutive patients underwent PPPD at the Academic Medical Center, Amsterdam. Fifty-seven patients were included and randomized for either continuous (CON) jejunal nutrition (0-24 hr; 1500 kCal/24 hr) or cyclic (CYC) enteral nutrition (6 24 hr; 1125 kCal/18 hr). Both groups had an equal caloric load of 1 kCal/min. The following parameters were assessed: days of nasogastric intubation, days of enteral nutrition, days until normal diet was tolerated orally, and hospital stay. On postoperative day 10, plasma cholecystokinin (CCK) levels were measured during both feeding protocols. RESULTS: Nasogastric intubation was 9.1 days in the CON group (n = 30) and 6.7 days in the CYC group (n = 27) (not statistically significant). First day of normal diet was earlier for the CYC group (15.7 vs. 12.2 days, p < 0.05). Hospital stay was shorter in the CYC group (21.4 vs. 17.5 days, p < 0.05). CCK levels were lower in CYC patients, before and after feeding, compared with CON patients (p < 0.05). CONCLUSIONS: Cyclic enteral feeding after PPPD is associated with a shorter period of enteral nutrition, a faster return to a normal diet, and a shorter hospital stay. Continuously high CCK levels could be a cause of prolonged time until normal diet is tolerated in patients on continuous enteral nutrition. Cyclic enteral nutrition is therefore the feeding regimen of choice in patients after PPPD. PMID- 9409568 TI - Influence of preoperative transarterial lipiodol chemoembolization on resection and transplantation for hepatocellular carcinoma in patients with cirrhosis. AB - OBJECTIVE: To investigate the impact of preoperative transarterial lipiodol chemoembolization (TACE) in the management of patients undergoing liver resection or liver transplantation for hepatocellular carcinoma. PATIENTS AND METHODS: TACE was performed before surgery in 49 of 76 patients undergoing resection and in 54 of 111 patients undergoing liver transplantation. Results were retrospectively analyzed with regard to the response to treatment, the type of procedure performed, the incidence of complications, the incidence and pattern of recurrence, and survival. RESULTS: In liver resection, downstaging of the tumor by TACE (21 of 49 patients [42%]) and total necrosis (24 of 49 patients [50%]) were associated with a better disease-free survival than either no response to TACE or no TACE (downstaging, 29% vs. 10% and 11 % at 5 years, p = 0.08 and 0.10; necrosis, 22% vs. 13% and 11% at 5 years, p = 0.1 and 0.3). Five patients (10%) with previously unresectable tumors could be resected after downstaging. In liver transplantation, downstaging of tumors >3 cm (19 of 35 patients [54%]) and total necrosis (15 of 54 patients [28%]) were associated with better disease-free survival than either incomplete response to TACE or no TACE (downstaging, 71 % vs. 29% and 49% at 5 years, p = 0.01 and 0.09; necrosis, 87% vs. 47% and 60% at 5 years, p = 0.03 and 0.14). Multivariate analysis of the factors associated with response to TACE showed that downstaging occurred more frequently for tumors >5 cm. CONCLUSIONS: Downstaging or total necrosis of the tumor induced by TACE occurred in 62% of the cases and was associated with improved disease-free survival both after liver resection and transplantation. In liver resection, TACE was also useful to improve the resectability of primarily unresectable tumors. In liver transplantation, downstaging in patients with tumors >3 cm was associated with survival similar to that in patients with less extensive disease. PMID- 9409569 TI - Prospective evaluation of Pringle maneuver in hepatectomy for liver tumors by a randomized study. AB - OBJECTIVE: To evaluate whether vascular inflow occlusion by the Pringle maneuver during hepatectomy can be safe and effective in reducing blood loss. SUMMARY BACKGROUND DATA: Hepatectomy can be performed with a low mortality rate, but massive hemorrhage during surgery remains a potentially lethal problem. The Pringle maneuver is traditionally used during hepatectomy to reduce blood loss, but there is a potential harmful effect on the metabolic function of hepatocytes. There has been no prospective randomized study to determine whether the Pringle maneuver can decrease blood loss during hepatectomy, improve outcome, or affect the metabolism of hepatocytes. METHODS: From July 1995 to February 1997, we studied 100 consecutive patients who underwent hepatectomy for liver tumors. The patients were randomly assigned to liver transection under intermittent Pringle maneuver of 20 minutes and a 5-minute clamp-free interval (n = 50), or liver transection without the Pringle maneuver (n = 50). The surface area of liver transection was measured and blood loss during transection per square centimeter of transection area was calculated. Routine liver biochemistry, arterial ketone body ratio (AKBR), and the indocyanine green (ICG) clearance test were done. RESULTS: The two groups were comparable in terms of preoperative liver function and in the proportion of patients having major hepatectomy. The Pringle maneuver resulted in less blood loss per square centimeter of transection area (12 mL/cm2 vs. 22 mL/cm2, p = 0.0001), a shorter transection time per square centimeter of transection area (2 min/cm2 vs. 2.8 min/cm2, p = 0.016), a significantly higher AKBR in the first 2 hours after hepatectomy, lower serum bilirubin levels in the early postoperative period, and, in cirrhotic patients, higher serum transferrin levels on postoperative days 1 and 8. The complication rate, the hospital mortality rate, and the ICG retention at 15 minutes on postoperative day 8 were equal for the two groups. CONCLUSION: Performing the Pringle maneuver during liver transection resulted in less blood loss and better preservation of liver function in the early postoperative period. This is probably because there was less hemodynamic disturbance induced by the bleeding. PMID- 9409570 TI - First-line chemotherapy improves the resection rate and long-term survival of locally advanced (T4, any N, M0) squamous cell carcinoma of the thoracic esophagus: final report on 163 consecutive patients with 5-year follow-up. AB - OBJECTIVE: The objective of this prospective, nonrandomized study was to evaluate the immediate and long-term results of first-line chemotherapy and possible surgery in locally advanced, presumably T4 squamous cell esophageal cancer. SUMMARY BACKGROUND DATA: Locally advanced esophageal cancer is rarely operable and has a dismal prognosis. For this reason, neoadjuvant cytoreductive treatments are more and more frequently used with the aim of downstaging the tumor, increasing the resection rate, and possibly improving survival. METHODS: From January 1983 to December 1991, 163 consecutive patients with a presumedly T4 squamous cell carcinoma of the thoracic esophagus (group A) received on average 2.5 cycles (range, 1-6) of first-line chemotherapy with cisplatin (100 mg/m2 on day 1) and 5-fluorouracil (1000 mg/m2 per day, in continuous infusion from day 1 through day 5). Chemotherapy was followed by surgery when adequate downstaging of the tumor was obtained. RESULTS: Chemotherapy toxicity was WHO grade 0 to 2 in 80% of cases, but 3 toxic deaths (1.9%) occurred. Restaging suggested a downstaging of the tumor in 101 of 163 patients (62%), but only 85 patients (52%) underwent resection surgery; it was complete or R0 in 52 (32%) and incomplete or R1-2 in 33. Overall postoperative mortality was 11.7% (10 of 85), morbidity 41% (35 of 85). Complete pathologic response was documented in 6 patients, and significant downstaging to pStage I, IIA, or IIB occurred in 25 more patients. The overall 5-year survival was 11 % (median, 11 months). After resection surgery, the 5-year survival was 20% (median, 16 months); none of the nonresponders survived 4 years after palliative treatments without resection (median survival, 5 months). The 5-year survival rate of the 52 patients undergoing an R0 resection was 29% (median, 23 months). Stratifying patients according to the R, pT, pN, and pStage classifications, the survival curves were comparable to the corresponding data obtained in the 587 group B patients with "potentially resectable" esophageal cancer who underwent surgery alone during the same period. Furthermore, the results were improved in comparison with 136 previous or subsequent patients with a locally advanced tumor who did not undergo neoadjuvant treatments (group C). In these patients, the R0 resection rate was 7%, and the overall 5-year survival was 3% (median, 5 months). CONCLUSION: Although nonrandomized, these results suggest that in locally advanced esophageal carcinoma, first-line chemotherapy increases the resection rate and improves the overall long-term survival. In responding patients who undergo R0 resection surgery, the prognosis depends on the final pathologic stage and not on the initial pretreatment stage. PMID- 9409571 TI - Is Barrett's esophagus the precursor of most adenocarcinomas of the esophagus and cardia? A biochemical study. AB - OBJECTIVE: To obtain biochemical evidence that Barrett's esophagus (BE) is the precursor of most adenocarcinomas (Adc) of the esophagus and cardia. SUMMARY BACKGROUND DATA: Based on morphologic data, BE was previously proposed as the precursor of most Adc of the esophagus. This hypothesis would receive strong support if biochemical evidence were found to demonstrate a pattern common to BE and Adc of the esophagus and cardia. METHODS: We studied the presence of intestinal-type proteins sucrase-isomaltase (SI) and crypt Cell Antigen (CCAg) in BE, Barrett's Adc, and esophageal-cardial Adc without BE. In each case specimens were collected from normal esophagus, stomach, tumor, and BE mucosa when present. To study related conditions, five specimens of peptic esophagitis and of squamous cell carcinoma were also analyzed. An indirect immunofluorescence technique was employed and sections were analyzed with laser confocal microscopy imaging. RESULTS: Most Barrett's mucosa specimens stained positively for SI (93%) and CCAg (89%). These proteins were detected in BE independently of the type of metaplasia, the coexistence of dysplasia, or the presence of associated Adc. SI and CCAg were present in 25 (96%) and 24 (92%) of the cases of Adc respectively. No statistical difference was detected in SI and CCAg expression between Adc samples with and without BE, between BE and Adc samples with or without BE, and between tumors located in the esophagus versus the cardia. No staining for these proteins was detected in stomach or esophageal mucosa, in submucosal glands of the esophagus, in peptic esophagitis or squamous cell carcinoma. CONCLUSION: These data show that BE and Adc of the esophagus and cardia have a similar phenotype and support the hypothesis that most of these tumors probably originate from preexisting BE. PMID- 9409572 TI - Clinical value of extended biologic staging by bone marrow micrometastases and tumor-associated proteases in gastric cancer. AB - OBJECTIVE: To investigate whether extended staging, including biologic grading and aspects of an early systemic disease component, would give additional prognostic information on patients with curatively resected gastric cancer. BACKGROUND: Tumor-associated proteolytic mechanisms have been shown to be essential for invasion and metastasis. The urokinase-type plasminogen activator (uPA) system is of major biologic impact, but different interactive proteases and inhibitors with modulating effects also must be considered. The detection of early tumor cell dissemination indicates the systemic character of a primarily local gastric cancer. The confrontation of the organism with these cells determines the often unpredictable course of an individual tumor after presumed curative primary treatment. METHODS: In a prospective study of 247 consecutive patients with gastric cancer, detection of disseminated tumor cells in bone marrow aspirates was immunocytochemically performed in 180 patients. The expression of uPA, activators of uPA (cathepsin D, antithrombin III), uPA substrates (plasminogen, matrix-metalloproteinase 2 [collagenase IV, 72 kD; MMP 2]), uPA/plasmin inhibitors (plasminogen activator inhibitor type 1 and 2 [PAI-1, PAI-2], alpha1-antitrypsin, alpha2-antiplasmin), uPA receptor (uPA-R), and parameters of the uPA-R cycle (alpha2-macroglobulin, alpha1-antichymotrypsin) could be determined immunohistochemically and were scored semiquantitatively in 203 patients. Kaplan-Meier statistical techniques and multivariate Cox regression models were used for prognostic analyses. RESULTS: In multivariate analysis considering all the established risk factors, disease-free survival was independently predicted by PAI-1 (relative risk 2.21, 1.32-3.73) and cathepsin D (relative risk 2.98, 1.28-6.91) besides pT, pN, and extended resection. Tumor cell dissemination was found to be an additional prognostic factor in early tumor stages (pT1, T2) and lymphnode-negative patients. Stepwise regression analysis revealed an extended staging system with new risk groups. Node-positive, curatively resected pT1/2 patients with low expression of PAI-1 had a favorable prognosis (mean recurrence-free survival [MRT] 54.84 months), similar to that of node-negative patients (MRT 54.76 months). In node-negative, curatively resected pT1/2 patients, detection of bone marrow tumor cells and high expression of PAI-1 defined a subgroup with a steep decrease of prognosis (MRT 36.60 months), which was worse than that of node-positive patients (MRT 45.81). CONCLUSION: This new staging model gives better prognostic differentiation of subgroups, which should be considered in future adjuvant therapy protocols. In addition, it indicates that the uPA system might be a future therapeutic target. PMID- 9409573 TI - Ileocecal segment transposition does not alter whole gut transit in humans. AB - OBJECTIVES: We have recently described a reservoir for rectal replacement after total mesorectal excision for rectal carcinoma. The ileocecal segment with its intact extrinsic nerve and blood supply is placed between the ascending colon and the anal canal. This reconstruction has been shown to provide good defecation quality and anorectal function. Whether gastric emptying and small as well as large bowel transit are affected by this transposition remains unclear. Our aim was to quantify whole gut transit in such patients and compare it with that of a matched group of controls. METHODS: Gastric emptying rates and small intestinal and colonic transit times were assessed scintigraphically in 12 patients aged 46 to 87 years with ileocecal reservoir reconstruction after total mesorectal excision and compared to a sex-matched group of asymptomatic healthy volunteers of similar age. Gastric emptying rates and small intestinal and colonic transit times were calculated as described previously. Data were compared using Wilcoxon's signed rank test for gastric emptying rates and small bowel transit or by analysis of variance for colonic transit; p < 0.05 was considered significant. RESULTS: Gastric time for half of the meal (T50) was 161 +/- 16 minutes for patients and 201 +/- 22 for the controls. Small bowel transit time was 150 +/- 15 minutes for patients and 177 +/- 22 for the controls. Geometric center at 6 hours was 1.53 +/- 0.13 for patients and 1.27 +/- 0.16 for the controls. Geometric center at 24 hours was 2.96 +/- 0.23 for patients and 2.57 +/- 0.25 for the controls. Data are mean +/- SEM. SUMMARY: Gastric emptying rates and small bowel transit and colonic transit times (expressed as geometric center at 6 and 24 hours) were similar in patients with ileocecal reservoir reconstruction and in a sex- and age-matched group of healthy controls. We conclude that the transposition of an ileocecal segment with intact extrinsic neurovascular supply between the sigmoid colon and the anal canal does not alter whole gut transit, not even in any of the presumably key regions. PMID- 9409575 TI - The surgical personality: fact or fiction. PMID- 9409576 TI - Thomas G. Orr Memorial Lectureship. Colon cancer from etiology to prevention. AB - BACKGROUND: Recent changes in healthcare have resulted in a general malaise among physicians leading to negative advice to young people about career choices in the medical field. The author uses his experiences in academic surgery to show that medicine continues to be an exciting and desirable career choice. DATA SOURCES: Relevant literature from surgery, toxicology, molecular biology, carcinogenesis, and epidemiology fields. CONCLUSIONS: Significant advances have occurred in the understanding of colon cancer. These advances have occurred because of extensive collaborations across many different scientific fields. The methods used and the collaborations involved provide a model for other physicians to unravel the mysteries of disease and to make academic medicine fun and exciting. The risk of colon cancer depends on genetically determined factors that combine with environmental exposure (diet) to determine risk. Microwaving meat before cooking and eating more cruciferous vegetables may reduce this risk. Additional prevention steps are in the planning stages. PMID- 9409577 TI - Edgar J. Poth Memorial/W.L. Gore and Associates, Inc. Lectureship. The clinical and pathological spectrum of recurrent carotid stenosis. AB - BACKGROUND: Hemodynamically significant (> or =50%) carotid restenosis occurs in approximately 10% to 12% of individuals undergoing carotid endarterectomy. The underlying pathology is usually neointimal thickening within 3 years and recurrent atherosclerosis thereafter. Although a number of etiologic factors have been implicated in the development of restenosis, the etiology remains unclear and preventative measures relatively ineffective. METHODS: A review of the English literature was undertaken to determine the incidence, clinical presentation, and pathologic features of carotid restenosis. CONCLUSIONS: Carotid restenosis is the major factor limiting long-term patency after carotid endarterectomy. Although drug therapy has been shown to be effective in preventing restenosis in animal models, the results of clinical human trials have been disappointing. Delineation of the biochemical and molecular mechanisms contributing to the development of restenosis is essential if effective therapeutic interventions are to be developed. PMID- 9409574 TI - Intrauterine tracheal obstruction, a new treatment for congenital diaphragmatic hernia, decreases amniotic fluid sodium and chloride concentrations in the fetal lamb. AB - OBJECTIVE: To evaluate the effect of fetal tracheal occlusion on sodium and chloride concentrations in amniotic and tracheal fluid. SUMMARY BACKGROUND DATA: Intrauterine tracheal occlusion has been proposed to reverse pulmonary hypoplasia, an important prognostic factor in congenital diaphragmatic hernia. In early human trials, technical failure of the obstructive device has been reported. METHODS: Eight fetal lambs (gestational age = 95 days) were subjected to fetal tracheoscopy, and amniotic and tracheal fluid samples were taken. In multiple pregnancies (n = 6), amniotic fluid was also sampled from the contralateral amniotic sac and used as a control. Subsequently, endotracheal obstruction, using a detachable balloon, was performed. After 14 days, all fetuses were delivered, and sodium and chloride concentrations in amniotic and tracheal fluid were measured again. Statistical analysis was done using a two tailed Student's t test, paired or unpaired as appropriate. RESULTS: In controls, between 95 and 109 days gestational age, no significant changes occurred in sodium or chloride concentrations in amniotic or tracheal fluid. After 2 weeks of tracheal obstruction, however, chloride and sodium concentrations in amniotic fluid decreased (chloride = 76.7 mEq/L vs. 107.6 mEq/L, p = 0.0003; sodium = 109.6 mEq/L vs. 125.9 +/- 5.2 mEq/L, p = 0.019). A concomitant increase in chloride and sodium concentration was observed in tracheal fluid (chloride = 145.4 mEq/L vs. 130.0 mEq/L, p = 0.047; sodium = 153.1 mEq/L vs. 142.9 mEq/L, p = 0.051). When comparing groups at 109 days, chloride and sodium concentrations in amniotic fluid were markedly lower in the treated group versus controls (p = 0.0004 and p = 0.05 for chloride and sodium, respectively). CONCLUSION: Complete tracheal occlusion in ovine fetuses results in a significant decrease of amniotic fluid sodium and chloride concentrations. PMID- 9409578 TI - Claude H. Organ, Jr. Honorary/Sandoz Nutrition Lectureship. Ethics in research and surgical practice. AB - The topic "Ethics in Research and Surgical Practice" appears most appropriate for the 1997 Claude H. Organ, Jr. Honorary Sandoz Nutrition Lecture. With the increasing numbers of ethical problems facing clinicians today, greater emphasis on ethics is demanded. This lecture focuses primarily on the clinical applications of basic science and ethics certainly plays a major role in this regard. Ethical principles espoused by the American College of Surgeons and the American Medical Association will be emphasized. Further, the results of the deliberations of the Advisory Committee on Human Radiation experiments will be discussed. Certainly, the Tuskegee experiment debacle will be mentioned. Ethics remains important if we are to give our patients the best care. We must always keep in the vanguard of our thinking that the welfare of our patients comes first. High ethical principles emphasize our obligations and responsibilities to our patients. PMID- 9409579 TI - Why use clinical pathways rather than practice guidelines? AB - BACKGROUND: Financial pressures from managed care organizations, the government, and other "stakeholders" have resulted in the production of practice guidelines and clinical pathways. Clinical pathways involve all segments of a health care system and may prove to be more beneficial and less hazardous to patients and health care providers. METHODS: A historical narrative describing the development of clinical pathways by the Southwestern Surgical Congress (SWSC) and the Southeastern Surgical Congress (SESC) is made. The motivations, the benefits, and the hazards of both clinical pathways and practice guidelines are discussed. RESULTS: Clinical pathways have proven to reduce length of stay (LOS), complications, and cost, and provide increased patient satisfaction whereas practice guidelines from some specialties show improved quality of care when compared with nonspecialists. However, many practice guidelines are developed by specialists on "best practice" standards, and few have documented studies proving their effectiveness. CONCLUSIONS: Eleven clinical pathways were developed by the SWSC and the SESC and are in the process of revision and study for efficacy. They will be disseminated in the American Surgeon and on the SWSC web site for review and comment. In 1998, both congresses hope to publish the efficacy of selected pathways by describing their effect on LOS and charge for those diagnostic related groups. PMID- 9409580 TI - Endoscopic ultrasound for peripancreatic masses. AB - BACKGROUND: Pancreatic neoplasms can be difficult to diagnose and stage preoperatively. Accurate staging allows the surgeon to select which patients can benefit from resection versus palliative therapy. Endoscopic ultrasound (EUS) with endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) is a diagnostic modality that provides visualization of peripancreatic tumors and their relationship to the surrounding structures as well as enabling cytologic diagnosis of the tumor and adjacent lymphadenopathy. METHODS: To define the role of this technique, a retrospective study was performed on 20 patients in the past year with peripancreatic tumors. RESULTS: Twelve men and 8 women ranging in age from 28 to 84 years (mean 67) were included in the study. Each patient underwent computed tomography followed by EUS-FNA, and the results were compared with operative findings or clinical course. The EUS-FNA findings included 10 pancreatic ductal carcinomas (50%), 5 pancreatitis (25%), 2 spindle cell neoplasms (10%), 1 cholangiocarcinoma (5%), 1 cystadenoma (5%), and 1 metastatic breast carcinoma (5%). Overall, EUS-FNA led to a significant change in the management of 12 patients (60%) through either diagnosing benign pathology, upstaging of the carcinoma, or determination that the peripancreatic mass represented a metastatic lesion. Five patients underwent resection of their peripancreatic tumors, and 3 patients had palliative procedures. Operative findings corresponded with EUS-FNA in all 8 patients. The 5 patients diagnosed with pancreatitis continued to be followed up for the possibility of a false negative FNA, but to date none have developed malignancy. CONCLUSIONS: EUS-FNA is a useful tool for the imaging and staging of peripancreatic tumors and will aid in the proper preoperative selection of patients who will benefit from resectional therapy. PMID- 9409581 TI - Surgical morbidity, mortality, and long-term survival in patients with peripancreatic cancer following pancreaticoduodenectomy. AB - BACKGROUND: Surgical resection of the primary tumor in peripancreatic cancer has been associated with an improved survival and decreased morbidity in the recent literature. The purpose of this review was to analyze the results at a single institution. METHODS: Between 1985 and 1995, 88 patients underwent a pancreaticoduodenectomy for adenocarcinoma of the pancreatic head region and had complete long-term follow-up. Patient records were reviewed to determine morbidity, mortality, and survival. RESULTS: Tumor histology included pancreatic head adenocarcinoma (n = 46), ampullary adenocarcinoma (n = 28), duodenal adenocarcinoma (n = 8), and cholangiocarcinoma (n = 6). Morbidity occurred in 26 patients (29%). Perioperative mortality was seen in 6 patients (7%). No perioperative mortality was seen over the last 3 years, which included 33 patients. The mean follow-up was 29 months, with a median survival of 19 months. At last follow-up, 24 patients were alive without disease with an average survival of 43 months (1 to 141). There were 54 patients who died with cancer with an average survival of 21 months (1 to 117). Based on Kaplan and Meier statistical analysis the estimated survival was 47% at 2 years and 25% at 5 years. The location of the primary tumor (P = 0.0006) and the presence of positive lymph nodes (P = 0.05) was shown to have a negative impact on survival. CONCLUSION: Pancreaticoduodenectomy can be done with acceptable morbidity and mortality. The outlook with this disease remains poor, but long-term survival can be achieved in some patients. PMID- 9409582 TI - Neoadjuvant chemoradiation for extrahepatic cholangiocarcinoma. AB - BACKGROUND: The prognosis for patients with extrahepatic bile duct cancer remains poor. The purpose of this study was to evaluate our initial results with preoperative chemoradiation for extrahepatic cholangiocarcinoma, in the context of our experience with conventional treatment of this disease over the past 13 years. METHODS: From 1983 through 1996, analysis of all patients treated for extrahepatic cholangiocarcinoma was performed. RESULTS: Of 91 total patients, 51 had unresectable disease and 40 underwent resection. Median survival was significantly different for patients who underwent resection (22.2 months) versus those treated palliatively (10.7 months; P <0.0001). Nine patients underwent preoperative chemoradiation (5 perihilar, 4 distal) prior to resection. Three patients in the preoperative chemoradiation group had a pathologic complete response, while the remainder showed varying degrees of histologic response to treatment. The rate of margin-negative resection was 100% for the preoperative chemoradiation group versus 54% for the group who did not receive preoperative chemoradiation (P <0.01). There were no major intra-abdominal complications in the patients treated with preoperative chemoradiation. CONCLUSIONS: These results suggest that preoperative chemoradiation for extrahepatic bile duct cancer can be performed safely, produces significant antitumor response, and may improve the ability to achieve tumor-free resection margins. Additional trials of preoperative chemoradiation are warranted. PMID- 9409583 TI - Cytoablative therapy with combined resection and cryosurgery for limited bilobar hepatic colorectal metastases. AB - BACKGROUND: Cryosurgery can be employed in patients with unresectable hepatic metastases when the tumor size and the number of metastases are limited. However, local recurrence can result from incomplete ablation. We proposed a trial of complete cytoablation with a combined approach of cryosurgery and hepatic resection for patients with bilobar hepatic metastases. METHODS: Seven patients underwent cryosurgery alone (CRYO). Seven additional patients underwent combined resection and cryosurgery (CRYO+RES) for bilobar metastases. RESULTS: In the CRYO group, 5 of 7 patients had at least one centrally located tumor. All 5 of these patients had early recurrence at the site of ablation. In the CRYO+RES group complete ablation was achieved in 7 of 7. Two (28.6%) of these patients developed local recurrence. CONCLUSION: Cytoablation of hepatic metastases can be safely achieved with combined hepatic resection and cryosurgery in selected patients. Long-term survival data are necessary before advocating widespread application of this approach. PMID- 9409584 TI - Cryosurgical ablation of hepatic tumors. AB - BACKGROUND: Cryosurgical ablation of hepatic tumors relies on nonspecific tissue necrosis due to freezing as well as microvascular thrombosis. Patients with selected primary and metastatic hepatic malignancies who are not candidates for surgical resection are afforded potentially curative benefit using this technique. METHODS: Forty patients underwent cryosurgery for hepatic malignancy related to colorectal metastasis (n = 27), hepatocellular carcinoma (n = 8), metastatic breast (n = 2), metastatic neuroendocrine (n = 2), and metastatic ovarian carcinoma (n = 1). Intraoperative ultrasound (IOUS) was used in all patients to help locate the tumor and guide the cryosurgical trocar to the lesions. RESULTS: Indications for cryosurgical ablation included bilobar and centrally located disease, poor medical risk, insufficient hepatic reserve, and involved margin after wedge resection. Major complications included hepatic parenchyma cracking requiring transfusion in 5 patients, 1 postoperative biliary stenosis, and 1 inferior vena cava injury. There were 3 postoperative deaths from non-hepatic-related events. Based on Kaplan-Meier analysis the estimated overall survival for patients with hepatocellular carcinoma (60% at 18 months) was compared with patients with colorectal metastases (30% at 18 months). Nine patients (23%) are currently free of disease with an average follow-up of 17.7 months. The pattern of failure was identified at the site of cryosurgical ablation in 2 of 88 lesions. CONCLUSIONS: Cryosurgical ablation of selected hepatic malignancies is a safe and viable treatment for patients not amenable to surgical resection. PMID- 9409585 TI - Gallbladder cancer and trocar site recurrences. AB - BACKGROUND: Critics of laparoscopic surgery cite an increased incidence of tumor recurrence at the trocar sites following laparoscopic cholecystectomy in patients incidentally found to have carcinoma of the gallbladder. The purpose of this review was to determine if laparoscopic cholecystectomy performed in patients with gallbladder cancer results in an increased incidence of abdominal wall recurrences. METHODS: The charts of all patients with gallbladder cancer registered at the University of Texas M. D. Anderson Cancer Center from January 1991 through April 1996 were retrospectively reviewed. Data were collected on initial and subsequent surgical procedures, tumor grade and histology, T stage, adjuvant therapy, and survival. These data were analyzed with regard to abdominal wall recurrences and outcome. RESULTS: Ninety-three patients with gallbladder cancer were seen during this period; 79 patients with complete follow-up information comprised the study population. Comparison of the incidence of abdominal wall recurrences among the categories of surgical procedure (laparoscopic versus open versus laparoscopic converted to open) did not reveal any statistically significant differences. Overall 5-year survival was 10%. CONCLUSIONS: Gallbladder cancer is an aggressive malignancy with few long-term survivors. In addition, these data show that the incidence of abdominal wall implantation is not increased with laparoscopic surgery but is more likely a manifestation of the aggressive nature of this tumor. PMID- 9409587 TI - Clinical observation of the temporal association between crack cocaine and duodenal ulcer perforation. AB - HYPOTHESIS: To determine if a cause-effect relationship exists between crack cocaine use and duodenal ulcer perforation (DUP). PATIENTS AND METHODS: A retrospective study was conducted of all patients undergoing emergency surgical management for peptic ulcer disease over a 6-year period at a large inner-city municipal teaching hospital. The hospital records of 78 consecutive patients presenting with complications of peptic ulcer disease between April 1990 and April 1996 were reviewed. Group A (n = 24) consisted of patients with confirmation of crack cocaine usage within 8 hours of clinical presentation; group B (n = 54) consisted of patients with no antecedent history of crack cocaine use. Demographic data, timing of drug use, clinical presentation, laboratory and radiographic findings, toxicology screening, operative findings, and postoperative course were compared between the two groups. RESULTS: Both groups revealed a similar gender distribution, tobacco use, prior peptic ulcer symptoms, and laboratory findings. Group A patients were younger (t test, P = 0.01) and more likely to present with perforation, whereas patients in group B presented with a combination of symptoms (chi square, P = 0.03). Duodenal ulcer perforation was present in 75% of patients in group A compared with 46% of patients in group B (chi square, P = 0.04). Group B patients had a significantly longer hospital stay compared with those in group A (t test, P = 0.01). Both crack cocaine and alcohol are independent predictors of duodenal ulcer perforation. CONCLUSIONS: Patients with recent use of crack cocaine and/or alcohol are more likely to present with duodenal perforations. Although a temporal association between crack cocaine use and duodenal ulcer perforation was demonstrated, this study does not confirm a cause-effect relationship. A prospective cohort study is needed to clarify the pathogenesis of this potential cause-effect relationship. PMID- 9409586 TI - Percutaneous endoscopic gastrostomy tube placement in a surgical training program. AB - BACKGROUND: This study examines the patterns of use of percutaneous endoscopic gastrostomy (PEG) and primary open gastrostomy (Gtube) performed in a residency training program in surgery. METHODS: A retrospective cohort study that assesses the indications and outcomes of 317 PEGs and 75 isolated Gtubes used for gastric access between 1987 and 1997. RESULTS: The demographics and risk factors of the patients receiving Gtube and PEG were comparable. The mean number of PEGs performed per resident is currently 13 per year (mean 5 over 10 years) with a 97% PEG success rate; an 88% success rate is demonstrated for placement of jejunal extensions. CONCLUSIONS: PEGs are generally preferable to Gtubes as primary procedures. Surgical residents should become competent in PEG placement by performing adequate numbers of procedures with fully trained staff. PMID- 9409588 TI - Esophageal manometry and 24-hour pH testing in the management of gastroesophageal reflux patients. AB - BACKGROUND: With rising interest in gastroesophageal reflux disease, an evaluation of the importance of manometry (M) and 24-hour pH testing (pH) for decisions regarding these patients is appropriate. METHODS: Two gastroenterologists and two surgeons were presented with history and physical examination, endoscopy, histology, and esophagram data ("DATA") from 100 patients and asked to make a treatment decision. After some time, either pH or M was added to DATA, and a further decision requested. Finally, DATA plus pH plus M was presented, and a decision was requested. Decisions were evaluated for changes in medical therapy, changes between medical and surgical therapy, and changes in type of surgery offered. RESULTS: Overall, 43% (173 of 400) of decisions were altered by the addition of both M and pH to DATA, with 28.5% (114 of 400) of decisions changed from medical therapy to surgery or vice versa by the addition of both tests to DATA. The addition of M alone changed decisions more often than pH alone especially with regard to the type of surgery offered (P <0.05). CONCLUSIONS: Together, M and pH alter clinical decisions and often alter the decision regarding surgery. Both tests appear important, but M more frequently alters overall management decisions and the type of surgery offered. Despite the need for cost containment, these clinical tools are essential to important decisions regarding the care of patients with gastroesophageal reflux disease. PMID- 9409589 TI - Respiratory symptoms in patients with gastroesophageal reflux disease following medical therapy and following antireflux surgery. AB - BACKGROUND: It is not known whether antireflux surgery is more effective than medical therapy to control respiratory symptoms (RS) in gastroesophageal reflux disease (GERD). METHODS: In 21 GERD patients with RS, reflux was assessed by endoscopy, manometry, and pH monitoring. Patients had proton pump inhibitor therapy and cisapride for 6 months. After GERD relapsed following withdrawal of medical therapy, 7 patients with normal esophageal peristalsis had a laparoscopic Nissen fundoplication and 14 with impaired peristalsis a Toupet fundoplication. Respiratory symptoms were scored prior to treatment, at 6 months following medical therapy, and at 6 months after surgery. RESULTS: Heartburn and esophagitis were effectively treated by medical and surgical therapy. Only surgery improved regurgitation. Respiratory symptoms improved in 18 patients (85.7%) following surgery and in only 3 patients (14.3%) following medical therapy (P <0.05). Esophageal peristalsis improved following the Toupet fundoplication. CONCLUSION: Medical therapy fails to control reflux since it does not inhibit regurgitation. Surgery controls reflux and improves esophageal peristalsis, which contributes to its superiority over medical therapy in the treatment of RS associated with GERD. PMID- 9409590 TI - Free tissue transfer to extend the limits of limb salvage for lower extremity tissue loss. AB - BACKGROUND: The extent of tissue loss amenable to primary healing after revascularization is unknown. Salvage of limbs with large soft-tissue defects with exposed tendon, joint, or bone lies beyond the limits of conventional techniques. We report our results using free tissue transfer as an adjunct to lower extremity vascular reconstruction in patients with complex ischemic or infected wounds. METHODS: Retrospective chart review of patient and wound characteristics. RESULTS: From January 1992 to June 1996, 585 procedures were performed in 544 patients, including 27 free flaps in 26 patients: 17 free flaps combined with distal bypass (7 staged, 10 simultaneous) and 10 isolated free flaps. Flap donor sites included radial forearm (8), latissimus dorsi (7), rectus abdominus (9), and scapula (3). Surgical indications included extensive ischemic/neurotrophic ulcers, and nonhealing vein graft harvest incision or transmetatarsal amputation site. Mean area of tissue loss was 70 cm2, mean ulcer duration was 5 months, and 92% of patients had exposed tendon, joint, or bone. During a mean follow-up of 14 months, 2 patients died of cardiopulmonary disease and 3 flaps failed, resulting in below-knee amputation. Six flaps were revised for graft stenosis (1), venous thrombosis (1), or flap edge necrosis (4). Limb salvage rate was 70% at 24 months by life-table analysis. Functional ambulation was achieved in 21 of 24 (88%) patients, including 7 of 8 with diabetes, end stage renal disease, and heel ulcers. CONCLUSION: In select ambulatory patients with large soft-tissue defects and exposed deep structures, functional limb salvage is obtainable in more than 80% of patients. For lesions not amenable to vascular reconstruction with conventional methods of wound coverage, free tissue transfer extends the limits of limb salvage and is a viable alternative to amputation. PMID- 9409591 TI - Cost savings associated with the nonroutine use of carotid angiography. AB - BACKGROUND: To evaluate the economic impact of performing carotid endarterectomy based only on a diagnosis of duplex scanning, we evaluated a cohort of patients treated at our institution during 1 calendar year. METHODS: Ninety-seven patients were evaluated and divided into two groups: those with and without arteriogram prior to their operation. Duplex scan and arteriogram results were reviewed to determine their effect on the operative plan. Hospital charges and physician fees were assessed for each patient admission. Operative results, complications, and total charges were compared between the two groups. RESULTS: There was one operative stroke in each group for a stroke rate of 2%. Angiographic complications included one stroke and one femoral artery thrombosis. Two arteriograms led to a change in the operative plan. The hospital charges for patients without an arteriogram was $10,292 verses $13,906 for patients with an arteriogram (P < 0.01). Physician charges for patients without an arteriogram were $3,882, with angiograms and $6,297. The total charges related to the endarterectomy were $14,174 and $20,203, respectively. Arteriograms accounted for an increase of 43% in total charges. CONCLUSION: Nonroutine use of angiography does not increase operative risk or postoperative length of stay, and preoperative angiography increases total charges by 43% ($6,029) per patient. PMID- 9409592 TI - A statewide, hospital-based analysis of frequency and outcomes in carotid endarterectomy. AB - BACKGROUND: For more than 40 years carotid endarterectomy (CE) has been used in the treatment of extracranial carotid disease for the prevention of stroke. Recent prospective clinical trials have confirmed the benefit of CE for both symptomatic and asymptomatic patients. Our purpose was to examine statewide trends in the numbers of CE over a 6-year time period and to evaluate outcomes. METHODS: Using data from the North Carolina Medical Database Commission (NCMDC) all CE procedures from 1988 to 1993 were identified. Numbers of CE were compared with the population and hospital admissions. Variables of length of stay, hospital charges, discharge disposition, and occurrence of stroke and death were analyzed. RESULTS: A total of 11,973 CE were performed in 6 years. Compared by admissions, population, and the proportion of elderly, the number of CE increased yearly. The stroke rate was 1.7% and the death rate 1.2% for an overall in hospital stroke plus mortality rate of only 2.7%. CONCLUSIONS: From a diverse group of hospitals and a large number of surgeons and patients, this hospital based study documents the acceptance and safety of CE in the treatment of extracranial carotid disease. PMID- 9409593 TI - Descending thoracic aorta-to-femoral artery bypass grafts. AB - BACKGROUND: Descending thoracic aorta-to-femoral artery (DTAFA) bypass graft is an alternative procedure to revascularize lower limbs. METHODS: Between 1976 and 1996, 41 patients underwent DTAFA bypass grafts. Operative indications consisted of previous abdominal graft thrombosis (22 cases, group 1), abdominal operations (8, group 2), initial vascular operation in the presence of difficult aortas (6, group 3), and infection of aortic grafts (5, group 4). RESULTS: Perioperative mortality was 5%. Cumulative 10-year primary patency, limb salvage, and survival rates were 64%, 79%, and 55%, respectively. Breaking down the result on the basis of the four groups, DTAFA bypass grafts performed for infection of previous aortic grafts had a significantly lower primary patency rate (25% at 24 months; P < 0.004) with dismal limb salvage (31% at 24 months; P < 0.001) and survival rates (0% at 24 months; P < 0.005). CONCLUSIONS: DTAFA bypass grafts can be safely and durably used in patients who had thrombosis of previous abdominal grafts or had a difficult abdomen or as the initial vascular operation in the presence of difficult aortas. Conversely, dismal results are obtained in the treatment of aortic graft infection. PMID- 9409594 TI - Prospective characterization and selective management of the abdominal compartment syndrome. AB - BACKGROUND: The abdominal compartment syndrome (ACS) is now recognized as a frequent confounder of surgical critical care following major trauma; however, few prospective data exist concerning its characterization, evolution, and response to decompression. METHODS: Acutely injured patients with an injury severity scale (ISS) score >15 requiring emergent laparotomy and intensive care unit (ICU) admission were prospectively evaluated for the development of ACS. The syndrome was defined as an intra-abdominal pressure (IAP) >20 mm Hg complicated by one of the following: peak airway pressure (PAP) >40 cm H2O, oxygen delivery index (DO2I) <600 mL O2/min/m2, or urine output (UO) <0.5 mL/kg/hr. Physiologic response to decompression was similarly documented prospectively. RESULTS: Over a 14-month period ending December 1995, 21 (14%) of 145 patients (ISS >15) requiring laparotomy and admitted to our surgical ICU developed ACS; mean age was 39 +/- 9 years; injury mechanism was blunt in 60%; ISS 26 +/- 6. At initial laparotomy, 67% underwent abdominal packing (57% for major liver injuries). Mean IAP was 27 +/- 2.3 mm Hg, and time from laparotomy to decompression was 27 +/- 4 hours; 24% were planned whereas the remaining were prompted by deteriorating organ function as defined above (cardiopulmonary in 43%; renal in 19%; both renal and cardiopulmonary in 14%). Following decompression, there was an increase in cardiac index, oxygen delivery, urine output, and static compliance while there was a decrease in pulmonary capillary wedge pressure, systemic vascular resistance, and peak airway pressure. CONCLUSIONS: The abdominal compartment syndrome occurs in a significant number of severely injured patients, and it develops quickly (27 +/- 4 hours). Cardiopulmonary deterioration is the most frequent reason prompting decompression. Timely decompression of the ACS results in improvements in cardiopulmonary and renal function. These data support the use of the proposed ACS grading system for selective management of the syndrome. PMID- 9409595 TI - Trends in the management of hepatic injury. AB - BACKGROUND: Options for management of blunt hepatic injury have broadened to include both operative management (OM) and nonoperative management (NOM). We identify trends in evaluation and management of blunt hepatic injury at a level 1 trauma center. METHODS: Charts of 106 patients with blunt hepatic injuries from July 1, 1991 to June 30, 1995 were reviewed for method of abdominal evaluation (computed tomography versus DPL), injury severity score, liver injury grade, method of management, length of stay (LOS), transfusion requirements, complications, and outcome. RESULTS: Nonoperative management steadily increased to 86%. Successful NOM occurred in 96% (48 of 50) and was not related to injury grade. Transfusion requirements were significantly greater in the group with OM versus those with NOM (11.3 versus 2.7). Patients with NOM also had significantly shorter intensive care unit stay and total LOS. CONCLUSIONS: The majority of patients with blunt liver injury can be successfully managed nonoperatively regardless of injury grade. Nonoperative management may allow decreased resource utilization because of shorter hospital stays and decreased transfusion requirements. PMID- 9409596 TI - Selective management of penetrating neck trauma based on cervical level of injury. AB - BACKGROUND: Selective surgical exploration of penetrating neck wounds is now the standard of care, but the safety and cost effectiveness of selective diagnostic testing remains controversial. We herein review our 18-year prospective evaluation of a progressively selective approach. PATIENTS AND METHODS: Since 1979, 312 patients sustained penetrating trauma to the anterior neck; 75% were stabbed and 24% were shot. Zone I was penetrated in 13%, zone II in 67%, and zone III in 20%. RESULTS: In all, 105 (34%) of the patients had early exploration (16% were nontherapeutic). Of the 207 (66%) observed, 1 (0.5%) required delayed exploration. Length of stay was 8.0 days following exploration, 5.1 days following negative exploration, and 1.5 days following observation. In the last 6 years, 40% have had adjunctive testing: 69% of zone I, 15% of zone II, and 50% of zone III injuries. CONCLUSION: Selective management of penetrating neck injuries is safe and does not mandate routine diagnostic testing for asymptomatic patients with injuries in zones II and III. PMID- 9409597 TI - Is operating room resuscitation a way to save time? AB - BACKGROUND: Direct admission to the operating room (OR) can shorten the time to incision. A protocol for operating room resuscitation was established with patient triage based on (1) cardiac arrest, (2) hypotension unresponsive to field fluid resuscitation, or (3) uncontrolled external hemorrhage. METHODS: Operating room resuscitation over 11 years was reviewed to determine whether the triage criteria correctly identified patients requiring operation. Survival was analyzed and compared with the probability of survival (Ps) determined at the scene. RESULTS: Operating room resuscitation patients were more likely to require a major operation regardless of mechanism of injury. Of 476 patients with penetrating injury, 170 patients had persistent low blood pressure (<90 mm Hg), and 146 (85.9%) of these required major operative intervention. The mean time to incision in this group was 21.7-67.5 minutes less than for patients not receiving OR resuscitation. Observed survival was significantly greater than that predicted for this group. CONCLUSIONS: Field triage criteria are able to reliably identify patients who require immediate major operative intervention. Direct admission to the OR results in a more timely initiation of operative therapy for patients requiring emergency surgical procedures. PMID- 9409598 TI - Early aggressive treatment for Merkel cell carcinoma improves outcome. AB - BACKGROUND: Merkel cell carcinoma (MCC) is a rare and aggressive neuroendocrine tumor of dermal origin. Treatment recommendations are limited owing to a paucity of retrospective data and an absence of prospective data. The objective of this study was to determine current therapeutic trends and their impact upon outcome. METHODS: A retrospective study (1983 to 1996) was performed with patients from the Department of Defense and our University-affiliated hospitals. RESULTS: Thirty-five patients were evaluated with a mean follow-up of 31 months. Overall, 1- and 2-year survival rates were 80% and 50%, respectively. Patients undergoing wide local excision, prophylactic lymph node dissection, and adjuvant radiotherapy had significantly decreased locoregional and distant recurrence rates and improved survival when compared with their counterparts. Adjuvant chemotherapy did not diminish recurrence rates nor improve survival. Both locoregional and distant recurrence significantly decreased survival. CONCLUSIONS: These data suggest that early aggressive treatment for MCC improves both tumor control and survival, whereas the early use of chemotherapy does not improve outcome. PMID- 9409599 TI - Use of fine needle aspiration for solid breast lesions is accurate and cost effective. AB - BACKGROUND: Palpable breast tumors have traditionally been diagnosed with open biopsy or core biopsy. We propose fine needle aspiration biopsy (FNA) as a reliable, cost-saving initial procedure in these patients. METHODS: Eighty-five palpable solid breast masses of the breast in 85 patients were classified by a combination of physical examination, mammography, and/or ultrasound as probably benign, indeterminate, or highly suspicious for cancer. All tumors had FNA biopsies. All patients had either a confirmatory open biopsy (55) or close clinical follow-up (30) with a mean follow-up of 29 months (range 6 to 36). RESULTS: Thirty-four patients classified as clinically benign had a benign FNA biopsy. No cancers were detected in this group by either open surgical biopsy or clinical follow-up. Twenty patients were classified clinically as indeterminate. All had FNA biopsies, and 6 were either positive for cancer or suspicious for cancer. Fourteen patients had negative FNA biopsies. Five of the 6 abnormal biopsies had cancer on open biopsies. The 1 false-positive result occurred in a lactating patient. Thirty-one patients were classified clinically as highly suspicious for cancer. Twenty-three were confirmed as cancer with FNA biopsy. Eight needed open surgical biopsy to confirm cancer. All 31 patients clinically suspicious for cancer had cancer. In patients classified clinically as highly suspicious or probably benign, FNA was a reliable first diagnostic step (100% positive predictive value, 100% specificity, 87% sensitivity, and 89% negative predictive value). CONCLUSIONS: Fine needle aspiration biopsy of solid palpable breast lesions should be the diagnostic procedure of choice for those patients classified clinically as probably benign or clinically as highly suspicious for cancer. Cost analysis revealed elimination of an open biopsy in such cases would save $1,100 per patient. For highly suspicious cases, a negative fine needle aspiration should not deter an open surgical biopsy. For patients classified as indeterminate, fine needle aspiration biopsy results are not reliable enough to determine treatment. PMID- 9409600 TI - Stereotactic and ultrasound core needle breast biopsy performed by surgeons. AB - BACKGROUND: The authors evaluated outcomes and treatment costs of stereotactic core needle biopsy (SCNB) and ultrasound core needle biopsy (UCNB), and needle localization biopsy (NLB) in managing patients with mammographic abnormalities presenting to the surgeon. METHODS: Data for all patients with mammographic lesions who underwent SCNB or UCNB since their introduction at this institution were prospectively collected over 17 months. Mean inclusive costs of the three procedures were accumulated and compared. RESULTS: Stereotactic core needle biopsy was performed for 342 lesions in 319 women, for a malignancy rate of 19%; UCNB was performed for 157 lesions in 144 patients, yielding a malignancy rate of 17%. With a mean follow-up of 13.5 months, 1 patient with in situ carcinoma was diagnosed late. Absolute cost savings for the period studied was $721,963. CONCLUSIONS: Minimally invasive breast biopsy procedures can safely and reliably be performed by surgeons in clinical practice with increased patient convenience and decreased costs. PMID- 9409601 TI - Preoperative 5-fluorouracil and radiation therapy for locally advanced breast cancer. AB - BACKGROUND: Fifteen percent of breast cancer patients present with large tumors involving skin or chest wall. Often, surgery with primary wound closure is impossible. We used neoadjuvant chemoradiation in locally advanced breast cancer patients, in hopes of increasing resectability. METHODS: Eligible patients had locally advanced breast cancer deemed unresectable with primary wound closure. Patients received 8 weeks of infusional 5-fluorouracil (5-FU) 200 mg/m2 per day and radiation therapy to 50 Gy. Patients rendered resectable underwent modified radical mastectomy (MRM) followed up by chemotherapy. RESULTS: Of 30 evaluable patients, 73% had an objective clinical response. All were able to undergo MRM with primary wound closure; 63% had residual disease, 20% had minimal microscopic disease, and 17% had complete pathologic response. Treatment-related toxicity was minimal. Surgical morbidity was not increased. CONCLUSIONS: Infusional 5-FU with concomitant radiotherapy is well tolerated and effective at producing shrinkage in the majority of patients, converting inoperable breast cancer to easily resectable disease. PMID- 9409602 TI - Successful treatment of esophageal achalasia with laparoscopic Heller myotomy and Toupet fundoplication. AB - BACKGROUND: Recently, investigators have reported the use of endoscopic myotomy in the treatment of esophageal achalasia. As with the open operation, considerable disagreement exists regarding the appropriate length of the myotomy and the need for a concomitant antireflux procedure. METHODS: Patients presenting with symptomatic achalasia between 1993 and 1997 were included in this prospective study. Preoperative studies included barium upper gastrointestinal study, endoscopy, and esophageal manometry. Laparoscopic myotomy was completed in all 20 patients; 18 had concomitant Toupet fundoplication. RESULTS: Operative times ranged from 95 to 345 minutes (mean 216). Blood loss ranged from 50 to 300 cc (mean 100 cc). There were 7 minor complications (5 mucosal injuries repaired laparoscopically, 1 bile leak and 1 splenic capsular tear). Nine patients began a liquid diet on the first day postoperatively; 19 were tolerating liquids by postoperative day 3. Hospital stay ranged from 2 to 20 days (mean 5). Eighteen patients had complete relief of dysphagia, with less than one reflux episode per month. One individual continues to have mild persistent solid food dysphagia. Another patient initially did well but subsequently developed mild recurrent dysphagia and reflux. One patient required laparoscopic take-down of the wrap because of recurrent dysphagia and now has no problems swallowing, but does complain of mild reflux. Two other patients also have mild reflux, 1 of whom did not undergo fundoplication. CONCLUSIONS: Laparoscopic Heller myotomy can be performed safely with excellent results in patients with achalasia. Adding a partial fundoplication appears to help control postoperative symptoms of reflux. This procedure should be considered the procedure of choice in patients with symptomatic esophageal achalasia. PMID- 9409603 TI - Laparoscopic Nissen fundoplication for gastroesophageal reflux disease. AB - BACKGROUND: This is a retrospective review from March 1994 through October 1996 of 78 patients who underwent laparoscopic Nissen fundoplication for gastroesophageal reflux disease (GERD). The purpose of this study was to evaluate the postoperative results and complications. METHODS: The patient profile included 38 men and 40 women with a mean age of 46 years (range 11 to 81). The main preoperative symptoms included heartburn (92%), respiratory problems (42%), and dysphagia (32%). Preoperative assessment included esophagogastroduodenoscopy, upper gastrointestinal series, esophageal manometry, and 24-hour pH monitoring. Essential indications for surgery included esophagitis (83%), Barrett's esophagus without dysplasia (22%), and esophageal stricture (23%). All patients underwent a 360-degree wrap with a Maloney dilator and division of the short gastric vessels. RESULTS: The mean operative time was 206 minutes (range 90 to 455). The average time for patients to tolerate a full liquid diet was 1.2 days, and the mean hospital stay was 2.4 days. Current follow-up, from 3 to 36 months, showed complete resolution of heartburn without medications in 67 patients (86%), occasional heartburn in 8 patients (10%), and slight improvement of heartburn in 3 patients (4%). Five patients with preoperative Barrett's metaplasia showed no evidence of it postoperatively (n = 2) or marked regression (n = 3). CONCLUSION: Laparoscopic Nissen fundoplication is the procedure of choice for patients with complicated GERD. PMID- 9409604 TI - Comparison of preoperative endoscopic retrograde cholangiopancreatography and laparoscopic cholecystectomy with operative management of gallstone pancreatitis. AB - BACKGROUND: Preoperative endoscopic retrograde cholangiopancreatography (ERCP) with laparoscopic cholecystectomy (ERCP/LC) should reduce hospital stay when compared with laparoscopic cholecystectomy/intraoperative cholangiogram (LC/IOC) and selective common bile duct exploration (CBDE). METHOD: Retrospective review of 82 patients with gallstone pancreatitis. RESULTS: Thirty-one patients had preoperative ERCP/LC and 51 patients underwent LC/IOC. Nineteen percent in the ERCP/LC group developed postprocedure pancreatitis. The presence of choledocholithiasis was associated with an increased incidence of post-ERCP pancreatitis (38%) and an increased length of stay ([LOS] 24 versus 10 days). The development of post-ERCP pancreatitis markedly increased LOS to 30 days. Six percent in the LC/IOC group developed postoperative pancreatitis. The mean LOS was 10.1 days. Open CBDE increased the LOS to 14.5 days. Postoperative pancreatitis increased the LOS to 23 days. The LOS of patients with choledocholithiasis who underwent uncomplicated ERCP/ES or LC/IOC with open CBDE was similar. CONCLUSIONS: The development of postprocedure pancreatitis is a more important determinant of hospital stay than an open operative procedure. LC/IOC with its lower incidence of postprocedure pancreatitis resulted in a shorter hospital LOS even when open CBDE was performed. PMID- 9409605 TI - High negative appendectomy rates are no longer acceptable. AB - BACKGROUND: A 10% to 20% negative appendectomy rate has been accepted in order to minimize the incidence of perforated appendicitis with its increased morbidity. We reviewed our experience with appendicitis in order to determine the incidence of negative appendectomies and perforation, and the role of delay in diagnosis or treatment. METHODS: We reviewed 659 appendectomies performed over a 12-month period. Incidental and pediatric appendectomies were excluded. RESULTS: Seventy five percent of patients were male and 25% female. Nine percent had negative appendectomies and 28% had perforated appendicitis. Perforated appendicitis resulted in increased morbidity and length of stay. Delay in presentation greater than 12 hours after the onset of symptoms significantly increased the perforation rate. In-hospital delay did not affect perforation rate. CONCLUSIONS: We have achieved a negative appendectomy rate lower than that in other reported series, while maintaining an acceptable perforation rate. In the majority of patients, perforated appendicitis is a result of late presentation. PMID- 9409606 TI - Outcome of pouch-related complications after ileal pouch-anal anastomosis. AB - BACKGROUND: Creation of a small intestinal reservoir after ileal pouch-anal anastomosis (IPAA) results in an improved quality of life because of significantly diminished stool frequency. However, a number of complications associated with the pouch may jeopardize these sphincter-sparing procedures and occasionally result in permanent ileostomy. This study was conducted to assess the incidence, risk factors, clinical characteristics, management strategies, and outcome of pouch-related complications after IPAA. METHODS: Data on all patients undergoing IPAA with a J pouch between 1983 and Spring 1997 were prospectively gathered. Patients with pouch-specific complications were identified, and both inpatient and outpatient records analyzed in detail. When necessary, telephone contact was made to update functional data. Other parameters evaluated included age, gender, diagnosis, medication history, diagnostic modalities, laboratory values, time course, management strategies, reoperative procedures, and final results. RESULTS: Some 510 IPAA procedures were performed between 1983 and Spring 1997; 87% of patients had inflammatory bowel disease. Operative mortality was 0%. In the entire series, 27 (5.3%) had complications related to the J pouch. Of those, 22 (81%) had ulcerative colitis and were on a mean dose of 32 mg/day of prednisone. Computed tomography scan made the diagnosis in 18 (67%) and the mean white blood cell count on admission was 14,400. In 11 (41%), the complications occurred after IPAA whereas in the other 16 (59%) it occurred after ileostomy closure. In 5 (19%), the complication resolved with intravenous antibiotics and percutaneous drainage, and 22 (81%) required reoperation. Proximal (11, or 41%) and distal (8, or 30%) pouch leaks or cuff abscesses were the most common complication and accounted for 19 (70%) of the complications observed. In this series, 3 patients (11%) had complications severe enough to warrant J pouch excision, and 1 patient had a permanent ileostomy without excision. Overall pouch excision/ failure in this series was 0.78%. CONCLUSION: Complications involving the J pouch are a seemingly unavoidable part of sphincter-sparing surgery for colonic mucosal diseases. However, if therapy is timely, aggressive, and judicious for these complex patients, pouch loss should be uncommon and long-term results acceptable. PMID- 9409607 TI - Medical versus surgical management of diverticulitis in patients under age 40. AB - BACKGROUND: Diverticulitis in patients under age 40 is a distinct entity. We compared the medical versus surgical management of diverticulitis for complications and outcomes in these patients. METHODS: A retrospective review was performed for treatment, hospitalizations, complications, and outpatient visits. Complications included readmission, recurrent symptoms after antibiotic therapy, and postoperative problems. RESULTS: Twenty-nine patients had a radiographic or surgical diagnosis of diverticulitis (18 surgical, 11 medical). Medically managed patients had significantly more emergency department visits (4.7 +/- 6.6 versus 0.3 +/- 0.6, P < or =0.01), and readmissions (7 versus 4, P < or =0.02). Three surgical patients (17%) had a total of 6 complications as compared with 6 medical patients (55%) with 25 complications (chi square, P < or =0.05). All medically treated patients had recurrent symptoms, and 6 required surgery. CONCLUSION: Medically managed patients had significantly more emergency department visits and complications than those managed surgically. Surgery is the indicated treatment for the first episode of diverticulitis in patients under age 40. PMID- 9409608 TI - Mobile atheroma of the aortic arch is an underestimated source of embolization. AB - BACKGROUND: Mobile atheroma are associated with increased perioperative strokes in patients undergoing coronary artery bypass surgery. Peripheral embolization is an additional risk. Transesophageal echocardiography (TEE) accurately identifies mobile atheroma. Recent reports have discussed the possible influence of anticoagulant therapy in promoting peripheral cholesterol embolization. METHODS: Fourteen patients with mobile atheroma were treated with anticoagulation. A review of literature reporting results and complications of anticoagulation in the treatment of this condition was compared with our recent experience. RESULTS: Between 1994 and 1996, 14 patients with peripheral embolization and mobile atheroma confirmed by TEE were anticoagulated. Clinical follow-up between 6 to 30 months has demonstrated no further evidence of systemic embolization since anticoagulation. Furthermore, repeat TEE in 3 of 14 patients no longer visualized mobile atheroma. CONCLUSIONS: Mobile atheroma are recognized sources for embolization. Patients with generalized atherosclerosis should be screened for this condition in cases of systemic embolization. Anticoagulation may have therapeutic considerations in the management of this condition. PMID- 9409609 TI - Meta-analysis of the risk of metachronous hernia in infants and children. AB - BACKGROUND: Inguinal herniorrhaphy is the most common general surgical procedure performed in children. The presence of a contralateral patent processus vaginalis forms the basis of the recommendation for contralateral exploration in patients undergoing unilateral herniorrhaphy. However, a patent processus vaginalis does not necessarily go on to become a clinically apparent inguinal hernia. METHODS: All published pediatric series, in which patients underwent unilateral inguinal hernia repair and were evaluated for the development of a metachronous hernia, were included. The incidence of and risk factors associated with development a metachronous hernia were evaluated with meta-analysis. RESULTS: There were 15,310 patients ranging in age from birth to 16 years, including premature infants. Of these, 1,062 patients (7%) developed a metachronous hernia. Gender and age were not risk factors. There was an 11% risk of metachronous hernia if the original hernia was on the left side, a risk that was 50% greater than if the original hernia was on the right. Of patients who developed a metachronous hernia, 90% did so within 5 years. The complication rate of metachronous hernia was 0.5%. CONCLUSION: There is no role for routine contralateral groin exploration. High risk infants and children, especially those who undergo left inguinal herniorrhaphy, may benefit from contralateral groin exploration. If a patent processus vaginalis is found, it should be ligated. Patients who do not undergo contralateral groin exploration should be followed up for 5 years. PMID- 9409610 TI - Selective bowel decontamination in hospitalized patients awaiting liver transplantation. AB - BACKGROUND: Infection remains a major contributor to morbidity and mortality following orthotopic liver transplantation (OLT). Selective bowel decontamination (SBD) in hospitalized patients is one strategy for prophylaxis. METHODS: A retrospective case-control study was performed using 18 consecutive hospitalized patients receiving SBD prior to OLT during the period September 1995 to September 1996. Eighteen consecutive hospitalized patients without SBD transplanted during the period March 1995 to September 1995 served as a historical control group. RESULTS: Selective bowel decontamination was associated with a significantly decreased prevalence of positive cultures for gram-negative bacteria and fungi and reduced overall hospital charges. CONCLUSION: In hospitalized patients awaiting OLT, SBD is an effective prophylactic measure against infectious morbidity associated with gram-negative bacteria and fungi. PMID- 9409611 TI - Intrathyroidal parathyroid glands can be a cause of failed cervical exploration for hyperparathyroidism. AB - BACKGROUND: The incidence of intrathyroidal parathyroid glands remains controversial. The purpose of this study was to determine the incidence in a series of patients with hyperparathyroidism. METHODS: Three hundred nine patients underwent parathyroidectomy. Patients were divided into two groups: uniglandular disease versus hyperplasia. RESULTS: Eighteen of 309 patients (6%) had abnormal intrathyroidal parathyroid glands. The incidence was 3% (7 of 222) in patients with uniglandular disease versus 15% (11 of 73) in those with hyperplasia. With a mean follow-up of 54 months, 12 patients are eucalcemic, 5 have persistent hypocalcemia, and 1 has recurrent hypercalcemia. There were no recurrent laryngeal nerve injuries. CONCLUSIONS: These data suggest that an intrathyroidal adenoma is an uncommon cause of failure, whereas abnormal intrathyroidal parathyroid tissue may be a more common cause of failure in patients with hyperplasia. PMID- 9409612 TI - Risk factors in patients undergoing major nonvascular abdominal operations that predict perioperative myocardial infarction. AB - BACKGROUND: Perioperative myocardial infarction (PMI) is an uncommon but serious complication of major abdominal surgery. Identifying the patients at risk may potentially reduce morbidity and mortality. In this study we determined risk factors associated with PMI in patients undergoing abdominal, nonvascular surgery (ANVS). METHODS: The utility of risk factors for PMI using Goldman's criteria and nine other variables were compared in patients diagnosed with PMI after ANVS (group I) and a control group (group II) matched for age, gender, and type of operation. RESULTS: Thirty-four patients, 21 men and 13 women, with a mean age of 70 years were diagnosed with PMI, which was associated with a 41% mortality rate (14 of 34). Risk factors for PMI included poor general condition, congestive heart failure, abnormal cardiac rhythm, smoking, previous myocardial infarction (MI), and emergent operation. CONCLUSION: Although PMI following ANVS is uncommon, the mortality rate remains high. Patients classified as Goldman's class III and IV, or with a history of cigarette smoking, previous MI, or angina merit further evaluation in order to reduce the incidence of this complication. PMID- 9409613 TI - Retransplantation in the diabetic patient with a pancreas allograft. AB - BACKGROUND: Retransplantation has been considered a risk factor for both postoperative complications and diminished graft survival, especially in diabetic patients. METHODS: A retrospective survey was performed of a consecutive case series of 196 pancreas transplants in 186 diabetic patients. All patients underwent whole organ pancreas transplantation with bladder drainage. RESULTS: A total of 33 pancreas transplants (17%) in 30 patients were performed after previous transplant. The mean interval between transplants was 3.9 years. At the time of retransplantation, 16 patients had concomitant procedures. Venous extension grafts were used in 10 patients. The mean length of initial hospital stay was 19.5 days, and mean hospital charges were approximately $125,000. The incidences of rejection, infection, and operative complications were 61%, 67%, and 45%, respectively. Patient survival was 90%, kidney graft survival was 82%, and pancreas graft survival was 61% after a mean follow-up of 29 months. Complete rehabilitation was achieved in 73% of cases. CONCLUSIONS: Pancreas transplantation after previous transplant is a challenging but safe treatment that often requires concomitant procedures, the use of vascular extension grafts, and atypical placement of the allograft. However, the good results justify an aggressive policy of retransplantation in the diabetic patient either with a failed allograft or functioning kidney transplant. PMID- 9409614 TI - Crosslinking of the U5 snRNP-specific 116-kDa protein to RNA hairpins that block step 2 of splicing. AB - Step 2 of pre-mRNA splicing has characteristics that are suggestive of a 5' to 3' scanning process from the branch point to locate the 3' splice site. Specifically, the 3' splice site is almost always at the first AG downstream of the branch point even when the two elements are separated by hundreds of nucleotides. Insertion of new AGs between the branch and 3' splice site, or mutation of the wild-type 3' splice site, usually results in use of the new first AG as the 3' splice site. Finally, insertion of stable secondary structure between the branch point and 3' splice site, but distant from both elements, results in a block to step 2. We have sought to complement this circumstantial evidence by detecting physical contacts between the spliceosome and the RNA substrate in regions that are not themselves important for splicing, other than that they lie between the branch point/polypyrimidine tract and the 3' splice site. We have blocked step 2 of splicing by insertion of hairpin structures between the branch point and 3' splice site and applied methylene blue-mediated crosslinking, which is specific for protein-dsRNA interactions. Using this approach, we have detected a 116-kDa crosslinked protein that appears after step 1 of splicing with all transcripts containing a hairpin downstream of the branch point. The protein was identified as the 116-kDa U5 snRNP protein, which is a GTP binding protein involved in step 2 of splicing. The crosslinking characteristics of U5 p116 are consistent with it having a role in locating the 3' splice site AG prior to step 2 of splicing. PMID- 9409615 TI - Intragenic suppression in tRNA: evidence for crosstalk between the D and the T stems. AB - We showed previously that introduction of two of the three unique features of Escherichia coli initiator tRNA onto an elongator methionine tRNA conferred significant activity in initiation. Surprisingly, introduction also of the third unique feature, the A11:U24 base pair in the D stem, resulted in total lack of accumulation of the mutant Mi:3 tRNA. We show here that the Mi:3 tRNA gene is transcribed efficiently in vitro. Processing of the Mi:3 precursor transcript shows, however, that both the precursor and the mature Mi:3 tRNA are unstable in E. coli extracts. To understand the basis of instability caused by the A11:U24 base pair in the elongator methionine tRNA background, we have isolated and characterized intragenic suppressor mutations in the tRNA that restore its function in translation initiation. Sequence changes in the T stem that convert the existing A51 x C63 mismatch to a base pair in the Mi:3 tRNA result in accumulation of the tRNAs in vivo. The initiation activity and in vivo levels of accumulation of these suppressors are in the order Mi:3/G51:C63 > Mi:3/A51:U63 >> Mi:3/G51.U63. These results show that the in vivo accumulation of a tRNA with A11:U24 base pair in the D stem depends upon a base pair between positions 51 and 63 in the T stem. Structural analysis in vitro of the Mi:3 and Mi:3/G51:C63 transcripts suggests that the Mi:3 tRNA is unable to adopt a stable tRNA-like conformation. Various considerations suggest that this is most likely due to a high entropic barrier to tertiary interactions, between the D and the T loops necessary for the formation of a stable tRNA structure. PMID- 9409616 TI - Purification and cDNA cloning of the AdoMet-binding subunit of the human mRNA (N6 adenosine)-methyltransferase. AB - The methylation of internal adenosine residues in eukaryotic mRNA, forming N6 methyladenosine (m6A), is catalyzed by a complex multicomponent enzyme. Previous studies suggested that m6A affects the efficiency of mRNA processing or transport, although the mechanism by which this occurs is not known. As a step toward better understanding the mechanism and function of this ubiquitous posttranscriptional modification, we have shown that HeLa mRNA (N6-adenosine) methyltransferase requires at least two separate protein factors, MT-A and MT-B, and MT-A contains the AdoMet binding site on a 70-kDa subunit (MT-A70). MT-A70 was purified by conventional chromatography and electrophoresis, and was microsequenced. The peptide sequence was used to design a degenerate oligodeoxynucleotide that in turn was used to isolate the cDNA clone coding for MT-A70 from a HeLa cDNA library. Recombinant MT-A70 was expressed as a fusion protein in bacteria and was used to generate anti-MT-A70 antisera in rabbits. These antisera recognize MT-A70 in HeLa nuclear extracts by western blot and are capable of depleting (N6-adenosine)-methyltransferase activity from HeLa nuclear extract, confirming that MT-A70 is a critical subunit of (N6-adenosine) methyltransferase. Northern blot analysis reveals that MT-A70 mRNA is present in a wide variety of human tissues and may undergo alternative splicing. MT-A70 cDNA probe hybridizes to a 2.0-kilobase (kb) polyadenylated RNA isolated from HeLa cells, whereas it hybridizes to two predominant RNA species (approximately 2.0 kb and 3.0 kb) using mRNA isolated from six different human tissues. Analysis of the cDNA sequence indicates that it codes for a 580-amino acid protein with a predicted MW = 65 kDa. The predicted protein contains sequences similar to consensus methylation motifs I and II identified in prokaryotic DNA (N6 adenosine)-methyltransferases, suggesting the functional conservation of peptide motifs. MT-A70 also contains a long region of homology to the yeast protein SPO8, which is involved in induction of sporulation by an unknown mechanism. PMID- 9409617 TI - Natural base-pairing interactions between 5' splice site and branch site sequences affect mammalian 5' splice site selection. AB - In the murine gene encoding the neuronal cell adhesion molecule (NCAM), the integrity of the 5' splice site of exon 18 (E18) is essential for regulation of alternative splicing. To further study the contribution of 5' splice site sequences, we used a simple NCAM pre-mRNA containing a portion of E18 fused to E19 and separated by a shortened intron. This RNA is spliced in vitro to produce five sets of lariat intermediates and products, the most abundant set displaying aberrant migration in acrylamide/urea gels. Base pairing interactions between positions +5 and +8 of the intron and positions -3 and -6 from the branch point were responsible for the faster migration of this set of lariat molecules. To test whether the duplex structure forms earlier and contributes to 5' splice site selection, we used NCAM substrates carrying the 5' splice sites of E17 and E18 in competition for the 3' splice site of E19. Mutations upstream of the major branch site improve E18/E19 splicing in NIH3T3 extracts, whereas compensatory mutations at positions +7 and +8 neutralize the effect of branch site mutations and curtail E18/E19 splicing. Our data suggest that duplex formation occurs early and interferes with the assembly of complexes initiated on the 5' splice site of NCAM E18. This novel type of intron interaction may exist in the introns of other mammalian pre-mRNAs. PMID- 9409618 TI - Identification of a minimal Alu RNA folding domain that specifically binds SRP9/14. AB - We have identified functionally and analyzed a minimal Alu RNA folding domain that is recognized by SRPphi14-9. Recombinant SRPphi14-9 is a fusion protein containing on a single polypeptide chain the sequences of both the SRP14 and SRP9 proteins that are part of the Alu domain of the signal recognition particle (SRP). SRPphi14-9 has been shown to bind to the 7SL RNA of SRP and it confers elongation arrest activity to reconstituted SRP in vitro. Alu RNA variants with homogeneous 3' ends were produced in vitro using ribozyme technology and tested for specific SRPphi14-9 binding in a quantitative equilibrium competition assay. This enabled identification of an Alu RNA of 86 nt (SA86) that competes efficiently with 7SL RNA for SRPphi14-9 binding, whereas smaller RNAs did not. The secondary structure of SA86 includes two stem-loops that are connected by a highly conserved bulge and, in addition, a part of the central adaptor stem that contains the sequence at the very 3' end of 7SL RNA. Circularly permuted variants of SA86 competed only if the 5' and 3' ends were joined with an extended linker of four nucleotides. SA86 can thus be defined as an autonomous RNA folding unit that does not require its 5' and 3' ends for folding or for specific recognition by SRPphi14-9. These results suggest that Alu RNA identity is determined by a characteristic tertiary structure, which might consist of two flexibly linked domains. PMID- 9409619 TI - A unique intronic splicing enhancer controls the inclusion of the agrin Y exon. AB - Alternative splicing of the agrin mRNA controls the ability of agrin protein to induce the clustering of acetylcholine receptors at the neuromuscular junction. Using a transfectable reporter gene, we show that one agrin alternative exon, the Y exon, is controlled by a regulatory sequence in the downstream intron. Portions of this intronic sequence have the properties of a splicing enhancer that can activate splicing of a heterologous exon when placed in the intron downstream. The regulatory region is complex in structure, containing several different elements capable of activating splicing. Individual enhancing elements differ in their cell-type specificity, and are not apparently synergistic, as two elements together induce lower splicing than either does separately. Essential nucleotides within these regulatory elements were identified by scanning mutagenesis across the active region. Interestingly, the elements do not appear similar to known intronic splicing enhancer elements. This Y exon enhancer and its components take part in an apparent combinatorial system of control where multiple regulatory elements of varying activity combine to produce a precisely cell-specific exon inclusion. As a major contributor to the regulation of the Y exon, the enhancer ultimately controls the properties of the agrin protein. PMID- 9409620 TI - Ni2+-binding RNA motifs with an asymmetric purine-rich internal loop and a G-A base pair. AB - RNA molecules with high affinity for immobilized Ni2+ were isolated from an RNA pool with 50 randomized positions by in vitro selection-amplification. The selected RNAs preferentially bind Ni2+ and Co2+ over other cations from first series transition metals. Conserved structure motifs, comprising about 15 nt, were identified that are likely to represent the Ni2+ binding sites. Two conserved motifs contain an asymmetric purine-rich internal loop and probably a mismatch G-A base pair. The structure of one of these motifs was studied with proton NMR spectroscopy and formation of the G-A pair at the junction of helix and internal loop was demonstrated. Using Ni2+ as a paramagnetic probe, a divalent metal ion binding site near this G-A base pair was identified. Ni2+ ions bound to this motif exert a specific stabilization effect. We propose that small asymmetric purine-rich loops that contain a G-A interaction may represent a divalent metal ion binding site in RNA. PMID- 9409621 TI - Inhibition of mammalian spliceosome assembly and pre-mRNA splicing by peptide inhibitors of protein kinases. AB - Four peptides are shown to block mammalian spliceosome assembly and pre-mRNA splicing in vitro. Previously, these peptides have been shown to inhibit Ca2+ dependent calmodulin kinase II (CaMK II) via distinct mechanisms. One is a competitive inhibitor of the kinase, two interfere with autophosphorylation events, and one competes for binding to calmodulin, a CaMK II-activating protein. However, because EGTA does not inhibit splicing, the involvement of CaMK II itself in splicing is unlikely; rather, a protein similar to CaMK II may be involved in spliceosome assembly and splicing. Two of the inhibitory peptides, the calmodulin binding domain (CBD) and glycogen synthase (GS) fragment, block assembly of spliceosomal complex C. These peptides inhibited splicing if they were added to reactions any time within the first 10 min of splicing assays. No inhibition of spliceosome assembly or splicing occurred in the presence of randomized versions of the CBD or GS peptide. Additionally, the GS peptide inhibited splicing when added to assays at later time points, despite the fact that spliceosomal complex C had formed. Cumulatively, these analyses suggest that the peptides inhibit at least two distinct events in the spliceosomal cycle. The first event occurs early during in vitro splicing. For this event, prolonged incubations of splicing reactions do not result in a recovery of splicing activity. The second event occurs later and represents a slowing of an essential step, because splicing activity can be recovered in prolonged incubations. Peptides known to inhibit protein kinase A and protein kinase C had no effect on pre-mRNA splicing, underscoring the specificity of the observed inhibitory effects. PMID- 9409622 TI - The human U5 snRNP-specific 100-kD protein is an RS domain-containing, putative RNA helicase with significant homology to the yeast splicing factor Prp28p. AB - Through UV-crosslinking experiments, we previously provided evidence suggesting that a U5 snRNP protein with a molecular weight in the 100-kDa range is an ATP binding protein (Laggerbauer B, Lauber J, Luhrmann R, 1996, Nucleic Acid Res 24:868-875). Separation of HeLa U5 snRNP proteins on 2D gels revealed multiple variants with apparent molecular masses of 100 kDa. Subsequent microsequencing of these variants led to the isolation of a cDNA encoding a protein with an N terminal RS domain and a C-terminal domain that contains all of the conserved motifs characteristic of members of the DEAD-box family of RNA-stimulated ATPases and RNA helicases. Antibodies raised against cDNA-encoded 100-kDa protein specifically recognized native U5-100kD both on immunoblots and in purified HeLa U5 snRNPs or [U4/U6.U5] tri-snRNP complexes, confirming that the bona fide 100 kDa cDNA had been isolated. In vitro phosphorylation studies demonstrated that U5 100kD can serve as a substrate for both Clk/Sty and the U1 snRNP-associated kinase, and further suggested that the multiple U5-100kD variants observed on 2D gels represent differentially phosphorylated forms of the protein. A database homology search revealed a significant degree of homology (60% similarity, 37% identity) between the Saccharomyces cerevisiae splicing factor, Prp28p, which lacks an N-terminal RS domain, and the C-terminal domain of U5-100kD. Consistent with their designation as structural homologues, anti-Prp28 antibodies recognized specifically the human U5-100kD protein on immunoblots. Together with the DEXH box U5-200kD protein (Lauber J et al., 1996, EMBO J 15:4001-4015), U5-100kD is the second example of a putative RNA helicase that is tightly associated with the U5 snRNP. Given the recent identification of the U5-116kD protein as a homologue of the ribosomal translocase EF-2 (Fabrizio P, Laggerbauer B, Lauber J, Lane WS, Luhrmann R, 1997, EMBO J 16:4092-4106), at least three integral U5 snRNP proteins thus potentially facilitate conformational changes in the spliceosome during nuclear pre-mRNA splicing. PMID- 9409623 TI - tRNA recognition for modification: solution probing of tRNA complexed with Escherichia coli tRNA (guanosine-1) methyltransferase. AB - The interaction of Escherichia coli tRNA (guanosine-1) methyltransferase and tRNA(1Leu) transcripts has been probed using cleavage with iodine of phosphorothioate-substituted transcripts, lead acetate, and enzymes specific for single- and double-stranded RNA. All lytic agents protect the anticodon stem-loop and variable loop regions against cleavage, and some protection is also seen in core structures of the tRNA. Residues from both strands of the anticodon stem are protected against cleavage with iodine and lead by enzyme, yet positions G37 and G36, which are crucial for catalysis and binding, are not. This suggests that these residues may undergo structural perturbation in the presence of S-adenosyl methionine. Occupancy of the AdoMet site by the product S-adenosyl-homocysteine, a potent inhibitor of the enzyme, has little or no effect on tRNA binding or protection. Enhanced reactivity with lead is seen at residues located in the anticodon stem-loop, extra-loop, and core (C34, U47c, and G49), which suggests some perturbations in RNA structure might accompany binding. PMID- 9409624 TI - Polyadenylation of telomerase RNA in budding yeast. AB - Telomerase RNA is a subunit of a stable ribonucleoprotein particle required for telomere replication. We find that, at steady state, 5-10% of the telomerase RNA in Saccharomyces cerevisiae and Kluyveromyces lactis contains a poly(A) tail of about 80 nt. In S. cerevisiae, the poly(A)+ fraction quickly disappeared when a conditional pap1 or rna15 mutant was shifted to the nonpermissive temperature, indicating that polyadenylation is accomplished by the same machinery that polyadenylates mRNAs. Potential cis-acting polyadenylation elements were identified in the telomerase RNA sequence; when they were mutagenized, the polyadenylation pattern shifted, but was not eliminated. The corresponding mutants displayed wild-type growth. By putting the RNA under the control of an inducible promoter, we were able to show that synthesis of the poly(A)+ RNA precedes that of the poly(A)- fraction. This supports, but does not prove, a model in which all telomerase RNA is first polyadenylated and then rapidly processed to give the stable poly(A)-form. Cell cycle arrest experiments showed an increase in the poly(A)+ form between G1 and S phase, consistent with an induction of telomerase RNA transcription at the time of DNA replication. PMID- 9409625 TI - Synthesis of 3'-thioribonucleosides and their incorporation into oligoribonucleotides via phosphoramidite chemistry. AB - Oligoribonucleotides containing 3'-S-phosphorothiolate linkages are valuable probes in nucleic acid biochemistry, but their accessibility has been limited because 3'-thioribonucleoside phosphoramidites have not been available. We synthesized 3'-thioribonucleoside derivatives (C, G, and U) via glycosylations of nucleoside bases with 3-S-thiobenzoyl-5-O-toluoyl-1,2-O-diacetylfuranose 5, which was obtained from 1 ,2-O-isopropylidene-5-O-toluoyl-3-trifluoromethane-sulfonyl alpha-D-x ylofuranose 2 by SN2 displacement with sodium thiobenzoate. Additionally, a 3'-thioinosine derivative was prepared from inosine via direct modification of the ribose, analogous to the previously reported synthesis of 3' thioadenosine, except that the intermediate 2',3'-epoxide 9 was first protected as the 5'-O-tert-butyldiphenylsilyl ether prior to subsequent synthetic steps. This hydrophobic silyl group facilitated extraction and isolation of synthetic intermediates. After removal of the protecting groups, the 3'-thionucleosides (C, G, U, and I) were treated with 2,2'-dipyridyl disulfide to protect the free thiol group as a disulfide. The 3'-thionucleosides were converted to the corresponding phosphorothioamidites using procedures analogous to those for standard phosphoramidites. The amino groups of 3'-thiocytidine and 3'-thioguanosine were protected as benzoyl and isobutyryl amides, respectively, and the 5'- and 2' hydroxyl groups of each nucleoside were protected as dimethoxytrityl and tert butyldimethylsilyl ethers, respectively. The 3'-thiol group was deprotected by reduction with DTT and phosphitylated to afford analytically pure 3'-S phosphorothioamidites 15, which were incorporated into oligoribonucleotides by solid-phase synthesis. Chemical assays and mass spectrometry of the synthetic RNA showed that ribose-3'-S-phosphorothiolate linkages were installed correctly and efficiently into RNA oligonucleotides using phosphoramidite chemistry. PMID- 9409626 TI - Acrylic acid: two-generation reproduction toxicity study in Wistar rats with continuous administration in the drinking water. AB - In a two-generation reproduction toxicity study, groups of 25 male and 25 female Wistar rats (for both F0 and F1 generations) received acrylic acid (AA) in the drinking water at concentrations of 0 (control), 500, 2500 and 5000 ppm for at least 70 days prior to mating, through mating, gestation, lactation and to weaning. The study continued through to weaning of the F2 offspring at 21 days of age. Achieved intakes of AA for the F0 and F1 parents during premating ranged from 46 (500 ppm) to 502 (5000 ppm) mg/kg/day. AA had no adverse effects on fertility and reproductive performance of the parent rats at doses up to 5000 ppm. General systemic toxicity was apparent with reduced body weights, food and water consumption in F0 parents at 5000 ppm and in F1 parents at 2500 and 5000 ppm; the only treatment-related pathological finding was a minimal hyperkeratosis of the limiting ridge of the forestomach with a minimal oedema of the submucosa of the glandular stomach in both parental generations at 5000 ppm. Dose-related signs of developmental toxicity were detected in F1 and F2 pups at 2500 and 5000 ppm in the form of retarded growth and some delay in the eye/auditory canal opening in F2 pups, but there was no evidence that AA had an adverse influence on pup morphology. Thus, the no-observed-adverse-effect level (NOAEL) is 5000 ppm for fertility and reproductive performance of the parents, 2500 ppm (F0 parents) or 500 ppm (F1 parents) for general systemic toxicity and 500 ppm for developmental toxicity. PMID- 9409627 TI - Developmental toxicity study of inhaled acrylic acid in New Zealand White rabbits. AB - In range-finding and definitive developmental toxicity studies, timed pregnant New Zealand White rabbits were exposed to acrylic acid (CAS No. 79-10-7) vapour for 13 consecutive days during pregnancy. In the range-finding study, eight pregnant does/group were exposed to 30, 60, 125 or 250 ppm acrylic acid vapour on gestation days (gd) 10-22 of pregnancy. Monitors of toxicity included body weight measurements, daily food consumption measurements and clinical observations. Three of the eight does/group were killed on the day following the last exposure (gd 23), and the remaining does were killed and autopsied on gd 29. At autopsy, special attention was given to gross observation of maternal nasal turbinates, and nasal turbinates from all does were evaluated histologically. No evaluation of foetuses was performed in the range-finding study. In the definitive study, 16 does/group were exposed to concentrations of 25, 75 or 225 ppm acrylic acid vapour from gd 6 to 18, the major period of organogenesis. Monitors of maternal toxicity included clinical observations and measurements of body weight and daily food consumption measurements. Does were killed and autopsied on gd 29. Maternal liver and kidney weights were measured and external, visceral and skeletal evaluations of foetuses were conducted. Maternal nasal turbinates were not evaluated histologically in the definitive study. Effects in does from both studies included consistent concentration-related reductions in food consumption and body weight gains throughout the exposure period at concentrations of acrylic acid vapour above 60 ppm. Characteristic clinical signs of sensory irritation, including perinasal and perioral wetness and severe nasal congestion, were noted in does from both studies at or above vapour concentrations of 75 ppm. Gross observation of nasal turbinates immediately following exposures in the range finding study indicated colour changes in the nasal turbinates of does in the 60 and 250 ppm groups. Colour changes in the nasal turbinates were noted in one doe from the 250 ppm exposure group killed on gd 29. Pertinent autopsy findings in the does from the definitive study included ulceration of the nasal turbinates of a single doe in the 225 ppm group. Histological evaluation of turbinates from does killed the day following exposures in the range-finding study revealed lesions in the nasal epithelium in all acrylic acid-exposed groups. The severity of the lesions was concentration related. Microscopic evaluation of turbinates from does killed on gd 29 showed the presence of nasal lesions in the 60, 125 and 250 ppm groups. However, the nasal tissues had recovered considerably during the post-exposure interval. Despite the severe effects on the nasal mucosa of does in both studies, there was no evidence of developmental toxicity including teratogenicity at any exposure concentration used in the definitive study. PMID- 9409629 TI - Cadmium accumulation in liver and kidney of mice exposed to the same weekly cadmium dose continuously or once a week. AB - Cd levels in blood, liver and kidney of female mice were measured after exposure to Cd as CdCl2 in the food, either continuously (CE group) throughout the week (300 microg Cd/kg feed) or for 24 hr/wk (2100 microg Cd/kg) for 5 wk (occasionally exposed, OE group). In a control group that received feed with Cd levels below the detection limit (< 7 microg/kg), Cd levels in blood, liver and kidneys were below the detection limit after the 5 wk of exposure. The weekly dose of Cd administered to the exposed CE and OE groups was similar (approx. 400 microg Cd/kg mice/wk). The OE group had a higher Cd level in blood and a higher fractional accumulation (% of dose) of Cd in the liver and kidneys compared with the CE group. This indicates that the fractional Cd absorption in the gastrointestinal tract is higher when high Cd doses are ingested occasionally than when low doses are ingested continuously, even if weekly doses are the same. It is hypothesized that this difference in absorption could be due to Cd-induced unspecific damage to the intestinal mucosa, changes in tight-junction permeability caused by Cd, or to a saturation of the Cd-binding capacity of the intestinal mucosa in mice exposed to high Cd levels occasionally. PMID- 9409628 TI - Testing the potential of sodium fluoride to affect spermatogenesis in the rat. AB - The potential of sodium fluoride (NaF) to affect spermatogenesis and endocrine function was assessed in P and F1 generation male rats. Male and female experimental rats received sodium fluoride in their drinking water at one of four concentrations (25, 100, 175, 250 ppm). P generation male and female rats were exposed to sodium fluoride in their drinking water for 10 wk and then males were mated to females within the same treatment groups. Reproductive tissues were collected from P generation male rats after approximately 14 wk of treatment. Pregnant females (P) were exposed to sodium fluoride via their drinking water through gestation and lactation. F1 generation weanling male rats remained within the same treatment groups as their parents. F1 generation male rats were exposed to sodium fluoride in their drinking water for 14 wk, at which time reproductive tissues were collected. Dose-related effects were not observed within the P and F1 treatment groups in testis weights, prostate/seminal vesicle weights, non reproductive organ weights, testicular spermatid counts, sperm production per gram of testis per day, sperm production per gram of testis, LH, FSH or serum testosterone concentrations. Histological changes were not observed in testicular tissues from either the P or F1 generation. We conclude that prolonged exposure to sodium fluoride in drinking water at the doses administered in this study does not adversely affect spermatogenesis or endocrine function in the P and F1 generation male rats. PMID- 9409630 TI - Mutagenicity of azo dyes used in foods, drugs and cosmetics before and after reduction by Clostridium species from the human intestinal tract. AB - Various azo dyes currently approved by the US FDA for use in foods, drugs and cosmetics are reduced by anaerobic bacteria from the human intestinal tract. These bacteria with azoreductase activities include several Clostridium species. Seven of these azo dyes and their reduction products following incubation with a Clostridium sp. were evaluated for mutagenicity in Salmonella typhimurium strains TA98 and TA100. No mutagenicity was induced in either TA98 or TA100 by any of the seven azo dyes or the reduced metabolites when tested at concentrations as high as 200 microg/plate, with or without exogenous metabolic activation by rat liver fraction S-9. PMID- 9409631 TI - A single amino acid substitution (Leu160His) in cytochrome P450 CYP2A6 causes switching from 7-hydroxylation to 3-hydroxylation of coumarin. AB - Human populations are thought to metabolize coumarin almost exclusively by 7 hydroxylation. We have identified an individual who is homozygous for a single amino acid substitution (Leu160His) in the cytochrome P450 CYP2A6 arising from the variant CYP2A6*2 allele. On administration of coumarin (2 mg orally) no detectable 7-hydroxycoumarin was excreted in the 0-8-hr urine, rather, approximately 50% of the dose was eliminated as 2-hydroxyphenylacetic acid, the end-product of coumarin 3-hydroxylation. His immediate family members, who were heterozygous for the CYP2A6*2 allele, excreted little 2-hydroxyphenylacetic acid and mainly 7-hydroxycoumarin, when similarly tested. These findings raise a question regarding human risk evaluations for environmental coumarin exposures, since 7-hydroxylation is regarded as a detoxication pathway, but 3-hydroxylation as the process required to lead to macromolecular covalent binding of coumarin. Persons homozygous for the CYP2A6*2 allele may constitute 1-25% of various populations. PMID- 9409632 TI - Evaluation of chronic oral toxicity and carcinogenic potential of lornoxicam in rats. AB - As part of the preclinical development program for lornoxicam, a novel non steroidal anti-inflammatory drug (NSAID), its chronic oral toxicity and carcinogenic potential was assessed in Sprague-Dawley rats. Male and female rats were administered lornoxicam by oral gavage at 0, 0.06, 0.16 or 0.40 mg/kg/day for 12 months or at 0, 0.01 or 0.06 mg/kg/day in a supplementary low-dose study of the same duration (main group: 20/sex/group; 4-wk recovery: five/sex/group; satellites for electrocardiography and toxicokinetics: five/sex/group). Drug related toxicity mainly comprised mortality, reduced body weight gain, clinico pathological changes indicative of anaemia resulting from blood loss, and renal damage, renal papillary necrosis and gastrointestinal mucosal lesions. The kidney associated changes were not completely reversible during the recovery period. Toxicokinetic investigations demonstrated a dose-linear absorption of the drug. In female rats the terminal half-life was about twice that in males which led to a higher exposure of this gender to lornoxicam. A dose of 0.01 mg/kg/day was established as no-observed-effect level. In a 104-wk carcinogenicity study, lornoxicam was administered by oral gavage to male and female rats (50/sex/group) at 0 (control 1), 0 (control 2), 0.0625, 0.125 or 0.250 mg/kg/day. In females only, the high dose was reduced twice during the study due to toxicity observed (0.250 to 0.200 to 0.160 mg/kg/day). Drug-related changes were similar to those in the chronic studies and consistent with the anticipated side-effects of NSAIDs. No carcinogenic potential was revealed. PMID- 9409633 TI - Skin sensitization testing: the relevance of rechallenge and pretreatment with sodium lauryl sulfate in the guinea pig maximization test. AB - The guinea pig maximization test is one of the preferred test methods for the identification of skin sensitizers. The OECD/EC test guidelines allow for the conduct of a rechallenge in case doubtful reactions are obtained after challenge. The relevance of rechallenging was investigated by performing multiple challenges (up to four) in the maximization test with four well-known sensitizers of varying strength: nickel sulfate, sulfathiazole, benzocaine, and 1-chloro-2,4 dinitrobenzene. In addition, the effect of sodium lauryl sulfate (SLS) pretreatment during topical induction with weak sensitizers on rechallenging was investigated. In contrast to what has frequently been hypothesized, rechallenge did not result in an increase of skin reaction as compared with the reactions observed after the first treatment. SLS pretreatment was very effective in increasing the initial challenge response to weak sensitizers. Subsequent rechallenging in these cases however again showed a decrease in sensitivity of the animals. PMID- 9409634 TI - Humoral immune response within the lung in HIV-1 infection. AB - In order to study the humoral immune defences in the respiratory tract during HIV 1 infection, we measured the levels, local productions and anti-HIV and antibacterial activities of IgG and IgA in the bronchoalveolar lavage fluid (BALF) and serum of 61 adult patients with severe HIV infection and of 56 HIV- controls. Albumin was used as the serum transudation factor. The increase of immunoglobulin levels in the serum of HIV-infected patients was confirmed. The IgG level was also increased in epithelial lining fluid (ELF), whereas the total IgA level was unchanged and secretory IgA (SIgA) level was decreased. The ELF/serum immunoglobulin ratios suggested that the IgG present in ELF resulted mainly from transudation, in contrast to SIgA, which was synthesized locally in controls but greatly diminished in HIV-infected patients. IgG to HIV-1 could be detected in BALF of all the patients, but IgA to HIV-1 only in 30% of patients. BAL IgG reacted more consistently and with a broader array of HIV-1 antigens than did IgA. BAL IgA, when present in samples, reacted primarily with viral envelope antigens. Because IgA specificities to some HIV-1 antigens were detected more intensively by BAL than by serum immunoglobulins, we conclude that the mucosal immune response is distinct from that in serum. IgG antibody activity to Streptococcus pneumoniae was decreased in HIV-infected patients' sera, and IgA antibody activities to S. pneumoniae and to Pseudomonas aeruginosa were decreased in ELF in HIV-infected patients. PMID- 9409635 TI - Infection with rat cytomegalovirus (CMV) in the immunocompromised host is associated with the appearance of a T cell population with reduced CD8 and T cell receptor (TCR) expression. AB - Infection with human cytomegalovirus (HCMV) mostly results in a chronic subclinical infection; the immune system is unable to eliminate the virus and is apparently in equilibrium with the persistent virus. In the immunosuppressed host this equilibrium is disturbed, resulting in clinical infection. Rat cytomegalovirus (RCMV) infection in its host can be used as a model for HCMV infection. Using flow cytometry we examined the effect of acute RCMV infection on the composition of leucocyte subsets in the peripheral blood of both immunocompetent and immunosuppressed (5 Gy total body irradiation) Lewis rats. Special attention was paid to the natural killer (NK) cells and the CD8+ T cells known to be involved in the control of viral infections. Furthermore, we determined the presence of leucocyte subsets in the internal organs by immunohistochemistry. In immunocompetent rats, infection caused a small increase in NK cells and a large increase in CD8+ T cells. In contrast, infection of immunosuppressed rats caused a marked increase in NK cells and a small increase in CD8+ T cells, consisting of T cells with reduced expression of both CD8 and TCR. This phenomenon is characteristic of anergic CD8+ T cells, possibly explaining the ability of the virus to escape elimination by the immune system. The increase of NK cells in the peripheral blood of immunosuppressed, RCMV infected rats could also be detected in kidney, liver, lung and pancreas, but not in salivary gland. This could explain the long persistence of infectious virus in the salivary gland. PMID- 9409636 TI - Antibodies protect mice against challenge with tick-borne encephalitis virus (TBEV)-infected macrophages. AB - TBEV is a flavivirus highly pathogenic for humans. By transfer of antibodies directed to the TBEV surface glycoprotein E into mice, immune protection against subsequent inoculation with free TBEV particles could be achieved. After natural TBEV infection via the skin, however, cells of the monocyte/macrophage lineage were recently demonstrated to represent an important source of local virus replication before viraemia occurs. Whether antibodies can protect against virus challenge when contracted in the form of infected cells, however, is still unclear. In the current study, TBEV antibodies protected mice against challenge with either free virus or TBEV-infected macrophages equally well. This observation may be of more general significance. PMID- 9409638 TI - Inhibition of nitric oxide synthase (NOS) aggravates Staphylococcus aureus septicaemia and septic arthritis. AB - The aim of this study was to assess the role of NO and its metabolites in bacterial arthritis. The murine model of haematogenously acquired septic arthritis was used. Swiss mice treated with NOS inhibitors (N(G)-monomethyl-L arginine or N(omega)-nitro-L-arginine methyl ester) were injected intravenously with toxic shock syndrome toxin-1 (TSST-1) producing Staphylococcus aureus LS-1. Arthritis was evaluated clinically and histopathologically. Serum cytokine levels, bacterial isolates and intracellular capacity of macrophages to kill bacteria were also analysed. The frequency of arthritis in mice treated with NOS inhibitors was three to four-fold higher than that in non-treated controls (75% versus 20%). The severity of arthritis, expressed as mean arthritic index, was 1.4 and 0.4, respectively. Cartilage and/or bone destruction occurred in 63% of NOS inhibitor-treated mice, but only in 10% of controls. Also, the cumulative septicaemia-induced mortality was clearly higher in mice treated with NOS inhibitors compared with non-treated controls. Intracellular killing capacity of the peritoneal macrophages, treated in vitro with NOS inhibitors, was decreased. Thus, 24 h after bacterial inoculation peritoneal macrophages pretreated with NOS inhibitors killed more than 10 times less bacteria than the control ones (P<0.01). We conclude that NOS inhibitors aggravate S. aureus arthritis, possibly by inducing impairment of the intracellular killing capacity of macrophages. PMID- 9409637 TI - Interferon-gamma (IFN-gamma) down-regulates the rhinovirus-induced expression of intercellular adhesion molecule-1 (ICAM-1) on human airway epithelial cells. AB - Human rhinoviruses (HRV) are a major cause of upper respiratory tract infections in man, and can exacerbate existing pulmonary disease. The major group of HRV attach to ICAM-1, which is expressed on nasal and bronchial epithelial cells. To study the influence of biological mediators on ICAM-1 expression, and consequently HRV attachment and infection, we compared the effects of various cytokines, alone and in combination, on ICAM-1 expression by an uninfected and HRV-infected bronchial epithelial cell line H292. Cytokines known to be released soon after viral infection, such as tumour necrosis factor-alpha (TNF-alpha), IL 1beta and the chemokine IL-8 increase ICAM-1 expression on uninfected cells. Epithelial cells infected with live HRV-14 displayed marked up-regulation of ICAM 1 compared with baseline. TNF-alpha further enhanced the HRV-induced increase in ICAM-1 expression on epithelial cells, peaking at day 4 after infection, whilst IL-8 exhibited a steady increase in ICAM-1 expression over 14 days. In contrast, IFN-gamma, a known Th1 antiviral lymphokine, whilst increasing the level of ICAM 1 on uninfected cells, induced a significant persistent down-regulation of ICAM-1 expression on HRV-infected epithelial cells. With combinations of TNF-alpha and IFN-gamma, ICAM-1 expression on HRV-infected cells was reduced to basal levels. The effects of IFN-gamma were paralleled by a reduction in viral titres. Our in vitro model has provided useful insights into the early pathogenic events of HRV infection at the level of the host cell-virus interaction. Our data confirm that biological mediators play a crucial role in the pathogenesis as well as the course of HRV infection which is modulated by the types, and time kinetics of inflammatory cytokines in the immediate microenvironment. PMID- 9409639 TI - Trypanosoma cruzi induces strong IL-12 and IL-18 gene expression in vivo: correlation with interferon-gamma (IFN-gamma) production. AB - IFN-gamma, produced after infection with Trypanosoma cruzi, has been shown to be crucial in the determination of resistance or susceptibility. We have performed a detailed study on the expression of IFN-gamma and of the IFN-gamma-inducing cytokines IL-12 and IFN-gamma-inducing factor (IGIF)/IL-18 with regard to time course and tissue localization. IFN-gamma was present in high amounts in the serum and in the supernatants of unseparated spleen cells and isolated CD4+ and CD8+ T cells from the spleens of infected mice which were stimulated ex vivo with T. cruzi. Using the in situ hybridization technique we demonstrate that IL-12 p40 messages were expressed in the spleen and increased during infection, correlating with the expression of IFN-gamma transcripts. Furthermore, we show for the first time that the mRNA for the cytokine IL-18 was induced by a parasitic infection and that this expression increased during infection with T. cruzi. Interestingly, the message for IL-18 was produced earlier during infection and already had declined until day 38, when IFN-gamma and IL-12 p40 transcripts were optimally expressed. Surprisingly, the changes in IL-12 and IL-18 mRNA production were clearly seen only by in situ hybridization, but less clearly by quantitative reverse-transcriptase polymerase chain reaction (RT-PCR). This is possibly due to the extensive activation and proliferation of spleen cells observed during infection leading to a dilution of these specific mRNAs. PMID- 9409640 TI - Impaired Ca2+ mobilization by X-linked agammaglobulinaemia (XLA) B cells in response to ligation of the B cell receptor (BCR). AB - XLA bone marrow samples were shown to contain B cells expressing IgM, and pre-B cells that express the mu-surrogate light chain (mu psiLC) complex, albeit at a reduced frequency to that found in normal bone marrow. Antibody ligation of mu heavy chain on these cells and an XLA B cell line did not induce a Ca2+ flux, whereas ligation of mu heavy chain on normal bone marrow cells, mu psiLC+ pre-B cell lines and an IgM+ B cell line did. The block in XLA B cells was not due to a defect in the basic mechanism of Ca2+ flux generation, as the cells responded well to thapsigargin. In addition, the defect did not affect T cells, which were shown to respond to CD3 antibody with a Ca2+ flux. Ligation of mu heavy chain on XLA bone marrow cells did, however, activate tyrosine kinases, resulting in tyrosine phosphorylation of a cellular protein with a molecular weight of approximately 115 kD. These results indicate that Btk may be necessary for the generation of the Ca2+ flux in response to ligation of mu heavy chain on B cells and mu psiLC+ pre-B. PMID- 9409641 TI - B cells and monocytes are not developmentally affected in a case of reticular dysgenesis. AB - We report a patient with reticular dysgenesis (RD) who received an HLA-identical marrow graft and remained free of infection in spite of incomplete haematological recovery. Mixed chimerism was achieved and resulted from the presence of autologous B cells and monocytes and grafting of donor T cells. Granulocyte recovery was impaired. The B cells were CD5+ (B1 cells) and appeared to be functional, since serum immunoglobulin levels became normal after the graft. The findings described here suggest that in some cases the defect selectively affects different cell types, including the more abundant leucocyte populations, granulocytes and T lymphocytes. However, B cells and monocytes appear to be relatively spared in this case of RD. Furthermore, the present case may provide insight into the mechanism involved in the expansion of distinct B cell subpopulations (B1 and B2 cells). PMID- 9409642 TI - Phenotypic characterization and analysis of allogeneic T cell responses in ZAP-70 dominant negative transgenic mice. AB - Antigen stimulation of T cells results in a series of biochemical events including the interaction of both SH2 domains of ZAP-70 with phosphorylated ITAMS on the T cell receptor. In order to study the physiological relevance of decreasing native ZAP-70-SH2 interaction in vivo, we generated transgenic mice expressing a T cell-specific, dominant negative form of ZAP-70 consisting of only the tandem SH2 domains (ZAP-NC). Phenotypically, these animals had a comparable distribution of lymphocyte subsets in the thymus and spleen compared with the wild-type (WT) controls. However, examination of peripheral blood revealed a slow but progressive decrease in the number of lymphocytes, particularly CD4+ cells, with age (17% reduction by 3 months, 58% reduction by 6 months). Allogeneic responses were then evaluated in vitro as well as in vivo using a subcutaneous sponge matrix implant. Although spleen cells cultured for 4 days in vitro with alloantigen developed normal functional responses, allogeneic responses generated in vivo within a subcutaneous sponge matrix were impaired. This was characterized by a depression in cytotoxic T lymphocyte (CTL) activity, a 82% reduction in the frequency of helper T cells, and a 78% reduction in the capacity of sponge infiltrating lymphocytes to produce IL-2 in response to secondary antigen stimulation. These results indicate that although overt lymphocyte development and in vitro function were unremarkable, expression of a truncated ZAP-70 affected the in vivo survival of peripheral lymphocytes and altered the in vivo generation of functional activity to alloantigen. PMID- 9409643 TI - Protection against anti-glomerular basement membrane (GBM)-mediated nephritis in C3- and C4-deficient mice. AB - Mice rendered completely deficient of the complement components C3 or C4 were used to determine the influence of complement activation in the heterologous phase of the anti-GBM disease model. In wild-type animals the disease is characterized by a neutrophil infiltrate, capillary thrombosis, proteinuria and C3 and C4 deposited within the glomerulus. The early infiltration of neutrophils into the glomeruli is greater in wild-type mice (2.8 +/- 0.3) compared with C3 deficient (1.4 +/- 0.2) and C4-deficient (1.2 +/- 0.003) mice. Deficiency also protects against the subsequent development of proteinuria (2.99 +/- 1.11 mg/24h, 0.059 mg/24h and 0.327 +/- 0.14 mg/24h in wild-type, C3-deficient and C4 deficient mice, respectively) and decreases glomerular capillary thrombosis in both C3- and C4-deficient mice. The degree of protection is greater in the C3 deficient than the C4-deficient animals, suggesting both classical and alternative pathway involvement. These studies support a critical role for complement in the development of anti-GBM disease. However, the protective effect of complement deficiency can be broken if the dose of nephritogenic antibody is increased. PMID- 9409644 TI - Intracellular cytokine expression in whole blood preparations from normals and patients with atopic dermatitis. AB - In recent years, the importance of characterizing the role of cytokines in a wide range of clinical conditions has resulted in development of new methods to assess cytokine expression in clinical samples. The use of anti-cytokine MoAbs and flow cytometry to detect cytokines intracellularly at the single-cell level has the potential to quantify cytokine production in different diseases. For this technique to be useful in a clinical setting, rapid throughput of clinical samples and a cheap, reliable assay would be required, therefore the development of the above technique using unseparated whole blood samples would be advantageous. Using this technique, only one study to date (Maino et al., 1996) has used unseparated whole blood as the source of cells for detecting intracellular cytokines. In clinical practice, whole blood may be optimal, since this most closely approximates conditions in vivo: as no purification of blood mononuclear cells is required, very little blood is needed to detect a number of cytokines simultaneously in various lymphocyte subpopulations, and the assay can be applied to samples from infants and children. In this study we describe an intracellular cytokine assay using unseparated whole blood from normals. In activated CD8- T cells, IL-2 and interferon-gamma (IFN-gamma) were optimally induced after 10 h stimulation with phorbol 12-myristate acetate (PMA)/ionomycin, and in CD8+ T cells IL-2 was optimally induced after 10 h and IFN-gamma after 6 h. The levels of IL-2 and IFN-gamma in CD8+ and CD8- T cells in four healthy individuals were consistent on four occasions over a 3-month period. In a large group of 34 normal subjects, there was considerable heterogeneity in CD3/IL-2+ (range 9.7-41.3) and CD3/IFN-gamma+ cells (10.1-44), expressed as a percentage of total lymphocytes. In patients with atopic dermatitis (n = 5) there was a significantly decreased percentage of CD3+/CD8+ peripheral blood T cells expressing IFN-gamma and an increased percentage of CD3+/CD8- T cells expressing IL-4 compared with non-atopic dermatitis controls (n = 5). Possible applications of this technique are discussed. PMID- 9409645 TI - In vitro effects of mercuric chloride (HgCl2) on human mononuclear cells. AB - Due to the release of the toxic compounds of mercury from amalgam fillings, dental amalgam has been questioned as an adequate restoration material for tooth fillings. HgCl2 has been found to be mitogenic for human blood lymphocytes in vitro. However, activation required much higher concentrations than are ever found in vivo. This study has been initiated to evaluate further the influence of HgCl2 on human immunocompetent cells in vitro. It is found that HgCl2 in a narrow concentration range has the ability to preferentially stimulate the CD4+ T cell subset to blast transformation and DNA synthesis. The reaction, when monitored during days 2-6, is maximal at day 6, and most blasts express the IL-2 receptor (IL-2R), indicating in vitro activation. The CD8+ T cell subset is not affected to the same extent. In addition, HgCl2-induced lymphocyte reactivity is dependent on accessory cells, i.e. CD14+ cells. PMID- 9409646 TI - Autonomic nervous dysfunction in systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA): possible pathogenic role of autoantibodies to autonomic nervous structures. AB - Autonomic nervous dysfunction has been previously reported in SLE, RA and systemic sclerosis, but the pathogenesis of such a complication is poorly understood. In the present study, four standard cardiovascular autonomic function tests were performed in 34 female patients with connective tissue diseases and in 25 healthy control subjects, and results expressed as cardiovascular (CV) test scores. Moreover, in each subject the presence of circulating complement-fixing autoantibodies directed against sympathetic and parasympathetic nervous structures, represented by superior cervical ganglia and vagus nerve, respectively, was simultaneously assessed by an indirect immunofluorescent complement-fixation technique, using rabbit tissue as substrate. None of the patients reported autonomic symptoms. However, an abnormal CV test score (> or = 5) was detected in 15% of the patients and in none of the healthy control subjects, approaching statistical significance (P = 0.07). No correlation was found between CV test results and disease duration, type of therapy or presence of conventional autoantibodies. One or two autoantibodies to autonomic nervous structures were detected in six patients (18%) and not in the control subjects (P < 0.05). Values of deep breathing test were significantly lower in autoantibody positive patients compared with those amongst the control subjects (P < 0.05), and an abnormal CV test score was significantly associated with the presence of autoantibodies to autonomic nervous structures (P < 0.05). In conclusion, we confirm that autonomic nervous function can be impaired in patients with connective tissue diseases, and suggest that autoantibodies directed against autonomic nervous system structures may play a role in the pathogenesis of the autonomic dysfunction. PMID- 9409648 TI - Modulation of Fas-mediated apoptosis of human thyroid epithelial cells by IgG from patients with Graves' disease (GD) and idiopathic myxoedema. AB - The expression of two autoimmune thyroid diseases. GD and idiopathic myxoedema, is associated with antibodies to the thyroid-stimulating hormone (TSH) receptor. Thyroid stimulating antibodies (TSAb) in GD are TSH agonists and cause hyperthyroidism as well as goitre, whereas thyroid stimulation blocking antibodies (TSBAb) in idiopathic myxoedema are TSH antagonists and cause hypothyroidism and thyroid atrophy. We investigated the effect of antibodies to TSH receptor on Fas-mediated apoptosis of thyroid epithelial cells (thyrocytes). Human IgG was isolated from healthy donors, patients with GD and idiopathic myxoedema. Human thyrocytes were obtained from surgical specimens. Thyrocytes were cultured in the presence or absence of human IgG with or without interferon gamma (IFN-gamma) or IL-1beta for a specified time. After incubation, we examined the level of cAMP in cultured supernatants and both Fas and Bcl-2 expression on thyrocytes. In addition, we examined anti-Fas-mediated apoptosis of thyrocytes. Fas expression on thyrocytes was significantly down-regulated by Graves' IgG and TSH, although idiopathic myxoedema IgG did not affect Fas expression on thyrocytes. Idiopathic myxoedema IgG abrogated the effect of TSH on both cAMP production and inhibition of Fas expression on thyrocytes. Treatment of thyrocytes with IL-1beta or IFN-gamma caused a marked augmentation of Fas expression on thyrocytes. The increase of Fas expression of thyrocytes induced by IL-1beta or IFN-gamma was significantly suppressed in the presence of TSH or Graves' IgG. Anti-Fas-induced apoptosis of thyrocytes was observed in thyrocytes treated with IL-1beta or IFN-gamma, but was markedly inhibited in the presence of TSH or Graves' IgG. Furthermore, idiopathic myxoedema IgG abrogated most of the inhibitory effect of TSH on Fas-mediated apoptosis of thyrocytes treated with IL 1beta or IFN-gamma. Bcl-2 expression of thyrocytes did not change after stimulation with TSH, Graves' IgG, idiopathic myxoedema IgG, IL-1beta or IFN gamma. These results suggest that TSAb found in Graves' patients may be potentially involved in the development of goitre by inhibition of Fas-mediated apoptosis of thyrocytes. In addition, TSBAb inhibit the action of TSH and increase the sensitivity toward Fas-mediated apoptosis of thyrocytes, inducing thyroid atrophy seen in patients with idiopathic myxoedema. PMID- 9409647 TI - HLA-dependent peripheral T cell receptor (TCR) repertoire formation and its modification by rheumatoid arthritis (RA) AB - Our previous studies have disclosed that the peripheral T cell receptor beta (TCRB) gene repertoires of RA monozygotic twins were similar. This suggested that the TCRBV repertoire is controlled primarily by genetic factors. Here, we examine how the combination of HLA and presence of RA influence the peripheral TCRB repertoire. Peripheral blood mononuclear cells from six pairs of healthy monozygotic twins, six pairs of monozygotic twins discordant for RA, and nine siblings of a large family, including three RA patients, were examined for their TCRB gene repertoires. Among healthy twins and siblings, the BV repertoires between HLA-identical pairs were significantly more similar than those of HLA-non identical pairs. When RA-affected members were included, the repertoires of the HLA-identical pairs discordant for RA were dissimilar compared with those of healthy pairs. TCRBV-BJ combination repertoire analysis of CD4 and CD8 T cell subsets from the twins showed that the dissimilarity was primarily confined to CD8 T cells in the healthy identical twins, whereas it was seen in both CD4 and CD8 T cell subsets in the RA-discordant twins. These results suggest (i) the presence of RA modifies the genetically controlled TCR repertoire of peripheral T cells, and (ii) the RA-associated alterations appear to occur more frequently in CD4 T cells than in CD8 T cells. PMID- 9409649 TI - Analysis of T cell receptor Vbeta usage in the autoimmune sialadenitis of non obese diabetic (NOD) mice. AB - The NOD mouse develops spontaneous autoimmune sialadenitis besides a well characterized T cell-mediated autoimmune insulitis. We used reverse transcriptase polymerase chain reaction (RT-PCR) to analyse the repertoire of T cell receptor (TCR) Vbeta chain genes expressed in the isolated infiltrating cells from affected salivary glands. Immunohistochemical analysis showed that the vast majority of inflammatory infiltrates in the salivary glands were CD4+ Vbeta8+ and CD4+ Vbeta6+ T cells, whereas CD8+ T cells and B220+ B cells were fewer in number. Predominant expression of the Vbeta8 and Vbeta6 gene segment was detected in the infiltrating cells in the salivary glands very early, and age-related diversity of TCR Vbeta gene usage was observed. Single-strand conformation polymorphism (SSCP) analysis demonstrated a strikingly symmetrical distribution of expanded clones in the PCR products of the Vbeta8 and Vbeta6 gene in the cells infiltrating the salivary glands. Nucleotide sequencing of amplified TCR Vbeta cDNA revealed that T cell clonotypes had a high incidence of identical clones, indicating that the immune response in NOD sialadenitis is driven by common stimuli. These findings suggest that in spontaneous autoimmune sialadenitis of NOD mice, there may be a restricted usage of TCR Vbeta elements in the early stage of the autoimmune lesion to recognize self-antigen in organ-specific autoimmune sialadenitis. PMID- 9409650 TI - The avidity of allospecific cytotoxic T lymphocytes (CTL) determines their cytokine production profile. AB - Donor-specific CTL present within the cardiac allograft during a rejection episode are distinct from those that populate the cardiac allograft in the absence of rejection. Whereas the former generally have a high avidity for donor cells, the latter mainly have a low avidity for donor cells. This observation made us reason that high-avidity CTL are implicated in transplant rejection, whereas low-avidity CTL are not. In the present study, we analyse whether both CTL subsets were distinct with respect to their IL-2, IL-4, IL-6 and interferon gamma (IFN-gamma) secretion pattern. CTL clones with either a high or a low avidity for donor antigens were stimulated with donor cells, third party cells, or immobilized anti-CD3 MoAb and the amount of cytokine released was measured. High- and low-avidity CTL clones were found to differ with respect to their IFN gamma production profile. Stimulation with donor cells resulted in IFN-gamma secretion by high-avidity CTL clones, but not by low-avidity CTL clones. CD3 stimulation, in contrast, led to secretion of equivalent amounts of IFN-gamma by both CTL subsets. These observations indicate that low-avidity CTL are fully capable of producing IFN-gamma, but, in contrast to high avidity CTL, fail to do so when they encounter donor cells. As IFN-gamma favours the occurrence of transplant rejection, this observation emphasizes the relevance of high-avidity CTL in the rejection process. Additionally, the data show that the cytokine production profile of CTL depends on the nature of the stimulus. PMID- 9409651 TI - Antigen-presenting cells (APC) of mice with chronic graft-versus-host disease (GVHD) cause excessive activation-induced death of T helper cells. AB - Both experimental and clinical forms of chronic GVHD have unique immunological features. The affected animals/individuals suffer from autoimmune disorders such as systemic lupus erythematosus (SLE), and yet they are unable to mount a self MHC-restricted T cell response to foreign antigens. Pathogenesis of the latter phenomenon was investigated in an experimental model of chronic GVHD. Chronic GVHD was induced in 8-10-week-old (B6xC3H)F1 mice by tail vein injection of 5 x 10(7) spleen cells of C3H parental strain. The recipients, when tested 3 months later, were unable to mount a T helper (Th) cell response to a randomly selected immunogen, a vaccine of l0(8) killed Mycobacterium vaccae. The animals showed evidence of generalized lymphoid hyperplasia, as indicated by GVH index >1.34, and also revealed autoantibodies against erythrocytes and dsDNA, indicating establishment of chronic GVHD. However, mice with chronic GVHD of only 3 weeks duration were able to mount the Th cell response to M. vaccae. Three consecutive immunizations of these mice at 1-week intervals, with the same immunogen, resulted in the mice becoming non-responsive to the antigen. All the three responses tested, namely the DTH, lymphoproliferation and the antibody responses, were adversely affected. The non-responsiveness induced was antigen-specific. Mice receiving two immunizations with M. vaccae responded normally to Salmonella enteritidis. Pulse treatment with cyclosporin A 0.5 mg/mouse by the i.p. route, on days 0, 1, 2, 3 and 4 at the time of immunization with M. vaccae on day 1, prevented emergence of non-responsiveness. Based on this evidence, it was concluded that repeated activation of T cells of mice with chronic GVHD induces non-responsiveness. Extent of clonal loss due to activation-induced cell death (AICD) caused by i.p. injection with a superantigen Staphylococcal enterotoxin B (SEB) was investigated in F1 mice with chronic GVHD. I.p. injection of 25 microg/mouse of SEB induced loss of SEB responding clones in both normal F1 mice and those having chronic GVHD; however, the extent of loss was much greater in the latter. In vitro antigen-specific proliferation of primed splenic T cells of normal F1 mice was observed to be quite poor when antigen was presented by APC of mice with chronic GVHD of 3 weeks duration. Proliferation profiles of T cells of normal F1 mice, in response to stimulation with concanavalin A (Con A) or SEB, were studied, using as APC irradiated spleen cells of normal F1 mice or of F1 mice with chronic GVHD of 3 weeks duration. With Con A and APC of normal F1 mice, peak proliferation was observed at 48 h, which remained at the same level up to 72 h and declined thereafter, possibly due to AICD. With SEB and the normal APC, proliferation progressively peaked at 72 h and declined thereafter. With APC of mice with chronic GVHD, the 48 h proliferative responses of both Con A and SEB were comparable to those caused by APC of normal F1 mice; however, thereafter the responses declined steeply, suggesting greater AICD. Based on these results, it was concluded that APC of mice with chronic GVHD are functionally altered to induce greater AICD. PMID- 9409652 TI - Analysis of V(H) genes rearranged by individual B cells in dermal infiltrates of patients with mycosis fungoides. AB - In patients with cutaneous T cell lymphomas such as mycosis fungoides B cells can frequently be detected in the lymphocytic dermal infiltrate. To analyse their immunoglobulin heavy chain gene repertoire, single B cells were obtained from tissue sections of two typical patients with mycosis fungoides using hydraulic micromanipulation followed by specific amplification of the respective gene segments by single-cell polymerase chain reaction (PCR) technique. A total of 21 V(H)DJ(H) genes was sequenced. From each individual B cell a single productive V(H)DJ(H) rearrangement was obtained. There was no clonal relationship detected between any of these rearrangements suggesting polyclonality of the infiltrating B cells. The representation of V(H) families was in accordance with the germ-line complexity. A remarkably high number of V(H) genes (5/13 in patient 1; 3/8 in patient 2) was completely or nearly germ-line-identical. Five of seven V(H)4 family genes were nearly unmutated. On the other hand, most of the V(H)3 gene family members were somatically mutated in an antigen-driven manner. The proportion of germ-line-identical V(H) genes, the usage of individual V(H), D, J(H) gene elements, and the pattern of somatic mutations found in the B cells infiltrating skin lesions of patients with mycosis fungoides resembles the peripheral blood repertoire, suggesting a bystander role of these cells. PMID- 9409653 TI - Amino-terminal identity of co-existent amyloid and non-amyloid immunoglobulin kappa light chain deposits. A human disease to study alterations of protein conformation. AB - Tissue deposition of monoclonal immunoglobulin light chains is a serious complication in some patients with B cell proliferative disorders. The deposits are typically fibrillar and Congophilic in amyloid (AL) and non-fibrillar and Congophobic in light chain deposition disease (LCDD), and rarely coexist in the same patient. From post-mortem tissue of an individual with fibrillar and non fibrillar kappa light chain deposits in different sites, we separately extracted and analysed biochemically and immunochemically the non-amyloid deposits from isolated glomeruli, the amyloid from isolated renal arteries and the amyloid from myocardium in which the only deposits were amyloid restricted to mural arteries. Western blotting analysis of both the extracted amyloid and the non-amyloid deposits demonstrated 25-kD bands immunoreactive with anti-kappa antibody, and the identity of the N-terminal amino acid sequences that belong to the variable region kappaIV light chain subgroup. This is the first human disease in which antigenically similar but morphologically different deposits have been separately biochemically analysed. We propose that combined LCDD and AL is an ideal human disease to study the relationships and the factors that influence the conversion of non-amyloidogenic to amyloidogenic conformations. PMID- 9409654 TI - Protection from concanavalin A (Con A)-induced T cell-dependent hepatic lesions and modulation of cytokine release in mice by sodium fusidate. AB - The immunomodulatory effects of the antibiotic sodium fusidate (SF) were tested in a model of T cell-dependent hepatic injury that can be induced in normal mice by a single i.v. injection of Con A. Signs of hepatitis with elevated transaminase activities in plasma, severe infiltration of the liver by neutrophil granulocytes, lymphocytes and monocytes, and necrotic areas were observed in control mice treated intraperitoneally with PBS 24 h and 1 h before Con A challenge. T cell- and macrophage-derived cytokines (IL-2, interferon-gamma (IFN gamma), tumour necrosis factor-alpha (TNF-alpha, IL-1beta, IL-6) were released with different kinetics in the circulation of these mice. SF, 20, 40 or 80 mg/kg, administered 24 h and 1 h before Con A challenge, protected the mice against the hepatitic effects of Con A. The protective effects of SF were dose-dependent and accompanied by profound modifications of blood levels of cytokines induced by Con A, so that, relative to control mice, SF (80 mg/kg)-treated animals showed markedly diminished plasma levels of IL-2, IFN-gamma and TNF-alpha, along with augmented levels of IL-6. These results suggest that SF might be useful in the treatment of immunoinflammatory liver diseases in humans. PMID- 9409655 TI - Inhibitory effects of low molecular weight heparin on mediator release by mast cells: preferential inhibition of cytokine production and mast cell-dependent cutaneous inflammation. AB - There has been substantial evidence that suggests that heparin may modulate various aspects of immune function and inflammation in addition to its well known anticoagulant activity. In this regard heparin was found to suppress cell mediated immune responses or asthmatic reactions to allergen challenge. In the present study we analyse the effects of low molecular weight heparin (LMWH) on mast cell degranulation and cytokine production in vitro and on the elicitation of IgE-mediated mast cell-dependent late cutaneous allergic inflammation in vivo. We have established that LMWH preferentially inhibited tumour necrosis factor alpha (TNF-alpha) and IL-4 production without having any significant effect on mast cell degranulation. These effects have been observed in mast cells derived from three different origins that were activated by either immunological or non immunological stimuli. We have shown that there is inhibition of TNF-alpha production (and not neutralization of activity), as elimination of the drug after a short preincubation and addition of LMWH to rTNF-alpha had no effect on TNF alpha-mediated cytotoxic activity. These results were also confirmed by ELISA. In vivo, s.c. injection of the LMWH inhibited the leucocyte infiltration associated with the late cutaneous response which followed passive cutaneous anaphylaxis (PCA) reaction, without affecting mast cell numbers or degranulation. These data suggest that LMWH may have an inhibitory role in mast cell-mediated allergic inflammation, and thus might be considered as a possible therapeutic modality. PMID- 9409656 TI - Intrathecal release of pro- and anti-inflammatory cytokines during stroke. AB - A growing body of evidence points out the potential role of inflammatory mechanisms in the pathophysiology of ischaemic brain damage. We have recently demonstrated that stroke patients display an intrathecal production of proinflammatory cytokines, such as IL-1beta and IL-6 already within the first 24 h after the beginning of symptoms (Tarkowski et al., 1995). The aim of the present study was to investigate patterns of local inflammatory responses as a consequence of acute stroke. Thirty stroke patients were studied prospectively on days 0-3, 7-9, 21-26 and after day 90 with clinical evaluations, radiological assessments and analysis of cerebrospinal fluid (CSF) cytokine levels. In addition, 15 healthy control CSF samples were used. Significantly increased CSF levels of IL-8, granulocyte-macrophage colony-stimulating factor (GM-CSF) and IL 10 were observed early during the stroke with a peak on day 2 for the proinflammatory cytokines IL-8 and GM-CSF, and on day 3 for the immunoregulatory cytokine IL-10. Patients with a brain infarct predominantly located in the white matter showed significantly higher levels of IL-8 in CSF than patients with an infarct mainly located in the grey matter. Also, high levels of intrathecal tumour necrosis factor-alpha (TNF-alpha) were associated with the presence of white matter disease. Our study demonstrates an intrathecal production of proinflammatory and immunoregulatory cytokines in patients with stroke, supporting the notion of localized immune response to the acute brain lesion. A better understanding of the inflammatory response in stroke may lead to new treatment strategies. PMID- 9409657 TI - Circadian rhythm of leucocytes and lymphocytes subsets and its possible correlation with the function of the autonomic nervous system. AB - There are physiological variations in the levels of leucocytes. Among these, the circadian rhythm is very important in terms of the magnitude. Since newly identified lymphocyte subsets (i.e. extrathymic T cells) have recently been detected, a comprehensive study of the circadian rhythm was conducted. All leucocytes were found to vary in number or proportion with a circadian rhythm and were classified into two groups. One group--granulocytes, macrophages, natural killer (NK) cells, extrathymic T cells, gammadelta T cells, and CD8+ subset- showed an increase in the daytime (i.e. daytime rhythm). The other group--T cells, B cells, alphabeta T cells, and CD4+ subset--showed an increase at night. Humans are active and show sympathetic nerve dominance in the daytime. Interestingly, granulocytes and lymphocyte subsets with the daytime rhythm were found to carry a high density of adrenergic receptors. On the other hand, lymphocyte subsets with the night rhythm carried a high proportion of cholinergic receptors. Reflecting this situation, exercise prominently increased the number of cells with the daytime rhythm. These results suggest that the levels of leucocytes may be under the regulation of the autonomic nervous system. PMID- 9409658 TI - Cross-linking of antigen receptor via Ig-beta (B29, CD79b) can induce both positive and negative signals in CD40-activated human B cells. AB - Antigen-dependent activation of B lymphocytes is mediated through surface immunoglobulins and their associated molecules Ig-alpha (CD79a, Mb1) and Ig-beta (CD79b, B29). Here we show that an antibody directed against the extracellular part of human Ig-beta can, when cross-linked by CD32-transfected L cells, induce an IL-2-dependent proliferation of tonsil B cells. With the use of L cells stably transfected with both CD32 and CD40L, anti-Ig-beta activation of B cells was combined with CD40 triggering, an important component of the T cell-dependent B cell activation. This dual cellular activation resulted in two different phases, with initially synergistic proliferative effects, both without and with IL-2 or IL-10. Then, after 5-6 days of culture, cells stimulated with both anti-Ig-beta and CD40L underwent massive cell death, in contrast to B cells activated with CD40L alone. Cell death was not prevented by the addition of IL-2 or IL-10, but was prevented by the addition of IL-4. These results are discussed in the context of positive and negative selection of mature B cells. PMID- 9409659 TI - N-3 polyunsaturated fatty acids modulate the expression of functionally associated molecules on human monocytes and inhibit antigen-presentation in vitro. AB - N-3 polyunsaturated fatty acid (PUFA)-rich diets are associated with suppression of cell-mediated immune responses, but the mechanisms are unclear. Specific immune responses are initiated by antigen-presenting cells (APC). We have previously shown in vitro that the n-3 PUFA, eicosapentaenoic acid (EPA), inhibits the expression of HLA-DR, an MHC class II molecule required for normal APC function on human blood monocytes. In contrast, docosahexaenoic acid (DHA) enhanced the expression of this molecule on unstimulated monocytes, but both n-3 PUFA suppressed its expression on interferon-gamma (IFN-gamma)-activated monocytes. In the present study we show that when EPA and DHA were combined at the same ratio as is commonly found in fish oil supplement capsules (3:2) there was no significant effect in vitro on the expression of HLA-DR on unstimulated monocytes, but the expression on IFN-gamma-activated monocytes remained significantly inhibited. In the same in vitro system a significant reduction in the ability of IFN-gamma-activated monocytes to present tetanus toxoid antigen to autologous lymphocytes was observed following culture with the combined n-3 PUFA. These findings support previous animal studies which suggest that n-3 PUFA can inhibit the antigen-presenting function of mononuclear phagocytes. PMID- 9409660 TI - Human peripheral eosinophils express functional interferon-gamma receptors (IFN gammaR). AB - In order to determine whether or not IFN-gammaR is associated with regulatory mechanisms on human eosinophil function, we examined the expression of functional IFN-gammaR on human peripheral eosinophils. In this study, peripheral blood eosinophils were obtained from seven normal controls and 12 patients (bronchial asthma, n = 9, and hypereosinophilic syndrome (HES), n = 3), and the purity of eosinophils was 97.11 +/- 2.31%, n = 19. We first showed that anti-IFN-gammaR alpha-chain MoAb reacted with all tested eosinophils of both normal controls and patients by flow cytometry analysis. We also showed expression of mRNA for the alpha-chain of IFN-gammaR in all purified eosinophils of six individuals. Further, to characterize IFN-gammaR on eosinophils, we did binding experiments with 125I-IFN-gamma on purified peripheral eosinophils. The linear Scatchard plot indicated a single type of high-affinity binding sites (dissociation constant (Kd) = 3.89-4.95 x 10(-10) M, numbers of binding sites = 183-233/cell, n = 3). To determine whether IFN-gammaR on eosinophils is functional, we examined surface eosinophilic cationic protein (ECP) and CD69 induction after IFN-gammaR ligation with recombinant human IFN-gamma (rhIFN-gamma) on eosinophils by flow cytometry. rhIFN-gamma stimulation significantly induced both ECP and CD69 expression on the 2-18 h-cultured eosinophils in a dose-dependent manner. Further, the effects of rhIFN-gamma stimulation were significantly blocked by both a neutralizing anti IFN-gamma MoAb and a blocking anti-IFN-gammaR MoAb. These results suggest that human peripheral eosinophils express functional IFN-gammaR. PMID- 9409661 TI - Regulation of segmentation and segmental identity by Drosophila homeoproteins: the role of DNA binding in functional activity and specificity. AB - Recent advances have shed new light on how the Q50 homeoproteins act in Drosophila. These transcription factors have remarkably similar and promiscuous DNA-binding specificities in vitro; yet they each specify distinct developmental fates in vivo. One current model suggests that, because the Q50 homeoproteins have distinct biological functions, they must each regulate different target genes. According to this 'co-selective binding' model, significant binding of Q50 homeoproteins to functional DNA elements in vivo would be dependent upon cooperative interactions with other transcription factors (cofactors). If the Q50 homeoproteins each interact differently with cofactors, they could be selectively targeted to unique, limited subsets of their in vitro recognition sites and thus control different genes. However, a variety of experiments question this model. Molecular and genetic experiments suggest that the Q50 homeoproteins do not regulate very distinct sets of genes. Instead, they mostly control the expression of a large number of shared targets. The distinct morphogenic properties of the various Q50 homeoproteins may principally result from the different manners in which they either activate or repress these common targets. Further, in vivo binding studies indicate that at least two Q50 homeoproteins have very broad and similar DNA-binding specificities in embryos, a result that is inconsistent with the 'co-selective binding' model. Based on these and other data, we suggest that Q50 homeoproteins bind many of their recognition sites without the aid of cofactors. In this 'widespread binding' model, cofactors act mainly by helping to distinguish the way in which homeoproteins regulate targets to which they are already bound. PMID- 9409662 TI - Only a subset of the binary cell fate decisions mediated by Numb/Notch signaling in Drosophila sensory organ lineage requires Suppressor of Hairless. AB - In Drosophila, an adult external sensory organ (bristle) consists of four distinct cells which arise from a sensory organ precursor cell via two rounds of asymmetric divisions. The sensory organ precursor cell first divides to generate two secondary precursor cells, IIa and IIb. The IIa cell then divides to produce the hair cell and the socket cell. Shortly after, the IIb cell divides to generate the neuron and the sheath cell. The membrane-associated protein Numb has been shown to be required for the first two asymmetric divisions. We now report that a new hypomorphic numb mutant not only displays a double-socket phenotype, due to a hair cell to socket cell transformation, but also a double-sheath phenotype, due to a neuron to sheath cell transformation. This provides direct evidence that numb functions in the neuron/sheath cell lineage as well. Those results, together with our observation from immunofluorescence analysis that Numb forms a crescent in the dividing IIa and IIb cells suggest that asymmetric localization of Numb is important for the cell fate determination in all three asymmetric cell divisions in the sensory organ lineage. Interestingly, we found that in the hair/socket cell lineage but not the neuron/sheath cell lineage, a Suppressor of Hairless mutation acts as a dominant suppressor of numb mutations whereas Hairless mutations act as enhancers of numb. Moreover, epistasis analysis indicates that Suppressor of Hairless acts downstream of numb, and results from in vitro binding analysis suggest that the genetic interaction between numb and Hairless may occur through direct protein-protein interaction. These studies reveal that Suppressor of Hairless is required for only a subset of the asymmetric divisions that depend on the function of numb and Notch. PMID- 9409663 TI - Fkh5-deficient mice show dysgenesis in the caudal midbrain and hypothalamic mammillary body. AB - The murine winged helix gene Fkh5 is specifically expressed in the developing central nervous system (CNS). Early embryonic Fkh5 expression is restricted to the mammiliary body region of the caudal hypothalamus, midbrain, hindbrain and spinal cord. Postnatally, signals persist in specific nuclei of the mammillary body and in the midbrain. We generated Fkh5 deficient mice by homologous recombination to assess its in vivo function. At birth, Fkh5-deficient mice are viable and indistinguishable from wild-type and Fkh5 heterozygous littermates. However, about one third die within the first two days and another fifth before weaning. Surviving Fkh5-deficient mice become growth retarded within the first week and remain smaller throughout their whole life span. Fkh5-deficient females on 129Sv x C57BL/6 genetic background are fertile, but do not nurture their pups. More detailed analysis of Fkh5-deficient brains reveals distinct alterations in the CNS. In the midbrain, mutant mice exhibit reduced inferior colliculi and an overgrown anterior cerebellum. Furthermore, the hypothalamic mammillary body of Fkh5-deficient brains lacks the medial mammillary nucleus. These results suggest that Fkh5 plays a major role during CNS development. PMID- 9409664 TI - The molecular nature of zebrafish swirl: BMP2 function is essential during early dorsoventral patterning. AB - Early dorsoventral pattern formation in vertebrate embryos is regulated by opposing activities of ventralizing bone morphogenetic proteins (BMPs) and dorsal specific BMP antagonists such as Chordin, Noggin and Follistatin. Specific defects in early dorsoventral patterning have been recently found in a number of zebrafish mutants, which exhibit either a ventralized or dorsalized phenotype. One of these, the ventralized mutant chordino (originally called dino) is caused by a mutation in the zebrafish chordin homologue and interacts genetically with the dorsalized mutant swirl. In swirl mutant embryos, dorsal structures such as notochord and somites are expanded while ventral structures such as blood and nephros are missing. Here we demonstrate that the swirl phenotype is caused by mutations in the zebrafish bmp2 gene (zbmp2). While injection of mRNAs encoded by the mutant alleles has no ventralizing effect, injection of wild-type zbmp2 mRNA leads to a complete rescue of the swirl mutant phenotype. Fertile adult mutant fish were obtained, showing that development after gastrulation is not dependent on zbmp2 function. In addition zBMP2 has no maternal role in mesoderm induction. Our analysis shows that swirl/BMP2, unlike mouse BMP2 but like mouse BMP4, is required for early dorsoventral patterning of the zebrafish embryo. PMID- 9409665 TI - Cellular interpretation of multiple TGF-beta signals: intracellular antagonism between activin/BVg1 and BMP-2/4 signaling mediated by Smads. AB - During early embryogenesis of Xenopus, dorsoventral polarity of the mesoderm is established by dorsalizing and ventralizing agents, which are presumably mediated by the activity of an activin/BVg1-like protein and Bone Morphogenetic Proteins (BMP), respectively. Interestingly, these two TGF-beta subfamilies are found in overlapping regions during mesoderm patterning. This raises the question of how the presumptive mesodermal cells recognize the multiple TGF-beta signals and differentially interpret this information to assign a particular cell fate. In this study, we have exploited the well characterized model of Xenopus mesoderm induction to determine the intracellular interactions between BMP-2/4 and activin/BVg1 signaling cascades. Using a constitutively active BMP-2/4 receptor that transduces BMP-2/4 signals in a ligand-independent fashion, we demonstrate that signals provided by activin/BVg1 and BMP modulate each other's activity and that this crosstalk occurs through intracellular mechanisms. In assays using BMP 2/4 and activin/BVg1-specific reporters, we determined that the specificity of BMP-2/4 and activin/BVg1 signaling is mediated by Smad1 and Smad2, respectively. These Smads should be considered as the mediators of the intracellular antagonism between BMP-2/4 and activin/BVg1 signaling possibly through sequestration of a limited pool of Smad4. Consistent with such a mechanism, Smad4 interacts functionally with both Smad1 and -2 to potentiate their signaling activities, and a dominant negative variant of Smad4 can inhibit both activin/BVg1 and BMP-2/4 mediated signaling Finally, we demonstrate that an activin/BVg1-dependent transcriptional complex contains both Smad2 and Smad4 and thereby provides a physical basis for the functional involvement of both Smads in TGF-beta-dependent transcriptional regulation. Thus, Smad4 plays a central role in synergistically activating activin/BVg1 and BMP-dependent transcription and functions as an intracellular sensor for TGF-beta-related signals. PMID- 9409666 TI - ETTIN patterns the Arabidopsis floral meristem and reproductive organs. AB - ettin (ett) mutations have pleiotropic effects on Arabidopsis flower development, causing increases in perianth organ number, decreases in stamen number and anther formation, and apical-basal patterning defects in the gynoecium. The ETTIN gene was cloned and encodes a protein with homology to DNA binding proteins which bind to auxin response elements. ETT transcript is expressed throughout stage 1 floral meristems and subsequently resolves to a complex pattern within petal, stamen and carpel primordia. The data suggest that ETT functions to impart regional identity in floral meristems that affects perianth organ number spacing, stamen formation, and regional differentiation in stamens and the gynoecium. During stage 5, ETT expression appears in a ring at the top of the floral meristem before morphological appearance of the gynoecium, consistent with the proposal that ETT is involved in prepatterning apical and basal boundaries in the gynoecium primordium. Double mutant analyses and expression studies show that although ETT transcriptional activation occurs independently of the meristem and organ identity genes LEAFY, APETELA1, APETELA2 and AGAMOUS, the functioning of these genes is necessary for ETT activity. Double mutant analyses also demonstrate that ETT functions independently of the 'b' class genes APETELA3 and PISTILLATA. Lastly, double mutant analyses suggest that ETT control of floral organ number acts independently of CLAVATA loci and redundantly with PERIANTHIA. PMID- 9409667 TI - PAX2 is expressed in multiple spinal cord interneurons, including a population of EN1+ interneurons that require PAX6 for their development. AB - Members of the PAX family of transcription factors are candidates for controlling cell identity in the spinal cord. We have morphologically analyzed cells that express one of these transcription factors, PAX2, demonstrating multiple interneuron cell types express PAX2. Two ventral populations of PAX2-expressing interneurons in the spinal cord are marked by coexpression of the transcription factors, EN1 and EVX1. Interestingly, the expression domains of PAX2, EN1 and EVX1 in postmitotic neurons correlate closely with those of Pax6 and Pax7 in the ventricular zone, implicating these patterning genes in the regulation of PAX2, EN1 and EVX1. We show that one of these patterning genes, Pax6, is required for the correct specification of ventral PAX2+ interneurons that coexpress EN1. These results demonstrate that the early activity of patterning genes in the ventricular zone determines interneuron identity in the spinal cord. PMID- 9409668 TI - Targeted disruption of Hoxd-10 affects mouse hindlimb development. AB - Targeted disruption of the Hoxd-10 gene, a 5' member of the mouse HoxD linkage group, produces mice with hindlimb-specific defects in gait and adduction. To determine the underlying causes of this locomotor defect, mutant mice were examined for skeletal, muscular and neural abnormalities. Mutant mice exhibit alterations in the vertebral column and in the bones of the hindlimb. Sacral vertebrae beginning at the level of S2 exhibit homeotic transformations to adopt the morphology of the next most anterior vertebra. In the hindlimb, there is an anterior shift in the position of the patella, an occasional production of an anterior sesamoid bone, and an outward rotation of the lower part of the leg, all of which contribute to the defects in locomotion. No major alterations in hindlimb musculature were observed, but defects in the nervous system were evident. There was a decrease in the number of spinal segments projecting nerve fibers through the sacral plexus to innervate the musculature of the hindlimb. Deletion of a hindlimb nerve was seen in some animals, and a shift was evident in the position of the lumbar lateral motor column. These observations suggest a role for the Hoxd-10 gene in establishing regional identity within the spinal cord and imply that patterning of the spinal cord may have intrinsic components and is not completely imposed by the surrounding mesoderm. PMID- 9409669 TI - Linking Frizzled and Wnt signaling in Drosophila development. AB - Drosophila Frizzled-2 (Dfz2) has been identified as a putative fly Wingless (Wg) receptor. Although Dfz2 shows significant homology with Fz, a protein that operates in the mechanisms that establish planar polarity in Drosophila epithelia, any clear evidence for an involvement by Fz in a Wnt signaling pathway has hitherto been absent. Here we describe the planar polarity phenotypes of loss of-function and overexpression of Fz in the developing Drosophila eye and find it almost identical to the loss-of-function or overexpression of Dishevelled (Dsh - a protein operating in Wnt second messenger systems). In addition, we show that overexpression of Shaggy (Sgg - another component of the Wnt pathway) in the eye also causes a phenotype similar to Fz and Dsh. To test further the link between planar polarity and Wnt signaling we misexpressed Wg in the developing eye and found it had a potent polarizing effect in the retinal epithelium. Since the overexpression of Fz in the developing eye gave a phenotype consistent with activating the Wnt pathway, we tested overexpression of Fz in the developing embryonic ectoderm and found that it phenocopied overexpression of Wg. To check that Fz was indeed able to activate a Wnt pathway we overexpressed it in Drosophila tissue culture cells and observed the characteristic phosphorylation of Dsh that occurs in response to Wnt signaling. Taken together our results significantly strengthen the case for Fz acting in a Wnt signaling pathway in Drosophila. PMID- 9409670 TI - Hoxd-12 differentially affects preaxial and postaxial chondrogenic branches in the limb and regulates Sonic hedgehog in a positive feedback loop. AB - Several 5' members of the Hoxd cluster are expressed in nested posterior-distal domains of the limb bud suggesting a role in regulating anteroposterior pattern of skeletal elements. While loss-of-function mutants have demonstrated a regulatory role for these genes in the developing limb, extensive functional overlaps between various different Hox genes has hampered elucidation of the roles played by individual members. In particular, the function of Hoxd-12 in the limb remains obscure. Using a gain-of-function approach, we find that Hoxd-12 misexpression in transgenic mice produces apparent transformations of anterior digits to posterior morphology and digit duplications, while associated tibial hemimelia and other changes indicate that formation/growth of certain skeletal elements is selectively inhibited. If the digital arch represents an anterior bending of the main limb axis, then the results are all reconcilable with a model in which Hoxd-12 promotes formation of postaxial chondrogenic condensations branching from this main axis (including the anteriormost digit) and selectively antagonizes formation of 'true' preaxial condensations that branch from this main axis (such as the tibia). Hoxd-12 misexpression can also induce ectopic Sonic hedgehog (Shh) expression, resulting in mirror-image polydactyly in the limb. Misexpression of Hoxd-12 in other lateral plate derivatives (sternum, pelvis) likewise phenocopies several luxoid/luxate class mouse mutants that all share ectopic Shh signalling. This suggests that feedback activation of Shh expression may be a major function of Hoxd-12. Hoxd-12 can bind to and transactivate the Shh promoter in vitro. Furthermore, expression of either exogenous Hoxd-11 or Hoxd-12 in cultured limb bud cells, together with FGF, induces expression of the endogenous Shh gene. Together these results suggest that certain 5' Hoxd genes directly amplify the posterior Shh polarizing signal in a reinforcing positive feedback loop during limb bud outgrowth. PMID- 9409671 TI - Pdd1p associates with germline-restricted chromatin and a second novel anlagen enriched protein in developmentally programmed DNA elimination structures. AB - Programmed DNA rearrangements, including DNA diminution, characterize the differentiation of somatic from germline nuclei in several developmental systems. Pdd1p (Programmed DNA degradation protein 1), a development-restricted polypeptide, has been implicated in heterochromatin assembly and DNA degradation during ciliate macronuclear development. Here, cross-linking and co immunoprecipitation were used to verify that Pdd1p-associated chromatin is enriched in germline-restricted DNA. Pdd1p-associated proteins include general core histones and a second anlagen-enriched polypeptide (Pdd2p, formerly known as p43). Immunoblotting analyses demonstrate that, like Pdd1p, Pdd2p is developmentally regulated and present in conjugating cells during the time of germline DNA rearrangements and degradation. Pdd2p is post-translationally modified by phosphorylation at a time in development corresponding to dephosphorylation of Pdd1p and the formation of heterochromatic DNA elimination structures. Following gene cloning, the derived amino acid sequence of the PDD2 gene predicts a novel polypeptide containing multiple putative phosphorylation sites. In situ analyses, using both light and electron microscopy, demonstrate that Pdd1p and Pdd2p co-localize in DNA elimination structures within developing macronuclei. However, unlike Pdd1p, which also localizes to apoptotic macronuclei, Pdd2p appears to be restricted to a higher degree to germline DNA elimination structures. Taken together, the data presented here demonstrate a physical link between Pdd1p and germline-restricted chromatin and establish Pdd2p as the second member of a small group of developmentally restricted polypeptides implicated in programmed DNA elimination. PMID- 9409672 TI - Dorso-ventral ectodermal compartments and origin of apical ectodermal ridge in developing chick limb. AB - We wish to understand how limbs are positioned with respect to the dorso-ventral axis of the body in vertebrate embryos, and how different regions of limb bud ectoderm, i.e. dorsal ectoderm, apical ridge and ventral ectoderm, originate. Signals from dorsal and ventral ectoderm control dorso-ventral patterning while the apical ectodermal ridge (AER) controls bud outgrowth and patterning along the proximo-distal axis. We show, using cell-fate tracers, the existence of two distinct ectodermal compartments, dorsal versus ventral, in both presumptive limb and flank of early chick embryos. This organisation of limb ectoderm is the first direct evidence, in vertebrates, of compartments in non-neural ectoderm. Since the apical ridge appears to be confined to this compartment boundary, this positions the limb. The mesoderm, unlike the ectoderm, does not contain two separate dorsal and ventral cell lineages, suggesting that dorsal and ventral ectoderm compartments may be important to ensure appropriate control of mesodermal cell fate. Surprisingly, we also show that cells which form the apical ridge are initially scattered in a wide region of early ectoderm and that both dorsal and ventral ectoderm cells contribute to the apical ridge, intermingling to some extent within it. PMID- 9409673 TI - The activity of neurogenin1 is controlled by local cues in the zebrafish embryo. AB - Zebrafish neurogenin1 encodes a basic helix-loop-helix protein which shares structural and functional characteristics with proneural genes of Drosophila melanogaster. neurogenin1 is expressed in the early neural plate in domains comprising more cells than the primary neurons known to develop from these regions and its expression is modulated by Delta/Notch signalling, suggesting that it is a target of lateral inhibition. Misexpression of neurogenin1 in the embryo results in development of ectopic neurons. Markers for different neuronal subtypes are not ectopically expressed in the same patterns in neurogenin1 injected embryos suggesting that the final identity of the ectopically induced neurons is modulated by local cues. Induction of ectopic motor neurons by neurogeninl requires coexpression of a dominant negative regulatory subunit of protein kinase A, an intracellular transducer of hedgehog signals. Moreover, the pattern of endogenous neurogenin1 expression in the neural plate is expanded in response to elevated levels of Hedgehog (Hh) signalling or abolished as a result of inhibition of Hh signalling. Together these data suggest that Hh signals regulate neurogenin1 expression and subsequently modulate the type of neurons produced by Neurogenin1 activity. PMID- 9409674 TI - The Drosophila tango gene encodes a bHLH-PAS protein that is orthologous to mammalian Arnt and controls CNS midline and tracheal development. AB - The Drosophila single-minded and trachealess bHLH-PAS genes control transcription and development of the CNS midline cell lineage and tracheal tubules, respectively. We show that Single-minded and Trachealess activate transcription by forming dimers with the Drosophila Tango protein that is an orthologue of the mammalian Arnt protein. Both cell culture and in vivo studies show that a DNA enhancer element acts as a binding site for both Single-minded::Tango and Trachealess::Tango heterodimers and functions in controlling CNS midline and tracheal transcription. Isolation and analysis of tango mutants reveal CNS midline and tracheal defects, and gene dosage studies demonstrate in vivo interactions between single-minded::tango and trachealess::tango. These experiments support the existence of an evolutionarily conserved, functionally diverse bHLH-PAS protein regulatory system. PMID- 9409675 TI - A novel alpha integrin subunit associates with betaPS and functions in tissue morphogenesis and movement during Drosophila development. AB - We have identified a novel alpha integrin subunit in Drosophila, that associates with betaPS integrin. We report the temporal expression of the gene encoding this integrin subunit, which we have called alphaPS3, throughout development and the localization of its expression during embryogenesis. AlphaPS3 RNA was localized to tissues undergoing invagination, tissue movement and morphogenesis such as salivary gland, trachea, midgut, dorsal vessel, midline of the ventral nerve cord, amnioserosa and the amnioproctodeal invagination. AlphaPS3 DNA localized to the chromosomal vicinity of scab (scb), previously identified by a failure of dorsal closure. Embryos homozygous for the 119 allele of scb had no detectable alphaPS3 RNA and the 1035 allele of scb contains a P element inserted just 5' of the coding region for the shorter of the gene's two transcripts. Furthermore, mutations in the scb locus exhibit additional defects corresponding to sites of alphaPS3 transcription, including abnormal salivary glands, mislocalization of the pericardial cells and interrupted trachea. Removal of both maternal and zygotic betaPS produced similar defects, indicating that these two integrin subunits associate in vivo and function in the movement and morphogenesis of tissues during development in Drosophila. Phenotypic similarities suggest that laminin A is a potential ligand for this integrin, at least in some tissues. PMID- 9409676 TI - HSP70-2 is required for desynapsis of synaptonemal complexes during meiotic prophase in juvenile and adult mouse spermatocytes. AB - Spermatogenic cells synthesize a unique 70-kDa heat shock protein (HSP70-2) during prophase of meiosis I, and targeted disruption of the Hsp70-2 gene has shown that this protein is required for spermatogenic cell differentiation in adult mice. HSP70-2 is associated with synaptonemal complexes formed between paired homologous chromosomes during meiotic prophase. The present study focuses on the nearly synchronous first wave of spermatogenesis in 12- to 28-day old juvenile mice to determine more precisely when HSP70-2 is required and what meiotic processes are affected by its absence. Spermatogenesis in homozygous mutant mice (Hsp70-2[-/-]) proceeded normally until day 15 when increasing numbers of pachytene spermatocytes became apoptotic and differentiation of cells beyond the pachytene stage began to falter. Synaptonemal complexes assembled in Hsp70-2(-/-) mice and spermatocytes developed through the final pachytene substage. However, synaptonemal complexes failed to desynapse and normal diplotene spermatocytes were not observed. Metaphase spermatocytes were not seen in tissue sections from testes of Hsp70-2(-/-) mice, and expression of mRNAs and antigens characteristic of late pachytene spermatocytes (e.g., cyclin A1) and development of spermatids did not occur. Thus, HSP70-2 is required for synaptonemal complex desynapsis, and its absence severely impairs the transition of spermatogenic cells through the late meiotic stages and results in apoptosis beginning with the first wave of germ cell development in juvenile mice. PMID- 9409677 TI - Noggin acts downstream of Wnt and Sonic Hedgehog to antagonize BMP4 in avian somite patterning. AB - In the vertebrate embryo, the lateral compartment of the somite gives rise to muscles of the limb and body wall and is patterned in response to lateral-plate derived BMP4. Activation of the myogenic program distinctive to the medial somite, i.e. relatively immediate development of the epaxial muscle lineage, requires neutralization of this lateral signal. We have analyzed the properties of molecules likely to play a role in opposing lateral somite specification by BMP4. We propose that the BMP4 antagonist Noggin plays an important role in promoting medial somite patterning in vivo. We demonstrate that Noggin expression in the somite is under the control of a neural-tube-derived factor, whose effect can be mimicked experimentally by Wnt1. Wnt1 is appropriately expressed in the neural tube. Furthermore, we show that Sonic Hedgehog is able to activate ectopic expression of Noggin resulting in the blocking of BMP4 specification of the lateral somite. Our results are consistent with a model in which Noggin activation lies downstream of the SHH and Wnt signaling pathways. PMID- 9409678 TI - Differential H4 acetylation of paternal and maternal chromatin precedes DNA replication and differential transcriptional activity in pronuclei of 1-cell mouse embryos. AB - In the mouse embryo, transcriptional activation begins during S/G2 phase of the first cell cycle when paternal and maternal chromatin are still in separate nuclear entities within the same cytoplasm. At this time, the male pronucleus exhibits greater transcriptional activity than the female pronucleus. Since acetylation of histones in the nucleosome octamer exerts a regulatory influence on gene expression, we investigated changes in histone acetylation during the remodeling of paternal and maternal chromatin from sperm entry through to minor genome activation and mitosis. We found (1) neither mature sperm nor metaphase II maternal chromatin stained for hyperacetylated histone H4; (2) immediately following fertilization, hyperacetylated H4 was associated with paternal but not maternal chromatin while, in parthenogenetically activated oocytes, maternal chromatin became hyperacetylated; (3) in zygotes, differential levels and patterns of hyperacetylated H4 between male and female pronuclei persisted throughout most of G1 with histone deacetylases and acetyltransferases already active at this time; (4) when transcriptional differences are observed in S/G2, male and female pronuclei have equivalent levels of H4 hyperacetylation and DNA replication was not required to attain this equivalence and (5) in contrast to the lack of H4 hyperacetylation on gametic chromatin, chromosomes at the first mitosis showed distinct banding patterns of H4 hyperacetylation. These results suggest that sperm chromatin initially out-competes maternal chromatin for the pool of hyperacetylated H4 in the oocyte, that hyperacetylated H4 participates in the process of histone-protamine exchange in the zygote, and that differences in H4 acetylation in male and female pronuclei during G1 are translated across DNA replication to transcriptional differences in S/G2. Prior to fertilization, neither paternal nor maternal chromatin show memory of H4 hyperacetylation patterns but, by the end of the first cell cycle, before major zygotic genome activation at the 2-cell stage, chromosomes already show hyperacetylated H4 banding patterns. PMID- 9409679 TI - Essential roles of the winged helix transcription factor MFH-1 in aortic arch patterning and skeletogenesis. AB - Mesenchyme Fork Head-1 (MFH-1) is a forkhead (also called winged helix) transcription factor defined by a common 100-amino acid DNA-binding domain. MFH-1 is expressed in non-notochordal mesoderm in the prospective trunk region and in cephalic neural-crest and cephalic mesoderm-derived mesenchymal cells in the prechordal region of early embryos. Subsequently, strong expression is localized in developing cartilaginous tissues, kidney and dorsal aortas. To investigate the developmental roles of MFH-1 during embryogenesis, mice lacking the MFH-1 locus were generated by targeted mutagenesis. MFH-1-deficient mice died embryonically and perinatally, and exhibited interrupted aortic arch and skeletal defects in the neurocranium and the vertebral column. Interruption of the aortic arch seen in the mutant mice was the same as in human congenital anomalies. These results suggest that MFH-1 has indispensable roles during the extensive remodeling of the aortic arch in neural-crest-derived cells and in skeletogenesis in cells derived from the neural crest and the mesoderm. PMID- 9409680 TI - Two signalling pathways specify localised expression of the Broad-Complex in Drosophila eggshell patterning and morphogenesis. AB - The Drosophila eggshell, which has a pair of chorionic appendages (dorsal appendages) located asymmetrically along both the anterior/posterior and dorsal/ventral axes, provides a good model to study signal instructed morphogenesis. We show that the Broad-Complex, a gene encoding zinc-finger transcription factors, is essential for the morphogenesis of dorsal appendages and is expressed in a bilaterally symmetrical pattern in the lateral-dorsal anterior follicle cells during late oogenesis. This is induced and specified along the dorsoventral axis by an epidermal growth factor receptor signalling pathway, which includes a localised transforming growth factor-alpha like molecule, Gurken, in the oocyte and the Drosophila EGF receptor homologue, Torpedo, in the surrounding somatic follicle cells. Furthermore, the precisely localised expression of BR-C along the AP axis requires a separate signalling pathway, initiated by a transforming growth factor-beta homologue, Decapentaplegic, in nearby follicle cells. These two signalling pathways, one from the oocyte and the other from the follicle cells, co-ordinately specify patches of follicle cells to express the Broad-Complex in a unique position in respect to both major axes, which in turn directs the differentiation of the dorsal appendages in the correct position on the eggshell. PMID- 9409681 TI - Green Fluorescent Protein in the sea urchin: new experimental approaches to transcriptional regulatory analysis in embryos and larvae. AB - The use of Green Fluorescent Protein (GFP) as a reporter for expression transgenes opens the way to several new experimental strategies for the study of gene regulation in sea urchin development. A GFP coding sequence was associated with three different previously studied cis-regulatory systems, viz those of the SM50 gene, expressed in skeletogenic mesenchyme, the CyIIa gene, expressed in archenteron, skeletogenic and secondary mesenchyme, and the Endo16 gene, expressed in vegetal plate, archenteron and midgut. We demonstrate that the sensitivity with which expression can be detected is equal to or greater than that of whole-mount in situ hybridization applied to detection of CAT mRNA synthesized under the control of the same cis-regulatory systems. However, in addition to the important feature that it can be visualized nondestructively in living embryos, GFP has other advantages. First, it freely diffuses even within fine cytoplasmic cables, and thus reveals connections between cells, which in sea urchin embryos is particularly useful for observations on regulatory systems that operate in the syncytial skeletogenic mesenchyme. Second, GFP expression can be dramatically visualized in postembryonic larval tissues. This brings postembryonic larval developmental processes for the first time within the easy range of gene transfer analyses. Third, GFP permits identification and segregation of embryos in which the clonal incorporation of injected DNA has occurred in any particular desired region of the embryo. Thus, we show explicitly that, as expected, GFP transgenes are incorporated in the same nuclei together with other transgenes with which they are co-injected. PMID- 9409682 TI - maelstrom is required for an early step in the establishment of Drosophila oocyte polarity: posterior localization of grk mRNA. AB - We describe a mutant, maelstrom, that disrupts a previously unobserved step in mRNA localization within the early oocyte, distinct from nurse-cell-to-oocyte RNA transport. Mutations in maelstrom disturb the localization of mRNAs for Gurken (a ligand for the Drosophila Egf receptor), Oskar and Bicoid at the posterior of the developing (stage 3-6) oocyte. maelstrom mutants display phenotypes detected in gurken loss-of-function mutants: posterior follicle cells with anterior cell fates, bicoid mRNA localization at both poles of the stage 8 oocyte and ventralization of the eggshell. These data are consistent with the suggestion that early posterior localization of gurken mRNA is essential for activation of the Egf receptor pathway in posterior follicle cells. Posterior localization of mRNA in stage 3-6 oocytes could therefore be one of the earliest known steps in the establishment of oocyte polarity. The maelstrom gene encodes a novel protein that has a punctate distribution in the cytoplasm of the nurse cells and the oocyte until the protein disappears in stage 7 of oogenesis. PMID- 9409683 TI - Steroid regulated programmed cell death during Drosophila metamorphosis. AB - During insect metamorphosis, pulses of the steroid hormone 20-hydroxyecdysone (ecdysone) direct the destruction of obsolete larval tissues and their replacement by tissues and structures that form the adult fly. We show here that larval midgut and salivary gland histolysis are stage-specific steroid-triggered programmed cell death responses. Dying larval midgut and salivary gland cell nuclei become permeable to the vital dye acridine orange and their DNA undergoes fragmentation, indicative of apoptosis. Furthermore, the histolysis of these tissues can be inhibited by ectopic expression of the baculovirus anti-apoptotic protein p35, implicating a role for caspases in the death response. Coordinate stage-specific induction of the Drosophila death genes reaper (rpr) and head involution defective (hid) immediately precedes the destruction of the larval midgut and salivary gland. In addition, the diap2 anti-cell death gene is repressed in larval salivary glands as rpr and hid are induced, suggesting that the death of this tissue is under both positive and negative regulation. Finally, diap2 is repressed by ecdysone in cultured salivary glands under the same conditions that induce rpr expression and trigger programmed cell death. These studies indicate that ecdysone directs the death of larval tissues via the precise stage- and tissue-specific regulation of key death effector genes. PMID- 9409684 TI - PDP1, a novel Drosophila PAR domain bZIP transcription factor expressed in developing mesoderm, endoderm and ectoderm, is a transcriptional regulator of somatic muscle genes. AB - In vertebrates, transcriptional control of skeletal muscle genes during differentiation is regulated by enhancers that direct the combinatorial binding and/or interaction of MEF2 and the bHLH MyoD family of myogenic factors. We have shown that Drosophila MEF2 plays a role similar to its vertebrate counterpart in the regulation of the Tropomyosin I gene in the development of Drosophila somatic muscles, however, unlike vertebrates, Drosophila MEF2 interacts with a muscle activator region that does not have binding sites for myogenic bHLH-like factors or any other known Drosophila transcription factors. We describe here the isolation and characterization of a component of the muscle activator region that we have named PDP1 (PAR domain protein 1). PDP1 is a novel transcription factor that is highly homologous to the PAR subfamily of mammalian bZIP transcription factors HLF, DBP and VBP/TEF. This is the first member of the PAR subfamily of bZIP transcription factors to be identified in Drosophila. We show that PDP1 is involved in regulating expression of the Tropomyosin I gene in somatic body-wall and pharyngeal muscles by binding to DNA sequences within the muscle activator that are required for activator function. Mutations that eliminate PDP1 binding eliminate muscle activator function and severely reduce expression of a muscle activator plus MEF2 mini-enhancer. These and previous results suggest that PDP1 may function as part of a larger protein/DNA complex that interacts with MEF2 to regulate transcription of Drosophila muscle genes. Furthermore, in addition to being expressed in the mesoderm that gives rise to the somatic muscles, PDP1 is also expressed in the mesodermal fat body, the developing midgut endoderm, the hindgut and Malpighian tubules, and the epidermis and central nervous system, suggesting that PDP1 is also involved in the terminal differentiation of these tissues. PMID- 9409685 TI - A Hedgehog activity gradient contributes to AP axial patterning of the Drosophila wing. AB - The secreted protein Hedgehog (Hh) transmits a signal from posterior to anterior cells that is essential for limb development in insects and vertebrates. In Drosophila, Hh has been thought to act primarily to induce localized expression of Decapentaplegic and Wingless which in turn relay patterning cues at long range. We report here that Hh plays an additional role in patterning the wing. By replacing endogenous Hh activity with that of a membrane-tethered form of Hh, we show that Hh acts directly to pattern the central region of the wing, in addition to its role as an inducer of Dpp. Comparing the biological activities of secreted and membrane-tethered Hh provides evidence that Hh forms a local concentration gradient and functions as a concentration-dependent morphogen in the fly wing. PMID- 9409686 TI - Interactions between the EGF receptor and DPP pathways establish distinct cell fates in the tracheal placodes. AB - The formation of the tracheal network in Drosophila is driven by stereotyped migration of cells from the tracheal pits. No cell divisions take place during tracheal migration and the number of cells in each branch is fixed. This work examines the basis for the determination of tracheal branch fates, prior to the onset of migration. We show that the EGF receptor pathway is activated by localized processing of the ligand SPITZ in the tracheal placodes and is responsible for the capacity to form the dorsal trunk and visceral branch. The DPP pathway, on the contrary, is induced in the tracheal pit by local presentation of DPP from the adjacent dorsal and ventral ectodermal cells. This pathway patterns the dorsal and lateral branches. Elimination of both pathways blocks migration of all tracheal branches. Antagonistic interactions between the two pathways are demonstrated. The opposing activities of two pathways may refine the final determination of tracheal branch fates. PMID- 9409687 TI - Problems in the evaluation of new devices for coronary intervention: what have we learned since 1989? AB - The objectives of this study are to review the problems associated with the evaluation of new devices, the progress made in that evaluation process since 1989, and the role played by the New Approaches to Coronary Intervention (NACI) registry. In 1988-1989, the first wave of new coronary devices (stents, atherectomy, laser catheters) were entering clinical investigation. It seemed unlikely that the small manufacturer-run registries used to gain approval for earlier balloon catheters would be adequate to evaluate the host of complex new devices, each of which might be used for a restricted set of anatomic indications. Moreover, the wide range of arbitrary definitions then in use for fundamental outcomes (such as success, complication, and restenosis), effectively precluded meaningful device-to-device comparisons. Against this backdrop, the NACI registry was formed with National Heart, Lung, and Blood Institute funding to provide an independent and standardized evaluation of the first 8 new devices under evaluation in the United States, across the broad range of their application. The registry employed a unique modular form set to track the sequence of events during complex cases in which serial new devices and balloon angioplasty might be used, either in a planned way, or an unplanned way (to treat complications or suboptimal results). Outcomes were subjected to standardized criteria for (1) the reason for device use (planned, unplanned); (2) success (device, lesion, and procedural success); (3) complications (a) major (death, Q wave myocardial infarction, and emergency coronary artery bypass grafting); or (b) other (groin complications, non-wave myocardial infarction, etc.); and (4) clinical restenosis (any subsequent revascularization, target lesion revascularization). Separate funding for an angiographic core laboratory was obtained in 1992, which analyzed 3,936 (88.9%) of the 4,429 films obtained on patients enrolled between November 1990 and March 1994. The NACI registry has addressed a broad range of problems inherent in the evaluation of new devices for coronary intervention. Whereas the approval process has moved progressively towards randomized clinical trials (and away from registries), the NACI registry offers a unique view of current practice, outside the narrow scope of the limited number of randomized trials that have been performed to date. This article shows, however, that we have learned about more than the devices themselves since 1989 we have also learned about the importance of knowing the reason for device use, using precise definitions of endpoint variables, understanding the financial and reimbursement ramifications of new device trials, and upholding strict investigator ethics during the conduct of such evaluations. PMID- 9409688 TI - New Approaches to Coronary Intervention (NACI) registry: history and methods. AB - The New Approaches to Coronary Intervention (NACI) registry was funded in 1990 by the National Heart, Lung, and Blood Institute to evaluate the safety and efficacy of new coronary interventional devices. The registry collected data from 39 clinical centers on 8 new devices: 3 atherectomy devices, 2 stents, 2 laser devices and the laser balloon. The original funding called for a total of 500 patients to be recruited for each device. One device (the laser balloon) was removed from use early in the recruitment period. Subsequent funding was obtained for an additional 500 patients treated with directional atherectomy, and 500 additional female patients who were recruited when the main recruitment had finished. When all recruitment was finished in March 1994, a total of 4,429 patients had been recruited. Consecutive patients treated with a new device at each clinical site were recruited regardless of underlying clinical condition or result. Patients were followed for 1 year by telephone contact at 6 weeks, 6 months and 1 year. Overall 1-year follow-up compliance was excellent at 96% of patients. The NACI database was designed to allow analysis of the complex relationships between patient, lesion and device. In particular, the mode of use of each device, the treatment order, and the outcome after each device attempt were recorded. All in-hospital clinical events were recorded as well as an assessment of the success of the procedure by the operator. The results of each telephone follow-up included any interim cardiac events, anginal status, and results of any subsequent angiograms or exercise test. Any repeat percutaneous procedures or bypass surgery on an NACI patient were recorded in the same detail as the original procedure. All data collected were subjected to extensive editing at the coordinating center to ensure both internal and external consistency. An angiographic core laboratory was also funded for NACI in 1992 to provide uniform analyses of procedural angiograms. Measurements were made at each step of the procedure, that is preprocedure, after each device use, and postprocedure. Both quantitative and qualitative measurements were made at each step using standardized definitions, enabling a complete description of the procedure to be analyzed. Angiograms were made available to the core laboratory for 89% of all initial procedures. PMID- 9409689 TI - Reliability of the quantitative angiographic measurements in the New Approaches to Coronary Intervention (NACI) registry: a comparison of clinical site and repeated angiographic core laboratory readings. AB - To assess the agreement of clinical site and angiographic core laboratory readings obtained in the New Approaches to Coronary Intervention (NACI) registry, we reviewed the angiographic results obtained in 787 lesions assessed both by the sites and the core laboratory, including 135 lesions analyzed twice (> or =2 months apart) by the angiographic core laboratory. Although moderate agreement was demonstrated between the clinical site and angiographic core laboratory for qualitative lesion morphology such as lesion calcium (kappa [kappa] = 0.42), only fair agreement was found between site and core laboratory estimation of lesion ulceration (kappa = 0.33); thrombus (kappa = 0.30); and eccentricity (kappa = 0.27); with poor agreement for angulation (kappa = 0.16); and proximal vessel tortuosity (kappa = 0.03). Agreement for qualitative morphology was better for repeated core laboratory readings of lesion eccentricity (kappa = 0.75); angulation (kappa = 0.72); thrombus (kappa = 0.68); proximal vessel tortuosity (kappa = 0.66); and calcification (kappa = 0.64). Quantitative angiographic measurements correlated moderately between the clinical site using the digital caliper method and the core laboratory using the automated edge-detection method, including preprocedural percentage diameter stenosis (intraclass correlation [R] = 0.50) and postprocedural percentage diameter stenosis (R = 0.63). Repeated core laboratory readings had almost perfect agreement, with R ranging from 0.88 for postprocedural percentage diameter stenosis to 0.93 for reference vessel diameter and pre- and postprocedural minimal lumen diameters. Repeated angiographic core laboratory readings provided highly consistent quantitative and qualitative morphologic results in the NACI registry, but the core laboratory readings varied substantially from those obtained at the clinical site. More standardized angiographic analytic criteria and core laboratory feedback to investigators may improve agreement between the clinical sites and the angiographic core laboratory in subsequent studies. PMID- 9409690 TI - Influence of gender on in-hospital clinical and angiographic outcomes and on one year follow-up in the New Approaches to Coronary Intervention (NACI) registry. AB - Higher complication rates and lower success rates for treatment of women compared with men have been reported in prior studies of coronary angioplasty and in most early reports of outcome with new coronary interventional devices. In multivariate analysis this has been attributed largely to older age and other unfavorable clinical characteristics. These results are reflected in the current guidelines for coronary angioplasty. Women in prior studies have also had different distributions of vessel and lesion characteristics, but the influence of these differences on the outcome of new-device interventions have not been adequately evaluated. This article evaluates the influence of gender on clinical and angiographic characteristics, interventional procedure and complications, angiographic success, and clinical outcomes at hospital discharge and 1-year follow-up, as observed in the New Approaches to Coronary Intervention (NACI) registry. The NACI registry methodology has been reported in detail elsewhere in this supplement. This study focuses on the 90% of patients-975 women and 1,880 men-who had planned procedures with a single new device and also had angiographic core laboratory readings. Women compared with men were older, had more recent onset of coronary ischemic pain that was more severe and unstable, and had more frequent histories of other adverse clinical conditions. The distributions of several but not all angiographic characteristics before intervention were considered more favorable to angioplasty outcome in women. Differences were observed in device use and procedure staging. Angiographically determined average gain in lumen diameter after new-device intervention, with or without balloon angioplasty, was significantly less in women (1.38 mm) than in men (1.53 mm; p < 0.001); this 0.15 mm difference is consistent with the 0.16-mm smaller reference vessel lumen diameter of women. However, final percent diameter stenoses and TIMI flow and lesion compliance characteristics were similar. Among procedural complications, only treatment for hypotension, blood transfusion, and vascular repair occurred more often in women. More women than men were clinically unstable (2.1% vs 1.1%) or went directly to emergent coronary artery bypass graft surgery (CABG; 1.2% vs 0.6%) on leaving the interventional laboratory. However, in hospital death (1.4% vs 1.1%), Q-wave myocardial infarction (MI) (0.9% vs 1.1%), and emergent CABG (1.5% vs 1.0%, for women and men, respectively) were not significantly different. Nonemergent CABG was more frequent in women (1.8% vs 0.9%; p < 0.05) and length of hospital stay after device intervention was longer (4.4 days vs 3.8 days in men; p < 0.01). In both univariate and multivariate analyses gender did not emerge as a significant variable in relation to the combined endpoint, death, Q-wave MI, or emergent CABG at hospital discharge. At 1 year follow-up more women than men reported improvement in angina (70% vs 62%) and fewer women than men had had repeat revascularization (32% vs 36%). Similar proportions were alive and free of angina, Q-wave MI and repeat revascularization (46% of women vs 45% of men). Although several procedure-related complications were more frequent in women than men after coronary interventions with new devices, no important disadvantages were observed for women in the rates of major clinical events at hospital discharge and at 1-year clinical follow-up. Additional studies are needed to evaluate the complex interplay of clinical, vessel, and lesion characteristics on success and complications of specific interventional techniques and to determine whether gender, per se, is a risk factor and whether gender specific interventional strategies may be beneficial. PMID- 9409691 TI - Frequency and predictors of major in-hospital ischemic complications after planned and unplanned new-device angioplasty from the New Approaches to Coronary Intervention (NACI) registry. AB - The purpose of this study was to determine the frequency and predictors of major in-hospital ischemic complications after planned and unplanned procedures with new angioplasty devices from the New Approaches to Coronary Intervention (NACI) registry. The NACI registry is a multicenter, voluntary reporting of consecutive patients undergoing new-device angioplasty procedures using atherectomy catheters, stents, or lasers. This registry affords the opportunity to evaluate the performance of new angioplasty devices during elective and urgent circumstances. The study population consisted of 3,340 patients with 3,733 lesions (2,921 in native coronary arteries and 812 in saphenous vein grafts [SVGs], who were treated with new devices over a 3.5-year period and had their angiograms analyzed independently at a central angiographic core laboratory. Their in-hospital course and multivariate predictors of the complications in planned and unplanned procedures, further divided into native and SVG lesions, were evaluated. In 82.2% of native coronary artery lesions and 96.9% of SVG lesions, the procedure with a device had been planned due to unfavorable lesion characteristics for PTCA. In the remaining lesions, device use was unplanned, and was performed mainly to treat a suboptimal result (59-80.4%) after percutaneous transluminal coronary angioplasty (PTCA), and less frequently after important complications of PTCA including abrupt closure and PTCA failure. In native artery cohort, major in-hospital ischemic complications (death, Q-wave myocardial infarction [MI], or emergency coronary artery bypass surgery) occurred in 2.7% of the planned and 9.9% of the unplanned procedures (p < 0.001), whereas in SVG such complications occurred in 3.6% of the planned and 8.7% of unplanned procedures (p = 0.21). Multivariate analysis revealed several predictors of major ischemic complications from planned native coronary artery device use: post-MI angina (odds ratio = 2.83); severe concomitant noncardiac disease (odds ratio = 2.5); multivessel disease (odds ratio = 1.75); and de novo lesions (odds ratio = 2.3). Multivariate predictors of major complications in unplanned native coronary artery procedures included high surgical risk (odds ratio = 3.08), and tortuous lesion (odds ratio = 2.41). In SVG lesions, the independent predictors of major complications for planned procedures included age (odds ratio = 1.09), high surgical risk (odds ratio = 4.34), and thrombus (odds ratio = 2.62). In native and SVG lesions, rates of major complications of planned procedures was acceptable (2.7-3.67%), but unplanned use of a new device was associated with a significantly higher rate of in-hospital complications (approximately 9%). Multivariate predictors for major ischemic complications included both clinical and lesion characteristics, and differed for native versus SVG lesions, as well as for planned versus unplanned procedures. PMID- 9409692 TI - Directional coronary atherectomy (DCA): a report from the New Approaches to Coronary Intervention (NACI) registry. AB - Directional coronary atherectomy (DCA) with the Simpson coronary atherocath seeks to debulk rather than simply displace obstructive tissue and is a means of enlarging the stenotic coronary lumen. This report from the New Approaches to Coronary Intervention (NACI) registry describes the experience of 1,196 patients who underwent DCA as the sole treatment for either native vessel or vein graft lesions. Device success (post-DCA residual stenosis <50% and > or =20% improvement) was achieved in 87.8%, with a lesion success rate (postprocedural residual stenosis <50% and > or =20% improvement) of 94.0%. The mean resultant stenosis after all interventions (by core laboratory) was 19%. Significant in hospital complications occurred in 2.8% of patients with DCA attempts, including death 0.6%, Q-wave myocardial infarction (MI) 1.5%, and emergent coronary artery bypass graft surgery (CABG) 2.8%. At 1-year follow-up, cumulative mortality was 3.6%, with repeat revascularization in 28% (repeat percutaneous transluminal coronary angioplasty, 20.1%; CABG, 10.6%). This reflected percutaneous or surgical revascularization of the original lesion (target lesion revascularization) in 22.6% of patients. Subgroup analysis showed a lower lesion success rate and an increased complication rate for unplanned use, vein graft treatment, and treatment of a de novo (vs a restenotic) lesion. Multivariate analysis shows that diabetes mellitus, unstable angina, treatment of a restenotic lesion, and greater residual stenosis after the initial procedure were independent predictors of the composite endpoint of death/Q-wave MI/target lesion revascularization by 1-year follow-up. Among these generally favorable acute and 1-year results, the NACI directional atherectomy data confirm the "bigger is better" hypothesis: that lesions with a lower residual stenosis after a successful procedure had significantly fewer target lesion revascularizations between 30 days and 1 year, with no increase in major adverse events. PMID- 9409693 TI - High-speed rotational atherectomy of human coronary stenoses: acute and one-year outcomes from the New Approaches to Coronary Intervention (NACI) registry. AB - High-speed rotational atherectomy (RA) is a new percutaneous procedure for treatment of coronary stenoses that operates by the unique mechanism of plaque abrasion. This article reports acute (in-hospital) outcomes and 1-year follow-up in a large cohort of patients treated with this device by NACI investigators. A total of 525 patients with 670 lesions treated with RA form the substrate of this report. Patients tended to be older (mean age 64.8 years) than those in previously reported series of percutaneous transluminal coronary angioplasty (PTCA), with more extensive disease and more complex lesions. Calcification was present in 54% of lesions, and eccentricity in 41%. Balloon angioplasty postdilation was performed after RA in 88% of cases. Angiographic and procedural success (angiographic success without death, Q-wave myocardial infarction [MI] or emergency coronary artery bypass graft [CABG] surgery) rates were 89% and 88%, respectively. Acute in-hospital events included 4 deaths (1%) and 1 emergency CABG surgery (0.4%). MI occurred in 6% of patients, consisting predominantly of non-Q-wave MI (5%). After RA, angiographic complications included coronary dissection (12%), abrupt closure (5%), side branch occlusion (3%), and distal embolization (3%). Most of these were resolved after postdilation except for coronary dissection, which was present in 15% of lesions treated. Mean length of stay was 3 days. At 1-year follow-up, 27% of patients required target lesion revascularization and 30% had experienced death, Q-wave MI, or target lesion revascularization. Preprocedural characteristics that independently predicted 1 year death, Q-wave MI, or target lesion revascularization were male gender, high risk for surgery, target lesions that were proximal to or in bifurcations, eccentric, long, or highly stenosed. RA, even when applied to lesions of traditionally unfavorable morphology, appears to provide reasonable procedural and angiographic success rates. Restenosis and progression of disease contribute to subsequent clinical and procedural events. PMID- 9409694 TI - Predictors of acute and long-term outcome with transluminal extraction atherectomy: the New Approaches to Coronary Intervention (NACI) registry. AB - The New Approaches to Coronary Intervention (NACI) registry was established to define the role of new coronary devices in overcoming the limitations of balloon angioplasty. The purpose of the present study was to evaluate the acute and long term efficacy of the transluminal extraction catheter (TEC) device utilizing data from the NACI registry and identify clinical and anatomic patient subsets who may benefit from this device. From 1990-1994, >4,300 patients from 39 clinical sites enrolled consecutive patients treated with one of the 7 new devices to the NACI registry. The study population consists of 331 patients (385 lesions) treated with planned TEC as the sole new device. Of these patients, 243 (292 lesions) were treated for saphenous vein graft (SVG) disease and 88 (93 lesions) for native disease. Patients undergoing SVG treatment were older and more likely to be male. They had lower ventricular function, more unstable angina, and a higher incidence of congestive heart failure. Multivessel disease was more prevalent in the SVG cohort, as was evidence of thrombus before treatment. Although device success was achieved in 50% of SVG lesions and 41% of native lesions, lesion success was achieved in 90% and 78%, respectively, after adjunctive balloon angioplasty, and procedure success rates were 86% and 79%, respectively. The in hospital major complication (death/Q-wave myocardial infarction/emergency coronary artery bypass graft [CABG] surgery) rate was higher in the SVG cohort (6.2% vs 2.3%), mainly due to higher mortality rate (5.3% vs 1.1%). Multivariate analysis showed that SVG was not an independent predictor for either an in hospital major complication or clinical failure. The risk factors for major in hospital complications were history of congestive heart failure (odds ratio = 3.17) and thrombus (odds ratio = 3.36). For clinical failure the risk factors were diabetes (odds ratio = 1.88), thrombus (odds ratio = 2.08), and calcium (odds ratio = 3.09). One-year rates of death, Q-wave myocardial infarction, or any repeat revascularization were 51% in the SVG cohort and 41% in the native cohort. Following adjustment, patients treated for SVG disease did not have a higher risk when compared with those treated for native disease. The factors significantly associated with this composite event at 1 year are male (relative risk = 1.41), patients with history of congestive heart failure (relative risk = 1.56), and total occlusions (relative risk = 1.52). This study shows that for both SVG and native cohorts, device success rates were low with TEC alone, but acceptable lesion success rates were achieved when adjunctive PTCA was used. In hospital as well as 1-year major complications were higher in the SVG cohort. However, after adjusting for other risk factors, SVG attempt was not significantly associated with either in-hospital or 1-year events. PMID- 9409695 TI - Acute and long-term outcome after Palmaz-Schatz stenting: analysis from the New Approaches to Coronary Intervention (NACI) registry. AB - The randomized Stent Restenosis Study (STRESS) and Belgium Netherlands Stent (Benestent) trials established that elective use of Palmaz-Schatz stents (PSSs) in native coronary arteries with de novo lesions is associated with increased procedural success and reduced restenosis. However there are other clinical indications for which stents are commonly used (unplanned use, vein grafts, restenosis lesions) that are not addressed in these studies. From 1990-1992, 688 lesions in 628 patients were treated with PSSs in the New Approaches to Coronary Intervention (NACI) registry. Angiographic core laboratory readings were available for 543 patients (595 lesions, of which 106 were stented for unplanned indications, 239 were in saphenous vein bypass grafts, and 296 were previously treated). The cohort of patients in whom stents were placed for unplanned indications had more women, current smokers, and had a higher incidence of recent myocardial infarction (MI). Patients who underwent stenting of saphenous vein grafts were older, had a higher incidence of diabetes mellitus, unstable angina, prior MI, and congestive heart failure. Lesion success was similar in all cohorts (98%), but procedural success was significantly higher for planned stenting (96% vs 87%; p < 0.01). Predictors of adverse events in-hospital were presence of a significant left main stenosis and stenting for unplanned indication. The incidence of target lesion revascularization by 30 days was significantly higher for patients undergoing unplanned stenting due to a higher risk for stent thrombosis. Recent MI, stenting in native lesion, and small postprocedural minimum lumen diameter independently predicted target lesion revascularization at 30 days. Independent predictors of death, Q-wave myocardial infarction, or target lesion revascularization at 1 year included severe concomitant disease, high risk for surgery, left main disease, stenting in the left main coronary artery, and low postprocedure minimum lumen diameter. PMID- 9409696 TI - The use of the Gianturco-Roubin intracoronary stent: the New Approaches to Coronary Intervention (NACI) registry experience. AB - The objective of this study is to compare the in-hospital and follow-up outcome in patients receiving the Gianturco-Roubin stent (GRS) who were enrolled in the New Approaches to Coronary Intervention (NACI) registry. The GRS was approved by the US Food and Drug Administration (FDA) in August 1992 for the treatment of acute or threatened closure after a percutaneous intervention. The application of intracoronary stenting has broadened substantially in the last few years, but less is known about the use of this device for other indications. Since the NACI registry includes patients stented for other indications, a comparison of these groups with patients being stented for acute or threatened closure was undertaken. A GRS was deployed in 497 NACI registry patients. Of these, 466 patients received a GRS in 1 of 3 of the following ways: (1) 351 unplanned stenting after conventional angioplasty of the same lesion; (2) 54 after failed/suboptimal use of a new device in the same lesion; and (3) 61 in planned stenting procedures. This analysis focuses on these 3 patient subgroups and compares their in-hospital outcome and subsequent follow-up to 1 year. There were 520 stented segments in the 466 patients. The group with stenting after failed/suboptimal new-device use had a higher incidence of myocardial infarction (MI) and cardiogenic shock than either the patients with unplanned stenting after percutaneous transluminal coronary angioplasty (PTCA) or planned stenting (MI 22.2% vs 12.0% vs 0%, respectively, and cardiogenic shock 5.6% vs 0.9% vs 0%, respectively; p < 0.05). This group also had significantly lower procedural success (58.7% vs 75.3% vs 81.5%, respectively; p < 0.05). Although not statistically significant, the requirement for transfusion was higher in the unplanned and new-device stented groups than in the planned group (10.5% vs 16.7% vs 1.6%, respectively). Likewise, the incidence of Q-wave MI was higher in the new-device group (22.2% vs 12% vs 0%, respectively; p < 0.05). Despite a higher, in-hospital complication rate in the unplanned groups, follow-up from discharge to 1 year showed similar outcome. In particular, percutaneous reintervention of the stented segment occurred in: 13.0% in the unplanned after new device; 17.4% in the unplanned after PTCA; and 26.2% in the planned group. Although not statistically significant, the higher incidence of percutaneous target lesion revascularization in the planned group probably represents the greater incidence of restenotic lesions in this cohort. In this very heterogeneous group of patients, including those with failure of another new device, the use of the GRS is associated with acceptable in-hospital and follow-up complication rates, although complications were clearly greater when unplanned use of the stent was needed, particularly after failure of another new device. Although the experience is small, patients having the GRS placed in an elective fashion, i.e., the planned group, appear to experience lower in-hospital complication rates, although they have a higher rate of subsequent target lesion revascularization, in this group of predominantly restenotic lesions. PMID- 9409697 TI - Excimer laser coronary angioplasty: the New Approaches to Coronary Intervention (NACI) experience. AB - In the New Approaches to Coronary Intervention (NACI) registry, 887 patients were electively treated with excimer laser coronary angioplasty (ELCA) for coronary artery disease. The Advanced Interventional System (AIS) system was used in 487 cases; the Spectranetics system, in 400. The mean age was 63.4 years. Most patients had unstable angina (60.3%); 43.7% had a prior myocardial infarction; and 18.6% were high risk or inoperable patients. Mean ejection fraction was 55.4%. A total of 1,000 lesions were treated in the 887 patients. Of the 1,000 lesions treated with ELCA in the 887 patients, 36% were in the right coronary artery; 33%, left anterior descending; 13%, circumflex; 3%, left main; and 16.6%, vein graft. By angiographic core laboratory analysis available for 752 (85%) patients with 839 lesions, lesions were 12.76 mm long. The minimum lumen diameter increased to 1.29 mm after the laser and finally to 1.95 mm after adjunctive percutaneous transluminal coronary angioplasty (PTCA) (which was performed in 93% of all lesions), with a final residual stenosis of 32.1% and Thrombolysis in Myocardial Infarction (TIMI) grade 3 flow in 95%. Dissections of grades B, C, or D were seen after 22.0% of initial laser attempts, and postlaser perforations were noted in 2.6%. Additional such dissections accumulated after adjunctive PTCA but the perforation rate remained low. Procedural success was achieved in 84% of patients, but 1.2% died, 0.7% experienced Q-wave myocardial infarction (MI), and 2.7% required emergency bypass surgery. Multiple logistic regression analysis could not identify any independent predictors of these in-hospital complications. One-year mortality was 5.7% and the cumulative incidence of Q-wave MI was 1.5%. Coronary artery bypass graft (CABG) surgery was performed in 15.0% of patients whereas 25.5% required repeat percutaneous intervention with a target lesion revascularization rate of 31%. Independent predictors of death, Q-wave MI, or target lesion revascularization (which, combined, occurred in 35.6% of patients) were the absence of prior MI, ELCA in the circumflex, perforation after the procedure, and small (<2 mm) final minimal lumen diameter. Considering the large number of patients with high-risk lesions, laser angioplasty was performed with excellent procedural success rates and a reasonable incidence of major complications. PMID- 9409698 TI - Identification of the midbrain locomotor nuclei and their descending pathways in the teleost carp, Cyprinus carpio. AB - In order to identify the mesencephalic spinal pathways for initiation of swimming in the carp, we employed electrical and chemical microstimulation of the mesencephalic tegmentum. Electrical stimulation of the midbrain in decerebrate carp produced bilateral or unilateral rhythmic movements of the tail. Bilateral alternating movements were induced by stimulation with the lowest threshold currents to the brain region just beneath the third ventricle at the level of the mid mesencephalon. The region included the nucleus of medial longitudinal fasciculus (Nflm), the medial longitudinal fasciculus (flm), the red nucleus (Nrb). To specify the nuclei of the origin of the descending pathway, we microinjected 0.1 M L-glutamic acid to the region. Both bilateral and unilateral tail movements were induced, the majority being the latter. The unilateral movements were accompanied with tail flips toward the ipsilateral side of stimulation sites. The smallest injection volume required for initiation of the movement was recorded at the Nflm. Bilateral tail movements were produced only by injections into the medial region between the nucleus of the both sides. The present results imply a crucial role of Nflm neurons in the initiation of swimming Nflm neurons on one side project through flm to the ipsilateral spinal cord along its entire length and regulate activities of the individual central pattern generators. PMID- 9409699 TI - Influence of a GABA(B) receptor antagonist on the sleep-waking cycle in the rat. AB - The influence of CGP 35348 (a GABA(B) receptor antagonist) on the sleep-waking cycle was studied in rats. The animals were injected i.p. at the beginning of the light period and the data expressed by 2-h periods and total duration (6 h). At 100 mg/kg, slow-wave sleep (SWS) was decreased during the 6-h recording with a peculiar decrease during the first 2 h. SWS was subdivided into three stages: slow-waves; spindles occurring as SWS deepens; and intermediate stage appearing prior to paradoxical sleep (PS). Only the slow-wave stage and intermediate stage were decreased. Waking was increased during the 6-h recording. It was subdivided into waking with hippocampal theta rhythm (psychomotor active waking) and waking without theta activity (quiet waking). Both were increased during the first 2 h. However, quiet waking was increased throughout the recording duration. At 300 mg/kg, SWS was decreased during the three 2-h periods. This decrease was principally related to a decrease of the slow-wave stage. PS was increased over the 6-h recording with a marked increase during the second 2-h period. Consequently, under the influence of the GABA(B) receptor antagonist, the SWS was decreased at the expense of behavioral stages with cortical low-voltage activity (waking and PS). GABAergic neurons are present in the mesopontine structures responsible for these two stages. We can conclude that endogenous GABA acting at the GABA(B) receptor level participates in the regulation of waking and PS. PMID- 9409700 TI - Selective metabotropic receptor agonists distinguish non-ionotropic glutamate binding sites. AB - Metabotropic glutamate receptors (mGluRs) are thought to mediate diverse processes in brain including synaptic plasticity and excitotoxicity. These receptors are often divided into three groups by their pharmacological profiles. [3H]Glutamate binding in the presence of compounds selective for ionotropic glutamate receptors can be used as a general assay for these receptors; subtypes of this non-ionotropic [3H]glutamate binding differ in both pharmacology and anatomical distribution, and are differentially sensitive to quisqualate. The characteristics of these binding sites are consistent with those of group 1 (high affinity quisqualate) and group 2 (low-affinity quisqualate) mGluRs. Under our assay conditions, no [3H]glutamate binding to group 3-like (L-AP4 sensitive) sites could be demonstrated. We have attempted to characterize particular agents which may selectively measure [3H]glutamate binding to mGluR subtypes. We used two isomers of 2-(carboxycyclopropyl)glycine, L-CCG-I and L-CCG-II, and the (2S,1'R,2'R,3'R) isomer of 2-(2,3-dicarboxycyclopropyl)glycine (DCG-IV) as competitors of non-ionotropic [3H]glutamate binding sites. DCG-IV clearly distinguishes two binding sites. Quantitative levels of DCG-IV binding by anatomic region correlate with quisqualate-defined binding subtypes: high affinity DCG-IV binding correlates with low-affinity quisqualate binding, whereas low-affinity DCG-IV binding correlates with high-affinity quisqualate binding. L CCG-II displaces only one type of non-ionotropic [3H]glutamate binding, corresponding to high-affinity quisqualate binding. Therefore DCG-IV and L-CCG-II at appropriate concentrations appear to distinguish binding to putative group 2 vs. group 1 mGluRs. L-CCG-I displaces both high- and low-affinity quisqualate binding sites, but unlike the other two compounds, does not clearly distinguish between them. PMID- 9409702 TI - Paired helical filaments in corticobasal degeneration: the fine fibrillary structure with NanoVan. AB - Paired helical filaments (PHF) composed of hyperphosphorylated tau proteins are characteristic findings in neurodegenerative disorders, including Alzheimer's disease (AD) and corticobasal degeneration (CBD). The filaments in CBD differ from those in AD by a reduced number of tau isoforms and less stable ultrastructure. To further compare the ultrastructure of both filaments, we employed a novel staining reagent, NanoVan, as well as aurothioglucose and uranyl acetate. With commonly used uranyl acetate, both kinds of filaments appeared as twisted ribbons 15-20-nm and 21-23-nm wide, respectively, without significant internal substructure. With application of aurothioglucose, only few structural details were apparent. With NanoVan, AD filaments showed similar structure to that with uranyl acetate but CBD filaments displayed a highly heterogeneous appearance consistent with the dissociation of the 20-25-nm-wide filaments along two longitudinal axes. This was evident by the presence of thinner, 12-13-nm-wide filaments and filaments that splayed into two 20-25-nm-wide components at one or both ends. Moreover, detection of a prominent, 7-8-nm-wide axial region distinguished up to four protofilaments per one filament. Each protofilament appeared to contain two 3-5-nm-wide fibrils separated by an approximately 1-nm wide axial region. The results suggest that 3-5-nm fibrils are the smallest structural subunits of filaments in CBD and that NanoVan may be an unique reagent in detecting eight-fibril organization in these less stable filaments. PMID- 9409703 TI - Reduced prepulse inhibition after electrolytic lesions of nucleus accumbens subregions in the rat. AB - A variety of neurochemical and anatomical studies have shown that the ventral striatal nucleus accumbens (NAcc) can be divided into a predominantly medial 'shell' and a predominantly lateral 'core'. These accumbens subdivisions are innervated by different cortical regions and project to different anatomical targets. Additionally, recent behavioral observations suggest functional differences between the accumbens shell and core. In order to further examine core and shell function, we measured prepulse inhibition (PPI) of the acoustic startle reflex after electrolytic lesions aimed at the central NAcc, the NAcc shell, the NAcc core, and the adjacent anteroventrolateral striatum. Acoustic startle is the contraction of whole-body musculature in response to a sudden, loud auditory stimulus, and PPI is the inhibition of acoustic startle by a weak auditory 'prepulse' administered 30-500 ms prior to the startling stimulus. Previous work has shown that PPI is regulated by the NAcc, and recent observations suggest that PPI is regulated by different neurochemical interactions in the core and shell. PPI was significantly reduced by electrolytic lesions of the central NAcc, as well as by lesions that predominantly damaged either the NAcc shell or the core. Lesions of the anteroventrolateral striatum and lateral edge of the NAcc core did not significantly alter PPI. These findings suggest that, despite neurochemical differences between the two subregions, PPI is regulated by both the NAcc shell and core. PMID- 9409701 TI - Enhanced plasma DHEAS, brain acetylcholine and memory mediated by steroid sulfatase inhibition. AB - Steroid sulfatase inhibitors can alter the metabolism of neurosteroids which modulate brain function. Administration of the non-steroidal steroid sulfatase inhibitor (p-O-sulfamoyl)-N-tetradecanoyl tyramine (DU-14) to rats for 15 days increased plasma dehydroepiandrosterone sulfate (DHEAS) concentrations by 88.2%, decreased plasma dehydroepiandrosterone (DHEA) concentrations by 84.6%, increased hippocampal acetylcholine (ACh) release determined via in vivo microdialysis by almost 3-fold, and produced a significant blockade of scopolamine-induced amnesia as measured by a passive avoidance test. These results suggest DHEAS rather than DHEA enhances brain cholinergic function and that steroid sulfatase inhibition may become an important tool for enhancing neuronal functions, such as memory, mediated by excitatory neurosteroids. PMID- 9409704 TI - Diazepam-sensitive GABA(A) receptors in the NTS participate in cardiovascular control. AB - The present study employed neuropharmacological and receptor binding protocols to determine if diazepam-sensitive (DS) gamma-aminobutyric acid-A (GABA(A)) receptors in the nucleus tractus solitarius (NTS) participate in autonomic regulation of cardiovascular function. The first set of protocols was designed to determine if GABA(A) receptors in the NTS were functionally modulated by the benzodiazepine agonist, diazepam. Mean arterial pressure and heart rate responses to microinjection of GABAergic substances into the NTS were examined in urethane anesthetized rats. Microinjection of the GABA(A) agonist isoguvacine into the NTS increased mean arterial pressure and heart rate, and these effects were blocked by the GABA(A) receptor antagonist, bicuculline. Preadministration of diazepam into the NTS potentiated the pressor actions of isoguvacine and had variable effects on heart rate changes. Flumazenil, a benzodiazepine antagonist, blocked the diazepam-induced potentiation of the pressor response to isoguvacine. The second protocol employed receptor autoradiography to examine the presence of DS and diazepam-insensitive (DI) GABA(A) receptors in the NTS. Autoradiography confirmed that DS GABA(A) receptors were present in the NTS; however, no measurable levels of DI GABA(A) receptors were detected. We conclude that GABA(A) mediated integration of central autonomic control in the NTS is mediated solely by DS GABA(A) receptors. PMID- 9409705 TI - Neuroprotective effect of a novel AMPA receptor antagonist, YM90K, in rat focal cerebral ischaemia. AB - It has been reported that delayed treatment with alpha-amino-3-hydroxy-5-methyl-4 isoxazole (AMPA) receptor antagonists was able to more completely inhibit glutamate neurotoxicity than N-methyl-D-aspartate (NMDA) receptor antagonists. Therefore, we investigated the neuroprotective effect of YM90K, an AMPA receptor antagonist, on focal cerebral lesions induced by thrombotic middle cerebral artery (MCA) occlusion in rats, particularly in the early phase of the cerebral ischaemic lesions. The MCA was occluded by photochemical reaction between transmural green light and systemically administered Rose Bengal, which causes endothelial injury followed by platelet adhesion, aggregation and formation of a platelet and fibrin-rich thrombus at the site of photochemical reaction. The infarct size was measured at 24 and 72 h after the MCA occlusion by a histochemical technique. YM90K was administered at various doses as a continuous infusion for 4 h, beginning 0 to 3 h after the MCA occlusion. YM90K (10 and 20 mg/kg per h for 4 h continuous infusion), starting immediately after the MCA occlusion significantly (P < 0.05) reduced the infarct size at 24 h after MCA occlusion in a dose-dependent manner. Further, the agent showed the same efficacy at 72 h after. The inhibitory effect of YM90K (20 mg/kg per h) on the infarct size was the same when the drug was started immediately, 1, 2 and 3 h after MCA occlusion. In conclusion, the novel AMPA receptor antagonist YM90K was effective in the treatment of focal cerebral ischaemic lesions. Activation of AMPA receptor may play a key role in the development of cerebral infarct in the early phase of ischaemia in rats. PMID- 9409706 TI - Nitric oxide (NO): in vivo electrochemical monitoring in the dorsal horn of the spinal cord of the rat. AB - NO synthase (NOS) is largely distributed in the superficial and deep laminae of the dorsal horn as well as in dorsal root ganglion cells. It has been proposed that nitric oxide (NO) participates in the transmission of sustained, and possibly brief, nociceptive, inputs at the spinal level. The aim of this study was to check the ability of in vivo electrochemical monitoring of NO within the dorsal horn of the lumbar spinal cord (L3-L4 level) of chloral hydrate anesthetized or decerebrated spinalized rats. 30 microm diameter and 450 microm length treated carbon fiber electrodes coated with nickel(II) tetrakis (3-methoxy 4-hydroxy-phenyl) porphyrine and NafionR, and associated with differential normal pulse voltammetry, gave a peak of oxidation current around 650 mV (vs. Ag-AgCl) in vitro in NO solutions between 0.125 and 1.25 microM. In vivo, a 650 mV peak appeared which was stable (recording interval 2 min) for up to 3 h (+/-6%). Comparison between in vitro calibration and in vivo voltammograms gave an estimated in vivo extracellular concentration of 0.50 microM. In vivo, peaks decreased by 95% at 90 min and for up to 3 h after an i.p. injection of 100 mg/kg of the NOS inhibitor (NOSI) L-arginine-p-nitroanilide (L-ANA). At the same dose i.p., N(G)-nitro-L-arginine methyl ester (L-NAME) was almost ineffective after 90 min in animals paralyzed with pancuronium bromate or gallamine trethiodide. However, in non-curarized decerebrated spinalized animals, L-NAME depressed the voltammograms by 36% at 90 min. S-Ethylthiourea (80 mg/kg i.p.), also decreased the voltammograms by 45% at 140 min, and finally, 7-nitroindazole (7-NI, 90 mg/kg i.p), induced a important decrease of the 650 mV peak (23% of control) at 120 min. These results are in agreement with biochemical data showing the decrease of NOS activity within the lumbar spinal cord by L-NAME (45% of control at 90 min) and 7-NI (20% of control at 90 min). The NO donor hydroxylamine (30 mg/kg i.p.) significantly increased the peaks (140% at 90 min), and sodium nitroprusside (SNP, 20 mM) when directly superfused upon the spinal cord (200-300 microl min( 1)) induced a large increase in the peak (300% at 90 min). Moreover, SNP 60 min after L-ANA, or 90 min after L-NAME, rapidly restored the 650 mV peak up to control values. These results demonstrate the validity of electrochemical monitoring of NO within the dorsal horn of the spinal cord. The in vivo electrochemical detection of NO is in progress to study the implication of this messenger in the transmission of nociceptive messages at the spinal level. PMID- 9409707 TI - Involvement of iron in MPP+ toxicity in substantia nigra: protection by desferrioxamine. AB - Desferrioxamine (DES) protective effect against 1-methyl-4-phenylpyridinium (MPP+) toxicity was evaluated by microdialysis in the substantia nigra. DES (1 microM to 10 mM) co-perfused with MPP+ (2.5 mM) on day 1, produced on day 2 a higher dopamine extracellular output after perfusion of MPP+ than in control-MPP+ perfusion experiments, in which no DES was administered on day 1. Both Ringer's perfusion alone (control-Ringer) and co-perfusion of DES (10 mM) with MPP+ (2.5 mM) on day 1 produced on day 2 similar increases in dopamine extracellular output after a second MPP+ perfusion. In the control-Ringer experiment, note that the MPP+ on day 2 is the first MPP+ perfusion. Perfusion of FeCl3 (200 microM) along with MPP+ (2.5 mM) and DES (100 microM) on day 1 completely abolished on day 2 the neuroprotective effect found with MPP+ (2.5 mM) and DES (100 microM). The ability of DES to protect against MPP+ toxicity may indicate a therapeutic strategy in the treatment of diseases when iron is implicated. PMID- 9409708 TI - Acute ethanol alters calcium signals elicited by glutamate receptor agonists and K+ depolarization in cultured cerebellar Purkinje neurons. AB - The effect of acute ethanol on Ca2+ signals evoked by ionotropic (iGluR) and metabotropic (mGluR) glutamate receptor (GluR) activation and K+ depolarization was examined in cultured rat cerebellar Purkinje neurons to assess the ethanol sensitivity of these Ca2+ signaling pathways. Mature Purkinje neurons approximately 3 weeks in vitro were studied. iGluRs were activated by (RS)-alpha amino-3-hydroxyl-5 methyl-4-isoxazolepropionic acid (AMPA; 1 and 5 microM) and domoate (5 microM). mGluRs were activated by (1S,3R)-1-aminocyclopentane-1,3 dicarboxylic acid (ACPD; 300 microM) and (R,S)-3,5-dihydroxyphenylglycine (DHPG; 200 microM). These agents and K+ (150 mM) were applied from micropipettes by brief (1 s) microperfusion pulses. Ca2+ levels were monitored at 2-3 s intervals during pre- and post-stimulus periods using microscopic digital imaging and the Ca2+ sensitive dye fura-2. iGluR and mGluR agonists and K+ produced abrupt increases in intracellular Ca2+ that slowly recovered to baseline resting levels. Acute exposure to ethanol at 33 mM (150 mg%) and 66 mM (300 mg%) significantly reduced the amplitude of the Ca2+ signals to iGluR agonists and K+ with little or no effect on Ca2+ signals to mGluR agonists. In contrast, acute ethanol at 10 mM (45 mg%) had no effect on the Ca2+ signals to the iGluR agonist AMPA but significantly enhanced the Ca2+ signals to the mGluR agonist DHPG. These results show that ethanol modulates Ca2+ signaling linked to GluR activation in a receptor subtype specific manner, and suggest that Ca2+ signaling pathways linked to GluR activation and membrane depolarization may be important mechanisms by which ethanol alters the transduction of excitatory synaptic signals at glutamatergic synapses and thereby affects intercellular and intracellular communication in the CNS. PMID- 9409709 TI - Arachidonic acid inhibits uptake of amino acids and potentiates PKC effects on glutamate, but not GABA, exocytosis in isolated hippocampal nerve terminals. AB - Arachidonic acid (AA), the putative retrograde messenger in long-term potentiation, enhanced extracellular aspartate, glutamate, and GABA levels in rat hippocampus synaptosomes. Whether this effect was determined by stimulating the release and/or inhibiting the uptake of amino acids was further investigated using different experimental conditions. To approach physiological conditions, a static incubation assay was used where both release and uptake occur. Under these conditions, AA dose-dependently (10-25 microM) enhanced basal extracellular amino acid levels in a completely Ca2+-independent way. AA still exerted this effect in the presence of inhibitors of PKC or of AA metabolism. When using the superfusion release assay, in which amino acid uptake cannot occur, no potentiating effect of AA on superfusate amino acid levels was observed. Therefore, AA possibly enhances the extracellular levels of aspartate, glutamate and GABA by inhibiting the uptake of these amino acids and not their efflux. Indeed, AA reduced the Na+ dependent uptake of endogenously released amino acids, which were labelled with traces of tritiated D-aspartate and GABA. When stimulating hippocampus synaptosomes with 4-aminopyridine, AA (2 microM) potentiated the Ca2+-dependent release of glutamate, but not of GABA, synergistically with PKC activation by 4beta-phorbol-12,13-dibutyric acid. In rat hippocampus, AA exerts different presynaptic effects to regulate extracellular amino acid levels, by inhibiting carrier-mediated uptake and, for glutamate, by stimulating exocytosis. PMID- 9409710 TI - Differential effects of a benzodiazepine on synaptic transmissions in rat hippocampal neurons in vitro. AB - The effects of midazolam, one of the most popular benzodiazepines, on synaptic transmissions were compared with intracellular recordings between CA1 pyramidal cells (CA1-PCs) and dentate gyrus granule cells (DG-GCs) in rat hippocampal slices. First, we studied the effects of midazolam on orthodromically evoked spikes, membrane properties and synaptic potentials. Secondly, the effects of a GABA(A) receptor agonist, muscimol, were examined on membrane properties to determine whether or not the densities of GABA(A) receptors are different between CA1-PCs and DG-GCs. Midazolam (75 microM) markedly depressed orthodromically evoked spikes in CA1-PCs, compared with those in DG-GCs. A GABA(A) receptor antagonist, bicuculline (10 microM), almost completely antagonized the depressant effects of midazolam on spike generation in CA1-PCs, whereas it had little effect on midazolam in dentate gyrus granule cells. Midazolam produced either depolarizing or hyperpolarizing effects on resting membrane potentials (Vm) with an input resistance decrease in CA1-PCs, whereas it produced depolarized Vm in DG GCs. Midazolam significantly increased the amplitude of monosynaptic inhibitory postsynaptic potentials in CA1-PCs, whereas midazolam slightly decreased these in DG-GCs. Midazolam significantly decreased the amplitude of excitatory postsynaptic potentials both in CA1-PCs and DG-GCs. Muscimol (100 microM) produced either depolarizing or hyperpolarizing effects on Vm with an input resistance decrease in CA1-PCs, and it depolarized Vm with an input resistance decrease in DG-GCs. These results demonstrate that midazolam has differential effects on excitatory and inhibitory synaptic transmissions in hippocampal neurons. The mechanism of this difference could be partly due to the different types of GABA(A) receptors between CA1-PCs and DG-GCs. PMID- 9409711 TI - Dimethoxyphenylethylamine and tetrahydropapaverine are toxic to the nigrostriatal system. AB - We report the toxic effects of 3,4-dimethoxyphenylethylamine (DMPEA), and tetrahydropapaverine (THP) on the rat nigrostriatal system; THP is a tetrahydroisoquinoline compound which may be derived from DMPEA by conjugation of DMPEA and its oxidative metabolite, dimethoxyphenylacetaldehyde; both are potent inhibitors of mitochondrial complex I. These compounds were introduced to the unilateral caudate-putamen of male Sprague-Dawley rats over 7 days using a 200 microl mini-osmotic pump. Striatal dopamine on the injected side showed a significant decrease to 86% of the non-injected side after 16.55 micromol/7 days infusion of DMPEA, and to 73% of the non-injected side after 7.90 micromol/7 days of THP infusion; as the non-injected side dopamine also reduced in the THP injected rats, dopamine on the injected side was 55% of the saline control. Tyrosine hydroxylase (TH)-positive nigral neurons were decreased to 76% of the non-injected side after 16.55 micromol/7 days infusion of DMPEA and to 77% after 7.90 micromol/7 days of THP infusion. Dimethoxyphenyl-tetrahydroisoquinoline compounds appear to be potent nigral neurotoxins. PMID- 9409712 TI - Cholinergic, nitric oxidergic innervation in cerebral arteries of the cat. AB - Using immunoperoxidase labeling (IPL) and immunofluorescence labeling (IFL) methods, and each followed by NADPH diaphorase (NADPHd) histochemical staining in the same specimen, colocalization of choline acetyltransferase (ChAT) and NADPHd, indicative of nitric oxide synthase (NOS), in cerebral pial arteries and the sphenopalatine ganglia (SPG) of the cat was examined. In addition, retrograde axonal tracing using true blue was performed to determine if cerebral perivascular nerves containing ChAT and NADPHd originate in the SPG. Consistent results were obtained from IPL and IFL methods, indicating that the middle cerebral artery (MCA) and the circle of Willis received dense ChAT-immunoreactive (I) and NADPHd bundles and fine fibers. Almost all ChAT-I fibers and NADPHd fibers were found to be coincident in the arteries examined. A few fine fibers exhibited only NADPHd staining. In the SPG, approximately half of the ganglionic cells were both ChAT-I and NADPHd positive, while the remaining cells were positively only for NADPHd staining. One week after application of true blue on the middle cerebral arteries (MCA), the fluorescent true blue was found in the ganglionic cells of the SPG. Some of the true blue-positive cells contained both ChAT-immunoreactivity and NADPHd staining. These results provide morphological evidence indicating that all ChAT-I fibers in the MCA and the circle of Willis contain NOS, and that these fibers originate in the SPG, although not all NOS-I ganglionic cells in the SPG send fibers to pial vessels. These results also support the hypothesis that acetylcholine (ACh) and nitric oxide (NO) are synthesized and co-released in the same neurons in cerebral perivascular nerves. Based on the reported findings that NO mediates a major component of neurogenic vasodilation, and that ACh acts as a modulator, the present results demonstrate the presence of a cholinergic, nitric oxidergic innervation in cerebral arteries of the cat. PMID- 9409713 TI - Expression of beta2-thyroid hormone receptor in euthyroid and hypothyroid rat pituitary gland: an in situ hybridization and immunocytochemical study. AB - Recently, we have shown an extended distribution pattern of the TR-beta2 isoform in specific sites of rat brain which may be indicative that the localization of this receptor also confers functional specificity. The beta2 thyroid hormone receptor (TR-beta2) is by far the most abundant isoform in the pituitary, although transcripts of TR-alpha and TR-beta1 genes have been reported in developmental and adult rat pituitary gland. Using both in situ hybridization histochemistry (ISH) and immunocytochemistry, we mapped the expression of beta2 thyroid hormone receptor mRNA and protein in euthyroid and hypothyroid adult rat pituitary, particularly in relation to the thyrotrope population. TR-beta2 mRNA localization by ISH showed an anteromedial spatial distribution pattern in euthyroid rat anterior pituitary gland. This localization coincided with the immunostaining pattern for thyrotropes. TR-beta2-immunoreactive cells showed strongly positive signals in the nuclei. Hypothyroidism, induced by propylthiouracil (PTU), abolished the specific localization of TR-beta2 mRNA and upregulated the transcription of TR-beta2 mRNA in vivo and in vitro. Image analysis revealed that the optical density signals within hypothyroid rat pituitary were significantly stronger (2.6-fold) compared with euthyroid counterparts. This correlated strongly with an increased number and staining of TR-beta2 protein positive cells, demonstrating both nuclear and cytoplasmic staining. In response to thyroid hormone deficiency, there was also a marked percentage increase in the thyrotrope population from 10 to 20% of anterior pituitary cells to approximately 80%. In conclusion, these results demonstrate the specific localization of TR-beta2 to the anterior pituitary, especially to the thyrotrope population, and its regulation by thyroid hormone. Hypothyroidism leads to an upregulation of TR-beta2 mRNA and protein in the anterior pituitary, explained not only by an absolute increase in the percentage of thyrotropes but increased expression of TR-beta2 mRNA and protein per cell. These data allude to the TR-beta2 isoform playing a critical role in thyroid hormone-dependent TSH gene expression, although contributions from the other TR isoforms may still remain important. PMID- 9409714 TI - Formation of free radicals in hypoxic ischemic brain damage in the neonatal rat, assessed by an endogenous spin trap and lipid peroxidation. AB - The formation of free radicals and lipid peroxidation in the brain after hypoxic ischemia was investigated. Seven-day-old rats were subjected to unilateral (left) carotid artery ligation followed by 70 min of hypoxia with 8% oxygen at 36 degrees C. The animals were randomized into six groups as follows: control animals (no anesthesia, ligation or hypoxia) and animals decapitated at 0, 15, 30, 60 and 180 min into the reoxygenation period. Lipid peroxidation was quantified in brain homogenates using the thiobarbituric acid assay (TBA). The TBA-malondialdehyde (MDA) complex was measured with HPLC. The semi dehydroascorbate radical was measured using electron spin resonance (ESR) spectroscopy. The semi-dehydroascorbate radical levels increased more than 3-fold in the left HI hemisphere compared to the left control hemisphere 15 min posthypoxic ischemia. The amount of MDA was significantly increased in the hypoxic ischemic (HI) hemisphere ipsilateral to the carotid ligation compared with contralateral hypoxic hemisphere. The MDA level in the left HI hemisphere was also significantly elevated at 0, 15, 30 and 60 min, but not at 180 min into the reoxygenation period. Reoxygenation after hypoxic ischemia thus induced formation of semi-dehydroascorbate radicals and lipid peroxidation. PMID- 9409715 TI - Immunohistochemical localization of the neuron-specific glutamate transporter EAAC1 (EAAT3) in rat brain and spinal cord revealed by a novel monoclonal antibody. AB - Neuronal regulation of glutamate homeostasis is mediated by high-affinity sodium dependent and highly hydrophobic plasma membrane glycoproteins which maintain low levels of glutamate at central synapses. To further elucidate the molecular mechanisms that regulate glutamate metabolism and glutamate flux at central synapses, a monoclonal antibody was produced to a synthetic peptide corresponding to amino acid residues 161-177 of the deduced sequence of the human neuron specific glutamate transporter III (EAAC1). Immunoblot analysis of human and rat brain total homogenates and isolated synaptosomes from frontal cortex revealed that the antibody immunoreacted with a protein band of apparent Mr approximately 70 kDa. Deglycosylation of immunoprecipitates obtained using the monoclonal antibody yielded a protein with a lower apparent Mr (approximately 65 kDa). These results are consistent with the molecular size of the human EAAC1 predicted from the cloned cDNA. Analysis of the transfected COS-1 cells by immunocytochemistry confirmed that the monoclonal antibody is specific for the neuron-specific glutamate transporter. Immunocytochemical studies of rat cerebral cortex, hippocampus, cerebellum, substantia nigra and spinal cord revealed intense labeling of neuronal somata, dendrites, fine-caliber fibers and puncta. Double label immunofluorescence using antibody to glial fibrillary acidic protein as a marker for astrocytes demonstrated that astrocytes were not co-labeled for EAAC1. The localization of EAAC1 immunoreactivity in dendrites and particularly in cell somata suggests that this transporter may function in the regulation of other aspects of glutamate metabolism in addition to terminating the action of synaptically released glutamate at central synapses. PMID- 9409716 TI - Different receptor mechanisms mediate the effects of endotoxin and interleukin-1 on female sexual behavior. AB - Activation of the immune system by lipopolysaccharide (LPS) produces physiological, neuroendocrine and behavioral effects, some of which are mediated by cytokine production. We have previously shown that the cytokine interleukin-1 (IL-1) inhibits sexual behavior in female, but not male rats, while producing a comparable suppression of locomotion in both sexes. The present study examined the effects of LPS on sexual behavior and locomotion of male and female rats, and the involvement of IL-1 receptors in mediating the effects of IL-1 and LPS on females' behavior. Peripheral (i.p.) administration of LPS (50 or 250 microg/kg) significantly decreased sexual behavior in females, up to 6 h after administration, while it had no effect on male sexual behavior. However, locomotor activity, measured in the open-field test, was similarly reduced by LPS in both males and females. Pretreatment with the IL-1 receptor antagonist (IL 1ra) either i.p. (10 mg/kg) or intracerebroventricularly (i.c.v.) (50 microg/rat) did not prevent the inhibition of female sexual behavior and locomotion induced by either i.p. (50 microg/kg) or i.c.v. (200 or 400 ng/rat) administration of LPS, respectively. However, identical doses of IL-1ra significantly reversed the effects of IL-1beta, administered either i.p. (5 microg/kg) or i.c.v. (50 ng/rat), respectively. These results demonstrate that both LPS and IL-1beta produce marked inhibition of sexual behavior in female, but not in male rats. However, IL-1 receptors are not required for the effects of LPS on sexual behavior in female rats. PMID- 9409717 TI - Electrophysiology of excitatory and inhibitory afferents to rat histaminergic tuberomammillary nucleus neurons from hypothalamic and forebrain sites. AB - Anatomical evidence exists for projections to the tuberomammillary nucleus (TM) from the nucleus of the diagonal band of Broca (DBB) and the lateral preoptic area (LPO). The physiological effects of activating these inputs were studied by recording postsynaptic responses intracellularly from TM cells during both electrical stimulation and local nanodrop application of glutamate in horizontally cut brain slices. Electrical stimulation of the DBB, LPO and anterior lateral hypothalamic area (LH) usually evoked fast IPSPs (approximately 75% of responses) which were blocked by bicuculline or picrotoxin, suggesting GABA(A) mediation. The remaining excitatory responses evoked by stimulation of the LPO and LH were blocked by non-NMDA receptor antagonists (CNQX or NBQX) and the NMDA receptor antagonist, AP-5. Glutamate applied to the above areas induced postsynaptic responses in TM cells similar to those seen with electrical stimulation. Spontaneous firing in TM cells was suppressed by glutamate applied in the DBB. Glutamate applied in the LPO or LH evoked both inhibitory and excitatory responses. Changes in PSPs and firing rates were interpreted to result from glutamate activation of the neurons in the DBB, LPO and LH areas with inhibitory or excitatory connections to recorded TM neurons. These results support previous anatomical findings and suggest that inhibitory and excitatory synaptic control of TM activity is exerted by the DBB, LPO and LH areas. PMID- 9409718 TI - Strychnine-induced epileptiform activity in hippocampal and neocortical slice preparations: suppression by the organic calcium antagonists verapamil and flunarizine. AB - Alongside GABA, glycine is the major inhibitory transmitter in the central nervous system. Application of the glycine receptor blocker strychnine is known to evoke epileptiform phenomena. The present paper addresses the question whether postsynaptic calcium currents through L-type channels contribute to strychnine induced epileptiform field potentials (EFP). To test for this, the antiepileptic effect of the specific L-type calcium channel blocker, verapamil, in hippocampal and neocortical slices was investigated. In parallel with this, the antiepileptic efficacy of the unspecific calcium channel modulator, flunarizine, was tested with respect to pharmacotherapy of epilepsies. In both preparations, the L-type calcium channel blocker, verapamil, was able to suppress EFP. In neocortical slices, EFP were blocked in all experiments, whereas in hippocampal slices, in 3 out of 11 experiments, no complete suppression occurred. Flunarizine acted in a similar way. It is concluded that L-type calcium channels are involved in strychnine-induced epilepsy, but to a greater extent in the neocortex than in the hippocampus. PMID- 9409719 TI - Toxin I, but not 4-aminopyridine, blocks the late inhibitory component of the dorsal root reflex in an isolated preparation of rat spinal cord. AB - Bursts of spontaneous antidromic dorsal root action potentials, and evoked dorsal root reflexes (DRR), have been recorded from lumbar roots of isolated spinal cord preparations of rats weighing 70-90 g. The pattern of dorsal root activity was similar to that reported for isolated cord preparations from hamsters, but the frequency of spontaneous dorsal root activity was approximately 10 times slower in the rat. Toxin I and 4-aminopyridine (4-AP) both increased the frequency of spontaneous dorsal root activity. The threshold concentration of 4-AP was 1 microM, with EC50 at 20 microM. Insufficient Toxin I was available to reach a maximal response, but the threshold concentration for producing an increase in spontaneous activity was 0.1 microM, and the curve appeared to be parallel to that of 4-AP. The patterns of spontaneous dorsal root activity in the presence of 4-AP and Toxin I differed. In 4-AP bursts of large amplitude action potentials were followed by periods of depressed activity lasting up to 500 ms, whereas in Toxin I bursts of large amplitude action potentials caused no change in the continuously firing small amplitude action potentials. DRR evoked by stimulation of adjacent dorsal roots also showed differences in the presence of 4-AP and Toxin I. In 4-AP the excitatory phase of the reflex was followed by a period of depressed activity lasting up to 500 ms. This was was reduced or absent in the presence of Toxin I. Paired pulse stimulation confirmed the presence of inhibition in 4-AP, and its reduction in Toxin I. Examination of the pattern of spontaneous dorsal root activity following dorsal root stimulation showed strong oscillatory activity in the presence of 4-AP, but little such activity in the presence of Toxin I. It was concluded that the actions of 4-AP and Toxin I on the isolated preparation of rat spinal cord are similar in that both cause an increase in the spontaneous dorsal root firing rate, but that Toxin I also blocks the period of inhibition which follows bursts of large amplitude action potentials. PMID- 9409720 TI - Effects of pituitary adenylate cyclase-activating polypeptide (PACAP) on corticotropin-releasing hormone (CRH) gene expression in the rat hypothalamic paraventricular nucleus. AB - Pituitary adenylate cyclase-activating peptide (PACAP) is a 38-amino-acid polypeptide, first isolated from hypothalamus, which directly stimulates in vitro the production of cAMP as well as the release of several pituitary hormones, such as growth hormone and luteinizing hormone. In vivo, PACAP has been shown to stimulate ACTH release. The presence of PACAP receptors in several brain areas, including the hypothalamus, suggests that this peptide might play a role as a neurotransmitter/neuromodulator and might be involved in the regulation of hypophysiotropic neurohormones. In order to study the role of PACAP on corticotropin-releasing hormone (CRH) neuron, we have investigated the effects of intracerebroventricular (i.c.v.) and intravenous (i.v.) injections of PACAP and the potent PACAP antagonist PACAP(6-38) on CRH gene expression in the hypothalamic paraventricular nucleus (PVN) in the male rat. The levels of CRH mRNA were evaluated by quantitative in situ hybridization. The i.c.v. injection of PACAP (4 microg/kg b.wt.) produced a 22% increase in the hybridization signal, an effect which was completely prevented by the concomitant injection of the PACAP antagonist (4 microg/kg b.wt.). On the other hand, the administration of the PACAP antagonist induced by itself a 40% decrease in the amounts of CRH mRNA. The i.v. injection of the same peptides (100 microg/kg. b.wt.) produced very similar results. These data strongly suggest that PACAP is involved in the positive regulation of CRH gene expression via specific central receptors and then can play a role as a neurotransmitter/neuromodulator. The effect observed after i.v. injection of PACAP also suggests that the circulating levels of PACAP can play a role in the modulation of CRH gene expression. PACAP might then be involved in the regulation of the HPA axis by a double mechanism: stimulation of CRH gene expression at the central level and direct effect on pituitary corticotrophs. PMID- 9409721 TI - Neurofilaments are part of the high molecular weight complex containing neuronal cdc2-like kinase (nclk). AB - A cdc2-like kinase (nclk, comprised of cdk5 and its activator p25nck5a) that localizes primarily to neuronal cells has recently been identified. Although its precise physiological role remains unclear, a variety of nuclear and cytoplasmic proteins that may be targets for phosphorylation by this kinase have been suggested in various developmental and pathological states. Here we provide evidence for a functional association between nclk and neurofilament proteins: (i) brain neurofilament preparations include cdk5 and p25nck5a; (ii) nclk copurifies with neurofilament proteins using Mono-S and gel-filtration column chromatographic procedures; (iii) neurofilaments are coprecipitated with cdk5 kinase; and (iv) the addition of radiolabeled ATP to the immunoprecipitated complex results in phosphorylation of the cytoskeletal protein. These results are consistent with the formation of a functional macromolecular complex between nclk and neurofilaments in vivo and suggest a possible role for this kinase in regulating neuronal cytoskeletal networks. PMID- 9409722 TI - Repeated cocaine and stress increase dopamine clearance in the rat medial prefrontal cortex. AB - The effects of repeated footshock stress or cocaine on the kinetics of dopamine clearance in the medial prefrontal cortex (mPFC) were measured by rotating disk electrode voltammetry (RDEV). Five groups of rats were used: animals were either naive (non-handled), pre-treated with five daily saline (1 ml/kg i.p.) or cocaine (15 mg/kg i.p.) injections, or pre-treated with five daily 20-min sessions of sham shock or footshock (0.05 mA/200 ms/s). Dopamine clearance was measured after a 1-week withdrawal period. No difference in Km values was present among the treatment groups, with the mean Km value at approximately 0.5 microM for all groups. However, Vmax values were approximately 50% higher in daily sham shock-, footshock- and cocaine-pre-treated animals compared to naive rats. The increased ability to remove dopamine in these animals suggests that altered dopamine clearance may serve an adaptive mechanism in the mPFC. PMID- 9409723 TI - Quantitative measurement of protein kinase C immunoreactivity in rod bipolar cells of the goldfish retina. AB - The intensity of the immunohistochemical reaction (IIR) against the alpha species of protein kinase C (PKC) was quantified in the rod bipolar cells (RBC) of the goldfish retina using of image analysis. Retinae incubated in control Ringer solution showed similar IIR in both the soma and the axon terminal (IIR-ratio approximately 1). Activation of PKC induces the 'transport' of the enzyme to the synaptic terminal of RBC and an increase in the IIR-ratio. In the present report, the effect of retinal neurotransmitters on the IIR-ratio and the time course of PKC transport was studied. PMID- 9409724 TI - Caffeine-induced disturbances of early neurogenesis in whole mouse embryo cultures. AB - In toto mouse embryos were cultivated at embryonic day 8.5 for 26 h with 105, 310 or 620 microM caffeine; 105-310 microM correspond to concentrations transferred by the placenta of heavy caffeine consumers. Failure of neural tube closure, excessive proliferation of neuroepithelial cells and premature evagination of telencephalic vesicles were present in 50% of treated embryos. When reaching the embryonic neural tube before neuronal migration, caffeine regionally modifies the schedule and/or rate of neural cell proliferation. PMID- 9409725 TI - Effects of serotonin on synaptic and intrinsic properties of rat subicular neurons in vitro. AB - Intracellular recordings were performed to study the effects of 5-HT on membrane properties and EPSP/IPSP responses of subicular neurons in rat combined hippocampal-entorhinal cortex slices. Application of 5-HT induced in 76% of the investigated subicular cells a hyperpolarization and a reduction of membrane resistance. In bursting neurons, 5-HT caused a reduction of the depolarizing envelope underlying burst discharges and attenuated the subsequent afterhyperpolarization. While 5-HT decreased isolated AMPA/kainate and NMDA receptor-mediated responses as well as slow IPSPs, we could not find a consistent effect on isolated fast IPSPs. Since in approximately 25% of subicular neurons EPSPs and slow IPSPs were reduced without any increase of membrane conductance, we conclude that 5-HT has in addition to membrane effects also effects on synaptic currents. PMID- 9409726 TI - Inhibition of striatal energy metabolism produces cell loss in the ipsilateral substantia nigra. AB - This study examined whether damage to dopamine (DA) nerve terminals via inhibition of energy metabolism in the striatum would result in the retrograde loss of cell bodies in the substantia nigra. Infusion of 2 micromol malonate into the left striatum of rats resulted in a 67% loss of striatal DA and a 40% loss of tyrosine hydroxylase (TH)-positive neurons in the substantia nigra. No change in the number of Nissl-positive-TH-negative neurons was observed. These findings demonstrate the retrograde destruction of DA cell bodies in the substantia nigra resulting from energy impairment at their terminal projection site. PMID- 9409727 TI - Peroxynitrite-induced toxicity in cultured astrocytes. AB - The exposure of cultured astrocytes to peroxynitrite (ONOO-) for 40 min resulted in a concentration-dependent increase in the release of lactate dehydrogenase from the cells into the bathing medium over the following 24 h. Control experiments showed that the breakdown products of ONOO- contribute, to some extent, to its ability to cause cell death but that the drug vehicle (0.3 M NaOH), which increased the pH of the bathing medium to 9.4, had little effect. The cytotoxic action of ONOO- was mimicked by 3-morpholinosydnonimine (SIN-1) which liberates both nitric oxide (NO) and superoxide but not by S nitrosoglutathione which liberates only NO. SIN-1-induced cytotoxicity was reversed in a concentration-dependent manner by superoxide dismutase and attenuated by haemoglobin suggesting that the effect of SIN-1 is due, at least in part, to the formation of ONOO-. PMID- 9409728 TI - An inhibitory effect of interferon gamma on the injury-induced astrocyte proliferation in the postmitotic rat brain. AB - Influence of interferon gamma (IFNgamma) on astrocytes proliferating in response to unilateral injury of the cerebral hemisphere was investigated in 30-day-old rats. The brain injury was followed by an immediate injection of IFNgamma into the lesion cavity. On 1st or 2nd day following injury the animals were injected with [3H]thymidine and killed 4 h after the injection. The proliferating astrocytes were visualised by combination of immunocytochemical staining for glial fibrillary acidic protein (GFAP) and autoradiography. Thereafter, numbers of GFAP-immunopositive and autoradiographically-labeled astrocytes located within the region of injury were counted. On the day 1 after injury no effect of IFNgamma administration was seen. However, on day 2 the average 43% reduction of the number of proliferating astrocytes was recorded. The reduction showed no dose dependent changes. This in vivo evidence of IFNgamma-induced suppression of astrocyte proliferation was considered in relation to other determinants of astrocyte reactivity existing in the injured brain. PMID- 9409729 TI - Relative stability of carotenoid radical cations and homologue tocopheroxyl radicals. A real time kinetic study of antioxidant hierarchy. AB - Real time detection following laser flash photolysis of transient carotenoid radical cations and tocopheroxyl radicals formed in chloroform and bleaching of the carotenoids has allowed interaction between carotenoids and tocopherols to be studied. It is found that alpha-, beta-, and gamma-tocopherol reduce all the carotenoid radical cations investigated whereas the delta-tocopheroxyl radical can be reduced by lycopene and beta-carotene. Astaxanthin, canthaxanthin, and beta-apo-8'-carotenal radical cations are scavenged rapidly by all four tocopherol homologues whereas the other carotenoid radical cations react much more slowly with the tocopherols. The results allow the antioxidant hierarchy to be established: alpha-tocopherol > lycopene approximately beta-tocopherol approximately gamma-tocopherol > beta-carotene > zeaxanthin approximately delta tocopherol > lutein > echinenone >> canthaxanthin approximately beta-apo-8' carotenal > astaxanthin. PMID- 9409730 TI - Identification of potential activators of proteinase-activated receptor-2. AB - In order to identify physiological activators of proteinase-activated receptor-2 (PAR-2), a peptide chloromethane inhibitor (biotinyl-Ser-Lys-Gly-Arg-CH2Cl) based on the cleavage site for activation of PAR-2 was synthesised and tested with 12 trypsin-like serine proteinases. The second-order rate constant (ki/Ki) for the formation of the covalent proteinase-inhibitor complex varied by 2 x 10(5)-fold between the proteinases. Biotinyl-Ser-Lys-Gly-Arg-CH2Cl reacted very rapidly with trypsin, acrosin from sperm and tryptase from mast cells: the ki/Ki values with these proteinases were greater than 10(5) M(-1) x s(-1). Thus, the specificity of these proteinases matched the sequence of the activation site of PAR-2 and it can be concluded that these proteinases are potential physiological activators of PAR 2. PMID- 9409731 TI - Saposins (sap) A and C activate the degradation of galactosylceramide in living cells. AB - In loading tests using galactosylceramide which had been labelled with tritium in the ceramide moiety, living skin fibroblast lines derived from the original prosaposin-deficient patients had a markedly reduced capacity to degrade galactosylceramide. The hydrolysis of galactosylceramide could be partially restored in these cells, up to about half the normal rate, by adding pure saposin A, pure saposin C, or a mixture of these saposins to the culture medium. By contrast, saposins B and D had little effect on galactosylceramide hydrolysis in the prosaposin-deficient cells. Cells from beta-galactocerebrosidase-deficient (Krabbe) patients had a relatively high residual galactosylceramide degradation, which was similar to the rate observed for prosaposin-deficient cells in the presence of saposin A or C. An SV40-transformed fibroblast line from the original saposin C-deficient patient, where saposin A is not affected, showed normal degradation of galactosylceramide. The findings support the hypothesis, which was deduced originally from in vitro experiments, that saposins A and C are the in vivo activators of galactosylceramide degradation. Although the results with saposin C-deficient fibroblasts suggest that the presence of only saposin A allows galactosylceramide breakdown to proceed at a normal rate in fibroblasts, it remains to be determined whether saposins A and C can substitute for each other with respect to their effects on galactosylceramide metabolism in the whole organism. PMID- 9409732 TI - Oxygen dependence of photosynthetic electron transport in a bacteriochlorophyll containing rhizobium. AB - Bacteriochlorophyll-containing rhizobia, which form nitrogen-fixing nodules on the stems and roots of the legume Aeschynomene, grow photosynthetically only in the presence of oxygen or auxiliary electron acceptors. We show that, in whole cells of the Rhizobium strain BTAi 1, a single-turnover excitation flash photooxidized c-type cytochrome under aerobic but not anaerobic conditions. Light induced fluorescence yield changes show that under anaerobic conditions, the primary acceptor quinone, Q(A), is predominantly in the reduced state and so unable to accept electrons. Thus, as is the case for the aerobic photosynthetic bacterium Roseobacter denitrificans, over-reduction of Q(A) likely prohibits photosynthesis under anaerobic conditions. PMID- 9409733 TI - Identification of cDNAs encoding two G protein-coupled receptors for lysosphingolipids. AB - The structural similarity of lysosphingolipids to lysophosphatidic acid (LPA) prompted a sequence-based search for lysosphingolipid receptors using cDNA sequence of the Edg2 human LPA receptor. Two closely related G protein-coupled receptors, rat H218 and human Edg3, are highly similar to Edg2. When overexpressed in Jurkat cells, H218 and Edg3 activated serum response element driven transcriptional reporter gene in response to sphingosine 1-phosphate (S1P), dihydro-S1P and sphingosylphosphorylcholine, but not to LPA. H218 and Edg3 expressed in Xenopus oocytes conferred responsiveness to S1P and dihydro-S1P in agonist-triggered 45Ca2+ efflux. Therefore, H218 and Edg3 are functional receptors for S1P and perhaps other closely related lysosphingolipids. PMID- 9409734 TI - Inhibition of MAP kinase blocks insulin-mediated DNA synthesis and transcriptional activation of c-fos by Elk-1 in vascular smooth muscle cells. AB - Insulin-stimulated DNA synthesis, MAP kinase (MAPK) activity and c-fos expression in vascular smooth muscle cells (VSMCs) was blocked by the MAPK inhibitor PD 98059. Regulation of c-fos expression by the transcription factor Elk-1 at the serum response element (SRE) is dependent on its phosphorylation by MAPK. PD 98059 also suppressed insulin-induced Elk-1 transcriptional activity through the SRE. These data show that MAPK plays a critical role in both insulin-mediated growth and Elk-1-dependent induction of c-fos in VSMCs. PMID- 9409735 TI - Expression cloning and characterization of a renal organic anion transporter from winter flounder. AB - The cDNA coding for a renal p-aminohippurate (PAH) transporter from winter flounder (Pseudopleuronectes americanus), designated fROAT, was cloned by functional expression in Xenopus laevis oocytes. fROAT is approximately 2.8 kbp in length and encodes a protein of 562 amino acids, related to the rat renal organic anion transporter ROAT1/OAT1 and the organic cation transporters OCT1 and OCT2. In oocytes, fROAT mediated probenecid-sensitive PAH uptake, with a Km for PAH of about 20 microM, and inhibited by external glutarate (GA) (1 mM). The functional characteristics suggest that fROAT is the basolateral PAH/dicarboxylate exchanger of the flounder kidney. PMID- 9409736 TI - G alpha16 protein expression is up- and down-regulated following T-cell activation: disruption of this regulation impairs activation-induced cell responses. AB - The role of heterotrimeric G proteins in T-cell activation is poorly understood. Here we show that in normal, mature human T-cells, expression of G alpha16, the 43 kDa alpha subunit of G16, varies widely, depending on T-cell activation status. Quiescent blood lymphocytes strongly up-regulate G alpha16 after Leuco A stimulation: protein expression of G alpha16 is maximal at day 4, then decreases. Consistently, in human T-cell clones, expression of G alpha16 is high in the first week following activation and decreases rapidly within the second week. In addition, permanent disruption of regulated G alpha16 expression in Jurkat T cells by stable overexpression of 43 kDa G alpha16 inhibited Leuco A-induced interleukin-2 production, CD69 up-regulation and cell apoptosis (by 58%, 46% and 74%, respectively), suggesting that coordinate regulation of G alpha16 expression is necessary for optimal activation-induced T-cell responses, and that G alpha16 proteins may be involved in the negative regulation of TCR signalling. PMID- 9409737 TI - Ubch9 conjugates SUMO but not ubiquitin. AB - Ubiquitin conjugating enzymes participate in the thioester cascade that leads to protein ubiquitination. Although Ubc9 is homologous to E2 ubiquitin conjugating enzymes we have shown that it is unable to form a thioester with ubiquitin, but can form a thioester with the small ubiquitin-like protein SUMO. Thus Ubc9 is a SUMO conjugating enzyme rather than a ubiquitin conjugating enzyme. Transacetylation of Ubc9 by SUMO is not mediated by the E1 ubiquitin activating enzyme, but by a distinct enzymatic activity. SUMO conjugation to target proteins is mediated by a different, but parallel pathway to ubiquitination. PMID- 9409739 TI - Purification and microsequencing of hyaluronidase isozymes from human urine. AB - We recently cloned the major hyaluronidase of human plasma, which we termed HYAL1. All hyaluronidase activity could be purified from human urine on an anti HYAL1 monoclonal antibody column. However, urine contains two hyaluronidases of 57 kDa and 45 kDa, whereas plasma only contains the 57 kDa activity. Microsequencing confirmed that both urinary isozymes have N-termini identical to plasma hyaluronidase, but a second N-terminus was found in the smaller isozyme, apparently derived from the terminal 25 amino acids of HYAL1, at the C-terminus. The two polypeptides of the 45 kDa isozyme resulting from endoproteolytic cleavage of the 57 kDa isoform are presumably linked by disulfide bonds. Sperm contains two isozymes of the testicular hyaluronidase, PH-20, and the lower molecular weight isozyme is believed to be an endoproteolytically processed form of the larger protein. Analogously to PH-20, the smaller isozyme of HYAL1 is likely to be a proteolytically processed product of the larger isozyme. PMID- 9409738 TI - Evidence for a novel function of the CD40 ligand as a signalling molecule in T lymphocytes. AB - The interaction of the CD40 receptor with its ligand has been shown to be crucial for the activation of B-lymphocytes. Here, we provide evidence that the pg39 molecule/CD40 ligand (gp39/CD40L) also functions as a stimulatory molecule for T lymphocytes. Activation of T-lymphocytes via gp39/CD40L induced a strong activation of Jun-N-terminal kinase (JNK) and p38-K. Activation of these kinases correlates with a stimulation of Rac1 and inhibition of Rac1 prevents gp39/CD40L triggered JNK/p38-K activation. Further, cellular stimulation via the CD40 ligand results in tyrosine phosphorylation of cellular proteins and the activation of p56(lck). Inhibition of src-like kinases inhibits Rac1 as well as JNK/p38-K stimulation suggesting a signalling cascade from the gp39/CD40L via p56(lck) and Rac1 to JNK/p38-K. PMID- 9409740 TI - Structural organization of the bovine cathelicidin gene family and identification of a novel member. AB - Cathelicidins are a group of myeloid antimicrobial peptide precursors found in a variety of mammalian species. Transcripts of this family show a highly conserved 5' region corresponding to the 5' untranslated region, signal peptide and propiece, and diverse 3' regions encoding structurally varied C-terminal sequences that correspond to mature antimicrobial peptides after proteolytic release from the precursors. To establish the size of the bovine gene family encoding these proteins, lambda genomic clones were isolated by screening a bovine library with a probe based on the conserved cDNA region of bovine members. Restriction mapping, hybridization studies and sequence analysis identified 11 distinct genes that based on the intergenic distances of contiguous genes appear to be in close physical proximity. Among these, a novel gene encoding the precursor of a putative alpha-helical antimicrobial peptide was recognized and sequenced. The novel gene appears to be expressed in bovine bone marrow myeloid cells, spleen and testis. PMID- 9409741 TI - Association of glucose-regulated protein (grp78) with human keratin 8. AB - Keratin polypeptides 8 and 18 (K8/18) are intermediate filament proteins that are expressed in 'simple-type' epithelial cells. They associate with several proteins including the 70 kDa cytoplasmic heat shock proteins (hsp70). We identified the human 78 kDa glucose-regulated protein (grp78) as a keratin-associated protein. Keratin-grp78 association was noted after co-immunoprecipitation of K8/18 from HT29 detergent solubilized cell lysates, and appears to involve non posttranslationally modified grp78. The grp78-K8/18 association is induced by culturing cells in the presence of tunicamycin or after glucose starvation. K8/18 bound grp78 can be dissociated by Mg-ATP and the association can be reconstituted in vitro using purified grp78, then redissociated again by Mg-ATP. Binding of grp78 occurs preferentially with K8, and when reconstituted does not depend on the posttranslational modification state of K8/18. Co-incubation of K8/18 with hsp70 and grp78 shows preferential association with hsp70. Our results demonstrate a direct association of grp78 with K8 under conditions that induce grp78 expression. PMID- 9409742 TI - Developmental regulation of the plant mitochondrial matrix located HSP70 chaperone and its role in protein import. AB - Changes in the level of the mitochondrial chaperone mtHSP70 have been investigated in pea (Pisum sativum) leaf mitochondria by Western blot analysis and quantified by scanning densitometry. As pea leaves develop (from 6 days to 30 days of age) the levels of mtHSP70 decrease. Analysis of the levels of the alpha subunit of the F1ATPase show that the levels of this protein remain constant throughout the same developmental period, whereas the levels of the alternative oxidase increase. In vitro import of the alternative oxidase precursor protein into pea leaf mitochondria from day 6 to day 30 leaves and quantification by scanning densitometry indicates that protein import efficiency decreases with increasing maturity of the plant cell. Results are discussed in terms of how changing levels of the mtHSP70 chaperone, as a result of plant cell development, influence the efficiency of protein import. PMID- 9409743 TI - Glucose regulates the promoter activity of aldolase A and pyruvate kinase M2 via dephosphorylation of Sp1. AB - Proliferating cells and tumour cells maintain a high glycolytic rate even under aerobic conditions. FTO2B cells, a rat hepatoma cell line, show high activities of glycolytic enzymes. Within a culture period of 48 h the cell number increases 5-fold. Replacement of glucose by pyruvate in the culture medium lowers glycolytic enzyme activity and prevents proliferation. Transfection assays revealed that glucose deprivation dramatically decreases the transcriptional activities of the Sp1-dependent aldolase and pyruvate kinase promoters leading to reduced reporter gene expression. Sp1 binding activity is also inhibited by ocadaic acid, an inhibitor of protein phosphatase 1. Western blot analyses with nuclear extracts from FTO2B cells cultured in the presence or absence of glucose revealed differences in the phosphorylation state of Sp1. From these results we conclude that glucose increases the amount of the dephosphorylated form of Sp1 which has a higher DNA binding activity. As a consequence gene expression of the glycolytic enzymes is increased which is a prerequisite for cell proliferation. PMID- 9409744 TI - Free luciferase may acquire a more favorable conformation than ribosome associated luciferase for its activity expression. AB - A variant of firefly luciferase in which the C-terminal end was extended with 44 amino acid residues served as a model protein in this study. After transcription and translation in vitro, the enzyme activity was measured when still attached to the ribosome and when released from the ribosome by incubation with RNase A or puromycin. It was found that the C-terminally extended luciferase already had activity when linked to the ribosome, but its activity was greatly increased when released from the ribosome. These results indicate that the luciferase is folded during synthesis on the ribosome; however, some conformational adjustments occur after its release from the ribosome which are required for the full expression of its enzymatic activity. PMID- 9409745 TI - Comparison of the structure-activity relationships of nociceptin and dynorphin A using chimeric peptides. AB - The aim of the present study was to delineate the functional domains of nociceptin (noc), a neuropeptide which is structurally related to dynorphin A (dyn). The binding and biological potencies towards the nociceptin (ORL1) and dynorphin A (kappa-opioid) receptors of twenty dyn/noc and noc/dyn hybrid peptides were compared with those of the parent heptadecapeptides. Replacement of as many as eleven residues in the C-terminus of dynorphin by the corresponding nociceptin sequence has no significant effect on binding and biological activity towards the kappa-opioid receptor. In marked contrast, replacement of as few as six residues (RKLANQ) in the C-terminus of nociceptin by the corresponding dynorphin sequence (LKWDNQ) dramatically impairs both affinity and activity towards the ORL1 receptor. This clearly indicates that the two neuropeptides have different functional architectures, despite the dual structural homology of both ligands and receptors. Moreover, the recombinant peptide approach led us to identify hybrids whose sequences differ only at positions 5 and 6 and displaying opposite or no receptor selectivity. One contains the dynorphin Leu5-Arg6 sequence and prefers the kappa-opioid receptor, whereas the other comprises the nociceptin Thr5-Gly6 sequence and prefers the ORL1 receptor. A third, containing the mixed dynorphin/nociceptin Leu5-Gly6 sequence, does not discriminate between the two types of receptor. PMID- 9409746 TI - Urinary trypsin inhibitor suppresses the production of interstitial procollagenase/proMMP-1 and prostromelysin 1/proMMP-3 in human uterine cervical fibroblasts and chorionic cells. AB - The mechanisms by which urinary trypsin inhibitor (UTI) prevents preterm premature rupture of fetal membrane and premature cervical ripening were investigated. We, therefore, examined the effects of UTI on the production of matrix metalloproteinases (MMPs) which closely participate in the breakdown of extracellular matrix in cultured human uterine cervical fibroblasts and human chorionic cells. UTI suppressed specifically the production of interstitial procollagenase/proMMP-1 and prostromelysin 1/proMMP-3 from both cells in a dose dependent manner (0.32-1.28 microM). This suppression was accompanied by a decrease in steady-state levels of their mRNAs. These results indicate for the first time that UTI down-regulates the production of proMMP-1 and proMMP-3 accompanying with the decrease in the expression of their mRNAs, and therefore UTI actually participates in the maintenance of fetal membranes and/or uterine cervix by overall suppression of MMP production along with the known inhibitory actions towards serine proteinases. PMID- 9409747 TI - N,N-dimethylsphingosine 1-phosphate activates human platelets. AB - We recently reported that N,N-dimethylsphingosine 1-phosphate (DMS-1-P) can be formed from N,N-dimethylsphingosine (DMS) in activated platelets [Y. Yatomi et al., Biochem. Biophys. Res. Commun. 231 (1997) 848-851]. In this study, we synthesized, for the first time, DMS-1-P and examined the functional effects of DMS-1-P and its related sphingolipids on platelets. Although exogenous DMS was inactive, its phosphorylated derivative, DMS-1-P, induced platelet intracellular Ca2+ mobilization and shape change, but not aggregation or release reactions. Since sphingosine 1-phosphate (Sph-1-P) is structurally related to DMS-1-P and activates platelets more strongly than DMS-1-P, a competitive binding experiment for [3H]Sph-1-P was performed using DMS-1-P. DMS-1-P reduced the binding of [3H]Sph-1-P to platelets almost as much as unlabeled Sph-1-P did. These results suggest that DMS-1-P activates platelets via an interaction with a platelet surface receptor for Sph-1-P. PMID- 9409748 TI - Nitric oxide mediates NMDA-evoked [3H]GABA release from chick retina cells. AB - The stimulation of NMDA receptor increased [3H]GABA release from preloaded cultured retina cells. This effect appears to be mediated by NO production, since addition of L-NA reduces NMDA-evoked [3H]GABA release. Spermine/NO complex, an NO donor, mimics the effect produced by NMDA. The addition of zaprinast, a phosphodiesterase inhibitor, as well as 8-Br-cGMP enhances the NMDA-evoked [3H]GABA release. These results agree with the existence in chick retina cells of NO/cGMP pathways and support a role for NO in NMDA-evoked events. The activation of this receptor complex through maturative stages of the retina together with the NO-mediated increase in GABA release may account for NMDA differentiative effect in culturing retina cells. PMID- 9409749 TI - Aspirin protects endothelial cells from oxidative stress--possible synergism with vitamin E. AB - A 24-h incubation with hydrogen peroxide (0.65 mM) markedly reduced viability of cultured endothelial cells. Preincubation with aspirin (3-30 microM) protected endothelial cells from hydrogen peroxide-induced toxicity and increased viability in a concentration-dependent fashion by up to 64% of control. A similar protection was observed with D-alpha-tocopherol acetate (vitamin E, 3-30 microM). The cytoprotective effects of aspirin and vitamin E against hydrogen peroxide were overadditive suggesting different mechanisms of antioxidant action. In agreement with this, cytotoxicity induced by iron, the main catalyst of oxygen radical formation, was substantially reduced by aspirin but not vitamin E. These results show that aspirin protects endothelial cells from oxidative stress possibly via binding or chelation of free cytosolic iron. Moreover, a combination of aspirin and vitamin E might be useful for the prevention of endothelial injury in cardiovascular disease, e.g. during atherogenesis. PMID- 9409750 TI - A crystallographic approach to DNA bending: prediction of nucleosome formation by DNA triple repeats and other repetitive sequences. AB - DNA bending is due to two main factors: the inherent curvature of the sequence and its flexibility. Most methods of analysis do not allow a differentiation between these two factors. In this paper I show that the flexibility of DNA sequences can be estimated from the standard deviation of roll values determined by X-ray crystallography for each base step. As an application of this approach, the nucleosome formation ability of triple repeat sequences has been determined and shown to be in agreement with the experimental results. Local variations in twist do not appear to have any influence on nucleosome formation. PMID- 9409751 TI - Glycogenin, the primer of glycogen synthesis, binds to actin. AB - We have studied the intracellular localization of glycogenin by fusing green fluorescent protein (GFP) to the N-terminus of rabbit muscle glycogenin and expressing the chimeric protein in C2C12, COS-1 and rat hepatic cells. The fusion protein showed a nuclear and cytosolic distribution and partially co-localized with actin in the cytosol. Disruption of the actin cytoskeleton with cytochalasin D led to a change in the pattern of green fluorescence, which coincided with that observed for the remaining non-depolymerized actin. The distribution of the single point mutant K324A was completely uniform and was not affected by this drug. These findings indicate that rabbit muscle glycogenin binds to actin through the heptapeptide 321DNIKKKL327, a common motif found in other actin binding proteins, which is located at the C-terminal end of this protein, and suggest that the actin cytoskeleton plays an important role in glycogen metabolism. PMID- 9409752 TI - Doxorubicin-induced apoptosis in human T-cell leukemia is mediated by caspase-3 activation in a Fas-independent way. AB - It has recently been proposed that doxorubicin (DOX) can induce apoptosis in human T-leukemia cells via the Fas/FasL system in an autocrine/paracrine way. We show here that treatment of Jurkat cells with either anti-Fas antibodies, anthracyclin drugs or actinomycin D induces the activation of CPP32 (caspase-3) and apoptosis. However, DOX treatment did not induce the expression of membrane FasL or the release of soluble FasL and co-incubation with blocking anti-Fas antibodies prevented Fas-induced but not DOX-induced apoptosis. All the morphological and biochemical signs of apoptosis induced by anti-Fas or DOX can be prevented by Z-VAD-fmk, a general caspase inhibitor. DEVD-cho, a specific inhibitor of CPP32-like caspases which completely blocks Fas-mediated apoptosis, prevented drug-induced nuclear apoptosis but not cell death. We conclude that: (i) DOX-induced apoptosis in human T-leukemia/lymphoma is Fas-independent and (ii) caspase-3 is responsible of DOX-induced nuclear apoptosis but other Z-VAD sensitive caspases are implicated in cell death. PMID- 9409753 TI - Construction, expression and characterization of a soluble form of human endothelin-converting-enzyme-1. AB - Endothelin-converting-enzyme-1 (ECE-1) belongs to the family of zinc metallopeptidases and is responsible for generating endothelin (ET) peptides from their inactive precursors the big endothelins (bigET). The enzyme is a type II integral membrane protein consisting of a short amino-terminal cytosolic domain of 56 amino acids, a single transmembrane domain and a large putative extracellular domain containing the catalytic site. Recombinant and native ECE-1 are expressed as a dimer. We have constructed a soluble form of ECE, named sECE*, by fusing the cleavable signal peptide of pro-opiomelanocortin in frame to the complete extracellular domain of human ECE-1. Stable expression of this construct in CHO cells resulted in the secretion of a fully active enzyme. In contrast to membrane-bound ECE, sECE* was expressed as a monomer, highly glycosylated, as assessed by gel filtration and Western blot. However, recombinant sECE* converted bigET-1 with similar specific activity as ECE-1a. This activity was completely inhibited by phosphoramidon, but not by thiorphan and captopril. sECE* was active in a broad range of pH, showing an optimum of 6.6-6.8 for bigET-1. Thus, the extracellular domain alone is sufficient for conferring full ECE-1 activity, inhibitors recognition and substrate specificity. PMID- 9409754 TI - Kidney-specific expression of a novel mouse organic cation transporter-like protein. AB - Using the signal sequence trap method, we have cloned a novel 12-membrane spanning transporter-like protein, termed renal-specific transporter (RST), from the mouse kidney. RST is a 553-amino-acid protein highly homologous to recently cloned organic cation transporters, e.g. it is 30% identical to rat organic cation transporter I at the amino acid level. Northern blot analysis has revealed that the RST gene is expressed abundantly and specifically in the kidney. In situ hybridization analysis has shown that RST gene expression is restricted to the renal proximal tubule, where various organic cations such as endogenous catecholamines and choline or clinically used cationic drugs are known to be actively excreted. PMID- 9409755 TI - Conformational changes in seventeen cystine disulfide bridges of bovine serum albumin proved by Raman spectroscopy. AB - Conformational changes in cystine disulfide bridges of bovine serum albumin during acid-induced isomerization (N --> F and F --> E transitions) have been studied with Raman spectroscopy. In an X-ray crystallographic study of human serum albumin, Carter and Ho reported that all disulfide bridges of the albumin molecule are in the gauche-gauche-gauche conformation [1]. On the other hand, the solution structure of bovine serum albumin examined by Raman spectroscopy differs from its crystal structure in the conformation of some of the disulfide bridges. Two Raman bands were detected at 520 and 505 cm(-1) in the disulfide stretching mode region, suggesting that the 17 disulfide bridges in the N-form of bovine serum albumin solution take both the gauche-gauche-gauche and gauche-gauche-trans conformations. The ratio of the peak intensities at 520 and 505 cm(-1) (I505/I520) is increased from 1.6 to 2.1 and from 2.1 to 6.3 on going from the N- to the F-form and from the F- to the E-form, respectively, indicating that the gauche-gauche-trans conformation of the disulfide bridges is converted to a gauche-gauche-gauche one which is the most energetically stable form during the acid-induced isomerization. However, small amounts of gauche-gauche-trans conformation still remain even in the E-form. PMID- 9409756 TI - Characterization of room temperature induced apoptosis in HL-60. AB - We found that exposure to room temperature (RT/21 degrees C) causes apoptosis in HL-60 cells. Here we characterized RT-induced apoptosis in HL-60. After exposure to RT, apoptosis starts within 6 h and more than 80% of the cells underwent apoptosis within 20 h. All cells, however, were committed to apoptosis after 16 h and no viable cells could be recovered. The caspase-1 inhibitor (YVAD-CHO) effectively blocked apoptosis, whereas the caspase-3 inhibitor (DEVD-CHO) did not. About 20% of newly obtained early passage HL-60 cells (passage 10) also underwent apoptosis by RT treatment. These data suggest that some population in HL-60 which responds to RT with apoptosis became dominant during passaging. PMID- 9409757 TI - DDC-4, an apoptosis-associated gene, is a secreted frizzled relative. AB - The differential display method has been used in our laboratory as a coincidence analysis to isolate genes expressed in common in each of three different rat tissues undergoing physiological apoptosis: mammary gland, ovarian corpus luteum and ventral prostate. The most interesting of these isolates, DDC-4, shows a clear association with apoptosis, its expression being confined to these three organs, and only during their involution. Using DDC-4 as probe, we screened a rat ovarian cDNA library to obtain full-length isolates. One isolate, Y81 clone 40, gives rise to a protein of approximately 40 kDa with coupled in vitro transcription/translation. Sequencing of this clone indicates an open reading frame of 1044 nucleotides encoding a protein of 39.7 kDa with a putative signal sequence. This clone exhibits a high homology with the cysteine-rich domain, i.e. the ligand-binding domain, of the fizzled gene family originally defined as tissue polarity genes in Drosophila. The homology of Y81 clone 40 is most extensive with the newly described secreted frizzled relatives, the frzb subfamily. PMID- 9409758 TI - Nuclear translocation of the Y-box binding protein by ultraviolet irradiation. AB - The Y-box binding protein, YB-1, is a member of a DNA binding protein family with a structurally and functionally conserved cold shock domain. Using Western blotting and immunohistochemical methods, larger amounts of YB-1 were detected in the cytosol, particularly at the perinuclear region, than in the nucleus of human cancer cells. UV irradiation increased accumulation of YB-1 in the nucleus at 20 min and thereafter. This translocation of YB-1 into the nucleus by UV irradiation was blocked by the protein kinase inhibitor H-7, but not HA-1004. Both green fluorescent protein (GFP)-YB-1 and GFP-YB-1C with the C-terminus (248-317) of YB 1 were located mainly in the cytosol, but GFP-YB-1deltaC with a deletion at the C terminus of YB-1 was located in the nucleus. YB-1 is translocated into the nucleus by UV irradiation, possibly through a protein kinase C-mediated signal transduction pathway, and the C-terminal region of YB-1 might be important for cytoplasmic retention of YB-1. PMID- 9409759 TI - Changes in anion permeability following hypotonic challenge in rat brain endothelial cells: different responses in primary cultures and in immortalised RBE4 cells. AB - Hypotonicity-induced anion permeability changes were investigated but not detected in immortalised (RBE4) rat brain endothelial cells using iodide efflux measurements. Large, rapid increases were however observed in primary cultured cells. Both cell types were reinvestigated following culture in a common growth factor-depleted medium. Responses were still undetectable in the immortalised RBE4 cells. Reduced responses were observed in the primary cultured cells that also showed altered morphology and decreased activity of another transporter, P glycoprotein. Thus both immortalisation and different culture conditions may alter functional expression in these cells of transporters involved in hypotonicity-induced anion permeability changes. PMID- 9409760 TI - Human prion proteins expressed in Escherichia coli and purified by high-affinity column refolding. AB - An efficient method is presented for the production of intact mammalian prion proteins and partial sequences thereof. As an illustration we describe the production of polypeptides comprising residues 23-231, 81-231, 90-231 and 121-231 of the human prion protein (hPrP). Polypeptides were expressed as histidine tail fusion proteins into inclusion bodies in the cytoplasm of Escherichia coli and refolded and oxidized while N-terminally immobilized on a nickel-NTA agarose resin. This 'high-affinity column refolding' facilitates the preparation of prion proteins by preventing protein aggregation and intermolecular disulfide formation. After elution from the resin the histidine tail can be removed using thrombin without cleaving the prion protein polypeptide chain. The same protocol as used here for hPrP has been successfully applied with bovine and murine prion proteins. The protein preparations are stable for weeks at room temperature in concentrated solution and are thus suitable for detailed structural studies. Preliminary biophysical characterization of hPrP(23-231) suggests that the C terminal half of the polypeptide chain forms a well-structured globular domain, and that the N-terminal half does not form extensive regular secondary structures. PMID- 9409761 TI - Lateral self-assembly of E-cadherin directed by cooperative calcium binding. AB - We report the Ca2+ binding characteristics of recombinant Ecad12, a construct spanning the first two repeats of epithelial cadherin, and demonstrate the links between Ca2+ binding and dimer formation. Sedimentation equilibrium and dynamic light scattering experiments show that weak dimerization of Ecad12 occurs in the presence of 10 mM Ca2+ (KdP = 0.17 mM), while no appreciable dimer formation was detected in the absence of Ca2+. Ca2+-induced dimerization was also observed in electron microscopy images of Ecad12. We conclude from Ca2+ titration experiments monitored by tryptophan fluorescence and flow dialysis that dimerization does not affect the equilibrium binding constant for Ca2+. However, the value of the Hill coefficient for Ca2+ binding increases from 1.5 to 2.4 as the protein concentration increases, showing that dimer formation largely contributes to the cooperativity in Ca2+ binding. Based on these observations and previous crystallographic studies, we propose that calcium acts more likely as a geometrical aligner ensuring the proper assembly of cadherin molecules, rather than a simple adhesive. PMID- 9409762 TI - The membrane-located osmosensory kinase, EnvZ, that contains a leucine zipper like motif functions as a dimer in Escherichia coli. AB - The Escherichia coli EnvZ protein is a membrane-located osmosensor, which is a typical member of histidine kinases involved in His-Asp phosphotransfer signaling. We found that EnvZ has a leucine zipper-like motif in its presumed periplasmic domain. The functional importance of this leucine zipper-like sequence was assessed by introducing a number of appropriate amino acid substitutions. The results collectively suggest that certain leucine residues in the leucine zipper-like structure play an important role in the osmotic signal transduction mediated by EnvZ. When cysteine was substituted for the crucial leucine residues, the EnvZ dimer with disulfide bridge was detected in the cytoplasmic membrane. It was thus demonstrated that the EnvZ osmosensor exists and exerts its signaling ability as a dimer. PMID- 9409763 TI - Kinetic study of penicillin acylase from Alcaligenes faecalis. AB - Penicillin acylase from Alcaligenes faecalis has a very high affinity for both natural (benzylpenicillin, Km = 0.0042 mM) and colorimetric (6-nitro-3 phenylacetamidobenzoic acid, Km = 0.0045 mM) substrates as well as the product of their hydrolysis, phenylacetic acid (Ki = 0.016 mM). The enzyme is partially inhibited at high benzylpenicillin concentrations but the triple SES complex formed still retains 43% of the maximal catalytic activity; the affinity of benzylpenicillin for the second substrate molecule binding site is much lower (K(S)' = 54 mM) than for the first one. Phenylmethylsulfonyl fluoride was shown to be a very effective irreversible inhibitor, completely inactivating the penicillin acylase from A. faecalis in a few minutes at micromolar concentrations; this compound was used for enzyme active site titration. The absolute values of the determined kinetic parameters for enzymatic hydrolysis of 6-nitro-3-phenylacetamidobenzoic acid (k(cat) = 95 s(-1) and k(cat)/Km = 2.1 x 10(-7) M(-1) s(-1)) and benzylpenicillin (k(cat) = 54 s(-1) and k(cat)/Km = 1.3 x 10(-7) M(-1) s(-1)) by penicillin acylase from A. faecalis were shown to be highest of all the enzymes of this family that have so far been studied. PMID- 9409764 TI - cDNA cloning of the translocon associated protein beta-subunit in the chick cerebellum. AB - The 'translocon associated protein' is a tetrameric complex residing in the translocation sites, in which nascent polypeptides pass through the endoplasmic reticulum membrane. The beta subunit of this complex is a single spanning membrane protein, as deduced from cDNAs deriving from canine or human epithelial tissues. We have isolated and analysed a cDNA clone of the beta subunit from chick nervous tissue, namely cerebellum. Its deduced protein sequence is 91% homologous to both the canine and the human protein sequences, showing that the molecule is highly conserved. PMID- 9409765 TI - Suppression of the Saccharomyces cerevisiae hac1/ire15 mutation by yeast genes and human cDNAs. AB - We previously reported that the Saccharomyces cerevisiae ire15 mutation results in an inositol-auxotrophic phenotype, and that human cDNAs can suppress the ire15 mutation (Nikawa, J., 1994. A cDNA encoding the human transforming growth factor beta receptor suppresses the growth defect of a yeast mutant. Gene 149, 367 372; Nikawa, J., Nakano, H., Ohi, N., 1996b. Structural and functional conservation of human and yeast HAC1 genes which can suppress the growth defect of the Saccharomyces cerevisiae ire15 mutant. Gene 171, 107-111). Herein, we present evidence that the gene responsible for the ire15 mutation is HAC1, which encodes a transcription factor for KAR2, obtained by isolating a yeast single-copy supressor gene and by performing complementation analysis. Sequencing analysis revealed that the mutant HAC1 gene obtained from the ire15 mutant contained an AAA codon at position 50 instead of the AGA codon observed in the wild-type gene, resulting in the alteration of the aa from Arg to Lys. All human cDNAs and yeast multicopy suppressors, which had been isolated as suppressors for the ire15 mutation, were able to suppress the inositol-auxotrophic phenotype but not the defect in KAR2 induction of the hac1-disrupted strain. PMID- 9409767 TI - Cloning and characterization of CneMDR1: a Cryptococcus neoformans gene encoding a protein related to multidrug resistance proteins. AB - CneMDR1, a gene encoding a protein related to several eukaryotic multidrug resistance (MDR) proteins, was identified, cloned, and characterized from a clinical isolate of Cryptococcus neoformans (Cn) (strain M1-106). Polymerase chain reaction (PCR) amplification of a DNA region encompassing conserved motifs of other MDR-like proteins was initially used to identify and clone CneMDR1. Analysis of the corresponding cDNA revealed an open reading frame punctuated by 16 introns. CneMDR1 encoded a protein (CNEMDR1) containing 1408 amino acids (aa) with a predicted mass of approximately 152kDa. Protein structure predictions suggested the presence of two putative 6-transmembrane (TM) domains as well as two ATP-binding domains, structural characteristics typical of ATP-binding cassette (ABC) proteins. Members of this superfamily, which include MDR proteins, are frequently involved in active transport of a variety of substrates across the cell membrane. Pulsed-field gel electrophoresis revealed the presence of 12 chromosomal bands in this clinical isolate of Cn. CneMDR1 was detected by hybridization on chromosome IV. High-stringency hybridization detected only one MDR-like gene. However, a second MDR-like gene (CneMDR2) was discovered during reverse transcriptase-PCR (RT-PCR) amplification using cDNA. PMID- 9409766 TI - Gene structure and sequence comparisons of the eye lens specific protein, filensin, from rat and mouse: implications for protein classification and assembly. AB - The full length cDNA sequences of rat and mouse filensin are presented, as well as the structure of the rat filensin gene. This gene spanned 31 kb and included seven introns. The first six introns were conserved in position and phase with those found in the intermediate filament (IF) protein genes of the type II (type II keratin), type III (vimentin) and type V (lamin). The last intron of the filensin was unique. As none of the filensin intron positions coincided with those unique to type I, II or IV genes, it appears that filensin is most similar to type III genes. Comparison of the deduced amino acid sequences for rat and mouse filensin with those of cow and chick, and with other species of IF proteins, indicated the C-terminal non-alpha-helical tail domain of filensin to be one of the most divergent yet found in the vertebrate IF family. The tail domain had three conserved regions which are interrupted with two regions with lower identity. Two motifs, (1) PGDVPDGxxISKAF; and (2) KVEVVESIEKxxxxxIQTYEETxxIVET, were identified as sequences which were particularly highly conserved across species. Coassembly studies using CP49 and a physiologically derived 53 kDa-fragment of filensin showed the motif (2) was not required for filament assembly in vitro. These data strengthen the view that the C-terminal non-alpha-helical domain of filensin contributes in more than one way to filensin function in the lens. PMID- 9409768 TI - Evidence for two types of subunits in the bacterioferritin of Magnetospirillum magnetotacticum. AB - In order to investigate the role of bacterioferritin (Bfr) in the biomineralization of magnetite by microorganisms, we have cloned and sequenced the bfr genes from M. magnetotacticum. The organism has two bfr genes that overlap by one nucleotide. Both encode putative protein products of 18 kDa, the expected size for Bfr subunits, and show a strong similarity to other Bfr subunit proteins. By scanning the DNA sequence databases, we found that a limited number of other organisms, including N. gonorrhea, P. aeruginosa, and Synechocystis PCC6803, also have two bfr genes. When the sequences of a number of microbial Bfrs are compared with each other, they fall into two distinct types with the organisms mentioned above having one of each type. Differences in heme- and metal binding sites and ferroxidase activities of the two types of subunits are discussed. PMID- 9409769 TI - A GTP-binding protein of Mycoplasma hominis: a small sized homolog to the signal recognition particle receptor FtsY. AB - A protein homologous to the Escherichia coli FtsY which in turn has characteristics in common with the alpha-subunit of the eukaryotic signal recognition particle receptor (SRalpha) in the membrane of the endoplasmic reticulum, was identified in Mycoplasma hominis and its encoding DNA sequenced. The aa similarity to E. coli FtsY and B. subtilis FtsY was 38% and 51%, respectively. The protein was synthesized in E. coli, purified and shown to bind GTP. Subcellular localization studies revealed that M. hominis FtsY was associated with the cytoplasmic side of the plasma membrane. The molecular mass of M. hominis FtsY was 39.1, which was significantly smaller than FtsY from the gram- E. coli. Analysis of the primary structure showed that M. hominis FtsY had no counterpart to the N-terminal part in E. coli FtsY or mammalian SRalpha, which for the last-mentioned are known to comprise the membrane-anchoring fragment. Comparison of sequenced SRalpha homologue indicates that M. hominis together with Bacillus subtilis comprise a distinct cluster of similar small SRP receptors. PMID- 9409770 TI - Molecular characterisation of ovarian cathepsin D in the rainbow trout, Oncorhynchus mykiss. AB - In fish, cathepsin D, an aspartyl protease, is believed to mediate the processing of yolk proteins in the oocyte. Cathepsin D, therefore, is vital for the production of a viable egg. This study set out to isolate and sequence the cDNA encoding cathepsin D, and to determine the developmental expression of the message in the ovary and subsequently during embryogenesis in the rainbow trout, Oncorhynchus mykiss. The full-length trout cathepsin D cDNA is 1847 base pairs (bp) long, encoding a protein of 400 amino acids (aa). The sequence consists of a putative signal peptide of 18 aa, a prosequence extending 46 aa and a mature peptide of 336 aa. The deduced sequence of rainbow trout ovarian cathepsin D shows significant homology with cathepsin D in mammals (human; 81% aa similarity), in the chicken (80% aa similarity) and in Xenopus (74% aa similarity). Our data support the contention that the primary structure of cathepsin D is highly conserved across the vertebrate phyla, from mammals to fish. Unlike cathepsin Ds in other species, however, rainbow trout cathepsin D appears to have only one putative N-glycosylation site, rather than two. The mRNA for 'ovarian' cathepsin D was expressed in both ovarian and non-ovarian tissues (liver, muscle, spleen and testis). During the development of the ovary, the highest expression levels of cathepsin D mRNA were seen at around the onset of vitellogenesis, a time when the oocytes are starting to sequester large quantities of yolk proteins. Northern hybridisation did not detect cathepsin D mRNA in either unfertilised eggs, or in fertilised eggs until after gastrulation, indicating that there is little, if any, de novo synthesis of this message at these stages of development. However, the mRNA for cathepsin D was detectable at the eyed embryo stage, and the expression of the gene increased towards the end of embryonic development. PMID- 9409771 TI - Pattern of DNA binding of nuclear proteins to the proximal Agrobacterium rhizogenes rolC promoter is altered during somatic embryogenesis of carrot. AB - Carrot cells cultured in vitro in a medium supplemented with 2,4-D (2,4 dichlorophenoxyacetic acid) proliferate as unorganized cell clusters. Upon removal of 2,4-D from the culture medium, these cells undergo somatic embryo formation through globular, heart, and torpedo stages. Since the proximal -255 bp upstream region of the rolC gene of the Ri plasmid confers somatic embryogenesis related activation on the uidA gene in transgenic carrot cell culture, we investigated the interaction of nuclear proteins with the proximal -255bp upstream sequences to characterize the mechanism of somatic embryogenesis-related activation. Gel retardation experiments revealed that there were several different profiles of the relative levels of DNA binding activities in nuclear protein extracts from calli, PEMs (proembryogenic masses), globular embryos, and heart/torpedo embryos. The binding activity associated with a fragment (-203 bp to -92 bp) of one protein (BI) was most abundant in globular embryos. Another DNA binding protein (AII) showed the highest DNA binding activity in calli, but had low binding activities in PEMs and globular embryos. Such an altered pattern of DNA binding activities of nuclear proteins may contribute to somatic embryogenesis-related activation of the rolC promoter. Competition experiments with oligonucleotides revealed that the BI protein interacts with AT-rich sequences. PMID- 9409772 TI - Cloning and identification of the Sulfolobus solfataricus lrp gene encoding an archaeal homologue of the eubacterial leucine-responsive global transcriptional regulator Lrp. AB - The lrp gene of the extreme thermophilic archaeon Sulfolofus solfataricus, encoding a homologue of the eubacterial global leucine-responsive regulatory protein, was identified by DNA sequencing and sequence comparisons on a 6.9-kb genomic fragment cloned into Escherichia coli. The S. solfataricus Lrp subunit is a 155-aa polypeptide that bears between 24.5 and 29% sequence identity with eubacterial regulatory proteins of the Lrp/AsnC family and 30.6% and 25.8% with the archaeal homologues of respectively Methanococcus jannaschii and Pyrococcus furiosus. Transcription initiation from the strong S. solfataricus lrp promoter was analyzed by primer extension mapping. The abundance of the S. solfataricus lrp messenger strongly suggests that this protein might function in archaea as a global transcriptional regulator and genome organizer, as proposed for E. coli Lrp, rather than as a local, specific regulatory protein. Our findings suggest the presence of a eubacterial type of regulatory mechanism in archaea, a situation that is noteworthy indeed, since the transcriptional machinery of archaea is more closely related to that of eukaryotes, whereas these latter apparently do not possess a homologue of Lrp. PMID- 9409773 TI - The Orct gene of Drosophila melanogaster codes for a putative organic cation transporter with six or 12 transmembrane domains. AB - Mutations at the lemming (lmg) locus of Drosophila melanogaster cause apoptotic cell death in dividing imaginal cells. Genomic DNA flanking the P element insertion corresponding to the lmg allele lmg03424 has been cloned and found to give rise to multiple transcripts. Several cDNA clones corresponding to this genomic region were isolated and shown to differ due to alternative splicing. The complete nucleotide sequences of two of the longest cDNAs were determined and found to encode proteins with similarity to mammalian organic cation transporter (OCT) proteins. One cDNA potentially encodes a protein with six transmembrane (TM) domains, corresponding to the N-terminal half of a mammalian OCT protein, whereas the other cDNA potentially encodes a protein with 12 TM domains, corresponding to the complete mammalian OCT protein. The gene giving rise to these alternative transcripts has been named Organic cation transporter-like (Orct). The previously identified Acer gene (Taylor, C.A.M., Coates, D., Shirras, A.D., 1996. The Acer gene of Drosophila codes for an angiotensin-converting enzyme homologue. Gene 181, 191 197) appears to lie within an intron of the Orct gene. PMID- 9409774 TI - The mouse Vcs2 gene is a composite structure which evolved by gene fusion and encodes five distinct salivary mRNA species. AB - Genes of the VCS (variable coding sequence) family are characterized by an extensive evolutionary divergence in the protein-coding sequence. The VCS family has been characterized by cDNA cloning from submandibular glands in the rat, mouse and humans. At the genomic level, the sequences of two members of this family are known in the rat Rattus norvegicus: the VCSA1 gene, encoding the prohormone-like polypeptide SMR1, and the VCSB1 gene, encoding a salivary Pro rich polypeptide. No genomic data were available for the VCS genes of other species. To understand the evolution of the VCS gene family better, we have now sequenced 23 kilobases (kb) of the mouse Vcs2 gene. The Vcs2 sequence reveals numerous genomic reorganizations such as an inversion, insertions of short elements and an unusually high number of long interspersed repeated elements (LINEs), which make up 42% of this region. Interestingly, Vcs2 is composed of three different VCS-like regions. The first of these regions contains all the exons necessary to encode the previously described mouse submandibular gland polypeptide MSG2alpha. This region aligns with the entire genomic sequences of rat VCSA1 and VCSB1 genes. The two other regions align with fragments of these rat sequences. The three regions are arrayed in tandem and flanked by LINEs. In particular, the third region also contains exons that were found in mRNA species from the submandibular gland. In total, we have characterized five mRNAs from mouse submandibular glands which have in common their first exon, and are produced by alternative splicing. Vcs2 is thus a single gene that arose by the fusion of three genes (or pseudogenes) of the VCS multigene family. PMID- 9409775 TI - Rapid identification and isolation of zebrafish cDNA clones. AB - A fast and economical approach, referred to as cDNA clone tagging, was adapted to identify and isolate zebrafish cDNA clones. The basic approach was to partially sequence the coding region of size selected cDNA clones and the partial sequences were then used as tags for identifying the clones through homology search. To benefit maximally from the tagging approach, two cDNA libraries, derived from embryonic and adult fish poly(A)+ RNAs, respectively, were constructed by unidirectional cloning; conceptually, they have the potential to represent all expressed zebrafish genes. A total of 1084 clones were sequenced from the two libraries, and 511 clones were identified, based on sequence homology. These identified clones were derived from at least 261 genes, encoding 48 translational machinery proteins, 47 cytosolic proteins, 43 cytoskeletal proteins, 41 nuclear proteins, 32 membrane proteins, 22 secreted proteins, 20 mitochondrial proteins and 8 proteins with an unknown location. Of the 261 distinct cDNA clones identified, 254 were isolated for the first time in the zebrafish. These tagged cDNA clones, identified and unidentified, provide rich resources for developmental analysis as well as mapping of zebrafish genome. The long-term objective of this study is to establish a tagged zebrafish gene library that can be accessed both by hybridization screening against the plasmid DNAs and by electronic screening using the sequence information. PMID- 9409776 TI - Molecular cloning of the Drosophila melanogaster gene alpha5_dm encoding a 20S proteasome alpha-type subunit. AB - Proteasomes are large, multisubunit particles that act as the proteolytic machinery for most regulated intracellular protein breakdown in eukaryotic cells. The core proteinase of this complex, known as the 20S proteasome, is a hollow barrel-shaped structure made up of four stacked rings of seven subunits each, with the outer two rings each being made up of seven distinct alpha-type subunits, and the two inner rings composed of seven different beta-type subunits. Here we present the cloning, sequencing and genetic mapping of a Drosophila melanogaster gene, alpha5_dm, encoding one of the proteasome alpha subunits. This gene, which is homologous to the yeast PUP2 and the human Zeta genes, maps to chromosome 2 at position 54B3-5. The map positions of the previously cloned proteasome genes Pros25 and Pros29 were also determined, and found to lie at positions 87B and 57B, respectively. A search for other D. melanogaster alpha5_dm like genes encoding potential isoforms of this subunit failed to identify any closely related genes. PMID- 9409777 TI - Hox-type and non-Hox homeobox gene sequences in genomic DNA of the sea urchin Holopneustes purpurescens. AB - As a preliminary step in an analysis of Hox gene expression and radial body plan specification in sea urchin development, we amplified partial homeobox sequences in H. purpurescens by PCR using degenerate primers. The primers, HoxE and HoxF (Pendleton et al., 1993), spanned a highly conserved region of 82 nucleotides encompassing amino acids 21-47 of the homeodomain. Seven Hox-type homeobox sequences and two non-Hox homeobox sequences were identified. The seven Hox-type sequences were placed provisionally in Hox paralogous groups, one in paralogous group 3, three in paralogous groups 6-8 and three in paralogous groups 9 13. The non-Hox sequences had similarities with Xlox and Gbx homeobox genes. PMID- 9409778 TI - The first intron of the myelin proteolipid protein gene confers cell type specific expression by a transcriptional repression mechanism in non-expressing cell types. AB - Chimeric genes containing portions of the mouse myelin proteolipid protein (PLP) gene fused to the lacZ reporter gene were used to detect the effect of PLP intron 1 sequences on cell type-specific expression. A transfected fusion gene containing PLP intron 1 sequences was expressed in an oligodendrocyte cell line but not in a liver cell line, consistent with endogenous PLP gene expression. However, an analogous fusion gene missing the first intron was expressed in either oligodendrocyte or liver transfected cells. These studies suggest that transcriptional repressor element(s) located in PLP intron 1 are important in extinguishing expression in non-glial cell types and that the promoter alone functions in an indiscriminate manner. This moderately large intron (>8 kb) was sequenced to aid in future fine mapping of these cell-specific regulatory element(s). PMID- 9409779 TI - Alternative splicing of a Caenorhabditis elegans gene produces two novel inhibitory amino acid receptor subunits with identical ligand binding domains but different ion channels. AB - Two full-length cDNAs, gbr-2A and gbr-2B, encoding inhibitory amino acid receptor subunits have been amplified and cloned from Caenorhabditis elegans mRNA. The 5' 732 bp of the two cDNAs, encoding 237 amino acids, are identical. The 3' 758 bp of the gbr-2B cDNA are present within the 3' untranslated region of the gbr-2A clone. As a result, the two cDNAs are predicted to encode subunits which share a common extracellular N-terminal sequence of 237 amino acids, but different, though closely related, C-terminal sequences which include four predicted membrane-spanning regions. A search of the EMBL database revealed that the sequences of the two subunits are most closely related to the alpha-subunit of the C. elegans avermectin receptor. Northern blot analysis showed the presence of two related mRNAs of approximately 2.2 and 1.5 kb in a developmentally mixed population of C. elegans. The genomic DNA sequence confirms that both mRNAs were transcribed from the same gene, gbr-2, suggesting that the closely related 3' sequences have arisen as a result of a partial gene duplication event. We propose that C. elegans is utilising alternative splicing to generate receptor subunits with identical extracellular, ligand-binding domains but different transmembrane, channel forming domains. PMID- 9409780 TI - An ecdysteroid-responsive gene in a lobster - a potential crustacean member of the steroid hormone receptor superfamily. AB - The role of ecdysteroids in modulating exoskeletal growth during the moult cycle of Crustacea has been well described. However, little is known about the action of ecdysteroids at the level of gene transcription and regulation in Crustacea. This paper reports the cloning of an ecdysteroid responsive gene, HHR3, a potential Manduca sexta MHR3 homologue in the American lobster, Homarus americanus. Levels of HHR3 expression are up-regulated in response to in vivo injections of premoult concentrations (10(-6) M) of 20-hydroxyecdysone in the epidermal and muscle tissue of the lobster after 6 h. Maximal mRNA levels are observed after 21 h before returning to basal levels. In muscle tissue, elevated levels of HHR3 mRNA follow a time course similar to elevated actin mRNA expression in response to hormonal injection. In contrast, in eyestalk tissue, the HHR3 levels decline up to 21 h post-injection before rising to basal levels after 48 h. Eyestalk, epidermal and leg muscle tissue was extracted over the moult cycle to determine the levels of expression. In muscle, HHR3 is high during the premoult period that corresponds to the period of the moult cycle when the ecdysteroid titre is high. In the epidermis, HHR3 levels are also high during the premoult with elevated levels maintained into the postmoult period. In the eyestalk, mRNA levels of HHR3 show an opposite pattern of expression with low levels during premoult and postmoult and high levels found during the intermoult period. Our results provide novel evidence for an ecdysteroid responsive gene in a crustacean that has many similarities to MHR3 in Manduca and DHR3 in Drosophila melanogaster. This raises the question of whether a similar cascade of ecdysteroid responsive genes exist in other members of Arthropoda such as the Crustacea, as has been demonstrated in Drosophila. In addition, we provide further evidence for negative feedback regulation of ecdysteroids at the site of moult-inhibiting hormone (MIH) production in the lobster eyestalk. PMID- 9409781 TI - La autoantigen binding to a 5' cis-element of rubella virus RNA correlates with element function in vivo. AB - Rubella virus genomic RNA contains a 5' stem-loop (5'(+) SL) which is required for efficient translation and replication. The La autoantigen previously was shown to bind this RNA sequence in vitro. Results reported here demonstrate that this cellular RNA-binding protein binds to the RV 5' SL RNA with sufficient specificity for the binding to occur in the presence of excess total cellular RNA. Further, the affinity of purified La for the RV sequence is similar to its affinity for known cellular substrates. To address the functional significance of La binding, mutant forms of the RV 5'(+) SL were analysed which bind La weaker or stronger than the native form. These three forms of the RV 5' SL were incorporated into RV-luciferase constructs which expressed luciferase activity in transient transfection. The level of expression from each construct correlated with the ability of its RV sequence to bind La. The detection of La/RV RNA complexes in infected cells further supported the physiological relevance of this interaction. Possible implications of autoantigen La interaction with RV RNA for rubella virus pathology and vaccine associated adverse reactions are discussed. PMID- 9409782 TI - Hepatitis C virus NS5A protein is phosphorylated in vitro by a stably bound protein kinase from HeLa cells and by cAMP-dependent protein kinase A-alpha catalytic subunit. AB - Hepatitis C virus (HCV) has a positive-strand RNA genome that codes for a polyprotein precursor, which is processed co- and post-translationally by cellular and viral proteinases into three structural and at least six non structural (NS) proteins. The NS5A protein, expressed in mammalian cells, exists in two phosphorylated forms of 56-kDa and 58-kDa. In this study, we provide evidence for a stable association between NS5A and a protein kinase from HeLa cells and hepatocellular carcinoma (HepG2) cells by co-immunoprecipitation and by affinity to immobilized glutathione-S-transferase (GST)-NS5A fusion protein produced in E. coli. This protein kinase could phosphorylate in vitro the native NS5A on serine residues, (GST)-NS5A, histone H1, and casein as substrates. In addition, the GST-NS5A was also phosphorylated in vitro by the cAMP-dependent protein kinase A-alpha catalytic subunit. PMID- 9409783 TI - Physical map of the Clostridium difficile chromosome. AB - Clostridium difficile is a causative agent in antibiotically induced diarrhea and pseudomembraneous colitis. The ability of strains of C. difficile to cause disease depends upon the presence of two toxin genes and their corresponding proteins, designated toxin A and toxin B. Previous studies conducted in this laboratory indicated that toxigenic strains of C. difficile possess both toxin genes, whereas non-toxigenic strains do not. Likewise, the studies showed that toxigenic and non-toxigenic strains of C. difficile differ significantly in chromosomal organization by ribotype analysis. Therefore, the chromosomal organization of a reference strain was investigated. Pulsed field gel electrophoresis was utilized to generate a physical map of the chromosome of the toxigenic Clostridium difficile strain ATCC 43594. Restriction digestions of whole chromosomes with the enzymes NruI and SacII generated consistent macrofragment profiles. NruI digestion resulted in 14 discernible bands containing 16 fragments of DNA. SacII digestions resulted in 14 discernible bands containing 15 fragments of DNA. Restriction digestions with both SacII and NruI resulted in 21 discernible bands containing 31 fragments of DNA. Probing of single and double digests with an extensive set of NruI and SacII single- and double-digest bands clarified the location of individual fragments in relation to one another, resulting in a restriction map of the chromosome. PCR-generated probes of five loci of C. difficile were used to map the location of seven genes on the chromosome. Finally, the addition of all fragments from NruI, SacII and NruI/SacII digestions resulted in an approximate chromosome size of 4.4 Mb. PMID- 9409784 TI - The mUBC9 murine ubiquitin conjugating enzyme interacts with the E2A transcription factors. AB - The ubiquitin-mediated degradation of cellular proteins requires the sequential activity of E1, E2 and, in some cases, E3 enzymes. Using the yeast two-hybrid system, we have cloned 1.0- and 2.5-kb cDNAs encoding the identical murine E2, or ubiquitin conjugating enzyme by virtue of its interaction with the E2A transcription factor. This cDNA encodes the 158-amino-acid protein, mUBC9, which has considerable sequence homology to UBC9 from Saccharomyces cerevisiae and HUS5 from Schizosaccharomyces pombe and is identical to the human UBC9 protein. HUS5 is essential for DNA damage repair, whereas UBC9 is necessary for G2/M progression. The human protein has been shown to correct the UBC9 defect in yeast. Antisera raised against bacterially expressed mUBC9 fusion protein recognize a murine cellular protein of approximately 18 kDa, corresponding to the predicted mobility. Unlike E2A, the mUBC9 protein level is not regulated by serum growth factors. The activity of the apparent homologues UBC9 and HUS5 suggests that mUBC9 may be involved in the degradation of key nuclear proteins that regulate cell cycle progression. PMID- 9409785 TI - Cooperation of two PEA3/AP1 sites in uPA gene induction by TPA and FGF-2. AB - We have previously shown in NIH 3T3 fibroblasts that treatment with 12-O tetradecanoylphorbol 13-acetate (TPA) or fibroblast growth factor-2 (FGF-2) activates the Ras/Erk signaling pathway in NIH 3T3 fibroblasts, leading to the induction of the urokinase-type plasminogen activator (uPA) gene. In this study, we characterize cis-acting elements involved in this induction. DNase I hypersensitive (HS) site analysis of the uPA promoter showed that two regions were enhanced after TPA and FGF-2 treatment. One was located 2.4kb upstream of the transcription start site (-2.4kb), where a known PEA3/AP1 (AGGAAATGAGGTCAT) element is located. The other was located in a previously undefined far upstream region. Sequencing of this region revealed a similar AP1/PEA3 (GTGATTCACTTCCT) element at -6.9 kb corresponding to the HS site. Deletion analysis of the uPA promoter in transient transfection assays showed that both PEA3/AP1 elements are required for full inducibility, suggesting a synergism between the two elements. When the two sites were inserted together upstream of a minimal promoter derived from the thymidine kinase gene, expression of the reporter gene was more strongly induced by TPA and FGF-2 than with either of the two elements alone. Alone, the 6.9 element was more potent than the -2.4 element. The involvement of AP1 as well as Ets transcription factors was confirmed by examining different promoter constructs containing deletions in either the AP-1 or the PEA3 element, and by using an expression plasmid for dominant negative Ets-2. Electromobility shift analyses using specific antibodies showed that c-Jun and, JunD bind to both elements with or without induction. In addition, ATF-2 binds to the -2.4-kb element even without induction and c-Fos to the -6.9-kb element only after induction. Accordingly, overexpression of c-Fos caused induction from the -6.9-kb element, but reduced induction from the -2.4-kb element. The involvement of the Ets-2 transcription factor was shown by using expression plasmids for wild-type and dominant negative Ets-2. PMID- 9409786 TI - Genomic cloning and structural analysis of the murine polymeric receptor (pIgR) gene and promoter region. AB - The role of the polymeric receptor (pIgR) is to transport polymeric IgA across various mucosal epithelial layers. Although several mammalian pIgR cDNAs, including mouse, have been cloned, genomic structure has only been partially analyzed in the human, and neither its 5'-upstream region nor its transcriptional start site is known. We report the isolation and characterization of the murine pIgR gene that spans 32 kb and contains 11 exons. The general organization of the murine gene, including its intron/exon boundaries was similar to its human homolog; however, the second intron was 7.2 kb in the mouse vs. only 0.8 kb in humans. Primer extension and 5'-RACE independently identified the identical transcriptional initiation site. Sequence analysis of 350 base pairs in the 5' flanking region revealed several motifs, including a TATA box, and putative interferon-gamma, HNF-3beta and AP1 sites. In summary, we have isolated the murine pIgR gene and described its structure and organization. PMID- 9409787 TI - Cloning of the murine tetranectin gene and 5'-flanking region. AB - The gene encoding murine tetranectin (Tna) and its 5'-flanking region was isolated and cloned from a EMBL3 SP6/T7 genomic library. The compiled nucleotide sequence was determined by sequencing, revealing a conserved Tna structure in man and mouse. Mapping of the transcription start point (tsp) suggests that murine Tna has more than one of these. In addition, no consensus TATA-box was found uptream for the putative tsp(s) in the 5'-flanking region of the gene, indicating that the murine Tna promoter belongs to the TATA-less class of genes. The cloned murine Tna was mapped to region F1-F3 on mouse chromosome 9 by fluorescence in situ hybridization (FISH). PMID- 9409788 TI - Use of mutator cells as a means for increasing production levels of a recombinant antibody directed against Hepatitis B. AB - A mutation strategy which utilises phage display technology and the Escherichia coli mutator strains, mutD5-FIT and XL1-RED, was applied to a Hepatitis B (HepB) specific single-chain Fv (scFv) to incorporate random mutations throughout the gene. Messenger RNA from a hybridoma producing antibodies against HepB was isolated, reverse transcribed and used as template for the production of scFv. Following production of the scFv protein using an E. coli expression vector (pGC), the scFv gene was recloned into a phage display vector (pHFA). This gene construct was introduced into E. coli mutator cells and the transformed cells were used as an inoculum for liquid cultures. After five cycles of growth at 37 degrees C, each followed by dilution and re-inoculation of fresh media, recombinant phage were recovered. Nucleotide sequence analysis of the scFv gene in phage selected on HBsAg-coated magnetic beads identified amino acid substitutions which produced an increase of greater than 10-fold in apparent production levels. Competitive ELISA studies showed that the selected scFv mutants appeared to have similar affinity to HBsAg as the parent scFv. The apparent increase in production was not the result of improved surface characteristics of regions uniquely exposed in scFvs, as the sites did not correlate with the variable/constant interface of the scFv variable region normally masked in Fabs or IgGs. PMID- 9409789 TI - Open lateral retinacular lengthening compared with arthroscopic release. A prospective, randomized outcome study. AB - A prospective, randomized study was performed to compare the results of arthroscopic lateral retinacular release (Group I) with those of open lateral retinacular lengthening (Group II). The study included eighty-six patients (eighty-six knees) who had had pain in the anterior peripatellar aspect of the knee and lateral patellar tilting (rotational malalignment), as seen on Merchant tangential patellofemoral radiographs, that had not improved after participation in a structured rehabilitation program for the quadriceps and the hamstrings for a minimum of six months. The mean duration of follow-up after the operative procedures was forty-six months (range, two to six years). At the time of the latest follow-up evaluation, forty (93 per cent) of the forty-three patients in Group I and all forty-three (100 per cent) of the patients in Group II had resumed the level of athletic activity that they had engaged in before the onset of symptoms (p = 0.08, analysis of variance). With the numbers available for study, no significant difference could be detected between the groups with regard to deficits in the range of motion, atrophy (the circumference) of the thigh, operative or postoperative complications, or the need for subsequent operative procedures. Additionally, no significant difference could be detected between the groups with regard to the results of open or closed-chain testing with an isokinetic dynamometer. According to the knee-rating system of Lysholm and Gillquist as modified by Tegner and Lysholm, thirty-three knees (77 per cent) in Group I had a score of 80 points or more compared with thirty-eight knees (88 per cent) in Group II. Six knees (14 per cent) in Group I had a score of less than 70 points compared with no knees in Group II. The difference in the knee ratings between the two groups was significant (p = 0.028, analysis of variance). Although there seemed to be a definite trend toward improved function of the knee in association with a longer duration of follow-up, no significant association could be detected between the duration of follow-up and improvement in the outcome measures in either group. PMID- 9409790 TI - Regeneration of meniscal cartilage with use of a collagen scaffold. Analysis of preliminary data. AB - A collagen scaffold was designed for use as a template for the regeneration of meniscal cartilage and was tested in ten patients in an initial, Food and Drug Administration-approved, clinical feasibility trial. The goal of the study was to evaluate the implantability and safety of the scaffold as well as its ability to support tissue ingrowth. The study was based on the findings of in vitro and in vivo investigations in dogs that had demonstrated cellular ingrowth and tissue regeneration through the scaffold. Nine patients remained in the study for at least thirty-six months, and one patient voluntarily withdrew after three months for personal reasons. The collagen scaffold was found to be implantable and to be safe over the three-year period. Histologically, it supported regeneration of tissue in meniscal defects of various sizes. No adverse immunological reactions were noted on sequential serological testing. On second-look arthroscopy, performed either three or six months after implantation, gross and histological evaluation revealed newly formed tissue replacing the implant as it was resorbed. At thirty-six months, the nine patients reported a decrease in the symptoms. According to a scale that assigned 1 point for strenuous activity and 5 points for an inability to perform sports activity, the average score was 1.5 points before the injury, 3.0 points after the injury and before the operation, and 2.4 points at six months postoperatively, 2.2 points at twelve months, 2.0 points at twenty-four months, and 1.9 points at thirty-six months. According to a scale that assigned 0 points for no pain and 3 points for severe pain, the average pain score was 2.2 points preoperatively and 0.6 point thirty-six months postoperatively. One patient, who had had a repair of a bucket-handle tear of the medial meniscus and augmentation with the collagen scaffold, had retearing of the cartilage nineteen months after implantation. Another patient had debridement because of an irregular area of regeneration at the scaffold-meniscus interface twenty-one months after implantation. Magnetic resonance imaging scans demonstrated progressive maturation of the signal within the regenerated meniscus at three, six, twelve, and thirty-six months. These findings suggest that regeneration of meniscal cartilage through a collagen scaffold is possible. Additional studies are needed to determine long-term efficacy. PMID- 9409791 TI - Bone formation with use of rhBMP-2 (recombinant human bone morphogenetic protein 2). AB - We examined the effect of rhBMP-2 (recombinant human bone morphogenetic protein 2), delivered in a porous poly(DL-lactic acid) implant, on bone formation in a critical-sized defect in the radial diaphysis in rabbits. A unilateral segmental defect, twenty millimeters long, was created in the radius in ninety-six skeletally mature New Zealand White rabbits. Forty-eight rabbits were evaluated at four weeks and forty-eight, at eight weeks. Six groups were studied at each time-period. The defect was left empty in one group (control), the defect was filled with an autogenous corticocancellous bone graft in one group, and the defect was filled with a porous poly(DL-lactic acid) implant containing zero, seventeen, thirty-five, or seventy micrograms of rhBMP-2 (one group each). Radiographs of the defects were made every two weeks. The percentage of the total area of the defect that was radiopaque was determined with use of computerized radiomorphometry, and this percentage was used as a quantitative measure of the extent of new-bone formation in the defect. There were time and dose-dependent responses to rhBMP-2 for as long as four weeks; thereafter, the effects of seventeen, thirty-five, and seventy micrograms of rhBMP-2 were independent of dose and time (p < or = 0.05). The defects that had been treated with either thirty-five or seventy micrograms of rhBMP-2 had a significantly greater (p < or = 0.05) area of radiopacity than the defects that had been treated with either zero or seventeen micrograms of rhBMP-2. No significant difference could be found between the defects treated with thirty-five or seventy micrograms of rhBMP-2 and the defects filled with an autogenous graft. Healing and bone formation were examined histologically and histomorphometrically as well. At four weeks, polarized light microscopy revealed remnants of poly(DL-lactic acid) only in the defects that had been filled with an implant containing zero micrograms of rhBMP 2. At eight weeks, regardless of the dose of rhBMP-2, poly(DL-lactic acid) was not visible on histological examination. The presence of multinucleated giant cells was the hallmark of the inflammatory response elicited by poly(DL-lactic acid). At four and eight weeks, macrophages and lymphocytes were also present. The intensity of the cellular response at four weeks suggested an inverse relationship between these cells and the dose of rhBMP-2 -- that is, there appeared to be more multinucleated giant cells in defects treated with zero micrograms of rhBMP-2 than in defects treated with seventy micrograms of rhBMP-2. At eight weeks, multinucleated giant cells were rare in the defects treated with seventeen, thirty-five, or seventy micrograms of rhBMP-2. Histomorphometric data at four and eight weeks indicated that the amount of bone formation in the defects treated with seventeen, thirty-five, or seventy micrograms of rhBMP-2 was equivalent to the amount in the defects treated with an autogenous graft and was significantly less (p < or = 0.05) in the untreated defects and the defects treated with zero micrograms of rhBMP-2 (p < or = 0.05). By eight weeks, only thirty-five and seventy micrograms of rhBMP-2 had restored cortices and marrow elements. PMID- 9409792 TI - Degeneration of the accessory navicular synchondrosis presenting as rupture of the posterior tibial tendon. AB - Degeneration of the accessory navicular synchondrosis may be associated with decreased function of the posterior tibial tendon in patients who are middle-aged or older. We investigated the role of ultrasonography in differentiating between degeneration of the accessory navicular synchondrosis with separation of the accessory navicular from the navicular, which has not been previously reported to our knowledge, and a rupture of the posterior tibial tendon. We studied fourteen patients (mean age, fifty-five years; range, forty-one to seventy-two years) who had an operatively confirmed injury of the accessory navicular synchondrosis. The mean duration of follow-up was thirty-nine months (range, twenty-seven to fifty four months). Preoperative radiographs demonstrated a type-II accessory navicular (an accessory navicular with a synchondrosis) in all fourteen patients. Ultrasonography, which was performed for twelve patients, demonstrated a defect in the synchondrosis in eleven patients and a normal posterior tibial tendon in all twelve. The operative findings included incomplete separation of the synchondrosis in four of the fourteen patients, complete separation of the synchondrosis and the periosteum in eight, and avulsion of the accessory navicular in two. On the basis of our findings, we concluded that post-traumatic degeneration of an accessory navicular synchondrosis may present clinically as a variant type of avulsion or rupture of the posterior tibial tendon in this age group. Ultrasonography is useful for distinguishing between complete or partial separation through the synchondrosis and rupture or attenuation of the posterior tibial tendon. PMID- 9409793 TI - Functional bracing for rupture of the Achilles tendon. Clinical results and analysis of ground-reaction forces and temporal data. AB - Fifteen patients who had sustained a rupture of the Achilles tendon were managed non-operatively with use of a functional bracing protocol, and clinical and functional performance measures were assessed after a mean duration of follow-up of thirty-one months (range, twenty-four to forty-five months). An age and gender matched group of fifteen subjects was assessed to provide normative data for the comparison of side-to-side differences. Numerical scores were generated on the basis of subjective responses to a questionnaire, clinical measurements of the range of motion of the ankle and the circumference of the calf, and the results of the Thompson squeeze test and a single-limb heel-rise test. A 100-point scoring system was used to categorize the outcome as excellent, good, fair, or poor. In addition, ground-reaction forces and temporal data were assessed during functional dynamic activities that included walking, a single-limb power hop, and a thirty-second single-limb heel-rise endurance test. The result was graded as excellent for three patients, good for nine, fair for two, and poor for one. An increase in passive dorsiflexion of the treated ankle was the only clinical measure that was significantly different between the groups (p = 0.02). This increase in dorsiflexion was positively correlated with vertical force output between the mid-stance and terminal-stance phases of gait (r = 0.40, p = 0.05). With the numbers available, we could detect no significant differences between the groups with regard to the kinetic or temporal variables that were measured during functional dynamic activities. Patients who generated less peak vertical force and vertical height during the single-limb power-hop test tended to have poorer clinical scores. We believe that non-operative functional bracing may prove to be a viable alternative to operative intervention or use of a plaster cast for the treatment of acute ruptures of the Achilles tendon. The goals of treatment are to prevent the musculoskeletal changes that are associated with immobilization, to reduce the time needed for rehabilitation, and to facilitate an early return to work and to preinjury activities. PMID- 9409794 TI - The effect of fibular malreduction on contact pressures in an ankle fracture malunion model. AB - Nine fresh-frozen cadaveric specimens were disarticulated through the knee, and the soft tissues, except for the interosseous ligaments and interosseous membrane, were removed to the level of the ankle. The subtalar joint was secured with screws in neutral position (approximately 5 degrees of valgus). Contact pressures in the tibiotalar joint were measured with use of low-grade pressure sensitive film, which was placed through an anterior capsulotomy. For each measurement, 700 newtons of load was applied to the specimen for one minute. The film imprints were scanned, and the contact pressures were quantitated in nine equal quadrants over the talar dome. A fracture-displacement device was secured to the distal end of the fibula; the device allowed for individual or combined displacements consisting of shortening, lateral shift, and external rotation of the fibula. The ankle was maintained in neutral flexion. The ligamentous injury associated with a pronation-lateral rotation fracture of the ankle was simulated by dividing the deep fibers of the deltoid ligament, the anterior-inferior tibiofibular ligament, and the interosseous membrane to a point that was an average of fifty-three millimeters proximal to the ankle joint. Baseline contact area and contact pressure in the joint were determined, followed by measurements after two, four, and six millimeters of shortening of the fibula; after two, four, and six millimeters of lateral shift of the fibula; and after 5, 10, and 15 degrees of external rotation of the fibula. The three types of displacement were tested individually as well as in combination. The simulated deformities were found to cause a shift of the contact pressure to the mid-lateral and posterolateral quadrants of the talar dome, with pressures as high as 4.1 megapascals. A corresponding decrease in the contact pressures was noted in the medial quadrants of the talar dome. The highest pressures were recorded for maximum shortening of the fibula, the combination of maximum shortening and lateral shift, the combination of maximum shortening and external rotation, and the combination of maximum shortening, lateral shift, and external rotation. In general, increases in each displacement variable corresponded to increasing contact pressures. PMID- 9409795 TI - Corrective osteotomy for malunited, volarly displaced fractures of the distal end of the radius. AB - Twenty-five patients who had had an opening-wedge osteotomy for the treatment of a malunited, volarly displaced fracture of the distal end of the radius were studied retrospectively. The indications for the operation were pain and functional limitations rather than the degree of anatomical deformity. Fifteen patients were men and ten were women; their average age was forty-six years (range, twenty-one to eighty-four years). Preoperative radiographs revealed an average ulnar inclination of 14 degrees, an average ulnar variance of five millimeters, and an average volar inclination of 24 degrees. Extension of the wrist averaged 25 degrees; flexion of the wrist, 53 degrees; supination of the forearm, 41 degrees; and pronation of the forearm, 64 degrees. The average grip strength was a force of seventeen kilograms compared with a force of forty kilograms in the contralateral hand. At an average of sixty-one months (range, eighteen to 114 months) after the osteotomy, supination of the forearm had improved to an average of 69 degrees and pronation had improved to an average of 75 degrees (p < 0.05 for both). Extension of the wrist had improved to an average of 55 degrees, and grip strength had improved to a force of thirty kilograms (p < 0.05 for both). Volar inclination averaged 5 degrees; ulnar variance, zero millimeters; and ulnar inclination, 22 degrees. A reoperation was performed in eleven patients. Seven patients had removal of the hardware only, two had a procedure involving the distal radioulnar joint, one had a procedure because the site of the osteotomy had not healed, and one had a median-nerve release. The functional result was rated as very good in ten patients, good in eight, fair in three, and poor in four. PMID- 9409796 TI - Total hip arthroplasty performed with insertion of the femoral component with cement and the acetabular component without cement. Ten to thirteen-year results. AB - Fifty-two consecutive primary total hip arthroplasties were performed in forty seven unselected patients by one surgeon. The prosthesis included a hemispherical porous-coated acetabular component, inserted without cement and with the use of screws through three peripheral flanges, and a femoral component, inserted with a so-called second-generation cementing technique. No patient was lost to radiographic follow-up, and the clinical result was known for all patients. The average age at the time of the index operation was fifty-seven years (range, twenty-nine to seventy-nine years). Four patients (four hips) who died were last examined less than ten years postoperatively (the minimum follow-up period for this study) and one hip was revised, leaving forty-seven non-revised hips in forty-two surviving patients who were followed for at least ten years. The duration of clinical follow-up of these forty-two patients averaged 12.3 years (range, 10.8 to 13.3 years), and the duration of radiographic follow-up averaged 12.1 years (range, 10.0 to 13.0 years). One (2 per cent) of the original fifty two hips was revised for late recurrent dislocation, without loosening, 9.7 years after the index arthroplasty. The rate of dislocation was relatively high (13 per cent; seven hips), and we believed it to be related to the shallow-chamfer acetabular design combined with the small femoral head. At the time of the latest follow-up, no femoral component was loose. One (2 per cent) of the fifty-two acetabular components was loose according to radiographic criteria, but the hip functioned well (Harris hip score, 94 points) 12.4 years after the index arthroplasty. Pelvic osteolysis developed in one hip (2 per cent); femoral osteolysis, in eight (15 per cent); and distal femoral osteolysis, in three (6 per cent). The average Harris hip score for the forty-seven non-revised hips increased from 48 points (range, 26 to 63 points) preoperatively to 89 points (range, 67 to 100 points) at the time of the most recent follow-up. Forty (85 per cent) of the forty-seven hips had a good or excellent result, whereas five (11 per cent) had a fair result (score, 74 to 79 points) and two (4 per cent) had a poor result (score, 67 and 69 points). The hybrid primary total hip arthroplasty resulted in very good clinical function at ten to thirteen years, although the rate of dislocation was high. PMID- 9409797 TI - Impaction bone-grafting before insertion of a femoral stem with cement in revision total hip arthroplasty. A minimum two-year follow-up study. AB - Impaction bone-grafting was performed before insertion of a collarless, polished, tapered femoral stem with cement in thirty-four revision total hip arthroplasties (thirty-four patients) that were done because of aseptic loosening. The average duration of follow-up was thirty months (range, twenty-four to forty-two months). The operation was the initial revision in twenty-eight patients (82 per cent). Twenty-two patients (65 per cent) also had revision of the acetabular component. Complications included four intraoperative and two postoperative fractures of the femur as well as one dislocation (at one month). Two patients (6 per cent) needed a repeat revision of the femoral stem because of aseptic loosening at twenty-six and thirty-six months postoperatively. Both of these patients had an associated fracture of the femur (one was intraoperative, and the other was postoperative). Subsidence was common (thirteen patients; 38 per cent) and averaged 10.1 millimeters (range, four to thirty-one millimeters). Although the study group was relatively small, with the numbers available subsidence was not found to be associated with the preoperative or postoperative hip score, segmental or cavitary femoral defects, femoral ectasia, intraoperative fracture of the femur, strut-grafting, trochanteric osteotomy, or varus position of the femoral component. Incorporation of the allograft into the trabecular bone and secondary remodeling were noted radiographically in thirty-two (94 per cent) and fourteen (41 per cent) of the patients, respectively, often within one year. Although the duration of follow-up was relatively short, no localized resorption of the allograft occurred and cortical repair was noted in one patient at three years. At the most recent follow-up evaluation, the Harris hip scores had improved from a preoperative average of 51 points (range, 32 to 90 points) to an average of 87 points (range, 65 to 100 points) and twenty-eight patients (82 per cent) had no or only slight pain. Despite the satisfactory early clinical results, we remain concerned about the high rate of fracture of the femur and the rate and extent of subsidence of the femoral component. On the basis of the worrisome findings after this two-year period, we recommend that impaction bone-grafting be used only when proximal femoral osteopenia is so severe that stability cannot be obtained with insertion of a long-stemmed femoral component without cement. In that setting, impaction bone-grafting may be considered instead of implantation of a massive proximal femoral allograft in combination with insertion of a femoral component with cement. PMID- 9409798 TI - Fatigue fracture of a forged cobalt-chromium-molybdenum femoral component inserted with cement. A report of ten cases. AB - Ten patients who had had a total hip replacement with a forged cobalt-chromium molybdenum femoral prosthesis (Precoat or Precoat Plus) inserted with cement were seen with a fatigue fracture of the stem an average of fifty months (range, nineteen to seventy-four months) postoperatively. The average age of the patients was sixty-one years (range, forty-three to seventy-three years), and the average weight was ninety-six kilograms (range, seventy to 130 kilograms). Eight patients had had a primary total hip replacement, and two had had a revision; all of the acetabular components had been inserted without cement. Radiographs that had been made before the fracture were available for four of the eight hips that had had a primary replacement; all four had radiographic evidence of debonding of the cement mantle from the proximal end of the stem. This probably caused exaggerated cantilever bending stresses on the proximal aspect of the stem as the distal end of the stem was well fixed. The radiographs of both hips that had had a revision demonstrated a non-union of the greater trochanter, which had resulted in separation at the cement-bone interface at the proximal portion of the femur before the fracture. Scanning electron micrographs of five of the ten fractured prostheses demonstrated a fatigue fracture that began near the anterolateral corner of the prosthesis, through characters that had been etched on the implant with a laser. Metallurgical analysis indicated subsurface voids or inclusions, or both, immediately under the region that had been etched. This finding is consistent with thermal changes to the microstructure of the alloy that probably caused a focal reduction in the material strength. A high proportion (seven) of the ten stems had a poor cement mantle. Also, of the seven small stems that were used, six had been implanted in patients who weighed more than eighty kilograms, so there was relative undersizing of the prostheses. Early debonding of the proximal end of a Precoat femoral prosthesis from the cement mantle may occur as a result of a thin cement mantle, leading to loosening and possibly to early fatigue fracture of the stem if the distal portion of the stem remains solidly fixed in the distal portion of the cement column. On the basis of our experience, we recommend that patients who have radiographic evidence of a debonded Precoat femoral component should be informed of the risk of fatigue fracture of the stem and be followed closely even though there may be no symptoms of loosening of the femoral component. PMID- 9409799 TI - Osteosarcoma associated with osteochondritis dissecans. A case report and review of the literature. PMID- 9409800 TI - Subacromial impingement syndrome. PMID- 9409801 TI - Current concepts review. Tuberculosis of bones and joints. PMID- 9409802 TI - Porous-coated acetabular components with screw fixation. Five to ten-year results. PMID- 9409803 TI - Current concepts review. Managed care: form, function, and evolution. PMID- 9409804 TI - Abrasive three-body wear of polyethylene caused by broken multifilament cables of a total hip prosthesis. A report of three cases. PMID- 9409805 TI - Operative treatment of fractures of the tibial plafond. A randomized, prospective study. PMID- 9409806 TI - Reflex sympathetic dystrophy and pain dysfunction in the lower extremity. PMID- 9409807 TI - Profilins as regulators of actin dynamics. PMID- 9409808 TI - Characterization of beta-amyloid peptide precursor processing by the yeast Yap3 and Mkc7 proteases. AB - Two proteases, denoted beta- and gamma-secretase, process the beta-amyloid peptide precursor (APP) to yield the Abeta peptides involved in Alzheimer's disease. A third protein, alpha-secretase, cleaves APP near the middle of the Abeta sequence and thus prevents Abeta formation. These enzymes have defied identification. Because of its similarity to the systems of mammalian cells the yeast secretory system has provided important clues for finding mammalian processing enzymes. When expressed in Saccharomyces cerevisiae APP is processed by enzymes that possess the specificity of the alpha-secretases of multicellular organisms. APP processing by alpha-secretases occurred in sec1 and sec7 mutants, in which transport to the cell surface or to the vacuole is blocked, but not in sec17 or sec18 mutants, in which transport from the endoplasmic reticulum to the Golgi is blocked. Neutralization of the vacuole by NH4Cl did not block alpha secretase action. The time course of processing of a pro-alpha-factor leader-APP chimera showed that processing by Kex2 protease, a Golgi protease that removes the leader, preceded processing by alpha-secretase. Deletions of the genes encoding the GPI-linked aspartyl proteases Yap3 and Mkc7 decreased alpha secretase activity by 56 and 29%, respectively; whereas, the double deletion decreased the activity by 86%. An altered form of APP-695, in which glutamine replaced Lys-612 at the cleavage site, is cleaved by Yap3 at 5% the rate of the wild-type APP. Mkc7 protease cleaved APP (K612Q) at about 20% the rate of wild type APP. The simplest interpretation of these results is that Yap3 and Mkc7 proteases are alpha-secretases which act on APP in the late Golgi. They suggest that GPI-linked aspartyl proteases should be investigated as candidate secretases in mammalian tissues. PMID- 9409809 TI - Cholesterol derivative of poly(ethylene glycol) inhibits clathrin-independent, but not clathrin-dependent endocytosis. AB - The effect of poly(ethylene glycol) cholesteryl ethers (PEG(n)-Chols) with two different numbers of units (n = 50 and 200) in the hydrophilic PEG moiety on cellular endocytic activity was studied on HT-1080 cells. The amphipathic molecules were soluble in aqueous solution. When fluorescein derivatives of PEG Chols (one fluorescein at the distal end of PEG) were incubated with the cells in culture, the cellular fluorescence was localized at the plasma membrane level and in intracellular vesicles. Fluorescence quantification indicated that for the same external concentration, twice more FPEG(50)-Chol than FPEG(200)-Chol was associated with the cells under the same conditions. Regardless of the length of PEG moiety, PEG-Chols' interaction with cells reduced the endocytic internalization of a fluid phase marker, horseradish peroxidase (HRP) depending on the cell-associated amount. In contrast, internalization of 125I-labeled epidermal growth factor (EGF) through receptor-mediated endocytosis did not change upon incubation with PEG(50)-Chol. The effect of PEG(200)-Chol was also small, since EGF internalization showed a reduction of 10-20%, while at the same concentration as much as 80% of HRP uptake was inhibited. PEG(50)-Chol did not influence the internalization of a larger ligand, 125I-transferrin (Tfn). However, in the presence of PEG(200)-Chol, the uptake of 125I-Tfn decreased remarkably, and yet, PEG(200)-Chol has no influence on the binding and internalization of a monoclonal antibody directed toward the ectodomain of the Tfn-receptor. These results suggested that incorporation of PEG-Chols in the outer monolayer of the plasma membrane specifically inhibited clathrin independent, but not clathrin-dependent endocytosis. PMID- 9409810 TI - Differentiation-dependent expression of obese (ob) gene by preadipocytes and adipocytes in primary cultures of porcine stromal-vascular cells. AB - The relationship between obese (ob) gene expression and preadipocyte differentiation was examined in primary cultures of porcine stromal-vascular (S V) cells by Northern-blot analysis using a pig ob cDNA probe. Isolated adipocytes expressed high levels of ob gene, but S-V cells did not express the ob gene. Cultures were seeded with fetal bovine serum (FBS) plus dexamethasone (Dex) for 3 days followed by ITS (insulin 5 microg/ml, transferrin 5 microg/ml, and selenium 5 ng/ml) treatment for 6 days. Detectable levels of ob mRNA first appeared at day 1 with very low activity of glycerol phosphate dehydrogenase (GPDH). Levels of ob mRNA increased in parallel with preadipocyte number or GPDH activity at the later times in cultures. The depletion of preadipocytes by complement-mediated cytotoxicity at day 3 of culture resulted in markedly decreased ob mRNA expression. Immunocytochemical analysis showed that ob protein was localized in the cytosol of preadipocytes and adipocytes. These data indicated that the ob gene is expressed by preadipocytes and ob gene expression may be correlated with preadipocyte recruitment as well as fat cell size. PMID- 9409811 TI - Reduced glutathione prevents nitric oxide-induced apoptosis in vascular smooth muscle cells. AB - The control of medial and neointimal growth, in which vascular smooth muscle (VSM) plays a central role, is most important to the development of hypertension and atherosclerosis, respectively. Growth of vascular smooth muscle cells is regulated by a number of factors, including the vasodilator nitric oxide (NO). In addition, NO modulates intracellular thiol redox states and the thiol redox state of the cell influences NO production. We, therefore, examined the nature of the effect of NO on growth of VSM cells and its modulation by cellular glutathione content. Here, we report that NO, either generated by NO donors or synthesized by iNOS in VSM cells, inhibited DNA synthesis and induced apoptosis in this cell type. NO-induced apoptosis was associated with a significant decrease in the intracellular concentration of reduced glutathione and with an increase in the level of the tumor suppressor gene p53 mRNA. Moreover, addition of glutathione monoethylester to the culture restored the level of reduced glutathione in VSM cells, and prevented the NO-induced increase in p53 expression and programmed cell death. Our findings suggest a role for reduced glutathione in protecting VSM cells exposed to NO from apoptosis. PMID- 9409812 TI - Endothelin association with cultured rat hepatic endothelial cells: functional characterization. AB - Endothelin is a potent vasoactive peptide whose concentration increases in a number of pathophysiological states. In the intact animal, the liver is known to sequester approximately 12% of an injected bolus of [125I]endothelin-1 ([125I]ET 1). Endothelial cells (ECs) isolated from rat liver were maintained in culture in order to examine their role in ET sequestration. LECs were shown to express predominantly ET(B) receptors both by association assays and by Northern blot analysis. In these cells the reaction between [125I]ET-1 and its receptor was essentially irreversible. Ligand binding experiments performed at 4 degrees C showed that LECs in early culture (approximately 3 h) had 4.3 +/- 0.8 fmol of ET receptors per 10(6) cells; this number fell progressively to < or = 1 fmol/10(6) cells during 24 h of culture. The decrease in receptor numbers could be blocked by maintaining the cells at 4 degrees C. Northern blot analysis showed that relative to freshly isolated cells, mRNA for the ET(B) receptor decreased 4-fold in early culture, and recovered somewhat at 24 h. At 37 degrees C [125I]ET-1 bound by the cells was rapidly internalized, with concomitant down-regulation of ET receptors. Recovery of down-regulated ET receptors was sensitive to cycloheximide, making short-term receptor recycling unlikely. Metabolism of [125I]ET-1 was low at short (< 4 h) exposure times, and at 24 h showed a concentration dependence similar to that of ligand association, suggesting that ET-1 metabolism primarily was intracellular. ET stimulation of Kupffer cells and other hepatic cell types is known to activate phosphoinositide signaling, but no such activation was seen in LECs. Moreover, ET did not appear to stimulate protein tyrosine kinase activity in LECs. While hepatic LECs may lack some of the ET-dependent responses seen in other cell types, they likely contribute substantially to the liver's previously reported ability to sequester systemically administered ET. PMID- 9409813 TI - Follicle-stimulating hormone regulation on its receptor messenger ribonucleic acid levels in cultured rat granulosa cells. AB - Our studies using immature rat granulosa cells cultured in serum-free medium on collagen-coated dishes indicated that FSH receptor mRNA levels do not change for at least 4 days of culture in the absence of hormone treatment. Addition of FSH (30 ng ml[-1]) led to a reduction of FSH receptor mRNA for a short time (6 h), followed by an increase in FSH receptor mRNA levels that reached maximum of around 200% of the initial level within 2-3 days after the addition of FSH. Following the addition of 10 nM PMA, FSH receptor mRNA levels were decreased to 50% of the pretreatment levels. During prolonged exposure to PMA, gradual recovery of the FSH receptor mRNA level was observed, and it was significantly higher than the control level at 48 h. The inactive phorbol ester 4 alpha-phorbol 12,13-didecanoate did not depress FSH receptor mRNA levels. Downregulation of the FSH receptor mRNA was detectable at a PMA concentration of 1 nM. The two predominant FSH receptor mRNA transcripts, ca. 5.5 and 2.4 kb, respectively, appeared to be equally affected by SH and PMA treatments. To examine the role of PKC mediation of the effect of FSH on FSH receptor mRNA levels, granulosa cells were treated with the PKC inhibitor, H-7, and FSH. Although, FSH receptor mRNA levels decreased to 50% of control in the cells treated with FSH alone, the addition of H-7 (0.1 nM) caused no decline in FSH receptor mRNA levels relative to the control in the cells treated with FSH. On the other hand, inhibition of FSH receptor mRNA by FSH was partially suppressed by the PKC-selective inhibitor bisindolylmaleimide. The mRNA turnover experiments showed that the half-life of FSH receptor transcripts was unaffected by PMA exposure. PMID- 9409815 TI - Seventy years ago: mutation becomes experimental. PMID- 9409814 TI - Differential activity of bcl-2 and ICE enzyme family protease inhibitors on Fas and puromycin-induced apoptosis of glioma cells. AB - Fas ligand is a potent inducer of apoptosis in human glioma cells by the Fas/Fas ligand pathway. With comparable efficiency, metalloprotease inhibitors including puromycin and bestatin induce apoptosis in glioma cells. To evaluate the involvement of potential components involved in Fas ligand- and metalloprotease inhibitor-induced apoptosis, we investigated the effect of anti human Fas antibody, soluble Fas ligand and puromycin on cultures of human malignant glioma cell lines (LN-18, LN-229, T98G). Stimulation with Fas ligand lead to apoptotic cell death within 16 h. Costimulation with the translational inhibitor cycloheximide and the transcription blocker actinomycin D did not reduce Fas ligand toxicity. In contrast, apoptosis induced by puromycin was blocked by cycloheximide and decreased by subtoxic doses of actinomycin D in all three gliomas. Whereas inhibition of caspase activity with the general inhibitor zVAD fmk resulted in a complete block of Fas ligand-induced cell death, puromycin mediated apoptosis was found to be unaffected by zVAD-fmk as well as by more specific inhibitors for caspase-1 (Interleukin-1 beta converting enzyme) and caspase-3 (CPP32/Yama). Other prominent components involved in many apoptotic pathways as bcl-2 and reactive oxygen intermediates were also examined. Bcl-2 which protects glioma cells from Fas ligand-induced cell death, was shown to have only a small protective effect on puromycin-induced apoptosis. The tested radical scavengers did not reduce Fas- or puromycin-mediated killing of human glioma cells. PMID- 9409816 TI - Exceptional convergent evolution in a virus. AB - Replicate lineages of the bacteriophage phiX 174 adapted to growth at high temperature on either of two hosts exhibited high rates of identical, independent substitutions. Typically, a dozen or more substitutions accumulated in the 5.4 kilobase genome during propagation. Across the entire data set of nine lineages, 119 independent substitutions occurred at 68 nucleotide sites. Over half of these substitutions, accounting for one third of the sites, were identical with substitutions in other lineages. Some convergent substitutions were specific to the host used for phage propagation, but others occurred across both hosts. Continued adaptation of an evolved phage at high temperature, but on the other host, led to additional changes that included reversions of previous substitutions. Phylogenetic reconstruction using the complete genome sequence not only failed to recover the correct evolutionary history because of these convergent changes, but the true history was rejected as being a significantly inferior fit to the data. Replicate lineages subjected to similar environmental challenges showed similar rates of substitution and similar rates of fitness improvement across corresponding times of adaptation. Substitution rates and fitness improvements were higher during the initial period of adaptation than during a later period, except when the host was changed. PMID- 9409817 TI - Size and sequence polymorphism in the isocitrate dehydrogenase kinase/phosphatase gene (aceK) and flanking regions in Salmonella enterica and Escherichia coli. AB - The sequence of aceK, which codes for the regulatory catalytic enzyme isocitrate dehydrogenase kinase/phosphatase (IDH K/P), and sequences of the 5' flanking region and part or all of the 3' flanking region were determined for 32 strains of Salmonella enterica and Escherichia coli. In E. coli, the aceK gene was 1734 bp long in 13 strains, but in three strains it was 12 bp shorter and the stop codon was TAA rather than TGA. Strains with the shorter aceK lacked an open reading frame (f728) downstream between aceK and iclR that was present, in variable length, in the other strains. Among the 72 ECOR strains, the truncated aceK gene was present in all isolates of the B2 group and half of those of the D group. Other variant conditions included the presence of IS1 elements in two strains and large deletions in two strains. The aceK-aceA intergenic region varied in length from 48 to 280 bp in E. coli, depending largely on the number of repetitive extragenic palindromic (REP) sequences present. Among the ECOR strains, the number of REP elements showed a high degree of phylogenetic association, and sequencing of the region in the ECOR strains permitted partial reconstruction of its evolutionary history. In S. entica, the normal length of aceK was 1752 bp, but three other length variants, ranging from 1746 to 1785 bp, were represented in five of the 16 strains examined. The flanking intergenic regions showed relatively minor variation in length and sequence. The occurrence of several nonrandom patterns of distribution of polymorphic synonymous nucleotide sites indicated that intragenic recombination of horizontally exchanged DNA has contributed to the generation of allelic diversity at the aceK locus in both species. PMID- 9409818 TI - Genetic analysis of mutations affecting pckA regulation in Rhizobium (Sinorhizobium) meliloti. AB - The enzyme phosphoenolpyruvate carboxykinase (Pck) catalyzes the first step in the gluconeogenic pathway in most organisms. We are examining the genetic regulation of the gene encoding Pck, pckA, in Rhizobium (now Sinorhizobium) meliloti. This bacterium forms N2-fixing root nodules on alfalfa, and the major energy sources supplied to the bacteria within these nodules are C4-dicarboxylic acids such as malate and succinate. R. meliloti cells growing in glucose minimal medium show very low pckA expression whereas addition of succinate to this medium results in a rapid induction of pckA transcription. We identified spontaneous mutations (rpk) that alter the regulation of pckA expression such that pckA is expressed in media containing the non-inducing carbon sources lactose and glucose. Genetic and phenotypic analysis allowed us to differentiate at least four rpk mutant classes that map to different locations on the R. meliloti chromosome. The wild-type locus corresponding to one of these rpk loci was cloned by complementation, and two Tn5 insertions within the insert DNA that no longer complemented the rpk mutation were identified. The nucleotide sequence of this region revealed that both Tn5 insertions lay within a gene encoding a protein homologous to the GalR/LacI family of transcriptional regulators that are involved in metabolism. PMID- 9409819 TI - RDH54, a RAD54 homologue in Saccharomyces cerevisiae, is required for mitotic diploid-specific recombination and repair and for meiosis. AB - Most mitotic recombination and repair genes of Saccharomyces cerevisiae show no specificity of action for the genome ploidy. We describe here a novel repair and recombination gene that is specific for recombination and repair between homologous chromosomes. The RDH54 gene is homologous to the RAD54 gene, but rdh54 mutants do not show sensitivity to methyl methanesulfonate at concentrations that sensitize a rad54 mutant. However, the rdh54 null mutation enhances the methyl methanesulfonate sensitivity of a rad54 mutant and single rdh54 mutants are sensitive to prolonged exposure at high concentrations of methyl methanesulfonate. The RDH54 gene is required for recombination, but only in a diploid. We present evidence showing that the RDH54 gene is required for interhomologue gene conversion but not intrachromosomal gene conversion. The rdh54 mutation confers diploid-specific lethalities and reduced growth in various mutant backgrounds. These phenotypes are due to attempted recombination. The RDH54 gene is also required for meiosis as homozygous mutant diploids show very poor sporulation and reduced spore viability. The role of the RDH54 gene in mitotic repair and in meiosis and the pathway in which it acts are discussed. PMID- 9409820 TI - Characterization of the roles of the Saccharomyces cerevisiae RAD54 gene and a homologue of RAD54, RDH54/TID1, in mitosis and meiosis. AB - The RAD54 gene, which encodes a protein in the SWI2/SNF2 family, plays an important role in recombination and DNA repair in Saccharomyces cerevisiae. The yeast genome project revealed a homologue of RAD54, RDH54/TID1. Properties of the rdh54/tid1 mutant and the rad54 rdh54/tid1 double mutant are shown for mitosis and meiosis. The rad54 mutant is sensitive to the alkylating agent, methyl methanesulfonate (MMS), and is defective in interchromosomal and intrachromosomal gene conversion. The rdh54/tid1 single mutant, on the other hand, does not show any significant deficiency in mitosis. However, the rad54 rdh54/tid1 mutant is more sensitive to MMS and more defective in interchromosomal gene conversion than is the rad54 mutant, but shows the same frequency of intrachromosomal gene conversion as the rad54 mutant. These results suggest that RDH54/TID1 is involved in a minor pathway of mitotic recombination in the absence of R4D54. In meiosis, both single mutants produce viable spores at slightly reduced frequency. However, only the rdh54/tid1 mutant, but not the rad54 mutant, shows significant defects in recombination: retardation of the repair of meiosis-specific double-strand breaks (DSBs) and delayed formation of physical recombinants. Furthermore, the rad54 rdh54/tid1 double mutant is completely defective in meiosis, accumulating DSBs with more recessed ends than the wild type and producing fewer physical recombinants than the wild type. These results suggest that one of the differences between the late stages of mitotic recombination and meiotic recombination might be specified by differential dependency on the Rad54 and Rdh54/Tid1 proteins. PMID- 9409822 TI - Control of amino acid permease sorting in the late secretory pathway of Saccharomyces cerevisiae by SEC13, LST4, LST7 and LST8. AB - The SEC13 gene was originally identified by temperature-sensitive mutations that block all protein transport from the ER to the Golgi. We have found that at a permissive temperature for growth, the sec13-1 mutation selectively blocks transport of the nitrogen-regulated amino acid permease, Gap1p, from the Golgi to the plasma membrane, but does not affect the activity of constitutive permeases such as Hip1p, Can1p, or Lyp1p. Different alleles of SEC13 exhibit different relative effects on protein transport from the ER to the Golgi, or on Gap1p activity, indicating distinct requirements for SEC13 function at two different steps in the secretory pathway. Three new genes, LST4, LST7, and LST8, were identified that are also required for amino acid permease transport from the Golgi to the cell surface. Mutations in LST4 and LST7 reduce the activity of the nitrogen-regulated permeases Gap1p and Put4p, whereas mutations in LST8 impair the activities of a broader set of amino acid permeases. The LST8 gene encodes a protein composed of WD-repeats and has a close human homologue. The LST7 gene encodes a novel protein. Together, these data indicate that SEC13, LST4, LST7, and LST8 function in the regulated delivery of Gap1p to the cell surface, perhaps as components of a post-Golgi secretory-vesicle coat. PMID- 9409821 TI - The Saccharomyces cerevisiae RAD30 gene, a homologue of Escherichia coli dinB and umuC, is DNA damage inducible and functions in a novel error-free postreplication repair mechanism. AB - Damage-inducible mutagenesis in prokaryotes is largely dependent upon the activity of the UmuD'C-like proteins. Since many DNA repair processes are structurally and/or functionally conserved between prokaryotes and eukaryotes, we investigated the role of RAD30, a previously uncharacterized Saccharomyces cerevisiae DNA repair gene related to the Escherichia coli dinB, umuC and S. cerevisiae REV1 genes, in UV resistance and UV-induced mutagenesis. Similar to its prokaryotic homologues, RAD30 was found to be damage inducible. Like many S. cerevisiae genes involved in error-prone DNA repair, epistasis analysis clearly places RAD30 in the RAD6 group and rad30 mutants display moderate UV sensitivity reminiscent of rev mutants. However, unlike rev mutants, no defect in UV-induced reversion was seen in rad30 strains. While rad6 and rad18 are both epistatic to rad30, no epistasis was observed with rev1, rev3, rev7 or rad5, all of which are members of the RAD6 epistasis group. These findings suggest that RD30 participates in a novel error-free repair pathway dependent on RAD6 and RAD18, but independent of REV1, REV3, REV7 and RAD5. PMID- 9409824 TI - Mutations synthetically lethal with tpm1delta lie in genes involved in morphogenesis. AB - Yeast contains two genes, TPM1 and TPM2, encoding tropomyosins, either of which can provide an essential function in the yeast cytoskeleton. To elucidate more clearly the function of the major tropomyosin, encoded by TPM1, we have isolated mutations that confer synthetic lethality with the null mutant of TPM1. Here we describe a phenotypic and genetic analysis of mutations in TSL1/BEM2, TSL2, TSL3, TSL5, and TSL6 (tropomyosin synthetic lethal). All the mutants exhibit clear morphological and some actin cytoskeletal defects, but are not noticeably defective in secretion, endocytosis, or organelle segregation. The lethality conferred by tsl tpm1delta mutations could be specifically suppressed by either TPM1 or an additional copy of TPM2. This implies that the essential function compromised in the tsl tpm1delta constructs is the same essential function for which Tpm1p or Tpm2p is necessary. Synthetic interactions and unlinked noncomplementation were observed between the tsl mutants, suggesting that they participate in related functions involving morphogenesis. In support of this, tsl6-1 was identified as an allele of the nonessential gene SLT2 or MPK1 whose product is a MAP kinase regulating cell wall synthesis. These results indicate that this synthetic lethality approach provides a sensitive screen for the isolation of mutations affecting morphogenesis, many of which are likely to be in nonessential genes, like BEM2 and SLT2. PMID- 9409823 TI - The yeast HRS1 gene is involved in positive and negative regulation of transcription and shows genetic characteristics similar to SIN4 and GAL11. AB - We provide genetic evidence that HRS1/PGD1, a yeast gene previously identified as a suppressor of the hyper-recombination phenotype of hpr1, has positive and negative roles in transcriptional regulation. We have analyzed three differently regulated promoters, GAL1, PHO5 and HSP26, by beta-galactosidase assays of lacZ fused promoters and by Northern analysis of the endogenous genes. Transcription of these promoters was derepressed in hrs1delta mutants under conditions in which it is normally repressed in wild type. Under induced conditions it was either strongly reduced or significantly enhanced depending on the promoter system analyzed. Constitutive transcription was not affected, as determined in ADH1 and TEF2. In addition, Hrs1p was required for mating-factor expression, telomere linked DNA silencing and DNA supercoiling of plasmids. Furthermore, hrs1delta suppressed Ty-insertion mutations and conferred a Gal- phenotype. Many of these phenotypes also result from mutations in GAL11, SIN4 or RGR1, which encode proteins of the RNA polII mediator. We also show that gal11delta and sin4delta partially suppress the hyper-rec phenotype of hpr1 mutants, although to a lesser extent than hrs1delta. Our results provide new evidence for the connection between hpr1delta-induced deletions and transcription. We discuss the possibility that Hrs1p might be a component of the RNA polII transcription machinery. PMID- 9409826 TI - Allele-specific suppression by formation of new protein-protein interactions in yeast. AB - Yeast fimbrin is encoded by the SAC6 gene, mutations of which suppress temperature-sensitive mutations in the actin gene (ACT1). To examine the mechanism of suppression, we have conducted a biochemical analysis of the interaction between various combinations of wild-type and mutant actin and Sac6 proteins. Previously, we showed that actin mutations that are suppressed by sac6 mutations encode proteins with a reduced affinity for wild-type Sac6p. In the present study, we have found that mutant Sac6 proteins bind more tightly to mutant actin than does wild-type Sac6p, and thus compensate for weakened interactions caused by the mutant actin. Remarkably, we have also found that mutant Sac6 proteins bind more tightly to wild-type actin than does wild-type Sac6p. This result indicates that suppression does not occur through the restoration of the original contact site, but rather through the formation of a novel contact site. This finding argues against suppression occurring through a "lock-and-key" mechanism and suggests a mechanism involving more global increases in affinity between the two proteins. We propose that the most common kind of suppressors involving interacting proteins will likely occur through this less specific mechanism. PMID- 9409825 TI - Analysis of mutationally altered forms of the Cct6 subunit of the chaperonin from Saccharomyces cerevisiae. AB - The Cct double-ring chaperonin complex of Saccharomyces cerevisiae is comprised of eight essential subunits, Cct1p-Cct8p, and assists the folding of substrates such as actins and tubulins. Single and multiple amino acid replacements of Cct6p were constructed by oligonucleotide-directed mutagenesis, including changes of charged to alanine residues and uncharged to charged residues. The replacements were targeted, in part, to residues corresponding to functionally critical regions identified in the published crystal structure of the Escherichia coli chaperonin, GroEL. Here, we report the critical hydrophobic residues and clusters of hydrophilic residues in regions corresponding to those from the apical domain of GroEL implicated in peptide binding and peptide release, and certain residues in the putative equatorial domain implicated in subunit-to-subunit interaction. In contrast to their homologous counterparts in Cct2p and Cct1p, the highly conserved putative ATP binding motifs of Cct6p were relatively amenable to mutations. Our data suggest that the entire Cct6p molecule might be essential for assembly of Cct complex and might participate in binding substrates. However, there appeared to exist a functional hierarchy in ATP binding/hydrolysis among Cct subunits, as suggested by the high tolerance of Cct6p to mutations within the putative ATP binding pocket. PMID- 9409827 TI - A MADS-box homologue in Ustilago maydis regulates the expression of pheromone inducible genes but is nonessential. AB - Mating and pathogenic development in the smut fungus Ustilago maydis are controlled by a pheromone/receptor system and two homeodomain proteins, bEp and bWp, which form heterodimers in nonallelic combinations. We describe the isolation of a gene, umc1, encoding a MADS-box protein, which displays significant similarity to the Saccharomyces cerevisiae MCM1 gene. umc1 complemented the viability defect of yeast mcm1 mutants. In U. maydis, umc1 deletion mutants were viable and pathogenic development was unaffected. Nevertheless, the basal expression levels of several pheromone-inducible genes were significantly reduced leading to an attenuated mating reaction. In contrast to S. cerevisiae, where Mcm1p plays a crucial role in the cell-type specific expression of a- and alpha-specific genes, the U. maydis umc1 gene appears to have only a modulatory effect on the expression of mating type-specific genes. PMID- 9409829 TI - A limited number of Caenorhabditis elegans genes are readily mutable to dominant, temperature-sensitive maternal-effect embryonic lethality. AB - Dominant gain-of-function mutations can give unique insights into the study of gene function. In addition, gain-of-function mutations, unlike loss-of-function alleles, are not biased against the identification of genetically redundant loci. To identify novel genetic functions active during Caenorhabditis elegans embryogenesis, we have collected a set of dominant temperature-sensitive maternal effect embryonic lethal mutations. In a previous screen, we isolated eight such mutations, distributed among six genes. In the present study, we describe eight new dominant mutations that identify only three additional genes, yielding a total of 16 dominant mutations found in nine genes. Therefore, it appears that a limited number of C. elegans genes mutate to this phenotype at appreciable frequencies. Five of the genes that we identified by dominant mutations have loss of-function alleles. Two of these genes may lack loss-of-function phenotypes, indicating that they are nonessential and so may represent redundant loci. Loss of-function mutations of three other genes are associated with recessive lethality, indicating nonredundancy. PMID- 9409828 TI - Genetic analysis of the Chlamydomonas reinhardtii I-CreI mobile intron homing system in Escherichia coli. AB - We have developed and used a genetic selection system in Escherichia coli to study functional requirements for homing site recognition and cleavage by a representative eukaryotic mobile intron endonuclease. The homing endonuclease, I CreI, was originally isolated from the chloroplast of the unicellular green alga Chlamydomonas reinhardtii. I-CreI homing site mutants contained base pair substitutions or single base deletions that altered the rate of homing site cleavage and/or product release. I-CreI endonuclease mutants fell into six phenotypic classes that differed in in vivo activity, toxicity or genetic dominance. Inactivating mutations clustered in the N-terminal 60% of the I-CreI amino acid sequence, and two frameshift mutations were isolated that resulted in premature translation termination though retained partial activity. These mutations indicate that the N-terminal two-thirds of the I-CreI endonuclease is sufficient for homing site recognition and cleavage. Substitution mutations altered in four potential active site residues were examined: D20N, Q47H or R70A substitutions inactivated endonuclease activity, whereas S22A did not. The genetic approach we have taken complements phylogenetic and structural studies of mobile intron endonucleases and has provided new information on the mechanistic basis of I-CreI homing site recognition and cleavage. PMID- 9409830 TI - Identification and characterization of genes that interact with lin-12 in Caenorhabditis elegans. AB - We identified and characterized 14 extragenic mutations that suppressed the dominant egg-laying defect of certain lin-12 gain-of-function mutations. These suppressors defined seven genes: sup-17, lag-2, sel-4, sel-5, sel-6, sel-7 and sel-8. Mutations in six of the genes are recessive suppressors, whereas the two mutations that define the seventh gene, lag-2, are semi-dominant suppressors. These suppressor mutations were able to suppress other lin-12 gain-of-function mutations. The suppressor mutations arose at a very low frequency per gene, 10-50 times below the typical loss-of-function mutation frequency. The suppressor mutations in sup-17 and lag-2 were shown to be rare non-null alleles, and we present evidence that null mutations in these two genes cause lethality. Temperature-shift studies for two suppressor genes, sup-17 and lag-2, suggest that both genes act at approximately the same time as lin-12 in specifying a cell fate. Suppressor alleles of six of these genes enhanced a temperature-sensitive loss-of-function allele of glp-1, a gene related to lin-12 in structure and function. Our analysis of these suppressors suggests that the majority of these genes are part of a shared lin-12/glp-1 signal transduction pathway, or act to regulate the expression or stability of lin-12 and glp-1. PMID- 9409831 TI - P-element insertion alleles of essential genes on the third chromosome of Drosophila melanogaster: correlation of physical and cytogenetic maps in chromosomal region 86E-87F. AB - We have established a collection of 2460 lethal or semi-lethal mutant lines using a procedure thought to insert single P elements into vital genes on the third chromosome of Drosophila melanogaster. More than 1200 randomly selected lines were examined by in situ hybridization and 90% found to contain single insertions at sites that mark 89% of all lettered subdivisions of the Bridges' map. A set of chromosomal deficiencies that collectively uncover approximately 25% of the euchromatin of chromosome 3 reveal lethal mutations in 468 lines corresponding to 145 complementation groups. We undertook a detailed analysis of the cytogenetic interval 86E-87F and identified 87 P-element-induced mutations falling into 38 complementation groups, 16 of which correspond to previously known genes. Twenty one of these 38 complementation groups have at least one allele that has a P element insertion at a position consistent with the cytogenetics of the locus. We have rescued P elements and flanking chromosomal sequences from the 86E-87F region in 35 lines with either lethal or genetically silent P insertions, and used these as probes to identify cosmids and P1 clones from the Drosophila genome projects. This has tied together the physical and genetic maps and has linked 44 previously identified cosmid contigs into seven "super-contigs" that span the interval. STS data for sequences flanking one side of the P-element insertions in 49 lines has identified insertions in the alphagamma element at 87C, two known transposable elements, and the open reading frames of seven putative single copy genes. These correspond to five known genes in this interval, and two genes identified by the homology of their predicted products to known proteins from other organisms. PMID- 9409832 TI - P-element insertion alleles of essential genes on the third chromosome of Drosophila melanogaster: mutations affecting embryonic PNS development. AB - To identify novel genes and to isolate tagged mutations in known genes that are required for the development of the peripheral nervous system (PNS), we have screened a novel collection of 2460 strains carrying lethal or semilethal P element insertions on the third chromosome. Monoclonal antibody 22C10 was used as a marker to visualize the embryonic PNS. We identified 109 mutant strains that exhibited reproducible phenotypes in the PNS. Cytological and genetic analyses of these strains indicated that 87 mutations affect previously identified genes: tramtrack (n = 18 alleles), string (n = 15), cyclin A (n = 13), single-minded (n = 13), Delta (n = 9), neuralized (n = 4), pointed (n = 4), extra macrochaetae (n = 4), prospero (n = 3), tartan (n = 2), and pebble (n = 2). In addition, 13 mutations affect genes that we identified recently in a chemical mutagenesis screen designed to isolate similar mutants: hearty (n = 3), dorsotonals (n = 2), pavarotti (n = 2), sanpodo (n = 2), dalmatian (n = 1), missensed (n = 1), senseless (n = 1), and sticky ch1 (n = 1). The remaining nine mutations define seven novel complementation groups. The data presented here demonstrate that this collection of P elements will be useful for the identification and cloning of novel genes on the third chromosome, since >70% of mutations identified in the screen are caused by the insertion of a P element. A comparison between this screen and a chemical mutagenesis screen undertaken earlier highlights the complementarity of the two types of genetic screens. PMID- 9409833 TI - Drosophila male-specific lethal-2 protein: structure/function analysis and dependence on MSL-1 for chromosome association. AB - MSL-2 is required for the male-specific assembly of a dosage compensation regulatory complex on the X chromosome of Drosophila melanogaster. We found that MSL-2 binds in a reproducible, partial pattern to the male X chromosome in the absence of MLE or MSL-3, or when ectopically expressed at a low level in females. Moreover, the pattern of MSL-2 binding corresponds precisely in each case to that of MSL-1, suggesting that the two proteins function together to associate with the X. Consistent with this hypothesis, we isolated EMS-induced loss of function msl-1 and msl-2 alleles in a screen for suppressors of the toxic effects of MSL-2 expression in females. We also used site-directed mutagenesis to determine the importance of the MSL-2 RING finger domain and second cysteine-rich motif. The mutations, including those in conserved zinc coordinating cysteines, confirm that the RING finger is essential for MSL-2 function, while suggesting a less stringent requirement for an intact second motif. PMID- 9409834 TI - The congested-like tracheae gene of Drosophila melanogaster encodes a member of the mitochondrial carrier family required for gas-filling of the tracheal system and expansion of the wings after eclosion. AB - A recessive semi-lethal mutation resulting from the insertion of a P-lacW transposon at the cytological position 23A on the polytene chromosomes of Drosophila melanogaster was found to affect the unfolding and expansion of the wings resulting in a loss of venation and a marked decrease in their size. Lethality was polyphasic with numerous animals dying during early larval development and displaying apparently collapsed tracheal trees. The gene was therefore designated as congested-like tracheae, or colt. The colt mutation resulted from the insertion of a P-lacW transposon within the coding region of a 1.4-kb transcript. Wild-type function was restored by inducing a precise excision of the P-lacW transposon, while a deletion of the colt locus, produced by imprecise excision of the P element, showed a phenotype similar to that of the original P insert. The colt gene consists of a single exon and encodes a protein of 306 amino acids made of three tandem repeats, each characterized by two predicted transmembrane segments and a loop domain. The COLT protein shares extensive homology with proteins in the mitochondrial carrier family and particularly with the DIF-1 protein of Caenorhabditis elegans, which has been shown to be maternally required for embryonic tissue differentiation. Our analysis revealed that zygotic colt function is dispensable for normal embryonic morphogenesis but is required for gas-filling of the tracheal system at hatching time of the embryo and for normal epithelial morphogenesis of the wings. PMID- 9409835 TI - The accumulation of P-element-induced recombinants in the germline of male Drosophila melanogaster. AB - P-element-induced recombination in Drosophila melanogaster occurs premeiotically. Recombinants are therefore expected to accumulate in the stem cells of the germline of P-element-carrying males. We show that both the recombination frequency and the incidence of "clustering" increase with the age of males carrying various P-element derivatives. The combination of end-deleted elements can lead to average recombination frequencies >50% with individual instances of 100% recombination. These elements also lowered the fertility of the carriers. We investigated these features by constructing an analytical and a computer simulation model of the course of events in the germline, incorporating the recently proposed hybrid element insertion (HEI) model of P-element activity. The model is able to predict extreme recombination levels, segregation ratio biases and lowered fertility through cell death in a single analysis. PMID- 9409836 TI - Calmodulin point mutations affect Drosophila development and behavior. AB - Calmodulin (CAM) is recognized as a major intermediary in intracellular calcium signaling, but as yet little is known of its role in developmental and behavioral processes. We have generated and studied mutations to the endogenous Cam gene of Drosophila melanogaster that change single amino acids within the protein coding region. One of these mutations produces a striking pupal lethal phenotype involving failure of head eversion. Various mutant combinations produce specific patterns of ectopic wing vein formation or melanotic scabs on the cuticle. Anaphase chromosome bridging is also seen as a maternal effect during the early embryonic nuclear divisions. In addition, specific behavioral defects such as poor climbing and flightlessness are detected among these mutants. Comparisons with other Drosophila mutant phenotypes suggests potential CAM targets that may mediate these developmental and behavioral effects, and analysis of the CAM crystal structure suggests the structural consequences of the individual mutations. PMID- 9409837 TI - Haldane's rule and X-chromosome size in Drosophila. AB - The "dominance theory" of HALDANE'S rule postulates that hybrids of the heterogametic sex are more likely to be inviable or sterile than the homogametic sex because some of the epistatic incompatibilities contributing to postzygotic isolation behave as X-linked partial recessives. When this is true, pairs of taxa with relatively large X chromosomes should require less divergence time, on average, to produce HALDANE'S rule than pairs with smaller Xs. Similarly, if the dominance theory is correct and if the X chromosome evolves at a similar rate to the autosomes, the size of the X should not influence the rate at which homogametic hybrids become inviable or sterile. We use Drosophila data to examine both of these predictions. As expected under the dominance theory, pairs of taxa with large X chromosomes (approximately 40% of the nuclear genome) show HALDANE's rule for sterility at significantly smaller genetic distances than pairs with smaller X chromosomes (approximately 20% of the genome). As also predicted, the genetic distances between taxa that exhibit female inviability/sterility show no differences between "large X" vs. "small X" pairs. We present some simple mathematical models to relate these data to the dominance theory and alternative hypotheses involving faster evolution of the X vs. the autosomes and/or faster evolution of incompatibilities that produce male-specific vs. female-specific sterility. Although the data agree qualitatively with the predictions of the dominance theory, they depart significantly from the quantitative predictions of simple models of the dominance theory and the other hypotheses considered. These departures probably stem from the many simplifying assumptions needed to tractably model epistatic incompatibilities and to analyze heterogeneous data from many taxa. PMID- 9409838 TI - Patterns of mitochondrial variation within and between African malaria vectors, Anopheles gambiae and An. arabiensis, suggest extensive gene flow. AB - Anopheles gambiae and An. arabiensis are mosquito species responsible for most malaria transmission in sub-Saharan Africa. They are also closely related sibling species that share chromosomal and molecular polymorphisms as a consequence of incomplete lineage sorting or introgressive hybridization. To help resolve these processes, this study examined the partitioning of mtDNA sequence variation within and between species across Africa, from both population genetic and phylogeographic perspectives. Based on partial gene sequences from the cytochrome b, ND1 and ND5 genes, haplotype diversity was high but sequences were very closely related. Within species, little or no population subdivision was detected, and there was no evidence for isolation by distance. Between species, there were no fixed nucleotide differences, a high proportion of shared polymorphisms, and eight haplotypes in common over distances as great as 6000 km. Only one of 16 shared polymorphisms led to an amino acid difference, and there was no compelling evidence for nonneutral variation. Parsimony networks constructed of haplotypes from both species revealed no correspondence of haplotype with either geography or taxonomy. This trend of low intraspecific genetic divergence is consistent with evidence from allozyme and microsatellite data and is interpreted in terms of both extensive gene flow and recent range expansion from relatively large, stable populations. We argue that retention of ancestral polymorphisms is a plausible but insufficient explanation for low interspecific genetic divergence, and that extensive hybridization is a contributing factor. PMID- 9409841 TI - The sampling distribution of disease-associated alleles. AB - A theory is developed that provides the sampling distribution of low frequency alleles at a single locus under the assumption that each allele is the result of a unique mutation. The numbers of copies of each allele is assumed to follow a linear birth-death process with sampling. If the population is of constant size, standard results from theory of birth-death processes show that the distribution of numbers of copies of each allele is logarithmic and that the joint distribution of numbers of copies of k alleles found in a sample of size n follows the Ewens sampling distribution. If the population from which the sample was obtained was increasing in size, if there are different selective classes of alleles, or if there are differences in penetrance among alleles, the Ewens distribution no longer applies. Likelihood functions for a given set of observations are obtained under different alternative hypotheses. These results are applied to published data from the BRCA1 locus (associated with early onset breast cancer) and the factor VIII locus (associated with hemophilia A) in humans. In both cases, the sampling distribution of alleles allows rejection of the null hypothesis, but relatively small deviations from the null model can account for the data. In particular, roughly the same population growth rate appears consistent with both data sets. PMID- 9409839 TI - The autosomal chorion locus of the medfly Ceratitis capitata. I. Conserved synteny, amplification and tissue specificity but sequence divergence and altered temporal regulation. AB - We report the isolation, full sequence characterization, amplification and expression properties of medfly chorion genes corresponding to the autosomal chorion locus of Drosophila. These genes are found adjacent to the paramyosin gene and are organized in the same order and tandem orientation as their Drosophila homologues, although they are spaced further apart. They show substantial sequence divergence from their Drosophila homologues, including novel peptide repeats and a new spacing of the tyrosines, which are known to be cross linked in Dipteran chorion. The genes are amplified and expressed during oogenesis, as in Drosophila. Three of them are expressed in the same relative temporal order as in Drosophila but the fourth gene, the homologue of s15, shows a clear shift to an earlier expression period. This is the first known instance of changed temporal regulation in dipteran chorion genes. PMID- 9409840 TI - Geographic structure of mitochondrial and nuclear gene polymorphisms in Australian green turtle populations and male-biased gene flow. AB - The genetic structure of green turtle (Chelonia mydas) rookeries located around the Australian coast was assessed by (1) comparing the structure found within and among geographic regions, (2) comparing microsatellite loci vs. restriction fragment length polymorphism analyses of anonymous single copy nuclear DNA (ascnDNA) loci, and (3) comparing the structure found at nuclear DNA markers to that of previously analyzed mitochondrial (mtDNA) control region sequences. Significant genetic structure was observed over all regions at both sets of nuclear markers, though the microsatellite data provided greater resolution in identifying significant genetic differences in pairwise tests between regions. Inferences about population structure and migration rates from the microsatellite data varied depending on whether statistics were based on the stepwise mutation or infinite allele model, with the latter being more congruent with geography. Estimated rates of gene flow were generally higher than expected for nuclear DNA (nDNA) in comparison to mtDNA, and this difference was most pronounced in comparisons between the northern and southern Great Barrier Reef (GBR). The genetic data combined with results from physical tagging studies indicate that the lack of nuclear gene divergence through the GBR is likely due to the migration of sGBR turtles through the courtship area of the nGBR population, rather than male-biased dispersal. This example highlights the value of combining comparative studies of molecular variation with ecological data to infer population processes. PMID- 9409842 TI - Nucleotide sequence evolution at the kappa-casein locus: evidence for positive selection within the family Bovidae. AB - Kappa-casein is a mammalian milk protein involved in a number of important physiological processes. In the gut, the ingested protein is split into an insoluble peptide (para kappa-casein) and a soluble hydrophilic glycopeptide (caseinomacropeptide). Caseinomacropeptide is responsible for increased efficiency of digestion, prevention of neonate hypersensitivity to ingested proteins, and inhibition of gastric pathogens. Variation within this peptide has significant effects associated with important traits such as milk production. The nucleotide sequences for regions of kappa-casein exon and intron four were determined for representatives of the artiodactyl family Bovidae. The pattern of nucleotide substitution in kappa-casein sequences for distantly related bovid taxa demonstrates that positive selection has accelerated their divergence at the amino acid sequence level. This selection has differentially influenced the molecular evolution of the two kappa-casein split peptides and is focused within a 34-codon region of caseinomacropeptide. PMID- 9409843 TI - Evolutionary divergence of the genetic architecture underlying photoperiodism in the pitcher-plant mosquito, Wyeomyia smithii. AB - We determine the contribution of composite additive, dominance, and epistatic effects to the genetic divergence of photoperiodic response along latitudinal, altitudinal, and longitudinal gradients in the pitcher-plant mosquito, Wyeomyia smithii. Joint scaling tests of crosses between populations showed widespread epistasis as well as additive and dominance differences among populations. There were differences due to epistasis between an alpine population in North Carolina and populations in Florida, lowland North Carolina, and Maine. Longitudinal displacement resulted in differences due to epistasis between Florida and Alabama populations separated by 300 km but not between Maine and Wisconsin populations separated by 2000 km. Genetic differences between New Jersey and Ontario did not involve either dominance or epistasis and we estimated the minimum number of effective factors contributing to a difference in mean critical photoperiod of 5 SD between them as nE = 5. We propose that the genetic similarity of populations within a broad northern region is due to their more recent origin since recession of the Laurentide Ice Sheet and that the unique genetic architecture of each population is the result of both mutation and repeated migration-founder-flush episodes during the dispersal of W. smithii in North America. Our results suggest that differences in composite additive and dominance effects arise early in the genetic divergence of populations while differences due to epistasis accumulate after more prolonged isolation. PMID- 9409844 TI - The TRANSFORMER genes of the fern Ceratopteris simultaneously promote meristem and archegonia development and repress antheridia development in the developing gametophyte. AB - The sex of the haploid gametophyte of the fern Ceratopteris is determined by the presence or absence of the pheromone antheridiogen, which, when present, promotes male development and represses female development of the gametophyte. Several genes involved in sex determination in Ceratopteris have been identified by mutation. In this study, the epistatic interactions among new and previously described sex-determining mutants have been characterized. These results show that sex expression is regulated by two sets of genes defined by the FEM1 and TRA loci. Each promotes the expression of either male or female traits and simultaneosly represses the expression of the other. A model describing how antheridiogen regulates the expression of these genes and the sex of the gametophyte is described. The observation that some gametophytic sex-determining mutants have phenotypic effects on the sporophyte plant indicates that sex determination in the Ceratopteris gametophyte is regulated by a mechanism that also regulates sporophyte development. PMID- 9409845 TI - DNA polymorphism at the Pgi locus of a wild yam, Dioscorea tokoro. AB - To study the origin and maintenance mechanisms of the PGI allozyme polymorphism of a wild plant, Dioscorea tokoro, DNA sequences of the entire coding region (1701 bp) and two intronic regions (total 2049 bp) of the Pgi gene as well as a part of the Adh gene (590 bp) were analyzed. Two replacement substitutions were revealed to be responsible for the differentiation of three allozymes alleles (Pgi-a, Pgi-b and Pgi-c) that occur in natural population in intermediate frequencies. Interspecific comparison of DNA sequences identified Pgi-b as the oldest allele, from which two other alleles were derived probably within the last 150,000 years. The level of DNA polymorphism at D. tokoro Pgi locus was low. No elevated level of DNA polymorphism was detected in the close vicinity of the two replacement sites differentiating the three allozymes. Departures from the neutral mutation hypothesis were detected by Fu and Li's and MK tests. The observed patterns of DNA polymorphism are explainable by both (1) the neutral mutation hypothesis with an assumption of small effective size of D. tokoro population, and (2) the positive selection hypothesis that the allele frequencies of Pgi-a and Pgi-c have increased in a short time by their selective advantages. PMID- 9409846 TI - B chromosome behavior in maize pollen as determined by a molecular probe. AB - The B chromosomes of maize typically undergo nondisjunction during the second microspore division (generative cell division). When the microspore nucleus contains only one B chromosome, two kinds of sperm result, one with two B chromosomes and one with no B chromosomes. The sperm with the B chromosomes preferentially fertilizes the egg cell. Previous studies of these phenomena have been limited to genetic analysis and chromosome spreads. In this study we show that a B chromosome-specific probe can be used with fluorescence in situ hybridization (FISH) analysis to detect the presence, location, and frequency of B chromosomes in intact interphase nuclei within mature pollen of maize. Using genetic line TB-10L18, our results indicate that nondisjunction of the B centromere occurs at an average frequency of 56.6%, based on four plants and 1306 pollen grains analyzed. This is consistent with the results of genetic studies using the same B-A translocation. In addition, our results suggest that B chromosome nondisjunction can occur during the first microspore division. Spatial distribution of the B chromosome-specific probe appears to be largely confined to one tip of the sperm nucleus, and a DNA fragment found outside the pollen nuclei often hybridizes to the B chromosome-specific probe. PMID- 9409847 TI - Germinal excisions of the maize transposon activator do not stimulate meiotic recombination or homology-dependent repair at the bz locus. AB - Double-strand breaks have been implicated both in the initiation of meiotic recombination in yeast and as intermediates in the transposition process of nonreplicative transposons. Some transposons of this class, notably P of Drosophila and Tc1 of Caenorhabditis elegans, promote a form of homology dependent premeiotic gene conversion upon excision. In this work, we have looked for evidence of an interaction between Ac transposition and meiotic recombination at the bz locus in maize. We find that the frequency of meiotic recombination between homologues is not enhanced by the presence of Ac in one of the bz heteroalleles and, conversely, that the presence of a homologous sequence in either trans (homologous chromosome) or cis (tandem duplication) does not promote conversion of the Ac insertion site. However, a tandem duplication of the bz locus may be destabilized by the insertion of Ac. We discuss possible reasons for the lack of interaction between Ac excision and homologous meiotic recombination in maize. PMID- 9409848 TI - Bayesian procedures for discriminating among hypotheses with discrete distributions: inheritance in the tetraploid Astilbe biternata. AB - Discrimination between disomic and tetrasomic inheritance aids in determining whether tetraploids originated by allotetraploidy or autotetraploidy, respectively. Past assessments of inheritance in tetraploids have used analyses whereby each inheritance hypothesis is tested independently. I present a Bayesian analysis that is appropriate for discriminating among several inheritance hypotheses and can be used in any case where hypotheses are defined by discrete distributions. The Bayesian approach incorporates prior knowledge of the probability of occurrence of disomic and tetrasomic hypotheses so that the results of the analysis are not biased by the fact that there is a single tetrasomic hypothesis and multiple disomic hypotheses. This analysis is used to interpret data from crosses in the tetraploid Astilbe biternata, a herbaceous plant native to the southern Appalachians. The progeny ratios from all crosses favored the hypothesis of disomic inheritance at both the PGM and slow-PGI loci. These results support earlier cytogenetic evidence for the allotetraploid origin of Astilbe biternata. PMID- 9409850 TI - Estimation of the amount of DNA polymorphism when the neutral mutation rate varies among sites. AB - Knowing the amount of DNA polymorphism is essential to understand the mechanism of maintaining DNA polymorphism in a natural population. The amount of DNA polymorphism can be measured by the average number of nucleotide differences per site (pi), the proportion of segregating (polymorphic) site (s) and the minimum number of mutations per site (s*). Since the latter two quantities depend on the sample size, theta is often used as a measure of the amount of DNA polymorphism, where theta = 4Nmu, N is the effective population size and mu is the neutral mutation rate per site per generation. It is known that theta estimated from pi, s and s* under the infinite site model can be biased when the mutation rate varies among sites. We have therefore developed new methods for estimating theta under the finite site model. Using computer simulations, it has been shown that the new methods give almost unbiased estimates even when the mutation rate varies among sites substantially. Furthermore, we have also developed new statistics for testing neutrality by modifying Tajima's D statistic. Computer simulations suggest that the new test statistics can be used even when the mutation rate varies among sites. PMID- 9409849 TI - An empirical evaluation of genetic distance statistics using microsatellite data from bear (Ursidae) populations. AB - A large microsatellite data set from three species of bear (Ursidae) was used to empirically test the performance of six genetic distance measures in resolving relationships at a variety of scales ranging from adjacent areas in a continuous distribution to species that diverged several million years ago. At the finest scale, while some distance measures performed extremely well, statistics developed specifically to accommodate the mutational processes of microsatellites performed relatively poorly, presumably because of the relatively higher variance of these statistics. At the other extreme, no statistic was able to resolve the close sister relationship of polar bears and brown bears from more distantly related pairs of species. This failure is most likely due to constraints on allele distributions at microsatellite loci. At intermediate scales, both within continuous distributions and in comparisons to insular populations of late Pleistocene origin, it was not possible to define the point where linearity was lost for each of the statistics, except that it is clearly lost after relatively short periods of independent evolution. All of the statistics were affected by the amount of genetic diversity within the populations being compared, significantly complicating the interpretation of genetic distance data. PMID- 9409851 TI - Hardy-Weinberg testing for continuous data. AB - Estimation of allelic and genotypic distributions for continuous data using kernel density estimation is discussed and illustrated for some variable number of tandem repeat data. These kernel density estimates provide a useful representation of data when only some of the many variants at a locus are present in a sample. Two Hardy-Weinberg test procedures are introduced for continuous data: a continuous chi-square test with test statistic T(CCS) and a test based on Hellinger's distance with test statistic T(HD). Simulations are used to compare the powers of these tests to each other and to the powers of a test of intraclass correlation T(IC), as well as to the power of Fisher's exact test T(FET) applied to discretized data. Results indicate that the power of T(CCS) is better than that of T(HD), but neither is as powerful as T(FET). The intraclass correlation test does not perform as well as the other tests examined in this article. PMID- 9409852 TI - Alu evolution in human populations: using the coalescent to estimate effective population size. AB - There are estimated to be approximately 1000 members of the Ya5 Alu subfamily of retroposons in humans. This subfamily has a distribution restricted to humans, with a few copies in gorillas and chimpanzees. Fifty-seven Ya5 elements were previously cloned from a HeLa-derived randomly sheared total genomic library, sequenced, and screened for polymorphism in a panel of 120 unrelated humans. Forty-four of the 57 cloned Alu repeats were monomorphic in the sample and 13 Alu repeats were dimorphic for insertion presence/absence. The observed distribution of sample frequencies of the 13 dimorphic elements is consistent with the theoretical expectation for elements ascertained in a single diploid cell line. Coalescence theory is used to compute expected total pedigree branch lengths for monomorphic and dimorphic elements, leading to an estimate of human effective population size of approximately 18,000 during the last one to two million years. PMID- 9409853 TI - Differentiating between selection and mutation bias. PMID- 9409854 TI - Distinguishing the effects of mutational biases and natural selection on DNA sequence variation. PMID- 9409855 TI - Transposable element distributions in Drosophila. PMID- 9409856 TI - Transposable element distribution in Drosophila. PMID- 9409857 TI - Differential increases in AFP, hCG, and uE3 in twin pregnancies: impact on attempts to quantify Down syndrome screening calculations. AB - Since the advent of multiple marker screening (MMS) for Down syndrome (DS) risk calculations, limitations for twins have been apparent. Recent attempts have been made to extrapolate mathematically singleton risks to twins. Here we investigate the pattern of levels among AFP, hCG, and uE3 in twins. MMS screening data from 4,443 twin pregnancies were compared to those from 258,885 singletons from 14-21 weeks of gestational age during a 3-year period (1992-1994) in our laboratory. Medians were determined for singletons and twins, and the ratios of twins to singletons were derived. Median AFP levels for twins are approximately double those of singletons, but median increases for hCG and uE3 are less than double. The data were divided further by ethnic groups (white, African American, Asian, and Hispanic), among which there were significant variations in medians, but not in the ratios of twins to singletons. The increased serum levels of different markers in twins are not consistent across analytes, possibly reflecting independent development of different compartments. Such differences mean that a mere mathematical conversion of singleton DS risks would be imbalanced among the analytes and cannot be applied reasonably to twins. Ethnic-specific databases are as important in twins as they are in singletons. PMID- 9409858 TI - Elevated rates of severe neural tube defects in a high-prevalence area in northern China. AB - In the northern provinces of China, the birth prevalence rate of neural tube defects (NTDs) is among the highest in the world-at about 6 per 1,000 births in rural areas. A unique population-based birth defects surveillance system in which photographs are taken of infants with selected external birth defects was implemented in two provinces in northern China and two provinces in southern China where NTD rates approximate those in the United States. In the period from March 1992 through December 1993, 660 infants with NTDs were identified by the surveillance project from a birth cohort of 251,567. We compared data from the two surveillance areas in China with data from a low-prevalence area in the United States to determine if the pattern of NTD types differs. Based on birth prevalence rates of NTDs from the Metropolitan Atlanta Congenital Defects Program, the observed to expected ratios for two types of NTDs are markedly increased at 80.8 for craniorachischisis and 25.0 for iniencephaly. Rates of these two NTDs in the southern provinces are increased to a lesser degree with observed to expected ratios of 7.1 for craniorachischisis and 2.7 for iniencephaly. The pattern of NTDs in northern China shows an increase in types that are rare in low-prevalence areas such as metropolitan Atlanta. Increased awareness of varying patterns of NTDs in different populations may have important implications for identifying etiologic and pathogenetic mechanisms of NTDs. PMID- 9409859 TI - Familial tandem duplication of bands q31.1 to q32.3 on chromosome 4 with mild phenotypic effect. AB - We describe a family carrying a direct duplication of an interstitial segment of the distal long arm of chromosome 4(q31.1q32.3) with a mild clinical phenotype. Affected family members include a 27-year-old woman and 2 of her 4 children, 1 of whom also has Klinefelter syndrome. Although 8 cases of simple duplications or insertions of 4q material have been described, only 3 include bands q31-q32, and these involve additional adjacent material. Affected members of the pedigree reported here have some of the manifestations of previously described cases of duplication 4q, but are less severely affected, suggesting that the genetic material in this region is well tolerated as a partial trisomy. PMID- 9409860 TI - Arrhythmogenic right ventricular dysplasia and anterior polar cataract. AB - Arrythmogenic right ventricular dysplasia (ARVD) is an autosomal dominant inherited cardiomyopathy with incomplete penetrance and variable expressivity. Recently, the gene was mapped to 14q23-24. It is being increasingly investigated as a major cause of sudden death at a young age. Anterior polar cataract (APC) is a rare hereditary form of lens opacity. The locus for an APC gene was located tentatively on 14q24qter. We describe a patient with a severe form of ARVD in whom asymptomatic APC was detected by an ophthalmologic examination. His sister had ARVD and similar cataracts. Parents were second cousins but were healthy. This is the first report of possible autosomal recessive inheritance of ARVD. This is also the first time that the combination of ARVD and APC is reported. Three possibilities may explain this concurrence: pleiotropy, contiguous gene syndrome, or coincidence. Our findings suggest placement of an APC gene at 14q23 24. PMID- 9409861 TI - Private multiple congenital anomaly syndromes may result from unbalanced subtle translocations: t(2q;4p) explains the Lambotte syndrome. AB - In 1990, Lambotte syndrome was reported as an apparently autosomal recessive multiple congenital anomaly/mental retardation (MCA/MR) syndrome observed in 4 of 12 sibs from a probably consanguineous mating [Verloes et al., Am J Med Genet 1990; 37:119-123]. Major manifestations included intrauterine growth retardation (IUGR), microcephaly, large soft pinnae, hypertelorism, beaked nose, and extremely severe neurologic impairment, with holoprosencephaly in one instance. After the observation of a further affected child born of one unaffected sister, in situ hybridization analysis and chromosome painting techniques demonstrated a subtle t(2;4)(q37.1; p16.2) translocation in the mother, suggesting a combination of 2q/4p trisomy/monosomy in all of the affected children of the family. Many private sporadic or recurrent MCA/MR syndromes maybe due to similar symmetric translocations, undetectable by conventional banding techniques. PMID- 9409862 TI - Atypical skeletal changes in otopalatodigital syndrome type II: phenotypic overlap among otopalatodigital syndrome type II, boomerang dysplasia, atelosteogenesis type I and type III, and lethal male phenotype of Melnick Needles syndrome. AB - We report on 2 cases of otopalatodigital syndrome type II (OPD II) with atypical skeletal changes, overlapping those of boomerang dysplasia, atelosteogenesis type I (AO I) and type III (AO III), and the lethal male phenotype of Melnick-Needles syndrome. One patient exhibited strikingly broad, bowed femora, which resembled those of boomerang dysplasia. The other patient possessed conspicuous undertubulation of the long bones, defective ossification of the spine, and severe undermineralization of the calvaria, which may have caused diagnostic confusion with AO I, AO III, and the lethal male phenotype of Melnick-Needles syndrome. OPD II is transmitted as an X-linked recessive trait, whereas AO I, AO III, and boomerang dysplasia are considered to result from a new dominant mutation, and Melnick-Needles syndrome is inherited as an X-linked dominant trait. Accordingly, differential diagnosis is mandatory to provide the affected families with adequate genetic counseling. Awareness of these skeletal changes in OPD II will prevent the misdiagnosis of this entity as other disorders. Furthermore, the phenotypic overlap among these disorders may expand the entities that constitute the OPD-Larsen dysplasia family proposed by Spranger [1985]. PMID- 9409863 TI - X-linked recessive chondrodysplasia punctata due to a new point mutation of the ARSE gene. AB - Chondrodysplasia punctata (CP) is a heterogeneous group of bone dysplasias that are characterized by abnormal calcium deposition in areas of enchondral bone formation. The existence of an X-linked recessive form of chondrodysplasia punctata (CDPX) has been recognized in patients who are nullisomic for the Xp22.3 region, presenting with complex phenotypes. The gene of CDPX has been identified recently, and five point mutations of the gene, named ARSE, have been described. Here, we report on the clinical and molecular characterization of a patient with CDPX. The patient presented at birth with cranial and facial anomalies and short stature; an x-ray skeletal survey showed punctate calcifications and striking hand and foot abnormalities. Single strand conformation polymorphism (SSCP) and sequence analysis of the patient's DNA allowed the identification of a new mutation of the ARSE gene; this mutation causes an amino acid substitution from cysteine to tyrosine at position 492 of the ARSE predicted protein product. The clinical description of patients with CDPX due to known mutation of the ARSE is of interest for the precise delineation of the clinical spectrum of the disease. PMID- 9409864 TI - Severe brain and limb defects with possible autosomal recessive inheritance: a series of six cases and review of the literature. AB - Six fetuses with normal chromosomes were found to have severe craniofacial, limb, and visceral malformations during the second trimester of pregnancy. Two of these fetuses were monozygotic twins while a third one had a healthy dizygotic twin brother. A case with familial recurrence was also observed. Autopsy and skeletal radiographs suggested several diagnoses such as neural tube defect with limb defects or XK aprosencephaly. The development of these severe conditions in monozygotic twins and familial recurrence emphasize the difficulties of genetic counseling in such situations. These cases may suggest autosomal recessive inheritance. PMID- 9409865 TI - Genealogy, natural history, and phenotype of Alstrom syndrome in a large Acadian kindred and three additional families. AB - We describe a large Acadian kindred including 8 Alstrom Syndrome (AS) patients, with an age range of 4 to 26 at the time of clinical assessment. The affected subjects come from 5 nuclear families within this kindred. The phenotype includes early childhood retinopathy, progressive sensorineural hearing loss, truncal obesity, and acanthosis nigricans. In addition, hyperinsulinemia and hypertriglyceridemia with normal cholesterol levels were observed in most affected individuals tested. Non-insulin dependent diabetes mellitus and growth retardation appear to be age-related manifestations that occur post-adolescence. Younger affected children are not overtly hyperglycemic and are normal or above average height for age. Although the AS patients in kindred 1 presumably carry the same mutation, many manifestations of the disease are variable. For example, of the 8 children in the Acadian kindred, 4 have scoliosis, 2 have had infantile cardiomyopathy, 2 are hypothyroid, 1 has had hepatic dysfunction and is hypertensive, and 4 have developed asthma. Seven subjects described in this kindred exhibit developmental delay. One additional manifestation not described widely in the literature, advanced bone age, was observed in all subjects tested. The clinical data from this large Acadian kindred, together with information obtained from 4 additional AS patients in 3 unrelated kindreds, confirm and extend clinical observations previously described. In addition, the Acadian kindred with multiple affected individuals, probably arising from a common founder, should allow for identification of the chromosomal localization of a gene causing AS. PMID- 9409866 TI - VATERL: an epidemiologic analysis of risk factors. AB - This work analyzed the incidence of risk factors in 138 cases presenting two or more of five components defining VATERL, with no other recognized unrelated anomalies: vertebral anomalies, anal atresia, esophageal atresia with or without tracheoesophageal fistula, renal anomalies, and preaxial defects of the upper limbs, including polydactyly of the thumb. The 138 infants were ascertained among 1,811,461 births examined in the 1967-1994 period by the Latin-American Collaborative Study of Congenital Malformations: ECLAMC. One healthy and one malformed control newborn infant were matched to each VATERL case. The birth prevalence rates (per 100,000 births) for VATERL were significantly lower in Venezuela (3.1) than in the other eight countries (8.8) (P < 0.001). Venezuela also had lower rates for all five VATERL defects, even after excluding the 138 VATERL cases. VATERL cases were preferentially males (male proportion 0.6261) (P < 0.02), and, when compared with healthy controls, they had a higher perinatal mortality rate (63.7%) (P < 0.005), a higher frequency of fetal losses in previous pregnancies (12.6%) (P < 0.05), and lower mean birthweights (2,361.79 +/ 809.63 g) (P < 0.005). VATERL cases showed a higher rate than matched malformed controls for prenatal exposures to drugs and physical agents (P < 0.02 and P < 0.05, respectively), although no specific pharmacological or physical group was involved. The lower birth prevalence rates found in Venezuela, for VATERL as well as for each of the five congenital anomalies involved in this association, seem to be biologically meaningful. Since we could not identify a potential risk factor, nor a common cause of underascertainment unique to the Venezuelan subsample and common to all six hospitals, no hypothesis can be advanced here for this phenomenon. Nevertheless, this unequal geographic distribution strongly suggests a common etiopathogenicity for the five congenital anomalies involved in the VATERL association. PMID- 9409867 TI - Clinical and epidemiological characteristics of infants with body wall complex with and without limb deficiency. AB - The spectrum of defects in cases with limb body wall complex (LBWC) is quite variable since other anomalies are also observed in infants with LBWC, and some cases do not have limb deficiencies. Van Allen et al. [Am J Med Genet 1987;28:529 548] proposed that the diagnosis of LBWC (presence of body wall defects with evisceration of thoracic and/or abdominal organs, limb deficiency, and myelocystocele) should be based on the presence of two of three of the following anomalies: exencephaly or encephalocele with facial clefts, thoraco and/or abdominoschisis, and limb defects. This approach implies that an infant with encephalocele with facial clefts and limb defects may be considered as having LBWC, which I do not think is correct. I present the results of a clinical and epidemiological analysis aimed at identifying if, from an epidemiological perspective, it is possible to identify an entity which is characterized by the presence of abdominal wall defects along with other malformations including or not limb deficiencies. The result of this analysis allows us to consider that this entity should be characterized by the presence of abdominal wall defects with a variable spectrum of anomalies (with or without limb deficiencies) and, consequently, be called body wall complex (BWC). BWC includes cases regardless of their clinical pattern and the possible etiology or pathogenetic mechanism. Thus, the BWC entity does not include amniotic band sequence without body wall defects, but does include amniotic band sequence with body wall defects. PMID- 9409868 TI - Epidemiological characteristics of amniotic band sequence (ABS) and body wall complex (BWC): are they two different entities? AB - In 1965, Torpin [1965; Am J Obstet Gynecol 91:65-75] concluded that fetal constrictions and amputations were related to amniotic membrane rupture. Since then, this view was accepted widely, although different terms were used due to the variation in the types of associated anomalies. I consider that for those cases without body wall defects, the most appropriate term is amniotic band sequence (ABS), since amnion rupture could initiate a cascade of abnormalities (consequences) leading to the final structural defects. Here I present a clinical and epidemiological analysis of a series of consecutive infants with ABS, comparing their characteristics with those of infants with body wall complex (BWC). The results of this analysis suggest that ABS, from an epidemiological standpoint, is an entity different from BWC, and that amniotic bands in cases of BWC may be considered different from amniotic bands in cases without body wall defects. The former are probably produced earlier in the development than the latter. PMID- 9409869 TI - Cytogenetic and clinical findings in a patient with a deletion of 16q23.1: first report of bilateral cataracts and a 16q deletion. AB - The most commonly reported manifestations of 16q deletions are severe growth and developmental disorders and anomalies of the craniofacial, visceral, and musculoskeletal systems. We reviewed the findings of patients reported with 16q- syndrome and compared them to our patient, a 4 1/2-year-old boy with a deletion of 16q23.1. Findings include psychomotor retardation, hypotonia, high forehead, hypertelorism, upslanting palpebral fissures, low-set abnormally modeled ears, and talipes equinovarus. Anomalies present in our patient not reported in others with 16q- syndrome include bilateral cataracts, iris coloboma, and autistic-like behavior. It is of note that a locus for autosomal dominant congenital cataract, known as Marner cataract, was mapped previously to 16q22. Because our patient has bilateral cataracts and a unilateral iris coloboma, it seems likely that a gene involved in ocular development is located within 16q23.1. Our patient's deletion may also include the gene involved in Marner cataract and may further assist in the isolation of this gene. PMID- 9409870 TI - Possible new autosomal recessive syndrome of partial agenesis of the corpus callosum, pontine hypoplasia, focal white matter changes, hypotonia, mental retardation, and minor anomalies. AB - We describe a brother and sister with severe developmental delay, hypotonia, partial agenesis of the corpus callosum, pontine hypoplasia, focal white matter degenerative abnormalities, macrocrania, frontal bossing, deep-set eyes, and hypertelorism. The brother also had Duane syndrome type II and an ectopic right ureter. The coexistence of these multiple physical and brain abnormalities in a brother and sister suggests a new autosomal recessive syndrome with a slowly progressive course. PMID- 9409871 TI - Amelia: analysis of its epidemiological and clinical characteristics. AB - Eighteen cases of amelia in the Spanish Collaborative Study of Congenital Malformations (ECEMC) were analyzed epidemiologically. Prevalence at birth was 0.15 per 10,000 newborn infants, which is not different from that reported by other authors. Affected females outnumbered males. When compared with the control group, a lower birth weight, shorter gestation, lower placental weight, greater frequency of single umbilical artery, noncephalic presentation at birth, and more frequent maternal vaginal bleeding were observed in amelia cases. There were no significant variations of parental age. None of these patients was exposed to known teratogens, apart from 1 born to a diabetic mother; 3 patients had a genetic condition. Comparison of these variables with other studies is difficult because there is only one study that specifically analyzed amelia. Our data together with previous observations suggest that the genetic basis of amelia might be more important than has been considered previously. PMID- 9409872 TI - Poland sequence with dextrocardia: which comes first? AB - We report on two cases of Poland sequence (defect of the pectoralis major and hand on the same side) with dextrocardia, and review the literature on such patients. In all 16 reported cases, the Poland defect was on the left side, and associated with a rib defect, whereas most cases of Poland sequence involve the right side, and few have a rib defect. The dextrocardia appeared to be a dextroposition, and was not associated with other cardiac defects, whereas isolated dextrocardias (without situs inversus) frequently are. These observations suggest that the dextrocardia associated with Poland sequence is usually secondary to it. PMID- 9409873 TI - Familial neurofibromatosis 1 microdeletions: cosegregation with distinct facial phenotype and early onset of cutaneous neurofibromata. AB - A notable subset of the recent literature on the disorder neurofibromatosis type 1 (NF1) describes patients with NF1, facial anomalies, and other unusual findings. We describe a molecular re-evaluation of two such families reported previously by Kaplan and Rosenblatt [1985], who suggested that their NF1 manifestations, facial phenotype, and other findings could result from a disorder distinct from NF1. Submicroscopic deletions involving the NF1 gene were identified in both families by fluorescent in situ hybridization and analysis of somatic cell hybrids. Affected subjects of the first family were heterozygous for a microdeletion of approximately 2 Mb, which included the entire NF1 gene and flanking contiguous sequences. The family was remarkable for cosegregation of the NF1 microdeletion with facial abnormalities and a pattern of early onset of cutaneous neurofibromata upon transmission from an affected mother to her three affected children. The propositus of the second family carried a deletion that at the least involved NF1 exon 2 through intron 27, which is > 200 kilobases in length. Because all persons in the family were deceased, the size of the deletion could not be determined precisely. Facial anomalies were observed in the propositus and his NF1-affected mother and sister. The data from these families support our hypothesis, which was initially based solely on sporadic deletion cases, that deletion of the entire NF1 gene, or in conjunction with deletion of unknown contiguous genes, causes the facial anomalies and early onset of neurofibromata observed in this subset of NF1 patients. In addition, other features observed in the persons in these families suggest that some NF1 microdeletion patients may be at increased risk for connective tissue abnormalities and/or neoplasms. PMID- 9409874 TI - Severe limb deficiencies, vertebral hypersegmentation, and mirror polydactyly: two additional cases that expand the phenotype to a more generalized effect on blastogenesis. AB - We report on 2 unrelated fetuses with a multiple congenital anomaly pattern of severe limb deficiencies, vertebral/rib alterations, and mirror polydactyly similar to that described previously by us [Urioste et al., Hum Genet 97:214-217, 1996]. In addition, the two cases we present here have a more extense alteration of blastogenesis, expanding the phenotype of the cases previously reported. We have suggested [Urioste et al., Hum Genet 97:214-217, 1996] that this condition may be caused by a mutation in a developmental control gene that affects body plan organization. The minimal estimate of the prevalence of this new entity in our population (the Spanish Collaborative Study of Congenital Malformations, ECEMC) is 3.0/ 1,000,000 live births. PMID- 9409875 TI - Three brothers with mental and physical retardation, hydrocephalus, microcephaly, internal malformations, speech disorder, and facial anomalies: Mutchinick syndrome. AB - We describe three brothers from a non-consanguineous family with microcephaly, mental and physical retardation, speech disorder, facial anomalies, and internal hydrocephalus in two of the three affected brothers. The youngest brother died at the age of 5 months. He had situs abdominalis inversus, ASD II, and had been operated for internal hydrocephalus and atresia of the biliary duct. A search in the Oxford Medical Database Dysmorphology Program suggested phenotypic similarities with two sisters described in 1972 by Osvaldo Mutchinick in Argentina. Although differences in their phenotypes exist, it is possible that the two sets of sibs represent the same, rare syndrome. This interpretation is supported by the origin of both families from the same geographic region. PMID- 9409876 TI - Epidemiology of holoprosencephaly and phenotypic characteristics of affected children: New York State, 1984-1989. AB - Holoprosencephaly is a congenital defect of the median structures of the brain and face. The epidemiology is poorly known due to the paucity of population-based studies. This study describes the epidemiology of holoprosencephaly in a large population, using cases identified through the New York State Congenital Malformations Registry, and born in 1984-1989. We describe the craniofacial abnormalities present, their frequency, and their cooccurrence, and we examine the correspondence between the severity of craniofacial abnormalities, chromosomal abnormalities, and severity of the brain defect. Liveborn cases totaled 78, yielding a prevalence of 4.8 per 100,000 live births. Prevalence among girls was nearly double that in boys, and was 4.2 times higher among infants of mothers under age 18 compared to infants of older mothers. Only 9.8% of all cases had no craniofacial abnormalities other than the brain defect. Eye malformations were present in 76.8%, nose malformations in 69.5%, ear malformations in 50%, and oral clefts in 41.5%. These malformations arise at different times during gestation. The variability in patterns of cooccurrence suggests variability in the developmental pathways and/or timing of developmental derangements which result in holoprosencephaly. This, in turn, is consistent with a model of multiple causes. Children with alobar holoprosencephaly tended to have the most severe craniofacial anomalies, but the correspondence was not 100%. Craniofacial phenotype does not consistently discriminate between cytogenetically normal and abnormal cases. PMID- 9409877 TI - Progeroid syndrome with characteristic facial appearance and hand anomalies in father and son. AB - We report on the father-to-son transmission of a progeroid syndrome characterized by facial anomalies, sparse subcutaneous fat, and hand anomalies including syndactyly, camptodactyly, and finger deviation. Mild mental retardation, microcephaly, and congenital heart defect were found only in the son. To our knowledge, this syndrome has not been described previously. PMID- 9409878 TI - Magnetic resonance imaging abnormalities of the brain in Goldberg-Shprintzen syndrome (Hirschsprung disease, microcephaly, and iris coloboma) PMID- 9409879 TI - Trichorhinophalangeal syndrome type 2: another syndromic form of hydrometrocolpos. PMID- 9409880 TI - [Special issue on feto-placental pathology]. PMID- 9409881 TI - [The feto-placental examination and public health]. PMID- 9409882 TI - [Role of the pathologist in prenatal diagnosis and genetic counselling]. PMID- 9409883 TI - [Prenatal diagnosis of renal malformations and diseases]. PMID- 9409884 TI - [The pathologist confronted with hydrops fetalis. Management and principal etiologies]. PMID- 9409885 TI - [Fetoplacental infections]. PMID- 9409886 TI - [Placental examination and intrauterine growth retardation]. PMID- 9409887 TI - [Detection of chromosome anomalies using in situ hybridization on fetal tissue sections]. AB - The use of in situ hybridization to detect and characterize chromosome anomalies on cytogenetic preparations is now largely applied in clinical laboratories. Its use in embryofetopathology on formalin-fixed and paraffin-embedded tissues, when an aneuploid condition is suspected but a routine chromosomal analysis is not possible was assessed in the present study. Control values of hybridization signals obtained after different enzymatic digestion protocols have been established on normal fetal tissues. Tissue conservation and fetal age as well as other parameters have also been analysed. A successful hybridization has been achieved in most cases. Failure of hybridization was observed in specimens associated with extensive tissular lysis. In conclusion, the application of in situ hybridization on fetal tissues is very useful to detect chromosome anomalies in embryofetopathology. PMID- 9409888 TI - [Diagnosis of lysosomal storage diseases with fetal presentation]. AB - In metabolic diseases with fetal presentation, lysosomal storage disorders represent a fairly homogeneous group. Hydrops fetalis or ascites is the main but non specific symptom. The relative frequency of lysosomal storage diseases in this context is not well known and probably underestimated. They represent 1.4% in a large retrospective series. The contribution of the placental and fetal examination in their diagnosis is emphasized. The biological investigations required for their accurate pre and postnatal diagnosis are described. The precise identification of the lysosomal defect is necessary to propose an early prenatal diagnosis by chorionic villi biopsy in further pregnancies. PMID- 9409890 TI - [Diagnosis of fetal akinesia except for oligoamnios]. AB - The term Fetal Akinesia Sequence (FAS) covers a large spectrum of developmental abnormalities resulting from a lack of intra-uterine fetal movements, which share heterogeneous etiologies. Environmental, "extrinsic" causes are easily ruled out. Various neuromuscular disorders, involving the motor unit at any level, constitute the main part of "intrinsic" fetal pathology. We propose a detailed schedule of prospective investigation of FAS, in order to standardize and gather the most pertinent information and to compare a wide panel of accurate data between fetopathological centers. The objective is to improve the understanding of various pathogenetic processes involved in the emergence of FAS, in order to propose better information and genetic counselling to parents, and potentially, to consider a prenatal prevention. PMID- 9409889 TI - [Fetal neuropathology: a new approach]. AB - Recognition of central nervous system (CNS) abnormalities by prenatal imaging has awakened interest in fetal neuropathology, considered as a terra incognita by most pathologists. The evaluation of fetal brains implies a perfect knowledge of the timing and characteristics of the developing and changing nervous system during gestation. Disorders of the brain are now studied with special regard toward their cause analysis considering fetal, maternal, and placental pathology. Thus, with this new approach and the distinction between primary and secondary CNS lesions, an effective genetic counselling has become a realistic goal. Examining fetal brains has led on the other land, to the recognition of distinct entities and the identification of specific chromosomal and/or gene(s) abnormalities. PMID- 9409891 TI - [How to research oligohydramnios?]. AB - When an oligohydramnios has been clinically detected, the fetoplacental study has to find the cause of this syndrome: malformations, infectious or environmental anomalies, and to study the consequences of this syndrome in the fetus. A careful and complete study allows us to indicate some genic syndromes such as polycystic kidney disease, Meckel syndrome, familial renal tubular dysgenesis, Vacterl Hydrocephaly sequence, or Fraser syndrome inducing the genetic counseling and a familial study. The other pathologies, such as maternal diseases, infections or sporadic malformations must be followed by cautious maternal care for subsequent pregnancies. PMID- 9409892 TI - [Diagnosis of a fetal cardiac malformation. Fetal heart]. AB - To diagnose a fetal heart abnormality is one of the most important purpose of the foetopathologic examination performed after spontaneous abortion, intrauterine death or pregnancy termination. Frequency of cardiac abnormalities in fetal or maternal pathologies explains the significance of this diagnosis in a pertinent genetic counseling before next pregnancy. The diagnose of a fetal heart abnormality requires a good knowledge of: normal anatomy and examination technics of fetal heart, congenital heart abnormalities classification and knowledge of the most common diagnosis, diagnostic orientation according to the cardiopathy: fetal origin in genic syndrome, chromosomal abnormalities, malformative or sporadic association; maternal etiology (toxic, infection, metabolic). This procedure needs experience, acquired by rigorous practice and team working. PMID- 9409893 TI - [Diagnosis of meconium ileus]. AB - Meconium ileus represents a functional disorder mainly detected during the second trimester of gestation. The diagnosis can be recognized by bowel hyperechogenicity or by the pathologist after a systematic examination. It must lead to etiologic diagnosis and specific investigations according to the history. Even in absence of family history, cystic fibrosis must be considered with pancreatic study and with frozen fetal material in order to be able to search for the most common mutations. Meconium ileus can also be associated with infection, fetal blood swallowing, chromosomal aberrations and multiple fetal abnormalities. Only good investigations for specific diagnosis can lead to a genetic counsel and a good follow up of the next gestation. PMID- 9409894 TI - 5th European Meeting of Environmental Hygiene. Prague, Czech Republic, June 27 29, 1995. Abstracts. PMID- 9409895 TI - [Analysis and conclusions regarding the epidemic spread of methicillin resistant S. aureus]. AB - Increasing dissemination of Methicillin-resistant S. aureus (MRSA) calls for a more consequent examination of the causal mechanisms. Experiences and results of analysis of three different outbreaks caused by MRSA are described. In summary it is shown that MRSA are especially epidemic virulent and persist over a long range of time in a care unit. Main reasons for these outbreaks obviously were mistakes in hospital hygiene and an inappropriate antibiotic prophylaxis. Predisposed inpatients suffer from severe infections. Interregional dissemination of MRSA observed is mainly due to the transfer of patients between hospitals. Optimization of hospital hygiene especially isolated or cohort nursing of affected patients as well as restriction of antibiotic prophylaxis and therapy are crucial measures against the dissemination of MRSA. For success of these measures information and motivation of staff are necessary. PMID- 9409896 TI - [Hygienic processing of dental transmission instruments (handpieces and instruments, turbines) in dental practice]. AB - The efficacy of a steam disinfection method for the processing of used instruments in dental practice was tested with 274 high-speed turbines and handpieces. The instruments treated with streaming steam were both taken out of the dental practice of treatment and artificially contaminated with five different test-microorganisms (E. coli, P. aeruginosa, S. aureus, E. faecium, C. albicans) and different concentrations of serum (1%, 10%, 20%). In spite of the narrow channels of these instruments a microbial reduction of more than 5 log10 steps could be achieved in all cases. Thus dental high-speed turbines and handpieces can reliably be disinfected with streaming steam. PMID- 9409897 TI - [Comparison of perforated metal ceiling systems (supported airflow ceilings) with laminar airflow ceilings in type A (DIN 1946 T.4) operating rooms under surgical conditions]. AB - In eleven centrally ventilated operating theatres the concentration of particles and airborne germs in wound vicinity was measured on three workdays. Five theatres were equipped with air supply ceilings with supporting flow outlets (supporting flow ceilings), five with laminar air flow ceilings and one with an air supply ceiling, a body exhaust system and a partition wall between the anesthetic and operating areas. Under routine conditions the air supply of the laminar air flow ceiling with its lower turbulence shielded the operating field from the largely staff-related air contamination in the rest of the theatre better than in the case of the supporting flow ceilings. Particles and airborne germs were removed from the endangered wound area faster. A spatial separation between the anesthetic and the operating areas as well as a body exhaust system lead to a considerable reduction of the contamination. Two theatres were conspicuous by reason of their considerably raised values due to defective control engineering and the wrongly positioning of the operating table. From the point of view of ventilation technique the laminar air flow ceilings with lower turbulence are superior to air supply ceilings with supporting flow outlets in the working day of an operating theatre. In order to minimize the influence of the staff, which up till now has been neglected in testing specifications, constructional possibilities such as the size of ceiling, the partitioning off of operating and anaesthetic areas and the positioning of the operating table in relation to the incoming air should be coordinated rationally. Taking measurements regularly during operations can provide the impulse for considerable improvements in both operational and planning phases. PMID- 9409898 TI - [New data on syncarcinogenesis in tumors of exogenous origin]. AB - Little is known about syncarcinogenic effects of occupational and environmental substances although it is supposed that different exogenous factors may play critical roles in the development of many human tumors. Epidemiologic results prove syncarcinogenesis for asbestos exposure and smoking (lung cancer), radon exposure and smoking (lung cancer), exposure to aromatic amines and smoking (bladder cancer) and alcohol abuse and smoking (oral, larynx and oesophagus cancer). Animal experiments point to additive effects in carcinogenesis for different nitrosamines and substances like benzo(a)pyrene, carbon tetrachloride, ethanol, vinyl chloride and ionising radiation. It can be concluded from modern concepts of carcinogenesis that syncarcinogenic mechanisms may not only result from genotoxicity but also from influences on cell proliferation and mitogenesis as well as toxicokinetics, DNA repair, intercellular communication, immune system and hormonal effects. New methods of molecular epidemiology seem very promising to study syncarcinogenic effects in animals and humans. PMID- 9409899 TI - [Mutagenicity, genotoxicity and cogenotoxicity of environmentally relevant nitro musk compounds]. AB - In the present study a new in vivo/in vitro animal model was used to study the ability and potency of musk ketone and musk xylene to induce liver specific oxygenases (in vivo) which are necessary of toxify different premutagens, pregenotoxicants and/or precarcinogens to the ultimate DNA damaging agents. Therefore, rats were pretreated with 10, 20 and 40 mg/d nitro musk (NMV) for 5 days by intraperitoneal (i.p.) injection. Then the postmitochondrial fractions of the hepatocytes (S9M) were used to examine the metabolic potency for toxification of the pregenotoxicants benzo[a]pyrene (B[a]P) and 2-aminoanthracene (2-AA) using the SOS chromotest (in vitro). Furthermore, musk xylene, musk ketone, musk ambrette, musk moskene and musk tibetene were examined for their mutagenicity in the Salmonella/microsome assay using S. typhimurium TA97, TA98, TA100 and TA102 and for their genotoxicity in the SOS chromotest using Escherichia coli PQ37 (sfiA::lacZ) in the presence and absence of an exogenous metabolizing system (S9 of PCB induced rats = S9A). Both musk ketone and musk xylene were identified als inducers of toxifying enzymes (oxygenases) in rat liver. Using the in vivo/in vitro model these isoenzyme inductions led to a metabolisation (toxification) of the pregenotoxicants benzo[a]pyrene (B[a]P) and/or 2-aminoanthracene (2-AA) (cogenotoxicity). Using S9M fractions of rats which were i.p.-pretreated with 5 x 40 mg musk ketone the induction factor in the SOS chromotest was IFmax = 4.0 by using 1 nmole B[a]P and IFmax > 4.0 by using 20 nmole 2-AA. Thus, musk ketone seems to be a Cytochrome P450 1A1 and 1A2 isoenzyme inducer in mammals. On the other hand the S9M fractions of musk xylene pretreated rats showed only a toxification of 2-AA (IFmax = 3.0). Therefore, a synergistic effect of enzyme inducers, i.e. musk xylene and musk ketone, and pregenotoxicants, i.e. B[a]P and 2-AA, regarding DNS damaging effects was identified. Musk ambrette showed high mutagenicity in S. typhimurium TA100 (500 His(-)-revertants per mumole, +S9A). Unexpectedly, these DNA damaging effects were not caused by bacterial nitroreductases but by rat S9A metabolisation (!). SOS inducing DNA damages in E. coli PQ37 were not produced (IFmax < 1.5). On the basis of the results presented and under consideration of the concentrations of NMV, other cogenotoxicants and pregenotoxicants such as B[a]P and 2-AA in environmental samples and human tissues, a genotoxic risk for humans has to be assumed. PMID- 9409900 TI - [Gas chromatography and mass spectrometry method for detection of selected pyrethroid metabolites in urine]. AB - Pyrethroids are increasingly used indoors, because of their low mammalian toxicity but high insecticidal activity. Indoor application of pyrethroids, which is often carried out without adequate expert knowledge, may lead to high pesticide residues, often combined with health hazards for the people concerned. Objective of the present study is the development of a method for a biological monitoring to determine internal exposure to pyrethroids. Major pyrethroid metabolites were detected in urine by capillary gas chromatography in combination with mass selective detection (MSD). For the metabolites, the limits of determination range between 0.5 and 1 microgram/L urine. Our results so far demonstrate the detectability of cis- and trans-3-(2,2-dichlorovinyl)-2,2 dimethylcyclopropanecarboxylic acid (DVCA) and 3-phenoxybenzoic acid (3-PBA) in urine during a period of elimination of 48 hours after exposure to cypermethrin. In this paper the method is exemplary illustrated by one exposed person (pest control operator). PMID- 9409901 TI - [Serologic studies of domestic cats for potential human pathogenic virus infections from wild rodents]. AB - For several viral infections a reservoir in wild rodents has been demonstrated. Some of the agents are known or suspected to be pathogenic for humans. Because improvements in hygiene have reduced direct human contact with rodents, domestic cats could be acting as active transmitters of these viruses from rodents to man. We selected 4 such pathogens--ortho- and parapox-, hanta- and encephalomyocarditis viruses--which, in different ways, may lead to serious human illness: Ortho- and parapoxvirus infections may cause localized pox lesions following direct skin contact. In general, the lesions heal without complications; in immunosuppressed or -deficient individuals, however, infection may generalize and take a dramatic course. Hantaviruses exist in various serotypes with different pathogenicity for human beings, varying from asymptomatic infection to highly fatal disease. In central and northern Europe the Puumala serotype is predominant causing influenza-like symptoms and renal dysfunction. Human infections arise from inhalation of aerosolized excreta of persistently infected rodents. Infections of man associated with encephalomyocarditis virus were demonstrated sporadically in cases of encephalitis and meningitis. In the present study, we investigated in 200 feline serum samples the prevalence of antibodies to ortho- and parapox-, hanta- and encephalomyocarditis virus. All serum samples were from cats that had been allowed to roam outside and to hunt. They were submitted from all parts of Austria for routine diagnosis in 1993. Four per cent of cats showed antibodies to orthopoxviruses with haemagglutination inhibition (HI) titres of 16-512; because of extensive cross-reactivity, positive samples reacted with all investigated orthopoxviruses (a feline orthopoxvirus recently isolated in Vienna, the reference strain of cowpox virus, Brighton, and vaccinia virus, strain IHD), only varying in titre. The specificity of the results was confirmed by virus neutralisation (VN) test, in which the same sera showed titres of 4-32. These data imply that, at least in Austria, unrecognized or subclinical orthopoxvirus infection in cats is more common than previously thought. In contrast to orthopoxviruses, all serum samples proved negative to parapoxvirus (parapoxvirus bovis 1) in VN test. In the same 200 samples, a seroprevalence of 5% was found to hantavirus (immunofluorescence antibody assay), indicating that domestic cats are susceptible to this virus and that infection is not uncommon in cat populations. Because higher titres were obtained against the Puumala serotype compared to the more pathogenic serotype Hantaan, it is most likely that the cats had experienced Puumala infections. Using HI test, antibodies to encephalomyocarditis virus (EMCV) were demonstrated in only 1.5% of the feline serum samples; although the antibody titres were low (16 and 32, respectively) we consider them specific, because these sera proved positive in VN test as well. Nevertheless, EMCV infection in domestic cats seems to be of low importance. The serological results presented in this paper, together with virological and epidemiological data, indicate that the domestic cat plays an important role only in the transmission of orthopoxviruses to human beings, but not in the case of parapox-, hanta-, and encephalomyocarditis virus. PMID- 9409902 TI - [Nitrite formation and nitrosation potential of heterotrophic biological denitrification of drinking water in laboratory conditions]. AB - In a laboratory construction for heterotrophic biological denitrification of drinking water treatment, the formation of nitrite, the potency of nitrosation and the genotoxic activity were tested. Parameter as nitrate concentration, the water flow rate in the system, nitrite and morpholine addition and the pH-value were checked. For testing the potency of nitrosation and formation of nitrite in the reactor we took morpholine, a fast nitrosing amine. The results show for the break down rate of nitrate, there is no influence of the initial nitrate concentration (80-195 mg/L), the nitrite addition (5-20 mg/L) and the water flow rate (45-100 min) in the system. pH-values below 5.5 showed a little break down rate of nitrate. There was no correlation between the starting point of nitrate concentration and the formation of nitrite, although there was a positive correlation between the length of stay and the formation of nitrite. Nitrite concentrations of 5 mg/L with morpholine concentrations of 5 and 10 mg/L didn't show detectable formation of nitrosomorpholine. The analyses of different watertests in the construction didn't show significant results for DNA damage by the sister-chromatid exchange (SCE). The results of the Salmonella microsome assay (tester strain TA1535) didn't show any mutagenic effects relating to the potency of nitrosation. According to our experiments the potency of generalising nitrosamides or nitrosamines by drinking water denitrification seems to be low. There is no final assessment of detriment to health by denitrifying drinking water. PMID- 9409903 TI - [Effectiveness of peroxide solutions against microorganisms in biofilms]. AB - Central dosing units for surface disinfectants can constitute a particular hygiene problem in hospitals if a biofilm can develop in the piping system despite the disinfectant solution that is present. In earlier studies it was able to be shown that oxygen-releasing compounds in particular were highly effective against microorganisms in generated biofilms. The purpose of the study presented was to investigate in a test simulating practical conditions whether, in addition to the described good effect, peroxide-containing solutions also have a uniform effect against all isolates from disinfectant use-solutions, or whether there are differences. The results show that a 1% H2O2 solution (C) was not sufficiently effective against all test organisms after a contact time of one hour. In contrast, the 1% test solution of A (10% perglutaric acid, 28% H2O2, < 0.5% perbenzoic acid) was highly and uniformly effective. The 3% test solution of B (10% tertiary butylhydroperoxide, 20% phenoxypropanols, 48% dipropylene glycol) exhibited weaknesses against some test organisms after the one-hour contact time. After a contact time of three hours, this solution, unlike the 1% H2O2 solution achieved a reduction in bacterial count of more than 5 log steps against all species. On the basis of these results, before cleaning piping systems it appears advisable to test the effectiveness of the solution to be used in a suitable test with the isolates. PMID- 9409904 TI - [Effect of silver and copper ions on survival of Legionella pneumophila in tap water]. AB - In vitro studies were performed to give information about the required metal concentrations in decontaminating Legionella-loaded warm water systems with the electrochemical generation of Ag+ and Cu2+ ions. The influence of Ag and Cu ions, as single compounds and in combination, on the survival of Legionella pneumophila (serogroup 6) was determined in tap water at 45 degrees C. Marked differences were detected in the action of these metals. Ag produced a much stronger inhibition than Cu. No additive effect was demonstrated when using Ag/Cu combinations in the ratio of 1:10. In this case only the Ag-induced inhibition was detected. After 1 h of incubation at 45 degrees C a concentration of 80 + 800 micrograms/L Ag + Cu was needed to produce the maximal inhibitory effect (a 5 log decrease). An identical effect was seen after exposure to 20 + 200 micrograms/L Ag + Cu in the long-term action (24 h of incubation). The minimum inhibitory concentration after long-term incubation was 5 + 50 micrograms/L Ag + Cu. These metal concentrations produced a 1 log reduction. The in vitro results are discussed under consideration of earlier investigations after metering Ag and Cu into a Legionella-loaded water system and generated the following conclusions: In the beginning highly contaminated water systems at 45 degrees C need concentrations between 40 and 80 micrograms/L Ag + 400 to 800 micrograms/L Cu to kill Legionellas. After effective reduction of Legionella concentration of at least some logarithmic powers a slow constant maintenance concentration of 5 to 20 micrograms/L Ag + 50 to 200 micrograms/L Cu could be applied. At 22 degrees C the in vitro inactivation response is much lower. On the other hand in warm water systems with temperatures of 50 to 60 degrees C lower metal concentrations are sufficient. PMID- 9409905 TI - [Dust and microorganism count at delivery, sorting and composting of home refuse and home refuse-like industrial waste]. AB - The exposure of employees to airborne dust and microorganisms was assessed in a waste processing plant established to recover reusable materials from unsorted domestic and industrial waste. Exposure criteria considered relevant were the quantity of the individual size-selected particle fractions, the morphological properties of the particles, their heavy metal content, and the degree of their contamination with various microorganisms and mold. In addition, separate microbiological analyses to determine potential pathogen concentrations in the air were made. The highest concentrations of total and fine dust were measured in the waste delivery area. A close correlation between the frequency of deliveries and the level of dust exposure was observed. In this area, fine dust concentrations exceeded the threshold limit value of 6 mg/m3 repeatedly for shorter periods. The average fine dust concentration during an entire work-shift, however, was considerably lower than this value. Particles with an aerodynamic diameter of 2 to 7 microns predominated both in the waste delivery and the processing areas. Fibrous dust particles were present in smaller numbers than spherical particles and consisted mainly of organic materials. Natural and artificial inorganic fibers were found only occasionally. The concentrations of lead, cadmium, nickel and mercury were considerably lower than the corresponding MAK and TRK values, and were--with the exception of lead--in the range of the respective metal concentrations in the atmosphere of urban areas. Microscopical examinations of used protective masks (protection category P2) revealed that dust particles were deposited even on the inner side of the masks. Most of the particles on this side were very small and carried nickel or titanium. Microorganism concentrations measured in air from the highly dust-exposed areas of the plant showed values up to 6.9 x 10(5) cfu/m3, with a mold content of 6.6 x 10(4) cfu/m3. Approximately 90% of the microorganisms were deposited on particles of the fine dust fraction (particle size < 7 microns), and more than 50% contaminated particles with an aerodynamic diameter of 2 to 4.7 microns. In the compost facility, mold concentrations of up to 8.4 x 10(5) cfu/m3 were measured. In contrast, the level of microbial contamination in the filtered air from the compost facility did not exceed the concentration measured in air outside the plant. The data which were obtained during the winter months are probably at the lower end of the average exposure range over the entire year. In regard to exposure assessment, it should be mentioned that particles with a size of 4 to 7 microns are not really "inert" particles, since they are preferential carriers for heavy metals and microorganisms. More studies in the future should be performed to establish, whether the level of exposure to microorganisms can be estimated indirectly by the determination of dust exposure. PMID- 9409906 TI - [Environmental and indoor air exposure to asbestos fiber dust as a risk and causal factor of diffuse malignant pleural mesothelioma]. AB - In an interdisciplinary, multicentre case control study of the causal factors of the diffuse malignant mesothelioma (DMM) standardised histories where taken from n = 324 Patients suffering from DMM, n = 315 hospital control patients (KK) and n = 182 population controls (PK). For 66 DMM, 149 KK and 107 PK a risk from asbestos fibre dust at the workplace was not detectable. For latter persons indoor and outdoor asbestos exposure outside of the workplace were investigated. The following factors were examined: neighbourhood exposure from companies using asbestos, living in big cities and nearby main traffic roads, building materials containing asbestos, electric storage heaters and household contacts. For using electric storage heaters a statistically significant increased odds ratio (OR) was observed for DMM as well in comparison with KK (OR = 2.42; 95%-CI: 1.01-5.72) and in comparison for PK (OR = 2.91; 95%-CI: 1.08-7.80). Only outside of Hamburg an increased OR compared to KK was observed for people living in the neighbourhood of asbestos factories (OR = 16.3; 95%-CI: 1.35-196.8) and also, but only in Hamburg, compared to PK living nearby main traffic roads. There is only a trend for a mesothelioma-risk for household-contacts based on a few cases. In one DMM-patient without an occupational asbestos exposure the lung dust fibre analysis yielded 2.912 FB and 1.459 x 10(3) crocydolithe fibres per gram dried lung tissue. As a child he lived in the immediate vicinity of the blue asbestos mine in Wittenoom, Australia. Therefore in special cases a para-occupational asbestos or a neighbourhood asbestos exposure can be demonstrated as a risk factor of diffuse malignant mesothelioma. PMID- 9409907 TI - [Internal hazardous substance burden of persons from various regions of origin- studies of lead, mercury, arsenic and cadmium exposure]. AB - AIM OF THE STUDY: The aim of this study was to investigate the concentration of metals of environmental-medical relevance in biological materials in persons seeking asylum with regard to their country of origin. COLLECTIVE AND METHOD: During medical examination after entry into Germany of persons seeking asylum, samples were taken for determination of the following biological monitoring parameters: lead in blood, and arsenic, cadmium and mercury in urine. A total of 103 males were investigated (13 from former Yugoslavia, 29 from the former USSR, 33 Africans and 28 Asians) ranging from 16 to 53 years of age (median 27 years). 34 male Germans without occupational exposure to these substances and a similar age structure (age 25-36 years; median 26 years) served as a control group. RESULTS: The countries of origin had a significant influence on all the biological monitoring parameters investigated. The mean blood lead concentration in the Asians of 75.4 micrograms/L was the highest level found, while the lowest concentration of 38.0 micrograms/L was measured in the German controls. Also the level of arsenic excreted in the urine was on average much higher in the persons seeking asylum than in the German controls. In the Africans a mean level of 9.7 micrograms/g creatinine was reached. The Germans had the lowest arsenic concentrations in urine of 5.3 micrograms/g creatinine. There were, however, considerable interindividual fluctuations, which are probably due to oral uptake of arsenic compounds as a result of eating seafoods. The highest mean concentration of mercury excreted in urine was found in the German controls. Values of 0.9 microgram/g creatinine were determined. The men seeking asylum from former Yugoslavia had significantly higher values than other groups for cadmium excreted in urine. The median of 0.6 microgram/g creatinine was nearly three times as high as found in the Germans. CONCLUSIONS: For all parameters investigated, with the exception of mercury, higher internal exposure was found in the persons seeking asylum than in the German controls. This may be due to individual life style, dietary habits or environmental conditions in the country of origin. For clinical environmental medicine, the 95th percentile, as the upper limit of the reference range, can only be regarded as an orientation aid for classifying the exposure to hazardous substances of an individual compared to other persons from the same environment. PMID- 9409908 TI - [Emission of odors from composting of biological waste]. AB - Results of qualitative and quantitative detection of odour substances during the process of composting of biowaste are shown. Olfactometric measurements were carried out beside detection of odour substances by gaschromatography--mass spectroscopy. Some odour relevant substances were set in relation to odorant concentration. 158 volatile substances could be identified by GC/MS. The occurrence of odour substances demonstrated a characteristic dynamic. As a consequence of analysis of odour substances, detection of odorant concentration and odour quality the composting process could be divided in 3 main phases. For the start phase substances from anaerobic degradation like alcoholes and esters of carbonic acids played an important role in odour generation. Sulfur containing compounds dominated the characteristic odour during thermophilic phase of composting process. The importance of ammonia during later composting process could be shown. The concentration of dimethyl disulfide and limonene was set in relation to odorant concentration and both substances are discussed as indicator compounds. PMID- 9409909 TI - [Influence of chewing gum consumption and dental contact of amalgam fillings to different metal restorations on urine mercury content]. AB - It had been shown previously by various authors that contact of amalgam fillings to metal fillings of different type can increase the electrochemically caused amalgam corrosion in vitro thus leading to an elevated release of mercury. So it was recommended to renounce of a dental contact of amalgam to metal fillings of other type. One aim of the present study was to evaluate possible influences of this contact in vivo on the urinary mercury contents in human volunteers. Neither approximal nor occlusal contacts had any influence on the urinary mercury excretion in comparison to a reference group with similar amalgam status. Furthermore, the influence of gum chewing on urinary mercury levels was taken into account. It could be shown that the consumption of chewing gum resulted in a significantly higher mean urinary mercury content in probands with amalgam fillings in comparison to people with similar amalgam status (gum chewers: 1.36 Hg/24 h vs. non-chewers 0.70 microgram Hg/24 h). Thus, gum chewing has to be considered as important parameter of influence on the urinary mercury levels of people with amalgam fillings. PMID- 9409910 TI - [Colony blot method for detection of legionellas--results of a comparative study]. AB - In the following short communication a new commercially available immunoassay for the quantitative detection of Legionellae after cultivation was compared with the conventional method recommended by ISO in a study shared by 6 laboratories. After a training phase of the laboratory personal a very good agreement of immunological and conventional method was observed by testing 310 water samples. The colony blot assay for quantitative identification of Legionella spec. is a rapid and specific method and can be recommended for quantification of Legionella spec. in water samples. PMID- 9409911 TI - Proceedings of the 14th Dusseldorf Hygiene Conference. April 24-25, 1996. PMID- 9409912 TI - [Legal requirements for health protection from the European viewpoint--uniform regulations or reciprocal recognition of public health norms?]. AB - Are uniform legislative measures and complete harmonization of i.a. sanitary standards required or is the application of the principle of mutual recognition to be preferred when it comes to ensure the protection of public health in the context of the creation and further development of the single market? As a matter of fact, we can neither ensure the proper functioning of an internal market without technical barriers nor satisfy the necessary requirements of health protection without having recourse to legislative measures. However, the adoption and implementation of community legislation does imply--as we have seen in the past--an extensive harmonization of technical standards which in turn may involve the dual risk of a harmonization of health protection standards at the lowest level and/or, in certain cases, of an "over-implementation" of the Community standards in the Member States. In order to counter this dual risk, a more flexible approach has been adopted in various sectors. Yet, a decentralized implementation of the community measures does undeniably involve a certain risk of distorting competition within the single market. Two groups of experts, known as the Sutherland-Group and the Molitor Group, have dealt with these issues at some length in two reports, which were published in October 1992 and in June 1995 respectively. In the light of these reports and on the basis of the conclusions of the European Council in Edinburgh (December 1992) and the so-called "Brussels Programme" of the European Commission (December 1993), a certain number of measures have now been taken with a view to overcome the above-mentioned problems. Thus, framework directives are now more generally applied instead of detailed legislative measures. Moreover, as a sort of alternative to further legislation, certain measures have been proposed in order to ensure the proper operation of the principle of mutual recognition and thus a less cumbersome management of the single market. The creation of an EU-wide network of independent "consumer ombudsmen" could contribute substantially to ensure the implementation of health and safety standards throughout the European Union. And last, but not least, legal sanctions must be imposed on those Member States that do not comply with the community legislation to the detriment of the health and safety of the EU citizens. PMID- 9409913 TI - [Risk assessment and risk management according to the HACCP (Hazard Analysis and Critical Control Point) concept: a concept for safe foods]. AB - The volume of official hygiene regulations for food processing establishments has been growing continuously over the past 20 to 30 years. This led to a decrease in hygiene risk awareness in food processing establishments which was partly replaced by a strong reliance on legislative measures in food hygiene. After experiences in industrialized nations had shown that even numerous and detailed hygiene regulations could not prevent the increase of infections and intoxications of consumers by food products, new solutions had to be found. On the one hand, the implementation of intensified control measures by the producers themselves is required. Such a control much then be "controlled" by the state authorities. On the other hand, the so-called HACCP (Hazard Analysis and Critical Control Point) system has been introduced as a new quality assurance principle for the avoidance of health hazards. This concept was developed in the 1960s in the United States of America in order to produce safe foods for the space programme. For the production of a particular food according to the HACCP system informations on hazards and situations leading to their presence are collected and evaluated in order to decide which are significant for food safety and therefore should be addressed in the HACCP plan. According to this analysis the necessary preventive measures which lead to the prevention, elimination or reduction to an acceptable level of identified health hazards have to be defined. All steps of food processing have to be included in the HACCP system. Raw materials, storage of foods, types of distribution and the intended usage of the final product by the consumer have to be considered in this system. However, the introduction of the HACCP system into European hygiene regulations does not constitute an entirely new development. It can rather be regarded as a renaissance of traditional scientific concepts. This is demonstrated by the example of drinking milk processing as it was practised sixty years ago. PMID- 9409914 TI - [Quality assurance in food production in Europe according to ISO 90000 and HACCP (Hazard Analysis and Critical Control Points)]. AB - HACCP is intended to make food protection programs evolve from a mainly retrospective quality control toward a preventative quality assurance approach and to provide an increased confidence in food safety. In parallel, the continuous evolution of quality concepts in the food industry resulted in the development of quality systems and quality assurance techniques with regard to the EN 29,000 (ISO 9000) series of standards. Specific to the food industry, HACCP can be seen as a very effective method to prepare specific Safety Assurance Plans (cf. the quality assurance plan concept) within a quality systems approach (Jouve, 1993). By reference to ISO 8402, a Quality Assurance Plan (QAP) basically sets out "the specific quality practices, resources and sequence of activities relevant to a particular product, service, contract or project". Quality assurance plans are more particularly useful for projects relating to new products or processes and/or comprising inter-related tasks whose interaction may be complex. In addition, in contractual or regulatory situations, such plans can be used to demonstrate the supplier's capability to meet identified objectives, specification or standards. In the food industry, the management of safety is a critical and complex issue which fits very well in the scope of application of a specific QAP; it is also where the use of HACCP is otherwise recommended by priority. PMID- 9409915 TI - [Quality management and quality assurance in the hospital with special reference to hospital public health]. AB - Due to the law of health reform and health structure the hospitals in the Federal Republic of Germany are asked to not only think about cost reduction but at the same time to also face the requirements of quality management (QM) and quality assurance (QA). For the field of medical and care taking treatment of patients in the hospital the term "quality" can be described as the total of characteristics of the medical services necessary for the realization of all requirements which lead to an optimal patient treatment. Hospital hygiene as part of hygiene in the broader sense deals with the bases of disease prevention of all persons actively or passively connected with the institution hospital as well as with the natural and social hospital environment and, thus, in every respect proves to be a guide discipline for the realization of QM and QA in the hospital. For the realization of an overall concept for QM and QA in the hospital three categories of quality have proved to be worthwhile, namely the structure quality which deals with the quality related arrangement of the frame conditions of hospital productivity the process quality which comprises the improvement of diagnostic, therapeutic, care taking and medical activities in the hospital as well as interactions between the hospital staff and the patient or his relatives, and the outcome quality, which judges the changes in the patient's condition by means of predefined target values. Furthermore, QM in the hospital needs detailed informative activities (initial stage of information) as well as motivating measures (initial stage of conduct) whereas latter has to be considered as an essential basis for a staff related realization of quality assurance measures in the hospital. Possibilities for a practical realization of QM and QA are shown by means of the modular concept established in the Clinics of the University of Heidelberg. Thereby, one could experience that a successful QM in the hospital requires the declared willingness of the hospital leadership to regard quality as the main theme of medical services and that QM has to be carried out staff-orientated for only by improvement of the employers' activities on all levels of the hospital gains the medical process of service its quality. PMID- 9409916 TI - [Status of public health in hospitals in France]. AB - The situation of hygiene in hospitals in France has been greatly improved in the late 1980's with the commitment for public hospitals to set up a local committee against nosocomial infections (N.I.). This has been completed by the creation of Coordination Committees at the regional level in France and one central National Committee in charge of defining politics and priorities in hygiene. From early 1990, several epidemiological surveys have been set up, both at the national and regional level, displaying nosocomial infection prevalence rates of approximately 6 to 16%. High risk groups population for nosocomial infections had been targeted as a priority in the different survey programs: wounds, bacteremias, surgery wards and intensive care units. The epidemiologic situation of France, as in south Europe, is characterized by a high rate of multiple antibiotic-resistant strains. The methicillin resistance rate of S. aureus was over 40%. A specific program to prevent Staphylococcus aureus transmission had led to a significant decrease of this prevalence rate (32%). Prevention of nosocomial infections is now considered by French Health Authorities and health care providers as a public health priority giving their frequency, morbidity and mortality rates, and costs. Optimizing prevention of nosocomial infections should be considered as a significant marker for quality of care and safety of hospitalized patients. They might play a significant role in budgeting hospitals. PMID- 9409917 TI - [Risk assessment and risk prevention in infectious diseases]. AB - The purpose of risk assessment is to provide source data to state and local health agencies and other administrative departments to aid in the development of approaches for risk management and preventive strategies. In respect of the far reaching consequences it is necessary to have evaluated and accepted measures for the risk assessment. The system of risk assessment was developed in the USA for the assessment of chemical substances and includes the identification of the hazard, the exposure assessment, the dose-response-assessment and the risk assessment. Basis for the risk assessment are detailed information of the chemical substance or the pathogen and epidemiologic data. Preventive strategies and risk management like the stop of production have great social and economic consequences. Risk management must be highly effective. In a historic review it is described that in the past of the former centuries infectious diseases were tried to prevent or control in a wrong manner in consequence of the lack of knowledge about the etiology of infectious diseases. Since Robert Koch there was the beginning of an effective risk assessment and management of infectious diseases. Long time in this century it was thought that infectious diseases wouldn't play any role for the public health. Now it is clear that infectious diseases remain the leading cause of death and disability in the developed and underdeveloped countries and that the risk potential is increasing. Therefore risk-assessment and risk-management developed formerly for chemical substances are necessary for infectious diseases. Specific criteria for-the risk assessment for infectious diseases which are different to chemical substances are discussed like illness related, pathogen related, host related, transmission related factors and general aspects. The most important measures for risk management are discussed. PMID- 9409919 TI - [The psychology of individual prevention of health risks]. AB - The object of the investigation is to diagnose the prototypical avoidance- and processing-mechanisms of risk factors for health and their psychological basis. The investigation is based on an explorative pilot study (n = 30 persons) and a psychological representative study (n = 415 persons; 208 men, 207 women; age: 28 60 years). PMID- 9409918 TI - [Toxicologic risk assessment and prevention: rational and irrational approaches]. AB - Worldwide, the management and evaluation of risks caused by chemical compounds are handled by the aid of threshold concentrations below of which one can be sure that no biological effect whatsoever can be observed. At the working place, we use the maximally tolerated concentration of the chemicals (MAK) and, in addition, the biologically tolerated concentrations (BTC) of the compounds either in blood or in other body fluids to which a male and/or female worker is exposed. These threshold concentrations should cover any toxic effect including on the one hand mere deviations of clinical chemical values without a disturbed function, i.e. symptoms of a disease as well as, on the other hand, carcinogenic, mutagenic and even allergic effects. Threshold concentrations, however, exist only for acute and chronic toxic effects and not for carcinogenic and/or mutagenic effects. In these cases, again, worldwide, the concept of minimizing the risks by the exposure is preferred since no toxicologist can be found to assure a "safe" concentration of a chemical compound that exert carcinogenic and/or mutagenic effects. With respect to these effects a proven carcinogenic and/or mutagenic effect in human beings must be discerned from a suspected effect on the basis of animal experiments or in vitro models. However, there exist also paradigms of a clearcut connection between a chemical substance or its metabolites causing carcinogenic and/or mutagenic effects in model experiments from which a clearcut suspect of similar reactions after the exposure of human beings can be drawn. In general, carcinogenic risks are overestimated in our societies. Following the data of experienced British epidemiologist most tumor diseases can be traced back to food consumption, beverages and tobacco and even sexual behavior must be ranked as cause for tumors before the rare exposure to dangerous chemicals at the working place. It is worthwhile to mention that natural toxins produced by bacteria and even infectious diseases or diseases caused by parasites are far more serious than the exposure to any man made chemical product including the Seveso poison, i.e. 2,3,7,8-TCDD, and related compounds. Vice-versa, the assumption that naturally occurring poisons could be neglected may lead to fatal experiences as for instances the outbreak of St-Anton's fire, i.e. the gangraeneous type of ergot alacaloide intoxication after having swallowed claviceps purpurea poisoned "Musli" produced by rye collected in the fields and ground in a hand mill. In Middle-Europe, since 1880, when the threshold of 0.1% claviceps purpurea in rye was established, no poisonous epidemia of St. Anton's fire was observed. PMID- 9409920 TI - [Education for responsible health behavior]. AB - 34 nursery school teachers, 40 primary school teachers, and 40 parents of primary school children were interviewed. Based on the results of these interviews, standardized questionnaires were developed and answered by 100 mothers of primary school children. Main results are as follows. Most of the nursery school teachers, primary school teachers, and mothers don't show great interest in health-related information, despite the fact that they lack knowledge in this area. Health risks which children could be educated to know, like unhealthy nutrition, are underestimated, whereas ecological risks which cannot be affected are overrated. Nursery and primary school teachers hold that parents are responsible for the health education of their children. Consequently, health related behaviour in kindergarten and health-related teaching in school are unsatisfactory. Mothers, however, expect schools to teach health-related topics concerning dependencies of all sorts--nicotine, alcohol, illegal drugs, and medication. Nursery school teachers as well as mothers demonstrate little systematic control and few sanctions when it comes to teaching children hygienic behaviour. Therefore, most children show satisfactory preventive behaviour only with regard to brushing their teeth. There are clear deficits in other areas. The mothers act only partly as good role models--for example, many mothers smoke when their children are around. Additionally, mothers don't regularly talk to their children about health-related topics. PMID- 9409921 TI - [Risk of exposure of children to indoor air allergens]. AB - The development of allergic sensitization and disease in children depends mainly upon the genetic predisposition, the time and extent of exposure against allergens and upon environmental factors. Allergen avoidance in infancy substantially lowers the frequency of sensitization and allergic manifestations. In Germany mite allergens are the most important indoor allergens followed by cat allergens and fungal allergens. Private houses bear the greatest risk of exposure to mite allergens. Nevertheless in day nurseries there is a considerable amount of dust samples with mite allergen levels greater than 10 micrograms/g dust (18% of mattresses, 15% of soft toys, pillows etc.). Cat allergen levels in day nurseries and schools are similar to those in houses where a cat has never been kept. There is a correlation between the percentage of children who have cats at home and the cat allergen levels in dust. In an own study the mean numbers of fungal particles in dust samples from day nurseries were higher compared with those from private houses. Higher numbers of fungal particles concerned mostly day nurseries with flat roof and moisture problems. We conclude that generally private houses offer the highest risk of exposure to indoor allergens. In addition public places like day nurseries or schools must be taken into consideration as significant allergen sources. PMID- 9409922 TI - [Procedures for hand hygiene in German-speaking countries]. AB - According to the field of application, strategies for the prevention of the transfer of microbial skin flora from the hands must consider the various categories of flora: transient, resident or stemming from infected lesions on the hands (infection flora). Depending on the species and virulence of the microorganism and of the susceptibility of the infection target, transient flora may or may not be of pathogenic importance. In contrast, resident skin flora is usually regarded as pathogenic only under certain circumstances such as in surgery, especially with transplantation of foreign bodies and in highly susceptible hosts. Microorganisms stemming from infected lesions are of proven pathogenicity. In the non-surgical field, only the transient and infection flora from the hands play a role. Such lesions are an absolute contraindication for patient-care, preparation of pharmaceuticals or foodstuff. In some procedures, the transmission of transient flora can be prevented by use of the non-touch technique ("instruments instead of fingers") or by the intelligent use of protective gloves. Hands already contaminated may be rendered safe by procedures for the elimination of transients such as handwashing, hygienic handwash and hygienic hand rub (in the order of increasing efficacy). Among all useable chemicals, ethanol, isopropanol and n-propanol (in the order of increasing efficacy) are the strongest and fastest agents. Furthermore, the duration of treatment (between 30 and 60 s) significantly influences the achievable reduction of microbial release. According to the new European standards (CEN) for testing chemical disinfectants and antiseptics, products for hygienic handwash must be significantly more efficacious than unmedicated soap, on artificially contaminated hands. In contrast, products for the hygienic hand rub must not be significantly less efficacious than a reference disinfection including isopropanol 60% vol rubbed onto the hands of the same volunteers during 1 min. By this, the average reduction of microbial release amounts to 4.2 to 4.4 lg, in our hands. The effectiveness of procedures for the hygienic handwash is usually significantly lower than that of alcoholic rubs. Therefore, in hospitals, they can be used only in certain indications such as patient care in reverse isolation, preparation of pharmaceuticals or foodstuff. In the surgical field, where not only transient but also resident flora is a cause of post-operative infection, the microbial release from the hands of the surgical team into the surgical wound must be prevented by using surgical gloves. Because of frequent glove lesions, a surgical hand disinfection is usually performed before donning gloves to keep a possible inoculumn as small as possible. Also in this field of application, alcoholic rubs proved to be significantly more effective than washing hands with antiseptic detergents. There exists a strong positive correlation of the reduction of microbial release and the duration of hand treatment, between 1 and 5 min. The European test standards (CEN) require products for surgical hand disinfection to be at least as efficacious as a reference disinfection of clean hands, which are constantly rubbed and kept wet with n-propanol 60% vol during 3 min. By this, the achievable average reduction of the microbial release ranges between 2.0 and 2.4 lg. In contrast, antiseptic washing procedures with preparations containing povidone-iodine, chlorhexidine gluconate or triclosan reduce the bacterial release within 2-5 min only by 0.5 to 1.2 lg. Some of them exert a bacteriostatic sustaining effect which is not found with alcoholic preparations. This, however, is not necessary with the latter as the initial bacterial reduction is that strong that restitution of the skin flora takes > 3 hours. Alcoholic preparations are at least as tolerable for the skin as antiseptic detergents, if not better, if they contain suitable emollients. Because dilution renders alcohols i PMID- 9409923 TI - [Hand hygiene in Latin countries]. PMID- 9409924 TI - [Possibilities and limits of modern rapid procedures for process control of cleaning and disinfection methods]. AB - In food production and processing, hygienic measures and their control gain more and more importance. The disadvantage of the commonly used control methods is their time-consuming process providing results after 24 hours at the earliest. From all microbiological rapid methods developed in the past, the bioluminescence method is a suitable hygienic measure for detection of adenosine triphosphate (ATP). In this test, ATP from microorganisms and ATP from somatic cells is detected. Within 2 minutes cleaned and disinfected surfaces can be reliably evaluated. Different equipment was tested in the laboratory and in a meat processing plant. The quality of the hygiene evaluation of the cleaned surfaces is decisively depending on the surface structure of the materials and on the product residues remaining in the peak-to-valley heights. Therefore, limiting values for ATP shall be determined individually for each plant. In this case, the bioluminescence method can be recommended as suitable tool for hygiene monitoring. PMID- 9409925 TI - [Public health--development and changes in the concept]. AB - Being a term of technical language originally, nowadays hygiene is widespread in everyday language. The linguistic law governing such transitions has been well described in philology. Consequently the change of a term from technical to standard language is characterized by the loss of its denotation, its exact and specific meaning, while its connotation is increasing. Initially hygiene was used in the sense of, maintenance and improvement of health', relating to the private as well as public and working area. As all kinds of dirt and pollution have been recognized as a risk to health, hygiene more and more became a synonym of cleanliness and sterility. This sense of word has soon been widened by, free of something that does not belong'. Therefore a metaphorical use of the term became possible, the meaning of which may differ completely from the technical sense. It is the question, in what way the conception of the field gets influenced by these facts of the matter. PMID- 9409926 TI - Perspectives in developmental biology. 2nd biennial meeting of the Sociedad Mexicana de Biologia del Desarrollo. Mexico City, June 12-14, 1995. Abstracts. PMID- 9409927 TI - [Quality of housing in relation with human health. I: Building characteristics of the houses in the Marchesian region]. AB - In view of the fact that a considerable proportion of the population particularly at risk (the aged, children, the sick, etc.) spend many hours of the day indoors, the authors report the results of an investigation carried out on the building characteristics of 1833 houses sampled in four towns in the Marches region. The questionnaire of Fusillo et al. was used. The results show an excellent situation with regard both to the quality of outdoor air and acoustic pollution (only 27% of the sample complained of the vicinity of their house to zones of intense traffic). The low index of overcrowding (0.7 inhabitants per room) as well as the overall surface area of the houses (in 60% more than 100 m2), the large number of bathrooms (almost 50% of the houses with two, and another 15% with three or more), and the prevalence of detached houses are factors that show how the Marchean population pay particular attention to building criteria. However, even though from the structural standpoint the housing situation might seem a very good one, specific problems can be found, also common to other Italian regions, regarding building techniques and the materials adopted (e.g. the use of reinforced concrete, which together with the undoubted advantages it offers also has a number of disadvantages). The authors therefore stress the need for incentivating a bioecologically based building tradition, even if this is economically more costly, and give guidelines on how to conciliate the bioecological and economic aspects with regard to choice and use of materials. Finally, they express the hope that the "EC" symbol foreseen for building materials by Law No. 246 of 21 April 1993 will be adequately used in the ways laid down in appendix A to that law. PMID- 9409929 TI - [Presence of zinc in food and diet in pediatric age]. AB - The authors determined the zinc content in the foods of eight diets recommended for different groups of subjects between four and a half months and twelve years. Then the average daily intake was calculated at any age. The highest values were found in meat products, the lowest ones in fruit and vegetables. The average levels of zinc intake were almost always upper than L.A.R.N. in both male and female. PMID- 9409928 TI - [Impregnation of uniforms with permethrin as a mean of protection of working personnel exposed to contact with hematophagous arthropods]. AB - Laboratory bioassays were conducted to evaluate the effectiveness of permethrin impregnated clothing against biting arthropods. Military battle dress uniforms were impregnated at a rate of 0.12-0.15 mg active ingredient (A.I.)/cm2. Gas chromatographic analysis of cloth patches treated showed that over 50% of the chemical remained after 6 cycles of washing. This amount of A.I. (0.06-0.07 mg/cm2) was effective in providing protection against Aedes aegypti and Ixodes ricinus. Mosquitoes and ticks were exposed respectively for 30 sec-3 min and 5-10 min to permethrin-impregnated uniforms: although the knockdown effect diminished with washing, the remaining permethrin provided 100% mortality of the mosquitoes and over 90% mortality of the ticks. PMID- 9409930 TI - [Economy and management: what doctors think. A national survey]. AB - INTRODUCTION: As managerial issues are a crucial component of the recent health care reform (Dec Leg.vo 502/92), it is urgent to collect information on physicians' knowledge, attitudes and interests on management and economics. Aim of the national survey presented here is to make a contribution in this area. METHODS: A survey based on 18 items questionnaires with closed-ended questions sent to 1,720 physicians (general practitioners, neurologists and psychiatrists). RESULTS: Response rate was 42%. Only 13% of respondents stated that the Reform will not affect physicians' work. More than 50% of our sample thought that economic and management skills may contribute to the improvement of the NHS. Younger physicians and senior consultants are more aware of the role of management in their profession, while GPs tend to be sceptical. DISCUSSION: Economic and management issues are no longer ignored by physicians. There is the opportunity to develop further training programs for physicians aimed at improving their managerial skills. PMID- 9409931 TI - [New aspects of the Italian health system and the management of medical units and facilities. 3: proposals for the training of personnel for the management of health units and facilities]. AB - In this delicate phase of transformation of the whole health organization and public employment in Italy several training aspects of managers deserve our attention. The target to reach is to achieve in all managers: a standard basic knowledge level in the various essential matters for a modern health structures management; a common language and a common strategic and logistic planning capabilities and solving methodologies of management problems. Plans for pluriprofessional, multidisciplinary, interactive short refresher course and for postgraduate training courses were presented. PMID- 9409932 TI - [The importance of epidemiology investigation]. PMID- 9409933 TI - [Arterial hypertension in children and adolescents]. PMID- 9409934 TI - [Blood pressure normal values in children and adolescents according to age and height]. AB - The assessment of blood pressure in children and adolescents is of great importance in order to gain a better understanding of its pattern of evolution. The authors publish the normal values of blood pressure in Portuguese children and adolescents, according to age and sex, as well as the mean values of the 90th percentile, which are very important to separate normal children from those with high blood pressure. In this study the fifth Korotkoof sound is used to define diastolic blood pressure in all ages. New blood pressure tables are also published, for children and adolescents, that now include the height percentile for age and blood pressure. These new charts have been developed to guide practising clinicians in antihypertensive drug therapy, when indicated. PMID- 9409935 TI - [Prevalence, knowledge, treatment and control of arterial hypertension in Oporto, Portugal]. AB - The aim of this study was to assess the prevalence, awareness, treatment and control of hypertension among subjects above the age of 39 years living in the urban area of Oporto, Portugal. One hundred and seventy seven individuals from the community were selected by random digit dialing. Each subject was asked about his/her personal history of hypertension, antihypertensive treatment and had his/her blood pressure measured. The prevalence of hypertension was 57.1%, defined by systolic blood pressure (SBP) > or = 140 mm Hg and/or diastolic blood pressure (DBP) > or = 90 mm Hg and/or administration of current the antihypertensive medication. If the values defining hypertension were SBP > or = 160 mm Hg, and DBP > or = 95 mm Hg the prevalence would be 37.9%. The overall prevalence of hypertension was higher in females, but a slightly higher non significant value was found in males in the fifth and sixth decades. Among hypertensives, 62.7% were aware of their condition, 56.7% were treated, 84.2% of hypertensives treated were controlled (SBP < 160 mm Hg and DBP < 95 mm Hg) and 44.7% were very well controlled (SBP < 140 mm Hg and DBP < 90 mm Hg). The question "Are you hypertensive?" had a sensitivity of 62.7%, a specificity of 83.6% and an accuracy of 75.7%. In the preliminary results of this study of an urban population with a high prevalence of hypertension, the awareness of hypertension is similar to that described in the United States of America twenty years ago, the percentage of hypertensives treated is similar to the American percentage fifteen years ago and the percentage of hypertensives treated and controlled is close to the current American percentage. PMID- 9409937 TI - [Prevalence of several cardiovascular risk factors in a population in the municipality of Viseu]. AB - STUDY OBJECTIVE: To assess the prevalence of the main changeable cardiovascular risk factors (hypertension, hypercholesterolemia and smoking) in the population of the Viseu municipality. MATERIAL AND METHODS: The population was obtained through a publicity campaign in local radios and on lighted placards in the town of Viseu. We chose a set of volunteers above 20 years of age, who answered a questionnaire about smoking habits, academic qualifications, profession and residence. After the inquiry, the total cholesterol and blood pressure were determined. We considered hypertension (HBP) values > or = 140/90 mmHg and hypercholesterolemia > 200 mg/dl. RESULTS: 1852 persons were inquired (3.2% of the population of the Viseu municipality) 1173 of which were females. According to the age groups, we verified that the ages between 50 and 69 years represented 47.9% of the total amount of volunteers. In what concerns smoking habits, we found a prevalence of 9.1% (15.9% in males and 5.2% in females). In the study of the prevalence of HBP we found a value of 38.5%, higher in males (42.8%) than in females (35.9%). The prevalence of hypercholesterolemia found was 34.9% (no significant differences between the sexes). CONCLUSION: In comparison with other studies carried out in the Portuguese population, a low prevalence of smoking habits, high hypertension and similar hypercholesterolemia were found. PMID- 9409936 TI - [Risk factors for myocardial infarct: a case-control study in Oporto, Portugal]. AB - A case-control study of coronary heart disease (CHD) was conducted in Oporto, Portugal. The cases series consisted of 100 consecutive patients with first time acute myocardial infarction who were admitted to the Coronary and Intermediate Care Units of a major teaching hospital. The community controls were 198 individuals without evidence of CHD by the Rose questionnaire and electrocardiography, selected by random digit dialing, with a participation rate of 70%. Data was collected by trained interviewers using a structured questionnaire and blood samples were obtained for selected laboratory data. The main analysis was made through unconditional logistic regression with calculations of odds ratios (OR). Age, OR: 1.5 (95% CI: 0.8-2.9), male gender, OR: 6.7 (3.6-12.3), family history of premature CHD, OR: 2.4 (1.4-4.3), diabetes, OR: 3.4 (1.6-7.4), antecedents of hypertension, OR:1.9 (1.1-3.1), history of high cholesterol levels, OR: 2.3 (1.4-3.9), high levels of physical activity, OR: 2.0 (0.9-4.1) and tobacco smoking, OR: 8.3 (3.8-18.5) were significant risk factors of acute myocardial infarction. After controlling for demographic variables and for the mutual confounding effects of the risk factors, the investigated factors that remained significantly associated with the risk of developing acute myocardial infarction were male gender, OR: 17.3 (4.8-62.3), family history of CHD, OR: 3.6 (1.4-9.6), diabetes, OR: 4.2 (1.0-18.1), high cholesterol levels OR: 2.7 (1.2-6.1) and smoking habits, OR: 7.7 (1.8-32.4). A negative association with high education levels was significant after controlling for all the variables, OR: 0.01 (0.01-0.5). PMID- 9409938 TI - [The impact of transesophageal echocardiography in infective endocarditis]. PMID- 9409940 TI - [The control of blood pressure in a hypertensive population]. PMID- 9409939 TI - [Thromboembolic risk in chronic cardiac insufficiency]. PMID- 9409941 TI - Peripheral T-cell non-responsiveness in individuals exposed to Plasmodium falciparum malaria. PMID- 9409942 TI - Future of the CDS. Community Dental Service. PMID- 9409944 TI - Dental nurses: valued members of the team? PMID- 9409943 TI - Mike Grace talks to Tony Kilcoyne. Interview with the editor. PMID- 9409945 TI - Dentists who drink and drive: do they have a right to work? PMID- 9409946 TI - [Clinical characteristics and therapeutic perspectives of boutonneuse fever. Assessment of a caseload of 39 patients]. AB - Rickettsia conorii is the etiologic agent of Boutonneuse fever, a rickettsiosis of the spotted fever group that is endemic in Southern Italy. Chloramphenicol or tetracyclines are still the treatment of choice for this disease, and recently quinolones have also been utilized with success. In 1994-95, 39 otherwise healthy patients were admitted to our unit for Boutonneuse fever. They were treated, in random order, with quinolones (10 subjects received ciprofloxacin, 500 mg/12 h per os; 8 subjects received intravenous pefloxacin, 200 mg/12 h), or tetracyclines (21 subjects received intravenous rolitetracycline, 275 mg/12 h). Outcome was favorable in all cases and no significant complications were observed. However, in a significant number of cases, increased blood concentrations of glutamic-oxalacetic (68.4%) and glutamic-pyruvic (60.5%) transaminases were found. Above normal blood creatinine values were observed in 29.7% of the cases, and urinanalysis disclosed blood in 35.9% and proteins in 56.4% of the cases. Both tetracyclines and quinolones were well tolerated and effective, with apyrexia achieved after 2.7 +/- 0.1 days (mean +/- SEM). All patients were discharged after an average of 7.1 +/- 0.4 days. Liver and kidney function derangements seem to occur to some extent in the acute phase of Boutonneuse fever. This finding might partially explain the increased mortality rate reported for subjects with simultaneous systemic or organ diseases or when the administration of an effective antibiotic is delayed. Together with chloramphenicol and tetracyclines, quinolones might be considered as first line antibiotics. PMID- 9409947 TI - [Correlation between skinfold thickness and ultrasonography in the study of subcutaneous adipose tissue in females]. AB - The aim of this study was to verify if there is a 1:2 correlation between subcutaneous adipose tissue thickness measured by ultrasonography and skinfold caliper and if this correlation varies in function of the degree of obesity, subcutaneous fat thickness, and the area evaluated. Forty women (age 27.9 +/- 11.7 years, body mass index 28.75 +/- 5.40 Kg/m2, waist to hip ratio 0.77 +/- 0.06) underwent ultrasonographic measurement of subcutaneous adipose tissue and skinfold caliper measurement at nine different sites (bicipital, tricipital, subscapular, suprailiac, epigastric umbilical, hypogastric, gluteal, and femoral). Data analysis confirmed a significant correlation between measurements made by ultrasonography and skinfold plicometry at all sites when the patients were not subdivided according to body mass index or skinfold thickness. When they were subdivided on the basis of body mass index, a significant correlation was found for subjects with a body mass index < 30; when the index was > 30, the correlation was observed at only the subscapular and suprailiac sites, and to a lesser degree at the tricipital and femoral sites. Moreover, a highly significant correlation was found only at sites at which ultrasonographic thickness was not > 20 mm (p < 0.001) with an r value decreasing progressively from 0.685 (thickness < 10 mm) to 0.248 (thickness > 40 mm). Given the great variability of this correlation, we suggest that ultrasonography is preferable to plicometry for the measurement of fat. PMID- 9409948 TI - [Angiogenesis and antitumor therapy]. AB - We have examined the most recent literature dealing with the relationship between tumor growth and neovascularization (tumoral neoangiogenesis). It is quite evident that antiangiogenic therapy against solid tumors has an important theoretical basis. Many antiangiogenic substances are presently under study, and some inhibitors of angiogenesis, such as TNP-740, platelet factor 4, linomide, tecogalan, etc., are currently being tested in phase 1 or 2 clinical trials. These studies have, however, underscored the possibility of numerous side effects due to the lack of specificity of these substances against endothelial cells. The most promising and rational use of antiangiogenic factors in anticancer therapy seems to be their association with traditional chemo- or radiotherapy. Gene therapy also offers considerable theoretical promise as demonstrated by the transfection of thrombospondin 1 complementary DNA into a human breast carcinoma cell line. PMID- 9409949 TI - [Pyomyositis in Italy: report of a clinical case]. AB - Pyomyositis is an infection of the striated muscle seen frequently in Africa but rarely in Western countries with a temperate climate. Over the last few years it has been observed with increasing frequency, especially in immunocompromised hosts. An unusual case of pyomyositis in a 65-year-old immunocompetent woman is described. The disease emerged during septicemia caused by Staphylococcus aureus. It was associated with pleuropneumonia and affected two different and opposite groups of muscles. Diabetes mellitus, a known predisposing factor, was diagnosed during the infection. The diagnosis of pyomyositis was based on microbiological cultures, computed tomography, and radio-labelled granulocyte scintigraphy. Follow-up until recovery was based on computed tomography. Surgical drainage of abscesses was avoided thanks to early diagnosis and specific antibiotic therapy. PMID- 9409950 TI - [Behcet's disease and angioedema induced by ACE-inhibitors: 2 unusual pathological pictures]. PMID- 9409951 TI - 8th World Congress of the International Society for Brain Electromagnetic Topography (ISBET) with the KEY Foundation Symposium: Brain Fields in Psychiatry. Zurich, Switzerland, March 6-8, 1997. Abstracts. PMID- 9409952 TI - Oncology update: activity of docetaxel in single-agent and combination chemotherapy regimens. Proceedings of a symposium. Denver, Colorado. PMID- 9409953 TI - Management of Pediatric Asthma. Symposium proceedings. London, England, November 18, 1996. PMID- 9409954 TI - Breast abscess due to Corynebacterium striatum. AB - Corynebacterium striatum, a normal constituent of the skin flora, is rarely pathogenic. Previous reports of infection are few, and are mainly confined to immunosuppressed patients or those with indwelling prosthetic devices. We report a case in which the organism caused a recurrent breast abscess in a woman with normal immune function. The only previous reports of Corynebacterium striatum mastitis have been in cows. PMID- 9409955 TI - Bridging the neurosciences. PMID- 9409956 TI - Proceedings of the Plant Polysaccharide Symposium. Nantes, France, 17-19 July 1996. PMID- 9409957 TI - A new millenium of nutrition research. A celebration of the 90th birthday of Dr Elsie Widdowson. 21 October 1996. PMID- 9409958 TI - [The genetic implications in glaucoma]. PMID- 9409959 TI - [Growth factors in proliferative diabetic retinopathy]. AB - This work presents the possible implications of the angiogenic growth factors and some cell mediators in the initiation and development of the neovascular proliferation in diabetic retinopathy. According to the physiopathologic theories stated above, that are implied in the generation of proliferative diabetic retinopathy, here are some therapeutic experiments based on the action of the angiogenic growth factors. PMID- 9409960 TI - [Normal-tension glaucoma. Its diagnosis and treatment]. AB - The paper attempt a NPG definition in the light of the most recently data from literature. Modern investigation techniques (CDI, LDF, laser scan angiography) are summarized to support the presently most accepted pathogenic theory--the vascular theory. There are also reviewed the main clinical conditions to do differential diagnosis and main methods of its treatment. PMID- 9409961 TI - [The progression of primary operated glaucoma]. AB - OBJECTIVE: Study of the causes responsible for glaucoma progression in patients operated for glaucoma. MATERIAL AND METHOD: A series of 185 patients operated for primary open angle glaucoma were followed up for a mean period of 3.2 years. 46 patients had evidence of glaucoma progression: insufficient filtration was recognized as the cause in 23 patients, while blockage of the filtering bleb was responsible for the rest of 23 cases. RESULTS: Progression of glaucoma was detected in 19.32% of patients with functional filtering bleb; 44.1% of patients with intraocular pressure between 16 and 21 mmHg did not show disease progression; 8 out of 23 patients needed a new trabeculectomy; internal obstruction of the bleb was demonstrated in 8 cases; 4 patients needed additional medical therapy; internal revision of the bleb (3 cases) and a new trabeculectomy gave the best results; external obstruction of the bleb was found in 15 patients; 7 had a diffuse fibrovascular proliferation and 8 had an encapsulated bleb. We used a new trabeculectomy for diffuse fibrovascular proliferation and a cystotomy or a cystectomy for encapsulated bleb. The results were good in almost all cases. CONCLUSIONS: Glaucoma progression appears even with postoperative intraocular pressures within statistical normality; identification of the causes responsible for glaucoma progression insures individualized treatment with constant good results. PMID- 9409962 TI - [Biochemical aspects in the pathogenesis of senile cataract]. AB - The content of sialic acids has been investigated in the lacrimal liquid in patients with senile cataract as well as in the intracapsular extracted crystalline lenses. The highest level of sialic acids was detected in the lacrimal liquid and in the extracted crystalline lenses of patients with mature senile cataract. In this patients a higher frequency of the incipient senile cataract at the other eyeball has been noted. PMID- 9409963 TI - [Syphilitic parenchymatous keratitis]. AB - It shows a clinical observation about luetic bilateral keratitis at a young man 17 years old. The high-positive VDRL, the presence of the AT antigen, the bilateral low hearing perception and the left low hearing transmission associated with minimal dental malformations suggest the luetic congenital etiology of the disease. The AT positive-test at the patient's mother (which VDRL is negative) shows a tardive luetic infection during the pregnancy, that has determined the apparition at the fetus of congenital lately lues. The specific antiluetic treatment with Benzylpenicillinum kalicum and Prednisonums the evolution of the disease is spectacularly good. Six weeks after the beginning of the treatment V.A. is 1 at the both eyes and the neoformation vessels are completely obturated. The paper insists on the medical and social implications of the ignored complications of late lues. PMID- 9409964 TI - [The results and complications of artificial lens implantation in the anterior chamber]. AB - The purpose of the study is to make a comparison in the evolution of AC-IOL per primam and per secundam. MATERIAL AND METHOD: We reviewed 50 cases operated in the University Clinic of Ophthalmology from Cluj-Napoca since 1995-1996, 35 managed with primary and 15 with secondary AC-IOLs. RESULTS: Early and late postoperative complications are frequently present at the primary AC-IOLs. Explanation was performed in 2 cases. The functional results were superior at secondary AC-IOLs. CONCLUSIONS: AC-IOL is the solution to be chosen in case of intraoperative complication (loss of vitreous). Secondary AC-IOL is to be preferred because the postoperative complications are fewer and the functional results are better. PMID- 9409965 TI - [Genetic predisposition in glaucoma]. AB - The comparative study of the genetic factors implication in low-tension and high tension glaucoma showed the presence of glaucoma familial cases in both form. In 3 cases of low-tension glaucoma the presence of other familial cases of high tension glaucoma was revealed. The existence of hereditary cases of high-tension glaucoma in families with low-tension glaucoma suggests the existence of some common factors, responsible for producing both affections. On the other hand, the systematic research of the existent glaucomatous deficits at the patients with glaucoma familial cases deserves greater attention. PMID- 9409966 TI - [The Gronblad-Strandberg syndrome associated with macular degeneration]. AB - It presents one case of Strandberg-Gronblad syndrome associated with macular degeneration in atrophic phase. There are presented reason by general, local and pathogenic exam. It is underlined the particular clinical aspect of the macular degeneration in these cases, the severity of complications and therapeutical inefficiency. PMID- 9409967 TI - [Marginal keratitis in Behcet's disease]. AB - One female patient in twenty-six years old, presents corneal marginal relapsed at the left eye, aphthae of inferior eyelid and aphthous stomatitis in episodically fits repeated at three-four weeks. Also she presents disorders of the intestinal transit and thrombosis of the right inferior limb, fragility of the vessels, thoracic neurodermatitis with presternal erythema; these phenomenon are improvement with cortico and immunomodulator therapy. On insist about present of the marginal, relapsed ulcus in Behcet disease. PMID- 9409968 TI - [Behcet's disease with a fatal evolution]. AB - Behcet disease in an inflammatory systemic disease, of unknown etiology. Usually it is defined by a triad; buccal ulcerations, genital ulcerations and uveitis. Treatment is based on steroids and in difficult cases cyclosporina or cytotoxic agents could be used. We present a case of Behcet disease in a young male. The first symptoms were retrobulbar optic neuropathy, followed by buccal and genital ulcerations. For beginning the treatment was based on steroids, but the disease was more and more severe, so that the left eye lost its function because of an inflammatory maculopathy and new symptoms arose. We started treatment with cyclosporina (5 mg/kg/day), but we had to stopped it 3 months latter because of renal function's failure. PMID- 9409969 TI - [Ultramicroscopic studies in retinoblastoma]. AB - Were examined three cases with undifferentiated retinoblastoma. Were effected cross sections on the samples, remarking tumoral cells with normal aspect interspersed with tumoral necrosis cells. Tumoral cells of the undifferentiated retinoblastoma has low size and a poor cytoplasmic with a big nucleus that occupies nearly the whole cell; the ratio nucleus/cytoplasmic is increased. The cells are uni- or binucleus and the nucleus is monstrous deep invagination of the cover nucleus. Cytoplasmic has poor organelles and mitochondria are swelling following a high degree of hypoxia. Also tumoral cells on remark nontumoral cells (melanocyte). PMID- 9409970 TI - [The Rieger syndrome. A clinical study. A study of 4 generations in one family with the Rieger syndrome]. AB - Rieger syndrome is a rare disease with autosomal dominant inheritance (4q25-4q27) characterised by the presence of ocular and extraocular abnormalities. We present a family with Rieger syndrome on four generations, pointing the ocular and extraocular abnormalities. The most frequent complication is secondary glaucoma (50-60% of all cases) and the efficient treatment consists in utilising the betablockers and/or filtering operations with the possibility of antimetabolites help. PMID- 9409971 TI - [Persistent hyperplastic primary posterior vitreous]. AB - The clinical study done on 19 cases with posterior primitive vitreous persistency and hyperplasia showed the predilection affectation of youth (average age: 11.2 year old), the frequent lateral localization (17 cases), microcornea (7 eyes), hypermetropia (average value + 3D). The visual acuity was between 1 and unregistered device values, being a function of the papillar ant retinal lesion extension. The ophthalmoscopical aspect was balanced, from a simple prepapillar veil to complex ophthalmological syndromes. Etiopathogenetically, the affection is considered to be a embryogenesis flaw, appeared in the development of the primary hyaloid-vitreous complex, described by an incomplete resorption and a hyperplasia of its elements. The problems of differential diagnosis and treatment are extensively presented. PMID- 9409972 TI - [The treatment of penetrating eye injuries]. AB - Data about eye traumatism at the Republican Ophthalmologic Center are presented. 791 patients have been investigated: 52.3% with penetrated wounds; in 19.3% cases we detected foreign intraocular bodies. Fibrinolysin and dexamethazone have been introduced during operation in the case of fibrinoplastic inflammatory process with crystalline lens masses in the anterior camera. As a result we noted a positive evolution in the postoperative period. PMID- 9409973 TI - [Vitreous surgery in ocular traumatology]. AB - This study points out vitreous surgery's indications, technical means and results in various ocular traumatisms. Over one-year period, in the Charles Nicole Hospital in Rouen, vitreous surgery has been performed in 67 patients with ocular traumatisms. The surgical techniques, the intra- and postoperative complications are described and correlated to the preoperative ocular status. Results are significantly better in the group with iatrogenic traumatism than in the one with accidental traumatisms. Vitreo-retinal surgery is the only therapeutical method in various ocular traumatisms intraretinal foreign bodies, nonmagnetic intraocular foreign bodies, posttraumatic tractional retinal detachment, giant retinal tears, posttraumatic intravitreal hemorrhage without spontaneous resolution. Without vitreo-retinal surgery, visual function's loss would have occurred. PMID- 9409974 TI - [Postoperative flat anterior chamber. The therapeutic approach]. AB - OBJECTIVE: The study of clinical entities followed by postoperative flat anterior chamber and their therapeutical possibilities. Retrospective study of all flat anterior chamber cases in a series of 315 patients operated for glaucoma. There were 16 flat anterior chamber cases with ocular hypotony, 3 cases with pupillary block and 9 cases with ciliary block. Stage I flat anterior chamber cases with ocular hypotony responded well to medical treatment. The most efficient surgical procedures were the application of additional sutures and excision of the over filtering bleb. The most frequent complications are: secondary cataract (75%), filtering failure (25%) and uveitis (18.75%). Flat anterior chamber cases with pupillary block resulted from obstruction and were managed with a new peripheral iridectomy. Flat anterior chamber cases with ciliary block had a severe evolution and didn't show any improvement under medical therapy. All cases were managed successfully by vitreous surgery (especially using pars plana aspiration). The incidence of postoperative flat anterior chamber can be reduced by accurate surgical procedure. Most of flat anterior chamber cases with ocular hypotony had a favorable evolution under medical treatment. Cases associated with intraocular hypertension needed surgical management. PMID- 9409975 TI - [Trabeculectomy with scleral drainage and diathermy coagulation: a new therapeutic variant]. AB - Several failures with postoperative reappearance of a hypertonia after trabeculectomy, determined the authors to bring some improvement. For avoidance the scarring, on practice diathermal clotting of one middle channel of 1.2 mm, using a spatula. Were obtained well outcomes with normal ocular pressure in twenty-three cases. PMID- 9409976 TI - [The correction of myopic astigmatism by refractive keratotomy]. AB - PURPOSE: To determine the efficacy and the predictability of refractive keratotomy in the myopic astigmatism. METHOD: 15 patients (23 eyes) who had been operated on with myopic astigmatism; 4 eyes with simple astigmatism and 19 eyes with compound astigmatism, 1 patient had myopia at one eye and myopic astigmatism at the other. It were performed tangential incisions ("T"), curved incisions ("C") and simple Ruiz with or without radial incisions. The patients were reviewed after 3.6 and 12 month from the operation. RESULTS: Better visual acuity with no correction. The only postoperative complication was under correction. CONCLUSIONS: Refractive keratotomy is recommended in myopic astigmatism and especially with anisometropia. PMID- 9409977 TI - [Update in ophthalmology (Prague, 10-13 March 1997)]. PMID- 9409978 TI - [The clinical value of electromyography of the external ocular muscles]. AB - The paper shows the electromyographic contribution looking the physiopathological study of the extrinsic ocular muscles. The electromyogram outcome and its repetition allows to establish one topographic diagnosis of the extrinsic ocular muscles disorders: proper ocular external muscle disorders (ocular myopathia). neuromuscular transmission disorders (myasthenia); lesions of peripheric neurons (neurogenic paralysis); lesions of central innervation (supranuclear lesions). PMID- 9409979 TI - [Functional recovery in strabismus]. AB - The paper enumerates the objectives of functional recovery in strabismus and the approaches to the achievement of these objectives. The goal in the treatment of the strabismus is obtaining a binocular sight with the remake of the divergence convergence synergism. A three stage scheme for the treatment is made: 1st stage- the treatment of the amblyopia; 2nd stage--the correction of the strabismus deviation; 3rd stage--the recovery of the binocular sight. The conclusion is that the healing of the strabismus with "restitutio ad integrum" is a distant desire. PMID- 9409980 TI - [The pathogenesis of cotton-wool spots]. AB - Cotton-wool spots have been recognized as distinct ophthalmoscopic entities, yet the pathogenesis of these lesions has not been fully appreciated until now. Cotton-wool spots should be regarded as localised accumulations of axoplasmic debris in the retinal nerve-fibre lay, which result from interruption of axoplasmic transport. Cotton-wool spots could be found especially in diabetic retinopathy, retinopathy from nephropathy, collagen diseases, retinal vein occlusions and cachexia. Any factor responsible for local interruption of axonal flow in the retinal nerve-fibre layer will give rise to similar accumulations, although, pathogenically speaking, the cause is usually ischaemia. PMID- 9409981 TI - [Clinical correlations between the incidence of diabetic retinopathy and diabetic nephropathy]. AB - In this study we included 651 patients with recently diagnosed diabetes mellitus registered at the Centre for Diabetes Care from Timisoara in 1995. All the patients were from Timisoara and the county of Timis. They had insulin-dependent and noninsulin-dependent diabetes mellitus. We studied the eye lesions ophthalmoscopically, retinophotographically and angiographically and we correlated these informations with the lesions of diabetic nephropathy. We also made an evaluation of the functional implications of the ocular lesions. PMID- 9409982 TI - [Conservative treatment in intraocular tumor in the remaining eye]. AB - We present here, the case of a monophtalm patient, with the diagnostic of an intraocular tumour, situation when we have chosen a combination of conservative therapeutically means. Enucleation, as a radical method of treatment, is out of the question. We discuss, by turns, the main alternatives of conservative treatment: sclerochoroido-retinectomy, temporary fixation on the supraiacent sclera of radioactive discs, photocoagulation and cryocoagulation as single methods of treatment, justifying the present option: laser treatment associated with cryocoagulation and immunotherapy. We review some basic concepts of immunology, insisting upon the systemic and topical administration of IL-2. Up to the present, IL-2 is administrated topically only in pulmonary cancer and in cerebral tumours, after the operation, on the excision zones and in cutaneous malignant melanoma, with very good results. In ophthalmology, this is the first therapeutically attempt, using IL-2 in a case of intraocular tumour. The postoperative status and the results on short term put an end to this paper. PMID- 9409983 TI - [The recovery of visual performance in strabismus with eccentric fixation]. AB - The purpose of this work is to realise a general view about the diagnosis, therapeutic methods and the results obtained in eccentric fixation strabismus. We realised a six years retrospective study on 144 eccentric strabismus, watching dynamic the evolution by testing: the orthophoric status, visual acuity, binocular vision and the type of fixation. Medium follow un time was of 19.6 months. In the end of the treatment we notice: visual acuity between 0.1-0.5 in 14% cases; central fixation in 52 children (36%); first degree of binocular vision in 15 children (10.4%) and second degree in 5 cases (3.47%). This study showed that the early installation of eccentric fixation strabismus was in 53% cases before reaching one year old and in 90% before two years old. The functional results was negative influenced by the delay in presentation (42% come after 2 years) and the difficulties in family cooperation (46.5% were followed up lesenban 6 months). PMID- 9409984 TI - [Ocular myopathy. A study of 4 cases]. AB - Four cases of ocular myopathy are presented. Their clinical, electromyographical and hystological signs are discussed comparatively with clinical forms described in the literature. One case was of pure ocular myopathy, without the affectation of other skeletal muscles. The other three cases were included in the descendent ocular myopathy, presenting a motor deficiency at the centure and inferior member levels. In all cases modifyings have been found in the EMG trajectory and the muscular biopsy of skeletal and/or ocular muscles. PMID- 9409985 TI - [Familial congenital aniridia]. AB - We have studied a family that has been presenting cases of congenital aniridia for three generations. A specific trace found in this family is the varied expression of the pathological gene as some members showed bilateral congenital aniridia, others showed unilateral aniridia with coloboma or just bilateral coloboma aspect. Aniridia can be found either as a "de novo" mutation in a family or as a continuous transmission between generations reproducing the dominant autosomal model. The ocular manifestations of this anomaly include: defects of the cornea, glaucoma, lens subluxations, hypoplasia of the fovea, nystagmus. PMID- 9409986 TI - [The role of zinc in the appearance of cataract]. AB - The level of zinc in 259 crystalline lenses removed after extraction of cataract (senile and traumatic) has been determined. The lowest concentration of zinc in crystalline lenses has been detected in patients with mature senile cataract while the highest concentration-in patients with traumatic cataract. Since many enzymes contain zinc we can suppose that their activity decrease as senile cataract develops. Higher level of zinc in traumatic cataract indicates moderate disturbances of crystalline lens metabolism as compared with senile cataract. PMID- 9409987 TI - [Adenocarcinoma of Moll's glands]. AB - A Moll gland adenocarcinoma case is presented. The female patient was 47 years old and the affection was localized at the inferior eyelid. The histopathological exam shows the presence of intense eosinophil cells with gland-shape disposition. The histopathological exam was necessary both for the diagnosis and for the establishment of the degree of differentiation of the tumour, emphasizing the extension of infiltration localization for the neighbouring structures. PMID- 9409988 TI - [Anatomicopathological studies in retinoblastoma]. AB - For a fifteen-years period it has been examined fourteen cases with retinoblastoma, twelve of them have been undifferentiated forms and two cases differentiated forms in rosette, in undifferentiated retinoblastoma has been observed low size cells with poor cytoplasm and big nucleus that occupied near the whole cell; the nucleus to cytoplasm ratio is increased. The cellular density is irregular and most important perivascular. The tumoral parenchyma is condensed in perivascular areas, the tumoral stroma is poor, amorphous or mucinous. The necrosis areas are frequently but very import out in undifferentiated retinoblastoma. The tumoral vascularity is increased. In some cases occur melanin pigment that belong to the pigment epithelium without the existence of tumoral proliferation. PMID- 9409989 TI - [Trauma from light induced by the operating microscope in the rat retina]. AB - The study proposes to analyze the lesions show to the retina level of the rat exposes to light of an operate microscope Zeiss Op Mi 220 (without filters). The conditions of the experiment was created such as the photochemical effect of the radiation to predominate more than thermic effect. Rat's retinae was visualized through photonic and electronic microscopy. The images show that the maximal lesions are situated to the level of the retinal pigment epithelium and of photoreceptors cells. The histological aspects presume the action of the free radicals in the production of the lesions. PMID- 9409990 TI - [Current concepts in the treatment of recent and late posttraumatic glaucoma]. AB - The ocular hypertonia is a frequent complication of ocular trauma. It may be precocious, if it appears immediately after or within a few days since the trauma, or it may appear slowly in the following weeks or months. The choose of the therapy is made after the physiopathogenic mechanisms. The urge of the surgical intervention is difficult to be established. In all cases the supervision of the ocular tonus in time is recommended. PMID- 9409991 TI - [The posterior chamber implant per secundam]. AB - The purpose of the study is to show the efficacy and advantages secondary implant of PC-IOL in certain conditions. MATERIAL AND METHOD: We studied 8 aphakic eyes which had been operated on in the past for senile and traumatic cataract and who desired an secondary implantation of IOL. In 3 causes it was performed Yag laser capsulotomy for secondary cataract. The functional results in all cases were between 0.7-1 with optimal correction. CONCLUSION: Secondary implantation of PC IOL is a good solution in aphakic eyes, in certain ocular conditions. PMID- 9409992 TI - [Retrobulbar optic neuropathy in multiple sclerosis in remission only with parenteral treatment]. AB - The authors are presenting a case diagnosed with retrobulbar neuritis in the left eye secondary to multiple sclerosis. The symptoms submitted only under infusions with corticosteroids. Despite the lack of methylprednisolone, hydrocortisone was used successfully. This case supports the efficiency of intravenous treatment of optic neuritis as modern therapy in neuro-ophthalmology. PMID- 9409993 TI - [A model follow-up record for diabetic retinopathy (DR). Its staging in the order of severity]. AB - Based on up-to-data in the literature, a practical possibility to classify as possible the opportunity, in order to appreciate as fast, correct and efficient as possible the opportunity for sending patients to laser therapy. The defining elements for every important sign of diabetic retinopathy are analyzed. They are grouped on their evolutive stages (according to the international standards) in the order their gravity and urge in applying laser therapy. Then a personal model for a follow-up sheet (algorithmic type) is presented. Ist main advantage consists of the fact that the recognized and only mentioned signs lead to the stage grouping, to the evaluation of therapeutical possibilities and may replace a photo-document still hard to be obtained. PMID- 9409994 TI - [Guidelines for standardized diagnosis, therapy and after-care of thyroid cancer]. PMID- 9409995 TI - [Minimally invasive treatment of abscesses by CT-controlled drainage with a basket catheter system]. AB - PURPOSE: To examine whether the percutaneous drainage of abscess formations by a new basket catheter system is usefull. MATERIAL AND METHODS: 58 patients with abscesses in different locations and origins have been treated by an interventional radiologic procedure. On the whole 77 basket-catheters were placed under CT-guidance into abscess formations of different size and localization. 36 patients developed an abscess after surgery, two patients achieved abscess drainage after embolisation of a tumor. In 20 patients the abscess was a complication of a septic infectious disease. RESULTS: Open surgery was avoided in 41/58 patients of these patients 9/41 received only percutaneous drainage and 32/41 patients suffering from illness were given antibiotic medication according to the resistogram in combination with percutaneous drainage. 17/58 patients required secondary surgery but with a lower risk because of the smaller abscess volume and the better clinical constitution after percutaneous drainage. CONCLUSIONS: A minimally invasive management of abscesses using a basket-catheter system is successful even in localisations deep inside the body and hard to reach. An irreversible catheter occlusion followed by the implantation of a new catheter system could be avoided in all patients. PMID- 9409996 TI - [Radiologic appearance of primary anomalies of the lymphatic vessel system of the lung. Review and personal results]. AB - Congenital disorders of the lymphatics of the lung are rare. On the basis of a literature review and our own experiences the disorders are divided into four groups and their radiographic findings are described. Pulmonary lymphangiectasia shows not typical signs. As an expression of the underlying pathophysiologic processes CT shows thickening of interlobular septs, interstitial edema and pleural effusions. Neither architectural distraction nor thickening of intralobular septs was seen. Lymphangioleiomyomatosis shows rather typical findings with multiple thin-walled bullae. In contrast to the opinion that architectural distraction is not seen in LAM we sometimes found signs of fibrosis. Disseminated pulmonary lymphangioma is characterised by proliferation of lymph vessels. CT-findings of this disorder have not been described before. CT shows multiple disseminated nodules of up to 2.5 cm accompanied by multiple bullae of similar distribution and size. PMID- 9409997 TI - [Hepatic pseudolesion caused by puncture]. AB - We report the diagnosis and differential diagnosis of a hepatic pseudolesion due to percutaneous liver biopsy in a 30-year-old female patient with known chronic hepatitis C and renal insufficiency. In the course of transplant preparation, an abdominal spiral-CT examination pre and post i.v.-contrast injection as well as an angiography with CT-hepaticography and CT-portography were performed. In these examinations a 1 cm, hepatocellular-carcinoma mimicking liver lesion was found; it was hypodense in the CT-portography and showed a marked enhancement in the CT hepaticography. This "pseudolesion", which was supposed to be due to the liver biopsy, resolved spontaneously. PMID- 9409998 TI - [Preoperative hook wire marking of nonpalpable suspected breast lesions]. AB - 45 clinical occult breast lesions in 43 patients were preoperatively localised using a mammographic unit, a hole compression plate, and a hookwire canula. 20 carcinomas were found. The positive predictive value of the mammography was 44%, a good result. Serious complications were absend and only one lesion was missed. The preoperative method is effective, rapid, and safe for detecting and localising occult breast lesions. The number operative explorations may be reduced by close contact to the radiologic colleagues and more primary stereotactic biopsies. PMID- 9409999 TI - [Mammography after breast preserving therapy and reconstruction. Experiences in routine practice]. AB - In the last few years we have seen, in daily routine, many different methods of operation and their complications. We want to show several diagnostical difficulties. Between January 1995 and July 1996 we evaluated all our patients retrospectively. They were treated by several methods of operation. The women were examined with the routine possibilities such as mammography, sonography, fine needle biopsy, and clinical examination. We examined 161 patients in the last 18 months. They were treated by breast-conserving therapy (127 women) and augmentation (34 women) at the age of 29-82 (O 55 years). We listed complications as well as early and late recurrences. The time after operation was 1/2-22 years. In our long-term experience we found fewer recurrences after breast-conserving treatment when the women also received radiation too (only operation 16%, operation and radiation 7.6%). For the future it is hoped to eliminate local recurrences by new operation methods (augmentation plasty with flaps, TEM flaps). Two patients with local recurrence are still alive after 22 years, so that not only local diagnosis and state of nodes are important also the individual immune reaction. PMID- 9410000 TI - [Development of a CT data-based score for prediction of fracture risk in osteoporosis]. AB - It should be examined how far BMD semiquantitative (profile analysis) and qualitative datas (architecture of spongy bone, age of patient) can be combined in one score in order to improve the assessment of fracture risk. SE-QCT was performed in 220 patients with a mean age of 55.8 (33-84) years from whom conventional X-ray images of the thoracic and lumbar spine were available. In the axial scans spongiosa architecture was classified and a density profile analysis was carried out. This was followed by gradation of BMD values, different types of spongiosa architecture, profile analysis and age of patient to a numerical score. This was compared to the number of fractures, whereby the patients were separated into three groups: group I = no fracture, group II = one fracture, group III = more than one fracture. The BMD values, types of spongiosa architecture, semiquantitative profile analysis can be significantly assigned to the groups I and II (p < 0.02), groups I and III (p < 0.001), and the groups II and III (p < 0.05). By combining BMD values, architecture of spongy bone, density profile analysis, and age of patients without fracture, scale 8-12 = patients with or without fracture, scale 13-16 = patients with at least one fracture [corrected]. PMID- 9410001 TI - [Preoperative radiotherapy for prevention of heterotopic ossifications after hip endoprosthesis]. AB - Preliminary results of a prospective study which investigates the efficacy of preoperative radiotherapy (RT) for prevention of heterotopic ossification (HO) after total hip arthroplasty are summarized. A total number of 20 hip joints (18 patients) were irradiated with a single dose of 6.0 Gy < 4 hours before the surgical procedure. After an average observation period of 6.9 months (range: 6 11 months) the results were evaluated using radiological and clinical criteria. At follow-up, one case (5%) had evidence of high-grade HO (> Brooker grade II). The functional outcome quantified with the Harris score was improved by an average of 37.9 points. The authors conclude that preoperative RT is an effective alternative for postoperative irradiation. PMID- 9410002 TI - [Percutaneous radiotherapy of bone metastases]. AB - Bone metastases causing pain syndromes and imminent pathologic fractures are among the main reasons for radiotherapy in patients suffering from malignant tumors. The indication is much influenced by the radiologic findings. Surgical methods are to be chosen in the first place in cases of pathologic fractures or patients with a high risk of such fractures. Different authors recommend various therapeutic regimens. Effective pain control can be achieved with one single dose of radiation. Doses of 6 to 40 Gy applied in one to 19 days are also efficient. Side effects, especially nausea and vomiting occur in 25% of cases; this number rises to 50% in cases of half body irradiation. Visible changes of bone mineral density may be noticed about 6 weeks after termination of radiotherapy. About 70% of osteolytic metastases show progressive sclerosis whereas osteosclerotic lesions may show both increase or decline of bone mass. In spite many years of experience the optimal strategy for radiation therapy of bone metastasis has not been defined; further studies are needed. PMID- 9410003 TI - [Unexpected computerized tomography findings in neoplasm staging of a patient with plasmacytoma]. AB - A case of a 69-year-old male patient is described. Staging CT brought unexpected findings with therapeutic consequences. Pneumoperitoneum was the most important sign. A pneumoretroperitoneum and a dilatation of the colon sigmoldeum were also found. Intraoperatively a perforated sigmadiverticulitis was detected. Therefore, one should pay attention to extraskeletal changes during the staging of multiple myeloma. PMID- 9410005 TI - [Comparison of film-screen combinations with contrast detail diagram and interactive image analysis. 2: Linear assessment of grey scale ranges with interactive image analysis]. AB - The following three screen-film combinations were compared: a) a combination of anticrossover film and UV-light emitting screens, b) a combination of blue-light emitting screens and film, and c) a conventional green fluorescing screen-film combination. Radiographs of a specially designed plexiglass phantom (0.2 x 0.2 x 0.12 m3) with bar patterns of lead and plaster and of air, respectively were obtained using the following parameters: 12 pulse generator, 0.6 mm focus size, 4.7 mm aluminum pre-filter, a grid with 40 lines/cm (12:1) and a focus-detector distance of 1.15 m. Image analysis was performed using an IBAS system and a Zeiss Kontron computer. Display conditions were the following: display distance 0.12 m, a vario film objective 35/70 (Zeiss), a video camera tube with a PbO photocathode, 625 lines (Siemens Heimann), an IBAS image matrix of 512 x 512 pixels with a resolution of 7 lines/mm, the projected matrix area was 5000 microns2. Grey scale ranges were measured on a line perpendicular to the grouped bar patterns. The difference between the maximum and minimum density value served as signal. The spatial resolution of the detector system was measured when the signal value was three times higher than the standard deviation of the means of multiple density measurements. The results showed considerable advantages of the two new screen-film combinations as compared to the conventional screen-film combination. The result was contradictory to the findings with pure visual assessment of thresholds (part I) that had found no differences. The authors concluded that (automatic) interactive image analysis algorithms serve as an objective measure and are specifically advantageous when small differences in image quality are to be evaluated. PMID- 9410004 TI - [Diagnostic angiography in life-threatening colonic hemorrhage in Crohn disease]. AB - Life-threatening colonic hemorrhage is a very rare condition in Crohn's disease. Lower gastrointestinal hemorrhage in Crohn's disease occurs in 0.6-2.5% of cases. In a patient presenting with severe gastrointestinal bleeding after ileocolectomy, mesenteric angiography provided precise localisation of the bleeding site in the area of the splenic flexure. PMID- 9410006 TI - [Speed S sensitivity of film-screen systems and mode of action of different automatic illumination in general practice. I: Film-screen systems]. AB - A new test procedure is presented to check the correspondence between various screen-film systems (SFS) with the specification of the manufacturer in respect to the speed S according to DIN 6867-1 or rather which dose K is required for the SFS in dependence on the X-ray tube voltage (kV) to give the net density Dn = 1.00, it is based on measurements of dose and density using on X-ray equipment with water as an absorber and scatter test tool. The characteristic curves of the automatic exposure control (AEC) can be recorded and compared with those of SFS, i.e., how is the dose K in the image-receptor-plane regulated by the AEC in dependence on different radiation qualities. It is also possible to check the modes of operation of SFS and AEC with sufficient accuracy but lower time and energy requirements. The consequences of finding the right values to test the X ray equipment under DIN 6868-50/-3 and for constancy-testing according to DIN 6868-3 are discussed. PMID- 9410007 TI - [Assignment of a component of serine proteinase fluorescence spectrum to a cluster of tryptophan residues]. AB - The two log-normal spectral components in, alpha-chymotrypsin, trypsinogen and beta-trypsin fluorescence and the single component of chymotrypsinogen A emission were analyzed using characteristics of microenvironment of atoms of indolic fluorophores of individual tryptophan residues in the three-dimensional structures of the proteins. Tryptophan residues (eight ones in chymotrypsinogen A and alpha-chymotrypsin and four ones in trypsinogen and beta-trypsin) in these homologous proteins form three clusters with resonance excitation energy exchange between fluorophores within each of them. In all the proteins, Trp51 and 141 determine the shorter-wavelength spectral components (ca. 330 nm) and Trp215 and 237 emit as longer-wavelength ones (ca. 340 nm). Excited states of fluorophores of Trp27, 29, 207 (cluster A) and 172 (in cluster B) are deactivated by the near disulfide bonds. The longer-wavelength component is quenched in chymotrypsinogen emission due probably to the formation of an electron trap including Asn91 amide and several water molecules. The contribution of longer-wavelength component in trypsin spectrum increases comparing with that of trypsinogen due to some little changes in localization and a huge rise of mobility of Met180 sulfur atom nearly Trp215 in trypsin. PMID- 9410008 TI - [Theoretical conformational analysis of cardioactive peptides. I. Tetrapeptide Phe1-Met2-Arg3-Phe4-NH2 and pentapeptide Leu1-Pro2-Leu3-Arg4- NH2]. AB - By means of semi-empirical method, an a priori conformational analysis of a cardioactive peptides was carried out. Calculations yielded the values of all dihedral angles of the backbone and side chains of the peptides various forms as well as intra- and interresidue interaction energies. PMID- 9410009 TI - [Theoretical conformational analysis of cardioactive peptides. II. Nonapeptide Met1-Asn2-Tyr3-Leu4-Ala5-Phe6-Pro7-Arg8-Met9-NH 2]. AB - Theoretical conformational analysis was carried out for the cardioactive nonapeptide Met1-Met9-NH2. Calculations yielded the values of all dihedral angles of the back bone and side chains of the nonapeptide various forms as well as intra- and interresidue interaction energies. PMID- 9410010 TI - [Effect of chain length on formation of a double-layer structure of Z-(Gly-Pro Gly)n-OMe oligotripeptides]. AB - Effect of chain length on the formation of the double-layered sheet in synthetic oligotripeptides Z-(Gly-Pro-Gly)n-OMe (n = 1,2,3,4) in the solid state has been studied by the IR-spectroscopy method. The major spectral features characteristic for the polymer structure was found in the oligomer spectra beginning with that of hexapeptide (n = 2). So a minimum peptide length when peptide chains are still able to adopt the double-layered sheet conformation in the solid state includes two triplets. The result is similar to that we have revealed early by investigation of the collagen-like triple helix formation in the oligomer series Z-(Gly-Pro-Pro)n-OMe. The double-layered sheet structure of the hexapeptide was found completely regular as that of the polymer. So the formation of the double layered sheet structure unlike the formation of the triple-helical structure occurs at once without gradual ordering of interpeptide bonds and molecular packing by further increasing of the triplet number in oligomer chains. Additional information concerning specific incorporation of bound water in the double-layered sheet and water influence on peptide packing has been derived from analyses of hydration effect on the oligotripeptide IR-spectra especially on the components of the amide I band. PMID- 9410011 TI - [Dependence of the rate of EPR spin-lattice relaxation of certain proteins on temperatures in the interval 6-22 K]. AB - It was shown, that rate of spin-lattice relaxation EPR signals of ferredoxin in temperature interval 9-22 K is proportional to T5. The mechanism of spin-lattice relaxation of non crystalline systems that explains the experimentally observed strange peculiarities of temperature dependence of relaxation rate, for a number of globular proteins, has been considered. PMID- 9410012 TI - [Theory of Mossbauer effect in hydrated globular proteins]. AB - The spectrum of Mossbauer absorption by the label incorporated into macromolecule is constructed. The dependence of the spectrum parameters on the hydration degree is found. The results are compared with experiments. The fluid flow inside the globule, excited by rotational vibrations of the globule surface is described by the linearized Debye-Brinkman equation. The local coefficients of rotational and translational diffusion of the globule fragments are found. The hydration coefficient is determined by the ratio of the effective inertia of the porous sphere with the fluid to the inertia of solid sphere. PMID- 9410013 TI - [Formation and melting of ordered structures in denatured myoglobin with differing water content: differential scanning calorimetry method]. AB - The possibility of superstructure formation in denatured globular proteins has been studied through the temperature dependence of the absolute values of heat capacity in the myoglobin-water system with water content from 80 to 25% in the temperature range 20-160 degrees C. For all the composition range studied it is found that after the denaturation of myoglobin the new regular structures with reversible melting are formed. These structures are similar in properties to the thermotropic gels in concentrated myoglobin solutions. Decrease of the water content influences the formation of these non-native ordered structures in denatured protein and increases the dispersion of its formation and melting temperatures. PMID- 9410014 TI - [Heat-induced transitions in proteins]. AB - The heat-induced relaxation of globule conformational was studied under mild physiological conditions by extra-high frequency dielectrometry in the range of dispersion of free water. It is shown that macromolecules possess a functionally significant molecular memory, which is due to their spatial organization and, probably, to water associates. PMID- 9410015 TI - [Dynamic superhelicity in vivo and in vitro]. AB - As it was shown in studies of J. Dubochet using the recently developed method of three-dimensional cryo-microscopy, the interaction of RNA polymerase with plasmid gives rise to a superhelical structure. In this process, two RNA polymerase molecules are located at opposite ends of the plasmid at the tops of the superhelix. The dynamic features of this process are considered, which may be only one of many similar processes, in which energy dissipation during the directed movement of DNA is reduced to a minimum. It is assumed that, upon screwing of two double helices into each other, a three- dimensional mechanical transmission is formed, owing to the spatial features of the double helices comprising the superhelix, which further facilitates this twisting process. PMID- 9410016 TI - [Interconnection of hierarchical compactization levels as the basis for functional organization of chromatin]. AB - Recent data on chromatin structural organization are presented. In the light of present concept of chromatin structure the problem of hierarchical levels of DNA packaging within chromatin is discussed. Particular emphasis has been placed on the correlation between the changes occurring at different DNA compactization levels within topological domains and chromatin functional activity. PMID- 9410017 TI - [Modeling changes in membrane physical changes during binding of a substrate to membrane proteins]. AB - Change in the physical properties of the bilayer membranes due to binding of the hormone molecules by the membrane proteins are investigated. They are manifested substantially only for definite protein concentration and are determined by the dimensions of the zones of the distorted structure which are located near free receptor molecules as well as the receptor-substratum complex. The mechanisms of membrane and blosensors sensitivity to hormones are discussed. PMID- 9410018 TI - [Effect of ribonuclease and lysozyme on physical properties of the lipid bilayer]. AB - The effect of ribonuclease and lysozyme on pyrene excimerization in liposomes composed of phosphatidylcholine and diphosphatidylglycerol has been studied. Microviscosity of bulk and annular lipids was found to increase upon the formation of lipid-protein complexes. PMID- 9410020 TI - [Effect of heliogeophysical factors on the biological activity of Staphylococcus aureus]. AB - In 1988-89, an experimental was carried out to study the effect of heliogeophysical factors on the biological activity of Staphylococcus aureus, one of the most widespread causative agents of infectious diseases in man and animals. For comparison, both individual heliogeophysical factors and interrelated phenomena in the system Sun-Earth arising from solar flashes were used. Two types of solar flashes were revealed. A near-annual cycle of changes in DNase activity of staphylococci in vitro was revealed, which correlates with the cycle of changes in electron concentration of layer F2 of the ionosphere. The correlation coefficient is 0,96%. It was found that the threshold of susceptibility of test-microorganisms to heliogeophysical influences is different in different years. There is an "amplitude window" of the influence whose upper boundary varies in different periods. PMID- 9410019 TI - [Differences in osmoregulation of resistant and nonresistant fibroblasts in hypotonic media and effect of verapamil on osmoregulation]. AB - The osmoregulation of resistant and nonresistant striped hairyfooted hamster fibroblasts in hypotonic medium was studied. It was found that the fibroblasts of a resistant population completely regulate their volume 1 min after diluting the cell suspension with water. Verapamil, an inhibitor of Pgp channels, caused equal swelling of both fibroblast populations in isotonic medium. It was shown that the water entering the cells together with verapamil increases equally the volume of both cell types and remains bound to both resistant and nonresistant fibroblasts after the termination of cell volume regulation. PMID- 9410021 TI - [Photosensitizing and photoprotective properties of extracts from groups of medicinal plants]. AB - Investigation of photosensitization and photoprotection induced by plant extracts was carried out. A group of plants affected human central nervous system was studied in detail. Efficiency of plants as photoprotectors and photosensitizers was tested in the frame of the influence of their extracts on the yield of photochemiluminescence of Gly-Trp solutions. Photosensitization was studied under irradiation with light lambda > 280 nm and lambda > 320 nm, as well as with monochromatic light lambda=313, 365, 405 and 436 nm. All of the plants studied acted as photoprotectors in low concentration and as photosensitizers in high concentration. The efficiency of photoprotection and photosensitization was evaluated with regard to single dose of plant extracts and their concentration in human organism. The effect decreases in the following consequence of plants: Leonurus > Hypericum > Aralia > Schizandra > Echinopanax > Eleutherococcus > Valeriana > Panax ginseng. Photosensitization is due to the components of plant extracts which have strong absorbtion at the high wavelength range. The mechanism of photosensitization was suggested. Singlet oxygen generated by photoexcited compounds is the main species resulted in chemiluminescence. Superoxide radicals does not contribute significantly to the chemiluminescence formation. PMID- 9410022 TI - [Electrical reverberator on the excited surface of the myocardium]. AB - The modelling of spiral autowave (reverberator) on excited surface of the myocardium which appear in the fibrillation of heart was studied. The shape of reverberator is found. The kinematic parameters of spiral autowave were given in our paper. PMID- 9410023 TI - [Antitumor effect of a weak superlow frequency stochastic magnetic field with 1/f spectrum]. AB - Antiblastomic effect of low strength low frequency fluctuating magnetic field with 1/f power spectrum (1/f MF) analogous to spectra of normal biological fluctuations is studied experimentally. For the first time it is demonstrated that stabilization of cell-cell surface interactions as well as suppression of mitotic activity of malignant cells exposed to 1/f MF may result in considerable inhibition of cancer growth in animals. These effects were shown to be absent in weak low frequency magnetic fields with spectra other than 1/f. PMID- 9410024 TI - [Differential reassociation curves in studying genome structure]. AB - Reassociation kinetics and molecular structure of hepatocytes DNA of rabbits Shinshilla, Marder breeds and their hybrids have been measured. It is shown that hybrids genome is not the simple sum of donors genome. The hybrids genome has new specific feature of molecular organisation which different from donors genome. PMID- 9410025 TI - [Synthesis of oligopeptides from the polar amino acids in water media by the combined action of weak electric and magnetic fields]. AB - The effect of weak electromagnetic fields on aqueos solutions of polar amino acids leads to the formation of stable products of the amino acid condensation reaction. These products have a definite amino acid composition and retention time in high-performance liquid chromatography. PMID- 9410026 TI - [Effect of molecular dynamics on the photoinduced electron transfer in eosin myoglobin complex]. AB - The temperature dependence of the rate constant of photoinduced electron transfer in the modified eosin-myoglobin complex by monitoring of the phosphorescence quenching of eosin is measured. The values of electron transfer rate constants are equal 10(2) + 10(3) s-1 in the temperature region 150-200 K. The kinetics of relaxation of the maximum of the time-resolved phosphorescence spectra of eosin on apomyoglobin is measured in the same temperature range. The solvation relaxation of the time-resolved phosphorescence spectra is nonexponential. The characteristic times of the solvation relaxation are given 10(-2) + 10(-4) s-1, that correlate with the time of electron transfer in this system. It was observed the "acceleration" of the relaxation rate of the time-resolved phosphorescence spectra of eosin in metmyoglobin due to nonequilibrium photoinduced electron transfer. The role of the matrix dynamics in photoinduced electron transfer in proteins is discussed. PMID- 9410027 TI - [NMR study of beta-structures in the compact non-native states of globular proteins]. AB - By the high resolution NMR method was made comparative research of carboanhydrase B, alpha-lactalbumin and apomyoglobin in a molten globular state. It is shown that though in an observed spectra the typical signals at 5.7-5 ppm, responding to the signals of alpha-CH protons of the beta-structure, method of spin diffusion proves that after all is taking place in carboanhydrase B and alpha lactalbumin, that corresponds with the data of CD. PMID- 9410028 TI - [Stabilization of albumin by CaCl2- and MgCl2-regulated interaction of macromolecules: a study by the spin label method]. AB - Effective thermodynamic activation parameters of spin-label transitions between microsurroundings in water-protein matrix of modified molecules of serum albumin are determined in 0.01 M acetate buffer, pH 5.6, in CaCl2 and MgCl2 concentration range from 10(-3) to 5 M and protein concentration 90-110 mg/ml. The "melting" of water-protein matrix being accompanied by disjoining pressure in water-protein matrix and native protein structure stability alterations is registered in the salt concentration range 0.1-0.3 M. The phenomena revealed are interpreted as phase transition from solution of protein clusters at the low salt concentrations to the "salting out" form of protein solution at high salt concentrations or to the solution in which interactions between proteins are stabilized by repulsive structural forces of residual hydration shells of the bound cations. This transition realizes through the intermediate (critical type) state characterized by weak protein-protein interactions. The flexibility of water-protein matrix and its permeability for the salt ions and water molecules is more pronounced in this region of moderate (approximately equal to 0.3 M) CaCl2 and MgCl2 concentrations. PMID- 9410029 TI - [Structural changes in serum albumin induced by SHF heating]. AB - A comparative study of the effects of heating by microwave radiation and water bath heating up to temperature 35-75 degrees C on the structural state of bovine serum albumin is presented. Microwave radiation perturbs the surface of an albumin molecule. This essentially affects aggregation properties of bovine serum albumin and enhances structural transformations which accompany the process of thermal denaturation. PMID- 9410030 TI - [Effect of the volumetric dielectric properties of the medium on stability of the globular proteins and membranes]. AB - Analysis of the temperature dependences of the denaturation enthalpy of three domains of albumin molecule showed significant distinctions in the destabilizing effect of ethanol on certain domains of the nondefatted and defatted albumin molecules. A linear relationships were found among the parameters of proteins and membranes stability and bulk dielectric constants of the solvents. It has been suggested that the dielectric characteristics of the medium play a significant role in the maintaining of the certain conformation of proteins and membranes. PMID- 9410031 TI - [A study of the effect of the adsorbed molecules of concanavalin A on characteristics of the bound water in the hydrate coat of the disperse silica by 1H NMR of the frozen water suspensions]. AB - The experimental technique is presented which allows the investigation of water at the adsorbent-biopolymer interface by record of temperature dependencies of 1H NMR signals during the heating of previously frozen water suspensions. It was found that the adsorption of protein conconovalin A resulted in twofold decrease of strong bound water amount and in 70% increase of loosely bound one. The mean value of Gibbs energy decreased twice for strong bound water and increased twice for loosely bound one. The external layer of hydrate shell of silica particles with adsorbed conconovalin A was organized by protein molecules. This conclusion is based on the observed total increase of loosely bound water amount, as a result, of protein adsorption, although its amount with respect to a protein molecule decreased. PMID- 9410032 TI - [Information capacity of the nucleotide sequences and their fragments]. AB - The problem of determining the information content of nucleotide sequences is discussed. Exact expressions for the reconstitution of higher-order frequency dictionaries from lower-order once were obtained by the maximum entropy method. In form, they are analogous to superpositional approximations known in statistical physics. The features of entropy characteristics of real nucleotide sequences are described that reliably distinguish them from random texts. Methods for comparing the information content of frequency dictionaries and assessing the residual uncertainty of the text at the known frequency dictionary are proposed. PMID- 9410033 TI - [Integral equations of the theory of liquid in the study of hydration of macromolecules]. AB - An efficient numerical algorithm for solving integral equations of the theory of liquid in the RISM approximation for infinitely diluted solution of macromolecules with a large number of atoms is proposed. The algorithm is based on applying the non-stationary interactive methods for solving systems of linear algebraic equations. Using this technique we have calculated the solvent-solute atom-atom correlation functions for short fragment of DNA duplex of varying length in aqueous solution. PMID- 9410034 TI - [Hydration of a fragment of the B-form of DNA duplex: d(GGGGG). A method of integral equations of the theory of liquid]. AB - Based on the numerical algorithm proposed by us in [1] for solving integral equations of the theory of liquids in the RISM approximation we calculate all of the solvent-solute atom-atom correlation functions for a fragment of the DNA duplex d(GGGGG) in infinitely diluted aqueous solution. The obtained results are compared with available experimental data and results from computer simulations. PMID- 9410036 TI - [Mathematical model of bilirubin transport]. AB - The mathematical model of the bilirubin transport through liver is suggested. The model is sufficiently reflects the peculiarities of the bilirubin behavior under the different physiological conditions. The model is described by the system of difference-differential equations. The coefficients of the intensity of bilirubin transfer in normal liver are calculated. PMID- 9410035 TI - [Effect of metal cations on the copper induced peroxidation of the low density lipoproteins]. AB - The effect of metal cations on copper-catalyzed lipid peroxidation (LPO) of low density lipoproteins (LDL) was examined. The presence of metal cations in the incubation media containing LDL (0.8 mg protein/ml) and CuSO4 (0-80 microM) influenced on LPO of LDL as evident by the measurement of TBARS. With the concentrations of CuSO4 less than 10 microM, the metal cations caused an increase in LDL peroxidation. Zn2+ appeared to be the most effective inductor, Mn2+ was less effective, and the influence of Ca2+ and Mg2+ was insignificant. With greater CuSO4 concentrations Mg2+ showed no effect on TBARS formation in LDL while the addition of other nontransition metal cations to the incubation mixture led to the inhibition of LDL peroxidation. The capacity for inhibition decreased in the row Mn2+ > Zn2+ > Ca2+ > Mg2+. The possible mechanism explaining these results may be in the competition of metal ions for copper binding sites on LDL. Our results allow to suggest the existence of two types of copper binding sites on LDL, tight-binding sites which are non-effective in LPO and effective weak binding sites. PMID- 9410037 TI - [Effect of structural analogs of platelet activating factor on the intracellular signal transduction in murine peritoneal neutrophils and macrophages of P388D1 line]. AB - Direct and modulate effects of platelet activating factor (PAF), its structural analogues and ATP on primary and second processes at peritoneal neutrophils and P388D1 cells activation has been studied. The effect of compounds was evaluated on changes in Ca2+ transport and generation of reactive oxygen species. It was shown, that the synthetic analogues of MS series interact with PAF receptor, mobilize Ca2+ from thapsigargin-dependent intracellular stores and inhibit Ca2+ response on PAF in both types of cells. Unlike PAF the analogues do not induce the formations of reactive oxygen species in neutrophils and inhibit the PMA induced respiratory burst. The activation of pyrinoreceptor of P388D1 cells by exogenous ATP does not inhibit PAF induced Ca2+ rise in cytoplasm, though partly releases Ca2+ from the same store. PMID- 9410038 TI - [Acid-induced hemolysis of human erythrocytes]. AB - The acid-induced haemolysis of human erythrocytes has been studied in the isoosmotic media (0.14 M NaCl) at pH 2.0-3.5. The apparent rate constant of this process increased and the time of lysis of half of the cells decreased after spectrin denaturation in the presence of ethanol. The time of the haemolysis diminished in the presence of human serum albumin, which interacted with the cell surface, and after erythrocyte modification by glutaric aldehyde. On the contrary, the hyperosmotic contraction of erythrocytes in the NaCl, KCl, or sucrose containing medium stabilized the cells. The activation energy of the process of acid-induced haemolysis determined by the temperature dependence of the haemolysis time was 69 +/- 4 kJ/mol. The cell volume increasing which induced by proton increasing, occurred at higher pH values (pH 5.5-5.0) as compared with pH inducing cell lysis. We suggest that not only osmotic mechanism takes part in the acid-induced haemolysis. PMID- 9410039 TI - [A cross-bridge model for the artificial mobile systems]. AB - A simple kinetic scheme of the mechano-chemical cycle of the cross-bridge is proposed. It was found on the experiments with using of caged ATP and the experiments with the limited number of the myosin molecules in motility assay in vitro. The model reproduces the results of the experiments in motility assay in vitro and allows to investigate the actin-myosin complexes at an external load condition. PMID- 9410040 TI - [Liberation of additional heat during muscle contraction (Fenn effect)]. AB - The calculation of additional heat evolved during contraction of muscle is presented. It is supposed that the cause of arise of additional heat is the transformation in heat of the potential energy which accumulates in actin filament in the process of its torsion. PMID- 9410041 TI - [Modulated extremely high frequency electromagnetic radiation of low intensity activates or inhibits respiratory burst in neutrophils depending on modulation frequency]. AB - The influence of low-intensity modulated electromagnetic radiation of extremely high frequencies (EHF EMR) on synergistic reaction of calcium ionophore A23187 and phorbol ester PMA in activation of the respiratory burst of the peritoneal neutrophils of mice line NMRI was investigated. The production of reactive oxygen species by the neutrophils was estimated by luminol-dependent chemiluminescence technique. The cells were irradiated in the far field zone of the channel radiator for 20 min in the presence of A23187 and then were activated by PMA after switching off the irradiation. It was shown, that continuous EHF EMR (50 microW/cm2) inhibited quasi-resonantly the synergistic reaction. The maximum effect was about 25% at carrier frequency of 41.95 GHz. Modulated radiation with carrier frequency of 41.95 GHz and modulation frequency of 1 Hz activated the synergistic reaction, but at modulation frequencies of 0.1, 16 and 50 Hz inhibited one. At fixed modulation frequency of 1 Hz the nonlinear dependence of the effect on the carrier frequency was found. The synergistic reaction was activated in the frequency range of 41.95-42.05 GHz and was inhibited at the frequencies of 41.8-41.9 GHz. The effect was observed only at raised intracellular free calcium concentration and at calcium fluxes through plasma membrane. The obtained results prove the possibility of control over cell functioning by low-intensity modulated EHF EMR, presumably, manipulating by connected systems of enzyme reactions. PMID- 9410042 TI - [A model of spherical quasi-epicardium for mapping of the electrical potential of the heart]. AB - Possibility of determination of the heart electric potential maps on the spherical quasiepicardium from the body surface potentials measured by means of multiple leads, is estimated. The simplified models of ventricular depolarization electric generator considered in this study, consist of sets of uniform double layers representing basic configurations of the true electric generator of the heart, while allowing an analytical calculation of the multipole components, as well as potential of the generator. The contribution of the generator multipole components to the potential on the spherical quasiepicardium is evaluated. The results of the study show that it is reasonable to use the multipole components up to the 3rd order for noninvasive mapping of the potential on the spherical quasiepicardium with the radius which provides tangential touching of the quasiepicardium with the precordium surface. Because of bringing the potential observation region nearer to the generator and improving concentricity of this region with respect to the generator, the quasiepicardium mapping technique allows revealing much more complicated generator configurations than those found out by straightforward estimation of the potential measured on the chest surface, thereby facilitating more accurate noninvasive recognition of the heart electrophysiological state. PMID- 9410043 TI - [A model of the biological oscillator as the basis for the coordination of motor activity]. AB - System mechanisms, that underlied motor activity coordinations were investigated on the model of biological oscillator. It has been shown how the value of phase resettings induced by single stimuli sequences depends on the stimulation location in the cycle. The time of reaction to the stimulus has been determined as the function of stimulus location. In addition, the character of transitional process, after which the parameters of initial rhythmic activity completely become normal, has been determined. The data obtained have been interpreted for the model of temporal motor control, which was suggested by A. Wing and A. Kristofferson. PMID- 9410044 TI - [Biophysical aspects of emergence and development of pathological conditions]. AB - The molecular mechanisms of rise, development, and correction of some uninfectious illness of ungenetical nature and conception of ligand pathology are analyzed. The influence of molecular geometry change upon the pathology development and biophysical aspects of drugs searching are considered. PMID- 9410045 TI - [Anti-meningococcal polysaccharide vaccines]. PMID- 9410046 TI - [Etiology of enteritis in a university general hospital in Barcelona (1992 1995)]. AB - BACKGROUND: The aim of the study was to describe the etiology of enteropathogenic agents over a four-year period (1992-1995) in a University Hospital in Barcelona. METHODS: We studied 12,793 stool samples, 4519 were obtained from patients under 15 years and 8274 were obtained from patients over 14 years. The specimens were examined for bacteriological, parasitological and virological enteropathogens. RESULTS: In 3380 specimens of 12,793 stool samples studied were identified an enteropathogen (26.4%). Polymicrobial associations were observed in the 6.8% of the cases. Pathogens were identified in 45% of children samples and 16.3% of adults samples. The etiological enteritis agents more frequently detected in the paediatric patients were Campylobacter (13.5%), rotavirus (11.3%) and Salmonella (10.2%); and Salmonella (4.9%), Campylobacter (3.1%) and Giardia intestinalis (2.1%) in adults. Cryptosporidium (13.5%) was the most frequent cause of gastrointestinal tract infections in HIV-infected subjects. In the children with stools positives, the presence of red and white blood cells were more frequent than the adults with stools positives (73% versus 26.6%). CONCLUSIONS: The enteropathogenic agents such as Campylobacter, Salmonella, and Giardia were the most frequent cause of gastroenteritis in our environment. In the children, rotavirus infections predominated during the cold months. The most frequent cause of gastroenteritis in HIV-infected patients was Cryptosporidium followed by Campylobacter. PMID- 9410047 TI - [Double-blind clinical trial comparing 5 days of dirithromycin versus 7 days of diacetylmidecamycin in acute bronchitis and acute exacerbations of chronic bronchitis]. AB - BACKGROUND: Dirythromycin has several pharmacokinetic characteristics (long half life and high tissue concentrations) which suggest the possibility of administering shorter treatments than those conventionally used. The aim of this study was to determine and compare the efficacy of a 5 day treatment with dirythromycin once a day, versus diacetylmidecamycin twice a day over 7 days in the treatment of patients with acute bronchitis and acute exacerbations of chronic bronchitis. METHODS: A parallel, multicentric, randomized, double blind clinical study was carried out in 8 Spanish hospitals. RESULTS: One hundred seventy-four patients were included in the study, with 87 (80 evaluable) being randomly assigned to receive dirythromycin (500 mg/day over 5 days) and 87 (83 evaluable) diacetylmidecamycin (600 mg, twice daily over 7 days). A favorable symptomatic response (cure or improvement) was observed in 72/80 of the first group (90%) and in 74/83 (89.2%) of the second group. No statistically significant differences were found in the efficacy and safety between the two treatment groups in either the evaluable patients or on intention to treat analysis. CONCLUSIONS: The results of this study suggest that the administration of dirythromycin, once a day over 5 days, is as safe and effective as diacetylmidecamycin, twice a day over 7 days, in the treatment of acute bronchitis and acute exacerbations of chronic bronchitis. PMID- 9410048 TI - [Isolation of cytomegalovirus from peripheral blood by shell-vial culture. Technical considerations]. AB - BACKGROUND: We performed an study of the possible benefits derived from the use of two vials, incubated for 24 and 72 h respectively, in the isolation of CMV from the leukocyte polymorphonuclear (LPMN) of peripheral blood (viremia) in immunosuppressed patients, using the shell-vial culture technique. MATERIAL AND METHODS: The blood samples with EDTA were fractionated by sedimentation with saline dextran. The LPMN-rich upper layer was used for the isolation of CMV in the shell-vial cell culture (MRC-5 cell line) and to prepare the antigenemia pp65 assay. The cultures were stained at 18-24 h (first vial) and 72 h (second vial) with an indirect immunofluorescence assay with a monoclonal antibody against p72 CMV antigen. RESULTS: We studied 878 blood samples of which 247 (28.1%) were positive for CMV. Of the positive samples, 211 (85.4%) were detected in the first vial, and 220 (89%) in the second vial. Neither of the two vials detected all positive samples. An overall toxicity of 3%, in the positive samples, and 8.5% in positive samples was detected, respectively. Of the 28 toxic samples, 25.9% were detected in the first vial and 85.1% in the second (p < 0.001). The use of the second vial increased the number of positive samples in 14.5% (36 blood samples). CONCLUSIONS: In view of the results obtained in this study it would seem that, in order to obtain the maximum diagnostic yield in the isolation of CMV from LPMN of peripheral blood in immunosuppressed patients by means of the shell-vial culture, it is necessary to inoculate all the LPMN extracted into two vials. The lengthening of the incubation period up to 72 h for one of the vials leads to an increased in the isolation of CMV of 14.5% with a low overall toxicity. PMID- 9410049 TI - [Prevalence of E. coli O157:H7 in a pediatric population in Bogota, D.C. with acute gastroenteritis]. AB - BACKGROUND: The aim of this study was to determine the prevalence of E. coli O157:H7 in pediatric patients with acute gastroenteritis (AGE). PATIENTS AND METHODS: A prospective epidemiologic study was performed in cases and controls to detect the germ in fecal samples of 300 children with AGE in Bogota, D.C. The stools of the patients and 85 controls were seeded in MacConkey-sorbitol agar. The sorbitol negative colonies were biotyped and seroagglutinated with a latex test for E. coli O157:H7. Antimicrobial susceptibility was performed after using the Bauer & Kirby method. RESULTS: Of the 300 diarrhea specimens analyzed, 14 strains corresponded to E. coli O157:H7 with a prevalence rate of 4.7% in children with AGE. The prevalence was 1.14%, the excess of risk of presenting E. coli O157:H7 was 14% in children with AGE. One child developed SUH. In three of the 85 controls E. coli O157:H7 was isolated, with a prevalence rate of 3.53%. The mean age of the 14 patients was 21 months (range: 3 months to 7 years) and the mean length of diarrhea in the children was 2 days. All the isolations of E. coli O157:H7 were sorbitol negative and indol negative with only 2 strains being MUG negative. DISCUSSION: This epidemiologic study has, for the first time, demonstrated the strong prevalence of E. coli O157:H7 in Colombia. The clinical data do not show a common pattern. Whether the strains isolated in the study are verotoxin producers remains to be demonstrated. PMID- 9410050 TI - [Meningococcal infection caused by Neisseria meningitidis serogroup C]. AB - BACKGROUND: In recent years an increase has been observed in the prevalence of meningococcal infection by Neisseria meningitidis serogroup C and in the appearance of strains with moderate resistance to penicillin. PATIENTS AND METHODS: A microbiologic study of the cases of meningococcal infection of serogroup C treated from 1995 to 1996 in the health care area of Ferrol (La Coruna, Spain) was carried out. RESULTS AND CONCLUSIONS: Twenty-nine cases were detected in 1995 and 28 in 1996. Meningococcal infection was observed in patients ranging from 8 months to 21 years of age (mean 5.7 years). Distribution by sex was homogeneous. Two patients died. According to the clinical presentation, 11 were sepsis (38%), 4 meningitis (14%) and 14 both processes (48%). In 4 LCR samples, the analytical study was normal with posterior positive culture results. The detection of bacterial antigen by latex agglutination in CSF only detected 32% of the cases. MIC study determined that 11 strains (38%) presented moderate resistance to penicillin, 9 with a MIC of 0.12 microgram/ml, one with a MIC of 0.25 microgram/ml and another with a MIC of 0.5 microgram/ml. In all the cases the strains were sensitive to cefotaxime (MIC < or = 0.06 microgram/ml) and rifampicin (MIC < or = 0.5 microgram/ml). All the strains belonged to serogroup C serotype 2b, serosubtype P1.2,5. During the study period 4 additional cases of meningococcal disease by serogroup B were observed. PMID- 9410051 TI - [Intensive care for patients with the human immunodeficiency virus]. AB - BACKGROUND: HIV infection is of special importance in Spain, occasionally requiring the use of intensive care units (ICU) in the management of these cases. METHODS: A retrospective review of the seropositive HIV patients was carried out of those requiring ICU admission in the authors' center over a three year period. Twenty-five patients were included, with the cause for admission into ICU, the clinical features, mortality and prognostic factors being reported. RESULTS: Most of the patients presented as a risk behavior by intravenous drug addiction and did present AIDS prior to hospitalization. The most frequent cause for admission in the ICU was respiratory failure produced by germs other than Pneumocytis carinii, placing this microorganism in second place. The mean time of admission in the ICU was 10.8 days, with 88% requiring mechanical ventilation and 56% the use of vasoactive drugs. The ICU mortality was 48%. No survivor died posteriorly on the ward. Factors of bad prognosis were: less than 200 CD4 lymphocytes/mm3, low albumin levels and high creatinine in addition to the need for vasoactive drugs with no evidence of other data presenting statistical significance. Fourty four percent of the survivors remain alive at 12 months of hospital discharge. CONCLUSIONS: Respiratory failure produced by Pneumocystis carinii or by other germs is the most frequent cause for ICU admission of patients with HIV infection. Renal failure, lymphopenia, and hypoalbuminemia are predictors of mortality in almost 50% of the cases. PMID- 9410052 TI - [Endophthalmitis in a patient from a rural environment]. PMID- 9410053 TI - [Female with erythematous papular lesions of the leg]. PMID- 9410054 TI - [Vibrios of marine origin in human pathology]. PMID- 9410055 TI - [Fluconazole treatment of hepatic abscesses caused by Candida albicans]. PMID- 9410056 TI - [Bacteremia caused by Shewanella putrefaciens]. PMID- 9410058 TI - [Disabling pelvic osteomyelitis caused by Echinococcus granulosus]. PMID- 9410057 TI - [Osteomyelitis caused by Eikenella corrodens]. PMID- 9410059 TI - [HIV infection and porphyria cutanea tarda]. PMID- 9410060 TI - [Isolation of Alternaria alternata in peritoneal fluid in patient in ambulatory chronic peritoneal dialysis]. PMID- 9410061 TI - [Aortic endocarditis after implantation of a provisional pacemaker]. PMID- 9410062 TI - [Acute pericarditis caused by human parvovirus B-19]. PMID- 9410063 TI - [Penetration of meropenem in gram-negative bacilli. Differences in activity with imipenem]. AB - The outer membrane of gramnegative bacteria cell-wall contain channels formed by proteins known as porins, which facilitate the penetration of molecules into the cell. Imipenem and meropenem possess an important intrinsic activity against most gramnegative bacteria due to their high affinity for the penicillin-binding proteins PBP-2 and/or PBP-3. Meropenem is slightly more active against certain species of Enterobacteriaceae, Pseudomonas aeruginosa and nonfermenters bacilli. In this sense the differences in the activity between the two carbapenems may be attributable to differences in their affinity for PBPs, differences in their resistance to beta-lactamases hydrolysis, or to the differences in the capacity to employ certain porin channels. OprF is the main porin channel involved in the beta-lactam penetration of bacteria, though OprC and OprD2 may also contribute to the penetration of carbapenems into P. aeruginosa. However, evidences suggest that although impenem requires the presence of OprD2, meropenem may use other pathways for penetration. In Escherichia coli both carbapenems use ompF and ompC, and no specific porin channels have been detected. Only in the case of Enterobacter cloacae may exceptions exists among Enterobacteriaceae. PMID- 9410064 TI - [Meropenem: microbiologic perspective]. AB - Meropenem is a beta-lactamic carbapenem derived from thienamycin and is structurally characterized by the presence of a beta-methyl group in position C1 which confers stability to the molecule versus renal dehydropeptidase 1 (DHP-1), thereby making the coadministration of an enzyme inhibitor unnecessary. Its esterochemical configuration of the lateral chain in C2 (dimethyl carbomoilpyrrolidenethium) increases the activity versus gram negative bacteria (enterobacteria and pseudomonas) and moreover, may explain the reduction in the proconvulsive effect observed in imipenem/cilastatin. Meropenem has great bactericide power and has a very wide spectrum of activity depending on it low molecular weight and zwiterionic structure, stability versus almost all the clinically important beta-lactamases and high affinity for the PBPs. It covers gram positive aerobes (Staphylococcus aureus, coagulase negative staphylococci, streptococci including Streptococcus pneumoniae resistant to penicillin, Enterococcus faecalis, Rhodococcus equi, Listeria monocytogenes) and gram negative bacteria (enterobacteria, P. aeruginosa, Acinetobacter, Aeromonas, Plesiomonas, Vibrio, Haemophilus influenzae, Neisseria, Moraxella) and anaerobes (Bacteroides, Prevotella, Porphyromonas, Fusobacterium, Clostridium, Peptostreptococcus, and Propionibacterium acnes), being more active than imipenem versus gram negatives: P. aeruginosa (2-4-fold), enterobacteria (2-32-fold) and H. influenzae (4-8-fold) and less active versus the gram positives (enterococci, streptococci and staphylococci). Meropenem has no activity on Enterococcus faecium, S. aureus resistant to methycillin, Stenotrophomonas maltophilia and other genera producers of chromosomic methalo-beta-lactamases (carbapenemases). Resistance may be due to impermeability given the loss of the OprD porin (OprD2 in enterobacteria and P. aeruginosa) loss of different membrane proteins (Proteus mirabilis, Proteus rettgeri, Enterobacter cloacae, Enterobacter aerogenes), modifications of the PBPs (gram positive) and the production of carbapenemases (chromosomic methalo-beta-lactamases). PMID- 9410066 TI - [Meropenem against bacteria carriers of wide spectrum TEM beta-lactamases: evolutive aspects]. AB - The so-called wide spectrum beta-lactamases (WSBLs) are able to hydrolyze wide spectrum cephalosporins or monobactamics such as cefotaxime, ceftriaxone, ceftazidime, cefepime, cefpiroma or aztreonam. The natural wide spectrum beta lactamases are mutational variants of TEM-1 consisting in the substitution of one of more amino acids within seven well defined positions in the molecule. Given the expected extremely low frequency for the simultaneous production of double or triple mutations, it is plausible that one of the mutational changes has been independently selected. A plurimutational remodelling of the TEM-beta-lactamase molecule is successively produced with the consequent appearance of highly effective ESBLs. Mutagenesis techniques allow clean molecular variants to be produced and allow the mutational effects under homogeneous conditions of bacterial strain, the plasmid implicated or the genic promotor to be studied. Meropenem remains active versus all the wide spectrum beta-lactamases referred in the 2be group of Bush, Jacoby and Medeiros as well as the new beta-lactamases produced in vitro by directed mutagenesis. PMID- 9410065 TI - [in vitro activity of carbapenems against Enterobacteriaceae and Pseudomonas aeruginosa hyperproducers of group 1 chromosomal beta-lactamases]. AB - Resistance to imipenem and meropenem, reported sporadically in Enterobacteriaceae and more frequently in Pseudomonas aeruginosa, can be caused, among other mechanisms, by the combination of changes in permeability and hyperproduction of inducible chromosomal beta-lactamases. In this study, the in vitro activity of imipenem and meropenem was analysed by the agar dilution method against cefotaxime, ceftazidime, and aztreonam resistant clinical strains of Enterobacteriaceae (n = 202) and P. aeruginosa (n = 90). This phenotype is consistent with the hyperproduction of group 1 chromosomal beta-lactamases and was previously determined in stably derepressed mutants in the same species, obtained from strains with inducible beta-lactamase expression by selection with cefotaxime and ceftazidime. Likewise, the activity of imipenem and meropenem against the same number of clinical isolates susceptible to cefotaxime, ceftazidime, and aztreonam was evaluated. In general, imipenem and meropenem showed an excellent activity, which was intrinsically greater for meropenem against Enterobacteriaceae and P. aeruginosa organisms. Nevertheless, imipenem and meropenem activity was slightly affected on cefotaxime, ceftazidime, and aztreonam resistant isolates of E. cloacae (MIC90, 1 and 0.2 microgram/ml, respectively), E. aerogenes (1 and 0.2 microgram/ml), C. freundii (1 and 0.1 microgram/ml), M. morganii (1 and 0.5 microgram/ml), and S. marcescens (4 and 0.5 micrograms/ml). On the other hand, the activity of imipenem and meropenem against ceftazidime and aztreonam resistant isolates of P. aeruginosa was more significantly affected, with MIC90 values of 64 and 16 micrograms/ml, respectively. PMID- 9410067 TI - [Meropenem: pharmacologic advantages of clinical interest]. AB - The pharmacokinetic profile of meropenem is similar to that of imipenem/cilastatin. It differs in resistance to hydrolysis by the methaloenzyme, dehydropeptidase I, and therefore does not require the combination of cilastatin, being administered alone. Similar to other beta-lactamic drugs, it is mainly distributed in the extravascular space (apparent Vd, 21 I) with an half life of elimination of approximately one hour. A low proportion binds to plasma proteins (< 20%). The tissue concentrations are maintained for prolonged periods at values greater than the MIC of most pathogens. In LCR and aqueous humor it has limited penetration. Nonetheless, the levels achieved in patients with meningitis (40 mg/kg/8 h) range between 0.9 and 6.5 micrograms/ml, greater than the MIC of most pathogens associated with this disease. The renal clearance of meropenem surpasses that of creatinine, thereby indicating excretion by glomerular filtration and tubular secretion. The doses in patients with renal insufficiency should be reduced according to creatinine clearance. Modification of the dosage is not necessary in patients with hepatic failure. Administered at a doses of 1 g/8 hours meropenem presented a value of (AUICo infinity) 60-90 micrograms/h/ml. The (AUIC)24 = 125 ensures that the serum concentrations are maintained above 3 micrograms/ml during a period greater than 80% of the dosage interval. PMID- 9410068 TI - [Clinical pharmacology and indications of meropenem in severe pediatric infection]. AB - The choice of an antibiotic for the treatment of a serious paediatric infection is generally a difficult problem. The arrival of Carbapenem resulted an important advance in the field of medicine due to its broad-spectrum, low incidence of resistances and good safety profile. Among Carbapenems, Meropenem introduction represents a progress because of its pharmacokinetics characteristics and blood brain barrier penetration. Meropenem dosage depends on the patient weight, and the way of administration is potentially easier. Meropenem has been compared with the most used paediatric antimicrobial in controlled and randomised clinical trials, showing a high efficacy in the treatment of several infections (respiratory, urinary, intraabdominal, dermatological, septicemia) and in neutropenic and cystic fibrosis patients aged between one month and twelve years old. Meropenem is specially useful in the bacterial meningitis treatment because it penetrates into the cerebrospinal fluid of patients with inflamed meninges and reaches therapeutic concentrations, and because appearance of seizures is low. Adverse reactions produced by Meropenem show a poor incidence and its severity is usually mild. With regard to this characteristics, it can be concluded that Meropenem is an antimicrobial which efficacy and safety profiles guarantee its use in the treatment of severe paediatric infections. PMID- 9410069 TI - [Pharmacoeconomics of meropenem versus imipenem/cilastatin]. AB - The Pharmacy and Therapeutics Committee of our hospital regulates the use of antimicrobial drugs every year. With the coming into the market of the new antibiotic carbapenem meropenem, comparative meropenem, it was assigned to the Drug Information Center the elaboration of comparative report on the two antibiotics, including as well cost analysis. We have analyzed the activities to be made since an antimicrobial drug is purchased until it is administered. We have only assessed on the direct costs that differ in the two antibiotics that are: the pre-use preparation in the pharmacy service and the preparation and administration in hospital floors. We have classified the direct cost in: goods and services, purchases and supplies costs costs and staff costs. We have calculated the daily cost of both antibiotics in the treatment of serious infections and even in more serious, life threatening infections. On top of that, we have a reviewed the use of imipenem/cilastatin and meropenem in our hospital. The information obtained shows that the saving in serious infections is of 34.6% and of 4.5% in more serious infections with vital compromise. The Pharmacy and Therapeutic Commission taking into account the report elaborated, has considered convenient the choice of meropenem as a representative of carbapenem drugs. PMID- 9410070 TI - [Meropenem in the treatment of surgical intra-abdominal infections]. AB - Intraabdominal infections are severe with a high morbidity and mortality which may be produced by multiple causes: perforation of the empty viscera, intestinal inflammatory processes, vascular pictures, abdominal traumatisms as a consequence of surgery. From a microbiologic point of view, different types of gram positive and gram negative aerobes such as anaerobe microorganisms or fungi may be isolated as causes. Polymicrobial infections are usually observed. The main treatment policy is to correctly eliminate the causing foci of the bacterial contamination whether surgically of by percutaneous drainage. Parallelly, it is essential for the surgeon to use appropriate antibiotic treatment. Meropenem, a carbapenem, has a wide spectrum antibacterial activity which cover gram positive and gram negative aerobes in addition to anaerobes leading to scarce adverse reactions. All the above leads meropenem to be a very effective alternative in both the treatment of these infections as monotherapy and for initiating empiric therapy. PMID- 9410071 TI - [The role of meropenem in bacterial meningitis]. AB - Meropenem is a new carbapenem antibiotic of with an antibacterian spectrum similar to that of imipenem, but from which it may mainly be differentiated by the possibility of its administration at high doses and it has no demonstrated proconvulsive effect, properties which make it applicable in the treatment of bacterial meningitis. The clinical and experimental experience in the treatment of bacterial meningitis with this antibiotic is herein reviewed. It has been observed that the efficacy and safety of meropenem in meningitis caused by N. meningitidis, H. influenzae and pneumococci sensitive to penicillin may be similar to that of cefotaxime or ceftriaxone in both the pediatric and adult population. There are very few reports on the treatment of meningitis caused by pneumococci resistant to penicillin. However, given that the activity of meropenem on these pneumococci is similar to that of cefotaxime and that the doses administered are much lower, it does not appear to be recommendable in the treatment of this indication, although it should be tested in all meningeal strain to these characteristics isolated. It may currently be considered that the main indication of meropenem in the infections of the central nervous system is in nosocomial meningitis by multiresistant gram negative bacilli such as those of the Klebsiella-Serratia-Enterobacter and Acinetobacter sp. group. Therefore a limited, albeit favorable, report on the clinical experience with meropenem is herein presented. Meropenem may also be useful in the treatment of meningitis by Pseudomonas aeruginosa in which other treatments have failed. PMID- 9410072 TI - [The role of meropenem in the treatment of pneumonia in the critical patient]. AB - Pneumonia in critical ill patients, most of them associated with insaturation of an artificial a way and the use of mechanical ventilation, involves important morbi/mortality in the Intensive Care Units. Knowledge of pathogenesis, risk factors, and implicated microorganisms in developing of this major infectious complication, in the context of infections which rise in the critically ill patients, allow us to apply prophylaxis measures which could decrease its incidence, and establish antimicrobial therapy, which permit us to cover all the etiologic possibilities. Availability in the arsenal of the powerful antimicrobial, of a new carbapenemic, meropenem, and based on the different studies and clinic assays, allow to recommend its use with warranties and efficacy, in the empirical or's in concretely those due to Pseudomonas aeruginosa, enterobacteriaceae, (in general or producers of ample spectrum beta lactamases), and Acinetobacter spp., in monotherapy or combined therapy, with aminoglycosides. PMID- 9410073 TI - [Monotherapy with meropenem in febrile granulocytopenic patients]. AB - Infection remains the major cause of morbidity and mortality for cancer patients who become granulocytopenic. Combinations of beta-lactams plus aminoglycosides have been the standard empiric therapy for febrile granulocytopenic patients, especially those with profound long-lasting granulocytopenia. The advent of new broad-spectrum cephalosporins and carbapenems has favoured the possibility of empiric monotherapy. Meropenem is a parenteral carbapenem antibiotic stable to renal dehydropeptidase-I which has excellent bactericidal activity against almost all clinically significant aerobic and anaerobic organisms. Meropenem hasta an antibacterial spectrum similar to that of imipenem but it is more active against Pseudomonas aeruginosa, all Enterobacteriaceae, Haemophilus influenzae, Proteus spp, Morganella spp and Providencia spp. Recently, the efficacy, safety, and tolerance of meropenem monotherapy for the empirical treatment of fever in granulocytopenic cancer patients have been compared in two large prospective randomized multicenter trials. The Meropenem Study Group compared monotherapy with meropenem versus ceftazidime and the EORTC conducted a comparative study of meropenem monotherapy versus the combination of ceftazidime plus amikacin. In both groups, success rates were similar by type of infection and infection related mortality was low. Related adverse events were also similar in both groups. These studies confirm that monotherapy with meropenem is as effective as ceftazidime-containing regimens for the empiric treatment of fever in granulocytopenic patients. PMID- 9410074 TI - [Tolerance and safety of carbapenems: the use of meropenem]. AB - The purpose of this article is to review the safety and tolerance of two carbapenems (imipenem/cilastatin and meropenem) in order to establish their possible use in different clinical settings. The tolerance and safety profile of both carbapemens in intravenous and intramuscular formulation is good. With imipenem/cilastatin, nausea and vomiting can constitute a practical problem requiring prolonged times of perfusion and high dilutions. The possibility of administering meropenem in intravenous infusion or bolus injection with lower volumes of fluid, without increasing the incidence of these adverse reactions, may have practical advantages in special situations. The possible neurotoxicity of the imipenem/cilastatin presents limitations of the use in high risk circumstances such as meningitis, previous alterations of CNS, renal insufficiency and concomitant administration of other drugs with neurotoxic profiles and when high doses of administration are needed. The meropenem, by the contrary, can be used in patients with infections of the CNS and other risk factors, at high doses, without increased risk of seizures. PMID- 9410075 TI - [The role of carbapenems in the treatment of nosocomial infection]. AB - Carbapenems are active beta-lactam antibiotics versus most of the gram positive and gram negative microorganisms and anaerobes although their activity is lacking in the case of Staphylococcus sp. resistant to methicillin, Enterococcus faecium and Streptococcus pneumoniae with high resistance to penicillin and some gram negative bacilli which naturally produce an methaloenzyme able to hydrolyze them such as Stenotrophomonas maltophilia. Imipenem, the first synthetized carbapenem requires administration with cilastatin to avoid inactivation by renal dehydropeptidase 1. Meropenem does not require being taken with the renal enzyme inhibitor, with its activity being similar to that of imipenem. In abdominal infection the carbapenems have shown to be the authentic monotherapy in this type of infections being as effective as the different schedules of antibiotic associations normally used. Treatment with carbapenems in bacterial meningitis should be currently limited to the cases produced by gram negative bacilli producers of wide spectrum beta-lactamases (WSBL), cases of meningitis by Pseudomonas aeruginosa or gram negative bacilli producers of inducible cephalosporinase. Meropenem is the carbapenem of choice probably in these cases because the carbapenems are often the only active antibiotics and meropenem, specifically, does not have the risk of convulsions observed with imipenem cilastatin. The carbapenems have shown to be useful in skin and soft tissue infections as well as in obstetric and gynecologic infections as monotherapy similar to the schedules of the currently used antibiotic associations. In the case of nosocomial pneumonias, all the studies have evaluated the carbapenems in monotherapy as useful and effective, specially in the case of pneumonia by gram negative bacilli. Finally, in non filiated nosocomial sepsis and specially in the case of neutropenic patients, the use of carbapenems is particularly attractive in gram negative sepsis in intensive care units. The appearance in the last few years of strains of gram negative bacilli, producers of wide spectrum beta lactamase or stable repressed hyperproducers of class I chromosomic cephalosporinase, as well as other multiresistant gram negative bacilli, such as Acinetobacter baumanii make the carbapenems, in many cases, the only effective antibiotic in this type of infections. PMID- 9410076 TI - [Anti-anaerobic activity of carbapenems]. AB - The critical concentrations of sensitivity and resistance for meropenem versus anaerobic bacterias were analyzed. It is demonstrated the meropenem is a powerful inhibitor of the microorganisms of the Bacteroides groups (MIC90 < 1 mg/l), Prevotella > Porphyromonas, Fusobacterium, Veillonella, Clostridium perfringens is inhibited at a MIC < 0.06 mg/l. The MIC of meropenem versus C. difficile is 2 mg/l. They are also highly susceptible to the Propionibacterium, Peptostreptococcus and Peptococcus type microorganisms. Meropenem is comparable to imipenem and is more active than piperacillin, metronidazole and clindamycin. The mechanism of action and acquisition of resistance versus meropenem is evaluated and discussed. The percentage of highly resistant strains (MIC > 256 mg/l) isolated in the Hospital Gregorio Maranon in Madrid (Spain) is low (1.2%). The influence of pH on the in vitro activity of the carbapenemicos is also analyzed providing experimental data suggesting that meropenem maintains its bactericide activity more effectively in low pH conditions (5.6). Finally, the authors analyze the literature and the evidence reported regarding the use of meropenem in clinical practice, as a treatment of intraabdominal infections with a clinical response of 91%-100% and bacteriologic efficacy of 84%-95%. PMID- 9410077 TI - [Septic shock and nitric oxide]. AB - Refractory hypotension is the main cause of death of patients with septic shock. It has been shown that an excessive release of NO is responsible for the sepsis induced hypotension and vascular hyporeactivity. Nitric oxide is produced under normal conditions by a constitutive enzyme present, among other cell types, in the endothelial cell, and is necessary for maintenance of normal organ perfusion. Under inflammatory or septic conditions, a new enzyme is expressed in phagocytic cells and vascular smooth muscle cells, giving rise to an uncontrolled NO production that is associated with cytotoxic effects and vasodilatation. Randomized clinical trials have shown that the administration of inhibitors of NO synthesis to patients with septic shock is associated with a greater incidence of shock resolution, without significant adverse effects. The recent discovery of the different biological functions of NO, both under normal and inflammatory conditions, has allowed the development of new concepts about the pathophysiology of septic shock, and has provided the bases to design novel therapeutic strategies for the treatment of septic shock, based on the inhibition of NO synthesis. PMID- 9410078 TI - [Diagnosis of pneumonia in intubated patients: a controversy without resolution?]. AB - There is yet no accurate, fast, innocuous and inexpensive method for the diagnosis of pneumonia associated to ventilation (NAV). Here we analyse three diagnostic lines of increasing level of certainty. CLINICA METHOD: Associates radiological image, pus in the trachea and temperature changes and or leukocytosis. Although it is quite sensitive in the absence of distress, is it not very specific. Nevertheless, it must be appreciated for its function as a guide, recalling that in its absence bacterial counts are of scarce value. QUANTITATIVE BACTERIOLOGICAL STUDIES OF DEEP SPECIMENS: Broncho-alveolar lavage (BAL) and protected brush (PB) serve a double objective: to discriminate colonisation from infection, and the identification of the etiological agent. The sensitivity of BAL/PB is approximately 70% while specificity is close to 80%. The method performs better in the absence of previous antibiotic therapy. HISTOLOGICAL DIAGNOSIS: Almost never obtained while the patient is alive, it is nevertheless the golden standard. The absolute diagnosis of NAV is: histological for pneumonia, with a positive tissue culture. USE OF DIAGNOSIS IN THERAPY: Empirical treatment results in a 40% failure rate. The specific treatment requires therapeutic changes in more than 1/3 of cases. Treatment when the micro organism is known is followed by longer survival. Perhaps the correct approach would be: early empirical treatment after taking deep samples and correction according to the results obtained. PMID- 9410079 TI - [Postoperative infections in critically ill patients]. AB - Patients subjected to surgery often develop nosocomial infections, among which the intra-abdominal ones stand out as being a common cause of septicemia, multi organ failure, and death of the critical patients. Advances have been made in the study of the physiopathology by studying the mediators which are responsible for the systemic inflammatory response, the microbiology (changes in the pathogen type and in the antimicrobial sensitivity), and for the clinical picture (cholecystitis, tertiary peritonitis). Abdominal ultra-sound and computerized axial tomography have contributed greatly to the diagnosis of these infections. The new treatment techniques are discussed, both of the drainage of the septic focus (percutaneous or surgical), as of the antimicrobial treatment and the supportive measures. The diagnostic and therapeutic advances have modified the prognosis of these patients, although this continues to be poor when there is development of the multi-organ failure syndrome. PMID- 9410081 TI - [Antibiotic policy in intensive care]. AB - The use of antibiotics takes places by following a series of norms, which are called the antibiotics policy. The importance of their correct use is based on the influence which the antimicrobial agents may have on the development of resistance by the endogenous agents in a specific hospital or department, and on the other hand, by their capacity to predispose patients for colonization with resistant flora. This is specially important in critical patients who are hospitalized in the Intensive Medicine Department, where the use of antibiotics affects more than 50% of those admitted, and where the appearance of multiresistant pathogens is frequent. The choice of the antibiotics which are used for the empirical and/or focused treatment of the majority of infections, is the basis of the antibiotics policy. It is based on the knowledge of the there essential pillars of the problem: the pathogenic agents which are predominant in each area, the patients' characteristics which favor the appearance of specific micro-organisms, and the antibiotics which can be used for their treatment. In the hospital, the establishment diffusion, and enforcement of these rules, is conducted by a multi-disciplinary committee, in which an intensive care physician specialized in infectious pathology participates, in close collaboration with other specialists (microbiology, pharmacy, infectious diseases), all of whom are committed to the control and correct use of antibiotics. PMID- 9410080 TI - [Catheter infections: before and after the Consensus Conference]. AB - The use of intravascular devices in the usual and daily clinical practice, used as diagnostic or as therapeutic means, is a reality nowadays, present in all hospitals of our medium, and it is increasing. In the area of intensive care, it constitutes a routine and prolonged procedure, and its contribution within the hospitality area, in the context of complications derived from its use, is important. The most frequent complication derived from the use of intravascular catheters or devices, is infection, which manifests itself either locally or systemically. At present it is considered that the bacteremia associated with a catheter, is by far the most common nosocomial cause of bacteremia in our environment, along with the associated morbido-mortality which this implies, the prolonged hospitalization, and the increased cost of health care. The connection of these catheters is an entry site which should be considered, and it is taking on greater significance in these past years. As always, the prophylaxis measures which are applied rigorously and on a daily manner, are the corner stone on which the possibility of minimizing these complications rests. PMID- 9410082 TI - [Severe community-acquired pneumonia]. AB - There is no precise definition for severe community-acquired pneumonia (SEHP), but there are a number of factors which are associated with a greater severity, and which therefore recommended the admission of these patients in an intensive care unit (ICU). In the present article, we mainly refer to SEHP in the immune competent population. SEHP makes up 8-10% of the total number of admissions of an ICU although this depends greatly on the type of unit concerned. The majority of patients admitted, do so because they need mechanical ventilation, because they present a shock situation, or because they develop a multi-organ failure in the course of the disease. The battery of usual tests recommended basically includes a chest x-ray, arterial gas, a count of red and white blood cells, biochemical profile, blood cultures, analysis and culture of the pleural liquid (if this is present), and respiratory samples. The therapeutic strategies tend to guarantee the simultaneous coverage of S. pneumoniae, H. influenzae, and the so-called atypical pathogens. Keeping in mind the considerable percentage of penicillin resistant pneumococcus existing in our country, in a general manner it is recommended to use a combination of a macrolide with a 3rd generation cephalosporin against this organism. They should be detected early, especially those situations in which there is respiratory failure and shock which shall require the use of mechanical ventilation and inotropics as well as an adequate monitoring. PMID- 9410083 TI - [Surveillance and control of infections in the intensive care unit: rates, resistance, and carrier state]. AB - The nosocomial infection detection systems in Intensive Care Units, present certain peculiarities. The traditional methods, the most important example of which is the National Nosocomial Infections System, have, among their most explicit main objectives, the comparison of the number of infections, and the infectious flora between Units, and within the same Unit, over a period of time. For this reason, they are experiencing a great development within the hospitality quality control programs. In the last decade, the control of the flora of the digestive apparatus, the oropharynx, and the rectum of ICU patients, has permitted the making of an etiopathogenic classification of the infections (primary endogenous, secondary endogenous, and exogenous), which has justified the use of preventive measures with topical and systemic antimicrobial agents, according to the cases, with a tremendous impact on the decrease in the number of pneumonia's associated with mechanical ventilation, and of early pneumonia's which traditionally are excluded from the traditional detectional systems, as well as the control of multiresistant gram-negative bacilli. For this reason, it is recommended that both methods be complementary to each other. Neither of the two detection systems has been evaluated in terms of efficiency. PMID- 9410084 TI - [Bacteremias in intensive care]. AB - Critical patients are exposed to monitorization and treatment systems which, in addition to the defense mechanism alterations which accompany their basic disease, favor their acquisition of nosocomial infections. Bacteremia's, along with respiratory infections, are the main nosocomial infections seen in Intensive Care Units (ICU's). Between 20 and 30% of the nosocomial bacteremia's occur in ICU's, which represents an incidence rate which varies from 2.5 to 6.7 episodes per 100 admissions, with these occasionally being epidemic type infections. The main origins are intravascular catheters, and respiratory infections, and in 50% of the cases, Gram positive organisms are responsible. The Gram negative bacteremia group is dominated by Pseudomonas spp., and by other non-fermenting Gram negative bacteria. Also, the incidence of candidemia is higher than in case acquired in conventional hospitalization units. The global mortality is set between 30 and 40%, although the mortality directly associated with infection is close to 70%, which confirms the importance of the underlying diseases in the final prognosis of nosocomial infections in critical patients. PMID- 9410085 TI - [Lipid peroxidation in critical patients]. AB - Lipid peroxidation is a part of normal metabolism. The degree of peroxidation of membrane depends on the quantity and the quality of his fatty acids, both as components of the membrane, as constituents of the cellular environment. The magnitude of the changes which result in response to an exogenous load of hydrogen peroxides, shall depend, in part, on the pre-existing tissue levels of endogenous hydrogen peroxide. The exposure of the membrane to oxygen free radicals produced in excess, stimulates the process of lipid peroxidation. An update is made on lipid peroxidation in different nosological entities, with special emphasis on the special context of critical patients, and within this group, on the concept of ischemia-reperfusion. On the other hand, a brief review is made of the possible antioxidant defenses which may be used in the clinical situation, and a more extensive review is made of the correct analytical methodology to be used for determining the peroxidation. PMID- 9410086 TI - [Acute respiratory distress syndrome. Nutritional and metabolic support]. AB - The application of artificial nutritional treatment in critical patients, nowadays is a completely accepted fact. To a large degree this is due to the advances made in the understanding of the metabolic response shown by patients against a severe and persistent aggression. One of the entities which presents the highest mortality in critical patients, is the Acute Respiratory Distress Syndrome (ARDS), which one understands as the pulmonary response to different types of aggression. To understand the metabolic implications in the face of the development of this syndrome, would permit a better understanding of the need to treat these patients and to establish the nutritional standards which are most adequate to each different metabolic alteration. It would not only be important to understand the most effective method for calculating the caloric needs of these patients, but we shall also have to deepen our understanding of possible harmful effects of the different substrates, which undoubtedly could condition morbidity. This is why in this review we focus on the intimate pulmonary mechanism, both in healthy conditions as in those of disease, in order to extrapolate conclusions which are potentially applicable to patients suffering from an acute pulmonary lesion, as those suffering from ARDS: A main role shall be played the assessment of the administration of macronutrients in the form carbohydrates, lipids, and amino acid particles, due to the implications which each of these may have on the lesioned lung and its ventilatory capacity. PMID- 9410087 TI - [Serum status of carotenoids in control subjects and its relation to the diet]. AB - Carotenoids are a group of fat soluble pigments which are present in the human being, both in blood, as in tissues, and which are obtained through the diet, mainly from fruits and vegetables. The interest of these compounds is due not only to the provitamin A activity of some of them, but also due to a whole series of biological activities such as: antioxidant or prooxidant, photo-protective, modulator of the immune response, anti-carcinogen, etc. The best analytical method available for the analysis of carotenoids is high performance liquid chromatography (HPLC), which is used in our study both for serum as for foods, and it is controlled throughout periodic quality controls. In this article we present the preliminary results of the levels of the major serum carotenoids (b carotene, a-carotene, b-cryptoxanthin, lutein, zeaxanthin, and lycopene) in control subjects from five European countries, as well as indicating the major dietary contributors to the carotenoids intake in the Spanish population. The percentage of each carotenoid to the total of the carotenoids analyzed, varies according to the origin of the studied population. Ireland and the UK show a very similar carotenoids profile. France presents the highest levels of lutein and b carotene, which are present simultaneously in green vegetables. Spain shows the lowest levels of b-carotene, along with the highest levels of b-cryptoxanthin, which in our country is supplied mainly by oranges and tangerines. The most abundant carotenoid in all countries was lycopene. The average daily intake of these carotenoids (from fresh fruits and vegetables) in our population, is 3.5 mg/day. Through the relationships between the dietary carotenoid contents and serum the identification of "biomarkers" have been proposed, which might be correlated with several pathological situations, and thus contribute to the prevention of certain diseases. PMID- 9410088 TI - [Oxygen metabolism in isolated rat hepatocytes in obesity. Influence of vitamin C]. AB - Nutritional obesity induced by the ingestion of hyperlipidic diet implies a high consume of lipids, which might be involved in oxygen metabolism. Double bound, in the fatty-acid molecules are a vulnerable point to undergo oxidation reactions generating lipid peroxidation, that are potentially toxic and can produce serious cell injury (alteration in cell permeability and prostaglandins...). To prevent this oxidative injury the aerobic organisms have intracellular mechanisms of defense, the antioxidant systems, that may be classified as enzymatic and nonenzymatic. Vitamin C belongs to the second group and acts as scavenger of free radicals and other species. The purpose of this study was to evaluate the oxygen metabolism in isolated hepatocytes of rats which obesity has been reached by the ingestion of an hyperenergetic olive-oil rich controlled liquid diet and evaluate the effect of ascorbic acid. Cellular oxidative injury in isolated hepatocytes was induced through a lipid peroxidative and cytotoxic molecule tert-butyl hydroperoxide (t-BOOH). Results show higher levels of lactate dehydrogenase (LDH) leakage and malondialdehyde (MDA) production in obese rats as compared with controls. Otherwise, when these groups are supplemented with ascorbic acid these changes decrease significantly. ATP levels decrease in hepatocytes of obese rats incubated in the presence of 1 mM tert-butylhydroperoxide. While it is maintained in ascorbic acid supplemented animals. GSH values were lower in hepatocytes from obese and control rats, incubated with tert-butyl-hydroperoxide. Supplementation with ascorbic acid also maintained GSH levels thus indicating that ascorbic acid is acting as an efficient antioxidant. PMID- 9410089 TI - [Nutritional parameters in long-stay critical patients]. AB - A study is made of the evolution of the nutritional biochemical parameters, albumin, prealbumin, cholesterol, creatinine index/height and transferrin, as well as the nutrition route, SAPS, APACHE II, chronic age score, and maximum degree of metabolic stress reached, involving all patients requiring artificial nutrition during at least 14 days, admitted to our intensive medicine unit during an 18 months period, with the aim of finding differences between survivors and those who died. The following conclusions were reached: 1) In patients with severe metabolic stress, like those of the present study, artificial nutrition manages to maintain the nutritional parameters within the limits of moderate malnutrition, improving the nitrogenation balance, without achieving its balance not reducing the consumption of lean body mass, represented by the progressive and significant reduction of the ICALT. 2) In our series, the nutritional parameters behave in a notably different manner with regard to the evolution. In survivors, improvements are seen in albumin, cholesterol, and prealbumin, without variations in transferrin, these changes not being seen in those who died, the latter also showing a significant drop in transferrin, and 3) The greater age and poorer prior health status, despite a lower APS, of those who died appears to be the determining factors for the mortality, and probably also for the different evolution of the nutritional parameters for the usual nutritional standards, maybe due to a lower response capacity to stress. PMID- 9410090 TI - [Nutritional support in bone marrow transplantation]. AB - Bone marrow transplant (BMT) implies the treatment with substances which may compromise the nutritional condition, thus increasing the morbido-mortality of these patients. The objective of this study is to evaluate the efficacy of the nutritional support (NS) protocol for patients subjected to a BMT in our center. PATIENTS AND METHODS: 55 patients were included (24 men and 31 women), who were subjected to BMT during 1994, with prior chemotherapy depending on the underlying disease. The nutritional condition (NC) was evaluated upon initiation and at the end of the NS, using anthropometric, biochemical, and immunological parameters. The NS was given by total parenteral nutrition (TPN), adapted to the needs, as of the second post-transplant day, until such time that oral nutrition was sufficient to supply the nutritional needs of the patients; oral ingestion was permitted at all times, according to the possibilities of the patient. For the statistical analysis, we used the Student's t test, Pearson's Chi squared test, and Spearman's test, with differences being considered significant for values < 0.05. RESULTS: The average duration of the TPN was 16 +/- 6 days, with a significantly longer time (p < 0.05) in patients with leukemia. The NC assessment was no different at the beginning and at the end of the NS, although all groups show a drop in the albumin levels at the end with respect to those at the beginning, with this being statistically significant in patients with leukemia (p < 0.05), and with solid tumors (p < 0.01), 14.5% of the patients maintained an acceptable oral ingestion (with 75% having lymphomas), and 34.5% did not show any associated oral ingestion. Te better albumin maintenance was correlated with acceptable oral ingestion (p < 0.05). CONCLUSIONS: Nutritional support of patients subjected to a BMT is effective for maintaining their NC levels. The longest duration of the TPN, the lowest frequency of associated oral ingestion, and the greatest decrease of the serum albumin, levels are seen in those cases which had the most aggressive chemotherapy prior to the BMT, which requires adaptation of the NS in function of the underlying disease. The association of oral ingestion may be beneficial due to its effect on the gastrointestinal tract. PMID- 9410092 TI - Evidence and experience: we still need opinions and hypotheses. PMID- 9410091 TI - [Physico-chemical characteristics of different types of vegetable fats and oils used in the manufacture of candies]. AB - The quality of three vegetable fats (cocoa butter and two commercial fats) and three roasted nut oils (almond, hazelnut and peanut) used as raw material in the chocolate products manufacturing was studied. The hydroperoxide content, oxidative stability and fatty acid composition were determined and its health repercussion by atherogenicity and thrombogenicity indexes. Two commercial fats and cocoa butter showed higher oxidative stability, atherogenic and thrombogenic properties than oils because of its different fatty acid profiles. Peroxide value was a low reliability parameter of raw material shelf live. Rancimat presented a good correlation with the unsaturation index of different fats and oils, it was a better index than peroxide value. In the chocolate products manufacturing it would be advisable a good raw material selection and formulation in order to get a balance between technological properties, organoleptic qualities and the influence on the health. Those raw material with less primary oxidation and higher oxidative stability were also those of higher atherogenicity and thrombogenicity indexes. PMID- 9410094 TI - Dresden Chemiluminescence Days. Dresden, 8-11 October 1997. Abstracts. PMID- 9410093 TI - Molecular Diagnostics Joint Congress of the German Society of Clinical Chemistry and German Society of Laboratory Medicine. Munster, 30 September-2 October 1997. Abstracts. PMID- 9410095 TI - Photo quiz. Tinea barbae caused by Trichophyton verrucosum. PMID- 9410096 TI - [Drug treatment of obesity: present and future]. PMID- 9410097 TI - [Weight-loss drugs: composition of diet pills prescribed in Navarra]. AB - BACKGROUND: The prescription, dispensation and sale of "Magisterial formulas" for weight loss is a very extended practice in Spain, in spite of being strongly unadvised by different Administration Department as well as by medical and sanitary groups. The composition of these preparations is not labelled if so only in a generic way. METHODS: The content of 54 capsules from 6 different origins (3 medical consultations, 1 pharmacist consultation, 1 free sale in pharmacies and 1 free sale in stores) was analysed in 2 laboratories. The presence and quantity of 29 active principles was searched by spectrophotometry, chromatography and radioimmunoassay. RESULTS: The presence of some of the 12 following substances was confirmed in 42 samples (77.8%): hormones (levothyroxine, cortisol and cortisone), appetite suppression phenetilamines (amphetamine, amphepranone, fenfluramine and fenproporex), benzodiazepines (diazepam, clorazepato and chlordiazepoxide) and diuretics (triamterene and ciclotiazide). The quantities of active principles was very variable in and among the six groups. In 13 of the capsules 1 active principle and in 29 combinations of two or more was found. None component of the labels, when available, corresponded with their analysed components. In 12 samples (22.2%) none of the which may correspond to some substances different from the 29 sought ones. CONCLUSIONS: The usage of formulas as the studied here on the overweight and obesity treatment should be obviously unadvised. PMID- 9410098 TI - [Frequency and characteristics of patients treated with zidovudine and absence of progression of HIV infection]. AB - BACKGROUND: the efficacy of zidovudine (ZDV) is lost in many treated individuals after a few weeks or months of monotherapy. The development of drug resistance seems to explain this adverse event. However, some individuals seem to persistently benefit clinically and immunologically from ongoing ZDV monotherapy. The degree and causes of this phenomenon remain unclear. PATIENTS AND METHODS: we studied 280 HIV-infected patients who have been receiving ZDV monotherapy for more than 18 months (mean 28 +/- 7 months), and whom has a CD4+ count between 200 and 500 x 10(6)/l at baseline. We classified them into two groups: Non progressors with ZDV (NP-ZDV), subjects with an increase or a reduction < 15% in the CD4+ count; and Progressors with ZDV (P-ZDV), subjects showing a decline in the CD4 count > 15%. Epidemiological, immunological and virological features of each group were compared. RESULTS: the prevalence of NP-ZDV in this population was 15.7% (44/280). Age, gender, and risk behaviour were not significantly different in NP-ZDV and P-ZDV. Although the CD4/CD8 ratio, as well as the CD45R0/CD45RA ratio into the CD4+ subpopulation, were higher in NP-ZDV than in P ZDV, the values did not achieve statistical significance. Virological studies were performed on 36 (81.8%) NP-ZDV and 55 (23.3%) P-ZDV. Mean HIV-RNA titer was higher in P-ZDV than in NP-ZDV (8.4 x 10(4) vs. 1.5 x 10(3) copies/ml; p < 0.01). Virus isolation from circulating mononuclear cells was made more frequently in P ZDV than in NP-ZDV (90.9% vs. 81.5%), although it did not achieve statistical significance. The syncitium-inducing (SI) phenotype was detected in more than a quarter (27.3%) of P-ZDV but was absent in NP-ZDV (p < 0.01). The prevalence of RT mutations at codon 215 was much lower in NP-ZDV than in P-ZDV, and it showed a strong statistical significance (13.9% vs. 74.5%; p < 0.01). CONCLUSIONS: prolonged (> 2 years) lack of immunological and clinical progression can be observed in 15% of HIV-infected persons with mild immunosuppression, undertaking ZDV monotherapy. This effect seems to be associated with a characteristic virological profile, in which a low viral load, the absence of SI phenotype, and a lack of development of ZDV-resistance are the most relevant features. PMID- 9410099 TI - [Contribution of hepatic scintigraphy with Tc99m blood cells to the imaging diagnosis of cavernous hemangioma. Results in 48 patients]. AB - Cavernous hemangiomas are the most common benign tumours of the liver, being the liver the most frequent organ affected by this lesions. We report 48 patients who underwent 99mTc-red blood cell (RBC) scintigraphy, after ultrasound or tomographic appearance of cavernous hemangioma. Scintigraphy findings in 29 of 48 patients (60.4%) were typical of cavernous hemangioma, hemangioma-positives, after scintigraphy (all true-positives). On clinical, ultrasound and in one case after surgery (exploratory laparotomy) the follow up was positive for hemangioma. Specificity of 100%. In the remaining 19 patients (39.6%) where hemangioma negative lesions, 17 of 19 where true negatives and the remaining 2 where angiomas false-negatives, both of them were small lesions. The sensitivity for hemangiomas was 89.4%. To conclude with we would like to enhance the high specificity and sensitivity of 99mTc-red blood cell scintigraphy on the diagnosis of cavernous hepatic hemangioma (CHH). We highlight the utility of this specific imaging technique in the final diagnosis of CHH not requiring other invasive diagnosis tests, such as laparoscopy that in our study proved to be unnecessary. PMID- 9410100 TI - [Analysis of cost minimization of epidural anesthesia compared with general anesthesia in oncologic coloproctologic surgery]. AB - BACKGROUND: Combined general and epidural anaesthesia in abdominal surgery has shown, both, protective and no effect on final outcome. The aim of this study was to evaluate combined epidural and general anesthesia. METHODS: One hundred and eighty four patients, diagnosed of neoplastic process, in whom an elective procedure of coloproctologic resection and reconstruction was scheduled during the period between January-1993 and December 1994, were studied. In thirty consecutive patients a combined general-epidural anaesthesia (EA) was performed. These patients were compared to thirty general anaesthesia patients (GA), selected randomly from the same period. RESULTS: Both groups were comparable for demographic characteristics and for the type and duration of the surgical procedure. Red Blood Cells units transfused were 1.7 +/- 3 in the EA group and 1.4 +/- 1.9 in the GA group. After the operation, most of patients went to SICU. The length of the hospital stay was 13 +/- 6 days for GA group, while for EA group was .13 +/- 5. The hospital mortality for all operated patients (N = 184) was 1.1%, which were directly related to failure of surgical anastomosis. The need for mechanical ventilation and pulmonary complications were similar in both groups. When analyzing costs, EA group represented a value (pesetas) of 433,501 +/- 183,337 for GA group and 437,735 +/- 149,572 for EA group. CONCLUSIONS: As shown, in the actual context, we conclude that the anaesthetic technique did not have any influence on outcome or on cost. PMID- 9410101 TI - [Treatment with interferon of chronic active hepatitis in patients with HIV infection]. AB - Clinical records of 14, CD4 cell counts > 400/mm3, mild symptoms or asymptomatic, HIV infected patients and with chronic active hepatitis (identified by hepatic biopsy) were under review. Four of them were infected with HBV, 8 with HCV, 1 with HDV and other one with HBV + HCV + HDV. They were treated with alpha interferon for 6 months. Effectiveness was evaluated. It was found that in 4 (50%) of HCV infected patients transaminases raised normal value two of them remained with normal values at the end of review (22 and 48 months of follow-up). All HBV infected patients (4) normalized transaminases. Three of them lost HBeAg, that persisted through 38 months of follow-up. It was found too, whose did not improved with 6 months treatment did not benefit with a longer treatment. Therefore, HIV infected patients uncompromised (CD4+ > 400/mm3) and with chronic active hepatitis were benefited by interferon treatment (57%). Reversal of HBeAg was remarkable. PMID- 9410102 TI - [Pleural effusion with diffuse lung involvement and single subcutaneous nodule as first manifestation of atypical bronchial carcinoid tumor]. AB - Patient with atypical carcinoid with pleural effusion, diffuse pulmonary involvement and subcutaneous nodule, bone metastases and mediastinal metastases as the first presentation of the carcinoid. Atypical carcinoid is defined as a higher incidence of metastases, and a worse prognosis. PMID- 9410103 TI - [Mesenteric panniculitis in an 87-year-old female patient]. AB - We report a case of atypical presentation of mesenteric panniculitis in a 87 year old woman. The mesenteric panniculitis disease and the cases published are revised. Delirium as atypical disease presentations and especially dementia, are briefly analyzed. PMID- 9410104 TI - [Bronchial atresia and bronchogenic cyst in an adult]. AB - We present the case of an asymptomatic patient, with a right paracardiac mass discovered in a preoperative radiologic study. The bronchoscopy showed the intermedius bronchus atresia, finished in bottom of sack, with absence of middle and lower right lobes. With the chest computed tomographic scan, two lobulated contour mediastinal masses were seen. The patient was submitted to surgery, being the pathological findings consistent with bronchogenic cysts with atresia of the intermediarius bronchus. The patient evolved favourably after surgery. PMID- 9410105 TI - [Osteonecrosis in systemic lupus erythematosus. Report of 3 cases]. AB - We present three cases of patients with systemic lupus erythematosus (SLE) and osteonecrosis or avascular necrosis (AV). Although, the pathogenesis of osteonecrosis is controversial and multifactorial, the glucocorticoids therapy is the most important factor contributing to the lesion. We report the clinical presentation of the three patients. We comment the characteristics of AV, the diagnosis and the treatment of this uncommon complication in SLE patients. PMID- 9410106 TI - [Tuberculosis and HIV infection: pathogenesis (2nd of 3 parts)]. PMID- 9410107 TI - [Polyarthritis, meningitis, and bilateral psoas abscess caused by Staphylococcus aureus]. PMID- 9410108 TI - [Massive hemoptysis with fatal outcome. Infrequent course of pulmonary hydatidosis]. PMID- 9410109 TI - [Adrenal calcifications and subclinical adrenal hypofunction]. PMID- 9410110 TI - [Malignant peritoneal mesothelioma as late complication of radiotherapy for Hodgkin's disease]. PMID- 9410111 TI - [Pulmonary tuberculosis and lung cancer]. PMID- 9410112 TI - [Eosinophilia: initial presentation form of hypernephroma]. PMID- 9410113 TI - [Cecum perforation with fatal outcome in the course of Ogilvie's syndrome]. PMID- 9410114 TI - [Mildly dilated myocardiopathy]. PMID- 9410115 TI - [Mediterranean boutonneuse fever]. PMID- 9410116 TI - [Study of 86 cases of Mediterranean boutonneuse fever hospitalized at a university hospital]. AB - OBJECTIVES: To study the clinical manifestations of 86 patients with Mediterranean Boutonneuse Fever who were admitted in a University General Hospital. PATIENTS AND METHODS: Between 1986 and 1994 we studied retrospectively the clinical manifestations, evolution and complications of 86 patients with Mediterranean Boutonneuse Fever. Diagnosis was based on clinical and serological findings. RESULTS: We studied a total of 86 patients (64 males; 22 females) with a mean age of 55 years. 88% of cases were diagnosed between June and September, and 89% of them had contact with dogs. 53% of patients had an underlying disease. All patients presented with fever and a generalized erythematous rash. 60% of patients had a initial lesion (tache noire) especially in legs. All patients were treated with doxicycline during one week. 22% of patients had complications such as renal failure, respiratory failure, gastrointestinal bleeding and stroke. Old patients and those with underlying disease had severe complications with a higher significant frequency. No patients died. CONCLUSIONS: 22% of patients with Mediterranean Boutonneuse Fever, especially those with advanced age or underlying disease, who were admitted in the Hospital had severe complications. PMID- 9410117 TI - [Estrogen receptors and breast fibroadenoma with suspicious traits of malignancy]. AB - Fibroadenomas (FAD) are the most common breast tumors. Their clinical and morphological characteristics often vary, and endocrine factors has been implicated in their etiology. The development of a breast carcinoma overlying a FAD is very rare. An immunohistochemical study is made of the presence of estrogen receptor (ER) in a series of 21 FAD with clinical, mammographic or cytological suspect of malignancy. An intraoperative biopsy evaluation was made in all cases. In three cases FAD was associated to breast cancer (three lobular carcinomas-two infiltrating and one in situ; one case was also adjacent to an infiltrating ductal carcinoma). Two of these three fibroadenomas were ER positive. Globally, only 35% of the FAD evaluated proved ER-positive. Of all the parameters evaluated (age, tumor size, single or multiple presentation, history of breast carcinoma or oral contraceptives, and histological characteristics) no association was observed with respect to ER positivity, with the exception of patient age. Thus, ER-positive FAD were found in patients 5.4 years younger than the ER-negative women (ages 28.4 and 33.8 respectively). On the other hand, the mean age (46.6 years) of the patients with FAD and cancer was 10.2 years above the mean age (34.4 years) of the patients with FAD but no cancer; however, and despite this greater age, two of the three patients with both FAD and cancer exhibited ER positivity at FAD level. PMID- 9410118 TI - [Assessment of prognosis factors in the cure of Cushing's disease surgically treated via a +septotransphenoidal approach]. AB - INTRODUCTION: In Cushing's disease (CD) pituitary surgery or radiotherapy has been proposed by some authors, when plasmatic cortisol after surgery is not clearly low. AIM: To assess if the different prognostic factors, specially plasmatic cortisol seven days after surgery and/or hypocortisolism phase are predictive of the CD outcome. METHODS: From 1988, 11 women with CD underwent 13 transsphenoidal microsurgery, because two patients relapsed. The mean age of patients was 27 years (11-52). Plasmatic cortisol was measured seven days after pituitary surgery, and since 45 days, every three-six months, basal plasmatic cortisol and after ACTH and urinary free cortisol were determined. RESULTS: Follow-up evaluations ranged from 18-84 months (median, 38 months). After pituitary surgery in 13 cases the cumulative remission was 100%, two cases relapsed. In 10 cases plasmatic cortisol seven days after surgery was less than 137 nmol/l and in three cases higher than 137 nmol/l. Three cases did not presented hypocortisolism phase. The two patients who relapsed, one was after eight months of pituitary surgery an previously showed low plasmatic cortisol and the other relapse 25 months after pituitary surgery without low cortisol plasmatic levels. CONCLUSION: Remission in CD can happen either low or normal plasmatic cortisol levels seven days posttreatment or without hypocortisolism phase. Ours findings ascribe new importance to the different presentations after treatment of CD, and patients with these findings are not a risk for relapse and pituitary surgery or irradiation would not be early indicated. PMID- 9410119 TI - [Heat syndrome: the first 7 cases in Granada]. AB - We present the seven first cases with heatstroke in Granada provinces. Six patients were elderly man with passive heatstroke, the other was a young adult with active heatstroke. The clinic picture was evident: fever, hiperpnea, anhidrosis and mental impairment; there was not peculiar biochemic sign though the prognosis depended of the development of tubular kidney failure. We expose the main phisiopathologic therapeutic measure. PMID- 9410120 TI - [Course and prognosis of levels of CD4 molecules in HIV-infected patients]. AB - OBJECTIVE: To measure CD4 molecule concentration on lymphocyte surface. And observe its evolution during one year, in HIV infected patients. To assess the importance of this measure as a prognostic marker for disease progression, compared with the total number of CD4 lymphocytes. METHODS: Clinical and analytical data from 107 patients were collected; 64 fulfilled a one-year follow up. 45 were initially asymptomatic (ASN), 13 had AIDS-related complex (CRS) and 6 had AIDS. CD4 cell count was higher than 500 in 16, between 500 and 200 in 20, and lower than 200 in 28. CD4 molecule concentration in lymphocytes was measured with immunoassay capcellia CD4/CD8 (Pasteur Institute, Paris). During one year 12 patients progressed cunically. RESULTS: There was significant decrease of T4 lymphocytes in the following groups whole population, CRS, more than 500 (initially). There were significant decreases of CD4 molecules in the following groups: whole population, ASN, CRS, more than 500 (initially), between 500 and 200 (initially), the mean number of T4 lymphocytes was 140/mm3 in progressors versus 358 in non-progressors (p < 0.05). Initial CD4 molecule concentration was 3.87 pmol/l in progressors versus 11.50 pmol/l in non-progressors (p < 0.05). CONCLUSIONS: Both total number of T4 lymphocytes and CD4 molecule concentration decrease with time. However, in asymptomatic patients only CD4 molecule decrease reaches statistical signification. Both T4 lymphocyte number and CD4 molecule concentration were lower in progressors. Measurement of CD4 molecule concentration is a parameter of similar value with respect to T4 lymphocyte count, and with similar value as progression marker. PMID- 9410121 TI - [Spinal cord sarcoidosis: diagnostic and therapeutic problems]. AB - Three cases of sarcoidosis with spinal cord involvement are reported. The diagnosis in each case was very difficult, being mistaken with tuberculosis in two of the patients and with paraneoplasic myelopathy in the other one. Initial treatment was incorrect for this reason. MRI without contrast was normal, but showed the existence of lesions in two cases after gadolinium administration. Symptoms improved in two patients with prednisone, but they got worse after tapering dose. Spinal sarcoidosis is very rare, but its diagnosis most be considered and MRI with gadolinium should be performed in subacute or chronic myelopathies. Treatment with prednisone should be prolonged. PMID- 9410122 TI - [Tuberous sclerosis. Description of a family study]. AB - We present tuberous sclerosis affecting all the members of a family, now and for three consecutive generations (probably four). A fortuitous study of cutaneous disease in one member led to diagnosis in all of them. A systematic study detected important visceral implication in all cases, mainly neurological, despite the fact only one of then showed related symptoms. We discuss the epidemiological and diagnostic aspects of an illness, which is widely underdiagnosed even today, despite the fact that detection is by sight. PMID- 9410123 TI - [Cardiac tamponade as first clinical manifestation of small cell carcinoma]. AB - A 69 year old man was diagnosed of cardiac tamponade. At surgery a large hemopericardias was evacuated, cardiac and pericardial metastasis of an oat cell carcinoma was demonstrated. The patient died 4 weeks later. A review of the literature is done about patients with cardiac tamponade as the first manifestation of a neoplastic process. PMID- 9410124 TI - [Cardiopathy and human immunodeficiency virus infection: report of 2 cases]. AB - Two patients with immune deficiency virus infection and cardiac manifestation are presented. The first was admitted by encephalopathy. On the first days of hospital stay were observed episodes of supraventricular tachycardia with echocardiography normal. An initial improvement made us suppose the clinical context in relationship to toxoplasmosis. Subsequently the patient experienced neurological deterioration and pericardial effusion was evidenced. Therapeutic trials with ceftriaxone and tuberculostatics were useless. The postmortem pericardial aspiration showed signs of acute inflammation, but germs were not identified. The second case was admitted for longstanding fever. Because of epigastric pain an electrocardiogram was done, which revealed suggestive alterations of pericarditis. The echocardiography demonstrated the presence of pericardial effusion. The positive serology for toxoplasmosis, the bone marrow biopsy which showed reactivity with hemophagocytosis and the presence of myeloid metaplasia in the liver biopsy were the most relevant data. On the fifth day of hospital stay and second of the tuberculostatic treatment the patient expired unexpectedly. PMID- 9410125 TI - [Usefulness of preoperative location tests in primary hyperparathyroidism]. AB - The utility of preoperative parathyroid location in primary hyperparathyroidism (pHPT) remains controversial. In this retrospective study are analysed noninvasive techniques prior to surgical neck exploration of 11 patients with pHPT, ten adenomas and one hyperplastic parathyroid glands. Localization test were performed 24 times. Ultrasonography allowed correct localization of 5 of 7 cases, computer tomography 2 of 6, scintigraphy 2 of 4 and nuclear magnetic resonance 1 of 7. This exploration was doubtful in a patient, and two false positive nuclear magnetic resonance were found. The low yield of these tests makes them unnecessary in the evaluation of patients undergoing surgery for pHPT, because the cost is not justified. PMID- 9410126 TI - [Tuberculosis and HIV infection: from clinical care to prevention (third of three parts)]. PMID- 9410127 TI - [Adverse reactions to insulin]. AB - The prevalence of allergic reactions to insuline has decreased during the last few years. Probably this is due to the use of the newly-developed recombinant human insuline. At present, adverse reactions to insuline occur in 5-10% of patients on therapy with insuline. Adverse reactions may be local (more frequent) or systemic (rare). Insuline resistance consists in a different type of immunological reaction. Diagnosis of allergy to insuline is based on clinical history and cutaneous and serological tests. Treatment depends upon the severity of the reaction. When insuline is indispensable despite a previous allergic reaction, a desensitization protocol may be implemented. PMID- 9410128 TI - [Endarteritis obliterans as a form of vasculitis in advanced rheumatoid arthritis]. PMID- 9410129 TI - [Dysphagia as presentation form of tracheal adenocarcinoma]. PMID- 9410131 TI - [Cancer. Terminal phase]. PMID- 9410130 TI - [Hypersensitivity vasculitis as first manifestation of multiple myeloma]. PMID- 9410132 TI - [Quality in medication prescription and evidence-based medicine]. PMID- 9410133 TI - [Pyomyositis at the root of the right thigh as presentation form of cecum adenocarcinoma]. PMID- 9410134 TI - [Pyogenic hepatic abscess; treatment with antibiotics and percutaneous drainage. Report of a case]. PMID- 9410135 TI - [Herpetic meningitis and cutaneous herpes zoster]. PMID- 9410136 TI - [Treatment of drug dependence at the health center: 10 years' experience]. PMID- 9410137 TI - [Puerperal depression. Related factors]. AB - OBJECTIVE: To find the prevalence of postnatal depression in puerperal women in the city of Albacete and to analyse its association with social and demographic factors, morbidity and social and affective support. DESIGN: An observational crossover study with a home interview. SETTING: Community. PATIENTS: 304 women who gave birth at Albacete General Hospital between May and November 1995. MEASUREMENTS AND MAIN RESULTS: The presence of possible depression between the sixth and eighth weeks after giving birth was measured with a self-administered test, the Edinburgh Postnatal Depression Scale (EPDS), on which scores of 13 or more are considered to signal depression. 15.8% (C.I. 11.8%-19.8%) of the women surveyed suffered a probable depressive disorder according to the EPDS. A significant association was found between puerperal depression and history of depression before pregnancy (p = 0.00001), during pregnancy (p = 0.00005) and immediately after delivery (p = 0.00001). Puerperal depression was also associated with stated chronic illness (p = 0.0003), scant help with domestic tasks (p = 0.003) and low family affective support (p = 0.0003). CONCLUSIONS: Between 11.8% and 19.8% of women between the sixth and eighth week of the puerperal period display a probable depression, which is linked to depression immediately after delivery. This disorder needs to be detected during the puerperal visit. PMID- 9410138 TI - [Chronic metabolic syndrome: a predictive model of risk of macrovascular events?]. AB - OBJECTIVES: To establish a predictive model of the risk of macrovascular complications in patients with Chronic Metabolic Syndrome by means of multiple logistic regression analysis. To identify Chronic Metabolic Syndrome as an independent health problem, given its frequency and importance in the genesis of macrovascular complications. DESIGN: A descriptive observational study. SETTING: An urban Health District in Malaga. PATIENTS: 47 patients with Chronic Metabolic Syndrome were chosen by systematic randomised sampling. MEASUREMENTS AND MAIN RESULTS: The best predictive model of macrovascular events was established as the one which included high values of the Waist-Hip index, blood pressure, Fibrinogenaemia and basal Glucaemia, and low values of HDL cholesterol. CONCLUSIONS: 1. A model to predict macrovascular events in patients with Chronic Metabolic Syndrome included high values of the Waist-Hip index, blood pressure, Fibrinogenaemia and basal Glucaemia, and low values of HDL cholesterol. 2. We believe that this association should be considered an independent health problem on the list of problems, with the name Syndrome X or Chronic Metabolic Syndrome. PMID- 9410139 TI - [Estimation of the prevalence of cognitive impairment according to the test used]. AB - OBJECTIVES: The aim of the study was to compare the findings of Folstein's Mini Mental Test (MMT), Lobo's Cognitive Mini Exam (CME) and the Isaacs Set test, in their detection of cognitive deficit in over-64 year olds; and to analyse the factors associated with variability in the scores obtained with these tests. DESIGN: An observational crossover study. SETTING: An urban Primary Care center. PATIENTS: Among the total of 1096 patients aged over 64 registered on 5 lists and with medical records, a random sample of 329 was chosen. INTERVENTIONS: The MMT, CME and Set test were administered to each patient in the same interview. Information on associated morbidity and social and demographic variables was gathered. MAIN RESULTS: 211 women (64%) and 118 men (36%) completed the tests. Women were older (73.0 +/- 6 against 71.2 +/- 6; p = 0.02) and had had less schooling (3.2 +/- 3.6 years against 4.2 +/- 3.7; p = 0.019). A higher percentage of women were illiterate (47.1% against 24.3%; p = 0.008). The MMT showed CD at 60.8%, the CME at 28% and the Set test at 11.3%. CONCLUSIONS: Estimated prevalence of CD varies depending on the test used. Variability in the MMT and CME is basically due to years of schooling, whereas in the Set test it is because of age. PMID- 9410140 TI - [Factors involved in noncompliance with pharmacological treatment in arterial hypertension]. AB - OBJECTIVES: To find the amount of non-compliance with medical treatment for Hypertension and its causes, and to describe the profile of non-compliant patients. DESIGN: A crossover study performed on two home visits. SETTING: A rural Health Centre at Calpe, Alicante. PATIENTS: The sample was obtained from the census of medically treated hypertense patients. 174 of the 200 patients chosen completed the study. MEASUREMENTS AND MAIN RESULTS: Compliance was evaluated by a surprise count of pills in the patient's home. Patients complying between 80 and 110% were considered compliant. There was 47.7% non-compliance (C.I. 95%: 40.3-55.1), with 31% under-compliers and 16.7% over-compliers. Lack of information (39.8%) and forgetfulness (28.9%) were the most common causes of non compliance. CONCLUSIONS: A high amount of non-compliance was shown, including an important number of over-compliers. Its causes were defined along with other reasons predicting non-compliance. PMID- 9410141 TI - [Self care and risk factors of diabetic foot in patients with type II diabetes mellitus]. AB - OBJECTIVES: To find the amount of self-care (SC), health education (HE) received and the prevalence of risk factors for diabetic foot (RFDF) in patients with type II Diabetes Mellitus (DM) attended in Primary Care. DESIGN: A descriptive crossover study. SETTING: Primary Care Centre. PATIENTS: 100 DM patients attending over 2 months (May and June 1995) to see the doctor or collect prescriptions. MEASUREMENTS AND MAIN RESULTS: Questionnaire on HE, SC habits and social and demographic data, inspection of the feet and physical investigation of lower extremities. 36% had deficient or very deficient hygiene; 73% did not go regularly to the chiropodist, 76% used scissors, 75% did not check the inside of the shoe. 38% had signs of neuropathy and 17%, of peripheric vasculopathy. 25% were at high risk of diabetic foot. Women had more RFDF. CONCLUSIONS: The amount of self-care is very low, especially in hygiene, which did not improve over time. HE on foot care is extremely poor despite its being a priority. Educational interventions are required to motivate healthworkers and patients, especially those with most RFDF, in the area of SC. PMID- 9410142 TI - [Are the indicators used to measure efficiency in primary care adequate?]. AB - OBJECTIVE: Getting to know the opinion of area managers and Primary Health Care Facilities coordinators about the indicators used to measure the PHC output. DESIGN: Questionnaire via mail. Descriptive statistics. SETTING: Areas 2-5, and 3, Zaragoza, Spain. PATIENTS: Managing teams of these three areas and Primary Health Care facilities coordinators. MEASUREMENTS AND MAIN RESULTS: The questionnaire was sent to 57 Primary Health Care Facilities coordinators and 16 area managers which was answered 53% of them. In the range from 1 to 10 the so called product achieved in the evaluation the highest marks have been: the Services offered by the PHCF (7.8), Coverage (7.8), Achievement of the Minimal Technical Rules (7.76) and Patient Satisfaction (7.51). The indicator that obtained the highest level of agreement was also the product achieved in the evaluation of the Services offered by PHCF. CONCLUSIONS: According to these results indicators normally used, related with number of visits are questioned by the professionals. Synthetic indexes recently introduced obtained better values. PMID- 9410143 TI - [Evaluation of the experience in domiciliary hospitalization of trauma patients]. AB - OBJECTIVE: To evaluate the use of "Hospital Care at home for trauma patients" from the perspective of decreasing the average hospital stay and termining the home care load engendered. DESIGN: Longitudinal descriptive study. SETTING: The Hospital Care at home unit of the Juan Canalejo hospital in La Coruna. PATIENTS: Patients belonging to the city of La Coruna or adjacent towns, who were operated on during the first six months of the programme for a trauma pathology (hip fracture, coxarthrosis, gonarthrosis). MEASUREMENTS AND MAIN RESULTS: The clinical records were reviewed. The following data were considered: number of patients, age, length of hospital stay, length of home follow-up, number of home medical and nursing visits required. CONCLUSIONS: This type of patient spends less time on average in hospital. PMID- 9410144 TI - [Stroke mortality in Andalusia: where are we going?]. AB - OBJECTIVE: The aim of the study was to explore temporal changes in stroke mortality in Andalusia over the period 1975-1994 and estimate the rates until the year 2000. DESIGN: Descriptive study. SETTING: Comunidad Andaluza. PATIENTS AND METHODS: The number of death from stroke was obtained for the period 1975-1994. Mortality raes were standardized directly using those of the "European population" as the standard, and regression analysis was undertaken. RESULTS: Age standardized overall mortality rates for stroke decreased considerably among men (45.6%). A similar pattern has been also observed among women (43.2%) during the period 1975-1992. If the trends continue during the period 1992-2000 we will observe an important reduction in both men (23.4%) and women (24.9%). CONCLUSIONS: In conclusion, this analysis shows a marked an steady fall in stroke mortality over the last two decades in Andalusia. Little is known about the factors that have led to the decline and further studies are necessary. A better understanding of these factors is important for planning the most effective intervention strategies to reduce, stroke mortality in our community. PMID- 9410145 TI - [Antibiotic treatment of otitis media in children]. PMID- 9410146 TI - [Social work students' knowledge about HIV]. PMID- 9410148 TI - [Knowledge of a particular group of diabetics about aspects of their diet]. PMID- 9410147 TI - [Acute post-influenza myositis in children]. PMID- 9410149 TI - [Whose are the true objectives?]. PMID- 9410150 TI - [Ethics and family and community medicine]. PMID- 9410151 TI - [Satisfaction of the "Mister Employee": a goal to be reached]. PMID- 9410152 TI - [Urinary difficulty caused by verapamil]. PMID- 9410153 TI - [Development of a computer-assisted thermoelectric Peltier cold test procedure with integrated photoplethysmography unit for noninvasive evaluation of acral skin circulation]. AB - Local cold provocation tests are an important, non-invasive diagnostic tool for collecting information about skin perfusion during exposure to cold. In patients suffering from vasospastic circulatory disorders such as Raynaud's phenomenon, it is of particular importance to be able to collect data about acral circulation during the cooling test in the asymptomatic intervals between naturally occurring attacks. By carrying out a series of cold provocation tests, for example, patient response to a newly initiated therapy can be assessed. Here we present a recently developed, computer-aided thermoelectric Peltier device with an integrated finger holder for carrying out local cold provocation tests. The electronic control unit of the Peltier element make it possible to cool or heat to predefined temperatures. At the same time, the temperature of both the finger holder and the skin can be measured. A photoplethysmographic sensor is also integrated within the device, enabling the response of the pulse waves to the controlled temperature changes to be monitored accurately. It is also possible to measure simultaneously laser Doppler flux and capillary pressure in the nailfold and to perform nailfold capillaroscopy to determine red blood cell velocity. The new device provides us with the technical means to study the interrelationship between acral skin perfusion and the thermal regulation of the skin. PMID- 9410154 TI - [Effect of boundary element discretization on forward calculation and the inverse problem in electroencephalography and magnetoencephalography]. AB - Modelling in magnetoencephalography (MEG) and electroencephalography (EEG) is increasingly based on the boundary element method (BEM). We quantify the influence of boundary element discretization on the neuromagnetic and neuroelectric forward and inverse problem for different dipole depths, brain regions and the quasispherical correction. In particular we derive standards for the general use of BEM models in MEG/EEG source localization. For this purpose simulation with single current dipoles, and source reconstructions from somatosensory evoked potentials and magnetic fields were employed. It was found that both local and global discretization influence source reconstruction. Only at a minimum triangle side length of 10 mm was it possible to achieve stable results for MEG and EEG. In order to obtain acceptable errors within the stable region, the ratio of dipole depth to triangle side length must not be less than 0.5. The results obtained from a comparison of the different brain regions indicate that the similarity to spherical geometry might well have an influence on the estimated dipole location, but not so much on its strength. Source reconstruction employing quasispherical correction was found to be the most stable, in particular in the case of coarse BEM discretization. PMID- 9410155 TI - [Evaluation of possible measuring errors from overlaying pressure components in invasive blood pressure recording with external transducers]. AB - Invasive arterial blood pressure measurement using external transducers is a routine measure in intensive care medicine and anaesthesiology. Despite the frequency of its use, measuring errors may occur that can strongly affect the significance of the results. The error with the greatest influence on blood pressure measurement arises from an inadequate dynamic behaviour of the measuring chain, and has been investigated in numerous publications. Measuring errors due to hydrostatic and dynamic pressure have not been considered to date. The present investigation describes an analysis of such measuring errors and possible ways of avoiding them. A side-hole catheter permits pressure measurements to be made without the measuring error caused by dynamic pressure behaviour. However, both analytical and experimental studies revealed that, since the velocities involved are small, this error is negligible in blood pressure measurements, and the increased cost of such catheters is therefore not justified. Measuring error due to hydrostatic pressure can be eliminated by using a zero point compensator. Such a compensator that permits vertical changes in the position of the patient without the need for manual resetting of the zero point has now been developed. However, the clinical test showed that the advantages offered by the zero point compensator again do not justify the increased cost. Furthermore, lateral changes in the patient's position during its use can lead to erroneous measurements. PMID- 9410156 TI - [Ceramic cups for hip endoprostheses. 3: On the problem of osseointegration of monolithic cups]. AB - The combination of ceramic with ceramic offers the option of minimising wear and the need for revision in total hip replacement. At first, uncemented monolithic ceramic sockets were implanted. Some of these did not prove successful. The prerequisites for fixation of an implant by bony integration are discussed i.e. the structural and surface compatibility of an implant. The reasons for the high revision rate associated with monolithic ceramic sockets are discussed. Using monolithic ceramic sockets only contact osteogenesis can in principle be achieved, which does not suffice for good long-term fixation. In many cases, soft tissue can be found at the ceramic socket/bone interface, and the ceramic sockets may as a result migrate and penetrate, creating conditions that elevate the rate of wear to unacceptable levels. PMID- 9410158 TI - Machines for automated evolution experiments in vitro based on the serial transfer concept. AB - Two machine setups for automated evolution experiments in vitro are described. Both machines enable the monitoring of growing populations of RNA or DNA molecules in real time using high-sensitivity glass fiber laser fluorimeters and an automated sample handling facility for volumes in the microliter range. Growth conditions are kept constant by means of the serial-transfer technique, that is, the successive transfer of a small fraction of a growing population into a fresh solution containing no individuals prior to the transfer. The serial transfer technique was modified to work with large populations and constant growth conditions. In the single-channel evolution machine isothermal amplification reactions (Qbeta-system, 3SR, NASBA, SDA) are monitored successively in single test tubes. This machine is particularly well suited for the investigation of optimal adaptation to altered environmental conditions, as is experimentally demonstrated in the evolution of an RNA quasi-species using ribonuclease A as the selection pressure. The new variant of RNA appeared very rapidly (within approximately 80 generations) without stable intermediates, and it was selected by steadily increasing the RNaseA concentration during the serial-transfer experiment. The other machine, which is described in the second part of this article, is a consequent extension of the single-channel machine, and was designed to allow the multichannel detection of up to 960 samples simultaneously. Thus, high-throughput screening can be applied to evolution experiments. In addition to monitoring isothermal amplification reactions, it is also possible to follow PCR amplifications through thin plastic foils. Initial experiments have demonstrated the suitability of the apparatus for uniformly processing samples and for performing thermocycling. PMID- 9410157 TI - [Glutardialdehyde and formaldehyde biosensors]. PMID- 9410159 TI - [A model of regeneration of the liver damaged by dipin]. PMID- 9410160 TI - [Resistance of the terminal polymodal C-units of the cat skin to during mechanical and heat stimulation]. PMID- 9410161 TI - [Mechanism of the vagotropic effect of somatostatin]. PMID- 9410162 TI - [The role of the vagus nerve in the anti-arrhythmic effect of DAGO in acute myocardial ischemia]. PMID- 9410163 TI - [Adaptation to stress in prevention of acute hypotension and hyperactivation of the endothelium in heat shock]. PMID- 9410164 TI - [Does the pleura determine the paradoxical phenomena in the respiratory mechanics]. PMID- 9410165 TI - [Effect of preliminary adaptation to the transauricular electrical stimulation on the catecholamine and met-enkephalin levels in the heart and adrenals of the rats during stress and acute myocardial infarction]. PMID- 9410166 TI - [Hydra peptide morphogen attenuates poststress disorders in the white rats]. PMID- 9410167 TI - [The role of nitric oxide in mechanisms of the nephrotoxic effect of verografin]. PMID- 9410168 TI - [Functional and biochemical correlates of hypoxic shock: cooperative effect of the regulatory peptides]. PMID- 9410169 TI - [Disturbances in the energy metabolism induced by multiple ischemic preconditioning worsen the recovery of the heart function in reperfusion]. PMID- 9410170 TI - [Functional activity of the hepatocytes in the liver fragments in vitro: dependence on the sizes of fragments and duration of their cultivation]. PMID- 9410171 TI - [Effect of the activators on neutrophil motility]. PMID- 9410172 TI - [The role of alpha2-adrenoreceptors in tolerance to complete cerebral ischemia]. PMID- 9410173 TI - [Characteristics of the response of the middle cerebral artery to serotonin]. PMID- 9410174 TI - [Evaluation of the effect of the ointment containing collagenase of the Kamchatka crab on the experimental infected wound]. PMID- 9410175 TI - [Isolation and characteristics of the soluble form of beta-amyloid and apolipoproteins from the cerebrospinal fluid]. PMID- 9410176 TI - [Study of stability of the avirulent phenotype and ability to the limited persistence in the mice of the avirulent mutant of Salmonella enteritidis]. PMID- 9410177 TI - [Immunohistochemical analysis of localization of the somatotropic hormone receptors in the rat liver cells depending on sex and hormonal status]. PMID- 9410178 TI - [Effect of genotype on the hormonal activity of the Leydig cells during stress in inbred strains of mice]. PMID- 9410179 TI - [Conversion of androstenedione in the lymphocytes infiltrating the breast tumor tissue]. PMID- 9410180 TI - [The role of serum humoral factors in metastasis and recurrence of Ehrlich carcinoma in mice]. PMID- 9410181 TI - [The role of enzymuria in evaluation of the nephrotoxicity of antineoplastic chemotherapy in children]. PMID- 9410182 TI - [The experimental study of metabolic correction of hypoxic conditions of the mother, fetus, and the offspring of rats using the new amino acid composition MP 33]. PMID- 9410183 TI - [Tissue and intracellular reorganization of the mouse myocardium induced by the hypogeomagnetic field]. PMID- 9410184 TI - [Comparative aspects of the post-traumatic and prenatal angio- and myogenesis in the mammals]. PMID- 9410185 TI - [Results of transplantation of the nonpreserved autologous and cryopreserved allogenic valve containing segments of the dog vein]. PMID- 9410186 TI - [Dynamics of the reparative and adaptive processes in the myocardium in response to high intensity laser radiation (an experimental morphological study)]. PMID- 9410187 TI - [Autoradiographic analysis of proliferation of the gum epithelium in chronic gingivitis]. PMID- 9410188 TI - [The use of perftoran for the lengthening of the period of tolerance to the experimental acute lethal hypoxia (the apnea model)]. PMID- 9410189 TI - [Correlation between the anatomical characteristics of aorta and atherosclerosis manifestations]. PMID- 9410190 TI - [Effect of asphyxia on the adenylate cyclase activity in the cat cerebral cortex]. PMID- 9410191 TI - [Effect of KLN-93 on ventricular fibrillation induced by reperfusion or electrical stimulation in cats]. PMID- 9410192 TI - [Characteristics of the external respiration as parameter of activity of cell respiration enzymes in the rat brain]. PMID- 9410193 TI - [Anticonvulsant and neurotoxic effects of lamiktal (lamotrigine) in combination with other anticonvulsants]. PMID- 9410194 TI - [Effect of interleukin-1 beta on platelet aggregation in August, Wistar, and VEG rats during acute emotional stress]. PMID- 9410195 TI - [Characteristics of the pressor response induced by adrenaline in arterial hypertension caused by deficiency of exogenous calcium]. PMID- 9410196 TI - [Do endogenous ligands of peripheral mu- and delta-opiate receptors mediate anti arrhythmic and cardioprotective effects of Rhodiola rosea extract?]. PMID- 9410197 TI - [Effect of piracetam administration during lactation on morphometric parameters of neocortex and hippocampus of one month old rats]. PMID- 9410198 TI - [Changes in the parameters of hemostasis and activity of blood lysosome enzymes in allergic necrotizing inflammation during immunosuppression and immunostimulation]. PMID- 9410199 TI - [Involvement of humoral factors in the regulation of hematopoiesis in cytostatic myelosuppressions]. PMID- 9410200 TI - [Effect of low intensity laser radiation on the physical endurance of animals with altered endocrine status]. PMID- 9410201 TI - [Dextrorphan-binding proteins in the hippocampus of audiosensitive genetically epilepsy-prone rats]. PMID- 9410202 TI - [Content of the major heat shock protein HSP70 in rats with hereditary stress induced arterial hypertension]. PMID- 9410203 TI - [Comparative analysis of electrical brain activity after GABA and glutamate administration: is it possible to have correlation between specific neurochemical changes in the brain and in the EEG?]. PMID- 9410204 TI - [Congenital diabetes induced by heminevrin in rats]. PMID- 9410205 TI - [Changes in the production of active oxygen species in neutrophils caused by contrast media]. PMID- 9410206 TI - [Effect of sex hormones and antihormones on the activity of glutathione-dependent redox enzymes of human erythrocytes]. PMID- 9410207 TI - [Synergistic antiherpetic effect of combined administration of para-aminobenzoic acid and modified nucleosides]. PMID- 9410208 TI - [Effects of radiation from video display terminals of personal computers on free radical processes in rats]. PMID- 9410210 TI - [Lipid peroxidation in blood plasma of patients with osteogenic sarcoma]. PMID- 9410209 TI - [Characteristics of Ca(2+)-induced response of erythrocytes in patients with lung cancer during antineoplastic chemotherapy]. PMID- 9410211 TI - [Intensity of lipid peroxidation as an index of inflammation process in the acute phase of chronic bronchitis]. PMID- 9410212 TI - [Dynamics of myocardial function after transmyocardial revascularization with Nd:YAG-laser (clinical study)]. PMID- 9410213 TI - [Hormonal function of the adrenal glands in humans and monkeys during hemoblastoses and aging]. PMID- 9410214 TI - [Free radical status in human blood leukocytes after hyperbaric exposure]. PMID- 9410215 TI - [Contractile activity of smooth muscle of the duodenum during duodenal ulcer and its treatment with metacine and proserine]. PMID- 9410216 TI - [Early bronchoscopic and morphological diagnosis and prognosis in thermoinhalation trauma]. PMID- 9410217 TI - [Morphology of the compensatory hypertrophic kidney in rats at high altitude]. PMID- 9410218 TI - [Effect of fibroblasts, collagen, and laminin on the healing of wounds formed after dissection of split skin flaps in rats]. PMID- 9410219 TI - [Effect of local injection of bone growth regulating factors on the maturation of distraction regenerate]. PMID- 9410220 TI - [A model of tachyarrhythmia in awake animals]. PMID- 9410221 TI - [Mass outbreak of tuberculosis at workplaces]. PMID- 9410222 TI - [Effect of treatment with dexamethasone on cardiovascular responses of adrenergic agents]. AB - Cardiovascular responses to several agents should be modified by glucocorticoid administration in the rat. We investigate the response to adrenergic agonists such as phenylephrine, noradrenaline, clonidine and isoproterenol and ganglionic blocking agent such as hexamethonium in conscious rats treated during 7 days with dexamethasone. Wistar rats were treated with either Dex (150 micrograms daily x 7 days, p.o.) or water. Mean arterial pressure were calculated from the intraarterial recordings of blood pressure. No differences in basal mean arterial pressure were seen between dexamethasone and control groups of rats. Phenylephrine and noradrenaline showed a pressor effect in control rats that was reduced by dexamethasone treatment. Clonidine showed similar pressor effect in both groups of rats but ten minutes after drug administration, a light hypotension was seen in dexamethasone rats. Isoproterenol and hexamethonium showed a similar hypotensive effect on control and dexamethasone rats. In conclusion, dexamethasone treatment should reduce the pressor responses to phenylephrine and noradrenaline. Moreover, the alpha adrenergic agonist clonidine showed a hypotensive effect in dexamethasone treated rats, although the response of isoproterenol and hexamethonium remains unchanged. PMID- 9410223 TI - [Brachytherapy of solid tumors. Use of chromic phosphate colloid]. AB - With the purpose of studying the effectivity of an intratumoral single dose of chromic [32P] phosphate (Phosphocol) for the treatment of solid tumors, studies of bioelimination, biodistribution and therapeutic action were carried out in rats with experimental induced tumors. The results show that the percentage of total elimination is equal to 29.76 +/- 9.60% with a higher percentage in faeces 23.28 +/- 8.81% than in urine 6.48 +/- 2.11%. Biodistribution studies show that, 51.61 +/- 5.82% of the injected activity is found in the tumor while in organs with reticuloendothelial cells, the percentage of activity was 13.09 +/- 5.15% in liver and 2.88 +/- 1.23% in lung. On the other hand, when therapeutic action was evaluated, we found that the percentage of tumor regression (P.T.R) was 61.0% for the injected tumors. It is important to point out that 4 of the treated animals show bioelimination patterns in which the elimination rises suddenly at some time of the study. These results demonstrate that the use of this kind of colloids is not to be recommended for the treatment of solid tumors with moderated degree of vascularization, since its mobilization from the injection point may result in the consequent irradiation of different organs that are not under treatment. PMID- 9410224 TI - [Helicobacter pylori. Current concepts]. PMID- 9410225 TI - Proceedings of the South-Eastern Organ Procurement Foundation Meeting. New Orleans, Louisiana, USA, 30 January 1997. PMID- 9410226 TI - [Incidence of posterior vitreous detachment in the elderly]. AB - Few studies have so far investigated the frequency of posterior vitreous detachment in healthy eyes, and most of these have been biomicroscopic investigations. Since the reported data sometimes vary widely, especially in older age-groups, we conducted sonographic investigations on the frequency of posterior vitreous detachment in old and very old patients. METHOD: A total of 712 eyes in 404 patients aged 65 years and older were studied. Eyes that had previously been operated on, eyes with proliferative retinal disease, and eyes with an ametropia exceeding +/- 3.0 diopters were excluded from the study. The vitreous body was investigated using kinetic B-mode sonography. It was classified as completely attached, completely detached, or partially detached. RESULTS: In the 65- to 69-year-old age-group, only 12 of 105 eyes (11%) exhibited a complete posterior vitreous detachment, while the vitreous body was still completely attached in 75 eyes (71%). A marked decline in the number of eyes with a completely attached vitreous body was first seen in the 80- to 89-year-old age group. Even in this group, however, the vitreous body was still completely attached in 114 of 263 eyes (43%), whereas it was partially detached in 28 eyes (11%). Only in 121 eyes (46%) was the vitreous body completely detached. CONCLUSIONS: The frequency of partial or complete posterior vitreous detachment increases with age. However, posterior vitreous detachment in the elderly is rarer than previously thought. PMID- 9410227 TI - [Fixation behavior in Stargardt disease. Fundus-controlled studies]. AB - Patients with Stargardt's disease often show a significantly reduced ability to read despite fairly good visual acuity. We evaluated whether fundus perimetry with simultaneous observation of the point of fixation can help to explain these difficulties. METHODS: A total of 40 eyes in 21 patients with Stargardt's disease were examined by means of automatic fundus threshold perimetry and special fixation tasks using a scanning laser ophthalmoscope (SLO). RESULTS: During fundus perimetry 19 eyes showed a movement of the mean fixation point towards the top, and five to the right of the central scotoma. At the same time, the variation around the mean fixation point was orientated vertically. We observed an alternation between two different points of fixation in another eight eyes. While one fixation point was usually located in the center inside an area of reduced light sensitivity, the second new locus of fixation was at the top of the scotoma (six of eight eyes). Patients tried to fix small targets inside the scotomatous area. CONCLUSION: Our results demonstrate that patients with Stargardt's disease exhibit a typical pattern of fixation. The development of eccentric fixation occurs in three steps: The initial phase is characterized by central fixation with decreased stability. In the intermediate phase, we observed alternation between central and eccentric fixation. Finally, patients develop a constant eccentric fixation. These results may explain the difficulties in reading which many Stargardt's patients experience and the problems in prescribing corrective devices for these patients. PMID- 9410228 TI - [Metalloproteinase stromelysin. Expression in human retinal pigment epithelium (RPE)]. AB - BACKGROUND: Under normal circumstances the retinal pigment epithelium (RPE) does not undergo cell division. After retinal detachment or the development of choroidal neovascularization (CNV) it can be induced to reenter the cell cycle. The RPE cells proliferate and dedifferentiate. A prerequisite for proliferation of RPE cells is degradation of the extracellular matrix (ECM), which may be induced by metalloproteinases. We investigated the potential role of human RPE cells in the expression of the metalloproteinase stromelysin. MATERIALS AND METHODS: Human RPE cells were cultured from donor eyes. Stromelysin was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) from mRNA. RESULTS: Human RPE cells in culture express stromelysin. Its expression is enhanced by tetraphorbolacetate (TPA). CONCLUSIONS: Stromelysin generally degrades important constituents of the ECM. This may induce the detachment of RPE cells from the basement membrane and initiate RPE proliferation and dedifferentiation. PMID- 9410229 TI - [Retinal detachment in Ehlers-Danlos syndrome. Treatment by pars plana vitrectomy]. AB - Ehlers-Danlos syndrome (EDS) is an hereditary connective tissue disorder caused by defective collagen synthesis, the main features being hyperelasticity and vulnerability of the skin, recurrent bleeding from fragile blood vessels, and secondary deformities of the joints. Ocular involvement is a rare occurrence, e.g., corneal and scleral rupture from minor blunt injury, lens displacement, rhegmatogenous retinal detachment. To date, few reports exist concerning the treatment of retinal detachment in Ehlers-Danlos syndrome, all of them dealing exclusively with conventional scleral buckling surgery. PATIENT AND METHODS: We report on a 47-year-old male patient suffering from EDS type VI (so-called ocular type, lysine-hydroxylase deficiency). He presented with rhegmatogenous retinal detachment in his only eye. A scleral buckling procedure was not feasible because of marked scleral atrophy. A three-port vitrectomy was therefore carried out. RESULTS: During the operation, pronounced choroidal detachment and bleeding developed, subsiding within weeks postoperatively. Closure of the sclerotomies was difficult due to scleral thinning. Two revitrectomies were necessary because anterior PVR with traction retinal detachment occurred. The last revitrectomy was performed 18 months ago, and the retina has been completely reattached under 5000 cs silicone oil since then. Visual acuity is 0.1. CONCLUSION: Primary vitrectomy permits successful treatment of retinal detachment in EDS patients if a buckling procedure cannot be performed because of scleral atrophy. However, serious complications may occur. Surgical procedures other than primary vitrectomy should therefore always be carefully considered, e.g., pneumatic retinopexy, temporary balloon, dura patch with episcleral pocket. PMID- 9410230 TI - [Rhegmatogenous retinal detachments. Seasonal variation and incidence]. AB - Seasonal variations in the relative incidence of rhegmatogenous retinal detachment were reported as a trend in several studies on the database of relatively limited cohort sizes. PATIENTS AND METHODS: An analysis on this topic was performed with a long-term database. A total of 3073 files of patients with rhegmatogenous retinal detachment, covering 11 years of observation, were reviewed for this study. Patients with signs of long-standing detachment or other predisposing diseases were excluded, so that a basic study population of 2314 patients remained for the analysis. RESULTS: The averaged seasonal incidence of rhegmatogenous detachments revealed a significant (P < 0.005) mid-summer peak (n in July = 228) and a winter trough (mean of December-January = 161; difference = 36%). Phase and curve fitting of the seasonal variations in the number of retina detachment cases was similar to the seasonal variation of the astronomic duration of the day (P < 0.001). The relation was closer compared to the average duration of light exposure per day calculated from behavioural data and the astronomic length of light phase (P < 0.0002). CONCLUSION: This long-time study revealed a close correlation of the relative seasonal incidence of retinal detachment and the seasonal variation of light hours per day. So far, this observations cannot be explained pathogenetically. Two basic hypotheses ought to be investigated further: (1) The influence of light on the generation of toxic oxygen radicals and the subsequent destruction of the vitreous and (2) possible light-induced changes in vitreoretinal adhesion. PMID- 9410231 TI - [Complicated cataracts in various forms of retinitis pigmentosa. Type and incidence]. AB - PURPOSE: To study the incidence and types of cataract in retinitis pigmentosa (RP) and their variations among different forms of RP. PATIENTS AND METHODS: This analysis was based on data from 473 patients with RP (autosomal dominant, n = 87; autosomal recessive, n = 79; x chromosomal recessive, n = 23; simplex RP, n = 215; Usher's syndrome n = 80; M. Refsum and others, n = 9) that were retrieved from the literature and patient charts in our clinic. RESULTS: Posterior subcapsular cataract (PSC) developed with the following frequencies for the different genetic types of RP: autosomal dominant, 45.3%; autosomal recessive, 44.0%; x chromosomal recessive, 40.7%; simplex RP, 46.1%; Usher's syndrome, 52.9%. PSC was the only type of lens opacity in patients with Usher's syndrome and autosomal recessive RP.PSC development correlated with early onset of RP symptoms. Nuclear cataracts showed a statistically significant higher frequency in patients with simplex RP (14.8%) than in other genetic types (0-5.9%) (P < 0.01). In addition, nuclear cataracts developed in simplex RP at a significantly later age (69.6 +/- 12.4 years) than PSC (44.4 +/- 12.3 years) (P < 0.001). Patients with cataracts showed significantly worse visual fields than patients with clear lenses (P = 0.00067). CONCLUSIONS: The typical RP cataract (PSC) was found in similar frequencies among all genetic types of RP.PSC was the only type of lens opacity in patients with Usher's syndrome and autosomal recessive RP. Nuclear cataracts developed on average 20 years later than PSC and had their highest incidence in patients with simplex RP. Patients with cataracts showed significantly worse visual field results, indicating a more pronounced retinal pathology. PMID- 9410232 TI - [Cataract surgery in narrow pupil and postoperative fibrin reaction, especially after sphincterectomy]. AB - BACKGROUND: Since increased intraoperative iris irritation can lead to increased postoperative inflammation, we are interested in postoperative reactions to several varied surgical procedures. We performed pupil stretching, iridotomy with iris suture, and partial sphincterectomy. MATERIALS AND METHODS: From January 1995 to January 1996, 100 patients (103 eyes) with narrow pupils underwent cataract surgery. In 13 eyes a iridotomy and iris suture were performed, in 28 eyes a partial sphincterectomy. In 62 eyes there was no surgical intervention after pupil stretching. Postoperative examinations were carried out in the early postoperative phase (up to 5 days postoperative) as well as 4 weeks after surgery. Fibrin reaction was classified according to 3 grades, (1) faint fibrinous threads, (II) fibrinous net, and (III) membranous fibrin exsudation. RESULTS: In the group without additional surgery there was a fibrinous reaction of grade I and II in 11 eyes. This complication occurred after iridotomy and iris suture in four cases and after sphincterectomy in four cases. While the intensity of fibrinous reaction was comparable in the first two groups, the fibrinous reaction after sphincterectomy was more intensive in one eye (grade III). However, response to intensive local antiphlogistic therapy was good. Frequency of fibrinous reaction in the three groups was statistically not significant. It should be pointed out that there was a partial restoration of pupil movements through sphincterectomy: mean pupil diameter was 4 mm, and 2.5 mm under exposure to light. Three patients had no pupillary reaction at all. CONCLUSIONS: Partial sphincterectomy facilities intraoperative manipulations of cataract surgery. Postoperative inflammatory reaction is rare and was very responsive to medical treatment. The reconstruction of pupillary movement is part of full visual function and, last but not least a round pupil is aesthetically more desirable. PMID- 9410233 TI - [Haptic position of iris-fixed posterior chamber lenses. Determination by ultrasound biomicroscopy]. AB - In eyes lacking adequate posterior capsular support, fixation of posterior chamber intraocular lenses (IOL) by iris sutures has become one of the methods available. Ultrasound biomicroscopy (UBM) allows detection of the haptic position postoperatively and determination of its relationship to adjacent intraocular structures. METHODS: Thirteen patients with iris-sutured IOLs were examined pre- and postoperatively with UBM. The examination included locating the haptic and tracing the haptic to the position closest to other uveal structures. RESULTS: The position of all 26 haptics was determined. All haptics were in touch with the iris. In the 12-clock position 11 haptics did not touch other intraocular structures. In one case the haptic was located in the ciliary sulcus, and in another case it touched the ciliary body. In the 6-clock position two haptics were located in the sulcus region, and seven were located posterior to the ciliary processes. Four haptics did not touch any other intraocular structures. CONCLUSIONS: In most cases surgical placement of iris-fixed lenses is a blind procedure. UBM is an adequate method of determining the position of IOL haptics postoperatively. PMID- 9410234 TI - [Peripheral retinal cryocoagulation. Long-term outcome]. AB - Peripheral retinal cryotherapy alone, without additional cyclocryotherapy, can for the short term lead to intraocular pressure reduction and regression of neovascularization. As there is little information concerning the long-term results of this treatment we reexamined all patients treated with peripheral retinal cryotherapy at the Department of Ophthalmology at Freiburg University between 1979 and 1990. PATIENTS AND METHODS: Neovascularization was secondary to proliferative diabetic retinopathy in 33 eyes (65%), secondary to retinal vascular occlusion in 13 eyes (25%), and secondary to a combination of both in 5 eyes (10%). Usually, 2 quadrants of each eye were treated with 9 applications per quadrant. RESULTS: Intraocular pressure was adequately reduced in 49% of the eyes and rubeosis iridis regressed in 57%. Visual acuity deteriorated in 53% of all cases. CONCLUSION: Intraocular pressure and rubeosis iridis can in the long term successfully be treated with peripheral retinal cryotherapy. Despite good intraocular pressure control and regression of rubeosis iridis visual acuity does not improve. PMID- 9410235 TI - [Ocular pulse amplitude and local carbonic anhydrase inhibition]. AB - Ocular perfusion is increasingly being discussed in the pathogenesis of glaucoma. The present study was designed to investigate ocular pulse amplitude (OPA) in primary open-angle glaucoma (POAG) patients with elevated intraocular pressure (HTG) and non-glaucomatous controls (CTL) following topical application of the carbonic anhydrase inhibitor dorzolamide. METHODS: OPA (Ocular Blood Flow System, OBF Labs UK) intraocular pressure (IOP), heart rate (HR), and systolic (BPsyst) and diastolic (BPdiast) bronchial artery pressures were measured before and 2 days after initiating treatment in 33 cataract patients with n = 14) and without (n = 19) POAG. RESULTS: Following application of dorzolamide, IOP (mmHg) in drug treated HTG and CTL eyes was highly significantly reduced (p < 0.001) and in vehicle-treated HTG and CTL eyes significantly reduced (p < 0.03) compared to respective baseline measurements. OPA (mmHg) in drug-treated HTG and CTL eyes was significantly increased (p < 0.05) and in vehicle-treated eyes in affected compared to respective baseline measurements. Systemic perfusion parameters were also unchanged. CONCLUSION: Dorzolamide increased OPA in HTG and CTL eyes. An increase in OPA may improve the prognosis of HTG. PMID- 9410236 TI - [Selective excimer laser ablation of the trabecular meshwork. Clinical results]. AB - PURPOSE: Examination of the efficacy of pore formation in the trabecular meshwork by excimer laser to reduce intraocular pressure in glaucoma eyes. PATIENTS AND METHODS: In 27 consecutive eyes with chronic simple glaucoma and 8 eyes with low tension glaucoma, 3 to 5 pores were ablated into the trabecular meshwork with an excimer laser (308 nm, 35-55 mJ/mm2), creating an open communication between the anterior chamber and Schlemm's canal. This was accomplished by the use of a 400 micron quartz fiber and a modified Trokel goniolens. All patients were candidates for trabeculectomy because visual fields continued to deteriorate in spite of maximum medication. RESULTS: Intraocular pressure was median reduced by 7 mmHg (range 10.5 to 1.5 mmHg) in 22 of 27 eyes with chronic simple glaucoma over a median follow-up of 7 months. In 12 eyes, further medication has to be continued, yet at a lower dose and lower level of intraocular pressure. In five eyes therapy failed. In three of these eyes, a trabeculectomy had to be performed. In eight eyes with low-tension glaucoma, a median reduction of intraocular pressure of 5 mmHg (range 10 to 0.5 mmHg) was accomplished over a median follow-up of 7 months. In five of these eyes, further medication on a lower level was continued. No further surgery was necessary. CONCLUSIONS: With the microsurgical method of pinpoint ablation of the trabecular meshwork by excimer laser, intraocular pressure was reduced in 30 of 35 eyes over a median follow-up period of 7 months. These results encourage us to continue the development of this procedure, perhaps with a microendoscope. The minimal trauma to the eye of this procedure leaves all other options of surgery open. PMID- 9410237 TI - [Effect of a local nonsteroidal anti-inflammatory drug on postoperative irritation after squint surgeries. Prospective randomized double-blind study]. AB - BACKGROUND: Topical application of a nonsteroidal antiphlogistic drug (NSAD) is often recommended to minimize postoperative lid and conjunctival edema. However, the effectiveness of such therapy remains controversial. PATIENTS AND METHODS: The study presented included 100 patients aged 23.6 +/- 19.5 (5 to 77) years, who were randomly assigned to two treatment groups. Group 1 included 50 patients who underwent squint surgery, and who received Flurbiprofen ED twice a day from the first preoperative day to the third postoperative day. Group 2 served as a control group. These patients were treated with the same regimen using artificial tears. The extent of lid and conjunctival edema was assessed and scored from 1 to 4 on the fourth postoperative day. RESULTS: Squint surgery was performed on 129 eyes of 100 patients. In 32 eyes (24.8%) one muscle was operated on. In 79 eyes (61.2%) 2 muscles and in 18 eyes (14%) three muscles were operated on. The average postoperative conjunctival edema score was 2.08 +/- 0.62 (Flurbiprofen group) and 2.04 +/- 0.67 (control group), respectively. The scoring values for lid edema were 1.90 +/- 0.54 and 1.82 +/- 0.43. The differences between the two groups were not statistically significant (P = 0.21 and 0.39; Mann-Whitney test). No influence of the patient's age on the development of lid and conjunctival edema could be detected (P > 0.08). However, the number of muscles that were operated on had a significant influence on both parameters (P = 0.01 and 0.028). Single muscle operations revealed the lowest scaling values (P < 0.02 and P < 0.04). No difference was found whether 2 or 3 muscles were operated on (P = 0.24 and 0.12). CONCLUSIONS: No beneficial effect of topical application of NSAD on postoperative lid and conjunctival edema could be found in this controlled study. PMID- 9410238 TI - [Quantitative and objective follow-up of papilledema with the Heidelberg Retina Tomograph]. AB - The Heidelberg retina tomograph (HRT) is a new instrument to analyze the three dimensional structure of the retina. It is based on confocal laser scanning technology. Scientific interest has so far focused on the follow-up of glaucomatous optic nerve head damage. As parameters such as cup depth and cup volume can be reproduced with a high degree of accuracy, this new instrument may prove to be an excellent tool to follow up swelling of the optic disk. PATIENTS AND METHOD: A total of 21 patients suffering from optic disk swelling of various etiologies were included in a pilot study. Measurements were taken at six different time points. The maximum optic disk elevation and the volume of swelling were evaluated. Visual acuity and the visual field were determined, and fundus photographs were taken. RESULTS: A reduction in optic disk swelling over time may be demonstrated by both morphological parameters and correlates with improvement in fundus changes. The change in the volume of swelling is greater than the change in maximum disk elevation. The course of optic disk swelling differs between patients with anterior ischemic optic neuropathy and those with pseudotumor cerebri. No correlation was found between our measurements with the HRT and functional parameters (visual acuity and visual field). CONCLUSION: The HRT is a good tool for the follow-up of optic disk swelling, particularly optic disk elevation due to increased intracranial pressure. PMID- 9410239 TI - [Asymmetric corneal tunnel incision for routine phacoemulsification and lens implantation]. AB - This study was designed to investigate whether it is possible to improve wound stability and reduce induced astigmatism of a linear corneal tunnel incision for the implantation of foldable intraocular lenses by radializing wound components. MATERIALS AND METHODS: A 4.1-mm linear and a 2.5 x 2.5-mm L-shaped corneal tunnel incision were compared in 60 and 59 patients, respectively. RESULTS: Complications and induced astigmatism (vector analysis) were significantly less in the L-shaped incision group than in the linear incision group (0.62 +/- 0.53 D vs. 0.84 +/- 0.75 D after 3 months). CONCLUSION: An L-shaped incision seems to be superior to the conventional linear type. PMID- 9410240 TI - [Progressive unilateral exophthalmos. Aspergillosis of the orbits]. PMID- 9410241 TI - [Subretinal surgery. With endoscope into the future]. PMID- 9410242 TI - [Diabetic retinopathy in pregnancy. An overview]. PMID- 9410243 TI - Eosinophils in Allergy and Related Diseases. Proceedings of a workshop. Tokyo, Japan, June 21-22, 1996. PMID- 9410244 TI - [Seroprevalence of HCV in the general population of Niger and in patients with chronic liver diseases: comparison of different second generation tests and PCR]. AB - The aim of our study in Niger was to compare the seroprevalence of hepatitis C in a rural "normal" population and in a group of patients presenting at the hospital with signs of chronic liver disease: to estimate this seroprevalence, we used 4 second generation ELISA screening and 3 confirmatory tests (LIA, RIBA and PCR); genotyping was performed on PCR positive sera, using Inno-LIPA HCV. We could not find a statistically significant difference (Fisher's exact test) between the two groups of healthy and sick people (2.5 versus 5.4% for seroprevalence and 2.5 versus 3.2% for viremia). Our study didn't find any relationship between hepatitis C infection, blood transfusion or surgery; other major ways of transmission of hepatitis C have to be considered. The predominant genotype detected was 2a. PMID- 9410245 TI - [Markers of hepatitis C and E virus in Burundi (central Africa)]. AB - The prevalence of antibodies to the hepatitis C virus (HCV) and hepatitis E virus (HEV) was measured in a group of 129 adults (mean +/- SD age 44.7 +/- 13.5 years) from Bujumbura, Burundi, Central Africa. Sera were tested using a second generation ELISA and LIA for antibodies to HCV (antiHCV), and ELISA for antibodies to HEV (antiHEV). The prevalence of antiHCV was 27.1%, very high, in agreement which data from other countries of Central Africa, hyperendemic area for HVC. The prevalence of antiHEV was 4%, much lower than that of antiHAV (97.7%). In addition to the lability of antibodies to HEV, this difference might be explained by the extensive availability of good-quality of drinking-water in the city. The presence of serological markers of HBV and HIV was not associated with that of antiHCV or that of antiHEV. PMID- 9410246 TI - [Re-emergence of Venezuelan equine encephalitis virus in French Guiana. Apropos of 1 confirmed case]. AB - Venezuelan equine encephalitis (VEE) is a mosquito-borne viral disease that occurs in equine species and in man. The strains can be grouped epidemiologically into two major categories: enzootic and epizootic. Enzootic strains cause sporadic human disease and are not associated with disease among equines. These strains are found throughout Florida. Central America, northern South America and Brazil. Epizootic strains are associated with enormous morbidity and mortality in equine species. In man, VEE virus infections are largely asymptomatic and in children and young adults there is an increased risk of encephalitis and dead. We report the first case in French Guiana of Venezuelan equine encephalitis. Clinical examination and biological studies showed encephalitis, interstitial pneumonia and acute liver failure. Despite an adequate symptomatic treatment, the young patient died five days after her admission in multiple organ dysfunction syndrome. Diagnosis is establishing by virologic test: VEE virus is isolated from the blood. These example of re-emerging infectious disease vividly illustrate that we remains vulnerable and emphasizes the need for an active surveillance system. PMID- 9410247 TI - [Molecular typing of Moroccan strains of Mycobacterium tuberculosis]. AB - The insertion sequence IS 6110 was used to differentiate clinical Moroccan isolates of Mycobacterium tuberculosis by using two non radioactive probes. Among 16 strains isolated from patients clinically related, 10 had similar IS 6110 restriction fragment length polymorphism (RFLP) patterns, confirming that they were derived from a common source. Two strains were isolated from the same patient (sputum, lymph node) showed identical profiles hybridized with IS 6110 element. Four sequential strains isolated from the same patients before treatment and after one year had identical IS 6110 RFLP patterns suggesting relapse and not reinfection. Twenty-one strains with identical drug susceptibility showed different IS 6110 RFLP profiles confirming no correlation between antibiotic resistance profiles and IS 6110 RFLP patterns. Since RFLP analysis by using IS 6110 element is a useful tool for the epidemiological survey, of tuberculosis. PMID- 9410248 TI - [Bacterial diversity during the cholera epidemic in Dakar, Senegal (1995-1996)]. AB - Vibrio cholerae O:1, serotype Ogawa and biotype El Tor (76.1%) was responsible of the outbreak of cholera in Dakar, Senegal (1995-1996). However, other bacteria were isolated, particularly Vibrio cholerae non O:1/non O:139, Vibrio fluvialis, Vibrio alginolyticus. Vibrio parahaemolyticus, Salmonella sp.p, Shigella sp.p (23.9%). The Vibrio cholerae O:1 strains are multiresistant to sulfonamide, cotrimoxazole and chloramphenicol. 97% were also resistant to O/129 compound. Fluoroquinolone and 3rd generation cephalosporins were the more efficient antibiotics (100%). PMID- 9410250 TI - [Prevalence of intestinal parasitism in the public laboratories of Martinique: development from 1988 to 1995]. AB - This survey drew up the epidemiological situation of intestinal parasitism in Martinique in 1994-1995. 13,978 stool specimens collected in 1994-1995 were tested by parasitologic examination. Stool specimens were from patients hospitalised in the 3 principal hospitals of Martinique or coming to the Laboratoire departemental d'hygiene. The parasitism rate was 8.73%. This study showed a significant reduction of intestinal parasitism between results of 1988 and results of 1994-1995. The oro-faecal parasitism was not very important that reflected the good economic and sanitation level of Martinique. On the other hand, regarding the important prevalence of parasitism with Strongyloides stercoralis and hookworm, it would be good to improve detection, sanitary education and know better local contamination factors to decrease the prevalence of this parasitism. PMID- 9410249 TI - [The reconquest of the Madagascar highlands by malaria]. AB - A strong malaria epidemic with a high mortality rate occurred on the Madagascar Highlands in 1986-88. Vector control and free access to antimalaria drugs controlled the disease. The authors have searched for the causes of the epidemic to propose a strategy avoiding such events. The Highlands on Madagascar were known as malaria free. In 1878 a very severe epidemic flooded all the country. Development of irrigated ricefields which house both An. arabiensis and An. funestus had created a new anthropic environment. Moreover manpower imported from malarious coastal areas for rice cultivation and also for building large temples, could have brought P. falciparum. After several outbreaks the disease became endemic up to 1949. In 1949 a malaria eradication programme based on DDT spraying and drug chemoprophylaxis and chemotherapy was launched. By 1960 malaria was eliminated and DDT spraying cancelled. Only 3 foci were kept under surveillance with irregular spraying until 1975. The prophylaxis and treatment centres ("centres de nivaquinisation") were kept open up to 1979. The catholic dispensary of Analaroa, 100 km N.E. of Tananarive, opened in 1971 and worked without interruption up to now. The malaria diagnosis has always been controlled by microscopy. Its registers are probably the more reliable source of information on malaria in the area. They show that malaria was already present on the Highlands in 1971 but at a low prevalence; in 1980 when the "centres de nivaquinisation" were closed the number of cases increased by three times the progressive increase of the number of cases became exponential from 1986 to 1988 which was the peak of the epidemic; malaria remained at a high level until the end of 1993; yearly DDT spraying since 1993 have decreased the number of malaria cases among the dispensary attendants by 90%. The epidemic peak of 1988 was well documented by the Pasteur Institute of Madagascar around Tananarive. Before the epidemic started it was observed a come back of An. funestus which had been previously eliminated of most of the villages by DDT spraying. More than an epidemic the malaria increase in 1988 was a reconquest by malaria of the land from which it had been eliminated in the years 1950. This episode became dramatic because the lack of immunity of the population and the shortage of medicaments. The global warming which was advocated to explain the epidemic has no responsibility because the temperature on the Madagascar Highlands has not changed during the last 30 years. Also the cyclones do not seem to have played any role. It is very likely that the gradual decline of control measures, first DDT spraying, later drug distributions, had the main responsibility in the Highlands drama. Everywhere An. funestus reached a high level during the time where the parasite reservoir was rebuilding. They synergised each other. These findings should be taken in account in drawing the strategy planning for the next years. PMID- 9410251 TI - [First Algerian case of human otomyiasis from Chrysomya bezziana]. AB - In Algeria, Human myiasis, essentially ophtalmomyiasis, are known for a long time. Most of cases are due to Oestrus ovis. In this papers the authors report, in a shepherd, the first case of otomyiasis due to Chrysomya bezziana larvae, a species still unknown in North Africa. This observation which indicates the presence of the species in a Northern part of Algeria is also the first report of the insect outside of its endemic traditional area. PMID- 9410252 TI - [Presence of intestinal microsporidia in Tunisia: apropos of 1 case]. PMID- 9410253 TI - [Historical note on the discovery of cutaneous leishmaniasis transmission by Phlebotomus]. AB - In the last edition of Morton's medical Bibliography (1991) the discovery of the transmission of old world cutaneous leishmaniasis by phlebotomes is attributed to english-speaking authors having published between 1924 and 1942. In fact this discovery resulting from researches undertaken since 1904 is due to the team of the Institut Pasteur d'Algerie (E. & E. SERGENT and collaborators) and was published in 1921. We further recall the two meetings of the Societe de pathologie exotique held in Paris on february 8 & 9 1933 during which were exposed the results obtained in Palestine & Syria by ADLER & THEODOR (1925-1929) who implicitly recognized the priority of the French workers of the Institut Pasteur d'Algerie. PMID- 9410254 TI - [Nephropathy and filariasis from Loa loa. Apropos of 1 case of adverse reaction to a dose of ivermectin]. AB - A patient with a filarial infection due to Loa loa, received single oral dose of ivermectin A few hours after, he has presented a severe renal impairment. Native of the Cameroons, he was also treated for malaria. A renal biopsy has been done, because of chronic proteinuria. It has showed microfilariae in glomeruli, thickening of the basement membrane and a segmental and focal mesangial hyalinosis. By immunofluorescence microscopy there were granular deposits of IgM and C3 along this membrane. Electron microscopy has showed subepithelial electron dense deposits. This suggests that the renal disease is immunologically induced. During the therapy, adverse reactions seemed to be related with massive liberation of filarial antigens. This is well known with diethylcarbamazine but also now, sometimes with ivermectin. PMID- 9410255 TI - [African histoplasmosis with ganglionic localisation. Apropos of 1 case in an HIV negative patient]. AB - This study concerns one case of ganglionary African histoplasmosis observed in a pregnant woman. The diagnosis of this histoplasmosis case has been based on histological presentation. This patient has a HIV negative serologic reaction. The histoplasmosis clinical presentation is like tuberculosis, so its diagnosis is difficult. The prevalence of this pathology is unknown in our region but it is increasing since the discovery of AIDS. PMID- 9410256 TI - [Buschke-Loewenstein tumor. Apropos of 2 cases in Cotonou, Benin]. AB - About two cases of Buschke-Loewenstein tumor--one of the penis in man infected with HIV and another of perianal area-, the authors insist on the relative frequency of Buschke-Loewenstein tumor in non-circumcised and homosexual groups. They emphasize the continuous precancerous spectrum of Buschke-Loewenstein tumor. PMID- 9410257 TI - [Bullous toxidermatosis and HIV infection in hospital environment in Lome (Togo)]. AB - A transversal study allowed to analyse the different features of severe bullous toxicodermatosis, and its relations with HIV infection. Since March 1992 to December 1995, 15 patients were hospitalized in Lome Teaching Hospital for severe bullous toxicodermatosis: 16 cases of Lyell's syndrome, 8 cases of Stevens Johnson's syndrome and 1 case of ectodermosis. The principal drug which induced these diseases were mostly dominated by the sulfanilamide (n = 7). Nine (9) of these subjects were infected by the HIV (four of whose patients had Lyell's syndrome). Its noted five cases of death, all in the patients with HIV infection. PMID- 9410258 TI - [Erysipelas of the leg in hospital environment in Lome (Togo)]. AB - A study was conducted during 5 years to determine the epidemiological clinical profile and the evolution of legs erysipelas in patients attending Lome teaching hospital. During this period, 60 subjects (43 females et 17 males) were hospitalized for legs erysipelas. The average age was 40 years old. General facilitating factors were noted in 58% of the patients (n = 35), and local in 50% of cases (n = 30). The first line treatment was in all patients penicillin G. Satisfactory results were observed in 70%. Local complications were noted in 10% (n = 6), and recurrence in 17% (n = 10). The erysipela is common in hospital environment. The precocious penicillinotherapy allowed to have satisfactory results. PMID- 9410259 TI - [Stability and antimicrobial effectiveness of Javel water in a tropical hospital environment]. AB - A sodium hypochlorite solution (NaClO) containing 0.5% w/v of available chlorine (0.5% Cl2 or 5000 ppm) is a good and cheap solution for disinfecting material used to medical care. It is effective against bacteria, fungi and viruses. It is easy to prepare. It can be a good solution for antisepsis and disinfection in dispensaries in developing countries. However, it is not always easy to keep it in adequate conditions. That's the reason why we have evaluated the durability of such a solution of NaClO exposed to disadvantageous factors, specially daylight and air, and its antibacterial effectiveness. In this study, after exposure of NaClO containing 4350 ppm of available chlorine to these adverse conditions, the final concentration of NaClO was 2300 ppm of available chlorine and was found effective for decontamination of medical material. A protocol for conservation and use of a sodium hypochlorite solution (0.5% Cl2) in dispensaries of developing countries is proposed. PMID- 9410260 TI - [Natural vector capacity level of Simulium damnosum s.l. (Diptera: Simuliidae) at the ecology station of Tai (Cote d'Ivoire)]. AB - Several studies have been carried out on the transmission of Onchocerciasis by Simulium damnosum s.l. in the forest zone of Cote d'Ivoire. This study, carried out in 1979-1980 was devoted to determine the risk of onchocerciasis transmission inside and outside the rain forest of Tai (5 degrees 50' N-7 degrees 25' W). We present the vectorial capacity of S. sanctipauli in the region of Tai before massive flow of refugees from areas of Liberia without any control Programme. The results of micromorphological technics for determination of S. damnosum adults, showed that mainly females of S. sanctipauli were present. The studied populations had low parturity rates: 39.2% outside and 30.9% of parous flies inside the rain forest. The parasitic rates (0.4% of infectious females outside and 0.1% inside) and their parasitic loads (15 and 3 infective larvae per 1000 parous female respectively outside and inside the rain forest) were low, consequently their vectorial capacity with Onchocerca volvulus was almost non existent in natural conditions. Before massive flow of refugees including persons carrying microfilariae, there were no problem of onchocerciasis within and outside the rain forest of Tai. However, the massive flow of refugees and the deforestation for growing crops can create situations favourable to the installation of more efficient vectors, increase man/vector contact and contribute to more intense onchocerciasis transmission. The monitoring of onchocerciasis transmission is necessary. PMID- 9410261 TI - [Zoonotic sarcocystosis: from sarcocystic coccidiosis to sarcocystic eosinophilic myositis]. AB - This first deals with the taxinomy and general features of the genus Sarcocystis, which leads to the classification of sarcocystoses and their general epidemiology. Enteral forms of the sarcocystoses, i.e. sarcocystic coccidioses are conjured up and their zoonotic aetiology and their differences with other human coccidioses are emphasized. The main part of the paper is devoted to exenteral sarcocystoses, that can include two forms: acute, seldom recognized, and chronic, including eosinophilic myosites. The author discusses the nature of the parasites involved and the aetiology and epidemiology of the disease. PMID- 9410262 TI - [Improving the supervision of physician-patient relations. Utilization of system's methodology]. AB - PROBLEM ADDRESSED: To train residents adequately, major improvements in the supervision of the doctor-patient relationship were needed. OBJECTIVE OF THE PROGRAM: To improve supervision of residents' building and managing doctor patient relationships. MAIN COMPONENTS OF THE PROGRAM: Using a program based on systems thinking, the teachers at one Quebec university introduced major changes: a reduction in the number of doctors involved in supervision; a common conceptual framework enabling supervisors to more adequately observe, analyse, evaluate, and discuss the doctor-patient relationship; and peer supervision (metasupervision). CONCLUSION: The key to acceptance of these changes was the approach used: consultation, team work, and the development of a new culture. Through this program, supervision of the doctor-patient relationship was improved. PMID- 9410264 TI - [The effect of space flight factors on the intact and regenerating skeletal muscles of the extremities in newts]. AB - The influence of space flight factors on the structure of two skeletal muscles of the hindlimbs was studied in newts. Degenerative-atrophic changes took place in the both muscles of the flight group animals. Comparative-quantitative analysis of ultrastructural changes of the muscle tissue has shown that after space flight, the rate of structural defects of the muscle was twice that in animals of the synchronous and laboratory control groups. But these changes were reversible: within 10 days, the structure of the muscle fibers was practically normalized. The influence of space flight on repair regeneration of the hindlimb skeletal muscle was also studied: the muscle was minced and placed back in its bed 14 days before the flight. Under these conditions, the repair regeneration of the muscle was not suppressed, but a trend towards its delay was found. PMID- 9410263 TI - [The synchronization of the rhythm of protein synthesis in hepatocyte cultures occurs during conditioning of the medium with ganglioside GM1 accumulation in the cells: an immunohistochemical study]. AB - We studied the kinetics of proteins synthesis in the cultures of hepatocytes on collagen-coated slides in medium 199 enriched with 0.2 mg/ml albumin and 0.5 microgram/ml insulin and devoid of bovine serum. Circahoral fluctuations of proteins synthesis intensity were found in the monolayer cultures and sparse cultures in a conditioned medium. No protein synthesis rhythm was expressed in a fresh medium after repeated washing of the cultures. Addition of micromolar concentrations of gangliosides GM1 or GD1a to the medium revealed the rhythm in washed cultures. Addition of GT1b or GM3 to the medium did not synchronize the oscillations: the kinetics of protein synthesis did not differ from that in the control space cultures in a fresh medium without exogenous gangliosides. Accumulation of gangliosides GM1 and GM3 in the hepatocytes in vitro upon conditioning of a serum-free culture medium was shown using the indirect immunocytochemical method. The effect was found in dense monolayer cultures and in cultures with separated cells in a conditioned medium. The protein synthesis rhythm was found in such cultures. Gangliosides are weakly expressed in most cells of a repeatedly washed 24-hour monolayer and washed sparse cultures. No protein synthesis rhythm was found in washed cultures. Similar changes in the dynamics of the culture medium conditioning, accumulation of gangliosides in cells and rhythmic activity of cells population confirm the concept of the synchronizing role of gangliosides. PMID- 9410265 TI - [A computer study of the relationship between the value and type of their embryotoxicity and the structure of synthetic analogs of biogenic amines]. AB - We calculated 219 topological, electronic, and steric indices for each of 59 studied embryotoxic benzylalkylamines and indolamines. Statistically significant equations correlating embryotoxicity and five calculated parameters were obtained using the method of step multiple regression. The prognostic power of the equation was 88-90%. Discriminant functions obtained by step discriminant analysis allowed prediction of the superactivity of benzylalkylamines and indolamines with 83-87% probability. PMID- 9410266 TI - [A vitamin-antioxidant diet decreases the level of chromosomal damages and the frequency of gene mutations in irradiated mice]. AB - Two different methods were used to study the effect of the antioxidant vitamins mixture (AVM) containing beta-carotene, alpha-tocopherol, ascorbic acid, rutin, and microelements zinc and selenium on mouse resistance to acute and chronic irradiation. Micronucleus test demonstrated that a daily dietary supplement of AVM reliably decreased the rate of chromosomal damage, namely, X-ray-induced micronuclei in the bone marrow polychromatophilic erythrocytes of aging mice, but did not induce micronuclei formation in the young mice. Assay for somatic mutations in the hypoxanthine-guanine phosphoribosyl transferase (HPRT) locus showed that AVM significantly decreased the rate of genic mutations in mouse splenocytes after chronic irradiation. PMID- 9410267 TI - [The efficiency of the calcitonin-transporting activity of the hemato-C-cellular system in the thyroid gland]. AB - The efficiency of hemato-C-cellular transport of calcitonin into the systemic blood flow was evaluated by the methods of statistical modelling. The linear regression model obtained on 36 rats suggests essential predetermination of the serum concentrations of calcitonin (CT) by the intensity of incretory activity of the thyroid parafollicular epithelium: CT = 13.733+ (0.318 Nvsg; rxy = 0.830; p < 0.001 (Nvsg is numerical density of the sublemmal granules on the C-cells vascular pole). At the same time, the resulting index (CT) depends not only on Nvsg. This index is also influenced by the factors not controlled by the model, such as Hemato-C-cellular delay of calcitonin. These factors determine each certain proportion of the error of prognostication of the calcitonin serum content according to Nvsg. A covariation model CT = bNvsg was plotted in order to determine the total influence of uncontrolled factors. Its error of prognostication was subject to analysis of variance and a confidence interval of biological accessibility of endogenous calcitonin was calculated (73% < FCT < or = 100%; p > or = 0.95). The results obtained suggest the absence of essential intrathyroid loss of calcitonin at the transinterstitial and transmural stages of hemato-C-cellular hormone transfer. PMID- 9410268 TI - [Where to search for the magnetoreceptor]. PMID- 9410269 TI - [The role of osteoclasts in maintaining calcium homeostasis in newts]. AB - The intensity of osteoclastic resorption and calcium contact were investigated in intact limb bones of the newts flown onboard of biosatellite Cosmos-2229 after amputation of their forelimbs and tail. X-ray microanalysis has shown an increase of calcium content in the bones on the 20th day after operation. Histological study revealed activated osteoclastic resorption on the inner surface of long bones. The newts exposed to weightlessness after the operation had the same level of bone mineralization as the operated ground control ones, but the number of polynuclear osteoclasts increased to a lesser extent. PMID- 9410270 TI - [The role of gravitational force in the evolution of living systems (the biomechanical and energy aspects)]. AB - Possible pathways of the origin and development of adaptive mechanisms in the living systems of various levels of organization are considered from the positions of biomechanics and bioenergetics. Main attention is paid to the specific feature of functional rearrangements in the living organisms during change of habitat, at the stage of their exit from water to land. The following problems of interaction between the living systems and gravity during evolution are discussed: cellular level of organization and transition to multicellularity, formation and improvement of the skeletal and skeletal-motor systems in plants and animals, and thermal homeostasis of the living organisms in the gravity field. We showed a leading role of gravity in organization of the morphofunctional status of the living organisms during their evolution. PMID- 9410271 TI - [The conservation of the genetic resources of deer (Cervidae) by the cryopreservation of their germ cells]. AB - Necessity of organization of a cryobank for the sperm of some taxa has been substantiated on the basis of the published data and authors' results concerning the state of genetic resources of the family Cervidae in Russia. A task has also been set to elucidate some factors that may affect the success of cryoconservation, such as the age of male donors; season; duration of sperm storage before and after freezing, including long-term storage; and the possibility of obtaining of sperm from perished males. Results are provided on the influence of these factors of the quality of spermatozoa obtained on three species of the Cervidae. PMID- 9410272 TI - [The caudal gland in the cat is a hepatoid gland]. AB - The cat supracaudal gland is usually considered as a conglomeration of massive sebaceous glands, although the author of the only detailed description of its structure stated that it is a hepatoid gland (HG) of lower order. Investigation of the supracaudal gland in adult cats of both sexes showed that it is a massive layer of HGs opening either into dilated follicles of coarse hairs or into large cisterns. These glands possess all features specific for HGs: polyhedral cells, a network of intracellular canaliculi, abundant cytoplasmic protein granules, release of protein structures from the cells to adjacent canaliculi. The reasons for misinterpreting certain structural properties are discussed. It was shown that the cat caudal gland is a typical "embryonal" HG previously described in dogs and certain hollow-horned ruminants. They secrete protein and hydrophobic lipids, have wide excretory ducts, are connected with coarse hairs, are devoid of cysts, and have no sexual dimorphism. Formation of cisterns, special secretion receptacles of the cat supracaudal gland, was monitored. It begins with stagnation of the lipid-protein secretion in the hair follicle and progresses until transformation of the whole glandular lobe to the cistern and dissolving of the hair shaft in it. PMID- 9410273 TI - [The late sequelae of the action of irradiation during the cleanup of the aftermath of the Chernobyl catastrophe on the concentration of macro- and microelements in human blood serum]. AB - The content of Cu, Fe, Zn, Mg, P, and S in the blood serum and its ultrafiltrate was studied by the inductively coupled argon plasma atomic emission spectrometry in eliminators of the accident at the Chernobyl Nuclear Power Station who worked in the accident zone in 1986-1987. Changes in the mineral equilibrium of the blood serum was expressed in elevated concentrations of the ultrafiltered Cu and S, increased Mg content in the serum, and decreased content of ultrafiltered Zn. The Fe concentration in the serum and ultrafiltrate did not differ from the control indices, thus suggesting balanced Fe transport, accumulation, and mobilization at a remote time after the radiation injury. PMID- 9410275 TI - [The characteristics of the morphological study of the animal population on zonal transects (the initial concepts and assumptions)]. AB - The article briefly describes the original concepts of morphological studies of animal communities. The use of geographic and structural approaches for studying the territorial heterogeneity of communities is considered. The problems of typological continuum of animal communities, principles of their classification and revealing of the environmental factors related to spatial differences of communities are discussed. PMID- 9410274 TI - [Changes in the contractile and bioelectrical activities of the rat small intestine under increased intra-abdominal pressure]. AB - A relationship was established between the increased intra-abdominal pressure and decreased electrical and mechanical motor activity of the small intestine in rats. Possible pathophysiological mechanisms are discussed. A model was developed for estimation of the electrical and motor activity of the small intestine in rats, which can be used for experimental studies of conditions where intra abdominal pressures can be elevated. PMID- 9410276 TI - [Nitric oxide as a factor in the antihypoxic effect of adaptation to physical loading]. AB - It is known that adaptation to exercise enhances the organism resistance to acute hypoxia. However the mechanism of this cross protective effect have been insufficiently studied. The analysis of literature suggests that NO may play a role in the development of the antihypoxic effect of adaptation to exercise. The aim of the present study was to test this hypothesis: first, by evaluating the influence of NO donor and NO-synthase inhibitor on the antihypoxic effect of adaptation to exercise and, second, by evaluating the changes of NO production in acute hypoxia and after a course of adaptation to exercise. It was shown that the NO donor could both reproduce and considerably (three times) potentiate the antihypoxic effect of adaptation to exercise. At the same time, the NO-synthase inhibitor completely suppressed the development of the protective antihypoxic effect of adaptation. After adaptation to exercise, the cerebral NO production was unchanged, while the hepatic NO production doubled. Acute hypoxia induced a biphase change in tissue NO production; initial increase (twofold in the brain) preceded a decrease (by 25% in liver and 37% in the brain as compared to the control). Therefore, the increased NO production following adaptation to exercise can underlie the antihypoxic effect of such an adaptation. PMID- 9410277 TI - Proceedings of the Hematopoietic Stem Cells International Symposium and Workshop. Germany, July 1996. PMID- 9410278 TI - [From spirometry to evaluation of breathing regulation. Issue in honor of Antoni Koziorowski]. PMID- 9410279 TI - [Clinical principles of breathing regulation assessment]. PMID- 9410280 TI - [Chemical control of breathing in patients with obstructive sleep apnea]. AB - The authors have studied chemical control of breathing in 37 normocapnic patients with OSA. These patients had increased apnea-hypopnea index (AHI = 51 +/- 22), obesity (BMI = 32.4 +/- 5.6 kg/m2) and normal lung function tests. Control group consisted of 20 healthy subjects with normal weight (BMI = 23.1 +/- 2.4 kg/m2). Respiratory responses (ventilatory and P0.1) to hypercapnic and hypoxic stimulation during rebreathing tests were measured with computerized methods. The obtained results in OSA patients were compared with the data of the control group. The results exceeding mean values of the control group above 1.64 SD were recognized as hyperreactive responses. The majority e.g. 26 patients (OSA-N) had normal respiratory responses during hypercapnic stimulation. delta V/delta PCO2 = 16.8 +/- 4.5 L/min/kPa, P0.1/delta PCO2 = 3.5 +/- 2.4 cm H2O/kPa/. In remaining 11 patients (OSA-H) respiratory responses were significantly increased delta V/delta PCO2 = 39.1 +/- 18.8 L/min/kPa, P0.1/delta PCO2 = 8.6 +/- 3.9 cm H20/kPa). During isocapnic hypoxic stimulation majority e.g. 25 patients (OSA-H) had significantly increased respiratory responses delta V/delta SaO2 = 3.28 +/- 1.63 L/min/%, delta P0.1/delta SaO2 = 0.54 +/- 0.43 cm H2O/%/. In remaining 12 patients (OSA-N) respiratory responses were within normal limits delta V/SaO2 = 1.2 +/- 0.28 L/min/%, delta P0.1/ delta SaO2 = 0.21 +/- 0.07 cm H2O/%/. The above results indicated, that majority OSA patients (67.5%) had increased ventilatory and P0.1 responses to hypoxic stimulation. Among them also 11 patients had increased respiratory responses to hypercapnia. It seems, that increased respiratory responses to hypoxic stimulus in OSA patients are symptoms of protective reaction to hypoxaemia occurring during repetitive sleep apnoea and reveals increased neuro-muscular output. PMID- 9410281 TI - [Pattern of breathing and obstruction pressure in patients with airway obstruction after inhalation of bronchodilators]. AB - The neuromuscular drive is increased in patients with an airway obstruction. The aim of the study was to estimate an influence of beta-agonist on breathing pattern and mouth occlusion pressure (P0.1) in patients with reversible and nonreversible airway obstruction. Ventilatory function tests, pattern of breathing analysis, mouth occlusion pressure (P0.1) and inspiratory impedance (P0.1/Vt/Ti) were measured in 23 obstructive patients and 20 healthy subjects. In all patients these measurements were repeated 20 minutes after bronchodilator inhalation (0.2 mg fenoterol). During quiet room-air breathing in patients we observed increased Vt, Vt/Ti comparing with healthy persons. The time of inspiration (Ti) and total time (Ttot) were shortened in comparison to our control group. P0.1 and inspiratory impedance were significantly increased (P0.1 3.6 +/- 1.6 vs 1.6 +/- 0.3 cm H2O, p < 0.01 and P0.1/Vt/Ti 6.6 +/- 2.3 vs 3.8 +/- 1.0 cm H2O/L/s). Measurements performed after bronchodilator inhalation revealed decrease of P0.1 and P0.1/Vt/ Ti in patients responsive to beta-agonist (delta FEVI > 15%). In unresponsive patients (delta FEVI < or = 15%) such decrease in neuromuscular respiratory drive was not observed. We conclude that diminishing of increased neuromuscular respiratory drive in patients with reversible obstruction is a consequence of airway resistance decreasing. It seems to be an additional, advantageous for a patient, effect of bronchodilator inhalation. PMID- 9410282 TI - [Clinical use of respiration biomechanics measurements in newborns]. AB - The aim of the study was to assess the clinical usefulness of lung mechanics measurement in newborns with respiratory distress syndrome. Total respiratory system compliance (Crs) by occlusion technique has been measured in 49 pre-term and in 13 fullterm newborns. All measurements were performed during quiet sleep, newborns were shortly disconnected from respirator and breathed through Fleisch "0". During measurements heart rate and saturation were continuously monitored. Airway pressure was measured downstream of pneumotachometer, occlusions were manually performed. Significantly lower mean Crs values were found in newborns treated with IPPV and IMV in comparison with spontaneously breathing extubated newborns and treated by CPAP. Significantly lower Crs values were found in newborns requiring higher oxygen concentrations in comparison with low oxygen dependent. The results show that newborns with respiratory distress syndrome may be clinically well characterised by Crs value. PMID- 9410283 TI - [Is it possible to predict the result of the bronchial provocation test in children with asthma?]. AB - Bronchial hyperreactivity (BHR) to different allergic and non-allergic stimuli is characteristic feature of asthma. Sometimes however it is not possible to perform bronchial provocation test (BPT) assessing reactivity. It was interesting for us if the result of BPT can be predicted on the base of routine lung tests. The aim of the study is evaluation of the relationship between BPT results and baseline lung function tests assessing small bronchi obstruction in children suffering from asthma. Investigated group comprised 139 children aged 7 to 17 years, with episodic, mild or moderate asthma. During bronchial challenge lung function was assessed on the base of spirography and maximal flows at 50% and 25% of forced vital capacity (MEF50 and MEF25) and other indices as surface under end-half of flow-volume curve and mean flow times T50 and T25. The study results confirmed good correlation between BPT result and baseline lung function. Those children which had worse initial lung tests had more pronounced bronchial hyperreactivity. This relationship was the closest in the group of children with small bronchi obstruction. Analysis of correlation showed highly significant relationship between baseline lung function tests and degree of bronchial reactivity. The highest significance was observed for MEF50 and MEF25. We conclude that small bronchi test disturbances in children with asthma could predict with high probability results of bronchial challenge. PMID- 9410285 TI - [Results of respiratory muscle training in patients with chronic obstructive lung diseases with a moderately severe course]. AB - The aim of the study was to testify the effectiveness and acceptance of combination of the resistive inspiratory muscle training and the following walking training in 26 stable COPD pts with FEVI%VC about 50%. Measurements of inspiratory (MIP cm H20) and expiratory (MEP cm H20) muscle strength and also spirometric examinations two times before the training and one time after every month of observation were done. During first 3 months of the training inspiratory resistor (Pflex) was used. Everyday 20 s MVV with smaller orifice of Pflex (strength training) and 30 min quiet breathing by medium orifice of Pflex 3 times a week (endurance training) were performed. In first 3 months of experiment 13 patients resigned of it. Other 13 pts (age 64 +/- 11.9. F-4 with medium FEVI%VC = 47.3 +/- 14.5, MIP = -54.1 +/- 11.4, MEP = = 99.8 +/- 40.5) were trained during 3 months. In 11 of them (FEVI%VC = 50.0 +/- 17.3) MIP improved by 33%. Values of other indices did not change at all. After 3 month of resistive training 11 pts (3-F) were qualified to walking training but only 5 (1-F) of them had finished it. After 5 month of exercises their MIP improved by 74% (p < 0.5) and MEP by 50% (p < 0.05). We conclude that our method of resistive breathing with Pflex is effective and well tolerated by stable COPD pts but accepted only by 50% of them. It may be used as the first step before conditioning training. Inspiratory muscle training is not alternative to pharmacological treatment but is valuable supplementation of it. PMID- 9410284 TI - [Evaluation of respiratory muscle strength in patients with interstitial lung changes based on simultaneous measurement of esophageal and mouth pressure]. AB - A well know good relation between nasal and oesophageal inspiratory pressures exists in healthy and in COPD patients "sniff manoeuvres. Similar results are obtained using "gasp" maneuvers. The aim of the study was to appreciate the usefulness of "gasp" for evaluation of inspiratory muscles strength in ILD patients. 18 ILD patients were examined: group A consisted of 9 pts (8M+IF) aged 35 +/- 8.6 yrs, with static compliance > 70% pred. (mean 98.6 +/- 16.3), group B consisted of 9 pts (6M + 3F) (aged 52 +/- 13.0) with static compliance < 70% pred. (mean 37.2 +/- 12.0), Pmo and Poe (Milic-Emili method) were measured simultaneously during breathing with Pflex device (1.7 mm diameter). Results were stored in a computer for further analysis. In all patients spirometry, plethysmography and maximal inspiratory (MIP) and maximal expiratory (MEP) pressure measurements were performed. Poe and Pmo in group A were nearly the same (8.16 +/- 1.82 vs. 8.35 +/- 2.74 kPa), but in the group B Pmo was lower than Poe (4.81 +/- 1.59 vs. 6.19 +/- 2.03 kPa; p < 0.0005). We conclude that "gasp" - Pmo is a useful method for inspiratory muscle strength measurement only in ILD patients with normal static compliance but in ILD patients with decreased compliance "gasp" - Poe measurement in necessary. PMID- 9410287 TI - [Diaphragmatic electromyogram and respiratory pattern after unilateral and bilateral partial denervation of the diaphragm in the cat]. AB - In the present study we investigated the mechanism of early respiratory compensation of partial paralysis of the sternal and lateral diaphragm due to an unilateral or bilateral section of the C5 rootlet of the phrenic nerves in anesthetized cats. Compensatory effects were evaluated from the recordings of the bilateral diaphragmatic EMGs, neural respiratory pattern and ventilation. The results of the study demonstrate that successive C5 denervation of the diaphragm caused a decrease in the ipsilateral diaphragmatic EMG. Bilateral C5 section evoked an up to 10 percent decrease in minute ventilation. The compensation of the unilateral and then bilateral partial impairment of the muscle function was achieved always by an increase in the neuromuscular projection to the currently contralateral diaphragm. Neural mechanisms of compensation involve a general increase in the respiratory drive, expressed mostly as an increase in the frequency of breathing. The contribution of afferent respiratory muscles to these mechanisms is likely. PMID- 9410286 TI - [Behavior of arterial blood oxygen saturation at night in patients with obstructive lung diseases qualifying for home oxygen therapy]. AB - In COPD patients undergoing LTOT recent ATS and ERS guidelines suggest increase of oxygen flow by I L/min. to avoid severe desaturations during sleep. The aim of this study was to investigate frequency of overnight desaturations in COPD patients qualified for LTOT. We studied 101 consecutive COPD patients qualified for LTOT. Their functional characteristics were as follows: FVC = 2.24 +/- 0.78 L, FEVI = 0.88 +/- 0.39 L, PaO2 = 50 +/- 5 mmHg, PaCO2 = 48 +/- 8 mmHg. Overnight pulse oximetry was performed twice, while breathing air and on supplemental oxygen assuring satisfactory oxygenation (PaO2/O2 at rest when awake = 65 +/- 7 mmHg). We distinguished three groups of patients according to mean overnight arterial blood saturation breathing supplemental oxygen (mean SaO2/O2). First group - 40 patients (39.6%) had mean SaO2/O2 > 93% and time spent in saturation below 90% (T90/O2) = 4.5 +/- 6.7% of the recording time. Second group - 30 patients (29.7%) had mean SaO2/O2 between 90% and 92% and T90/O2 = 32.7 +/- 15.3%. Third group - 31 patients (30.6%) had mean SaO2/O2 < 90% and T90/O2 = 81.8 +/- 16.2% of the recording time. Comparison of ventilatory variables and daytime blood gases in these groups revealed statistically significant elevation of PaCO2 in group 3 (54 +/- 9 mmHg) compared to group 1 and 2 (45 +/- 6 mmHg and 47 +/- 7 mmHg respectively). FVCI, PaO2 and age were similar in all groups. We conclude that around 1/3 of COPD patients qualified for LTOT need increased oxygen flow during sleep. Such need should to be confirmed by the overnight pulse oximetry. PMID- 9410288 TI - [Twenty year observation of pulmonary hypertension in a patient with COPD]. AB - Evolution of pulmonary hypertension over 20 years of observation in a patient with COPD is presented. Originally patient presented with normal pulmonary arterial pressure. First bout of pulmonary hypertension was observed during severe pulmonary infection resulting in respiratory failure. Smoking cessation, regular use of bronchodilators and antibiotics during bacterial exacerbations produced long-term stabilisation of FEV1, blood gases and pulmonary arterial pressure. PMID- 9410289 TI - [The effect of obstructive sleep apnea on pulmonary circulation]. PMID- 9410291 TI - [Use of a nonspecific bronchial provocation test for pre-employment qualification]. PMID- 9410290 TI - [Peripheral arterial chemoreceptor function and mechanisms]. PMID- 9410292 TI - [Inspiratory muscle fatigue as a cause of hypercapnic respiratory failure]. PMID- 9410293 TI - [Respiratory regulation--opioids as factors modifying respiratory drive]. PMID- 9410294 TI - [Hydrogen peroxide in expired breath as a marker of respiratory tract inflammation]. PMID- 9410295 TI - [Evaluation of quality of life and its role in rehabilitation in patients with chronic respiratory diseases]. PMID- 9410296 TI - [Know the age of your lungs]. PMID- 9410297 TI - [Post exertion bronchospasm--usefulness of different methods for evaluating bronchial constriction after exertional provocation]. AB - 55 children with bronchial asthma were tested using three exercise provocation tests: a treadmill, stairways running and cycloergometer. The results were evaluated on the base of the lung function tests, auscultation and airways resistance measurement with occlusion method. Of these three tests the treadmill test seemed to be the most useful to prove bronchial hyperreactivity towards exercise. Stairways running occurred to be very congenial. The study also proved usefulness of auscultation in evaluating of bronchial constriction after exercise. PMID- 9410298 TI - [Children with wheezing as a group with risk of asthma occurrence]. AB - To determine a risk of bronchial asthma appearance in children with wheezing during the first three years, 53 children were examined. They were compared to a group of randomized school children N = 38, and a group of asthmatics N = 42. Among children with wheezing a risk of asthma appearance was 33.8%. In lung function tests no similarities were detected. PMID- 9410299 TI - [Comparison of the effect of two budesonide preparations for treatment of bronchial asthma in children]. AB - In a group of 55 children in the age of 7-14 years with mild or severe asthma, the evaluation of inhaled budesonide (in pulmicort and horacort) effect on bronchial reactivity and adrenal cortex function was made. Budesonide in dose 640 mg or 800 mg daily was administered during 6 weeks. In addition fenoterol was administered in dose 2 x 200 mg daily. Lung function tests, a histamine provocation and cortisol level in serum were done before, after 2 weeks and after 6 weeks in the course of treatment. Effective control upon the disease was observed in 67.9% and 70.4% patients. The result of the treatment was significant decrease in bronchial reactivity toward a histamine. Decrease of FEV1-32% and 34% in the first study was improved after two weeks up to -24% and -17% and after six weeks it achieved-12%. The level of endogenous cortisol in serum didn't show significant decrease in the course of therapy. The ratio of benefits and risk of side effects in the course of budesonide in medium dose during six weeks indicates clearly the first ones. PMID- 9410300 TI - [Prevalence of nasal sinusitis in children with bronchial asthma]. AB - The analysis of prevalence of nasal sinusitis in 558 children 3-6 years old with various pulmonary diseases was made. In a group of 142 asthmatic children 53 (37.3%) had some alterations in sinuses. In another group of children (n = 91) with other atopic diseases, nasal sinusitis were observed in 18 (19.8%) individuals, but in a group of the children with other non-atopic pulmonary diseases (n = 325), nasal sinusitis was found in only 19 (5.9%) individuals. The correlation between the severity of asthma and prevalence of nasal sinusitis was found. In children with severe asthma the abnormality of sinuses were found in over 76% individuals. CONCLUSION: among young children with bronchial asthma nasal sinusitis can be found in 37.3% individuals and it is far more frequent than among patients with other non-atopic pulmonary diseases (5.9%). PMID- 9410301 TI - [Evaluation of the effectiveness and safety of salmeterol in children with nocturnal dyspnea]. AB - 10 children 8-16 years old (mean = 12.9) received at least 4 inhalations of short acting beta-2-mimetics and up to 100 mcg inhaled corticosteroids or/and 5 mg oral prednisone daily. Lung function test values indicated mild/severe asthma in these patients (PEFF 50-80%, FEV1 50-80%, FEV1/VC less or equal 60%). Reversibility of obstruction was at least 15% comparing to the initial values. After two weeks of this treatment resolution of dyspnoea in the night-time and significant decrease of dyspnoea in the day-time were observed. The best bronchodilatation effect was noticed in the third and fourth week of therapy. After discontinuation of Serevent significantly milder course of dyspnoea was observed. PMID- 9410302 TI - [In search of the genotype for atopy]. PMID- 9410303 TI - [The role of food allergy in asthmatic children]. PMID- 9410304 TI - [Antiinflammatory action of corticosteroids in bronchial asthma--mechanism of action and pathophysiologic aspects]. PMID- 9410305 TI - [Stability of inhaled drugs as a requisite for safe therapy as for example, with nebulised glucocorticosteroids]. AB - In inhalation therapy preservation of the stability of an aerosolized drug molecule while nebulisation is an important factor determining its clinical efficacy and safety. The influence of nebulisation with employment of ultrasonic and jet methods using compressed air and oxygen as aerosol carriers on the stability of inhaled hydrocortisone hemisuccinate, budesonide and flunisolide was investigated. Collected aerosol samples were analyzed using thin layer chromatography and compared with standard solution samples in UV-wavelength chi = 254 nm. The appearance of additional fluorescence exctinction points by ultrasonic and air jet methods produced hydrocortisone hemisuccinate aerosol samples and by an ultrasonic method produced budesonide aerosol sample were observed. This could provide evidence for affecting stability and appearance of desintegration products of hydrocortisone hemisuccinate and budesonide while nebulisation using aforementioned methods. In the context of the obtained results the stability of a flunisolide molecule remains unaffected. Furthermore, the process of steroids solution condensation on the membrane of the nebuliser and in inhalation vessel were seen. The phenomena of molecule desintegration and solution condensation as well stated while nebulisation lead to reduction of the inhaled steroid dose accessible for a patient. PMID- 9410306 TI - [Role of theophylline in treatment of chronic obstructive pulmonary disease]. PMID- 9410307 TI - [Anti-inflammatory and immunomodulating action of theophylline]. PMID- 9410308 TI - [Contemporary opinions on the use of new generation H1 receptor blockers for allergies]. PMID- 9410309 TI - [Selected psychological factors and methods of their evaluation in diagnosis and treatment of patients with asthma]. PMID- 9410310 TI - [Education of a patient with asthma]. PMID- 9410311 TI - [Differences in teaching management--education of children with asthma]. PMID- 9410312 TI - Conference on Emerging Diseases. Proceedings. The United States-Japan Cooperative Medical Sciences Program. Tokyo. PMID- 9410313 TI - [Cowper's gland: anatomic, physiological and pathological aspects]. AB - Cowper's glands are small appendages of the male genital tract, that are sometimes observed on intravenous urography voiding films. They are involved in the immune defence of the genitourinary tract, play a role in fertility, and secrete many glycoproteins, including PSA. They can be visualized in the form of a duct image parallel to the urethra, sometimes associated with opacification of the gland on IVU. This image can be differentiated from a fistula, extravasation of contrast agent, urethral duplication, or an artefactual image, by the course parallel to the urethra, the upper limit not exceeding the urogenital diaphragm, and the position of the orifice. These glands can be affected by neoplastic, infectious, stone and especially cystic disease: syringocele. The physician should think of these glands in patients with unusual genitourinary symptoms in order not to miss disease of these organs, and to prescribe appropriate treatment. PMID- 9410314 TI - [Anatomical and clinical effects of neoadjuvant hormonal treatment before radical prostatectomy]. AB - Neoadjuvant endocrine treatment before radical prostatectomy is designed to reduce the positive margin rate and therefore the risk of recurrence. It usually consists of complete androgen blockade for 3 months, but no consensus has been reached concerning the type of blockade and the optimal duration. The main histological changes consist of glandular atrophy with pycnosis and vacuolization of tumour cells. These changes could reflect increased apoptosis. The residual tumour is usually small. PIN become rare, raising the problem of possible chemoprevention. The overall result is a markedly increased frequency of intracapsular tumours and a reduction of at least 50% of positive margins. However, some authors consider that this is simply due to a more detailed histological examination. The frequency of seminal vesicle and lymph node involvement does not appear to be modified. Endocrine therapy could act on the apoptosis-proliferation balance and also decrease the number of circulating cells. However, the intermediate results of randomized studies are contradictory. As improvement in overall survival remains the main objective, only a longer follow-up will be able to clearly determine the value or uselessness of neoadjuvant endocrine treatment before radical prostatectomy. PMID- 9410315 TI - [Evaluation of clinical competence in urology: innovative approach based on performance observation]. AB - OBJECTIVE: The authors present a pilot project for the evaluation of clinical skills in urology using a method of evaluation based on observation of real performance. MATERIAL AND METHODS: An objective and structured clinical examination (OSCE) applied to urology was developed according to a precise predetermined design: 1) Identification of the objectives to be evaluated. 2) Choice of sampling of clinical situations representative of routine urological practice. 3) Construction, on the basis of these cases, of physician-patient interaction stations and question stations, with, for each case, weighting of objectives, construction of observation grids and writing of instructions for candidates, simulated patients and observers. RESULTS: An OSCE circuit of 10 clinical cases and 16 stations was constructed. The main poles of activity and urological settings were represented. Objective complementary investigations, diagnosis and treatment received the highest weightings. The reliability coefficient, the content validity and the construct validity will be verified on the basis of the overall score obtained by candidate. CONCLUSION: Establishment of the score and the expected results in terms of reliability, validity, and feasibility are discussed. The psychometric qualities of the OSCE have been demonstrated. Although a single instrument is not sufficient, it is currently the measuring tool which most closely approximates ideal evaluation of clinical skills. If the feasibility of this method is confirmed, this pilot project in urology could provide a new approach to evaluation strategies and could participate in the current reflection concerning urology training. PMID- 9410316 TI - [Urinary fistula after transplantation: eleven cases]. AB - OBJECTIVES: To specify the anatomical features of urinary fistulas and to evaluate the results of percutaneous and surgical treatment of post transplantation urinary fistulas. PATIENTS AND METHODS: 11 urinary fistulas were observed after 160 renal transplantations, corresponding to an incidence of this complication of 6.8%. Urinary fistulas were treated percutaneously in three cases and surgically in eight cases. THE FOLLOWING COMPLICATIONS WERE OBSERVED DURING SURGICAL TREATMENT: extensive necrosis of the urethra in four cases, rupture of the sutures in two cases, a punctate pelvic fistula in one case. RESULTS: The incidence of fistula was 5.8% for Leadbetter reimplantation and 8.1% for Lich Gregoir extravesical reimplantation. We obtained one success in three patients treated percutaneously and one death and seven successes out of eight patients treated surgically. We performed ureteropelvic anastomosis with the native ureter in the case of extensive necrosis of the ureter (4 cases), a new reimplantation in three cases and suture of the pelvic fistula in one case. CONCLUSION: More than one half of post-transplantation urinary fistulas observed in our department are secondary to ischaemic necrosis of the ureter. We emphasize the value of preservation of the ureteric blood supply during organ harvesting. Post transplantation urinary fistulas must be treated surgically, as soon as possible, to avoid infectious complications. PMID- 9410318 TI - [Ureteroscopy for ureteral calculi. 379 cases]. AB - OBJECTIVE: We report the experience of a centre which treated symptomatic ureteric stones exclusively by ureteroscopy. METHODS: From 1987 to 1993, 379 patients underwent ureteroscopy for ureteric stones, corresponding to 231 men and 148 women, between the ages of 8 and 80 years. The stones were situated in the pelvic ureter in 78.9% of cases, iliac ureter in 15.8%, and lumbar ureter in 5.3% of cases. An 11 F or Gautier ureteroscope was used. The only intracorporeal lithotripsy method consisted of an ultrasound transducer. Finally, our department does not possess an extracorporeal lithotripsy apparatus allowing the treatment of ureteric stones. RESULTS: The stone was eliminated immediately in 288 patients (76%), the stone was pushed back into the renal cavities in 40 patients (1.5%) and partial failures (residual fragments) or complete failures were observed in 51 patients (13.5%). 13 ureteric perforations (3.43%), treated by means of a double J ureteric stent, and one surgically drained urinoma were observed. 13 false passages of the ureteric meatus had an uneventful course by drainage with a double J stent. CONCLUSION: Ureteroscopy remains an effective method for the treatment of ureteric stones. It is associated with a definite morbidity, probably higher in a University Centre in which future urologists are trained. PMID- 9410317 TI - [Value of helical CT scan in the preoperative assessment of the ureteropelvic junction syndrome]. AB - INTRODUCTION: The search for anatomical variants of the renal blood supply is an important element in the choice of surgical technique for the treatment of ureteropelvic junction syndrome. MATERIAL AND METHODS: Prospective study of 17 patients (mean age: 33 years), assessed by spiral CT with acquisition of overlapping thin sections during the arterial phase. The renal blood supply anatomical findings were then compared with operative findings in all patients. RESULTS: We observed a solitary renal artery in 8 cases, a lower pole pedicle crossing the junction in 9 cases and vascular abnormalities in 4 cases. All of these radiological findings were confirmed intraoperatively. Spiral CT, with rapid acquisition during the arterial phase and overlapping thin sections, allows accurate analysis of the renal blood supply. Axial sections are sufficient to provide the necessary anatomical information. Three dimensional reconstructions allow a better spatial visualization of the various structures. CONCLUSION: This examination allows visualization of a possible lower pole artery associated with UPJ syndrome, with identical accuracy to that of renal arteriography, but constitutes a less invasive and less expensive investigation. PMID- 9410319 TI - [Anatomic study of the pubic-urethral ligaments in women: role of urethral suspension]. AB - OBJECTIVE: In 1963, the studies by Zacharin emphasized the importance of the suspensory apparatus of the female urethra, consisting of two non-extensible fibrous bands composed of 3 distinct ligaments. The objectives of this study were to evaluate the role of these ligaments in support of the urethra and to define the value of their preservation in cystectomy with bladder replacement in women. MATERIAL AND METHODS: Five fresh female cadavres with a mean age 78 years (range: 75-82) were used for dissection of the suspensory apparatus of the urethra via a suprapubic then sagittal approach by section of the pelvis 2 cm from the midline. Histological studies of the urethra were then performed. RESULTS: The suspensory apparatus of the urethra was found to be a paired, symmetrical system composed of 2 paired and symmetrical fibrous bands. Dissection of these bands revealed three anterior, intermediate and posterior pubourethral ligaments. The posterior pubourethral ligament was inserted on the anterior and lateral surfaces of the urethra and vagina (underneath the bladder neck) on anatomical studies, but also on histological sections. Section of the posterior pubourethral ligament allowed great mobility of half of the urethra. Section of these ligaments totally released the urethra from the pubis. CONCLUSION: Our study confirmed the importance of this ligamentous system in suspension of the urethra and demonstrated the value of preservation of these structures in cystectomy with bladder replacement in women. PMID- 9410320 TI - [Infectious risks of outpatient cystoscopy in men with sterile urine]. AB - OBJECTIVE: To prospectively assess the inherent risk of infection associated with outpatient cystoscopy performed in men with sterile urine without antibiotic prophylaxis. MATERIAL AND METHODS: 298 cystoscopies were performed in men corresponding to these criteria, with the exclusion of patients at risk of bacterial endocarditis. The equipment consisted of three cystoscopes prepared according to the recommendations of the Societe Francaise d'Hygiene Hospitaliere. The disease justifying the examination was specified for each patient. The sterility of the urine was verified during the week preceding the examination and 48 hours later by urine culture. RESULTS: Out of 281 evaluable patients, an infection was observed in 22 cases (7.8%), and was symptomatic in only one case. Escherichia coli was the organism most frequently isolated (50%) and no multiresistant bacteria were detected. A particularly high infection rate was observed in enterocystoplasty patients (21.7%). CONCLUSION: The infectious risk of cystoscopy in the presence of sterile urine, performed according to recommendations, appears to be higher than previously reported. This risk appears to be significantly higher in the case of enterocystoplasty than for other diseases. PMID- 9410321 TI - [Endoscopic treatment of residual vesico-ureteral reflux after reimplantation in children: twelve cases]. AB - OBJECTIVE: To evaluate the safety and efficacy of endoscopic treatment of vesicoureteric reflux in children persisting after surgical reimplantation. MATERIAL AND METHODS: 12 children between the ages of 17 and 103 months were treated by endoscopic injection after failure of Cohen ureterovesical reimplantation. An associated uropathy was present in 4 children. Reflux was unilateral in every case. The operation was performed as a day-only procedure. All children were reviewed at 3 months. RESULTS: No early complications were observed. The follow-up cystography was normal in 10 children. The same degree of reflux persisted in 2 children, requiring a second ureterovesical reimplantation. A meatal stricture occurred 2 years after injection. The mean follow-up was 24.7 months. The success rate of the technique was 75% in this series. CONCLUSION: This technique is reliable, simple and can be performed in the day hospital. However, these good results must not mask the risk of long-term recurrence, requiring prolonged surveillance. PMID- 9410322 TI - [Management of stage I testicular nonseminomatous germ cell tumors with an embryonic carcinomatous component. 18 cases]. AB - OBJECTIVE: To evaluate the prognosis and therapeutic modalities of stage I nonseminomatous germ cell tumours of the testis (NSGT) with an embryonic carcinomatous component (EC). MATERIAL AND METHODS: 18 patients with stage I nonseminomatous germ cell tumour of the testis with an embryonic carcinomatous component were treated between 1987 and 1995. EC represented more than 50% of the testicular tumour mass in 15 cases. This tumour contingent constituted the only potential prognostic factor in 4 cases, but vascular or lymphatic emboli (n = 3), tumour stage > pT1 (n = 5) or absence of endodermal sinus component (n = 9) were observed in 14 cases. The first 3 patients underwent retroperitoneal lymph node dissection and the following 15 patients were submitted to surveillance (n = 4) or chemotherapy (n = 11) according to the PVB [Cisplatin, Vinblastine, Bleomycin] (n = 7) or BOE [bleomycin, Etoposide, Cisplatin] (n = 4) protocols. RESULTS: With a follow-up of 10 to 110 months (mean: 46), the survival rate is 100% and the recurrence rate is 22%. None of the patients with a local stage exceeding pT1 relapsed after chemotherapy. 2 patients in whom the EC contingent represented less than 50% of the tumour mass and who were simply watched, did not relapse. 4 relapses, detected 3 to 14 months after orchidectomy (mean: 8.5), during surveillance (n = 2) or after chemotherapy (n = 2), required surgical resection or complementary chemotherapy. They occurred in patients in whom EC represented more than 50% of the testicular lesion. The tumour of initially conservatively managed patients did not contain an endodermal sinus component (n = 2) or presented vascular emboli (n = 1). The subjects treated by chemotherapy were characterized by the presence of emboli (n = 1) or the absence of endodermal sinus component (n = 1). The course after recurrence was favourable in 3 cases and the last patient is currently receiving chemotherapy. CONCLUSION: EC is an independent risk factor whose presence justifies proposal of complementary treatment by retroperitoneal lymph node dissection or chemotherapy, possibly limited to 2 courses of BOE. Surveillance can only be considered in the case of a minority of EC in the tumour, in the absence of any associated risk factors. PMID- 9410323 TI - [Ultrasonography of the urethra: prognostic value of the ultrasonographic structure of corpus spongiosum peristenotic fibrosis: prospective study of 33 patients]. AB - OBJECTIVES: The contribution of ultrasonography to pretreatment morphological assessment of strictures of the anterior urethra and assessment of the risk of recurrence after internal urethrotomy. MATERIAL AND METHODS: 33 patients (16-89 years) operated by internal urethrotomy for stricture of the anterior urethra and followed for at least 6 months. Preoperative urethral ultrasonography, recording the number, length and degree of strictures and echostructure of the peristenotic fibrosis of the corpus spongiosium. RESULTS: Ultrasound visualization of all urethral strictures, with no false-positives and no false-negatives. 11 patients relapsed after a mean interval of 5.7 months (1-16 months), 22 patients did not present recurrence: mean interval: 15.5 months (6-36 months). Corpus spongiosum fibrosis associated with urethral stricture is isoechoic to the corpus spongiosum (19 cases) or hyperechoic to the corpus spongiosum (14 cases). No statistical correlation was observed between the echostructure of the fibrous tissue and the risk of recurrence after internal urethrotomy. CONCLUSION: Ultrasonography allows excellent analysis of the morphological characteristics of a stricture of the male anterior urethra. In our experience, and in contrast with the limited data of the literature, no correlation was observed between the echostructure of the peristenotic fibrosis and the risk of recurrence after internal urethrotomy. PMID- 9410324 TI - [Acucise endoureterotomy for distal ureteral stenosis in a renal transplant patient]. AB - The treatment of ureteric strictures in renal transplantation used to be surgical, but has recently benefited from progress in endourology. The authors report the case of a renal transplant recipient who developed late stricture of the ureterovesical reimplantation of the transplant. After percutaneous nephrostomy, which restored good renal function, retrograde endoureterotomy was performed using an Acucise ureterotome balloon, followed by ureteric modelling on a 7F double J stent for 2 months. With a follow-up of 18 months, renal function was normal and ultrasonography showed residual hypotonia of the transplant cavities and no vesicorenal reflux was detected by retrograde voiding cystourethrography. Acucise retrograde endoureterotomy can constitute a simple endourological treatment for late ureteric strictures in renal transplantation. PMID- 9410325 TI - [Renal angiomyolipoma with venous extension associated with complete duplication]. AB - A case of venous extension de renal angiomyolipoma is reported. The diagnosis was established preoperatively by ultrasonography and renal CT. This tumour was treated conservatively by partial nephrectomy after selective embolization because of the presence of complete duplication. PMID- 9410326 TI - [Anterior anal sphincter approach in the management of urethral-rectal fistulas: a case report]. AB - Urethrorectal fistula is a rare disease which is easy to diagnose, but which essentially raises a problem of the choice of the most appropriate surgical technique. A review of the literature on the subject shows that, over the last decade, various teams have developed the anterior and posterior trans-anp sphincteric approaches. We report a case of an acquired urethrorectal fistula associated with anal incontinence treated via an anterior trans-ano-rectal perineal incision. The exposure provided by this incision facilitates simultaneous cure of the fistula and repair of the anal sphincter. PMID- 9410327 TI - [Laparoscopic resection of a bladder diverticulum]. AB - The authors report a case of laparoscopic resection of a bladder diverticulum performed at the same time as endoscopic treatment of its presumed cause. Dissection of the diverticulum, particularly its neck, and dissection of diverticula of the ureter and vas deferens are facilitated by magnification of the video image. This approach is only indicated in particular anatomical sites (posterior). The results of this technique must be compared with those of conventional treatments. PMID- 9410328 TI - [Prostate cancer screening (II): is prostate cancer a public health problem? Update of incidence and mortality figures in France from 1982 to 1990]. AB - OBJECTIVE: To describe the epidemiological situation of prostatic cancer in France on the basis on a large population sample. MATERIAL AND METHODS: This study uses incidence data derived from French cancer registers, and mortality data obtained from death certificates. Crude rates and rates standardized for the world population are calculated. The variation of these rates is analysed by a log-linear model (adjusted for age, department and period). RESULTS: The incidence of prostatic cancer in France in 1990 was 200 to 300/100,000 between the ages of 60 and 70 years and more than 600/100,000 after the age of 70 years. 73% of cases were diagnosed after the age of 70 years. The incidence increased annually by 8.76% between 1982 and 1990. An estimated 22,600 cases were diagnosed in France in 1990. The increased incidence of localized or local stages is due to the use of diagnostic tests (PSA and ultrasound-guided biopsies), as this increase accelerated after 1987. The crude mortality rate was 33.4/100,000 (384/100,000 between the ages of 75 and 85 years). It increased by 2.56% per annum from 1982 to 1990, but essentially for men over the age of 75 years. CONCLUSION: These findings tend to make prostatic cancer a public health priority, but this affirmation must be moderated by the fact that this disease has a low impact on loss of life expectancy. PMID- 9410329 TI - [Prostate cancer screening (III): risk factors, natural history, course without treatment. Characteristics of detected cancers]. AB - The Oncology Committee of the Association Francaise d'Urologie has up-dated the knowledge concerning prostatic cancer screening since the 1989 Consensus Conference. The results are published in the form of a series of articles referring to the criteria used as prerequisites for cancer screening programmes. This article reports the data of the literature concerning risk factors, natural history, course without treatment and histological characteristics of the cancer detected. 1) Certain populations have an increased risk due to genetic factors. A family history (first degree relative) is associated with a 2- to 3-fold higher risk of prostatic cancer. This familial aggregation can be used to define a high risk group constituting a primary target for screening. 2) The natural history of the disease, especially the progression from the asymptomatic stage to the clinical stage and the natural history of the disease at the clinical stage are now sufficiently well known. The concept of latent cancer has not been confirmed as the disease inevitably progresses. Cancers of insignificant volume, less than 0.5 cc (discovered at autopsy) are classically distinguished from cancers of significant volume, greater than 0.5 cc, but asymptomatic (risk of progression with mortality within 15 years), and local and/or metastatic symptomatic cancers. The histological prevalence of prostatic cancer is 43% in a group of men with a mean age of 64 years and increases with age. 92% of histological cancers have a volume less than 0.5 cc. It takes an estimated 12 years (3 doubling times) for a 0.5 cc cancer to reach a volume of 4 cc, the volume beyond which there is a risk of distant metastases. In the absence of curative treatment, a cancer diagnosed at the localized stage before the age of 65 years is associated with a specific survival of less than 30%. The median survival of metastatic prostatic cancer is 2 to 3 years. 3) The disease can be detected at an early stage. Cancers diagnosed by an isolated elevation of PSA in a screening setting have a significant volume in more than 3 out of 4 cases, can be entirely removed by prostatectomy in more than 3 out of 4 cases and have a less advanced pathological stage than cancers diagnosed on the basis of classical criteria. PMID- 9410330 TI - [Arguments against long term use of complete androgenic blockers]. PMID- 9410331 TI - [Inguinal lymphadenectomy in cancer of the penis: surgical techniques and indications]. AB - Lymph node invasion is one of the major prognostic factors of cancer of the penis. However, as it is difficult to evaluate clinically and by means of complementary investigations, inguinal or even ilioinguinal lymph node dissection is still indicated. As this surgery carries a certain morbidity (necrosis of skin edges, infection, lymphorrhoea and subsequent lymphoedema), the indications are presented according to the presence or absence of palpable inguinal lymph nodes and the stage of the primary tumour. Various surgical techniques are proposed: Superficial and deep inguinal lymph node dissection in the case of mobile and palpable inguinal nodes, simplified and superficial inguinal lymph node dissection in the absence of palpable inguinal nodes and in the case of invasive primary tumour. PMID- 9410332 TI - [Nosocomial urinary infections]. AB - The concept of nosocomial urinary tract infection now corresponds to a precise definition. It is generally related to bladder catheterization, constitutes the most frequent form of nosocomial infection (30 to 50% of infections), and represents the third most frequent portal of entry of bacteraemia. The organism most frequently isolated is Escherichia coli; but the flora is changing and the ecological distribution is continually modified. Despite their usually benign nature, these nosocomial infections can nevertheless influence hospital mortality; they increase the hospital stay by an average of 2.5 days and their treatment represents a large share of the antibiotic budget. Prevention of these infections is therefore essential, with particular emphasis on simple and universally accessible measures: very precise indications for vesical catheterization, use of closed circuit drainage, maximal asepsis when handling catheters, after washing the hands. PMID- 9410333 TI - [Rehabilitation of female urinary incontinence. Techniques and indications]. AB - Female uro-gynaecological retraining was initially confined to prophylactic management during the post-partum period. It has gradually been extended to other diseases, either alone or as a complement to medical or surgical treatment. The results of clinical examination and complementary investigations, especially urodynamic studies, now allow the application of increasingly precise retraining techniques. This progress essentially concerns the three types of technique most frequently used: Manual retraining concerns the bulbospongiosus muscles, as well as the levator ani muscles. Biofeedback is increasingly used to qualitatively improve muscle contraction. Electrostimulation can promote the action of various groups of tonic or phasic muscle fibres. Apart from these endocavitary techniques, retraining is also evolving towards a global management of the abdominopelvic sphere., where it is indicated in the management of the post partum period and various gynaecological and urinary diseases. However, concomitant diseases such as perineo-abdominodiaphragmatic imbalance, an anorectal problem, a sexual problem or finally painful symptoms, must also be taken into account in some cases. Retraining must be very precisely prescribed, confirming application of all of these techniques and adapting them to the national health refund classification. The number of sessions varies according to the disease, but rarely exceeds 30 sessions in the initial prescription. On the other hand, like any muscle system, the pelvic floor must be maintained by means of several sessions per year. The success of this retraining treatment essentially depends on several factors: the quality of the therapist, his training, his spirit of integration in a multidisciplinary team, but also the patient's motivation. PMID- 9410334 TI - [The lymphatic system and its functioning in sheep]. AB - Cannulation of the afferent and efferent lymph ducts of a lymph node in large domestic species have greatly facilitated the study of the immune system functions in mammals, both in physiological conditions and during immune responses to various pathogens. Summarizing the numerous studies undertaken in the ovine species, this review first presents a description of the structure and composition of the lymph nodes, vessels as well as the lymph fluid they transport. The functioning and the role of each of these immune compartments are also discussed. The characteristics and mechanisms of lymphocyte recirculation between the blood and the lymphatic system are treated in an other chapter. The last part is dedicated to analyzing the modifications observed in the ovine lymphatic system after antigenic stimulation. PMID- 9410336 TI - Policy ... privacy of electronic medical records. PMID- 9410337 TI - Earnings. Just say grow. PMID- 9410335 TI - [Bovine hypodermyiasis: a quasi sequential procedure for observation of live stock for surveillance of recrudescence in eradicated zones]. AB - A quasi-sequential sampling plan for observing cattle was undertaken in the framework of Hypoderma bovis epidemiological surveillance. This plan enabled us to test whether or not the prevalence of Hypoderma bovis is within a threshold p0 chosen for an eradication scheme or for qualifying a zone. The sampling cost was reduced but the estimation precision remained acceptable. The sensitivity of monthly controls with respect to the reference period (April, May, June, July) was studied from data observed over 6 consecutive years in Cotes d'Armor, France. The best ratio sensitivity/sampling cost was obtained with a unique counting plan in June where 48% (36 to 60%) of the herds found to be infested during the reference period were detected. The test procedure concerning the prevalence level was then adapted to take into account the sensitivity of the observation method. This is tantamount to reducing the threshold p0. PMID- 9410339 TI - Management ... managed care will begin looking more and more like the old indemnity system. PMID- 9410340 TI - Medicare ... navigating the Medicare system. PMID- 9410338 TI - Finance ... managed care organizations. PMID- 9410341 TI - Litigation ... hospitals alleged to be bilking Medicare. PMID- 9410342 TI - Nursing ... you ask a nurse, right? PMID- 9410343 TI - Physicians ... data on doctors is fair game. PMID- 9410345 TI - Technology ... build an information system. PMID- 9410344 TI - Religion ... line is drawn between physical and spiritual healers. PMID- 9410346 TI - Litigation. Call this one a draw. PMID- 9410347 TI - Reimbursement ... New York hospitals. PMID- 9410348 TI - Litigation ... Medicare billing probe. PMID- 9410349 TI - Access ... rationing of care. PMID- 9410350 TI - Integration ... number of U.S. physicians. PMID- 9410351 TI - Compensation ... group practices. PMID- 9410353 TI - [Age and in vitro fertilization. French National Register on In Vitro Fertilization]. AB - The mean age of IVF patients women and men is always increasing, this is probably due to the late desire for pregnancy nowadays. Pregnancy rate decreases with the age of wife and in a minor way with the age of man. The most important factor is the quality of woman answer to ovarian stimulation. Among women of 42 or over, the one with a high estradiol level on hCG day (> 4000 pg/ml) are the only older women with a correct pregnancy rate. PMID- 9410352 TI - [FIVNAT 1996 report. French National Register on In Vitro Fertilization]. AB - In 1996, the French National Register on in vitro fertilization, FIVNAT, has collected information on 32,490 IVF oocyte recoveries, from which 65.0% were conventional IVF cycles and 34.6% were ICSI. No modification was observed concerning the women's age. On the opposite men's age increased slightly. A small increased was observed in the pregnancy rate per recovery (20.5%) and per transfer (26.0%). The fertilization rate (49.2%) was at the same level than in 1995. The same was true for the mean numbers of oocytes but the mean number of embryos increased at 4.14 +/- 4.0. The mean number of transferred embryos significantly decreased to 2.53 +/- 0.95 per transfer. For conventional IVF, the percentage of infertilities of tubal origin was 51%, whereas almost 40% of the recoveries involved a male factor. The percentage of inseminations realised with a donor's semen, which had decreased in 1995, remained low in 1996 (2.6%). The stimulative regimen involved a GnRH analogue mostly with a long blockage period (78.6%) which was associated to the highest pregnancy rate. Only 12.8% of the transfers involved more than 3 embryos, and were associated to a relatively poor pregnancy rate, confirming that they were preferently realised in couples with poor chances of pregnancy. PMID- 9410354 TI - [Results after an in vitro fertilization pregnancy. French National Register on In Vitro Fertilization]. AB - For this analysis, all the oocyte recoveries realised between 1989, January 1st and 1995, December 31th and collected by the French national register on IVF (FIVNAT) were included. Three groups were constituted, depending on the existence of a previous birth (group 1, n = 8,615), or of a previous failed pregnancy (spontaneous abortion or ectopic pregnancy, group 2, n = 8,539), both obtained by IVF, or no previous IVF pregnancy (group 3, n = 139,843). In this last group, women and men were of younger age (respectively 33.3 +/- 4.7 vs 35.1 +/- 3.9 for group 1 and 34.3 +/- 4.5 for group 2, p < 0.001, and 35.2 +/- 5.9 vs 37.0 +/- 5.1 for group 1 and 36.1 +/- 5.6 for group 2, p < 0.001). Overall results were lower in the group 3. The average number of embryos was 3.7 +/- 3.7 compared to 4.4 +/- 3.8, p < 0.001 for the two other groups. The fertilization rates was at 47.3 +/- 33.2% in group 3, compared to 55.5 +/- 30.5 in group 1 and 55.6 +/- 30.4 in group 2 (p < 0.001). Finally, the per recovery pregnancy rates were respectively 17.9%, 28.0%, 25.0% (p < 0.001). In group 3, infertility origin was less often due to a tubal disease (53.2% vs 65.6% in group 1 and 66.6% in group 2, p < 0.001). On the opposite, an isolated male origin was more frequent (respectively 19.9%, 11.0% and 12.2%) (p < 0.001). In group 3, compared with those with secondary infertility, women with primary infertility were younger (32.6 +/- 4.6 vs 34.7 +/ 4.6, p < 0.001), had more oocytes (8.9 +/- 5.7 vs 8.1 +/- 5.4, p < 0.001), but less embryos (3.6 +/- 3.7 vs 3.9 +/- 3.7, p < 0.001), a decreased fertilization rate (44.8 +/- 33.4 vs 51.6 +/- 32.6, p < 0.001) and a decreased pregnancy rate (17.3% vs 18.9%, p < 0.01). Infertility was more often of male origin (23.6% vs 11.7%, p < 0.001). PMID- 9410355 TI - [1996 report of in vitro fertilization with a third donor]. AB - 1996 results of French federation of CECOS and IFREARES-Toulouse are recorded. In 1996 13% less AID demands were registered. 4,132 patients underwent procreations with sperm donor during 11,793 cycles (AID and IVF D), resulting in 1,366 pregnancies. 386 volunteers men come forwards as sperm donors (40.3% less than 1995). The analysis of the 1,266 deliveries resulting of 1995 activity show always the same percentage of multiple births but an increase in intra-uterine AID malformations. The activity of semen preservation is increasing mainly for testicular cancer. PMID- 9410356 TI - [Paternal age and fertility]. AB - Variations of the male partner fertility according to his age are difficult to study, because of bias represented by some decrease of both sexual activity and fecundity of the female partner beyond 35 years old. The few studies which are available show that sperm concentration as percentage of normal forms, are stable with age. The percentage of motility is the only parameter which decreases systematically with age according to all studies. Nevertheless, sperm fertilizing capacity does not seem to be decreased. FIVNAT data allow to analyse almost 30,000 IVF cycles for tubal sterility; they confirm the absence of an important alteration of semen characteristics with age. Despite a significant decrease- even moderate--of the mean fertilization rates, the pregnancy rates remain roughly constant for a given range of maternal age. PMID- 9410357 TI - [Seasonal variation in sperm characteristics]. AB - In all species, seasonal patterns in fertility are observed. In humans, seasonal variations of pregnancies are observed in spontaneous reproduction or in IVF. The most important factor seems to be sperm values and specially the morphology. Our study, on 188,075 cases (FIVNAT 1987-1995) shows significant variations of semen characteristics: we observe, in August, an improvement of sperm morphology. PMID- 9410358 TI - [Evolution of male fertility in the last twenty years]. AB - After the meta-analysis by Carlsen et al. (1992) showing a decline in human sperm count over the last 50 years, several studies have been made on the sperm characteristics of more or less homogeneous groups of men who were collecting semen in the same center for 10-20 years. A significant decline in sperm count was reported in some studies but not in others. The debate on declining sperm counts is not closed and these studies raised the following important questions: could the differences in results be the reflect of variation in techniques or blas in methodologies? If these phenomena are real, why a deterioration of semen quality in some places and not in others with so important geographical differences of sperm production? What are the possible consequences on human fertility and what are the causes? Unfortunately, there is presently no answer. The danish study provoked extensive discussion on the many possible sources of bias while there was less controversy concerning the studies carried out in a single center despite the extent of limitations of several of them. Recent publications indicated concomitant and increasing alterations of the development and/or function of the male genital tract, various observations in the wildlife and several experimental studies suggesting the possible deleterious role of numerous chemical compounds present in our environment. Therefore, prospective epidemiological studies and fundamental researchs are urgently needed. PMID- 9410359 TI - [Assisted reproduction and viral hepatitis: the virologist's viewpoint]. AB - Medical assistance to procreation in a couple with one of the parents having viral hepatitis raises the issue of the transmission of infection to the baby and of possible contaminations of gametes from virus-free parents in the laboratory. Today, the main problems are linked to hepatitis B and C viruses, which induce chronic disease transmissible to the baby and can be transmitted through laboratory contaminations. PMID- 9410360 TI - [Assisted reproduction and hepatitis C virus infection]. AB - Medically assisted procreation poses a difficult problem when one or both partners of couple are infected with HCV. Epidemiologic and fundamental works show a low risk of HCV sexual transmission and no pregnancy complications or fetal abnormalities have been reported. However, the outcome of HCV infected children is unknown. These contrasting findings suggest that medically assisted procreation using sperm of spouse needs to be cautious. Before medically assisted procreation, testing of couples for HCV antibodies must be done and interferon therapy is required for patients with histological chronic active hepatitis and for HCV positive mothers or infected couples. In the absence of specific legislation or consensual recommendations, detailed informations must be given to the couples in order to obtain an informed consent. PMID- 9410361 TI - [In vitro oocyte growth and maturation: a mouse example]. AB - During the last 20 years, numerous studies have been performed to improve culture conditions in order to allow in vitro growth and meiotic maturation of oocytes isolated from primordial to antral follicles. The main objective of these studies is to define optimal culture conditions which will allow to increase the recovery rate of oocytes that can be successfully fertilized. This paper reviews the more recent advances obtained when studying mouse oocyte development in vitro. PMID- 9410362 TI - [In vitro folliculogenesis: a woman is not a mouse!]. AB - The informations obtained using in vitro folliculogenesis in rodents have shown that ovulation of preantral follicles or granulosa-cumulus complexes can be obtained after a few days of in vitro culture and that live young can be obtained following fertilization and development of these oocytes. In addition, initiation of follicular growth from primordial follicles could be achieved in organ culture in vitro. Hence, when the two techniques were combined, all folliculogenesis (from the primordial stage to ovulation) could be achieved in vitro. The situation is more complex in primates/human owing to the lower number of follicles initiating growth per day and to the longer duration of folliculogenesis. A few reports suggest that preantral follicles can survive and grow in culture for a few days and one study demonstrates that in 2 months, follicles can grow from 150 microns to 1.6 mm in diameter. The short term interest of such studies is to improve our understanding of the mechanisms involved in basal folliculogenesis. In the longer term, such techniques may prove useful to maintain some reproductive potential to women submitted to total body irradiation, to improve the management of reproduction of women suffering from early menopause or to generate oocyte banks to donate oocytes. PMID- 9410363 TI - [In vitro murine spermatogenesis: cytologic study of the meiosis stage]. AB - Spermatogenesis is a complex process of cellular multiplication and differentiation, the regulation of which is only partially understood. This may be due; 1) to the redundancy of the mechanisms of local regulation; 2) to the ubiquitous character of those growth factors and cytokines present in the testis; 3) to the lack of long term culture systems allowing male germ cell development. We describe herein two culture systems associating Sertoli cells and germ cell in which part of the meiotic step can occur in vitro. In the first system, pachytene spermatocytes prepared by centrifugal elutriation are cultured on a layer of Sertoli cells. In the second system small fragments of seminiferous tubules are seeded in bicameral chambers. The cytological and immunocytochemical studies presented show that pachytene spermatocytes of stages IV-VI differentiate into spermatids of steps 1 to 5. PMID- 9410364 TI - [Paracrine control of spermatogenetic stem cells: example of the leukemia inhibitory factor]. AB - Correct regulation of spermatogonial mitosis and, more specifically, the control of the balance between differentiation and proliferation to allow renewal of the stem cell stock, are essential for the maintenance of spermatozoa production throughout life. The mechanisms underlying this control are still unknown. However, recent studies suggest that some locally produced cytokines may be involved in the regulation of spermatogonial activity. In this context, Leukemia Inhibitory Factor (LIF) exhibits interesting properties regarding stem cells and, particularly, primordial germ cells. The present study aimed at investigating LIF production and LIF binding abilities by/of the different testicular cells types (somatic and germ cells). Our study demonstrates that LIF is produced within the testis, mainly by peritubular cells which are in the vicinity of spermatogonia, the latter cells expressing high levels of LIF receptors. These results strongly suggest an involvement of LIF in the control of spermatogonial activity. PMID- 9410365 TI - [Role of tumor necrosis factor in the male gonad]. AB - Within the male gonad, TNF alpha is secreted by macrophages in the interstitial tissu and by germ cells in seminiferous tubules. TNF alpha receptors (type I) have been detected in the somatic cells of the testis (i.e. Leydig and Sertoli cells). TNF alpha has two type of action within the testis: the cytokine has an anti-hormonal role and play a role in the interactions between Sertoli and germ cells. Anti-hormonal actions of TNF alpha are (i) inhibition of LH-induced testosterone production in Leydig cells and (ii) inhibition of FSH-induced inhibin production in Sertoli cells. Moreover, TNF alpha; produced by germ cells, enhanced expression of growth factors in Sertoli cells (IGFBP3, receptor type I of bFGF) and enhanced production of energetic metabolite important for germ cells such as lactate (via enhancement of lactate dehydrogenase enzyme, isoform 5). Moreover, Fas/Fas ligand system, proteins related to TNF alpha, seem to play a key role in the protection of germ cells against the immune system within the testis. In conclusion, TNF alpha and related proteins, could play a great role in testicular functions such as spermatogenesis (interactions between Sertoli and germ cells) and in the interactions between immune system and spermatogenesis (role of TNF alpha originating form blood/testicular macrophages, role of Fas/Fas ligand system). PMID- 9410366 TI - [Acute salpingitis: current antibiotic protocols]. AB - Nowadays, most patients with uncomplicated acute salpingitis undergo ambulatory treatment. The choice of medications must take features of PID into account: they are mult-microbial infections and a prolonged follow-up is necessary in order to decrease the risk of sequellae. To fulfil these goals, combination of antibiotics are prescribed that are active against C. trachomatis, enterobacteria and anaerobes. PMID- 9410367 TI - [Laparoscopy and salpingitis. For whom? When? How?]. AB - The apparent decrease of the incidence of pelvic inflammatory disease (PID) in industrial countries comes from the increase of less symptomatic forms. Diagnosis requires more often non clinical diagnostic technics. Laparoscopy remains the gold standard for the diagnosis and grading of PID but its indications should be on an accurate strategy. PMID- 9410368 TI - [Post-infectious tubal infertility: indications for assisted reproduction]. AB - PID is the major cause of tuboperitoneal infertility. Management of tuboperitoneal infertility associate surgery and IVF the choice between these two technique is depending of the stade of the tubes evaluated by HSG, salpingoscopy and laparoscopy. PMID- 9410369 TI - [Identification and treatment of endometritis]. AB - Endometritis represent a debatable entity because genital infectious disease is a contre-indication of endoscopy. So, the diagnosis is rarely realized by hysteroscopy. Two kinds of endometritis are discerned: acute endometritis characterized by an oedema of bleeding endometrium, covered by an abnormal mucus; chronic endometritis with areas of red endometrium, flushed, with a white central point, localized or scattered out the cavity. This is the "strawberry aspect". Whereas, the correlation between hysteroscopic aspect and histologic or bacteriologic samples exists in only 35% of cases. According to R. Frydman and J. Hamou, chronic endometritis is occurred among 22% of patients in IVF program, in 14% of unexplained infertility and 23.6% of women with an history of first trimester miscarriages. Chlamydiae an Ureaplasma seem to be the most frequent germs. In this way, in case of unexplained infertility, the hysteroscopic diagnosis of chronic endometritis can lead to an antibiotic treatment test. PMID- 9410370 TI - [The infected cervix]. AB - Cervico-vaginal infections while easily identified, can in certain cases, in fact quite frequently, be under-diagnosed because of the absence of visible symptoms. Colposcopic access to the cervix or vagina can aid diagnosis of cervico-vaginal infections but may not be adequate to pinpoint the agent of the infection except in certain cases such as Herpes and HPV. On the other hand, cervical-vaginal infections can impede a colposcopic interpretation such as identifying the squamo columnar junction or the location of a directed biopsy. Cervical infections can equally cause unexpected complications (notably Haemorrhagie or pelvic infections) during or after surgical Colposcopic procedures. PMID- 9410371 TI - [Current requirements of sterilization and disinfection of endoscopic materials]. AB - Bacterial and viral infectious risks related to gynecologic endoscopy and laparoscopy are a significant problem. Contamination between patients or by environmental bacteria can occur when sterilisation or disinfection procedures are inadequate. Endoscope steam sterilisation is the safest method and have to be employed whenever possible. For heat labile materials, disinfection procedure will comply with the recommendations stated in a recent ministerial decision, including a cleaning step with a non aldehydic detergent-disinfectant and a disinfection step with 2% glutaraldehyde. Rooms and equipment dedicated to disinfection have to be well adapted to this use and specific staff training must be achieved. Endoscope automatic washer disinfections are an alternative method to manual disinfection but they have to meet precise criteria to insure safety and efficacy. PMID- 9410372 TI - [Luteal phase support after vaginal progesterone: comparative study with micronized oral progesterone]. AB - Two progesterone presentations, a vaginal application of 90 mg progesterone per day (Crinone) or 300 mg progesterone orally administered (Utrogestan) were compared for luteal phase support of patients undergoing an in vitro fertilisation (IVF) procedure. 283 patients were randomly allocated to either treatment. The treatment started within 24 hours after the embryo transfer procedure and continued until day 30 in cases of implantation. Efficacy was assessed using the pregnancy and delivery rates. Safety was assessed through specific symptoms and usual safety monitoring. The pregnancy rate per transfer was not significantly different in the Crinone and Utrogestan groups at day 12 (Crinone: 35.3%, Utrogestan: 29.9%, p = 0.55), at day 30 (Crinone: 28.8%, Utrogestan: 25%, p = 0.61), at day 90 (Crinone: 25.9%, Utrogestan: 22.9%, p = 0.69). No difference in spontaneous abortion rates were seen thereafter. The delivery rate was not significantly different (proportion delivery per patients included, Crinone: 23.0%, Utrogestan: 22.2%, p = 1), as well as the ratio new born per transferred embryo (Crinone: 11.7%; Utrogestan: 11.1%, p = 0.91). Safety parameters were similar in both groups except for drowsiness, which was more significantly frequent in the oral P group than in the Crinone group at all time points. No serious adverse events were recorded in this study. The fact that Crinone matches the efficacy of the larger doses of progesterone used orally reflects an advantage of the transvaginal route of administration which avoids metabolic inactivation of progesterone during its first liver pass. PMID- 9410373 TI - [Practice of intra-cytoplasmic sperm injection in France: report of the BLEFCO study]. PMID- 9410374 TI - [Are there indications for intra-cytoplasmic sperm injection in female infertility?]. AB - ICSI for oocyte pathology may represent the solution in case of zona pellucida anomalies, deficiency of the oolemma fusion ability or absence of cortical reaction. These indications remain scarce and difficult to be displayed. A link could be shown by some authors between some oocyte morphologic anomalies and IVF fertilization failures which are overcome by ICSI. Other dysmorphic oocytes are associated with an impairment in cytoplasm function which results in a poor embryonic viability on which ICSI has no effect. When few oocytes are recovered (< or = 5) and the semen characteristics are not optimal, ICSI may rectified the decrease in fertilization rates obtained after classical insemination. Lastly, ICSI will become increasingly used in case of oocyte in-vitro maturation or freezing instead of classical IVF which is less efficient. PMID- 9410375 TI - [Intra-cytoplasmic sperm injection with testicular sperm: what strategy?]. AB - The pregnancy rate after ICSI with surgically retrieved spermatozoa varies according to the etiology and to the level of sperm retrieval in the genital tract. The use of testicular sperm in all cases is discussed. PMID- 9410376 TI - [The value of donor sperm in intra-cytoplasmic sperm injections? The French Centers for Semen Preservation policy]. AB - Some IVF laboratories ask for donor's semen during an ICS attempt if retrieval of husband's sperm is unsuccessful. The French Centers for Semen preservation (CECOS) consider that this request is not acceptable for legal, psychological and ethical considerations. Legal consent may not be ambiguous. The psychological lost of biological fatherhood should be totally assumed before undertaking conception with donor's semen. ICSI should be started only if it is most likely to succeed. For all these reasons, CECOS refuse to modify the strategy of procreatione during the course of an attempt. PMID- 9410377 TI - [Y chromosome and spermatogenesis]. AB - World wide about 2% of men have a primary deficiency in sperm production. Familial cases of male infertility and chromosomal anomalies associated with this phenotype indicate that there is, at least in some cases, a genetic cause. In particular, deletions of the non-recombining portion of the Y chromosome are associated with a failure of sperm production. Deletion screening, using Y specific markers has defined three regions (AZFa, b, c) that are associated with azoospermia. Each region contains one or more candidate genes that may be responsible for the phenotype. However, mutations have not yet been identified in any of these genes. This review discusses the structural organisation of the human Y chromosome and genes that are candidates for male infertility. PMID- 9410378 TI - [Clinical elements for the diagnosis of secretory azoospermia]. AB - Secretory azoospermia is defined by the existence of azoospermia, normal epididymis and vesicular markers and often high but sometimes low FSH levels. Study of the past history often reveals the cause of azoospermia, which is often toxic or a side effect of medication. Clinical investigation includes assessment of the genital organs in particular the volume of the testicles. The clinical investigation must always be accompanied by a general examination. When anamnesis and clinical examination are completed, secretory azoospermia will be classed in one of the following categories: high FSH azoospermia, law FSH azoospermia, normal FSH azoospermia. PMID- 9410379 TI - [Management of testicular biopsies in secretory azoospermias]. AB - Management of testicular biopsies in non-obstructive azoospermia is unclear: there is non-factors that are predictive of eventual sperm presence or absence and there is no consensus neither for open testicular biopsy versus percutaneous testis biopsy nor for fresh testicular sperm extract ion and synchronous ICSI versus frozen samples for a later sample. PMID- 9410380 TI - [Psychosocial and sexual outcome in women with Turner syndrome]. AB - Results of an inquiry in adults patients (18-53 years of age) coming from three centers (1 in Rouen & 2 in Paris), 213 questionnaires were sent. 105 answers were received. Scholar achievement: only secondary cycle in 6%, Secondary cycle + professional course in 22% and tertiary cycle in 44%. No scholar ship in 2%, 26% were still ongoing studies. Professions: 18% are unemployed (24%), Secretary jobs: 10%, Health professions (altogether): 18%, Teachers: 8%, Clerks: 7%, Executive jobs: 8% and Miscellaneous jobs: 5%. 3% have an handicapped status. The small height was a career obstacle in 29%. Affective life. Age of first sexual intercourse was 19-22 years. 17 are or were married and 15% are living in couple. But 58% have not any sexual life whatsoever. These women are divided on the ways to cure the sterility. Few among the oldest have attempted adoption or medically assisted procreation, with each time low rate of success. 26% have psychological disturbances which were serious in 6% mainly due to depression. PMID- 9410381 TI - [Autoimmune precocious menopause: therapeutic consequences]. AB - In case of spontaneous, idiopathic, karyotypically normal premature ovarian failure (POF), autoimmune disease's characteristics have to be verified: association with other autoimune diseases, initial psychological stress, circulating antibodies to thyroid, adrenal or the ovaries, particular HLA haplotypes. Ovarian follicular activity can be measured by ultrasonography. Estrogentherapy is able to enhance ovarian activity. The potential benefit of corticosteroid therapy needs further evaluation. PMID- 9410382 TI - [Results of oocyte donation in women with different indications, ages and therapeutic strategies]. AB - PURPOSE: Compare the effectiveness of oocyte donation for various indications, ages and treatments. METHODS: Retrospective analysis of 319 transfers for 184 couples (243 with fresh embryos and 76 with frozen embryos). Transfers are divided into: -7 groups according to indication: premature ovarian failure (POF) with normal caryotype n = 122, surgical POF n = 26, postchemotherapy and postradiation therapy POF n = 23, genetic disease carrier n = 11, IVF failure n = 69, primary amenorrhea n = 58, POF with abnormal caryotype n = 10. -2 groups according to the recipient's age (Group I < or = 40 years, Group II > 40 years); 3 groups according to the dose and the mode of administration of the treatment. Oocytes were donated by volunteer donors of less than 38 years having at last one child. RESULT: According to the recipient's age 31.3% clinical pregnancies (CP) and 24.2% ongoing pregnancies (OP) for group I, versus 19.4% CP and 11.9% OP for group II. The results according to the indication were similar except for postchemotherapy and postradiation therapy POF for which the rate of CP as well as the rate of OP were particularly low. For the treatment groups there is a significant reduction of CP and OP as the treatment dose increases. CONCLUSION: The recipient's age plays an important role for the development of pregnancy and this shows that apparently uterine ageing also plays an undeniable role for the decline of female fertility. Increasing doses of estradiol reduces the rate of CP and OP. Postchemotherapy and postradition therapy POF has much lower results than other indications. PMID- 9410383 TI - [Can we define the indications for oocyte donation?]. AB - The outline of oocyte donation's indications is defined in France by the law of the 29 july 1994. A great number of situations do not pose any problem with regard to this principles: gonadal dysgenesis, premature ovarian failure, ovariectomy, castration following chimiotherapy or radiotherapy. However, others indications drive us progressively outside legal limits. What is the normal age of physiological menopause? Which threshold to justify oocyte donation for low responders? Is oocyte donation an alternative for other treatments failures? Ovocyte donation for genetic reason presents a risk of eugenic drift. Answering these questions is uneasy, though it is urgent for our patients to offer them a better uniformity. PMID- 9410384 TI - [Legal aspects of oocyte donation]. AB - Oocytes donation is managed by two bioethic french laws. We will have a look at the enforcement of these in comparaison with their non specificity. PMID- 9410385 TI - [Oocyte donation: psychological and sociological aspects, the clinician's viewpoint]. AB - Among the treatment for medical assisted procreation, oocyte donation presents the more complex implications on psycho-sociological aspects. French law of the 29th July 1994, tries to prevent risk for the principal actors of oocyte donation, but has created new problems: 1) the shortage of donors lengthens the waiting list and reduces the succes's chance, 2) the absence of uniformitybetween different countries opens the door to an international market of the oocyte. Medical progress give new possibilities for situations that presented no problem in the past, for instance, pregnancies after menopause. Natural markers are questionned, it is too soon to conclude and to mesure all the consequences of our treatment on the well-being of the child, the recipient's couple, and the donor. PMID- 9410386 TI - [Brief psychoanalytic point of view on assisted reproduction]. AB - Some psychological mechanisms, regarding the new techniques of ART are reported in this work. A short overview of Freudian theory of psychosexual development of the child is given. The narcissistic wound induced by the diagnosis of sterility is quite important but quite different for men and women. It seems that oocyte donation may "cure" fantasmatically some female sterilities, given that this technique allows some women to get pregnant and to deliver a child, which was until then out of their biological "fate". Whether the anonymity of gamete donation is adapted to the psychic structure of recipients, donors, and medical teams is a question that should be considered. Very few works have been reported about the fantasies of the medical teams on this topic. The importance of infantile sexual theries is stressed again. PMID- 9410387 TI - [Problems posed by oocyte donation organization]. AB - Ten years after the first pregnancy in France and 3 years after the setting of the law of Bioethic oocyte donation procedure is coming up against many difficulties (search of donors, cost, efficacy). We will try to consider all of them to prepare the way for an official recognition of this specific procedure of AMP. PMID- 9410388 TI - [Psychological consequences of triplet births: evaluation at four years]. PMID- 9410389 TI - [Laparoscopy and ovarian cancer: dangers and precautions]. PMID- 9410390 TI - [Thyroid cancer and pregnancy (literature review and a case report)]. PMID- 9410391 TI - [Is there still a role for laparotomy in tubal infertility surgery?]. PMID- 9410392 TI - [Sex offenses: hormonal treatment]. PMID- 9410393 TI - [Cytomegalovirus infections and pregnancy]. PMID- 9410394 TI - [Abnormal endometrial reactivity to colony stimulating factor 1 and leukemia inhibitory factor dependent female infertility]. AB - Maternal LIF is essential for embryo implantation in mice and it may also be the case in humans. We recently reported that endometrial LIF secretion from infertile women presenting repeated failures of embryonic implantation or unexplained sterility was significantly lower than the secretion of explants from fertile women. We now report on the modulation of the endometrial LIF secretion according to the obstetrical status. CSF-1 has little effect or increases LIF secretion from fertile women whereas it inhibits secretion from infertile women with repeated failures of embryonic implantation. PMID- 9410395 TI - [Revisiting GnRH analog protocols II. Strategy for poor responders]. AB - In the first part of this review, we analyzed protocols for ovarian hyperstimulation with GnRH analogs and stressed both their diversity and uncertainty for mechanisms which lead to improve the outcome of IVF cycles. Apart from their beneficial effects on prevention of endogenous LH surges and premature luteinization, other adverse effects have been reported: need of higher amount of exogenous gonadotrophins, direct negative action on ovarian function and conceptus quality. They must be considered before defining a strategy for poor responders to IVF protocols using GnRH analogs. The recent trend for reducing GnRH doses must be evaluated. PMID- 9410396 TI - [HIV virus detection in ejaculates collected at different times in seropositive patients]. AB - Ejaculates from 22 seropositive males were collected at various times 1 to 11 ejaculates for each in one year period of time. After fractionation, no virus is detected in mobile and living spermatozoa. Moreover, HIV is not always found in all ejaculates from the same patient. PMID- 9410397 TI - [Constipation]. AB - Constipation may be due to drug intake, metabolic and endocrine abnormalities, neurogenic diseases, and gastro-intestinal lesions: in these cases, constipation is only the consequence of another disease. Constipation may be due to dietary or behavioural errors in a lot of patients: in these cases constipation is the consequence of the way of life of the patients. When these two first types of constipation have been ruled out, the constipation is a true constipation, described as chronic idiopathic constipation for which no organic cause may be found. This chronic constipation may be due to a delayed colonic transit secondary either to a colonic inertia in few cases or, at the opposite, to an increased colonic motility in a lot of people often associated with abdominal pain. Chronic constipation may be also due to an outlet obstruction, secondary to a megarectum, a megarecto-sigmoid, a rectocele, or an abnormal anorectal motility (mainly anismus, often associated with sexual abuse in the past history). No single mechanism can be held responsible for chronic idiopathic constipation, but, whatever the mechanism and the type, constipation is also frequently a way to get help from the doctor. PMID- 9410398 TI - [Failure of the anti-abortion movement in France]. PMID- 9410399 TI - [The use of carbon carriers for the immunoenzyme diagnosis of viral infections]. AB - Carbon solid phase for enzyme-linked immunosorbent assay was proposed. This support can be used instead of polysterene plates because the following steps (carbon support incubate with specific and detective reagents) are implemented in arbitrary minimal capacities. The method was tested for detection of the virus antigens (hepatitis B, herpes simplex) and anti-viral (anti-adenovirus, anti influenza virus) antibodies. PMID- 9410400 TI - [The use of the fluorescent probe method for the diagnosis of acute coronavirus intestinal infections in people]. AB - The scientific investigations conducted first allow using a method of fluorescent probes for the diagnosis of human coronaviral intestinal infection. The method is characterised by sensitivity, specificity, reproductivity and proximity. It is comparable, according to these parameters, with the methods of the third generation and may be recommended for putting into practical work of virological laboratories. PMID- 9410401 TI - [The structural-functional organization of the diphtheria toxin]. AB - The present data on the structure and functional features of diphtheria toxin, the main pathogenic factor of diphtheria infection, have been described. Information on the primary and secondary structures and X-ray analysis of this protein is presented and discussed. The structures of catalytic, transmembrane and receptor-binding domains of diphtheria toxin have been considered in detail and the functional roles of some amino-acid residues of these domains have been analyzed. PMID- 9410402 TI - [Surgical treatment of idiopathic macular hole: what is new one year later]. AB - We report the results of 23 idiopathic macular hole surgeries on 23 patients consecutively operated on. There are 20 stage III and 3 stage II holes, according to the classification of Gass. Anatomical success, defined as a macular hole with a 360 degrees flat edge, or as a macular hole which is no more visible, is obtained in 13 cases; functional success, defined as a visual acuity improvement of 2 lines or more at the last visit, is obtained in 11 cases. Postoperative rhegmatogenous retinal detachment is noted in 2 cases. With such results, our conclusions are less optimistic as in 1994. If macular hole surgery is very efficient in some patients, larger studies are still needed in order to better define the most successful surgical indications and technique. PMID- 9410403 TI - [Efficacy of pneumatic retinopexy: 7 year retrospective study]. AB - We describe the results of pneumatic retinopexy in the treatment of primary rhegmatogenous non traumatic retinal detachment in 63 patients with a follow-up of 6 months or more. We compare these results with those of other pneumatic retinopexy series and series of pneumatic retinopexy-eligible cases treated with alternative methods. We conclude that pneumatic retinopexy is a method of first choice in the treatment of a large number of retinal detachments, taking into account the final anatomic and functional success rate, the simplicity and cost effectiveness of the technique, the patient comfort and very early functional rehabilitation. PMID- 9410404 TI - [Transcanalicular dacryocystorhinostomy by pulse Holmium-YAG laser]. AB - We report the results of 26 Holmium-Yag transcanalicular DCR made in 23 outpatients, under topical anaesthesia. Success rate was 47% (follow-up: 1 to 7 months). Failures occurred within the first month. PMID- 9410405 TI - [Recurrent corneal erosions: new manifestation of juvenile alport syndrome]. AB - A history of recurrent corneal erosion (RCE) in 3 brothers with juvenile X-linked Alport's syndrome (AS), whereas their 2 brothers without AS were unaffected, suggested a possible association between AS and RCE. A survey, among 39 patients with AS, revealed that 6 of them had suffered from RCE, supporting the association of the two conditions. RCE has to be considered as an additional ocular manifestation of AS, with a prevalence of 15 to 21%. PMID- 9410406 TI - [Internuclear bilateral pseudo-ophthalmoplegia and dermatomyositis]. AB - A 60 year-old woman complaining of diplopia presents an ocular motility disturbance mimicking internuclear ophthalmoplegia. Idiopathic dermatomyositis is diagnosed by the help of clinical, biological, electrophysiological and histological data. The outcome is favorable under corticotherapy. Ocular muscle involvement is rare in dermatomyositis. An overlap syndrome with another auto immune disorder like myasthenia should be excluded in this kind of manifestation. PMID- 9410407 TI - [A case of painful bilateral ophthalmoplegia]. AB - The authors present a case of Tolosa-Hunt syndrome with the particularity of a simultaneous palsy of both oculomotor nerves. The evolution is followed during 16 years. The clinical description, the criteria of the syndrome and the differential diagnosis are discussed. The diagnosis is based on the clinical evolution, the transitory palsy of the oculomotor nerves and the involvement of the optic nerve. The response on steroid treatment was spectacular. Bilateral and fluctuant oculomotor palsy is possible in the TolosaHunt syndrome. General investigations are mandatory to exclude other pathologies. PMID- 9410408 TI - [Strabismic amblyopia: management and visual acuity]. AB - This study presents the clinical history of 9 patients presenting an inversed amblyopia during the course of their treatment. A simple therapy was sufficient to restore the initial sensorial situation for all cases in a less than two months period. Two of these patients were presenting a so strong dominance of fixation that they were still fixing with their eye, even after becoming amblyopic. The use of the Gracis's biprismatic test is considered as an objective witness of the quality of the dominance and is illustrated through these examples. This test can make the difference between an inversed amblyopia and a real inversion of the initial amblyopic process. It permits to choose the type and the duration of the treatment using to prevent the recurrence of amblyopia. PMID- 9410409 TI - [Amblyopia from anisometropia without strabismus]. AB - Fifty non-strabismic children with primary anisometropia were reviewed retrospectively. At entry, patients ranged in age from 1 to 10 years with an average of 4.5 years. The follow-up ranged from 0.5 to 9 years with an average of 3.5 years. Criteria for inclusion were a difference in refractive error between the two eyes of at least 1.00 D of spherical value and/or 0.75 D of cylindrical value. In all cases, anisometropia was totally corrected by prescribing glasses. Anisometropic amblyopia was considered to be present when isoacuity at far was not reached despite the glasses, part-time occlusion therapy of the good eye was prescribed. Amblyopia was present in 86% of the patients and was found with all types of anisometropia. It was more severe in anisohyperopia and/or anisoastigmatism. After adequate treatment, amblyopia was clinically cured or less severe in 78% of the patients. PMID- 9410410 TI - [Amblyopic strabismus: strategy for late treatment]. AB - We show the result of the late treatment of 11 esotropic patients. The initial amblyopia has conditioned the analysis of our results. We defend the use of prolonged monocular occlusion, relayed by an optical penalization at far consisting on an hyperopic overcorrection of +2.50 D. This study confirms that the efficacy of the treatment depends principally on its compliance. We want to underline the danger of interrupting the occlusion too soon and without supervision. PMID- 9410411 TI - [Peripheral ulcerative keratitis and a form of limited wegener's granulomatosis]. AB - A man aged 68 years presents superior limbal infiltrates at his left eye two weeks before a marginal ulcer which quickly perforates. He has no systemic complaint. Clinical, biological, radiologic and histological evaluations disclose superior airways and lungs implications, an inflammatory syndrome, high ANCA (antineutrophiles cytoplamic antibodies) titer and vasculitis. There is no sign of renal involvement. A limited form of Wegener's granulomatosis is diagnosed. The outcome is favorable with a partial penetrating keratoplasty and systemic corticosteroid therapy in association with immunosuppressive drugs. This so called limited form of Wegener's granulomatosis is sight threatening when eye is the initial presentation. The early diagnostic and treatment will be performed by the help of ANCA in cases with subclinical systemic manifestations. PMID- 9410413 TI - Monitoring Pressure Ulcer Healing: An Alternative to Reverse Staging. Proceedings of the National Pressure Ulcer Advisory Panel 5th National Conference. 1997. PMID- 9410412 TI - [Lacrimal-nasal canal stenosis after intranasal surgery]. AB - The etiology of the nasolacrimal duct obstruction usually reported is: idiopathic, infectious, traumatic, congenital, tumoral and others. The nasolacrimal duct stenosis after intranasal surgery is rarely reported in the literature. We describe 5 cases of iatrogenic nasolacrimal duct stenosis found in our series of 41 patients treated the last 2 years for nasolacrimal duct obstruction and we propose how to avoid this complication of intranasal surgery. PMID- 9410414 TI - [The effect of spasmolytics on dilatation of the uterine cervix]. AB - Spasmolytics and spasmoanalgetic mixtures are administered to facilitate dilatation of the cervix during delivery and to shorten the first stage of labour. This medication is used in 70% deliveries at the 1st Dept. of Obstetrics and Gynaecology Masaryk University in Brno. The aim of the study was to analyze the spasmolytic effect on the process of cervix dilatation. The study comprises 108 uncomplicated deliveries (52 deliveries with spasmolytics and 56 deliveries without). We analyzed the effect of spasmolytics on the cervix, process of labour and outcome of delivery. Six kinds of spasmolytics were used, administered from once to four times during labour and most of them (85%) in the latent phase of the first stage of labour. In the group with spasmolytics there was statistically insignificant by prolonged active phase of the first stage of labour (126 vs 104 minutes) and more frequent administration of Oxytocin. Others parameters such as length of the second stage of labour, rate of birth injuries, the 1st minute Apgar score less than 7 and the necessity of neonatal resuscitation did not differ in the two groups. CONCLUSIONS: The application of spasmolytics did not significantly affect the process of delivery. PMID- 9410416 TI - [Pregnancy and delivery after frozen embryo transfer during a cycle without a corpus luteum and an artificially prepared endometrium by administration of a GnRH analog]. PMID- 9410415 TI - [Adrenergic tocolytics--their effects on lipoperoxidation in the brain]. AB - As a sequence of our previous investigations and published results dealing with the inhibitory effect of the BETA-mimetic Tocolyticum (Partusisten) on the blood plasma levels of polyunsaturated fatty acids (n-3), we performed experiments where the possible inducing effect of BETA-mimetic drugs (epinephrine and isoprenaline) on the lipoperoxidative processes in the brain and liver was studied. (Adult rats-females.) Lipoperoxidation was tested by the method of Ohkawa and al. [21] based on changes in malondialdehyde (MDA) production. The intraperitoneal administration of isoprenaline (0.15 mg/kg body weight) increases significantly the MDA production in the cerebral cortex as well as in the medulla oblongata of rats (females-Wistar). Isoprenaline induces a small but significant decrease of MDA in the liver. The intraperitoneal administration of epinephrine (0.15 mg/kg body wight) raises significantly the MDA production in the cerebral cortex as well as in the medulla oblongata. In hepatic tissue changes in MDA levels were not observed. The mentioned data indicate that the decrease of polyunsaturated fatty acids (esp. of n-3 series) in plasma of human newborns and their mothers treated by BETA-mimetic tocolytic drugs (as Partusisten), could be associated with the significant higher lipoperoxidation. The clinical use of such tocolytic drugs has to be discussed also from this point of view. PMID- 9410417 TI - [The effect of a broad spectrum antibiotic on the incidence of postoperative complications]. PMID- 9410418 TI - [Relaxation-suggestion-hypnotic methods in labor]. PMID- 9410419 TI - [Malignancies in pregnancy]. PMID- 9410420 TI - [The effect of spermatozoal ultrastructure on the outcome of fertilization in vitro]. AB - The ultrastructure of sperm cells in cumulus oophorus of the human oocyte 20 hrs after in vitro fertilization (IVF) was studied. MATERIALS AND METHODS: Mechanically removed cumular cells were processed for electron microscopy and the spermatozoa deposited in the intracellular spaces as well as those penetrating into follicular cells were evaluated. The samples were divided into three groups. Group I contained cumuli from unfertilized oocytes, group II involved those from fertilized oocytes, but not pregnancy after embryo transfer. Group III was formed by cumuli from fertilized oocytes of patients, where pregnancy had been achieved. RESULTS: In group I numerous anomalies of spermatozoal ultrastructure were found, such as head malformations, incompleted chromatin condensation, large cytoplasmatic droplets, etc. In groups II and III, about 70% normal sperm cells were found. CONCLUSIONS: These findings provide evidence of the important role of the morphological quality of spermatozoa in the failure of in vitro fertilization, also in cases where anomalies could not be detected by the usual methods of sperm examination. PMID- 9410421 TI - [New trends in the management of endometrial carcinoma]. PMID- 9410422 TI - [Classification, terminology and surgical technique of laparoscopic hysterectomy]. PMID- 9410423 TI - [The effect of hormonal contraception on liver function]. PMID- 9410424 TI - [Study of urinary incontinence in women using perineal and introital sonography]. PMID- 9410425 TI - [Profile of antiphospholipid antibodies in various diagnoses associated with reproduction]. AB - The authors investigated antiphospholipid antibodies (APA) against phosphatidic acid, ph-ethanolamine, ph-DL-glycerol, ph-inositol, ph-L-serine and cardiolipine in isotypes IgG and IgM in various diagnoses associated with reproduction. They found that the most varied pattern of high levels of these antibodies is in particular in patients with the diagnosis of intractable infertility (in 26.5% women IgG against cardiolipine, in 24.5% patients IgG against ph-inositol and ph DL-glycerol, in 17.6% women IgG against ph-ethanolamine, in 20.6% women IgG against phosphatidic acid antigens). Intractable infertility, repeated failures of IVF and patients with a basic autoaggressive disease were followed up and their antiphospholipid antibodies were assessed. In one patient follow up of APA revealed SLE before the patient developed clinical symptoms of the disease. PMID- 9410426 TI - [Dosage of acetylsalicylic acid in the prevention and therapy of intrauterine growth retardation]. AB - The authors recommend, based on their own experience, an optimal dose of acetylsalicylic acid in treatment of IUGR. The therapeutic effect was not proved conclusively and views are controversial. Based on a retrospective group from the years 1993, 1994 and 1995 the authors assume that treatment of IUGR by acetylsalicylic acid is indicated and they recommend a dose of 100 mg per day (e.g. one tablet of Anopyrin). PMID- 9410427 TI - Janice Weinberg's career clinic. Your references: important allies in your job campaign. PMID- 9410428 TI - Stephanie Burgess, President of the SCNA on 2-1-96 before the SC Health & Human Service Finance Commission. PMID- 9410429 TI - The Board of Nursing adopts CDC recommendations. PMID- 9410430 TI - [Old and new "vaccines" in respiratory diseases]. PMID- 9410431 TI - [Compliance with antismoking laws in official institutions]. AB - The prevention of nicotine addiction involves a wide range of measures, including writing laws to preserve public health by protecting nonsmokers from smoke and discouraging smokers from consumption. Also important are campaigns to educate both parties (smokers and nonsmokers) about the negative effects of tobacco. The main antismoking law in Spain is the Health and Consumer Ministry's Royal Decree 192/1988 limiting the sale and use of tobacco with the aim of protecting public health. Other regulations have since been enacted by public administrations to complement that law. Research finding published in recent years have been the basis for major legal changes leading in two directions; toward standardizing laws existing in different countries and toward increasing restrictions on the advertising and sale of tobacco. Various scientific and social groups have demanded that current laws be made stricter. Little has been done, however, to assess the degree of vigilance and compliance, and consequently the efficacy, of current legislation. The aim of this study was to determine the level of compliance with the law in governmental institutions in Salamanca. We visited 30 centers and saw that while notices prohibiting smoking were visible in 80%, the number of smokers was high: 43% among workers (none of whom was in educational or medical centers) and 37% among the public. No posters warning of the dangers of tobacco were seen in any of the centers visited. It appears necessary to further restrict the sale and use of tobacco in public places, to enforce compliance with existing regulations and to increase the amount of information on the toxic effects of tobacco in order to gain the cooperation of both smokers and nonsmokers toward achieving smoke-free environments. PMID- 9410432 TI - [Induced sputum in asthma: study of validity and repeatability]. AB - To assess the validity and reproducibility of determining total and differential cell (DC) counts and of eosinophilic cationic protein (EPC) levels in induced sputum. Clinical analysis of validity and reliability of a measurement tool. Twenty-one asthmatics [age: 31(14) years; FEV1: 78(21)% of reference value; PC20FEV1: 0.02(1.1)mg/ml] and 10 healthy subjects [age: 30(3) years, FEV1: 99(12)% of reference value, PC20FEV1: > 8(3.7)mg/ml]. A sputum sample was collected from each individual using hypertonic saline. Sputum was separated from saliva and then divided into two portions as uncontaminated as possible by squamous cells. Both portions were treated independently and simultaneously within two hours with a solution of bithiothreitol to disperse the cells, and then centrifuged. The supernatant was poured off and frozen al -70 degrees for later determination of EPC. The sediment was stained with Papanicolaou stain, toluidine blue and eosinhematoxylin. DC count was expressed as a percentage of total inflammatory cells. ECP was determined with a CAP-System and a commercial kit (Pharmacia Diagnostics SA, Uppsala, Sweden). Two subjects in each group were unable to produce valid sputum samples (20% of healthy subjects and 10% of asthmatics). The validity of the method was demonstrated by significant differences between asthmatics and healthy subjects in both eosinophil and EPC levels. The reproducibility of method was verified by comparing the results for the sputum fractions; no significant differences were found in cell counts or EPC levels. The within-group correlation coefficients for the sputum fractions ranges between 48 and 77% for all cell types, except squamous cells, which gave a coefficient of 18%. For EPC the correlation coefficient was 98%. DC and EPC determinations in sputum induced by hypertonic saline are valid and reproducible. PMID- 9410433 TI - [Comparative evaluation of 3 types of syringes routinely used for arterial gas analysis]. AB - This prospective study evaluates the practical utility of a special plastic syringe for collecting arterial blood samples for gasometry, comparing it to a glass syringe and a conventional plastic one, in 120 patients who came to our hospital for arterial blood analysis for a variety of reasons. The patients were randomly assigned to one of three groups according to type of syringe assigned. Technicians with experience in the technique prepared the syringes and collected the arterial blood samples after providing local anesthesia. The results showed that the special plastic syringe took less time (p < 0.05) to prepare than did the glass or conventional plastic ones. The differences were quite small, however, in absolute terms, with the special syringe requiring 17 seconds less than the glass syringe and 6 seconds less than the plastic one; the practical importance is therefore slight. No other advantages of the special syringe were observed. No patient required more than one puncture to obtain a valid arterial blood sample, the extraction times (time between arterial puncture until end of process) were similar in the three groups of patients, and the presence of post puncture hematoma was rare in all groups. There were no differences in level of pain reported (on an analog scale) and the subjective quality of the radial pulse wave was good and similar in all three groups. In conclusion, these results show that use of the special syringe offers no important practical advantages for experienced technicians that would justify the higher price. In fact, as the conventional plastic syringes are cheaper, disposable and similarly effective in expert hands, our results suggest that their routine use for collecting arterial blood gas samples can help improve the cost-benefit ratio for a common procedure in pulmonary function units. PMID- 9410434 TI - [Production of tumor necrosis factor alpha and interleukin-6 by alveolar macrophages from patients with rheumatoid arthritis and interstitial pulmonary disease]. AB - The aim of this study was to measure the production of tumor necrosis factor alpha (TNF-alpha) and interleukin-6 (IL-6) by alveolar macrophages in patients with rheumatoid arthritis and interstitial lung disease (ILD). Rheumatoid arthritis patients diagnosed by ACR criteria (n = 8) with associated ILD documented by pulmonary function tests and high resolution computerized tomography scanning, and 12 healthy volunteers (6 smokers and 6 nonsmokers). We determined the spontaneous and induced production of bacterial lipopolysaccharides (LSP), TNF-alpha and IL-6 by alveolar macrophages obtained by bronchoalveolar lavage. The macrophages were isolated by Ficoll-Hypaque gradient centrifugation and plastic adherence and cultured in serum-containing medium (low endotoxin) in the presence and absence of LPS (100 ng/ml). TNF-alpha and IL-6 levels contents were determined in supernatants by ELISA. In the patient group both spontaneous and induced production of TNF-alpha were significantly higher than in controls (p < 0.01). Macrophages stimulated with LPS in patients with rheumatoid arthritis and ILD also released greater amounts of IL-6 than did those of the healthy controls. IL-6 spontaneous and induced production was significantly lower in smokers than in nonsmokers. TNF-alpha and IL-6 production in patients with rheumatoid arthritis and ILD, studied in bronchoalveolar lavage specimens reveals that alveolar macrophages are hyperreactive in these patients, who are possibly sensitized as a consequence of the inflammatory lung process inherent to the disease. Further study is needed to define the pathogenic role of these mediators. PMID- 9410435 TI - [Videothoracoscopy versus thoracotomy in the diagnosis of diffuse interstitial disease]. AB - Histopathology plays an important role in the diagnosis of diffuse interstitial lung disease. New endoscopic techniques allow performance of sampling procedures that have the same diagnostic accuracy and fewer complications that open lung biopsy (OLB). We evaluated our experience with video thoracoscopic lung biopsy (VTLB) in comparison with OLB in terms of diagnostic accuracy, duration of pleural drainage and hospital stay, number and size of samples and complications. Thirty patients who underwent VTLB from March 1987 to January 1995, and 28 patients who underwent OLB from May 1987 to January 1995 were studied retrospectively. Diagnosis was achieved in the VLTB group in 96.66% of cases and in the OLB group in 92.85%. The number of specimens obtained was 1.5 (SD 0.5) in the VTLB group (p = 0.47). Overall specimen size expressed in cm3 was 13.3 (SD 15.4) in the VTLB group and 18.5 (SD 21) in the OLB group (p = 0.284). Length of pleural drainage in hours was 52 (SD 39.9) in the VTLB group and 89.1 (SD 47) in the OLB group (p = 0.002). Length of hospital stay in days was 3.5 (SD 2) in the VTLB group and 8.7 (3.5) in the OLB group (p = 0.001). Complications were fewer and less severe after VLTB. VLTB is as useful as OLD for obtaining lung biopsies in diffuse interstitial lung disease. VLTB also causes fewer complications and is less expensive because drainage times and hospital stays are shorter. PMID- 9410436 TI - [Diagnosis and treatment of nosocomial pneumonia]. PMID- 9410437 TI - [Magnetic resonance in lung carcinoma]. PMID- 9410438 TI - [Right upper lobar mass of the lung caused by subclavian artery aneurysm]. AB - Subclavian artery aneurysm is a rare condition. The main causes are degenerative disease and, less often, trauma. We report the case of a sawmill worker with a large mass in the upper right lobe found in a routine X-ray. Imaging studies revealed the aneurysm to be 12.2 x 13.1 cm, partially thrombosed and located in the right subclavian artery. Our experience suggests that this cause of lung mass should be considered early in the diagnostic process, before undertaking invasive diagnostic (puncture-biopsy) or therapeutic procedures that might place the patient at risk. PMID- 9410440 TI - [An unusual case of posterior mediastinal mass: extraosseus Ewing sarcoma]. PMID- 9410439 TI - [Schwannoma of the intrathoracic vagus nerve]. AB - Schwannomas, or neurilemomas, are tumors that originate in the sheaths covering peripheral nerve fibers. They are usually encapsulated, slow growing, and asymptomatic. Such tumors may appear in any nerve, although most are found in the extremities. Intrathoracic vagus schwannomas are very rare and only 72 cases have been reported. We present a new case in a 39-year-old man in whom chest film showed a well-defined, homogeneous mass measuring 3 cm in diameter located in the left para-aortic region. The presence of a smooth tumor on the vagus nerve and recurrent loop was confirmed by left lateral thoracotomy. After removal, the tumor was shown to be a vagal nerve schwannoma. PMID- 9410441 TI - [Primary, bilateral, and synchronous pulmonary double tumor. Report of a case]. PMID- 9410442 TI - [Recurrent hemoptysis as presentation form of saccular aneurysm of the thoracic aorta. Report of a case]. PMID- 9410443 TI - [The need for presenting in the conclusions of clinical trials the limitations and possible biases of their design and evaluation]. PMID- 9410444 TI - [Sustained-release ++theophyllins of 12 and 24 hours. Comparative study]. PMID- 9410445 TI - [Perception of the Central Hospital by the urban population of Yaounde: quantitative and qualitative methods]. AB - Two studies, using different methods, were used to assess the opinions of the local population on Yaounde Central Hospital (YCH), the changes made as part of a restructuring program and use of health care facilities. The study population comprised YCH patients and the urban population of Yaounde as a whole. One study used personal interviews, according to standard epidemiological survey methods (1,267 inhabitants). The other used focus group discussions involving 8 to 12 people each (10 groups). There was no significant difference in the results recorded by these two methods. Focus group discussions are cheaper and easier to implement and are thus valuable for use alone or prior to more extensive studies using questionnaires. Classic epidemiological survey methods produce results which are reproducible and suitable for statistical analysis, and will make it possible to follow the progress of the new working policies of the YCH. PMID- 9410446 TI - [Gynecology-obstetrics at the Yalgado-Ouedraogo (Ouagadougou) National Hospital Center. Cancer of the cervix uteri: epidemio-clinical and anatomopathologic aspects]. AB - We report the findings of a 3-year retrospective study aimed at describing the epidemiological, clinical, anatomical and pathological profile of cervical cancers in an African country. We studied 46 cases of invasive cervical cancer. This type of cancer accounts for 31.7% of female genital cancers. The mean waiting time for a consultation was 7 +/- 2.4 months. The average age of the patients was 48 +/- 3.7 years. They were mostly women who had had several pregnancies without paid work. The main reasons for seeking medical help were metrorrhagia (95.6%), pelvic pain (58.7%) and purulent discharge (45.6%). In 89.1% of the cases, the cancer was inoperable. Pathology results were available in 37 cases. We found 36 cases (97.3%) of epidermoid carcinoma and a single case (2.7%) of adenocarcinoma. There were associated condylomatous lesions in 21.6% of cases. These results demonstrate the importance of making cervical smear tests a routine part of medical examination and of making the public aware of the importance of screening for cervical cancer. PMID- 9410447 TI - [Gynecology-obstetrics at the Yalgado-Ouedraogo National Hospital Center. Eclampsia: epidemiologic, clinical and prognostic aspects]. AB - We present the results of a retrospective study carried out between 1992 and 1995 aimed at describing the epidemiological, clinical and developmental profile of eclampsia in an African maternity unit towards the end of the 20th Century. The incidence of eclampsia was 108 cases in 12,175 births (0.89%), mostly in young patients during their first pregnancy. 40.7% of the patients were less than 20 years old and 59.3% were expecting their first child. Eclampsia occurred between the 28th and 37th weeks of amenorrhea in 37% of cases. Thirty four patients (31.5%) had had at least three episodes of eclampsia prior to admission. Diastolic arterial blood pressure was higher than 120 mmHg in 25.9% of cases. Eclampsia occurred before labor in 30.6% of cases, during labor in 38% of cases and after giving birth in 31.5% of cases. Postpartum episodes occurred an average of 67 +/- 18.7 hours after the birth. There were complication with infection in 7 cases, renal insufficiency in 14 cases and one case of retro placental hematoma. Seventeen patients died, giving a death rate of 15.7%. During the same period, 3.4% of maternal deaths were due to eclampsia. The perinatal mortality rate was 23.1%. A quantitative and qualitative improvement in prenatal consultations should make it possible to reduce the incidence of eclampsia. Measuring arterial blood pressure daily for at least 14 days after the birth appears to be necessary for diagnosis and treatment of all cases of hypertension. PMID- 9410448 TI - [Anthropometric characteristics of HIV infected malnourished children in Ivory Coast]. AB - Stunting and weight-loss are common complications of HIV infection in children. The aim of this study was to assess whether stunting, assessed using a height-for age index, is a discriminating factor for HIV in malnourished children. This is a retrospective study of 66 children, all older than 15 months, with marasmus-type malnutrition. They were studied at a nursery in the Cote d'Ivoire from 1994 to 1995. Forty-five percent of the children were HIV-positive. The anthropometric indices (weight-for-age, weight-for-height, height-for-age and body mass index) were lower in seropositive than in seronegative children, but the difference was not significant. The rate of stunting was similar in the two groups, with 66% of seropositive and 58% of seronegative children stunted. Thus, stunting was not a discriminating factor for HIV infection. Studies aimed at increasing our understanding of nutritional disorders associated with HIV are necessary to improve the nutritional management of these children, especially in Africa where malnutrition is endemic. PMID- 9410449 TI - [Prevention and control of malaria in pregnant women in an urban setting (Yaounde, Cameroun)]. AB - The aim of this study was to evaluate the methods of preventing malaria (chemoprophylaxis, vector control) and of fever management (presumptive treatment of malaria) used for pregnant women in Yaounde, Cameroon and to identify the most important factors for assessing these practices. The 221 women studied were selected by cluster sampling. All had made extensive use of health services during pregnancy and 77% were using chemoprophylaxis. The number of febrile episodes in pregnant women who claimed to have used chemoprophylaxis was not significantly different to that in the women who did not use it. However, the mean birth weight of the babies of women who had used chemoprophylaxis was significantly higher. The women did not systematically use vector control measures; 21% used insect repellents (electric plaques, coil burners) and 20% used aerosol insecticides. Only 10% of the women slept under simple, untreated mosquito nets and none used mosquito nets impregnated with insecticide. Fifty per cent of the women had at least one episode of fever during pregnancy and 77% were treated for presumed malaria. However, the treatment was not standardized and was unsuitable in a third of cases. Possible changes in the chemoprophylaxis strategy are discussed. PMID- 9410451 TI - [Tetanus in Libreville: hospital analysis of 30 cases]. AB - We investigated 34 cases of tetanus in patients younger than 75 years old in Libreville. We found that the incidence of tetanus was 8 cases for every million people. The disease mostly affected people younger than 50 (85%). Only two neonatal tetanus cases were reported. The death rate was 74%, despite the availability of intensive care and respiratory assistance. We stress the benefits of tetanus immunization, which must be continued, and suggest the use of intrathecal serotherapy for the cure of tetanus in Libreville. PMID- 9410450 TI - [Ultrasound diagnosis of fetal malformations in utero: apropos of 30 cases]. AB - This study assess the incidence of fetal malformations and the contribution of ultrasound scans to their diagnosis in utero. We reviewed the files of 1,960 pregnant women of whom 30 (1.53%) had malformed fetuses. Ultrasound scanning detected 29 of these cases and failed to detect only one case (an arterial canal malformation). Neurological malformations, particularly hydrocephalus and anencephaly, were most common (12 cases), followed by urinary tract abnormalities (hydronephrosis and renal polycystosis). Other malformations, such as those of the digestive tract, thorax and face, occurred less frequently. Ultrasound scans can be used to decide whether the pregnancy should be continued and what postnatal treatment should be given. PMID- 9410452 TI - [Human intestinal microsporidiosis in Bamako (Mali): the presence of Enterocytozoon bieneusi in HIV seropositive patients]. AB - A study was conducted between 1993 and 1996 in Bamako to determine the rate of occurrence of microsporidia in 88 patients. Most (80%) had chronic diarrhea associated with weight loss and 87.5% were HIV-positive. Intestinal microsporidia were detected in 32% of the patients infected with HIV-1, HIV-2, or coinfected with both strains. Microsporidiosis was also diagnosed in three of the eleven HIV negative individuals (27%). Microsporidiosis was confirmed by electron microscopy in 6 HIV-positive patients and 1 HIV-negative individual. Enterocytozoon bieneusi was detected in each case. These results suggest that microsporidia are common pathogens in HIV-positive patients in Bamako. Cases of microsporidiosis have been reported for the first time in HIV-2-infected patients. The proportion of women microsporidiosis patients is higher in Mali than in industrialized countries. The presence of microsporidia in HIV-negative patients suggests that these parasites may be an underestimated cause of enteritis in developing countries. PMID- 9410453 TI - [Impact of changes in the environment on vector-transmitted diseases]. AB - We have defined the relationship between infectious diseases and environmental conditions and considered the development of this relationship to its current situation, where human intervention is occurring more often and is becoming more aggressive. The increase in the transport of freight and passengers by air has allowed parasite vectors to spread quickly and easily over large distances. Every country can now be reached from any other country within a couple of days. Usually, foreign species are unable to establish themselves and to persist in the new environment; but the recent arrival of Aedes albopictus in Albania, Italy and the Americas is a cause for concern. Demographic pressure has increased the need for land and the exploitation of new areas leads to large changes in the vegetation. The classic example of this man-made damage is the destruction of tropical forest in Western Africa, but the destruction of herbaceous vegetation, such as papyrus, in East Africa, could also have serious epidemiological consequences. Streams and rivers have been managed for power production and irrigation. The use of dams, both large and small, and the culture of rice in paddy-fields produces large expanses of water which are suitable breeding grounds for mosquitoes and snails, the vectors of human diseases such as malaria and schistosomiasis in sub-Saharan Africa. They are, however, of lesser importance in Asia and the Americas. Urbanization imposes a set of very similar structures on a specific rural environment. The effect of these two factors on each other determines the pathologies associated with each town. The suburban area is a specific environment where both urban and rural diseases occur and are made worse by poor hygiene conditions (waste, sewage, etc.). However, not all man-made changes to the environment cause a deterioration in public health. Urban and agricultural development projects must consider these issues and should use medical and environmental studies to avoid causing epidemic-prone conditions or spreading endemic diseases. Currently, most studies are limited to listing the specific diseases in the target area and very few attempt to assess the possible consequences of changing the environment. Forecasting the consequences of changes in environmental management is of great importance, but it requires the development of multi-disciplinary teams in the field who must be involved in the planning and implementation of the projects. PMID- 9410455 TI - Resource allocation: beyond evidence-based medicine and cost-effectiveness analysis. PMID- 9410454 TI - [The dual role of tumor necrosis factor (TNF) during a malarial attack]. AB - The roles of the various factors implicated in the pathogenesis of severe malaria are not well understood. Tumor necrosis factor (TNF), a cytokine produced mainly by macrophages, seems to play a crucial role in both the host's defence against the parasite and the development of severe complications. This review investigates the dual role of TNF in acute malaria, summarizing current knowledge of the beneficial and detrimental effects of this molecule. Recent work has suggested a possible explanation for this dualism, involving a complex interaction between TNF and its soluble receptors. PMID- 9410456 TI - Can we provide evidence-based care for the elderly? PMID- 9410457 TI - [Cytological parameters of buccal epithelium for the diagnosis of functional state of man]. AB - The functional status of humans may be safely, rapidly, and objectively assessed by evaluating the functional, morphological, and cytogenetic characteristics of buccal epitheliocytes. Three categories of the functional status were distinguished in students aged 19-20. The first category was denoted as the norm or health and comprised 30% of examined students. The second category is borderline state, comprising 53% examinees. The third category is risk group or predisease, to which 17% of students were referred. This method is recommended for population screenings. PMID- 9410458 TI - [Comparative analysis of various methods of determination of fibrinogen in the plasma]. AB - Fibrinogen was measured in the plasma of normal subjects and patients with various hemostasis disorders by gravimetric methods with thromboplastin coagulation and coagulation with Echis multisquamatus snake venom. Coagulation was assessed using optic Coag-Mate HM (Netherlands) and roll Amelung KC4A (Germany) coagulometers during coagulation with thrombin after Klauss and with the above venom. The results were compared with those of fibrinogen measurements by radial immunodiffusion using Behring antifibrinogen serum. The data of all methods coincided if the initial levels of fibrinogen were normal, whereas in manifest hypo- or hyperfibrinogenemia Klauss' method was the most accurate and informative. PMID- 9410459 TI - [Laboratory diagnosis of bacterial vaginosis]. AB - Microbiological analysis of discharge from the genitals in passenger car conductors (5787 women and 1496 men aged 20 to 40) showed Gardnerella vaginalis to be the most frequent agent of urogenital infection. It was isolated in 26% cases, whereas Trichomonas were isolated in 2.5%, fungi in 1.5%, and gonococci in 0.27% cases. The diagnosis of gardnerellosis is reliable if the key cells are found. Monoinfection with G. vaginalis was diagnosed in 80% patients, the overwhelming majority (74%) of carriers of this bacterium had no inflammatory symptoms, and in only 26% the carrier state was associated with the presence of leukocytes and histiocytes, more often in the cervix. In men the carrier state was detected in 0.4% cases. The clinical picture of Gardnerella infection is similar to that of infection with Mobiluncus, which is little known. This infection occurred 10 times less frequently (2.5%) than gardnerellosis, but in 92% cases it was a component of mixed infection, most frequently in association with G. vaginalis. A little known fungus Leptothrix was found in the genital discharge of 4% examinees, mostly women, but sometimes in men as well; it is represented by 3 types of ramifying threads. The practitioners are to know these infections causing specific diseases, such as bacterial vaginosis, which are often responsible for serious complications in gynecology and obstetrics. PMID- 9410460 TI - [Biovars and antibiotic resistance of enterobacter cloacae strains isolated from colonized newborns]. AB - A total of 120 strains of E. cloacae were isolated from colonized newborns in a maternity hospital (96 strains) and from inpatients of other hospitals (23 strains). Biovars of these strains and of 1 reference strain were determined by two methods of biochemical typing and their sensitivity to 14 antibiotics assessed. The overwhelming majority of isolates from newborns were referred to type 2 according to typing after Old (original) or to biovar 62 if typed using the modification of this method. The phenotypical profile of determinants of resistance to 10 antibacterial drugs were the same, including those to aminoglycosides (except amikacin) and third-generation cephalosporins. The strains isolated from non-obstetrical inpatients belonged to biovars with phenotypical profiles of resistance to only 3 antibiotics and sensitive to aminoglycosides and third-generation cephalosporins. PMID- 9410461 TI - [Diagnosis of chlamydia and gardnerella infection in patients with acute gynecological diseases]. AB - Chlamydial infection was diagnosed by indirect immunofluorescence in 26.3% and Gardnerella infection in 28.7% of patients with acute gynecological diseases. The degree of vaginal contamination in patients with chlamydiasis was virtually the same as in all other examinees, whereas in patients with gardnerellosis the third and fourth degrees of purity predominated. Hence, assessment of the vaginal contamination (purity) is an additional criterion for screening for gardnerellosis but providing no clinically significant information for the detection of chlamydiasis. PMID- 9410462 TI - [Federal system of external evaluation of the quality of clinical laboratory analysis. I. Development of the system]. PMID- 9410463 TI - [Automation in clinical biochemistry (lecture)]. PMID- 9410464 TI - [Available methods of evaluation and correction of immune disorders in patients]. AB - A three-stage scheme for detecting and correcting immunodeficiency is proposed. The first stage is the interview from which the physician may suspect secondary immunodeficiency, the second is assessment of changes in the leukogram, and the third stage is correction of immune disorders with consideration for the immunodeficiency syndrome(s) detected. PMID- 9410465 TI - [Complex immunological examination of patients with allergic diseases]. AB - Allergy is now acknowledged as one form of immunopathology, and the authors propose a protocol of comprehensive immunological examinations for the diagnosis of allergic diseases. This protocol includes the methods of examination and interpretation of the results, permitting the physician detect the predominant mechanism of allergic reaction (reagin, immunocomplex, or cytotoxic) determining the scientifically-based approach to selection of immune therapy. PMID- 9410466 TI - [Legislation of laboratory diagnosis in France]. PMID- 9410467 TI - [Immunoenzyme assay for detection of natural antibodies against catecholamines]. AB - Solid-phase enzyme immunoassay (EIA) for detecting natural antibodies to catecholamines (dopamine, adrenaline, and noradrenaline) has been developed. For this purpose catecholamine antigens were synthesized on polymeric matrix. Optimal conditions for EIA detecting antibodies to these antigens in the sera of donors and patients with schizophrenia and disseminated sclerosis were defined. The levels of antibodies to dopamine, adrenaline, and noradrenaline are constant in donors and shifted in the patients. PMID- 9410468 TI - Sequence comparison of baboon ABO histo-blood group alleles: lesions found in O alleles differ between human and baboon. AB - Histo-blood group O has only rarely been observed in baboon. Recent discovery of such an animal has provided use the opportunity to investigate the molecular genetics of the ABO locus in baboons. The major baboon prototype O allele, observed in two homozygous and several heterozygous animals, is related to the A allele as is the case in humans. Additional apparent prototype O alleles have been observed in heterozygotes, one of which is related to the B allele. The nucleotide changes conferring the O phenotype in the two known human O alleles have not been observed in any baboon allele. This information will aid the identification of baboons useful for the development of xenotransplantation in humans. PMID- 9410470 TI - Compound heterozygotes for hemochromatosis gene mutations: may they help to understand the pathophysiology of the disease? AB - Two mutations have been described on the gene considered to be responsible for genetic hemochromatosis, the HLA-H or HFE gene. The C282Y mutation is a disease causing mutation in most cases of genetic hemochromatosis, but involvment of the H63D substitution in the pathogenesis of the disease is unclear. Compound heterozygotes for both substitutions could help to determine whether or not the second mutation is a worsening factor when associate in trans with the C282Y mutant. We found twenty nine compound heterozygotes during DNA analysis of patients referred to our laboratory for the screening of those mutations. Clinical and biological data were obtainable for 23 of them. Compound heterozygotes could be divided into two groups: subjects with or without iron overload. Five (22%) individuals had normal ferritin levels, whereas 18 had elevated ferritin concentrations (78%). Among those 18 patients, 7 (30% of the total) had clinical and biological criteria of genetic hemochromatosis. Eleven had iron overload without all the criteria of genetic hemochromatosis. Such a high proportion of genetic hemochromatosis is not found in heterozygotes for the C282Y mutation alone neither in our series nor in the literature. Compound heterozygotes for the C282Y and the H63D mutations may have a higher risk of iron overload or genetic hemochromatosis than single heterozygotes for the C282Y mutation. We propose a schematic theoretical representation that could explain this fact at the protein level. Further fundamental studies on the protein, and clinical follow up of compound heterozygotes could help to ascertain this hypothesis. PMID- 9410469 TI - Effect of recombinant interleukin-6 and thrombopoietin on isolated guinea pig bone marrow megakaryocyte protein phosphorylation and proplatelet formation. AB - Guinea pig bone marrow megakaryocytes were isolated and cultured on collagen gels to promote proplatelet formation. In control cultures 15.6% of the cells formed proplatelets. Both IL6 and TPO stimulated dose dependent increases in the percent of proplatelet forming cells up to 26.7% at 100ng/mal IL6 and 26.8% at 100 ng/ml TPO. IL1 and IL3 had no effect on proplatelet formation. IL3 in combination with IL6 and TPO blocked the increase in proplatelet formation observed with IL6 or TPO alone. IL3 was also found to stimulate thymidine incorporation in megakaryocytes. The role of phosphorylation in proplatelet formation was studied using certain inhibitors. The tyrosine kinase inhibitor genestien had no effect on proplatelet formation at concentrations up to 100 microg/ml. The phosphatase inhibitors calyculin A and okadaic acid both inhibited proplatelet formation. Studies on protein phosphorylation revealed that IL6, but not TPO, stimulated phosphorylation of JAK1, JAK2 and MAP kinase. TPO did stimulate tyrosine phosphorylation of Tyk-2. Although IBMX stimulated proplatelet formation, it inhibited phosphorylation of JAK1 and MAP kinase. Adhesion of megakaryocytes to collagen gel also inhibited phosphorylation of JAK1 and JAK2, while MAP kinase phosphorylation was unaffected. These data show that IL6 and TPO stimulate megakaryocyte proplatelet formation. In addition, although these cytokines increase phosphorylation of signal transduction proteins in the JAK/STAT pathway, it appears that a different signal transduction pathway regulated by a combination of phosphatase activity and cAMP levels, leads to proplatelet formation. PMID- 9410471 TI - Impact of HLA-H mutations on iron stores in healthy elderly men and women. AB - The DNA of 287 healthy white elderly volunteers in the New Mexico Aging Process Study, between 63 and 91 years of age, was examined for mutations of the HLA-H gene at nt 845 and nt 187. None were found to be homozygous for the 845A mutation and there were no gender differences in the percentage of the various mutations. The frequency of the 845A mutation was 0.061 resulting in a carrier frequency of 12.2%. The frequency of the 187G mutation was 0.136 resulting in a carrier frequency of 19.9% for a single mutation; 2.4% were compound heterozygous, 834A/187G and 2.4% were homozygous for the 187G mutation. After excluding 5 men and 4 women with microcytic or macrocytic anemia, mean percent transferrin saturation (PSAT) and iron stores, as estimated from serum ferritin concentrations, were calculated for each mutation. Estimated iron stores were normally distributed (range approximately 50 to 1,550 mg) with men (n=111) having significantly higher mean estimated iron stores than women (n=167), 826 +/- 318 and 753 +/- 287 mg, respectively. More men, 15 of 28, (54%) with estimated iron stores in the upper quartile, >/= 1,050 mg, had a HLA-H mutation compared to 25 of 83 (30%) who had a mutation and whose estimated iron stores were < ,050 mg, P<0.05. Seven were heterozygous for the 845A mutation with mean estimated iron stores of 1,300 +/- 127 mg, 7 were heterozygous for the 187G mutation with mean estimated iron stores of 1,439 mg. Similar differences were not noted in women. Even though the potential role of the 187G mutation in the phenotypic expression of HH is less certain than the 845A mutation, the increase in PSAT seen in men with the 187G mutation and the equal distribution of 845A and 187G mutations seen with iron stores >/= 1,050 mg lends support for the involvement of the 187G mutation, or a linked mutation, in iron metabolism. We concluded that men having either a single chromosomal 845A and/or 187G mutation results in higher PSAT's and estimated iron stores than if no HLA-H mutation were present. PMID- 9410472 TI - A high prevalence of HLA-H 845A mutations in hemochromatosis patients and the normal population in eastern England. AB - We have examined normal individuals and all the patients currently being treated for hemochromatosis at the Norfolk and Norwich hospital for mutations in the HLA H gene. We found a gene frequency in 200 normal subjects for teh 845A (C282Y) allele of 0.085, corresponding to a carrier frequency of 17% which is among the highest reported anywhere in the world. The frequency for the less penetrant 187G (H63D) allele was 0.16 among 58 of the normal subjects, which corresponds to a carrier frequency of 32%. All 18 hemochromatosis patients were homozygous for the 845A allele which is not significantly different from other reports in our subset of 12 unrelated patients. These findings present a snapshot of a relatively stable population containing a predicted 3,500 individuals homozygous for the 845A allele but not diagnosed with hemochromatosis. This population will be an excellent model for studies on the penetrance of the 845A homozygous genotype and population screening. PMID- 9410473 TI - Cloning of the murine platelet glycoprotein Ibalpha gene highlighting species specific platelet adhesion. AB - We report the sequence of a 2,779 base pari genomic DNA fragment containing the mouse glycoprotein (GP) Ibalpha gene. Similar to its human counterpart, the mouse GP Ibalpha gene contains a single exon encoding a 734-residue GP Ibalpha precursor polypeptide. Comparative analysis between human and mouse polypeptides reveals a 75% sequence similarity between the amino-terminal domain of each polypeptide. However, there is sequence divergence within a short linear sequence of the amino-terminal domain previously implicated in human GP Ibalpha as critical for the binding of human von Willebrand factor (vWF). Mouse and human primary sequences diverge through their extracytoplasmic macroglycopeptide domains reducing the overall sequence similarity to 70%. The transmembrane and cytoplasmic sequences are highly conserved in both species with 59 identical residues among the 62 comprising the carboxyl-terminus of each polypeptide. The species-specific interaction between human GP Ibalpha and vWF was demonstrated in a model flow system monitoring the ability of surface-bound human vWF to capture from flowing blood normal mouse platelets or transgenic mouse platelets expressing the human GP Ibalpha subunit. The results further define the structural elements necessary for the interaction of human vWF and platelets. PMID- 9410474 TI - Hematologically important mutations: glucose-6-phosphate dehydrogenase. PMID- 9410476 TI - Proceedings of the Vth International Conference on Hormones, Brain, and Behavior. Torino, Italy, August 25-30, 1996. PMID- 9410475 TI - Correlation between genotype and phenotype in hereditary hemochromatosis: analysis of 61 cases. AB - This report assesses the degree of iron overload in a cohort of patients in relationship to the presence or absence of the recently described 845 G-->A (C282Y) and 187 C-->G (H63D) mutations in the HFE (HLA-H) gene. Sixty-one patients with hereditary hemochromatosis diagnosed either with liver biopsy or on clinical grounds were included in this analysis. Forty-one patients were homozygous for C282Y, the genotype considered to be characteristic of hereditary hemochromatosis. At the time of this analysis, 37 of these 41 patients had achieved a state of iron depletion and mobilizable iron was calculated: 19 had less than 4 grams. Twenty-five of these 41 patients had liver biopsies; 4 of these patients had a hepatic iron index less than 1.9. Of the 4 patients with a normal hepatic iron index, 3 had a quantitative hepatic iron of greater than 50 micromol/g dry weight, and one had an inadequate biopsy sample. These findings support our suspicion that individuals may have hereditary hemochromatosis and homozygous C282Y despite relatively low body iron stores. Five patients were compound heterozygotes for C282Y and H63D. Four of these patients underwent liver biopsy; two had a hepatic iron index greater than 1.9. a third patient had a hepatic iron index of 1.3 but a quantitative hepatic iron of 90.6 micromol/g dry weight. All patients were phlebotomized to a state of iron depletion and only one of these patients had a mobilizable iron greater than 4 grams. Three patients were homozygous for H63D; these patients had either a hepatic iron index >1.9 or greater than 4 grams of mobilizable iron. Patients with homozygous H63D and significant iron overload are not well described. Seven patients were heterozygous for either C282Y or H63D; 4 had significant iron overload but three did not. Five patients had no HFE mutations; one of these patients unequivocally has iron overload with a hepatic iron index of 4.4 We conclude that: (1) Identification of HFE mutations will be clinically useful in identifying patients with hereditary hemochromatosis, (2) Patient genotyping will help confirm a diagnosis of hereditary hemochromatosis in some patients with relatively low body iron stores, (3) Significant iron loading can occur in the absence of homozygous C282Y, adding to the evidence that genes other than HFE may be involved in iron loading, and (4) Homozygous H63D can be associated with significant iron overload. PMID- 9410477 TI - Interleukin 5 expressing allergen-specific T-lymphocytes in patients with dust mite sensitization: analysis at a clonal level. PMID- 9410478 TI - Protection against UVA damage and effects on neutrophil-derived reactive oxygen species by beta-carotene. AB - OBJECTIVE: Phagocyte-derived reactive oxygen species (ROS) are involved in microbicidal activities as well as in tissue damage at sites of inflammation. Carotenoids play an important function in protecting cells from oxidant damage. We investigated the in vitro and in vivo effect of 13-cis and 9-cis-beta-carotene on human neutrophils. METHODS: Neutrophils from healthy donors in the presence of 0.25 mumol/L-1 mumol/l beta-carotene and from subjects under beta-carotene supplementation and UVA or UVA/B exposure were stimulated by opsonized zymosan and the generation of ROS was measured by electron spin resonance spectroscopy. RESULTS: Our in vitro results show different effects of the two isomers on stimulated neutrophils. 9-cis-beta-carotene did not produce any change, whereas 13-cis-beta-carotene significantly and concentration-dependent inhibited the ROS generation by stimulated neutrophils. Further, in a controlled study, we were able to demonstrate an in vivo protective effect of beta-carotene on neutrophils against UVA damage by beta-carotene supplemented subjects. PMID- 9410479 TI - [Prevention of childhood disability and medicosocial rehabilitation of disabled children within the framework of a comprehensive program of the Moscow Public Health Committee "Mother and Child Health Protection."]. PMID- 9410480 TI - [Realization of the program "Promotion and support of breast feeding" in Moscow]. PMID- 9410481 TI - [Methods of improving vaccination under conditions of a large city]. PMID- 9410482 TI - [Organization of the substitute therapy service for patients with end-stage chronic renal failure in Moscow]. PMID- 9410483 TI - [Improving the organization of the oncologic service in Moscow]. PMID- 9410484 TI - [Hospice and palliative care]. PMID- 9410485 TI - [Current status and trends in the health of the Moscow population]. AB - Based on vast comprehensive medical statistical database, the authors analyze the health status of the population and the efficacy of public health service in Moscow. The pre-crisis tendencies and the modern status of public health under modern socioeconomic conditions are noted. PMID- 9410486 TI - [Structural changes in the expenditures of public health institutions under conditions of public health reform]. AB - The financial activity of the rural medical institutions in Northern Russia (Arkhangelsk Region) is analyzed. Recent tendencies in alteration of the expenditures of hospitals of different size and level are noted. Basic trends in the process of reformation of public health are outlined, intended to improve the efficacy of utilizing its financial resources which are always insufficient. PMID- 9410487 TI - [Organizational technologies and management of the organs and institutions of the State Sanitary and Epidemiological Surveillance in the Russian Federation]. AB - The authors offer a scientifically-based rationale for the system of management of the State Sanitary and Epidemiological Service and its institutions under modern socioeconomic conditions in Russia. Organizational models of the structure of State Sanitary and Epidemiological Surveillance Centers are offered and their functions determined for various levels of management. A concept of development and organizational structure of the service and its institutions is proposed, aimed at provision of sanitary and epidemiological well-being in the Russian Federation. PMID- 9410488 TI - [Information provision of public health administration at a territorial level]. PMID- 9410489 TI - [Development of obstetric and pediatric care in the Samara region]. AB - The strategic concept of territorial service of maternity and childhood health protection developed in the Samara Region with consideration for obligatory medical insurance can improve the health status of women and children, ameliorate the demographic situation, and decrease the reproductive loss in the Region. PMID- 9410490 TI - [Charter of the Imperial Moscow University of 1804 and introduction of this Charter at the Medical Faculty]. PMID- 9410491 TI - [Medical faculty of Moscow University on the medals of Imperial Russia]. PMID- 9410492 TI - [Development of obstetrics in Moscow in the XIX century]. PMID- 9410493 TI - [Moscow nurses communities in the XIXth and XIIth International Medical Congress in Moscow]. PMID- 9410494 TI - [Problems of public medicine discussed at the XIIth International Medical Congress in Moscow (100th-anniversary of the Congress)]. PMID- 9410495 TI - [The 75th anniversary of the Chair of Social Medicine and Public Health Organization and Economy at the I. M. Sechenov Moscow Medical Academy]. PMID- 9410496 TI - [Zemstvo medicine in the Vyatka province]. PMID- 9410498 TI - Fenfluramine and dexfenfluramine withdrawn from market. PMID- 9410497 TI - [Comprehensive analysis of the health status of the Moscow population]. AB - Several aspects of the Moscow population health are analyzed: quality of life is compared with that in other world megalopolises, medico-demographic values and causes of death in the city are compared with those in other Russian cities, and morbidity in various population groups (children, adolescents, adults) is compared with that in other regions of the Russian Federation. Basic features of the health status of Muscovites are analyzed, as are some negative regularities in the time course of health characteristics, common for the critical period of development of Russia and leading to decrease of birth rate, increase of mortality of capable population, decrease of the expected life span, and increase in the incidence of contagious and other diseases. PMID- 9410499 TI - Parents need help with children's doses of nonprescription drugs, study suggests. PMID- 9410500 TI - Educating parents, providing measuring devices could eliminate pediatric dosage errors, researchers show. PMID- 9410501 TI - Proceedings of the 5th International Conference on Intelligent Systems for Molecular Biology. Halkidiki, Greece, June 21-26, 1997. PMID- 9410502 TI - [Walking disorders and falls in the aged]. PMID- 9410503 TI - [Gait disorders and falls from the neurologic viewpoint. 1. Basic principles: postural control in the aged]. AB - In the framework of gait, disturbances in old age falls of neurologic origin are either due to pareses of the lower extremities or to postural dyscontrol. The latter shows, either alone or in combination, age-associated alterations in its afferent, efferent, and central-integrative components. Considering gait in the elderly as the flexible maintenance of dynamic equilibrium this overview describes the age-associated alterations of postural control. PMID- 9410504 TI - [Gait disorders and falls from the neurologic viewpoint. 2. Clinical aspects]. AB - Any disturbance of gait unmasks itself by a reduced walking velocity. This reduction takes place in order to maintain a low level of energy consumption during walking. The gait phenomenology resulting from velocity reduction is nonspecific and exhibits a compensatory and partly protective walking pattern. Such pattern is intermingled among many neurologic gait abnormalities of old age. Changes in gait patterns or in body equilibrium are among the situations likely to cause a fall in a frail elderly person, particularly, if another disturbance or disease of walking and/or balance control adds to that age-associated pathology. Many neurologic diseases of old age may cause falls by a reduced foot floor clearance during swing phase, thus, raising the risk of stumbling. PMID- 9410505 TI - [Multifactorial pathogenesis of gait disorders, falls and hip fractures in the elderly]. AB - Age, gait disorders, falls, and hip fractures are connected in a pathological cascade. 90% of hip fractures happen at the age of 70 and older; 90% are a consequence of a fall. Only a minority of these patients regain their former level of locomotion. For many of them a hip fracture ends in dependence on personal help, placement in a nursing home, or even death. About 80% of falls occur due to pathological balance and gait disorder and are not due to syncope or overwhelming external force. 5% of all falls result in a fracture, including 1% of all falls with hip fractures. What determines the fracture risk is the relation of bone fragility to geometrical and physical factors of the fall. Regarding the cascade gait disorder-fall-hip fracture, there are always multiple factors interacting. It is useful to give this distinct entity of multifactorial gait disorder with high risk of falling a new diagnostic label: We suggest "age associated multifactorial gait disorder." To identify individual risk factors and regard their interactions can be a basis for therapy and interventions to prevent falls. PMID- 9410506 TI - [Falls in the elderly and drugs]. AB - The risk of falling in older age can be increased by drug use, which has been shown in pharmacoepidemiological studies. There are associations between falls and the use of sedatives, psychotropics, cardiovascular, and other drugs. Multimedication, in particular, has been reported to be frequently related to falls. Mechanisms by which drugs may increase the risk of falling are related to central nervous/neuromuscular and blood pressure lowering effects. Along with increased numbers of diseases and impairments, multiple causes of falls are likely to increase. Thus, it may be difficult to disentangle causal relationships between medication and falls. Disease and impairment related causes of falls always have to be considered. There is still a lack of intervention studies with special regard to medication although drug prescribing is given a high priority in prevention of falls in older age. In medical practice, adaptation of dosing and additive pharmacodynamic effects of multiple drugs should be carefully considered. PMID- 9410507 TI - [Tinetti motor ability test: sensitivity to change in gait assessment during geriatric hospitalization--aspects of its clinical relevance and quality assurance]. AB - Assessment instruments are recommended in addition to the clinical examination of gait disorders in elderly patients. We examined the sensitivity of gait assessment in a geriatric hospital by using a modified Tinetti's motility score in order to study aspects of clinical relevance and quality assurance. Forty patients were assessed on admission and discharge. The results were rated for information profit in comparison to clinical admission report. Three geriatricians rated the results for clinical relevance. Three quarters of the patients' mobility changed significantly on the course. There were no significant differences in patients with dementia. Assessment by using Tinetti's motility score gained 41% new information and 49% partly new information. This information was rated as considerably relevant to the clinical course in 56% to 80% of the patients. A structured motility score is useful for the purpose of internal quality assurance in a geriatric hospital. PMID- 9410509 TI - [Effectiveness of multi-factorial intervention for reducing falls with proximal femoral fractures in homes for the aged and nursing homes. Goals and study design of a population-based study]. AB - The article presents the design of an open controlled, non-randomized population based intervention trial in institutionalized elderly that will be performed in Ulm/Germany starting in 1998. The multimodal intervention includes education, counseling, training, and hip protectors as well as information on potential environmental modifications in nursing homes. Two historical and one geographical control group will be formed. All institutionalized elderly will be assessed by means of the Minimum Data Set of the Resident Assessment Instrument. The intervention will last 12 months. The main objective is to test a model for cost effective reduction of proximal femoral fractures in institutionalized elderly and to demonstrate its effectiveness. PMID- 9410508 TI - [Strength and coordination training for prevention of falls in the elderly]. AB - Since muscle strength and coordination training in healthy elderly people has been effective in the primary prevention, a program for geriatric patients with falls is presented. Its aim is safer walking by an increase in muscular strength and improved coordination. The exclusion criteria and the size of the training program define a group of over 75 year old patients, who will be compared with a group with light physical exercise in a randomized fashion. The first results show a remarkable increase in strength and a low complication rate. The functional relevance of our program remains to be proven. PMID- 9410511 TI - [Value of exercise and sports activities in the elderly]. AB - The sport scientific research within the scope of the first survey of the ILSE longitudinal study consisted of standardized interviews of 695 probands born between 1930 and 1932. Based upon these interviews will first be presented the connections between physical activities on the one hand and the subjective estimation of health and physical ability on the other. Then a comparison between these subjective estimations and the objective medical opinions will follow. Physical activity has a significantly positive influence on both the subjective estimations and the general attitude towards age and contentment with life. Furthermore the examination provided information on the expectations the elderly link with physical activities, why they possibly refuse to exercise, and under which conditions they might start to exercise. The most important result is the main position of health for the decision in favor of or against physical activities. The statements of the physically active on the exercised sports provide indications of the different interests of men and women and the desired offers. PMID- 9410510 TI - [Expanded therapeutic possibilities in hyperthyroidism in the elderly. Sclerotherapy of autonomous adenomas and thyrostatic therapy]. AB - Patients with Graves' or Plummer's disease have usually definitively been treated by surgery or radioiodine. If, however, the patient does not accept either form of treatment or the risk of surgery or the inconveniences of the radioiodine treatment by far out-weight the expected benefit for the patient, alternative therapeutic approaches should be explained and been offered to the patient: 1) It must be clear, whether any treatment is necessary at all: Small compensated autonomous adenomas with euthyroidism must not be eliminated in every case, the same applies for cases of remaining subclinical hyperthyroidism after radioiodide or surgery if the amount of remaining functional tissue is small. 2) Isolated autonomous adenomas may, under certain conditions, be eliminated by alcohol injections. This novel approach should be considered particularly for elderly patients, since it can be done on an outpatient basis. 3) For very old and very frail patients a continuous and lifelong medical treatment represents and adequate alternative to surgery and radioiodine. Small amounts of carbimazole or methimazole in a single daily dose combined with small amounts of thyroxine may safely control the hyperthyroidism, again on outpatient conditions. PMID- 9410512 TI - [The "restraint" controversy in geriatric psychiatry. Focus study of factors influencing restraint measures]. AB - The study examines the relationship between frequency and duration of restraints in psychogeriatric inpatients with their respective diagnosis, medication, reason for restraints, and age as a socio-demographic variable. Entered in the analysis were data from 590 inpatients from a total of 29 institutions within the Federal Republic of Germany psychogeriatric facilities; this was 24.7% of all psychogeriatrically treated inpatients on the survey day in the said 29 institutions. The data were analyzed via simple multifactorial analyses of variance followed by multiple classification analysis (SPSS for Windows). Neither age nor diagnosis showed any differential influence on frequency or duration of restraints. Among the reasons for restrains hetero-aggression led to fewer and shorter restraints; self-aggression led to fewer but longer restraints. Restless patients were restrained more often but for shorter intervals. The main psychopharmacological strategy, while having no influence on the frequency of restraints, showed a marked influence on their duration; patients treated with low potent neuroleptics had particularly short intervals of restraints, whereas patients free from psychopharmacological agents showed distinctly longer intervals of restraints than the mean of restrained patients. PMID- 9410513 TI - [Comment on the contribution by I Reich, V. Herrwerth, I. Marintschev, L. Gebert (1997). Physical capacity and mortality of elderly patients after surgical management of para-articular hip fractures]. PMID- 9410514 TI - [Prenatal guidelines and Chlamydia screening]. PMID- 9410515 TI - [Prenatal toxoplasmosis--do we need screening in pregnancy?]. PMID- 9410516 TI - [Results of follow-up of mothers with previous surgical "total cervical cerclage" also with reference to neonatal data]. AB - From August 16, 1993 until March 6, 1995 a study was conducted at the Department of Obstetrics at the Berlin-Neukoelln Womens Hospital to evaluate the status of patients who had previously undergone total cervical occlusion (TCO) for the prevention of late abortion and premature birth. Special consideration was given to the possible effects of TCO on the patients' gynecologic status, the psychological circumstances associated with TCO and the developmental status of the infants. Fifty-four women participated in the study, their ages ranging between 26 and 53 years. The mean time-span between the TCO procedure and the present study was five years and five months. In the 54 women undergoing TCO, a total of 73 such procedures were performed. An extensive early TCO was performed in 63% and an extensive late TCO in 20.5%. The gestational age at the time of delivery was > or = 37 + 0 weeks in 67.1% of the women. More than half of the patients (54.9%) experienced a normal spontaneous vaginal delivery. In total, 84.5% of the infants were delivered vaginally; thus the cesarean section rate was only 15.5%. Regarding the patients' medical histories, the majority of the patients had no pathological findings on routine pap smears and pelvic examinations and, similarly, they had no complaints of menstrual irregularities. Only 10 patients (18.5%) underwent gynecological surgery in the interim; in these patients, the most common reason for surgery (in four cases) was sterilisation. The findings during speculum examination were tabulated. The majority of the patients (96%) exhibited a normal multiparous cervical portio. In 55% of the patients there was no evidence of scarring of the cervical portio. In 25% of the patients there was minor cervical scarring, in 13.5% it was moderate and in 5.8% it was severe. Except for a single case, the patients showed no evidence of vaginitis. During bimanual palpation on pelvic examination, in 82.7% of the patients the cervix was found to be at least 2 cm in length, a closed external cervical os was palpated in 65.4%, and an anteflexed/anteverted uterus that was normal in size and form was noted in 46.2%. In 95.4% of the patients, original squamous epithelium was seen colposcopically. The psychosocial status of each patient was evaluated on an individual basis. After taking all of the psychosocial circumstances associated with such a high-risk pregnancy into consideration, all the patients giving birth to a living infant described that event as a positive experience. However, this was not the case in the two patients whose premature infants did not survive. In 74.1% of the patients, the relationship with their domestic partners was described as "unchanged" when compared to the status of their relationship during the preceding pregnancy without TCO. The patients undergoing TCO described their own psychological status as "frequently strained" (61.1%) and "disturbed" (9.3%) antepartum. Postpartum, 90.7% of the patients described their psychological status as "good". In total, there were 74 births in the 54 patients included in this follow-up study. Of these 51 (68.9%) were living term infants, one stillborn (1.4%) and 22 (29.7%) premature infants. One infant weighed less than 1.000 grams while 4 (5.3%) weighed between 1.000 and 1.499 grams. Postpartum, 45.5% of the premature infants and 19.6% of the term infants were hospitalized. The primary indications for the transfer of the premature neonates were intensive care, hyperbilirubinemia and adaptation problems. The physical, emotional and mental development was unimpaired in 91.4% of the children. A capability for good social integration was displayed in 95.7% of the children. Based on the current investigation, it may be concluded that the total cervical occlusion procedure has no significant long term negative effects. PMID- 9410517 TI - [What effect does leg position in breech presentation have on mode of delivery and early neonatal morbidity?]. AB - The objective of the present study is to establish whether the position of the legs in breech presentation deliveries affects the vaginal or abdominal mode of surgical delivery and the early neonatal morbidity. The patient population investigated (n = 266) comprised 163 primiparae (61.3%) and 103 multiparae (38.7%). Of the 266 term infants (more than 37 complete WOP) with breech presentation, 71.3% (127/178) could be delivered vaginally from a simple pelvic presentation and 55.3% (42/76) from an incomplete or complete footling presentation. The average duration of labor was 460 minutes in a pelvic presentation delivery, and 400 minutes in a footling presentation delivery. The rate of acidosis (pHNA < 7.20) was 26% in the neonates delivered from the pelvic presentation as compared to 11.9% in the footling presentation deliveries. The number of neonates with a 5/10 minute APGAR score of seven points was 0/0% in pelvic and 4.8/2.4% in footling presentation. Of the neonates delivered from pelvic presentation, 10.2% (13/127) were moved to the neonatology department as compared to 14.3% (6/42) babies delivered from footling presentation. A secondary Cesarean section was indicated in 28.7% of pelvic presentation deliveries commenced vaginally (51/178) and in 44.7% of the footling deliveries (34/76). The rate of acidosis was 49% (25/51) in the babies with pelvic presentation and 21% (7/34) in the babies with footling presentation. The percentage of neonates with respiratory depression (5/10 minute APGAR score < 7 points) was 3.9/2.9% in the babies delivered from pelvic presentation and 2/0% in the babies delivered from footling presentation. Correspondingly, the rate of transfer to the neonatology ward was extremely high: 33.3% (17/51) of the pelvic presentation babies and 8.8% (3/34) of footling babies. In 11 pregnant women (5.8%) with a fetus in pelvic presentation, a primary Cesarean section was indicated, in half of these cases (n = 5) because of "suspicion of a discrepancy", three times at request of the patients and three based on fetal and on maternal indication. Because she had rejected vaginal delivery, primary Cesarean section was performed in one pregnant mother (1.3%) with a fetus in footling presentation. Fetuses with simple pelvic presentation at term were more frequently delivered vaginally than fetuses in footling presentation (71.3% compared to 55.3%). The rate of respiratory acidosis was higher in the neonates with pelvic presentation than those with footling presentation (26% as compared to 11.9%). They compensated this acidotic metabolism within a short time, so that the transfer to the neonatology ward was only temporary. The same applied to the babies delivered by secondary Cesarean section. PMID- 9410518 TI - [Effect of Doppler ultrasound on perinatal mortality of high risk pregnancies: required patient sample?]. AB - The clinical value of Doppler ultrasound in obstetrics is related to the question whether pregnancy course and outcome will be improved when this method is introduced into obstetrical management. Recently a couple of randomized controlled management studies have been published and the majority of these trials showed significant beneficial effects. Even perinatal mortality may possibly be reduced, but none of these trials was large enough to achieve a significant reduction. Giles and Bisits [12] performed a metaanalysis by combining evidence from 6 of these trials in high risk pregnancies and could demonstrate a 50% reduction in perinatal mortality at a significance of 5%. But they did not report whether the sample size was large enough to detect this difference with the usually required power of at least 80%. Therefore the question remains whether obstetrical management should be changed in view of these results. Additionally, a clinically useful estimate is needed to determine the chance to reproduce the assumed beneficial effect. The present paper describes the statistical method for calculating the appropriate sample size needed in each group of a trial to detect a predefined treatment difference with specified significance and power. This method is applied in detail to the results of the meta-analysis of Giles and Bisits [12] and a minimal sample size of 1458 high risk pregnancies in each group is calculated. As the Doppler and the control groups actually contained 2102 and 2133 high risk patients respectively, these sample sizes proved to be large enough to detect a 50% reduction in perinatal mortality with a power of at least 80%. Furthermore, 59 high risk pregnancies would need to be managed by Doppler in order to prevent 1 case of perinatal death. In other words, it should be possible to prevent 4 perinatal death cases per 1000 total births when the high risk proportion is 25%. PMID- 9410519 TI - [Induced labor with prostaglandins in birth weight below 2,500 g]. AB - In a multicenter study the deliveries of 127 newborn infants with a birth weight less than 2500 grams were evaluated after induction of labour using prostaglandins, among them 90 infants with a birth weight below the 10. percentile in relation to the gestational age. The control group were 1566 deliveries of newborns with normal weight, delivered at term after induction with prostaglandins. The rate of vaginal deliveries was 67.6% in the group of newborns weighing less than 2500 grams and 84.1% in the control group. The rate of acidotic newborns (pH < 7.20) was remarkably higher in newborns below 2500 grams (12.6%), especially in the subgroup of growth retarded newborns (16.7%), when compared to the control group (9.5%). In primaiparae with fetal growth retardation and low Bishop score the induction of labour with prostaglandins resulted in an insufficient progress of labour. In this subgroup the rate of cesarean deliveries was 83.3%. Induction of labour was substantially more successful after premature rupture of membranes. The most favourable results were achieved in multiparae after premature rupture of membranes which resulted in vaginal deliveries in 94.5% of cases independent of gestational age. PMID- 9410520 TI - [Delivery following surgically treated sacrococcygeal teratoma in the mother]. AB - The delivery of a 23 para I who was born herself with a sacrococcygeal teratoma of the type II is reported. After the extirpation of the tumor and of the coccyx an anomaly of the pelvis is found in the sense of a pygopagic pelvis (stretching and bending of the sacrum towards the dorsum). This anomaly and the scars in the way of a maternal soft tissue dystocia seem to be the reasons for the insufficient progression of labour. The delivery had to be done by caesarean section. A historical review concerning the surgery of sacrococcygeal teratomas is given. PMID- 9410521 TI - [Breast feeding recommendations by the German National Breast Feeding Committee]. PMID- 9410522 TI - [Comment on S. Bussen, G. Schwarzmann, T. Steck: Clinical aspects and therapy of amniotic fluid embolism. Presentation of a case]. PMID- 9410523 TI - [Physiology of the placenta: development of fetal nutrition during pregnancy]. AB - In addition to placental growth during the first half of pregnancy, remodelling of the placenta which takes place in the second half of pregnancy contributes significantly to the increase in transport capacity in late pregnancy. Due to the exponential growth of the fetus the balance between supply and demand is shifted towards increasing demand and fetal blood concentrations of important substrates like glucose and oxygen decline. The decrease in fetal blood levels of glucose and oxygen resulting in an increased transplacental gradient add to a rise in flux from the mother to the fetus. The change in distribution of the total uptake from maternal blood between placenta and fetus allows a further increase in the amount of substrate which is available to the fetus and helps to balance the equilibrium between supply and demand. Disturbances in growth and development of the placenta in early pregnancy lead to an earlier and more pronounced manifestation of the compensatory-mechanisms supporting supply of the fetus, which normally are only seen in late pregnancy. An increase in the ratio of fetal to placental weight is an indicator of the compensatory mechanisms. PMID- 9410524 TI - [Pregnancy and sports--physiologic considerations and practical examples]. AB - Mothers-to-be expect practical and competent consulting from their obstetricians rather than orders or interdictions regarding their way of living. Particularly answering questions concerning physical activity and sports during pregnancy require profound knowledge on the physiological adaptations of the cardiopulmonary system and the anatomical structures and-on the other hand-on performance and sports' physiology. Resuming these alterations through pregnancy there result practical examples and proposals of physical activities in gestation as therapeutic and preventive measures. Moreover risks and contraindications are worked out. PMID- 9410525 TI - [Micronutrients in pregnancy]. AB - During pregnancy the demands for energy and far more for micronutrients are increased. Discrepancies between the intake of certain micronutrients such as iodine, iron and folic acid and the recommended quantities during pregnancy are to be expected. Consequences of an unsufficient supply of these micronutrients might be goitre, anemia and with special reference to folic acid the occurrence of neural tube defects. Deficiency can be prevented by careful choice of food. Supplementation with special micronutrients is necessary before respective during pregnancy however for example to prevent neural tube defects by periconceptional supplementation with folic acid. PMID- 9410526 TI - [Tocolysis]. AB - Effectiveness of tocolytic strategies are difficult to prove and should called in question. Tocolysis by betaadrenergic agents inhibit a symptom without eliminating causing pathologies. Initial steps in etiology as well as therapy should be based on the entire organism including the psyche with a special focus on uterus, cervix and feto-placental unit. Magnesium depletion and infection are important causes of preterm labour. Application of betaadrenergic agonists should be pulsatile by bolus tocolysis demonstrating rare side effects. PMID- 9410527 TI - [Criteria for successful outcome of external fetal version from breech presentation to cephalic presentation]. AB - The technique of external version of breech presentations has been proposed as a tool to reduce morbidity of fetus and mother. 79 cases of a 3 year period were evaluated aiming at identification of risk factors improving the success rate of the intervention. A total of 48% of attempts was successful. The most frustrating single factor was identified to be the oligohydramnion. Posterior implantation of the placenta improved the rate to 61%. The umbilical cord being localized by coloured Doppler had little influence even if positioned around the neck as neither success nor complications were predictable in this case. Maternal adipositas had a negative influence on the success rate. Perinatal morbidity war not increased in the group of external versions. PMID- 9410528 TI - [Deciding criteria for vaginal delivery from breech presentation--effect on neonatal early and late morbidity]. AB - In the past 20 years, breech presentation obstetrics has been characterised mainly by the work of Kubli (1975): "The safest and most simple way to prevent the foetal obstetric risk of breech presentation is the systematic Caesarean section." In the past, this resulted in judicial decisions in which due to vaginal delivery a child has been damaged and the doctor has been reproached for not performing a Caesarean section. In the report of the "Standardkommission BEL" (1983) as well as in the Guidelines for the Management of Breech Delivery (FIGO 1993), subtile criteria of decision were demanded for the vaginal delivery method. To an increasing extent clinics lack the necessary training for vaginal operative delivery for breech presentation, followed by economic obligations and, as a result, in 1994 the frequency of Caesarean sections in cases of breech presentation was 86.9% in Bavaria. Investigations should be made as to whether it is actually necessary, for the benefit of the child, to subject a mother of a breech baby to such high-risk operations. From 1988 to 1995, 1116 breech babies were born in the Gynaecological Clinic Nurnberg, Dept. of Obstetrics. The case history of 650 children with a period of gestation > 32 weeks of pregnancy were evaluated: parity, period of gestation, method of delivery, condition after Caesarean section, premature rupture of the foetal membranes, position of legs, weight of child, Apgar score, umbilical artery pH, base excess in umbilical artery mval/l, transfer ratio to paediatric clinic, reason of transfer to paediatric clinic, rate and duration of intubation, cranial sonogram and electroencephalogram. PMID- 9410529 TI - [Interpretation of pulse oximetry and near infrared spectroscopy values sub partu]. AB - Pulse oximetry and Near Infrared Spectroscopy (NIRS) are both optical methods which have been made applicable to the fetus during labour in order to improve the assessment of fetal oxygen supply and well being. 317 pulse oximetry measurements showed that postpartum pH, Base Excess, PO2 are significantly negatively resp. pCO2 is positively correlated to the time, during which arterial oxygen saturation (SpO2) is below a critical threshold of 30%. The 1' APGAR-score is also influenced by the duration of SpO2 values below 30%. Near Infrared Spectroscopy semiquantitatively measures cerebral concentration changes of oxygenized, desoxygenized and total hemoglobin as well as that of cytochrome aa3 which is the key enzyme of the intracellular respiratory chain. In an analysis of 1350 contractions during 42 deliveries decelerations type 1 and 2 differed from normal CTG by a significantly deeper drop of oxygenized and total hemoglobin in the brain on top of contractions. Until the following contraction oxygenized and total hemoglobin remained deeper below their original levels in case of decelerations type 2 than in case of normal CTG or type 1 decelerations. Simultaneous registration of pulse oximetry and NIRS revealed an increase of cerebral blood volume in case of low arterial oxygen saturation (SpO2 < 30%) which may be interpreted as "brain sparing effect". PMID- 9410530 TI - [Episiotomy and its complications]. AB - 1. Many benefits claimed for episiotomy are not sufficiently proven. In recent literature, some of them are questioned and some have been disproven. 2. Episiotomy, especially median episiotomy, has a higher risk of third-degree lacerations. Mediolateral episiotomy is more often followed by postpartum pain and impaired wound-healing. 3. Typical, albeit rare complications of episiotomy and third-degree lacerations are incontinence for stool and flatus, and-very seldom-fistula formation. 4. Complications of episiotomy as well as the failure to perform an episiotomy have had forensic consequences. 5. For good healing of an episiotomy or a perineal laceration suturing with an adequate technique and the use of non-reactive suture material is mandatory. PMID- 9410531 TI - [Premature labor, determination of fetal lung maturity and medicamentous induction of lung maturation]. AB - Preterm delivery continues to be one of the main causes of perinatal mortality and morbidity, comprising about 8% of all deliveries. The percentage of the most jeopardized preterm newborns weighing less than 1500 g is about 1, however, can be several fold higher in perinatal centers. Respiratory distress syndrome, intracranial hemorrhage, persistent fetal circulation as well as enterocolitis represent prominent complications. Avoiding preterm delivery in cases of preterm labor is attempted by application of magnesium sulfate, beta sympathomimetics or non-steroidal antiphlogistics. Is preterm delivery desirable, however not absolutely indicated, knowing the status of the fetal lung maturity may be helpful in the decision making process. This can nowadays best be achieved by determining the surfactant albumin-ratio by possibly repeatedly withdrawing amniotic fluid by amniocentesis. When inducing lung maturation, application of corticosteroids crossing the placenta is favoured. With impending preterm delivery this should always be given a try, independent from the time expected to be left until delivery- and also below the completed 27th week of pregnancy. Prenatal induction of lung maturation and postnatal application of surfactant are to be looked at as complementing therapeutic concepts in the immature neonate. PMID- 9410532 TI - [The role of extracorporeal membrane oxygenation (ECMO) in current neonatology]. AB - Two hundred term or near-term neonates were referred to an ECMO center for severe PPHN associated diseases. In 2 time periods from 1987 to 1991 and from 1992 to December 1995 alternative treatment modes were tried in an attempt to obviate ECMO. During the first time period patients underwent a trial of high-frequency oscillatory (HFOV) ventilation before ECMO. In the second time period patients first received inhaled NO followed by HFOV in non-responders. If this also failed HFOV was combined with iNO. In both time periods about 40% of the patients were spared ECMO treatment by these alternative treatment modalities. INO only benefited 15% of the ECMO candidates who apparently had fared just as well on HFOV alone in the preceding time period. While patients who were improved by iNO were spared HFOV with its potential severe complications, i.e. air leaks and cardiocirculatory instability. More extended long-term studies will have to show which of these 2 treatment modalities (iNO or HFOV) should be given-first priority in an attempt to avoid ECMO in neonates with severe respiratory failure. PMID- 9410533 TI - [Modification of transplacental digoxin transfer in the isolated placental lobule]. AB - Digoxin is widely used in the transplacental therapy of fetal tachyarrhythmia. Unfortunately, in cases with severe cardiac insufficiency and hydrops fetalis, transplacental passage of digoxin is often hampered and therapy therefore ineffective. The present study was designed to establish the isolated placental lobule to quantify transplacental digoxin passage under different experimental conditions. Ten human placentas were obtained immediately after delivery, and a lobule was dually perfused after cannulating a small artery and vein of the chorionic plate and piercing four catheters through the corresponding basal plate. Flow rates were 12 ml/min in the maternal circuit and 6 (I) respectively 3 ml/min (II) in the fetal circuit. The maternal circuit was spiked with digoxin to 6.18 +/- 0.40 ng/ml, and transplacental passage was calculated from repeated fetal and maternal perfusate samples (Fluorescence-Polarization-Immunoassay; TDx, Abbott Laboratories). Within three hours of recirculating perfusion with a fetal flow rate of 6 ml/min (I), digoxin concentrations in the maternal circuit (400 ml) declined to 3.56 +/- 0.09 ng/ml, whereas digoxin levels in the fetal compartment (200 ml) increased to 2.58 +/- 0.37 ng/ml. With a fetal perfusion rate of 3 ml/min (II), the efflux of digoxin out of the maternal circuit was lower (p < 0.05) and the influx in the total compartment was reduced (fetal digoxin concentrations reached only 26.9 +/- 10.6% vs. 39.1 +/- 5.5% of the initial maternal digoxin concentrations). These data suggest that severe fetal cardiac insufficiency with reduced placental perfusion may be in part responsible for the decrease of transplacental digoxin passage in fetuses with hydrops. PMID- 9410535 TI - [Analysis of the Doppler signal (color and pulsed) in hepatocarcinoma]. AB - Real time echocardiography associated with pulsed Doppler (duplex) and color (triplex) provide a non invasive technique to determine the vascular morphology of a lesion. The duplex and triplex signal of 35 liver tumors, 23 hepatocarcinomas and 12 angiomas found in the first high frequency signals greater than 1.81 kHz and traced with pulsatile or mixed morphology (pulsed and continuous) were analyzed. The second signals were of low frequency, less than 1 kHz and continuous tracing. It may be concluded than echo-Doppler is a complementary technique to conventional echography and is useful in the diagnosis of hepatocellular carcinoma. PMID- 9410534 TI - 3rd European Conference on Gene Therapy of Cancer. Berlin, Germany, September 11 13, 1997. Abstracts. PMID- 9410536 TI - [Immunohistochemical localization of annexin VI in the endocytic compartment of rat liver hepatocytes]. AB - Annexin VI has been isolated from rat liver endosomes and affinity purified antibodies have been produced. By Western blotting, in rat liver subcellular fractions, anti-annexin VI was demonstrated to recognise a 68 kDa band in the three endosomal fractions. In the present study, immunogold labeling of ultrathin Lowicryl sections of rat liver has been used to get insights into the ultrastructural hepatocyte localization. Although at the immunofluorescence level the staining seemed located at the apical, canalicular plasma membrane, domain of the hepatocytes, the electron microscopy revealed that 80% of the labeling, with the anti-annexin VI antibody was specifically localized not at the plasma membrane but in the close subapical endocytic compartment surrounding the bile canalicular plasma membrane of the hepatocyte. Double immunogold labeling with an anti peptide antibody to Rab5 and anti-annexin VI showed that 80% of the Rab5 positive apical endosomes were also labeled with anti-annexin VI antibodies. However, there was no significant colocalization of annexin VI and structures labeled with antibodies to the polymeric immunoglobulin receptor. The results suggest that annexin VI could be involved in regulating the functioning of this apical compartment in the hepatocyte. PMID- 9410537 TI - [Prevalence of cholelithiasis in El Real-Gandia]. AB - The aim of this study was to know the prevalence of cholelithiasis (C) in the El Real-Gandia (Spain) as well as the degree of the response of the population. Health examinations were performed in 1,803 adults from El Real (2,000 inhabitants) and Gandia (54,000 inhabitants) using abdominal echography as the screening technique. Cholelithiasis was defined as the presence of biliary lithiasis (BL) or previous cholecystectomy (PC). Of 1,268 (70.3%) participants in the study, C was found in 126 cases (BL in 102 and P in 24) representing a standardized prevalence of around 15% in women and 5% in men. Cholelithiasis was more frequent in females (13.8%) than in males (5.7%) (p < 0.001) increasing linearly with age (p < 0.005). The proportion of PC was significantly higher in women (23.9%) than in men (5.9%) (p < 0.05) and in Gandia (34.4%) than in El Real (13.8%) (p < 0.02). The prevalence of biliary mud and polyps was of 0.3% and 1.1%, respectively. Working obligations (35.4%) and fear of hospitals (22.4%) were the most frequent causes for no response (NR). Males with more than primary a school education originating from outside the Valencian community (VC) were significantly associated with NR in multivariant analysis. PMID- 9410538 TI - [Multifocal Langerhans-cell granulomatosis with hepatic lesions simulating tumors]. AB - Langerhans cells granulomatosis is a rare disease characterized by the clonal proliferation of the Langerhans cell, a cell element pertaining to the mononuclear phagocytes system. Hepatic involvement may be presented, particularly in the multifocal or disseminated form, together with the remaining surrounding organs. Radiologic findings have recently been reported including echographic, computerized tomography and magnetic resonance of the hepatic lesion of the disease. The case of a patient whose initial radiographic study suggested the existence of hepatic metastasis and which was later diagnosed with multifocal granulomatosis of Langerhans cells with hepatic involvement is reported. Radiologic and histologic images are provided and the data reported in the literature concerning this disease are reviewed. PMID- 9410539 TI - [Psoas abscess in Crohn's disease. Preoperative evaluation and surgical attitude]. AB - Three patients with psoas abscess secondary to Crohn's diseases are reported with the aim of discussing the diagnosis and surgical management of these patients. It is concluded that computerized tomography is the method of choice for the diagnosis and follow up of this complication. Surgery should be individualized but in stable patients abscess drainage should be attempted in addition to resection of the intestine involved. Finally, given the great risk of pulmonary thromboembolism in these patients, intensive prophylactic anticoagulant therapy is required. PMID- 9410540 TI - [Massive pulmonary calcinosis and soft tissue calcifications in liver transplant. Presentation of a case associated with severe cytomegalovirus infection]. PMID- 9410541 TI - [Alagille syndrome and isolated persistent ductus arteriosus]. AB - The clinical case of a 21-year-old patient diagnosed with a variant of intrahepatic biliary ductopenia or Alagille syndrome, a dominant inherited disorder associated with a chronic cholestatic syndrome and abnormalities in different organs and systems. The present article discusses the main clinical abnormalities found in this syndrome and we describe a persistent arterial ductus as an isolated cardiovascular manifestation not previously reported in association to this syndrome. PMID- 9410542 TI - [Lamivudine treatment: realities and expectations]. PMID- 9410543 TI - [Reservoiritis: acute inflammation of the ileoanal reservoir (I). Etiopathogenesis and diagnostic criteria]. PMID- 9410544 TI - [Reservoiritis: acute inflammation of the ileoanal reservoir (and II). Natural history and treatment]. PMID- 9410545 TI - [Malaria brought into Strasbourg: an epidemiological, clinical, biological and therapeutic study]. AB - In a retrospective study, we registered 210 patients hospitalized in Strasbourg for malaria from 1984 to 1995. The diagnosis was always confirmed by presence of the parasite on blood smears. We analysed the epidemiological, clinical, biological and therapeutic data. The number of cases rose each year, with a maximum in 1995. The majority of cases occurred in January and from August to October, these periods corresponding with the return of travelers. In most cases, infection took place in Africa. In this region, Plasmodium falciparum is the most frequent species of the parasite. The mean age of the patients was 33 years. The clinical manifestations were polymorphic: fever, chills, sweating, and headache were very frequent. We noted 15 serious infections (with the WHO's definition) and two cases of cerebral malaria. All cases had a favorable outcome. Five cases occurred in pregnant women; two of them had a severe form of malaria. Among the biological abnormalities, we found thrombopenia, haemolysis, hypocholesterolaemia and hypertriglyceridaemia. The significance of disturbance of the lipid metabolism is not known. Hypocholesterolaemia is very frequent, and hypertriglyceridaemia seems to be associated with severe malaria. Most malaria attacks occurred in patients without adequate chemoprophylaxis. This confirms the importance of prophylactic information given to patients by their physicians. Resistance develops against each new medication that is available; among these quinine remains the drug of choice to cure severe malaria. PMID- 9410546 TI - Associateships of a different kind. PMID- 9410548 TI - Targeted chromosome breakage in filamentous fungi. PMID- 9410547 TI - [Treatment cost savings with universal coverage of dental pit and fissure sealants in Quebec]. AB - Approximately 95 per cent of caries or fillings in the first permanent molars of eight-year-old children are found on pit and fissure surfaces. In 17-year-old adolescents, these account for 68 per cent. This article evaluates two treatment approaches based on their respective cost: one that does not use sealants and one that uses sealants for pit and fissure surfaces and amalgam fillings for restorations involving other surfaces. Compared to the current situation in which sealants are rarely used, applying dental sealant to three out of four first permanent molars reduces the cost of treatment by 31 per cent. Therefore, pit and fissure sealants are recommended as a universal preventive measure for children. In the long run, this could generate savings of up to $7 million in the public and private sectors for each age group in Quebec. PMID- 9410549 TI - [Psychometric evaluation of an instrument measuring motivation]. AB - McEwen's Health Motivation Assessment Inventory (HMAI) (1993) tool was developed to assess the motivation of patients with coronary artery disease to initiate and sustain healthy habits. Because of its measurement difficulties, it was modified and translated into French. The purpose of this methodological study was to examine the psychometric properties (content validity, internal consistency, and test-retest reliability) of the modified HMAI on 255 normal subjects. The average proportions of the items, rated congruent by the three raters, were 0.99 for clarity and 0.95 for relevancy. The final Cronbach's alpha coefficients for the six subscales ranged from 0.08 to 0.67. Confirmatory factor analysis provided evidence that the majority of the items were moderately independent of each other. Also, temporal stability coefficients ranged from 0.49 to 0.81. Finally, the modified HMAI was found to be free of social desirability bias. Although the modified HMAI appears to be a promising tool for future research, further refinement is needed to improve its validity. PMID- 9410550 TI - [Enterococci in human environment]. AB - Enterococci, formerly confounded with faecal streptococci, are recognized since the beginning of the century as being faecal in origin and are generally searched for in waste waters and food products; their detection may in fact indicate the presence of enteropathogenic organisms. Although nearly ubiquitous, their preferred ecological niche is the intestine sphere. Rejected in the environment by means of human faeces or animal dejecta, they are scattered afterwards in diverse niches. Once in the external environment, their survival is linked with their exceptional aptitude to resist or grow in hostile environments that are usually detrimental to the development of most mesophilic microorganisms. However, a certain ambiguity exists concerning their relationships with human beings. In fact, certain enterococcus strains or species are used in the elaboration of some milk products. Conversely, others are opportunists and may cause severe infections to people from infants to adults. Moreover, undergoing adaptation perpetually, they present a multiresistance pattern to antibiotics. Thus, the barrier that separates bacteria as nonoffensive contaminants from powerful pathogens appears most fragile, suggesting that people must systematically consider suspect the presence of enterococci in their near environment. PMID- 9410551 TI - [Insulin therapy for non-insulin-dependent diabetes: minimal or intensive]. AB - The indications for continuous insulin therapy during non-insulin-dependent diabetes (NIDD) are numerous. In addition to patients with a contraindication to oral treatment, the greatest cause is "failures resulting from the use of hypoglycaemic agents". According to data in large series published to date, these secondary failures occur at an annual rate ranging from 2 to 10% and are more frequent in subjects whose weight is normal or moderately high. In current medical practice in France, the indications for insulin therapy are considered late, in the presence of severe hyperglycaemia indicative of beta-cell failure. From then on, the problem raised is that of the glycaemic goal to be reached, which has an influence on the therapeutic strategy to be adopted. In addition to the risk of microangiopathy, NIDD patients run a very high risk of macroangiopathy, particularly when insulin therapy is initiated late. In patients whose life expectancy is fairly long (7 to 10 years or more), a body of convergent clinical and epidemiological evidence favours strict glycaemic control, i.e. intensive insulin therapy. The results of the DCCT are apparently applicable to NIDD with respect to microangiopathy, and hyperglycaemia is an independent risk factor for cardiovascular disease in NIDD patients. Strict glycaemic control is often associated with improvement in certain risk factors (lipids, hemorheology). Despite the fact that no large controlled prospective study similar to the DCCT is currently available for NIDD, efficient insulin therapy ensuring good glycaemic balance should be performed in these patients. However, the difficulties inherent to the implementation of intensive insulin therapy during NIDD should not be neglected: hypoglycaemic risk, weight gain, problems in elderly subjects, difficulties in instructing patients, and follow up. Finally, the return to adequate glycaemic control should be coordinated with an overall care plan for risk factors relative to macroangiopathy. PMID- 9410552 TI - [From the concept of fast acting analogs to the properties of the insulin Lispro]. AB - The difficulties of achieving good glycaemic control in insulin-dependent diabetes are due in large part to the inadequacies of subcutaneously administered insulin. In particular, resorption with the long acting form is variable from one subject to another and from one day to another and irregular over time, whereas the action of the rapid acting form is too late and prolonged. The slowness of absorption of the rapid acting form is attributable to the need for hexamer dissociation. The Lilly Laboratories, by inverting the amino acids lysine and proline in positions 28 and 29 in the B chain, have created an insulin (Lispro) which more rapidly dissociates into monomers after injection. The stability of Lispro is good, probably because of its phosphate buffer. In our experience, in conditions simulating use in portable insulin pumps, Lispro proved to be more stable than insulins specially intended for this use. The affinity in vitro was identical to that of insulin for its receptor. The affinity for insulin-like growth factor-I (IGF-I) receptor has been found to be 1.5 times as high as that of fast insulin in some models and comparable in others, and nearly 1,000 times less than that of IGF-1. Studies on in vivo potency and ex vivo cell growth, as well as of tolerance in the animal (mutagenicity, toxicity and carcinogenicity), have not shown a different effect from regular insulin (contrary to results for analogue Asp B10). The pharmacokinetics has shown an earlier and higher insulinaemic peak and a more rapid return to baseline values than regular insulin. On the basis of pharmacokinetic studies in normal subjects and diabetic patients, the characteristics retained by the licensing authorities are onset of action at 15 min, insulinemic peak between 30 and 70 min, and duration of action 2 to 5 h. Some clinical studies have shown a shortened action period of 1 to 3 h as compared to regular insulin and less influence of dose and injection site, notably with a return to normal insulin levels. The time required for normalisation is increased by 1 h if the injection is made in the thigh rather than the abdomen, as compared to 2 to 3 h for conventional insulin. This suggests that Lispro should be administered just before the meal (0 to 15 min). In some patients, an insulin with prolonged action can be added if the interval between injections is prolonged, i.e. always at the evening meal but possibly also at the noon meal. Lispro can be mixed with Umuline NPH or Umuline zinc without any alteration in its pharmacokinetics and potency if the injection is performed immediately. The few studies that have considered glycaemic stability and reproducibility have shown a tendency toward improvement in glycaemic excursions during the day, as measured by MAGE, and in insulinaemia variability expressed in area under the curve, which was reduced by half in the same individual or from one individual to another, with less marked impact on the variability from one day to another of glycaemic excursions. On the whole, Lispro provides faster kinetics, greater stability and possibly better reproducibility than fast insulin. These advantages, if confirmed by clinical experience, should allow an improvement in the comfort and glycaemic stability of diabetic patients. PMID- 9410553 TI - [The insulin analog, Humalog, in discontinuous: from pharmacology to clinical use]. AB - Humalog is the first analog of human insulin to be evaluated on a large scale in clinical conditions by discontinuous subcutaneous administration. Eight international multicentric studies have investigated the efficacy and tolerance of Humalog in protocols with 3 daily insulin injections (3 preprandial injections of Humalog associated with 1 or 2 injections of Umuline NPH or Umuline Zinc). A total of 2834 patients participated in these studies, including 2,277 who used Humalog. In addition to these trials, more than 5,000 patients had used Humalog by 1996, in some cases for more than 3 years. In insulin-dependent diabetes (IDDM), glycaemic control at the time of inclusion of patients in these protocols, as evaluated by HbA1C, was generally moderate (8 to 9%) despite 3 daily injections. This did not constitute intensive care since basal insulin for half of these patients was ensured by only a single daily injection. In these conditions, the results obtained showing significant postprandial improvement in glycaemic control and a significant reduction of the number of hypoglycaemic episodes, particularly at night, are important even though HbA1C levels were not significantly decreased. In a more recent crossover study (1996), 199 well controlled IDDM patients (HbA1C = 7.3%) were treated intensively by multiple injections. The number of severe hypoglycaemic episodes with coma was only one fifth that of the group treated by Humalog (equivalent to a reduction of 26 comas per 100 patients/year) in the absence of any significant modification of HbA1C. No differences were noted for lipids, adverse side effects or severe events (except hypoglycaemic episodes) and immunogenicity. In non-insulin-dependent diabetes, the results were similar but less impressive concerning the improvement in postprandial glycaemic control and the reduction in hypoglycaemic episodes. In addition to the significant results obtained in these studies, Humalog was favoured by the vast majority of patients. It is likely that Humalog will allow intensified treatment of diabetes in very favourable conditions, and that more patients will achieve the difficult goal of normalising glycaemia. However, a single injection of NPH or prolonged insulin is not sufficient for that purpose. Two daily injections will generally be necessary. PMID- 9410555 TI - [Nursing care of the addicted patient]. AB - As drug use in Canada rises and more and more intravenous drug users become HIV positive, nursing the addicted client takes on a whole new meaning. However, nurses may not have had the opportunity, during basic training, to cover the subject of substance abuse in depth. This article explores addictions nursing and provides information that will help nurses ensure quality nursing care for this population. The first part of the article provides statistics on drug consumption, followed by a classification of the various drugs and a definition of substance abuse. Nursing interventions to deal with clients' specific health needs are presented, as are innovative projects illustrating nursing achievements in the field. PMID- 9410554 TI - [Lispro analog and quality of life]. AB - Quality of life was assessed in a multicentre random cross-over study (UK Benelux) of 189 well-controlled IDDM patients undergoing treatment with lispro insulin analog (Lilly). Lispro offered several advantages over regular insulin for intensive diabetes treatment. Patients were more satisfied, felt more flexible in their lifestyle (no delayed meals due to injections, need for fewer snacks), reported a decrease in hypoglycaemic episodes, and considered their glycaemic control as better during the lispro period. Seventy-four percent of patients elected to continue treatment with lispro. A study of different injection times related to meals provided better knowledge of postprandial blood glucose excursions after lispro and showed that the optimal time for lispro injection was just before the meal, which was very convenient for the patients. Another study showed that lispro analogue, because of its pharmacokinetic properties, can reduce dietary restrictions in well-controlled IDDM patients on intensified insulin treatment. PMID- 9410556 TI - [Education in domestic violence]. AB - A baccalaureate-level, mandatory course on the social aspects of violence against women is being given as part of the curriculum at the Universite du Quebec a Rimouski. The course has been offered since 1990 and includes a complete array of teaching techniques such as lectures, simulations and role playing. Its aim is to impart knowledge, develop awareness, promote a change of attitudes and develop intervention abilities for nurses who may come in contact with domestic violence situations. Prior to implementation, an exploratory study was completed to determine student nurses' perceptions of domestic violence. Results indicate that, prior to taking the course, domestic violence was perceived as an individual problem. Respondents (26 female and 2 male) generally accepted society's prejudices of domestic violence as fact and ignored research results that pointed to the social realities. By the end of the course, the respondents' knowledge of the social aspects of domestic violence was better integrated. The authors conclude that specific training on domestic violence can modify false perceptions and help nurses develop the necessary competencies to deal with these situations. PMID- 9410557 TI - The road to Florence. PMID- 9410558 TI - A state of mind. AB - This essay traces the personal odyssey of a pediatric neurosurgeon from the time of the 'second generation' of subspecialty (the 1960s) to the present. The capture and appreciation of the ethos of the neurosurgical care of children is described and emphasized. It is not possible to overstate the importance of certain role models in this process, a process that does, indeed, induce a lasting 'state of mind.' Concerns for some current trends are raised. PMID- 9410559 TI - My memories of Japanese pediatric neurosurgery. AB - At the end of World War II, medical students in Japan knew that the United States of America were the world leaders in medical science, and they dreamed of establishing such a rational and pragmatic brand of medicine in their own country. As the number of motor vehicle accidents increased in parallel with the raid growth of the Japanese economy after the war, the need for neurosurgery also intensified. The number of papers dealing with pediatric issues increased tremendously after the ISPN and JSPN had first held meetings in Japan in September 1973. It seems mandatory for us to back up the activity of the children's hospitals by establishing a subspecialty of pediatric neurosurgery. PMID- 9410560 TI - One pediatric neurosurgeon's story. AB - The development of pediatric neurosurgery in the United Kingdom, as experienced by one recently trained neurosurgeon in the late 1940s and the 1950s, is described. PMID- 9410561 TI - An exclusive group of "pediatric neurosurgeons"?--No! AB - In the past several years there has been a movement created by a small group of neurosurgeons, who advocate the segregation of the practice of pediatric neurosurgery from the rest of neurosurgery, and the creation of separate training standards and even a qualifying board independent of the American Board of Neurological Surgery. Some thoughts on the matter are presented, and caution is advised. PMID- 9410562 TI - Proceedings of the National Conference on Emerging Foodborne Pathogens: Implications and Control. Alexandria, Virginia, March 24-26, 1997. PMID- 9410563 TI - [Compensatory enlargement and pathologic remodelling]. PMID- 9410564 TI - [Clinical role and implications of intracardiac atrial fibrillation]. PMID- 9410565 TI - [Is carotid stenosis still a plank for the surgeon in the percutaneous era?]. PMID- 9410566 TI - [Sodium-hydrogen exchange and reperfusion myocardial injury: possible pathogenetic role and therapeutic implications]. PMID- 9410567 TI - [Myosin isoforms in skeletal muscle in patients with chronic heart decompensation: distribution and correlation with with exercise tolerance]. AB - Chronic heart failure (CHF) is accompanied by a reduced exercise capacity, and the symptoms can be at least in part explained by qualitative and quantitative changes in the skeletal muscle composition and metabolism. We have correlated the myosin heavy chain (MHC) composition of the gastrocnemius in 20 patients with different degrees of CHF to expiratory gases measured during maximal cardiopulmonary exercise testing, NYHA functional class and echocardiographic parameters. MHC composition was determined electrophoretically in skeletal muscle needle microbiopsies and the percent distribution calculated by laser densitometry. There was no correlation between ejection fraction, left ventricular end-diastolic and end-systolic diameters and MHC composition. The percentage of MHC 1 (slow aerobic isoform) was positively correlated with peak VO2 (r2 = 0.5, p = 0.0004), ventilatory threshold (VT, r2 = 0.33, p = 0.008), and O2 pulse (peak VO2/HR, r2 = 0.40, p = 0.003). There was a negative correlation between MHC 2a and 2b (fast isoforms) and peak VO2 (r2 = 0.38 and 0.37, p = 0.004, respectively), VT (r2 = 0.2, p = 0.05; r2 = 0.34, p = 0.007, respectively) and O2 pulse (r2 = 0.39, p = 0.003; r2 = 0.23, p = 0.03, respectively). NYHA functional class was also negatively correlated with the same parameters (r2 = 0.2, p = 0.01; r2 = 0.4, p = 0.001; r2 = 0.34, p = 0.006, respectively) as well as with MHC 1 (r2 = 0.62, p = 0.0001). A positive correlation was found between NYHA functional class and MHC 2a and 2b (r2 = 0.46, p = 0.001; r2 = 0.41, p = 0.002, respectively). The severity of heart failure is paralleled by a shift of the MHC pattern toward the fast MHC 2b. The correlation between the magnitude of the MHCs shift, from the slow aerobic to the fast type, with both clinical parameters (NYHA functional class) and functional measurements (peak VO2, VT, O2 pulse) of exercise capacity seem to suggest that changes in skeletal muscle composition may play a key role in exercise tolerance in patients with CHF. PMID- 9410568 TI - [Value of peak oxygen consumption during exercise for the prognostic stratification of patients with severe systolic dysfunction of the left ventricle]. AB - We sought to assess the prognostic value of peak exercise oxygen consumption (peak VO2) in patients with severe left ventricular systolic dysfunction and mild to moderate symptoms of chronic heart failure. We focused on 1-year mortality. We prospectively studied 77 patients with left ventricular ejection fraction (EF) < or = 25% and NYHA functional class I/II (61%) or III (39%). All patients underwent cardiopulmonary exercise test, two-dimensional echocardiography and 24 hour Holter monitoring. Examined variables were age, etiology, NYHA functional class, EF, peak VO2, and presence of nonsustained ventricular tachycardia. Overall 1-year mortality rate was 23%. At univariate analysis, age > or = 60 years, ischemic etiology, and peak VO2 < 14 ml/kg/min were significantly associated with mortality. At multivariate analysis, peak VO2 was the most powerful predictor of death (p = 0.0001). In the subgroup of patients with a peak VO2 < 14 ml/kg/min, the actuarial 1-year mortality rate was 56%. One additional patient underwent heart transplantation because of severe hemodynamic deterioration. By contrast, in the subgroup of patient with a peak VO2 > 14 ml/kg/min, 1-year mortality rate was 11%. This study provides evidence that patients with severe left ventricular dysfunction and mild to moderate symptoms of chronic heart failure can be accurately stratified into subgroups with strikingly divergent prognosis by an objective criteria such as peak VO2. PMID- 9410569 TI - [Implantation of the Palmaz-Schatz stent in coronary vessels irrigating and non irrigating necrotic myocardial areas: comparison of acute results and at mid term]. AB - Recent evidence suggests that higher restenosis rate is observed after coronary angioplasty of an infarct-related artery. Furthermore, angiographic restenosis seems associated with a deterioration of left ventricular function at follow-up. The aim of this study was to assess the acute results and angiographic restenosis following coronary artery stenting of infarct-related (Group 1) and non infarct related coronary arteries (Group 2). We retrospectively analyzed the results of 381 consecutive patients treated with Palmaz-Schatz coronary stent implantation between May 1992 and January 1996. Stenting of the infarct-related artery was performed in 154 patients (Group 1), while 227 patients (Group 2) received stenting of the non infarct-related artery. Both groups had similar age, gender, clinical conditions and coronary angiographic pattern. There were no significant differences between groups, concerning type of stented coronary vessel (left anterior descending-LAD 52.4% vs non-LAD 47.6%, Group 1, LAD 59.5% vs non-LAD 40.5%, Group 2) and number of stents per patient (1.31 +/- 0.48 in Group 1, 1.18 +/- 0.56 in Group 2) and per coronary vessel (1.17 +/- 0.54 in Group 1, 1.09 +/- 0.46 in Group 2). The procedure was performed using similar maximal inflation pressures in both groups (13.3 +/- 2.9 atm in Group 1, 13.40 +/- 3.17 atm in Group 2). Technical success was achieved in 96.8% of Group 1 and in 96% of Group 2 patients. Acute coronary stenting success and major adverse events (acute myocardial infarction, emergency bypass, death) were similar in both groups of patients. No difference was found in restenosis rate at 6-month angiographic follow-up (Group 1 = 29.8%, Group 2 = 27%). In conclusion, this study indicates that stenting of infarct and non infarct-related coronary arteries has similar success and 6-month restenosis rates. PMID- 9410570 TI - [Univariate analysis of potential risk factors for early mortality (within 28 days) after aortocoronary bypass in Italy. OP-RISK Study Group]. AB - The multicenter OP-RISK study, developed during 1994-96, was aimed at: 1) investigating early (28 days) death rates following aortocoronary bypass surgery among patients recruited from four Centers representing geographical distribution in Italy; 2) defining possible risk factors for early mortality, also comparing these factors with those reported in previous studies. Average values are reported and compared of 65 variables (36 preoperative, 10 operative and 19 postoperative) out of 984 patients subdivided into alive (n = 940) or dead (n = 44, 4.47%) at 28 days (155 +/- 174 hours, interval between 12 and 576 hours) postoperatively. Causes of death were cardiac in 37 (77%), pulmonary in 3 (0.7%), vascular in 2 (0.5%) and infective in 2(0.5%) patients, respectively. During the study a total of 1126 patients were operated upon in the collaborative Centers with the diagnosis of coronary artery disease and 51 deaths were reported officially in-hospital (4.53%). Therefore, OP-RISK data represent 87% of overall patients and a superposable death rate. The potential role as risk factors of early mortality was assessed univariately for 17 preoperative, 5 operative (in 3 cases for the first time) and 5 postoperative factors. In general, it was confirmed that factors defining left ventricular function are sensitive predictors of mortality. In OP-RISK we were able to show, in addition, that tachycardia (> 130 b/min) at induction of anesthesia, and total time of anesthesia, cardiopulmonary bypass and aortic cross clamping may be significant factors among operative variables as might be among postoperative ones several arrhythmia types or a lower rate in antithrombotic therapy with aspirin at 6-12 hours postoperatively. The protective role of bypass surgery performed with at least 1 arterial segment was also ascertained. Most of these potential factors were significantly related to outcome (either directly or inversely) as were among them, as seen in a subsample (65%) of 639 patients in whom a correlation matrix was performed among 16 factors selected on the basis of the common denominator principle. Our results suggest that it is possible to collect in a multicenter experience univariate predictors of early mortality following aortocoronary bypass surgery in Italy, which are not different from those reported from previous studies performed abroad. Operative indicators may also have predictive capabilities. The effort may be worthwhile and demands further cooperative studies to be undertaken, aimed at obtaining nationwide coefficients of risk along with representative average values of factors that soon might emerge once multivariate statistics will be performed on this material. PMID- 9410572 TI - [The pulmonologist]. PMID- 9410571 TI - [Anatomo-functional effects of ionic dilution in the isolated and working heart in the rat]. PMID- 9410574 TI - 23rd Annual meeting of the International Neuropsychological Society. Seattle, Washington, February 8-11, 1995. Abstracts. PMID- 9410575 TI - 17th Annual International Neuropsychological Society European Conference. Angers, France, June 22-25, 1994. Abstracts. PMID- 9410573 TI - G proteins and second messengers in psychiatry. PMID- 9410576 TI - NeuroNames Brain Hierarchy. AB - The NeuroNames Brain Hierarchy is a structured system of neuroanatomical terminology that provides a comprehensive representation of virtually all human and nonhuman primate brain structures that are identifiable either grossly or in Niss1-stained histological sections. This system was devised for computer applications to address deficiencies in the brain terminology presented in Nomina Anatomica. English terms are listed for 783 structures in nine levels of hierarchical ranking. Abbreviations are provided for all superficial and primary volumetric structures. The substructures that constitute the total volume of every superstructure are identified. Superficial features of the brain are clearly distinguished from internal, volumetric brain structures. Structures found solely in either humans or macaques are identified. The purpose of the NeuroNames Brain Hierarchy is to bring greater standardization to the neuroanatomical terminology used by scientific investigators, clinicians, and students. This effort is consistent with the goals of the Unified Medical Language System program of the National Library of Medicine. It is hoped that the systematic construction of the NeuroNames Brain Hierarchy will facilitate use of the most widely accepted definitions of classical neuroanatomy in quantitative computerized neuroimaging applications. It should provide an accurate structural framework against which to reference the many other kinds of neuroanatomical information that are acquired by modern imaging, mapping, and histological labeling techniques. PMID- 9410577 TI - Neuroinformatics. Proceedings of a workshop. Arlington, Virginia,USA, September 19-20, 1995. PMID- 9410578 TI - Functional MRI and brain disorders symposium. Bethesda, Maryland, November 15, 1996. PMID- 9410579 TI - Emerging Treatments in Multiple Sclerosis. Proceedings of the European Charcot Foundation Symposium. Brussels, Belgium, 23-24 November 1995. PMID- 9410580 TI - Models of Care in Multiple Sclerosis. Atlanta, Georgia, USA, September 27-29, 1996. Abstracts. PMID- 9410581 TI - Pathobiology and Treatment of Multiple Sclerosis. Proceedings of a symposium in honor of Dr Donald Silberberg. Pennsylvania, 20 October 1995. PMID- 9410582 TI - Retraction of paper: Two visual pigment opsins, one expressed in the dorsal region and another in the dorsal and the ventral regions, of the compound eye of a dragonfly, Sympetrum frequens (Invertebrate Neuroscience,1,33-39,1995) PMID- 9410583 TI - NEISS and child injury data. PMID- 9410585 TI - The 1996 Canadian outlook on gastroesophageal reflux disease. Consensus Conference. Ottawa, Ontario, June 1996. PMID- 9410584 TI - Recent advances in phage display: the report of the Phage Club first meeting. PMID- 9410586 TI - [Multifactorial analysis of risk factors for low birth weight in Salvador, Bahia]. AB - This study is a multifactorial analysis of the risk factors for low birthweight in a group of newborns in an urban area of Brazil. A total of 1023 infants born in four maternity units in Salvador, Bahia, between July 1987 and February 1988 were included in the study. The sources of information were clinical histories and interviews with the mothers in the maternity units. The analysis was by means of logistic regression. In the final model the risk factors were the following: maternal age less than 21 years or more than 35; gestational age less than 38 weeks; unfavorable outcome of an earlier pregnancy; interval of 12 months or less since prior birth; tobacco smoking; and hypertension. The population attributable risk values for the risk factors included in the final model are presented. These factors should be used to identify pregnant women at high risk of giving birth to a low-birthweight baby, in order to provide them with more prenatal care. PMID- 9410587 TI - [Communication and interactive education in health and its application to the management of the diabetic patient]. AB - This article analyzes the need for participation of the groups that are the recipients of health messages in order to make health education more effective. Taking the traditional educational approach as the point of departure, the different options for communication and education processes in this field are explored in depth. The participatory communication model, in which the subject plays the role of both transmitter and recipient, leads to the coproduction of health education messages. In light of the experience of the Diabetes Education Program of the National Endocrinology Institute of Cuba, a health education model based on a strategy of interactive communication is presented. Its objective is to develop in the patient the capability to think and decide about his or her health through discussion groups facilitated by a health care provider. In this model, the beneficiaries of the health message help shape its aim and content. The discussion centers on the patient, taking into account his or her real health needs and feelings. The results obtained with this methodology are reviewed, and it is concluded that this model opens new educational possibilities that will facilitate decision-making with regard to health and healthy lifestyles. PMID- 9410588 TI - [Distribution of human resources in 8 general hospitals in Sao Paulo]. AB - Technological development and the appearance of new medical specialties have led to increased division of labor in hospitals, with the resultant creation of subareas. Therefore, the redistribution of human resources in hospitals has become an important aim in order to achieve efficiency. On the basis of interviews conducted between May and October 1993 with the directors and superintendents of eight hospitals in Sao Paulo, Brazil, this study describes and analyzes the distribution of human resources in four broad areas: infrastructure, clinico-surgical inpatient care, the complementary area of diagnostics and therapeutics, and emergency and outpatient care. The study also analyzed the ratio of employees per bed, besides outlining a comparison of public and private hospitals. The results revealed that, on average, the ratio of employees to beds was 6.8. The proportional distribution of employees by broad areas was found to be 46.9% in infrastructure, 10.7% in diagnostics and therapeutics, 32.0% in inpatient care, and 10.3% in outpatient/emergency care. PMID- 9410590 TI - [New challenges in the field of advanced education for nursing personnel in Latin America]. AB - This document examines and describes the basic characteristics of the 48 postgraduate nursing programs that exist in Latin American countries. A critical and holistic analysis model is used to identify their ideological foundations, the policies that govern them at the ministerial and institutional levels, and the conditions in which the programs operate, in an attempt to highlight areas where critical needs exist and to offer recommendations on how to satisfy them. Emphasis is placed on the capacity of international agencies, governments, and universities to improve postgraduate nursing education through the definition of priorities in light of the general reforms taking place in the health sector, the provision of support for research on nursing topics, and the coordination of cooperation initiatives at the national and international levels. PMID- 9410589 TI - [Turnover of nursing staff in hospitals of the municipality of Ribeirao Preto]. AB - The present study was carried out in 1990 with the objective of measuring and analyzing the turnover of nursing personnel in nine general hospitals located in Ribeirao Preto, Sao Paulo State, Brazil. The population under study consisted of nurses, nursing assistants, nursing technicians, and nursing attendants who started work or left their jobs between 1 January and 31 December 1990. Employment status for these workers was checked monthly based on information from the human resources department of each hospital. Two types of turnover indices were calculated: the liquid rate of substitution and the median length of service of the departing workers. Results showed that the number of nursing personnel employed in Ribeirao Preto increased in 1990. However, results also revealed a high turnover rate, especially for philanthropic and private institutions, with liquid substitution rates of 32% and 39% per year and median lengths of service of 8 months and 12 months, respectively. These circumstances could lead to higher costs and reduced productivity. The only public hospital in the study presented the lowest level of turnover (liquid substitution rate of 6% and median length of service of 42 months). Nurses and attendants had the highest substitution rates and thus were the least stable categories. PMID- 9410591 TI - Building confidence by profiling. PMID- 9410592 TI - [Encouraging organ donation: the paradox]. PMID- 9410593 TI - Violence and the nursing curriculum: nurse educators speak out. AB - Violent crime in the United States is increasing at an alarming rate. Nurses are the first to encounter its victims--but are they prepared for them? Two activities at the 1995 NLN convention gathered the opinions, concerns, and recommendations of nurse educators about how education can help better equip nurses to perform the role of violence preventer and care provider. PMID- 9410594 TI - [Peripheral autonomic disorders in patients with the initial manifestations of cerebral blood supply insufficiency]. AB - 267 patients with vascular encephalopathy and initial manifestations of cerebrovascular insufficiency as the result of hypertension and cerebral atherosclerosis were observed by means of complex vegetological methods. The signs of peripheral autonomic insufficiency were revealed in 89% patients with vascular encephalopathy and in 64% ones with initial manifestations of cerebrovascular insufficiency. Cardiovascular tests directed to evaluation of the autonomic insufficiency were suggested to use for the prognosis of decompensation of cerebrovascular blood supply in such patients. PMID- 9410595 TI - [The cerebellar cortical atrophy syndrome]. AB - The results of the observation of 44 patients with cerebellar syndrome of different etiology are presented. Together with careful study of anamnestic and clinical data some additional examinations were performed: senso- and pallesthesiometry, thermovisional investigation, vestibulometry, electroencephalo , electromyography, computer tomography. The study allowed to reveal both the cause of the disease and to refer etiologically late cerebellar cortical ataxia to alcohol factor. On the basis of the comparison of clinical neurological data with paraclinical observation differential diagnostic criteria were defined for Marie-Foix-Alajouanine's late cortical cerebellar atrophy in alcoholism, in the cases of the hereditary predisposition as well as of unclear genesis, in Holmes olivocerebellar atrophy, in Menzel, Hunt and Dejerine-Thomas olivopontocerebellar degeneration. PMID- 9410596 TI - [The forms of alcohol abuse and disorders of cardiovascular activity in the dynamics of the alcohol abstinence syndrome]. AB - 55 individuals were observed (41 patients with alcoholism and 14 healthy controls). The patients were divided into 2 groups: patients with permanent alcohol abuse (the 1-st), and patients with periodic alcohol taking (the 2-nd). Heart rate, systolic and diastolic arterial pressure were measured. Echocardiography was performed too. In the 1-st group disorders of myocardial relaxation were revealed in the structure of withdrawal syndrome with prevalence of blood flow in the phase of early left ventricular filling and moderate increase of contractility of the left ventricle. Asymptomatic course of alcohol withdrawal syndrome was quite characteristic for the first group too. In the 2-nd group (drinking, pseudodrinking alcoholism) there were disorders of myocardial relaxation with prevalence of blood flow in the late phase of left ventricular filling and moderate elevation of general contractility of the left ventricle. There were also complaints concerning disorders of cardiovascular circulation. It was revealed that alterations of cardiovascular activity in periodic form of alcohol administration manifested more pronouncedly during the first days of alcoholic withdrawal syndrome and disappeared more quickly as compared with permanent drinking. The usage of 3 groups of drugs in management of alcoholic withdrawal syndrome was grounded depending on the form of alcohol administration: drugs which increase the activity of cerebral inhibitory transmitters' systems; drugs which prevent increased activity of sympathoadrenal system; drugs which influence energetic and chemical homeostasis. PMID- 9410597 TI - [Autonomic nervous system function at different periods in the alcohol abstinence syndrome]. AB - Four periods of alcoholic withdrawal syndrome development were identified during examination of 156 male patients: 1-5 days (the 1st period), 6-10 days (the 2nd), 11-30 days (the 3rd), 30 days and more (the 4th). Different indices of autonomic nervous system function were investigated including different tests (cold test, ocular-cardiac reflex, ortho- and clinostatic tests, psychological tests, etc). It was determined that sympathicotonia which was quite characteristic for the 1st period became less expressed in the 2nd one. The 3rd period was characterized by maximal parasympathetic prevalence, especially in cardiovascular system. The 4th period was characterized by normalization of the basic autonomic indices with partial vagotonic tendencies. PMID- 9410598 TI - [Symptomatic alcoholism and remissions in schizophrenia in light of a 20-year catamnesis]. AB - 80 patients aged 14-60 years with progredient paranoid schizophrenia which began alcohol abuse after the first attack of the disease were observed. The peculiarities of both clinical manifestations and course of alcoholism and schizophrenia were analysed in age aspect. The patients were divided into 3 groups according to age: 14-20, 21-40 and 41-60 years. It was established that alcohol abuse in schizophrenia does not cause any further increase of alcoholic symptomatology and had not the phased development as in alcoholism, i.e. alcohol abuse does not cause typical alcoholic disease. Alcoholism, which is symptomatic in schizophrenic patients, can change, however, the clinical picture of psychosis. It can cause atypical and soft course of the disease or, on the contrary, development of decompensation in remission and increase of psychic defect. PMID- 9410599 TI - [Mental disorders in the offspring of alcoholic fathers (children from an early age to 28)]. AB - The examined groups included 175 children whose fathers were alcoholics and 73 individuals from nonalcoholic parents. The study group was divided by age into children from infantile age to 15 years and 16-28-year-olds. The wide range of psychopathologic disorders were revealed in children up to 15 years of age: syndrome of hyperactivity, neurotic disturbances, mental retardation, epileptic like syndrome. Alcoholism was found in sons only (3.77%). Mental disorders were revealed significantly more frequently in sons of alcoholic fathers than in their daughters. Their incidence increased with the age of the sons. The disturbances occurred significantly more frequently in the test group as compared with the control one (19 and 6.38%, respective by). In the older group (16-28-year-olds) the spectrum of mental disorders consisted of alcoholism, mental retardation, psychopathic-like and neurotic disorders. Alcoholism occurred most frequent and its rate depended on the age and sex. Frequency of alcoholism was 66.7% in sons aged 21-28 years and 15.38% in daughters of the same age. The conclusion was made about multiple risk for children of alcoholic fathers. More vulnerable contingent was the sons for which the risk of alcoholism was very high. PMID- 9410601 TI - [Magnetic and electrical stimulation in the rehabilitative treatment of patients with organic nervous system lesions]. AB - 89 patients with organic damages of nervous system with paralyses and pareses as the main symptoms in clinical pattern were treated. Their treatment was complex with application of impulse magnetic field and electrostimulation which permitted to achieve multilevel electrostimulation. The control group was formed by 49 patients with analogous diseases which were treated by sinusoidal current electrostimulation only. Combined application of magnetic stimulation and electrostimulation was more effective. That was confirmed by data of roentgenographic and electromyographic studies. PMID- 9410600 TI - [The current status of the drug abuse problem in Russia]. AB - The statistic data evidence for rapid increase of the number of narcomanias, especially in juveniles and young people nowadays in Russia. The spectrum of psychoactive preparations used became also wider. On the basis of medico biological studies the conception concerning main pathogenetic mechanisms of the development of psychoactive substances dependence was formulated. Enzyme immuno assays either for diagnosis of long-term opiate administration or for determination of blood serum methadone level were elaborated. Clinical studies demonstrated pathomorphism of the symptoms of dependence upon well-known narcotics. Clinical pattern of dependence upon new psychoactive preparations which cause dependence syndrome was also investigated. New differentiated methods of treatment of various types of narcomanias are proposed. There is underlined that a comprehensive program of narcomanias prophylaxis in Russia is needed. PMID- 9410602 TI - [Hemocarboperfusion in the treatment of patients with an opiate abstinence syndrome]. AB - The authors analyze of the results of applying hemocarboperfusion in treatment of opiate addiction. All the patients were treated in the Belorussian center of sorption methods of detoxication and plasmapheresis. It is shown that 4-5 procedures of hemocarboperfusion in combination with infusion and drug therapy allow to correct of withdrawal mental and somatic disorders. PMID- 9410604 TI - [Aphasia]. PMID- 9410605 TI - [Myorelaxants in the rehabilitation of patients with poststroke motor disorders]. PMID- 9410603 TI - [Clinico-biochemical study in epilepsy]. AB - Diagnostic opportunities of the test of paroxysmal activity (TPA) were examined in 253 epileptic patients of different age and duration of disease. It was established that the changes of autoantibodies (aAB) titers in the control group were about 64-110% (mean value 85%). Meanwhile in epileptic patients there was elevation of the same indices 2.1 times (182%). The correlation was observed between the levels of aAB titers and duration of the disease, frequency and character of paroxysmal disorders. The conclusion was made that TPA may be used quite successfully in diagnosis of epilepsy, especially in difficult cases. The level of accumulation of aABs in blood of patients with the fits may be used as new immunological index which may facilitate essentially objective of diagnosis of epilepsy. PMID- 9410606 TI - [The efficacy of Solpadeine in patients with primary forms of headache]. PMID- 9410607 TI - [The use of antioxidants in drug abuse (preliminary data)]. PMID- 9410608 TI - [The EEG spectrogram of alcoholic patients]. PMID- 9410609 TI - [Pentoxifylline in the treatment of disorders of the cerebral circulation]. PMID- 9410610 TI - [Cardiac involvement in alcoholic patients]. PMID- 9410611 TI - [Immunohistochemical expression of p53 protein in squamous epithelial carcinomas and normal and dysplastic epithelium of the mouth cavity]. AB - Routinely formalin-fixed and paraffin-embedded material of 123 squamous cell carcinomas of the floor of the mouth and tongue bordered by non-tumorous mucosa were immunohistochemically investigated for the expression of p53 protein using a panel of four anti-p53 antibodies (CM1, PAb 1801, DO7, PAb240) following wet autoclave antigen retrieval. p53 immunoreactivity was detected in 55-67% of the carcinomas, with the different antibodies used showing a preferential accumulation of the p53-positive tumour cells at the invasive tumour front. In dysplastic and normal epithelium focal or dispersed p53 immunopositivity could be detected in 62-72% and 57-70% of the cases, respectively, regardless of the p53 immunostatus of the carcinomas. No statistically significant correlations between p53 immunophenotype of the tumours and clinico-pathological parameters, or survival of the patients, could be detected statistically. It is concluded that immunohistochemically detectable p53 accumulation in oral cancer might indicate an early stage of carcinogenesis. The immunohistochemical detection of p53 in these tumours, however, is without prognostic significance. PMID- 9410612 TI - [Cytokines and cytokine receptors in mouth mucosa of immune suppressed patients]. AB - In vitro studies on epithelial cells suggest that cyclosporin (CsA) inhibits a pathway of production of IL-6, which is mediated by TNF alpha and E. coli. The aim of the present immunohistochemical study was therefore to evaluate the content and location of cytokines and cytokine receptors in the oral mucosa of 10 patients with CsA medication. Ten patients without immunosuppression served as controls. Specimens were procured during surgical treatment and processed by paraffin embedding and as cryosections. Expression of TNF alpha, IL-1 beta and IL 6-receptor was present both in the epithelial layer and the submucosal tissues, but did not differ very greatly between patients with CsA therapy and the controls. IL-6, however, displayed a positive reaction only in the control group, while it was invisible in the group of patients with CsA medication. The immunohistochemical findings of the present study suggest that there may be a lack of mucosal expression of IL-6 under CsA medication. With regard to the broad spectrum of possible effects of proinflammatory cytokines and their multiple interactions, the functional relevance of these findings merits further investigation. PMID- 9410613 TI - [Osteomyelitis of the facial skull in Albers-Schonberg osteopetrosis]. AB - Albers-Schonberg osteopetrosis, a rare heritable bone disease with autosomal dominant or recessive transmission, is generally characterised by diffuse sclerosis of the whole skeleton accompanied by pathological bone fragility and delayed physical development, profound intractable myelophthisic anaemia, neurological deficits, and osteomyelitis, especially of the jaws and the skull. The precise aetiology of the osteopetrosis is not clear. Therefore therapy is restricted to alleviation of symptoms. In this study the case of a patient suffering from the benign form of osteopetrosis is presented. Osteomyelitis of the skull was treated successfully 2 years after the removal of lower and upper jaw teeth by a combined multistage surgical and antibiotic approach. PMID- 9410614 TI - [Report on the 27th Annual Meeting of the German-Austrian-Swiss Working Group for Tumors of the Maxillofacial Area]. PMID- 9410615 TI - [Oral manifestations of autoimmune diseases with sicca symptoms. Comparative studies of patients with primary and secondary Sjogren syndrome]. AB - Sjogren's syndrome is a chronic autoimmune disease which mainly affects exocrine glands. We present a study where 123 patients with Sjogren's syndrome were examined according to their oral symptoms. A high percentage of patients showed mild or moderate inflammation of the oral mucosa. Remarkable is the high number of extracted teeth in older patients with Sjogren's syndrome. Oral ulcerations and erythema were found only in the group with secondary Sjogren's syndrome and systemic lupus erythematosus. PMID- 9410616 TI - [HIV-associated salivary gland diseases. Review of the literature and 3 case reports]. AB - HIV-associated salivary gland disease (HIV-SGD) includes lymphoepithelial lesions and cysts involving the salivary gland tissue and/or intraglandular lymph nodes, and Sjogren-like conditions. Three cases of salivary gland disease occurring in HIV-infected patients are reported. Histopathological examination showed squamous epithelium-lined cysts. In the walls of the cysts lymphoid tissue, epitheloid granulomas and giant cells were found. The clinical and histopathological criteria as well as magnetic resonance imaging and therapy are discussed. PMID- 9410617 TI - [Focal oncocytosis of the salivary glands and etiopathogenetic relations]. AB - Focal oncocytosis in minor and major salivary glands is a relatively rare condition. We found it in 92 (8.7%) of 1056 biopsy and operation specimens of the salivary glands. There was a nearly balanced sex ratio. Oncocytosis was found mainly in patients between the 5th and 8th decades of life, with a progressive tendency. Oncocytic transformations of the ductal epithelium were always associated with additional lesions. These were chronic sialadenitis, hyperplasia and metaplasia of ductal epithelium and morphological signs of central or peripheral salivary obstruction. As etiologic factors or fundamental diseases, mainly neoplasms and chronic inflammations with hyperplasia of the oral mucosa and a primary chronic (obstructive) sialadenitis were found, but age-related mechanisms could not be excluded. We believe the oncocytic transformation to be a regressive alteration of previously hypertrophic or hyperplastic ductal epithelium with the appearance of a mitochondriopathy. PMID- 9410618 TI - [Oral manifestations in patients with systemic lupus erythematosus]. AB - Forty-six patients with systemic lupus erythematosus underwent thorough dental examination to determine the frequency and severity of oral lesions and periodontal diseases. According to clinical criteria, disease was classified as severe (n = 26) or less severe (n = 20). The overall rate of mucosal involvement in the studied patients was 48%-from 54% in patients with severe disease, 40% in those with less severe disease. Patients with severe disease were found to have a higher rate of tooth loss and an increased rate of gingival inflammation. The severity of periodontal lesions correlated with alterations in the immunoglobulin pattern, particularly with an increase in gamma-immunoglobulins. Thus it is suspected that complex immunodysregulation in combination with immunosuppressive therapy is responsible for the high rate of oral and periodontal lesions in patients with systemic lupus erythematosus. PMID- 9410619 TI - [Immunocytochemical venous blood studies in patients with manifest oral cavity carcinomas, oral precancerous conditions, benign tumors and in chronic alcoholic patients]. AB - In a prospective pilot study we investigated the percentage of immunocompetent cells in the peripheral blood in 146 patients (lymphocytes, leucocytes, monocytes, T cells, B cells, NK cells, T-helper cells, T-suppressor cells, ratio T-helper/T-suppressor cells, activated T cells HLA-DR) by flow cytometry. The immunologic parameters were derived from patients with oral and oropharyngeal squamous cell carcinomas, precancerous lesions and benign tumours and from a group of heavy smokers and alcoholics. Carcinoma patients (n = 46) were compared with risk groups and a reference group consisting of patients with inflammatory disease. Within the collective of carcinoma patients we measured the immune status before and after chemo-, radio- and operative therapy. We also analysed the immune parameters in relation to clinical and histomorphological parameters (TNM status, grading). The univariate analysis of monocytes showed significant relationships between on the one hand carcinoma patients and on the other alcoholics and those with benign tumours and precancerous lesions. In precancerous lesions NK cells were significantly increased compared with alcoholics and the reference group. A significant decrease in B cells in carcinoma patients may show incipient insufficiency of the humoral immunity. The immune parameters showed a different reaction depending on therapy. After irradiation we found a significant increase of T-suppressor cytotoxic cells and decreases in B and T-helper cells. Chemotherapy showed an increase in T and T helper cells and a decrease in B cells. Surgical therapy alone yielded an increase in B cells. The comparison of all pre- and posttherapeutic parameters showed significant changes only in activated T cells HLA-DR. We found no correlation between prognostic clinico-pathological factors and immune parameters. No changes were found in a multivariate analysis. PMID- 9410620 TI - [Expression of proliferation markers PCNA and MIB 1 in verrucous squamous epithelial carcinoma of the oral cavity]. AB - Expression of proliferation markers PCNA and MIB 1 was evaluated in 17 verrucous carcinomas and in five cases of normal squamous epithelium of the oral cavity. Immunohistochemistry revealed no significant differences in the proliferative activities. Highest expression was seen in the basal zones. Proliferative activity in more than 26% of cells is no longer compatible with verrucous carcinoma, but favours the diagnosis of squamous cell carcinoma. PMID- 9410623 TI - [Manifestation of a highly malignant B-cell lymphoma simulating osteomyelitis of the mandible]. AB - A case of precursor B-cell lymphoma of unusual location in the mandible is presented. Clinical features as well as technical examinations led to the misdiagnosis of chronic osteomyelitis. Only immunohistological examination of intraoperatively taken biopsies was able to reveal the true diagnosis. High-dose chemotherapy was started and full remission could be achieved. PMID- 9410621 TI - [Basaloid squamous epithelial carcinoma of the mouth mucosa]. AB - The basaloid-squamous carcinoma (BSC) that was first described in 1986 by Wain et al. for the head and neck region is a rare distinct variant of squamous cell carcinoma (SCC). The cardinal histopathologic feature is a biphasic cellular pattern of basaloid and squamous components. BSC has been confused with solid adenoid cystic carcinoma (ACC). Although the number of reported cases is small, BSC appears biologically virulent, with a propensity to aggressive local behavior, early regional and distant metastasis, and subsequent poor survival. We report the clinicopathological characteristics of 4 new cases and compare their immunohistochemical features with those of solid ACC and conventional SCC. Our results show that BSC, ACC and SCC react to CK 5/6. SCC is CK 10- and CK 13 positive, while BSC and ACC are negative for these markers. BSC and ACC react to CK 8, but in ACC only the luminal cells are CK 8 positive: therefore ACC has a glandular pattern. Our findings indicate that the immunohistochemical differences between BSC and ACC can facilitate their differential diagnosis. Because the biologic behavior of BSC differs from ACC and SCC, distinction among these tumor types is warranted. PMID- 9410622 TI - [Detection of human papillomavirus (HVP)-DNA in oral manifestation of lichen planus]. AB - Human papilloma viruses (HPV) can be detected in different epithelia with the help of the polymerase chain reaction (PCR). The role of HPV in the development of anogenital cancers has been intensively studied, and current evidence shows that most cervical cancers are associated with so-called high risk HPV types (e.g. HPV 16 and 18). HPV-infections can also be demonstrated in oral premalignant lesions and squamous cell carcinomas. Depending on the sensitivity of the detection method, 40-67% of leukoplakias, 2.5-76% of squamous cell carcinomas and 0-87% of cases of lichen planus were described to be infected with HPV 16 or 18. Whether lichen planus can be considered as a premalignant lesion is still controversial. By the use of PCR and hybridization we found infections with the high risk HPV types 16, 18 and 31 in 42% (3/7) of the patients with lichen planus. Further investigations with a higher numbers of cases in combination with the analysis of the viral gene expression as well as the clinical and histological control of the corresponding regions are necessary. The aim of these studies is to find out the prognostic value of the HPV infection for this facultative premalignant disease. PMID- 9410624 TI - [Acoustic reflection position monitoring of tracheal cannulae. A contribution to quality assurance?]. AB - Non-invasive acoustic airway monitoring was evaluated in an experimental study. Recording amplitude and travel time of acoustic pulse response, an area-distance function of the cross sectional dimensions of the endotracheal tube and the adjacent airway was calculated to obtain an acoustic pattern of the airway. Measurements on models and excised human cadaver lungs were performed to discover whether displacement or obstruction of the artificial airway can be detected in the acoustic equivalent. Regression analysis revealed a close correlation between displacement of tracheostomy tubes and the shifting of the acoustic area-distance function (corr. coeff. 0.97-1). Dispersion analysis confirmed reasonable reliability (coeff. of variation 0.6-2.1%). Location and amount of obstruction could likewise be identified. Thus acoustic mapping provides an adequate approximation of the true geometry of tracheostomy and endotracheal tubes. We conclude that acoustic monitoring may provide a powerful tool to achieve primary prevention of airway disturbances in intubated and mechanically ventilated patients, as geometrical changes of airway configuration can be detected even before they cause substantial effects on respiratory metabolism. PMID- 9410625 TI - [Median facial dysplasia in patients with lip-jaw-palate clefts. Characteristics and problems in interdisciplinary orthodontic-oral surgery treatment]. AB - The diagnosis of median facial dysplasia is made in cases of a typical deficiency of the median facial structures without concomitant abnormalities of the brain, and presence of unilateral or bilateral cleft lip and palate. The identification and description of this type of cleft lip and palate are important because such patients have severe growth disturbances and tissue deficiencies. These make surgical reconstruction and orthodontic treatment difficult. This is demonstrated in two examples. The clinical and roentgenological findings in the mother of one of the demonstrated patients point to a discrete form of the same anomaly. PMID- 9410626 TI - [Lateral orbitectomy for correction of endocrine ophthalmopathy]. AB - Grave's disease is often associated with various symptoms summarized as endocrine orbitopathy. A surgical procedure is presented which reduces the exophthalmos by approximately 4.5 mm by resecting the caudal and lateral orbital wall through a subciliar incision. The surgical intervention regularly leads to an amelioration of the functional symptoms of endocrine orbitopathy, such as diplopia, corneal exposure and lid irregularities. In one case an acute reduction of vision on both eyes significantly ameliorated immediately after surgery. The operation described also improves the aesthetic appearance of the patients, who often suffer psychologic impairments associated with Grave's disease. PMID- 9410627 TI - [Ameloblastic fibroma of the maxilla]. AB - Ameloblastic fibromas are rare benign odontogenic tumours, which appear preferentially in adolescents and young adults. They are most often found in the mandible. The present case report describes a 40-year-old patient with an ameloblastic fibroma in the upper jaw, an extremely rare site. The diagnosis, differential diagnosis, histology and therapeutic procedure are described. PMID- 9410628 TI - [Aggressive growth of hemangiopericytoma in the area of the facial skull]. AB - Hemangiopericytoma is a rare tumor that originates from the pericytes. Only histology permits a reliable diagnosis. There are no known parameters to predict the biological behavior of the tumor, so every hemangiopericytoma has to be treated as potentially malignant. No age or gender prevalence of this tumor in the region of the head neck has yet been observed. Hemangiopericytoma should be treated by radical surgery; chemotherapy or radiation should be reserved for incompletely removed or metastatic tumors. Metastasis and recurrences have been described even decades after first tumor treatment, so that all patients should undergo life-long follow-up. As an example for a fulminant course of a malignant hemangiopericytoma (G3) we present the case of a 24-year-old female patient who initially presented nonspecific symptoms, was diagnosed late, and eventually died of her tumor despite chemotherapy and surgical treatment. PMID- 9410629 TI - [Osteosynthesis after sagittal division of the mandible. Biomechanical stability of various methods in a pig mandibular model]. AB - Forty years after the introduction of sagittal split osteotomy for transposition of the mandible according to Obwegeser, very different procedures for osteosynthesis are still discussed and practised. In a simple biomechanical model in the porcine mandible, five different methods for osteosynthesis using metallic screws (titanium, cobalt-chromium-molybdenum alloy) and one using a polymer screw (polylactic acid-copolymer blend), as well as the use of miniplates, were studied with regard to the stability of the compound. The Kruskal-Wallis H-test (variance analysis by ranks) showed statistically highly significant differences (P = 0.00017) regarding maximum stability. Osteosynthesis by miniplates was very stable with regard to the maximum load (Fmax = 234 N +/- 47), but not so in terms of three-dimensional stability of the osteosynthesis itself. The highest stability of osteosynthesis with screws only (Fmax = 183 N +/- 65) was found for a 2.7-mm titanium screw in triangular geometry. The use of 2.7-mm cobalt-chromium molybdenum screws (Fmax = 173 N +/- 42) and 3.5-mm titanium screws (Fmax = 160 N +/- 76) did not make an statistical difference (P = 0.37). The mechanical values of 2.0-mm titanium screws in linear (Fmax = 113 N +/- 37) or triangular (Fmax = 136 N +/- 62) geometry and of 3.5-mm polylactic acid-copolymer blend screws (Fmax = 121 N +/- 33) did not differ statistically from each other (P = 0.75) but they did from the previous group (P = 0.019). In consideration of the low biting forces following sagittal split osteotomy, all tested procedures of osteosynthesis meet the mechanical requirements for clinical practice. PMID- 9410630 TI - [Axiographic functional parameters after dysgnathia operations with special reference to the position of the temporomandibular joint]. AB - This follow-up study was designed to identify the differences between the TMJ movements of individuals undergoing orthognathic surgery, with and without positioning of the mandibular proximal segment. A further aim was to assess the location of the terminal hinge axis before and after surgery regarding variations due to mandibular advancement or set back, respectively combined two jaw osteotomy. Mechanical axiographic tracings were documented in 14 patients undergoing orthognatic surgery without positioning of the mandibular proximal segment. The terminal hinge axis position was marked by a tattoo. After surgery two more axiographic tracings were recorded over a follow-up period of 24 months. Changes to the terminal hinge axis position were also documented. Condylar guidance angles showed no differences, whereas Bennett angles decreased by an average of 25%. Subjective or objective TMJ symptoms after surgery could be observed in 64% of the patients, compared to 50% before surgery. Considerable variations in the position of the terminal hinge axis were observed nine months after surgery, especially in the two jaw group. In Class II individuals the terminal hinge axis was found in a cranial position both after 9 and 24 months. Class III individuals with two jaw surgery also gained a long term cranial localisation, whereas Class III patients with mandibular setback only demonstrated a hinge axis position inferior and anterior compared to the preoperative reference. No statistically significant differences were apparent between the axiographic condylar movement parameters in the present group as compared to data published employing condylar positioning. PMID- 9410631 TI - [Microneural reconstruction after iatrogenic lesions of the lingual nerve and the inferior alveolar nerve. Critical evaluation]. AB - As microneural repair techniques of the sensory mandibular branches enter the third decade of their clinical use, there are but a few long-term investigations into the value of these procedures in the treatment of iatrogenic injury to the lingual (LN), inferior alveolar (IAN) or mental (MN) nerve. To establish the efficacy of microneural repair in lesions of the LN, IAN or MN with loss of continuity, the outcome of sensory recovery was evaluated in a series of 92 patients (LN: direct coaptation n = 39, coaptation + sural nerve grafting n = 23; IAN: direct coaptation n = 11 coaptation + sural nerve grafting n = 10; MN: direct coaptation n = 11). The minimum duration of follow-up was 14 months postoperatively. The persistent sensory deficit was assessed using standardized neurosensory testing and gustometric stimuli. In addition the patients answered a multiple-choice questionnaire containing a list of complaints. To obtain a numeric estimate for interindividual and intergroup comparison the information from clinical measurements and patient reports was condensed into a 'neurological score' and a 'complaint score', respectively. Furthermore, adequate items from both scores were combined to affirm or deny the return of sensory function in terms of protective and discriminative sensation. The overall results show a broad range of variation in the scores, sometimes reflecting severe degrees of persistent sensory impairment. The lowest scores, corresponding to the best regeneration, were found after direct coaptation of the LN, IAN and NM, but even the best results did not provide sensory recovery to a preinjury level. After direct coaptation of LN 69% of the patients exhibited protective sensation and 41% regained discriminative function. In contrast, LN grafting was ensued from restoration of protective function in 39% and discriminative function in 17% of the patients. More striking differences were found between coaptation and grafting of the IAN (IAN coaptation: 91% protective function; 18% discriminative function; IAN grafting: 60% protective function, 0% discriminative function). In the LN coaptation group low scores and improved taste perception were convincingly associated with short periods since injury (i.e. timing of repair). In conclusion, we feel there is sufficient justification to optimize the potential results of microneural repair by immediate (LN/MN) or early (IAN) reexposure of the injured site in order to clarify the precise nature of the underlying nerve damage and prevent delay, if patients present with complete loss of sensory function subsequent to dentoalveolar or oral surgery. However, clinical and electrophysiologic findings suggesting impairment or partial loss of sensory function are considered a contraindication to microneural intervention, in view of the limited prospects of sensory recovery after surgical repair. PMID- 9410632 TI - [Raising a radial flap with primary wound closure by prefabrication of split skin fascia flaps]. AB - A disadvantage of the radial forearm flap is the removal of skin from a functionally important and aesthetically exposed region. To minimize the donor site morbidity with this flap, we have thus far used a two-phase procedure for intraoral defect coverage in 15 patients: In a first step, a 0.5-mm split thickness skin graft is transplanted to the forearm fascia and settles there over a period of 2 weeks. In step two, the prefabricated fascial-split thickness skin graft can be raised with complete preservation of the forearm skin and microsurgically transferred like a conventional radial forearm flap. We have obtained the following results with this procedure: (1) All skin grafts took completely on the forearm fascia. (2) Prefabricated fascial-split thickness skin flaps could be raised like conventional radial forearm flaps. (3) The very thin and moldable flaps were excellently suited for intraoral lining and showed complication-free healing. We conclude that tension-free, primary closure of the donor site can be achieved with minimal aesthetic and functional impairment. PMID- 9410633 TI - [Postoperative pain after interventions in the area of the mouth-jaw-face]. AB - In order to establish effective postoperative analgesia we studied the incidence and significance of pain following maxillofacial surgery. The trial included 102 patients undergoing one of six different surgical procedures. Postoperative pain was assessed using a visual analogue scale (VAS) and the short form of the McGill Pain Questionnaire (SF-MPQ) up to the third postoperative day. Postoperative pain intensity was significantly correlated to operating time, the frequency of analgesic demand and the type of surgery (orthognathic surgery > TMJ surgery > osteosynthetic surgery > osteotomy of impacted third molars > tumor resection > removal of osteosynthetic materials). Patient's age, sex and ethnic origin did not significantly affect the severity of postoperative pain. PMID- 9410634 TI - [Basaloid squamous epithelial carcinoma (basaloid-squamous carcinoma) of the mouth floor. Differential diagnosis and response to neoadjuvant chemotherapy]. AB - The basaloid squamous cell carcinoma (BSCC) is a variant of squamous cell carcinoma, having histologically distinctive features and appearing in the oral cavity, upper respiratory tract and esophagus. Histological hallmarks are the presence of a basaloid component in intimate association with squamous cell carcinoma. The basaloid component is characterized by tightly packed nests of cells with scant cytoplasm and hyperchromic nuclei without visible nucleoli and an increased mitosis rate. Basaloid squamous cell carcinoma is said to have a higher malignant potential than common oral squamous cell carcinoma with an increased incidence of regional lymph-node metastases and distant metastases. Our finding of a Ki-67 index of 30% and the immunohistochemical demonstration of p53 protein speaks well for enhanced aggressive biological behavior. The differential diagnosis includes the adenoid cystic, mucoepidermoid, neuroendocrine, adenosquamous and conventional oral squamous cell carcinoma. Because of early dissemination, radical surgical treatment and additional radiation therapy are considered necessary. Our findings indicate that partial clinical and histological tumor regression occurs after systemic neoadjuvant chemotherapy. PMID- 9410636 TI - [Oral, facial and cranial manifestations of von Recklinghausen neurofibromatosis (NF)]. AB - 19 patients with neurofibromatosis (NF) type 1 were evaluated retrospectively. Many patients presented not only with orocraniofacial neurofibromas but also with cranial skeletal deformities--involving skullbase, skull, orbit, midface and mandibula--and ophthalmological and dental symptoms, as well as neurological disorders. Malignant transformation of NF was observed in 1 case. Severe manifestations of NF were associated with the plexiform growth type. Due to the nature of NF surgical treatment most often is only palliative. PMID- 9410635 TI - [Diagnosis and prognosis of salivary gland tumors. An interpretation of new revised WHO classification]. AB - An exact morphological classification of salivary gland tumours is necessary for international comparison of clinical tumour studies. The basis is formed by the TNM system for determining tumour stage ("staging") and the WHO classification as the underlying principle of identifying the pathohistological tumour state and cellular tumour differentiation ("grading"). The second, revised edition of the WHO classification of salivary gland tumours differs from the first in that the exact definition of a considerably greater number of tumour entities is given and in the consideration of additional factors concerning prognosis and therapy. In the present interpretation of salivary gland tumours, not only are solitary tumour entities defined, but new findings are also considered concerning immunohistochemical tumour markers, proliferation markers (Ki-67 resp. MIB 1, AgNORs, PC-NA), oncogenes and cell receptors as well as cytogenetic alterations as prognostic factors. In particular the new tumour entities of adenomas (myoepithelial adenoma, basal cell adenoma, canalicular adenoma) and carcinomas (acinic cell carcinoma, mucoepidermoid carcinoma, polymorphous low-grade adenocarcinoma, salivary duct carcinoma, myoepithelial carcinoma) are characterized. In addition, the tumour-like lesions and differential diagnostic aspects are mentioned and a general review about new prognostic factors is presented. PMID- 9410638 TI - [Hydrodissection as an alternative method in esthetic facial surgery. Technical contribution]. AB - Conventional techniques in facial esthetic surgery are based on skin and flap elevation by sharp surgical dissection. The hydrodissection technique of local anesthetic administration facilitates creation of an atraumatic field for subcutaneous flap preparation. Its application in face- and necklifting procedures is described. PMID- 9410637 TI - [Use of a fibula bone span in alveolar ridge augmentation. Outcome after 2 years]. AB - A preliminary report presenting the results of fibular strut grafting in the severely resorbed mandibular and maxillary region is presented. Thirteen patients were treated due to severe resorption of alveolar and basilar bone of 49 segments of the mandible and the maxilla. Two patients additionally had pathological fractures of the mandible. In 10 cases the strut graft was harvested by means of a new minimally invasive technique. After modelling the fibular bone it was fixed to the recipient site by miniscrews or implants. After a mean follow-up period of 20 months (max. 31, min. 11 months) a retrospective analysis of clinical and radiological findings was carried out. It showed that a mean augmentation of 16 mm was achieved. Compared to other studies the fibular strut graft was resorbed less, and due to the primary stability it could be used for the treatment of fractures of the mandible. No more than natural resorption was observed when the patients received their prostheses fixed to dental implants. PMID- 9410639 TI - [Outcome of Van der Meulen columellar extension in bilateral lip-jaw-palate clefts]. AB - A total of 42 patients were operated on for secondary columellar elongation at the Department for Cranio-maxillofacial Surgery in Essen, Germany, from 1990 to 1996. The procedures were carried out because of nasal deformity in bilateral cleft lips (n = 35) or unilateral cleft lips (n = 7). The well-established drawbacks of Millard's forked flap technique, especially the resulting unfavourable scars at the lip columellar angle and the unpredictable, more conspicuous scar formations within the upper lip gave rise to an intensive search for an alternative surgical technique in our clinic. Since 1995 we have undertaken 14 secondary columellar elongations by means of Van der Meulen's three dimensional Z-plasty at the alar rim. Avoiding reopening of the lip we were able to lengthen the columella and project the nasal tip forward between 3 mm and 8 mm in all patients without introducing additional visible scars. The procedure led to an improved contour of the nasal tip, the alar rims and the nostrils. Nevertheless, we observed secondary broadening and shortening of the elongated columella in two children. This complication could subsequently be avoided by postoperative application of a nostril splint. The results confirm the effectiveness of the described technique for moderate columellar lengthening. The postoperative application of nostril splints is mandatory. PMID- 9410640 TI - [Primary soft tissue coverage and specific after-care of endosseous implants in pre-irradiated orbits]. AB - After orbital exenteration and high- dose irradiation (60 Gy on average), 17 endosseous implants were placed periorbitally in 5 patients. No hyperbaric oxygen therapy was performed. All implants were primarily covered with regional or free flaps. After second-stage surgery and aesthetic rehabilitation with an external maxillofacial prosthesis, the pocket depth and implant stability (periotest) were checked, an occipitonasomental radiograph was taken and the soft tissue assessed in short recall intervals. When the implant was uncovered, osseointegration was stable. Within a follow-up period of 35 months, no fixation had failed. In two patients, peri-implant inflammation (microbiologically confirmed Staphylococcus aureus) occurred, which was clinically only determined by soft tissue oedema and rubor. The results demonstrate primary soft tissue covering as essential for non irritating implant osseointegration in the irradiated orbita. In extraoral implants the clinical estimation of the peri-implant soft tissue, including a microbiological examination, is required for early detection of peri-implant inflammation in order to avoid secondary implant failure. In contrast, periotest and pocket depth are not relevant in recognizing an ensuing peri-implant inflammation. PMID- 9410641 TI - [Early detection of threatened implant loss in tumor patients]. AB - After resection of an oropharyngeal tumor, 157 dental implants were placed in 17 irradiated (44 Gy on average) and 20 non-irradiated patients. Within a control period of 37 months, 15 implants had failed. The reason for implant failure was analyzed, whereby indicative parameters were revealed. Eleven implants in four irradiated and one non-irradiated oral cancer patients showed no primary osseointegration during the healing period because of mandible fracture, overloading or for unknown reasons. Four implants in one irradiated and three non irradiated oral-cancer patients were lost subsequently on average 39 months after second-stage surgery due to biomechanical overloading or bacterial infection. No osteoradionecrosis development due to implant failure was observed in irradiated patients. In all cases, peri-implant pocket depth, implant stability and peri implant bone resorption increased before definitive implant failure. Therefore, these findings seem to be useful as indicative parameters in the prediction of implant failure. PMID- 9410642 TI - [Suppurative abscess-forming mediastinitis after tooth extraction. Consequences for therapeutic approach]. AB - Purulent mediastinitis is a rare but serious complication of a descending odontogenic infection with a high mortality. Diagnosis is difficult and frequently delayed. Physical examination is often nondiagnostic, but may include pronounced edema of the neck and chest. CT scan is the single most important tool for early diagnosis. The treatment is always is surgical, in combination with an extremely high dose of combined antibiotics. Ultimately, we only could save our patient with this therapy. PMID- 9410643 TI - Alternative medicine courses taught at U.S. medical schools: an ongoing listing. PMID- 9410644 TI - Education opportunities in alternative/complementary medicine for nurses. PMID- 9410645 TI - Overview of bone marrow and peripheral blood stem cell transplantation. AB - Bone marrow transplantation (BMT) and peripheral blood stem cell transplantation (PBSCT) are treatments used with increasing frequency for a growing number of cancers. As technology develops, so, too, does the complexity of nursing care. In addition, as the number of patients who receive BMT or PBSCT increases, more and more nurses will be involved in their care. Knowledge of what problems to anticipate, comprehensive assessment, clear patient and family education, and strong emotional support provide the key to successful patient management. PMID- 9410646 TI - Ambulatory care after bone marrow or peripheral blood stem cell transplantation. AB - Close follow-up care for the patient who has received either a bone marrow or peripheral blood stem cell transplantation is essential for the first year after the transplantation. The patient can experience many side effects, such as compromised immune system, graft versus host disease, veno-occlusive disease, and infertility. The role of the clinic or homecare nurse is to provide thorough assessment so as to detect treatable problems early and to identify nontreatable problems that might be affecting the patient's quality of life. PMID- 9410648 TI - Only the best is acceptable. PMID- 9410647 TI - Superior vena cava thrombosis associated with a central venous access device: a case report. AB - A case report is presented that illustrates the signs, symptoms, and treatment of superior vena cava thrombosis (SVCT) related to a venous access device (VAD). SVCT is a rare complication associated with VADs; its symptoms can be very subtle, which makes diagnosis difficult. Knowing who is at risk, assessing these patients closely, and providing patient education will help to identify early complications. Because patients spend most of their time away from healthcare providers, education is needed to increase patients' awareness of the subtle signs and symptoms of SVCT. Patients need to know what to report to their oncology team members so that early treatment can be initiated. PMID- 9410649 TI - Bone metastases: Part I--Pathophysiology. AB - Many cancers (especially breast, prostate, and lung cancers) metastasize to the bone. The most frequent site of bone involvement is the axial skeleton (i.e., cranium, ribs, spine, and pelvis). The sequelae of bone metastases include pain, hypercalcemia, pathologic fractures, and spinal cord compression. As patients survive for longer periods, effective management of bone metastases becomes critical to maintaining or improving quality of life. Controlling pain, preventing fractures and oncologic emergencies, and promoting mobility and function are the outcomes of successful management. Use of a clinical algorithm can assist the nurse in identifying bone metastases and managing the clinical sequelae. Knowledge of the pathophysiology and the ability to assess bone metastases will contribute to the nurse's ability to manage the clinical problems and to improve the quality of life of patients with cancer. PMID- 9410650 TI - Bone metastases: Part II--Nursing management. AB - Nurses play a crucial role in identifying bone metastases and managing clinical sequelae, such as pain. Understanding the metastatic process is necessary for delivering effective nursing care. Part I of this article described the pathophysiology and assessment. Part II will provide an overview of the nursing management of the sequelae of bone metastases, including pain, pathologic fractures, spinal cord compression, hypercalcemia, and anemia. Risk factor identification can lead to prevention and early detection of these clinically significant problems. Clinical management of bone metastases will contribute to the nurse's ability to improve the quality of life of patients with cancer. PMID- 9410651 TI - Intravenous potassium predicament. AB - In 1991, United States Pharmacopeia (USP) required modification of the nomenclature and packaging standards for potassium chloride injection following reports of deaths that occurred due to inadvertent medication administration errors. The following article taken from USP Quality Review provides a brief history of the revised standards and a current Medication Errors Reporting (MER) Program database analysis of potassium chloride misadministrations. Also included is an errors table that compiles and abstracts all actual and potential errors of potassium chloride for injection concentrate that were received through August 1996. Because errors continue to be reported, USP Practitioners' Reporting Network would appreciate continual communication about the potential for error when administering potassium chloride for injection concentrate. Reprinting of this review is encouraged. A camera-ready format also is available by contacting USP Practitioners' Reporting Network at 800-487-7776. PMID- 9410652 TI - The right to know. PMID- 9410653 TI - Transfusion therapy: nursing implications. AB - Blood product transfusion therapy and its concomitant risks pose a special challenge for the oncology nurse. Knowledge of the pathophysiology of transfusion reactions, presenting symptoms, and treatment is necessary to safely monitor transfusions. Through meticulous nursing assessment, detection, and intervention, oncology nurses can prevent and/or minimize morbidity and mortality in patients receiving transfusion therapy. PMID- 9410654 TI - Vancomycin-resistant enterococcus. AB - Vancomycin-resistant enterococcus (VRE) rapidly is becoming a significant cause of nosocomial infections. VRE lives in the environment (e.g., bedside tables, side rails, door knobs) for five to seven days, making transmission difficult to eliminate. Patients who are immunosuppressed or are receiving multiple antibiotics are particularly susceptible to developing VRE infections. Nursing management is the cornerstone of prevention and treatment. Nurses need to identify patients at risk, maintain isolation, and educate patients, family members, and other healthcare team members. PMID- 9410656 TI - Delegation: an increasing part of nursing care. PMID- 9410657 TI - Dealing with downsizing. PMID- 9410655 TI - Preventing chemotherapy dose and schedule errors. AB - Verification of dose and schedule are critical steps in chemotherapy administration. Less than optimal dosing can result in reduced efficacy, and an overdose can cause profound toxicity and death. Use of a systematic format for reviewing chemotherapy orders reduces the potential for medication errors. This systematic approach should include using current weights for dosage calculations, verifying all calculations, obtaining physician clarification when needed, and limiting chemotherapy administration to certified nurses. Patient safety is enhanced by careful verification of chemotherapy dosages and schedules. PMID- 9410658 TI - Management of dermatologic complications of chronic graft versus host disease: a case study. AB - The purpose of this article is to illustrate a multidisciplinary team collaborative approach to the management of dermatologic complications related to chronic graft versus host disease (GVHD). GVHD is a complication of allogeneic bone marrow transplantation (BMT) and peripheral blood cell transplantation (BCT). The skin is affected in more than 80% of patients with chronic GVHD who received a BMT. Involvement can be limited or extensive and can seriously affect patient morbidity and mortality. Nursing care of the patient with chronic GVHD induces careful patient assessment, patient and family education, and interventions aimed at preventing or minimizing complications. A coordinated multidisciplinary team approach is essential for the successful management of chronic GVHD. Nurses play a critical role in coordinating the multiple disciplines involved in the care of patients with chronic GVHD. PMID- 9410659 TI - 38th International Conference on the Biochemistry of Lipids. Assisi, Italy, September 16-19, 1997. Abstracts. PMID- 9410661 TI - Current readings in nuclear medicine. PMID- 9410660 TI - The San Diego Conference. Nucleic Acid Technology: The Cutting Edge of Discovery. November 6-8, 1997. Abstracts. PMID- 9410662 TI - [Rehabilitation of peripheral nerve lesions]. AB - After reporting the pathophysiology of denervation the authors deal with the changes that affect nerves while going through reinnervation. A review of the drugs that may help healing and general care to be adopted in order to avoid sequential pathological phenomena related to paralysis are shown. Rehabilitative treatment is discussed, the Authors dwell upon rehabilitation of neurogenous bladder of patients with spina bifida and rehabilitation of patients with paralysis of brachial plexus. Facial paralysis has been discussed before dealing with benefits and complications of electrotherapy. The use of electromagnetotherapy and electromyography are shown with a rich photographic and bibliographic support. PMID- 9410663 TI - [Holter monitoring of the arterial pressure in the follow up of a group of aged patients with chronic uremia]. AB - The cardiovascular complication represents the principal effect of death in patients uraemics cronics. The hypertension is one of most cause; for this, the control of hypertension is one important objective in this patients. The introduction of dynamic monitoring permit to estimate the adeguatesse of hypertension control or the appearance of extradialitic hypotension. The dynamic monitoring permit to value the difference in two groups, young and elderly and to conform the hypertension therapy. PMID- 9410664 TI - [The role of MRI in the valuation of disorders involving the bone marrow]. AB - MR is the most accurate imaging technique in bone marrow evaluation since it allows to recognize and distinguish its red and yellow components. MR shows high sensitivity but low specificity. At present, its role in bone marrow diseases evaluation is that of depicting the extent of the disease process and monitoring the effects of treatment. PMID- 9410665 TI - The Postprandial State: How Glucose and Lipids May Affect Atherosclerosis and Cardiovascular Disease. Proceedings of a workshop. London, United Kingdom, 25 November 1996. PMID- 9410666 TI - [Ambulatory surgery--a failed experiment?]. PMID- 9410667 TI - ["Physicians without boundaries"--the name is a program]. PMID- 9410668 TI - [Protection from competition in public health law. Decision of the Federal Social Court 28 August 1996 (6 RKa 37/95)]. PMID- 9410669 TI - [From the legal position. Cost measures]. PMID- 9410670 TI - [Robert Koch Institute: Multiresistant staphylococci--a serious problem in hospital health]. PMID- 9410671 TI - [Chronic pancreatitis--still a surgical disease?]. PMID- 9410672 TI - [New aspects in the pathophysiology of chronic pancreatitis]. AB - Despite numerous experimental and clinical investigations there is no consensus on the evolution of chronic pancreatitis (CP). In the 1970s and 1980s, Sarles persistently emphasised the de novo evolution of CP due to pancreatolithiasis. In recent years, however, clinical and morphological studies have provided strong evidence for the initial proposal of acute pancreatitis progressing to chronic pancreatitis. The so-called necrosis-fibrosis-sequence theory is supported by immunohistochemical work suggesting that inflammatory mediators primarily contribute to tissue destruction and that infiltration of pancreatic nerves by immune cells is a pathogenetic factor for the generation of pain in CP. While Sarles postulates that acinar hypersecretion and an imbalance of pancreatic stone promoting and inhibiting factors trigger the evaluation of CP, the necrosis fibrosis-sequence theory also involves other pathomechanisms (e.g. changes in ductal permeability, ischemia, oxidative stress) which have been shown to cause (acute) pancreatic injury. Despite this unifying template, which also lessens the need to identify independent mechanisms for the pathogenesis of acute and chronic alcoholic pancreatitis, there are still open questions, e.g. on genetic factors that (like in hereditary CP) may explain the different susceptibility of the pancreas to injury and the individual immunological response. PMID- 9410673 TI - [Drainage operation as therapeutic principle of surgical organ saving treatment of chronic pancreatitis]. AB - Intraductal and intraparenchymal hypertension represent the rationale for surgical drainage procedures in the treatment of chronic pancreatitis. "Simple" drainage procedures such as longitudinal pancreaticojejunostomy according to Partington-Rochelle have to be distinguished from "extended" drainage operations, e.g. the combination of longitudinal pancreaticojejunostomy with limited local excision of the pancreatic head. This "extended" drainage procedure according to Frey is just as effective as resective procedures in terms of persistent pain relief and definitive management of pancreatitis-associated complications of adjacent organs, i.e. distal common bile duct and duodenal stenosis. This operation also addresses an inflammatory mass in the pancreatic head. In contrast to "simple" drainage procedures the Frey operation allows reliable exclusion of pancreatic carcinoma. With low perioperative morbidity and zero mortality the Frey procedure significantly improves quality of life and leads to social and occupational rehabilitation. PMID- 9410674 TI - [Duodenum-preserving pancreatic head resection--a standard method in chronic pancreatitis]. AB - In patients with chronic pancreatitis the inflammatory process in the pancreatic head is frequently the pacemaker of the disease. In these cases an inflammatory tumor develops which leads to local complications in half of the patients. Duodenum-preserving pancreatic head resection, contrary to procedures used in the past, offers the possibility to preserve stomach, duodenum, biliary tree, and the insulin secretory capacity. Duodenum-preserving pancreatic head resection is a subtotal resection of the pancreatic head. In a series of 380 patients the hospital mortality rate was 0.8%, the frequency of reoperation 5.3%, and the median hospitalisation time 13.9 days. The early postoperative glucose metabolism was deteriorated in 2% and improved in 9% of cases. After a median follow-up time of 6 years, 88% of the patients were completely painfree or suffered pain rarely. Sixty-three percent were gainfully employed; the late mortality was 8.9%. Only 10% of the patients had further bouts of pancreatitis. The decisive advantage of duodenum-preserving pancreatic head resection over Kausch-Whipple resection is preservation of the endocrine pancreatic function and of neighbouring organs. PMID- 9410675 TI - [Pancreatic pseudocysts in chronic pancreatitis. Differential diagnosis and therapy]. AB - Pancreatic pseudocysts occur as a complication in the natural course of acute or chronic pancreatitis, with a spontaneous resolution rate of 40% to 60%. The incidence of pseudocysts in the course of chronic pancreatitis amounts to 60%. Pseudocysts have to be differentiated from neoplastic or congenital pancreatic cystic lesions. Careful diagnostic work-up including ERCP and CT scanning is mandatory in the management of pancreatic pseudocysts. The indication for treatment depends on the development of symptoms or complications. Pain is present in up to 90% of all patients, and the rate of complications varies between 2% and 55%. Pancreatic pseudocysts are usually an epiphenomenon of chronic pancreatitis. Pancreatic resection therefore represents a causative treatment. New percutaneous and endoscopic techniques for draining pseudocysts are challenging the previous hegemony of the surgeon. However, so far no prospectively collected data are available comparing the results of interventional and surgical drainage procedures. Therefore, in most symptomatic cases surgery is considered the treatment of choice. PMID- 9410676 TI - [Specializing in surgery. A contribution from the American viewpoint]. PMID- 9410677 TI - [Intraoperative cholangiography as a routine method? A prospective, controlled, randomized study]. AB - A prospective, controlled, randomized trial was conducted in 275 patients with symptomatic gall stone disease, whose history, laboratory data or sonographical findings did not suggest common bile duct stones. Of these patients, 137 did not undergo intraoperative fluoroscopic cholangiography (IOC), but in the remaining 138 patients IOC was attempted. In 111 cases (80.4%) the biliary system was sufficiently visualized. In 3 patients (2.7%) calculi in the cystic or common bile duct were diagnosed, which would have been overlooked without IOC. IOC was false-positive in one case. One year after the operation the patients were asked to return for a follow-up examination. Three patients in the group without IOC had had symptomatic passage of a stone, and one had a common bile duct stone removed by endoscopic papillotomy. A retained stone was discussed as etiology for a pancreatitis in a fifth patient in this group. No patient sustained long-term sequelae from the retained common bile duct stones. None of the patients in the IOC group had evidence of cholangiolithiasis at follow-up. There was no difference between the study groups concerning the incidence of post-operative complications. The operations with IOC lasted significantly longer (92 +/- 31 min vs 77 +/- 28 min). According to our data and those published earlier, the additional financial and logistic expenditure associated with routine IOC is not justified. Patients with the preoperative suspicion of a common bile duct stone should have endoscopic bile duct clearance (ERCP and EPT) prior to cholecystectomy. PMID- 9410678 TI - [Postoperative and post-traumatic acute cholecystitis. Value of ultrasound diagnosis]. AB - An often unrecognized but potentially fatal complication, mostly seen in posttraumatic patients under intensive care, is reactive acute cholecystitis. On account of the high specificity of ultrasound diagnosis in the biliary system we decided to examine the ultrasound criteria for early detection of posttraumatic cholecystitis. Ultrasound of the abdomen was performed prospectively, seven times on different days, in each of 40 artificially respirated patients under intensive care conditions over a period of 12 months. The results show that artificial respiration, parenteral feeding and previous trauma can lead to tardive (28/40) wall-thickening or to a three-layered wall of the gallbladder (9/40). In 22.5% of patients (9/40) we found the sonographic signs of acute cholecystitis. In correlation with the clinical signs, cholecystectomy was indicated in only two patients. The preoperative ultrasonographic findings and clinical signs of 23 patients with the diagnosis of acute reactive cholecystitis were analysed retrospectively. We found good correlation between sonographic and clinical signs of acute cholecystitis in 21 of these 23 patients. Our study shows that the morphological correlate of a thickened three-layered gallbladder wall can occur in the context of systemic alterations, even if there is no underlying cholecystitis. The diagnosis of acute reactive cholecystitis and the indication for cholecystectomy should be based on the synopsis of pathologic and clinical findings. PMID- 9410679 TI - [Anterior spinal artery syndrome after thoraco-abdominal esophagus resection. A rare complication]. AB - We report a case of postoperative paraplegia resembling an anterior spinal artery syndrome after curative esophagectomy in a patient with carcinoma of esophagus and clinical stage III (UICC). Neurologic deficit was characterized by loss of sensibility at the level of T12/L1 together with paraparesis of both lower extremities. Furthermore, dissociated sensorimotor depletion at C6/C7 (right sided) and at T5 (left-sided) was noted. This severe complication was most probably caused by peeling of an arteriosclerotic plaque of the thoracic aorta due to preexisting advanced arteriosclerosis, leading to a partial occlusion of the great radicular artery of Adamkiewicz. Even though anterior spinal artery syndrome is a well-known problem in the operative management of thoracic aortic aneurysms, this complication has not previously been reported after esophagectomy. PMID- 9410680 TI - [Fundus rotation gastroplasty: fewer cervical suture failures after esophagectomy?]. AB - Cervical leakage, occurring on average in 20-50% of the patients, is one of the major causes of morbidity following oesophagectomy for cancer. We report on a new technique of gastroplasty, namely fundus rotation gastroplasty which was used in 53 patients. There were 49 patients with oesophageal cancer and 4 with benign lesions. Hospital mortality was 5.7% (3/53) and the leakage rate 7.5% (4/53). The advantages of fundus rotation gastroplasty over conventional gastroplasty are the better blood supply and the greater length of the gastric tube. Controlled clinical trials will be necessary to confirm the advantages of fundus rotation gastroplasty versus conventional gastroplasty. PMID- 9410681 TI - [Open approach or Veress needle in laparoscopic interventions? Results of a prospective randomized controlled study]. AB - In a prospective randomized study on 50 patients undergoing laparoscopic surgery, the safety and feasibility of open access laparoscopy was analyzed and compared to the closed Veress needle technique. Open access laparoscopic surgery was performed in half the time needed for the Veress needle technique with equal safety and without complications or technical disadvantages. Furthermore, open access offers economical advantages, as disposable trocars are no longer needed. Therefore the open access technique is recommended as the standard for laparoscopic operations. PMID- 9410683 TI - [Delayed complications after extrapleural pneumonolysis for lung tuberculosis]. AB - The epidemic spread of tuberculosis after World War II and the deficiency of appropriate antituberculotic drugs led to a renaissance of surgical procedure such as plombage thoracoplasty, initiated in 1891 by Tuffier. Especially in Germany the insertion of paraffin and polyethylene was used in order to achieve an extrapleural pneumothorax in order to collapse the tuberculous cavities in the upper lobes. Due to a high rate of early complications and the assumed cancerogenicity, in a considerable number of cases the material was removed soon after its deployment. In some cases with the filling remaining in place, 30-40 years later infections and/or neoplasms occurred. From 1985 to 1996 in two centers of thoracic surgery 13 patients underwent procedures for removal of filling material. The patients suffered from infections (n = 11), malignant lymphoma associated with infection of the plombage (n = 1) and bronchial carcinoma (n = 1). Technically, we performed the thoracoplasty described by Schede (n = 9). Schede's thoracoplasty in combination with a muscle flap repair (n = 1) or partial resection of the thoracic wall (n = 1), an empyemectomy (n = 1), and an en-bloc pleuropneumonectomy (n = 1). All patients suffered from multiple underlying diseases (COPD, coronary heart disease, diabetes mellitus). However, apart from beside two procedure related deaths (pulmonary embolism n = 1, pneumonia complicated by multi-organ failure n = 1) no other major complications were observed. The plombage material in the case of malignant lymphoma is probably carcinogenic in relation to the time of exposure and should be removed in all cases. PMID- 9410682 TI - [Assessment of regional and systemic toxicity of isolated hyperthermic extremity perfusion with tumor necrosis factor-alpha and melphalan]. AB - Following isolated limb perfusion (ILP) with TNF alpha and melphalan the damage to muscle tissue and its systemic consequences in terms of myoglobinemia and myoglobinuria as well as the activation of the cytokine cascade were investigated. We measured the compartmental pressure of the limb during and after perfusion and determined the serum changes of myoglobin, creatine kinase (CK), interleukin (IL)-6, IL-1, s-IL-2-receptor, TNF-receptor, and ICAM-1 levels. The compartmental pressure rose significantly during ILP and decreased after reperfusion. Following its course, the decision whether to perform a fasciotomy or not can be more reliably made. Serum myoglobin levels exceeded 200 times normal values and the increase occurred significantly earlier than that of CK, thus enabling judgement of the risk of renal failure (crush kidney syndrome). The elevation of serum IL-1 and IL-6 values correlated with the frequency of cardiopulmonary problems (hyperdynamic shock) and facilitated counter-maneuvers. Our data, although obtained from ILP with TNF alpha, could be used to monitor toxicity also when other drug regimens are administered. PMID- 9410684 TI - [Gas gangrene mediastinitis after Boerhaave syndrome]. AB - Mediastinitis caused by infection with Clostridium perfringens and spontaneous rupture of the esophagus are both life threatening conditions. The combination of these two entities led to septic multiorgan failure in a 38-year-old woman. The patient was treated successfully by esophagectomy and postoperative lavage through a partially open abdomen. The lack of information regarding emesis, the leading symptom of Boerhaave's syndrome, caused delayed diagnosis: the triad of emesis, severe epigastric pain and emphysema of the skin was not established until 30 h after the onset of symptoms. PMID- 9410685 TI - [Abrikossoff granular cell tumor: a rare tumor of the esophagus]. AB - In the present case of a granular cell tumor an abdomino-thoracal resection of the esophagus was performed in a 41-year-old patient after preoperative diagnosis had suggested esophageal cancer. Granular cell tumors (Abrikossoff's tumor) of the esophagus are rather rare tumors of neuroectodermal benign histogenesis. Diagnosis can be obtained by endoscopic and histological study and immunohistochemical forceps biopsy. Because of the peculiar histological feature of the tumor, it can be mistaken for an squamous cell carcinoma. PMID- 9410686 TI - [Persistent chylothorax after fracture of the 12th thoracic vertebrae]. AB - We report on a patient who suffered chylothorax 2 months after she had undergone internal fixation of a fracture of her 12th thoracic vertebral body. The pleural effusion was treated by insertion of a chest tube. The chylothorax was managed conservatively. The patient received protein-rich nutrition supplemented with medium-chain triglycerides. As the volume of chylous fluid drained from the pleura had not decreased after 2 weeks, the patient received total parenteral nutrition without any oral intake of calories. Chest X-rays documented the disappearance of the chylothorax. Reexpansion of the lungs was noted, and the costophrenic sinuses could be clearly visualised. PMID- 9410687 TI - [Hygiene in the surgical department--recommendations by the National Reference Center for Hospital Hygiene]. AB - Many hygienic procedures performed in operation units are not supported by scientific investigations. The following recommendations by the National Reference Center for Hospital Epidemiology, founded by the German Ministry of Health in 1996, are based on the scientific literature and separate necessary from less necessary and unnecessary procedures. PMID- 9410688 TI - [Antibiotic prophylaxis in trauma surgery]. PMID- 9410690 TI - [Principles of antibiotic prophylaxis in vascular surgery]. PMID- 9410689 TI - [Antibiotic prophylaxis in visceral surgery]. PMID- 9410691 TI - Proceedings of the symposium on health impacts of large releases of radionuclides. Saint Petersburg, Russia, 27-29 February 1996. PMID- 9410692 TI - [British guidelines for HIV therapy]. PMID- 9410693 TI - [The effect of a lower stimulation frequency on the AV synchronicity of VDD pacemakers]. AB - BACKGROUND AND OBJECTIVE: Implantation of a VDD pacemaker (ventricular pacing; dual sensing [atrial and ventricular]; dual response [triggered + inhibited]) together with a single VDD electrode catheter restores synchronous AV ventricular stimulation in patients with higher-grade AV block and intact sinus function. If higher-frequency stimulation occurs it may be a sign of pacemaker malfunction or of inadequate pacemaker programming. This study was undertaken to determine, at first follow-up examination, in how many patients with a VDD pacemaker VVI stimulation occurred more than 5% of the time; how such patients differed from those with 5% or fewer VVI stimulations; and whether a changed program reduced the proportion of VVI stimulations. PATIENTS AND METHODS: 67 consecutive patients were tested 1 to 3 months after implantation of the Unity VDD pacemaker (Sulzer Intermedics). The frequency of VVI stimulations was determined via a diagnostic pacemaker memory store. After intermediate analysis, programming was optimized and the patients then re-tested 12 months after the initial implantation. RESULTS: At the first follow-up examination 54 patients had VVI stimulations of < or = 5% (0.5 +/- 0.9%) and 13 had > 5% of the time (19.8 +/- 10.7%). The two groups differed significantly from one another in their lower intervention frequency (< or = 5% VVI stimulations: 47 +/- 6/min; > 5% VVI stimulations: 58 +/ 5/min). In particular, the pacemakers in patients with > 5% VVI stimulations had been significantly more often programmed to values of > 50/min. As a result, the pacemakers of these patients were reprogrammed to a lower frequency. A year after implantation there was no longer any difference in the lower intervention frequency, 44 +/- 4/min, between patients with initially > 5% VVI stimulations and those with initially < or = 5% stimulations. At the same time, the proportion of VVI stimulations fell to 4 +/- 6%, with 67% of patients having AV synchronicity of > 95%. INTERPRETATION: At first follow-up, patients with > 5% VVI stimulations differed from those with < or = 5% stimulations with regard to an increased lower intervention frequency. In most of these patients the proportion of AV stimulations was increased to > 95% by reducing the lower intervention frequency to < or = 50/min. PMID- 9410694 TI - [Exposure testing with powdered gloves in 60 health care workers with latex allergy]. AB - BACKGROUND AND OBJECTIVE: Type 1 sensitization to natural rubber latex occurs in up to 22% of health care workers. Most sensitizations are due to the use of powdered latex gloves. Work place-associated exposure tests were performed to ascertain how frequently persons who developed breathing difficulties, rhinitis/conjunctivitis or obstructive respiratory tract illness when using powdered latex gloves are allergic to latex. PATIENTS AND METHODS: 60 persons (48 women and 12 men; mean age 29 +/- 7 years) with type 1 sensitization to natural rubber latex were studied. The past history was obtained through a questionnaire specially designed for those with latex allergy. Diagnostic measures included prick tests with different latex allergens, measuring the concentrations of total and latex-specific IgE and exposure trials, related to the work place, with powdered vinyl and latex gloves. RESULTS: The prick tests with various latex allergens produced significant skin reactions in 59 subjects, 58 had latex specific IgE antibodies (0.47 kU/l- > 100 kU/l). Exposure tests with powdered latex gloves produced rhinitis in 55 and conjunctivitis in 38 persons, urticaria in 3 and cough in 19. Whole-body plethysmography demonstrated a significant rise in respiratory tract resistance in 13, a reduction in 1-second expiratory volume of at least 20% in 15. Abnormal values in both tests were noted in 8 subjects. CONCLUSION: The breathing difficulty reported by all subjects was in most cases due not to bronchial obstruction but to obstructed nasal breathing. In cases with breathing difficulty on exposure to latex gloves a latex allergy should be considered as the cause. Powdered latex gloves present a danger to health care personnel and should no longer be used. PMID- 9410695 TI - [A delayed hemolytic transfusion reaction due to irregular antibodies with an anti-Kidd(a) specificity]. AB - HISTORY AND CLINICAL FINDINGS: A 56-year-old woman was admitted for replacement of a previously implanted right hip prosthesis which had become loose (no infection). She limped painfully on a shortened leg whose mobility was markedly impaired. At the site of the previous operation the pelvis was lower by about 2 cm; the scar looked well healed. INVESTIGATIONS: Routine laboratory tests were normal except for moderately raised erythrocyte sedimentation rate (18/ 27 mm). There was complete loosening of the prosthesis on X-ray. TREATMENT AND COURSE: Intraoperative bleeding from a branch of the inferior gluteal artery required blood transfusion and further erythrocyte infusions became necessary. Jaundice developed on the 4th postoperative day and 7 days later the direct Coombs test was positive with demonstrable agglutination. Free irregular erythrocytic anti Kidd(a) (anti-jk[a]) antibodies were found in the serum. To counteract the delayed haemolytic transfusion reaction, exclusively jk(a)-negative erythrocytes were infused. The jaundice gradually disappeared and the bilirubin values became normal. CONCLUSION: Jaundice and signs of haemolysis after erythrocyte transfusion may be due to delayed transfusion reaction and should be investigated with the direct Coombs test. PMID- 9410696 TI - [The rheological properties of hydroxyethyl starch]. PMID- 9410697 TI - [Kaposi sarcoma: a virus-induced tumor?]. PMID- 9410698 TI - [Oncogenes, differentiation markers and morphological criteria as prognostic parameters in non-small-cell bronchial carcinoma]. PMID- 9410699 TI - [Hemangioma diagnosis]. PMID- 9410700 TI - [Osteopathy treatment]. PMID- 9410701 TI - [Thromboses in inflammatory bowel diseases]. PMID- 9410702 TI - [The significance of risk-adapted antiviral prophylaxis and modern viral diagnosis for organ survival after kidney transplantation]. PMID- 9410703 TI - [Pregnancy in dialysis patients]. PMID- 9410704 TI - [The aortic arch as the source of a peripheral arterial embolism]. PMID- 9410705 TI - [Computer tomography-guided gastrostomy, jejunostomy and gastrojejunostomy. A reliable non-surgical method also for contraindications and failure of percutaneous endoscopic gastrostomy]. AB - OBJECTIVE: To analyse retrospectively the results of CT (computed tomography) guided gastrostomy or gastroenterostomy where an endoscopic procedure was not possible or had failed. PATIENTS AND METHODS: Between August 1993 and March 1997, CT-guided gastrostomy (n = 50) or gastroenterostomy (n = 8) was performed in 58 patients (mean age 56 +/- 6 years; 51 males, 7 females). An endoscopic or fluoroscopic method had been contraindicated in 52 and had failed in 6 patients. RESULTS: Technical success was achieved in all patients. Three patients needed intravenous sedation. There were no procedure related complications requiring treatment. Three patients had a mild infection at the site of the skin puncture. CONCLUSION: CT-guided gastrostomy or gastroenterostomy is a safe and simple procedure which provides a minimally invasive alternative also in patients with contraindications to the established percutaneous method. PMID- 9410706 TI - [Temporary and permanent pacemaker provision in persistent left superior vena cava and atresia of the right superior vena cava]. AB - HISTORY AND CLINICAL FINDINGS: An 80-year-old woman was admitted because of confusion episodes, difficulty of finding words and transitory clouding of consciousness on that day. There were moderately moist rales over both lung bases and mild pretibial oedema. The ECG showed atrial fibrillation with an irregular ventricular rate of 30/min. Attempted emergency implantation of a pacemaker electrode via the right subclavian vein failed because the catheter could not be advanced in the usual manner due to complete obstruction of unknown cause. INVESTIGATIONS: Fluoroscopy and venography of the upper great veins demonstrated a persistent left superior vena cava (LSVC), connecting to an aneurysmally enlarged coronary sinus (CS), as well as atresia of the right SVC (RSVC). TREATMENT AND COURSE: A temporary pacemaker was introduced transfemorally, subsequently replaced electively by a permanent one-chamber pacemaker system, the electrode wire having been introduced via the left cephalic vein--left SVC--CS, with its tip positioned in the right ventricle. CONCLUSION: If emergency placement of a pacemaker electrode proves impossible via the usual route because of a persistent LSVC and RSVC atresia, a transfemoral route should be used for temporary pacing. Permanent placement can usually be accomplished subsequently without much problem via the LSVC and coronary sinus. PMID- 9410708 TI - [Eosinophilia and the digestive tract. Eosinophilic gastroenteritis and its differential diagnosis]. PMID- 9410707 TI - [Ascites and suspected acute abdomen in hereditary angioedema due to C1 inhibitor deficiency]. AB - HISTORY AND CLINICAL FINDINGS: A 35-year-old man, for 8 years known to have hereditary angio-oedema with recurrent cutaneous swellings and occasional attacks of gastrointestinal pain, developed very painful, colic-like upper abdominal symptoms and frequent vomiting. INVESTIGATIONS: Routine laboratory tests were normal, except for leucocytosis of 18,200 WBC/microliter. The plasma concentrations of C1-esterase inhibitor (5.6 mg/dl) and of complement factor C4 (10.0 mg/dl) were reduced. Computed tomography revealed about 500 ml free fluid, a perihepatic effusion and definite oedematous thickening of the ileal wall. TREATMENT AND COURSE: During conservative treatment with infusions and no food by mouth the symptoms regressed. Abdominal ultrasonography was normal (no free intraabdominal fluid). Since discharge (now more than 15 months ago) the patient has been on danazole medication (200 mg/d). CONCLUSION: Recurrent gastrointestinal colics are typical of for hereditary angio-oedema and can imitate an acute abdomen. Concurrent ascites has only recently been described. Appropriate instruction of the patient and his/her medical practitioner is important to avoid unnecessary laparotomy. PMID- 9410709 TI - [An international comparison of the position of clinical research in Germany]. PMID- 9410710 TI - [The therapeutic potential of lazaroids (21-aminosteroids). A recent survey]. PMID- 9410711 TI - [Food incompatibilities]. PMID- 9410712 TI - [Early symptoms and anamnesis time in 517 patients with pituitary adenoma]. PMID- 9410713 TI - [Is transcatheter occlusion of a persistent foramen ovale a possibility for the avoidance of a paradoxical embolism?]. AB - BACKGROUND AND OBJECTIVE: Cerebral embolism may have different causes with sometimes serious consequences. If no specific reason can be found, paradoxical embolization through a persistent foramen ovale (PFO) is increasingly as a cause of the cerebral ischaemia. This study was undertaken to ascertain whether in patients with cerebral embolism occlusion of a PFO with a transcatheter technique can prevent further cerebral emboli. PATIENTS AND METHODS: Indications for transcatheter occlusion were based on neurological signs (ischaemic stroke), cardiovascular diagnosis, and coagulation tests. Between August 1991 and July 1996, transcatheter occlusion of a PFO was performed in 28 fully anticoagulated patients (median age 37.8 [15.4-65.4] years). The mean PFO diameter was 9.5 mm (3 17), mean duration of fluoroscopy 18.3 (8.7-43.1) min. The Rashkind device was implanted in three patients, the Sideris buttoned device in 25. During the follow up period (2-64 months; mean 13 months) renewed neurological symptoms occurred in only one patient. Transoesophageal echocardiography excluded thrombi on the implanted device or in the left atrium, and a residual PFO. The cause of the one neurological episode is therefore not clear. All other patients have remained free of symptoms and recurrence without anticoagulation after placement of the device. CONCLUSIONS: Transcatheter occlusion of a PFO is a relatively simple and safe procedure. Our results suggest that it can at least lower the incidence of further cerebral embolizations. The clinical value of the method in comparison with anticoagulation requires further study. PMID- 9410714 TI - [Chlamydia pneumoniae infection in a family]. AB - HISTORY AND SYMPTOMS: For 11 weeks a 38-year-old woman had suffered from a respiratory infection with peribronchitis, nocturnal coughing fits and earache. INVESTIGATIONS, TREATMENT AND COURSE: The Chlamydia-CFR titre was raised. Subsequent throat swabs of her husband and two daughters grew Chlamydia pneumoniae (C.p.), but not in her case; 5 days earlier she had been started on roxithromycin. 3 weeks before the patient fell ill her two daughters had a flu like illness with cough and subfebrile temperature and her husband also had flu like symptoms, which regressed after few days. Antibiotic treatment with roxithromycin improved the symptoms in the mother and older daughter, but the younger daughter was not given treatment because she had no symptoms at the time the microorganism had been isolated. She developed joint symptoms, like those of reactive arthritis, 12 weeks after the mother's illness had begun, and conjunctivitis 5 weeks later. CONCLUSIONS: It is likely that the daughters had the C.p. infection first and then infected their parents. While the father's and older daughter's illness quickly regressed, the mother became quite ill. Her serology was positive for a primary infection in adulthood, but in the daughters the serology was negative and, despite demonstration of the organism, the diagnosis of acute C.p. infection could not be made. PMID- 9410715 TI - [Anogenital condylomata acuminata, mucocutaneous candidiasis, vitiligo, keratopathy and primary hypoparathyroidism in an autoimmune polyglandular syndrome type 1]. AB - HISTORY: 6 months before admission a 21-year-old woman had developed anogenital condylomata acuminata (CA). Since early childhood she had been treated for primary hypoparathyroidism (PHPT) and recurrent mucocutaneous candidiasis. 5 years before admission corneal clouding had caused visual impairment. Recently, mainly truncal vitiligo with occipital poliosis had developed. INVESTIGATIONS: PHPT was confirmed (parathormone level 7.7 mg/l). In addition, liver transaminases were raised (GOT 105.8 U/l, GPT 145.6 U/l, gamma-GT 56.8 U/l), pointing to noninfectious hepatitis. An ACTH stress test could not exclude manifest adrenocortical insufficiency, and thyroid function was also normal. The Merieux Multitest indicated an anergy. DIAGNOSIS, TREATMENT AND COURSE: The constellation of test results suggested autoimmune polyglandular syndrome type I. The condylomata were treated by electrocautery and the intestinal candidiasis with amphotericin B suspension. Calcitriol capsules, 0.5 microgram, and calcium gluconate or lactate, 500 and 300 mg respectively, 3 times daily each, were given for the PHPT. CONCLUSION: This case demonstrates a complex syndrome which can be recognized early by simple clinical tests. Early diagnosis prevents possible life threatening complications. PMID- 9410716 TI - [Indications for heart transplantation]. PMID- 9410717 TI - [Hypothyroidism, hyperthyroidism and therapy with thyroid hormones: effect on the skeletal system]. PMID- 9410718 TI - [The problem of organ donation from the living]. PMID- 9410719 TI - [The problems of BSE-contaminated food and other products from beef]. PMID- 9410720 TI - [Merkel cell tumor of the skin]. PMID- 9410722 TI - [Effect of normal parturition on activity of clinically relevant enzymes in blood plasma of goats and kids]. AB - Blood of 24 healthy goats of different breeds in the age between two and six years as well as their 36 lambs in the peripartal period was taken at regular intervals from the day 10 ante partum up to the day 30 post partum. The content of plasma--nonspecific, clinical essential enzymes as well as the selenium content--has been examined. 1. Normal parturition shows an influence on the enzyme activity in plasma. Abundant alterations in concentration (p < or = 0.05 0.001) are detectable in AST, LDH, alpha-HBDH, GGT, GLDH and CK. GGT responds indistinctly to birth. GLDH-concentration is significant higher in the lactation period (p < or = 0.01). 2. The registration of OCT in blood plasma and serum of goats is possible but the activity of the enzyme in plasma and serum is slight. 3. In the blood of new born lambs a high plasma activity of CK and GGT can be found. Up to day 5 post natum the activity of CK descends to post partum adult level and up to day 30 post natum lower enzyme values than those of adult goats are found. GGT activity at day of birth after colostrum uptake is more than 5 times higher than at day 5 post natum. Up to day 30 post natum the activity remains above the mean of adult goats. PMID- 9410721 TI - [Clinical and microbiological investigations of atrophic rhinitis of Upper Austrian swine herds]. AB - The aim of this study was to determine the distribution of the progressive and non-progressive atrophic rhinitis of Upper Austrian swine herds. Further on the resistance pattern of the pathogens involved to chemotherapeutics was tested. In the period of May 1993 to June 1996 a total of 56 Upper Austrian swine herds were examined and on the occasion of the animal herd health management 997 nasal swab samples of young pigs taken. The area of this investigation included 14 Upper Austrian districts and the herds examined were divided into 3 types. Type 1 were swine herds of the swine breeding association (SZV), type 2 piglet producing farms (FP) and type 3 closed swine herds (GB). Sucking- and weaning piglets aged from 4 to 10 weeks were selected for these examinations. On the average 10 nasal swab-samples (2 swabs per animal) per herd were taken, microbiologically examined and the toxin by means of ELISA-tests determined. In terms of resistance testing (antibiogram) 20 different chemotherapeutics, which consisted of 11 different groups of drugs, were used. As a result of 997 nasal swab-samples examined, 304 (30.5%) P. m., 111 (11.1%) tox. P. m. and 35 (3.5%) B. b. were isolated. 50% of the Upper Austrian swine herds showed PAR by means of microbiological examination and ELISA-tests as well. The resistance pattern of P. m. and B. b. exhibited significant differences. Penicillin and lincospectin were highly resistant concerning isolates of B. b., but were highly sensitive for P. m. Enrofloxacin turned out as the most effective drug to the P. m.-toxin-negative- and B. b. strains tested, because no resistance was observed. Finally efficient PAR control programmes of swine herds belonging to the Upper Austrian swine herd health service are described. PMID- 9410723 TI - [Effect of insulin on glucose and and fat metabolism in ewes during various reproductive states in normal and hypocalcemia]. AB - Metabolic indicators of glucose and lipid metabolism, i.e. glucose turnover, insulin concentration in plasma, insulin clearance, concentrations of non esterified fatty acids (NEFA), glycerol and potassium were investigated in nine ewes during three reproductive states in order to examine their importance for development of subclinical ketosis. The increase of insulin in plasma was measured after a continuous 60 min intravenous infusion of glucose (4.9 mmol.min 1). Turnover of glucose and insulin clearance were quantified during a combined euglycemic, hyperinsulinemic clamp. Insulin was consecutively infused in doses of 5 and 10 mU.kg-1.min-1 for about 2 1/2 hours, each. Plasma glucose concentration was adjusted to 5.3 to 5.8 mmol.l-1. The experiments were carried out during non pregnancy and non-lactation, 4 weeks to 3 days before lambing and 3 to 4 weeks after lambing, each during normo- and hypocalcemia. Hypocalcemia (0.9 to 1.0 mmol Ca2+.l-1) was induced by continuous i.v. infusion of a 5% Na-EDTA solution. Infusion rate was continuously adjusted. The glucose induced increase in plasma insulin concentration was significantly lower during late pregnancy compared to peak lactation and non-pregnancy (46.3, 62.4 and 128 mU.l-1, respectively). The insulin clearance during a hyperinsulinemic clamp with 5 mU.kg-1.min-1 was significantly less during late pregnancy compared to peak lactation and non pregnancy (3.7, 6.0, 4.8 ml.kg-1.min-1, respectively). The concentrations of NEFA and glycerol in plasma during the infusion of 5 mU insulin.kg-1.min-1 were significantly higher during late pregnancy than during non-pregnancy (NEFA: 0.41, 0.04 mmol.l-1; glycerol: 96, 29 mumol.l-1, respectively). The results showed that insulin responsiveness was significantly reduced in sheep during late pregnancy. The insulin-mediated uptake of glucose by muscle and fat tissues and the insulin mediated inhibition of lipolysis were significantly reduced during late pregnancy compared to non-pregnancy and lactation. The diminished responsiveness of target tissue towards insulin during late pregnancy predisposed the animals for development of subclinical ketosis. Hypocalcemia exaggerated this situation by its inhibitory effect on hepatic gluconeogenesis and by enhancing insulin resistance of target tissues. The factors which are responsible for the altered responsiveness of target tissues towards insulin during late pregnancy are yet unknown. The potassium concentration in plasma showed a proportional increase with increase of the energy deficit of the target tissues. This effect could have been exerted by a decrease in cellular concentration of ATP and a concomitant reduction of the activity of Na(+)-K(+)-ATPase. The indicators of glucose and lipid metabolism which were examined in this study showed marked individual variation, particularly during late pregnancy. The marked changes of these indicators with reproductive stages as well as their great variation between individual sheep support the assumption that they are of significance for the development of pregnancy toxemia in sheep. PMID- 9410724 TI - [Effect of flunixin meglumine on plasma prostanoid concentrations in horses with colic in the perioperative period]. AB - In the present study the significance of eicosanoids in the development of shock in horses on the basis of ileus has been investigated using the prostanoids thromboxane B2 (TXB2) and prostaglandine E2 (PGE2) as indicators. The prostanoid synthesis inhibitor flunixin meglumine was to be examined regarding its efficacy in the effective blockade of the synthesis of these mediators within the peri operative timeframe as well as its effects on clinical signs and laboratory parameters. 21 horses suffering from ileus and ready for surgical intervention received an intravenous flunixin dosis of 1.1 mg/kg body weight immediately after the initial examination and prior to the surgical procedure. 20 colic horses receiving surgical treatment without application of the drug served as control group. Reference data concerning the approximate standard plasma levels of the prostanoids were determined in 10 healthy horses. Plasma levels of thromboxane B2 and prostaglandine E2 in all colic horses, treatment group as well as controls, initially proved to be significantly higher than the reference values in healthy horses. The untreated control group showed plasma levels highly exceeding the standards within the course of investigation. The application of flunixin meglumine resulted in an effective inhibition of the prostanoid synthesis. Post operatively as well as within the whole period of investigation the plasma levels of PGE2 and TXB2 of the treated group were considerably lower than those of the control group. Flunixin meglumine had a favorable effect on several cardiovascular parameters. The experimental data concerning the effects of flunixin meglumine thus could be validated in a clinical setting, especially the effective inhibition of the cyclooxygenase enzyme system. The application of the prostanoid synthesis inhibitor flunixin meglumine can be judged as being effective in limiting shock progress in the peri-operative setting given reliable diagnosis. PMID- 9410725 TI - [Clinical lung function investigations in sheep]. AB - In 144 clinical healthy and 97 sick sheep pneumotachographic investigations were performed. In 119 sheep the diagnoses "healthy" based on adspection, auscultation and palpatory percussion. In 25 additional healthy and in the sick animals the diagnoses based on clinical, endoscopical, serological, parasitological, and in 82 animals also pathomorphological findings. According to these diagnoses the animals were divided into the following groups: G = healthy, K = catarrhal purulent bronchopneumonia, E = suppurative pneumonia, W = verminous pneumonia, M = Maedi, MKE = Maedi + catarrhal purulent or suppurative pneumonia, MAKE = Maedi + Adenomatosis + catarrhal purulent or suppurative pneumonia, AKE = Adenomatosis + catarrhal purulent or suppurative pneumonia, and FAKE = animals of the AKE group at an earlier date of investigation. In all sheep respiration rate, tidal and minute volumes, forced tidal volume, peak flow and resistance were measured with the portable pneumotachograph Flowscreen. Tidal and minute volumes and flow rates depended at rest as well as after forced ventilation essentially from the respiratory rate and from the body weight of the animals. The average individual coefficients of variation of repeated measurements of healthy animals were between 21% and 36%. Only in tidal volume after forced ventilation (10%) and in the resistance (18%) the individual coefficients of variations were lower. In 57% of the animals with Lung Adenomatosis resistance-values over 0.4 kPa*s/l were measured, but in 94% of the sheep without adenomatosis resistance-values remained under this limit. PMID- 9410726 TI - [Bronchoalveolar lavage on pig breeding farms]. AB - In 182 pigs lung lavage was performed using a endotracheal tube and a catheter. The collected bronchoalveolar lavage fluid (BALF) was examined microbiologically. With decreasing numbers alpha-hamolytic Streptococci, Bordetella bronchiseptica, Haemophilus parasuis, Pasteurella multocida were cultured. Actinobacillus pleuropneumoniae was isolated from 3 BALFs. In one farm piglets were lavaged routinely for monitoring of the lung health status. PMID- 9410727 TI - [Involvement of the creatine kinase test in the selection scheme of pig breeding programs]. AB - With the increase of meat-quantity in the German pig-breeds in the sixties and seventies, more and more deficiency in meat-quality and stress appeared. It is based on a latent exertional myopathy. A result is an increase of Creatin-Kinase in the blood-plasma. It can be investigated by an analysis in a laboratory. The results from this analysis supplies us with information about the susceptibility to stress of the investigated pig. This physiological proceeding is consistently used in the CK-test to select stress-stable pigs. In HULSENBERG this test is used since the end of the seventies with great success. PMID- 9410728 TI - [In vivo investigations of stress susceptibility in pigs by means of magnetic resonance spectroscopy]. AB - 31P nuclear magnetic resonance spectroscopy was performed on 19 to 55 kg weighing pigs of different MHS genotypes to study the changes of phosphorus components (inorganic phosphate --Pi, phosphocreatine--PCr and adenosine triphosphate--ATP) of muscle metabolism as well as intramuscular pH under application of halothane. Aim of the present study was to observe the changes in energy metabolism and to perform a comparison with also measured blood parameters. Both, NN and Nn pigs did not show any changes during halothane exposure in phosphorus spectra, but in all animals a partially metabolically compensated respiratoric acidosis was found. In all MHS positive pigs a rapid fall of PCr and a corresponding raise of Pi levels in muscle was observed. PMID- 9410729 TI - [Epidemiological studies regarding the "summer infertility syndrome" of sows in confinement housing]. AB - In confined pig herds, located in Northwest-Germany and affiliated to a hybrid breeding organisation, 3 retrospective observational studies were performed to study the effects of "meteorological seasons" on sow fertility. A subsequent field trial was performed to study the effects of photoperiod and of temperature on sow fertility separately. Associations of economically important fertility disorders and season resp. climate categories were determined by adjusted Relative Risks in stratified analysis (observational studies), or by Odds Ratios in the Multiple Logistic Regression Analysis (field trial). Due to nonconsistent seasonal findings in the observational studies and a lack of temperature or photoperiod effects in the field trial we conclude that season and its main contributing climatic factors had no substantial effects on sow fertility in the well-managed herds of our climate zone. Nonclimatic risk factors for the so called "Summer Infertility Syndrome" are discussed. PMID- 9410730 TI - [Effects of vitamins A and E on the antioxidative metabolism of weaning pigs given dietary fats of different qualities]. AB - Early weaned piglets were divided into eight groups of 6 animals each. The animals were fed diets differing in fat quality (4% soybean oil, POZ 5 or 176) and in the content of the vitamins A (5,000 or 20,000 I.U./kg) and E (25 or 125 I.U./kg) over a period of 7 weeks. At the beginning, on day 25 and 47 blood samples were taken and analysed for vitamin A and E. In liver, heart, M. longissimus dorsi and M. semitendinosus vitamin A, E and the TBA-reactive substances were analysed. Induced lipid peroxidation was assessed by the ethane and pentane production rate in the skeletal muscle. During the weaning period a decrease in the alpha-tocopherol level was observed. In groups with the lower doses of vitamin E this effect was more pronounced. After 47 days the alpha tocopherol concentrations in plasma and heart and skeletal muscle fell about 25 30% by offering high doses of vitamin A compared to those groups fed low doses. Oxidized fats also led to lower tocopherol concentrations in muscle tissues. Hydrocarbon production in M. longissimus dorsi and M. semitendinosus was significantly reduced in groups with the high supplement of vitamin E. A tendentially opposite effect was seen in groups supplied with high levels of vitamin A or oxidized fat. Although retinyl esters in plasma are a minor fraction of the vitamin A activity, they present 99% of the vitamin A in the liver. The distribution pattern of the different retinyl esters was independent of the amount of supplementary vitamin A. In the present experiment 20,000 U of Vitamin A reduced plasma and tissue vitamin E levels. This effect led to an increase of lipid peroxidation indicated by the higher production of hydrocarbons. The results raise concerns about further increases of vitamin A supplementation in piglet feed. PMID- 9410731 TI - [Differential diagnosis of the porcine pituitary abscess syndrome (case report)]. AB - In a three years old sow, neurologic signs resulting from lesions of some cranial nerves and similar to the symptoms typical of porcine pituitary abscesses were described. A generalized leukosis with tumor cell infiltration occurring in numerous organs e.g. in the pituitary gland was diagnosed in this animal. The neurologic and pathogenetic differences between the described case and the porcine pituitary abscess syndrome were discussed. PMID- 9410732 TI - [The battle against Aujeszky's disease with the help of country-wide vaccination and the elimination of infected swine. III. Eradication of Aujeszky's disease in infected swine herds with the example of restoration proceedings in the area of Osnabruck/Lower Saxony]. AB - The eradication of Aujeszky's disease in an enzootic infected area of North-West Germany (Landkreis Osnabruck) is successfully treated by a combined vaccination, check and slaughter program as described. The important factors are the type of herd, herd size, pig density in the region and the completeness of vaccination which influence the eradication are also explained. To prevent future outbreaks of Aujeszky's disease in an area with high pig density it is necessary to vaccinate herds for at least two years after eradication. The importance of the further use of marker vaccines in a limited area around the outbreak even after stopping the area wide vaccination program is essential. PMID- 9410733 TI - [Motivation and proposals for revising the stock trading law]. AB - The German stock-trading law is a special statute that has been put into effect at the end of the last century (sections 482-492 BGB, i.e. civil code, and Kaiserliche Verordnung, i.e. imperial regulation). It promotes agricultural interests and contains guarantees which are out-of-date. This state had led to serious irritabilities with the general law of trade (sections 459-480 BGB). The difference in legal treatment of persons buying stock (i.e. equines, cattle, sheep and swine) or those buying other animals (e.g. dogs, cats, poultry) seems to be unsupportable any longer. For this reason the constitutional principle of equality in legal matters (section 3.1, German constitution) will be used and interpreted as a motivation for the importance to revise the antiquated stock trading law. PMID- 9410734 TI - [Toxicity of the combination of salinomycin and tiamulin in swine]. AB - The toxicity of the combination of salinomycin (sal.) and tiamulin (tia.) was investigated in dependence upon dosage and feeding method. In addition the efficacy of a safe dose for prophylactic treatment of dysentery was controlled. Following feed medications were tested for toxic effects in pigs: a) 3 mg sal. + 5 mg tia./kg BW, b) 3 mg sal. + 3 mg tia./kg BW, c) 3 mg sal. + 1 mg tia./kg BW, d) 3 mg sal./kg BW, e) 10 mg tia./kg BW, f) 30 mg tia./kg BW. The daily dose was given for 2 weeks by restricted feeding (twice a day) either as bolus or mixed in the whole ration or by feeding ad libitum. Animals were controlled for clinical symptoms and activities of creatine phosphokinase (CK) and aspartate aminotransferase (ASAT) were evaluated daily. Main clinical signs of poisoning were loss of appetite and locomotor disturbances and could be noticed for dosages of 8, 6 and 4 mg sal. + tia./kg BW. Activities of CK and ASAT were increased dose related, the feeding method also had an influence on the degree of intoxication. Some animals showed locomotor disturbances without any corresponding changes of CK and ASAT levels. Single pigs remaining without any symptoms even at high dosage pointed to differences in individual susceptibility. Toxicity was not found to be age dependent. Feed medication with 60 ppm sal. + 20 ppm tia. (feeding ad libitum) did not result in any signs of toxicity, however, the transmission of Serpulina hyodysenteriae from infected pigs to healthy, treated control animals could not be inhibited efficiently. Therefore the simultaneous application of salinomycin and tiamulin should be avoided generally, because the risk of intoxication is high and subtherapeutical dosage has an insufficient effectiveness against Serpulina hyodysenteriae. PMID- 9410736 TI - Haematological/Rheological and Neurological Aspects of Ischaemic Stroke. Proceedings of a satellite symposium at the 22nd International Joint Conference on Stroke and Cerebral Circulation. Anaheim, California, 8-9 February 1997. PMID- 9410735 TI - [Investigations on clinical diagnosis and pathogenesis of otitis media et interna in swine]. AB - A total of 25 pigs with a head tilt as clinical sign of otitis media et interna were examined. The majority were weaner-pigs with respiratory tract disorders. In lateral and ventrodorsal radiographic views of the bulla tympanica, there was an increased opacity, often accompanied by marginal destruction or thickening of the bulla wall. The findings confirmed the clinical diagnosis in each affected pig. In the case of leptomeningitis an examination of the cerebrospinal fluid revealed a drastic increase in the cell count. In 20 of 23 microbiologically examined empyemic bullae a polyinfection was seen. The results indicate that the route of infection of the middle ear is by the auditory tube. Mange on the other hand plays only a minor role in the pathogenesis of otitis media. PMID- 9410737 TI - Breast feeding: benefits and hazards. PMID- 9410738 TI - Interactive hormonal activity of chemical mixtures. PMID- 9410739 TI - Lead exposure and intelligence in 7-year-old children: the Yugoslavia Prospective Study. AB - For a prospective study of lead exposure and early development, we recruited pregnant women from a lead smelter town and from an unexposed town in Yugoslavia and followed their children through 7 years of age. In this paper we consider associations between lifetime lead exposure, estimated by the area under the blood lead (BPb) versus time curve (AUC7), and intelligence, with particular concern for identifying lead's behavioral signature. The Wechsler Intelligence Scales for Children-Version III (WISC-III) was administered to 309 7-year-old children, 261 of whom had complete data on intelligence, blood lead, and relevant sociodemographic covariates (i.e., Home Observation for the Measurement of the Environment (HOME), birth weight, gender, sibship size, and maternal age, ethnicity, intelligence, and education). These showed anticipated associations with 7-year intelligence, explaining 41-4% of the variance in Full Scale, Performance, and Verbal IQ. Before covariate adjustment, AUC7 was unrelated to intelligence; after adjustment, AUC7 explained a significant 2.8%-4.2% of the variance in IQ. After adjustment, a change in lifetime BPb from 10 to 30 micro/dl related to an estimated decrease of 4.3 Full Scale IQ points; estimated decreases for Verbal and Performance IQ were 3.4 and 4.5 points, respectively. AUC7 was significantly and negatively related to three WISC-III factor scores: Freedom from Distractibility, Perceptual Organization, and Verbal Comprehension; the association with Perceptual Organization was the strongest. Consistent with previous studies, the IQ/lead association is small relative to more powerful social factors. Findings offer support for lead's behavioral signature; perceptual-motor skills are significantly more sensitive to lead exposure than are the language-related aspects of intelligence. PMID- 9410741 TI - [Comparative microleakage study of two root canal sealants: calcium hydroxide and zinc oxide eugenol]. AB - A quantitative longitudinal study was carried out to compare the amount of microleakage using two different types of root canal sealers on straight and curved root canals. Microleakage was measured just after the setting of the sealer, one month and three months after that using fluid transport model. One month follow up results showed that in straight root canals both the ZOE containing Pulp Canal Sealer and the calciumhydroxide containing Sealapex sealers produced similar microleakage values to that values measured just after the setting of materials. One month follow up measurement resulted higher values in curved canals comparing to the straight ones in case of both the tested materials. Three months follow up measurement demonstrated worse results in the Sealapex groups than the Pulp Canal Sealer groups. With loglinear statistical analysis the following interactions were proved between variables: root canal form/microleakage, root canal form/time laps, and material/microleakage/time laps. Results of the present study are in good agreement with conclusions of recently published other studies, namely calciumhydroxide containing sealers resulted in higher microleakages than ZOE containing sealers. PMID- 9410740 TI - [The effect of certain stages of periodontal treatment on the regeneration of periodontal tissues]. AB - The regenerative potential of the periodontal tissues is relatively limited. The attachment loss has long been considered as an irreversible damage of the periodontium. Most of the conventional methods of the comprehensive periodontal treatment provided no convincing evidence of true new periodontal attachment formation. Most of the surgical and nonsurgical approaches achieved either secondary gingival recession and/or long epithelial attachement. The recently introduced guided tissue regeneration techniques can make the regeneration of the fibrous periodontal attachment and convincing clinical and histological evidences of new cementum and bone formation possible, as well as the regeneration of the perpendicular Sharpey's fibers fully embedded into the matrix of the appositionally formed new cementum and bone. The theories and clinical implications of these techniques are discussed and illustrated with clinical cases. PMID- 9410742 TI - [Incidence of injuries to deciduous teeth]. AB - An analysis of 191 primary incisors damaged by acute mechanical trauma on 108 children, ranging in age from 2 to 6 years, has showed that most of the patients were between 2 and 3 years of age (67%) when the accidents occurred. According to this clinical study the most frequent type of injury was partial luxation (42.93%). In 53.7% of cases the accident involved two teeth. The upper central primary incisors were damaged most frequently (82%). The ratio of boys to girls was 3:2. PMID- 9410743 TI - Newborn infants diagnosed as neurologically abnormal with relation to PCB and dioxin exposure and their thyroid-hormone status. PMID- 9410744 TI - Proceedings of the EASD/JDFI workshops. Oxford, United Kingdom, July 1995 and November 1996. PMID- 9410745 TI - [Effect of the shift and expedition-shift methods of labor activity on the hemostatic function of the body]. PMID- 9410746 TI - [Hyperkinetic states of hemodynamics in submariners]. PMID- 9410747 TI - [The psychophysiologic nature of intellect: in search of a working concept]. PMID- 9410748 TI - [Specifics of reorganization of brain rhythmic activity during perception of nonverbal information in children with speech pathology]. PMID- 9410749 TI - [Spectral analysis of electrical activity of the respiratory muscles and nerves]. PMID- 9410750 TI - [Oscillovasometric determination of the internal diameter of the major arteries of the extremities in man: comparison with the ultrasound method]. PMID- 9410751 TI - [Computer software for a visual search for evaluation of the effectiveness of selective attention]. PMID- 9410752 TI - [Dynamics of the psychophysiologic state of a commercial fisherman in the process of a long sail]. PMID- 9410753 TI - [Affinity of hemoglobin for oxygen and status of erythrocyte metabolism in athletes during intensive muscular work]. PMID- 9410755 TI - [Asymmetry of skin resistance response under activation of the right and left cerebral hemisphere]. PMID- 9410754 TI - [Principles of periodontitis therapy based on the correction of physiological defense mechanisms in the tooth area]. PMID- 9410756 TI - [Long-latency evoked auditory potentials in man during sequential paired clicks]. PMID- 9410757 TI - [Changes in brain bioelectric activity in liquidators of sequelae of the accident at Chernobyl power plant]. PMID- 9410758 TI - [Conditions of adequacy of the use of parameters of cardiac rhythm in evaluation of the psychophysiologic stress of operator activity]. PMID- 9410759 TI - [Characteristics of brain electric activity as parameters of the psychic state of parturients]. PMID- 9410760 TI - [Dynamics of brain electrical activity in 5- to 8-year-old normal children and children with learning difficulties]. PMID- 9410761 TI - [Individual minute-anxiety relationship in patients with neuroses]. PMID- 9410762 TI - [Memory disorders in patients with acute myocardial infarction]. PMID- 9410763 TI - [Pharmacologic evidence for disorders of myocardial adrenoreactivity in chronic coronary ischemia]. PMID- 9410764 TI - [Comparative analysis of sphygmograms of the carotid and radial arteries]. PMID- 9410765 TI - [Cardiohemodynamics in athletes with varying degrees of increased myocardial mass]. PMID- 9410766 TI - [Correlational relationship between parameters of thyroid hormones in newborn infants of mothers with and without hypothyrosis]. PMID- 9410767 TI - [Beta-adrenomimetic and beta-adrenomodulating properties of human urine]. PMID- 9410768 TI - [Endocrine regulation in aborigines and the immigrant population of the North]. PMID- 9410769 TI - [Hemodynamics in inhabitants of the "near" North]. PMID- 9410770 TI - [Transmyocardial revascularization with a holmium laser: preliminary results]. AB - BACKGROUND: Transmyocardial laser revascularization (TMLR) aims to improve perfusion of the ventricular wall via laser-created transmural channels. We present the results of TMLR with a holmium laser as sole therapy in patients with angina refractory to medical treatment and extensive coronary artery disease unsuitable for angioplasty or coronary artery by-pass grafting. METHODS: From November 1995 to February 1997, twenty-two patients underwent isolated TMLR with a holmium laser. Five patients (23%) were female; the mean age was 67 +/- 7 years (range 53 to 74 years). Previous myocardial revascularization procedures had been performed in 17 patients (77%). Mean preoperative angina class was 3.4 +/- 0.5 and unstable angina was present in 7 patients (32%). RESULTS: There were no hospital deaths. The only postoperative complications were transient supraventricular arrhythmias in 6 patients (27%). Each patient received a mean of 33 +/- 8 channels in 27 +/- 13 minutes. There were two late deaths, 40 days and 4 months after TMLR, due to stroke and myocardial infarction, respectively. Mean follow-up duration was 8 +/- 5 months (range 40 days-15 months). The mean number of hospitalizations due to angina fell from 4.9 +/- 1.5 in the 6 months before TMLR to 1.5 +/- 1.0 in the 6 months following surgery (p < 0.001). At follow-up, mean angina class had significantly improved (1.8 +/- 0.6, p < 0.001), as well as effort tolerance, which increased from a mean of 3.5 +/- 1.4 minutes to 5.1 +/- 1.7 minutes (p = 0.01). 201Tl SPECT at 3 and 6 months did not show any significant changes in the segmental perfusion of the lased and unlased areas. CONCLUSIONS: TMLR with a holmium laser is a simple procedure with low operative mortality and morbidity. Short-term results confirm that clinical improvement is obtained in most patients, although this is not supported by significant changes in myocardial perfusion at short-term follow-up. PMID- 9410771 TI - [Acute effects of ventricular and bicameral stimulation on plasma levels of natriuretic hormone]. AB - Atrial natriuretic factor (ANF) is a peptide produced by the atrium in response to increases in atrial pressure. It is a potent vasodilator and recent studies suggest that ANF may modulate vasomotor changes in patients (pts) with pacemaker (PM) syndrome. To evaluate the incidence of pacing mode on peptide secretion, plasma concentrations of ANF were determined in 32 pts (18 men and 14 women, mean age 71 +/- 4 years) with a DDD PM implant. Blood samples were collected one hour after a randomly assigned PM programming either in VVI or DDD mode at 70 ppm. Mean plasma ANF levels were 84.12 +/- 51 pg/ml in DDD mode and 156.0 +/- 15 pg/ml in VVI mode (p < 0.05). In 12 pts presenting ventriculoatrial retroconduction, the ANF levels were 77.16 +/- 50 pg/ml during DDD stimulation and 219.0 +/- 16 pg/ml during VVI stimulation (p < 0.05). ANF level was 88.50 +/- 46 pg/ml in DDD mode and 114.25 +/- 65 pg/ml in VVI mode in the 20 pts without AV retroconduction (p < 0.05). During DDD mode, 18 patients showed a DVI stimulation whereas 14 showed a VDD stimulation: the mean ANF level was 67.40 +/- 15 pg/ml during DVI and 100.40 +/- 28 pg/ml during VDD stimulation; the difference between these data was not significant. The increase in ANF levels during VVI pacing confirms the lower haemodynamic performance of this stimulation mode. The increase of ANF levels during VVI stimulation, which was in the subgroup without AV retroconduction, confirms the benefits of DDD stimulation also in this group of patients as well. Atrial pacing at physiological rates does not trigger the release of ANF. PMID- 9410772 TI - [Improvement of short-term prognosis in patients with refractory heart failure]. AB - BACKGROUND: Prognosis of patients with severe heart failure is poor, despite improved results in medical therapy. Heart transplantation is the only treatment possible in end-stage heart failure. The aim of this study was to evaluate the variation in prognosis over the past six years in the patients admitted to Intensive Care Unit for heart failure in spite of optimal oral therapy. STUDY POPULATION AND METHOD: Between January 1990 and December 1995, 133 patients with heart failure were admitted to the Intensive Care Unit, despite the fact that they were on optimal oral therapy. All patients were in New York Heart Association (NYHA) functional class III to IV and required intravenous administration of sympathomimetic amines, in addition to standard heart failure treatment procedures. Cumulative survival at six months of patients observed between 1990-1992 (group A) was compared with the survival rate of patients observed from 1993 to 1995 (group B). RESULTS: Clinical and haemodynamic parameters were similar in groups A and B, but ACE-inhibitors were used more frequently in group B (75 vs 31% respectively, p < 0.05). During the follow-up period, heart transplantation was indicated in a similar percentage of patients (A 53% vs B 58%). However, mortality on the waiting list (58% group A vs 21% group B; p < 0.05) and the percentage of patients who underwent heart transplantation (41% group A vs 78% group B; p < 0.05) differed. Moreover, all patients in group A and 50% of group B were operated on as "status one" patients. The total six-month mortality rate decreased from 69% before 1992 to 48% thereafter (p < 0.05). CONCLUSION: The short-term prognosis of patients with refractory heart failure improved over time. In the latter period, ACE-inhibitors were used more frequently and the number of heart transplantations was greater. Nevertheless, our results do not allow us to identify the causes of the improved survival rate. PMID- 9410774 TI - [I/D polymorphism of the ACE gene: a cardiovascular risk marker or error in molecular genetics?]. PMID- 9410773 TI - [Pharmacological cardioversion of isolated atrial fibrillation: recovery of atrial mechanical function and correlation with duration of arrhythmia]. AB - To evaluate how the duration of atrial fibrillation before cardioversion affects the recovery of atrial systolic function, serial transthoracic pulsed Doppler echocardiographic studies were performed within 2 hours and at 2 days, 7 days, 1 month and 2 months after chemical cardioversion to sinus rhythm. Peak A wave velocity (A), velocity time integral of A wave (A-VTI) and percent A-wave filling (A/VTI-tot) were assessed in 60 patients with lone atrial fibrillation of brief (> or = 12 to < or = 72 hours, 20 patients), moderate (> 72 hours to < or = 4 weeks, 20 patients) or prolonged (> 4 to < or = 24 weeks, 20 patients) duration. The three groups were well matched for age and left atrial size and none of the patients underwent antiarrhythmic therapy during follow-up. Atrial mechanical function is greater immediately and at 2 and 7 days after cardioversion in patients with brief atrial fibrillation, as compared moderate and prolonged atrial fibrillation (A and A/VTI-tot values: p < 0.05, p < 0.005 and p < 0.05, respectively). In addition, at 7 days after cardioversion, atrial systolic function is greater in patients with moderate atrial fibrillation as compared to prolonged atrial fibrillation (A value, p < 0.005 and A/VTI-tot value, p < 0.05). In all groups, atrial mechanical function increases over time and ultimately achieves similar levels. Full recovery of atrial mechanical function is achieved within 2 days in patients with brief atrial fibrillation, within 7 days in patients with atrial fibrillation of moderate duration and within 1 month in patients with prolonged atrial fibrillation. Recovery of the mechanical function of the left atrium is related to the duration of atrial fibrillation before cardioversion. These findings have important implications for assessing the early hemodynamic benefits of successful cardioversion. PMID- 9410775 TI - [Primary PTCA: reflections on organizational aspects]. PMID- 9410776 TI - [Angioplasty versus bypass in multiple-vessel coronary obstruction pathology]. PMID- 9410777 TI - [Co-management of heart transplant recipients: a three-way contract]. PMID- 9410778 TI - [Stable angina pectoris: what is the role of calcium antagonists?]. PMID- 9410779 TI - [Ergometric test and atropine: doubts about strategy]. PMID- 9410780 TI - [Fewer non-diagnostic tests]. PMID- 9410781 TI - [Four-year experience in health education in a sciences high school]. AB - Juvenile behaviour in front of health hazards is often based on irrational choices. In the attempt to form a prevention-oriented consciousness, ten years ago the University of Pavia and the Salvatore Maugeri Foundation organized an experimental prevention course in the local high schools. The intervention was effective and prompted the publication of a book describing the contributions, results and impressions of the experience. Utilizing this book as a teaching aid, the Authors have carried out a further four-year educative intervention, involving approximately 600 first- and second-year high school students. The lectures have illustrated the concepts of health, prevention and risk, and have briefly discussed the risk factors which are most relevant for the youth. The educative intervention has been performed with the cooperation of the school teachers. The answers to a questionary administered at the end of the course have confirmed that this kind of efforts promote a more rational behaviour aimed at health maintenance. The opportunity to introduce health education in the high school teaching programs has also been confirmed. PMID- 9410782 TI - [Evaluation of trends in occupational pathology in clinical cases]. AB - The study considered all patients admitted to the 1st Division of Occupational Medicine, Department of Preventive, Occupational and Community Medicine of the University of Pavia between January 1st 1989 and December 31st 1995 with a diagnosis of occupational disease. It was useful in illustrating the evolution of occupational disease as regards both changes in etiology and in the symptoms and systems involved. A decrease in the number of occupational diseases diagnosed from 1993 to 1995 was revealed, but it is difficult to draw any significant conclusions from these data since the need for hospitalization when diagnosing this type of disease has diminished greatly. Changes in legislation which have obliged employers to take precise preventive measures could also be partly responsible for this downward trend. The patients were, in almost all cases, at an initial stage of their disease. This is reassuring as far as the therapeutic and legal need for an early diagnosis is concerned. However, it remains unknown how often the causal relationship between work and disease was not suspected and therefore how many cases did not arrive to observation. Finally, the study confirmed the disappearance of some occupational diseases, such as benzene intoxication, common in the past, and the increase in "emerging" pathologies like allergies. PMID- 9410783 TI - [The occupational medicine physician and risk evaluation: between technical arguments and regulative obligations]. AB - Risk assessment (the characterization of potential adverse health effects of human exposure to environmental hazards) has been used by FDA and EPA for at least 30 yr, and many regulations currently rely on this procedure in USA and other european countries. RA method has been adopted for determinating of the exposure standards for relevant chemical and physical hazards. RA application shall become an important practice in Occupational Health and Industrial Hygiene, also in view of specific international and national regulations and recommendations. In this paper are discussed the main theoretical aspects of RA procedure in Occupational Health activities. Emphasis is given to the role of the Occupational Medicine Physician, who have a formal training and experience in toxicology epidemiology and industrial hygiene as well as in human physiology and clinical medicine. For these reasons the Occupational Medicine Physicians are in a unique position not only to give specific contributions in some steps of RA (mainly in risk identification and in risk characterization) but also to bridge the gaps between the various technical figures involved in the RA procedure and therefore to guarantee the adoption of correct and accurate preventive measures. PMID- 9410784 TI - [Cervical pain and proprioceptive sensitivity]. AB - With the intent of estimating the proprioceptive sensitivity of the cervical rachis, the Authors subjected 17 healthy volunteers and 20 patients suffering from various cervical disorders to a clinical head positioning test, which was previously experimented upon by several french Authors was modified under certain aspects by our equipe. The test, consists in flexion and extension of the head after maximal rotations to the left and right, then repositioning the head at the starting point "0" which corresponds to the crossing point of two orthogonal axis of a target. The positions are marked on the target by a laser beam situated on the helmet worn by the subject undergoing testing. The results obtained corresponding to the errors committed regarding point "0", permit us to confirm the reliability of the test and significant definitions between the control group and the patients suffering from cervical disorders. PMID- 9410785 TI - [Rehabilitation in multiple sclerosis: clinico-instrumental correlations]. AB - The Authors report their experience in a patient with Multiple Sclerosis. The planning and the follow up of rehabilitative treatment were based on clinical, functional, electrophysiological and posturographic evaluation. The impairment degree was evaluated by Kurtzke's Expanded Disability Status Scale. The electrophysiological evaluation showed an abnormal recruitment flexor pattern at exteroceptive stimulus and posturography confirmed postural control disorder. The treatment was based on PNF techniques and postural feedback on standing balance platform. After rehabilitative treatment the clinical and functional improvement has been confirmed by normal exteroceptive flexion reflex and better stabilometric parameters. PMID- 9410786 TI - [Clinical and EMG evaluation of the trapezius muscle in cerebrovascular disorders]. AB - A bilateral evaluation of the trapezius muscle was carried out, clinically and by surface electromyography, in 11 stroke patients, in order to compare the activation amplitude of the upper trapezius muscle in different movements of upper limb, using the healthy side as comparison parameter. At our request of analytical activation of the trapezius (consisting in raising their shoulders), we noted in all of them a contraction, both clinically and by EMG valuable. On the contrary at our request of the fixation activity of the shoulder girdle by the trapezius (humeral flexion) we couldn't notice a considerable activation in 4 patients. These data suggest that the trapezius is more active during voluntary movements than in postural fixation, and suggest the use of rehabilitation techniques focusing on the voluntary control. PMID- 9410787 TI - [Qualitative characteristics of human exposure to air chemical pollutants in operating rooms]. AB - For more than two decades many studies have been published searching for a link between exposure to volatile anaesthetic agents and health damage even if it is noteworthy that many other chemical substances can be found in the Operating Room. Purpose of this study was to demonstrate that the Operating Room is not a totally confined environment and that it is possible to perform an, at least qualitative, evaluation of many different polluting contaminants, even unexpected, to whom the working staff is exposed. MATERIAL AND METHODS: The study has been performed in the Operating Rooms of the Departments of Urology and Orthopaedics. Two methods have been employed: a long-casting sampling of volumes of air (with a sampling device composed of an enrichment system and a low flow aspirating pump) and an anaesthetic vapours and gas continuous analyzer. Results. We never recorded environmental levels of anesthetic higher than the currently accepted ones. Many other organic compounds of different kind have been found (irritants, cancer-organs). Their presence, not desirable in a place where a demanding work is performed, deserve further investigation and a quantitative evaluation of these compounds. PMID- 9410788 TI - [Indoor pollution and biological monitoring of volatile organic compounds (VOC)]. AB - Indoor air is a complex mixture of chemicals and airborne particles. Volatile Organic Compounds (VOC), a broad class of chemicals including diverse compounds such as Aldehydes, Terpenes, Aromatic and Aliphatic Hydrocarbons and Halogenated Volatile Organics, are an important category of indoor air pollutants. The evaluation of exposure to low doses of Chloroform and Benzene through the measurement of Chloroform and Benzene in urine was performed. Results show that biological monitoring may be helpful in indoor environmental studies in non occupational situations. PMID- 9410789 TI - [Hematuria in occupational medicine]. PMID- 9410790 TI - [Dyslipidemia associated with food]. AB - The increase of the morbi-mortality due to CHD in Mexico, particularly among the Social Security Institute (IMSS) workers led us to do research on the relative risk and the protection provided by foodstuffs usually consumed by these workers. We found significant evidence of low levels of cholesterol associated with dry alcoholic drinks, skimmed milk and yogurt as well as fresh cheeses. C-LDL was low among people that usually consume sweet alcoholic drinks and fresh cheese. High levels of TG were associated with those people consuming food products containing saturated fat (bacon, pork crackling, fatty red meat, fowl with skin) and viscera, more than three standard cups of alcoholic drinks three times a week, soft drinks and salt. Skimmed milk and yogurt and all vegetables were related to low levels of TG. Products related to high levels of C-HDL were all kinds of vegetables and beans. This study of IMSS worker eating habits could be useful to do research on the food intake of other worker populations, and could help us to design Health Education programs based on scientific knowledge. PMID- 9410791 TI - [Cysticerci-meningitis]. AB - Neurocysticercosis (NC) is the most common parasitic infection of the Central Nervous System (CNS) worldwide. Approximately 50% of patients with NC present cysticerci-meningitis; in spite of this, there is scanty information about this entity. This kind of presentation may lead to extremely serious disease and it often runs a chronic course. There is controversy about management, and some authors point out that use of steroids affords no benefits to the patient. We present 5 cases of cysticerci-meningitis with a follow-up of an average of 18 months, and we make important clinical considerations, underscoring the usefulness of steroids in the treatment of this serious condition. PMID- 9410793 TI - [The future of the Mexican Institute of Social Security. A view from inside]. PMID- 9410792 TI - [Glomerular hemodynamics in the isolated and perfused kidney. Effect of the inhibition of nitric oxide synthase]. AB - The isolated perfused kidney (IPK) is characterized by a normal glomerular filtration rate associated with a very low filtration fraction (FF). This study utilized micropuncture studies in the IPK to investigate: 1) the determinants of the ultrafiltration process responsible for the decrease in FF, and 2) the effect of nitric oxide synthase (NOS) blocker (L-NMMA) on glomerular hemodynamics. Two groups of kidneys were studied, a control group (CTL, n = 6) and an experimental group (EXP, n = 6) in which L-NMMA (0.56mM) was added to the perfusate. Significant differences in perfusate flow rate were required to maintain a constant perfusion pressure in both groups of kidneys. Nephron filtration rate, glomerular capillary hydrostatic pressure gradient and the ultrafiltration coefficient were similar between groups and identical to the ones obtained with in vivo micropuncture studies. Both afferent and efferent glomerular resistances were significantly reduced, although L-NMMA administration increased significantly both resistances. Altogether, these data demonstrate that 1) the IPK is an excellent preparation to evaluate the effect of different agents on glomerular pressure and/or membrane permeability in the absence of systemic effects; 2) the decrease in glomerular resistances leads to a significant increase in perfusate flow required to maintain glomerular pressure within normal range, and 3) in the absence of any systemic effect, NOS blocker increases both afferent and efferent arteriolar resistances. PMID- 9410794 TI - [Advances in the investigation of schizophrenia]. AB - The recent advances in the investigation of schizophrenia were reviewed. Three major fields are considered relevant: 1. Genetic studies with the remarkable finding of the 6pter-p22 chromosome linkage study, which will lead to better understanding about inheritance of disease in the future; 2. The new x-ray techniques, including our own investigations showing smaller brain size in our patients, and 3. The prominent new antipsychotic drugs, as effective as the older drugs but with fewer side effects. PMID- 9410795 TI - [Rheumatic diseases in colonial Mexico]. AB - The medical texts published in New Spain between the XVI and XVIII century have not been searched until now for evidence of the existence of rheumatoid arthritis and other rheumatic diseases before the descriptions made by Sydenham. We surveyed most of the medical books written and published in New Spain from the arrival of the Spaniards to the XVIII century, and we divided the diseases with articular manifestations into four groups, according to their main clinical characteristics: pain without swelling in anatomical region; pain without swelling in several joints; pain and swelling in joints, and joint complaints associated with contagious diseases. We found that a difference was established between gout and rheumatoid arthritis one hundred years before Sydenham, according to the different evolutions of both diseases, and we present one of the oldest descriptions of reactive arthritis. PMID- 9410796 TI - [Sequential treatment with granulocyte-colony stimulating factor (GCSF) and human recombinant erythropoietin (rH-EPO) in anemia of a patient with myelodysplastic syndrome and high blood transfusion requirements)]. AB - We describe the case of a patient with myelodysplastic syndrome (MDS) classified as Refractory Anemia with our Excess blasts, who suffered from high transfusional requirements and who did not respond to the administrations of B12 vitamin, folates, danazol, low dose cytarabine or recombinant human erythropoietin (rHuEPO). The patient was administered two cytokines: granulocyte colony stimulating factor (G-CSF) followed by rHuEPO. The patient remained transfusion free for more than 4 months until his death from causes not related to MDS or the therapy he received. It is the opinion of the authors that the initial G-CSF administration stimulated the early erythroid precursors, making them capable of finishing their maturation when rHuEPO was administered. We believe that this could be a useful therapeutic measure in the treatment of patients with MDS and high transfusional requirements. PMID- 9410797 TI - [Usefulness of computed tomography with double contrast media and delayed enhancement in multiple sclerosis]. PMID- 9410798 TI - [Serial beings. Cloning in mammals]. PMID- 9410799 TI - [Neurosteroids, GABA receptors and neuronal protection]. PMID- 9410800 TI - [Management of patients with pulmonary tuberculosis in a health care institution. A survey on nursing personnel knowledge]. PMID- 9410801 TI - [Abortion. Biological elements for legislative reflexion]. PMID- 9410803 TI - [Ectopic pregnancy]. PMID- 9410802 TI - [Resolutions of the International Society for Research on Civilization Diseases and the Environment, on the usefulness of chronobiology in the clinical evaluation of blood pressure]. PMID- 9410804 TI - [Risk factors during the intrauterine stage predisposing to congenital hip dislocation]. PMID- 9410805 TI - [Vaginal hysterectomy assisted by laparoscopy. Critical and comparative study if vaginal and abdominal hysterectomy at the A.B.C. Hospital of Mexico]. AB - The laparoscopically assisted vaginal hysterectomy offers the possibility to convert abdominal to vaginal approach give to the patient the benefits of ti. The analysis of the first 20 cases in our institution is done and are compared with abdominal and vaginal approaches in some parameters including costs, indications and hospitalization days. The laparoscopically assisted vaginal hysterectomy offers to the patient the benefits of the vaginal approach with hospital stay similar and cost and operative time higher than those for either vaginal and abdominal hysterectomy. The exact role of the laparoscopically assisted vaginal hysterectomy on daily practice still is pending. PMID- 9410807 TI - [Advanced maternal age and pregnancy: how much is too much?]. AB - Perinatal evolution was compared and two study groups in women with advanced maternal age and pregnancy. 626 were included from a total of 778 with age 35 years, who resolved their pregnancy during 1995. They were classified, according to age, in two groups: 1) maternal age of 35-39 year; they were considered primigestas and multigestas. Perinatal complications were classified in personal antecedent, antepartum and intrapartum complications. To analyze the association between maternal age and parity with perinatal complications, X2 or exact test of Fisher, was used. Percentage of women with advanced age and pregnancy was 13.6%. Main perinatal complications were: preeclapmsia, gestational diabetes, preterm delivery threat, and membranes rupture. There were no significant differences as to complications by age and parity. There were 90% of children with 2500 g, and Apgar of 97%. Perinatal death was 0.4%, and fetal malformation 0.6%. Cesarean frequency, was over 90% in primigestas and in more of 60% in multigestas. Perinatal evolution in advanced age and pregnancy is adequate, if she starts prenatal control early enough. PMID- 9410806 TI - [Study of the sterile male]. AB - The approach on evaluation of the infertile male must be the same used to evaluate other medial problems. It must be obtained a detailed interrogation with emphasis upon areas affecting infertility. Interrogation must be followed evaluation of infertility guide us to the identification of specific abnormalities responsible of infertility. However this is only possible in some cases, in others there are abnormalities of seminal count whose etiology can not be established with accuracy. PMID- 9410808 TI - [Cholestasis in pregnancy. Report of a case]. AB - We present a case of Cholestasis and pregnancy that developed associated in the puerperium, to tecaluteinic ovaries and hemoperitoneum. For this reason it was necessary surgical management. We considered that it is a very unusual case so we felt interest on presenting it. To finish we would to make reference to itns possible patogenia, symptoms, and management of it. PMID- 9410809 TI - [The proportion of molecular forms of gonadotropins changes in patients with polycystic ovaries]. AB - It is known that protein hormones circulate as different molecular forms and the relative proportion of these isoforms changes according to endocrine milieu. In particular gonadotropins, both LH and FSH, isoforms suffer variations related to the estrogen levels; thus sera obstained from menopausal women show a predominance of larger molecular forms which are considered as having lesser biological activity and the administration of estrogen replacement therapy is followed by the appearance of intermediate molecular forms possessing higher biological activity. This chormatographic pattern with predominance of intermediate isoforms is typical at midcycle in sera from normal women at the periovulatory stage. Present study showed that sera obtained from anovulatory women, such as patients with polycystic ovaries a predominance of larger and smaller molecular weight isoforms, exhibiting a chromatographic pattern different from that observed in normal women. It is speculate that there is some imbalance between the ovarian steroid synthesis and gonadotropin production in the stage of tertiary structural conformation. PMID- 9410810 TI - [Gynecologic surgery in geriatric patients]. AB - To establish a precise definition for the geriatric patient, results difficult, since the age limits change constantly and arbitrary way, but given that women are living longer, supposedly the rate of gynecologic surgery is increasing. OBJECTIVE: To inform the experience obteined in our service, from geriatric patients, with surgical intervention, and compare the results with published articles from other national gynecology services. MATERIALS AND METHODS: In the gynecology service of Specialties Hospital "Dr. Miguel Dorantes Mesa" from the S.S., in Xalapa Veracruz, Mexico, a retrospective study was performed, from 76 cases of patients with gynecologic surgery. The research variables were: 60 years old or older, personal pathologic data, gynecologic and obstetrics, occupation, preoperative diagnosis, type of intervention, anatomopathologic diagnosis, anesthesia employed, complications during of after surgery, days in the hospital, hemoglobin and hematocrit. RESULTS: Most of the patients were 65 to 70 years old, 100% housewives, pelvic statics alterations were present in most cases (53.9%), followed by pre-malignant and malignant diseases of the cervix, the abdominal hysterectomy was indicated in 29 cases, 65.7% had personal pathologic data, the anatomopathologic study confirmed 85% of the cases, days-hospital average was 5.8. CONCLUSION: Survivorship does not depend by type of surgery or age, if not to concomitant illness. PMID- 9410811 TI - [Aortic contraction in pregnancy: report of a case at the Central Military Hospital]. AB - The coarctation of the aorta is an uncommon pathology which has a high mortality rate during pregnancy; a very thorough antenatal control must be performed due to the serious complications that may cause death. It is important to eliminate any congenital malformations related to this pathology, such as aneurysms of the Willis polygon, Ductus Persistent or ventricular intercommunication; arterial pressure must be strictly controlled using antihypertensive drugs when needed with beta-blockers. The delivery route may be vaginal, C-section must be reserved only for obstetric reasons. We present a case studied in the Military Hospital, treated with antihypertensives, the fetus had intrauterine growth retardation and was delivered by C-section successfully. PMID- 9410812 TI - [Usefulness of ultrasonographic markers in chromosomal abnormalities]. AB - During a 3 and 1/2 years, 132 pregnancies were diagnosed as having a wide variety of congenital abnormalities. A high resolution ultrasound and multidisciplinary approach was used. In 95 cases fetal karyotyping was made. In this group the incidence of chromosomal abnormalities diagnosed during the period and phenotypic expression of the different types of chromosomal abnormalities was investigated. 29 abnormal karyotypes were found; 11 trisomy 18, 7 in monosomy X, 4 in trisomy 21, 3 in trisomy 13, 1 with tetraploidy (92XXYY), 1 Turner mosaic (45XO 68% 46XY 32%), 2 inversions of choromosome 9. Of the total abnormal chromosomal diagnosed during the period (N = 57), this group represented 49.2%, compared to 5 to 15% found in other risk groups. 224 congenital abnormalities were found. 43 (19%) isolated, and 181 (81%) associated. Of the 224 congenital abnormalities diagnosed, 80 (36%) were associated with chromosomal abnormalities. The most associated markers were duodenal atresia, heart defect, microcephaly, enlarged posterior fossa, and cystic hygromata. A specific markers pattern was found for each aneuploidy; heart defects for trisomy 18, holoprosencephaly and faciel cleft for trisomy 13, and cystic hygromata for monosomy X. It was concluded that the ultasound can be the most useful method to select the group of pregnant women with a higher risk of abnormal karyotype. PMID- 9410813 TI - The relationship between body size and mixed-species troops of tamarins (Saguinus spp.). AB - While Saguinus fuscicollis forms mixed-species troops with sympatric tamarin species in parts of its geographic range, in other parts it does not. The question is addressed whether body size divergence is a critical factor for this difference. Analysis of body size (head-body length) data shows that body size divergence ranges between 8 and 17% for associated species and between 1 and 4% for non-associated species. In associated species, the degree of body size divergence seems to correlate with the stability of mixed-species troops (i.e., time spent in interspecific association). It is concluded that body size plays an important role for niche differentiation, sympatric coexistence and the formation of mixed-species troops in tamarins. PMID- 9410814 TI - Self-suckling behaviour by a wild chimpanzee. PMID- 9410815 TI - [Pertussis vaccination with acellular vaccines. Tolerance--effectiveness--current vaccination recommendations]. AB - Pertussis is one of the most common infectious diseases in children, affecting in particular nonimmunized babies and young children, but increasingly also adolescents and adults. Complications occur for the most part in infants and in addition to infectious complications may also even lead to death from apnea. Since 1991, general pertussis vaccination has been recommended again, but because of the relatively high rate of side effects associated with the whole-cell vaccines available, has remained at a low level. This led to the development of acellular pertussis vaccines with appreciably improved tolerability. A number of these acellular vaccines offer good protection, and are approved for immunization. Owing to their excellent tolerability and the resulting better acceptance, acellular pertussis vaccines can considerably improve the immunization rate. Only in this way will it be possible to reduce the incidence of one of the most common infectious diseases of childhood that is also associated with the highest rate of complications. PMID- 9410816 TI - [Problems of whooping cough not yet solved. Epidemiology and vaccination strategy in Germany--country-wide field study]. PMID- 9410817 TI - [Consider whooping cough also in adults. Interview by Dr. rer. nat. Anita Schweiger]. PMID- 9410818 TI - [Chronic hepatitis. Differential diagnosis and therapy. 4: Diagnosis of autoimmune liver diseases]. PMID- 9410819 TI - [Slow release nifedipine in treatment of hypertension. 24-hour effect on blood pressure and heart rate--treatment follow-up of 391 patients]. PMID- 9410820 TI - [Gynecologic operations in fibromyalgia syndrome. A retrospective analysis of 890 patients of a rheumatologic and general practice]. PMID- 9410821 TI - [Drug problem should not be penalized. Interview by Dr. med. vet. Susanne Kammerer]. PMID- 9410822 TI - [Differential therapy of hypertension becomes more specific. Interview by Dr. R. G. Sommer]. PMID- 9410823 TI - [Good immunologic arguments. Reader opinion on immunologic origin of psoriasis vulgaris]. PMID- 9410825 TI - Proceedings of the XVth International Symposium on Endocrinology and Development. Paris, France, November 28-30, 1996. PMID- 9410824 TI - [Prediction: onset of improved mood after 2 weeks. Hypericum therapy in intermediate class depression. Interview by Dr. rer. nat. Till Uwe Keil]. PMID- 9410826 TI - Proceedings of the Workshop on Application of Biomarkers to Cancer Epidemiology. Lyon, France, 20-23 February 1996. PMID- 9410827 TI - Medical practitioners and their practices in acute diarrhea. PMID- 9410828 TI - [Long-term gastrointestinal monitoring--pH-metry and mobility]. PMID- 9410829 TI - [Long-term monitoring of respiratory function and blood gases]. PMID- 9410831 TI - [Long-term monitoring by score systems in intensive care medicine]. PMID- 9410830 TI - [Long-term monitoring in neurological diseases]. PMID- 9410832 TI - [Strategies for long-term registration of physiologic parameters in internal medicine]. PMID- 9410833 TI - [70-year-old patient with rapidly progressing dysphasia, disorientation and somnolence]. PMID- 9410834 TI - [Differential diagnosis of cavernous lung structures]. PMID- 9410835 TI - [Effect of upper arm circumference on RR measurement]. PMID- 9410836 TI - [Screening in activated protein C resistance]. PMID- 9410837 TI - [Therapy of transfusion hemochromatosis]. PMID- 9410838 TI - [Indications for acetazolamide]. PMID- 9410839 TI - [Prevention of para-pneumonic pleural adhesions]. PMID- 9410841 TI - [Polymyalgia rheumatica and giant cell arteritis]. PMID- 9410840 TI - [Systemic therapy with antimycotic agents]. PMID- 9410842 TI - Proceedings of the 10th Autoimmunity Meeting. Tel-Hashomer, Israel, 20 March 1996. PMID- 9410843 TI - [Back mutation mosaic: a new genetic principle]. PMID- 9410844 TI - [Telomerase]. AB - Telomerase is an enzyme that elongates telomeric repeats, the specialized structures at the ends of chromosomes that provide genomic stability and compensate for the physiologic process of telomere shortening. It has been implicated in cellular senescence, immortalization, and carcinogenesis. Over 85% of human tumours, and 95% of nonmelanocytic skin cancers, show telomerase activity, in contrast to the corresponding normal tissues. This suggests that telomerase activity may play an important role in carcinogenesis. Recent evidence shows that telomerase is active not only in embryonal and germ line tissues, but also in some normal tissues. In the skin, this activity has been traced to the stem-cell-bearing epidermal basal cell layer, possibly reflecting the presence of telomerase-competent stem cells. These findings require a reconsideration of our interpretation of telomerase activity in tumours of the skin and other tissues. As a causal relationship linking telomerase activity and cancer has yet not been demonstrated, some caution is warranted. PMID- 9410845 TI - [Classification of vascular abnormalities and neoplasms]. AB - Vascular malformations and neoplasms are very common skin disorders, found in up to 5% of newborns. However, a clear distinction has to be made between proliferating vascular lesions and permanent malformations. An exact classification is also extremely useful, since many new specific diagnostic and therapeutic measures have been developed in recent years. True proliferating tumors are, for example, childhood hemangiomas, glomus tumors, granuloma pyogenicum, tufted angiomas, senile angiomas, and malignant vascular lesions. Vascular malformations can affect capillaries, veins or arteries, as well as lymphatic vessels. Arteriovenous shunts and combined malformations may also exist (Hamburg classification). Nevi flammei, nevi anaemici, hematolymphangiomas, angiokeratotic nevi, circumscribed venous-arterious malformations, and the blue rubber-blebnevus-syndrome may either be infiltrating or circumscribed and are characterised by a persistence of the primitive vessel network. In contrast, other malformations involve various vascular trunks, showing vessel dilation or obstruction, often combined with changes in bone or soft tissue. Significant large vessel malformations are the Bockenheimer syndrome, the Klippel-Trenaunay syndrome and the Parkes-Weber syndrome. Combinations involving both large trunks and extravascular space such as the Klippel-Trenaunay syndrome also occur. PMID- 9410846 TI - [Skin disease and sexuality. An empirical study of sex behavior or patients with psoriasis vulgaris and neurodermatitis in comparison with skin-healthy probands]. AB - Fifty-three patients with psoriasis, 24 patients with atopic eczema and 52 controls with healthy skin were compared with regard to their sexual behavior. For the test, questionnaires developed by Arentewicz as well as our own questionnaire on sexuality and partnership were used. Patients with skin diseases had a significantly impaired sex life, compared to those with healthy skin. There was a highly significant reduction in the exchange of tenderness in patients of both sexes, and in the capacity for orgasm in female patients. On the other hand, no significant difference was found concerning the frequency of intercourse. Patients with psoriasis felt more impaired than those with atopic eczema. Ninety three percent of psoriatics and 96% of patients with atopic eczema had not been asked about their sexual life by their attending doctor. We discuss to what extent skin diseases are possibly involved in the regulation of nearness and distance among partners. The dermatologist should a be knowledgeable and understanding source for patients which skin diseases who have questions about their disease and its effect on their sex life. PMID- 9410847 TI - [Recurrent thrombophlebitis and ulcera crurum as manifestations of hereditary blood coagulation disorders and Klinefelter syndrome. Discussion based on 4 case examples]. AB - Recurrent phlebothromboses in young patients with subsequent severe postthrombotic syndrome and chronic venous leg ulcers may be caused by underlying hereditary disorders of hemostasis or may occur as part of other congenital syndromes. The most common hereditary disorders of hemostasis in this respect appear to be deficiencies of antithrombin III, protein C, and protein S, and activated protein C resistance (mutations of factor V). Less frequently, dysfibrinogenemia, increased plasminogen activator inhibitor levels, or deficiencies of tissue plasminogen activator or heparin cofactor II may be found. Klinefelter's syndrome and homocystinuria are prime examples of those rare congenital disorders indirectly associated with an elevated risk of thrombosis in young individuals. Early diagnosis of these disorders will allow timely treatment, preventive care and counselling of patients as well as family members. PMID- 9410848 TI - [Bart syndrome--separate entity or a variant of epidermolysis bullosa?]. AB - Bart syndrome was described first by Bart in 1966; it represents the combination of congenital epidermolysis bullosa, congenital localized absence of skin affecting the extremities and shedding or dystrophy of nails. This syndrome may be of clinical relevance because of its more favourable prognosis in comparison with other forms of epidermolysis bullosa. We report two patients with Bart syndrome and focus on the question, if this syndrome represents a distinct entity or a variant of epidermolysis bullosa. PMID- 9410850 TI - [Papular mucinosis--successful therapy with plasmapheresis]. AB - A 23-year old female patient presented with a 14 year history of cutaneous mucinosis. Her hands showed indurated edema with acrocyanosis and severe reduction in motility, while her face was red, edematous and revealed numerous small angiomas. In the sacral region, she had a large elevated skin colored plaque. On the basis of clinical findings, histology and further laboratory tests the disease was classified as papular mucinosis (lichen myxoedematosus). In view of the severe suffering of the young patient and an unsuccessful previous therapy with chloroquine, we decided to employ plasmapheresis as a single therapy. After the first treatment course there was an improvement in her skin condition. After a total of four plasmapheresis courses over a period of 18 months (total exchange volume of 38.4 l) there was an objective flattening of the plaques and reduction of the finger swelling. This case demonstrates that monotherapy of plasmapheresis is an effective mode of treatment for cutaneous mucinosis, a disease which is extremely difficult to treat. PMID- 9410849 TI - [Responders and nonresponders of ultraviolet A-1 therapy of acute exacerbated atopic eczema]. AB - Of 46 patients with severe atopic eczema treated with UVA-1, 14 were evaluated as non-responders and 32 as responders. Responding patients significantly had (p < 0.05) more severe skin involvement before onset of treatment (clinical severity score SCORAD 62.7 +/- 12.4 points) compared to non-responders (53.0 +/- 10.6 points). Serum levels of total IgE, ECP and sIL2R before treatment did not show any significant differences between the two groups. The data confirm the efficacy of UVA-1 treatment in severe cases of atopic eczema. PMID- 9410851 TI - [Combined naevus flammeus and naevus fuscocoeruleus: phacomatosis pigmentovascularis type IIa]. AB - The association of nevus flammeus with mongolian spot, nevus fuscoceruleus, nevus spilus and, with variable frequency, with nevus anemicus has been termed phacomatosis pigmentovascularis, a genodermatosis first described by Ota and co workers. Four different combinations have been specified. Most cases are reported from the Japanese literature. Phacomatosis pigmentovascularis may constitute an exclusively cutaneous disorder, but overlapping with other syndromes like Klippel Trenaunay syndrome or Sturge-Weber syndrome is also possible. We report a 30-year old woman with nevus flammeus on the back and right arm associated with nevus fuscoceruleus on the back. PMID- 9410852 TI - [Metageria--clinical manifestations of a premature aging syndrome]. AB - A 19-year old caucasian patient suffered from ulceration and scaring of his fingers since age two. During childhood, fibrosing contractures on the phalanges developed. Since puberty, his eunuch-like stature, bird-like face and exophthalmos had become under obvious. His hair was extraordinarily fine; his pubic hair rare. Radiological examination revealed ankylosis and osteoporosis of the phalanges and carpals. Angiography showed occlusion of multiple digital and interdigital arteries. Furthermore, a bilateral posterior cataract, restrictive respiratory disease, impaired glucose tolerance, hyperuricemia, proteinuria and primary hypogonadism were diagnosed. These findings are characteristic for the premature aging syndromes. The cardial symptoms of tall stature, bird-like face, pseudoexophthalmos, skleroderma-like and poikiloderma-like cutaneous lesions, scarce hair growth, early diabetes mellitus and arteriosclerosis led to the diagnosis of metageria. PMID- 9410853 TI - [Hypomelanosis guttata idiopathica]. AB - A 37-year-old white female with dark complexion presented with progressive hypopigmented macules on the face and upper trunk. She was in good health and her family history was unremarkable. Histologic examination of the hypopigmented patches revealed reduced melanin within the basal layers of the epidermis. Using immunohistochemical and ultrastructural studies, the number of melanocytes appeared normal but signs of degeneration were evident and dendrites were shortened. The diagnosis of idiopathic guttate hypomelanosis was established. The clinical picture, etiology and treatment of this relatively common but often unrecognized skin disease is discussed. PMID- 9410855 TI - [40 years Dresden Dermatology Clinic--from Academy to University]. PMID- 9410854 TI - [Delayed type allergic reaction to red azo dye in tattooing]. AB - Allergic reactions in tattoos are comparatively rare. In most cases the reactions are caused by different red pigments. While in the past these reactions have been ascribed to mercury salts (cinnebar) and cadmium sulphide, now synthetic inorganic azo dyes have also been found to be responsible for such reactions. A 42-year-old man presented with an allergic reaction in the red parts of his tattoos. Histologically a chronic granulomatous, partly fibrous inflammation with transfollicular elimination of pigment granules was found. Spontaneous regression in a part of the inflammatory reaction was observed, simultaneously with depigmentation and scarring of the overlying skin. The pigment used for tattooing was found to be an aromatic azo derivative. In addition to a positive cutaneous reaction to the dye, the patient also showed a positive patch test to Napthol AS, used for the coupling of different dyes in the textile industry. PMID- 9410856 TI - [Pigmentation disorders in corticoid injections]. PMID- 9410857 TI - [Meeting report. 11th International Symposium on Bioengineering and the Skin 2-5 October 1996, Zurich]. PMID- 9410859 TI - [Dermatology on the Internet]. PMID- 9410858 TI - [Vitiligo]. PMID- 9410860 TI - [Ultrasound in dermatology]. PMID- 9410861 TI - [History of Dermatology. German Society for Control of Venereal Diseases e.V. (DGBG/GBGK 1902)]. PMID- 9410862 TI - Osteopathic manifesto. X. The distinctive difference. 1981. PMID- 9410863 TI - Designer "genes": the future of cattle cloning? PMID- 9410865 TI - Questions and answers regarding veterinarians in disaster response. PMID- 9410864 TI - Assessing pain in animals. PMID- 9410866 TI - Understanding a study of vitamin E administration in dairy cows. PMID- 9410867 TI - Effects of location and information sources on clients' decisions to select or switch veterinary practices. PMID- 9410868 TI - [Radiologic aspects of the loosening of cemented hip prostheses: mechanical, septic or granulomatous etiology?]. AB - Radiologic diagnosis of hip prosthesis loosening is based on the evaluation of each component (prosthesis, cement, bone) and of their interfaces. Both the prosthesis and the cement may deteriorate and the prosthesis/cement interface or cement/bone interface may become abnormal in prosthesis loosening of any etiology. In contrast, the aspect of the bone changes (erosion, periostitis) and their distribution vary according to the condition etiology. It appears from a retrospective study of 50 cases of chronic hip prosthesis loosening that the most specific signs for infection are unsharp bone resorption and acute-like or multifocal periostitis. In granulomatous loosening, bone resorption is sharp (as in mechanical loosening), but its distribution is not conform to the prosthesis shape (as in septic loosening), and periosteal changes are not observed. PMID- 9410869 TI - Suicide assessment and terminology. PMID- 9410870 TI - Proceedings of the 9th meeting on adrenergic mechanisms. Porto, Portugal, 22-25 September 1996. PMID- 9410872 TI - Development, cell differentiation and cancer. Proceedings of a symposium. 9th International Conference. Pisa, Italy, September 28-October 2, 1996. PMID- 9410871 TI - Water-soluble fullerene derivatives attenuate exsanguination-induced bronchoconstriction of guinea-pigs. AB - 1. This study investigated the effects of increased antioxidants (administration of water-soluble fullerenol-1 and pre-exposure to chronic hypoxia) as well as an iron-chelating agent (deferoxamine) on exsanguination-induced noncholinergic airway constriction in guinea-pigs. 2. Fullerenol-1 usually did not cause significant alteration in respiratory function (lung volumes, dynamic respiratory compliance, maximal expiratory flow at 50% total lung capacity (Vmax50), and forced expiratory flow at 0.1 s (FEV 0.1) at low (200 micrograms kg-1) or at high doses (2 mg kg-1), except that it produced a slight bronchial constricting action (decreases in both Vmax 50 and FEV 0.1) at high doses (2 mg kg-1) via intratracheal instillation. 3. Beginning 15 min after exsanguination, there was a marked temporal decrease in FEV 0.1, indicating a gradual increase in airway constriction with time. 4. Administration of either fullerenol-1 or deferoxamine, or pre-exposure to chronic hypoxia significantly ameliorated the exsanguination induced bronchoconstriction. The results provide evidence that oxygen radicals play an important role in exsanguination-induced airway constriction. 5. The significant effects of the increased antioxidants and deferoxamine,however, cannot be explained by the alteration in either tracheal neutral endopeptidase activity or lung tissue substance P level. PMID- 9410873 TI - Live dynamics of Dictyostelium cofilin suggests a role in remodeling actin latticework into bundles. AB - Cofilin, an indispensable, actin-regulating protein represents the 'cofilin family' of actin-binding proteins existing in a wide variety of organisms. Our previous and other in vitro studies have implied that cofilin can accelerate transformation of filamentous (F)-actin and (alpha)-actinin latticework into bundles, and overexpression of cofilin induces formation of F-actin bundles in Dictyostelium. Here we expressed an Aequorea green fluorescent protein (GFP) Dictyostelium cofilin fusion protein in Dictyostelium, and observed the live dynamics to examine the physiological function of cofilin. We show that purified GFP-cofilin binds to actin filaments and decreases the apparent viscosity of actin solution in a similar manner to authentic Dictyostelium cofilin. Expressed GFP-cofilin exhibits normal actin-binding activities in the cytoplasm as represented by incorporation into the actin rods induced with dimethyl sulfoxide. Free moving cells form a crown-like cortical structure on the dorsal surface, and GFP-cofilin exhibits dynamic assembly into actin bundles being formed beneath the cortex. During phagocytosis, GFP-cofilin accumulates into actin bundles formed in the region underlying the phagocytic cups. In cells chemotactically activated with cyclic AMP, GFP-cofilin exhibits a high level of accumulation in projecting leading edges. When the chemo-attraction is experimentally changed, the redistribution of GFP-cofilin towards the new pseudopod occurs in a matter of 30 60 seconds. These results demonstrate that cofilin plays a crucial role in vivo in rapid remodeling of the cortical actin meshwork into bundles. PMID- 9410874 TI - Deregulated expression of the RanBP1 gene alters cell cycle progression in murine fibroblasts. AB - RanBP1 is a molecular partner of the Ran GTPase, which is implicated in the control of several processes, including DNA replication, mitotic entry and exit, cell cycle progression, nuclear structure, protein import and RNA export. While most genes encoding Ran-interacting partners are constitutively active, transcription of the RanBP1 mRNA is repressed in non proliferating cells, is activated at the G1/S transition in cycling cells and peaks during S phase. We report here that forced expression of the RanBP1 gene disrupts the orderly execution of the cell division cycle at several stages, causing inhibition of DNA replication, defective mitotic exit and failure of chromatin decondensation during the telophase-to-interphase transition in cells that achieve nuclear duplication and chromosome segregation. These results suggest that deregulated RanBP1 activity interferes with the Ran GTPase cycle and prevents the functioning of the Ran signalling system during the cell cycle. PMID- 9410875 TI - Roles of plakoglobin end domains in desmosome assembly. AB - Plakoglobin, a member of the armadillo family of proteins, is a component of intercellular adhesive junctions. The central domain of plakoglobin comprises a highly conserved series of armadillo repeats that facilitate its association with either desmosomal or classic cadherins, or with cytosolic proteins such as the tumor suppressor gene product adenomatous polyposis coli. Sequences in the N- and C-terminal domains of plakoglobin are less highly conserved, and their possible roles in regulating plakoglobin's subcellular distribution and junction assembly are still unclear. Here we have examined the role of plakoglobin end domains by stably expressing constructs lacking the N and/or C terminus of plakoglobin in A 431 cells. Our results demonstrate that myc-tagged plakoglobin lacking either end domain is still able to associate with the desmosomal cadherin desmoglein and incorporate into desmosomes. In cell lines that express an N-terminal truncation of plakoglobin, an increase in the cytosolic pool of en-dogenous and ectopic plakoglobin was observed that may reflect an increase in the stability of the protein. Deletion of the N terminus did not have a dramatic effect on the structure of desmosomes in these cells. On the other hand, striking alterations in desmosome morphology were observed in cells expressing C-terminal truncations of plakoglobin. In these cell lines, ectopic plakoglobin incorporated into desmosomes, and extremely long junctions or groups of tandemly linked desmosomes which remained well attached to keratin intermediate filaments, were observed. Together, these results suggest that plakoglobin end domains play a role in regulating its subcellular distribution, and that the presence of the C terminus limits the size of desmosomes, perhaps through regulating protein-protein interactions required for assembly of the desmosomal plaque. PMID- 9410876 TI - Expression of a minus-end-directed motor protein induces Sf9 cells to form axon like processes with uniform microtubule polarity orientation. AB - Neurons extend two types of processes with distinct morphologies and patterns of microtubule polarity orientation. Axons are thin cylindrical processes containing microtubules that are uniformly oriented with their plus-ends-distal to the cell body while dendrites are stout tapering processes that contain nonuniformly oriented microtubules. We have proposed that these distinct microtubule patterns are established by molecular motors that transport microtubules into each type of process with the appropriate orientation. To test the feasibility of this proposal, we have embarked on a series of studies involving the expression of vertebrate motors in insect Sf9 cells. We previously focused on a kinesin-related protein termed CHO1/MKLP1, which localizes to the midzone of the mitotic spindle, and which has been shown to have the appropriate properties to transport microtubules of opposite orientation relative to one another. Expression of a fragment of CHO1/MKLP1 containing its motor domain induces Sf9 cells to extend processes with a stout tapering morphology and a nonuniform microtubule polarity pattern similar to dendrites. Here we focus on a minus-end-directed kinesin related motor protein termed CHO2, which localizes to the non-overlapping regions of the mitotic spindle, and which has been shown to have the appropriate properties to transport microtubules with plus-ends-leading. Sf9 cells induced to express a fragment of CHO2 containing its motor domain extend processes with a long cylindrical morphology and a uniformly plus-end-distal microtubule polarity pattern similar to axons. These results show that motor proteins have the capacity to organize distinct patterns of microtubule polarity orientation during process outgrowth, and that these patterns are intimately related to the unique morphological characteristics of the processes. Moreover, mutation of three amino acids corresponding to the ATP binding site necessary for motor function suppresses the capacity of the CHO2 fragment to induce process formation and microtubule reorganization, indicating that at least in the case of CHO2, the transport properties of the motor are essential for it to elicit these effects. PMID- 9410877 TI - Internalization and recycling of the C5a anaphylatoxin receptor: evidence that the agonist-mediated internalization is modulated by phosphorylation of the C terminal domain. AB - The C5a anaphylatoxin receptor is a member of the G protein-coupled receptor family involved in chemoattraction and activation of myeloid cells, as well as in host defence against infection by Pseudomonas aeruginosa. Upon challenge by C5a, the C5a receptor undergoes a rapid phosphorylation on serine residues in the carboxyl-terminal region. In this study, we used cells stably transfected with either the wild-type C5a receptor, or mutants affected in their capacity to be phosphorylated, to examine the role played by phosphorylation in the intracellular trafficking of the C5a receptor. Upon agonist binding, the wild type receptor was rapidly internalized into endosomes that cluster near the nucleus after 10 minutes. Internalization of a non-phosphorylable mutant was severely impaired relative to wild-type receptor, whereas a mutant phos phorylated on serine 327 and/or serine 338, showed a rate of internalization intermediate between that of wild-type receptor and that of the non phosphorylable mutant. Under continuous exposure to C5a and in the absence of protein synthesis, the C5a receptor was maintained in a highly phosphorylated state but was not degraded. Confocal microscopy and ligand-binding studies indicated that internalized receptors were recycled to the plasma membrane. During this process, receptors were dephosphorylated with kinetics that correlated with the kinetics of receptor recovery on the cell surface. Altogether, our data suggest that phosphorylation plays a key role in the intracellular trafficking of the C5a receptor. Phosphoryl-ated receptors might be recognized by an adaptor protein that interacts with the endocytic machinery. PMID- 9410878 TI - Non-centrosomal microtubule formation and measurement of minus end microtubule dynamics in A498 cells. AB - Experiments performed on a cell line (A498) derived from a human kidney carcinoma revealed non-centrosomal microtubules in the peripheral lamella of many cells. These short microtubules were observed in glutaraldehyde-fixed cells by indirect immunofluorescence, and in live cells injected with rhodamine-labeled tubulin. The non-centrosomal microtubules were observed to form de novo in living cells, and their complete disassembly was also observed. Low-light-level fluorescence microscopy, coupled to imaging software, was utilized to record and measure the dynamic behavior of both ends of the non-centrosomal microtubules in these cells. For each, the plus end was differentiated from the minus end using the ratio of their transition frequencies and by measuring total assembly at each end. For comparative purposes, dynamics of the plus ends of centrosomally nucleated microtubules were also analyzed in this cell line. Our data reveal several striking differences between the plus and minus ends. The average pause duration was nearly 4-fold higher at the minus ends; the percentage of time spent in pause was 92% at the minus ends, compared to 55% at plus ends. Dynamicity was decreased 4-fold at the minus ends, and the average number of events per minute was reduced from 7.0 at the plus end to 1.5 at the minus ends. The minus ends also showed a 6 fold decrease in frequency of catastrophe over the plus ends. These data demonstrate that in living cells, microtubules can form at sites distant from the perinuclear microtubule organizing center, and once formed, non-centrosomal microtubules can persist for relatively long periods. PMID- 9410879 TI - Pulse treatment of interphasic HeLa cells with nanomolar doses of docetaxel affects centrosome organization and leads to catastrophic exit of mitosis. AB - In order to investigate the role of centrosome duplication in mitotic spindle morphogenesis, we designed a 1 hour pulse treatment protocol on synchronized HeLa cells with nanomolar doses of taxoids that might impair centrosome biogenesis but would allow the recovery of normal microtubule (Mt) dynamics before mitosis. We were prompted to use this approach as docetaxel (DOC; taxotereTM), a taxoid known to promote Mt polymerization, was shown to be more cytotoxic when applied during S phase. We show that pulse drug exposure is most efficient in late S and in G2 and results in a marked disorganization of the centrosome in G2, the pericentriolar material (PCM) being dissociated from centrioles. Separation of centrosomes at the G2-M transition is also impaired and mitotic spindle morphogenesis is grossly abnormal: although in most spindles chromosomes align in a metaphase plate, the two centrosomes stay most often unseparated at one pole and most of the NuMA protein accumulates at the other. Interestingly, we find that the centrosomes' ability to duplicate is not abolished as they are still able to trigger parthenogenetic development of frog eggs. Despite spindle asymmetry, the progression through mitosis is not blocked. This results in a catastrophic exit from mitosis, each mitotic cell generating several micronucleated cells linked together by multiple midbodies. Lack of mitotic block appears therefore as the prime cause of cell lethality. These experiments suggest that NuMA redistribution at the onset of mitosis depends upon the correct redistribution of PCM between centriole pairs. They also indicate that the presence of aberrant spindle poles does not alert the surveillance mechanism controlling the exit of mitosis. PMID- 9410880 TI - Dissociation of LPA-induced cytoskeletal contraction from stress fiber formation by differential localization of RhoA. AB - Addition of lysophosphatidic acid (LPA) to serum-deprived N1E-115 neuronal cells results in rapid f-actin assembly accompanied by neurite retraction and rounding of the cell body due to contraction of the cortical actin cytoskeleton. LPA action is mimicked by activated RhoA, while it is blocked by dominant-negative RhoA (N19RhoA) and the Rho-inactivating C3 toxin. Using immunofluorescence analysis and high speed centrifugation we show that activated RhoA is localized to the plasma membrane. Wild-type RhoA and N19RhoA, however, are mainly cytosolic. We find that LPA-induced shape changes are preceded by translocation of RhoA from the cytosol to the cell periphery. LPA also stimulates translocation of inactive N19RhoA in the absence of ensuing shape changes. When membrane localization of RhoA is prevented by lovastatin, an inhibitor of protein isoprenylation, or by CAAX motif mutation, cytoskeletal contraction is blocked. However, the assembly of f-actin into stress fibers is not affected under these conditions. The effects of both LPA and activated RhoA are blocked by tyrosine kinase inhibitors (herbimycin, genistein, tyrphostin), but not by dominant negative Src. We conclude that: (1) LPA-induced cytoskeletal contraction, but not stress fiber formation, requires translocation of RhoA from the cytosol to the plasma membrane; (2) translocation of RhoA occurs independently of its activation; and (3), a non-Src tyrosine kinase is involved in RhoA-stimulated contractility. PMID- 9410881 TI - When rDNA transcription is arrested during mitosis, UBF is still associated with non-condensed rDNA. AB - The mechanisms that control inactivation of ribosomal gene (rDNA) transcription during mitosis is still an open question. To investigate this fundamental question, the precise timing of mitotic arrest was established. In PtK1 cells, rDNA transcription was still active in prophase, stopped in prometaphase until early anaphase, and activated in late anaphase. Because rDNA transcription can still occur in prophase and late anaphase chromosomes, the kinetics of rDNA condensation during mitosis was questioned. The conformation of the rDNA was analyzed by electron microscopy from the G2/M transition to late anaphase in the secondary constriction, the chromosome regions where the rDNAs are clustered. Whether at transcribing or non-transcribing stages, non-condensed rDNA was observed in addition to axial condensed rDNA. Thus, the persistence of this non condensed rDNA during inactive transcription argues in favor of the fact that mitotic inactivation is not the consequence of rDNA condensation. Analysis of the three-dimensional distribution of the rDNA transcription factor, UBF, revealed that it was similar at each stage of mitosis in the secondary constriction. In addition, the colocalization of UBF with non-condensed rDNA was demonstrated. This is the first visual evidence of the association of UBF with non-condensed rDNA. As we previously reported that the rDNA transcription machinery remained assembled during mitosis, the colocalization of rDNA fibers with UBF argues in favor of the association of the transcription machinery with certain rDNA copies even in the absence of transcription. If this hypothesis is correct, it can be assumed that condensation of rDNA as well as dissociation of the transcription machinery from rDNA cannot explain the arrest of rDNA transcription during mitosis. It is proposed that modifications of the transcription machinery occurring in prometaphase could explain the arrest of transcription, while reverse modifications in late anaphase could explain activation. PMID- 9410882 TI - Microtubule depolymerization inhibits clathrin coated-pit internalization in non adherent cell lines while interleukin 2 endocytosis is not affected. AB - The microtubule cytoskeleton is generally not considered to be essential for the first steps of clathrin-mediated endocytosis of membrane receptors. Its role in clathrin-independent endocytosis has not been investigated. We have previously shown that the cytokine interleukin 2 (IL2) is internalized in lymphoid cells expressing its receptors when clathrin-dependent endocytosis is inhibited. Here we compare the internalization of IL2 and of transferrin, a marker of clathrin dependent endocytosis, after microtubule disruption. In hemopoietic cell lines, which express IL2 receptors, transferrin receptor entry was inhibited by about 40%. However, in adherent cell lines, transferrin entry was unaffected by microtubule disruption, as previously reported. Unlike the case for transferrin, internalization of IL2 receptors was not affected by depolymerization of the microtubule cytoskeleton in hemopoietic cell lines. These results show that IL2 and transferrin receptors do not have the same endocytic properties and support our previous conclusion that these receptors follow different pathways of endocytosis. PMID- 9410883 TI - Ultrastructural localization of cPLA2 in unstimulated and EGF/A23187-stimulated fibroblasts. AB - The 85 kDa cytosolic phospholipase A2 is the key enzyme in the release of arachidonic acid. To gain insight into cytosolic phospholipase A2 action in mitogen-activated cells, the localization of the phospholipase was investigated in fibroblasts upon stimulation with epidermal growth factor and the calcium ionophore A23187. By the use of indirect immunofluorescence microscopy, staining of endogenous cytosolic phospholipase A2 resulted in a punctate labeling pattern randomly distributed throughout the cytoplasm of the cell. Immunogold electron microscopy revealed that this punctate labeling pattern exhibited the presence of the 85 kDa phospholipase A2 in small clusters. These clusters were found in the cytosol in the vicinity of all organellar membranes, except for the Golgi system. The enzyme showed no preference for the nuclear envelope, the endoplasmic reticulum or the plasma membrane. Stimulation of cells with epidermal growth factor or A23187 or both did not change the punctate immunofluorescence labeling pattern. Furthermore, a similar labeling pattern was observed by the artificial introduction of extremely low or high intracellular calcium concentrations. Even by electron microscopy, translocation of cytosolic phospholipase A2 to membranes was not observed after stimulation of cells with epidermal growth factor and A23187. From these results it is concluded that cytosolic phospholipase A2 is localized in clusters close to membranes in stimulated as well as unstimulated fibroblasts, without preference for a specific organellar membrane. PMID- 9410884 TI - Reduced expression of tissue transglutaminase in a human endothelial cell line leads to changes in cell spreading, cell adhesion and reduced polymerisation of fibronectin. AB - Tissue transglutaminase (tTgase, type II) is a Ca2+-dependent GTP binding protein which crosslinks proteins via (epsilon)((gamma)-glutamyl)lysine bridges. Although essentially a cytosolic enzyme there is increasing evidence to suggest the enzyme is externalised where it may play a role in extracellular matrix organisation. To investigate the function of this enzyme in a human umbilical endothelial cell line ECV304 tTgase expression was reduced in these cells by up to 90% by stable transfection with a 1.1. kb antisense construct in the plasmid vector pSG5. Two clones showing a reduction in expression of tTgase activity of 70 and 90% have been isolated and characterised. These clones show a number of phenotypic differences when compared to the parent cell line and the transfected controls which include reduced cell spreading and a decreased adhesion of cells on different substrata as measured by their susceptibility to removal by trypsin. Reduced cell spreading in the antisense transfected clones was accompanied by a decrease in the crosslinking of fibronectin into polymeric multimers which could be correlated to the amount of tTgase externalised by cells. A novel assay was developed to measure externalised tTgase activity which is cell mediated, inhibited by preincubation of cells with anti-tTgase antibody and relies on the incorporation of biotinylated cadaverine into fibronectin. The results of these experiments suggest that externalised tTgase may play a key role in a number of cell behavioural patterns which might be related to the enzymes ability to bind and crosslink fibronectin. PMID- 9410885 TI - Matrix metalloproteinase-2 activation modulates glioma cell migration. AB - Stable transfection of U251.3 glioma cells with cDNA encoding MT-MMP-1 resulted in increased cell surface expression of MT-MMP-1 and TIMP-2, constitutive activation of MMP-2 proenzyme and increased collagen degradation. In tumor spheroid outgrowth assays, cell migration of MT-MMP-1 transfectants relative to control was enhanced on collagen and decreased on vitronectin and fibronectin. These effects were reversed by TIMP-2 and were not associated with any substantial changes in cell adhesion. Binding of U251.3 cells to the C-terminal domain of MMP-2 was specifically inhibited by anti-(alpha)vss3 integrin blocking antibody indicating that MMP-2 interacts with (alpha)vss3 through the enzyme's C terminal portion at or near the integrin's matrix adhesion sites. We propose that these mechanisms could govern directed matrix degradation in the tumor cells' microenvironment by sequestration of active MMP-2 on the cell surface. Our data suggest that activation of MMP-2 and its proteolytic activity localized to the cell surface could differentially modulate tumor cell migration in response to particular matrix proteins by altering both composition of the extracellular matrix and expression of adhesion receptors on the cell surface. PMID- 9410887 TI - Dispersal of Golgi apparatus in nocodazole-treated fibroblasts is a kinesin driven process. AB - The morphology and location of the Golgi apparatus (GA) has been shown to change upon microtubule (Mt) depolymerization. The GA in different cell types undergoes fragmentation and dispersal throughout the cytoplasm upon treatment with nocodazole. In this study experiments were performed on human skin fibroblasts (HSFs) and rat fibroblasts (REF 52) to determine whether the dispersal of GA in HSFs treated with nocodazole is dependent on Mts that show the higher resistance to this Mt-depolymerizing drug. It is shown here that nocodazole at concentrations as low as 100 nM caused the GA to disperse in treated fibro-blasts that still contained a fairly high amount of Mts. Antibody-blocking analysis of Mts after injection of biotin-tubulin into the HSFs was used to show that nocodazole at low concentrations induced the stabilization of the remaining Mts. The complete disruption of Mts by the incubation of HSFs at 0 degrees C prevented the dispersal of GA from the perinuclear area when the cells were subsequently warmed to 37 degrees C in the presence of nocodazole. Micro-injection of the well characterized HD antibody against kinesin but not the preimmune IgG caused inhibition of GA dispersal in HSFs by nocodazole. These data demonstrate that the dispersal of GA in the cytoplasm of nocodazole-treated HSFs is a kinesin-driven process with stable Mts serving as tracks. PMID- 9410888 TI - Medical contradictions: or, what's a body to do? PMID- 9410886 TI - Distinct nuclear assembly pathways for lamins A and C lead to their increase during quiescence in Swiss 3T3 cells. AB - The expression of A-type lamins coincides with cell differentiation and as A-type lamins specifically interact with chromatin, a role in the regulation of differential gene expression has been suggested for A-type lamins. Using the mouse Swiss 3T3 cell line as a model, the change in two A-type lamins, lamins A and C, during cellular quiescence has been investigated. This well established model system mimics the first stages of differentiation when cells exit the cell cycle. In fact, quiescence in Swiss 3T3 cells was accompanied by a significant increase (2.6-fold) in lamin A protein levels and a smaller but reproducible increase (1.4-fold) in lamin C. These effects were fully reversible upon restimulation of the cells with serum. No effect upon lamin B levels was observed. Conversely, levels of A-type lamin mRNA decreased markedly as a result of quiescence suggesting transcriptional mechanisms are involved in the change in levels of lamins A and C. No difference in the incorporation of microinjected human lamin A into nuclei of quiescent or proliferating cells was observed. These data suggest A-type lamin binding sites were not limiting and indicated little difference between A-type lamin assembly mechanisms in quiescent and proliferating cells. The data did demonstrate lamin A and lamin C incorporation into the nuclear lamina proceeded by different pathways when microinjected in Swiss 3T3 cells. The incorporation of recombinant lamin C into the nuclear lamina was delayed compared to lamin A and proceeded via intranuclear foci. Such foci were not seen with microinjected lamin A. Instead, recombinant lamin A was rapidly (<20 minutes) incorporated into the nuclear lamina. Comicroinjection of lamin A with lamin C did not prevent foci formation but assisted in the rapid clearing (t1/2=30 minutes) of these structures and the incorporation of both lamins A and C into the lamina. These data suggest that the incorporation of lamin C into the lamina is facilitated by lamin A. They demonstrate a distinct difference in the nuclear assembly pathways of lamins A and C and show for the first time a functional distinction for these two splice variants of the A-type lamin gene. From the differences in assembly pathways and changes in protein levels accompanying quiescence in 3T3 cells, we suggest distinct roles for lamin A and lamin C in proliferating and quiescent states of the cell cycle. PMID- 9410889 TI - Atherosclerosis, just another cancer? PMID- 9410890 TI - Nitric oxide synthases: which, where, how, and why? PMID- 9410891 TI - Cellular and molecular mechanisms of endothelial cell dysfunction. PMID- 9410893 TI - The atherogenic lipoprotein Lp(a) is internalized and degraded in a process mediated by the VLDL receptor. AB - Lp(a) is a major inherited risk factor associated with premature heart disease and stroke. The mechanism of Lp(a) atherogenicity has not been elucidated, but likely involves both its ability to influence plasminogen activation as well as its atherogenic potential as a lipoprotein particle after receptor-mediated uptake. We demonstrate that fibroblasts expressing the human VLDL receptor can mediate endocytosis of Lp(a), leading to its degradation within lysosomes. In contrast, fibroblasts deficient in this receptor are not effective in catabolizing Lp(a). Lp(a) degradation was prevented by antibodies against the VLDL receptor, and by RAP, an antagonist of ligand binding to the VLDL receptor. Catabolism of Lp(a) was inhibited by apolipoprotein(a), but not by LDL or by monoclonal antibodies against apoB100 that block LDL binding to the LDL receptor, indicating that apolipoprotein(a) mediates Lp(a) binding to this receptor. Removal of Lp(a) antigen from the mouse circulation was delayed in mice deficient in the VLDL receptor when compared with control mice, indicating that the VLDL receptor may play an important role in Lp(a) catabolism in vivo. We also demonstrate the expression of the VLDL receptor in macrophages present in human atherosclerotic lesions. The ability of the VLDL receptor to mediate endocytosis of Lp(a) could lead to cellular accumulation of lipid within macrophages, and may represent a molecular basis for the atherogenic effects of Lp(a). PMID- 9410894 TI - Genomic instability in the type II TGF-beta1 receptor gene in atherosclerotic and restenotic vascular cells. AB - Cells proliferating from human atherosclerotic lesions are resistant to the antiproliferative effect of TGF-beta1, a key factor in wound repair. DNA from human atherosclerotic and restenotic lesions was used to test the hypothesis that microsatellite instability leads to specific loss of the Type II receptor for TGF beta1 (TbetaR-II), causing acquired resistance to TGF-beta1. High fidelity PCR and restriction analysis was adapted to analyze deletions in an A10 microsatellite within TbetaR-II. DNA from lesions, and cells grown from lesions, showed acquired 1 and 2 bp deletions in TbetaR-II, while microsatellites in the hMSH3 and hMSH6 genes, and hypermutable regions of p53 were unaffected. Sequencing confirmed that these deletions occurred principally in the replication error-prone A10 microsatellite region, though nonmicrosatellite mutations were observed. The mutations could be identified within specific patches of the lesion, while the surrounding tissue, or unaffected arteries, exhibited the wild type genotype. This microsatellite deletion causes frameshift loss of receptor function, and thus, resistance to the antiproliferative and apoptotic effects of TGF-beta1. We propose that microsatellite instability in TbetaR-II disables growth inhibitory pathways, allowing monoclonal selection of a disease-prone cell type within some vascular lesions. PMID- 9410892 TI - Angiotensin II inhibits insulin signaling in aortic smooth muscle cells at multiple levels. A potential role for serine phosphorylation in insulin/angiotensin II crosstalk. AB - To investigate potential interactions between angiotensin II (AII) and the insulin signaling system in the vasculature, insulin and AII regulation of insulin receptor substrate-1 (IRS-1) phosphorylation and phosphatidylinositol (PI) 3-kinase activation were examined in rat aortic smooth muscle cells. Pretreatment of cells with AII inhibited insulin-stimulated PI 3-kinase activity associated with IRS-1 by 60%. While AII did not impair insulin-stimulated tyrosine phosphorylation of the insulin receptor (IR) beta-subunit, it decreased insulin-stimulated tyrosine phosphorylation of IRS-1 by 50%. AII inhibited the insulin-stimulated association between IRS-1 and the p85 subunit of PI 3-kinase by 30-50% in a dose-dependent manner. This inhibitory effect of AII on IRS-1/PI 3 kinase association was blocked by the AII receptor antagonist saralasin, but not by AT1 antagonist losartan or AT2 antagonist PD123319. AII increased the serine phosphorylation of both the IR beta-subunit and IRS-1. In vitro binding experiments showed that autophosphorylation increased IR binding to IRS-1 from control cells by 2.5-fold versus 1.2-fold for IRS-1 from AII-stimulated cells, suggesting that AII stimulation reduces IRS-1's ability to associate with activated IR. In addition, AII increased p85 serine phosphorylation, inhibited the total pool of p85 associated PI 3-kinase activity, and decreased levels of the p50/p55 regulatory subunit of PI 3-kinase. These results suggest that activation of the renin-angiotensin system may lead to insulin resistance in the vasculature. PMID- 9410895 TI - Expression of protein kinase C beta in the heart causes hypertrophy in adult mice and sudden death in neonates. AB - Protein kinase C (PKC) activation in the heart has been linked to a hypertrophic phenotype and to processes that influence contractile function. To establish whether PKC activation is sufficient to induce an abnormal phenotype, PKCbeta was conditionally expressed in cardiomyocytes of transgenic mice. Transgene expression in adults caused mild and progressive ventricular hypertrophy associated with impaired diastolic relaxation, whereas expression in newborns caused sudden death associated with marked abnormalities in the regulation of intracellular calcium. Thus, the PKC signaling pathway in cardiocytes has different effects depending on the timing of expression and, in the adult, is sufficient to induce pathologic hypertrophy. PMID- 9410896 TI - Abnormal sulfate metabolism in vitamin D-deficient rats. AB - To explore the possibility that vitamin D status regulates sulfate homeostasis, plasma sulfate levels, renal sulfate excretion, and the expression of the renal Na-SO4 cotransporter were evaluated in vitamin D-deficient (D-D-) rats and in D-D rats rendered normocalcemic by either vitamin D or calcium/lactose supplementation. D-D- rats had significantly lower plasma sulfate levels than control animals (0.93+/-0.01 and 1.15+/-0.05 mM, respectively, P < 0.05), and fractional sulfate renal excretion was approximately threefold higher comparing D D- and control rats. A decrease in renal cortical brush border membrane Na-SO4 cotransport activity, associated with a parallel decrease in both renal Na-SO4 cotransport protein and mRNA content (78+/-3 and 73+/-3% decreases, respectively, compared with control values), was also observed in D-D- rats. Vitamin D supplementation resulted in a return to normal of plasma sulfate, fractional sulfate excretion, and both renal Na-SO4 cotransport mRNA and protein. In contrast, renal sulfate excretion and renal Na-SO4 cotransport activity, protein abundance, and mRNA remained decreased in vitamin D-depleted rats fed a diet supplemented with lactose and calcium, despite that these rats were normocalcemic, and had significantly lower levels of parathyroid hormone and 25(OH)- and 1,25(OH)2-vitamin D levels than the vitamin D-supplemented groups. These results demonstrate that vitamin D modulates renal Na-SO4 sulfate cotransport and sulfate homeostasis. The ability of vitamin D status to regulate Na-SO4 cotransport appears to be a direct effect, and is not mediated by the effects of vitamin D on plasma calcium or parathyroid hormone levels. Because sulfate is required for synthesis of essential matrix components, abnormal sulfate metabolism in vitamin D-deficient animals may contribute to producing some of the abnormalities observed in rickets and osteomalacia. PMID- 9410897 TI - Disruption of the splicing enhancer sequence within exon 27 of the dystrophin gene by a nonsense mutation induces partial skipping of the exon and is responsible for Becker muscular dystrophy. AB - The mechanism of exon skipping induced by nonsense mutations has not been well elucidated. We now report results of in vitro splicing studies which disclosed that a particular example of exon skipping is due to disruption of a splicing enhancer sequence located within the exon. A nonsense mutation (E1211X) due to a G to T transversion at the 28th nucleotide of exon 27 (G3839T) was identified in the dystrophin gene of a Japanese Becker muscular dystrophy case. Partial skipping of the exon resulted in the production of truncated dystrophin mRNA, although the consensus sequences for splicing at both ends of exon 27 were unaltered. To determine how E1211X induced exon 27 skipping, the splicing enhancer activity of purine-rich region within exon 27 was examined in an in vitro splicing system using chimeric doublesex gene pre-mRNA. The mutant sequence containing G3839T abolished splicing enhancer activity of the wild-type purine rich sequence for the upstream intron in this chimeric pre-mRNA. An artificial polypurine oligonucleotide mimicking the purine-rich sequence of exon 27 also showed enhancer activity that was suppressed by the introduction of a T nucleotide. Furthermore, the splicing enhancer activity was more markedly inhibited when a nonsense codon was created by the inserted T residue. This is the first evidence that partial skipping of an exon harboring a nonsense mutation is due to disruption of a splicing enhancer sequence. PMID- 9410898 TI - Human pulmonary alveolar proteinosis associated with a defect in GM-CSF/IL-3/IL-5 receptor common beta chain expression. AB - Pulmonary alveolar proteinosis (PAP) is a heterogeneous disorder of genetic or acquired etiologies. In some cases congenital PAP is associated with hereditary surfactant protein (SP)-B deficiency. To date, the molecular defect in the majority of patients with PAP has not been identified. In mice, PAP has been generated by targeted deletion of the genes for either the GM-CSF/IL-3/IL-5 receptor common beta chain (beta c) or GM-CSF. Here, we describe an expression defect of beta c in three of seven pediatric patients with PAP and in one patient with severe lung disease suspected to be PAP. The patients failed to express normal levels of beta c as shown by flow cytometry. Strikingly reduced or absent function of beta c was demonstrated by ligand binding studies and progenitor clonogenic assays. Analysis of beta c DNA revealed a point mutation from proline to threonine at codon 602 in one patient. Our findings provide evidence that a defect in the expression of a hematopoietic cytokine receptor is associated with human PAP. PMID- 9410899 TI - Phase I trial of recombinant adenovirus gene transfer in lung cancer. Longitudinal study of the immune responses to transgene and viral products. AB - Animal studies indicate that the use of replication-deficient adenovirus for human gene therapy is limited by host antivector immune responses that result in transient recombinant protein expression and blocking of gene transfer when rechallenged. Therefore, we have examined immune responses to an adenoviral vector and to the beta-galactosidase protein in four patients with lung cancer given a single intratumor injection of 10(9) plaque-forming units of recombinant adenovirus. The beta-galactosidase protein was expressed in day-8 tumor biopsies from all patients at variable levels. Recombinant virus DNA was detected by PCR in day-30 and day-60 tumor biopsies from all patients except patient 1. A high level of neutralizing antiadenovirus antibodies was detected in patient 1 before Ad-beta-gal injection whereas it was low (patient 3) or undetectable in the other two patients. All patients developed potent CD4 type 1 helper T cell (Th1) responses to adenoviral particles which increased gradually over time after injection. Antiadenovirus cytotoxic T lymphocyte responses were consistently boosted in the two patients examined (patients 3 and 4). Sustained production of anti-beta-galactosidase IgG was observed in all patients except patient 1. Consistent with anti-beta-gal antibody production, all patients except patient 1 developed intense, dose-dependent Th1 responses to soluble beta-galactosidase which increased over time. Strong beta-galactosidase-specific cytotoxic T lymphocyte responses were detected in patients 2, 3, and 4. Our results clearly show that despite the intensity of antiadenovirus responses, transgene protein expression was sufficient to induce strong and prolonged immunity in three patients. Recombinant adenovirus injected directly into the tumor is a highly efficient vector for immunizing patients against the transgene protein. PMID- 9410900 TI - HLA-DQB1 polymorphism determines incidence, onset, and severity of collagen induced arthritis in transgenic mice. Implications in human rheumatoid arthritis. AB - Certain HLA-DR alleles have been associated with predisposition to human rheumatoid arthritis (RA). There is also evidence that certain HLA-DQ alleles may also be important in determining susceptibility to RA. We have previously demonstrated that mice transgenic for HLA-DQ8, a DQ allele associated with susceptibility to RA, develop severe arthritis after type II collagen immunization. To investigate the influence of polymorphic difference at the DQ loci on susceptibility to arthritis, we generated mice transgenic for HLA-DQ6, an allele associated with a nonsusceptible haplotype. The DQ6 mice were found to be resistant to collagen-induced arthritis. We also assessed the combined effect of an RA-susceptible and an RA nonassociated DQ allele by producing double transgenic mice expressing DQ6 and DQ8 molecules, representing the more prevalent condition found in humans where heterozygosity at the DQ allele is common. The double-transgenic mice developed moderate CIA when immunized with CII when compared with the severe arthritis observed in DQ8 transgenic mice, much like RA patients bearing both susceptible and nonsusceptible HLA haplotypes. These studies support a role for HLA-DQ polymorphism in human RA. PMID- 9410901 TI - Macula densa arginine delivery and uptake in the rat regulates glomerular capillary pressure. Effects of salt intake. AB - These studies tested the hypothesis that delivery and/or cellular uptake of L arginine limits macula densa nitric oxide generation and actions on tubuloglomerular feedback (TGF) during salt restriction. Maximal TGF responses were assessed from reductions in proximal stop flow pressure during loop of Henle (LH) perfusion at 40 nl/min with artificial tubular fluid containing vehicles or drugs. Orthograde LH perfusion of L-arginine (10[-3] M) reduced maximal TGF significantly in rats adapted to low salt (LS: 7.9+/-0.4-6.3+/-0.4 mmHg; P < 0.05), but not high salt (HS: 5.8+/-0.3-5.9+/-0.3; NS). The effects were stereospecific and prevented by coperfusion with NG-methyl-L-arginine. Microperfusion of L-arginine (10[-3] M) into the peritubular capillaries reduced the maximum TGF response more in nephrons of LS than HS rats (deltaTGF: LS, 32+/ 6 vs. HS, 13+/-4%; P < 0.05) and restored a TGF response to luminal perfusion of NG-methyl-L-arginine in LS rats. Coperfusion of nephrons with excess L-lysine or L-homoarginine, which compete with L-arginine for system y+ transport, blocked the fall in proximal stopflow pressure produced by orthograde LH perfusion of L arginine in LS rats. Reabsorption of [3H]arginine by the perfused loop segment was similar in LS (93+/-2%) and HS (94+/-1%) rats. Coperfusion with excess L arginine, L-lysine, or L-homoarginine, however, reduced [3H]arginine reabsorption significantly (P < 0.05) more in HS rats than in LS rats. In conclusion, blunting of maximal TGF responses in salt-restricted rats by nephron-derived NO is limited by L-arginine availability and cellular uptake via system y+. PMID- 9410902 TI - Neonatal activation of CD28 signaling overcomes T cell anergy and prevents autoimmune diabetes by an IL-4-dependent mechanism. AB - Optimal T cell responsiveness requires signaling through the T cell receptor (TCR) and CD28 costimulatory receptors. Previously, we showed that T cells from autoimmune nonobese diabetic (NOD) mice display proliferative hyporesponsiveness to TCR stimulation, which may be causal to the development of insulin-dependent diabetes mellitus (IDDM). Here, we demonstrate that anti-CD28 mAb stimulation restores complete NOD T cell proliferative responsiveness by augmentation of IL-4 production. Whereas neonatal treatment of NOD mice with anti-CD28 beginning at 2 wk of age inhibits destructive insulitis and protects against IDDM by enhancement of IL-4 production by islet-infiltrating T cells, administration of anti-CD28 beginning at 5-6 wk of age does not prevent IDDM. Simultaneous anti-IL-4 treatment abrogates the preventative effect of anti-CD28 treatment. Thus, neonatal CD28 costimulation during 2-4 wk of age is required to prevent IDDM, and is mediated by the generation of a Th2 cell-enriched nondestructive environment in the pancreatic islets of treated NOD mice. Our data support the hypothesis that a CD28 signal is requisite for activation of IL-4-producing cells and protection from IDDM. PMID- 9410903 TI - Pretreatment with antibody to eosinophil major basic protein prevents hyperresponsiveness by protecting neuronal M2 muscarinic receptors in antigen challenged guinea pigs. AB - In antigen-challenged guinea pigs there is recruitment of eosinophils into the lungs and to airway nerves, decreased function of inhibitory M2 muscarinic autoreceptors on parasympathetic nerves in the lungs, and airway hyperresponsiveness. A rabbit antibody to guinea pig eosinophil major basic protein was used to determine whether M2 muscarinic receptor dysfunction, and the subsequent hyperresponsiveness, are due to antagonism of the M2 receptor by eosinophil major basic protein. Guinea pigs were sensitized, challenged with ovalbumin and hyperresponsiveness, and M2 receptor function tested 24 h later with the muscarinic agonist pilocarpine. Antigen-challenged guinea pigs were hyperresponsive to electrical stimulation of the vagus nerves compared with controls. Likewise, loss of M2 receptor function was demonstrated since the agonist pilocarpine inhibited vagally-induced bronchoconstriction in control but not challenged animals. Pretreatment with rabbit antibody to guinea pig eosinophil major basic protein prevented hyperresponsiveness, and protected M2 receptor function in the antigen-challenged animals without inhibiting eosinophil accumulation in the lungs or around the nerves. Thus, hyperresponsiveness is a result of inhibition of neuronal M2 muscarinic receptor function by eosinophil major basic protein in antigen-challenged guinea pigs. PMID- 9410905 TI - Tissue factor expression in human arterial smooth muscle cells. TF is present in three cellular pools after growth factor stimulation. AB - Tissue factor (TF) is a transmembrane glycoprotein that initiates the coagulation cascade. Because of the potential role of TF in mediating arterial thrombosis, we have examined its expression in human aortic and coronary artery smooth muscle cells (SMC). TF mRNA and protein were induced in SMC by a variety of growth agonists. Exposure to PDGF AA or BB for 30 min provided all of the necessary signals for induction of TF mRNA and protein. This result was consistent with nuclear runoff analyses, demonstrating that PDGF-induced TF transcription occurred within 30 min. A newly developed assay involving binding of digoxigenin labeled FVIIa (DigVIIa) and digoxigenin-labeled Factor X (DigX) was used to localize cellular TF. By light and confocal microscopy, prominent TF staining was seen in the perinuclear cytoplasm beginning 2 h after agonist treatment and persisting for 10-12 h. Surface TF activity, measured on SMC monolayers under flow conditions, increased transiently, peaking 4-6 h after agonist stimulation and returning to baseline within 16 h. Peak surface TF activity was only approximately 20% of total TF activity measured in cell lysates. Surface TF blocking experiments demonstrated that the remaining TF was found as encrypted surface TF, and also in an intracellular pool. The relatively short-lived surface expression of TF may be critical for limiting the thrombotic potential of intact SMC exposed to growth factor stimulation. In contrast, the encrypted surface and intracellular pools may provide a rich source of TF under conditions associated with SMC damage, such as during atherosclerotic plaque rupture or balloon arterial injury. PMID- 9410904 TI - Susceptibility to anti-glomerular basement membrane disease and Goodpasture syndrome is linked to MHC class II genes and the emergence of T cell-mediated immunity in mice. AB - We developed a new mouse model of human anti-glomerular basement membrane (GBM) disease to better characterize the genetic determinants of cell-mediated injury. While all major histocompatibility complex (MHC) haplotypes (H-2a, k, s, b, and d) immunized with alpha3 NC1 domains of type IV collagen produce anti-alpha3(IV) NC1 antibodies that cross-react with human Goodpasture [anti-GBM/anti-alpha3(IV) NC1] autoantibodies, only a few strains developed nephritis and lung hemorrhage associated with Goodpasture syndrome. Crescentic glomerulonephritis and lung hemorrhage were MHC-restricted in haplotypes H-2s, b, and d (A beta/A alpha region in H-2s) and associated with the emergence of an IL-12/Th1-like T cell phenotype. Lymphocytes or anti-alpha3(IV) NC1 antibodies from nephritogenic strains transfer disease to syngeneic recipients. However, passive transfer of isogenic alpha3(IV) NC1 antibodies into -/- T cell receptor-deficient mice failed to produce nephritis. Finally, nephritis and its associated IL-12/Th1-like T cell response attenuate in disease-susceptible mice tolerized orally to alpha3(IV) collagen before immunization. Our findings suggest collectively, as a hypothesis, that anti-GBM antibodies in mice only facilitate disease in MHC haplotypes capable of generating nephritogenic lymphocytes with special T cell repertoires. PMID- 9410906 TI - Transgenic expression of the human amphiregulin gene induces a psoriasis-like phenotype. AB - Amphiregulin (AR) is a heparin-binding, heparin-inhibited member of the epidermal growth factor (EGF) family and an autocrine growth factor for human keratinocytes. Previous studies have shown that AR expression is increased in psoriatic epidermis. To test the hypothesis that aberrant AR expression is central to the development of psoriatic lesions, we constructed a transgene (K14 ARGE) encoding a human keratin 14 promoter-driven AR gene. Our results indicate that transgene integration and subsequent expression of AR in basal keratinocytes correlated with a psoriasis-like skin phenotype. Afflicted mice demonstrated shortened life spans, prominent scaling and erythematous skin with alopecia, and occasional papillomatous epidermal growths. Histologic examination revealed extensive areas of marked hyperkeratosis with focal parakeratosis, acanthosis, dermal and epidermal lymphocytic and neutrophilic infiltration, and dilated blood vessels within the papillary dermis. Our results reveal that AR exerts activity in the skin that is distinct from that of transgenic transforming growth factor alpha or other cytokines, and induces skin pathology with striking similarities to psoriasis. Our observations also link the keratinocyte EGF receptor-ligand system to psoriatic inflammation, and suggest that aberrant expression of AR in the epidermis may represent a critical step in the development or propagation of psoriatic lesions. PMID- 9410907 TI - Endogenous somatostatin-28 modulates postprandial insulin secretion. Immunoneutralization studies in baboons. AB - Somatostatin-28 (S-28), secreted into the circulation from enterocytes after food, and S-14, released mainly from gastric and pancreatic D cells and enteric neurons, inhibit peripheral cellular functions. We hypothesized that S-28 is a humoral regulator of pancreatic B cell function during nutrient absorption. Consistent with this postulate, we observed in baboons a two to threefold increase in portal and peripheral levels of S-28 after meals, with minimal changes in S-14. We attempted to demonstrate a hormonal effect of these peptides by measuring their concentrations before and after infusing a somatostatin specific monoclonal antibody (mAb) into baboons and comparing glucose, insulin, and glucagon-like peptide-1 levels before and for 4 h after intragastric nutrients during a control study and on 2 d after mAb administration (days 1 and 2). Basal growth hormone (GH) and glucagon levels and parameters of insulin and glucose kinetics were also measured. During immunoneutralization, we found that (a) postprandial insulin levels were elevated on days 1 and 2; (b) GH levels rose immediately and were sustained for 28 h, while glucagon fell; (c) basal insulin levels were unchanged on day 1 but were increased two to threefold on day 2, coincident with decreased insulin sensitivity; and (d) plasma glucose concentrations were similar to control values. We attribute the eventual rise in fasting levels of insulin to its enhanced secretion in compensation for the heightened insulin resistance from increased GH action. Based on the elevated postmeal insulin levels after mAb administration, we conclude that S-28 participates in the enteroinsular axis as a decretin to regulate postprandial insulin secretion. PMID- 9410908 TI - Pathogenic antibodies inhibit the binding of apolipoproteins to megalin/gp330 in passive Heymann nephritis. AB - Megalin/gp330 is an endocytic receptor that internalizes multiple ligands including apolipoproteins E (apo E) and B100 (apo B). Megalin is the main antigenic target in passive Heymann nephritis (pHN), where it binds circulating autoantibodies leading to the formation of subepithelial immune deposits (ID)-the hallmark of pHN. Apo E and apo B were found recently to accumulate within these IDs, and evidence was provided that their lipids may undergo peroxidation, causing glomerular basement membrane damage and proteinuria. Here we investigated if ID-forming antimegalin IgG can inhibit the binding and internalization of apo E-betaVLDL (very low density lipoprotein) by megalin, and lead to their accumulation within IDs. By immunoelectron microscopy, apo E and apo B were detected in clathrin-coated pits and multivesicular bodies of podocytes in control rats, suggesting that the uptake of lipoproteins is a constitutive function of the glomerular epithelium. When pHN was induced by intravenous injection of antimegalin IgG, apo E and apo B were found within IDs by immunofluorescence and immunoelectron microscopy. Bound antibodies eluted from glomeruli of rats with pHN were found to inhibit the binding and internalization of apo E-enriched betaVLDL by megalin. These results indicate that pHN-inducing antimegalin IgG is capable of interfering with the uptake of lipoproteins by megalin in vivo during the formation of IDs. PMID- 9410909 TI - Angiotensin II stimulates proliferation of normal early erythroid progenitors. AB - Angiotensin II exerts a mitogenic effect in several in vitro models, but a direct effect on erythroid progenitors has not been documented. Angiotensin-converting enzyme inhibitors and losartan, an angiotensin II type 1 receptor (AT1) antagonist, ameliorate posttransplant erythrocytosis, without altering serum erythropoietin levels. We studied erythroid differentiation and the effect of angiotensin II on proliferation of erythroid progenitors by culturing CD34+ hematopoietic progenitor cells in liquid serum-free medium favoring growth of erythroid precursors. Aliquots of cells were collected every third day, and were used for RNA preparation. AT1 mRNA was detected after 6 d. In these same samples, erythroid-specific mRNA (erythropoietin receptor) was also detected. AT1 protein was detected in 7-d-old burst-forming units-erythroid colonies by Western blotting. The CD34+ cell liquid cultures were used to incubate erythroid precursors with angiotensin II from days 6-9. After incubation, cells were transferred to semisolid medium and cultured with erythropoietin. Angiotensin II increased proliferation of early erythroid progenitors, defined as increased numbers of burst-forming units-erythroid colonies. Losartan completely abolished this stimulatory effect of angiotensin II. Moreover, we observed increased numbers of erythroid progenitors in the peripheral blood of posttransplant erythrocytosis patients. Thus, activation of AT1 with angiotensin II enhances erythropoietin-stimulated erythroid proliferation in vitro. A putative defect in the angiotensin II/AT1 pathway may contribute to the pathogenesis of posttransplant erythrocytosis. PMID- 9410910 TI - Changes in gene expression in the intact human heart. Downregulation of alpha myosin heavy chain in hypertrophied, failing ventricular myocardium. AB - Using quantitative RT-PCR in RNA from right ventricular (RV) endomyocardial biopsies from intact nonfailing hearts, and subjects with moderate RV failure from primary pulmonary hypertension (PPH) or idiopathic dilated cardiomyopathy (IDC), we measured expression of genes involved in regulation of contractility or hypertrophy. Gene expression was also assessed in LV (left ventricular) and RV free wall and RV endomyocardium of hearts from end-stage IDC subjects undergoing heart transplantation or from nonfailing donors. In intact failing hearts, downregulation of beta1-receptor mRNA and protein, upregulation of atrial natriuretic peptide mRNA expression, and increased myocyte diameter indicated similar degrees of failure and hypertrophy in the IDC and PPH phenotypes. The only molecular phenotypic difference between PPH and IDC RVs was upregulation of beta2-receptor gene expression in PPH but not IDC. The major new findings were that (a) both nonfailing intact and explanted human ventricular myocardium expressed substantial amounts of alpha-myosin heavy chain mRNA (alpha-MHC, 23-34% of total), and (b) in heart failure alpha-MHC was downregulated (by 67-84%) and beta-MHC gene expression was upregulated. We conclude that at the mRNA level nonfailing human heart expresses substantial alpha-MHC. In myocardial failure this alteration in gene expression of MHC isoforms, if translated into protein expression, would decrease myosin ATPase enzyme velocity and slow speed of contraction. PMID- 9410911 TI - Selective inhibition of syncytium-inducing and nonsyncytium-inducing HIV-1 variants in individuals receiving didanosine or zidovudine, respectively. AB - By studying changes in the clonal composition of HIV-1 populations during the first weeks of zidovudine (ZDV) treatment before the development of ZDV resistance-conferring mutations, we demonstrated previously a selective inhibition of nonsyncytium-inducing (NSI) HIV-1, even when present as coexisting population in individuals also harboring syncytium-inducing (SI) HIV-1. In this study, we observed the opposite in individuals receiving didanosine (ddI) treatment. In these individuals (n = 7) a median -0.98 log change (range -1.55 0.08) in infectious cellular SI load was observed, whereas the coexisting NSI load was only minimally affected (median -0.15 log, range -1.27-0.50; P = 0.03). The virus phenotype-dependent treatment responses were independent of the clonal composition of HIV-1 populations at baseline. Individuals treated with a combination of ZDV and ddI revealed an equal decline of both NSI and SI infectious cellular load (n = 4; NSI: median -1.55 log, range -2.19 to -1.45; SI: median -1.47 log, range -1.81 to -0.86; P = 0.56). To test the hypothesis that the previously reported optimal activation of ZDV and ddI in activated and resting T cells, respectively, in combination with the differential T cell tropism of NSI and SI HIV-1 is the basis for the observed virus phenotype specific efficacy of nucleoside analogs, we studied the effect of treatment with a protease inhibitor that does not require intracellular activation. Individuals receiving ritonavir (n = 4) indeed showed equal declines in NSI and SI infectious cellular load (NSI: median -2.37 log, range -2.59 to -2.16; SI: median -2.82 log, range -3.14 to -2.50; P = 0.25). Our data suggest HIV-1 phenotype as an additional parameter in the design of optimal treatment regimens. PMID- 9410912 TI - Binding of beta-amyloid to the p75 neurotrophin receptor induces apoptosis. A possible mechanism for Alzheimer's disease. AB - Alzheimer's disease is a neurodegenerative disorder characterized by the extracellular deposition in the brain of aggregated beta-amyloid peptide, presumed to play a pathogenic role, and by preferential loss of neurons that express the 75-kD neurotrophin receptor (p75NTR). Using rat cortical neurons and NIH-3T3 cell line engineered to stably express p75NTR, we find that the beta amyloid peptide specifically binds the p75NTR. Furthermore, 3T3 cells expressing p75NTR, but not wild-type control cells lacking the receptor, undergo apoptosis in the presence of aggregated beta-amyloid. Normal neural crest-derived melanocytes that express physiologic levels of p75NTR undergo apoptosis in the presence of aggregated beta-amyloid, but not in the presence of control peptide synthesized in reverse. These data imply that neuronal death in Alzheimer's disease is mediated, at least in part, by the interaction of beta-amyloid with p75NTR, and suggest new targets for therapeutic intervention. PMID- 9410913 TI - Germ cell apoptosis after treatment of cryptorchidism with human chorionic gonadotropin is associated with impaired reproductive function in the adult. AB - Cryptorchidism results in impaired fertility. Reduced numbers of testicular germ cells can be shown histologically during the first years of life. The process causing germ cell loss in cryptorchid prepubertal boys is unknown, but it could be the result of a form of programmed cell death known as apoptosis. 25 adult men with a history of surgically treated cryptorchidism were studied, 15 of whom had received an unsuccessful human chorionic gonadotropin (hCG) therapy before orchidopexy. Apoptotic DNA fragmentation was assayed in testis biopsies taken during orchidopexy by end-labeling, both in extracted DNA and histochemically in situ. Only a few scattered apoptotic spermatogonias were seen by end-labeling of biopsies from patients not treated with hCG, whereas more extensive labeling of spermatogonia was seen after hCG treatment. As estimated by gel electrophoresis, the amount of low molecular weight DNA was 4.3-fold higher in the hCG-treated group when compared with the level in scrotal testis of non-hCG-treated patients (P < 0.001). About 20 yr after the biopsy, the low molecular weight DNA fragmentation correlated negatively with the testis volume (r = -0.84; P < 0.001) and positively with serum FSH levels (r = 0.73; P < 0.001). Findings in the semen analysis were similar between the groups. Apoptotic loss of spermatogonia after hCG treatment of cryptorchidism warrants reevaluation of the safety of this treatment. PMID- 9410914 TI - Hypoxia inhibits gene expression of voltage-gated K+ channel alpha subunits in pulmonary artery smooth muscle cells. AB - Activity of voltage-gated K+ channels (KV) in pulmonary arterial smooth muscle cells (PASMC) is pivotal in controlling membrane potential, cytoplasmic free Ca2+ concentration ([Ca2+]cyt, and pulmonary vasomotor tone. Acute hypoxia selectively inhibits KV channels, depolarizes PASMC, raises [Ca2+]cyt, and causes pulmonary vasoconstriction and vascular remodeling. Prolonged hypoxia (24-60 h) decreased significantly the mRNA levels of KV channel alpha subunits, KV1.2 and KV1.5. Consistently, the protein levels of KV1.2 and KV1.5 were also decreased significantly by hypoxia (48-72 h). Nevertheless, hypoxia affected negligibly the mRNA levels of KV channel beta subunits (KVbeta1, KVbeta2, and KVbeta3). The native K+ channels are composed of pore-forming alpha and auxiliary beta subunits. Assembly of KV beta subunits with alpha subunits confers rapid inactivation on the slowly or non-inactivating delayed rectifier KV channels. KV beta subunits also function as an open-channel blocker of KV channels. Thus, the diminished transcription and expression of KV alpha subunits may reduce the number of KV channels and decrease KC currents. Unchanged transcription of KV beta subunits may increase the fraction of the KV channel alpha subunits that are associated with beta subunits and further reduce the total KV currents. These data demonstrate a novel mechanism by which chronic hypoxia may cause pulmonary vasoconstriction and hypertension. PMID- 9410915 TI - Evidence for decreased splanchnic glucose uptake after oral glucose administration in non-insulin-dependent diabetes mellitus. AB - The role of splanchnic glucose uptake (SGU) after oral glucose administration as a potential factor contributing to postprandial hyperglycemia in non-insulin dependent diabetes mellitus (NIDDM) has not been established conclusively. Therefore, we investigated SGU in six patients with NIDDM and six weight-matched control subjects by means of the hepatic vein catheterization (HVC) technique. In a second part, we examined the applicability of the recently developed OG-CLAMP technique in NIDDM by comparing SGU and first-pass SGU during HVC with SGU during the OG-CLAMP experiment. The OG-CLAMP method combines a euglycemic, hyperinsulinemic clamp and an oral glucose tolerance test (75 g) during steady state glucose infusion (GINF). During HVC, SGU equals the splanchnic fractional extraction times the total (oral and arterial) glucose load presented to the liver. For OG-CLAMP, SGU was calculated as first-pass SGU by subtracting the integrated decrease in GINF over 180 min from 75 g. Cumulative splanchnic glucose output after oral glucose correlated significantly between both methods and was increased significantly in NIDDM patients (73.1+/-5.1 g for HVC, 76.5+/-5.5 for OG-CLAMP) compared with nondiabetic patients (46.7+/-4.4 g for HVC, 57.5+/-1.9 for OG-CLAMP). Thus, in NIDDM patients, SGU (7.4+/-2.1 vs. 37.8+/-5.9% in nondiabetic patients, P < 0.001) and first-pass SGU (4.7+/-1.7 vs. 26.5+/-5.1% in nondiabetic patients, P < 0.01) were decreased significantly during HVC, as was SGU during OG-CLAMP (3.9+/-1.7 vs. 23.4+/-2.5% in nondiabetic patients, P < 0.0001). SGU measured during OG-CLAMP correlated significantly with SGU (r = 0.87, P < 0.05 for NIDDM patients; r = 0.94, P < 0.01 for nondiabetic patients) and first-pass SGU (r = 0.87, P < 0.05 for NIDDM patients; r = 0.84, P < 0.05 for nondiabetic patients) during HVC. In conclusion, (a) SGU after oral glucose administration is decreased in NIDDM as measured by both methods, and (b) SGU during the OG-CLAMP is well-correlated to SGU and first-pass SGU during HVC in NIDDM. The decrease in SGU in NIDDM might contribute to postprandial hyperglycemia in diabetic subjects. PMID- 9410916 TI - Myosin heavy chain gene expression in human heart failure. AB - Two isoforms of myosin heavy chain (MyHC), alpha and beta, exist in the mammalian ventricular myocardium, and their relative expression is correlated with the contractile velocity of cardiac muscle. Several pathologic stimuli can cause a shift in the MyHC composition of the rodent ventricle from alpha- to beta-MyHC. Given the potential physiological consequences of cardiac MyHC isoform shifts, we determined MyHC gene expression in human heart failure where cardiac contractility is impaired significantly. In this study, we quantitated the relative amounts of alpha- and beta-MyHC mRNA in the left ventricular free walls (LVs) of 14 heart donor candidates with no history of cardiovascular disease or structural cardiovascular abnormalities. This group consisted of seven patients with nonfailing (NF) hearts and seven patients with hearts that exhibited donor heart dysfunction (DHD). These were compared with 19 patients undergoing cardiac transplantation for chronic end-stage heart failure (F). The relative amounts of alpha-MyHC mRNA to total (i.e., alpha + beta) MyHC mRNA in the NF- and DHD-LVs were surprisingly high compared with previous reports (33.3+/-18.9 and 35.4+/ 16.5%, respectively), and were significantly higher than those in the F-LVs, regardless of the cause of heart failure (2.2+/-3.5%, P < 0.0001). There was no significant difference in the ratios in NF- and DHD-LVs. Our results demonstrate that a considerable amount of alpha-MyHC mRNA is expressed in the normal heart, and is decreased significantly in chronic end-stage heart failure. If protein and enzymatic activity correlate with mRNA expression, this molecular alteration may be sufficient to explain systolic dysfunction in F-LVs, and therapeutics oriented towards increasing alpha-MyHC gene expression may be feasible. PMID- 9410917 TI - DNA immunization of neonates induces immunity despite the presence of maternal antibody. AB - Neonatal animals were not considered as suitable vaccine recipients either because of immune immaturity or because passively delivered antibody interferes with immune induction. In this report, we evaluated the response of neonatal mice to immunization with naked DNA encoding a herpes simplex virus (HSV) protein, and determined if maternally derived HSV antibody interfered with immunogenicity. Our results show that neonatal mice develop effective humoral and T cell responses after immunization with either DNA or inactivated vaccines. The nature of the responses to HSV immunization, however, was more Th2-like in neonates than in adults. Whereas neonatal mice from HSV-naive mothers responded well to both DNA and inactivated vaccines, only DNA immunization induced effective immunity in neonates born to immune mothers. Our results indicate that DNA vaccines might provide a useful means of immunizing young animals that still possess high levels of potentially interfering maternal antibody. PMID- 9410918 TI - Immunological significance of cytotoxic T lymphocyte epitope variants in patients chronically infected by the hepatitis C virus. AB - This study was performed to test the hypothesis that cytotoxic T lymphocyte (CTL) selection of hepatitis C virus (HCV) escape variants plays a role in HCV persistence. The peripheral blood CTL responsiveness of patients with well established chronic hepatitis C to a panel of 10 prototype HCV peptides (genotype 1a) was compared with the corresponding sequences encoded by the infecting viruses in each patient. Variant viral peptide sequences were threefold more frequent in the presence of a CTL response than in its absence, and CTL responses were detected nearly twice as often in association with variant rather than with prototype viral peptide sequences. Furthermore, over half of the patients were infected with potential CTL escape variants that contained nonimmunogenic and noncross-reactive variant peptides many of which displayed reduced HLA-binding affinity. Surprisingly, follow up analysis over a period of up to 46 mo revealed that, in contrast to the relatively high frequency of escape variants initially observed, the subsequent emergence rate of CTL escape variants was very low. Interestingly, the one escape variant that was detected proved to be a CTL antagonist. Collectively, these observations suggest that CTL selection of epitope variants may have occurred in these patients before their entrance into the study and that it may have played a role in HCV persistence. The low apparent rate of ongoing CTL selection in chronically infected patients, however, suggests that if CTL escape occurs during HCV infection it is probably an early event. PMID- 9410919 TI - Somatostatin receptor (SSTR) subtype-selective analogues differentially suppress in vitro growth hormone and prolactin in human pituitary adenomas. Novel potential therapy for functional pituitary tumors. AB - Previously, we have shown somatostatin receptor (SSTR) subtype-specific regulation of growth hormone (GH), thyroid-stimulating hormone, and prolactin (PRL) secretion in human fetal pituitary cultures, where GH and thyroid stimulating hormone are mediated by both SSTR2 and SSTR5, whereas SSTR2 preferentially mediates PRL secretion. We now tested SSTR subtype-selective analogues in primary human GH- and PRL-secreting pituitary adenoma cultures. Analogue affinities determined by membrane radioligand binding in cells stably expressing human SSTR forms were either SSTR2 or SSTR5-selective. Analogues preferential either for SSTR2, including octreotide, lanreotide, and novel compounds with improved affinity for SSTR2, or new SSTR5-selective compounds suppressed GH in tumor cell cultures (up to 44% of control; P < 0.0005). However, novel analogues from both groups were 30-40% more potent than octreotide and lanreotide in suppressing GH (P < 0.05). Heterologous analogue combinations containing both SSTR2- and SSTR5-selective compounds were more potent in decreasing GH than analogues used alone (P < 0.05), or than combinations of compounds specific for the same receptor subtype (P < 0.005). In contrast, SSTR2 selective analogues did not suppress PRL release from six cultured prolactinomas studied. However, new SSTR5-selective analogues suppressed in vitro PRL secretion (30-40%; P < 0.05) in four of six prolactinomas. These results suggest that both SSTR2 and SSTR5 are involved in GH regulation in somatotroph adenoma cells, whereas SSTR5 exclusively regulates PRL secretion from prolactinoma cells. Thus, somatostatin analogues with improved selective binding affinity for these receptor subtypes may be effective in the treatment of either GH- or PRL secreting adenomas. PMID- 9410921 TI - Self-assessment examination of the American Academy of Dermatology. Necrolytic migratory erythema, erythema gyratum repens and calciphylaxis. PMID- 9410922 TI - Energy and nutrient inadequacies in the diets of low-income women who breast feed. AB - OBJECTIVE: To assess the energy and nutrient intakes of women who are breast feeding in relation to the Recommended Dietary Allowances (RDAs) for energy and nutrients during lactation. DESIGN: Survey using a interviewer-administered questionnaire and a 24 dietary recall. SUBJECTS: The subjects were 183 women breast-feeding at 3 months postpartum. All were living in low-income communities in Ontario, Canada, that were participating in the longitudinal prevention initiative, Better Beginnings, Better Futures. STATISTICAL ANALYSES: Mann-Whitney U test to compare energy and nutrient intakes of women with incomes above and below the Statistics Canada poverty line. RESULTS: Median intake of energy (2,148 kcal) was below the RDA of 2,700 kcal. Compared with the RDAs, intakes of calcium (928 vs 1,200 mg), folate (222 vs 280 microns), iron (13 vs 15 mg), thiamin (1.4 vs 1.6 mg), vitamin A (846 vs 1,300 retinol equivalents), and zinc (10 vs 19 mg) were below recommended values for women who are lactating. Although household income for 70% of the women was below the poverty line, differences in energy and nutrient intakes according to income group were not statistically significant. APPLICATIONS: Lactating women are at high risk of energy and nutrient inadequacies. Programs to increase breast-feeding rates, particularly among low income communities, must include strategies to ensure adequate diets during lactation. PMID- 9410923 TI - Audrey C. Wright, MS, RD, receives 1997 Copher Memorial Award. PMID- 9410924 TI - [Real impact of prematurity]. PMID- 9410920 TI - Functional CD40 ligand is expressed by T cells in rheumatoid arthritis. AB - CD40 ligand (CD40-L), a member of the tumor necrosis family of transmembrane glycoproteins, is rapidly and transiently expressed on the surface of recently activated CD4+ T cells. Interactions between CD40-L and CD40 induce B cell immunoglobulin production as well as monocyte activation and dendritic cell differentiation. Since these features characterize rheumatoid arthritis (RA), the expression and function of CD40-L in RA was examined. Freshly isolated RA peripheral blood (PB) and synovial fluid (SF) T cells expressed CD40-L mRNA as well as low level cell surface CD40-L. An additional subset of CD4+ RA SF T cells upregulated cell surface CD40-L expression within 15 min of in vitro activation even in the presence of cycloheximide, but soluble CD40-L was not found in SF. CD40-L expressed by RA T cells was functional, since RA PB and SF T cells but not normal PB T cells stimulated CD40-L-dependent B cell immunoglobulin production and dendritic cell IL-12 expression in the absence of prolonged in vitro T cell activation. In view of the diverse proinflammatory effects of CD40-L, this molecule is likely to play a central role in the perpetuation of rheumatoid synovitis. Of importance, blockade of CD40-L may prove highly effective as a disease modifying therapy for RA. PMID- 9410925 TI - [Premature labor: treatment objectives]. PMID- 9410926 TI - [Cervical ripening]. PMID- 9410927 TI - [Hemorrhage during delivery: management in France and value of prostaglandins]. PMID- 9410928 TI - [Postpartum hemorrhage: perspectives in the United States]. PMID- 9410929 TI - [Classification of the risks of premature labor. Implications for treatment]. PMID- 9410930 TI - A review of peer review. PMID- 9410931 TI - [Collagen diseases: new diagnostic methods and progress in treatment]. PMID- 9410932 TI - [Progress in the concept and diagnosis of collagen diseases. 1. Transition and changes]. PMID- 9410933 TI - [Progress in the concept and diagnosis of collagen diseases. 2. New concept of immunologic anomalies]. PMID- 9410934 TI - [Progress in the concept and diagnosis of collagen diseases. 3. New tests and key points in their application]. PMID- 9410935 TI - [Visceral lesions and progress in the diagnosis of collagen diseases. 1) Kidney dysfunctions]. PMID- 9410936 TI - [Organ lesions and progress in diagnosis of collagen diseases. 2) Pulmonary lesions]. PMID- 9410937 TI - [Organ lesions and progress in diagnosis of collagen diseases. 3) Lesions of digestive organs]. PMID- 9410938 TI - [Organ lesions and progress in diagnosis of collagen diseases. 4) Lesions of the nervous system]. PMID- 9410939 TI - [Organ lesions and progress in diagnosis of collagen diseases. (5) Cardiovascular lesions]. PMID- 9410940 TI - [Concept and management of specific physiopathology of collagen diseases. 1) Approach to healthy individuals with positive anti-nuclear antibody reactions]. PMID- 9410941 TI - [Concept of specific physiopathology and management of collagen diseases. 2) Approach to anti-phospholipid antibody syndrome]. PMID- 9410942 TI - [Concept of specific physiopathology and management of collagen diseases. 3) Approach to SLE in pregnancy]. PMID- 9410943 TI - [Current treatment of collagen diseases. 1. Glucocorticoids: key points in their application]. PMID- 9410944 TI - [Current treatment of collagen diseases. 2. Non-steroidal anti-inflammatory agents: key points in their application]. PMID- 9410945 TI - [Current treatment of collagen diseases. 3. Antirheumatic agents: key points in their application]. PMID- 9410946 TI - [Current treatment of collagen diseases. 4. Immunosuppressive agents: key points in their application]. PMID- 9410947 TI - [Current treatment of collagen diseases. 5. Self care and QOL]. PMID- 9410948 TI - [Collagen diseases : new diagnostic methods and progress in treatment. Discussion]. PMID- 9410949 TI - [Case of chronic pancreatitis discovered at the development of malabsorption syndrome]. PMID- 9410950 TI - [Autopsy case of infection-associated hemophagocytic syndrome caused by E. coli induced acute pyelonephritis]. PMID- 9410951 TI - [Autopsy case of primary amyloidosis developing refractory ascites and undergoing rapid deterioration]. PMID- 9410952 TI - [Case of low-grade fibromyxoid sarcoma]. PMID- 9410953 TI - [Deviation in CD28 expression by CD8-positive T cells noted in virus infections]. PMID- 9410954 TI - [Progress in molecular biological study of constitutional jaundice]. PMID- 9410955 TI - [Myofibroblastosis--importance of transformation of renal cells into myofibroblasts during the renal fibrosis process]. PMID- 9410956 TI - [Human, nature and medicine--future internal medicine]. PMID- 9410957 TI - [External factor and the nervous system]. PMID- 9410958 TI - [Platelet dysfunction--from bedside clinical study to molecular study]. PMID- 9410959 TI - [Physiopathology and therapy of hypertension]. PMID- 9410960 TI - [Liver diseases and nutritional therapy]. PMID- 9410961 TI - [Ideal sugar level control and diabetic angiopathies]. PMID- 9410962 TI - [Apoptosis and diseases]. PMID- 9410963 TI - [Malignant tumor and apoptosis]. PMID- 9410964 TI - [Autoimmunity and apoptosis]. PMID- 9410965 TI - [Specific suppression of arthritis rheumatoid by Fas/Fas ligand]. PMID- 9410966 TI - [Hematological disorders and apoptosis]. PMID- 9410967 TI - [Liver diseases and apoptosis]. PMID- 9410968 TI - [Nervous system diseases and apoptosis]. PMID- 9410969 TI - [Endocrine diseases and apoptosis]. PMID- 9410970 TI - [Thrombolytic therapy of symptomatic / asymptomatic cerebral infarction]. PMID- 9410971 TI - [Thrombolytic therapy of ischemic heart disease]. PMID- 9410972 TI - [Thrombolytic therapy of ischemic heart disease]. PMID- 9410973 TI - [Thrombolytic therapy of multiple thromboembolism-- atrial fibrillation]. PMID- 9410974 TI - [Thrombolytic therapy of vasculitis and thrombotic microangiopathy--with special reference to the kidney]. PMID- 9410975 TI - [Nephrotic syndrome and the thrombolytic therapy]. PMID- 9410976 TI - [Disorders of body water regulation and the therapy--water metabolism disorder and vasopressin]. PMID- 9410977 TI - [Disorders of body water regulation and the therapy--aquaporin and water metabolism disorders]. PMID- 9410979 TI - [Disorders of body water regulation and the therapy using diuretics]. PMID- 9410978 TI - [Disorders of body water regulation and the therapy--vasopressin antagonists]. PMID- 9410980 TI - [Disorders of body water regulation and the therapy--acid-base equilibrium in the kidney and disorders]. PMID- 9410981 TI - [Disorders of body water regulation and the therapy--role of atrial natriuretic peptide in homeostasis of body water in heart failure]. PMID- 9410982 TI - [Ideal of multicenter clinical trials--the scientific and ethical aspects - experience in epidemiological study of Hisayama town in Japan]. PMID- 9410984 TI - [Ideal of multicenter clinical trials--the scientific and ethical aspects- clinical trials on nervous system disease]. PMID- 9410983 TI - [Ideal of multicenter clinical trials--the scientific and ethical aspects- clinical trials on cardiovascular disease]. PMID- 9410985 TI - [Ideal of multicenter clinical trials--the scientific and ethical aspects- clinical trials on cancer]. PMID- 9410986 TI - [Ideal of multicenter clinical trials--the scientific and ethical aspects--point of view from the Department of Health and Welfare]. PMID- 9410987 TI - [ANCA related nephritis]. PMID- 9410988 TI - [Diagnostic imaging of dementia]. PMID- 9410989 TI - [Autologous and homologous peripheral blood stem cell transplantation]. PMID- 9410991 TI - [Home oxygen inhalation therapy--management of respiration during sleep]. PMID- 9410990 TI - [Clinical study on eating behavior disorders]. PMID- 9410992 TI - [Management of diabetic nephropathy]. PMID- 9410993 TI - [Progress on coronary artery intervention therapy]. PMID- 9410994 TI - [Etiology and therapy of thyroid function disorders]. PMID- 9410995 TI - [Progress on therapy and etiological study of bronchial asthma]. PMID- 9410997 TI - [Physiopathology and therapy of G-type hepatitis]. PMID- 9410996 TI - [Physiopathology and molecular mechanism of therapy related leukemia]. PMID- 9410998 TI - [Drug therapy of congestive heart failure]. PMID- 9410999 TI - [New strategy for immunity regulation]. PMID- 9411000 TI - [Diabetes mellitus and the gene]. PMID- 9411001 TI - [Clinical significance of adrenomedullin]. PMID- 9411002 TI - [Respiratory disorders and nitric oxide]. PMID- 9411003 TI - [Asymptomatic cerebral infarction]. PMID- 9411004 TI - [Recent trends on study of AIDS]. PMID- 9411005 TI - Basal cell carcinoma of the external auditory canal and Gorlin-Goltz syndrome: a case report. AB - A case of nevoid basal cell carcinoma syndrome (Gorlin-Goltz syndrome) with basal cell carcinoma of the external auditory canal is reported. Thus is only the second such case. PMID- 9411006 TI - [Calcium antagonists in ischemic heart disease]. AB - Recently, there has been some controversy concerning calcium antagonists, suggesting the need for further debate on this heterogeneous class of drugs. Three chemical families, dihydropyridines (DHP), phenylalkylamines (verapamil) and benzothiazepines (diltiazem) bind to the type L receptors of the calcium channels with different binding, modulation and tissue selectivity characteristics. DHP are selective for type L receptors and block the extracellular portion of the channel leading to vigorous vasodilatation and little or no cardiodepressive effect. Diltiazem and verapamil also interfere with type T channel receptors. These drugs have a cardiodepressive and a bradycardia effect. Verapamil blocks the intracellular portion of the calcium channel at the site where part of the catecholamine effect occurs, leading to less reflex sympathetic activation than with other calcium antagonists (namely DHP). Deleterious sympathetic stimulation is proportional to the intensity and degree of rapid onset of arterial vasodilatation and is attenuated with slow-release formulations. Calcium antagonists in general have an anti-angina effect but high dose short-acting DHP can cause excessive vasodilatator leading to subsequent ischemia. In chronic stable angina, slow-release verapamil has been shown to have a preventive clinical effect comparable to that of beta blockers. Slow-release nifedipine is effective and safe but must be associated with betablockers. In unstable angina, only those calcium antagonists with a bradycardia effect appear to have an effect similar to beta blockers. Beta blockers are nevertheless to be preferred in these patients (excepting Prinzmetal angina) until results of convincing clinical studies are available. After the initial phase of myocardial infarction, again only calcium antagonists with a bradycardia effect have been shown to have a beneficial effect, in patients without cardiac failure: diltiazem in infarction without Q-wave and verapamil in all infarctions, in case of residual ischemia to reduce the risk of recurrence. PMID- 9411007 TI - [Malingering in vascular disease]. AB - Pathomimicry occurs in all fields of medicine. Although difficult to recognize, all physicians should be aware of the underlying mechanisms in order to avoid excessive ordering of complementary examinations and therapeutic propositions which may be dangerous. Pathomimesis is to be distinguished from Munchhausen's syndrome which involves simulation of severe disease and extravagant lies with false history reporting leading to successive hospitalizations in different hospitals. Pathomimesis is also distinguished by the goal of the simulation which is to obtain a precise material benefit. In vascular pathology, pathomimesis can take on several aspects:hemorrhagic syndrome by self-prescribed anticoagulants, self-induced limb edema (tourniquet), or self-inflicted skin wounds. Diagnosis is suggested by the absence of a cause, identification of the stricture groove in case of edema, imprivement with occlusive dressings for skin ulcers and by the general presentation. Pathomimesis is usually encountered in young intelligent women with some medical knowledge. This behavior has a psychopathological significance, the provoked symptoms demonstrating difficult emotional events in the past. The patient attempts to overcome an earlier tragic situation. Pathomimicry is thus expressed during acute episodes of fear and/or anxiety. For the practitioner, it is important to avoid accusing the patient or attempting to get the patient to avow as there is an important risk of exaggerated or self destructive response. The patient should be led to realize that the physician knows what is happening. This unstated interchange allows the patient to establish a confident relationship with the physician, a relationship which should lead to an accepted psychotherapy. PMID- 9411008 TI - [Venous thrombosis of the legs and cancer. Evaluation of risk factors of venous thrombosis in the medical environment]. AB - In medical patients, risk factors of leg venous thrombosis are not well evaluated. Cancer is considered as an important one. The aim of this study was to evaluate the role of intrinsic thrombotic risk (tumor hypercoagulable state) and external thrombotic risk (associate factors). We have made a prospective analysis of thrombotic venous risk factors in two medical populations with leg venous thrombosis: patients with cancer and patients without cancer. Risk of thrombosis depends on the thrombogenic importance of the risk factor and its chronicity or not. We assessed cancer and thromboembolic disease at the time of diagnosis and during a median follow up of 125.2 days. We included 31 consecutive cases of cancer (21 men, 10 women, mean age 63.8 years), and 50 consecutive cases of non cancer patients (32 men, 18 women, mean age 65.5 years), these two populations were not different. The classic risk factors of venous thrombosis were not frequent in cancer patient. Analysis of thrombotic risk showed that 61% of cancer patient group had venous thrombosis without classic thrombotic risk, as compared to 32% in non cancer patient group, showing the direct role of cancer in thrombosis (p < 0.01). The cancer was often aggressive and metastatic adenocarcinoma of various origins. The effect of chemotherapy is not clear, only hormonotherapy seemed to be responsible in two cases. Cancer hypercoagulability, defined by clinical characteristics, is a real risk factor of venous thrombosis but of low frequency. Indeed, the incidence of venous thrombosis in oncologic unit is rare (0.4%). Finally, thromboembolic disease in cancer patients is not different than in no cancer patients, Trousseau's syndrome is unfrequent. Prognosis is poor (40% death with 44.5 days of median survival), and antithrombotic therapy complications are frequent (bleeding 16%, oral anticoagulants resistance 20%). PMID- 9411009 TI - [Effect of smoking on blood rheology]. AB - The aim of this study was to compare the effects of cigarette smoking on biologic and rheologic tests, chiefly on the red blood cells (RBC) in measuring the deformability by the Cell Transit Analyser (CTA) and their aggregation by using an ultrasonic interferometry method based on A-mode echography allowed for the measurement of the accumulation rate of particles in a solid plate which is related to their sedimentation rate (Echo-Cell). Nine male smoker subjects with a high nicotine addiction measured by Fagerstrom questionnaire (> 8) and level of carbon monoxide (CM) in the breathed out air (> 20 ppm), have been compared with ten healthy no-smoker volunteers (CM < 3 ppm). One smoker has been eliminated of statistic evaluations because his glucose level showed a diabetes (10.5 mmol/l). A nailfold capillaroscopy performed in all subjects has eliminated the patterns of latent vasculitis or scleroderma. RBC and platelets counts, hemoglobin, ionogram, gamma GT, ASAT, ALAT, uric acid, total cholesterol and glucose levels were not significantly different between the two groups. On the other hand, in the smoker group, white blood cells count, serum triglycerides and especially fibrinogen values were higher than in the non-smoker's group. RBC sedimentation rate was normal in the two groups but was higher in smoker's group too. Without consumption of alcohol, the mean RBC volume was more important in smokers (91.9 +/- 1.2 versus 87.5 +/- 0.4, p = 0.003). Rheologic tests were more pathologic in smokers. The transit time or RBC by CTA was longer than in control group (1.6 ms +/- 0.02 versus 1.2 +/- 0.05, p = 0.0003). Echo-Cell technic showed a number and size of RBC aggregates more important with a rate of speed of accumulation higher than in the control group. These results demonstrated the toxic effects of smoking alone on blood toward a propensity for thrombotic status. PMID- 9411010 TI - [Effect of meteorological variations on the emergence of deep venous thrombosis of the leg]. AB - INTRODUCTION: Recent articles have established a significant relationship between metereology variables and the development of vascular disease. We performed a retrospective study to determine relationships between the development of deep vein thrombosis in the lower limb and certain meteorology variables. MATERIALS AND METHODS: We identified 345 cases of phlebitis in 1995. We studied the distribution of the number of venous thrombosis per day, per month and per season. We compared certain meteorological data (atmospheric pressure, temperature, mean hygrometery) for days with and days without venous thrombosis and the atmospheric variations during the 48 hours prior to venous thrombosis. RESULTS: There was a significant relationship (p < 0.004) between the mean number of cases of phlebitis recorded per day and season with winter predominating. On days when phlebitis occurred, the atmospheric pressure was significantly lower (p < 0.05). The number of thrombotic events was significantly different on days when the variation was greater than 10 hectopascals than on days when the variation was less than 10 hectopascals (p < 0.05). CONCLUSIONS: In our study, deep vein thrombosis of the lower limb was significantly associated with certain meteorology variables. Prospective multicentric studies are needed to confirm these relationships. PMID- 9411011 TI - [Medical recommendations and references about hypolipidemic drugs. ANDEM (National Agency for the Development of Medical Evaluation]. PMID- 9411012 TI - [Pulmonary embolism and unusual deep venous thrombosis. Report of two cases]. AB - Contrast venography is the gold standard for the diagnosis of deep vein thrombosis in the lower limb extremities, but it fails to visualize deep veins like deep femoral vein and internal iliac vein. The internal iliac can be examined with duplex scanning if the technique and the examination conditions are correct. As reported in these two cases, thrombosis of these deep veins may lead to pulmonary embolism. The first case is a young female with venous thromboembolic disease in whom internal iliac vein thrombosis was documented only at the second examination. In the second case, deep femoral vein thrombosis appeared early in a comatose young male. This thrombosis may be classified as proximal muscular vein thrombosis. These two cases emphasize the importance of a duplex scanning examination performed with rigorous technique, whose the main limitation being examination conditions. PMID- 9411013 TI - [Non-invasive management of a serious acute pulmonary embolism]. AB - Prompt diagnosis of a large pulmonary embolus is essential in order to initiate appropriate treatment early. We report a case of a large pulmonary embolus in which management was aided solely by noninvasive investigations. Transthoracic echocardiogram showed elevated right heart pressures which together with the patient symptoms suggested a major pulmonary embolus. Spiral computed tomography of the chest confirmed the diagnosis. The source of the embolus was shown by echodoppler. This case illustrates that a diagnosis of a major pulmonary embolus can be made using noninvasive techniques. Pulmonary angiography should be reserved for those rare cases in which diagnostic uncertainty remains rather than being used as a routine examination prior to consideration of therapeutic decision. PMID- 9411014 TI - [Post-infectious systemic vasculitis]. PMID- 9411015 TI - 24th International Congress on Electrocardiology and 38th International Symposium on Vectorcardiography. Abstracts. PMID- 9411016 TI - Oligonucleotide adjuvants for T helper 1 (Th1)-specific vaccination. PMID- 9411017 TI - Same-different texture discrimination in pigeons: testing competing models of discrimination and stimulus integration. AB - The choice behavior of 6 pigeons performing a mulltidimensional same-different texture discrimination was examined. On each trial, they had to choose among 2 choice hoppers depending on whether a color, shape, or redundant (color and shape) target signal was present or not in a textured stimulus. Receiver operating characteristic (ROC) curves were produced by variations in the priori signal presentation probabilities across conditions. Quantitative analyses of these ROC curves were used to evaluate different competing theories of discrimination (signal detection vs. high-threshold-default response models) and information integration (independent observations, additive integration, unidimensional models). The results suggested the structure of the pigeons' choice behavior in this same-different discrimination was best described by an unequal variance signal detection model involving a unidimensional evidence variable (e.g., degree of difference). PMID- 9411018 TI - Pigeon same-different concept learning with multiple stimulus classes. AB - Two experiments examined the acquisition and transfer of a complex same-different discrimination by pigeons. With the use of a 2-alternative choice task, 5 pigeons were reinforced for discriminating odd-item Different displays in which a contrasting target ws present, from Same displays, in which all elements were identical. Four different types of same-different displays were concurrently tested. The display types differed in their configuration (texture vs. visual search organization), the nature of their elements (small and large colored shapes; pictures of birds, flowers, fish, and humans), and the processing demands required by their global-local element arrangement. Despite these differences, the pigeons learned to discriminate all 4 display types at the same rate and showed positive discrimination transfer to novel examples of each type, suggesting that a single generalized rule was used to discriminate all display types. These results provide some of the strongest evidence yet that pigeons, like many primates, can learn an abstract, visually mediated same-different concept. PMID- 9411019 TI - Picture fragment completion: priming in the pigeon. AB - It has been suggested that the system behind implicit memory in humans is evolutionarily old and that animals should readily show priming. In Experiment 1, a picture fragment completion test was used to test priming in pigeons. After pecking a warning stimulus, pigeons were shown 2 partially obscured pictures from different categories and were always reinforced for choosing a picture from one of the categories. On control trials, the warning stimulus was a picture of some object (not from the S+ or S- category), on study trials the warning stimulus was a picture to be categorized on the next trial, and on test trials the warning stimulus was a randomly chosen picture and the S+ picture was the warning stimulus seen on the previous trial. Categorization was better on study and test trials than on control trials. Experiment 2 ruled out the possibility that the priming effect was caused by the pigeons' responding to familiarity by using warning stimuli from both S+ and S- categories. Experiment 3 investigated the time course of the priming effect. PMID- 9411020 TI - Spatial sampling of motion: seeing an object moving behind a picket fence. AB - Spatially sampled motion (H.P. Snippe & J.J. Koenderink, 1994) leads to time lagged correlations of luminance change at discrete spatial positions. Observers matched the perceived width of a bar whose motion path was sampled spatially to the width of a static bar; the width of the moving object was not directly observable. Observers did reasonably well on this task when the stimulus onset asynchrony (SOA) between adjacent samples was approximately 90 ms, but performance broke down completely when the SOA was doubled. Performance improved considerably as more samples became available, provided that these samples all fell along the same smooth motion path and were seen by the same eye. This spatiotemporal information in spatially sampled motion can specify the width of a moving object, but it is likely to be useful to observers only if the sampling preserves the impression of motion. PMID- 9411022 TI - Amodal completion of partly occluded surfaces: is there a mosaic stage? AB - Recent investigators have proposed that amodal completion is a sequential process requiring a preliminary mosaic stage. Results of 6 studies of the time course of completion processes show support for this mosaic-first view with pictorial displays, but not with displays involving occlusion specified by binocular parallax or when pictorial displays were observed monocularly. These results still do not rule out the mosaic-first view. A parallel model, however, can account more economically for the available data. PMID- 9411021 TI - The Simon effect occurs relative to the direction of an attention shift. AB - We investigated whether the Simon effect depends on the orienting of attention. In Experiment 1, participants were required to execute left-right discriminative responses to 2 patterns that were presented to the left or right of fixation. The 2 patterns were similar, and the discrimination was difficult. A letter at fixation signaled whether the current trial was a catch trial. The results showed a reversal of the Simon effect. That is, spatially noncorresponding responses were faster than spatially corresponding responses. In Experiment 2, the discrimination of the relevant stimulus attribute was easy. In Experiment 3, the discrimination of the relevant stimulus attribute was difficult, but the stimulus exposure time was long. In either experiment, the regular Simon effect was reinstated. In Experiment 4, the letter that signaled a catch trial appeared to the left or right of the imperative stimulus. The Simon effect occurred relative to the position of the letter. PMID- 9411023 TI - Testing conditions for viewpoint invariance in object recognition. AB - Based on the geon structural description approach, I. Biederman and P.C. Gerhardstein (1993) proposed 3 conditions under which object recognition is predicted to be viewpoint invariant. Two experiments are reported that satisfied all 3 criteria yet revealed performance that was clearly viewpoint dependent. Experiment 1 demonstrated that for both sequential matching and naming tasks, recognition of qualitatively distinct objects became progressively longer and less accurate as the viewpoint difference between study and test viewpoints increased. Experiment 2 demonstrated that for single-part objects, larger effects of viewpoint occurred when there was a change in the visible structure, indicating sensitivity to qualitative features in the image, not geon structural descriptions. These results suggest that the conditions proposed by I. Biederman and P.C. Gerhardstein are not generally applicable, the recognition of qualitatively distinct objects often relies on viewpoint-dependent mechanisms, and the molar features of view-based mechanisms appear to be image features rather than geons. PMID- 9411024 TI - A horse race between independent processes: evidence for a phantom point of no return in preparation of a speeded motor response. AB - Electromyographic (EMG) data show that a speeded elbow extension response can be interrupted at any time after its execution. Submaximal, or partial, EMG data are also observed in some cases, from which 2 alternatives were considered. The partial response might in fact be interrupted early in response production or, alternately, it might arise from stopping processes that incompletely suppress the response production processes prior to their execution. An interrupted response is easily accounted for by a horse race between independent response production and stopping processes, whereas a partial response can only be reconciled if leakage between the two processes is allowed for. If the distinction between an interrupted and a partial response is correct, then the data yielded evidence for a phantom point of no return that locates late in the premotor component of the response and, thus, prior to the onset of EMG activity. PMID- 9411026 TI - Nursing history as a tool for development of a professional identity within nursing students. PMID- 9411025 TI - Clinical and radiologic survivorship of cementless tibial components fixed with finned polyethylene pegs. AB - One hundred twenty patients (22 men, 98 women; 144 knees) with uncemented Freeman Samuelson total knee arthroplasty were followed prospectively. Eighty-one patients had rheumatoid arthritis and 39 patients had osteoarthrosis. The mean follow-up period was 6.8 years. Three different types of tibial components were used: a high-density polyethylene component without stem, a metal-backed tibial component without stem, and a metal-backed tibial component with stem. Progressive varus tilting turned out to be an early sign of failure and occurred in 22% of the tibial components. Revision of the tibial component was done in 17 knees. Survival analysis with revision as endpoint revealed a survival rate of 79% at a follow-up period of 10 years. Cementless fixation of this design using macrointerlocking pegs and no other stabilization resulted in poor fixation and a high revision rate and cannot be recommended. PMID- 9411028 TI - [Imaging diagnosis of parotid gland tumors]. PMID- 9411027 TI - Diet modifies elastase I and II activities and mRNA levels during postnatal development and weaning in piglets. AB - Little information is available on the expression of pancreatic elastase I and II, despite their role in protein milk digestion. We studied the developmental changes and the effects of diet composition on both elastase I and II expression in suckled and weaned piglets. We measured their activities and levels of their corresponding mRNA. Forty-two piglets were assigned to seven groups according to age and diet. Piglets were slaughtered at birth (Group 1), or suckled up to 13 d (Group 2) or 21 d (Group 3), fed a milk substitute from 14 to 21 d (Group 4) or to 56 d (Group 7), suckled up to 21 d and then fed a dry starter up to 56 d (Group 5), or fed a milk substitute from 14 to 21 d and then a dry starter up to 56 d (Group 6). At 21 d pancreatic function was not modified by the source and the form of milk consumed. The specific activity of elastase II was maximum at birth and declined sharply thereafter, whereas that of elastase I markedly increased after weaning. The presence of milk protein in the diet did not prevent the sharp decrease in elastase II activity observed with age. During the 13 d period of suckling sow's milk, the mRNA patterns indicated that the regulation was at the mRNA and post-transcriptional levels, whereas after weaning and depending on the source of dietary protein, it was essentially translational and/or post-translational. Taken together, our results provide evidence of the early expression of elastase I and II genes that could enhance protein digestion. It seems that elastase II might be a predominant pancreatic protease during the milk-feeding period, whereas elastase I might be related to weaning. PMID- 9411029 TI - Consultation section. Refractive surgical problem. PMID- 9411030 TI - Excitatory amino acid receptor antagonists block the cardiovascular and anxiety responses elicited by gamma-aminobutyric acidA receptor blockade in the basolateral amygdala of rats. AB - Blockade of gamma-aminobutyric acid (GABAA) receptors in the anterior basolateral amygdala (BLA) with bicuculline methiodide results in an increase in heart rate, blood pressure and "anxiety" in rats. Glutamate receptors in the BLA are also reported to be involved in eliciting anxiety responses. The purpose of this study was to investigate the interaction between GABAergic inhibition and glutamatergic excitation in the BLA. Male Wistar rts were implanted with femoral arterial catheters and bilateral chronic microinjection cannulae into the BLA. Each animal was injected with either artificial cerebrospinal fluid (100 nl), bicuculline methiodide (10 pmol/100 nl) or bicuculline methiodide + one dose of an antagonist of either the N-methyl-D-aspartate receptor [AP5 (20 and 100 pmol) and dizocilpine (25 and 125 pmol)] or the non-N-methyl-D-aspartate ionotropic receptor [CNQX (10 and 50 pmol) and GYKI 52466 (50 and 250 pmol)]. Increases in heart rate, blood pressure and "anxiety" (as measured in the social interaction test) observed in rats after bicuculline methiodide injections into the BLA were blocked in a dose dependent manner with the concurrent injections of either N methyl-D-aspartate or non-N-methyl-D-aspartate antagonists, suggesting that activation of both subtypes of glutamate ionotropic receptors may be necessary for the responses elicited by GABAA receptor blockade in the basolateral amygdala. PMID- 9411031 TI - Cytomegalovirus retinitis treatment. Abstract and commentary. PMID- 9411032 TI - Immune restoration following treatment. Abstract and Commentary. PMID- 9411033 TI - Quality of care. PMID- 9411034 TI - A JAMA theme issue on quality of care. A new proposal and a call to action. PMID- 9411035 TI - Primer on allergic and immunologic diseases. PMID- 9411036 TI - [Insertion characteristics and complications of a new spinal needle 26-gauge Atraucan]. AB - We evaluated the insertion characteristics and complications of a new spinal needle 26-gauge Atraucan (group A) compared with 27-gauge Whitacre (group W) in 100 patients undergoing orthopedic surgeries of the lower extremities. Spinal anesthesia was performed in the lateral decubitus position and 0.40-0.5% tetracaine 1.6-2.5 ml was injected through the L 3/4 or L 4/5 interspace. The tactile appreciation of dural presentation with the needle (dural click) was higher in group W (89.8%) than in group A (42.6%) which the back flow of cerebrospinal fluid was not recognized within three punctures was in 2 cases (4%) in group A and in 1 case (2%) in group W. In these 3 cases, spinal anesthesia was performed easily using 25-gauge Whitacre. In group A, the spinal needle could be inserted without using an introducer in 35 cases (70%). The incidence of the postoperative headache or back pain was low and postdural puncture headache (PDPH) did not occur in both groups. We conclude that 26-gauge Atraucan can be handled easily and useful for preventing PDPH. PMID- 9411037 TI - [Anesthetic management for bronchoscopy in patients with congenital tracheomalacia]. AB - We experienced the anesthetic management for two patients with congenital tracheomalacia. Inhaled anesthetics are considered to worsen the respiratory condition of tracheomalacia because of its bronchodilating effect. Thus we tried awake intubation in one case, but it was difficult. In another case, we used slow induction with sevoflurane and the trachea was intubated smoothly. Inhaled anesthetics have possibility of worsening the degree of tracheomalacia and have been used very carefully. However, we considered that slow induction with inhaled anesthetics in children with congenital tracheomalacia is a safe and necessary technique, under careful observation of respiratory conditions. PMID- 9411038 TI - [Types and methods of identification of atypical antibodies posing clinical problems]. PMID- 9411039 TI - [Reflection on the progress in the study of tuberculosis in the 20th century]. PMID- 9411040 TI - [New concepts of corticosteroid treatment in pulmonary diseases of the premature infant]. AB - Corticosteroids for treatment or prevention of pulmonary disease are milestones in the management of preterm infants nowadays. Prenatal maternal administration as well as postnatal treatment of the mechanically ventilated preterm infant with bronchopulmonary dysplasia (BPD) are considered to be established concepts. However, the side effects of systemically given corticosteroids can be considerable (e.g., increased mortality, which cannot be explained by a specific side effect). In order to avoid as many side effects as possible and to attain an optimal response determining the best regimen is under current investigation. This review describes new concepts of corticosteroid application from the last years which have focused on "pulse"-therapy, inhalative therapy and early administration of corticosteroids. In conclusion, the trend is using corticosteroids as a prophylaxis for BPD in preterm infants instead of as a consequent treatment. PMID- 9411041 TI - [Measuring vibrations of transport stress in premature and newborn infants during incubator transport]. AB - BACKGROUND: Despite increasing numbers of centers for perinatology, transportation of newborns and premature infants can not totally be avoided for several obvious reasons. The following investigations were carried out as part of our quality control measures of the established neonatal transportation system, and were aimed on the optimization of a new neonatal transportation equipment resulting in reduction of transportation stress caused by acceleration forces. METHOD: The new system investigated consisted of a Volkswagen type T4 equipped with a Drager incubator type 5400, which was mounted on a pneumatic patient lift. We measured acceleration forces in three axes (expressed as K-Wert) as well as over a spectrum of frequencies (1-80 Hz). Measurements were taken at different points of the transportation unit during simulated transports driving a predetermined route. After obtaining informed consent of the parents, one actual transport of a newborn was used for an additional point measurement at the newborn's head. RESULTS: The mean K-Wert was decreased by about 50% in the vertical axis between the chassis of the car and the incubator by activating the pneumatic patient lift. Without activating the lift the K-Wert increased by about 20% between the car's chassis and the incubator. The frequency analysis showed resonance effects between the different components of the system. However, by activating the patient lift, effective accelerations in the incubator were decreased to less than 0.1 m/s2 across the whole frequency spectrum evaluated. The single measurement at a newborn's head revealed similar acceleration forces at the head of the baby and under its head. CONCLUSIONS: Utilization of a pneumatic patient lift can reduce acceleration forces. However, our results show that each system (car, incubator, and its base) has to be investigated and optimized for this purpose as a unit. Optimization of the complete system is necessary not only before its primary use but also in regular intervals over the years. Sometimes, further improvements can be reached with minor modifications such as the exchange of worn out rubber buffers. PMID- 9411042 TI - [Acute and chronic fetofetal transfusion syndrome]. AB - BACKGROUND: In the fetofetal transfusion syndrome we can discriminate between a chronic form with severe difference in weight and an acute form with a difference in haemoglobin between the twins. The aim of the retrospective study was to investigate the influence of both forms regarding infant morbidity and mortality. PATIENTS: Of the 135 twins treated from 1.1.1984 to 31.12.1993 in the Department of Pediatrics at the Christian-Albrechts-University Kiel, 27 (20%) showed evidence of fetofetal transfusion. RESULTS: Twelve (8.8%) fulfilled the criteria for an acute and 10 (7.4%) for a chronic transfusion. In 5 (3.9%) twins both forms were evident. Clear differences between twins with acute or chronic fetofetal transfusion syndrome could be seen in gestational age and birth weight. The twins with an acute fetofetal transfusion were significantly older (median: 31.5 vs. 29.0 weeks of gestation), and heavier (median: 1900 vs. 1020 g). Furthermore this group faired significantly better statistically when compared to the chronic form as regards the incidence of mortality (7 vs. 0), the Respiratory Distress Syndrome grades III and IV (7 vs. 2), persistent ductus arteriosus (13 vs 9), intraventricular haemorrhage (14 vs 6) and posthaemorrhagic hydrocephalus (4 vs 0). Similar significant differences could be seen as regards mortality, the Respiratory Distress Syndrome, and intraventricular haemorrhage when acute fetofetal transfusion was compared with the combined acute on chronic form. No significant difference regarding neonatal morbidity and mortality, however, could be shown between infants having undergone chronic fetofetal transfusion and infants with the combined acute on chronic form. CONCLUSIONS: It can be concluded that twins with chronic fetofetal transfusion show a higher rate of neonatal morbidity and mortality, whereas, the acute form does not significantly influence these factors. As both forms not only differ in onset of the disease, pathogenesis, and the actual course of the disease. It is important to always differentiate between the acute and chronic forms. PMID- 9411043 TI - [Pathogenesis, diagnosis, clinical and therapeutic aspects of ataxia telangiectasia]. AB - Ataxia-telangiectasia (AT) is an autosomal recessively inherited disease (one case in 40,000 to one case in 100,000 live births) whose principal features are oculocutaneous telangiectasia, progressive cerebellar ataxia, B- and T-cell immunodeficiency with recurrent sinopulmonary infections, sensitivity to ionizing radiation and cancer predisposition. The AT-gene (ATM) was recently identified by positional cloning on chromosome 11q22-23. In this paper the diagnostic, clinical and therapeutic problems of 9 AT-patients treated in our clinic are discussed in context with the current literature. Although all patients had discrete signs of cerebellar ataxia at infancy, there was a significant delay of definitive diagnosis (median 4, range 1.5-6.5). Elevated alpha fetoprotein levels clearly distinguish AT from other ataxias and immunodeficiency syndromes. PMID- 9411044 TI - [Conditioning of affect-induced breath-holding spells]. AB - Breath holding spells often arise in the context of affectively dramatic conflict situations between mother and child. Assessment by psychopathological screening instruments, however, has not given empirical evidence of an increased psychiatric morbidity in these children. Therefore, in our study we did not concentrate on basic psychopathology but on behavioral variables that might be effective during the ongoing attack episode and, hereby, exert an influence on the risk of chronification (relapse rate). The main goal of this approach is to examine secondary reinforcement effects on the attack behavior according to the learning principle of operant conditioning. Our sample consisted of 28 children and ten siblings as control group. To control for effects of behavioral disorders in the sample, we applied the Marburger Verhaltensliste (MVL) on the level of the child, and the Familienfragebogen (FFBO-III) on the level of family adaptation. The main assessment instrument, however, was the Functional Behavior Analysis (FBA) in order to measure the trigger, reaction and consequence conditions in the course of given attack episodes. MVL and FFBO-III results confirm the lack of basic psychopathology in the patients and their families. The individualized FBA's can be transformed in a taxonomy of five distinct types. All the first three types are triggered by intensive conflict situations and show a high relapse rate (type 1) if the mother reacts in a rewarding manner with positive consequences for the child (reinforcement condition), a dramatically reduced rate (type 2) if the mother reacts neutral (extinction condition), or a heterogeneous pattern (type 3) if the mother reacts punishing (punishment condition). In type 4 (pallid type) and type 5 (triggered spontaneously), respectively, no responsiveness to conditioning effects can be recognized. With respect to parent counselling, a recommendation for a quiet and consequent reaction can be concluded, especially in the case of a preceding conflict situation. The empirical results are integrated into a hypothetical model on pathogenesis that delineates the interaction of neurophysiological and behavioral factors in the maintenance of breath-holding spells. PMID- 9411045 TI - [Amyloidosis: classification, clinical characteristics, diagnosis and treatment]. PMID- 9411046 TI - [Biologically active food additives and their use in internal medicine]. PMID- 9411047 TI - [A long experience in basic therapy of rheumatoid arthritis]. AB - 183 patients with rheumatoid arthritis (RA) received basic drugs for 3 to 15 years. The patients were followed up for 13.5 years, on the average, with intervals 6-48 months. Clinical, laboratory, instrumental and morphological methods of investigation were used. The findings are reviewed on the mortality and causes of death, lethal tumors, overall complications and those observed in 55 patients on glucocorticoids. PMID- 9411048 TI - [The clinical role of C-reactive protein in rheumatoid arthritis]. PMID- 9411049 TI - [A comprehensive assessment of cardiac conduction system in hypertensive subjects]. PMID- 9411050 TI - [Time-dependent effect of metoprolol in middle-aged and old patients with angina]. AB - As shown by X-ray clinical and laboratory examinations of 495 patients over 18 years of age with primary tuberculosis of the respiratory organs, among primary forms of the specific process most prevalent are tuberculosis of the intrathoracic lymph nodes (tumor form), infiltrative tuberculosis with preroot or lower lobe location and exudative pleurisy. The peak of the incidence was reported in the group of 20-25-year-olds. The disease took an cute course with severe intoxication and symptoms of bronchopulmonary involvement in 45.3%, a torpid course with scare symptoms in 49.7% and a chronic course of primary tuberculosis in 5.0% of the examinees. Paraspecific reactions were observed in 5.9%, hyperergic tuberculin tests typical for primary tuberculosis in 10.9% of the cases. The examination of the sputum for bacterial and altered forms of M. tuberculosis raised frequency of the diagnosis verification by 16.0%. Respiratory tuberculosis in adults runs in the presence of marked disorders in lymphocyte T- and B-systems, antituberculous immunity and metabolic processes. Incidence rate and intensity of the above disorders correlate with severity of tuberculous intoxication and tuberculous lesions. The informative value of the complex of clinicoroentgenological and immunological tests in the diagnosis of primary tuberculosis varies from 44.4 to 79.6% and from 28.3 to 77.5% in dependence on the immunological and biochemical test, respectively. PMID- 9411051 TI - [Clinical features and diagnosis of primary tuberculosis of the respiratory organs in adults]. AB - 35 anginal patients with the disease functional class II-III were treated with metoprolol given in conventional regimen (100 mg twice a day) and adjusted to chronosensitivity to metoprolol (100 mg in a single dose). 23 patients were aged 50-72 years. Chronosensitivity was determined at acute pharmacological tests with control of blood pressure, stroke volume. It was found that the highest chronosensitivity to metoprolol in middle-aged and elderly anginal patients occurs with maximum for the stroke volume index in 15.32 and for ejection in 21.04. PMID- 9411052 TI - [Immunoglobulin G monoclonal human anti-rhesus Rho(D) to prevent rhesus incompatibility]. AB - RhD immunisation which follows pregnancy can be prevented by administration to the rhesus-negative mother of 20 micrograms anti-D immunoglobulin per 1 ml of D positive fetal red cells in the maternal circulation. With the aim of substitution of polyclonal anti-D with monoclonal one we developed human-mouse cell lines producing anti-RhD IgG1 by fusing EBV-transformed human immune lymphocytes with murine myeloma. After several clonings and passages the EBV genome was eliminated from the cell lines. The antibodies were purified from culture supernatants using protein A affinity chromatography and tested for sterility, virus contamination, pyrogenecity, toxicity and DNA content. The monoclonals were compared with the standard polyclonal anti-RhD in in vitro and in vivo assays. In antibody-dependent cell cytotoxicity (ADCC) 2 of 4 studied monoclonals promoted greater RBS lysis than polyclonal anti-D at equivalent concentrations. Detection of binding site number of monoclonals anti-D revealed about 10,000/RBS D-determinants on DCe/dce erythrocyte which agrees with the data for polyclonal anti-D. Best in ADCC monoclonal anti-D sharply increased human autologous 51Cr-labelled rbc sensitised in vitro as well as accelerated clearance of RhD RBS from circulation of Rh-negative volunteers injected with 150 micrograms monoclonal anti-D. After clinical trial using of monoclonal anti-D is permitted in Russia. PMID- 9411053 TI - [Disorders of regulation]. PMID- 9411054 TI - [Beta-carotenoids and their potential use in diseases of the digestive tract]. AB - Flying personal with chronic hepatitis received beta-caratinoid vetoron. The latter proved to be an effective immunomodulator improving lipid peroxidation, clinical and morphological characteristics in the personnel with chronic persistent hepatitis. Follow-up gives proves on longer physical fitness for service of those pilots who have taken beta-caratinoids. PMID- 9411055 TI - [Hyperglycemia in patients with purulent conditions]. AB - Basing on their rich clinical and experimental experience the authors conclude that hyperglycemia in purulent conditions results from receptors insufficiency of somatic cells membranes which arises due to either their block or destruction by endotoxicosis factors which are likely to be of leukocytic nature. Early cleansing of the purulent focus and detoxication provide normalization of the carbohydrate metabolism. PMID- 9411057 TI - [Effects of deep reflex-muscular massage and exercise on regulatory processes in the body]. PMID- 9411056 TI - [Clinical manifestations and treatment of intestinal dysbacteriosis in patients with Flexner's dysentery]. AB - Flexner's dysentery is often accompanied with intestinal dysbacteriosis. Disbiotic changes in the intestine contribute to specific shigella endotoxins entering blood flow thus prolonging clinical symptoms of the underlying disease. Administration of bacterial biological preparations (bifidumbacterin forte, lactobacterin) relieves specific endotoxemia, reduces the duration of the disease and has an immunomodulating action. PMID- 9411058 TI - [A new prokinetic drug cisapride: pharmacological properties and clinical potential]. PMID- 9411059 TI - [Our experience with acycloguanosine treatment of herpes simplex]. AB - A 5% ointment acycloguanosine was tried in the treatment of recurrent herpes simplex of the skin and genitalia in 48 patients. The treatment brought limitation of the process, relief of the symptoms, control of local manifestations and toxicity, accelerated epithelization, complete regression and disappearance of the eruption. PMID- 9411060 TI - [Factors responsible for the course and long-term outcomes of bronchial asthma: chemotherapy]. AB - In a 10-year retrospective observational study the authors investigated the effects of antiinflammatory therapy with corticosteroids (CS), broncholytic therapy with theophylline drugs (TD) and of inhalations of short-action sympathomimetics (ISM) on the course and long-terms outcomes of bronchial asthma (BA). The data were processed using actuarial analysis. It was established that long-term CS therapy diminishes the risk of the unfavourable BA outcome (death, life-threatening conditions, invalidism). The protective CS action was maximal in patients with severe persistent BA. The best treatment results were achieved in long-term CS course in doses warranting effective control over respiratory symptoms. Long-term TD administration failed to demonstrate a significant influence on long-term BA outcomes. Overdoses of ISM occasionally observed in the course of relevant treatment placed the exposed patients at higher risk of unfavourable BA outcome. ISM overdoses were associated with severe and uncontrollable BA. PMID- 9411061 TI - [Infectious endocarditis with negative hemoculture: diagnosis and treatment]. PMID- 9411062 TI - [Anemia]. PMID- 9411063 TI - [Diarrhea in differential diagnosis of infectious diseases (dedicated to the 100th anniversary of the birth of the outstanding clinician, academician of the Russian Academy of Medical Sciences, outstanding scientist of Russian Federation Alexander Fedorovich Bilibin)]. PMID- 9411064 TI - [Baby blues and hospital rooming-in of mother and child. Mourning the myth of the ideal mother]. PMID- 9411065 TI - Shaping the revolution: thoracic surgeons and something more. Presidential address. PMID- 9411066 TI - [Participation of various bacterial species in translocation in intestines of rats subjected to stress]. AB - In the rats, which were treated with impairing factors such as: malnutrition, cooling down and ischaemia, translocation was studied. Aerobes as well as anaerobes which penetrated beyond the intestine into the mesenterium, liver, spleen and kidneys were sought. Bacteria in peripheral blood were not found. The results show that the type of bacterium undergoing translocation does not depend on the factor impairing "intestinal barrier" and translocation mechanisms are most probably the same for all the bacteria found. PMID- 9411067 TI - [Lysogenic conversion as a factor influencing tolerance to vancomycin in Staphylococcus aureus]. AB - The standard, non-lysogenic, bacteriophage-free S. aureus NCTC 8325-4 strain was lysogenized with 15 different, obtained in our laboratory staphylokinase converting bacteriophages belonging to serological groups A, B and F. MIC and MBC of vancomycin as well as the ratio of MBC to MIC were evaluated for all 15 lysogenic derivatives. The obtained results were compared with those for maternal strain. In the case of eight strains the ratio MBC/MIC showed the presence of tolerance to vancomycin (MBC/MIC > or = 32). Four of the vancomycin-tolerant derivatives were lysogenized with bacteriophages belonging to the serological group A, two were members of group B and two belonged to group F. PMID- 9411068 TI - [Comparison of several methods for detection of methicillin resistance in clinical strains of Staphylococcus sp]. AB - 108 Staphylococcus spp. strains from 300 clinical specimens from hospitalized patients were isolated. Identification and drug resistance were determined using automated ATB system. 37 S. aureus strains, 44 S. epidermides strains and 27 strains of other coagulase-negative staphylococci were cultured. Sensitivity to methicillin of S. aureus was determined with four methods: ATB system, disc diffusion (Oxa 1 microgram), Crystal MRSA ID System and agar screen test in TSA medium with methicillin (25 micrograms/ml). 13 S. aureus strains (about 1/3 of strains) were methicillin-resistant (MRSA). Complete conformity of the results was obtained with Crystal MRSA ID, disc-diffusion and agar screen tests. In the case of three S. aureus strains the results of determination in ATB system were not consistent with the results obtained with the use of the methods mentioned above. Susceptibility to methicillin of 71 coagulase-negative strains (CNS) was determined using two methods at first: ATB and disc-diffusion. In the case of 25 methicillin-resistant strains identical results were obtained. For 20 coagulase negative strains non-conformity with the results of these two methods was observed. As the decisive method, the agar screen test (TSA-MET) was applied. 18 of these 20 CNS strains were categorized as methicillin-resistant. Finally, 43 MRCNS (i.e. 60%) were detected among 71 coagulase-negative strains. The results of methicillin resistance determination of staphylococci in an automated system should be confirmed with a second test such as agar screen, disc-diffusion or Crystal MRSA ID System (in the case of S. aureus). PMID- 9411069 TI - [Incidence of methicillin-resistant and methicillin-susceptible Staphylococcus strains in clinical materials]. AB - 365 S. aureus and 165 CNS were evaluated for susceptibility to methicillin. They were received from various clinic of Clinical Hospital in Bydgoszcz. The bacterial cells that grew in the presence of 25 mg methicillin/L were evaluated as methicillin-resistant. It was found that the 129 MRSA and MRCNS. Incidence depended on the kind of clinic and material. Most often they were isolated from clinic: Surgery, Neurosurgery, Intensive Care Unit, Neurology and Ortopedics. They were from: intubation tube, catheters and vascular grafts. The methicillin resistant S. epidermidis and S. haemolyticus showed above 50% resistance to methicillin. PMID- 9411070 TI - [Susceptibility to cefazolin, cefamandole and ceftazidime in Staphylococcus strains]. AB - 365 S. aureus and 165 Coagulase Negative Staphylococci were tested for susceptibility to methicillin, cefazolin, cefamandole and ceftazidime by standard broth microdilution and disc method. In staphylococci resistance to methicillin normally parallels resistance to beta-lactams, and it has been suggested that cephalosporins are not be used clinically if susceptbility tests show resistance to methicillin. Populations of methicillin resistant staphylococci are made up of a mixture of methicillin-susceptible and methicillin-resistant cells (heteroresistant isolates). This phenomenon is the cause of differences in resistance in vitro to methicillin and tested cephalosporins. Cefamandole was the cephalosporin which retained most antibacterial activity against some methicillin resistant isolates (MICm MRSA = 63.2 mg/L, MICm MRCNS = 67.4 mg/L). This antibiotic has extremely small resistant subpopulations, only detectable by high inoculum screening or prolonged incubation of isolates in the presence of the drug. Cefazolin was the cephalosporin which had most antibacterial activity against all methicillin-susceptible isolates (MICm MSSA = 1.3 mg/L, MICm MSCNS = 1.6 mg/L). Among the 3 studied cephalosporins ceftazidime was found to be the least active against methicillin-resistant and methicillin-susceptible Staphylococci (MICm MSSA = 25 mg/L, MICm MRSA = 334.5 mg/L, MICm MRCNS = 414.6 mg/L, MICm MSCNS = 26.7 mg/L). PMID- 9411071 TI - [Animal systemic iron sources utilized in vitro by staphylococci]. AB - Under iron-restricted conditions staphylococcal strains could utilize in vitro several animals body iron sources in form of bovine haemoglobin, hemin, lactoferrin and transferrin, ovotransferrin, horse myoglobin ferritin and cytochrome C. Spectrum of utilized iron sources was not dependent on species affiliation and kind of siderophores system. Strains isolated from clinical materials utilized largest spectrum of animal iron body sources. PMID- 9411072 TI - [Selected properties of Staphylococcus aureus strains using phenotype to show a lack of coagulase synthesis]. AB - Eighteen of S. aureus strains isolated from clinical specimens and showing negative reaction for coagulase in conventional tube test were characterized by using five different methods: ID32 Staph, API Staph, Gpl-15, STAPH-ZYM (Rosco) and classical tube methods. The membership of all examined strains to S. aureus subsp. aureus was confirmed. None of the strains produced detectable amounts of coagulase but all of them produced protein A and clumping factor. All of them were typable with phages of the III lytic group and 11 of them possessed phage pattern characteristic of the 83A complex. All the investigated hospital strains were resistant to methicillin. Five showed homogenous and 13 heterogenous type of methicillin resistance. Moreover, all of them were resistant to tetracycline, gentamicin, tobramycin and kanamycin and to MLS group of antibiotics. PMID- 9411073 TI - [Occurrence of Moraxella catarrhalis in patients with respiratory tract infections]. AB - The study was undertaken to evaluate the incidence of Moraxella catarrhalis in patients with respiratory tract infections. Overall 514 specimens including 370 throat swabs and 114 sputum specimens were examined. The 78 strains isolated basing on morphological and biochemical characteristics were classified as Moraxella catarrhalis. The sensitivity of the strains to antibiotics was also estimated. The frequency of M. catarrhalis isolation from the throat swabs (15.9%) was higher than from the sputum (13.2%). Selected 25 specimens of sputum were tested simultaneously by quantitative and qualitative methods. Quantitative method was more sensitive (84% positive findings) than qualitative method (60% positive findings). Resistance to ampicillin was found in 52 (66.7%) strains of M. catarrhalis determined mainly by beta-lactamase production (over 70% strains were producers of beta-lactamase). All strains were sensitive to ofloxacin and amoxycillin/clavulanic acid combination. The frequency of M. catarrhalis isolation was higher in autumn-winter period than in summer (May-September). We conclude that M. catarrhalis, beside Streptococcus pyogenes (20.2%) and Streptococcus pneumoniae (17.1%), are the most frequently isolated bacteria in patients with respiratory tract infections. PMID- 9411074 TI - [Detection of endotoxins and enterotoxins of Bacteroides fragilis in culture media]. AB - Four B. fragilis strains were studied one nonenterotoxigenic (NTBF) and three enterotoxigenic (ETBF). Endotoxin and enterotoxin which are released into the culture medium during the growth of strains were detected. Cultures in BHI broth were incubated for 48 hours at 37 degrees C. After 4, 8, 16, 24 and 48 hours of cultivation, samples of bacterial culture were collected and the optical density was measured. Then the samples were centrifuged, supernatants were filtered through 0.45 micron filters and concentrated three times with 5000 D ultrafilters. Prepared samples were kept frozen at 70 degrees C until use. The presence of endotoxin in samples was revealed by means of immunoelectroprecipitation (IEP) and immunoenzymatic test (dot-ELISA). The assays were performed with antibacterial rabbit immune sera. The activity of enterotoxin was detected on a human colon adenocarcinoma cell line HT 29/C1. The results of the study indicate that endotoxin is released spontaneously by nonenterotoxigenic IPL E 323 strain into the culture medium at the early stages of cultivation. The presence of endotoxin is not demonstrated by means of immunoelectroprecipitation in culture filtrated of ETBF strains. Trace amounts of endotoxin are revealed with dot-ELISA. The activity of enterotoxin is detected after 16 hours of incubation of ETBF strains. PMID- 9411075 TI - [Evaluation of the reliability of serological diagnostic tests for Mycoplasma pneumoniae infection carried out by laboratories in sanitary epidemiological stations (WSSE)]. AB - In the action undertaken for evaluating of the reliability of serological tests for M. pneumoniae infection 33 laboratories of the Sanitary Epidemiological Stations participated. Every laboratory had to determine twice at an interval of 2-4 weeks the level of mycoplasma antibodies by the complement fixation test in serum samples divided into 4 groups: sera not containing these antibodies-titre < 5, or containing them in titres of 60, 120 and 320. The correct results of the determinations were obtained in 27 laboratories (81.8%) for samples without M. pneumoniae antibodies, and in 22 (66.6%) and 14 (42.4%) for samples with titres of 60 and 120, and 320 respectively. Only 4 laboratories (12.1%) obtained correct results of these determinations for every sample and in both testing series. In these series considered separately correct results were obtained in 9 (27.3%) and 8 (24.2%) laboratories. Faulty results in all samples in both testing series were reported from 2 (6.1%) laboratories. In the individual series all false results were obtained in 4 (12.1%) and 3 (9.1%) laboratories. The study showed that for raising of the quality and reliability level of serological investigations for M. pneumoniae infection a permanent practice should be periodic training of laboratory workers and frequently repeated interlaboratory controls of the reliability of test results. PMID- 9411076 TI - [Analysis of aerobic and anaerobic bacterial flora colonizing drains after surgical abdominal incisions]. AB - Aerobic and anaerobic bacterial flora of post-operative incisions with drainage were examined. From each of 28 patients three specimens were taken; during operation (smear from peritoneal cavity), liquid from drain (taken at 3-th day after operation) and smear from drain taken at the end of drainage. Enterococci, Enterobacteriaceae spp. and anaerobes, especially Bacteroides spp. were most often isolated from specimens taken during operation. Enterococci and coagulase negative Staphylococci-often resistant to methicillin, were most often isolated from specimens taken at the end of drainage. PMID- 9411077 TI - [Bacterial flora in chronic inner ear infections in adults]. AB - The aim of the study was to analyse microbiologically middle ear exudate obtained from 56 patients, aged 17 to 83 years, treated for chronic otitis media. Aerobic bacteria only were found 49 patients (87,5%). Mixed aerobic and anaerobic isolates were recovered from 7 patients (12,5%). The most common bacteria isolated from the middle ear exudate, in descending order frequency, were Staphylococcus aureus (45%), Pseudomonas aeruginosa (34%), Proteus mirabilis (16%) and Prevotella melaninogenica (9%). Other organisms were isolated less frequently. In 34 patients only one isolate was recovered, in 22 patients the isolated bacteria coexisted with other microorganisms. PMID- 9411078 TI - [Aerobic and anaerobic bacterial flora in chronic sinusitis in adults]. AB - The aim of the study was to analyse microbiologically samples obtained from 30 patients aged from 21 to 73 years treated for chronic sinusitis. Aerobic bacteria only were isolated in 16 patients (53%), and anaerobic organisms only in 5 patients (17%). Mixed aerobic and anaerobic isolates were recovered from 9 patients (30%). The isolated aerobic bacteria were as follows: streptococci from the species Streptococcus salivarius, Streptococcus anginosus, Streptococcus group C, Streptococcus sanguis, Staphylococcus aureus, Gram-negative rods from the genus Haemophilus and rods from the Enterobacteriaceae family, and strains of Moraxella catarrhalis. The isolated anaerobic microorganisms Gram-negative rods from the genus Prevotella, Bacteroides, Fusobacterium, Gram-positive cocci from the genus Peptostreptococcus. Other organisms from the genus Vailonella, Eubacterium and Actinomyces were isolated less frequently. In 15 patients only one isolate was recovered, in 15 patients isolated bacteria were mixed with other microorganisms. PMID- 9411079 TI - [Microflora of periodontal pockets in advanced periodontitis]. AB - The aim of study was the evaluation of periodontal pockets microflora in patients with advanced periodontitis. From each subject 16-20 samples were taken using paper points. Pooled sample after 60 s. mixing was serially diluted in reduced BHI. For total cell counts and for the isolation of black pigmented anaerobes Brucella agar supplemented with 5% sheep blood, hemin, menadione, with and without Kanamycin-Vancomycin mixture and BM agar plates were used. For isolation of A. actinomycetemcomitans TSBV agar plates were used. Cultures were incubated in anaerobic chamber at 37 degrees C for 7 days and TSBV agar plates in an atmosphere of 95% air-5% CO2 at 37 degrees C for 5 days. Microorganisms were identified by Gram staining, colony morphology, fluorescence in UV-light, haemagglutination of 3% sheep erythrocytes, fermentation of sugars, production of indole, urease (API 20A), specific enzymes (Rapid ID 32A). Twenty seven subjects with clinically recognized periodontitis were examined. Microorganisms important in periodontitis were isolated from periodontal pockets of almost all examined subjects. The number of bacteria obtained from the sample of one patient ranged from 1 x 10(4) CFU/ml to 3,6 x 10(6) CFU/ml. Porphyromonas gingivalis was identified in the samples taken from 17 patients, Prevotella intermedia-19, Actinobacillus actinomycetemcomitans -11, Fusobacterium nucleatum-9, Peptostreptococcus spp.-22. PMID- 9411080 TI - [A new book is the result of research on Ivan Bratt by HAkan Westling. "Bratt did not want rationing"]. PMID- 9411081 TI - [Research scientists, cheats or simply weirdos? The borders are floating in a complicated scientific world]. PMID- 9411082 TI - [Political control of the universities is contra-productive and hazardous. Free universities are important for democracy]. PMID- 9411083 TI - [Medical opinions make nobody healthier]. PMID- 9411084 TI - [What does a gene care about prohibition? A promiscuous gene flow between bacteria is a growing problem]. PMID- 9411085 TI - [Does the infectious disease, human ehrlichiosis, exist in Sweden? Ticks get a hold of new zoonoses]. AB - Human ehrlichiosis diseases, decently recognised as emerging human infections in the USA, are caused by vector-borne, strictly intracellular bacteria of the family Rickettsiaceae. Human monocytic ehrlichiosis is caused by Ehrlichia schaffeensis, whereas the agent causing human granulocytic ehrlichiosis (HGE) has yet to be identified [1]. The putative increase in the occurrence of these primary zoonoses is dependent on the complex relationship between the infectious agents, the vectors, and the hosts (rodents, deer) which constitute the wild-life reservoir. In Scandinavia, granulocytic ehrlichiosis is well known in veterinary medicine. Pasture fever in cattle and sheep is caused by Ehrlichia phagocytophila, whereas granulocytic ehrlichiosis in horses and dogs is caused by a new, recently characterised Ehrlichia species [2]. All species of Ehrlichia causing granulocytic ehrlichiosis are closely related both genetically and antigenically, and are all transmitted by ticks of the genus Ixodes. In Sweden, veterinary cases of granulocytic ehrlichiosis are characterised by a geographical distribution corresponding well with that of Ixodes ricinus, and a seasonal distribution similar to that of Lyme borreliosis. In Europe, clinical cases of HGE have so far been reported only from Slovenia [5], though seroprevalence figures of 8-17 per cent have been reported for tick-exposed populations [6, 7]. As most cases are probably subclinical, and as clinical symptoms, when present, are non-specific, clinical diagnosis is dependent on the clinician's awareness of the existence of the disease. Laboratory diagnostic tests are now available at Kalmar. PMID- 9411086 TI - [Care of a patient with a rare and highly contagious virus disease. An emergency situation resulted in good preparedness]. AB - Ever since the eradication of smallpox, Sweden has been poorly furnished with emergency facilities for the care of patients with serious, very infectious diseases. National interest in creating such facilities was aroused by epidemics of haemorrhagic disease (first and foremost due to Ebola virus during the present decade), at the same time as the first Scandinavian case of haemorrhagic fever associated with a risk of person-to-person infection occurred in Linkoping. A special laboratory which has been set up at the Centre for Disease Control, in Stockholm, and University Hospital, Linkoping, in collaboration with the Board of Health and Welfare, has introduced a high-security infectious disease unit for the care of such patients, with separate ventilation and waste-water treatment systems. The unit is also equipped to provide intensive care, and a laboratory can be rapidly set up and fully operative within 12-24 hours. Most important of all, personnel are available who are trained both for laboratory work and the care of such patients, and used to working as a team and familiar with the special protective equipment. If a patient can not be transported to the special unit, a team is available to travel to the hospital where the patient has been admitted, to give instruction and help to set up infection control routines and even supply protective equipment. PMID- 9411087 TI - [Animal husbandry and food handling today. Current food preservation has changed the intestinal flora]. AB - In the modern world, as refrigerated storage has replaced lactic acid fermentation methods of food preservation, we no longer ingest large quantities of live lactobacilli with our food, but relatively large numbers of potentially pathogenic Gram-negative bacteria instead. In the brave new world, antibiotics are used as growth-promoting additives in animal feed in every country but Sweden. The antibiotics used in Europe at low dosages, and exempt from prescription requirements, include macrolides, streptogramins and nitro imidazoles. There is reason to believe that avoparcin, recently banned in Europe as an antibiotic animal feed additive, has contributed to the occurrence of vancomycin-resistant enterococci in humans. Animal foodstuffs production in Europe and the USA is so intensive that it provides very favourable conditions for the spread of bacteria. Pig, cattle and poultry breeding in Europe are beset by formidable difficulties in the form of Salmonella infections, which constitute not only a hygiene but also a clinical problem. Sweden, the bravest of brave new worlds, faces an increase in the influx of domestic animals and foodstuffs from other countries, and in general less restrictive regulations concerning food production and animal husbandry, but reduced powers of control. Thus, we may expect problems both with pathogens (traditional ones such as Salmonella, and potential ones such as food-spoilage Gram-negative bacteria), and with antibiotic resistant bacteria in our food-stuffs, to become greater. PMID- 9411088 TI - [Gynecologic laparoscopy. Advantages or disadvantages, which is more important?]. PMID- 9411089 TI - [Scrutinizing colleagues assure quality]. PMID- 9411090 TI - [Anxiety worse than pain among patients waiting for heart surgery]. PMID- 9411091 TI - [A new surgical technique in atrial fibrillation. The maze procedure restores sinus rhythm]. AB - Atrial fibrillation (AF) is a common arrhythmia associated with significant morbidity and increased mortality, partly due to the increased risk of stroke. The maze procedure, introduced by James Cox of the USA, is an internationally established surgical alternative in cases of unsuccessful medical or catheterised treatment of paroxysmal or chronic AF. It is an open heart procedure, involving multiple transmural incisions and continuous suture lines in both atria. By creating a maze of atrial tissue, the re-entrant circuits causing the AF are interrupted, hence re-establishing regular sinus rhythm and atrioventricular synchronization. The article reviews the initial 3-year experience of the procedure in 10 patients with AF, either paroxysmal (n = 5) or chronic (n = 5). The indications for surgery were disabling symptoms in all 10 cases, medical treatment failure in nine cases, previous AF-associated stroke in three cases, and a significant atrial septal defect in one case. All patients underwent extensive investigation both pre- and post-operatively. Postoperatively, nine of the 10 patients manifested regular sinus or atrial rhythm and freedom from or amelioration of preoperative symptoms associated with AF. There were no deaths, neurological complications or long-term recurrence of arrhythmia. One patient had an early recurrence of AF that was not amenable to medical treatment, and was subsequently treated with His' bundle ablation. Of the remaining nine patients, seven manifested signs of some postoperative atrial contraction at echocardiography, the occurrence of which needs to be borne in mind with a view to reducing the risk of future thromboembolic events. We recommend the maze procedure as an attractive surgical option in cases of unsuccessful medical treatment of paroxysmal or chronic AF. PMID- 9411092 TI - [Hepatitis GBV isolated in acute liver failure. Successful outcome after liver transplantation]. AB - Fulminant hepatic failure is a life-threatening condition associated with a mortality of approximately 80 per cent. Liver transplantation may be the only life-saving recourse in such cases. The condition can be caused by any of a number of different agencies such as viral infection, or toxic, circulatory or metabolic factors, though in a large proportion of cases the aetiology is unknown. Recently, knowledge has accumulated of a new hepatitis virus, hepatitis GB virus (HGBV), a Flavivirus remotely related to hepatitis C. The clinical significance of this virus is unclear. It is found in 3-4 per cent of blood donors, and most HGBV-positive patients are asymptomatic though some develop fulminant hepatic failure. The article consists in a case report of fulminant hepatic failure in a 17-year-old woman where no possible aetiological factor could be identified, other than her HGBV-positivity. The patient underwent a successful liver transplantation and is now, 18 months later, in excellent condition. She is still HGBV-positive but manifests no hepatic effects. Whether HGBV infection was responsible for the hepatic failure remains unclear, however. PMID- 9411093 TI - [Large arterial thrombosis. Rapid recanalization with a new thrombocyte inhibitor]. AB - A 56-year-old male cigarette smoker, without other risk factors for arterial disease and no history of heart disease or thrombotic events, was admitted as an emergency case because of chest pain shortly after a fall. He had a right coronary artery occlusion, which was managed with angioplasty and stenting. A large thrombotic occlusion in the distal aorta probably occurred simultaneously with the coronary occlusion, became symptomatically manifest and was diagnosed when the chest pain subsided after stenting. This thrombus was successfully treated with the new platelet inhibitor abciximab. PMID- 9411094 TI - [Report of a case. Inhalation steroids caused adrenal cortex suppression]. AB - In one of two cases of systemic effects of high-dose inhaled corticosteroid treatment with fluticasone propionate, adrenal suppression was demonstrated in a young man, and in the other retarded growth and severe general effects were observed in connection with infection in a child. These cases illustrate the risk of systemic effects of inhaled corticosteroids, and the importance of using the lowest possible maintenance dose. PMID- 9411095 TI - [Pharmacists in hospitals optimize treatment. Pharmacology in practice for physicians and patients]. PMID- 9411096 TI - [Goran Tunstrom about his experiences as a patient. "The round feels humiliating"]. PMID- 9411097 TI - [Involuntary sterilizations: the medical community risks to be in the dock!]. PMID- 9411098 TI - ["The house doctor" Gerard Valance concentrates on home visits. A French physician is always (almost) willing to help]. PMID- 9411099 TI - [Professor Anne-Liis von Knorring: hyperactive children as a risk group for future antisocial development]. PMID- 9411100 TI - [How to handle the question of screening in diabetes?]. PMID- 9411101 TI - [Respect your signature]. PMID- 9411102 TI - [Enthusiasm for ultrasound creates problem for the examining physician]. PMID- 9411103 TI - [Indirect tobacco advertising in the Lakartidningen!]. PMID- 9411104 TI - [Centralizing neonatal intensive care!]. PMID- 9411105 TI - [Concentrate more on trauma research! Injuries have enormous social and economical consequences]. PMID- 9411106 TI - [Dim the light! Glare problems in a world with ever increasing demands on visual acuity]. AB - Ocular tissue transparency is dependent on the regular lattice configuration of lens and corneal fibres of uniform diameter. Ageing is associated with degeneration of both lens and cornea, which lose some of their structural order and eventually their transparency, though this process is not uniform. The structural changes are local and result in ocular media opacities. When the opacities increase in number and extension, they begin to affect visual acuity. As the loss of acuity becomes clinically significant, we speak of clinically relevant cataract. Retinal exposure to light diffused by intraocular light scattering induces optical glare, one of two forms of glare. The other form is transient glare--i.e., glare due to adaptation problems in an environment with rapidly changing ambient luminance. Contemporary society is characterised by increasing emphasis on visual information in such forms as texts, icons, signs and symbols. The computer revolution has been accompanied by further stress on the importance of the detection and interpretation of written instructions. PMID- 9411107 TI - [Unethical research in developing countries is supported by western governments]. PMID- 9411108 TI - [Molecular mycobacterium-diagnostics. Weapon against increased global spread of tuberculosis]. PMID- 9411109 TI - [Suspect diaphragmatic injury in abdominal/thoracic trauma]. PMID- 9411110 TI - [Trauma education for nurses in Sweden. An active engagement for life-saving care]. PMID- 9411111 TI - [Patients write about the time before and after trauma. Life changed in a second....]. PMID- 9411112 TI - [Maria Magdalene Rudbeck--"Leonardo of the mentally sick" hospitalized for 53 years]. PMID- 9411113 TI - [ECG findings with poor prognosis. Bifascicular block as a warning sign in cases of syncope]. AB - Of 517 emergency room admissions for syncope at four Stockholm hospitals, 36 (7 per cent) were found to be cases of bifascicular block. Of these 36 patients, 34 underwent ambulatory electrocardiography (ECG) in hospital, which showed significant arrhythmia to be present in three cases [high-grade atrioventricular block (n = 2), or sustained ventricular tachycardia (n = 1)]. After a mean follow up of 32 months, eight more patients had developed high-grade atrioventricular block, and 13 patients had died (eight of them suddenly). It is concluded that patients with bisfascicular block constitute a high-risk category, and would benefit from thorough investigation, including an invasive electrophysiological study, in order to identify those at risk of progression to high-grade atrioventricular block or malignant ventricular arrhythmia. PMID- 9411114 TI - [Personal archives of Axel Hojer are now open for research. The controversial family physician of the Swedish Welfare State]. PMID- 9411116 TI - [Hans Wigzell on research and creativeness. "Thank God for a real argument"]. PMID- 9411115 TI - [Children in poor countries also have right to good health care. A new care program will reduce child mortality]. AB - According to the WHO (World Health Organization), 12 million children die annually before reaching the age of five. Seventy per cent of the deaths are related to one or more of five common diseases: acute respiratory tract infection, diarrhoea, measles, malaria and malnutrition. Consequently, drawing on international experience and expertise, the WHO and UNICEF (United Nations International Children's Emergency Fund) have compiled and developed guidelines for the primary care of Third World children. This programme, entitled Integrated management of childhood illness (IMCI), is expected to improve the care of children in areas and situations resources are limited. PMID- 9411117 TI - [Future primary health care--a prime community care?]. PMID- 9411118 TI - [He was rewarded with Nobel Prize for discovery of prions]. PMID- 9411119 TI - [Patients can feel safe after dermatologists' assessment]. PMID- 9411120 TI - [Early diagnosis is crucial for prognosis of skin melanoma]. PMID- 9411121 TI - [Oncology specialists are not competent to assess the work of a dermatologist]. PMID- 9411122 TI - [An error of the figures in the article on malignant skin melanomas]. PMID- 9411123 TI - [It must be possible to administer a prescribed drug]. PMID- 9411125 TI - [Increasing use of antimycotics. Important to monitor resistance development]. PMID- 9411124 TI - [New discoveries on adenosis and prostacyclin. Physiological processes are surveyed in genetic elimination trials]. PMID- 9411126 TI - [Difficult to diagnose head injuries in drunk and violent persons]. PMID- 9411127 TI - [Overview of knowledge of health effects of smokeless tobacco. Increased risk of cardiovascular diseases and mortality because of snuff]. AB - Oral snuff, used by 20% of all Swedish males in 1996, causes blood nicotine levels similar to those in smokers. In a report by the National Board of Health and Welfare in 1997, it is concluded that scientific evidence of oral snuff as a cause of oral cancer is weak. Epidemiological studies of more than 100,000 construction workers showed hypertension (blood pressure > 160/90) to be more common among oral snuff users than among non-users. Twelve-year follow-up showed the relative risk of death due to cardiovascular disease to be 2-fold greater among snuff users, but 3-fold greater among smokers, as compared to never-users of tobacco (p < 0.001). The risk of death due to cancer was similar among snuff users and among never-users. In a clinical study of 151 healthy middle-aged men, 24-hour monitoring showed daytime heart rates and blood pressure to be significantly higher in snuff users > or = 45 years of age than in age-matched non-users (p < 0.05). The estimated 10-year future risk of cardiovascular events was 13.2% for smokers (p < 0.001) and 4.6% for oral snuff users (p = 0.3) as compared with 3.4% for never-users. As compared with non-users, oral snuff users manifested neither significant signs of accelerated atherosclerosis nor significantly reduced maximum physical capacity. Oral snuff usage causes physiological nicotine dependence, and results in the release of sympatho adrenergic stimuli associated with an increased risk of cardiovascular stress, which might in turn exacerbate the risk of fatal cardiovascular events. However, oral snuff usage does not seem to be associated with the same risk of accelerated atherosclerosis as is smoking. PMID- 9411128 TI - [Mercury in medical history. From curative to dangerous effects]. PMID- 9411130 TI - [Bo Jordin, the National Board of Health and Welfare, on leadership problems in health care: physicians and administrators don't speak the same language]. PMID- 9411129 TI - [Beta-2-agonists are still of value. Tolerance does not affect the bronchodilating effect]. AB - Use of the potent bronchodilators, beta 2-adrenoceptor agonists, has been a cornerstone of the treatment of obstructive lung disease, especially asthma, for the past 30 years. However, the occurrence of side effects and the development of tolerance have been discussed as limitations to their use. beta 2-Adrenoceptors are located on the surface of most cell types throughout the human body, and treatment with beta 2-agonists may exert effects in a wide variety of tissues. As with other pharmacological receptors, beta 2-adrenoceptors in most tissues develop tolerance as a result of continuous beta 2-stimulation. However, the bronchodilatory effect appears to be unaffected by the development of tolerance. Interestingly, most studies have yielded evidence suggesting tolerance development regarding protection against bronchoconstrictor stimuli such as methacholine, adenosine and exercise. Although the protective effect of a beta 2 agonist becomes attenuated with continuous treatment, this tolerance is partial and adequate residual protective effect remains. PMID- 9411131 TI - [High frequency of breast feeding in spite of restructuring of maternal health centers]. PMID- 9411133 TI - [When manpower is complete...a dream in primary health care]. PMID- 9411132 TI - [Anniversary of surgical gloves. "100 years' anniversary of cooked hands]. PMID- 9411134 TI - [Protect the interns from arbitrary rules! Internship and residency plans are not to be changed without sound reasons]. PMID- 9411135 TI - [The career is more important than a husband and children]. PMID- 9411137 TI - [Extensive material forgotten in a study on sleep apnea]. PMID- 9411136 TI - [All private practice was forgotten when surveying sleep apnea therapy]. PMID- 9411139 TI - [The majority of ambulance interventions are in the field of internal medicine]. PMID- 9411138 TI - [Courses on personality development are questioned]. PMID- 9411140 TI - [DAMP and dyslexia--don't listen to the fashionable slogans]. PMID- 9411141 TI - [Patients with Creutzfeldt-Jakob syndrome are screened. So far 122 cases discovered in Sweden]. PMID- 9411142 TI - [How to avoid diagnostic errors?]. PMID- 9411143 TI - [Diverticulitis is increasing among the elderly. Significant cause of morbidity and mortality]. AB - The prevalence of diverticulosis in western countries has increased and two thirds of the population over the age of 85 are now affected. Diverticulitis results from inflammation and subsequent perforation of a colonic diverticulum. Mild forms of diverticulitis usually present with gradually increasing symptoms from the lower left quadrant of the abdomen, whereas acute complicated disease is characterised by dramatic onset of abdominal pain, followed by fever within a few hours. The standard treatment for uncomplicated diverticulitis is bowel rest, with liquid diet or intravenous fluids in combination with antibiotics. Patients not responding to conservative treatment within the first 24 hours require further evaluation by computed tomography or ultrasonography. If an abscess is present, it can often be drained percutaneously. In cases of perforation and peritonitis, surgical intervention is mandatory, though no consensus exists as to the choice of procedure. Fistula formation and intestinal obstruction are also indications for surgical intervention, although the frequent recurrent attacks which commonly afflict these patients are seldom associated with severe complications. Prophylactic resection is not to be recommended for patients with diverticular disease, but a high-fibre diet may afford protection by preventing further complications. PMID- 9411144 TI - [Goya's diphtheria was in reality a sausage exchanged for a turnip]. PMID- 9411145 TI - [Let the stump be free in appendectomy]. PMID- 9411146 TI - [A compilation of "diagnostic errors" in Swedish health care. Missed diagnosis is most often a fracture]. PMID- 9411147 TI - [Better routines when handling arm fractures result in fewer diagnostic errors]. PMID- 9411148 TI - [Two cases of Creutzfeldt-Jakob disease. Complex symptomatology makes the diagnosis more difficult]. PMID- 9411150 TI - [A drunk person had right to refuse care]. PMID- 9411149 TI - [Growth factors in the uterus are surveyed]. AB - The central localisation of the uterus in the female body may be seen as reflecting its central function in human reproduction, though in rare cases pregnancy may be established even in the absence of uterus. The sex steroids are largely responsible for the central regulation of uterine function, and recent productive research has shown the significance of peptide growth factors for various physiological functions, including the development and repair of the endometrium during the menstrual cycle, as well as the adaptation and growth of the organ during pregnancy. The possible involvement of angiogenic peptides in endometrial neovascularisation is discussed, as is the significance of proto oncogenes and growth factors for the development of uterine fibroids. Further development in this field may have a variety of clinical implications, including the possibility of promoting or inhibiting implantation by manipulation of endometrial angiogenesis. PMID- 9411151 TI - [Growing waiting lists for transplantation in Sweden. Experiences from abroad improved organ retrieval]. PMID- 9411152 TI - [Price of the welfare and physicians' role in the debate on sterilization. Strong reactions in the USA to our treatment of "the weak ones in the society"]. PMID- 9411155 TI - [The Lydia home is an oasis for cancer patients. Here the needed support is offered]. PMID- 9411153 TI - [Compensate victims of condemned therapeutic methods!]. PMID- 9411154 TI - [Per Mindus on an intricate question in a current debate: involuntary sterilization and lobotomy--who should receive monetary compensation?]. PMID- 9411156 TI - [Is the role of authorities never difficult?]. PMID- 9411158 TI - [Diagnostic defaitism worsens the pressure on patients with dyslexia!]. PMID- 9411157 TI - [Is the concept of disease limitless?]. PMID- 9411159 TI - [Instant thrombolysis in stroke]. PMID- 9411160 TI - [Concordance between general psychiatry and forensic psychiatry--objectivity is necessary]. PMID- 9411161 TI - [Medical learning technology--ambitions and results]. PMID- 9411162 TI - [Genetic screening discovers hemochromatosis. Organ damage caused by iron may be prevented]. PMID- 9411163 TI - [Hereditary hemochromatosis. Genetic findings result in new therapeutic possibilities]. AB - Hereditary haemochromatosis is a well-known disease characterised by abnormally high iron absorption and a variety of clinical manifestations. As it's expression is dependent on such factors as gender and diet, it has often been difficult to diagnose before the onset of damage to different organs. However, since the recent discovery of the gene for hereditary hemochromatosis, early detection and treatment of the disease has become possible. PMID- 9411164 TI - [Order, discipline and orderliness were supposed to help mental patients]. PMID- 9411165 TI - [Decreasing number of orchiectomies in prostatic cancer. Results of 9-year registration of the disease are reviewed]. AB - There are several controversies regarding the management of prostate cancer. Whether curative treatment (e.g. radiotherapy and total prostatectomy) prolongs survival remains uncertain. Conservative management is beset with such unresolved issues as the effect on tumour progression of early instituted hormonal treatment and of treatment deferral. Since the outcome of the different types of treatment is unclear the value of screening and early detection remains uncertain. Owing to such issues as these, and the divergent views on prostate cancer, there is an urgent need of auditing. To co-ordinate prostate cancer care in the South-east Region of Sweden, a local management programme with principles for investigation and treatment has been used since 1987. An important feature of the programme is a register containing information on all cases of prostate cancer in the region, including the patient's national registration number, date of diagnosis, basis of diagnosis (cytology, pathology), tumour stage, histological grade, first line treatment, and date and cause of death. Secondary treatment has also been recorded since 1990, and treatment with prostate specific antigen since 1992. From 1987 to 1995, a total of 5,939 cases of prostate cancer were registered, the annual total increasing from 531 to 779 in 1993, after which there was a slight overall decrease but a small increase in the proportion of local tumours. The proportion of incidental tumours followed the same pattern as the transurethral prostatectomy rate in the region. M- and N-categorisation were done to a greater extent in the under-70 than in the over-70 age group. Orchidectomy is rapidly being replaced by treatment with GnRH (gonadotrophin-releasing hormone) analogues. Over the 9-year period, the total prostatectomy rate decreased from 12.5 to 4.6 per cent. All units responsible for prostate cancer care in the south east region regularly receive processed updates from the register providing information on diagnostic and therapeutic methods used and their effect on mortality, thus providing a basis for improving the quality of prostate cancer care. Starting in 1997, a similar registration system is to be extended to cover the entire country. PMID- 9411166 TI - [A vacation substitute in Tanzania. 200 beds for 477 malaria patients]. PMID- 9411167 TI - Viral illness and chronic fatigue (syndrome) PMID- 9411168 TI - [Morphologic parameters for quantitative determination of inflammatory activity of the peritoneum]. AB - The morphology of the inflammatory activity of the peritoneum has been measured qualitatively but quantitative assessments are not common. In a standardized rat model we induced chronic abscess-forming peritonitis after laparotomy and inoculation of 2 ml Bacteroides fragilis suspension at a concentration of 10(9)/ml colony-forming units. The morphological inflammatory activity was determined quantitatively by staining the specimen of the peritoneum with naphthol-AS-D-chloracetate-esterase (NASDCE); through this staining the cytoplasm of granulocytes and tissue mast cells were marked. The peritonitis group (n = 53) and controls (n = 15) were randomly divided into three subgroups (nPeritonitis = 17/18/18 vs. ncontrol = 5/5/5) and observed for 3/7/14 days, respectively. On days 3/7/14 we diagnosed intra-abdominal abscesses in 2 of 17, 13 of 18, and 12 of 18 animals in the peritonitis group. In controls there were no abscesses (P < 0.05). The total cellularity and NASDCE-positive rates on days 3/7/14 in the peritonitis group were 301/409/280 (vs. 155/240/273 in controls) and 1.8/2.9/3.6% (vs. 0.7/0.9/1.4%) in the non-abscess-forming regions and 392/661/625 and 14.4/12.9/11.5% in the abscess-surrounding regions in the infected animals, respectively (P < 0.05). We conclude that the qualitative histological evidence of the morphological inflammatory activity of the peritoneum in the form of an abscess can be supplemented by a quantitative method. Through NASDCE staining the granulocyte and tissue mast cell proportion of the total cellularity as main indicators of the local inflammatory activity can be estimated in peritonitis. This method can be helpful in deciding when to definitively close the abdomen in the course of a programmed lavage treatment in peritonitis. PMID- 9411169 TI - [Surgical management of dysfunctions of dialysis fistulas]. AB - Due to the superficial position of shunt vessels we do not use complicated equipment or diagnostic procedures in the morphological assessment of shunt insufficiency or shunt occlusion. Preoperatively, we merely conduct a clinical examination including inspection, pulse, palpation of the shunt veins and arteries with and without venous congestion, and shunt auscultation. Subsequently, we reoperate the shunt under local anesthesia, at which time the anastomosis is usually checked and repositioned. From January 1995 to May 1996, 539 shunt operations were performed in 371 patients, whereby 263 of these were reoperations. The reoperations were performed due to shunt occlusion (n = 144), shunt stenoses (n = 60), shunt aneurysms (n = 17), steal syndrome (n = 3), and rare complications such as hematoma, shunt infection, seroma, and other disturbances (n = 6) (32 patients were treated in other clinics after reoperation or the functional disturbance of the shunt was not recorded). Angiography was only conducted if the clinical examination did not provide enough information about the shunt problems, and so, preoperatively, only six angiographic examinations were conducted (stenosis, n = 3; aneurysm, n = 1; steal syndrome, n = 2). All reoperations, with only few exceptions (PTFE shunt), were conducted under local anesthesia. At reoperation, 184 new proximal shunts were made, 14 thrombectomies conducted, seven PTFE fistulas made, 13 shunts positioned on the opposite side, five shunts ligated, and eight various other operations performed (32 patients were given further treatment elsewhere or no treatment records were available). If during reoperation flow disturbances were suspected (arterial stenosis) or the blood was flowing towards center (proximal venous stenosis) angiography was performed intraoperatively to assess the condition of the vessels. The 4% rate of early occlusion using this procedure was very low. Only 21 patients had to have more than two reoperations. After 2 years 65% of the reoperated AV fistulas were still functional. Without further diagnostic procedures, we performed immediate, outpatient reoperation under local anesthesia, preferably positioning new proximal shunts so that dialysis could be conducted immediately using the existing dialysis shunt. Only if there were particularly complex functional shunt disturbances (steal syndrome, proximal venous flow disturbance, or arterial stenosis) did we employ other diagnostic procedures (angiography, DSA). With this approach the functional shunt disturbances could be eliminated quickly and effectively, which also minimized the cost and stress for the patient. PMID- 9411170 TI - [Systemic release of prostanoids after surgically-induced injury of lung tissue]. AB - In a prospective study, the systemic inflammatory consequences of surgery-induced lung tissue injury were evaluated using biochemical markers. The aim was to examine whether this type of injury produces a specific pattern of prostanoid plasma levels (prostacyclin, thromboxane, PGE2, PGF2 alpha, and PGM). We, therefore, compared 18 patients (group 1) who underwent thoracotomy without injury to the lung with 26 patients (group 2) that had a resection of pulmonary tissue due to benign diseases. Group 2 patients clearly revealed increased plasma levels of C-reactive protein as well as of the granulocyte-specific PMN-elastase. In particular, there was a pronounced release of prostacyclin and its antagonist thromboxane A2 following lung tissue resection. In contrast to group 1 patients, lung tissue damage resulted in immediately elevated plasma levels of PGF2 alpha and PGE2. When, however, taking into account the time course of PGM, the stable cleavage product of PGF2 alpha, there was no hint of an altered pulmonary metabolic capacity. Presumably, this pattern of elevated prostanoid levels in group 2 is the result of the surgical damage to the lung tissue. Therefore, it can be suggested to be specific for that type of injury. Thus, the release of prostanoids following surgery-induced lung tissue damage may indicate the importance of these mediators, particularly in thoracic injuries associated with lung damage since those may lead to post-traumatic pulmonary dysfunction. These substances may also be useful in evaluating both the severity and the extent of lung tissue damage following major trauma. PMID- 9411171 TI - [Incidence of contralateral inguinal hernias in infancy and childhood]. AB - Inguinal hernia is a frequent surgical disease during infancy, occurring in 1 to 2% of all mature newborns and rise up to 30% of all premature babies. In 9.5% a contralateral hernia is found after unilateral operation. In our own patients this rate was 5.6%. The development of a contralateral hernia was significantly more often found in boys than in girls. If the hernia occurred during the first two months of life, a contralateral hernia developed later highly significant (p > 0.0001). Within the first two postoperative years the second hernia arose in 84.9%. We recommended to routinely operation for a contralateral hernia in all children younger than two months. PMID- 9411172 TI - [A break or bend in the unreamed tibial intramedullary nail. Experimental study]. AB - The intramedullary tibial nail with a proximal angle according to Herzog was developed in order to facilitate implantation. However, the modified technique of unreamed nailing also required a shift of the point of insertion; as a consequence the proximal angle required a considerable increase in the force necessary to introduce the nail. In a study using-four cadaver bones and five commercially available unreamed femoral and tibial nails, the authors demonstrate this considerable increase in insertion force and the development of pressure in the medullary cavity. The measurements made with our experimental setup clearly show that the proximal angle of the unreamed tibial nails available for our series does not have a favourable influence on insertion behaviour. As it appears, it results in an increase in the force required for insertion of the nail, thus also causing a greater strain on the bone and an increase in pressure in the medullary cavity. In contrast, the continuous bend of the nail results in a much smoother course of pressure development in the medullary cavity, which does not reach the same high values as with the unreamed tibial nails, despite the fact that less time is required for insertion of the nail. In our opinion, modification of the axial shape of the nail would result both in better implantability and easier removal. We therefore advocate such a modification of the axial shape of intramedullary tibia nails. PMID- 9411173 TI - [Progress in laparoscopic sigmoid resection in elective surgical therapy of sigmoid diverticulitis]. AB - The full significance of laparoscopic technique in elective surgery of sigmoid diverticulitis has yet to be determined. However, it seems worthwhile to evaluate how minimally invasive surgery could be integrated into the surgical treatment of diverticulitis disease. Between January 1995 and August 1996, 26 patients with sigmoid diverticulitis underwent elective surgery. Following diagnostic laparoscopy, seven patients were treated with primary conventional resection, 15 patients with laparoscopic resection and four patients with laparoscopic-assisted surgery. One laparoscopic resection had to be converted to a median laparotomy. Postoperative complications (n = 2) only appeared in the group of conventional resections. Conventional resections required less time than laparoscopic or laparoscopic-assisted resections, but postoperatively, patients with laparoscopic resection were able to defecate sooner and required a shorter hospital stay. For 60% of the patients with diverticulitis disease of the colon, elective laparoscopic resection may prove to be the best alternative of surgical treatment. In selected patients it is a sound technique with a low complication rate. We recommend that all patients with diverticulitis disease requiring elective surgery undergo diagnostic laparoscopy to determine whether or not laparoscopic resection is a viable option. PMID- 9411174 TI - [Benign symmetrical lipomatosis. A therapeutic dilemma?]. AB - Benign symmetric lipomatosis (BSL), also known as Madelung's disease, is a rare condition and characterized by diffuse but painless growth of unencapsulated lipomas. A close correlation to alcohol and nicotine abuse, metabolic disturbances and malignant tumours have been observed. Surgical treatment is frequently followed by recurrence, nevertheless, it can yield satisfactory functional and cosmetic results. A case of BSL with uncommonly distributed tumors is reported. PMID- 9411175 TI - [Cholecystectomy by mini-laparotomy with the Jako retractor system]. AB - In recent years, minimally invasive operations in abdominal and thoracic surgery, especially with laparoscopic techniques, have developed rapidly. A multi-purpose retractor system, Jakoscope (Atlantis Surgical, Inc., New Brunswick, NJ, USA), developed by Jako provides a new and interesting alternative for minimally invasive and direct access surgery without pneumoperitoneum. This instrumentarium, apart from its use in various surgical specialties, allows a direct access through a 3- to 6-cm single incision for removal of the gallbladder. We used this instrumentarium with great success in patients not suited for laparoscopic surgery in comparable time. Up to now the Jakoscope has been applied in five patients. We report about our first case. PMID- 9411176 TI - [Disappointments and trends]. PMID- 9411177 TI - [Progress in obesity research]. PMID- 9411178 TI - [Proteus infections]. PMID- 9411179 TI - [Prevention of dental caries and periodontitis. Healthy teeth for a lifetime (2)]. PMID- 9411180 TI - [The guaranteeing of quality and safety in fish production--the basis for the prevention of biohelminthiases in the population of Russia at the current stage]. PMID- 9411181 TI - [The spread of Opisthorchis invasion in Voronezh Province]. PMID- 9411182 TI - [Enhancement of the efficacy of Bacillus sphaericus during its encapsulation by Tetrahymena pyriformis infusorians]. PMID- 9411183 TI - [An assessment of the state of scabies morbidity in Russia and the means for its treatment]. PMID- 9411184 TI - [Current problems in disinfection technics in the prevention of infectious diseases]. PMID- 9411185 TI - [The role of the infectious factor in pathology of the gastrointestinal tract]. PMID- 9411186 TI - [Current problems in the under- and postgraduate training of epidemiologists and parasitologists]. PMID- 9411187 TI - [Experience with the practical use of a comprehensive estimation of the level of infectious and parasitic morbidity during the implementation of epidemiological surveillance by the State Epidemiological Health Surveillance Center in the Taganrog area]. PMID- 9411188 TI - [An assessment of the state of pediculosis morbidity in Russia and the modern means for its control]. PMID- 9411189 TI - [The characteristics of the circadian rhythm in the activities of Ixodes ricinus ticks on the Kurskaia Kosa (Kaliningrad Province)]. PMID- 9411191 TI - [Human trichinelliasis in Transcarpathia (1946-1996)]. AB - The paper presents the results of analysis of the epizootic and epidemiological situation associated with Trichinella infection in Transcarpathia. Nine trichinosis outbreaks in 1984-1996, afflicting 132 individuals, were examined. There was an increase in the number of outbreaks and infected men. The bulk of infection is associated with the intake of wild animal meat. Wild boar meat heads the list of sources of infection. PMID- 9411190 TI - [An outbreak of trichinelliasis in Vologda Province]. PMID- 9411192 TI - [Experience in the eradication of foci of geohelminthiases in Sumy Province]. PMID- 9411193 TI - [The prevalence of parasitic diseases and their prevention in Russia]. PMID- 9411194 TI - [The status of the problem of pediculosis in the world]. PMID- 9411195 TI - [Parasitic zoonoses transmitted via environmental objects and food products (meat, fish, etc.)]. PMID- 9411196 TI - [The search for agents with a nontraditional mode of action for the disinfection of environmental objects in parasitoses]. AB - The ovicidal effects of 3 agents: hydrogen peroxide, pantocidum, and Bingsti derived from plant materials (Russia's patent No. 2062752 in 1996) were tested. The findings suggest that pantocidum and hydrogen peroxide were ineffective when used in the tested doses for decontamination. Bingsti was promising in disinfecting sewage. The usage of the nontoxic plant-based agent to decontaminate environmental objects will solve the problem in the utilization of sewage deposits and reduce the risk of parasitosis in human beings. PMID- 9411197 TI - [Still's syndrome in the adult. A report of 8 cases with special reference to diagnostic value of ferritin]. AB - BACKGROUND: Adult onset Still's disease (AOSD) is an uncommon, systemic, inflammatory disorder of unknown etiology, characterized by the triad of fever, arthritis and rash. PATIENTS AND RESULTS: We describe 8 cases of AOSD (3 male, 5 female) diagnosed and treated in the Department of Rheumatology from 1980 to 1996. The delay in reaching a firm diagnosis was between 2 and 86 months, due to both lack of specific serum markers and the abundance of possible differential diagnoses. Our therapeutic strategies and results are presented and the value of obtaining serum ferritin levels for both diagnosis and follow-up studies is discussed. The patients data are compared to those of the world's literature on AOSD. CONCLUSION: The differential diagnosis of fever of unknown origin should always include AOSD, because these patients could be spared from invasive and unnecessary diagnostic measures. Increased serum ferritin levels are of particular value in the diagnosis of acute AOSD and the normalization of the serum ferritin value is a reliable indicator of therapeutic success. PMID- 9411198 TI - [Incidence, clinical picture and treatment of hypothyroid coma. Results of a survey]. AB - BACKGROUND: Myxedema coma is a severe life-threatening clinical state with a high mortality rate. Very often symptoms are masked because of concurrent illnesses. There are no data available about the incidence and prevalence of this disease. Therefore we conducted a survey in the Federal Republic of Germany between 1993 and 1995 by a questionnaire on the occurrence of myxedema coma. METHODS: Questionnaires were mailed to 800 departments of medicine. RESULTS: We received 168 questionnaires for further evaluation. Among those, 24 patients were classified as myxedema coma, but according to clinical data we could reclassify 12 patients as myxedema coma and 12 patients as severely hypothyroid but without coma. The mean age of the patients was 73 years. The etiology was Hashimoto's thyroiditis in 16 patients (67%), in 15 patients the thyroid disease was unknown. In 6 patients thyroid hormone therapy was withdrawn after thyroid surgery. One patient became hypothyroid after radioiodine therapy and 1 patient had secondary hypothyroidism. 19 of the 24 patients received i.v. thyroxine therapy and 11 patients received corticosteroids additionally. Six patients (25%) died. CONCLUSION: These data emphasize that myxedema coma is a rare disease (24 patients within two years in Germany) occurring especially in older patients and is associated with a high mortality rate also in non-comatose patients. In the majority of the patients myxedema coma was the first manifestation of thyroid disease. PMID- 9411199 TI - [Diagnosis of pancreatic carcinoma]. PMID- 9411200 TI - [Painless jaundice]. PMID- 9411202 TI - [Transplantation of hematopoietic stem cells. II: Indications for transplantation of hematopoietic stem cells after myeloablative therapy]. AB - The destruction of hematopoiesis and lymphopoiesis by total body irradiation or high dose chemotherapy for the treatment of malignancy can be reversed by the transplantation of allogeneic or autologous hematopoietic stem cells. In primary disorders of bone marrow or immune system, allogeneic stem cells replace deficient cells. Acute leukemias can be cured, with in 50 to 80% disease free survival after 5 to 8 years. The allogeneic graft versus leukemia effect by immunoreactive cells reduces the relapse rate in myeloid and lymphoid malignancies. 40 to 70% of patients with chronic myeloid leukemia remain disease free after more than 5 years. Patients with malignant lymphoma have a 40 to 70% chance of cure with autologous transplantation, which is not increased by allogeneic cells, because of a higher incidence of severe complications. An increasing number of patients without option for cure is treated with the aim of prolonging remission or retarding disease progression, such as in chronic myeloid leukemia, multiple myeloma and certain solid tumors. New studies suggest in breast cancer with axillary lymph node metastases, that adjuvant high dose chemotherapy with autologous stem cell support will significantly improve disease free survival from 30 to over 60% after 3 to 5 years. In congenital metabolic and storage diseases deficient enzymes are substituted by the allogeneic cells. Clinical trials explore the use of stem cell transplantation after myeloablative therapy in autoimmune disorders as well as in gene therapy with transfected hematopoietic stem cells. PMID- 9411201 TI - [Endocrinologic and metabolic complications of Alagille syndrome]. AB - BACKGROUND: Patients with gastrointestinal and hepatobiliary disorders, either congenital or acquired early in childhood, are at high risk for various endocrine and metabolic abnormalities. CASE REPORT: A 27-year-old woman with Alagille's syndrome presented with progressive jaundice and gait disturbances following surgery and ingestion of oral contraceptives. On physical examination, short stature, facial dysmorphism and neuromuscular symptoms such as polyneuropathy and spinocerebellar ataxia were noted. Serum concentrations of total bilirubin (54 mg/dl) and alkaline phosphatase were markedly increased, whereas serum levels of haptoglobin, zinc, vitamin D and E were decreased. Although prehepatic or intrahepatic etiologies of jaundice were more likely in this patient, posthepatic etiologies were ruled out by abdominal ultrasound and endoscopic retrograde cholangio-pancreaticography. Based on a working diagnosis of acute drug-induced cholestasis, treatment with high doses of lipid-soluble vitamins and ursodeoxycholic acid was initiated. In response to therapy, her abnormal laboratory results normalized and her neurologic symptoms markedly improved. CONCLUSION: This clinicopathological conference of a patient with Alagille's syndrome illustrates the clinical presentation and therapy of metabolic and endocrine complications in chronic cholestasis. PMID- 9411203 TI - [Status of digitalis in therapy of acute and chronic heart failure]. AB - Although supported by more than 200 years of experience and anecdotal clinical evidence, the efficacy of digitalis in the management of heart failure has been questioned until the past decade. The idea to improve contractility of the diseased myocardium with an inotropic agent is fundamental in the management of left ventricular dysfunction. The majority of clinical trials published since 1980, most of which examined patients with mild to moderate heart failure, indicate that digitalis alone or in combination with vasodilators may improve the clinical outcome particular in those patients with more advanced symptoms and poorer left ventricular function. Aside from its action as an inotropic drug the pharmacology and the mechanisms by which digitalis influence the diseased myocardium and peripheral circulation in heart failure has gained more complexity within the last years, raising the idea of other mechanisms that might be involved in its action. Particular for ACE inhibition multiple clinical trials have conclusively demonstrated its impact on survival and morbidity in congestive heart failure. Improvement of clinical outcome as measured in terms of fewer hospitalizations and improvement of symptoms in patients receiving digitalis seems to be comparable to patients receiving beta-blockers additional to diuretics and ACE inhibitors, an entirely different approach to the treatment of heart failure. Despite initial improvement of hemodynamics it now appears that there is no survival benefit found for digitalis in the management of heart failure. PMID- 9411204 TI - ["Medical Ethics Patient Forum". Results of a model project]. AB - BACKGROUND: The patients' forum Medical Ethics is an innovative model project, which aims at intensifying the dialogue between health care professionals and patients as well as their relatives. METHODS AND RESULTS: Practical experiences and data from a complementing evaluative study are reported. By means of a sophisticated instrument of moral psychology, the Moralisches-Urteil-Test (M-U T), differences are revealed between the participating groups--doctors, health care professionals, patients, medical students etc.--referring to more cognitive or emotional orientations in moral judgment, respectively. These differences contradict common assumptions. CONCLUSION: On the ground of the data presented as well as by subjective estimations of participants it is shown that the ethics discussion with patients is not only successful, but that it is also valuable for teaching by fostering new insights and experiences. PMID- 9411205 TI - [The knockout mouse--a revolutionizing model in biomedical research]. PMID- 9411206 TI - [Brucellosis with fatal endotoxic shock]. AB - BACKGROUND: Brucellosis is a zoonosis with good prognosis in cases of early diagnosis. To make the diagnosis is still a problem today. CASE REPORT: A 60-year old butcher was admitted with undulating fever, sweats, arthralgia and weight loss. Further examination revealed hepatosplenomegaly with laboratory findings of a hepatitis and multiple focal liver lesions shown by abdominal ultrasound and CT. Histologically, these lesions corresponded to caseous granulomas. Diagnosis of brucellosis was confirmed by detection of brucella species in prolonged incubation in blood culture. After the beginning of antibiotic resistance-tested therapy with tetracycline and quinolones, an endotoxic shock occurred during the first 24 hours of treatment and the patient died after multiorgan failure with disseminated intravascular coagulation. CONCLUSION: In cases of undulating fever with liver involvement, a brucellosis should be considered. Good teamwork of the internal, pathological and microbiological departments is necessary for early and correct diagnosis. This is the first report of human brucellosis in association with lethal endotoxic shock. PMID- 9411207 TI - [Differences between evidence based-medicine and best conventional medicine]. AB - The practical application of rational concepts will be possible in the health care system if two conditions will be met. The legal framework has to fit these goals and new strategies are to establish. The new strategies will be needed to identify from the huge amount of data "evidence" which will support the medical progress. Evidence-Based Medicine (EBM) offers these strategies. Modifications of our conventional procedures are necessary to establish EBM: the conditions of clinical actions, the definitions of goals of medical training, the rational of clinical decisions and the continued medical education. PMID- 9411208 TI - [Diagnostic importance of middle latency auditory evoked potentials (MLAEP) in multiple sclerosis]. AB - This study was aimed at assessing the interest of middle latency auditory evoked potentials (MLAEP) recordings in multiple sclerosis (MS). Brainstem auditory evoked potentials (BAEP) and MLAEP were recorded in 40 control subjects and 65 patients who had MS according to Poser's criteria. Na and Pa latencies were significantly delayed in the MS group. These abnormalities were detected in 60% of the patients. In 26% of the cases with normal BAEP, abnormalities of MLAEP were present. In 71% of the patients abnormalities of both BAEP and MLAEP were observed. These results suggest the ability of MLAEP recordings to detect lesions of the auditory pathways rostral to the pons and show the value of their combined recordings in MS patients. PMID- 9411209 TI - [Recent changes in therapy of pancreatic cancer]. PMID- 9411210 TI - [Relationship between postoperative liver dysfunction following stomach cancer operation and gene deletion of glutathione S transferase]. PMID- 9411211 TI - [Preclinical study on method of recombinant adenovirus gene transfer to liver cells]. PMID- 9411212 TI - New paradigms and new issues: a comment on emerging themes in the study of motor development. PMID- 9411213 TI - [Receptor-antireceptor interaction in viral infections]. AB - Discusses the problem of virus-cell interactions. Characterizes the virus attachment proteins (antireceptors) and virus receptors on the cell membrane. Analyzes some mechanisms of the receptor-antireceptor reactions during a virus infection. PMID- 9411214 TI - [Development of polymerase chain reaction-based test systems for detecting leptospira in polytypical leptospirosis foci]. AB - Two highly sensitive test systems G and B, based on the polymerase chain reaction, were developed for indication of pathogenic Leptospira interrogans, including the serovariants appearing during outbreaks in polytypical foci of leptospirosis in the tropical zone of China. These test systems can be used for rapid diagnosis of leptospirosis in humans in foci with different etiological structure. PMID- 9411215 TI - [Preparation of new microsatellite markers of rat chromosome 10 in the region, containing genes regulating blood pressure level]. AB - Primer walking is a simple and efficient technique to find new microsatellites in the regions adjacent to known sequences. The method was applied to obtaining new microsatellite markers for rat chromosome 10 in the region of a blood pressure quantitative trait locus. Eight microsatellites were found, five of them were polymorphic in at least one population used for mapping. Improved genetic maps of this region for F2(SxMNS) and F2(SxLEW) populatons were constructed. PMID- 9411216 TI - [Targeting the gene for platelet-derived growth factor A-chain (PDGF-A) and preparation of chimeric mice with a "knockout" genome]. AB - A vector was selected for homologous recombination to generate mouse embryonic stem cell clones with disrupted platelet-derived growth factor A (PDGF-A) chain gene. A chimeric mouse with targeted PDGF-A gene has been created. PMID- 9411217 TI - [Use of a phage peptide library in mapping the group-specific hemagglutinating domain of alpha-virus glycoprotein E2]. AB - Phage display peptide library f88-4/15 (G. P. Smith, USA) was used for mapping the hemagglutination activity domain of glycoprotein E2 of alphaviruses. Using affinity selection and ELISA, we selected the clones binding monoclonal antibody 4H5 to Venezuelan equine encephalomyelitis virus and inhibiting alphavirus hemagglutinating activity. Analysis of the similarity between the peptides amino acid sequences with the alphavirus glycoprotein E2 sequences revealed a structural motive of 4 amino acid residues (HTSR) which was identified in the 85 88 region. Bacteriophages F36 and F19 contained motives corresponding to 102-SXXM 105 and 109-AXXP-112 regions in alphavirus proteins E2. These data permit us to propose that the detected regions are fragments of a group-specific alphavirus hemagglutination domain. PMID- 9411218 TI - [Isolation and analysis of two cDNA-clones expressing the human lymphocyte expression library, inducing DNA-binding activity in vitro]. AB - Two cDNA clones are sequenced which were isolated from human lymphocyte expression library using Southwestern (DNA-binding) screening with 32P-labeled Alu DNA in the presence of 100-fold excess of unbalanced poly (dI-dC). In one of the sequenced clones (vb22) an open reading frame (ORF) is detected, encoding protein with a new potential DNA-binding (zink-finger) domain, and DNA-binding activity of the protein is directly confirmed after its expression (as GST-fusion protein) in Escherichia coli. The other sequenced clone (wa12) is partially homologous to 15EST sequences present in GENBANK (April, 1996) and cloned from very different human tissues. Connection of these 15 overlapping GENBANK sequences resulted in a longer sequence covering wa12 and having ORF potentially encoding a new 10 kDa polypeptide without any apparent DNA-binding domains. This connected sequence as well as wa12 sequence having only 65 amino acids ORF are unrecognizable by computer software as the protein-coding regions, and we suppose that wa12 transcripts possess DNA-binding activity. Homopyrimidine blocks in RNA longer than 12 nucleotides are known to bind mirror duplex DNA sequences to form triplexes whose stability is comparable to that of protein-DNA complexes, and human promoters contain many such blocks. PMID- 9411219 TI - [Nonhistone chromosome proteins HMG1 and HMG2 stabilize one of the sequence specific complexes, formed on the promotor of human retroposons of the ALU-family of other nuclear proteins]. AB - Human retroposons of the Alu family have an internal promoter for RNA polymerase III also present in tRNA genes, but Alu are poorly transcribed by this polymerase in somatic cells in vivo, which is probably due to an efficient system of repression of the Alu transcription. The key control promoter element of the tRNA genes, known as B-box, binds basal transcription factor TFIIIC2, which initiates assembly of the full transcription complex. Previously we identified several human nuclear factors which bind to Alu Subregion covering B-box and the adjacent sequences, which can be involved in repression of Alu transcription. In this study we identified one factor, F1, as HMF1/2 and another factor, F2, as TFIIIC2 like protein reacting with B-box. Both the factors are not alu-specific, whereas the third identified Alu-binding factor, F3, seems to be specific for the Alu specific subsequence located just downstream from B-box and can discriminate evolutionary "young" and "old" Alu subfamilies producing different numbers of transcripts in vivo. Although HMG1/2 binds to Alu in a sequence-unspecific manner, the proteins are capable of stabilizing the sequence-specific complex F3 which can be functionally significant. We believe that the factors binding to Alu B-box subregion interfere somehow with the functions of the basal factor TFIIIC2 and, hence, can be co-repressors of Alu transcription by RNA polymerase III. PMID- 9411220 TI - [DNA polymorphism in the region of APOB100, APOCIII, APOE, and angiotensin converting enzyme genes and indicators of the lipid spectrum in children and adolescents in St. Petersburg]. AB - Polymorphisms of 3 apolipoprotein genes Xba I apoB, Sstl apoCIII, and apoE and the insertion-deletion polymorphism of the angiotensin-converting enzyme gene (I/D ACE) and lipid levels were studied in a random sample of 403 children and adolescents aged 6 to 18 years living in St. Petersburg. The children were divided in 4 age groups with consideration for the relative body weight index: group 1.6 to 9 years; II, 10-12; III, 13-15; and IV, 16-18 years. The first three groups were divided by sex, the fourth was not because it was the smallest. Relationships between lipid levels and DNA polymorphisms of the above genes were analyzed in all groups. Effects of apoB Xbal, apoCIII Sstl, apoE, and ACE genotypes on the levels of the blood basic lipids were analyzed using Statgraphics software. A marked effect of the apoE (E3/E4) genotype on the total and LDL-cholesterol variability was observed in group IV. The individuals carrying the E4 apoE allele had increased levels of total and LDL-cholesterol (p < 0.02 and p < 0.03, respectively). The level of triglycerides was higher in the subjects carrying the S2 apoCIII allele in the third group (p < 0.04). A statistically reliable difference was however observed only in girls (p < 0.01). We failed to detect reliable correlations between lipid levels and various apoB and ACE genotypes. Hence, the genetic variants of apoCIII and apoE genes affect the blood lipid levels as early as in adolescence. PMID- 9411221 TI - A comparison of antiarrhythmic-drug therapy with implantable defibrillators in patients resuscitated from near-fatal ventricular arrhythmias. AB - BACKGROUND: Patients who survive life-threatening ventricular arrhythmias are at risk for recurrent arrhythmias. They can be treated with either an implantable cardioverter-defibrillator or antiarrhythmic drugs, but the relative efficacy of these two treatment strategies is unknown. METHODS: To address this issue, we conducted a randomized comparison of these two treatment strategies in patients who had been resuscitated from near-fatal ventricular fibrillation or who had undergone cardioversion from sustained ventricular tachycardia. Patients with ventricular tachycardia also had either syncope or other serious cardiac symptoms, along with a left ventricular ejection fraction of 0.40 or less. One group of patients was treated with implantation of a cardioverter-defibrillator; the other received class III antiarrhythmic drugs, primarily amiodarone at empirically determined doses. Fifty-six clinical centers screened all patients who presented with ventricular tachycardia or ventricular fibrillation during a period of nearly four years. Of 1016 patients (45 percent of whom had ventricular fibrillation, and 55 percent ventricular tachycardia), 507 were randomly assigned to treatment with implantable cardioverter-defibrillators and 509 to antiarrhythmic-drug therapy. The primary end point was overall mortality. RESULTS: Follow-up was complete for 1013 patients (99.7 percent). Overall survival was greater with the implantable defibrillator, with unadjusted estimates of 89.3 percent, as compared with 82.3 percent in the antiarrhythmic drug group at one year, 81.6 percent versus 74.7 percent at two years, and 75.4 percent versus 64.1 percent at three years (P<0.02). The corresponding reductions in mortality (with 95 percent confidence limits) with the implantable defibrillator were 39+/-20 percent, 27+/-21 percent, and 31+/-21 percent CONCLUSIONS: Among survivors of ventricular fibrillation or sustained ventricular tachycardia causing severe symptoms, the implantable cardioverter-defibrillator is superior to antiarrhythmic drugs for increasing overall survival. PMID- 9411222 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 37-1997. A 59-year-old man with anorexia, weight loss, and a mediastinal mass. PMID- 9411223 TI - Prednisone and aspirin in women with recurrent fetal loss. PMID- 9411224 TI - Antiphospholipid antibodies, annexin V, and pregnancy loss. PMID- 9411225 TI - Antiphospholipid antibodies, annexin V, and pregnancy loss. PMID- 9411226 TI - Plasma homocysteine levels and mortality in patients with coronary artery disease. PMID- 9411227 TI - Plasma homocysteine levels and mortality in patients with coronary artery disease. PMID- 9411228 TI - Plasma homocysteine levels and mortality in patients with coronary artery disease. PMID- 9411229 TI - Fungal sinusitis. PMID- 9411230 TI - Fungal sinusitis. PMID- 9411231 TI - Interferon alfa-2b and cytarabine in chronic myelogenous leukemia. PMID- 9411232 TI - Is psychoanalysis science? PMID- 9411233 TI - False positive dipstick test for malaria. PMID- 9411234 TI - Malaria acquired 13 times in two years in Germany. PMID- 9411235 TI - Treatment of prostate cancer with goserelin and radiotherapy. PMID- 9411236 TI - Treatment of prostate cancer with goserelin and radiotherapy. PMID- 9411237 TI - End-tidal carbon dioxide and outcome of out-of-hospital cardiac arrest. PMID- 9411238 TI - End-tidal carbon dioxide and outcome of out-of-hospital cardiac arrest. PMID- 9411239 TI - Resuscitation medicine. PMID- 9411240 TI - Isoniazid prophylaxis for high-risk patients with anergy and HIV infection. PMID- 9411241 TI - Isoniazid prophylaxis for high-risk patients with anergy and HIV infection. PMID- 9411242 TI - Treatment of early syphilis. PMID- 9411244 TI - Necrotizing fasciitis due to group A streptococcus after an accidental needle stick injury. PMID- 9411243 TI - Treatment of early syphilis. PMID- 9411245 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 38-1997. Inflammation of the ears, anemia, and fever 21 years after treatment for Hodgkin's disease. PMID- 9411246 TI - Valvular heart disease associated with fenfluramine-phentermine. PMID- 9411247 TI - Valvular heart disease associated with fenfluramine-phentermine. PMID- 9411248 TI - Valvular heart disease associated with fenfluramine-phentermine. PMID- 9411249 TI - Valvular heart disease associated with fenfluramine-phentermine. PMID- 9411250 TI - Valvular heart disease associated with fenfluramine-phentermine. PMID- 9411251 TI - Valvular heart disease associated with fenfluramine-phentermine. PMID- 9411252 TI - Valvular heart disease associated with fenfluramine-phentermine. PMID- 9411253 TI - Digital necrosis associated with dexfenfluramine. PMID- 9411254 TI - Hematopoietic stem-cell transplantation for systemic lupus erythematosus. PMID- 9411255 TI - Fatal esophagoaortic fistula after placement of a self-expanding metal stent in a patient with esophageal carcinoma. PMID- 9411256 TI - Correction of endnote in ProPAC report. PMID- 9411257 TI - Costs at for-profit and at not-for-profit hospitals. PMID- 9411258 TI - Costs at for-profit and not-for-profit hospitals. PMID- 9411259 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 39-1997. A 67-year-old woman with the cauda equina syndrome. PMID- 9411260 TI - Correction and revision of conclusions--dietary trends in the United States. PMID- 9411262 TI - Transmission of hepatitis C virus during colonoscopy. PMID- 9411261 TI - Correction and revision of conclusions--dietary trends in the United States. PMID- 9411263 TI - Increased serum prostate-specific antigen in a man and a woman with hepatitis A. PMID- 9411264 TI - Confusion about breast-cancer screening. PMID- 9411265 TI - Confusion about breast-cancer screening. PMID- 9411266 TI - The Medicare-HMO revolving door. PMID- 9411267 TI - Disability and physician-assisted suicide. PMID- 9411268 TI - Disability and physician-assisted suicide. PMID- 9411270 TI - Disability and physician-assisted suicide. PMID- 9411269 TI - Disability and physician-assisted suicide. PMID- 9411271 TI - [Psychosocial risk and protective factors in childhood and adolescence as predisposition for psychiatric disorders in adulthood. Current status of research]. AB - Longitudinal and well-controlled cross-sectional studies have shown that numerous psychosocial stress factors in childhood and youth affect psychic health in the long term. The significance of compensatory protective factors has also been well documented in various studies. An overview of this work is presented: the factors which need to be clarified and taken into consideration in the diagnosis of psychic disorders have been summarized and critically analyzed on the basis of the results of more recent studies of the significance of sexual traumatization. PMID- 9411272 TI - [Systemic immunologic diseases as differential diagnosis in psychiatry]. AB - Psychiatric symptoms may be caused by systemic autoimmune diseases. Quite often, mental disorders are an early symptom during the course of an autoimmune disease and sometimes they may even be the presenting symptom. This article reviews psychiatric and neurologic symptoms in systemic lupus erythematodes, Sjogren syndrome, primary vasculitides and other immunopathies such as the primary antiphospholipid syndrome and Sneddon's syndrome. The article also discusses diagnostic aspects and therapeutic options if an autoimmune disease as cause of a psychiatric or neurologic symptom is suspected. An increased awareness of psychiatrists and neurologists will make it possible that systemic autoimmune diseases are early identified as a possible cause of psychiatric symptoms and may then be treated adequately. PMID- 9411273 TI - [Premenstrual dysphoric disorder. An overview of diagnosis, epidemiology and therapeutic approaches]. AB - Even though premenstrual symptoms had been already described by Hippocrates, premenstrual dysphoric disorder (PMDD) was first mentioned as a special psychiatric diagnosis in the Diagnostic and Statistical Manual of Mental Disorders (DSM-IV) in 1994. In DSM-III-R-Appendix A is was called late luteal phase dysphoric disorder (LLPDD), Appendix A. Before this diagnosis was established based on operationalized criteria, the term premenstrual syndrome (PMS) was used for patients with severe premenstrual mood disturbances and physical symptoms. Many hypotheses about the pathophysiological mechanisms underlying PMS and PMDS led to different therapeutic strategies. While PMS was mainly treated by gynecologists, PMDD became of interest in psychiatric research. Several antidepressants, psychotherapy, sleep deprivation and light therapy have been investigated regarding their effectiveness in combatting premenstrual symptoms such as depression, tension, dysphoria and anxiety. Within the anti depressants the best findings were for selective serotonin reuptake inhibitors (SSRIs). PMID- 9411274 TI - [Dementia in the very elderly. Results of the Berlin Aging Study]. AB - The frequency of dementia in very old subjects, the risk factors and the consequences of the disease were investigated in the Berlin Aging Study in an age and gender-stratified design (ages 70-103 years, n = 516). Psychiatrists diagnosed a dementia syndrome according to DSM-III-R, applying the GMS-A and HAS interviews. The dementia frequency steeply increases until the 90-94 year group, but there is no further exponential increase for the 95+ group--instead for men the data show a plateau of dementia prevalence. Low education level turned out to be a risk factor, which explains the gender effect in a logistic regression analysis. The apolipoprotein E4 genotype was confirmed as a risk factor--however, only for the older subjects (85+). Dementia was a major reason for institutionalization. The 2-year mortality was no higher in dementia than for age matched non-demented controls. The results gave a detailed picture of dementia in the very old. This is a prerequisite for planning facilities for psychiatric diagnostics and therapy as well as nursing care. PMID- 9411275 TI - [Direct sequelae of accidental falls in hospitalized geriatric psychiatry patients]. AB - Falling is an everyday risk for elderly people, with the consequence of getting hurt. What exactly the dangers are, especially in people with gerontopsychiatric disorders, has hardly ever been mentioned in today's literature. Therefore this report deals with 887 documented fall cases in 758 gerontopsychiatric inpatients during a cumulative time period of 3845 months. The consequences of the falls according to sex and diagnosis of dementia were divided in four categories, depending on how serious the injuries were. About 65 to 71% of the patients don't show any direct consequences, about 10% have serious injuries of which 5% are fractures. There are significantly more dement women than dement men who suffer from fractures. Women over eighty years have significantly fewer serious fall injuries than women in their eighties; the same phenomenon can be observed in men concerning light injuries. PMID- 9411276 TI - [German and American consultant psychiatrists evaluate their function. A contribution to quality improvement in consultation psychiatry]. AB - There is an increasing interest in scientific and practical aspects of C/L psychiatry in Germany. The every day C/L work in Germany still differs substantially from the American one. In contrast to Germany and other European countries, C/L psychiatry in the USA has developed into a subspecialty with multiple activities in research and patient care during the last 3 decades. This study was conducted in Germany and the USA in order to evaluate and compare the training and patient care in C/L psychiatry, as assessed by consultants doing the every day C/L work. 227 C/L psychiatrists participated in the study in both countries. There are significant differences in theoretical and practical C/L psychiatry is obviously more comprehensive. American C/L psychiatrists take more time for consultations, do more follow-ups, and more frequently organize psychiatric or psychotherapeutic outpatient after-care. These results are suited to support activities to improve quality in Germany C/L psychiatry. PMID- 9411277 TI - [Assessment of therapy outcome within the scope of quality assurance in a psychiatric clinic. A report of experiences]. AB - A recent modification of German law requires quality assurance of inpatient treatment, i.e., of the quality of therapy, allowing comparison of different hospitals. In this paper a program developed to assess the outcome by means of international, frequently applied psychopathometric scales is presented. This program was evaluated in a study of 258 psychiatric inpatients. This study emphasized the identification of patients with long-term hospitalization and/or worse outcome. A concept to disclose these so-called 'problematic' patients by means of easily administered scales like the SCL90-R, GAF and WHO-Disability Diagnostic-Scale (DDS) has been developed. Patients with a very long hospital stay (> 70 days) show a higher number and more severe symptoms in the self-rating (SCL90-R) at admission than the subjects with a short hospital stay. This pilot study demonstrated that a routine outcome analysis can be used for quality management in psychiatry. PMID- 9411278 TI - [Indications criteria for targeted syphilis screening]. PMID- 9411279 TI - [Metachromatic leukodystrophy simulating schizophrenia-like psychosis]. AB - A case of late onset metachromatic leukodystrophy with a clinical picture of paranoid hallucinatory psychosis and severe dyskinesia is described. The problem of diagnostic recognition is discussed. In the case, diagnostic procedures were initiated after atypical clinical course, and established on the basis of MRI and specific biochemical tests. PMID- 9411281 TI - [Circulatory instability in potential donor organ patients in a state of brain death. Investigation of predictive factors]. AB - OBJECTIVE: To assess the factors influencing and predicting the degree of circulatory impairment of the brain-dead cardiac donors (BDCD). DESIGN: Retrospective study. SETTING: General ICU of a 1400 bed High Specialty Regional Hospital. PATIENTS: 53 patients undergoing brain death (BD) assessment as suitable cardiac-donors. INTERVENTIONS: Fluids were given up to a central venous pressure (CVP) > 5 mmHg and adrenergic drugs in a sequential manner trying to keep the systolic blood pressure > 90 mmHg. METHODS: Every hour during the time of BD assessment were recorded: invasive blood pressure (BP), CVP, fluid balance, the demographic data, the dosage of adrenergic drugs, the cause of admission, the time from admission to ICU to BD, the report of an echocardiogram. The patients were divided into 4 classes according to the dosage of the adrenergic drugs required to achieve the pressure-goal. The 4 classes were compared for the above mentioned parameters. RESULTS: No differences were found among the 4 groups. CONCLUSIONS: In BDCD the degree of circulatory instability and vasopressor needs cannot be anticipated by the parameters examined. This could be due partly to the relative homogeneity of these patients, since they underwent a clinical selection and similar interventions, partly to a possibly inadequate standard of monitoring. Anyway the unique hemodynamic situation after BD is probably not very affected by the previous events. Further studies are necessary to explain the variability of the circulatory instability and to assess its effects on the quality of the transplanted organs. PMID- 9411280 TI - [Coincidence of epilepsy and Asperger syndrome. Case report and review]. AB - Asperger syndrome is an autistic disorder and was first described by Hans Asperger in 1944 without further acceptance in the literature over almost four decades. Following several publications in the 1980s, for the disorder became more widely known and was first introduced into ICD-10 and DSM-IV as a new diagnosis in 1988 and 1994, respectively. The etiology is unknown. We present a female patient with typical features of Asperger syndrome, suffering also from epilepsy and internal medical disorders. The associated diseases, diagnostic criteria and possible therapeutic options are discussed. PMID- 9411284 TI - [Right atrial thrombosis with concomitant thrombus attached to a central venous catheter. Clinical case]. AB - A clinical case of a patient in whom a right atrial thrombosis was casually discovered by transthoracic echocardiography is described. The hypothesis that also the central venous catheter (CVC) could be seat of thrombosis was done, and was confirmed by chest X-ray with dye injection in the CVC. A thrombolytic therapy with plasminogen tissular activator and with heparin infusion was started. After 24 hours from the beginning of therapy instrumental control were performed (transesophageal echocardiography ECOTEE, chest X-ray with dye injection in CVC), showing the completely dissolution of the thrombus. It is observed that, when in a patient with CVC high degree of thrombogenic factors (age over 60 years, presence of cardiac arrhythmias with ventricular hypokinesis, polyglobulia due to chronic obstructive pulmonary disease, CVC, low levels of antithrombin III) are present, the usual prophylactic measures (subcutaneous heparin, hydrophylic catheter) couldn't be sufficient to avoid superior caval vein thrombosis. The conclusion is drawn that these patient should be studied with ECOTEE and eco Doppler. Chest tomography and superior caval venography are also useful its early diagnosis of superior caval thrombosis. PMID- 9411283 TI - [Perioperative thermal homeostasis. A duty of the anesthesiologist]. AB - Anaesthesia, surgical procedures and operating room temperature can deeply alter the human thermoregulatory system. Unexpected and sometimes serious perioperative complications can occur. Many studies have been carried out in order to describe and evaluate the detrimental effects produced by different anaesthesia procedures (whether by general, regional or integrated anaesthesia) on thermic homeostasis. More recently it has also been reported that perioperative hypothermia significantly affects patients' outcome, increasing intraoperative blood losses, incidence of postoperative wound infection, and hospital stay. Italian anaesthetists have still a poor consideration about intraoperative body temperature monitoring and patients' warming as basic important skills for a better anaesthesiologic patients management. According with the literature, we do believe that this is not a right opinion. The purpose of the present paper would be to point out the most important knowledges concerning thermic homeostasis management, in order to increase anaesthesiologist's awareness in this essential field of patients perioperative care. PMID- 9411282 TI - [Severe infections after orthotopic hepatic transplant]. AB - METHODS: The authors analysed severe infections in 43 consecutive patients undergoing orthotopic liver transplant. Prophylaxis and full anti-infection monitoring was performed in all cases. Immunosuppressive therapy was administered in the form of primary cyclosporine in 27 cases and primary OKT3 in 16 cases. RESULTS: Twenty-seven patients are still alive (median 8 months, range 2-40) and 16 died (median 22 days, range 10-92) of whom 4 without and 12 with infection, including two deaths owing to non-correlated causes with infection after recovery. Twenty-three patients underwent 33 episodes of severe infection (plus four with inconclusive positive cultures) without any case of protozoal or viral infection. All episodes occurred within two months of surgery and affected the lung (10), abdomen (7), lung + abdomen (1), urinary tract (1), lung + urinary tract (1), as well as two diffused cases and 7 cases of isolated bacteremia deriving from the donor (1), venous catheters (3), mild otorhinolaryngeal infection (1) and two unknown sources (2). Eighteen infective agents were identified in 45 cases. The bacteria involved in single-agent episodes were: 11 Gram+, 9 Gram- and five fungi. Polymicrobic and bacterial/fungal episodes were repeatedly observed in two and two cases. Postoperative renal insufficiency significantly influenced both the incidence of and mortality due to infection. Overall mortality was also influenced by early graft function, postoperative complications and reoperations, and the incidence of infections by the portal clamping stage, reject and prolonged coma. CONCLUSIONS: The absence of severe viral infections and the gradual reduction of mortality caused by infection appear to be parallel to the aggressive antiviral prophylaxis, the gradual improvement of intra- and postoperative management and primary immunosuppression with OKT3. PMID- 9411285 TI - [Enoximone in the treatment of postoperative low cardiac output syndrome in pediatric heart surgery. Open study in tetralogy of Fallot]. AB - OBJECTIVE: To evaluate the role of the phosphodiesterase inhibitor enoximone in low output states (LOS) following cardiac operations in congenital heart diseases. DESIGN: This was an unblinded, retrospective, open study. Hemodynamic effects of intravenous enoximone were investigated in 130 patients--70 adults and 60 pediatrics--offered to our Department from 1992 to 1995. To avoid multifactorial events due to different cardiopathies, our analysis was limited to 24 newborns and children operated on for correction of tetralogy of Fallot (TOF). SETTING: Cardiac surgery ICU of a Regional Hospital in Italy. METHODS: Retrospective analysis of 24 cases of postoperative LOS in surgical corrected TOF, treated with enoximone, were compared with a control group treated with conventional inotropic drugs. DATA ANALYSIS: Data were compared by "t"-Student's test for impaired data. RESULTS: Significative cardiac function improvement in treated group, demonstrated by an increased systemic pressure values, by decreased right chambers pressures and by significative improvement in oxygen mixed venous saturation. Better hemodynamic recovery after ICU discharge. No significant side effects were detected. CONCLUSIONS: Enoximone is, suggested also in pediatric patients in the management of refractory LOS following open heart surgery, especially when a long-standing treatment is predictable (more than 72 96 hrs). PMID- 9411286 TI - [Use of mechanical staplers in surgery of aneurysms of the abdominal aorta]. AB - This research aims to illustrate the advantages of using staplers in abdominal aorta surgery. The authors describe their experience in a personal case, of a patient suffering from aneurysmatic expansion of the abdominal aorta extending to both the common iliac arteries. The use of staplers for the closing of the distal stumps in particularly advantageous because it makes less problematic the dissection of the iliac and by hypogastric artery from their underlying veins. That involves a risk diminution of the iliac vein accidental lesions, better by hypogastric artery revascularization, shortening of any times and the perfect haemostatic holding of the mechanical suture, allowing a reduction in mortality and morbidity. PMID- 9411287 TI - [Cutaneous mechanical staplers. Our experience]. AB - This paper reports the results of a trial, comparing the use of disposable skin staplers with conventional nylon or silk sutures in skin closure. This study began in 1986; 7274 patients undergoing elective and emergency operations were controlled during the postoperative period, 3 and 6 months after surgery. It was shown that skin clips instead of sutures decrease the operative time, produce wound healing with a good cosmetic results, and above all a significant lowering of the wound infection rate. PMID- 9411288 TI - [Laparoscopic cholecystectomy in patients who previously underwent major laparotomy]. AB - This paper describes the experience gained and the techniques used by the authors in the laparoscopic treatment of biliary calculosis in patients who previously underwent abdominal surgery, excluding cases with incisions below the horizontal line passing through the umbilicus (in these cases there are usually no adhesions to hinder operative laparoscopy of the gallbladder). Between October 1991 and July 1995,776 laparoscopies were performed on bile ducts (715 for calculosis of the gallbladder and 61 for cholecysto-choledochal calculosis). In 18 cases there was scarring of the abdominal wall due to previous major surgery (12 gastric resections, 3 resections of the colon, 1 splenectomy, 1 case of fundoplication for hiatal hernia, 1 resection of the ileum). In all cases access was by open laparoscopy to the lower right or periumbilical abdominal quadrant. When necessary, local viscerolysis was performed using various techniques with variations depending on the adhesions encountered. In all cases surgery was performed laparoscopically. In 2 cases a calculosis of the common bile duct was treated at the same time. The average duration of surgery was 70 min for cholelithiasis and 145 min for cholecysto-choledochal calculosis. On average patients were released from hospital 4 and 7 days after surgery respectively. No major or minor complications were encountered. PMID- 9411289 TI - [Critical review of treatment outcome in 54 patients with synchronous liver metastasis of colorectal cancer]. AB - Short- and long-term results of the treatment of 54 patients (12.5%) with synchronous hepatic metastases were critically reviewed by means of retrospective analysis of 431 colorectal cancer patients surgically treated over a period ranging from January 1980 to December 1989. Incidence and stage of hepatic metastases (Gennari Classification, 1984) are not significantly correlated to stage (T3), grade (G2-G3) and mucinous colorectal tumours; but they are significantly correlated to metastatic lymph nodes (p < 0.01). Actuarial survival is significantly influenced by surgery (p < 0.01) and stage of liver metastases (p < 0.05). The restriction of preoperative exclusion criteria and the simultaneous surgical treatment of primary colorectal and secondary hepatic metastases seem to be responsible for the high rates of operative mortality. PMID- 9411290 TI - [Clinico-pathological study of microcarcinoma of the thyroid]. AB - We have analysed the results of surgical treatment for microcarcinoma of the thyroid (MCT). In sixteen patient clinical and follow-up data were retrospectively evaluated during a 35.1-month follow-up. Thyroid hyperfunctional state us was present in two subjects. A single nodule was detected by echotomography in 11 patients, while multinodular diffuse goitre was revealed in 3 patients. In the last two subjects, thyroid gland appeared completely normal at ultrasonography, despite laterocervical lymph node metastases. Fine-needle aspiration biopsy was performed in 6 patients and its diagnostic accuracy was high (83,3%). MCT was classified as "incidental" in 12 patients and "occult" in the remaining 4 patients. Eight subjects underwent total thyroidectomy and 8 hemithyroidectomy plus isthmectomy. No postoperative complications were recorded. In 10 patients MCT histotype was papillar adenocarcinoma, in 5 was follicular adenocarcinoma and in the remaining case it was medullary carcinoma. Goitre was associated in 75% of the cases. Only in a patient disease progressed to death because of hematogenous metastases. In conclusion, we believe that incidental MCT is a low-grade malignancy with a benign biological behaviour. Occult MCT is a potentially lethal disease. We did not observe differences in the long-term results between different surgical treatments of MCT. PMID- 9411291 TI - [Postoperative complications after thyroid surgery. Review of the literature and personal experience]. AB - The authors make a review of the literature on the complications which occur after thyroidectomy. They report, moreover, a study carried out on 192 thyroidectomy performed between January 1992 and May 1995. The surgical technique employed foresees the systematic search for and saving of laryngeal nerves fibers and the parathyroid gland. Results were the incidence of postoperative hypocalcemia and of permanent hypocalcemia. The authors also analyse problems related to laryngeal nerves in matters of thyroid surgery. After a short introduction on surgical anatomy, attention is drawn to the "recurrent nerve risk" but also the superior laryngeal nerve too, often injured with modifications of the vocal tone and serious consequences for particular professional groups. PMID- 9411292 TI - [Spontaneous and traumatic pneumomediastinum. Analysis of 34 cases]. AB - Pneumomediastinum (spontaneous, iatrogenic and traumatic) is a relatively uncommon infrequently reported entity. The most common cause is the rupture of marginal pulmonary alveoli, allowing bubbles of air to dissect along the vascular sheaths and connective tissue planes to the mediastinum. Rupture of the trachea or thoracic traumas are other causes of pneumomediastinum. The most common presenting complaint was retrosternal pain, dyspnea, dysphagia, weakness and neck pain. Physical finding revealed: subcutaneous emphysema extended to face, chest or neck, and Hamman's sign. Chest X-ray was made in all cases and diagnosis was completed with chest CT scan and tracheoscopy. We present our series of 34 PM between January 1.1993 to July 31.1995 and discuss about etiology, diagnosis and treatment of this entity. PMID- 9411293 TI - [Diagnosis and treatment of traumatic diaphragmatic hernia with delayed presentation]. AB - AIM: To evaluate the clinical picture, diagnostic techniques and most appropriate treatment in traumatic diaphragmatic hernia with delayed presentation on the basis of personal experience and in the light of other published studies. EXPERIMENTAL DESIGN: Review of cases treated. SETTING: Patients treated in University General Surgery wards. PATIENTS: Those patients in whom diagnosis was made some time after trauma and after the acute event were selected from a group of patients with traumatic diaphragmatic hernia. SURGERY: All patients underwent surgery to reduce hernia and repair the diaphragmatic lesion. MEASUREMENTS: All clinical findings were examined together with the tests performed and the type of treatment carried out. RESULTS: The diagnosis was made between 3 months and 3 years after the injury. Three patients presented manifest symptoms of high intestinal occlusion on entry. Radiological alterations were present in standard chest X-rays in all patients and digestive tract contrast radiography was positive for the diagnosis of hernia in 3 out of 4 cases in which it was performed; a preoperative diagnosis of hernia was obtained in 4 cases. Patients were operated using a thoracotomy (3 cases) or combined laparothoracotomy access (2 cases); the diaphragmatic lesion, localised in all cases in the cupula of the left hemidiaphragm, was repaired using separate sutures in non-reabsorbable material without the use of grafts. One patient died postoperatively owing to septic complications. CONCLUSIONS: traumatic diaphragmatic hernia with delayed presentation involves severe complications that increase morbidity and operating mortality. PMID- 9411294 TI - [Paget's disease of the breast]. AB - Paget's disease of the nipple is a rare lesion that is not always associated with underlying breast cancer. The authors report three cases of Paget's disease; in two of the three patients examined, the disease became evident following the appearance of an erythematous and erosive lesion on the nipple and areola, but no clinical evidence of an underlying nodule. In both cases diagnosis was made by integrating cytological tests by apposition of the nipple with mammography. In the third case the disease became manifest following the onset of a lesion on the nipple and areola with the characteristics of chronic eczema in association with a retroareolar nodule, measuring 3 cm, with the mammographic characteristics of an infiltrating carcinoma. All three patients were treated by radical mastectomy modified according to Madden. PMID- 9411295 TI - [Follow-up of surgical biopsies in microcalcifications of the breast. Comparative analysis of patients submitted to mammography and digitalization of mammographic images]. AB - Improvements in the techniques of preoperative needle localization of nonpalpable breast lesions that have been detected at mammography, coupled with surgical biopsy of smaller volumes of breast tissue and the use of local anesthesia have produced a more aggressive attitude toward early biopsy of lesions that are suspected of malignancy. The authors report the follow-up in 92 cases, who underwent breast biopsy for microcalcifications with no palpable lesions. In 46 women the presence of microcalcifications was evaluated through a computerized instrument which allows digitalization of the image. PMID- 9411297 TI - [A case of forgotten giant goiter]. AB - The forgotten goiter is most often the consequence of the incomplete removal of a "plunging" goiter, but it can sometimes be attributed to a concomitant, unrecognised mediastinal goiter which is not connected to the thyroid. Differential diagnosis must be made with other mediastinal masses and with plunging relapses of a previously operated struma. Radiological analysis of persisting mediastinal involvement before and immediately after surgery is the only decisive means of diagnosis, but this is not always available in practice. In this paper the authors report a case of considerable size observed in a series of 346 mediastinal goiters operated between 1967 and 1994. They examine the pathogenetic aspects and the nosological, diagnostic and therapeutic problems related to forgotten goiter, and lastly they list the recommendations that several surgeons have made in an attempt to reduce the incidence. In conclusion, the systematic use of CAT or NMR in the diagnosis of mediastinal opacity may help to reduce the risk of forgetting glandular residue in the mediastinum. PMID- 9411296 TI - [Totally implantable venous access systems. Analysis of complications]. AB - Totally implantable central venous access devices (Port-a-Cath, PaC) allow better treatment of cancer patients, with safe administration of chemotherapeutic agents, and are well accepted by the patients. The aim of the present paper is to analyze the complications of the different implant techniques on the basis of a personal experience of 92 central venous access devices. MATERIAL AND METHODS: A total of 92 PaC (Port-a-Cath, Pharmacia: Celsite Braun) have been implanted in 88 patients between August 1992 and June 1995 for cancer treatment. Age ranged between 19 and 79 years (median 52 years), 56 were male and 32 women. PaC have been implanted by percutaneous cannulation of the subclavian vein, with Seldinger technique, in 34 cases; by venous cutdown respectively on the cephalic vein in 46 cases, the jugular vein in 7 cases, the basilar vein in 4 and the saphenous vein in 1 case. Four patients experienced a double implant. In 84 cases the implant was done under local anesthesia, while in 8 required general anesthesia, during operation for the primary neoplasm. RESULTS: A total of 7 complications were experienced (7.6%, 7/92): 4 sepsis and 3 mechanical. No cases of pnx were observed. Sepsis occurred after 29, 45, 64, 401 days of implantation respectively, and culture demonstrated S. aureus in 2 cases, and E. coli and Klebsiella oxytoca in 1 case each. Mechanical complication comprehends 2 cases of catheter dislodgement and 1 case of port rotation. No complications were noticed in case of implant during surgery for primary cancer (8 cases). In 7 cases the procedure has been converted from cephalic vein cutdown to percutaneous cannulation of the subclavian vein due to anatomic reasons (13.2%, 7/53). Five PaC have been explanted for complications. DISCUSSION: On the basis of the personal experience we think that PaC are of easy implant, with few complications and of good acceptance from the patients. We prefer venous cutdown on cephalic vein as implant technique because of avoidance of pnx or bleeding complications. Percutaneous puncture of subclavian vein is useful for implantation during major surgery, because less time consuming, and in case of anatomical anomalies fo the cephalic vein. Basilic vein cutdown has been utilized exclusively for esthetic reason in young people, to avoid the scar in the upper thoracic region. Alternative implant techniques has been employed in special conditions, such as catheter position in the inferior v.cava, or early in our experience (internal jugular vein). A total of 7 complication have been reported (7.6%), 4 sepsis and 3 mechanical (2 dislodgement, 1 rotation). Sepsis were not related to implant technique, presenting on day 29, 45, 64 and 401 respectively; all required the explant of the PaC as a treatment. Mechanical complications are related to surgical technique; all required re-exploration with 1 explant and 2 reposition of the PaC. In PaC positioning during surgery for primary cancer (8 cases) no morbidity has been reported. All but the 5 PaC explanted were functioning until patient's need; maximum length reported is 42 months. PMID- 9411298 TI - [Solitary cyst of the liver]. AB - The rareness of the illness and the infrequency of infectious complications have led the authors to report a case which has come to their attention. After a brief exam of the etiopathogenesis and pathologic anatomy of these hepatic forms of cyst and the following considerations on their clinical aspects, which range from complete lack of symptoms to the compression of nearby organs (gall ducts, stomach, kidney) due to an increase in their volume, to occurrence of complications (rupture, haemorrhage, suppuration), the authors evaluate the therapeutic problem. Emptying and internal drainage being nowadays discarded and everyone agree on the necessity of excising the whole cystic wall with the minimum loss of hepatic parenchyma, the most indicated operation is total cystectomy. As such an operation is not always feasible because of the adherences of the cyst to the hepatic parenchyma; partial cystectomy can be practiced, leaving in situ part of the cystic wall. If the cyst is complicated by rupture, haemorrhage or suppuration, typical or atypical hepatic resection is recommended. The reported case concerns a patient in good general conditions, bearer of a large left hepatic lobe cyst, complicated by suppuration, with normal liver function, who has been submitted to ablation of the third hepatic segment according to the Ton That Tung transparenchymal technique. Finally the authors point out the possibility of a new conservative therapeutic approach, practiced in a few centres, consisting in echo or CT guided percutaneous needle suction and sclerosis of the cyst with 95% pure alcohol. PMID- 9411299 TI - [Acute pre-hepatic portal thrombosis: diagnosis and therapy]. AB - Portal vein thrombosis is a rare pathology. The etiopathogenetic causes that most frequently lead to this pathology are myeloproliferative syndromes. The authors present a case of acute pre-hepatic portal vein thrombosis and discuss its etiopathogenesis, diagnosis and therapy. PMID- 9411300 TI - [Gastric schwannoma]. AB - A rare case of gastric schwannoma is reported. A 36-year-old woman whose endoscopy showed a mucosa tumor, biopsy findings were suggestive for leiomyoma. Diagnosis was made only with postoperative histology that revealed the benign schwannoma. Immunohistochemical staining showed positivity for S-100 protein and the neuron specific enolase, schwannoma is often associated with Von Recklinghausen's disease. Surgical resection is recommended as their malignant potentiality. Gastric and small intestine localizations are often with bleeding. They also represent almost the 24% of all gastrointestinal stromal tumors (GIST) and the 4% of all primary retroperitoneal tumors. PMID- 9411301 TI - [Hemorrhagic pseudocysts of the spleen]. AB - After a short introduction regarding the physiopathology of secondary spleen cysts, the authors describe a personally observed case of voluminous hemorrhagic pseudocyst. They underline the rarity of the orientation towards radical surgery, having in mind the post-splenectomy risks determined by deficiency of important spleen functions. The authors, with respect to this, underline the importance of reimplanting splenic tissue fragment into omentum, promoting to use, in selected cases, this treatment that needs, however, further clinical findings. PMID- 9411302 TI - [Intestinal angiodysplasia: diagnosis and therapy]. AB - On the basis of two personal cases of angiodysplasia authors describe this rare pathology and its relative diagnostic difficulties. On the grounds of their experience and the international literature on this pathology, authors considered the angiography as the selected method for the diagnosis of intestinal angiodysplasia, and they consider surgical therapy as the best curative method for this pathology. PMID- 9411304 TI - [Problems of differential diagnosis of lymphoma and celiac disease. A case report]. AB - An association between celiac disease and non-Hodgkin's lymphoma of the small intestine has been recognized for many years. Coeliac disease is characterized by an enteropathy sensitive to gluten, malabsorption of food and partial or total villous atrophy. Also malignant lymphoma may present with malabsorption and mucosal lesion similar to that found in coeliac patients. The diagnosis of lymphoma in coeliac patients can be extremely difficult because the presenting symptoms and histological lesion are similar, but the presence of a cluster of symptoms such as abdominal pain malabsorption, weight loss in patients older than 40 years with a history of poorly responsive coeliac disease should raise a suspicion of malignancy. We present a case of 55 year-old man with malignant lymphoma and coeliac disease surgically treated in our Institute for intestinal obstruction. PMID- 9411303 TI - [Neutropenic colitis. A case report]. AB - The authors take the opportunity, on the basis of a clinical case examined, to revise an uncommon disease: neutropenic colitis. In the presence of an acute abdominal pain and tenderness localized at right lower quadrant, in a patient affected by hematologic neoplasm or by a solid tumor, at the end of a chemotherapeutic treatment with absolute residual neutropenia, we would be strongly oriented to neutropenic colitis. The treatment according to the primary disease and to the patient's poor conditions, will be where possible conservative, reserving the surgical approach only for those cases of absolute necessity. PMID- 9411305 TI - [Intestinal obstruction caused by paracecal hernia. 3 case reports and review of the literature]. AB - The authors report three rare cases of intestinal obstruction due to paracecal hernia observed in 533 small bowel obstructions operated between January 1982 and December 1994 (0.6%). In our experience, all the cases occurred in old female patients. Less than 150 cases are reported in the literature. The authors examine paracecal hernia embryologic aspects to explain its pathogenesis: the rotation of primary intestinal loop determines final intestinal rapports. Preoperative diagnosis is very difficult. Transitory symptoms may appear months or years before intestinal obstruction; these occurrences suggest that internal hernias, like external ones, may spontaneously reduce. Straight abdominal radiographies, performed in all cases here described, demonstrate small bowel levels. There is disproportion between important subjective symptoms and objective finding of a large round bump localized in the right iliac fossa. Authors underline the opportunity of a promptly performed operation: in all our cases we released the incarcerated intestinal loops and sutured hernial foramen. One exitus occurred, on the 4th postoperative day, probably due to pulmonary embolism. PMID- 9411307 TI - [Sarcomatoid renal carcinoma. A case report]. AB - Renal sarcomas are rare tumors. Prognosis is overall dismal. Adjuvant therapies should follow radical nephrectomy but no standardized regimen has been at present defined. We report a case of a patient affected by a sarcomatoid renal tumor to detect the best therapeutic approach to this rare tumor. PMID- 9411306 TI - [A case of ileal perforation caused by Crohn's disease]. AB - A case of acute perforation of the small bowel in a 32-year-old man with Crohn's disease not previously diagnosed is presented. The patient underwent an emergency bowel resection and anastomosis and was put on 5-ASA for the following 4 years. No clinical or endoscopic recurrences appeared during this period. In order to avoid both functional disorders and recurrences, a surgical treatment based on resection and anastomosis in minor risk cases, i.e. in patients without abscess and/or concurrent steroid therapy, and on resections limited to grossly involved segments of bowel is advocated. PMID- 9411308 TI - [Klippel-Trenaunay syndrome: clinical and surgical aspects]. AB - The Klippel-Trenaunay syndrome is a rare syndrome of uncertain etiology. The characteristic elements are flat angiomatosis, hypertrophy of soft tissue and bone tissue and alterations of the venous system, with the exclusion of hemodynamically significant arteriovenous fistulae. The authors report a clinical case and review the international literature. Treatment is conservative in the majority of cases; surgery is reserved for patients with disabling morphological and functional alterations. PMID- 9411309 TI - [Platelet activating factor (PAF) and kidney failure progression]. AB - Many mediators of phlogosis may play an important role in renal glomerular pathology. PAF seems to have a prominent role. The aim of this study is the evaluation of the mechanism of action by which PAF may contribute to renal glomerular damage. PMID- 9411310 TI - [Review of 87 cases of scoliosis surgically treated]. AB - BACKGROUND: The authors compared the results and complications in surgical treatment of idiopathic scoliosis with Harrington's rod instrumentation with subtrasversal wires in dorsal treat. METHODS: A research on 87 cases operated on for idiopathic scoliosis from 1987 to 1995 is carried out. The 87 cases include 65 females and 25 males, 16 years old in average (range, 11 to 30). The curvature in Cobb's degrees and rotation of vertebrae with Raimondi's method on radiographs take just before, 15 days later and an year later on operation was measured. The patients have been divided into two groups: the first of 77 patients operated with Harrington's rod instrumentation; the second of 10 patients operated with Harrington's rod instrumentation and subtrasversal wires. RESULTS: In a general analysis without taking in to consideration the type and the seriousness of curvature it was obtained a better correction and derotation of vertebrae in patients of second group. The same group with wires had obtained a better correction and derotation of vertebrae in dorsal scoliosis from 40 degrees to 60 degrees and in the double scoliosis, while the first group obtained better results in dorsal scoliosis from 60 degrees to 80 degrees and in derotation of vertebrae on dorsal treat of double scoliosis. One case of pseudarthrosis in every group was observed. Any neurological complication were observed. CONCLUSIONS: The conclusions is drawn that the application of subtrasversal wire improves the Harrington's technique for the correction and derotation of dorsal and double scoliosis without neurological complications sometimes present with subliminar wires. PMID- 9411311 TI - [Cardiovascular disease and omega-3 fatty acids]. AB - Fish oil is rich in the long chain omega-3 (omega-3) polyinsaturated fatty acids (PUFA), Pioneering studies of Dyerberg and Bang primarily originate interests in this way. The low incidence of acute myocardial infarction they verified within the Greenland Eskimos suggested that a high dietary omega-3 PUFA intake due to marine food might protect against coronary heart disease. They showed that the Eskimos had a beneficial lipid pattern and that their balance between pro aggregatory thromboxanes and anti-aggregatory prostacyclins was shifted towards an anti-thrombotic state. The two major omega-3 fatty acids are decosapentaenoic acid (EPA C 20:5, omega 3), with five double bonds, and docosahexaenoic acid (DHA C 22:6, omega 3), with six double bonds. These fatty acids' significant effects include reduction of plasma triglycerides and lipoprotein levels as well as of platelets thrombogenicity in the microcirculation, which is due to effects on the mediators production derived from arachidonic acid (prostaglandins and leucotrienes), meddling in inflammatory and immune cell function, retarded atherosclerosis development. Experimental studies of atherogenesis and arterial thrombogenesis support the hypothesis that dietary omega-3 PUFA intake may play a leading role in primary or secondary prevention of coronary heart disease. PMID- 9411312 TI - [Endocrine diseases and male infertility]. AB - Male infertility is secretory or excretory. The excretory form lies outside the endocrinologist's evaluation. As regards the secretory form, infertility practically identifies itself with hypogonadisms, both primitive, congenital or acquired, that spring from testicular alterations, and secondary to that result from gonadotropic deficiency however derived. The various diseases and the large number of events responsible for primitive or secondary male hypogonadism and so cause of infertility, are minutely examined. Peripheral hypogonadism forms are mentioned in brief too. During diagnostic evaluation, anamnesis and objective examination are the first step. Afterwards hormonal basal dosages supply useful information in case of severe oligospermia or azoospermia. Particularly FSH plasma dosage values the tubular function and this hormone is the most significant clinical and prognostic parameter. LH plasma dosage is less useful. FSH and LH levels allow us to distinguish hypergonadotropic hypogonadism from the hypogonadotropic type. Testosterone dosage is also important. Prolactin measurement is useful too and likewise, in particular cases, other hormone measures. Dynamic tests are also mentioned. As a therapeutic guideline, treatment with androgens is exclusively substitutive in primitive hypogonadism, however of no avail on spermatogenesis; in cases of secondary hypogonadism, gonadotropins give satisfactory results; pulsatile Gn-RH use is not easy anyway; clomifen and tamoxifen don't seem of particular advantage; in hyperprolactinemia, dopaminergic drugs improve seminal parameters. PMID- 9411313 TI - [Guidelines for the treatment of acute asthma. A critical retrospective analysis]. AB - Acute asthma attacks are frequently observed in common medical practice. However a unified and scientifically proven directive which advises objectively on the best management strategy for patients with acute asthma attacks is still lacking. In the present review we have accurately assessed various important points addressed in the recent international guidelines for acute asthma. This work has enabled us to derive precise and documented suggestions of practical importance which may be useful to the physician for a correct and judicious approach to the management of those patients with acute asthma attacks. PMID- 9411314 TI - [Plasma protein adsorption on variously treated +polybutylene terephthalate]. AB - BACKGROUND: The aim of the research was the evaluation of plasmatic protein adsorption on untreated polybutylene terephthalate, corona treated polybutylene terephthalate and polyvinylacetate coated corona treated polybutylene terephthalate. METHODS: Total proteins, albumin, immunoglobulins, fibrinogen, insulin and ostecalcin were determined on plasma after contact with these polymers. RESULTS: Unsignificant variations were observed for the assayed proteins. CONCLUSIONS: The conclusions are drawn that the tested materials do not adsorbe significantly the most important plasmatic proteins. PMID- 9411315 TI - [Superovulation induced by the administration of PMSG to mustelids]. AB - In this paper, we have attempted to use the technique of superovulation in mustelids. Ermine (Mustela erminea) and several species of ferret (Mustela eversmanni, Mustela putorius, Mustela putorius furo [correction of hamster (Putorius eversmanni, Putorius putorius, Putorius putorius furo)] have been used as experimental models. We have conducted a detailed study of superovulation and early embryonic development in animals of these species treated with PMSG [correction of GPMS]. We have shown that superovulatory response is produced only in the right, but not in the left ovary of hormonally treated animals. Embryonic development in hormonally stimulated animals was accelerated as compared with animals that were reproducing under natural conditions. Perspectives and limitations in the use of hormonal treatment with the purpose to induce superovulation in mammals are discussed. PMID- 9411317 TI - [Ultraweak emissions from chicken eggs and embryos: the nonadditive interaction of 2 emitters and stable nonequilibrium]. AB - Ultraweak emission of the optic range from developing, unfertilized, and dead chicken eggs and their components (isolated blastoderm and embryo, entire yolk, white, and shell) was measured at the normal and abnormal temperatures of incubation using a photomultiplier tube. Two sources of ultraweak emission were found: blastoderm and yolk from the fertilized eggs until day 4 of incubation and shell from all eggs, including the unfertilized and dead ones. Emission from the former source was weaker and almost light independent, was recorded only at the temperature of incubation, and had a wavelength of no more than 3000 nm. Emission from the latter source was distinctly light dependent, less temperature dependent, and had a wavelength of 600-800 nm. Ultraweak emission of the entire fertilized eggs was not equal to those of their components: at the early stages of incubation, the internal components of the eggs mostly stimulated ultraweak emission of the shell, while from day 9 of incubation on, they inhibited or absorbed the latter. Absorption and emission of photons by the shell increased as the development proceeded. As compared to the unfertilized and dead eggs, emission from the entire developing egg was characterized by a greater sensitivity to temperature gradients and resistance against small temperature fluctuations in the optimal temperature range. Analysis of the curves of afterillumination decrement of ultraweak emission suggests a significant coherence of emission from the entire developing egg and whole shell, rather than from the unfertilized and dead eggs and shell fragments. PMID- 9411316 TI - [The role of serotonin in the embryogenesis of normal and irradiated mouse embryos]. AB - We have examined the role of serotonin, a neurotransmitter, in the development of normal embryos and embryos irradiated in utero at different stages of morpho- and organogenesis. We have found that serotonin is capable of attenuating radiation induced disturbances of development; serotonin therapeutic action varied depending on the age of the embryo, severity of the original radiation damage, and the time that serotonin was administered after irradiation. All data provide evidence that the window of serotonin's therapeutic effect is strictly timed to a certain stage of prenatal ontogenesis, specifically, to neurogenesis. This conclusion is in agreement with the view that serotonin serves as a morphogenetic signal for the developing nervous system, the state of which is important for the viability and functionality of fetuses in the normal state and after exposure to ionizing irradiation. PMID- 9411318 TI - [Cytokeratin expression in the pre- and postnatal ontogeny of the rat liver]. AB - Expression of cytokeratins nos. 7 and 19 has been examined during pre- and postnatal ontogenesis in rat liver. Cells expressing cytokeratin no. 19 appeared around large incoming vessels in the liver at days 17-18 of gestation. From day 20 of gestation and during the first week of postnatal ontogenesis, cholangiocytes and periportal hepatocytes could be stained with antibodies against cytokeratin no. 19. The expression of cytokeratin no. 7 begins later than cytokeratin no. 19, and it is present only in cholangiocytes, throughout pre- and postnatal ontogenesis. Since pre- and postnatal hepatocytes are capable of expressing cytokeratin no. 19, we discuss possible use of this marker to study cytodifferentiation of epithelial cell lines in rat liver. PMID- 9411319 TI - A novel, simple organotypic culture method to study the organ of Corti from the neonatal gerbil. AB - An original, simple organotypic culture method was developed to grow the organ of Corti from the neonatal gerbil on the bottom of a Petri dish. In comparison with the commonly used Maximov slide assembly method, this method is easier, less time consuming, and more economic. Our results in this study using fluorescent live/dead viability assay and fluorescein-conjugated antineurofilament antibodies show that the cultured organ of Corti and spiral ganglion cells not only survived for at least 14 days but also maintained their basic organization and normal development in vitro. Therefore, our method can serve as a reliable and easier alternative to the traditional techniques for studying the development as well as other physiological properties of the cultured organ of Corti. PMID- 9411320 TI - [The place of pelvic adhesions in gynecology. Possibilities of prevention]. AB - The authors review the recent international literature dealing with the role of pelvic adhesions in gynaecologic surgery. They point out that pelvic adhesions can be detected via laparatomy or laparoscopy in every third patient suffering from chronic pelvic pain, whereas abdominal adhesions revealed rarely correlate with anamnestic chronic pelvic pain. Correlation of pelvic pain and adhesions is neither consequent nor mutual. Lacking more detailed information on the pathomechanism of pain, the role of the vegetative innervation of the visceral and parietal peritoneum and of the excitation of the nerve fibers in the neoformed-and vascularized adhesions can only be postulated. According to the attractive but not well established theory extensive adhesions can directly provoke pain through fixing the pelvic organs hampering their motility and mobility. It is generally accepted, that lysis of adhesions in symptomatic patients can resolve or reduce the complaints. However the duration of the beneficial effect and its transitoric or definitive nature cannot be predicted in advance. Subsequently the authors review the adjuvant methods complementary to microsurgical techniques possibly preventing the formation and neoformation of pelvic adhesions. Despite of general acceptance and widespread usage, the effectiveness of crystalloids, macromolecular solutions, intraperitoneal heparin and steroids in the prevention of adhesions cannot be scientifically supported. The encouraging results already achieved by mechanical barriers (intercede, Gore Tex) can hopefully be surpassed by biodegradable barriers actually studied in animal models. PMID- 9411321 TI - [Role of aldosterone in potassium secretion in chronic renal failure associated with hypertension (transtubular potassium gradient)]. AB - Aldosterone protects against hyperkalemia in disorders with reduced number of over-working nephrons. It is not clear however, whether diminished aldosterone production or aldosterone resistance is responsible for the hyperkalemia in chronic renal failure. The importance of this question is underlined by the fact that antihypertensive drugs often reduce aldosterone level. Therefore we investigated the distal tubular potassium "driving force" (transtubular potassium gradient) in 9 patients with chronic renal failure accompanied by hypertension and compared it to the results obtained in 10 healthy persons. Glomerular filtration rate was 15.77 +/- 4.88 ml/min in the chronic renal patients. Serum potassium was normal in 3 of 9 patients, the average 4.88 +/- 0.16 mmol/l, higher than in healthy persons 4.27 +/- 0.09 mmol/l (p < 0.001). Nevertheless the transtubular potassium gradient was lower in patients (3.52 +/- 0.32) comparing to healthy persons (7.25 +/- 0.57, p < 0.001). The patients' plasma aldosterone was much higher than normal. The reduced tubular potassium secretion-decreased "driving force" --suggested to aldosterone resistance. We conclude that the renal tubules' aldosterone resistance is not an exceptional but rather frequent feature in chronic renal failure which should be considered when administering antihypertensive drugs. PMID- 9411322 TI - [Early diagnosis and management of femur head necrosis in young adult age]. AB - There are a lot factors in the genesis of the ischemic femoral head necrosis. In the early stage the intraosseous pressure increases without any clinical or radiological sign. The disease is often bilateral. The early diagnosis (NMR) is essential in the treatment. In the case of a severe femoral head necrosis the radiological investigation of the other side is obligatory, in order to diagnose the early stage of the disease and to prevent the progression of the collapse with core decompression. The object of the adequate treatment of the collapsed femoral head is to release the pain and to avoid the total destruction of the femoral head. The untreated femoral head necrosis results in a painful, arthritic hip, where spontaneous regeneration is limited, and takes 8-10 years. With osteotomy and revascularisation the results are good, and the regeneration takes about 2-3 years. As a result of this treatment, the patients have a moderately limited function, and minimal limping. Rehabilitation is possible into the original profession, or into a somewhat easier job. Remodellation of the femoral head in younger patients following the revascularisation procedure has been observed in our experience in the last few years. PMID- 9411323 TI - [Therapeutic importance of the diagnosis of Kallmann syndrome]. AB - Hyposmia with hypogonadotropic hypogonadism was diagnosed as Kallmann syndrome in a 24 years old dizygotic female twin. This syndrome indicates the importance of smell in the sexual development through the progenitor cells in the olfactory placode because luteinizing-hormone-releasing hormone (LHRH) secreting cells of hypothalamus arise from these cells. In addition, substitution therapy may be successful in the treatment of the lack of secondary sex traits and primary amenorrhoea as the presented case demonstrated. PMID- 9411325 TI - [Remembering Janos Ertl, M. D. (1880-1951)]. PMID- 9411324 TI - [A case of unusual complication of diaphragmatic herniation of transverse colon following transhiatal esophagectomy]. AB - The authors report a rare complication observed after transhiatal esophagus extirpation performed for esophageal cancer. In this case the transverse colon herniated into the pleural cavity through the esophageal hiatus. Herniation completed on the 6.th. postoperative day and caused mechanical ileus. In the first few postoperative days, radiology pointed to a basal pulmonal infiltrate, later it resembled relaxation of the diaphragm, which was rather misleading. Recognition of the real situation was possible only on the 6.th.postoperative day. The patient was reoperated and the pathological state could be reversed. The esophageal hiatus was reconstructed. In the opinion of the authors the complication may have developed partly due to the opening of the left pleural cavity in the course of extrathoracal esophagectomy, and partly to the fact that the spleen was removed during operation. The outcome after reoperation was uneventful. The authors consider this as a rather rare and instructive case. PMID- 9411326 TI - [History of the Radiology Section of the Hungarian Society of Gastroenterology]. PMID- 9411328 TI - [Combined injection/endoscopic sclerotherapy of upper gastrointestinal peptic ulcer hemorrhage]. AB - In the last 8 years period 369 patients with acute upper gastrointestinal bleeding were admitted to the Department of Internal Medicine, District Hospital Siofok. All patients were treated by combined injection sclerotherapy during the urgent endoscopy. The sclerotizing solution contained at the first step ethamsylate, calcium gluconate, epinephrin, hypertonic saline solution, and the second step 1% aethoxysklerol. At he time of urgent endoscopy 77% of the patients had acute bleeding (Forrest I.a., I.b.) and 23% of them showed stigmata of fresh bleeding (Forrest II.a., II.b.). Primary endoscopic hemostasis was achieved in all patients. In 7.9% of patients (n = 29) developed recurrent bleeding during the observation period, who were resclerotized. Twenty of them (5.4%) were treated by elective surgery. No hemorrhagic or sclerotizing therapy associated mortality occurred. Five out of 20 operated patients (1.4% of all sclerotized cases) had died due to the serious complications of their chronic liver disease. Patients with succesful initial endoscopic hemostasis were treated conservatively (proton pump inhibitors, H-2 blockers, antacids, sucralfate, lactulose) and underwent control endoscopy after 24 hours. Using this sclerotizing method in the treatment of acute upper gastrointestinal bleedings the number of acute surgery and mortality decreased significantly. It is supposed, that the locally administered ethamsylate, calcium gluconate, epinephrin, hypertonic saline solution and aethoxysklerol containing sklerotizing solution might play a favorable role in the successful control of acute bleeding ulcers. PMID- 9411327 TI - [AT1 angiotensin receptor inhibition as a new therapeutic possibility]. AB - The octapeptide hormone, angiotensin II, binds to two major subtypes of cell surface receptors: the AT1 and the AT2 angiotensin receptors. The important physiological and pathophysiological effects of angiotensin II on cardiovascular regulation and salt-water balance are mediated by the AT1 receptor subtype. As a consequence of the outstanding clinical success of angiotensin-converting enzyme inhibitors, the appearance of AT1 receptor inhibitors in the therapy of hypertension and other cardiovascular diseases was preceded with great expectations. The available experimental and clinical data indicate that the first AT1 receptor inhibitor, losartan, has the same therapeutic potential as angiotensin-converting enzyme inhibitors, but it does not evoke the angiotensin independent side-effects of ACE inhibitors, such as dry cough or angioedema. The physiological importance and the biochemical, molecular biological and pharmacological properties of AT1 and AT2 receptors are reviewed in this paper, and a summary of the available clinical data is presented. PMID- 9411329 TI - [Possibilities of the use of 3-dimensional ultrasonography in gynecology and obstetrics]. AB - The authors report their experiences with 3 dimensional ultrasound technique applied in cases of 286 obstetrical and 35 gynecological examinations. Combison 530, 3 D ultrasound equipment was used. Pathological signs were found in 14 cases. The introduction of this technique has been increasing the diagnostic accuracy of prenatal and gynecological ultrasound examinations, and some malformations, that could have never been seen by "traditional" 2 D methods, can be visualized as well. PMID- 9411330 TI - [Clinical experiences with the analgesic effects of citalopram]. AB - Antidepressive drugs influencing the serotonin metabolism have shown some efficacy in treatment of generalised pain syndrome. The aim of the present study was to observe the analgesic effect of citalopram < 1-[3-(dimetilamino)propil]-1 (p-florofenil)-5-ftal+ ++ankarbonitin > in 20 non depressive patients with chronic pain syndrome (fibromyalgy and/or radicular pain). On the base of a short time (6 weeks) observation a limited clinical effect was observed, however, the Seropram seems to have some benefits in the treatment of chronic pain. PMID- 9411331 TI - [Artificial nutrition of patients with toxic epidermal necrolysis (Lyell syndrome)]. AB - The authors report the case of a 30-year-old man treated with toxic epidermal necrolysis. Toxic epidermal necrolysis was due to anticonvulsive drug treatment. The patient was admitted with denudated skin surface similar to second-degree burn that covered 90 per cent of the patient's body surface. The patient was isolated and treated, receiving sterile wound care, broad spectrum antibiotic and corticosteroid. Total parenteral nutrition was instituted until the 5th day of care because the patient was unable to take normal food. The energy intake reached 146 kJ/kg bodyweight containing 4 g/kg bodyweight carbohydrates and 2 g/kg bodyweight fat emulsion supplemented with 10-15 g of nitrogen per day. The enteral nutrition was commenced gradually with decreasing parenteral nutrition. The nutritive solutions were supplemented with ions, vitamins and trace elements. The patient left the intensive care unit after 23 days of care. The toxic epidermal necrolysis is a life-threatening dermatological disease and should be treated at intensive care unit. The early recognition of the disease, the intensive care and nutritional therapy may improve the survival of patients with toxic epidermal necrolysis. PMID- 9411332 TI - [Molecular genetic study of type I autosomal dominant polycystic kidney disease]. PMID- 9411333 TI - [Prevalence of vertebral deformity in Hungary: the European Vertebral Osteoporosis Study]. AB - Prevalence and distribution of vertebral deformity regarded as classic marker of osteoporosis are reported on the basis of the results of the first Hungarian cross-sectional population-based survey. The clinical-epidemiological project is conducted by the National Institute of Rheumatology and Physiotherapy in Hungary in the frame of the European Vertebral Osteoporosis Study which comprises 19 countries and 36 centres. A random sample of 324 females and 300 males aged 50 years and over stratified in 5-year age bands was recruited. Lateral spinal X rays were evaluated centrally by two different morphometric methods. Based on the McCloskey-Kanis criteria the prevalence of all deformities was 16.7% in females and 18.7% in males. These values from Hungary have been considered high compared to the prevalence rates of other European countries, approaching, and in males exceeding, the highest rates gained from Sweden. In females the prevalence increased with age, men aged 50-59 years and women aged 75 years and over had higher prevalence of deformity compared to the other sex. In females vertebra thoracic 11, in males that of lumbar 3 were mostly affected, wedge deformities occurred most frequently. Extrapolating the results we can conclude that vertebral deformity found in both sexes similarly, affects approximately 500,000 individuals in Hungary along with the potential deterioration of quality of life. This enormous burden requires the consistent fulfillment of the National Osteoporosis Program and other preventive-curative strategies. PMID- 9411334 TI - [Glycoprotein hormone alpha-subunit secretion in non-functioning pituitary adenomas]. AB - The aim of the present study was to investigate the prevalence of elevated free glycoprotein hormone alpha-subunit in different pituitary adenomas, to establish the diagnostic value of the basal and stimulated free alpha-subunit secretion in non-functioning adenomas. Serum basal levels of alpha-subunit were increased in 1 of 22 untreated, in 1 of 16 operated patients with non-functioning adenoma, in 6 of 28 untreated, in 1 of 7 operated patients with acromegaly, in 0 of 5 untreated prolactinomas and in 0 of 1 untreated gonadotrop adenoma. Overall free alpha subunit levels were increased in 9 of 79 cases (11.4%). In 6 of 9 patients with untreated non-functioning adenoma thyrotrop hormone releasing hormone caused an abnormal--paradox--elevation of serum alpha-subunit. These data indicate that measurement of basal and stimulated alpha-subunit is of relatively poor value in the diagnosis of non-functioning pituitary adenomas. The transsphenoidal surgery did not resulted in a change of alpha-subunit secretion neither in patients with non-functioning adenoma nor with acromegaly. The present data confirm the view that non-functioning pituitary adenomas are not homogeneous since this subset of tumors includes adenomas that either do not secrete measurable amounts of free alpha-subunit or produce normal or supranormal amounts of subunits as consequence of still undefined biosynthetic abnormalities. PMID- 9411335 TI - [Effect of betaxolol on blood pressure and heart rate in mild to moderate hypertension]. AB - The efficacy and safety of daily 20 mg betaxolol monotherapy was investigated in mild-moderate essential hypertension in a four week long, open label, single blind trial (with a placebo run-in). Twenty one patients of both sexes were enrolled. The systolic blood pressure in the supine position decreased from 158 to 142 mmHg, the diastolic blood pressure from 101 to 89 mmHg. The mean systolic values of the 24 hours ambulatory blood pressure monitoring decreased from 136 to 126 mmHg, the mean diastolic values from 87 to 80 mmHg. All decreases in blood pressure were significant. The reduction of the heart rate (80/min vs 63/min) was also significant. The decrease in blood pressure during daytime was significant, during night it was moderate. The blood pressure- and heart rate reducing effect of betaxolol was detectable however in the second half of the night, before wake up. No side effect was recorded. PMID- 9411336 TI - [Thin glomerular membrane nephropathy. Clinico-pathological observations]. AB - In two nephrology centres between 1983 and 1993 among 1545 kidney biopsies 34 cases of thin basement membrane nephropathy have been diagnosed. All patients had a varying degree of microscopic dysmorph haematuria, occasional slight proteinuria--except two nephrotic children; and normal blood pressure with one exception. 5 children and 7 adults experienced repeated bouts of macroscopic haematuria mainly after exercise or upper respiratory tract infection, one child after tonsillectomy. All patients had normal renal function and retained it during the follow-up period (mean 61 months, 3 months to 22 years), except a 46 year old patient, who was found to have the joint occurrence of light chain gammopathy and hypertension. Seven patients had positive family history for microscopic haematuria, in four family members of three patients renal biopsy disclosed mesangioproliferative glomerulonephritis with thin GBM segments. As a cut off value for thin basement membrane nephropathy we considered 264 nm. The morphometric analysis of the electron micrographs revealed a mean thickness of 210 nm. No differences in basement membrane thickness were measured regarding gender, age or the presence of macroscopic haematuria. The thin basement membrane is considered to be the pathological basis and predisposing alteration leading to haematuria. PMID- 9411337 TI - [Peritoneal hemorrhage caused by trophoblast tissue implanted on the peritoneum 6 weeks following induced abortion]. AB - A case of heavy intraperitoneal bleeding from perimetrical implanted trophoblast tissue on the uterus was reported following artificial abortion. The six weeks previously terminated pregnancy was verified by transvaginal ultrasound examination, the unrecognised simultaneous extrauterine pregnancy was excluded. The mechanism of peritoneal implantation of trophoblast tissue is unknown. The authors reported the successful conservative treatment and analyzed possible origin of this rare localisation of trophoblast tissue. PMID- 9411338 TI - [On the 75th anniversary of the death of Jozsef Arkovy]. PMID- 9411339 TI - [Pathography of Winston Churchill (1874-1965)]. PMID- 9411340 TI - [Brief history of terminology of medical literature]. PMID- 9411341 TI - [New developments and trends in adenovirus research]. AB - Adenovirus infections have been associated with a variety of disorders of the respiratory, ocular, gastrointestinal and urogenital systems. In the last fifteen years, special attention has been given to the adenoviral infections in immunocompromised host, transplant recipients, cancer patients and AIDS patients. Adenoviruses in these patients can cause severe, often generalized illness, with high fatality rate. Data suggest a specific role for adenoviruses in AIDS pathogenesis. From AIDS patients many new and intermediate adenovirus serotypes could be isolated. Considerable effort should be devoted to this area of research in the coming years to understand the molecular mechanism of the interaction between AIDS and adenoviruses. In the last few years recombinant adenoviruses have been widely used as gene delivery vectors in experiments both with curative and preventive purposes. Adenovirus vectors have been used in the experimental gene therapy of genetic disorders, of immuno- and molecular therapy of a variety of cancers. The combination of recombinant adenovirus technology with chemotherapy (pro drug system) seems to be promising, as well as the specific destruction of tumor cells with modified, not recombinant adenoviruses. An the other hand, recombinant adenoviruses appear to be attractive candidates for vaccination against the infectious diseases, too. The current tendency is the construction of second-generation vectors which serve better the different purposes. Potential improvements could be the construction of targated vector by the modification of viral cell-tropism and the suppression of the immune response of the host organism directed against the vector and the vector virus infected cells. PMID- 9411342 TI - [Striking increase in mortality from venous diseases in Hungary]. AB - The statistical data of the World Health Organisation were studied from 1970 to 1990 to evaluate changes in mortality caused by venous diseases in 9 European countries and in the USA. The data were analysed in process and mortality rates in Hungary and in other countries were compared. In 1970 data based on general population of Hungary indicated medium incidence of mortality due to vein pathology (10.8/100,000) compared to the economically more developed countries. Unfortunately by 1990 mortality due to venous diseases almost had doubled (19.8/100,000). During this 20 years period mortality rates caused by venous diseases gradually decreased in the USA and in most European countries, e.g. by 80% in Austria and by 70% in Finland. Even the relatively high mortality caused by venous diseases in Switzerland (10/100,000) comprises only 50% of that demonstrated in Hungary. While the significance of modern thromboprophylaxis was recognised in the Western European countries and the USA in the early seventies, a delay of the introduction of prophylaxis with low-dose heparin was observed in Hungary. Socio-economic and nutritional causes could have been contributed to the high mortality rates caused by venous diseases as well. Further epidemiologic investigations focusing on venous morbidity factors and wide scale application of organised preventive programme are suggested. PMID- 9411343 TI - [Lower bone density (osteopenia) in adolescent girls with oligomenorrhea and secondary amenorrhea]. AB - Occurrence of reduced BMD among adolescent girls and young women due to certain specific oligomenorrhea or amenorrhea (anorexia nervosa, excessive sport or ballet, etc.) is well known. However the prevalence of osteopenia among 16-18 years old girls with the cycle disorders mentioned above--caused by "pure" hypothalamo-pituitary-ovarian insufficiency--is not yet sufficiently examined. The hormonal (FSH, LH, prolactin, LH/FSH, estradiol. testosterone, progesterone) and ion (Ca++,PO4(3-),Na+,K+,Cl-) parameters and the bone mineral density (BMD) of the lumbar spine of 19 girls age 16-18 with oligomenorrhea or secondary amenorrhea, due to hypothalamo-pituitary-ovarian axis insufficiency were investigated, and correlation were searched for among them. In 3 of the case significant BMD reduction was found with a value lower than the -2 SD. compared to the age, sex and race matched control values, showing definite osteoporosis. The BMD of 10 girls was between the -2.SD. and -1 SD.: they had osteopenia. Only 6 of them had normal BMD ranging from the -1 SD. to the +1 SD. Neither the ion or hormonal values, nor the clinical parameters (height, weight age, age at menarche, duration of amenorrheic period) showed correlation with the BMD values, except of the body mass index (BMI), which showed a loose positive linear correlation. The measured low BMD values have a significance, referring to a possible reduction in the peak BMD. Patients having low peak BMD have an inclination for earlier, and more sever osteoporosis and fractures in the climacteric decades. These results emphasize the need of effective and early treatment of adolescent bleeding disorders from the point of view of prevention of osteoporosis as well. PMID- 9411344 TI - [Prevention of acute respiratory infections in healthy young adults by using oral immunomodulators]. AB - It is a general experience at army posts that right after the joining up period the number of airway infections suddenly increases. The upper and lower airway infections are especially facilitated by the confinement of military barracks, and the high number of smokers among soldiers. In the winter of 1995/96 at an army post of the Hungarian National Defense Forces authors treated 68 healthy, young recruits, aged between 18-23 preventively with an immunomodulant which contains lyophilized bacterial extracts, and placebo. The aim was to try this medicine on healthy adults to prevent or influence acute respiratory infections. In one third of the nine-month long follow-up period, a blind study was carried out, and two thirds of it were spent with a double blind one. The number of airway infections and off duty days was reduced by more than 40% in the group which was treated with the immunomodulant. In case of airway infection the complains and the signs were also milder in this group. Furthermore the medicine consumption decreased too. All of these indicate that the tried immunomodulant can be used in the primary prevention of airway infections among healthy adults even in a markedly infective environment. PMID- 9411345 TI - [Computerization and robotics in medical practice]. AB - The article gives the outlines of all principles used in computing included the non-electrical and analog computers and the artifical intelligence followed by citing examples as well. The principles and medical utilization of virtual reality are also mentioned. There are discussed: surgical planning, image guided surgery, robotic surgery, telepresence and telesurgery, and telemedicine implemented partially via Internet. PMID- 9411346 TI - [Incidence and mortality of aortic dissection]. AB - A population-based study was carried out over a 24-year period between January 1, 1973 and December 31, 1996 to determine the incidence and mortality of the aortic dissection in an administratively defined population of 106,000 inhabitants. The study comprises the consecutive cases of both inpatients and outpatients. Altogether, 74 cases of aortic dissection were found corresponding to 2.89%/year incidence. From the 74 patients 60 were admitted to hospital and 14 outpatients dead before the admission. In the first half of the observational interval (1973 84) 32 cases and in it's second half (1985-96) 42 cases were occurred with incidence of 2.50%/year and 3.28%/year, respectively. The higher incidence of the cases in the second half interval was explained by the significant increase of the autopsy rate of the dead outpatients. The male/female ratio was 1:1.6. The age ranged from 36 years to 97 years with a mean of 66.4 years and the women have had higher mean by the 7.0 years (p < 0.05). The mortality was explosively abrupt at the onset of the disease. Fourteen outpatients (18.9% of the total) died suddenly before admission and in the inpatients after the admission 28.4% died within the first hour, 44.6% within 12 hours, 62.2% within 24 hours and 75.5% within the first two days. Six patients were surgically treated and their perioperative mortality was 2/6, one month survived 4/6 and three years survived 3/6. The other 71 patients died. On the bases of the above-mentioned facts this work may be considered as an representative study. Hereby, the incidence of the aortic dissection in Hungary at least 3.0/100,000/year or rather 300 cases/year should be assessed. PMID- 9411347 TI - [Prevalence of Helicobacter pylori, correlations between alcohol consumption and gastroduodenal damage]. AB - We studied the prevalence of Helicobacter pylori (HP) infection, and that of upper gastro-intestinal tract lesions in chronic alcoholics. 73 chronic alcoholic, 74 duodenal ulcer and 40 non-ulcer dyspepsia patients were included. The prevalence and severity of HP infection and gastritis, profile of endoscopic lesions and the correlation between severity of alcohol consumption and that of HP infection were determined. HP was found in 31.5% of alcoholics, 83.78% of duodenal ulcer (p < 0.05) an 47.50% of non-ulcer dyspepsia cases (p > 0.18). The Genta score was in the same groups 0.47 +/- 0.09, 1.91 +/- 0.22 (p < 0.05), and 0.65 +/- 0.14 (p > 0.98). Endoscopy revealed oesophagitis in 23.6%, nonerosive gastritis in 24.5%, erosive gastritis in 13.6%, gastric ulcer in 17.5% and duodenal ulcer in 28.6% of alcoholics. Multiple findings were present in 51.8% and no lesion was found in 6.4% of the cases. Severity of gastritis was also lower in alcoholics as compared to duodenal ulcer patients. There was no correlation between severity of drinking and that of HP infection. Thus, heavy alcohol intake is associated with lower prevalence and severity of HP infection. However, alcoholics have significant, often multiple endoscopic lesions suggesting the role of other aetologic factors in addition to HP infection. PMID- 9411348 TI - [Study of the Leiden mutation (factor VQ 506), the most frequent cause of thrombophilia, in 116 thrombosis patients]. AB - The incidence of Leiden mutation was examined by PCR method in 116 thrombophilic patients in random fashion. Mean age at the first thrombotic episode was 30.97 years. 90 patients had positive family history for thrombosis, 67 had more than one thrombotic episodes. APC resistance with laboratory test was found in 51 cases (44%). F V Leiden mutation was proven in 44 patients (38%). 10 being homozygous and 34 heterozygous out of them. PMID- 9411349 TI - [Experience with ipriflavone therapy in postmenopausal osteoporosis]. AB - The data of 55 patients treated with ipriflavone are discussed. The protocol of examination and treatment are introduced. At the 12-month follow-up visit BMI increased with 3-5% in the lumbal spine and in the femur neck. Significant decrease of pain was observed in 55% of the patients. The number of side effects were low. According to the data, ipriflavone treatment was effective and well tolerated in postmenopausal osteoporosis. This treatment can be performed in cases when hormonal replacement therapy is contraindicated. PMID- 9411350 TI - [Microenvironmental lead poisoning]. AB - Authors describe two cases of lead poisoning of unusual origin. A 43 year old male patient bought some waste metal in order to build a fence around his new house for saving money. He removed the old corroded coat of paint using a grinder without any protection. Consequently, he had got grave lead intoxication with colic and anemia. Another 34 year old unemployed male person moulded leaded chessmen in his own kitchen. This activity resulted in serious intoxication of the patient and highly increased lead absorption of the family members. In addition to the exploration of professional exposure the authors stress also the significance of the clearing up of occasional lead-related hobbies and home activities in case of lead intoxication of unknown origin. PMID- 9411351 TI - [Forgotten years of the University of Kolozsvar]. PMID- 9411352 TI - [Remembering Janos Teofil Fabini on the 150th anniversary of his death]. PMID- 9411353 TI - [Recommendation on the uniform evaluation of the international and national scientific and medical literature]. PMID- 9411354 TI - [Interleukin-6 (IL-6)]. PMID- 9411355 TI - [The C-cell (parafollicular) of the thyroid: historical perspective]. PMID- 9411356 TI - [The workload in Pathological Anatomy measured on the basis of categories of services]. AB - Workload measurement is one of the most reliable tools in perspective payment systems, to evaluate clerk and technical personnel needs in a Surgical Pathology Department. In 1992 Italian Government began a full-comprehensive modification of financial support in National Health Care System, and during the last four years Public Hospitals and Local Health Units modified their financial organization with the introduction of workload measurement for the personnel needs. We propose a synthetic method for the workload measurement in Surgical Pathology Departments, similar to that proposed for perspective payment of surgical pathology services. The method is reliable and simple representing the full performance to obtain pathological report. PMID- 9411357 TI - [Malignant mesothelioma of the pleura. Morphometric study]. AB - Malignant mesothelioma is difficult to distinguish from other pleural malignancies and also from benign mesothelial lesions. A morphometric study has been performed to distinguish between them using quantitative size and shape parameters. Seven cases of malignant mesothelioma, 5 cases of pleural metastatic adenocarcinoma and 4 cases of benign mesothelial lesions were selected and subjected to S.A.M. (Shape Analytical Morphometry). The results, statistically evaluated, showed that morphometric parameters can be proposed for diagnostic purposes being useful in the discrimination among the three populations. In fact, multivariate discriminant analysis (MDA) of the quantitative parameters obtained by morphometrical study distinguished the three groups of lesions with only 2% of error between BML/MM, 7% of error between BML/MA and 25% between MM/MA. PMID- 9411360 TI - [Between the ideal and the real: autopsy in post-unification Italy]. AB - A short history of autopsy practice in Bologna since 1859 is presented. The programmes of the chiefs of the school of anatomic pathology are revisited. A summary of the scientific impact of autopsy practice is also illustrated. PMID- 9411358 TI - [DNA flow cytometry. Comparison of the methods for nuclear extraction from frozen and paraffin-embedded sections]. AB - Cellular DNA content of solid tumors can be determined either from fresh, frozen, or formalin fixed, paraffin-embedded tissues. However, discordant results have been obtained using the paraffin-embedded technique, and lack of abnormal DNA stemlines in the paraffin-embedded as compared to either fresh or frozen tissues has been reported. In this study we evaluated the validity of nuclear extraction method from paraffin-embedded tissues, using 75 breast carcinomas whose DNA content was previously analyzed from frozen tissue and resulted either normal (12 cases) or abnormal (63 cases). From representative paraffin blocks, nuclei were extracted following Hedley's technique. The results revealed excellent cell counting and good histogram resolution from all paraffin samples; the loss of G2M abnormal peak in eight histograms with abnormal stemline did not compromise the correct interpretation of DNA content. In addition, the comparison between DNA indices obtained from corresponding paraffin and frozen samples showed a good correlation in 69 cases (r = 94); discordance in six cases was demonstrated to be related to tumor heterogeneity. In conclusion the paraffin extraction method is a sensible, and reliable technique, which can be applied for DNA flow cytometric studies on archival cases, as well as whenever fresh sample from the tumor is not obtainable. PMID- 9411359 TI - [Comparative study of endocrine differentiation in early and advanced stomach carcinoma]. AB - To comparatively evaluate the frequency of the neuroendocrine differentiation in the early and in the advanced stage of the disease, 189 gastric carcinomas (67 early gastric cancers and 122 advanced gastric cancers) were studied by immuno histochemistry using a monoclonal antibody against chromogranin A (CgA). CgA positive tumor cells were detected in 55 of 189 gastric carcinomas (29.1%). Twenty-two of 67 early gastric cancer (EGC) (32.8%), and 33 of 122 advanced gastric cancer (AGC) (27.0%) were CgA positive. The latter included 23 intestinal type, 8 diffuse type and 2 mixed type tumors. The distribution of CgA-positive tumor cells in most AGC and EGC was focal and the endocrine cells were singularly scattered in the neoplastic glands or, more rarely, grouped in small clusters. CgA-positive tumor cells, moreover, were observed in the lymph node metastases of 12 AGC and 1 EGC. Statistical analysis of data showed no significant difference in the frequency of CgA-positive endocrine cells with regard to the stage of tumor growth, histotype, sex and age. These results indicate that neuroendocrine differentiation is a phenomenon independent of tumor stage, histotype, age, and sex and that CgA-positive cells are present during the whole neoplastic progression. PMID- 9411361 TI - [Primary malignant non-Hodgkin's lymphomas of the central nervous system in immunocompetent patients: diagnostic, prognostic and therapeutic criteria]. AB - Primary central nervous system non-Hodgkin's malignant lymphomas (PCNSNHML) have undergone a remarkable increase in incidence in the last years. Clinically PCNSNHML may present with the symptoms that mimic many others neurological conditions. Radiologically they appear as iso or hyperdense deposits, often multiple, usually supratentorial in periventricular region. Histologically although most lesions are high grade B cell lymphomas, T cell lymphomas are being recognised with increasing frequency. Actually a substantial improvement in survival has been obtained using chemotherapy as sole treatment, eventually followed by radiotherapy for recurrences. The prognosis, however, remains poor in comparison with nodal lymphomas. It is improved by T-cell phenotype, a Karnofsky performance status more than 50%, age at onset less than 50 years and negativity for bcl-2 oncogene of neoplastic cells. PMID- 9411362 TI - [Male diabetic mastopathy: description of a case]. AB - A case of diabetic mastopathy in a male is described. The patient is a 44 year old man with an insulin-dependent diabetes of long duration, presenting with a poorly defined nodule in his left breast. The lesion was constituted by a dense fibrous tissue with epithelioid stromal cells and a perivascular nodular lymphoid infiltrate. The differential diagnosis and the literature on diabetic mastopathy are briefly reviewed, particularly regarding male patients. The present case indicates that diabetic mastopathy may arise in the male breast. PMID- 9411365 TI - [Coronary embolism and myocardial infarction with heat rupture during infectious endocarditis]. AB - We report a case of transmural myocardial infarction due to coronary embolism complicating infective endocarditis in a 41 year old woman. The infarction, clinically silent, was followed by rupture of left ventricular free wall, leading to hemopericardium and cardiac tamponade. The case is an uncommon complication of infective endocarditis, at the same time an uncommon cause of myocardial infarction. PMID- 9411363 TI - [Subcutaneous myolipoma with bizarre cells: morphological, immunohistochemical and ultrastructural study of a case and review of the literature]. AB - A new case of subcutaneous myolipoma is reported. Grossly it appeared as a yellow whitish nodule of 5.5 cm, in its major axis and showed well defined borders. Histologically the tumour was constituted by two components: areas of mature fat tissue were interminged with more cellular areas composed of bundles of spindle shaped eosinophilic cells, reminiscent of smooth muscle cells. In these latter component, cells showing multilobated, bizarre nuclei were also focally evident. No areas of necrosis, nor mitosis were found in both the components of the lesion. The immunohistochemical and ultrastructural findings revealed the myofibroblastic nature of the cellular myoid component of the lesion. The presence of myoid cells with bizarre, multilobated nuclei was considered expression of a regressive phenomenon, as observed in the so called "bizarre" leiomyoma of the uterus. Two years after the initial diagnosis the patient is well and free of disease. Soft tissue myolipoma is a benign lesion which has to be distinguished from lesions with malignant or uncertain biologic behaviour. The clinicopathologic features of the present case are discussed and compared to those of the previously reported cases of soft tissue myolipoma. PMID- 9411364 TI - [Nevus of Ota. Presentation of a case associated with a cellular blue nevus with suspected malignant degeneration and review of the literature]. AB - BACKGROUND: Nevus of Ota is a melanotic pigmentary disorder ("dermal melanocytosis"), mostly congenital or acquired, involving the skin innervated by the first and the second branch of the trigeminal nerve, hence its descriptive label of "nevus fuscus coerulaeus ophthalmic and maxillaris". In more than half of patients this condition is associated with "ocular melanocytosis" ("melanosis oculi") involving the conjunctiva, the sclera, the uveal tract and possibly the optic nerve. In some cases a condition of "orbital melanocytosis" with involvement of orbital fat and periosteum by dendritic melanocytes is on record as well as in some other patients an analogous condition of "leptomeningeal melanocytosis" is present. At histology tissues from the above sites are seen infiltrated by dendritic melanocytes which can vary in number from so scarce up to so numerous that a diagnosis of a blue nevus of the common type is warranted. Sometimes the finding of a variously pigmented typical cellular blue nevus in the skin and alternatively that of heavily pigmented melanocytoma in the eye ("nevus magnocellularis") or in the meninges (so-called "melanotic meningioma") are respectively observed. MATERIALS AND METHODS: A case of cellular blue nevus with histologically uncertain malignant potential in a nevus of Ota of 30 years duration in a white female patient aged 59 is described. The lesion which was surgically totally removed grossly appeared nodular shaped and 2 cm sized. Histologically it consisted of a fairly well-circumscribed proliferation of melanocytic spindle-shaped cells growing in a vaguely fascicular pattern. On the basis of random nuclear atypicalities and pleomorphism and additionally by virtue of the presence of a few scattered mitoses (one of which was atypical) but in absence of frank necrosis a diagnosis of unpredictable biologic behaviour seemed to be warranted. The patient was closely followed-up but no adjunctive therapy given. Four years after the excision and diagnosis no local recurrence or distant metastasis has been discovered. A computerized search of previously recorded cases of melanomas in nevus of Ota was made. CONCLUSIONS: Forty-eight cases of malignant melanomas complicating this clinico-pathological setting are on record, mostly in the uveal tract, followed by locations in central nervous system, skin, and retro-orbital fatty tissue. Melanomas arising in nevus of Ota tend to be low grade lesions that do infiltrate locally but rarely metastasize. The importance of a closely dermatological and ophthalmic surveillance of patients with nevus of Ota is emphasized. PMID- 9411366 TI - [Total anomalous pulmonary venous drainage. Description of a case]. AB - A case of total anomalous pulmonary venous drainage (TAPVD) is reported, which according to Darling classification corresponded to infradiaphragmatic type (type III) and is associated to pulmonary arterial thrombosis and infarctions. It's emphasized the need of an accurate dissection of cardiovascular system in perinatal necropsies, specially in cases of sudden infant death. By an appropriate dissection, cardiovascular malformations can be recognized. Moreover, it's explained the pathogenesis of pulmonary arterial thrombosis and infarctions. PMID- 9411368 TI - [An unusual case of intramuscular hemangioma]. AB - A case of cavernous-capillary intramuscular haemangioma is reported. The tumour was characterized histologically by a proliferation of capillaries, cavernous blood spaces, along with arterial and venous blood vessels, intermingled with striated muscle fibres and fat tissue. The complex mixture of vessels and the circumscribed margins of the tumour could support a congenital origin. PMID- 9411367 TI - [Malignant paraganglioma of the posterior mediastinum with 27 years follow-up]. AB - INTRODUCTION: A costovertebral paraganglioma with a long history of recurrences and metastases is reported. MATERIAL AND METHODS: The lesion is documented on histological ground and with immunocytochemistry and electron microscopy. RESULTS: The patient is a 42 year old man which presented with a 5 cm axillary mass and enlarged mediastinal lymph nodes. The former lesion has been interpreted as a metastasis of paraganglioma. The patient underwent surgery because of a costovertebral paraganglioma at the age of 19 and showed locoregional metastases and a recurrence over a period of 23 years. Twenty-seven years from removal of the primitive tumor, the patient is alive and well. All the neoplasms showed identical histological features as they were composed of neoplastic elements arranged in nests surrounded by dendritic cells. Neoplastic cells immunoreacted with anti-chromogranin antiserum whereas sustentacolar cells were positive with S 100 antiserum. DISCUSSION: The differential diagnosis between a multicentric paraganglioma a metastasis has been particularly taken into consideration. In addition, axillary tumor was distinguished from a metastatic renal cell carcinoma and melanoma. The literature regarding costovertebral paraganglioma has been also revised. PMID- 9411369 TI - [Who needs a gastroenterology pathologist?]. PMID- 9411370 TI - [Who needs a gastroenterology pathologist?]. PMID- 9411371 TI - [Who needs a gastroenterology pathologist?]. PMID- 9411372 TI - [Who needs a gastroenterology pathologist?]. PMID- 9411373 TI - [Intra-epithelial melanocytic neoplasia (MIN), a new term proposed for dysplastic melanocytic lesions]. PMID- 9411374 TI - [HIV-1 virus uses a double-key mechanism to infect the cell]. PMID- 9411375 TI - [Diagnostic methods in nephropathology: the necessary and the superfluous]. PMID- 9411376 TI - [www.siapec.it...which one reads: World Wide Web, SIAPEC, Italy...]. PMID- 9411377 TI - [Use of the Internet by anatomo-pathology specialists]. PMID- 9411378 TI - [Luigi Sacco and the history of smallpox in Italy]. PMID- 9411379 TI - [The role of the anatomist/pathologist in modern medicine]. PMID- 9411380 TI - United States vaccine research: a delicate fabric of public and private collaboration. National Vaccine Advisory Committee. AB - In the last 20 years, two thirds of all new vaccines provided worldwide have been produced by a US network of independent industrial, governmental, and academic partners engaged in vaccine research and development. Vaccines are complex products and the science of vaccinology is difficult. To achieve the full promise of modern science and technology to prevent and treat disease by immunization, the delicate fabric of America's cooperative and collaborative vaccine research relationships must be sustained and strengthened. The major partners are the federal government; four large companies--two US-headquartered (Wyeth-Lederle Biologics and Vaccines and Merck & Co), two foreign firms (SmithKline Beecham and Pasteur Merieux Connaught); and academia. Of the $1.4 billion that fund US vaccine research and development annually, 46% comes from vaccine sales, 36% from taxpayers, and 18% from risk capital. Vaccine innovation could be strengthened by improved public and policy maker understanding of the vaccine development network; declarations of partnership; interactive dialog with federal advisory bodies; public forums for government and industry to listen to patients, providers and researchers; sabbatical assignments between partners; mechanisms to share industries' market research with public immunization programs; continued active industry participation in the Advisory committee on Immunization Practices and the National Vaccine Advisory committee; increased collaboration between industry and the National Institutes of Health for clinical research; harmonization of the Advisory Committee on Immunization Practices vaccine recommendations and the Food and Drug Administration package inserts; and public policies to foster the partnership's collaboration and robustness. The optimal size and configuration of the US vaccine enterprise should be debated only in the context of a full understanding of how the current system works and its record of effectiveness. These National Vaccine Advisory Committee recommendations are directed at developing public policies to foster and sustain vaccine innovation and ensure the timely introduction and supply of new vaccines needed by this nation and the world. PMID- 9411381 TI - Breastfeeding and the use of human milk. American Academy of Pediatrics. Work Group on Breastfeeding. AB - This policy statement on breastfeeding replaces the previous policy statement of the American Academy of Pediatrics, reflecting the considerable advances that have occurred in recent years in the scientific knowledge of the benefits of breastfeeding, in the mechanisms underlying these benefits, and in the practice of breastfeeding. This document summarizes the benefits of breastfeeding to the infant, the mother, and the nation, and sets forth principles to guide the pediatrician and other health care providers in the initiation and maintenance of breastfeeding. The policy statement also delineates the various ways in which pediatricians can promote, protect, and support breastfeeding, not only in their individual practices but also in the hospital, medical school, community, and nation. PMID- 9411382 TI - Scope of health care benefits for newborns, infants, children, adolescents, and young adults through age 21 years. American Academy of Pediatrics. Committee on Child Health Financing. AB - The optimal health of children can best be achieved by providing access to comprehensive health care benefits. This policy statement replaces the 1993 statement, "Scope of Health Care Benefits for Infants, Children, and Adolescents Through Age 21 Years." Changes involve services and procedures specific to the delivery of comprehensive preventive, prenatal, postnatal, and mental health care. These services should be delivered by appropriately trained and board eligible/certified pediatric providers, including primary care pediatricians, pediatric medical subspecialists, and pediatric surgical specialists. PMID- 9411383 TI - Revised group B streptococcal (GBS) infection guidelines. PMID- 9411384 TI - Thyroid function 6 years after prenatal treatment with thyrotropin-releasing hormone. PMID- 9411385 TI - Respiratory syncytial virus PREVENT Study questions. PMID- 9411386 TI - Respiratory syncytial virus PREVENT Study questions. PMID- 9411387 TI - Respiratory syncytial virus PREVENT Study questions. PMID- 9411388 TI - Sedation for procedures. PMID- 9411389 TI - Sedation for procedures. PMID- 9411390 TI - Sedation for procedures. PMID- 9411391 TI - Childhood lead poisoning from apple cider. PMID- 9411392 TI - Role of calmodulin as a regulator of Ca2+-dependent exocytosis in adrenal chromaffin cells. PMID- 9411393 TI - [Delaying aging--a possibility of wishful thinking]. PMID- 9411394 TI - [Apoptosis: molecular aspects]. AB - Many signals and external stimuli regulate the apoptosis activity by interaction with the genome. These stimuli include morphogenetic signals, physiological factors, and environmental influence. The signals mediate their effect on cells with suitable receptors, relevant signalling pathways, and competence to execute the apoptosis cascade. Apoptosis is triggered indirectly by deprivation of survival factors, or directly by intercellular cell death signalling factors, and also by unbalanced intracellular messenger molecules, which are, more or less, involved in regulation of both programmed cell death and survival. Several genes are involved in regulation of cell survival and apoptosis: bcl-2/bax, p53, c-myc and transcription factors such as cdk, c-myc, c-fos and c-jun. Apparently, apoptosis could be triggered by increased or inhibited gene expression as well as biochemical reactions without changed gene expression. The morphological changes during apoptosis reflect a cascade of genetic and biochemical reactions in the cell. In the signal transduction pathway both secondary messenger Ca2+, different kinases, and polyamines are involved. Cysteine proteases cleave cytoskeletal proteins, endonucleases divide DNA into fragments, and transglutaminases cross link macromolecules. Degradative enzymes such as proteases, endonucleases and transglutaminases are activated during apoptosis, leading to cellular collapse and formation of vesicular apoptotic bodies. Both increased and inhibited apoptosis activity may have pathological consequences. New therapeutic strategies aim to counteract dysregulation of apoptosis in specific tissues by pharmacological intervention. Thus there is a need for identification of molecules and gene products involved in regulation of apoptosis activity and clarification of the conditions where this knowledge may be used. PMID- 9411395 TI - [Postmenopausal hormone substitution--for many of for few? I. Effects of long term hormone substitution]. AB - Users of hormone replacement therapy (HRT) are characterised by a 20-50 per cent reduction in all-cause mortality, and a 50 per cent reduction in the risk of coronary disease and osteoporotic fracture. The risk of Alzheimer's dementia or of cerebrovascular disease has also been reported to be reduced, though published findings have been inconsistent. In contrast, the risk of breast cancer or venous thrombosis is probably increased. A number of minor side-effects are associated with HRT, though hitherto their impact has received little attention. Although randomised controlled studies have been conducted to assess relationship between HRT and risk factors for disease, no such studies of relationship between long term HRT and myocardial infarction, osteoporotic fractures or mortality have been performed in large series. Selection bias may thus explain part of the reported beneficial effects of HRT on morbidity and mortality found in observational studies. PMID- 9411396 TI - [Laparoscopic surgery--where do we stand?]. AB - Owing to the rapid advances in laparoscopic surgery during the past decade, almost all abdominal surgery has now been done with laparoscopic or laparoscopy assisted procedures. However, there is still no international consensus as to the indications for the general use of laparoscopic surgery, as compared with open surgery, and the hitherto insufficient analysis of the economic consequences of transition to laparoscopic surgery. There is also a lack of good comparative studies of open surgery and laparoscopic surgery performed in the context of optimised multimodal perioperative treatment with adequate pain relief, early oral nutrition and mobilisation, and recommendations for shortened absence from work. PMID- 9411397 TI - [TOS (thoracic outlet syndrome)--a challenge to conservative treatment]. AB - Functional impairment and pain in the upper extremities may indicate a functional deficit in the thoracic outlet. Static work posture, trauma and whiplash injury may be predisposing factors. The younger generation who often spend long hours in front of a computer are in danger of becoming a future risk group. The primary care physician should be familiar with the syndrome which can be identified by careful clinical examination. Timely intervention can prevent much of the disabling symptomatology. Treatment is primarily conservative and should be aimed at the restoration of functional capacity. As in other disorders, the individual constellation of symptoms is dependent on circumstantial factors, an aspect meriting particular attention in treatment and follow-up. Optimisation of ergonomic conditions is important feature of treatment, and long-term follow-up is necessary. Transient exacerbation is not an indication for surgical treatment. If cervical and thoracic outlet function has normalised but the patient still has symptoms, then the differential diagnosis should be reconsidered. Examination and treatment of patients with pain in the upper extremities requires the collaboration of the physician and physical and occupational therapists. Treatment can be delivered in the primary care setting. PMID- 9411398 TI - [From risk factors to health resources. From theory to medical practice]. AB - In this article, we emphasize some of the problems related to the choice of risk factor identification and intervention as the dominant medical strategy for diagnosis, treatment and prevention of disease. Knowledge about risk factors can provide hypotheses about origins of disease and predict disease at a group level. However, there are several pitfalls related to this perspective concerning causal factors, end points, informed consent, group level based possibilities and medicalization. A salutogenic perspective shifts the attention toward resources which might counteract risk of disease. Increased attention to people's personal health resources--assessed by the doctor as well as by the person herself--can contribute to a better balance in medical theory and practice by stimulating the strength of people rather than looking only for weaknesses. PMID- 9411399 TI - [Visiting special dwellings in Denmark]. PMID- 9411400 TI - [Admission routines in Norwegian nursing homes]. PMID- 9411401 TI - [Principles for longterm care in Finland]. PMID- 9411402 TI - [Nursing institutions fo the aged in Island]. PMID- 9411403 TI - [To be admitted to nursing homes in Sweden]. PMID- 9411405 TI - Arrhythmia risk stratification. Proceedings of the IVth Interlaken Symposium. February 1997. PMID- 9411404 TI - Grades of dispute. PMID- 9411408 TI - [Dobutamine stress echocardiography in the monitoring of post-angioplasty myocardial contraction and in detection of restenosis]. AB - The expanding role of percutaneous transluminal coronary angioplasty (PTCA) in the selected patients with coronary artery disease (CAD), requires the noninvasive method for the assessment of the influence of PTCA on left ventricular function and the detection of restenosis. Forty two patients (pts) with angina, undergoing PTCA, were evaluated with dobutamine stress echocardiography (DSE) 1 day before, 2-7 days after, 3-4 months and 1 year after successful angioplasty. Two-dimensional echocardiographic images were collected during incremental doses of intravenous dobutamine infusion (up to 40 micrograms/kg/min). In 31.7% of examinations atropine was added (0.5-1 mg). Echocardiograms were assessed at baseline and during low- and high-dose of dobutamine infusion. An echocardiographic test positive for restenosis was defined as showing a new wall motion abnormality induced by dobutamine 3-4 months or 1 year after PTCA, compared to DSE 2-7 days after PTCA. Whereas before angioplasty, the wall motion score index (WMSI) increased from 1.40 +/- 0.34 at rest to 1.56 +/- 0.34 at peak stress (p < 0.0001); after angioplasty WMSI decreased from 1.29 +/- 0.30 at rest to 1.24 +/- 0.28 at peak (p < 0.0001). In 20 patients control angiography (CA) was performed because of positive DSE (11/20) or for clinical reasons only (9/20). In this selected group CA showed significant restenosis in 10 patients with high DSE sensitivity and specificity, 83% and 91% respectively. In this group peak stress WMSI increased from 1.21 just after PTCA to 1.60 during the examination detecting restenosis (p < 0.01). CONCLUSION: 1. A reduction in myocardial ischaemia, as assessed by DSE, is seen early after angioplasty. 2. Serial DSE records changes in regional function, thus being a valuable tool in PTCA follow up. PMID- 9411406 TI - Pediatric emergency medicine: legal briefs. PMID- 9411409 TI - [Right ventricular diastolic disfunction and its relation to left ventricular performance in patients with hypertension]. AB - Abnormalities in left ventricular (LV) diastolic function may be the earliest indications of hypertensive heart disease. Because the two ventricles influence each other's performance this study was designed to investigate the impact of chronic LV pressure overload in essential hypertension (HT) on diastolic function of right ventricle (RV). RV and LV diastolic function was evaluated in 74 patients with mild-to-moderate essential HT using pulsed wave Doppler echocardiography. Fifty-five normotensive patients without heart disease acted as control subjects. In studied group, 17 patients (23%) had normal mitral (MV) and tricuspid (TV) flow parameters, 28 (38%) had impaired LV filling parameters [MV early (E) to late (A) peak flow velocity ratio (MV E/A) 0.81 +/- 0.12 vs control 1.19 +/- 0.18, p < 0.001] while 29 patients (39%) had abnormal both mitral [MV E/A) 0.72 +/- 0.15 vs control 1.19 +/- 0.18, p < 0.001] and tricuspid flow parameters (TV E/A) 0.8 +/- 0.19 vs control 1.23 +/- 0.1, p < 0.001). In group with impaired diastolic filling of both ventricles indices of mitral flow were significantly more abnormal compared to group with normal TV flow parameters (MV E/A 0.72 +/- 0.15 vs control 0.81 +/- 0.12, p < 0.05). RV filling parameters correlated with filling parameters. There was good correlation between TV A and MV E (r = -0.56, p < 0.01), the time velocity integral of early mitral inflow (MV E-VTI) (r = -0.64, p < 0.001) and positive correlation with MV A (r = 0.78, p < 0.0001). Also there was good correlation between LV mass and TV E (r = -0.56, p < 0.01) and the time velocity integral of early tricuspid inflow (r = -0.72, p < 0.001). Data indicate that RV diastolic function is abnormal in essential hypertension and these abnormalities are closely related to those of LV diastolic function and LV mass. PMID- 9411407 TI - [The influence of thrombopoietin (TpO) on human erythropoiesis. In vitro studies- clinical implications]. AB - The role of thrombopoietin (TpO) in regulations erythropoiesis is still controversial. This fact prompt us to present our own data. We found, that TpO is not able to replace erythropoietin (EpO) in stimulation of the growth of human erythroid colonies. TpO was found to be only a very weak costimulator of EpO dependent erythroid progenitors growth. This effect is however much weaker than those observed after costimulation with kit ligand or interleukin-3. The rationale for TpO application for treatment of anemia in humans seems therefore doubtful. This conclusion seems now to be supported by the results of the first trials performed in anemic patients. PMID- 9411410 TI - [The effect of amlodipine on structure and function of the heart and exercise tolerance in patients with hypertension]. AB - The aim of the study was to assess whether the hypotensive activity of amlodipine is associated with regression of left ventricular hypertrophy and improvement of impaired LV or right ventricular (RV) diastolic function or increasing of tolerance of physical activity in hypertensive patients. Assessment of left ventricular structure, systolic and diastolic function as well as RV diastolic dimension and diastolic function were performed in 24 patients with mild-to moderate hypertension before administration of amlodipine and 3, 6 and 9 months of the treatment. In order to assess the tolerance of physical activity, incremental treadmill exercise testing was performed at baseline and after 6 and 9 months of the therapy with amlodipine. RESULTS: During 9 months of the therapy with amlodipine no significant change in indexes of LV mass or in LVM was observed. Similarly amlodipine did not influence the parameters of LV or RV diastolic function in studied patients. However, amlodipine treatment resulted in significant increase in total exercise time (p < 0.05), total workload (p < 0.01) measured in METs and decrease in diastolic blood pressure during exercise test. CONCLUSION: The nine months of the therapy with amlodipine resulted in significant improvement in exercise tolerance. Total exercise duration and total workload measured in METs significantly increased. During this time of the therapy no significant changes in LV structure or LV and RV diastolic function were observed. One can make an assumption that amlodipine inhibits progression of structural and functional derangement in hypertensive patients. PMID- 9411412 TI - [Echocardiographic evaluation of cardiac structures in patients with rheumatoid arthritis]. AB - Since heart lesions were found at autopsy in 55-60% of rheumatoid arthritis patients, we decided to assess echocardiographically their clinical significance. The study comprised 100 consecutive patients with rheumatoid arthritis (77 females and 23 males) of the mean age 55.7 +/- 12.5 yrs (range 18-83 yrs) and the disease duration of 8.3 +/- 8.0 yrs (range 1-35 yrs). The control group consisted of 100 consecutive age and sex matched patients admitted to university hospital. All the patients underwent echocardiographic examinations in apical and parasternal projections. The activity of the rheumatoid process, the severity of articular lesions, the presence of extraarticular sings as well as HLA DR and DQ antigens were determined clinically and with laboratory tests. Twenty six patients with rheumatoid arthritis had pericardial effusion, 10 revealed the sings of chronic pericarditis, in the control group 4 and 0 respectively (p = 0.001 and p = 0.025). No difference was shown in the wall contractions disturbances, size of the cardiac cavity, or thicknesses of the interventricular septum or posterior wall. In 3 rheumatoid arthritis patients, a valvular heart disease was diagnosed, this number was not significantly different from that in the control group (2 patients). There was no correlation between the lesions observed in the heart and the rheumatoid process activity estimated with clinical and laboratory indices. PMID- 9411411 TI - [Usefulness of CA 15-3 antigen determination for evaluation of response to second line chemotherapy in patients with breast cancer: preliminary study]. AB - The aim of this study was to assess effects of second-line chemotherapy in metastatic breast cancer via determination of CA 15-3 marker. Analysis included 73 women, in whom distant metastases were diagnosed within 14-72 months (median: 43) after the completion of basic therapy. Average age of patients at primary diagnosis was 50.7 +/- 12.6 years. Dominant sites of metastases were: liver (27 patients) and lungs (24 patients). Serum CA 15-3 was examined immunoenzymatically at diagnosis of distant metastases and then after 2-4 cycles (median 4) of anthracycline-based chemotherapy. Changes of mean CA 15-3 values correlated with the UICC response criteria. There was a significant fall in mean levels of CA 15 3 after treatment in patients with complete (p < 0.004) and partial regression of metastatic lesions (p < 0.03). Stabilization and progression of the disease were associated with raise in CA 15-3 mean values, but the difference was significant only in the latter group (p < 0.02). In 31 out of 39 patients (79.5%) with regressive disease (complete and partial response) CA 15-3 levels decreased by at least 25% after treatment. Nine of 11 (81.8%) patients with stable disease had the antigen concentrations that did not vary by more than +/- 25% of the initial CA 15-3 value. CA 15-3 levels raised by at least 25% in 22 out of 23 (95.7%) cases with progressive breast cancer. Overall, CA 15-3 variations correlated with the disease status in 62 (84.9%) patients. These findings confirmed the usefulness of CA 15-3 determinations in evaluating the efficacy of second-line chemotherapy in patients with advanced breast cancer. PMID- 9411413 TI - [Subcutaneous immunoglobulin infusion in antibody deficiency substitution of a patient sensitized to intravenous immunoglobulins. Case report]. AB - Patients with severe form of common variable immunodeficiency require chronic immunoglobulin substitution. However, intravenous Ig administration may not be possible in some of them because of serious anaphylactoid reactions. It has been suggested that such patients may tolerate well Ig administration by subcutaneous infusion. A case is described (originally with IgG level of 53 mg/dl) who reacted with anaphylactic shock to intravenous immunoglobulin and now for more than 7 months at 1-2 week intervals receives immunoglobulin by subcutaneous infusion without any adverse reactions and maintaining IgG level above 400 mg/dl. In contrast to the period proceeding immunoglobulin substitution, the patient remains free of bacterial infection during last 7 months. PMID- 9411414 TI - [Some factors affecting longevity and quality of life in patients treated with continuous ambulatory peritoneal dialysis (CAPD)]. PMID- 9411415 TI - [Hypercoagulable state in diabetes mellitus]. PMID- 9411416 TI - [New therapeutic perspectives associated with the introduction of angiotensin II receptor antagonists]. PMID- 9411417 TI - [Forgotten therapeutic methods in material from the Polish Archives of International Medicine from the years 1923-1939]. PMID- 9411418 TI - [Peptide C and insulin after delivery in women with gestational diabetes]. AB - Gestational diabetes is a disease appearing in many forms. Up till now the etiopathogenesis was not clearly defined. It has been suggested that counterregulatory of pregnancy or diminished B-cells could the major contributory factors. The aim of the present study was to retrospective verify the diagnoses of gestational diabetes. The investigation was carried out in 42 women aged 25-39 yrs, mean age 30 +/- 6 yrs. Diabetes was diagnosed in the 2nd, and 3rd, trimesters of pregnancy, 30 women were treated by diet only, 25 kcal/kg b.w. depending on weight, and 12 patients had intensified insulin therapy of mean daily dose 18 +/- 8 U. Three to nine months after delivery a glucose tolerance test as well as estimation of C-peptide and insulin concentration by RIA in basic conditions and after administration of 1 mg of glucagon were performed. In the group of women treated by diet only, normal values of glycaemia in glucose tolerance test were observed. C-peptide concentration measured before administration of glucagon was 1.15 +/- 0.49 ng/ml and after administration of 1 mg of glucagon was 3.14 +/- 1.44 ng/ml. In the majority of patients treated during pregnancy with insulin the results of oral glucose tolerance test were pathological. The concentrations of C-peptide in the test with glucagon were significantly lower (0.33 +/- 0.16 and 0.38 +/- 0.32 ng/ml). The concentrations of insulin were much lower in comparison to women treated with diet and healthy controls, these results suggest that, if gestational diabetes could be controlled by diet only, disturbances of carbohydrate metabolism would disappear, however, if insulin therapy was necessary during pregnancy, disturbances of the carbohydrate metabolism would be prolonged. PMID- 9411419 TI - [Level of neopterin in blood serum in selected thyroid diseases]. AB - The aim of our study was to estimate the levels of neopterin (NTP), anti thyroglobulin antibodies (ATG), anti thyroid microsomal antibodies (MAB), immunoglobulins (IgG, IgA and IgM) and creatinine in the serum of patients with thyroid diseases. The clinical material consisted of 71 patients (64 women and 7 men) with non-toxic nodular goitre (30 persons), neoplasms (33 persons: 22 with thyroid adenoma and 11 with thyroid carcinoma) and thyroiditis (8 persons). The control group consisted of 30 healthy women. In studied groups the mean serum levels of NTP and creatinine were in normal range. The highest levels of NTP were found in patients with thyroid carcinoma (7.14 +/- 5.95 nmol/l) and with non toxic nodular goitre (5.13 +/- 3.57 nmol/l). It was statistically significant in comparison with control group (p < 0.05). Anti thyroglobulin antibodies (ATG) were detected in serum of 36 patients (50.7%) and anti thyroid microsomal antibodies (MAB) were in serum of 28 patients (39.4%). Mean levels of these autoantibodies were significantly higher than in control group (< 0.05). In patients with autoantibodies (ATG and/or MAB) mean serum level of NPT was significantly higher than in patients without these autoantibodies (p < 0.05). We did not find a significant correlation between the level of NPT and the levels of ATG and MAB. PMID- 9411420 TI - [Monitoring of changes in levels of endogenous TNF alpha in patients with uncontrolled insulin dependent diabetes mellitus]. AB - In 27 patients with uncontrolled type 1 diabetes the changes of the endogenous TNF alpha serum concentration in correlation with blood glucose level were investigated. In the first 24 hours period, the concentration of glucose was analysed in the following way: immediately after the admission, next every hour for three hours and then after 6, 9, 12, 18 and 24 hours. In the followed days the blood glucose was determined five times a day. The serum concentration of endogenous TNF alpha was monitored immediately after the admission, after first hour of treatment and next after 6, 24, 72 hours. The control group consisted of 23 healthy persons. Glycaemia level and TNF alpha concentration were analyzed with Student-1 test for combined variables and the correlation Spearmann test. The level of TNF alpha markedly decreases together with the decrease of blood glucose level in each patient. The level of TNF correlates with the level of glycaemia (linear correlation). The results presented above unequivocally show that there is a correlation between endogenous TNF alpha and glucose level in serum of the patients with type 1 diabetes (at the decompensation stage of the disease). PMID- 9411421 TI - [Topographic differentiation of transperitoneal glucose transport: in vitro studies]. AB - The dynamics of bidirectional glucose transport across parietal peritoneum from anterior abdominal wall or diaphragm and small-bowel mesentery was analyzed. The mean transport values, expressed as transport coefficients, of intact peritoneum amounted 2.61; 0.18; 5.43 (10(-4) x cm x S-1), respectively. The chemical destruction of the mesothelium (2.5 mmol/l sodium deoxycholate) increased transperitoneal glucose transport across the investigated tissues. However, the intensification of transfer was different. Removal of the diaphragm pleural mesothelium caused greater increase of bidirectional glucose transport than the peritoneal mesothelium. Furthermore, the mean values of transport coefficient after peritoneal mesothelium damage were higher in relation to mesentery than to the peritoneum isolated from anterior abdominal wall and diaphragm. The obtained results suggest topographic heterogeneity of peritoneal membrane which refer to the mesothelium and submesothelial layers. PMID- 9411423 TI - [Morbidity of multiple myeloma in material from the Hematology Clinic AM in Lublin after the accident in Chernobyl]. AB - The average dosage of radiation which was measured in Poland during the year after damage of the nuclear power in Chernobyl (according to UNSCEAR) was 0.27 mSv, which gives 11% natural radiation dosage in the period of one year (2.6 mSv). Disturbances of cells genome caused by radiation are possible because of big dosage of radiation in Lublin region. It was interesting to define morbidity and mortality of multiple myeloma (MM) after the damage in Chernobyl. The average latent period of MM is about 20 years (like thyroid carcinoma). The increase of thyroid carcinoma morbidity after the damage of Chernobyl nuclear power plant in Byelorussia. Ukraine, Russia was observed. The increased morbidity rate of MM among patients of Haematologic Department (especially in the third stage of disease) and the increased mortality rate in Lublin region was confirmed. PMID- 9411422 TI - [Risk factors for coronary heart disease in 1002 patients with hyperlipidemia]. AB - The aim of this study was to estimate the coexistence of risk factors for coronary heart disease (CHD) in hyperlipidemic patients. Studies were performed in 1002 (601 women, 401 men) subjects who referred to our outpatient clinic among 12 months. Hypercholesterolemia was the predominant lipid disorder found in 66% of patients, mixed hyperlipidemia in 31.8%, and hypertriglyceridemia only in 2.2%. Overweight and obesity remain a major health burden among our patients: BMI > or = 25 was observed in 66%. Hypertension was recognized in 37.5% of subjects, and diabetes mellitus in 11.2%, 17% were long-term smokers. Familial aggregation of hyperlipidemia was observed in 15.7% of subjects, and more than 44% had a positive family history of cardiovascular disease. Low HDL cholesterol levels (< 35 mg/dl) were seen frequently in men (24.7%) and rare in women (7%). Lp(a) excess (> or = 30 mg/dl) was observed in 12% of patients. Myocardial infarction (MI) had already 11.7% subjects (7% women, 18.7% men). In these patients CHD risk factors were observed more frequently. The higher apo B and Lp(a) levels and lower HDL cholesterol levels were recognized in the patients who suffered from MI. More than 83% of our hyperlipidemic patients had coexistence CHD risk factors. The multiple coexisting risk factors cause the high risk for CHD and they require intensive correction. PMID- 9411424 TI - [Marked polyglobulia in patients with obstructive sleep apnoea]. AB - Two patients with obstructive sleep apnoea are presented. Both had marked hypoxaemic polycythaemia treated for many years with phlebotomies before a proper diagnosis was established. Diagnosis of obstructive sleep apnoea and introduction of CPAP treatment resulted in clinical improvement and regression of polycythaemia. In secondary polycythaemia differential diagnosis should include obstructive sleep apnoea. PMID- 9411425 TI - [Pulmonary reaction after furazidin (Furagin). Case report]. AB - For the first time in Poland we present the case of pulmonary reaction to furazidin which is by chemical structure closely related to nitrofurantoin. 63 years old woman presented generalized symptoms of acute hypersensitivity reaction induced by furazidin as well as features of chronic pulmonary fibrosis. After few months of treatment with this drug patients complained of weight loss, dyspnea on effort, non-productive cough, chills and fever. Radiological and functional evaluation of respiratory system confirmed features of lung fibrosis. Drug provocation test was positive. In vitro furazidin in low concentrations stimulated proliferation of patient's lymphocytes. After cessation of treatment we have observed rapid improvement of clinical, radiological, biochemical and functional parameters. PMID- 9411426 TI - [Molecular aspects of the seroconversion from HBeAg -->anti-Hbe in the course of type B viral hepatitis caused by mutant strains the virus]. PMID- 9411427 TI - [Viral hepatitis B in the presence of mutant HBV strains of the "e-minus" type- clinical aspects]. PMID- 9411429 TI - [Professor Gerard Jonderko--life and achievements of the honorary member of the Polish Society of Internal Medicine]. PMID- 9411428 TI - [Immunology of ulcerative colitis--interactions of T lymphocytes with extracellular matrix proteins]. PMID- 9411430 TI - [Polish Society of Internal Medicine]. PMID- 9411431 TI - ["X"--the last unknown of hepatitis B? Transactivation and oncogenic properties of HBV X protein]. PMID- 9411432 TI - [Intracellular ATP-binding proteins]. PMID- 9411433 TI - [Changes in the regulatory domain of the myosin head observed in some heart diseases]. PMID- 9411434 TI - [Better late than never: discovery of MHC restriction awarded with the Nobel Prize after 22 years (Nobel Prize for physiology or medicine in 1996)]. PMID- 9411435 TI - [Synergistic effects of immunomodulators of the muramyl peptide type and other drugs used together with chemotherapy]. PMID- 9411436 TI - [Episialin--a newly recognized component of glycocalyx and cell membrane in epithelial cells]. PMID- 9411437 TI - [The influence of DNA damage on cell cycle regulation]. PMID- 9411438 TI - [The ubiquitin conjugation system and its role in the Saccharomyces cerevisiae yeasts]. PMID- 9411439 TI - [Coordination of leading and lagging DNA in strand replication]. PMID- 9411440 TI - The certainty of uncertainty: in spite of our best efforts, we can't always explain the outcomes. PMID- 9411441 TI - The effect of prolonged imipramine treatment on the alpha 1-adrenoceptor-induced translocation of protein kinase C in the central nervous system in rats. AB - The aim of the study was to investigate the effects of prolonged treatment with imipramine (10 mg kg-1/day i.p. for 21 days) on the translocation of protein kinase C (PKC) after stimulation of the alpha 1-adrenoceptor. Methoxamine (5-50 mg kg-1) and phenylephrine (0.1-1 mg kg-1) induced a rapid and long-term redistribution of PKC from the cytosolic to the membrane fraction in the rat frontal cortex and hippocampus. The effects of methoxamine and phenylephrine were completely blocked by pretreatment with prazosin. Prolonged pretreatment with imipramine changed the response of PKC to methoxamine and phenylephrine. Much lower doses of alpha 1-adrenoceptor agonists were able to induce the redistribution of PKC. Moreover, prolonged treatment with imipramine markedly increased the basal activity of PKC in the membrane fractions of the frontal cortex and hippocampus. PMID- 9411442 TI - [Anti-leukotrienes: a new perspective in treatment of bronchial asthma]. PMID- 9411443 TI - [alpha 1-Pi deficiency. World Health Organization]. PMID- 9411445 TI - [Gold-induced pulmonary disease: clinical features, outcomes, and differentiation from rheumatoid lung disease]. PMID- 9411444 TI - [Inhaled salmeterol or oral theophylline in nocturnal asthma?]. PMID- 9411446 TI - [Changes in regional CNS perfusion in obstructive sleep apnea syndrome: initial SPECT studies with injected nocturnal 99mTc-HMPAO]. AB - Some of the clinical features of obstructive sleep apnoea syndrome (OSA) are suggestive of impaired cerebral blood flow. Cerebral blood flow alterations might, for example, be responsible for headaches, which are frequent complaints in patients with OSA. Even the high frequency of ischaemic cerebral complications in patients with OSA might be caused in part by sleep apnoea-associated impairment of cerebral perfusion. Previous studies have demonstrated reduced total cerebral blood flow in patients with OSA, but regional changes of cerebral perfusion have not been studied up to now. We performed SPECT studies using 99mTc (d,l)-hexamethyl-propylenaminoxim (HMPAO) as a tracer in 14 adult patients with moderate to severe OSA (AHI > 30/h; mean AHI 59.2 +/- 4.3). The injection of the tracer took place between 2:00 and 4:00 a.m. while repeated episodes of obstructive apnoea were detected by polysomnography during stage II sleep. Data acquisition took place at 7:30 a.m. All measurements were repeated some nights later under effective treatment with nCPAP. Visual analysis showed marked frontal hyperperfusion in 5 patients. When regional perfusion indices were calculated for 32 regions of interest statistical analysis showed reduced perfusion of the left parietal region. These changes were completely reversed by effective nCPAP therapy. These data suggest that OSA is associated with reversible changes of regional cerebral perfusion. The underlying pathophysiologic mechanisms are matter of speculation so far. There might be an apnoea-associated effect of local vascular autoregulation mechanisms acting to compensate systemic blood flow alterations or blood gas changes in OSA. The observed frontal hyperperfusion might be caused by activation of the frontal lobe by repetitive cortical arousals. PMID- 9411448 TI - Tumor necrosis factor receptor-associated factor (TRAF)-1, TRAF-2, and TRAF-3 interact in vivo with the CD30 cytoplasmic domain; TRAF-2 mediates CD30-induced nuclear factor kappa B activation. PMID- 9411447 TI - [Work of breathing in differentiation of various forms of sleep-related breathing disorders]. AB - BACKGROUND: In contrast to the obstructive sleep apnoea syndrome (OSA) the obesity-hypoventilation syndrome (OHS) is characterised by persistent hypercapnia during the day and predominant hypoventilation during sleep. In this study we wanted to know whether work of breathing (WOB) in a sitting and supine position separates both groups. PATIENTS AND METHODS: OSA population: 20 men, 50.5 +/- 9.2 years, Body Mass Index (BMI: 54.1 +/- 6.9 kg/m2, pO2: 65.6 +/- 6.6 mmHg, pCO2: 40.6 +/- 3.1 mmHg, OHS-group: 14 patients, 13 men age: 53.1 +/- 9.3 years, BMI: 53.1 +/- 9.3 kg/m2, pO2: 51.8 +/- 10.5 mmHg, pCO2: 53.8 +/- 9.2 mmHg. The control group consisted of 10 normal weighted subjects. The intrathoracic pressures were assessed by an oesophageal catheter; at the same time, the minute ventilation (VE) and the breathing frequency (fb) were measured via a pneumotachygraph. The area under the pressure-volume loop was correlated to WOB. After reaching steady state VE, fb, and WOB were determined in sitting and supine position. RESULTS: In the OSA-group the apnoea index (AI) was 48.6 +/- 17.7/h and the respiratory disturbance index (RDI) was 66.3 +/- 19.4/h. The forced expiratory volume (FEV1) was 77.3 +/- 23% pred. and the vital capacity (VC) was 76.3 +/- 18.6% pred.; 7 out of 20 patients suffered from chronic bronchitis. In the OHS-group the AI was 21.5 +/- 19/h and the RDI 44.3 +/- 28.2/h. The majority of OHS patients had an airway obstruction (FEV1: 55.8 +/- 17.5% pred., VC: 58.8 +/- 12.8% pred.); 12 out of 14 patients suffered from chronic bronchitis. Compared to the OSA population WOB in the OHS group was significantly higher both in the sitting (0.67 +/- 0.28 J/I versus 1.04 +/- 0.32 J/I, p < 0.001) and supine positions (1.23 +/- 0.25 J/I versus 1.91 +/- 0.43 J/I, p < 0.001). Compared to the sitting position VE and fb did not change significantly in both groups lying supine. CONCLUSIONS: Compared to the OSA group at the same BMI the WOB of the OHS population was significantly increased in the sitting and supine position. The main reason for these findings may be the increased airway obstruction due to chronic bronchitis. Both populations did not change the breathing patterns during the different positions. PMID- 9411449 TI - [Voltage-dependent L-type calcium channels--inhibition, function and modulation]. PMID- 9411452 TI - [Pharmacy at the Univeristy of the Saarlands]. PMID- 9411450 TI - [Local and regional therapy: benefits, new findings, examples (part one)]. PMID- 9411453 TI - [New findings on endogenous regulation of glucocorticoids]. PMID- 9411451 TI - [How dangerous is genetic technology?]. PMID- 9411454 TI - [PAF-antagonists with lipid structure. 6. Alkylpropandiol lipids with acyl- and ether- structures at the C-3 position and heterocyclic head groups; synthesis, characterization and structure-activity relationship]. AB - A series of 14 PAF-analogues with simple lipid structure and heterocyclic head groups were synthesized, and the PAF-inhibitory potencies on human blood platelets was evaluated in vitro. Structure-activity relationships revealed that the PAF-antagonist activity is strongly influenced by the distance between position C-3 of the backbone and onium center and the structure of the heterocyclus. The best activity showed the 3,5-dimethylpyridinium derivative with a distance of 5 methylene groups (IC50 = 0.8 mumol/l). PMID- 9411455 TI - The Physiology and Functional Diversity of Amiloride-Sensitive Na+ Channels: A New Gene Superfamily. Proceedings of the 1997 APS Conference. Park City, Utah, October 29-November 1, 1997. Abstracts. PMID- 9411456 TI - The electronic Plant Gene Register. PMID- 9411457 TI - Bow, range, and sequential effects in absolute identification: a response-time analysis. AB - Accuracy and response time (RT) measures were obtained in the absolute identification of line length. Different groups of subjects performed the task under different experimental conditions where range and relative spacing were the main independent variables. For comparison purposes, another group of subjects performed a digit-identification task. Results were analyzed for bow, range, and sequential effects using both accuracy and RT data. A preliminary analysis of RT distributions was also performed. Several phenomena previously documented using accuracy are reproduced, while new observations are reported for RT. The results show a dissociation between RT and accuracy in that the experimental manipulations sometimes affected accuracy but not necessarily RT. PMID- 9411459 TI - Explicit speech segmentation and syllabic onset structure: developmental trends. AB - Recent applications of the hierarchical theory of the syllable to the development of explicit speech segmentation are critically examined. One particular prediction, that an initial consonant is more easily isolated when it constitutes the complete onset of a syllable than when it is part of a cluster onset, was tested on children with grade levels ranging from kindergarten to second grade. At each level, two independent groups of children worked with either CVCC (first consonant complete onset) or CCVC (part of cluster onset) syllables. First- and second-graders performed better on the CVCC than on the CCVC material in an initial consonant deletion task, but not when the task was comparison on the basis of that consonant. With the same instructions as the older children, kindergarten children performed at floor level on both tasks with both materials. However, in a new experiment in which the deletion task was presented as a puppet game, and with pretraining and selection on vowel deletion, a significantly higher level of success was achieved by the children working with the CVCC material. These results are consistent with the notion of developmental precedence of onset segmentation on phoneme segmentation. On the other hand, the results of the first and second graders show that onset superiority is not specific for the pre-reading stage. PMID- 9411458 TI - In support of hierarchy in object representations. AB - The descriptive minimum principle states that the preferred interpretation of a pattern is reflected by the simplest representation of that pattern. Such a simplest representation generally has a hierarchical structure. The pattern component represented at the highest hierarchical level is said to constitute the "superstructure" of the pattern, and pattern components represented at lower levels are said to constitute the "subordinate" structure. The primed-matching paradigm has been employed in two experiments to test whether superstructures of three-dimensional objects are perceptually more dominant than subordinate structures. In the first experiment, the test pairs consisted of two-dimensional line drawings of three-dimensional objects; each prime was a two-dimensional face of such an object, corresponding to either the superstructure or the subordinate structure. Two priming conditions were employed. In the "literal" condition, the object face was presented as it appeared in the drawing of the object (physical similarity). In the "frontal" condition, the object face was presented in the frontal-parallel plane (representational similarity). Object matching was found to be facilitated more by priming superstructures than by priming subordinate structures. In the second experiment, the order was reversed; the test pairs were composed of the object faces and the object drawings wee taken as primes. Again, there were facilitating effects for both superstructures and subordinate structures, but this time without differentiation between superstructures and subordinate structures. PMID- 9411460 TI - [Psychoanalytically oriented treatment of anorexia nervosa. Methodology-related critical review of the literature using meta-analysis methods]. AB - Psychodynamic approaches to treatment in anorexia nervosa (AN) remain influential. It is unclear, how a psychodynamic orientation influences outcome with this patient group. To our knowledge, there exists no systematic review (sensu Cochrane) of the subject. METHODS: Extensive electronic and hand searches were conduced (updated in December 1995) to identify case reports and all longitudinal and comparison studies using "psychodynamic" treatment approaches, published in English, French and German. Case reports were examined for treatment "components". Group studies meeting operationalised criteria were independently rated for methodological quality, 12 aspects of treatment and its delivery (setting, type, orientation and focus) and therapy dose and duration. Effect sizes were calculated for post treatment and follow-up. Multiple regression analysis was used to identify outcome predictors. FINDINGS: Six treatment comparison studies and seven follow-up studies met criteria. In most case reports and studies treatment included different settings and techniques and an explicit focus on the symptom. In the two studies directly comparing undifferentiated psychodynamic treatment and "treatment as usual" resp. With psychodynamically based disorder specific treatments the latter had better results. Outpatient treatment was as successful as inpatient treatment (in somewhat different populations). It was not possible to identify outcome predictors. INTERPRETATION: "Psychodynamic" treatment in AN is more pragmatic and disorder orientated than generally acknowledged; patients seem to fare better in specialized treatment programmes delivered by experienced teams; less cachectic patients can be treated as outpatients by a specialized team. The exact contribution to treatment outcome of the psychodynamic orientation remains to be delineated. There is a great need for well designed and well presented treatment outcome studies. PMID- 9411462 TI - [Psychoanalytically-based treatment of bulimia nervosa]. AB - There is a long tradition of psychoanalytically oriented treatments for in patients in Germany and a large proportion of bulimics is treated in this way, but there are very few reported controlled trials for this form of treatment. The treatment rationale for psychoanalytically orientated psychotherapy is discussed against the background of the specific psychopathology of bulimia and the principal elements of this form of treatment are presented. Recent results of a controlled study involving n = 32 Patients with a follow-up of 38 months are reported with regard to comparable evaluations. The possible healing factors via which psychoanalytically orientated treatments achieve their effects with bulimia nervosa are discussed. PMID- 9411461 TI - [Behavioral therapy, cognitive behavioral therapy and cognitive-analytic methods in treatment of anorexia]. AB - This paper describes the current state of behavioural, cognitive-behavioural and cognitive-analytical treatments of anorexia nervosa and the underlying theoretical models. Purely behavioural treatment methods have been evaluated in a number of single case studies. Although effective in terms of increasing body weight, these methods are obsolete in view of their unpleasant side-effects. Cognitive-behavioural and cognitive-analytical therapies are much more appropriate for these patients given their complex symptomatology and frequently ambivalent attitude to treatment. However, so far evaluations of these treatments are rare. The reasons for this are discussed. PMID- 9411463 TI - [Are psychotropic drugs necessary for the treatment of anorexia and bulimia nervosa?]. AB - This review summarises the results of psychopharmacological treatment studies on anorexia and, bulimia nervosa. Although several drugs have tested in patients with anorexia nervosa, the outcome of controlled studies has been disappointing. Trials of pharmacotherapy for bulimia nervosa have demonstrated that tricyclic antidepressants, monoamine oxidase inhibitors and selective serotonin reuptake inhibitors significantly reduce the frequency of binge eating and purging. In some cases, psychotherapists should accept the necessity of psychopharmacological intervention, although this does not imply a known biological cause of the eating disorder. However, the significance of antidepressant medication in the overall treatment of anorexia and bulimia nervosa remains unclear. PMID- 9411464 TI - [Comparison of controlled psycho- and pharmacotherapy studies in bulimia and anorexia nervosa]. AB - Controlled studies of psychological, pharmacological and combined treatments for bulimia and anorexia nervosa were examined. Only studies with random assignment to treatment groups were included. Outcome criteria differed for anorexia and bulimia nervosa. The bulimia nervosa studies were evaluated in a meta-analysis. The results of the abstinence and reduction rates as well as the magnitude of the effects confirm the superiority of psychological approaches to pharmacological treatment with antidepressants. The few combined treatment studies suggest the same results as for psychological treatment alone. Longer treatment duration turned out to be a positive predictor for the effects of purging and depression. As far as anorexia nervosa is concerned, there is a general and significant lack of controlled treatment studies. Since almost all of the existing psychological and pharmacological treatment studies involve additional treatment elements, a clear assignment to the treatment categories is difficult. PMID- 9411465 TI - [A German version of the Eating Disorder Inventory EDI-2]. AB - The paper presents a German version of the second revised edition of the Eating Disorder Inventory EDI-2 (Garner 1991). The EDI-2 is a self-rating inventory (self-report measure) with 91 items and 11 subscales designed for the assessment of attitudinal and behavioural dimensions relevant to anorexia and bulimia nervosa. It consists of the eight original subscales: drive for thinness, bulimia, body dissatisfaction, ineffectiveness, perfectionism, interpersonal distrust, interoception and maturity fears, and the three new subscales: asceticism, impulse regulation and social insecurity. The German EDI-2 was given to 71 patients with anorexia or bulimia nervosa, 30 patients with binge eating disorder, a control group of 186 women and a further control group of 102 men. In comparison to the female control group, patient groups showed significantly elevated means on all subscales. Item analysis revealed sufficient internal consistencies for all subscales except subscale 9 (asceticism) with Cronbachs alpha ranging from 0.58 to 0.90. Twelve of the 91 items showed poor item total scale correlations below 0.40. Factor analysis supported a six-factor-structure. Hence, the reliability and validity of the three new subscales was confirmed only partially. The use of the EDI-2 in therapy research and clinical practice is critically discussed. PMID- 9411467 TI - [Perception of a group of second grade boys and girls about AIDS]. PMID- 9411466 TI - [Permanent renal disease in Puerto Rico, morbidity and mortality trends 1970 1994]. AB - To describe the characteristics and trends of the incidence and mortality of End Stage Renal Diseases (ESRD) in Puerto Rico, a descriptive analysis of the data of all patients treated with dialysis between 1970 through 1994 was conducted. A total of 7,256 patients received dialysis treatment for ESRD in Puerto Rico. Of these, 61% were males and 39% were females. Diabetes (41.7%) and glomerulonephritis (18.3) accounted for the largest number of cases followed by circulatory problems (8.3%). Hemodialysis was the predominant treatment modality (76%). The incidence trend was significant for the predictor variable period of time after adjusting by age and gender (Poisson). The mortality trend was significant for the predictor variables, period of time, gender and age (Poisson). A substantial increment in the incidence and mortality of persons receiving dialysis was found, particularly in the diabetics, males and elderly persons. The casual explanations of these findings requires further study. PMID- 9411468 TI - [Bioethical bases in transplantation]. PMID- 9411469 TI - [What is my practice worth? Basic principles for evaluating radiology practice with the expected financial yield calculations--2]. PMID- 9411470 TI - [Steps for estimating the value of a radiology practice]. PMID- 9411471 TI - [Section 6a GOA: private health insurance negates a complicated legal status]. PMID- 9411472 TI - [No fee deduction, because..... The most interesting section 6a GOA cases from ongoing legal decisions]. PMID- 9411473 TI - [Expert consultation fees are frequently inflated]. PMID- 9411474 TI - [An integrated early detection concept in women with a genetic predisposition for breast cancer]. AB - Breast cancer is in 5% of cases due to a genetic disposition. BRCA1 and BRCA2 are by far the most common breast cancer susceptibility genes. For a woman with a genetic predisposition, the individual risk of developing breast cancer sometime in her life is between 70 and 90%. Compared to the spontaneous forms of breast cancer, woman with a genetic predisposition often develop breast cancer at a much younger age. This is why conventional screening programs on the basis of mammography alone cannot be applied without modification to this high-risk group. In this article, an integrated screening concept for women with genetic predisposition for breast cancer using breast self-examination, clinical examination, ultrasound, mammography and magnetic resonance imaging is introduced. PMID- 9411475 TI - [Digital magnification mammography in computed radiography. Initial clinical results]. AB - INTRODUCTION: The combination of direct magnification mammography and computed radiography provides an improvement in spatial resolution of storage phosphor based digital systems. A clinical study comparing conventional and digital direct magnification mammograms was performed. METHODS: 100 survey mammograms in 1.5-or 1.7-fold magnification and 50 4-fold spot magnification views were obtained with a prototype direct magnification mammography system and a storage phosphor-based digital system. An intraindividual comparison of these with previous conventional radiograms of the same patients was carried out. RESULTS: The diagnostic value of digital survey mammograms using the direct magnification technique is comparable to that of conventional radiograms of the breast, especially with regard to the identification of microcalcifications and lesions and the clinical consequences. Spot magnification views performed with this combination of techniques allowed improvement in the evaluation of microcalcifications. In 15% of cases, diagnostic procedures were adjusted accordingly. CONCLUSION: The combination of the direct magnification technique with digital storage phosphor radiography systems allows the performance of digital mammography by improving the overall spatial resolution. The diagnostic value of digital direct magnification survey mammograms was comparable to that of conventional mammograms. Digital 4-fold spot magnification views improved visualisation of the morphologic aspects of microcalcifications. PMID- 9411476 TI - [Direct radiographic magnification mammography with a new microfocus tube. Experimental studies of resolution and radiation exposure]. AB - A recently developed X-ray unit for mammography using microfocal (spot size 0.12 0.05 mm) direct magnification radiography will soon be introduced into clinical practice. Unit arrangement and tube construction have been demonstrated. Mean surface dose was measured using PMMA-phantoms. A dose reduction of up to 50% was obtained in 1.7-fold magnification mammography (full sized views) compared with conventional techniques. The image quality was almost equivalent. However, in comparison with 1.7-fold magnification mammography, a comparable dose range for 4 fold magnifications with the microfocus system was measured with a substantial gain in spatial resolution. These findings satisfy the demands of a modern mammographic unit. PMID- 9411477 TI - [Computer-assisted evaluation of mammography images. Initial clinical experiences]. AB - Preceding studies have shown that a second independent reviewer of conventional mammographies increases the detection rate of features typical for malignancy by up to 15%. METHODS: In order to test a computer-aided diagnostic (CAD) system (ImageChecker, R2 Technology, USA) for the detection of pathologic criteria in conventional mammography, 96 mammographies were retrospectively evaluated using ImageChecker. Thirty-five of these mammographies had been diagnosed as not showing pathologies, and 61 had depicted histologically confirmed malignancy. RESULTS: Detecting 41 of 61 breast malignancies, ImageChecker showed a diagnostic sensitivity of 70.5%. All malignancies accompanied by microcalcifications were identified by ImageChecker, whereas 18 cases characterized by parenchymal opacity without microcalcifications were not marked. On the average, 1.95 markers per image were set, giving a total of 187 markers in this study. 63% of all markers showed normal tissue and were thus false positive. CONCLUSIONS: Pathologic parenchymal opacities in mammography are a well-known problem for all CAD systems in use. Despite this major drawback, even now ImageChecker can provide tremendous support in routine interpretation of conventional mammographies. PMID- 9411478 TI - [Quality assurance in roentgen mammography. Comparison of recommended EUREF guidelines with relevant German regulations]. AB - The Program "Europe against Cancer" published the 2nd edition of quality assurance guidelines for breast cancer screening in June 1996. For the enforcement of these guidelines, a European network of reference centres (EUREF) is being established. Although the EUREF protocol contains guidelines for all disciplines involved in breast cancer screening, this article concentrates on the physical and technical aspect. The comparison with the German regulations (DIN Norms) demonstrates the high requirements requested by the EUREF guidelines with its tighter limits and more extensive and more frequent tests. PMID- 9411479 TI - [Vacuum punch biopsy of the breast with a stereotaxic guide. A new procedure for percutaneous diagnostic biopsy based on 120 cases]. AB - PURPOSE: To test the capabilities of vacuum core biopsy (VCB) in the diagnosis of mammographically indeterminate lesions. MATERIALS AND METHODS: 120 patients (131 lesions) were examined using VCB with 14G or 11G vacuum core. RESULTS: VCB was mostly performed because of indeterminate microcalcifications (67 cases) or soft tissue densities/architectural distortion (64 cases). 112 benign changes, 14 DCIS and 5 invasive carcinomas were found. Excellent accuracy was achieved (presently 100%), since complete excision of small lesions/areas (< or = 1 cm) or partial excision of larger lesions was possible. No relevant hematomas or infections occurred. Patients tolerated the painless procedure very well. DISCUSSION: This report confirms our previous experiences. This method promises to replace diagnostic open biopsy of indeterminate or suspicious nonpalpable lesions. PMID- 9411480 TI - [Sitting or supine stereotaxic core biopsy of the breast? A comparison based on a randomized, prospective study]. AB - OBJECTIVE/MATERIAL AND METHODS: In a prospective randomized study, the techniques of stereotactic breast biopsies in prone and sitting position were compared. Part of the data has already been published. A total of 103 women underwent stereotactic breast biopsies, either prone (n = 51; using TRC-Mammotest, Sweden) or in the sitting position (n = 52; using Stereotix 2, General Electric Medical Systems, Milwaukee, Wisconsin, USA). With the help of pre- and post-biopsy questionnaires, anxiety, pain, and subjective experience were recorded in all patients. Vasovagal reactions were scored from 0 to 2 according to their severity. All biopsy results were verified by surgery. The specificities and sensitivities for the two positions were calculated and statistically compared. RESULTS: With regard to overall tolerance no statistically significant difference between biopsies performed in the sitting or the prone position was noted. Significantly more patients (p = 0.04) in the prone position stated they would prefer premedication prior to a repeat biopsy. Three patients (prone; n = 1; sitting; n = 2) fainted during the procedure. There was no statistically significant difference between the two biopsy positions regarding sensitivity (95%) and specificity (100%). CONCLUSIONS: More attention should be paid to patient care and, especially, preintervention information. Biopsies in the prone or sitting position are equally well tolerated. Somatic reactions are not a major problem during breast biopsy. Success and validity are independent of the biopsy position. PMID- 9411481 TI - [Modern ultrasound diagnosis of the female breast. Possibilities and limits]. AB - PURPOSE: The rapid development of sonography made it seem advisable to reconsider the clinical value of breast ultrasound. Our experience with new techniques and current research will be presented. METHODOLOGY: Various indications for breast ultrasound will be discussed. New Doppler techniques extending the ability to detect low blood flow will be presented. RESULTS: The differentiation of suspicious areas seen on mammography and the detection of occult breast cancer are the major challenges in breast ultrasound. A limitation is its low capability to visualize microcalcifications. A promising approach is the development of highly sensitive, color-coded techniques for the detection of minimal blood flow. Another progress is the development of an ultrasound contrast medium that is not inactivated during first lung passage. CONCLUSION: Breast ultrasound provides complementary information to mammography. Using stable ultrasound contrast medium and highly sensitive color-coded Doppler techniques for the detection of minimal blood flow, we hope to find new criteria for the diagnosis of angioneogenesis in breast cancer. PMID- 9411482 TI - [Diagnosis of blood flow in breast tumors with increased blood pressure. New possibility in tumor field diagnosis]. AB - On the assumption that the architecture of blood vessels of malignant tumors, formed by neoangiogenesis, shows characteristics that are different from those of blood vessels of benign tumors or physiological findings, we have tried in the present study to investigate the behavior of these different vessels under increased blood pressure. Using a special stand, the same sonographic section could be stably maintained during an examination time of approx. 4 minutes. Using a new computer program, the color pixels of the employed Angio color technique were quantified and recorded as a function of the measured blood pressure. To increase blood pressure, the patient had to press a hand grip, which practically always caused a systolic blood pressure elevation of more than 15-20 mmHg. Seventy patients with sonographically detected breast tumors were examined; 54 (14 benign and 40 malignant tumors) could be included in the evaluation. We found four typical types of curves: Curve type 1 is associated with an instantaneous increase in blood flow with increased blood pressure, followed by a drop in the blood pressure, with a slow decrease in blood flow as the blood pressure drops (with 29 malignant and 3 benign tumors). Curve type 2 shows a continuous increase in blood flow-though somewhat delayed with respect to the rise in blood pressure which is also observed when the blood pressure drops (exclusively benign tumors). In curve type 3, maximum blood flow is reached after the blood pressure maximum, and then the blood flow decreases (1 benign and 3 malignant tumors). Curve type 4 features decreased blood flow in spite of increased blood pressure (3 benign and 5 malignant tumors). The described quantification method, in combination with the stand, permits for the first time analysis of a tumor under increased blood pressure as to its blood flow behavior over time in an examination using a challenge test. Here one can find two distinctive curve types (types 1 and 2) that correlate mostly with malignant (type 1) or benign (type 2) breast tumors. Should this tendency be substantiated by additional large-scale studies, it would seem that a new ultrasonic possibility for differential diagnosis has been found. PMID- 9411484 TI - [Mediastinal space-occupying lesion with reduced perfusion of the left lung. Thoracic aortic aneurysm with compression of the left pulmonary artery]. PMID- 9411483 TI - [Classification of hemodynamic changes in renal artery stenosis using cine magnetic resonance phase contrast flow measurements]. AB - PURPOSE: To evaluate the use of high-temporal resolution cine MR phase-contrast flow measurements for assessment of flow dynamics in renal artery stenosis (RAS). MATERIAL AND METHODS: In a dog model, cine MR flow measurements were validated by comparing the MR flow data to an invasive transit-time ultrasound reference technique for different degrees of RAS. Cardiac-gated MR flow curves were recorded in 56 renal arteries of 28 patients with a temporal resolution of at least 32 ms. In all cases RAS was confirmed by digital subtraction angiography (DSA). Abnormalities of flow dynamics were assessed in the calculated flow curves using the MR parameters mean flow, maximum velocity, and time to systolic maximum. RESULTS: By means of the MR blood flow parameters high-grade stenoses (> 50%, n = 23) were detected with sensitivity of 100% and specificity of 94% with reference to DSA. The overall differentiation between stenoses (n = 37) and non stenosed vessels (n = 19) revealed a sensitivity of 87% and a specificity of 100%. CONCLUSION: Analysis of cardiac-gated MR flow curves provides a non invasive method to assess the hemodynamic significance of RAS and thus allows a functional evaluation in relation to the morphologic characteristics of the stenosis. PMID- 9411485 TI - [34th Annual meeting of the Society for Pediatric Radiology. Salzburg, September 19-20, 1997]. PMID- 9411486 TI - [Amylase production by Aureobasidium pullulans in liquid and solid media]. AB - Amylase production by a strain of Aureobasidium pullulans isolated in the laboratory was evaluated in liquid media (complex and synthetic) and in solid medium (wheat bran). There was an inhibitory effect in amylase production or amylase secretion by glucose. Asparagine was the best nitrogen source for amylase production (4-6 g/l). Only chlamidospores and melanin but not, amylase activity, were obtained with ammonium sulfate. Amylase production in solid culture was higher than the production obtained in the liquid media assayed. Optimum initial moisture content in solid culture ranged between 57 and 74%. No difference was observed in amylase production between solid media inoculated with cells grown in liquid or solid media. PMID- 9411487 TI - [Dynamics of bacteria from the phyllosphere and leaves of soy (Glycine max L. Merrill) in field conditions]. AB - The surfaces of aerial plant provide a habitat for epiphitic microorganisms many of which are capable of influencing the growth of pathogens. In this study, fluctuation of bacterial population occurring in different growth stages of soybean leaves (Glycine max L Merrill) was examined using the dilution plate method. Three culture media were applied; invariably, the numbers of bacteria increased with increasing plant age. The total number of bacteria living on the detached leaves were ten times higher than leaves surfaces of soybean Asgrow 3127 (Group III). One hundred and seventy-five heterotrophic bacteria were obtained from both the phyllosphere and litter of the plant. Of these, fifty-one microorganisms (29%) were classified as members of the genus Bacillus; the remainder were mainly members of the irregular, nonsporing, Gram positive rods (coryneform bacteria). Approximately 52% of the total coryneform bacteria isolated was found to belong to the following two clusters: Arthrobacter and Corynebacterium. There were few Pseudomonas strains and 75% of the total isolated bacteria were able to grow in N-poor culture medium. PMID- 9411488 TI - [Bovine leukemia virus (BLV): prevalence in the Cuenca Lechera Mar y Sierras from 1994 to 1995]. AB - The Cuenca Lechera Mar y Sierras (CLMS) includes about 300 dairy farms located in the counties of Tandil, Balcarce, Juarez, Ayacucho, General Pueyrredon, Gonzalez Chavez and Necochea, in the province of Buenos Aires. The purpose of this investigation was to determine the prevalence of infection caused by Bovine Leukemia Virus (BLV) in the CLMS. We investigated the presence of anti-BLV antibodies in 4,203 milk samples taken from 73 dairy farms belonging to the CLMS. An indirect ELISA, which is described and evaluated in this paper, was used to test the antibodies in milk. We classified the dairy farms according to their rate of infection. The percentage of dairy farms free of infection resulted in 31.50. On the other hand, 49.40% of the dairy farms showed a figure between 1% and 15% of infected cattle; 17.80% between 16% and 30%, and the remaining 1.30% turned out more than 30% of infected cattle. If compared with data obtained in the 1979-1981 period, which showed that 95.65% of the dairy farms was BLV-free, it is clear that a dramatic progress of the BLV infection has occurred for the last 15 years. Nevertheless, the CLMS is in a privileged position so as to incorporate an inexpensive control plan to eradicate the BLV infection, as almost 1/3 of its dairy farms is still BLV-free and 49.40% still has a low rate of BLV infection. Only about 20% of the dairy farms would require costly strategies of control. PMID- 9411489 TI - [Detection of Clostridium botulinum spores in honey]. AB - A total of 177 honey samples were examined for Clostridium botulinum, 68 of which were from commercial origin, 8 from small rural producers for family consumption, and the remaining 6 from fractionizing centers in Mendoza and San Luis provinces in Argentina. C. botulinum type A was detected in two samples of rural producer origin (1.1%) by the centrifugation-dilution method. The strain was recovered from one of the samples, obtaining a spore count of 55/g of honey. Even though the positive percentage was lower than that found in other countries, honey consumption by children under one year old should be avoided in order to prevent infant botulism. PMID- 9411490 TI - [Effect of the fungicide captan on Azospirillum brasilense Cd in pure culture and associated with Setaria italica]. AB - The effect of fungicide captan on growth and nitrogenase activity of Azospirillum brasilense Cd was studied in pure cultures and in association with foxtail millet (Setaria italica) cultivar Carape under laboratory conditions. The 8 h growth in rotary shaker of A. brasilense was inhibited with 1 mg/l pure captan; however, after 4 days the differences diminished compared with the control without captan. Nitrogenase activity was affected with 10 mg/l but the differences were negligible after 48 h of growth. Root dry weight of inoculated plants was diminished by the treatment of foxtail millet cv. Carape with captan. Inoculation with A. brasilense Cd increased shoot dry weight, but differences were significant only with respect to the control but not in relation to captan treatments. PMID- 9411492 TI - [Infections with HBV virus in health care setting in Poland and their prophylaxis]. PMID- 9411491 TI - [Mycoplasmas and AIDS]. AB - AIDS is a complex illness due to HIV type 1 and 2 infection. It is characterized by an important immunodeficiency mainly caused by depletion of CD4+ T lymphocytes. The reasons for this depletion have not been sufficiently clarified yet. In 1986, Shy Ching Lo astonished the scientific community with reported evidence concerning the direct role played by mycoplasma in the etiopathology of AIDS. Since then, different theories have pointed to mycoplasma as cofactors, commensals or opportunistic agents. Although in vivo and in vitro experiments are controversial they suggest a possible mechanism that would explain the synergism between both agents: the mycoplasma belonging to normal intestinal flora could move to urethra, oropharynx or blood due to high risk sexual practice. There it would proliferate favoured by early immunological disorders related to HIV. It has been speculated that several microorganisms including mycoplasma, acting as superantigens, could induce a chronic CD4+ and CD8+ T lymphocytes activation resulting in apoptosis of the infected lymphocytes. The release of cytokines induced by mycoplasma could influence the progression of the disease. PMID- 9411493 TI - [Iatrogenic hepatitis B, non-A non-B, and C virus infections acquired in health service institutions of the Gdansk province in 1986-1995]. AB - The aim of this study is estimation of the frequency of nosocomial infections caused by HBV, NANB virus, HCV in health service institutions of Gdansk province. Relationship between medical procedures and acute viral hepatitis was examined among 4268 patients hospitalized in Clinic of Infectious Diseases and Provincial Hospital of Infectious Diseases in Gdansk, from 1986 to 1995. The analysis of results showed that 1915 (44.9%) cases of viral hepatitis caused by HBV, NANB virus, HCV were probably connected with diagnostic or therapeutic procedures. Within ten years number of HBV, NANB, HCV infections decreased but at the same time relative increase of nosocomial infections caused by those pathogens was observed. Transfusion anamnesis was noted among 129 patients (8.5% of all persons with acute viral hepatitis probably infected in hospitals). PMID- 9411494 TI - [Analysis of factors affecting HBV and HCV infections in premature children]. AB - Premature children with the very low birth weight, who need a long hospital treatment after the birth, belong to the group of the highest risk of HBV and HCV infections. The study includes 32 premature children, 14 girls and 18 boys, who were born between 26 and 35 week of pregnancy with the birth weight from 800 to 2400 g. Chronic hepatitis were found in these children; 11 children had HBV infection, 10 children had HCV infection, and both HBV and HBC infections were found in 11 children. Blood transfusions, parenteral nutrition, parenteral antibiotics, surgical treatment and other medical interventions were considered as the most important factors affecting HBV/HBC infections. All these factors should be taken into consideration in efforts to reduce the frequency of HBV and HBC infections in premature children. PMID- 9411495 TI - [Evaluation of medical supplies sterilization at health care institutions in Poland]. AB - The assessment of the condition of sterilization of medical supplies in health care settings in Poland was done. The following issues were taken into consideration: preparation of medical supplies for sterilization, methods of sterilization, including used apparatus, and the control systems for monitoring the quality of sterilization process. It was shown that the system of sterilization of medical supplies in Poland is not satisfactory. There is the need for improvement actions, especially including: the change for dry-heat sterilizers to autoclaves; the organization of the centers for sterilization services; the installation of washer-disinfectors, and the wide introduction of monitoring of the quality of sterilization processes with the use of chemical indicators and the periodic control with biological indicators. PMID- 9411497 TI - [Epidemic of ECHO type 30 virus meningitis in children from the Lodz region in the years 1995 and 1996]. AB - Two epidemics of ECHO virus type 30 meningitis were compared with the special emphasis on the course of the disease and results of the laboratory investigations. The first epidemic occurred in 1995 and the second one in 1996. Between 20 August and 20 October 1995, thirty six children, and between 11 July and 15 October 1996, ninety one children were admitted to the Department of Infectious Diseases, Medical University of Lodz. All children were 3 to 15 years old, the median age was 9.08 years in the 1995 epidemic and 8 years in the 1996 epidemic. The common symptoms were headache, fever, vomiting and nuchal rigidity. The course of both epidemic was mild, there were no complications and serious sequelae of the disease. Typical symptoms were found more frequently in patients in 1996 epidemic. Patients in 1996 had higher CSF pleocytosis and the percentage of granulocytes in the CSF than patients in 1995. From the other side, patients in the 1995 epidemic had higher fever of longer duration than patients in the 1996 epidemic. PMID- 9411496 TI - [Vaccination against hepatitis B in children with chronic hematologic diseases treated with immunosuppression]. AB - Prevention of hepatitis B infection is an important factor in the successful management of cancer and aplastic anaemia cases. Our result suggested that children with Hodgkin's disease and solid tumors vaccinated during early stage of immunosuppressive therapy are good responders to hepatitis B vaccine. Active immunisation with hepatitis B vaccine (Engerix B), was also effective in children with leukaemia after completing immunosuppressive therapy. Protective levels of antibodies remained 6 years after vaccination. Vaccination according to shortened schedule (0-10-20 days) was not effective in these children. Passive immunisation is indicated in children with chronic neoplastic haematological diseases during immunosuppressive therapy. In 6 children the lack of seroconversion after vaccination was due to immune disorders. PMID- 9411498 TI - [Can we control pertussis better? I. Changes in the epidemiology of pertussis]. AB - In many industrialized countries pertussis has been successfully controlled due to introduction of immunization programmes. However, decline trends in pertussis incidence, observed since more than 20 years, has been recently halted in individual countries, including Poland. In some industrialized countries, even absolute increase of pertussis incidence is recorded. Important changes in age distribution of pertussis patients are noted; from one side, there is an increase of pertussis incidence in infants, and a slow but continuous shift towards older age. There is an increasing body of evidence that adults may be the main reservoir of pertussis organisms and play an important role in the transmission of pertussis infection to younger children. Studies on the role of adults in transmission of pertussis infection to younger children should be undertaken in Poland. Early beginning and early completion of series of primary immunizations with DTP vaccine in infants and maintainance of a high immunization coverage with all doses of DTP vaccine specified in the immunization calendar are needed to successful control of pertussis. There is an urgent need to improve the diagnosis of pertussis in Poland, and especially the bacteriological confirmation of the diagnosis. PMID- 9411499 TI - [Can we control pertussis better? II. Old and new vaccines against pertussis]. AB - Although in many countries pertussis had been successfully controlled by the routine mass immunization in infants and children, the disease continues to cause extensive morbidity and mortality throughout the world. Whole-cell pertussis vaccine plays an important role in the control of pertussis in the world and the vaccine proved to be very successful during nearly 50 years of its use. However, the whole-cell pertussis vaccine causes an increased rate of local and general adverse events although many of these events described as caused by vaccination are occurring co-incidentally, not being related to vaccination. Results of recent clinical and field trials in the USA, Sweden, Italy, Germany and Senegal showed that acellular pertussis vaccines are effective in preventing pertussis in children and safe in infants. However, a crucial issue in more general use of acellular pertussis vaccine is its availability and its price. It may be prove too expensive for many countries, including Poland. The cost can be expected to decrease in the future, when the production capacity and the number of doses used increase. The introduction of acellular pertussis vaccine in the immunization programme in Poland will need several organizational, technological and scientific actions. PMID- 9411500 TI - [Purulent meningoencephalitis treated in the Infectious Diseases Clinic of the Silesian Medical Academy in Bytom in 1994-1995: personal observations]. AB - Among 267 patients with central nervous system infections, 43 patients (16.1%) suffered from purulent bacterial meningitis. An etiological agent was established in 15 cases (34.9%): Str. pneumoniae--9 cases, N. meningitidis--4 cases and Staph. aureus--2 cases. Most patients had severe course of the disease; lethality was 18.6%, the recovery with subsequent sequelae was noted in 11.6% cases, and 69.8% cases fully recovered. In two patients brain abscess and intracranial empyema, and persistent cerebral ischaemia were found, one of these patients died. Frequent use of antibiotics before hospitalization reduces the possibility of establishing the etiological agent. Bacterial infections of the central nervous system are still danger diseases producing high lethality and subsequent neurological sequelae. PMID- 9411501 TI - [Etiologic factors for purulent meningitis in children. Twenty years of personal observations]. AB - Etiological agents in purulent meningitis cases hospitalized during two 9 years periods: 1977-1985 and 1988-1996 were compared. While the percentage of cases caused by Neisseria meningitidis was similar in the two analyzed periods (55.7% i 66.5%, respectively), the percentage of cases caused by Streptococcus pneumoniae decreased from 33.5% to 8.4% and the percentages of cases caused by Haemophilus influenzae type b increased from 0.6% to 14.2%. The analysis of the sensitivity of isolated strains of H. influenzae type b shows the occurrence of strains resistant to ampicillin and chloramphenicol. PMID- 9411502 TI - [Nitric oxide (NO) in children with meningitis]. AB - Twenty seven children with a documented bacterial (BM)-, 73 with viral (mumps and enteroviral) meningitis and 51 controls were included. CSF white blood cell counts, glucose and protein concentrations were determined routinely. CSF nitrite and nitrate levels (the stable degradation product of NO) were determined by a modified Griess reaction. The mean +/- levels of nitrite and nitrate in CSF on admission were higher in patients with BM in comparison with controls and in children with viral meningitis. In 10 patients dexamethasone therapy was started about 10 minutes before the first antibiotic dose and given every 12 hours of 0.4 mg/kg for 2 days. At 24 to 48 hours those who received dexamethasone therapy had a significantly lower mean +/- SD CSF nitrite concentration compared with that in non-steroid treated patients. In all patients with meningitis a significant positive correlation was found between CSF nitrite and CSF granulocyte counts and also CSF protein concentration. Increased production of NO in the CSF compartment during the acute phase of BM may contribute to the inflammatory process and tissue injury. Dexamethasone therapy administered before the first parenteral antibiotic dose reduces the production of NO in the CSF during BM. PMID- 9411503 TI - [A case of cutaneous anthrax in the Lomza district]. AB - A case of anthrax is reported in 46 year old man. Cases of anthrax in animals and human beings are rare in Poland and therefore the diagnosis of the disease can be difficult. PMID- 9411506 TI - [Epidemiologic studies of oral mucosa changes occurring in children, adolescents, and adults 13-24 years of age in Warsaw]. AB - The aim of the study was the analysis of RAS occurrence in the population and a comparison to other oral mucosa lesions as well as an analysis of the influence of various factors on RAS occurrence: age, social status of the subject's, parents' education, CPITN index. In the school year 1994/95 a questionnaire concerning the frequency of RAS was distributed to children and adolescents (13 18y) from two Warsaw schools and a group of soldiers of the Polish Army serving in Warsaw. Additionally, all study participants were checked towards RAS by the use of the clinical methods according to WHO methodological guidelines. The periodontal state was measured with the CPITN index described by Ainamo. The following diseases were identified: RAS, leukoplakia, lichen planus, herpes simplex and tongue lesions. The most common declared diseases of oral mucosa was RAS, which usually occurred to patients of age below 18. Clinically the most often observed diseases were oral leukoplakia in soldiers' group and RAS in both students' groups. RAS occurs more often in students, whose parents have higher education compared with persons whose parents have primary education. Parental history of disease (cases of RAS among family members) predisposes to occurrence of RAS. Stress and exhaustion are the supporting factors of RAS incidence. Furthermore, clinical periodontal status and treatment needs may also cause the increase of RAS incidence. PMID- 9411504 TI - [The battle against acute infections in Poland between the two world wars of the twentieth century, 1918-1939]. AB - In the health policy of the Polish State during the period between the two World Wars, the fight against acute and chronic infectious diseases was considered as a great priority. Besides legislative activities, the most important task in this policy, was the organization of the institutional forms of medical care and permanent preventive activities, as well as the formation of the people's health awareness and responsibility. Those activities resulted in, among others, the reduction of morbidity and mortality due to acute infections and social diseases. The progress in the health care was the result of a consistent cooperation between all medical care sectors, medical associations, social and scientific institutions and the essential financial support from the international organizations. PMID- 9411507 TI - [Comparative evaluation of the frequency of congenital defects in newborns from the provinces of Walbrzych, Piotrkow Trybunalski and Suwalki]. AB - The aim of the study was the assessment of congenital defect frequency in 11,869 neonates born between December 1994 and July 1995 in three provinces of Poland: Walbrzych, Piotrkow Trybunalski and Suwalki. The percentage of neonates with congenital defects ranged from 1.9% in a typically industrial area (Walbrzych) to 1.2% in a mainly rural area (Suwalki), and 1.1% in a rather industrial area (Piotrkow Trybunalski). The majority of single defects consisted of anomalies of limbs and musculoskeletal deformities. Smoking habits and professional activities of mothers did not exert a clear influence on the frequency of birth defects. Only among newborns from the province of Piotrkow Trybunalski who were born by young mothers (< 20 years of age) we observed a higher frequency of congenital defects in comparison with older mothers of 20-34 years old group. PMID- 9411505 TI - [Respiratory system reactions in children with allergies exposed to air pollution at home. Epidemiologic studies in Cracow]. AB - The purpose of the paper was to check the hypothesis whether the domestic air quality together with level of outdoor air pollution increases the risk of allergy and whether allergy determines the occurrence of respiratory symptoms. The results of the study have shown that chronic cough was related mainly to allergy, however, chronic phlegm was linked with air quality. Wheezing independent from respiratory infections was associated in the strongest way with allergy (RR = 7.7; 4.7-12.7) and much weaker related to air quality score (RR = 1.3; 1.1-1.5). The occurrence of attacks of breathlessness was confirmed only in allergic children. Allergic rhinitis, similarly to wheezing, was related stronger to allergy (RR = 3.3; 2.5-4.4) than to air quality score (RR = 1; 1.0-1.2). Pulmonary volumes (FVC, FEV1) were not associated with air quality score, but the lower indices of FEV1/FVC and FEF25 - 75% were observed in those who were exposed to poor air quality. The results are in favor of the hypothesis, that air pollutants bring about spasm of smaller airways and it is independent from respiratory symptoms. PMID- 9411508 TI - [Snake bites]. AB - Vipera berus is the only viper i.e. venomous (poisonous) snake that is found in Poland. Snakebites occur rarely und usually follow attempts of catching the viper or accidental treading on it. Appropriate treatment consists of administering specific antivenom serum (antivenin). Four cases of snakebites hospitalized in 1989-1995 in the Department of Infectious Diseases of the University Medical School in Bialystok are reported. PMID- 9411509 TI - [European Congress of Radiology (ECR), European Association of Radiology (EAR), European Society of Radiology (ESR): European Radiology between the past and the future]. PMID- 9411510 TI - [Turbo STIR sequence: optimization and comparison with conventional STIR sequence in bone diseases]. AB - INTRODUCTION: The most common fat-suppressed sequence used to study skeletal conditions is the STIR sequence which has shown high sensitivity in the detection of skeletal lesions and whose main drawback is its long acquisition time. Currently, Turbo-STIR (T-STIR) sequences can shorten the acquisition time. The purpose of this study was therefore to compare the conventional STIR sequence with the new T-STIR sequence in the study of skeletal conditions to compare their diagnostic yield. MATERIAL AND METHODS: Twenty patients with different types of skeletal lesions were examined. MR examinations were performed with a Philips Gyroscan S15/ACS II unit (1.5 T). All the patients underwent a STIR sequence (TR/TE = 1500/20, TI = 180 ms, matrix = 204 x 256, NEX = 2, slice thickness = 5 mm, acquisition time = 9 min 24 s) and a T-STIR sequence (TR/TE = 1500/20, TI = 180 ms, matrix = 204 x 256, NEX = 2, slice thickness = 5 mm, TFL = 3, acquisition time = 3 min 33 s). The images were evaluated by measuring both quantitative parameters--percent contrast (%C), contrast to noise ratio (C/N), signal to noise ratio (S/N)--and qualitative parameters--lesion conspicuity, margins and extension, motion artifacts, image quality. RESULTS: The only statistically significant difference between the two sequences was image quality, which was superior in the conventional STIR sequence (p < .05). No statistically significant difference was demonstrated with the quantitative evaluation. DISCUSSION: In this study, T-STIR sequences were performed with low-high acquisition profile to acquire an actual echo time of 20 ms which permits to obtain optimal S/N with good spatial resolution. Therefore, T-STIR sequences with low-high acquisition profile provides better results than T-STIR sequences with linear acquisition profile which permits to obtain an actual echo time of 40 ms. CONCLUSION: This work shows that T-STIR sequences can replace conventional STIR sequences in the study of skeletal conditions reducing the acquisition time by 60%. This result can be obtained only by an accurate optimization of acquisition parameters. PMID- 9411511 TI - [Role of computerized tomography in the diagnosis of bone disease in multiple myeloma]. AB - PURPOSE: To assess the role of CT in the diagnosis and management of multiple myeloma (MM) and to investigate if CT findings can influence the clinical approach, prognosis and treatment. STUDY DESIGN AND PATIENTS: We reviewed the findings relative to 273 MM patients submitted to CT June, 1994, to December, 1996. The patients were 143 men and 130 women (mean age: 65 years): 143 were stage I, 38 stage II and 92 stage III according to Durie and Salmon's clinical classification. All patients were submitted to blood tests, spinal radiography and CT, the latter with serial 5-mm scans on several vertebral bodies. The CT unit was a Philips Tomoscan SR 7000. RESULTS: CT showed lysis foci in some vertebral bodies (4 cases) where conventional radiography had shown only aspecific osteopenia. CT also depicted vertebral arch and process involvement in 3 cases with the vertebral pedicle sign. Moreover, CT proved superior to radiography in showing the spread of myelomatous masses into the soft tissues in a case with solitary permeative lesion in the left pubic bone, which facilitated subsequent biopsy. As for extraosseous localizations, CT demonstrated thoracic soft tissue (1 woman) and pelvic (1 man) involvement by myelomatous masses penetrating into surrounding tissues. In our series, only a case of osteosclerotic bone myeloma was observed in the pelvis, associated with lytic abnormalities. DISCUSSION AND CONCLUSIONS: The role of CT in the diagnosis and management of MM has not been assessed, because this technique demonstrates tumor extent more accurately than radiography but CT findings do not seem to improve the clinical approach and therapeutic management of the disease. Nevertheless, we recommend CT for some myelomatous conditions, namely: a) in the patients with focal bone pain but normal skeletal radiographs; b) in the patients with M protein, bone marrow plasmocytosis and back pain, but with an inconclusive MM diagnosis; c) to assess bone spread in the regions which are anatomically complex or difficult to study with radiography and to depict soft tissue involvement; d) for bone biopsy. PMID- 9411512 TI - [Biomechanics and semiology of transient lateral patellar dislocation: reliability of magnetic resonance diagnosis]. AB - Transient patellar dislocation is a rare finding. It is usually due to direct trauma in mediolateral direction or to indirect trauma related to violent quadriceps muscle contraction. The clinical diagnosis of transient forms is usually rather difficult because the associated signs (hemarthros and pain at the joint and/or at the vastus medialis insertion) are not specific. Conventional radiography and CT are very accurate in bone studies, but exhibit major limitations in the detection of capsular injuries with intramedullary and/or cartilaginous bone involvement. We investigated MR diagnostic reliability in the study of the injuries caused by transient patellar dislocation. 526 MR examinations of the knee were reviewed retrospectively; fifteen of them were positive for transient patellar dislocation. We used a .5 T superconductive MR unit with a circumferential extremity coil. Axial, coronal and sagittal T1 weighted spin echo, T2*-weighted gradient echo and axial STIR images were acquired. The following criteria were considered specific for the diagnosis of former lateral patellar dislocation: retinacular changes; patella site; signal intensity changes in femoral and patellar intramedullary bone; joint effusion; cartilage and subchondral bone changes. All the patients with transient patellar dislocation had hemarthros with inner retinaculum involvement. The femoropatellar joint was incongruous in 115 cases; patellar subluxation was external in 8/11 patients and lateral in the remaining 3 patients. Medullary bruises were found in 8/15 cases and cartilage injuries in 13/15. Finally, other joint components, such as the posterior horn of the medial meniscus or the anterior cruciate ligament were involved in 6/13 cases. Arthroscopy was performed in 6 patients with associated injuries and confirmed all MR findings (100% agreement). To conclude, in our experience MRI was a very reliable tool to study the injuries due to transient patellar dislocation because it showed specific changes and possible associated traumas, which helps choose the most suitable treatment. PMID- 9411513 TI - [Niobium filtration in dental radiology. Effects on image quality and on dosage]. AB - The necessity of reducing the radiation dose to the patient in diagnostic radiology according to the ALARA guideline established by the ICRP has stimulated the research on additional filtration systems capable of removing the low-energy photons increasing the dose and worsening image quality. Very few literature studies deal with the effects of niobium filtration on image quality in dental radiography with the use of modulation transfer function (MTF) and square wave response function (SWRF). Only one study has considered those effects measuring dose absorption in an anthropomorphic phantom. THE AIMS OF OUR STUDY WERE: 1) to study the effects of a 30 microns additional niobium filter on image quality using the SWRF; 2) to compare the doses absorbed in vivo during a complete radiographic survey of the mouth, both with and without niobium filtration. Qualitative studies led us to conclude that niobium filtration does not significantly worsen radiographic image quality. The following doses were measured in the exposures with niobium filtration: 1678 microGy to 6000 microGy (intraoral doses) and 75 microGy to 3643 microGy (skin doses). The comparison with the doses measured during the exposures made with conventional filtration indicates that dose reduction is not significantly advantageous relative to risk reduction. In conclusion, additional niobium filtration is not advisable in dental radiology, also because of the filter cost and of the increased wear of the unit. PMID- 9411514 TI - [Fast spin echo and fast fluid attenuated inversion recovery sequences in multiple sclerosis]. AB - Fast spin echo (FSE) and fast fluid attenuated inversion recovery (fast-FLAIR) MR sequences were compared with conventional spin echo (CSE) in quantitating multiple sclerosis (MS) lesion burden. For each sequence, the total number and volume of MS lesions were calculated in 38 remitting MS patients using a semiautomated lesion detection program. CSE, FSE and fast-FLAIR images were reported on randomly and at different times by two expert observers. Interobserver differences, the time needed to quantitate MS lesions and lesion signal intensity (contrast-to-noise ratio and overall contrast) were considered. The lesions were classified by site into infratentorial, white matter and cortical/subcortical. A total of 2970 lesions with a volume of 961.7 cm3 was calculated on CSE images. FSE images depicted fewer (16.6%; p < .005) and smaller (24.9%; p < .0001) lesions and the differences were statistically significant. Despite an overall nonsignificant reduction for fast-FLAIR images (.5% and 4.8% for lesion number and volume, respectively), significantly lower values (lesion number: p < .01; volume: p < .04) were observed for infratentorial lesions, while significantly higher values were seen for cortical/subcortical lesions (lesion number: p < .01; volume: p < .02). A higher lesion/white matter contrast (p < .002), a significant time saving for lesion burden quantitation (p < .05) and very low interobserver variability were found in favor of fast-FLAIR. Our data suggest that, despite the limitations regarding infratentorial lesions, fast FLAIR sequences are indicated in MS studies because of their good identification of cortical/subcortical lesions, almost complete interobserver agreement, higher contrast-to-noise ratio and the limited time needed for semiautomated quantitation. PMID- 9411515 TI - [Role of computerized tomography in the study of pulmonary tuberculosis]. AB - We carried out a retrospective study on 4 years' work on a series of patients with proved active tuberculosis examined in the Departments of Bronchopneumology and Infectious Diseases of the Arezzo Civil Hospital. The population was heterogeneous from the immunologic viewpoint. We compared the effectiveness of Computed Tomography (CT) with that of conventional chest radiography (CR) in the diagnosis and prognosis of tuberculosis. Thirty-eight patients were examined and subdivided into three groups according to their normal/altered immunologic state: 24 of them were thus considered immunocompetent and 9 immunodeficient; 5 patients were immigrants, with different immunologic susceptibility from European men. All patients underwent chest CR in double projection and conventional CT studies of the chest; 16 patients were also submitted to high resolution CT (HRCT). Consolidation was the most frequent parenchymal lesion in immunocompetent patients (10 of 24 cases); it was often associated with cavitation (9 of 10 cases), with no clear topographic prevalence in the upper lobes. Pleural effusion, as the only sign of disease and with empyema features, was found in 10 immunocompetent patients. No predominant pattern was found in the other two groups, even though intrathoracic adenopathies were relatively more common in the immigrants (4 cases) while micronodular images (5 cases) and cavitations (5 cases) were more common in the immunodeficient subjects. Finally, chest CR always depicted the signs of pneumopathy in progress. Conventional CT was more accurate in the spatial definition of pleuroparenchymal lesions and demonstrated more nodular lesions (6 cases with CR versus 10 cases with CT) and/or infiltrating lesions (6 cases with CR versus 15 with CT) which were often masked by scars, by massive pleural effusions or by calcific fibrothorax. CT also demonstrated adenopathic mediastinal involvement (7 cases with CR versus 11 with CT), showed empyemic pleural collections and confirmed fistulas in calcific fibrothorax subjects. Moreover, HRCT depicted centrilobular micronodular images which are often ill-defined and poorly or not recognizable at CR but are signs of tuberculous infection spread along the distal airways. Posttreatment follow-up confirmed clinical recovery showing markedly and progressively fewer small nodular densities. PMID- 9411516 TI - [Twenty-six consecutive patients with acute superior mesenteric infarction. Comparison of conventional radiology, ultrasonography, and computerized tomography]. AB - Ischemic bowel disease is a rare disorder whose incidence is increasing as the mean age of the population increases. Diagnosis by clinical, laboratory and radiologic means is often difficult, and delay in definitive therapy results in substantial morbidity and mortality. A series of 26 consecutive patients, with proved acute superior mesenteric ischemia, was retrospectively reviewed: the authors report the diagnostic methods performed preoperatively, the site and the cause of infarction and the time passed between the first radiograph ans surgery. Plain abdominal radiographs were performed in 25 of 26 patients, screening abdominal US in 23 cases and CT in 19 cases. All radiological examinations were retrospectively reviewed by three authors, independently, to recognize the different signs of infarction. On plain abdominal films, the findings warranting a presumptive diagnosis of bowel infarction were air-fluid levels (84% of cases), dilated bowel loops (48%), thickened and unchanging loops (20%), gastric distension and gasless abdomen (12%), small bowel pseudo-obstruction (8%). Screening abdominal US demonstrated intraperitoneal free fluid (26%) and dilated bowel loops (22%). Abdominal CT showed air-fluid levels (79%), dilated loops and free intraperitoneal fluid (47%), intramural gas and thickened bowel loops (36.8%), engorgement of the mesenteric vessels (31%), mesenteric-portal gas, mesenteric thrombus and marked reduction in the volume of gas in the small bowel (10.5%) and paper-thin bowel loops (5%). The authors conclude that air-fluid levels, dilated loops and intraperitoneal free fluid are the most frequent findings, even though they are not specific. While abdominal plain film and screening ultrasonography can be negative, CT detects at least one abnormal finding and at least three abnormal findings in 73% of cases. PMID- 9411518 TI - [Comparison of computerized tomography and magnetic resonance with fast sequences with and without paramagnetic contrast media in the assessment of liver metastasis: qualitative and quantitative analysis]. AB - We compared unenhanced and contrast-enhanced fast MRI and CT in the detection of liver metastases. Eleven patients with single or multiple hepatic lesions (42 in all) were submitted to CT and MR studies; T1- and T2-weighted TSE, T2-weighted TSE with fat suppression, unenhanced breath-hold TFE and early or delayed enhanced breath-hold TFE images were acquired with a 1.5 T super-conductive magnet (Philips NT). The quantitative analysis of all MR images was performed for contrast/noise ratio (CNR) and number of detected lesions; MR and CT images were also compared qualitatively for lesion conspicuity, anatomical structure identification and artifacts. The results were compared with Student's t test. Early enhanced breath-hold TFE was statistically superior to T1-weighted TSE (p = .0009), T2-weighted TSE (p = .01) and CT (p = .0004) for lesion conspicuity and to T1-weighted TSE, T2-weighted TSE, unenhanced TFE (p = .0001) and CT (p = .01) for anatomical structure identification. CT was superior to T1- and T2-weighted TSE (p = .0001) and unenhanced TFE (p = .004) for the lack of artifacts. Fat suppressed T2-weighted TSE images had a statistically higher CNR than T2-weighted TSE (p = .02), T1-weighted TSE (p = .0006) and unenhanced and delayed TFE sequences (p = .007; p = .0001, respectively). To conclude, MRI appears superior to CT in the detection of liver metastases; the examination should include early enhanced breath-hold T1-weighted TFE and T2-weighted fat-suppressed TSE images. PMID- 9411517 TI - [Computerized tomography assessment of complications secondary to abdominal aortic prosthesis]. AB - PURPOSE: CT has gained an important role in the diagnosis of the complications of prosthetic surgery of the abdominal aorta: the importance of such complications comes from their frequency, which is proportional to the increasing number of interventions, and their severity. We investigated the CT patterns of the most frequent complications. MATERIALS AND METHODS: 24 patients referred for strongly suspected postoperative complications were examined in 2 years: fever and leukocytosis (20 cases) and progressive anemia (4 cases) were the most frequent findings. The operation had been performed 7 +/- 12 weeks before (2 patients were excluded because surgery dated less than 3 weeks). RESULTS: 14 patients had infective complications: thickening (57%) and inhomogeneity (43%) of the periprosthetic wrap and ectopic gas bubble (78%) were the most frequent CT findings. We also observed 2 periprosthetic hematomas, 1 aneurysm relapse and 1 prosthetic graft rupture. CONCLUSIONS: In conclusion, CT confirmed its important role in the study of the complications of prosthetic aortic surgery, despite its known poor specificity in the demonstration of the aorta in the first 2-3 months postoperatively, in the initial stages of infection and in the diagnosis of aorto enteric fistulas. PMID- 9411519 TI - [Diagnostic imaging in the selection of candidates to orthotopic transplantation of the liver. Experience at a hospital lacking a transplantation department]. AB - We report the experience of our general hospital in selecting the patients for orthotopic liver transplantation (OLT). Fifty-one patients with cirrhosis were examined and 20 of them submitted to OLT from August, 1992, to November, 1995. For liver studies, the 20 transplant recipients were examined with US and plain and dynamic CT; 15/20 were submitted to CTAP, 10/20 to Lipiodol CT and 17/20 to angiography. The accuracy of these techniques in HCC detection was assessed by correlation with resected whole livers. The accuracy of duplex Doppler and color flow Doppler for portal and/or mesenteric vein thrombosis was evaluated by correlation with resected livers, CT and angiographic findings. Pathologic examinations diagnosed HCC in 5/20 transplant recipients: 2 lesions (1.5 cm and 2 cm; 2 cm and 3.5 cm) were found in 2 resected specimens (total hepatectomy) and 1 lesion was found in 3 cases (2.5 cm, 1.5 cm, 1 cm). The sensitivity of US, plain and dynamic CT in identifying HCC patients was 20%; US and CT specificity rates were 100% and 87%, respectively. CTAP sensitivity was 75% and the sensitivity of Lipiodol CT and angiography was 100%. Therefore, in our series, US was poorly sensitive in the detection of liver cancers, which may depend on the small number of patients, lesion size (< or = 3.5 cm) and the radiologists ignoring clinical and laboratory data on purpose. Nevertheless, the patients with a single HCC not exceeding 5 cm phi or with no more than 3 tumors, none of them exceeding 3 cm phi, are generally considered eligible for transplantation: therefore, our patients chosen for OLT on the basis of US and CT findings were actually eligible for transplantation in spite of US and CT false negative results. At US, the portal vein had an average caliber of 13.5 +/- 2.5 mm in 21/51 patients; the average caliber of the common hepatic artery was 6 +/- 1.5 mm in 49/51 patients; average spleen length was 174 +/- 38 mm. US showed ascites in 28/51 cases. In conclusion, considering also the long stand-by list for OLT, the first selection of transplant candidates could be performed with US and color flow Doppler, plain and dynamic CT. The patients who are ruled out as candidates for OLT on the basis of the findings of these imaging techniques and of clinical and laboratory findings are submitted to no further examination and referred to the transplantation unit. Otherwise, if conventional and color flow Doppler US and conventional CT are not enough to exclude a patient from OLT, the subject is submitted to more invasive (angiography, CTAP, Lipiodol CT) or less widespread (spiral CT, MRI) techniques. PMID- 9411520 TI - [Intraluminal ultrasonography of the biliary ducts]. AB - We report our 4 years' experience with intraluminal US of the biliary ducts performed on 24 patients via a biliary transhepatic-duodenal drainage catheter or a surgically inserted T tube (2 cases). The final diagnosis was a malignant disease in 16 patients and a benign disease in 8 patients. We used an Intra Vascular Ultra Sound (Diasonics) unit with a 20 MHz transducer inserted into a 9 F catheter with closed tip, which in turn was pushed through the biliary ducts in a 12 F sheathed introducer which provides a safety metal guide wire in the duodenum. US examinations lasted only a few minutes and caused no additional patient discomfort; no complications followed. Intraluminal US confirmed the results of other clinical and instrumental tests in 18/24 patients and improved the diagnostic yield in assessing lesion nature and operability in 6 cases (25%). Its cost is high, but the Intra Vascular Ultra Sound includes a 3.5 MHz and a 7 MHz probes and can therefore be used for all the other US examinations needed in interventional radiology rooms and in the study of the biliary ducts. To decrease the cost, we used the Intra Vascular Ultra Sound in less than 10% of the patients treated for biliary obstruction in the same period of time, namely those with questionable diagnosis and operability. We conclude that intraluminal US of the biliary ducts is a useful tool in the centers using interventional radiology to treat many obstructive jaundice patients by a transhepatic approach. PMID- 9411521 TI - [Role of fine needle biopsy guided with computerized tomography in peripheral lung nodules with a diameter lower than 2 centimeters]. AB - We investigated the role of CT in guiding fine-needle biopsy (FNB) in small (< 2 cm phi) lung lesions, reviewing 2109 CT-guided biopsies of peripheral lung nodules performed 1983 to 1993. "Peripheral" refers to non intrabronchial lesions, independent of the nodule position in the chest. Two hundred lesions < 2 cm phi were selected, 52 of them with complete clinical and surgical follow-up. This group was compared with 195 lung lesions > 2 cm phi for CT-guided FNB sensitivity, specificity and diagnostic accuracy. Cytologic specimens were positive for malignancy in 40/52 cases (76.9%) and negative in 12 (23.1%). Clinical-surgical follow-up confirmed all the positive cytologic findings (40 cases), while 2 of the extant 12 cases were false negatives (95.20% sensitivity, 100% specificity and 96.15% diagnostic accuracy). 158 of 195 nodules > 2 cm phi and with clinical-surgical follow-up (81.3%) were positive for malignancy and 34 were negative (17.43%) (98.75% sensitivity, 97.14% specificity and 98.46% diagnostic accuracy). Pneumothorax occurred in 33 patients (16.5%) with nodules < 2 cm phi as a complication of biopsy. Hemoptysis occurred in 15 patients (7.5%). In the whole group of lung lesions, the complications were pneumothorax in 80 patients (4.1%), hemoptysis in 199 (10.4%) and hypotensive vagal crises in 92 patients (4.8%). We conclude that CT is the best guidance for FNB also in small lung nodules, providing high sensitivity and specificity and low complication rates. Finally, the result of this procedure depends on the operator's skills. PMID- 9411522 TI - [Results of flow-controlled antiblastic perfusions (stop-flow technique) carried out with percutaneous technique: 30-month experience]. AB - PURPOSE: We report the morphological and clinical results in a series of patients with advanced thoracopulmonary, abdominal, pelvic and lower limb tumors treated with stop-flow perfusion (SFP). MATERIAL AND METHODS: We performed 77 SFPs with the percutaneous angiographic technique in 55 patients (25 women and 30 men; mean age: 53 years). Eighteen thoracic perfusions with aortocaval block (ASI) were performed in 11 patients, 33 abdominal hypoxic perfusions (HAP) in 26 patients, 17 hypoxic pelvic perfusion (HPP) in 11 patients and 9 hypoxic lower limb perfusions (HILP) in 7 patients. 42/77 procedures were followed by hemofiltration. RESULTS: No technical complications were observed. Twenty-eight patients in our series are still alive (mean follow-up: 14 months) and 23 have died (mean survival: 8 months), 20/23 of them (87%) for disease progression. Three of 77 patients (3.8%) died within 7 days of the procedure (2 AS, 1 HAP). At CT or MR follow-up, responses > 50% were observed in 56% of the procedures and clinical CR was achieved in 53/77 patients (69%). In the subgroups classified by procedure, positive responses were observed in 56, 48, 59 and 78%, respectively for ASI, HAP, HPP and HLP. Clinical CR was observed in 67, 67, 71 and 78%, respectively. The death rate for disease progression relative to the overall death rate was 100, 86, 75 and 100%. Hematologic toxicity according to WHO criteria (mean: 2) was observed in 77% of the whole of procedures (59/77). Statistical analysis showed no relationship between morphological responses and type of antiblastic drug or previous antiblastic treatments. CONCLUSIONS: SF procedures permit the effective control of many advanced tumors which cannot be treated otherwise, with a high rate of positive morphological and of complete clinical responses. The best results were obtained in hypoxic perfusion of the lower limb. The results were not correlated with previous antiblastic treatments. However, the high rate of sequels and the low hematologic tolerance of those procedures must be emphasized. PMID- 9411524 TI - [Server World-Wide Web on the Internet for the provision of clinical cases and digital radiologic images for training and continuing education in radiology]. AB - The Internet, as a global computer network, provides opportunities to make available multimedia educational materials, such as teaching files and image databases, that can be accessed using "World-Wide Web" client browser to provide continuing medical education. Since August, 1995, at the Institute of Radiology University of Palermo, we developed a World-Wide Web server on the Internet to provide a collection of interactive radiology educational resources such as teaching files and image database for continuing medical education in radiology. Our server is based on a UNIX workstation connected to the Internet via our campus Ethernet network and reachable at the uniform resource locator (URL) address: http:/(/)mbox.unipa.it/approximately radpa/ radpa.html. Digital CT and MR images for teaching files and image database are downloaded through an Ethernet local area network from a GE Advantage Windows workstation. US images will be acquired on-line through a video digitizing board. Radiographs will be digitized by means of a Charge Coupled Device (CCD) scanner. To set up teaching files, image database and all other documents, we use the standard "HyperText Markup Language" (HTML) to edit the documents, and the Graphics Interchange Format (GIF) or Joint Photographic Expert Group (JPEG) format to store the images. Nine teaching files are presently available on the server, together with 49 images in the database, a list of international radiological servers, a section devoted to the museum of radiology hosted by our Institute, the electronic version of the Journal Eido Electa. In the first 12 months of public access through the Internet, 12,280 users accessed the server worldwide: 45% of them to retrieve teaching files; 35% to retrieve images from the database; the remaining 20% to retrieve other documents. Placing teaching files and image database on a World-Wide Web server makes these cases more available to residents and radiologists to provide continuing medical education in radiology. PMID- 9411523 TI - [Computerized tomography-guided neurolytic block of the splanchnic nerve]. AB - CT-guided neurolytic splanchnic nerve block is a technique for relieving abdominal cancer pain; the goal is the alcoholic neurolytic interruption of the sensitive structures in retroperitoneal space. CT yields accurate anatomical detailing and the course for needle placement and alcohol spread. January, 1993, to July, 1996, twenty-one bilateral splanchnic nerve blocks were performed through the posterior access. Forty-eight hours after alcoholization, 14 patients (66%) had complete pain regression; 52% of the patients needed no analgesics for 6 to 54 days and only 9 patients (42%) needed another low opioid therapy. Complications included hypotension and diarrhea in all cases. One had a cardiac arrest and died 8 days after the procedure. There were no other complications. The whole procedure usually lasted 60 min (range: 45 to 90 min). Splanchnic nerve neurolysis is a useful treatment in the patients with severe chronic abdominal pain. It is used as a second line treatment when large lesions change celiac anatomy and complicate the percutaneous block of the celiac plexus. PMID- 9411525 TI - [Ultrasonographic diagnosis of acute appendicitis: current indications and limitations of the method]. PMID- 9411527 TI - [Central neurocytoma: diagnosis and treatment. Report of a case]. PMID- 9411526 TI - [Prognostic factors in the treatment of carcinoma of the cervix uteri in IB-IIA stage. Retrospective study]. AB - January, 1977, to December, 1990, 311 patients with stage IB-IIA cervix carcinoma were treated at the Radiotherapy Department of the University of Turin. The distribution by stage was: 232 T1b (74.6%) and 79 T2a (25.4%). One hundred and eighty-nine patients (77% T1b-23% T2a) underwent preoperative radiotherapy, 63 patients (58% T1b-42% T2a) radiotherapy alone and 59 (85% T1b-15% T2a) postoperative radiotherapy. The first group of patients was treated according to three treatment protocols, based on different surgical procedures. During the median follow-up period of 86 months (82.6% of the patients underwent a minimum 3 year follow-up), 55 locoregional relapses (17%) and 21 extrapelvic metastases (7%) were observed. The 5-year NED survival rate and local control was 80%. The prognostic factors which significantly influenced survival in the univariate analysis, were: disease stage (p < .01), age (p < .05), the period between first symptom and therapy (p = .01), treatment protocols (radiotherapy combined with surgery versus radiation therapy alone: p < .05), residual disease after brachytherapy (p < .01), nodal status (p < .00001). In the radiotherapy alone group, the total dose influence on survival was not statistically significant (p = .12). In our series, histology and surgical procedures did not seem to influence prognosis. In the multivariate analysis, treatment protocol, age and residual disease after brachytherapy did not influence the prognosis, whereas the total dose of radiotherapy, pain (as first symptom) and Wertheim-Meigs surgery approach are prognostic factors. Severe late-effects were 17: 13 in the radiotherapy plus surgery protocol and 4 in the radiotherapy alone protocol. The incidence of major complications seems to correlate with surgical impact. PMID- 9411528 TI - [Osteosarcomatosis with pleural involvement. Report of a case]. PMID- 9411529 TI - [Synovial osteochondromatosis of the shoulder. Report of a case]. PMID- 9411530 TI - [Volvulus of the splenic flexure of the colon. Report of a case]. PMID- 9411531 TI - [Adrenal scintigraphy with Indium-111 octreotide in a case of malignant pheochromocytoma: comparison of the planar and tomographic technique (SPECT)]. PMID- 9411532 TI - [Tomodensitometry findings in a case of ileo-ileal invagination caused by intestinal myxoid liposarcoma]. PMID- 9411534 TI - [Multiple focal hepatic lesions caused by hepato-portal sclerosis. Report of a case]. PMID- 9411533 TI - [Intestinal obstruction caused by phytobezoar: computerized tomography findings. Report of 3 cases]. PMID- 9411535 TI - [Lipomatous tumor of the liver. Report of 2 cases]. PMID- 9411536 TI - [Consumption coagulopathy associated with splenic hemangiomatosis: report of a case studied with ultrasonography and computerized tomography]. PMID- 9411537 TI - ["First intention" insertion of the Palmaz stent on postoperative restenosis of the internal carotid artery. Report of a case]. PMID- 9411538 TI - ["Snow storm" ultrasonography pattern in multiple organs in a patient with AIDS and disseminated Mycobacterium avium intracellulare infection]. PMID- 9411539 TI - [Ultrasonographic features, with color Doppler, with computerized tomography and angiography in a case of abdominal Castleman's disease]. PMID- 9411540 TI - [Castleman's disease. Report of a case]. PMID- 9411541 TI - [Pyogenic liver abscess: current status]. PMID- 9411542 TI - [Comorbidity, hospitalization, and drug use for non-exacerbated chronic disease in the elderly population]. AB - OBJECTIVES: To evaluate the prevalence of comorbidity among elderly hospitalized patients and its influence on discharge diagnosis and medication due to non exacerbated chronic disease (NECD). To evaluate the impact of hospital admission on the use of drugs due to NECD since admission to the month of discharge. METHODS: A study was made of 85 patients aged 65 years or older collected during two consecutive months. The study protocol consisted of a questionnaire on comorbidity, study of drug consume, discharge diagnosis and follow-up for one month post discharge. RESULTS: Patients had a mean of 6.4 chronic diseases; significant differences were observed regarding discharge report (mean: 2.1). The number of drugs due to NECD prior to admission (mean: 2.9), at discharge (1.5) and one month after discharge (1.9) showed significant differences between those prior to admission, at discharge, and one month after discharge (p < 0.0001). Hospital admission involved a decrease (p < 0.0001) in the number of patients with polypharmacy criteria (more than four drugs), which persisted one month after discharge (p < 0.01), and in the prescription of polyvitaminic compounds, nonsteroid antiinflammatory drugs, antiaggregants, peripheral vasodilators and antacids (p < 0.03). CONCLUSIONS: A relevant under-reporting of chronic diseases in the discharge report, particularly of those without exacerbations, as well as quantitative (decrease) and qualitative changes in the prescription due to NECD, maintained by the general practitioner one month after discharge. A higher awareness regarding chronic disease is necessary, as well as chronic disease is necessary, as well as establishing communication channels between Primary and Specialized Care. PMID- 9411543 TI - [Effect of methadone or naltrexone on the course of transaminases in parenteral drug users with hepatitis C virus infection]. AB - A prospective study was conducted on the evolution of serum transaminases in 116 patients infected with hepatitis C virus (HCV), parenteral drug abusers, included for 6 months in treatment programs with methadone, naltrexone or "drug free" regimen. Treatment with methadone or naltrexone did not result in a transaminase increase in these patients. In contrast, patients included in the drug-free program appear to have a higher increase in serum transaminase levels than those treated with methadone or naltrexone (146.1 +/- 122.29 vs 91.88 +/- 81.96 and 86.99 +/- 64.26 Ul/ml, respectively). Such an increase originated mainly from patients who anytime during follow-up had a liver necrosis outbreak in the acute hepatitis range, and is offset if, instead of a given transaminase value, we consider a simplified liver necrosis index (1.28 +/- 0.74 vs 0.95 +/- 0.72 and 1.01 +/- 0.65, respectively; p > 0.05). HIV infected patients have serum transaminase values similar to those in HIV-negative patients (80.61 +/- 58.81 vs 113.63 +/- 99.59, respectively; p > 0.05). PMID- 9411544 TI - [Bacteremia in patients undergoing chronic hemodialysis in a 16-year period]. AB - OBJECTIVE: To determine the incidence of bacteremia among patients on hemodialysis, the responsible microorganisms and to describe the predisposing and prognostic factors. METHODS: A retrospective analysis was conducted of 85 episodes of bacteremia occurred from 1979-1994; the episodes involved 71 patients (male/female ratio: 27/44) with a mean age of 58 years (29-80). RESULTS: Eighty seven microorganisms were recovered, which included 61 grampositive cocci (67% Staphylococcus aureus), 25 gramnegative bacilli (52% Escherichia coli) and 1 anaerobe. The mean incidence was 3.1/100 patients on hemodialysis/year (range: 1.1-8.3), higher in patients with interstitial and cystic renal disease. In 52% of cases an intravascular source was detected, associated with vein access for hemodialysis (in 91% there were inflammatory signs at the fistula). In 16 cases (19%) no portal of entry was detected and in the remaining patients the portal of entry had an extravascular origin. Eighty patients received antibiotic therapy and 35 patients required the substitution of the vein access. Thirteen patients died (15%) as a result of bacteremia. The mortality rate was higher among patients developing shock (50%), endocarditis (75%) and in those who had remained for longer than 1,000 days on hemodialysis (45%). Bacteremia accounted for the third known cause of death on dialysis, and was responsible for 11% of deaths occurred during the time of the study. CONCLUSIONS: Bacteremia among hemodialysed patients was mainly associated with Staphylococcus aureus infections at the vascular access. Bacteremia was the direct responsible for 11% of deaths occurred on dialysis. PMID- 9411545 TI - [Long-term course of blood lipids and body mass index in patients with testicular cancer treated with chemotherapy]. AB - OBJECTIVE: To evaluate whether males with testicular cancer treated with chemotherapy including cisplatin (Qt-C) develop an increase in serum cholesterol and triglyceride levels and in the body mass index (BMI), which might pose a cardiovascular risk. PATIENTS AND METHODS: Fifty-six male patients with the previous diagnosis of testicular carcinoma, and apparently cured now, were studied. Thirty-six were Qt-C treated. The median age was 28 years and the median follow-up 81 months. The other 20 patients, with testicular cancer stage I and who did not require Qt-C, did not differ in mean age nor in the median of follow up. Twenty healthy males were also studied, and their cholesterol and triglyceride levels and BMI were compared with those in patients treated with Qt C at the end of follow-up; both group were of similar age. In all patients (prior to diagnosis and yearly up to the end of the study) and in healthy subjects total serum cholesterol and triglyceride levels were measured, as well as BMI (weight/height2). RESULTS: Levels of cholesterol, triglycerides and BMI were not different at diagnosis of testicular cancer, both in patients treated with Qt-C and those not receiving such therapy. When comparing yearly cholesterol and triglyceride levels in both groups of patients during the first 6 years of evolution, no significant differences were observed. Again, no differences were observed between patients treated and not treated with Qt-C at the end of the follow-up period regarding cholesterol (211 +/- 41 vs 219 +/- 44 mg/dl; p = 0.51), triglyceride (128 +/- 57 vs 129 +/- 51 mg/dl; p = 0.94) and BMI (25.7 +/- 3.3 vs 25.5 +/- 3.4 kg/m2; p = 0.86) values. No significant differences were observed in the three parameters between parameters in patients treated with Qt-C at the end of follow-up and in healthy males. CONCLUSIONS: In males with testicular cancer treated with Qt-C, no long term increase in cholesterol, triglyceride, and BMI values was detected, which might predispose to the development of cardiovascular diseases. PMID- 9411546 TI - [Pyogenic hepatic abscess. Review of 59 cases and experience with imipenem]. AB - OBJECTIVES: To study the different etiopathogenic, microbiological, clinical, evolutive, and therapeutic aspects in patients with pyogenic liver abscesses, with a special emphasis in the usefulness of imipenem-cilastatin therapy. MATERIALS AND METHODS: The clinical records of 59 patients with liver abscesses (45 single abscess and 14 multiple abscesses) diagnosed at our institution in the last eleven years were studied. RESULTS: The most common predisposing conditions included biliary (35.6%) and colon (15.3%) diseases, and abdominal trauma (15.3%). The microorganisms responsible for these abscesses included E. coli, Bacteroides spp., and different streptococci. CT and/or abdominal echography were the diagnostic techniques most commonly used. Twenty-three patients were treated with percutaneous drainage and antibiotics, 22 with surgical drainage and antibiotics, 6 with both types of drainage and antibiotics, and 8 exclusively with antibiotics. Twenty-three patients received imipenem (1 g/IV/8 h) and 29 other antibiotics. Twelve patients died and 9 required admission at the ICU. With regard to patients treated with imipenem, 17 (73.9%) cured, 3 of them (one single abscess and two multiple abscesses) without drainage. Two patients treated with imipenem (8.7%) and 4 treated with other antibiotics (13.8%) relapsed. CONCLUSIONS: Imipenem can be a useful antibiotic in association with percutaneous or surgical drainage for the treatment of pyogenic liver abscesses. PMID- 9411547 TI - [Spontaneous peritonitis caused by ascitic fluid with Candida albicans]. AB - Two cases are reported of spontaneous ascitic fluid Candida albicans peritonitis in two patients with liver cirrhosis secondary to HBV. In non of the two cases was there evidence of findings associated with secondary peritonitis, where ascitic fluid Candida infection has usually been reported. PMID- 9411548 TI - [Pericardial tuberculosis complicated with heart tamponade as presentation form of acquired immunodeficiency syndrome]. AB - BACKGROUND: Pericardial tuberculosis is an unusual presentation form of extrapulmonary tuberculosis, even in HIV-1 infected patients. When complicated with cardiac tamponade the prognosis worsens; therefore, early diagnosis and therapy are essential. METHODS: A cross-sectional and descriptive study was carried out of cases with documented diagnosis of tuberculous pericarditis in a cohort of prisoner patients with tuberculosis in the Comunidad Autonoma de Madrid, during a 4-year period (March 1991-March 1995). The recovery and identification of Mycobacterium tuberculosis in our clinic were made on the basis of standard techniques and DNA probes. The clinical and microbiological features of patients with documented diagnosis of tuberculous pericarditis are reported and a bibliographic review on this topic is made (MEDLINE 1985-July 1996). RESULTS: A total of 483 patients were diagnosed of tuberculosis, and 90% were HIV 1 positive. Only four patients, all coinfected with HIV-1, developed tuberculous pericarditis complicated with cardiac tamponade which required drainage and allowed the visualization of acid-fast bacilli and culture of M. tuberculosis in pericardial fluid (in three cases associated with recovery from other specimens). All isolates were initially susceptible to first-line antituberculous drugs. No patient had previously had opportunist infections, although all of them had severe immunosuppression (< 0.200 x 10(9)/l CD4+ lymphocytes). The clinical outcome was favorable with pericardial drainage and the drug regime prescribed; the survival times were 8, 12, 13 and 28 months. The latter patient, on account of inadequate therapy compliance developed multi-resistant tuberculosis. CONCLUSIONS: Tuberculous pericarditis in HIV-1 infected patients usually appears in situations of advanced immunosuppression and usually in the context of disseminated tuberculosis and as a first opportunist infection. Its presentation with cardiac tamponade is unusual and its high morbidity and mortality demand early diagnosis and therapy. PMID- 9411549 TI - [Atypical serologic patterns in hepatitis B virus infection]. PMID- 9411550 TI - [Monocellular myeloproliferative syndromes with histologic protagonist. Primary histiocytosis and mastocytosis as potential monoclonal myelopathies]. PMID- 9411552 TI - [Patient with a history of abdominal trauma presenting digestive intolerance]. PMID- 9411551 TI - [Aortic stenosis in the elderly: treatment strategies]. PMID- 9411553 TI - [Radiculopathy C-8]. PMID- 9411554 TI - [Facial ulcers in a patient with advanced acquired immunodeficiency syndrome]. PMID- 9411555 TI - [Hypodense nodular images in the spleen of a patient with human immunodeficiency virus infection]. PMID- 9411556 TI - [Jod-Basedow phenomenon: who was Dr. Jod?]. PMID- 9411557 TI - [Prevalence of tobacco use in the population and its prevention]. PMID- 9411558 TI - [Chronic neutrophilic leukemia. Blood muramidase]. PMID- 9411559 TI - [Non-traumatic rhabdomyolysis, compartment syndrome, and acute kidney failure caused by heroin]. PMID- 9411560 TI - [Complicated parapulmonary multiloculated effusion cured with intracavitary urokinase after conventional treatment failure]. PMID- 9411561 TI - [Meningococcal infection. A latent entity, but not forgotten]. PMID- 9411562 TI - [Epidemiology and transmission of cysticercosis]. PMID- 9411563 TI - [Neurocysticercosis in a tertiary hospital. New advances in the diagnosis and treatment]. AB - Cerebral cysticercosis is an endemic parasitosis in many developing countries with a great variability in its presentation and prognosis. In the last 37 years (1960-1996) we have studied 18 patients at Fundacion Jimenez Diaz. Sixteen were spanish; ten of them (62.5%) came from rural areas and they all were seen before 1988. Ten patients (56%) were males and seven (44%) females. The mean age was 49 years (range: 22-80). The most common symptoms at diagnosis were: seizures (61%), headache (55%), visual disturbances (39%), symptoms of intracranial hypertension (33%), mental disturbances (33%) and other focal symptoms (33%). CT and NMR showed changes in all cases, and hydrocephalia, presence of brain calcifications and ventricular or subarachnoidal parenchymal cysts were the most common findings. Three patients received antiparasitic therapy (praziquantel and albendazol), two of them associated with surgery. The most common surgical procedure was cyst exeresis and ventriculo-peritoneal shunts. The evolution was favorable in 90% of cases. PMID- 9411564 TI - [Epidemiology of the osteoporotic fracture of the hip in the province of Palencia]. AB - BACKGROUND: Hip fracture is the most severe consequence of osteoporosis. The aim of the present study was to know the incidence of osteoporotic hip fracture in the Palencia province, its direct economical consequences and characteristics associated with the origin episode. PATIENTS AND METHODS: All patients aged over 49 years who had a nontraumatic hip fracture during the second semester of 1994 and the first semester of 1995 were included in the study. An analysis of costs was performed and each patient received a questionnaire to know the circumstances associated with the episode. RESULTS: During the study period the overall incidence of hip fracture was 83/100,000 inhabitants/year, which corresponds to an adjusted incidence of 240.9/100,000 inhabitants older than 49 years (336.8 women and 120.7 men). There was an exponential growth, with peak values starting at 80 years. The female/male ratio was 2.8 and the mean age 80.8 years. Twenty four percent of fractures occurred in institutionalized persons, with an adjusted incidence of 1,107/100,000 inhabitants/year, which corresponds to a relative risk of 13.57 (95% CI: 10.06-18.28). No significant differences were observed between trochanteric and neck fractures. Ninety-seven percent of fractures occurred after a fall, usually in the morning or afternoon (86%), with lateral direction and impact on the greater trochanter (89%). The mortality rate during admission was 5.9%. The mean cost of care during admission was 1,170,000 pesetas. CONCLUSIONS: The incidence of hip fracture in Palencia is slightly higher than the national mean, probably due to populational ageing. The risk of fracture reaches alarming proportions in the institutionalized population. The implementation of efficient preventive measures, particularly among the exposed populations, is necessary. PMID- 9411565 TI - [Spontaneous meningitis caused by gram-negative bacilli]. AB - OBJECTIVE: To analyze the causative factors, clinical and microbiological characteristics, and prognosis in spontaneous meningitis caused by Gram-negative rods in adult patients. METHODS: Descriptive and retrospective study of all clinical records and microbiological findings in patients diagnosed of meningitis by Gram-negative bacilli, from 1973 to 1995, at Fundacion Jimenez Diaz. RESULTS: Twenty patients had spontaneous meningitis caused by Gram-negative bacilli (2.1% of all diagnosed meningitis during this period). Fourteen patients were older than 65 years (range: 36-81; p < 0.05). Ninety-five percent of cases had an extranosocomial origin (p < 0.001). Ninety percent of patients had some underlying disease (p < 0.001). Twelve patients were immunosuppressed. Seven patients had changes in the urinary tract or repeated UTI infections. The most common clinical symptoms were a decrease in consciousness level, fever, and neck stiffness. Cerebrospinal fluid (CSF) in 18 patients showed cellular and biochemical changes. The CSF smear revealed the presence of Gram-negative bacilli in 15 patients. E. coli was the microorganism recovered most frequently. Immunosuppression (p < 0.05), septic shock (p < 0.001) and antimicrobial therapy not including a third generation cephalosporin (p < 0.01) were independently associated with mortality. CONCLUSIONS: Spontaneous meningitis by Gram-negative bacilli is an uncommon infection. It occurs mainly in immunosuppressed patients older than 65 years or with changes in the urinary tract. It usually has an extranosocomial origin. The investigation of CSF is a highly effective for diagnosis. Therapy with third generation cephalosporins has notably improved its prognosis. PMID- 9411567 TI - [Carbohydrate deficient transferrin and other markers of alcohol consumption in the general hospital]. AB - BACKGROUND: The ascertainment of patients who consume important amounts of alcohol admitted to a hospital is essential to prevent medical and psychological complications. Carbohydrate deficient transferrin (CDT) is a new marker of alcohol consumption which requires validation in the hospital setting. METHODS: The values of carbohydrate deficient transferrin (CDT), glutamic oxalacetic transaminase (GOT), gamma glutamil transpeptidase (GGT) and mean corpuscular volume (MCV) were measured in 101 consecutive patients admitted to the Internal Medicine and Surgery Departments. Considering amounts higher than 60 g/day of ethanol for male patients and higher than 40 g/day for female patients as risk consumption, the values for sensitivity, specificity and area under the curve were calculated for the different biological tests. RESULTS: Twenty-six percent of patients reported a consumption of risk. The sensitivity of the tests were lower than 50% and specificities higher than 77%. CDT had the lowest sensitivity (15%) but it was very specific (98%). CDT had a better sensitivity among women than among men. None of the tests had an area under the curve with adequate efficiency levels. CONCLUSIONS: CDT among the hospitalized patients and other biological markers of alcohol consumption have a low efficiency. PMID- 9411566 TI - [Brucellar meningitis as a peculiar form of clear fluid meningitis. Clinical study]. AB - Out of 200 patients diagnosed of acute of chronic brucellosis, cases were first selected who had neurologic involvement (14%) and then those who had brucellar meningitis (CDC criteria) which corresponds to 4% of the total of the series. The clinical course, evolution time, neurologic manifestations and serologic and bacteriologic characteristics in blood and specifically in the cerebrospinal fluid (CSF). This study was contrasted with the literature review and the conclusion is reached that brucellar meningitis is a peculiar form of clear fluid meningitis, not exceptional in our environment considering the prevalence of brucellosis. PMID- 9411568 TI - [Severe Plasmodium falciparum malaria. Description of 5 cases]. AB - In the last few years a considerable number of imported malaria has been reported in Spain, probably due the increased tourism to areas with endemic malaria, particularly with P. falciparum. This is the species more frequently associated with severe complications and the only one capable of causing cerebral malaria. In this report we review five cases of malaria which required intensive care because of their severity. None of the patients had received chemoprophylaxis. In all cases the admission criterion to the intensive care unit was the organic failure of one or more systems (renal failure and disseminated intravascular coagulation [DIC] mainly) or the presence of changes in the central nervous system. Parasitemia at admission was higher than 5% in all patients. One patient died on account of cerebral malaria. Only one patient had severe complications not directly associated with malaria. In patients who already have severity criteria, a negative parasitemia test during the clinical course does not necessarily implies a clinical improvement nor does it exclude the emergence of complications. On the other hand, a low parasitemic degree is never a contraindication for admission to the intensive care unit when severity criteria are present. PMID- 9411569 TI - [Influence of the quantity and type of carbohydrates consumed in the regulation of body weight]. PMID- 9411570 TI - [Autoimmune chronic thyroiditis]. PMID- 9411571 TI - [A young woman with a mass of soft tissues in the posteroinferior region of the leg]. PMID- 9411572 TI - [A young woman with headache worsening on the standing position and specific findings in magnetic resonance]. PMID- 9411573 TI - [Necrotic lesions in the fingers in an HIV positive patient]. PMID- 9411574 TI - [Persistent tuberous plaque in a finger]. PMID- 9411575 TI - [Immature teratoma grade 1, stage 1C in left ovary, with mature cystic teratoma in right ovary]. PMID- 9411576 TI - [Acute lupus encephalitis associated with serum anti-NUMA]. PMID- 9411577 TI - [Infrequent forms of meningococcal infections]. PMID- 9411578 TI - [Apolipoprotein E and Alzheimer disease (reply)]. PMID- 9411579 TI - [Patients' and relatives' opinion and knowledge about electroconvulsive therapy: implications for nursing]. AB - The aim of this report was to verify in-patients and their relative's opinion and knowledge of the use of electroconvulsive therapy. By means of a quantitative analysis, the use of this treatment was shown to be accepted despite the individuals knew little about it. It was observed that the knowledge of this treatment had influence over individual's opinion about it. Thus nurses play an important role on the instruction of these individuals regarding this treatment. PMID- 9411580 TI - [Use of the Glasgow Coma Scale and the Jouvet Coma Scale to evaluate the level of consciousness]. AB - The Glasgow Coma Scale (GCS) and the Jouvet Coma Scale (JCS) have been evolved for assessing the depth and duration of impaired consciousness and coma. The analysis and the utilization of these scales have showed that they are complementary. The GCS is more sensitive when there is a more intense loss of consciousness, whereas the JCS shows its sensitivity better in the states close to normal. This study was aimed to compare the results obtained from the evaluation of the consciousness level by the utilization of the two scales. The comparison was done within a prospective study with 48 patients, all of them over 18 years old, interned in three intensive care units of different hospitals in the city of Sao Paulo. The evaluations were done daily by the researchers and the scales applied in sequence totaling 5 minutes. Each scale was applied in 106 evaluations, and the results showed a statistically meaningful difference between the GCS and the JCS as to the indication of alteration in the consciousness levels. In 37.74% of the evaluations done with the JCS there was an indication of alteration in the consciousness level, whereas with the GCS the alteration was present in only 23.58% of the evaluations. Another important observation about the utilization of both scales was that people whose scores were between 9 and 10 in the GCS had had an stronger indication of alteration of consciousness level by the same scale, while those with scores between 12 and 15 had a stronger indication of alteration in the consciousness level by JCS. When using GCS there has been the application of the non-testable (NT) in 20% of the evaluations. This did not occur when using the JCS. However it is believed that specific conditions of that particular group might have led to that result as well as specific characteristics of groups of patients might favor the utilization of different scales to evaluate the consciousness level. Therefore the final choice between such scales should consider the conditions and the peculiar characteristics of the clientele to be evaluated and not individual or health department services preferences. PMID- 9411581 TI - [Quality of life in the elderly: Study in a public recreation center for aged persons]. AB - The purpose of this study was to analyze quality of life of the users of a recreation institution for old age people. The results led to the conclusion that the majority of the intervieweds held a good quality of life, according to the subjective and objective indicators analyzed in this study. PMID- 9411582 TI - [White coat hypertension under outpatient conditions: indications for ambulatory monitoring of arterial pressure]. AB - Ambulatory blood pressure monitoring (ABPM) is to be considered an excellent diagnostic tool, progressively used by the clinicians in their routine medical practice, in order to assess both medical diagnosis and antihypertensive treatment efficacy. ABPM permits to distinguish with a high level of accuracy the true hypertensives, from the false hypertensives or from subjects with isolated clinical hypertension, the old so-called "white coat hypertension". Prognostic implications of hypertension and target organ involvement can be assessed more accurately with ABPM than with basal blood pressure measurement. ABPM also permits an excellent approach to 24-hour blood pressure load, establishing different categories of circadian curves. In the same way, the 24-hour monitoring observation period during a normal daily life, helps to identify the hot moments of circadian period, correlating them, with changes in blood pressure, and subsequently with cardiovascular events, particularly in the early morning hours, during the so-called circadian vulnerable period, in which a significant increase in both cardiovascular events and stroke are usually observed. Nevertheless, some limitations are still linked to ABPM systems and equipments; on the one hand, the lack of a cut-off point, to establish the limits of true normally, and on the other one; the today intermittent character of ABPM measurements, do not permit an exact assessment of 24-hour blood pressure profile, containing all the hemodynamic changes. Several ongoing studies, will bring in the near future the very necessary answers to all these interrogants. PMID- 9411583 TI - [Conditions leading patients to the hypertensive cardiac units]. AB - The heart, as a "target" organ of Hypertension, acquires a "central" role in our daily practice from different angles of view: diagnosis, classification, therapy and prognosis of Hypertensive Cardiopathy. A computerized database provided by the Working Group of Hypertension of the Spanish Cardiac Society should unify the actions of recently established Units of Hypertensive Cardiopathy. Those patients, who are selected in the Primary Health Center, referred to the Cardiology Service would be theoretically allocated in one of four headings: lef ventricular hypertrophy, left ventricular disfunction, ischemic coronary disease and electric instability. The fundamental bases standing current management of this unique and colorful cardiovascular pathology are detailed. PMID- 9411584 TI - [Arterial hypertension and exercise]. AB - Both dynamic and static exercise increase blood pressure in normotensive and hypertensive patients, but the change varies among these two main forms of exercise. The validity of the blood pressure response to acute exercise as a predictor for the future development of hypertension in normotensives, or of the degree of target organ damage in hypertensives is still not clear. This acute response could be an independent predictor of mortality or cardiovascular events in the normotensives. The postexercise hypotension could work as a beneficial mechanism to reduce the reduce the rise in pressure that occurs with time. While epidemiological studies suggest an inverse relationship between physical activity or fitness and blood pressure, longitudinal studies seem to confirm the hypotensive effect of dynamic aerobic training. Though possibly in a lesser degree, strength training has also been proved to be effective. Among the responsible mechanisms, a decrease in sympathetic nerve activity is most likely involved. Hypertension seems to be also associated with a lower maximal capacity, that could be even more adversely affected by the action of certain antihypertensive drugs such as diuretics or beta-blockers. All of these aspects have led to the acceptance of physical exercise among the non-pharmacological measures for the treatment of hypertension. PMID- 9411585 TI - [Resources for the diagnosis of hypertensive cardiopathy at the primary care level]. AB - Hypertensive cardiomyopathy is nowadays the most precious, prevalent and fatal condition of all cerebral, renal and arterial complications that leads to arterial hypertension. Left ventricular hypertrophy is the basis of the macroscopic structural damage that belongs to this entity. Basically, this complication produces, in the daily practice, alterations in the systolic and dyastolic functions, myocardial ischemia and arrhythmias. At present, it is obvious that we need to take better advantage of resources to diagnose hypertensive cardiopathy, and that the cost of explorations are lower must imply an agreement between cardiologists and general practitioners. In this article, we review the resources available at general practice level for the efficient diagnosis of the complications produced by hypertensive cardiopathy. PMID- 9411586 TI - [Arterial hypertension and diabetes]. AB - Abnormalities of glucose, insulin, and lipoprotein metabolism are common in patients with hypertension. This constellation of risk factors may be recognized at young ages and is at least in part heritable. The insulin resistance and the compensatory hyperinsulinemia could be primary events, and enhanced sympathetic activity and diminished adrenal medullary activity would be important links between the defect in insulin action and the development of hypertension and the associated metabolic abnormalities. But not all hypertensive patients have insulin resistance. It is possible that insulin resistance, and compensatory hyperinsulinemia have major roles in the regulation of blood pressure in susceptible subjects predisposed to hypertension by heredity or environmental factors. Considerable evidence, both in experimental animal models and in humans, points to hypertension as of critical importance in the pathogenesis of severe diabetic heart disease. In diabetic hypertensive cardiomyopathy, coronary artery disease as well as structural and functional abnormalities are more pronounced than would be expected from either process alone. The hypertension increases the risk of diabetic nephropathy in non-insulin-dependent diabetic patients. The microalbuminuria is a powerful predictor of mortality in these patients. It seems that angiotensin-converting-inhibitors have efficacy in postponing nephropathy in hypertensive non-insulin-dependent diabetic patients. In patients with hypertension and diabetes, additional clinical trials are required to identify those interventions that will most effectively reduce not only overall risk but also definitive cardiovascular disease endpoints. PMID- 9411587 TI - [Cardio-protection and cardio-reparation in systemic arterial hypertension]. AB - Left ventricular hypertrophy in systemic hypertension is an important independent risk factor for adverse cardiovascular events. Hypertrophy is due to increase of the myocytic and non-myocytic compartments of the myocardium. The changes in architecture, shape and function of the left ventricular is considered as remodelling. The concept of cardioreparation implies a restoration of structural (regression of both intersticial and perivascular fibrosis and myocyte hypertrophy) and functional consequences of remodelling. The concept of cardio protection implies prevention of remodelling. Remodelling is caused not only because high pressure but there are many other factors, mainly hormonal, as activation of renin angiotensin aldosterone system promoting nuclear oncogens. Functional changes are mainly due to fibrosis with diastolic impairment at a first step and dilatation later. Therapeutic goal in systemic hypertension is to achieved, not only a normal pressure but a normal structural and functional heart. PMID- 9411588 TI - [Cardioprotective and cardio-reparative drugs]. AB - The left ventricular hypertrophy is a deleterious consequence of the arterial hypertension, recognized as independent risk factor. The left ventricular hypertrophy has a myocytic component, of hemodynamic origin, where all the antihypertensive treatments known would be active, in greater or smaller degree, and it's been demonstrated the benefit of the intervention in the sense of the fact that exist improvement indications of the morbi-mortality (cardio protection). The left ventricular hypertrophy has at the same time a fibrotic component, of neuro-hormonal origin that can be reversed (cardio-reparation) fundamentally by influence of the ACEIs, that would improve furthermore all the components of the called "hypertensive syndrome". PMID- 9411589 TI - [How to choose and adequate antihypertensive drug depending on the type of the existing heart damage?]. AB - The main goal of the treatment for hypertensive vascular disease is to reduce the morbidity and mortality that follow the disease. In the patient with heart disease, the choice of antihypertensive treatment will depend on several factors, all of which must be considered prior to it: type of cardiopathy and complications, pharmacokinetics of the drug-selected and its side effects, interactions with specific treatment for the main heart disease, positive or negative interactions with risk factors and, finally, its prognostic benefits. In the present study we briefly analyze this considerations in relation to different diseases such as ischemic heart disease, ventricular dysfunction (hypertrophy, systolic and diastolic dysfunction), heart rhythm disorders (sinus node dysfunction, supraventricular and ventricular ectopies), vascular pathology (cerebral and peripheral vasculopathy) and risk factors (diabetes, dyslipemia, obesity). Based on this considerations, several recommendations are done in order to choose the best antihypertensive drug in such cardiovascular diseases. PMID- 9411590 TI - [QKD: a new parameter for clinical research on hypertension]. AB - Structural and functional heart changes induced by hypertension are easily detected nowadays by ecocardiography-Doppler. Nevertheless, this diagnostic tool is less conclusive to evaluate the large arteries modifications during hypertension. High blood pressure induces from the beginning architectural changes in all artery beds, characterized by an increase in wall thickness, reversing secondary the ratio wall thickness/lumen radio. The increase in wall stiffness as a consequence of cellular hypertrophy and hyperplasia, modifies substantially both, distensibility and compliance, two major factors in determining the grade of left ventricular hypertrophy and performance. Evaluation of pulse wave velocity implemented with a new algorhythm (QKD) permits by non invasive ambulatory long period of time method, an indirect approach to the arterial functional state in hypertension and aging, establishing in some instances, prognosis criteria, with an accuracy even better than that given by the 24-hour load pressure index. PMID- 9411591 TI - [Hypertensive cardiopathy and arrhythmias]. AB - The anatomical and tissue changes caused by hypertension are responsible of the higher incidence of atrial and ventricular arrhythmias as compared to normal population. Hypertension, when associated with atrial fibrillation, becomes an important risk factor for systemic embolism. Atrial dilatation and/or fibrosis because of haemodynamic overload due to left ventricular hypertrophy are the arrhythmic substrate. The higher incidence of ventricular arrhythmias is related to left ventricular hypertrophy. Ventricular premature beats, frequent and/or polymorphic in most of the cases, and short runs are the usual picture. Sustained ventricular tachycardia is seldom documented. The substrate is created by hypertrophy itself, resulting in conduction disturbances favoring reentry. Associated myocardial ischemia plays an important role in the genesis of ventricular arrhythmias. Left ventricular hypertrophy is associated with an increased incidence of total cardiac death and sudden death. Ventricular arrhythmias are suggested to be a poor prognostic factor. The aim of this article is to offer a simplified review of the meaning of cardiac arrhythmias in the hypertensive patient, and to give some clues to the practical approach in the clinical setting. PMID- 9411592 TI - [Regression of left ventricular hypertrophy in hypertensive patients]. AB - Left ventricular hypertrophy associated with systemic hypertension differs from left ventricular hypertrophy initiated by other pressure overload diseases. Its development depends not only of hemodynamics aspects but of biochemical factors. Many studies have demonstrated a close link between left ventricular hypertrophy and cardiovascular morbidity and mortality. For that reason the idea of reversal of left ventricular hypertrophy has been a goal of the antihypertensive treatment. From the literature review has been established that the most classes of antihypertensive medications reduce the left ventricular mass, though there is a variation in required duration of treatment. At this point the angiotensin converting enzyme inhibitors, probably because a double effect: hypotensive and blockers of the trophic stimulus of angiotensin II, seemed to be the most potent for reducing the left ventricular mass. Still we don't know if reversal of left ventricular hypertrophy, by the antihypertensive treatment, reduce independently the cardiovascular risk. PMID- 9411593 TI - [Leukotriene antagonists: a new approach in the treatment of asthma]. AB - Inflammation plays an essential role in the genesis of airflow obstruction and bronchial hyper-reactivity in the early stages of clinical asthma. The treatment of bronchial inflammation has become an essential element in the therapeutic strategy and principally rests on inhaled glucocorticoids. Amongst a number of inflammatory mediators leukotrienes occupy a privileged place by the power of their inflammatory and constrictor effects on bronchial smooth muscles. These properties have justified the clinical development of inhibitors of their synthesis and of specific antagonists to their receptors. Leukotriene antagonists are specific for a sub type of leukotriene receptors C4, D4 and E4 which is implicated in the majority of the bronchial constrictor and inflammatory effects of leukotrienes. The antagonists of Cys-LT1 receptor but also the inhibitors of the leukotriene synthesis exert an additive bronchodilator effect to those of B2 stimulants confirming an efficacious protection vis a vis bronchial provocation tests and above all they improve the clinical scores, lung function and also enable a decrease in the consumption of beta 2 agonists. The marketing of these products represents a major event because it corresponds to the advent of a new therapeutic class. The ease of administration by the oral route, their demonstrated efficacy and their good tolerance profile (in particular for ICI 204.219, and antagonists to Cys-LT1 receptors) are elements which foresee a success for this new asthmatic treatment. However numerous studies, notably comparative studies vis a vis reference treatments will be necessary to define their place in the strategic approach to the treatment of asthma. PMID- 9411594 TI - [Allergies: determinants of T2 lymphocyte polarization and desensitization mechanisms]. AB - Distinguishing two populations amongst T lymphocytes, namely Th1 and Th2, has enabled a better understanding of the pathophysiology of inflammatory diseases most notably immunoallergic disorders. These two populations, initially described in mice, are defined by the profile of cytokines which they produce. Th1 lymphocytes secrete amongst others IL-2 and IFN-gamma and the action exercised by these cytokines are involved in cellular reactions. Th2 lymphocytes are involved in humoral responses and immediate hypersensitivity secreting IL-4 and IL-5. In man the distinction between the two populations is more subtle than in the mouse, in particular for the same cell there is a possibility to pass from one type of cytokine to another. Thus the importance of factors which influence the orientation towards Th1 or Th2 type amongst the T lymphocytes. What are these factors? Can one use these to change the response to achieve a therapeutic goal? These are the questions that we propose to touch on in this analysis broaching the induction of the Th2 response. Certain responses are linked to the individual and concern their heredity. Others are the cells themselves which are involved in antigen presentation, the presenting cell and the HLA molecules, peptide antigen and T cell receptor. Again there are others which influence the presentation without being directly involved: these are the cytokines already present in the biological milieu and engaged co-receptors. All these factors act in concert, perhaps with others which are not yet known and account for the complexity of the response. Desensitisation is a treatment whose efficacy is recognised in certain precise indications. During the course of treatment, there is a diminution of IgE to the benefit of IgG and a diminution of the eosinophil count induced by specific provocation. These effects may be related to a decrease in the production of IL-4 and IL-5 respectively, which has been found in several recent studies. Desensitisation also shows that in spite of its complexity, one can reorientate towards a Th1 response based on spontaneous Th2 response. In this respect desensitisation constitutes a human model for the study of orientation of the Th2 immune response. In the future better understanding of the mechanisms of the allergic reaction will enable new treatments to be developed based on the use of antigenic peptides or using cytokines and inhibitors of cytokines. PMID- 9411595 TI - [Skeletal muscle abnormalities in chronic obstructive lung disease with respiratory insufficiency. Value of P31 magnetic resonance spectroscopy]. AB - 31P magnetic resonance spectroscopy (31P MRS) is a non-invasive method to evaluate high energy compounds [adenosine triphosphate (ATP), phosphocreatin (PCr), inorganic phosphates (Pi)] and intracellular pH (pHi) of skeletal muscle during exercise and recovery. It is a clinically applicable method of investigation for severe COPD patients with respiratory failure since exercise is limited to a single group of muscle (calf). Pronounced alterations of muscular metabolism have been shown in these patients: (1) reduced aerobic capacity (as reflected by the ratio of Pi/PCr as a function of power and changes in recovery kinetics of PCr after exercise and (2) increased anaerobic metabolism (reflected by a decrease in intracellular pH). Four different studies reveal similar abnormalities. Acute oxygen administration partially improves these parameters, suggesting that other factors in addition to hypoxaemia may contribute to the metabolic impairment. The effect of increased physical activity on these abnormalities deserve further investigations. PMID- 9411596 TI - [Influence of the inhaler device on bronchodilatation in the asthmatic child]. AB - The use of spacer devices is recommended in asthmatic children for inhaled therapeutics. Therefore, in vitro studies prove the dependent-device delivery of the drug. The aim of this study was to compare, in vivo, the effect of 200 micrograms of albuterol, delivered via one of the five spacer devices currently marketed in France (Aerochamber, Aeroscopic, Babyhaler with a face mask, Nebuhaler or Volumatic) and assessed by the induced peak expiratory flow (PEF) change. One hundred asthmatic children were recruited and randomized in 5 groups. The mean age was 8.9 +/- 3.3 years. Each group was comparable regardless of gender, height, weight, characteristics of asthma and baseline PEF. The maximal change in PEF was obtained with Babyhaler (14.9 +/- 12.8%; p = 0.009). The increase in PEF elicited with Aeroscopic was 9.7 +/- 10.2%. The others spacer devices did not offer a change greater than the variation of PEF in the studied population. Further studies, concerned with a measurement of drug deposition or with an assessment of its use in obstructive episodes of asthma, are required, but Babyhaler with a face mask, usually reserved to infants, deserves to be advised to older children for salbutamol intake. PMID- 9411597 TI - [Role of nocturnal oximetry in screening for sleep apnea syndrome in pulmonary medicine. Study of 329 patients]. AB - Nocturnal oximetry can show nocturnal oxygen desaturation. This examination was proposed as an investigation for the early detection of the sleep apnoea syndrome (SAS). We have compared the results of nocturnal oximetry and polysomnography in 329 consecutive patients seen in the department of thoracic medicine for the early detection of the SAS between June 1990 and June 1995. The diagnosis of SAS was confirmed at the time of polysomnography using an hypopnoea/apnoea index (IAH) greater or equal to 15 per hour. Two parameters of oximetry were well correlated with IAH less than 15 per hour: if the mean oxygen saturation is greater than 92% and for less than five per cent of the time of the examination there was a saturation of less than 90%. The sensitivity was 89.7% and the specificity was 57.8%. Among the 48 false positive cases on oximetry 17 patients were found to be suffering from COPD and 31 patients were probably suffering from a syndrome of upper airways resistance or possibly from the hypoventilation obesity syndrome. Amongst the 22 false, negatives to oximetry 10 non COPD patients with an IAH of greater than 30 per hour and diurnal somnolence had important anomalies of the oro-pharyngeal pathway as the origin of their nocturnal apnoea. The 12 other false negatives were patients with moderate SAS with an IAH of between 15 and 20 per hour. Logistical analysis has shown the association of the two oximetric criteria (mean oxygen saturation or percentage of time with a saturation of less than 5%) with clinical criteria (body mass index and formation on diurnal somnolence from a questionnaire) would enable a probable diagnosis of SAS in 75% of cases. Our study shows that nocturnal oximetry used an early diagnosis test, associated with clinical and respiratory function data enables the number of requests for polysomnography to be reduced. PMID- 9411598 TI - [Educational diagnosis in asthma. Descriptive results of a questionnaire survey]. AB - The object of this work was to carry out an audit on an asthmatic's knowledge of their disease and of the risk factors, their usual therapy and of therapy for acute exacerbations as well as their need for information and education. The study was carried out by a survey of 327 adult asthmatics who were in consultation with specialist physicians in thoracic medicine. The information was gathered using an anonymous questionnaire completed by a nurse. The doctors looking after these patients had given their opinion on the severity of their asthma and the educational needs of their patients. The population studied had an average of 47 +/- 17 years. Forty seven per cent of the subjects were masculine. The general level of education was high. Seventy five per cent of the patients were from an urban environment. The duration of the asthma was on an average of 19 years. Half of these patients had already been in hospital for a serious attack. One third of the patients were considered by their doctors as having severe asthma. The patients were good at distinguishing the level of severity of their acute exacerbations and adapted their therapy on their own initiative in appropriate fashion based on the degree of severity of their disease. The enquiry revealed that patients take some liberties around the basic treatment that they had been prescribed. It also showed some defects or errors in their knowledge of the disease and the appropriate approach vis-a-vis the disease. One noted for example that one third of the asthmatics had already stopped the treatment before the date fixed by their doctor. In a general, patients who were surveyed were interested in their health problems with their health and hoped to be better informed both on asthma and its treatment. This study was biased towards an interest in the understanding of the education in order to manage the asthmatic population better. PMID- 9411599 TI - [Association of idiopathic eosinophilic pleurisy and a severe pericardial effusion]. AB - A 52-year-old man presented with a two month history of left sided chest pains related to a pleurisy which showed an 18 per cent eosinophilia on pleural aspiration. He was hospitalised one month later and at that stage was very dyspnoeic and there was a bilateral pleural effusion associated with a significant pericardial effusion causing compression. A pleuro-pericardial drainage eased the patient's symptoms. The pleural fluid showed an 87 per cent lymphocytosis and a complete diagnostic work up was negative thus a diagnosis of idiopathic eosinophilic pleurisy with an associated pericarditis was made. Steroid therapy was instituted using half a milligram per kilo and this led to a rapid functional improvement and no recurrence of the effusions were noted. The treatment was progressively stepped down over the next five months and the patient's condition was satisfactory without any recurrence at nine months. PMID- 9411600 TI - [An unusual cause of aorto-bronchial fistula: tuberculosis aortitis]. AB - The aortic rupture in the pulmonary parenchyma or the bronchi rarely results in an haemoptysis. It means in most of the cases the rupture of an aortica aneurysm. We relate the observation of a aorto-bronchial fistula from a tuberculosa origin in an old woman case. Although the tuberculosa aortitis is becoming very exceptional, it still remains the cause of aorta rupture, with the formation of a false aneurysm which is rapidly fatal and so, it is important to search for it before any capricious haemoptysis. PMID- 9411601 TI - [Benign clear cell tumor ("sugar tumor"). An unusual cause of intrapulmonary coin lesion]. AB - The authors report a case of a 25 years old woman in whom a coin lesion was fortuitously discovered. Initial investigations were negative and an exploratory thoracotomy was performed which enabled a benign clear cell tumour of the lung to be found (sugar tumour). This rare benign tumour whose cellular origin remains indeterminate is in general discovered in a fortuitous manner after a chest x-ray has been performed showing a round peripheral opacity. The diagnosis is confirmed following the excision of the tumour, complementary examinations are not helpful. PMID- 9411602 TI - [Pericardial effusion and mediastinal "tumor": case report of an unusual association]. AB - A young man aged 26 was admitted as an emergency to the thoracic surgical unit for a pericardial tamponade associated with a tumour in the left anterosuperior mediastinum. After pericardial drainage by the sub-xiphoid route a left anterior mediastinotomy showed that the tumour had disappeared and a flabby diverticulum of the pericardium was found which was resected. Pericardial diverticulae are coelomic cysts which are usually considered to be congenital in origin similar to the pleuro-pericardiac cysts to which they are attached. The clinical history, which had a particularly dramatic and alarming onset is in favour of an acquired disease which is equally possible in this type of lesion. PMID- 9411603 TI - [Unusual radiological symmetry]. PMID- 9411604 TI - [Unusual radiological diagnosis of pleurisy]. PMID- 9411605 TI - [Central venous catheters with totally implanted injection ports: use and surveillance]. PMID- 9411606 TI - [Bibliometry of biomedical periodicals]. PMID- 9411607 TI - [Fiberoptic bronchoscopy and diagnosis of lung diseases in intensive care]. PMID- 9411608 TI - [Lung volume reduction surgery in emphysema]. AB - Lung volume reduction surgery in emphysema has, as an objective, the reduction of dyspnoea and an increase in the exercise tolerance in patients with respiratory insufficiency suffering from diffuse emphysema. In principle the resection of the most diseased areas of emphysema leads to improvement in the mechanical properties of the emphysematous lung and correct pulmonary hyperinflation. The respiratory function benefits both objective and subjective, produced by surgery are real but transitory and inconstant depending in particular on the evolutionary profile of the emphysematous disease. The indications should be further refined and an objective comparison of different surgical techniques has not been achieved. The impact on the quality of life for these patients is unknown. PMID- 9411609 TI - [Inhalation route for administration of systemic drugs]. AB - With aerosolized administration of medicines, direct access to the lung is possible. Depending on the intrapulmonary behavior of each molecule, the aerosol mode of administration makes possible either a pulmonary topical action or a systemic action, thanks to the lung's wide vascular network. An inhaled molecule's pulmonary behavior is not predictable and must be studied on its own. Some medicines with a systemic aim have a topical effect after aerosol administration: furosemide, amiloride, morphine, interferons, cyclosporin and prostacyclin. Other medicines have systemic effects after aerosol administration: insulin, growth hormone, LHRH, calcitonin and nicotine. Heparin can be classified in either group depending on the dose and type of use. PMID- 9411610 TI - [Dose-effect of chemotherapy in bronchial small-cell cancer]. AB - Treatment of patients with small-cell lung cancer (SCLC) remains disappointing despite high initial complete response rate. The dramatic initial chemosensitivity of tumor cells is frustrated by the early emergence of drug resistant clonogenic cells, regardless of front line treatments. Enhancement of dose-intensity (DI) can be achieved by various strategies: initial or late intensive chemotherapy with the use of hematological supports, moderate increase in the initial dose of chemotherapy or intensive weekly chemotherapy. Although the dose relationship is fairly well established regarding the response rate, the effect of DI on survival is not clearly shown in most of the different trials. But the effect of a moderate increase in the initial dose of cisplatinum and cyclophosphamide on survival, opens new directions in the therapeutic strategy of SCLC. The contribution of hematopoietic growth factors and reinfusion of hematopoietic progenitors may be of great interest in the management of this disease. PMID- 9411611 TI - ["Primary" diffuse interstitial fibrosis in coal miners: a new entity? Study Group on Interstitial Pathology of the Society of Thoracic Pathology of the North]. AB - It is well known that silica exposure leads in an experimental model to the development of an acute fibrotic process. In human beings two main observations have already been done: (1) silica exposure is frequently associated with the development of connective tissue disease (CTD), especially progressive systemic sclerosis; (2) 10 to 20% patients with CTD developed pulmonary fibrosis. In this context we report 26 cases of coal miners who presented with clinical, radiological, biological and functional characteristics mimicking idiopathic pulmonary fibrosis (IPF), with or without associated coal worker's pneumoconiosis (CWP). All were men; mean age was 68 +/- 9.2 years. Twenty-three were smokers. Duration of exposure was 28.8 +/- 9.1 years. All the patients had dyspnea (stage III, IV in the NHYA classification) and diffuse crackles. Eleven out of 26 had finger clubbing. Computed tomography showed honeycombing (23 cases), and/or ground glass opacities (6 cases) with bronchiectasis (3 cases) predominant in the lower lobes; 19 had radiological signs of CWP, micronodules (n = 16) and nodules (n = 3) predominant in the upper lobes. BAL exhibited an increased % of neutrophils (11.9 +/- 16.1%). Lung function demonstrated a restrictive pattern (TLC = 73 +/- 15.6% and VC = 80 +/- 18% of predicted values) associated with a decreased DLCO (51.8 +/- 23.6% of predicted values) and hypoxemia (at rest = 66.5 +/- 11.2 mmHg, upon effort = 56 +/- 12 mmHg). Lung biopsies were performed in four cases and demonstrated interstitial fibrosis of intraalveolar septum with an accumulation of immune and inflammatory cells similar to the one described in IPF. The association between IPF and silica exposure with or without associated CWP points out the problem of legal recognition of idiopathic-like pulmonary fibrosis as a complication of the occupational exposure of coal workers. PMID- 9411612 TI - [Thoracoscopic approach in pulmonary nodules: a prospective evaluation of a series of 120 patients]. AB - More and more pulmonary nodules are currently approached via thoracoscopy. We have evaluated the results and the morbidity of a consecutive series of 120 patients operated on by a single surgeon. PATIENTS AND METHODS: Hundred twenty two nodules have been approached thoracoscopically in 120 patients. The average size of these nodules was 16 mm (3-30 mm). A pre-operative localisation technique has been used in 61 patients (50%). The procedures were as follows: biopsy (6 cases), wedge-resection (110 cases). A video-assisted lobectomy has been performed in 26 cases. RESULTS: The mortality rate was 0.08% (One case of ARDS in the post-operative course of a video-assisted lobectomy). Intra-operative morbidity rate was 1.6% (2 cases of haemorrhage requiring a thoracotomy and the post-operative morbidity rate was 5%. Six procedures were converted to thoracotomy 55%). The nodules have been localised in all cases but 2 (1.6%). The mean post-operative stay was 4.6 days in the whole series and 3.2 days in the series of patients with a simple biopsy or wedge-resection. COMMENT: The morbidity rate of thoracoscopic resection of lung nodules is very low and decreases with surgeon's experience. Experience allows one not to use a localisation technique in many cases, but the later remains helpful in small size nodules. It allows for a safe, rapid and accurate procedure to be performed. The need for a mini-thoracotomy is very rare. Mastering the techniques of radiological localisation techniques, thoracoscopic biopsy and wedge resection as wall as video-assisted lobectomies should make it possible for thoracoscopic resection of lung nodules to fulfil the criteria of a minimally invasive operation. PMID- 9411613 TI - [Asthma in private pneumology practice]. AB - We assessed asthma severity in patients attending private practice chest specialists, studied the factors related to classification by physicians, and described medications prescribed. 545 chest specialists scattered throughout France, included the patients examined from 3 to 28 May 1993 (N = 14,865). Besides a classification of asthma severity in four classes (mild, moderate, moderately severe, and severe), questionnaires included 20 questions on the history and characteristics of asthma, lung function level and medications prescribed. The proportion of mild asthmatics was 55% among the 3,620 children (aged 6 to 15 years), 42% among the 6,479 young adults (aged 16 to 45 years), and 18% among the 4,766 older adults (aged 46 to 75 years). Followed-up patients were considered more severe than new patients among adults, but not among children. The factors related to asthma severity were impaired FEV1, history of hospitalization, critical care and emergency visits for asthma, limitation of physical activities, and, to a latter extent, symptoms between exacerbations, frequent asthma attacks and daily use of beta 2-agonists. Anti-inflammatory drugs were prescribed to practically all patients from grade 2 (moderate): steroids increased whereas sodium cromoglycate and nedocromil decreased with increasing severity. This study provides a valuable estimate of the classification and medications prescribed to asthmatic patients examined by 50% of private practice chest specialists in France. PMID- 9411614 TI - [Program of Medical Information System and diseases of the respiratory tract (CMD, No.4). Study of 1,020 hospital stays out of the total of 18,253 hospitalizations at the Centre Hospitalier de Saint-Gaudens in the framework of the National Study of Costs]. AB - OBJECTIVE: To analyse hospital activity, the characteristics and course of patients having been classified, by the programme of medical information system (French PMSI) in the major diagnostic category of respiratory disorders (CMD n(o) 4). STUDY: Using the files created by Saint-Gaudens Hospital (145 acute beds, 7000 annual admissions) for three years within the context of the national study of the cost of medical activity. A descriptive statistical analysis then an explanation of the variations of consumptions by stay and by patient. RESULTS: In CMD n(o) 4 there were 26 homogeneous groups of patients (GHM) representing 1020 hospital admissions (out of a total of 18,253) of whom 76 were admitted as an emergency, 63 were in the thoracic medicine group and 27 had a spell in intensive care, 73% were discharged home and 9.8% died. CMD n(o) 4 represented 9% of the hospitals synthetic activity index (ISA). Of these 822 patients had a mean age of 70, 60% of them had chronic disease and 20% were living alone. The variations of cost had been explained by GHM (P < 0.001), test of variants [ANOVA]) as well as by complementary indicators (performance status WHO) and a simplified index of severity. Men, elderly subjects and those with chronic disease consumed more. Hospital activity determined from CMD n(o) 4 has been compared to an extraction of stays based on pure diagnostic medicine (thoracic medicine) of the principal disorder: important differences were apparent, explained in large part by the method of admission, by the PMSI, respiratory tumours and ambulatory care. CONCLUSION: An analysis of activity in thoracic medicine has been able to give an over view of the hospital with the reserves linked to the system which were essentially economic and should be able to be compensated by later improvements seeking to give a more medical aspect to the description of hospital stays. PMID- 9411615 TI - [Chylothorax and sarcoidosis]. AB - The association of sarcoidosis and chylothorax is rare. A patient aged 64 presented with mediastinal sarcoidosis. Two years after the diagnosis he developed a right chylothorax. The computed tomographic examination showed a retraction of the right hemithorax associated with large mediastinal nodes, without signs of interstitial lung disease. Pulmonary function tests revealed a restrictive ventilatory defect with a Forced Vital Capacity (FVC) of 53% and a Forced Expired Volume in one second (FEV1) of 54% from the predicted values; the FEV1/FVC ratio was 78%. Steroid therapy was started at 1 mg per kilogram per day for three months then progressively decreased to 15 mg per day. Inspite of the steroid therapy there was as well biological evidence of continued disease activity (bronchoalveolar lavage lymphocytosis and increased serum angiotensin converting enzyme), as persistence of pleural effusion and mediastinal adenopathy; the contraction of the right hemithorax increased. Restrictive ventilatory defect worsened with a FVC of 44% and an FEV1 of 47% of the predicted values. The presence of a significant contraction of the hemithorax and of a severe restrictive ventilatory defect suggested the existence both of pleural fibrosis and of a compression of the main lymphatic pathways responsible for the chylothorax. PMID- 9411616 TI - [Occupational asthma caused by buckwheat flour]. AB - Buckwheat flour, mainly used for pancakes, may induce asthma following inhalation and anaphylactic reactions following ingestion. These allergic reactions are mediated by specific IgE and may be confirmed by skin test and radio-allergo sorbent test. The occupational asthma of a patient working in pancake restaurant was confirmed by specific challenge test with a computerised device to generate particles. A very small amount of buckwheat flour (10 micrograms) induced an immediate fall of the FEV1 to 56% of the initial value. No bronchial reaction was observed with lactose nor with wheat flour. Specific bronchial challenge identifies the allergen responsible for asthma, measures the level of sensitization and thus can prevent the occupational exposure. PMID- 9411617 TI - [Giant thymic lipoma: anatomical, radiologic and clinical aspects]. AB - We report the case of a 51-year-old man who presents with worsening dyspnea at exercise caused by a large thymic lipoma (6 kg). We present the clinical, radiological, and spiral CT scan features of this rare and benign tumor and correlate them with the pathologic findings. PMID- 9411618 TI - [Interstitial lung disease linked to fluoxetine]. AB - Fluoxetine (Prozac) is a new anti-depressant introduced on the marked in France since 1989. We report a case of a patient who developed progressive dyspnea without any other symptoms who had been treated with Prozac for nine months. Clinical examination found bilateral basal crepitations. Laboratory assessments showed evidence of inflammation (ESR: 50 mm per hour; CRP 8.9 mg/l), the pulmonary radiograph and thoracic CT scan showed evidence of bilateral diffused interstitial disease. There was also a restrictive ventilatory defect (TLC 59 per cent predicted; FEV1 79 per cent predicted; FVC 70 per cent predicted), effort hypoxaemia and alveolar hyperlymphocytosis (35 per cent). The clinical, radiological and pulmonary function improved and the bronchoalveolar lavage became normal three months after stopping Prozac suggesting a role for this drug in the genesis of this interstitial pneumonia. PMID- 9411619 TI - [Pseudotumorous lung opacities in a patient with AIDS]. PMID- 9411620 TI - [Quality assurance by patient surveys in medical rehabilitation: assessment and evaluation of rehabilitation structures and processes ("satisfaction of rehabilitation patients")]. AB - Since 1994 Germany's statutory pension insurance schemes have been developing a multifaceted programme to assure and optimize the quality of their medical rehabilitation measures. One part aimed at developing a postal questionnaire for patients 8 to 12 weeks after rehab. It comprises two sections: one for the assessment of patients' perceptions and ratings in relation to rehab structures and processes, the other of changes of their health status. The article describes the development of the first part ("rehabilitee satisfaction"). We basically defined relevant sectors and qualities of medical rehab, analysing existing questionnaires, letters of complaint, results of expert interviews and patient focus groups. Mainly based on results from one large study of rehabilitees from two different pension insurance schemes (for blue or white collar workers), we report on psychometric properties of the final instrument, especially its contents, reliability, internal structure, validity and reference values. In February 1997, the instrument was accepted by the association of German pension schemes and may now be used in the routine phase of the quality programme, except for patients with psychosomatic disorders or substance abuse. PMID- 9411621 TI - [Position of the Public Health Working Group of the CDU/CSU legislative faction]. PMID- 9411622 TI - [Position of the SPD legislative faction on health policy (I) and on control in the area of occupational rehabilitation]. PMID- 9411623 TI - [Position of the PDS legislative group on cost saving measures by federal government in the area of rehabilitation]. PMID- 9411624 TI - [Inadequate policy in occupational rehabilitation]. PMID- 9411625 TI - [Reimbursement for medical services in rehabilitation. A contribution to uniform measures]. PMID- 9411626 TI - [Basic considerations for increasing flexibility of services in medical rehabilitation]. AB - Medical rehabilitation measures continue to be delivered nearly exclusively on an inpatient basis. A graded treatment concept comparable to those in place for instance in the acute or long-term care fields, is non-existent, while the medical need for a community-based ambulatory rehabilitation system is uncontested. Not least before a background of ever scacer financial resources among the rehabilitation agencies involved, non-inpatient rehabilitation has been gaining significance as it is generally hoped to achieve greater economy of service delivery. As a logical consequence, the demand for greater flexibility in delivering medical rehabilitation services and benefits is a focus of attention in the current rehabilitation policy debate. Greater flexibility in medical rehabilitation delivery means replacing portions of rehabilitation measures currently provided on an inpatient basis by partial-hospitalization or ambulatory service delivery; or continuing and complementing inpatient rehabilitation measures by non-inpatient ones; or using non-inpatient measures to provide aftercare post-discharge from inpatient rehabilitation. Moreover, ambulatory rehabilitation is intended to reach the population unamenable to attending far away inpatient programmes. It is of utmost significance for the future evolution of ambulatory rehabilitation that the various financially responsible agencies involved in rehabilitation take a joint position on the subject of non-inpatient rehabilitation. Several proposals are submitted in this respect; for indications in the fields of neurology, orthopedics, cardiology, and geriatrics, catalogues of degrees of severity considered suitable for ambulatory rehabilitation are first presented. PMID- 9411627 TI - [Quality assurance in geriatric rehabilitation--approaches and methods]. AB - It did not take the provisions of the 5th Book of the Social Code for quality assurance issues to gain significance in the field of geriatric rehabilitation as well. While in the surgical specialties, experience in particular with external quality assurance have already been gathered over several years now, suitable concepts and methods for the new Geriatric Rehabilitation specialty are still in the initial stages of development. Proven methods from the industrial and service sectors, such as auditing, monitoring and quality circles, can in principle be drawn on for devising geriatric rehabilitation quality assurance schemes; these in particular need to take into account the multiple factors influencing the course and outcome of rehabilitation entailed by multimorbidity and multi-drug use; the eminent role of the social environment; therapeutic interventions by a multidisciplinary team; as well as the multi-dimensional nature of rehabilitation outcomes. Moreover, the specific conditions of geriatric rehabilitation require development not only of quality standards unique to this domain but also of quality assurance procedures specific to geriatrics. Along with a number of other methods, standardized geriatric assessment will play a crucial role in this respect. PMID- 9411628 TI - [Social rehabilitation and earning capacity]. AB - Social rehabilitation, if contrasted with medical and vocational rehabilitation, has hitherto remained a residual category. Vocational resettlement, in conjunction with medical outcome, being the central criterion of successful rehabilitation, it is only on failure in that respect that social rehabilitation aspects come into view. Developments in the society at large, such as changing values, individualization and differentiation of life styles, as well as mass unemployment, working world rationalization, a fragmented labour market and a trend towards shorter working lives, however, warrant renewed attention to the relationship among vocational and social rehabilitation. The problems involved in returning to work after coronary bypass surgery serve to show that occupational resettlement is strongly dependent on the work orientation a patient has developed over his life course as well as on the overall socioeconomic conditions at hand. The ICIDH and its concept of social handicap provides a useful point of departure in seeking to develop a theoretical interpretation of these findings and to expand them towards a systematic concept of social rehabilitation. PMID- 9411629 TI - [Participants of mother-child health resort treatments]. AB - Mother-child health-resort treatments have for many years been a major component of rehabilitative and preventive health care for women. The rising significance of health-promotion measures on the one hand, and increased attention to women's health aspects in rehabilitation and prevention on the other, have made in-depth consideration of the issues involved in target-groups, participation in and conceptual potential of mother-child programmes necessary. In a questionnaire study of n = 103 women participating in a mother-child programme, data were collected concerning their life situation, life satisfaction, psychosocial burdens and psychovegetative complaints, the expectations and needs hoped to be met by the preventive and rehabilitative programme features, as well as the motives for participating. The findings show that the life situation of the women in our study is characterized by a high degree of stress and strain, by physical complaints and dissatisfaction with life. The women clearly state the relationships they see among their current life situation, their subjective emotional state, their health behaviours, and their subjective physical status. Programme participation, on the one hand, has to do with needing a rest and easing of tension; yet on the other hand, the responding women also express a desire to confront current life themes, to find room for reconsidering attitudes, behaviours and the potential for change. Implications for shaping women-specific rehabilitative and preventive programmes are formulated on the basis of the study findings. Future studies in the field of mother-child health resort programmes should be directed at evaluating the treatment programmes available. PMID- 9411630 TI - [Competence in verbal communication of deaf adults in Austria. I: On the intelligibility of spoken language]. AB - Intelligibility of spoken language of deaf adults was assessed by hearing persons. The test procedure was intended to get as close as possible to an authentic communication situation. 5 of 6 deaf speakers are not intelligible to naive listeners. Furthermore, the listeners tend to highly overestimate their understanding of the meaning of the utterances heard. Distorting written reproductions are frequent. There are significant differences between the ability of comprehension of each of the hearing persons. The necessity of visual communication systems and -helps is stressed by the deficits in spoken language found by the study at hand. PMID- 9411631 TI - [Competence in verbal communication of deaf adults in Austria. II: Vocabulary]. AB - The understanding of isolated written words by deaf adults was checked using a basic list of 500 German words developed for this study. Even at this basic level about 7% of the words could not be identified by the deaf subjects. There were particular problems with words from colloquial German, function words, terms without mouthing in sign language and cover terms. Concerning rehabilitation the use of sign language vocabulary already known by the deaf for written language vocabulary training and a stronger orientation towards functional everyday communication is recommended. PMID- 9411632 TI - [Subcutaneous apomorphine in Parkinson disease]. PMID- 9411633 TI - [Nasal obstruction]. AB - When we talk about nasal obstruction, we should define this notion by differentiating between a nasal obstruction, linked to the presence of a physical or inflammatory obstacle on the level of the nasal passages, and the feeling of a nasal obstruction or breathing discomfort described by the patient. The ear, nose and throat explanation will have to take an interest in all the possible causes. The methods of investigation associate anamnesis and clinical examination, helped by endoscopes or fibroscopes, immunological and allergological examinations, computed tomography and magnetic resonance imaging, rhinomanometry and less often use examinations such as muco-ciliary examinations, nasal cellular examinations or the examination of the sense of smell. Nasal obstruction can also be caused by anomalies in the structure of the nasal passages or paranasal cavities, or by anomalies in the mucosa in the nasal passages, or by tumours. Subjective nasal obstruction or nasal discomfort is a pathology at the limit of the conscious and subconscious whose working bases are still not fully explained. Treatment of nasal obstruction can be medical or surgical, but it must absolutely take into account the aetiology and not just the symptoms. PMID- 9411634 TI - [Sudden deafness]. AB - Sudden deafness is a real clinical entity. Very important advances have been achieved these last years in cochlear physiology and afford a better knowledge of the mechanisms of deafness but not the causal agent. Two major pathogenic hypotheses are formulated: viral agent and vascular cause. Two peaks of prevalence in relation with age are described: one between 30 and 40 and the second between 55 and 60. Sudden deafness is a sensorineural emergency. The treatment primarily aims at the restitution of hearing by minimizing the period of cellular ischaemia. PMID- 9411635 TI - [Non-pharmacologic treatment of arrhythmia: what the general practitioner should know]. AB - Important advances have been made in the field of supraventricular and ventricular tachycardias. Catheter ablation procedures are potentially curative in many patients suffering of supraventricular tachycardias. The technique uses radiofrequency current. This form of energy is already very familiar to surgeons. Expected benefits as well as potential complications have to be discussed with the patient prior to the ablation procedure. The automatic internal defibrillator has been commercially available since the end of the eighties. It delivers electrical therapy in case of ventricular malignant arrhythmias, for instance ventricular fibrillation. The sudden death rate is markedly reduced in implanted patients. The long term prognosis however remains critically dependent of the left ventricular function and possible progressive heart failure. Reimbursement depends on pre-implant agreement by the Social Security. At the present time, no reimbursement is provided for prophylactic indications. These two techniques facilitates the treatment of patients previously described as "resistant cases". PMID- 9411636 TI - [Cardiac pacing in general practice]. AB - Cardiac pacemakers are used to maintain a sufficient cardiac rythm. The last generation may stimulate and sense both cardiac chambers, atrium and ventricule (physiologic pacemaker or dualchamber pacemakers). The rythm may also be adapted to the patient's activity (mode rate-adaptative or sensor-driven). All the pacemakers are multiprogrammable and a follow-up program in a pacemaker clinic is mandatory as every programmable parameter and pacing mode is adapted individually to the patient and also because complications are not uncommon. Despite high protection, pacemakers may be susceptible to certain sources of electromagnetic interferences. The maximal risk is present in medical environment and may easily be prevented (magnetic resonance imaging, defibrillation, electrosurgical cautery ...). PMID- 9411637 TI - [Electrocardiography and case reports]. PMID- 9411638 TI - [The general practitioner and abnormal liver function tests]. AB - In case of abnormal liver function tests, it's necessary to distinguish different situations, starting from this first data. We will successively consider: the high and moderate acute increases of aminotransferase, the chronic increases of aminotransferase, the isolated cholestase picture and the isolated increases of gamma GT or of bilirubine. We will finish with a partial survey about drug induced liver diseases. PMID- 9411639 TI - [How to follow treated and untreated chronic hepatitis B and C]. AB - Viral hepatitis B and C are important causes of chronic liver disease, cirrhosis and hepatocellular carcinoma. Indications for the treatment of these two forms of chronic viral hepatitis are outlined as well as the practical modalities for the follow-up. PMID- 9411641 TI - [New viral hepatitis]. AB - Viral hepatitis are essentially caused by 5 types of viruses which differ in way of transmission and their evolution to chronicity or not. Like the virus causing hepatitis A, the E-virus-discovered en 1983-is a virus with oral-fecal transmission, responsible only for acute hepatitis which may be fulminant, notably in pregnant woman. Responsible for epidemics in Asia and Africa, the E virus is nearly non-existent in our regions. Just like the B, C and D viruses, the G-virus is a RNA-virus with intravenous transmission. Notwithstanding a high prevalence, its pathogenic role remains hypothetical so that some hesitate to consider it as a virus causing clinical hepatitis. Etiological viral or non-viral agents for the cryptogenic hepatitis which can appear as acute, fulminant, post transfusional or chronic illness, remain to be discovered. PMID- 9411640 TI - [Hepatitis A and B vaccines]. AB - Hepatitis A is a beginning disease in childhood but sometimes a very dangerous disease in adults. It is necessary to vaccine the non immune "risk groups". Hepatitis B is responsible for two millions deafs every year. Hepatitis B is a sexually transmissible disease. Everybody would be vaccinated. PMID- 9411642 TI - [Optimal therapeutic strategies in ovarian epithelial cancer in 1997]. AB - "Optimal" chemotherapy for advanced ovarian cancer has constantly evolved over the last 2 decades through the conduct of prospective randomized clinical trials. Because 3 such important trials have recently disclosed provocative results there are reasons to believe in the emergence of new "standard" treatment approaches for this disease towards the end of this century. 1) The Intergroup trial found a survival advantage for intraperitoneal cisplatin as compared to intravenous cisplatin following optimal debulking surgery. 2) In the EORTC-GCCG trial, which recruited patients with bulky disease at completion of primary surgery, survival was prolonged when interval debulking surgery was performed after 3 cycles of chemotherapy. 3) Paclitaxel-cisplatin was associated with a marked survival advantage in comparison with cisplatin-cyclophosphamide in the GOG trial, which enrolled suboptimally debulked patients. These trials clearly have important implications for the future management of ovarian cancer patients and from a health economy point of view: for these reasons, two of them (2 + 3) have been repeated by other groups, and results of these confirmatory trials should be available soon. There are a number of new treatment options for "Platinum resistant" patients including docetaxel, topotecan, gemcitabine, oxaliplatin: but none of them is "optimal". An active search for new drugs in this setting remains a high priority. Finally, with the expanding knowledge of the molecular biology of cancer in general and ovarian cancer in particular, one can now start thinking of new molecular targets for treatment intervention including transmembrane tyrosine kinase growth factor receptors, matrix metalloproteinases, the vascular endothelial growth factor and so on.... PMID- 9411643 TI - [Current topics in pulmonary oncology]. AB - Thoracic oncology practice is changing with the end of the century. New diagnostic tools like photodetection allow to diagnose at a white light undetectable level. Preneoplastic lesions or very early cancers that can be locally treated by photochemotherapy, cryotherapy or brachytherapy. The natural history of lung cancer will also be better known. Concerning advanced disease, cisplatin chemotherapy improves survival of patients with stage III non-small cell lung cancer in combination with surgery or chest irradiation and of those with stage IV in comparison with best supportive care alone. New approaches in small cell lung cancer seem promising like accelerated chemotherapy, early chest radiotherapy and maintenance treatment. Moreover, a series of new active cytotoxic agents has been recently identified. The complexity of these modalities makes more and more necessary a integrated pluridisciplinary approach of the lung cancer patient. PMID- 9411644 TI - [Colorectal cancer: a controllable disease]. AB - Colorectal cancer is one of the most frequent cancers in western countries. Five years after curative surgery and adjuvant chemotherapy, about 10% more patients with Dukes C colon cancer are free of disease compared to the control group. Several regimens using 5-fluorouracil with folinic acid (5FU/FA) or levamisole are available. For patients with disseminated disease. 5FU/FA based treatments allows a doubling in survival as compared to best supportive care. Moreover, quality of life is significantly improved. New agents are upcoming. Tomudex seems to be equivalent to 5FU/FA but easier to administer. CPT-11 or oxaliplatin have been investigated in second line after failure of 5FU/FA. The available data suggest that median survival is prolonged by ten months, in addition to what was already obtained in first line with 5FU/FA. Colorectal cancer must be revisited. An active approach of our patients should allow to reduce the incidence of the disease, to increase the number of disease free patients 5 years after surgery, to prolong survival and improve the quality of life of those who will develop metastatic disease. PMID- 9411645 TI - [Progress in radiotherapeutic approaches to anal, rectal, bronchial and prostatic cancers]. AB - During the last years, various progress has been made in the irradiation of cancers: combined treatment modalities with surgery and chemotherapy, the radiation technique itself (conformal radiotherapy, brachytherapy ...), alterations of the radiation fractionation, the use of radiosensitizers. These modifications will be illustrated through the treatment of anorectal, lung and prostatic cancers. PMID- 9411646 TI - [The ambulatory cancer patient treated by chemotherapy]. AB - The success of cancer chemotherapy in ambulatory patients depends on the perfect knowledge of the various side effects of the drugs and moreover on a close cooperation and comprehension between the medical staff, the family practitioner and the patient. Quality of life is one of the most important aims of the treatment in oncology, especially in an ambulatory setting. PMID- 9411647 TI - [The ambulatory cancer patient treated by radiotherapy]. AB - Without a sufficient knowledge of the acute and late normal tissue reactions due to radiotherapy, a general practitioner is sometimes helpless and hopeless, facing a patient consulting with questions, or side effects. How to manage a skin reaction, a mucositis, a leucopenia, a hair loss, ...? The aim of this paper is to describe the pathogeny of frequent acute and/or late reactions related to ambulatory radiotherapy, allow an early diagnosis, facilitate a follow-up in common and provide guidelines for the symptomatic medical treatment. PMID- 9411648 TI - [Falls in elderly persons: evaluation of risk and prevention]. AB - Falling represents a common and dangerous problem for the elderly. Approximately 30% of persons over age 65 who are independent and living on their own will fall each year. Falls in this age group result in a bone fracture 5% of the time. Risk factors for falls involve both environmental hazards and host issues. Evaluation of the patient, his or her living conditions, and any personal unsafe behaviors can identify those at risk of falling. Efforts to prevent falls in the elderly have involved education, strengthening exercises, medication evaluation, and environmental improvements. PMID- 9411649 TI - [Evaluation of dependence in the aged]. AB - On account of the ageing of the population, the general practitioner will be more and more solicited to care for old people. The determinations of the degree of dependence of the elderly person is more a global bio-psycho-social concept than the classic view of the curative medicine. Two scales of evaluation of the state of dependence--the Katz scale and the Old People Aid scale--are commonly used in Belgium. Are they valid and do they reflect the actual recourse for aid and care? To answer the questions, this article refers to a study realized in the french community of Belgium about people older than 65 years. The analysis of the dependence degree will allow a better definition--in respect of the patient's will and choice--of the objectives and needed resources. PMID- 9411650 TI - [Leg ulcers and bedsores--introduction]. PMID- 9411651 TI - [Clinical aspects of leg ulcers]. AB - Clinical aspects of leg ulcers are reviewed. Among the most important factors to be kept in mind are: localisation, features of the ulcers and surrounding skin, clinical history and general physical examination. Leg ulcers are most frequently caused by vascular diseases. PMID- 9411652 TI - [Particular aspects of ulcers in children]. AB - Leg ulcers of juvenile onset are uncommon. The infectious origin is fairly frequent but the presence of leg ulcers in children should prompt an investigation into possible underlying causes especially hemangioma, vasculitis, inborn errors of metabolism (i.e. prolidase deficiency), hemoglobinopathies, occult spinal dysraphism and immunodeficiencies. Bacteriological investigations are essential and a skin biopsy specimen may be able to differentiate some of these disorders. PMID- 9411653 TI - [Physiopathology of leg ulcers]. AB - Leg ulcers occur more commonly in the elderly and represent in our ageing western countries a real public health problem. Different mechanisms can induce these cutaneous ulcerations. An underlying vascular insufficiency, venous, arterial or mixed can be demonstrated in most cases. The management of leg ulcers needs an identification of their cause(s) and a good knowledge of their physiopathology, in order to ensure adequate treatment. The purpose of this presentation is to remind of the diversity of physiopathological mechanisms involved in the onset of these ulcers. PMID- 9411654 TI - [Diagnosis and general treatment of leg ulcers]. AB - A comprehensive history and the physical examination are the mainstays of the diagnosis of leg ulcers. Some diagnostic procedures will allow to establish a proper diagnosis in most cases. The precipitating cause of ulcer has to be taken in charge if possible. Most leg ulcers are secondary to chronic venous insufficiency: the principal treatment modalities are compression and increasing venous return by adapted exercises. The evidence for the efficacy of medications is still limited. PMID- 9411655 TI - [Local treatment of leg ulcers]. AB - Local treatments of leg ulcers require necrotic tissues destruction and preventing and reducing surinfection with local antiseptics. Local and general antibiotherapy are proposed in defined circumstances. Choice of local dressings are dependent on necrosis, degree of exustion, global cost and compliance of the patient. PMID- 9411656 TI - [Surgical closure of leg ulcers]. AB - Surgical closure of leg ulcers has to be preceded by treatment of their etiologies in order to avoid recurrences. Best coverage technique is achieved with the use of a meshed split thickness skin graft, harvested with a dermatoma. Skin graft take depends on the vascular quality of the recipient bed, on the technique used and also on the post-operative care. PMID- 9411657 TI - [Physiopathology of bedsores]. AB - Pressure is the primary pathogenic factor in the development of decubitus ulcers. Other major factors are shearing forces, friction and moisture. Significant intrinsic risk factors are immobility, age-related diseases, nutritional status, medications and smoking. The morbidity and mortality related to the complications of pressure sores are quite significant. Prevention is essential and is best achieved by identification of high risk patients. The therapeutic approach is based on the grade of pressure ulcer. PMID- 9411658 TI - [Pressure sores: management and treatment]. AB - The global management of pressure sores is best ensured with a multidisciplinary approach. We present the experience of the "Groupe de Travail Escarres" (Pressure Sore Workgroup) which gathers physicians and nurses interested with this pathology. The majority of the decubitus ulcers will heal spontaneously with a conservative treatment only. This treatment typically aims at relieving the causes that lead to pressure sores, at eliminating the necrotic tissues, at obtaining favourable local conditions to allow wound healing and at controlling the health status of the patient. Surgical treatment of pressure sores is indicated when wound healing does not occur and when the health status of the patient is sufficiently good. Defect coverage is best carried out using myocutaneous flaps since their excellent blood supply allows a good cleansing of the wound. PMID- 9411659 TI - [Growth hormone treatment in adults: legitimate and illegitimate indications]. AB - Growth hormone replacement therapy of hypopituitary adult patients has demonstrated its usefulness during the last 12 years: it decreases excess body fat while raising lean mass and basal metabolic rate, it ameliorates the lipid profile of the patients and decreases their peripheral vascular resistance, it increases bone mineralization and ameliorates the quality of life of treated individuals. Based on these results, many other applications of growth hormone have been developed: treatment of old age, of simple and android obesities, of osteoporosis and extreme catabolic situations. The data collected have been of great pathophysiological interest, but do not lead for the present towards therapeutic indications. PMID- 9411660 TI - [Alcoholism: relapse prevention]. AB - Prevention of alcoholism relapse implies an early detection of the problem (abuse or dependence). This can be achieved through clinical interview, biological evaluation, psychological assessment (i.e. "The Drinking Habits"--I. Pelc) or according to the "Alcoholism Decision Tree" (The Plinius Maior Society). In order to prevent alcoholism relapse, strategies to control the clinical symptoms related to the "Post-Detoxification Syndrome" have recently been highlighted. Further are discussed the benefit of various psychotherapeutic modalities as well as psychopharmacological interventions. According to a better knowledge of the neurochemical and neurobiological basis of alcoholism and craving, the contributions of specific psychopharmacological agents active on the dopaminergic, serotoninergic, glutamatergic systems as well as blocking of opiate receptors are discussed. Acamprosate, as a specific modulator of the glutamatergic system, recently marketed in Belgium, appears one of the most promising new pharmacological agent in relapse prevention when the patient benefits of psychosocial support. PMID- 9411661 TI - [The anti-leukotrienes: their use in asthma]. AB - Inflammation in asthma plays a predominant role in the genesis of bronchial obstruction and hyperreactivity. Treatment of bronchial inflammation since the early stages of asthma has become an essential element of therapeutic strategy and rests chiefly on inhaled glucocorticoids. However, inhaled glucocorticoids pose some problems in terms of inhalation technique, compliance, local tolerance and, to a lesser extent and at high doses, of systemic tolerance. Antileukotrienes represent a new class of potential anti-asthma drugs. They act either by inhibition of 5-lipoxygenase, a key enzyme in the leukotriene (LT) synthesis pathway, or by blocking LT receptors. The LT antagonists developed in asthma are mainly specific for a receptor subtype, the CysLT1 receptor, involved in the bronchoconstrictor and inflammatory effects of sulfidopeptide leukotrienes (LTC4, D4 and E4). Two antagonists of CysLT1 receptors have been recently launched on the market: ONO-1078, pranlukast, Onon and ICI-204,219, zafirlukast, Accolate. The CysLT1 antagonists cause bronchodilatation with a beta 2-agonist additive effect, confer effective protection in bronchial provocation studies and particularly, they improve clinical scores, pulmonary function and beta 2-agonist rescue inhaler use. Additional studies, especially comparative with respect to reference treatments, mainly inhaled glucocorticoids, will be required to define their place in the global strategy for asthma management. PMID- 9411663 TI - ["Doctor, my knee hurts...", the generalist's point of view]. AB - Knee pain is a relatively common symptom in family practice. It is occurring at the rate of 48 fo 1000 patients/year in the Dutch general practitioner's non selected population. The different investigation techniques are listed and studied, leading to a global classification (non inflammatory (degenerative, mechanical or traumatic), inflammatory or septic knee), then to a more specific and more operational one for an optimal therapeutic way. PMID- 9411662 TI - [Post myocardial infarction management]. AB - Post myocardial infarction preventive strategies are effective in reducing morbidity and mortality. Life threatening risk factors can be controlled by medication in addition to dietary measures. The following update will be proposed on: new lipidic and non lipidic risk factors; necessity to control risk factors, nicotine dependence and lipid metabolism disorders in particular; patient follow up; drug treatment efficacy (beta-blockers, aspirin, ACEI, statins); compliance to a varied drug treatment regimen. Three newly developed treatments will be presented: the first sartan, clopidogrel and troglitazone. PMID- 9411664 TI - [Clinical examination of the knee]. AB - The clinical examination of the knee is very important. After listening to the patient, one asks him various questions and one performs the clinical examination. After that only it is possible to choose the best examinations to be done, if necessary. The various aspects of the interrogation and the clinical examination are described according to the different structures of the knee which may be injured. PMID- 9411665 TI - [A "good choice" of diagnostic imaging examinations: the knee]. AB - A medically sound and cost-effective choice of diagnostic imaging investigations in knee pathology must be supported by a good evaluation of the respective strong and weak points of the available methods. These are: conventional radiology, C.T., arthrography, arthro C.T., ultrasound, scintigraphy and magnetic resonance imaging. Based upon that analysis, decisional algorithms are proposed, in function of the clinically suspected pathologies. PMID- 9411666 TI - [The outcome of the patient treated with methadone]. AB - The treatment of drug addiction by methadone is questioned: there is a shift from "all of abstinence" to "all for risk limitation". There is a modification of the clinical relationship, due to specific resources allocated for methadone treatment, modifying also the connections between medicine, Public Health and Public Security. PMID- 9411667 TI - [Child abuse and medical confidentiality]. PMID- 9411668 TI - [What is your diagnosis? Necrotizing fasciitis of the left neck area]. PMID- 9411669 TI - [Molecular tumor genesis of colorectal carcinoma]. PMID- 9411670 TI - [Colon carcinoma: molecular diagnosis and therapy]. AB - Colorectal cancer (CRC) is one of the most frequent cancers in Western countries. The identification of individuals at risk and the early diagnosis of CRC are of critical importance since a large proportion can be prevented or cured by surgical removal before metastasis has occurred. With increasing understanding of the genetic basis of hereditary and sporadic (non-hereditary) CRC, it becomes feasible to detect genetic alterations by molecular techniques. Familial adenomatous polyposis (FAP), hereditary nonpolyposis colorectal cancer (HNPCC) as well as early stages of spontaneous CRC can be diagnosed by molecular characterization of the adenomatous polyposis coli (APC) gene, the ras oncogene and other genes, respectively, in DNA from peripheral blood, stool or intestinal biopsies. At present, careful patient and family history, physical examination, testing for occult blood as well as colonoscopy are still the key elements, however, for clinical patient management. Molecular diagnosis will hopefully soon complement these analyses and should result in a reduction of morbidity and mortality from CRC. Further, gene therapy offers some potential to prevent or treat CRC. PMID- 9411672 TI - [Adjuvant and palliative immunotherapy of colorectal carcinoma]. AB - A review of the clinical trials on immune therapies in cancer reveals that this approach can, in rare cases, induce complete remissions in individual cancers. Since these trials have usually involved patients with large tumor masses, tumor cell inaccessibility is probably a major reason for the prevailing failures. Minimal residual disease, the stage when tumor cells are few and dispersed, should therefore be a more promising target for immunological approaches. This hypothesis is supported by a prospective randomized trial on patients with resected Dukes C colorectal carcinoma. 189 patients were assigned to an observation regimen or to postoperative treatment with 500 mg of 17-1A antibody, followed by four 100 mg infusions each month. After a median follow-up of five years, antibody treatment reduced the overall death rate by 30% and decreased the recurrence rate by 27%. The effect of antibody was most pronounced in patients who had distant metastasis as first sign of a relapse, an effect that was not seen for local relapses. Thus in addition to strategies designed to produce more effective reagents, efforts need to be concentrated on directing immunological effectors towards the appropriate target. PMID- 9411671 TI - [Current aspects of adjuvant and palliative chemotherapy in colorectal carcinoma]. AB - With an annual incidence rate of 30 to 40 per 100,000 colorectal carcinoma is the second most frequent malignancy in Germany. Despite the poor outcome of patients suffering from this disease important advances have been made in the standardisation and improvement of palliative and adjuvant treatment in patients with colorectal cancer. For the systemic chemotherapy 5-fluorouracil (5-FU) remains the most important cytotoxic agent and biomodulation of the therapeutic activity of 5-FU with methotrexate or particularly folinic acid has been clinically established, yielding response rates in 20 to 35% of patients. Current investigations of systemic treatment are aiming into three directions: 1. investigation of high-dose continuous (24-hours) 5-FU application (with or without modulation by folinic acid); 2. evaluation of new, effective cytotoxic agents, among which the camphotecin derivative CPT-11 (irenotecan) and the specific thymidilate synthase inhibitor Tomudex appear to be the most promising drugs as single agents and/or in combination with 5-FU; 3. use of orally available fluoropyrimidine derivatives with high bioavailability which may substantially improve the quality of life in palliative therapy. The postoperative adjuvant treatment of patients with Dukes C colorectal cancer is established clinical practice and the combination of 5-FU and levamisol given for one year will result in an improved overall survival of about 15% at five years compared to surgery alone. Although this regimen remains the current standard treatment, alternatives for the adjuvant treatment may be the use of 5-FU and folinic acid given for only half a year post surgery, locoregional perfusion of the liver with 5-FU alone via the portal vene by 7-day continuous application or the use of 17-1A monoclonal antibody immunotherapy after curative resection. Further improvement may be achieved by the combination of immunotherapy and chemotherapy which is currently tested in clinical studies. Future recommendations for the adjuvant treatment of colorectal cancer will not only be based on therapeutic efficacy but will also have to take costs of treatment into account. Better definition of high-risk patient groups for adjuvant treatment is needed. PMID- 9411673 TI - [After-care of colorectal carcinoma: what has efficacy?]. AB - The efficacy of standardized follow-up examinations after surgery for colorectal carcinoma has been repeatedly questioned. Although many studies have assessed the value of different diagnostic tools none of these procedures proved to have a predictive value high enough to accurately predict the recurrence of disease and to justify its regular use in these patients. Even a combined diagnostic approach provided a benefit only for a minority of these patients (3.5 to 4.5%). Considering the physical and psychological strain imposed by this approach risk adapted follow-up schemes are urgently needed. Prognostic parameters indicating the individual risk for disease recurrence can be deduced from increasing CEA values as well as from classification and grading of the primary tumor. Colonoscopy is an effective procedure for early detection of intraluminal relapse or metachronous tumors with potential impact on survival of the patient. An abdominal ultrasound study appears to be the method of choice for detection of metastasis of the liver due to its high sensitivity and low invasiveness. Other imaging procedures are not indicated in routine follow-up for colorectal carcinoma. It remains to be demonstrated whether molecular biology or new scintigraphic techniques will be helpful in follow-up examinations of patients with colorectal carcinomas. PMID- 9411674 TI - [HIV replication in labile equilibrium]. PMID- 9411675 TI - [The outcome of Alzheimer's disease]. PMID- 9411676 TI - [Psychiatric disorders in Alzheimer's disease and organization of care]. AB - The organization of care for a patient suffering from Alzheimer's Disease (AD) must consider the intricate psychiatric and cognitive problems associated with this disorder. It will be complementary to the primary care given by the patient's relatives and offer a large choice of care structures including different possibilities between the patient's home and long term psychogeriatric facilities. It should be flexible and favour transitory solutions. It has to adhere to a basic philosophy which takes into account the patient's personal history, focusing on his behavior seen as the result of his efforts to adapt. Finally, the organization needs to assume, at all care levels, an approach of psychological and psychotherapeutic support to the patient and his family. PMID- 9411677 TI - [Circadian rhythm disorders, motivation and Alzheimer's disease]. AB - In Alzheimer's disease, when the breakdown of the sleep's circadian rythm is sudden, it reflects frequently a concomitant somatic or psychiatric disorder. If the trouble appears progressively, there is less pathological or disturbing behaviors in the beginning. Later, it is often associated with a loss of interest for the daily living activities (perte de motivation). PMID- 9411678 TI - [Motor performance in Alzheimer's disease]. AB - Movement and motor expression are indirect means of approaching the affectivity of patients suffering from Alzheimer's disease. This is why it is very important to describe here two therapeutical approaches: a) on the one hand, an approach which focuses on the physical and psychical functioning of the patient: b) on the other hand, an approach which validates a new type of therapy, psychomotor therapy. PMID- 9411679 TI - [Affective disorders in Alzheimer's disease]. AB - The links between depressive syndrome and Alzheimer's disease are problematic concerning the diagnosis and the therapeutic choice. The concepts of pseudo dementia and late onset depression are shortly discussed. The hypothesis concerning the relationship between depression and Alzheimer's disease are summarized. The clinical data relevant for the diagnosis and the therapeutic guidelines are discussed. PMID- 9411680 TI - [Phenomenology and treatment of delirium in Alzheimer's disease]. AB - Meaning of psychiatric disorders in Alzheimer' disease has been underestimated for a long time. Among these disorders, delusional states represent the most frequent causes of such phenomenon after depressive disorders. It's probably of importance to distinguish delusional and paranoid ideation from the missidentidentification phenomenon. Neuroleptic treatment contribute to improve the quality of life of these delusional patients. PMID- 9411681 TI - [Psychodynamic treatment in Alzheimer's disease]. AB - The psychodynamic approach to a patient's experience of having Alzheimer's disease requires more research. It is a therapeutic objective to allow a patient with Alzheimer disease to obtain a realistic perception of his own condition, particularly since this is frequently unusual for this group of patients. PMID- 9411682 TI - [Alzheimer's disease and milieu therapy]. AB - Frequency of dementia increases quickly with age. Such syndromes touch 2-3% of people between 65 to 69 of age, but 20-30% of the people after 85 of age. To treat the patients, early diagnosis is very important. Therapeutic measures taking in account the environment, the pharmacotherapy and prevention of malnutrition is of growing importance. PMID- 9411683 TI - [Body-mediated therapies in geriatric psychiatry]. AB - The use of body mediated therapies at the Clinical geriatric psychiatry became possible due to the convergence between multi, inter and transdisciplinary approaches and the clinical experience in this application of various therapeutic technics. This approach at the aged clinic must take into consideration the specificity of the correlation to the body of the aged person beside the advantages and the needs that resent this approaches to both the patient and his therapist. PMID- 9411684 TI - [Behavior disorders of neurological origin]. AB - The modifications of behavior related to neurological diseases are various and important to be correctly diagnosed. The purpose of this article is to present the clinical features of main neuropsychiatric syndromes: depression, delusions, anxiety, obsessive-compulsive syndrome. The differential diagnosis is also developed. The appropriate treatment is discussed. PMID- 9411685 TI - [Neurodegenerative disorders and functional imaging: role of astrocytes]. PMID- 9411686 TI - [Neurodegeneration and epileptic crises in Alzheimer's disease mesio-temporal sclerosis]. PMID- 9411687 TI - [Atypical cases of acute headaches of cerebrovascular origin]. AB - Acute headaches can be an important signal of a cerebrovascular event. In some cases, as illustrated in this article, such headaches may have the same characteristics as migraine or be the main feature of unusual types of cerebrovascular disease. Headache should not be underestimated since misdiagnosis of cerebrovascular disease can lead to serious consequences. PMID- 9411688 TI - [Comparison of elderly persons nutritional status living at home, in an institution, or in a semi-rural hospital]. PMID- 9411689 TI - [New treatments, patient autonomy, (always) limited resources. Ethical aspects in the management of patients with HIV/AIDS]. PMID- 9411690 TI - [Paternity testing expertise]. PMID- 9411691 TI - [Legal medicine is a way of thinking]. PMID- 9411692 TI - [Sexuality experience of patients living with HIV: literature review]. PMID- 9411693 TI - [Psychological and psychodynamic education of physicians]. AB - Not every Doctor needs a training in the doctor-patient relationship and in the knowledge of the psychodynamics issues of his patients. But it is a must for the doctors who treat and look after their patients. The people in charge of this training should consider the importance of the unconscious and consequently of the psychoanalytical dimension. The resistances against the psychodynamic approach by the reduction to the common sens, the use of the parallel medicines or the systemic and cognitive approaches. The need and desire of this type of formation in relation to the professional personality of the Doctor. PMID- 9411694 TI - [Passport to the operating room]. PMID- 9411696 TI - [Reconstruction of orbital floor fractures using autologous materials]. AB - Bone and Cartilage Autograft gather all the necessary qualities for an interpositional material to be used in the fractured orbital floor reconstruction, leading to the binocular vision recovery and in term of tolerance. The initial material choice depends on the clinic and orbital tomodensitometry datas, but the final decision is made on the operating findings. However, schematic indications can be drawn up, depending on material characteristics, curve, rigidity and resorption degree, and fracture particularities. PMID- 9411695 TI - [Use of iterative expansion in the early surgical treatment of complex nevi of the head in children]. AB - Extensive of congenital pigmented nevi to the face in an infant is an indication for early exeresis to prevent the risk of degeneration. Search for the best esthetic result has led many authors to healthy skin to a maximum, often relying on tissue expansion. The aim of this study was to present the combination of two expansion techniques, prosthetic expansion and differed natural expansion. Five infants with congenital pigmentary nevi extending to more than 50% of a facial anatomic unit were treated. Total treatment was achieved in all patients with three or four procedures. By combining different expansion techniques early treatment can be proposed with good esthetic results and moderate cost. PMID- 9411697 TI - [Lingual osteoma. Apropos of a case]. AB - The authors present a case of osteoma of the tongue. This tumor was discovered without any functional symptom. Surgical resection with histological examination gives the diagnostic. This choristoma is rare, benign and clinically poor. Osteoma of the tongue is a heterotopic bone tissue and his main differential diagnosis are lingual thyroid and thyroglossal cyst. There are several opinions concerning the histogenesis of this choristoma, but an embryological anomaly is probable. Surgical removal of the tumor is curative, without any reported local recurrence. PMID- 9411699 TI - [Implant-supported prosthesis. The significance of endosseous and peri-osseous implants]. AB - The new extra-oral of Farmand (EPITEC system) is a plate-implant. This concept is really different from the Branemark's implant and the use of EPITEC system give the authors opportunity of a first assessment. It seems that if, of course, Branemark system is still the reference, the EPITEC system certainly has a real interest in thin osseous implantation areas. PMID- 9411698 TI - [Infra-orbital nerve block in early primary cheiloplasty]. AB - A prospective study have studied the effect of infraorbital block during anesthesia in infants with a cleft lip. The study was conducted during the year 1994, in the hospital Necker Enfants-malades. During this period 51 cleft lip surgery were performed. Anesthesia for infants with cleft lip using bilateral infraorbital block is a safe, simple and quick technique, and result in a good longlasting analgesia, seems to decrease the risk of respiratory depression, and allows an immediate and comfortable awakening. PMID- 9411700 TI - [The management of extra-oral implants. Anachronisms and paradoxes]. AB - Used in France for ten years and in Sweden since 1977. Extra-Oral Implants (EOI) represent one of the new therapeutic approach of facial defects treatment by maxillo-facial prostheses fixed on implants and of some transmission deafnesses. Audiologic applications are free of charges for the patient but not the prosthetic applications; the authors note this paradox and some other anachronisms. Finally the authors wonder why Extra-Oral implantology is a "correct" science and why Intra-Oral Implantology remains absent from the Administrative texts as if it was a sort of doubtful therapy without scientific principles. PMID- 9411701 TI - [Intramuscular capillary-venous angioma extending into the infratemporal fossa. Apropos of a case]. AB - A rapidly growing tumor in the temporal area extended into the infratemporal fossa. Imaging findings favored a hypervascularized meningo-sarcoma type extracranial tumor, but the pathology report on the surgical specimen confirmed the diagnosis of capillaro-venous angioma. The therapeutic approach was preoperative hyperselective embolization then access via the temporal and infra temporal fossa for simple tumor exeresis. Mature angiomas and access via the infra-temporal fossa are reviewed. PMID- 9411702 TI - [Late results of the surgical treatment of temporomandibular joint ankylosis in children]. AB - Long-term outcome after surgical treatment of temporomandibular joint ankylosis is not always satisfactory in children. Interposition of a temporalis muscle flap is insufficient to ensure good functional results if the surgical procedure is not followed by careful active and passive physiotherapy. The results can be evaluated only after the child has grown. PMID- 9411703 TI - [Diffuse cervical cellulitis apropos of 15 cases]. AB - A homogeneous series of 15 cases of diffuse cervical cellulitis originating in the oropharyngeal area were collected from December 1990 to April 1995. Despite early management, diffusion of the infectious processus was rapid with extension and development of severe mediastinal or pulmonary complications. When diffuse cervical cellulitis is suspected, early and adapted aggressive treatment must be initiated immediately: suppression of the causal factor, surgical exploration with repeated drainage depending on the clinical course assessed on CT scans, antibiotics effective against the aero-anaerobic flora. This aggressive attitude is required to avoid potentially severe morbidity. Anti-inflammatory drugs, taken alone or with unadapted antibiotics was implicated in 11 cases as a favoring factor. PMID- 9411705 TI - [A case of aggressive juvenile fibromatosis of the mandible]. AB - We report a new case of aggressive juvenile fibromatosis (FJA) in a 20 months old girl. It was initially treated by hemimandibulectomy. A secondary reconstruction was performed with a fibular flap at the age of 9 years. FJA is a locally aggressive lesion which does not metastasize. It occurs chiefly in girls during childhood. Radiographs are non-specific (extensive osteolytic lesion). The clinicopathologic diagnosis is discussed with the other mandibular fibrous tumors in children. PMID- 9411704 TI - [Severe diffuse facial cellulitis]. AB - Twenty cases of severe cervicofacial infections among 184 cases of cellulites treated in 1991 and 1992 at the Lille University Hospital were investigated. Outcome was compared with results reported in the literature. Outcomes were similar and emphasized the critical nature of this type of infection which caused 7 deaths in our series and severe anatomic and functional sequelae. All patients (n = 20) were given anti-inflammatory drugs prior to surgery. The correlation between disease severity and use of anti-inflammatory drugs would appear to be an established fact. Criteria predicting severity were identified in order to obtain a score for high or low risk and to better adapt the medical and surgical management strategy. PMID- 9411707 TI - Proceedings of the IV Seminar on Audiology and Phoniatrics. New strategies in the rehabilitation of profoundly deaf children. Bolzano, Italy, December 1995. PMID- 9411706 TI - [Congenital giant blue nevus. Apropos of a case]. AB - A case of a three years old girl showing a congenital giant blue nevus, in the neck and the occipital region, is presented. The giant blue nevus was associated to an important cutaneous nevomatosis. The treatment has consisted of an surgical excision of an surgical excision of the main lesion, and operative continuations have been simple. The literature study showed 11 cases of congenital giant blue nevus and demonstrate the high risk of malignancy, and the importance of an early surgical treatment. PMID- 9411708 TI - [Perioperative gastrointestinal ischemia in abdominal aortic aneurysm operations]. AB - QUESTIONS UNDER STUDY: Surgery of abdominal aortic aneurysms involves a high risk of postoperative complications. It has been suggested that the incidence of postoperative complications is related to the development of gastrointestinal acidotic episodes of the mucosa. The goal of this study was, therefore, to determine the incidence of gastrointestinal acidotic episodes during repair of abdominal aortic aneurysms, and to test the hypothesis that these episodes predict an adverse postoperative course. METHODS: In 49 consecutive patients undergoing elective surgery for repair of an abdominal aneurysm, intramucosal gastric pH (pHi) was measured perioperatively. The length of the surgical procedure, perioperative intravenous fluid intake, use of vasoactive drugs, APACHE-II scores, days with an endotracheal tube in place, days of intensive care and major postoperative complications were prospectively assessed. The patients were assigned to either a group with a pHi > or = 7.35 or a group with pHi < 7.35 measured towards the end of the surgical procedure, and then compared. RESULTS: The pHi decreased from 7.42 +/- 0.09 after induction of anesthesia to 7.37 +/- 0.07 (p < 0.05) during clamping of the aorta, and continued to decrease to 7.34 +/- 0.08 (p < 0.001) towards the end of surgery and on admission to the surgical intensive care unit (mean +/- standard deviation). The percentage of patients with pHi < 7.35 increased from 10% at the beginning of the operation to 55% on admission to the intensive care unit (p < 0.0001). There was no difference in the postoperative course between patients with pHi > or = 7.35 and those with pHi < 7.35 measured after declamping of the aorta. Patients who had a major complication during their stay in the intensive care unit had lower perioperative pHi values than patients without complications (p < 0.001). CONCLUSIONS: Perioperative gastrointestinal acidotic episodes of the mucosa are common during repair of abdominal aortic aneurysms. The perioperative course, however, is not influenced by these acidotic episodes, despite the fact that patients with complications during their stay in the intensive care unit had lower perioperative pHi values. The routine use of pHi measurements during elective surgery of abdominal aortic aneurysms, therefore, is not justified. PMID- 9411709 TI - [External quality control in the medical laboratory: evaluation of 12 ring trials 1994-1996]. AB - The regulations for reimbursement of laboratory tests provide that such tests will be paid for by social insurance institutions only if the laboratories participate in internal and external quality control schemes. Twelve surveys of the external quality assessment scheme of MQ Zurich (Association for Medical Quality Control) were evaluated. We analyzed the imprecision and inaccuracy of the participant results depending on the analytical system or methods used. Furthermore, the number of participants who met the quality criteria published by FKGRAL (Expert Committee for Overall Revision of Analysis List) was determined. The deviations from the internationally recommended or reference methods respectively were within +/-33% for the metabolites and enzymes if native plasma was used as control sample. If lyophilized samples were used, 5 deviations observed were > +/-33% (maximum +99%). For hematologic parameters the deviations were in the range of +/-10%. The CV's were 5.7-17.6% for wet chemistry methods used by the participants and 4.5-15.1% for the other methods. (Cobas Ready, Ektachem, Reflotron, Vision). For hematologic parameters we found CV's between 4.5 and 14.0%. 69-83% of the participants using wet chemistry methods met the FKGRAL criteria, while 86-98% of participants using one of the other system obtained adequate results. The corresponding figure for the hematologic parameters was 83-93%. The nature of the control samples (native samples or lyophilizate) did not influence the number of participants who successfully passed the survey. The study showed that the surveys are an adequate tool for determining participants with inadequate analytical performance, and in many cases the survey results make it possible to propose the necessary educative measures. PMID- 9411710 TI - [3 siblings with identical, rare pneumopathy]. AB - Pulmonary alveolar microlithiasis was found in three siblings. Only the youngest of them, a former smoker, developed endstage lung disease. The other two are asymptomatic with normal lung function despite impressive changes on all chest radiographs. The role of smoking in perpetuating microlithiasis and furthering the progression of this disease is discussed. PMID- 9411711 TI - [Acute life threatening catatonia--clinical significance and therapeutic possibilities]. AB - Malignant catatonia, associated with different somatic and psychiatric disorders, is a rare, life-threatening syndrome. Immediate recognition and adequate treatment are essential and may be life-saving. We describe a case of malignant catatonia and discuss the clinical implications. Additionally, we review the recent literature regarding epidemiology, nosology, current pathophysiological concepts, differential diagnosis, and treatment recommendations. PMID- 9411712 TI - [Differential diagnosis of the air-fluid level in thoracic roentgen image]. PMID- 9411713 TI - [Why does even high quality psychosomatic research produce so little?]. PMID- 9411715 TI - [The language of medicine in Switzerland 1920 to 1995]. AB - AIM OF STUDY: It is generally accepted that since the end of the Second World War, English has become the main language in the medical field in Switzerland, but scarcely any objective data are available on the development of this process in this country. The aim of the present study was to analyze the frequency of the different languages in the literature references in articles published in the Swiss Medical Weekly over the past 75 years, with special attention to the possible differences existing between articles originating in German-speaking Switzerland and French-speaking Switzerland. METHODS: The language of publication of 47,160 literature references cited in 2489 original articles published in the Swiss Medical Weekly between 1920 and 1995 was established. The 1730 articles published in German contained 32,607 assessable references; the 759 articles published in French contained 14,553 assessable references. RESULTS: The percentages of literature references in German, French and English cited in the articles written in German were, respectively, 83.6%, 9.1% and 5.9% in 1920; 68.6%, 7.2% and 18.3% in 1945; 30.7%, 5.6% and 61.9% in 1970; 11.3%, 1.5% and 86.7% in 1995. The percentages of literature references in French, German and English cited in the articles written in French were, respectively, 61.1%, 31.8% and 4.0% in 1920; 30.6%, 39.3% and 26.5% in 1945; 19.8%, 9.6% and 69.7% in 1970; 7.4%, 2.4% and 90.0% in 1995. CONCLUSIONS: (1) Between 1945 and 1995 the percentage of literature references in English has increased continuously, while the percentages of references in German and French have decreased. (2) English replaced German as the main language of medicine towards 1955 in French-speaking Switzerland and towards 1965 in German-speaking Switzerland. (3) During the whole period studied (1920-1995), French-speaking authors cited publications in German more frequently than German-speaking authors cited publications in French. (4) The evolution of the relative importance of the languages in German-speaking Switzerland is very similar to that previously described in Germany and Austria. (5) In French-speaking Switzerland, on the other hand, the evolution of the relative importance of the different languages differs very considerably from that previously described in France. PMID- 9411714 TI - [Maturity-onset diabetes of the young (MODY)--a heterogeneous disease picture]. AB - A family with maturity-onset diabetes of the young (MODY), a rare subtype of non insulin-dependent diabetes mellitus (NIDDM), is described. This familial form of diabetes is characterized by an early age of onset and an autosomal dominant inheritance. All three patients developed diabetes at an early age and were not obese. The diabetes was always well controlled and none of the patients developed late complications. The clinical and molecular-genetic characteristics of the different forms of MODY are described, as well as the diagnostic work-up and therapeutic management. PMID- 9411716 TI - [New drugs in the treatment of alcoholism]. AB - Pharmacotherapy of alcoholism is improving rapidly with the introduction of new agents. New knowledge about the neurobiology of alcoholism is necessary for the clinician, who has to establish the diagnosis. Useful pharmacological agents for the treatment of alcohol dependence can be classified into four groups: (1) agents for the treatment of the withdrawal syndrome, (2) aversive agents, (3) therapeutic agents for comorbidity, (4) new agents to reduce craving for alcohol or prevent relapse. These new agents derive from research in four directions, based on neurobiological hypotheses: (a) the glutamatergic hypothesis with acamprosate, (b) the opioid hypothesis with naltrexone, (c) the serotonergic hypothesis with the new antidepressants, and (d) other hypotheses, including dopaminergic, peptidic etc. Of these new agents, acamprosate has undergone most study in controlled clinical trials around Europe. Its efficacy has been demonstrated statistically, it is well tolerated and does not interact with alcohol. Acamprosate can be associated with disulfiram therapy. Future perspectives for treatment and research are discussed, in particular with regard to therapeutic associations. PMID- 9411717 TI - [Cutaneous artefacts]. PMID- 9411718 TI - [Tropical medicine: status yesterday and today]. PMID- 9411719 TI - [Risks of inline skating]. PMID- 9411720 TI - [The acquired immunodeficiency syndrome--l5 years on]. PMID- 9411721 TI - [The importance of HIV-1 in dentistry, oral medicine and orthodontics. A virological, epidemiological and clinical update]. PMID- 9411722 TI - [Emergency care in the dental office. Continuing education of the Ecole de Medecine dentaire (Geneva): the emergency course 101 of 20 June 1997]. PMID- 9411723 TI - [Homes: systematic care care desired]. PMID- 9411724 TI - [Written tests in "baseldytsch" (Basel German). Interview by Vivianne Berg]. PMID- 9411725 TI - [Plaque removal or plaque control. The 4th IHCF Annual Congress of 4 and 5 April 1997 in Leipzig. International Health Care Foundation]. PMID- 9411726 TI - [Sudden death of Alpine cattle in the canton Graubunden]. AB - The aetiology of sudden deaths of cattle in the Kanton Graubunden has been elucidated by a multi-disciplinary approach. Certain small rivers and ponds located in the affected Alpine pastures in the areas of Misox, Rheinwald and Engadin provide favourable habitats for the growth of hepatotoxic cyanobacteria, particularly during long periods of hot and dry weather. As cattle frequently take up water from these sources, the toxins produced by cyanobacteria may lead to lethal poisonings that are typically associated with hemorrhagic liver necrosis. The latest reported case of cyanobacteria poisoning of Alpine cattle occurred during the summer of 1996. Further animal losses may be avoided by the implementation of appropriate pasture management schemes. To identify sites of potential hazards to human or animal health, we have developed a sensitive screening assay for the detection of cyanobacteria hepatotoxins in algae and water samples. PMID- 9411727 TI - [Surgical treatment of carpal hygroma in cattle: 17 cases (1990-1994)]. AB - This retrospective study describes the case reports of 17 cattle suffering from precarpal hygroma, admitted to the clinic for food animals and horses, University of Berne, between 1990 and 1994. The following criteria were evaluated: sex, age, and bodyweight, case history, clinical findings at admission, surgical technique, aftercare, and short- and long-term results. The hygroma was congenital in 3 cases and acquired at the age of 2 to 84 months in 14 cases. Characteristic clinical findings at admission were a non painful, fluctuating, well delineated swelling of the precarpal area with a maximal diameter of 6 to 20 cm, which did not provoke any lameness. Treatment consisted of surgical excision of the bursa. Surgery was performed under general anesthesia with the animal in lateral recumbency and the affected limb positioned uppermost. After placing a tourniquet proximal to the carpal joint, routine preparation and draping of the surgical field, the bursa was resected completely, a penrose drain introduced, and primary wound closure attempted. Aftercare consisted of parenteral antimicrobial treatment and immobilisation of the affected limb with a full-limb splint bandage to prevent wound dehiscence and seroma formation. Primary wound healing was achieved in all cases. At the time of long-term follow-up evaluation, 4 to 48 months after surgery, telephone conversation with the owners revealed uncomplicated healing in 16 cases. In one case, recurrence of the hygroma had occurred a few weeks after surgical excision had been performed. PMID- 9411729 TI - [Attack of black flies (Diptera, Simuliidae) on horses in Basel (Switzerland)]. AB - In April 1996 an attack of black flies (Diptera, Simuliidae) to horses was observed in the region of Basle, Switzerland. Females of Simulium erythrocephalum and Simulium ornatum were involved in this attack, part of them had taken a blood meal. For S. ornatum this is the first record in Switzerland that this species takes blood meals on horses. PMID- 9411728 TI - [Hemorrhagic pericardial effusion in dogs. A retrospective study of 10 cases (1989-1994) with a review of the literature]. AB - After an introduction on pathophysiological and etiological aspects of idiopathic hemorrhagic pericardial effusion, ten dogs affected with this condition are described. Clinical signs permitted to establish a diagnosis of suspicion in all cases. Electrocardiography, thoracic radiography and echocardiography allowed confirmation of the diagnosis. Echocardiography was the most sensitive method available. Six dogs had idiopathic pericardial effusion. A subtotal pericardectomy was performed in two of these dogs. In four dogs, the effusion was caused by a tumor. Two dogs with mesothelioma were euthanized because of poor prognosis. Heart base tumors were found in the two other dogs. One of them is still alive and well one year after subtotal pericardectomy. The other one was euthanized due to deterioration of his condition two years after the operation. Idiopathic hemorrhagic pericardial effusion carries a good prognosis if appropriately treated (pericardiocentesis, possibly subtotal pericardectomy). Pericardial effusion is also frequently associated with tumors. In such cases, prognosis and therapy essentially depend on the type of tumor. PMID- 9411730 TI - [Revision of the animal welfare regulations]. PMID- 9411731 TI - [Clinical experiences with the therapy for patella luxation of small animals using sulcoplasty and lateral and cranial relocation of the tuberositas tibiae]. AB - Twenty cases with patellar luxation in dogs and cats, in which a sulcoplasty and cranialisation of the tuberositas tibiae were performed, were investigated and reviewed retrospectively. Twelve knees were available for follow-up after a mean period of 15 months. Clinical scoring of patients showed eight with no lameness, three with an occasional weightbearing lameness, and one with a frequent weightbearing lameness. The patella was stable and could not be luxated in ten out of twelve cases. Degenerative joint disease was slightly progressive in the postoperative period. It did not impair the outcome of the procedure. PMID- 9411733 TI - [Urinary incontinence in castrated bitches. Part 1: Significance, clinical aspects and etiopathogenesis]. AB - Acquired urinary incontinence occurs in 20% of spayed dogs and there exists a strong correlation between body weight and the risk of urinary incontinence. Bitches with a body weight of more than 20 kg have a risk of 30% white smaller dogs have a risk of 10%. A particular breed disposition exists in Boxers in which 65% are affected. Other breeds with a more than average disposition for urinary incontinence are Dobermans and Giant Schnauzers. Urinary incontinence due to spaying manifests itself mainly while the dogs are sleeping. The cause is a urethral sphincter incompetence which can be verified by a urethral pressure profile (UPP). The microtransducer method proved to be a suitable method for urodynamic studies. It could be demonstrated that the urethral closure pressure is significantly lower in incontinent bitches (4.6 +/- 2.3 cm H2O) than in continent bitches (18.6 +/- 10.5 cm H2O). In addition, the urethral closure pressure for continent bitches dropped significantly within 12 months after surgery. Histological examination revealed that the functional urethral closure cannot be explained by the extent of discernible structures of the urethral wall as seen by light microscopy. PMID- 9411732 TI - [The biomechanics of the hip joint using new diagnostic aspects in the field of hip dysplasia. Constructive critical thoughts on hip dysplasia diagnosis and today's marketable breeding methods with an outlook on future perspectives and possibilities. Part II]. PMID- 9411734 TI - [Case presentation: bovine leukocyte adhesion deficiency (BLAD) in Switzerland]. AB - Bovine Leukocyte Adhesion Deficiency (BLAD) Syndrome is a lethal congenital immunodeficiency caused by the strong reduction in the expression of leukocyte integrins (beta 2 integrins) on the surface of leukocytes. Therefore, neutrophils from BLAD animals lack the capacity to adhere to the endothelium, a necessary step in their emigration into foci of infection. Due to the virtual absence of neutrophil-mediated host defense, animals suffer from recurrent infection of the respiratory and gastrointestinal tracts and finally succumb to infections. A 14 days old Holstein-Friesian calf showing omphalophlebitis and leukocytosis, was referred to our clinic. It was found to suffer from several febrile episodes of infection. The tentative diagnosis BLAD could be confirmed for the first time in Switzerland by flow cytometry, pedigree analysis and by restriction fragment length polymorphism. PMID- 9411735 TI - [Cerebrospinal nematodiasis in seven goats]. AB - Clinical findings in seven goats affected with cerebrospinal nematodiasis are described. The animals originated from different parts of Switzerland. The disease occurred mainly in winter. The animals were admitted to the clinic because of progressive pelvic limb ataxia, recumbency, vestibular disease and circling. Clinical findings were complete or incomplete posterior paresis, pelvic limb ataxia, circling, reduced cutaneous sensation and proprioceptive deficits as well as cranial nerve reflexes deficits. The general condition was slightly reduced and the appetite was normal. In three goats predominance of mononuclear and eosinophilic cells in the cerebrospinal fluid was interpreted as typical findings for parasite infestation in the central nervous system. Histopathological changes and the finding of a nematode in cross sections in two affected animals confirmed the diagnosis. Infection with Elaphostrongylus cervi is discussed due to close contact with deer. PMID- 9411736 TI - [The therapy of equine sarcoid with a non-specific immunostimulator--the epidemiology and spontaneous regression of sarcoids]. AB - 20 sarcoid-affected horses from a practice in the northern Jura were used in this experiment. The mean age of the 20 horses was 3.9 years at the time of the first observation of sarcoid tumors. On the average, 4.4 tumours were noted per horse. 10 of the horses were treated in a double-blind study with an unspecific immunostimulant (Baypamun P), 10 others received a placebo. One single tumour only was treated per horse. The injections were given under and around the sarcoid. In eight out of the 20 horses all tumours regressed totally or for more than 50% of their initial size. Five of these had received placebo, three the immunostimulant. Four animals showed a modest, but measurable reduction in tumour size (3 immuno-stimulant, 1 placebo) and in the remaining eight horses (4:4) no reduction or even an increase in tumour size was observed. The phenomenon of tumour regression was very probably due to spontaneous regression and horses which had been observed to develop sarcoid within the last three months had significantly more regressions than animals with older tumours (p < 0.05). The haplotype of the equine leucocyte antigens was thought to predispose 12 of the 20 horses for the sarcoid. However, the ELA-type did not measurably influence the phenomenon of tumour regression. PMID- 9411737 TI - [Occurrence of bovine ehrlichiosis in the canton Obwalden]. AB - Due the examination of six Brown Swiss cows the "Schinberg-fever" a clinical picture known for years in central Switzerland could be identified as bovine ehrlichiosis. Clinical signs were observed 23 to 116 days after the animals were turned out on pasture. Main symptoms were fever and reduced milk yield. All animals were infested with ticks and showed a severe leucopenia. In buffy coat preparations Ehrlichia phagocytophila could be identified in neutrophils and eosinophils, rarely in monocytes. All six cows could be successfully treated with oxytetracycline. PMID- 9411738 TI - [Occurrence of Mycobacterium genavense in birds]. AB - A total of 253 birds were investigated to determine the presence of mycobacteria. Scrapings from various internal organs were stained according to Ziehl-Neelsen, and acid-fast bacilli were found in 26 birds (10.2%). Cultivation of mycobacteria was attempted from 22 livers, 12 spleens, 14 kidneys, 12 lungs, and 9 intestines from these 26 birds. Each sample was first decontaminated using a modified sodium dodecyl sulfate method, and three media were inoculated (Bactec 12 B; Lowenstein Jensen, Herrold). Using a Polymerase-Chain-Reaction-Restriction-Enzyme-Analysis (PRA), M. genavense could be identified in 19 of 26 birds (73%). M. avium could be isolated from three birds and M. fortuitum from one bird. In total, mycobacteriosis was the primary diagnosis made in 24 of 26 birds (92%). A presumptive diagnosis of mycobacteriosis was already made macroscopically in 14 of these birds. In the remaining 10 cases, bacteriological and histological investigations with specific staining methods were necessary for diagnosis. Several different histological changes were found. We observed individual macrophages as well as epitheloid cell proliferation, with variable, relatively mild inflammatory and fibrous reactions. We could not find any correlation between infection with a mycobacterium species and a specific tissue reaction pattern. This report demonstrates that M. genavense is an important cause of avian mycobacteriosis especially in pet birds. PMID- 9411739 TI - Gene technology and democracy. PMID- 9411740 TI - Affection for the MBL. PMID- 9411741 TI - Affection for the MBL. PMID- 9411742 TI - Affection for the MBL. PMID- 9411743 TI - Genetics of Parkinson's disease. PMID- 9411744 TI - Last-minute deal give NIH 7.1% raise. PMID- 9411745 TI - Editors seek ways to cope with fraud. PMID- 9411746 TI - An antidote to bioproliferation. PMID- 9411747 TI - The architecture of hearing. PMID- 9411748 TI - Molecules give new insights into deadliest brain cancers. PMID- 9411749 TI - HIV survives drug onslaught by hiding out in T cells. PMID- 9411750 TI - Receptor offers clues to how 'good' cholesterol works. PMID- 9411751 TI - The early Mars climate question heats up. PMID- 9411752 TI - A Myc-induced apoptosis pathway surfaces. PMID- 9411753 TI - The civil commitment of sex offenders. PMID- 9411754 TI - Immune response and myoblasts that express Fas ligand. PMID- 9411755 TI - Twin studies, heritability, and intelligence. PMID- 9411756 TI - Twin studies, heritability, and intelligence. PMID- 9411757 TI - Twin studies, heritability, and intelligence. PMID- 9411758 TI - Twin studies, heritability, and intelligence. PMID- 9411759 TI - Consensus on African research projects. PMID- 9411760 TI - A womb with a view. PMID- 9411761 TI - How does HIV overcome the body's T cell bodyguards? PMID- 9411762 TI - Will fossil from down under upend mammal evolution? PMID- 9411763 TI - Rat model for Gulf War syndrome? PMID- 9411764 TI - Protective role for prion protein? PMID- 9411765 TI - Heat shock protein linked to stroke protection. PMID- 9411767 TI - Laser capture microdissection: molecular analysis of tissue. PMID- 9411766 TI - Methane: small molecule, big impact. PMID- 9411768 TI - Using one's head. PMID- 9411769 TI - Dopamine's role. PMID- 9411770 TI - Dopamine role. PMID- 9411771 TI - Olfactory "consciousness"? PMID- 9411772 TI - Sexual selection in Asian elephants. PMID- 9411774 TI - A 5-year initiative slowly takes shape. PMID- 9411773 TI - Bangkok study adds fuel to AIDS ethics debate. PMID- 9411775 TI - Multiple clocks keep time in fruit fly tissues. PMID- 9411776 TI - Sex frees viruses from genetic 'ratchet'. PMID- 9411777 TI - Viruses scout evolution's path. PMID- 9411778 TI - New developmental clock discovered. PMID- 9411779 TI - The rise and fall of Project SIDA. PMID- 9411780 TI - Fashioning flow by self-assembly. PMID- 9411781 TI - A new player in cell death. PMID- 9411782 TI - Variations on a theme: cataloging human DNA sequence variation. PMID- 9411783 TI - "Gene war of the century"? PMID- 9411784 TI - The relaxation response: therapeutic effect. PMID- 9411785 TI - Japan centers in on genome work. PMID- 9411786 TI - Drug companies decline to cooperate. PMID- 9411788 TI - Putative Martian microbes called microscopy artifacts. PMID- 9411787 TI - Spiked coffee prompts police inquiry. PMID- 9411789 TI - Herpesvirus linked to multiple sclerosis. PMID- 9411790 TI - Salvation in a snippet of DNA? PMID- 9411791 TI - World AIDS--the worst is still to come. PMID- 9411792 TI - The ins and outs of protein translocation. PMID- 9411793 TI - Pharmacia Biotech & Science prize. 1997 grand prize winner. Life in the balance: cell walls and antibiotic resistance. PMID- 9411794 TI - Mars Pathfinder [foldout]. AB - The following foldout present images and analysis from the Mars Pathfinder Mission that are discussed in seven subsequent Reports. The center is a four-page panorama of the surface of Mars around the lander (Plate 1). The back of the foldout contains surface images (Plate 7), a different perspective of the landing site (Plate 2), rover targets (Plate 3), locations of rocks and other features (Plate 6) and data analysis (Plates 4, 4, 8, 9, and 10). PMID- 9411796 TI - [Initial cranial CT for evaluating the prognosis of craniocerebral trauma]. AB - A total of 208 multiple trauma patients with head injury (HI) were investigated who had been treated in the period from 1990 to 1995. The average age was 35.2 +/ 17.7 years; the injury severity according to ISS was 30.2 +/- 8.6 points; 20.5% died as a result of the HI; the mortality of all patients was 26.5%. The Glasgow Coma Scale (GCS) was determined at an average of 22 min after trauma (8.0 +/- 4.3 points) at the scene of accident. The patients were classified according to GCS into minor HI (group 1: 14-15 points), moderate HI (group 2: 9-13 points) and severe HI (group 3: 3-8 points). Patient outcome was assessed by the Glasgow Outcome Scale (GOS) and was classified as good (GOS 4 and 5) and poor (GOS 1, 2 and 3) outcome. At the latest, 2 h after trauma, a CT scan of the head (CCT) was done. The HI groups are compared regarding frequency of types of injury. In all HI groups the fractures of the bony face occurred at the same frequency (36.0 38.9%). The frequency of calotte fractures (Kal-Fx) increased from group 1 (8.0%) to 2 (19.2%) and 3 (25.6%); fractures of the skull base significantly differed between group 1 (16.0%), 2 (7.8%) and 3 (33.4%). Epidural hemorrhage (EDB) appeared only in group 2 (7.8%) and 3 (6.7); subdural hemorrhage was found in group 1 (2.7%), 2 (7.8%) and 3 (10.0%). Subarachnoid hemorrhage (SAB) was significantly more frequently seen, dependent on HI severity, in group 3 (26.7%) compared to group 2 (11.7%) and 1 (8.0%). Intracerebral contusion (ICK) significantly increased from group 1 (12.0%) to 2 (27.3) and 3 (45.6%). Brain swelling (BS) also significantly increased from group 1 (8.0%) to 2 (19.5%) and 3 (49.0%) and lesions of ventricles (VL) from group 1 (2.7%) to 2 (11.7%) and 3 (20.0%). Midline shift (13.4%) and signs of herniation (4.5%) only occurred in group 3. The analysis of correlation/regression and receiver operating characteristics was able to predict 79% of patients' outcome accurately using GCS (r 0.54; P < 0.0001) alone, using CCT (r 0.65; P < 0.0001) 87% were correctly predicted with significant variables Cal-Fx, EDB, SAB and BS. CCT with GCS (r 0.74; P < 0.0001) were able to predict 88% accurately with significant variables Cal-Fx, EDB, BS and GCS. The combination of CCT with GCS, age and ISS (r 0.78; P < 0.0001) was able to predict only 87% correctly, although the r value was the highest; significant variable were Kal-Fx, EDB, BS, VL, GCS, age and ISS. PMID- 9411795 TI - [Surgical replantation]. AB - In the early days of replantation surgery, if viability was restored the operation was judged a success. Nowadays restoration of viability alone is not sufficient to fulfill the criteria of successful replantation, which are as follows: Lack of severe systemic disturbances due to the replantation, a "functional extremity" according to the definition of Chen et al. (1978), no or little pain at the site of the replantation, good aesthetic results, and an acceptable length of time for rehabilitation and return to normal life. Successful replantation needs a therapy concept that is based on an exact definition of the amputation injury from the viewpoint of the amount of severance, the level of the amputation, and the type of amputation mechanism, complete knowledge of current replantation indications, and exact selection of patients amenable for replantation. PMID- 9411797 TI - [Enchondroma of the hand. Clinical evaluation study of diagnosis, surgery and functional outcome]. AB - Enchondroma are benign cartilaginous tumors and are localized most often at the site of the phalanges. Between 1982 and 1993 73 patients with monostotic enchondroma and 5 patients with polyostotic enchondroma were operated at our clinic. Clinical signs of monostotic tumors were pathological fracture (38.4%), pain or swelling. Eleven percent of cases were accidental findings. Surgical treatment was performed by complete removal of the tumors and filling the bone cavity with autologous spongiosa taken from the pelvic bones, the elbow, or the radius. Three patients (4.1%) had to be operated a second time due to wound infections and hematoma. In one case Sudeck's dystrophy was diagnosed. One patient (1.4%) developed a recurrent tumor. Our follow-up examination of 65 patients showed that 77% of the patients with monostotic enchondroma achieve very good or good functional long-term results after this operation, but only 40% of the patients with polyostotic enchondroma. PMID- 9411798 TI - [Unreamed intramedullary nailing as minimal invasive palliative intervention in osteolysis and pathologic fractures of long tubular bones]. AB - Pathological fractures and osteolyses with impending fractures of long bones impose a severe problem in patients with advanced cancer. For palliative treatment between June 1993 and October 1995 intramedullary nailing was performed in 19 patients with 13 femoral, 7 humeral and 1 tibial lesions; AO unreamed nails were used for this purpose (unreamed femur nail UFN with or with spiral blade for femoral lesions and unreamed tibia nail UTN for humeri and tibia). Stabilization was achieved without severe complications; full weight bearing of both femora and humeri (while using crutches) was allowed. Simultaneous and bilateral nailing was done. Follow-up until death due to advanced cancer demonstrated no implant failure and improved quality of life as well. Unreamed intramedullary nailing seems to be a minimally invasive procedure with a low complication rate for palliative treatment of (impending) pathological fractures in advanced cancer. PMID- 9411799 TI - [Proprioceptive capacities of patients with retropatellar knee pain with special reference to effectiveness of an elastic knee bandage]. AB - In the presented study, knee joint proprioception of 43 patients with a patellar pain syndrome of the knee joint was evaluated. In a control group, the proprioception of 30 healthy volunteers with clinical and an-amnestic inconspicous knee joints was examined. We tested the proprioceptive capability of the subjects with a passive angle reproduction test. Additionally, all knee joints were measured with and without an elastic knee bandage. The patient group showed significant deterioration of angle reproduction capability (13.2 degrees +/- 6.1 degrees) compared to the control group (7.8 degrees +/- 2.8 degrees). After applying an elastic knee bandage, the angle reproduction capability significantly improved to 9.2 degrees +/- 4.5 degrees. Proprioception of the contralateral, noninvolved knee joint in the patients (11.6 degrees +/- 6.3 degrees) was worse compared to the control group. Applying an elastic knee bandage did not significantly improve the proprioception of the uninjured knee joint. PMID- 9411800 TI - [Replacement of the anterior cruciate ligament by cold preserved bone-cruciate ligament-bone allotransplants. An experimental study in the sheep]. AB - One ACL in each of 17 mature sheep was replaced with a deep-frozen bone-an ACL bone allograft. Allografts were obtained from skeletally mature sheep using a standard aseptic technique and stored deep frozen for at least 6 days (mean 21 days). Macroscopical, biomechanical, and histological changes were evaluated 12, 24, and 52 weeks following implantation. At autopsy all allograft ligaments were present and demonstrated no evidence of infection or immune reaction. We found slight arthrotic changes in 3 knees after 12 weeks, in 4 knees after 24 weeks, and in 3 knees after 52 weeks. Twelve weeks after the operation the maximum load of the allografts was 17.5% of the contralateral controls and increased to 20.9% after 24 weeks and to 32% of controls after 52 weeks. Ligament stiffness in the linear region also increased from 18.9% of control (12 weeks) to 32.5% after 52 weeks, whereas maximum load decreased from 112.2% of controls (12 weeks) to 98% of controls (52 weeks). Histologically, the allografts progressively matured with time, becoming nearly identical to normal ligaments at 52 weeks. PMID- 9411801 TI - [The Chopart dislocation. A frequently underestimated injury and its sequelae. A clinical study]. AB - During a 7-year period (between July 1987 and June 1994) 19 patients were treated for Chopart dislocation. Seventeen patients were examined clinically 48 months following trauma. We found 1 excellent, 6 good, 6 mediocre, and 4 poor results. Only 4 patients were able to achieve their pre-accident level of work and no patients could return to sports at the same level. In our patients, Chopart dislocation had a negative social consequence both at the work place and in recreational activities. In long-term follow-up, the Chopart dislocation was the most debilitating injury even for polytrauma patients. PMID- 9411803 TI - [Mobilization of the tibial insertion of the anterior cruciate ligament: new therapeutic possibilities in surgery of the cruciate ligament]. AB - In patients with a significant elongation of the ACL we propose a new therapeutic regimen. After mobilisation of the tibial insertion of the ACL we pull the bone block with the attached ACL distally. Thus the ACL is easily tightened. In accordance with this method, we describe the treatment of a case of a strictly proximal rupture of the ACL: the tibial insertion is mobilised, the bone block with the attached ACL removed from the joint. Ex vivo sutures are placed into the ACL, the ligament is pulled back into the joint, and the shortening of the ligament caused by the sutures is compensated by slight proximal dislocation of the bone block. PMID- 9411802 TI - [Fractures of the tibial plateau]. PMID- 9411804 TI - [Prevention of thrombosis in trauma surgery--current legal aspects]. AB - To date, current jurisdiction has not permitted an answer to the a question as to how long thromboembolism prevention therapy is to be carried out after trauma surgery. However, the Bundesgerichtshof (BGH) reached a decision in 1995 on this matter. Accordingly, patients have to be informed about possible alternative treatment as well as the applicable form of thromboembolism prevention therapy in each specific case. In combination with the fact that the Deutsche Gesellschaft fur Phlebologie (DGP, German society for phlebology) has published guidelines on thromboembolism prevention therapy via the internet, this may have considerable impact on the outcome of civil litigation, with consequences for financial compensation. Since civil action requires the medical doctor to prove that he has provided an adequate and acceptable basis for informed consent in patients, in concurrence with the above-mentioned DGP guidelines being potentially misunderstood as representing state-of-the-art treatment, it is deemed necessary to present a list of points that must be brought to the patient's attention when requesting informed consent and, furthermore, the manner in which the guidelines of the medical specialist societies may bring about problems must be pointed out. In the specific context of the German judicial system, criminal law will virtually never lead to a conviction in a case of fatal pulmonary embolism following neglect of or insufficient thromboembolism prevention therapy. PMID- 9411805 TI - The Fox Chase Cancer Center and Free University Hospital Investigators' Workshop and Consensus Conference on Paclitaxel. Part 3: Breast Cancer. Puerto Rico, March 12-16, 1997. Proceedings. PMID- 9411806 TI - [Technetium 99m tetrofosmin as a myocardial perfusion marker in diagnosis of coronary heart disease. Validation a one day rest-stress protocol]. PMID- 9411807 TI - [Extravasation of contrast media into the pericardium--a rare complication of intravenous digital subtraction angiography?]. PMID- 9411808 TI - [Women in radiology]. PMID- 9411809 TI - [Costs for after care in breast carcinoma]. PMID- 9411810 TI - Communication for health professionals in a multilingual society. PMID- 9411811 TI - Education of health professionals for a restructured health system - whose responsibility should it be? PMID- 9411813 TI - Halitosis. PMID- 9411812 TI - Footprints: Anthony and Margaret Barker. PMID- 9411814 TI - Bunji jumping discouraged! PMID- 9411815 TI - Epidemic Shigella dysentery in South Africa. PMID- 9411816 TI - [Problem Falck--11,600 ambulances arrive only after 15 minutes]. PMID- 9411817 TI - [Problem Falck--false security]. PMID- 9411818 TI - [The treatment of arterial hypertension]. PMID- 9411819 TI - [The polymorphism of the angiotensin-converting enzyme gene in patients with hypertension, left ventricular hypertrophy and the development of a myocardial infarct at a young age. Preliminary report]. AB - Insertion/deletion (I/D) polymorphism of angiotensin-I-converting enzyme (ACE) gene was studied by use of the polymerase chain reaction in 168 normal subjects living in Moscow region and in 70 patients: 38 with essential hypertension (EH), 9 of which survived myocardial infarction at young age, 13 with hypertrophic cardiomyopathy (HCMP) and 19 with myocardial infarction (MI). Left ventricular hypertrophy (LVH) was detected in 24 of 38 EH patients. There was a highly significant increase in the frequency of the ID genotype in EH patients compared to the controls (62.4% versus 32.7%, P < 0.01). There was a relevant decrease in the frequency of the DD genotype in EH patients in comparison with the control (20.8% versus 47.6%, P < 0.05). These results strongly suggest that the ACE gene is associated with EH. No significant differences in both allele and genotype frequencies of the ACE gene were revealed in two groups of patients with MI and with HCMP compared to the controls. Thus, no relations between the ACE gene and these disorders were observed. In hypertensives with MI the II genotype was not detected and the frequency of both D allele and DD genotype was sufficiently increased compared to normotensive patients with MI. Thus, the DD genotype in hypertensives may be a risk factor for MI. The frequency of the DD genotype was significantly increased in hypertensive patients with LVH compared to the uncomplicated hypertension (37.5% versus 7.1%, P < 0.05). Therefore, this genotype is associated with LVH in hypertensive subjects. PMID- 9411820 TI - [The characteristics of catecholamine metabolism in patients in the initial stage of hypertension]. AB - Catecholamines metabolism (24-h excretion, circadian rhythm of DOPA, dopamine, homovanillic acid, adrenaline, noradrenaline, vanilmandelic acid) was investigated in 124 young males with stage I essential hypertension treated by cranial electrostimulation. Relative activity of this metabolism stages was assessed. Patients with early stages of essential hypertension seem to have enhanced secretory and metabolic activity of the sympathetic link of the sympathoadrenal system with associated dopamine dysbolism; relative deficiency of dopaminergic system with activation of dopamine conversion to homovanillic acid in its intact conjugation. It is suggested that a marked increase of the ratio noradrenaline/dopamine may serve a biochemical marker of essential hypertension and risk of the disease progression. PMID- 9411821 TI - [Magnetic resonance tomography and magnetic resonance phlebography in studies of the brain in patients with severe arterial hypertension]. AB - MR tomography of the brain, MR angiography of extra- and intracranial arteries and veins, duplex scanning of the extracranial arteries and veins were performed according to the technique spin-echo, by the technique 2D, 3D time-to-flight (unit "Magnetom 63 SP", 1.5 T), Acuson 128 technique, respectively, in 21 patients with high and malignant blood hypertension (BH) and 11 healthy controls. In BH patients magnetic resonance has detected signs of hypertensive encephalopathy: dilation of the liquor-conduction system, small hyperintensive foci in the white substance, zones of periventricular hyperintensity. MR angiograms visualized flexures of the extracranial arteries. MR phlebograms were characterized: 1) reduction of or absence of the signal from the blood flow along the cross, sigmoid sinus and internal jugular vein of the hemisphere associated with enlargement of the contralateral structures; 2) lowering of the signal intensity from the blood flow along the upper sagittal sinus; 3) dilation of the emission veins and superficial cerebral veins. These abnormalities should be considered in the treatment of BH. PMID- 9411822 TI - [Lipoprotein(a) as a biochemical marker of coronary atherosclerosis]. AB - The relation between lipoprotein Lp(a) levels, other lipids and severity of coronary atherosclerosis was studied in 281 patients (241 men and 40 women aged 24 to 68 years) with suspected or diagnosed coronary heart disease. All of them underwent coronary angiography. The angiograms were evaluated according to two scores: vessel score (0 to 3 points for 0 to 3 vessels with stenoses > 50%) and stenosis score (0 to 32 points, number and severity of coronary lesions). 224 patients with verified coronary atherosclerosis were grouped according to these scores and compared with subgroup of 57 patients with intact coronary arteries. Lp(a) levels were significantly higher in patients with 1-, 2- (p < 0.05) and 3 vessel (p < 0.01) disease and stenosis score more than 10 points (p < 0.01). LDL cholesterol (LDL-C) levels were significantly higher in patients with 2- and 3 vessel disease and stenosis score more than 10 points (p < 0.05 in all cases). There was a positive correlation between Lp(a) levels and severity of coronary atherosclerosis (p < 0.05). Lp(a) contributes < 15% of total plasma cholesterol (TC). After subtraction of Lp(a)-cholesterol from TC according to modified Friedewald formula there was no any correlation between Lp(a) and LDL-C levels. PMID- 9411823 TI - [The antioxidant probucol as a regulator of the intensity of free-radical lipid peroxidation processes in the blood of patients with coronary atherosclerosis]. AB - Two series of studies were made to assess probucol medicines (lipomal, "Alkaloid"; fenbutol "Akrikhin") effect on clinical symptoms, lipid metabolism, primary and secondary products of lipid peroxidation, activity of antioxidant enzymes (superoxide dismutase, glutathione peroxidase) in atherosclerotic patients with hyperlipidemia type IIA and IIB. In both series probucol reduced frequency of anginal attacks, content of total cholesterol and LDL cholesterol. HDL cholesterol remained unchanged or reduced insignificantly. Lipoperoxides and malonic dialdehyde levels in plasma progressively lowered against activation of antioxidant enzymes. These biochemical parameters returned to initial values within 3 months since the drug discontinuation. It is evident that antiatherogenic effect of probucol is due to indirect activation of natural defense systems responsible for enzymic detoxication of active oxygen forms and lipoperoxides rather than to direct interaction of this synthetic antioxidant with lipid radicals. PMID- 9411824 TI - [The effect of long-term Enduracin monotherapy on the clinical and biochemical status of patients with ischemic heart disease]. AB - 13 patients aged 39 to 60 years with coronary atherosclerosis confirmed at selective coronary angiography combined with primary hyperlipidemia (phenotypes 2a and 2b) received enduracin in a dose 1500 mg/day. As a result of the treatment total cholesterol (TC) and LDL cholesterol lowered by 10.3 and 13.1%, respectively, whereas HDL cholesterol rose by 15.2%. Half of the patients demonstrated activation of hepatic transaminases, but discontinuation of the drug was not necessary. In 3 out of 4 patients after 2 years of enduracin treatment stabilization of atherosclerosis was observed. Thus, long-term enduracin is able to inhibit progression of atherosclerosis in coronary heart disease patients. PMID- 9411825 TI - [The effect on painful and silent myocardial ischemia of the preparations Efox and Efox Long in patients with stenocardia of effort]. AB - The study of two novel drugs for management of both painful and silent myocardial ischemia isosorbide-5-mononitrate (IS-5-MN), efox, and its long-acting variant efox-long demonstrated that both drugs exerted noticeable antianginal effect at maximal concentration in blood. This was achieved by a single dose and course administration. The differences in the effects of the two dosage forms of IS-5-MN on painless ischemia may be explained by limited coronary reserves of anginal patients. PMID- 9411826 TI - [The possibility of the development of isosorbide dinitrate and nifedipine withdrawal syndromes in patients with stable stenocardia of effort]. AB - Exercise tests were performed in 18 patients with stable angina of effort treated with isosorbide dinitrate (ID) or nifedipine (NF) to see whether sharp discontinuation of the above drugs may entail withdrawal syndrome. Exercise tolerance declined 21 and 24 hours after the last administration of NF, 13 and 18 hours after that of ID. Compared to NF, ID tolerance decreased in a less degree. Some of the patients experienced myocardial ischemia in the course of the exercise tolerance tests, there were more frequent anginal episodes, arterial pressure rose. For ID withdrawal syndrome manifested weaker than for NF. PMID- 9411827 TI - [Means for increasing treatment efficacy in patients with ischemic heart disease]. PMID- 9411828 TI - [The evaluation of the antianginal effect of Corvaton-Retard in patients with ischemic heart disease by using paired bicycle ergometry]. PMID- 9411829 TI - [The results of exposure to an antisclerotic vegetarian diet enriched with soy based products on patients in the secondary prevention of ischemic heart disease]. AB - The effects of an atherogenic vegetarian diet enriched by soya-based products were investigated for the first time in this country. Clinical status and biochemical parameters of 32 patients suffering from coronary heart disease were studied. Groups 1, 2 and 3 were on the diet for 11-17, 19-22 and 30-40 days, respectively. Hyperlipidemic medicines were discontinued. The vegetarian diet resulted in normalization of the serum lipid spectrum. The most pronounced effect was achieved in group III. The developed vegetarian diet neutralized the adverse effects (an increase of cholesterol and triglycerides) of beta-blockers. PMID- 9411830 TI - [A trial of the long-term use of Sedacorone (amiodarone) under polyclinic conditions for the prevention of paroxysms of atrial fibrillation]. AB - Sedacorone (amiodarone) was given to 70 outpatients with coronary heart disease for 1.5-2 years to prevent paroxysms of cardiac arrhythmia after recovery of sinus rhythm. Sedacorone was administered initially in the dose 600 mg/day for 10 12 days, then the dose lowering was adjusted to the tolerance, heart rate and ECG readings. The minimal dose of 200 mg/day was taken for 5 days a week with a 2-day interval. In adequate individual dose Sedacorone prevented paroxysms of cardiac fibrillation or made the paroxysms less frequent in 91.4% of patients. Careful selection of patients and regular control helped avoid serious cardial and extracardiac complications. Sedacorone is recommended as a first-line medicine for outpatients with frequent paroxysms of cardiac fibrillations after recovery of the sinus rhythm to prevent the paroxysms recurrences. PMID- 9411833 TI - [Thrombocyte IIb/IIIa receptor inhibitors--a new direction in antithrombotic therapy]. PMID- 9411834 TI - [The unwanted effects of antihypertensive agents of the basic groups (2)]. PMID- 9411832 TI - [The effect of long-term Fraxiparin treatment on hemostasis in patients with primary pulmonary hypertension]. AB - The aim of the study was elucidation of hemostatic effects of low-molecular heparin Flaxiparin in patients with primary pulmonary hypertension (PPH). 10 PPH patients (mean age 39.0 (+)- 3.2 years, mean history of the disease 5.1 (+)- 0.9 years) were treated up to 6 months. For the first month Flaxiparin was injected in therapeutic doses 15,000 AXa ICU, twice a day. The next 5 months prophylactic doses were administered twice a day (7,500 AXa ICU). D-dimer, fragment 1 + 2, complex thrombin-antithrombin, beta-thromboglobulin, protein C, antithrombin III, antigen of tissue plasminogen activator and inhibitor of tissue plasminogen activator of type I, activity of the latter were measured before the treatment, after the therapeutic and prophylactic courses, 6 months after the treatment. Initially, the patients had procoagulative hemostatic disorders. i.e. activation of blood coagulation; fibrinolytic system was also affected. In the course of Flaxiparin therapy blood coagulation and fibrinolysis improved significantly. However, the effect was not persistent after the drug discontinuation. Flaxiparin can be recommended for treatment of PPH. PMID- 9411835 TI - [The sleep apnea syndrome and arterial hypertension]. PMID- 9411831 TI - [The use of Cordanum in combination with Corinfar-Retard for the treatment of ectopic arrhythmias in the sick sinus syndrome in patients with ischemic heart disease]. AB - 38 patients with ischemic heart disease (IHD) and sick sinus syndrome (SSS) received combined therapy with nifedipine (Corinfar-Retard) and talinolol (Cordanum). The former drug had a positive chronotropic effect on the heart, the latter's chronotropic effect was slightly negative. All the patients had sinus bradycardia and ectopic arrhythmia which needed therapeutic correction: supraventricular and ventricular extrasystoles, fibrillation paroxysms or/and atrial flutter, paroxysmal supraventricular tachycardia, ventricular tachycardia. Cordanum was given in a dose 50 mg twice a day, Corinfar-Retard 20 mg twice a day for 16 days. 30 patients responded to the treatment. In addition to good subjective response, episodes of extrasystoles, paroxysms, flutter and fibrillation occurred much less frequently. Side effects resulted in the treatment discontinuation in 3 patients. PMID- 9411836 TI - [Endothelial function in arterial hypertension and other risk factors for the development of atherosclerosis (a review of the literature--2)]. PMID- 9411837 TI - [Hypertriglyceridemia as a risk factor for the development of atherosclerosis]. PMID- 9411838 TI - [Reliable when the approach determines the size of the intervention. Interview by Dr. Klaus Reinhardt]. PMID- 9411839 TI - [The history of laparoscopy]. AB - Laparoscopy has for a long time been the domain of some very few people. The first physicians to perform "coelioscopy" routinely where internists about 1920. Later, gynecologists where the ones to make the most important developments and inventions. The actual start of modern laparoscopy in surgery was the first video endoscopic cholecystectomy, performed by the french surgeon Mouret in 1987. From this time on, laparoscopy for diagnostics and cholecystectomy became a new standard within about just five years. This article summarizes the history of laparoscopy from the ancient time to the present. PMID- 9411840 TI - [Laparoscopic adrenalectomy]. AB - Minimally-invasive adrenalectomy is a new and attractive procedure, which has advanced to the method of choice in the surgical treatment of benign adrenal diseases. This technique is contraindicated in the treatment of adrenal malignancies. The endoscopic removal of large benign adrenal tumors (> 6-8 cm) may be difficult. The preoperative management of the patients does not differ from the preparation for conventional adrenalectomy. Like in open adrenalectomy several different endoscopic approaches to the adrenals have been described: the transperitoneal approach with the patient in supine position, the transperitoneal approach in lateral position, the retroperitoneal approach in lateral position and the retroperitoneal approach in prone position. Excellent results have been achieved with all four techniques. The approaches with the patient in lateral position have the disadvantage that in case of bilateral adrenalectomy the patient has to be repositioned during the operation. Large tumors are more difficult to be removed with the retroperitoneal techniques, because of the limited space in the retroperitoneum. Operative times and conversion rates have been markedly reduced with increasing experience with these techniques. PMID- 9411841 TI - [Status and outcome of laparoscopic appendectomy--results of a prospective study of 600 consecutive appendectomies]. AB - In a prospective study 600 consecutive appendectomies were held. In 51 cases the surgery was conventionally ended after laparoscopic beginning. The laparoscopic proceeding was set independently of the dimension of the inflammation. Also the perforation did not exhibit any contraindication. The rate of complications of 8.7% was in the range of the conventional appendectomy. We, however, could show that with increased experience and save technique the morbidity, especially the septic complications could have been sinked to 2%. The duration of hospitalisation amounted on an average of 5.0 days, the surgery time 53.6 minutes. The main advantage of the laparoscopic proceeding is the outstanding overview with the possibility of a diagnostics of the abdominal cavity. Besides an acute appendicitis of which in 14.6% a perforation was present, relevant side findings were elevated in 11.3% which in 7.7% were endoscopic surgically treated at the same time. PMID- 9411842 TI - [Indications, technique and outcome of laparoscopic splenectomy]. AB - Since introduction of stapling instruments, which allow a safe hemostasis of hilar spleen vessels, the advantages of laparoscopic procedures are as well available for the laparoscopic splenectomy, like atraumatic dissection, reduced hospital stay, less bleeding and therefore less necessity of blood transfusion, and reduced analgetic consume. Laparoscopic splenectomy causes a lower morbidity comparing to open conventional splenectomy. Frequent indications for elective splenectomy are immune thrombocytopenic purpura, spherocytosis, Non Hodgkin Lymphoma and Hodgkin's disease. Limiting factors to laparoscopic splenectomy may be the size of spleen, concomiting diseases as portal hypertension and adhesions in upper left abdomen. To remove a big sized spleen we placed it within a large specimen retrival bag, which was partly delivered through a port in the left flank to allow digital morcellation of the spleen and piecemal removal. Eight females and fifteen males with a median age of 45 years (16-73) underwent splenectomy of immune thrombocytopenic purpura (14), spherocytosis (2), Non Hodgkin Lymphoma (5), Hodgkin's disease (1) and hair cell leukemia (1) with a spleen weight of 985 grams. The median operating time was 94 min (40-165) and median blood loss was 214 ml (50-400). Accessory spleens were removed in two cases. The median postoperative hospital stay was 5.6 days (3-10). The only complication was a parechymatous bleeding which needed conversion to open splenectomy. With careful selection of patients, laparoscopic splenectomy can be safely performed on normal and even enlarged spleens. The lateral approach is safe and offers excellent visualization of the splenic vessels, pancreas, and accessory spleens. PMID- 9411843 TI - [Transoral video-endoscopic esophageal diverticulostomy--a new treatment method for Zenker's diverticulum]. AB - 70% of the esophageal diverticula are pharyngoesophageal diverticula. They are caused by a malfunction of the superior esophageal sphincter. Surgical treatment consisted of a myotomy of the superior esophageal sphincter and the resection of the diverticulum through a lateral cervical incision. With the described new endoscopic transoral technique the septum between the diverticulum and the esophagus is divided by a stapling device and both lumina are combined resulting in a esophago-diverticulostomy. At the same time a myotomy of the transverse part of cricopharyngeal muscle and the proximal esophageal wall is performed. Due to the design of the stapling device usually a small pouch remains at the bottom of the diverticulum. A modification of the stapler instrument allowed us to shorten the length of this pouch. The time of the endoscopic procedure is markedly reduced compared to the conventional surgical intervention. There is no necessity for special postoperative wound care and the hospital stay is considerably shortened. Furthermore serious complications like fistulas and lesions of the recurrent nerve, encountered with the open surgical intervention, are avoided. This new technique combines the advantages of a minimal invasive approach and the development of new instruments with the confided principles of the conventional operation in treating the pathogenetical mechanisms of Zenker's diverticulum. To assess the value of the technique long term follow up studies as well as comparative trials will be needed. PMID- 9411844 TI - [Surgical therapy of morbid obesity: indications, technique of laparoscopic gastric banding and initial results]. AB - Severe obesity with co-morbidity such as diabetes mellitus, cardiac failure, obesity hypoventilation, degenerative bone diseases and increased incidence of malignancy give rise to shorter life expectancy and have an impact on quality of life. This results in higher costs of health care and work absence. Surgical procedures have become commonplace in the therapy of morbid obesity because of the infrequent success of medical treatment. We performed a horizontal gastroplasty by laparoscopic adjustable silicon gastric banding (LASGB) on 60 patients between 1. 11. 1995 and 28. 2. 1997. The average excess above normal weight was 62 kg, the median BMI (Body-Mass-Index) was 46.44 kg/m2. Fifty-nine procedures were performed by the laparoscopic method and one with an open technique. The average postoperative hospital stay was five days. Due to dorsal slipping or pouch enlargement the procedure had to be repeated on 6 patients (10%). The median loss of weight in the first three months was 14.78 kg, after six months 24.14 kg and after nine months 35.1 kg. Insulin treatment for three patients suffering diabetes mellitus could be discontinued-in addition blood sugar levels in six patients normalised. Two patients with obstructive sleep apnea syndrome no longer needed a nocturnal Nasal-Continuous-Positive-Airway Pressure-(nCPAP-)Therapy. To provide a better quality of life to this group of patients, the gastric banding is a suitable method for carefully evaluated and followed patients. In addition improved ability to work and reduction of health care costs due to co-morbidity and joint diseases have a positive economic impact. PMID- 9411845 TI - [Thoracoscopic operations of the spine]. AB - Endoscopic procedures of the spine have been utilized in the recent past with increased frequency. It is possible to reach any level of the spine with the endoscope. Indications for spine surgery via thoracotomy is the same as for thoracoscopic approaches to the spine. There are clear advantages of the endoscopic technique over the open procedure. The cosmetic result is better, the hospital stay and the stay in the ICU is shorter, the blood less is less as is the need for pain medication. Patients return to work earlier. It seems that this procedure is safe and cost effective. The indications are: thoracic disc herniation, infection, tumor, fracture, and ventral release for scoliosis and kyphosis. PMID- 9411846 TI - [Video-assisted thoracoscopic surgery--indications, technique and results]. AB - Video-assisted thoracoscopic surgery became an important tool in the surgical treatment of various thoracic disease. Currently many interventions which routinely required thoracotomy can be performed by VATS safely and with excellent results. This includes pleurectomy, decortication, wedge-resection, bullectomy and volume reduction surgery for emphysema, biopsy and/or resection of mediastinal tumors, thymectomy for myasthenia gravis, sympathectomy and even lobectomy. The benefit of thoracoscopic surgery is reduced postoperative pain, including diminished impairment of pulmonary function, shorter hospital stay and the more rapid recovery. PMID- 9411847 TI - [Minimal invasive surgery in breast carcinoma]. AB - The presence or absence of involved axillary lymph nodes is the single best predictor of survival of breast cancer, and important treatment decisions are based on it. For staging purpose as well as for local control a level I and II dissection is recommended. In order to lower the morbidity of axillary lymph node dissection less invasive treatment modalities have been evaluated. Beside the sentinel lymph node biopsy another new method is discussed: the endoscopic axillary lymph node dissection. After liposuction of the axillary fat the lymph nodes of level I and II are removed by endoscopy. With this technique enough lymph node can be removed which allows a sufficient staging as well as local control. This technique is not to be recommended for general use unless long term results have proven its value. PMID- 9411849 TI - [Physicians against land mines]. PMID- 9411848 TI - [3 years experience with endovascular stent prostheses in aortic aneurysm]. AB - Since August 1994 we changed our concept of the treatment of infrarenal aortic aneurysms at the Surgical Department of the Stadtische Kliniken Frankfurt Hochst. All patients morphologically suitable for endovascular aortic stenting were offered the new device. From the technical point of view stentgraft implantation is safe and its feasibility has been proven. The primary success rate was 88%. Perioperative mortality was 3.6%, primary leakage (10%) During follow-up thrombotic stentgraft occlusion (10%), renal artery closure (2%) and secondary leakage (10%) revealed a major problem. Most complications, however, could be treated interventionally without harm for the patient. We conclude form our results that good selection is the main key to avoid complications in the early phase of endovascular grafting. PMID- 9411850 TI - [Lipid reduction--benefit without risk?]. PMID- 9411851 TI - [Systemic sclerosis. Should care be centralized?]. PMID- 9411852 TI - [Is vaccination dangerous?]. PMID- 9411853 TI - [Osteoporosis]. PMID- 9411854 TI - [Osteoporosis. Bone mass, fractures, treatment and compliance]. AB - Following bone mineral mass measurement on 4,960 women, a follow-up study showed that the women who came from the rural area around Trondheim had higher, age adjusted bone mass and fewer fractures than those from the Oslo area, thus suggesting a geographic difference in bone mass and fracture risk. The differences in fracture rate were explained by the difference in mean bone mass in the two populations. The majority of the women with the lowest bone mass had started medical intervention, and the compliance was higher than 80%. It was highest among those women with the lowest bone mass. Because the patients had had the bone mineral measurements carried out on their own initiative, they are being followed up by their own doctors. However, there may be a need for closer individual follow-up of those who have the lowest bone mass. PMID- 9411855 TI - [Who is susceptible to osteoporosis? What is the significance of bone mass and clinical factors?]. AB - This paper reports on a questionnaire study based on 4,960 women who had had DXA measurements of bone mineral mass performed in Oslo and Trondheim. There was a great variation in bone mass in all age groups. The age-adjusted bone mass was a major contributing factor in all the fractures. Fractures in the femoral neck and other appendicular locations were most strongly related to femoral bone mass, while the spinal fractures were most strongly related to bone mass measured in the lateral projection of the spine. Clinical risk factors were only slightly related to bone mass and even less so to fractures. These relationships were of too little significance to adequately predict individual bone mass or the likely occurrence of fractures. PMID- 9411856 TI - [Treatment of planocellular carcinoma of the nasal vestibule]. AB - In the ICD-9 system, carcinomas of the nasal vestibule are classified together with carcinomas in the nasal cavity. As a rule, however, they are squamous cell carcinomas derived from the skin, and thus prognosis is better than in the case of squamous cell carcinomas derived from the mucosa of the nasal cavity. The article highlights essential features of brachytherapy. The authors present a critical assessment of the criteria for staging, and describe a specific patient material. Based on clinical experience and theoretical considerations, brachytherapy alone is recommended for T1-3 N0 tumours. In the case of T4 tumours external radiation therapy and brachytherapy combined is recommended for the primary tumour, with prophylactical irradiation towards regional lymph nodes. PMID- 9411857 TI - [Transient erythroblastopenia in children. Severe anemia with good prognosis]. AB - Transient erythroblastopenia of childhood (TEC) is an acute form of anaemia characterized by a transient red blood cell aplasia, of unknown cause, in the bone marrow. The incidence appears to be increasing. 16 patients were seen in our paediatric department during the period 1990 to 1996. The ages varied from two to 48 months. All patients had severe anaemia, the lowest mean haemoglobin values being 4.9 (2.2-7.8) g/100 ml. The reticulocyte count was low in 14 patients, whereas two patients had reticulocytosis. No underlying haematologic diseases were found. Ten patients were tested for parvovirus B19 infection, all of whom were serologically negative. Apart from transfusion of red blood cells in six patients, no therapy was given. Reticulocytosis, indicating beginning recovery, was observed after a mean interval of 11.8 days. This article gives a short overview of transient erythroblastopenia of childhood, a form of anaemia which can initially represent a diagnostic challenge. PMID- 9411858 TI - [Ulcerative colitis. Complications and sequelae in relation to the extent of the disease. A 10-year material]. AB - Patients with ulcerative colitis in our catchment area were followed over a ten years period with the focus on the relation between the extent of inflammation and complications. Total colitis was found in 180 out of 334 patients with known extension, while in 154 of the patients the inflammation was classified as left sided colitis. The frequency of severe colitis, colectomy and cancer was more or less the same as stated in earlier reports. We observed a difference, however, between the patients with extensive colitis and those with left-sided colitis as regards the course of the disease and the complications related to ulcerative colitis. The risk of liver disease, severe colitis and colectomy were significantly higher among the patients suffering from extensive colitis. Extraintestinal manifestations, on the other hand, were equally distributed among both groups of patients. The reasons for the observed differences are unknown. PMID- 9411859 TI - [Statins--the pattern of adverse effects with empahsis on mental reactions. Data from a national and an international database]. AB - We report on adverse drug reactions to statins recorded internationally and in Norway. The use of HMG-CoA reductase inhibitors (statins) has increased with a factor of 30 in Norway over the period 1989-96. Recently published clinical trials conclude that statins are safe; adverse drug reactions being infrequent and non-serious. The reactions observed are mostly increased hepatic enzymes and myopathy. Data from the Norwegian spontaneous reporting system, and from WHO's international database covering the period of 1988-95, includes reports of adverse drug reactions relating to other organ systems, such as the nervous system, the gastrointestinal tract, skin and cardiovascular organs. Psychiatric disorders represent 15% of the reactions to statins in the Norwegian database. Reactions include aggression, nervousness, depression, anxiety, sleeping disorders and impotence. The pharmacological mechanisms are not elucidated, but may be an effect of falling serum cholesterol. PMID- 9411860 TI - [Treatment of hypercholesterolemia with statins]. AB - The etiology of arteriosclerosis is a complicated interaction of many factors, among which aberrations in the cholesterol metabolism appear to play an important role. Several studies have shown that lipid-lowering therapy with statins improves survival and reduces complications in patients with coronary artery disease. Until recently, the role of cholesterol-lowering in patients with elevated cholesterol, but without diagnosed coronary disease, was not clear. New studies suggest that men in this group may benefit from treatment with statins. This article focuses on some of the problems connected with the widespread use of statins, namely: proper selection of patients, the safety of long-term statin therapy, economics, and the need for additional therapy and for the continuation of population-based general measures. PMID- 9411861 TI - [Systemic sclerosis. A diagnostic and follow-up program]. AB - Systemic sclerosis (scleroderma) is a rare chronic progressive multiorgan disease characterized by elevated collagen content in affected organs. The condition is classified in the group connective tissue diseases, and appears in both a systemic and a limited form. The rate of progression and the prognosis differ for the two forms. The heterogeneity of the disease makes systematic examination of the patients necessary in order to provide a correct diagnosis and classification as a basis for therapy and prognosis. Based on the low incidence and prevalence of the disease, the care of these patients should be a specialist task. At our department, a programme has been introduced consisting of clinical examination, immunological and biochemical analysis, histological examination and different imaging techniques, and with an interdisciplinary approach. This programme has made the management of these patients more standardized. PMID- 9411862 TI - [Magnetic tomography in diagnosis of breast implant rupture]. AB - At least 20,000 Norwegian women have silicone breast implants, either for breast augmentation or for reconstruction. One of the complications associated with breast implants is rupture of the implants. Magnetic resonance imaging (MRI) has been shown to be the most accurate imaging modality for evaluating the integrity of breast implants. Recognition of the different types of implants and the appearance of normal implants on MRI is very important for distinguishing these from intracapsular and extracapsular ruptures. Examples are shown of MRI findings in normal and ruptured implants. PMID- 9411863 TI - ["Bone markers"--when can they be useful?]. AB - There is an increasing interest in using deoxypyridinoline (D-Pyd) (Pyrilinks-D) and bone specific alkaline phosphatase (BAP) (Alkphase-B) or osteocalcin tests for detecting and monitoring risk persons for osteoporosis. Bone specificity is high, and the possibilities of detecting high bone turnover and fast bone losers in postmenopausal women is good, especially in women between 55 and 65 years of age. The tests are also useful for monitoring therapy, evaluating drug effectiveness and detecting poor compliance. The deoxypyridinoline (D-Pyd)-test must show a repeated high value prior to treatment (2 to 3 times over a period of one month), to allow for day to day variations. Because of the day to day variations in women's estrogen production (perimenopausal and early postmenopausal women), false low values may also be encountered. The use of "bone markers" to detect and treat fast bone losers could probably lead to considerable savings in the health budget and improvement in the quality of life for women worldwide. PMID- 9411864 TI - [Weight and weight gain in pregnancy]. AB - This article reviews different aspects of maternal weight before and during pregnancy and weight gain in pregnancy, e.g. causes of undernourishment (hyperemesis, anorexia nervosa and bulimia nervosa). Physiological weight gain during pregnancy is normally between 10 and 16 kg, representing 20% of the body weight before pregnancy. The increase in weight is usually lowest during the 1st trimester and greatest between the 17th and the 24th week of pregnancy. Low maternal weight at conception may cause low birthweight. Undernourishment may cause premature delivery or low birthweight, or both. There is an increased risk of gestational diabetes and macrosomia, as well as preterm delivery and hypertension in pregnant women who are overweight. There is also an increased risk of complications arising during general anaesthesia and operative delivery in severely overweight women. These women should be offered heparin or dextran as thrombosis prophylaxis where a caesarean section is to be performed. They should also be given antibiotic prophylaxis. A weight gain of between 7 and 12 kg reduces the risk of complications in overweight patients. PMID- 9411866 TI - [Psycho-cutaneous disorders in practice. Self-inflicted skin diseases of psychological origin]. AB - Patients with self-inflicted cutaneous lesions often consult practitioners or dermatologists rather than psychiatrists. The factitious skin lesions are pleomorphic, and may range from wounds, abrasions, blisters or burns, to relatively harmless patient-dependent aggravation of known dermatological diseases. The underlying psychopathology is highly divergent, ranging from socially induced stress behaviour to serious psychiatric disease with a high incidence of suicide. The basic psychological mechanisms of these diseases are poorly understood, and psychocutaneous disorders are difficult to treat effectively. The main challenge is to establish a trustful doctor-patient relationship, in which a joint therapeutic strategy can be established. The author describes six different psychocutaneous disorders in two selected cases, showing both the distinctions between the joint disorders and their common features. The need for constructive collaboration between practitioner or dermatologist and psychiatrist is emphasized. PMID- 9411865 TI - [Stress among air traffic controllers]. AB - Work stress was assessed by continuous logging of heart rate in 31 air traffic control personnel at seven airports in Norway. The results showed work stress within reasonable limits in all categories of air traffic controllers. Tests of psychomotoric functions in 36 operators revealed that all categories of operative personnel, but the air traffic controllers especially, emphasized accuracy at the expense of speed. Measurements of blood pressure in nine of the 33 air traffic controllers who had shown significantly elevated blood pressure in 1981 during a serious labour conflict revealed values below what was to be expected for their age group. PMID- 9411868 TI - [Fight against the abnormal. On medicine, power and adaptation stress]. PMID- 9411867 TI - [Use of curettage in the treatment of skin tumors]. AB - Many skin tumours can be diagnosed and treated with minimal equipment in general practice. The author describes the indications and technique of curettage for treatment of seborrhoeic warts, simple warts, and molluscum contagiosum, and basal cell carcinomas. PMID- 9411869 TI - [Land mines--the devil's seed]. PMID- 9411870 TI - [Aplastic crises in the course of hereditary spherocytosis]. PMID- 9411871 TI - [Should every patient with ulcerative colitis/Crohn disease be vaccinated against pneumococcal infection?]. PMID- 9411872 TI - [Iron supplementation in pregnancy]. PMID- 9411873 TI - [Continuous monitoring of stress during extreme strain]. PMID- 9411874 TI - [Latex allergy]. PMID- 9411875 TI - [Medicine and mass media]. PMID- 9411876 TI - [Challenge of the time]. PMID- 9411877 TI - [Our picture of the world]. PMID- 9411878 TI - [Remove acupuncture from the poster]. PMID- 9411879 TI - [Should not ambulance services in Norway be the responsibility of the community?]. PMID- 9411880 TI - [A proposal for new rules on continuing education of specialists in family practice]. PMID- 9411882 TI - [What happens to continuing education?]. PMID- 9411881 TI - [Resource utilization at intensive care units]. PMID- 9411883 TI - [Preventive measures should be gender-specific. Women and men belong to different health cultures]. PMID- 9411884 TI - [Smoking, body image and dieting habits among adolescents. A 3-year follow-up of adolescents aged 15-18 years]. AB - Several studies indicate that the differences in smoking habits which have hitherto prevailed between the sexes are decreasing, partly because of the increase in smoking among young Norwegian women. It has been suggested that concern about body image and weight gain is of particular relevance to women taking up smoking. In this article we present findings from a cohort study among 646 adolescents between the ages of 15 and 18 years. The results indicate that boys start smoking later than girls, and that concern about weight gain, body image and dieting plays a more important role in girls' than in boys' smoking habits at the age of 15. These factors are unrelated to boys' smoking habits at the age of 18, whereas they contribute significantly in predicting girls' smoking habits three years later. Furthermore, the results show that among girls, those who smoke are more concerned about gaining weight. Our findings emphasize the importance of implementing smoking prevention programmes at different age levels, using different motivating factors for boys and girls. PMID- 9411885 TI - [Victims of violence in the general youth population. A longitudinal study of risk factors]. AB - This paper presents new knowledge on risk factors involving violent victimization. The data stem from a representative survey of youths living in the Oslo area (n 447, response rate 86%). Questions on victimization experiences, drinking habits, alcohol problems, violence towards others and conflict-provoking behavior were first asked between the ages of 18 and 20 years (1993), and were repeated in the follow-up two years later. More men than women had been victimized. In both sexes there was a significant connection between alcohol problems and experiences as a victim. However, the consumption of alcohol and frequency of intoxication were only slightly related to such experiences. In men, both cross-sectional and longitudinal analyses showed that violence towards others had a much greater statistical effect than any of the variables relating to drinking behavior. In fact, the effect was greater than that of any other variable in the study. In women, previous victimization was, without doubt, the most significant predictor for future victimization. Stability over time of such experiences was also much higher for women than for men. As regards preventive efforts targeted towards victimization among youths, our findings suggest that a specific programme for each of the sexes needs to be developed and implemented. PMID- 9411886 TI - [Facial fractures. A life style disease among young men?]. AB - Interpersonal violence is often initiated by offensive verbal and visual expressions. Physical violence will therefore often be directed towards the face of the opponent. A retrospective analysis of 1,273 maxillofacial fractures at the University Hospital in Tromso during the period 1987-95 showed a steady occurrence of cases. A prospective study of 162 maxillofacial fractures at Ostfold Central Hospital in 1993 was compared with the material from Tromso. In both studies half of the patients were young men. Males aged 15-29 years were involved seven to eight times more frequently than the rest of the population. Violence among young men under the influence of alcohol was the predominant cause of the fractures. Young male assault victims often started the fight or actively encouraged it. "Participant" is often a more appropriate term than "victim". Some of those involved relapse into a lifestyle inflicting recurrent maxillofacial fractures. Domestic violence involving females is often disguised as accidents. Alternative methods of solving conflicts should replace the offending behaviour to prevent maxillofacial fractures. PMID- 9411888 TI - [Late effects of low-voltage electricity accidents. Rotator cuff tendinitis, hearing loss and neuropsychological dysfunction]. AB - This article describes the symptomatology of three patients following electrical accidents. The flow of the current was from hand to hand, voltage was 220/380 V, and duration was at least a few seconds for all patients. The development of symptoms was the same, and may be explained as a thermal effect of electricity on the tissue. Initially the patients experienced transient confusion, followed by stiff muscles after 1 to 3 days, and then pain in the muscle attachments and joints close to path of the current. This pain increased slowly during the first two weeks. Recovery was gradual, but often incomplete. The case notes showed that electrical accidents may be followed by chronic rotator cuff tendinitis. The clinical examination also revealed a hearing loss at about 2,000 Hz and above 4,000 Hz. The neuropsychological testing indicated a diffuse impaired function in only one patient. PMID- 9411889 TI - [Avulsion fractures of the anterior superior iliac spine. Lex coincidentia- again!]. AB - Avulsion fractures of the pelvic apophyses are fairly uncommon injuries. Most of these fractures occur in adolescent athletes. Traditionally they have been treated non-operatively with good results. We describe two cases of avulsion of the anterior superior iliac spine, which occurred coincidentally during a running competition at a Lille-hammer school in June 1996. The two fifteen years old boys were both treated with open reduction and internal fixation of the fragment. After three weeks they could walk normally. After 7-8 weeks they had resumed their usual sports activities, without any limitation of their sporting ability. Only a few case reports have been published dealing with operative treatment of avulsion fractures. It seems, however, that operative treatment might reduce the time to full recovery. For this reason, operative treatment of avulsion fractures should be considered for athletes and for other patients requiring a short period of rehabilitation. PMID- 9411887 TI - [Surgical injuries registered in the Norwegian system for patient injuries. Possibilities of the use of the material for quality improvement in Norwegian hospitals]. AB - 328 surgical "errors" reported to the Norwegian System of Compensation for Injuries to Patients were analysed in order to find out how the errors can be exploited for the purpose of quality improvement. In 8% of the cases the patients had been treated as emergency cases. 7% of the patients had been treated as out patients. 30% of the patients had become more than 15% permanently disabled as a consequence of the "error". The Norwegian System of Compensation for Injuries to Patients operates with five different categories of errors defined by medical specialty, of which surgery is one. We found that among "surgical errors" 16% of the patients had been treated by an anaesthetist or by a specialist in internal medicine, and 13% had been treated by a gynaecologist. There were several recurring "errors" such as nerve injuries and complications related to general atherosclerosis. A system for categorising errors with a view to quality improvement should be different from other systems of categorisation. We suggest a system based on not only five but all medical specialties. Data from such a system could be used to prepare "pedagogic reports" that can be sent to the managers of services and education in each medical specialty. Thus, by turning surgical errors into "medical treasures", the errors can be exploited to promote quality improvement. PMID- 9411890 TI - [Treatment of cholestatic liver diseases with ursodeoxycholic acid]. AB - A safe and effective medical therapy for patients with the chronic cholestatic liver diseases primary biliary cirrhosis (PBC) and primary sclerosing cholangitis (PSC) is lacking, and liver transplantation is at present the only curative treatment option. The effect of ursodeoxycholic acid (UDCA) in primary biliary cirrhosis has recently been investigated in several double-blind, placebo controlled trials. Treatment with ursodeoxycholic acid regularly improves the biochemical markers of liver disease. In some studies ursodeoxycholic acid delays the progression of primary biliary cirrhosis, whereas survival is not significantly affected. UDCA-treatment in primary biliary cirrhosis is now recommended in several countries, for example in the U.S. We suggest that UDCA treatment in Norway of patients with primary biliary cirrhosis should follow uniform guidelines. Ursodeoxycholic acid may also have a positive effect in primary sclerosing cholangitis. A Scandinavian multi-centre, placebo-controlled study of the effect of five-year long treatment of primary sclerosing cholangitis with ursodeoxycholic acid has recently started. The precise mechanism of the favourable effect of ursodeoxycholic acid in cholestasis is not understood. PMID- 9411891 TI - [Intensive care in Norway. A questionnaire study]. AB - A questionnaire was sent to five university hospitals, 16 central hospitals and eight large local hospitals. They were asked about the number of beds and the physicians employed in the intensive care unit (ICU), the use of some important intensive care procedures and treatments, registration and documentation procedures, and activity data for 1995. All the hospitals answered the questionnaire. The size of the unit varied from 5.5 to 7.0 beds and use of physicians varied from 1.3 to 5.2 (local vs university hospitals). Only six of the units use APACHE or SAPS severity scoring systems, and only three register work-load objectively. Definition of time spent in the ICU or on a ventilator varies. It is impossible at present to make a meaningful comparison of activity at the different intensive care units in Norway. If the situation is to be improved it is necessary to take the initiative for uniform use of definitions, objective scoring systems and registration of work-load. Such an initiative should come from the hospitals and the Norwegian Medical Association, in collaboration with the central health authorities. PMID- 9411892 TI - [Guidance in continuing education. Advantage or aggragation?]. AB - Educational counselling was introduced into specialist training in Norway in 1986. As educational counselling is regarded both as the basic structure of the training and the method, the Norwegian training programme has evoked interest in other European countries. Owing to scepticism and uncertainty as to the actual role of educational counselling in practical clinical work, the Norwegian Medical Association's Board for Specialist Education appointed a working group to look into this aspect of specialist training. In conclusion, educational counselling is widely used in all Norwegian training departments, but there is still a potential for improvement. PMID- 9411893 TI - [Poisonous mushrooms, mushroom poisons and mushroom poisoning. A review]. AB - Of 1,500 different types of Norwegian mushrooms, 60-100 are considered poisonous. Fatal intoxications occur very infrequently. Lack of knowledge of picking and preparing mushrooms and accidental or deliberate consumption are recognised causes of mushroom poisoning. Delayed onset of symptoms (> 5-6 hrs) indicates serious poisoning, and these patients must be admitted to hospital. Cytotoxic toxins (e.g. amatoxin, orellanin) cause serious damage to the visceral organs (liver, kidney) and require intensive treatment, including hemoperfusion. Neurotoxic toxins may cause dramatic, but less harmful peripheral or central symptoms affecting the peripheral and central nervous systems, including hallucinations. Some mushrooms cause gastroenteritis of low clinical significance within a few hours after consumption. Interaction between mushrooms and alcohol may lead to a disulfiram-like effect. Induced vomiting and activated charcoal are important initial therapeutic measures. The precise history of the patient and the collecting of mushroom remnants, including vomitus, may help to identify the particular mushroom. In Norway, the National Poison Information Centre may be contacted for further advice. PMID- 9411894 TI - [Pregnancy dermatoses]. AB - In the past, very similar and illogical names have been used for some of the dermatoses experienced during pregnancy, a situation which has confused diagnosis. The author suggests a simple classification where the dermatoses are organised into four groups: Pemfigoid Gestationis, Polymorphic Eruption of Pregnancy, Prurigo of Pregnancy and Pruritic Folliculitis of Pregnancy. The overview contains a brief description of the etiology, clinical signs, histology, prognosis and therapy for each type of dermatosis. PMID- 9411895 TI - [Mycosis fungoides. Early diagnosis and evaluation of treatment by polymerase chain reaction]. AB - We describe the case of a 60-year-old man who, after seven years with eczematous changes, developed erythrodermia, clinically suspect of mycosis fungoides. T-cell receptor gamma (TCR-gamma) gene rearrangements were retrospectively investigated by polymerase chain reaction (PCR). DNA from a formalin-fixed, paraffin-embedded punch biopsy showed a monoclonal pattern compatible with mycosis fungoides two years before the diagnosis was confirmed histopathologically. PCR was also used to evaluate the effect of different treatments. This case demonstrates the value of PCR for early diagnosis and evaluation of treatment of cutaneous T-cell lymphoma. PMID- 9411896 TI - [Human aspect of KOPF (Continuous Comprehensive Personal Binding) and patients' autonomy]. PMID- 9411898 TI - [The electronic medical record--is it here!]. PMID- 9411899 TI - [A urologist in cyberspace]. PMID- 9411897 TI - [Specialists' employment in a perspective of general practice]. PMID- 9411900 TI - [Old age, just a trouble? Medical periodicals call attention to the elderly and aging]. PMID- 9411901 TI - [Hemoglobin solutions. A step closer to artificial blood?]. PMID- 9411902 TI - [You are you and you are all right]. PMID- 9411903 TI - [Norwegian physicians--affluent and reputed, but not particularly happy]. AB - Norwegian doctors enjoy a high standard of living. A comparison of doctors' work environment and living conditions with those of the general population (Survey of Norwegian physicians' work environment and living conditions 1993 (Nord-Trondelag Health survey 1986, Survey of work environment 1989, General Household Survey 1991) shows that doing better does not necessarily mean feeling better. In almost all sex and age groups doctors score significantly lower than the rest of the population do on all our quality-of-life indicators. The mismatch may reflect the nature of their work. Although a larger share of doctors than others consider their work interesting and not physically tiresome, they work longer hours, and more them report feeling worn out, and that they have sleeping problems. Fewer of them describe their sociopsychological work environment as good. Possibly, as much effort should be put into improving doctors' jobs as into raising their salaries. PMID- 9411904 TI - [Job satisfaction among physicians employed in public health services]. AB - For several years the high turnover of doctors in municipal public health positions has been a cause of concern in Norway. More information on the causes of the higher turnover, and of factors that could promote greater satisfaction, could provide a basis for discussing what needs to be done. In order to find out the main sources of satisfaction, or dissatisfaction, and their relative importance, a questionnaire was mailed to all doctors in public health positions from 1988 through 1991. They were asked about 12 different aspects of their jobs and general family well-being. 218 (81%) of the doctors who had held such a position for the whole period ("the stable"), and 98 (87%) of those who had quit ("the quitters") responded. Sources of great satisfaction were opportunities for outdoor recreation and the well-being of the family. The doctors were least satisfied with the level of income and the heavy load of work. When asked to state the most important sources of satisfaction, both stable and quitters gave first priority to the professional content of the work and collaboration with colleagues. The stable also gave high priority to general family well-being. For the quitters, an important negative factor was the heavy work load. About 27% of the stable and 42% of the quitters thought the job required too much effort. More of the stable group than of the quitters felt professionally competent, and that they were doing a good job. On the other hand, more of the quitters than of the stable group felt they had little influence on the local health policy, and that the job was uninteresting. In the efforts to reduce the turnover of doctors in public health positions, the health authorities should consider the wider aspects of the job, not just the wage. To increase stability and job satisfaction, greater attention should be given to the professional content of the work, the need for professional collaboration, and the heavy work load. PMID- 9411905 TI - [Gastrointestinal amyloidosis. Differential diagnosis or a complication of inflammatory bowel disease?]. AB - A 77 year-old man developed intermittent diarrhoea and malabsorption. Endoscopic findings and preliminary histological examination indicated ulcerative colitis. Special staining of biopsies from the duodenum and colon revealed amyloid deposits. Classification of the amyloid fibril protein verified AL-amyloidosis, and the diagnosis primary idiopathic amyloidosis was made. Amyloid deposit in the gastrointestinal tract are a common feature of primary and secondary amyloidosis. The symptoms and findings are nonspecific and resemble those of chronic inflammatory bowel disease and ischemic colitis. Secondary amyloidosis can be seen as a rare complication of Crohn's disease and ulcerative colitis. Special staining is necessary to show amyloid deposit, and the distinction between primary and secondary amyloidosis requires immunohistochemistry. PMID- 9411906 TI - [Treatment chronic pain with amitriptyline. A double-blind dosage study with determination of serum levels]. AB - The aim of this study was to find an optimal analgesic dose of amitriptyline, and at the same time examine whether a therapeutic window existed for this analgesic effect. 85 patients with chronic, non-malignant pain were included in a double blind treatment regime with four doses of amitriptyline (10, 25, 50 or 100 mg). A blood sample was taken at steady state. The results showed 25 mg amitriptyline to have a good analgesic and sleep regulatory effect. The four different doses of amitriptyline did not show any significant difference in efficacy. No therapeutic window was found, but one cannot exclude that it exists. Low-dose amitriptyline, as a non-addictive drug, is a good alternative in the treatment of chronic pain, independent of co-morbid depression. PMID- 9411907 TI - [Lipoblastoma. A rare, benign tumor in children]. AB - Lipoblastoma is a rare, benign tumour of embryonal fat seen almost exclusively in infancy and early childhood. It occurs mostly in the extremities, but it is also seen in other parts of the body. The tumour may grow rapidly, and the fact that lipoblastomas show immature fat cells could lead to the wrong diagnosis of liposarcoma. Complete surgical excision appears to be the treatment of choice. A correct, preoperative diagnosis is possible in most cases. Two cases of lipoblastoma of the upper limb and one case in the scapular region are reported. PMID- 9411908 TI - [Clonazepam therapy of aberrant behavior in a mentally retarded man]. AB - In this study, the treatment of aberrant behaviour in a mentally handicapped male had failed. No link between the behavioural disorder and social circumstances was found. The subject had epilepsy. This was assumed to be a possible cause of the disorder and on this basis the antiepileptic medication was changed. Clonazepam was added to carbamazepin, and haloperidol was discontinued. The aberrant behaviour stopped. It is argued whether this was due to anticonvulsive effects; or anxiolytic, muscle relaxing and sedative effects. No definite conclusion is offered, although some findings could suggest the first interpretation. PMID- 9411909 TI - [A case of cowpox]. AB - A 37-year-old woman was admitted to the dermatology department at a regional hospital with a severe ulceration at the medial angle of the right eye. Virus culture yielded orthopoxvirus-like particles, later identified as cowpox virus. The clinical picture and virological diagnosis of cowpox are discussed. Emphasis is placed on the need for awareness among health personnel that such infections may well be encountered in an increasingly unvaccinated population. Guidelines for clinicians and for virology laboratories are given. Cats as a zoonoic source is discussed. PMID- 9411910 TI - [Primary hemochromatosis and dietary iron]. AB - Primary haemochromatosis is characterized by an unusually high degree of iron absorption resulting in the accumulation of excessive amounts of tissue iron. Excess stores of iron are removed by repeated phlebotomy. Health personnel and a number of patients with primary haemochromatosis have expressed their desire for advice on special diets to try and reduce the number of phlebotomies per year. This article gives advice on how patients with primary haemochromatosis can decrease their dietary iron intake and how they can put together meals to obtain low bioavailability, and therefore a lower iron absorption. The diet should be varied and be rich in bread and cereals, and fruit and vegetables. The amount of meat, Norwegian brown whey cheese (iron supplemented) and alcohol should be limited. Tea or coffee with meals will reduce iron absorption. Food rich in ascorbic acid (fruit and fruit juice) should be avoided with meals. Ascorbic acid supplements are not recommended. PMID- 9411911 TI - [Medical equipment--errors, causes and risks]. AB - Development in technology has introduced advanced technological apparatus into the health care sector. New technology has led to increased possibilities in diagnostics and treatment, but it also represents a substantial risk. A survey at the Lillehammer Hospital showed a relatively high rate of apparatus failure. However, only in a few cases was this of any consequence to the patient or the operator. The survey showed that preventive maintenance reduces the failure rate and that operator training and user checks reduce the risk related to medical apparatus. PMID- 9411912 TI - [Returned medicines as quality indicator for pharmacotherapy--what is left?]. AB - Unused medication accumulating in the homes of patients represents a medical, professional, financial and ecological challenge. Drugs returned to pharmacies show what kinds of medication are not being used. In our survey. 14 pharmacies registered all the medication returned during a three-month period. A total of 641 persons handed in 4,860 containers of medication during this period. A third of the containers were unopened and another third had more than half the contents left. The returned drugs were valued at approx. 450,000 NOK. Drugs used in cardiovascular and respiratory diseases constituted the largest part of the medication returned, both with respect to value and to volume. Drugs returned to pharmacies give an indication of the efficacy of pharmacotherapy. The results should prove suitable for reflection in multi-professional quality groups. PMID- 9411913 TI - ["To be used as directed by your physician"--reasons why patients do not use prescribed medicines]. AB - Research and practice indicate that a substantial amount of drugs prescribed for patients are never used. However, we do not know much as to why around 10% of the drugs paid for by the National Insurance are not used. The object of our survey was to identify and illustrate different reasons for this waste. On a questionnaire, 462 persons, who returned unused medication to 14 Norwegian pharmacies during a three-month period in 1994, indicated their reasons for not using the drugs. About half of these persons were also willing to participate in an in-depth interview. Using qualitative methodology, we grouped the reasons for having surplus medication into four main categories: Altered drug requirements- because of changes in the state of the patient's health or because of a new treatment regime. How the patients handled the prescribed drugs--influenced by the patient's knowledge, attitude and physical ability. Illness, social network and the complexity of the treatment were also contributing factors in this category. Coordination of the health services--related to how closely the various sections within the health care service worked together. Prescribing and distribution of drugs--related to rules and regulations, politics, technology, availability and the degree of economic consciousness among doctors and pharmacists. Both the professional and the socioeconomic aspects cited in this study call for reflection and may be used as a basis for further general discussion on the prescribing and handling of drugs, as well as the economic aspect of drug issue. They may also serve as an inspiration for local quality assurance measures and collaboration, especially between the various groups involved in the primary health care service. PMID- 9411914 TI - [How is it with the "elderly wave"]. AB - Data from Statistics Norway is used in this study of demographic development till 2020 in the population of the very old. There is a marked increase in the percentage of elderly women (80 years and over) in the population, from 2% in 1970 to nearly 5% in 1993. This is related to increased life expectancy and occurred parallel to a reduced rate in cardio-vascular mortality, which also occurred in men. However, in men this reduction is counter balanced by a serious increase, nearly 50%, in age-specific cancer mortality. The predictions for the population seem somewhat ambiguous, in particular as regards the percentage of elderly women, which is underestimated. There are significant regional differences with regard to the expected development in the population of people 80 years and older. On the whole, these differences do not seem to be related to mortality. PMID- 9411915 TI - [Changes among residents in homes for the aged in Bergen between 1985-96]. AB - Homes for the aged are intended for elderly people who, by and large, are able to perform basic activities of daily living (ADL) on their own, but who, in spite of this, need or wish to live in an institutional setting, where only minor nursing facilities are available. The present study focuses on the changes in mental capacity and in the altered need for nursing assistance with ADL functions which developed among residents at 13 homes for the aged in the city of Bergen, between 1985 and 1996. In 1985, the average age for residents was 84 years; and in 1996 it was 87 years. The average duration of stay was 30 and 38 months, respectively. There was a significant increase in the proportion of mentally impaired residents (28% and 35%). There was also an increase in the percentage in need of intensive nursing care (34% and 41%). The changes in residents living in the homes were not caused by dementia. It is concluded that homes for the aged are used increasingly for elderly in need of nursing care. There is an obvious mismatch between tasks and resources. PMID- 9411916 TI - [District geriatrics in Nordmore]. AB - Many aged Norwegians live in sparsely populated areas where access to geriatric assessment is limited. In 1990 a non-acute, ambulatory service was started in the Nordmore region. This article gives a description of the project. From 1990 to 1992 19 visits were made to six municipalities by a physician and a nurse from the out-patient clinic for the elderly at the local hospital. 59 patients were referred by general practitioners--mental impairment, general loss of function and assessment of possible rehabilitation being the most common causes for referral. Ten out of 11 GPs, all of whom had referred patients, and the leading district nurses in the municipalities expressed their satisfaction with the project through a postal questionnaire. PMID- 9411917 TI - [Geriatric assessment in Norway. Scandinavian guidelines for geriatric medicine]. AB - The Nordic countries have collaborated on reviewing and updating the guidelines of comprehensive case assessment in geriatric medicine. The Norwegian version of the document produced has recently been published as the official guidelines for the Norwegian Geriatrics Association. The document describes the status of geriatric medicine in the Nordic countries and summarizes the available documentation on its effectiveness and management. It also presents a detailed plan for carrying out geriatric case assessment. Scales for clinical use are evaluated and specific recommendations made with regard to their use in clinical practice. In this article the document is briefly described and commented on. PMID- 9411918 TI - [Euthanasia. An attempt to explain and define limits of the concept]. PMID- 9411919 TI - [Meeting a Greek psychiatric tragedy and contribution of the EU in relation to this]. PMID- 9411920 TI - [Anders Jahre's 1997 medical awards]. PMID- 9411921 TI - [Treatment of osteoporosis. Quality of life estimated without scientific or empiric basis]. PMID- 9411923 TI - [When were maternal-child health centers established?]. PMID- 9411922 TI - [Iron supplementation in pregnancy]. PMID- 9411924 TI - [Health personnel or hotel staff?]. PMID- 9411925 TI - [The potentials of computed tomography in the diagnosis of salivary gland diseases]. AB - Computer-aided tomography (CAT) was carried out in 68 patients with tumors of the salivary glands and 45 with sialolithiasis. CAT detected the tumors and located them, showed the type of the tumor (benign or malignant) and the tumor invasion. Sensitivity of CAT in tumors of the salivary glands was 97%, specificity 81.8%, accuracy 94.9%. In sialolithiasis CAT not only detected the concrements, but helped assess the status of the gland and the adjacent structures; its sensitively, specificity and accuracy were 95.5, 100 and 98.0%, respectively. PMID- 9411926 TI - [The clinical x-ray manifestations of a muscle imbalance in the temporomandibular joint and its treatment]. AB - The causes of dysfunction of the temporomandibular joint are analyzed, their clinical and x-ray symptoms enumerated, diagnosis and treatment described. The main cause of pain in this condition is the imbalance of masticatory muscles. PMID- 9411927 TI - [A new method for the surgical treatment of patients with paralysis of the tongue]. PMID- 9411928 TI - [The functional classification of velopharyngeal closure after uranoplasty]. AB - The new nasopharyngoscopic functional classification of palatopharyngeal joining is based on the results of examinations of 57 children with congenital uranoschisis after uranoplasty and 19 healthy children aged 5 to 14 years by nasopharyngoendoscopy. The classification defines the causes of palatopharyngeal insufficiency and for the first time takes into consideration the disagreement between the palatopharyngeal lock and articulation. The proposed classification helps select the most effective method of rehabilitation of the above patient population and helps follow up the time course of changes in the function of the palatopharyngeal lock under the effect of treatment. PMID- 9411929 TI - [Reconstruction of the visceral cranium by the replantation of a resected bone segment subjected to extracorporeal freezing in benign tumors and dysplastic processes]. AB - The new method for surgical treatment of benign tumors and dysplastic processes in the facial skull bones includes resection of the involved segment of the bone, its mechanical treatment, extracorporeal freezing, sterilization, and replantation. Biophysical studies revealed that 10-min freezing of the preparation in liquid nitrogen followed by slow defrosting kills tumor cells. Animal experiments demonstrated excellent osteoplastic properties of thus treated replants. Based on the results of treatment of 29 patients, the authors defined the indications for such treatment and demonstrated its high efficacy in benign tumors and dysplastic processes in the facial skull. PMID- 9411930 TI - [Reconstruction of the external nose in patients with posttraumatic defects and deformities of the bones of the nasoethmoidal-orbital area]. AB - Examinations and treatment of 52 patients (28 men, 17 women, and 7 children) with posttraumatic deformations of the nasoethmoidal-orbital complex permitted the authors to distinguish the main types of deformations and develop methods for surgical treatment. Autotransplants of the skull vault were used for correcting the deformations in all cases. The proposed methods of treating posttraumatic defects and deformations of the osseous and cartilaginous parts of the nose help attain stable functional and cosmetic results and are less time-consuming. PMID- 9411931 TI - [The psychological assessment of patients with congenital and acquired maxillofacial deformities in the pre- and postoperative periods]. AB - Complex rehabilitation of 20 patients with congenital and acquired maxillofacial deformations included psychological examinations using the finish-the-sentence test, Taylor's, Eysenck's [correction of Isenk's], Leonhard-Smisek, and Dembo Rubinstein tests. Depressive states were detected in 4, neurotic reactions with depressive incorporations in 16 examinees. After surgery the manifestations of depression decreased in 60% of patients. Psychological studies helped improve the efficacy of surgery due to a favorable impact on the personality. PMID- 9411933 TI - [Enhancing the biocompatibility of dentures made from polymethylmethacrylate by using hydroxyapatite]. AB - Twenty or 50 weight-percent of hydroxyapatite (Polystome) was added in suspensions of AKR-15 copolymer and acid monomers, then the mixtures were molded and subjected to thermal treatment. Admixtures (supercritical CO2) were extracted from the composites in parallel with infrared spectroscopic measurement of the concentration of methylmetacrylate toxic monomers. Hydroxyapatite in 20% concentration decreased the release of toxic substances from the composite, whereas 50% concentration of the agent did not produce this effect. Three patients with intolerance of acrylic plastic were effectively treated using this method. PMID- 9411932 TI - [A trial of the use of rulid, sumamed and makropen in the combined treatment of generalized periodontitis at a stage of exacerbation]. AB - In vitro study of the antibacterial activity of macrolide antibiotics azitromycin (sumamed), midicamycin (macropen), roxitromycin (rulide), and erythromycin demonstrated their high activity towards clinical strains of bacteroids, fusobacteria, peptostreptococci, streptococci, and corynebacteria. These antibiotics were effective in the treatment of 62 adult patients with severe and moderate generalized periodontitis. Rulide and sumamed were the most effective, macropen and erythromycin were inferior to them. PMID- 9411934 TI - [The enhancement of the efficacy of patient orthodontic treatment based on the mathematical modelling of prospective implant designs]. AB - Opportunities to increase prosthetic treatment efficiency by means of applying osseointegrated implants of traditional and prospective types are analysed. A technique of mathematical modelling of the interaction between implants and jaw bone is exposed. The numerical model and applied program, developed on the basis of finite element method, enabling to analyse a stress strain state of the bone and to determine extreme safe loads on implants are described. It is shown, that the most loaded zone is a layer of the compact bone directly contiguous to the neck of the implant. That is in good agreement with the results of clinical research, according to which just this zone has the highest percentage of complications. Methods for further optimisation of implants and prosthetic structures for the purpose to perfect the techniques of prosthetic treatment are discussed. PMID- 9411935 TI - [The use of ceramic onlays for restoring the occlusal surface of the teeth]. AB - Computer-aided milling of ceramic insertions helped more effectively repair the masticatory surface of teeth, minimize the incidence of errors, rule out the laboratory stage of making the insertions and the technological errors made at this stage, retain physical and mechanical characteristics of ceramics, optimize marginal adherence of the insertion to the tooth, retain the height of the lower part of the face, and thus prevent the maxillodental dysfunction. PMID- 9411936 TI - [The monitoring of the level of body fluoride intake in children participating in 3 milk fluoridation projects in Russia]. AB - In Russia three milk fluoridation projects have been implemented since 1994 (cities Smolensk, Voronezh and Maykop) covering more than 10,000 children living in low fluoride areas and having high risk of caries development both in temporary and permanent dentitions. Both preliminary and monitoring studies of fluoride excretion with urine and fluoride consumption were done. Total number of subjects--663 children 3-6-years of age attending kindergartens in these three cities; 2602 urine samples were collected, twice analyzed for fluoride content and processed in accordance with the method approved by the Russian Ministry of Health. 24-month monitoring of milk fluoridation projects in Russia did really show and confirm three main facts: the concentration of fluoride 2.5 mg/l in F milk products for children 2-6 year-olds in low fluoride areas of Russia is really optimal concentration when daily milk consumption is about 200 ml; there is no such phenomena as detectable accumulation of metabolic fluoride under physiological conditions even after 2 years of regular fluoridated milk consumption; developed technique of fluoride consumption determination proved to be a very useful tool for planning and monitoring preventive programmes based on systemic use of fluoride. PMID- 9411937 TI - [A comparative assessment of the efficacy of different types of sealants for the prevention of caries in the permanent teeth of children in the oral hygiene system]. AB - Three hermetics are assessed: chemically hardened Delton, light-hardened Estiseal, and composite Evikrol. The study was carried out in 126 children aged 6 years. The decrease of the increment of dental caries depends on the retention of hermetics on the occlusion surface of the teeth, and the efficacy of caries prevention in permanent teeth is much higher if hermetic sealing of fissures and fossae is combined with local fluorine prophylaxis and oral hygiene. All types of hermetics can be used to prevent permanent teeth caries, but chemically hardened ones should be preferred. PMID- 9411938 TI - [Clinical and diagnostic problems in cancer of the facial skin]. PMID- 9411939 TI - [The incidence dynamics of dental caries and periodontal diseases in the population of different territorial and administrative areas of Tajikistan]. PMID- 9411940 TI - [The use of the preparation Solpadeine in dental practice]. PMID- 9411941 TI - [Software support for automatic (computerized) work stations for dentists]. AB - The authors describe the software for a basic computer unified system of working places for dentists specializing in oral, dental surgery and orthodontics with diaries and special entries for each section. The computer system head physician- registration office--primary examination room--rooms of dental treatment, physiotherapy, and x-ray examination--is intended for use as part of local computer network of dental clinics in the MS DOS and Windows systems. Application of the system improves the labor efficiency by up to 35% and the financial efficacy by almost 30%. It also warrants legal protection of both physicians and patients. PMID- 9411942 TI - [The problem of hepatitis infection in dentistry]. AB - The incidence of hepatitis B is increasing all over the world. The parenteral route of infection is one of the principal. A considerable number of cases are caused by infection of patients at therapeutic institutions, including dental clinics. Medical workers are at risk of the infection, too. One main cause is neglect of sanitary antiepidemic measures: manipulations without gloves, veils, protective eyeglasses; lack of disposable instruments, poor cleaning, disinfection, and sterilization of hardware. Awareness and alertness of medical personnel is an important factor. Since dental care is a most prevalent kind of medical service, urgent measures are needed to decrease the risk of nosocomial infection with hepatitis for both patients and staff. PMID- 9411943 TI - [The ecological aspects of stomatological pathology]. PMID- 9411944 TI - [An analysis of the mortality of patients with suppurative-inflammatory diseases of the maxillofacial area]. AB - The frequency of acute purulent inflammatory diseases of maxillofacial area and their complications (thrombosis of the cavernous sinus, abscess of the brain, sepsis) were investigated. These complications are responsible for 0.56% of lethal outcomes. Early diagnosis and intensive treatment of such conditions are a must. PMID- 9411945 TI - Human schistosomiasis in the Senegal River Basin. PMID- 9411947 TI - North American experience with rabbit anti-human thymocyte globulin (thymoglobulin) and rabbit anti-human thymocyte serum in solid organ transplantation (1989-1996). Proceedings of a symposium. Mont Tremblant, Quebec, Canada, January 31, 1997. PMID- 9411946 TI - The ParaSight-F test for detecting treatment failure. PMID- 9411948 TI - [The polymorphism of the kappa-casein gene and its connection to economically valuable traits in cattle]. AB - The literature data on milk protein (particularly kappa-casein) polymorphism study in different cattle breeds are represented. Allele variants of kappa-kasein are associated with lactation and have an influence on the composition of milk and its properties, including cheese-making properties. The importance of ways of kappa-casein genetic variants use in concrete selection programs are discussed. PMID- 9411949 TI - [The characteristics of the chromosomal level of variability in mice]. AB - The analysis of the order of chromosome involving to interchromosome associations by the type of Robertsonian translocation (RB) in 28 cell populations in relation with three factors: their origin from different mouse lines (CBA/Ca, C3H/He, C57BL/6, BALB/c), direction of cytodifferentiation (fibroblasts and leucocytes) and the stage of its disruption (minimal, maximal and medial tumorigenicity) was carried out. The influence of all three factors on the frequencies of individual chromosome involving in RB was revealed. PMID- 9411950 TI - [The estimation of the degree of genetic divergence in Bovidae by DNA restrictase analysis]. AB - Restrictase DNA analysis was performed in seven species of Bovidae family- European (Belovezh) bison, Belovezh-Caucasian bison, European-American bison, mountain Caucasian bison, American bison, domestic bull, and domestic sheep. DNA was digested with MspI and TaqI restriction endonucleases. The products obtained were end-labeled and electrophoretically separated in poliacrilamid gel. DNA fragments consisting of highly repetitive genomic sequences were detected as a set of bands corresponding to fragments between 464 and 67 bp in length. All forms of Bison genus analysed were characterized by the identical sets of bands. The band patterns of bulls is similar to these of the bison, whereas the patterns of sheep is rather different. These data indicate that American bison and European bison geographically isolated forms of the same polytypic species. PMID- 9411951 TI - [A comparative analysis of Ungulata species by different molecular genetic markers (proteins, RAPD-PCR)]. AB - The investigation of genetic interrelation between a number of Artiodactyla and Perissodactyla species with the use of different types of molecular-genetic markers (proteins, RAPD-PCR) were carried out. The marker-specific features of interspecific relations and their similarities on the groups of markers of both types were revealed. The distinctions between interspecies genetic relations and ones estimated from the phylogeny on the determined group of different types of markers were observed. It was supposed that these discrepancies may be related with common selection factors and involving this marker group in selection in some species. PMID- 9411952 TI - [The muzzle and biochemical genetic markers as supplementary breed characteristics in cattle]. AB - The comparative analysis of characteristics of three different cattle breeds (Brown Carpathian, Pinzgauer, Red Polish) on the 5 molecular-genetic markers and 5 muzzle dermatoglyphic types was carried out. It was indicated, that one characteristic can not be use as a breed-specific one but only their complex. The main aspect of search of this complex is the use of characteristics which mark different structure-functional systems of whole organism. PMID- 9411953 TI - [Old age--a trump or a joker?]. PMID- 9411954 TI - [The geriatric ward in Denmark]. PMID- 9411955 TI - [Functional assessment of the elderly. Practice and clinical research]. PMID- 9411956 TI - [Wound healing and wound treatment in the elderly]. PMID- 9411957 TI - [Methodological challenges in measurements of functional ability in gerontological research]. AB - This article addresses the advantages and disadvantages of different methods in measuring functional ability with its main focus on frame of reference, operationalization, practical procedure, validity, discriminatory power, and responsiveness. When measuring functional ability it is recommended: 1) Always to consider the theoretical frame of reference as part of the validation process. 2) Always to assess the content validity of items before they are combined into an index and before performing tests for construct validity. 3) Not to combine mobility, PADL and IADL in the same index/scale. 4) Not to use IADL as a health related functional ability measure or, if used, to ask whether problems with IADL or non-performance of IADL are caused by health-related factors. 5) Always to analyse functional ability separately for men and women. 6) To exclude the dead in analyses of change in functional ability if the focus is on predictors of deterioration in functional ability. PMID- 9411958 TI - [Preventive home visits to the elderly. An evaluation of controlled interventional studies]. AB - Since 1st July 1996 communities in Denmark are legally obliged to offer preventive home visits to elderly people. Twelve randomized controlled trials examining the effects of this kind of intervention have been published. The trials differ widely with respect to their design, the intervention and the evaluation. The review of the twelve trials shows that preventive home visits have a beneficial effect on elderly people and that hospital and nursing home admissions decrease. The underlying cause for this is not clear. However, the studies indicate that a) health care personnel have to undertake these visits, b) social and medical intervention is necessary, and c) visits have to be made on an ongoing basis. The further implementation of preventive home visits to elderly people in Denmark should be closely monitored. PMID- 9411959 TI - [Hearing in a geriatric perspective]. AB - To assess whether hearing rehabilitation of older people can be improved by co operation between the audiology and geriatric departments and the home service, 139 old and frail audiological patients were allocated to three groups with three different fitting procedures: 1) conventional fitting including verification of acoustical gain in the patient's ear; 2) home-fitting by hearing therapists, and 3) home-fitting by a specially trained geriatric nursing assistant, the home help also being present. Outcome was assessed by the ordinary questionnaire mailed to hearing aids users three to four months after fitting and by a geriatric evaluation procedure. The response rate in the conventionally fitted group was highly unsatisfactory (36%) and too small for further data-analysis. In the educational group a tendency was found towards better manipulation skills and significantly higher hearing aid use. However, the response rate was lower than in the geriatric group (71% compared to 81%), and no knowledge of hearing aid use was registered in the home service by this procedure. In the geriatric group a correlation was found between practical ability and use and satisfaction with the hearing aid. However, two thirds of the group were dependent on lasting help for the handling of the aid. Most patients in this group were already known by the hospital and home service, and the individual home help showed an interest in learning about hearing aid use. Home fitting by a joint audiological and geriatric effort in collaboration with the home help has proven feasible and valuable to both patient and home help. Extended co-operation is recommended between the health care and the social sector concerning hearing aid use. PMID- 9411960 TI - [Physical fitness of Danish men and women aged 50 to 80 years]. AB - Maximal power in sustained work in originally randomly selected men and women, born in 1914, was studied five times between the ages of 50 and 80 years in a longitudinal design. Of the originally 514 men and 461 women in 1964 living in the Western suburbs of Copenhagen, 23 men and 18 women performed a bicycle test at age 50, 60, 70, 75 and 80. The mean annual decline in body mass adjusted maximal power in sustained work (W/kg) was 1.43% in the 18 men and 1.64% in the 23 women. Based on "cross-sectional" comparisons of all subjects tested at any age, the mean annual decline in men was 1.56%; in women the corresponding figure was 1.80%. When the results of the "longitudinal" and "cross-sectional" analyses were compared with each other, a rather similar picture of the age-related decline in maximal power was obtained, especially in women. In the longitudinal data only moderate (women) or zero (men) correlations were observed between the submaximal test results at the ages of 50 and 60 years and the maximal test results at higher ages. The physical work load at the age of 50 years had no significant correlation with maximal power at that age or thereafter. There were only minor changes in mean body height, body mass and BMI during the follow-up. PMID- 9411963 TI - [Gene therapy against aging: possible or not?]. PMID- 9411962 TI - [Predictive value of the screening instrument "Mini-assessment of nutritional status"]. AB - The purpose of the study was to evaluate the predictive capacity of the Mini Nutritional Assessment method (MNA), by means of data from the Danish part of the SENECA survey (1988), and the follow-up study from 1993. Using the MNA, 171 persons between 70-75 years of age were classified according to their nutritional risk as either being "well-nourished", comprising 78.4% or "at risk of malnutrition" comprising 21.6%. A total of 115 persons participated in the follow up study. The participation rate in the follow-up study was significantly lower in the risk group (p < 0.01). The 13 Danes judged "at risk of malnutrition" had had a significantly higher occurrence of acute diseases (p < 0.05), need of help (p < 0.05) and weight loss (p < 0.001) in the preceding years. However, no significant differences were found in hospitalization rates and physician visits between the two groups. In conclusion, the results indicate that the MNA is capable of identifying a group of 70-75 year-old Danes who are at risk of certain nutrition-related health problems. PMID- 9411961 TI - "RAI"--an international system for assessment of nursing home residents. AB - The Resident Assessment Instrument (RAI) is described. The RAI consists of three basic components: 1) the Minimum Data Set (MDS) is a collection of items covering all the areas one would expect when assessing the needs of elderly people and developing a care plan, 2) the Resident Assessment Protocols are a series of protocols which guide the assessor through the best practice of care planning for the common problems faced by the elderly and 3) the Resource Utilization Groups is a classification system based on MDS to predict resource use at the level of the individual resident. The goal of RAI is to improve the quality of care. It also enables national and international comparisons between long term care residents. RAI was used at 65 nursing homes in Copenhagen in 1992-93 and 3451 residents were examined. The reproducibility and validity of the instrument was tested and found to be good. Results from the use of RAI in Denmark is presented. We conclude that RAI is useful in assessing residents at nursing homes, because the collected data triggers actions that improve the quality of care. PMID- 9411964 TI - [Acut confusion in elderly patients]. PMID- 9411965 TI - [Accelerated course of operations--why and how?]. PMID- 9411966 TI - [Nutrition of hospitalized patients]. PMID- 9411967 TI - [Lung function tests]. PMID- 9411968 TI - [Malignant intestinal obstruction]. PMID- 9411969 TI - [Current development in chromosome analysis]. PMID- 9411970 TI - [Mortality and cause of death among Danish physicians 1973-1992]. AB - This study examines mortality rates of Danish doctors and describes pattern and causes of death for the period 1973-1992. The study comprises 21,943 medical doctors, 6012 of whom were women. At the end of 1992 there were 2387 recorded deaths. The doctors had lower mortality rates than the general population. A significant lower mortality was seen for male medical specialists compared to general practitioners. A gender-difference was seen among the youngest doctors with the female doctors suffering a considerably higher mortality than the male doctors did. Both sexes showed SMR below unity for cancer, circulatory diseases and other natural causes. Mortality due to lung cancer was particularly low. The suicide mortality was increased for both sexes, in particular because of an increased number of suicides by poisoning. Compared with the general population the doctors' mortality was low, but the mortality from external causes was increased, mainly due to an excess number of suicides. PMID- 9411971 TI - [Increased urea synthesis in patients with active inflammatory bowel disease]. AB - Patients with active inflammatory bowel disease are often reported to be in negative nitrogen balance. Therefore, we examined basal and amino acid stimulated urea synthesis in 11 patients with active inflammatory bowel disease and in 10 patients with non-active disease. A primed continuous infusion of an amino acid mixture was given from t = 1 h to t = 5 h; during the first and the last two hours no amino acid infusion was given. Urea nitrogen synthesis rate was quantified independently of changes in blood amino acid concentration by means of the functional hepatic nitrogen clearance, i.e. the linear slope of the regression of urea nitrogen synthesis rate on blood amino acid concentration. Basal and amino acid stimulated urea nitrogen synthesis rate as well as functional hepatic nitrogen clearance were elevated twofold in the patients with active disease. No differences between the two groups were observed as regards basal or stimulated plasma glucagon, cortisol, catecholamines and serum levels of interleukin-1 alpha, interleukin-1 beta, tumor necrosis factor-alpha and interleukin-6. The results show that liver function related to conversion of amino-nitrogen to urea is increased and may contribute to the less efficient nitrogen economy in patients with active inflammatory bowel disease. PMID- 9411972 TI - [Abuse of benzodiazepines among heroin addicts in Copenhagen]. AB - Abuse of benzodiazepines among heroin addicts in Copenhagen. The aim of this study was to investigate the extent of benzodiazepine abuse among heroin addicts, and to examine whether the heroin addicts with and without benzodiazepine abuse differed socially and psychiatrically. Social and medical information was drawn from the records for 98 heroin addicts who were registered for treatment at the institution Distriktscenter Vestre between 1.2-30.9.1994. Information about psychiatric admissions was taken from the Danish Psychiatric Register. The results showed that the heroin addicts with and without benzodiazepine abuse do not differ with regard to psychiatric data and basic demographic data. However, addicts with concomitant benzodiazepine ab-use were worse off concerning housing conditions, employment criminal records and amount and type of heroin abuse, and are thus in need of a more concerted treatment effort. PMID- 9411973 TI - [The diagnosis of "smoker's lung" encourages smoking cessation]. AB - In a controlled randomised trial we analysed whether the use of the term "smoker's lung" (Danish: "rygerlunger") instead of chronic bronchitis when talking to patients with chronic obstructive lung disease (COLD) changed their smoking habits. Fifty-six smoking patients with COLD were allocated to either intervention (n = 25) or control groups (n = 31). In the intervention group the lung disease was designated smoker's lung in all communication with patients about their illness and in the control group traditional terminology was used. All patients were given the same medical treatment and the same encouragement to stop smoking. One week after discharge 57% had stopped smoking in the smoker's lung group vs 26% in the control group (p = 0.028), at three months 50% vs 19% (p = 0.027) and at one year 40% vs 20% (p = 0.148). Referring directly to the cause of a self-inflicted illness may be an effective way of discouraging risk behaviour, at negligible cost. PMID- 9411974 TI - [Burn wounds after resuscitation of a newborn girl]. AB - In resuscitation of an asphyctic newborn baby, a chemical hot pack was used for maintaining sufficient body temperature. The child lay for 45 min on the covered hot pack, which had a peak temperature of 54 degrees C. This led to a third degree burn (5% of BSA). The child needed split skin grafting. Control at four months of age showed a well-taken skingraft and the child was developing normally. We conclude that chemical hot packs should not be used for newborn babies unless the peak temperature in the packs is less than 44 degrees C. PMID- 9411975 TI - [Mealworm allergy]. AB - A 24 year-old female employed in a pet shop developed occupationally related asthma, rhinoconjunctivitis and contact urticaria caused by exposure to the yellow mealworm (Tenebrio molitor Linnaeus) sold as food for birds and reptiles. A wholebody extract of the mealworm showed positive prick test and histamine release. PMID- 9411976 TI - [International variations of the survival of cancer patients. Comparison between USA and Canada]. PMID- 9411977 TI - [Dissection, autopsy, postmortem examination]. PMID- 9411978 TI - [Antiviral combination therapy against HIV]. PMID- 9411980 TI - [Heart rate variability in evaluation of the autonomic nervous system. A review]. AB - Analysis of heart rate variability is increasingly used for testing the function of the autonomic nervous system in cardiovascular disease and for the diagnosis of autonomic neuropathy. In cardiovascular disease, long-term data collection (several hours) is primarily used to describe the static activity in the different branches of the autonomic nervous system. In the diagnosis of autonomic neuropathy, short-term forced variations in heart rate are employed in order to describe the dynamic capacity of the parasympathetic nervous system. In the subsequent data-analysis several different principles may be used, and the purpose of this review is to describe these principles and their use in clinical and experimental settings. PMID- 9411981 TI - [Meta-iodobenzylguanidine scintigraphy of the heart]. AB - Iodine-123 labeled metaiodobenzylguanidine (MIBG) is a noradrenaline analogue. The tracer is thought to follow the same mechanisms of uptake into cardiac sympathetic nerve endings as noradrenaline. MIBG-scintigraphy can thus be used to noninvasively image the cardiac sympathetic nervous innervation. The uptake and distribution of MIBG in the heart are changed in various cardiac diseases, and MIBG-scintigraphy has become a new research tool especially for the evaluation of patients with myocardial infarction and ischaemia, congestive heart failure and ventricular arrhythmias. This review article focuses on the cardiac diseases where MIBG-scintigraphy has contributed to a better understanding of the pathophysiology and improved clinical evaluation of the patients. PMID- 9411979 TI - [Functional MR imaging of CNS]. PMID- 9411982 TI - [Age at menarche and first sexual intercourse. A cross-sectional study of 17-year old Danish women]. AB - The age at menarche and first intercourse was analyzed from data obtained from a questionnaire sent to 1500 17 year-old randomly selected Danish women (76% responded). The average age at menarche was 13.37 years (SD: 1.24). The median age at the time of the first intercourse was 16.6 years. Late sexual debut was observed for girls who were relatively old at menarche and for those with a higher level of education. Apparently the age at menarche and at first sexual intercourse has not changed significantly in recent years. PMID- 9411983 TI - [Examination and treatment of musculoskeletal diseases in general practice investigated by medical audit]. AB - The general practitioner has a central role in the investigation and treatment musculoskeletal diseases, because he often is the first to see the patient. Correct initial care can save days lost through sickness and prevent chronic courses. In May 1995, 173 general practitioners registered every patient contact with complaints from the musculoskeletal system. The registration continued for three weeks following an audit model used in Denmark among general practitioners (in total 6869 contacts). After the registration four follow-up meetings took place. The two main results were 1) frequent conventional X-rays of osteoarthritis, and 2) prescription of non-steroidal antiinflammatory drugs to every fourth contact with musculoskeletal complaints. The follow-up meetings revealed some non-professional motives for sending the patient to X-ray examination and for prescribing non-steroidal anti-inflammatory drugs. The results will form the basis for clinical guidelines for handling some of the musculoskeletal diseases. PMID- 9411984 TI - [Fatal hyperthrombotic condition complicating nephrotic syndrome]. AB - A twenty-year old woman was admitted to hospital with impaired consciousness and a left hemiparesis. A cerebral CAT-scanning was reported normal. Convulsions and respiratory insufficiency made intubation and mechanical ventilation necessary. Laboratory examination showed an extremely low concentration of albumin in plasma (109 microM) and gross albuminuria was present. No other signs of renal failure occurred. The patient deteriorated during three days and died with signs of cerebral incarceration, despite efforts to reduce intracranial pressure. Autopsy showed a large thrombus of the sagittal sinus as the presumed cause of death. No thrombus was found in the renal vein. The association between nephrotic syndrome and thrombosis is discussed. PMID- 9411985 TI - [Cervical extraosseous Ewing sarcoma]. AB - A case of extraosseous Ewing's sarcoma is presented. The clinical, morphological and imaging characteristics of this rare neoplasm are briefly discussed. PMID- 9411986 TI - [Autonomic dysfunction in chronic liver disease]. PMID- 9411987 TI - [Prevention of hepatitis B]. PMID- 9411988 TI - [Prevention of hepatitis B, one more comment]. PMID- 9411990 TI - [Renography in the investigation of hypertension]. PMID- 9411989 TI - [Basic ethics]. PMID- 9411991 TI - [Continuous care in pregnancy and childbirth]. PMID- 9411992 TI - [A clinical physiologist and nuclear medicine on the Internet]. PMID- 9411994 TI - [What do people talk about in Danish hospital elevators?]. AB - Hospital elevators make up a part of the public space of hospitals which is used by both employees, patients, and patients' relatives. We performed an observational study to ascertain the frequency of ethically problematic conversations between employees in the elevators of three Danish hospitals. We defined a conversation as problematic if it breached confidentiality or if it could raise doubts about the professional or ethical standard of the hospital or its employees. Based on this definition we found that problematic conversations occurred in 18 out of 201 elevator-trips where more than one member of the hospital staff was present in the elevator (9%, 95% confidence limits 5-14%). The most common types of problems were breaches of confidentiality (6 cases), complaints about stressful work (4 cases), and complaints about the organisation (3 cases). PMID- 9411993 TI - [Is smoking during pregnancy a cause of testicular cancer?]. AB - Smoking during pregnancy is suggested as a cause of the increasing incidence rates of testis cancer in Denmark, particularly for the age group between 25 and 50. Testis cancer is associated with lower birthweights, as is smoking during pregnancy. Danish women have rates of everyday smoking amongst the highest in Europe, and the highest rate of smoking during pregnancy in Scandinavia, followed by Norway, Sweden and Finland. Incidence rates for testis cancer show the same sequence. The increasing incidence of testis cancer in Copenhagen was temporarily halted among men bom between 1939-1945 during the wartime tobacco shortage. The maternal generation aged about 25 years during the war years shows a corresponding delay in the increasing incidence of lung and bladder cancer, with a lag-time of respectively 20 and 30 years. The author recommends long-term followup of newborn boys prenatally exposed to tobacco. PMID- 9411995 TI - [Occurrence of renal artery stenosis in patients with peripheral arteriosclerosis and hypertension]. AB - The aim of this study was to evaluate the prevalence of renal artery stenosis in patients with clinical signs of peripheral vascular disease and hypertension. One hundred patients, mean age 69 years (range 45-88) with symptoms and clinical signs of severe peripheral ischaemia, underwent aortography to determine the degree of peripheral vascular disease and possible renal artery stenosis. History of claudication and measurement of systolic distal blood pressure and calculation of the Ankle Brachial Index was used to define the severity of peripheral vascular disease. Thirty-one percent had renal artery stenosis (14% greater than 50% reduction in luminal diameter). In a subgroup of patients with hypertension and peripheral vascular disease (n = 74), 34% had renal artery stenosis. In the subgroup of patients with renal artery stenosis, 81% had hypertension. Among patients with renal artery stenosis and lumen reduction of more than 50%, 93% had hypertension (p < 0.001). In conclusion this study shows that the combination of peripheral vascular disease and hypertension is an important clinical clue for renovascular disease. Examination for renovascular disease in this population should be considered, since the prevalence of the condition is high. Furthermore, examination for renal vascular disease in this population is mandatory, before treatment with angiotensin converting enzyme inhibitors is initiated, since treatment might lead to serious renal function impairment. PMID- 9411996 TI - [p16 and p53 genes transferred with the help of adenovirus to induce apoptic tumor cell death]. AB - Based on studies of the cell cycle in tissue cultures of cancer cell lines and cells from normal tissue interference in the regulatory network of pRB/cdk4/cyclin D1/p16 in combination with the tumour suppressor gene p53 was investigated. It was shown that overexpression of p53 and p16, but not p53 on its own, induced apoptotic cell death only in tumour cells. Gene transfer of the same two genes to tumours transplanted subcutaneously in mice also caused fast regression of some of the tumours. PMID- 9411997 TI - [Attendance pattern at an open casualty ward in Greater Copenhagen]. AB - To analyse the use of a casualty department in Copenhagen, we prospectively analysed all contacts to the casualty department at Hvidovre Hospital during a period of four weeks. There were 3908 contacts and 3851 of these were accessible for registration with a full record (98.5%). The mean intake was 130 patients per 24 hours. Sixty eight percent were seen within 24 hours of the accident or onset of serious symptoms of illness, 22% were admitted by ambulance. A total of 680 patients (18%) came to the hospital rather than their general practitioner for reasons of convenience. Only one third of the patients presented an orthopaedic problem. Twenty-five percent of the patients presented a problem, which properly should have been dealt with by their general practitioner. Eight hundred and forty-seven (22%) of the contacts were unnecessary. Like most other hospitals in Denmark, all patients at Hvidovre casualty department are evaluated by a junior orthopaedic surgeon. By establishing a possibility for primary evaluation and proper distribution of patients by an older and more broadly educated colleague, it should be possible to eliminate approximately 30% of the attendances from the emergency department and there by give better time and treatment for those in real need of help. PMID- 9411999 TI - [Obesity, dieting and eating disorders]. PMID- 9411998 TI - [Reduced sense of taste as a complication of the laryngeal mask use]. AB - We report a unilateral lingual nerve paralysis following the use of a laryngeal mask airway in a 73-year-old man with hypertrophy of the prostate undergoing a TUR-P. The operation was performed in general anaesthesia using propofol, fentanyl, alfentanil and ventilation with 100% oxygen. The operation was uncomplicated and had a duration of two hours and twenty minutes. A week later the patient perceived a reduced sense of taste. Six months later the situation was unaltered. PMID- 9412000 TI - [Is obesity contagious?]. PMID- 9412001 TI - [Gestational diabetes mellitus]. PMID- 9412002 TI - [Malaria diagnosis]. PMID- 9412003 TI - [Rifabutin. An antibiotic against mycobacteria]. PMID- 9412004 TI - [The laparoscopic treatment of abdominal cryptorchidism in adults]. AB - Three patients aged 18, 25 and 31 years with abdominal cryptorchidism and concomitant diseases were treated laparoscopically. Macroscopic amd microscopic evidence said in favor of removal of the testis. In one case this removal was done in line with removal of the dysplastic kidney. In another case plastic surgery was conducted of the inguinal hernia on the side of cryptorchidism. PMID- 9412005 TI - [A clinical trial of the treatment of patients with benign prostatic hyperplasia using the alpha 1-adrenoblocker alfuzosin]. AB - Alfuzosin (dalphaz) has been tried in 32 patients with benign prostatic hyperplasia (BPH). The treatment lasted for 14 weeks: 2 weeks of placebo and 12 weeks of alfuzosin. The drug was given in a daily dose of 10 mg (5 mg twice a day). The response was registered in 87.5% of cases. Improvement of urination occurred by all the parameters of IPSS scale (the overall IPSS dropped by 10.5 scores). Dalphaz had a good hypotensive effect: high blood pressure lowered to normal, normal pressure did not change. Dalphaz was well tolerated in all the cases except one when an allergic eruption broke out. PMID- 9412006 TI - [The use of new technologies in the endoscopic electrosurgery of benign prostatic hyperplasia of large dimensions]. AB - The technique of electrovaporization, a novel low-traumatic method, has been practiced in the Research Institute of Urology (Moscow) for the treatment of benign prostatic hyperplasia (BPH) since 1995. This technique is based on the standard transurethral resection (TUR) being superior in the absence of intraoperative bleeding. Its application, however, is restricted to hyperplasia of small and moderate size. But TUR, in its turn, provides more rapid removal of the tissue. These advantages of the two techniques were combined in the treatment of extensive BPH. Compared to TUR, standard TUR combination with electrovaporization proved much more effective. Progressive techniques in endoscopic surgery comprise also use of novel resection electrodes which differ from the previous analogues by thickness, shape and coating. When used in electroresection, such electrodes provide simultaneous removal of the tissue, electrovaporization and coagulation of the underlying tissues. The above technical innovations allowed advance in the quality of endoscopic interventions, widening indications for endoscopic electrosurgical management of large-size BPH and decreasing the incidence of relevant complications. PMID- 9412008 TI - [The role of free-radical lipid oxidation in the pathogenesis of chronic prostatitis]. AB - The study of antioxidant activity (AOA), levels of molecular products of free radical oxidation and ceruloplasmin in secretion of the prostate and blood plasma in 30 healthy males and 90 patients with chronic prostatitis has revealed multidirectional shifts in AOA, prostatic secretion and ceruloplasmin plasma levels in patients with chronic prostatitis. Parallel shifts in AOA and ceruloplasmin correspond to the view on ceruloplasmin as leading antioxidant in blood plasma. High ceruloplasmin concentrations in chronic prostatitis may be considered as compensatory-adaptive response aimed at compensation of local ceruloplasmin deficiency and AOA in the inflamed prostatic tissue. Artificial rise of ceruloplasmin content in the blood flow reduced clinical and laboratory signs of chronic prostatitis. PMID- 9412007 TI - [Sexual readaptation after the surgical treatment of benign prostatic hyperplasia]. AB - Sexual function was studied in 818 patients with benign prostatic hyperplasia (BPH) before and after surgical treatment of this disease. Before surgery, sexual activity was absent in 276 examinees. After surgery 4.3% of them retained erection, 95.7% remained impotent. 542 patients before operations were sexually active. Surgical treatment of BPH (transurethral resection, transvesical adenomectomy) creates grounds for deterioration of sexual function and risk of erection loss. Thus, 77 operated patients had no erection, 176 had weak libido, 159--insufficient erection, 244 retrograde ejaculation, 188 painful orgasm. Transurethral resection led to a complete loss of copulative function in 5.3% of patients, transvesical adenomectomy--in 9.9%. Sexual readaptation after transurethral resection and transvesical adenomectomy has been improved due to a special complex developed by the authors. This complex consists of 14 therapeutic and prophylactic procedures. PMID- 9412009 TI - [The establishment of the I-PSS in the CIS countries taking into account its cultural and linguistic features. International Prostate Symptom Score]. AB - I-PSS adaptation to cultural and linguistic features of CIS population has been made in two stages. The 1st stage consisted of adaptation of the questionnaire's text and its check up by the specialists. At the 2nd stage a random sample of 46 males at the age 60 to 80 years were tested and retested. Reliability of the results has been proved in repeated tests and comparison of the adjusted questionnaire with its English-American variant. The authors propose to use adapted questionnaire I-PSS which meets international requirements in the countries members of CIS. PMID- 9412010 TI - [The urological complications of contraception using intrauterine coils]. AB - Being a foreign body, intrauterine coil causes decubitus and inflammation of the adjacent tissues. Long-term carriage of the coil may give rise to endometritis, myometritis, parametritis, salpingo-oophoritis, tubo-ovarian inflammatory infiltrates. These infiltrates invade retroperitoneal pelvic fat and may obstruct pelvic ureters. Ureteral obstruction may bring about ureterohydronephrosis, pyelonephritis and renal calculi. The coil may be also responsible for chronic pyelonephritis. The authors have treated 64 females aged 18-45 years with urological complications due to intrauterine coils which stayed from 6 months to 14 years. 34 of them presented with attack of acute pyelonephritis, 29 with renal colic and acute pyelonephritis, 26 with renal calculi. To arrest renal colic and attack of acute pyelonephritis ureteral catheterization and renal pelvis drain were performed in 31 patients. One patient has undergone ureterolithotomy. 8 patients rejected removal of the coil and had recurrent renal colics and acute pyelonephritis attacks. Removal of the coil arrested pyelonephritis and lithogenesis in the kidney. In one case of coil removal there was injury to the uterine cervix and urinary bladder eventuating in vesicovaginal fistula. PMID- 9412012 TI - [Ultrasonic monitoring of transurethral prostatic drainage in chronic prostatitis using the Intraton-4 electrostimulator-aspirator]. AB - A detailed ultrasonic picture of the prostate was provided by the rectal device of Pie Medical-200 unit for 1066 patients with chronic prostatitis before and in the course of drain therapy using electrostimulator-aspirator Intraton-4. The leading ultrasonic diagnostic symptoms of chronic prostatitis are represented by various microspaces and hypoechogenic zones in different prostatic parts which reflect activity of chronic inflammatory process. The severity of the disease was determined primarily by the presence in the prostate of "true microabscesses" (microcavity of the irregular form) and pseudomicroabscesses (drop-shaped microspaces) which can be successfully drained irrespective of their number, size, location and characteristics of the content. The evidence gave grounds for design of the scheme describing pathogenesis of chronic prostatitis and its reverse dynamics in the course of drain therapy. These reverse changes closely resemble stages of ureterohydronephrosis. PMID- 9412011 TI - [Urotractin in the treatment of an infection of the kidneys, urinary tract and prostate]. AB - Clinical and bacteriological effects of urotractin are analysed for 60 patients with infectious-inflammatory diseases of the kidney, urinary tracts and prostate treated in 3 urological clinics of Moscow. The drug was given 10 days in a dose 400 mg twice a day. Urotractin demonstrated high efficacy against both gram negative and gram-positive bacteria. The results were the same in in- and outpatients. PMID- 9412013 TI - [The enzymatic activity of the seminal plasma in ejaculates of different fertilities]. AB - Enzymatic ejaculates activity was studied in 15 fertile and 65 subfertile samples of human sperm. The activity of alkaline phosphatase (EC 3.1.3.1), alpha-amylase (EC 3.2.1.1), gamma-glutamyl transferase (EC 2.3.2.2), creatine kinase (EC 2.7.3.2), total lactate dehydrogenase (EC 1.1.1.27) and lactate dehydrogenase-1 was compared to spermogram findings according to 14 morphological and physicochemical criteria of fertility. The enzyme activity depends on functioning of the testes, seminal vesiculae, prostate, bulbourethral glands and is involved in formation of fertility. The findings allowed elucidation of the role of synthesis and some function of spermoplasma enzymes. Practical applications of the above data are discussed in the treatment of male infertility and to raise fertilizing power of the sperm in artificial insemination. PMID- 9412014 TI - [The immunochemical determination of the concentration of organ-specific ejaculate proteins in the differential diagnosis of chronic inflammatory diseases of the male reproductive system]. AB - Concentrations of lactoferrin (LF), alpha-microglobulin of fertility (AMGF) and specific thermostable alpha-glycoprotein (STAG) which are markers of the prostate, seminal vesicles and testiculi in ejaculate, respectively, were determined by Ouchterlony's immunodiffusion method in 148 patients with chronic prostatitis (group 1), 26 patients with combination of prostatitis with chronic epididymitis (group 2) and 22 healthy controls (group 3). After that the patients were given prostatilen. It was found that high pretreatment values of LF registered in groups 1 and 2 lowered close to those in the controls. This shows elevated LF to be a diagnostic sign of prostatitis. In group 1 AMGF was lower than in controls, in groups 1, 2 and 3 STAG levels were similar. PMID- 9412015 TI - [Neurosomatic parallels in children with neurogenic bladder dysfunction]. AB - Clinical, urodynamic and neurological examinations of 30 children with hyperreflex dysfunction of the bladder gave evidence for clinical symptoms of natal trauma sequelae in 23 of them. Diagnosis of polyvisceral functional changes and chronic urinary diseases was made significantly more frequently in children with neurological symptoms. High occurrence of neurological symptoms of natal subclinical cervical traumas in children with hyperreflex dysfunction of the bladder suggests their pathogenetic relations. Hemodynamic changes of the cerebral vessels with affection of the venous outflow may cause hypoxia of the hypothalamo-hypophyseal structures with emergence of symptom complex of vegetovascular dystonia and marked neurogenic dysfunction of the bladder. The authors conclude on validity of pathogenetic neurological therapy to coordinate the activity of spinal and supraspinal centers regulating function of the bladder. PMID- 9412016 TI - [Invasive transitional-cell cancer of the bladder in a young man]. PMID- 9412017 TI - [Allergic vasculitis in benign prostatic hyperplasia combined with a latent prostatic carcinoma]. AB - Histological examination of the removed prostate from a 66-year-old patient discovered glandular hyperplasia, atypical hyperplasia and small focus of poorly differentiated (latent) adenocarcinoma. There were nodular lymphocytic infiltrates in the adenoma, in the peripheral parts of the node there was destructive thrombovasculitis and productive granulomatous vasculitis in the presence of diffuse lymphohistiocytic infiltration. Vascular changes resembled those in Winiwarter [correction of Winivater]-Berger disease. Vasculitis is thought an allergic reaction. PMID- 9412019 TI - [The use of terazosin hydrochloride (Hytrin) in the treatment of patients with benign prostatic hyperplasia]. PMID- 9412018 TI - [The membrane phospholipid peroxidation and Ca-dependent ATPase activity of the microsomal fractions isolated from rat renal tissue in thermal ischemia with and without alpha-tocopherol protection]. AB - The author studied the effects of 30-min heat ischemia of rat kidneys on the level of malonic dialdehyde (MDA) and Ca-dependent ATPase activity of microsomal fraction isolated from the cortical substance in the presence and absence of antibiotic alameticine and ortovandate in the incubation medium and protective action on Ca-ATPase activity of rat pretreatment with alpha-tocopherol (TP). It has been demonstrated that thermal ischemia induces inhibition of Ca-ATPase activity of microsomes resistant to vanadate. Administration of TP reduced MDA level, enhanced Ca-ATPase microsomal activity in the presence of alameticine against inhibition of enzymic activity in the absence of alameticine. This indicates a rise in the true enzyme activity under decreasing membrane permeability in conditions of diminishing activity of lipid peroxidation in response to TP effects. PMID- 9412020 TI - [Postload hematuria in virtually healthy miners]. AB - 21 healthy miners with hematuria occurring after working hours (the study group) and healthy miners without hematuria (controls) were examined before and after work. Compared to controls before work, the study group had low activity of plasma renin (APR) and intensive glomerular filtration, low urine levels of Na+ and its rapid clearance. Compared to controls after work, hematuria subjects had low blood pressure, elevated APR, activated vasopressor prostanoids, low level of blood K+. PMID- 9412021 TI - [The diagnosis and surgical treatment of cavernous hemangiomas of the liver]. AB - The authors have examined 80 patients with cavernous hemangiomas. The disease was diagnosed on the basis of an ultrasonic investigation, computed tomography and laparoscopy. Solitary hemangiomas were diagnosed in 78 patients, multiple ones in 2. In 64 patients the size of hemangiomas was more than 5 cm, in 16 patients they were less than 5 cm. Radical resections of the liver were made in 32 patients (59.3%), palliative--in 22 patients (40.7%). Complications after the liver resections took place in 15.5% of the patients, lethality was 6.2%. Complications following the palliative operations were noted in 72.7% of the patients, lethality was 9%. PMID- 9412022 TI - [Russian surgeon N. D. Monastyrskii (on the 150th anniversary of his birth)]. PMID- 9412023 TI - [Sympathetic tonus and lumbar sympathectomy in arteriosclerosis obliterans of the arteries of the lower extremities]. AB - The Doppler laser flowmetry was used in order to study the microcirculation in the skin, muscles and bone marrow at different stages of obliterating atherosclerosis of the lower extremity arteries. Progress of the disease is characterized by a decrease of the absolute indices of blood flow in combination with disturbed regulation of vascular tension. The lumbar sympathectomy causes intensification of microcirculation not only in the skin, but also in the muscular and bone marrow tissues. PMID- 9412024 TI - [A methodology for the objective assessment of trauma severity (III. The assessment of the severity of the state of wounded and injured patients)]. AB - The article describes original scales for the estimation of the severity of state of patients with wounds and traumas. The scale I has been developed for using at the first level of the objective estimation of severity of the wounded's state during the medical sorting. It allows the sorting to be performed very exactly and has advantages over the analogues known in the literature. At the second level the scale II is used which is meant for the estimation of the severity of the wounded's state on admission to the hospital, while at the third level the scale III is used in the dynamics of treatment. These scales are necessary due to different qualitative and quantitative signs used during making the diagnosis and different values of the diagnostic signs for establishing the probability of lethal outcomes and complications on admission to the hospital and a day after performing a complex of resuscitation measures and intensive therapy. A distinctive feature of these scales is their polycriterial character, i.e. each code of the severity is calculated according to two criteria: probability of the development of lethal outcome and complications of the injury. The scales are based on simple symptoms and can be used under the military field conditions and in extreme situations. PMID- 9412025 TI - [A new symptom of acute appendicitis in school-age children]. AB - In addition to the well-known classical symptoms of acute appendicitis the author of the article has revealed a characteristic feature of the disease in 63.2% of children from 6 to 14 years of age. It is the flexion and extension of the right foot toes during the palpation of the right iliac area. The degree of the symptoms is proportional to the severity of inflammation of the vermiform process. PMID- 9412026 TI - [The differential diagnosis of posttraumatic ossifications in the area of the elbow joint in children]. AB - Clinico-roentgenological observations of 64 patients aged from 4 to 14 years with posttraumatic ossifications in the elbow joint area, of whom 25 were operated upon, enabled the authors to compare the data obtained with the well-known signs of osteogenesis of another etiology. The case history and x-ray examinations are thought by the authors to be the most important methods allowing the nature of this osteogenesis to be differentiated. The main distinctive signs of the posttraumatic ossifications are the development of their roentgenological stages in the special chronological succession. PMID- 9412027 TI - [A new method for evaluating the rheological properties of the erythrocytes in surgical patients with endogenous intoxication]. AB - An examination of 30 patients against the background of using both the operative and conservative methods of treatment was performed in order to study the rheological properties of blood and selection of the most informative tests for diagnosis of microcirculatory disturbances in diseases of the gastrointestinal tract and gastroduodenal area. The authors propose methods of investigation of deformability and viscosity of erythrocytes in patients with the surgical diseases in question. The methods are simple, quite handy for any surgical hospital and polyclinic laboratories. In addition to being used in investigations of the aggregative ability of erythrocytes and viscosity of the whole blood, these methods can be of use in express diagnosis of rheological disorders. The endogenous intoxication in patients with the diseases in question was found to be accompanied by deep rheological disturbances. The deformability of erythrocytes and their viscosity index which can pass ahead of the shifts in other parameters characterizing the hemocoagulative and rheological properties of blood are changed. A conclusion is drawn that the parameters of the erythrocyte deformability and viscosity can be an additional criteria of the adequacy of therapy used for patients with the diseases in question. PMID- 9412028 TI - [Anemia in burns and the possibilities for its correction]. AB - A cytological investigation of erythropoiesis and the peripheral blood was performed in 66 patients with a severe burn disease. It was established that the ultraviolet photomodification of the autoblood in complex with detoxication optimized hemopoiesis and allowed the nontransfusion correction of burn anemia to be performed at the expense of using the reserves of medullary hemopoiesis. The maintenance of basic parameters of the peripheral blood at the adaptational level requires 20% less erythrocyte-containing preparations of blood which naturally reduces the risk of post-transfusional complications. PMID- 9412029 TI - [The surgical treatment of chronic duodenal obstruction in combination with chronic pancreatitis and tumors of the pancreaticoduodenal area]. AB - Results of 102 surgical interventions were analyzed. In 39 patients chronic duodenal obstruction was associated with benign diseases, 63 patients had malignant tumours. The chronic duodenal obstruction resulted from diseases in the pancreatoduodenal area in 80 patients, in 22 patients it proved to be the cause of chronic pancreatitis. The adequate surgical correction of the duodenal obstruction is believed to be sufficient for normalizing the pancreas functions in patients with primary chronic duodenal obstruction. Direct surgical interventions are preferable for secondary chronic duodenal obstruction. Prophylactic formation of gastroenterostomy during the biliodigestive surgery can relieve the symptoms of the developing duodenal obstruction and allows to avoid another operation at the advanced stage of chronic duodenal obstruction. PMID- 9412030 TI - [Pancreaticoduodenal resections "at the height" of profuse hemorrhage from a tumor]. AB - The authors share their experiences with observations of profuse bleedings from the tumour of the pancreas head into the duodenum lumen. The clinical picture of this complication and surgical methods of treatment are described. Both patients recovered. PMID- 9412032 TI - [The surgical treatment of retroperitoneal phlegmons]. AB - Based on the experience with treatment of 112 patients with retroperitoneal phlegmons of different etiology the authors distinguish and describe the diffuse and complicated forms of the disease. In treatment of phlegmons special importance is attached to the optimal surgical access. The article describes main principles of performing sanitation of the suppurative foci of the retroperitoneal space, variants of the aspiration-irrigation methods of treatment of the suppurative process. The authors believe that their original differentiated surgical method using different variants of the aspiration irrigation drainage is effective in treatment of patients with retroperitoneal phlegmons. This method of treatment gave 23.2% lethality while after the traditional methods of treatment 90% of the patients died. PMID- 9412031 TI - [Intestinal perforation in tuberculosis]. AB - For 5 years the authors observed isolated abdominal tuberculosis in 6 patients. Five of them who were urgently operated upon had perforations of the tuberculous ulcers of the intestine. A specific character of the inflammation was confirmed by histological investigations. Four patients are described in detail. One of them had six perforated tuberculous ulcers of the jejunum. All these patients died. Recommendations are given how to improve the diagnosis of abdominal tuberculosis and treat patients with this severe disease. PMID- 9412033 TI - [The use of a perfluorocarbon emulsion in the local treatment of wounds complicated by a surgical infection]. AB - The article shows effectiveness of the local application of Perftoran in complex treatment of purulent wounds of soft tissues in 50 patients. Perftoran has no necrolytic and antibacterial properties and fails to make the terms of the 1st phase of the wound process shorter. Perftoran maintains the intensified development of young cell elements at different periods of healing of the purulent wounds, decreases the propagation of microorganisms in them, lessens the wound square surface, results in earlier development of marginal epithelialization and formation of the elastic cicatrix. PMID- 9412034 TI - [Sepsis. The polemical aspects of the problem]. PMID- 9412035 TI - [Generalized forms of the inflammatory reaction and of surgical infection. Current problems in the terminology and delimitation of concepts]. PMID- 9412036 TI - [Non-Hodgkin lymphomas of the gastrointestinal tract. The surgical aspects of the problem]. PMID- 9412037 TI - [Semen Semenovich Girgolav (1881-1957)]. PMID- 9412038 TI - [International Congress of Northern Countries and Regions on Problems of Old Age and Actual Questions of Geriatric Surgery. Kondopoga, Karelia, June 26-28, 1997. Abstracts]. PMID- 9412039 TI - [The effect of a paravertebral novocaine block on acid formation in the stomach]. AB - The dependence of acid-formation in the stomach on the blockade of sympathetic innervation of the acid-formation area was investigated in 38 patients in whom the paravertebral novocaine blockade was made at the ThVII-ThIX level in order to cup off the pain syndrome. The control group included 7 patients without diseases of the organs of the hepato-pancreatico-duodenal zone. This blockade was found to sharply increase the basal acid-formation in healthy people. In patients with the high acid-formation it leads to greater elevation of acidity. It was shown that the disturbed functional balance between two parts of the vegetative nervous system responsible for the acid-formation function of the stomach was one of the causes of higher acid formation in patients with peptic ulcer of the gastroenterostomy and with chronic ulcer of the duodenum. It seems to be reasonable to investigate disturbances of the function of sympathetic innervation of the acid-formation area of the stomach for determining the optimum volume of surgical interventions in such patients. PMID- 9412040 TI - Student numbers and university funding. PMID- 9412041 TI - Settlement of insurance claims. PMID- 9412042 TI - [99m-Tc-Technetryle mammascintigraphy: clinical protocol and first experience in clinical diagnosis of breast cancer]. AB - Analyzing the results of 99mTc-Technetryle mammascintigraphy in 41 females with breast cancer indicated that polypositional planar mammascintigraphy ensures a high sensitivity (over 88%) in revealing a primary nodule in T1, (more than 95%) in large tumors at above 95% specificity. No nodal accumulation of 99mTc Technetryle was observed in controls (with fibrocystic mastopathy and suspected coronary heart disease. Mammascintigraphy showed a 85% sensitivity in recognizing axillary lymph nodal metastases too and ascertained that 14 patients had suptaclavicular and subclavian lymph nodal metastases. Therefore, 99mTc technetryle mammascintigraphy is the method of choice in early detecting a tumorous process, primarily lymphogenic metastasis in breast cancer, and in evaluating its extent. PMID- 9412043 TI - [X-ray computerized tomography in the differential diagnosis of gastric ulceration]. AB - The results of more than 3000 gastric examinations were used to study the potentialities of X-ray computerized tomography in the differential diagnosis of benign and malignant gastric ulcerations. CT evidence for benign and malignant gastric ulcerations is outlined. The place and role of CT in the diagnostic algorithm for gastric ulcerations are defined. In the authors' opinion, CT is a supplementary study and should be used purposefully after preliminary X-ray, endoscopic, and morphological examinations in difficult clinical situations associated with the differential diagnosis of gastric ulcerations. PMID- 9412044 TI - [Computed tomography and ultrasonography in the diagnosis and treatment of pancreatic diseases]. AB - Seventy three percutaneous punctures and catheter drainages were performed in 68 patients in 1992-1996. Visual control under CT was made in 32 patient and that under ultrasonography in 41. Diagnostic aspiration fine-needle biopsies of pancreatic lesions in 33 patients shower 5 adenocarcinomas, 2 cysts with signs of malignancy, 2 pseudocysts, 1 pancreatic adenoma, 23 chronic pancreatitis. Percutaneous catheter drainage in 15 patients with pyogenic complications of acute pancreatitis yielded good results. Successful external drainages of pyogenic pancreatic pseudocysts followed by sclerotherapy were made in 21 patients. There were recurrent cysts in 2 patients. This method is effective and safe for the diagnosis and treatment of pancreatic diseases and should be widely put into clinical practice. PMID- 9412046 TI - [Potentialities of digital low-dose x-ray apparatus in the diagnosis of heart and lung diseases]. AB - The paper presents experience in clinically testing a digital low-dose X-ray apparatus. X-ray studies were performed in 169 patients: 140 with cardiovascular abnormalities and 19 with lung diseases. The findings show the high quality of images of thoracic aorta and lung structures. The radiation dose received during an examination is many times lower than at routine X-ray study and fluorography, which makes it possible to use the apparatus in mass surveys of contingents at risk for tuberculosis. PMID- 9412045 TI - [Potentialities of echography in the diagnosis of uncomplicated acute appendicitis in children]. AB - Ultrasonography was performed in 97 children admitted to hospital for suspected acute appendicitis. According to the results of primary surgical examinations and the magnitude of clinical manifestations, all the patients were divided into two groups. Group 1 comprised 53 patients with clinically diagnosed acute appendicitis and with a worded indication for an emergency surgical intervention; this group was formed for describing the ultrasonic semiotics of different types of acute appendicitis in children. Group 2 (n = 44) was enrolled to study the differential diagnostic potentialities of echography in the primary diagnosis of acute appendicitis in children with acute stomach ache and they are diagnostically unclear. All echocardiographic findings are supported by intraoperative data and pathohistological evidence. The ultrasonic semiotics of different forms of uncomplicated acute appendicitis is described, the optimal procedure has been developed for examining children with this pathology. The results of the study confirm the high informative value of the proposed algorithm of an echographic examination and sensitivity of the method as to the patients of this group. Thus, the sensitivity of the echographic method in uncomplicated acute appendicitis was 88%. PMID- 9412047 TI - [To rare manifestations of lymphogranulomatosis]. PMID- 9412048 TI - [Examination of gastrointestinal tract using the Russian x-ray contrast media BAR VIPS based on Barium sulfate]. PMID- 9412049 TI - [X-ray characteristics of heart failure in patients with acute myocardial infarction]. AB - The results of routine chest X-ray made in 250 males aged 22-69 years who had acute myocardial infarction were evaluated. The data were compared with the results of ECG, echocardiography, Judkins coronarography, and left ventriculography. The X-ray signs of pulmonary venous hypertension in acute myocardial infarction, even not followed by cardiomegalia suggest lower left ventricular myocardial contractility. In this connection, the significance of follow-up X-ray monitoring becomes higher. In 25% of the young patients (aged 22 40 years) with prior acute myocardial infarction, the dimensions of the heart may be in the normal ranges even in the presence of X-ray signs of venous congestion. If there are no signs of mitral regurgitation in patients with ischemic heart disease, the enlarged left atrium may be regarded as an indirect X-ray sign of reduced left ventricular contractility. The extent of necrosis in patients with myocardial infarction exerts an impact on hemodynamic changes in the lung. PMID- 9412050 TI - [The University of X-ray Laboratory Assistants. Lesson 2. Milliampere-second]. PMID- 9412051 TI - [X-ray fluorographic screening diagnosis of central and peripheral lung cancer]. PMID- 9412052 TI - [Diagnosis of endocardial electrode microdislocation at endocardial pacing]. AB - Signs of microdislocation of an endocardial electrode (EE) were observed in 56 (14.4%) of 390 patients with an implantable pacemaker in the postoperative period. The microdislocation was diagnosed from electrocardiographic evidence for no response of a pacing pulse and from fluctuations in the pacing threshold, and from X-ray findings of the thickened and ill-defined outlines of a distal EE with preserving cardiac stimulation. The microdislocation preserves high parameters, as evidenced by a PM-20 M digital analyzer. PMID- 9412053 TI - [The delivery of psychiatric care to servicemen during wartime]. PMID- 9412054 TI - [Problems in the moral and psychological support of the training of military physicians at a military college]. PMID- 9412056 TI - [Experience in the medical support for operations to eliminate the aftermath of an accidental spill of rocket fuel components]. PMID- 9412057 TI - [The status of and ways to improve the reproductive health of servicewomen]. PMID- 9412058 TI - [Qualified and emergency specialized surgical care for those with wounds to the extremities]. AB - Experience of organization of the surgical care in the military hospital to 438 wounded in extremities during armed conflict in Republic of Chechnya is generalized. Maximum reduction of stages of medical evacuation of the wounded in extremities, approaching of the qualified and urgent specialized surgical care directly to the region of battle actions, use of opportunities for it one-moment rendering corresponded to principles of the modern military-medical doctrine. Due to realization of the requirements of the doctrine life of many wounded ++ was saved, terms of treatment, medical and social rehabilitation are reduced. Besides lethality, treatment cost and numbers of transferring to the reserve from the Armed Forces were reduced. PMID- 9412059 TI - [The acupuncture of biologically active points in the treatment of primary corneal stromal dystrophies]. PMID- 9412055 TI - [The early diagnosis of HIV infection (AIDS)]. PMID- 9412060 TI - [Experience in organizing surgical care during the combat operations in the Chechen Republic]. AB - In the article the final analysis of organization of surgical care during the local military conflict in northern Caucasus was presented. The real possibilities of different medical units such as single medical company and medical squadron of special assignment was described. Also concrete contents of emergency specialized surgical care and its role in improvement of outcomes in patients with combined combat injuries was reflected. Mortality in this patients was decreased from 25.2 to 12.8%. The defects of surgical care were analyzed and the main ways of improvement of surgical care in local military conflict were presented. In the result of using the principle of moving the medical care closer to the wounded in the whole system of consecutive care the mortality was decreased up to 1.3%. PMID- 9412061 TI - [The results of operations to transect the superficial temporal artery in patients with pathology of the posterior portion of the eyeball]. PMID- 9412063 TI - [Hydroxyapol and Kolapol: their use in dental and surgical practice]. PMID- 9412062 TI - [The anti-arrhythmia action of tiracizine in patients with ischemic heart disease and decreased myocardial contractile function]. PMID- 9412064 TI - [The use of Ciprobay in the infectious complications of traumatic illness]. PMID- 9412065 TI - [Terminology in military preventive medicine: its history, status and outlook for improvement]. PMID- 9412066 TI - [Changes in the work capacity of the operators of command-measuring systems during daily duty]. AB - Through 12 hours of work the operators of command-measuring complexes had initial signs of exhaustion, showed them-self by decrease of health state, activity, mood, increase of latent period of simple sensorimotor reaction. These changes of a functional condition had no effect on quality of fulfillment of target problems. At the end of daily duty exhaustion, described by deterioration of health state, increase of operators' anxiousness, rapid pulse, reduction of time of delay of breath, increase of time of instability of sensorimotor reactions, amount of faulty actions, reduction of speed of mental processes and distribution of attention were developed. PMID- 9412067 TI - [The improvement of portable medical equipment]. AB - In article the questions about modernization of portable equipment for rendering the first, medical and the first medical qualified care to the wounded and injured on the battle field and at the stages of medical evacuation are analyzed. The problems of creation of portable equipment for the medical specialists for rendering the urgent care to the injured and patients outside of medical establishments in peace time are considered. Development, carried out by the authors, have allowed to improve equipment of the medical service personnel. PMID- 9412068 TI - [The first evacuation in the Russian Army of the wounded and sick by railroad]. PMID- 9412069 TI - [An outstanding figure in Russian military medicine (Daniil Kirillovich Zabolotnyi)]. PMID- 9412070 TI - [The contribution of Z. M. Volynskii to the development of military medicine]. PMID- 9412071 TI - [Microelements balance and correction in athletes++ under high muscular load]. AB - The balance studies among the high-qualified adult sportsmen during the winter period of practices had shown that of the day of a 30-km cross the contents of iron, copper and manganese in the food ration fell to the lowest level of the physiological standard of people not going into sports due to sufficient physical load the process of microcomponents' secretion through bowels and kidneys was outstripping their replenishment from food. The balance of all three microcomponents was negative. During the following three days after the cross due to disbalanced food ration caused by the content of microcomponents the losses of iron and copper were not compensated. The enrichment of the food rations by the set of components caused the hold-up of iron, copper and manganese in sportsmen bodies. The increased usage of the medical iron resulted in sufficient growth of copper and manganese excretion through alimentary canal. PMID- 9412072 TI - [Inhibiton of endogenous synthesis of nitroso compounds by Selenium in rats]. AB - The inhibiting effect of organic Se (selen-enriched yeast Bioselen) on the endogenous synthesis of N-nitrosubstances was investigated in the Wistar rats, receiving 15 mg of sodium nitritis and 24 mg of diethylamin per 1 kg of bodyweight during 22 days. The level of nitrosoprolin synthesis and (NPro) and the level of nitrosodiethyl (NDie) in the stomach of rats served as the main indices. The highest level of NPro and NDie were revealed in the rats, without selen supplementation (581.2 +/- 113.3 mg per kg of bodyweight and 29.8 +/- 3.0 mg per kg of bodyweight). The highest inhibiting effect of Se was 54.5% for NPro and 54.7% for NDie and it was shown for the Se concentration of 1.5 mg per 1 kg of forage. The increase of Se dosage to 3.0 mg per 1 kg of forage was less effective and resulted in 25.5% of inhibiting of NPro u 47.0% - NDie. PMID- 9412073 TI - [Content of ceruloplasmin as a source of copper in breast milk at different stages of lactation]. AB - The concentration of ceruloplasmin decreased about 10 times during the first 20 days of lactation, judging from both immunoelectrophoretic quantitation and oxidase activity measurements. On the other hand, the daily copper consumption per 1 kg of newborn body weight remains constant (about 10 micrograms per day). The artificial fed children receive many folds excesses of the copper ions. The mechanisms, controlling the consumption of copper by newborns via the modulation of ceruloplasmin gene expression in the cells of the lactating mammary gland in the mother are discussed. PMID- 9412074 TI - [Nutritional status of early age children in Kazakhstan]. AB - An assessment of nutritional status children under 3-years old was given, which conducted within Kazakstan Dimography Health Survey, 1995. 717 children under 3 years old were examined, including Kazak--421, Russian--161 and other ethnicities -135. An assessment of nutritional status was conducted by the anthropometric indices: Height/Age, Weight/Height, Weight/Age. The most expressed degree of undernutrition was determined among rural children, Kazaks, and children living at the South and Center regions of republic. Demographic and background characteristics are the main determinants for nutritional status of children. Birth parity, birth interval, mother's educational level have influence for nutritional status of children. PMID- 9412076 TI - [Diet characteristics of Far North region population depending on the immune system status]. PMID- 9412075 TI - [Prophylactic food products in a diet of patients with diabetes++ mellitus, type II]. AB - Usage of special prophylactic products, such as low caloric vitamins and b carotene enriched soft drink "Gold Ball", b-carotene oileous solution and salt with low sodium content in patients with insulin independent diabetus resulted in better vitamins and b-carotene provision, decrease of the incidence of hypovitaminosis, improvement of antioxidant status, and was accompanied by the decrease of arterial blood pressure. These data allowed us to recommend wide usage of these products in the diet of patients. PMID- 9412077 TI - [Lipid peroxidation in patients with ischemic heart disease, hyperlipidemia and/or hypertension while polyunsaturated omega-3 fatty acids of plant and animal origin were in their diet]. AB - Usage of antisclerotic diet with PUFA n-3 from vegetable or animal source in 326 patients with IHD, HL and HBP resulted in positive dynamic of clinical manifestation, blood lipids and coagulogramms of the patients. The favorable changes of membrane lipids and the decrease of intensity of lipid peroxidation was also revealed. The decrease of primary and secondary metabolites of lipid peroxidation could be a result of hypolipidemic effect of the diets and sufficient level of vitamin E. We suppose that vitamin A and b-carotene content of butter "Laplandia" had additional mild antioxidant influence. PMID- 9412078 TI - [Effect of soluble dietary fibers on cholesterol metabolism in hyperlipidemic patients]. AB - Therapeutic effectivity of new extrusion products "Jantar" have been studied. These products include 10% of dietary fiber gumarabic. It was shown that consumption of products "Jantar" caused a 18.1, 23.2, 16 and 16.3% decrease in the levels of total cholesterol, LDL-cholesterol, tryglicerols and apoprotein B respectively. A decreasing body mass index and hypoglycamic effect also have been observed. Thus, extrusion products "Jantar" may be recommended for diet therapy of hypercholesterolemia, obesity and diabetus melitus. PMID- 9412080 TI - [Once again on food quality]. PMID- 9412079 TI - [Diet therapy with soy proteins for chronic stomach ulcers]. PMID- 9412081 TI - [ Indicators of thiamine content and energy homeostasis in alcoholic psychosis]. AB - In dynamics of acute alcoholic psychoses--delirium and hallucinosis-- considerable deficit of thiamine in the organisms of ill people that can not be compensated while desintoxicative treatment and signs of energy homeostasis changes and of deterioration of tissues reserves depending on the type of psychoses were determined. Pathogenic peculiarities of the revealed disorders were analysed, possible compensative mechanisms connected in particular with the rise of pyruvate utilization speed and activation of glyconeogenesis while leaving the psychotic state were considered. PMID- 9412082 TI - [Vitamins and immune system: vitamin E]. PMID- 9412083 TI - [Toxoplasmosis screening: tu felix Austria?]. PMID- 9412084 TI - [Chronic peritoneal dialysis as home therapy in childhood--risks and complications]. AB - BACKGROUND: The aim of our study was to ascertain the complications of chronic peritoneal home dialysis in childhood. PATIENTS: 17 children were treated by ambulatory peritoneal home dialysis between 1984 and 1994 at the paediatric dialysis unit of the University Children's Hospital in Vienna, Austria. Their average age was 6.5 years (1 week to 12 years); 7 (41.2%) children were below school age (< 6 years). RESULTS: In our observation period of 369 dialysis months (DM), the average duration of dialysis was 21.7 months (4.0-74.3). In relation to total DM the incidence of peritonitis was 1:23.1 of exit site infection 1:14.8 and of catheter related complications 1:41.0. 5 children developed hernias. 5 children were switched to haemodialysis and 8 children received kidney transplants. 2 children died from non-dialysis-associated causes. CONCLUSION: Peritoneal dialysis, in contrast to haemodialysis, is a home treatment modality applicable even to infants. The most common complication is infection. Our data and the European and North American literature show that by close ambulatory monitoring and special hygenic procedures peritonitis frequency can be markedly reduced. PMID- 9412085 TI - [Toxoplasmosis diagnosis in pregnancy: computer assisted follow-up interpretation of serologic tests]. AB - Primary infection with Toxoplasma gondii during pregnancy can result in fetal infection with serious sequelae for the unborn if not treated properly. Early diagnosis enables drug therapy and significantly reduces the risk of fetal disease. A systematic serological screening procedure was established in Austria in 1975 to detect primary toxoplasma infection as early as possible during pregnancy. Since the screening program is based solely on observation and interpretation of serological data, the question arises whether a knowledge-based system for automatic interpretation can achieve a sufficient interpretative accuracy for introduction to routine work. For this reason the system Toxopert-I was developed. The system is aimed at facilitating routine laboratory work, as well as assuring quality by setting standards for therapy. The required knowledge base was designed as a knowledge graph, each state representing a certain interpretation. One or more available serological test results cause the knowledge graph to change its current state. If all available test results are processed, the final state reached corresponds to the respective current interpretation for the patient. A retrospective analysis of 1000 pregnant women yielded a total diagnostic sensitivity and specificity of over 99% in comparison with the clinician's diagnosis which was used as the Gold Standard. PMID- 9412086 TI - [Urgent diagnostic steps in acute manifestation of intraspinal processes]. AB - Immediate diagnostic clarification is required in patients who develop acute or subacute symptoms suggestive of an intraspinal lesion. In case of symptoms indicating a monoradicular lesion a spinal CT investigation is mostly sufficient. Since polyradicular syndromes are often due to inflammation, examination of the cerebrospinal fluid is the most important diagnostic measure. However, in case of symptoms suggestive of intramedullary lesions, spinal MRT is by far the most effective diagnostic procedure. In patients with symptoms suggestive of a lesion of the cauda equina spinal CT is sufficient in most cases as a first measure, particularly if the lesion can be precisely localized by clinical examination. The decision as to which diagnostic method should be performed first is relevant mainly because of the limited availability of MRT examinations within the daily clinical routine. MRT should thus be used selectively in patients with lesions that cannot be identified by alternative diagnostic methods. PMID- 9412087 TI - [Imaginary letters to patients with diabetic nephropathy or does overview come from oversight?]. PMID- 9412088 TI - Nutrition and Fitness: Evolutionary Aspects, Children's Health, Programs and Policies. Proceedings of the 3rd International Conference. Athens, Greece, May 24 27, 1996. PMID- 9412089 TI - [The scent marking of territory of gerbils: a comparative analysis exemplified by 4 species of the genus Meriones]. AB - Sex-age and seasonal variability of the ventral glands and different stereotypes of scent marking behaviour in four Meriones species (M. unguiculatus, M. meridianus, M. libycus, M. tamariscinus) have been studied in nature and under semi-natural conditions. Two major ways of olfactory marking are considered: by secretion of the ventral glands and by "signal heaps" with urine and feces. Intraspecific and inter-species variability of marking activity is investigated. The ventral glands start to function at the period of preparation of a generative system to reproduction. The peak of secretary activity of gland and maximum of two types of marking activity is observed in spring and early summer, i.e. the period of active reproduction. The maximum of two types of the marking activity is observed during this period. In M. tamariscinus and M. meridianus the marking by the ventral gland is prevailing mode of the territory scent marking, while Mongolian gerbils (M. unguiculatus) prefer to use "signal heaps" Libyan gerbils (M. libycus) in this relation take an intermediate position. At the non productive period a level of marking activity is on 10-20 times lower than at the reproductive season. Besides hormonal, social factors were also important for regulation of marking activity. By influence of these factors the differences in the level of marking activity in high-rank and low-rank individuals and differences in patterns of a spatial distribution of scent marks in individuals of different hierarchical rank is explained. Functional significance of various ways of territory scent marking is discussed. PMID- 9412090 TI - [Indications and results of para-sternal mediastinoscopy in tumors of the anterior mediastinum]. AB - From January 1990 to December 1995 79 patients with mediastinal lesions were seen for parasternal mediastinoscopy at the Ruhrlandklinik Essen. Diagnosis was achieved in 91.1%. In 7 cases (8.9%) the diagnosis was not established. Six intraoperative complications occurred following mediastinoscopy: minor bleeding in five instances from the internal mammary vessels and one significant bleeding by injuring the V. cava. Postoperative minor wound infections occurred in five patients. Two pneumothoraces had to be managed by chest tube drainage. One patient with metastatic lung cancer died of respiratory failure. PMID- 9412092 TI - [Efficacy of minimally invasive thoracic surgery in diagnosis of pulmonary coin lesion]. AB - AIM OF THE STUDY: With only a few exceptions every pulmonary nodule of unknown dignity has to be clarified by biopsy. Aim of the present study was to analyze the usefulness of minimal invasive thoracic surgery in patients with indeterminate pulmonary lesions. METHODS: In the study included were 121 patients (67 male/54 female), who were primarily treated by minimal-invasive surgery during the period 1992-1995. Preoperatively, a single pulmonary nodule was diagnosed in 89 patients. 32 patients had two or more lesions. RESULTS: Using the minimal invasive approach a pulmonary nodule was successfully localized and resected in 83 (68.5%) patients. In 33 patients the operation was classified as diagnostic, in 36 patients benign or inflammatory tumors were completely resected and in 14 patients palliative resections of malignant tumors were performed. Overall, an extension of the operative approach was necessary in 38 (31.5%) of the patients. In 23 patients a "mini-thoracotomy" (< 5 cm) was used to localize the nodule. A standard thoracotomy was performed in 15 patients, mostly because of massive adhesions. CONCLUSIONS: The minimal invasive approach has become a routine procedure in thoracic surgery and is extremely useful in the diagnosis of indeterminate pulmonary nodules. PMID- 9412091 TI - [Assessment of current pleurodesis procedures exemplified by pneumothorax]. AB - This survey addressed common methods of video-assisted thoracoscopic pleurodesis for spontaneous pneumothorax. A questionnaire asking for frequency, recurrence rate and complications of the different methods of pleurodesis was sent to all hospitals that belong to the German Society for Thoracic Surgery. 19 hospitals reported on a total of 1365 operations. 88 recurrences (6.5%), 26 severe bleeding complications (1.9%), 39 persisting air leaks (2.9%) and two hospital deaths (0.1%) had been observed. Pleurectomy and pleural abrasion were the most common procedures but induced significant (p = 0.01) more bleeding complications (3.1% and 2.6%) than all other methods of pleurodesis (0.4%). Overall recurrence rates depended significantly on the chosen procedure (p = 0.0013). Pleurectomy (4.4%) and coagulation of the pleura (2.7%) showed better results than the average. Due to smaller numbers of operations and the widely differing results this significance cannot be shown for the individual recurrence rates of the different clinics. This survey demonstrated a trend towards lower rates of recurrence and complications after coagulation of the pleura parietalis. The retrospective character of the investigation and extremely different recurrence rates for different hospitals demand cautious interpretation of these results. PMID- 9412093 TI - [Value of intraoperative cytology in thoracic surgery]. AB - Cytological investigations are established in thoracic surgery beside intraoperative histological examinations. We use these intraoperative cytological investigations as an additional diagnostic option for surgical decisions at our hospital for more than 15 years. 1008 intraoperative cytological findings were compared with the final histological results in a retrospective study to investigate the security of these cytological findings. The diagnostical sensitivity for the detection of any malignancy was 97.7%. A correlation of cytological and histological results was found for benign tumors in 96.2% and for inflammatory diseases in 89.2%. The cytological and histological diagnoses regarding the real histological type of malignant tumors corresponded in only 667 of 834 cases. Due to these findings we conclude that intraoperative cytology is very useful to diagnose malignancy. However, additional diagnostic and clinical parameters have to be used for making final intraoperative decisions. PMID- 9412094 TI - [Recurrence after curative operation of non-small-cell bronchial carcinoma]. AB - The poor prognosis of bronchial carcinoma is reflected among other things in a high recurrence rate. In general, recurrence is inoperable and only suitable for conservative/palliative management. The rate of curative surgical reintervention may be increased by early identification of recurrence. The value of tumour follow-up and the role of reoperation need to be assessed. 150 patients who underwent curative resection of non-small-cell lung cancer were followed-up for a mean of 4.5 years as part of a comprehensive after-care program. Fifty patients (33%) developed a recurrence at a mean of 13 months after operation. A second curative resection was possible in 9 patients (6%) with a mean survival of 22 months. Six of these patients had re-thoracotomy (completion pneumonectomy in 4 for local recurrence, mediastinal metastasectomy in 2), and in 3 patients a solitary cerebral metastasis was excised. Our results show that with focused, schematic tumour follow-up early recognition of recurrence is possible. Despite this, reoperation is only indicated in a selected group of patients because of multifocal recurrence, or local or functional inoperability. Further intensification of tumour follow-up is limited by personnel, logistical, and financial considerations. As an alternative, individualised, function-oriented tumour after-care could be considered. PMID- 9412095 TI - [Surgical aspects of pulmonary thrombendarterectomy]. AB - Pulmonary thromboendarterectomy is an accepted operative procedure for treatment of pulmonary hypertension due to chronic embolism. Despite its proven value this procedure has been established at very few centers worldwide. In this paper we report our actual operative concept and operative results. Between 8'89 and 4'96 127 patients were operated with use of extracorporeal circulation, deep hypothermia and circulatory arrest. After analysis of the initial high perioperative mortality (26%, 29/108) our operative and postoperative concept changed since 11'94: 1. central incision of both pulmonary arteries, 2. endarterectomy exclusively during circulatory arrest, 3. prolonged reperfusion to 37 degrees C, 4. pressure controlled ventilation, NO-inhalation, early extubation, and 5. modified vasopressor therapy. Preoperatively 12 of the 19 patients were in NYHA class III and 6 in class IV. Mean pulmonary artery pressure was 52(17) mmHg with a calculated pulmonary resistance of 1013(579) dynes.s.cm-5. Mean circulatory arrest time was 37 min (19-57 min) (bypass time 345 min, (240 430 min)). Perioperatively two patients (11%) died (multiorgan failure; rethrombosis of pulmonary artery/right heart failure), all other patients survived (89%). Perioperative complications included reversible renal failure, delirium and postcardiotomy syndrome (1/2/1). Mean pulmonary resistance was postoperatively significantly reduced (362(124) dynes.s.cm-5) (p < 0.01). Early results of pulmonary thromboendarterectomy can be improved by consequent modifications of the intra- and postoperative concept. PMID- 9412096 TI - [Thymectomy in myasthenia gravis]. AB - Myasthenia gravis is a relatively uncommon autoimmune disorder of neuromuscular transmission. Surgical therapy plays an important role in addition to medical treatment. Follow-up results of 52 patients with thymectomy are presented. Between 1984-1996 thymectomy via median sternotomy was performed in 52 patients with myasthenia gravis (female = 28, male = 24). The score described by Ossermann and Genkins was used for classification. According to this classification, we found 12 patients in class II(I), 21 in class IIA, 17 in class IIB and 2 in class III, respectively. A thymoma was found in 19, follicular lymphoid hyperplasia in 24 and an atrophic thymus in 9 cases, respectively. There was no mortality. Severe postoperative complications consisted of bleeding and reoperation in one patient and another patient developed a sternal instability with consecutive operative refixation. Follow-up evaluation after a mean period of 36 months (min. 6 months, max. 130 months) revealed a relief of myasthenic symptoms in 37 patients. Thymectomy is effective in the treatment of myasthenia gravis with a low complication rate. PMID- 9412097 TI - [Value of thoracoscopy in thoracic trauma--initial experiences]. AB - The aim of this study was to assess the role of thoracoscopy in the evaluation of the cause of persistent intrathoracic bleeding, air leak, or nuclear basal opacification after blunt thoracic trauma. As a result, a decision to proceed to early thoracotomy could be made, or an attempt of thoracoscopic haemostasis, haematoma evacuation, or fistula closure was possible. Twelve patients (9 male, 3 female, mean age 33,7 years) with blunt thoracic trauma underwent video-assisted thoracoscopy under general anaesthesia with double-lumen endotracheal intubation and one-lung ventilation. The indication for operation was made after assessment of chest X-ray and CT findings, pleural ultrasound, and the volume and quality of pleural drainage. Persistent pneumothorax was shown to be due to traumatic rupture of a bulla in two cases and to parenchymal air-leak from a small lung laceration in two cases, all of which were treated endoscopically. In two cases a diaphragmatic rupture was confirmed as the cause of basal shadowing and in one case a major lower lobe laceration was identified as the cause of a persistent haemopneumothorax. In three cases, a fluid collection which could not be evacuated through a pleural drain was shown to be an organised haematoma and was removed endoscopically. Video-assisted thoracoscopy is helpful in the diagnosis and treatment of thoracic trauma, allowing early recognition of injuries that require thoracotomy. It is indicated for persistent (but not life-threatening) intrathoracic bleeding, unresolving pneumothorax, and unclear basal opacification. Therapeutic parenchymal tissue glue application and suturing as well as local resection and haematoma evacuation can be performed with this technique. PMID- 9412098 TI - [Can diagnosis and subsequent trauma management of the multiple trauma patient with blunt thoracic trauma be improved by early computerized tomography of the thorax?]. AB - OBJECTIVE: The aim of this prospective study was to evaluate, whether early thoracic computed tomography (TCT) is superior to routine chest x-ray (CXR) in the diagnostic work up of blunt thoracic trauma and whether these additional informations influence subsequent therapeutical decisions in the early management of severely injured patients. PATIENTS AND METHODS: In a prospective study of 103 consecutive patients with clinical or radiological signs of chest trauma (94 multiple injured patients with chest trauma, 9 patients with isolated chest trauma) with an average ISS of 30 and an average AIS thorax of 3 initial CXR and TCT were compared after first assessment in our emergency department of a level I trauma center. RESULTS: In 67 patients (65%) TCT detected major complications of chest trauma, that have been missed on CXR [lung contusion (n = 33), pneumothorax (n = 27), residual pneumothorax after chest tube placement (n = 7), hemothorax (n = 21), displaced chest tube (n = 5), diaphragmatic rupture (n = 2), myocardial rupture (n = 1)], in 11 patients only minor additional pathologic findings (dystelectasis, small pleural effusion) were visualized on TCT and in 14 patients CXR and TCT showed the same pathological results. 11 patients had both CXR and TCT without pathological findings. The TCT scan was significantly more effective than routine CXR in detecting lung contusions (p < 0.001), pneumothorax (p < 0.005) and hemothorax (p < 0.05). In 42 patients (41%) the additional TCT findings resulted in a change of therapy: chest tube placement or chest tube correction of pneumothoraces or large hemothoraces (n = 31), change in mode of ventilation and respiratory care (n = 14), influence on the management of fracture stabilization (n = 12), laparotomy in cases of diaphragmatic lacerations (n = 2), bronchoscopy for atelectasis (n = 2), exclusion of aortic rupture (n = 2), endotracheal intubation (n = 1), pericardiocentesis (n = 1). CONCLUSIONS: TCT is highly sensitive in detecting thoracic injuries after blunt chest trauma and is superior to routine CXR in visualizing lung contusions, pneumo- and hemothorax. Early TCT influences therapeutic management in a considerable subset of patients. We therefore recommend TCT in the primary diagnostic work up of multiple injured patients with suspected chest trauma, because early and exact diagnosis of all thoracic injuries along with sufficient therapeutic consequences may reduce complications and improve outcome of severely injured patients with blunt chest trauma. PMID- 9412100 TI - [Changes in therapeutic principles in fractures of the extremities with severe soft tissue injuries exemplified by tibial fracture]. AB - Functional results after open fractures have been improved during the last decades. Especially the rates of amputation and chronic osteitis after open tibial fractures have been reduced from 30% to less than 5%. The initial management of this type of fracture includes reconstruction of the perfusion of the involved vessels, subsequent debridement with resection of avascular tissues, decompression of compartments by fasciotomy and initial shortening of the tibia by osteotomy and followed by callus distraction in order to achieve the physiological length of the leg. Cortical bone with periostal stripping has to be covered by local muscle transfer or by free vascularized tissue transfer within 3 7 days. Bone defects are either reconstructed by cancellous bone graft or, if the defect is longer than 2 cm, by continuous segmental transfer, according to the technique described by Ilizarov. PMID- 9412099 TI - [Tracheobronchial injuries in blunt thoracic trauma]. AB - Tracheobronchial lesions after blunt chest injury are seldom (0,5-0,7%). Diagnostic and therapeutic strategies in 16 own cases and a review of the literature are presented. Own experiences: Locations of the lesions were main bronchus (10), bronchus intermedius (2), and trachea (4). Rupture was total in five cases, and partial in seven. In four patients the mucosa only was ruptured. Initial symptoms: Subcutaneous emphysema (13), pneumothorax (9), respiratory insufficiency (5), lung lesion (5), but tracheal bleeding in five cases only. Diagnosis mainly by bronchoscopy (8 early, 4 late), but in 4 cases after thoracotomy. TREATMENT: In cases of total rupture, there were three anastomoses of the bronchus and one of the trachea, and one pneumonectomy. In all partial ruptures, there was suturing of the lesion. Mucosa lesions were treated conservatively. RESULTS: 1 empyema, 2 ex. leth. (bilateral pneumonia 7.d., multiple organ failure 20. d). FOLLOW UP: 9 patients free of symptoms, 5 patients with respiratory problems. The symptoms "mediastinal emphysema and continuous air leakage through the chest tube or persistent atelectasis of the lung" are indications for urgent bronchoscopy and early surgery. Long-term results are good in 70%-90% of the cases. Not diagnosed lesions can result in tracheobronchial stenosis and infections of the lung later on, to be treated by lung resection only. Total bronchial ruptures can result in strictures and non-infected atelectasis, resection of the stricture and reinflation of the lung being successful in 60%-70% of these late diagnosed cases. PMID- 9412101 TI - [Diagnostic and therapeutic standard in meniscus lesions--results of a survey]. AB - With a survey among surgeons and orthopaedic surgeons in German speaking countries the presently applied diagnostic and therapeutic schemes of meniscus lesions were investigated. In 322 questionnaires altogether 43.958 meniscus lesions were reported. In over 90% arthroscopy serves as diagnostic and therapeutic tool. Two thirds of the surgeons suture a longitudinal tear of the outer zone of the meniscus, all the other meniscus lesions were foremost treated by partial resection. The postoperative treatment after meniscus surgery is very inconsistent. Altogether less preserving meniscus operations were performed than it would be possible according to the literature. On one hand new therapy regimens are acknowledged by clinical practice with considerable delay, on the other hand the importance of menisci for the integrity of the knee-joint necessitates a further improvement of reconstructive techniques for preservation of the menisci. PMID- 9412102 TI - [Transplantation ordinance--establishing brain death criteria, revised informed consent and much more]. PMID- 9412103 TI - [Percutaneous laparoscopic gastrostomy]. AB - Percutaneous endoscopic gastrostomy (PEG) has now largely replaced the conventional operative procedure of gastrostomy. However, in patients with endoscopically non-passable pharyngo-oesophageal tumours or facial injuries rendering the stomach inaccessible to endoscopy, this technique has so far been unavailable. In this group of patients it is nevertheless possible to apply minimally invasive percutaneous gastrostomy using laparoscopy (PLG). The employment of a purse-string suture clamp facilitates the performance of the procedure and makes it reliable. PMID- 9412104 TI - [From the history of surgical instruments: 6. Sauerbruch's instruments for thoracic surgery]. AB - The historical development of thoracic surgery, which was established in Germany in the years 1904/1905 and the armamentarium of the chest designed by Ferdinand Sauerbruch (1875-1951) are described. Sauerbruch designed between 1904 and 1920 many instruments to facilitate thoracic surgery, e.g. several rib retractors, a special rib shear for the first rib, a rib shear for all ribs other than the first. Furthermore he designed a special lung grasping forceps with rubber covered jaws, a lung compression forceps and a fenestrated lung retractor to facilitate lung resections. PMID- 9412105 TI - [The effect of optical radiation on the migration of third stage larvae of Oesophagostomum quadrispinulatum out of feces]. AB - The influence of optical radiation on the emigration of third stage larvae out of faeces was investigated by exposing faeces containing infective larvae of Oesophagostomum quadrispinulatum to broad-band radiation of a sun-simulating wavelength spectrum corresponding to a sunny day, a cloudy day, dawn, and a full moon night, as well as to monochromatic radiation of different wavelength spectrum at dawn and to complete darkness. It was demonstrated that third stage larvae of O. quadrispinulatum were able to differentiate between daylight and darkness responding to different irradiances with very high emigration rates at irradiances corresponding to dawn, and significantly lower emigration rates corresponding to full-moon light, and darkness as well as a cloudy and sunny day. Infective larvae reacted to monochromatic radiation of different wave-length spectrum at dawn and showed significantly higher emigration rates at the violet, green, yellow, and red light wavelength compared to darkness. PMID- 9412106 TI - [Defensins and antibiotic peptides related to them in the evolution of animal protective systems]. PMID- 9412107 TI - [The intensity of the peroxidation of lipids and their fatty acid composition in Drosophila melanogaster strains differing in their adaptive value]. PMID- 9412108 TI - [Changes in hemoglobin affinity for oxygen in sheep under the influence of ethanol]. PMID- 9412109 TI - [A comparative analysis of the EEG spectral components at rest and in the active state in feline ontogeny]. PMID- 9412110 TI - [A comparative study of the interaction of cholinesterases in Pacific Ocean squid (Berryteuthis magister) and some mammals with silatrane-onium inhibitors]. PMID- 9412111 TI - [Brain insulin and its receptors in the natural fasting of fishes and cyclostomes]. PMID- 9412112 TI - [A theoretical analysis of the primary structure of the SEC59 yeast gene product. The evolutionary relationship to enzymes of the dolichol cycle and channel proteins]. PMID- 9412113 TI - [The hormonal function of the adrenals and gonads in man and monkeys during aging]. PMID- 9412114 TI - [The cortisol content in the fetal adrenals of the silver fox Vulpes fulvus during selection for the domesticated type of behavior]. PMID- 9412115 TI - [The ontogenetic characteristics of a disorder in the perception of different emotions in speech under the influence of noise]. PMID- 9412117 TI - [The evolution of the support systems of cellular homeostasis--the basis of the progressive evolution of organisms]. PMID- 9412116 TI - [The evolution of the functions of inorganic polyphosphates at different stages in the phylogenetic development of living creatures]. PMID- 9412118 TI - [The probable substrate of subjective states in man and the development of this substrate in evolution]. PMID- 9412119 TI - [The advent of a new class of antihypertensive agents: angiotensin II receptor antagonists]. AB - The objective of this article is to give more information about the pharmacology of and recent clinical data on the angiotensin II receptors antagonists. The angiotensin II receptors antagonists, of which Losartan will be the first representative on the Belgian market, constitute a new therapeutic class in the treatment of hypertension and even heart failure. They are non peptic and orally active and their long mechanism of action allows one daily administration to improve therapeutic compliance. These agents block all known effects of the angiotensin II through binding to the AT1 receptors. Thanks to this unique mechanism of action they reduce blood pressure with a lower incidence of the adverse effects commonly associated with other antihypertensives. In controlled clinical trials, overall incidence of adverse experiences was comparable to placebo. Addition of thiazide-type diuretics provides additive efficacy. PMID- 9412120 TI - [Unexpected origin of recurrent ascites: apropos of 2 cases]. AB - The authors report the cases of two patients suffering from ascites attributed for several years to a non pericarditic aetiology. The first patient presented with a diagnosis of right cardiac failure secondary to a right myocardial infarction. Cardiac catheterisation, magnetic resonance imaging and transoesophageal echocardiogram allowed to establish the diagnosis. In the second case, ascites was attributed to cirrhosis. Presence of pericardial calcifications, visible on a chest X-Ray led to suspect constrictive pericarditis. In both cases, ascites contained a high protein level. A pericardectomy allowed a favourable outcome in both cases. Thus, a diagnosis of constrictive pericarditis must be evoked in face of ascites of unclear origin and a normal cardiac size. PMID- 9412121 TI - [Treatment and glycemic control of 1200 diabetic patients enrolled at the National Institute of Disease-Disability Insurance diabetes convention]. AB - We have studied the characteristics of insulin therapy and home blood glucose monitoring of 1.200 diabetic patients (590 males, 610 females), mainly type I adults (age: 43 +/- 19 years, mean +/- 1 SD), attending a licensed diabetes center in Belgium which benefits from a National Health Service convention system for providing education and home blood glucose monitoring kits and disposables to diabetic subjects. 50% of these patients were treated with 2 daily insulin injections, while 22% were treated with 3 and 24% with 4 daily injections. 4% of the subjects were treated with a continuous subcutaneous insulin infusion. Patients treated with four daily injections were younger than those treated with other insulin schemes (p < 0.001). Home blood glucose monitoring strip consumption was 2.2 patient-1 day-1. The mean HbA1C level was 8.63 +/- 1.55% [8.54 +/- 1.46 in males and 8.72 +/- 1.62% in females (NS)]. In more than one third of the subjects treated with injections, HbA1C was lower than 8% HbA1C was also below 8% in 50% of patients using 1100-1200 strips year-1. PMID- 9412122 TI - [Liver histopathological characteristics in patients with HCV-positive sera]. AB - In order to have knowledge on the histopathologic characteristics of the HCV infections in our geographical area and its relation with some epidemiologic variables, a series of 54 biopsies of Anti HCV (R) patients was analyzed. The histologic lesions found in this study correspond mostly to patients with relatively early infections, on the contrary to other studies of the some kind. The most frequent histopathologic diagnosis were chronic hepatitis 38/54 (70.4%), steatosis 4/54 (7.4%) and 12/54 (22.2%) with no changes. The presence of lymphoid follicles in the portal tracts was the most frequent histological change in this series (66.7%), followed by the alteration in bile ducts (53.7%); they occurred in a significantly higher proportion in the biopsies which had a diagnosis of chronic hepatitis (p = 0.02) (p = 0.000002). The presence of steatosis and acidophilic bodies in the acinus were found in nearly one third of the biopsies. This findings suggest that the hepatic damage in the anti HCV reactive patients might take place through immunologic mechanisms and cytopathic direct action. It was not found that histologic changes produced by HCV might differ according to epidemiologic variables (post-transfusional, drug abuse i.v. and sporadic). PMID- 9412123 TI - [Systematic study of gastrectomy specimens for cancer, in search of synchronous carcinomas]. AB - Multiple carcinomas were searched in 50 successive gastrectomies, 17 females (34%) 33 males (66%) from 40 to 83 years old. Macroscopic handling of the specimens included pinning on a board, fresh, half-fixed and completely fixed examination, not only by naked eye but also through a magnifying glass. The stomach were totally studied, according to Japanese method. A satisfactory slide was obtained from each block, stained with H.E. and examined by each of the authors. Synchronous tumors were found in 4 cases (8%), 3 males and one female. Compared to the main lesion, two cases showed a proximal tumor, one case showed a distal tumor, and the last one showed two tumors, one of them proximal and the other distal. Three tumors were macroscopically and two were microscopically detected. Two of them coexisted with advanced carcinoma and three with early carcinoma. Four of them were located in mucosa with intestinal metaplasia. Histologically, four cases belong to the same type of the original tumor, and one of them did not coincide. We conclude that stomachs resected for cancer must be thoroughly and methodically studied before and after fixation. We suggest that resection must include the whole mucosa with intestinal metaplasia, in order to avoid leaving a synchronous tumor in the gastric stump. PMID- 9412124 TI - [Microcystic serous cystadenoma of pancreas. Clinicopathological and surgical features]. AB - The microcystic serous cystadenoma of pancreas or glycogen "rich" cystadenoma is a rare entity. We studied five case of this cystadenoma in adult patients ages 47 68 (58 was the mean), four of which were women (80%). The clinical presentation was varied. There was a prevalence of expansive manifestations with epigastric pain in three patients, and extrahepatic bile duct obstruction in other two. A distal tumour was revealed by the diagnostic methodology used (ultrasound and TAC) in three patients, and cephalic tumour in two, with a mean size of 8.8 cm. in diameter. A distal pancreatectomy was performed in two patients, a cephalic pancreatoduodenectomy was performed in one in relation with the presence of extrahepatic bile duct carcinoma, and the other two were treated with a partial cephalic pancreatectomy (enucleation). The nosological diagnose was post-surgical in all case of study. A prognosis for every case was dependent of the associated pathology. PMID- 9412126 TI - [Role of liver biopsy in hepatitis C]. PMID- 9412125 TI - [Preventive role of colonic polypectomy]. PMID- 9412127 TI - [Hepatoportal sclerosis. 10 years of experience at the National Institute of Pediatrics]. AB - OBJECTIVE: To assess the frequency and clinical picture of Hepatoportal Sclerosis in a population of Mexican children of the Instituto Nacional de Pediatria, Mexico City. BACKGROUND: Hepatoportal Sclerosis is a disease of unknown etiology. It's diagnosis is difficult. The main clinical presentation is splenomegaly with or without hematemesis (portal hypertension). Splenoportography and liver histology study are the best procedures for diagnosis and must be performed by experts. METHODS: We studied 7/106 children with portal hypertension during a period of 10 years, who were seen at the Instituto Nacional de Pediatria, Mexico city. Inclusion criteria were specific findings of splenoportography and histologic changes in liver biopsy. RESULTS: We found 7/106 children. The main clinical manifestation were splenomegaly and hematemesis. We did not find any previous history of contact with arsenic, vinyl chloride or copper sulfate. In 6/7 children a porto-systemic shunt was performed. Only one received propranolol and sclerotherapy. At the time of this report all children have shown a good clinical course. PMID- 9412128 TI - [Abnormalities in gastric mechanic sensitivity, gastric emptying and electrogastrography in non organic dyspepsia (NOD)]. AB - The aim was to study the alterations in mechanosensitivity, gastric emptying, and electrogastrography (E.G.G.) in a population of patients suffering from N.O.D. eighteen controls (9 males, 9 females, mean age 49.33 years old < SEM 3.62, range 24-74) and 32 patients with N.O.D. (22 males, 21 females, mean age 55.72 years old, SEM 2.87, range 17-86) were studied. Gastric mechano-sensitivity with a latex balloon of low compliance inflated "in phasic" was investigated, and intra balloon pressure was recorded. Gastric emptying with a mixed meal marked with 99 Tc in the solid phase, containing 250 Cal, was studied. E.G.G. was studied using two skin surface electrodes Ag-2C1Ag placed on epigastric area following a probalistic antral axe. Only dominant frequency in each block was considered, and % of total abnormalities on total recording time lesser than 2 c.p.m or more than 4 c.p.m. was considered. Recordings were taken during fast time during 30 minutes, and 30 minutes after a meal containing 250 Cal. Analysis with F.F.T, and spectral running. RESULTS: In 67.92% a delay in gastric emptying was observed. 56.3% did not complete 700 ml. of balloon inflation because of pain, Vs 16.8% in controls (p < 0.001) The slopes of intra-balloon pressure were not different in both groups. (Variance, F-NS). Mean E.C.A was 2.99 c.p.m in control, Vs 3.46 in fasting and 3.64 in postprandial period in N.O.D. (p = NS) Differences in fasting and postprandial % of arrhythmias total time recording were significant in N.O.D. ("t". 0.02 > p 0.01). Twenty percent of controls showed isolated tachygastria, but dominant frequencies never were higher than 6 c.p.m and never last more than 8% of the total recording time. Sixty eight point seventy five percent of N.O.D. showed arrhythmias. 48% of tachygastrias were in the range 30-60% of total recording time. No differences in gastric emptying between patient presenting pain with = < 700 ml. and < 700 ml. of balloon inflation were seen. Patients with sensorial threshold = < 700 ml. showed less frequent tachygastria (0.01 < p < 0.01). Patients with delayed gastric emptying showed more frequent tachygastria (94.7%, 0.05 < p < 0.02) CONCLUSIONS: a) E.G.G. abnormalities would more frequent in "motor" subpopulation; b) No association between abnormalities in gastric emptying, abnormal thresholds in mechanosensitivity and/or E.G.G. and any clinical subtype of Dyspepsia can be showed; h) although gastric arrhythmia was more frequent in motor than in sensorial abnormalities, it may represents a more generalized disturbance in central modulation in afferents and efferents inputs and outputs. PMID- 9412129 TI - [Sodium saccharin effect on the mice large intestine]. AB - Sodium saccharin has found to be a tumoral promoter in the rat's bladder epithelium, property not demonstrated in humans. Nevertheless, at present there's no references on the possible alterations produced by sodium saccharin in the epithelium of the mice colon. In this work we describe the alterations produced by low doses of sodium saccharin in the epithelium of the mice colon. The changer produced by sodium saccharin consist in pleomorphic microvill with variations of form, length diameter and curvature and demonstrate by transmission electron microscopy. PMID- 9412130 TI - [Parasitosis in an adult population with chronic gastrointestinal disorders]. AB - We worked with 185 middle-class patients above 18 years of age, both sexes, who presented diarrhea and/or chronic gastrointestinal disorders. The faeces were collected serially in formol 10% and processed in the following way: direct microscopy, with and without wet staining, concentration by Ritchie's method, 1% safranine technique for a specific investigation of Cryptosporidium sp., and faecal sieving macroparasites. Twenty eight point six of the studied patients showed at least one enteroparasite in their faeces, 48 harboured one parasite and 5 harboured two parasites. The following parasites were found and their corresponding percentages in the entire studied population are given below: Blastocystis hominis 15.7%, Giardia lamblia 7.5%, Cryptosporidium sp. 1.6%, Entamoeba coli 3.3%, Chilomastix mesnilii 1.1%, Ancylostoma duodenale-Necator americanus 0.5%, Ascaris lumbricoides 0.5%, Enterobious vermicularis 0.5% y Endolimax nana 0.5%. The most frequently found enteroparasites in the positive patients were B. hominis and G. lamblia. Cryptosporidium sp. was diagnosed in only three patients. The source of infection could be presumed in all of them. The symptomatology coincided with that described for this coccid in the bibliography. In spite of the fact that they were HIV seronegative patients the diarrhea was not self-limiting, but the immunologic profile of their relatives remained unknown and no other cause of immunosuppression could be detected with justified chronicity. The treatment with spiramycin was effective. Giardiasis was found in 17 patients, and the source of infection could not be inferred in any of them. They all had chronic diarrhea and their most frequent symptoms were abdominal pain, metallic taste, flatulency and nausea. Most of these patients were harboured one parasite, and only 2 of them simultaneously presented another faecal parasite associated to G. lamblia. Treatment with metronidazole was successful in all of them. Twenty nine patients were found to have B. hominis. The source of infection could not be inferred, this amoeboid was present as the only parasite in 25 patients. Predominant symptoms were flatulence, abdominal distention and colis. All patients suffered from chronic diarrhea, alternating, in some cases, with constipation. Good therapeutic results were obtained with metronidazole. Considering that one third of the patients examined presented faecal parasites associated to chronic disorders, it is important to insist on the detection of parasites to chronic disorders, it is important to insist on the detection of parasites using appropriate diagnostic techniques since the application of specific therapy made their eradication possible as well as relieving the patients' symptomatology. PMID- 9412131 TI - [Effect of microcrystalline cellulose on alkaline gastritis due to bile reflux]. AB - Biliary acids present in gastric content due to a duodena-gastric reflux have been blamed for attacking the gastric mucosa through detergent action on the defensive barrier. This harmful action produces an inflammatory lesion on this mucosa, resulting in a chronic gastritis which is clinically expressed by epigastric pain, heartburn and vomits of biliar aspect among other symptoms, and it has constituted a clinical entity named Alcaline Gastritis by Duodenal-gastric Reflux, more recently, Reactive Gastritis according to the Sydney System. Among dietetic fibers, cellulose has been reported in recent investigations as most active in capturing and inactivating biliary acids: this constitutes a cytoprotective action on the stomach mucosa of patients who suffer this pathology. Concerning these aspects, we studied 50 patients with alcaline gastritis, excluding those who have H. pylori. The patients were diagnosed by endoscopy studies, dosage of total biliary acids in gastric content and biopsy and were divided into two groups of 25 subjects each. The first group (A) received powdered corn starch and the second group (B) powdered microcrystalline cellulose dosed 5 g/day during three months. The results showed that total biliary acids in gastric content decreased at the end of treatment mostly in patients treated with microcellulose, even though there was no statistical significance. From the clinical view point of vue, there was a highly satisfactory response in pain, vomits and heartburn after microcellulose treatment. The endoscopic inflammatory located in the antral region and diffusely, in the whole gastric mucosa, quantitatively improved and there was a significant difference in the antral location. The histological findings at the end of the treatment in either group showed no evolutive variation in the comparative study of the lesions found at the beginning of the treatment. Results are shown in tables. PMID- 9412132 TI - [Acute low digestive hemorrhage in testicular choriocarcinoma]. AB - We report a case in which acute intestinal hemorrhage was the initial manifestation in the evolution of choriocarcinoma of the testis. The diagnosis of bleeding was difficult and required laparotomy. Metastatic lesion of the small bowel should be considered a possible cause of intestinal bleeding in patients with testicular choriocarcinoma. PMID- 9412133 TI - [Ultrasonography findings in eosinophilic gastroenteritis]. PMID- 9412134 TI - [Granular cell tumor of the esophagus. Report of a case and review of the literature]. AB - Granular cell tumor of the esophagus, also called Abrikossof's tumor, is a rare and mostly benign neoplasm which is associated with cancer in 11% of cases reported in the literature. We report herein a 48 year-old-man with granular tumor of the esophagus and carcinoma of the papilla of Vater. it is a unreported association. PMID- 9412135 TI - [Hepatolithiasis: is it a pathology just from the Eastern World?]. PMID- 9412136 TI - [Current approach to intestinal parasitosis]. PMID- 9412137 TI - [Prevalence of antibodies against hepatitis A virus (HAV) in a population of Mexican children]. AB - We studied by ELISA test 450 healthy children looking for IgG and IgM antibodies against virus A hepatitis. 376/450 (86.3%) showed IgG antibodies. 50% of children under one year were positive. At 3 years of age 80% of children from this study were with antibodies IgG and 96% of children at 10 years of age were positive. Only 9 children showed antibodies IgM. PMID- 9412138 TI - [Classic strategies for diagnosis of cryptosporidiosis in patients with AIDS and chronic diarrhea]. AB - Cryptosporidium sp., a protozoa organism, has been increasingly recognized in association with severe enteritis in patients with the Acquired Immunodeficiency Syndrome. The studied subjects included 84 adult patients with AIDS and chronic diarrhea. We describe 14 patients with intestinal infection caused by Cryptosporidium sp. The mean CD4 count in these patients was < or = 300 cells/mm3 (7 out of 14). Examination of duodenal aspirates and feces included dimethylsulfoxide, auramine and acid-fast preparation of concentrated samples. We carried out videoesophagogastroduodenoscopy (VEDA) to visually inspect the mucosa and obtain biopsy specimens. VEDA revealed granular duodenum in ten patients and jasper duodenum in one of them. Duodenal biopsy specimens were stained with hematoxylin-eosin, Giemsa and Azure II. Histologic changes included atrophy (3/14), duodenitis (2/14) or both (3/14). Transmission electron microscopy was used for the identification of developmental stages of Cryptosporidium sp. PMID- 9412139 TI - [Renal failure in patients with liver transplant: incidence and predisposing factors]. AB - Renal failure is a common finding in patients undergoing orthotopic liver transplantation. The aim of the present study was to evaluate the incidence, prognostic value of pre, intra and postoperative factors and severity of renal dysfunction in patients who undergo liver transplantation. Therefore, the records of 38 consecutive adult patients were reviewed. Renal failure was defined arbitrarily as an increase in creatinine (> 1.5 mg/dl) and/or blood urea (> 80 mg/dl). Three patients were excluded of the final analysis (1 acute liver failure and 2 with a survival lower than 72 hs.) Twenty one of the 35 patients has renal failure after orthotopic liver transplantation. Six of these episodes developed early, having occurred within the first 6 days. Late renal impairment occurred in 15 patients within the hospitalization (40 +/- 10 days) (Mean +/- SD). In he overall series, liver function, evaluated by Child-Pugh classification, a higher blood-related requirements and cyclosporine levels were observed more in those who experienced renal failure than those who did not (p < 0.05). Early renal failure was related with preoperative (liver function) and intraoperative (blood requirements) factors and several causes (nephrotoxic drugs and graft failure) other than cyclosporine were present in patients who developed late renal impairment. No mortality. No mortality was associated with renal failure. We conclude that renal failure a) is a common finding after liver transplantation, b) the pathogenesis of this complication is multifactorial and, c) in not related with a poor outcome. PMID- 9412140 TI - [Use of spasmolytic agent otilonium bromide (spasmomen) in digestive endoscopy: a prospective study in 63 patients]. AB - Otilonium bromide is a calcium antagonist with a direct myolytic effect, that is indicated in spastic conditions and functional dyskinesias of the gastroenteric apparatus (irritable bowel syndrome) and as a premedication for gastrointestinal endoscopic procedures. The present study assessed otilonium bromide 40 mg PO the night before and 40 mg PO the morning in 49 upper and 14 lower flexible endoscopies in 63 patients, to determine the presence or absence of peristalsis and relaxation of the pylorus. No side effects were observed due to the medication. In 46 (93.8%) upper endoscopies marked relaxation of the gastrointestinal tract and also pylorus relaxation were observed. In 13 (92.8%) lower endoscopies, marked relaxation of the colonic tract was also seen. All patients tolerated well the endoscopies. Otilonium bromide was useful as premedication in order to enable upper and lower endoscopic explorations, because of its spasmolytic effect. PMID- 9412141 TI - [Granulomatous colitis associated with diverticular disease. Report of a case]. AB - We report the case of a 71 years old lady with clinical and radiological evidence of colonic obstruction due to diverticulosis. A segmentary resection of the sigmoid colon was performed and the diagnosis of diverticulosis and Crohn's disease was made in the surgical specimen. The patient is doing well and no further extension of the colonic granulomatosis has been found 8 months after surgery. In spite of the great frequency of diverticulosis in the elderly its association with Crohn's disease is quite uncommon. This is probably due to the infrequency of colonic Crohn's disease in this population, particularly of the segmentary type. A few cases of segmentary colonic Crohn's disease that were cured after resection have been published. Even though the postoperative period of our patient is too short for making a good prognosis. Crohn's disease is a chronic relapsing disease and frequency of recurrences is closely related with long term evolution. PMID- 9412142 TI - [Esophageal squamous papilloma: a case report and review of the literature]. AB - We report a case of an esophagic squamous papilloma (SP) in a 90 year old patient. This pathology is benign and its diagnosis is generally a finding during an upper digestive endoscopy. Only 150 cases have been reported in the world's medical literature. Treatment consists in an endoscopic resection with diathermic snare or a conservative approach with periodical endoscopic control and observation depending of the type of patient. PMID- 9412143 TI - [Clinico-endoscopic spectrum of gastritis associated with Helicobacter pylori in pediatrics]. AB - In order to gain further knowledge about the clinical and endoscopic features of chronic gastritis (CG) associated with Helicobacter pylori (H py) we retrospectively analyzed 81 pediatric cases. All were biopsy-proven. The cases were divided in two groups: Group (1988-1992) included 21 cases. These represented the early stage of clinical recognition of the disease. Group 2(1993 1995) comprised 60 cases and represents the stage in which the disease was mandatory. Mean number of cases/year was 4.2 and 20 for group 1 and 2, respectively. Recurrent abdominal pain (RAP) was the most frequent clinical symptom (74/81; 91%), followed by upper digestive tract hemorrhage (UDTG) (34/81; 41.9%). The combination gastroesophageal reflux (GER) and esophagitis (E) was found in 52/81 (64.2%) of the children. Endoscopically, granularity of the mucosa was more frequently found in cases with RAP (47/74), GER (28/36) and E, while a smooth mucosa predominated in patients with UDTH (23/34). Our findings strongly suggest that symptomatic CG with H py in children expresses peculiar clinical and endoscopic features. Since RAP was present in 91% of the cases it appears adequate to include this disease in the differential diagnosis of it. These clinical manifestations have not been previously linked to CG with H py. Better understanding of the clinical and endoscopic spectrum of CG with H py results in adequate treatments and possibly prevention of gastric (and esophageal) diseases found in adults. PMID- 9412144 TI - [Gastroesophageal reflux disease refractory to medical treatment. Which approach: the pill or the scalpel?]. AB - We analyze the modern literature regarding natural history, clinical picture and treatment of gastroesophageal reflux disease. The characteristics of a small group of patients with tendency to relapse are emphasized, analyzing the possibilities offered by pharmacological as well as surgical treatment. Finally, we analyse the current literature of our country situation. PMID- 9412145 TI - [Helicobacter pylori in the pediatric population]. PMID- 9412146 TI - [Cryptosporidiosis in AIDS. A problem for clinicians and gastroenterologists]. PMID- 9412147 TI - [Duchenne type progressive muscular dystrophia]. PMID- 9412148 TI - [SPECT and PET in neurological practice]. PMID- 9412149 TI - [Results and considerations regarding auditory neonatal screening based on the use of transient evoked otoacoustic emission]. AB - The authors present a 5-year clinical experience in audiological screening performed at the neonatal center of the Policlinico of Perugia, Italy. The study was performed using an IL088 Otodynamics unit produced by Bray & Kemp. A total of 1328 newborns (2656 ears) were tested on the 4th day of life and during spontaneous sleep. None of the children had any audiological risk factors. The test was repeated one month later for all subjects who lacked Transient Evoked Otoacoustic Emissions and in many cases ABR testing was performed by 3 months of age. The authors present the undoubted advantages of the present method which include the fact that it is a) easy to perform, b) non invasive, c) sensitive and d) effective. They then discuss the main problems which arose during the course of the screening and advance some solutions. For the most part these problems involved the high number of false positives (13.1%) and the high percentage of subjects who were lost to subsequent controls (approximately 6% of the total population). The number of false positives could be reduced by using a linear acoustic stimulation (rather than the non-linear stimulation which is the default parameter for the machine). Such a linear stimulation can improve the signal-to noise ratio, thus making it possible to adopt a reproducibility index lower than the 70% presently used (however, this brings with it the risk of including a certain number of false negatives). Finally, they discuss the possibility of only retesting those subjects with bilateral lack of TEOAEs, thus reducing the number of check-ups to be performed a month later. PMID- 9412150 TI - [Annular wedge tympanoplasty: a variation of overlay myringoplasty]. AB - Myringoplasty has been increasingly refined in recent years and today the most frequently employed are the "overlay", the "underlay" and the "interlay". Of these the overlay technique appears to best guarantee graft stability. However, with this technique there is the risk of blunting and neotympanum lateralization which can compromise functional recovery. To obviate these drawbacks, the authors propose a modification of the classical overlay technique. This modification consists of detachment of the anterior portion of the Gerlach annulus and the adjacent protympanum mucosa in order to insert the graft between the bony and fibrous portions of the annulus. This technique is defined as the "Annular Wedge Tympanoplasty" (AWT). From January 1993 to July 1994 a total of 74 tympanoplasties were performed using the AWT technique to reconstruct the tympanic membrane. In 71 (96%) of these, the opening closed completely. As regards incomplete healing, 2 cases showed signs of blunting, 3 showed posterior lateralization and 1 full lateralization with a reduction in the hearing level recovery. The work is not conclusive although it does present a technique which is easy to perform and which provides good functional recovery. PMID- 9412151 TI - [Tryptase determination in nasal washings in patients with allergic rhinitis]. AB - Triptase is an enzymatic protein found in the mastocytes. Although the biological role of this substance is not fully understood, significant amounts of it are found in the nasal secretions of allergic subject both when naturally exposed to the allergen and when subject to specific nasal provocation tests. The present study uses a radioimmunological method (Pharmacia, Uppsala, Sweden) to assay the amount of triptase in two groups: one composed of 10 subjects suffering from allergic rhinitis and the other of 10 non allergic controls. All subjects were submitted to nasal wash with a saline solution prior to endonasal administration of an allergen extract (Poa Pratensis or Dermatophagoides Farinae). Fifteen minutes after nasal provocation the nose was once more washed out with a saline solution. All the wash liquids, both before and after stimulation, were assayed to determine the triptase content and the results for the two groups were compared. The results did not follow a normal distribution and were compared. The results did not follow a normal distribution and were processed with non parametric statistical methods. The allergic subjects showed base values, of 1.4 U/l (mean) while after allergic stimulation this jumped to 14.5 U/l (mean) (p < 0.005). On the other hand, the pre- and post-stimulation values were comparable for the non allergic group. In addition, there was no statistically significant difference between the basic pre-trial triptase levels in the allergic and non allergic subjects. The results indicate that assaying the triptase content in nasal wash liquid can provide a useful support in the diagnosis of allergic rhinitis. PMID- 9412152 TI - [Microsurgical treatment of ethmoid-sphenoidal fluid fistula]. AB - Traumatic basal skull fracture and iatrogenic injury during surgery are the main causes of cerebrospinal fluid (CSF) fistulas. The roof of the ethmoid and the cribriform plate are the most frequent sites of CSF rhinorrhea. The technique for repairing CSF leaks has evolved from intracranial repair to extracranial approaches. From March 1995 to June 1996 five patients with CSF rhinorrhea underwent microsurgical transnasal repair. In four cases of ethmoid defects, a pedicled vascularized mucoperiostal flap was obtained from the ipsilateral septum and placed over the defect. The fifth CSF leak came from the sphenoid and was repaired by packing the sinus cavity with abdominal fat. In all cases the CSF pressure was reduced with a lumbar drain for 5-10 days. The nasal packing was removed on the 5th day. All five patients have been followed up regularly for at least 6 months. To date there has been no evidence of recurrence. The surgical microscopic approach has some advantages: it permits good control of the surgical field and bleeding. In addition, stereoscopic vision provides the surgeon with a meticulous apposition between the flap and the CSF leak. The surgical technique is discussed in detail. PMID- 9412153 TI - [Quality of life after treatment in patients with laryngeal carcinoma]. AB - Today laryngeal cancer can be cured by means of a variety of treatments (nearly 60% of the patients in an unselected population are still alive after 5 years). Despite the low incidence, this form of cancer can present a significant social problem because the form of treatment can have an impact on the esthetic, functional and emotional aspects affecting the quality of life (QOL). In the present study 690 laryngeal cancer patients treated with 6 different forms of therapy (total laryngectomy, partial laryngectomy, cordectomy, radiotherapy alone, total laryngectomy plus post-operative radiotherapy, partial laryngectomy plus post-operative radiotherapy) were asked to fill out a specific EORTC CORE QOL Questionnaire (EORTC QLQ C-30) as well as a specific head and neck questionnaire. Six multi-item function scales, 3 symptom scales and 6 individual items assessing both symptoms and economic consequences of the disease were evaluated. A total of 517 patients (74.92%) filled out the questionnaire. For each form of therapy the patients were divided by age (under and over 65 years of age). The results indicate that the quality of life is better in those patients who underwent a single form of treatment (i.e. radiotherapy alone, partial laryngectomy, total laryngectomy) than in those who underwent combination treatments (i.e. surgery plus radiotherapy). Moreover, the results were better in the older patients. Quite often laryngeal cancer patients are subject to psychosocial problems although this did not show up in the present study where the patients tended to consider surgery as a liberation. The social-cultural level of the patient has a significant effect on the quality of life as it proved better in those social classes were physical strength is of prime importance as opposed to those dominated by social parameters such as socialization, communication and aesthetics. PMID- 9412154 TI - [Inhalation of foreign bodies: epidemiological data and clinical considerations in the light of a statistical review of 92 cases]. AB - In the present work 92 patients were studied all of whom had inhaled a foreign body (FB) into one of the tracheobronchial branch. The following factors were evaluated: sex, age, nature of the FB, localization in the respiratory tree, clinical symptoms, radiological findings, time lapse between diagnosis and removal. The peak incidence (61.9%) was in children under 3 years of age with a male-female ratio of 2:1. The most frequently inhaled FBs were of organic nature (31.5%); of these 58.6% were peanuts. The time lapse between inhalation and removal of the FB was as follows: in 20.5% the object was removed within 24 hours; in 66.4% within one week; in 12% in more than a week; and in 1.1% it took more than 8 weeks. In 53.2% of the cases the right bronchial branches were involved while in 28.2% the left side was affected. The most frequent symptoms were coughing (73.9%), wheezing (69.5%), dyspnea (51%) and fever (17.3%). Radiography detected the FB in only 7 cases (8.7%); in the remaining cases only indirect signs of the FB could be found: atelectasia (11.9%), emphysema (19.5%), cardio-mediastinic shift controlateral to the FB (10.8%). As regards complications, only 6 patients showed signs of slight endobronchial bleeding, 2 cases showed a pneumothorax and one other patient required a tracheotomy because of the particular shape of the FB which proved unable to pass backward through the glottis. In all cases the FB was removed using stiff bronchoscopy under either local or general anesthesia. The authors feel that, even if no clinical signs are found and radiography proves negative, one must always consider the possibility of a FB in the tracheal-bronchial branches, particularly in patients within the age range most at risk (under 3 years) and in those having a highly suspicious clinical history. In addition, the authors assert that the use of corticosteroids before and after the bronchoscopy markedly decreases the incidence of post-operative subglottic edema which would require an emergency tracheotomy. PMID- 9412155 TI - [Doppler-color ultrasonography in the diagnosis of parotid tumors]. AB - Today ultrasound, often integrated with CT and NMR, is used in the differential diagnosis of parotid masses. Color-Doppler Sonography is a recently introduced method which makes it possible to evaluate intra- and perilesional vascularization and to perform a hemodynamic study of the area being explored. Using an Acuson 12P ultrasound unit 21 patients with surgically treated parotid neoformations were examined. All had previously undergone traditional ultrasound and CT. An abundant vascularization, both intralesional and peripheral, was found in 4 malignant lesions, in one intraglandular granulomatose lymph adenopathy, in one pleomorphic adenoma of the deep lobe and in one case of Sjogren's syndrome. In the remaining cases (7 pleomorphic adenomas and 7 cystoadenolymphomas) vascularization was either totally absent or only slightly visible in the periphery. These preliminary results would appear to indicate that the presence of hypervascularization testifies to a malignant lesion or to an inflammatory disorder. The Color-Doppler Sonography can provide a useful adjunct to conventional ultrasound, increasing diagnostic accuracy in cervical-parotid masses. PMID- 9412156 TI - [Parotid gland sialolithiasis: a new therapeutic option]. AB - In chronic lithiasis of the salivary glands surgery is the only approach available whenever the stone cannot be removed by dilating or dissecting the salivary duct. The introduction of lithotripsy in the treatment of kidney and gall stones has made it possible to apply this method to salivary gland stones as well. The present study was aimed at evaluating the safety, reliability and effectiveness of lithotripsy in 36 patients with symptomatic parotid gland stones which could not be removed by conservative surgery. Ultrasound revealed the stones which were on the average 4.8 mm. In 147 cases they were located in the intraparenchymal area while the remaining 19 cases were located within the duct. No patient required anesthesia, sedative or analgesic. The average number of sessions per patient was 3.8 and each treatment lasted an average of 30 minutes. After lithotripsy, in all 10 of the patients who had completed the treatment, the stone was either totally eliminated or reduced enough to allow for spontaneous elimination. The side effects found in 9 of the patients were minor pain (2 patients), a transitory duct hemorrhage (6 patients) and skin petechia (4 patients). No damage to the salivary glands or adjacent anatomical structures was revealed upon treatment or later during the 10 month average follow up (range 1 22 months). Extracorporeal lithotripsy is a safe, effective, non-invasive alternative therapeutic option for the treatment of parotid sialolithiasis. PMID- 9412157 TI - [Benign symmetrical lipomatosis: Madelung's disease]. AB - Multiple symmetrical lipomatosis, or Madelung's disease, is a rare disease of unknown etiology. It is characterized by the presence of loose adipose tissue deposits localized in the cervical region and in the upper body. The neoformations grow slowly and their initial consequence is purely esthetic. They can, however, lead to compression of the laryngeal-tracheal area and of the esophagus. This disease normally affects middle-aged males from the Mediterranean area with a history of alcohol abuse. Although most cases have been sporadic, a few authors have indicated that the disorder may be hereditary. It is thought that this pathology originates from an alteration in lipid metabolism. Surgical removal of the lipomatose mass is the treatment of choice even though there are frequently recurrences. A case is presented of a rare laryngeal localization of this disease and diagnosis and treatment are discussed. PMID- 9412158 TI - [Influence of hypoxia on the human auditory system]. AB - The present paper presents a review of the literature on "hypoxia and human auditory mechanisms". It examines and discusses, above all, the results obtained in the various studies using pure tone audiometry and auditory evoked potentials. At the present time, the two areas which appear most sensitive to hypoxia are the cochlea and, above all, the telencephalic auditory cortex (specifically those sectors dedicated to cognitive processing of auditory stimulation). However, many other areas which are sensitive to hypoxia, but to a lesser extent, have also been identified, even in other sectors of the auditory pathway. Particularly worthy of note is the effectiveness of the metabolic compensatory mechanisms which come into play upon hypoxic stress. These mechanisms include vasodilation and the presence of metabolic reservoirs. Nevertheless, there are still a number of open questions regarding how the auditory pathway functions in the case of hypoxia; thus the experimental study of hypoxic hypoxia is still an interesting, fruitful research field in audiology. PMID- 9412159 TI - [Psychobiological approach to personality disorders]. AB - Current categorical classifications of the personality disorders are based on psychological perspectives. Consequently, biological research using these models has been unsuccessful since the nosological categories do not come out from the psychobiological systems underlying personality psychopathology. By contrast, biological research on dimensional models of personality has been more useful. All dimensional models agree on a neurotic-inhibitory dimension and an exploratory dimension of personality but they differ on other primary traits like dependence, emotionality or impulsiveness. However, gathering together all biological data support the existence of five biological axes of personality: a cognitive axis, a mood axis, an anxious-inhibitory axis, an exploratory axis and a action-control axis. CONCLUSION: Considering these biological axes is bringing a new perspective on the classification of personality disorder and gives way to new pharmacological therapeutic options. PMID- 9412160 TI - [Predictors of rehospitalization in schizophrenia]. AB - OBJECTIVES: The aim of this study is to determine the predictive value on rehospitalization of sociodemographic variables, positive/negative symptoms and thought disorders. The results are part of research project founded by the Basque Health Department. METHODS: A 18 month follow-up study of a cohort of 60 patients with acute exacerbation of schizophrenia was carried out. The assessment was performed with DSM III-R diagnostic criteria, PANSS and CGI rating scales, and SCID-P semistructured interview. All patients received antipsychotic treatment. The sociodemographic and disease data, the dimensional score of the PANSS subscales, the score of CGI scale, the items 2, 12, 13 and 14 of the PANSS as indicators of formal thought disorders; and the items 1, 5, 6, 17 and 23 of the PANSS as content thought disorders were established as predictors. The predictive value was determined by the Cox regression test (Lee 1992). RESULTS: We did not find predictive value either in the PANSS scores or in the 9 thought disorders evaluated (Wald and RR tests were not significative). Nevertheless, considering the values of standard error obtained in the Cox regression we were not in a position to assure that they did not have an incidence in the hospitalizations. The CGI was the only scale that showed prognostic value (Wald test = 1.9945; RR = 1.7499). Our results indicated that the lower number of previous hospitalizations (Wald test = 1.1437; RR = 1.1437) and the high level of studies (Wald test = 2.4258; RR = 1.8052) diminished the risk of rehospitalization. CONCLUSIONS: 1 o The predictive value on rehospitalization for the positive/negative symptoms and thought disorders was not confirmed. 2 o CGI is the only scale with predictive value. That fact makes us consider the importance of what German psychiatrists called "smelling the schizophrenia" or "The smell of schizophrenia". 3 o Our results indicate that the lower number of previous hospitalizations, and the high level of studies diminish the risk of rehospitalization. PMID- 9412161 TI - [Cost effectiveness analysis of olanzapine versus haloperidol in the treatment of schizophrenia++ in Spain]. AB - BACKGROUND: Cost-effectiveness of olanzapine in comparison with haloperidol, in Spanish schizophrenic patients, was analysed using a clinical decision model. METHODS: The model represents a simulation of the different clinical and therapeutic possibilities that an hypothetical cohort of patients could experienced in a 5-years period of treatment. Efficacy was measured as months with partial-complete remission. Most information was obtained from the HGAJ randomised clinical trial. Other information was estimated through literature reviews and the opinion of an expert panel. RESULTS: Average cost-effectiveness for olanzapine was lower (116,476 pesetas per month with partial-complete remission) than for haloperidol (134,762 pesetas per month with partial-complete remission). Olanzapine produced more than half year (6.7 months) with partial complete remission, in comparison with haloperidol, with antincrementar cost effectiveness of 32,516 pesetas per month with partial-complete remission, in comparison with haloperidol. The results were not sensitive to changes in the values of the main variables used in the analysis. CONCLUSIONS: According to this analysis, olanzapine presents a good cost-effectiveness relationship in comparison with baloperidol, in Spanish schizophrenic patients. The analysis will be completed when new studies comparing olanzapine with other antipsychotics are available. PMID- 9412162 TI - [Premenstrual dysphoric disorder: long-term treatment with fluoxetine and discontinuation]. AB - INTRODUCTION: Premenstrual Dysphoric Disorder (PMDD) is characterized by debilitating mood and behavioral changes in the weed preceding menstruation that interfere with normal functioning. The diagnosis requires that symptoms be recurrent and persistent. We report here the results of an open-label study with 20 patients with PMDD according DSM-IV with and without a concomitant diagnosis of MDD (Major Depression) treated with fluoxetine during 6 and 18 months. METHOD: 9 patients with PMDD and MDD, and 11 patients only with PMDD were treated with fluoxetine 20 mg/d during 6 months. After that, 9 women continued treatment until complete 18 months of treatment and 11 women discontinued by their own decision, because the complete absence of symptomatology. Women who discontinued treatment were followed at the 18 months to the assessment of the evolution. The efficacy was assessed by CGI, HAMA, HAMD and un TDP scale monthly during the first 6 month and in the visit at 18 month. RESULTS: All patients completed the 6 months of treatment with a good tolerance. The most common adverse events were decrease of the libido, gastric complains, nervousness and insomnia. These events were mild and don't produce any withdrawn. There were a significant change in all scales at 6 month assessment: HAM-D (22.4 vs. 5.3, p < 0.000), HAM-A (23.8 vs 8.7, p < 0.000) and TDP (36.4 vs. 18, p < 0.000). There weren't differences among patients with and without concomitant diagnosis of MDD. Among patients who continued taking fluoxetine the improvement was maintained at the 18 month-assessment. Patients who discontinued treatment with fluoxetine after 6 months suffered a worsening in their symptomatology at the 18 month assessment (CGI = 3.18 vs. CGI = 4.81, p < 0.005). CONCLUSIONS: Fluoxetine is an effective and well-tolerated treatment for the long treatment of PMDD. The remission is maintained at least during 18 month of treatment. Symptomatology reappears after treatment discontinuation in most of the women. PMID- 9412163 TI - [Validation study of the Center for Epidemiological Studies-Depression of a Spanish population of patients with affective disorders]. AB - The Center for Epidemiologic Studies-Depression (CES-D) is a short self-rating scale composed of 20 items, designed to detect depressive symptomatology. It has demonstrated its sensibility in psychiatric patients and general population. The CES-D was administered to 99 patients, 33 men and 66 women with mean age of 44.14 years. The patients had been diagnosed of: Major Depressive Disorder (74%), Bipolar Disorder (10%), Adaptive Disorder with depressive mood (10%) and other mood disorders (6%) according to DSM-IIIR criteria. In order to study the validity and reliability of the CES-D, we administered the Hamilton Rating Depression Scale, the Beck Depression Inventory and analogical scales to all the patients. In the reliability analysis we obtain 0.9 alpha. The factor analysis show 4 factors who explain the 58.8% of the variance: "depresses Affect/Somatic", "Positive Affect", "Irritability/Hopelessness", "Interpersonal/Social". The scale shows a 0.95 sensibility and 0.91 specificity to depressive symptomatology detection (according to scores equal or over 9 on HRSD) taking as cutoff scores equal or over 16 on CES-D. Our results show that the CES-D is a sensitive and specify tool for depressive symptomatology detection in psychiatric population. The CES-D is easy to be completed and evaluated, therefore may be useful in epidemiologic studies in general populations. PMID- 9412164 TI - [Critical evaluation of the use of antidepressants in adolescence]. AB - In the introduction the authors highlight the clinical theoretical and practical importance of affection disorders, especially depressive ones, during childhood and adolescence, their relationships of continuity and the limitations of psychopharmacological studies with antidepressants and other medicines. The authors review the bibliography consulted about the use of antidepressants in depressive disorders during adolescence following this order: authors and years, medicine used, type of population, number of cases, proportion of males and females, age ranges, dose in mg/kg/day, ranges of plasmatic concentrations, most frequent secondary and unwanted side effects, time of duration of the study, comorbidity, existence of a previous washing stage, as well as placebo and global results. The studies with triciclical antidepressants (TAD) are classified into open ones and controlled ones. Among the open ones, they review the studies by Dugas et al (1980), Geller et al (1985), Ryan et al (1986), Strober et al (1990), and Ambrosini et al (1994). Among the controlled ones, they review the studies by Kramer and Feiguine (1981), Geller et al (1990), Boulos et al (1991), and Kutcher et al (1994). In the third part, the studies of enhancement of TAD with lithium by Ryan et al (1988 a and b) and Strober et al (1992) are analysed. In the fourth part, the studies of enhancement of TAD with the IMAO by Ryan et al (1988-1990) are evaluated. In the fifth part, the ten studies with ISRS (fluvoxamine, fluoxetine, paroxetine and sertraline) on the treatment of depression during adolescence are also discussed. In the final comments there is a summary of the clinical perspective of this kind of psychopharmacological therapy. Four tables and 73 bibliographical references are included. PMID- 9412165 TI - [Mental disorders due to lacunar infarcts. Two case reports]. AB - Lacunar infarcts have a high prevalence, about 10%, mainly in the elderly. Although the most frequent feature are the lacunar syndromes as a neurologic form, psychiatrist should bear them in mind because it is possible the presence of psychiatric symptoms as the only manifestation of them. We report two examples of it. The first case in a man with a transitory episode of uninhibition, with maniform characteristics, and frontal disturbance signs. The second one is a woman with a peduncular ballucinosis. The only neuroradiological finding were lacunar infarcts in both of them. We discuss a pathogenic hypothesis to explain these features. The first case may be a disconnection of the system that integrates the basal ganglia, thalamus and thalamocortical projections; the second one can be originated because of interruptions of the nigroestriatal connections. PMID- 9412167 TI - [Pharmacologic treatment of benign prostatic hyperplasia]. PMID- 9412166 TI - [The psychiatric patient with deficits: a study of patients presenting at the emergency medical service]. AB - We present the six months prospective study results assessing clinical, social and demographic features in defectual psychiatric patients (chronic psychosis, dementia syndrome, intellectual deficit) who went to Basurto Hospital's Emergency room during that period of time. The studio data were obtained applying a clinical epidemiological questionnaire. Some problems derived from attending these patients were analyzed. Suggestions to improve the efficacy of the defectual psychiatric patient's assessment are proposed. PMID- 9412168 TI - [Surgical and urologic knowledge in Spain in the Renaissance: a vision from Santiago de Chile]. PMID- 9412169 TI - [Effects of vasectomy on the testicular structure of the dog]. AB - Study of the effect on testis structure in the dog caused by a vasectomy. Emphasis is made on how in animals undergoing ligature of both ends of the ductus deferens, existence of structural changes, which are consistently present, are proportional to the time elapsed from performance of vasectomy. Such changes are revealed by: degeneration of germinal epithelium, thickening of basal membrane and hypertrophy of interstitial tissue at the expense of unspecific connective tissue. Changes were first seen as from the fourth month, but a full year has to elapse prior to noting a marked germinal hypoplasia. These experimental results cast doubts on the safety of such sterilization approach, although they cannot be entirely extrapolated to humans. PMID- 9412170 TI - [Urinary excretion of cytokines in bladder carcinoma]. AB - OBJECTIVE: Analysis of the urinary excretion of cytokines in vesical carcinoma. MATERIAL AND METHOD: The study includes the results obtained in the quantification of several interleukins (IL-1, IL-2, IL-4, IL-6, INF-gamma and TNF alpha) in urine samples corresponding to 60 patients with transitional cell carcinoma (TCC) with vesical location (40 surface and 20 infiltrant). Concurrently, 40 healthy donors and 20 patients with urinary tract infections were studied. Determination of the various cytokines was done with ELISA enzyme linked immunoassays. RESULTS: The results obtained in the study show that: a) urinary concentrations of IL-1, IL-2, IL-6, TNF- and INF- in surface TCC, are similar to those found in healthy subjects; b) levels of the mentioned cytokines are significantly higher in patients with urinary infections; c) in patients with infiltrant TCC, IL-6 urinary concentration is significantly higher than in those with S-TCC; d) urinary IL-4 levels show no difference between the various groups under study. CONCLUSION: From all the above it is concluded that there is a large diversity in the excretion of urinary cytokines from the vesical urothelium based on antigenic stimulation (bacterial or tumoral) to which it has been exposed and the tumoral stage, and that baseline determination of IL-6 urine level in patients with vesical TCC could have some prognostic influence. PMID- 9412171 TI - [Uro-oncologic registry in the area of El Bierzo. 1990-94]. PMID- 9412172 TI - [Bladder neoplasms in patients under 40 years of age]. AB - The incidence, presentation and clinical evolution of vesical tumours diagnosed in our Centre over the last 10 years (1986-1995) in patients under forty are examined. This retrospective study included 19 of the 643 neoplasias treated during that period; 7 of these in patients under 30 and 12 in patients over 31. Haematuria was the most frequent reason for visiting the Centre. At the time of diagnosis, all cases were surface tumours and none progressed to the infiltrant stage. Tumours in patients over 30 are characterised by a higher histological grade and greater relapse rate versus those in patients under 30. Therefore, age may be a favourable prognostic factor in patients under 30 versus the older group who follow a course more similar to the remainder of usual patients. Diagnosis and treatment of these neoplasias should be just the same as for tumours in older patients. PMID- 9412173 TI - [Ultrasonographic and pathologic differences between non-palpable prostate cancer (T1c) and palpable lesions detected with rectal examination]. AB - OBJECTIVE: To determine whether non-palpable cancers (T1c) have different ultrasound and pathological features from other palpable cancers of the prostate gland. MATERIAL AND METHOD: PSA levels, ultrasound features and Gleason score in 178 patients diagnosed with prostate cancer between 1994-1995 were compared. Correlation of pathological findings in surgical sections from 47 patients undergoing radical prostatectomy. RESULTS: 22% tumours were non-palpable. No difference was observed between both age groups (p = 0.5) and PSA levels (p = 0.09). Differences were noted in favour of palpable cancer in PSAD (p = 0.01), incidence of ultrasound nodes (p < 0.001), capsule changes (p < 0.001), seminal vesicles (p < 0.001), Gleason score (p = 0.006) and bone scan (p < 0.05). Two (14%) patients with non-palpable cancer showed no tumour in the prostatectomy section. Apart from these 2 patients, no differences were found between both groups in terms of Gleason score (p = 0.3), local stage (p = 0.7) and node involvement (p = 0.4) in patients undergoing radical prostatectomy. CONCLUSIONS: 86% patients with non-palpable prostate cancer has clinically significant tumours and showed no differences from the rest of tumours undergoing radical prostatectomy. PMID- 9412174 TI - [Iatrogenic splenectomy in surgery of renal tumors]. AB - A description of our experience with iatrogenic spleen lesions requiring splenectomy caused during surgery for a renal tumour. Out of a total 83 nephrectomies due to renal tumour performed in our Centre between 1988 and 1996, 46 were left and 4 of them (8.7%) involve splenectomy. Clinical, surgical, pathoanatomical and follow-up data of these 4 cases is analyzed. Anatomical relationship of the spleen with kidney and colon, as well as factors to consider during surgery to avoid splenic lesions are discussed. PMID- 9412175 TI - [Extracorporeal shock-wave lithotripsy in the treatment of calculi in horseshoe kidneys. Our experience]. AB - Thirty-one (31 renal units (RU) were treated with extracorporeal shock wave lithotrity (ESWL) in 27 patients with lithiasis in horseshoe kidneys (HK). Treatment was done in an out-patients environment and in two thirds of cases the patient was placed in prone-decubitus. An average of 2.42 sessions per patient (range 1-8) were performed freeing 15 RU, 10 with removable residues, and failure in 6:3 with non-removable residues in all asymptomatic cases, 2 where fragmentation was insufficient and 1 renal annulment post-ESWL. Overall, results were "good" in 80.6% cases, with a significant relationship between size of lithiasis and therapeutical results. Thus, for < or = 2 cm calculi the result was good in 100% cases but a greater number of cleaning were required in those < 1 cm (62.5 vs 46.15%). On the contrary, "therapeutic failures" were limited to just > 2 cm calculi. It can therefore be concluded that most patients with lithiasis in HK, specifically when calculi are < 2 cm, can be treated primarily and in monotherapy with ESWL. PMID- 9412176 TI - [Leydig cell tumor associated with epididymal cyst]. AB - Case report of a Leydig's cell testicular tumour associated to an epididymal cyst in a young adult. The literature revealed no relationship between both conditions. Review of diagnostic and therapeutical approaches. PMID- 9412177 TI - [Acute scrotal hematoma in Schoenlein-Henoch purpura. Infrequent urologic manifestation]. AB - Schonlein-Henoch purple is a systemic vasculitis due to hypersensitivity. It is typical of childhood and more unusual in adults. The etiology is unknown. It is identified by the presence of skin purpura, joint involvement, abdominal colic pain and quite often renal involvement (nephropathy). Urological manifestations are uncommon and include: haematuria, scrotal swelling, cord haematoma, signs and symptoms mimicking cord or hydatid torsion, testicular pain due to intratesticular segmented infarction, painful ecchymotic induration of the scrotum, or testicular necrosis. Contribution of one case of Schonlein-Henoch purpura developed in adulthood with scrotal-perineal haematoma associated to skin purpura and abdominal colic pain in a 75-year old patient. PMID- 9412178 TI - [Small cell carcinoma of the bladder. Report of a new case]. AB - Contribution of a case report of vesical small cell carcinoma (SCC) seen at our Centre and managed with partial cystectomy and systemic chemotherapy (CMT) with M VAC. SCC is an uncommon neoplasia of the bladder usually associated with an aggressive behaviour. The effectiveness of radical surgery has not been demonstrated, so a conservative treatment was chosen which has allowed to preserve a high quality of life until now, 36 months after diagnosis. The studies of local and distant relapses show no signs of residual disease. PMID- 9412179 TI - [New report of epididymal adenomatoid tumor. Infrequent pathology]. AB - Case report of a 42-year old patient with an epididymal adenomatoid tumour found after a three-month history of increased left hemiscrotum and scrotal discomfort. Definite diagnosis was arrived at following surgical exeresis and pathoanatomical study. The etiopatogenesis, clinical presentation and management are all analyzed. Since urological incidence is very low within intrascrotal processes, we believe it is important to understand the condition so that a differential diagnosis from other inflammatory processes can be established; also the convenience of a much more aggressive attitude in scrotal diseases is restated due to the low morbidity of this type of surgery. PMID- 9412180 TI - [Bladder hemangioma: a clinical case]. AB - Contribution of a new case of vesical haemangioma in a 65-year old male. This is a benign and very rare tumour with not even 90 cases described in the urology literature to date. Haematuria was reported in all cases. This paper presents a review of the literature and a discussion of clinical and pathological features as well as diagnostic methods and recent changes in management approach. PMID- 9412181 TI - [Vesicovaginal fistula caused by foreign body in the vagina]. AB - Case report of a vesicovaginal fistula caused by the long-standing presence of a foreign body in the vagina. A routine examination verified the subsequent incontinence but it was only under general anaesthesia when a completely calcified plastic stopper was found in the vaginal fundus. PMID- 9412183 TI - [Sertoli cell tumor of the testis: clinical pathologic varieties. Report of a case]. AB - Tumours derived from sex cords and primitive gonadal stroma account for 4% of total testicular tumours. The low frequency of Sertoli's cells tumour (SCT) and the uneven study and follow-up of patients makes analysis of this tumoral entity difficult. This paper contributes one case report of a Sertoli's giant cell tumour calcified in a 13-year old patient, and reviews the clinical aspects, clinico-pathological varieties believed to require assessment in patients with this type of disease. This type of tumour is considered benign in its biological behaviour, although some malignant forms have also been described. SCT is actually an heterogeneous tumoral pathogenic entity with regard to pathogenic and prognostic aspects. Our final conclusions show that the clinico-pathological variety, age, size and associated clinical manifestations appear to be related to the prognosis. PMID- 9412184 TI - [Primary retroperitoneal hydatidosis]. AB - Genitourinary involvement by the Echinococcus granulosus larvae (urinary hydatidosis) ranks third in order of frequency after liver and lung involvement. The finding of a primary hydatidic cyst with retroperitoneal location is an uncommon fact. This paper presents once case of this infrequent disease. A revision of the different etiopathogenic mechanisms, as well as diagnostic and therapeutic approaches is made. PMID- 9412182 TI - [Vesico-scrotal hernia. Report of a clinical case]. AB - Incidental discovery of vesical hernias during herniorrhaphy is quite common. In such cases, patients are usually asymptomatic since the hernia portion is small and easily repairable. On the other hand, vesical solid protrusion to the scrotum is quite unusual, and is generally found associated to obstructive urinary symptoms. Management involves basically the correction of any associated obstructive conditions, correction of the vesical hernia and herniorrhaphy. PMID- 9412185 TI - [Changes in the caudal regression of fused Muller ducts: utriculocele. Three clinical forms of presentation]. AB - Utriculocele is the engrossment of the prostate utricle with occlusion and/or cystic formation, sometimes including the ejaculatory conducts. We report three patients diagnosed with utriculocele with different presentation forms: hypospermia, haematuria and mictional syndrome, and the treatment used. Theories on their formation, clinical presentation forms described in the literature, diagnostic means, most frequent associations, and different therapeutic alternatives are all discussed as well as a revision of the reported cases. PMID- 9412186 TI - [Molecular staging in urologic tumors. From wish to reality]. PMID- 9412187 TI - [Antimicrobial prophylaxis in urology]. AB - Antimicrobial prophylaxis in surgery has proven to be effective in controlled randomized trials. Usage in Urology is known at least since the '30s although its effectiveness has only become known since 1979. METHODS: Review of literature related to surgical antibiotic prophylaxis, more specifically urological surgery, basically from 1991 to 1995, but without overlooking those papers that have become classics due to their impact. RESULTS AND CONCLUSIONS: Efficacy of antimicrobial prophylaxis in urological surgery is nowadays beyond all doubt. Usage is indicated in the presence of sterile urine and dosage must be short, in single dosis in the immediate pre-operative or within 24 hours after the procedure. However, there is a number of issues that deserve to be treated in more detail for better understanding. Those are the establishment of adequate prophylactic regimes in renal transplantation and the use of antimicrobials based on their pharmacokinetic characteristics to optimize the prophylactic purpose. PMID- 9412188 TI - [Experience with the of treatment renal cell carcinoma with thrombus in inferior vena cava and right atrium]. AB - Retrospective analysis of definite staging and surgery results in 17 patients with renal cell tumour disseminated to lower cava vein who underwent radical nephrectomy and tumoral thrombectomy. Magnetic resonance predicted presence and level of tumoral thrombus in 100% and 88% cases, respectively. Neither venacavography or doppler echography provided additional information. Dissemination was infrahepatic in 9 (53%) cases, suprahepatic in 4 (24%) and to the right atrium in 4 (23%). Cardiopulmonary by-pass and hypothermic cardioplegia was used in 9 (53%) cases. Operative mortality and morbidity rates were 11% and 65%, respectively. The level of the thrombus did not significantly affect the prognosis which was highly affected however by regional node invasion. In all, CT and MRI can determine the extension and level of the cava vein thrombus in most cases. In our experience, disease-free survival is determined by the pathological stage and not by the extent of the cava thrombus. Radical nephrectomy and tumoral thrombectomy can provide long survival to patients with locoregional disease. PMID- 9412189 TI - [Chromophobe renal cell carcinoma: immunohistochemical study of 7 cases]. AB - A series of 7 cases of chromophobe cell carcinoma with clinicopathological correlation is studied. The immunohistochemistry analysis was carried out evaluating in all cases: low and high cytoqueratines (AE1, AE3), membrane epithelial antigen (EMA), vimentin, carcinoembrionary antigen (CEA), proliferative cell nuclear antigen (PCNA) and the oncoproteins p53 and focal c erbB2. In all cases a diffuse positivity for EMA, AE3 and focal for AE1 was observed, while vimentin and CEA were negative. The positivity for PCNA was always less than 25% of tumoral cells. The oncoprotein p53 was in all cases negative and only in one case a mild and focal positivity for c-erbB2 was observed. CONCLUSIONS: 1) The negativity for vimentin and positivity for EMA, AE1 and AE3 allows to differentiate this carcinoma from clear cell carcinoma. This corroborate a distinct histogenesis, thus suggesting their origin in the intercalar cells of distal collector tubules. 2) The low values of PCNA and the negativity for both oncoproteins p53 and c-erbB2 supports the low aggresivity of this tumor and a better prognostic than other types of kidney carcinomas. PMID- 9412190 TI - [Yield of transrectal ultrasonography in the diagnosis of prostatic cancer in symptomatic patients with normal rectal digital test]. AB - The purpose of this retrospective study is to evaluate the incidence of prostate cancer in symptomatic patients with non-suspect rectal examination and its correlation to PSA, PSAD levels and ultrasound findings. A total of 235 patients with non-suspect rectal examination underwent transrectal ultrasound and echo guided prostate biopsy to assess an echographic node, PSA > 10 ng/mL and/or PSAD > 0.15 Incidence of prostate cancer was 16% and no correlation was seen to either PSA (95CI = 5%, 14.9%) or the existence of ecographic nodes (95CI = 5.2%, 22.2%), mainly in the subgroup of patients with PSA > 10 ng/mL and no identifiable echographic node (95CI = 5.5%, 29.5%). A PSA > 10 ng/mL or identifiable echographic nodes in symptomatic patients with non-suspect rectal examination did not involve a risk factor for prostate cancer, however a PSAD > 0.15 within the group with PSA > 10 ng/mL with no echographic nodes did involve a risk factor. PMID- 9412191 TI - [Partial cystectomy as treatment of infiltrating transitional carcinoma of the bladder]. AB - The most widespread opinion, and until recently the only option, is that every vesical transitional cancer invading the muscle is, regardless its extent, candidate for radical cystectomy and that in spite of nobody questioning the advantages of partial cystectomy. MATERIAL AND METHODS: 45 patients with vesical infiltrant cancer T2 or higher, followed between 9 and 258 months and managed with partial cystectomy, were analyzed. Only patients with no radiotherapy were included and only in one patient pre-operative chemotherapy was used. RESULTS: In 8 patients no tumour was found in the specimen (pTO). Tumour grade was pTa in 2; pT1 in 11; pT2 in 5; pT3a in 4; pT3b in 11; and pX in 4 patients. Eight (8) patients had nodal involvement. Twenty-one (21) cases showed bladder relapse. In six (6), vesical infiltrant relapse was associated to metastasis. One case showed vesical relapse, pelvic mass and metastasis, and 4 only metastasis. Extravesical disease-free time and survival are better than in the group treated with radical cystectomy. But this is a highly selected group. CONCLUSIONS: With the same prospects of extravesical disease-free time and survival we offer: shorter, less risky surgery with low post-surgical morbidity and mortality and less hospitalization and proportion of late sequela. Better quality of life, with no skin stoma, incontinence or impotence Although the risk of vesical relapse persists, the procedures required to resolve vesical shunt or replacement complications are more aggressive than TUR sufficient to treat most relapses, and when recurrence is infiltrant radical cystectomy may be used as a rescue measure. This is so even now with the profusion of the so-called "mini-invasive" procedures. We believe that neither radio- and/or chemotherapy combinations contribute nothing to partial cystectomy alone. They may even be harmful and have significant side-effects. It is plain that POs are the result of total removal by TUR. Due to the little reliability when defining T, it is very hard to evaluate the contribution of adjuvant measures. Patients with no vesical tumour (pTOs) or pT1-pT2 tumours, and even up to pT3a, should not be included in protocols to evaluate the efficacy of combined cytostatic agents since their use is superfluous. Radiotherapy makes no contribution to this type of tumour in terms of local relapse and apparently has no effect on the metastasis. PMID- 9412192 TI - [Effectiveness of transrectal ultrasonography in local staging of prostatic cancer]. AB - OBJECT: To evaluate the value of rectal examination, transrectal ultrasound and their association in patients with prostate cancer undergoing radical prostatectomy. MATERIAL AND METHODS: Retrospective study on 56 patients who underwent radical prostatectomy between 1995-1995, mean age 63.2 +/- 10 years, to compare local clinical staging of extracapsular dissemination and seminal vesicle invasion performed by rectal examination and transrectal ultrasound with pathological findings in the prostatectomy specimens. The sensitivity, specificity, VPP, VPN, accuracy and percentage of under- and over-staged patients were assessed. RESULTS: Prevalence of locally advanced disease was 48%. Sensitivity to detect extracapsular dissemination was significantly higher with ultrasound (38%) than rectal examination (6%). Association of both techniques increased the sensitivity (50%) though not significantly; sensitivity to detect vesicle invasion was very low (14%). Accuracy of ultrasound to establish an overall definition of the local stage was 66%, but 59% patients with locally advanced disease were understaged and only 10% patients with localized disease were overstaged. CONCLUSIONS: Transrectal ultrasound showed low sensitivity to define the locally advanced disease mainly at the seminal vesicle level, with acceptable specificity. Overall evaluation of findings with RE and TRU increase discreetly the efficacy of prostate capsule staging. PMID- 9412193 TI - [Cervico-urethro-suspension by the Raz's technique in the treatment of stress urinary incontinence in women]. AB - Presentation of our results in the treatment of urinary exertional incontinence in women using Raz's cervicourethral suspension. From January 1991 through December 1995, 87 patients were operated (mean age: 55.64 years; range 36-74). Mean follow-up was 29.4 months. Recovery from incontinence or permanence of minimal occasional leaks due to major exertion were rated as good results and were achieved in 75 cases (86.20%). Percentage of success in patients with mild incontinence was 93.33%; 88.88% in moderate incontinence; and 58.33% in severe incontinence, differences being statistically significant (p < 0.01). No statistical significance was found relative to age, prior incontinence corrective surgery, hysterectomy or association with urgency incontinence. Prior to surgery, 21 patients also had a component of urgency incontinence which disappeared post surgery in 18 (85.71%) cases. De novo urgency incontinence appeared in 4 (6.06%) cases. Complications seen were 3 vesical perforations (3.44%). 1 urethrovaginal perforation (1.15%), 2 enterocele (3.44%) and 24 patients with transient urinary retention (27.58%). We believe this technique offers long-term successful results with a moderate morbidity rate. PMID- 9412194 TI - [Autoplastic closure of vesicovaginal fistulae with posterosuperior bladder flap (Gil-Vernet operation)]. AB - OBJECTIVE: To describe our experience in the treatment of vesico-vaginal fistula with autoplastic closure using a postero-superior vesical flap. Also, to discuss the technical details of the procedure. MATERIAL AND METHOD: Between 1985 and 1996, 15 patients with vesico-vaginal fistulae secondary to gynaecological surgery were operated. The fistula was considered complex in 5 patients based on previous attempts for surgical closure, number of openings or because they were located in the posterior gradient of the vesical neck. In all cases, autoplastic closure of the fistula was done with a vesical flap through an extraperitoneal abdominal approach. RESULTS: In 100% cases closure of the fistula was achieved at the first surgical attempt. There was no need to interpose vascular tissue between the bladder and the vagina to secure closure of fistula. Post operatively, 5 patients showed decreased vesical capacity which was recovered within six months; two patients had non-obstructive post-surgical vesical instability. No patient had changes in the mechanism of urinary continence. PMID- 9412195 TI - [Vasectomy and arteriosclerosis: are they related?]. AB - There has been a remarkable increase in recent years in the demand by the male population of vasectomy procedures as a sterilization technique. Several recent studies have questioned its safety. We have evaluated in 103 healthy males the effect of vasectomy on plasma lipoprotein concentration, blood pressure and body mass index, all acknowledged risk factors for arteriosclerosis. No statistically significant change was found either in plasma lipoprotein concentration or blood pressure. The only statistically significant (p < 0.05) change was in terms of body mass index, which experiences a mean increase of 1.22%. Our results cannot confirm that vasectomy plays a role as risk factor for arteriosclerosis. PMID- 9412196 TI - [Adenocarcinoma in ileocystoplasty]. AB - Case report of an adenocarcinoma in ileocystoplasty in a 54-year old female that at the age of 24 had undergone right nephrectomy and partial cystectomy surgery followed by vesical extension with ileon due to urinary tuberculosis. Discussion on the rare incidence of this condition, its diagnostic features, the need for long-term follow-up and the relevance of a broad resection of the affected intestinal portion. PMID- 9412197 TI - [Primary adenoma of the female urethra]. AB - Primary melanoma of the female urethra is very rare. Early diagnosis is very difficult and thus involves a long-term non-resolutive therapeutic approach. This paper presents the case of a 61-year old female patient with this type of tumour and nodular dissemination at the time of diagnosis. The patient developed lung and cranial metastasis with 5-year survival. PMID- 9412198 TI - [Paratesticular lipoma]. AB - In spite of being the most frequent benign tumour of the spermatic cord, absolute frequency of this neoplasia is very low. We discuss here a case report of an spermatic cord lipoma with several years evolution. Although ultrasound is useful for clinical diagnosis, exploratory scrototomy and surgical exeresis of the mass is the only reliable diagnosis and viable treatment. PMID- 9412199 TI - [Retroperitoneal cystic mass. Giant exophytic gastric leiomyosarcoma]. AB - Presentation of an unusual case of pre-operatively non-striated retroperitoneal cystic mass confirmed to be a gastric leiomyosarcoma after pathoanatomical study. Considering the rarity of the case, we reviewed the literature as well as the epidemiological, clinical, pathoanatomical and surgical characteristics of gastric leiomyosarcomas which, as in the present case, can appear as retroperitoneal mass. PMID- 9412200 TI - [Cystadenoma of seminal vesicles]. AB - Cystadenoma of the seminal vesicles is a extremely rare pathology. To our knowledge only eight cases have been reported in the literature. We report a new case of this benign tumor, bilaterally located and incidentally found at surgery. Literature is reviewed and clinical, diagnostic and therapeutic aspects of these are discussed. PMID- 9412201 TI - [Infrequent association: massive inguinoscrotal bladder hernia, multiple scrotal lithiasis, and bilateral obstructive uropathy]. AB - Vesical hernias occur in a significant number of patients with inguinal hernia, though massive inguinoscrotal vesical hernias are uncommon. This paper presents a case report of a male patient where massive inguinoscrotal vesical hernia, and scrotum-located multiple vesical lithiasis was associated to a large amount of lithiasic mass and bilateral ureterohydronephrosis. We consider this association of urinary conditions extremely rare, since to our knowledge bilateral ureterohydronephrosis has only been formerly documented in one case. PMID- 9412202 TI - [Retroperitoneal laparoscopic nephrectomy in children]. AB - We present two cases of obstructive uropathy nefrectomy through a retroperitoneal approach was performed. Renal differential function by means of a 2,3 dimercapto succinic acid renal scan showed less than 10% on the ipsilateral kidney to the diagnosis pathology. The size of the kidneys meant no hindrance during its nefrectomy. Morcellation within the organ bag was required for its removal without needing to broaden the 10 mm port opening. The patients were discharged home 48 hours after surgery and they returned to school within the first postoperative week. PMID- 9412203 TI - [Difficult urethral catheterization because of giant inguinal hernia]. PMID- 9412204 TI - [The placebo effect and clinical research in urology]. PMID- 9412205 TI - [Prostatic electrovaporization, a new treatment for prostatic hyperplasia]. AB - TUR remains the choice technique for the surgical management of bph BUT, though effective, is not entirely free of morbidity. Over the last few years, new techniques have been developed trying to decrease morbidity, of which laser procedures are the most successful ones. We report here the prostate electrovaporization technique and our preliminary results in 75 patients seen over a 5-month period. The efficacy of the system is evaluated using the IPSS score and peak flow values; safety was assessed through the complications arisen, vesical catheter time and operating time. Mean hospital stay was 2 days, length of operation 32 minutes and post-operative vesical catheter time 48 hs; at 3 months, mean peak flow was 19.2 ml/seg and mean IPSS 7. Based en our preliminary results, quick convalescence, decreased cost and casiness of the technique, we consider prostate electrovaporization a likely alternative to conventional TUR, although further longer-term studies are warranted in order to evaluate the histological changes caused by this procedure on the prostate tissue. PMID- 9412206 TI - [Treatment of benign hyperplasia of the prostate using thermal transurethral needle ablation (TUNA)]. AB - Transurethral needle ablation of the prostate (TUNA) is a new, fast and minimally invasive device that produce a selective necrosis of the prostatic gland by delivering low level radiofrequency energy. We describe our experience with this new technique. A total of 42 patients suffering from symptomatic BPH were treated with this procedure. The original generator was used in 27 patients (group 1). A new generator allowing a more homogeneous application of intraprostatic temperature was used in 15 patients (group 2). The patients pretreatment evaluation consisted of World Health Organization symptom score and quality of life, digital rectal examination, uroflowmetric parameters, residual volume, transrectal ultrasound and PSA. Follow-up was performed using the same pretreatment parameters at one month, three months, six months and twelve months. All patients were treated using urethral xylocaine with intravenous or intramuscular sedation (petidine clorhidrate) and tolerance was good. IPSS and quality of life decreased significantly in both groups at first month after treatment and kept low up to twelve-month control. Peak flow rate increased from 7.7 +/- 3.7 ml/sec to 10 +/- 4.1 ml/sec at the twelve-month control in group 1 (p > 0.05), and from 7.6 +/- 2 ml/sec to 9.8 +/- 3.3 ml/sec in group 2 (p > 0.05). Residual volume decrease was statistically significant in group 2 (p < 0.05). No significant complications were encountered. Five patients in group 1 and one patient in group 2 required TURP some time in the follow-up (14%). In our experience, after one year of follow-up, improvement in subjective parameters is evident, although uroflowmetric improvement is moderate and with no statistically significance. No differences were found between both groups of treatment. PMID- 9412207 TI - [Comparative analysis of various techniques for the repair of stress urinary incontinence in women. Review of our experience]. AB - OBJECTIVE: We aim to expose our experience in the surgical correction of stress urinary incontinence (SUI) and to analyze the possible factors that could modify the outputs of this type of surgery. METHODS: We have studied 114 women who underwent surgery (60 Raz, 36 Burch, and 18 vaginal wall sling), with a mean follow-up time of 10.5 months. RESULTS: We have not succeeded in demonstrating that preoperative factors such as age, irritative voiding symptoms; history of prior hysterectomy or urethropexia, neurological disease, diabetes or recurrent urinary tract infections; the finding of cistocele, a positive Bonney-Marchetti test or bladder unstability, play any role in the outputs. The failure rate was 16.7% for vaginal wall sling procedure, 35% Raz, and 33.3% Burch. "Survival" analysis did not demonstrate differences related to the procedure or the surgeon. We discovered and important decrease of continence rate with time from the intervention. Higher incidence of postoperative pain, urinary retention and greater residual urine were detected with transvaginal procedures. There was no difference in the incidence of wound infection. CONCLUSIONS: We don't believe that the selection of candidates for this type of surgery should be carried out in base to the above mentioned preoperative factors. Also, we observed a consistent decrease of postoperative continence with time. Finally, we have detected a greater incidence of complications after transvaginal procedures. PMID- 9412208 TI - [Percutaneous treatment of renal cysts with iodinated povidone injection. Long term clinical course]. AB - At present there are several minimally invasive options for the treatment of symptomatic simple renal cysts. One of them, the percutaneous puncture with injection of sclerosant substances, has offered good results. Our study has been conducted in 15 patients with symptomatic simple renal cyst treated by evacuant percutaneous puncture and povidone-iodine injection as sclerosant agent, and makes a short- and long-term evaluation of the results obtained using this procedure. Complete cyst recession with no ultrasound relapse during follow-up was seen in 13 cases (86.5%). Only two patients showed a persistent residual cyst that caused no symptomatology. The easiness of performance, absence of complications and good results obtained make this technique a valid option for the treatment of symptomatic renal cysts. PMID- 9412209 TI - [Cost-effectiveness comparative study of ceftriaxone verus cefotaxime in the treatment of complicated urinary infections]. AB - Prospective, randomized, multicenter study in 267 patients with complicated urinary infection from 9 hospitals nationwide. Drug treatment was either Ceftriaxone 1 g once daily parenterally or Cefotaxime parenteral 1 g 8 hourly for a minimum of 7 days. Patients were clinically, analytically and microbiologically evaluated before and after treatment to assess the efficacy and tolerance of both drug products. To evaluate treatment cost, we used the price of both drugs and the material required for their administration (syringe and disposable needle). 119 patients were excluded from the cost-efficacy evaluation and 148 remained in the study (75 assigned to treatment with Ceftriaxone and 73 to Cefotaxime). Clinical efficacy of treatment was 93% and 87.6% for Ceftriaxone and Cefotaxime respectively (p > 0.05). Cost per patient was 27,347 pesetas for Ceftriaxone and 34,490 for Cefotaxime (p < 0.05). PMID- 9412210 TI - [In vitro study of the process of urinary calculi fragmentation with endoscopy, light microscopy, and scanning electron microscopy]. AB - Stones with different compositions respond differently to shock wave lithofragmentation. Likewise, the various lithotrity systems used may have different effects on the stones. To determine the relationships between stone composition and their fragmentation patterns, we conducted an in vitro study using endoscopy, magnifying glass, light microscope and scanning electron microscope on fragments obtained after lithotrity of 60 pure stone with different compositions: calcium oxalate monohydrate and dihydrate (OXMH and OXDH), phosphocarbonate (PC), ammonium magnesium phosphate (AMF) and uric acid (UA). Fragmentation was carried out with 4 different lithofragmenting sources (electrohydraulic, piezoelectric, ultrasound and pulse laser). No morphologic differences in the fractures induced by the various lithofragmenting sources were demonstrated. OXMH and UA stones basically break up by intercrystalline fracture and splitting of their concentric plates. OXDH breaks up mainly by intercrystalline fractures aided by the fibrillar organic material and phosphocarbonates found in the intercrystalline spaces. Fragmentation in infective stones (AMF and PC) occurs across the intercrystalline surfaces and by intracrystalline fracture. Ammonium urate fragments break up by intracrystalline fractures that run across the equatorial plane of its characteristic acicular microspheres. PMID- 9412211 TI - [Radical cystectomy with orthotopic urinary diversion in patients older than 70 years of age]. AB - Some patients with infiltrant vesical cancer can be treated successfully with radical cysto-prostatectomy and urinary by-pass and increasingly more authors publish successful results in series of selected patients over 70- and 80-year old. Between February 1988 and July 1996, 18 radical cystectomies with orthotopic urinary by-pass were performed in the Urology Service, Policlinico Vigo, in patients over 70 (range 70-84 years), with an operative mortality rate of 11%. 8 patients (44%) developed immediate complications and 9 patients (50%) presented distant complications. Overall mortality in our series was 33.3% and survival 66.6% after a mean follow-up of 30.7 months. We believe orthotopic-continent by pass is a valid alternative with operative mortality and complication rates similar to those of ileal ducts, with the advantage for the patient of avoiding permanent urinary stoma. PMID- 9412212 TI - [Reutilization of a transplanted kidney by a 2nd receptor]. AB - Contribution of what we believe to be the first case of successful reutilization of the same renal graft by a second receptor after functioning for some time in other patient. Transplantectomy was performed 8 days after initial RT, subsequent to the recipient suffering a fatal AVC. The purpose of this paper is to underline the sequence and technical methodology of this procedure which may become a resource to be taken into account in similar cases. PMID- 9412213 TI - [Vaginal metastasis of renal adenocarcinoma: report of 2 clinical cases]. AB - Presentation of 2 case reports of female patients with vaginal metastasis from renal adenocarcinoma presenting genitorrhage which was part of a polymetastatic picture. Discussion of the clinical, diagnostic and prognostic aspects of this disease. PMID- 9412214 TI - [Ultrasonography imaging of the torsion knot in spermatic cord torsion]. AB - Presentation of the finding of the echographic image of a torsion knot as a pathognomonic sign of spermatic cord torsion. Discussion of this finding as well as aspects related to imaging diagnosis of cord's torsion. PMID- 9412215 TI - [Spontaneous bladder perforation secondary to bladder candidiasis]. AB - Presentation of an spontaneous vesical perforation secondary to vesical candidiasis in a patient with in-dwelling catheter due to BPH. Analysis of etiological factors, clinical signs and symptoms, diagnosis and management of both entities. PMID- 9412216 TI - [Intestinal obstruction and peritoneal carcinomatosis after endoscopic resection of transitional carcinoma of urinary bladder]. AB - Gut involvement in bladder tumours is low, even exceptional in the presence of surface, low-grade neoplasia. The authors explain their experience in the diagnosis and management of a patient treated endoscopically for a vesical surface tumour which subsequently exhibited peritoneal and gut metastatic seeding. The various mechanisms for gut dissemination of vesical neoplasias and the repercussion of their endoscopic management are discussed. PMID- 9412217 TI - [Gonadoblastoma in Swyer syndrome]. AB - Swyer's syndrome is a pure gonadal dysgenesis with female external genitalia. A likely complication is the appearance of tumour's dysgenetic gonads. This paper studies a case report of gonadoblastoma in a female patient with this syndrome. PMID- 9412218 TI - [Usefulness of laparoscopy trocars in bladder and perivesical drainage]. AB - Presentation of a new alternative to the traditional percutaneous vesical approach using a laparoscopic trocar. This approach fulfils two purposes: the first one, to reduce the cost of vesical drainage (recyclable material); the second and more interesting one, to avoid draining surgical procedures and uretral lesions during mechanical vesical lithotripsy by using a direct percutaneous access. The difference between this method and the previous ones is not the access route but the materials used. PMID- 9412219 TI - [Pheochromocytoma]. AB - Presentation of 7 case reports of pheochromocytome, diagnosed and treated in our Centre between 1981 and 1995. Clinically all patients had hypertension. Three presented the triple condition of hypertension, pulsatile headache and palpitations. The most useful analytical studies were urine vainillylmandelic acid (VMA) and catecholamines. The main radiologic method was the scanner (CT). Pre-surgical preparation was with alpha-blockers in 5 patients, adding beta blockers in 3. Treatment was surgical in all cases, and the approach was selected based on the tumour's size and location. One patient with severe rheumatic heart disease died on day 3 post-surgery. Mean follow-up is 19 months, and only one patient requires anti-hypertensive medication following surgery. PMID- 9412220 TI - [Renal transplant. Teaching in urology]. PMID- 9412221 TI - [Evolution and prognostic factors of adenocarcinomas of the prostate metastasizing at diagnosis]. AB - OBJECTIVES: In 30-40% patients, prostate adenocarcinoma is diagnosed already in the metastatic phase, a percentage that will depend on the number of patients with localized cancer that we are unable to detect. Hormonal suppression is the most widely accepted therapeutical option, although there are doubts on the value of rescue treatment after the hormone-refractive stage. This paper analyses those parameters as well as the main prognostic factors in a series of 135 patients with metastatic prostate cancer at diagnosis. MATERIAL AND METHODS: Between 1983 and 1996, 414 patients were diagnosed with prostate adenocarcinoma in the Urology Unit. Mostoles Hospital, 135 of which (32.6%) were metastatic at the time of diagnosis and were managed as follows: 113 (84%) were treated with maximum androgenic blockade (MAB). 13 (9.6%) with orchiectomy and antiandrogens, 5 (3.7%) with various treatments, and only 4 received symptomatic treatment. Of those treated with MAB, 97 (72%) continued treatment after the hormone-refractive stage and 16 (12%) were given stramustine phosphate instead of the antiandrogen. Response monitoring was done basically by means of serial PSA determination. The parameters analyzed included survival and the following potential prognostic factors: age, performance status, metastatic bone pain, tumour diagnosis based on number of metastasis, prior PSA level, Gleason, local stage, M1 type at diagnosis based on the 1992 TNM classification, and response to the various treatment applied. RESULTS: Mean age: 72 years. Over an average of 25 (0-127) months, 80 (59%) patients have died; mean follow-up of patients alive at end of study: 24 months (3-111). Lost to follow-up: 6 patients (4.4%). Up to 1991, the proportion of patients with metastasis was 48%; since 1992, 24%. Percentage of patients diagnosed due to clinical manifestations of the metastasis (25 patients) over these two periods increased, mean age decreased and the proportion of patients with highly aggressive tumours increased. Mean overall survival, 26 months: influential prognostic factors: diagnosis due to metastasis and Gleason greater than 7; very poor prognosis for those receiving no hormonal therapy, with no differences between drug versus surgical treatment. Tumour-dependent mean survival, 32 months; influential prognostic factors: performance status, metastatic bone pain, diagnosis due to metastasis and Gleason greater than 7; very poor prognosis for those receiving no hormonal therapy. Progression-free interval, 19 months; influential prognostic factors: metastatic bone pain, PSA higher or lower than 90. Gleason greater than 7 and local stage: no differences between treatments. Mean survival after progression, 6 months; influential prognostic factors: diagnosis due to metastasis, M1b versus M1c patients: increased survival in patients rescued with stramustine phosphate. CONCLUSIONS: The proportion of prostate adenocarcinomas with metastasis at diagnosis shows a trend to decrease, although the percentage of patients who are diagnosed by the sings and symptoms of their metastasis is increasing. These patients should be treated with pharmacological or surgical hormone-suppression. Rescue treatment with stramustine phosphate prolongs survival. Influential prognostic factors: Gleason greater than 7, metastatic bone pain, tumour extent and previous PSA. PMID- 9412222 TI - [Usefulness of computed tomography for the staging and management of post traumatic renal lesions]. AB - OBJECTIVE: Use of computerized tomography (CT) criteria for the staging of 50 cases of renal trauma diagnosed in the Hospital General Universitario "Gregorio Maranon" over the 1989-1995 period. CT is used as the choice technique to diagnose renal lesions of traumatic etiology in patients haemodynamically stable. MATERIAL AND METHODS: Retrospective study of 50 patients (age range: 16-73 years; sex distribution: 39 male/11 female) with a history of abdominal trauma and CT confirmed renal injury. Following Hodges-Federle criteria for patients with no underlying renal disease (n = 44), lesions are staged into 4 categories or grades according to CT findings. The traumatic mechanism is correlated to clinical semiology, staging and associated extrarenal lesions. Also, traumatic renal lesions in patients with previously diseased kidneys (n = 6) are analyzed separately. RESULTS AND CONCLUSIONS: Percentile distribution of lesions was as follows: grade I: 50% (n = 22), grade II: 29.5% (n = 13): grade III: 11.4% (n = 5): grade IV: 9.1% (n = 4). Compared to others, our series shows a decreased percentage of mild or grade I lesions. It is confirmed that CT is a very valuable imaging method for an adequate staging of post-traumatic renal lesions, the diagnosis of associated extra-renal lesions and, therefore, for the subsequent therapeutical management. PMID- 9412223 TI - [Results of the conservative treatment of severe renal trauma in childhood]. AB - Surgical treatment of severe renal trauma usually ends in loss of high percentage of kidneys. In consequence of this, in the last decade several authors prefer a conservative management of kidneys severely injured. There include stabilization in Intensive Care Unit, with the goal of preservation the most possible functioning renal tissue. The purpose of this paper is to present the results of conservative management of severe renal trauma in our Center in the last five years. We conclude that this type of management is accurate and effective. PMID- 9412224 TI - [Renal hematoma after shockwave extracorporeal lithotripsy]. AB - OBJECTIVE: Renal haematomas after shock wave extracorporeal lithotripsy (SWEL) represent a potentially serious complication. This paper examines those cases of post-SWEL renal haematoma seen in our Centre, analyzing the likely risk factors. PATIENTS AND METHODS: Between May 1988 and June 1996, 12,800 patients were treated with 15100 lithiasis at some level of the urinary tract requiring 16,000 SWEL sessions. All treatments were done with a Dornier HM-4 lithotripter. Voltage applied ranged from 18 to 26 Kv, averaging 2500 waves/session. Complementary testing (ultrasound/computerised tomography) was requested immediately after treatment if clinical complications were suspected. RESULTS: A total of 10 renal haematomas (0.078%) were diagnosed. Six cases were mild, but 4 presented extensive haematoma with significant haemodynamic consequence. Although in one case nephrectomy was undertaken to control haemorrhage, death finally occurred by disseminated intravascular coagulation. Four patients who developed haematoma were hypertensive and 3 had a previously corrected haemostasis alteration. CONCLUSIONS: The possibility of renal haematoma should be taken into account in the face of persistent and unjustified pain after SWEL treatment. Normalization of blood pressure values, correction of urinary infection as well as adequate correction of haemostatic disorders is advisable. PMID- 9412225 TI - [Short hospital stay in the treatment of benign prostatic hyperplasia using transurethral resection]. AB - INTRODUCTION: Prostate TUR is the most common approach used in the surgical management of BPH. The highest expenditure portion of this treatment occurs during the post-operative. PATIENTS AND METHOD: 42 cases of prostate TUR were analyzed by GRD type, prostate weight, hospital stay and complications in the peri- and post-operative periods. Both the approach used and the degree of patient's satisfaction are analyzed. RESULTS: Mean prostate weight is 46 g (range 23-69). 45% cases involve GRD 336 and 55% GRD 337. Mean hospital stay for the procedure was 2.5 days. 2.3% of the series developed long-lasting temperature, 2.3% required transfusion and 2.3% a second intervention. Around 81% patients are satisfied with the results. CONCLUSIONS: It is possible to reduce costs by reducing hospital stay without impairing care quality and patient satisfaction. PMID- 9412226 TI - [Experience of the Valdivia Hospital in renal transplant: an analysis of our first 100 cases]. AB - From September 1980 until October 1995 we have performed 100 renal transplant on 95 patients. Up to May of 1996 the medium follow-up was of 69.7 months (in a range of: 15 to 196 months). Seventy-nine kidney implants were performed from related living donors and 21 from dead donors. The average age of recipients was 35.8 years (SD: +/- 12.5 years). Chronic renal insufficiency of recipients is reviewed as well as their accompanying pathology. We found that in a 20% of the cases the donor's kidney had multiple renal arteries. In most of the ureteral vesical implantations were performed with Salvatierra's technique. Preservation cold time for the cadaveric donor kidneys was over 24 hours. In only a 33% of the cases Cyclosporine was used as the immunosuppression therapy. The 9% of ureteral complications are outlined. TBC is an important cause of complications and death of patients. To date there are 76 functioning kidney allografts and 14 lost graft kidneys. Ten patients have died, 3 of them with functional graft kidney. Actuarial graft survival rate for 1, 5 and 10 years are 92.7%, 78.7% and 59.3% respectively. PMID- 9412227 TI - [Chronic orchialgia. A diagnostic and therapeutic hypothesis]. AB - OBJECTIVE: The objective of this study is to offer patients with chronic testicular pain and no physico-pathological finding, a likely cause not usually taken into account such as uricemia. The hypothesis of an intracanalicular deposit of uric crystals and/or the resulting alteration in nerve endings is suggested. METHOD: The study involved a total of 60 patients with chronic orchialgia and no detectable testicular pathology seen over a 5-year period. Mean age 35.6 years. They were all evaluated with routine laboratory tests, uricemia, uricosuria and in specific cases uroculture and Stamey's method. Orchidometry, vascular doppler and ultrasound were used in all cases. RESULTS: Presence of hyperuricemia was corroborated in 61.6% cases. Based on figures found, patients were divided into three groups. Patients were treated with a low-purine diet plus Allopurinol 300 mg/day, and symptoms receded in 81.06% cases. CONCLUSIONS: Due to the high incidence of hyperuricemia and the significant symptomatological relief obtained with specific treatment, we believe its determination should become routine. PMID- 9412228 TI - [Agenesis of vas deferens and cystic fibrosis]. AB - Congenital bilateral absence of vasa deferens appears in 6% of obstructive azoospermia, and 60-70% of these patients also have cystic fibrosis mutations. Unilateral aplasia or agenesia of vasa deferens occurs in less than 1% male individuals and some studies have found that up to 43% cases show mutations in the cystic fibrosis gen. We contribute four case reports of bilateral agenesia who were seen for infertility, all of which showed presence of mutation. In none of the two cases of unilateral agenesia, who consulted for vasectomy, a mutation in the cystic fibrosis gen was found. Patients with bilateral agenesia and their partners should be screened for cystic fibrosis, prior to spermatic microaspiration and assisted fecundation. PMID- 9412229 TI - [Primary non-hodgkin lymphoma of the testicle]. PMID- 9412230 TI - [Primary lymphoma of the testicle]. AB - Case report of a testicular lymphoma. A diagnosis of primary neoplasia of the testis was made since no involvement to the lymphoreticular system in any other organ was demonstrated. We review the clinical, diagnostic and therapeutical aspects and make a note on the rarity of our case, a type T tumour, due to the exceptional nature of this variety within primary lymphomas of the testis. PMID- 9412231 TI - [Vesical endometriosis after cesarean section: diagnostico-therapeutic aspects]. AB - Endometriosis is a benign condition with an aggressive behaviour defined by the presence of ectopic endometrial tissue, outside the uterus. It occurs in 15-20% women with child bearing potential. Most commonly it affects organs such as the ovaries, uterine ligaments, fallopian tubes, rectum and the cervico-vaginal region. Involvement of the urinary tract, however, is rare. It can be seen in just about 1% cases, vesical location being the most frequent of these presentations (84% cases). We describe one case of vesical endometriosis that developed after a cesarean section. The intra-operative findings confirmed the existence of infiltration of the detrusor muscle and the vesical mucosa by endometrial tissue from the area of the uterine incision. A discussion of the different diagnostic and therapeutic options is also included. PMID- 9412232 TI - [Congenital anterior urethral diverticulum]. AB - Contribution of one case report of congenital urethral diverticulum, located in the anterior urethra with narrow opening, that occurred in an adult male patient as a post-traumatic urethrorrage and presented with no obstructive symptomatology. The occasional incidence, etiology and atypical presentation form of this entity are examined. The diagnostic features, where urethrocystography still prevails, as well as the surgical alternatives are also discussed. PMID- 9412233 TI - [Resolution with vascular endoprosthesis after first angioplasty failure in the treatment of post-transplant stenosis of the renal artery]. AB - We report a case of renal artery stenosis after a living transplant kidney, treated successfully with percutaneous transluminal angioplasty (PTA) and vascular endoprosthesis. PTA is the initial treatment of choice for most patients with high grade transplant renal artery stenosis. Surgical revascularization is indicated if PTA cannot be done or is unsuccessful. PMID- 9412234 TI - [Verruciform xanthoma of the penis]. AB - Verruciform xanthoma is a rare benign lesion. The most common presentation is at the oral mucosa level, although it has also been described at other locations. Our case is the twelfth verruciform xanthoma of the penis ever published. We highlight the relevance of the differential diagnosis and its excellent prognosis, the choice treatment being the simple exeresis of the lesion. PMID- 9412236 TI - [Cysts of the median raphe (prepuce and perineum). Brief contribution]. AB - A few remarks on the clinical records of two children with middle raphe cysts located in the pre-putium and the perineum. For a better understanding of this entity, the organogenetic dynamics of the penis, scrotum, pre-putium and male urethra are outlined and redefined. Middle raphe cysts are tegumentary formations that arise as a result of "tissue trappings". This entity should not be suggestive of malignancies. Therapeutical restraint should be the rule. Exeresis should only be indicated in the face of a hypothetical complication (e.g., infection, large volumes, etc.) or because of aesthetic reasons. PMID- 9412235 TI - [Cysts of median raphe]. AB - Middle raphe cysts are very uncommon lesions in the daily clinical practice and can go unnoticed based on their size. We contribute two cases where a quick growth is seen over the last few months. Their histological origin is discussed using specific staining and immunohistochemical techniques. PMID- 9412238 TI - Proceedings of the 5th Washington Symposium on Dietary Fiber: The Vahouny Fiber Symposium. Washington, DC, March 26-29, 1996. PMID- 9412237 TI - Proceedings of the American Institute for Cancer Research's 7th annual conference on dietary fat and cancer: genetic and molecular interactions. Washington, D.C., August 28-30, 1996. PMID- 9412239 TI - ACS updates Guidelines on Screening for Colorectal Cancer. PMID- 9412240 TI - AUA releases Treatment Guidelines for Female Stress Urinary Incontinence. PMID- 9412241 TI - New Approaches to Cardiovascular Therapy. Proceedings of a symposium. Anaheim, California, March 15, 1997. PMID- 9412242 TI - Beta Blockade in Cardiovascular Disease: Current Perspectives on Patient Protection. Symposium proceedings. New Orleans, Louisiana, November 10, 1996. PMID- 9412243 TI - Bulimia outcome. PMID- 9412244 TI - Isaac Max Rubinow. PMID- 9412245 TI - [Neurobiology of awakening and wakefulness]. PMID- 9412246 TI - [Adaptation of the adult brain to denervation: reorganization of connexions and neural maps]. PMID- 9412247 TI - [A new form of infectious action. Prions and spongiform encephalopathies]. PMID- 9412248 TI - [Evolutionary theory of knowledge]. PMID- 9412249 TI - [Laparoscopic surgery. Critical view]. PMID- 9412250 TI - [Influence of biotechnology and genetic engineering in medical therapy]. PMID- 9412251 TI - [Current concepts on the correlation of endometriosis and sterility]. PMID- 9412252 TI - [Positron-emission tomography in oncology]. PMID- 9412254 TI - [Thoracic epidural anesthesia: still controversial?]. PMID- 9412253 TI - [Clinical applications of positron-emission tomography in brain pathology]. PMID- 9412255 TI - [Thoracic epidural anesthesia--more than an anesthesia technique]. AB - Thoracic epidural anaesthesia (TEA) faces growing interest as an adjuvant anaesthetic and postoperative analgesic regimen. The procedure allows a specific blockade of nociceptive reflex arches and may exert beneficial effects on stress induced alterations of organ function. Myocardial blood flow to areas at risk is improved, and paradoxical reactions of atherosclerotic coronary arteries after sympathetic stimulation are suppressed. After cardiac surgery, TEA improved postoperative recovery and resulted in better haemodynamic stability and allowed earlier extubation. During vascular surgery, the graft occlusion rate was significantly decreased. The improved pulmonary function after TEA is due to superior pain relief which allows the patients to breathe and cough sufficiently. After upper abdominal surgery, TEA leads to improved recovery of gastrointestinal function which reduces the risk of bacterial translocation. Although lumbar epidural anaesthesia is preferred by many anaesthesiologists as there is no risk of traumatizing the spinal cord, many positive effects are forgone. With insufficient rostral spread of a lumbar epidural block above the fifth thoracic level, cardiac complications can occur due to reflex activation of sympathetic outflow in unblocked thoracic regions. When the contraindications are carefully observed, TEA can be safely performed in most patients. PMID- 9412256 TI - [Preoperative hemodilution with bovine hemoglobin. Acute hemodynamic effects in liver surgery patients ]. AB - Haemoglobin solutions can be an alternative to allogeneic red-cell transfusions because they combine colloid osmotic with oxygen transport properties. Since severe toxic side effects have been overcome by ultrapurification, clinical interest has been focused on haemodynamics changes during application of haemoglobin preparations. The present clinical study examines changes of haemodynamic and oxygen transport parameters during and after haemodilution with ultrapurified polymerized bovine haemoglobin (HBOC-201) in comparison to hydroxyethyl starch (HES). METHODS: After approval of the Ethics Committee, 12 patients (6 males and 6 females, mean age 59 +/- 10 years, ASA 1-2) undergoing elective liver resection were randomly allocated to receive either 3 ml.kg-1 6% HES 70,000/0.5 (group 1) or 0.4 g.kg-1 HBOC-201 (group 2) within 30 min following autologous blood donation of 1 l and substitution with 2 l Ringer's lactate. Measurements of blood gases, haemodynamics, and oxygen transport parameters were performed after induction of general anaesthesia, prior to and after blood donation, during and after infusion, at the beginning of surgery, and in the intensive care unit. RESULTS: Demographic characteristics did not differ between groups. In contrast to the HES group, mean arterial pressure increased by 18% over baseline measurements in group 2. While pulmonary vascular resistance showed a trend to higher values in group 2, systemic vascular resistance increased to a maximum of 42% over baseline in group 2 and was twice as high as in the HES group. The cardiac index was lower in the HBOC-201 group than in the HES group. During and after HBOC-201 infusion, mixed-venous oxygen saturation and content and calculated oxygen delivery were lower in group 2 in comparison to group 1, while the oxygen extraction ratio was higher in group 2. Free haemoglobin reached a maximal concentration of 1.0 +/- 0.2 g.dl-1 30 min after the HBOC-201 infusion was started, but was not detectable in urine over time. The mean intravascular half-life of HBOC-201 was 8.5 h. CONCLUSIONS: Patients did not show any severe complications during and after infusion of HBOC-201. However, vasoconstrictive side effects resulted in increased systemic but not pulmonary resistance. Ongoing studies with higher doses of HBOC-201 applied in a larger number of patients will probably reveal potential clinical consequences of the demonstrated haemodynamic changes. PMID- 9412257 TI - [The effects of dopexamine. Transpulmonary shunt volume in thoracic surgical procedures with one-lung respiration]. AB - OBJECTIVE: To study the influence of dopexamine on pulmonary shunt and hypoxic pulmonary vasoconstriction during major thoracic surgery with one-lung ventilation (OLV). DESIGN: Prospective, randomised, placebo-controlled study. SETTING: University hospital. PATIENTS: Twenty adult patients undergoing elective pulmonary resection. ANAESTHESIA: General anaesthesia was performed using propofol, fentanyl, N2O and vecuronium. Volume-controlled ventilation was performed to maintain normocapnia over the whole investigation period. During OLV, the tidal volume was reduced and the respiratory rate was increased to avoid a peak airway pressure exceeding 40 cm H2O. Furthermore the FiO2 was increased to 1.0 and the external PEEP was removed during OLV. INTERVENTIONS: The patients received either dopexamine at 2 micrograms/kg/min (group A, n = 10) or 0.9% saline as control (group B, n = 10) after assessing the baseline values. MEASUREMENT AND RESULTS: The following cardiorespiratory variables were recorded: Heart rate, mean arterial pressure and mean pulmonary arterial pressure. Cardiac output was measured by thermodilution using a continuous cardiac output thermodilution catheter. Arterial and mixed venous blood gas analysis were measured from simultaneously drawn samples. Cardiac index (CI), systemic vascular resistance index, pulmonary vascular resistance index, oxygen delivery index (DO2I), oxygen consumption index and the venous admixture were calculated using standard formula. Furthermore, pressure-flow-curves were constructed to analyse flow independent changes in the pulmonary vascular resistance. Data were recorded at the following times: After induction of anaesthesia in stable haemodynamics during two-lung ventilation (baseline values, T0), intraoperatively during one lung ventilation (T1) and postoperatively after re-establishing two-lung ventilation (T2). Patients characteristics, data from the preoperative lung function testing and surgical procedures did not differ significantly between the groups. CI increased in the dopexamine group from 2.5 +/- 1.2 1.min-1.m-2 (T0) to 3.6 +/- 0.9 l.min-1.m-2 (T1) and 4.0 +/- 1.3 l.min-1.m-2 (T2). The course of the intrapulmonary right-to-left shunting did not differ between the groups. In the dopexamine-treated group the DO2I increased from 430 +/- 143 ml.min.m-2 (T0) to 652 +/- 255 ml.min.m-2 (T1) and 653 +/- 207 ml.min.m-2 (T2). Regarding the pressure-flow-curves there was no difference during OLV between the two groups indicating no major blocking effect of dopexamine on hypoxic pulmonary vasoconstriction. CONCLUSION: It is concluded that dopexamine can be used to improve haemodynamics and oxygen delivery during thoracic surgery without increasing venous admixture during one-lung ventilation. PMID- 9412258 TI - [Propofol and postoperative nausea and/or vomiting]. AB - The objective of this prospective, randomised study was to investigate the incidence of postoperative nausea and/or vomiting (PONV) during the first 24 h postoperatively. For a quality assurance study on PONV, we compared two established general anaesthetic procedures in 239 patients undergoing four different types of surgery (subtotal thyroidectomy, laparotomy for gynaecological procedures, laparoscopy, and surgery for extra-abdominal procedures). METHODS: All eligible patients provided informed consent. For premedication temazepam 10 20 mg was administered orally. We used propofol (1.5-2 mg/kg) for induction of anaesthesia in all patients, followed by 0.1-0.3 mg fentanyl, 2.5-5 mg droperidol, and for muscular relaxation atracurium or pancuronium according to body weight. Maintenance of general anaesthesia in group A was by administration of isoflurane in a maximum concentration of 0.6 vol.% in 70% nitrous oxide and 30% oxygen and in group B by continuous infusion of propofol (5-8 mg/kg.h) and normoventilation with oxygen in air (Fi02 = 0.3). In both groups additional analgesia was provided intraoperatively by equal dosages of fentanyl up to a maximum of 0.6 mg and clonidine up to 200 micrograms. Episodes of PONV were registered following extubation, during the first 4 h postoperatively, during the period 4-24 h postoperatively, and after the first mobilisation. Pain scores were recorded with the aid of a visual analogues scale. The statistical evaluation was performed using the chi-square or Wilcoxon test. RESULTS AND DISCUSSION: In patients undergoing thyroidectomy or laparotomy, continuous infusion of propofol drastically reduced the incidence of PONV in the first 24 h postoperatively, particularly during the first 4 h (25/41 vs 10/41, resp. 20/32 vs 11/31). The overall incidence of PONV was higher in the first half of the menstrual cycles decreased with patient age, increased with the duration of anaesthesia, and was higher in patients with a history of motion sickness. With the same level of analgesia in both groups, the differences disappeared in the further postoperative course. The use of similar dosages of opioids for pain control in these groups might explain this observation. PONV occurred extremely rarely in patients undergoing laparoscopy (1 resp. 2 of 34), and in those undergoing surgery for extra-abdominal procedures did not occur at all. The explanation may be that the induction of anaesthesia with propofol was followed only by a relatively short duration of general anaesthesia for these surgical procedures, and postoperative pain control was performed solely with non-opioids. CONCLUSIONS: We found that the antiemetic effect of propofol was considerable in the early postoperative period. The higher cost of propofol as compared to other induction agents can be covered by not using nitrous oxide for maintenance of anaesthesia and by the decreased need for antiemetic drugs postoperatively. According to the calculations of our clinical pharmacy, the costs of the propofol infusion regimen exceeded those of balanced anaesthesia by 8.50 DM/h; the need for antiemetics was one-half that of the non-propofol group. Considering a cost of 16 DM for cleaning the bed after vomiting, improvement of the patient's condition during the postoperative period can be achieved without additional expense. PMID- 9412259 TI - [Intraoperative diagnosis of a multilocal pheochromocytoma]. AB - Pheochromocytomas are functionally active, catecholamine-secreting tumours of chromaffin tissue. The mainstay of pharmacological therapy is preoperative treatment with oral phenoxybenzamine. This drug irreversibly alkylates alpha-1 adrenergic receptors on vascular smooth muscle and renders them nonfunctional, thereby causing vasodilatation. The duration of action of a single dose is approximately 24 h. Therefore, postoperative hypotension is a hazard of therapy with phenoxybenzamine if adequate plasma volume repletion is not provided. Prazosin, a short-acting, competitive alpha-1 blocker, has been used preoperatively, but has been criticized for its failure to adequately prevent perioperative hypertensive episodes. We report the case of a 73-year-old woman who was admitted for elective pheochromocytoma resection. Preoperative therapy with phenoxybenzamine was impossible because of the patient's refusal to take the drug. Preoperative antihypertensive preparation was therefore performed with prazosin 30 mg/24 h and metoprolol 100 mg/24 h. During the surgical preparation of the tumor, sodium nitroprusside was started at an average infusion rate of 4.1 micrograms/kg/min. After resection of the primary tumor, when the sodium nitroprusside infusion was stopped the patient exhibited an increase in systolic blood pressure (BP) up to 210 mg Hg. This hypertensive crisis was managed with sodium nitroprusside, nitroglycerin, and esmolol. A multilocular pheochromocytoma was diagnosed. Further stimuli due to tumour palpation resulted in repeated increases in BP. In this manner, two additional areas of tumour could be diagnosed by BP peaks after reduction of the sodium nitroprusside infusion. After complete resection of a total of three tumours, no further hypertensive crises occurred. The patient's postoperative course was uneventful. We conclude that in this patient presenting with an unsuspected multilocular pheochromocytoma, the lack of permanent alpha-blockade was probably helpful in allowing complete resection of all the tumours. PMID- 9412260 TI - [Quality management in emergency medicine]. AB - Although the need for the implementation of a quality management concept for the German emergency medical system (EMS) has been discussed for more than 10 years, such a concept has not been realised on a broad scale. Standardised national data sheets were developed many years ago. They are used by many local agencies, but a data-gathering system on a state or national basis is still lacking. In times of reduced funds for health care expenditures, quality management could be a reliable way to ensure that the EMS provides safe services to the patient based on the current state of medical science in an efficient manner. Based on clear definitions, structure, process, and outcome quality can be analysed, and the results provide the basis for continuous quality-improvement strategies. As not all aspects of the system can be analysed continuously, one has to select areas of special importance. External and internal quality control are equally important. Quality control works on the basis that all EMS team members are motivated to perform on a professional level to ensure that each patient is treated adequately. It evaluates the system to create circumstances that enhance the achievement of this goal. Quality management is not only concerned with mishaps, because areas with documented good performance also provide important information. PMID- 9412262 TI - [The PTT--limit for regional anesthesia?]. PMID- 9412261 TI - [Functional failure of an inspiratory Draeger-circuit system. An example of the fundamental problem of borderline damage to a component part in anesthesia and intensive care]. AB - Unnoticed, minor damage to the unidirectional respiratory valves of the Draeger respiratory circuit may lead to intermittent and unpredictable malfunction, resulting in rebreathing and hypercapnia. The damage may be so minor that normal visual and functional test routines may be insufficient to detect it. We report one case of a potential life-threatening malfunction of the inspiratory valve and also propose economical solutions utilizing altered construction, modified machine-check procedures, or a simple instrument that adds only one step to the machine-check procedure. The general problem of minor but functionally important damage to parts of ventilatory equipment--so-called borderline damage--may not be limited to this particular model or manufacturer. Most users of ventilatory equipment believe that equipment that goes through normal check procedures is either fully functional or nonfunctional. In reality, this is not the case. Intermittent malfunctions due to slightly damaged equipment may be missed with normal machine-check procedures. This problem results in a significant but incalculable increased in risk to patients. Because of unclear reproduceability of intermittent malfunctions caused by borderline damage, there also is an increased forensic risk for the anaesthesiologist. The risk of mechanical malfunction might be displaced by software problems in new-generation ventilators in the market. PMID- 9412263 TI - [Muscle relaxants and the heart muscle]. PMID- 9412264 TI - [Epidural anesthesia for cesarean section]. PMID- 9412265 TI - [Hemostasis and anesthesia. 5th Heildelberg Symposium. June, 20-21, 1997]. PMID- 9412267 TI - [Postoperative pain]. PMID- 9412266 TI - [Desflurane and sevoflurane. An interim balance]. PMID- 9412268 TI - [The legal obligation for postoperative pain therapy]. AB - Postoperative pain therapy, i.e., the symptomatic treatment of acute post surgical pain, is an interdisciplinary obligation of the anaesthesiologist and managing surgeon toward the patient. The failure to provide appropriate pain therapy, in particular withholding analgesic agents, can be regarded as malpractice and result in civil, criminal, and professional legal consequences. As with other medical treatments, the patient must be adequately informed about pertinent details prior to the use of pain therapy, particularly in regard to specific risks and feasible alternative measures. All pain therapy employed must be duly recorded. PMID- 9412269 TI - [The quality of postoperative pain therapy]. AB - Many patients still suffer from unnecessary pain in the postoperative period. Waiting for new technologies or drugs will not improve the status of acute pain management. The establishment of an acute service is needed to reduce the incidence of postoperative pain. Acute pain therapy is one of the great challenges we have to take up. PMID- 9412270 TI - [Neurophysiological aspects of pain and its consequences for the anesthetist]. AB - Nociception is a protective system of the body which prevents it from injury and tissue damage. Human beings respond to noxious stimuli by moving away. They learn by pain to avoid these situations in future. Shortly after major injury, there is a limited analgesic period allowing the body to flee the area of danger, later on, emerging pain compels the body to rest and supports recuperation. While acute pain has a certain meaning, chronic pain does not. It induces a comprehensive suffering including loss of initiative, appetite and vigilance. It reduces life quality, often accompanied by depressive moods. Acute pain causes changes in the central nervous system leading to an increased sensitivity of nociception (hyperalgesia). During healing, the central processing of noxious stimuli is normalised taking minutes to weeks. Sometimes, unknown factors initiate chronification of pain. Changes on a molecular level in peripheral tissue as well as in the central nervous system induce "cellular early genes", a synthesis of c fos, c-jun and other proteins favouring the chronification of pain. All efforts have to be made to depress or interrupt such a development. One of the first steps to pain prophylaxis in a hospital is an optimal surgical technique: incision, extension, limited tissue damage and minimal invasive surgery should guarantee the smallest impairment of the nociceptive system possible. However, nociceptive input is intense and of long duration and leads to central sensibilisation. Postoperative pain has lost its function as surgery anticipates healing. Pain induces a reduction of ventilation, circulation, digestion and increases the risk of other disorders. There is need of aggressive pain treatment for humanitarian reasons and for reasons of late sequelae like permanent pain and increased reduction of function. This is of pivotal importance in patients with amputations or sympathetic reflex dystrophy (SRD). Antinociception is best provided by regional anaesthesia technique with a combination of local anaesthetics and opioids which results in better outcome. Hence, regional anaesthesia techniques are strongly indicated in those patients. Good antinociception may be even more important than it is assumed today. Anand demonstrated a lower morbidity and mortality in 45 newborns undergoing cardiothoracic surgery, when general anaesthesia was performed with high-dose sufentanil versus halothane supplementary doses of morphine. Anaesthesiologists have to reconsider the quality of general anaesthesia: the antinociception of their regimen. PMID- 9412271 TI - [The pharmacologic basis of postoperative pain therapy. Epidural opioid administration]. AB - The administration of epidural opioids is alternatively used in the management of postoperative analgesia. However, the administration is associated with side effects, including respiratory depression, somnolence and pruritus. A rational opioid selection between the hydrophilic and lipophilic opioids morphine, hydromorphone, alfentanil, fentanyl and sufentanil is discussed in this mini review. Thus, the administration of the lipophilic opioid sufentanil might has some advantages. Notwithstanding, epidural opioid administration alone offers no marked clinical advantages compared to the intravenous route. In future, reduced doses of lipophilic opioids and local anaesthetics like bupivacaine 0.05-0.1% may provide benefits over the use of either drug alone and may offer marked clinical advantages over the intravenous route of opioids alone. The same holds true for alpha 2-adrenoceptor agonists as adjuvants. However, multicenter dose-ranging studies are necessary to determine both the ideal concentrations of the drug combinations and the general outcome. Moreover, we must also determine cost effectiveness for our postoperative analgesic techniques. PMID- 9412272 TI - [Obstetrical pain therapy]. AB - Until today, the use of epidural analgesia in obstetrics still remains controversial. In the opinion of many obstetricians the use of an epidural for a healthy laboring parturient is not necessary and can lead to potentially harmful side effects. However, painful labor leads to a maternal stress reaction with the release of epinephrine and norepinephrine. The resulting vasoconstriction of uterine arteries can induce fetal asphyxia in an otherwise healthy fetus or can aggravate asphyxia in already compromised fetuses. Several studies show that epidural analgesia can attenuate the maternal stress reaction and thereby improve maternal and fetal well-being, as long as precautions are taken. The avoidance of maternal hypotension with sufficient volume preload with lactated Ringer's solution or colloids, and decreasing the concentration of local anaesthetics by adding opioids will prevent an increase in instrumental deliveries. With the use of patient-controlled epidural analgesia (PCEA) the amount of local anaesthetics can even further be reduced. PMID- 9412273 TI - [Patient-controlled postoperative epidural analgesia. Prospective study of 1799 patients]. AB - Side effects of postoperative epidural analgesia can be controlled by two strategies: Insertion of catheters into the center of the affected spinal segments and coadministration of local anesthetics and opioids. Both techniques will reduce single drug dosage. Additionally synergistic effects will result in excellent analgesia and the risk of side effects and complications will be minimized. METHODS: Between september 1995 and february 1997 the pain-service of the Klinik and Poliklinik fur Anasthesiologie und operative Intensivmedizin der Westfalischen Wilhelms-Universitat Munster has used this regimen to treat 1799 postoperative patients with patient-controlled epidural analgesia. All patients received an infusion of bupivacaine 0.175%, which was combined with sufentanil 1 microgram/ml in adults under the age of 70 an in children with a body weight > 30 kg. The infusion was adjusted to the individual needs of the patients by a visual analogue scale (VAS-scale: 1 = no pain; 10 = worst pain possible). Analgesia was adequate if VAS-scores were < 4 during rest and < 7 during movement and coughing. The continuous drug administration was combined with additional patient controlled bolus doses. Postoperatively a special observation period to monitor side effects of epidural sufentanil was not defined. All patients were admitted to wards as soon as they fulfilled common criteria for discharge from the recovery room. RESULTS: Mean VAS-scores during the postoperative observation period were within the prior defined limits. On the morning after surgery, however, a reduction in pain relief was observed and analgesia on the first postoperative day could significantly be improved after a 24-h on call pain service has been introduced. Except urinary retention side effects are rare. Probability of motor-blockade is significantly lower in patients with thoracic compared to patients with lumbar catheters. Not any patient suffered from severe complications such as sedation or respiratory depression.de PMID- 9412275 TI - [Risks and complications following peridural anesthesia]. AB - Postoperative neurological sequelae in patients that have received epidural anaesthesia are not necessarily caused by the epidural anaesthetic technique. As a whole, adverse neurological outcomes following epidural anaesthesia may be subdivided into 3 different ethiological categories. A first category involves events that are not at all caused by the epidural, but merely due to the interference of anaesthesia and/or surgery with a preexisting medical condition. A second category includes mishaps such as backache, arachnoiditis, and post dural puncture headache that are solely due to the epidural anaesthesia. Finally, epidural anaesthesia may be a contributory factor in the development of post anaesthetic complications attributable to a pre-existing medical condition that are triggered by anaesthesia, surgery or childbirth. These complications include some of the most dramatic sequelae of major neuraxial blockade, such as spinal epidural abscess, spinal infarction, and spinal hematoma. Although extremely rare, the latter complications often result in permanent major neurological deficits. The present manuscript is a review of the most recent, literature addressing post-anaesthetic sequelae, and will discuss their incidence, pathophysiology, clinical course, diagnosis, prevention, and treatment. PMID- 9412274 TI - [Intravenous versus thoracic-epidural patient-controlled analgesia following extended abdominal or thoracic surgery]. AB - BACKGROUND: Intravenous patient-controlled analgesia (PCA-i.v.) has has markedly improved postoperative pain-relief. Alternatively, peridural anesthesia has been used successfully in high risk patients with the disadvantage of a more intense postoperative care. In this study we compared the applicability of intravenous vs. peridural patient-controlled analgesia on a general ward. METHODS: In a prospective double blinded study 40 patients were randomized after extensive thoracic or abdominal surgery in two groups and received either intravenous PCA (n = 20) or epidural PCA (n = 20). Postoperative monitoring was performed on the general ward by specifically trained nurses. Physiological data, neurological status, the effects of the analgesia and complications were registered before and 48 hours after surgery. Pain intensity was determined by using the visual analog scale (VAS). For the evaluation of wellness and cognitive efficacy psychological tests were performed. RESULTS: Our results show that epidural PCA without administration of a basal rate is a safe method and can be performed on a general ward. Relevant postoperative complications or negative side effects were not registered in both groups. Sufficient analgesia was achieved with both methods. Patients treated with PCA-PDK had a significantly better score regarding vigilance and subjective wellness when compared to patients in the PCA-i.v. group. CONCLUSION: This study demonstrates that epidural PCA can be used on a general surgical ward as an alternative method compared to intravenous PCA. PCA PDK may be advantageous over intravenous PCA since both techniques require similar intense monitoring and side effects in the PCA-PDK group appear to be less. PMID- 9412276 TI - [Quality control in multimodal postoperative therapy]. AB - Effects of anaesthesia and analgesia on postoperative morbidity and mortality remain controversial. Numerous studies have demonstrated that epidural anaesthesia and pain relief by epidural analgesia reduces perioperative stress responses and thus may reduce postoperative morbidity and mortality. In patients undergoing vascular surgery, epidural anaesthesia diminished postoperative hypercoagulability. These patients may benefit from less thromboembolic complications as well as a reduced risk of a re-operation. However, regional anaesthesia does not affect cardiopulmonary morbidity or overall mortality significantly in most clinical studies. One reason for this disappointing finding may be the missing integration of improved postoperative pain relief into general surgical care. A multimodal therapeutic approach, which consists of preoperative patient information, sufficient analgesia, early mobilisation and enteral feeding, may solve this discrepancy. Therefore, prospective controlled studies are needed to assess the influence of this perioperative approach on outcome. PMID- 9412278 TI - How are ICPs shaping and participating in performance improvement within their respective health care settings, especially through application of surveillance data on nosocomial infections for measuring and improving quality of care? PMID- 9412279 TI - What is the impact of emerging health care delivery systems on ICPs, and what strategies do they use in their respective workplaces to ensure continued success of infection prevention and control programs? PMID- 9412277 TI - ANZICS 21st annual scientific meeting. Melbourne, Victoria, October 17-20, 1996. Abstracts. PMID- 9412280 TI - Emergency medical informatics. PMID- 9412281 TI - Twenty-five years of evolution and revolution: how the specialty has changed. PMID- 9412282 TI - Update on women's health. PMID- 9412283 TI - Ductal carcinoma in situ of the breast. AB - PURPOSE: The increasing incidence and biological heterogeneity of ductal carcinoma in situ (DCIS) of the breast have made the management of this entity challenging and controversial. This paper reviews data on the natural history of the disease and results obtained with various management approaches. DATA SOURCES: Computerized MEDLINE search of articles related to DCIS published since 1966. STUDY SELECTION: Randomized trials were given higher value; however, because these were relatively scarce, retrospective studies and data published in abstract form were also included. DATA EXTRACTION: The authors reviewed all sources critically. No formal statistical calculations were made. DATA SYNTHESIS: The incidence of DCIS is increasing, and a greater proportion of diagnoses are being made in asymptomatic patients. No data from randomized trials compare mastectomy and breast-conserving therapy for the treatment of DCIS. A large randomized trial comparing lumpectomy with lumpectomy plus radiotherapy showed lumpectomy plus radiotherapy to be effective for management of this disease. The presence of comedo necrosis and surgical margin status are frequently used as predictors of subsequent recurrence, although this practice is controversial. The risk for in-breast recurrence at 5 years after lumpectomy and radiotherapy is approximately 8%. With more refined molecular analysis, the relation of DCIS to invasive breast cancer will be better defined. CONCLUSIONS: Treatment strategies for DCIS have evolved, and lumpectomy followed by radiotherapy is an appropriate alternative for most patients. The use of lumpectomy alone in selected patients remains controversial. PMID- 9412285 TI - Lessons from the mammography screening controversy: can we improve the debate? AB - The debate about breast cancer screening for women in their 40s has become so contentious that effective communication and rational discussion on this topic have been compromised. This contentiousness might be defused by understanding the reasons for it. The debate is less about facts than it is about perceptions and values. There is disagreement about how to fairly describe facts about risk and how to avoid misperceptions that may distort assessment of risk. Other sources of disagreement concern the potential harms of screening, the relative roles of physicians and patients in decision making, and how to factor cost into screening decisions. The entire decision-making process has also been highly charged by single-issue advocacy groups and a kind of gender rivalry. Several approaches might help defuse the debate and improve discussion. First, those on both sides of the debate might agree on several things: 1) that the evidence from clinical trials is widely agreed-upon and thus that a main task now is to factor in the values of individual women who are making decisions; 2) that the values of women may differ substantially and that those differences should be respected; 3) that both individuals and the public should be fully and fairly informed about the pros and cons of screening; and 4) that cost-effectiveness should at least be considered during the decision-making process. Lessons from this debate may apply to other medical problems that have small degrees of risk and whose management is strongly debated. PMID- 9412284 TI - Using autopsy series to estimate the disease "reservoir" for ductal carcinoma in situ of the breast: how much more breast cancer can we find? AB - PURPOSE: To determine how many cases of breast cancer might be found if women not known to have the disease were thoroughly examined (the disease "reservoir"). DATA SOURCES: MEDLINE search from 1966 to the present. STUDY SELECTION: Hospital based and forensic autopsy series examining women not known to have had breast cancer during life. DATA EXTRACTION: Observed prevalence of occult invasive breast cancer or ductal carcinoma in situ (DCIS) in which the number of women who were given a diagnosis was the numerator and the number of women examined was the denominator. For each autopsy series, we attempted to ascertain the level of scrutiny (sampling method, number of slides examined) given to the pathologic specimens. DATA SYNTHESIS: Among seven autopsy series of women not known to have had breast cancer during life, the median prevalence of invasive breast cancer was 1.3% (range, 0% to 1.8%) and the median prevalence of DCIS was 8.9% (range, 0% to 14.7%). Prevalences were higher among women likely to have been screened (that is, women 40 to 70 years of age). The mean number of slides examined per breast ranged from 9 to 275; series that reported higher levels of scrutiny tended to discover more cases of cancer. CONCLUSIONS: A substantial reservoir of DCIS is undetected during life. How hard pathologists look for the disease and, perhaps, their threshold for making the diagnosis are potentially important factors in determining how many cases of DCIS are diagnosed. The latter has important implications for what it means to have the disease. PMID- 9412286 TI - Breast cancer screening in women younger than 50 years of age: what's next? PMID- 9412287 TI - Rise and fall. PMID- 9412288 TI - Thromboembolism after cardioversion for atrial fibrillation. PMID- 9412289 TI - Thromboembolism after cardioversion for atrial fibrillation. PMID- 9412290 TI - Osteoporosis and mortality. PMID- 9412291 TI - Risk stratification after myocardial infarction. PMID- 9412292 TI - Molecular diagnosis of thiopurine S-methyltransferase deficiency. PMID- 9412293 TI - Follow-up of sarin poisoning in Matsumoto. PMID- 9412294 TI - Pancreatic fibrosis with islet cell pseudoneoplastic proliferation as a cause of hypoglycemia. PMID- 9412295 TI - Prostacyclin for HIV-associated pulmonary hypertension. PMID- 9412296 TI - Losartan-induced acute pancreatitis. PMID- 9412297 TI - Dichotomous disservice. PMID- 9412298 TI - Dichotomous disservice. PMID- 9412299 TI - The language of medical case histories. PMID- 9412300 TI - Cost-effectiveness of extending screening mammography guidelines to include women 40 to 49 years of age. AB - BACKGROUND: Screening mammography is recommended for women 50 to 69 years of age because of its proven efficacy and reasonable cost-effectiveness. Extending screening recommendations to include women 40 to 49 years of age remains controversial. OBJECTIVE: To compare the cost-effectiveness of screening mammography in women of different age groups. DESIGN: Cost-effectiveness analysis done using Markov and Monte Carlo models. PATIENTS: General population of women 40 years of age and older. INTERVENTIONS: Biennial screening from 50 to 69 years of age was compared with no screening. Screening done every 18 months from ages 40 to 49 years, followed by biennial screening from ages 50 to 69 years, was compared with biennial screening from ages 50 to 69 years. MEASUREMENTS: Life expectancy, costs, and incremental cost-effectiveness. RESULTS: Screening women from 50 to 69 years of age improved life expectancy by 12 days at a cost of $704 per woman, resulting in a cost-effectiveness ratio of $21,400 per year of life saved. Extending screening to include women 40 to 49 years of age improved life expectancy by 2.5 days at a cost of $676 per woman. The incremental cost effectiveness of screening women 40 to 49 years of age was $105,000 per year of life saved. On the basis of a multiway sensitivity analysis, there is a 75% chance that screening mammography in women 50 to 69 years of age costs less than $50,000 per year of life saved, compared with a 7% chance in women 40 to 49 years of age. CONCLUSION: The cost-effectiveness of screening mammography in women 40 to 49 years of age is almost five times that in older women. When breast cancer screening policies are being set, the incremental cost-effectiveness of extending mammographic screening to younger women should be considered. PMID- 9412301 TI - The role of numeracy in understanding the benefit of screening mammography. AB - BACKGROUND: Quantitative information about risks and benefits may be meaningful only to patients who have some facility with basic probability and numerical concepts, a construct called numeracy. OBJECTIVE: To assess the relation between numeracy and the ability to make use of typical risk reduction expressions about the benefit of screening mammography. DESIGN: Randomized, cross-sectional survey. SETTING: A simple random sample of 500 female veterans drawn from a New England registry. INTERVENTION: One of four questionnaires, which differed only in how the same information on average risk reduction with mammography was presented. MEASUREMENTS: Numeracy was scored as the total number of correct responses to three simple tasks. Participants estimated their risk for death from breast cancer with and without mammography. Accuracy was judged as each woman's ability to adjust her perceived risk in accordance with the risk reduction data presented. RESULTS: 61% of eligible women completed the questionnaire. The median age of these women was 68 years (range, 27 to 88 years), and 96% were high school graduates. Both accuracy in applying risk reduction information and numeracy were poor (one third of respondents thought that 1000 flips of a fair coin would result in < 300 heads). Accuracy was strongly related to numeracy: The accuracy rate was 5.8% (95% CI, 0.8% to 10.7%) for a numeracy score of 0, 8.9% (CI, 2.5% to 15.3%) for a score of 1, 23.7% (CI, 13.9% to 33.5%) for a score of 2, and 40% (CI, 25.1% to 54.9%) for a score of 3. CONCLUSIONS: Regardless of how information was presented, numeracy was strongly related to accurately gauging the benefit of mammography. More effective formats are needed to communicate quantitative information about risks and benefits. PMID- 9412302 TI - The role of hormone replacement therapy in the risk for breast cancer and total mortality in women with a family history of breast cancer. AB - BACKGROUND: The risks and benefits of hormone replacement therapy (HRT) are of considerable interest and importance, especially in terms of whether they differ among subsets of women. OBJECTIVE: To determine whether HRT is associated with increased risks for breast cancer and total mortality in women with a family history of breast cancer. DESIGN: Prospective cohort study. SETTING: Population based sample of midwestern post-menopausal women enrolled in an observational study of risk factors for cancer. PARTICIPANTS: Random sample of 41,837 female Iowa residents 55 to 69 years of age. MEASUREMENTS: Incidence rates of and relative risks for breast cancer (n = 1085) and total mortality (n = 2035) through 8 years of follow-up were calculated by using data from the State Health Registry of Iowa and the National Death Index. RESULTS: A family history of breast cancer was reported by 12.2% of the cohort at risk. Among women with a family history of breast cancer, those who currently used HRT and had done so for at least 5 years developed breast cancer at an age-adjusted annual rate of 61 cases per 10,000 person-years (95% CI, 28 to 94 cases); this rate was not statistically significantly higher than the rate in women who had never used HRT (46 cases per 10,000 person-years [CI, 36 to 55 cases]). Among women with a family history, those who used HRT had a significantly lower risk for total mortality than did women who had never used HRT (relative risk, 0.67 [CI, 0.51 to 0.89]), including total cancer-related mortality (relative risk, 0.75 [CI, 0.50 to 1.12]). The age-adjusted annual mortality rate for women using HRT for at least 5 years was 46 deaths per 10,000 person-years (CI, 19 to 74 deaths); this is roughly half the rate seen in women who had never used HRT (80 deaths per 10,000 person-years [CI, 69 to 92 deaths]). CONCLUSIONS: These data suggest that HRT use in women with a family history of breast cancer is not associated with a significantly increased incidence of breast cancer but is associated with a significantly reduced total mortality rate. PMID- 9412303 TI - Reversibility of hepatic fibrosis in autoimmune hepatitis. AB - BACKGROUND: Hepatic fibrosis and cirrhosis occur in many types of chronic liver injury and generally seem to be irreversible. OBJECTIVE: To determine whether cirrhosis caused by autoimmune hepatitis can be reversible. DESIGN: Retrospective study. PATIENTS: Eight patients with autoimmune hepatitis and cirrhosis who responded to medical therapy and had follow-up liver biopsy while in clinical and biochemical remission. MEASUREMENTS: Biopsy specimens were randomly coded in an unpaired manner according to patient and were read independently by two pathologists using the Knodell scoring system. RESULTS: The median alanine aminotransferase level decreased from 10.30 mukat/L to 0.37 mukat/L, the median serum bilirubin level decreased from 70 mumol/L to 10 mumol/L, and the median serum albumin level increased from 34 g/L to 43 g/L. Cirrhosis, extensive fibrosis, or both were present in all patients at diagnosis but were not present on follow-up liver biopsy. The median Knodell score decreased from 14.0 to 1.3, and the median fibrosis score decreased from 3.3 to 0.8. CONCLUSION: Hepatic fibrosis and cirrhosis may be reversible in some patients in whom autoimmune hepatitis responds to treatment. PMID- 9412304 TI - Effect of fluoroquinolone on the enhanced nitric oxide-induced peripheral vasodilation seen in cirrhosis. AB - BACKGROUND: In patients with cirrhosis, portosystemic shunts allow intestinal bacteria and endotoxin to enter the systemic circulation. Endotoxemia may induce increased synthesis of nitric oxide, thereby contributing to arterial vasodilation. OBJECTIVE: To test the hypothesis that the antibiotic norfloxacin blocks the effects of nitric oxide. DESIGN: Placebo-controlled, double-blind, crossover study. SETTING: Alfred Hospital, Melbourne, Australia. PATIENTS: 9 patients with alcohol-related cirrhosis and 10 healthy controls. INTERVENTION: Norfloxacin, 400 mg twice daily, for 4 weeks. MEASUREMENTS: Peripheral blood flow was measured by using forearm venous occlusion plethysmography. RESULTS: Basal forearm blood flow was higher in patients with cirrhosis than in controls (3.69 +/- 0.27 mL/100 mL per minute and 2.47 +/- 0.40 mL/100 mL per minute; P = 0.014) but returned toward normal after norfloxacin was given (2.64 +/- 0.31 mL/100 mL of tissue per minute in patients with cirrhosis). Responses to NG-monomethyl-L arginine were greater in patients with cirrhosis but returned to normal after norfloxacin was given. CONCLUSION: Bacterial endotoxemia in patients with cirrhosis induces increased synthesis of nitric oxide that can be corrected with norfloxacin. PMID- 9412305 TI - Integrating heterogeneous pieces of evidence in systematic reviews. AB - Researchers preparing systematic reviews often encounter various types of evidence, which can generally be categorized as direct or indirect. The former directly relates an exposure, diagnostic strategy, or therapeutic intervention to the occurrence of a principal health outcome. Evidence is indirect if two or more bodies of evidence are required to relate the exposure, diagnostic strategy, or intervention to the principal health outcome. Heterogeneity of data sources complicates integration of both direct and indirect evidence. Participants in different studies may have a wide spectrum of baseline risk and sociodemographic and cultural characteristics. A variety of formulations and intensities of exposures, diagnostic strategies, and interventions, as well as diversity in the selection and definition of control groups, may be encountered. Outcome measures may be different, and similar outcomes may be measured or reported differently. Heterogeneity of study designs and of methodologic features and quality within a given design may be found. The effective integration of direct and indirect evidence requires development of explicit models that serve as analytic frameworks for linking the important pieces of evidence. A model can be viewed as a series of subquestions, with each important subquestion warranting a systematic review. Several subjective and quantitative methods can then be used to integrate the evidence. Tabular displays of major findings and strength of evidence for each subquestion can help reviewers, patients, and providers to integrate the differing research findings and draw reasonable conclusions. Various quantitative techniques, such as decision analysis and the confidence profile method, are also available. No single integration approach is clearly superior, none obviates uncertainty, and all underscore the role of careful judgment in integrating evidence. PMID- 9412306 TI - A critical pathway for management of patients with acute chest pain who are at low risk for myocardial ischemia: recommendations and potential impact. AB - BACKGROUND: Use of resources for patients with acute chest pain may be improved with clinical strategies that integrate research, Bayesian analysis, and expert opinion. OBJECTIVES: To 1) develop a critical pathway for management of patients with acute chest pain who are at low risk for complications of ischemic heart disease and 2) assess the potential effects of implementation of the pathway on patient safety and resource use. DESIGN: Evidence-based consensus and prospective cohort study. SETTING: Urban teaching hospital. PATIENTS: Patients at least 30 years of age who were seen in the emergency department for chest pain and who did not have a history of trauma or abnormalities on radiologic study. INTERVENTION: Physician-opinion leaders defined criteria for patient inclusion in the pathway and for remaining on the pathway after 6 or 12 hours of observation. Criteria were defined for appropriateness of direct admission, direct discharge, or 6 hours of observation followed by exercise treadmill testing. MEASUREMENTS: Number of patients admitted to the hospital, number of days that patients were hospitalized, and clinical outcome. RESULTS: 2898 of 4585 patients (63%) were admitted to the hospital; of the 2898, 1152 (40%) were classified as potentially eligible for the pathway and 1068 (93%) had a benign clinical course during the initial observation period. The 1068 patients had a mean length of stay of 2.8 +/ 4.8 days. If 47% of these patients had been discharged after observation and exercise testing, implementation of the pathway would have reduced the number of admissions by 505 (17%) and days of hospitalization by 1407 (11%). CONCLUSIONS: Retrospective analysis suggests that a critical pathway for patients with acute chest pain may substantially reduce resource use. Prospective study is needed to ensure increased efficiency without increased adverse outcomes. PMID- 9412307 TI - Monitored isoniazid prophylaxis for low-risk tuberculin reactors older than 35 years of age: a risk-benefit and cost-effectiveness analysis. AB - BACKGROUND: Isoniazid chemoprophylaxis effectively prevents the development of active infectious tuberculosis. Current guidelines recommend withholding this prophylaxis for low-risk tuberculin reactors older than 35 years of age because of the risk for fatal isoniazid-induced hepatitis. However, recent studies have shown that monitoring for hepatotoxicity can significantly reduce the risk for isoniazid-related death. OBJECTIVE: To evaluate the effectiveness and cost effectiveness of monitored isoniazid prophylaxis for low-risk tuberculin reactors older than 35 years of age. DESIGN: A Markov model was used to compare the health and economic outcomes of prescribing or withholding a course of prophylaxis for low-risk reactors 35, 50, or 70 years of age. Subsequent analyses evaluated costs and benefits when the effect of transmission of Mycobacterium tuberculosis to contacts was included. MEASUREMENTS: Probability of survival at 1 year, number needed to treat, life expectancy, and cost per year of life gained for individual persons and total population. RESULTS: Isoniazid prophylaxis increased the probability of survival at 1 year and for all subsequent years. For 35-year old, 50-year-old, and 70-year-old tuberculin reactors, life expectancy increased by 4.9 days, 4.7 days, and 3.1 days, respectively, and costs per person decreased by $101, $69, and $11, respectively. When the effect of secondary transmission to contacts was included, the gains in life expectancy per person receiving prophylaxis were 10.0 days for 35-year-old reactors, 9.0 days for 50-year-old reactors, and 6.0 days for 70-year-old reactors. Costs per person for these cohorts decreased by $259, $203, and $100, respectively. The magnitude of the benefit of isoniazid prophylaxis is moderately sensitive to the effect of isoniazid on quality of life. The hypothetical provision of isoniazid prophylaxis for all low-risk reactors older than 35 years of age in the U.S. population could prevent 35,176 deaths and save $2.11 billion. CONCLUSIONS: Monitored isoniazid prophylaxis reduces mortality rates and health care costs for low-risk tuberculin reactors older than 35 years of age, although reductions for individual patients are small. For the U.S. population, however, the potential health benefits and economic savings resulting from wider use of monitored isoniazid prophylaxis are substantial. We should consider expanding current recommendations to include prophylaxis for tuberculin reactors of all ages with no contraindications. PMID- 9412308 TI - Somatostatin or octreotide compared with H2 antagonists and placebo in the management of acute nonvariceal upper gastrointestinal hemorrhage: a meta analysis. AB - PURPOSE: To determine the efficacy of somatostatin or octreotide for the treatment of acute nonvariceal upper gastrointestinal hemorrhage. DATA SOURCE: Database searches of English-language articles published between 1966 and 1996 and the bibliographies of all related articles and textbook chapters. STUDY SELECTION: Randomized clinical trials comparing somatostatin or octreotide with H2 blockers or placebo in patients with a clinical or endoscopic diagnosis of acute nonvariceal upper gastrointestinal hemorrhage. DATA EXTRACTION: Methods and quality of the studies were evaluated, and quantitative data on outcomes, including continued bleeding, rebleeding during the treatment period, need for surgery, and transfusion requirement, were extracted. DATA SYNTHESIS: Among 1829 patients from 14 trials, the relative risk (RR) for continued bleeding or rebleeding was 0.53 (95% CI, 0.43 to 0.63) in favor or somatostatin, with a number needed to treat (NNT) of 5. Among 7 investigator-blinded trials, the relative risk was 0.73 (CI, 0.64 to 0.81) and the NNT was 11. Somatostatin was efficacious for peptic ulcer bleeding (RR, 0.48 [CI, 0.39 to 0.59]; NNT, 4) and showed a trend toward efficacy for non-peptic ulcer bleeding (RR, 0.62 [CI, 0.39 to 1.002]). Although the overall results suggested a decreased need for surgery in the somatostatin group, a subgroup analysis of investigator-blinded trials revealed a more modest effect that was not statistically significant (RR, 0.94 [CI, 0.87 to 1.001]). CONCLUSION: Somatostatin may reduce the risk for continued bleeding from acutely bleeding peptic ulcer disease. Somatostatin may be useful either as an adjunct treatment before endoscopy or when endoscopy is unsuccessful, contraindicated, or unavailable. PMID- 9412309 TI - Chronic obstructive pulmonary disease stage and health-related quality of life. The Quality of Life of Chronic Obstructive Pulmonary Disease Study Group. AB - BACKGROUND: The American Thoracic Society recently recommended that chronic obstructive pulmonary disease be staged on the basis of the percentage of predicted FEV1. OBJECTIVE: To examine 1) the relation between the american Thoracic Society system for staging chronic obstructive pulmonary disease and health-related quality of life and 2) the effect of self-reported comorbid conditions on health-related quality of life. DESIGN: Cross-sectional study. SETTING: Outpatient clinics of respiratory departments of four hospitals and one primary health care center in spain. PATIENTS: 321 consecutive male patients with chronic obstructive pulmonary disease. MEASUREMENTS: Functional respiratory impairment, FEV1, respiratory symptoms, and health-related quality of life. Respiratory symptoms and health-related quality of life were measured by using the Spanish version of the St. George's Respiratory Questionnaire and the Nottingham Health Profile. RESULTS: Patient scores on the St. George's Respiratory Questionnaire were moderately to strongly associated with disease staging (r = 0.27 to 0.51). Compared with reference values, values for health related quality of life for patients with stage I disease were substantially higher on the St. George's Respiratory Questionnaire (6 and 34; p < 0.001) and values for impairment were significantly greater in stage 1 patients with comorbid conditions (19 and 36; P = 0.001). At least one concomitant chronic condition was found in 84% of study patients. Comorbid conditions only partly influenced the observed pattern of deterioration of health-related quality of life with worsening stages of disease. CONCLUSION: Staging criteria for chronic obstructive pulmonary disease based on percentage of predicted FEV1 separated groups of patients with varying degrees of impairment in health-related quality of life. Contrary to expectations, even patients with mild disease showed substantially compromised health-related quality of life. Comorbid conditions influenced the relation between chronic obstructive pulmonary disease and health related quality of life. PMID- 9412311 TI - Treatment of hyperhomocysteinemia in renal transplant recipients. A randomized, placebo-controlled trial. AB - BACKGROUND: Stable renal transplant recipients have an excess prevalence of hyperhomocysteinemia, which is a risk factor for arteriosclerosis. OBJECTIVE: To determine the effect of treatment with 1) vitamin B6 or 2) folic acid plus vitamin B12 on fasting and post-methionine-loading plasma total homocysteine levels in renal transplant recipients. DESIGN: Block-randomized, placebo controlled, 2 x 2 factorial study. SETTING: University-affiliated transplantation program. PATIENTS: 29 clinically stable renal transplant recipients. INTERVENTION: Patients were randomly assigned to one of four regimens: placebo (n = 8); vitamin B6, 50 mg/d (n = 7); folic acid, 5 mg/d, and vitamin B12, 0.4 mg/d (n = 7); or vitamin B6, 50 mg/d, folic acid, 5 mg/d, and vitamin B12, 0.4 mg/d (n = 7). MEASUREMENTS: Fasting and 2-hour post-methionine-loading plasma total homocysteine levels. RESULTS: Vitamin B6 treatment resulted in a 22.1% reduction in geometric-mean post-methionine-loading increases in plasma total homocysteine levels (P = 0.042), and folic acid plus vitamin B12 treatment caused a 26.2% reduction in geometric-mean fasting plasma total homocysteine levels (P = 0.027). These results occurred after adjustment for age; sex; and pretreatment levels of total homocysteine, B vitamins, and creatinine. CONCLUSIONS: Vitamin B6 should be added to the combination of folic acid and vitamin B12 for effective reduction of both post-methionine-loading and fasting plasma total homocysteine levels in renal transplant recipients. PMID- 9412310 TI - Unrelated donor bone marrow transplantation for chronic myelogenous leukemia: a decision analysis. AB - BACKGROUND: Chronic myelogenous leukemia (CML) is an indolent but ultimately fatal disease. Because the natural history of CML varies and quality of life with CML may be excellent until shortly before death, deciding whether and when to pursue unrelated donor bone marrow transplantation is often difficult. OBJECTIVE: To compare early transplantation, delayed transplantation, and no transplantation for patients with chronic-phase CML on the basis of discounted, quality-adjusted life expectancy. DESIGN: A markov model comparing different strategies was constructed. This model considers patient age, quality of life, risk aversion, and the competing risks for CML progression and transplant toxicity. SETTING: Therapeutic decision at the time of diagnosis of CML. PATIENTS: The base case is a 35-year-old patient with intermediate-prognosis CML. Younger and older patients with better and worse prognoses are also evaluated. INTERVENTION: Early transplantation, delayed transplantation, and no transplantation. MEASUREMENTS: Quality-adjusted, discounted life expectancy. RESULTS: For patients with newly diagnosed CML, transplantation within the first year provides the greatest quality-adjusted expected survival, although this benefit decreases with increasing patient age. For a 35-year-old patient with intermediate-prognosis CML, transplantation within the first year results in 53 more discounted, quality adjusted years of life expectancy than does no transplantation. This finding is robust even with varying baseline assumptions. CONCLUSIONS: These results support the use of early unrelated donor bone marrow transplantation for most patients with CML. PMID- 9412312 TI - Personal use of postmenopausal hormone replacement therapy by women physicians in the United States. AB - BACKGROUND: Women physicians' use of postmenopausal hormone replacement therapy (HRT) is unknown. OBJECTIVE: To study use of HRT by women physicians in the United States. DESIGN: Stratified random-sample mail survey. SETTING: United States Participants: 1466 postmenopausal women U.S. physicians in the Women Physicians' health study. MEASUREMENTS: Self-reported personal use of HRT and information on demographic, professional, and behavioral characteristics and medical history. RESULTS: Overall, 47.4% of participants currently use HRT; the prevalence of use is 59.8% in women 40 to 49 years of age, 49.4% in women 50 to 59 years of age, and 36.4% in women 60 to 70 years of age (P < 0.001). In an adjusted logistic regression model, current users were significantly more likely to be gynecologists, to be younger, to be white, to be sexually active, to be previous users of oral contraceptives, to live in Pacific or Mountain states, to have had a hysterectomy, and to have no personal or family history of breast cancer. CONCLUSIONS: Women physicians have a higher rate of HRT use than that reported in cross-sectional U.S. surveys. This may presage greater use of HRT for U.S. women in the future. PMID- 9412313 TI - Collaborative management of chronic illness. AB - In chronic illness, day-to-day care responsibilities fall most heavily on patients and their families. Effective collaborative relationships with health care providers can help patients and families better handle self-care tasks. Collaborative management is care that strengthens and supports self-care in chronic illness while assuring that effective medical, preventive, and health maintenance interventions take place. In this paper, the following essential elements of collaborative management developed in light of behavioral principles and empirical evidence about effective care in chronic illness are discussed: 1) collaborative definition of problems, in which patient-defined problems are identified along with medical problems diagnosed by physicians; 2) targeting, goal setting, and planning, in which patients and providers focus on a specific problem, set realistic objectives, and develop an action plan for attaining those objectives in the context of patient preferences and readiness; 3) creation of a continuum of self-management training and support services, in which patients have access to services that teach skills needed to carry out medical regimens, guide health behavior changes, and provide emotional support; and 4) active and sustained follow-up, in which patients are contacted at specified intervals to monitor health status, identify potential complications, and check and reinforce progress in implementing the care plan. These elements make up a common core of services for chronic illness care that need not be reinvented for each disease. PMID- 9412314 TI - The changing clinical spectrum of adrenal insufficiency. PMID- 9412315 TI - Guidelines for laboratory evaluation in the diagnosis of Lyme disease. American College of Physicians. PMID- 9412316 TI - Laboratory evaluation in the diagnosis of Lyme disease. AB - PURPOSE: To provide a qualitative evaluation of the predictive value of the laboratory diagnosis of Lyme disease and to use the resultant data to formulate guidelines for clinical diagnosis. DATA SOURCES: A MEDLINE search of English language articles or articles with English-language abstracts published from 1982 to 1996. DATA EXTRACTION: Sensitivity, specificity, and likelihood ratios were calculated, and a random-effects model was used to combine the proportions from the eligible studies. Prespecified criteria were used to determine which studies were eligible for analysis. DATA SYNTHESIS: Laboratory testing in general is not clinically useful if the pretest probability of Lyme disease is less than 0.20 or greater than 0.80. When the pretest probability is 0.20 to 0.80, sequential testing with enzyme-linked immunosorbent assay and Western blot is the most accurate method for ruling in or ruling out the possibility of Lyme disease. CONCLUSIONS: Laboratory testing is recommended only in patients whose pretest probability of Lyme disease is 0.20 to 0.80. If the pretest probability is less than 0.20, testing will result in more false-positive results than true-positive results; a negative test result in this situation effectively rules out the disease. PMID- 9412317 TI - Chronic active adolescence. PMID- 9412318 TI - Hypercoagulable states. PMID- 9412319 TI - Hypercoagulable states. PMID- 9412320 TI - Risk factors for deep venous thrombosis of the upper extremities. PMID- 9412321 TI - Meningitis and skin reaction after intravenous immune globulin therapy. PMID- 9412322 TI - Meningitis and skin reaction after intravenous immune globulin therapy. PMID- 9412323 TI - Weekly fluconazole for preventing mucosal candidiasis in HIV infection. PMID- 9412324 TI - Depression, smoking, and health status. PMID- 9412325 TI - Rethinking somatization. PMID- 9412326 TI - Rethinking somatization. PMID- 9412327 TI - Rethinking somatization. PMID- 9412328 TI - Rethinking somatization. PMID- 9412329 TI - Medicinal uses of marijuana. PMID- 9412330 TI - Medicinal uses of marijuana. PMID- 9412331 TI - Medicinal uses of marijuana. PMID- 9412332 TI - Intractable hiccups and gastroesophageal reflux disease. PMID- 9412334 TI - Long, benign course of hepatitis G virus infection. PMID- 9412335 TI - Lemon-yellow nails and long-term phenazopyridine use. PMID- 9412333 TI - Recurrent severe acute hepatitis and omeprazole. PMID- 9412336 TI - The man with stars inside. PMID- 9412337 TI - The man with stars inside. PMID- 9412338 TI - [A severe form of urinary tuberculosis in children]. AB - Urinary tuberculosis is a rare disease in children. It poses major diagnostic problems because of clinical symptoms, which are often atypical and misleading. It causes serious lesions which are often multifocal and extensive, requiring complex surgical excision and urinary tract reconstruction. Prevention of this disease is based on generalized vaccination with BCG and adequate treatment of pulmonary tuberculosis. The authors report a case of urinary tuberculosis in a fourteen-year-old child who presented episodes of cystitis and hematuria refractory to treatment. The diagnosis, confirmed by the positive test for AFB in the urine was established late, at the stage of silent kidney and scleroatrophic bladder. The patient was treated with antituberculous chemotherapy (Isoniazid; Rifampicin, PZA) and nephro-ureterectomy with augmentation enterocystoplasty. PMID- 9412339 TI - [An unusual cause of renal colic. An osteophytic spur of L3, surgically treated with success]. AB - The authors report an unusual case of renal colic occurring in a 44-year-old docker. Intravenous urography showed right ureteral extrinsic compression by an osteophyte of the 3rd lumbar vertebra. After failure of medical treatment, the patient was operated with resection of the osteophyte. The postoperative course was uneventful with reduction of pain. PMID- 9412340 TI - [Congenital cysts of the seminal vesicles with ipsilateral kidney agenesis. Clinical aspects of 7 cases]. AB - Seminal vesicle cysts with ureteral ectopy and ipsilateral renal agenesis or dysplasia are rare congenital malformations. This study reports 7 consecutive cases in a single center and, for the first time in the literature, reports long term results on 5 cases. Patients were between the age of 15 and 57 years. The malformation was diagnosed in all cases with excretory urography, computed tomography, magnetic resonance imaging and cystoscopy. 5 patients were followed after diagnosis for 26 to 119 months (mean 52 months). Only 2 of 7 patients primarily presented with nonspecific lower urinary tract symptoms. One patient (15 years old) showed significant enlargement of the cysts with compression of the urinary bladder and signs of urinary incontinence 10 years after primary diagnosis. The other 4 patients had no changes of their malformation. In conclusion, seminal vesicle cysts with ipsilateral renal agenesis are now more frequently diagnosed because of the increasing use of sectional imaging procedures. They are mostly asymptomatic and their morphology does not change with time. These data support the concept of only treating symptomatic patients. PMID- 9412342 TI - [Cystitis cystica glandularis. A study of 2 cases]. AB - The authors report two cases of cystitis glandularis revealed by hematuria and urinary retention. Cystitis glandularis is a rare disease, caused by metaplasia of the vesical submucosa, probably related to a chronic irritative factor. In its minor form, it has the same clinical features as simple cystitis, but its major pseudoneoplastic form may be mistaken for bladder tumor. The diagnosis is essentially histological. Treatment is usually medical, based on eradication of the irritative factors. Surgery is performed in the case of complications of this disease. The clinical course is unclear, requiring long-term surveillance. PMID- 9412343 TI - [Epidermoid carcinoma of the bladder. Apropos of 14 cases]. AB - The authors report 14 cases of squamous cell carcinoma of bladder over a period of 20 years. All patients were male, with a mean age of 56 years. A history of urogenital bilharziasis was reported in 2 patients and 10 patients were smokers. Hematuric cystitis was the commonest presenting feature and infiltration of the bladder base was detected on rectal palpation in 10 patients. The patients consulted late in the course, at the stage of upper urinary tract repercussions (11 cases) and computerized tomography showed locoregional extension in 5 cases. 12 total cystoprostatectomies were performed and 2 patients were treated exclusively by radiotherapy. The long-term course was marked by urethral recurrence in 3 patients. The etiopathogenesis, diagnosis, treatment and clinical course of these tumors, which are often aggressive, are reviewed. PMID- 9412344 TI - [Esophageal metastasis of cancer of the bladder]. AB - The authors report a case of oesophageal metastasis of transitional cell carcinoma of the bladder. This lesion was responsible for hematemesis eight days after cystoprostatectomy. Treatment consisted of irradiation of the metastatic site and chemotherapy. The patient is in complete remission after a follow-up of 2 years. PMID- 9412341 TI - [Calculus-induced anuria. Apropos of 25 cases]. AB - The objective of this study was to analyse the clinical and therapeutic aspects of stone-induced anuria based on the analysis of 25 cases. The mean age of the patients was 51 years. The stones were bilateral in 54% of cases, and consisted of uric acid stones in 52% of cases. The diagnosis was based on laboratory and radiological findings. Dialysis was indicated in eight patients because of major metabolic disorders. Urinary diversion was performed by insertion of a ureteric catheter or percutaneous nephrostomy. The medium- to long-term prognosis was favourable in two-thirds of cases, but 3 immediate deaths were observed due to delayed diagnosis. PMID- 9412345 TI - [An immediate measurement of intravesical pressure. A natural method of a urodynamic test in children with myelodysplasia]. AB - A comparison of bladder leak point pressure values measured during conventional filling cystometry with 8 F, and then with 4-5 F catheter was performed in 20 infants with myelodysplasia. In 12 (60%) of them, the larger catheter led to artificial findings, especially in children less than one year old. This was observed in 9 (82%) of 11 infants in this age-group. The method of impromptu bladder pressure monitoring has been introduced, in order to eliminate the risk of artificial results. It is based on testing an already spontaneously filled bladder with a fine catheter without any previous instrumentation and residual bladder emptying. The rationale of this method of urodynamic testing, based on the results of 36 investigated infants, is discussed. There is no need for special equipment, standard examination conditions close to natural conditions and time saving constitute the main advantages of this method. It can be proposed as a first step of prognostic urodynamic testing especially in the early postnatal period and in children less than two years old. PMID- 9412346 TI - [Recurrence of cancer of the prostate after initial treatment with diethylstilbestrol (DES) in a homogeneous series of 175 cases]. AB - The study of a homogeneous series of 215 cases of prostatic cancer, 175 of which received first-line treatment with diethylstilbestrol (DES) clearly demonstrated that: 1. practically all well differentiated prostatic cancers, regardless of their stage, responded remarkably well to first-line treatment with sufficient doses of DES, as, when correctly monitored, practically none of these cancers escaped and early stages of escape can be salvaged. 2. true escape occurs all the earlier and evolves all the more rapidly for advanced, poorly differentiated cancers, but this is not constant. 3. to our knowledge, confirmed true metastatic escape to effective doses of DES cannot be salvaged by another treatment, while DES has been successfully used as salvage therapy in several cases of escape to LHRH agonists. 4. the thromboembolic risks must be thoroughly evaluated and prevented by associated anticoagulant treatment. They do not constitute a formal contraindication to treatment when it is closely monitored. However, the best treatment for these cases, when accepted by the patient, appears to be a combination of castration with a daily dose of 1 to 2 mg of DES. 5. The low cost of this treatment must also be taken into account in the assessment of this treatment modality. PMID- 9412347 TI - [Polyps of the posterior urethra in children. Apropos of a case]. AB - Posterior urethral polyp is a rare disease, predominantly affecting boys. The authors report the case of an 10-month-old boy admitted to hospital with acute urine retention. The diagnosis of polyp was established on cystourethrography, ultrasonography and endoscopy. The polyp was resected via a bladder incision after failure of transurethral resection. Posterior urethral polyp is a benign tumour whose clinical presentation is dominated by signs of lower urinary tract obstruction. Cystourethrography and ultrasonography confirm the diagnosis and assess the repercussions on kidneys and bladder. Endoscopy has a double role: diagnostic and therapeutic. PMID- 9412348 TI - How to transform a perioperative nursing practice idea into an invention. PMID- 9412349 TI - Perioperative nurse turned inventor. PMID- 9412350 TI - Holistic practice is a tradition of excellence in perioperative nursing. PMID- 9412351 TI - Pew Commission's State Initiatives Program to Reform Health Care Workforce Regulation grant activity update. PMID- 9412352 TI - Selecting research instruments to measure the reliability and validity of nursing research studies. PMID- 9412353 TI - [Urodynamics, neurourology, and neuroandrology]. PMID- 9412354 TI - [Value of the shape of the flowmetry curve in the study of prostatism]. AB - OBJECTIVE: To determine the relation of the urinary flow curve morphology of free flowmetry with the status of lower urinary tract dynamics. METHODS: A mathematical model was designed to fit the urinary flow curves to two models: a symmetrical and an asymmetrical model. Based on these models we analyzed the relationship between the presence or absence of obstruction and the type of model which better adjusted the urinary flow curves in a series of 85 males. RESULTS: The urinary flow curves corresponding to absence of obstruction adjusted significantly better to a symmetrical model than those corresponding to bladder outlet obstruction. No correlation was observed between the type of curve and bladder contractibility or type of urinary obstruction. CONCLUSIONS: There is a relationship between the form of the urinary flow curve and bladder obstruction. In the absence of obstruction, the urinary flow curves are more symmetrical. PMID- 9412355 TI - [Fluid loss pressure in 45 children with congenital neurogenic bladder]. AB - OBJECTIVE: This study analyzed the relation between leak point pressure (LPP) at first urodynamic evaluation and the status of the upper urinary tract (UUT) and renal reflux (RR). METHODS: The study comprised 45 myelodysplastic children, one week to 13 years of age; 19 (42%) were less than one year old. LPP was measured when liquid started to come out through the urethral meatus, around the 5-6 Fr catheter. UUT was evaluated by ultrasound and RR by voiding cystography. RESULTS: 19 children had LPP < 30 cm H2O; all cases had a normal UUT, although 5 (26%) had RR. Twenty-six cases (58%) had LPP > 30 cm H2O; 12 (46%) had abnormal UUT and 6 of these had RR; 7 cases had RR but normal UUT. The group with LPP > or = 30 cm H2O was analyzed according to LPP values. The UUT was abnormal in 31% of cases with LPP 30-60 cm H2O and 37% had RR; UUT was abnormal in 70% of cases with LPP > 60 cm H2O and 70% had RR. Of the 19 patients less than one year old 9 (47%) had LPP < 30 cm H2O, no patient had abnormal UUT and only two (22%) had RR; 10 cases had LPP > or = 30 cm H2O, 6 of these had abnormal UUT and 5 had RR. CONCLUSIONS: In this study UUT was normal when LPP was < 30 cm H2O. Renal impairment and RR increased with LPP. Urethral functional obstruction carries a worse prognosis of pediatric neurogenic bladder. In these cases clean intermittent catheterization is recommended, regardless of patient age and sex. PMID- 9412356 TI - [Correlation coefficient of urodynamic data used in prostatism]. AB - OBJECTIVE: To determine the usefulness of different urodynamic data and their relationship. METHODS: 105 adult males with prostatism were evaluated urodynamically. Data were obtained utilizing the Abrhams and Griffiths and the Schaeffer models. RESULTS: Our results show that the urinary flow rate correlates directly with bladder contractibility and inversely with the urethral resistance parameters. CONCLUSIONS: The urethral resistance was produced by two independent factors: a factor which resists initiation of voiding, measured by the opening pressure according to the Schaeffer model; a second factor which opposes maintenance of voiding, measured by the PURR curvature of the Schaeffer model and by the slope of the Abrhams and Griffiths model. Bladder contractibility was related to pressure at maximum flow rate of the Schaeffer model. PMID- 9412357 TI - [Effect of aging on detrusor function in males]. AB - OBJECTIVE: The present study analyzed the effects of aging on detrusor function in the male. METHODS: The study comprised 200 male patients with prostatism that had undergone complete urodynamic evaluation. The relation of patient age and the clinical and urodynamic data were analyzed. RESULTS: Aging correlates with an increased urinary frequency and a reduced urinary volume and maximum flow rate. Bladder instability was significantly considerably more frequent in those over 80 years of age and the detrusor gradually manages lower maximum voiding pressures with increasing age. On the other hand, urodynamically diagnosed lower urinary tract obstruction and compromise of detrusor contractility are not age-dependent. CONCLUSION: Aging is associated with changes in lower urinary tract function, such as increased urinary frequency, reduced urinary volume, bladder instability is more frequent, reduced maximum voiding pressures, but does not impair detrusor contractility. PMID- 9412358 TI - [Relationship between female stress urinary incontinence intensity and the data of urethral pressure profile]. AB - OBJECTIVE: To determine the utility of the urethral pressure profile in the diagnosis of stress urinary incontinence and its possible correlation with the degree of severity of incontinence. METHODS: 175 female patients with a clinical history of urinary incontinence were evaluated; of these, 50 cases with bladder instability demonstrated by the urodynamic studies were excluded. Patient evaluation included clinical history, physical examination, analytical studies, radiological evaluation and complete urodynamic assessment, including uroflowmetry, filling and voiding cystometry, and static and dynamic urethral pressure profiles. A 10 Fr microtransducer catheter was utilized for the urethral pressure profile studies. ICS recommendations were observed. Patients were classified into three groups according to the severity of urinary incontinence based on the clinical data, physical examination and urodynamic findings. The Wilcoxson test and 2 x 2 contingency table were employed for the statistical analysis. RESULTS: Of the parameters analyzed for the static urethral pressure profile, statistically significant differences were found only for the maximum urethral pressure and maximum closing urethral pressure in the different groups of patients. No differences in total length or functional urethral length were observed. Comparison of the dynamic urethral pressure profiles of the different groups showed a statistically significantly higher proportion of patients with a negative dynamic urethral closing pressure in the group of patients with urodynamically and clinically demonstrated urinary incontinence than in those with no urodynamically or clinically demonstrable incontinence. CONCLUSIONS: The urethral pressure profile is sufficiently reliable to confirm the diagnosis of urinary incontinence and its degree of severity. As a diagnostic test in urinary stress incontinence, it has a sensitivity of 89% and a specificity of 95%. PMID- 9412359 TI - [Artificial urinary sphincter in the treatment of urinary incontinence caused by sphincter insufficiency]. AB - OBJECTIVE: To evaluate the results relative to continence and the complications of the artificial urinary sphincter in the treatment of urinary incontinence due to sphincter incompetence. METHODS: The artificial urinary sphincter model AS 800 was implanted in 35 patients with incontinence arising from various causes. Follow-up ranged from 14 to 108 months (mean 39.8). Patients ages ranged from 4 to 68 years (mean 44.4). There were 17 female (49%) and 18 male patients (51%). The cuff was implanted around the bladder neck in 25 patients (71%) and around the bulbous urethra in 10 (29%). RESULTS: There were 2 mechanical device failures, 2 pump erosion and 1 tube erosion through the abdominal wall in a patient who had had a perforation of an augmentation cystoplasty. Thirty-one patients (88%) became continent. Ten patients (28%) had another operation for device revision or relapse. CONCLUSION: The artificial urinary sphincter is a valid alternative in the treatment of urinary incontinence due to sphincteric incompetence. It is possible to achieve good continence rates, although there is a high revision rate and lifetime follow-up is required. PMID- 9412360 TI - [Psychogenic urinary retention. Diagnostic-therapeutic approach]. AB - OBJECTIVE: The influence of psychogenic factors on voiding generally manifests as an irritative syndrome and rarely in the form of acute or chronic urinary retention. The diagnosis and treatment of this uncommon urological pathology are reviewed and our experience is presented. METHODS: We conducted a retrospective study on 5 patients with psychogenic urinary retention (3 males and 2 females), aged 20 to 28 years (mean age 23.4), that had been treated at our urological services over the last 6 years. Three patients (2 males and 1 female) had a history of depression, one patient had a somatic form of disorder (mimicking) and one patient was diagnosed as having schizophrenia one year after he had presented with urinary retention. The physical and neurological examinations were normal in all 5 patients and the radiological evaluation was normal in all but one patient who had bilateral hydronephrosis. The pressure/flow test disclosed absence of detrusor muscle contraction in all 5 patients; 3 had incomplete voiding by abdominal pressure and had more than 500 ml residual urine. All patients received psychiatric therapy, and intermittent catheterization and urinary rehabilitation until residual urine less than 100 ml was achieved. CONCLUSIONS: The importance of the urodynamic study in the diagnosis of this condition is underscored. Definitive diagnosis can only be established after discarding other pathologies. The initial treatment must always be conservative; irreversible surgical procedures must not be performed. Treatment is by intermittent catheterization, urinary rehabilitation and supportive psychiatric therapy. PMID- 9412361 TI - [Urodynamics of enterocystoplasty and continent diversions]. AB - OBJECTIVE: To describe the filling and voiding urinary dynamics of enterocystoplasty and continent urinary diversion. METHODS: The different behaviour of the tubularized and detubularized intestinal segments and the different sphincteric lesions related with various surgical techniques are described. The possible causes of incontinence are discussed and the literature briefly reviewed. RESULTS/CONCLUSIONS: We underscore the importance of detubularizing the intestinal segment and preserving the sphincteric system as far as possible. Furthermore, voiding using abdominal pressure may cause post void residual urine, dilatation of the neobladder and renal changes. If warranted, self-catheterization should be performed. PMID- 9412362 TI - [Urodynamics in kidney transplantation]. AB - OBJECTIVES: The importance of establishing a protocol for the urological evaluation of patients awaiting renal transplantation is due to the fact that 30% of these adult patients and up to 70% of these children develop renal failure due to a urological condition. The present study analyzes the importance of urodynamic evaluation in these patients. METHODS: 650 patients awaiting renal transplantation were evaluated by patient history, physical examination, US, Cystouretyhrography, and flowmetry. Thereafter 201 patients with the following conditions were selected for urodynamic assessment: vesicoureteral reflux, changes in bladder morphology (diverticulum, trabeculation), inadequate infundibulization of the bladder neck, diabetes, neurogenic bladder, obstructive pathology and bladders with a small volume capacity. The urodynamic study consisted of instantaneous voiding, cystomanometry and urethral pressure profile. RESULTS: Of these 201 patients, 45 had vesical hyperreflexia, 45 bladder disuse, 95 had a normal bladder, 12 had a hypoactive bladder and 2 had detrusor instability. CONCLUSION: The urodynamic study, apart from flowmetry, is basically indicated in patients awaiting renal transplantation who have a neurogenic bladder and/or suspected as having lower urinary tract obstruction. PMID- 9412363 TI - [Sphincter reeducation in non-coordinated urination syndrome]. AB - OBJECTIVE: The bladder re-education programs were initially associated with treatment of women with vesical instability. However, little has been published on the efficiency of the re-education techniques in the uncoordinated voiding syndrome of the Hinman syndrome. This study evaluated the urodynamic results of sphincter re-education in the treatment of the uncoordinated voiding syndrome. METHODS: Our series comprised 50 consecutive patients with uncoordinated voiding syndrome. The mean age was 11.9 years; 82% were females and 18% males. All patients underwent a complete urodynamic study with external perineal electromyography and videocystography. The sphincter re-education protocol was comprised of a manual pre-voiding training phase, an electromyographic pre voiding training phase and a voiding training phase. Upon completing the re education process, a new urodynamic study was performed and during the follow-up period, uroflowmetry studies were carried out with external perineal electromyography at 3, 6 and 12 months. Based on the response of the detrusor during voiding, we have classified the uncoordinated voiding syndrome into three types: type A, in which voiding is achieved through voluntary contraction of the detrusor; and type C, in which voiding is achieved through abdominal pressure. RESULTS: The clinical results obtained by sphincter re-education at one-year follow-up were: 42% cured, 22% improved, 2% relapsed and 14% showed no response. The remaining 20% were excluded from the treatment protocol. CONCLUSIONS: It is likely that the three types of the uncoordinated voiding syndrome are merely different physiopathological stages of the same vesico-urethral dysfunction. We consider sphincter re-education to be the treatment of choice for this type of patients. PMID- 9412364 TI - [Medical treatment of bladder instability. Our experience]. AB - OBJECTIVE: To identify the factors that influence response to treatment of vesical instability. METHODS: A retrospective study was conducted to assess the efficacy of drug therapy with oxybutinin and imipramine in 89 patients with urodynamically demonstrated detrusor hyperreactivity. Control evaluations were performed at 2, 5 and 8 months. Evaluation of the results took into account the etiology, pressure and volume at which the wave of instability appeared. RESULTS: The results were evaluated according to patient subjective criteria. We observed a positive response (cure and improvement) to treatment with oxybutinin alone or oxybutinin+imipramine in 66.25% of the cases; side effects were observed in 44%. There was a 20% improvement in the positive response rate when the wave intensity was greater than 55 cm H2O and the bladder volume at which this occurred was greater than 150 ml. No patient treated with second line drug therapy (flavoxate, nifedipine and trospium chloride) cured. CONCLUSIONS: The etiology of vesical instability did not influence response to therapy. Waves with a greater intensity and those that appeared at higher volumes responded better to treatment. Nearly half of the patients with side effects required a reduction of the dosage or withdrawal of the drug. Our results and those reported elsewhere indicate that non-responders to treatment with oxybutinin alone or in combination with imipramine are unlikely to improve with currently available drug therapy. PMID- 9412365 TI - [Changes in the urethral pressure profile after vaginal electrostimulation in the treatment of stress urinary incontinence]. AB - OBJECTIVE: To assess the changes in urethral pressure measurement after vaginal electrostimulation for the treatment of genuine urinary stress incontinence. METHODS: A clinical study, cystometry and urethral pressure profile were conducted on 15 female patients with urinary stress incontinence. RESULTS: The clinical results showed that electrostimulation was effective in 60% of the patients. The maximum urethral pressure and the functional urethral length increased after electrostimulation (from 47.72 +/- 16.47 cm H2O to 58.27 +/- 14.09 cm H2O, and from 3.9 +/- 1.56 cm to 5.25 +/- 1.69 cm, respectively). There was no relationship between the clinical improvement and the increment of both parameters. However, the increase of the maximum urethral pressure was higher in patients with improvement of incontinence (from 42.5 +/- 3.53 cm H2O to 49.65 +/- 11.26 cm H2O). CONCLUSIONS: Electrostimulation increases both the maximum urethral pressure and the functional urethral length. However, there is no relationship between these parameters and the effects of electrostimulation on urinary incontinence. PMID- 9412366 TI - [Simplified Ramirez urethropexy in the treatment of genuine stress urinary incontinence in women. Multicenter study of clinical and urodynamic results]. AB - OBJECTIVES: To evaluate the clinical and urodynamic data of a multicenter study on female urinary stress incontinence undergoing surgical repair with the Ramirez simplified urethropexy. METHODS: Clinical, urodynamic and videocystographic data were analyzed in a multicenter series of 340 female patients with urinary stress incontinence (mean age 51.7 +/- 9.7 years) before and after the Ramirez urethropexy technique (mean follow-up 21.7 months). RESULTS: Post surgical urinary continence was 78.4%. Cystocele repair was demonstrated in 57.7%. Urge incontinence decreased in 17.1%. Daytime frequency statistically significantly decreased in 19%. Urinary obstructive symptoms increased in 19.3%. Bladder instability significantly decreased posturethropexy. Peak urinary flow rate and mean urinary flow rate diminished in 65% and 59%, respectively. Postvoiding residual urine increased significantly. No statistical correlation between posturethropexy continence and videocystographic bladder neck morphology was observed. CONCLUSIONS: The clinical and urodynamic data obtained in our series indicate that the Ramirez urethropexy technique, a simple and fast procedure, may be considered an alternative treatment in female urinary stress incontinence. PMID- 9412368 TI - [Functional changes in the bladder of children with primary bladder diverticulum]. AB - OBJECTIVE: Primary vesical diverticulum is generally diagnosed during urological evaluation for recurrent urinary tract infection that are frequently associated with the symptoms of urinary dysfunction. A urodynamic study was performed in children with primary vesical diverticulum to confirm or discard the presence of vesicourethal dysfunction, its relationship with vesicoureteral reflux, which is associated with diverticulum, and the results achieved by surgical versus medical treatment. METHODS/RESULTS: A urodynamic study was carried out in 11 children (7 boys and 4 girls), aged 6 to 13 years, with primary vesical diverticulum. The diverticulum was parameatal in 7 cases (4 with reflux), posterolateral in 2 cases (1 with reflux), anterolateral in 1 patient and 1 patient had multiple bladder diverticula without neurological disorder. Surgery was indicated in 8 cases and 3 were managed conservatively. Vesicourethral function, at the time the diverticulum was diagnosed, was normal in 3 (27%) and pathological in 8 (73%) patients. The urodynamic study disclosed uncoordinated vesicosphincteric function (4 pts), vesical instability (3 pts) and vesical hypotony (1 pt). Of these 4 cases with urodynamic disorders, diverticulum was associated with vesicoureteral reflux. Of the 8 children submitted to surgery, 6 had a functional pathology preoperatively; following surgical treatment of the diverticulum, vesicourethral function returned to normal in all cases. Of the 3 cases managed conservatively, 2 had persistent urodynamic disorders despite drug therapy; one of them had clinical features of urinary incontinence. CONCLUSIONS: These fundings suggest that alterations of bladder function might arise from the anatomic changes in the detrusor muscle causing the diverticulum and contribute to the onset or persistence of reflux, and should be considered an additional criterion for indicating surgery. It is therefore suggested that a urodynamic study for its diagnosis be included in the protocol for evaluation of children with primary vesical diverticulum. PMID- 9412367 TI - [Female stress urinary incontinence. Treatment with collagen infiltration in the urethra]. AB - OBJECTIVES: To present the results obtained in patients with stress urinary incontinence treated with periurethral collagen injection. METHODS: 26 female patients with stress urinary incontinence were treated with bovine collagen injection; mean volume 10.8 cc. The results achieved by this therapeutic modality are described herein. RESULTS: Control evaluations performed during a period of one year showed highly satisfactory results had been achieved initially and the success rate gradually increased over the 12 months follow-up. Overall the final results showed a success rate of 34.6%, 38.4% showed frank improvement and 26.9% had a failed procedure. There were no significant differences in the results for both types of stress urinary incontinence. The results correlated with the severity of incontinence; the success rate was higher in the patients with low grade incontinence. CONCLUSIONS: Periurethral collagen injection is indicated in patients with type I and type III stress urinary incontinence who cannot benefit from surgery. Patients with type II stress urinary incontinence, however, do not benefit from this therapeutic modality. PMID- 9412369 TI - [Current overview of the treatment of postmyelomeningocele neurogenic bladder]. AB - OBJECTIVE: The present article reviews the current therapeutic modalities for neurogenic bladder secondary to myelomeningocele. METHODS: The procedures utilized in the treatment of neurogenic bladder are described. The results achieved by the artificial urinary sphincter in 43 patients are presented. RESULTS: Most of the patients were 8 to 20 years of age at the time the artificial urinary sphincter was implanted. A complementary surgical procedure was required in 11 patients and 10 have been included in the program of intermittent catheterization. Few complications have been observed. CONCLUSION: The implantation of an artificial sphincter can enhance the quality of life of incontinent patients that meet the appropriate clinical and urodynamic criteria. PMID- 9412370 TI - [Neurologic changes in patients with multiple sclerosis]. AB - OBJECTIVE: To evaluate the presence of urinary symptoms in 77 patients with multiple sclerosis. METHODS: The neurological compromise and the degree of functional disability were evaluated through the Minimum Dossier of Disability for Multiple Sclerosis. The pertinent data for the urinary symptoms were collected through a directed survey. A urodynamic study was performed to evaluate bladder function. RESULTS: 81.8% of the patients had urinary symptoms, the most prevalent being the mixed syndrome (52%). Neurological involvement and functional disability were greater in the male patients and there was a higher incidence of urinary symptoms (91%). Urgency (66.6%), frequency (60.3%) and dysuria (53.8%) were the most common urinary symptoms. The complication rate was low; infection was the most common complication (14.28%) and was more prevalent in the females. Detrusor hyperreflexia (60%) was the most frequent urodynamic finding. CONCLUSIONS: Urinary symptoms are frequent in multiple sclerosis (81.8%), the most prevalent being the mixed syndrome. Neurological involvement and functional disability are greater in the male patients and there is a higher incidence of urinary symptoms. There is a significant correlation between the severity of neurological compromise (pyramidal and cerebellar) and the degree of functional disability and the presence of urinary symptoms. PMID- 9412371 TI - [Current status of neurostimulation and neuromodulation for vesicourethral dysfunction]. AB - OBJECTIVE: To describe the current indications, techniques and results of sacral root stimulation in patients with spinal cord lesions as a treatment for patients with high pressure bladders and/or urinary incontinence despite conservative management, as well as sacral root neuromodulation with permanent stimulators for complex bladder dysfunction: vesical instability, sensory urgency, chronic pelvic pain and chronic voiding dysfunction. METHODS/RESULTS: The literature is reviewed, both techniques are described and the results of the most significant series are discussed, with special reference to the first groups that utilized these techniques. CONCLUSIONS: There is ample experience in the application of sacral root electrical stimulation. The reported results are comparable with those achieved by other treatments, such as augmentation cystoplasty. Neurostimulation and neuromodulation techniques are simple, the complications are minimal and they do not prelude the use of other therapies. PMID- 9412372 TI - [Neuroandrologic considerations in impotent patients with Peyronie's disease]. AB - OBJECTIVE: To asses the neuroandrologic profile of impotent patients with Peyronie's disease. METHODS: We conducted a pharmacological erection test and a study of the neuroandrologic profile of 13 impotent patients with Peyronie's disease. Eight patients hhad associated conditions and no associated disease was demonstrated in the remaining 5 patients. The neuroandrologic profile was based on bulbocavernous EMG, S2-S4 evoked potentials, somatosensorial potentials of pudendal nerve, cavernous smooth muscle electromyography (SPACE), sympathetic skin response, and cystometrogram. RESULTS: SPACE was altered in all the cases. In the patients with no associated disease, all other data of the neuroandrologic profile were normal. Patients with associated conditions demonstrated more alterations in all other data of the neuroandrologic profile and significant differences were observed in 7 cases. No differences in the type of alterations of SPACE were observed in 7 cases. No differences in the type of alterations of SPACE were observed between both groups. CONCLUSION: The impotence associated with Peyronie's disease could be due to an intrinsic lesion of the erectile smooth muscle. PMID- 9412374 TI - [Reflections of 2 patients with cancer of the prostate]. PMID- 9412373 TI - [IPSS in LUTS: a request for a correct terminology]. PMID- 9412375 TI - [Prostatic adenocarcinoma. A letter from a father to his unborn son]. PMID- 9412376 TI - [Distal detubularized sigmoidoplasty (Canarian neobladder). Midterm functional evaluation]. AB - OBJECTIVE: The aim of this report is to evaluate the medium term clinical and urodynamic results of the distal detubularized sigmoid neobladder, which we have called the "Canarian neobladder". METHODS: Since April, 1992, we have performed 26 orthotopic neobladder substitution using this particular technique. Ten patients were excluded from the study: two patients with early cancer progression; one female patient; one patient that had died a few days after the operation and 6 patients with less than 16 months follow-up. The remaining 16 patients were evaluated and had a mean follow-up of 22 months (range 6-54). We have performed clinical, urodynamic, radiographic and metabolic evaluation every six months during the first year and every 12 months thereafter. Clinical evaluation included daytime frequency, daytime and night-time urinary incontinence, urge and patient satisfaction with the clinical results. Flowmetry, cystometry and pressure-flow test were performed. The upper urinary tract was evaluated by renal ultrasound or pyelography. Entero-ureteral reflux was discarded by cystography. The International Continence Society terminology was utilized and clearly specified if otherwise. RESULTS: 87.5% of the patients were continent during the daytime and only two cases had diurnal urinary incontinence. Seventy-five percent of the patients had night-time incontinence and required external devices for its management. At two years follow-up, the mean diurnal frequency was 180 minutes and the mean bladder capacity was 338.2. First sensation was noted at 70.6% of bladder capacity. The neobladder pressure at maximum capacity during the filling phase was less than 30 cm H2O in all cases. Involuntary bladder contractions were demonstrated during cystomanometry in 93.7%. The pressure-flow test demonstrated neobladder contraction in 50% of cases and combined with abdominal straining in 50%. The foregoing allowed application of passive urethral resistance ratio (PURR) parameters, showing mean PURR initial values of 39.1 cm H2O and mean PURR curvature values of 0.17 cm H2O (mil/sec)2. The mean postvoiding urine was 37.5 ml. We diagnosed entero-ureteral reflux in 25% of patients and upper urinary tract dilatation due to a distal ureteral stricture in 2 patients. Mild outlet obstruction was diagnosed in 50% of the cases and were submitted to endoscopic section of the entero-urethral anastomosis. CONCLUSIONS: The distal sigmoid detubularized neobladder achieves a high rate of daytime continence with adequate frequency. However, the night-time incontinence rate was also high. The neobladder functional capacity remained stable throughout follow-up. Voiding was obtained as a result of neobladder contraction; thus postvoiding urine was minimal and patients did not require self catheterization. PMID- 9412377 TI - [Neurobiology of postoperative impotence after rectal excision]. AB - OBJECTIVE: The present study was conducted to determine the changes in the neuro andrologic profile of patients with impotence following rectal ablative surgery. METHODS/RESULTS: The study comprised 18 patients who had undergone rectal surgery: abdominoperineal resection of the rectum (AP) in 12 cases (67%), anterior resection of the rectum (AR) in 6 cases (33%). The pharmacologic erection test was negative in 60% of the patients (56% of the AP cases and 67% of the RA cases; differences not significant). Sympathetic lesion was demonstrated in 67% of the patients (50% of the AP cases and 100% of the AR cases; significant difference). Parasympathetic lesion was demonstrated in 38% of the patients (56% of the AP and in none of the RA cases; tendency towards statistical significance). Pudendal lesion was demonstrated in 83% of the patients, although no significant differences concerning pudendal involvement were observed between both types of surgery (92% of the AP group and 67% of the RA group). The frequency of the pudendal lesion was significantly greater than the parasympathetic lesion and the sympathetic lesion tended to be significantly greater than the parasympathetic lesion in patients undergoing ablative rectal surgery. No significant differences were observed between the pudendal and the sympathetic lesion in these patients. No relationship was observed between the type of neurologic lesion and the results of the pharmacologic erection test. CONCLUSIONS: The type of neurological lesion appears to be related with the level of the rectal surgery. The sympathetic innervation would be more frequently compromised in anterior resection of the rectum. The parasympathetic innervation would be more frequently compromised in abdominoperineal resection. The pudendal innervation would be affected by both types of surgical techniques. PMID- 9412378 TI - [Experience with radical prostatectomy as treatment of localized cancer of the prostate at the Fundacion Santa Fe de Bogota, Colombia]. AB - OBJECTIVE: To analyze the experience of the Fundacion Santa Fe de Bogota with radical prostatectomy in the treatment of localized prostatic cancer. METHODS: A retrospective study was conducted on 108 patients with localized carcinoma of the prostate stage T1-T2NxM0 submitted to radical prostatectomy from 1989 to 1994. RESULTS: Preoperatively, 50% of the patients had a PSA < 10 ng/ml and 14% had values that fell within the normal ranges of 0-4 mg/ml. A family history of prostate cancer was detected in 9.3% of the patients. The prostate cancer was clinically understaged in 75% of the patients, particularly those with stages T2a and T2b, and less significantly in those with stage T2c. Considering only those patients in whom the pathological staging had disclosed the cancer was not localized, this incidence accounted for 52% (n = 57). The presence of surgical margins was approximately 36%. The tumor recurrence rate was 26.9% and the complication rate was 6.8%. CONCLUSION: The relatively low complication rate in the present series shows that radical prostatectomy is a safe procedure that achieves good results if the cases are carefully selected and the diagnostic test are widely utilized. PMID- 9412379 TI - [Eosinophilic cystitis in children. Study of 4 cases]. AB - OBJECTIVE: To present our experience in the diagnosis and treatment of 4 children with eosinophilic cystitis. METHODS: 4 children (3 boys and 1 girl) with eosinophilic cystitis are described. The mean follow-up since diagnosis was 30 months, the mean age was 7 years and the main symptom was hematuria. The clinical features, etiology, course of the disease, pathological findings and diagnostic methods are reviewed. The definitive diagnosis was established by the pathological findings. None of the patients had a recurrence. CONCLUSIONS: The diagnosis of eosinophilic cystitis is exclusively based on the histological findings. There are no clinical or complementary pathognomonic signs. Like most authors, we believe that the immune system plays a decisive role in the appearance of this disease. PMID- 9412380 TI - [Wilms tumor treated with partial surgery. 31-year survival]. AB - OBJECTIVE: To report on a patient with Wilms' tumor treated by partial nephrectomy with 31 years survival. METHODS: Herein we describe a 33-year-old patient who had undergone surgery for a right renal tumor at age 2 years and 10 months. A partial nephrectomy was performed because the patient had left ureterohydronephrosis. Histological analysis of the surgical specimen disclosed a nephroblastoma or Wilms' tumor. RESULTS/CONCLUSIONS: The progressive deterioration of the left urinary tract, despite attempts to correct this condition, warranted its complete suppression. The patient has remained symptom free and leads an active social and working life 31 years after the first operation, which makes this an exceptional case in the world literature. PMID- 9412381 TI - [Use of Tc 99m DTPA in the follow-up of 2 pediatric patients diagnosed with megacalycosis or Puigvert's disease]. AB - OBJECTIVE: Megacalycosis or Puigvert's disease is a congenital anomaly of renal development characterized by caliceal dilatation, an increased number of calyces, associated with hypoplasia of the pyramids of Malpighi, and a normal renal pelvis. Renal function is always normal and there is no evidence of obstruction to urinary flow. The foregoing are important in distinguishing megacalycosis from congenital hydronephrosis. We have studied two pediatric patients with this renal anomaly by means of non invasive techniques, in order to demonstrate they had no urinary flow obstruction despite the caliceal dilatation. METHODS: Two cases of megacalycosis that had been diagnosed at our hospital from 1991 to 1995 are described and the literature is reviewed. Diagnosis was basically by intravenous urography. A diuretic renogram with technetium 99m diethylenetriaminepentaacetic acid (Tc 99m DTPA) showed no urinary flow obstruction in the anomalous kidney. This test was repeated several times, together with renal function studies like the technetium 99m dimercaptosuccinic acid scan (DMSA Scan). RESULTS: Both patients are asymptomatic and have a normally functioning kidney with no scars demonstrable on the DMSA scan. Renal urinary flow studies have remained within the normal ranges (elimination of more than 40% of the radionuclide 20 min after the administration of furosemide). US control evaluations have shown adequate renal growth and persistent caliceal dilatation in both cases. CONCLUSIONS: This congenital malformation must be considered when investigating renal dilatation, since megacalycosis does not require surgical treatment. Intravenous urography is useful in the diagnosis of this condition and Tc 99m DPTA is the best test for subsequent control evaluations. PMID- 9412382 TI - [Renal hematomas secondary to extracorporeal shockwave lithotripsy]. AB - OBJECTIVE: 0.6% of the patients submitted to extracorporeal shock wave lithotripsy (ESWL) present symptomatic renal hematoma that can permanently alter renal function and morphology. The present study analyzes the risk factors, types of renal hematoma secondary to shock waves and treatment. METHODS: The study comprised 9 patients with ESWL-induced renal hematoma that had been diagnosed by renal ultrasound and/or urography. Patient evaluation included CT and an isotopic renogram. Two patients had associated arterial hypertension and ischemic coronary heart disease treated with antiaggregants. In 4 patients the renal calculi had produced partial obstruction of the urinary tract. RESULTS: In 4 patients the ESWL-induced renal hematoma was less than 1/3 of the renal volume and did not alter renal function. These patients have been classified as having type I renal hematoma and have been successfully managed conservatively. In 5 patients the hematoma was more than 1/3 of the renal volume and renal function was impaired. These patients have been classified as having type II renal hematoma. Spontaneous reabsorption of the hematoma facilitated recovery of renal function in one patient, two had persistent hematoma and progressive renal function impairment, and the remaining two patients underwent delayed evacuation of the hematoma on days 30-45. Decompression renal surgery preserved renal function of the two kidneys. CONCLUSION: Advanced arteriosclerosis, arterial hypertension, primary coagulation disorders or secondary to antiaggregation therapy and obstruction of the urinary tract increase the risk of ESWL-induced renal hematoma. Renal hematoma following ESWL has been classified as type I or type II. In the latter, renal decompression surgery facilitates recovery of renal function. PMID- 9412383 TI - [Efficacy and complications of intraprostatic prosthesis in the treatment of benign hyperplasia of the prostate]. AB - OBJECTIVE: The UroLume stent has been utilized in the management of symptomatic benign prostatic hyperplasia (BPH), particularly in patients at high surgical risk. Our results are analyzed and the literature reviewed. METHODS: From June, 1994 to November, 1996, 19 patients at high surgical risk (ASA III/IV) with symptomatic BPH were treated at our center; 17 (89.4%) had an indwelling catheter. The UroLume stent was inserted under local anesthesia and intravenous sedation. RESULTS: Stent placement as monotherapy was found to be clinically effective in 15 patients (78.9%) with a mean follow-up of 8.2 +/- 5.7 months (range 1-18). The mean symptom score (Madsen-Iversen) was 6.1 +/- 1.2. Patients with no further manipulations had a mean maximum flow rate of 8.8 +/- 3.8 ml/s. Two patients (10.5%) required stent removal due to retrograde displacement in one case and perineal discomfort and irritative voiding symptoms in the other. In two other patients (10.5%), remnant tissue had to be resected in order to void spontaneously. Seventeen patients (89.4%) were satisfied with the results. No case of urosepsis or incrustations were documented. A microbiological analysis disclosed urinary tract infection in one patient. CONCLUSIONS: The UroLume stent is clinically effective in the treatment of symptomatic BPH in patients unfit for prostatic surgery. It carries a low incidence of major complications and procedure-related morbidity. PMID- 9412384 TI - [Echographic diagnosis of female urethral diverticulum]. AB - OBJECTIVE: To describe the utility of transrectal ultrasound as an alternative imaging technique in the diagnosis of diverticulum of the female urethra. METHODS/RESULTS: A 35-year-old female that had been initially diagnosed as having a benign tumor of the vagina is described. The voiding cystogram, positive pressure urethrography with a double balloon catheter and urethroscopy were falsely negative for urethral diverticulum of the female urethra. Subsequent evaluation by transrectal ultrasound disclosed on oval-shaped, anechoic lesion located posteriorly to base of the bladder. CONCLUSIONS: Transrectal ultrasound could be the diagnostic imaging technique of choice in patients suspected as having diverticulum of the female urethra. PMID- 9412385 TI - [Vesica ileale Padovana (VIP): surgical technique, long-term functional evaluation, complications and management]. AB - OBJECTIVE: An original technique for complete bladder replacement using an ileal segment--the VIP pouch--is described. The long-term functional results and early and late complications are presented. METHODS: The records of 209 VIPs performed following radical cystectomy for invasive bladder cancer from 1987 through 1995 were reviewed. Follow-up ranged from 6 to 87 months (mean: 24 mo.); the age of the patients ranged from 35 to 76 years (mean: 59.6 yrs.). RESULTS: 188 patients with a follow-up of at least 6 months have been evaluated. There was 1 postoperative death from massive pulmonary embolism, 23 pts. died from tumor progression and 4 are still alive with metastases. Early complications were observed in 10.5% of the pts., comprising 9 cases of prolonged ileus (5 functional and 4 obstructive), 3 pelvic hematomas, 2 deep venous thrombosis and 2 fistulas between the enteric anastomosis and the VIP pouch. Late complications were observed in 39.5%, in particular, 28 uretero-ileal stenosis (15%), 21 urethro-ileal stenosis (11%) and 15 laparoceles (8%). Clinically relevant metabolic disturbance has not been observed. Complete daytime continence was achieved in more than 90% of the cases and night-time continence was observed in 75% of the patients. The mean VIP manometric capacity was around 400 ml with low pressure during reservoir emptying; 39 patients (20%), showed voiding problems with a mean postmicturition residual of 150 ml (3 pts. require clean intermittent self-catheterization). CONCLUSIONS: VIP offers a simple and easy-to-perform surgical technique to provide a good capacity, low pressure, non refluxing reservoir employing only a 40 cm. ileal segment. The clinical and urodynamic results are good and offer high quality of life to patients undergoing cystectomy. The overall rate of late complications is fairly high, although conservative management is effective in most cases. PMID- 9412386 TI - [Giant hypertrophy of the prostate: 2,410 grams of weight and 24 cm in diameter]. AB - OBJECTIVE: To describe what we consider to be the largest prostatic hypertrophy reported in the literature. METHODS: A 57-year-old man consulted for prostatic symptoms during the last few months. He had a large palpable abdominal mass and DRE showed enlargement of the prostate. The intravenous urogram and CT scan were compatible with a retroperitoneal tumor and explorative laparotomy was performed. RESULTS: A large pelvic mass, dependent on the prostate, was found and resected. Pathological analysis of the surgical specimen disclosed a giant adenoma of the prostate, weighing 2410 gm and 24 cm in diameter. Immunohistochemical studies for PSA, vimentin, smooth actin and ki 67 were performed. The tumor showed glandular and stromal hyperplasia. CONCLUSION: Giant prostatic adenoma should be included in the differential diagnosis of retroperitoneal masses. To our knowledge, the case described herein is the largest prostatic hypertrophy reported in the literature. PMID- 9412387 TI - [Carcinoma of Bellini's ducts: apropos of a case]. AB - OBJECTIVE: To report an additional case of collecting duct carcinoma of the kidney, known as carcinoma of Bellini. METHODS/RESULTS: A 65-year-old male patient was admitted for left renal pain. An ultrasound scan showed a solid right renal mass. The patient underwent extended radical nephrectomy. Pathological analysis of the surgical specimen disclosed carcinoma of Bellini. The patient is asymptomatic 18 months postoperatively. CONCLUSIONS: Carcinoma of Bellini is an uncommon tumor type arising from the collecting duct cells and accounts for 0.5% 2% of all renal tumors. PMID- 9412388 TI - [Recurrent hydrocele in a patient with familial mediterranean fever]. AB - OBJECTIVE: To report an uncommon case of familial Mediterranean fever with urological manifestations. METHODS/RESULTS: A case of recurrent hydrocele in a patient with familial Mediterranean fever is described. CONCLUSIONS: Although familial Mediterranean fever is characterized by an acute febrile stage and involvement of the serosa, it may manifest as recurrent hydrocele due to involvement of the urological serosa, as in the case described herein. PMID- 9412389 TI - [Adrenal tuberculosis. Diagnosis using polymerase chain reaction]. AB - OBJECTIVE: To present a case of a patient suffering from adrenocortical insufficiency (Addison's disease) and to demonstrate the utility of the polymerase chain reaction (PCR) in the diagnosis of adrenal cortex tuberculosis. METHODS/RESULTS: Herein we describe a patient with Addison's disease who had developed pulmonary tuberculosis a few years earlier. CT and PCR test were utilized in making the diagnosis. CONCLUSION: Although adrenocortical tuberculosis is uncommon today, it must be considered when evaluating adrenocortical insufficiency, especially if the patient has or has had tuberculosis. Although Mycobacterium tuberculosis might not be demonstrated by bacteriologic techniques, PCR can be useful in making the etiological diagnosis. PMID- 9412390 TI - [Supraselective embolization of renal pseudoaneurysm caused by an abdominal stab wound]. AB - OBJECTIVE: To report the results of conservative management of post-traumatic (abdominal stab wound) pseudoaneurysm with arteriovenous fistula in segmental artery of left kidney and secondary persistent hematuria in an HIV positive patient. METHODS/RESULTS: A supraselective left renal arteriography revealed a pseudoaneurysm. We performed embolization of the aneurysmal artery with coils and fibrin particles. Resolution of the hematuria was immediately achieved with no side effects or significant complications. CONCLUSIONS: In our view, supraselective embolization is the first option in the treatment of post traumatic renal artery aneurysms due to its low morbidity and limited aggressiveness. PMID- 9412391 TI - [Bladder lithiasis formed from an IUD. A rare case]. AB - OBJECTIVE: To describe an uncommon case of bladder calculus arising from an intrauterine device (IUD). METHODS/RESULTS: A female patient with an IUD for the past 10 years consulted for frequent episodes of cystitis. Patient evaluation demonstrated a bladder calculus that warranted surgery. Treatment was by cystolithotomy, which disclosed a 6 cm calculus attached to the IUD. CONCLUSION: Bladder calculus arising from an IUD is rare. The case described herein was successfully managed by cystolithotomy. PMID- 9412392 TI - [Perineal tuberculous abscess. Presentation of a clinical case]. AB - OBJECTIVE: To describe a case of perineal abscess caused by Koch's bacillus in a patient with a history of bladder tumor. METHODS/RESULTS: The patient described herein had consulted at the emergency services for perineal abscess. He had previously undergone transurethral resection of a bladder tumor and had received prophylactic intravesical BCG instillation thereafter. A culture of the exudate led to the diagnosis of tuberculosis, with no microbiological relation to BCG therapy. During the course of the disease, the patient developed a urethrocutaneous fistula which resolved after specific medical treatment. CONCLUSIONS: In a patient with clinical features of tuberculosis and a recent history of exposure to BCG, it does not necessarily follow that this bacillus is the cause of the tuberculosis. Furthermore, not all tuberculous fistulas progress to chronicity or require surgical treatment. PMID- 9412393 TI - Databases and accountability. PMID- 9412394 TI - Severe deformity of a Palmaz-Schatz stent after normal surgical manipulation. PMID- 9412395 TI - Cardiovascular surgery, then and now. A festschrift in honor of Dr William H. Muller, Jr. PMID- 9412396 TI - [Pharmacoepidemiology of antibiotics in the treatment of respiratory tract infections in pediatric outpatients]. PMID- 9412398 TI - [Coamoxiclav in the empirical monotherapy in outpatients with community acquired pneumonia]. AB - Amoxyclav (amoxycillin/potassium clavulanate, A/PC) was used in a dose of 625 mg 3 times a day in the treatment of 68 outpatients at the age of 17 to 88 years (the average age of 49 years) with slight or moderate community-acquired pneumonia. In 49 per cent of the patients the pneumonia developed at the background of concomitant chronic diseases. The positive clinical effect was observed in 94 per cent of the patients. In 76 per cent of them a short-term treatment course of 5 days was sufficient. Before the treatment 91.8 per cent of the isolates proved to be susceptible to A/PC. The pathogen eradication after completion of the treatment was stated in 72 per cent of the cases. Moderate gastrointestinal adverse reactions developed in 4 patients (6 per cent). The results demonstrated high clinical and bacteriological efficacies of A/PC and made it possible to recommend the drug as the 1st class agent for the initial empirical therapy of community-acquired pneumonia in outpatients. PMID- 9412397 TI - [Amoxycillin/potassium clavulanate in inpatients with bronchopulmonary infections]. AB - Step-by-step therapy of patients with pneumonia and exacerbated chronic bronchitis with amoxyclav (amoxycillin/potassium clavulanate) in a dose of 1.2 g administered intravenously dropwise every 8 hours for the first 2 days of the treatment with subsequent oral use of the drug in a dose of 625 mg thrice a day for 5 days proved to be highly efficient. The recovery and improvement were stated in 19 (95 per cent) out of 20 patients. The adverse reaction (urticaria) was observed in 1 patient. Identical results were recorded in a comparative randomized trial with the use of cefotaxime in a dose of 1.0 g intramuscularly every 8 hours for 7 days. The pharmacoeconomic estimate showed the expediency of the step-by-step therapy with the use of amoxycillin/potassium clavulanate. PMID- 9412399 TI - [Empirical antibacterial therapy of community acquired pneumonia in outpatients]. PMID- 9412401 TI - [Isolation and biological characterization of Legionella pneumophila mutants resistant to aminoglycoside antibotics and their protective action]. AB - Preliminary estimation of virulence in some antibiotic resistant mutants of Legionella pneumophila, Philadelphia 1 in various models of infection revealed its decrease in the mutants resistant to azlocillin, cefotaxime, fluoroquinolone LIB-80, neamine and streptomycin. Detailed investigation of the neamine resistant mutants showed that in relation to streptomycin susceptibility such mutants could be divided into 3 classes: susceptible to streptomycin, resistant to high concentrations of streptomycin and resistant to moderate concentrations of streptomycin. Part of mutants Nea(r)Strr and Nea(r)Strr500 and all mutants Nea(r)Strr100 proved to be less virulent with respect to guinea pigs and chick embryos. The study of the spectinomycin resistant mutants of Legionella spp. did not reveal any changes in the virulence which made it possible to suggest that the influence of the mutations in the ribosomal protein genes determining resistance to streptomycin and neamine on virulence of L. pneumophila was based on the interdependence of the mutant effect on the suppression and the influence on the virulence detected by us in S. flexneri, Y. pseudotuberculosis, L. monocytogenes and F. tularensis. The Legionella mutants Nea(r)Strr100 were characterized by significant protective activity and protected immunized guinea pigs when tested in a model of their aerogenic infection. PMID- 9412400 TI - [Efficacy and tolerance of coamoxiclav in community-acquired lower respiratory tract infections in children]. AB - Amoxyclav (amoxycillin/potassium clavulanate, A/PC) was used in the treatment of 55 children with acute bronchitis and pneumonia. The drug was administered in a dose of 20-40 mg/kg body weight a day in 3 portions. The treatment course was 4 to 10 days. The treatment was performed under careful clinicoroent-genologic control. The clinical picture of the disease in the children was characterized by a moderate process which made it possible to treat the children as outpatients. The clinical efficacy amounted to 90.5 per cent. The isolates of Streptococcus pneumoniae, Streptococcus pyogenes, Staphylococcus aureus and Haemophilus influenzae proved to be susceptible to A/PC. It may be used as the 1st class agent in the treatment of children with lower respiratory tract infection. PMID- 9412402 TI - [Community-acquired pneumonia: Etiological diagnosis]. AB - Microbiological and immunoserological approaches were used in etiological diagnosis of community-acquired pneumonia. It was concluded that Streptococcus pneumoniae, Staphylococcus aureus, Pseudomonas aeruginosa, Legionella pneumophila and Klebsiella pneumoniae predominated in the etiological structure of present severe community-acquired pneumonia. The most actual causative agents of nonsevere community-acquired pneumonia in persons under 60 were S. pneumoniae, Hemophilus influenzae, Mycoplasma pneumoniae and Chlamydia pneumoniae. Nonsevere community-acquired pneumonia in persons over 60 and/ or at the background of chronic obstructive pulmonary diseases, diabetes mellitus or other affections was most frequently due to S. pneumoniae, H. influenzae and aerobic gramnegative microbes. PMID- 9412403 TI - [Drug effects informational services: databases for patenting and marketing]. AB - The review is concerned with databases (DB) for patent and commercial aspects of new drugs marketing. The DB characteristics are given and the search peculiarities in every DB and its possible access are discussed. BD accessible through the STN International are presented. PMID- 9412404 TI - [International collaborative surveillance study of lower respiratory tract pathogens (The Alexander Project)]. PMID- 9412405 TI - [Resistance to beta-lactams]. PMID- 9412406 TI - [Localization of rifampicin resistance determinants on genetic map of Saccharopolyspora erythraea]. AB - Genetic mapping of 2 mutations designated as rif5 and rif8 determining different levels of rifampicin resistance in Saccharopolyspora erythraea, an organism producing erythromycin, was performed. The mutations were inherited as chromosomal markers and localized in the same region of the chromosome restricted by ura1 and met1 loci. No close linking of rif5 and rif8 with any of the auxotrophic property markers in S. erythraea was detected. There were observed differences in the indices of correlation between the inheritance of rif8, rif5 and the chromosomal markers between which they were located. The data indicated that mutations rif5 and rif8 probably occurred in different genes. PMID- 9412407 TI - [Screening of drugs inhibiting cholesterol synthesis]. AB - The biosynthesis of fusidin, an antibiotic of steroid structure, was used as a model of steroid synthesis. Screening of compounds modifying the fusidin synthesis included 80 strains of mycelial fungi. A high frequency of such fungal cultures was observed. 9 cultures forming compounds which inhibited the synthesis of cholesterol in the cell culture of human hepatocytes were screened. A method for biological estimation of the activity of cholesterol synthesis inhibitors was developed on rabbits with high blood levels of cholesterol. It was shown that the cholesterol synthesis inhibitors, isolated as a result of the specific screening were not toxic and markedly lowered the cholesterol blood levels. PMID- 9412408 TI - [Antimicrobial effects of medicines which are not antibiotics]. AB - The data on antimicrobial activity of 69 different drugs not belonging to the class of typical antibiotics were examined. It was shown that many of them had antimicrobial activity against enterobacteria susceptible and resistant to antibiotics. Some of such drugs were able to eliminate the property of resistance to antibiotics and in particular that to chloramphenicol and tetracyclines. The data are indicative of the fact that many of the so called nonantibiotics have the capacity of active interference with the human microbial ecology. PMID- 9412409 TI - [Epidemiological situation in the Russian Federation in 1991-1996]. PMID- 9412410 TI - [Influence of difuracil (PAP-49) on normal intestinal microflora of rabbits. In vitro and in vivo experiments]. AB - The influence of difuracil (PAP-49), a novel nitrofuran, on saprotrophic and normal digestive tract microflora was studied in vitro and in vivo by comparison with that of furazolidone. It was shown that the minimum inhibitory concentration of both the drugs ranged from 0.01 to 1.56 micrograms/ml. The experiments on rabbits with the use of the drugs in doses of 5 and 10 mg/ kg body weight revealed that unlike furazolidone difuracil induced no noticable qualitative or quantitative changes in the intestinal microflora composition. PMID- 9412411 TI - [Development of resistance to difuracil (PAP-49) in bacteria and its activity in combination with other antibacterial drugs]. AB - Possible increasing of resistance to difuracil or N-(5-nitrofurfuryliden)-5 nitrofuran-2-(N'-acetyl) carboxamidohydrazon (PAP-49) in Staphylococcus sp., Streptococcus sp. and Escherichia coli was studied and the level of the resistance increasing was determined by the method of 30-fold passage of the cultures in subbacteriostatic concentrations of the drug. It was shown in vitro that the increase of the resistance to the new nitrofuran was insignificant. The most intensive adaptation was observed during the first 10 passages. During the following 10-15 passages the susceptibility level remained unchanged (E.coli) or was even somewhat higher (S. aureus and Streptococcus sp.). After that a new increase of the resistance was observed. The most marked changes in the susceptibility were detected in Streptococcus sp. and the least marked in Staphylococcus sp. and E.coli. Combinations of difuracil in subbacteriostatic concentrations with various antibacterial drugs provided both additive and synergistic effects with respect to S.epidermidis, S. pneumonase, E.coli and P.mirabilis. The most active combinations were those of difuracil with aminoglycoside antibiotics, penicillins or chloramphenicol. PMID- 9412412 TI - [Metronidazole effect on active oxygen production by human blood neutrophils]. AB - The in vitro effect of metronidazole on production of active oxygen by neutrophila and in the enzymatic system of glucose-glucose oxidase-peroxidase was studied by luminol-dependent chemiluminescence. An increase in the spontaneous and zymozan-stimulated chemiluminescence and a decrease in the phorbolmyristate acetate (PMA)-stimulated chemiluminescence after 2-hour preincubation of the neutrophils with 8.5 mM of metronidazole were observed. In concentrations of 0.9 to 8.7 mM metronidazole (without washing) dose-dependently lowered the neutrophil chemiluminescence in response to the effect of PMA and ionophore A23187 and to a lesser degree to that of zymozan. In doses of 20 to 100 mM the drug had an insignificant effect on production of active oxygen by the neutrophils in response to the cell stimulation by PMA, ionophore A23187 and zymozan. The data are in conformity with the scavenger effect of metronidazole on active oxygen radicals generating in the cell-free enzymatic system both in the presence and in the absence of superoxide dismutase. PMID- 9412413 TI - [A novel carbapenem active against imipenem resistant strains of Pseudomonas aeruginosa]. PMID- 9412414 TI - [Physiological role of extracellular proteases and calcium ions in the processes of differentiation and antibiotic production by Streptomyces albogriseolus 444]. AB - The extracellular proteolytic activity of Streptomyces albogriseolus 444 was investigated. S. albogriseolus 444 was shown to synthesize an extracellular proteolytic complex with metal-protease and trypsin-like activity defining differentiation of the substrate mycelium to the aerial one as well as the spore formation. The synthesis of the complex proceeded in the absence of the inductor in the medium i.e. constitutively. The complex heterogeneity was confirmed chromatographically. Low concentration of calcium ions (1.5 mM) stimulated the total caseinolytic activity and suppressed the activity of the trypsin-like proteases. PMID- 9412415 TI - [A system of step-by-step differentiation of methicillin-resistant Staphylococcus aureus]. AB - One hundred and eighty four strains of methicillin resistant Staphylococcus aureus (MRSA) isolated in pyosurgical and burn departments of Moscow, Minsk, Omsk, Tbilisi, Vologda, Smolensk and Dushanbe were differentiated with using phages of the International Set (BIS), two collections of experimental phages and two-probe fingerprinting. More than 50 per cent of the isolates could not be typed by the BIS phages. The Experimental Collection of the N.F. Gamaleya Research Institute of Epidemiology and Microbiology (Moscow) was more useful in the differentiation in comparison to the Experimental Collection of the London Health Centre. A new approach applied by us to the establishment of our collection i.e. screening of cultures with a definite specificity of the system of the restriction-modification (by a modified phage 85) followed by their differentiation with respect to the specificity of the prophages (by the induced phages with the respective modification specificity) provided reliable and reproducible results. Our collection made it possible to classify the phage type 85 strains which as was confirmed by the fingerprinting data belonged to 3 different genotype variants. The fingerprinting had no advantages over the typing by the phages of the more extended phage set and was recommended for differentiation of the strains (14 per cent) not sensitive to the phages used. A step-by-step scheme for typing MRSA easy for use in clinical and epidemiological laboratories is described. PMID- 9412416 TI - [Analysis of the etiologic structure of urinary tract infection and antibiotic resistance of its pathogens]. AB - The main pathogens of inflammatory diseases of the kidneys and upper urinary tracts in inpatients of an urological unit were gramnegative organisms of the family Enterobacteriaceae while the pathogens of the infection of the lower urinary tracts (nonspecific urethritis) and male genitalia were grampositive cocci. Pseudomonas aeruginosa, Enterobacter agglomerans and Proteus spp. (indole positive) were the chief causative agents of the hospital infections. The analysis of the materials revealed a tendency towards an increase in the microflora resistance to the most widely used antibiotics: aminoglycosides, cephalosporins and fluoroquinolones. This especially applied to the "problem" pathogen P.aeruginosa. Thus, in 1987 the portion of the P.aeruginosa gentamicin susceptible strains amounted to 52 per cent whereas in 1996 it was 13 per cent. The strains susceptible to ofloxacin equaled 79 per cent in 1988 and 44 per cent in 1995. At present the drugs of choice in the treatment of urinary tract infections due to P.aeruginosa are ceftazidime, cefpirome and amikacin (65, 64 and 62 per cent of the susceptible strains respectively). The importance of permanent microbiological monitoring and the respective correction of the therapy are indicated. PMID- 9412417 TI - [Determination of the nucleotide sequence of the silent aminoglycoside phosphotransferase gene in Streptomyces rimosus P3]. AB - The nucleotide sequence BgIII-Pstl of the DNA fragment containing the aph gene cloned from Streptomyces rimosus P3 not producing aminoglycoside antibiotics was determined. The aph gene was shown to encode neomycin phosphotransferase differing by the substrate specificity from the enzyme encoded by the aph genes from the organisms producing aminoglycoside antibiotics. Comparison of the cloned aph VIII gene and its product with the aph genes and their products from the antibiotic-producing actinomycetes and clinical microbial strains permitted to consider it as a representative of the third group genes which are likely widely distributed in soil microorganisms not producing aminoglycoside antibiotics. The gene length was 777 nucleotide pairs at the average GC content of 67 per cent. Comparison of the nucleotide and protein sequences of the S.rimosus gene with those of the genes cloned from the aminoglycoside-producing organisms (S.fradiae and Micromonospora chalcea) revealed high homology in the 3'-end region of the genes. However, the homology percentage by DNA for the S.rimosus gene was lower than that for the genes from the other organisms on their comparison with each other: 56-67 and 82-86 per cent respectively. Comparison of the profiles of the protein hydrophilic properties revealed differences in the region of the first 110 amino acids of the gene under the investigation. The amino acid sequence contained all the highly conservative regions characteristics of aminoglycoside phosphotransferases but had several amino acid substitutes detected for the first time including those in the region responsible for the antibiotic binding. PMID- 9412418 TI - [Etiological structure and antimicrobial susceptibility of Clostridium in war wound infection]. AB - The paper presents data on clinico-microbiological investigation of gunshot and firemine wounds in 27 injured during the Afghan war at the stage of their qualified surgical treatment 2-5 days after the injury. The quantitative and qualitative compositions of Clostridium that caused the infection and their susceptibility to 20 antimicrobials were determined. It was shown that the main pathogen of the anaerobic infection at the prehospital stage was Clostridium perfringens as a monoculture or association with aerobic and obligate anaerobic microbes. For rational antimicrobial therapy of gas gangrene it was advisable to preliminarily have antibioticograms of the isolates from the wounds. In the empirical therapy it was preferable to use penicillins (benzylpenicillin, ampicillin and others), rifampicin, metronidazole or ceftriaxone. The results are useful for substantiation of schemes for rational antimicrobial prophylaxis of wound infectious complications in injured in war time. PMID- 9412419 TI - [Amoxycillin/potassium clavulanate (augmentin)--current importance in the treatment of infection]. PMID- 9412420 TI - [Stents: experience at the Cardiology Hospital of the 21st century National Medical Center]. AB - From february 1995 to february 1997 we implanted 157 stents in 105 patients. Age ranged from 38 to 81 years (mean 58), there were 83 males and 22 females. In 62.8% cases we implanted one stent and in 39 (37.2%) cases 2 to 6. Eighty three were Palmaz/Schatz (P/S), 27 ACT-ONE, 18 Wiktor (W), 9 Gianturco Rubin (GR), 8 Wallstent, 6 XT-Bard and 6 microstent. Indication were de novo in 23.8%, 87.5% post failure PTCA and in 13.3% late PTCA restenosis. Implant was successful al 96.1% of the patients. The first 32 patients received oral anticoagulation, the last 72 received aspirin and ticlopidine only. COMPLICATIONS: 4.7% acute thrombosis, 0.9% sub-acute thrombosis, three of them (2.5%) developed myocardial infarction, 0.9% emergency surgical treatment, 2.8% vascular complications and death in 2.8%. During follow-up (1 to 18 months, mean 7.7) we repeated angiography and 35 patients two to 14 months (m = 5.6), 12 of them had restenosis, during dilation two cases had dissection of the main left coronary artery and were send to surgery, the others were dilated without complications. One case had restenosis of the stent with obstructive lesions in other vessels and was send to elective surgery. The reminded patients are symptom free and had negative stress test. We conclude that this technology is an excellent alternative to percutaneous myocardial revascularization. Larger trials with long term follow-up is necessary to determine the true incidence of restenosis with the different types of stents. PMID- 9412421 TI - [Simultaneous assessment of perfusion and myocardial viability with 2 isotope studies (thallium at rest and sestamibi in exercise). Initial experience in Mexico and Latin America]. AB - Rest-stress sestamibi single photon emission computed tomography (SPECT) has sensitivity and specificity similar to those of thallium 201 SPECT for detection of coronary artery disease. However, sestamibi is not ideal agent to study myocardial viability. There is not published experience in Latin American using dual isotope SPECT protocol to evaluate myocardial perfusion and viability. We studied 44 consecutive patients with coronary artery disease, 37 of them with previous myocardial infarction. Coronary angiography was performed in all patients. We used a 3 mCi rest T 201 SPECT followed by stress and 25 mCi sestamibi injection. Sestamibi SPECT was performed 30 minutes after exercise or 1 hour after pharmacologic stress with dipyridamole. To validate perfusion findings patients returning next day for rest sestamibi injection and SPECT. Scintigraphic data were read by two blinded expert using 20 SPECT segment analysis and each segment was scored using 5 points scoring system (0 = normal, 4 = absent uptake). The segmental score agreement between rest thallium 201 and rest sestamibi and the comparison of defect reversibility percentage and non reversibility between both protocols was 90.7%. CONCLUSION: Separate acquisition dual isotope myocardial perfusion SPECT is accurate for coronary artery disease evaluation. It showed a good agreement with rest-stress sestamibi SPECT for assessment of rest perfusion defects and reversibility and it was a better method to evaluate myocardial viability. PMID- 9412422 TI - [Echocardiographic findings in primary Sjogren syndrome]. AB - Primary Sjogren's syndrome is an autoimmune disorder characterized by an increased cellular and humoral activity, which determines immune-complex deposition at multisystemic level. The main morphologic and functional alterations associated with this syndrome at cardiovascular level have been described only in isolated cases. In this paper, 23 patients with primary Sjogren's syndrome were studied by transthoracic echocardiography. The aim of this study was to determine the relationship between the duration of the syndrome, sex and age with the type of cardiovascular abnormalities. All patients were women with mean age of 58 years (range from 39 to 76 years). The longest disease duration was 20 years and the shortest, 2 years. The main alterations were localized at valve level and are characterized by two patterns of thickening: 1) the first one involves the whole extension of one or more leaflets, 2) the other one is nodular and involves only the edge of one or more leaflets. The abnormal valves were mitral, aortic and tricuspid, but none of them showed a significant dysfunction. We did not find any association between the type of valve abnormalities and age or disease duration. It was concluded that the wide variety of morphologic abnormalities at valvular level were related with degenerative factors associated with the age in some cases, but in others its development probably depends on immunopathologic features of the primary Sjogren's syndrome. It must be proved in future studies whether the affected tissue can be assessed by immunohistochemical tests. PMID- 9412423 TI - [Treatment of acute myocardial infarction with rt-PA in 60 minutes. Cooperative study]. AB - Thrombolytic therapy (TT) modifies the natural history of acute myocardial infarction (AMI) diminishing morbi-mortality rate. In recent studies, modification of infusion velocity, decreased the mortality 10 percentage points. OBJECTIVE: Test if rt PA administration over an hour is safe and practical. MATERIAL AND METHODS: A prospective, cooperative trial during 3 years, included patients with AMI with less than 6 hours of the onset of symptoms that received rt-PA therapy. Initially 10 mg bolus and then 90 mg over 60 minutes period. Together with the administration of rt-PA, 5000 units of heparin was given, followed by 1000 units per hour adjusted to keep PTT at 1.5 to 2 times normal. All patients received aspirin and according of the evolution adjuvant therapy. We defined bleeding complications and/or cerebrovascular accident related to thrombolytic therapy. RESULTS: We included 225 patients who received rt-PA. Average age was 57.1 +/- 22.2 years, 78.7% males and 21.3% females. Arrival time at hospital was 2.93 +/- 1.7 hours. 82.2% were in class I-II by NYHA. 59.2% had anterior wall location and 32.4% posterior-inferior wall 80% had reperfusion criteria. Only 7.1% required transfusion and 0.4% presented CNS bleeding. The survival rate was 95.2%. The mortality had no relation with bleeding. CONCLUSION: Fast infusion is an effective and safe method. Transfusion requirements are no greater, and CNS bleeding was noted in 0.4% of the cases. PMID- 9412424 TI - [Incomplete ventricular laceration post mitral valve change. Report of a case and review of the literature]. AB - There are numerous reports concerning left ventricular rupture after mitral valve replacement, and it has been classified according to location, treatment and prognosis. A somewhat less terrific complication has appeared as modifications have been introduced to the surgical technique, decreasing the frequency of rupture, but otherwise increasing the frequency of incomplete laceration, which has seldom appeared in literature. We review the case of a patient with incomplete left ventricular laceration after mitral valve replacement. PMID- 9412426 TI - [Changes in variability of heart rate and sudden death detected during ambulatory electrocardiography]. AB - Two patients with ischemic heart disease had sudden death detected by means of ambulatory electrocardiography; the first patient had ventricular tachycardia asystole and the second bradycardia-asystole without mediating a ventricular tachyarrhythmia. In both cases the autonomic function of the heart was determined, by means of the heart rate variability for spectral analysis and for time domain analysis. An important decrease in heart rate variability in the patient with tachyarrhythmia was appreciated, more pronounced in the hour previous to his death. On the other hand, in the patient with bradycardia asystole there was increased heart rate variability. This suggests different neural influences in both patients for the development of sudden death: an alteration in the autonomic function of the heart in the patient with ventricular tachyarrhythmias, related to a depressed parasympathetic tone. In the patient with bradycardia-asystole, the parasympathetic tone was accentuated. PMID- 9412425 TI - [Identification and treatment of coronary atherosclerosis, before resection of abdominal aortic aneurysm]. AB - Atherosclerotic aortic aneurysm, is frequently associated to coronary atherosclerosis. When myocardial ischemia is asymptomatic, aortic surgery commonly is deferred because unexpected ischemic cardiopathy. To diminish the risk of aortic surgery, aortocoronary bypass must be installed before the aortic graft. Percutaneous transluminal coronary angioplasty is an alternative treatment of coronary atherosclerosis, principally in elderly patients. We present the case of a male patient with an abdominal aortic aneurysm and myocardial silent ischemia secondary to right coronary artery stenosis treated by mean the percutaneous transluminal coronary angioplasty (PTCA) before aortic surgery, with the objective of decreasing surgical risk and its possible complications (myocardial infarction, cardiogenic shock, death, etc.). Nine months after the PTCA, the patient is asymptomatic and stress test on treadmill is negative. PMID- 9412428 TI - [Indication for surgery in valvular diseases]. PMID- 9412427 TI - [Contributions of the most ancient medical schools to cardiology]. PMID- 9412429 TI - [Experimental infarction of the right ventricle. Its natural history]. AB - To elucidate the pathophysiology of severe right ventricular infarction (RVI), isolated RVI was produced in 12 dogs in an open chest preparation with intact pericardium. After RVI; mean right auricle pressure and right ventricular end diastolic pressure (RVEDP) increased (p < 0.05). A statistical significant decline was observed in cardiac output (43%, p < 0.05). However, for left ventricular systolic and end diastolic pressures (LVEDP) a decline was noted only after 120, or 90 min of the production of the RVI. Hemodynamic findings suggested a decreased preload of left chambers, as consequence of ischemic injury of the RV. This feature contributes to low cardiac output. A depression in right and left ventricular function curves were noted after RVI and this condition was maintained after 180 min of experimental observation. Equalized end-ventricular diastolic pressures were noted soon after RVI, a finding that was sustained over the time course of the experiment. Equalization of ventricular filling pressures was noted with RVEDP of < than 10 mmHg and with normals LVEDP. This features do not support, as a main cause, a restrictive role of the pericardium in the genesis of equalized filling pressures in RVI. PMID- 9412430 TI - [Analysis of reperfusion by thrombolysis of the artery responsible for acute myocardial infarction]. AB - OBJECTIVE: To analyze the role of the culprit coronary artery in myocardial infarction, its evolution and mortality. And to correlate with clinical criteria of reperfussion. MATERIALS AND METHODS: We included patients with clinical diagnosis of acute myocardial infarction (MI) treated with thrombolytic therapy, and coronariography. We used the TIMI study angiographic scale to evaluate the level of permeability of the culprit artery. RESULTS: Of 473 patients with of acute MI; coronariography was made in 377. The most frequent culprit vessel was anterior descending artery in 168 patients (45%) and right coronary artery in 139 patients (36%). In 276 patients the culprit vessel was permeable (73%). Of them in 30 patients, had TIMI 1 alterations, TIMI 2 in 97 patients, had TIMI 3 in 148 patients, only 102 patients had TIMI 0. In anterior MI the most frequent reperfussion arrhythmia was ventricular ectopic beats followed by slow ventricular tachycardia and ventricular tachycardia in 54%, ventricular fibrillation was observed only in six patients, of whom TIMI scale was 2 and 3 in five patients. In inferior MI, ventricular ectopic beats and slow ventricular tachycardia was seen in 25% of patients. In patients with permeable culprit artery we observed significant depression of ST segment, (159 patients, 42%), and significant increase in CK-MB levels, seen in 191 patients (51%). In the group of patients with total occlusion of the culprit artery, twenty-one (30%) had left ventricular disfuntion, and only six of them were in cardiogenic shock. In the group of patients with permeable culprit artery only two percent had cardiogenic shock. Therefore the analysis of the clinical evolution is the maia marker to take into consideration to send patients to early coronary arteriography with the objective to look for other therapeutic alternatives. PMID- 9412431 TI - [Asymptomatic ischemia in coronary arterial ectasia. Effects of anticoagulation]. AB - We studied 23 patients (22 men and 1 woman), their ages ranged from 31 to 71 years (55.9 +/- 9.7 years), with isolated coronary arterial ectasia. Seventeen patients presented angina pectoris, 19 had myocardial infarction. An angiographic image of intracoronary thrombus was observed in 5 patients. Before the administration of anticoagulants (oral warfarin) 16 patients showed unstable angina, the exercise EKG was positive in 9 patients, and 16 patients presented silent ischemia (showed by EKG-Holter), whose duration was 35.21 +/- 29.27 min per day. After anticoagulants, only 5 patients showed unstable angina pectoris. Exercise EKG was positive in 7 patients and only 7 patients showed silent ischemia, whose duration decreased significantly (P < 0.001) to 12.47 +/- 22.5 min per day. PMID- 9412432 TI - [Exercise test in young steers at 2800 M above sea level]. AB - Among its different activities the Mexican Health Ministry (SSA) promotes: 1) the specification of biological standards for the Mexican population; 2) mechanisms for early detection of physiopathological changes and 3) development of the appropriate technology as a basis for primary health care. As part of this program we studied a random sample of students at the "Colegio de Bachilleres" of Cuajimalpa a suburban area of Mexico City, situated at 2800 m above sea level, part of the equipment used was produced at the Center for Development and Technological Applications (CE-DAT), an agency of SSA. Height, weight, heart rate (HR), systolic and diastolic blood pressures (SBP/DBP) were measured at rest in sitting position, standing up and walking on a treadmill. The results show a population with similar physical characteristics to those described for other populations in Mexico, with HR values within a homogeneous distribution, except for three subjects with rates larger than 2 standard deviations (SD). At standing up, women showed an orthostatic reflex of 27 beats/min and men of 22 beats/min. At the beginning of the exercise, the HR curve becomes more homogeneous, showing a discrete ascending slope with a low correlation coefficient, suggesting low physical capacity of the studied population. As exercise continued, some subjects showed a low increment of HR, giving values differing by 2 SD. Five women and three men had resting SBP values lying 2 SD out of the mean values of the sample. During exercise, SBP increases 28 mmHg as average. Notwithstanding the low cardiac response, the time spent on the treadmill lies within the levels considered as good and, even, excellent, on the table of the National Institute of Cardiology. We discuss new procedures to analyze instantaneously the HR and the pressoreceptor reflex as well as their functional significance. PMID- 9412433 TI - [Fontan surgery in tricuspid atresia. Experience at the National Institute of Cardiology "Ignacio Chavez"]. AB - A retrospective analysis is presented of all patients under 18 years of age with tricuspid atresia and concordant ventriculo-arterial connection who underwent a Fontan procedure in the National Cardiology Institute "Ignacio Chavez", Mexico City, between January 1989 and December 1995. In this period Fontan procedure was performed in 22 patients with a mean age of 6.52 +/- 1.4 years. 14 of them were females. Thirteen patients had previous palliative procedure. Age of palliation was 10 +/- 12 months with period of 5 +/- 2 years between palliative and Fontan procedure. Overall mortality was 31% (7 patients); with a mortality of 42% between 1989 and 1992, of 22% between 1992 and 1995 and of 17% since 1994 with all 5 patients operated in 1995 surviving. Mean pulmonary pressure was 13.9 +/- 4.15 mmHg, mean pulmonary vascular resistance 1.66 +/- 0.57 U.W/m2, left ventricular end diastolic pressure 7.5 +/- 2.46 mmHg, ejection fraction 58 +/- 11% and Nakata and McGoon indexes 219 +/- 56 mm2/m2 and 2 +/- 0.1 respectively. No significant differences were found survivors and non-survivors, with exception of the left ventricular end-diastolic pressure (p < 0.05). Causes of death were multiple organ failure in 2, ventricular fibrillation in 3 and septic shock in 2. PMID- 9412434 TI - [Effectiveness of anticoagulant oral treatment in patients with thrombus in left ventricle after acute myocardial infarction]. AB - Left ventricular mural thrombi (LVMT) is a complication of acute myocardial infarction (AMI), that may produce peripheral embolism which could be fatal. In order to establish an adequate time of oral anticoagulant (OA) therapy, we undertook a prospective study that included 45 patients with AMI and left ventricular thrombi detected by echocardiographic study, in the first 5 to 10 days postinfarction, the study was repeated, in 3 and 6 months. Treatment with oral anticoagulant was initiated at the point of the detection of thrombi maintaining an INR of 1.5 to 2. Thirty nine patients (79%) were males and 6 (11%) were females, with an age of 29 to 85 years and a range of 62 +/- 11 years. Forty four patients (98%) presented anterior wall infarction and 1 (2%) posteroinferior infarction. In patients with anterior infarction, in 38 (85%) the thrombi was located at the apical wall (p < 0.05), 5 (11%) in the septal wall and other (2%) in anterior and apical walls. The patient with the posteroinferior infarction presented extension to the right ventricle, where the thrombus was located (2%). The contractility alterations related with thrombi were diskinesia, followed by hipokinesia and finally akinesia. The ejection fraction had not relationship with thrombi formation. LVMT dissolved in 32 patients (71%) at 3 months (p < 0.05), in 8 (18%) in 6 months and in 5 (11%) it was maintained for more than 6 months. None of the patients presented complications of OA. We conclude that the LVMT are more frequent in anterior infarctions, essentially in those that present diskinesia. The majority of LVMT are resolved in 6 months with OA therapy. PMID- 9412436 TI - [Vasovagal syncope. Management with propanthelin bromide]. AB - OBJECTIVE: To study propanthelin bromide efficacy in preventing vasovagal syncope relapse. SETTING: HGZ No. 3 IMSS, Mazatlan, Sinaloa, Mexico, from 1992 to 1995. PATIENTS: 10 patients with vasovagal syncope were selected from 41 syncope patients. DESIGN: Prospective longitudinal. MEASURES: clinical charts, neurologic and cardiologic evaluation, electrocardiogram, electroencephalogram, C.A.T., Holter, stress test and chest X rays were made. In 10 patients, 15 to 30 mg of propanthelin bromide thrice a day were administered, 12 month survey on was made measured. RESULTS: Of the 10 patients, 7 were women mean age 18 years. In 9 of them no recurrence was evident abandoned 1 treatment, 4 had side effects. CONCLUSIONS: Propanthelin bromide decreases vasovagal syncope episodes with few side effects. PMID- 9412435 TI - [Electrocardiographic data suggesting anteroseptal myocardial infarction, in presence of infarction of the left ventricle]. AB - We present a case of right ventricular myocardial infarction, secondary to postangioplasty occlusion of ventricular ramus of the right coronary artery, that developed electrocardiographic changes suggestive of septal myocardial infarction, this diagnosis was eliminated through angiographic study. We conclude that the carefully analysis of the electrocardiographic changes in ST segment in V1 to V4 can guide to the diagnosis of right ventricular myocardial infarction. For that reason we recommend the routinary register of the right electrocardiographic derivations as V3R and V4R, and left derivations V7 and V8, that is, the thoracic circle, in all patients with acute myocardial infarction regardless its location. PMID- 9412437 TI - [Assessment of ischemic cardiopathy with dynamic echocardiography]. PMID- 9412438 TI - [Contributions of Italian medical schools to cardiology]. PMID- 9412439 TI - [Guidelines and recommendations of the Mexican Society of Cardiology for the training in pediatric echocardiography]. PMID- 9412440 TI - [Effect of pentobarbital on ADP ribosylation]. AB - Pentobarbital, an inhibitor of respiratory chain at site II stimulates mitochondrial ADP ribosylation of glia cells. Results show an in cause of approximatively 30% of the ADP ribosyl transferase activity under pentobarbital treatment. ADP ribosyl transferase modifies two major mitochondrial proteins at molecular weight 50 and 80 KDa. The pentobarbital has been reported to induce neobiogenesis of mitochondria with enhancement of mitochondrial DNA synthesis. The stimulation of ADP ribosyl transferase activity under respiratory chain inhibition suggest the implication of this effect on neobiogenesis of mitochondria. PMID- 9412441 TI - [Immunochemical analysis of human monoclonal IGM with antibody cold agglutinin activity during Mycoplasma pneumoniae infection]. AB - Mycoplasma pneumoniae (MP) infection is associated with the emergence of anti-I cold agglutinins of IgM isotype, rarely complicated by hemolytic anemia. The mechanism by which the autoimmune disease occurs is still considerably controversed. We describe here a case of MP infection complicated by severe autoimmune hemolytic anemia with monoclonal anti-I IgM. IgM anti-I antibodies were purified by adsorption-elution on OI+ red cells. The red blood cell antibodies were found to be tightly associated with IgG and anti-MP specificity probably as immune complexes. The significance of these results is discussed. PMID- 9412443 TI - Expanding consulting skills. PMID- 9412444 TI - Haemorrhage from aortic wall granuloma following laparoscopic Nissen fundoplication. PMID- 9412442 TI - [Effect of olive leaves (Olea europea) feeding on the in vitro JH III biosynthesis by the corpora allata in Schistocerca gregaria during vitellogenesis]. AB - Per os administration of olive leaves (Olea europea) to females of Schistocerca gregaria results in stopping vitellogenesis. These vitellogenins are not synthesised by the fat body in the heamolymph. The vitellogenin inhibition is induced by the stopping of juvenile hormone JH III by the corpora allata. These corpora allata (Medicago sp.) Synthesise 10 times less JH III than those of alfalfa fed females. PMID- 9412445 TI - Gastric squamous cell carcinoma in three horses. AB - Gastric squamous cell carcinoma was diagnosed in three horses. Clinical signs observed in all cases were weight loss, anorexia and lethargy. Respiratory signs were prominent in one case. All three horses had depressed albumin and elevated globulin and fibrinogen concentrations. Two horses were mildly anaemic. Inflammatory exudates were present in peritoneal cavities in all cases, and cytological evaluation provided a positive diagnosis of squamous cell carcinoma in two cases. Pleural fluid samples taken from two cases were also classified as inflammatory exudates, but no neoplastic cells were detected on initial examination. In all cases the neoplasms had arisen from the oesophageal region of the stomach, and had metastised throughout the abdomen. Two cases had metastatic lesions within the pleural cavity. PMID- 9412446 TI - [Maternal transmission or bovine spongiform encephalopathy in the case of "Cindy" disproved]. AB - On December 27th, 1996 a Galloway cattle named "Cindy" died of bovine spongiform encephalopathy (BSE) in Hoxter. So far all cases of BSE reported in Germany have been imported from the UK. However, the identity and origin of "Cindy" was not clear. DNA sequence analysis of the mitochondrial D-loop region and DNA typing of micro-satellite sites finally revealed that "Cindy" was imported from the UK as well. PMID- 9412447 TI - [Small animal contribution to pediatric oncology]. AB - Retrospective analysis of biopsy material submitted to an Institute of Veterinary Pathology by private veterinary practitioners from 1993-1995 revealed tumors in 157 dogs and 71 cats under one year of age. Both histiocytomas and papillomas were excluded from the study. In the dog 55% (n = 87) and in the cat 73% (n = 52) of all tumors were diagnosed as malignant. In the dog tumors in order of frequency were: mammary neoplasia (n = 24, of those n = 10 malignant), soft tissue sarcomas (n = 24), hematopoietic tumors (n = 23), benign mesenchymal tumors (n = 16), mast cell tumors (n = 15), odontogenic tumors (n = 9), melanomas (n = 9, of those n = 5 malignant), osteosarcoma (n = 6), others (n = 31). In the feline, sarcomas were most common and of those hematopoietic tumors (lymphoma n = 23; 32%; mast cell tumors n = 12; 17%) were the two largest groups. The remaining sarcomas were 13 soft tissue sarcomas (18%) of which fibrosarcomas (n = 8) were most prevalent. Carcinomas where diagnosed in only 2 cases whereas among benign neoplasms (n = 19; 27%), epithelial tumors were the largest group (n = 14). In the order of frequency the following benign neoplasias were identified: fibroadenoma of mammary gland (n = 5), odontogenic tumors (n = 5), benign soft tissue tumors (n = 4), others (n = 6). PMID- 9412448 TI - [Air transportation of day-old chicks with regard to animal welfare]. AB - The transportation of chicks by aircraft increased in the last years greatly. High losses occurred frequently. A very important reason is water and food deprivation for too long a time between hatching and the arrival at the consignee. The process of transportation by aircraft, the development of the animal at hatching, and the clinical and chemical changes caused by water and food deprivation are described. In our opinion pathophysiological conditions are likely to appear 48 hours after hatching, when the animals are not fed and watered. It seems to be necessary to pay more attention to this problem. PMID- 9412449 TI - [Vulvar discharge before parturition and a risk factor for postpartum diseases of the sow and for early postnatal piglet losses]. AB - Periparturient biotechniques with prednisolon are widely used in Europe for the prevention of perinatal losses in intensive pig production. However, the routinely applied 100 mg prednisolon on the 113th day of pregnancy to the sow are not without controversy. In four intensive pig production units altogether 2143 sows treated thus were subjected to the evaluation of the following parameters: A: the presence or absence of vulvar discharge at the 110th day of pregnancy B: postparturient disease of the sow C: early postnatal piglet losses The results showed that the sows having prepartal vulvar discharge developed after prednisolon application significantly (p < 0.001) higher incidence of postparturient disease (20.5% versus 9.3%) when compared to the sows having no prepartal vulvar discharge. The early postnatal losses were significantly higher (p < 0.05) in sows having prepartal vulvar discharge and consecutive postparturient disease when compared to the healthy sows. It is the opinion of the authors that prepartal biotechnique with prednisolon (in order to reduce perinatal losses) in sows showing the signs of prepartal vulvar discharge is contraindicated. PMID- 9412450 TI - [Resistance to protein biosynthesis inhibitors in Staphylococci: resistance genes and their spread--a review]. AB - The rapid spread of antibiotic resistances in a wide variety of bacteria is mainly due to the location of antibiotic resistance genes on mobile genetic elements such as plasmids and transposons. Principal ways of transfer of plasmid- and transposon-encoded resistance genes are presented using examples of the predominant genes mediating resistances to protein biosynthesis inhibitors such as tetracyclines, aminoglycosides, macrolide-lincosamide-streptogramin B antibiotics, and chloramphenicol in staphylococci. Transfer between different staphylococcal cells is substantially based on transduction, transformation, conjugation and mobilization while transfer of resistance genes within the same bacterial cell often includes interplasmidic recombination events and chromosomal integration of resistance plasmids or transposons. The abilities of the transferred resistance plasmids or transposons to integrate or to be integrated into DNA molecules, plasmids or chromosomal DNA, of the new host cell are of major importance to circumvent strain-, species- or genusspecific barriers such as restriction/modification systems, plasmid incompatibilities or deficiencies of plasmid replication which may limit efficient resistance gene transfer. PMID- 9412451 TI - [Application of gonadotropin releasing hormone in dairy herds with fertility disturbances with regard to milk yield data and metabolic parameters]. AB - The presented study was made to determine the factors influencing the fertility supporting effect of GnRH application at the time of insemination in a dairy herd with fertility problems. The metabolic parameters considered influencing conception results were milk yield, milk protein and milk fat and for blood parameters Urea, GOT and Bilirubin. The total effect of GnRH consisted in an improved pregnancy rate by 18.8% compared to the non treated control group. The improvement of pregnancy rate was observed mainly in groups with an adequate supply of energy at the time of insemination. Therefore the use of GnRH at insemination time is more successful, when milking test data and blood parameters are taken into account and indicate an equilibrated supply in energy. To explain these effects endocrinological regulation models are discussed. PMID- 9412452 TI - [Detection and occurrence of verotoxin-forming and/or shigatoxin producing Escherichia coli (VTEC and/or STEC) in milk]. AB - Raw milk contaminated with VTEC was described as a source of human EHEC infection. Diagnosis of VTEC from milk is complicated by the low number of VT positive cells in the total bacterial count, the great variety of serovars with different combinations of virulence markers and the lack of characteristic biochemical properties for the cultural detection of all VTEC. The graduated procedure presented and used for the examination of milk samples is based on VT detection in suitable enrichment cultures and the selective isolation of VTEC by means of VT-specific monoclonal antibodies using the VT-colony immunoblot. This method was used to examine 127 samples of raw milk and 146 samples of certified raw milk (Vorzugsmilch) from 5 different regions in Germany. 3.9% of the raw milk samples and 2.1% of the certified raw milk samples were VTEC-positive. Except for one O157:H- isolate from a raw milk sample, the VTEC found belonged to the group of non-O157 VTEC. They were assigned to 5 different serovars with different combinations of virulence markers. Therefore, raw milk and certified raw milk will continue to present a potential source of EHEC infection. It is recommended to use the procedure presented for the elucidation of the route of infection and for the improvement of detection of VTEC and EHEC-strains in milk in order to obtain comparable data for diagnosis in the official food control laboratories of the federal lands. PMID- 9412453 TI - Generalization of the double-modulation method for in situ determination of elasticities. AB - The double-modulation method [Kacser and Burns (1979) Biochem. Soc. Trans. 7, 1149-1160] was the first method proposed for determining elasticities in situ. It is based on measuring changes in steady-state metabolite concentrations and fluxes induced by parameter modulations. It has the important advantage that it is not necessary to know the values of the changes in the parameters. Here we develop a matrix formulation of the double-modulation method that allows it to be applied to metabolic systems of any structure and size. It also shows which parameters need to be modulated and which variables need to be measured in order to calculate the elasticities that correspond to particular rates. Some suggestions for the practical implementation of the method are given, including various ways of testing the reliability of the results. PMID- 9412454 TI - Induction of subcutaneous tissue fibrosis in newborn mice by transforming growth factor beta--simultaneous application with basic fibroblast growth factor causes persistent fibrosis. AB - To establish an appropriate animal model of skin fibrosis by exogenous application of growth factors, we investigated the in vivo effects of transforming growth factor-beta by injection into subcutaneous tissue of newborn mice. Histological examination revealed that TGF-beta1, beta2, and beta3 induced granulation tissue formation after 3 days of injection, while these changes had disappeared after 7 days. The changes after 3 days of injection were more pronounced in the tissue injected with TGF-beta2 or beta3 than that with TGF beta1. In situ hybridization analysis indicated that connective tissue growth factor mRNA was strongly expressed in the fibroblasts at the site of TGF-beta injection, which suggested that fibroblasts were activated by TGF-beta. Next, we investigated the cooperative effects of TGF-beta and other growth factors including basic fibroblast growth factor (bFGF). The simultaneous application of TGF-beta and bFGF caused apparent tissue fibrosis which persisted for at least 2 weeks, while bFGF alone caused slight fibrotic changes after 7 days of injection. Thus, we succeeded in establishing an animal model of skin fibrotic disorders by the exogenous addition of growth factors, and this animal will be useful for future studies in this area. PMID- 9412455 TI - Cloning of P2Y6 cDNAs and identification of a pseudogene: comparison of P2Y receptor subtype expression in bone and brain tissues. AB - Cellular responses to ATP/UTP and analogs are mediated by G-protein coupled P2Y receptors and have been proposed to play a role in the regulation of bone metabolism. Using a degenerate PCR approach on MG-63 cell cDNA we found PCR fragments coding for human P2Y1 and a new receptor, P2Y6. cDNA cloning of the P2Y6 receptor identified three of cDNA isoforms. Two contained the same contiguous ORFs but differed in their 5 UTRs and may therefore originate by alternative splicing whereas the third represents a pseudogene. Analysis of P2Y receptor subtype expression in human bone and the osteoblastic cell lines OHS-4 and MG-63 by RT-PCR showed that all known human P2Y receptor subtypes (P2Y1, P2Y2, P2Y4, P2Y6, and P2Y7) were expressed. In contrast, analysis of brain derived cell lines suggests that a selective expression of P2Y receptor subtypes occurs in brain tissue. PMID- 9412456 TI - Spatial organization of large-scale chromatin domains in the nucleus: a magnified view of single chromosome territories. AB - We have analyzed the spatial organization of large scale chromatin domains in chinese hamster fibroblast, human lymphoid (IM-9), and marsupial kidney epithelial (PtK) cells by labeling DNA at defined stages of S phase via pulsed incorporation of halogenated deoxynucleosides. Most, if not all, chromosomes contribute multiple chromatin domains to both peripheral and internal nucleoplasmic compartments. The peripheral compartment contains predominantly late replicating G/Q bands, whereas early replicating R bands preferentially localize to the internal nucleoplasmic compartment. During mitosis, the labeled chromatin domains that were separated in interphase form a pattern of intercalated bands along the length of each metaphase chromosome. The transition from a banded (mitotic) to a compartmentalized (interphasic) organization of chromatin domains occurs during the late telophase/early G1 stage and is independent of transcriptional activation of the genome. Interestingly, generation of micronuclei with a few chromosomes showed that the spatial separation of early and late replicating chromatin compartments is recapitulated independently of chromosome number, even in micronuclei containing only a single chromosome. Our data strongly support the notion that the compartmentalization of large-scale (band size) chromatin domains seen in the intact nucleus is a magnified image of a similar compartmentalization occurring in individual chromosome territories. PMID- 9412457 TI - Lack of checkpoint control at the metaphase/anaphase transition: a mechanism of meiotic nondisjunction in mammalian females. AB - A checkpoint mechanism operates at the metaphase/anaphase transition to ensure that a bipolar spindle is formed and that all the chromosomes are aligned at the spindle equator before anaphase is initiated. Since mistakes in the segregation of chromosomes during meiosis have particularly disastrous consequences, it seems likely that the meiotic cell division would be characterized by a stringent metaphase/ anaphase checkpoint. To determine if the presence of an unaligned chromosome activates the checkpoint and delays anaphase onset during mammalian female meiosis, we investigated meiotic cell cycle progression in murine oocytes from XO females and control siblings. Despite the fact that the X chromosome failed to align at metaphase in a significant proportion of cells, we were unable to detect a delay in anaphase onset. Based on studies of cell cycle kinetics, the behavior and segregation of the X chromosome, and the aberrant behavior and segregation of autosomal chromosomes in oocytes from XO females, we conclude that mammalian female meiosis lacks chromosome-mediated checkpoint control. The lack of this control mechanism provides a biological explanation for the high incidence of meiotic nondisjunction in the human female. Furthermore, since available evidence suggests that a stringent checkpoint mechanism operates during male meiosis, the lack of a comparable checkpoint in females provides a reason for the difference in the error rate between oogenesis and spermatogenesis. PMID- 9412458 TI - Evidence for covalent modification of the nuclear dot-associated proteins PML and Sp100 by PIC1/SUMO-1. AB - PML and Sp100 proteins are associated with nuclear domains, known as nuclear dots (NDs). They were discovered in the context of leukemic transformation and as an autoantigen in primary biliary cirrhosis, respectively. Both proteins are expressed in the form of many COOH-terminally spliced variants, and their expression is enhanced by interferons (IFN). The recent finding that PIC1/SUMO-1, a small ubiquitin-like protein, is covalently linked to the RanGAP1 protein of the nuclear pore complex and also binds PML in yeast cells led us to determine whether PML is covalently modified by PIC1/SUMO-1 and whether the same is true for Sp100. We found an immune reaction of PML and Sp100 proteins with a PIC1/SUMO 1-specific monoclonal antibody by immunoblotting when using cell extracts prepared from stably transfected cells inducibly expressing one isoform of each protein as well as from nontransfected cells. In contrast, both proteins did not react when synthesized in vitro. Immunofluorescence staining showed that PIC1/SUMO-1 colocalized with Sp100 and PML in NDs except in mitotic cells, in which PML and Sp100 are dissociated. Cell fractionation and immunoblotting demonstrated that PIC1/SUMO-1 immunoreactive Sp100 in IFN-treated and untreated cells was exclusively nuclear, whereas nonmodified Sp100 was also found in the cytoplasm. Taken together, these data strongly suggest covalent modification of specific nuclear isoforms of Sp100 and PML by PIC1/SUMO-1. This modification may play a regulatory role in ND structure, composition, and function. PMID- 9412459 TI - Extracellular ATP activates transcription factor NF-kappaB through the P2Z purinoreceptor by selectively targeting NF-kappaB p65. AB - Cells of the macrophage lineage express a peculiar surface receptor for extracellular ATP, designated P2Z/P2X7 purinergic receptor, that induces pore formation and collapse of the plasma membrane potential. Although the function of the P2Z receptor is largely unknown, accumulating evidence implicates its role in cell signaling and immune reactions. Here, we investigated the effect of P2Z receptor ligation on the activation of NF-kappaB, a transcription factor controlling cytokine expression and apoptosis. Exposure of microglial cells to ATP but not other nucleotides resulted in potent NF-kappaB activation. This effect was specifically mediated by the P2Z receptor, because selective receptor antagonists prevented NF-kappaB activation. NF-kappaB activation required reactive oxygen intermediates and proteases of the caspase family, because it was abolished by antioxidants and specific protease inhibitors. The subunit composition of the ATP-induced NF- kappaB-DNA complex was rather unusual. Whereas exposure to LPS-induced prototypical NF-kappaB p50 homo- and p65 (RelA)/p50 heterodimers, ATP stimulation resulted in the sole appearance of a p65 homodimer. This is the first demonstration that a certain stimulus activates a particular NF kappaB subunit. Because different NF-kappaB complexes exhibit distinct transcriptional and DNA-binding activities, ATP may control the expression of a subset of NF-kappaB target genes distinct from those activated by classical proinflammatory mediators. PMID- 9412460 TI - A distinct and parallel pathway for the nuclear import of an mRNA-binding protein. AB - Three independent pathways of nuclear import have so far been identified in yeast, each mediated by cognate nuclear transport factors, or karyopherins. Here we have characterized a new pathway to the nucleus, mediated by Mtr10p, a protein first identified in a screen for strains defective in polyadenylated RNA export. Mtr10p is shown to be responsible for the nuclear import of the shuttling mRNA binding protein Npl3p. A complex of Mtr10p and Npl3p was detected in cytosol, and deletion of Mtr10p was shown to lead to the mislocalization of nuclear Npl3p to the cytoplasm, correlating with a block in import. Mtr10p bound peptide repeat containing nucleoporins and Ran, suggesting that this import pathway involves a docking step at the nuclear pore complex and is Ran dependent. This pathway of Npl3p import is distinct and does not appear to overlap with another known import pathway for an mRNA-binding protein. Thus, at least two parallel pathways function in the import of mRNA-binding proteins, suggesting the need for the coordination of these pathways. PMID- 9412461 TI - A nuclear import pathway for a protein involved in tRNA maturation. AB - A limited number of transport factors, or karyopherins, ferry particular substrates between the cytoplasm and nucleoplasm. We identified the Saccharomyces cerevisiae gene YDR395w/SXM1 as a potential karyopherin on the basis of limited sequence similarity to known karyopherins. From yeast cytosol, we isolated Sxm1p in complex with several potential import substrates. These substrates included Lhp1p, the yeast homologue of the human autoantigen La that has recently been shown to facilitate maturation of pre-tRNA, and three distinct ribosomal proteins, Rpl16p, Rpl25p, and Rpl34p. Further, we demonstrate that Lhp1p is specifically imported by Sxm1p. In the absence of Sxm1p, Lhp1p was mislocalized to the cytoplasm. Sxm1p and Lhp1p represent the karyopherin and a cognate substrate of a unique nuclear import pathway, one that operates upstream of a major pathway of pre-tRNA maturation, which itself is upstream of tRNA export in wild-type cells. In addition, through its association with ribosomal proteins, Sxm1p may have a role in coordinating ribosome biogenesis with tRNA processing. PMID- 9412463 TI - Analysis of the interactions of preproteins with the import machinery over the course of protein import into chloroplasts. AB - We have investigated the interactions of two nuclear-encoded preproteins with the chloroplast protein import machinery at three stages in import using a label transfer crosslinking approach. During energy-independent binding at the outer envelope membrane, preproteins interact with three known components of the outer membrane translocon complex, Toc34, Toc75, and Toc86. Although Toc75 and Toc86 are known to associate with preproteins during import, a role for Toc34 in preprotein binding previously had not been observed. The interaction of Toc34 with preproteins is regulated by the binding, but not hydrolysis of GTP. These data provide the first evidence for a direct role for Toc34 in import, and provide insights into the function of GTP as a regulator of preprotein recognition. Toc75 and Toc86 are the major targets of cross-linking upon insertion of preproteins across the outer envelope membrane, supporting the proposal that both proteins function in translocation at the outer membrane as well as preprotein recognition. The inner membrane proteins, Tic(21) and Tic22, and a previously unidentified protein of 14 kD are the major targets of crosslinking during the late stages in import. These data provide additional support for the roles of these components during protein translocation across the inner membrane. Our results suggest a defined sequence of molecular interactions that result in the transport of nuclear-encoded preproteins from the cytoplasm into the stroma of chloroplasts. PMID- 9412464 TI - Two distinct pathways for targeting proteins from the cytoplasm to the vacuole/lysosome. AB - Stress conditions lead to a variety of physiological responses at the cellular level. Autophagy is an essential process used by animal, plant, and fungal cells that allows for both recycling of macromolecular constituents under conditions of nutrient limitation and remodeling the intracellular structure for cell differentiation. To elucidate the molecular basis of autophagic protein transport to the vacuole/lysosome, we have undertaken a morphological and biochemical analysis of this pathway in yeast. Using the vacuolar hydrolase aminopeptidase I (API) as a marker, we provide evidence that the autophagic pathway overlaps with the biosynthetic pathway, cytoplasm to vacuole targeting (Cvt), used for API import. Before targeting, the precursor form of API is localized mostly in restricted regions of the cytosol as a complex with spherical particles (termed Cvt complex). During vegetative growth, the Cvt complex is selectively wrapped by a membrane sac forming a double membrane-bound structure of approximately 150 nm diam, which then fuses with the vacuolar membrane. This process is topologically the same as macroautophagy induced under starvation conditions in yeast (Baba, M., K. Takeshige, N. Baba, and Y. Ohsumi. 1994. J. Cell Biol. 124:903-913). However, in contrast with autophagy, API import proceeds constitutively in growing conditions. This is the first demonstration of the use of an autophagy like mechanism for biosynthetic delivery of a vacuolar hydrolase. Another important finding is that when cells are subjected to starvation conditions, the Cvt complex is now taken up by an autophagosome that is much larger and contains other cytosolic components; depending on environmental conditions, the cell uses an alternate pathway to sequester the Cvt complex and selectively deliver API to the vacuole. Together these results indicate that two related but distinct autophagy-like processes are involved in both biogenesis of vacuolar resident proteins and sequestration of substrates to be degraded. PMID- 9412462 TI - The Tim54p-Tim22p complex mediates insertion of proteins into the mitochondrial inner membrane. AB - We have identified a new protein, Tim54p, located in the yeast mitochondrial inner membrane. Tim54p is an essential import component, required for the insertion of at least two polytopic proteins into the inner membrane, but not for the translocation of precursors into the matrix. Several observations suggest that Tim54p and Tim22p are part of a protein complex in the inner membrane distinct from the previously characterized Tim23p-Tim17p complex. First, multiple copies of the TIM22 gene, but not TIM23 or TIM17, suppress the growth defect of a tim54-1 temperature-sensitive mutant. Second, Tim22p can be coprecipitated with Tim54p from detergent-solubilized mitochondria, but Tim54p and Tim22p do not interact with either Tim23p or Tim17p. Finally, the tim54-1 mutation destabilizes the Tim22 protein, but not Tim23p or Tim17p. Our results support the idea that the mitochondrial inner membrane carries two independent import complexes: one required for the translocation of proteins across the inner membrane (Tim23p Tim17p), and the other required for the insertion of proteins into the inner membrane (Tim54p-Tim22p). PMID- 9412466 TI - Ultrastructural organization of bovine chromaffin cell cortex-analysis by cryofixation and morphometry of aspects pertinent to exocytosis. AB - We have analyzed ultrathin sections from isolated bovine chromaffin cells grown on plastic support, after fast freezing, by quantitative electron microscopy. We determined the size and intracellular distribution of dense core vesicles (DVs or chromaffin granules) and of clear vesicles (CVs). The average diameter of DVs is 356 nm, and that of CVs varies between 35-195 nm (average 90 nm). DVs appear randomly packed inside cells. When the distance of the center of DVs to the cell membrane (CM) is analyzed, DV density is found to decrease as the CM is approached. According to Monte Carlo simulations performed on the basis of the measured size distribution of DVs, this decay can be assigned to a "wall effect." Any cortical barrier, regardless of its function, seems to not impose a restriction to a random cortical DV packing pattern. The number of DVs closely approaching the CM (docked DVs) is estimated to be between 364 and 629 (average 496), i.e., 0.45 to 0.78 DVs/micron2 CM. Deprivation of Ca2+, priming by increasing [Ca2+]i, or depolarization by high [K+]e for 10 s (the effect of which was controlled electrophysiologically and predicted to change the number of readily releasable granules [RRGs]) does not significantly change the number of peripheral DVs. The reason may be that (a) structural docking implies only in part functional docking (capability of immediate release), and (b) exocytosis is rapidly followed by endocytosis and replenishment of the pool of docked DVs. Whereas the potential contribution of DVs to CM area increase by immediate release can be estimated at 19-33%, that of CVs is expected to be in the range of 5.6-8.0%. PMID- 9412465 TI - Regulation of the ribosome-membrane junction at early stages of presecretory protein translocation in the mammalian endoplasmic reticulum. AB - A series of fusion protein constructs were designed to investigate the contribution of secretory nascent chains to regulation of the ribosome-membrane junction in the mammalian endoplasmic reticulum. As a component of these studies, the membrane topology of the signal sequence was determined at stages of protein translocation immediately after targeting and before signal sequence cleavage. Truncated translation products were used to delimit the analysis to defined stages of translocation. In a study of secretory protein precursors, formation of a protease-resistant ribosome-membrane junction, currently thought to define the pathway of the translocating nascent chain, was observed to be precursor- and stage-dependent. Analysis of the binding of early intermediates indicated that the nascent chain was bound to the membrane independent of the ribosome, and that the binding was predominately electrostatic. The membrane topology of the signal sequence was determined as a function of the stage of translocation, and was found to be identical for all assayed intermediates. Unexpectedly, the hydrophobic core of the signal sequence was observed to be accessible to the cytosolic face of the membrane at stages of translocation immediately after targeting as well as stages before signal sequence cleavage. Removal of the ribosome from bound intermediates did not disrupt subsequent translocation, suggesting that the active state of the protein-conducting channel is maintained in the absence of the bound ribosome. A model describing a potential mode of regulation of the ribosome-membrane junction by the nascent chain is presented. PMID- 9412467 TI - Cytoskeletal protein ABP-280 directs the intracellular trafficking of furin and modulates proprotein processing in the endocytic pathway. AB - Furin catalyzes the proteolytic maturation of many proproteins within the trans Golgi network (TGN)/endosomal system. Furin's cytosolic domain (cd) directs both the compartmentalization to and transit between its manifold processing compartments (i.e., TGN/biosynthetic pathway, cell surface, and endosomes). Here we report the identification of the first furin cd sorting protein, ABP-280 (nonmuscle filamin), an actin gelation protein. The furin cd was used as bait in a yeast two-hybrid screen to identify ABP-280 as a furin-binding protein. Binding analyses in vitro and coimmunoprecipitation studies in vivo showed that furin and ABP-280 interact directly and that ABP-280 tethers furin molecules to the cell surface. Quantitative analysis of both ABP-280-deficient and genetically replete cells showed that ABP-280 modulates the rate of internalization of furin but not of the transferrin receptor, a cycling receptor. However, although ABP-280 directs the rate of furin internalization, the efficiency of sorting of the endoprotease from the cell surface to early endosomes is independent of expression of ABP-280. By contrast, efficient sorting of furin from early endosomes to the TGN requires expression of ABP-280. In addition, ABP-280 is also required for the correct localization of late endosomes (dextran bead uptake) and lysosomes (LAMP-1 staining), demonstrating a pleiotropic role for this actin binding protein in the organization of cellular compartments and directing protein traffic. Finally, and consistent with the trafficking studies on furin, we showed that ABP-280 modulates the processing of furin substrates in the endocytic but not the biosynthetic pathways. The novel roles of ABP-280 and the cytoskeleton in the sorting of furin in the TGN/ endosomal system and the formation of proprotein processing compartments are discussed. PMID- 9412468 TI - Aggregation as a determinant of protein fate in post-Golgi compartments: role of the luminal domain of furin in lysosomal targeting. AB - The mammalian endopeptidase furin is a type 1 integral membrane protein that is predominantly localized to the TGN and is degraded in lysosomes with a t1/2 = 2-4 h. Whereas the localization of furin to the TGN is largely mediated by sorting signals in the cytosolic tail of the protein, we show here that targeting of furin to lysosomes is a function of the luminal domain of the protein. Inhibition of lysosomal degradation results in the accumulation of high molecular weight aggregates of furin; aggregation is also dependent on the luminal domain of furin. Temperature and pharmacologic manipulations suggest that furin aggregation occurs in the TGN and thus precedes delivery to lysosomes. These findings are consistent with a model in which furin becomes progressively aggregated in the TGN, an event that leads to its transport to lysosomes. Our observations indicate that changes in the aggregation state of luminal domains can be potent determinants of biosynthetic targeting to lysosomes and suggest the possible existence of quality control mechanisms for disposal of aggregated proteins in compartments of the secretory pathway other than the endoplasmic reticulum. PMID- 9412469 TI - Inhibition of endosome function in CHO cells bearing a temperature-sensitive defect in the coatomer (COPI) component epsilon-COP. AB - Recent evidence has suggested that subunits of the coatomer protein (COPI) complexes are functionally associated with endosomes in mammalian cells. We now provide genetic evidence that COPI plays a role in endocytosis in intact cells. The ldlF mutant CHO cell line bears a temperature-sensitive defect in the COPI subunit epsilon-COP. In addition to exhibiting conditional defects in the secretory pathway, we find that the cells are also defective at mediating endosome-associated functions. As found for cells microinjected with anti-COPI antibodies, ldlF cells at the restrictive temperature could not be infected by vesicular stomatitis (VSV) or Semliki Forest virus (SFV) that require delivery to acidic endosomes to penetrate into the cytosol. Although there was no temperature sensitive defect in the internalization of receptor-bound transferrin (Tfn), Tfn recycling and accumulation of HRP were markedly inhibited at the restrictive temperature. Sorting of receptor-bound markers such as EGF to lysosomes was also reduced, although delivery of fluid-phase markers was only partially inhibited. In addition, lysosomes redistributed from their typical perinuclear location to the tips of the ldlF cells. Mutant phenotypes began to emerge within 2 h of temperature shift, the time required for the loss of detectable epsilon-COP, suggesting that the endocytic defects were not secondary to a block in the secretory pathway. Importantly, the mutant phenotypes were also corrected by transfection of wild-type epsilon-COP cDNA demonstrating that they directly or indirectly reflected the epsilon-COP defect. Taken together, the results suggest that epsilon-COP acts early in the endocytic pathway, most likely inhibiting the normal sorting and recycling functions of early endosomes. PMID- 9412470 TI - The yeast adaptor protein complex, AP-3, is essential for the efficient delivery of alkaline phosphatase by the alternate pathway to the vacuole. AB - A novel clathrin adaptor-like complex, adaptor protein (AP)-3, has recently been described in yeast and in animals. To gain insight into the role of yeast AP-3, a genetic strategy was devised to isolate gene products that are required in the absence of the AP-3 mu chain encoded by APM3. One gene identified by this synthetic lethal screen was VPS45. The Vps pathway defines the route that several proteins, including carboxypeptidase Y, take from the late Golgi to the vacuole. However, vacuolar alkaline phosphatase (ALP) is transported via an alternate, intracellular route. This suggested that the apm3-Delta vps45 synthetic phenotype could be caused by a block in both the alternate and the Vps pathways. Here we demonstrate that loss of function of the AP-3 complex results in slowed processing and missorting of ALP. ALP is no longer localized to the vacuole membrane by immunofluorescence, but is found in small punctate structures throughout the cell. This pattern is distinct from the Golgi marker Kex2p, which is unaffected in AP-3 mutants. We also show that in the apm3-Delta mutant some ALP is delivered to the vacuole by diversion into the Vps pathway. Class E vps mutants accumulate an exaggerated prevacuolar compartment containing membrane proteins on their way to the vacuole or destined for recycling to the Golgi. Surprisingly, in AP-3 class E vps double mutants these proteins reappear on the vacuole. We suggest that some AP-3-dependent cargo proteins that regulate late steps in Golgi to vacuole transport are diverted into the Vps pathway allowing completion of transfer to the vacuole in the class E vps mutant. PMID- 9412471 TI - Cytoskeletal association is important for differential targeting of glucose transporter isoforms in Leishmania. AB - The major glucose transporter of the parasitic protozoan Leishmania enriettii exists in two isoforms, one of which (iso-1) localizes to the flagellar membrane, while the other (iso-2) localizes to the plasma membrane of the cell body, the pellicular membrane. These two isoforms differ only in their cytosolic NH2 terminal domains. Using immunoblots and immunofluorescence microscopy of detergent-extracted cytoskeletons, we have demonstrated that iso-2 associates with the microtubular cytoskeleton that underlies the cell body membrane, whereas the flagellar membrane isoform iso-1 does not associate with the cytoskeleton. Deletion mutants that remove the first 25 or more amino acids from iso-1 are retargeted from the flagellum to the pellicular membrane, suggesting that these deletions remove a signal required for flagellar targeting. Unlike the full length iso-1 protein, these deletion mutants associate with the cytoskeleton. Our results suggest that cytoskeletal binding serves as an anchor to localize the iso 2 transporter within the pellicular membrane, and that the flagellar targeting signal of iso-1 diverts this transporter into the flagellar membrane and away from the pellicular microtubules. PMID- 9412472 TI - The sleep-inducing lipid oleamide deconvolutes gap junction communication and calcium wave transmission in glial cells. AB - Oleamide is a sleep-inducing lipid originally isolated from the cerebrospinal fluid of sleep-deprived cats. Oleamide was found to potently and selectively inactivate gap junction-mediated communication between rat glial cells. In contrast, oleamide had no effect on mechanically stimulated calcium wave transmission in this same cell type. Other chemical compounds traditionally used as inhibitors of gap junctional communication, like heptanol and 18beta glycyrrhetinic acid, blocked not only gap junctional communication but also intercellular calcium signaling. Given the central role for intercellular small molecule and electrical signaling in central nervous system function, oleamide- induced inactivation of glial cell gap junction channels may serve to regulate communication between brain cells, and in doing so, may influence higher order neuronal events like sleep induction. PMID- 9412473 TI - Three-dimensional patterns and redistribution of myosin II and actin in mitotic Dictyostelium cells. AB - Myosin II is not essential for cytokinesis in cells of Dictyostelium discoideum that are anchored on a substrate (Neujahr, R., C. Heizer, and G. Gerisch. 1997. J. Cell Sci. 110:123-137), in contrast to its importance for cell division in suspension (DeLozanne, A., and J.A. Spudich. 1987. Science. 236:1086-1091; Knecht, D.A., and W.F. Loomis. 1987. Science. 236: 1081-1085.). These differences have prompted us to investigate the three-dimensional distribution of myosin II in cells dividing under one of three conditions: (a) in shaken suspension, (b) in a fluid layer on a solid substrate surface, and (c) under mechanical stress applied by compressing the cells. Under the first and second conditions outlined above, myosin II does not form patterns that suggest a contractile ring is established in the furrow. Most of the myosin II is concentrated in the regions that flank the furrow on both sides towards the poles of the dividing cell. It is only when cells are compressed that myosin II extensively accumulates in the cleavage furrow, as has been previously described (Fukui, Y., T.J. Lynch, H. Brzeska, and E.D. Korn. 1989. Nature. 341:328-331), i.e., this massive accumulation is a response to the mechanical stress. Evidence is provided that the stress-associated translocation of myosin II to the cell cortex is a result of the dephosphorylation of its heavy chains. F-actin is localized in the dividing cells in a distinctly different pattern from that of myosin II. The F actin is shown to accumulate primarily in protrusions at the two poles that ultimately form the leading edges of the daughter cells. This distribution changes dynamically as visualized in living cells with a green fluorescent protein-actin fusion. PMID- 9412474 TI - Myosin light chain-activating phosphorylation sites are required for oogenesis in Drosophila. AB - The Drosophila spaghetti squash (sqh) gene encodes the regulatory myosin light chain (RMLC) of nonmuscle myosin II. Biochemical analysis of vertebrate nonmuscle and smooth muscle myosin II has established that phosphorylation of certain amino acids of the RMLC greatly increases the actin-dependent myosin ATPase and motor activity of myosin in vitro. We have assessed the in vivo importance of these sites, which in Drosophila correspond to serine-21 and threonine-20, by creating a series of transgenes in which these specific amino acids were altered. The phenotypes of the transgenes were examined in an otherwise null mutant background during oocyte development in Drosophila females. Germ line cystoblasts entirely lacking a functional sqh gene show severe defects in proliferation and cytokinesis. The ring canals, cytoplasmic bridges linking the oocyte to the nurse cells in the egg chamber, are abnormal, suggesting a role of myosin II in their establishment or maintenance. In addition, numerous aggregates of myosin heavy chain accumulate in the sqh null cells. Mutant sqh transgene sqh-A20, A21 in which both serine-21 and threonine-20 have been replaced by alanines behaves in most respects identically to the null allele in this system, with the exception that no heavy chain aggregates are found. In contrast, expression of sqh-A21, in which only the primary phosphorylation target serine-21 site is altered, partially restores functionality to germ line myosin II, allowing cystoblast division and oocyte development, albeit with some cytokinesis failure, defects in the rapid cytoplasmic transport from nurse cells to cytoplasm characteristic of late stage oogenesis, and some damaged ring canals. Substituting a glutamate for the serine-21 (mutant sqh-E21) allows oogenesis to be completed with minimal defects, producing eggs that can develop normally to produce fertile adults. Flies expressing sqh-A20, in which only the secondary phosphorylation site is absent, appear to be entirely wild type. Taken together, this genetic evidence argues that phosphorylation at serine-21 is critical to RMLC function in activating myosin II in vivo, but that the function can be partially provided by phosphorylation at threonine-20. PMID- 9412475 TI - Actin-binding verprolin is a polarity development protein required for the morphogenesis and function of the yeast actin cytoskeleton. AB - Yeast verprolin, encoded by VRP1, is implicated in cell growth, cytoskeletal organization, endocytosis and mitochondrial protein distribution and function. We show that verprolin is also required for bipolar bud-site selection. Previously we reported that additional actin suppresses the temperature-dependent growth defect caused by a mutation in VRP1. Here we show that additional actin suppresses all known defects caused by vrp1-1 and conclude that the defects relate to an abnormal cytoskeleton. Using the two-hybrid system, we show that verprolin binds actin. An actin-binding domain maps to the LKKAET hexapeptide located in the first 70 amino acids. A similar hexapeptide in other acting binding proteins was previously shown to be necessary for actin-binding activity. The entire 70- amino acid motif is conserved in novel higher eukaryotic proteins that we predict to be actin-binding, and also in the actin-binding proteins, WASP and N-WASP. Verprolin-GFP in live cells has a cell cycle-dependent distribution similar to the actin cortical cytoskeleton. In fixed cells hemagglutinin-tagged Vrp1p often co-localizes with actin in cortical patches. However, disassembly of the actin cytoskeleton using Latrunculin-A does not alter verprolin's location, indicating that verprolin establishes and maintains its location independent of the actin cytoskeleton. Verprolin is a new member of the actin-binding protein family that serves as a polarity development protein, perhaps by anchoring actin. We speculate that the effects of verprolin upon the actin cytoskeleton might influence mitochondrial protein sorting/function via mRNA distribution. PMID- 9412477 TI - A novel family of serine/threonine kinases participating in spermiogenesis. AB - The molecular mechanisms regulating the spectacular cytodifferentiation observed during spermiogenesis are poorly understood. We have recently identified a murine testis-specific serine kinase (tssk) 1, constituting a novel subfamily of serine/threonine kinases. Using low stringency screening we have isolated and molecularly characterized a second closely related family member, tssk 2, which is probably the orthologue of the human DGS-G gene. Expression of tssk 1 and tssk 2 was limited to the testis of sexually mature males. Immunohistochemical staining localized both kinases to the cytoplasm of late spermatids and to structures resembling residual bodies. tssk 1 and tssk 2 were absent in released sperms in the lumen of the seminiferous tubules and the epididymis, demonstrating a tight window of expression restricted to the last stages of spermatid maturation. In vitro kinase assays of immunoprecipitates containing either tssk 1 or tssk 2 revealed no autophosphorylation of the kinases, however, they led to serine phosphorylation of a coprecipitating protein of approximately 65 kD. A search for interacting proteins using the yeast two-hybrid system with tssk 1 and tssk 2 cDNA as baits and a prey cDNA library from mouse testis, led to the isolation of a novel cDNA, interacting specifically with both tssk 1 and tssk 2, and encoding the coprecipitated 65-kD protein phosphorylated by both kinases. Interestingly, expression of the interacting clone was also testis specific and paralleled the developmental expression observed for the kinases themselves. These results represent the first demonstration of the involvement of a distinct kinase family, the tssk serine/threonine kinases, together with a substrate in the cytodifferentiation of late spermatids to sperms. PMID- 9412476 TI - Periplakin, a novel component of cornified envelopes and desmosomes that belongs to the plakin family and forms complexes with envoplakin. AB - The cornified envelope is a layer of transglutaminase cross-linked protein that is assembled under the plasma membrane of keratinocytes in the outermost layers of the epidermis. We have determined the cDNA sequence of one of the proteins that becomes incorporated into the cornified envelope of cultured epidermal keratinocytes, a protein with an apparent molecular mass of 195 kD that is encoded by a mRNA with an estimated size of 6.3 kb. The protein is expressed in keratinizing and nonkeratinizing stratified squamous epithelia and in a number of other epithelia. Expression of the protein is upregulated during the terminal differentiation of epidermal keratinocytes in vivo and in culture. Immunogold electron microscopy was used to demonstrate an association of the 195-kD protein with the desmosomal plaque and with keratin filaments in the differentiated layers of the epidermis. Sequence analysis showed that the 195-kD protein is a member of the plakin family of proteins, to which envoplakin, desmoplakin, bullous pemphigoid antigen 1, and plectin belong. Envoplakin and the 195-kD protein coimmunoprecipitate. Analysis of their rod domain sequences suggests that the formation of both homodimers and heterodimers would be energetically favorable. Confocal immunofluorescent microscopy of cultured epidermal keratinocytes revealed that envoplakin and the 195-kD protein form a network radiating from desmosomes, and we speculate that the two proteins may provide a scaffolding onto which the cornified envelope is assembled. We propose to name the 195-kD protein periplakin. PMID- 9412478 TI - Matrix metalloproteinase stromelysin-1 triggers a cascade of molecular alterations that leads to stable epithelial-to-mesenchymal conversion and a premalignant phenotype in mammary epithelial cells. AB - Matrix metalloproteinases (MMPs) regulate ductal morphogenesis, apoptosis, and neoplastic progression in mammary epithelial cells. To elucidate the direct effects of MMPs on mammary epithelium, we generated functionally normal cells expressing an inducible autoactivating stromelysin-1 (SL-1) transgene. Induction of SL-1 expression resulted in cleavage of E-cadherin, and triggered progressive phenotypic conversion characterized by disappearance of E-cadherin and catenins from cell-cell contacts, downregulation of cytokeratins, upregulation of vimentin, induction of keratinocyte growth factor expression and activation, and upregulation of endogenous MMPs. Cells expressing SL-1 were unable to undergo lactogenic differentiation and became invasive. Once initiated, this phenotypic conversion was essentially stable, and progressed even in the absence of continued SL-1 expression. These observations demonstrate that inappropriate expression of SL-1 initiates a cascade of events that may represent a coordinated program leading to loss of the differentiated epithelial phenotype and gain of some characteristics of tumor cells. Our data provide novel insights into how MMPs function in development and neoplastic conversion. PMID- 9412479 TI - The integrin alpha6beta4 functions in carcinoma cell migration on laminin-1 by mediating the formation and stabilization of actin-containing motility structures. AB - Functional studies on the alpha6beta4 integrin have focused primarily on its role in the organization of hemidesmosomes, stable adhesive structures that associate with the intermediate filament cytoskeleton. In this study, we examined the function of the alpha6beta4 integrin in clone A cells, a colon carcinoma cell line that expresses alpha6beta4 but no alpha6beta1 integrin and exhibits dynamic adhesion and motility on laminin-1. Time-lapse videomicroscopy of clone A cells on laminin-1 revealed that their migration is characterized by filopodial extension and stabilization followed by lamellae that extend in the direction of stabilized filopodia. A function-blocking mAb specific for the alpha6beta4 integrin inhibited clone A migration on laminin-1. This mAb also inhibited filopodial formation and stabilization and lamella formation. Indirect immunofluorescence microscopy revealed that the alpha6beta4 integrin is localized as discrete clusters in filopodia, lamellae, and retraction fibers. Although beta1 integrins were also localized in the same structures, a spatial separation of these two integrin populations was evident. In filopodia and lamellae, a striking colocalization of the alpha6beta4 integrin and F-actin was seen. An association between alpha6beta4 and F-actin is supported by the fact that alpha6beta4 integrin and actin were released from clone A cells by treatment with the F-actin- severing protein gelsolin and that alpha6beta4 immunostaining at the marginal edges of clone A cells on laminin-1 was resistant to solubilization with Triton X-100. Cytokeratins were not observed in filopodia and lamellipodia. Moreover, alpha6beta4 was extracted from these marginal edges with a Tween 40/deoxycholate buffer that solubilizes the actin cytoskeleton but not cytokeratins. Three other carcinoma cell lines (MIP-101, CCL-228, and MDA-MB-231) exhibited alpha6beta4 colocalized with actin in filopodia and lamellae. Formation of lamellae in these cells was inhibited with an alpha6-specific antibody. Together, these results indicate that the alpha6beta4 integrin functions in carcinoma migration on laminin-1 through its ability to promote the formation and stabilization of actin-containing motility structures. PMID- 9412480 TI - Effects of the metabotropic glutamate receptor antagonist MCPG on phosphoinositide turnover and synaptic plasticity in visual cortex. AB - The neurotransmitter glutamate, in addition to activating ligand-gated ion channels, also stimulates phosphoinositide (PI) hydrolysis in neurons by activating a group of G-protein-coupled metabotropic glutamate receptors (mGluRs). A role for mGluRs in synaptic plasticity originally was hypothesized based on the observation that the developmental decline in glutamate-stimulated PI turnover is well correlated with the decline in experience-dependent synaptic plasticity in visual cortex. Over the past few years, the compound alpha-methyl-4 carboxyphenylglycine (MCPG) has been widely used to test the role of PI-coupled mGluRs in a number of types of synaptic plasticity, including long-term potentiation (LTP), long-term depression (LTD), ocular dominance plasticity in visual cortex, and the neural plasticity underlying learning and memory. The conclusions of most of these studies were based on the assumption that MCPG blocks the actions of glutamate at PI-coupled mGluRs in the cerebral cortex. Here we show that this assumption is not valid in visual cortex. Although MCPG does antagonize the actions of the synthetic mGluR agonist 1S, 3R-aminocyclopentane 1,3-dicarboxylic acid, it fails to block PI turnover and changes in spike adaptation stimulated by glutamate, the endogenous mGluR ligand. In addition, we find that MCPG fails to block the NMDA receptor-dependent forms of LTP, LTD, and depotentiation in visual cortex. PMID- 9412481 TI - Input summation by cultured pyramidal neurons is linear and position-independent. AB - The role of dendritic morphology in integration and processing of neuronal inputs is still unknown. Models based on passive cable theory suggest that dendrites serve to isolate synapses from one another. Because of decreases in driving force or resistance, two inputs onto the same dendrite would diminish their joint effect, resulting in sublinear summation. When on different dendrites, however, inputs would not interact and therefore would sum linearly. These predictions have not been rigorously tested experimentally. In addition, recent results indicate that dendrites have voltage-sensitive conductances and are not passive cables. To investigate input integration, we characterized the effects of dendritic morphology on the summation of subthreshold excitatory inputs on cultured hippocampal neurons with pyramidal morphologies. We used microiontophoresis of glutamate to systematically position inputs throughout the dendritic tree and tested the summation of two inputs by measuring their individual and joint effects. We find that summation was surprisingly linear regardless of input position. For small inputs, this linearity arose because no significant shunts or changes in driving force occurred and no voltage-dependent channels were opened. Larger inputs also added linearly, but this linearity was caused by balanced action of NMDA and IA potassium conductances. Therefore, active conductances can maintain, paradoxically, a linear input arithmetic. Furthermore, dendritic morphology does not interfere with this linearity, which may be essential for particular neuronal computations. PMID- 9412482 TI - Regulation of Ca2+-dependent K+ channel expression in rat cerebellum during postnatal development. AB - Potassium channels govern duration and frequency of excitable membrane events and may regulate signals that are important in neuronal development. This study assesses the developmental expression of the large conductance Ca2+-dependent K+ channel in vivo and in vitro in rat cerebellum. In vivo, transcript levels for the Ca2+-dependent K+ channel (KCa) were shown by Northern analysis to increase during development, whereas transcript levels for the voltage-gated K+ channel Kv3.1, a delayed rectifier (KD), remained relatively constant. A comparable pattern was demonstrated by expression in Xenopus oocytes of poly(A)-enriched RNA isolated from postnatal rat cerebella. In cerebellar cultures, increased external K+ provided a simple manipulation of cell excitability that influenced KCa transcript levels during development. With low external K+ (5.3 mM), the levels of KCa channel transcript (assessed by semiquantitative PCR) remained constant throughout development. However, in culture medium that supported significant dendritic outgrowth (10 mM extracellular K+), an upregulation of KCa transcript level was observed similar to that seen in vivo. Tetraethylammonium (TEA; 1 mM) similarly enhanced KCa expression, suggesting that depolarizing stimuli increased KCa expression. The stimulatory effects of increased K+ or TEA on KCa expression required extracellular Ca2+ and were abolished in low external calcium (0.1 mM, buffered with EGTA), although morphological development and survival were not impaired. The regulation of KCa channel expression by depolarization and Ca2+ entry provides evidence of a logical feedback mechanism governing Ca2+ signals that may be significant in cerebellar development. PMID- 9412483 TI - Differential intracellular sorting of immediate early gene mRNAs depends on signals in the mRNA sequence. AB - This study characterizes the differential targeting of recently synthesized immediate early gene (IEG) mRNAs to neuronal cell bodies versus dendrites and tests the hypothesis that this targeting is based on signals in the encoded proteins. A single electroconvulsive seizure induces the expression of a number of IEG mRNAs in granule cells of the dentate gyrus. Most of these IEG mRNAs remain in the cell body, including two that are characterized in the present study (the mRNAs for NGFI-A and COX-2). In contrast, the mRNA for Arc moved rapidly into dendrites at an apparent rate of approximately 300 micron/hr. Inhibiting protein synthesis with cycloheximide did not disrupt the differential mRNA sorting, demonstrating that the differential targeting of mRNAs is not dependent on translation. PMID- 9412485 TI - Fluorescence-imaged microdeformation of the outer hair cell lateral wall. AB - Outer hair cell (OHC) electromotility appears to be central to mammalian hearing and originates within its lateral wall. The OHC lateral wall is a unique trilaminate structure consisting of the plasma membrane (PM), the cortical lattice (CL), and the subsurface cisternae (SSC). We selectively labeled and imaged the lateral wall components in the isolated guinea pig OHC under confocal microscopy. The PM was labeled with a voltage-sensitive dye, di-8-ANEPPS; the SSC was labeled with the sphingomyelin precursor, NBD-C6-ceramide; and F-actin in the CL was labeled with conjugates of phalloidin. Interactions among the three layers were evaluated with the micropipette aspiration technique. The PM was tethered to the CL and SSC until, at a critical deformation pressure, the PM separated, allowing visualization of the extracisternal space, and ultimately formed a vesicle. After detaching, the stiffness parameter of the PM was 22% of that of the intact lateral wall. We conclude that the lateral wall PM is more compliant than the CL/SSC complex. The data clarify the structural basis for electromotile force coupling in the OHC lateral wall. PMID- 9412484 TI - Potassium channel distribution, clustering, and function in remyelinating rat axons. AB - The K+ channel alpha-subunits Kv1.1 and Kv1.2 and the cytoplasmic beta-subunit Kvbeta2 were detected by immunofluorescence microscopy and found to be colocalized at juxtaparanodes in normal adult rat sciatic nerve. After demyelination by intraneural injection of lysolecithin, and during remyelination, the subcellular distributions of Kv1.1, Kv1.2, and Kvbeta2 were reorganized. At 6 d postinjection (dpi), axons were stripped of myelin, and K+ channels were found to be dispersed across zones that extended into both nodal and internodal regions; a few days later they were undetectable. By 10 dpi, remyelination was underway, but Kv1.1 immunoreactivity was absent at newly forming nodes of Ranvier. By 14 dpi, K+ channels were detected but were in the nodal gap between Schwann cells. By 19 dpi, most new nodes had Kv1.1, Kv1.2, and Kvbeta2, which precisely colocalized. However, this nodal distribution was transient. By 24 dpi, the majority of K+ channels was clustered within paranodal regions of remyelinated axons, leaving a gap that overlapped with Na+ channel immunoreactivity. Inhibition of Schwann cell proliferation delayed both remyelination and the development of the K+ channel distributions described. Conduction studies indicate that neither 4-aminopyridine (4-AP) nor tetraethylammonium alters normal nerve conduction. However, during remyelination, 4-AP profoundly increased both compound action potential amplitude and duration. The level of this effect matched closely the nodal presence of these voltage dependent K+ channels. Our results suggest that K+ channels may have a significant effect on conduction during remyelination and that Schwann cells are important in K+ channel redistribution and clustering. PMID- 9412487 TI - Synaptic transmission deficits in Caenorhabditis elegans synaptobrevin mutants. AB - Synaptobrevins are vesicle-associated proteins implicated in neurotransmitter release by both biochemical studies and perturbation experiments that use botulinum toxins. To test these models in vivo, we have isolated and characterized the first synaptobrevin mutants in metazoans and show that neurotransmission is severely disrupted in mutant animals. Mutants lacking snb-1 die just after completing embryogenesis. The dying animals retain some capability for movement, although they are extremely uncoordinated and incapable of feeding. We also have isolated and characterized several hypomorphic snb-1 mutants. Although fully viable, these mutants exhibit a variety of behavioral abnormalities that are consistent with a general defect in the efficacy of synaptic transmission. The viable mutants are resistant to the acetylcholinesterase inhibitor aldicarb, indicating that cholinergic transmission is impaired. Extracellular recordings from pharyngeal muscle also demonstrate severe defects in synaptic transmission in the mutants. The molecular lesions in the hypomorphic alleles reside on the hydrophobic face of a proposed amphipathic helical region implicated biochemically in interacting with the t-SNAREs syntaxin and SNAP-25. Finally, we demonstrate that double mutants lacking both the v SNAREs synaptotagmin and snb-1 are phenotypically similar to snb-1 mutants and less severe than syntaxin mutants. Our work demonstrates that synaptobrevin is essential for viability and is required for functional synaptic transmission. However, our analysis also suggests that transmitter release is not completely eliminated by removal of either one or both v-SNAREs. PMID- 9412486 TI - Developmental regulation of the cm2 muscarinic acetylcholine receptor gene: selective induction by a secreted factor produced by embryonic chick retinal cells. AB - The expression of the cm2 muscarinic acetylcholine receptor gene increases dramatically in chick retina during embryonic development in vivo. A similar developmental increase in cm2 expression occurs in embryonic chick retinal cells in culture. Conditioned medium from mature, but not young, retinal cultures contains a secreted factor that causes a selective increase in expression of cm2, but not cm3 or cm4, receptors. The secreted factor has been partially purified from serum-free medium, is protease-sensitive, and has a molecular weight >10 kDa. The cm2-inducing factor stimulates expression of a cm2 promoter/luciferase reporter gene, demonstrating that the increase in cm2 expression is attributable to increased gene transcription. Incubation of retinal cells with 14 identified neurotrophic and growth factors did not increase cm2 expression, suggesting that a novel developmentally regulated secreted factor mediates the subtype-specific induction of the cm2 receptor gene in retina. PMID- 9412488 TI - The relation of exocytosis and rapid endocytosis to calcium entry evoked by short repetitive depolarizing pulses in rat melanotropic cells. AB - Melanotropic cells release predocked, large, dense-cored vesicles containing alpha-melanocyte stimulating hormone in response to calcium entry through voltage gated calcium channels. Our first objective was to study the relationship between exocytosis, rapid endocytosis, and calcium entry evoked by short step depolarizations in the order of duration of single action potentials (APs). Exocytosis and rapid endocytosis were monitored by capacitance measurements. We show that short step depolarizations (40 msec) evoke the fast release of only approximately 3% of the predocked release-ready vesicle pool. Second, we asked what the distance is between voltage-gated calcium channels and predocked vesicles in these cells by modulating the intracellular buffer capacity. Exocytosis and rapid endocytosis were differentially affected by low concentrations of the calcium chelator EGTA. EGTA slightly attenuated exocytosis at 100 microM relative to 50 microM, but exocytosis was strongly depressed at 400 microM, showing that calcium ions have to travel a large distance to stimulate exocytosis. Nevertheless, the efficacy of calcium ions to stimulate exocytosis was constant for pulse durations between 2 and 40 msec, indicating that in melanotropes, exocytosis is related linearly to the amount and duration of calcium entry during a single AP. Rapid endocytosis was already strongly depressed at 100 microM EGTA, which shows that the process of endocytosis itself is calcium dependent in melanotropic cells. Furthermore, rapid endocytosis proceeded with a time constant of approximately 116 msec at 33 degrees C, which is three times faster than at room temperature. There was a strong correlation between the amplitude of endocytosis and the amplitude of exocytosis immediately preceding endocytosis. Both this correlation and the fast time constant of endocytosis suggest that the exocytotic vesicle is retrieved rapidly. PMID- 9412489 TI - Amphiphysin I antisense oligonucleotides inhibit neurite outgrowth in cultured hippocampal neurons. AB - Amphiphysin I is an SH3 domain-containing neuronal protein, enriched in axon terminals, which was reported to act as a physiological binding partner for dynamin I in synaptic vesicle endocytosis. Rvs167 and Rvs161, the yeast homologs of amphiphysin I, have been implicated in endocytosis, actin function, and cell polarity. Now we have explored the possibility that amphiphysin I also may have a role in actin dynamics and cell polarity by testing the effect of amphiphysin I suppression on neurite outgrowth. Freshly plated hippocampal neurons were exposed to antisense oligonucleotides via a new delivery system based on a polycationic amphipathic polymer, PS980. Western blot analysis revealed that amphiphysin I levels steadily increased with neuronal differentiation, whereas in antisense treated cultures amphiphysin I levels were reduced to approximately 10% of control levels at 48 hr. Concomitantly, a collapse of growth cones and a severe inhibition of neurite outgrowth and axon formation were observed. A similar effect was observed previously after dynamin I suppression in the same culture system (). We also have found that amphiphysin I and dynamin I colocalize in developing neurons at all developmental stages and that a pool of both proteins is colocalized with actin patches at the leading edge of growth cones. Our findings suggest a conserved role of the amphiphysin protein family in the dynamics of the cortical cell cytoskeleton and provide new evidence for a close functional link between amphiphysin I and dynamin I. PMID- 9412490 TI - Motoneuron apoptosis is blocked by CEP-1347 (KT 7515), a novel inhibitor of the JNK signaling pathway. AB - Neurons undergoing apoptosis can be rescued by trophic factors that simultaneously increase the activity of extracellular signal-regulated kinase (ERK) and decrease c-Jun N-terminal kinase (JNK) and p38. We identified a molecule, CEP-1347 (KT7515), that rescues motoneurons undergoing apoptosis and investigated its effect on ERK1 and JNK1 activity. Cultured rat embryonic motoneurons, in the absence of trophic factor, began to die 24-48 hr after plating. During the first 24 hr ERK1 activity was unchanged, whereas JNK1 activity increased fourfold. CEP-1347 completely rescued motoneurons for at least 72 hr with an EC50 of 20 +/- 2 nM. CEP-1347 did not alter ERK1 activity but rapidly inhibited JNK1 activation. The IC50 of CEP-1347 for JNK1 activation was the same as the EC50 for motoneuron survival. Inhibition of JNK1 activation by CEP-1347 was not selective to motoneurons. CEP-1347 also inhibited JNK1 activity in Cos7 cells under conditions of ultraviolet irradiation, osmotic shock, and inhibition of glycosylation. Inhibition by CEP-1347 of the JNK1 signaling pathway appeared to be selective, because CEP-1347 did not inhibit p38-regulated mitogen activated protein kinase-activated protein kinase-2 (MAPKAP2) activity in Cos7 cells subjected to osmotic shock. The direct molecular target of CEP-1347 was not JNK1, because CEP-1347 did not inhibit JNK1 activity in Cos7 cells cotransfected with MEKK1 and JNK1 cDNA constructs. This is the first demonstration of a small organic molecule that promotes motoneuron survival and that simultaneously inhibits the JNK1 signaling cascade. PMID- 9412491 TI - The mitogen-activated protein kinase p38-2 is necessary for the inhibition of N type calcium current by bradykinin. AB - Calcium influx via voltage-dependent calcium channels (ICa,V) links depolarization of excitable cells to critical cellular processes, such as secretion, contraction, and gene transcription. Fast regulation of ICa,V (<1 sec) by G-protein-coupled receptors is a relatively well-defined mechanism, whereas slow (30-60 sec) actions of transmitters and hormones on the same current remain poorly understood. In NG108-15 cells, the kinetically slow inhibition of N-type ICa,V by bradykinin (BK) requires the sequential activation of two G-proteins, heterotrimeric G13 and monomeric Rac1/Cdc42. We have now defined a role in this pathway for the relatively fast-acting p38 mitogen-activated protein kinase (MAPK). The slow inhibition of ICa,V by BK was suppressed specifically by SB203580, a compound that inhibits the p38 family of MAPKs. BK potently and selectively activated a newly discovered p38 family member, p38-2. These data provide the first evidence that a MAPK is involved in the regulation of ICa,V by a receptor-mediated process. PMID- 9412492 TI - Activation kinetics of AMPA receptor channels reveal the number of functional agonist binding sites. AB - AMPA and NMDA receptor channels are closely related molecules, yet they respond to glutamate with distinct kinetics, attributable to differences in ligand binding and channel gating steps (for review, see Edmonds et al., 1995). We used two complementary approaches to investigate the number of functional binding sites on AMPA channels on outside-out patches from cultured hippocampal neurons. The activation kinetics of agonist binding were measured during rapid steps into low concentrations of selective AMPA receptor agonists and during steps from a competitive AMPA receptor antagonist, 6-cyano-7-nitro-quinoxaline-2,3-dione, into a saturating concentration of agonist. Both approaches revealed sigmoidal kinetics, which suggests that multiple agonist binding steps or antagonist unbinding steps are needed for channel activation. A kinetic model with two independent binding sites gave a better fit to the activation phase than models with one or three independent sites. A more refined analysis incorporating cooperative interaction between the two binding sites significantly improved the fits to the responses. The affinity of the first binding step was two to three times higher than the second step. These results demonstrate that binding of two agonist molecules are needed to activate AMPA receptors, but the two binding sites are not identical and independent. Because NMDA receptors require four ligand molecules for activation (two glycine and two glutamate; Benveniste and Mayer, 1991; Clements and Westbrook, 1991), it may be that some binding sites on AMPA receptors are functionally silent. PMID- 9412493 TI - Interaction of muscle and brain sodium channels with multiple members of the syntrophin family of dystrophin-associated proteins. AB - Syntrophins are cytoplasmic peripheral membrane proteins of the dystrophin associated protein complex (DAPC). Three syntrophin isoforms, alpha1, beta1, and beta2, are encoded by distinct genes. Each contains two pleckstrin homology (PH) domains, a syntrophin-unique (SU) domain, and a PDZ domain. The name PDZ comes from the first three proteins found to contain repeats of this domain (PSD-95, Drosophila discs large protein, and the zona occludens protein 1). PDZ domains in other proteins bind to the C termini of ion channels and neurotransmitter receptors containing the consensus sequence (S/T)XV-COOH and mediate the clustering or synaptic localization of these proteins. Two voltage-gated sodium channels (NaChs), SkM1 and SkM2, of skeletal and cardiac muscle, respectively, have this consensus sequence. Because NaChs are sarcolemmal components like syntrophins, we have investigated possible interactions between these proteins. NaChs copurify with syntrophin and dystrophin from extracts of skeletal and cardiac muscle. Peptides corresponding to the C-terminal 10 amino acids of SkM1 and SkM2 are sufficient to bind detergent-solubilized muscle syntrophins, to inhibit the binding of native NaChs to syntrophin PDZ domain fusion proteins, and to bind specifically to PDZ domains from alpha1-, beta1-, and beta2-syntrophin. These peptides also inhibit binding of the syntrophin PDZ domain to the PDZ domain of neuronal nitric oxide synthase, an interaction that is not mediated by C-terminal sequences. Brain NaChs, which lack the (S/T)XV consensus sequence, also copurify with syntrophin and dystrophin, an interaction that does not appear to be mediated by the PDZ domain of syntrophin. Collectively, our data suggest that syntrophins link NaChs to the actin cytoskeleton and the extracellular matrix via dystrophin and the DAPC. PMID- 9412494 TI - Natural variation in neuron number in mice is linked to a major quantitative trait locus on Chr 11. AB - Common genetic polymorphisms-as opposed to rare mutations-generate almost all heritable differences in the size and structure of the CNS. Surprisingly, these normal variants have not previously been mapped or cloned in any vertebrate species. In a recent paper (), we suggested that much of the variation in retinal ganglion cell number in mice, and the striking bimodality of strain averages, are caused by one or two quantitative trait loci (QTLs). To test this idea, and to map genes linked to this variable and highly heritable quantitative trait, we have counted ganglion cells in 38 recombinant inbred strains (BXD and BXH) derived from parental strains that have high and low cell numbers. A genome-wide search using simple and composite interval-mapping techniques revealed a major QTL on chromosome (Chr) 11 in a 3 cM interval between Hoxb and Krt1 (LOD = 6.8; genome-wide p = 0.001) and possible subsidiary QTLs on Chr 2 and Chr 8. The Chr 11 locus, neuron number control 1 (Nnc1), accounts for one third of the genetic variance among BXH strains and more than half of that among BXD strains, but Nnc1 has no known effects on brain weight, eye weight, or total retinal cell number. Three strong candidate genes have been mapped previously to the same region as Nnc1. These genes-Rara, Thra, and Erbb2- encode receptors for retinoic acid, thyroxine, and neuregulin, respectively. Each receptor is expressed in the retina during development, and their ligands affect the proliferation or survival of retinal cells. PMID- 9412495 TI - Response of postmitotic neurons to X-irradiation: implications for the role of DNA damage in neuronal apoptosis. AB - The molecular changes responsible for inducing neuronal apoptosis are unknown. Rat cortical neurons were treated with x-irradiation 7 d after isolation to test for the role of DNA damage in neuronal death. The response of neurons to x irradiation was compared with that of astrocytes that had been isolated 3 weeks earlier from newborn rats. At the time of irradiation, the neurons appeared well differentiated morphologically and were predominantly (90-95%) noncycling, based on flow cytometric analysis. There was a similar, linear increase in DNA double strand breaks with increasing radiation dose in neurons and astrocytes. However, whereas doses as low as 2 Gy induced typical apoptotic changes in neurons, including nuclear fragmentation and/or internucleosomal DNA fragmentation, doses as high as 32 Gy caused little or no apoptosis in astrocytes. Radiation-induced apoptosis of neurons started 4-8 hr after irradiation, was maximal at 12 hr, and was dependent on dose up to 16 Gy. It was prevented when cycloheximide, a protein synthesis inhibitor, was added up to 6 hr after irradiation. In addition to their distinct apoptotic response, neurons rejoined radiation-induced DNA double-strand breaks more slowly than astrocytes. Treatment with benzamide to inhibit ADP ribosylation and strand break repair increased apoptosis; splitting the dose of radiation to allow increased time for DNA repair decreased apoptosis. These data suggest that DNA damage may induce neuronal apoptosis, that the extent of damage may determine the degree of apoptosis induced, and that slow repair of damage may play a role in the susceptibility of neurons to apoptosis. PMID- 9412496 TI - Neuroprotective effects of creatine and cyclocreatine in animal models of Huntington's disease. AB - The gene defect in Huntington's disease (HD) may result in an impairment of energy metabolism. Malonate and 3-nitropropionic acid (3-NP) are inhibitors of succinate dehydrogenase that produce energy depletion and lesions that closely resemble those of HD. Oral supplementation with creatine or cyclocreatine, which are substrates for the enzyme creatine kinase, may increase phosphocreatine (PCr) or phosphocyclocreatine (PCCr) levels and ATP generation and thereby may exert neuroprotective effects. We found that oral supplementation with either creatine or cyclocreatine produced significant protection against malonate lesions, and that creatine but not cyclocreatine supplementation significantly protected against 3-NP neurotoxicity. Creatine and cyclocreatine increased brain concentrations of PCr and PCCr, respectively, and creatine protected against depletions of PCr and ATP produced by 3-NP. Creatine supplementation protected against 3-NP induced increases in striatal lactate concentrations in vivo as assessed by 1H magnetic resonance spectroscopy. Creatine and cyclocreatine protected against malonate-induced increases in the conversion of salicylate to 2,3- and 2,5-dihydroxybenzoic acid, biochemical markers of hydroxyl radical generation. Creatine administration protected against 3-NP-induced increases in 3 nitrotyrosine concentrations, a marker of peroxynitrite-mediated oxidative injury. Oral supplementation with creatine or cyclocreatine results in neuroprotective effects in vivo, which may represent a novel therapeutic strategy for HD and other neurodegenerative diseases. PMID- 9412497 TI - Phosphorylation of mammalian olfactory cyclic nucleotide-gated channels increases ligand sensitivity. AB - In vertebrate olfactory sensory neurons, odorant receptors couple the sensory signal to the synthesis of the second messenger cAMP. Cyclic nucleotide-gated (CNG) channels are activated by binding of cAMP and conduct a depolarizing receptor current that leads to electrical excitation of the neuron. The sensitivity of olfactory CNG channels for cAMP can be significantly reduced by binding of calmodulin to a regulatory domain that resides within the N-terminus of the alpha-subunit of the channel. This regulatory domain also contains a consensus phosphorylation sequence for protein kinase C (PKC). We have investigated the effect of channel phosphorylation by PKC and found that phosphorylation increases ligand sensitivity without counteracting modulation of the channel by calmodulin. We have identified the amino acid residue that is phosphorylated by PKC and have localized three isoforms of PKC in olfactory sensory cilia. The results of this study provide information about the control of ligand sensitivity in olfactory CNG channels by an intrinsic regulatory domain, representing both a calmodulin-binding site and a substrate for PKC. PMID- 9412498 TI - Neurons produce a neuronal cell surface-associated chondroitin sulfate proteoglycan. AB - Monoclonal antibody Cat-315 recognizes a chondroitin sulfate proteoglycan (CSPG) expressed on the surface of subsets of neurons in many areas of the mammalian CNS (). The cell type-specific expression exhibited by the Cat-315 CSPG and other perineuronal net CSPGs imparts a distinct molecular surface identity to a neuron (Celio and Blumcke, 1994; Lander et al., 1997). The cell type(s) producing these surface-associated proteins and yielding this cellular diversity has remained in question. The expression of the Cat-315 CSPG in primary rat cortical cultures has permitted an examination of the cellular source of the Cat-315 antigen, as well as a determination of its spatial relationship to the neuronal surface. Live-cell labeling of primary neuronal cultures demonstrates that the Cat-315 CSPG is on the extracellular surface of neurons. Furthermore, extraction experiments demonstrate that the Cat-315 CSPG lacks a transmembrane domain and that the entire molecule is extracellular and, therefore, can be considered a constituent of brain extracellular matrix. Several lines of evidence indicate that neurons with cell surface staining produce the Cat-315 CSPG. First, neurons with cell surface staining also show intracellular Cat-315 immunoreactivity. Second, beta xyloside or monensin, reagents that inhibit the synthesis and transport of CSPGs, increase intracellular Cat-315 immunoreactivity within neurons that express cell surface Cat-315 immunoreactivity. Third, double labeling with Cat-315 and a polyclonal antibody for the Golgi complex demonstrates a precise colocalization of the intracellular Cat-315 immunoreactivity with the Golgi. Together, these observations demonstrate that neurons contribute to the extracellular matrix of brain and that the Cat-315 CSPG is produced by the neurons that carry Cat-315 cell surface immunoreactivity. PMID- 9412499 TI - Laminin directs growth cone navigation via two temporally and functionally distinct calcium signals. AB - During development, growth cones navigate to their targets via numerous interactions with molecular guidance cues, yet the mechanisms of how growth cones translate guidance information into navigational decisions are poorly understood. We have examined the role of intracellular Ca2+ in laminin (LN)-mediated growth cone navigation in vitro, using chick dorsal root ganglion neurons. Subsequent to contacting LN-coated beads with filopodia, growth cones displayed a series of stereotypic changes in behavior, including turning toward LN-coated beads and a phase of increased rates of outgrowth after a pause at LN-coated beads. A pharmacological approach indicated that LN-mediated growth cone turning required an influx of extracellular Ca2+, likely in filopodia with LN contact, and activation of calmodulin (CaM). Surprisingly, fluorescent Ca2+ imaging revealed no LN-induced rise in intracellular Ca2+ in filopodia attached to their parent growth cone. However, isolation of filopodia by laser-assisted transection unmasked a rapid, LN-specific rise in intracellular Ca2+ (+73 +/- 11 nM). Additionally, a second, sustained rise in intracellular Ca2+ (+62 +/- 8 nM) occurred in growth cones, with a distinct delay 28 +/- 3 min after growth cone filopodia contacted LN-coated beads. This delayed, sustained Ca2+ signal paralleled the phase of increased rates of outgrowth, and both events were sensitive to the inhibition of Ca2+/CaM-dependent protein kinase II (CaM-kinase II) with 2 microM KN-62. We propose that LN-mediated growth cone guidance can be attributed, in part, to two temporally and functionally distinct Ca2+ signals linked by a signaling cascade composed of CaM and CaM-kinase II. PMID- 9412500 TI - Isoform-specific effect of apolipoprotein E on cell survival and beta-amyloid induced toxicity in rat hippocampal pyramidal neuronal cultures. AB - Although the genetic link between the epsilon4 allele of apolipoprotein E (apoE) and Alzheimer's disease is well established, the isoform-specific activity of apoE underlying this correlation remains unclear. To determine whether apoE influences the neurotoxic actions of beta-amyloid (Abeta), we examined the effect of native preparations of apoE3 and E4 on Abeta-induced toxicity in primary cultures of rat hippocampal pyramidal neurons. The source of apoE was conditioned medium from HEK-293 cells stably transfected with human apoE3 or E4 cDNA. ApoE4 (10 microg/ml) alone was toxic to the cultures, whereas apoE3 had no effect. ApoE3 treatment prevented the toxicity induced by 10 microM Abeta(1-40) or Abeta(25-35). The apoE3 protective effect appears to be specific to Abeta-induced toxicity, because apoE3 did not protect against the cytotoxicity produced by NMDA or staurosporine, nor did apoE3 affect the increase in intracellular calcium induced by either NMDA or KCl. ApoE3 had no effect on the toxicity produced by Abeta in the presence of receptor-associated protein, an inhibitor of apoE receptors, particularly the LDL-receptor-related protein. Interaction with apoE receptors may not mediate the toxic actions of apoE4, because receptor-associated protein did not affect apoE4-induced neurotoxicity. Consistent with our previous biochemical experiments, analysis of the culture medium revealed that SDS-stable apoE3:Abeta complex is present in greater abundance than apoE4:Abeta complex. Thus, the protection from Abeta-induced neurotoxicity afforded by apoE3 treatment may result from clearance of the peptide by apoE3:Abeta complex formation and uptake by apoE receptors. PMID- 9412501 TI - Mitochondrial susceptibility to oxidative stress exacerbates cerebral infarction that follows permanent focal cerebral ischemia in mutant mice with manganese superoxide dismutase deficiency. AB - Mitochondrial injury has been implicated in ischemic neuronal injury. Mitochondria, producing adenosine triphosphate by virtue of electron flow, have been shown to be both the sites of superoxide anion (O2-) production and the target of free radical attacks. We evaluated these mechanisms in an in vivo cerebral ischemia model, using mutant mice with a heterozygous knock-out gene (Sod2 -/+) encoding mitochondrial manganese superoxide dismutase (Mn-SOD). Sod2 /+ mice demonstrated a prominent increase in O2- production under normal physiological conditions and in ischemia, as evidenced by specific oxidation of a fluorescent probe, hydroethidine, reflecting decreased activity of Mn-SOD. A mitochondrial viability assay that used rhodamine 123, which is accumulated by transmembrane potential of viable mitochondria, demonstrated accelerated development of mitochondrial injury. This rapid progress of ischemic injury resulted in exacerbation of infarct size and hemisphere enlargement, causing advanced neurological deficits but without altering DNA fragmentation induction. The present study suggests that O2- overproduced in a mitochondrial compartment, when uncoupled from antioxidant defenses, induces impairment of mitochondrial function and causes exacerbation of cerebral infarction after ischemia. PMID- 9412503 TI - Tissue and zonal-specific expression of an olfactory receptor transgene. AB - Discrimination of odorants is thought to arise from the selective expression of one of a small number of individual receptors in any single olfactory neuron. Receptor genes are expressed in a small subset of neurons throughout a zonally restricted region of the sensory epithelium. We demonstrate that a 6.7 kb region upstream of the M4 olfactory receptor coding region was sufficient to direct expression in olfactory epithelium. Moreover, reporter expression recapitulated the zonal restriction and distributed neuronal expression observed for endogenous olfactory receptors. Transgenic lines were obtained that directed expression in two different receptor zones, one of which was identical to the endogenous M4 receptor. When the reporter was expressed in the same zone as the endogenous M4 receptor, the two expression patterns were, in large part, nonoverlapping. These results suggest a model in which important regulatory elements are located in close proximity to transcription initiation sites of the olfactory receptor genes and receive information defining zonal patterning via long-range processes. PMID- 9412502 TI - In vivo expression and regulation of Elk-1, a target of the extracellular regulated kinase signaling pathway, in the adult rat brain. AB - The transcription factor Elk-1, a nuclear target of extracellular-regulated kinases (ERKs), plays a pivotal role in immediate early gene induction by external stimuli. Notably, the degree of phosphorylation of Elk-1 is tightly correlated with the level of activation of transcription of c-fos by proliferative signals. No data yet indicate the role of Elk-1 in the adult brain in vivo. To address this question, we have analyzed in the present work (1) Elk-1 mRNA and protein expression in the adult rat brain, and (2) the regulation of Elk 1 (i.e., its phosphorylation state) in an in vivo model of immediate early gene (IEG) induction: an electrical stimulation of the cerebral cortex leading to c fos and zif268 mRNA induction in the striatum. Using in situ hybridization, we show that Elk-1 mRNA is expressed in various brain structures of adult rat, and that this expression is exclusively neuronal. We demonstrate by immunocytochemistry using various specific Elk-1 antisera that the protein is not only nuclear (as shown previously in transiently transfected cell lines) but is also present in soma, dendrites, and axon terminals. On electrical stimulation of the glutamatergic corticostriatal pathway, we show a strict spatiotemporal correspondence among ERK activation, Elk-1 phosphorylation, and IEG mRNA induction. Furthermore, both activated proteins, analyzed by immunocytochemistry, are found in cytosolic and nuclear comparments of neuronal cells in the activated area. Our data suggest that the ERK signaling pathway plays an important role in regulating genes controlled by serum response element sites via phosphorylation of Elk-1 in vivo. PMID- 9412504 TI - Sox10, a novel transcriptional modulator in glial cells. AB - Sox proteins are characterized by possession of a DNA-binding domain with similarity to the high-mobility group domain of the sex determining factor SRY. Here, we report on Sox10, a novel protein with predominant expression in glial cells of the nervous system. During development Sox10 first appeared in the forming neural crest and continued to be expressed as these cells contributed to the forming PNS and finally differentiated into Schwann cells. In the CNS, Sox10 transcripts were originally confined to glial precursors and later detected in oligodendrocytes of the adult brain. Functional studies failed to reveal autonomous transcriptional activity for Sox10. Instead, Sox10 functioned synergistically with the POU domain protein Tst-1/Oct6/SCIP with which it is coexpressed during certain stages of Schwann cell development. Synergy depended on binding to adjacent sites in target promoters, was mediated by the N-terminal regions of both proteins, and could not be observed between Sox10 and several other POU domain proteins. Interestingly, Sox10 also modulated the function of Pax3 and Krox-20, two other transcription factors involved in Schwann cell development. We propose a role for Sox10 in conferring cell specificity to the function of other transcription factors in developing and mature glia. PMID- 9412505 TI - Sorting of beta-actin mRNA and protein to neurites and growth cones in culture. AB - The transport of mRNAs into developing dendrites and axons may be a basic mechanism to localize cytoskeletal proteins to growth cones and influence microfilament organization. Using isoform-specific antibodies and probes for in situ hybridization, we observed distinct localization patterns for beta- and gamma-actin within cultured cerebrocortical neurons. beta-Actin protein was highly enriched within growth cones and filopodia, in contrast to gamma-actin protein, which was distributed uniformly throughout the cell. beta-Actin protein also was shown to be peripherally localized after transfection of beta-actin cDNA bearing an epitope tag. beta-Actin mRNAs were localized more frequently to neuronal processes and growth cones, unlike gamma-actin mRNAs, which were restricted to the cell body. The rapid localization of beta-actin mRNA, but not gamma-actin mRNA, into processes and growth cones could be induced by dibutyryl cAMP treatment. Using high-resolution in situ hybridization and image-processing methods, we showed that the distribution of beta-actin mRNA within growth cones was statistically nonrandom and demonstrated an association with microtubules. beta-Actin mRNAs were detected within minor neurites, axonal processes, and growth cones in the form of spatially distinct granules that colocalized with translational components. Ultrastructural analysis revealed polyribosomes within growth cones that colocalized with cytoskeletal filaments. The transport of beta actin mRNA into developing neurites may be a sequence-specific mechanism to synthesize cytoskeletal proteins directly within processes and growth cones and would provide an additional means to deliver cytoskeletal proteins over long distances. PMID- 9412506 TI - Up and down states in striatal medium spiny neurons simultaneously recorded with spontaneous activity in fast-spiking interneurons studied in cortex-striatum substantia nigra organotypic cultures. AB - In vivo intracellular spontaneous activity in striatal medium spiny (MS) projection neurons is characterized by "up" and "down" states. How this type of activity relates to the neuronal activity of striatal fast-spiking (FS) interneurons was examined in the presence of nigral and cortical inputs using cortex-striatum-substantia nigra organotypic cultures grown for 45 +/- 4 d. The nigrostriatal projection was confirmed by tyrosine hydroxylase immunoreactivity. Corticostriatal (CS) projection neurons, striatal MS neurons, and FS neurons were intracellularly recorded and morphologically and electrophysiologically characterized. Intracellular spontaneous activity in the cultures consisted of intermittent depolarized periods of 0.5-1 sec duration. Spontaneous depolarizations in MS neurons were restricted to a narrow membrane potential range (up state) during which they occasionally fired single spikes. These up states were completely blocked by the glutamate antagonist CNQX. In FS interneurons, depolarized periods were characterized by large membrane potential fluctuations that occupied a wide range between rest and spike threshold. Also, FS interneurons spontaneously fired at much higher rates than did MS neurons. Simultaneous intracellular recordings established that during spontaneous depolarizations MS neurons and FS interneurons displayed correlated subthreshold neuronal activity in the low frequency range. These results indicate that (1) the CS projection neurons, striatal MS neurons, and FS interneurons grown in cortex striatum-substantia nigra organotypic cultures show morphological and electrophysiological characteristics similar to those seen in vivo; (2) striatal MS neurons but not FS interneurons show an up state; (3) striatal MS neurons and FS interneurons receive common, presumably cortical inputs in the low frequency range. Our results support the view that the cortex provides a feedforward inhibition of MS neuron activity during the up state via FS interneurons. PMID- 9412507 TI - Mouse Zic1 is involved in cerebellar development. AB - Zic genes encode zinc finger proteins, the expression of which is highly restricted to cerebellar granule cells and their precursors. These genes are homologs of the Drosophila pair-rule gene odd-paired. To clarify the role of the Zic1 gene, we have generated mice deficient in Zic1. Homozygous mice showed remarkable ataxia during postnatal development. Nearly all of the mice died within 1 month. Their cerebella were hypoplastic and missing a lobule in the anterior lobe. A bromodeoxyuridine labeling study indicated a reduction both in the proliferating cell fraction in the external germinal layer (EGL), from 14 d postcoitum, and in forward movement of the EGL. These findings suggest that Zic1 may determine the cerebellar folial pattern principally via regulation of cell proliferation in the EGL. PMID- 9412508 TI - Blockade and recovery of spontaneous rhythmic activity after application of neurotransmitter antagonists to spinal networks of the chick embryo. AB - We studied the regulation of spontaneous activity in the embryonic (day 10-11) chick spinal cord. After bath application of either an excitatory amino acid (AP 5 or CNQX) and a nicotinic cholinergic (DHbetaE or mecamylamine) antagonist, or glycine and GABA receptor (bicuculline, 2-hydroxysaclofen, and strychnine) antagonists, spontaneous activity was blocked for a period (30-90 min) but then reappeared in the presence of the drugs. The efficacy of the antagonists was assessed by their continued ability to block spinal reflex pathways during the reappearance of spontaneous activity. Spontaneous activity ceased over the 4-5 hour monitoring period when both sets of antagonists were applied together. After application of glycine and GABA receptor antagonists, the frequency of occurrence of spontaneous episodes slowed and became highly variable. By contrast, during glutamatergic and nicotinic cholinergic blockade, the frequency of occurrence of spontaneous episodes initially slowed and then recovered to stabilize near the predrug level of activity. Whole-cell recordings made from ventral spinal neurons revealed that this recovery was accompanied by an increase in the amplitude of spontaneously occurring synaptic events. We also measured changes in the apparent equilibrium potential of the rhythmic, synaptic drive of ventral spinal neurons using voltage or discontinuous current clamp. After excitatory blockade, the apparent equilibrium potential of the rhythmic synaptic drive shifted approximately 10 mV more negative to approximately -30 mV. In the presence of bicuculline, the apparent equilibrium potential of the synaptic drive shifted toward the glutamate equilibrium potential. Considered with other evidence, these findings suggest that spontaneous rhythmic output is a general property of developing spinal networks, and that GABA and glycinergic networks alter their function to compensate for the blockade of excitatory transmission. PMID- 9412509 TI - Platelet-activating factor receptor stimulation disrupts neuronal migration In vitro. AB - LIS-1 is a gene whose hemi-deletion causes the human neuronal migration disorder Miller-Dieker lissencephaly. It encodes a subunit of a brain platelet-activating factor (PAF) acetylhydrolase, an enzyme that inactivates PAF by hydrolyzing the acetyl moiety in the sn2 position of this phospholipid. Because PAF receptor activation has been shown to affect the developing neuronal cytoskeleton, we have hypothesized that a role for PAF in neurodevelopment is that of a modulator of neuroblast movement (a cytoskeletal function) and that an aberrant regulation of PAF could lead to an early arrest in migration. This report examines the effects of the nonhydrolyzable PAF receptor agonist methyl carbamyl PAF (mc-PAF) on the unidirectional in vitro migration of granule cells from cerebellar cell reaggregates on a laminin substrate. Bath treatment with mc-PAF yields a dose dependent decrease in granule cell migration compared with controls. This effect can be blocked by the simultaneous bath application of BN 52021 and trans-BTD, PAF receptor-specific antagonists. Although mc-PAF minimally inhibited neurite growth, its primary effect was on somal movement along preextended neurites. These experiments suggest that the stimulation of neuronal PAF receptors could be one crucial step for the regulation of neuroblast migration and that disturbed PAF catabolism during neurodevelopment could contribute to the neuronal migration defects observed in Miller-Dieker lissencephaly. PMID- 9412510 TI - Postnatal expression of Hu-bcl-2 gene in Lurcher mutant mice fails to rescue Purkinje cells but protects inferior olivary neurons from target-related cell death. AB - The Lurcher mutant has been extensively studied as a model for cell-autonomous and target-related cell death, yet there are still many unknowns concerning the mechanisms of neuronal degeneration in this mutant. As a key regulator of apoptosis, a bcl-2 transgene has been overexpressed in the heterozygous Lurcher mutant to investigate the effects of BCL-2 on two types of in vivo neuronal cell loss in Lurcher: cell-autonomous Purkinje cell degeneration and target-related olivary neuron death. Six adult +/Lc mutants expressing a human bcl-2 transgene (Hu-bcl-2) were generated by crossing +/Lc mutants with NSE71 Hu-bcl-2 transgenic mice. Analysis of these brains showed that bcl-2 overexpression did not prevent +/Lc Purkinje cell degeneration, but it did rescue most olivary neurons from target-related cell death. Although the number of olivary neurons was equivalent to wild-type numbers, the inferior olive nucleus was significantly shorter in its rostrocaudal extent, suggesting that olivary neurons are atrophied. We propose that Lurcher gene action causes Purkinje cell degeneration independently of a BCL 2-mediated pathway. Furthermore, although bcl-2 overexpression rescues olivary neurons from target-related cell death, it does not prevent the atrophy associated with the loss of target-related trophic support. PMID- 9412511 TI - Comparison of neurite outgrowth induced by intact and injured sciatic nerves: a confocal and functional analysis. AB - Mechanisms regulating axon growth in the peripheral nervous system have been studied by means of an in vitro bioassay, the tissue section culture, in which regenerating neurons are grown on substrata made up of tissue sections. Sections from intact and degenerated sciatic nerves proved to be different in their ability to support neurite outgrowth of embryonic chick sensory neurons from both qualitative and quantitative points of view. On denervated nerve sections, the total length of neurites elaborated per neuron was almost twice that found on intact nerve sections. In addition, confocal microscopy revealed a striking difference between intact and denervated nerve substrata: on denervated nerve sections, neurites grew inside the internal structures of endoneurial Schwann cell tubes, within the underlying tissue sections, whereas on intact nerve sections neurites extended along endoneurial basal laminae but never entered Schwann cell tubes. Perturbation experiments were used to analyze some of the molecular determinants that control neurite outgrowth in this system. Antibodies directed against the beta1-integrin subunit inhibited neurite extension on both normal and degenerated rat sciatic nerve tissue. Strikingly, however, differential inhibition was observed using antibodies directed against extracellular matrix molecules. Anti-laminin-2 (merosin) antibodies drastically reduced both the percentage of growing neurons and the total length of neurites on denervated nerve sections, but they did not modify these parameters on sections of normal nerve. Taken together, these results suggest that laminin 2/merosin promotes neurite outgrowth in peripheral nerve environments but only after Wallerian degeneration, which is when axons are allowed to extend within endoneurial tubes. PMID- 9412512 TI - Development of walking, swimming and neuronal connections after complete spinal cord transection in the neonatal opossum, Monodelphis domestica. AB - Development of coordinated movements was quantitatively assessed in adult opossums (Monodelphis domestica) with thoracic spinal cords transected by (1) crushing 7-8 d after birth [postnatal days 7-8 (P7-P8)]; at 2-3 years of age, systematic behavioral tests (e.g., climbing, footprint analysis, and swimming) showed only minor differences between control (n = 5) and operated (n = 10) animals; and (2) cutting on P4-P6; at 1 month these opossums exhibited coordinated walking movements but were unable to right themselves from a supine position, unlike controls (n = 6). When tested at 2 or 6 months, they could right themselves and showed remarkable coordination, albeit with more differences from controls than after a crush. No animals with spinal cords that were crushed at P14-18 survived because of cannibalism by the mother. Morphological studies (n = 10) 3 months-3 years after crush at 1 week showed restoration of structural continuity and normal appearance at the lesion site. Animals with cut rather than crushed cords showed continuity but greater morphological deficits. That lesions were complete was demonstrated by examining morphology and nerve impulse conduction immediately after crushing or cutting the spinal cord in controls. After lumbar spinal cord injection of 10 kDa dextran amine, retrogradely labeled cells were found rostral to the lesion in hindbrain and midbrain nuclei. Conduction was restored across the site of the lesion. Thus complete spinal cord transection in neonatal Monodelphis was followed by development of coordinated movements and repair of the spinal cord, a process that included development of functional connections by axons that crossed the lesion. PMID- 9412513 TI - Peripheral target regulation of the development and survival of spinal sensory and motor neurons in the chick embryo. AB - Unilateral limb-bud removal (LBR) before the outgrowth of sensory or motor neurons to the leg of chick embryos was used to examine the role of limb (target) derived signals in the development and survival of lumbar motoneurons and sensory neurons in the dorsal root ganglia (DRG). After LBR, motor and sensory neurons underwent normal initial histological differentiation, and cell growth in both populations was unaffected. Before their death, target-deprived motoneurons also expressed a cell-specific marker, the homeodomain protein islet-1. Proliferation of sensory and motor precursor cells was also unaffected by LBR, and the migration of neural crest cells to the DRG and of motoneurons into the ventral horn occurred normally. During the normal period of programmed cell death (PCD), increased numbers of both sensory and motor neurons degenerated after LBR. However, whereas motoneuron loss increased by 40-50% (90% total), only approximately 25% more sensory neurons degenerated after LBR. A significant number of the surviving sensory neurons projected to aberrant targets in the tail after LBR, and many of these were lost after ablation of both the limb and tail. Treatment with neurotrophic factors (or muscle extract) rescued sensory and motor neurons from cell death after LBR without affecting precursor proliferation of either population. Activity blockade with curare failed to rescue motoneurons after LBR, and combined treatment with curare plus muscle extract was no more effective than muscle extract alone. Treatment with the antioxidant N acetylcysteine rescued motoneurons from normal cell death but not after LBR. Two specific inhibitors of the interleukin beta1 converting enzyme (ICE) family of cysteine proteases also failed to prevent motoneuron death after LBR. Taken together these data provide definitive evidence that the loss of spinal neurons after LBR cannot be attributed to altered proliferation, migration, or differentiation. Rather, in the absence of limb-derived trophic signals, the affected neurons fail to survive and undergo PCD. Although normal cell death and cell death after target deprivation share many features in common, the intracellular pathways of cell death in the two may be distinct. PMID- 9412514 TI - Organization and transmitter specificity of medullary neurons activated by sustained hypertension: implications for understanding baroreceptor reflex circuitry. AB - In situ expression of c-fos observed in response to phenylephrine (PE)-induced hypertension provided a basis for characterizing the organization and neurotransmitter specificity of neurons at nodal points of medullary baroreflex circuitry. Sustained hypertension induced by a moderate dose of PE provoked patterns of c-fos mRNA and protein expression that conformed in the nucleus of the solitary tract (NTS) to the termination patterns of primary baroreceptor afferents and in the caudal ventrolateral medulla (CVLM) to a physiologically defined depressor region. A majority of barosensitive CVLM neurons concurrently displayed markers for the GABAergic phenotype; few were glycinergic. Phenylephrine-sensitive GABAergic neurons that were retrogradely labeled after tracer deposits in pressor sites of the rostral ventrolateral medulla (RVLM) occupied a zone extending approximately 1.4 mm rostrally from the level of the calamus scriptorius, intermingled partly with catecholaminergic neurons of the A1 and C1 cell groups. By contrast, barosensitive neurons of the NTS were found to be phenotypically complex, with very few projecting directly to the RVLM. Extensive colocalization of PE-induced Fos-IR and markers for the nitric oxide phenotype were seen in a circumscribed, rostral, portion of the baroreceptor afferent zone of the NTS, whereas only a small proportion of PE-sensitive neurons in the NTS were found to be GABAergic. PE treatment parameters have been identified that provide a basis for defining and characterizing populations of neurons at the first station in the central processing of primary baroreceptor input and at a key inhibitory relay in the CVLM. PMID- 9412515 TI - Gamma oscillations in the entorhinal cortex of the freely behaving rat. AB - Gamma frequency field oscillations (40-100 Hz) are nested within theta oscillations in the dentate-hilar and CA1-CA3 regions of the hippocampus during exploratory behaviors. These oscillations reflect synchronized synaptic potentials that entrain the discharge of neuronal populations within the approximately 10-25 msec range. Using multisite recordings in freely behaving rats, we examined gamma oscillations within the superficial layers (I-III) of the entorhinal cortex. These oscillations increased in amplitude and regularity in association with entorhinal theta waves. Gamma waves showed an amplitude minimum and reversed in phase near the perisomatic region of layer II, indicating that they represent synchronized synaptic potentials impinging on layer II-III neurons. Theta and gamma oscillations in the entorhinal cortex were coupled with theta and gamma oscillations in the dentate hilar region. The majority of layer II-III neurons discharged irregularly but were phase-related to the negative peak of the local (layer II-III) gamma field oscillation. These findings demonstrate that layer II-III neurons discharge in temporally defined gamma windows (approximately 10-25 msec) coupled to the theta cycle. This transient temporal framework, which emerges in both the entorhinal cortex and the hippocampus, may allow spatially distributed subpopulations to form temporally defined ensembles. We speculate that the theta-gamma pattern in the discharge of these neurons is essential for effective neuronal communication and synaptic plasticity in the perforant pathway. PMID- 9412516 TI - A model that accounts for activity in primate frontal cortex during a delayed matching-to-sample task. AB - A fully recurrent neural network model was optimized to perform a spatial delayed matching-to-sample task (DMS). In DMS, a stimulus is presented at a sample location, and a match is reported when a subsequent stimulus appears at that location. Stimuli elsewhere are ignored. Computationally, a DMS system could consist of memory and comparison components. The model, although not constrained to do so, worked by using two corresponding classes of neurons in the hidden layer: storage and comparator units. Storage units form a dynamical system with one fixed point attractor for each sample location. Comparator units constitute a system receiving input from these storage units as well as from current input stimuli. Both unit types were tuned directionally. These two sources of information combine to create unique patterns of activity that determine whether a match has occurred. In networks with abundant hidden units, the storage and comparator functions were distributed so that individual units took part in both. We compared the model with single-neuron recordings from premotor (PM) and prefrontal (PF) cortex. As shown previously, many PM and PF neurons behaved like storage units. In addition, both regions contain neurons that behave like the comparator units of the model and appear to have dual functionality similar to that observed in the model units. No neuron in either area had properties identical to those of the match output neuron of the model. However, four PF neurons and one PM neuron resembled the output signal more closely than any of the hidden units of the model. PMID- 9412517 TI - Masked presentations of emotional facial expressions modulate amygdala activity without explicit knowledge. AB - Functional magnetic resonance imaging (fMRI) of the human brain was used to study whether the amygdala is activated in response to emotional stimuli, even in the absence of explicit knowledge that such stimuli were presented. Pictures of human faces bearing fearful or happy expressions were presented to 10 normal, healthy subjects by using a backward masking procedure that resulted in 8 of 10 subjects reporting that they had not seen these facial expressions. The backward masking procedure consisted of 33 msec presentations of fearful or happy facial expressions, their offset coincident with the onset of 167 msec presentations of neutral facial expressions. Although subjects reported seeing only neutral faces, blood oxygen level-dependent (BOLD) fMRI signal in the amygdala was significantly higher during viewing of masked fearful faces than during the viewing of masked happy faces. This difference was composed of significant signal increases in the amygdala to masked fearful faces as well as significant signal decreases to masked happy faces, consistent with the notion that the level of amygdala activation is affected differentially by the emotional valence of external stimuli. In addition, these facial expressions activated the sublenticular substantia innominata (SI), where signal increases were observed to both fearful and happy faces--suggesting a spatial dissociation of territories that respond to emotional valence versus salience or arousal value. This study, using fMRI in conjunction with masked stimulus presentations, represents an initial step toward determining the role of the amygdala in nonconscious processing. PMID- 9412518 TI - Brain dopamine neurotoxicity in baboons treated with doses of methamphetamine comparable to those recreationally abused by humans: evidence from [11C]WIN 35,428 positron emission tomography studies and direct in vitro determinations. AB - The present study sought to determine whether doses of methamphetamine in the range of those used recreationally by humans produce brain dopamine (DA) neurotoxicity in baboons and to ascertain whether positron emission tomography (PET) imaging with the DA transporter (DAT) ligand [11C]WIN-35,428 ([11C]2beta carbomethoxy-3beta-(4-fluorophenyl)-tropane) could be used to detect methamphetamine-induced DAT loss in living primates. Baboons were treated with saline (n = 3) or one of three doses of methamphetamine [0.5 mg/kg (n = 2); 1 mg/kg (n = 2); and 2 mg/kg (n = 3)], each of which was given intramuscularly four times at 2 hr intervals. PET studies were performed before and 2-3 weeks after methamphetamine treatment. After the final PET studies, animals were killed for direct neurochemical determination of brain DA axonal markers. PET-derived binding potential values, used to index striatal DAT density, were significantly decreased after methamphetamine, with larger decreases occurring after higher methamphetamine doses. Reductions in striatal DAT documented by PET were associated with decreases in DA, dihydroxyphenylacetic acid, and specific [3H]WIN 35,428 and [3H]DTBZ binding determined in vitro. Decreases in DAT detected with PET were highly correlated with decreases in specific [3H]WIN-35,428 binding determined in vitro in the caudate of the same animal (r = 0.77; p = 0.042). These results indicate that methamphetamine, at doses used by some humans, produces long-term reductions in brain DA axonal markers in baboons, and that it is possible to detect methamphetamine-induced DAT loss in living nonhuman primates by means of PET. PMID- 9412519 TI - Dissociation Of working memory from decision making within the human prefrontal cortex. AB - We tested the hypothesis that cognitive functions related to working memory (assessed with delay tasks) are distinct from those related to decision making (assessed with a gambling task), and that working memory and decision making depend in part on separate anatomical substrates. Normal controls (n = 21), subjects with lesions in the ventromedial (VM) (n = 9) or dorsolateral/high mesial (DL/M) prefrontal cortices (n = 10), performed on (1) modified delay tasks that assess working memory and (2) a gambling task designed to measure decision making. VM subjects with more anterior lesions (n = 4) performed defectively on the gambling but not the delay task. VM subjects with more posterior lesions (n = 5) were impaired on both tasks. Right DL/M subjects were impaired on the delay task but not the gambling task. Left DL/M subjects were not impaired on either task. The findings reveal a cognitive and anatomic double dissociation between deficits in decision making (anterior VM) and working memory (right DL/M). This presents the first direct evidence of such effects in humans using the lesion method and underscores the special importance of the VM prefrontal region in decision making, independent of a direct role in working memory. PMID- 9412520 TI - Spatial distribution of potentiated synapses in hippocampus: dependence on cellular mechanisms and network properties. AB - Long-term potentiation (LTP) of synaptic transmission, studied intensively in reduced brain preparations such as hippocampal brain slices, is the leading candidate for the cellular/molecular basis of learning and memory. Serious consideration of LTP as underlying information storage in the intact brain, however, requires understanding how LTP can be induced selectively at specific synaptic sites in a neural system when the mechanisms underlying LTP are regulated by other structural and functional properties of the same neural system. In the studies reported here, we tested the hypothesis that different patterns of activity within the same population of entorhinal cortical afferents could lead to a selective potentiation of spatially distinct populations of synapses across different regions of the hippocampus, including those activated multisynaptically. We focused specifically on potentiation of direct, monosynaptic entorhinal input to dentate granule cells, which expresses an NMDA receptor-dependent LTP, and on potentiation of indirect, disynaptic entorhinal input to CA3 pyramidal cells, which is transmitted by the mossy fiber projection of dentate granule cells and expresses an NMDA receptor-independent LTP. The principal findings of these experiments show that lower stimulation frequencies (10-20 Hz) of entorhinal cortical axons selectively induce LTP of mossy fiber input to CA3 transsynaptically via excitation of dentate granule cells, and that patterns of stimulation of that mimic neuronal firing in the entorhinal cortex during endogenous theta rhythm (five-impulse bursts at 200 Hz, interburst intervals of 200 msec) induce LTP both monosynaptically for input to dentate granule cells and transsynaptically for mossy fiber input to CA3. PMID- 9412521 TI - Hypoactivity of the spinal cannabinoid system results in NMDA-dependent hyperalgesia. AB - Cannabinoids, such as Delta9-THC, are capable of inhibiting nociception, i.e., pain transmission, at least in part, by interacting with spinal Gi/Go-coupled cannabinoid receptors. What is not known, however, is the antinociceptive role of endogenous spinal cannabinoids. If endogenous cannabinoids modulate basal nociceptive thresholds, then alterations in this system could be involved in the etiology of certain pain states. In this report we provide evidence for tonic modulation of basal thermal nociceptive thresholds by the spinal cannabinoid system. Administration of oligonucleotides directed against CB1 cannabinoid receptor mRNA significantly reduced spinal cannabinoid binding sites and produced significant hyperalgesia when compared with a randomer oligonucleotide control. A second method used to reduce activity of the spinal cannabinoid receptor was intrathecal administration of the cannabinoid receptor antagonist SR 141716A. SR 141716A evoked thermal hyperalgesia with an ED50 of 0.0012 fmol. The SR 141716A induced hyperalgesia was dose-dependently blocked by the administration of D-AP-5 or MK-801, two antagonists to the NMDA receptor. These results indicate that there is tonic activation of the spinal cannabinoid system under normal conditions. Furthermore, hypoactivity of the spinal cannabinoid system results in an NMDA-dependent hyperalgesia and thus may participate in the etiology of certain chronic pain states. PMID- 9412522 TI - Involvement of presynaptic and postsynaptic mechanisms in a cellular analog of classical conditioning at Aplysia sensory-motor neuron synapses in isolated cell culture. AB - Temporal pairing of presynaptic activity and serotonin produces enhanced facilitation at Aplysia sensory-motor neuron synapses (pairing-specific facilitation), which may contribute to classical conditioning of the gill and siphon withdrawal reflex. This cellular analog of conditioning is thought to involve Ca2+ priming of the cAMP pathway in the sensory neurons. Consistent with that idea, we have found that pairing-specific facilitation by serotonin is greatly reduced by presynaptic injection of a slow Ca2+ chelator or a specific inhibitor of cAMP-dependent protein kinase and is accompanied by a transient increase in the frequency but by no change in the amplitude of spontaneous, miniature EPSPs. However, like post-tetanic potentiation (PTP) and long-term potentiation (LTP) at these synapses, pairing-specific facilitation is also greatly reduced by postsynaptic injection of a rapid Ca2+ chelator or by postsynaptic hyperpolarization during training, although postsynaptic hyperpolarization has no effect on the increase in frequency or on the amplitude of spontaneous EPSPs. These results suggest that pairing-specific facilitation by serotonin involves Hebbian postsynaptic as well as non-Hebbian presynaptic components that interact in some way, perhaps via retrograde signaling, to specifically enhance evoked, synchronized release of transmitter. PMID- 9412523 TI - Reconstruction of flexor/extensor alternation during fictive rostral scratching by two-site stimulation in the spinal turtle with a transverse spinal hemisection. AB - Analyses of fictive scratching motor patterns in the spinal turtle with transverse hemisection provided support for the concept of bilateral shared spinal cord circuitry among neurons responsible for generating left- and right side rostral, pocket, and caudal fictive scratching. Rhythmic bursts of hip flexor activity, the hip extensor deletion variation of fictive rostral scratching, were elicited by ipsilateral stimulation in the rostral scratch receptive field of a spinal turtle [transection at the segmental border between the second (D2) and third (D3) postcervical spinal segments] with a contralateral transverse hemisection one segment anterior to the hindlimb enlargement (at the D6-D7 segmental border). In addition, other sites were stimulated in this preparation: (1) contralateral sites in a rostral, pocket, or caudal scratch receptive field or (2) ipsilateral sites in a caudal scratch receptive field. A reconstructed fictive rostral scratch motor pattern of rhythmic alternation between hip flexor and hip extensor activation was produced by simultaneous stimulation of one site in the ipsilateral rostral scratch receptive field and another site in one of the other scratch receptive fields. This reconstructed rostral scratch motor pattern resembled the normal rostral scratch motor pattern produced by one-site rostral scratch stimulation of a spinal turtle (D2-D3 transection) with no additional transections. The observation of a reconstructed rostral scratch motor pattern produced by two-site stimulation in the spinal turtle with transverse hemisection supports the concept that hip extensor circuitry activated by stimulation of other scratch receptive fields is shared with circuitry activated by ipsilateral rostral scratch receptive field stimulation. PMID- 9412524 TI - Differential modulation of changes in hippocampal-septal synaptic excitability by the amygdala as a function of either elemental or contextual fear conditioning in mice. AB - Recent data obtained using a classic fear conditioning paradigm showed a dissociation between the retention of associations relative to contextual information (dependent on the hippocampal formation) and the retention of elemental associations (dependent on the amygdala). Furthermore, it was reported that conditioned emotional responses (CERs) could be dissociated from the recollection of the learning experience (declarative memory) in humans and from modifications of the hippocampal-septal excitability in animals. Our aim was to determine whether these two systems ("behavioral expression" system and "factual memory" system) interact by examining the consequences of amygdalar lesions (1) on the modifications of hippocampal-septal excitability and (2) on the behavioral expression of fear (freezing) resulting from an aversive conditioning during reexposure to conditional stimuli (CSs). During conditioning, to modulate the predictive nature of the context and of a discrete stimulus (tone) on the unconditional stimulus (US) occurrence, the phasic discrete CS was paired with the US or randomly distributed with regard to the US. After the lesion, the CER was dramatically reduced during reexposure to the CSs, whatever the type of acquisition. However, the changes in hippocampal-septal excitability persisted but were altered. For controls, a decrease in septal excitability was observed during reexposure to the conditioning context only for the "unpaired group" (predictive context case). Conversely, among lesioned subjects this decrease was observed in the "paired group" (predictive discrete CS case), whereas this decrease was significantly reduced in the unpaired group with respect to the matched control group. The amplitude and the direction of these modifications suggest a differential modulation of hippocampal-septal excitability by the amygdala to amplify the contribution of the more predictive association signaling the occurrence of the aversive event. PMID- 9412525 TI - Whole-cell plasticity in cocaine withdrawal: reduced sodium currents in nucleus accumbens neurons. AB - The nucleus accumbens is a forebrain region that mediates cocaine self administration and withdrawal effects in animal models of cocaine dependence. Considerable evidence suggests an important role of dopamine D1 receptors in these effects. Using a combination of current-clamp recordings in brain slices and whole-cell patch-clamp recordings from freshly dissociated neurons, we found that nucleus accumbens neurons are less excitable in cocaine withdrawn rats because of a novel form of plasticity: reduced whole-cell sodium currents. Three days after discontinuation of repeated cocaine injections, nucleus accumbens neurons recorded in brain slices were less responsive to depolarizing current injections, had higher action potential thresholds, and had lower spike amplitudes. Freshly dissociated nucleus accumbens neurons from cocaine-pretreated rats exhibited diminished sodium current density and a depolarizing shift in the voltage-dependence of sodium channel activation. These effects appear to be related to enhanced basal phosphorylation of sodium channels because of increased transmission through the dopamine D1 receptor/cAMP-dependent protein kinase pathway. The effects of repeated cocaine administration were not mimicked by repeated injections of the local anesthetic lidocaine and were not observed in neurons within the motor cortex, indicating that they did not result from local anesthetic actions of cocaine. Because nucleus accumbens neurons are normally recruited to coordinate response patterns of movement and affect, the decreased excitability during cocaine withdrawal may be related to symptoms such as anergia, anhedonia, and depression. PMID- 9412526 TI - Conversion of sensory signals into motor commands in primary motor cortex. AB - Movement triggered by sensory stimuli requires that the networks generating the motor commands receive an adequate driving input, which, in general, is a transformed version of the initial sensory signal. We investigated the nature of this transformation in a task in which monkeys categorize the speed of tactile stimuli as either low or high, reaching for one of two pushbuttons to indicate their choice. Extracellular recordings from primary motor cortex revealed two types of neurons selective for the speed categories: ones that fire at higher rates for low versus high speeds, and others that do the opposite. These differential responses are task-specific; no firing rate modulation was seen when identical arm movements were triggered by visual cues or when stimuli were delivered passively. Analyses using decoding and modeling techniques produced two main results. First, the neurons accurately encode the chosen category; an observer measuring their responses can exhibit a psychophysical performance during categorization identical to the monkey's. Second, by analyzing separately the trials in which hits and errors were scored, it is possible to distinguish purely sensory activity from activity exclusively related to arm motion. The recorded responses did not match either of these alternatives but were consistent with a model in which the category-tuned neurons are the link between the output of the sensory categorization process and the motor command used to indicate the animal's decision. Thus, the observed activity seems to encode a preprocessed version of the sensory stimulus and to participate in driving the arm motion. PMID- 9412527 TI - Postnatal development of phase-locked high-fidelity synaptic transmission in the medial nucleus of the trapezoid body of the rat. AB - Synaptic transmission in the medial nucleus of the trapezoid body of rats was analyzed in postnatal days 4-13 (P4-P13) by applying the whole-cell patch recording technique to brain slices. In P4-P6 animals, evoked EPSCs fluctuated extensively in amplitude and occurred in marked asynchrony, followed by spontaneous EPSCs. With development of animals, the evoked EPSCs increased in amplitude, and the rise time became faster. In addition, the synaptic transmission became phase-locked. The coefficient of variation (CV) of EPSC amplitude decreased with development (0.32 +/- 0.03 for P4-P5 and 0. 05 +/- 0.01 for P9-P11). The amplitude of miniature EPSCs did not change throughout the postnatal days investigated (-30.2 +/- 0.3 pA at -70 mV). The CV was dependent on extracellular Ca2+ concentration ([Ca2+]o) and was reduced with the increase of [Ca2+]o, and this [Ca2+]o dependence was shifted toward lower [Ca2+]o with development. Direct patch recording of the presynaptic terminals demonstrated an increase in Ca2+ currents during these postnatal days. The phase-locked high fidelity transmission in this synapse is achieved with development likely through the increase of Ca2+ currents and Ca2+ sensitivity of transmitter release mechanisms in the presynaptic terminal. PMID- 9412528 TI - Peripheral neural mechanisms determining the orientation of cylinders grasped by the digits. AB - When a human grasps a cylindrical object, feedback on the orientation of the cylinder with respect to the axes of the digits is crucial for successful manipulation of the object. We measured the ability of humans to discriminate the orientations of cylinders passively contacting the fingerpad. For a cylinder of curvature of 521 m-1 (radius, 1.92 mm) subjects were able to discriminate, at the 75% level, orientation differences of 5.4 degrees; for a less curved cylinder (curvature, 172 m-1; radius, 5.81 mm) the difference limen decreased to 4.2 degrees. The neural mechanisms underlying the determination of tactile orientation were investigated by recording the responses of single slowly adapting type I afferents (SAIs) innervating the fingerpads of anesthetized monkeys. When cylinders were stepped across the receptive field of an SAI, the resulting response profiles were Gaussian in shape; the shape corresponded to the shape of the cylinder, increasing in height and decreasing in width for more curved cylinders. All SAIs had the same underlying profile shape except for a multiplicative constant determined by the sensitivity of the individual afferent. Thus it was possible to reconstruct the response of the population of active SAIs in the fingerpad. Changing the orientation of the cylinder resulted in a rotation of the population response, but the change in angle of the population response was greater than the change in orientation of the cylinder. This discrepancy increased as the orientation of the cylinder moved closer to the orientation of the axis of the finger and was more pronounced for the less curved cylinder. Measured contact areas between the cylinders and the skin were elliptical, with orientations exceeding those of the cylinder; again the differences were greater for the less curved cylinder and for orientations closer to that of the finger axis. The human discrimination performance could be explained in terms of the SAI population responses. PMID- 9412529 TI - A model for encoding multiple object motions and self-motion in area MST of primate visual cortex. AB - Many cells in the dorsal part of the medial superior temporal (MST) region of visual cortex respond selectively to specific combinations of expansion/contraction, translation, and rotation motions. Previous investigators have suggested that these cells may respond selectively to the flow fields generated by self-motion of an observer. These patterns can also be generated by the relative motion between an observer and a particular object. We explored a neurally constrained model based on the hypothesis that neurons in MST partially segment the motion fields generated by several independently moving objects. Inputs to the model were generated from sequences of ray-traced images that simulated realistic motion situations, combining observer motion, eye movements, and independent object motions. The input representation was based on the response properties of neurons in the middle temporal area (MT), which provides the primary input to area MST. After applying an unsupervised optimization technique, the units became tuned to patterns signaling coherent motion, matching many of the known properties of MST cells. The results of this model are consistent with recent studies indicating that MST cells primarily encode information concerning the relative three-dimensional motion between objects and the observer. PMID- 9412530 TI - A focal zone of thalamic plasticity. AB - In this study, sensory maps in the thalamus were investigated by examining their volume and shape. We determined the forelimb representation in adult rats after the removal of hindlimb input by nucleus gracilis lesions. Three-dimensional reconstructions of thalamic sensory maps were obtained from a grid of electrode penetrations. We found that the volume of the shoulder sensory map contracted >50% at an acute time interval (n = 6), followed by a robust volumetric sensory map expansion of 25% at 1 week (n = 8) and 1 month (n = 8) after lesion relative to controls (n = 8). The topology of the volumetric increase was scrutinized by slicing functional maps in the coronal, sagittal, and horizontal planes. The equivalence of such slices from each animal was established by virtue of their distance from either a functional or neuroanatomical landmark. Surprisingly, all of the volumetric increase unequivocally occurred in a circumscribed coronal slice 300 micron thick. This focal zone was located toward the rostral pole of the thalamic tactile relay, the ventroposterolateral nucleus. Analysis in the sagittal plane revealed that, unexpectedly, the shoulder map volume expanded by superimposing its representation on that of the forepaw, via an advancement of the shoulder representation by 0.6 mm medially. We propose a "hot spot" hypothesis in which focal zones of plasticity may not be specific to the thalamus but may have manifestations elsewhere in the nervous system, such as the cerebral cortex or dorsal column nuclei. PMID- 9412531 TI - Leptin receptor immunoreactivity in chemically defined target neurons of the hypothalamus. AB - The adipose tissue-derived hormone leptin regulates body weight homeostasis by decreasing food intake and increasing energy expenditure. The weight-reducing action of leptin is thought to be mediated primarily by signal transduction through the leptin receptor (LR) in the hypothalamus. We have used immunohistochemistry to localize LR-immunoreactive (LR-IR) cells in the rat brain using an antiserum against a portion of the intracellular domain of LR that is common to all LR isoforms. The antiserum recognized the short and long isoforms of LR in transfected hematopoietic BaF3 cells. To examine the chemical nature of target cells for leptin, direct double-labeling immunofluorescence histochemistry was applied. The results show extensive distribution of LR-like immunoreactivity (LR-LI) in the brain with positively stained cells present, e.g., in the choroid plexus, cerebral cortex, hippocampus, thalamus, and hypothalamus. In the hypothalamus, strongly LR-IR neurons were present in the supraoptic nucleus (SON) and paraventricular nucleus (PVN), periventricular nucleus, arcuate nucleus, and lateral hypothalamus. Weaker LR-IR neurons were also demonstrated in the lateral and medial preoptic nuclei, suprachiasmatic nucleus, ventromedial and dorsomedial nuclei, and tuberomammillary nucleus. Confocal laser scanning microscopy showed LR-LI in the periphery of individual cells. In magnocellular neurons of the SON and PVN, LR-LI was demonstrated in vasopressin- and oxytocin-containing neurons. In parvocellular neurons of the PVN, LR-LI was demonstrated in many corticotropin releasing hormone-containing neurons. LR-IR neurons were mainly seen in the ventromedial aspect of the arcuate nucleus, where LR-LI co-localized with neuropeptide Y. In the ventrolateral part of the arcuate nucleus, LR-LI was present in many large adrenocorticotropic hormone-IR proopiomelanocortin containing neurons and in a few galanin-, neurotensin-, and growth hormone releasing hormone-containing neurons. In the dorsomedial arcuate nucleus, few tyrosine hydroxylase (dopamine)-containing neurons were seen to have LR-LI. Melanin-concentrating hormone-containing neurons in the lateral hypothalamus had LR-LI. Based on the immunohistochemical results, possible interactions of leptin with brain mechanisms are discussed. PMID- 9412532 TI - Directory of International Opportunities. 1997. PMID- 9412535 TI - 188th Meeting of the Society for Endocrinology. London, United Kingdom, 24-25 November 1997. Abstracts. PMID- 9412534 TI - Orthopaedic proceedings, 1996-1997. Abstracts. PMID- 9412536 TI - Pathological Society of Great Britain and Ireland 175th meeting. 2-4 July 1997. Abstracts. PMID- 9412537 TI - Overview of the treatment of heart failure. AB - Short-term goals of heart failure management are directed toward relieving symptoms such as shortness of breath, decreased exercise tolerance, and lower extremity edema and improving functional capacity and quality of life. Long-term goals include decreasing mortality and slowing or reversing the underlying cardiac structural abnormalities of heart failure. Improvement in symptomatic endpoints (e.g., exercise tolerance) does not necessarily correlate with endpoints for improved survival (e.g., left ventricular ejection fraction). It is therefore important to evaluate the effects of drugs on these distinct endpoints separately. Symptoms of heart failure are commonly managed with the use of diuretics, vasodilators, and positive inotropes or digoxin. Ideally, therapy should consist of a diuretic plus vasodilator (e.g., angiotensin-converting enzyme [ACE] inhibitor or isosorbide dinitrate plus hydralazine), with or without digoxin. Prevention of further left ventricular dysfunction can be accomplished by inhibiting neurohormonal processes and ventricular remodeling that occur in heart failure using ACE inhibitors, nitrates and hydralazine, or beta blockers. Significant therapeutic advances have been made with respect to symptom relief, hospitalizations, and mortality reduction in patients with congestive heart failure. Despite these advances, patient morbidity and mortality remain high and underscore the necessity for optimal use of existing therapies along with research directed at achieving further improvements in both quality of life and life expectancy. PMID- 9412538 TI - Adrenergic nervous system in heart failure. AB - Recent demonstration that the level of sympathetic nervous drive to the failing heart in patients with severe heart failure is a major determinant of prognosis, and that mortality in heart failure is decreased by beta-adrenergic blockade with carvedilol, indicates the clinical relevance of cardiac neuroscience research. Important initial findings were observations that the plasma concentration of the sympathetic neurotransmitter, norepinephrine, is elevated in heart failure, and that overall clinical outcome is related to plasma norepinephrine concentration (although heart failure severity may be a confounder). Sympathetic nerve recording (clinical microneurography) and radiotracer methods measuring regional sympathetic activity in the heart (cardiac norepinephrine "spillover") have now largely supplanted antecubital venous norepinephrine measurements as research tools, with newer methods providing information on regional sympathetic function that was previously lacking. The cardiac sympathetic nerves are preferentially stimulated in severe heart failure, with norepinephrine release from the failing heart at rest in untreated patients increased up to 50-fold, which is similar to the level of release in healthy hearts during near maximal exercise. There is lesser stimulation of the sympathetic outflows to the kidneys and skeletal muscle. In early mild heart failure, it is only the cardiac sympathetic nerves that are activated. This preferential activation of cardiac sympathetic outflow contributes to arrhythmia development and probably to progression of heart failure and has been linked to mortality in mild and severe heart failure. The central nervous system mechanisms involved in the sympathetic nervous activation present in patients with heart failure remain uncertain. Increased intracardiac diastolic pressure seems to be one peripheral reflex stimulus with increased forebrain norepinephrine turnover being an important central mechanism. PMID- 9412539 TI - Molecular and cellular mechanisms of myocardial failure. AB - Myocardial remodeling is a central feature in the progression of myocardial failure. This process, which can be stimulated by factors that are increased as a result of myocardial dysfunction such as mechanical stress, angiotensin, and norepinephrine, consists of a variety of molecular and cellular events that can lead to important changes in myocardial structure and function (or phenotype). These alterations include hypertrophy and cellular apoptosis of myocytes, changes in the molecular phenotype of the myocardium with reinduction of a fetal gene program, and alterations in the quantity and composition of the extracellular matrix. Agents that counteract these factors, such as vasodilators, angiotensin converting enzyme inhibitors, and beta-adrenergic antagonists, slow the progression of myocardial failure and are of clinical value in the treatment of heart failure. Several additional mechanisms have recently been identified that could also be important in mediating myocardial remodeling. These include oxidative stress, inflammatory cytokines, nitric oxide, endothelin, and peptide growth factors. It is likely that additional strategies to inhibit these mechanisms will exert beneficial effects on the process of myocardial remodeling and the development of clinical heart failure. PMID- 9412540 TI - Mechanism of action of beta-blocking agents in heart failure. AB - Antiadrenergic treatment is currently an emerging and very promising approach to the treatment of chronic heart failure. Although the adrenergic nervous system can be pharmacologically inhibited at multiple levels, it is the use of receptor blocking agents that has generated the most interest and provided the most data for the "proof of concept" of this approach. In part because antiadrenergic treatment of chronic heart failure has developed in an atmosphere in which it was initially considered to be contraindicated (i.e., before Phase III clinical trials could be initiated), a large body of hypothesis-driven basic and clinical investigation was required to define the overall rationale and demonstrate feasibility. This article will review these data and propose a single primary mechanism of action to explain most of the clinical benefits of these agents. PMID- 9412541 TI - Protective effects of carvedilol in the myocardium. AB - Beta blockers have long been used in the treatment of systemic hypertension, where they effectively lower blood pressure and, in so doing, they decrease left ventricular hypertrophy. The sympathetic nervous system is activated in patients with congestive heart failure, and therefore it is logical that beta blockers may also provide benefit in these patients. As such, beta blockers are currently being evaluated in several large clinical trials in congestive heart failure. One particular drug, carvedilol, is a third-generation vasodilating beta blocker that is marketed for the treatment of hypertension. The drug lowers systemic arterial blood pressure without producing reflex tachycardia and preserves renal function. Carvedilol decreases mortality by 65% and decreases hospitalization by 29% in patients with congestive heart failure. The effects of carvedilol in heart failure may result, at least in part, from beta blockade as well as vasodilation, the latter resulting from alpha(1)-adrenoceptor blockade. Interestingly, carvedilol has a number of additional properties that may also provide benefit in these patients. Carvedilol and several of its metabolites are potent antioxidants that may inhibit catecholamine toxicity resulting from the oxidation of norepinephrine and the subsequent formation of toxic intermediates, including the generation of reactive oxygen free radicals in the myocardium. As a result of its antioxidant activity, carvedilol also blocks the expression of several genes involved in myocardial damage and cardiac remodeling, and the drug inhibits free radical-induced activation of transcription factors and programmed cell death (apoptosis). Carvedilol is a novel beta blocker that is highly effective in the treatment of hypertension and congestive heart failure, and combines in one molecule a number of important pharmacologic properties. PMID- 9412542 TI - Effects of beta-adrenergic blockade on survival of patients with chronic heart failure. AB - Observations from experimental studies and controlled clinical trials indicate that prolonged activation of the sympathetic nervous system can accelerate the progression of heart failure, and that the risks of such progression can be substantially decreased through the use of pharmacologic agents that interfere with the actions of the sympathetic nervous system on the heart and peripheral blond vessels. Early studies with beta(1)-selective agents such as metoprolol and bisoprolol suggested that treatment with beta blockers could decrease the risk of worsening heart failure, but showed little or equivocal effects on survival. Recent studies using a more complex nonselective beta blocker (e.g., carvedilol) have reported a reduction in mortality as well as in the combined risk of death and hospitalization. This ability to decrease the risk of disease progression led to the recent approval of carvedilol for the treatment of chronic heart failure by the US Food and Drug Administration. However, it is not clear whether these survival effects represent a class effect of beta blockers or a specific effect of carvedilol. Carvedilol antagonizes several biologic mechanisms not blocked by metoprolol or bisoprolol that are thought to mediate the progression of heart failure. In addition, in 3 meta-analyses, the survival effects of nonselective vasodilating beta blockers appeared to be greater than those of beta(1)-selective nonvasodilating beta blockers. Moreover, it is unknown whether survival benefits can be expected only in the patients who were enrolled in the clinical trials (i.e., New York Heart Association class II or III patients) or whether they can also he achieved in patients with less severe or more severe symptoms (class I or IV heart failure). These questions are being addressed in several ongoing large scale, long-term survival trials that are scheduled for completion within the next 5 years. PMID- 9412543 TI - Effects of beta blockers on symptoms and functional capacity in heart failure. AB - Beta-adrenergic-blocking drugs ameliorate the progression of disease that usually characterizes heart failure. All of the larger multicenter studies have demonstrated reductions in morbidity, manifested by a reduction in the number of hospitalizations and/or listing for cardiac transplantation, whereas studies with carvedilol have also reported a significant reduction in mortality. The influence of beta-adrenergic blocking drugs on the symptomatic and functional status of patients with heart failure has been more difficult to establish. Almost all trials have reported an improvement in New York Heart Association (NYHA) functional class, with few patients experiencing functional deterioration. Improvement in symptom score was reported in the Metoprolol in Dilated Cardiomyopathy (MDC) trial far patients treated with metoprolol. In patients from the US Carvedilol Heart Failure Trials Program who had heart failure from ischemic and nonischemic etiologies, both patients and physicians reported more improvement and less deterioration in an assessment of global status. More formal instruments to assess quality of life, such as the specific activity scale in the Australia-New Zealand trial of carvedilol (that recruited 30% of patients who bad improved to NYHA stage I) and the Minnesota Quality of Life questionnaire, did not seem sensitive to the clinical benefits observed in these trials. Serial exercise testing similarly does not seem sensitive to the beneficial influence of beta-adrenergic blocking drugs on disease progression, at least in relatively short-term studies. PMID- 9412544 TI - OM-8BV for COPD. PMID- 9412545 TI - Making decisions about the forgoing of life-sustaining therapy. AB - The incidence of withholding and withdrawal of life support from critically ill patients has increased to the extent that these practices now precede well over half of all deaths in many intensive care units (ICUs). Although the forgoing of life-sustaining therapy is ethically acceptable and clinically desirable in certain instances, and although physicians do not have a responsibility to provide futile care even if a patient or surrogate insists on it, they must be cautious in exercising their influence, if not authority, over patients and surrogates in prompting the withholding and withdrawal of life support. Such caution is particularly indicated because managed care has made patients suspicious of healthcare institutions and physicians who are rewarded for restricting access to care. Most patients and surrogates agree with reasonable physician recommendations to forgo life-sustaining therapy. When they do not agree, physicians should not limit care on the basis of their own personal notions of futility, but should instead rely on institutional or multiinstitutional futility policies. Such policies should be developed by health professionals, patients, community leaders, and, when appropriate, participants in managed-care organizations. They should specify which ICU interventions are beneficial, address potential conflicts of interest, and be available to persons who could use such information in selecting the source of their care. PMID- 9412546 TI - Effects of an immunostimulating agent on acute exacerbations and hospitalizations in patients with chronic obstructive pulmonary disease. The PARI-IS Study Steering Committee and Research Group. Prevention of Acute Respiratory Infection by an Immunostimulant. AB - The PARI-IS Study is a double-blind placebo-controlled randomized clinical trial to study the effect of an immunostimulating agent to prevent acute respiratory exacerbation in patients with COPD. Three hundred eighty-one ambulatory patients (190 placebo and 191 immunostimulant) were followed at home for 6 mo by experienced research nurses. The risk of having at least one episode of acute exacerbation (primary outcome) was similar in the two groups (p = 0.872). In contrast, the total number of days of hospitalization for a respiratory problem was 55% less in the group treated with OM-85 BV (287 d) than in the group treated with placebo (642 d). Patients treated with OM-85 BV spent an average of 1.5 d in hospital compared with 3.4 d for patients treated with placebo (p = 0.037). The risk of being hospitalized for a respiratory problem was 30% lower in the treated group (16.2%) than in the placebo group (23.2%); p = 0.089. Eight deaths were observed: two in patients treated with OM-85 BV and six in patients treated with placebo (p = 0.153). During the course of the study dyspnea improved slightly in patients treated with OM-85 BV, whereas it deteriorated slightly in patients receiving placebo (p = 0.028). These results suggest that this immunostimulating agent may be beneficial for patients with COPD by reducing the likelihood of severe respiratory events leading to hospitalization. PMID- 9412547 TI - The leukotriene receptor antagonist zafirlukast inhibits sulfur dioxide-induced bronchoconstriction in patients with asthma. AB - Inhalation of sulfur dioxide (SO2) causes bronchoconstriction in most people with asthma. To examine the role of leukotrienes in this response, the antagonism of SO2-induced bronchoconstriction by a single oral dose of the leukotriene receptor antagonist zafirlukast was assessed in a double-blind, placebo-controlled, two period crossover trial in 12 subjects with mild-to-moderate asthma. Subjects had bronchial hyperresponsiveness, an FEV1 < or = 70% of predicted, and a positive response to inhaled SO2 (an 8-unit increase in specific airway resistance on inhaling an SO2 concentration of < or = 4 ppm (PC8SRaw). Subjects were treated with zafirlukast (20 mg) or placebo on two treatment days 5 to 14 d apart. Two and 10 hours after treatment, subjects inhaled SO2 (0.25, 0.5, 1.0, 2.0, 4.0, and 8.0 ppm) during eucapnic hyperventilation at 20 L/min. PC8SRaw was determined after each challenge. Blood samples were collected to assess zafirlukast plasma concentrations versus effect. PC8SRaw was significantly higher 2 h after zafirlukast compared with placebo (3.1 versus 1.5 ppm; p = 0.02) and remained higher 10 h after treatment with zafirlukast (2.7 versus 1.9 ppm; p = 0.09). An association was found between zafirlukast plasma concentrations and increases in PC8SRaw 10 h after treatment (p = 0.001). The safety profile of zafirlukast was not clinically different from placebo. A single 20-mg dose of zafirlukast attenuated SO2-induced bronchoconstriction. We conclude that S02-induced bronchoconstriction involves release of leukotrienes and that treatment with zafirlukast attenuates the bronchoconstrictor response. PMID- 9412548 TI - Comparison of salmeterol and albuterol-induced bronchoprotection against adenosine monophosphate and histamine in mild asthma. AB - Short-acting beta(2)-agonists provide greater protection to bronchoconstriction induced by adenosine-5'-monophosphate (AMP) than does methacholine. Because AMP produces bronchoconstriction through release of mediators from mast cells, and methacholine directly constricts airway smooth muscle, this suggests that beta(2) agonists stabilize mast cells in vivo. This in vivo property has not been demonstrated with long-acting beta(2)-agonists. We undertook two double-blind, randomized, crossover, placebo-controlled studies to investigate the effects of salmeterol and albuterol on airway responsiveness (AR) to AMP and histamine in patients with mild asthma. In the first study, 19 patients attended on four occasions to inhale salmeterol 50 micrograms or placebo 2 h before challenge with AMP or histamine. In the second study 16 patients (13 of whom had participated in the first study) were studied in a similar fashion but inhaled albuterol 400 micrograms or placebo 30 min prior to challenge. Salmeterol reduced AR to AMP and histamine by 3.4 +/- 0.3 and 3.9 +/- 0.3 doubling doses, respectively (NS). In contrast, albuterol demonstrated a greater protective effect on AMP than on histamine, reducing AR by 5.1 +/- 0.3 and 3.8 +/- 0.2 doubling doses, respectively (p < 0.005). Thus, in contrast to albuterol, salmeterol did not demonstrate mast-cell stabilizing properties in vivo at a time corresponding to maximal bronchodilatation. These findings might be explained by the unique pharmacologic profile of salmeterol in combination with the differential beta(2) adrenoceptor pharmacology of bronchial mast cells and bronchial smooth muscle. PMID- 9412549 TI - Effect of regular inhaled albuterol on allergen-induced late responses and sputum eosinophils in asthmatic subjects. AB - Treatment with inhaled beta(2)-agonists immediately before allergen inhalation inhibits allergen-induced early, but not late asthmatic responses (LAR). By contrast, 2 wk treatment with inhaled albuterol increases airway responses to inhaled allergen. We examined the effects of regular albuterol treatment on allergen-induced increases in inflammatory cells in blood and induced sputum. Ten mild, stable allergic asthmatics inhaled albuterol (800 micrograms/day) or placebo for 7 d in a controlled, randomized, double-blind, crossover study. Allergen inhalation was performed 12 h after the final dose. Methacholine airway responsiveness and blood samples were analyzed before and 24 h after, and induced sputum was obtained before, 7 h and 24 h after allergen. Allergen significantly reduced methacholine PC20, increased blood eosinophil numbers, and numbers of sputum neutrophils, EG2 positive and metachromatic cells (p < 0.05), without significant differences between treatments. Albuterol treatment significantly increased the LAR compared to placebo treatment (p = 0.003) and significantly enhanced the number of sputum eosinophils (p = 0.009) and sputum ECP (p = 0.04) at 7 h but not 24 h post-allergen (p > 0.05). We conclude that regular use of inhaled albuterol significantly increases the LAR to inhaled allergen, in association with an increase in the number of sputum eosinophils and the release of ECP, suggesting albuterol increases the late response by increasing eosinophil influx into the airways. PMID- 9412550 TI - Effects of budesonide and fluticasone on 24-hour plasma cortisol. A dose-response study. AB - Comparison of the risk-benefit profiles of different inhaled glucocorticoids has been limited by inadequate information about the dose-response relationships for efficacy relative to side effects. Fluticasone propionate (FP) is twice as effective as budesonide (BUD), but the potency ratio of FP:BUD with respect to suppression of cortisol production is unknown. The effects of 5 d of treatment with BUD (800, 1,600, and 3,200 micrograms/d via pMDI) and FP (750, 1,500, and 2,000 micrograms/d via pMDI) on integrated area under the curve of 24-h plasma cortisol profiles (AUC24 h) were compared in a randomized, placebo-controlled, seven-period crossover study in normal male volunteers (n = 28). Plasma cortisol concentrations were measured during the last 24 h of each treatment period. Each treatment (except BUD 800 micrograms) produced significant dose-dependent reductions in AUC24 h compared with placebo; e.g., percent reductions in AUC24 h were 23, 41, and 69% for the three doses of BUD, and, correspondingly, 46, 85, and 93% for the three doses of FP. Model-derived measurements of dose potency ratios showed that FP was 2.9 times more potent than BUD in reducing AUC24 h (95% CI, 2.5 to 3.5) and 3.1 times more potent in reducing 8:00 A.M. plasma cortisol (95% CI, 2.4 to 4.0). Thus, on a microgram-for-microgram notional dose basis, the systemic effects of a given dose of FP on AUC24 h cortisol were equivalent to the effects of three times the dose of BUD. PMID- 9412551 TI - Airway smooth muscle, tidal stretches, and dynamically determined contractile states. AB - In the classic theory of airway lumen narrowing in asthma, active force in airway smooth muscle is presumed to be in static mechanical equilibrium with the external load against which the muscle has shortened. This theory is useful because it identifies the static equilibrium length toward which activated airway smooth muscle would tend if given enough time. The corresponding state toward which myosin-actin interactions would tend is called the latch state. But are the concepts of a static mechanical equilibrium and the latch state applicable in the setting of tidal loading, as occurs during breathing? To address this question, we have studied isolated, maximally contracted bovine tracheal smooth muscle subjected to tidal stretches imposed at 0.33 Hz. We measured the active force (F) and stiffness (E), which reflect numbers of actin-myosin interactions, and hysteresivity (eta) which reflects the rate of turnover of those interactions. When the amplitude of imposed tidal stretch (epsilon) was very small, 0.25% of muscle optimal length, the steady-state value of F approximated the isometric force, E was large, and eta was small. When epsilon was increased beyond 1%, however, F and E promptly decreased and eta promptly increased. The muscle could be maintained in these steady, dynamically determined contractile states for as long as the tidal stretches were sustained; when epsilon subsequently decreased back to 0.25%, F, E, and eta returned slowly toward their previous values. The provocative stretch amplitude required to cause active force or muscle stiffness to fall by half, or hysteresivity to double, was slightly greater than 2%. These observations are consistent with a direct effect of stretch upon bridge dynamics in which, with increasing tidal stretch amplitude, the number of actin-myosin interactions decreases and their rate of turnover increases. We conclude that the interactions of myosin with actin are at every instant tending toward those that would prevail in the isometric steady state, but tidal changes of muscle length cause an excess in the rate of detachment. These stretch-induced detachment events can come so fast compared with the rate of attachment that static equilibrium conditions are never attained. If so, then airway lumenal narrowing and the underlying contractile state would be governed by a dynamic mechanical process rather than by a mechanical equilibrium of static forces. PMID- 9412552 TI - Sensitization to dust mites as a dominant risk factor for asthma among adolescents living in central Virginia. Multiple regression analysis of a population-based study. AB - Factors influencing asthma were investigated in a population of school children in central Virginia. A survey of 1,054 children in two middle schools (one urban and one suburban) identified 135 students with symptoms suggestive of asthma. Eighty-eight symptomatic children and 123 control subjects were randomly selected for further evaluation by skin testing using common indoor and outdoor allergens; serum assays for total IgE and specific IgE; dust samples assayed for mite (Der p 1 Der f 1), cat (Fel d 1), and cockroach (Bla g 2) allergens; and provocation with histamine to test for bronchial hyperreactivity. Forty-eight of the children with symptoms responded to < or = 3.9 mumol of histamine and were considered to have asthma. Marginal analysis identified elevated total IgE and dust mite, cat, and cockroach sensitization as significant risk factors for asthma. Using multiple regression, only dust mite sensitization was independently associated with asthma (odds ratio = 6.6; p < 0.0001). Dust from 81% of the houses contained high levels of mite allergen (> 2 micrograms/g), while approximately 40% of the children were exposed to cat and 17% were exposed to cockroach allergen. In this population, there was no significant association between asthma and race, socioeconomic status, home smoking, sensitization to outdoor allergens, or allergen concentration in the child's home. In an area where there is a high prevalence of asthma and most houses contain high concentrations of dust mite allergen, sensitization to this allergen is the dominant risk factor for asthma defined as symptomatic bronchial hyperreactivity PMID- 9412553 TI - Effects of photochemical air pollution and allergen exposure on upper respiratory tract inflammation in asthmatics. AB - Asthma is an inflammatory disease of the airways, and exacerbations of this disease have been associated with high levels of air pollution. The objective of this study was to examine whether ambient air pollution and/or allergen exposure induces inflammatory changes in the upper airways of asthmatics. Sixty patients with intermittent to severe persistent asthma visited the Hospital's Out Patient Clinic every 2 wk for a period of 3 mo, and on each visit a nasal lavage was obtained. Associations between nasal inflammatory parameters and seasonal allergens and/or air pollution exposures were analyzed using linear regression analysis. The study ran from July 3 to October 6, 1995, during which period ozone (8-h mean: 80 micrograms/m3) and PM10 (24-h mean: 40 micrograms/m3) were the major air pollutants; the major aeroallergen was mugwort pollen (24-h mean: 27 pollen grains/m3). Effects on both cellular and soluble markers in nasal lavage were demonstrated for both ozone and mugwort pollen, but not for PM10. Ambient ozone exposure was associated with an increase in neutrophils (112% per 100 micrograms/m3 increase in 8-h average ozone concentration), eosinophils (176%), epithelial cells (55%), IL-8 (22%), and eosinophil cationic protein (ECP) (19%). Increases in environmental mugwort pollen counts were associated with an increase in nasal eosinophils (107% per 100 pollen/m3) and ECP (23%), but not with neutrophils, epithelial cells, or lL-8. This study demonstrated that both ambient ozone and allergen exposure are associated with inflammatory responses in the upper airways of subjects with asthma, although the type of inflammation is qualitatively different. PMID- 9412554 TI - Genes for asthma? An analysis of the European Community Respiratory Health Survey. AB - Besides environmental triggers, a family history of asthma is a strong risk factor for the development of asthma in offspring. The pooled data from 13,963 interviews of randomly selected, 20 to 48 yr-old participants from the 30 centers of the European Community Respiratory Health Survey (ECRHS) were analyzed with conventional logistic regression and a Class A regressive model adapted for the segregation of various transmission modes in families. The asthma prevalence in the interviewed index generation was 6.9% (95% confidence interval [CI]: 6.5 to 7.3), and in the parent generation was 6.1% (5.8 to 6.4). As with asthma prevalence, the risk of a subject having asthma if a parent had asthma also had a large geographic variation across the survey centers. The mean risk if a father had asthma was 2.9 (2.4 to 3.5), and If the mother had asthma was 3.2 (2.6 to 3.9). The risk increased to 7.0 (3.9 to 12.7) if both parents were affected. For developing extrinsic asthma, extrinsic asthma in any parent was a greater risk factor (4.9 [3.9 to 6.0]) than intrinsic asthma of the parent (1.5 [0.8 to 2.6]), and the risk for women was slightly higher than that for men (4.3 [3.3 to 5.5] versus 3.6 [2.6 to 5.0]). Applying different segregation models, only a model for a two-allele gene with a codominant inheritance could not be rejected, assuming a major gene with a population frequency of 24.2%. Further results make a multilocus/threshold model likely. In conclusion, a history of asthma in parents is a strong risk factor for asthma in the offspring. Under the assumptions of the applied segregation analysis, at least one major gene exists which could be a gene involved also in allergy. However, asthma is not fully described by a single gene model. The risk for asthma varies within the European countries, and should be seen in the context of a complex genetic and environmental pathophysiology. PMID- 9412555 TI - A synoptic evaluation of asthma hospital admissions in New York City. AB - An evaluation of weather/asthma relationships in the New York City Standard Metropolitan Statistical Area (SMSA) is developed using a synoptic climatological methodology. This procedure isolates "air masses," or bodies of air that are homogeneous in meteorological character, and relates them to daily counts of overnight asthma hospital admissions. The synoptic procedure used here, known as the temporal synoptic index (TSI), can identify air masses in automated fashion for every day over many years. It is apparent that certain air masses are related to statistically significant increases in asthma hospital admissions. The impact varies seasonally, with weather having a particularly important impact on asthma admissions during fall and winter. It appears that air pollution has little impact on asthma during these two seasons, and the air masses associated with the highest admissions are not distinguished by high concentrations of pollutants. However, during spring and summer, the air masses associated with highest admissions are among those with high pollution concentrations. There is a strong interseasonal differential response to weather and air pollution by asthmatics in New York City. If these results can be replicated at other locations in future studies, it may be possible to develop an asthma/weather watch-warning system, based on the expected arrival of high-admissions air masses. PMID- 9412556 TI - Tolerance of volunteers to cyclosporine A-dilauroylphosphatidylcholine liposome aerosol. AB - Cyclosporine A (CsA) in liposomes of dilauroylphosphatidylcholine (DLPC), containing 118 micrograms of CsA/L of aerosol with a particle size of 1.6 to 1.7 micron diameter, was inhaled by 10 nonsmoking, normal volunteers each for 45 min. Aerosol was administered through an Aerotech II nebulizer (CIS-US, Inc., Bedford, MA) mouthpiece. Eight of the 10 volunteers had tracheal irritation and intermittent coughing following exposure. FEV1 and FVC values were mildly reduced, but returned to normal in 1 h. Blood chemical and hematologic values were unchanged at any time point after as opposed to before inhalation. Nine of the 10 volunteers later inhaled DLPC only, administered through the nebulizer mouthpiece. There was no change in FEV1 or FVC values, and there was no coughing or tracheal irritation. Subsequently, five of the volunteers who had previously had respiratory reactions inhaled CsA-DLPC liposome aerosol for 45-min, but through a mouth-only face mask. There was no tracheal irritation, coughing, or changes in spirometric measures. Blood concentrations of CsA at 15 min after the 45-min inhalation with a face mask averaged 83 +/- 42 ng/ml (mean +/- SD). At 24 h after treatment, CsA was undetectable in blood of the initial 10 volunteers. These studies indicate that CsA-DLPC liposome aerosol can be safely explored as a treatment for patients with moderately severe asthma. PMID- 9412557 TI - Enhanced thromboxane biosynthesis in patients with chronic obstructive pulmonary disease. The Chronic Obstructive Bronchitis and Haemostasis Study Group. AB - Thrombotic complications of pulmonary circulation occur in patients with chronic obstructive pulmonary disease (COPD). In the present study, we sought to evaluate in vivo platelet activation through the measurement of 11/dehydro-thromboxane (Tx) B2 TxA2 major metabolite in the urine, in 29 patients with COPD, compared with 29 sex- and age-matched healthy subjects. The urinary excretion of 11 dehydro-TxB2 was significantly higher in patients with COPD than in control subjects: median (range), 753 (277-4,409) and 275 (129-612) pg/mg creatinine, respectively; p < 0.0001). Moreover, 11-dehydro-TxB2 excretion was inversely related with arterial oxygen tension (rho = -0.46; p = 0.0145). In five of the 29 patients a short-term therapeutic course with oxygen supplementation induced a significant decrease of urinary 11-dehydro-TxB2 excretion: median range, 941 (452 2,640) to 445 (166-1,560) pg/mg creatinine. Moreover, selective inhibition of platelet cyclooxygenase activity by low-dose aspirin was associated with more than 90% inhibition of thromboxane metabolite excretion, demonstrating its being of platelet origin. Plasma levels of prothrombin fragment F1 + 2 were higher in patients than in control subjects (2.6 +/- 1.5 versus 0.9 +/- 0.4 nM, p = 0.0001). No relation between 11-dehydro-TxB2 excretion and plasma F1 + 2 levels was found. We conclude that platelet TxA2 biosynthesis is enhanced in patients with COPD and may be influenced by arterial oxygen tension changes. PMID- 9412558 TI - Administration of growth hormone to underweight patients with chronic obstructive pulmonary disease. A prospective, randomized, controlled study. AB - Patients with chronic obstructive pulmonary disease (COPD) often develop weight loss, which is associated with increased mortality. Recombinant human growth hormone (rhGH) treatment has been proposed to improve nitrogen balance and to increase muscle strength in these patients. The aim of this study was to assess the effects of rhGH administration on the nutritional status, resting metabolism, muscle strength, exercise tolerance, dyspnea, and subjective well-being of underweight patients with stable COPD. Sixteen patients attending a pulmonary rehabilitation program (age: 66 +/- 9 yr; weight: 77 +/- 7% of ideal body weight; FEV1: 39 +/- 13% of predicted) were randomly treated daily with either 0.15 IU/kg rhGH or placebo during 3 wk in a double-blind fashion. Measurements were made at the beginning (DO) and at the end (D21) of treatment and 2 mo later (D81). Body weight was similar in the two groups during the study, but lean body mass was significantly higher in the rhGH group at D21 (p < 0.01) and D81 (p < 0.05). The increase in lean body mass was 2.3 +/- 1.6 kg in the rhGH group and 1.1 +/- 0.9 kg in the control group at D21 and 1.9 +/- 1.6 kg in the rhGH group and 0.7 +/- 2.1 kg in the control group at D81. At D21, the resting energy expenditure was increased in the rhGH group (107.8% of DO, p < 0.001 compared with the control group). At D21 and D81, the changes in maximal respiratory pressures, handgrip strength, maximal exercise capacity, and subjective well-being were similar in the two groups. At D21, the 6-min walking distance decreased in the rhGH group ( 13 +/- 31%) and increased in the control group (+10 +/- 14%; p < 0.01). We conclude that the daily administration of 0.15 IU/kg rhGH during 3 wk increases lean body mass but does not improve muscle strength or exercise tolerance in underweight patients with COPD. PMID- 9412559 TI - Patterns of hospitalization in elderly patients with asthma and chronic obstructive pulmonary disease. AB - The purpose of this study was to describe the impact of asthma and chronic obstructive pulmonary disease (COPD) in the elderly on health care utilization. The Health Care Financing Administration (HCFA) file for the year 1984 through 1991 involving beneficiaries < or = 65 yr were searched for the diagnoses of asthma and COPD by ICD-9 codes. The study groups were created by determining the first admission for an exacerbation of either disease during each year from 1984 through 1991. Patients were identified by their social security number. The 1984 cohort consisted of 56,692 patients with asthma exacerbation and 162,899 with COPD exacerbation. The 1991 cohort consisted of 67,758 patients with asthma exacerbation and 131,974 patients with COPD exacerbation. In addition, the 1984 cohort was tracked by social security number for evidence of rehospitalization for either asthma or COPD through 1991. Length of hospitalization increased as patients grew older. The discharge rate to an independent living facility diminished as age increased. The use of convalescent and nursing homes or home health care after discharge more than doubled from 1984 through 1991. The utilization of health care resources by elderly patients with asthma and COPD is immense, both during hospitalization and after discharge. PMID- 9412560 TI - Sleep-disordered breathing and neuropsychological deficits. A population-based study. AB - The relationship of sleep-disordered breathing (SOB) to neuropsychological deficits was investigated with cross-sectional data from the Wisconsin Sleep Cohort Study, a population-based study of the natural history of SDB. A sample of 841 employed men and women ages 30 to 60 yr was studied by overnight polysomnography to assess the frequency of apneas and hypopneas per hour of sleep (apnea-hypopnea index, AHI). Prior to overnight polysomnography, the participants were given a battery of neuropsychological tests for functionally important capacities including motor skills, attention, concentration, information processing, and memory. Principal factor analysis of all the neuro-psychological test data revealed a psychomotor efficiency and a memory factor. Multiple regression analysis showed a significant negative association between logarithmically transformed AHI (LogAHI) and psychomotor efficiency score independent of age, gender, and educational status (p = 0.017). The relationship was not explained by self-reported sleepiness. No significant relationship was seen between LogAHI and memory score. In assessing the clinical significance of mild SDB, we estimate that an AHI of 15 is equivalent to the decrement in psychomotor efficiency associated with 5 additional yr of age, or to 50% of the decrement associated with hypnosedative use. PMID- 9412561 TI - Implementation of bronchoscopic techniques in the diagnosis of ventilator associated pneumonia to reduce antibiotic use. AB - In intensive care units, a large proportion of antibiotics are prescribed for presumed episodes of ventilator-associated pneumonia (VAP). VAP is usually diagnosed on a combination of clinical, radiographic, and microbiologic criteria with a high sensitivity but low specificity for VAP. As a result, patients may receive antibiotics unnecessarily. Specificity can be increased by the addition of quantitative cultures of samples of protected specimen brush (PSB) and bronchoalveolar lavage (BAL) to the diagnostic criteria. We prospectively analyzed the effects of implementation of PSB and BAL in the diagnosis of VAP on antibiotic prescription. PSB and/or BAL were performed in patients who fulfilled the clinical, radiographic, and microbiologic criteria for VAP. Based on quantitative cultures of PSB and/or BAL, patients were categorized into three groups: VAP microbiologically proven (Group 1; n = 72); clinical suspicion of VAP not confirmed microbiologically (Group 2; n = 66); and patients in whom bronchoscopy could not be performed (Group 3; n = 17). In Group 1, antibiotic therapy was instituted empirically in 40 patients (56%) (Group 1a) and after obtaining culture results in the other 32 patients (Group lb). Adjustment of therapy, based on culture results, occurred in 14 (35%) patients in Group la. In Group 2 empiric therapy was instituted in 34 (52%) patients (Group 2a) and dIscontinued within 48 h in 17 of them (50%). In Group 3, 17 (100%) patients were treated with antibiotics. Among the 66 patients in whom a clinical suspicion of VAP was not confirmed, only 18 (27%) were treated with antibiotics, and antibiotic therapy was withheld in 48 (35%) of 138 patients who underwent bronchoscopy. Withholding of antibiotic therapy had no negative effect on the recurrence of a clinical suspicion of VAP or on mortality rates. We conclude that addition of bronchoscopic techniques to the criteria for VAP may help to reduce antibiotic use. However, the definite benefits and cost-effectiveness of these techniques should be analyzed in a randomized study. PMID- 9412562 TI - Effects of inter-alpha-inhibitor in experimental endotoxic shock and disseminated intravascular coagulation. AB - We investigated the effects of human inter-alpha-inhibitor (I alpha I) on hemodynamics, oxygenation, and coagulation parameters in a porcine model of endotoxic shock. Four groups of six animals were studied: (1) control, (2) I alpha I group receiving 30 mg/kg I alpha I over 30 min, (3) LPS group receiving 5 micrograms.kg/min Escherichia coli endotoxin over 30 min, and (4) LPS + I alpha I group receiving 30 min after endotoxin 30 mg/kg/30 min I alpha I. We measured hemodynamic and oxygenation parameters, usual coagulation markers and plasma levels of thrombin-antithrombin complexes, antithrombin III activity, plasminogen activator tissue type, plasminogen activator inhibitor type 1, von Willebrand factor, tumor necrosis factor-alpha, and I alpha I at baseline and at 30, 60, 90, 120, 180, 240, and 300 min. In the I alpha I group, plasma I alpha I levels reached 447 +/- 23 mg/L just after injection and 287 +/- 39 mg/L at 300 min. I alpha I half-life was 7.3 +/- 1.9 h. In the IPS + I alpha I group, I alpha I plasma levels decreased more rapidly, reaching 260 mg/L at 300 min. Compared with the LPS group, administration of I alpha I normalized the mean arterial pressure and cardiac index, improved the LPS-induced pulmonary hypertension, and resulted in the blunted increase in blood lactate and oxygen extraction ratio. A significant decrease in thrombin-antithrombin complexes and plasminogen activator inhibitor type 1 levels were observed. There was no significant difference in plasma tumor necrosis factor-alpha levels. We concluded that in this hypodynamic model of endotoxin shock, I alpha I administration resulted in a marked improvement in the hemodynamic, oxygenation, and coagulation parameters. PMID- 9412563 TI - Alveolar and dead space volume measured by oscillations of inspired oxygen in awake adults. AB - Forced sinusoidal oscillations in the inspired concentration of a low-solubility inert gas can be used to measure airways dead space and alveolar volume. When inspired oxygen is oscillated about its mean value in the same way, the ratio between the amplitudes of the resulting end-expired and inspired oxygen oscillations is the same as that of an inert gas such as argon. Thus, oxygen forcing oscillations can be used to measure lung volume. In nine healthy spontaneously breathing adults, the FIO2 (mean FIO2 = 0.26, mean minute volume = 8.5 L/min) was forced to sinusoidally oscillate with an amplitude of +/- 0.04. The mean airways dead space measured using FIO2 oscillations with a forcing period of 3 min was 0.17 +/- 0.04 L, and the airways dead space measured by a single-breath C02 technique was no different at 0.19 +/- 0.03 L. An oxygen oscillation of the same period measured the mean end-expired alveolar volume at 3.1 +/- 0.7 L. Adding together the airways dead space and end-expired alveolar volume, obtained by the oxygen oscillation technique, provided a measure of FRC that at 3.3 +/- 0.7 L matched the FRC of 3.3 +/- 0.8 L measured by whole-body plethysmography. A third measure of FRC using a multiple-breath nitrogen washout technique gave a smaller volume of 3.00 +/- 0.85 L. The advantage of using FIO2 oscillations is that accurate FRC measurements can be made continuously, without interfering with the subject's natural breathing rhythm. PMID- 9412564 TI - Effects of noninvasive ventilation on pulmonary gas exchange and hemodynamics during acute hypercapnic exacerbations of chronic obstructive pulmonary disease. AB - Noninvasive positive pressure ventilation (NIPPV) can replace tracheal intubation in acute exacerbations of chronic obstructive pulmonary disease (COPD) with severe hypercapnic respiratory failure. However, the underlying mechanisms by which NIPPV improves pulmonary gas exchange are not known. We studied 10 male COPD patients (68 +/- 8 [SD] yr) with acute severe hypercapnic respiratory failure within 36 h after hospital admission. Measurements of pulmonary gas exchange, hemodynamics, and respiratory mechanics were done: (I) breathing spontaneously (baseline); (2) after 15 and 30 min of NIPPV with pressure support (inspiratory pressure = 12 +/- 2 cm H20, PEEP = 3 +/- 2 cm H20); and (3) 15 min after NIPPV withdrawal. Patients were ventilated using a full face mask, keeping FIO2 constant (0.23 +/- 0.02) in all conditions. Compared with baseline, during NIPPV (15 min) we observed a moderate increase in Pa02 (from 50 +/- 6 to 57 +/- 9 mm Hg; p < 0.05), and a fall in PaCO2 (from 66 +/- 10 to 59 +/- 10 mm Hg; p < 0.0001), but AaPO2 increased (from 39 +/- 13 to 48 +/- 13 mm Hg; p < 0.001). Breathing frequency decreased (from 26 +/- 5 to 19 +/- 3 breaths/min; p < 0.0001), tidal volume increased (from 311 +/- 42 to 520 +/- 133 ml; p < 0.0001), and minute ventilation increased (from 8.0 to 1.7 to 9.6 +/- 2.0 L/min; p < 0.05). Cardiac output fell during NIPPV in all patients (from 6.7 +/- 1.6 to 5.8 +/- 1.3 L/min; p < 0.0025) with no impact on mixed venous PO2. No substantial changes in VA/Q mismatching (multiple inert gas elimination technique) were observed. While oxygen uptake showed a trend to decrease, the respiratory exchange ratio (R) increased (from 0.78 +/- 0.17 to 0.90 +/- 0.22; p < 0.001). The effects of NIPPV were unchanged at 30 min compared with 15 min and were reversed after 15 min of NIPPV withdrawal. We conclude that improvement in respiratory blood gases during NIPPV is essentially due to higher alveolar ventilation (p < 0.001) and not to improvement in VA/Q relationships. The increase in AaPO2 was explained by the rise in R due to an increased clearance of body stores of C02 during NIPPV. Our results indicate that attainment of an efficient breathing pattern rather than high inspiratory pressures should be the primary goal to improve arterial blood gases during NlPPV in this type of patient. PMID- 9412565 TI - Endotoxin impairs agonist-induced calcium mobilization in rat mesangial cells. AB - We hypothesized that endotoxin would impair agonist-induced calcium (Ca2+) mobilization in rat mesangial cells, owing to the induction of nitric oxide synthase (NOS) and augmented nitric oxide (NO) synthesis. We measured basal and bradykinin-induced peak free cytosolic Ca2+ concentrations through microspectrofluorimetry with fura-2 in confluent mesangial cells, and assayed conditioned medium for nitrite accumulation. Prior to measurement, cells were incubated overnight in serum-supplemented medium, with or without endotoxin, 1 arginine, indomethacin, meclofenamate, or N omega-nitro-L-arginine methyl ester (L-NAME). Endotoxin (1 mg/ml) decreased bradykinin-induced peak Ca2+ responses by 35 to 60% (p < 0.0001) and increased nitrite accumulation > 6-fold (p < 0.01). Arginine supplementation further (> 9-fold, p < 0.0001) increased nitrite accumulation without changing the effect on Ca2+. Inhibition of NOS abolished increments in nitrite concentration but had no effect on impaired Ca2+ responses. Cyclooxygenase (COX) inhibitors, present during incubation with endotoxin, but not afterward, normalized bradykinin-stimulated calcium responses. Thrombin stimulated Ca2+ responses were similarly affected. We conclude that neither NO nor prostaglandins act directly to impair agonist-induced Ca2+ mobilization following endotoxin exposure; however, this effect may be an indirect effect of COX products, including reactive oxygen intermediates. PMID- 9412566 TI - Influence of ethnicity and gender on airway function in preterm infants. AB - While maximal expiratory flow at functional residual capacity, calculated from partial expiratory flow volume curves (V'maxFRC), is a valuable measure of peripheral airway function in infants, limited data are available in preterm infants despite their high prevalence of respiratory problems. To investigate the influence of gender and ethnic group, V'maxFRC and other indices of respiratory function were measured in 28 black and 28 white preterm infants (50% female in each group) at time of discharge from the neonatal unit (mean [SD] weight 2.36 [0.3] kg, postnatal age 19 [9] d). No infant had any history of cardiorespiratory disease and all were born to non-smoking mothers. V'maxFRC tended to be higher in girls than boys (115 versus 94 ml.s-1 [95% CI: -5; 47]) but there was no significant difference in this parameter between black and white infants (111 versus 98 ml.s-1 [95% CI of difference: -12; 40]). Respiratory resistance (Rrs) was significantly lower in black than white infants (95% CI: -2.9; -0.4 kPa.L 1.s) and tended to be lower in female than male infants (95% CI: -2.3; 0.2 kPa.L 1.s). Similarly, time to peak tidal expiratory flow as a proportion of total expiratory time (tPTEF:tE) was significantly longer in black than white (95% CI: 0.06, 0.20) and in female than male (95% CI: 0.02, 0.15) infants. These findings suggest that certain parameters of airway function may be influenced by both ethnic group and gender in preterm infants, both of which should therefore be taken mw account when investigating the effects of disease and/or therapeutic interventions in this group. PMID- 9412567 TI - Cold air challenge at age 6 and subsequent incidence of asthma. A longitudinal study. AB - The aim of this study was to assess the relation between bronchial hyperresponsiveness to dry, cold air at age 6 and the subsequent incidence of asthma. The cumulative incidence of newly diagnosed asthma between ages 6 and 11 among 360 children included in this study was 12.0%. Survival analysis showed that hyperresponsiveness to cold air at age 6 was associated with an increased risk of developing subsequent asthma (hazard ratio = 2.6, 95% CI = 1.2-5.4; p = 0.01). However, after adjusting for potential confounders, only mild wheezing at age 6 (adjusted hazard ratio 7.5, 95% CI = 3.6-15.9; p < 0.001) and skin test reactivity to allergens at age 6 (adjusted hazard ratio 3.6, 95% CI = 1.5-8.5; p < 0.01), but not hyperresponsiveness to cold air (adjusted hazard ratio = 0.9, 95% CI = 0.4-2.2; p = 0.8), remained significant predictors of subsequent development of asthma. These findings were substantially confirmed after stratifying for wheezing illnesses before age 3. We conclude that hyperresponsiveness to cold air at age 6 was associated with subsequent development of a diagnosis of asthma but this effect was not independent of atopy and mild wheezing at age 6. PMID- 9412568 TI - Airway function among Inuit primary school children in far northern Quebec. AB - The study of the prevalence and determinants of asthma and allergy in different populations may provide clues to their etiology. We describe airway function and its determinants among Inuit schoolchildren living in far Northern Quebec. We assessed the presence of airways hyperresponsiveness (AHR), defined as a 15% drop in FEV1 with exercise, airflow obstruction, as judged by a reduced FEV1/FVC, and atopy, as evidenced by skin test positivity to inhaled aeroallergens, among 509 Inuit aged mostly from 6 to 13 yr. Smoking by the children (31.9%) and their parents was common, including maternal smoking during pregnancy (79.5%). Atopy was found in only 5.3% of children. Apart from age, there were no significant associations between AHR and any of the determinants examined. Airflow obstruction was present among 7.7% of children and occurred most commonly among children with higher levels of salivary cotinine and in those with four or more lower respiratory illnesses in the first 2 yr of life. Asthma and atopy were uncommon in this population whereas evidence of chronic airflow obstruction was frequently found. Measures to reduce the spread of respiratory infection and prevention of smoking are likely to be of most benefit in improving respiratory health in these isolated communities. PMID- 9412569 TI - Influence of driving pressure on raised-volume forced expiration in infants. AB - The raised-volume forced-expiration technique measures infant lung function over an extended volume range. To improve comparisons between individuals and populations, we investigated the influence of jacket pressure on outcome variables in 21 infants. To quantify pressure transmitted from the jacket to the pleural space at a given lung volume, the jacket was inflated against an occluded airway, and the increase in pressure at the mouth was measured. Flow-volume curves were recorded at transmitted pressure (Ptrans) values ranging from 0 to 41.9 cm H20. The effect of Ptrans on the FEV measures of FEV0.5, FEV0.75, and FVC, and on the forced expiratory flow measures of FEF25%, FEF50% and FEF75% was assessed. At Ptrans values between 0 to 20 cm H20, a significant positive relationship existed between transmitted pressure (Ptrans) and all outcome variables except FVC. At higher Ptrans values, all outcome variables demonstrated pressure independence, with the exception of FEF25% (which remained positive) and FVC (which was negative in a subgroup of wheezy infants). FEF75% values tended to decrease at Ptrans values > 25 cm H20. At Ptrans values between 20 and 25 cm H20, most outcome variables are pressure independent. This range is therefore the most suitable for use with the raised-volume forced expiration technique. PMID- 9412570 TI - Beryllium-stimulated release of tumor necrosis factor-alpha, interleukin-6, and their soluble receptors in chronic beryllium disease. AB - Chronic beryllium disease (CBD) provides a model system in which to evaluate the antigen-stimulated, cell-mediated, immune response that leads to granulomatous lung disease. We hypothesized that beryllium salts would stimulate bronchoalveolar lavage (BAL) cell release of tumor necrosis factor-alpha (TNF alpha) and lnterleukin-6 (IL-6), and their soluble receptors, soluble TNF receptor I (sTNF RI), sTNF RII, and sIL-6R and that chronic exposure to antigen would increase production of soluble receptors in the serum and BAL fluid (BALF) of beryllium-sensitized and CBD patients. We have demonstrated (1) similar constitutive TNF-alpha, IL-6, and soluble receptor production by control subjects and CBD patients, (2) a BeSO4-stimulated increase in TNF-alpha and IL-6 production by CBD-derived BAL cells, and (3) a BeSO4-induced decrease in sTNF RII production by BAL cells from control subjects. We measured increased serum sTNF RI and serum and BALF sIL-6R in beryllium-sensitized subjects and increased sTNF RI and RII in serum and sIL-6R and sTNF RII and BALF in CBD patients. These changes correlated with pulmonary lymphocytosis and clinical measures of disease severity, indicating that soluble receptors may reflect disease status. PMID- 9412571 TI - Exhaled nitric oxide and bronchoalveolar lavage nitrite/nitrate in active pulmonary sarcoidosis. AB - Increased exhaled nitric oxide (NO) may reflect respiratory tract inflammation in untreated asthmatics. We compared exhaled NO and bronchoalveolar lavage (BAL) nitrate/nitrite (NO3-/NO2-) in 10 patients who had untreated, active pulmonary sarcoidosis with those of normal control subjects. Exhaled NO concentrations, determined by chemiluminescence, were similar in patients and control subjects (peak NO concentration of patients [mean +/- SD]: 13.6 +/- 5.9 parts per billion [ppb], peak NO concentration of control subjects: 11.2 +/- 5.7 ppb, p = 0.32; mean alveolar NO concentration of patients: 7.8 +/- 4.4 ppb, mean alveolar NO concentration of control subjects: 7.1 +/- 4.2 ppb, p = 0.70; end-tidal NO concentration of patients: 6.9 +/- 4.5 ppb, end-tidal NO concentration of control subjects: 6.6 +/- 4.0 ppb, p = 0.60). BAL NO2- was assayed using a modified Griess reaction after reduction of NO3- to NO2-. There was no significant difference in mean BAL NO2- concentrations, expressed as nanomoles per milliliter of epithelial lining fluid (patients: 544 nmol/ml, control subjects: 579 nmol/ml, p = 0.81) or as nanomoles per milliliter of BAL fluid (patients: 6.7 nmol/ml, control subjects: 5.7 nmol/ml, p = 0.41). These data suggest that excess NO generation does not accompany the respiratory tract inflammation of pulmonary sarcoidosis. PMID- 9412572 TI - Antioxidative and clinical effects of high-dose N-acetylcysteine in fibrosing alveolitis. Adjunctive therapy to maintenance immunosuppression. AB - In fibrosing alveolitis (FA), activated phagocytes cause excessive oxidative stress in the lower respiratory tract. Additionally, levels of glutathione, a major antioxidant of the human lung, are markedly reduced. Since N-acetylcysteine (NAC) is a known precursor for glutathione synthesis, we investigated the effect of NAC on redox balance and lung function in FA. Eighteen patients with an established diagnosis of FA were treated with 600 mg NAC three times daily for 12 wk in addition to their latest immunosuppressive therapy. Before and after NAC therapy, pulmonary function tests (PFTs) and bronchoalveolar lavage (BAL) were performed. BAL fluid was analyzed with regard to cell differential, glutathione status, and methionine sulfoxide content of BAL proteins (Met(O)), as an indicator of oxidative stress at the alveolar surface. There was an increase of total glutathione (GSHt = GSH +/- 2 x GSSG: 3.43 +/- 0.30 microM versus 4.20 +/- 0.66 microM, p < 0.05) and of reduced glutathione (GSH: 2.58 +/- 0.24 microM versus 3.42 +/- 0.54 microM, p < 0.005) in native BAL fluid and in the epithelial lining fluid (GSHt: 267.3 +/- 26.0 microM versus 367.1 +/- 36.0 microM, p < 0.005; GSH: 204.5 +/- 20.7 microM versus 302.9 +/- 32.2 microM, p < 0.005). The increase of GSH was accompanied by a decrease of Met(O) (6.83 +/- 0.71% versus 4.60 +/- 0.40%, p < 0.005). PFTs significantly improved during NAC treatment. We conclude that high-dose NAC significantly improved the antioxidant screen of the lungs by elevating GSH levels. Moreover, the decrease of Met(O) levels indicated an antioxidant effect at the alveolar surface. These biochemical changes were accompanied by an improvement of PFTs in patients under maintenance immunosuppression. NAC supplementation should, therefore, be considered as an adjunct therapy for FA. PMID- 9412573 TI - Plasma 3-nitrotyrosine in cigarette smokers. AB - Peroxynitrite has been associated with increased oxidative reactions and DNA damage in inflamed tissues as it may cause a reduction of plasma antioxidants as well. Nitration of tyrosine residues of proteins leads to the production of 3 nitrotyrosine (NTYR), which may be considered as a marker of NO.-dependent oxidative damage. We developed a highly sensitive method to detect NTYR in human plasma and tested it in cigarette smokers and in healthy control subjects. Peripheral venous blood (10 ml) was obtained in 20 healthy, asymptomatic cigarette smokers (13 males, 7 females; age: 49 +/- 11 yr) and in 18 healthy nonsmokers (10 males and 8 females; age: 36 +/- 6 yr). In smokers, plasma nicotine, cotinine, and expired CO levels were measured. NTYR was determined with a sequential HPLC/gas chromatography-thermal energy analysis (GC-TEA) technique. The total plasma Trolox-equivalent antioxidant capacity (TEAC) was also measured using metmyoglobin as peroxidase and a phenothiazine as a radical donor. NTYR was detectable (detection limit: 0.02 ng/injection) in 11 smokers (mean +/- SD: 1.60 +/- 1.24 ng/mg protein) and in two nonsmokers (1.10 and 1.20 ng/mg protein, respectively). NTYR was not associated with nicotine and cotinine levels or expired CO in smokers. Plasma TEAC in smokers was significantly lower (0.43 +/- 0.38 mM) than in nonsmokers (1.42 +/- 0.3 mM; p < 0.001) and showed a biphasic, negative relationship with NTYR (r = 0.96, p < 0.001). This highly sensitive HPLC/GC-TEA method for detection and quantitation of plasma NTYR may be used for monitoring oxidative reactions associated with tobacco smoking. This assay might be incorporated into molecular epidemiologic studies for lung chronic inflammatory and neoplastic disorders in which exposure to oxidants may be an important risk factor. PMID- 9412574 TI - Community-acquired pneumonia in the elderly. Clinical and nutritional aspects. AB - Community-acquired pneumonia (CAP) in the elderly has a different clinical presentation than CAP in other age groups. Confusion, alteration of functional physical capacity, and decompensation of underlying illnesses may appear as unique manifestations. Malnutrition is also an associated feature of CAP in this population. We undertook a study to assess the clinical and nutritional aspects of CAP requiring hospitalization in elderly patients (over 65 yr of age). One hundred and one patients with pneumonia, consecutively admitted to a 1,000-bed teaching hospital over an 8-mo period, were studied (age: 78 +/- 8 yr, mean +/- SD). Nutritional aspects and the mental status of patients with pneumonia were compared with those of a control population (n = 101) matched for gender, age, and date of hospitalization. The main symptoms were dyspnea (n = 71), cough (n = 67), and fever (n = 64). The association of these symptoms with CAP was observed in only 32 patients. The most common associated conditions were cardiac disease (n = 38) and chronic obstructive pulmonary disease (COPD) (n = 30). Seventy-seven (76%) episodes of pneumonia were clinically classified as typical and 24 as atypical. There was no association between the type of isolated microorganism and the clinical presentation of CAP, except for pleuritic chest pain, which was more common in pneumonia episodes caused by classical microorganisms (p = 0.02). This was confirmed by a multivariate analysis (relative risk [RR] = 11; 95% confidence interval [CI]: 1.7 to 65; p = 0.0099). The prevalence of chronic dementia was similar in the pneumonia cohort (n = 25) and control group (n = 18) (p = 0.22). However, delirium or acute confusion were significantly more frequent in the pneumonia cohort than in controls (45 versus 29 episodes; p = 0.019). Only 16 patients with pneumonia were considered to be well nourished, as compared with 47 control patients (p = 0.001). Kwashiorkor-like malnutrition was the predominant type of malnutrition (n = 65; 70%) in the pneumonia patients as compared with the control patients (n = 31; 31%) (p = 0.001). The observed mortality was 26% (n = 26). Pleuritic chest pain is the only clinical symptom that can guide an empiric therapeutic strategy in CAP (typical versus atypical pneumonia). Both delirium and malnutrition were very common clinical manifestations of CAP in our study population. PMID- 9412575 TI - Impact of immigration on tuberculosis infection among Canadian-born schoolchildren and young adults in Montreal. AB - We conducted a cross-sectional tuberculin survey among non-BCG-vaccinated Canadian-born schoolchildren in grades 6 and 10, health professional students, and young adult workers, to estimate the association of tuberculin reactions with indices of contact with tuberculosis. Participants underwent simultaneous tuberculin testing with PPD-T (standard) and PPD-B (from M. intracellulare). Exposure was estimated from questionnaire responses, group, aggregate census, and tuberculosis incidence data. Of 3,710 participants, 88 (2.4%) had positive tuberculin reactions, i.e., of 10+ mm. Positive tuberculin reactions were rarely associated with larger reactions to PPD-B, but were associated with older age (adjusted odds ratio for each 5 years: 1.5 [95% confidence interval, 1.3, 1.8]), household contact (4.2 [1.4, 12.7]), and population group (health professional versus all others: 0.6 [0.3, 1.0]). Estimated annual risk of infection declined by 3% per year. Tuberculin reactions were not associated with any indices of contact in school, work or neighborhood settings with foreign-born from tuberculosis endemic areas, nor with tuberculosis in Canadian-born. There was no evidence of transmission of tuberculosis from affected high risk sub-groups in Montreal to the general population working or attending school. PMID- 9412576 TI - Community-acquired aspiration pneumonia in intensive care units. Epidemiological and prognosis data. AB - Over a 9-yr period, among 505 patients exhibiting severe community-acquired pneumonia and admitted into a total of six medical ICUs in the north of France, we collected 116 patients (23%) meeting the usual criteria for aspiration pneumonia. Main medical grounds of ICU admission were respiratory distress in 54 patients and neurological disturbances in 62 patients. The main underlying risk factor for aspiration pneumonia was drug overdose (39%). Mechanical ventilation was required for 73 patients. Initial shock was present in 15 patients. Pulmonary involvement was bilateral in 27 patients. There were 94 aerobic organisms isolated from 70 patients (60%), the most frequent being gram-negative bacilli (n = 38), Staphyloccus spp. (n = 27) and Streptococcus pneumoniae (n = 22). Overall mortality was 22%, but only 11 (11%) deaths were directly or indirectly related to aspiration pneumonia. Stepwise multivariate analysis identified four independent predictors of mortality: ineffective initial antimicrobial therapy (p = 0.0001), positive initial blood culture (p = 0.0001), hospital-acquired lower respiratory tract superinfections (p = 0.0054), and use of inotropic support (p = 0.0078). The importance of prevention of hospital-acquired superinfections and permanent optimization of our antimicrobial strategies warranting efficacy of the initial antimicrobial therapy is underlined. PMID- 9412577 TI - Association of expression of metalloproteinases and their inhibitors with the metastatic potential of squamous-cell lung carcinomas. A molecular and immunohistochemical study. AB - Matrix metalloproteinases (MPs) constitute a family of proteolytic enzymes (proteases) that degrade extracellular matrix (ECM) and promote the local or metastatic potential of carcinoma cells, and whose action is restrained by special inhibitors (metalloproteinase inhibitors; MIs). We assessed the role of the MPs stromelysin-3 (STR-3), putative metalloproteinase-1 (PUMP-I), and the gelatinases of molecular weights 72 kDa and 92 kDa, as well as the role of their inhibitors tissue inhibitor of metalloproteinase-1 (TIMP-1) and TIMP-2, as markers of metastatic potential in 25 fresh biopsies of squamous-cell lung carcinomas (SCLCs). We examined levels of messenger ribonucleic acid (mRNA) expression for these MPs and inhibitors through Northern blot analysis in 10 carcinomas of high-to-moderate differentiation without lymph-node involvement, and in 15 infiltrative carcinomas of moderate-to-low differentiation with lymph node involvement. Five cases with significant epithelial atypia and five samples with normal mucosa were used as controls. Expression of STR-3 and TIMP-2 was also assessed immunohistochemically with the avidin-biotin-complex (ABC) technique. We noticed a progressive increase in the expression levels of MPs, especially of STR 3, and of TIMP-2, from the stage of epithelial atypia to the detection of carcinoma, finding the highest values of these substances among carcinomas of low differentiation with nodal metastases. These findings were also confirmed with immunohistochemical analysis. Our results suggest that there is a significant association of the expression of MPs and MIs with both the local and metastatic potential and the degree of cellular differentiation of SCLC, and that this association is clinically important because of its prognostic and therapeutic implications. PMID- 9412578 TI - Simultaneous or delayed administration of hepatocyte growth factor equally represses the fibrotic changes in murine lung injury induced by bleomycin. A morphologic study. AB - Hepatocyte growth factor (HGF) is a humoral mediator of epithelial-mesenchymal interactions, acting on a variety of epithelial cells as mitogen, motogen, and morphogen. Exogenous HGF acts as a hepatotrophic factor and a renotrophic factor during experimental injury. To investigate whether HGF has a pulmotrophic function, human recombinant HGF was administered to C57BL/6 mice with severe lung injury by bleomycin (BLM). Low dose simultaneous and continuous administration of HGF (50 micrograms/mouse/7 d) with BLM (100 mg/mouse/7 d) repressed fibrotic morphological changes at 2 and 4 wk. Ashcroft score showed a significant difference in lung fibrosis with and without HGF at 4 wk (3.7 +/- 0.4 versus 4.9 +/- 0.3, p < 0.05). Furthermore, either simultaneous or delayed administration of high dose HGF (280 micrograms/mouse/14 d) equally repressed fibrotic changes by BLM when examined at 4 wk (Ashcroft score: 2.6 +/- 0.4 and 2.4 +/- 0.2 versus 4.1 +/- 0.2, p < 0.01). Hydroxyproline content in the lungs was significantly lower in mice with either simultaneous or delayed administration of high dose HGF as compared to those administered BLM alone (121.8 +/- 8.1% and 113.2 +/- 6.2% versus 162.7 +/- 4.6%, p < 0.001). These findings indicate that exogenous HGF acts as a pulmotrophic factor in vivo and prevents the progression of BLM-induced lung injury when administered in either a simultaneous or delayed fashion. HGF may be a potent candidate to prevent or treat lung fibrosis. PMID- 9412579 TI - CD23 deficient mice develop allergic airway hyperresponsiveness following sensitization with ovalbumin. AB - The low affinity receptor for IgE (CD23) is reported to regulate immune and inflammatory events and as a result, it may have a role in the development of allergic airway inflammation and hyperresponsiveness (AHR). To test this hypothesis CD23-deficient mice were studied following different modes of allergic sensitization. Mice were actively sensitized either intraperitoneally with ovalbumin (OA)/alum or via the airways (10 days exposure to OA aerosol with no adjuvant). Passive sensitization was performed by intravenous injections of OA specific IgE. Airway responsiveness, serum IgE and IgG levels were assessed together with airway inflammation. Passive sensitization followed by airway challenges resulted in increased OA-specific lgG and IgE in the serum of wild type mice only, while both the CD23+/+ and CD23-/- groups developed tracheal smooth muscle hyperresponsiveness to electrical field stimulation, indicating that IgE/CD23-mediated immune functions may not be necessary for the development of allergic changes. Active sensitization of both CD23-/- and CD23+/+ mice resulted in increased serum levels of OA-specific IgE and lgG, airway eosinophilia and significant AHR when compared with nonsensitized mice. The genetic deficiency of CD23-/- mice not only failed to prevent but was associated with a significant increase of these responses. These results indicate that CD23 may not be essential for the development of allergen-induced AHR and further, that its presence may have some inhibitory effects on the allergic response. PMID- 9412580 TI - Allergens of Aspergillus fumigatus and Candida boidinii share IgE-binding epitopes. AB - From an Aspergillus fumigatus complementary deoxyribonucleic acid (cDNA) library displayed on phage surface, an allergen formally termed rAsp f 3 was cloned. The open-reading frame of the cloned gene for the allergen encodes a protein of 168 amino acids with a predicted molecular mass of 18.5 kD, showing 36% identity and 58% similarity to two peroxisomal membrane proteins of Candida boidinii. Recombinant Asp f 3 was expressed as a [His]6-tagged fusion protein in Escherichia coil at yields of 30 mg/L, and was purified by Ni(2+)-chelate chromatography. In an enzyme-linked immunosorbent assay (ELISA), serum IgE antibody reactivity to rAsp f 3 could be detected in 72% of 89 individuals sensitized to A. fumigatus, demonstrating that the protein represents a major allergen of the mold. IgE specific to rAsp f 3 and the two recombinant Candida proteins was further demonstrated by IgE-immunoblot analysis. IgE binding to rAsp f 3 could be inhibited in the ELISA by adding either of the recombinant Candida peroxisomal proteins to sera containing IgE directed against Asp f 3. Taken together, these observations prove that the Asperigillus allergen and the two Candida proteins share IgE-binding epitopes. PMID- 9412581 TI - Increased VIP-positive nerve fibers in the mucous glands of subjects with chronic bronchitis. AB - The presence and distribution of neuropeptide-containing nerves within bronchial surgical specimens has been investigated in bronchitic (n = 12) and in nonbronchitic subjects (n = 7). Lung tissue, obtained from patients undergoing thoracotomy for limited lung lesions, was processed immediately and analyzed for nerves using the streptavidin-biotin complex peroxidase method with antisera to the neural marker protein gene product 9.5 (PGP 9.5) and the neuropeptides vasoactive intestinal peptide (VIP), substance P (SP), calcitonin-gene related peptide (CGRP). There were no significant differences between the two groups with respect to the density of PGP 9.5-, SP-, or CGRP-positive nerves in both the locations assessed (smooth muscle layer and glands). The density of VIP-positive nerves was significantly higher in the glands of bronchitic than in nonbronchitic subjects. A negative relationship was found between the presence of airway inflammation, as indexed by mononuclear cell tissue infiltration, and the density of PGP 9.5-positive nerves in both smooth muscle and glands. Likewise, a relationship was found between the smoking history (packs/yr and age of onset of smoking) and the density of VIP-positive nerves in glands. These findings support a role for VIP in the hallmark of chronic bronchitis, i.e., sputum production. PMID- 9412582 TI - Neutrophil apoptosis in the acute respiratory distress syndrome. AB - Little is known about neutrophil (PMN) apoptosis in the acute respiratory distress syndrome (ARDS). We uses morphologic criteria to count apoptotic PMN in bronchoalveolar lavage fluid (BAL) of 35 patients on Days 1, 3, 7, 14, and 21 of ARDS and 13 patients on Days 1 and 3 of risk for ARDS. We found that the proportion of apoptotic PMN in BAL was low throughout the course of ARDS. There was no significant difference between the percentage of apoptotic PMN in patients at risk and patients with established ARDS or between patients who lived (2.4%) and patients who died (1.8%). When normal human PMN were incubated in ARDS BAL, a significantly lower proportion became apoptotic (50 +/- 4%), as compared with PMN incubated in lavage fluid from normal volunteers (76 +/- 7%, p < 0.05). This antiapoptotic effect of ARDS BAL was blocked by immunodepleting BAL of G-CSF and GM-CSF. We conclude that the proportion of apoptotic PMN recovered from the lungs of patients with ARDS is low throughout the course of ARD S. Furthermore, BAL from patients with ARDS prolongs survival of normal human PMN in vitro, and this effect is partially mediated by G-CSF and GM-CSF. PMID- 9412583 TI - Increased gelatinolytic activity in bronchoalveolar lavage fluid in stable lung transplant recipients. AB - Proteolytic enzymes have been proposed to play a role in the pathogenesis of various inflammatory pulmonary diseases accompanied by parenchymal remodeling. To assess the role of inflammatory cells and proteolytic enzymes in the development of chronic allograft rejection after lung transplantation, bronchoalveolar lavage fluid (BALF) samples from clinically stable lung transplant (LT) recipients (i.e., without evidence of active infection or rejection), heart transplant (HT) recipients, and healthy volunteers (NL) were analyzed for total white blood cell (WBC) count and differential cell count, along with gelatinolytic/type IV collagenolytic activity. The LT group displayed a significantly increased total WBC count, neutrophil count, and percent neutrophils compared with the NL group, confirming the presence of inflammation. Furthermore, gelatin zymography revealed a significant increase in activity of the 72 and 92 kD gelatinases in the LT group compared with the NL group. A positive correlation existed between neutrophil counts and the increase in proteolytic activity. Immunosuppressive therapy did not account for the findings, since no significant difference in cell counts or proteolytic activity existed between the NL and HT control groups. These findings, together with those of others that relate chronic lung allograft dysfunction to an increase in BALF neutrophils and collagen matrix remodeling, collectively indicate that up-regulated proteolytic activity may have a role in chronic rejection after lung transplantation. PMID- 9412584 TI - MMP and TIMP expression pattern in pleural effusions of different origins. AB - The matrix metalloproteinases (MMP) are proteolytic enzymes that are essentially involved in the turnover of the extracellular matrix (ECM). Their activity is counterbalanced by specific antagonists, the tissue inhibitors of metalloproteinases (TIMP). In this study, we sought to analyze the expression of MMP and TIMP isoforms in pleural effusions from 88 patients. We compared MMP and TIMP isoform expression in transudates (n = 21) and exudates (n = 67), the latter divided into exudates of paraneoplastic (n = 46) or parainfectious (n = 21) origin. Zymographic and Western blot analyses revealed constant expression of interstitial collagenase (MMP-1), gelatinase-A (MMP-2), and TIMP-1 in all 88 samples. In contrast, analyses of gelatinase-B (MMP-9) demonstrated a specific expression pattern, with high expression in exudates and lack of expression in transudates. Neutrophil collagenase (MMP-8) was detected in trace amounts, and correlated with the number of neutrophils in the effusion. Low levels of TIMP-2 were detected only in exudates and not in transudates. Quantitative analysis of the expression ratio of gelatinase-B to gelatinase-A revealed statistically significant differences between effusions of different origin. The ratio was highest in exudates of paraneoplastic origin and lowest in transudates. Our data thus suggest that interstitial collagenase, gelatinase-A, and TIMP-1 play a role in homeostasis of the pleural space in vivo as constitutively expressed proteins, whereas gelatinase-B and TIMP-2 expression are induced in specific disease states. These observations contribute to the understanding of the pathophysiology of pleural effusions, and may help to characterize and possibly distinguish effusions of different origin. PMID- 9412585 TI - Development of a suicide gene as a novel approach to killing Mycobacterium tuberculosis. AB - The increase in multidrug-resistant tuberculosis and high mortality among those co-infected with HIV-1 necessitates new therapeutic approaches directed at Mycobacterium tuberculosis. We hypothesized that a dominant-negative mutation in the DNA-dependent RNA polymerase gene would inhibit transcription of all genes by blocking access of the wild-type enzyme to promoters. An evolutionarily invariant lysine was substituted with arginine by site-directed mutagenesis in the rpoB gene. The dominant-negative rpoB gene product inhibited a transposon-derived kanamycin-resistance gene in both M. smegmatis and M. tuberculosis H37Rv, leading to growth inhibition of the mycobacteria on solid media containing kanamycin. The dominant-negative mutant rpoB gene is a potential suicide gene especially for the treatment of multidrug-resistant tuberculosis once a delivery strategy is also developed. PMID- 9412586 TI - GM-CSF gene expression is normal but protein release is absent in a patient with pulmonary alveolar proteinosis. AB - Pulmonary alveolar proteinosis (PAP) is a rare disease characterized by an excessive accumulation of surfactant lipids and proteins in the alveolar space. In mice with a homozygous deletion of granulocyte macrophage-colony stimulating factor (GM-CSF), their phenotype mimics PAP. To evaluate whether the knockout mouse model mimics human disease, we evaluated GM-CSF expression in alveolar macrophages from a patient with PAP. We performed multiple whole lung lavages on a patient with PAP, and cultured BAL cells in the presence or absence of LPS. In contrast to the GM-CSF knockout mouse, human BAL cells from a patient with PAP expressed mRNA for GM-CSF following LPS stimulation. However, similar to the knockout mouse, GM-CSF protein release from BAL cells was undetectable with or without LPS. BAL cells from normal human controls released GM-CSF in abundance after LPS stimulation. In BAL cells from the patient with PAP, neutralization of interleukin-10 (IL-10) by anti-IL-10 antibody, resulted in enhanced GM-CSF production. Thus, alveolar macrophages from a PAP lung have deficient GM-CSF production analogous to the GM-CSF knockout mice; in contrast, human cells from a PAP lung have an intact GM-CSF gene. This case report illustrates an important difference between the knockout mouse model of PAP and the human disease. PMID- 9412587 TI - Recurrence of desquamative interstitial pneumonia after lung transplantation. PMID- 9412588 TI - Inhalation of the nitric oxide synthase cofactor tetrahydrobiopterin in healthy volunteers. AB - Pulmonary endothelial dysfunction is the hallmark of acute lung injury. Impaired pulmonary endothelial nitric oxide (NO) production in this event has been described. Tetrahydrobiopterin (BH4) is an essential cofactor for NO synthase and modulator of its activity. At high local concentrations, BH4 provokes local vasodilation in vivo in healthy individuals. At lower concentrations, BH4 selectively and locally restores disturbed NO-dependent vasodilation in patients with endothelial dysfunction. In this preliminary study, we therefore investigated the feasibility of BH4 inhalation in five healthy human volunteers. Inhalation of buffered, aqueous BH4-dihydrochloride solution was well tolerated; despite the buffer, BH4 stability was completely preserved. Resorption of inhaled BH4 was demonstrated by significantly increased BH4 levels in plasma and urine. Inhaled BH4 did not alter pulmonary function and had no effect on systemic hemodynamic values. Our data demonstrate that inhalation is a novel method for local BH4 administration, offering a basic therapeutic tool for investigation of restoration of impaired NO-dependent vasodilation due to pulmonary endothelial dysfunction. PMID- 9412589 TI - Electrocardiographic prediction of hyperinflation in children. AB - The mean frontal P wave axis in an electrocardiogram (ECG), which reflects the atrial orientation in the thorax, is altered by the relationship between atria and the diaphragm and, therefore, by hyperinflation. To examine this relationship, 102 children (ages 6-18) with asthma were prospectively studied. Lung volumes were estimated by plethysmography and a standard ECG obtained before and after bronchodilator. The mean thoracic gas volume (TGV) was 120.7 +/- 2.1% of predicted and the mean P axis was 54.9 +/- 1.5 degrees. Sixty-two subjects (61%) had a "vertical" P axis (> or = 60 degrees). Of 27 subjects with moderate or severe hyperinflation (TGV > or = 130% predicted), 23 (85%) had a vertical P axis. As a measure of significant hyperinflation, a vertical P axis had a sensitivity of 85%, specificity of 49%, positive predictive value of 38% and a negative predictive value of 90%. After nebulized albuterol, the mean TGV decreased to 96.4 +/- 1.3% predicted and the mean P axis decreased by 7.1 +/-1.6 degrees. Sixty-two of 76 subjects (82%) with > or = 15% decrease in TGV also had a decrease in P axis, and 62/67 subjects (93%) with a decrease in P axis also had > or = 15% decrease in TGV. The sensitivity was 82%, specificity 81%, and positive predictive value 93% for a decrease in P axis as a measure of decrease in TGV. A vertical P axis combined with a decrease in P axis after bronchodilator is highly sensitive and predictive for hyperinflation in children. PMID- 9412590 TI - Attributes of ATS documents that guide clinical practice. Recommendations of the ATS Clinical Practice Committee. PMID- 9412591 TI - Lyme disease and facial nerve palsy. PMID- 9412592 TI - Is phototherapy in neonates a risk factor for malignant melanoma development? AB - OBJECTIVE: To determine whether phototherapy of the newborn's sensitive skin could be a risk factor for malignant melanoma. DESIGN: Retrospective study from 1973 to 1992. SETTING: The Swedish Medical Birth Registry and the Swedish Cancer Registry. PATIENTS: Thirty cases of childhood malignant melanoma and 120 matched controls. INTERVENTION: None. MAIN OUTCOME MEASURES: Data in birth and cancer reports and from death certificates in Sweden. RESULTS: None of the patients with childhood malignant melanoma had received phototherapy, in comparison with 11 of the controls. This difference was not significant (P = .08; Fisher exact test). CONCLUSION: This preliminary report shows no significant risk of developing childhood malignant melanoma after phototherapy of the skin in neonates with hyperbilirubinemia. However, the median follow-up time was only 18 years. PMID- 9412593 TI - Cigarette promotional items in public schools. AB - OBJECTIVES: To assess the prevalence of ownership of cigarette promotional items (CPIs) by rural northern New England students and to examine the association between CPI ownership and smoking behavior. DESIGN AND SETTING: Voluntary, self administered survey of 1265 sixth- through 12th-grade students representing 79% to 95% of all students attending 5 rural New Hampshire and Vermont public schools in October 1996. We examined the association between ownership of a CPI and smoking behavior through regression models and conducted a sensitivity analysis on the findings. MAIN OUTCOME MEASURES: Adjusted odds of being a smoker (lifetime use of > or = 100 cigarettes) and, among never smokers and experimental smokers, adjusted cumulative odds of having higher levels of smoking uptake given CPI ownership. RESULTS: One third of students owned a CPI. Prevalence of ownership did not vary by grade or sex, but was higher among poor-to-average school performers (45.0% vs 21.0% for excellent school performers, P < .001) and children whose friends and family members smoked (43.4% vs 13.8% for students with no family members or friends smoking, P < .001). Cigarette promotional items included articles of clothing (T-shirts, hats, backpacks, and jackets), smoking paraphernalia (lighters and ashtrays), camping gear, and electronics. More than half of CPIs (58.2%) bore the Marlboro logo, and almost one third (31.7%) bore the Camel logo. These items were obtained directly from catalogs or vendors 22.4% of the time. Whereas only 4.5% of students reported bringing a CPI to school with them the day of the survey, 44.5% reported seeing such an item at school the day of the survey. After controlling for confounding factors, such as having friends who smoke, students who owned CPIs were 4.1 times more likely to be smokers than those who did not own CPIs (95% confidence interval [CI], 3.1-5.5). Never and experimental smokers (n = 1008) who owned CPIs were more likely to be in a higher category on the smoking uptake index in grades 6 (cumulative odds ratio [OR = 5.7, 95% CI, 1.9-16.8), 7 (OR = 1.8, 95% CI, 0.9-3.7), 8 (OR = 2.3, 95% CI, 1.1 4.8), and 9 (OR = 2.1, 95% CI, 1.1-3.9), periods when children are most vulnerable to initiating cigarette use. A sensitivity analysis indicated that an unmeasured confounder of CPI ownership and smoking was unlikely to alter our conclusions. CONCLUSIONS: Cigarette promotional items are owned by one third of students in these rural northern New England schools. These items are highly visible in the public school setting, and their ownership is strongly associated with initiation and maintenance of smoking behavior. These data lend support to a ban on CPIs to be included in US Food and Drug Administration regulations to prevent tobacco use among US youth. PMID- 9412594 TI - Trends in anabolic-androgenic steroid use among adolescents. AB - OBJECTIVE: To examine the trends in anabolic steroid use among adolescents in the United States between 1988 and 1996. DESIGN: Computerized and manual literature searches were performed, and the resultant local, state, and national cross sectional surveys of illicit anabolic steroid use by adolescents were reviewed. Trends in steroid use were evaluated using state and national studies administered in multiple periods. Various sampling procedures were employed, and all surveys used anonymous questionnaires. The national studies used for this analysis included the Monitoring the Future (MTF) study, the national component of the Youth Risk and Behavior Surveillance System, and the National Household Survey on Drug Abuse. SETTING: Most of the surveys described were self administered in school classrooms. The National Household Survey on Drug Abuse was administered in the respondent's home. PARTICIPANTS: Most survey respondents were junior high an high school male and female students aged 12 to 18 years. RESULTS: Individual state studies (ie, a single point in time) provide evidence of continued steroid use throughout the United States despite educational and legal interventions. The findings of multiyear state-level studies show a decrease in lifetime steroid use rates between 1988 and 1994 for male and female adolescents, although no tests of statistical significance were conducted. At the national level, a significant decline (P < .01) in lifetime steroid use has taken place from 1989 to 1996 for male and female students (MTF data). However, since 1991, lifetime steroid use by male students, as measured by 2 of the 3 national surveys, has been generally stable. The third survey, MTF, shows a significant decrease (P < .05) in use from 1991 to 1996. Likewise, from 1991-1996 use of anabolic steroids during the past year (MTF data) was stable for 10th and 12th grade males; use among eighth grade males decreased significantly (P < .01). Since 1991, data from the 3 national surveys indicate an increase in lifetime anabolic steroid use among adolescent females, although only 1 of these increases is statistically significant. Furthermore, past year use of steroids (MTF data) increased for females in the 8th (P < .05), 10th (P < .05), and 12th (ns) grades. CONCLUSIONS: A long-term comparison of anabolic steroid use (from 1989-1996) indicates that use among adolescent males and females has decreased significantly (P < .05). However, for females the low point in lifetime steroid use was reached in 1991, with subsequent significant (P < .05) increases in use being reported in several national data sets. For adolescent males, after declining sharply between 1989 and 1991, steroid use has generally been stable since 1991. Moreover, based on the 1995 estimates of high school students and Youth Risk and Behavior Surveillance System data, approximately 375,000 adolescent males and 175,000 adolescent females in public and private schools in the United States used anabolic steroids at least once in their lives. These results suggest that prevention, intervention, and regulatory efforts to reduce steroid use at the local, state, and national levels should be reassessed, especially those efforts that focus on adolescent female steroid use. PMID- 9412595 TI - Risks for bacteremia and urinary tract infections in young febrile children with bronchiolitis. AB - OBJECTIVE: To compare the risks for bacteremia and urinary tract injections (UTI) in young febrile children with and without bronchiolitis. DESIGN: A prospective cohort study. SETTING: The emergency departments of 3 pediatric referral hospitals. PATIENTS: A convenience sample of 432 previously healthy febrile patients aged 24 months or younger. Patients were divided into groups, based on the presence (n = 163, bronchiolitis group) or absence (n = 269, control group) of wheezing and/or retractions on examination. Blood cultures were obtained from all patients, and urine cultures were obtained from female patients, and male patients aged 6 months or younger. Chest radiographs were obtained on patients with lower respiratory tract signs, and those with lobar pneumonias were excluded (7 wheezing and 8 nonwheezing patients), leaving 156 patients with bronchiolitis and 261 control patients. OUTCOME MEASURES: Growth of any bacterial pathogens from the blood or 10(4) colony-forming units per milliliter or more from the urine. RESULTS: None of the 156 patients with bronchiolitis had bacteremia (95% confidence interval, 0%-1.9%) vs 2.7% of the 261 controls (95% confidence interval, 1.1%-5.4%; P = .049); 1.9% of the patients with bronchiolitis had UTI vs 13.6% of the controls (odds ratio, 0.12; 95% confidence interval, 0.02-0.55; P = .001). None of the subset of patients with bronchiolitis aged 2 months or younger (n = 36) had bacteremia or UTI; however, there were not enough of these younger patients to make statistically conclusive comparisons. CONCLUSIONS: Previously healthy febrile children aged 24 months or younger with bronchiolitis are unlikely to have bacteremia or UTI. Therefore, routine cultures of the blood and urine in these patients are unnecessary. More data are needed regarding the subset of febrile infants aged 2 months or younger with bronchiolitis. PMID- 9412596 TI - Persistently increased injury mortality rates in high-risk young children. AB - OBJECTIVE: To study trends in injury mortality for low- and high-risk young children. DESIGN AND METHODS: For Tennessee children 0 to 4 years of age, we used birth certificates to obtain data on maternal education, age, and parity; these risk factors were used to classify children into low- and high-risk groups. The outcome was death from injury, as determined from linked death certificates. Between 1978 and 1995, injury mortality rates were calculated for six 3-year periods for low- and high-risk children. RESULTS: There were 1.5 million children 0 to 4 years of age who contributed 4.9 million child-years. The high-risk group contributed 28% of all child-years. There were 673 injury deaths in the high-risk group, 48.9 deaths per 100,000 child-years, and 586 deaths in the low-risk group, 16.8 deaths per 100,000 child-years. The injury mortality rate for low-risk children decreased from 20.7 to 15.7 per 100,000 child-years between the 1978 1980 and 1981-1983 periods; thereafter it remained relatively stable. For high risk children, the injury mortality rate decreased from 50.9 to 43.5 per 100,000 between the 1978-1980 and 1981-1983 periods, remained mostly unchanged through 1992, and then increased sharply in the 1993-1995 period to 64.1 per 100,000 child-years. The disparity between high- and low-risk children widened from 29.3 (95% confidence interval, 25.1-33.5) excess deaths per 100,000 for 1978 through 1991 to 46.9 (95% confidence interval, 35.9-57.9) in 1993 through 1995. CONCLUSIONS: In Tennessee, maternal education, age, and parity consistently identified a population of children at increased risk of injury mortality. For these high-risk children, there has been no substantial reduction in injury mortality in high-risk young children during the last 18 years. PMID- 9412597 TI - A clinic system to improve preschool vaccinations in a low socioeconomic status population. AB - OBJECTIVE: To determine if a clinic system to assess and vaccinate preschool-age children at every clinic visit can improve vaccination rates. DESIGN: A nonequivalent control group design contrasting an intervention clinic with a comparison clinic. SETTING: Two urban St Paul, Minn, clinics. The intervention clinic is a family practice residency clinic, and the comparison, clinic is a community health center clinic. PATIENTS: Primarily a low socioeconomic status white population. INTERVENTIONS: A clinic-wide system to identify and vaccinate children at all clinic visits. Appointment personnel, medical assistants, and physicians all had roles in the intervention protocol. MAIN OUTCOME MEASURES: Percentage of children at the 2 clinics who were up-to-date for a primary vaccine series at age 24 months and also at the end of the study collection periods, preintervention and postintervention. RESULTS: The intervention clinic improved the percentage of children up-to-date for a primary vaccine series at age 24 months from 42% to 56% (P = .02), while the percentage at the comparison clinic did not change significantly (P = .81). Similarly, the intervention clinic improved the percentage of children up-to-date for age at the end of the study periods from 49% preintervention to 63% postintervention (P = .02), while the percentage at the comparison clinic did not improve significantly (P = .45). The system was especially useful for children with few visits to the intervention clinic. CONCLUSIONS: Although the intervention clinic resulted in a substantial improvement in vaccination rates for preschool-age children, rates remained well below national goals. A combination of clinic, community, and national initiatives may be needed to ensure appropriate vaccination rates for this challenging patient population. PMID- 9412598 TI - Cerebrospinal fluid findings in children with Lyme disease-associated facial nerve palsy. AB - OBJECTIVE: To determine the relative frequency of abnormal cerebrospinal fluid (CSF) findings in children with Lyme disease-associated facial nerve palsy. DESIGN: A clinical series. A prospective evaluation was undertaken of the condition of children seen between 1988 and 1996 at a single medical center in a Lyme disease endemic area. PATIENTS: Forty children (24 boys and 16 girls, aged 3 19 years) with new onset facial nerve palsy who met the Centers for Disease Control and Prevention case definition of Lyme disease. INTERVENTIONS: Neurologic examinations. Cerebrospinal fluid analysis. MAIN OUTCOME MEASURES: Rates of abnormal CSF findings: white blood cell count, protein level, and Borrelia burgdorferi-specific CSF assays. RESULTS: Cerebrospinal fluid white blood cell count, protein level, or both were abnormal in 27 (68%) of the children. Thirty six (90%) of the 40 children had a CSF abnormality consistent with central nervous system infection or immune involvement by B burgdorferi. Of the 22 children with CSF pleocytosis, only 7 (32%) had headache and none had meningeal signs. CONCLUSIONS: Most children with Lyme disease-associated facial nerve palsy have CSF abnormalities. Our studies indicate that, in endemic areas, facial nerve palsy in children may be a marker of Lyme disease and occult meningitis. When Lyme disease is suspected, CSF should be examined; in some cases, it may be helpful to expand beyond routine CSF studies to look at a battery of B burgdorferi-specific assays. PMID- 9412599 TI - Effect of the vaccines for children program on inner-city neighborhood physicians. AB - OBJECTIVE: To determine the probable effect of the Vaccines for Children (VFC) program on immunization coverage. DESIGN: Preintervention and postintervention study design, with data collected before and after enrollment in the VFC program. SETTING: Twenty-three inner-city neighborhood physicians' offices in New York City. PARTICIPANTS: In 1993, 30 physicians were randomly selected from 8 neighborhoods with the highest proportions of Medicaid-eligible individuals in New York City. In 1995-1996, the 30 physicians were contacted again. Twenty-three agreed to an interview and medical record review. Within each office, the medical records of children aged 3 to 35 months, with at least 3 visits in a 3-month or longer period, were randomly selected. Medical record reviews were conducted for 173 eligible children in 1993 and 528 in 1995-1996. INTERVENTIONS: The VFC program was implemented in October 1994. The administration fee increased from $2 to $17.85; physicians received vaccines free. MAIN OUTCOME MEASURES: Up-to-date status for immunizations and lead and tuberculosis screening; percentage of visits that are missed opportunities to immunize; and percentage of visits that were well-child visits. Up-to-date status, missed opportunities to immunize, and well-child visits were compared across time using chi 2 analysis, corrected for the use of cluster sampling. RESULTS: Up-to-date status changed significantly before and alter enrollment in the VFC program (P < .05) for all immunizations and for lead and tuberculosis screening. For the diphtheria toxoid, tetanus toxoid, and pertussis vaccine, oral poliovirus vaccine, and measles, mumps, and rubella vaccine combined, coverage increased from 17.9% to 42.2%, up by 24.3 percentage points (P < .05). Missed opportunities to immunize did not change, but well-child visits increased from 15.0% to 21.6% (P < .05). Physicians generally attributed performance improvements to the VFC program and not to other competing hypotheses. CONCLUSIONS: The VFC program seems to be responsible for an increase in immunization rates among these physicians. PMID- 9412600 TI - Children with in utero cocaine exposure do not differ from control subjects on intelligence testing. AB - OBJECTIVE: To determine if in utero cocaine exposure affects IQ scores in children at age 4 years. DESIGN: A prospective, longitudinal evaluation by blinded examiners of the IQ scores of cocaine-exposed and control children of low socioeconomic status who have been observed since birth. SETTING: A study center in an inner-city hospital. PARTICIPANTS: One hundred one children with in utero cocaine exposure and 118 control children, all of whom were 34 weeks' gestational age or older and nonasphyxiated at birth. MAIN OUTCOME MEASURE: Intelligence quotient scores on a standardized intelligence test, the Wechsler preschool and Primary Scale of Intelligence--Revised. RESULTS: Seventy-one cocaine-exposed and 78 control children were administered the Wechsler Preschool and Primary Scale of Intelligence--Revised. Maternal, natal, and 30-month characteristics of the children tested did not differ from those not tested. Groups did not differ on mean Performance (83.2 vs 87.0), Verbal (79.0 vs 80.8), or Full Scale (79.0 vs 81.9) IQ scores (all P > or = .10 [values for cocaine-exposed children given first]). None of these 3 scores was associated with cocaine exposure in multivariate linear regressions. Although cocaine-exposed and control groups did not differ in outcome, 93% of cocaine-exposed and 96% of control children had Full Scale IQ scores below 100, the mean IQ score for the test. CONCLUSIONS: In an inner-city cohort, IQ scores did not differ between cocaine-exposed and control children. However, both groups performed poorly. PMID- 9412601 TI - New use of anticonvulsant medications among children enrolled in the Tennessee Medicaid Program. AB - OBJECTIVE: To describe the new use of anticonvulsant medications among children enrolled in the Tennessee Medicaid program. DESIGN: A retrospective cohort study. PATIENTS: New users of anticonvulsant medications in 1992 were identified from the 206,098 children (aged 0-18 years) enrolled continuously for 12 months in the Aid to Families With Dependent Children program or foster care program of Tennessee Medicaid. MAIN OUTCOME MEASURES: New users were categorized according to the diagnosis codes of health care encounters occurring 90 days before to 90 days after the first anticonvulsant prescription was filled as having diagnoses consistent with (1) epilepsy or convulsions, (2) neonatal seizures, (3) central nervous system disease, (4) no epilepsy diagnoses but diagnoses for which anticonvulsants might appropriately be used (jaundice, headaches, or psychiatric disorders), or (5) no diagnoses for which an anticonvulsant might appropriately be used. The children in each group were described according to sociodemographic variables, with logistic regression used to analyze variations in the subsequent filling of anticonvulsant prescriptions. RESULTS: Of 647 children continuously enrolled in the Tennessee Medicaid program who were new anticonvulsant users in 1992, 58% had at least 1 health care encounter coded as epilepsy or convulsions, 2% had a diagnosis of neonatal seizures, 8% had central nervous system diagnoses, 16% had specific nonepilepsy diagnoses (jaundice, headache, or psychiatric diagnoses), and 16% had no diagnoses for which anticonvulsants might appropriately be prescribed. For children with epilepsy diagnoses, white race (P = .002) and undergoing tests (P < .001) were independent predictors of a child filling 6 or more prescriptions in the year following the first prescription CONCLUSIONS: A large proportion of new users of anticonvulsants among children enrolled in the Tennessee Medicaid program received these medications for indications other than epilepsy. For children with epilepsy diagnoses, there was considerable variation in the subsequent filling of prescriptions. Further analysis of these variations in practice will allow for the development of policies that will maximize benefit for children who need anticonvulsant therapy, while diminishing unnecessary exposure to potentially toxic drugs for children who do not. PMID- 9412602 TI - Transition to a computer-based record using scannable, structured encounter forms. AB - OBJECTIVE: To evaluate the quality of documentation and user satisfaction with a structured documentation system for pediatric health maintenance encounters, using scanned paper-based forms to generate an electronic medical record. DESIGN: (1) A retrospective medical record review comparing 16 structured (ST) records with 16 contemporaneously created unstructured records, (2) a questionnaire evaluation of user satisfaction, and (3) an electronic records review of patients seen 1 year following the full implementation of the system to evaluate persistence of the effect. SETTING: The Yale-New Haven Hospital Pediatric Primary Care Center, New Haven, Conn, an inner-city clinic in an academic center. PARTICIPANTS: (1) A random sample of 16 health maintenance records completed by first- and second-year residents in February 1996 matched for patient's age and provider training level with 16 contemporaneously documented visits, (2) 16 of 18 pediatric level 1 residents and 14 of 16 pediatric level 2 residents who completed questionnaires, and (3) all electronic records of health maintenance visits during February 1997. MAIN OUTCOME MEASURES: The number of data elements documented and the percentage of records that record specific components of the health maintenance encounter. User satisfaction was specified on a Likert scale. RESULTS: Overall, residents in the ST records group documented more data elements per visit than did those in the unstructured records group. The number of developmental items documented was 11.5 per visit in the ST records group and 4.8 per visit in the unstructured records group (P = .004). Likewise, anticipatory guidance was more thoroughly documented in the ST records group--8.3 items per visit vs 2.5 items per visit (P < .001). Ninety percent of the users preferred the ST records. One year after the adoption of the ST recording system, high levels of thoroughness persisted. CONCLUSIONS: Structured, scannable encounter forms can facilitate documentation of patient care and are well accepted by users. They can provide an effective mechanism to ease the transition to a computer-based patient record. PMID- 9412603 TI - Pediatric residents' telephone triage experience. Relevant to general pediatric practice? AB - OBJECTIVES: To describe a pediatric resident telephone triage system in a tertiary hospital and to determine its relevance to telephone experience in general pediatric practice DESIGN: An analysis of 514 telephone calls from parents of continuity clinic patients made to pediatric residents. The evaluation included: chief complaint, disposition of call, age of patient, and level of training of the resident answering the call. A comparison was made with published information about a private-practice telephone triage system. SETTING: Pediatric continuity clinic, Medical College of Georgia, Augusta. PATIENTS: Children registered in the pediatric resident continuity clinic at the Medical College of Georgia. RESULTS: The 13 most frequent reasons for calling were some of the most common problems seen in pediatric practice. The disposition of calls was as follows: 272 (53%) were given telephone advice alone, 119 (23%) were offered an appointment for the next day, and 123 (24%) were advised to come to the emergency department immediately. Disposition did not vary with residency level. Both chief complaints and disposition of calls were similar to those reported in a private practice nurse triage system. CONCLUSIONS: Answering telephone calls in a residency telephone triage system, when combined with a curriculum that includes next-day monitoring, feedback from a preceptor, and seminar discussions focused on telephone management situations, is a valuable training experience and is relevant for residents going into private pediatric practice. PMID- 9412604 TI - Radiological case of the month. Persistent left fifth aortic arch in a child without congenital heart disease. PMID- 9412605 TI - Radiological case of the month. Choroidal hemangioma. PMID- 9412606 TI - Picture of the month. Albright hereditary osteodystrophy. PMID- 9412607 TI - Pathological case of the month. Abetalipoproteinemia (Bassen-Kornzweig syndrome). PMID- 9412608 TI - Pathological case of the month. Fetal brain disruption sequence. PMID- 9412609 TI - Tobacco and children: an economic evaluation of the medical effects of parental smoking. PMID- 9412610 TI - Antecedents of cerebral palsy in a multicenter trial of indomethacin for intraventricular hemorrhage. PMID- 9412611 TI - Lubeluzole treatment of acute ischemic stroke. The US and Canadian Lubeluzole Ischemic Stroke Study Group. AB - BACKGROUND AND PURPOSE: Lubeluzole is a novel benzothiazole compound that has shown neuroprotective activity in preclinical models of ischemic stroke. The present multicenter, double-blind, placebo-controlled study was conducted to assess the efficacy and safety of lubeluzole in the treatment of ischemic stroke. METHODS: Seven hundred twenty-one patients with clinical symptoms of acute ischemic stroke were randomized to receive either lubeluzole (7.5 mg over 1 hour, followed by a continuous daily infusion of 10 mg for up to 5 days) or placebo. Treatment was initiated within 6 hours of symptom onset. Mortality at 12 weeks was the primary efficacy end point. Secondary efficacy end points included neurological recovery (based on the National Institutes of Health Stroke Scale [NIHSS]), functional status (based on the Barthel Index), and level of disability (based on the Rankin Scale). Safety assessments included standard and continuous electrocardiographic monitoring, physical examination, measurements of vital signs, clinical laboratory evaluation, and adverse events reports. RESULTS: The overall mortality rate at 12 weeks for lubeluzole-treated patients was 20.7% compared to 25.2% for placebo-treated patients (NS). Controlling for relevant covariates, the degree of neurological recovery (NIHSS) at week 12 significantly favored lubeluzole over placebo (P = .033). Lubeluzole treatment similarly resulted in significantly greater improvements in functional status (Barthel Index) (P = .038) and overall disability (Rankin Scale) (P = .034) after 12 weeks. A global test statistic confirmed that lubeluzole-treated patients had a more favorable clinical outcome at 12 weeks (P = .041). The safety profile of lubeluzole resembled that of placebo. CONCLUSIONS: Treatment with lubeluzole within 6 hours of the onset of ischemic stroke had a nonsignificant effect on mortality and resulted in improved clinical outcome compared with placebo, with no safety concerns. PMID- 9412612 TI - Treatment of acute ischemic stroke with piracetam. Members of the Piracetam in Acute Stroke Study (PASS) Group. AB - BACKGROUND AND PURPOSE: Piracetam, a nootropic agent with neuroprotective properties, has been reported in pilot studies to increase compromised regional cerebral blood flow in patients with acute stroke and, given soon after onset, to improve clinical outcome. We performed a multicenter, randomized, double-blind trial to test whether piracetam conferred benefit when given within 12 hours of the onset of acute ischemic stroke to a large group of patients. METHODS: Patients received placebo or 12 g piracetam as an initial intravenous bolus, 12 g daily for 4 weeks and 4.8 g daily for 8 weeks. The primary end point was neurologic outcome after 4 weeks as assessed by the Orgogozo scale. Functional status at 12 weeks as measured by the Barthel Index was the major secondary outcome. CT scan was performed within 24 hours of the onset of stroke but not necessarily before treatment. Analyses based on the intention to treat were performed in all randomized patients (n = 927) and in an "early treatment" population specified in the protocol as treatment within 6 hours of the onset of stroke but subsequently redefined as less than 7 hours after onset (n = 452). RESULTS: In the total population, outcome was similar with both treatments (the mean Orgogozo scale after 4 weeks: piracetam 57.7, placebo 57.6; the mean Barthel Index after 12 weeks: piracetam 55.8, placebo 53.1). Mortality at 12 weeks was 23.9% (111/464) in the piracetam group and 19.2% (89/463) in the placebo group (relative risk 1.24, 95% confidence interval, 0.97 to 1.59; P = .15). Deaths were fewer in the piracetam group in those patients in the intention-to-treat population admitted with primary hemorrhagic stroke. Post hoc analyses in the early treatment subgroup showed differences favoring piracetam relative to placebo in mean Orgogozo scale scores after 4 weeks (piracetam 60.4, placebo 54.9; P = .07) and Barthel Index scores at 12 weeks (piracetam 58.6, placebo 49.4; P = .02). Additional analyses within this subgroup, confined to 360 patients with moderate and severe stroke (initial Orgogozo scale score < 55), showed significant improvement on piracetam in both outcomes (P < .02). CONCLUSIONS: Piracetam did not influence outcome when given within 12 hours of the onset of acute ischemic stroke. Post hoc analyses suggest that piracetam may confer benefit when given within 7 hours of onset, particularly in patients with stroke of moderate and severe degree. A randomized, placebo-controlled, multicenter study, the Piracetam Acute Stroke Study II (PASS II) will soon begin. PMID- 9412613 TI - Sex dependency of cerebrovascular CO2 reactivity in normal subjects. AB - BACKGROUND AND PURPOSE: Cerebrovascular CO2 reactivity can be assessed easily and reliably by transcranial Doppler sonography. The objectives of the present study were to evaluate sex differences in cerebral CO2 reactivity and to specify the relation between CO2 and cerebral blood flow velocity. METHODS: CO2 reactivity of the circulation of both middle cerebral arteries was measured by bilateral transcranial Doppler sonography in 60 healthy volunteers (30 men, 30 women) aged 21 to 58 years. End-tidal carbon dioxide tensions (PETCO2) were elevated with the use of carbogene gas (95% O2, 5% CO2). In each subject the mean blood flow velocity (Vmean) was plotted as a function of PETCO2. RESULTS: The best-fit curves for the relation of Vmean/PETCO2 were exponential functions, with the following basic equation: Vmean (cm/s) = aebx, where a is a theoretical quantity representing Vmean at a PCO2 of 0 mm Hg, b is the relative slope of the curve (slope divided by the value of the function) corresponding to the definition of reactivity, and x is the PETCO2 (mm Hg). The mean value of b was 0.037 +/- 0.008 in women and 0.030 +/- 0.010 in men. ANOVA demonstrated a significant difference between men and women (P < .001). CONCLUSIONS: This study demonstrates a highly significant sex-related difference in CO2-induced cerebral vasomotor reactivity. The relation between altered carbon dioxide tensions and blood flow velocities of both middle cerebral arteries in 60 healthy volunteers was found to be exponential. PMID- 9412614 TI - Cardiopulmonary and arterial baroreflex-mediated control of forearm vasomotor tone is impaired after acute stroke. AB - BACKGROUND AND PURPOSE: Elevated blood pressure (BP) levels are well recognized after acute stroke and are associated with increased BP variability. The underlying mechanisms producing such changes are unclear but may include abnormalities of baroreceptor-mediated control of heart rate and vasomotor tone. Lower body negative pressure (LBNP) can be used to assess the integrity of "low pressure" cardiopulmonary and "high-pressure" arterial baroreceptor-derived responses by inducing nonhypotensive and hypotensive stimuli. METHODS: Cardiovascular responses, including BP, heart rate, forearm blood flow, and forearm vascular resistance, to nonhypotensive and hypotensive LBNP were assessed in 13 consecutive stroke patients. Patients were studied within 72 hours of stroke (acute) and again at 10 to 14 days (subacute) and were compared with 13 control subjects individually matched for age, sex, and BP. RESULTS: At an LBNP of -10 mm Hg, BP was unchanged in all groups, but a significant increase in forearm vascular resistance occurred only in the control group (11 U [interquartile range, 7 to 15]; P < .05) compared with stroke patients in the acute (9 U [3 to 14]; P = NS) or subacute phases (7 U [2 to 12]; P = NS). After LBNP at -40 mm Hg, the reductions in systolic BP levels were similar in all groups (control: -9 mm Hg [-16 to -3]; acute stroke: -9 mm Hg [-22 to 3]; subacute stroke: -7 mm Hg [-35 to 20]), as was the associated increase in heart rate (control: 8 bpm ([4 to 11]; acute stroke: 6 bpm ([1 to 12]; subacute stroke: 9 bpm [2 to 19]). However, forearm vascular resistance increased significantly only in control subjects (20 U [9 to 30]; P < .01). CONCLUSIONS: The present study has identified abnormal vasomotor responses to LBNP after acute stroke, with an increase in FVR only being observed in control subjects in response to nonhypotensive and similar hypotensive levels of LBNP. In acute stroke patients, the stimulus of hypotensive LBNP appears to be compensated by an increase in cardiac output since there appears to be no increase in peripheral vascular resistance, unlike the changes seen in control subjects. However, the exact mechanisms for these changes are still unclear and are the subject of further study. PMID- 9412615 TI - Cerebral venous thrombosis and anticardiolipin antibodies. AB - BACKGROUND AND PURPOSE: The association of cerebral venous thrombosis (CVT) with a variety of pathological states is well established. However, there are only rare isolated reports of CVT associated with anticardiolipin antibodies (aCL). METHODS: To clarify the clinical and neuradiological features as well as outcome of patients with CVT associated with aCL, we reviewed the records of all patients with CVT evaluated at our institution between 1989 and 1996 (retrospective and prospective) and systematically reviewed the pertinent literature. RESULTS: We identified 8 aCL+ and 7 aCL- patients with CVT. No patients with lupus anticoagulant (LA) were identified. The mean age was 23 +/- 11.01 (range, < 1 to 36) years in the aCL+ and 38 +/- 9.30 (range, 25 to 54) years in the aCL- patients (P = .016). Six of 8 aCL+ and 5 of 7 aCL- patients were women. The dural sinuses were involved in all aCL+ and in 6 of 7 aCL- patients, while deep venous system thrombosis occurred in 5 of 8 (63%) aCL+ and 1 of 7 (14%) aCL- patients. In the aCL+ patients CVT was associated with puerperium or oral contraceptive use (n = 6), and sickle cell trait (n = 1), and in the aCL- patients CVT was associated with systemic lupus erythromatosus (n = 1), myelodysplasti syndrome (n = 1), colonic cancer (n = 1), oral contraceptive use or puerperium (n = 3), and dehydration (n = 1). Seven aCL+ patients received either intrasinus urokinase or intravenous heparin sulfate, and 1 received aspirin. Four aCL+ patients developed new onset or worsening of preexisting migraine, 2 developed recurrent peripheral venous thrombosis, and 1 went on to have intracranial hypertension. Twenty additional patients with CVT associated with antiphospholipid antibodies (aPL) were found reported in the literature. The overall mean age was 36 +/- 11.6 (range, 21 to 62) years, and 14 (70%) were women. LA was present in 11 of 18 tested, aCL in 7 (35%), LA and aCL in 1, and the type of aPL was not reported in 3. The mean age for the aCL+ only group was 28 years and for the LA+ (with or without aCL+) was 34 years. Only 1 patient, whose aPL type was not specified, had thrombosis of the deep venous system in addition to involvement of the dural sinuses. CONCLUSIONS: Our series and review suggest that aCL may be an important factor contributing to development of CVT even in the presence of other potential etiologies or risk factors. Onset of aCL+ CVT occurs at a relative young age and with relatively more extensive superficial and deep cerebral venous system involvement than aCL- CVT. PMID- 9412616 TI - Predisposing factors to enlargement of spontaneous intracerebral hematoma. AB - BACKGROUND AND PURPOSE: Enlargement of intracerebral hemorrhage is a major cause of clinical deterioration. Identification of factors that predispose to hematoma enlargement is important in managing patients. METHODS: We selected 186 patients (71 women and 115 men; mean age, 64.8 +/- 12.5 years) with spontaneous intracerebral hemorrhage who had undergone an initial CT within 24 hours and a second scan within 120 hours of symptom onset. We compared patients with (n = 41) and without (n = 145) hematoma enlargement according to clinical characteristics and laboratory data. RESULTS: By multiple logistic regression analysis (n = 139), interaction of long interval (> 6 hours) from onset to first CT and small hematoma (< 25 cm3) strongly reduced risk of enlargement. The analysis also demonstrated that the following factors independently predisposed to enlargement: history of brain infarction; liver disease; interaction of fasting plasma glucose > or = 141 mg/dL and systolic blood pressure on admission > or = 200 mm Hg; and interaction of glycosylated hemoglobin A1c > or = 5.1% and systolic blood pressure on admission > or = 200 mm Hg. CONCLUSIONS: A patient examined > 6 hours after ictus who has a hematoma volume < 25 cm3 is unlikely to experience further hematoma growth. Prevention of brain infarction and premorbid management of liver disease may serve to lower the risk of hematoma enlargement. Although it remains controversial whether antihypertensive drugs should be used in the acute phase of intracerebral hemorrhage, poorly controlled diabetics with high systolic blood pressure (> or = 200 mm Hg) on admission also were at high risk of hematoma enlargement. PMID- 9412617 TI - Surgical closure of patent foramen ovale in cryptogenic stroke patients. AB - BACKGROUND AND PURPOSE: Patents foramen ovale (PFO) is associated with stroke of unknown etiology or cryptogenic stroke. However, optimal treatment to prevent recurrence in cryptogenic stroke patients with PFO is not clearly defined. Since PFO represents a surgically repairable lesion, interest in closing it is high. This report reviews our experience with cryptogenic stroke patients with PFO who underwent surgical PFO closure and were followed for recurrence of neurological events. METHODS: We followed 28 cryptogenic stroke patients (17 men, 11 women; mean age, 41 +/- 13 years) with transesophageal echocardiography-defined PFO who had undergone PFO closure by open thoracatomy. All patients selected for surgery refused, could not take, or failed warfarin therapy. They were followed by physician visits and telephone interviews. RESULTS: There were no surgical complications. With a mean follow-up of 19 months, four patients experienced neurological event recurrence, one stroke, and three transient ischemic attacks. Kaplan-Meier survival analysis demonstrated that the actuarial rate of recurrence was 19.5% (95% confidence limit 2.2-36.8%) at 13 months of follow-up. None of the 17 patients (0%) younger than 45 years suffered a recurrence, whereas four of 11 patients (35%) aged 45 or older experienced a recurrence of neurological event (P < .02). Using a proportional hazards regression model, the increase in relative risk with increasing age was 2.76 per 10 years (95% confidence interval 1.07 to 7.16). CONCLUSIONS: Although PFO is easily repairable in patients with crytogenic stroke, its closure does not consistently prevent recurrence of ischemic events. The recurrence appears to occur more frequently in older cryptogenic stroke patients. PMID- 9412618 TI - Warfarin use among patients with atrial fibrillation. AB - BACKGROUND AND PURPOSE: Warfarin reduces the rate of stroke among patients with atrial fibrillation. We sought to determine warfarin use within a population sample of elderly patients with atrial fibrillation. METHODS: The Connecticut Peer Review Organization conducted a chart review of Medicare patients aged > or = 65 years with a history of atrial fibrillation before a hospitalization during the first 6 months of 1994. RESULTS: Among 488 patients (308 women; 457 white; 173 aged > or = 85 years), 38% (184/488) had a relative contraindication to anticoagulation (history of bleeding, dementia, alcohol use, falls, cancer, or the need for nonsteroidal anti-inflammatory drugs). Among the remaining patients (with known atrial fibrillation, but without a contraindication), only 38% (117/304) had been prescribed warfarin. Of those not prescribed warfarin, 63% (117/187) were also not taking aspirin. There were 272 patients with at least one additional vascular risk factor and no contraindication to anticoagulation. Among these patients at moderate to high risk for stroke, anticoagulation had been prescribed in 40% (109/272). Overal, among those not prescribed warfarin, 58% (95/163) were not taking aspirin. Patients admitted with a stroke were more likely to be significantly underanticoagulated (with international normalized ratio < 1.5) (43.5% versus 20.9% for those without stroke; P < .005). Anticoagulation was most effective for those with an international normalized ratio > or = 2.0. CONCLUSIONS: Warfarin anticoagulation with atrial fibrillation, even among "ideal" candidates, appears dramatically underutilized. In addition, among those prescribed warfarin, patients are often undertreated. Increased warfarin use among patients with atrial fibrillation represents an excellent opportunity for stroke prevention in the elderly. PMID- 9412619 TI - Long-term oral anticoagulation reduces bone mass in patients with previous hemispheric infarction and nonrheumatic atrial fibrillation. AB - BACKGROUND AND PURPOSE: Vitamin K is an essential factor for synthesis of plasma clotting proteins and for site-specific carboxylation of bone Gla protein and other bone matrix proteins. Low vitamin K has been associated with reduced bone mineral density. Warfarin therapy, which inhibits vitamin K-dependent blood clotting, has been demonstrated to reduce the risk of stroke in nonrheumatic atrial fibrillation. We evaluated vitamin K and bone mineral density in nonrheumatic atrial fibrillation patients who had long-term warfarin therapy after an ischemic stroke. METHODS: Sera were collected from 64 patients with non rheumatic atrial fibrillation and ischemic stroke who had been treated with warfarin, 63 stroke patients without warfarin, and 39 control subjects. All stroke patients in both groups had hemiplegia. Sera were assayed for vitamins K1 and K2, bone Gla protein, and 25-hydroxyvitamin D. Bone mineral density was determined in both second metacarpals. RESULTS: Serum vitamin K1 concentrations (ng/mL) were lower in treated patients (.234 +/- .177 ng/mL) than in untreated patients (.329 +/- .284) or controls (.553 +/- .164). Bone Gla protein was lower in treated patients' sera (1.241 +/- .799 ng/mL) than in untreated patients (4.476 +/- 3.226). Concentrations of 25-hydroxyvitamin D were lower in both patient groups. Bone mineral density was lower on both sides in treated patients than in untreated patients (P < .0001). Vitamin K1 and bone Gla protein were significantly related to bone mineral density bilaterally in treated but not in untreated patients. CONCLUSIONS: Bone mineral density was significantly lower in stroke patients with long-term warfarin treatment than in untreated patients. Both warfarin-induced reduction in vitamin K function and lowered vitamin K1 concentrations are probable causes of this osteopenia. PMID- 9412620 TI - Quality of life of stroke in younger individuals. Utility assessment in patients with arteriovenous malformations. AB - BACKGROUND AND PURPOSE: Patients with arteriovenous malformations are younger individuals who are at risk of a stroke or have experienced one. Our objective was to determine these patients' perceptions of quality of life with a stroke by eliciting utility values. METHODS: Utility values were obtained with the standard gamble technique. Utilities are a holistic, quality of life measure between 0 and 1. We evaluated the patients' current health state and written descriptions of major and minor stroke. RESULTS: Thirty-one consecutive outpatients participated. The mean age was 37 years (range, 18 to 57 years). Approximately 65% had suffered a stroke, of which 55% were major. Approximately 61% had a persistent deficit. The mean utilities were 0.45 for major stroke (95% confidence interval [CI], 0.33 to 0.56; range, 0.00 to 1.00), 0.81 for minor stroke (95% CI, 0.75 to 0.88; range, 0.30 to 1.00), and 0.81 for current health (95% CI, 0.73 to 0.89; range, 0.01 to 1.00). Subgroup analyses by demographic and clinical characteristics showed no significant differences. However, in both those patients who had never had a stroke and those who had survived a major stroke, values for the major stroke scenario were clustered at the two extremes. Among those with a current deficit, 79% preferred their own health state to that of the stroke scenario that was similar in severity. CONCLUSIONS: Younger people who have had a stroke or are at risk demonstrate wide variations in their perception of quality of life. Furthermore, patients tend to be more risk averse with their own lives than with theoretical scenarios. We suggest tailoring medical decision making to individual preferences. PMID- 9412621 TI - Causes and mechanisms of cerebellar infarction in young patients. AB - BACKGROUND AND PURPOSE: The incidence of cerebellar infarction in a series of patients with stroke is approximately 1.5%. The average patient age in most reported series is 62 years. The most common etiologies in this age group are atherosclerosis and cardiac embolism. The aim of this study was to determine the causes and mechanisms of cerebellar infarction in patients younger than 40 years. METHODS: We analyzed retrospectively the clinical and radiological data from 21 men and 16 women with cerebellar infarction admitted to our hospital from January 1986 to December 1996. The patients had been studied extensively to determine the etiology of the cerebellar infarction. RESULTS: In the 37 patients (mean age, 30 years), 29 infarcts were limited to one territory (15 in the posteroinferior cerebellar artery [PICA]; 14 in the superior cerebellar artery); 8 had nonterritorial infarctions. The most common stroke mechanisms in each territory were as follows: PICA: nonatherosclerotic vasculopathic (67%), cardioembolic (20%), and hematologic and cryptogenic (each 7%); superior cerebellar artery: cardioembolic (42%), cryptogenic (31%), migrainous (21%), and nonatherosclerotic vasculopathic and hematologic (each 7%); and mixed territory: nonatherosclerotic vasculopathic (50%), cryptogenic (25%), cardioembolic (12%), and hematologic (12%). CONCLUSIONS: The most common mechanism of cerebellar infarctions was arterial occlusion as a result of intracranial vertebral artery dissection (40%), mainly with PICA involvement. Embolism from a cardiac source resulted primarily from patent foramen ovale and rheumatic valvular disease. Hematologic disturbances and migraine were responsible for a few cases. PMID- 9412622 TI - Outcome in 43 patients with distal anterior cerebral artery aneurysms. AB - BACKGROUND AND PURPOSE: The aim of this retrospective multicenter study was to evaluate the outcome of distal anterior cerebral artery (DACA) aneurysms and to determine the incidence, causes, and consequences of unfavorable outcomes. METHODS: 43 patients with 50 DACA aneurysms (27 females and 16 males, mean age 49 years) were studied retrospectively. Forty-four DACA aneurysms were treated surgically (83% with an interhemispheric approach), and 2 were embolized. At postoperative day 10, all patients underwent routine angiography. The outcome at 6 to 12 months was scored according to the Glasgow Outcome Scale (GOS). RESULTS: 35 DACA aneurysms were ruptured. Among the 26 "good"-grade patients (Hunt and Hess grades I through III), 18 (69.2%) were in GOS 1, 2 in GOS 2 (7.7%), 2 in GOS 3 (7.7%), and 4 in GOS 5 (15.4%); among the 9 "poor"-grade patients (Hunt and Hess grades IV and V), 1 (11.1%) was in GOS 1, 2 in GOS 2 (22.2%), 2 in GOS 3 (22.2%), and 4 in GOS 5 (44.5%). The initial intracerebral hemorrhage (ICH) (40%) induced neurological aftereffects in 8 patients. An operative rupture occurred in 40%, with a temporary occlusion in 28.6% that was responsible for mediocre results in 3 patients (8.7%). A postoperative thrombosis was observed in 4 patients (11.4%) and an aneurysmal remnant in 1 (2.8%). Ten DACA unruptured aneurysms were clipped without operative rupture or thrombosis. CONCLUSIONS: The authors suggest that the proportion of ruptured DACA aneurysms evolving to a GOS 1 or 2 was similar to that of aneurysms found in other locations, after early surgery. Endovascular treatment should be considered in the management of uncommon ruptured fusiform DACA aneurysms. PMID- 9412623 TI - Apolipoprotein E epsilon 2 allele and risk of stroke in the older population. AB - BACKGROUND AND PURPOSE: There is evidence for a role of apolipoprotein E (apoE) in atherosclerosis. Coronary heart disease morbidity is higher in persons carrying an epsilon 4 allele and lower in those carrying an epsilon 2 allele, but the effect on cerebrovascular disease is controversial. We estimated the risk of stroke associated with different apoE genotypes in older persons. METHODS: At the sixth annual follow-up of the Iowa cohort of the Established Populations for Epidemiologic Studies of the Elderly, 1664 persons aged > or = 71 years and free of stroke were genotyped for apo E. Occurrence of ischemic strokes was prospectively assessed from subsequent hospital discharge records and death certificates. RESULTS: One hundred fifty persons had an ischemic stroke over the subsequent 5 years (21.2 per 1000 person-years). The presence of epsilon 3 and epsilon 4 did not influence stroke risk. Among persons aged < 80 years at the time of genotyping, epsilon 2 carriers had lower risk of incident stroke, while no effect was detected in the older group. Compared with epsilon 2 carriers aged 70 to 79 years (reference group), those in the same age group and not carrying an epsilon 2 had 2.6-fold higher risk of incident stroke, and those aged > or = 80 years had even higher risk of stroke but without any difference according to presence/absence of epsilon 2 (relative risks 3.6 and 3.3). Results remained substantially unchanged when adjusted for potential confounders and in models estimating the effect of apoE polymorphism on the risk of developing a stroke at ages between 70 and 79 years (56 events) and separately at ages > or = 80 years (94 events). CONCLUSIONS: The conditioning influence of age on the protection conferred by the apoE epsilon 2 allele on stroke risk may account for previous controversies. This hypothesis should be verified in a population with a wider age range. PMID- 9412624 TI - A mutation in plasma platelet-activating factor acetylhydrolase (Val279-->Phe) is a genetic risk factor for stroke. AB - BACKGROUND AND PURPOSE: Platelet-activating factor (PAF) is a phospholipid with multiple actions that include thrombosis and inflammation. It is inactivated by a plasma enzyme, PAF acetylhydrolase. Deficiency of this enzyme in plasma is caused by a missense mutation in the gene (Val279-->Phe). We have studied a possible association of this mutation with the risk of stroke. SUBJECTS AND METHODS: We studied 120 consecutive patients with cerebral thrombosis. The control group consisted of 134 patients matched for age and sex with minor complaints but without stroke. Genomic DNA was analyzed for the mutant allele by a specific polymerase-chain reaction. Plasma PAF acetylhydrolase activity was determined by the method of Stafforini et al. RESULTS: The prevalence of the mutant gene was 43.4% in stroke patients (39.2% heterozygotes and 4.2% homozygotes), which was significantly higher than the 25.4% in control subjects (22.4% heterozygotes and 3.0% homozygotes) (chi 2 = 9.22, P < .01). The prevalence was slightly higher in stroke patients without hypertension than those with hypertension, but the difference was not significant. The patients with family histories of stroke had a slightly higher but not a significant prevalence of the mutant gene as compared with those without family histories of stroke. Plasma PAF acetylhydrolase activity was higher in patients than in control subjects, in normal subjects, or patients with a heterozygous genotype. CONCLUSIONS: These results suggest that plasma PAF acetylhydrolase deficiency may be a risk factor for stroke. This may explain the relatively high prevalence of stroke in Japan, as the mutation is more common among Japanese than Caucasians. PMID- 9412625 TI - Pseudoxanthoma elasticum lesions and cardiac complications as contributing factors for strokes in beta-thalassemia patients. AB - BACKGROUND AND PURPOSE: Pseudoxanthoma elasticum (PXE) lesions, which lead to intracranial hemorrhages and cardiac complications, predisposing to thrombotic strokes, are frequent findings in beta-thalassemia. Nevertheless, the association of these lesions with strokes in thalassemic patients has not been previously discussed. METHODS: Ten beta-thalassemic patients who developed an intracranial hemorrhage or a thrombotic stroke were reviewed. RESULTS: In the group of the four patients presenting with hemorrhage, one had PXE lesions, one had cardiac abnormalities, and one both PXE and cardiac disorders. In the group presenting with thrombotic stroke, all six patients had cardiac abnormalities and platelet count elevation due to splenectomy. Three also had PXE findings. No other predisposing factor for stroke was present. CONCLUSIONS: Cardiac complications and PXE may be risk factors for strokes in beta-thalassemia. Their frequent coexistence leads to a therapeutic dilemma in patients requiring antithrombotic therapy. PMID- 9412626 TI - Superoxide dismutase activity in serum of patients with acute cerebral ischemic injury. Correlation with clinical course and infarct size. AB - BACKGROUND AND PURPOSE: Superoxide dismutase (SOD) is one of the major free radical scavenging systems that might play a role in both degenerative and acute diseases of the central nervous system. METHODS: We measured SOD activity in the serum of 41 patients with acute ischemic stroke with a chemiluminometric assay based on the generation of oxygen free radicals by xanthine and xanthine oxidase. RESULTS: SOD activity was significantly lower in patients with ischemic stroke than in age-matched control patients with nonvascular, neurological illnesses (n = 24; P < .034, Wilcoxon rank test). The activity was inversely correlated with the size of infarction on CT (P = .01, Spearman correlation) and the severity of neurological deficits (P < .001, Spearman correlation). The decreased SOD activity recovered within 5 days after stroke to values found in serum of control patients. CONCLUSIONS: Our data suggest that the SOD activity in serum is reduced in stroke patients, and replacement of antioxidative activity could be beneficial in the acute treatment of cerebral ischemia. PMID- 9412627 TI - Preexisting dementia in stroke patients. Baseline frequency, associated factors, and outcome. AB - BACKGROUND AND PURPOSE: The link between stroke and degenerative dementia, especially Alzheimer's disease, is closer than expected by chance. Dementia after stroke may be due to the cumulative effect of vascular and degenerative changes. The prevalence of dementia just before stroke onset remains unsettled. The aim of this study was to determine the frequency of preexisting dementia in stroke patients, associated factors, and consequences on outcome. METHODS: We evaluated the cognitive functioning prior to stroke in 202 consecutive patients with ischemic or hemorrhagic stroke by means of the Informant Questionnaire on Cognitive Decline in the Elderly (IQCODE). We classified in the dementia group patients with IQCODE scores of 104 or more. Six months after stroke onset, survivors underwent a battery of neuropsychological tests. RESULTS: Thirty-three patients were demented before stroke (16.3%; 95% confidence interval, 11.2 to 21.4). There was no diagnosis of dementia in 32 of these 33 patients. We determined by logistic regression analysis that female sex, family dementia, leukoaraiosis, and cerebral atrophy are independently associated with prestroke dementia. All survivors who had IQCODE scores of 104 or more at the acute stage met criteria for dementia 6 months later. CONCLUSIONS: Our study showed that one sixth of stroke patients have preexisting dementia. Therefore, some patients with so-called "poststroke dementia" probably had unrecognized preexisting dementia. PMID- 9412628 TI - Poststroke apathy and regional cerebral blood flow. AB - BACKGROUND AND PURPOSE: Although apathy has been reported as one of the neuropsychiatric symptoms following stroke, there have been no studies on regional cerebral blood flow (rCBF) in patients with apathy. In this study we estimated the severity of apathy using the Apathy Scale and examined its relationship to rCBF in 40 stroke patients (mean age, 71.4 years). METHODS: Neuropsychiatric batteries were performed including the Apathy Scale, verbal intelligence and frontal function tests, a depression scale, and an assessment of activities of daily living. The cortical rCBFs were measured by the 133Xe inhalation method. RESULTS: Twenty patients (50%) showed apathy. These patients showed significantly lower scores on verbal intelligence and frontal function tests and a significantly higher depression score than the nonapathetic group. On MRI images there was no relationship between the apathy score and specific regional distribution of lesions. The rCBFs of the bilateral hemisphere were significantly lower in the apathetic group than in the nonapathetic group. The apathetic group showed a significantly reduced rCBF in the right dorsolateral frontal and left fronto-temporal regions. Furthermore, the apathy score for all patients was significantly negatively correlated with rCBF in the same regions. CONCLUSIONS: These findings demonstrate that apathy is a frequent symptom among elderly stroke patients and may be accompanied by cognitive impairments, depressive state, and frontal dysfunction. The hypoactivity in the frontal lobe and anterior temporal regions may contribute to symptoms of apathy after stroke. PMID- 9412629 TI - Increased common carotid intima-media thickness. Adaptive response or a reflection of atherosclerosis? Findings from the Rotterdam Study. AB - BACKGROUND AND PURPOSE: Carotid intima-media thickness (IMT) measurements are used widely to study atherosclerosis. Some have suggested that an increased IMT reflects a nonatherosclerotic adaptive response to changes in shear stress and tensile stress. This stems from the hypothesis that changes in shear stress and subsequently in lumen diameter are followed by changes in IMT to keep tensile stress constant. We studied the relation of common carotid IMT to common carotid end-diastolic lumen diameter and tensile stress, as approximated by mean arterial pressure (lumen diameter/IMT)]. METHODS: A cross-sectional analysis was performed with data obtained from the first 1715 participants in the Rotterdam Study, a population-based cohort study among 7983 subjects aged 55 years and over who underwent ultrasonographic examination of the carotid arteries. End-diastolic lumen diameter and IMT of the common carotid arteries were evaluated and quantified. RESULTS: With increasing IMT, inner and outer lumen diameters increased gradually, and beyond an IMT of 1.10 mm, the inner lumen diameter decreased. Tensile stress increased with increasing lumen diameter instead of being constant. The lumen-to-IMT ratio was constant across levels of mean arterial pressure. CONCLUSIONS: Our findings are compatible with the view that at lower degrees of IMT, the thickening appears to reflect an equilibrium state in which the effects of pressure and flow on the arteries are in balance, given a characteristic relation between shear stress and local transmural pressure. Beyond a certain level, IMT more likely may represent atherosclerosis. Regardless of whether common carotid IMT reflects local atherosclerosis, it may still serve as a graded marker for cardiovascular risk. PMID- 9412630 TI - Role of transcranial Doppler and stump pressure during carotid endarterectomy. AB - BACKGROUND AND PURPOSE: The aim of our study was to clarify the pathophysiology of perioperative cerebral complications during carotid endarterectomy in our series. METHODS: By means of transcranial Doppler ultrasonography and stump pressure measurement, we monitored 112 patients who underwent carotid endarterectomy under general anesthesia for symptomatic or asymptomatic severe carotid stenosis. RESULTS: Of 18 patients who underwent carotid endarterectomy with intra-arterial shunt, 2 (11.1%) developed an ischemic stroke. Of the other 94 patients, one suffered a nucleocapsular hemorrhage and 5 had cerebral ischemic complications. In these 5 patients, the duration of clamping was significantly longer (mean +/- SD, 16.4 +/- 1.1 versus 12.7 +/- 2.6 minutes; P = .0019), and the decrease of middle cerebral artery mean velocity on clamping was significantly greater (mean +/- SD, 56.4 +/- 4.9% versus 28.8 +/- 20.2%; P = .0031), while stump pressure was not significantly different. Microembolic signals were recorded in 70 patients (62.5%) and were not associated with cerebral ischemic complications. The 7 patients who developed cerebral ischemic complications had a significantly higher percentage of stenosis in the contralateral internal carotid artery (mean +/- SD, 82.0 +/- 17.8% versus 29.3 +/ 36.4%; P = .0018). CONCLUSIONS: The results of our study suggest that the major complications of carotid endarterectomy may be due to hemodynamic factors. Stump pressure alone is not a reliable indicator of hemodynamic changes that predict cerebral ischemia. Particulate microembolism may cause more subtle changes in cerebral parenchyma, but further studies are needed to clarify this point. PMID- 9412631 TI - Oxygen inhalation can differentiate gaseous from nongaseous microemboli detected by transcranial Doppler ultrasound. AB - BACKGROUND AND PURPOSE: Clinically silent circulating microemboli can be detected by transcranial Doppler sonography. The composition of these emboli in different clinical conditions is unclear. METHODS: We performed 1-hour transcranial Doppler sonographic recordings from the middle cerebral or posterior cerebral artery in 20 patients with mechanical prosthetic heart valves, in 78 patients with an arterial embolic source, and in 20 control subjects. During 30 minutes of this recording, the patients inspired room air and 6 L of oxygen per minute via a loosely fitting facial mask; during the remaining 30 minutes, they breathed room air only. RESULTS: There was a significant decline of embolic signals (ES) under oxygen in the patients with mechanical prosthetic cardiac valves (144 ES without oxygen versus 63 ES with oxygen; P = .002) but not in the patients with arterial embolic sources (145 ES without oxygen versus 135 ES with oxygen; P = NS). In the control subjects, no ES were found. CONCLUSIONS: ES in patients with mechanical prosthetic cardiac valves correspond mainly to gas bubbles. Oxygen inhibits the cavitation process of mechanical prosthetic heart valves or speeds up redissolution of gas bubbles generated by cavitation. In contrast, solid microemboli originating from thrombus or atheroma cannot be suppressed by oxygen inhalation. This simple method of oxygen inhalation should help to clarify the composition of microemboli in various clinical and experimental settings. PMID- 9412632 TI - Spontaneous oscillations in cerebral blood flow velocities in middle cerebral arteries in control subjects and patients with epilepsy. AB - BACKGROUND AND PURPOSE: Cardiac arrhythmias mediated by the sympathetic nervous system have been implicated in sudden, unexplained deaths in patients with epilepsy. Cerebral blood flow velocities (CBFV) as measured by transcranial Doppler are characterized by slow spontaneous oscillations in part attributed to changes in sympathetic activity (M waves, 3 to 9 cycles per minute) and to discharges of monoaminergic neurons in the brain stem (B waves, 0.5 to 2 cycles per minute). This study was designed to compare spontaneous fluctuations of CBFV in patients with epilepsy with those in normal control subjects. METHODS: Simultaneous registrations of scalp electroencephalograms, with electrodes placed according to the 10-20 System, and transcranial Doppler recordings of both middle cerebral arteries were performed in 27 patients (9 with primary generalized epilepsy, 18 with focal epilepsy). Data analysis of CBFV was based on the envelope curves of the Doppler spectrum. A fast Fourier transformation over the 20-minute CBFV curve was performed, and the amplitudes of B and M waves were calculated and compared with those in 20 normal, age-matched control subjects. RESULTS: While the amplitudes of the B waves in both groups were similar, patients with epilepsy showed significantly increased M waves. Patients with focal epilepsy did not present asymmetries between the normal hemisphere and the side of the epileptic focus with respect to both M and B waves. CONCLUSIONS: Enhanced M waves in epileptic patients may reflect increased sympathetic activity even in the absence of seizures. This study provides further evidence for an autonomic dysfunction as a possible mechanism for sudden unexplained death in patients with epilepsy. PMID- 9412633 TI - Comparison of hemodynamic cerebral ischemia and microembolic signals detected during carotid endarterectomy and carotid angioplasty. AB - BACKGROUND AND PURPOSE: There has been concern about carotid percutaneous transluminal angioplasty (PTA) carrying a greater risk of cerebral ischemia than carotid endarterectomy. We set out to compare cerebral hemodynamics and microembolization during carotid PTA and CEA. METHODS: We used transcranial Doppler to monitor the middle cerebral artery of 28 patients undergoing carotid PTA (n = 14) or carotid endarterectomy (CEA) with a shunt (n = 14). Each period during which the internal carotid artery was occluded by PTA balloon or by clamp when the shunt was not in place was timed. Individual periods were summated to give a total occlusion time. Ischemic time was defined as the period for which mean middle cerebral artery velocity fell to a third or less of baseline. Microembolic signals were counted during each procedure. RESULTS: CEA resulted in significantly longer individual and total occlusion time than PTA (mean individual occlusion time, seconds), CEA, 168 +/- 51; PTA, 20 +/- 7; P < .001; mean total occlusion time; CEA, 337 +/- 70; PTA, 26 +/- 10; P < .001. Ischemic time was also significantly longer during CEA than during PTA (CEA, 165 +/- 40; PTA, 17 +/- 5; P = .001). There were significantly more microembolic signals during PTA than during CEA (mean number of microembolic signals during CEA, 52 +/ 64; during PTA, 202 +/- 119; P = .001). There was no correlation between any of the parameters measured and periprocedural stroke, which occurred in one patient in each group. CONCLUSION: PTA results in less hemodynamic ischemia but more cerebral microembolism than CEA. In this small series, however, it is not possible to comment on the relations between ischemic time, microembolism, and stroke. PMID- 9412634 TI - Noninvasive prediction of intracranial pressure curves using transcranial Doppler ultrasonography and blood pressure curves. AB - BACKGROUND AND PURPOSE: Until now the assessment of intracranial pressure (ICP) required invasive methods. The objective of this study was to introduce an approach to a noninvasive assessment of continuous ICP curves. METHODS: The intracranial compartment was considered a "black box" system with an input signal, the arterial blood pressure (ABP), and an output signal, the ICP. A so called weight function described the relationship between ABP and ICP curves. Certain parameters, called transcranial Doppler (TCD) characteristics, were calculated from the cerebral blood flow velocity (FV) and the ABP curves and were used to estimate this weight function. From simultaneously sampled FV, ABP, and (invasively measured) ICP curves of a defined group of patients with severe head injuries, the TCD characteristics and the weight function were computed. Multiple regression analysis revealed a mathematical formula for calculating the weight function from TCD characteristics. This formula was used to generate the ICP simulation. FV, ABP, and ICP recordings from 11 patients (mean age, 46 +/- 14 years) with severe head injury were studied. In each patient, ICP was computed by a simulation procedure, generated from the data of the remaining 10 patients. The simulation period was 100 seconds. RESULTS: Corresponding pressure trends with a mean absolute difference of 4.0 +/- 1.8 mm Hg between computed and measured ICP were observed. Shapes of pulse and respiratory ICP modulations were clearly predicted. CONCLUSIONS: These results demonstrate that this method constitutes a promising step toward a noninvasive ICP prediction that may be clinically applicable under well-defined conditions. PMID- 9412635 TI - Contrast-enhanced transcranial color-coded duplex sonography in ischemic cerebrovascular disease. AB - BACKGROUND AND PURPOSE: Echo contrast agents have been shown to provide conclusive examinations in most patients with insufficient ultrasound penetration through the temporal bone. We investigated the diagnostic value of contrast enhanced (CE) transcranial color-coded duplex sonography (TCCD) in patients with ischemic cerebrovascular disease and insufficient temporal windows and evaluated TCCD criteria that predict whether CE-TCCD studies may become conclusive. METHODS: Thirty-three patients presenting with ischemic strokes (n = 21) and transient ischemic attacks (n = 12) were investigated. Extracranial color duplex imaging showed normal findings in 24 patients, eight > or = 70% stenoses and one occlusion of the carotid arteries in 8 patients, and severe occlusive disease of both vertebral arteries in 1 patient. Seven carotid stenoses and vertebral artery obstructions were confirmed by angiography. The galactose/palmitic acid-based echo contrast agent was injected intravenously as bolus of 200, 300, or 400 mg/mL in a dosage of 10, 5, and 5 mL, respectively. RESULTS: Thirty-two of the 33 patients were completely examined because 1 patient who felt pain at the injection site declined further investigations. Twenty-one (66%) of 32 CE studies were conclusive and showed cross-flow through three anterior and two posterior communicating arteries, but no stenoses and occlusions. Precontrast identification of any cerebral artery provided an overall accuracy of 97% in predicting a conclusive CE investigation. Precontrast TCCD identified no arterial Doppler signals in patients with inconclusive CE studies. CONCLUSIONS: CE-TCCD provided conclusive examinations in two thirds of patients with ischemic cerebrovascular disease and ultrasound-refractory temporal windows. Precontrast detection of any cerebral artery reliably predicted a conclusive CE investigation. PMID- 9412636 TI - Cerebrovascular reserve capacity many years after vasospasm due to aneurysmal subarachnoid hemorrhage. A transcranial Doppler study with acetazolamide test. AB - BACKGROUND AND PURPOSE: Vasospasm in aneurysmal subarachnoid hemorrhage results in proliferative vasculopathy. Systemic hypertension also causes vascular hypertrophy. Both of these histological changes can lead to rigidity of the cerebrovascular system, reducing its autoregulatory capacity. METHODS: Blood flow velocity (BFV) in the middle cerebral artery at rest and cerebrovascular reserve capacity (CVRC) (percent rise in BFV after acetazolamide stimulation) measured by means of transcranial Doppler sonography were studied many years after aneurysmal subarachnoid hemorrhage in patients with proven cerebral vasospasm (mean BFV > 160 cm/s). The BFV under resting conditions and the CVRC values of the ipsilateral and the contralateral hemispheres were measured in 29 patients (mean age, 43 years; mean follow-up, 4.6 years) and compared with those of control subjects. RESULTS: Persistent high BFV (> 120 cm/s) was found in three patients in the peripheral branch of the ipsilateral middle cerebral artery. In the main trunks of the arteries of the anterior circle of Willis, BFV was normal in all cases. CVRC was normal in all patients (ipsilateral, 52 +/- 21%; contralateral, 56 +/- 17%); values did not differ significantly from each other or from the control value (45 +/- 18%). The higher value of CVRC on the contralateral side was found to be statistically significant in selected groups (hypertensive patients and patients with residual infarct on late CT). CONCLUSIONS: Proliferative vasculopathy developed at the time of vasospasm must have resolved and did not reduce late vasoreactivity. Comorbidity with hypertension also did not seem to influence the late vasoreactivity toward normalization. PMID- 9412637 TI - Interobserver agreement for 10% categories of angiographic carotid stenosis. AB - BACKGROUND AND PURPOSE: Although the reliability of the assessment of severe 70% to 99% carotid stenosis by carotid angiography has been proven excellent, this may not necessarily be the case for a more detailed classification of carotid stenoses by 10% categories. METHODS: Angiograms of the carotid arteries were assessed pairwise by three independent, experienced observers. The measurements of the degree of stenosis of both the carotid bifurcation and the internal carotid artery were made according to the European Carotid Surgery Trial method. Kappa statistics were used to assess the agreement beyond chance for severe (70% to 99%) carotid stenosis (kappa 1) and for 10% categories of carotid stenosis (kappa 2). The penalty scores were adjusted by weights for the relative difference in risk (RDR) of stroke in the ipsilateral carotid distribution between the 10% categories (kappa 3). An adjustment of the RDR method was made by assuming that only patients with a severe carotid stenosis would undergo surgery, and the penalty would be 0 if no disagreement would exist about the indication for surgery (kappa 4). An even further adjustment (kappa 5) was made by assuming that assessment of the rate of carotid stenosis by one or both observers would lead to different treatment recommendations in 50% of the cases, and accordingly the penalty for disagreement (RDR) was halved. RESULTS: One hundred twenty-one carotid bifurcations in 65 patients with a transient ischemic attack or nondisabling stroke were assessed. The intraclass correlation between the exact estimates of carotid stenosis was .90 (95% confidence interval, .85 to .92). The mean difference in stenosis between the two raters was 0.8% (95% confidence interval, -2.1% to 3.7%). kappa 1 to kappa 5 equaled 0.80, 0.40, 0.79, 0.91, and 0.92, respectively. CONCLUSIONS: Interobserver agreement for distinct 10% categories of angiographic carotid stenosis is moderate, but when realistic risk- and decision-based weights are used, agreement between experienced observers can be almost perfect. PMID- 9412638 TI - Diameter dependence of myogenic tone of human pial arteries. Possible relation to distensibility. AB - BACKGROUND AND PURPOSE: Responses to changes in intraluminal pressure of isolated human pial arteries (200 to 1200 microns i.d.) obtained from patients undergoing neurosurgery were measured. METHODS: The vessels were cannulated and pressurized (60 mm Hg); vascular diameter and intraluminal pressure were recorded simultaneously. After spontaneous development of steady state tone, intraluminal pressure was changed to both higher and lower levels in random sequence. RESULTS: Human pial arteries exhibited myogenic responses and maintained their diameter over the pressure range of 20 to 100 mm Hg. The level of myogenic tone observed at 30 mm Hg did not vary significantly with artery diameter. In contrast, at 60 and 90 mm Hg, the extent of myogenic tone increased as the diameter decreased (up to 70% to 80% of maximum in 200-microns i.d. arteries). The arteries contracted to KCl 30 mmol/L, norepinephrine 1 mumol/L, and vasopressin 0.1 mumol/L and relaxed to acetylcholine 3 mumol/L. The extent of these responses did not vary with the diameter of the artery. Arterial distensibility, represented by the slope of the tangent of the passive pressure-diameter curve at lower pressures (5 to 50 mm Hg), increased as arteries became smaller. This is consistent with the possibility that the level of myogenic tone is related to vessel distensibility. Human omental arteries of comparable size did not develop myogenic tone but contracted to KCl and norepinephrine and relaxed to acetylcholine to an extent similar to pial arteries. CONCLUSIONS: There is a specific gradient of myogenic responsiveness in human pial arteries that varies inversely with their diameter. This tone does not develop in all vascular beds. These levels of tone in the pial circulation would be expected to be of profound functional significance by allowing blood flow to vary widely. PMID- 9412639 TI - Trends in case-fatality of stroke in Finland during 1983 to 1992. AB - BACKGROUND AND PURPOSE: Stroke mortality has been declining in Finland during the past 20 years. It is not known, however, whether this favorable development is attributable to the decline in the incidence or case-fatality of stroke. For this reason we examined the trends in case-fatality of stroke, including trends by subtype of stroke. METHODS: The analyses were carried out using data of the community-based FINMONICA Stroke Register, which was operating in three geographic areas of Finland during 1983 to 1992. All stroke events (n = 11,171) in persons aged 35 to 74 years were included in this register. RESULTS: The 28 day case-fatality of stroke fell yearly by 3.6% (P = .01) in men and by 2.6% (P = .2) in women. At the end of the study period, the average 28-day case-fatality of all strokes was 20% in men and 21% in women. Considerable differences by subtype of stroke were observed. The 28-day case-fatalities at the end of the study period were in men-56% for subarachnoid hemorrhage, 42% for intracerebral hemorrhage, and 14% for cerebral infarction. In women, the corresponding figures were 49%, 49%, and 14%. The 28-day case-fatality of subarachnoid hemorrhage did not change during the study period, but for intracerebral hemorrhage, a significant decline was observed in men and there was a declining trend also in women. The 28-day case-fatality of cerebral infarction declined significantly in both genders. CONCLUSIONS: With the exception of subarachnoid hemorrhage, the 28 day case-fatality of stroke has fallen in Finland. It is likely that this fall has contributed to the decline in stroke mortality. PMID- 9412640 TI - High stroke incidence in the prospective community-based L'Aquila registry (1994 1998). First year's results. AB - BACKGROUND AND PURPOSE: Changes in stroke incidence are likely to occur as a consequence of aging of the population, but evidence for this hypothesis is lacking. METHODS: A prospective community-based registry of first-ever strokes (1994 to 1998) classified according to the International Classification of Diseases, 9th Revision (ICD-9) was established in the L'Aquila district, central Italy, with a total population of 297,838 (1991 census). Patients were identified by active monitoring of multiple sources, including general practitioners. RESULTS: In 1994, 819 patients (398 men and 421 women; mean +/- SD age, 74.8 +/- 11.3 years) suffered from a first-ever stroke. Eighty-nine percent of the patients had neuroimaging studies of the brain and were reclassified with the recent Application of the International Classification of Diseases to Neurology (ICD-10 NA). The occurrence of subarachnoid hemorrhage, intracerebral hemorrhage, cerebral infarction, and ill-defined events was 2.9%, 14.9%, 80.2%, and 2.0%, respectively. Crude annual incidence of first-ever stroke was 2.75/1000 (95% confidence interval [CI], 2.57 to 2.94) and 24.23/1000 (95% CI, 21.65 to 27.10) in patients older than 80 years. Incidence rates were higher in men and steeply increased with age. The standardized rate was 2.37/1000 for the Italian and 2.28/1000 for the European population. The 30-day case-fatality rate was 25.6% (95% CI, 22.8% to 28.7%). The occurrence of death, disability, and full recovery at 1 year was 36.9%, 38.9%, and 24.2%, respectively. No differences were found in stroke incidence and case-fatality according to income and urban or rural residences. CONCLUSIONS: In our population-based study, we found a high stroke incidence notably in the older age subgroups, suggesting that rather than declining, stroke is only being postponed until later in life. PMID- 9412641 TI - Subtypes of ischemic stroke. A hospital-based stroke registry in Taiwan (SCAN IV). AB - BACKGROUND AND PURPOSE: To better understand the clinical pattern and further elucidate the risk factors and outcome in different subtypes of cerebral infarction (CI) of the Chinese in Taiwan, we analyzed the National Taiwan University Hospital Stroke Registry in 1995 and performed an ethnic comparison with similar data banks. METHODS: From the National Taiwan University Hospital Stroke Registry in 1995, 676 patients (383 men and 293 women; mean age, 64.9 years; SD, 13.8 years; range, 1 to 98 years) with CI were recruited for this analysis. CI was classified into five subtypes based on clinical manifestations, ultrasonographic studies, and neuroimaging findings: large-artery atherosclerosis, lacunae, cardioembolism, other less common determined causes, and undetermined cause. Vascular risk factors, extracranial carotid artery atherosclerosis, and 30-day case-fatality rates were investigated in each subtype of CI. RESULTS: Of all CI patients, 17%, 29%, 20%, 6%, and 29% were classified as large-artery atherosclerosis, lacunae, cardioembolism, other determined causes, and undetermined cause subtypes, respectively. The present results were compared with those from eight similar Western stroke registries. The relative incidence of lacunar CI in Chinese patients was more common, but large-artery atherosclerotic CI was less common than in whites. Hypertension was frequently seen in CI patients, especially in those with lacunae (85%) and large-artery atherosclerosis (69%). Patients with cardioembolism had a higher percentage of atrial fibrillation (69%), left ventricular hypertrophy, and ischemic heart disease than the other patients. Patients with large-artery atherosclerosis had more vascular risk factors, such as hypertension, diabetes mellitus, smoking, and carotid stenosis. Cardioembolic patients had higher case-fatality rates than other CI patients. Of the cardioembolic patients, 17.3% and 21.8% died within 30 days and during hospitalization, respectively. CONCLUSIONS: The proportion of CI subtypes varied in different stroke registries. This may be partly due to applied classification criteria and racial-ethnic differences. Awareness of the risk factors and outcome in each subtype of stroke may afford further insights into the surveillance and treatment of cerebrovascular disease. PMID- 9412642 TI - A clinical comparison of definite moyamoya disease between South Korea and Japan. AB - BACKGROUND AND PURPOSE: The goal of the present study was to clarify whether South Korean patients with moyamoya disease have clinical features similar to those of Japanese patients. METHODS: From 26 South Korean neurosurgical institutes, 296 definite cases were collected and analyzed statistically. These cases were then compared with 731 Japanese definite cases registered to the Research Committee on Moyamoya Disease of the Ministry of Health and Welfare, Japan. RESULTS: The Korean age distribution patterns showed two peaks that were similar to those seen in Japanese patients. The incidence of adult moyamoya disease in South Korea, however, was 20% higher than that in Japanese patients. The family occurrence rate was 1.8% in Koreans. The incidence of cerebral infarction and bleeding in Koreans was statistically higher, whereas transient ischemic attack and seizure were less than those in Japanese subjects. The incidence of infarction in children and that of hemorrhage in both children and adults were also statistically higher in Koreans. The incidence of hemorrhage was higher in females than in males. Both the age at onset and sex affected the disease type. Although most Japanese patients underwent direct bypass surgery and/or combined indirect bypass procedures, single encephaloduroar teriosynangiosis was performed on 87.6% of all surgical cases in Koreans. Despite the higher incidence of hemorrhagic type in South Korea, the outcomes of the patients were similar to those of the Japanese patients. CONCLUSIONS: This study suggests that the clinical background of moyamoya disease in South Korea is essentially similar to that in Japan. PMID- 9412643 TI - A functional MRI study of subjects recovered from hemiparetic stroke. AB - BACKGROUND AND PURPOSE: Stroke recovery mechanisms remain incompletely understood, particularly for subjects with cortical stroke, in whom limited data are available. We used functional magnetic resonance imaging to compare brain activations in normal controls and subjects who recovered from hemiparetic stroke. METHODS: Functional magnetic resonance imaging was performed in ten stroke subjects with good recovery, five with deep, and five with cortical infarcts. Brain activation was achieved by index finger-tapping. Statistical parametric activation maps were obtained using a t test and a threshold of P < .001. In five bilateral motor regions, the volume of activated brain for each stroke subject was compared with the distribution of activation volumes among nine controls. RESULTS: Control subjects activated several motor regions. During recovered hand finger-tapping, stroke subjects activated the same regions as controls, often in a larger brain volume. In the unaffected hemisphere, sensorimotor cortex activation was increased in six of nine stroke subjects compared with controls. Cerebellar hemisphere contralateral and premotor cortex ipsilateral to this region, as well as supplementary motor areas, also had increased activation. In the stroke hemisphere, activation exceeding controls was uncommon, except that three of five cortical strokes showed peri-infarct activation foci. During unaffected hand finger-tapping, increased activation by stroke subjects compared with controls was uncommon; however, decreased activation was seen in unaffected sensorimotor cortex, suggesting that this region's responsiveness increased to the ipsilateral hand and decreased to contralateral hand movements. Use of a different threshold for defining activation (P < .01) did not change the overall findings (kappa = .75). CONCLUSIONS: Recovered finger-tapping by stroke subjects activated the same motor regions as controls but to a larger extent, particularly in the unaffected hemisphere. Increased reliance on these motor areas may represent an important component of motor recovery. Functional magnetic resonance imaging studies of subjects who recovered from stroke provide evidence for several processes that may be related to restoration of neurologic function. PMID- 9412644 TI - Cerebral magnetic resonance T2 high intensities in end-stage renal disease. AB - BACKGROUND AND PURPOSE: The pathophysiological mechanisms that cause cerebral MR T2 high intensities in end-stage renal disease (ESRD) are unclear. We evaluated the incidence and the risk factors of T2-weighted MR brain high intensities in patients with ESRD. METHODS: We examined the degree of T2-weighted MR brain high intensities (high intensity score) and determined the variables that had an independent association with the occurrence of high intensities in 38 patients with ESRD before chronic dialysis treatment, 173 patients with essential hypertension, and 72 normotensive control subjects. RESULTS: The whole brain high intensity score was significantly higher in patients with ESRD than in the control subjects, but there was no significant difference in high intensity score between the ESRD and the hypertensive groups. Age, hypertension, and smoking were significant independent predictors of high intensities in a multiple logistic regression model. The distribution pattern of high intensities in ESRD patients was very similar to that obtained from hypertensive patients; the high intensity score was highest in the corona radiata and was lowest in the cerebellum. CONCLUSIONS: T2 high intensities on MR images of ESRD may reflect subcortical small-vessel alterations induced by hypertension. PMID- 9412645 TI - Spinal cord ischemic injury. Development of a new model in the rat. AB - BACKGROUND AND PURPOSE: Spinal cord ischemic injury (SCII) with resulting paralysis is a major cause of morbidity after operations on the thoracic aorta. Since the vascular supply to the spinal cord is similar in rats and humans, the rat appears important for studies of mechanisms of injury and development of therapeutic strategies to avoid this complication. METHODS: In group A rats, we induced SCII using a previously described method, by occluding the descending thoracic aorta for 15, 20, 24, or 30 minutes with the inflated balloon of a 2F Fogarty catheter inserted through the femoral artery. In group B, the catheter was inserted through the left common carotid artery, and the aorta was occluded just distal to the carotid origin for 20 minutes. In group C, in addition to the procedure described for group B, hypovolemia was induced during a 12-minute period of aortic occlusion by equilibrating the left femoral artery pressure to the atmospheric pressure. The motor function of the hind limbs and the associated spinal cord histopathology were studied. RESULTS: At 96 hours, 9 of 10 rats in group C were paraplegic. This rate was significantly higher than that of group A (1 of 21, P = .00000) or group B (4 of 10, P < .03). In all groups, the histopathological changes became more severe from the rostral to the caudal direction along the spinal cord and from the peripheral to the central location in transverse sections. CONCLUSIONS: The combination of aortic arch occlusion with induced hypovolemia resulted in a reproducible model of SCII in rats. PMID- 9412646 TI - The calmodulin antagonist trifluoperazine in transient focal brain ischemia in rats. Anti-ischemic effect and therapeutic window. AB - BACKGROUND AND PURPOSE: This study was performed to assess the efficacy and the therapeutic window for the calmodulin antagonist trifluoperazine in experiments involving transient middle cerebral artery (MCA) occlusion. METHODS: Male Wistar rats were subjected to transient (2 hours) MCA occlusion by an intraluminal filament technique. Trifluoperazine (5.0 mg.kg-1) was injected intraperitoneally 5 minutes, 1 hour, or 2 hours after the induction of ischemia. Drug administration was repeated 24 hours after the first injection. Neurological scores and infarct volumes were evaluated at 48 hours of reperfusion. The effect of trifluoperazine on cortical blood flow was studied with continuous laser Doppler flowmetry. RESULTS: The median value of neurological scores in the control rats (n = 7) was 3, while those in the treated groups were 1 (5-minute group; n = 7, P < .05) and 2 (1-hour and 2-hour groups; each n = 7). The percentage of infarct volume in the control rats was 34.8 +/- 4.9% (mean +/- SD), while those in the treated groups were 11.3 +/- 12.3% (5-minute group; P < .01), 24.8 +/- 15.1% (1-hour group), and 28.8 +/- 8.3% (2-hour group). Trifluoperazine, given at 5 minutes after ischemia, had no influence on blood flow in the neocortical penumbra during and after ischemia. CONCLUSIONS: The results demonstrate that trifluoperazine markedly reduces infarct volume after 2 hours of MCA occlusion when given 5 minutes after the induction of ischemia. However, the therapeutic window for trifluoperazine seems narrow since the drug had no significant effect when given after 1 or 2 hours. PMID- 9412647 TI - Differential hydroxylation of salicylate in core and penumbra regions during focal reversible cerebral ischemia. AB - BACKGROUND AND PURPOSE: Free radical-mediated damage during and/or after cerebral ischemia is thought to participate in the elaboration of stroke-related injury. To elucidate the role of this mechanism in cerebral damage, the study presented herein sought to clarify the spatial and temporal features of the free radical response to transient ischemia. With use of a reproducible model of in vivo focal ischemia/reperfusion, the time course of salicylate hydroxylation was measured in ischemic core and penumbra regions. METHODS: Transient focal cerebral ischemia was produced in Sprague-Dawley rats by occluding both carotid arteries and one middle cerebral artery for 3 hours, followed by reperfusion. Cerebral reperfusion was confirmed by visual inspection and iodo[14C]antipyrine autoradiography. A microdialysis probe was placed stereotactically in either the ischemic core or ischemic penumbra of the frontoparietal cortex; the probe was perfused with salicylate, and dialysate samples were analyzed by high-performance liquid chromatography for salicylate hydroxylation products. RESULTS: Salicylate hydroxylation was significantly increased during ischemia and was further increased during 6 hours of reperfusion in the penumbra compared with sham controls. In comparison, a delayed increase in hydroxylation was observed within the ischemic core region only after 3 hours of reperfusion. CONCLUSION: A differential generation of salicylate hydroxylation occurs in core and penumbra regions in association with focal ischemia/reperfusion of the rat neocortex. The early and progressive response in the penumbra suggests that free radical mechanisms may be continuously active in the aggravation of injury in the ischemic penumbra during ischemia and reperfusion. In contrast, the relatively delayed onset of hydroxylation in the core region indicates that this mechanism participates primarily in the late stages of ischemic injury in densely ischemic tissue. These findings are consistent with the concept that the role of free radicals in cerebral injury may differ qualitatively and/or quantitatively in areas of total and partial cerebral perfusion. PMID- 9412648 TI - Upper motor neuron lesions in stroke patients do not induce anterograde transneuronal degeneration in spinal anterior horn cells. AB - BACKGROUND AND PURPOSE: To determine whether upper motor neuron lesions in stroke can cause transneuronal degeneration of lower motor neurons, we assessed spinal anterior horn cells in patients dying with poststroke hemiplegia. METHODS: Subjects were four stroke patients with severe left hemiplegia and four age matched control subjects who died of nonneurological disease. After histological processing and staining, cytoarchitectonic assessment was made of all neurons in the ventral horns of the 4th lumbar segment of the spinal cord according to cell diameter and topography. RESULTS: In the four stroke patients, no differences were seen in anterior horn cell populations or diameter and size distribution patterns between affected and unaffected sides or between these patients and the control subjects. CONCLUSIONS: The present quantitative analysis provides no evidence of anterograde transneuronal degeneration of lower motor neurons after upper motor neuron damage in stroke patients. PMID- 9412649 TI - Effect of antihypertensive treatment in patients having already suffered from stroke. Gathering the evidence. The INDANA (INdividual Data ANalysis of Antihypertensive intervention trials) Project Collaborators. AB - BACKGROUND AND PURPOSE: Drug treatment of high blood pressure has been shown to reduce the associated cardiovascular risk. Stroke represents the type of event more strongly linked with high blood pressure, responsible for a high rate of death or invalidity, and with the highest proportion of events that can be avoided by treatment. Hypertensive patients with a history of cerebrovascular accident are at particularly high risk of recurrence. Specific trials of blood pressure lowering drugs in stroke survivors showed inconclusive results in the past. METHODS: We performed a meta-analysis using all available randomized controlled clinical trials assessing the effect of blood pressure lowering drugs on clinical outcomes (recurrence of stroke, coronary events, cause-specific, and overall mortality) in patients with prior stroke or transient ischemic attack. RESULTS: We identified 9 trials, including a total of 6752 patients: 2 trials included 551 hypertensive stroke survivors; 6 trials of hypertensive patients included a small proportion of stroke survivors (536 patients); 1 trial included stroke survivors, whether hypertensive or not (5665 patients). The recurrence of stroke, fatal and nonfatal, was significantly reduced in active groups compared with control groups consistently across the different sources of data (relative risk of 0.72, 95% confidence interval: 0.61 to 0.85). There was no evidence that this intervention induced serious adverse effect. CONCLUSIONS: Blood pressure lowering drug interventions reduced the risk of stroke recurrence in stroke survivors. Available data did not allow to verify whether such benefit depends on initial blood pressure level. More data are needed before considering antihypertensive therapy in normotensive patients at high cerebrovascular risk. PMID- 9412650 TI - Timing of carotid endarterectomy after stroke. AB - BACKGROUND: Timing of carotid endarterectomy after stroke in a patient with a fixed neurological deficit remains an important but unresolved question. Early surgery has been associated with cerebral hemorrhage and infarct extension. Delayed endarterectomy exposes the patient to recurrent stroke and carotid occlusion. This review investigates the hypothesis that timing of surgery after stroke influences outcome and complications. SUMMARY OF REVIEW: This analysis critically evaluates peer-reviewed reports that retrospectively examined outcome after surgery performed "early" and "late" after stroke. The basis for intracerebral hemorrhage after endarterectomy is discussed. Clinical features that influence outcome are investigated. CONCLUSIONS: Patients undergoing carotid endarterectomy are considered a heterogeneous group based on the following features: presence of low density on cranial CT, vascular territory of the infarct, brain shift, and level of consciousness. While critical review of these retrospective studies suggests that some patients with an acute stroke can safely undergo endarterectomy shortly after the diagnosis is made, direct answers to these questions of timing of endarterectomy after stroke are best addressed by prospective studies. Nevertheless, the present review provides a basis for decision making in certain patients and for the design of future investigations. PMID- 9412651 TI - Blood pressure profiles in acute stroke. PMID- 9412652 TI - Migraine: a risk factor for ischemic stroke in young women. PMID- 9412653 TI - Air emboli and prosthetic heart valves. PMID- 9412654 TI - Pretruncal subarachnoid hemorrhage: an anatomically correct description of the perimesencephalic subarachnoid hemorrhage. PMID- 9412655 TI - Lack of effectiveness of adjuvant alternating chemotherapy in node-positive, estrogen-receptor-negative premenopausal breast cancer patients: results of a multicentric Italian study. The Breast Cancer Adjuvant Chemo-Hormone Therapy Cooperative Group (GROCTA). AB - A multicentric randomized trial was performed in premenopausal women with node positive, estrogen-receptor-negative breast tumors to assess the potential superiority of alternating adjuvant chemotherapy over 'standard' CMF chemotherapy. Between January 1989 and June 1992, 107 patients were entered into the study and randomly allocated to receive either cyclophosphamide 100 mg/m2 per as on days 1-14, methotrexate 40 mg/m2 and 5-fluorouracil 600 mg/m2 intravenously (i.v.) on days 1, 8 (CMF), every 4 weeks for a total of 6 cycles, or the following regimens: CMF as previously; epidoxorubicin 75 mg/m2 i.v. on day 1 and vincristine 0.75 mg/m2 i.v. on days 1, 8 (EV); mitomycin-C 10 mg/m2 i.v. on day 1 and vindesine 2 mg/m2 i.v. on days 1, 8 (MVs). The three regimens were given every 4 weeks for a total of 6 cycles according to the following schedule: CMF, EV, MVs, CMF, EV, MVs. At a median follow up of 48 months (range 30-72), 40 patients have relapsed and 17 have died overall. More patients in the triple combination arm have relapsed and more have died, the latter difference tending toward statistical significance (p = 0.06). There was no statistical difference in the site of relapse between the two groups. Total duration of adjuvant therapy was similar in the two arms (312 chemotherapy cycles in the triple arm and 308 in the CMF arm). Treatment toxicity was also comparable, although more patients in the triple-combination arm were still regularly menstruating 6 months after the completion of chemotherapy. This study failed to show any advantage ensuing from the use of alternating chemotherapy in patients with early breast cancer. PMID- 9412656 TI - Expression of the HGF/SF receptor, c-met, and its ligand in human colorectal cancers. AB - The c-met proto-oncogene product is a receptor tyrosine kinase that mediates the effects of the multifunctional cytokine hepatocyte growth factor/scatter factor (HGF/SF). We have studied the expression of both the c-met receptor and HGF/SF at both the protein and message level in colorectal cancer tissues of varying disease stage. All of the tumors displayed an overexpression of the c-met mRNA compared to their normal tissue counterparts while 16 of 21 tissues (75%) displayed up-regulation of c-met protein. No HGF/SF mRNA or protein could be detected in either tissue type. Viable tumor cells extracted from cancer tissue exhibited increased motility in response to HGF/SF stimulation demonstrating that c-met was functionally active. No correlation between expression of c-met and tumor stage or degree of differentiation was observed. HGF/SF is known to be a potent stimulator of tumor cell motility and invasion, two cellular properties essential for the metastatic development of cancers. The overexpression of the HGF/SF receptor in colorectal cancers may result in an increased sensitivity to HGF/SF, which may confer an enhanced metastatic potential to the cancer cells within the tumor body. PMID- 9412657 TI - Antitumor effect of DT-5461, a lipid A derivative, against human tumor xenografts is mediated by intratumoral production of tumor necrosis factor and affected by host immunosuppressive factors in nude mice. AB - We previously reported that DT-5461, a synthetic low-toxic lipid A analog, inhibits growth of various murine tumors through activation of host immune systems. In the present study, DT-5461 also exhibited significant antitumor effects against 5 out of 6 human tumor xenografts in nude mice. The antitumor activity was similar to or greater than those of chemotherapeutics. Antitumor effects of DT-5461 significantly correlated with intratumoral levels of tumor necrosis factor (TNF) induced by the compound (r = 0.701, p < 0.05). In vitro TNF production by DT-5461-stimulated macrophages was augmented by tumor cells, and the augmentative effect correlated with TNF activity detected in these tumor tissues. Meanwhile, a weaker therapeutic efficacy of DT-5461 was observed against certain tumors that caused a significant increase in the level of immunosuppressive factors in host blood. These findings support the idea that intratumoral TNF plays a crucial role in the antitumor mechanisms of DT-5461 and suggest that its antitumor action is influenced by an augmentative effect of tumor cells on TNF production and by blood levels of immunosuppressive factors. PMID- 9412658 TI - Conformational and electrostatic properties of naphthazarin, juglone, and naphthoquinone: an ab initio theoretical study. AB - Conformational features of naphthazarin, juglone, and naphthoquinone have been examined via ab initio (Hartree-Fock) SCF calculations at 3-21G level. The results suggest a planar structure for all the three molecules and internally hydrogen-bonded structure for naphthazarin and juglone to be their preferred conformation. The optimized structural features are essentially the same as their crystal geometries. Molecular electrostatic potential (MEP) calculations using ab initio SCF methods ranging from 3-21G to 6-31G levels have been performed to visualize their three-dimensional pharmacophoric patterns and topography. The results indicate that two factors--(i) the depth, extent, and relative location of negative potential around hydroxyl and quinonoid oxygens, and (ii) a gradual loss of negative potential over the molecular plane due to the presence and orientation of the hydroxyl groups in the phenolic part of the molecules--are crucial for recognition interaction of the compounds with their receptors. Aqueous solvation seems to have significant influence on the MEP profiles of the molecules. Although intrinsic nucleophilicity increases for all the compounds, including the different conformers, due to aqueous solvation, the intrinsic electrophilicity shows remarkable decrease for all. It appears that the acidic nature of the hydrogens in these compounds and conformers decreases sharply along with shifts of positions while going from the gas phase to the aqueous phase. These observations may help to explain the mechanism of action(s) of the anthracyclin family of cytotoxic antibiotics. PMID- 9412660 TI - Introduction to genetic linkage analysis. AB - Advances in molecular methodology have resulted in the successful genetic mapping and isolation of the genes responsible for numerous simple Mendelian disorders such as cystic fibrosis, neurofibromatosis, and Huntington disease. Recently, researchers have begun to successfully study diseases with more complex inheritance, such as breast cancer. This paper will provide a basic introduction to the strategies and methods commonly used in the localization of genes resulting in disease. PMID- 9412659 TI - A pharmacodynamic study of morphine and its glucuronide metabolites after single morphine dosing in cancer patients with pain. AB - Eleven morphine naive patients with cancer-related pain were given a single dose of either intravenous morphine (n = 5) or oral morphine (n = 6). Blood sampling was performed over a 24-hr period and serial pain assessments were made using a categorical scale. Plasma samples were analyzed for morphine, morphine-6 glucuronide (M-6-G), morphine-3-glucuronide (M-3-G), and normorphine using high performance liquid chromatography. In neither the intravenous nor oral group was there a correlation between analgesia duration and the half-lives of morphine and M-6-G. There was no correlation between the time to peak analgesia and time to peak concentration for morphine or M-6-G. There was no significant difference in absolute concentrations of M-6-G or M-3-6 nor in the ratio of M-3-G to M-6-G at peak analgesia versus relapse. PMID- 9412661 TI - Advances in autologous stem cell transplantation. PMID- 9412662 TI - A clinical review of the evidence for the role of ultraviolet radiation in the etiology of cutaneous melanoma. AB - Strong epidemiological evidence exists that solar radiation is causally related to a significant proportion of cutaneous melanoma. The nature of the relationship is, however, complex and the details are not entirely clear. There appears to be a complex interplay between solar exposure in individuals with a characteristic phenotype. Although the exact quantitative and qualitative nature of this exposure is not clear, it is probable that intermittent exposures and intense exposures with consequent sunburns in a high-risk phenotype are critical in increasing the risk of developing melanoma. Despite the lack of complete understanding of this relationship, the evidence is convincing that solar radiation is causally related to cutaneous melanoma and consequently exposures to UV radiation should be reduced from early in life. The preventive measures involve reducing exposure to ambient solar radiation, by avoiding peak exposures, wearing protective outerwear, and using broad-spectrum sunscreens. PMID- 9412663 TI - Breast cancer: cell and gene therapy. PMID- 9412664 TI - Development of vaccine strategies for the treatment of B-cell malignancies. PMID- 9412666 TI - Fractals, chaos, and cancer: do they coincide? PMID- 9412665 TI - The chloroethylnitrosoureas: sensitivity and resistance to cancer chemotherapy at the molecular level. AB - The chloroethylnitrosoureas were developed in a synthetic program that began with the observation that N-methyl-N'-nitro-N-nitrosoguanidine was an effective agent against L1210 cells. The antitumor activity of the chloroethylnitrosoureas is based on their reactions with DNA, especially the formation of a cytosine-guanine crosslink in DNA. Resistance occurs when the enzyme, O6-alkylguanine-DNA alkyltransferase, repairs an intermediate in crosslink formation. Inhibition of O6-alkylguanine-DNA alkyltransferase often restores sensitivity to the chloroethlylnitrosoureas although evidence is accumulating that other repair mechanisms may also contribute to the resistance phenomenon. Continuing investigations in this field center on finding agents whose reactions with DNA are more specific, on elucidating other resistance mechanisms, and on overcoming resistance by developing new inhibitors of repair enzymes. PMID- 9412667 TI - Alice and academic oncologists meet in corporate wonderland. PMID- 9412668 TI - Dose-volume histograms can be interpreted in different ways. PMID- 9412669 TI - The emerging role of neoadjuvant hormonal therapy in the management of localized prostatic cancer--reply. PMID- 9412670 TI - Spay Day USA: controversy eclipses mutual pursuit. Interview by Susan C. Kahler. PMID- 9412671 TI - Is cloning necessary? PMID- 9412672 TI - Efficacy of low dosages of epoetin alfa in dogs. PMID- 9412674 TI - Improving the ability of food animal veterinarians to make management decisions. PMID- 9412673 TI - Opposes manufacturer's policy on sale of antiflea product. PMID- 9412675 TI - What is your diagnosis? Osteolytic lesion involving the distal portion of the diaphysis of the left femur: osteosarcoma. PMID- 9412676 TI - Importance of companion animals in children's lives--implications for veterinary practice. PMID- 9412677 TI - Employment, starting salaries, and educational indebtedness of 1997 graduates of US veterinary medical colleges. PMID- 9412678 TI - Attitudes of veterinarians, animal control directors, and county prosecutors in Michigan regarding enforcement of state animal cruelty legislation. AB - OBJECTIVES: To determine attitudes of veterinarians, animal control directors, and country prosecutors in Michigan toward enforcement of state animal cruelty legislation and to identify factors associated with whether veterinarians would report suspected cases of animal cruelty. DESIGN: Survey. SAMPLE POPULATION: Questionnaires were sent to 1,146 Michigan Veterinary Medical Association member veterinarians, 139 animal control directors, and 83 county prosecutors in Michigan. RESULTS: 740 (65%) veterinarians, 70 (50%) animal control directors, and 43 (52%) prosecutors responded. Six hundred forty six of 735 (88%) veterinarians reported having treated an animal that they believed had been a victim of animal cruelty, but only 192 of 719 (27%) had ever reported a case of animal cruelty, and only 217 of 734 (30%) had ever testified in an animal cruelty case. Logistic regression analysis of responses revealed that the only factor associated with whether veterinarians would report cases of suspected animal cruelty was the potential reactions of the involved clients to the accusation of animal cruelty. Veterinarians who rated reaction of the involved client as important, very important, or essential to their decision whether to report a case of animal cruelty were less likely to report such cases than were veterinarians who rated potential client reaction as somewhat important or unimportant. CLINICAL IMPLICATIONS: Concern about potential client reaction was the most important factor in whether veterinarians would report cases of suspected animal cruelty. PMID- 9412679 TI - Rabies surveillance in the United States during 1996. AB - In 1996, 49 states, the District of Columbia, and Puerto Rico reported 7,124 cases of rabies in non-human animals and 4 cases in human beings to the Centers for Disease Control and Prevention. Nearly 92% (6,550 cases) were wild animals, whereas 8% (574 cases) were domestic species. The total number of reported cases decreased 9.6% from that of 1995 (7,881 cases). Although much of the decline was the result of fewer reported cases of rabies in raccoons, fewer cases were also reported among most groups of animals. Numbers of cases associated with separate epizootics of rabies in foxes in west central Texas and in dogs and coyotes in southern Texas attributable to canine variants have declined, with 56.2% fewer rabid foxes (60), 72.7% fewer rabid dogs (15), and 76.3% fewer rabid coyotes (19) during 1996, compared with cases of rabies reported among these same species during 1995. Nationally, the number of reported rabid bats (741) decreased 5.8%, with cases reported by 46 of the 48 contiguous states. Four Eastern Seaboard states, enzootic for the raccoon variant of the rabies virus, reported noteworthy increases in total numbers of reported cases: Maine (29.7%; 101 cases in 1995 to 131 in 1996), Maryland (44.2%; 441 to 636), North Carolina (59.0%; 466 to 741), and Virginia (33.3%; 459 to 612). Increases were also reported by Florida (6.4%; 251 to 267) and Georgia (3.1%; 294 to 303). Cats continued to be the domestic animal most frequently reported rabid, but reported cases of rabies in cats (266), cattle (131), and dogs (111) decreased by 7.6%, 3.7%, and 24.0%, respectively. Thirty-one states and the District of Columbia reported decreases in rabies in animals during 1996, compared with 18 states and Puerto Rico in 1995. Hawaii was the only state that did not report a case of rabies in 1996. Two indigenously acquired cases of rabies reported in human beings were the result of infection with rabies virus variants associated with bats, whereas the remaining 2 human rabies infections were acquired outside the United States, and the variants identified were consistent with those associated with rabid dogs. PMID- 9412681 TI - Surgical management of extrathoracic tracheal collapse in two large-breed dogs. AB - Collapse of the cervical portion of the trachea was diagnosed for 2 young large breed dogs. Clinical signs included worsening respiratory stridor and coughing exacerbated by exercise. The diagnosis was confirmed by use of conventional radiography and fluoroscopy of the trachea. A polypropylene, spiral ring, extraluminal, tracheal prosthesis was used to successfully treat tracheal collapse in each dog. Although tracheal collapse typically affects middle-aged and old small-breed dogs, tracheal collapse should be considered as a differential diagnosis for large-breed dogs with a honking cough and respiratory stridor. PMID- 9412680 TI - Retrospective cohort study of changes in hip joint phenotype of dogs in the United States. AB - OBJECTIVE: To determine whether there had been a significant improvement in hip joint phenotype of dogs in the United States by comparing results of evaluations done by the Orthopedic Foundation for Animals of dogs born between 1972 and 1980 with those of dogs born between 1989 and 1992 and determining whether there had been an increase in the percentage of dogs classified as having excellent hip joint phenotype. DESIGN: Retrospective cohort study. SAMPLE POPULATION: 270,978 evaluations. PROCEDURE: Numbers and percentages of dogs classified as having excellent hip joint phenotype during each period and change between periods in percentages of dogs classified as having excellent hip joint phenotype were calculated. RESULTS: Percentage of dogs born between 1989 and 1992 that were classified as having excellent hip joint phenotype (15,289/143,668; 10.64%) was significantly higher than percentage of dogs born between 1972 and 1980 that were classified as having excellent hip joint phenotype (9,960/127,310; 7.82%). The increase in percentage of dogs classified as having excellent hip joint phenotype was significantly higher for male (51%) than for female (27%) dogs. CLINICAL IMPLICATIONS: Results suggest that there has been an improvement in the hip joint phenotype of dogs in the United States between the 1970s and early 1990s and that the improvement has been greater among male than among female dogs. PMID- 9412682 TI - Removal of a cholesteatoma in a dog, using a caudal auricular approach. AB - A new surgical method for treating cholesteatoma in dogs is described. Although rarely reported in dogs, cholesteatomas may be more common than previously believed. Complete excision of a cholesteatoma is associated with low recurrence and good long-term prognosis. Surgical intervention, with total ear canal ablation and lateral bulla osteotomy, has been recommended in dogs with tumors of the middle ear; however, this technique often results in conductive hearing loss. Through a caudal auricular approach to the tympanic bulla, we were able to preserve the external ear canal, reconstruct the auditory ossicles, and graft the tympanic membrane. Results of brain stem auditory-evoked response tests in the dog revealed intact conduction potentials. A caudal auricular approach to the tympanic bulla is technically possible, preserves normal appearance, and may maintain, or even improve, hearing conduction of affected ears in dogs. PMID- 9412683 TI - Outbreak of Salmonella infantis infection in a large animal veterinary teaching hospital. AB - During the past 11 years, there have been numerous reports of outbreaks of salmonellosis involving horses in veterinary teaching hospitals. Some of these outbreaks have been associated with Salmonella serotypes not commonly associated with infection of horses. Salmonella infantis is among the more common Salmonella serotypes isolated from human beings, and is an important pathogen in the broiler chicken industry. However, it was not commonly isolated from horses or cattle on a national basis between 1993 and 1995. In this report, we describe an outbreak of S infantis infection among large animals, primarily horses, in a veterinary teaching hospital and the control measures that were implemented. Factors that appeared to be key in control of this outbreak in this hospital included providing biosecurity training sessions for hospital personnel, adopting a standard operating procedure manual for biosecurity procedures, installing additional handwashing sinks throughout the facility, painting the interior of the facility with a nontoxic readily cleanable paint, replacing the dirt flooring in 4 stalls with concrete flooring, and removing noncleanable surfaces such as rubber stall mats, wooden hay storage bins, and open grain bins. Our experience with this outbreak suggests that although it is virtually impossible to eliminate Salmonella organisms from the environment, minimizing contamination is possible. Prevention of nosocomial infection must be approached in a multifaceted manner and care must be taken to search out covert sources of contamination, especially if standard intervention procedures do not prevent spread of the disease. PMID- 9412684 TI - Abscess on the lateral epicondyle of the humerus as a cause of lameness in a horse. AB - An 18-month-old 450-kg [990-lb] sexually intact male Holsteiner was evaluated for lameness of the left forelimb of 3-months' duration. We were unable to localize the site of lameness, using intra-articular and perineural anesthesia, and radiography of the shoulder and cubital joint (elbow) did not reveal radiographic abnormalities. Nuclear scintigraphy was performed. An increase in radio-isotope uptake was evident at the lateral epicondyle of the left humerus. Radiographs of the region 3 weeks later revealed a 1.5-cm focal lucency surrounded by a 1.0-cm rim of necrotic bone. The lesion was consistent with an encapsulated osteomyelitis or bone abscess. Surgical correction was performed, using general anesthesia, and involved a lateral approach to the epicondyle of he humerus. Cyst contents were removed with curettage, and the cavity was packed with cancellous bone harvested from the tuber coxae. A coagulase-negative Staphylococcus organism was cultured from the abscess. The horse was sound 3 weeks after surgery, and radiography 10 months later revealed complete ossification and adjacent sclerosis at the surgical site. Solitary cystic lesions of long bones may represent bone abscesses capable of causing lameness in horses. Nuclear scintigraphy can provide early diagnostic capability. Curettage and cancellous bone grafting are indicated for treatment of bone abscesses. PMID- 9412686 TI - Clinical and radiographic findings, treatment, and outcome in cattle with osteochondrosis: 29 cases (1986-1996). AB - OBJECTIVE: To summarize the radiographic and clinical findings, treatment, and outcome in cattle with osteochondrosis diagnosed radiographically. DESIGN: Retrospective case series. SAMPLE POPULATION: 29 cattle with radiographic evidence of osteochondrosis. PROCEDURES: Medical records were reviewed, and owners or referring veterinarians were contacted for outcome assessment. Data were analyzed for potential interactions between osteochondrosis classification (osteochondritis dessicans vs subchondral cyst-like lesions), clinical and radiographic findings, treatment, and outcome, using Fisher's exact test and descriptive statistics. RESULTS: Osteochondrosis was associated with young, male, purebred cattle, clinical evidence of lameness, and radiographic evidence of concurrent degenerative joint disease. Osteochondritis dissecans and subchondral cyst-like lesions had similar clinical findings and outcomes but varied significantly in their radiographic distribution among joints. Osteochondrosis often manifests clinically as a unilateral condition, but bilateral lesions were often found (88%) when limbs were radiographically examined. Cattle managed conservatively tended to be culled (within 6 months of diagnosis because of lameness) more often than those managed surgically, despite the lack of treatment bias. CLINICAL IMPLICATIONS: Osteochondrosis in cattle is often associated with lameness or degenerative joint disease. Conservative management does not result in a favorable clinical prognosis for long-term, lameness-free survival, and more studies need to be completed to evaluate the efficacy of surgical treatment of osteochondrosis in cattle. PMID- 9412685 TI - Sedative effects of medetomidine and its reversal by atipamezole in llamas. AB - OBJECTIVE: To determine a dose of medetomidine that will induce sedation in llamas, to assess effects of medetomidine sedation on arterial blood gas variables, and to determine efficacy of atipamezole in reversing medetomidine induced sedation. DESIGN: Prospective, randomized clinical trial. ANIMALS: 15 clinically normal adult llamas. PROCEDURE: 9 llamas received various doses of medetomidine (0.01, 0.02, or 0.03 mg/kg [0.005, 0.009, or 0.014 mg/lb] of body weight, i.m.). Heart and respiratory rates and sedative effects were recorded. Using the lowest dose that induced deep sedation, 6 different llamas were used to assess effects of medetomidine on arterial blood gas variables. These same 6 llamas were later given atipamezole (0.125 mg/kg [0.057 mg/lb], i.v.) 30 minutes after medetomidine injection. Heart and respiratory rates, sedative effects, and time from atipamezole injection to standing were recorded. RESULTS: Sedation began 6.67 +/- 1.15 minutes (mean +/- SD) after medetomidine administration (0.03 mg/kg, i.m.). Arterial blood gas variables measured 30 and 60 minutes after injection were not different from baseline. Llamas that did not receive atipamezole remained recumbent for 91.50 +/- 24.68 minutes. After atipamezole administration, llamas were able to stand in 5.80 +/- 3.27 minutes. CLINICAL IMPLICATIONS: Medetomidine induced light to deep sedation in a dose-dependent manner in clinically normal llamas. A dose of 0.03 mg/kg induced deep sedation with a short period of analgesia. Atipamezole rapidly reversed effects of medetomidine, and llamas recovered quickly and were soon able to stand. PMID- 9412687 TI - Paraintestinal mesenteric abscess and chronic peritonitis in a bull. AB - A 2-year-old bull was examined because of intermittent anorexia, signs of mild colic, and weight loss of 3 weeks' duration. A tympanitic resonance (ping) could be heard during simultaneous auscultation and percussion of the right paralumbar fossa, and a mass could be felt in the right dorsal quadrant of the abdominal cavity during palpation per rectum. Right flank laparotomy was performed, and intraoperative ultrasonography and ultrasound-guided fine-needle aspiration were used to determine that the mass was an abscess. However, the abscess could not be removed or drained into the colon because of extensive adhesions to other organs. Because the owner refused to pursue continued medical treatment, the bull was euthanatized. At necropsy, the abscess was found to be connected to a caudal mesenteric lymph node through a fistula. Histologic evaluation of the lymph node revealed hyperplastic lymphadenitis, and an alpha-hemolytic streptococcus was recovered from the abscess fluid. The most likely possibility for the findings in this bull were that the lymphadenitis was of hematogenous origin and that the abscess developed as a direct extension of the infectious process, similar to development of mesenteric abscesses in horses with chronic streptococcal infection (i.e., strangles). PMID- 9412688 TI - Survey of diplomates of the American College of Veterinary Surgeons regarding postoperative intra-abdominal adhesion formation in horses undergoing abdominal surgery. AB - OBJECTIVE: To obtain information from specialists in equine surgery as to prevalence of, predisposing factors for, and methods to prevent postoperative adhesion formation in horses undergoing abdominal surgery. DESIGN: Survey. PROCEDURE: Surveys were mailed to 196 diplomates of the American College of Veterinary Surgeons involved in equine practice. RESULTS: 60 (31%) surveys were returned. Most respondents (55/60) routinely informed clients of the risk of postoperative adhesion formation in horses with small intestinal lesions. When asked after which procedures they routinely used measures to prevent adhesions, 56 of 60 (93%) indicated that they did after small intestinal resection and anastomosis and 56 of 60 (93%) indicated that they did after any abdominal surgery in foals. The 4 methods most frequently listed when respondents were asked which methods were effective at preventing adhesion formation were meticulous surgical technique, administration of antibiotics and nonsteroidal anti-inflammatory drugs, intraoperative peritoneal lavage, and methods that prevent abdominal contamination. Most respondents (50/60) thought that at least some horses with colic secondary to adhesion formation could be managed medically. Fifty-four (90%) respondents indicated that they were successful less than half of the time when treating horses with adhesions severe enough to require additional surgery. CONCLUSION: In general, respondents thought that less than 15% of horses undergoing abdominal surgery would develop adhesions, but that horses with small intestinal disease and foals were most prone to develop adhesions. Meticulous surgical technique was thought to be the most important factor in preventing adhesions, and many prevention regimens reported to be effective in the literature were not commonly used in practice. PMID- 9412689 TI - On being an academic radiologist in a small department, or bigger ain't always necessarily better. PMID- 9412691 TI - Development of a radiology faculty appraisal instrument by using critical incident interviewing. AB - RATIONALE AND OBJECTIVES: To develop a valid and reliable radiology faculty appraisal instrument based on scientific methods. MATERIALS AND METHODS: Fifteen radiology residents participated in critical incident interviewing. During a 1 hour interview, a resident was asked to describe five incidents each of effective and ineffective faculty behavior. Two investigators independently listened to the tape-recorded interviews, and two different investigators sorted the incidents into broad categories. A faculty appraisal instrument was developed by listing similar incidents under broad categories. A five-point rating scale was applied to each item. Content validity was assessed by resident and faculty critique of the appraisal instrument. RESULTS: A total of 168 incidents of faculty behavior were generated. The frequency with which similar incidents were reported was recorded. The most common behaviors reported were related to staff expertise and teaching. Interjudge reliability was good, as determined by computing K indices of agreement (overall K = 0.59). There was good agreement regarding instrument content validity among residents but not among faculty. CONCLUSION: Residents supported the use of the new appraisal instrument, but further tests of validity and reliability and faculty acceptance of the instrument will determine its usefulness as a tool for monitoring faculty teaching performance and making decisions regarding faculty promotion. PMID- 9412690 TI - Attenuation of contrast material-induced renal artery vasoconstriction by nitric oxide donors. AB - RATIONALE AND OBJECTIVES: The authors studied the role of the endothelium and associated endothelial pathways in contrast material-induced renal vasoconstriction. MATERIALS AND METHODS: Isometric contractions in human and rabbit renal artery rings with intact and denuded endothelium were stimulated with phenylephrine and increasing concentrations of the ionic contrast material diatrizoate, the nonionic contrast materials iopamidol and iomeprol, and the dimeric contrast material iodixanol in a tissue perfusion bath. Rings with intact endothelium were incubated with endothelium-stimulating compounds such as the NO synthetase inhibitor Ng nitro-L-arginine methyl ester (L-NAME) to study the endothelium-mediated vasomotor regulation and the NO-liberating substances molsidomine (SIN-1) and nitroprusside (NPR) to study the endothelial-mediated vasorelaxation before being stimulated with contrast material. RESULTS: Contrast material-induced, dose-dependent, reversible renal artery contractions are dependent on the type of contrast material. No differences in the contractions were found between intact and denuded rings. L-NAME had no effect on contrast material-induced contractions. Contractions were inhibited by the NO donors SIN-1 and NPR. SIN-1 was the most potent inhibitor. CONCLUSION: Contrast material induced renal vasoconstriction is endothelium-independent. Selective pharmacologic stimulation of the endothelium by NO donors, however, may still be useful in the prophylaxis of contrast material-induced renal vasoconstriction and, thus, potentially nephrotoxicity. PMID- 9412692 TI - Outcome of examinations self-referred as a result of spiral CT of the abdomen. AB - RATIONALE AND OBJECTIVES: The interpretation of an abdominal computed tomographic (CT) scan is occasionally inconclusive. In many of these cases, the radiologist suggests an additional imaging test for further confirmation or clarification. The purpose of this study was to evaluate the outcome of self-referral by the radiologist after abdominal CT scanning. MATERIALS AND METHODS: Reports from 545 consecutive abdominal CT scans were reviewed to track recommendations for additional imaging. In patients who underwent the additional work-up, a determination of the effect of the study was attempted. In patients who did not, explanations were sought. Wording of the recommendations was also recorded. RESULTS: Recommendations were made for additional imaging studies in 105 (19.3%) patients. Of these, 32 (30.5%) were performed and 31 (96.9%) were helpful by confirming malignancy (n = 5), confirming a benign process (n = 24), or being therapeutic (n = 2). In one, no information was added. There were 63 (60.0%) patients who did not undergo the recommended studies. Reasons included "no clinical indication" (n = 51), alternative study performed (n = 9), or study previously performed (n = 3). In eight (7.6%) patients the chart provided insufficient information about whether the patient underwent the study, and in two (1.9%) the chart was unavailable. Wording of the recommendation had no effect on whether the study was performed (P > .05). CONCLUSION: Although interpretation of abdominal CT scans leads to recommendations for additional imaging in a minority of cases, these recommendations were infrequently followed. When followed, however, the findings from the recommended studies were usually helpful. Better clinical information is perhaps the best way to reduce self referral by radiologists. PMID- 9412693 TI - Work-up of patients with malignancy after an intermediate-probability ventilation perfusion scan: why don't physicians pursue a definitive diagnosis? AB - RATIONALE AND OBJECTIVES: The authors determined whether there are specific patient characteristics associated with the clinical decision to eschew further diagnostic testing in patients in whom a ventilation-perfusion (V-P) scan indicates intermediate probability of pulmonary embolism (PE). MATERIALS AND METHODS: The authors reviewed all intermediate-probability V-P scans obtained in a 12-month period. Patients were divided into two groups. Group 1 comprised patients in whom a definitive diagnosis of PE was not obtained and who were not treated for PE (n = 57); group 2 comprised patients in whom the diagnosis of PE was confirmed or excluded and who, if PE was confirmed, received appropriate treatment (n = 14). Age, gender, frequency of malignancy, and survival of patients in groups 1 and 2 were compared. RESULTS: The frequency of malignancy was significantly greater in group 1 than in group 2 (P = .012). Although the estimated 2-year survival of group 1 patients was significantly less than that of group 2 patients (P = .039), this difference is likely due to confounding by age and malignancy. CONCLUSION: When an intermediate-probability V-P scan is obtained, physicians are significantly less likely to pursue a definitive diagnosis of PE in patients with malignancy. PMID- 9412694 TI - Magnetization transfer imaging of bone marrow with and without fat suppression. AB - RATIONALE AND OBJECTIVES: To assess magnetization transfer (MT) ratios of bone marrow at magnetic resonance (MR) imaging performed with and without fat suppression. MATERIALS AND METHODS: The authors performed MR imaging in 30 regions of normal bone marrow in 10 subjects by using four types of gradient-echo sequences: a combination of MT and fat-suppression techniques, only the fat suppression technique, only the MT technique, and without MT or fat-suppression techniques. MT ratios of marrow obtained with and without the fat-suppression technique were quantitatively compared. RESULTS: The average MT ratio of marrow with fat suppression was significantly higher than that without fat suppression (P < .01). CONCLUSION: Because bone marrow includes both water and fat, the MT ratio of marrow was underestimated when MT imaging was performed without fat suppression. A fat-suppression technique should be used. PMID- 9412695 TI - Saquinavir pharmacokinetics alone and in combination with nelfinavir in HIV infected patients. AB - OBJECTIVE: To investigate the pharmacokinetics of saquinavir (SQV) hard gel when administered alone and in combination with nelfinavir (NLF) to HIV-positive patients. DESIGN: Six patients receiving triple therapy (dual nucleoside plus SQV 600 mg three times daily) were studied. On the first study day blood samples were drawn for assay of SQV. Prior to the second study day, patients received their usual medication plus NLF 750 mg three times daily for 2 days. METHODS: Blood samples were obtained at times 0, 1, 2, 4, 6 and 8 h after dosing on study days 1 and 2. Following centrifugation, separated plasma was heated at 58 degrees C for at least 30 min to inactivate HIV and stored at -80 degrees C until analysis using high performance liquid chromatography. RESULTS: The geometric mean Cmax and AUC0-8 h on the first study day were 253 ng/ml (range, < 25-1200 ng/ml) and 1106 ng/ml.h (range, < 100-3479 ng/ml.h), respectively, and on the second study day were 1204 ng/ml (range, 379-2755 ng/ml) and 5472 ng/ml.h (range, 1434-12,538 ng/ml.h), respectively. The geometric mean ratio for Cmax was 4.75 and for AUC0-8 h was 4.94. CONCLUSIONS: NLF increases the oral bioavailability of SQV (hard gel) approximately fivefold. For some patients the addition of NLF to SQV will increase the drug levels from subtherapeutic to the therapeutic range. In one of our patients the addition of NLF resulted in SQV levels that were much higher than previous work suggests are necessary for maximum antiviral effect. The variability in SQV concentrations both at baseline and following addition of NLF suggest that dosing may best be adjusted by individual therapeutic drug monitoring. PMID- 9412696 TI - Cytomegalovirus disease in AIDS. PMID- 9412697 TI - AIDS prognosis based on HIV-1 RNA, CD4+ T-cell count and function: markers with reciprocal predictive value over time after seroconversion. AB - OBJECTIVE: HIV-1 RNA levels in peripheral blood are strongly associated with progression to AIDS, CD4+ T-cell decline, or death. Their predictive value is reportedly independent of the predictive value of CD4+ T-cell counts. Because the interrelations between these parameters of HIV-1 infection are poorly understood, we studied the kinetics and predictive value of serum HIV-1 RNA levels, CD4+ T cell counts, and T-cell function. DESIGN AND METHODS: HIV RNA levels, CD4+ T-cell counts, and T-cell function were measured from seroconversion to AIDS in 123 homosexual men who seroconverted during a prospective study and were followed over 10 years. RESULTS: Two patterns of median HIV-1 RNA levels were found during infection: a steady-state and a 'U-shaped' curve. Steady-state high RNA levels were related to rapid disease progression. For the U-shaped curve, there were groups with high and low RNA levels related to disease progression. At 1 year after seroconversion, RNA level was the only marker that was strongly predictive. Furthermore, decreasing RNA levels in the first year following seroconversion were related to better prognosis than stable low levels. Low CD4+ T-cell count and T-cell function became predictive of progression to AIDS at 2 and 5 years after seroconversion, respectively. CONCLUSIONS: With ongoing infection, the predictive value of low CD4+ T-cell count and T-cell function increases, whereas the predictive value of high HIV-1 RNA level decreases. These findings reflect the observation that infection with HIV progressively leads towards immune deficiency, which in later stages is most predictive of disease progression. PMID- 9412698 TI - gp120-directed antibody-dependent cellular cytotoxicity as a major determinant of the rate of decline in CD4 percentage in HIV-1 disease. AB - OBJECTIVE: To determine the relationship between the rate of CD4 percentage decline and two factors postulated to be associated with CD4 cell destruction: circulating HIV-1 viral load and gp120-directed antibody-dependent cellular cytotoxicity (ADCC). DESIGN: Four women and 16 men had serial determinations of CD4 percentage gp120-directed ADCC activity [using the cell-mediated cytotoxicity (CMC) assay] natural killer (NK) cell number, spontaneous NK lytic function, and plasma HIV-1 RNA. METHODS: The rate of decline in CD4 percentage was modeled as a function of gp120-directed ADCC activity and circulating HIV-1 RNA using Pearson correlation and multiple regression analyses. RESULTS: All individuals had at least four CMC assays performed and two HIV-1 RNA polymerase chain reaction measurements over a median follow-up of 27 months. Although the rate of CD4 percentage decline was associated with either CMC activity (r = -0.53, P = 0.02) or circulating HIV-1 RNA (r = -0.42, P = 0.07), it was strongly correlated with an interaction between CMC and HIV-1 RNA (r = -0.76, P < 0.0001). Mean CMC activity was associated with both mean percentage of circulating NK cells and mean spontaneous NK cell lysis. CONCLUSIONS: The ability of cells from HIV infected individuals to mediate gp120-directed ADCC, together with a sufficient circulating viral load, define conditions under which rapid CD4 cell destruction may occur. This relationship between viral load and an HIV-1-specific immune response lends important insights into the central causes of immunodeficiency in AIDS and suggests additional avenues for therapeutic intervention. PMID- 9412699 TI - Field evaluation of alternative testing strategies for diagnosis and differentiation of HIV-1 and HIV-2 infections in an HIV-1 and HIV-2-prevalent area. AB - OBJECTIVE: To identify cost-efficient alternative antibody testing strategies for screening, confirmation and discrimination of HIV-1 and HIV-2 infections, including rapid simple tests (RST) as well as enzyme-linked immunosorbent assays (ELISA), in a HIV-1 and HIV-2-prevalent area. DESIGN: Evaluation and comparison of anti-HIV-1/2 assays, adhering to the World Health Organization recommendations for alternative confirmatory strategies, using banked and prospectively collected specimens in Guinea-Bissau. METHODS: A total of 1110 consecutive sera from Bissau were included in the first phase, of which 198 (17.8%) were HIV-seropositive: 52 (4.7%) HIV-1, 120 (10.8%) HIV-2, and 26 (2.3%) HIV-1/HIV-2 dually reactive. In addition, 95 selected HIV-positive specimens were included for study of sensitivity and cross-reactivity between HIV-1 and HIV-2. Western blot was used as a gold standard for confirming the reactivity of the specimens. All specimens were screened by two assays. Enzygnost ELISA and Capillus RST. Samples reactive by any of the screening assays were further tested by assays chosen for confirmation: UBI ELISA, Innotest ELISA Recombigen RST, Multispot RST and Immunocomb RST. The confirmatory RST as well as Wellcozyme Recombinant HIV-1 ELISA, PEPTI-LAV and INNO-LIA were also used to study differentiation between HIV 1 and HIV-2. RESULTS: The sensitivities of all assays were 100%. The specificities of the screening assays at initial and repeated testing were 98.0 and 99.7%, respectively, for Enzygnost and 99.8 and 99.9%, respectively, for Capillus. The various combinations of two or three assays showed specificities of 99.2-100%. Several possible combinations of assays were identified where a specificity of 100% and good differentiation between HIV-1 and HIV-2 was achieved. Significant differences in the capacity to discriminate were noted; Immunocomb and PEPTI-LAV had the lowest number of dual-reactive results. A follow up study of 1501 consecutive samples tested with the strategy chosen for routine use showed a sensitivity and specificity comparable to ELISA and Western blot. CONCLUSION: High sensitivities and specificities were obtained with various combinations of assays including RST as well as ELISA, and these procedures are well suited for field use in Africa. Serodiagnostic strategies for HIV can be based on RST alone and differentiation between HIV-1 and HIV-2 can be achieved as part of these strategies. Large differences in the capacity of individual assays to discriminate between HIV-1 and HIV-2 were observed. PMID- 9412702 TI - Clinically significant azole cross-resistance in Candida isolates from HIV positive patients with oral candidosis. AB - OBJECTIVES: To determine the proportion of fluconazole-resistant Candida albicans isolates that have clinically significant cross-resistance to itraconazole or ketoconazole, that is sufficient to result in failure of these agents at their standard doses (200 and 400 mg daily for 7 days, respectively). METHODS: Seven hundred C. albicans isolates from HIV-positive patients with oral candidosis underwent susceptibility testing using a relative growth method, for which cut off values corresponding to clinical drug failure have been established. RESULTS: A total of 431 isolates were fully azole-susceptible and three main resistance patterns were detected: isolates resistant to fluconazole alone (n = 100); isolates resistant to fluconazole and ketoconazole but susceptible to itraconazole (n = 94); and isolates resistant to all three drugs (n = 50). No isolates were consistently resistant to ketoconazole without being fluconazole resistant, and no itraconazole resistance was detected without ketoconazole resistance. Resistance to fluconazole alone was more common in specimens obtained soon after first clinical fluconazole failure, whereas specimens from patients with a longer history of fluconazole-unresponsive candidosis were more likely to be infected with cross-resistant isolates. Median days of prior azole exposure and cumulative fluconazole dose were significantly less for those with isolates resistant to fluconazole alone than for those with ketoconazole cross-resistant isolates, who had received less azole therapy and smaller cumulative fluconazole doses than those with isolates cross-resistant to all three drugs (although not statistically significant). After the diagnosis of fluconazole-unresponsive candidosis, increasing cumulative doses of itraconazole solution were associated with increasing likelihood of cross-resistance. CONCLUSIONS: Clinically significant cross-resistance to other azoles may occur in fluconazole-resistant isolates of C. albicans, although initially most isolates are not cross-resistant and the detection of cross-resistant isolates is associated with a history of greater prior azole exposure. Patients who have been treated for fluconazole resistant candidosis for longer and with greater cumulative doses of itraconazole solution tend to become infected with increasingly cross-resistant isolates of C. albicans. PMID- 9412701 TI - Increased frequency of CCR-5 delta 32 heterozygotes among long-term non progressors with HIV-1 infection. The Australian Long-Term Non-Progressor Study Group. AB - BACKGROUND: The beta-chemokine receptor CCR-5 is used as a coreceptor by macrophage-tropic strains of HIV-1 to gain entry into CD4+ cells. OBJECTIVE: To determine the effect of a common 32 base-pair deletion mutation in the CCR-5 gene (CCR-5 delta 32) on progression of HIV infection to AIDS, and to assess the level of heterozygosity for this mutation in a well-defined group of long-term non progressors (LTNP). PARTICIPANTS: Sixty-four HIV-1-infected LTNP (CD4+ T lymphocyte count > 500 x 10(6)/l after 8 years) were compared with 95 individuals infected within a similar period (1983-1986) but who had rapidly progressed to AIDS and death, and with a further 120 HIV-positive individuals with CD4+ counts < 500 x 10(6)/l. METHODS: The presence of the CCR-5 delta 32 mutation was assessed using polymerase chain reaction with primers spanning the 32 base-pair deletion. CD4+ and CD8+ counts, plasma HIV-1 RNA, p24 antigen and beta 2 microglobulin levels in LTNP carrying the CCR-5 delta 32 mutation were compared with LTNP lacking the mutation. RESULTS: A marked increase in the frequency of CCR-5 delta 32 heterozygosity was found among LTNP (35.9%) compared with rapid progressors (12.6%; P = 0.0005) and patients selected on the basis of a CD4+ T cell count < 500 x 10(6)/l (12.5%; P = 0.0004). LTNP heterozygous for CCR-5 delta 32 had a significantly higher CD8+ T-cell count than those without the mutation (1218 versus 972 x 10(6)/l; P = 0.044). No significant correlation was observed between heterozygosity and CD4 count, viral load, p24 antigen or beta 2 microglobulin within the LTNP group. CONCLUSIONS: This study provides the strongest evidence to date for the importance of a single copy of the CCR-5 delta 32 mutation in long-term non-progression of HIV infection, which may involve, in part, CD8+ T lymphocytes. PMID- 9412700 TI - Serologic and phylogenetic characterization of HIV-1 subtypes in Uganda. AB - OBJECTIVES: To determine the HIV genetic subtypes present in HIV-1-infected asymptomatic blood donors in Uganda and to evaluate serologic detection of infection by commercial immunoassays; to evaluate samples for HIV-1 group O infections. METHODS: Sixty-four HIV-seropositive plasma samples were collected from the Nakasero Blood Bank, Kampala, Uganda. The plasma were evaluated using commercial HIV enzyme immunoassays (EIA) and a research immunoblot. HIV-1 group M and O infections were identified on the basis of discordant seroreactivity in EIA and reactivity to group M and O antigens on the immunoblot. Regions of gag p24 and env gp41 were amplified using reverse transcriptase polymerase chain reaction, and genetic subtypes were determined by phylogenetic analysis. RESULTS: Serologic testing confirmed that 63 out of 64 plasma units were positive for HIV 1 group M infection and showed no evidence of HIV-1 group O infections. Genetic subtyping determined that 25 samples were subtype A, three subtype C, 22 subtype D, and nine were heterogeneous for subtypes A and D. CONCLUSIONS: Despite the sequence variation observed in Uganda, commercial EIA based on HIV-1 subtype B proteins detected all the infections. In contrast, a peptide-based assay failed to detect three infections by subtype D viruses. This emphasizes the negative impact of HIV genetic variation on assays that rely on peptides to detect HIV infections. The number of infections with heterogeneous subtype (due to mixed infections or recombinant viruses) is high and reflects the growing complexity of the HIV epidemic in endemic regions where multiple subtypes are present in the population. PMID- 9412703 TI - Clinical implications of autoantibodies in HIV infection. AB - OBJECTIVE: To study the frequency and specificity of autoantibodies in HIV infected subjects and their association with rheumatic manifestations, immunodeficiency, and prognosis. DESIGN: Prospective study of sequentially selected HIV-infected patients. Indirect immunofluorescence reading was performed by two independent observers blinded for the patient diagnosis. Enzyme-linked immunosorbent assay (ELISA) was performed using coded serum samples. SETTING: The study was performed at the Infectious Disease and Rheumatology Divisions of a tertiary care university hospital. PATIENTS: One hundred sequentially selected HIV-infected patients formed group A. Controls included 80 non-HIV-infected high risk individuals (group B), 20 herpesvirus-infected patients (group C), and 30 healthy blood donors (group D). MAIN OUTCOME MEASURES: Patients were followed for 2 years and evaluated for the presence of immunodeficiency, rheumatic manifestations, circulating autoantibodies and total CD4+ cell count. Indirect immunofluorescence was used to investigate antinuclear antibodies, antibodies to native DNA, smooth muscle, parietal cell, glomeruli, thyroid, and neutrophil cytoplasm. Agglutination was used to detect antibodies to erythrocytes and rheumatoid factor. ELISA was used to determine antibodies to cardiolipin and denatured DNA. CD4+ lymphocytes were counted by flow cytometry. Immunoglobulin (Ig) G, IgM and IgA serum levels were determined by radial immunodiffusion. RESULTS: HIV-infected patients presented higher overall frequency of autoantibodies than the other groups. No difference was observed between immunodeficient and asymptomatic HIV-infected patients. The most frequent specificities were antibodies to cardiolipin and to denatured DNA. Ig serum levels did not correlate with the occurrence of autoantibodies. The presence of autoantibodies was associated with lower CD4+ cell counts and with higher mortality within 2 years. Rheumatic manifestations were observed in 35 HIV infected patients and were not associated with the occurrence of autoantibodies or the presence of immunodeficiency. CONCLUSIONS: HIV infection is associated with an increased incidence of autoantibodies. Although not related to the occurrence of rheumatic manifestations, the presence of autoantibodies was significantly associated with lower CD4+ lymphocyte counts and increased mortality, which implies prognostic significance to this phenomenon in the context of HIV infection. PMID- 9412704 TI - Potential risk factors for vertical HIV-1 transmission in Catalonia, Spain: the protective role of cesarean section. The Working Group on HIV-1 Vertical Transmission in Catalonia. AB - OBJECTIVE: To evaluate the roles of certain potential risk factors on the vertical transmission of HIV-1. DESIGN: Prospective registry of infants born to HIV-1-infected women in Catalonia (north-east Spain) from 1987 to 1992. METHODS: A total of 599 infants, born in Catalan hospitals to 520 women who were identified as being HIV-1-infected during gestation or at delivery, were included. Data on mode of delivery, birth weight, gestational age and breast feeding as well as the mother's age, her route of HIV-1 infection, clinical stage and p24 antigenaemia, were recorded. HIV-1 infection status of 489 (82%) of the infants was determined according to the criteria of the US Centers for Disease Control and Prevention. Risk estimates and odds ratio (OR) were calculated and logistic regression was performed. RESULTS: The overall rate of vertical transmission was 18.6% (95% confidence interval, 15.2-22.0%). Multivariate analyses revealed that Cesarean section was associated with a lower rate of vertical transmission (OR = 0.3; P = 0.001), as was maternal HIV-1 infection via injecting drug use (OR = 0.44; P = 0.02). Breast-feeding (OR = 6.9; P = 0.001), very low birth weight (< 1500 g; OR = 6.3; P = 0.001) and p24 antigenaemia (OR = 4.6; P = 0.04) were all related to increased risk. The crude rate of HIV-1 transmission was 6% among Cesarean births compared with 21% for infants born via vaginal deliveries. The population-attributable risk for vaginal deliveries was 61.7%. CONCLUSIONS: The results show a protective effect of Cesarean section in the absence of zidovudine prophylaxis. However, current research should be directed towards the individual and combined effects that antiretroviral agents and Cesarean section may have on mother-to-child HIV-1 infection. PMID- 9412705 TI - Immunological markers in HIV-infected pregnant women. The European Collaborative Study and the Swiss HIV Pregnancy Cohort. AB - OBJECTIVE: To examine immunological markers in HIV-infected pregnant women. DESIGN: Women enrolled in the European Collaborative Study and the Swiss HIV Pregnancy Cohort were followed throughout pregnancy according to similar clinical and immunological protocols. Information was recorded at various times during pregnancy and, in some centres, also 6 weeks and 6 months post-partum. METHOD: Locally-weighted linear regression analysis was used to investigate changes in markers of cellular and humoral immune function during pregnancy and immediately post-partum, taking into account the serial measurement data structure. Women who received zidovudine during pregnancy were excluded. RESULTS: Four hundred and thirty-eight women had two or more measurements during pregnancy or within 6 months of delivery. Twenty-four per cent (106) of the women reported injecting drugs during pregnancy. Mean CD4 and CD8 cell counts declined to a low level 6 months before delivery, increased gradually until delivery and rose sharply to a peak level 3 months post-partum. In contrast, CD4 and CD8 percentages were stable during pregnancy, and increased slightly thereafter. The same pattern was evident for transmitting women, those delivered by Cesarean section, and women who injected drugs during pregnancy, and there was no evidence for an association with immunosuppression. Total immunoglobulin (Ig) G levels declined gradually throughout pregnancy until delivery, and increased in the 6 month post-partum period. Total IgM and IgA levels remained stable throughout pregnancy. CONCLUSIONS: These findings suggest that pregnancy does not accelerate HIV progression, but in view of the intrinsic variability in serial CD4 counts, caution should be exercised when assessing changes in immunological markers in individual pregnant women. PMID- 9412706 TI - Impact of the 1994 expanded World Health Organization AIDS case definition on AIDS surveillance in university hospitals and tuberculosis centers in Cote d'Ivoire. AB - OBJECTIVE: To assess the impact of the 1994 expanded World Health Organization (WHO) AIDS case definition on AIDS surveillance in Cote d'Ivoire. DESIGN: Prospective AIDS case surveillance. METHODS: From March 1994 through December 1996, passive AIDS case surveillance was conducted at the three university hospitals in Abidjan, and active AIDS case surveillance was conducted at the eight tuberculosis (TB) centers in Cote d'Ivoire. Standardized questionnaires were administered and blood samples for HIV serologic testing were collected from the patients evaluated. The numbers of persons who met the modified 1985 WHO clinical AIDS case definition (Bangui definition) and the 1994 expanded WHO AIDS case definition were determined, and the clinical characteristics of these patients were assessed. RESULTS: Of 8648 university hospital patients, 3658 (42.3%) met the clinical and/or the expanded case definition: 744 (20.3%) HIV seropositive persons met only the expanded definition, 44 (1.2%) HIV-seropositive persons met only the clinical definition, 2334 (63.8%) HIV-seropositive persons met both definitions, and 536 (14.7%) HIV-seronegative persons met only the clinical definition. Of 18,661 TB center patients, 9664 (51.8%) met the clinical and/or the expanded definition: 5685 (58.8%) HIV-seropositive persons met only the expanded definition, none of the HIV-seropositive persons met only the clinical definition (by definition), 2625 (27.2%) HIV-seropositive persons met both definitions, and 1354 (14.0%) HIV-seronegative persons met only the clinical definition. CONCLUSIONS: Because of the inclusion of multiple severe HIV-related illnesses into the expanded definition, the number of reportable AIDS cases in HIV-seropositive patients increased 31.3% in the university hospitals, and 217% in the TB centers. The inclusion of HIV seropositivity as a criterion for the expanded definition also enhanced the specificity of AIDS case reporting, eliminating 536 cases in the university hospitals and 1354 cases in the TB centers in HIV-seronegative patients who had clinical signs of AIDS. The use of the 1994 expanded definition for surveillance purposes should be encouraged in areas of the developing world where HIV serologic testing is available. PMID- 9412708 TI - Improving HIV/AIDS disease surveillance in low-income countries. PMID- 9412707 TI - Improved treatment services significantly reduce the prevalence of sexually transmitted diseases in rural Tanzania: results of a randomized controlled trial. AB - OBJECTIVE: To evaluate the impact of improved case management for sexually transmitted diseases (STD) at the primary health care level on the incidence and prevalence of STD. DESIGN: Community-randomized controlled trial. SETTING: Mwanza region, Tanzania. SUBJECTS: A random cohort of about 1000 adults aged 15-54 years from each of 12 communities, in six matched pairs. One member of each pair was assigned at random to receive the intervention, and the others served as a comparison community. This cohort was surveyed at baseline and at follow-up 2 years later. About 100 antenatal clinic attenders were also studied in each community on two occasions: the first shortly after the implementation of the intervention, and the second approximately 1 year later. INTERVENTION: Improved services were established for the management of STD, using the syndromic approach, in rural health units. RESULTS: A total of 12,534 individuals were enrolled in the cohort study, of whom 8844 (71%) were seen again 2 years later. The prevalence of serological syphilis (rapid plasma reagin titre > or = 1:8, Treponema pallidum haemagglutinin assay positive) was 6.2% in both intervention and comparison communities at baseline. At follow-up it was 5.0% in the intervention community and 7.0% in the comparison community [adjusted relative risk (RR), 0.71; 95% confidence interval (CI), 0.54-0.93; P < 0.02]. The prevalence of urethritis in males did not differ significantly between intervention and comparison groups at follow-up, but the prevalence of symptomatic urethritis was reduced by about 50% (adjusted RR, 0.51; 95% CI, 0.24 1.10; P = 0.08). There was no significant difference between the groups in the incidence of self-reported STD symptoms over the last year of the follow-up period, or in the prevalence of any STD in antenatal clinic attenders. CONCLUSION: The reduction in HIV incidence previously reported in this intervention study can be attributed to a reduction in the duration, and hence the prevalence of symptomatic STD. PMID- 9412709 TI - Identification of a novel HLA-A24-restricted cytotoxic T-lymphocyte epitope within HIV-1 Nef. PMID- 9412710 TI - Potential underestimation of HLA-C-restricted cytotoxic T-lymphocyte responses. PMID- 9412711 TI - Changes in CXC-chemokine receptor-4 expression during HIV-1 replication are independent of CD4 modulations. PMID- 9412712 TI - A multicentre prospective study for the polymerase chain reaction detection of Toxoplasma gondii DNA in blood samples from 186 AIDS patients with suspected toxoplasmic encephalitis. Bio-Toxo Study Group. PMID- 9412713 TI - Antiretroviral therapy and HIV-related retinal microangiopathy. PMID- 9412714 TI - Cutaneous vasculitis due to human parvovirus B19 in an HIV-infected patient: report of a case. PMID- 9412715 TI - Kaposi's sarcoma patients in the Swiss HIV Cohort Study with and without detectable human herpesvirus 8 in peripheral blood mononuclear cells. PMID- 9412716 TI - Mucosal Kaposi's sarcoma following protease inhibitor therapy in an HIV-infected patient. PMID- 9412717 TI - Safety and efficacy of two different triple drug combinations in which either lamivudine or didanosine were administered with stavudine plus indinavir. PMID- 9412718 TI - HIV-1 subtype C epidemic in Ethiopia. PMID- 9412719 TI - HIV-1 subtypes: implications for epidemiology, pathogenicity, vaccines and diagnostics. Workshop Report from the European Commission (DG XII, INCO-DC) and the Joint United Nations Programme on HIV/AIDS. AB - Forty-three AIDS scientists from Europe, Africa, the United States, Canada, India and China met in Dar es Salaam, Tanzania to discuss the implications of the global variation of HIV (list of participants included in Appendix). This meeting followed an earlier meeting of the Joint United Nations Programme on HIV/AIDS, held in 1996 in Berlin, Germany [1], in which HIV genetic variability was considered in relation to transmissibility. During the Tanzania meeting, available data pertaining to the biological consequences of HIV genetic variation and its ramifications with regard to epidemiology, diagnostics, classification, and vaccine design were systematically reviewed. There was consensus that classification based on genetically defined subtypes provides an important framework for making advances on understanding viral biology and immunology, and for vaccine development. In addition, other groupings of viruses based on immunological and biological characteristics would be also valuable and help to further refine our understanding of the implications of variability. Key elements of the discussion are summarized here in the context of a review of the current literature. PMID- 9412720 TI - Characterization of antibodies generated against a conserved portion of oviductal glycoprotein (OGP) and endogenous hamster OGP and their ability to decrease sperm binding to the zona pellucida in vitro. AB - PROBLEM: The effect of antibodies generated against hamster oviductal glycoprotein (OGP) on sperm binding to the zona pellucida (ZP) was evaluated. METHOD OF STUDY: Antibodies against a 17-amino-acid sequence of the OGP core protein (amino acids 52-68) and the denatured hamster OGP protein were generated, characterized, and tested in an in vitro sperm binding assay. RESULTS: Sperm binding was significantly decreased (P < 0.05) when oviductal oocytes were incubated for 2 hr with 4 or 8 mg/ml of immune IgG of both antibodies when compared with normal rabbit IgG. A fluorescence assay showed binding of both antibodies to the endogenous OGP associated with the ZP of ovulated hamster oocytes. CONCLUSIONS: These results suggest that OGP may be a potential immunocontraceptive target because both antibodies significantly decreased sperm binding to the ZP of oviductal oocytes. Immunocontraception may be accomplished by attempting to generate active immunity to a recombinant OGP, to the region selected in this study (amino acids 52-68) or to some other region of the core protein. PMID- 9412721 TI - The presence of a C1-inhibitor-like molecule (C1-INH-L) on human sperm: its involvement in sperm motility. AB - PROBLEM: An 88-92-kDa C1-inhibitor-like molecule (C1-INH-L) was previously identified to elicit cytotoxic sperm antibody response in infertile men and women. Here, we document that it is present on the human sperm surface and could be detected by an enzyme-labeled immunoglobulin G (IgG) fraction of anti-human C1 INH antibody. METHOD OF STUDY: Western blot analysis, enzyme-lined immunoadsorbent assay (ELISA) and computerized sperm motion analysis. RESULTS: The existence of C1-INH-L on the sperm surface is calcium independent. Phosphatidylinositol-specific phospholipase C (PIPLC), EDTA, and acid (pH 3.0) could not remove the C1-INH-L from sperm, but trypsin did. Activated C1s was able to bind to the sperm surface. Immunofluorescence studies localized the protein to the head and midpiece of the sperm membrane. The C1-INH-L exists on both uncapacitated and capacitated sperm surfaces, which suggests that this protein is a sperm-surface protein. The heat-treated (56 degrees C, 30 min) IgG fraction of anti-C1-INH greatly reduced the percentage of motile spermatozoa and the progressive and path velocities in the absence of complement. CONCLUSION: Our data suggest that C1-INH is a sperm membrane-anchored protein that may have complement and sperm motility regulatory function. PMID- 9412722 TI - A tissue type transglutaminase in human seminal plasma. AB - PROBLEM: Initial studies from this laboratory of human seminal plasma (SePl) permitted the presumptive identification of the participation of transglutaminase (TGase) and prostaglandins as principal molecules contributing to the immunoregulatory activity of SePl. As a step toward confirmation and extension of these studies. SePl TGase has been partially purified, characterized, and localized. METHOD OF STUDY: An enzyme-enriched and partially purified preparation of SePl TGase obtained by molecular sieve and ion-exchange chromatography and the polyclonal antibody to this preparation were characterized by enzymatic analysis and evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE); immunoprecipitation and immunofluorescence (IF). RESULTS: A Ca(2+) dependent, thrombin-independent, TGase enzyme from SePl was identified. A pH of 7.5 and a concentration of 2 mM calcium chloride were determined to be optimal for ascertaining the SePl TGase activity in a filter paper assay. Immunoprecipitation using polyclonal antibody prepared with a semipurified TGase preparation, concurrently comparing increasing serum dilution and enzyme activity, revealed a predominant protein band on SDS-PAGE of 83 kDa. IF studies using the polyclonal antibody on human prostate tissue showed reactivity with a variety of epithelial and stromal components. A significant (P < 0.05) correlation between the biochemical, i.e., enzyme activity of the SePl TGase active preparation, and immunologic activity, i.e., indirect IF staining titre of antibody to acinar epithelial cells and luminal contents, was observed. CONCLUSIONS: The results confirm and extend initial studies of the presumptive identification of a tissue type TGase associated with the immunoregulatory activity of human SePl and permit the characterization and immunohistologic localization of this macromolecule to the prostate. These observations provide a basis for further investigation of the immunoregulatory role of SePl and prostate TGase in the biology of reproduction and associated pathoses. PMID- 9412723 TI - Peritoneal cellular immunity and endometriosis. AB - PROBLEM: An immunologic basis has long been considered to be very important in the pathogenesis of endometriosis. Interactions of the peritoneal cells, which comprise macrophages, B cells, T cells, natural killer (NK) cells, and retrograde endometrial cells, are critical, but remain controversial, for exploring the pathogenesis of endometriosis. METHOD OF STUDY: Accumulated data from the literature were reviewed, and our data were analyzed. RESULTS: The data show that peritoneal macrophages are activated by the recurrent reflux of menstrual shedding. Humoral and local endometrial autoantibodies are detected in patients with endometriosis, but B cells are not quantitatively increased. There is decreased NK cell activity in the peritoneal cavity and peripheral blood, and this decreased activity may be related to the failure to clear out the ectopic endometrial tissue. Peritoneal T cells are predominant by Th1 inflammatory cells, and these cells are impaired because of a decrease in activation (especially HLA DR+CD4+CD3+ population) and in the production of interleukin-2. Inflammatory cytokines such as interleukin-1, interleukin-6, and tumor necrosis factor-alpha are elevated in the peritoneal fluid of women with endometriosis. CONCLUSIONS: The peritoneal NK and T lymphocytes are suppressed in women with endometriosis, but whether these immunologic deviations are the cause or the result of endometriosis is still unclear. Further studies are required to determine what role immunologic factors play in the pathophysiology of endometriosis. PMID- 9412724 TI - Transferrin receptor (CD71) expression in peritoneal macrophages from fertile and infertile women with and without endometriosis. AB - PROBLEM: Hyperactivated macrophages are implicated in the pathophysiology of endometriosis-associated infertility. This study investigates transferrin receptor expression (CD71) as a marker of hyperactivity in peritoneal macrophages of infertile patients with minimal to mild endometriosis (group 1, n = 25). METHOD OF STUDY: Expression of the activation antigen CD71 on peritoneal fluid macrophages was determined by a specific monoclonal anti-CD71 antibody using indirect immunofluorescence technique and was analyzed by flow cytometry. Three different control groups of women were used: women with unexplained infertility (group 2, n = 25), fertile women with endometriosis (group 3, n = 10), and fertile women without endometriosis (group 4, n = 25). RESULTS: The percentage of CD71 positive cells was significantly increased in infertile women with endometriosis as compared with the three control groups. There were no differences among groups 2, 3, and 4 with respect to the percentage of CD71 positive macrophages. CONCLUSIONS: Our results favor the concept that hyperactivated macrophages play a role in the pathophysiology of endometriosis associated subfertility, a feature which is lacking in patients with unexplained infertility. PMID- 9412725 TI - Cytokine production by lymphocytes in pregnancy. AB - PROBLEM: In the presence of progesterone lymphocytes of pregnant women release a 34-kDa protein named the progesterone-induced blocking factor (PIBF). PIBF mediates the immunomodulatory and anti-abortive effects of progesterone and its presence is related to the outcome of pregnancy. PIBF induces production of Th2 type cytokines by activated lymphocytes. The in vivo relationship between PIBF- and cytokine production of pregnancy lymphocytes and the outcome of pregnancy was investigated. METHOD OF STUDY: Interleukin (IL)-12 and IL-10 production and PIBF expression in peripheral lymphocytes of 111 healthy pregnant women and 120 women at risk for premature pregnancy termination were detected by immunocytochemistry. RESULTS: We found increased IL-12 and low PIBF and IL-10 expression on lymphocytes of "risk" patients, and a high rate of IL-10 and PIBF positivity on lymphocytes from healthy pregnant women. The cytokine production pattern of the lymphocytes was related to the presence or absence of previous abortions as well as to the outcome of pregnancy. CONCLUSION: These data suggest the involvement of an altered cytokine production pattern in the immunologic effects of progesterone. PMID- 9412726 TI - Phenotypic analysis of adhesion molecules in first-trimester decidual tissue from chorion villus samples. AB - PROBLEM: First-trimester decidua contain CD56bright/CD16neg lymphocytes that phenotypically resemble a subset of peripheral blood natural killer (NK) cells. Factor controlling their localization to decidua are unknown, but they may relate to trophoblast invasion, endometrial decidualization, and adhesion molecule expression. METHOD OF STUDY: Ninety-one chorion villous samples (CVS) were screened for the presence of decidua, and selected specimens were analyzed for the expression of adhesion molecule pairs using a panel of monoclonal antibodies. RESULTS: Lymphoid cells in CVS-derived decidua expressed CD56, PECAM, intracellular adhesion molecule-1 (ICAM-1), and the integrins LFA-1, alpha E beta 7, and alpha 4 beta 1, and co-receptors for these molecules were expressed by decidual stromal cells, invasive trophoblasts, and venule endothelium. Some endothelium expressed a cuboidal phenotype. CONCLUSIONS: The expression of complimentary pairs of adhesion molecules by maternal lymphoid cells and by either extravillous cytotrophoblast or decidual stromal cells supports the hypothesis that these cells interact within decidua. Also, both LFA-1 and alpha 4 beta 1 may contribute to decidual localization of CD56bright NK cells. PMID- 9412727 TI - Lymphokine-activated killer (LAK)-mediated lysis of sequentially isolated ovarian cancer cell lines. AB - PROBLEM: Understanding immunologic eradication of cancer is hampered by failure to explore tumor evolution. Cell surface molecules may alter with therapy and disease progression. These alterations can translate into variable susceptibility to immune-mediated cell lysis. METHOD OF STUDY: Cell lines from a patient with ovarian carcinoma isolated at surgical debulking (UL-3A), during chemotherapy (UL 3B), and after progression (UL-3C) were used to study changes in cell lysis by natural killer (NK) and lymphokine-activated killer (LAK) cells. The role of adhesion molecules ICAM-1, LFA-3, and glycoproteins in the demonstrated differential killing was also examined. RESULTS: An inverse relationship between attachment and lysis was demonstrated. UL-3C, the most sensitive to lysis (50%), attached the least lymphocytes (40%), whereas UL-3A, the least sensitive (33%), attached the most lymphocytes (71%). A correlation with ICAM-1 and LFA-3 expression was not demonstrated. CONCLUSION: Ovarian cancer cells evolve throughout the disease course, and this may manifest as differential sensitivity to immune-mediated cell lysis. PMID- 9412728 TI - Effect of peptide to carrier ratio on the immune and ovarian response to inhibin immunization in cattle. AB - We report on the effects of the peptide to carrier ratio on the immune and biological response to inhibin immunization in cattle. A peptide sequence from the alpha C-subunit of bovine inhibin was synthesized and conjugated to human serum albumin (HSA) at ratios of 4.3 moles (L) and 13.1 moles (M) of peptide per mole of HSA. Hereford-cross heifers (n = 6 per group) were injected with 3 mg of one of the peptide conjugates at primary, followed by a booster injection (1.5 mg) 11 weeks later. Control heifers (n = 6) were injected with HSA only. Blood samples were taken at regular intervals to measure antibody titre. Ovulation rate was measured by ultrasonography. Antibodies were generated in both peptide immunized groups. Control heifers and group L heifers had 1 ovulation at all ovulatory cycles monitored. Ovulation rate was increased (P < 0.05) in group M immunized heifers, with four of six heifers having twin ovulations in the first cycle following boost. These data support those of previous studies which indicated that immunization against the alpha C-subunit of bovine inhibin significantly disrupted the mechanism(s) controlling ovulation rate in cattle. It also indicates that both the immune and associated biological response is dependent on the nature of the conjugate used for immunization, specifically the ratio of peptide to carrier. PMID- 9412729 TI - An evaluation of transovarian uptake of metabolites using arterio-venous difference methods in dairy cattle. AB - Arterio-venous (A-V) difference techniques were used in cattle to examine ovarian energy metabolism, cholesterol uptake and steroid hormone outputs. Catheters were inserted into the ovarian vein and facial artery, and Transonic flow transducers were placed around the ovarian A-V plexus. Further, in some cows, the effects of a challenge with GnRH were examined. Glucose uptake and lactate output were significant in most individual cows. Nonesterified fatty acids (NEFA) uptake were not significant in any cow in dioestrus. Ovarian uptake of beta-Hydroxy-butyrate (3-OHB) was significant in 4 cows in dioestrus. Cholesterol uptake was significant in only 1 cow. Oxygen uptake was significant in all cows at all stages of the oestrous cycle. All cows had significant output of progesterone and oestradiol-17 beta. These data show that the bovine ovary utilises significant amounts of glucose, and Respiratory quotient (RQ) estimates demonstrated that glucose was the primary fuel used by the ovary. The significant output of lactate suggested that anaerobic pathways were mainly used for glucose oxidation. The observed uptakes of 3-OHB indicated that the ovary utilises 3-OHB as a source of energy. Cholesterol uptake was not a rate-limiting factor for steroid hormone production in the ovary. Despite the high metabolic rate in the luteal ovary, the small difference in PO2 between arterial and ovarian venous blood indicated that the ovary consumes only a small proportion of available oxygen. GnRH had no significant effect on the uptake of metabolites and energy metabolism, but it increased OBF and the output of progesterone and oestradiol-17 beta. The use of A V methods to determine the metabolic needs of the ovary is useful in understanding the means by which nutrition can influence fertility. PMID- 9412730 TI - Usefulness of polyethylene glycol for cryopreservation by vitrification of in vitro-derived bovine blastocysts. AB - A series of five experiments measured the high survival of bovine blastocysts produced in vitro after cryopreservation by vitrification. The vitrification solution (designated VS) contained 40% (v/v) ethylene glycol, 6% (w/v) polyethylene glycol and 0.5 M sucrose in phosphate-buffered saline. Embryos developed in vitro at Days 7 and 8 (Day 0 = insemination day) were exposed in one step to VS for 1 min or two steps with 10% ethylene glycol for 5 min and then VS for 1 min. In both cases, the embryos were finally cryopreserved in liquid nitrogen. After the embryos were warmed rapidly and the VS solution diluted, the survival rates were assessed by monitoring hatching rate in vitro. They were 13.0% for the one-step and 72.7% for the two-step procedures (P < 0.001). When embryos were exposed to individual solutions containing 6% (w/v) of each of 4 macromolecules (polyethylene glycol, BSA, polyvinylpyrrolidone or Ficoll) in the two-step protocol and then cryopreserved, the survival rates were 79.3, 34.8, 41.4 and 57.1%, respectively. After embryos had been exposed to the VS in two steps and then cryopreserved, there were no significant differences in survival rates when the solutions were diluted with or without sucrose. These results indicated that a vitrification solution containing polyethylene glycol can be used for cryopreservation of bovine blastocysts produced in vitro, and that a two step addition of VS improved the in vitro survival of post-warming embryos. It was also shown to be possible to dilute post-warming embryos directly without the use of sucrose solution. PMID- 9412731 TI - The effects of bovine serum albumin and fetal bovine serum on the development of pre- and postcleavage-stage bovine embryos cultured in modified CR2 and M199 media. AB - The effects of protein supplementation on bovine embryo development in vitro was evaluated using a 4 x 2 factorial arrangement with ten replications. A total of 6438 oocytes collected from abattoir ovaries were used. Bovine serum albumin (BSA) and fetal bovine serum (FBS) were added in various combinations to simple (modified CR2) and complex (M199) media during culture of precleavage-stage IVM/IVF-derived ova from 18 h after insemination to 72 h and postcleavage-stage embryos after 72 h of culture. Cleavage rates did not differ (p > 0.05) between media supplemented with FBS or with BSA. However, the postcleavage development to the blastocyst stage of in vitro-derived bovine embryos is better in media supplemented with FBS than BSA. A greater (p < 0.05) proportion of cleaved oocytes developed to blastocysts and hatched blastocysts in media supplemented with FBS during postcleavage culture. The percentage of embryos that stopped development at the morula stage was significantly (p < 0.05) greater in media supplemented with BSA during postcleavage culture. Viability of blastocysts produced in CR2 and M199 supplemented with FBS were further assessed by transfer to recipients. In CR2, 25 transferred blastocysts resulted in seven pregnancies and the birth of three normal calves. In M199, 24 transferred blastocysts resulted in five pregnancies and the birth of two normal calves. There was no difference (p > 0.05) in rate of embryo development between CR2 and M199. PMID- 9412732 TI - The use of oxytocin for the reduction of cow placental retention, and subsequent endometritis. AB - Oxytocin was administered to reduce incidence of retained placenta and uterine infections that could delay subsequent conception. Three hundred and fifty multiparous Friesian cows, each with spontaneous delivery of a single calf were divided randomly into two groups. Some (n = 175) were injected with 30 IU of oxytocin immediately after delivery and again 2-4 h later, while the remainder formed an untreated control group. The placental retention 24 h after parturition was 24.6% and 10.9% in control and treated animals respectively (P < 0.01). Endometritis occurred in 51.6% of the animals following placental retention as compared to 10.4% of those with normal expulsion of the fetal membranes (P < 0.001). A comparison of reproductive indices showed a statistically significant improvement of fertility in treated cows with the average interval from calving to conception being reduced from 124.4 d to 93.7 d (P < 0.0001). PMID- 9412733 TI - Effect of adrenocorticotrophic hormone on gonadotrophin releasing hormone-induced luteinizing hormone secretion in vitro. AB - An in vitro perifusion study investigated the effect of different forms of adrenocorticotrophic hormone (ACTH) on gonadotrophin releasing hormone (GnRH) induced luteinizing hormone (LH) secretion, particularly GnRH self-priming, and oestradiol sensitisation of the ovine pituitary. Fragments of pituitaries were obtained from mixed-breed adult nonpregnant female sheep (without corpora lutea, unless otherwise stated). The amount of LH released by different doses of GnRH (2.5 x 10(-10) M (n = 9 chambers), 1 x 10(-10) M (n = 9), or 5 x 10(-11) M (n = 6)) was evaluated by giving two GnRH pulses (5 min each) 2 h apart. In a duplicate set of chambers, ACTH1-24 (5 x 10(-7) M) was included in the perifusate 0.5 h before the first GnRH challenge. Potassium chloride (KCl; 100 mM) was administered 2 h after the second GnRH challenge to assess the viability of the tissue and the size of the releasable LH pool. Results were expressed as percentage of LH secretion. The influence of ACTH1-24 on oestradiol sensitisation was also examined using pituitaries obtained during the luteal phase. Pituitary tissues were perifused throughout with 1 x 10(-9) M or 6 x 10(-11) M oestradiol in the medium. The LH response to the second GnRH challenge (GnRH 2) was significantly greater (p < 0.01) than after the first (GnRH 1) at the highest dose of GnRH (2.5 x 10(-10) M; 2547 +/- 804 vs. 4547 +/- 1013%), but at the lower doses (1 x 10(-10) M or 5 x 10(-11) M), the self-priming effect of GnRH was not evident (3016 +/- 550 vs. 2932 +/- 490% and 841 +/- 205 vs. 711 +/- 87%). Treatment with ACTH1-24 (5 x 10(-7) M) did not affect tonic LH secretion nor the LH response to the first or second GnRH challenge at any of the GnRH doses tested. The LH released in response to KCl was also similar from control and ACTH1-24-treated tissue at all GnRH doses. Both lower doses of GnRH (1 x 10(-10) M or 5 x 10(-11) M) produced the self-priming effect when the pituitary tissue was sensitised with the higher dose of oestradiol (1 x 10(-9) M; 1711 +/- 239 vs. 5085 +/- 1307%, and 1502 +/- 376 vs. 2619 +/- 629%). In the presence of lower concentrations of oestradiol (6 x 10(-11) M), self-priming was observed only after the higher dose of GnRH (1 x 10(-10) M; 1293 +/- 214 vs. 2865 +/- 436%), not the lower dose (5 x 10(-11) M; 985 +/- 203 vs. 1271 +/- 436%). In spite of these differences, ACTH1-24 treatment did not affect LH secretion (neither basal nor potassium-induced). The effect of ACTH1-39 (1 x 10(-8) M or 5 x 10(-7) M; n = 6 chambers per combination) on GnRH-induced LH secretion was examined using the higher (2.5 x 10(-10) M) or lower dose of GnRH (1 x 10(-10) M), with or without oestradiol sensitisation (1 x 10(-9) M). At the lower dose (1 x 10(-8) M), ACTH1 39 influenced neither tonic nor GnRH-induced LH secretion. The LH released by KCl was also similar to the control and ACTH-treated tissue. In contrast, the higher dose of ACTH1-39 (5 x 10(-7) M) increased tonic LH secretion immediately after inclusion in the medium (104 +/- 3 vs. 161 +/- 20%), but suppressed the GnRH self priming effect after 2.5 x 10(-10) M, i.e., the LH responses to GnRH 1 and 2 were similar (1786 +/- 294 vs. 1553 +/- 373%). However, the LH response to KCl was not significantly different (p > 0.05) between the control and ACTH-treated tissues (2333 +/- 286 vs. 2638 +/- 431%). When the effect of this higher dose of ACTH1-39 on oestradiol-priming was investigated, ACTH increased tonic LH secretion but suppressed the self-priming effect of GnRH (1 x 10(-10) M GnRH; 945 +/- 274 vs. 922 +/- 323%; p > 0.05), and decreased (p < 0.05) the LH released in response to KCl compared to the controls (1803 +/- 409 vs. 4302 +/- 1017%). In summary, in vitro, ACTH1-24 did not affect either tonic LH secretion, the GnRH self-priming effect, or oestradiol sensitisation. The entire ACTH1-39 increased tonic LH secretion, but reduced GnRH self-priming and oestradiol sensitisation. (ABSTRACT TRUNCATED) PMID- 9412735 TI - Quantitative aspects of sperm:egg interaction in chickens and turkeys. AB - Spermatozoa embedded in the outer perivitelline layer and points of hydrolysis (holes) produced by spermatozoa in the inner perivitelline layer of chicken and turkey eggs were found to be evenly distributed and linearly correlated (r = 0.80 for both species) throughout the layers from most regions of the egg, except from those directly over the germinal disc, in which there were more holes. In turkey eggs there appeared to be relatively fewer perivitelline spermatozoa, since many had degenerated beyond recognition. In eggs from both species, there were approximately 25 times more holes mm-2 in the inner perivitelline layer from over the germinal disc region than that from other regions of the egg. The relationship between these two frequencies could also be described as linear (r = 0.81 for chicken and 0.78 for turkey eggs), although there was some evidence for a saturation effect for holes over the germinal disc. The fertile status of eggs was shown to be a function of all of the above parameters. Eggs from both species had a 50% probability of being fertile when around 3 spermatozoa penetrated the inner perivitelline layer over the germinal disc and showed maximum fertility when more than 6 spermatozoa penetrated this region. Spermatozoa in the outer perivitelline layer and holes in the inner perivitelline layer from regions other than over the germinal disc could also be used to predict fertility, although with less certainty. Since the number of spermatozoa interacting with the egg reflects the numbers of those stored in the uterovaginal sperm storage tubules, the relationships derived in this work should be useful for understanding how fertility in chickens and turkeys is a function of oviducal sperm storage and transport. PMID- 9412734 TI - Factors affecting electrical activation of porcine oocyte matured in vitro. AB - This work was undertaken to improve conditions for in vitro maturation and activation of porcine oocytes. Experiments were designed to compare: (i) electrical pulse frequency, (ii) methods of oocyte preparation, (iii) maturation conditions, and (iv) electrical poration medium on development. Oocytes were harvested by follicle dissection or aspiration, co-cultured with follicle shells in M199 based medium with or without media changes at 38.5 degrees C in 5% CO2 under non-static conditions for 48 h and electroactivated using single or multiple pulses (current strength 1.0 kV/cm for 50 microseconds in 0.28 M inositol or mannitol based media with 10 mM histidine) at different time intervals. The results showed: (i) neither the pulse frequency nor the pulse interval influenced rates of pronuclear formation but multiple pulse activation (3 pulses at 5 min intervals) induced a higher incidence of development and progression through the 4-cell block in contrast to one pulse activation; (ii) both the rate of nuclear maturation (88.6% vs. 77.6%) and post-activation cleavage (89.8% vs. 67.4%) were higher (P < 0.05) when oocytes were collected by follicle dissection rather than by aspiration; (iii) while changing to a hormone free medium at 24 h was without effect on maturation (91.9% vs. 91.7%), rate of cleavage (81.6% vs. 72.3%, P < 0.05) at 24 h was enhanced by the medium change; and (iv) oocytes activated with 3 pulses 5 min apart in mannitol based medium at 48-49 h and at 53-54 h formed pronuclei at a comparable rate but subsequent parthenogenetic development was higher in the older eggs. By contrast, inositol based medium supported development of young and old eggs equally well. Calcium and magnesium ions are, however, necessary in both mannitol and inositol media for activation of porcine oocytes matured in vitro. The present results suggest that optimal parthenogenetic activation and early development of IVM pig oocytes could be obtained if oocytes are harvested by dissection, cultured for 24 h in hormone-containing medium before being placed in hormone free medium and activated at 48 h in inositol based medium using a three pulse activation system. PMID- 9412736 TI - Intraepithelial gamma delta T cells are closely associated with apoptotic enterocytes in the bovine intestine. AB - The T cell population in the intestinal epithelium, comparable in size to the T cell pool in the spleen, is characterized by the predominant distribution of T cells bearing gamma delta T cell receptors. To determine the functional significance of the intraepithelial lymphocytes, we observed gamma delta T cells present in the jejunal epithelium in cattle, in which there is predominance of gamma delta T cells. Immunohistochemistry of frozen sections demonstrated that gamma delta T lymphocytes were densely distributed in the villous epithelium but there were fewer in the lamina propria and they were not present in the crypt epithelium. Ultrastructurally, intraepithelial gamma delta T cells were characterized by possessing electron-dense granules and interdigitating with enterocyte cytoplasm. Enterocytes, which were inserted by processes of intraepithelial lymphocytes or contacted by their cell bodies, showed morphologic changes seen in apoptotic cell death, such as elevated electron density of the cytoplasm and condensation of the chromatin. Apoptotic cells and cell debris were found in macrophages, which gathered in the subepithelial region of villus tips. These findings suggest that in the small intestine of cattle, gamma delta T cells are involved in the renewal of epithelial cells by inducing apoptosis of epithelial cells. PMID- 9412737 TI - Destruction of olfactory inputs affects the morphogenesis of the telencephalon in rats. AB - We unilaterally destroyed the nasal radix of rat embryos on day 15.5 of gestation (E15.5) in utero so as to block the olfactory inputs to the ipsilateral forebrain vesicle. The embryonic brains were examined after 6 days' survival (E21.5). In the deafferented half of the brain, LHRH neurons were significantly reduced in number, indicating the successful blocking of the olfactory input. On the deafferented side, the olfactory bulb failed to develop, and the telencephalic hemisphere, small in size, accompanied various histogenetic retardations in the primary olfactory cortex, in the cortical plate, and in the hippocampal formation. The striatum revealed remarkable structural differences between the ipsilateral and contralateral sides: on the ipsilateral side, the striatum was small in size and displayed numerical reductions of immunoreactive tyrosine hydroxylase (TH) fibers and substance P (SP) neurons in comparison with those in the contralateral one; in the substantia nigra, TH neurons and SP fibers were less numerous on the deafferented side. There were no remarkable differences in the distribution of TH neurons in the hypothalamus. In view of these sequential histogenetic alterations, it can be assumed that the olfactory inputs play a key role in the telencephalic morphogenesis. PMID- 9412738 TI - Perineuronal sulfated proteoglycans and cell surface glycoproteins in adult and newborn mouse brains, with special reference to their postnatal developments. AB - Sections of the retrosplenial cortex from adult and newborn mouse brains were observed with a light microscope. The retrosplenial cortex of the adult animals contained many neurons (10% of the total), including some dark neurons, with perineuronal sulfated proteoglycans detectable with cationic iron colloid and aldehyde fuchsin. The retrosplenial cortex of the adult animals also contained many neurons (10% of the total) with cell surface glycoproteins reactive to lectin Vicia villosa, soybean or Wisteria floribunda agglutinin. Double staining showed that the majority (75%) of the neurons labeled with lectins were stained with cationic iron colloid, and that some (25%) of them were not stained with this colloid. Double staining also showed that some (25%) of the neurons stained with cationic iron colloid were not labeled with lectins. These findings indicate that the perineuronal sulfated proteoglycans are, at least partly, independent from the cell surface glycoproteins. Observations of the sections from the newborn animals revealed that the perineuronal sulfated proteoglycans were produced by the associated satellite astrocytes 3-4 weeks after birth, and that the cell surface glycoproteins were produced by the associated nerve cells at earlier stages, or 2-3 weeks after birth. Dark neurons began to appear 3-4 weeks after birth. These dark neurons or their Golgi complexes were also reactive to lectins, suggesting the production of cell surface glycoproteins. PMID- 9412739 TI - Heterogeneity of pituitary gonadotrope cells in male rats. AB - Mammalian gonadotrope (Gn-) cells producing FSH and LH show great variability in their cytological characteristics. To gain a closer view of the corresponding phenotypes, Gn-cells of male rats were investigated by various methods including immunocytochemistry on serial semithin sections, quantitative immunocytochemistry, three-dimensional reconstructions, sequential semithin-/thin section technique, routine electron microscopy, and immuno-electron microscopy. Gn-cells were immunostained with FSH and LH antisera and with antibodies raised against synthetic N- and C-terminal sequences of chromogranin A (CgA) and secretogranin II (SgII), and four phenotypes of Gn-cells could be identified. The great majority of Gn-cells were densely immunoreactive for both gonadotropins and both chromogranins and accordingly classified as "typical" gonadotropes; a second type was less immunoreactive for LH and therefore was termed "LH-poor". The two remaining phenotypes showed ultrastructures suggesting an increased synthetic activity. One of them was characterized as a "FSH/CgA-poor" gonadotrope. The other, also poor in CgA, morphologically corresponded to "signet-ring" cells previously known only in gonadectomized animals. The two types of secretory granules present in male rat Gn-cells showed a strict segregation of chromogranin immunoreactivities: CgA was confined to the large, less opaque granules, whereas SgII, to the small, electron-dense granules. Consequently the two "CgA-poor" Gn cell subtypes exhibited only a very few large-sized secretory granules. A correlation was suggested between high cellular activity and a decrease in CgA in Gn-cells. The reasons for the morphological and functional heterogeneity of Gn cells remain enigmatic. Presumably, they either differ in the susceptibility for GnRH and testosterone or are differently regulated by paracrine factors. PMID- 9412740 TI - Atomic force microscopic studies of isolated collagen fibrils of the bovine cornea and sclera. AB - Isolated collagen fibrils from the bovine cornea and sclera were investigated by atomic force microscopy (AFM) in a non-contact mode. AFM imaging visualized the surface topography of both corneal and scleral collagen fibrils with quantitative information on their height and width. The corneal collagen fibrils had a height of 15.6 +/- 1.5 nm and a D-periodicity of 63.9 +/- 0.5 nm. On the other hand, the height and D-periodicity of scleral collagen fibrils were 74.2 +/- 55.7 nm and 65.4 +/- 0.7 nm, respectively. A periodic banding pattern of grooves and ridges was found in individual fibrils, although the groove depth was 2.81 +/- 0.29 nm in the cornea and 5.47 +/- 0.66 nm in the sclera. When collagen fibrils were treated with acetic acid, they swelled and untwisted into subfibrils. The AFM is useful to analyze surface morphology of collagen fibrils and their subfibrils at high resolution with quantitative information. PMID- 9412741 TI - Polymorphism of Merkel cells in the rodent palatine mucosa: immunohistochemical and ultrastructural studies. AB - Our pervious electron microscopic studies indicated that Merkel cells (MCs) in the gerbil palatine mucosa were polymorphic, possibly reflecting different function. In order to verify and extend this evidence, the shape of and the innervation to MCs in the palatine mucosa of six different species of rodents including the Mongolian gerbil and the rat were examined by immunohistochemistry and transmission electron microscopy. Immunohistochemistry using anti-cytokeratin 20 (CK20) antibody revealed that in the gerbil palatine mucosa, approximately half of MCs were dendritic. Confocal laser scanning microscopy after double labeling with anti-cytokeratin and anti-PGP 9.5 or anti-Na+/K(+)-ATPase beta 1 subunit antibodies indicated that most of the dendritic MCs (DMCs) in these mucosae were free of innervation. Electron microscopy showed that all species of rodents examined contained abundant dendritic MCs as well as roundish (oval to round) MCs (RMCs) with typical innervation. Secretory granules of the RMCs were usually concentrated at the synaptic site, whereas those of the DMCs tended to accumulate in the tips of the cytoplasmic processes and in the cytoplasm facing the basal lamina. Some MCs showed features intermediate between those of the RMC and DMC. These results indicate that MCs in rodent palatine mucosae are consistently polymorphic, and that DMCs may represent a distinctive subset with specific, presumably including endocrine and paracrine, functions different from those of RMCs. PMID- 9412742 TI - Neuronal circuitry between the inferior mesenteric ganglion and lower intestine of the dog. AB - Neuronal circuitry between the inferior mesenteric ganglion (IMG) and the distal colon as well as the rectum, forming the intestino-intestinal reflex pathway, was investigated in the dog using immunohistochemistry combined with retrograde tract tracing and denervation experiments. Virtually all IMG neurons were tyrosine hydroxylase (TH)-immunoreactive. Of these ganglionic neurons, about 64% were also immunoreactive for calbindin (Calb), some 35% for neuropeptide Y (NPY), and 2% for vasoactive intestinal peptide (VIP). The retrograde tracer experiments revealed that both Calb/TH neurons and NPY/TH neurons projected to the distal colon and the rectum. In these intestinal walls, Calb/TH positive varicose fibers were found in the myenteric and submucous ganglia as well as in the longitudinal muscle layer, while NPY/TH positive fibers were mainly distributed around the vascular walls. Around Calb/TH neurons of the IMG, abundant varicose nerve fibers immunoreactive for VIP, dynorphin (DYN), calcitonin gene-related peptide (CGRP), enkephalin (ENK), substance P (SP) and bombesin (BOM) were distributed. These immunoreactive fibers disappeared after the total denervation of the IMG. After the application of Fast Blue into the IMG or distal stumps of transected lumbar colonic and hypogastric nerves, retrogradely labeled neurons occurred in the myenteric plexus with increasing density along the distal colon and rectum, and were immunoreactive for VIP, DYN, CGRP, ENK, SP or BOM. Double immunostaining of nerve fibers in the distal stumps of the ligated colonic and hypogastric nerves revealed the presence of viscerofugal fibers containing VIP with DYN and/or CGRP and those containing ENK with SP and/or BOM. These results demonstrate for the first time that the efferent limb of the canine intestino-intestinal reflex arch via the IMG consists of Calb-immunoreactive ganglion neurons projecting to the longitudinal muscles in addition to the enteric plexus of the lower intestine and also of NPY-immunoreactive ganglion neurons projecting to the intestinal blood vessels, and that the afferent limb is composed of at least two discrete groups with different peptide contents, i.e., myenteric neurons containing VIP with DYN and/or CGRP and those containing ENK with SP and/or BOM. PMID- 9412743 TI - The cardiac internuncial cell in Carassius auratus longsdorffii and other 19 teleost species: a fine-structural study. AB - Thin-section studies of the sino-auricular border region in the hearts of 20 species of teleost fish revealed the cardiac internuncial cell (CIC), hitherto known only in Misgurnus, to occur also in 8 species (Acheilognathus lanceolatus, Carassius auratus longsdorffii, Cyprinus carpio, Girella punctata, Kareius bicoloratus, Rhodeus ocellatus ocellatus, Rhyncopelates oxyrhynchus, Tilapia nilotica), but to be absent in the other 12 species. There was an indication that the CIC was close in nature to myocardial cells because of the findings of Z band like structures, especially myosin-like thick filaments (ca. 15 nm thick) in the cytoplasm. However, the myosin-like thick filaments were not associated with the actin-like thin filaments (ca. 5 nm thick) or Z band-like structures. Interestingly, the thick filaments showed considerable variation in their occurrence among individuals of a species (Carassius auratus longsdorffii); they were scarce in all of the CICs (Type I) observed in 12 out of 15 individuals studied, and very numerous in all of the CICs (Type II) in the remaining 3 individuals. All of the CICs in Kareius bicoloratus, Girella punctata, Rhodeus ocellatus ocellatus and Rhyncopelates oxyrhynchus were likely to be Type I and all of the CICs in Acheilognathus lanceolatus, Cyprinus carpio and Tilapia nilotica, Type II. PMID- 9412744 TI - Occurrence of subtypes of gustatory cells in cat circumvallate taste buds. AB - Transmission-electron microscopy of cat taste buds confirmed that, as in other mammals, each taste bud comprised four distinct types of cells: Type I, Type II, Type III (gustatory), and Type IV (basal) cells. Gustatory cells made synaptic contacts with nerves to which synaptic vesicles were gathered. The following are the main findings on the cat gustatory cells: 1) The synaptic vesicles of gustatory cells were essentially all dense-cored in type; small clear vesicles, which usually are intermingled in other mammals, could not be found. 2) The vesicles were accumulated not only in the synaptic area but also in the basal cytoplasm. This implies endocrine and paracrine functions. 3) On the basis of the fine structure of the vesicles, two subtypes of gustatory cells were discriminated. A large part of the cells contained vesicles measuring about 180 nm in diameter, while a small part had smaller ones of about 100 nm in diameter. This is the first demonstration of a dual population of gustatory cells in mammals, suggesting different messenger substances utilized. PMID- 9412745 TI - Managing the excited skin syndrome: patch testing hyperirritable skin. AB - Inflammation-modulating phenomena (IMPs), humoral and cellular, fluctuate during the course of irritant and allergic contact dermatitis influencing irritability of the skin. The patch test procedure is a biological assay, a titration of responses of IMPs which can produce hyporeactivity or hyperirritability of the skin of patients who have dermatitis (PDs) and a single patch test is a 'snapshot' of the tempo of an evolving process. The excited skin syndrome (ESS) refers to hyperirritability from clinical and patch test dermatitis creating false-positive patch test reactions which are not reproducible when dermatitis and IMPs have subsided. During ESS, the threshold for irritancy decreases and irritant reactions increase. Patch test concentrations should be determined and ESS investigated in PDs having enhanced IMPs, not in 'normal' individuals, and if a patch test result is important to a patient the test should be performed more than once. Variable reproducibility is inherent in the patch test method, but ESS can be managed by appropriate testing and retesting, and search for relevance. PMID- 9412746 TI - National rates and regional differences in sensitization to allergens of the standard series. Population-adjusted frequencies of sensitization (PAFS) in 40,000 patients from a multicenter study (IVDK). AB - Sensitization rates to contact allergens vary between centers and are influenced by sex and age. Eliminating the latter 2 factors by standardization of data by age and sex, the present analysis addresses possible differences between centers remaining after elimination of these confounders, and analyzes other factors which might influence rates, e.g., the MOAHL index. Overall standardized rates were well within the range reported in previous studies and may be regarded as representing the rates of the "patch test population" in Central Europe (e.g., nickel sulfate 12.9%, fragrance mix 10.5%, balsam of Peru 7.3%, thimerosal 5.6%). For this analysis, data of those departments which contributed more than 2000 patients, or of those with extreme proportions concerning sex, age and occupational cases were selected. Patients from these 10 departments differed considerably with regard to the items of the MOAHL index and with regard to standardized rates. The items of the MOAHL index proved to be suitable for describing different patch test populations and for explaining some differences between centers. Only 'atopic dermatitis' seems to have little influence on (standardized) rates. Face dermatitis is not yet represented in the MOAHL index, but should be included, together with age > 40 years, in an extended index (acronym: MOAHLFA). Regional allergen exposure (with striking differences between East Germany, West Germany and, to a lesser extent, Austria) seems to have a great influence on the sensitization pattern observed in a department. In addition, sociological factors may influence sensitization rates, which is exemplified by high rates of nickel allergy in a socially defined subgroup. Future studies should focus on these factors, as well as on factors concerning patch test practices and quality control. PMID- 9412747 TI - Allergy to phosphorus sesquisulfide: 0.5% pet. is a more appropriate concentration for patch testing. AB - Patch testing was performed with phosphorus sesquisulfide P4S3 in 2 groups containing equal numbers of patients using different concentrations (0.5% P4S3 in pet. and 1% P4S3 in pet., the usual suggested test concentration as recommended by the International Contact Dermatitis Research Group). We found that there was a statistically significant increase in the number of clinically irrelevant irritant reactions in the group tested with the concentration (chi 2 = 16, p < 0.0004). We recommend that patch testing with phosphorous sesquisulfide should be at a reduced concentration of 0.5% pet. PMID- 9412748 TI - Irritancy of antiseptics tested by repeated open exposures on the human skin, evaluated by non-invasive methods. AB - The aim of the study was to test the irritancy of 6 antiseptics in an open exposure model. The following agents were tested in their normal use concentrations using open exposures, 2x daily for 4 days in 20 subjects: chlorhexidine 4% (CH), chlorhexidine 0.5% in ethanol 70% (CE), ethanol 70% (ET), iodine 1% in ethanol 70% (IE), povidone-iodine 10% (PI) and sodium hypochlorite 0.25% (SH). Responses were evaluated by visual scoring, subjective irritancy scoring, stratum corneum hydration (Corneometer), transepidermal water loss and laser Doppler flowmetry. Exposure to SH had to be discontinued after 4 applications because of severe subjective irritation. The same held true for IE (7 applications), whereas the other agents were exposed 8x. All evaluation methods showed SH to be significantly more irritating than IE, which was in turn more irritating than CH, CE, ET and PI. Thus, it can be concluded that CH, CE, ET and PI were non-irritating in this open exposure model. PMID- 9412749 TI - Visual assessment of human skin irritation: a sensitive and reproducible tool. AB - Human volunteer studies of skin irritation have been carried out for decades, both for research and for safety evaluation purposes. For the majority of this time, and consequently for the majority of the studies, assessment of the skin reactions has been made visually. Typical endpoints include erythema, oedema, dryness and scaling, some or all of which would be rated on a simple scale, eg 0, +/-, +, ++, + + +. Such approaches can be criticized as subjective, of poor reproducibility, lacking in sensitivity and highly variable between observers and/or institutions. In consequence, instrumental methods of assessment have been strongly promoted and do indeed offer several advantages, not least their objectivity. However, it is possible to use the human eye, which is a very sensitive tool, to make detailed, accurate and reproducible descriptions of skin irritation reactions. To achieve this, it is necessary to give prolonged and thorough training to each observer. In this paper, 3 examples of human volunteer studies, in which different pairs of trained observers independently carried out double blinded scoring of the irritation reactions, are reported. The grading patterns produced were almost identical; statistical analysis showed that properly trained observers are in fact able to reliably measure a grade of erythema to within +/- 1 on a 10 point scale; 97.6% of scores were within 2 grade points on this scale. These results provide evidence that visual scoring can be sensitive, reliable and reproducible within a testing institution. PMID- 9412750 TI - Contact and photocontact sensitivity to sunscreens. Review of a 15-year experience and of the literature. AB - This review summarizes published and unpublished data of our 15-year experience with sunscreen allergy and photoallergy. From 1981-1996, 402 patients with suspected clinical photosensitivity were patch and photopatch tested with the commercial sunscreens and facial cosmetics that they had used and with chemical UV absorbers, fragrance materials, preservatives, and emollients. 80 patients (20%) (28 men, 52 women) demonstrated allergic and/or photoallergic contact dermatitis to 1 or more UV absorber(s). In 47 patients with photodermatoses or photo-aggravated dermatoses and in 33 subjects with normal photosensitivity, 91 allergic and 84 photoallergic reactions to UV filters were observed. Over the years sunscreens were added to the test series, which since 1989 comprised the following 10 UV absorbers and which induced allergic (a) and photoallergic (pa) reactions (number, type of reaction): 4 UVA absorbers--isopropyldibenzoylmethane (30a/32pa); butyl methoxydibenzoyl-methane (15a/13pa); benzophenone-3 (3a/9pa); benzophenone-4 (0a/0pa); and 6 UVB absorbers--PABA (2a/2pa); octyl dimethyl PABA (1a/2pa); methylbenzylidene camphor (32a/5pa); octyl methoxycinnamate (3a/4pa); isoamyl p-methoxycinnamate (4a/10pa); and phenylbenzimidazole sulfonic acid (1a/7pa). The frequent (photo)sensitization to isopropyldibenzoylmethane was the reason that its production was discontinued in 1993. 47 patients reacted to fragrance materials, 11 to preservatives and 2 to lanolin alcohol. These constituents were contained in the commercial sunscreens and cosmetics that they had used. Continuous revision of the UV absorber photopatch test series was necessary to be closer to the real frequency of exposure and of reported (photo)allergy to newer sunscreens. Clinicians should consider contact and photocontact allergy, especially in patients with photodermatoses and photo aggravated dermatoses, and they should perform photopatch testing. Once the culprit has been identified, its INCI (International Nomenclature Cosmetic Ingredients) designation should be given to the patient, who must be warned to avoid products containing the (photo)allergen. PMID- 9412751 TI - Contact urticaria due to aescin. PMID- 9412752 TI - Delayed hypersensitivity to silicon causing gingival hyperplasia. PMID- 9412753 TI - Clinical atopy score and TEWL are not correlated in a cohort of metalworkers. PMID- 9412754 TI - Contact allergy to musk moskene in a perfumed moisturizing cream. PMID- 9412755 TI - An outbreak of contact dermatitis from Japanese lacquer tree. PMID- 9412756 TI - Active sensitization to p-aminodiphenylamine. PMID- 9412757 TI - Allergic contact dermatitis from a diisocyanate in wool processing. PMID- 9412758 TI - Contact dermatitis from moss in a forestry worker. PMID- 9412759 TI - Patch testing with low concentrations of budesonide detects contact allergy. PMID- 9412760 TI - Ketoprofen-induced connubial photodermatitis. PMID- 9412762 TI - Pigmented contact dermatitis of the lips from a lipstick. PMID- 9412761 TI - Colophony in topical traditional Chinese medicaments. PMID- 9412763 TI - Trends in the results of patch testing to standard allergens over the period 1984 1995. PMID- 9412764 TI - Protein contact dermatitis caused by Anisakis simplex. PMID- 9412765 TI - Contact dermatitis and stomatitis due to amine fluoride. PMID- 9412766 TI - Allergic contact dermatitis from rosemary (Rosmarinus officinalis L.). PMID- 9412767 TI - Maculopapular and urticarial eruption from cetirizine. PMID- 9412768 TI - Delayed-type reactivity to calcipotriol without cross-sensitization to tacalcitol. PMID- 9412770 TI - Allergic contact dermatitis from ophthalmics: update 1997. PMID- 9412771 TI - Therapy and clinical trials. PMID- 9412772 TI - Serum low-density lipoprotein cholesterol response to modification of saturated fat intake: recent insights. AB - Two lines of investigation have provided important insights into dietary therapy of hypercholesterolemia. Studies have confirmed the efficacy of the US National Cholesterol Education Program Step II diet in lowering serum LDL-cholesterol concentrations and demonstrate that wide variability exists in responsiveness to this diet. Other studies indicate that, contrary to expectations, cholesterol raising saturated fatty acids are not limited to long-chain saturates, but also include medium-chain and very long-chain saturated fatty acids. PMID- 9412773 TI - Therapy for lowering lipoprotein (a) levels. AB - Elevated levels of lipoprotein (a), a complex between apolipoprotein (a) and LDL, indicate an increased risk of atherosclerosis and cardiovascular disease. The important genetic regulation of lipoprotein (a) is now relatively well understood, but the progress in finding ways of influencing plasma lipoprotein (a) has been disappointingly slow. Nicotinic acid, pharmacological doses of sex hormones and anabolic steroids, as well as LDL apheresis may provide sufficient lowering of lipoprotein (a) for the evaluation of clinical response. Presently, the most important aspect is to take an evaluated lipoprotein (a) level into consideration when deciding to institute particularly aggressive lipid lowering treatment in a given individual. PMID- 9412774 TI - Treatment of lipid disorders in non-insulin-dependent diabetes mellitus. AB - The basis for treatment of lipid disorders in patients with non-insulin-dependent diabetes mellitus is weight reduction by diet and exercise, and additional control of glycaemic condition with oral antidiabetics, alone or in combination with insulin. Hypercholesterolaemic, mildly hypertriglyceridaemic non-insulin dependent diabetes mellitus patients respond to cholesterol malabsorption caused by dietary sitostanol ester margarine, while long-term statin treatment of respective coronary patients significantly lowers the recurrence of coronary events, in addition to improving the lipid disorder. However, no information is available concerning the preventive effect of long-term improvement of lipid disorders in non-insulin-dependent diabetes mellitus patients without coronary heart disease, or in patients with the 'classical' type of diabetic lipid disorder, that is, hypertriglyceridaemia with low HDL and normal-low LDL cholesterol levels. In this group of patients, beneficial lipid effects can be obtained (although perhaps not normalization) with fibrates alone or, especially, in combination with current statins. PMID- 9412775 TI - Stabilization of atherosclerotic plaque during lipid lowering. AB - Lipid lowering therapy leads to a great reduction of cardiovascular complications, but has almost no effect on the degree of stenosis of coronary arteries. These findings have lead to a new paradigm of coronary artery disease, i.e. clinical prognosis is not only determined by the extent of a single stenosis, but mainly by the number and structure of atherosclerotic plaque. Rupture of an instable or vulnerable plaque, characterized by a large lipid-rich central core, inflammatory cells, and a thin fibrous cap, causes sudden thrombus formation and thereby acute coronary syndromes. There is accumulating evidence that cholesterol lowering can result in plaque stabilization and improvement of endothelial dysfunction, reviewed recently in this journal. Accordingly, this review focuses on new molecular mechanisms which provide evidence that reduction of cellular cholesterol activates novel transcription factors, called sterol regulatory element binding proteins, which can regulate not only the LDL receptor, thereby reducing plasma cholesterol levels, but also a diverse number of other genes. These gene regulatory events might link cholesterol lowering not only to cellular cholesterol metabolism, but also to triglyceride metabolism, cell differentiation and other events potentially stabilizing vulnerable plaque. PMID- 9412776 TI - Effects of lipid lowering therapy on progression of coronary and carotid artery disease. AB - Recent data have extended the benefit of lipid lowering therapy to patients with only mildly to moderately elevated LDL-cholesterol, which is typical of patients with coronary artery disease. Meta-analysis of clinical trials of statin therapy with similar sample sizes indicated that the LDL-cholesterol level on treatment was as good a predictor of angiographic benefit as was the percentage reduction in LDL-cholesterol. We review evidence that management of triglyceride-rich lipoproteins, HDL, fibrinogen, lipoprotein particle size, LDL-oxidation, and lipoprotein (a) may also favorably influence atherosclerotic progression. Angiographic and arterial ultrasound trials of lipid lowering therapy have demonstrated benefits on disease progression that are consistent with benefits on myocardial infarction, stroke, and death reported in larger, lengthier trials. PMID- 9412777 TI - New lipid lowering drugs and new effects of old drugs. AB - Old lipid acting drugs (fibrates, resins and niacin) continue to demonstrate morbidity and mortality benefits with variable efficacy and safety. Controlled trials have provided efficacy and safety data that support the use of statins as the first choice in the treatment and prevention of atherosclerosis. Knowledge of old and new mechanisms of action, optimal doses, pharmacokinetic behavior and drug interactions improve the safety and effectiveness of these hypolipidemic agents. PMID- 9412778 TI - Improving health outcomes without increasing costs: maximizing the full potential of lipid reduction therapy in the primary and secondary prevention of coronary heart disease. AB - To more efficiently reduce the risk of coronary heart disease with lipid lowering therapy, cost effectiveness analysis offers an important tool to best determine how to allocate inherently limited resources to improve the health of both individuals and society. Formal economic analysis of the most recent clinical trials with the 3-hydroxymethylglutaryl coenzyme A reductase inhibitors (statins) suggest that secondary prevention of coronary heart disease is very cost effective compared to existing treatment and prevention strategies. Primary prevention is also cost effective but has a much wider range of cost effectiveness depending on an individual baseline risk. Cost effectiveness can be better maximized in primary prevention by treating patients at the highest absolute risk of coronary heart disease. The debate about lipid lowering therapy will now shift from that of efficacy and safety, to that of cost and cost effectiveness. Defining the optimal treatment thresholds for intervention in primary prevention will become a major focus of investigation. PMID- 9412779 TI - Therapy and clinical trials. PMID- 9412780 TI - Bimonthly update: lipidology. Nutrition. PMID- 9412781 TI - Genetic analysis by chromosome sorting and painting: phylogenetic and diagnostic applications. AB - Chromosome sorting from fluid suspensions of metaphase chromosomes using a fluorescence-activated cell sorter has been used for a number of years to produce chromosome-specific genomic libraries and other reagents for chromosome mapping. Improved techniques for fluorescence in situ hybridisation and the amplification and labelling of sorted chromosomes using degenerate oligonucleotide-primed PCR have led to the widespread use of chromosome painting both for the resolution of complex chromosome aberrations and for the study of karyotype evolution by cross species reciprocal chromosome painting. The chromosomes of a large number of different species have been sorted and used to make chromosome-specific paints and already new data challenging results of earlier phylogenetic studies have been obtained. Sorted chromosomes provide the resource for multicolour chromosome analysis of all chromosomes simultaneously. Such reagents are now available for all human and mouse chromosomes and are proving particularly useful in the analysis of cancer chromosomes. PMID- 9412783 TI - mtDNA haplotype analysis in Finnish families with leber hereditary optic neuroretinopathy. AB - The mitochondrial DNA (mtDNA) sequence variation of 24 Finnish Leber hereditary optic neuroretinopathy (LHON) probands was characterized by sequencing and restriction endonuclease analyses. All LHON-associated substitutions and Caucasoid haplogroup-specific mutations were screened in the families. Analysis of the mtDNAs revealed that the Finnish LHON families have two unique features: an absence of the ND6/14484 mutation and a high number of families (10/24) without the primary mutations ND1/3460 and ND4/11778. Furthermore, the LHON families showed considerable mtDNA heterogeneity: among 24 families 22 haplotypes were detected. Overall, the haplogrouping of LHON families was similar to other European populations. However, the frequency of ND4/11778-positive families in haplogroup J was high, which may indicate that background mutations in this haplogroup together with the ND4/11778 primary mutation promote the penetrance of LHON. PMID- 9412782 TI - A single-base deletion in the 3'-coding region of glycogen-debranching enzyme is prevalent in glycogen storage disease type IIIA in a population of North African Jewish patients. AB - Glycogen storage disease type III (GSD III) is an autosomal recessive disease caused by the deficiency of glycogen-debranching enzyme (AGL). The overall incidence of the disease is about 1:100,000 life births in the USA; however, it is unusually frequent among North African Jews in Israel (prevalence 1:5,400, carrier prevalence 1:35). All North African Jewish GSD III patients examined have both liver and muscle involvement. While all patients showed the characteristic features related to the liver enzyme deficiency, the peripheral muscular impairment varied from minimal to severe, with neuromuscular involvement. A single mutation in the AGL gene, the deletion of T at position 4,455 (4,455delT) in homozygous form, was found in this patient population. The mutation 4,455delT results in the change of 17 amino acids at the carboxy terminus of the AGL protein (1,486-1,502) and truncation of the last 30 amino acids of the normal AGL 1,532 amino acids. The mutation appears to be ethnic specific as it was not seen in 18 patients of different ethnic origins. This is the first report of a mutation in the AGL gene affecting a considerable number of GSD III patients in a defined population. PMID- 9412784 TI - Deletion of 11 amino acids in tuberin associated with severe tuberous sclerosis phenotypes: evidence for a new essential domain in the first third of the protein. AB - Tuberous sclerosis complex (TSC) is an autosomal dominant disorder displaying a large spectrum of symptoms. Linkage studies have shown two loci, TSC1 in 9q34 and TSC2 in 16p13.3, to be involved in the disease. The TSC2 gene, composed of 41 exons, has been isolated and is shown to encode a protein, tuberin, from a 5.5-kb transcript. Mutation screening for both clinical diagnosis and identification of functional domains within the tuberin is in progress. In this study we identify a 33-bp in-frame deletion (1462del33) in the mRNA which segregates in two unrelated French families with severe TSC phenotypes. The corresponding 11 amino acids deletion (aa 482-492) is shown to result from two different splice site mutations at exon 14 and, when compared with the position of two previously described missense mutations, indicates a novel functionally important region of the protein. PMID- 9412785 TI - Y-chromosome STR loci in Sardinia and continental Italy reveal islander-specific haplotypes. AB - Six Y-linked tetranucleotide microsatellites were typed in a sample of continental Italians and Sardinians. Significant differences in allele distributions were found between peninsular Italy and the island. Patterns of distinct allelic associations were evident in Sardinia and in the mainland. STR haplotypes in a subset of Sardinian chromosomes were monophyletically related and indicated that additions/deletions of a single tetranucleotide unit had to sequentially occur within a historical time-scale (about 9,000 years). Assumptions on both the time elapsed since the peopling of the island and the number of mutational events led us to estimate (by three different methods) a rate of 2.7-11 x 10(-4) mutations per generation per locus--at the upper end of the range of values reported in the literature. PMID- 9412786 TI - FMR1 premutation allele (CGG)81 is stable in mice. AB - Fragile X syndrome is caused by an expansion of the CGG repeat present in the 5' UTR of the FMR1 gene. A lot has been elucidated about the genetics of the disease, but not much is known about the mechanisms involved in repeat instability. Transgenic animals with a premutation allele [(CGG)11AGG(CGG)60CAG(CGG)8] in the human FMR1 promoter were generated to study the inheritance of this repeat in mice. Three independent lines, B6, B7 and B29, in total 263 transgenic animals, were tested for repeat instability. In all meiosis and mitosis tested, the repeat inherited stably. This suggests that other factors might be important in repeat (in)stability. PMID- 9412787 TI - Integrated radiation hybrid and yeast artificial chromosome map of chromosome 9p. AB - A panel of 93 radiation-reduced hybrids have been screened using PCR amplification and oligonucleotide primers for sequence-tagged sites (STSs) specific for 114 single-copy loci mapping to the short arm of chromosome 9. An x ray dose of 6,000 rads gave an average retention frequency of approximately 23%. We have constructed a framework map containing 31 markers ordered by analyzing coretention patterns, with support for the order greater than 1,000:1. In addition, we have placed the remaining markers which could not be mapped to a single interval with this support to a range of intervals on the framework map. The STS oligonucleotide primers used in the construction of the radiation hybrid (RH) map have been used to isolate and order yeast artificial chromosomes (YACs) assigned to 9p identified from the CEPH megaYAC library. Eighty-nine STS markers have screened positive with at least one YAC. A total of 88 individual YACs (with an average size of 0.9 MB) have been placed on the map in a series of contigs and in some cases mapped cytogenetically by fluorescence in situ hybridization. Additionally, the YAC information has been used in conjunction with the RH framework placements to generate an integrated map containing 65 loci including 51 uniquely positioned markers, with an average resolution of 0.79 Mb. PMID- 9412788 TI - Trisomy first, translocation second, uniparental disomy and partial trisomy third: a new mechanism for complex chromosomal aneuploidy. AB - A 2-year-old, short, microcephalic and developmentally retarded boy revealed a pattern of multiple minor anomalies, hypospadias and a dysplastic right kidney. Maternal age at delivery was 41 years. His karyotype showed two cell lines, one apparently normal, the other with a 1p+ chromosome. FISH examinations showed that the segment attached to 1p was from chromosome 16, and molecular investigations disclosed maternal heterodisomy 16, except for the segment (16)(pter-->p13.1) for which there was mosaicism between trisomy and uniparental disomy (UPD). Most likely, the zygote was trisomic for chromosome 16 due to a maternal meiosis I nondisjunction; a somatic rearrangement would have then occurred at an early postzygotic stage whereby a segment of the paternal chromosome 16 was translocated onto 1p. Subsequently, the paternal chromosomes 16 and 16p- had been lost in the normal and the translocation cell line, respectively. The chromosome aberration was detected secondary to the disclosure of maternal UPD 16 because of the demonstration of a paternal band at several loci on distal 16p. This case shows that chromosome aberrations may be formed in a more complicated manner than primarily assumed. Hence, the phenotype might also be due to underlying factors such as UPD or undetected mosaicism in addition to the more obvious implications of the chromosome rearrangement itself (e.g. partial trisomy). PMID- 9412789 TI - A xanthomatosis-susceptibility gene may exist in a Syrian family with familial hypercholesterolemia. AB - Familial hypercholesterolemia (FH) is an autosomal-dominant inherited disorder characterized by high serum low-density lipoprotein (LDL)-cholesterol concentrations, xanthoma formation, and premature atherosclerosis. Homozygous individuals die of vascular disease as children or young adults; heterozygous persons are at high risk for premature cardiovascular death. Mutations in the LDL receptor gene are responsible for FH. We studied 49 members of a consanguineous Syrian kindred containing 6 homozygous individuals from the same pedigree. Half of the homozygotes had giant xanthomas, while half did not, even though their LDL cholesterol concentrations were elevated to similar degrees (> 14 mmol/l). Heterozygous FH individuals from this family were also clearly distinguishable with respect to xanthoma size. We performed DNA analysis and were successful in identifying a hitherto not described mutation in this family's LDL receptor. DNA sequence analysis of the LDL-receptor gene revealed a T to C substitution at nucleotide 1,999 in codon 646 of exon 14. We next conducted a segregation analysis, which suggests that a susceptibility gene may explain the formation of giant xanthomas in this family. We raise the hypothesis that the appearance of giant xanthomas in this FH family is controlled by a second gene acting in an autosomal-dominant or recessive fashion. Elucidation of this 'xanthoma' gene may shed additional light on LDL-cholesterol deposition. PMID- 9412790 TI - The mouse Necdin gene is expressed from the paternal allele only and lies in the 7C region of the mouse chromosome 7, a region of conserved synteny to the human Prader-Willi syndrome region. AB - Prader-Willi syndrome (PWS) is a neurogenetic disorder resulting from the loss of paternal expression of gene(s) localized in the 15q11-q12 region. A new human gene encoding a putative protein with high homology to the mouse NECDIN protein has recently been characterized and mapped to chromosome 15q11-q12. It is expressed from the paternal allele only, suggesting its potential involvement in PWS. We now report the localization of the mouse Necdin gene in a region of conserved synteny to the human PWS region. We demonstrate the paternal specific expression of Necdin in the mouse central nervous system, and show that parental alleles display a differential methylation profile in the coding region. Finally, fluorescence in situ hybridization analysis reveals an asynchronous pattern of replication at the Necdin locus. These results clearly demonstrate imprinting of the mouse Necdin gene. Mouse models will be powerful tools in the study of human PWS phenotype and imprinting mechanisms. PMID- 9412791 TI - Genetic variation at the apoB 3'hypervariable region in a Serbian population. AB - We investigated common length polymorphism caused by a variable number of tandem repeats in the hypervariable region located at the 3' end of the human apolipoprotein B gene in 696 Serbian (Belgrade area) unrelated individuals of both genders. After using the polymerase chain reaction to amplify this polymorphic region, 17 different alleles, containing 22-54 repeats, were distinguished. The bimodal distribution and the heterozygosity index (average 0.71) obtained in both genders are similar to those reported for other Caucasian populations. However, the HVE34 allele was found to be the commonest in both female and male samples. There was also a lower frequency of the HVE > 36 alleles than in other Caucasian populations studied. PMID- 9412792 TI - Further delineation of the critical region for noonan syndrome on the long arm of chromosome 12. PMID- 9412793 TI - Results of radical prostatectomy in men with locally advanced prostate cancer: multi-institutional pooled analysis. AB - OBJECTIVE: We investigated the disease-specific and metastasis-free survival rates in men with locally advanced (clinical stage T3) prostate cancer who were treated surgically. METHODS: A retrospective, multi-institutional pooled analysis of the results of surgical treatment in 345 men with clinical stage T3 disease was performed. Survival curves were generated using the Kaplan-Meier method. RESULTS: Among 298 evaluable patients, pelvic lymphadenectomy alone was performed in 56 men (19%), while 242 men (81%) underwent node dissection and radical prostatectomy. In total, 122 of 298 patients (41%) had nodal metastases and/or seminal vesicle tumor spread. Pathologically organ-confined disease was noted in 27 men (9%). The actuarial 10-year disease-specific and metastasis-free survival rates for all patients managed surgically were 57 and 32%, respectively. For patients with well, moderately and poorly differentiated tumors, cancer-specific survival rates at 10 years were 73, 67 and 29%, respectively. CONCLUSIONS: A large number of men with clinical stage T3 prostate cancer have advanced disease and are unlikely to achieve improved long-term survival with surgery alone. Although there may be a role for radical prostatectomy in selected patients with low to intermediate grade tumors, such treatment appears unlikely to result in long-term survival in men with high grade disease. A prospective study is necessary to determine the optimal treatment approach in men with locally advanced prostate cancer. PMID- 9412795 TI - A randomized comparison of the clinical and hormonal effects of two GnRH agonists in patients with prostate cancer. AB - OBJECTIVE: The aims of the study were (i) to compared the efficacy of the two long-acting GnRH agonists (GnRHa) triptorelin (Trp) and leuprolide (Leu) in men with prostate cancer and (ii) to assess the pattern of plasma testosterone levels following each injection of GnRHa. PATIENTS AND METHODS: 67 patients referred for prostate cancer not suitable for surgery were randomly allocated to two treatment regimens: 33 patients received 3.75 mg Trp i.m. at 4-week intervals for 3 months and 34 patients were treated with 3.75 mg Leu s.c. at the same rhythm of administration for 3 months. RESULTS: Clinical data at entry and assessed monthly during follow-up did not differ between the two groups. Plasma prostate-specific antigen (PSA) and testosterone were measured before, 24 and 72 h after each injection of GnRHa. During treatment, PSA dropped similarly in both groups. By month 2, testosterone was < 1.0 nmol/l in 77 and 48% of patients treated with Trp and Leu, respectively (p = 0.02). 24 and 72 h after GnRHa injection, 77 (Trp) and 56% (Leu) of patients had testosterone < 1.0 nmol/l (p < 0.05). CONCLUSIONS: The second and third injections of GnRHa were not followed by a significant increase in testosterone. Trp induced a higher decrease in testosterone than did Leu. The implications in terms of survival should, however, be studied in a larger and longer study. PMID- 9412794 TI - Flutamide versus orchidectomy in the treatment of metastatic prostate carcinoma. AB - PURPOSE: To compare in a randomized clinical trial the therapeutic efficacy of the nonsteroidal antiandrogen flutamide 250 mg tid to testicular androgen suppression by orchidectomy in patients with metastatic prostate cancer. PATIENTS AND METHODS: Between 1989 and 1991, 104 patients aged 74 +/- 8 years with newly diagnosed metastatic prostate cancer, an ECOG performance status 0-2 and no prior hormone manipulation or chemotherapy, were randomized to receive flutamide 250 mg tid (54 patients) or orchidectomy (50 patients). Patients were evaluated at entry and at months 3, 6, 12, 18 and 24. The primary endpoint was duration of progression-free survival, progression being defined as an increase in PSA> 50% over the nadir value at 2 consecutive months or a single PSA rise > 50% over the nadir value with another objective parameter. At progression, the treatment was left to the discretion of the attending urologist. RESULTS: 16 patients (10 flutamide, 6 orchidectomy) are not evaluable. 86 had a minimum follow-up of 36 months, 36/42 and 41/44 have progressed in the orchidectomy and flutamide group with a time of failure of 419 and 496 days (p = 0.32); median time to progression was almost identical in both groups (370 vs. 396 days p = 0.9); overall survival at 69 months irrespective of treatment at relapse was identical in both groups. Side effects were dominated by gynecomastia, hot flushes in both groups, breast tenderness and diarrhea in the flutamide group. Overall, 4 (10%) of the patients in the flutamide group withdrew from therapy because of side effects. The impact of flutamide on sexual potency was not assessed because of the advanced age of the patients. Serum testosterone rose by 50% over baseline level at month 3 to plateau at 25% over baseline level at month 12. CONCLUSION: Although affected by the lack of a clear statistical power due to the small number of patients in each arm, this study shows that in spite of a constant elevation of serum testosterone (25% over baseline) flutamide 250 mg tid may be a reasonable alternative to castration in highly selected patients with well to moderately differentiated low volume metastatic prostate cancer and wishing to avoid the side effects of androgen deprivation, provided they are closely monitored and ready to switch to standard androgen deprivation in the presence of untolerable side effects or suboptimal treatment efficacy as assessed by the inability to achieve a low PSA nadir. PMID- 9412796 TI - Long-term outcome of conservative therapy in men presenting with voiding symptoms and prostate cancer. AB - OBJECTIVE: To investigate the outcome of conservative therapy in men presenting with voiding symptoms and prostate cancer. METHODS: A consecutive series of 186 men presenting with voiding symptoms and prostate cancer were treated with transurethral resection (TUR). Examination of the resected tissue revealed 70 nonpalpable prostate cancers and confirmed the clinical suspicion of prostate cancer in 116 palpable tumors; 47 tumors were well differentiated, 87 intermediate and 52 poorly differentiated. Bone scan indicated metastasis in 24 men, all asymptomatic. The men were followed and underwent orchidectomy if symptoms of generalized disease appeared. RESULTS: After a follow-up of 13-21 years, 172/186 (92%) of the men had died, with 80/186 (43%) of the men dying of prostate cancer. The mean life expectancy was 6.3 years (confidence interval 5.4 7.1) compared with 10.2 years of an age-matched control group. In a subgroup of men with clinically localized disease, 26/97 (27%) died of prostate cancer. These men had a mean life expectancy of 7.1 years (confidence interval 6.0-8.3). Tumor stage and grade were highly significant predictors for cause-specific survival in uni- and multivariate analysis. Death from prostate cancer continued to occur beyond 10 years after diagnosis at a decreasing rate. CONCLUSIONS: Patients with prostate cancer causing voiding symptoms at presentation severe enough to necessitate TUR had a less favorable outcome than asymptomatic patients with prostate cancer in previously reported series, even when stratified for stage and grade. It is suggested that voiding symptoms at diagnosis are a putative prognostic factor in prostate cancer. PMID- 9412797 TI - Value of Ki-67 immunolabelling as a prognostic factor in prostate cancer. AB - OBJECTIVE(S): The aim of the study was to analyze the results of Ki-67 immunostaining in prostatic adenocarcinoma and to assess its prognostic value. METHODS: Clinical follow-up data was reviewed in 190 prostatic adenocarcinomas and the results of Ki-67 immunolabelling were correlated to standard prognostic factors and survival data of the patients. All stage and grade categories were included. RESULTS: Ki-67 expression correlated significantly with histological differentiation of tumors, indicators of cell proliferation, perineural growth, density of tumor-infiltrating lymphocytes and TM classification. In survival analysis, Ki-67 expression was able to discriminate patients into different prognostic groups (p = 0.0035). In a separate analysis including T1-2M0 tumors, Ki-67 immunolabelling was a significant predictor of survival (p = 0.0258). In Cox's multivariate analysis Ki-67 was an independent prognostic factor in the entire cohort. CONCLUSIONS: The results show that Ki-67 expression is a potentially useful prognostic factor in prostatic adenocarcinoma and it could be used as an additional criterion in defining a correct prognostic category in this malignancy. PMID- 9412798 TI - The correlation between prostate-specific antigen and age. Analysis of prostate specific antigen values from 4,846 Turkish men with symptomatic benign prostatic hyperplasia. AB - OBJECTIVE: To analyze serum prostate-specific antigen (PSA) levels of men proven to have benign prostatic hyperplasia (BPH) and to document any correlation between PSA, age and resected or enucleated prostate tissue. PATIENTS AND METHODS: Serum PSA values, age and weight of specimens of 4,846 men who underwent prostatectomy in the Ankara region between January 1, 1991, and December 24, 1995, were reviewed retrospectively. RESULTS: Serum PSA values correlated directly with age (Pearson's r = 0.45, p < 0.00001). The mean PSA values of men in each decade of age differed significantly (p < 0.0001) from all other decades. However, the correlation between the weight of the prostatectomy specimen and age or PSA did not reach statistical significance. CONCLUSION: The effect of age on PSA is evident in men with symptoms of BPH. The existence of symptomatic BPH should also be considered together with age when interpreting PSA levels. PMID- 9412799 TI - Evaluation of transrectal voiding ultrasonography in men with micturition difficulties without apparent organic obstruction of the lower urinary tract. AB - OBJECTIVE: To examine whether transrectal voiding ultrasonography (TRVUS) can evaluate voiding movement in men with dysfunctional voiding. METHODS: Ninety-nine consecutive men complaining of voiding difficulties without benign prostatic hyperplasia, prostatic cancer, severe bladder neck contracture and urethral stricture received uroflowmetry and TRVUS. Those who had abnormal findings on both uroflowmetry and TRVUS underwent subsequent cystometry combined with electromyography (EMG) to confirm the presence of dysfunctional voiding. RESULTS: Uroflowmetry indicated abnormal findings in 31 of the 99 patients, and TRVUS demonstrated abnormal movements of the posterior urethra during voiding in all of these 31 patients and 11 of the other 68 patients whose uroflowmetry did not indicate abnormality. TRVUS findings of the former 31 were divided into type E (the external urethral sphincter closed or intermittently opened while the bladder neck manifested an opening movement of > 7 mm during voiding in 20) and type I (both the bladder neck and external urethral sphincter manifested an intermittent movement of < 7 mm in 11). Subsequent cystometry combined with EMG in the 31 patients who had abnormal findings on both uroflowmetry and TRVUS revealed overactivity of the external urethral sphincter (OS) and underactivity of the detrusor (UD) in 85 and 35% of type-E group and 55 and 73% of type-I group, respectively. Type E included significantly more OS without UD than type I (65 vs. 18%; p = 0.0233). All of type-E (20/20) and 91% of type-I (10/11) patients had voiding difficulty which resulted from either OS or UD, while a very limited number of patients (4/31) manifested neurological symptoms such as paraplegia except for voiding difficulties. CONCLUSIONS: Both uroflowmetry and TRVUS are easy and useful methods to evaluate dysfunctional voiding in men, especially when neural disorders or organic obstruction of the lower urinary tract are not apparent. PMID- 9412800 TI - Bard BTA test compared with voided urine cytology in the diagnosis of recurrent bladder cancer. AB - OBJECTIVE: To compare the Bard BTA (bladder tumour antigen) test with voided urine cytology (VUC) in the diagnosis of recurrent bladder cancer (BC). METHODS: Urine specimens for the BTA test and VUC were collected on the same day as before cystoscopy from patients undergoing routine surveillance cystoscopy for recurrent BC. The pathologists performing VUC were blinded to the results of the BTA test. RESULTS: BC was identified by cystoscopy and biopsy in 39 of 164 study participants. The overall sensitivities of the BTA test and VUC were 54 and 28%, respectively (p < 0.05). The BTA test was more sensitive than VUC for all tumour stages and grades. For > or = T2 tumours and grade 3 tumours, respectively, the difference was statistically significant (p < 0.05). The specificities of the BTA test and VUC were 92 and 97%, respectively. Both a false-positive BTA test and VUC were found to predict recurrence. CONCLUSION: The BTA test is equal or superior to VUC in the detection of BC in patients undergoing routine surveillance for recurrent BC. PMID- 9412801 TI - Bladder biofeedback for the treatment of refractory sensory urgency in adults. AB - OBJECTIVE: Evaluation of bladder biofeedback in patients with sensory urgency refractory to medication and classical bladder training. METHODS: In 12 such patients a non-electronic technique of bladder biofeedback was used. Ambulatory treatment sessions were done once a week for 4 weeks, as opposed to every 14 days, with a mean total treatment period of 8 weeks. Micturition charts were completed before, during and 14 days after treatment by the patient. Before starting, patients had a mean of 15.8 micturitions/day, 2.3 micturitions/night, and a mean functional bladder capacity of 96 ml (26-172 ml). Urodynamic investigation showed a low-capacity bladder, hypersensitivity in some patients, and normal urodynamic parameters in others. RESULTS: At the end of biofeedback, patients had a mean of 5.7 micturitions/day, 0.3 micturitions/night, and a mean functional bladder capacity of 296 ml (163-470 ml). The results 9 months after completing the treatment were unchanged. Quality of life improved substantially in all. CONCLUSIONS: Bladder biofeedback is a valuable treatment for sensory urgency refractory to classical treatment. PMID- 9412803 TI - Grade, local stage and growth pattern as prognostic factors in carcinoma of the penis. AB - OBJECTIVES: To assess the management of adenopathies and the determination of prognostic factors in the treatment of squamous carcinoma of the penis. MATERIALS AND METHODS: We reviewed 81 cases treated at our Institution between 1980 and 1994. In 62 cases, we review the pathologic samples according to the growth patterns proposed by Cubilla et al. RESULTS: We find that both the grade of cellular differentiation and the local stage of the tumor are statistically significant with respect to the presence of positive adenopathies. We obtain results similar to Cubilla's work and different survival rates for the patterns of tumor growth. CONCLUSIONS: On the basis of our results, we propose 3 differentiated follow-up and treatment groups. We suggest that the new growth pattern classification is useful as prognostic factor in carcinoma of the penis. We corroborate that prophylactic lymphadenectomy is indicated in tumors of vertical growth, while a conservative management should be considered in tumors of verrucous growth. PMID- 9412802 TI - Excision and anastomotic repair for urethral stricture disease: experience with 150 cases. AB - PURPOSE: To analyze the results of a series of end-to-end urethroplasties performed in our service from 1968 to 1995 and of the factors contributing to failure. MATERIAL AND METHODS: 150 men (mean age 35.9 years) with urethral stricture disease underwent excision of the stricture and end-to-end anastomosis; in 95 it was the first attempt at repair while in 55 it was a secondary attempt. Eighty-two patients (54.6%) had a trauma-related stricture; of them, 56 followed a pelvic ring fracture with posterior urethra distraction defect, 24 (16%) had inflammatory strictures, 26 (17.3%) iatrogenic, 9 (6%) congenital, and 9 (6%) of unknown etiology; 81 (54%) were located in the bulbous urethra, 9 (6%) in the penoscrotal junction and 2 (1.3%) in the penile urethra. Ninety-one (60.6%) of the strictures or obliterative defects measured between 1 and 3 cm, 42 (28%) less than 1 cm and only 16 (10.6%) more than 3 cm. A perineal approach was used in 138 of the cases, while combined abdominoperineal route was necessary in 12; of these, 5 were children. The follow-up has ranged from 6 to 168 months (mean 44.4). The results were classified as good, fair (some re-stricturing, not needing treatment) and poor (recurrence). RESULTS: One hundred and twenty-six (84%) good outcomes, 10 (6.6%) fair, 14 (9.3%) poor. The factors influencing success or failure were: (1) primary or secondary character of the operation; (2) etiology; (3) length, and (4) location. Postoperative early complications consisted of 2 wound infections and 2 hematomas; as late complications, 1 chordee, 2 incontinence, 7 erectile dysfunction (in previously potent patients). The 14 patients considered as failures were operated again, all successfully; in 4 of them, a repeat excision and end-to-end anastomosis was performed, elevating the final success rate of the series to 93.3%. CONCLUSION: Excision and anastomotic repair represent the optimal mode of stricture repair for single lesions located from the penoscrotal junction to the membranous part of the urethra. PMID- 9412804 TI - Preliminary results of muscle cuff cervicoplasty in the ewe for the treatment of urinary incontinence. AB - OBJECTIVE: Muscle cuff cervicoplasty consists of dissecting a strip of the superior slip of the levator ani muscle, by sectioning it flush with the symphysis pubis, and applying it like a cuff around the urethra below the bladder neck. METHODS: We conducted a preliminary study in 12 ewes, including preoperative urodynamic assessment, intravenous urography with retrograde cystography on day 15, and another urodynamic assessment 1 and 3 months after the operation. Two animals died, but the other 10 animals were able to be evaluated. An increase in the urethral functional length with an infracervical plateau was revealed on all postoperative urethral profiles. Only transrectal electrostimulation, performed on the last 5 ewes, demonstrated an elevation of the closure pressure by 5-10 cm H2O. At sacrifice of the animals at 4 months, in situ stimulation of the muscle strip induced muscle contraction in 7 of 10 cases. Histological examination demonstrated persistence of muscle fibers in the cuff, despite the presence of marked fibrosis. RESULTS: The preliminary results are encouraging: urinary continence was improved with a minimal risk of retention, and cervicoplasty appears to retain its contractile activity, suggesting the possibility of a long-term sphincteric and proprioceptive action. CONCLUSION: This technique could be proposed in women suffering from urinary stress incontinence with sphincter hypoactivity. PMID- 9412805 TI - Latex thimble for needle suspension procedures. AB - OBJECTIVE: To present a latex finger tip protector designed to reduce the risk of finger tip injury during needle suspensions for correction of stress urinary incontinence. MATERIAL AND METHODS: The latex finger tip protector is placed over the gloved tip of the index finger. The suture carrier is kept in contact with the protected finger tip during passages through the abdominal fascia and retropubic space. RESULTS: We have been using the finger tip protector for 2 years and in more than 50 modified Peyrera bladder neck suspensions. We did not experience any finger injury and the procedure was found to be more comfortable for the surgeon. CONCLUSION: The latex finger tip protector is useful whenever the finger is used to control the passage of the ligature carrier through the retropubic space. PMID- 9412806 TI - Color Doppler imaging in the diagnosis of the acute scrotum. AB - OBJECTIVES: To determine the value of color Doppler imaging (CDI) in the diagnosis of acute scrotum. MATERIALS AND METHODS: We evaluated 102 consecutive patients referred to our institution with scrotal pain. All patients were evaluated by physical examination, ultrasound with stethoscopic Doppler followed by CDI. Radionuclide scanning was performed only in 14 patients. Patients with a diagnosis of testicular torsion by CDI underwent surgical exploration. Patients with CDI diagnosis of epididymorchitis were treated with intravenous antibodies and underwent repeat CDI evaluation after treatment. RESULTS: Of the 102 patients evaluated ages 11-44, CDI diagnosed 18 patients with testicular torsion and 78 patients with epididymorchitis. Cases of testicular torsion were found to have absent flow on CDI. CDI findings of normal or increased flow were present in all patients with epididymorchitis. All cases of torsion were confirmed on surgical exploration, 11 patients underwent orchidopexy and 7 patients underwent orchiectomy. No cases of testicular atrophy were encountered on long-term follow up in patients with epididymorchitis. Diagnoses in the remaining patients were testicular tumor 1, testicular abscess 2, scrotal hematoma 2 and incarcerated hernia 1. CDI was found to be 100% sensitive and 100% specific in the diagnosis of the acute scrotum. CONCLUSION: We found CDI to be more accurate and reliable than physical examination in conjunction with gray-scale ultrasound and Doppler stethoscopic examination in the differential diagnosis of the acute scrotum. CDI is practical and can be performed in a rapid manner. CDI should become an integral part of the urologist's armamentarium in the differential diagnosis of the acute scrotum. PMID- 9412807 TI - Tamsulosin 0.4 mg once daily: tolerability in older and younger patients with lower urinary tract symptoms suggestive of benign prostatic obstruction (symptomatic BPH). The European Tamsulosin Study Group. AB - OBJECTIVES: To compare the safety and tolerability of tamsulosin 0.4 mg once daily in younger (< 65 years) and older (> or = 65 years) patients with lower urinary tract symptoms (LUTS) suggestive of benign prostatic obstruction (BPO). METHODS: In a retrospective analysis of two European double-blind, randomized, placebo-controlled trials, safety was assessed in 574 younger or older patients treated with tamsulosin or placebo for 12 weeks. RESULTS: The incidence of adverse events, drug-related adverse events, serious adverse events and discontinuations due to adverse events was similar in older and younger tamsulosin-treated patients and was not significantly different from placebo. Although abnormal ejaculation was slightly more common in younger than older men receiving tamsulosin, the difference was not statistically significant from the placebo groups in both age groups. The incidence of adverse events possibly associated with vasodilation in tamsulosin-treated younger and older patients was 8.4 and 4.2%, respectively; these were comparable with the values for placebo treated patients: 7.5 and 6%, respectively. Baseline systolic blood pressure was higher in older than younger patients, but there were minimal changes in blood pressure or pulse rate in tamsulosin- or placebo-treated patients in either age group. CONCLUSIONS: Tamsulosin is well tolerated and suitable for use in older and younger patients with LUTS suggestive of BPO (symptomatic BPH). PMID- 9412808 TI - Retrograde endoureteropyelotomy with the holmium:YAG laser. Initial experience. AB - OBJECTIVE: To evaluate the safety and efficacy of the holmium:YAG laser in the management of pelviureteric junction (PUJ) obstruction. PATIENTS AND METHODS: Between November 1994 and February 1996, 8 patients with 5 primary and 3 secondary PUJ obstructions underwent retrograde ureteroscopic holmium:YAG laser endopyelotomy. A 320- to 365-micron (Slimline, Coherent) fibre was used to deliver the laser energy. A posterolateral endopyelotomy was performed under visual monitoring. All patients had a ureteral stent left in place postoperatively for 6 weeks and a renogram was performed after 3 months. The mean follow-up was 12.4 months (3-24 months). RESULTS: Almost bloodless, accurate and layered division of the PUJ was easily performed. The average procedure time was 37.5 min. Our overall success rate was 87.5%. CONCLUSION: The holmium:YAG laser is a safe, reliable laser with adequate coagulative properties and precise cutting abilities. This allows a 'bloodless' operative field for controlled, accurate and safe endoscopic division of the PUJ. PMID- 9412809 TI - Short- and mid-term outcome of different types of one-stage hypospadias corrections. AB - OBJECTIVE: This study is a retrospective analysis of the outcome of our one-stage hypospadias corrections. PATIENTS AND METHODS: In the period from August 1979 to January 1991, 316 boys less than 14 years of age underwent a one-stage hypospadias correction in our hospital. For 283 patients, data on the familial incidence, associated urological pathology, age at the time of operation, operation technique and follow-up were analyzed. RESULTS: The familial incidence in our study group was 9.4%, associated cryptorchidism was seen in 8.6%, kidney abnormalities in 1.8% and urethral valves in 0.4%. The majority (65.3%) of the children were operated on before the age of 3 years. The operation was performed according to Magpi, Mathieu, Devine-Horton, an onlay, and tube formation according to Duckett and the double-faced island flap procedure. Including all postoperative disturbances, urethral complications were seen in 18.1% and skin related complications in 4.6% of the patients. CONCLUSION: Even in experienced hands the overall complications rate is relatively high, and additional problems can be expected in the future. It is important to provide adequate information on the procedures and possible complication to the parents. The final long-term outcome after puberty has to be awaited. PMID- 9412810 TI - Membrano-bulbo-urethral junction stenosis. Posterior urethra obstruction due to extreme caliber disproportion in the male urethra. AB - OBJECTIVE: Based on 4 cases of infravesical obstruction due to extreme caliber disproportion between the posterior urethra and the penile urethra, a pathophysiological mechanism for this dynamic obstruction is given and endoscopic treatment is described. SUBJECTS: Four cases of membrano-bulbo-urethral junction (MBUJ) stenosis, seen between September 1995 and April 1996, are described. Two boys had previous successful valve resection but still showed extreme ballooning of the posterior urethra. The other 2 boys showed bladder instability on urodynamics and the male variant of the spinning top urethra on voiding cystourethrography (VCUG). RESULTS: All cases were successfully treated by endoscopic incision at the 12 o'clock position of the kink between the posterior and the penile urethra which is seen when the full bladder is expressed. Disproportion in the posterior urethra, seen on VCUG, together with bad urinary flow measured on uroflowmetry raise the suspicion of MBUJ stenosis. CONCLUSION: Although rarely seen, extreme caliber disproportion in the male urethra can cause obstruction. Ballooning of the posterior urethra, caused by urethral valves, bladder instability resisted by voluntary sphincter contraction or congenital posterior urethral dilatation, creates an obstructive kink in the urethra comparable to some obstructions in ureteropelvic junction stenosis. If suspicion of such a form of obstruction arises, cystoscopy during pressure on the full bladder is mandatory in order to see the obstruction, descending as a membrane from the vault of the urethra. PMID- 9412811 TI - Use of new nephrostomy catheter for treatment of renal neonatal candidiasis. AB - Systemic candidiasis with renal involvement is a rare but well-recognized complication during neonatal intensive care treatment. In addition to intravenous administration of amphotericin B, decompression of the renal pelvis and irrigation of the involved kidney with the same drug through a nephrostomy tube will provide a high concentration of antifungal agent with a flushing effect. This procedure is not always possible due to the small size of the neonatal kidneys. We have conceived a new percutaneous trocar nephrostomy which allows its application directly in an incubator without using X-rays during a single procedure. In 3 cases a bilateral percutaneous nephrostomy was performed directly in the incubator using a one-step ultrasonically guided maneuver under local anesthesia. The funguria was successfully eradicated in all cases. The availability of a nephrostomy trocar of small dimensions leads us to an improved renal approach in newborns. PMID- 9412812 TI - p53 and MDM2 in the development and progression of bladder cancer. AB - OBJECTIVE: Earlier investigations have demonstrated that inactivation of the p53 tumor suppressor gene might play a role in the development and progression of bladder cancer. Complex formation with the MDM2 oncogene product is one mechanism inactivating the p53 protein. Therefore, the MDM2 and the p53 protein were investigated to study potential interactions in bladder cancer. METHOD: 200 archival bladder tissue specimens from 92 patients were studied by immunohistochemistry using monoclonal antibodies DO-1 against p53 and IF2 against MDM2. RESULTS: No staining was observed for p53 or MDM2 in normal urothelium. Alterations of both genes were rare in dysplasia. p53 accumulation was observed in 27-44% of the tumor stages examined. MDM2 overexpression increased from 18% in carcinoma in situ to 49% in T1 tumors, but was present in only 22% of the advanced tumors. Alterations of both genes were more frequent in high-grade lesions. To investigate the prognostic impact of these alterations 61 patients with superficial bladder tumors were followed for at least 2 years (mean 51 months). Multivariate analysis demonstrated that multifocal disease and p53 accumulation were significantly correlated with tumor progression (p = 0.0099 and 0.0135). MDM2 overexpression alone had no prognostic significance. Patients with alterations of both genes had a very high risk of tumor progression (p = 0.0064). CONCLUSION: These results demonstrate a positive correlation between p53 accumulation and MDM2 overexpression in the progression of bladder cancer which may have prognostic value. PMID- 9412813 TI - Detection of telomerase activity in prostate cancer by needle biopsy. AB - OBJECTIVES: Re-expression of telomerase is believed to play an important role in immortalization and carcinogenesis. Thus, telomerase activity is considered to be a potentially useful diagnostic marker. We evaluated the potential diagnostic use of assaying telomerase activity in needle biopsy samples of prostate. METHODS: 114 were obtained from 38 patients with suspected cancer who underwent transrectal ultrasound-guided systematic prostate needle biopsy. Two samples were obtained per site for histological examination and telomerase assay. The activity was assayed using the telomeric repeat amplification protocol and correlated with histological findings. RESULTS: Of the total of 114 samples, 57 were obtained from 22 prostate cancer patients. Telomerase activity was detected in 66% (25/38) of the samples which were histologically confirmed to contain cancerous cells and in 11% (2/19) of the samples from adjacent noncancerous tissues. Of 22 patients with prostate cancer, 82% were positive for telomerase activity in one or more samples by needle biopsy, whereas 1 of 16 patients without histologically cancerous tissues was positive for the telomerase. CONCLUSIONS: The results indicate that telomerase activity in needle biopsy samples is a useful diagnostic marker of prostate cancer. PMID- 9412814 TI - Orthotopic reconstruction in a woman following cystectomy and cutaneous urinary diversion. The 1st reported case. AB - Orthotopic lower urinary tract reconstruction has become the procedure of choice in selected male and female patients at our institution following cystectomy with excellent functional results. A natural extension of the orthotopic neobladder is undiversion to the intact native urethra in patients who had previously undergone cystectomy and cutaneous urinary diversion. Undiversion has been successfully performed in selected male patients; however, to our knowledge, undiversion has not been reported in women. Herein, we present the 1st case of undiversion in a female patient who had undergone prior cystectomy and cutaneous urinary diversion. PMID- 9412815 TI - Skin changes in "screen dermatitis" versus classical UV- and ionizing irradiation related damage--similarities and differences. AB - An increasing number of persons say that they get cutaneous problems as well as symptoms from certain internal organs, such as the central nervous system (CNS) and the heart, when being close to electric equipment. A major group of these patients are the users of video display terminals (VDTs), who claim to have subjective and objective skin- and mucosa-related symptoms, such as pain, itch, heat sensation, erythema, papules, and pustules. The CNS symptoms are, e.g. dizziness, tiredness, and headache. Erythema, itch, heat sensation, edema and pain are also common symptoms of sunburn (UV dermatitis). Alterations have been observed in cell populations of the skin of patients suffering from so-called "screen dermatitis" similar to those observed in the skin damaged due to ultraviolet (UV) light or ionizing radiation. In "screen dermatitis" patients a much higher number of mast cells have been observed. It is known that UVB irradiation induces mast cell degranulation and release of TNF-alpha. The high number of mast cells present in the "screen dermatitis" patients and the possible release of specific substances, such as histamine, may explain their clinical symptoms of itch, pain, edema and erythema. The most remarkable change among cutaneous cells, after exposure with the above-mentioned irradiation sources, is the disappearance of the Langerhans' cells. This change has also been observed in "screen dermatitis" patients, again pointing to a common cellular and molecular basis. The results of this literature study demonstrate that highly similar changes exist in the skin of "screen dermatitis" patients, as regards the clinical manifestations as well as alterations in the cell populations, and in skin damaged by UV light or ionizing radiation. PMID- 9412816 TI - Chronic UVA (365-nm) irradiation induced scratching in hairless mice: dose-time dependency and the effect of ketanserin. AB - In a study on the dose-response relationship for longwave UVA (UVA1; 340-400 nm) carcinogenesis in hairless mice scratch marks appeared after months of daily exposure as an unwanted side effect. Tumor induction in the highest of the 4 tested dose groups (receiving a daily dose of 430 kJ/m2 of 365-nm radiation) could not be determined because extensive scarification occurred prior to the development of any tumors. The induction of scratch marks could be scored and quantified in all 4 dose groups tested. The UVA1 dose-dependencies for the induction of tumors and scratch marks were compared. We found that the induction of scratch marks depended mainly on the cumulative UVA1 exposure, whereas tumor induction showed a lesser dose-dependency. An attempt was made to prevent the apparent pruritogenic effect of UVA1 irradiation and to understand its mechanism. The influence of ketanserin, a serotonin/histamine antagonist, on the UVA1 induction of scratch marks was tested in groups of 8 mice daily irradiated with 430 kJ/m2. No difference was found between treated and untreated animals. Histological examination of skin biopsies from irradiated mice from the 430-kJ/m2 dose group from the UVA1 carcinogenic experiment, showed no changes in numbers of mast cells or other inflammatory features when compared to skin biopsies from unirradiated control mice. This indicated that UVA1-induced scratching is not mediated through mast cell release of serotonin and/or histamine. An adequate therapeutic treatment which can prevent UVA1-induced scratching would enable us to test tumor induction with UVA1 over a larger dose range, and may provide additional insight in how this radiation damages the skin. It remains conjectural whether there exists an analogous UVA-induced pruritus in human skin. PMID- 9412817 TI - Reverse iontophoresis: monitoring prostaglandin E2 associated with cutaneous inflammation in vivo. AB - In response to topical application of irritants, increased concentrations of prostaglandin E2 (PGE2) are found in human skin exudate and in cultured dermal fibroblasts. In this study, PGE2 generated in response to transdermal delivery of irritant drug compounds was monitored in hairless guinea pig (HGP) by a non invasive method, reverse iontophoresis. Reverse iontophoresis is the movement of molecules from the skin under the influence of an applied electric field. Irritant drug compounds were applied with iontophoresis (electrotransport), and reverse iontophoresis of PGE2 from skin was monitored by radioimmunoassay (RIA) after extraction from the delivery system. Chlorpromazine was used as a model drug irritant. When chlorpromazine and saline were applied over a range of current densities from 0 to 200 microA/cm2, visual scores of erythema and edema yielded a correlation with measured skin efflux of PGE2 (r = 0.86). Delivery of chlorpromazine resulted in greater efflux of PGE2 than delivery of non-irritant saline controls under the same delivery conditions. Five drug compounds, chloroquine, promazine, chlorpromazine, tetracaine, metoclopramide, and saline were applied to hairless guinea pig skin. The 6 agents were similarly rank ordered by visual erythema/edema scores and by PGE2 efflux, indicating that the quantity of PGE2 effluxed reflects the intensity of skin irritation. In contrast, vasoconstriction or vasodilation produced by the local delivery of vasoactive agents did not correlate with PGE2 skin efflux, indicating that this measurement is specific for an inflammatory response. In summary, PGE2 generated in response to transdermally applied drug irritants can be monitored non-invasively in vivo by reverse iontophoresis. PMID- 9412819 TI - Expression of c-fos in cultured human nevus cells: an increase over melanocytes and melanoma cells. AB - Nevus cells exhibit growth characteristics in culture which differentiate them from melanocytes and melanoma cells. We examined the expression of c-jun, c-fos and jun-B mRNA levels in cultures of different melanocytic cell types to determine if biologic differences among these cells was due to their level of proto-oncogene expression. Because cell growth and differentiation are also known to be affected by serum conditions, the expression of c-jun, c-fos and jun-B was examined under normal serum conditions and serum starved and repleted conditions which stimulates proto-oncogene expression. Expression of c-jun and jun-B was not significantly different among the cell types studied under normal serum conditions, or serum starved and refed conditions and c-fos was not detectable in any of the unstimulated cell types. In contrast, when the cells were serum starved and refed, the level of c-fos expression was uniformly increased (2-10 fold) in 3 different nevus cell lines. This increase was not seen in normal melanocyte cultures or 2 melanoma cell lines. With serum deprivation and repletion, c-fos was also elevated in 1 melanoma cell line. We conclude that the regulation of the proto-oncogene c-fos is different in nevus cells than in normal melanocytes, which may contribute to the different growth characteristics seen with nevus cells. PMID- 9412818 TI - Identification of the glycine-to-arginine substitution G2043R in type VII collagen in a family with dominant dystrophic epidermolysis bullosa from Hungary. AB - Epidermolysis bullosa (EB) represents a group of genodermatoses characterized by fragility and easy blistering of the skin. In the dystrophic forms of EB (DEB), blisters occur below the basement membrane, at the level of the anchoring fibrils. In the dominantly inherited forms (DDEB), the predominant type of mutation detected thus far is the substitution of a glycine residue which occurs within the collagenous domain of the molecule characterized by the repeating Gly X-Y amino acid sequence. In this study, we searched for mutations in DDEB in a family from Hungary, by PCR amplification of segments of COL7A1, followed by heteroduplex analysis. Examination of the PCR fragment corresponding to exon 73 revealed a heteroduplex in affected individuals from the family. Sequence analysis revealed a G-to-A transition at nucleotide 6127 in the triple-helical domain of COL7A1, which converted a glycine residue at amino acid position 2043 to an arginine. This report represents the second incidence of this mutation, G2043R, described first in a family with DDEB from Italy. PMID- 9412820 TI - The mode of action of prostaglandin (PG) I1 analog, SM-10906, on fibroblasts of hypertrophic scars is similar to PGE1 in its potential role of preventing scar formation. AB - The effects of prostaglandin (PG) I1 analog, SM-10906 (SM-6) and PGE1 on extracellular matrix formation and the release of cytokines by cultured normal human dermal fibroblasts (NDF) and hypertrophic scar fibroblasts (HSF) were compared in order to evaluate the clinical efficacy of PGs in preventing scar formation. In the present study, we measured type I collagen synthesis, collagenase activity, production of interleukin (IL)-6, IL-8, and transforming growth factor (TGF)-beta 1 and levels of adenosine 3,5-cyclic monophosphate (cAMP) in NDF and HSF cultured with or without PGs. The results demonstrated that HSF culture supernatants has a significantly higher level of type I collagen and TGF-beta 1 than those of NDF. However, the levels of collagenase activity and IL 8 in HSF were significantly lower in comparison to that of NDF. There was no substantial difference in IL-6 production between two types of culture cells. On the other hand, PGE1 and SM-6 significantly enhanced collagenase activity and raised the collagenase/type I collagen ratio in the HSF supernatants. In addition, both PGE1 and SM-6 increased production of TGF-beta 1, IL-8 and IL-6 and levels of cAMP in both cell types. However, they had no effect on the type I collagen synthesis of either types. These results suggest that, the stable PGI1 analog, SM-6, similarly acts as PGE1 in HSF by increasing the activity of collagenase. PMID- 9412821 TI - The 72-kDa and the 92-kDa gelatinases, but not their inhibitors TIMP-1 and TIMP 2, are expressed in early psoriatic lesions. AB - Psoriasis is histologically characterized by hyperkeratosis and papillomatosis with elongated vessels in the upper dermis. In order to evaluate the role of gelatinases in remodelling psoriatic skin in this study we examined the production of the 72-kDa (gelatinase A), 92-kDa collagenase (gelatinase B) and their tissue inhibitors TIMP-2 and TIMP-1. A total of 19 patients affected by different types of psoriasis were included in this study. An immunohistochemical study on cryosections was performed using antibodies to 72-kDa gelatinase, 92-kDa gelatinase, TIMP-1, TIMP-2, laminin, collagen types I, III, IV, VII. mRNA expression for gelatinases and their inhibitors were also analyzed by reverse transcriptase polymerase chain reaction (RT-PCR). In 14 of 19 patients there was a positivity in 92-kDa protein expression in keratinocytes. The 92-kDa gelatinase protein was also present in the upper dermis with prevalence around blood vessels. In 15 of 19 patients the 72-kDa was localized in the upper dermis, almost exclusively in the papillary dermis but absent in epidermis. TIMP-1 and TIMP-2 were both negative in all cases in immunoperoxidase and RT-PCR. Using RT PCR we show that the 72-kDa mRNA is expressed exclusively in the dermis, on the contrary the 92-kDa was present in epidermis and dermis. Type I, III, IV and VII collagens did not show any alteration or disruption. Overexpression and production of gelatinases without inhibitory effects suggest a role of these proteins in remodelling the psoriatic skin probably inducing the typical histological pattern of papillomatosis. PMID- 9412822 TI - Teaching nurses to think boldly in a time of chaos. AB - The world is undergoing revolutionary change, impacting on all sectors. Undaunted by the chaos and driven by growing patient needs, nurses are finding new ways of working, encompassing all that is good about nursing while concurrently breaking traditional barriers and expanding into new territory. PMID- 9412823 TI - A learning partnership in Ecuador. AB - Recognizing the need to provide community-based experiences for its undergraduates, the US' Spalding University of Louisville, Kentucky, teamed up with Catholic University in Quito, Ecuador, to fulfil this need and at the same time meet its School of Nursing's curriculum goal of developing concepts of caring and diversity. Below, how this partnership helped meet the objectives of a community health course in a diverse setting. PMID- 9412824 TI - Why lifelong learning? AB - The importance of professional development and what it means for nurses was described in a previous issue of International Nursing Review (March/April 1997). Below is a look at lifelong learning as reflected in continuing professional development for nurses, some international trends that influence health care, the international linkages taking place and the areas to be targeted for future professional development programmes. PMID- 9412825 TI - Continuing professional development: the European scene. PMID- 9412826 TI - The architecture of ICNP: time for outcomes--Part I. PMID- 9412827 TI - Ten-year experience with craniofacial implants: clinical and experimental results. AB - The choice of bone substitute to be used in the cranio-maxillofacial districts raises several problems relating to the biocompatibility and mechanical resistance of the material. Over the past 10 years a total of 245 cranio maxillofacial skeletal implants have been performed by the Department of Plastic Surgery at Niguarda Ca' Granda Hospital in Milan. The choice of material varied depending on the market availability of products and the experience acquired of both positive results and the onset of short-term and long-term complications. All patients were monitored with a minimum follow-up of 2 years and a maximum of 10 (mean 36 months). The following parameters were taken into account: complications linked to graft contamination, graft extrusion and decubitus, resorption times, degradable materials, resorption of underlying bone tissue and reactivity of the surrounding tissues. Studies on the cell cytohistotoxicity of materials used were performed in parallel by the cell culture laboratory at the Department of Plastic Surgery of Niguarda Hospital, using human keratinocyte and fibroblast cultures. PMID- 9412828 TI - New trends in the 3D management of CT data in plastic and reconstructive surgery. AB - Introduction of computer aided tomography in 1972 provided surgeons with multiple 2D maps which they themselves had to conceptualize mentally into a third dimension. The later advent of computerized summation of these data made it possible to display a perspective view of the third dimension on a TV monitor. CT, with the further analytical refinement afforded by software processing (interactive data presentation, contour detection and summation, hypothetical 3D data construction and interactive visualization) now provides the basic information that is needed for the fabrication of an individual model. Such models can be milled from polyurethane. More recently, laser-hardened acrylic resins have proved to be a useful alternative. Both systems are described and their advantages and disadvantages in the planning and performance of plastic and reconstructive surgical procedures discussed in the light of present knowledge. PMID- 9412829 TI - New trends in skin tumor surgery. AB - Over 8 years, 1700 patients were referred from the Mohs' Surgery and Cutaneous Laser Unit after Mohs micrographic skin tumor excision to the Division of Plastic and Reconstructive Surgery. Preoperative coordination between the two divisions was emphasised in wound preparation and timing of reconstruction for maximized patient convenience and outcome. Most repairs of facial and extremity defects were carried out under local anesthesia. Techniques of repair were selected based upon algorithmic priorities emphasizing simple techniques over complex ones. Direct closure, skin grafts and flaps were used. Preference for aesthetic subunit reconstruction of the face and the use of particular flap techniques including the O-to-S, O-to-T, V-to-Y island advancement, islandized nasolabial flap for alar reconstruction and the forehead flap for nasal dorsum and tip repair are illustrated. PMID- 9412830 TI - Eyelid reconstruction with irradiated human tarsal plate and aorta. AB - Reconstruction of posterior lamellar eyelid defects requires a tissue substitute that is either identical to the tissue lost (ex. surrounding or nearby tarsus) or donor tissue from another site that serves the same supportive role. Irradiated homologous tarsal plate and irradiated homologous aorta are potential posterior lamellar substitutes. Each provides a structural framework for the surrounding lid tissues to grow upon and are incorporated into the normal eyelid anatomy. Both the tarsal plate and aorta can be harvested, irradiated and stored in a refrigerator, ready to be utilized in those selected cases with severe tissue loss. They may also be utilized as a donor material in more routine lid reconstruction as an alternative. We discuss our experience with these materials. PMID- 9412831 TI - Microvascular surgery for the cancer patient. AB - The preview of reconstruction in the cancer patient has changed with the introduction of microsurgery and adjuvant therapy. Restoration of not only form and function but the quality of life must be considered when approaching these difficult and challenging patients. It is the purpose of this manuscript to present some background to microvascular reconstruction, and our approach to the cancer patient employed at The University of Texas MD. Anderson Cancer Center. By no means are the methods described absolute, but are meant to partially outline what has been our successful practice in some major areas in breast, head and neck and extremity reconstruction. With continuing efforts and the development of new innovative technologies, we can best achieve our goals of form, function and improved quality of life. PMID- 9412832 TI - The effect of heparinization on intra-abdominal infection and acute pulmonary failure. AB - Acute severe purulent peritonitis is still a very life-threatening disease condition. The primary cause of death is multiple organ failure (MOF) where acute respiratory distress syndrome (ARDS) is the initial trigger. Studies have shown that heparin has a cytoprotective effect and stimulates an increase in cardiopulmonary function systematically. METHODS: Twenty-one Sprauge Dawley rats were used as an animal model by puncturing the distal wall of intestine and ligating the appendix without interfering with the continuity of intestinal tract. In the formal experiments, 100 rats were divided into two groups and equal amounts of distilled water was given intraperitoneally. Total mortality and early mortality rate were recorded. Abdominal autopsies and blood gas analyses were performed and lung tissue samples were taken for light and electronic microscope analyses. RESULTS: The mortality rate was not statistically significant. But early death rate, the average survival times and the rate of abscess formation were significantly lower in the heparin treated group. The histological study of the experimental specimen showed that in the control group, the incidence of ARDS was higher and there was a more severe ARDS-like change, especially polymorphonuclear neutrophil leukocytes infiltration into alveolar and interstitial spaces. Blood gas analysis demonstrated the beneficial aspects of heparin administration. CONCLUSIONS: Heparin can increase the early survival rate and increase the survival time of rats with experimental acute severe purulent peritonitis, and proves to be beneficial in preventing infection induced lung injury during sepsis. We conclude that the effect of heparin on the survival rate is related to less pulmonary function deterioration, thereby preventing ARDS and the triggering of MOF. PMID- 9412834 TI - Refashioning of an aneurysmatic arterio-venous fistula by using the multifire GIA 60 surgical stapler. AB - We describe a technique for refashioning an aneurysmatic arterio-venous fistula by using the multifire GIA 60 surgical stapler. After obtaining proximal and distal control of the aneurysmatic vein each aneurysmal segment of the anterior wall of the vein is excised by applying the GIA 60 stapler. The layer of the staple-line is re-enforced with one layer of 6/0 prolene continuous suture. After completion of the procedure, the size of the vein is reduced by approximately 50%. The AVFs were successfully re-used for dialysis within four weeks postoperatively. PMID- 9412833 TI - The effect of corticosteroids and pentoxifylline in caustic esophageal burns. A prospective trial in rats. AB - BACKGROUND: Caustic Esophageal Burns (CEB) usually results in scatritial tissue and stricture formation. Management requires preventing the massive inflammatory process that ensues in its early phase and decreasing bacterial complications. METHODS: An animal model was created to investigate the effect of corticosteroids and pentoxifylline in CEB using 52 male Wistar rats. The injury was produced using an indwelling esophageal catheter through which 3N of 12% sodium hydroxide was infused. The rats were grouped as control, CEB, CEB and ceftazidime (CEB-C, 100 mg/kg/day im. bid. 10 days), CEB and ceftazidime plus dexamethasone (CEB-CD, 0.1 mg/kg/day im. bid. 4 weeks) and CEB and ceftazidime plus pentoxifylline (CEB CP, 50 mg/kg/day im. tid. 4 weeks). The groups were evaluated making use of esophagograms, hydroxyproline (OH-P) contents and histologic examination of the specimens 28 days after injury. RESULTS: No significant statistical differences were observed among the dexamethasone (CEB-CD), pentoxifylline (CEB-CP), antibiotics (CEB-C) and the untreated CEB groups. PMID- 9412836 TI - Efficacy of transgastrostomal endoscopic surgery (TGES) for early gastric cancer. AB - Early gastric cancer is now treated successfully by endoscopic mucosal resection (EMR). This technique, however, is not indicated when the tumoral lesion is located near the esophagogastric (EG)-junction, on the lesser curvature, or in the upper body or near the pylorus ring, and not indicated when the tumor size is greater than 20 mm. For these cases, we have developed what we term, "transgastrostomal endoscopic surgery" (TGES), using a Buess-type scope system. The aim of this study was to evaluate the efficacy of this technique in the treatment of those gastric cancers that could not be treated by EMR. In 4 patients selected for TGES, a Buess-type tube (external diameter: 40 mm) was inserted into the stomach through a temporary gastrostoma, and the whole operation was performed through the Buess-tube, using a video-camera for visualization. Using electrocautery scissors and forceps, complete resection of each lesion was performed, and the wound was closed by sutures. The average operation duration was 195 (130-240) minutes and the average blood loss was 59 (30-100) ml. The average size and margin of the resected specimens were 48 (30 59) and 13 (5-23) mm respectively. TGES is a substitutive, minimally invasive surgery to treat an early gastric cancer for which EMR would be difficult. This technique appeared to be simpler and easier than that of laparoscopic resection especially for a lesion on the posterior side of the stomach. PMID- 9412835 TI - Clinical presentations and predictive variables of thyroid microcarcinoma with distant metastasis. AB - BACKGROUND: Thyroid microcarcinoma is not an uncommon disorder. The purpose of this study is to analyze the clinical presentation and predictive factors for patients with thyroid microcarcinomas who have distant metastases. METHODS: We retrospectively reviewed and analyzed the clinical variables of 97 patients with thyroid microcarcinoma during the period from 1977 to 1995. The patients were divided into 2 groups representing patients with and without distant metastases. These data were analyzed by the Mann-Whitney U, chi 2 and Fisher's exact tests. RESULTS: Of the 97 patients with thyroid microcarcinomas, there were 6 (6.2%) cases (F/M = 5/1) with distant metastases. Among them, 4 cases were papillary carcinomas and 2 cases were follicular carcinomas. The parameters: age at diagnosis (P = 0.0137), one month postoperative serum thyroglobulin (Tg) level (P = 0.0215), cervical lymph node metastasis (P = 0.0097), and follicular cell type (P = 0.0079), were determined to be factors predictive for distant metastases by statistical analysis. There were no statistical differences between gender (P = 0.5781), postoperative 131I uptake (P = 0.1238), tumor size (P = 0.0571), preoperative thyroid function (P = 0.4425), fine-needle aspiration cytology (FNAC) (P = 0.9723), preoperative thyroid scan (P = 0.9765), and operative methods (P = 0.1060) between these two groups. CONCLUSIONS: Most thyroid microcarcinomas presented with relatively benign clinical courses, but patients with adverse predictive factors need more aggressive interventions to improve outcome. PMID- 9412837 TI - Serious complication of a blunt injury of the gluteal area. AB - The authors describe a rare case of the development of an extensive false aneurysm of the upper gluteal artery. The injury was caused by a blunt blow and was successfully operated on. PMID- 9412838 TI - Cytokine activation in patients undergoing open or laparoscopic cholecystectomy. AB - BACKGROUND: Laparoscopic surgery is estimated to produce a minor surgical injury in comparison with open and laparoscopic cholecystectomies. Studies in the past compare almost data of the first hours until day two. However, the surgical injury and the wound healing metabolism has to be detected. METHODS: A prospective study was designed to investigate cytokine responses after surgical injury. Twenty-three patients with symptomatic cholelithiasis were admitted to the study. Eleven patients underwent conventional (open) and twelve patients laparoscopic cholecystectomy. Circulating concentrations of cytokines-including Interleukin-6 (IL-6), Tumor necrosis factor-alpha (TNF-alpha) and neopterin, were compared between both groups. In addition, association of the cytokines with clinical parameters including leucocytes, urea, fever and C-reactive protein (CRP) were assessed. We are using ELISA-test of Medgenix GmbH, Ratingen, Germany and BRAHMS Diagnostica, Berlin, Germany. RESULTS: Enhanced cytokine responses were observed in the conventional group compared to the laparoscopic group. On day 3 after operation, the second increase in cytokine levels (but smaller than the first increase) were observed in both groups. In the conventional group, a slightly high correlation ratio between the urea and cytokine levels was found. However, only neopterin and urea association on postoperative day 3 (r = 0.65) was significant (p < 0.05). There was no significant association between CRP and cytokine levels in both groups. CONCLUSIONS: Cytokine response after cholecystectomy demonstrates the lesser degree of surgical injury in the laparoscopic group, however, TNF-alpha demonstrates on day 4 a similar increase in both groups. This is a new result of studies working in this field. In conclusion, the benefit of laparoscopic surgery results only in the minimal access to the abdominal cavity, the wound healing metabolism is at last the same in both groups. PMID- 9412839 TI - Surgical treatment of paragangliomas of the neck. AB - Of a total of 5,700 surgical procedures on the neck performed at our Institution between 1984 and 1995, 13 operations (0.22%) have been done on 11 patients with 16 cervical paragangliomas. A woman underwent resection of synchronous bilateral carotid body tumours and of an intravagal paraganglioma. Ten years later, after preoperative angiographic embolization, she underwent resection of a paraganglioma of the left hypoglossal nerve. Her sister, at age 21, underwent resection of a carotid body tumour and, respectively 19 and 20 years later, of a right and left intravagal paraganglioma. An interposition graft for replacement of the carotid bifurcation was necessary in one patient only. During resection of a left carotid body tumour, acute hypotension occurred resulting in an ischemic lesion of the right temporal lobe. Postoperatively, she also complained of respiratory distress that responded to medical therapy with difficulty. The related neurologic symptoms completely resolved three months after surgery. The operation for a paraganglioma of the left hypoglossal nerve resulted in a temporary motor deficit of the tongue and in permanent considerable difficulty in swallowing. Unilateral recurrent nerve palsy occurred in two patients. No patients during the postoperative follow-up showed signs of local recurrence or metastatic disease. In conclusion, surgery is an effective treatment for cervical paraganglioma, but because of the high surgical complication rate, an experienced and skilled surgeon is called on to optimize outcome. An adequate perioperative care is advisable. PMID- 9412840 TI - Activity and localization of cathepsin B, D and G in aortic aneurysm. AB - The activities of cathepsin B, D and G, and their distribution in the walls of normal aorta and those of aneurysms of abdominal aorta were studied. The immunohistochemical reaction for these cathepsins was positive in the aneurysm wall both in the cells and in the extracellular matrix. It was more apparent than in normal aortas. The activities of cathepsin B, D and G in aneurysm were 1.5 times higher than in normal aortas. It may be suspected that besides the action of collagenases and elastases the cathepsins participate in an enhanced degradation of collagen and elastin fibers of aneurysm wall. PMID- 9412841 TI - Preoperative methylene blue staining of galactographically suspicious breast lesions. AB - Microdochectomy is the standard treatment of galactographically suspicious breast lesions. Precise preoperative marking of the suspicious duct and intraductal lesions facilitates selective minimal-volume microdochectomy. Methylene blue dye staining fulfills this criterion. A retrospective review of our experience of preoperative methylene blue staining in 30 patients with unilateral spontaneous nonlactiferous single duct nipple discharge operated on during 1986-1995 in the Oulu University Hospital for galactographically suspicious breast lesions. Galactography was successful in 29 out of 30 (93.3%) cases. Preoperative methylene blue staining was attempted in all cases on the day of surgery and it was successful in 22 (73.3%) cases making subsequent selective minimal-volume microdochectomy easy to perform. The failure of methylene blue staining led to quadrantectomy in 4 cases and smaller breast resections in the remaining 4 cases. Preoperative methylene blue dye staining crucially facilitates selective minimal volume microdochectomy. An interval between primary galactography and later methylene blue staining leads to failures in approximately one quarter of the cases. A higher success rate would necessitate scheduling the microdochectomy on the same day as the primary galactography (and the subsequent methylene blue staining in suspicious cases). PMID- 9412842 TI - Videoassisted transhiatal esophagectomy for cancer. AB - Blunt transhiatal esophagectomy is largely performed in selected cases of esophageal cancer according to the experience of Mark Orringer. We have recently performed eleven consecutive videolaparoscopy assisted transhiatal esophagectomies in order to help esophageal dissection and to avoid injuries to mediastinal structures. In our experience the routine use of laparoscopic assistance during transhiatal esophageal dissection improves the safety of this technique and lowers postoperative complications. The results of neoadjuvant treatments (radio-chemotherapy) recently reported emphasize the role of transhiatal esophagectomy for cancer. PMID- 9412843 TI - MRI and ultrasonographic findings in the investigation of lymphedema and lipedema. AB - METHODS: Twenty-four healthy subjects and 16 patients with lymphedema and lipedema were studied with MRI and ultratomography. RESULTS: In chronic lymphedema, ultrasonography revealed a statistically significant increase of the subcutaneous fat without difference in skin thickness as compared to the healthy subjects. MRI revealed in lymphedema a statistically significant increase of skin thickness + subcutaneous tissue + muscular mass (p = 0.048); in lipedema, a statistically significant increase of skin thickness and subcutaneous tissue (p < 0.0001) as compared to the healthy controls. CONCLUSIONS: MRI offers strong qualitative and quantitative parameters in the diagnosis of lymphedema and lipolymphedema, while ultrasonography is expected to improve its diagnostic efficiency with the aid of high frequency echo with more sophisticated resolution apparatus. Age, weight and height of the patient as well as duration of the disease do not seem to affect the above-mentioned parameters. PMID- 9412844 TI - Total thyroidectomy or lobectomy in benign nodular disease of the thyroid: changing trends in surgery. AB - METHODS: Over a 23-month period (January 1994-December 1995) in the era of fine needle aspiration (FNA), 344 thyroid surgery operations were performed for benign diseases of the thyroid. Of these 55 total thyroidectomies or lobectomies were evaluated. Mean age was 43.6 + 9.7 and the female/male ratio was 47/8 (5.8). All cases were operated on with the consensus of the surgery + endocrinology + pathology council according to a protocol based on FNA, thyroid function tests, scintigraphy and ultrasound. Suspected FNA or suspected nodules during the surgery were verified by frozen section also. Of these 55 benign nodules, 7 (12.7%) had total bilateral, 48 (87.3%) unilateral lobectomies. RESULTS: Postoperatively 3 cases (5.8%) of seroma, one transient hypoparathyroidism (1.8%) and one unilateral vocal cord paralysis (1.8%) were seen as complications. CONCLUSIONS: In our center, FNA cytology has been a routine procedure since 1992. Surgery for benign thyroid disease has been reduced 50% since than. This study was started after two years of the FNA procedure. Resident nodules are the most common cause of recurrence in nodular thyroid disease, so some cases need radical surgery when it is decided to operate. Morbidity of surgery for recurrent disease is unacceptably high and is comparable to lifelong supplement therapy. PMID- 9412846 TI - Surgical management of patients with radiation enteritis. PMID- 9412845 TI - Spontaneous perforations of the small intestine. AB - BACKGROUND AND METHODS: The present study describes the procedures used by the authors in the management of 34 patients with spontaneous perforation of the small intestine. RESULTS: Only one (2.9%) of the patients had the perforation cause diagnosed before laparotomy; 27 (80%) cases showed ileal perforative lesions while seven (20%) had jejunal lesions; 31 (91.1%) patients presented single perforations and three (8.8%) had multiple ones. Intestinal resection followed by anastomosis or ileostomy and colostomy, was carried out in 21 (61.7%) cases, and 13 (38.2%) patients were submitted to exeresis with edge restoration and lesion suture. The cause of perforation could be identified in 29 (86.3%) cases while in five (14.7%) patients the cause was considered idiopathic. Eighteen (53%) patients recovered from surgery and were discharged; there were 16 (47%) deaths resulting from a number of complications. CONCLUSIONS: Since the prognosis regarding this disease depends on the peritoneal infection severity level, the patient's organic resistance, and most of all, the time interval spent until the treatment is initiated, the authors emphasize the need to have a laparotomy performed as early as possible considering that this procedure provides the best chances of survival and health recovery. PMID- 9412847 TI - New clinical signs. PMID- 9412848 TI - Parathyroid cysts. PMID- 9412849 TI - Guidelines for the therapeutic use of botulinum toxin in movement disorders. Italian Study Group for Movement Disorders, Italian Society of Neurology. AB - Since its introduction in the early '80s the use of botulinum toxin has improved the quality of life of the patients affected by movement disorders. Toxin's neuromuscular blocking action allows a symptomatic treatment of those clinical conditions characterised by excessive muscular activity. Although the dosages used are safe and the side-effects are reversible, a correct use of botulinum toxin depends on the knowledge of its clinical pharmacology and of the anatomy of the body segments to be injected. In addition, the treatment of more complex conditions, i.e. laringeal dystonia, imposes an inter-disciplinary approach and specialised injection techniques. In this review, the Italian Study Group on Movement Disorders presents the consensus guidelines for the therapeutic use of botulinum toxin in movement disorders. The main toxin types, their use and administration modalities, and the training guidelines will be presented. PMID- 9412850 TI - Autosomal recessive distal muscular dystrophy. AB - The "distal myopathies" include autosomal dominant, autosomal recessive, and sporadic disorders. Two of the recessive disorders are considered to be definitive entities: Miyoshi's myopathy, which has an early adult onset and first involves the calf muscles, and distal myopathy with rimmed vacuoles. We here describe the cases of two sisters and compare them with previously reported cases. The disorder in our patients is characterised by: a) autosomal recessive inheritance; b) onset in early adult life; c) initial involvement of the tibialis anterior and peroneal muscles; d) subsequent involvement of the calf muscles spreading to the proximal muscles of the legs and, later, the arms; e) a moderately disabling evolution over a period of 10-12 years; f) marked and stably high serum levels of CK and other enzymes; g) EMG evidence of myopathic damage, with fibrillation at rest; and h) a histological picture of dystrophic myopathy, with atrophy of mainly type 2 fibres. We think that this syndrome is different from the two forms of autosomal recessive distal myopathy mentioned above. PMID- 9412851 TI - Searching for migraine genes: exclusion of 290 cM out of the whole human genome. AB - A linkage and association analysis was made on 14 Italian families with recurrent migraine. We analyzed five chromosomal regions surrounding the candidate genes 5HT1D (1p36.3-34.3), 5HT1B (6q13), 5HT2A (13q14-21), 5HT transporter (17q11.2 12), CACNLB1 (17q11.2-22) and FHM (19p13), using 29 DNA polymorphic markers. All two-point lod scores were negative, and the chi 2 sib-pair analyses were not significant, thus indicating the probable exclusion of these regions as sites of migraine genes in our population. PMID- 9412852 TI - Periodic syndrome and migraine in children and adolescents. AB - Many reports in the literature seem to confirm the hypothesis that the symptoms of periodic syndrome are precursors or the equivalent of migraine: the aim of this study was to assess the prevalence of periodic syndrome in a group of children and adolescents suffering from migraine in comparison with that observed in various control groups. We considered seven symptoms: recurrent vomiting and abdominal pain, migrating limb pain, vertigo, recurrent hyperthermia with no visible cause, sleep disturbances and eating disorders. The study involved 171 children divided into four groups; 42 migraineurs; 37 subjects with chronic nervous pathologies but no psychosomatic symptoms; 46 subjects with a known psychosomatic disease, and 46 healthy subjects. The prevalence of the symptoms in the different control groups was different, although the pattern was more similar in the migraineurs and psychosomatic patients than in the other control groups. The development continuum of the syndrome may support the view that periodic syndrome is predictive of the subsequent development of a psychosomatic pathology. PMID- 9412853 TI - Pudendal nerve somatosensory evoked potentials in probable multiple sclerosis. AB - Bladder dysfunctions are often observed in patients with multiple sclerosis (MS). In order to evaluate their sensitivity in detecting abnormalities in bladder central control pathways, pudendal nerve somatosensory evoked potentials (pSEPs) were recorded in 16 patients with clinically probable MS: six were affected by retention or urge incontinence, and ten were asymptomatic. Conventional visual, auditory and somatosensory evoked potentials were also recorded, and all of the patients underwent a urodynamic examination. Prolonged latency or the absence of pSEP cortical responses was found in eight of the ten asymptomatic patients, and in all of the symptomatic cases (87.5%). The urodynamic evaluation revealed abnormalities in 12 patients (75%). Our findings seem to indicate an early and frequent involvement of bladder control pathways in MS patients, as well as a high rate of subclinical disorders. PMID- 9412854 TI - Adie's tonic pupil as a manifestation of Sjogren's syndrome. AB - We here report two cases of Adie's tonic pupil, associated with clinical sensory polyneuropathy and Sjogren's syndrome, in one of whom it actually heralded the onset of the syndrome. Electrophysiology studies indicated absent H reflexes but normal peripheral nerve conductions, thus suggesting an involvement of the dorsal roots or spinal ganglion that would be in line with previously published reports of dorsal ganglionitis as the primary neuropathological lesion in Sjogren's syndrome. We suggest that all cases of tonic pupils should be screened for polyneuropathy and Sjogren's syndrome. PMID- 9412855 TI - Low back pain due to hypertrophic roots as presenting symptom of CIDP. AB - Attention has recently been drawn to chronic inflammatory demyelinating polyneuropathy (CIDP) with symptomatic nerve root hypertrophy. A 31-year-old woman had fluctuating and worsening low back pain. Absent tendon jerks and a slight weakness of the hand interossei muscles suggested a diffuse neuropathy. The electrophysiological and histological findings were diagnostic for CIDP. Lumbar spine MRI showed marked nerve root enlargement with gadolinium enhancement. This case widens the range of the clinical presentations of CIDP. Further studies are warranted to ascertain whether cauda equina gadolinium enhancement may be a useful tool in the diagnosis of CIDP and a marker of disease activity for monitoring response to therapy. PMID- 9412856 TI - Dystonia as an isolated symptom of multiple sclerosis? AB - Sustained dystonia has seldom been reported during the course of multiple sclerosis (MS) [5-8, 10], and has been described as the first manifestation of the disease in only three cases [1,3]. We describe a patient with a diagnosis of laboratory-supported, defined MS in which sustained dystonia was the only neurological symptom. PMID- 9412857 TI - Pain due to epidural tumor in cancer patients. Report of two cases and differential diagnosis. AB - The cases of two patients with inguinal pain as the only symptom of a T12 metastatic lesion is reported. The patterns of pain referrals from tumor lesions to the spine, epidural space, and spinal cord are reviewed. Focal back pain and pain reported in a distal distribution can both be associated with epidural or cord disease. The differential diagnosis of back pain in patients with cancer can be difficult but may be crucial in differentiating important neurological complications of systemic neoplasms. PMID- 9412858 TI - Malignant ischemic stroke in the carotid district. PMID- 9412859 TI - Perforin-secreting killer cell infiltration in the aortic tissue of patients with atherosclerotic aortic aneurysm. AB - Cell-mediated immunity has been implicated in the pathogenesis of vascular cell injury in patients with atherosclerotic aortic aneurysms. To clarify the immunologic mechanisms involved, we examined the expression of a cytolytic factor, perforin, in infiltrating cells from aortic tissue samples taken from 6 patients with atherosclerotic aortic aneurysms. Immunohistochemical studies showed that the infiltrating cells consisted mainly of macrophages, natural killer (NK) cells, cytotoxic T lymphocytes (CTLs), and T helper cells, and that perforin was expressed in NK cells and CTLs. Immunoelectron microscopic studies demonstrated that the infiltrating cells released massive amounts of perforin directly on to the surface of arterial vascular cells. These findings provide the first direct evidence that some of the infiltrating cells in the aortic tissue consist of killer cells, and strongly suggest that these killer cells, especially NK cells and CTLs, may play a critical role in the vascular cell injury caused by atherosclerotic aortic aneurysm by releasing perforin. PMID- 9412860 TI - Correlations between recruitable coronary collateral flow velocities, distal occlusion pressure, and electrocardiographic changes in patients undergoing angioplasty. AB - Direct assessment of coronary collateral flow has been difficult in humans. The goal of this study was to correlate the magnitudes and waveform characteristics of recruitable coronary collateral flow velocity measured with Doppler guidewire with other hemodynamic and functional indexes of collateral circulation in patients during angioplasty. Twenty-six patients [age 60 +/- 10 (SD) years] were studied for measurements of flow velocity at the distal segment of the dilated vessel. Collateral flow signals were demonstrated in 18 patients (69%) during balloon inflation. There was a weak yet significant positive correlation between the magnitude of collateral flow velocity and distal occlusion pressure (p < 0.01). The sum of ischemic ST elevation on the 12-lead electrocardiogram was inversely correlated with distal occlusion pressure, but not with the magnitude of collateral flow velocity. Subgroup analysis between patients with (n = 18) and without (n = 8) ST elevation during balloon inflation revealed higher collateral flow velocity signals (p < 0.0001) and greater systolic components of collateral flow in the latter group (p < 0.0001). Thus, functionally significant coronary collaterals showed greater velocity signals and systolic predominance in flow pattern. The functional capacity of human coronary collaterals may be semiquantitated using the Doppler guidewire technique. PMID- 9412861 TI - The rise time of the monophasic action potential--a new index of local use dependent conductivity by sodium channel blockers in human myocardium. AB - The kinetics of global use-dependent conduction slowing produced by sodium channel blockers in the human heart, estimated as a change in the QRS width, are known to be similar to those of use-dependent block of the maximum rate of depolarization in in vitro studies. However, the kinetics of the regional use dependent decrease in conductivity have not been investigated. We examined whether the rise time of the monophasic action potential would be clinically useful as a marker of the local use-dependent decrease in conductivity by sodium channel blockers. In 12 patients without organic heart disease, monophasic action potentials (MAPs) were recorded at the right ventricular endocardium using a contact electrode before and after the administration of disopyramide (n = 6, 2 mg/kg, i.v.) or pilsicainide (class Ic agents, n = 4, 1 mg/kg, i.v., and n = 2, 150 mg, po) while the stimulus frequency was abruptly increased from 100/min to 150/min. The rise time, defined as the interval from the pacing pulse to the first peak deflection of the monophasic action potential, and the ORS width were measured simultaneously. In the absence of the sodium channel blockers, the abrupt increase in heart rate did not alter the QRS width or the rise time. In the presence of the agents, both variables were lengthened exponentially. The rate constants of onset changes in the QRS width and the rise time were 2.1 +/- 0.5 beats and 2.1 +/- 0.4 beats after the administration of disopyramide, and 7.5 +/- 3.0 beats and 8.2 +/- 4.0 beats after pilsicainide, respectively. The rate constant of the rise time was closely correlated with that of the QRS width. The present results are very closely comparable with the onset rate constants of use dependent block of the maximum rate of depolarization in in vitro studies. These results suggest that (1) the rise time is a good indicator of local use-dependent decrease in conductivity by sodium channel blockers in human hearts and (2) the local use-dependent decrease in conductivity has kinetics similar to those of use dependent sodium channel blocks. PMID- 9412862 TI - Electrophysiologic changes in arrhythmogenic substrate following the maze procedure in patients with lone and paroxysmal atrial fibrillation. AB - Although the Maze procedure is highly successful in restoring sinus rhythm in patients with atrial fibrillation (AF), the electrophysiologic changes that occur after the Maze procedure and its mechanism of action are still unclear. The aims of the present study were to examine the electrophysiologic changes that occur in the arrhythmogenic substrate after the Maze procedure and to evaluate the mechanism by which it prevents lone and paroxysmal AF. The modified Maze procedure was performed in 6 patients (6 men, mean age 47 +/- 7 years) with lone and paroxysmal AF. Electrophysiologic studies were performed before and 35 +/- 8 days after the Maze procedure. Atrial mapping during sinus rhythm revealed that atrial activation propagated smoothly in a concentric circle from the sinus node to other regions in the right atrium and into the coronary sinus before the Maze procedure. Following the Maze procedure, atrial activation propagated selectively through routes produced by incisions or cryoablations in all 6 patients. In addition, double and triple potentials were recorded in regions where conduction blocks were created by the Maze procedure. Neither sinus node function nor AV conduction was changed following the Maze procedure. The Maze procedure did not affect the atrial effective refractory period or the zone of fragmented atrial activity, although the conduction delay zone was increased significantly (p < 0.05). AF was inducible in all 6 patients before the Maze procedure, whereas it was not inducible in any patients after the Maze procedure. The Maze procedure is effective in preventing AF without affecting sinus node function and AV conduction. Intra-atrial conduction block produced by incisions or cryoablations contributes to the prevention of AF. PMID- 9412863 TI - Coil occlusion for patent ductus arteriosus in Japan. AB - We surveyed Japanese experience of coil occlusion of patent ductus arteriosus up to 30 September 1996 by sending questionnaires to 175 hospitals. Thirty-four hospitals reported outcome data for 231 procedures in 218 patients. Successful implantation was achieved in 94% and acute complete closure of the ductus occurred in 71% of those in whom implantation of the coil was successful. Of the latter, 83% reported late complete closure. When those patients who underwent reocclusion for residual shunt are included, 89% attained complete closure. No life-threatening complications have occurred so far. Late reopening was reported in 3 cases. Although the angiographic type of ductus was significantly related to successful implantation (p < 0.01), there was no significant correlation with complete occlusion. Ductuses with a minimum diameter greater than 3 mm had a decreased chance of successful implantation, whereas those less than 2 mm had a greater incidence of complete closure. PMID- 9412864 TI - Sudden cardiac death in Japanese people aged 20-60 years--an autopsy study of 133 cases. AB - In order to elucidate the principal cause and actual circumstances of sudden cardiac death in Japan, especially among people in the prime of life, we investigated 133 out of 161 autopsied patients (82.6%) (106 men and 27 women, mean age 47.5 years). Coronary artery disease (CAD) was the most frequently detected disorder (50 cases, 37.6%), and included 15 cases of acute myocardial infarction (AMI) (11.3%). We found that CAD was less frequent among younger patients than in Western countries: 10.0% in subjects in their twenties and 22.2% among subjects in their thirties. The left anterior descending artery (LAD) was the vessel most often affected by infarction (47.0%), but the proportion of LAD lesions was not different from that in AMI patients who were survived for least 1 day after the attack. In conclusion, CAD was infrequent among patients aged 20-39 years in comparison with Western countries and LAD was the most commonly affected vessel, but the proportion was not different from that found among AMI patients who survived for at least 1 day after the attack. PMID- 9412865 TI - Internal mammary hypoperfusion syndrome--diagnosis and treatment. AB - The use of the internal mammary artery (IMA) in coronary artery bypass surgery has increased substantially over the past 20 years, being at present the conduit of choice for most patients. Complications associated with its use occur occasionally and include life-threatening postoperative ischemia or the revascularized myocardium. We reviewed the records of 1,971 consecutive patients who underwent coronary artery bypass grafting over a 5-year period. All operations included an IMA graft to the left anterior descending coronary artery. Twenty-eight of these patients (1.4%) underwent additional placement of a vein graft on the same region as a salvage maneuver for suspected hypoperfusion as a result of IMA failure. All 28 patients showed life-threatening hemodynamic compromise. Twenty-two of the 28 patients (79%) survived. This was the result of immediate surgical correction, which reversed their hemodynamic instability. IMA hypoperfusion was found more frequently in reoperations and in women and diabetic patients. This syndrome is the result of an imbalance between IMA flow and myocardial demand, causing sudden and unexpected myocardial failure. Its detection and expeditious treatment can successfully modify a serious and potentially lethal clinical situation. PMID- 9412866 TI - Effects of volume loading on pulmonary venous flow and its relation to left atrial functions. AB - Although pulmonary venous (PV) flow is closely related to left atrial (LA) pressure dynamics, few investigators have discussed it in relation to LA functions, i.e., reservoir, conduit, and booster pump functions. We examined changes in PV flow rate, LA dimension, and left ventricular filling volume in 11 dogs, and assessed the effects of multistaged volume loading on PV flow and LA functions. Systolic PV flow rate (S) increased significantly and reached a plateau, reflecting a limited LA reservoir function. Diastolic PV flow rate (D) increased significantly with an increase in LA pressure. S/D ratio increased non significantly from 0.87 +/- 0.07 before volume loading to 0.96 +/- 0.08 until S reached a plateau and then decreased to 0.76 +/- 0.08 (p < 0.05) because of a significant increase in D without an increase in S at the higher stages of volume loading. During atrial contraction, increases in LA active shortening and left ventricular filling volume were limited, indicating a limited LA forward ejection. The difference between PV flow rate just before and at the end of atrial contraction increased and correlated positively with left ventricular end diastolic pressure (r = 0.57, p < 0.01). PV flow varies according to the degree of volume loading and reflects LA functions, which exhibit limited increases in response to volume loading. PMID- 9412867 TI - Protective effect of amiloride against reperfusion damage as evidenced by inhibition of accumulation of free fatty acids in working rat hearts. AB - To examine whether amiloride protects against ischemia-induced or reperfusion induced damage to the heart, mechanical and metabolic studies were performed in the isolated, working rat heart. Ischemia decreased both mechanical function and the tissue levels of high-energy phosphates and increased the tissue levels of free fatty acids (FFAs). Reperfusion restored the levels of high-energy phosphates but further increased FFA accumulation. For this reason, accumulation of FFAs was used as an indicator of both ischemia-induced and reperfusion-induced damage. Drugs were added to the perfusion solution 5 min before ischemia until the end of ischemia (pre) or until 10 min after reperfusion (pre + post). Diltiazem (1 or 5 mumol/L pre) decreased the mechanical function of the non ischemic heart and attenuated both ischemia-induced and reperfusion-induced accumulation of FFAs. Amiloride (50 mumol/L pre) did not affect the mechanical function of the non-ischemic heart or attenuate ischemia-induced or reperfusion induced FFA accumulation effectively. However, amiloride (50 mumol/L pre + post) did markedly attenuate the reperfusion-induced accumulation of FFAs. In conclusion, diltiazem attenuates both ischemia-induced and reperfusion-induced myocardial damage, probably through its energy-sparing effect as a result of a decrease in mechanical function before ischemia. In contrast, amiloride attenuates only the reperfusion-induced myocardial damage through mechanisms other than the energy-sparing effect. PMID- 9412869 TI - Acutely severe myocarditis successfully treated by percutaneous cardiopulmonary support applied by a newly developed heparin-binding oxygenator and circuits. AB - The feasibility of using the heparin-bound percutaneous cardiopulmonary support system (PCPS) for prolonged extracorporeal circulation in patients with acute severe myocarditis is demonstrated. The case histories of 2 patients with cardiogenic shock caused by acute myocarditis are presented; both were successfully treated with long-term PCPS using a newly developed heparin-binding oxygenator and circuits without changing the oxygenator. The courses of both patients remain uneventful more than 12 months after discharge. We also discuss the clinical aspects of using heparin-bound PCPS in patients with acute severe myocarditis. PMID- 9412868 TI - Regulation of bradykinin-stimulated cation entry into endothelial cells by tyrosine kinase. AB - In endothelial cells, bradykinin stimulates the release of intracellular Ca2+, which is followed by the entry of extracellular Ca2+ into the cells. However, the mechanism underlying the Ca2+ entry is not well understood. To investigate the possible implication of tyrosine kinases in bradykinin-mediated Ca2+ signaling in endothelial cells, cultured porcine aortic endothelial cells were loaded with fura-2/AM, and Mn2+ influx into the cells was determined by the quenching of fluorescence intensity of fura-2 at 360 nm excitation. The tyrosine kinase inhibitors genistein and herbimycin A attenuated not only Ca2+ influx but also Mn2+ influx from the extracellular space without affecting the release of Ca2+ from internal stores in bradykinin-treated cells. In contrast to tyrosine kinase inhibitors, the tyrosine phosphatase inhibitor vanadate stimulated Ca2+ influx as well as Mn2+ influx. On the other hand, both an inactive analog of genistein, daidzein, and an inhibitor of diacylglycerol (DAG) kinase, ethylene glycol dioctanoate, were without effect on [Ca2+]i following the stimulation of agonist. These findings suggest that tyrosine kinase is involved in the regulation of cation influx in endothelial cells. PMID- 9412871 TI - [Significant role of new cephem antibiotics: Focused on cefozopran]. PMID- 9412870 TI - Magnetic resonance imaging in diagnosis of right coronary arteriovenous fistula- a case report. AB - We describe a patient with a right coronary arteriovenous fistula in whom magnetic resonance imaging (MRI) was useful in establishing the diagnosis. In a 36-year-old woman, T1 spin echo MRI demonstrated a massively dilated coronary arteriovenous fistula connecting the right coronary artery to the right atrium. The cine field echo technique showed a continuous shunt flow within the fistula as documented by the flow void throughout the cardiac cycle. These findings were confirmed by cardiac catheterization. We conclude that MRI is useful not only in detecting a coronary arteriovenous fistula but also in identifying its origin and the drainage site. PMID- 9412872 TI - [Initial treatment at an outbreak of E. coli O-157:H7 infection: especially with respect to therapy in the emergency]. AB - An outbreak of O-157:H7 diarrheal illnesses occurred in junior schools of Sakai city, Osaka prefecture, in last July, 1996. At the beginning of the outbreak, many patients rushed to outpatient clinics. From the practical experiences, we examined the necessity of fluid therapy in patients regarding their initial clinical features. The risk factors for development of HUS were noted as presence of fever, WBC counts of more than 10,000/microliter and more than 1.0 mg/dl of CRP. During the prodoromal illness, administration with available antimicrobial agents would be advisable for high risk patients, while it would yet be remain to be further investigated. The majority of the patients with clinical manifestations showed neither signs for dehydration nor electrolyte abnormalities based on the blood examination data and biochemical analysis of the serum. Therefore, the fluid therapy did not appear necessary for majority of patients except a few high risk patients, when outpatient clinics were crowded with emergency patients. PMID- 9412873 TI - [The drug susceptibility pattern of the presumed etiologic agents of infectious enteritis including verotoxin-producing Escherichia coli O-157]. AB - The drug susceptibility patterns were investigated for verotoxin-producing Escherichia coli (VTEC) including O-157, Salmonella spp., Vibrio parahaemolyticus and Campylobacter jejuni subsp. jejuni that were obtained in and after July 1996. The results are summarized as follows; 1. We found highly resistant strains of VTEC to tetracycline (TC) and ampicillin (ABPC). Minimum inhibitory concentrations (MIC) of some of the drugs against VTEC in an aerobic condition were significantly different from those in an anaerobic condition. For example, aerobic.anaerobic MIC ranges of the drugs tested were as follows: chroramphenicol (CP): 1.56-3.13 micrograms/ml.0.78-1.56 micrograms/ml, TC: 1.56-> 100 micrograms/ml.0.78-> 100 micrograms/ml, minocycline (MINO): 1.56-12.5 micrograms/ml.0.78-3.13 micrograms/ml, kanamycin (KM): 3.13-6.25 micrograms/ml.25 100 micrograms/ml, fosfomycin (FOM): 3.13-25 micrograms/ml.0.78-6.25 micrograms/ml, norfloxacin (NFLX): < or = 0.025-0.2 microgram/ml.< or = 0.025-0.2 microgram/ml, ABPC: 1.56-> 100 micrograms/ml.0.78-> 100 micrograms/ml and cefaclor (CCL): 1.56-25 micrograms/ml.56-12.5 micrograms/ml. MICs of CP and tetracyclines (TCs) in an anaerobic condition were lower by twofold and the MIC of FOM was lower by fourfold, but the variabilities of MIC-ranges of NFLX, ABPC and CCL were small. The MIC of KM was high. 2. We observed that some of Salmonella spp. were highly resistant to CP, TCs and MINO, and some were moderately resistant to NFLX. 3. The detection frequency of TC-resistant strains and NFLX-insensitive or resistant strains were high among C. jejuni subsp. jejuni. 4. Strains of V. parahaemolyticus and C. jejuni subsp. jejuni were mostly resistant to ABPC and CCL, MICs of which were in high ranges. 5. Fecal concentrations in MINO, KM, FOM and NFLX reported in literatures are high enough to have some antimicrobial activities, leed dose of ABPC and CCL are quite low. PMID- 9412875 TI - Transfusion algorithms: more than just alphabet soup? PMID- 9412874 TI - [Comparative antimicrobial activity of RP 59,500 (quinupristin-dalfopristin), the first semisynthetic injectable streptgramin, against gram-positive cocci and other recent clinical pathogens]. AB - RP 59,500 (Quinupristin-Dalfopristin) is the first semisynthetic injectable streptogramin antimicrobial agent, which is a combination of quinupristin and dalfopristin in a 30:70 ratio. The components of RP 59,500 act synergically to provide bactericidal activity through action at different sites on bacterial ribosomes. In the present study, the antimicrobial activity of RP 59,500 was compared with those of four macrolides (erythromycin, clarithromycin, azithromycin, roxithromycin). Susceptibility testing was carried out by microdilution method on 303 strains of 10 species, especially antibiotic resistant Gram-positive cocci. RP 59,500 was active against a wide range of Gram positive cocci including methicillin-resistant Staphylococci and penicillin resistant Streptococcus pneumoniae. The MICs90 of RP 59,500 against methicillin resistant Staphylococcus aureus (MRSA) and Staphylococcus epidermidis were both 0.25 microgram/ml, although those of four macrolides were higher than 32 micrograms/ml. The MICs90 of RP 59,500 against penicillin-sensitive, intermediate and -resistant S. pneumoniae were all 0.5 microgram/ml, although those of four macrolides against penicillin-resistant S. pneumoniae were higher than 32 micrograms/ml. RP 59,500 also exhibited equivalent activities to the four macrolides against strains of Streptococcus pyogenes. Streptococcus agalactiae and Moraxella catarrhalis. RP 59,500 exhibited the highest activities against Enterococcus faecalis, Enterococcus faecium and Enterococcus avium strains which are intrinsically resistant to most antimicrobial agents. No cross-resistance was observed between RP 59,500 and the four macrolides, which will merit attention in future clinical trials of the agent. The effect of human serum on the MIC of RP 59,500 was studied with strains of S. aureus, S. epidermidis and E. faecalis. The presence of 20% (V/V) serum had little or no effect on the MIC, although 50% (V/V) serum increased MICs by 4-8 folds. Laboratory-induced resistance to RP 59,500 occurred in a stepwise fashion in broth cultures of S. aureus, S. epidermidis and E. facalis strains and the induction rate was slow and no more than four fold increases were observed. Population analysis was performed on RP 59,500 and the reference macrolides against S. aureus ATCC 25,923 strain. Although low frequencies (less than 0.01%) of resistant sub-population were detected with EM, CAM, AZM and RXM, no RP 59,500-resistant sub-population was detected in this study. PMID- 9412876 TI - Coagulation tests predict bleeding after cardiopulmonary bypass. AB - OBJECTIVE: To determine the accuracy of coagulation profile laboratory tests, thromboelastography, and Sonoclot (SCT) values for predicting microvascular bleeding after cardiopulmonary bypass (CPB). DESIGN: A prospective, blinded trial. SETTING: A large academic medical center. PARTICIPANTS: Eighty-two adult patients undergoing elective cardiac surgery. INTERVENTIONS: Ten minutes after CPB, thromboelastography, SCT, and coagulation profile tests (bleeding time, prothrombin time, activated partial thromboplastin time, fibrinogen, fibrin split products, platelet count, mean platelet volume, and platelet hematocrit) were determined from a whole blood sample taken from an existing arterial catheter. Patients were subjectively defined as "bleeders" or "non-bleeders" by blinded clinical observers. Preoperative baseline tests were also obtained. MEASUREMENTS AND MAIN RESULTS: Thirty of the 82 patients (36.6%) were characterized as bleeders. Coagulation profile tests had the best correlation with intraoperative and postoperative blood loss. The specificity, sensitivity, and negative and positive predictive values were determined by receiver operating characteristic analysis, and the test values that differentiated normal from abnormal (bleeding) patients were determined. The coagulation profile laboratory tests had the greatest maximal sensitivity and specificity for predicting bleeding. These predictive values were outside the normal range for these laboratory tests. The thromboelastography values that produced maximal sensitivity and specificity were in the normal range for that test. CONCLUSION: Contrary to previous studies, coagulation profile tests had the greatest sensitivity and specificity to differentiate patients with excessive bleeding (abnormal) from those without excessive bleeding (normal) after CPB. Therefore, these tests should be used to guide transfusion therapy in patients who have excessive bleeding after CPB. PMID- 9412877 TI - Predictors of blood loss after coronary artery bypass grafting. AB - OBJECTIVE: To assess the predictive value of variables possibly associated with blood loss after coronary artery bypass grafting (CABG). DESIGN: A prospective study. SETTING: A university hospital. PARTICIPANTS: Eighty-nine patients scheduled for elective CABG. INTERVENTIONS: Blood samples were drawn before and after surgery. Chest tube drainage was measured hourly until removal of drains. MEASUREMENTS AND MAIN RESULTS: Activation of coagulation and fibrinolysis, routine clotting tests, and expression of platelet surface antigens were analyzed using flow cytometry. A significant correlation was found among blood loss and activated partial thromboplastin time, fibrinogen, prothrombin fragment 1 + 2, D dimers, platelet count, GPIb and P-selectin expression on platelets, use of internal thoracic artery, cross-clamp time, and thrombin-antithrombin III complex. In a multiple regression model, glycoprotein (GP) Ib expression on platelets, platelet count, use of internal thoracic artery, and D-dimers were significantly associated with blood loss. Logistic regression analysis showed that GPIb and D-dimers predicted an increased blood loss with a positive predictive value of 73% and a negative predictive value of 91%. CONCLUSIONS: Postoperative D-dimers and GPIb expression may be useful to exclude nonsurgical causes in bleeding patients after CABG. PMID- 9412878 TI - Platelet concentrate effects on thromboelastography. AB - OBJECTIVE: This study evaluated platelet effects on thromboelastography to determine how morphologically abnormal platelets affected native whole blood analysis. DESIGN: Prospective, controlled comparison. SETTING: Tertiary care university hospital. PARTICIPANTS: Volunteer cardiac surgery patients. INTERVENTIONS: Fresh platelets were obtained from volunteers and were either treated normally or cryodisrupted with liquid nitrogen. Fresh platelets, liquid nitrogen-treated platelets, or an equivalent quantity of the patient's blood were added to whole blood samples obtained from cardiac surgery patients before heparinization. Thromboelastographic parameters sensitive to platelet effects were measured in each of the three groups. MEASUREMENTS AND MAIN RESULTS: Maximum amplitude and alpha-angle significantly increased in the two groups receiving added platelets. There were no differences between the fresh platelet and the liquid nitrogen-treated platelet groups (Student's paired t-test). The R-time decreased significantly in both platelet-treated groups compared with the group that did not receive additional platelets. CONCLUSIONS: Viscoelastic changes in whole blood coagulation after the addition of platelet concentrates are not dependent on morphologically intact or functionally normal platelets. This in vitro study predicts that transfusion of poorly preserved platelet concentrates as well as fresh platelets would increase clot strength on thromboelastography if the recipient's blood were tested immediately after administration. PMID- 9412879 TI - Effect of aspirin in coronary artery bypass grafting. AB - OBJECTIVES: To evaluate the effect of aspirin (ASA) therapy on postoperative blood loss, transfusion requirements, reoperation for bleeding, duration of stay in the intensive care unit and in the hospital in a selected population undergoing a first coronary artery bypass grafting (CABG) surgery. DESIGN: Prospective observational study in consecutive patients during a 3-month period. SETTING: A teaching cardiothoracic center. PARTICIPANTS: Two hundred forty consecutive patients undergoing elective coronary artery bypass grafting surgery for the first time. INTERVENTIONS: Two hundred forty consecutive patients admitted for a first CABG the day before surgery were visited. patients with an abnormal routine coagulation screen or taking drugs that might have affected their coagulation mechanisms were prospectively excluded (n = 96). The date of the last dose of ASA was recorded in the 144 remaining patients, and data were acquired prospectively. MEASUREMENTS AND MAIN RESULTS: Total mediastinal blood drainage, blood products usage, reopening, and duration of intensive care unit and hospital stay were recorded. Patients were grouped by days free of ASA. There were no significant differences detected between groups. CONCLUSIONS: In patients undergoing a first CABG and with no known factors affecting their coagulation, ASA therapy did not appear to increase blood loss, reopening for bleeding, or blood products usage requirements during the hospital stay. ASA therapy did not influence the duration of stay in intensive care or in the hospital. PMID- 9412880 TI - A pump-prime aprotinin dose in cardiac surgery: appraisal of its effects on the hemostatic system. AB - OBJECTIVES: To examine pump-prime aprotinin action on coagulation and fibrinolysis in patients undergoing primary coronary revascularization. DESIGN: A prospective randomized study. SETTING: A university hospital. PARTICIPANTS: Forty three patients were randomly assigned to either group A, 21 patients treated with 2 x 10(6) kallikrein inhibitor units (KIU) of aprotinin in the cardiopulmonary bypass (CPB) prime, or group B, 22 patients, untreated. INTERVENTIONS: Patients, scheduled for elective coronary surgery, were treated with 2 x 10(6) KIU of aprotinin in the CPB prime. Markers of coagulation and fibrinolysis were evaluated. MEASUREMENTS AND MAIN RESULTS: Surgical times, number of reopenings, and allogeneic blood requirements were collected for each patient. Blood samples were obtained before and after surgery for assessing coagulation (prothrombin time [PT], activated partial thromboplastin time [aPTT], ethanol test, factor VII, antithrombin III [AT III], thrombin-antithrombin III complex [TAT], fragment 1.2 of prothrombin [F1.2]) and fibrinolysis (fibrin degradation products [FOP], plasmin-antiplasmin complexes [PAP], D-dimers) markers variations. In group A surgical times were faster, there were fewer reopenings (0 v 3), and fewer blood transfusions (1 patient v 4 patients). The two groups did not differ for PT, aPTT, and fibrinogen measurements. Postoperative FDP (measurable in more patients of group B at the end of the operation), PAP, and D-dimers postoperatory levels (less increased in aprotinin group) show the antifibrinolytic properties of the drug. Regarding the coagulation markers, factor VII decreased, whereas TAT and F1.2 increased, all to a lesser extent in the aprotinin group compared with the untreated patients, at the end of operation. CONCLUSION: Pump-prime aprotinin minimized, even if not completely inhibited, the activation of coagulation and fibrinolysis during CPB, possibly ensuring a less complicated and safer postoperative recovery. It seemed to allow the maintenance of a correct balance of hemostatic systems, avoiding the risk of thrombotic phenomena. PMID- 9412881 TI - Comparison of the effects of red cell separation and ultrafiltration on heparin concentration during pediatric cardiac surgery. AB - OBJECTIVE: To determine the effects of red cell separation and ultrafiltration on heparin concentration. DESIGN: Prospective study. SETTING: University-affiliated, pediatric medical center. PARTICIPANTS: Thirty-one children undergoing cardiac surgery. INTERVENTIONS: Blood sampled for heparin concentration and coagulation tests. MEASUREMENTS AND MAIN RESULTS: Thirteen infants under-went modified veno venous ultrafiltration (UF) after cardiopulmonary bypass (CPB). In addition, residual blood in the CPB circuit was hemoconcentrated by UF and reinfused (UF group). Heparin concentration increased from 2.0 +/- 0.6 to 2.5 +/- 0.8 U/mL, following modified UF; while activated coagulation time (ACT) decreased from 701 +/- 177 to 627 +/- 107 seconds. Heparin concentration of CPB circuit residual increased from 1.9 +/- 0.7 to 3.1 +/- 1.0 U/mL. In 18 children (older than 1 year old), the residual blood in the CPB circuit was hemoconcentrated by cell separation (CS) and reinfused (CS group). Heparin concentration of CPB circuit residual decreased from 2.6 +/- 0.6 to 0.3 +/- 0.2 U/mL. After reinfusion, patient heparin concentration remained unchanged at < 0.05 U/mL. Thrombin time increased from 28 +/- 6 to 48 +/- 29 seconds and did not correlate with H. CONCLUSIONS: The plasma concentration of heparin increased after veno-venous modified UF of the patient. Heparin concentration also increased after UF of residual CPB circuit blood. In contrast, circuit blood hemoconcentrated by CS contained minimal heparin, and, when infused, did not increase patient's heparin concentration. ACT and thrombin time did not correlate with heparin concentration. PMID- 9412882 TI - In vitro interference of the red cell substitute pyridoxalated hemoglobin polyoxyethylene with blood compatibility, coagulation, and clinical chemistry testing. AB - OBJECTIVES: Pyridoxalated hemoglobin-polyoxyethylene (PHP) is a prototypical red cell substitute approved for phase I studies. Peripheral blood smears of human blood mixed with PHP in 1 to 4 g/dL concentrations showed dose-dependent red cell aggregation and rouleaux. Whether this aggregation limits interpretation of blood compatibility testing and whether the intense coloration of serum or plasma containing PHP affects routine coagulation and clinical chemistry measurements was tested. DESIGN: In vitro studies. SETTING: University hospital laboratory. PARTICIPANTS: Four healthy volunteers, blood types A, B, AB, and O. All were Rh+. MEASUREMENTS AND MAIN RESULTS: ABO typing, Rh typing, and antibody screening and coagulation studies were performed on blood: PHP admixtures having final concentrations of 1, 2, and 4 g/dL. For clinical chemistry interference studies, known concentrations of analytes were added to a serum matrix containing PHP. ABO (forward) and Rh typing showed no interference in the three concentrations tested. Reverse ABO typing and antibody screening showed rouleaux at 4 g/dL, which corrected with routine saline replacement. Partial thromboplastin time (PTT), prothrombin time (PT), and fibrinogen showed no clinically significant differences from the controls. Results for electrolytes, renal function analytes, and markers of cardiac injury were acceptable by standard laboratory methods. However, results of liver function tests were unacceptable in PHP-containing specimens. CONCLUSIONS: PHP-induced aggregation was observed with high PHP concentration; however, compatibility testing was not affected because agglutination was corrected by saline replacement, which is standard practice. Although routine blood banking, coagulation, and most clinical chemistry analytes can be measured reliably, alternative methods and strategies are needed for assessing liver function in the presence of PHP. PMID- 9412883 TI - The incidence of artery puncture with central venous cannulation using a modified technique for detection and prevention of arterial cannulation. AB - BACKGROUND: Cannulation of the central circulation is essential for management of patients who require major surgery, and for patients who are critically ill. Arterial puncture is the most frequent complication associated with central venous cannulation, and is potentially fatal. Detection of arterial puncture can be problematic, especially in patients with cyanotic congenital heart disease. METHODS: One thousand eleven consecutive cardiothoracic and vascular surgical patients who required central venous cannulation were studied using a new technique for detection of arterial puncture and prevention of arterial cannulation. This technique involves continuous pressure transduction of the steel introducer needle. Central venous cannulation was attempted in all patients. The sites of attempted catheterizations, number of arterial punctures and cannulations, and the number of successful catheterizations were noted. All patients were treated in accordance with standard anesthetic and surgical techniques in the institution. RESULTS: One thousand one hundred seventy-two central venous catheters were placed. The overall success rate was 99.6%. The incidence of arterial puncture was 9.3% for central venous cannulation attempts of the internal jugular, subclavian, and femoral veins. No arterial cannulation occurred, and none of the patients had significant complications. Congenital heart disease patients had a higher incidence of arterial puncture (14.1%) and a lower rate (96.8%) of successful cannulation. CONCLUSION: Pressure transduction of the steel needle is a useful technique for detecting arterial puncture and preventing arterial cannulation during attempts to achieve central venous cannulation. PMID- 9412884 TI - Association between R-wave amplitude of the electrocardiogram and myocardial function during coronary artery bypass grafting. AB - OBJECTIVE: The recovery of R-wave amplitude in the V5 lead of the electrocardiogram (ECG) was recently found to be worse in nonsurvivors than in survivors after coronary artery bypass grafting (CABG). On the contrary, an increase in R-wave amplitude has been found to reflect myocardial dysfunction in exercise testing. The purpose of this study was to examine whether the changes in R-wave amplitude are associated with changes of myocardial function during CABG. DESIGN: A prospective clinical study. SETTING: Cardiothoracic division of surgery in a university hospital. PARTICIPANTS: Ten consecutive patients undergoing CABG. MEASUREMENTS: R-wave amplitude was measured at eight different time points. Left ventricular end-systolic wall tension, wall stress at isovolumic contraction (afterload), end-diastolic wall stress (preload), end-systolic wall stress per end-systolic area (contractility), and stroke work were calculated using transesophageal echocardiography and arterial pressure. MAIN RESULTS: Linear regression was calculated between changes in R-wave amplitude and echo parameters. A weak positive association within subjects was noted among R amplitude and all measured cardiac function parameters except preload. R2 value varied from 0.101 to 0.266, and R2 for preload was 0.017. CONCLUSIONS: These results suggest that only 10% to 27% of variation in R-wave amplitude can be explained by left ventricular function indices measured by echocardiography in patients with CABG. Thus, R-wave amplitude changes in an individual patient undergoing CABg have very limited utility as a noninvasive measure of left ventricular function. PMID- 9412885 TI - Nitroglycerin-induced hypoxemia does not produce myocardial ischemia. AB - OBJECTIVE: Nitroglycerin has been the drug of choice for relieving myocardial ischemia for more than a hundred years. Several studies have indicated that a significant reduction in arterial oxygen tension (PaO2) occurs after the administration of sublingual nitroglycerin to patients with coronary artery disease breathing room air. Because available oxygen in arterial blood is reduced, it would be reasonable to assume that oxygen delivery to the myocardium would also be impaired. The purpose of this study was to investigate whether nitroglycerin-induced arterial desaturation results in compromised oxidative metabolism of myocardium assessed by coronary sinus lactate concentration and oxygen content in patients with coronary artery disease undergoing coronary artery bypass surgery. PARTICIPANTS: Ten randomly selected patients undergoing coronary bypass surgery. SETTING: All studies were performed at Siyami Ersek Cardiovascular and Thoracic Surgery Center. METHODS: A catheter was inserted into the radial artery to measure blood gases and arterial lactate concentration. After sternotomy, and aortic and venous cannula placement, a coronary sinus catheter was introduced into the coronary sinus to measure oxygen content and lactate concentration. Control coronary sinus and arterial blood samples were obtained before nitroglycerin infusion. Nitroglycerin was then given in a dose of 2 micrograms/kg/min for a period of 5 minutes. At the end of 5 minutes, second samples were obtained from the coronary sinus and arterial catheters. MAIN RESULTS: It was found that arterial and coronary sinus oxygen tension decreased significantly. Arterial lactate concentration did not change, coronary sinus lactate concentration decreased. Despite a substantial fall in arterial oxygen tension after administration of nitroglycerin, a significant reduction in coronary sinus lactate concentration occurred. CONCLUSION: Nitroglycerin-induced hypoxia does not compromise oxidative metabolism of myocardium as can be assessed by a concomitant decrease in coronary sinus lactate concentration. PMID- 9412886 TI - Effects of calcium on left ventricular function early after cardiopulmonary bypass. AB - OBJECTIVES: Evaluation of the effects of intravenous CaCl2 on systolic and diastolic function early after separation from cardiopulmonary bypass (CPB) DESIGN: Prospective study SETTING: University hospital PARTICIPANTS: Twenty patients scheduled for elective coronary artery surgery INTERVENTIONS: Left ventricular (LV) pressures were measured with fluid-filled catheters. Data were digitally recorded during pressure elevation induced by tilt-up of the legs. Transgastric short-axis echocardiographic views of the LV were simultaneously recorded on videotape. Measurements were obtained before the start of CPB, 10 minutes after termination of CPB, after intravenous administration of CaCl2, 5 mg/kg, and 10 minutes later. MEASUREMENTS AND MAIN RESULTS: Systolic function was evaluated with the slope (Ees, mmHg/mL) of the systolic pressure-volume relation. Diastolic function was evaluated with the chamber stiffness constant (Kc, mmHg/mL) of the diastolic pressure-volume relation. CaCl2 increased Ees from 2.62 +/- 0.46 to 5.58 +/- 0.61 (mean +/- SD), but induced diastolic dysfunction with an increase in Kc from 0.011 +/- 0.006 to 0.019 +/- 0.007. These changes were transient and had disappeared within 10 minutes after administration of CaCl2. CONCLUSIONS: CaCl2 early after CPB transiently improved systolic function at the expense of an increase in ventricular stiffness, suggesting temporary diastolic dysfunction. PMID- 9412887 TI - Amrinone versus dopamine-nitroglycerin after reconstructive surgery for complete atrioventricular septal defect. AB - OBJECTIVES: To compare the effectiveness and safety of amrinone and a combination of dopamine and nitroglycerin in infants after reconstructive surgery for congenital heart disease. DESIGN: A prospective, randomized, double-blind study. SETTING: Pediatric intensive care unit in a university hospital. PARTICIPANTS: Thirty-two infants with complete atrioventricular septal defect. INTERVENTIONS: Amrinone loading dose, 2 mg/kg, followed by a maintenance infusion, 7.5 micrograms/kg/min, was given to 17 infants before separation from cardiopulmonary bypass. The remaining 15 patients received a combination of dopamine, 5 micrograms/kg/min, and nitroglycerin, 1 microgram/kg/min. MEASUREMENTS AND MAIN RESULTS: The circulatory state of the patients was evaluated from 4 to 18 hours after cardiopulmonary bypass. The systemic blood flow index, calculated using the Fick principle, was higher in the amrinone group (2.5 +/- 0.7 L/min/m2) compared with the dopamine-nitroglycerin group (2.0 +/- 0.6 L/min/m2, mean +/- SD). The pulmonary blood flow index in the amrinone group was higher (2.9 +/- 0.6 L/min/m2) than in the dopamine-nitroglycerin group (2.2 +/- 0.6 L/min/m2); no significant difference was noted in the mean pulmonary artery pressure. The oxygen extraction ratio was higher in the dopamine-nitroglycerin group (0.41 +/- 0.07) compared with the amrinone group (0.34 +/- 0.08). Despite lower platelet counts in the amrinone group, no hemorrhagic complications were seen in any patient. CONCLUSIONS: With this dosage regimen, amrinone provides a higher cardiac output, more favorable oxygen dynamics, and lower pulmonary vascular resistance than dopamine and nitroglycerin. PMID- 9412888 TI - Coagulation complicating cardiopulmonary bypass in a patient with heparin-induced thrombocytopenia receiving the heparinoid, danaparoid sodium. PMID- 9412889 TI - Management of anticoagulation during cardiopulmonary bypass in a patient with a circulating lupus anticoagulant. PMID- 9412890 TI - Posttransplant primary heart dysfunction and myocardial stunning. PMID- 9412892 TI - Validity of pulmonary artery catheter-derived hemodynamic information during bronchopulmonary lavage. PMID- 9412891 TI - The use of neostigmine to decrease the heart rate in a patient undergoing minimally invasive coronary artery bypass surgery. PMID- 9412893 TI - Venous thrombosis in pediatric cardiac surgery. PMID- 9412894 TI - Progress in cardiac arrhythmia ablation: potential for broader application and shorter procedure time. AB - Catheter ablation is a very effective form of therapy for cardiac arrhythmias. Using present techniques, however, its application is limited only to a select type of arrhythmia involving discrete areas of the heart. The more common arrhythmias, such as atrial fibrillation and ventricular tachycardia, remain difficult to treat. It seems likely that deeper and larger ablations would be necessary for abolishing reentry substrates of VT and that long linear ablations would be needed for preventing propagation of atrial fibrillation. Improvements in energy delivery techniques and the use of alternative energy modalities are promising developments that may yield a broader scope of lesion generation. Cooling mechanisms have allowed for more effective RF delivery and the generation of larger lesions. Laser, ultrasound, and microwave ablations have also been shown to produce deeper lesions. Linear lesions could be accomplished with RF ablation using multiple electrode configurations or with MW using long antennae. Future ablation procedures are likely to include complex arrhythmias and use one of these newer modalities. The more powerful and versatile modalities may become the most useful ones as they, coupled with parallel improvements in mapping technology, would facilitate the procedures by minimizing morbidity and improving patients' tolerance. At present, the more simple procedures do not require anesthesia or intensive cardiorespiratory monitoring, but because the procedures are being applied to include more complex arrhythmias, such monitoring may become necessary. PMID- 9412895 TI - Case 5-1997. Successful resuscitation of a patient with severe accidental hypothermia and prolonged cardiocirculatory arrest using cardiopulmonary bypass. PMID- 9412896 TI - Pro: calcium is routinely indicated during separation from cardiopulmonary bypass. PMID- 9412897 TI - Con: calcium is not routinely indicated during separation from cardiopulmonary bypass. AB - Severe hypocalcemia is uncommon in adult cardiac surgery patients; the nearly ubiquitous mild hypocalcemia does not impair myocardial performance. Clinicians should recognize that in certain circumstances, calcium may interact negatively with catecholamines such as epinephrine or dobutamine. Lastly, evidence suggests that calcium influx during ischemia-reperfusion contributes to myocardial dysfunction after CPB. Therefore, there appears to be no justification for the practice of routinely administering large doses of calcium salts to adult cardiac surgery patients after CPB. PMID- 9412898 TI - An unusual echocardiograph after aortic valve replacement. Aortic dissection. PMID- 9412899 TI - Hemodynamics of aortic cross-clamping. PMID- 9412900 TI - Calcium channel blockers for ischemia after cardiopulmonary bypass. PMID- 9412902 TI - Selecting the correct size left double-lumen tube. PMID- 9412901 TI - Method of choice for measuring mitral regurgitation. PMID- 9412903 TI - Thoracic surgery and difficult intubation: another application of univent tube for one-lung ventilation. PMID- 9412904 TI - Inhaled nitric oxide and one-lung ventilation in the lateral decubitus position. PMID- 9412905 TI - Differential plasma membrane distribution of metabotropic glutamate receptors mGluR1 alpha, mGluR2 and mGluR5, relative to neurotransmitter release sites. AB - Two group I metabotropic glutamate receptor subtypes, mGluR1 and mGluR5, have been reported to occur in highest concentration in an annulus surrounding the edge of the postsynaptic membrane specialisation. In order to determine whether such a distribution is uniform amongst postsynaptic mGluRs, their distribution was compared quantitatively by a pre-embedding silver-intensified immunogold technique at electron microscopic level in hippocampal pyramidal cells (mGluR5), cerebellar Purkinje cells (mGluR1 alpha) and Golgi cells (mGluR2). The results show that mGluR1 alpha, mGluR5 and mGluR2 each have a distinct distribution in relation to the glutamatergic synaptic junctions. On dendritic spines, mGluR1 alpha and mGluR5 showed the highest receptor density in a perisynaptic annulus (defined as within 60 nm of the edge of the synapse) followed by a decreasing extrasynaptic (60-900 nm) receptor level, but the gradient of decrease and the proportion of the perisynaptic pool (mGluR1 alpha, approximately 50%; vs mGluR5, approximately 25%) were different for the two receptors. The distributions of mGluR1 alpha and mGluR5 also differed significantly from simulated random distributions. In contrast, mGluR2 was not closely associated with glutamatergic synapses in the dendritic plasma membrane of cerebellar Golgi cells and its distribution relative to synapses is not different from simulated random distribution in the membrane. The somatic membrane, the axon and the synaptic boutons of the GABAergic Golgi cells also contained immunoreactive mGluR2 that is not associated with synaptic specialisations. In the hippocampal CA1 area the distribution of immunoparticles for mGluR5 on individual spines was established using serial sections. The results indicate that dendritic spines of pyramidal cells are heterogeneous with respect to the ratio of perisynaptic to extrasynaptic mGluR5 pools and about half of the immunopositive spines lack the perisynaptic pool. The quantitative comparison of receptor distributions demonstrates that mGluR1 alpha and mGluR5, but not mGluR2, are highly compartmentalised in different plasma membrane domains. The unique distribution of each mGluR subtype may reflect requirements for different transduction and effector mechanisms between cell types and different domains of the same cell, and suggests that the precise placement of receptors is a crucial factor contributing to neuronal communication. PMID- 9412906 TI - Double bouquet cell axons in the human temporal neocortex: relationship to bundles of myelinated axons and colocalization of calretinin and calbindin D-28k immunoreactivities. AB - We have examined the distribution of double bouquet cell axons, immunocytochemically stained for the calcium-binding proteins calretinin and calbindin D-28k in the human temporal neocortex, in relation to bundles of myelinated axons (originating from pyramidal cells) and the colocalization of these calcium-binding proteins. The large number and regularity of distribution of double bouquet cell axons was clearly visualized in tangential sections from cortical layers III--V. In these sections, we estimated that the mean number +/- standard deviation of double bouquet cell axons per 10,000 microns2 was 11.65 +/- 0.44 with a mean diameter of 12.10 +/- 0.63 microns and a mean center-to-center spacing of 29.8 +/- 0.91 microns. These values are very similar to those previously reported in the monkey neocortex. The distribution of double bouquet cell axons was closely related to bundles of myelinated axons; there was overlapping with basically a one-to-one correspondence. Finally, double-label immunofluorescence experiments revealed that the vast majority of double bouquet cell axons immunoreactive for calbindin were also stained for calretinin. Since relatively few cell somata were double-labeled in the human temporal cortex, we concluded that double bouquet cells may represent a significant subpopulation of neurons that colocalize these calcium-binding proteins. PMID- 9412907 TI - Calretinin- and parvalbumin-immunoreactive neurons in the rat main olfactory bulb do not express NADPH-diaphorase activity. AB - The presence of nitric oxide synthase (NOS) in neuronal elements expressing the calcium-binding proteins calretinin (CR) and parvalbumin (PV) was studied in the rat main olfactory bulb. CR and PV were detected by using immunocytochemistry and the nitric oxide (NO) -synthesizing cells were identified by means of the reduced nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-diaphorase) direct histochemical method. The possible coexistence of NADPH-diaphorase and each calcium-binding protein marker was determined by sequential histochemical immunohistochemical double-labeling of the same sections. Specific neuronal populations were positive for these three markers. A subpopulation of olfactory fibers and olfactory glomeruli were positive for either NADPH-diaphorase or CR. In the most superficial layers, groups of juxtaglomerular cells, superficial short-axon cells and Van Gehuchten cells demonstrated staining for all three markers. In the deep regions, abundant granule cells were NADPH-diaphorase- and CR-positive and a few were PV-immunoreactive. Scarce deep short-axon cells demonstrated either CR-, PV-, or NADPH-diaphorase staining. Among all these labeled elements, no neuron expressing CR or PV colocalized NADPH-diaphorase staining. The present data contribute to a more detailed classification of the chemically- and morphologically-defined neuronal types in the rodent olfactory bulb. The neurochemical differences support the existence of physiologically distinct groups within morphologically homogeneous populations. Each of these groups would be involved in different modulatory mechanisms of the olfactory information. In addition, the absence of CR and PV in neuronal groups displaying NADPH-diaphorase, which moreover are calmodulin-negative, indicate that the regulation of NOS activity in calmodulin-negative neurons of the rat olfactory bulb is not mediated by CR or PV. PMID- 9412909 TI - Interferon therapy for chronic HCV hepatitis: trick or treat? AB - Chronic hepatitis caused by the hepatitis C virus (HCV) is a common condition that leads to cirrhosis and hepatocellular carcinoma. Current treatment with interferon is unsatisfactory, with a low percentage of patients who respond and uncertain high-term significance; in addition, it is associated with sometimes severe side effects. The increasing sophistication of molecular biology has enabled viral characteristics such as viral load, genotypes, and quasi-species to be identified, which may help predict a patient's response to interferon treatment. We suggest that interferon therapy for hepatitis C virus should be restricted to referral centers in the context of controlled trials. PMID- 9412908 TI - Association between pituitary adenylate cyclase-activating polypeptide and thyrotropin-releasing hormone in the rat hypothalamus. AB - Pituitary adenylate cyclase-activating polypeptide (PACAP) is present in many regions of the hypothalamus including the paraventricular nucleus (PVN). In this study the anatomical relationship between PACAP- and thyrotropin-releasing hormone (TRH)-immunoreactive neuronal elements was investigated in the rat hypothalamus. Using a well-characterized mouse monoclonal antibody against PACAP and a rabbit polyclonal antiserum against TRH, we found numerous nerve fibers with PACAP-immunoreactivity (ir) closely apposed to TRH neurons in the PVN suggesting synaptic contacts. Electron microscopy confirmed the presence of synapses between PACAP-ir terminals and TRH-ir perikarya and various dendritic profiles as well as between PACAP-ir terminals and unlabeled perikarya and small- to medium-sized dendrites. Coexistence of the two peptides in perikarya of the PVN was limited to only a few neurons in the periventricular subdivision, but PACAP-ir and TRH-ir extensively coexisted in perikarya of the perifornical cell group, medial preoptic area, lateral hypothalamus and dorsomedial nucleus. The interactions between PACAP-containing neuronal processes and TRH neurons in the PVN raise the possibility that PACAP modulates the secretion of TRH destined for regulation of anterior pituitary TSH. The more general association between PACAP and TRH in other regions of the hypothalamus suggests a further role for PACAP as a cofactor in the function of TRH neurons. PMID- 9412910 TI - Exclusion of the meal period improves the clinical reliability of esophageal pH monitoring. AB - Meal period exclusion from 24-h pH testing allows better separation between controls and patients with gastroesophageal reflux disease. We reviewed the results of 24-h pH studies of 350 patients with reflux symptoms. They were divided into two groups based on the 95th percentile of the total percentage of time when pH was < 4 for healthy persons in our laboratory. Thus group A consisted of 212 patients with symptoms and normal acid exposure and group B consisted of 138 patients with symptoms and abnormal acid exposure. The change in upright reflux excluding the meal period was calculated for each patient. Meal period exclusion resulted in opposite effects for the two groups of patients, with a change in median upright reflux of -0.6% for group A and +0.5% for group B (p < 0.0001). After meal exclusion, five patients were reclassified as having reflux, with four (80%) of these responding to antireflux therapy. Nine other patients were recategorized as not having reflux after meal exclusion. Only one of seven patients (14%) for whom data were available responded to treatment (two patients were lost to follow-up). We recommend meal period exclusion from pH analysis because it improves the clinical reliability of esophageal pH monitoring. PMID- 9412911 TI - Changes in serum pepsinogen, gastrin, and immunoglobulin G antibody titers in helicobacter pylori-positive gastric ulcer after eradication of infection. AB - There are no studies of changes in immunoglobulin G (IgG) titers to Helicobacter pylori, serum pepsinogen, and gastrin in patients with H. pylori-positive gastric ulcers. We investigated the effect of therapy for H. pylori-positive gastric ulcer on IgG titers to H. pylori, serum pepsinogen I and II, and gastrin. Thirty six patients with H. pylori-positive gastric ulcer were treated with lansorazole and antibiotics for 2 weeks. Serum pepsinogen I and II concentrations, serum gastrin, and IgG titers to H. pylori were measured before treatment and then at 4 and 12 weeks after stopping the treatment. The presence or eradication of H. pylori was determined using the rapid urease test and by histologic H. pylori staining. For 19 patients in whom H. pylori had been successfully eradicated, the pepsinogen I/II ratio increased, pepsinogen II levels decreased, and the anti-H. pylori IgG decreased compared with the results from before therapy and with those from 4 and 12 weeks after therapy. Gastrin levels decreased compared with pretreatment results and those from 4 weeks after the end of treatment. In 17 patients in whom the therapy failed to eradicate H. pylori infection, there were no sequential significant changes in the pepsinogen I/II ratio or in the levels of pepsinogen I, pepsinogen II, anti-H. pylori IgG, and gastrin. A decrease in the serum levels of the IgG antibody to H. pylori and gastrin and also an increase in the pepsinogen I/II ratio could be used as predictors for the eradication of H. pylori infection in gastric ulcer. PMID- 9412912 TI - Effect of ranitidine on the urea breath test: a controlled trial. AB - Because Helicobacter pylori is an acid-sensitive organism, an elevation of the gastric pH by H2 inhibitors might improve the intragastric conditions for the development of this organism. We tested this hypothesis in a prospective and controlled trial including 43 patients positive for H. pylori using the rapid urease test. Twenty-six patients received 150 mg ranitidine twice daily and 17 patients received no treatment. The 14C-urea breath test was performed in both groups at the beginning of the study and 2 weeks later. Radioactive 14C in exhaled carbon dioxide was significantly increased (p = 0.045) in the patients treated with ranitidine, compared with the patients in the control group. Administration of this drug to patients infected with H. pylori is associated with an increase in the bacterial load after 2 weeks of treatment. This phenomenon might be attributed to increased bacterial growth due to the H2 blocker. PMID- 9412913 TI - High prevalence of celiac sprue among patients with primary biliary cirrhosis. AB - Although coexisting primary biliary cirrhosis (PBC) and celiac sprue have been described, celiac sprue is sufficiently common in western Europe for chance to explain isolated cases. We screened our patients with PBC for celiac sprue using serum immunoglobulin A endomysial antibody (EmA), with confirmation by duodenal biopsy in EmA-positive patients. Of 57 patients, 6 (11%) had EmA. Four agreed to have a biopsy taken, and all had villous atrophy, yielding a minimum prevalence of 1:14 (7%). Apart from anemia in one patient, none of the four had symptoms or routine laboratory abnormalities suggestive of celiac sprue. None had improvement in liver biochemical tests after 12 to 24 months on gluten-free diets despite the disappearance of EmA. Celiac sprue is common among patients with PBC and they should be routinely screened for this condition. Symptoms wrongly attributed to PBC may respond to gluten exclusion, and both conditions are potent risk factors for osteoporosis. PMID- 9412914 TI - Azathioprine in steroid-resistant and steroid-dependent ulcerative colitis. AB - We evaluated, retrospectively, the outcome of 56 patients (39 male, 17 female; mean age, 34 years; age range, 14-65 years) who received azathioprine for either steroid-resistant (group A, n = 10) or steroid-dependent (group B, n = 46) ulcerative colitis. The patients were followed for a mean of 29 +/- 17 months (median, 27 months). Twenty-four had left-sided colitis, 5 had subtotal colitis, and 27 had total colitis. The mean duration of the disease was 51 months (range, 2-192 months). At the beginning of azathioprine treatment (time 0), all patients had clinically severe disease and were taking 40 mg prednisolone per day. Azathioprine was used in addition to steroid therapy at a dosage of 2 mg/kg. The need for steroids, expressed as the median cumulative steroid dose (mg/year), and the number of clinical relapses (requiring steroid therapy) in the 2 years before azathioprine treatment, were compared with those of the 3-year follow-up with azathioprine treatment. A positive response to azathioprine was defined as (a) avoidance of colectomy, (b) a significant decrease in the median cumulative steroid dose, and (c) a significant decrease in the number of clinical relapses (expressed as number/patient/year). One patient in group A withdrew due to painful dyspepsia, and two patients in group B were lost to follow-up. Remission with complete elimination of steroids was achieved in 36 of 53 (64%), 23 of 35 (66%), and 18 of 26 (69%) patients in the first, second, and third years, respectively, of azathioprine treatment. Compared with the 2 years before azathioprine treatment, a significant decrease was observed of about 75% both in steroid consumption and in the number of clinical relapses during the 3 years of azathioprine therapy. Two of nine patients in group A and 2 of 44 patients in group B had colectomy after mean periods of 15 months and 24 months, respectively. Azathioprine is effective and safe in avoiding colectomy in patients with steroid-resistant and steroid-dependent ulcerative colitis; its use decreases both steroid requirements and clinical relapses. PMID- 9412915 TI - Desmoid tumors in familial adenomatous polyposis/Gardner's syndrome. AB - To clarify the clinical risk of desmoid tumors developing in familial adenomatous polyposis, we reviewed the cases of 49 Japanese patients diagnosed with familial adenomatous polyposis at our institute. In six patients who manifested desmoid tumors at a mean age of 31 years, we reviewed the clinical features and compared various phenotypic manifestations with those in the 43 patients without desmoid tumors. During the observation periods (mean, 6.5 years), two of six patients with desmoid tumors died because of the tumors, which measured > 10 cm in diameter at the initial diagnosis, whereas the remaining four patients with desmoid tumors < 5 cm did not experience complications. The patients with desmoid tumors tended to be women (5 of 6 vs. 17 of 43; p = 0.05) and more frequently had gastric fundic gland polyposis (5 of 6 vs. 17 of 43; p = 0.05) than did the patients without desmoid tumors. There were no apparent differences in other clinical manifestations, including the incidences of colonic polyposis, gastroduodenal adenomas, and extraintestinal tumors. Desmoid tumors can be serious complication in patients with familial adenomatous polyposis. There may be some association in the genesis of desmoid tumors and gastric fundic gland polyposis. PMID- 9412916 TI - Ileal endometriosis masquerading as Crohn's ileitis. AB - Endometriosis is usually confined to the pelvis but may involve distant organs. When the small bowel is affected, endometriosis has a propensity to develop in the distal ileum, which may lead to fibrosis and stricture formation that can be confused with Crohn's disease. Here were describe two women, one with antecedent Crohn's colitis in whom ileal endometriosis mimicked obstructing Crohn's disease of the terminal ileum. These reports illustrate that ileal endometriosis must be considered in the differential diagnosis of Crohn's disease of the ileum, even in the presence of Crohn's disease elsewhere in the gastrointestinal tract. PMID- 9412917 TI - Effect of relaxation music on patient tolerance of gastrointestinal endoscopic procedures. AB - The use of relaxation music as an adjunct to sedation has not been well studied. We tried to determine whether the use of relaxation music can improve patient tolerance of gastrointestinal endoscopic procedures. Fifty-nine patients undergoing gastrointestinal endoscopic procedures were randomly assigned to receive either relaxation music (n = 28) or no music (n = 31) using headphones and a portable compact disc player. Patient anxiety before the procedure, tolerance of the procedure, and willingness to undergo a repeated procedure were self-assessed using a visual analog scale. Patient tolerance was also assessed by the assisting nurse. There was no significant difference in the overall tolerance score between the two groups. However, a significantly higher proportion of patients described the experience of a gastrointestinal endoscopic procedure as being at least moderately unpleasant in the no-music group. Patient acceptance of the relaxation music was high: 82% in the group stated they would have music again if they required another procedure. We conclude that, even in patients who have sedation, relaxation music can reduce the number who find the experience of gastrointestinal endoscopic procedures unpleasant. Therefore we believe it has a role as an adjunct to sedation in gastrointestinal endoscopic procedures. PMID- 9412919 TI - Can pressure overload cause sliding hiatal hernia? A case report and review of the literature. AB - We describe a hiatal hernia of moderate size in a 31-year-old competitive bodybuilder to raise the question of whether such hernias are more likely in young elite resistance-trained athletes as a consequence of attempts to increase intra-abdominal pressure and thus decrease the strain on the lumbar spine. PMID- 9412918 TI - Virus elimination and histologic improvement in patients with chronic hepatitis C treated with interferon alpha. AB - We evaluated the elimination of hepatitis C virus (HCV) by means of interferon alpha (IFN-alpha) by investigating both positive- and negative-stranded RNA forms in the peripheral blood mononuclear cells and liver tissue. We also assessed the long-term histologic improvement accompanying viral clearance. We studied 20 persons with HCV whose serum aminotransferase levels remained normal for more than 1.5 years after IFN-alpha treatment withdrawal. The presence of HCV RNA in their peripheral blood and of both positive and negative strands in the peripheral blood mononuclear cells and liver tissue was investigated using the reverse transcription polymerase chain reaction method. We examined the histologic findings using the European classification and the histology activity index scoring system. In 16 of 20 patients, both strands disappeared from the possible reservoirs. The histologic findings indicated reduced activity, and histology activity index scores (1, 2, 3, and total) also showed significant improvement. We confirmed that IFN-alpha therapy can induce the elimination of HCV RNA from the conceivable HCV reservoirs and effect histologic improvement. Therapy with IFN-alpha is effective for treating chronic hepatitis C. PMID- 9412920 TI - Massive bleeding from multiple gastric ulcerations in a patient with lymphocytic gastritis and celiac sprue. AB - Uncontrolled hemorrhage and multisystem organ failure developed in a patient with celiac sprue, lymphocytic gastritis, and diffuse gastric ulceration. A proximal small bowel biopsy showed villous atrophy and lymphoplasmacytic infiltration consistent with celiac sprue. At autopsy, there were no gross or histologic findings to suggest lymphoma. The intestinal lymphocytic infiltrate was not monoclonal, and gene rearrangements were not detected. Lymphocytic gastritis is a rare cause of upper gastrointestinal hemorrhage, which may be the result of sensitivity to gluten or other luminal antigens. This diagnosis should be considered in cases of diffuse gastric ulceration with bleeding in which the endoscopic appearances are not typical of peptic ulcer disease or drug-induced erosions. Ideally, biopsies of gastric and duodenal mucosa should be performed to establish the diagnosis. PMID- 9412922 TI - A case of systemic malignant lymphoma with intestinal involvement of lymphomatous polyposis type. AB - Multiple lymphomatous polyposis is a rare type of intestinal lymphoma characterized by non-Hodgkin's lymphoma of follicular mantle cell origin and extremely poor prognosis. We report a case of systemic lymphoma with the intestinal involvement of multiple lymphomatous polyposis. Although radiographic and endoscopic features of the case were compatible with multiple lymphomatous polyposis, histologic evidence suggested the diagnosis of diffuse large cell lymphoma rather than mantle cell lymphoma. Our case seems to be unique in its histologic findings and also in its prognosis, because the patient has been alive for more than 50 months after diagnosis. PMID- 9412921 TI - Ileocecal intussusception of small-bowel lymphoma: diagnosis by colonoscopy. AB - Intussusception is rare in adults. There is little information on the role of colonoscopy in colonic intussusception. We report, to our knowledge, the first adult case of small-bowel lymphoma causing ileocecal intussusception in which the diagnosis was made by colonoscopy. Colonoscopy has a useful role in the diagnosis and management of ileocecal intussusception. PMID- 9412923 TI - Chronic granulocytic leukemia in a patient with ankylosing spondylitis and ulcerative colitis: an interesting association. AB - Chronic granulocytic leukemia in a 59-year-old man with ankylosing spondylitis and ulcerative colitis is described. Ankylosing spondylitis was confirmed at age 28 years and ulcerative colitis at age 49 years. Etiologic considerations and a brief review of the literature are presented. PMID- 9412924 TI - Acute colitis associated with etodolac. AB - We describe two patients in whom acute colitis developed during etodolac treatment. Symptoms resolved when etodolac was stopped. In one, symptoms of colitis recurred with reexposure to the drug. The clinical and pathologic findings were consistent with de novo colitis from etodolac. PMID- 9412925 TI - Hyperplastic (metaplastic) polyposis of the colorectum associated with adenomas and an adenocarcinoma. AB - Hyperplastic (metaplastic) polyposis associated with adenoma and adenocarcinoma of the colorectum is rare. We describe a 55-year-old man with hyperplastic polyposis associated with multiple adenomas and an adenocarcinoma who underwent total colectomy. We found at least 200 polyps in the surgical specimen. Nearly all of the polyps were hyperplastic, and some were adenomas. Furthermore, some hyperplastic polyps had adenomatous areas. This indicates the transformational sequence of a hyperplastic polyp to adenoma to adenocarcinoma. PMID- 9412926 TI - Lower gastrointestinal bleeding from a hepatocellular carcinoma invading the colon. AB - Bleeding from the gastrointestinal tract due to hepatocellular carcinoma invasion is unusual. We describe a 71-year-old man who had bloody stools caused by a hepatocellular carcinoma that directly invaded the transverse colon. The diagnosis was confirmed by colonoscopy and tissue examination. Our patient is the first with lower gastrointestinal bleeding from a hepatocellular carcinoma during the natural course of the tumor. PMID- 9412927 TI - Is melatonin associated with the development of autoimmune hepatitis? AB - Melatonin is a neurohormone produced by the human pineal gland that plays a role in the regulation of many physiologic processes and has been proposed as a therapy for everything from insomnia to metastatic carcinoma. Melatonin is available in the United States without prescription, and adverse effects appear to be uncommon. However, because melatonin appears to have immunomodulatory properties, the potential exists for the development of autoimmune-related side effects. We describe a patient in whom characteristic clinical and laboratory features of autoimmune hepatitis developed after beginning melatonin therapy for the treatment of insomnia. Liver biopsy demonstrated histologic features of autoimmune hepatitis. Rapid symptomatic and biochemical improvement resulted from the initiation of immunosuppressive therapy; however, hepatitis recurred after the withdrawal of steroid therapy. The temporal relation observed between melatonin use and the development of autoimmune hepatitis raises the possibility that the drug might be involved in the pathogenesis of this patient's autoimmune disease. PMID- 9412928 TI - Diffuse fatty liver in familial heterozygous hypobetalipoproteinemia. AB - A 34-year-old man had asymptomatic hepatomegaly, slightly increased serum alanine aminotransferase and gamma-glutamyl transpeptidase levels, and a sonographic pattern suggesting diffuse hepatic steatosis. Liver biopsy revealed fatty change in 25% to 50% of hepatocytes. The patient also had low serum levels of cholesterol and triglycerides and met clinical, biochemical, and familial diagnostic criteria of heterozygous hypobetalipoproteinemia. We could not relate his hepatic steatosis to any already known cause of fatty liver and could only attribute it to heterozygous hypobetalipoproteinemia. Familial heterozygous hypobetalipoproteinemia should be ruled out in patients with unexplained hepatic steatosis. PMID- 9412929 TI - Disseminated intra-abdominal cystic lymphangiomatosis with severe intestinal bleeding. A case report. AB - We describe cystic lymphangiomatosis with intestinal bleeding developing multiple lymphangiomas in the small intestine, mesentery, mesocolon, omentum, retroperitoneum, and spleen. Small intestinal fluorography showed multiple polypoid lesions, mainly in the jejunum. Ultrasonography, computed tomography, and magnetic resonance imaging showed diffuse cystic tumors in the mesentery and spleen. Cystic lymphangiomatosis was proved by histologic findings of the biopsied specimen at laparotomy. PMID- 9412930 TI - Malignancy in Peutz-Jeghers syndrome? The pitfall of pseudo-invasion. AB - Peutz-Jeghers syndrome is a malignancy-associated polyposis syndrome. We describe a histopathologic phenomenon easily encountered when examining the nature of Peutz-Jeghers polyps but that is underreported in the literature. This phenomenon of "pseudo-invasion" may mimic invasive carcinoma due to epithelial displacement and erroneously give the impression, both macroscopically and microscopically, that a malignancy is involved. This potential pitfall is illustrated by the case of a patient with Peutz-Jeghers syndrome who was thought to harbor a metastasizing adenocarcinoma in his small bowel with peritoneal metastasis as a perioperative finding. Histologic examination, however, revealed pseudo-invasion. PMID- 9412932 TI - Esophageal bezoar in achalasia: a rare condition. PMID- 9412931 TI - Helicobacter pylori infection and atrophic gastritis. A case-control study in a rural town of Japan. AB - We enrolled five hundred twenty-one patients with atrophic gastritis and 769 controls in a case-control study of atrophic gastritis and Helicobacter pylori infection and lifestyle factors. The participants had no history of upper gastrointestinal surgery. They were selected from 1,395 adult (35 years or older) residents of a rural town in Kyoto prefecture who participated in an annual health examination in 1987. Atrophic gastritis was diagnosed on the basis of serum pepsinogen levels: pepsinogen I levels (< or = 70 ng/ml) and pepsinogen I/pepsinogen II ratios (< or = 3.0). The presence of immunoglobulin G antibodies to H. pylori indicated H. pylori infection. At the time of the health examination, participants were interviewed about daily lifestyle habits such as use of cigarettes and alcoholic beverages and consumption of green-yellow vegetables. Unconditional logistic regression analysis indicated that H. pylori infection was associated with a significantly increased risk for atrophic gastritis for all participants (odds ratio, 8.17; 95% confidence interval, 5.63 11.84); for men (odds ratio, 10.91; 95% CI, 5.25-22.67); and for women (odds ratio, 7.28; 95% CI, 4.72-11.22). We found no statistically significant relations between lifestyle factors and atrophic gastritis. PMID- 9412933 TI - Ileitis caused by Henoch-Schonlein purpura. An endoscopic view of the terminal ileum. PMID- 9412934 TI - Colitis associated with alpha interferon? PMID- 9412935 TI - Successful treatment of fissure in ano using topical nitroglycerin. PMID- 9412936 TI - Pancreatic adenocarcinoma on the dorsal part of the pancreas divisum presenting as acute recurrent pancreatitis. PMID- 9412937 TI - Massive volume paracentesis (up to 41 liters) for the outpatient management of ascites. PMID- 9412938 TI - Long delay before celiac disease is recognized. PMID- 9412939 TI - The scratch test is unreliable for detecting the liver edge. AB - To determine the validity and reliability of the scratch test method for estimating the liver span below the right costal margin, we performed a prospective double-blind study using multiple examiners at different levels of training. Twenty-two patients were examined by 11 observers using only the scratch test. Measures of liver edge length below the right costal margin using the scratch test were compared with those by ultrasound. The validity of the scratch test was determined by simple linear regression and the concordance correlation coefficient. There was very poor correlation between the scratch test estimates and ultrasound measurements for all examiners. Measurement reliability, estimated using generalizability theory, for seven observers who examined the same nine patients was 0.68. The reliability of a single examiner was 0.24. The scratch test method for measuring the liver edge span below the right costal margin was neither a valid nor reliable method of physical examination. PMID- 9412940 TI - Urease-based tests for Helicobacter pylori gastritis. Accurate for diagnosis but poor correlation with disease severity. AB - To determine whether the urease-based CLOtest or low-dose 14C urea breath test can predict severity of gastritis or the presence of peptic ulcers, we studied 84 patients presenting for upper endoscopy. Antral biopsies were obtained for histologic analysis and CLOtesting, and urea breath testing was performed. The time to a positive CLOtest and the breath test peak values were correlated with endoscopic findings, severity of gastritis, and bacterial burden. Twenty-four patients had positive urea breath test results (22 with positive CLOtests). Patients with positive breath test results were more likely to have duodenal ulcers, higher grades of gastritis, and increased bacterial burden (p < 0.01 for all comparisons). Correlation between the time to a positive CLOtest or peak breath test values and gastritis severity or bacterial burden was poor (p > 0.05 for all comparisons). In patients with positive tests, no difference was observed between the time to a positive CLOtest or peak breath test value in patients with or without peptic ulcers (p < 0.2 for all comparisons). The low-dose 14C urea breath test and the CLOtest are both accurate for Helicobacter pylori diagnosis. However, neither test predicts the severity of the gastritis, the degree of bacterial burden, or the presence of peptic ulcers. PMID- 9412941 TI - Celiac sprue and immunodeficiency states: a 25-year review. AB - Immunoglobulin deficiency, especially deficiency of IgA, has been described in patients with celiac sprue (CS). Our study was performed in an area of high prevalence of CS to determine the prevalence of immunodeficiency states in patients with CS, to examine their clinical characteristics, response to treatment, and HLA phenotypes compared with a group of age- and sex-matched persons with CS but without immunoglobulin deficiency. Fourteen of 604 patients with CS were identified as being selectively deficient in IgA, whereas one had common variable immunodeficiency. At diagnosis, anemia was present in 8 of 14 IgA deficient patients compared with 10 of 42 controls (p = 0.047), whereas abdominal pain was more common in controls with CS. Autoimmunity and recurrent infection were more prevalent in the IgA-deficient group. Response to gluten-free diet was similar in both groups in terms of histologic structure and recovery of intestinal brush-border enzyme activity. IgA-deficient participants with CS had no increased risk of associated malignancy or lymphoma. HLA phenotypes were similar in both groups. The prevalences of selective IgA deficiency and common variable immunodeficiency in this series of patients with CS are 2.31 in 100 and 0.16 in 100, respectively. Although this group is unique in character, close follow-up coupled with conscientious compliance with a gluten-free diet, remains the mainstay of treatment for these patients. PMID- 9412943 TI - Abnormal common bile duct sonography. The best predictor of choledocholithiasis before laparoscopic cholecystectomy. AB - We conducted a prospective study to determine the most reliable indicator of common bile duct stones before laparoscopic cholecystectomy. One hundred thirty seven patients were referred for endoscopic retrograde cholangiography before laparoscopic cholecystectomy for suspected choledocholithiasis due to one or more of the following abnormalities: (a) altered liver function tests, (b) increased serum amylase levels, or (c) a dilated common bile duct (> 7 mm) with or without evidence of stones on sonography. Sensitivity, specificity, positive and negative predictive values, and the likelihood of the presence or absence of morbidity were calculated for 25 different variables. Common bile duct stones were detected in 63 (46%) patients. Abnormal result of sonography of the common bile duct was the best predictor of choledocholithiasis (p < 0.0001). Abnormalities of the combined liver function tests were statistically significant predictors only when combined with abnormal sonographic results. Improving liver function tests before endoscopy had a significant negative predictive value (p = 0.01). Stepwise logistic regression analysis showed that abnormal ultrasound and the presence of common bile duct stones on ultrasound were significant (p = 0.009 and p = 0.049, respectively). Abnormal result of sonography of the common bile duct is the best predictor of choledocholithiasis before laparoscopic cholecystectomy. PMID- 9412942 TI - Intestinal protein leakage in the acquired immunodeficiency syndrome. AB - Body wasting, protein catabolism, and hypoalbuminemia are complicating features of the acquired immunodeficiency syndrome (AIDS). Given their multifactorial causes, the contributing role of intestinal protein loss has not yet been fully elucidated. To quantify enteric protein leakage, determination of fecal alpha 1 antitrypsin (AAT) excretion has been established as an accurate and reliable endogenous marker. We estimated AAT concentration by standard immune nephelometry in duplicate random stool samples of 49 patients with AIDS, and we compared it to that of 43 patients with chronic inflammatory bowel disease and to 34 healthy controls. When compared with healthy persons, patients with AIDS had increased fecal AAT excretion regardless of current opportunistic intestinal infections and fecal AAT excretion similar to that of patients with quiescent chronic inflammatory bowel disease. The ratio of fecal and serum AAT concentration was not different between AIDS patients and healthy controls, although it was consistently increased in those with chronic inflammatory bowel disease. Significant intestinal protein leakage occurs in patients with AIDS, probably due to primary impairment of gut permeability. Enteric protein loss may be an important feature of human immunodeficiency virus-associated enteropathy with altered mucosal barrier function. PMID- 9412944 TI - Pravastatin has no effect on bile lipid composition, nucleation time, and gallbladder motility in persons with normal levels of cholesterol. AB - Pravastatin dissolves gallstones in patients with hypercholesterolemia by reducing the cholesterol saturation index (CSI) of bile. There are few reports on effect of pravastatin on bile lipids, CSI and nucleation time (NT) in patients with gallstones and normal plasma lipid levels, or on the effect of pravastatin on gallbladder motility. Therefore we studied the effect of pravastatin on bile lipids, CSI, NT, and gallbladder motility in persons with normal cholesterol levels. We included 10 patients (ages 32 +/- 8 years; 6 men) with symptomatic gallstones and normal plasma lipid profiles. Estimation of bile lipids, CSI, and NT in duodenal bile and gallbladder motility were done using standard methods. Subsequently each patient was given 40 mg pravastatin daily for 1 month. At completion of pravastatin therapy, bile lipids and gallbladder motility studies were repeated. After pravastatin therapy, we found no significant reduction in bile cholesterol (11.2 +/- 3.2 vs. 10.4 +/- 2.8 mmol/l), bile acids (114.6 +/- 7.4 vs. 133 +/- 16 mmol/l), phospholipids (23 +/- 3.5 vs. 24 +/- 6.2 mmol/l), CSI (1.28 +/- 0.4 vs. 1.22 +/- 0.3), and nucleation time (7 +/- 3 vs. 7 +/- 3 days). In addition, there was no significant change in gallbladder fasting volume (26 +/ 3 vs. 26.6 +/- 3 ml), residual volume (14.6 +/- 1.1 vs. 15.08 +/- 1.4 ml), ejection fraction (44% vs. 43%), and rate constant of gallbladder emptying (0.018/min vs. 0.022/min). One-month therapy with pravastatin does not alter bile lipids, CSI, NT, and gallbladder contractility in persons with normal levels of cholesterol. PMID- 9412945 TI - Hepatitis C virus antibodies in systemic lupus erythematosus. AB - To determine the prevalence and significance of serum antibody to hepatitis C virus (HCV) in patients with systemic lupus erythematosus (SLE), we measured serum antibodies to HCV by enzyme-linked immunosorbent (ELISA) and by Abbott MATRIX Immunoblot assays in 42 patients with SLE, a condition associated with hypergammaglobulinemia. We used the polymerase chain reaction (PCR) to identify patients with HCV viremia. Five of 42 (11.9%) patients were seropositive for anti HCV by ELISA; of these only two were positive by PCR; only one of three patients seropositive by Immunoblot assay was also positive by PCR. Both ELISA and the Immunoblot assays may be falsely positive for ongoing HCV infection in patients with SLE. Suspected HCV infection should be confirmed with PCR for serum HCV RNA in these patients. PMID- 9412946 TI - Response to interferon-alpha 2a in patients with e antigen-negative chronic hepatitis B. AB - Sixty-eight consecutive patients with chronic hepatitis B received 702 million units of recombinant interferon-alpha 2a. Of the 24 patients negative for hepatitis B e antigen (HBeAg) in serum, the normalization of serum transaminase occurred in 14 (58%) at the completion of interferon therapy and in 13 (54%) at 12 months thereafter; it was normalized in 17 (39%) and 13 (30%), respectively, of the 44 HBeAg-positive patients. Of the HBeAg-negative patients, hepatitis B virus DNA was cleared from serum in six (25%) at the completion and in one (4%) at 12 months thereafter, in contrast to only one (2%, p < 0.05) and none of the HBeAg-positive patients, respectively. The 1896th nucleotide of G (G1896) for codon 28 for tryptophan or A (A1896) for the stop codon 28 in the precore region was determined by restriction fragment length polymorphism. The ten HBeAg negative patients with A1896 only in the precore region had lower pretreatment levels of viral markers, which decreased more rapidly and extensively after interferon than in the 14 HBeAg-negative patients with a mixture of G1896 and A1896 or in the 44 HBeAg-positive patients. These results indicate that patients with HBeAg-negative chronic hepatitis B may respond better to interferon than HBeAg-positive patients, and that the precore mutant with the stop codon 28 may be sensitive to interferon. PMID- 9412947 TI - Strip biopsy to treat esophageal granular cell tumor. AB - Esophageal granular cell tumors are rare neoplasms. We successfully treated a 35 year-old Japanese man with an esophageal granular cell tumor without any complications using strip biopsy. Endoscopic ultrasonography revealed a hypoechoic tumor with a diameter of 8 mm that was confined to the submucosal layer. A strip biopsy done with a two-channel endoscope completely resected the tumor. Six months later, no abnormal findings were recognized in the resected area. Therefore we propose that strip biopsy be considered as a viable alternative treatment for esophageal granular cell tumor, depending on the histologic character, tumor size, and depth of tumor infiltration. PMID- 9412948 TI - Omeprazole-induced interstitial nephritis. AB - Acute renal impairment secondary to interstitial nephritis is a rare complication of omeprazole. We describe a 50-year-old woman who took 20 mg omeprazole twice daily for endoscopically proved ulcerative esophagitis. At the same time, Duke's C colonic cancer was diagnosed and completely resected. Five fluorouracil/folinic acid adjuvant chemotherapy was tolerated without diarrhea or mouth ulceration. Renal function was normal before her first monthly cycle but markedly deteriorated immediately before the second cycle was due. The patient was symptomatic with lethargy, nausea, and mild vomiting, but she was clinically normotensive and only mildly dehydrated. Her serum creatinine concentration increased despite prolonged intravenous hydration, peaking at 4.4 mg/dl 1 week later. Results of a renal ultrasound were normal, and urinary microscopic findings were unremarkable. Renal biopsy showed interstitial nephritis, and renal function improved on cessation of omeprazole, eventually returning to normal. We describe the 12 cases of omeprazole-induced interstitial nephritis reported previously. PMID- 9412949 TI - Retroperitoneal perforation in ulcerative colitis with mediastinal and subcutaneous emphysema. AB - Retroperitoneal colonic perforation in patients with ulcerative colitis is rare. We report such a case in a patient with severe ulcerative colitis without toxic dilatation in whom mediastinal and subcutaneous emphysema also developed. Unlike previously reported cases, our patient was treated conservatively with intravenous fluids, parenteral nutrition, intravenous hydrocortisone, and antibiotics. After 2 weeks, the mediastinal and subcutaneous emphysema and the retroperitoneal air completely disappeared. PMID- 9412950 TI - Human immunodeficiency virus infection and Crohn's disease: the role of the CD4 cell in inflammatory bowel disease. AB - Crohn's disease is believed to have an immunologic basis. The importance of the CD4 cell in particular has been supported by several reports of patients whose symptoms of Crohn's disease resolved after a decline in CD4 count associated with human immunodeficiency virus (HIV) infection. A patient with known Crohn's disease, however, who was later infected with HIV, was reported to continue to have symptomatic Crohn's disease despite an eventual decrease in CD4 count to 84/mm3. We report the new onset of Crohn's disease in an HIV-infected patient with a CD4 count of 100/mm3. This report is the first to document the new onset of Crohn's disease in a patient with HIV and a CD4 count in the range commonly associated with various opportunistic infections and neoplasms. In addition, it is the first to confirm the recent finding that Crohn's disease may be active despite the profound immune deficiency associated with advanced HIV infection. Thus this report further challenges the significance of the CD4 cell in the pathogenesis of Crohn's disease. PMID- 9412951 TI - Rectal variceal bleeding treated by transjugular intrahepatic portosystemic shunt. Potentials and pitfalls. AB - Bleeding from anorectal varices can be massive and life threatening. Prompt differentiation between hemorrhoids and anorectal varices is crucial in treating these patients. Many different treatments are available for bleeding anorectal varices, but none has proved efficacy. We report a case of successful transjugular intrahepatic portosystemic shunt (TIPS) in controlling massive rectal variceal bleeding in an elderly patient with primary biliary cirrhosis and portal hypertension. After TIPS, rapid decompensation of liver function and encephalopathy developed and led to her death. Although TIPS may be effective in controlling acute life-threatening bleeding from anorectal varices, it can be associated with life-threatening complications. PMID- 9412952 TI - Severe necrotizing pancreatitis caused by organophosphate poisoning. AB - Pancreatitis secondary to organophosphate insecticide toxicity is rare and is believed to follow a subclinical and uneventful course. We report two cases of severe acute pancreatitis complicated by pancreatic necrosis and retroperitoneal sepsis in which the diagnosis was obscured by the systemic effects of organophosphate toxicity. Awareness of this complication should prompt earlier investigation because early diagnosis coupled with timely therapeutic measures may improve patient prognosis. PMID- 9412953 TI - Spinal extramedullary hematopoiesis secondary to hepatocellular carcinoma. Case report and literature review. AB - A 38-year-old man, with hepatitis C and a history of intravenous drug use had hepatocellular carcinoma. Severe low back pain was the result not of metastases but of extramedullary hematopoiesis. This was not confirmed by biopsy but seemed unequivocal on magnetic resonance imaging. PMID- 9412954 TI - The role of Peyer's patches in the age-related incidence of Crohn's disease. AB - Recently researchers have suggested that clinical subsets of Crohn's disease occur, which are variously described as inflammatory, fibrostenotic, and fistulizing. In addition, it has been observed that within families with multiple cases, often there is concordance of the site and type of disease. The lesions of Crohn's disease occur in segments that suggest that distribution of Peyer's patches. When the age-related incidence of Crohn's disease was plotted for all countries from which such data were available, the peaks of greatest case frequency occurred at ages 15 to 25 years and paralleled a similar peak representing the number of Peyer's patches as a function of age. This correlation suggests that Crohn's disease may develop as an inflammatory process specifically targeting these important lymphoid structures. Similar peaks of activity in the adolescent to early adult years occur for appendicitis and tonsillitis. PMID- 9412955 TI - Colonic disease associated with a positive assay for Clostridium difficile toxin: a retrospective study. AB - In this retrospective review of colonic tissue from 21 patients with a positive stool assay for Clostridium difficile toxin, four groups of patients were identified by pathologic examination. Classic pseudomembranous colitis was identified in 38% of patients in colon biopsies, resections, and at postmortem examination. One third of patients had acute colitis without specific features on colon biopsies at the time of a positive toxin assay. Effects of C. difficile toxin in patients with idiopathic inflammatory bowel disease (10%) could not be pathologically separated from activity of the underlying disease. In 19% of patients, no acute or chronic colitis or pseudomembranous colitis was noted. This report reminds gastroenterologists that C. difficile infection is associated with a range of pathologic changes similar to the well known clinical spectrum of disease. PMID- 9412956 TI - Significance of short-segment Barrett's esophagus. AB - Barrett's esophagus can progress to dysplasia and adenocarcinoma. Although the incidence of adenocarcinoma of the gastroesophageal junction has increased suddenly in the United States and Europe, we do not know how much of this increase is related to Barrett's esophagus. Interest in mucosal cell abnormalities at the gastroesophageal junction has led researchers to re-examine short-segment Barrett's esophagus. In this recently described condition, specialized columnar epithelium is found in the distal 2 to 3 cm of the esophagus, yet it is not clear how it relates to conventional long-segment Barrett's esophagus, in which the metaplastic epithelium extends higher than 2 to 3 cm above the squamocolumnar junction. The reported prevalence of short-segment Barrett's esophagus found on diagnostic endoscopy varies from 8% to 32%. This wide variation would be lessened by standardized location of biopsy specimens and of endoscopic and histologic staining techniques. Based on the information available, it is apparent that the age range and sex ratios are similar. Although reflux symptoms may be more common in short-segment Barrett's esophagus, disturbances in esophageal motility are less severe and there is less reflux as measured by continuous pH monitoring. Furthermore, recognized complications of Barrett's esophagus, such as ulceration, stricture, high-grade dysplasia, and adenocarcinoma, appear to be uncommon in short-segment Barrett's esophagus. PMID- 9412957 TI - Pancreatic cystadenocarcinoma in Peutz-Jeghers syndrome. PMID- 9412958 TI - Tubular adenoma of the appendix diagnosed before operation. PMID- 9412959 TI - Ulcerative colitis associated with autoimmune progesterone dermatitis. PMID- 9412960 TI - Cholangitis secondary to choledochal cyst in pregnancy and puerperium. PMID- 9412961 TI - Overt polycythemia vera after splenopneumopexy in a patient with Budd-Chiari syndrome. PMID- 9412962 TI - Calendar fluctuations in duodenal ulcer. PMID- 9412963 TI - The role of endoscopy in the diagnosis and follow-up of low-grade gastric mucosa associated lymphoid tissue lymphoma. PMID- 9412964 TI - Changes in acid secretion over the years. A 30-year longitudinal study. AB - We studied the effect of aging on gastric acid secretion in 11 physicians who had augmented histamine tests while at medical school in 1962. One of them had a duodenal ulcer at the time. The augmented histamine test was repeated in 1991 and, in addition, upper gastrointestinal endoscopy was done to exclude peptic ulcer and to obtain biopsies for histologic analysis and assessment of Helicobacter pylori status. The mean basal acid output decreased from 7.3 to 1.9 mEq/hr during the 30-year period of follow-up (p < 0.001), and the mean maximum acid output decreased from 29.9 to 20.3 mEq/hr (p < 0.01). The maximum acid output data showed a profound decrease in 4 of the 11 participants, a lesser decrease in 4, and a minimal increase in the remaining 3. Histologic analysis suggested a greater likelihood of atrophic gastritis, H. pylori infection, or both in participants showing a pronounced decrease in acid secretion with aging. PMID- 9412965 TI - Effect of smoking on the results of esophageal pH measurement in clinical routine. AB - Because data on the effects of smoking on gastroesophageal reflux are controversial, we evaluated the effect of smoking on the results of esophageal 24 hour pH-metry in clinical routine. Participants were 280 consecutive patients with symptoms suggestive of reflux disease, 78 smokers, and 202 nonsmokers. Of the smokers, 45 actually smoked during the pH measurement and 33 abstained from smoking. The frequency of reflux episodes, the fraction of time pH was < 4, and the percentage of abnormal 24-hour pH-metry results were compared among actual smokers, abstaining smokers, and nonsmokers. In actual smokers, the effect of smoking on gastroesophageal reflux was further analyzed by comparing the reflux frequency and the fraction of time that pH was < 4 for a 10-minute period before, during, and after smoking. We found no difference in reflux frequency and fraction of time that pH was < 4 among actual smokers, abstaining smokers, and nonsmokers, regardless of a normal or an abnormal pH-metry result. The percentage of patients with a pH-metry result indicating disease was similar in the three groups, at 53%, 52%, and 50%, respectively. Gastroesophageal reflux was not increased during smoking a cigarette or in the postsmoking period compared with the presmoking period. Neither being a smoker nor actually smoking a cigarette had a negative influence on gastroesophageal reflux. Thus smoking or abstaining from smoking does not modify the results of pH-metry in clinical routine. PMID- 9412966 TI - Typhoid fever and asymptomatic human immunodeficiency virus infection. A report of 10 cases. AB - Ten patients with asymptomatic human immunodeficiency virus (HIV) infection were treated for typhoid fever at King Edward VIII Hospital, Durban, South Africa, from 1993 through 1995. The mean age was 23.7 years (range, 8-33 years), with a female-to-male ratio of 9 to 1 and mortality and morbidity rates of 20% and 10%, respectively. Common presenting manifestations were fever (100%), relative bradycardia (50%), and diarrhea (40%). With respect to epidemiologic and clinical characteristics, we noted no significant differences among these 10 HIV-positive and 32 HIV-negative patients treated for typhoid fever during the same period. However, we found hepatic dysfunction in the form of an isolated increase in aspartate aminotransferase (p < 0.01) and abnormal urinary findings suggestive of glomerulonephritis (p = 0.01) more frequently in HIV-positive patients. PMID- 9412967 TI - Fecal incontinence in scleroderma. Clinical features, anorectal manometric findings, and their therapeutic implications. AB - Fecal incontinence is an under-reported complication of scleroderma. Ten incontinent patients with scleroderma were evaluated through anorectal manometry and compared with 20 incontinent patients without scleroderma who were matched for age and sex as controls. The scleroderma patients had a higher voluntary external anal squeeze pressure, whereas the resting internal anal sphincter pressure was similar to that of the control group. The threshold for rectal sensation in the scleroderma group was significantly less than that in controls. Episodes of fecal incontinence, anal canal length, and maximal tolerable volume were not significantly different between the study groups. The rectoanal inhibitory response was abnormal in 80% of patients with systemic sclerosis but was normal in 70% of the controls. Stool consistency was significantly looser in the scleroderma patients. Treatment of fecal incontinence in scleroderma patients may be successful in many patients using a combination of dietary and pharmacologic manipulation because diarrhea is an important etiologic cofactor superimposed on reduced internal anal sphincter pressure. PMID- 9412968 TI - Cholesterolosis is not associated with high cholesterol levels in patients with and without gallstone disease. AB - High levels of cholesterol have been associated with certain gallbladder disorders such as cholesterolosis and gallstone disease. Furthermore, obesity is considered the main risk factor for cholesterol gallstone disease. We investigated the incidence of cholesterolosis in patients with and patients without gallbladder stones (GS). We reviewed the clinical records of patients with gallstone disease and other gallbladder disorders who had consecutive cholecystectomy during a 5-year period. We recorded demographic data, sex, age, serum cholesterol levels, and body mass index. The diagnosis of cholesterolosis was made macroscopically and microscopically. A total of 636 patients were included in this study: 446 with and 190 without GS. Cholesterolosis was more frequent in patients without GS (p < 0.01). However, hypercholesterolemia occurred more frequently in patients with GS (p < 0.001). Obese patients with GS had higher percentages of cholesterolosis and hypercholesterolemia than did eutrophic patients (p < 0.01 and p < 0.05, respectively). We suggest that cholesterolosis in the human gallbladder is not necessarily associated with gallstone disease and high plasma cholesterol levels. PMID- 9412969 TI - Autoimmune disease is not a feature of hepatitis C infection in Ireland. AB - To determine the prevalence of autoimmune disease, autoantibody positivity, or both in Irish persons with hepatitis C, we surveyed 98 such patients (55 recipients of anti-D, 25 intravenous drug abusers, and 18 blood transfusion recipients). We studied them clinically and tested for anti-nuclear, anti-smooth muscle, and anti-mitochondrial, liver-kidney microsomal, thyroid microsomal, thyroid globulin, and gastric parietal antibodies; and also for rheumatoid factor. In the anti-D antibody group (all female), two patients reported generalized musculoskeletal symptoms but had no demonstrable physical signs. We did not find cryoglobulins in any patient. We detected thyroid microsomal antibodies in only 6 of 55 (10.9%) patients. (In two of these, thyroid globulin antibodies were also positive). These patients were all clinically euthyroid, but two had borderline low-normal thyroid function tests. Titers for anti-nuclear antibodies were weakly positive in 5 of 55 (9.1%) patients, and gastric parietal antibodies were positive in 5 of 55 (9.1%) patients. In particular, we noted no antibodies to liver-kidney microsome. Rheumatoid factor was detected in eight patients. Forty-seven of 55 patients were genotype 1b, and 8 of 55 were genotype 3. In the intravenous drug abusers (8 women, 17 men), we detected no autoantibodies. Seven of the 25 genotypes were tested; three were genotype 3 and four were genotype 1b. In the transfusion group (10 women, 8 men), we detected no autoantibodies apart from weak anti-nuclear antibody Titers (1:10), which we found three patients. Five of 10 genotypes tested were of genotype 3 and the other five were of genotype 1b. These findings suggest that in Irish patients with hepatitis C, neither genotype nor source (and dose) of inoculum contributes to the development of autoimmune disease. How hepatitis C virus is associated with autoimmune disease in other studies remains unknown. The answer may, at least in part, be found in genetic; HLA typing studies should provide useful information. PMID- 9412970 TI - Utility of hepatitis C virus RNA levels for predicting the therapeutic efficacy of interferon. AB - We studied the levels of serum hepatitis C virus (HCV)-RNA, the HCV genotype before interferon therapy, and the kinetics of serum HCV-RNA at the initial stages of therapy to determine their utility in predicting the therapeutic efficacy of interferon in 44 patients with chronic hepatitis C infection. We also looked at the efficacy of repeated interferon treatment in relation to the kinetics of serum HCV-RNA. The level of serum HCV-RNA determined by a branched DNA probe assay before interferon treatment and that by a reverse transcription nested polymerase chain reaction assay during the initial stages of interferon administration were useful for predicting the efficacy of treatment. Furthermore, detection of serum HCV-RNA by the reverse transcription nested polymerase chain reaction assay after the completion of interferon therapy indicated relapse at its earliest stage. In patients who experience relapse, repeated treatment with an appropriate dose of interferon before an increase in viral levels may increase the proportion of complete responses. PMID- 9412971 TI - Hepatitis C virus-related liver damage is related to cold activation of complement. AB - A high positivity of cold activation of complement has been reported in Japanese patients having hepatitis B virus-negative chronic hepatitis. Although the cause of cold activation of complement is unknown, the involvement of hepatitis C virus (HCV) has been suspected. We studied the positivity of cold activation of complement in 253 patients, including 93 patients with chronic hepatitis C infection who received 6MU natural interferon-alpha per day for 24 continuous weeks. Cold activation was positive in 38% of patients with chronic hepatitis C and in 46% of patients with liver cirrhosis C. We did not detect cold activation in asymptomatic HCV carriers; patients with chronic hepatitis B, liver cirrhosis B, or alcohol-related liver damage; or in the controls. Cold activation was also negative in HCV-RNA-negative patients who responded completely to interferon alpha, and in HCV-RNA-positive patients who responded partially whose serum alanine transaminase levels were normalized after interferon treatment. In the patients who had a relapse of hepatitis C after interferon treatment, positivity of cold activation increased sharply. We conclude that HCV-associated liver damage is related to the development of cold activation of complement. Cold activation is useful for monitoring the response to interferon in patients with chronic hepatitis C infection. PMID- 9412972 TI - Polyarteritis nodosa associated with gastric carcinoma and hepatitis B virus infection. AB - Several cases of polyarteritis nodosa associated with malignant disorders have been reported, most with bone marrow-related tumors. We report polyarteritis nodosa presenting with a fever of unknown origin and muscle weakness that was complicated by advanced gastric carcinoma and hepatitis B virus-positive cirrhosis. Vasculitis was diagnosed after gastrectomy from histologic findings of arterial vasculitis on the resected gastric carcinoma. Our case is so far the second such report of polyarteritis nodosa associated with gastric cancer. PMID- 9412973 TI - Nonalcoholic steatohepatitis masquerading as autoimmune hepatitis. AB - Nonalcoholic steatohepatitis is an increasingly recognized clinicopathologic condition. We report two cases of nonalcoholic steatohepatitis in middle-aged Japanese women whose clinical and laboratory data mimicked autoimmune hepatitis. Histologic findings of both cases were definite steatohepatitis with portal and pericellular fibrosis. Both patients' HLA-DR haplotypes were DR4 and DR2, which are frequently observed in Japanese patients with autoimmune hepatitis. Our cases suggest a diversity in the pathogenesis of nonalcoholic steatohepatitis. PMID- 9412974 TI - Abdominal tuberculosis involving hepatic hilar lymph nodes. A cause of portal vein thrombosis and portal hypertension. AB - Abdominal tuberculosis is a rare disease. Involvement of lymph nodes at the hepatic hilum responsible for jaundice is exceptional. We describe a 59-year-old woman in whom jaundice was the presenting feature, complicated by portal vein thrombosis and portal hypertension. PMID- 9412975 TI - Human monocytic ehrlichiosis presenting as febrile diarrhea. AB - I report a case of tick-borne ehrlichiosis in which the primary presentation suggested an infectious diarrhea, with impressive leukopenia and thrombocytopenia. Ehrlichiosis must be included in the differential diagnosis of febrile diarrhea in endemic areas because delay in therapy allows complications and sometimes death. In patients with apparent febrile diarrhea, the presence of leukopenia and thrombocytopenia or elevated aminotransferase values should alert the physician to look for a history of tick bites and consider treatment with doxycycline or tetracycline. PMID- 9412976 TI - Histopathologic study of islets of Langerhans in patients after gastrectomy. AB - In autopsy specimens removed after gastrectomy from patients who had survived for 2.5 to 40 years after surgery, we found hyalinization of the islets of Langerhans in 9 of 15 cases (60%) with Billroth's I anastomosis (B-I) and in 8 of 16 cases (50%) with Billroth's II anastomosis (B-II). The hyalinization in the postgastrectomy patients was significantly increased compared with that in controls (p < 0.01). The percentage of hyalinized islets of Langerhans in the 17 cases showing hyalinization ranged from 0.2% to 14.3%. The percentage increased relative to the period after gastrectomy in the B-I patients but not in those with B-II. The blood glucose level was mildly elevated in three B-I patients, whereas it was increased in only one patient with B-II. Hyalinization of the islets of Langerhans may be due to vagotomy or to excessive stimulus and hyperactivation of the islets. In any event, the islets of Langerhans in patients after gastrectomy were frequently replaced by hyalinization, which might cause impaired glucose tolerance. PMID- 9412977 TI - Primary coexistent adenocarcinoma and choriocarcinoma of the stomach. A case report and review of the literature. AB - We report a case of primary gastric choriocarcinoma with liver metastasis. The mixed histologic patterns included adenocarcinoma, undifferentiated carcinoma, and choriocarcinoma. Immunohistologic staining for the beta-subunit of human chorionic gonadotrophin (beta-HCG) showed positive results in the choriocarcinoma, adenocarcinoma, and normal mucosal gland. However, positive HCG cells were present at different intensities in the choriocarcinoma, adenocarcinoma, and normal mucosal gland. The level of HCG was significantly increased in serum. This unusual tumor probably resulted from dedifferentiation of a primary adenocarcinoma or developed directly from the mucosal glands. PMID- 9412978 TI - Colonic ischemic stricture presenting as a late complication of the hemolytic uremic syndrome. PMID- 9412979 TI - Beclomethasone through artificial cecocutaneous fistula for ulcerative colitis: a case report. PMID- 9412980 TI - Osseous metaplasia in benign rectal polyps. PMID- 9412981 TI - Adverse pregnancy outcome in the antiphospholipid syndrome: focus for future research. AB - Pregnant patients with antiphospholipid syndrome (APS) may suffer from recurrent pregnancy loss, pre-eclampsia, intrauterine growth restriction and placental abruption. These conditions inevitably result in a high incidence of premature delivery with all the neonatal complications that follow. The mechanism underlying these adverse pregnancy outcomes has not yet been established. This may be primarily a maternal disease process with secondary placental maldevelopment and/or malfunction. Alternatively, there may be primary placental damage mediated directly or indirectly by antiphospholipid antibodies. The safe and successful treatment of pregnant women with APS lies in the understanding of the aetiology of this condition and the mechanism by which complications in pregnancy may arise. In this article we highlight areas in which research may be targeted such that our understanding of the pathogenesis of adverse pregnancy outcome may be enhanced. PMID- 9412982 TI - Hepatitis C virus, autoimmunity and rheumatic disease. PMID- 9412983 TI - Systemic lupus erythematosus in India. AB - The first case of systemic lupus erythematosus (SLE) was reported from India in 1995 followed by two more case reports and further, a series of eight cases, till 1969. Since the establishment of a clinical immunology laboratory at a major teaching institution in New Delhi in 1968, SLE was extensively studied and reported from that centre. From mid-1980 onwards several other centres in different regions in India including Chennai (old name Madras), Mumbai (old name Bombay), Calcutta and Hydrabad, also published their regional experience on SLE. Based on these data, the present report describes the clinical and laboratory characteristics of 1366 SLE patients seen in different regions of India. Arthritis, rash, photosensitivity, seizures and psychosis were seen in comparable proportions to other racial groups. Similarly, ANA and anti-DNA antibody positivity was also within the range seen in other racial groups. When compared with other series, however, alopecia, renal lupus, oral ulcers and neurological involvement was seen in higher proportions, reaching statistically significant figures in comparison to some racial groups. In contrast, haematological manifestations were seen in significantly less proportions in comparison to some of the racial groups. Serositis and discoid lesions were also seen in lower proportions than in most of other races. The proportion of those with anti-Sm antibodies was in between two extremes of highest among Africans and Israelis and lowest among Chinese and Europeans. Other manifestations were comparable to most other racial groups. Compared to North American and European reports, significantly low 5 and 10 year survival was observed among patients from India. This could be related to the general public health situation in the country including less than optimal management facilities in hospitals, delay in diagnosis due to lack of awareness of the disease, referral bias where only serious patients reach major city hospitals, or a truly severe disease among Indians, or a combination of these genetic, environmental and/or sociocultural factors. The Main causes of death were irreversible renal damage, infections and neurological involvement. Despite a comparable prevalence of anticardiolipin antibodies (aCL) and lupus anticoagulants (LAC), clinical antiphospholipid syndrome was significantly less common. Genetic studies showed appreciable increase of HLA DR4 (37.5%) among patients compared with controls (18%). Additionally the haplotype B8-DR3 was encountered frequently in the patient group. PMID- 9412984 TI - Detection of restricted junctional diversity of peripheral T cells in SLE patients by spectratyping. AB - Analysis of somatic mutations revealed that anti-double-stranded DNA (dsDNA) autoantibodies from patients with systemic lupus erythematosus (SLE) share features of a T cell dependent, antigen driven immune response. Therefore we analysed the length diversity of the complementarity determining region 3 (CDR3) of T cell receptor (TCR) by high resolution gel electrophoresis of 16 V beta family specific RT PCR products (spectratyping). To enable statistical analysis we developed a quantitative scoring method for the histograms. We investigated 16 V beta gene families in peripheral T cells of SLE patients (n = 9) with active (n = 5) and inactive (n = 4) disease as well as normal healthy blood donors (NHD; n = 9). Analysis of TCR V beta spectratypes (active SLE, n = 59; inactive SLE, n = 51 and NHD n = 97) revealed statistically significant differences of CDR3 length distribution between SLE patients and NHD (P < 0.0001 (active SLE/NHD) and P = 0.0034 (inactive SLE/NHD). These results suggest that spectratyping is able to detect clonal activation of peripheral T cells which correlates to disease activity in SLE patients. We conclude that peripheral T cells from SLE patients display features of a secondary antigen driven immune response. PMID- 9412985 TI - Lack of NK cells in lupus patients with renal involvement. AB - We have previously shown that patients with SLE have significantly lower percentages and absolute numbers of NK(CD3-/CD16+56) cells in their peripheral blood compared with normals. Patients with active disease had very low levels of NK cells and the reduction was also associated with patients who had renal involvement. We have now performed a serial study immunophenotyping 11 patients with SLE and renal involvement using dual colour immunofluorescence and flow cytometry. Patients were tested every three months on an average of three occasions. As a control, nine SLE patients without renal involvement were immunophenotyped for similar intervals; 11 normal controls were also tested. Major lymphocyte subsets (T, B and NK) remained very stable during serial bleeds. However, the NK cell populations were decreased significantly in patients with renal involvement both as percentages (5 +/- 6 vs 9 +/- 5, P < 0.0001) and absolute counts (75 +/- 108 vs 109 +/- 52, P < 0.001) in comparison to non-renal patients. Analysis of disease activity using BILAG score showed an inverse correlation between renal system activity and percentage and absolute number of NK cells (P < 0.002 and 0.01, respectively). In this study we have also analysed a CD8 T cell subset which we have not studied before. We have found a significantly increased percentage of CD38+CD8+ T cells(activated CD8 subset) in patients with SLE in comparison to normal controls. We did not find any association with the CD38+CD8+ T cells and disease activity as measured by BILAG or renal involvement. NK cells are important factors in immunity against virus infections and tumour cells. CD38+CD8+T cells are increased in viral infections. We speculate that the lack of NK cells in SLE patients might have an association with increased CD38 expression. PMID- 9412986 TI - Pulmonary haemorrhage in Oriental patients with systemic lupus erythematosus. AB - We reviewed the case records of 10 Oriental patients with systemic lupus erythematosus (SLE) who developed pulmonary haemorrhage (PH) between 1987 and 1996 to determine their clinical presentation and outcome. All the patients had clinical evidence of PH including a sudden onset of dyspnoea, tachycardia, fall in haemoglobin (at least 1.5 gm%) and bilateral diffuse alveolar infiltrates on chest radiographs. At the time of PH, nine patients had a disease duration of 2 years or less and all the patients had clinical and/or laboratory evidence of active lupus disease. Fever and lung crepitations were present in 90% of patients while haemoptysis and chest pain occurred in only three and two patients, respectively. All the patients were treated with high dose intravenous corticosteroids and in addition seven had a combination of pulse methylprednisolone and cyclophosphamide, and four had received plasmapheresis. Four patients died as a result of PH. One patient died of pneumonia three years after recovering from PH while the remaining five had no recurrence of PH after a median follow-up of 22 months. Our study suggests that PH in Oriental lupus patients often occurs early in the disease, rarely presents with haemoptysis and has a high mortality despite aggressive immunosuppressive therapy. PMID- 9412987 TI - Elevated soluble fas production in SLE correlates with HLA status not with disease activity. AB - Evidence from animal models of lupus suggests that disruption of Fas-mediated apoptotic events may play a role in systemic lupus erythematosus (SLE). The recently described secreted from of Fas (sFas) could interfere with apoptotic events by blockading Fas/Fas ligand interactions. We describe elevated secreted Fas protein in sera from 60 patients with SLE compared with controls but neither sFas protein nor sFas mRNA levels correlated with disease activity. At the mRNA level there is strong evidence that individuals with human leucocyte antigens common in SLE patients have a genetic predisposition for increased secreted Fas production. PMID- 9412988 TI - Atherosclerosis in LDL-receptor knockout mice is accelerated by immunization with anticardiolipin antibodies. AB - Atherosclerosis is a process initiated by accumulation of macrophages in distinct areas of endothelial cell damage and uptake of large amounts of lipids. Recently, it has been shown that the immune system plays an active part in the progression of the atherosclerotic plaque although its precise role has not yet been elucidated. Anticardiolipin antibodies (aCL) are generally found in the sera of patients with the antiphospholipid syndrome (APS) and are associated with a prothrombotic state. Several authors have demonstrated that aCL can activate platelets and endothelial cells as well as increase oxidized low density lipoprotein (LDL) uptake by macrophages. In the present study we sought to assess the effect of immunization with aCL (Ab1, leading to the production of mouse aCL Ab3) on the progression of atherosclerosis. Two groups of 8-weeks old female LDL receptor knockout mice (n = 13 per group) were immunized with IgG purified from the serum of an APS patient or with normal human IgG, respectively. The aCL immunized mice developed high titres of 'self' aCL (detected using the standard aCL ELISA) as compared with the normal human IgG immunized mice, whereas no differences were noted between both study groups with respect to the serum lipid levels. The extent of fatty streak formation was significantly higher in the aCL immunized mice in comparison with the human IgG injected mice (mean aortic lesion size of 5308 +/- 471 microns2 vs 1027 +/- 184 microns2, respectively, P < 0.01). The immunohistochemical analysis of the atherosclerotic plaques from both mouse groups did not display differences in cellular composition. The results of the study show that mouse aCL induced by immunization with human aCL from an APS patient enhance atherogenesis in LDL-RKO mice and imply that these antibodies may play a role in atherosclerosis development in patients with the APS. PMID- 9412989 TI - Systemic lupus during pregnancy with refractory alveolar haemorrhage: recovery following termination of pregnancy. AB - A case of refractory pulmonary hemorrhage in a pregnant 22-year-old with systemic lupus is presented. The clinical difficulty of management of pulmonary haemorrhage and lupus flare during pregnancy are discussed. PMID- 9412990 TI - Development of systemic lupus erythematosus in a rheumatoid arthritis patient with anti-ribosomal P protein antibody. AB - We describe a patient with rheumatoid arthritis (RA) whose sera contained a high titre of an antibody targeting cytoplasmic ribosomal P proteins (anti-P). This association preceded by 6 years the development of serological and clinical manifestations of systemic lupus erythematosus (SLE). The clinical significance of anti-P for the diagnosis of SLE is discussed. PMID- 9412991 TI - Chronic hepatitis C virus infection, mixed cryoglobulinaemia, leukocytoclastic vasculitis and antineutrophil cytoplasmic antibodies. PMID- 9412992 TI - Aneugenic activity in human cultured lymphocytes. An overall study with colchicine using the micronucleus assay and fluorescence in situ hybridization techniques. AB - The effects induced by aneugenic agents on chromosome segregation are manifold. The biological relevance of these effects has led to the development of assays specifically detecting aneugens. In this context, the micronucleus (MN) assay in binucleated human lymphocytes along with FISH has been considered a pertinent tool for detecting aneugenic and clastogenic activity. However, the MN assay is insensitive in detecting aneugenic effects other than chromosome loss. By using the aneugenic model compound colchicine and X chromosome centromere-specific FISH, we have shown that besides chromosome loss in binucleated cells, other effects such as MN in mononucleated cells, cells arrested at metaphase, polyploidy and non-disjunction are also consistently induced by aneugenic agents. A chromosome 1 centromeric probe was used simultaneously with X chromosome centromeric labeling in mononucleated cells in order to distinguish polysomy from polyploidy. It is concluded that all these effects should be considered for a comprehensive evaluation of aneugenic activity. PMID- 9412993 TI - Inhibition of potassium dichromate mutagenicity by todralazine. AB - Todralazine, an antihypertensive drug of the hydrazinoph-thalazine group, markedly decreased the mutagenic activity of potassium dichromate in standard bacterial tests. At the highest todralazine dose tested inhibition of potassium dichromate mutagenic activity by approximately 90% in the Ames test and up to 100% (complete) inhibition in the Bacillus subtilis rec- assay was observed. Spectrophotometric analyses proved that todralazine induced reduction of Cr(VI) to Cr(III) and complexation of Cr(III) ions. These spectro-photometric results may be a presumptive explanation of the observed mutagenic activity decrease, as it is known that Cr(III) is poorly transported across cell membranes and therefore is not mutagenic to bacterial cells. We perceive our experiments as an example of attempts which should lead to an effective reduction in chromium genotoxic and carcinogenic activity in exposed individuals. PMID- 9412994 TI - Effect of nucleotide excision repair on hprt gene mutations in rodent cells exposed to DNA ethylating agents. AB - The role of the nucleotide excision repair (NER) pathway in removal of DNA ethylation damage was investigated by means of hprt mutational spectra analysis in the NER-deficient Chinese hamster ovary cell line UV5, which lacks ERCC2/XPD, and in its parental cell line AA8. Both cell lines were exposed to ethyl methanesulfonate (EMS) or N-ethyl-N-nitrosourea (ENU). EMS gave a similar dose dependent increase in hprt mutants in UV5 compared with AA8. In both cell lines EMS-induced mutations in the hprt coding region consisted almost exclusively of GC-->AT transitions, probably due to the direct miscoding lesion O6-ethylguanine. ENU, an agent that in addition to O6-ethylguanine also induces other O-alkylation products, was significantly more mutagenic in UV5 than in AA8. Mutational spectra analysis showed that the proportions of ENU-induced GC-->AT, AT-->TA and AT-->GC base pair changes were similar for both cell lines. ENU-induced DNA lesions that may be involved in GC-->AT transitions are O6-ethylguanine and O2-ethylcytosine, the latter being a chemically stable DNA lesion of which the miscoding properties and repair characteristics are largely unknown. ENU-induced AT-->TA transversions are probably caused by O2-ethylthymine, which mispairs with thymine. In AA8 thymines in ENU-induced AT-->TA transversions were exclusively located in the non transcribed strand of the hprt gene, whereas in UV5 30% of these thymines were found in the transcribed strand. Together, these results indicate that O6 ethylguanine is a poor substrate for NER in rodent cells and that O2 ethylpyrimidines are preferentially removed from the transcribed strand of the hprt gene by NER. PMID- 9412995 TI - Induction of micronuclei and sister chromatid exchange in mouse splenocytes after exposure to the butadiene metabolite 3,4-epoxy-1-butene. AB - 3,4-Epoxy-1-butene (EB) is one of the main metabolites of 1,3 butadiene, a widely used industrial chemical. The mutagenic potential of 1,3 butadiene and its metabolites have been studied in different test systems. In this work the genotoxic effects of EB were studied by estimating micronuclei (MN) and sister chromatid exchange (SCE) frequencies in stimulated mouse splenocytes. Mice were treated in vivo with various doses of EB (24.4, 48.8 and 73.2 mg/kg). The antikinetochore antibody technique (CREST) was also applied to MN in cytokinesis blocked cells to investigate any possible aneugenic effect. Both MN and SCE frequencies increased after EB treatment. The induced MN resulted mainly from acentric fragments but a weak aneugenic effect was found as well. Cytotoxic effects of EB were observed at the highest dose. The above results, in combination with others on the effect of 1,3 butadiene and its metabolites in somatic and germ cells of mouse and rat as well as in somatic human cells, form a part of the information needed for application of the parallelogram approach and extrapolation to human risk. PMID- 9412996 TI - Modulation of the potency of promutagens and direct acting mutagens in bacteria by inhibitors of the multidrug resistance mechanism. AB - Multiple drug resistance (MDR) mechanisms are known to limit the effectiveness of some cancer chemotherapies, probably through enhancing P-glycoprotein-mediated drug efflux from mammalian cells. Similar mechanisms appear to act in other organisms, including bacteria, and may affect not only the toxicity but also the mutagenicity of certain chemicals. At least in some experimental situations, MDR can be overcome through concomitant treatment of the cells with various types of inhibitors. Two MDR inhibitors, verapamil, a calcium channel blocker, and trifluoperazine, a calmodulin inhibitor, were assayed for their ability to modulate the potency of nine mutagens with varying mechanisms of action in various Salmonella typhimurium his- strains. Neither verapamil nor trifluoperazine affected the direct mutagenicity of sodium dichromate and 2 methoxy-6-chloro-9[3-(2-chloroethyl)amino-propyl-amino] dihydrochloride (ICR 191) or the S9-mediated mutagenicity of benzo[a]pyrene and 2-amino-3,4-dimethyl amidazo[4,5-f]quinoline (MeIQ). Both modulators enhanced the direct mutagenicity of doxorubicin. Moreover, trifluoperazine sharply increased the S9-mediated mutagenicity of cyclophosphamide and 2-aminofluorene, while it consistently decreased the mutagenicity of 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2). The contrasting effect towards the aromatic amine 2-aminofluorene and the heterocyclic amine Trp-P-2, representative of important chemical families responsible for the bacterial mutagenicity of cigarette smoke, may explain the observed lack of influence of trifluoperazine on the mutagenicity of a cigarette smoke condensate. These observations extend the known range of chemical types whose mutagenicity can be modulated by inhibitors of MDR and suggest that there may be value in adding MDR inhibitors, especially trifluoperazine, to optimize the detection of mutagenicity by certain types of chemicals in the Salmonella/mammalian microsome mutagenicity test. PMID- 9412997 TI - Chromosomes with high gene density are preferentially repaired in human cells. AB - It is known that DNA repair is heterogeneous in human cells since open chromatin, active genes and their transcribed strands are preferentially repaired. It is thus expected that DNA repair is clustered in chromosomes with high gene density. We have employed a DNA repair inhibitor, cytosine arabinoside (Ara-C), to convert ethyl methane sulfonate (EMS)-induced excision repairable lesions to chromosomal breaks, to check for the existence of heterogeneity of repair at the chromosome level. Chromosome staining by fluorescence in situ hybridization (FISH) was used to analyze breakage in chromosomes with diverse gene densities. These chromosomes were identified by means of the CpG island distribution after FISH with a CpG island-rich probe isolated from total human genomic DNA. Thus, three chromosomes with very high gene density (numbers 1, 19 and 20) were compared with two chromosomes with very low gene density (numbers 4 and 18) for clastogenicity and sensitivity to co-treatment with Ara-C and EMS. Our data indicate that those chromosome with higher gene density are more sensitive to a combination treatment with Ara-C and EMS, indicating that the level of excision repair synthesis is higher in those chromosome. It is therefore concluded that DNA excision repair is preferentially directed to chromosomes with high gene density. The implications of this finding in human biomonitoring using FISH techniques are discussed. PMID- 9412998 TI - Photochemical production of uracil quantified in bromodeoxyuridine-substituted SV40 DNA by uracil DNA glycosylase and a lysyl-tyrosyl-lysine tripeptide. AB - Exposure to UVA radiation of SV40 DNA substituted with bromodeoxyuridine (BrdU) in the presence of Hoechst dye 33258 results in the production of uracil. The yield of uracil was determined by measuring the increase in the single-strand break (SSB) yield after incubation of the photolyzed DNA with uracil-DNA glycosylase (UDG) in the presence of the tripeptide lysyl-tyrosyl-lysine (KYK). UDG removes uracil to leave an abasic site which is then cleaved to a SSB by KYK. The SSB yield was quantified by digital video imaging of ethidium fluorescence after separation of the I, II and III forms of SV40 DNA by agarose gel electrophoresis. Uracil is not detected when photolysis is carried out in the absence of the dye nor when unsubstituted DNA is used as the substrate. Without UDG or KYK treatment, the F0 for the loss of form I DNA is 100 J/m2. This falls to 13 J/m2 after incubation with UDG and KYK, indicating that uracil formation is approximately 5-fold more efficient than SSB formation. Formation of uracil suggests a mechanism for the high cellular toxicity of the dye-BrdU-UVA treatment. PMID- 9412999 TI - Radioactive iodine induces clastogenic and age-dependent aneugenic effects in lymphocytes of thyroid cancer patients as revealed by interphase FISH. AB - After the Chernobyl nuclear accident, a dramatic 131I-related increase in the incidence of thyroid cancer has been reported in exposed children. However, little is known about the eventual genotoxic effects of 131I in exposed humans. Thyroid cancer patients are usually treated with 131I and, therefore, they provide us with an opportunity to study cytogenetic damage induced by known doses of this radionuclide. FISH techniques have been employed to study the origin of micronuclei as well as X chromosome non-disjunction and X chromosome numerical abnormalities in lymphocytes from 131I-treated women suffering from thyroid cancer. Blood was sampled before and 1 week after 131I treatment. Cells were analysed with either pancentromeric FISH to classify micronuclei or X chromosome centromere-specific FISH in mononucleated and binucleated cells to evaluate X chromosome numerical abnormalities and non-disjunction respectively. Our data indicate that 131I-induced clastogenic and age-dependent aneugenic effects in the lymphocytes of exposed patients. The X chromosome was not preferentially involved in the aneugenic effect induced by 131I. It is concluded that besides its major clastogenic effect, 131I can also induce an X chromosome-independent aneugenic activity mainly in patients with spontaneous proneness to chromosome loss. PMID- 9413000 TI - Induction of an adaptive response to quercetin, mitomycin C and hydrogen peroxide by low doses of quercetin in V79 Chinese hamster cells. AB - The adaptive response is a phenomenon by which cells exposed to low, non cytotoxic doses of a genotoxicant become significantly resistant to a subsequent higher dose of the same or another genotoxic agent. Induction of the adaptive response has been mainly studied using ionizing radiation and alkylating agents as genotoxic agents. However, other mutagenic agents may warrant further study, since the adaptive response as a whole may be an important general biological mechanism to maintain genetic integrity and thus could prevent carcinogenic initiation of cells. The exposure to mutagenic agents present, or formed, in the diet is considered an important factor in the etiology of human tumors and a considerable number of these agents have not yet been identified or characterized. Flavonoids are a large group of polyphenolic quinoids found in a wide variety of edible fruits and vegetables and a few, such as quercetin, present genotoxic activity in vitro. The mechanisms of mutagenicity of quercetin involve the production of oxygen radicals through an autoxidation process dependent on pH value and the presence of oxygen. Although there are few doubts regarding the mutagenicity of quercetin in vitro, carcinogenicity of flavonoid is still controversial. In view of these conflicting results and the radiomimetic nature of the mutagenicity of flavonoids, we addressed the question of cell exposure to quercetin at the low levels present in the diet leading to adaptation to further exposure to mutagens or carcinogens. The work reported here concerns induction of an adaptive response by low doses of quercetin to challenging doses of quercetin and other compounds, namely hydrogen peroxide and mitomycin C, using induction of chromosomal aberrations in V79 cells as the end point. PMID- 9413001 TI - Effect of 1 Gy X-rays in G1 phase on activation of cell cycle checkpoints in human lymphocytes: role played by chromosomal aberrations. AB - The effect of X-irradiation (1 Gy) during G1 on the transition from G1 to S and on the length of G2 over cell cycles subsequent to irradiation was studied in human lymphocytes from six different donors. After irradiation a delay was observed in the onset of S phase, as was an extension of the G2 phase lasting throughout the three to four subsequent cell divisions. The extension of G2 and of the cell cycle as a whole is partly related to the presence of chromosome aberrations in the cell. This is demonstrated by: (i) the presence of a larger number of chromosome aberrations in M1 cells corresponding to sampling times longer after that of irradiation; (ii) the presence of a larger number of chromosome aberrations in cells with a longer G2. The most significant chromosome aberrations in this respect are isochromatid fragments. Lastly, we observed that irradiation during G1 activates another checkpoint governing the way mitosis proceeds. This takes the form of an extension of metaphase; in this case also, in some cells, activation of a possible checkpoint during preanaphase seems to be related to the presence of aberrations. PMID- 9413002 TI - Tuberous Sclerosis Complex: the role of neuroradiology. AB - Tuberous Sclerosis Complex is the second commonest phakomatosis affecting between 1 in 70,00-10,000 live births. It is a multisystem disorder but neurological disturbances are often the presenting feature and cause major morbidity/mortality. In this paper the neuroradiological and clinical features are reviewed, focusing on a rational approach to imaging children with tuberous Sclerosis Complex. PMID- 9413003 TI - Attenuated auditory event-related potential (mismatch negativity) in children with developmental dysphasia. AB - An attention-independent negative wave-form termed 'mismatch negativity' (MMN) of the auditory event-related potentials (ERPs) was studied in ten children (3-6 years) with developmental dysphasia and in fourteen control children (3-7 years) with normal speech and language development. The MMNs were elicited with pure sine tone stimuli using the oddball paradigm. The peak latency and peak amplitude of MMN response to frequency (500/553 H2) difference was measured. The grand average amplitude of frequency MMN was significantly attenuated in dysphasic children as compared to controls, but no significant difference was observed in the latency of peak frequency MMN. The results indicate that dysphasic children have a defect in automatic auditory processing of frequency differences. Because the MMN response reflects central auditory processing and is modal specific for auditory stimuli we argue that the MMN method can serve as an objective tool to assess central auditory deficits in children. PMID- 9413004 TI - Valproate treatment induces lipid globule accumulation with ultrastructural abnormalities of mitochondria in skeletal muscle. AB - In skeletal muscle of seven children treated with valproic acid (VPA) microvesicular (0.05-2 microns) lipid droplets were observed between the myofibrils predominantly adjacent to the mitochondria. Most of the mitochondria showed ultrastructural abnormalities: several mitochondria were deformed and in several mitochondria abnormal configuration of cristae was observed. The size of some organelles reached the 10-20 microns length with normal diameter. The elongated mitochondria were located either in the long axis of the fibers or in the transverse direction near to the Z discs. In rats receiving valproic acid intraperitoneally for 14 days in daily dose of 100 mg/kg, steatosis developed in the liver, and focal lipid droplets could also be found in the skeletal muscle. The mitochondria of the rat liver appeared as normal, but in muscle the structure and organization of the cristae were disoriented. These observations show that treatment with valproate causes a kind of drug-induced mitochondrial cytopathy with microvesicular lipid deposition in the muscle. The lipid deposits are likely a result of the inhibited mitochondrial fatty acid oxidation. The formation of the abnormal mitochondrial structure could serve as background for the visible consequences of affected lipid metabolism. PMID- 9413005 TI - Decrease of N-acetylaspartate after ACTH therapy in patients with infantile spasms. AB - Apparent brain atrophy has been frequently observed at CT and MRI after ACTH therapy in patients with infantile spasms. There are several hypotheses to explain ACTH-induced brain shrinkage: 1) a catabolic effect of ACTH on brain tissue, 2) a mineralocorticoid effect resulting in a loss of water and 3) an increase in cerebrospinal fluid (CSF) pressure compressing the brain. An average of 0.21 +/- 0.03 mg/kg of ACTH was administered to nine patients over a period of 14 to 17 days. Water content and concentrations of N-acetylaspartate (NAA), creatine and phosphocreatine (Cr + PCr), and choline (Cho) were measured before, immediately after, and several months after the ACTH therapy by using in-vivo 1H magnetic resonance spectroscopy (MRS). Only NAA concentration exhibited a significant change during the study (6.6 +/- 1.5 mmol/kg, 5.4 +/- 1.1, and 7.0 +/ 1.5, p = 0.017). There was no significant change in Cr + PCr, in Cho, or in water content. These data suggest catabolic effects of ACTH on brain tissue, such as cell loss, decrease in NAA synthesis in mitochondria, and leakage of NAA from cell membrane. PMID- 9413006 TI - Elevated nerve growth factor levels in cerebrospinal fluid associated with progressive cortical atrophy. AB - Using a two-site enzyme-linked immunosorbent assay, we measured nerve growth factor (NGF) levels in cerebrospinal fluid (CSF) from 40 patients with or without brain atrophy due to various neurologic disorders. White matter and cortical atrophies were assessed by frontal horn index (FHI) and subarachnoid space area/inner skull space area (SSA/ISSA) ratio, respectively. CT findings of 40 patients were classified into 4 grades: normal (< mean + 2SD), grade I (mean + 2SD < or = or < mean + 4SD), grade II (mean + 4SD < or = < mean + 8SD), and grade III (mean + 8SD < or =). NGF levels in CSF were significantly elevated in 3 of 6 patients with grade III cortical atrophy and normal in 2 with grade II atrophy, 4 with grade I atrophy and 28 without atrophy. Cortical atrophy was progressive in the 3 with NGF elevation in CSF. With respect to white matter atrophy, NGF elevation was observed in none of 3 with grade III white matter atrophy, two of 4 with grade II atrophy, none of 3 with grade I atrophy and one of 30 without atrophy. The symptoms of the two of three with NGF elevation were progressive at the time of obtaining the CSF samples, while those of other patients without NGF elevation were non-progressive. The present study suggests that NGF elevation in CSF may reflect extensive cortical degenerative change. PMID- 9413007 TI - Disorganized patterns: chronic-stage EEG abnormality of the late neonatal period following severely depressed EEG activities in early preterm infants. AB - EEG recordings were performed within 72 hours following the birth of 55 preterm infants with a gestational age of less than 29 weeks. Seventeen infants (31%) manifested moderately to severely depressed EEG activities during this period. Eight of the infants died within a few days after birth, and 9 survived. We recorded EEGs from the surviving infants serially throughout the neonatal period, and evaluated EEG findings in the recovery phase following depressed EEG activities. At 1 to 3 weeks after birth, EEGs revealed an abnormal morphology of background activities in 8 of 9 cases; this was also true of the EEGs of 6 cases at 4 weeks or more of postnatal age. The latter 6 cases demonstrated deep white matter injury on cranial ultrasonography, and all of them later developed cerebral palsy. Based on the foregoing, EEG findings of early preterm infants in the late neonatal period are considered a useful means to detect deep white matter injury and to allow neurological prognosis. PMID- 9413008 TI - Prognosis of hypoxic-ischaemic encephalopathy in full-term newborns--value of neonatal electroencephalography. AB - This study was performed to assess the predictive value of EEG in HIE following acute fetal distress. For 38 full-term neonates, EEG was recorded at least before 48 hours of life and between 2 and 7 days. Neurological outcome was evaluated at a minimum age of one year. Normal or slightly abnormal early EEGs (14 cases) were observed with normal outcome (13 cases) or minor sequelae (1 case). Extremely abnormal early EEGs were associated with death (5 cases), severe sequelae (4 cases) or normal outcome (1 case); among the "intermediate" group (14 cases), improvement of background activity before 7 days indicated a good prognosis (4/5), whereas identical or worsened activity was noted in all cases with a poor outcome. Of neonates 90% with very abnormal EEGs and 64.3% with "intermediate" EEG presented a significantly unfavourable outcome compared with neonates having normal recordings (p < 0.0001). The EEG before 48 hours has an excellent sensitivity rate (94.7%) but a less satisfactory specificity rate (68.4%). PMID- 9413009 TI - A neurophysiological study in children and adolescents with Crigler-Najjar syndrome type I. AB - We studied the neurophysiological features of five patients (age range: 4-20 years) suffering from Crigler-Najjar syndrome type I (CNsI) by means of multimodal (brainstem, somatosensory, motor) evoked potentials and periodic EEG polygraphic recordings (follow-up: 3 months-4.5 years). Two patients presented with neurological disturbances, consisting mainly of mental slowing, motor impairment and seizures. Both of them presented an abnormal EEG, characterized by slowing of background activity associated with paroxysmal discharges. Liver transplantation was performed in one of these two patients and was followed by improvement of both the neurological picture and EEG activity. In a third patient, clinically normal, after two years of follow-up, the EEG started to show paroxysmal activity during sleep or when evoked by intermittent photic stimulation. In these three patients, multimodal evoked potentials were unremarkable. The remaining two younger subjects did not show any clinical or EEG abnormality. Our findings suggest that, whereas in newborns and infants evoked potentials have been demonstrated as reliable techniques to monitor bilirubin neurotoxicity, in children and adolescents with CNsI, EEG seems to be more sensitive in evaluating patients for neurological damage and effectiveness of therapeutic strategies adopted. PMID- 9413010 TI - Infant botulism. The first culture-confirmed Danish case. AB - Infant botulism is caused by intestinal colonization by Clostridium botulinum, C. barati or C. butyricum. Infant botulism has only rarely been reported outside the USA. A 3-month-old boy developed constipation, lethargy, feeding difficulties and descending, severe, symmetric weakness. He was breastfed but had also been fed honey. Supportive care led to complete recovery. The serum was positive for C. botulinum toxin type A-F (mouse toxin neutralization assay). A strain of C. botulinum producing toxin type A and E was identified in the stool. C. botulinum was identified in a jar of honey of the same brand as the honey fed to the patient. PMID- 9413011 TI - Otoacoustic emissions as indicators of neurologically based hearing loss in childhood. AB - One of the most frequent causes of sensorineural hearing loss in childhood is damage to outer hair cells of the cochlea. The presence of otoacoustic emissions, generated by outer hair cells, provides evidence for normal hearing. This finding, however, may give rise to false reassurance, because even severe hearing loss, localized behind the cochlea, can be associated with normal otoacoustic emissions. The coexistence of otoacoustic emissions and hearing loss calls for the prompt exclusion of neurological disease. PMID- 9413012 TI - Thymectomy in children with generalized myasthenia gravis. PMID- 9413013 TI - Predominant basolateral proteolytic processing of prosomatostatin into somatostatin-28 in polarized LLC-PK1 cells. AB - Polarized epithelial cells secrete specific proteins through their apical or basolateral membrane. In the present study, we have expressed the human prosomatostatin cDNA in the pig kidney epithelial cell line (LLC-PK1) and monitored the processing and release of the somatostatin-related peptides. Analysis by high-performance liquid chromatography and radioimmunoassay of the somatostatin-related peptides synthesized by the transfected cells showed that the LLC-PK1 cells released prosomatostatin and somatostatin-28 (S-28) in the culture medium. Furthermore, when the cells were polarized, we observed release of prosomatostatin from both membrane domains (apical and basolateral), while liberation of S-28 was mostly from the basolateral side. This observation suggests that, in these cells, the proprotein convertase(s) responsible for prosomatostatin processing is(are) associated with the basolateral secretory pathway. PMID- 9413014 TI - Central and peripheral antiulcer and antisecretory effects of Ala5NKA(4-10), a tachykinin NK2 receptor agonist, in rats. AB - Analogs of NKA(4-10) which are selective tachykinin NK2 receptor agonists have been tested as anti-secretory and anti-ulcer agents in 2-h pylorus ligation in rats. Peripheral (500 micrograms/kg s.c.) or central (5 micrograms/rat i.c.v.) administration of Ala5NKA(4-10), but not NKA(4-10) or Ala5[beta Ala8]NKA(4-10), inhibited gastric ulcer and secretion. The same effective doses of Ala5NKA(4-10) did not influence gastric emptying. The anti-secretory and anti-ulcer effects of Ala5NKA(4-10) were antagonized by pretreatment with the tachykinin NK2 receptor antagonist MEN 10,627 at a dose (250 micrograms/kg s.c.) which did not affect gastric secretion and ulcers. These findings provide the first evidence that activation of central and peripheral tachykinin NK2 receptors affords protection against gastric ulcers induced by 2 h pylorus ligation in rats, by reducing gastric acid secretion. PMID- 9413015 TI - Relationship between binding affinity and functional activity of nociceptin/orphanin FQ. AB - The relationship between binding affinity and functional activity of nociceptin/orphanin FQ binding was studied in brain membranes, membranes of Chinese hamster ovary cells transfected with ORL1, and intact CHO-ORL1 cells. Binding affinities were compared with potency for the stimulation of [35S]GTP gamma S binding in cell membranes, and inhibition of forskolin-stimulated cAMP accumulation in intact cells. Binding was conducted with [3H]14-Tyr-nociceptin, and in brain or cell membranes the affinity was found to be 50-100 pM. The binding of [3H]14-Tyr-nociceptin was found to be regulated by Na+ and GTP, as expected for an opioid-like receptor. In intact cells, saturation produced a curvilinear Scatchard Plot. Non-linear analysis indicated two states of the receptor, with the vast majority of binding being to a low affinity state of approximately 8 nM. This low affinity component is consistent with the lower potency derived from the inhibition of cAMP accumulation, stimulation of [35S]GTP gamma S binding, and other functional assays. PMID- 9413016 TI - Comparison between vasoactive intestinal polypeptide and pituitary adenylate cyclase activating polypeptide levels in neuroblastoma tumour tissues. AB - Vasoactive intestinal polypeptide (VIP) is reported to exert an autocrine control on neuroblastoma cell tumours: VIP is produced by the tumour and stimulates cell differentiation. This study tested the hypothesis that the parent peptide; the pituitary adenylate cyclase activating polypeptide (PACAP) may have a similar role. It was found that PACAP mRNA and PACAP were expressed in 12/12 tumours; it was also observed that PACAP receptor mRNA and functional PACAP receptors were expressed in 12/12 and 5/9 tumours, respectively. VIP mRNA and VIP were detected in 9/12 tumours. VIP receptor mRNA was expressed in 5/12 tumours and functional VIP receptors were never demonstrated. The tumours having the highest VIP levels also had the highest PACAP contents and were associated with a watery diarrhoea syndrome due to activation of intestinal VIP receptors. As PACAP recognizes the PACAP receptors and the VIP receptors with the same high affinity it may contribute to the syndrome and is a likely candidate for an autocrine control of neuroblastoma cell growth and differentiation. PMID- 9413017 TI - Elevated cerebrospinal fluid level of substance P and decreased undecapeptidase activity at term pregnancy. AB - This paper reports a study of substance P and its converting enzyme substance P endopeptidase (SPE) in cerebrospinal fluid (CSF) collected from women at term pregnancy. A method was developed to assess the CSF levels of substance P itself with minimum contribution from prestages or fragments of the undecapetide. The measured activity was compared with that detected in CSF from control, non pregnant, non-puerperal women. The result indicates a significant increase in substance P-like immunoreactivity at term pregnancy (19.9 +/- 3.9 fmol/ml, n = 10) compared with that of control subjects (12.3 +/- 2.8 fmol/ml, n = 9; P < 0.001). This elevation was suggested to reflect an increased activity in spinal sensory neurons at this stage of pregnancy. The activity of SPE was assessed by measuring the product substance P1-7 by a radioimmunoassay specific for this fragment. It was found that the enzyme activity was significantly decreased in CSF from pregnant women (11.2 +/- 0.71 pmol/h.ml), compared with control samples (18.4 +/- 0.73 pmol/h.ml; P < 0.05). Moreover, a significant negative correlation was found to exist between the level of substance P and the activity of SPE (P < 0.05, r2 = 0.65), suggesting that the enzyme may be involved in a mechanism regulating the level of substance P concentration. PMID- 9413018 TI - Effect of kappa opioid agonist RU 51599 on osmotic and non-osmotic stimulated arginine vasopressin release and gene regulation in small cell lung carcinoma cells. AB - Arginine vasopressin (AVP) is synthesized in the hypothalamus, stored in the posterior pituitary, and osmotic and non-osmotic stimuli release AVP into the circulation for antidiuretic and vascular actions on target tissue. The kappa opioid agonist, RU 51599, exhibits a potent diuretic activity in both experimental animals and humans. This diuretic activity is characterized by a water diuresis without an associated increase in electrolyte excretion. Studies with cultured rat hypothalamo-neurohypophysial system explant showed that AVP mRNA level changed in parallel to the RU 51599-induced changes in AVP secretory rate. There are, however, no hypothalamic neuronal cell lines to study AVP gene regulation system, and it is not known whether RU 51599, regulates AVP secretion and biosynthesis under osmotic and non-osmotic stimulatory conditions of AVP release. The effect of RU 51599 on AVP release, AVP mRNA, and AVP gene promoter activity in osmotic and non-osmotic conditions was therefore studied using cultured small cell lung carcinoma (SCLC) cell lines. RU 51599 significantly inhibited AVP release by osmotic stimulation (330 mOsm) and non-osmotic stimulators, angiotensin II (AII) and endothelin 3 (ET3). However, RU 51599 did not show any effect on the AVP mRNA and AVP gene promoter activity stimulated by high osmolality and ET3. These results indicate, therefore, that RU 51599 suppresses AVP secretion by inhibition at the step of AVP release during osmotic and non-osmotic stimulation but does not affect the AVP gene transcription level in the SCLC cells. PMID- 9413019 TI - Vasopressin and oxytocin gene expression in the porcine forebrain under basal conditions and following acute stress. AB - This study, the first using the pig, examined expression of mRNAs for vasopressin (VP), oxytocin (OT), preproenkephalin (PENK) and pro-opiomelanocortin (POMC) in the forebrain, and of POMC and prolactin in the pituitary. High basal expression of VP and OT mRNAs was present in the paraventricular (PVN) and supraoptic (SON) nuclei. In the PVN, VP was found in magnocellular regions whereas OT was also seen in the parvocellular portion; the distribution of VP and OT mRNAs in the SON was as reported in other species. The suprachiasmatic nucleus contained VP mRNA but only OT message was present in the dorsomedial SON, a structure peculiar to swine. Gene expression for PENK occurred in the caudate putamen (CPu), for POMC in the mediobasal hypothalamus (MBH) and for prolactin and POMC in the hypophysis. Following restraint, VP message increased in the magnocellular PVN, as did PENK in the CPu and POMC in the MBH. PMID- 9413021 TI - Substance P is not involved in vascular nociception in humans. AB - Blood vessels are similar to skin invested with substance P-containing nerves and the binding sites for substance P, which has been shown to be involved in nociception from skin. No attempts have been made so far to see whether substance P is also involved in vascular nociception. We therefore tested substance P for its algesic properties in human veins because their innervation exclusively subserves nociception. Substance P never evoked pain from veins at concentrations of 10(-8)-10(-4) M but did so in a concentration-related fashion from skin. Therefore, substance P is not involved in the generation of acute vascular pain. PMID- 9413020 TI - Chronic gestational cocaine treatment decreases oxytocin levels in the medial preoptic area, ventral tegmental area and hippocampus in Sprague-Dawley rats. AB - We examined the effects of gestational cocaine treatment on oxytocin levels in the whole hippocampus (HIP), ventral tegmental area (VTA), medial preoptic area (MPOA) and amygdala (AMY) in rat dams on postpartum days (PPDs) 1 and 2. Cocaine treatment significantly reduced oxytocin levels in the MPOA within 12-16 h of delivery (PPD 1), but had no significant effect on the other brain areas. Oxytocin was significantly reduced in the HIP and VTA but not in the AMY or MPOA on PPD 2. These data provide the first evidence for the reduction of oxytocin levels in the VTA, HIP and MPOA as a result of gestational cocaine treatment. PMID- 9413022 TI - Biochemical and pharmacological activities of SR 144190, a new potent non-peptide tachykinin NK2 receptor antagonist. AB - (R)-3-(1-[2-(4-benzoyl-2-(3,4-difluorophenyl)-morpholin-2-yl)- ethyl]-4 phenylpiperidin-4-yl)-1-dimethylurea (SR 144190) is a new non-peptide antagonist of tachykinin NK2 receptors. SR 144190 potently and selectively inhibited neurokinin A binding to NK2 receptors from various species, including humans. In in vitro functional assays, it was a potent, selective and competitive antagonist of NK2 receptors with apparent affinities (pA2 values) between 9.08 and 10.10. In vivo, SR 144190 blocked [Nle10]neurokinin A-(4-10)-induced bronchoconstriction in guinea pigs (ID50 = 21 micrograms kg-1 i.v. and 250 micrograms kg-1 i.d.) and [beta Ala8]neurokinin A-(4-10)-induced urinary bladder contraction in rats (ID50 = 11 micrograms kg-1 i.v. and 190 micrograms kg-1 i.d.). It prevented citric acid induced cough and airway hyperresponsiveness to acetylcholine in guinea pigs (1 mg kg-1 i.p.) as well as castor oil-induced diarrhoea in rats (0.01-10 micrograms kg-1 s.c. or p.o). Finally, it blocked the turning behaviour induced by intrastriatal injections of [Nle10]neurokinin A-(4-10) in mice (ID50 = 3 micrograms kg-1 i.v. and 16 micrograms kg-1 p.o.). PMID- 9413023 TI - Failure of gammahydroxy butyric acid to stimulate growth hormone secretion in cocaine addicts. AB - This study discusses the effect of gammahydroxy butyric acid (GHB) on growth hormone (GH) secretion changes in cocaine addicts. Ten male cocaine users and 10 normal controls were tested with a single oral administration of GHB at a dose of 25 mg/kg body weight. Cocaine addicts were tested before and after 30 days of abstinence. All subjects underwent a control with a placebo. Basal GH levels were similar in normal controls and cocaine users and remained unmodified during the control test. In the normal control subjects, plasma GH levels rose significantly after the administration of GHB; in contrast, plasma GH concentrations failed to increase after GHB treatment in cocaine addicts. These data show that a chronic abuse of cocaine induces alterations of the GABAergic system which were unmasked by the absent GH response to GHB. PMID- 9413024 TI - Role of adrenal and gonadal steroids in the response of GnRH gene expression to the endogenous benzodiazepine receptor ligand octadecaneuropeptide in the male rat brain. AB - We have recently demonstrated that the inhibitory influence of the endogenous benzodiazepine receptor ligand octadecaneuropeptide (ODN) on gonadotropin releasing hormone (GnRH) gene expression could be prevented by specific inhibitors of 3 beta-hydroxysteroid dehydrogenase and 5 alpha-reductase in adrenalectomized and castrated male rats, then suggesting an involvement of neurosteroids in the action of ODN on GnRH neurons. In order to study in detail the role of circulating steroids in the effect of ODN, we have evaluated the influence of adrenalectomy, castration and the combination of adrenalectomy and castration as well as the effect of dexamethasone administration in the response of GnRH gene expression to ODN in the male rat. The intracerebroventricular injection of ODN (4 h before sacrifice) produced a 36% decrease in the hybridization signal. Adrenalectomy induced a 21% decrease in GnRH mRNA levels. In the adrenalectomized rats, the injection of ODN increased by 11% the amounts of mRNA. As previously reported by our group, castration was found to enhance GnRH mRNA (15% over control values). In castrated animals, ODN produced an inhibitory effect in the hybridization signal which was of the same amplitude as that observed in sham-operated animals. Finally, the combination of adrenalectomy and castration resulted in a small but significant decrease in the hybridization signal, the values being intermediary between those observed after adrenalectomy and those obtained after castration. In these animals, ODN induced a 38% decrease in the amounts of GnRH mRNA. In animals that had been castrated and adrenalectomized, dexamethasone treatment during 4 days produced a 19% increase in hybridization signal. In these dexamethasone-treated animals, ODN produced the usual decrease (33%) in GnRH mRNA. These results demonstrate that gonadal hormones do not play a major role in the activation of the GABAA receptor complex by ODN. On the other hand, it clearly appears that glucocorticoids exert a tonic stimulatory influence on GnRH neuronal activity and are involved in the inhibitory effect of ODN. The mechanism of action of glucocorticoids, which seems complex since the influence of adrenalectomy on the ODN action can be prevented by orchidectomy, remains to be fully elucidated. PMID- 9413025 TI - Dose-dependent inhibitory effects of angiotensin II on visual responses of the rat superior colliculus: AT1 and AT2 receptor contributions. AB - Angiotensin II (Ang II) has traditionally been regarded as a peripherally circulating and acting hormone involved in fluid homeostasis and blood pressure regulation. With the rather recent localization of Ang II receptors within the mammalian brain, renewed interest has emerged in the hope of elucidating the central impact and function of this hormone. One region that has been clearly demonstrated to express Ang II receptors is the superior colliculus (SC). This mesencephalic structure plays an important role in sensory visuomotor integration. Receptors for Ang II (of both the AT1 and AT2 subtypes) have been localized within the superficial layers of this structure, i.e. the areas that are visually responsive. In the hopes of characterizing the role of Ang II in the SC, we have attempted to physiologically activate these receptors in vivo and observe the effects of Ang II on visually evoked responses. In the attempt to identify the receptor subtype(s) responsible in mediating these effects, Ang II was injected concomitantly with selective receptor ligands. Experiments were performed on adult rats prepared in classical fashion for electrophysiological studies. Through microinjection of Ang II, and the simultaneous recording of visually evoked potentials to flash stimulation, we have observed that this peptide yields a strong suppressive effect on visual neuronal activity. By injecting Ang II at various concentrations (10(-3)-10(-10) M), we have further observed that the effects of this peptide express a dose-related dependency. Injection of Ang II in progressively more ventral layers yielded less pronounced effects, demonstrating physiologically the discrete localization of these receptors in the stratum griseum superficiale. Coinjection of Ang II with Losartan yielded a near complete blockade of Ang II suppressive effects, suggesting that AT1 receptors play a prominent role in mediating these responses. However, coinjection of Ang II with PD 123,319 yielded a slight, yet significant partial blockade. Coinjection of Ang II with both the AT1 and AT2 receptor antagonists yielded a complete blockade of the Ang II effect. Finally, some of the results suggest that the AT2 receptor ligand CGP 42,112 may possess agonist properties. Taken together, these findings suggest that the AT1 receptor is predominantly involved in mediating Ang II responses in the SC and there also appears to be some indication of AT2 receptor involvement. However, the underlying mechanisms (such as receptor interactions), the exact specificity of the ligands used, and the possibility of other receptor subtype implication have yet to be explored fully. PMID- 9413026 TI - Synthesis and binding characteristics of the highly selective radiolabelled deltorphin analogues containing 2-aminotetralin-2-carboxylic acid in position 3. AB - Following the description of [3H]Ile5,6deltorphin II, when it was reported that changes in hydrophobicity at positions 5 and 6 give rise to analogues with increased delta-receptor affinity and selectivity, new conformationally restricted deltorphin analogues were designed. A synthetic amino acid, 2 aminotetralin-2-carboxylic acid (Atc), was introduced at position 3 instead of Phe in Ile5,6deltorphin I and II, and the resultant compounds were prepared in tritiated form. Opioid binding sites specific for [3H]S-Atc3,Ile5,6deltorphin I and [3H]R-Atc3,Ile5,6deltorphin II were characterized in rat brain membranes. Their binding was saturable, stereoselective and inhibited by delta-selective ligands with high potency. They labelled single class of opioid sites at 35 degrees C with high affinity (Kd approximately 0.3 nM), Bmax values of 130 fmol/mg protein, and very low non-specific binding was observed. Both tritiated deltorphin analogues showed delta-receptor specificity in rat brain, therefore they could represent excellent new radioligands for investigating the complexity of the opioid receptor systems. PMID- 9413027 TI - Vasopressin fragment, AVP-(4-8), improves long-term and short-term memory in the hole board search task. AB - The hole board search task (HBST) measures long-term and short-term memory, operationally defined as reference memory and working memory. The HBST is an open field spatial learning test. Previously, we have shown that desglycinamide(Arg8) vasopressin (DGAVP) modulated reference memory, working memory, spatial sequence memory, and learning in the HBST in a dose-dependent manner (Vawter MP, Van Ree JM. Effects of des-glycinamide-sup-9-(arginine-sup-8) vasopressin upon spatial memory in the hole-board search task. Psychobiology 1995; 23: 45-51). To examine the potential active site of the DGAVP molecule, the fragment of the vasopressin amino acid sequence, [pGlu4,Cyt6]AVP-(4-8) (AVP-(4-8)), was administered 1 h prior to training in the HBST. Three groups received either 0, 0.3 microgram, or 1 microgram AVP-(4-8). A repeated measures MANOVA showed the AVP-(4-8) pretreatment factor to be significant (P = 0.048) on the reference memory measure, but not the working memory or learning measures. Interactions between peptide x sessions for reference memory (P = 0.015), working memory (P = 0.003) and learning (P = 0.010) indicated differences in improvement over sessions between placebo- and peptide-treated groups. Post hoc comparisons revealed that the AVP-(4-8) fragment in a dose of 0.3 microgram increased reference memory on the fourth, fifth and sixth acquisition sessions compared with placebo or 1 microgram AVP-(4-8) pretreated groups. Working memory and errors were significantly lowered by 0.3 microgram AVP-(4-8) on the first acquisition session when compared with placebo pretreatment. Thus, AVP-(4-8) improves long-term and short-term memory scores in the HBST, similar to previous results with DGAVP. However, AVP-(4-8) appears twice as potent than DGAVP in improving long-term memory scores in the HBST. The data suggest that the memory modulating property of DGAVP is contained within the amino acid sequence of the AVP-(4-8) peptide. PMID- 9413028 TI - Tachykinin receptors on motoneurons in the spinal cords of neonatal rats, gerbils and hamsters. AB - As a step to clarify the profiles of tachykinin receptors in the mammalian central nervous system, we examined the effects of various tachykinin receptor agonists and antagonists on motoneurons in isolated spinal cord preparations from rats, gerbils and hamsters. After treatment with tetrodotoxin, potential changes were recorded extracellularly from lumbar ventral roots at 27 degrees C. Bath application of tachykinin NK1, NK3 receptor agonists produced depolarizing responses of ventral roots. In contrast, selective NK2 agonists exerted no or only marginal depolarizing action. Neurokinin A (NKA), however, exerted a distinct depolarizing action on motoneurons. Tachykinin NK1 receptor antagonists antagonized the actions of SPOMe and NKA in a competitive manner. The present results suggest that tachykinin NK1 and NK3 receptors are present on spinal motoneurons of newborn rats, gerbils and hamsters, and that NKA acts on the NK1 receptors. PMID- 9413029 TI - The antiproliferative effects of calcitonin gene-related peptide in different passages of cultured vascular smooth muscle cells. AB - Previously our laboratory showed that calcitonin gene-related peptide (CGRP) is released from perivascular nerves exposed to endotoxin or inflammatory mediators bradykinin and prostaglandins. CGRP contributes significantly to the vasodilation of inflammation and septic shock. Another potential action of CGRP is inhibition of vascular smooth muscle cell (VSMC) growth, which could serve to counterbalance the stimulatory effects of IL-1 and TNF on VSMC proliferation. VSMCs from rabbit and rat aorta (in the second and fifth passages) were plated at 100,000 cells/well in 24-well trays in 10% fetal bovine serum (FBS) for 24 h, incubated for an additional 24 h without FBS, and then exposed to 2.5% FBS for 24 h in the presence or absence of CGRP. 3H-thymidine incorporation was used to measure DNA synthesis and proliferation. CGRP caused significant inhibition of 3H-thymidine incorporation, which correlated with elevations of cAMP in both rat and rabbit aortic VSMCs. Interestingly, the responses of both the elevation of cAMP and the inhibition of DNA synthesis became larger in VSMCs with an increasing number of passages. The data suggest that the CGRP, released during vascular inflammation, may serve to inhibit the proliferation of VSMCs, thus limiting the growth of atheromatous lesions. PMID- 9413030 TI - Effects of chronic U-50,488H treatment on the isolated right atrium of the rat. AB - The aim of the present investigation was to determine if chronic activation of kappa-opioid receptor induces development of tolerance and dependence to kappa opioid agonists on the isolated right atrium of the rat. Tolerance to the kappa agonist was induced by chronic administration of U-50,488H, a selective kappa agonist (15 mg/kg i.p. twice a day for 4 days). The rats were killed on day 5. Tolerance to U-50,488H was observed after its chronic administration and was revealed as a rightward shift of the concentration-response curve; it was accompanied by a decrease in the maximum response and in the slope. Withdrawal to the kappa-agonist was induced by administration of Mr-2266 (preferentially kappa antagonist) or nor-binaltorphimine (nor-bni; selective kappa-antagonist) to the organ bath. The administration of the kappa-antagonists Mr-2266 or nor-bni to preparations from tolerant rats in the organ bath induced an increase in auricular contraction frequency. In contrast, the administration of the kappa antagonists to preparations from vehicle-treated rats induced a decrease in auricular contraction frequency. These findings demonstrate that the hearts of rats that had received chronic U-50,488H treatment develop tolerance to the cardiac effects of U-50,488H and exhibit excitatory reactions to kappa antagonist's precipitated withdrawal. PMID- 9413031 TI - beta-endorphin inhibits and facilitates lordosis behaviour in rats depending on ventricular site of administration. AB - beta-endorphin was administered intracerebroventricularly into the lateral and third ventricles of ovariectomized, oestrogen- and progesterone-primed rats, and its effect on lordosis and ear-wiggling was assessed. A dose of 2 micrograms beta endorphin facilitated lordosis when infused into the lateral ventricle, but inhibited lordosis when infused into the third ventricle. The effects were the same whether measured at 30, 60 or 90 min following infusion. beta-endorphin had no significant effect on ear-wiggling frequency when administered in either ventricle. The differential effects of beta-endorphin depending on site of administration may reflect the activation of distinct opioid receptor subtypes within the brain. PMID- 9413032 TI - Genetic changes in prostate cancer. AB - Recent advances in molecular biology have allowed us to understand that it is the accumulation of genetic alterations which leads to each step of tumorigenesis. What the specific alterations may be, however, often varies with each neoplasm. Prostate cancer is somewhat unique in its presentation to the pathologist of a bewildering array of histologies difficult to assign to diagnostic categories and contributing to misinterpretations of underlying molecular events. As with any malignancy, it is of utmost importance to thoroughly analyze and record the genetic aberrations found in prostate cancer with the objective of correlation to the pathology and natural history of the disease. Multiple oncogenes and tumor suppressor genes have been investigated in both clinical and latent cancer using conventional mutational analyses. To probe deeper into these genes and to uncover novel molecular events, genomic tumor DNA were examined using restriction landmark genomic scanning (RLGS), a method which allows the identification and comparison of specific genetic alterations within large segments and multiple samples of DNA at a time. This article reviews what has been identified based on numerous molecular studies, focusing on the genetic alterations peculiar to human prostate cancer. PMID- 9413033 TI - Causality of parenchymal and vascular changes in rats with experimental thiamine deficiency encephalopathy. AB - The causality of vascular and parenchymal damage to the central nervous system (CNS) was examined in rats with thiamine deficiency. Male Sprague-Dawley rats were divided into two groups; one was given a thiamine-deficient diet (TDD) and injected intraperitoneally with 10 micrograms/100 g bodyweight pyrithiamine (PT) in order to analyze morphometrically the topographical and sequential relationship between vascular and parenchymal changes and vasodilatation, and the other was given a TDD and 50 micrograms/100 g bodyweight PT in order to determine hemorrhagic sites using serial sections. Histological examination showed that spongiotic change occurred selectively in the inferior colliculus (100%) from day 19, and thereafter in the thalamus (95%), mammillary body (50%) and nuclei olivaris and vestibularis of the pons (25%), with or without hemorrhage. Simultaneously, glycogen accumulation was also observed in these regions at a frequency similar to that of hemorrhage. Ultrastructurally, however, hydropic swelling of astrocytic and neuronal processes without glycogen accumulation was observed as early as day 9 in the inferior colliculus, at which time an increase of glial fibrillary acidic protein-positive processes was also recognized. The superior colliculus was completely spared. From day 22 vasodilatation of the inferior colliculus occurred, concomitantly with bodyweight loss and neurological symptoms. Twenty-two examined hemorrhages, which occurred in the thalamus and inferior colliculus, were distributed along the arterioles or capillaries on the arterial side. In conclusion, the morphological CNS changes caused by thiamine deficiency with administration of low-dose PT in rats begin as hydropic swelling of neuronal and astrocytic processes, followed by hemorrhage and, thereafter, by vasodilation. The predilection for hemorrhage on the arterial side without parenchymal changes suggests that petechial hemorrhage is not simply secondary to parenchymal changes, but is due to hemodynamic change resulting from thiamine deficiency-induced vascular dysfunction. PMID- 9413034 TI - Expression of Bcl-2 in human breast cancer: correlation between hormone receptor status, p53 protein accumulation and DNA strand breaks associated with apoptosis. AB - The expression of Bcl-2, a suppressor of apoptotic cell death, was investigated in 52 invasive carcinomas of the breast using reverse transcription-polymerase chain reaction and immunohistochemical methods. After consideration of both sets of results, 42 tumors (80.8%) were confirmed to be positive (Bcl-2(+)) and 10 (19.2%) were judged negative (Bcl-2(-)) for Bcl-2 expression. Related factors (p53 protein accumulation, hormone receptor status and apoptotic cell index) were also examined using immunohistochemical and in situ end-labeling methods to elucidate their correlations with Bcl-2 expression. Bcl-2 expression correlated significantly with the hormone receptor status, whereas it showed significant inverse correlations with p53 accumulation and the apoptotic index. It was concluded that estrogen and mutant p53 are related to the regulation of Bcl-2 expression and that the ability to prevent tumor cell death due to Bcl-2 can be developed by breast cancers. PMID- 9413035 TI - Bile canaliculi-like lumina in fibrolamellar carcinoma of the liver: a light- and electron-microscopic study and three-dimensional examination of serial sections. AB - Fibrolamellar carcinoma (FLC) is a variant of hepatocellular carcinoma characterized by distinct pathological features. The presence of intracellular lumina resembling bile canaliculi was previously reported in tumor cells of FLC on electron microscopy. Using light microscopy, we describe the presence of intracellular lumina in FLC, which was resected from a 15-year-old Japanese girl, as round structures lined with a brush-like border. These lumina occasionally contained bile. Light microscopic examination of 1 micron thick serial sections of Epon-embedded tissue samples showed that the lumina were located in the intracellular space without any connection to the intercellular space. However, we also detected a small number of lumina that were lined by microvilli, which were present between adjacent tumor cells. Results suggest that the presence of the intracellular lumina in tumor cells probably represents a common histopathologic feature of FLC. PMID- 9413036 TI - Morphogenesis and development of superficial spreading tumor of the colon and rectum. AB - The aim of this study was to clarify the morphogenesis and mechanism of a wide intramucosal extension of a superficial spreading (epithelial) tumor (SST; defined as an epithelial tumor with wide intramucosal spreading involving a diameter of 30 mm or more) in the colon and rectum. For this purpose favorable sites, histological components, and histological growth patterns were compared between 95 cases of SST (16 adenomas and 79 carcinomas) and 2356 non-SST cases, which served as controls. The frequency of SST was significantly higher in the cecum and rectum, and lower in the sigmoid colon when compared to the locations of the control. Among the SST cases, 82.3% of superficial spreading carcinoma (SSC) had an adenomatous component and 96.2% had a cytologically low-grade carcinoma (CAL). In intramucosal SSC, the adenomatous and/or CAL component was predominant, and the proportion of high-grade carcinoma (CAH) was significantly smaller in intramucosal SSC in comparison with the control group. In the mucosal spreading area of SST, 78.9% were tubulovillous in histological type and 86.3% showed a replacing growth pattern. These results indicate that an SST initially develops as an adenoma in at least 85.3% of cases and CAL in 14.7% cases at most; spreads superficially in the mucosa by a replacing growth mechanism that forms a tubulovillous and villous structure; and is affected by intestinal peristalsis less than non-SST. PMID- 9413037 TI - Adenoid basal carcinoma of the cervix uteri: a case report. AB - We report a rare case of adenoid basal carcinoma of the uterine cervix, unexpectedly found in a uterus resected for the treatment of cervical intraepithelial neoplasia (CIN) 3. The patient was a 47-year-old Japanese female. She received a total abdominal hysterectomy under a diagnosis of CIN 3 of the cervix. Grossly, there were no significant findings in the surgical specimens. Microscopically, in seven of the 12 blocks of the cervix examined, scattered small nests of uniform small cells, which extended 4 mm below the epithelial surface, with dark nuclei and scant cytoplasm were observed. Peripheral palisading as well as the formation of gland-like or acinar structures were noted. The latter were positive for mucicarmine. Stromal reaction was not obvious. There were also foci of squamous differentiation in some portions of the small nests. Occasional mitoses as well as large atypical cells were also seen in this area. Immunohistochemically, the foci of squamous differentiation were positive for carcinoembryonic antigen. The epithelial surface in other portions showed CIN 3 with crypt extension. Distinction between adenoid basal carcinoma of the cervix and other diseases, such as adenoid cystic carcinoma and squamous cell carcinoma with basaloid features, is important for clinical management because the clinical behavior of adenoid basal carcinoma is less malignant. PMID- 9413038 TI - Primary renal angiosarcoma: a case report and review of the literature. AB - Primary renal angiosarcoma is very rare. To our knowledge, only 15 cases have been reported to date. A 77-year-old Japanese man with a unilateral kidney presented with massive hematuria followed by renal failure. A renal tumor was suspected and a left nephrectomy was performed. The histopathological diagnosis was angiosarcoma of the kidney. A hemorrhagic tumor measuring 10 x 5 cm and clotted blood was found in the medullary area. The atypical tumor cells had a sinusoidal and solid appearance, and showed immunohistochemically positive reactions for some of the endothelial markers. The patient died about 21 months after the nephrectomy and the autopsy revealed massive metastases to the liver and retroperitoneum. One of the differential diagnoses of the case was angiomyolipoma, because the tumor cells were relatively bland in their histological appearance with entrapped fat cells in the pelvic area. Fifteen case reports with titles that included the term 'hemangiosarcoma/angiosarcoma', 'hemangioendothelioma/endothelioma' or 'vascular sarcoma' of the kidney were reviewed and compared to the present case. PMID- 9413039 TI - Pleomorphic hyalinizing angiectatic tumor of soft parts. AB - A case of recently described pleomorphic hyalinizing angiectatic tumor (PHAT) of soft parts is reported. The subcutaneous solid tumor arising in the axilla of a 58-year-old man was histologically characterized by sheets of mitotically inactive oval and pleomorphic cells, intranuclear cytoplasmic inclusions, and clusters of ectatic vessels with perivascular hyalinization. Mono- and multinucleated giant cells were also present. A hemangiopericytoma-like pattern of vascularity, pseudovascular spaces, stromal collagen with degenerative changes, and immunoreactivity for CD34 were observed. Since these features were very similar to those of solitary fibrous tumors of various sites and newly categorized giant cell angiofibroma, it is considered that PHAT, solitary fibrous tumor, and giant cell angiofibroma may be in the same family of tumor. The tumor was diploid with a low S-phase fraction. The patient was well with no evidence of disease for 23 months. PMID- 9413040 TI - Numerous eosinophilic globules (skeinoid fibers) in a duodenal stromal tumor: an exceptional case showing smooth muscle differentiation. AB - A unique case of duodenal stromal tumor in a 51-year-old man is reported. The tumor histologically showed spindle cell proliferation and numerous eosinophilic globules. Most globules were composed of tangled 45 nm thick fibrils, which were ultrastructurally identical to 'skeinoid fibers'. The presence of glycogen granules in the tumor cells and the immunoreactivity for alpha-smooth muscle actin suggested smooth muscle differentiation. Focal ultrastructural findings also supported the smooth muscle nature of this tumor. There were no immunohistochemical and ultrastructural features indicating neural differentiation. In previous studies, the presence of such 'skeinoid fibers' was suggested to be a histological marker for neural differentiation in gastrointestinal stromal tumor. However, the findings in the present case suggest that numerous 'skeinoid fibers' can be identified in duodenal stromal tumor with smooth muscle differentiation, although this condition may be rare. PMID- 9413041 TI - Jejunal stromal tumor with skeinoid fibers or myenteric plexoma: a case report. AB - Small intestinal stromal tumors with 'skeinoid fibers' are uncommon stromal tumors with an associated controversial histogenesis. Although their microscopic appearance is suggestive of a smooth muscle nature, they lack specific smooth muscle features, as evident by electron microscopy and immunohistochemistry. They also appear to lack features of neurogenic origin because they fall to react with neural/neuroendocrine markers such as S-100 protein, neuron-specific enolase and chromogranin. It is interesting, nonetheless, to note that the ultrastructural examination of these tumors may show structures reminiscent of neural differentiation, such as cytoplasmic projections, containing occasional membrane bound, dense-core, neurosecretory-type granules, which mimick the long cytoplasmic processes seen in tumors of neural origin. Moreover, the association of these tumors with Von Recklinghausen's neurofibromatosis, as well as the presence of 'skeinoid fibers' in proven neurogenic spindle cell neoplasms such as gastrointestinal autonomic nerve tumors and schwannomas, suggests that these tumors might also be neurogenic in origin and enhances the diagnostic value of 'skeinoid fibers' as a possible ultrastructural marker of neural differentiation. Thus, light microscopic evaluation is clearly insufficient to accurately diagnose these tumors and to determine their histogenesis, electron microscopic and immunohistochemical studies being necessary. In this article the histogenesis of small intestinal stromal tumors with 'skeinoid fibers', regarding a jejunal neoplasm in a 63-year-old patient, is reviewed. The light microscopic, immunohistochemical and ultrastructural features are described and compared with findings usually seen in all those stromal tumors which may raise a differential diagnosis, such as smooth muscle stromal tumors, gastrointestinal autonomic nerve tumors, schwannomas, paragangliomas and fibrosarcomas. PMID- 9413042 TI - Epstein-Barr virus-related Hodgkin's disease showing B cell lineage in an immunosuppressive patient seropositive for HTLV-I. AB - A case of Hodgkin's disease (HD), lymphocyte depression (LD) type in an immunosuppressive patient is described. The patient was a 48-year-old male and his parents were born in the Kyushu area, which is an endemic area for adult T cell lymphoma/leukemia (ATL). He was seropositive for ATL virus (ATLV, also referred to as HTLV-I) and showed a marked immunosuppressive condition. He developed LD-HD and Pneumocystis carinii pneumonia, and died due to respiratory failure. The immunohistochemical and in situ hybridization analyses revealed that the Reed-Sternberg-like cells in the lymph node biopsy sample were positive for Ber-H2 (CD30), Leu-M1 (CD15), L-26 (CD20), Bcl-2, p53 and EBER, the viral genome of Epstein-Barr virus (EBV). PMID- 9413043 TI - 'Ovarian-type' stroma of pancreatic mucinous cystic tumor expresses smooth muscle phenotype. AB - 'Ovarian-type' stroma of nine mucinous cystic tumors (MCT) of the pancreas, six cystadenomas and three cystadenocarcinomas, were studied immunohistochemically. Most of the spindle cells of the 'ovarian-type' stroma were positive for smooth muscle actin and desmin, but were negative for S-100 protein, cytochrome P450 19 and CD34. Similar stroma with smooth muscle differentiation has been described in hepatobiliary MCT. Embryologic similarity between the pancreas and liver suggests a closely related origin of MCT in both organs. PMID- 9413044 TI - Recommendations for the reporting of larynx specimens containing laryngeal neoplasms. Association of Directors of Anatomic and Surgical Pathology. AB - The Association of Directors of Anatomic and Surgical Pathology have developed recommendations for the surgical pathology reporting of common malignant tumors. The recommendations for carcinoma of the larynx are reported herein. PMID- 9413046 TI - Initial treatment of severe acute pancreatitis. PMID- 9413045 TI - Calcifying fibrous pseudotumor after trauma. PMID- 9413047 TI - Clinical application of the intra-aortic balloon pumping apparatus and its balloon in Japan. PMID- 9413048 TI - The role of motilin and cisapride in the enteric nervous system of the lower esophageal sphincter in humans. AB - To assess the pharmacophysiological significance of the enteric nervous system and the responses of the human lower esophageal sphincter (LES) to motilin and cisapride, the mechanical responses of esophageal tissues from six patients with esophageal cancer and seven patients with gastric cancer were investigated. Circular muscle reactions were recorded to evaluate the in vitro esophageal responses to electrical field stimulation (EFS), motilin, and cisapride, evoking the adrenergic and cholinergic nerves before and after treatment with various autonomic nerve blockers. The findings of this study revealed that: cholinergic nerves are mainly involved in the regulation of enteric nerves in the steady state, while non-adrenergic non-cholinergic (NANC) inhibitory nerves also exist; motilin may act both via nerves and also directly on the LES smooth muscle; and cisapride releases acetylcholine from the end of the postganglionic fiber of the cholinergic nerve in human LES thereby inducing contraction of the LES. These results suggest that cholinergic and NANC inhibitory nerves play an important role in human LES, and that motilin and cisapride is clinically useful for improving the impaired LES of patients with gastroesophageal reflux. PMID- 9413049 TI - The operative indications for proximal gastrectomy in patients with gastric cancer in the upper third of the stomach. AB - While proximal gastrectomy is often performed for early gastric cancer in Japan, it remains unclear whether or not proximal gastrectomy should be performed for advanced gastric cancer. This study was designed to determine the operative indications for proximal gastrectomy in patients with gastric cancer in the upper third of the stomach. A total of 1691 patients with gastric cancer were reviewed retrospectively from hospital records during the period from 1969 to 1994, and the clinicopathologic characteristics of 82 patients who underwent proximal gastrectomy were compared with those of 150 patients who underwent total gastrectomy. Lymph node metastasis along the lower part of the stomach was observed in gastric cancers which had invaded beyond the muscularis propria of the stomach, but not in those confined to the muscularis propria. Three patients with gastric cancer that had invaded beyond the muscularis propria and metastasized to nodes along the lower part of the stomach were cured by total gastrectomy. However, there was no difference in the postoperative survival rates of the patients treated with proximal gastrectomy and those treated with total gastrectomy, irrespective of tumor stage and depth of invasion. Thus, proximal gastrectomy should be performed for gastric cancer when the depth of invasion is confined to the muscularis propria of the stomach. PMID- 9413050 TI - Analysis of p53 gene deletions in colorectal cancers using fluorescence in situ hybridization. AB - To examine the relationship between the incidence of p53 gene deletion in each nucleus and the clinicopathological features in colorectal cancers, we performed a cytogenetic study using fluorescence in situ hybridization (FISH). FISH was performed on 5 adenomas and 38 colorectal cancers that had been resected surgically. The nucleus, in which the copy number of the p53 signal was lower than that of chromosome 17, was determined as a deletion of the p53 gene. The mean frequency of the deletion of p53 in adenomas and cancers were 7.8% +/- 3.0% and 57.0% +/- 19.0%, respectively. Numerical aberrations of chromosome 17 or a deletion of p53 were also detected in DNA diploidy. The mean frequency of the deletion of p53 in 32 cases with aneusomy of chromosome 17 (65.7% +/- 14.5%) was significantly higher than that in cases of disomy (51.1% +/- 19.3%, P < 0.05). Even though this frequency was high in the early stage, it was not associated with any specific histopathological features. This frequency was also higher in double primary cancers (70.4% +/- 16.7%) compared with single colorectal cancers (53.4% +/- 18.1%) (P < 0.05). Using FISH, our results demonstrated that the clonal deletion of the p53 locus is an early genetic event of colorectal cancers and that a high incidence of p53 deletion may influence the occurrence of double primary cancers. PMID- 9413051 TI - An assessment of the anatomical relationship between the pelvic plexus and the rectal wall to determine the indications for its preservation in surgery for rectal cancer. AB - Preservation of the pelvic plexus in surgery for rectal cancer could shorten the distance between the cancer and the lateral resection margin, whereby the curability of the operation may be reduced. To clarify the indications for preserving the pelvic plexus in such surgery, the relationship of the pelvic plexus to the rectum and rectal cancer was investigated anatomically in 12 autopsied specimens and 12 surgical specimens. The rectum and anus were dissected with all the pelvic organs from autopsied cadavers and transverse sections were prepared at 10-mm intervals after fixation. The location of the pelvic plexus was then measured on the tissue preparations, and compared to that of surgical specimens from rectal cancers with concurrent resection of the pelvic plexus. The pelvic plexus was located from 3.3 +/- 1.2 cm above to 2.3 +/- 1.9 cm below the peritoneal reflection in the autopsied specimens. The average distances between the muscularis propria and the pelvic plexus in the autopsied specimens and surgical specimens were 8.3 +/- 3.5 mm and 14.7 +/- 4.5 mm, respectively, showing a significant difference (P < 0.05). Pelvic plexuses were located about 10 mm from the outer margin of rectal muscularis propria. These findings indicate that concurrent resection of the pelvic plexus may be required to secure sufficient surgical clearance in pT3 rectal cancers, especially those invading deeply beyond the muscularis propria (a2). PMID- 9413052 TI - Does bile spillage during an operation present a risk for peritoneal metastasis in bile duct carcinoma? AB - Whether bile spillage during operation presents a risk for peritoneal metastasis in the treatment of bile duct carcinoma was studied in 15 patients (12 with bile duct cancer, 3 with cancer of the papilla of Vater) who had all undergone a pancreatoduodenectomy. Preoperative bile was sampled through a percutaneous transhepatic biliary drainage catheter. Nine patients with bile duct cancer and one with cancer of the papilla of Vater showed positive bile cytology. The operative bile was obtained at the hepatic duct stump after a resection of the tumor-bearing bile duct. The operative bile in 10 patients with positive preoperative bile was found to be positive, while that in the five patients with negative preoperative bile was negative. Thus, the specificity of operative bile was identified as 100%. Moreover, in five patients with preoperative positive bile, saline irrigation of intrahepatic bile duct after a full recovery of hepatic bile revealed cancer cells to remain in the intrahepatic biliary trees. The viability of preoperative bile was 61%-97% with 1 x 10(4)-2.4 x 10(5) tumor cells, whereas there was a 41%-97% viability with 7.6 x 10(4)-10.4 x 10(5) tumor cells in the operative or irrigated bile. Accordingly, the patients with preoperative positive bile are thus suggested to be at high risk of inducing peritoneal metastasis due to the inadvertent spillage of hepatic bile at the time of resection of a bile duct tumor. PMID- 9413053 TI - An evaluation of splenopancreatic disconnection as a modification of the distal splenorenal shunt, studied in nonalcoholic patients by sequential angiography. AB - To evaluate the validity and complications of modifying the distal splenorenal shunt (DSRS) by performing splenopancreatic disconnection (SPD), hemodynamic changes in the portal system were assessed by visceral angiography in 93 patients with nonalcoholic portal hypertension during early postoperative follow-up after DSRS. There were 40 patients who underwent DSRS alone and 53 who underwent DSRS plus SPD. Early follow-up angiography showed that portal vein perfusion was well maintained, and that the diameter of the portal vein had decreased significantly by the same degree in both groups. Hepatofugal collaterals for the shunt had developed to a greater extent in the DSRS group, while they were almost completely absent in the DSRS with SPD group. Nevertheless, partial portal vein thrombosis was not detected in the DSRS group, although it was seen in seven (13.2%) of the patients who underwent DSRS plus SPD, in whom the left proximal splenic vein was not visible. The proximal splenic vein was seen in significantly less of the DSRS with SPD patients (47.2%) than the DSRS group patients (85%). In conclusion, SPD more effectively prevented the early postoperative development of collateral pathways for the shunt compared with standard DSRS; however, the possible stagnation of blood flow in the left proximal splenic vein may predispose to a risk of partial portal vein thrombosis developing during the early postoperative period after DSRS with SPD. PMID- 9413054 TI - A new preperitoneal repair for inguinal hernia using a transpositioned external oblique aponeurotic flap. AB - This study consists of a preliminary report of 94 cases with various types of inguinal hernias. All cases were repaired by a new technique, in which the herniotomy is performed via a preperitoneal approach and the repair is achieved by using a bipedicled flap from the external oblique aponeurosis, which is transpositioned into the preperitoneal space and sutured to the iliopubic tract. The details of this technique are herein described. After a follow-up ranging from 15 to 48 months, both the early and late complications are presented. They were minimal and of minor significance, apart from a hernial recurrence in one case. PMID- 9413056 TI - Orthotopic transplantation of a partial hepatic autograft in dogs. AB - The purpose of this study was to investigate the availability of an orthotopic transplantation of partial hepatic autograft in dogs as a means of surgical training. Male mongrel dogs weighting 10-15 kg were used. The left lobe of the liver was harvested while preserving the left branches of the portal vein, hepatic artery and bile duct, and the left hepatic vein. The remnant liver was removed while preserving the inferior vena cava using a veno-venous bypass. Orthotopic transplantation of the autograft was performed while anastomosing the left hepatic vein to the inferior vena cava, portal and arterial reconstruction, and external biliary drainage. Thirteen out of 29 dogs survived more than 48 h after transplantation. However, 6 out of 13 dogs were sacrificed after developing bile peritonitis due to a dislodgement of the biliary catheter, and only two dogs were able to survive for 7 days after transplantation. The arterial ketone body ratio recovered to 1.0 within 1 h after reperfusion, and the ratio of the dogs that survived for more than 48 h remained above 1.0 until sacrifice. Orthotopic transplantation of a partial hepatic autograft is a useful and simple procedure to train surgeons for partial liver transplantation. PMID- 9413055 TI - The effect of radioimmunotherapy using murine monoclonal antibody KIS1 on esophageal squamous cell carcinoma-bearing nude mice. AB - The monoclonal antibody (MoAb) KIS1 has been shown to react specifically with an antigen of human squamous cell carcinoma (SCC); however, a major problem in its clinical application is that the intact murine antibody induces a human anti mouse antibody (HAMA). To overcome this problem, we produced the KIS1 F(ab')2 fragment, then radioiodinated the intact KIS1 antibody and its F(ab')2 fragment. Nude mice bearing human esophageal SCC implants were injected with 100 microCi of 131I-intact KIS1 or 131I-KIS1 F(ab')2, and images were obtained using a gamma camera. Radioimmunotherapy (RIT) was performed by injecting the tumor-bearing nude mice with 131I-intact KIS1 or 131I-KIS1 F(ab')2 at a dosage of 300 microCi, following which 7 or 3 days were required to produce high quality tumor images by scintigraphy. The tumor-bearing mice treated with 131I-KIS1 F(ab')2 showed significant tumor growth inhibition, about 5.4 times greater than that of the control group and 1.8 times greater than that of the 131I-intact KIS1 group 21 days after the injection. These results indicate that the KIS1 F(ab')2 fragment is superior to intact KIS1, and that it may be clinically useful for radioimmunodetection followed by tumor targeting therapy for patients with SCC of the esophagus. PMID- 9413057 TI - Enhancing effect of thoraco-laparotomy on liver metastasis and the role played by active oxygens in its mechanism. AB - The enhancing effect on liver metastasis produced by the excessive surgical stress of thoraco-laparotomy (TL), and its regulation with a radical scavenger, were studied in 10-week-old Donryu rats. The rats were divided into three groups: those given thoraco-laparotomy for 1 h (the TL group); those given laparotomy alone for 1 h (the L group); those given a short laparotomy (the C group). The effects of treatment with 5 mg/kg of EPC-K1 was assessed in the TL group. A rat hepatocellular carcinoma cell line AH 60C (5 x 10(5) cells) was administered into the portal vein under general anesthesia. The number of metastatic liver nodules was counted 3 weeks later, and the lipid peroxide (LPO) levels of the liver and serum were measured by the TBA method on postoperative days (PODs) 1, 2, and 3. The number of metastatic liver nodules was 40.6 +/- 29.7, 15.0 +/- 15.8, and 13.7 +/- 9.4 in the TL, L, and C groups, respectively. When EPC-K1 was administered to the TL group, the LPO level on POD 1 decreased from 49.8 +/- 25.8 to 18.9 +/- 7.9 nM/g, and the number of metastatic liver nodules decreased from 27.2 +/- 30.0 to 8.9 +/- 12.7 in parallel. The findings of this study suggested that the excessive surgical stress produced by thoraco-laparotomy enhanced liver metastasis in parallel with an increase in LPO levels; however, the radical scavenger EPC-K1 could aid in reversing this effect. PMID- 9413059 TI - Anatomic segmental resection of the lung by thoracoscopy: an experimental study. AB - In patients who are unable to undergo a lobectomy for a small peripheral lung cancer, a partial thoracoscopic resection appears to be one viable alternative. However, since the regional lymphatics are disrupted in an anatomical fashion with a segmentectomy, it appears superior to a wedge resection. This experimental study was conducted to determine whether or not an anatomical segmental resection is feasible by thoracoscopy. A segmental resection of porcine lungs was performed using thoracoscopy. The segmental vessels were divided between ligatures. The segmental bronchus was divided by an endoscopic stapler. The intersegmental lung parenchyma was divided using a cotton dissector and a contact neodymium-yttrium aluminum garnet laser. Forty-three pigs were divided into seven groups as follows. Group 1: S1 + 2; group 2: S3; group 3: upper division; group 4: lower division; group 5: S6; group 6:S8; and group 7: S9 + 10. The operating times ranged from 145 +/- 15 min to 191 +/- 47 min. Blood loss ranged from 36 +/- 35 ml to 151 +/- 48 ml in all groups. The blood loss in the group with a resection of S6 and S9 + 10 was significantly greater than that of the other five groups. Most of the blood loss occurred during the division between the intersegmental planes. In conclusion, a thoracoscopic segmentectomy is considered to be technically feasible; however, further refinements in this technique are warranted before beginning clinical trials. PMID- 9413058 TI - The reinforcement of tracheoplasty with a self-fascia lata and Gelatin-Resorcin Formal (GRF) glue. AB - We examined the efficacy of protecting the suture line in tracheoplasty by using a self-fascia lata and Gelatin-Resorcin-Formal (GRF) glue. Fifteen dogs underwent a resection of four rings of the trachea and reconstruction, and we then observed them for one month; group A (n = 5) without reinforcement, group B (n = 5) with a self-fascia-lata spread with GRF glue, and group C (n = 5) with only a self fascia-lata. In the reinforced dogs (group B, three cases, and group C, five cases), in which the continuity of the suture line had been conserved, eight cases were resistant to pressures of from 240 mmHg to 300 mmHg, and two cases (both in group B) which had a partial discontinuity of the suture line were resistant to the same pressure of 160 mmHg. But in the five dogs without reinforcement (group A), four died from infection due to leakage of the trachea within 2 weeks; only one that had a continuity of the suture line survived and was resistant to pressure of 300 mmHg. These results show that a reinforcement of tracheoplasty using a self-fascia lata and GRF glue is useful for preventing air leakage from the suture line. PMID- 9413060 TI - Efficacy of prostaglandin I2 analog on liver grafts subjected to 30 minutes of warm ischemia. AB - Due to the fact that no effective conditioning agent for hemodynamically unstable donors exists, the number of suitable donors is limited. The efficacy of OP 41483, a stable analog of prostaglandin I2, as a conditioning agent of the liver was investigated in this study using pigs. OP-41483 was administered via the portal tributary for 10 min before 30 min of warm ischemia. Graft livers were procured after perfusion with OP-41483 in cold normal saline, then flushed with OP-41483 in Euro-Collins solution and placed in cold storage prior to orthotopic transplantation. OP-41483 was also administered intraportally for 120 min after reperfusion. Biochemical and histological studies, and survival rates were compared with a control group not given OP-41483 in an otherwise similar experimental protocol. The graft function recovered better in the OP group than in the control group, shown by the lactate values and lactate-to-pyruvate ratios. The marked congestion noted in the parenchyma of the control group livers was minimal in the OP group, verifying the microcirculatory effect of prostaglandin I2 by its vasodilatory and antithrombotic actions. These findings suggest that OP 41483 has some protective effect as a conditioning agent in liver transplantation, with timing of administration being crucial. PMID- 9413061 TI - Inflammatory fibroid polyp of the ileum with the appearance of a Borrmann type II lesion, caused by colostomy irrigation: report of a case. AB - Inflammatory fibroid polyps (IFPs) are rarely found in the gastrointestinal tract. The majority of IFPs are sessile-pedunculated or pedunculated polypoid lesions, whereas a polyp presenting like a Borrmann type II lesion is extremely unusual. This report describes the case of a 74-year-old man with a history of intussusception, in whom a preoperative diagnosis of a cecal tumor of the ileocecal valve was made. A laparotomy subsequently revealed a lesion similar to a Borrmann type II tumor located 15 cm above the ileocecal valve, but not at the valve. The lesion was diagnosed as an IFP which had been caused by repeated colostomy irrigation. The aim of the present report is to draw attention to this entity, which should be included in the differential diagnosis of intussusception and small bowel obstruction. PMID- 9413062 TI - A pancreatic pseudocyst with pancreatic pleural effusion: report of a case. AB - A 54-year-old man with a 30-year history of chronic alcoholism was admitted to our hospital suffering from dyspnea and left-sided chest pain. A chest radiograph revealed pleural effusion. Computed tomography revealed a pancreatic pseudocyst in the tail of the pancreas spreading out to the posterior mediastinum and the left pleural cavity. The laboratory findings of pleural effusion were as follows: amylase, 118,400 IU/1; protein, 4.6 g/dl; class I in cytology. Despite a reduction in the pleural effusion by conservative therapy, left back pain and a recurrence of the pleural effusion were observed after oral intake was re initiated. A distal pancreatectomy and ligation of the pancreaticopleural fistula were thus performed on the 75th hospital day. The patient made a complete recovery from pancreatic pleural effusion and has now been well for 9 years. PMID- 9413064 TI - A VATS lobectomy for lung cancer in a patient with an anomalous pulmonary vein: report of a case. AB - A video-assisted right upper lobectomy was successfully performed on a 58-year old man with an anomalous segmental pulmonary vein. The tumor was a peripherally located adenocarcinoma. The anomalous vein behind the right main bronchus was identified and safely divided. This case emphasized that to perform this procedure successfully, (1) a careful preoperative evaluation of the anatomy, including the presence of any possible vascular and/or bronchial anomalies, is necessary, and (2) if any anatomical structures cannot be determined intraoperatively, a conversion into an open procedure must immediately be undertaken. PMID- 9413063 TI - Nonoperative treatment for a ruptured pseudoaneurysm of the celiac trunk: report of a case. AB - We report the case of a 67-year-old man in whom hemorrhage from a ruptured celiac trunk pseudoaneurysm, which occurred as a consequence of leakage at the site of gastroduodenostomy, was successfully controlled by transcatheter arterial embolization (TAE) with stainless steel coils and N-butyl cyanoacrylate (NBCA). The occurrence of a pseudoaneurysm of the celiac trunk associated with anastomotic leakage is etiologically rare. We compiled reports from the literature on TAE for ruptured aneurysms of the celiac trunk, and compared its therapeutic value with that of surgical treatment. Operative death occurred in 4 of a series of 43 patients with aneurysms of the celiac trunk that were surgically treated (9.3%). In 5 patients with ruptured aneurysms, the operative mortality rate was 40% (2/5). Conversely, while the unsuccessful rate of TAE therapy was 17% (1/6), the mortality rate was nil. The patient whose case is presented here was affected by methicillin-resistant staphylococcus aureus (MRSA) at the site of leakage and in the lung. Under septic conditions such as hemorrhage secondary to pancreatitis, the mortality rate of surgical therapy was 23%-29%, whereas the success rate of TAE therapy was 79% and the mortality rate was 4%. Based on these findings, it is suggested that TAE therapy is a viable alternative to surgery for patients even with ruptured pseudoaneurysms of the celiac trunk. PMID- 9413065 TI - Squamous cell carcinoma arising from thyroglossal duct remnants: report of a case and results of immunohistochemical studies. AB - We report on the case of a 57-year-old male found to have squamous cell carcinoma (SCC) arising from thyroglossal duct remnants. The patient presented with an asymptomatic tumor in his anterior neck which was immovable on palpation. Aspiration biopsy cytology revealed class V malignancy with many atypical clusters and marked keratinization. After preoperative radiation, a radical operation employing Sistrunk's procedure with bilateral neck dissection was performed. Histopathological examination confirmed a diagnosis of moderately differentiated SCC, but revealed ciliated columnar epithelium in the walls of the cyst without a normal layer of squamous cells. Furthermore, immunohistochemical studies demonstrated the tumor to be negative for thyroglobulin staining, but positive for cytokeratin and carcinoembryonic antigen. These histopathological findings proved attributable to squamous metaplasia occurring in the ciliated columnar epithelium of the thyroglossal duct. Thus, SCC might originate in the metaplastic portion of the thyroglossal duct remnants. Although the prognosis associated with SCC in the thyroglossal duct is not as optimistic as that associated with papillary carcinoma, no evidence of recurrence has been observed in this patient in the 7 years since his operation. This suggests the effectiveness of our therapeutic approach for this unusual disease. PMID- 9413066 TI - Primary leiomyosarcoma of the breast. AB - Sarcomas of the breast are rare, accounting for about 1% of all malignant breast tumors. Leiomyosarcoma of the breast was an almost unknown tumor until some 20 years ago, and the few previously published cases lacked detailed information. Only 11 well-documented cases of leiomyosarcoma of the breast had been reported in the literature up to February 1992. The clinical features, diagnosis, therapy, and prognosis are discussed here in the light of the previously published literature. PMID- 9413067 TI - Wandering spleen in childhood: a report of three cases. AB - Wandering spleen is a rare cause of abdominal pain in children, and an accurate diagnosis is seldom made preoperatively. A splenectomy is the treatment of choice in cases of splenic torsion and infarction, while in patients with chronic symptoms splenopexy may also be attempted. We herein report three patients with wandering spleen, of whom two presented with acute torsion of the splenic pedicle and one demonstrated an asymptomatic mobile abdominal mass. In the first case splenopexy was attempted, but during follow-up the spleen was found to have undergone atrophy. The presentation, diagnostic procedures, and treatment modalities in pediatric wandering spleen are reviewed. PMID- 9413068 TI - A rat model with an isolated bladder in situ. AB - This paper describes our method for producing a rat model with an isolated bladder in situ in which the bladder makes no contact with urine. First, the right kidney was removed, then an external catheter was placed in the right ureter for bladder infusions, and next the left ureter was anatomosed to the proximal part of the descending colon. The animals were treated with antibiotics, and saline was infused daily into the bladder in order to prevent atrophy. This in situ model is considered to be useful in studies investigating the influence of specific compounds, such as carcinogens, on the bladder and its urothelium. PMID- 9413069 TI - Laparoscopic repair of a paraesophageal hiatus hernia without fundoplication. AB - We treated a case of paraesophageal hiatus hernia by laparoscopic repair. The procedure included a reduction of the gastric fundus and duodenal bulbus, closure of the diaphragmatic defect, mesh wrapping of the closure, gastropexy to the diaphragm, and a gastrostomy. Preoperative monitoring of the pH for 24 h showed no reflux. Intraoperative intraluminal manometry of the esophagus after hernia reduction showed the pressure of the lower esophageal sphincter to be normal, and thus an antireflux procedure was not deemed to be necessary. The patient was put on a soft diet from postoperative day 2. A postoperative upper gastrointestinal series showed no gastroesophageal reflux. No complications or recurrence of the hiatus hernia have been observed in the 12 months since the operation. Laparoscopic repair of a paraesophageal hiatus hernia with normal pressure of the lower esophageal sphincter, so that fundoplication is not needed, is thus considered to be possible. PMID- 9413070 TI - Fascia: an illustrative problem in international terminology. AB - As a result of international nomenclatures being in Latin, with the terms usually being undefined and translated into national vernacular languages, the same terms have been used in different ways in different countries. Fascia is an example of this, the limits of the meaning of the word differing in English-, French- and German-speaking countries. These differences are itemized in a comparative table. In 1989 the General Assembly of the International Federation of Associations of Anatomists [IFAA] created the Federative Committee on Anatomical Terminology [FCAT] with a remit to create a new terminology for the anatomical sciences with full democratic consultation with the member societies of IFAA. The draft recommendations of FCAT on fascia and the reasons for them are given. Problems associated with the use, or lack of use, of international terminologies are illustrated. For the advancement of medical knowledge and understanding, the cooperation of authors, editors and publishers in using the current terminology in the right way remains of critical importance. It is suggested that, after adopting a terminology, IFAA has a responsibility to arrange for technical back up. PMID- 9413071 TI - Morphologic characteristics of the fossa ovalis as an anatomic basis for transseptal catheterization. AB - Interest in transseptal catheterization has increased in recent decades, due to the fact that the technique has acquired a degree of security similar to other procedures. For a correct transseptal catheterization technique, it is essential to know the exact location of the fossa ovalis, its points of reference, and those morphologic parameters which allow us to assume precisely its dimensions. For this purpose, 97 human fresh hearts were studied (age range: 17-94 years; mean 63.74 +/- 18.50, 62 (64%) men and 35 (36%) women). The major and minor diameters of the fossa ovalis and the thickness of the intervenous tubercle were measured. In 22% of the cases, the fossa ovalis was permeable. Correlative tests between morphologic and general parameters were made. A positive correlation between cardiac weight and fossa ovalis area, and cardiac weight and thickness of the anterior limbus of the fossa was obtained. PMID- 9413072 TI - Anatomical basis for the choice of the femoral implant in the total hip arthroplasty. AB - There is within the population a considerable variation in the endosteal canal anatomy of the proximal extremity of the femur. This anatomical variation might jeopardize the result of the total hip arthroplasty when the surgeon uses a cementless femoral shaft. Indeed, if the secondary fixation of an implant stays under the dependency of many parameters, one has to obtain before anything else a very good primary stability, obtained by the morphological appropriateness of the bone-implant. The authors propose to define the cementless indications, as regards total hip arthroplasty, defining two radio-anatomical parameters: the Cortico-Medullary Index and the Femoral Flare Index. PMID- 9413073 TI - Basic anatomical investigation of semitendinosus and the long head of biceps femoris muscle for their possible use in electrically stimulated neosphincter formation. AB - Anal neosphincter formation with electrically stimulated gracilis muscle is used increasingly for the surgical treatment of fecal incontinence. An alternative to gracilis might be of interest if this muscle is not available. 30 semitendinosus muscles and 15 long heads of biceps femoris were investigated on human cadavers. In particular, the nerve and vascular supply of these muscles was studied, both representing basic factors for muscle transposition. The long head of biceps femoris m. was found to receive its dominant vascular supply from the first and second perforating artery and its nerve supply from one motor branch out of the sciatic nerve, both as described in literature. The examination of semitendinosus m., however, revealed new anatomical aspects in its vascular supply. In all cases semitendinosus m. was found to receive dominant vascular pedicles from the medial circumflex femoral artery close to the ischial tuberosity and the second perforating artery. The nerve supply consisted of two motor branches out of the sciatic nerve. Both muscles fulfilled several basic criterias for transposition to the anus. However, regarding these requirements, semitendinosus offered distinct advantages in comparison with the long head of biceps femoris. Due to its vascular and nerve topography, semitendinosus seems suitable to serve as an alternative to gracilis. PMID- 9413074 TI - Anatomic bases of the vascularized hepatic teres ligament flap. AB - The hepatic ligamentum teres was investigated in 30 adult specimens. The ligament represents a fibrous remnant of the umbilical v. and a small irregular lumen still exists in the ligament in adult life. The ligament is supplied by one set of independent round ligament a. and paraumbilical vv. Since the proximal segment of the ligament is easily mobilised and since the ligamentous a. originates from the right hepatic a. near the hilum, a proximal vascularized pedicle flap of the ligamentum teres has been recommended. Surgical repair of the extrahepatic bile duct using a vascularized pedicle flap of the ligamentum teres has been carried out successfully in 12 patients. PMID- 9413076 TI - Morphometry of the talocrural joint. AB - Based on two orthogonal radiologic views, the authors present a morphometric study of the talocrural joint. In 50 normal subjects, 10 parameters were measured and divided into 3 groups: the distal tibial joint surface parameters, the malleolar parameters and the talar parameters. These parameters were treated in both a descriptive and a correlative analysis. If the talocrural joint is a hinge joint whose talar articular surface can be simplified and classed as a cylinder segment, it is possible to calculate its curve radius. Then the correlative analysis allows to define the talar parameters and the corresponding parameters of the distal tibial joint surface. The malleolar parameters are independent factors. This study is the first morphologic analysis to serve as a basis for an ankle arthroplasty. PMID- 9413075 TI - The frontal sinus and the ethmoidal labyrinth. AB - In the "standard" anatomic description, the frontal bone and cribriform plate of the ethmoid bone form the base of the anterior cranial fossa. We studied the development of the ethmoidal bone as well as its relations to the frontal bone in macerated disarticulated skull bones and macerated skull bases of 35 individuals between 9 and 35 years of age. In 19 cases the ethmoidal cells were completely or partly uncovered by the frontal bone. In 6 of 19 cases the frontal bone did not cover any of the ethmoidal cells; in 10 further cases the frontal bone covered only the anterior and in 3 cases the anterior and middle ethmoidal cells. In a 60 year-old subject the ethmoidal cells were incorporated in the base of the anterior cranial fossa, a rare finding. Thus, a depressed lamina cribrosa is not the only danger in ethmoidectomy. Based on the present data ethmoidal cells uncovered by the frontal bone may involve a serious risk during ethmoidectomy even if the surgeon remains lateral to the insertion of the middle concha. The discrepancy between common descriptions of this region and our own findings may be related to imprecise data concerning the life stage of the cases described in the literature. PMID- 9413077 TI - The variational anatomy of the external aperture of the human vestibular aqueduct. AB - A study was undertaken to demonstrate the variational anatomy of the external aperture of the vestibular aqueduct in 90 human temporal bones obtained from 58 cadavers. Topographic landmarks of the posterior surface of the petrous bone are useful for general orientation and include the external aperture of the vestibular aqueduct, internal auditory meatus, sigmoid sinus, subarcuate fossa, superior petrosal sinus and cochlear canaliculus. We determined the mean distances from the external aperture of vestibular aqueduct to the above structures to be 10.98, 11.21, 9.42, 10.27 and 13.90 mm, respectively. Furthermore, the length of the external aperture of the vestibular aqueduct revealed significant differences between the right and left sides. The distances between the EAVA and certain anatomical structures on the posterior surface of the temporal bone should be taken into consideration during surgery. Knowing the variability of the position of the external aperture of the vestibular aqueduct may help surgeons avoid traumatizing, and thus producing inadvertent lesions to the hearing mechanism. PMID- 9413078 TI - An anatomic study of the lateral femoral cutaneous nerve. AB - In this study, the course of the lateral femoral cutaneous n. was examined bilaterally in 22 cadavers. Seven of these 44 lateral femoral cutaneous nn. showed variations in their course, especially in their number of branches under the inguinal ligament. During operations where the lateral femoral cutaneous n. may be demaged or in its decompression surgery, the nerve is to be found under the inguinal ligament. 1.52 +/- 0.84 cm medial to the anterior superior iliac spine. This nerve can be found passing through the inguinal ligament in as many as four branches. PMID- 9413079 TI - The marginal mandibular branch of the facial nerve. AB - The peripheral, extraparotid course and localisation of the marginal mandibular branch of the facial n. is described, with variations, based on the dissection of 40 cadaver half heads. Its anatomical relationships with the ramus of mandible and facial a. are studied and morphometric features are reported. Knowledge of the accurate course and relationship of the marginal mandibular branch should help to protect this nerve from surgical injury. PMID- 9413080 TI - Anatomy teaching: students' perceptions. AB - The goal of this study was to analyse students' perceptions of anatomy teaching. A questionnaire was distributed to two classes of first year dental students taught anatomy in both problem-based learning (PBL) and traditional formats. The questionnaire explored the students' most preferred techniques for learning anatomy, their examination preferences and their perceived level of mastery of anatomy. Fifty-seven (95%) students completed the survey. The most commonly used study aids were atlases, dissection and lecture notes (in descending order). Students expressed the desire for the final examination to include both written and oral components. Six months after the final examination, the students reported their perceived level of mastery of anatomy as either "very good" or "OK". Even in the PBL curriculum 39% of both classes felt it is necessary to have quizzes during the course to motivate and guide them in studying anatomy. PMID- 9413081 TI - In utero eyeball development study by magnetic resonance imaging. AB - The aim of this study was to measure fetal ocular development and to determine a growth curve by means of measurements in utero. Fetal ocular development was recorded by analysis of the results of magnetic resonance imaging (MRI). An anatomic study allowed definition of the best contrasted MRI sequences for calculation of the ocular surface. Biometric analysis of the values of the ocular surface in the neuro-ocular plane in 35 fetuses allowed establishment of a linear model of ocular growth curve in utero. Evaluation of ocular development may allow the detection and confirmation of malformational ocular anomalies such as microphthalmia. PMID- 9413082 TI - Radiologic anatomy of the metacarpophalangeal joints II to V. AB - The metacarpophalangeal (MCP) joints II to V of 21 hands were examined radiologically and arthrographically. Different recesses of the joint cavity were demonstrated both radiologically and macroscopically, with a dominating dorsal recess. The existing forms of the dorsal recess were one-tailed, two-tailed, three-tailed, symmetric and cap-like. Additionally, a palmar recess was found in the specimens examined, which presented as a small protrusion of the capsule and lay between the metacarpal head and the palmar plate. Furthermore, a distal recess was filled and unfolded in almost all the cases. Lateral recesses were found in the radial and ulnar directions beneath the collateral ligaments. The dorsal recess, due to its ability to collect fluid, is of clinical importance in pathologic processes causing effusions, while the clinical importance of the lateral recesses lie in their proximity to the stabilizing collateral ligaments of the metacarpophalangeal joints. The above mentioned recesses were seen as normal formations of the MCP joints and should therefore be taken into account in pathologic processes in this area. PMID- 9413083 TI - Determination of subacromial space width and inferior acromial mineralization by 3D CT. Preliminary data from patients with unilateral supraspinatus outlet syndrome. AB - A reduction of the subacromial space and an increased subacromial pressure have been considered to play an important role in the pathogenesis of rotator cuff lesions. The objective of the current study was to develop a CT based method for measuring the acromiohumeral distance and inferior acromial mineralization. In seven patients with unilateral rupture of the rotator cuff and two with impingement syndrome, transverse CT images were obtained at a section thickness of 1 mm with muscular relaxation in a standardized position. The bones were then reconstructed three-dimensionally, and the minimal vertical distance between the acromion and the humerus was determined in three secondary frontal images on both sides. The distribution of mineralization within the inferior surface of the acromion was assessed using CT osteoabsorptiometry. Although the Constant score was significantly reduced in the diseased shoulders, the width of the subacromial space was not routinely lower than on the contralateral side. In seven cases the maximal inferior acromial mineralization was identical in both shoulders, and in two cases it was lower on the affected side. These preliminary data suggest that with muscular relaxation no narrowing of the subacromial space can be detected in secondary frontal CT images, and that a potential increase of subacromial pressure is not high enough to cause a measurable increase in inferior acromial bone density. The method presented makes it possible to investigate the pathogenesis of the supraspinatus outlet syndrome in vivo with greater precision than has so far been possible with conventional radiography. PMID- 9413084 TI - Complete bipartition of the atlas in the Klippel-Feil syndrome. A radiologically illustrated case report. AB - A patient with the Klippel-Feil syndrome is described who presented with pain in the neck after a trauma. No fractures were found. Instead, a midline cleft in both the anterior and posterior atlantic arches was found which represents an extremely rare congenital anomaly. PMID- 9413085 TI - Determination of the position of the pelvic girdle using surface markers. AB - The overall movement of the trunk is made up of two components, namely the movements of the pelvic girdle and the vertebrae. In the frontal plane, the amplitude of the pelvic movements appears to be relatively limited compared to the vertebral column whereas the pelvis makes the major contribution to the total rotational movement in the axial plane. PMID- 9413086 TI - Efficacy of Salmonella typhi Vi capsular polysaccharide vaccine in South Africa. PMID- 9413087 TI - An immunogenicity and efficacy study of purified chick embryo cell culture rabies vaccine manufactured in Japan. AB - Purified chick embryo cell rabies vaccine manufactured by the Chemo-Sero Therapeutic Institute(Kaketsuken) at Kumamoto, Japan (Kaketsuken) was submitted to an immunogenicity and efficacy study. 52 severely rabies exposed patients were treated with the conventional five doses intramuscular WHO approved ('Essen') postexposure schedule. This included the administration of 40 IU kg-1 of equine rabies immune globulin on Day 0. A control group of equally severely exposed subjects were treated with human diploid cell rabies vaccine manufactured by the Swiss Serum and Vaccine Institute as well as human rabies immune globulin. There were no deaths in either group in the more than 2 years follow-up period. Subjects treated with the chick embryo vaccine showed greater suppression of the neutralizing antibody response by the equine rabies immune globulin than those given the human diploid cell vaccine and human rabies immune globulin. A group of 20 less severely rabies exposed patients who received only the chick embryo vaccine without immune globulin all had antibody titers greater than the WHO minimal acceptable level on Day 14, 30, 90 and 180. Fourteen subjects among the severely exposed vaccine and immune globulin study group were given vaccine boosters on Day 180 because of low antibody titers. It is concluded that chick embryo rabies vaccine manufactured by Kaketsuken is an immunogenic and effective rabies vaccine, but that the potency of future batches must be increased to provide a greater safety margin. PMID- 9413088 TI - Effects of carbohydrate modification of Quillaja saponaria Molina QH-B fraction on adjuvant activity, cholesterol-binding capacity and toxicity. AB - The iscom is an efficient antigen-presenting system for various antigens inducing both MHC class I and class II restricted immune responses. Protective immunity has been evoked against a variety of infectious agents. The saponin adjuvant Quil A, which was originally used to form iscoms, is composed of a mixture of structurally similar triterpenoids from Quillaja saponaria Molina having different biological activities. A purified, toxic Quillaja triterpenoid fraction with strong adjuvant activity, designated QH-B, was used to study whether modification of the carbohydrate moiety with sodium periodate would alter the toxicity without harming adjuvant activity and cholesterol-binding capacity. Most sugars, and in particular Api, Gal and Xyl, were modified by periodate treatment with only minor changes of the molecular weights indicating no loss of sugar residues. The adjuvant activity of QH-B was reduced in a dose-related manner, and at a concentration of 25 mM sodium periodate a significant reduction in toxicity was observed. The differences in both toxicity and adjuvant activity of the periodate-treated QH-B could be derived from alterations in the structure of the sugars Gal and Xyl, while modification of Api may influence adjuvant activity but not toxicity in vivo. The cholesterol-binding capacity, a prerequisite for iscom formation, was not affected by periodate oxidation at the doses tested. However, the use of modified QH-B as described in the present study for iscom-matrix formation resulted in "saponin-lipid complexes" which, to a various degree or totally, deviated from the characteristic iscom morphology. PMID- 9413089 TI - Protective immunity against heterologous challenge with encephalomyocarditis virus by VP1 DNA vaccination: effect of coinjection with a granulocyte-macrophage colony stimulating factor gene. AB - For DNA vaccination studies, recombinant VP1 protein of encephalomyocarditis virus (EMCV) was produced from Escherichia coli, and eukaryotic VP1 expression vector, pCT-Gs-VP1, was generated and used as a DNA vaccine. Mice were immunized intramuscularly (i.m.) with pCT-Gs-VP1 in the presence or absence of plasmid DNA expressing granulocyte-macrophage colony stimulating factor (GM-CSF), and were subsequently analyzed for their anti-VP1 immune responses with recombinant VP1 in ELISA. Immunization of mice with pCT-Gs-VP1 resulted in VP1-specific immune response and 43% protection from subsequent lethal heterologous challenge of EMCV. Coinjection of mice with pCT-Gs-VP1 and plasmid DNA encoding GM-CSF was shown to increase the seroconversion rate of the immunized mice with a single DNA injection, and enhanced to a higher degree VP1-specific immunity, which appeared to result in better protection (about 80%) from lethal virus challenge. Thus, our results provide evidence for the potential use of GM-CSF to induce better immune response and resistance against viral infection in DNA vaccination. PMID- 9413090 TI - Recombinant vaccine against hepatitis E: dose response and protection against heterologous challenge. AB - Thirty-two rhesus monkeys were used to evaluate the dose response of a recombinant HEV vaccine, and the efficacy of the vaccine based on the ORF2 protein of the Pakistani strain for pre- and post-exposure vaccination against intravenous challenge with homologous or heterologous virus was examined. Post exposure vaccination did not protect animals against hepatitis. Although primates vaccinated twice with 50-microgram, 10-microgram, 2-microgram, or 0.4-microgram doses of the recombinant 55 kDa ORF-2 protein were infected, they were protected from hepatitis when they were challenged with very high doses of the homologous strain of HEV. Primates vaccinated twice with a 50 micrograms dose of the recombinant protein were protected from hepatitis after heterologous challenge with the Mexican strain, the strain of HEV most genetically distant from the Pakistani strain. PMID- 9413091 TI - Chemical inactivation of recombinant vaccinia viruses and the effects on antigenicity and immunogenicity of recombinant simian immunodeficiency virus envelope glycoproteins. AB - The efficiency of paraformaldehyde (PFA) and binary ethylenimine (BEI) in inactivating recombinant vaccinia virus (rVV), present in baby hamster kidney cells expressing simian immunodeficiency virus envelope glycoproteins (SIV-Env), was measured in a series of inactivation studies. Both compounds were shown to be effective in reducing rVV titres. The use of standard 3-day titration assays proved to be inadequate to measure PFA inactivation, since upon prolonged incubation, residual rVV infectivity was detected in cultures negative at 3 days. Different procedures using PFA or BEI were selected to assess their influence on the antigenicity and immunogenicity or rVV expressed SIV-Env. Antigenicity, as defined by the ability to react with a panel of monoclonal antibodies recognizing major antigenic sites, and immunogenicity, as defined by the ability to induce SIV envelope specific and virus neutralizing serum antibodies in rats, proved to be preserved after either inactivation procedure. These data show that both protocols using PFA or BEI can be used successfully as part of the procedures to remove residual rVV infectivity. PMID- 9413092 TI - Expression of cereolysine AB genes in Bacillus anthracis vaccine strain ensures protection against experimental hemolytic anthrax infection. AB - The cereolysin AB genes from Bacillus cereus VKM-B164 have been expressed in Bacillus anthracis strains: virulent H-7 (PXO1, PXO2), vaccine STI-1 (PXO1), 221 (without its own plasmids). Expression was achieved by cloning the genes in a high copy number plasmid pE194. This construct was integrated with host genomes in amplified form. Gold hamsters were vaccinated with parental and recombinant B. anthracis STI-1 and 221 strains and challenged with virulent ones subcutaneously. Gold hamsters vaccinated with 221 strains showed, absence of protection. STI-1 immunisation protected against the H-7 strain, but did not protect against the recombinant strain. STI-1 recombinant strain protected gold hamsters against the H-7 as well as the recombinant H-7 strains. The results describe the modulation of immunopathogenic properties of B. anthracis due to expression of cereolysin AB genes. PMID- 9413093 TI - Nucleic acid vaccination of Brucella abortus ribosomal L7/L12 gene elicits immune response. AB - Nucleic acid vaccines provide an exciting approach for antigen presentation to the immune system. As a test of this new methodology, the immune response to the in vivo-expressed Brucella abortus ribosomal L7/12 gene in the muscle cells of mice was examined. To accomplish this goal the eukaryotic expression systems pcDNA3 and p6 were used. Single intramuscular injection of the L7/L12 gene driven by the human cytomegalovirus (CMV) promoter (pcDNA3) or bovine MHC 1 promoter (p6) resulted in intracellular expression of the B. abortus L7/L12 immunodominant protein encoded by this gene. This application facilitated directed antigen presentation to the immune system and established specific antibody and T-cell responses compared with vector only (pcDNA3) negative controls and B. abortus S19 injected positive controls. Although pcDNA3-encoded L7/L12 gene-inoculated mice possessed significant protection, p6-L7/L12 did not engender significant protection against B. abortus S2308 infection compared to positive control mice. These data suggest a promising antigen-specific response, and L7/L12 nucleic acid vaccination may be an initial step in the development of genetically engineered candidate vaccines against brucellosis. This study for the first time focuses on DNA immunization of a gene from B. abortus. PMID- 9413094 TI - Comparative efficacy of two immunocontraceptive vaccines. AB - As part of our research program to develop immunocontraception as a wildlife management tool, we compared the physiological responses of wild Norway rats (Rattus norvegicus) to two immunocontraceptive vaccines; one involved mouse zona pellucida peptide (MZPP); the other involved gonadotropin releasing hormone (GnRH). Efficacy was monitored by immune, hormonal, and natality responses. Both vaccines were effective, but GnRH was much more effective (100% sterility of both sexes vs. 50% sterility of MZPP-treated females). Breeding success of control rats was 88% with litters of 5-9 pups; breeding success of MZPP rats was 50% with litters of 2-8; GnRH rats produced no young. In GnRH-treated male rats monitored for up to 17 months, testosterone was nondetectable and testes were atrophied to about 10% of their original volume for 10-13 months. There were no notable differences in mortality or body weights among groups, and, with the exception of testicular regression, there were no changes in general appearance. The GnRH vaccine is potentially a good rat reproductive control agent that may be effective over the normal lifespan of a rat under natural conditions in the wild. PMID- 9413095 TI - TgPVR21 mice for testing type-3 oral poliovirus vaccines: role of clinical observation and histological examination. AB - Transgenic mice susceptible to poliovirus (TgPVR mice) have been used to study poliovirus neurovirulence and attenuation. It was shown recently that mouse line TgPVR21 may be a suitable model to evaluate neurovirulence safety of oral poliovirus vaccine. It was important to determine whether TgPVR21 mice are sensitive enough to discriminate between type-3 reference and 'marginal' vaccines, i.e. those that failed the monkey test while containing only slightly increased amounts of neurovirulent revertants at position 472 of the viral genome as measured by a molecular assay MAPREC. Data presented here demonstrate that TgPVR21 mice are not less sensitive than monkeys in the detection of marginal vaccines. In contrast to the monkey neurovirulence test, which is based on histological examination of the CNS, the TgPVR21 mouse neurovirulence test revealed marginal vaccines by simple analysis of clinical signs without requiring a laborious histological examination. PMID- 9413096 TI - Long-term high immune response to diphtheria toxoid in rodents with diphtheria toxoid conjugated to dextran as a single contact point delivery system. AB - Cross-linked dextran beads were used as a carrier for development of a 'single contact' vaccine delivery system. Diphtheria toxoid (DT) was covalently coupled to dextran beads (DEX-DT conjugate). The conjugate was immunoreactive with antibodies raised against native DT. Immunization of rats with DEX-DT conjugate generated on average 24 times higher antibody titres against diphtheria toxoid than immunization with the conventional DT absorbed on alum. The immune response was sustained for 9 months, with gradual decline of antibody titres thereafter. Significant antibody titres were measurable in rats after 1 year of immunization. DEX-DT had neither acute nor long-term adverse effects on the health of animals, as evidenced by local reactions, general behaviour, food intake, body weight gain and survival compared with controls. PMID- 9413097 TI - The outer surface protein A (OspA) vaccine against Lyme disease: efficacy in the rhesus monkey. AB - The efficacy of an outer surface protein A (OspA) vaccine in three different formulations was investigated in the rhesus monkey. The challenge infection was administered using Ixodes scapularis ticks that were infected with the B31 strain of Borrelia burgdorferi. Protection was assessed against both infection and disease, by a variety of procedures. Some of the animals were radically immune suppressed, as an attempt to reveal any putative low level infection in the vaccinated animals. The significant difference found between the spirochaetal infection rates of ticks that had fed on vaccinated vs. control monkeys, lack of seroconversion in the vaccinated animals, and the absence of spirochaetal DNA in the skin of vaccinated animals in the weeks following the challenge, indicate that vaccinated monkeys were protected against tick challenge. The post-mortem immunohistochemical and polymerase chain reaction analyses, however, suggest that these monkeys may have undergone a low-level infection that was transient. PMID- 9413098 TI - The immune response to a model antigen associated with PLG microparticles prepared using different surfactants. AB - The effect of different surfactants on the surface characteristics of poly(D,L lactideco-glycolide) microparticles prepared by the emulsification/solvent evaporation technique was investigated and the immune response to a protein antigen (OVA) associated with these microparticles was measured. Three surfactants--polyvinyl alcohol (PVA, a conventional stabilizer of PLG microparticles), the non-ionic surfactant, poly(oxyethylene glycerol mono-oleate) [Tagat] and Bile salts (a natural emulsifer)--were used to produce OVA-loaded PLG microparticles. Antigen was detected at the surface of all three types of OVA loaded microparticles, in amounts in excess of 40% of the total protein load. The levels of specific serum IgG antibody elicited to OVA were significantly higher (P < 0.05) after a single subcutaneous administration of antigen associated with the Bile salts and Tagat formulations compared to the PVA formulation. A strong correlation was revealed between the levels of antibody measured and the magnitude of negative surface charge of the particulate carrier. The pattern of the IgG antibody response to OVA was similar in all three cases, indicating that the degradation rate of the PLG polymer determined the duration of the response. The results demonstrate the potential of using different surfactants to produce PLG microparticles with increased adjuvant activity. PMID- 9413099 TI - Seroconversion of a trivalent measles, mumps, and rubella vaccine in children aged 9 and 15 months. AB - The serological response to MMR vaccine was evaluated in 109 9-month-old infants having no history of measles vaccination, and in 98 15-month-old children who had received monocomponent measles immunisation at 9 months. The combined vaccine contained Schwarz, Urabe Am9, and Wistar RA 27/3 live attenuated virus strains. Preimmunisation antibody levels were extremely low for the 9-month-old children, indicating that maternally-transmitted antibodies do not persist at this age. In the case of mumps, preimmunisation antibody levels were significantly higher in the 15-month-old than in the 9-month-old group. A difference between groups in terms of postimmunisation antibody titres was observed only for rubella, with titres being significantly higher in the older group. Seroconversion rates were high in both groups and no serious events attributable to vaccination were observed. The MMR vaccine can thus be administered to children as young as 9 months of age. Evidence for the efficacy of a two-dose schedule, i.e. at 9 and 15 months, is presented. PMID- 9413100 TI - A comparison of antibody responses to veterinary vaccine antigens potentiated by different adjuvants. AB - Six adjuvant formulations were compared for their ability to potentiate the primary and memory antibody responses in mice to three companion animal vaccine immunogens--feline leukemia virus (FeLV), feline immunodeficiency virus (FIV), and a recombinantly-derived heartworm antigen. The combination of a novel bacterial immunostimulator, gliding bacterial adjuvant (GBA), either adsorbed onto an aluminum hydroxide gel (Rehydragel HPA), or emulsified with a vehicle of polyalcohol and detergent, elicited the strongest memory responses to both virus preparations. Both forms of aluminum hydroxide gels administered without GBA gave similar levels of adjuvant effects, on par with or greater than those generated by incomplete Freund's adjuvant (IFA). The Acemannan immunostimulant was not effective in increasing the responses to the virus antigens, but increased the primary response to the heart-worm antigen over tenfold from control levels. All preparations appeared to be well tolerated, with no detectable adverse reactions observed in any of the 250 mice used. The proven safety of aluminum hydroxide adjuvants and the apparent absence of adverse reactions seen with GBA make this vehicle/adjuvant formulation worthy of additional study. PMID- 9413101 TI - Immunization of cattle with a BHV1 vector vaccine or a DNA vaccine both coding for the G protein of BRSV. AB - A gE-negative bovine herpesvirus 1 (BHV1) vector vaccine carrying a gene coding for the G protein of bovine respiratory syncytial virus (BRSV) (BHV1/BRSV-G) induced the same high degree of protection in calves against BRSV infection and BHV1 infection as a multivalent commercial vaccine. A DNA plasmid vaccine, carrying the same gene as the BHV1/BRSV-G vaccine, significantly reduced BRSV shedding after BRSV infection compared with that in control calves, but less well than the BHV1/BRSV-G vaccine. Flow cytometric analysis showed a significant relative increase of gamma/delta+ T cells in peripheral blood after BRSV challenge-infection of the calves of the control group but not in the vaccinated groups. These results indicate that the G protein of BRSV can induce significant protection against BRSV infection in cattle, and that the BHV1/BRSV-G vaccine protects effectively against a subsequent BRSV and BHV1 infection. PMID- 9413102 TI - Specific immune response in adult medical personnel immunized with acellular pertussis vaccine with special emphasis on T helper cell response. AB - Recent epidemiologic data have indicated that adults are the most important reservoir that transmit pertussis to children. However, conventional whole cell pertussis vaccine is contraindicated in adults and children over 7 years of age because of the unacceptably high rate of adverse reactions. The aim of this study is to evaluate the specific cellular immune responses and adverse reactions to a less reactogenic acellular pertussis vaccine in adult volunteers. Eighty healthy medical personnel in Chang Gung Children's Hospital were enrolled. Volunteers in each group received: (1) Td + full strength acellular pertussis vaccine (PT, 1 microgram/0.5 ml; FHA, 4 micrograms/0.5 ml); (2) Td + half strength acellular pertussis vaccine; (3) Td alone. Lymphocyte phenotypic analysis, antigen-specific antibody titers, antigen-specific proliferative response and cytokine levels were evaluated before and 1 month after vaccination. Our data revealed: (1) the adverse reactions were minimal; (2) phenotypic analysis showed no non-specific activation of helper T or memory T cell after vaccination; (3) both PT and FHA specific antibody titers increased significantly after vaccination, (4) PT antigens had a mitogenic effect on cord blood mononuclear cells and peripheral blood mononuclear cells of the adult volunteers; (5) FHA-specific T cell proliferative responses significantly increased after vaccination; (6) the cytokine production pattern showed predominant activation of Th 1 cells as reflected in increased production of gamma-IFN after vaccination. Acellular pertussis vaccine can effectively induce both humoral and cellular immune response in adults. PMID- 9413103 TI - Campylobacter jejuni in broiler chickens: colonization and humoral immunity following oral vaccination and experimental infection. AB - A formalin inactivated, Campylobacter jejuni whole cell vaccine, either with or without Escherichia coli heat labile toxin (LT) as a mucosal adjuvant, was administered orally to broiler chickens. Three vaccine trials were performed, differing in the number of vaccinations, and time of administration, as well as the inclusion and dose of LT. The overall reductions of C. jejuni colonization in the vaccinated chickens ranged from 16 to 93% compared with non-vaccinated controls. Enhanced levels of anti-C. jejuni secretory IgA antibodies were demonstrated in vaccinated chickens. Vaccination also appeared to induce an anamnestic response to C. jejuni antigens in the 14-33 kDa range, as demonstrated by Western immunoblots. Interestingly, the inclusion of LT in the vaccine regimen did not appear to boost the immunogenicity of the vaccine. These results are encouraging and suggest that future development of successful oral vaccines for the control of enteropathogenic Campylobacter in poultry is feasible. PMID- 9413104 TI - Liposomally-encapsulated ricin toxoid vaccine delivered intratracheally elicits a good immune response and protects against a lethal pulmonary dose of ricin toxin. AB - A small study was performed to examine whether the instillation of ricin toxoid vaccine into the lungs of Porton rats offered protection from lethal effects of subsequent intratracheal challenge with ricin toxin. Further the immune response to liposomally-encapsulated vaccine and the protection offered was compared with vaccine either adsorbed to Alhydrogel adjuvant or as a simple aqueous solution. The formaldehyde-treated ricin toxin vaccine (RTV) was administered at two dose levels, 500 and 100 micrograms kg-1 body weight to groups of rats, on two occasions by intratracheal instillation. Liposomally-encapsulated vaccine (LRTV) produced a higher titre of ricin-specific antibodies than Alhydrogel-vaccine (ARTV) and vaccine solution. When challenged with 3 LD50 of ricin by intratracheal instillation 7 weeks after the second vaccine instillation, all rats in both LRTV dose groups survived with minimal signs of incapacitation. Analysis of antibody secretion by spleen cells, 14 days post challenge, showed that the IgG isotype in the LRTV group was significantly higher than that in the ARTV and RTV groups and also that the proportion of specific IgA in lung fluid was higher in the LRTV group than in the ARTV and RTV groups. The results of this study indicate that effective vaccinations against inhaled ricin could be achieved with liposomally-encapsulated ricin toxoid, via the lung and should be investigated further. PMID- 9413105 TI - A subunit vaccine based on glycoprotein E2 of bovine virus diarrhea virus induces fetal protection in sheep against homologous challenge. AB - The primary aim of a bovine virus diarrhea virus (BVDV) vaccine is to prevent transplacental transmission of virus. E2 genes of three BVDV strains, belonging to antigenic groups IA, IB and II, were expressed in insect cells. Three groups of 12 ewes were immunized twice with one of the E2 proteins. A fourth group served as a control. The ewes were served and the pregnant ewes of each vaccination group were allotted to three different challenge groups. Seven weeks after the second vaccination the ewes were challenged intranasally with one of the three BVDV strains. Three weeks later the fetuses were removed and fetal organs were collected for virus isolation. At the day of challenge all vaccinated ewes had neutralizing antibodies against the homologous BVDV strains. One E2 subunit vaccine prevented fetal infection after homologous challenge. PMID- 9413106 TI - Affinity-purified dengue-2 virus envelope glycoprotein induces neutralizing antibodies and protective immunity in mice. AB - We constructed a recombinant baculovirus which produces a dengue (DEN)-2 virus envelope (E) protein containing a six-histidine (H6) tag in place of the last 100 amino acids at its C-terminus. The recombinant protein was purified from the supernatant of baculovirus-infected Spodoptera frugiperda insect cell cultures to apparent homogeneity by cation-chelation chromatography (TALON) in which the H6 tagged E-protein was eluted under non-denaturing conditions with 100 mM imidazole at pH 8.0. Mice vaccinated with the purified E mixed with aluminium hydroxide adjuvant showed an immune response of IgM and IgG1, IgG2a and IgG2b isotypes, and neutralizing antibodies, similar to that following immunization with purified inactivated DEN-2 virus. Moreover, mice that received the purified recombinant protein were significantly protected against 200 50% lethal dose of DEN-2 virus. This combination of affinity purified H6-tagged protein and adsorption onto a cationic carrier seems promising for the production of immunogenic particulate proteins, especially DEN protein for the four serotypes, and the development of a new generation of vaccines. PMID- 9413107 TI - Two primary doses of diphtheria-tetanus-acellular pertussis vaccine induce immunological responses to Bordetella pertussis as strong as those induced by three primary doses. AB - Pertussis vaccinations are administered worldwide under various conditions and schedules with diphtheria-tetanus-pertussis (DTP). In Japan, a general vaccination with three primary doses of diphtheria-tetanus-acellular pertussis (DTaP) at 4-week intervals and one booster dose 12 months after the primary series have been used since 1981. Decreasing the number of doses of the vaccination would lessen the physical and economic costs. To compare the immunological response to two versus three primary doses, we assessed antibody and cellular immune responses in health children. The anti-filamentous hemagglutinin (anti-FHA) and anti-pertussis toxin (anti-PT) antibody responses to two primary doses of DTaP before a booster were significantly lower than the responses to three primary doses. Although these antibody levels were low in children who received two primary doses, the FHA-induced DNA synthesis was equal to that of the children who received three doses. The anti-FHA and anti-PT antibody levels 4 weeks after the booster following two doses were similar to the levels following three doses, and high antibody titers were maintained over a long period. In areas where contact with bacteria is expected, two primary doses of DTaP may be adequate to induce the necessary level of immunological responses. PMID- 9413108 TI - Aqueous-based microcapsules are detected primarily in gut-associated dendritic cells after oral inoculation of mice. AB - We previously found that aqueous-based microencapsulation enhanced virus-specific humoral immune responses after oral inoculation of mice. However, the mechanism by which microencapsulation enhances immunogenicity remains unclear. We found that spermine-alginate microcapsules were detected primarily in gut-associated dendritic cells (i.e. CD11c/CD18+, Ia+, CD11b-, CD45R-) after oral inoculation of adult mice. Microencapsulation may enhance immunogenicity by involving antigen presenting cells which are more efficient than those recruited during natural infection. PMID- 9413109 TI - Age-related differences in norfloxacin pharmacokinetic behaviour following intravenous and oral administration in sheep. AB - The pharmacokinetics of norfloxacin after intravenous (i.v.) and oral (PO) administration in lambs (n = 5) and adult sheep (n = 5) were studied. After i.v. administration (10 mg.kg-1) plasma concentrations were best fitted by a three compartment open model in both age groups. Distribution volumes were significantly larger in lambs (approximate 4.0 fold difference between 4 week old and adult sheep). There was no significant difference (p < 0.05) between the groups in terms of elimination halflife but plasma clearance was significantly higher in lambs. Norfloxacin was poorly absorbed after oral administration (60 mg.kg-1) in sheep (F = 4.04%). Mean oral bioavailability was 73.51% in lambs (30 mg.kg-1). Norfloxacin elimination was faster in lambs after oral administration. MRTt was significantly prolonged in both age groups when compared with the respective data for i.v. administration. PMID- 9413110 TI - The intake of polyunsaturated fatty acids by cats is reflected in their adipose tissue. AB - To find out whether the composition of the subcutaneous adipose tissue of cats reflects the intake of polyunsaturated fatty acids, we performed a feeding trial. Six groups of kittens were fed on diets with variable combinations of corn, linseed, and fish oil. After 5 months, biopsies of subcutaneous adipose tissue were analysed for their contents of linoleic, alpha-linolenic, eicosapentaenoic, and docosahexaenoic acid. The observed strong correlations between dietary and fat tissue polyunsaturated fatty acids indicate that the fatty acid composition of adipose tissue may be used as an index of the fatty acid composition of the diet. Thus, in epidemiological studies on the possible relationship between dietary fat type and feline disease the fatty acid composition of adipose tissue might be a useful measure. PMID- 9413111 TI - Airborne dust and aeroallergen concentrations in different sources of feed and bedding for horses. AB - Standardized methods were used to make quantitative and qualitative assessments of respirable dust and aeroallergens in feed and bedding for horses. Concentrations of airborne dust were measured by using a Rion particle counter, and levels of major aeroallergens implicated in chronic obstructive pulmonary disease were measured by using an Andersen sampler. Laboratory conditions allowed comparison of the different sources of forage, supplements, and bedding without external influences such as ventilation, external temperature and horse activity affecting the result. Grass silages of approximately 50% dry matter and alfalfa pellets appeared to be very good sources of forage with low levels of dust and aeroallergens. The studied good quality straw was significantly less dusty with fewer allergens than the wood shavings. Supplements, such as whole grains and molassed concentrates, contained many respirable particles and aeroallergens. Rolled grains were significantly more dusty than good hay. PMID- 9413112 TI - Comparison of air- and bone-conducted brain stem auditory evoked responses in young dogs and dogs with bilateral ear canal obstruction. AB - Brain stem responses to air- and bone-conducted stimuli were analyzed in 11 young dogs, using an in-the-ear transducer and a vibrator designed for human hearing tests, respectively. The mean thresholds were 0 to 10 dB for air-conducted stimuli and 50 to 60 dB for bone-conducted stimuli. The wave forms and inter-peak latencies of the waves of the auditory evoked responses elicited by air-conducted and bone-conducted stimuli were similar. This indicated that the signals had the same origin and thus both the air-conducted and the bone-conducted responses could be considered to be auditory responses. Measurement of air-conducted and bone-conducted brain stem-evoked responses in five dogs with bilateral chronic obstructive ear disease revealed thresholds of 50 to 60 dB for air-conducted stimuli and 60 to 70 dB for bone-conducted stimuli. By comparison of these results with those in the 11 young dogs, it could be concluded that there was hearing loss other than that caused by obstruction of the ear canals. PMID- 9413113 TI - Limited capacity of neonatal rabbits to eliminate enrofloxacin and ciprofloxacin. AB - The pharmacokinetics of enrofloxacin (ENR) and ciprofloxacin (CIP) in newborn and young rabbits were studied. Rabbits of different ages (1-, 8-, 16-, and 30-day old) were administered, by the intraperitoneal route (i.p.), a dose of 7.5 mg of either drug/kg. In 1-, 8-, and 16-day-old rabbits, blood samples were drawn by cardiac puncture, under light ether anaesthesia, at predetermined times after drug administration. In 30-day-old rabbits, serial blood samples were drawn through an arterial catheter. Plasma was immediately obtained and analysed using an HPLC method. ENR and CIP plasma protein binding was also determined. The plasma pharmacokinetic profiles of ENR and CIP obtained for 30-day-old rabbits agreed with those reported in the literature for healthy adult rabbits. Nevertheless, significant differences were observed for the body clearance, the slope of the terminal phase, the volume of distribution, and the area under the curve when compared with those for younger animals (1-, 8-, and 16-day-old rabbits), indicating a limited capacity of neonatal rabbits to eliminate ENR and CIP. No differences were found when we compared the calculated values for ENR or CIP plasma protein binding as a function of the postnatal age, indicating that development does not seem to alter the free fraction of these drugs in the rabbit. Taking into account that extensive placental and milk transfer has been reported for these drugs after administration to pregnant or nursing rabbits, a cautious, attitude regarding their use in these animals must be adopted. PMID- 9413114 TI - A survey of anthelmintic resistance in nematodes of sheep in The Netherlands. AB - The prevalence of anthelmintic resistance in nematodes of sheep was surveyed in 1994 on 70 farms in the Netherlands. An in vitro egg hatch assay, faecal egg count reduction (FECR) 14 days after treatment, and larval cultures were used as methods of investigation. Oxfendazole was tested on 69, ivermectin on 51, and levamisole on 36 farms. The median effective dose (ED50) of thiabendazole could be determined on 64 farms. On 60 farms (94%) the ED50 value was > or = 0.12 microgram ml-1, which is indicative of the presence of benzimidazole (BZ) resistance. On two farms egg output was too low to do a FECR test. Based on the results of the FECR test, BZ resistance was present on 56 farms (84%), on 2 farms there was a suspicion of resistance and on 9 farms no resistance could be found. No clear indications were found for the presence of resistance against ivermectin or levamisole. BZ resistance was demonstrated in Haemonchus contortus, Cooperia curticei, Ostertagia spp. and/or Trichostrongylus spp. No resistance was observed in species from the genus Nematodirus, Chabertia ovina and/or Oesophagostomum spp. PMID- 9413115 TI - Xanthinuria in a family of Cavalier King Charles spaniels. AB - Xanthine calculi were found in a 7-month-old male Cavalier King Charles spaniel with urethral obstruction and renal insufficiency. Because the only two other reported cases of naturally occurring xanthine urolithiasis concerned a Cavalier King Charles and a King Charles spaniel the urine of the littermates and parents of the patient were also examined for xanthinuria. Semi-quantitative analysis revealed high urine concentrations of hypoxanthine and xanthine in the patient and his female littermate. Quantitative analysis by high-pressure liquid chromatography (HPLC) of the urine samples from the family of this Cavalier King Charles spaniel and nine control dogs revealed that hypoxanthine and xanthine excretion was 30 and 60 times higher in the affected patient and the female littermate than in the others dogs. The pattern of xanthinuria, which is caused by a deficiency of the enzyme xanthine oxidase, in the relation diagram of this family of Cavalier King Charles Spaniels was consistent with an autosomal recessive mode of inheritance. PMID- 9413116 TI - The water-holding capacity of fresh meat. AB - Several aspects of the water-holding capacity of fresh meat are discussed. After some compositional and structural characteristics of muscle are outlined, special attention is paid to post mortem muscle physiology and related mechanisms of fluid loss from meat. Finally, determinants of water-holding capacity (physiological factors, rearing conditions and processing factors) are described. From this review it can be concluded that the water-holding capacity and the subsequent drip loss are complex attributes of meat which are influenced by factors in the whole meat production chain. However, the effects of many factors, such as rearing conditions, administration of growth promoters, feed, and ageing of meat, are still not clear. To be able to optimize and control the water holding capacity of meat, it is important to have information about the effects of each processing factor and the basic mechanisms involved. Once these mechanisms are known, the influence of biological and technological factors will be more predictable and more easy to control. Veterinarians can play an important role in this. PMID- 9413117 TI - A case of Ehlers-Danlos-like syndrome in a rabbit with a review of the disease in other species. AB - A case of marked skin fragility in a 4-month-old pet rabbit is described. The clinical findings, gross pathology, histopathology, and ultrastructure of skin samples were consistent with Ehlers-Danlos-like syndrome. This syndrome is recognized in many animal species and is often compared to Ehlers-Danlos syndrome in humans. Ehlers-Danlos-like syndromes in animals are reviewed and possible similarities between these disorders and Ehlers-Danlos syndrome in humans are discussed. PMID- 9413118 TI - Deacetylation as a determinant of sulphonamide pharmacokinetics in pigs. AB - Sulphamonomethoxine (SMM), sulphadimidine (SDD), sulphadiazine (SDZ) and their N4 acetyl derivatives (AcSMM, AcSDD and AcSDZ) were intravenously injected into Goettingen miniature pigs and deacetylation was evaluated from plasma concentration-time curves, renal excretion, and rate constants obtained from pharmacokinetic analysis, using a non-linear least-squares method. Deacetylated metabolite was detected in both plasma and urine after intravenous injection of AcSMM, AcSDD and AcSDZ. The area under the curve (AUC) values for the deacetylated metabolite were significantly higher than those for acetyl derivatives after AcSMM and AcSDD administration, but significantly lower after AcSDZ. After AcSMM and AcSDD injection, the concentration ratio between deacetylated metabolite and acetyl derivative was almost constant in the terminal linear phase and similar to that seen after injection of sulphonamide. After AcSDZ injection, however, a constant ratio was not observed. These results indicate that deacetylation can have a significant effect on the pharmacokinetics of SMM and SDD, but not on those of SDZ in pigs. The rate constant for deacetylation was significantly higher than that for acetylation for SMM and SDD, but significantly lower for SDZ. It is, therefore, concluded that deacetylation may be a determinant of the pharmacokinetics of SMM and SDD in pigs. It was, however, not a determinant of SDZ pharmacokinetics because N4-acetylation is not the main elimination route in pigs. PMID- 9413119 TI - Treatment of isolated tibial fractures in cats and dogs. AB - We report here a review of 33 cases of isolated tibial fractures (i.e., with the fibula intact) in 10 cats and 23 dogs, presented to four orthopaedic referral clinics. The purpose of this study was to identify factors to be considered when selecting the therapy for an isolated tibial fracture in cats and dogs. The animal species, the size of the dog breed, the age of the patient, the type of tibial fracture, the presence of an additional fracture, the treating clinic, the therapy applied, and the results after treatment were taken into account. A step backward logistic regression analysis was applied to the series of cases to examine possible relations among the covariates. Treatment outcome was found by logistic regression analysis to depend significantly (P < or = 0.05) on age, the presence of an additional fracture, and the therapy applied. No other relations were present between the covariates. In the immature cat or dog with an isolated tibial fracture, treatment by external splinting has a good prognosis. In the mature cat or dog with an isolated tibial fracture, rigid fixation by external fixator or internal fixation is the method of choice. PMID- 9413120 TI - Behavioural signs of oestrus during pregnancy in lactating dairy cows. AB - Two herds of approximately 50 dairy cows were observed for oestrous behaviour for 6 weeks, 12 time a day for 30 minutes. It appeared that 3.08% of the pregnant cows showed oestrous behaviour during pregnancy (EBP) in such an intensity that they would have been considered in oestrus. With a less rigid oestrus detection threshold, 10.8% of the pregnant cows would have been considered to be in oestrus. Animals showed EBP during all months of pregnancy, but most of the behavior was observed in the middle of the gestation period. PMID- 9413121 TI - A retrospective clinical study of canine leishmaniasis in 150 dogs naturally infected by Leishmania infantum. AB - The clinical and laboratory findings observed in 150 dogs naturally infected by Leishmania infantum, from a large endemic area of southern Italy, are described. There was a gradual onset of clinical signs and the course of the disease was progressive in almost all the cases. The majority of the dogs were mongrels (43.3 per cent), male (64.7 per cent), of medium size (50.6 per cent), three to seven years old (64.7 per cent), and living outdoors (60 per cent). They showed generalised (56.7 per cent) or symmetrical (32 per cent) lymphadenomegaly; the mucous membranes of 87 of the dogs (58 per cent) were pale and moderate or severe splenomegaly was diagnosed in 80 dogs (53.3 per cent); weight loss was observed in 32 per cent of the animals. Skin abnormalities were very common, and included dry exfoliative dermatitis (56 per cent), ulcers (40 per cent) periorbital alopecia ('lunettes') (18 per cent), diffuse alopecia (14 per cent) and onychogryphosis (24 per cent). Ocular signs were observed in 24 dogs (16 per cent) including 16 cases of keratoconjunctivitis (three with keratoconjunctivitis sicca), six cases of moderate uveitis and two cases of panophthalmitis. The acute form of the disease was diagnosed in only six dogs and was characterised by fever and generalised lymphadenomegaly, and by the absence of skin lesions. Another six dogs had severe renal failure without systemic clinical signs of leishmaniasis. The most important laboratory findings were a severe or moderate increase in gammaglobulins, hypoalbuminaemia, hyperproteinemia and anaemia. Cultures or cytology tests for L infantum parasites were positive in 134 of the dogs. Following the standard procedures developed for human lymph node and bone marrow cytology tests, the leishmania density in the dogs varied from 1+ to 2+. Leishmania antibody titres were high (> 1:160) in almost all the dogs. Immunological tests for autoantibodies were positive in 25 of 53 dogs tested in the antinuclear antibody (ANA) test, in 15 of 43 dogs tested in the latex test and in five of 24 dogs tested in the Coombs test. PMID- 9413122 TI - Experimental reproduction of iodine deficiency in cattle. AB - The role of iodine deficiency in stillbirth/perinatal weak calf syndrome was investigated in pregnant heifers. Five heifers were fed an iodine deficient diet (mean [sd] iodine concentration 0.06 [0.01] mg/kg dry matter [DM]) and six received an iodine sufficient diet (mean [sd] iodine concentration 1.45 [0.27] mg/kg DM). The diets consisted of wheat and soyabean meal with added minerals and vitamins (with or without iodine) and were fed to the heifers over the final four to five months of pregnancy. The iodine deficient diet produced clinicopathological changes and pathological changes in the thyroid glands of both the heifers and their offspring. However, all the calves in the iodine deficient group were born clinically normal. PMID- 9413123 TI - Caseous lymphadenitis in a commercial ram stud in Scotland. PMID- 9413124 TI - Influence of barley supplement on plasma concentration of triclabendazole metabolites in sheep. PMID- 9413125 TI - Pathological fracture of distal femora in a young dog. PMID- 9413126 TI - Immunodeficiency syndrome in Irish setters. PMID- 9413127 TI - Bladder calculi in dogs and cats. PMID- 9413128 TI - Copper toxicosis in sheep. PMID- 9413129 TI - Controlling messenger RNA stability in bacteria: strategies for engineering gene expression. AB - Recent advances in the understanding of prokaryotic gene expression have led scientists to look beyond traditional promoter control for new methods of regulating gene expression. A promising, new technique centers on controlling the stability of messenger RNA. To exploit the potential of mRNA stability for gene expression control, it is important to understand the mechanisms of prokaryotic mRNA decay as well as the cellular factors that can be used to enhance bacterial gene expression through mRNA stabilization. Factors involved in controlling prokaryotic mRNA stability such as nucleases, secondary structures, translation influences, and transcription effects are discussed and analyzed within the context of three prevailing mRNA decay theories. Several strategies for manipulating mRNA stability in genetically-engineered cells are developed from these discussions and presented as a future direction in gene expression control. In the near future, it should be possible to use these strategies to control mRNA stability in such applications as pharmaceutical protein production and metabolic pathway design. PMID- 9413130 TI - Study of adenovirus production in serum-free 293SF suspension culture by GFP expression monitoring. AB - The red-shifted S65T mutant green fluorescent protein (GFP) was used to compare the adenovirus (Ad) production and post-infection survival of 293SF and 293S cells in serum-free and serum-containing flask cultures, respectively. The GFP expressing vector permitted the quantification of both the level of GFP expressed by infected cells and the infectious viral content of the cultures by flow cytometry in a simple, fast, sensitive, and reliable way. The GFP has the main advantage of fluorescing without any substrate addition. Infected cultures showed the coexistence of two populations of fluorescent cells, high-fluorescence cells (HFCs) and low-flourescence cells (LFCs), in proportions that varied between 20 and 75 hpi. The gradual increase in the number of LFCs at the expense of HFCs correlated well with the increase in the number of dead cells. This relationship could be used for the continuous measure of a culture's viability with the appropriate on-line instrumentation. The post-infection death rate of infected 293SF cells was higher than that of infected 293S cells, but the level of GFP fluorescence in viable, highly fluorescent cells was similar in the two infected cell lines. The number of infectious viral particles (IVPs) was quantified in less than 24 h by an infection assay of 293S cells in wells with viral particles extracted from the culture samples, and the results were more reproducible (+/- 10% variation) than those generally reported for conventional plaque assay titrations or end-point dilutions. The viable cell-specific IVP concentrations were for most experiments similar, indicating again that the difference between the two cell lines was their unequal post-infection viabilities, not the virus production by the infected living cells. PMID- 9413131 TI - Autoregulated multicistronic expression vectors provide one-step cloning of regulated product gene expression in mammalian cells. AB - Regulated expression of a cloned gene often provides much higher final expression of the gene product. Also, regulated expression of an activity can enable optional metabolic engineering and simplify functional genomic research. We constructed di-, tri-, and quattrocistronic mammalian expression vectors which allow the simultaneous, coordinated, and adjustable expression of up to two product genes. A single, tetracycline-regulatable promoter, PhCMV*-1, drives high level expression of a multicistronic expression unit, containing the product gene(s), the gene for tetracycline-responsive transactivator (tTA), and, in the case of pQuattro-tTA, also the neomycin resistance gene. This autoregulatory genetic configuration retains a very low basal transcription activity in the presence of tetracycline, thereby reducing or eliminating possible toxic effects of tTA expression. However, upon withdrawal of tetracycline, a positive feedback regulation loop is activated which leads to higher levels of tTA expression and consequently also to higher expression levels of all other cistrons encoded on the multicistronic expression unit. Since such multicistronic expression vectors combine all genetic elements necessary for high-level expression as well as regulation in a single multicistronic expression unit, they alleviate limitations of previously reported tetracycline-regulatable vector systems and allow straightforward, one-step genetic engineering of eucaryotic cells to give an adjustable phenotype under strict control of the external stimulus, here tetracycline. Because the expression vectors described here were used for the expression for several heterologous product genes such as the green fluorescent protein and the tumor suppressor gene p21 in several cell lines (CHO-K1, BHK-21, and HeLa), we expect these multicistronic, positive feedback regulation vectors to function in a wide variety of eucaryotic cells and to be useful for basic as well as for applied research applications. Other vectors based upon the same autoregulation and multicistronic expression concepts can be constructed using other regulator gene-regulated promoter elements. PMID- 9413132 TI - Application of solution equilibrium analysis to in vitro RNA transcription. AB - Solution equilibrium analysis of in vitro RNA transcription has been applied to examine changes in pH, free magnesium concentration, and concentrations of all chemical ionization species as a transcription reaction proceeds. With this method, the progress of a transcription reaction can be accurately determined as a function of measured pH. In addition, it is demonstrated that this method has significant value as a tool for achieving improved understanding of the effects of varying solution conditions on the dynamics of RNA transcription. Magnesium concentration was found to be a critical factor for efficient transcription. Below 5 mM free Mg2+ concentration, the transcription rate and the efficiency at which nucleoside triphosphates (NTPs) are incorporated are greatly reduced. While inorganic pyrophosphate (PPi), a byproduct of the reaction, was found to directly inhibit the rate of transcription, its detrimental effects on transcription were determined to be primarily due to sequestering of magnesium. The PPi forms a precipitate with magnesium which was determined to have a molar composition of 2:1 of Mg:PPi. Transcription rate and efficiency of NTP incorporation are also reduced with increasing ionic strength. It is shown that these reductions can be partially alleviated by replacing chloride with acetate anions. PMID- 9413134 TI - Kinetics of simultaneous saccharification and lactic acid fermentation processes. AB - After pretreatment of corn cob by dilute acid, the lignocellulosic residue was used as raw materials for the simultaneous saccharification and lactic acid fermentation (SSLF). Because of the same optimal temperature and pH requirement as well as the anaerobic condition, the lactic acid fermentation is perfectly compatible with enzymatic hydrolysis of cellulosic materials. In the SSLF processes, the final concentration of lactic acid reached 33.97 g/L with a conversion ratio of 79% based on the consumed cellulose. A mathematical model is suggested to simulate the SSLF process with good agreement. PMID- 9413133 TI - ComA-dependent transcriptional activation of lichenysin A synthetase promoter in Bacillus subtilis cells. AB - ComA is a DNA-binding activator protein which is required for the transcription of several late-growth phase expressed genes including srfA, an operon needed for the development of genetic competence, efficient sporulation, and surfactin production in Bacillus subtilis (B. subtilis). We show here that the ComA protein can also recognize the promoter regulatory region of the lchA, lichenysin A synthetase operon, found in. Bacillus licheniformis (B. licheniformis) when introduced into B. subtilis cells. Mutational analysis of this region suggests that a palindromic sequence upstream of the lchA promoter may be the target for ComA-dependent transcriptional activation. Considering that the comA operon is present in the B. licheniformis chromosome, we propose the similar mechanism of transcriptional activation of the lichenysin A synthetase operon. PMID- 9413135 TI - Metabolic consequences of phosphotransferase (PTS) mutation in a phenylalanine producing recombinant Escherichia coli. AB - E. coli strain PPA305, which has a wild-type PTS system, and PPA316, which utilizes a proton-galactose symport system for glucose uptake, were used as host strains to harbor a phenylalanine overproduction plasmid pSY130-14 and to study the effects of using different glucose uptake systems on phenylalanine production. The non-PTS strain (PPA316/pSY130-14) produced much less phenylalanine, ranging from 0 to 67% of that produced by the PTS strain (PPA305/pSY130-14) depending on cultivation conditions used. The non-PTS strain PPA316/pSY130-14 had an intracellular PEP concentration only one-sixth that of the PTS strain, PPA305/pSY130-14. Additionally, PPA316/pSY130-14 had a substantially lower energy state in terms of the size of the pool of high-energy phosphate compounds and the magnitude of the pH difference across the cytoplasmic membrane. The non-PTS strain consumed oxygen at a higher rate, attained lower biomass concentration, and produced no acetate and phenylalanine during fermentation, suggesting more carbon was oxidized to CO2, most likely through the TCA cycle. Analysis of intracellular fluxes through the central carbon pathways was performed for each strain utilizing exponential phase data on extracellular components and assuming quasi-steady state for intermediate metabolites. The non PTS strain had a higher flux through pyruvate kinase (PYK) and TCA cycle which, in agreement with the observed higher oxygen uptake rate, suggests that more carbon was oxidized to CO2 through the TCA cycle. Further analysis using rate expression data for PYK and NMR data for the intracellular metabolites identified the regulatory properties of PYK as the probable cause for lower intracellular PEP levels in PPA316/pSY130-14. PMID- 9413136 TI - Reaction engineering for consecutive enzymatic reactions in peptide synthesis: application to the synthesis of a pentapeptide. AB - A single-pot enzymatic synthesis of Z-CCK5 (4-8) is presented in this work, employing Z-Gly-Trp-OBzl as acyl donor, under kinetic control. The first goal of the work is the development of a synthetic strategy allowing the use of the same medium for two reactions catalyzed by immobilized alpha-chymotrypsin, discriminating between simultaneous and consecutive addition systems. The second goal is the maximization of the pentapeptide yield as a function of the molar excess of both nucleophiles employed. A maximum yield of 36% was obtained, and the addition strategy as well as the optimal initial concentrations of substrates have been determined. PMID- 9413138 TI - Kinetics of growth and ribosome-inactivating protein production from Trichosanthes kirilowii plant cell cultures in a 5-L bioreactor. AB - Ribosome-inactivating proteins, named for their ability to inhibit protein translation in cell-free systems, are an important class of natural plant defense proteins with potential human therapeutic and agricultural applications. The kinetics of growth, nutrient consumption, and extracellular protein translation inhibitory activity are presented for Trichosanthes kirilowii plant cell suspensions in 5-L bioreactors at two agitation rates (50 and 100 rpm). The cultures had a 7-9.5 day lag phase followed by exponential growth with a doubling time of less than 2 days. Biomass concentrations reached levels of approximately 19 g (dry weight)/L. Protein translation inhibitory activity was observed in the culture broths during the exponential growth phase and reached levels of approximately 50-60 units. No detrimental effects of agitation were observed at 100 rpm. These studies demonstrate the potential for plant cell culture production of ribosome-inactivating proteins in bioreactor systems. PMID- 9413137 TI - Biotreatment of ammonia from air by an immobilized Arthrobacter oxydans CH8 biofilter. AB - A heterotrophic Arthrobacter oxydans CH8 that was capable of removing NH3 from NH3 containing gas was isolated from livestock farming wastewater. The A. oxydans CH8 was immobilized with calcium alginate packed into filter column. Metered NH3 containing gas was partially humidified and passed through the glass column. Extensive tests including the removal characteristics, the removal efficiencies, and the metabolic products of NH3 by A. oxydans CH8 were conducted. Additionally, the operation criteria for the biofilter was also established. NH3 removel capacities were elevated by the immobilized-cell (biological conversion) method and the BDST (bed depth service time) method (physical adsorption), respectively. The optium temperature for removing NH3 was 30 degrees C, while the nitrification ability remained 80% at 40 degrees C. The high efficiency (> 97%) in the removal of NH3 was attained at 36 L/h with pH control and was not decreased because of high NH3 inlet concentration. In addition, the high maximum removal rate (1.22 g of N/day (kg of bead)) enhanced the use of the biofilter in industrial-scale NH3(g) pollution control. The ability to remove NH3 at high inlet concentration and temperature suggested that the immobilized A. oxydans CH8 biofilter has potential in processing NH3 gas. PMID- 9413139 TI - Enzymatic large-scale production of 2-keto-3-deoxy-D-glycero-D-galacto nonopyranulosonic acid in enzyme membrane reactors. AB - The enzymatic synthesis of 2-keto-3-deoxy-D-glycero-D-galacto-nonopyranulosonic acid (KDN) starting from D-mannose and pyruvic acid using Neu5Ac-aldolase has been scaled up. A repetitive batch ultrafiltration bioreactor was used for the KDN synthesis on 100 g scale with a conversion of up to 85%. Furthermore, a 440 mL pilot-scale enzyme membrane reactor (EMR) was performed for the continuous production of KDN. Conversion of mannose was 75% at a space--time yield of 375 g/(L d). KDN was advanteageously isolated by crystallization with an overall yield of 75%. PMID- 9413140 TI - Microbial removal of alkanes from dilute gaseous waste streams: kinetics and mass transfer considerations. AB - Treatment of dilute gaseous hydrocarbon waste streams remains a current need for many industries, particularly as increasingly stringent environmental regulations and oversight force emission reduction. Biofiltration systems hold promise for providing low-cost alternatives to more traditional, energy-intensive treatment methods such as incineration and adsorption. Elucidation of engineering principles governing the behavior of such systems, including mass transfer limitations, will broaden their applicability. Our processes exploit a microbial consortium to treat a mixture of 0.5% n-pentane and 0.5% isobutane in air. Since hydrocarbon gases are sparingly soluble in water, good mixing and high surface area between the gas and liquid phases are essential for biodegradation to be effective. One liquid-continuous columnar bioreactor was operated for more than 30 months with continued degradation of n-pentane and isobutane as sole carbon and energy sources. The maximum degradation rate observed in this gas-recycle system was 2 g of volatile organic compounds (VOC)/(m3.h). A trickle-bed bioreactor was operated continuously for over 24 months to provide a higher surface area (using a structured packing) with increased rates. Degradation rates consistently achieved were approximately 50 g of VOC/(m3.h) via single pass in this gas-continuous columnar system. Effective mass transfer coefficients comparable to literature values were also measured for this reactor; these values were substantially higher than those found in the gas-recycle reactor. Control of biomass levels was implemented by limiting the level of available nitrogen in the recirculating aqueous media, enabling long-term stability of reactor performance. PMID- 9413141 TI - Thermal unfolding of bacteriophage T4 short tail fibers. AB - The short tail fibers of bacteriophage T4 are composed of a homotrimer of the product of gene 12 (P12) with a molecular weight of 165,000. P12 is capable of reconstituting defective phage particles lacking gene 12. After heating to 75 degrees C, P12 was able to retain 90% of its ability to reconstitute T412- particles. When heated above 75 degrees C, P12 was no longer capable of fully reconstituting defective phage particles. By 95 degrees C, the reconstitution efficiency of the P12 preparation was reduced by 4 orders of magnitude. Thermal unfolding was also monitored by heating the protein in the presence of SDS to freeze partially unfolded states; by protease hydrolysis; and by intrinsic fluorescence changes. Exposure to SDS had little effect for temperatures up to 55 degrees C, but by 65 degrees C, the reconstitution efficiency of P12 treated with 0.01% SDS dropped to less than 1% of the original titer. Thermolysin digestion of P12 heated to various temperatures showed that treated P12 started to inactivate before 45 degrees C and inactivation was essentially complete by 55 degrees C. Intrinsic fluorescence data of heated P12 indicated that the protein begins to unfold by 45 degrees C and exhibits distinct peak shifts at 60 degrees C and above 80 degrees C. We conclude that, in the absence of SDS or proteases, P12 heated to 75 degrees C can refold back to an active conformation. Trimeric P12 undergoes some irreversible denaturation between 75 and 85 degrees C, and heating between 85 and 95 degrees C results in dissociation of P12 into monomers. PMID- 9413142 TI - Immunomagnetic isolation of islets from the rat pancreas. AB - Islets were selectively isolated from rat pancreatic digests using magnetic microspheres coated with anti-islet monoclonal antibodies. The isolation process was optimized as a function of bead concentration and time of incubation with the tissue digest. Apparent and normalized islet yields of 92 +/- 6% and 81 +/- 7%, respectively, were obtained by incubating the digests with 10(7) beads/mL for 40 min followed by isolation of the bead-coated islets under a magnetic field. While some fragmentation occurred, the isolation process did not alter islet functionality as demonstrated by an insulin secretion response to glucose stimulation equivalent to that of handpicked controls. The technique is fast, reproducible, and potentially scaleable to larger pancreases as a secondary purification step. PMID- 9413143 TI - Effect of ethanol, ammonium sulfate, fatty acids, and temperature on the solution behavior of bovine serum albumin. AB - Ternary phase diagrams (TPDs) for aqueous bovine serum albumin (BSA) solutions containing ethanol or (NH4)2SO4 were determined for temperatures ranging from 20 to 70 degrees C. At 20 degrees C, ethanol destabilized BSA, resulting in gel formation at moderate solute concentrations. In contrast, (NH4)2SO4 concentrations above 20 wt % precipitated BSA from solution. Raising the temperature led to gel formation at increasingly lower BSA and solute concentrations. The removal of adsorbed fatty acids from BSA had little effect on the ethanol TPDs but reduced protein solubility in (NH4)2SO4. Moreover, the salt TPDs of fatty-acid-poor BSA were similar to those previously observed with S ovalbumin. PMID- 9413144 TI - Glassy state and thermal inactivation of invertase and lactase in dried amorphous matrices. AB - The thermal stability of enzymes lactase and invertase in dried, amorphous matrices of sugars (trehalose, maltose, lactose, sucrose, raffinose) and some other selected systems (casein, PVP, milk) was studied. The glass transition temperature (Tg) was limited as a threshold parameter for predicting enzyme inactivation because (a) enzyme inactivation was observed in glassy matrices, (b) a specific effect of enzyme stabilization by certain matrices particularly trehalose was observed, and (c) enzyme stability appeared to depend on heating temperature (T) "per se" rather than (T-Tg). For these reasons, a protective mechanism by sugars related to the maintenance of the tertiary structure of the enzyme was favored. A rapid loss of enzyme (lactase) activity was observed in heated sucrose systems at T > Tg, and this was attributed to sucrose crystallization since it is known that upon crystallization the protective effect of sugars is lost. Thus, the stabilizing effect could be indirectly affected by the Tg of the matrix, since crystallization of sugars only occurs above Tg. Trehalose model systems (with added invertase) showed an exceptional stability toward "darkening" (e.g., non-enzymatic browning) when heated in the dried state to elevated temperatures and for long periods of time. PMID- 9413145 TI - Insertion of stabilizing loci in vectors of T7 RNA polymerase-mediated Escherichia coli expression systems: a case study on the plasmids involving foreign phospholipase D gene. AB - Plasmids carrying stabilizing loci were used in the expression of phospholipase D (PLD) gene fused with pelB signal sequence by a recombinant strain of Escherichia coli BL21 (DE3) using T7 RNA polymerase mediated expression system. By checking the living cell number and the percentage of the plasmid-bearing cells, it was found that the plasmids involving PLD gene were not stable under noninduced conditions and that, after the induction, the number of plasmid-bearing cells were rapidly decreased to almost zero. Then, a biologically stabilizing locus such as par B, ccd, or par was inserted into the plasmids. The newly constructed plasmids were maintained very stably in the recombinant cells until the cells were induced. However, after the induction, almost all the recombinant cells were rapidly killed due to highly toxic PLD. Using the best one of the stabilized PLD expressing plasmids, PLD production was improved 2-fold. PMID- 9413146 TI - Bioassay using a labeled oligonucleotide obtained by in vitro selection. AB - A new bioassay system using an oligodeoxyribonucleotide (DNA) obtained by the in vitro selection method is described. The DNA aptamer which selectively binds to a target molecule, Reactive Green 19 (RG19), was labeled with fluorescein. The circular dichroism spectra revealed that the fluorescein labeling did not significantly affect the conformation of the DNA. The binding affinity toward RG19 of the labeled DNA was the same as that of the nonlabeled DNA. Using the labeled DNA and the RG19-immobilized gel, a semiquantitative assay of RG19 concentration was performed. PMID- 9413147 TI - Altered efflux properties of mouse leukemia L1210 cells resistant to 4-methyl-5 amino-1-formylisoquinoline thiosemicarbazone. AB - A mouse leukemia L1210 cell line, denoted MQ-580, that was selected for resistance to the ribonucleotide reductase inhibitor, 4-methyl-5-amino-1 formylisoquinoline thiosemicarbazone (MAIQ), in addition to having altered properties at the ribonucleotide reductase site had other alterations that contributed to its resistant phenotype; these included the expression of p glycoprotein and the multi-drug resistance associated protein (MRP). The efflux of rhodamine 123 (Rh-123) or daunomycin (Dau) was greatly increased in MQ-580 cells compared to parental wild-type (WT) cells. The effluxes of Rh-123 and Dau were ATP- and temperature-dependent. The p-glycoprotein inhibitors, verapamil, cyclosporin A and reserpine blocked the efflux of both Rh-123 and Dau. In contrast, the inhibitors of MRP, MK571, BSO-treatment, arsenite and genistein did not block the efflux of either Rh-123 or Dau from MQ-580 cells. These findings suggest that the p-glycoprotein is the major transporter involved in effluxing Rh 123 and Dau from MQ-580 cells. PMID- 9413148 TI - Triphenylselenonium and diphenylselenide in cancer chemoprevention: comparative studies of anticarcinogenic efficacy, tissue selenium levels and excretion profile. AB - The objectives of the present study were to evaluate the cancer chemopreventive activity of triphenylselenonium chloride and diphenylselenide and to investigate the pharmacology of these two compounds with respect to their tissue accumulation and excretion profile. Although both phenyl selenide derivatives are related to each other structurally, they differ substantially in their intrinsic chemical properties. Triphenylselenonium is positively charged and amphiphilic, while diphenylselenide is uncharged and lipophilic. With the use of either the DMBA- or MNU-induced mammary tumor model in rats, triphenylselenonium was found to have superior chemopreventive efficacy compared to diphenylselenide. Both reagents were present at 30 ppm Se in the diet. At the time of sacrifice (22 weeks post carcinogen), triphenylselenonium produced only minimal accumulation of selenium in the liver, kidney, mammary gland and plasma. In contrast, diphenylselenide caused a 2- to 3-fold elevation in selenium concentration depending on the tissue examined. Thus even though diphenylselenide was able to increase total selenium in tissues, it was less active in cancer protection. Fecal excretion following a single oral dose of triphenylselenonium (equal to the amount consumed in 1 day by an animal fed a diet containing 30 ppm Se) was approximately 78% and 8% of the dose during the first and second day, respectively, suggesting that the bulk of the dose was not absorbed. With diphenylselenide, fecal excretion was about 6% and 30% of the dose during the first and second day, and about 20% of the dose was excreted in the urine in each of the 2 days. This observation suggests that a large proportion of the diphenylselenide dose was absorbed and that urinary excretion was a major route of elimination for diphenylselenide once it was absorbed. Further studies are needed to clarify the basis for the differential effects of these phenyl selenide derivatives. PMID- 9413149 TI - Tumor cell invasion through matrigel is regulated by activated matrix metalloproteinase-2. AB - We tested the hypothesis that there is a correlation between tumor cell efficiency in activation of matrix metalloproteinase-2 (MMP-2) and invasion through basement membrane-like Matrigel barriers. To generate cells capable of MMP-2 activation, we stably transfected three human tumor cell lines, HT-1080 fibrosarcoma, MCF7 breast carcinoma, and U251.3 glioma with cDNA encoding the full length human membrane-type matrix metalloproteinase-1. Our results show a bimodal correlation between the extent of MMP-2 activation and Matrigel invasion by tumor cells. Cell transfectants characterized by a partial activation of MMP-2 were the most invasive while those with an extensive conversion of MMP-2 proenzyme into enzymatically active forms were the least efficient in invading Matrigel. Modulation of MMP-2 activation by exogenous TIMP-2 reverted the rate of Matrigel invasion by cell transfectants to control levels. We conclude that the regulation of activated MMP-2 in the tumor cells, microenvironment may be critical in facilitating tumor cell invasiveness. PMID- 9413151 TI - Interleukin 10 is expressed in human gliomas in vivo and increases glioma cell proliferation and motility in vitro. AB - Interleukin 10 (IL-10) is a cytokine with a broad spectrum of immunosuppressive activity, but itoffs role in the oncogenesis of solid tumors is still unclear. In previous experiments we have shown that IL-10 specific mRNA is produced within glial tumors in vivo. The aim of the present study was to investigate the expression of the IL-10 protein in vivo and to identify the cells producing IL-10 within the tumor tissue. Expression levels significantly increased with malignancy of the gliomas. 87.5% of grade III and IV, but only 4% of grade II tumors expressed high levels of mRNA. Elevation of IL-10 serum levels was found in 11% of low grade and in 63.6% of high grade glioma patients. In situ hybridization analysis with combined immunohistochemistry revealed that: a) IL-10 is not produced by infiltrating B- or T- lymphocytes, b) both microglia and astroglia contributed to IL-10 expression in malignant gliomas in vivo. These data suggested the functional role of IL-10 in glioma progression. Therefore, the effects of IL-10 on proliferation and migration of glioma cells were determined in vitro. Two human glioma cell lines were grown as monolayer as well as spheroids in the presence of different concentrations of IL-10. IL-10 increased cell proliferation significantly in both culture systems with a dose optimum of 25 ng/ml. Glioma cell motility was enhanced with 25 ng/ml as the optimal dose. Adding the IL-10 specific antibody reversed both effects. We conclude from our data that IL-10 is involved in the progression of glial tumors, especially in the enhancement of tumor cell proliferation and migration which promotes infiltration of the surrounding tissue. PMID- 9413150 TI - Transcriptional activation of the bcl-2 apoptosis suppressor gene by the paired box transcription factor PAX8. AB - The bcl-2 proto-oncogene suppresses apoptosis in a variety of cell types and is essential for normal renal development. PAX8 is a member of the paired box class of transcription factors and is developmentally regulated. bcl-2 and PAX8 are both expressed in the kidney during development and throughout life, demonstrating a nearly identical pattern of expression. We used transient transfection reporter assays and electrophoretic mobility shift assays to test PAX8 transcriptional regulation of bcl-2. PAX8 transcriptionally activates the bcl-2 promoter and binds to promoter sequences in vitro. These findings establish that PAX8 can transcriptionally regulate bcl-2 and suggest that suppression of apoptosis is an important event in the development and maintenance of renal tubular epithelial structures. PMID- 9413152 TI - Oncogene and HSP-70 expression in primary tumor cell cultures of renal cell carcinoma compared to their corresponding cell line. AB - BACKGROUND: Tumor progression in renal cell carcinoma (RCC) can be explained by a multistep model, in which the activation of certain oncogenes such as c-neu and c fos appear to be early events in tumorigenesis, while the expression of p53 and pan-ras are found in advanced stages. MATERIAL AND METHODS: The expression of oncogenes and growth factors was examined in 29 primary tumor cell cultures (PTCC) of RCC using immunocytochemistry. RESULTS: In PTCC high expression of c neu and c-fos was present in all tumors, whereas mdr, TGF-alpha, EGF, c-myc, pan ras, p53 and HSP-70 was detected at low expression levels. In 27% (8/29) of PTCC, cell lines (CL) were established. Oncogene expression was increased in CL compared to PTCC. CONCLUSION: The pattern of oncogene expressions found in CL is similar to findings described in highly malignant tumors in vivo. Therefore, the establishment of CL seems to depend on a selective recruitment of tumor cells with an upregulated oncogene expression. PMID- 9413153 TI - Detection of neutral endopeptidase 24.11 (neprilysin) in human hepatocellular carcinomas by immunocytochemistry. AB - BACKGROUND: We have reported previously that neutral endopeptidase 24.11 (neprilysin; NEP; CALLA, CD10) activity was very high in rat hepatomas and a cultured human hepatocarcinoma cell line (SK-HEP1). MATERIALS AND METHODS: While continuing these studies, we detected the presence of NEP in SK-HEP 1 cells by immunocytochemistry and in paraffin-embedded human hepatocellular carcinomas as well. IgG purified from polyclonal antisera to human NEP was employed as a source of antibody. RESULTS: SK-HEP 1 cells gave a strong positive reaction to the IgG fraction of the antisera. In control studies, where IgG was preabsorbed with recombinant NEP, the results were negative. Of the 18 hepatocellular carcinomas tested, NEP was expressed in 14 (78%) malignant tumors, while adjacent liver tissue did not show the presence of NEP. CONCLUSIONS: It is suggested that, because none of the known hepatocellular carcinoma markers are highly specific, the detection of NEP in these malignant cells can be an additional useful diagnostic tool. PMID- 9413154 TI - Antimetastatic activity of the new platinum analog [Pt(cis-dach) (DPPE).2NO3] in a metastatic model of human bladder cancer. AB - Surgical orthotopic implantation (SOI) of histologically intact human RT-4 bladder tumor tissue in nude mice resulted in local growth, invasion, regional extension and metastases as well as distant metastases to other organ sites and lymph nodes, thus mimicking the bladder cancer patient. This metastatic bladder tumor animal model was treated with two doses of new platinum analog ?Pt(cis dach)(DPPE).2NO3? for the evaluation of antimetastatic efficacy compared to two doses of cisplatinum. Unlike the untreated control group or the group treated with the low dose of cisplatinum, there were no metastases in either the high or low-dose platinum-analog-treated groups and the high-dose cisplatinum-treated group. The results obtained with this patient-like nude-mouse model of bladder cancer indicate that the new platinum analog appears to be a valuable lead compound with antimetastatic efficacy and clinical potential. PMID- 9413155 TI - Growth inhibition of a human lung adenocarcinoma cell line by genetic complementation with chromosome 11. AB - Two regions on chromosome segment 11p15.5 have frequent allele loss in lung cancer. LOH11A is centromeric between loci D11S1758 and D11S12, and LOH11B is telomeric between HRAS and D11S1363. We studied the biological significance of this allele loss using microcell-mediated transfer of human chromosomes 11, 11p, and two radiation-reduced fragments of 11p into human lung adenocarcinoma cell lines. Chromosome 12, which has not been implicated in lung carcinogenesis, was used as a control. All four chromosome 11-containing hybrid clones showed significantly reduced tumorigenicity in nude mice and growth in liquid culture. These findings support the notion of a tumor suppressor gene located in the LOH11A region on chromosome segment 11p15.5. PMID- 9413156 TI - Interleukin 18 enhances Fas ligand expression and induces apoptosis in Fas expressing human myelomonocytic KG-1 cells. AB - Interleukin-18 (IL-18) induces apoptosis in human myelomonocytic KG-1 cells as determined by agarose gel electrophoresis, and flow cytometry after propidium iodide (PI) staining. Apoptosis was detected 20 hours from the start of culture at concentrations of 100 ng/ml of the cytokine. Although IL-18 induces the production of large amounts of interferon gamma (IFN-gamma) by KG-1 cells, conditioned media could not induce apoptosis of fresh cells. The protein expressions of p53 and Fas ligand by KG-1 cells, which constitutively express the Fas antigen (CD95), were found to increase after exposure to IL-18 for 20 hours. Both Fas ligand and its receptor were found to be functional by in vitro assays on Fas-expressing target cells and an agonist anti-Fas antibody, respectively. In conclusion, IL-18 enhances the expression of Fas ligand by Fas-expressing KG-1 cells and induces apoptosis in the cells through a mechanism probably involving the Fas pathway. PMID- 9413157 TI - Estradiol induces DNA fragmentation in a human endometrial adenocarcinoma with estradiol-inhibited growth phenotype. AB - A moderately differentiated human endometrial adenocarcinoma heterotransplanted into nude mice was investigated for morphological and molecular changes in the tumours after treating the animals with estradiol. The tumour growth was previously characterised as estradiol-independent but responsive (inhibited) without any changes in cell proliferation. In response to hormonal treatment rather the cell loss factor increased. In this experiment tumours influenced by estradiol were investigated at different time-points after treatment by an in situ labelling technique to detect cells undergoing DNA fragmentation as a sign of apoptosis. Expression of the apoptosis related protein bcl-2 was evaluated by Western blotting. Tumours from animals treated with estradiol showed an increase in tumour volume doubling time from 5.4 days to 16 days compared to control tumours. Histologically, tumours influenced by estradiol were better differentiated than control tumours and showed a significant increase in cells staining positively with the in situ apoptosis detection technique. A parallel time dependent decreased expression of bcl-2 protein was observed. These results confirm our previous findings where estradiol influenced the cell loss factor without changes in the growth fraction, indicating increased apoptotic activity in response to hormonal treatment. PMID- 9413158 TI - Interaction of the Rb tumor suppressor protein with the c-fos promoter in c-fos transfected cells overexpressing c-fos and Rb. AB - The Rb tumor suppressor protein is overexpressed in HeLa cell lines permanently transfected with a constitutively expressed c-fos gene. CP17-14 cell overexpression of Rb may be due to a balancing response to overexpression of the stimulatory effects of c-fos overexpression on transcription. The cis-acting retinoblastoma control element (RCE, -97 to -86 bp) in the human c-fos promoter is thought to allow regulation of c-fos by Rb. Gel-shift assays were performed with a 168 bp fragment encoding the c-fos RCE. Competition assays with increasing mass of unlabeled probe or dose-dependence assays using increasing mass of nuclear proteins, demonstrated sequence-specific complex formation. Indistinguishable complexes were formed between the c-fos RCE fragment in transfected cells, but at higher levels (> 50%), compared to proteins from parental cells. Supershift analysis utilizing epitope-specific Rb-monoclonal antibodies indicated the presence of Rb protein bound to the RCE-containing DNA fragment. In contrast, polyclonal anti-Rb antibodies enhanced the amounts of nuclear protein-DNA complexes detected but did not result in a supershift. These results suggested the presence of Rb and/or Rb-like peptides involved in complex formation and the presence of multiple variants of RCE-binding complexes in response to c-fos over-expression. PMID- 9413159 TI - Inhibition of pp60c-src protein kinase by herbimycin A in polyomavirus middle tumor antigen-transformed cells. AB - The middle T antigen (MTAg) encoded by polyomavirus plays an important role in virus-mediated tumorigenesis. The activated protein kinase activity of MTAg associated pp60c-src has been shown to be necessary for cell transformation by polyomavirus. In this study, the effects of herbimycin A on the pp60c-src kinase activities in the polyomavirus- and MTAg-transformed cells were studied. Phosphorylation of src and MTAg is reduced in polyomavirus and MTAg- transformed cells pretreated with herbimycin A. Inactivation of the enzymatic activity by herbimycin A was found to be dependent on the time of incubation and the drug concentration. In contrast, src immunoprecipitates from untreated MTAg transformed cells appeared to be resistant to inhibition by herbimycin A. Herbimycin A does not affect the synthesis of MTAg and pp60c-src in the MTAg transformed cells. These results suggest that pp60c-src kinase activity in the drug-treated cell lysates is more sensitive to herbimycin A inhibition than the same activity in the immunoprecipitates. PMID- 9413160 TI - 5q-: does longer survival of female patients explain the preponderance. AB - Cases with deletion of part of the long arm of chromosome 5 (5q-) and a myeloid hemopoietic disorder show a marked female preponderance. We analysed the female preponderance of 328 published cases of myelodysplastic syndromes (MDS) or acute myeloid leukemia (AML). MDS without clinical or cytogenetic progression shows by far the highest frequency in female patients. Cases with advanced MDS and/or chromosome aberrations in addition to 5q- show low to moderate preponderance. Female preponderance was found to correlate positively with relative survival: in the group with the highest frequency women survived significantly longer than men. In groups with lower preponderances the survival of the two sexes gradually converged. These data suggest that in 5q- related disorders the female preponderance is a consequence of increased female survival. PMID- 9413161 TI - Effect of dibutyryl cyclic AMP on the cyclin-dependent kinase inhibitor p27Kip1 in the human hepatoma cells PLC/PRF/5. AB - The cyclin-dependent kinase (cdk) inhibitor p27Kip1 is known to play a role in cell-cycle regulation at G1 and G1/S phase. We investigated the effect of the putative growth-inhibiting agent dibutyryl cyclic AMP (DBcAMP) on the serial changes of p27Kip1 expression in the human hepatoma cells PLC/PRF/5 in culture. The p27Kip1 protein level increased at an early stage of G1 phase (2 hours) after a release from serum-starvation and subsequently maintained the level until the entry to S phase, whereas an addition of DBcAMP at 1mM increased the p27Kip1 protein level during G1 phase. In contrast, the relative expression levels of p27Kip1 mRNA at 2 hours, 4 hours and 6 hours were lower in DBcAMP-added cells. The effects of DBcAMP on cell growth were, reduction of S-phase cells, inhibition of DNA synthesis, and accumulation of G2-phase cells. In the presence of the antisense oligodeoxynucleotides against p27Kip1 mRNA, DBcAMP-induced growth inhibition was partially abolished. These findings suggest that DBcAMP elevates p27Kip1 protein expression during G1 phase, which could be associated with growth inhibition. DBcAMP may inhibit the degradation of p27Kip1 protein. PMID- 9413162 TI - Inhibition of xanthine oxidase by hydroxylated anthraquinones and related compounds. AB - Eighteen anthraquinones and related compounds were tested for their inhibitory effects on xanthine oxidase. The enzyme, xanthine oxidase catalyses the oxidation of hypoxanthine to xanthine and of xanthine to uric acid, which has a lambda max of 295nm, forming the basis for a spectrophotometric assay of the activity of xanthine oxidase. The results showed that anthrarobin and purpurin showed moderate effects on xanthine oxidase inhibition (IC50 = 68.35 and 105.13 microM; Ki = 122.38 and 130.49 microM respectively), and both of them induced mixed type (competitive-non-competitive) inhibition with respect to the substrate xanthine. PMID- 9413163 TI - Synthesis and antitumor activity of boronated dipeptides containing aromatic amino acids. AB - N-[(Trimethylamine-boryl-carbonyl]-L-tryptophan methyl ester and N[(trimethylamine-boryl)-carbonyl]-L-histidine methyl ester were obtained by synthesis using triphenyl-phosphine/carbon tetrachloride or dicyclohexyl carbodiimide as coupling agents, respectively. Both agents reduced L1210 lymphoid leukemia DNA, RNA, and protein syntheses with the largest reductions occurring in DNA synthesis. Reductions in DNA synthesis appear to be mediated by inhibition of key enzyme activities (i.e., DNA polymerase a, IMP dehydrogenase, and PRPP amido transferase). These agents had little effect on in vitro L1210 DNA topoisomerase II activity at 100 microM but were able to cause synergistic increases in protein linked DNA breaks when combined with etoposide (VP16). It was shown that these agents significantly reduced protein kinase C mediated phosphorylation of human topoisomerase II in vitro. Thus, inhibition of topoisomerase II phosphorylation may be a mechanism by which these agents and VP-16 are synergistic in causing protein-linked DNA breaks. PMID- 9413164 TI - Development of a hammerhead ribozyme against BCL-2. II. Ribozyme treatment sensitizes hormone-resistant prostate cancer cells to apoptotic agents. AB - BACKGROUND: Several lines of evidence strongly implicate a crucial role for the apoptosis suppressing bcl-2 oncogene in the genesis of hormone-refractory human prostate cancer. By efficiently destroying the intracellular bcl-2 mRNA, one might be able to make the prostate cancer cell responsive again to conventional apoptotic stimuli such as androgen withdrawal. To achieve this end, we have devised a catalytic antisense RNA strategy (Ribozyme) for bcl-2 and evaluated its gene therapeutic potential. METHODS AND RESULTS: Bcl-2 overexpressing LNCaP prostatic carcinoma cells (LNCaP/bcl-2) were transfected with the anti-bcl-2 ribozyme RNA using a polyamine-based transfection reagent and the reduction in the intracellular bcl-2 mRNA levels was followed by a ribonuclease protection assay. Using a cell viability assay, prior ribozyme transfection and subsequent application of apoptotic stimuli such as serum starvation or phorbol ester treatment caused a 30% increase in cell death by apoptosis than with these apoptotic stimuli alone. CONCLUSIONS: The results obtained strongly support the ability of a potential anti-bcl-2 ribozyme therapy to synergize with other agents in inducing apoptosis of hormone-resistant human prostate cancer cells. PMID- 9413165 TI - The new sensitizing agents for photodynamic therapy: 21-selenaporphyrin and 21 thiaporphyrin. AB - BACKGROUND: Photodynamic therapy may be a promising treatment for patients with tumors. The mechanism of its action is poorly understood and different from the cytotoxic effects induced by antitumor drugs. MATERIALS AND METHODS: New sensitizers, termed as 21-selenaporphyrin (SEP) and 21-thiaporphyrin (STSP) were studied for their photocytotoxicity in vitro against selected human cancer cell lines. This study was followed by in vivo screening of the effect of SEP using an animal tumor model. The activity of the new agents was compared with that of a known photosensitizer, namely chlorin e6. In our selection of the cell lines applied for in vitro study, the possible accessibility and effectiveness of photodynamic therapy (PDT) for treatment of colon and urinary bladder cancers, was considered. RESULTS: New compounds appeared to be not toxic for tested cells in culture, without exposure to light. The STSP exerted in vitro effects comparable with chlorin e6 photocytotoxicity, while SEP appeared to be ineffective. However, in vivo experiments performed in a BFS1 fibrosarcoma tumor model in mice showed that the SEP was at least as much effective as chlorin e6 in the induction of tumor necrosis. In contrast to chlorin e6, SEP-PDT induced no skin sensitization. CONCLUSIONS: Both new sensitizers can be applied in PDT at no risk of skin damage. The mechanism of the action of these two compounds is probably different, i.e. the 21-thiaporphyrin possibly acts directly on tumor cells and the 21-selenaporphyrin via endothelial cells of newly formed tumor vasculature. PMID- 9413166 TI - Drug resistance conferred by MDR1 expression in spheroids formed by glioblastoma cell lines. AB - Three-dimensional cell cultures (spheroids) provide an in vitro approximation of solid tumors in vivo. Some mechanisms of drug resistance were reported to differ in their effects on monolayer and spheroid cells. We have compared the effect on the expression of P-glycoprotein (Pgp), the MDR1 gene product, on vinblastine resistance in two glioblastoma lines, U87 and U251, grown as either monolayers or spheroids. U87 cells form spheroids spontaneously and the spheroids continue to grow, while U251 cells form spheroids only when plated over agarose and do not proliferate in spheroids. U87 cells are equally resistant to vinblastine in monolayer and in spheroids, while U251 cells are 4.5 times more resistant in spheroids than in monolayers. The distribution of a fluorescent vinblastine derivative in cells of spheroids was more heterogeneous than in monolayers. MDR1 expressing U87 cells increased their resistance 9-fold in monolayer and 50-fold in spheroids, while MDR1 transduction of U251 cells made them 20-fold more resistant in monolayer and 160-fold more resistant in spheroids. MDR1 expression in both cell lines decreased the accumulation of 3H-vinblastine and fluorescent vinblastine derivative. These results indicate that MDR1 is more efficient in conferring drug resistance in spheroids than in monolayer cells. PMID- 9413167 TI - Pulsed exposure of SDZ PSC 833 to multidrug resistant P388/ADR and MCF7/ADR cells in the absence of anticancer drugs can fully restore sensitivity to doxorubicin. AB - AIM: An in vitro cellular pharmacological study was undertaken to characterize the latency of modulating activity by the chemosensitizer, PSC 833. METHODS: PSC 833 was evaluated for cytotoxicity in the MDR P388/ADR and MCF7/ADR cell lines. Cellular uptake levels and intracellular DOX distribution characteristics were assessed by flow cytometry and fluorescence microscopy, respectively. These parameters were correlated with the chemosensitivity of the MDR cells under varying conditions of exposure to PSC 833 and DOX. RESULTS: PSC 833 (1 microM) provided near complete MDR reversal and exhibited significant latent modulating activity indicative of a sustained inhibition of the PGP pump in P388/ADR and MCF7/ADR cells after removal of the MDR modulator. Interestingly, complete latent chemosensitizing activity could be achieved by pulsing cells with PSC 833 prior to DOX exposure. Increases in cellular DOX uptake by individual P388/ADR and MCF7/ADR cells induced by MDR modulators correlated well with their ability to chemosensitize the resistant tumor cells where intracellular DOX levels approaching those obtained for sensitive wild type cells was observed for PSC 833. CONCLUSION: The MDR modulator PSC 833 exhibits latent MDR modulating activity that appears well suited for modulating PGP mediated MDR in vivo where chemosensitization is complicated by difficulties in synchronizing therapeutic levels of modulator and anticancer drug at the tumor site. PMID- 9413168 TI - Chemotherapeutic evaluation of N-(4-hydroxyphenyl) retinamide-O-glucuronide in the rat mammary tumor model. AB - The growth inhibitory effects of N-(4- Hydroxyphenyl) retinamide (4-HPR) and its glucuronide derivative, N-(4-Hydroxyphenyl) retinamide-O-glucuronide (4-HPROG) on established DMBA induced rat mammary tumors were compared. The results indicate that the glucuronide analog had a greater antitumor potency than equimolar concentration of the free retinoid. Tumor regression occurred in 75% of the rats fed 2 mmol/Kg diet of 4-HPROG. In a 6-week study, the maximum tolerated dietary dose (MTD) was found to be 3.5 mmol/Kg diet for 4-HPR and 5 mmol/Kg diet in the case of 4-HPROG. The higher potency and lower toxicity of the glucuronide suggests that this conjugate may have an in vivo chemotherapeutic advantage over the parent free retinoid. PMID- 9413169 TI - Evidence that DNA methylation imbalance is not involved in the development of malignant mesothelioma. AB - Methylation dysregulation has been a consistent finding in various malignancies, particularly those where the pathogenetic mechanisms are unclear. In order to test the hypothesis that methylation imbalance may not be a feature of cancers where the aetiologic agent or process is known, we studied the methylation status of the myogenic genes Myf-3 and Myf-4 by Southern blotting in malignant mesothelioma, a cancer strongly associated with asbestos exposure. DNA samples obtained from controls and mesothelioma patients did not exhibit hypermethylation of Myf-3 and hypomethylation of Myf-4, as noted in malignant lymphomas. The methylation status of Myf-3 and Myf-4 in malignant mesothelioma was similar to that of non-malignant cells indicating that dysregulation of the DNA methylating machinery may not be involved in mesothelioma development. The present findings do not support the view that methylation imbalance is a consequence of neoplastic transformation, but indicate that it may be one of the early molecular events involved in the genesis of some cancers. PMID- 9413170 TI - Exposure to sorbitol induces resistance to cisplatin in human non-small-cell lung cancer cell lines. AB - Cisplatin is the most active anticancer agent for lung cancer. It has been reported that intracellular accumulation of cisplatin is important in determining resistance to cisplatin, which may be modulated by Na+, K(+)-ATPase activity. On the other hand, it is well-known that sorbitol, a metabolite of glucose mediated by aldose reductase, reduces Na+, K(+)-ATPase in diabetic neuropathy. In this study, the effect of exogenous sorbitol on Na+, K(+)-ATPase activity and sensitivity to cisplatin was evaluated using human non-small-cell lung cancer (NSCLC) cell lines. In the NSCLC cell lines, EBC-1, PC-3, and RERF-LC-MS the cytotoxicities of cisplatin were impaired by exposure to sorbitol in these cell lines. Na+, K(+)-ATPase was inactivated and intracellular accumulation of cisplatin was decreased by the exposure. These results suggest that accumulation of sorbitol may induce resistance to cisplatin in NSCLC cells, and diabetes poorly controlled may be one of the determinants of the antitumor effect of cisplatin in NSCLC. PMID- 9413171 TI - Inhibition by guanidino compounds of the growth of spontaneous mammary tumours in SHN mice. AB - The D-form of lombricine extracted from the earthworm was previously found to inhibit the growth of spontaneous mammary tumours of mice. In the present study, the anti-mammary tumour effects of guanidino compounds, of which structures resembled those of lombricine, i.e., guanidinesulfate (Guanidine), 2 guanidinoethanol sulfate (2-GEt), 4-guanidino-1-butanol sulfate (4-GBt), guanidinoethylphosphate (GEP), opheline (OPHE) and the L-form of lombricine (L Lom), were investigated in relation to their structures. In Experiment I, each guanidino compound was suspended in olive oil at the concentration of 0.3 mg/0.05 ml, and was injected subcutaneously for 10 days to SHN mice bearing spontaneous mammary tumours. 2-GEt, GEP, OPHE and L-Lom markedly inhibited mammary tumour growth, but Guanidine and 4-GBt did not. In Experiment II, the the compounds examined, 2-GEt and GEP, also significantly inhibited mammary tumour growth when given in drinking water (0.003%) for 10 days, while the effects were much weaker than those of the injected compounds. These results indicate that the existence and the length of the ethylene group are important to antitumour activity and that the site of the antitumour activity in guanidino compounds is the guanidinoethyl group. These compounds are also suggested to be more effective when administered subcutaneously rather than intragastrically. PMID- 9413172 TI - Effect of metals and their antagonists on the radical intensity and cytotoxicity of ascorbates. AB - Five heavy metal antagonists were compared for their specificity of chelating action against copper (CuCl, CuCl2) and iron (FeCl2, FeCl3). ESR spectroscopy showed that both copper and iron significantly enhanced the radical intensity of ascorbate and sodium 5,6-benzylidene-L-ascorbate (SBA). Equimolar concentrations of dimercaprol efficiently chelated all these metals, thus significantly reducing their stimulation effects. On the other hand, the chelating action of penicillamine, ethylenediaminetetraacetic acid and diethylenetriaminepentaacetic acid was limited to CuCl and CuCl2 whereas deferoxamine mesylate (DFO) was a specific iron chelator. The cytotoxic activity of sodium ascorbate was augmented by DFO, but diminished by FeCl3. The simultaneous addition of DFO and FeCl3 counteracted each other, thus neutralizing their individual effects. The cytotoxic activity of both sodium ascorbate and SBA was significantly enhanced by CuCl2 and this stimulation effect of CuCl2 was effectively chelated by DTPA. The present study demonstrates the specificity of the chelating action of these five antagonists, suggesting the possible application of these different types of antagonists for the prevention of the pathogenic diseases catalyzed by the corresponding metals. PMID- 9413173 TI - Camptothecin delivery systems: the antitumor activity of a camptothecin-20-0 polyethylene glycol ester transport form. AB - BACKGROUND: This study was designed to assess the efficacy of polyethylene glycol(PEG) conjugated camptothecin, PEG-alpha-camptothecin, a novel water soluble transport form (prodrug) of the naturally derived antitumor drug, 20-(S) camptothecin. MATERIAL AND METHODS: Circulatory retention studies were performed in non-tumor bearing mice injected intravenously with 300 mg/kg of PEG-alpha camptothecin. Therapeutic efficacy was evaluated in both a murine P388/0 leukemia and a colorectal HT-29 carcinoma xenograft model. RESULTS: PEG-alpha-camptothecin had a blood t1/2 alpha of less than 5 minutes and a t1/2 beta of 3.5 hours. Five intraperitoneal injections of 3.2 mg/kg/day 20-(S)-camptothecin equivalents of PEG-alpha-camptothecin in our leukemia model resulted in significant survival over untreated controls (P < 0.001), with a mean time to death of treated versus control (T/C ratio) of 2.94 and a cure rate of 80% (n = 20). The colorectal carcinoma xenograft model demonstrated that 2-3 mg/kg/day 20-(S)-camptothecin equivalents of PEG-alpha-camptothecin given 5 days a week for 5 weeks could reduce an initial tumor burden of 300 mm3 by more than 90% without any signs of overt toxicity. CONCLUSION: This water soluble transport form of 20-(S) camptothecin and its underlying technology may have clinical application. PMID- 9413174 TI - Bcl-2 and mdr-1 gene expression during doxorubicin-induced apoptosis in murine leukemic P388 and P388/R84 cells. AB - The induction of apoptosis by doxorubicin (DOX) alone or in the presence of efflux blockers, verapamil (VPL) or trifluoperazine (TFP), and of bcl-2 and mdr-1 gene expression was analyzed in murine leukemic P388 and doxorubicin resistant P388/R84 cells. Incubation with DOX (0.1-1 microM) for 24 hours induced apoptosis in sensitive cells but not in the resistant P388/R84 cells. Flow cytometric analysis of apoptosis by terminal dideoxynucleotidyl (TdT) assay showed that 1 microM DOX induced apoptosis in 70% of P388 cells and in less than 5% of P388/R84 cells. When P388/R84 resistant cells were co-incubated with DOX and efflux blockers (10 microM VPL or 15 microM TFP), enhanced cellular DOX accumulation was accompanied by apoptosis. Quantitative analysis of DNA fragmentation by 14C thymidine incorporation and gel electrophoresis of fragmented DNA confirmed the results from TdT assay and showed enhancement of DOX-induced DNA fragmentation in the presence of efflux blockers (VPL or TFP). DOX + VPL and DOX + TFP combination treatments induced apoptosis in upto 55% and 83% of P388/R84 cells, respectively. mdr-1 mRNA and P-gp expression were not altered during DOX-induced apoptosis. The results suggest that in murine leukemic cells, down-regulation of bcl-2 mRNA expression occurs during DOX-induced apoptosis and it depends on the cellular drug retention determined by mdr-1/P-gp drug efflux. PMID- 9413175 TI - Cellular thioredoxin reductase activity is regulated by selenium. AB - Selenium (Se) is an essential trace element and has been reported to decrease the incidence of some human cancers. We have investigated the effects of Se on thioredoxin reductase, a selenocysteine containing flavoenzyme, in HT-29 human colon cancer cells grown in serum-free medium. Sodium selenite and other Se containing compounds produced a time and concentration dependent increase in intracellular thioredoxin reductase activity and protein levels. Selenite was the most active of the Se compounds examined: 1 microM selenite produced a 28-fold increase in thioredoxin reductase activity by 1 day and 10 microM selenite over a 60-fold increase by 5 days. The activity of a related non-selenocysteine containing flavoenzyme glutathione reductase was not increased by selenite. Selenite, but not the other Se containing compounds inhibited cell growth at concentrations above 2 microM. The results show that Se can produce large increases in cell thioredoxin reductase activity. PMID- 9413176 TI - Characterisation of the G1/S cell cycle checkpoint defect in lung carcinoma cells with different intrinsic radiosensitivities. AB - Cell cycle perturbations in three lung carcinoma cell lines (U-1285,U1906 and U 1810) with different intrinsic radiosensitivities (SF2 U-1285 = 0.25, SF2 U-1906 = 0.45, SF2 U-1810 = 0.88) were investigated following x-irradiation. Cell cycle flow calculations showed that the G1-->S-phase transit was accelerated in irradiated compared with untreated U-1285 cells, up to 24 hours postirradiation. In U-1810 cells and U-1906 cells the postirradiation G1-->S transit decreased compared with controls. All three cell lines showed no postirradiation induction of p53 and p21CIP1 proteins. Cyclin E was overexpressed and cyclin E-dependent kinase activity was substantially induced by irradiation in U-1285 cells compared with U-1906 and U1810 cells while p27KIP1 was detected at the highest intensity in U-1810 cells and lowest in U-1285 cells. We hypothesise that the accelerated postirradiation G1-->S transit in U-1285 cells is associated with induction of cyclin E-dependent kinase activity and may account for increased radiosensitivity in these cells. PMID- 9413177 TI - Modulation by interferon-gamma of zinc-alpha 2-glycoprotein gene expression in human epithelial cell lines. AB - Zinc-alpha 2-glycoprotein has been detected in most body fluids, and its antibody labels the corresponding glandular epithelia. We have also detected it in human stratified epithelia (epidermis and buccal mucosa). In this study, the mRNA levels of zinc-alpha 2-glycoprotein were found to be about twice as high in epithelial cells of mucosal origin (whether normal primaries or neoplastic cell lines) as in epidermoid cells (normal epidermal primary cultures, an immortalized but non-tumorigenic epidermal cell line, and neoplastic vulvar and cervical cell lines). Interferon-gamma strongly upregulated gene expression, but substantially less in mucosal than epidermoid cells. To compare responses as a clue to the function of zinc-alpha 2-glycoprotein, we ran parallel experiments with three markers of distinct properties, all known to be induced by interferon-gamma. There was the least resemblance for involucrin, a qualitative similarity for HLA DR, and a rather better match for 2'-5' oligoadenylate synthetase. PMID- 9413178 TI - Cytoskeleton alteration in MCF7R cells, a multidrug resistant human breast cancer cell line. AB - Various cytoskeleton modifications are associated with malignant cell transformation and have been used as prognostic factors. A human breast cancer cell line (MCF7S) and its multidrug resistant (MDR) subline (MCF7R) were characterized here for their intermediate filaments (IFs) expression (cytokeratin 8, 18, 19 and vimentin) as a function of their resistance phenotype. Modifications of these cytoskeleton molecules were analyzed by flow cytometry, immunofluorescence, electrophoresis and immunoblotting techniques. Cytokeratins 8 and 18 were similarly expressed in the cell lines. Cytokeratin 19 was expressed in the MCF7S cell line and not in the MCF7R variant, while vimentin was highly expressed in MCF7R and slightly in MCF7S. Analysis of IFs after the addition of doxorubicin (Dox) in the culture medium of MCF7S, showed an increase in cytokeratin 8 filaments. Vimentin expression in MCF7R was not modified in the presence of these different MDR modulators. Acquisition of MDR was associated with an increase and a redistribution of vimentin filaments characterized by a perinuclear polarization. These drug resistance associated changes might derive from different biological processes triggered by chemotherapy. In conclusion, this suggests that this intermediate filament could be a marker associated with chemoresistance or a marker of malignancy in certain epithelial cancers. PMID- 9413179 TI - Tumor suppressive action of indomethacin is NK-cell-independent. AB - This study was undertaken to determine whether NK-cells constitute a necessary mediator for the suppression of tumor growth by indomethacin. C57Bl mice with a methylcholantrene (MCG 101) tumor were studied. Indomethacin treatment was provided by daily subcutaneous injections (1 microgram/g body weight). NK-cells were depleted by treatment with a monoclonal antibody to NK1.1. Consecutive indomethacin injections prolonged survival in tumor bearing animals. Indomethacin was equally effective in animals with intact NK-cells as in NK-cell-depleted animals. Further, the MCG cells were apparently insensitive to the lytic activity of NK-cells in vivo. Thus, the clearance of intravenously injected MCG cells from lungs was not affected by depletion of NK-cells in vivo; in contrast, the corresponding clearance of NK-cell-sensitive YAC-1 lymphoma cells was strikingly reduced by the depletion of NK-cells. Our data suggest that NK cells are not a necessary mediator for the suppression of tumor growth by indomethacin. PMID- 9413180 TI - The primary in vitro antitumor screening of "half-mustard type" phenothiazines. AB - The antitumor effects of "half-mustard type" phenothiazines were studied on 57 different tumor cell lines, including leukemias, non-small lung cancer, colon, central nervous system, ovarian, renal, breast, and prostate cancer, as well as melanoma cell cultures. Alkyl-urea derivatives of phenothiazines displayed in vitro antitumor activity. The phenothiazine phthalimido derivatives (1-6) were not active on the majority of cancer cell cultures. In contrast, propylureas (9, 11) were active against some leukemia cell types. Only two compounds with the butylene [(CH2)4] linker (10, 12) were active against non-small lung cancer cells. Compounds containing the propylene linker were less effective. On colon cancer lines, tumor cells from the central nervous system and on melanoma cells the same compounds were effective, however, having substituents at the 2-position of phenothiazine seems to be important. Surprisingly, the majority of ovarian cancer cell lines (except one type, IGROVI) and five of eight renal cancer lines were not sensitive to these phenothiazine derivatives. The two butylene linked phenothiazine ureas (10, 12) had moderate antiproliferative action on two renal cancer cell lines. The prostate cancer and some breast cancer cell lines were not sensitive. Nevertheless some breast cancer cell lines were apparently sensitive to CF3-substituted phenothiazine alkylureas. On the basis of these experiments one may postulate that in the case of insensitive cells an mdr-gene encoded multidrug resistance efflux pump is responsible for the resistance. The selectivity or organ cell specificity of the effective phenothiazines will be targeted for improvement in further studies, in order to avoid the general cytotoxic effects of "half mustard type" phenothiazines. PMID- 9413181 TI - The in vitro antitumor assay of "half-mustard type" phenothiazines in screens of AIDS-related leukemia and lymphomas. AB - Twelve different "half-mustard type" phenothiazines were newly synthesized and tested on seven AIDS-related lymphoma (ARL) tumor cell lines, one leukemia CCRF CEM cell culture and five different lymphoma lines; RL, KD-488, AS283, PA682 and SU-DHL-7 cell lines. The alkylene-urea substituted phenothiazines affected the growth and inhibited the growth rate of AIDS-related lymphoma cells. The Cl substituent at the 2-position was more effective than the CF3 substitution. In AIDS-related leukemia also the compounds with Cl at the 2-position with propylene or butylene linkers, -(CH2)3- and -(CH2)4-, respectively, were more effective than the CF3 substituted compounds. Two of the six phenothiazine-substituted alkyl-urea derivatives, i.e., 1-(2-chloroethyl)-3-(2-chloro-10H-phenothiazin-10 yl)propyl-l-urea (9, GI50 = -5.66, TGI = -5.04) and 1-(2-chloroethyl)- 3-(2 chloro-10H-phenothiazin-10-yl)butyl-1-urea (10, GI50 = -5.61, TGI = -5.12) exhibited antitumor activity for AIDS-related leukemia and five AIDS-related lymphomas. The trifluoromethyl-substituted derivatives were not as effective on AIDS-related tumor cell lines. Apparently, the substituent at the 2-position on the phenothiazine and the alkylene number of the linker attached to the nitrogen of the phenothiazine ring have an important role in the compound's antitumor effects on AIDS-related leukemia and lymphomas. PMID- 9413182 TI - Relationship between biological activity and dipole moment in benzo[a]phenothiazines. AB - Relationship between the biological activity and two different dipole moments were investigated in 9 benzo[a]phenothiazines [1-8] by calculating the dipole moment with the PM3 method. 12H-Benzo[a]phenothiazine [1], 9-methyl-12H benzo[a]phenothiazine [2], 10-methyl-12H-benzo[a]phenothiazine[3] and 11-methyl 12H-benzo[a]phenothiazine [4] induced monocytic differentiation of human myelogenous leukemic cell lines and displayed antitumor activity. These active compounds showed a significantly smaller value of calculated ground-state dipole moment (mu g) and larger value of first excited-state dipole moment (mu e). The mu e/mu g ratio of four active compounds [1-4] ranged from 3.25 to 4.38, whereas that of three inactive compounds [5,6,7] ranged from 1.77 to 2.72. These two different dipole moments might be useful parameters for estimation of the biological activity of benzo[a]phenothiazines [1-8]. PMID- 9413183 TI - DNA-protective activity of new ribonucleotide reductase inhibitors. AB - The DNA-protective activity of hydroxyurea (HU) and novel ribonucleotide reductase (RR) inhibitors amidox (AX), didox (DX) and trimidox (TX) was examined using hydrogen peroxide as the DNA-damaging agent. The exposure of superspiralized plasmid DNA molecules (pBR 322) to H2O2 under precisely defined in vitro conditions initiates a change in DNA topology (DNA from I relaxes to DNA form II). This electrophoretically monitored change in the plasmid DNA topology is related to the induction of ss-DNA breaks and corresponds with DNA exposition to free radicals. The inhibition of DNA relaxation (the prevention of DNA damage induced by hydrogen peroxide) depended on the free radical scavenging capacity of the drugs investigated. HU exerted DNA protective activity at a concentration of 4 mM, AX at concentration of 1 microM, TX at a concentration of 5 microM and DX at a concentration of 25 microM (the free radical scavenging activity increases from HU to AX in following manner: HU << DX < TX < AX). It can be concluded that the new synthetic RR-inhibitor AX which is being investigated at the preclinical level as a potential anti-cancer drug possess the highest capacity for scavenging of free radicals. PMID- 9413184 TI - Isolation of a cDNA clone from colon carcinoma. AB - cDNA clones of differentially expressed mRNAs in a colon carcinoma have been isolated by subtractive cDNA cloning. The substracted material was at least 90x enriched for differentially expressed sequences and can be used for the construction of substractive cDNA libraries and polymerase chain reaction (PCR) amplification to generate differential probes. In this way rare mRNA (less than 0.1% abundance) which are differentially expressed, can be isolated utilising this procedure. A cDNA clone which we call IH12 has been isolated. Its mRNA is expressed exclusively in actively proliferating cells. PMID- 9413186 TI - Expression of the transmembrane glycoprotein CD44 and metastasis associated 18A2/MTS1 gene in B16 murine melanoma cells. AB - CD44 is a transmembrane cell adhesion-mediating protein which occurs as alternatively spliced isoforms in a variety of cell types. CD44 isoforms have been reported to be differentially expressed in cancers and the isoform CD44v6 has often been associated with metastatic potential. The 18A2/mts1 gene, which codes for a Ca(2+)-binding protein of the S-100 family, is a metastasis associated gene and its expression has been shown to be related to cell proliferation, cancer metastasis and invasion. The association of 18A2/mts1 expression with invasion has been attributed to its ability to promote depolymerisation of cytoskeletal elements and it appears to also participate in the remodelling of the extracellular matrix. Here we have examined the expression of CD44v6 in metastatic variants of the B16 melanoma with different levels of 18A2/mts1 expression. We found that metastatic potential was not related to the overall CD44 expression as detected by immunostaining; the high metastasis variant ML8 showed reduced CD44v6 positivity as compared with the low metastasis variant F1. Up-regulation of 18A2/mts1 expression in F1 cells by a-melanocyte stimulating hormone (MSH) and its down-regulation in ML8 cells by RA reduced the numbers of CD44v6 staining cells. In F1 cells the glycoprotein was found to be uniformly associated with the cell membrane. But in F1 cells treated with a-MSH where 18A2/mts1 expression was up-regulated, CD44v6 showed redistribution into a patchy focal pattern. This patchy focal distribution of CD44v6 also occurred in ML8 cells which express 18A2/mts1 at a high level. It is suggested that the patching of CD44v6 molecules is a consequence of the changes in cytoskeletal dynamics brought about by 18A2/mts1 expression, that are conducive to aggregation and patching of these transmembrane glycoproteins. It is postulated that this induction of patching could provide discrete and strong adhesive foci promoting cell adhesion and invasive behaviour. PMID- 9413185 TI - Mechanism of apoptosis induced by cisplatin and VP-16 in PANC-1 cells. AB - Cisplatin and VP-16 were used to study the induction of apoptosis in Panc-1 cells. Cisplatin and VP-16 inhibited the growth of Panc-1 cells. After 2 hours exposure to cisplatin or VP-16, attached and detached cells were subjected to TUNEL staining to calculate the ratio of apoptosis. In detached cells TUNEL positive ratios increased in a time- and dose-dependent manner. In Western blotting, Bax expression was obviously up-regulated, but Bcl-2 remained almost constant. The results suggested that in Panc-1 cells cisplatin and VP-16 induced apoptotic cell death which was mediated through the interaction of Bax expression in the presence of mutated p53. PMID- 9413187 TI - Modifying effect of organocobalt complexes on the tumour response to anticancer treatments. AB - A new type of agents are proposed for combined cancer therapy. They are organocobalt (III) chelates containing a sigma-bounded organyl group and a mixed tridentate ligand derived from a Schiff base. These complexes generate free radicals due to the action of protons in physiological ranges of pH and temperature, and hence are conceivably capable of selectively attacking a malignant neoplasm that is slightly acidic and can be made even more so by introducing some means intensifying glycolysis. An in vivo examination was performed using transplanted rat tumours (Guerin and Walker 256 carcinomas, Sarcoma 45). The modifying effect of one of these complexes on the tumour response to cis-DDP, radiation and/or local hyperthermia was tested by means of tumour growth delay assay and local tumour control. The potentiating effect of the complex was maximal when it was administered 60-90 minutes prior to other agents (cisDDP, X-irradiation heat). The enhancement ratio was found to be ca. 2.0-4.0 for cisDDP and 2.0 for radiation. In conclusion, in our tumour models, an increase of the antitumour effect was obtained for conventional antitumour agents when they were supplemented with organocobalt complex. It can be hypothesised that DNA in tumour cells may be considered to be the main target for organocobalt complexes. PMID- 9413188 TI - In vivo overexpression of c-erbB-2 oncoprotein in xenografts of mice implanted with human colon cancer lines. AB - We have previously shown that c-erbB-2 oncoprotein encoded by the erbB-2 gene is overexpressed in human colorectal cancers that metastasis compared to those that are cured by surgery. To determine whether c-erbB-2 is also differentially expressed in vivo in metastasising and non-metastasising tumours, we developed models of colorectal cancer growth in nude mice. Human colon cancer cell lines, HCT116, KM12SM, LIM1215 and SW480, were injected into the caecum after characterising their morphology, doubling time, DNA flow-cytometry and expression of c-erbB-2. Six weeks later, xenografted tissues were fixed for histological analysis and detection of c-erbB-2 by immunohistochemistry, 78% (21/27) of mice developed caecal cancers. The caecal tumours derived from HCT116, KM12SM or LIM1215 were highly metastatic; 67 to 100% of them had liver metastases and lymph node involvement and 33 to 75% had lung tumours. Most of the tumours were c-erbB 2-positive. In contrast SW480 caecal tumours had only 33% lymph node involvement, but not liver or lung metastases. Only one SW480 caecal tumour and one lymph node metastasis expressed c-erbB-2. C-erB-2 was more frequently expressed in xenografted tissues in colon cancer primaries and secondaries of the highly metastatic cells (HCT116, KM12SM and LIM1215) compared to the cells (SW480) giving predominantly local growth. Our results suggest that c-erbB-2 gene may play an important role in the development of metastasis from colorectal cancer. PMID- 9413189 TI - Cisplatin and mild hyperthermia in radiosensitization to low dose rate irradiation in human ovarian carcinoma cells. AB - Two human ovarian carcinoma cell lines, one parental and the other a cisplatin resistant derivative cell line, were evaluated for the efficacy of combined treatments of mild hyperthermia, cisplatin and low dose rate irradiation (LDRI). The data showed that cisplatin (1 or 3 micrograms/ml for 1 hour) combined with mild hyperthermia (40 degrees C for 1 hour) was effective for radiosensitization to LDRI in the parental cell line whether given before or after irradiation. In the cisplatin resistant variant, however, these treatments had little to no effect on LDRI response. Thus these data show that these treatments could be clinically effective if there is no cisplatin resistance. Our previous study showed that concomitant treatments could be effective in both parental and cisplatin resistant cell line (1). PMID- 9413190 TI - Conformational changes in gastric carcinoma cell membrane protein correlated to cell viability after treatment with adamantyl maleimide. AB - BACKGROUND: In order to investigate the mechanism of the effects of the anticancer drug adamantyl maleimide (AMI) on the in vitro growth of a human gastric carcinoma cell line (SC-M1), the membrane protein conformation on the cell surface after treatment with AMI was studied. MATERIALS AND METHODS: The interaction of AMI with SC-M1 cell line was studied by reflectance Fourier transform infrared (FT-IR) microspectroscopy. The cell viability was also determined by the trypan blue exclusion method. RESULTS: FT-IR microscopic study provided evidence that the conformational conversion of the membrane proteins of the SC-M1 cell line from alpha-helix to beta-sheet was found evident, but was dependent on the AMI concentrations used and the time of treatment. AMI not only interacted with the membrane protein on the cell surface, but also entered into cells to interact with the nucleic acids. CONCLUSIONS: The relationship between membrane protein conformational changes and the cell viability of SC-M1 cell line was evident. The AMI-induced apoptosis in SC-M1 cell line was also supposed. PMID- 9413191 TI - Rapid determination of ascorbate by ESR spectroscopy. AB - Optimal conditions for the determination of ascorbate, based on the detection of its radical by ESR spectroscopy, were established. The optimal pH for the measurement of the ascorbate radical was 10, whereas that for the gallate and caffeate radical was much higher. This was due to the liability of the ascorbate radical at higher pH. When the radical intensity of ascorbate vs. concentration was plotted double logarithmically, a linear correlation was established between these two parameters within the ascorbate concentration of 0.03-10 mM. ESR analysis of the homogenates under alkaline condition demonstrated that ascorbate is distributed in various mouse organs and that the ascorbate radical is unstable at neutral pH in the brain and liver, suggesting the presence of metabolizing substances. The present study suggests the applicability of the present technique for the study of ascorbate metabolism and identification of physiological factors which regulate the radical intensity of ascorbate. PMID- 9413192 TI - Comutagenesis-I: the in vitro metabolism of 2-amino-3-methylpyridine. AB - The metabolism of the comutagen 2-amino-3-methylpyridine has been studied in vitro using rat and rabbit hepatic preparations. 2-Amino-3-methylpyridine-N oxide, 2-amino-3-hydroxymethylpyridine and 2-amino-5-hydroxy-3-methylpyridine were formed by both rat and rabbit hepatic preparations. No evidence was obtained for the formation of the corresponding 2-hydroxylamine, 2-nitroso, 2-nitro-3 methyl-pyridine or their condensation products i.e. azo, azoxy or hydrazo. The results are discussed in relation to the possible mechanism of action of the substrate. PMID- 9413193 TI - Multidrug resistance-associated protein gene expression and drug sensitivity in human lung cancer cells. AB - Multidrug resistance-associated protein (MRP) mRNA expression and drug sensitivity in lung cancer cells were examined, and the effects of verapamil, a modulating agent for MRP, on drug sensitivity were also tested. Nine cell lines expressed various levels of MRP gene expression but not the MDR1 gene. The levels were higher in non-small cell carcinoma cells (NSCLC) than in small cell carcinoma cells (SCLC). Clear correlations between the MRP gene level and the sensitivity to etoposide (VP-16) and doxorubicin (Dox) were observed except for one cell line which highly expressed DNA topoisomerase II. Positive correlations between the MRP gene levels in three cell lines and the modulation effects of verapamil in VP-16, Dox, and vincristine were observed. The present results indicate that MRP probably confers intrinsic multidrug resistance in NSCLC rather than in SCLC. PMID- 9413194 TI - Comparison of the effects of epidermal growth factor and fetal calf serum in human endometrial carcinoma RL95-2 cells. AB - This study was designed to evaluate if c-myc expression could influence RL95-2 cell proliferation after EGF or fetal calf serum (FCS) stimulation. Upon FCS addition, c-myc mRNAs slightly increased in the first 30 minutes, peaked at 75 minutes (2.2 fold induction) and returned to the control value within 24 hours. This slight and transient FCS effect on c-myc expression contrasted with the marked and long-lasting EGF effect (17 fold induction up to 48 hours). The increase of cell number by FCS was partially but significantly reduced in the presence of c-myc antisense oligodeoxynucleotides (ODNs). The EGF inhibitory effect on cell growth was not reversed by c-myc antisense ODNs whatever the backbone may be (phosphorothioate or phosphodiester). It appears that EGF and FCS have differential effects on c-myc and RL95-2 cell proliferation and that c-myc is necessary but insufficient for proliferation. PMID- 9413195 TI - The association of sialyl Lewis(a) antigen with the metastatic potential of human colon cancer cells. AB - BACKGROUND: The vascular endothelium plays a critical role in cancer metastasis by directing circulating cancer cells into extravascular tissues and by expressing cell adhesion molecules. The authors investigated the interaction between a cell line derived from human colon cancer and cultured murine endothelial cells, in vitro, and in vivo. MATERIALS AND METHODS: Variant cell lines with high (WiDr-HA) or low (WiDr-LA) levels of cell surface sialyl Lewis(a) antigen (s-Le(a)) were isolated from the heterogeneous WiDr cells (WiDr-P) in order to characterize the biological behavior of colon cancer cells having elevated cell surface contents of s-Le(a). RESULTS: A correlation was found to exist between the degree of s-Le(a) expression, and the attachment of cancer cells to activated human umbilical vein endothelial cells, or to F-2 cells isolated from murine vascular endothelial cells. The adhesion of WiDr-P and WiDr HA to F-2 cells was inhibited by the addition of anti-s-Le(a) antibody (Ab). WiDr P and WiDr-HA both demonstrated the capacity to metastasize to the liver, by the inoculation of cancer cells to the spleen, while WiDr-LA did not do so. The number of metastatic nodules of WiDr-HA was significantly higher than that of WiDr-P. Treatment with anti-s-Le(a) Ab inhibited the metastasis of WiDr-P and WiDr-HA to the liver, but not with anti-s-Le(x) Ab. CONCLUSIONS: These findings suggest that s-Le(a) plays an important role in cell adhesion molecules, in the metastasis of colon cancer. PMID- 9413196 TI - Modulating factors of radical intensity and cytotoxic activity of ascorbate (review). AB - Ascorbate acts both as an antioxidant and as an oxidant, depending upon the environment in which the molecule is present. We have reported that millimolar concentrations of ascorbate induced apoptotic cell death, characterized by cell shrinkage, nuclear fragmentation and internucleosomal DNA cleavage, in human myelogenous leukemic cell lines. Ascorbate derivatives, which can induce the apoptosis, produced the radical(s), elevated the oxidation potential and stimulated the methionine oxidation in the culture medium, whereas inactive derivatives did not. This suggests that the ascorbate induce the apoptosis by its prooxidant action. The effects of various factors, such as temperature, pH, metal, metal antagonist, redox agent, serum protein, polyphenol and (natural, chemically modified) polysaccharide on the radical intensity and apoptosis inducing activity of ascorbate are reviewed. Gallate and benzo[a]phenothiazine derivatives, which can induce apoptosis or monocytic differentiation in human myelogenous leukemic cell lines, also produced radicals. These data suggest the significant role of radicals in the initiation of diverse biological activities. PMID- 9413197 TI - Targeted vinblastine chemotherapy with two preparations of lipiodol contrast medium. AB - BACKGROUND: Long-term survival and cure can not be achieved in patients with unresectable metastatic liver cancer. In this study, to improve the antitumor activity of oily anticancer agents, which enable targeted cancer chemotherapy, various concentrations of vinblastine dissolved in Lipiodol ultrafluid (vinblastine/Lipiodol U) or Lipiodol F, which has three times the viscosity of Lipiodol ultrafluid, were used. MATERIALS AND METHODS: Rabbits bearing VX2 tumors measuring 1 to 2 cm in diameter in the liver received arterial injections of 0.2 ml of these drugs, and antitumor activity were examined. RESULTS: Complete necrosis of the tumor was observed in three of nine rabbits received vinblastine/Lipiodol U at 2.5 mg/ml. Complete necrosis of the tumor was observed in all eleven rabbits which received vinblastine/Lipiodol U at 5 mg/ml. At concentrations of 7.5 and 10 mg/ml, the necrotic area spread from the tumor to the liver parenchyma surrounding the tumor. The antitumor activity of vinblastine dissolved in Lipiodol F was increased compared with that of vinblastine dissolved in Lipiodol ultrafluid. CONCLUSIONS: The antitumor activity of a kind of the oily anticancer agents, vinblastine/Lipiodol increased with increasing amounts of vinblastine dissolved in Lipiodol ultrafluid and with the increasing viscosity of the carrier, Lipiodol. PMID- 9413198 TI - Characteristics of human gastric carcinoma cell lines with induced multidrug resistance. AB - In contrast to intrinsic drug resistance, induced multidrug resistance in gastric cancer cells has not been well studied. Therefore, two doxorubicin-resistant cell lines, (SNU-1DOX, SNU-16DOX), were derived in vitro from gastric carcinoma cell lines (SNU-1, SNU-16) respectively, and their characteristics were investigated. These resistances were not associated with overexpression of mdrl, multidrug resistance associated protein 1 (MRP1), pi or liver class of glutathione S transferase (GST pi, GSTL), heat shock protein 70 (HSP70), p53 or transglutaminase C (TGC). Levels of p21WAF1 RNA and topoisomerase II protein were decreased in the SNU-16DOX, but not in SNU-1DOX. However, the subsequent enzyme activity of topoisomerase II in SNU-16DOX was not decreased, but rather increased in SNU-16DOX. Furthermore, both resistant cell lines showed lower uptake and higher efflux of doxorubicin and induced cross-resistance to etoposide and vincristine in addition to doxorubicin, indicating a multi-drug resistance phenotype. In summary, we report two gastric carcinoma cell lines exhibiting induced multidrug resistance phenotype and suggest that mdrl, MRP1, GST, TGC, HSP70 and p53 do not play important roles in induced drug resistance in these cell lines. The role of changes in topoisomerase II activity and/or protein is still inconclusive, and p21WAF1 is associated with induced multidrug resistance in the SNU-16DOX gastric carcinoma cell line. PMID- 9413199 TI - Drug resistance reversal, anti-mutagenicity and antiretroviral effect of phthalimido- and chloroethyl-phenothiazines. AB - The effect of substituted phenothiazines was studied in three different systems; bacteria and cancer cells and reverse transcriptase enzyme of Moloney leukemia virus. F'lac and hemolysin plasmids were eliminated by some substituted phenothiazines from E. coli at a very low frequency. The same phenothiazine derivatives also were synergistic with tetracycline in bacteria and shown antimutagenic effect in Ames test. No mutagenic effects were observed in TA 98 strain of Salmonella typhimunium. Chloroethyl-substituted phenothiazines showed antimutagenicity equivalent to the parent compounds; however, phthalimido substituted phenothiazines had higher antimutagenicity of 50%. P-glycoprotein responsible for multidrug resistance was also inhibited in tumor cells. The accumulation of the fluorescent rhodamine 123 in the phenothiazine treated multi drug resistant tumor cells was measured by flow cytometry. Some of the substituted phenothiazines were effective P-glycoprotein blockers, while some compounds had moderate activity, but others were without effect as compared to 5 microM verapamil. On the basis of computer analysis there are some correlations between the biological activities and the dipole moments, and entropy of the studied molecules. Our results suggest that the inhibition of Hly+ plasmid replication and P-glycoprotein function may depend partly on similar electronic properties of the studied phenothiazine derivatives. The activity of Moloney leukemia virus reverse transcriptase was inhibited by the substituted phenothiazines, however, no basic differences were found in the activities of phthalimido- and chloroethyl substituted phenothiazines. PMID- 9413200 TI - p53, c-erbB-2 and p21ras expression in tumor effusion cells of patients with histopathologically different ovarian neoplasms. AB - The overexpression of p53, c-erbB-2 and p21ras gene products was evaluated immunohistochemically in ovarian carcinomas, borderline, and benign neoplasms. All studies were performed on cytospin preparations of cyst and/or ascitic fluid cells and mutual relations between oncoproteins were analysed. p53 and c-erbB-2 immunostaining was observed in 50% and 48.5% of ovarian carcinomas and in 30% and 35% of ovarian borderline tumors respectively, however in the last group the intensity and percentage of stained cells were considerably lower. In ovarian benign neoplasms there was no evidence of p53 and/or c-erbB-2 expression. The trend for serous carcinoma to have a higher p53 and c-erbB-2 expression than endometrioid and mucinous carcinomas was observed. p53 and c-erbB-2 oncoproteins were detected more frequently in the III/IV than in the I/II stages of the disease. The expression of p21ras was detected in 91% of malignant, 65% of borderline and 50% of benign neoplasms. p21ras reactivity was independent of the histopathological structure of ovarian carcinomas and it was comparable in I/II and III/IV FIGO stages. Our results indicate that p21ras overexpression appears to be an early genetic alteration in ovarian tumorigenesis, followed by the appearance of p53 and c-erbB-2 oncoproteins. It is likely that enhanced p53 and c erbB-2 expression may cooperate with ras gene activation to produce a particularly aggressive phenotype. Our study supports the concept that development of ovarian carcinoma is the end result of a complex multistep process involving the complementary action of different cancer causing genes. PMID- 9413201 TI - Radical intensity and carcinogenic activity of benz[c]acridines. AB - Among 13 benz[c]acridines, six 7-methyl-substituted compounds (7 methylbenz[c]acridine, 7,9-dimethylbenz[c]acridine, 7,10-dimethylbenz[c]acridine, 7,11-dimethylbenz[c]acridine, 7,9,10-trimethylbenz[c]acridine, 7,9,11 trimethylbenz[c]acridine) were carcinogenic, while the other seven compounds (benz[c] acridine, 8-methylbenz[c]acridine, 9-methylbenz[c]acridine, 10 methylbenz[c]acridine, 11-methylbenz[c]acridine, 5,7-dimethylbenz[c]acridine, cis 5,6-dihydroxy-5,6-dihydrobenz[c]acridine) were inactive. Using both McLachlan Huckel molecular orbital (McLachlan-HMO) and HMO methods, all the carcinogenic compounds were shown to have the elevated pi-spin density at 12th nitrogen atom of their molecules, as compared with noncarcinogenic compounds. Electron spin resonance (ESR) spectroscopy, however, revealed that both carcinogenic and noncarcinogenic compounds produced no detectable amounts of radical. This is in contrast to ascorbates, gallates and benzo[a]phenothiazines, which induced apoptosis by radical mediated mechanism(s). Amino acid analysis demonstrated that methionine oxidation is not involved in the induction of carcinogenic activity by benz[c]acridines. PMID- 9413202 TI - Expression of the MAGE gene family in human gastric carcinoma. AB - MAGE genes code tumor antigens that are recognized by cytolytic T lymphocytes and have been shown to be expressed in various malignant tumors. However, there is still little information on the expression of the MAGE gene family except for reports of MAGE-1 and -3. In this study, we therefore investigated the expression of MAGE-4, -6, -8, -9, -10, -11 and -12, as well as MAGE-1, -2 and -3 in both cell lines and surgical samples of gastric carcinoma, using reverse transcriptionPCR. Of the investigated 11 cell lines, MAGE-4, -6, -8, -9, -10, -11 and -12 were detected in 8 (73%), 6 (55%), 2 (18%), 59 (44%), 6 (55%), 4 (36%), and 7 (64%), respectively. No expression of these genes was seen in any of the 54 samples of normal gastric tissue. In contrast, the tumor tissue samples were found to express MAGE-4, -6, -8, -9, -10, -11, and -12 in 18 (33%), 13 (24%), 6 (11%), 10 (19%), 5 (9%), 13 (24%), and 10 (19%), respectively. Forty-four (82%) of 54 gastric tumors expressed at least one of these genes. No significant correlation was observed between the expression of MAGE genes and any specific clinicopathological factors. These results may hopefully prove to be useful in developing strategies for tumor-specific immunotherapy of gastric carcinoma using MAGE gene products. PMID- 9413203 TI - Preferential cytotoxicity to tumor cells of 3,5-diprenyl-4-hydroxycinnamic acid (artepillin C) isolated from propolis. AB - A tumoricidal substance was isolated from Brazilian propolis as guided by cytotoxicity assay on HuH 13 (human hepatocellular carcinoma) cell and was characterized to be 3-[4-hydroxy-3,5-bis (3-methyl-2-butenyl) phenyl]-2-propenoic acid (3,5-diprenyl-4-hydroxycinnamic acid (artepillin C)). It exhibited preferential cytotoxicity to tumor cells cultured in vitro. The cytotoxicity observed seemed to be partly attributable to apoptosis-like DNA fragmentation. The compound showed anti-tumor activity more effective than that of 5 fluorouracil to transplantable human tumor cell lines when tested on histoculture drug response assay system. PMID- 9413205 TI - Effect of hepatocyte growth factor on cell cycle and c-met expression in human gastric cancer cells. AB - Hepatocyte growth factor (HGF) was found to stimulate the growth and progression of gastric cancer cells through hepatocyte growth factor receptor (HGFR). In the present study, the effects of HGF on the expression of HGFR in relation to cell cycle progression of human gastric cancer cells were investigated by two parameter flow cytometric analysis. We found that the expression of HGFR in SC-M1 and KATO-III gastric cancer cells was cycle dependent, the level of HGFR increased from GO-G1 to S phase and the highest level of HGFR was found in G2-M phases. The level of HGFR was higher in KATO-III than SC-M1 cells. However, HGF treatment induced a dose-dependent stimulation of growth as well as down regulation of HGFR in SC-M1 cells but not in KATO-III cells. These results suggest that functional HGFR rather than overexpressed HGFR may be more important for the growth of gastric cancer cells. PMID- 9413204 TI - Expression of nm23-H1 in human meningioma cells. AB - The expression of nm23-H1 has been demonstrated to be highly correlated with the metastatic potential of various tumors. In the present investigation, meningiomas of different pathological grades were used to study on their nm23-H1 expression. Immunohistochemistry showed that nm23-H1 was expressed mainly in the cytoplasm especially in the perinuclear region in explants under short-term culture. Western-blotting demonstrated the specific expression of nm23 protein in all tumor samples. The expression was also found to be sex-dependent on tumor progression in female, but not in male patients. RT-PCR results confirmed nm23-H1 expression was higher in benign tumors than in their normal counterpart. Our observations thus suggest that nm23-H1 may play an important role in the progression of meningiomas in female patients. PMID- 9413206 TI - Electrophoretic analysis of nuclear matrix proteins in human hepatocellular carcinoma. AB - The nuclear matrix is the non-chromatin skeleton of the nucleus. This structure contributes to the shape of the nucleus and regulates various nuclear functions. In this study, nuclear matrix proteins of human normal liver, a liver cancer cell line, HepG2, and hepatocellular carcinomas (HCC) were investigated. Using high resolution two-dimensional polyacrylamide gel electrophoresis, the nuclear matrix proteins of 3 normal liver and 14 HCC were compared and contrasted. A high degree of similarity between normal liver, HepG2, and HCC nuclear matrix protein patterns was found. Two HCC specific nuclear matrix proteins were identified. Among these, one protein (HCC-1, Mr 62 kd, pI 5.3) appeared in all tumor samples and HCC-2 (Mr 33.25, pI 5.3-5.5) was present in 9/11 tumors, but absent in normal liver and HepG2. Our results indicate the presence of HCC specific nuclear matrix proteins. These matrix proteins may be used as markers for HCC. PMID- 9413207 TI - c-Ha-ras transfection and expression of MDR-related genes in MCF-10A human breast cell line. AB - BACKGROUND: We studied the relationship between ras transfection and the drug resistance phenotype in the MCF-10A cell line (a non-transformed immortalized human breast epithelial cell line). MATERIALS AND METHODS: Parental (MCF-10A) and c-Ha-ras transfected (MCF-10A H) cell lines were tested for resistance to Doxorubicin, Maphosphamide and Cisplatinum, by evaluating the inhibition of [3H] thymidine incorporation. Cells were also examined for the expression of different resistance mechanisms by immunocytochemical and RT-PCR methods. RESULTS: ras transfected cells were much more resistant to Cis-platinum than parental ones. GST-pi the GST isoenzyme frequently involved in human cancers) increased only in transfected cells. No expression of any other resistance mechanisms (MDR, MRP, LRP) was found by immunocytochemistry either in parental or in transfected cells; nevertheless, the more sensitive RT-PCR tests detected mrp products in parental and in transfected cells: gene expression levels were low and equivalent in both cell lines. CONCLUSIONS: ras transfection can induce resistance to Cis-platinum by increasing GST-pi expression. PMID- 9413208 TI - In vivo effects of cyclophosphamide on oncogene and suppressor gene expression in a "follow up" study. AB - Cyclophosphamide is a chemotherapeutical drug, with proved carcinogenic side effects. Our previous experiments showed its effect on the expression of certain oncogenes and tumor suppressor genes. In order to further explore the effects of cyclophosphamide at the gene level an in vivo mouse model was developed. We compared the effects of cyclophosphamide with that of cyclosporine, which is a non genotoxic human carcinogenic chemical. After different time periods of intraperitoneal cyclophosphamide or cyclosporine injection, RNA was isolated from whole organs of experimental animals, and expression of onco/suppressor genes was determined by slot blot hybridisation. In our model we have proved that gene expression changes caused by cyclophosphamide can be detected even six months after treatment without tumor development. Our results support the hypothesis that gene expression investigations could be useful biomarkers in monitoring the effects of environmental carcinogenic compounds. PMID- 9413209 TI - Nanoerythrosomes, a new derivative of erythrocyte ghost: III. Is phagocytosis involved in the mechanism of action? AB - We have previously developed a new drug carrier, named nanoerythrosome which is prepared by extrusion of erythrocyte ghosts to produce small vesicles having an average diameter of 100 nm. Daunorubicin (DNR) conjugated to these nanoerythrosomes has a higher antineoplastic index than the free drug. Moreover, since nanoerythrosomes are particles, phagocytosis may be involved in their mechanism of potentiation. In the present study, we have compared the mechanism of penetration between free DNR and conjugate DNR linked to nanoerythrosomes, on cells presenting high phagocytic activity, macrophages, and cells lacking phagocytic activity, the P388 D1 cell line. Our results demonstrate that: 1) The nanoerythrosome-DNR complex is rapidly adsorbed and phagocytosed by the macrophages, but not by the P388 D1 cell line. 2) On the contrary, DNR enters both phagocytic and non phagocytic cells. Furthermore, the cellular distribution of DNR is the same in both cell lines, the nucleus being the target organelle. We conclude that phagocytosis of the nanoerythrosome-DNR complex is not involved in its mechanism of action. PMID- 9413210 TI - A quantitative study of radiation-induced apoptosis in two rat yolk sac tumour cell lines with different radiosensitivities in vitro. AB - Two rat yolk sac tumour cell lines with different radiosensitivities were used to quantify the extent of apoptosis following irradiation by using a DNA fragmentation assay in vitro. Apoptosis was also confirmed by fluorescence analysis of nuclear morphological changes by using Hoechst 33258. A radiosensitive cell line, NMT-1 cells, showed morphological changes characteristic of apoptosis by fluorescence microscopic observation at 24 hours after irradiation with a single dose of 10 Gy. Development of apoptosis in NMT-1 cells was observed as a function of time within 24 hours after irradiation. There was a significant increase in the amount of apoptosis between 2 and 5 Gy only in NMT-1 cells but increasing the radiation dose from 5 Gy to 10 Gy did not result in increased apoptosis. A radioresistant NMT-1R cells, on the other hand, displayed a small apoptotic response to an irradiation dose of 10 Gy at 24 hours after irradiation. PMID- 9413211 TI - Enhancement of cytotoxic activity by synthesis of peptide multimeric forms. AB - Synthesis of four multimeric H-Lys-His-His-Arg-Lys-Lys-His-Arg-Lys-Arg-Lys-His His-Lys-Arg-Lys-oH peptides containing two, four, eight and sixteen branches was carried out by solid phase utilizing a lysine core matrix. These multimeric peptides enhanced activity by inhibiting the colony-forming ability of HeLa cells, from twenty-four to fifty-six times in comparison with the monomeric form. Unexpectedly the peptide with only two-branched sequences showed the highest inhibitory activity. PMID- 9413212 TI - Antitumor activity of DA-125, a novel anthracycline, in human gastric and pulmonary adenocarcinoma cells resistant to adriamycin and cisplatin. AB - The study was undertaken to investigate the antitumor activity of DA-125, a promising new analogue of adriamycin (ADM) containing fluorine, against human gastric and pulmonary adenocarcinoma cell lines and their sublines resistant to ADM and cisplatin (CDDP) by MTT assay. The cells used were MKN-45, human gastric adenocarcinoma, PC-14, human pulmonary adenocarcinoma, and their sublines resistant to ADM, MKN/ADM and PC/ADM, and CDDP, MKN/DDP and PC/DDP. MKN/ADM and PC/ADM were 12.6- and 13.9-fold resistant to ADM, respectively, compared to the respective parental cell lines in terms of IC50. The survival was less in cells treated with DA-125 than that treated with ADM in all lines tested. The antitumor activity of DA-125 was compared with that of ADM using IC50 values and relative resistance (RR). RR was defined as the ratio of the IC50 value of the resistant subline to that of the parent line. In all lines tested, the mean IC50 values for both DA-125 and ADM were lowest in the parent cells, followed by CDDP-resistant cells and ADM-resistant cells. The IC50 values for DA-125 were lower than those for ADM in all six lines tested, although statistical significance was observed in four lines. ADM-resistant sublines were highly resistant to DA-125 (RRs for ADM and DA-125; 12.6 and 7.6 MKN/ADM and 13.9 and 10.3 in PC/ADM, respectively). On the other hand, CDDP-resistant sublines were also resistant to ADM and DA-125 when compared with the respective parent cell lines (RRs for ADM and DA-125; 2.3 and 2.0 in MKN/DDP and 4.8 and 6.0 in PC/DDP, respectively). Although DA-125 showed cross-resistance to ADM and CDDP, we recommended DA-125 to be a good candidate for further development because DA-125 exhibited remarkable antitumor activity against the parent cell lines. PMID- 9413213 TI - Influence of cepharanthin on the intracellular accumulation of adriamycin in normal liver cells and spleen cells of mice in vitro and in vivo. AB - Cepharanthin enhances the cytotoxicity of adriamycin by inhibiting its efflux incorporated into cancer cells. Therefore, it may also enhance accumulation of adriamycin, and then enhance toxicity by adriamycin in normal cells as well as in cancer cells. In this study, we first examined influence of cepharanthin on the accumulation of adriamycin in normal liver cells or spleen cells in vitro and in vivo. Cepharanthin lowered the accumulation of adriamycin in liver cells, in both in vitro and in vivo. In spleen cells, it enhanced adriamycin accumulation slightly in vitro, but did not affect that in vivo. Further, cepharanthin lowered the amount of adriamycin in plasma. This suggests that decreased levels of adriamycin in plasma may be one of factors in the decrease of adriamycin accumulation in liver cells. On the other hand, cepharanthin enhanced the NK 2280 accumulation used for an indicator of cell membrane potential for 60-100 minutes in liver cells, but did not change that in spleen cells. These suggest that the cell membrane potential may not affect the adriamycin accumulation in both cells since adriamycin accumulation increases with increasing accumulation of NK-2280. Moreover, no change in adriamycin efflux of liver cells was demonstrated by the treatment of cepharanthin. This indicates that the decrease in adriamycin accumulation in liver cells may not be due to the inhibition of adriamycin efflux. In conclusion, cepharanthin enhances adriamycin accumulation in cancer cells, but lowers its accumulation in normal liver cells and does not affect that in spleen cells. Therefore, cepharanthin may be useful in cancer chemotherapy. PMID- 9413214 TI - Mitomycin C and cisplatin increase survival in a human pancreatic cancer metastatic model. AB - Pancreatic cancer is one of the most intractable of all human cancers. We have previously developed a patient-like model of human pancreatic cancer by surgical orthotopic implantation (SOI). After SOI of the human tumor xenograft PAN-12-JCK into the tail of the nude mouse pancreas, mitomycin C (MMC) and cisplatin (DDP) were administered intraperitoneally at a dose of 4 and 6 mg/kg, respectively, on day-7. The mice were observed for 95 days. There was a statistically significant increase in disease-free and overall survival rates in the MMC- and MMC + DDP treated groups. Local tumor growth was eliminated only in the group treated with MMC + DDP. Hepatic metastasis and peritoneal disseminations were completely inhibited by MMC but not DDP. This study demonstrates the usefulness of the SOI model of pancreatic cancer to study the differential efficacy of agents affecting primary tumor growth metastasis and survival, thus presenting an opportunity for the discovery of new agents for this highly resistant cancer. PMID- 9413215 TI - Small cell lung cancer can express CD34 antigen. AB - In humans CD34 is a valid and reliable marker for hematopoletic stem and progenitor cells. In general, solid tumors, with the exception of endothelial cancers, do not express CD34. Accordingly, immunological selection of CD34+ hematopoietic stem/progenitor cells can be used to remove CD34- malignant cells in the setting of autotransplantation. To rule out CD34 expression on tumor cells from small cell lung cancer (SCLC), eleven SCLC cell lines were analyzed by flow cytometry. Interestingly, two of these were positive for CD34 and their expression of full-length CD34 was further confirmed by reverse transcriptase and polymerase chain reaction (RT-PCR). This finding indicates that prior to subjecting SCLC patients to the above treatment modality, preparing CD34+ hematopoietic stem/progenitor cells from SCLC patients for autotransplantation, CD34 expression on their tumor cells should be screened using immunohistochemistry and/or flow cytometry. PMID- 9413216 TI - Expression of CD44 standard and isoforms in human breast cancer xenografts and shedding of soluble forms into serum of nude mice. AB - Standard CD44 (CD44s) and variant isoforms (CD44v) are expressed on different malignant cells and tissues. Their upregulation has been implicated, in the progression and metastasis of malignomas. In this work we addressed the question of whether these molecules are also expressed on xenografted human breast carcinomas and if certain expression patterns are correlated with biological parameters like tumour size, hormone receptor status, histology, growth rate, chemoresistance and microenvironment. Additionally, we were interested in the shedding of soluble CD44 (sCD44) into the blood circulation of tumour bearing nude mice. The human breast carcinomas MCF-7, MCF-7/ADR, 4296 and MDA-MB435, 4134 and 4151 were transplanted subcutaneously (sc.) or into the mammary fat pad (mfp.) of nude mice. The expression of the CD44s, -v6, and -v9 isoforms was determined at different time points on tissue samples by immunohistochemistry or RT-PCR employing human-specific antibodies or primers, respectively. The serum concentration of CD44s and -v6 was measured by human specific ELISAs. All tumours expressed CD44s. The lowest level was observed in the MCF-7 cancer. The CD44v6 and -v9 sequences and epitopes were distinctly expressed in MCF-7/ADR. MDA-MB435, 4134, 4151 and 4296, while MCF-7 lacked these isoforms. The highest serum concentration of the v6 isoform was detected in mice bearing the tumour 4296 with a high tendency for lymphogenic metastasis. The serum levels of sCD44 were in 5/6 xenografts linearly correlated with the tumour size. Interestingly, there was a remarkable difference between the two sublines MCF-7 and MCF-7/ADR: Both the tissue and serum levels of CD44 isoforms indicated that the development of multidrug resistance is accompanied by an alteration in the expression of membrane proteins discussed to be involved in metastasis. There was no relation of tissue expression with the transplantation site and the hormone receptor status of the tumour lines. CD44s and its variant isoforms are expressed in human xenotransplanted breast cancers in very different levels and patterns. The highest expression in the tumour 4296 is related to lymphogenic metastasis while the absence of isoforms in the model MCF-7 is related to non-metastatic behaviour. CD44 is shed into the serum and can be used for monitoring of tumour growth. PMID- 9413217 TI - Combination therapy of the H2712 murine mammary carcinoma with cytotoxic T lymphocytes and anti-estrogens. AB - BACKGROUND: The triphenylethylene antiestrogenic agents, tamoxifen (TX) and toremifene (TO), are currently being used for the therapy of estrogen dependent breast carcinomas and for some other estrogen receptor positive tumors. Some observations indicate that these drugs may have a beneficial effect on estrogen receptor negative tumors as well. MATERIALS AND METHODS: The H2712 mammary carcinoma of C3H/HeJ mice was studied in combination immunotherapy experiments using cytotoxic T lymphocytes (CTL) and TX or TO. The effect of TX and TO on the in vitro lysis of H2712 cells by CTL was also investigated. Finally, the H2712 cells were examined for the presence of estrogen receptors. RESULTS: Both TX and TO potentiated the lysis of H2712 cells by CTL in vitro, and exerted a growth inhibitory and therapeutic effect on H2712 mammary carcinomas in vivo. The therapeutic effect of CTL isolated from tumor bearing mice was improved on H2712 carcinomas when the animals were also treated orally by TX or TO. Using the dextran-coated charcoal assay, H2712 cells were found to be negative for classical estrogen receptors. CONCLUSIONS: These results indicate that the anti estrogens, TX and TO, have the ability to suppress the growth of the estrogen receptor negative mammary carcinoma and to amplify target cell lysis by tumor reactive CTL. By this mechanism these drugs enhance host immunity to an estrogen receptor negative tumor. PMID- 9413218 TI - Combination immunotherapy of the P815 murine mastocytoma with killer cells, IL-2 and anti-estrogens. AB - BACKGROUND: Lymphokine activated killer (LAK) cells, cytotoxic T lymphocytes (CTL) and interleukin-2 (IL-2) all are being tested in cancer immunotherapy with modest success. The anti-estrogenic drug, toremifene (T0), was found earlier to amplify cancer immunotherapy with killer cells in mice. Here T0 is examined in combination with CTL and IL-2 in immunotherapy experiments. MATERIALS AND METHODS: The P815 mastocytoma of DBA/2 mice was treated with combinations of CTL, LAK cells and T0 and combinations of CTL, IL-2 and T0 along with control groups. Tumor growth rate and mortality has been recorded. RESULTS: Combined treatment with CTL and LAK cells cured 25% of tumor bearing animals, as did treatment with tamoxifen (TX) or T0 alone. When killer cells were given in conjunction with anti estrogens the cure rate was 75% with both TX and T0. Human recombinant IL-2 significantly inhibited tumor growth but no cures occurred. CTL alone cured 25% of the animals. When CTL and IL-2 treatment was given jointly with T0 75% of tumor bearing animals were cured. CONCLUSIONS: The results indicate that treatment with anti-estrogens increased the efficiency of immunotherapy by killer cells. This was true also when CTL therapy was supplemented with IL-2 treatment. PMID- 9413219 TI - The effect of castanospermine on the metastatic properties of prostate cancer cells. AB - BACKGROUND: Most deaths from prostate cancer result from the metastatic spread of the disease. Castanospermine has been shown to inhibit tumor growth and metastasis in mouse and rat models. We hypothesized that castanospermine might inhibit metastasis in the Dunning model of rat prostate adenocarcinoma by interfering with the metastatic properties of tumor cells. MATERIALS AND METHODS: We examined the cytotoxicity of castanospermine toward the metastatic MAT-LyLu and nonmetastatic AT. 1 cell lines and its effects on cell motility and adhesion to endothelial cells. We assessed castanospermine's effects on in vivo metastasis in Copenhagen rats. RESULTS: Castanospermine was not cytotoxic toward the MAT LyLu and AT. 1 cell lines at concentrations through 10 micrograms/mL, nor did it significantly affect cell motility, adhesion to endothelial cells, or in vivo metastasis. CONCLUSIONS: Within the Dunning model, castanospermine did not appear to significantly affect cell characteristics related to metastatic potential. PMID- 9413220 TI - Relationships between cathepsin-D, pS2 protein and hormonal receptors in breast cancer cytosols: inconsistency with their established prognostic significance. AB - In this study the unexpected findings from the analysis of 278 breast cancer tissue specimens are reported. A surprising strongly positive correlation between an unfavourable and a favourable prognosis with markers cathepsin D and pS2 respectively, was revealed by linear regression analysis (Pearson, Student-T Test). In the relevant literature reviewed only one similar, although indirect, observation was found. On the other hand, a weak relationship between pS2 and ER has emerged using the same method, while the pS2/PgR association remained strong. The latter supports the hypothesis that pS2 positivity is associated with positive PgR and may be a marker of functioning ER, irrespective of ER status. These and other similar findings underline the need for a better understanding of the underlying molecular events as well as the necessity of an effective prognostic evaluation model for breast cancer. PMID- 9413221 TI - 5-fluorouracil (5-FU) and 5,10-methylene tetrahydrofolate (5,10-CH2FH4) as adjuvant therapy in an experimental rodent colon carcinoma model. AB - Eradication of micrometastases is the goal for adjuvant therapy following a radical surgical procedure for cancer. We report an experimental study with 5,10 methylenetetrahydrofolate (5,10-CH2FH4) modulation of 5-fluorouracil (5-FU) cytotoxicity in adjuvant treatment. A colon adenocarcinoma cell suspension was inoculated intrahepatically in a rodent experimental model. Intravenous 5-FU (30 mg/kg) in combination with 5,10-CH2FH4 (15 mg/kg or 30 mg/kg) was administered after 1, 2, 3, 4 and 7 days. 5-FU alone reduced the tumor take to fifty percent compared to one hundred percent tumor take in control animals (p < 0.05), while 5 FU in combination with 5,10-CH2FH4 (regardless of folate-dose) eliminated tumor take (p < 0.0001). This makes 5,10-CH2FH4 a promising agent for modulation of 5 FU cytotoxicity in adjuvant cancer treatment. PMID- 9413222 TI - Mechanism of the cytotoxicity of asterriquinone, a metabolite of Aspergillus terreus. AB - Asterriquinone (ARQ) is an antitumor metabolite of Aspergillus terreus IFO 6123. ARQ is a quinone compound and the mechanism of its antitumor action was compared with that of its analog ARQDMe which was dimethylated at the dihydroxy benzoquinone moiety, and adriamycin (ADR). The cytotoxicity of ARQ against P388 cells was less than that of ADR, and ARQDMe showed hardly any cytotoxicity. ARQ and ADR formed DNA-interstrand cross links (ISC) in P388 cells in a concentration while the ISC index by ARQDMe was very low even at 10 microM. P388 cells treated with ARQ and ADR for 18 hours showed, dose-dependently, nucleosomal ladder formation as well as the appearance of degraded DNA. Difference between the action of ARQ and ADR were observed by flow cytometric cell cycle analysis. ARQ caused the cells to accumulate at the G1 phase of the cell cycle, while use of ADR led to arrest in the G2/M phase. ARQDMe never caused apoptotic changes or cell cycle alterations. These results indicate that ARQ intercalates in the genomic DNA, causes G1 arrest, and leads to apoptotic cell death, this suggests that the dihydroxybenzoquinone moiety of the compound is essential to elicit these actions. PMID- 9413223 TI - Pharmacokinetics of platinum following the administration of cis-(glycolato O,O')[(4R,5R)-4,5-bis(aminomethyl)-2-isopropyl-1, 3-dioxolane]platinum(II) in dogs. AB - The pharmacokinetic study on cis-(glycolato-O,O') [(4R,5R)-4,5-bis(aminomethyl)-2 isopropyl-1,3-dioxolane]-platinum(II) (SKI 2034R, NSC D642489) was performed in beagle dogs. A single dose of 2.0 mg/kg of SKI 2034R was given by i.v. bolus to three beagle dogs. Plasma samples were analyzed for platinum by flameless atomic absorption spectrophotometry. Plasma concentrations of total and ultrafiltrable platinum for SKI 2034R declined in a biexponential fashion. The mean area under the concentration-time curve (AUC0-->infinity) determined for ultrafiltrable platinum derived from SKI 2034R, as an active component, was 2.76 +/- 0.13 micrograms.h/ml (mean +/- S.D.), with an initial half-life of 0.15 +/- 0.09 hour, a terminal half-life of 0.96 +/- 0.12 hour, a total clearance of 12.07 +/- 0.67 ml/min/kg, and a steady-state volume of distribution of 0.82 +/- 0.06 1/kg. PMID- 9413224 TI - Immunolocalization of cyclins D and E and cyclin dependent kinase (cdk) 2 and 4 in human breast carcinoma. AB - We studied the immunolocalization of cyclins D1 and E and their corresponding partner cyclin dependent kinases (cdk), cdk4 and cdk2 in 41 cases of human breast malignancy (21 invasive ductal carcinomas and 19 invasive lobular carcinomas) and examined the correlation of the labeling indexes among these cyclins, cdks, Ki67, estrogen receptor (ER) and progesterone receptor (PR). Cyclin D1 immunoreactivity was observed exclusively in the nuclei of tumor cells in 27/41 (65%) of the cases examined. Immunoreactivity for cyclin E and cdk2 was detected in all the cases and observed in the nuclei of both carcinoma and non-carcinoma cells. cdk4 immunoreactivity was detected in 39/41 (95%) cases and found in carcinoma and non carcinoma cells. In all carcinomas examined, a significant correlation was observed only between Ki67 and cyclin D1 (p = 0.0037). However, when examining only invasive ductal carcinomas, a significant correlation was detected between Ki67 and cyclin D1 (p = 0.0069), Ki67 and cdk2 (p = 0.0043) and cyclin D1 and cdk4 (P = 0.0024). Only cyclin D1 correlated with the pathologic stages of the disease and histological grades of invasive ductal carcinoma. Among these cyclins and cdk, overexpression of cyclin D1 is considered to play an important role in the development of human breast malignancy through abnormal proliferation. No significant correlation was observed between steroid receptor status and any of cyclins and cdks examined. Cyclin D1 and cdk2 expression correlated with cell proliferation (Ki67) and cyclin D1 expression with expression of cdk4 in invasive ductal carcinoma but not invasive lobular carcinoma. Cyclin E expression did not correlate with cell proliferation, cyclin D1 or cdks possibly due to deregulation of its expression. These results also indicate different patterns of cyclin D1, cyclin E, cdk2 and cdk4 expression between invasive ductal and lobular carcinoma of human breast. PMID- 9413225 TI - Lectin immunohistochemistry study of nasal inverted papilloma and associated neoplasms. AB - BACKGROUND: We aimed to investigate the tumor biology of nasal IP and identify the biological characteristics associated with early diagnosis and malignant transformation. MATERIALS AND METHODS: Lectins immunohistochemistry were performed on inverted papilloma (IP), and associated neoplasms. IP synchronized with polyp or squamous cell carcinoma (SCC) and IP metachronized with SCC were analyzed and correlated to clinical information. RESULTS: Canavalia ensiformis (ConA) and Arachis hypogaea (PNA) with neuraminidase pretreatment (NA-PNA) showed similar staining in both the IP and SCC portions of the IP synchronized with SCC. The IP and polyp portions of an IP synchronized polyp had positive NA-PNA staining, while papilloma and polyps alone had negative staining. Strong NA-PNA staining in the IP (transformed to SCC) showed significant differences from IP. CONCLUSION: These biological characteristics define IP as a premalignant neoplasm. NA-PNA staining may be helpful for an early detection of IP. Strong NA PNA staining in IP may predict malignant transformation. PMID- 9413226 TI - Differential expression of growth arrest, DNA damage genes and tumour suppressor gene p53 in naevi and malignant melanomas. AB - Twenty nine benign naevi and 59 melanomas of different Clark level and locations were stained immunohistochemically for the presence of p53 and 3 members of Growth Arrest. DNA Damage inducible family: GADD 34, GADD 45 and GADD 153. All naevi were p53 negative and highly GADD positive. Differences in the expression of all 3 GADD proteins between naevi and melanomas were highly significant (p = 0.0001). The expression of p53 increased and the expression of GADD proteins decreased with the Clark level of melanoma thickness. The survival time of patients correlated negatively with p53 positivity and positively with GADD 45 and 153 expression. Prognostic significance for GADD 34 was not found. Our conclusion is that GADD proteins play an important role in the malignant transformation of naevus to melanoma and GADD 45 and GADD 153 proteins can influence patient survival. PMID- 9413227 TI - The correlation between the expression of p53 protein and DNA ploidy in patients with gastric cancer that has invaded the serosa. AB - To estimate the correlation between the expression of mutated p53 protein and the nuclear DNA ploidy of gastric cancer cells, as well as the influence of these factors on the prognosis of patients with advanced gastric cancer, we selected 161 patients with gastric cancer that had invaded the serosa and who were treated by potentially curative gastrectomy. The expression of p53 protein was detected in 95 (59%) patients and aneuploidy was detected in 70 (43.5%) patients. The percentage of tumors with DNA aneuploidy among the 95 tumors that were p53 positive was 51.6% and this percentage was significantly higher than that of tumors with DNA aneuploidy among the 66 tumors that were p53-negative (31.8%, P = 0.02). The percentage of cells in the G2M phase of the cell cycle was significantly higher in p53-positive tumors than that in p53-negative tumors (P = 0.024). The 161 patients were divided into four groups according to the expression of p53 protein and the DNA ploidy of their tumors. Patients with p53 positive tumors and DNA aneuploidy were allocated to group A (n = 49), patients with p53-positive tumors and DNA diploidy were allocated to group B (n = 46), patients with p53-negative tumors and DNA aneuploidy were allocated to group C (n = 21), and patients with p53-negative tumors and DNA diploidy were allocated to group D (n = 45). The 5-year survival rate of group D was 73.7% and it was significantly higher than that of group A (41.8%, P = 0.007). These observations indicate that mutated p53 protein might accelerate cell proliferation. Moreover, analysis of both p53 status and DNA ploidy of tumors seems to provide very important information for evaluation of the prognosis of patients with advanced gastric cancer. PMID- 9413229 TI - Heart disease and stroke in Canadian women. PMID- 9413228 TI - Evaluation of topoisomerase I catalytic activity as determinant of drug response in human cancer cell lines. AB - The prognostic value of topoisomerase I (Topo I) catalytic activity and expression of the multidrug resistance (MDR) marker P-glycoprotein (Pgp) and multidrug resistance protein (MRP) for in vitro sensitivity to Topo I interactive agents were evaluated. The efficacy of short term (2 h) and long term (24 h) exposures of camptothecin (CPT), two CPT derivatives (SN-22, SN-38) and the indolocarbazole compound NB-506, was determined against human ovarian carcinoma (A2780 and A2780 DX5), human fibrosarcoma (HT1080 and IIT1080/DR4) and human ileocecal carcinoma (HCT-8). For each cell line the Topo I protein levels and catalytic activity were determined and correlated with drug-induced cytotoxicity. In general, the Topo I protein levels correlated with Topo I catalytic activity. Drug-induced cytotoxicity increased significantly with prolongation of the exposure time. With the 2 h exposure, the multidrug resistant A2780 DX5 cell line (Pgp+, MRP-) was moderately resistant to all four drugs compared to its parental cell line. In case of CPT and SN-22 but not for SN-38 and NB-506, this resistance was no longer detectable following 24 h drug exposure. No resistance was detectable for the multidrug resistant HT1080/DR4 (Pgp-, MRP+) cell line when compared to its parental cell line. With short term exposures a strong trend was observed toward increased cytotoxicity with increased Topo I catalytic activity, especially if this correlation was studied between derivative cell lines (A2780 vs. A2780 DX5 and HT1080 vs. HT1080/DR4). This correlation weakened when all 5 cell lines and both exposure conditions were considered. Thus, although overall the correlation between Topo I catalytic activity and sensitivity to Topo I interactive drugs between different cell lines is weak, this correlation may be stronger when comparing derivative cell lines. For CPT and SN-22 but not for SN 38 and NB-506, the moderate resistance levels observed in the Pgp-expressing cell line could be negated by prolongation of exposure duration. MRP expression did not effect drug efficacy. The data demonstrate that the importance of Topo I catalytic activity as single prognostic factor for drug response to Topo I interactive agents is weak and that additional mechanisms affecting drug response have to be taken into consideration. PMID- 9413230 TI - Re: D Waters. Calcium channel blockers: an evidence-based review. 1997;13:757-66. PMID- 9413231 TI - Does Chlamydia pneumoniae cause coronary atherosclerosis and should we all take macrolides? PMID- 9413232 TI - Oxidative stress and cardiovascular disease. AB - OBJECTIVE: To assess the evidence supporting the role of oxidative stress in the pathogenesis of atherosclerosis, endothelial dysfunction, platelet aggregation and heart failure, and to evaluate the status of antioxidant therapy in the clinical management of cardiovascular disease. DATA SOURCES: No attempt was made to provide insight into mechanisms of oxidative stress and the response to antioxidant intervention. Human studies were scrutinized for evidence that oxidative stress was present and whether antioxidant therapy provides clinical benefit. CONCLUSIONS: There is ample evidence that oxidative stress is involved in the pathogenesis of atherosclerosis, endothelial dysfunction, platelet aggregation and heart failure. However, the extrapolation of this evidence to the use of antioxidant therapy in primary or secondary prevention of cardiovascular disease cannot be justified at this time. PMID- 9413233 TI - Systolic function of the univentricular heart: comparison of the Simpson's rule with acoustic quantification. AB - BACKGROUND: Acoustic quantification (AQ) is a new method of obtaining real-time information about systolic ventricular function. This method establishes a ;blood tissue interface' and computes an intraventricular blood volume in real time to derive a beat to beat instantaneous ejection fraction. AQ assessment of systolic function has been reported previously in patients with normal cardiotypes and varying degrees of myocardial dysfunction. OBJECTIVE: To determine the potential utility of AQ in patients with abnormal ventricular morphology, in whom systolic function may be difficult to measure by traditional methods. PATIENTS AND DESIGN: Seventeen children (nine females) ranging in age from five days to 18 years (mean 6.9 years) with univentricular left ventricle heart morphology underwent a prospective and comparative echocardiographic study of ventricular function with the use of AQ and manual planimetry (single plane Simpson's rule). Imaging was done during steady state without sedation. Routine scan planes were performed, followed by repeat scanning of the univentricle from the apical four-chamber view in the AQ mode. Subsequently, manual planimetry using Simpson's rule was performed from an online graphical analysis package to measure systolic and diastolic frames from the conventional replay images. These data were used to calculate ejection fraction using standards previously established. The results were then compared with real-time AQ results. SETTING: Tertiary care referral center. RESULTS: Scan time for the combined standard and AQ imaging averaged 45 mins (range 35 to 65 mins). Measured ejection fraction by AQ and manual planimetry were 44 +/- 11% and 46 +/- 10%, respectively. Statistical analysis by repeated measures ANOVA with Bonferroni/Dunn correction (F = 0.6, df = 1,32, P = 0.44) demonstrated significant agreement between AQ and manual planimetry with an intraclass correlation coefficient of 0.93. Bland-Altman analysis was used to provide a graphic display of the clinical significance of differences in the comparison of the two methods of measurement. CONCLUSIONS: These findings support the use of AQ for continuous online determination of indexes of systolic function for patients with univentricular left ventricle morphology. The variability in the morphology inherently present within this group of patients results in a wider variability of determined ejection fraction. Particular attention must be directed to the technical aspects of image acquisition and AQ application to ensure accuracy. PMID- 9413234 TI - Early diagnosis of acute myocardial infarction by either electrocardiogram or a logistic regression model: portability of a predictive instrument of acute cardiac ischemia to a small rural coronary care unit. AB - OBJECTIVE: To test the ability of a logistic regression model (LRM) that predicts acute cardiac ischemia to make an early diagnosis of acute myocardial infarction (AMI); the ability of the LRM to predict AMI was also compared with the presenting electrocardiogram (ECG). SETTING: A small rural Irish coronary care unit. METHODS: Clinical and ECG data required by the LRM to predict acute coronary ischemia were recorded in 600 consecutive patients admitted with suspected AMI. Estimates of the LRM were ranked into equal deciles in declining probability of acute cardiac ischemia (pACI), and presenting ECGs were placed into one of seven categories. RESULTS: At presentation 50% of AMI patients were in the two LRM deciles with the highest pACI, and 49% of AMI patients had ECGs with greater than 2 mm ST elevation associated with reciprocal changes. ECG categories had a 76% sensitivity for the early diagnosis of AMI and the LRM had an 84% sensitivity. The specificity, accuracy and positive predictive value for the ECG categories were 92%, 84% and 85%, respectively. The specificity, accuracy and positive predictive value of the LRM were 84%, 84% and 75%, respectively. The areas under the receiver operating characteristic curve of the LRM and ECG categories were almost identical (91% and 90%, respectively). CONCLUSION: AMI can be diagnosed early with comparable accuracy either by placing presenting ECGs into one of seven categories, or by the LRM. The best method and 'cut-off' point for the diagnosis of AMI varies according to clinical circumstances. Categorizing ECGs requires more skill in ECG interpretation, but takes less time. The previously reported performances of the LRM were replicated, confirming portability of its use into different clinical settings and patient populations. PMID- 9413235 TI - Midterm results with the Sorin Monostrut heart valve prosthesis. AB - OBJECTIVE: To monitor the hematological and clinical sequelae of a single tilting disc cardiac valve prosthesis. DESIGN: Prospective nonrandomized trial. SETTING: University teaching hospital. PARTICIPANTS: All patients receiving a single mechanical cardiac valve prosthesis were offered the Sorin Monostrut valve if they met the criteria for valve use. Seventy-five per cent of the patients entered were in New York Heart Association (NYHA) functional class III or IV. One hundred and forty-seven patients were subsequently followed at three months and then yearly after valve implantation for seven years. MAIN OUTCOME MEASURES: At one year, preoperative indexes of hemolysis were compared with three-month and one-year postoperative values. Actuarial curves for survival, freedom from cerebrovascular events and explantation were constructed for the seven-year follow-up period. RESULTS: Hemolysis, as measured by lactate dehydrogenase values, commonly occurs preoperatively, remaining significantly elevated three months and one year following valve implantation. Serum haptoglobin was normal preoperatively but was significantly low at one year. Anemia was uncommon and most patients had normal reticulocyte counts at one year. At three years, 81% of patients were in NYHA functional class I. CONCLUSIONS: Midterm results show that this valve is structurally reliable and meets all current requirements for a safe mechanical valve. PMID- 9413236 TI - Acute aortic dissection and ventricular septal defect repair. AB - A 37-year-old male with long-standing heart murmur and ventricular septal defect developed acute chest pain and was diagnosed with an aortic dissection and aortic insufficiency. The ventricular septal defect and aortic dissection were repaired successfully as a combined procedure. At late follow-up (three years), he continued to enjoy an excellent result. PMID- 9413237 TI - Thomas W Smith MD: 1936 to 1997. PMID- 9413238 TI - Atrial fibrillation: extending basic science to clinical frontiers. AB - Basic research in atrial fibrillation is advancing with enormous speed, extending the boundaries from research in intact tissues, to cellular electrophysiology and to molecular biology. Never before has the need been greater for a true 'bench to bedside' approach to research. The basic researchers need to understand the potential clinical relevance of their work so that their efforts may be directed to areas of clinical impact. On the other hand, the clinician needs the aid of the basic researcher to help solve some of the vexing clinical problems. Emphasizing the need for this liaison, this paper discusses problems confronted by the clinician and suggests areas of basic research that may help answer the frustration of the clinician in dealing with patients with atrial fibrillation. PMID- 9413239 TI - Electrical activity in Koch's triangle. AB - The authors have conducted several experimental studies of the cellular electrophysiology of the atrioventricular (AV) node employing the Langendorff blood perfused heart of both dogs and pigs. Two types of experiments are described: experiments showing that cells with electrophysiological characteristics of typical nodal cells can be found outside Koch's triangle; and mapping experiments during the induction of ventricular echo beats in an attempt to delineate the reentrant circuit thought to underlie AV nodal reentry. PMID- 9413240 TI - Implications of ion channel diversity to ventricular repolarization and arrhythmogenesis: insights from high resolution optical mapping. AB - Recent evidence from experiments in isolated cells suggests that substantial ion channel diversity exists throughout the heart. However, due to limitations of conventional electrophysiological recording techniques, the influence of ion channel diversity on ventricular repolarization and arrhythmia vulnerability at the level of the intact heart are poorly understood. High resolution optical mapping with voltage-sensitive dye was used to measure significant heterogeneity in the kinetics of cellular repolarization between cells across the epicardial surface (1 x 1 cm) of the intact guinea-pig heart. Such diversity of repolarization kinetics modulates dispersion of repolarization in a biphasic fashion as premature coupling interval is shortened. Moreover, evidence is provided that modulation of repolarization by a premature stimulus in a coupling interval-dependent manner also modulates arrhythmia vulnerability in parallel, ie, 'modulated dispersion' hypothesis. Therefore, due to diversity of repolarization kinetics between epicardial cells, a premature stimulus serves not only as a 'trigger' of arrhythmias, but also importantly modulates spatial dispersion of repolarization and the arrhythmogenic substrate for reentry. PMID- 9413241 TI - Voltage-sensitive dye recordings of electrophysiological activation in a Langendorff-perfused mouse heart. AB - The pattern of electrophysiological activation of adult mouse ventricles was measured with the use of voltage-sensitive dye methods. Di-4-ANEPPS was used to monitor membrane potential as small changes in fluorescence, which were detected by a state of the art, cooled, charged coupled device camera/image intensifier system. The extremely rapid conduction velocity, coupled with the small size of this preparation, necessitated taking these measurements at room temperature (22 to 23 degrees C). Initial experiments demonstrate that ventricular activation can be identified and its conduction pattern can be monitored reproducibly and with high resolution for extended time periods (10 to 20 mins) during spontaneous activity. PMID- 9413242 TI - Interactions between spontaneously pacing and quiescent but excitable heart cells. AB - The authors previously developed a technique for studying a mathematical model cell with spontaneous activity, namely, a 'real time' simulation of a rabbit sinoatrial node (SAN) model cell that is simultaneously electrically coupled via a 'coupling clamp' circuit to a real, isolated ventricular myocyte. This technique was applied to investigate the effects of coupling conductance (Gc), cell size and modulation of membrane potential by elevated extracellular potassium ion concentrations on the ability of an ectopic focus, represented by the SAN model cell, to successfully drive a ventricular cell. Values of Gc and the relative sizes of the two cells define three possible outcomes: spontaneous pacing of the SAN model cell but not driving of the ventricular cell; cessation of spontaneous pacing; or pacing of the SAN model cell and driving of the ventricular cell. Below a critical size of the SAN model cell, only the first two outcomes are possible. Above this critical size, there is a range of Gc that allows successful operation of the system as an ectopic focus. Elevation of extracellular potassium ion concentrations from 4 to 8 mM increases both the lower and upper boundaries of Gc for this range. Elevation of extracellular potassium ion concentrations, commonly observed in myocardial ischemia, may affect either inhibition or release of inhibition of an ectopic focus. PMID- 9413243 TI - Reversible inhibition of gap junctional communication elicited by several classes of lipophilic compounds in cultured rat cardiomyocytes. AB - BACKGROUND: Electrical coupling between cardiac muscle cells is mediated by specialized sites of plasma membrane termed 'gap junctions', which consist of clusters of transmembrane channels that directly link the cytoplasmic compartments of neighbouring cells and allow direct transfer of small ions and molecules. These structures provide low resistance electrical pathways between cardiac cells, necessary for rapid impulse propagation and, thus, coordinate contraction of the myocardium. OBJECTIVE: To investigate the effects of derivatives of sex steroid hormones and of some of their antagonists on junctional communication in pairs of cultured ventricular myocytes of neonatal rats. MAIN RESULTS: Short term (15 min) exposures to some of these lipophilic compounds led, in a concentration range 1 to 22 microM, to a reversible inhibition of cell to cell communication. None of these uncoupling treatments altered the cytosolic calcium concentration, examined by means of a fluorescence indicator. The uncoupling effect of sex hormones persisted in the presence of blockers of their respective nuclear receptors (eg, cyproterone acetate for testosterone and tamoxifen for 17-beta-estradiol). Some of these blockers (tamoxifen, clomiphene) were able to impair gap junctional communication, whereas others (nafoxidine and cyproterone acetate) had no effect. None of protein kinase C, cAMP-dependent protein kinase and protein tyrosine kinase pathways seemed to be involved in these effects. CONCLUSIONS: Several lipophilic compounds able to hinder cell to cell communication have also been seen to affect voltage-activated or ligand-activated ionic channels. Lipophilic molecules with an appropriate molecular skeleton could insert into the membrane, with resulting destabilization and unspecific closure of membrane channels. PMID- 9413244 TI - Effects of 8-oxoberberine on sodium current in rat ventricular and human atrial myocytes. AB - 8-Oxoberberine (JKL1073A), a derivative of berberine, has been reported to exert positive inotropic action and antiarrhythmic activity, much like a class III antiarrhythmic agent. In addition to prolongation of cardiac action potential duration, JKL1073A also decreases the maximal rate of action potential upstroke. To characterize the mode of inhibition of action potential upstroke, the effects of JKL1073A on the sodium inward current (INa) of rat and human cardiac myocytes were examined by voltage clamp in a whole cell configuration. In rat ventricular myocytes, JKL1073A and quinidine inhibited INa with an average 50% inhibitory concentration (IC50) of 3.3 +/- 0.2 microM (n = 9) and 2.1 +/- 0.1 microM (n = 6), respectively. Voltage-dependent steady state inactivation curves of INa were shifted slightly by 10 microM JKL1073A but more significantly by 3 microM quinidine to negative potential. Though steady state inactivation of INa was not significantly changed by 3 microM JKL1073A, the time constant of INa recovery from inactivation state was partially prolonged from 51.7 +/- 18.5 ms to 74.1 +/- 23.8 ms. Quinidine (3 microM) prolonged the time course of INa recovery from inactivation with an associated increase of slow recovery component. Inhibition of INa by JKL1073A and quinidine was increased when cells were stimulated at higher frequency. The maximal INa obtained in cells held at -140 mV was significantly decreased by 10 microM quinidine to 45.3 +/- 2.5% of control but only partially suppressed by 10 microM JKL1073A to 85.9 +/- 8.6% of control. In human atrial myocytes, JKL1073A and quinidine suppressed INa with an average IC50 of 2.4 +/- 0.6 microM and 1.4 +/- 0.3 microM, respectively. Both JKL1073A and quinidine increased the time constant of INa recovery from inactivation. In conclusion, JKL1073A at concentrations of 3 microM and 10 microM may inhibit INa mainly by binding to open and inactivated channels. Quinidine 3 microM may inhibit INa by binding to resting, open and inactivated channels. Inhibition of INa by JKL1073A may explain the inhibition of the action potential upstroke and contribute partially to its antiarrhythmic activity. PMID- 9413245 TI - Electrophysiological analyses of threshold conditions and rate-dependent failure of excitation in single myocytes from rabbit ventricle. AB - The changes in transmembrane ionic currents that underlie normal excitability and rate-dependent failure were studied in single cells from rabbit ventricle by using whole cell voltage clamp methods. When trains of brief (1 to 2 ms) stimuli are applied at strengths very close to the threshold for excitation, a number of different patterns of action potential entrainment and failure are observed. In an individual cell, a characteristic pattern of entrainment or failure can be maintained for a relatively long time, allowing both a detailed description and a quantitative investigation of the ionic basis for this phenomenon. Three hypotheses for rate-dependent failure of excitation in rabbit ventricle were examined. The first is that following relatively high rates of stimulation, the intracellular calcium ion concentration increases and, secondarily, a background inwardly rectifying potassium ion current (IK1) decreases, thereby lowering the excitation threshold. The second hypothesis is that residual activation of the delayed rectifier potassium ion current (IK) causes the stimulus to become subthreshold as the rate of stimulation increases. The third hypothesis is that small changes in the time and voltage dependence of the inactivation and reactivation of the sodium ion current (INa) result in less net inward ion current for a given waveform of depolarization, and the cell therefore becomes inexcitable (eg, to every second stimulus). The calcium ion hypothesis was tested by buffering changes in intracellular calcium ion concentrations with BAPTA. The results strongly suggest that changes in intracellular calcium ion concentrations do not contribute significantly to the observed patterns of failure of excitation. The delayed rectifier hypothesis was evaluated using the class III antiarrhythmic drug dofetilide, which selectively blocks a large fraction of the IK current in rabbit ventricle. Dofetilide slightly decreased the stimulus threshold, suggesting that residual activation of the rapidly activated outward conductance of potassium ions is an important variable under some conditions. The INa hypothesis was tested by altering the size of INa with tetrodotoxin (TTX). After TTX application, the threshold for excitation increased significantly, and rate-dependent failure and entrainment were no longer observed until the stimulus strength was increased. These results show that INa is an essential variable underlying normal excitation and entrainment in rabbit ventricle. This is plausible because INa is 20 to 50 times larger than either the maximum outward current due to IK1 or the fully activated IK in this tissue. PMID- 9413246 TI - Gastric Cancer. PMID- 9413247 TI - Thrombolysis for acute ischemic stroke. PMID- 9413248 TI - Geriatrics photo quiz. Bell's palsy (seventh nerve palsy). PMID- 9413249 TI - Red, cup-shaped nodule. PMID- 9413250 TI - Guideline for initial evaluation of the patient with memory loss. AB - A practical and useful guideline for early identification of Alzheimer's disease and related dementias has been developed by the Agency for Health Care Policy and Research. By using the initial evaluation process described in this guideline, primary care physicians can identify and treat many "reversible" causes of memory problems, such as depression, delirium, and co-morbid medical conditions. The treatable causes of dementia may be structural, metabolic, toxic, infectious, nutritional, psychiatric, or drug-related. Initial evaluation includes patient history, physical examination, mental status testing, functional assessment, and some laboratory tests. Information from an informed family member can be very helpful in this process. PMID- 9413251 TI - Heart protection: controlling risk factors for cardiovascular disease. AB - Cardiovascular disease is the leading cause of illness and death in the United States. Clinical data continue to support primary prevention through the aggressive treatment of well-defined cardiovascular risk factors. Three risk factors that can be modified to lower the risk of cardiovascular disease and death are hypercholesterolemia, hypertension, and cigarette smoking. Even patients with asymptomatic cardiovascular disease have been shown to benefit from aggressive cholesterol-lowering therapy. New JNC-VI guidelines for managing hypertensive disease recommend that treatment decisions be based on level of blood pressure plus presence or absence of target organ damage or other risk factors. The risk of myocardial infarction in former smokers approaches that of nonsmokers after 3 years. PMID- 9413253 TI - Lack of effect of Miller Fisher sera/plasmas on transmitter release from PC12 cells. AB - IgG antibodies to GQ1b ganglioside are found in > 90% of patients with the Miller Fisher Syndrome (MFS). MFS sera or IgG preparations have marked effects on neurotransmitter release at the neuromuscular junction, but their mode(s) of action remain unclear. To establish a cell-based system for investigating the mechanism of action of MFS serum preparations, we looked at neurotransmitter release from three cell lines. We failed to demonstrate substantial 14C acetylcholine release from two motor-neuronal cell lines, VSC4.1 and NSC19, and therefore studied 3H-noradrenaline release from NGF-differentiated PC12 cells, a neural-crest derived catecholaminergic cell line. K(+)-induced release was inhibited by botulinum toxin and basal release was enhanced by alpha-latrotoxin, resembling that at the neuromuscular junction, although K(+)-induced release was dependent on L-type rather than P/Q-type calcium channels. The cells expressed polysialylated gangliosides on the cell surface. Incubation in heat-inactivated or untreated MFS preparations did not, however, affect basal or K(+)-induced release. Thus the PC12 cells do not appear to be sensitive to the effects of serum antibodies from MFS patients. PMID- 9413254 TI - Suppression of A beta-induced monocyte neurotoxicity by antiinflammatory compounds. AB - Previous work from this laboratory has demonstrated that prior exposure of peripheral blood monocytes (PBM) to aggregated beta-amyloid peptide (A beta), the major protein comprising the amyloid plaques characteristically present in the brain of Alzheimer disease (AD)-afflicted individuals, activates these cells to a neurotoxic state when co-cultured with brain tissue. In this report we extend these findings to further show that such A beta-induced PBM neurotoxicity can be inhibited by three differentially-acting antiinflammatory drugs, indomethacin, dexamethasone, and colchicine, which are typically used clinically to treat peripheral inflammatory disease. In addition, evidence is presented that these toxic effects are initiated, in large part, by soluble factors released from A beta-stimulated PBM. Our results suggest a rationale for antiinflammatory therapy in the treatment of AD. PMID- 9413255 TI - TCRV and TCRJ gene usage in MBP responding T cells from (B10.PL x PL/J)F1 mice is biased towards that of B10.PL mice. AB - We analyzed myelin basic protein (MBP) specific T cell hybridoma clones from (B10.PL x PL/J)F1 mice. MBP-reacting T cell hybridomas from F1 mice preferentially expressed B10.PL TcraV2.3 (53%) and B10.PL TcraV4.2 (13%) with minor expression of TcraV4.4 (13%) gene segments. A dominant expression of TcrbV8.2 (73%) accompanying with TcrbV8.1 (20%) and TcrbV13 (7%) gene segments have been identified in these MBP-reacting T cell hybridomas from F1 mice. There was less restrictive but non-random usage of the TcraJ and TcrbJ gene segments. Overall, the MBP-reacting T cell hybridomas from (B10.PL x PL/J)F1 mice were dominated by the MBP-reacting T cell pattern seen in B10.PL mice. PMID- 9413256 TI - Human IgM paraproteins demonstrate shared reactivity between Campylobacter jejuni lipopolysaccharides and human peripheral nerve disialylated gangliosides. AB - IgM paraproteins from patients with CANOMAD (chronic ataxic neuropathy, ophthalmoplegia, M-protein, agglutination, anti-disialosyl antibodies) react with NeuAc(alpha 2-8)NeuAc epitopes on a wide range of gangliosides including GQ1b, GT1a, GD1b and GD3. The tissue distribution of reactive antigens in human peripheral nerve has not been addressed in detail. In addition, the origin of these antibodies is unknown. Here we report that purified anti-disialosyl paraproteins from two affected patients bind a wide array of human peripheral nerve structures including dorsal root ganglia, dorsal and ventral root axons, femoral and oculomotor nerves. We also show that these paraproteins bind lipopolysaccharides of Campylobacter jejuni isolates from 3/3 cases of Miller Fisher syndrome, and to a less frequent extent, from cases of Guillain-Barre syndrome and enteritis controls. In conjunction with our previous studies, these data provide a possible causal link between the origin and pathogenic effects of anti-disialosyl antibodies in human paraproteinaemic neuropathy. PMID- 9413257 TI - Genetic analysis of inflammation, cytokine mRNA expression and disease course of relapsing experimental autoimmune encephalomyelitis in DA rats. AB - Genetic analysis of experimental autoimmune encephalomyelitis (EAE) can provide clues to the etiology of multiple sclerosis (MS). Identifying the susceptibility genes of DA rats may be particularly rewarding since they are prone to develop a remarkably MS-like chronic and demyelinating disease. As a first step in this direction, we investigated the role of DA genes within and outside the major histocompatibility complex (MHC) for susceptibility to severe protracted and relapsing EAE (SPR-EAE). This form of EAE developed in DA rats but not in LEW. ACI and BN rats after immunization with syngeneic spinal cord and complete Freund's adjuvant. Studies of crosses between DA and BN rats revealed that non MHC genes determine susceptibility to SPR-EAE. A role for MHC-genes was also established using MHC-congenic rat strains, in which the DA MHC haplotype (av1) associated with relapsing EAE. Again, non-MHC genes were decisive since a high incidence of SPR-EAE only occurred in rats with DA non-MHC genes. Analysis of cytokine mRNA expression and infiltrating cells in the spinal cords of congenic strains revealed that the av1 haplotype associated with a high CD4/CD8 ratio and expression of mRNA for interferon-gamma (IFN-gamma), but not for transforming growth factor-beta (TGF-beta) or interleukin-10 (IL-10). In contrast, the other MHC haplotypes (h, l, u) associated with low CD4/CD8 ratios and mRNA expression for TGF-beta and IL-10, but not for IFN-gamma. DA non-MHC genes determined the intensity of inflammation since the number of cells expressing MHC class II, CD4 and interleukin-2 receptor (IL-2R) was higher in DA rats than in LEW.1AV1 and PVG.1AV1 rats which also carry the av1 haplotype. We conclude that the MHC haplotype of DA rats favors a prolonged proinflammatory autoimmune response associated with relapses, while the DA background intensifies inflammation correlating with a high incidence of relapsing disease. PMID- 9413258 TI - Chemokine-enhanced migration of human peripheral blood mononuclear cells is antagonized by interferon beta-1b through an effect on matrix metalloproteinase 9. AB - The increased migration of peripheral blood mononuclear cells (PBMNCs) across a fibronectin (FN) matrix in response to the chemokines RANTES, MIP-1 alpha and MCP 1 is antagonized by interferon-beta-1b (IFN beta-1b). MCP-1 treatment of PBMNCs elevates their mRNA level and secretion of a matrix degrading enzyme, matrix metalloproteinase (MMP)-9, which is abrogated by IFN beta-1b. The clinical benefits of IFN beta-1b treatment in multiple sclerosis patients may in part be a result of this drug's ability to decrease the migration of PBMNCs in spite of a chemotactic gradient. Furthermore, the elevation of MMP-9 production by PBMNCs may be an important mechanism of action of chemokines. PMID- 9413259 TI - Human CD8+ TCR-alpha beta(+) and TCR-gamma delta(+) cells modulate autologous autoreactive neuroantigen-specific CD4+ T-cells by different mechanisms. AB - To investigate the regulatory interactions among autologous T-cells during the course of multiple sclerosis (MS), proteolipid protein peptide-specific CD4+ T cell clones (TCCs) were irradiated and used as immunogens to stimulate purified populations of autologous CD8+ TCR-alpha beta+ and TCR-gamma delta+ T-cells isolated from the peripheral blood of MS patients, patients with other non inflammatory neurological diseases, and healthy blood donors. The resulting blasts were expanded in the presence of hIL-2 and then cloned by limiting dilution. Two different groups of CD8+ TCCs were revealed. A first group of CD8+ TCCs recognized autologous CD4+ T-cells based in their TCRV beta structures (anti idiotypic responsiveness). A second group of CD8+ TCCs recognized Ag activated autologous CD4+ TCCs irrespective of their Ag specificity or TCRV beta expression (anti-ergotypic responsiveness). Both groups showed MHC class I restricted cytotoxicity against CD4+ T-cells and were able to secrete IFN-gamma, TNF alpha/beta and TGF-beta. TCR-gamma delta+ TCCs isolated in response to stimulation with autologous peptide-specific CD4+ TCCs showed only anti-ergotypic cytotoxicity, which was not inhibited by anti-MHC class Ia monoclonal antibodies. Moreover, they were able to secrete IFN-gamma and TNF alpha/beta, but not TGF beta. These data demonstrate that regulatory mechanisms among human autologous T cells can be mediated by cytolytic interactions or by the release of specific cytokines. Furthermore, they provide evidence that CD8+ TCR-alpha beta+ and TCR gamma delta+ cells differ in their patterns of recognition and in their abilities to modulate the immune response mediated by autologous autoreactive CD4+ T-cells. PMID- 9413260 TI - Nasal administration of myelin basic protein prevents relapsing experimental autoimmune encephalomyelitis in DA rats by activating regulatory cells expressing IL-4 and TGF-beta mRNA. AB - This study explores nasal administration of myelin basic protein (MBP) as a potential means of inducing tolerance to relapsing experimental autoimmune encephalomyelitis (PR-EAE), an experimental multiple sclerosis (MS) model that was induced in DA rats by immunization with rat spinal cord homogenate and incomplete Freund's adjuvant. DA rats received a total dosage of 0, 6, 60, 600 micrograms/rat of bovine MBP on ten consecutive days prior to immunization. EAE with typical course was observed in control rats receiving only PBS nasally, and in rats receiving 6 micrograms/rat of MBP. Rats receiving 60 micrograms/rat of MBP developed acute EAE but no relapse during 60 days of observation post immunization (p.i.). Only one of eight rats receiving 600 micrograms/rat of MBP developed slight, transient EAE. This protection was confirmed at the histology level and was associated with decreased levels of MBP-reactive IFN-gamma secreting Th1-like spleen cells on day 13 and 60 p.i. Rats receiving 60 and 600 micrograms/rat of MBP showed decreased serum anti-MBP IgG2b antibody levels on day 60 p.i., and rats receiving 600 micrograms/rat of MBP had marginally increased anti-MBP IgG1 antibody levels in serum compared to control EAE rats. Cytokine mRNA profiles in central nervous system (CNS) and spleen mononuclear cells were evaluated. Dose-dependent reduction of TNF-alpha mRNA expression were observed both in CNS and in splenocytes. Increased IL-4 and TGF-beta mRNA expression were observed in CNS of low (6 micrograms/rat) and median (60 micrograms/rat) dose of MBP tolerized rats and in splenocytes of rats tolerized with 600 micrograms/rat of MBP. We conclude that nasal administration of MBP in DA rat prevents EAE induced by immunization with whole rat spinal cord homogenate that, besides MBP, contains multiple antigenic myelin proteins. A mechanism involving MBP-reactive regulatory cells expressing IL-4 and TGF-beta mRNA acts as part in the induction of this tolerance. PMID- 9413262 TI - Effect of beta-endorphin on cell growth and cell death in human peripheral blood lymphocytes. AB - Beta-endorphin (beta-end) was investigated for its ability to influence sequential metabolic events that accompany the movements of T-lymphocytes into the cell cycle. When cultured lymphocytes are exposed to this endogenous opioid peptide an increase in polyamine transport across cell membrane is observed. This membrane modification is an early cell cycle event, whose enhancement leads to the intracellular polyamine accumulation. It is shown that beta-end is able to enhance spermidine transport and that the exposition of cells to this peptide is perceived as an apoptotic signal. The possible relationship between induction of apoptotic death and enhancement of polyamine uptake is discussed. PMID- 9413261 TI - Cytokine secretion and nitric oxide production by mononuclear cells of patients with multiple sclerosis. AB - Several experimental findings suggest a potential role of excessive nitric oxide (NO) production by macrophages, microglia and astrocytes in the pathogenesis of demyelinating lesions in MS. We assessed the production of nitrites by peripheral blood mononuclear cells (PBMCs) of 15 MS patients (10 F and 5 M) with the R-R form (EDSS: 1-3.0) and in 15 age-matched control subjects. 9 out of the 15 MS patients showed active lesions in MRI at the time of examination. 7 patients were also monitored at the onset, during and following a clinical relapse. Secretion of cytokines by PBMCs was assessed at the basal time and after 24 h of incubation with lipopolysaccharide (LPS). The production of nitrites in the supernatants of PBMCs stimulated and not stimulated with lipopolysaccharide was evaluated. The secretion of IL1 beta, IFN-gamma, TNF-alpha, IL-6 IL-10 and TGF-beta by PBMCs was detected using ELISA methods. The production of NO, both basal and stimulated, was significantly higher in the patients with active lesions than in those without active lesions (p < 0.01). No significant difference was evident between the basal and LPS-stimulated production of NO between control subjects and MS patients without active lesions. During relapses there was a significant increase in NO production by PBMCs compared to the clinical stable stage of the disease (p < 0.0001). This increase was significantly greater in the early stage of relapse than in the late stage (p < 0.04). A decline of NO levels was observed during recovery. Steroid treatment induced a significant decrease in the PBMC NO production of MS patients during exacerbations (p < 0.01). The levels of IL-1 beta, IFN-gamma and TNF-alpha are significantly higher in the supernatants of the PBMCs which produced greater amounts of NO (p < 0.02, p < 0.03, p < 0.01, respectively). On the other hand, NO levels were negatively related to IL-10 and TGF-beta production (R = -75, p < 0.0001 and R = -0.79, p < 0.0001, respectively). The increase production of NO by peripheral blood mononuclear cells demonstrated in our study to be associated with increased production of proinflammatory cytokines could therefore be considered to be a marker of mononuclear cell activation in the peripheral blood of MS patients and, indirectly, of disease activity. Its increased secretion during T cell and monocyte homing in the CNF could contribute to the damage to the blood-brain barrier and the subsequent cytokine-mediated cytotoxic effect to myelin and oligodendrocytes in the white matter of MS patients. PMID- 9413263 TI - Detectable concentrations of Fas ligand in cerebrospinal fluid after severe head injury. AB - When the cell surface molecule Fas is triggered by its agonist Fas ligand the result is apoptosis of these cells and tissue destruction. To elucidate the pathophysiological relevance of Fas ligand in patients with cerebral oedema caused by trauma, we examined its concentrations in cerebrospinal fluid in 18 patients using specific ELISA. Serum and cerebrospinal fluid from healthy people and injured patients without head trauma did not contain detectable Fas ligand. In contrast, cerebrospinal fluid from patients with severe brain injury contained high concentrations of Fas ligand without detectable concentrations in serum. Soluble Fas ligand concentrations in cerebrospinal fluid correlated significantly with severity of brain injury. The Fas-Fas ligand-system may have a pivotal role in causing oedema and local tissue destruction in the brain after severe head injury. PMID- 9413265 TI - Morphine alters macrophage and lymphocyte populations in the spleen and peritoneal cavity. AB - We have previously shown that subcutaneous implantation of a 75 mg morphine pellet results in suppression of the ability of murine splenocytes to mount an antibody response to sheep red blood cells, due in part to a reduction of macrophage function. The present studies used flow cytometry to examine whether the decrement in macrophage function in the spleens of morphine-treated mice results from a reduction in macrophage numbers. Parallel analysis was carried out on non-elicited peritoneal cells. In the spleen, morphine resulted in a reduction in the relative proportion of macrophages and B-cells, with a concomitant increase in the proportion of T-cells. Alteration in the ratio of CD4+ to CD8+ T cells was not observed. In contrast, in the peritoneal cavity, morphine increased the number of macrophages and reduced the number of B-cells. Naltrexone blocked all of the changes in cellular composition. These results support the conclusion that an important mechanism in the immunosuppression seen in the spleens of mice implanted with morphine pellets is a differential reduction in the number of macrophages and B-cells as compared with T-cells. Further, these studies show that subsets of cells of the immune system are differentially affected by morphine in different anatomical compartments. PMID- 9413264 TI - Rabbit IgG distribution in skin, spinal cord and DRG following systemic injection in rat. AB - In order to determine the distribution of antibodies such as anti-NGF following systemic injection in neonates, immunocytochemical techniques were used to examine the localization of rabbit IgG in rat skin, DRG, and spinal cord after treatments with normal rabbit serum or purified rabbit IgG. Daily subcutaneous injections beginning on postnatal day 2 or on day 15 were given for three days. On the fourth day the animals were sacrificed and tissues were processed for rabbit IgG-IR. In the dorsal and ventral spinal cord, staining intensities suggest a substantial increase in the blood-brain barrier during the first two weeks after birth. Staining intensity in the epidermis of the glabrous skin from the hindpaw was substantially lower than in the adjacent dermis. In addition, IgG infrequently accumulated intracellularly in intensely stained patches in the epidermis. IgG was also able to reach relatively high intracellular concentrations in a small number of sensory neurons. The IgG staining pattern in the skin was similar when anti-NGF itself was administered to the animals. The results are discussed in the context of the effects of anti-NGF on the development of nociceptive afferents. PMID- 9413266 TI - Circulating fragments of myelin-associated alpha 6 beta 4 integrin in Guillain Barre syndrome. AB - Guillain-Barre-Strohl syndrome (GBS) is an acute peripheral neuropathy causing reversible myelin damage. alpha 6 beta 4 is a laminin receptor of Schwann cells and myelin. Along with myelin breakdown, alpha 6 beta 4 immunoreactivity might be detected in patients' sera and provide a marker for monitoring GBS course. MAbs to beta 4 and alpha 6 were used in an ELISA test to detect protein in GBS serum samples as in normal individuals. In 66% GBS patients, alpha 6 beta 4 immunoreactivity was detected while controls were negative. The level of beta 4 was followed in different patients and found to fluctuate, always being positive in at least one sample. Treatment lowered immunoreactivity in two beta 4-positive GBS sera. Then, circulating alpha 6 beta 4 fragments represent a novel marker of extensive peripheral myelin damage and may be used to validate clinical diagnosis of GBS, evaluate its course and activity. PMID- 9413267 TI - Evidence for the production of peroxynitrite in inflammatory CNS demyelination. AB - Peroxynitrite, which is generated by the reaction of nitric oxide (NO) with superoxide, is a strong oxidant that can damage subcellular organelles, membranes and enzymes through its actions on proteins, lipids, and DNA, including the nitration of tyrosine residues of proteins. Detection of nitrotyrosine (NT) serves as a biochemical marker of peroxynitrite-induced damage. In the present studies, NT was detected by immunohistochemistry in CNS tissues from mice with acute experimental autoimmune encephalomyelitis (EAE). NT immunoreactivity was displayed by many mononuclear inflammatory cells, including CD4+ cells. It was also observed in astrocytes near EAE lesions. Immunostaining for the inducible isoform of NO synthase (iNOS) was also observed, particularly during acute EAE. These data strongly suggest that peroxynitrite formation is a major consequence of NO produced via iNOS, and implicate this powerful oxidant in the pathogenesis of EAE. PMID- 9413268 TI - Highly purified oligo-His tagged human recombinant alpha(1)-AChR is immunogenic in vivo and suitable for T cell stimulation in vitro in experimental and human myasthenia gravis. AB - Using recombinantly expressed proteins for selection of antigen-specific T cell lines carries a high risk of selecting T cells specific for contaminating proteins. This risk is especially high for very hydrophobic proteins which are notoriously difficult to purify, such as the integral membrane protein acetylcholine receptor (AChR). We prepared a highly purified recombinant AChR by adding an oligo-histidine affinity-tag to the human alpha(1)-AChR and expressing it in E. coli. This allowed purification by Ni-NTA chromatography and subsequent electroelution from preparative SDS gel as purification steps, resulting in complete purity as assessed by silver stain on SDS-PAGE. This protein preparation induced fatal experimental allergic myasthenia gravis in Lewis rats. Furthermore, the protein could be used to select T cell lines from immunized Lewis rats and patients with myasthenia gravis. However, even with this highly purified protein, one of 8 Lewis rat T cell lines and 3 of 7 human T cell lines cross-reacted to E. coli control proteins. The results show that oligo-histidine tagged, highly purified human alpha(1)-AChR is highly immunogenic in vivo and in vitro. PMID- 9413269 TI - Cytokine mRNA expression in CSF and peripheral blood mononuclear cells in multiple sclerosis: detection by RT-PCR without in vitro stimulation. AB - We amplified the mRNA for cytokines in peripheral blood mononuclear cells (PBMC) and cerebrospinal fluid (CSF) cells from 18 multiple sclerosis (MS) patients and 21 other neurological patients, using reverse transcription polymerase chain reaction (RT-PCR). A radioactive hybridization of the amplified DNA allowed quantitation of mRNA levels. Expression of tumor necrosis factor (TNF)-alpha, interferon (IFN)-gamma and interleukin (IL)-10 mRNA was elevated in CSF cells of MS patients. IFN-gamma and IL-10 mRNA levels were higher in MS patients than in other inflammatory neurological diseases. In many MS patients, both proinflammatory and immunoregulatory cytokine messages were detected in the CSF compartment. Such immune reactivity in CSF, as opposed to the peripheral compartment, did not correlate with the clinical activity of the disease. PMID- 9413270 TI - Regulation of immune functions in rat splenocytes after acute and chronic in vivo treatment with CP-55,940, a synthetic cannabinoid compound. AB - Changes in mitogen-induced splenocyte proliferation and NK activity were evaluated after acute (1 h) and chronic (6 d) in vivo treatment of rats with the synthetic cannabinoid compound CP-55,940. At a dose of 0.4 mg/kg i.p. it significantly inhibited the splenocyte proliferative response to PHA and NK activity but half this dose (0.2 mg/kg) had no effect on immune responses. Pretreatment of rats with the cannabinoid receptor CB1 antagonist SR141716A did not antagonize the CP-55,940-induced immunosuppression, excluding the activation of this receptor subtype in the mediation of this effect. When immune function studies were done on rats tolerant to CP-55,940-induced analgesia, full tolerance also developed for the inhibition of splenocyte proliferation and NK activity. The data provided indicate that CB1 cannabinoid receptors are not involved in mediating the acute and chronic effects of cannabinoids on the immune system and suggest a possible implication of CB2 receptor although other modalities of CP 55,940 action can not be ruled out. PMID- 9413271 TI - Association of autonomic nervous hyperreflexia and systemic inflammation in patients with Crohn's disease and ulcerative colitis. AB - The autonomic nervous system modulates gastrointestinal motility, secretion and mucosal immunity. Its dysfunction may be of pathogenetic importance in inflammatory bowel disease (IBD). This study aimed at investigating the autonomic nervous function in patients with IBD. Forty-seven patients with IBD, 28 with Crohn's disease (CD) and 19 with ulcerative colitis (UC), were investigated by means of 5 cardiovascular and 2 pupillary standardized autonomic nervous function tests. In CD and UC, cardiovascular autonomic neuropathy was very rare (0%, 5%), whereas pupillary autonomic neuropathy was more prevalent (21%, 21%). In contrast to autonomic neuropathy, overall cardiovascular (CD: 29%, UC: 26%) and pupillary autonomic hyperreflexia (46%, 37%) were found more often. Patients with CD and UC demonstrated elevated percentiles in the respiratory sinus arrhythmia test as compared to controls (RSA: 82.3 +/- 3.9%, 80.0 +/- 5.9%, controls: 50.0% +/- 1.5%, p < 0.0001). CD patients with, as compared to patients without, RSA hyperreflexia had significantly higher CDAIs (p < 0.001), increased erythrocyte sedimentation rates (p < 0.005) and more often extraintestinal disease manifestations (p < 0.001). UC patients with, as compared to patients without, pupillary latency time hyperreflexia had lower hemoglobin (p < 0.05), lower albumin (p < 0.01) and increased erythrocyte sedimentation rates (p < 0.05). Autonomic hyperreflexia was significantly associated with more severe inflammation and systemic disease in IBD. Hyperreflexia may be a response to inflammation or a pathogenetic element that drives mucosal inflammation. PMID- 9413272 TI - Nicotine induces leukocyte rolling and adhesion in the cerebral microcirculation of the mouse. AB - Nicotine and several related metabolites were examined for their ability to induce leukocyte rolling and adhesion in the cerebral microcirculation of the mouse. A cranial window was surgically prepared for the visualization of the pial microcirculation using an intra-vital microscopy imaging system. Using this technique rhodamine-labeled leukocytes could be visualized and video-recorded as they traveled within the microvessels, and the quantitation of their rolling and adhesion along the pial venule walls was assessed during an off-line video playback analysis. Nicotine was found to produce significant dose-related increases in leukocyte rolling and adhesion. Cotinine, a major nicotine metabolite, did not induce the same degree of leukocyte rolling and adhesion. Mecamylamine, a nicotine antagonist, was found to inhibit the nicotine-induced leukocyte rolling and adhesion. Anti-P-selectin antibody blocked nicotine-induced leukocyte rolling, while anti-CD18 antibody effectively inhibited leukocyte adhesion, but not rolling in similar experiments. Nicotine-induced leukocyte rolling and adhesion were also inhibited by superoxide dismutase and catalase. These data suggest that nicotine, the principle pharmacological agent in cigarette smoke and related tobacco products, acts via a ganglionic-type nicotinic receptor to enhance leukocyte rolling via P-selectin and reactive oxygen radical-dependent mechanisms in cerebral microcirculation of the mouse. PMID- 9413273 TI - Spread of HSV-1 to the suprachiasmatic nuclei and retina in T cell depleted BALB/c mice. AB - Following uniocular anterior chamber inoculation of the KOS strain of HSV-1 in euthymic BALB/c mice, virus spreads from the injected eye to the brain, and from the brain to the optic nerve and retina of the uninjected eye by day 7 post inoculation (p.i.), but the optic nerve and retina of the injected eye are not infected with virus. Infection of the optic nerve and retina of the injected eye is observed only in athymic mice or in mice depleted of both CD4+ and CD8+ T cells. To determine the role of T cells in virus spread, adult female BALB/c mice were thymectomized and T cell depleted. Mice were co-injected with the KOS strain of HSV-1 and RH116, a thymidine kinase-negative mutant of KOS containing the Escherichia coli lac Z gene. Animals were sacrificed on days 3-7 p.i., and the eyes and brains were examined for blue-stained, virus-infected cells. A difference in the timing of virus infection was observed in the area of the suprachiasmatic nuclei only in mice depleted of both CD4+ and CD8+ T cells, and in this group, the contralateral suprachiasmatic nucleus was infected two days earlier. Since one route by which virus could infect the retina of the injected eye is via connections of the contralateral suprachiasmatic nucleus to the ipsilateral optic nerve, these findings suggest that (a) retinitis observed in the injected eyes of mice depleted of both CD4+ and CD8+ T cells results from virus infection of the contralateral suprachiasmatic nucleus followed by spread of virus to the ipsilateral optic nerve and retina and (b) early HSV-1 infection of the contralateral suprachiasmatic nucleus is prevented by a T cell dependent mechanism. PMID- 9413274 TI - Combined effect of HLA-DRB1*1501 and interleukin-1 receptor antagonist gene allele 2 in susceptibility to relapsing/remitting multiple sclerosis. AB - Susceptibility to multiple sclerosis (MS) is associated with HLA-DRB1*1501. Many reports have suggested associations with other loci but these results remain unconfirmed. We studied the IL-1 receptor antagonist (IL-1ra) gene polymorphism and the HLA-DR and DQ allele frequencies by DNA-based methods in both the primary chronic progressive form (PP MS) and the relapsing/remitting form (R/R MS). The frequency of DRB1*1501 and IL-1ra allele 2 were significantly higher in R/R MS. Association was more marked in the female sex and in patients with benign forms of R/R MS. On the other hand DR4 subtypes carrying a Val at position 86 in the DR beta chain were increased in PP MS. The present study indicates that MS is genetically heterogeneous and shows a combined effect of HLA-DR and IL-1ra genes in susceptibility to the R/R form of the disease. PMID- 9413275 TI - Aging-associated changes in human brain. AB - A wide variety of anatomic and histological alterations are common in brains of aged individuals. However, identification of intrinsic aging changes--as distinct from changes resulting from cumulative environmental insult--is problematic. Some degree of neuronal and volume loss would appear to be inevitable, but recent studies have suggested that the magnitudes of such changes are much less than previously thought, and studies of dendritic complexity in cognitively intact individuals suggest continuing neuronal plasticity into the eighth decade. A number of vascular changes become more frequent with age, many attributable to systemic conditions such as hypertension and atherosclerosis. Age-associated vascular changes not clearly linked to such conditions include hyaline arteriosclerotic changes with formation of arterial tortuosities in small intracranial vessels and the radiographic changes in deep cerebral white matter known as "leukoaraiosis." Aging is accompanied by increases in glial cell activation, in oxidative damage to proteins and lipids, in irreversible protein glycation, and in damage to DNA, and such changes may underlie in part the age associated increasing incidence of "degenerative" conditions such as Alzheimer disease and Parkinson disease. A small number of histological changes appear to be universal in aged human brains. These include increasing numbers of corpora amylacea within astrocytic processes near blood-brain or cerebrospinal fluid brain interfaces, accumulation of the "aging" pigment lipofuscin in all brain regions, and appearance of Alzheimer-type neurofibrillary tangles (but not necessarily amyloid plaques) in mesial temporal structures. PMID- 9413276 TI - White matter neuronal heterotopia in temporal lobe epilepsy: a morphometric and immunohistochemical study. AB - A frequent abnormality in temporal lobes (TL) resected for pharmacoresistant epilepsy is the presence of heterotopic neurons within white matter (WM). We compared heterotopic neuron density in 22 TLs surgically resected from epilepsy patients with TLs from 22 non-neurologic cases obtained at autopsies. Neuronal density was assessed on LFB-PAS-stained and parallel sections immunoreacted for microtubule-associated-protein-2 (MAP-2). The white matter area was outlined by an image analysis system. Neurons, identified by morphologic features, were counted within the marked area. Results are expressed as mean +/- SD neurons/mm2. LFB/PAS sections: Epilepsy cases 4.11 +/- 1.86 Autopsy (normal) 2.35 +/- 0.96; MAP-2 sections: Epilepsy cases 4.08 +/- 1.22, autopsy (normal) 1.68 +/-0.92 (significant at 0.05 level by Wilcoxon's Rank Sums test). The lower number of MAP 2-immunopositive neurons in the control group as compared with the histologically identified group is most likely the result of antigen degradation resulting from an increased postmortem interval. These results indicate that normal TLWM contains a heterotopic population of neurons, and that this neuronal density is significantly higher in epilepsy patients. It is felt that this increased neuronal density is an epiphenomenon rather than the cause of seizures, and may be the result of decreased white matter either secondary to disruption of myelination, or loss of neurons as part of mesial temporal sclerosis. PMID- 9413277 TI - Expression of different isoforms of nitric oxide synthase in experimentally denervated and reinnervated skeletal muscle. AB - Denervated muscle fibers express enhanced levels of stress and apoptosis associated proteins and undergo apoptosis. In experimentally denervated and reinnervated rat facial muscle, we now evaluate changes in the expression patterns of different isoforms of nitric oxide synthase (NOS)-generating nitric oxide (NO), which mediates oxidative stress and apoptosis. Physiological expression of NOS corresponds to a constant sarcolemmal staining pattern for neuronal NOS (nNOS) and a patchy sarcolemmal and weak sarcoplasmic labeling for the endothelial NOS-isoform, with no expression for inducible NOS (iNOS). Denervated muscle displayed distinct downregulation of nNOS with preserved expression of dystrophin. Also, denervated and immediately reinnervated muscle fibers showed decreased expression of nNOS. However, muscle fibers reinnervated for 10 weeks revealed a restored physiological expression of nNOS. There were no changes in the expression of endothelial and inducible NOS. As NO is known to induce growth arrest and collapse of neuronal growth cones, downregulation of NOS may contribute to promotion of axonal regeneration by aiding formation of new endplates. NO is upregulated in reinnervated muscle fibers and thus prevents polyneural hyperinnervation by extrajunctional synapses. Furthermore, downregulation of NOS during denervation is compatible with the finding that low levels of NO contribute to apoptosis instead of necrosis in disease states of oxidative stress. PMID- 9413278 TI - BDNF attenuates functional and structural disorders in nerves of galactose-fed rats. AB - Galactose intoxication of rats was used to disrupt metabolism of Schwann cells and skeletal muscle, two sites that contain the polyol-forming enzyme aldose reductase (AR). Galactose-fed rats develop a neuropathy characterized by nerve conduction deficits and axonal atrophy. To investigate the possibility that galactose metabolism by AR influences axonal function and structure by altering production of neurotrophic factors, the impact of galactose intoxication on nerve and muscle BDNF levels and the effects of exogenous BDNF treatment on galactose neuropathy were examined using biochemical, electrophysiologic and morphometric techniques. Galactose feeding increased BDNF protein in peripheral nerve and muscle. Exogenous BDNF treatment attenuated motor nerve conduction velocity deficits in the sciatic nerve of galactose-fed animals and myelin splitting of motor axons in the ventral root. In contrast, sensory nerve conduction velocity (SNCV) deficits in the sciatic nerve and myelin splitting in the central projections of sensory neurons were not prevented by BDNF treatment. BDNF treatment did not attenuate reduced axonal caliber in the sciatic nerve, but did ameliorate the diminution of the caliber of central sensory projections in the dorsal root. These findings point to the potential use of BDNF in the treatment of peripheral neuropathies. PMID- 9413279 TI - Hydrocephalus in mice infected with a Theiler's murine encephalomyelitis virus variant. AB - The etiology of hydrocephalus is never established in the majority of clinical cases, while various agents, nutritional deficiencies, and genetic factors have been shown to play a role. Viral infection has been recognized as one of the causative factors in the development of hydrocephalus. The wild-type DA strain of Theiler's murine encephalomyelitis virus (TMEV), which belongs to the family Picornaviridae, causes a chronic demyelinating disease in mice with viral persistence that resembles multiple sclerosis. We found that a DA virus variant, hydrocephalus 101 virus (H101 virus), caused hydrocephalus in mice, a condition previously never described for TMEV. To clarify the relationship between DA virus infection and hydrocephalus, we compared H101 virus and wild-type DA virus infection in mice. Using immunohistochemistry and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling (TUNEL), we found that during the acute phase of infection, H101 virus caused macrocephaly and meningitis with the presence of apoptosis, while parenchymal involvement was not evident. In contrast, wild-type DA virus caused an acute polioencephalomyelitis with parenchymal infection and apoptosis. During the chronic phase, H101 virus infection caused communicating hydrocephalus without viral persistence. No demyelination and little or no anti-TMEV antibodies were observed in H101 virus infected mice. Sequence analysis revealed that H101 virus had mutations in the 5'UTR and capsid protein coding region. Characterization of this new hydrocephalus model gives insight into the possible viral involvement in human hydrocephalus cases of obscure etiology. PMID- 9413280 TI - c-Jun, JNK/SAPK kinases and transcription factor NF-kappa B are selectively activated in astrocytes, but not motor neurons, in amyotrophic lateral sclerosis. AB - There is increasing evidence that oxidative damage plays a major role in amyotrophic lateral sclerosis (ALS), but how it contributes to motor neuron degeneration and astrocytic gliosis, two pathologic hallmarks of the disease, is unknown. A few studies have suggested that ALS motor neurons die via apoptosis and show upregulation of c-jun, an immediate early gene that is necessary for neuronal apoptosis. In order to elucidate the mechanisms of cell damage induced by oxidant stress, we have studied in ALS and control spinal cord the immunohistochemical expression of c-Jun, of JNK/SAPK, a kinase that activates c Jun following various types of stress, and of NF-kappa B, a transcription factor that is induced by oxidant stress and has prominent neuroprotective functions. An in situ end-labeling assay was performed for detecting apoptotic cells. We show that (a) the JNK/SAPK-c-Jun pathway is dramatically overexpressed in ALS spinal cord; (b) the strongest activation occurs in astrocytes, while motor neurons show unusually low expression of the pathway; (c) increased JNK/SAPK expression in glial cells is accompanied by NF-kappa B activation, indicating the presence of a protective response to oxidant sress, which is deficient in motor neurons; (d) activation of JNK/SAPK, c-Jun and NF-kappa B is unrelated to apoptotic cell death. These results support the view that astrocytes are directly involved in the pathologic process of ALS, and might explain the selective vulnerability of motor neurons by their relative lack of antioxidant defenses. PMID- 9413281 TI - Spinal cord neuropathology in rat experimental autoimmune encephalomyelitis: modulation by oral administration of myelin basic protein. AB - Experimental autoimmune encephalomyelitis (EAE) is an inflammatory disease of the central nervous system (CNS) in which clinical neurological signs and histopathologic changes of disease can be suppressed by feeding CNS myelin proteins. Using immunohistochemistry and image analysis, the cellular immune response was quantified over the rostral-caudal axis of the spinal cord in rats with EAE and in animals fed high- or low-dose myelin basic protein (MBP) prior to inducing EAE (tolerized animals). In a subset of rats, MBP was fed 9 days after MBP immunization to examine the effect of oral tolerance on the progression of CNS pathology. In unfed rats or rats fed vehicle only, activated microglia and macrophages were co-localized with T-lymphocytes throughout the spinal cord, but greater cellular reactions were evident in gray matter relative to white matter. In all tolerized animals, the CNS inflammatory response was reduced relative to controls. Subtle pathologic changes were occasionally observed in the CNS of MBP fed animals, but the distribution of inflammatory cells in the dorso-ventral axis was more polarized in animals fed high-dose MBP. In this group, more T-cells and activated microglia were present in the dorsal spinal cord, specifically in the gray matter. In the group fed MBP after disease induction, clinical disease progressed as in control non-fed rats, but recovery from disease appeared to be accelerated. Thus, the results presented here provide a comprehensive analysis of the distribution and magnitude of inflammatory cells within the spinal cord in EAE and challenge the theory that MBP-induced EAE is only a white matter disease. These data also describe how the activation and distribution of immune effector cells is altered by oral tolerance and may help predict a range of neurological deficits not previously appreciated in EAE, particularly those effected by gray matter pathology. PMID- 9413282 TI - An immunohistochemical study of radial glial cells in the mouse brain prenatally exposed to gamma-irradiation. AB - The features of a glial cell population in the developing brain of mice prenatally exposed to 60Co gamma-irradiation at the most radiosensitive stage were studied with immunohistochemistry for anti-midkine (MK), anti-vimentin (Vim), and anti-GFAP antibodies. Anti-MK- and anti-Vim-positive radial glial fibers distributed in a similar radial fashion; these fibers were observed primarily in the embryonic period and disappeared after birth. Anti-MK- and anti Vim-stained radial fibers ran perpendicular to the pial surface in controls, whereas such fibers were disorganized 6 hours (h) after irradiation. This finding provided new evidence that the migratory pathways of young neurons were interrupted beginning a few hours after irradiation. By E17 the ectopic cell masses formed so as to replace the parts of the ventricular zone where no anti-MK immunoreactive radial fibers were present, but where anti-GFAP-stained fibrillary astrocytes emerged in the ectopic cell masses from the early postnatal period. The results suggested a twofold source of the generated astrocytes: either directly from a separate precursor of the astrocytes, or due to the transformation of the classic radial glial cells. In the newborn, numerous protoplasmic transitional forms displaced by astrocytes in irradiated brains indicated that reactive gliosis was a powerful response of a brain exposed to irradiation. PMID- 9413283 TI - Human telomerase RNA expression and MIB-1 (Ki-67) proliferation index distinguish hemangioblastomas from metastatic renal cell carcinomas. AB - Hemangioblastomas are low-grade, capillary rich neoplasms of the cerebellum and spinal cord that can occur sporadically or in the setting of Von Hippel-Lindau syndrome. The present study analyzed the utility of proliferation potential in differentiating hemangioblastoma from RCC metastatic to the central nervous system using a MIB-1 (Ki-67) labeling index and assessment of expression of the RNA component of telomerase. Immunohistochemical analysis for epithelial membrane antigen (EMA) and MIB-1 was performed on paraffin-embedded sections of 27 hemangioblastomas and 5 RCC metastatic to the central nervous system. All but one hemangioblastoma demonstrated low or negative MIB-1 immunoreactivity, while 4 of 5 RCC metastases had moderate or high labeling indices. Telomerase RNA expression was assessed in 10 hemangioblastomas and in all 5 metastatic RCC by in Situ hybridization. All 10 hemangioblastomas demonstrated a lack of expression of telomerase RNA, while all 5 metastatic RCCs showed moderate to strong expression. Our results suggest that the MIB-1 labeling index is useful in differentiating hemangioblastoma from metastatic RCC and assessment of telomerase expression can also provide novel information on the difference in growth potential of these tumors. PMID- 9413284 TI - The amyloid beta protein induces oxidative damage of mitochondrial DNA. AB - Multiple lines of evidence suggest involvement of oxidative stress in the pathogenesis of Alzheimer disease (AD). The finding that amyloid beta peptide (A beta) has neurotoxic properties and that such effects are mediated in part by free-radicals has provided an avenue to explore new therapeutic strategies. In this study, we showed that exposure of PC 12 cells to an A beta fragment induces oxidative damage of mitochondrial DNA. Cells were exposed for 24 h to 50 microM A beta (25-35) or to 50 microM of a control peptide with a scrambled sequence. Oxidative damage of mitochondrial DNA (mtDNA) was assessed using a Southern blot technique and an mtDNA-specific probe recognizing a 13.5-kilobase restriction fragment. Treatment of DNA with NaOH was used to reveal abasic sites and single strand breaks. Treatment with endonuclease III or FAPy glycosylase was used to detect pyrimidine or purine lesions, respectively. Cells exposed to A beta exhibited marked oxidative damage of mtDNA as evidenced by characteristic changes on Southern blots. Cells exposed to the scrambled peptide did not show such modifications. Simultaneous addition of the pineal hormone melatonin consistently prevented the A beta-induced oxidative damage to mtDNA. Mitochondrial dysfunction in AD has been demonstrated by several laboratories. This study provides experimental evidence supporting a causative role of A beta in mitochondrial lesions of AD. PMID- 9413286 TI - Neuropathology: art and science. AB - So many examples of computer-designed (rather than human-designed) displays occurred at a recent meeting of the American Association of Neuropathologists that we were stimulated to develop simple guides to help improve presentation. Various color combinations provide examples of the best (and worst) contrast between the message and the medium (background). PMID- 9413285 TI - Diffuse plaques in the striatum in Alzheimer disease (AD): relationship to the striatal mosaic and selected neuropeptide markers. AB - Although neuritic and diffuse plaques generally coexist in Alzheimer disease (AD) neocortex, the predominance of diffuse plaques in the striatum prompted us to explore the potential influence of striatal features such as the striatal mosaic on beta-amyloid (A beta) deposition. Using double immunohistochemistry with an antibody to A beta in combination with antibodies to met-enkephalin or somatostatin, we investigated the relationship between diffuse plaques and neuropeptide distribution in the dorsal striatum. This relationship was examined in "pure" AD cases as well as in AD cases with coexisting Parkinson disease (PD) pathology, i.e. nigral degeneration and Lewy bodies at any site (AD + PD). Despite the presence of numerous A beta-positive diffuse plaques in both groups, the mosaic pattern, as exemplified by met-enkephalin-immunoreactive patches, was preserved. No obvious association was observed between the plaques and met enkephalin-positive patches or somatostatin-immunoreactive neurons. Tyrosine hydroxylase immunoreactivity in the matrix was, however, diminished in AD + PD, most likely reflecting the nigral degeneration in these cases. Overall, these observations suggest that neostriatal A beta deposition in AD is not influenced by environmental factors associated with the striatal mosaic. PMID- 9413287 TI - Dr. William Worrall Mayo's gynecologic surgical practice. PMID- 9413288 TI - Variability in anticoagulation control predicts thromboembolism after mechanical cardiac valve replacement: a 23-year population-based study. AB - OBJECTIVE: To assess optimal control of blood anticoagulation to maximize antithrombotic protection after mechanical cardiac valve replacement. DESIGN: A population-based study of 96 patients with a mean follow-up of 7.7 years (range, 1 month to 23 years) was performed in Olmsted County, Minnesota, and 10,301 prothrombin time (PT) ratios were determined after mechanical heart valve replacement. MATERIAL AND METHODS: PT ratios were analyzed in a new time dependent Cox proportional-hazards model by defining an algorithm for comparing variability in PT ratios at each month of follow-up and relating these to thromboembolic events. The new method was compared with several conventional time independent definitions. RESULTS: During 740 person-years of follow-up, 19 of 96 patients (20%) had 27 thromboembolic events. Of these 19 patients, 8 (42%) had events within 3 months after valve replacement. Freedom from any thromboembolic event was 72% at 15 years. The event rate was high (7.5% per year) during high variability and low (0.9% per year) during low variability in the PT ratio. This relationship was lost when time dependence was removed. More PT ratios were less than 1.5 during high (27%) than during low (19%) variability. Several conventional definitions of adequacy of anticoagulation that averaged PT ratios before a thromboembolic event or throughout follow-up or that compared the proportion of PT ratios above or below a fixed ratio did not define or only partially defined different thromboembolic risks. CONCLUSION: Periods of high and low variability of PT ratios define high and low risk of thromboembolism, respectively. PMID- 9413289 TI - Increased parathyroid hormone-related peptide in patients with hypercalcemia associated with islet cell carcinoma. AB - OBJECTIVE: To report the high prevalence of increased parathyroid hormone-related peptide (PTHrP) in patients with islet cell carcinoma and associated hypercalcemia. DESIGN: We conducted a retrospective study of PTHrP levels in patients with hypercalcemia and eucalcemia associated with islet cell carcinoma and compared these findings with those in healthy subjects. MATERIAL AND METHODS: Using a sensitive PTHrP immunochemiluminometric assay, we measured PTHrP levels in 17 patients with islet cell carcinoma and 110 healthy subjects. The differences between PTHrP levels in patients with normal and those with high serum calcium concentrations were analyzed statistically. RESULTS: PTHrP levels were significantly higher (P < 0.01) in 10 patients with hypercalcemia and islet cell carcinoma (median, 14.0 pmol/L; range, undetectable to 40.1) than in 7 patients with eucalcemia and islet cell carcinoma (median, undetectable; range, undetectable to 1.3 pmol/L) or in the 110 healthy subjects (median, undetectable; range, undetectable to 4.2 pmol/L). The range of increased PTHrP levels in hypercalcemic islet cell carcinoma was 2 to 20 times the upper normal limit (2.0 pmol/L). Decreased PTHrP and serum calcium and increased parathyroid hormone levels were demonstrated in two patients after effective therapy. For all seven eucalcemic patients with islet cell carcinoma, PTHrP levels did not differ significantly from those in healthy subjects. CONCLUSION: PTHrP levels are increased in a substantial proportion of patients with hypercalcemia and islet cell carcinoma and seem to decrease after treatment of the underlying tumor. Measurement of PTHrP levels may be useful for confirming the diagnosis of hypercalcemia associated with malignant disease and for monitoring of therapy. PMID- 9413290 TI - Telemedicine and the diagnosis of speech and language disorders. AB - OBJECTIVE: To summarize results of telemedicine evaluations of speech and language disorders in patients in a small, rural hospital and in large multidisciplinary medical practices. MATERIALS AND METHODS: Eight patients underwent assessment as part of experiments with the National Aeronautics and Space Administration-launched Advanced Communications Technology Satellite. A second clinician was on-site with patients to assess the reliability of satellite observations. Twenty-four previously videotaped samples of speech disorders were also transmitted to assess agreement with original face-to-face clinical diagnoses. In addition, results of 150 telemedicine evaluations among Mayo Clinic practices in Minnesota, Arizona, and Florida were examined retrospectively. RESULTS: Evaluations were reliable, and patient satisfaction was good. Diagnoses were consistent with lesion localization and medical diagnosis when they were known, and they frequently had implications for lesion localization and medical diagnosis and management when they were previously unknown. The frequency of uncertain diagnosis (13%) for evaluation among the Mayo practices was only slightly higher than that encountered in face-to-face practice. Face-to-face evaluations were considered necessary for only 6 of the 150 patients (4%). CONCLUSION: Telemedicine evaluations can be reliable, beneficial, and acceptable to patients with a variety of acquired speech and language disorders, both in rural settings and within large multidisciplinary medical settings. PMID- 9413291 TI - Subacute diabetic proximal neuropathy. AB - OBJECTIVE: To evaluate the clinical, electrophysiologic, autonomic, and neuropathologic characteristics and the natural history of subacute diabetic proximal neuropathy and its response to immunotherapy. MATERIAL AND METHODS: For the 12-year period from 1983 to 1995, we conducted a retrospective review of medical records of Mayo Clinic patients with diabetes who had subacute onset and progression of proximal weakness. The responses of treated versus untreated patients were compared statistically. RESULTS: During the designated study period, 44 patients with subacute diabetic proximal neuropathy were encountered. Most patients were middle-aged or elderly, and no sex preponderance was noted. The proximal muscle weakness often was associated with reduced or absent lower extremity reflexes. Associated weight loss was a common finding. Frequently, patients had some evidence of demyelination on nerve conduction studies, but it invariably was accompanied by concomitant axonal degeneration. The cerebrospinal fluid protein concentration was usually increased. Diffuse and substantial autonomic failure was generally present. In most cases, a sural nerve biopsy specimen suggested demyelination, although evidence of an inflammatory infiltrate was less common. Of 12 patients who received treatment (with prednisone, intravenous immune globulin, or plasma exchange), 9 had improvement of their conditions, but 17 of 29 untreated patients (59%) with follow-up also eventually had improvement, albeit at a much slower rate. Improvement was usually incomplete. CONCLUSION: We suggest that the entity of subacute diabetic proximal neuropathy is an extensive and severe variant of bilateral lumbosacral radiculoplexopathy, with some features suggestive of an immune-mediated cause. It differs from chronic inflammatory demyelinating polyradiculoneuropathy in that most cases have a more restricted distribution and seem to be monophasic and self limiting. The efficacy of immunotherapy is unproved, but such intervention may be considered in the severe and progressive cases or ones associated with severe neuropathic pain. PMID- 9413292 TI - Acute hepatitis E by a new isolate acquired in the United States. AB - OBJECTIVE: To report the first case of acute hepatitis E by a novel isolate acquired in the United States and confirmed by nucleotide sequencing. MATERIAL AND METHODS: We describe the clinical manifestations and the results of associated laboratory studies in a man who was found to have acute hepatitis E infection. RESULTS: A 62-year-old man was hospitalized because of fever, abdominal pain, and jaundice. After an initial evaluation did not provide a cause, his serum was found to be positive for IgG anti-hepatitis E virus (HEV) by three antibody assays. Serum was also positive for HEV RNA by reverse transcriptase polymerase chain reaction (PCR). Sequencing results from the PCR products demonstrated substantial differences at the nucleotide level between this strain and the known Mexican and Burmese strains. CONCLUSION: On the basis of this initial report, HEV should be considered an etiologic agent in patients with acute non-ABC hepatitis in the United States. PMID- 9413293 TI - Ursodeoxycholic acid delays the onset of esophageal varices in primary biliary cirrhosis. AB - OBJECTIVE: To address the effect of ursodeoxycholic acid therapy on development of esophageal varices in patients with primary biliary cirrhosis. MATERIAL AND METHODS: We compared, as part of a prospective treatment trial, the risk of varices developing in patients with primary biliary cirrhosis who received ursodeoxycholic acid (13 to 15 mg/kg daily) versus those who received placebo for up to 4 years. Upper endoscopy was performed every 2 years or as indicated clinically. At the end of the 4-year period, all patients in the placebo group were offered ursodeoxycholic acid therapy. During follow-up, the risk of developing endoscopically confirmed varices was assessed. RESULTS: The 180 patients who entered the ursodeoxycholic acid trial were assessed for the presence or absence of varices by esophagogastroduodenoscopy; 139 patients had no varices, and 41 patients demonstrated varices on initial examination. At 4 years, the risk of newly developing endoscopically confirmed varices was 16% for the ursodeoxycholic acid-treated patients and 58% for the placebo-treated patients (P < 0.001). Thus, the use of ursodeoxycholic acid was associated with a significantly lower risk of developing varices in patients with primary biliary cirrhosis. CONCLUSION: In addition to biochemical improvement, delay in death, and prolongation of time to orthotopic liver transplantation, ursodeoxycholic acid has now been demonstrated to decrease the risk of esophageal varices developing in patients with primary biliary cirrhosis. PMID- 9413294 TI - Allergic contact dermatitis due to topical application of corticosteroids: review and clinical implications. AB - Allergy due to topically applied corticosteroids is being recognized more frequently. Testing for hypersensitivity to these agents is performed with delayed hypersensitivity patch testing. Cross-reactivity among topically administered corticosteroids is frequent and often can be predicted on the basis of additional patch testing and an established classification scheme. Herein we review allergy due to topically applied corticosteroids with regard to its prevalence, means of testing, cross-reactivity among the subclasses, risk factors, and relationship to steroids. PMID- 9413295 TI - Lupus and pulmonary nodules consistent with bronchiolitis obliterans organizing pneumonia induced by carbamazepine. AB - Carbamazepine-induced lupus is uncommon; its frequency is less than 0.001% of the cases of lupus treated. Herein we describe a 52-year-old woman who had development of facial erythema, arthralgia, dyspnea, and multiple pulmonary rounded masses and nodules while she was taking carbamazepine for epilepsy. Pulmonary histologic examination showed bronchiolitis obliterans organizing pneumonia. Antinuclear antibodies and antihistone antibodies were present without antibodies to double-stranded DNA. Thirteen months after carbamazepine had been withdrawn, all symptoms had disappeared without the use of anti-inflammatory drugs. Even though pulmonary involvement is possible during carbamazepine treatment, to our knowledge, bronchiolitis obliterans organizing pneumonia on its own or associated with carbamazepine-induced lupus has not been previously described. PMID- 9413296 TI - Methysergide-induced retroperitoneal fibrosis: successful outcome and two new laboratory features. AB - A 25-year-old woman sought medical attention because of iliocaval manifestations of retroperitoneal fibrosis while she was taking methysergide. Laboratory studies yielded substantially increased serum procollagen III levels and anticardiolipin antibodies accompanied with anti-beta(2) glycoprotein I, findings not previously described with this disorder. Clinical and laboratory manifestations resolved after cessation of methysergide therapy. PMID- 9413297 TI - Intravenous immunoglobulin-induced lichenoid dermatitis: a unique adverse reaction. AB - Intravenous immunoglobulin (IVIG) therapy has been used to treat various diseases. Generalized allergic reactions (types I, II, and III) to IVIG are uncommon and are usually related to the rate of infusion. Cutaneous reactions have been anecdotally described. Herein we describe a 73-year-old man with Sjogren's syndrome who had development of a lichenoid cutaneous eruption after administration of IVIG therapy. CD3 immunostaining demonstrated the overwhelming presence of T lymphocytes within a lesional skin biopsy specimen, and thus we present an alternative cell-mediated immune mechanism (type IV) by which allergic reactions to IVIG may occur. PMID- 9413298 TI - Bengt Samuelsson--Nobel Prize winner for studies of prostaglandins, thromboxanes, and leukotrienes. PMID- 9413299 TI - Carcinoma of the stomach with hepatocyte differentiation (hepatoid adenocarcinoma). AB - A case of hepatoid adenocarcinoma of the stomach is reported, and the literature is reviewed. The stomach is one of the most common sites in which hepatoid adenocarcinomas have been detected. Of the 59 cases reviewed from the literature (including the current case), a 2:1 male predominance was noted, and the serum alpha-fetoprotein level was almost always increased. All patients were adults, and most had evidence of metastases at the time of resection. Prognosis seems less favorable than that associated with the more common intestinal type of adenocarcinoma of the stomach. Hepatoid adenocarcinomas typically show periodic acid-Schiff-positive, diastase-resistant intracytoplasmic globules, which are demonstrated to be positive with antibodies to alpha-fetoprotein. The tumor cells resemble liver cells, and rare cases, including our own, have evidence of bile production. In our case, messenger RNA for albumin, unique to liver cells, was demonstrated by in situ hybridization of the tumor cells. PMID- 9413300 TI - Evaluation and management of amenorrhea. AB - All women who enter menopause experience amenorrhea unless they receive hormone replacement therapy. In younger women, amenorrhea unrelated to pregnancy and lactation can be a distressing symptom. In addition to its psychologic morbidity, amenorrhea may be the manifesting feature of a wide array of anatomic and endocrine abnormalities. Amenorrhea results in impaired fertility. When estrogen levels are low, changes in mineral, glucose, and fat metabolism accompany amenorrhea. These metabolic changes affect bone and cardiovascular health, increasing the risk of osteoporosis and coronary heart disease in later life. Amenorrhea with hyperandrogenism, most commonly caused by the polycystic ovarian syndrome, may cause endometrial hyperplasia and increases the risk of endometrial adenocarcinoma. Because of the broad differential diagnosis of amenorrhea, establishing an accurate diagnosis can prove challenging. In this article, we outline one approach to the assessment of patients with amenorrhea and to the management of its common causes and consequences. PMID- 9413301 TI - 80-Year-old woman with dementia and parkinsonism. PMID- 9413302 TI - Bladder dysfunction and management in multiple sclerosis. AB - Symptomatic bladder dysfunction occurs at some time in most patients with multiple sclerosis. The relapsing-remitting course and progressive loss of mobility associated with multiple sclerosis make management of urinary urgency and incontinence difficult. Urodynamic evaluation serves as a guideline for appropriate treatment. After accurate diagnosis of bladder dysfunction, a management program is developed with use of fluid schedules, voiding techniques, neuropharmacologic manipulation, intermittent catheterization, surgical treatment, and other adjunctive measures as indicated. The goals of treatment are to protect and preserve renal function, relieve symptomatic voiding dysfunction, and avoid subsequent urinary complications. A management program should be individualized, dynamic, and monitored with periodic, systematic urologic review to maintain these goals. PMID- 9413303 TI - Management of impairment, disability, and handicap due to multiple sclerosis. AB - In this article, we update management measures for patients with multiple sclerosis (MS) that can improve or prevent impairment, disability, and handicap and include those factors that a primary-care physician can implement or facilitate. The medical literature since 1989 was reviewed. Although new drug trials hold promise to decrease impairment from MS, well-coordinated interdisciplinary care to minimize disability and handicap most profoundly affect the quality of life for patients with MS. MS is usually not severely disabling, and appropriately timed intervention can prevent secondary impairment and reduce disability and handicap. Pharmacologic, physical, and psychosocial issues- ranging from spasticity, pain, weakness, and tremor to neurogenic bowel management and sexuality--are addressed. General wellness measures remain important. The influence of the Americans With Disabilities Act is discussed, and specific adaptive equipment and social resources are outlined. The ultimate goals of management of patients with MS are functional independence and efficient use of medical and community resources: a focus on "ability" rather than "disability." Although impairment can limit function, wellness and adjustment have no boundaries. PMID- 9413305 TI - The literature on quality of life and organizational randomized clinical trials. PMID- 9413304 TI - Hepatitis E in the United States: a case of "hog fever"? PMID- 9413306 TI - Medicare treatment differences for blacks and whites. AB - OBJECTIVES: This study investigated racial differences in procedure use among elderly Medicare beneficiaries. It is hypothesized that providers do not discriminate inappropriately in treating black and white patients and that the apparent differences in black-white treatment could be attributed to other differences between the two populations. METHODS: Rates of use for selected procedures were examined among two patient groups: (1) the universe of Medicare beneficiaries in 10 states and the District of Columbia and (2) a subset of this sample created by matching beneficiaries on the basis of zip code of residence to neutralize the effects of black-white differences in provider access and regional practice patterns. Because all Medicare beneficiaries have a common core of standard benefits, the importance of financial access differences in accounting for black/white utilization differences is diminished. RESULTS: Three major findings were indicated from this study: (1) area-controlled comparisons find even larger black-white disparities than those shown from uncontrolled comparisons, (2) the disparities are larger in southern states, and (3) the disparities vary substantially with procedure cost. CONCLUSIONS: Although no clinical data were analyzed, providers appeared to be giving less intensive treatment to otherwise similar black Medicare beneficiaries. PMID- 9413307 TI - The impact of health maintenance organization penetration on the use of hospitals that serve minority communities. AB - OBJECTIVES: Health maintenance organization (HMO) penetration has made hospital markets more price competitive. Hospitals in minority communities may be at a competitive disadvantage because they serve patients who are, on average, sicker and more likely to be uninsured or underinsured. This study estimated the impact of HMO penetration on the use of hospitals in minority communities during 1987 to 1992. METHODS: Using a sample of 1,413 short-term general hospitals from the 85 largest metropolitan statistical areas, the determinants of hospitals' patient volumes were estimated. Hospitals located in predominately nonwhite neighborhoods were designated minority hospitals, and other hospitals were designated nonminority hospitals. Using regression analysis, the impact of HMO penetration and concentration on hospitals' patient volumes were estimated. By interacting the HMO penetration and concentration variables with a minority hospital indicator variable, HMOs' impact on minority hospitals was calculated. RESULTS: Health maintenance organization penetration was correlated with lower patient volumes in minority hospitals and higher patient volumes in nonminority hospitals. Competition in HMO markets was correlated with lower patient volumes for all hospitals. This effect was stronger for minority hospitals. CONCLUSIONS: These findings suggest that minority hospitals may be at risk of losing patients as HMO penetration increases. PMID- 9413308 TI - Breast screening behavior and interactions with health care providers among lower income women. PMID- 9413309 TI - The effects of ethnicity and language on medical outcomes of patients with hypertension or diabetes. PMID- 9413310 TI - Time trends in late-stage diagnosis of cervical cancer. Differences by race/ethnicity and income. PMID- 9413311 TI - A strategy in plain sight. PhyCor says clues to its surprise purchase of MedPartners were there all along. PMID- 9413312 TI - The Medicare mall. Will Washington like what it built? AB - "Shopping spree" doesn't begin to describe it. Next year, 38 million seniors will begin choosing their health care coverage from a menu crammed with options. Medicare Plus Choice means just that: HMOs, provider-sponsored plans, private-pay options, medical savings accounts, and more. It also means a bevy of new rules- and risks--for everyone. PMID- 9413313 TI - Operation start-up. AB - When Mount Carmel Health System sized up the Medicare market in Columbus, Ohio, executives liked what they saw: Seniors trusted them more than insurers to run a managed care plan. Only six months later, their provider-sponsored plan opened for business under a HCFA pilot project. Here's how Mount Carmel pulled it off. PMID- 9413314 TI - Happy is as happy does? AB - Good food and clean rooms go only so far. Patient satisfaction surveys are asking tough questions about quality of care. What's more, standard national surveys may be coming sooner than you think. PMID- 9413315 TI - Hospital conversions. Good deal, but there's a catch. PMID- 9413316 TI - Mergers. Big ... bigger ... biggest. PMID- 9413317 TI - Advocacy. Charity champion. PMID- 9413318 TI - Bond ratings. Where credit is due. PMID- 9413319 TI - The economics of nursing home practice. PMID- 9413320 TI - Avoiding heparin dosing mistakes. PMID- 9413321 TI - Antinuclear antibody testing. PMID- 9413322 TI - Bulletin on women's health. PMID- 9413323 TI - Ulcerative colitis. PMID- 9413324 TI - Uses of a Kock pouch. PMID- 9413325 TI - Considering Humalog. PMID- 9413326 TI - Breakthrough pain with the fentanyl patch. PMID- 9413327 TI - When a patient goes 'AMA'. PMID- 9413328 TI - Nurse-patient boundaries: crossing the line. PMID- 9413329 TI - Emergency! Meningococcal disease. PMID- 9413330 TI - Basic assessment series: the adult cardiovascular system. PMID- 9413331 TI - How patients die. PMID- 9413332 TI - Clinical snapshot: mucositis. PMID- 9413333 TI - Technology scorecard: focus on infection control. PMID- 9413334 TI - Tillie is going home. PMID- 9413335 TI - Infection control challenges during hospital renovation. PMID- 9413336 TI - Giving an outstanding presentation. PMID- 9413337 TI - Message received. PMID- 9413338 TI - Our safety is important, too. PMID- 9413339 TI - Are academic medical societies needed in a changing healthcare arena? AB - Medical research has been demonstrated to increase the quality of life for all Americans and to be a sound investment with clear financial savings. However, academic health centers, which are the major source of medical research, have experienced continued erosion of their financial base of support. The author reviews some of the elements which have precipitated the current crisis for many academic health centers and proposes short- and long-term goals that should be endorsed by academic medical societies as a means of reducing these institutions' financial dependence on clinical income and restoring to them their primary social mission: to train the highest caliber of practicing physicians and to be centers of fundamental, not technological, research. Such research has been demonstrated to reduce, not increase, health care costs. Academic medical societies must work together to educate the public and Congress about the needs of academic health care centers, and to provide a cohesive and constructive set of practical recommendations that can strengthen their future financial stability. PMID- 9413340 TI - Nasal mucosal cell alterations in HIV-infected patients. AB - Patients with HIV infection often have nasopharyngeal symptoms related to inflammatory or infectious diseases of the upper respiratory tract. In this study, we examined specific nasal mucosal cytologic alterations in adult patients with HIV infection for associations with nasopharyngeal symptoms and other clinical parameters. Mucosal cytology was obtained in 62 patients from an urban HIV clinic using a plastic curettelike probe. The quantities of goblet cells, vacuolated cells, and leukocytes were determined and analyzed for associations with various clinical aspects of these patients and specifically with the presence or absence of prolonged (> 2-week duration) nasopharyngeal symptoms. Goblet cell, but not vacuolated cell, increases were observed in samples in which nasal eosinophilia was present and in samples obtained from April to October without specific associations with nasopharyngeal symptoms or histories of atopic disease. A history of allergic rhinitis or recent upper respiratory infection was significantly associated with increased proportions and total numbers of epithelial cells that showed vacuolization. In patients with prolonged nasopharyngeal symptoms, significantly higher numbers of nasal leukocytes were observed but higher proportions of vacuolated cells were not. Scores of tests for abnormal physical findings in the nose were higher for patients with prolonged nasopharyngeal symptoms than for those without. Peripheral blood CD4 concentrations, gender, nasal substance abuse history, and other comorbidities did not influence either vacuolated cell or goblet cell quantities. These data show that prolonged nasopharyngeal symptoms in HIV infection are associated with a certain nasal cellular pattern. It is conceivable that this pattern relates to recurrent or prolonged nasal inflammation secondary to upper respiratory infection. PMID- 9413341 TI - Renal responses to hypertonic saline infusion in salt-sensitive spontaneously hypertensive rats. AB - In Wistar Kyoto (WKY) and Sprague Dawley rats, high dietary sodium chloride (NaCl) increases natriuretic and diuretic responses to acute isotonic saline infusion, but in NaCl-sensitive spontaneously hypertensive rats (SHR-S), a high NaCl diet causes negligible increases in natriuretic and diuretic responses. To investigate whether this deficit in sodium and fluid excretion in SHR-S is stimulus (volume)-specific or because of a more generalized alteration in renal function, the present study measured, in SHR-S and Wistar Kyoto rats, natriuretic and diuretic responses to a hypertonic saline infusion (the amount of sodium infused was equal to that infused in a previous, isotonic experiment). Eight-week old Wistar Kyoto rats, SHR-S, and salt-resistant SHR were given a basal (1%) or high (8%)-NaCl diet for 2 weeks. Intravenous infusion of hypertonic saline increased mean arterial pressure and reduced heart rate in all groups. Baseline sodium excretion was lower in SHR-S compared with salt-resistant SHR with either diet, but after infusion of hypertonic saline, all 6 groups displayed significant increases in sodium and fluid excretion, glomerular filtration rate, and effective renal blood flow (ERBF). The percent-sodium excretion in response to hypertonic saline infusion was slightly, but significantly, lower in SHR-S (compared with salt-resistant SHR) for either the basal or the high-NaCl diet. We conclude that renal responses to hypertonic saline infusion are affected minimally in SHR-S compared with salt-resistant SHR or Wistar Kyoto rats. Therefore, the deficits in renal function observed in SHR-S after volume loading are not reflected in a renal deficit to hypertonic saline challenge. PMID- 9413342 TI - Effects of antihypertensive drugs on antioxidant enzyme activities and renal function in stroke-prone spontaneously hypertensive rats. AB - The reactive oxygen species has been proposed as a key mediator of the progression of renal injury associated with essential hypertension. Among the defense systems operating against the reactive oxygen species, superoxide dismutase, glutathione peroxidase, and catalase are the most important antioxidant enzymes (AOEs). In the present study, systolic blood pressure, renal function (creatinine clearance, urinary albumin, and N-acetyl-beta D glucosaminidase excretion), renal intrinsic AOE activities, and renal histopathology were determined in stroke-prone spontaneously hypertensive rats and Wistar Kyoto rats. The effects of a 20-week treatment using three antihypertensive drug regimens--captopril, a sulfhydryl-containing angiotensin converting enzyme inhibitor; temocapril, a potent, non-sulfhydryl-containing angiotensin-converting enzyme inhibitor prodrug; and a conventional triple drug combination that includes a vasodilator (hydralazine, hydrochlorothiazide and reserpine)--on renal function, renal tissue, AOE activities, and renal histopathologic abnormalities were evaluated in stroke-prone spontaneously hypertensive rats. Renal function and AOE activities were lower in the stroke prone spontaneously hypertensive rats than in the Wistar Kyoto rats. Normalization of systolic blood pressure using the antihypertensive drugs improved renal function and produced a nonuniform alteration in renal AOEs; only glutathione peroxidase activity increased significantly with the use of all three drug regimens. The mild renal histopathologic abnormality in stroke-prone spontaneously hypertensive rats was not altered by drug treatment. The improvement in renal function may be related to an increase in glutathione peroxidase activity, but no correlation was seen between renal function changes and alteration in activities of superoxide dismutase or catalase. PMID- 9413343 TI - Cardiovascular risk factors and behavior lifestyles of young women: implications from findings of the Bogalusa Heart Study. AB - The primary purposes of this article are to highlight important issues related to cardiovascular risk factors and behavior life-styles in young women and to examine racial (black-white) differences in risk factors that relate to cardiovascular disease. In childhood, some girls show cardiovascular risk factors of higher blood pressure levels, dyslipidemia, and obesity, all of which continue into young adulthood. Factors that contribute to abnormal risk factors are a high saturated fat diet, excess energy intake related to inactivity, and cigarette smoking. Trends of obesity are documented; and young white girls are continuing to use tobacco, more so than boys and black girls. Although the onset of clinical cardiovascular disease is delayed in women, the stage is set in childhood for the development of early cardiovascular risk. PMID- 9413344 TI - Pheochromocytoma crisis caused by contemporary ergotamine, caffeine, and nimesulide administration. AB - Pheochromocytoma is a rare tumor that secretes excess catecholamines. Pheochromocytoma crises may be precipitated by the use of several drugs. This article describes the case of a patient affected by pheochromocytoma in whom multiple organ failure developed after contemporary administration of ergotamine, caffeine, and nimesulide. The patient recovered completely long after surgical intervention. PMID- 9413345 TI - Peripartum cardiomyopathy occurring in a patient previously treated with doxorubicin. AB - We present a 28-year-old primigravida woman in whom congestive heart failure developed 3 months after delivery of a neonate. The patient underwent doxorubicin treatment 10 years previously. The combined cardiotoxicity of prior doxorubicin treatment and pregnancy is considered, and the importance of careful follow-up of cardiac function during pregnancy and postpartum in patients previously treated with doxorubicin is stressed. PMID- 9413346 TI - Recovery of pancreatic beta-cell function in hemochromatosis: combined treatment with recombinant human erythropoietin and phlebotomy. AB - A patient with diabetes mellitus caused by secondary hemochromatosis was treated using recombinant human erythropoietin and phlebotomy. A total of 12 g of iron had been infused in the patient because of iron deficiency anemia. Blood glucose level was 17.3 mmol/L, and hemoglobin A1c level was 9.0% at admission. He was treated using phlebotomy (400 mL per week), along with subcutaneous injection of 3,000 U of recombinant human erythropoietin three times a week. After approximately 100 days, a total of 5,500 mL of blood (2.75 g iron) could be removed. Serum ferritin level decreased from 10,000 micrograms/L to 4,807 micrograms/L. Fasting and maximum serum C-peptide immunoreactivity values during 100-g oral glucose tolerance tests were improved from 0.14 nmol/L to 0.42 nmol/L and from 1.84 nmol/L to 2.61 nmol/L, respectively. This case suggests that pancreatic beta-cell recovers in diabetes caused by hemochromatosis by reducing iron overload during a short period. PMID- 9413347 TI - Successful treatment with cyclosporine in a case of hemophagocytic syndrome manifesting as severe liver dysfunction. AB - Forms of hemophagocytic syndrome, which affects mainly children, vary from mild to very severe and often fatal. We describe an adult patient with hemophagocytic syndrome in whom severe liver dysfunction developed. The condition continued to deteriorate despite treatment with plasma exchange, high-dose gamma globulin, and corticosteroid therapy. Treatment with cyclosporine (2.3 mg/kg/day) dramatically improved the condition and normalized liver function. Cyclosporine reduced the serum levels of ferritin, interferon-tau, interleukin-6, and soluble interleukin 2 receptor. These findings suggest that hemophagocytic syndrome accompanied with severe liver dysfunction results from hypercytokinemia, and cyclosporine is useful in preventing a fatal outcome during the acute phase. PMID- 9413348 TI - Reversible hemiplegia as a consequence of severe hyperkalemia and cocaine abuse in a hemodialysis patient. AB - Severe hyperkalemia may cause weakness that typically is ascending and symmetric. In an isolated case report, hemiplegia occurred after the development of hyperkalemia in a patient with a known central nervous system lesion. We describe a patient requiring long-term hemodialysis who had near-fatal hyperkalemia, hemiplegia, and rhabdomyolysis after abuse of crack cocaine. The hemiplegia resolved after normalization of serum potassium using emergency dialysis. No brain lesion could be identified during computed tomography or by electroencephalography, and the patient had no residual neurologic deficits. We conclude that this patient had hemiplegia secondary to cocaine-induced cerebral vasoconstriction because no structural lesion could be found and that the neurologic deficit was worsened by severe hyperkalemia, which probably resulted from cocaine-induced rhabdomyolysis. Hence, despite the absence of a structural lesion of the brain, severe hyperkalemia, typically associated with symmetric, ascending muscle weakness, may contribute to causing focal weakness in the condition of cocaine-induced vasoconstriction. PMID- 9413349 TI - Hemodynamic changes of aortic regurgitation. AB - Hemodynamic changes associated with acute AR include a rapid increase in LV pressures during diastole, markedly elevated pressures at end diastole, and premature closure of the mitral valve. Systemic diastolic pressures may be low, but there is a minimal increase in pulse pressure. In very severe cases of acute AR, cardiac output may decrease, leading to hypotension. In chronic AR, the LV remodels to accommodate the regurgitant volume flow, and stroke volume increases to maintain effective forward blood flow. These adaptations lead to a dilated LV, a widened pulse pressure, and a low diastolic blood pressure, which are the classic findings of chronic AR. PMID- 9413350 TI - Southwestern Internal Medicine Conference. New advances in immunosuppression therapy for renal transplantation. PMID- 9413351 TI - Management of oropharyngeal trauma in children. AB - OBJECTIVE: To determine the indications for admission, requisite imaging studies, and urgent medical or surgical intervention. DESIGN: We retrospectively reviewed the charts of 26 children (age range, 5 months to 14 years) who were seen by the otolaryngology service in the emergency department at the Children's National Medical Center, Washington, DC, from 1985 to 1993 and who were diagnosed as having oropharyngeal trauma. We specifically looked for common findings in the history and physical examination on initial presentation to predict the necessary steps in evaluation and management. SETTING: Tertiary care pediatric referral center. RESULTS: Indications for admission were (1) concern about neurologic injury, (2) concern about vascular injury, (3) radiographic evidence of retropharyngeal free air or abscess, (4) pneumomediastinum, and (5) unreliable adult supervision at home. Six patients required surgery; 3 underwent retropharyngeal aspiration or incision and drainage procedures; 2 required neck explorations; and 1, who had an impaled foreign body in the parapharyngeal space, underwent surgical extraction. There were no vascular, neurologic, or other permanent injuries. CONCLUSIONS: Oropharyngeal trauma may result in palatal and posterior pharyngeal wall injury requiring closure of lacerations and management of retropharyngeal free air. Rarely does an injury lead to retropharyngeal abscess or significant pneumomediastinum. Lateral oropharyngeal injuries require increased concern about potential neurovascular impairment. However, neither the mechanism of injury nor the degree of injury correlates with the potential for neurovascular sequelae. Since neurovascular involvement may not become clinically apparent until days or weeks after the incident, admission for observation alone should be based on the distance from the patient's home to the hospital and on the level of reliable adult supervision. Indications for medical and surgical treatment of internal carotid artery thrombosis remain controversial. PMID- 9413352 TI - Relative incidence and alternative approaches for surgical drainage of different types of deep neck abscesses in children. AB - OBJECTIVES: To determine the relative frequency of retropharyngeal abscesses (RPAs) vs lateral pharyngeal abscesses (LPAs) and to analyze alternative approaches for surgical drainage. DESIGN: Retrospective chart review. SETTING: Tertiary care children's hospital. PATIENTS: Seventy pediatric patients who were evaluated, admitted, and treated for presumed deep neck abscesses (RPAs and LPAs) between January 1, 1986, and December 31, 1996. INTERVENTION: Intravenous antibiotic therapy and surgical drainage. MAIN OUTCOME MEASURE: Clinical resolution of the abscess. RESULTS: Fifty-eight patients were evaluated with computed tomographic scan. Thirteen of these patients did not have surgical intervention. Of 12 patients diagnosed as having an isolated RPA, all had intraoral surgical drainage and 9 had evidence of pus at surgery. Twenty-one patients had an isolated LPA. Sixteen of these underwent intraoral drainage and 5 underwent external drainage. Purulence was found at surgery in 14 and 2 patients, respectively. The remaining 12 patients had a combination of RPA and LPA. Eight patients underwent intraoral drainage, and 4 patients required both intraoral and external approaches. Purulence was found at surgery in 5 and 4 patients, respectively. Of the 12 patients who were not evaluated with computed tomographic scan, two thirds were treated prior to 1987. Six of these 12 patients underwent surgical drainage via an intraoral approach, and 4 of the 6 patients had pus. The remaining 6 improved without surgery. CONCLUSIONS: Most deep neck abscesses in children are located in the retropharyngeal or in the lateral pharyngeal space medial to the great vessels. Therefore, most can be managed successfully with intraoral rather than external drainage. External approaches are better reserved for those abscesses that are lateral to the great vessels or that involve multiple spaces. In this patient population, LPAs were more commonly seen than RPAs. PMID- 9413353 TI - Anterior cricoid split. Use of hyoid as autologous grafting material. AB - OBJECTIVE: To examine the use of hyoid as a readily available autologous grafting material for the anterior cricoid split (ACS) procedure. DESIGN: Prospective analysis of 20 patients undergoing ACS with hyoid interposition grafting for subglottic stenosis over a 3-year period. The patients received at least 1 year of follow-up after surgery. SETTING: Tertiary care children's hospital. PATIENTS: Twenty infants (age range, 2-9 months) with endoscopically confirmed acquired and congenital subglottic stenosis. Presenting symptoms included stridor, failure to extubate, and recurrent atypical croup. All 20 children underwent ACS with hyoid interposition grafting. RESULTS: All 20 patients exhibited improvement in their symptoms of airway obstruction. All 12 patients in whom extubation had previously failed subsequently underwent successful extubation. The 8 patients with symptoms of stridor and atypical croup showed marked improvement in their symptoms. Serial bronchoscopy revealed mucosal healing and incorporation of the hyoid grafts. CONCLUSION: Hyoid provides a readily available and reliable grafting material for interposition grafting in the ACS procedure for neonates and infants. PMID- 9413354 TI - alpha-difluoromethylornithine ototoxicity. Chemoprevention clinical trial results. AB - OBJECTIVES: To determine the effects of low-dose oral eflornithine hydrochloride (difluoromethylornithine [DFMO]) administration on hearing and to identify factors that influence those effects. DESIGN: Combined data from 2 studies: a prospective, randomized phase 1 clinical trial of eflornithine (n = 26 subjects) and a prospective, randomized, placebo-controlled phase 2 clinical trial of eflornithine (n = 40 subjects). SETTING: Ambulatory academic tertiary care referral center. PARTICIPANTS: Sixty-six volunteer subjects who had previously treated bladder, prostate, or colon cancer with no current evidence of neoplastic disease, or who were healthy individuals at increased risk for colon cancer, all without need of hearing amplification. INTERVENTIONS: Subjects were randomized to receive oral eflornithine at daily doses between 0.5 and 3 g per square meter of body surface area (g/m2) for 6 to 12 months (phase 1 study) or randomized to receive placebo or eflornithine, 0.5 g/m2 for 12 months (phase 2 study). OUTCOME MEASURES: Auditory thresholds were measured before, during, and after eflornithine administration. Auditory thresholds and threshold shifts were evaluated with regard to eflornithine dose, serologic variables, and demographic factors. RESULTS: Predictable shifts in auditory thresholds occurred following administration of eflornithine. As the daily dose of eflornithine increased, the magnitude and incidence of threshold shift increased, and the time until onset of threshold shift decreased. Threshold changes were greater in the lower frequencies than in the higher frequencies. Subjects' sex, age, and renal function had no effect on eflornithine-associated threshold shifts. Threshold shifts were reversible after eflornithine treatment was discontinued. CONCLUSIONS: Administration of eflornithine is associated with a predictable shift in auditory thresholds. The magnitude and incidence of threshold shift correlate with the daily eflornithine dose. PMID- 9413355 TI - Pseudomonas aeruginosa adherence to external auditory canal epithelium. AB - BACKGROUND: Pseudomonas aeruginosa is an important causative agent for external otitis. The specific bacterium-host reaction has not been investigated. It is therefore unknown whether adhesion of the external otitis strain to the external auditory canal epithelium is increased compared with strains isolated from other infections. DESIGN: A cohort study was designed to outline adhesion of P aeruginosa to the external auditory canal epithelium, cultured in vitro, of the guinea pig. Factors important for pathogenesis were also studied. PATIENTS: Pseudomonas aeruginosa strains from nonhospitalized patients were collected consecutively at the bacteriological laboratories at Karolinska Hospital, Stockholm, and Huddinge Hospital, Huddinge, Sweden. External otitis strains were compared with strains from leg ulcers, urinary tract infections, and cystic fibrosis. METHODS: Adhesion to the external auditory canal epithelium cultured in vitro was measured and compared groupwise with the mean profile of pathogenic factors. RESULTS: Adhesion to the epithelium was significantly increased for external otitis strains. These strains also had a significantly increased deoxyribonuclease production and a significantly decreased production of pyocyanin and alginate. CONCLUSIONS: The significantly increased ability of P aeruginosa, isolated from external otitis, to adhere to external auditory canal epithelium was combined with a significant production of pathogenic factors. The P aeruginosa that causes external otitis could therefore be considered a particular phenotype. The enzyme profile for external otitis strains was similar to that of the control groups except for the strains from cystic fibrosis. Adhesion to guinea pig vs human epithelium must be compared, and the effects of extracellular proteins on adhesion should be studied to further understand how P aeruginosa adheres to the external auditory canal. PMID- 9413356 TI - Significance of epidermoid formations in the middle ear in fetuses and children. AB - OBJECTIVE: To determine the incidence, size, and location of epidermoid formations (EFs), which have been suggested to be precursors of congenital cholesteatomas, in temporal bones from fetuses and children. DESIGN: We examined temporal bones from 226 fetuses and children up to the age of 10 years for the incidence, size, and location of EFs. RESULTS: Twenty-five EFs were identified in middle ears of 3 fetuses, 7 neonates, 9 infants, and 2 children aged 2 and 3 years. There was a male-female preponderance of 5:4. Generally, we saw EFs between the anterosuperior edge of the eardrum and the anterior limb of the tympanic ring, but 4 were below the level of the handle of the malleus. Their widths ranged from 25 to 300 microns. Keratinization was not observed in any EF. Contrary to previous reports, we found EFs not only in ears of fetuses, but also in ears of infants and children. CONCLUSION: Although EFs may persist in some ears, possibly developing into congenital cholesteatomas, our findings do not provide direct support for this concept. PMID- 9413357 TI - Safety of in vivo adenovirus-mediated thymidine kinase treatment of oral cancer. AB - BACKGROUND: Adenovirus-mediated transfer of the herpes simplex virus thymidine kinase gene (tk) is one of the most effective gene therapy strategies for solid tumors in experimental animal studies. Foundational animal studies in an oral cancer model have demonstrated significant antitumor effects and improved animal survival using this treatment strategy. OBJECTIVE: To assess the safety of adenovirus-mediated transfer of the herpes simplex virus tk gene for the treatment of oral cancer. DESIGN: Oral tumors were established in C3H/HeJ mice and were treated with tk followed by systemic ganciclovir administration. Polymerase chain reaction amplification techniques were used to screen local surrounding tissues and distant organs for the presence of the adenoviral construct. Microscopic examination of the tissues was performed to determine the cytopathic effects of the vector. Blood samples were obtained from the animals to test for liver, renal, and bone marrow function after treatment. RESULTS: The adenoviral vector was present in the livers, lungs, and kidneys of animals treated with the maximal single injection dose of 2 x 10(9) plaque forming units (pfu). No vector was noted systemically after delivery of an equally effective low dose of 1 x 10(8) pfu. Microscopic examination revealed no cytopathic effects in distant organs despite the presence of vector. Results of liver and renal function tests revealed no differences between treated and control animals. There was no statistical difference in white blood cell count, hematocrit, or platelet count between animals treated with ganciclovir and control animals. CONCLUSIONS: Based on these results, the direct delivery of adenovirus-tk followed by ganciclovir administration appears both efficacious and safe in an animal model. However, serum evaluation for adenovirus vector and screening organ function studies should be included in human protocols using this gene therapy scheme. PMID- 9413358 TI - Mandibular fracture and associated cervical spine fracture, a rare and predictable injury. Protocol for cervical spine evaluation and review of 1382 cases. AB - OBJECTIVES: To assess the relationship and incidence of cervical spine injuries in patients with mandibular fractures and to recommend an organized approach to cervical spine evaluation in these patients. DESIGN: A retrospective review of medical records of all patients with mandibular fractures at a level I trauma hospital from 1984 through 1993. Patient demographics, injury, mechanism of injury, associated symptoms, physical presentation, and adjuvant radiographic evaluations were recorded. SETTING: Level I, 1000-bed, urban trauma center in Atlanta, Ga. RESULTS: A total of 1382 patients with mandibular fractures were examined during the 10-year period of review. Cervical spine radiographs were obtained on 501 (36.3%) of these patients. From these radiographs, only 8 cervical spine fractures were found. All of the patients with cervical spine injuries (n = 8) had other associated maxillofacial injuries (n = 4), were involved in a motor vehicle accident (n = 7), or sustained gunshot wounds (n = 1). CONCLUSIONS: Judicious use of cervical spine radiographs in the appropriate setting of mandibular trauma is beneficial. However, clinical criteria should dictate rational use of radiographs, since the association between cervical spine injuries and mandibular trauma is rare and predictable. PMID- 9413359 TI - The zygomatic-sphenoid fracture line in malar reduction. A cadaver study. AB - OBJECTIVE: To demonstrate the persistent malar displacement and distraction of the zygomatic-sphenoid fracture line that is possible after alignment of the frontozygomatic and infraorbital rim fractures of a displaced malar fracture. DESIGN: Nonblinded cadaver study. SUBJECTS: Three fresh cadaver heads and 1 representative clinical example. INTERVENTION: The cadaver heads were subjected to blunt trauma to the malar eminence. Reduction of the malar unit was performed either with attention to the frontozygomatic and infraorbital rim fractures alone or with concomitant inspection of the zygomatic-sphenoid fracture line. The representative case was repaired with a trans-conjunctival approach for inspection of the zygomatic-sphenoid fracture line. RESULTS: Persistent malar asymmetry is possible after the reduction of displaced malar fractures when only the frontozygomatic and infraorbital rim buttresses are used for reference. In each case in our study, the zygomatic-sphenoid fracture line remained distracted. Alignment of the zygomatic-sphenoid fracture restored premorbid malar position. CONCLUSION: Inspection of the zygomatic-sphenoid fracture line can contribute significantly to the precise 3-dimensional reduction of displaced malar fractures. PMID- 9413360 TI - Craniofacial resection for malignant tumors involving the anterior skull base. AB - OBJECTIVES: To review our experience with craniofacial resection for malignant neoplasms of the anterior skull base and report long-term results, and to analyze survival in terms of the overall experience, tumor histological diagnoses, and tumor extent. Also, to report complications of this surgical procedure. DESIGN: Retrospective review. SETTING: Tertiary cancer facility. PATIENTS: We evaluated 115 consecutive patients undergoing craniofacial resection for malignant neoplasms involving the anterior skull base. Forty-five (39%) presented with recurrent or persistent disease after prior therapy. MAIN OUTCOME MEASURES: Survival was evaluated with the Kaplan-Meier product limit method and comparisons between individual subgroups were performed using the log-rank test. RESULTS: The operative mortality rate was 3.5%. Major complications occurred in 40 patients (35%). For the entire group, disease-specific survival rates were 58% and 48% at 5 and 10 years, respectively. The highest survival rate was observed in patients with esthesioneuroblastoma and lowest in those with mucosal melanoma. Survival was significantly better for those whose tumors could be excised with a limited resection in comparison with those requiring an extended procedure (P = .009). CONCLUSIONS: A 23-year experience with craniofacial resection performed for malignant tumors involving the anterior skull base confirms the durable results obtained with this intervention. The diversity of histological diagnoses, site of origin, extent of tumor invasion, and impact of prior therapy hampers any attempt at reporting meaningful survival statistics for comparison with other series or other means of treatment. PMID- 9413361 TI - Extranodal lymphomas of the head and neck. A 20-year experience. AB - BACKGROUND: Extranodal non-Hodgkin lymphoma (NHL) of the head and neck is a relatively uncommon disease. Over the last 3 decades, a variety of systems, including the Rappaport, Luke-Collins, and Working Formulation classifications, have been used to classify extranodal NHLs of the head and neck. Most studies have included a relatively small number of patients, used different modalities of therapy, and did not include all head and neck sites. These limitations make comparisons between different studies and drawing any conclusions difficult. OBJECTIVES: To describe in a uniform fashion a relatively large number of patients with extranodal NHL of the head and neck treated at the same institution, using only the most current classification system and to describe the clinical features, behavior, and outcome of this relatively uncommon, but potentially curable disease. DESIGN: A retrospective study of 98 patients with extranodal NHL of the head and neck. All patients were reclassified according to the Working Formulation system (regardless of the time of diagnosis) in order to uniformly define the clinical course of this disease in the head and neck. SETTING: A tertiary care referral center. RESULTS AND CONCLUSIONS: The sinonasal tract was the most commonly involved site (25%). If the nasopharynx (16%), tonsil (12%), and base of tongue (8%) are grouped together, this combined site (Waldeyer ring) becomes the most common site of disease (36%). Patients with tonsillar lymphoma had a 20% incidence of associated gastrointestinal involvement. Approximately 50% of the patients had associated nodal disease, and only 20% had systemic or B symptoms. Three fourths of the patients had stage I or II disease, and approximately two thirds had intermediate-grade lymphoma. Radiation therapy was the primary modality of therapy for localized disease (stages I and II), especially for low-grade lymphomas. Combination chemotherapy with or without radiation was used for more advanced disease and for intermediate- and high-grade lymphomas. Surgery was limited to establishing the diagnosis. Two thirds of the patients had a remission after initial therapy. Two thirds of these patients had no further relapse. Three fourths of the patients with relapse after initial remission died of their disease. The overall and disease-free survival rates for all patients were 60% and 50%, respectively. Outcome of therapy was related to stage and histologic grade. Patients with lymphomas of high histopathologic grade and recurrent and recurrent and disseminated disease had the poorest prognosis. PMID- 9413362 TI - Versatility of the free anterolateral thigh flap for reconstruction of head and neck defects. AB - OBJECTIVE: The anterolateral thigh flap has many advantages in head and neck reconstruction. However, it has not yet come into widespread use because of the anatomic variations of its perforators. Herein, we describe a safe operative technique related to the patterns of the perforators and discuss its wide versatility. SETTING: A national cancer center hospital. PATIENTS: Thirty-eight anterolateral thigh flaps were transferred. Confirmation and dissection of the flap pedicle were simultaneously performed with tumor resection. The design and elevation of the flap were carried out immediately after the tumor resection was completed. RESULTS: From the study of the anatomic variations of the perforators, septocutaneous patterns were recognized in 10 cases (26.3%) and musculocutaneous patterns in 28 cases (73.7%). All flaps were easily and safely elevated with our techniques. Thirty-six flaps survived. Partial necrosis was noted owing to excessive thinning procedure in one patient and total necrosis was noted owing to venous thrombosis at the anastomosis part in another patient. CONCLUSIONS: We found that the anterolateral thigh flap has numerous advantages. It is possible to perform the flap elevation and the tumor resection simultaneously. The flap is generally thin and is suitable for reconstruction of intraoral defects. Combined flaps with neighboring tissues and other, distant flaps can be used. Furthermore, since our technique minimizes the problems of confirmation and dissection of the perforators, we conclude that this flap can be successfully used to repair a variety of large defects of the head and neck. PMID- 9413363 TI - Overcoming the learning curve in microvascular head and neck reconstruction. AB - BACKGROUND: It is widely accepted that most microvascular reconstructive surgeons experience a learning curve. A compilation of 6 series of microvascular surgery reported in the literature revealed that the average rate of successful free flap transfer rose from 79% to 96% as the surgeons gained clinical experience. OBJECTIVE: To review the collective experience of 3 otolaryngologist-head and neck surgeons performing free flaps during their first year of clinical practice after completion of postgraduate training. DESIGN: A multi-institutional retrospective case series. SETTING: Three academic tertiary care otolaryngology head and neck surgery programs. PATIENTS: Eighty-one microvascular free flaps were performed in patients undergoing surgical reconstruction of head and neck defects during a 1-year period. INTERVENTIONS: Free flap selection was based on specific defect characteristics. Radial forearm, fibula, and rectus abdominis flaps together accounted for 90% of the donor sites selected. MAIN OUTCOME MEASURE: Reported incidence of partial or complete free flap necrosis. RESULTS: There were 2 perioperative deaths. Among the surviving patients, there were 2 cases of complete flap failure, for an overall success rate of 97.5%. There were 2 additional cases of partial flap necrosis (2.5%) that were related to errors in flap insetting. CONCLUSION: The availability of high-quality postgraduate training combined with the judicious selection of free flaps that offer long vascular pedicles and large diameter vessels can allow junior microvascular head and neck surgeons to achieve free flap survival rates that are comparable with those reported by experienced microvascular surgeons. PMID- 9413365 TI - Complications following airway surgery in Noonan syndrome. AB - Two children with Noonan syndrome underwent airway surgery and both experienced spontaneous chylothorax after surgery. The Noonan syndrome phenotype may include defects that significantly increase the risk of complications associated with surgery. Manifestations of Noonan syndrome may include congenital heart disease; coagulation factor deficiency; pterygium colli; and lymphangiomatosis of the pleura, lungs, and chest wall that can lead to life-threatening chylothorax. In this article, the 2 cases are presented and the relevant literature is reviewed to increase awareness of the potential problems that may be encountered in patients with Noonan syndrome. Recommendations are made regarding preoperative investigations and perioperative management to prevent complications. PMID- 9413364 TI - Maxillary sinus mucosal blood flow during nasal vs tracheal respiration. AB - OBJECTIVE: To determine the effect of changes from nasal to tracheal respiration on maxillary sinus mucosal blood flow in rabbits with unobstructed sinus ostia. DESIGN: Animals underwent tracheotomy with a T tube and then a small window of intact maxillary sinus mucosa was exposed. Mucosal blood flow was recorded during normal nasal respiration using laser-Doppler velocimetry. At hourly intervals, respiration was changed from the nasal to the tracheal route and then back again. SUBJECTS: Ten anesthetized rabbits were used: 5 underwent single and 4 underwent multiple shifts in the respiratory route, while 1 was monitored continuously during long-term nasal breathing only. RESULTS: A significant decrease in maxillary sinus blood flow occurred on switching from nasal to tracheal respiration and a significant increase in blood flow occurred on return to nasal respiration. Where multiple switches were made, blood flow changes diminished in magnitude, but significant decreases (nasal to tracheal) or increases (tracheal to nasal) were evident in all cases. CONCLUSIONS: It is proposed that the maxillary sinus may act in an accessory capacity to the nose for humidification of inspired air via secretions liberated from the sinus ostium. Furthermore, we suggest that nasal airflow is involved with the reflex regulation of sinus blood flow, probably via stimulation of sensory receptors in the nasal cavity. Reduced maxillary sinus mucosal blood flow may thus contribute to supra-systemic levels of antral carbon dioxide. Since elevated carbon dioxide levels have been shown to reduce maxillary sinus mucociliary activity in vitro, nasal airflow compromise may contribute to the initiation of a cascade of pathophysiological events leading to acute sinusitis. PMID- 9413366 TI - The bridging bronchus. Successful diagnosis and repair. AB - Anomalies of bronchial branching are infrequent and may be difficult to diagnose. The bridging bronchus is a rarely reported anomaly that may not be as sporadic as once thought. We describe an infant with respiratory distress whose right middle and lower lobes were supplied by a bridging bronchus arising from the left main bronchus. The airway anatomy was defined using flexible and rigid bronchoscopy and helical computed tomographic scanning, enabling successful surgical repair. We review current literature on the bridging bronchus as well as the possible embryological basis for this defect. PMID- 9413367 TI - Obstructive sleep apnea in Schwartz-Jampel syndrome. AB - Schwartz-Jampel syndrome (SJS) is a rare entity characterized by myotonia and skeletal abnormalities. Death and respiratory distress have previously been reported in newborns and young children with SJS. We describe a patient with SJS and snoring in whom polysomnography demonstrated obstructive sleep apnea and hypoxia. Although tonsillectomy with laser palatoplasty significantly widened the oropharyngeal introitus, obstructive sleep apnea persisted. Ultimate improvement occurred only after the institution of home therapy with bi-level positive airway pressure during the night. We also discuss the specific structural and neuromuscular features of SJS that may be responsible for upper airway obstruction. PMID- 9413368 TI - Pathologic quiz case 1. Adenoid cystic carcinoma of the parotid gland. PMID- 9413369 TI - Pathologic quiz case 2. High-grade osteosarcoma of the maxilla. PMID- 9413370 TI - Duplication of internal jugular vein. PMID- 9413371 TI - Carotid endarterectomy--what are we doing? PMID- 9413372 TI - New insights into inflammatory abdominal aortic aneurysms. PMID- 9413373 TI - Vein grafts: haemodynamic forces on the endothelium--a review. AB - OBJECTIVE: To review the mechanisms believed to be important in the development of vein graft stenosis, with particular attention placed on the adaptation of saphenous vein endothelium to a new haemodynamic environment. DESIGN AND METHODS: Discussion based on review of published research. RESULTS: The aetiology of vein graft stenosis remains to be established and appears to be multi-factorial. The increasing evidence for an important role of haemodynamic forces is discussed, particularly via the interaction of these force with the endothelium. CONCLUSION: Further understanding of the interaction between haemodynamic forces, blood constituents and the newly implanted vein graft is required. Use of in vitro models is contributing increasing knowledge to this area, but ultimately better non-invasive methods of assessing haemodynamic forces in vivo are required. PMID- 9413374 TI - Exercise training for intermittent claudication: does it adversely affect biochemical markers of the exercise-induced inflammatory response? AB - OBJECTIVES: To identify a stable biochemical marker of disease severity in patients with intermittent claudication and to use these findings to assess the effect of therapeutic exercise training. DESIGN: Case-control study: prospective randomised-controlled trial of exercise training. MATERIALS AND METHODS: Plasma fibrinogen, serum amyloid A protein (SAA), C-reactive protein (CRP) and urinary albumin-creatinine ratio (ACR) were measured in 67 claudicants and 15 controls. Twenty-two patients were randomised to supervised exercise training and 17 randomised to observation. Subjects were reviewed at 3, 6 and 12 months. RESULTS: The median (interquartile range) baseline fibrinogen was 3.7 g/l (3.3-4.25) in claudicants and 3.5 g/l (2.9-3.95) in controls (p = 0.045); CRP was 4.7 mg/l (2.2 9.0) and 2.1 mg/l (1.0-2.8), respectively (p < 0.0001); SAA was 72 mg/l (35-132) and 30 mg/l (20-89) (p = 0.0009). Claudicants showed an increased urinary ACR following treadmill exercise (Wilcoxon, p < 0.0001) with no change in controls. Exercise training reduced SAA at 6 months, CRP at 3 months and progressively attenuated the post-exercise increase in ACR. No similar changes were found in controls. CONCLUSIONS: Repetitive low-grade inflammatory events in claudicants lead to elevation of serum acute-phase proteins. Exercise training is associated with symptomatic improvement and reduction inflammatory markers. The concern that exercise has adverse systemic effects therefore seems to be unjustified. PMID- 9413375 TI - The inflammatory response to vascular surgery-associated ischaemia and reperfusion in man: effect on postoperative pulmonary function. AB - OBJECTIVES: To characterise the inflammatory response to vascular surgery and ischaemia/reperfusion (I/R) in man, regarding release of inflammatory mediators, recruitment and activation of neutrophils, and their relation to postoperative pulmonary function. DESIGN: Prospective cohort study. MATERIALS AND METHODS: Circulating neutrophil counts and plasma levels of elastase-alpha 1-antitrypsin (AT), a neutrophil degranulation product, were measured before and approx. 2.5 h (group 1, n = 19) after elective abdominal aortic surgery, and approx. 2.9 h after elective peripheral vascular surgery (group 2, n = 6), together with concentrations of neutrophil agonists, including activated complement (C3a), secretory phospholipase A2 (sPLA2), tumor necrosis factor (TNF-alpha), interleukin (IL)-6, IL-8 and granulocyte colony-stimulating factor (G-CSF). At the time of blood sampling, respiratory variables allowing computation of the lung injury score (LIS) were obtained in patients admitted after surgery in the intensive care unit (ICU), i.e. all group 1 patients and one group 2 patient. RESULTS: Median (range) neutrophil counts rose by 80% (-28-208) and 90% (10-147) in groups 1 and 2, respectively (n.s. between groups). The increase (p < 0.05) in elastase-alpha 1-AT level was 121% (-5-439) in group 1 and 82% (18-792) in group 2 (n.s. between groups). There was a rise (p < 0.05) in C3a level by 93% (-42 751) and of sPLA2 level by 68% (-40-1400) after surgery for the groups together (n.s. between groups), and the rise of the elastase-alpha 1-AT related to that of the C3a levels. IL-6 and G-CSF concentrations increased more in group 1 than 2. The IL-8 concentration increased in group 1 only, and TNF-alpha was unchanged in all groups. In ICU patients, the LIS related to the postoperative rise in IL-6 level only, even though the rise in plasma concentrations of cytokines interrelated. No patient developed ARDS and all survived. CONCLUSIONS: Vascular surgery and I/R in man activates complement, releases cytokines (except for TNF alpha), and induces neutrophil recruitment and degranulation, which may primarily depend on complement activation. In contrast to the latter, the release of cytokines may depend on the extent of I/R and may contribute to transient pulmonary dysfunction after extensive I/R. PMID- 9413376 TI - Retrograde aortic and selective organ perfusion during thoracoabdominal aortic aneurysm repair. AB - OBJECTIVES: To evaluate the possible prevention of renal and intestinal ischaemia during surgery of thoracoabdominal aortic aneurysms (TAAA) by use of retrograde and selective organ perfusion. DESIGN: Prospective study. MATERIALS: Thirty-three consecutive patients underwent TAAA repair, six of whom had a previous type B dissection: 14 patients (35%) had type I TAAA, 12 patients type II (32%), three patients type III (15%) and four patients type IV (18%). Mean age was 61 years (range 22-84 years). METHODS: In patients with type I TAAA, retrograde aortic perfusion was performed by means of a left atrium femoral artery bypass or partial cardiopulmonary bypass. In type II, III and IV the same procedure was performed; however, following cross-clamping and opening of the abdominal aorta, the coeliac trunc, superior mesenteric and both renal arteries were selectively perfused with four Pruitt-catheters (9 Fr.), connected as an octopus to the extracorporal circulation. RESULTS: All patients survived the surgical procedure. The minimal volume flow through each octopus catheter was 60 ml/min. Urine output was uninterrupted in all patients, irrespective of the aortic cross-clamp time. Only one patient (3%), who already had renal insufficiency, developed renal failure. Total in-hospital mortality was 15%, paraplegia occurred in 12%. CONCLUSION: Retrograde aortic and selective organ perfusion is a safe technique and can prevent ischaemic renal and intestinal damage during cross-clamping of the aorta in thoracoabdominal aneurysm surgery. PMID- 9413377 TI - Venous thrombectomy for iliofemoral vein thrombosis--10-year results of a prospective randomised study. AB - OBJECTIVES: To study if venous thrombectomy prevents late post-thrombotic sequelae, venous obstruction reflux, and improves venous physiology following an acute iliofemoral venous thrombosis. DESIGN: Prospective randomised controlled study. MATERIAL: Thirty patients returned for follow-up 10 years after an acute iliofemoral venous thrombosis initially treated with conventional anticoagulation treatment (medical group, n = 17) or with thrombectomy combined with a temporary arteriovenous fistula and anticoagulation (surgical group, n = 13). Clinical assessment, radionuclide angiography, duplex ultrasound and venous physiology tests were performed. RESULTS: Leg swelling was recorded in 12 (71%) and leg ulcers in three (18%) of the medical patients and in, respectively, six (46%) and one (8%) of the surgical patients. The surgical patients had less severe sequelae (class 0-2). Radionuclide angiography demonstrated that the iliac vein was more commonly occluded following medical (59%) than following surgical (17%) treatment (p < 0.05). Duplex examination demonstrated slightly (n.s.) more reflux in the femoral and popliteal veins in the medical group. Venous physiology (occlusion plethysmography, foot volumetry, and foot vein pressures) did not show any significant differences, although the medical group tended to have a more severe pathology. CONCLUSION: Venous thrombectomy improves venous patency and possibly reduces venous reflux and post-thrombotic sequelae as compared to anticoagulation treatment. PMID- 9413378 TI - Mortality in abdominal aortic aneurysm surgery--the effect of hospital volume, patient mix and surgeon's case load. AB - OBJECTIVE: Assessment of mortality in abdominal aortic aneurysm surgery. DESIGN: A 4-year cross sectional study based on a nationwide vascular registry: Finnvasc. MATERIAL AND METHODS: A total of 17,465 vascular interventions included 929 elective repairs for abdominal aortic aneurysms (AAA), and 610 emergency cases with 454 ruptures. Fifty-three percent of the operations were done in university hospitals, 44% in central hospitals and 3% in district hospitals. RESULTS: The 30 day mortality rate for AAA repair was 5.1% in elective and 46% in ruptured cases. A clear dependence of operative mortality on surgeon's experience in AAA surgery was observed, both regarding the surgeon's total vascular case load (p < 0.01) and the number of operated elective aneurysms (p < 0.01), but not the number of operated ruptured aneurysms. However, no association was found between hospital volume and mortality in AAA surgery. CONCLUSIONS: Vascular surgical experience clearly improves the results of elective aneurysm surgery. PMID- 9413379 TI - Percutaneous angioplasty for infrainguinal graft-related stenoses. AB - OBJECTIVE: To assess the success of percutaneous transluminal angioplasty (PTA) in treating infrainguinal graft-related stenoses. DESIGN: Retrospective analysis of stenoses undergoing PTA over 6 years. MATERIALS: Fifty-seven stenoses in 42 grafts. METHODS: Site, length and type of stenoses recorded. Follow-up till discharge, graft occlusion or death. RESULTS: PTA was successful in 48/57 stenoses in 36 grafts (G), with a poor result in seven. Further PTA was required in seven stenoses (7 G). One graft occluded at PTA and one stenosis was inaccessible. Overall graft (G) patency (median 13 months) was 82% (1 year patency 84%). Of 48 successful PTAs (37 G), 36 remained patent (28 G), eight (4 G) occluded and four were lost to follow-up (4 G). Fourteen of thirty-six stenoses which remained patent required further intervention (seven PTA, six jump grafts, one vein patch). The four occlusions were associated with small veins (two), multiple stenoses (one) and a PTFE graft which occluded 10 days following PTA. Of the seven PTAs with a poor angiographic result, five remained patent, three after further intervention. CONCLUSION: PTA is the best treatment for localised stenoses. Stenoses > 2 cm or multiple (three or more) stenoses are best treated surgically. Follow-up is essential, as 20% require further intervention. PMID- 9413380 TI - Carotid endarterectomy in the U.K. and Ireland: audit of 30-day outcome. The Audit Committee for the Vascular Surgical Society. AB - OBJECTIVES AND DESIGN: A prospective study of 709 patients undergoing carotid surgery in the U.K. and Ireland was performed to evaluate the performance of vascular surgeons. MATERIALS AND METHODS: Fifty-nine surgeons (range 2-39 cases each) were sampled and all patients undergoing surgery over a 6-month period (1 March 1994-31 August 1994) were included in the study. Indications for surgery were TIA (35.9%), AF (23.3%), CVA (21.4%) and "others" (19.6%). RESULTS: Mean ipsilateral stenosis was 82% (30%-99%). Thirty-one percent of patients had preoperative neurological consults. Shunts were used in 67.6%, tacking sutures in 40.1%, drains in 71.9% and patches in 54.4% of cases. At 30 days there were nine (1.3%) deaths (four cardiac, three neurological). There were 15 ipsilateral postoperative CVAs (2.1%); 19% of patients had one or more complication, usually minor. Statistical analysis showed no independent risk factor for CVA other than seniority of the surgeon. CONCLUSIONS: A combined stroke/death rate of 3% for the series was obtained at 30 days for all cases. This large, validated study suggests that members of the Vascular Society of G.B. and Ireland currently have a very low morbidity/mortality rate for performing carotid surgery. Continued audit is required to ensure that this quality of service does not deteriorate. PMID- 9413381 TI - Haemostatic and rheological factors as predictors of restenosis following percutaneous transluminal angioplasty. AB - OBJECTIVES: To determine whether pre-angioplasty levels of haemostatic and rheological factors predicted restenosis of the dilated arterial segment following percutaneous transluminal angioplasty. DESIGN AND SETTING: Prospective study, Two regional hospital centres for angioplasty in Edinburgh and Glasgow, Scotland, UK. METHOD: Haemostatic and rheological factors were measured in 102 subjects with atherosclerotic disease of the lower limbs, immediately prior to percutaneous transluminal angioplasty. Subjects were followed up after 2-3 years for restenosis of the original angioplasty site using duplex scanning. RESULTS: Baseline clinical characteristics were similar between subjects who restenosed (n = 27) and those with no restenosis (n = 39), except that occluded lesions were more likely to restenose than stenoses (p < or = 0.05). There was no significant difference in age- and sex-adjusted mean levels of whole blood viscosity, plasma viscosity, haematocrit, von Willebrand factor, fibrin D-dimer or plasminogen activator inhibitor-1 activity between the stenosed and no restenosis groups (p > 0.1), but mean plasma fibrinogen was lower in the restenosed group (3.31 g/l vs. 3.75 g/l; p < or = 0.05). These results persisted after multivariate adjustment for smoking habit and type of lesion dilated. CONCLUSIONS: This study provides no evidence that raised, pre-angioplasty levels of haemostatic and rheological factors predict restenosis following percutaneous transluminal angioplasty of the arteries of the lower limbs. PMID- 9413382 TI - Middle cerebral artery blood velocity, embolisation, and neurological outcome during carotid endarterectomy: a prospective comparison of the Javid and the Pruitt-Inahara shunts. AB - OBJECTIVES: To investigate the in vivo haemodynamic performance and neurological outcome of two types of carotid shunt. DESIGN: Randomised single surgeon study of consecutive symptomatic patients. SETTING: 163 consecutive patients undergoing carotid endarterectomy for symptomatic carotid disease were randomised to the Javid or Pruitt shunt. CHIEF OUTCOME MEASURES: Middle cerebral artery velocity (MCAV), preoperatively, during clamping, during shunting and post-restoration of flow, embolic episodes, neurological outcome. MAIN RESULTS: The MCAV preoperatively, at carotid clamping, and postoperatively was the same for both groups (p > 0.15). During shunting the MCAV was significantly lower in the Pruitt group, p < 0.005, 59% of the Javid and 34% of the Pruitt shunts maintained MCAV at preoperative levels p < 0.005, chi 2 = 8.92. The Javid shunt produced significantly more emboli (73% of cases) at declamping than the Pruitt (41%), p < 0.0002, chi 2 = 14.7. Four Javid patients and one Pruitt had disabling thromboembolic strokes; overall thromboembolic stroke rate 3.7%. The difference in stroke rates was not statistically significant (p = 0.14). CONCLUSIONS: The Pruitt shunt was unable to maintain preoperative MCAV in 66% of cases, the Javid shunt had a higher incidence of emboli on declamping. These factors may lead to an increased risk of stroke; however, the numbers required for statistical confirmation would be large. PMID- 9413383 TI - Recurrent coronary-subclavian steal syndrome treated by left subclavian artery stenting. PMID- 9413384 TI - Abdominal aortic aneurysm associated with horseshoe kidney and duplication of inferior vena cava--an extra-peritoneal approach. PMID- 9413385 TI - Spontaneous rupture of the abdominal aorta without pre-existing aneurysm--two case reports. PMID- 9413386 TI - Motor cycling and finger ischaemia. PMID- 9413387 TI - Left gastric artery aneurysm--a case report. PMID- 9413388 TI - Limb volume. PMID- 9413389 TI - Carotid surgery trials. PMID- 9413390 TI - Carotid surgery and ocular ischaemia. PMID- 9413391 TI - Graft failure and amputation. PMID- 9413392 TI - Adhesion and signaling in vascular cell-cell interactions. PMID- 9413393 TI - Endothelial adherens junctions: implications in the control of vascular permeability and angiogenesis. PMID- 9413394 TI - Genetic manipulation of vascular adhesion molecules in mice. PMID- 9413395 TI - The extracellular matrix as a cell cycle control element in atherosclerosis and restenosis. PMID- 9413396 TI - Effects of fluid dynamic forces on vascular cell adhesion. PMID- 9413397 TI - The biology of PECAM-1. PMID- 9413398 TI - Selectin ligands: will the real ones please stand up? PMID- 9413399 TI - Cell adhesion and angiogenesis. PMID- 9413400 TI - von Willebrand factor. PMID- 9413401 TI - Therapeutic inhibition of carbohydrate-protein interactions in vivo. PMID- 9413402 TI - Integrins and vascular extracellular matrix assembly. PMID- 9413403 TI - Platelet GPIIb/IIIa antagonists: the first anti-integrin receptor therapeutics. PMID- 9413405 TI - Heparan sulfate proteoglycans of the cardiovascular system. Specific structures emerge but how is synthesis regulated? PMID- 9413406 TI - New insights into integrin-ligand interaction. PMID- 9413404 TI - Biomechanical activation: an emerging paradigm in endothelial adhesion biology. PMID- 9413407 TI - Adhesive interactions of sickle erythrocytes with endothelium. PMID- 9413408 TI - Smooth muscle migration in atherosclerosis and restenosis. PMID- 9413409 TI - Integrin signaling in vascular biology. PMID- 9413410 TI - Role of PSGL-1 binding to selectins in leukocyte recruitment. PMID- 9413411 TI - Sex therapy in psychiatric treatment has a new partner: reproductive hormones. PMID- 9413412 TI - Comparative prophylactic efficacy of lithium, carbamazepine, and the combination in bipolar disorder. AB - BACKGROUND: We compared the prophylactic efficacy of lithium, carbamazepine, and the combination and identified possible clinical markers of response. METHOD: Fifty-two outpatients who met DSM-III-R criteria for bipolar illness were randomly assigned in a double-blind design for an intended 1 year of treatment with lithium or carbamazepine, a crossover to the opposite drug in the second year, and then a third year on the combination. Patients received monthly detailed evaluations, and daily life chart ratings of the degree of functional incapacity associated with mania or depression were completed. RESULTS: For evaluable patients: 13 (31.0%) of 42 failed to complete a full year of lithium therapy owing to lack of efficacy, and 2 dropped out because of side effects; 13 (37.1%) of 35 withdrew from carbamazepine within the first year owing to lack of efficacy, and 10 dropped out because of side effects (9 of the 10 had a rash); 7 (24.1%) of 29 withdrew from the combination therapy owing to lack of efficacy. The percentage of the evaluable patients who had marked or moderate improvement on the Clinical Global Impressions scale was 33.3% on lithium. 31.4% on carbamazepine, and 55.2% on the combination treatment, which was not significantly different. By a variety of measures, lithium was more effective than carbamazepine in the prophylaxis of mania. Patients with a past history of rapid cycling did poorly on monotherapy (28.0% responded to lithium; 19.0% responded to carbamazepine), but significantly better on the combination (56.3%, p < .05). CONCLUSION: These prospective, randomized data suggest a high incidence of inadequate response to either mood stabilizer or their combination despite use of adjunctive agents as needed. Additional novel treatment regimens are needed to better decrease affective morbidity in large numbers of bipolar outpatients. PMID- 9413413 TI - Olanzapine in treatment-refractory schizophrenia: results of an open-label study. The Spanish Group for the Study of Olanzapine in Treatment-Refractory Schizophrenia. AB - BACKGROUND: Clozapine is currently the treatment of choice for neuroleptic resistant schizophrenia. Olanzapine is a new antipsychotic drug that has shown efficacy against positive and negative symptoms of schizophrenia, with minimal extrapyramidal side effects. However, the effectiveness of olanzapine has not yet been reported among treatment-refractory schizophrenic patients. METHOD: A total of 25 schizophrenic patients (DSM-IV criteria) with documented lack of response to two conventional antipsychotic drugs entered this 6-week prospective, open label treatment trial with olanzapine 15 to 25 mg/day. An optional extension up to 6 months was provided. RESULTS: As a group, the olanzapine-treated patients showed statistically significant improvement (p < .05) in both positive and negative symptoms by the end of 6 weeks of therapy. Overall, 9 of the patients (36%) met the a priori criteria for treatment-response (> or = 35% decrease in Brief Psychiatric Rating Scale [BPRS] total score, plus posttreatment Clinical Global Impression-Severity < or = 3 or BPRS total < 18). Only one patient discontinued treatment because of an adverse event during the study. Despite the relatively high dosages of olanzapine used, there were no reports of parkinsonism, akathisia, or dystonia, and no patients required anticholinergic medication. CONCLUSION: This open study suggests that olanzapine may be effective and well tolerated for a substantial number of neuroleptic-resistant schizophrenic patients. Further blinded, controlled trials are needed to confirm our results. PMID- 9413415 TI - Delusional jealousy in dementia. AB - BACKGROUND: Delusional jealousy is a major motive for violence and is sometimes found in demented patients. This study was undertaken to investigate the frequency and the characteristics of delusional jealousy in demented patients. METHOD: The sample population consisted of 133 demented patients admitted to the geropsychiatric ward. Patients with and without delusional jealousy were compared in terms of general characteristics and psychotic symptoms. RESULTS: Of the 133 demented patients, 21 (15.8%) showed delusional jealousy. Delusional jealousy was found in various types of dementia. There were no significant differences between the two groups in regard to age, age at onset, gender, educational level, and Mini-Mental State Examination score. All patients with delusional jealousy had at least one other psychotic symptom. CONCLUSION: Delusional jealousy is a common problem in dementia. The psychobiological factors of delusional jealousy and cognitive function in demented patients may differ. There may be various underlying factors for the development of delusional jealousy in dementia. PMID- 9413414 TI - A double-blind, placebo-controlled study comparing the effects of sertraline versus amitriptyline in the treatment of major depression. AB - BACKGROUND: This study was designed to compare the efficacy, safety, tolerability profiles, and effects on quality of life of the serotonin selective reuptake inhibitor antidepressant sertraline versus the nonselective tricyclic antidepressant amitriptyline and placebo in patients with major depression. METHOD: Outpatients with DSM-III-R major depression were randomly assigned to double-blind treatment for 8 weeks with sertraline (50-200 mg daily), amitriptyline (50-150 mg daily), or matching placebo. Assessments included the Hamilton Rating Scale for Depression, Montgomery-Asberg Depression Rating Scale, Clinical Global Impressions-Severity of Illness scale, Clinical Global Impressions-Improvement scale, Global Assessment Scale, Profile of Mood States, Beck Depression Inventory, Quality of Life Enjoyment and Satisfaction Questionnaire, and Health-Related Quality of Life battery. RESULTS: All treatment groups demonstrated statistically significant improvement from baseline in depression ratings by Week 1 and thereafter. The antidepressant effects of amitriptyline and sertraline were significantly (p < .05) greater than placebo and did not differ significantly from each other. Sertraline was associated with significantly (p < .05) greater subjective (i.e., patient-rated) improvement in mood than amitriptyline or placebo. Both active drugs were associated with greater improvements than placebo on most quality of life measurements. On several items, sertraline, but not amitriptyline, was superior to placebo. There was a discernible effect of sertraline earlier than amitriptyline on most quality of life scales. Amitriptyline therapy was associated with significantly more treatment-related adverse events, and discontinuations due to treatment-related adverse events, in comparison to both sertraline and placebo therapy. CONCLUSION: Sertraline and amitriptyline each were effective treatments for major depression as assessed by both physician- and patient-rated scales. These results show that sertraline therapy is better tolerated than amitriptyline therapy. Quality of life was also improved by effective antidepressant treatment, with sertraline showing a tendency to produce greater improvements on quality of life measures. PMID- 9413416 TI - Severe atypical symptoms without depression in SAD: effects of bright light therapy. PMID- 9413417 TI - Is divalproex a cost-effective alternative in the acute and prophylactic treatment of bipolar I disorder? PMID- 9413418 TI - Resolution of fluoxetine-induced sexual dysfunction with the 5-HT3 antagonist granisetron. PMID- 9413419 TI - Olfactory reference syndrome responds to clomipramine but not fluoxetine: a case report. PMID- 9413420 TI - Affective illness and epilepsy. PMID- 9413421 TI - Blood clozapine levels elevated by fluvoxamine: potential for side effects and lower clozapine dosage. PMID- 9413422 TI - The pharmacologic rationale for the clinical use of antidepressants. PMID- 9413423 TI - Determinants of suicidal ideation: the role of substance use disorders. AB - BACKGROUND: This study tested the hypothesis that the amount of psychoactive substance consumed (frequency and/or quantity), life problems resulting from this use, and a DSM-IV diagnosis of substance abuse/dependence are independent risk factors associated with increased suicidal ideation in a population of psychiatric inpatients with major depressive disorder. METHOD: 891 hospitalized patients with a primary diagnosis of nonpsychotic major depressive disorder (MDD) received a standardized, psychiatrist-administered assessment battery. To examine the relationship between admission suicidality and demographic, psychiatric history, and admission variables, chi-square analyses were used for categorical data and one-way ANOVAs were used for continuous indices. Stepwise hierarchical multiple regression analyses were performed to determine the set of variables that was independently related to admission suicidality level. RESULTS: There was general agreement between our findings and previous literature in regard to the association between severity of Axis I diagnosis, depressed mood, hopelessness, male gender, unemployment, involuntary treatment, and alcohol/drug problems and higher suicidal ideation. In our sample of hospitalized patients with unipolar major depressive disorder, higher current drug and/or alcohol dependency and high current use of alcohol or other substances of abuse were independently associated with higher levels of suicidal ideation. CONCLUSION: This association with higher suicidal ideation lends support to the importance of treating patients for both alcohol/drug problems and depression in an effort to decrease their risk for future suicide. We hope that our findings will improve the care that patients with dual diagnoses receive. PMID- 9413424 TI - von Hippel-Lindau disease. AB - von Hippel-Lindau disease is a hereditary cancer syndrome characterized by the development of vascular tumors of the central nervous system and retina, clear cell renal carcinomas, pheochromocytomas, pancreatic islet cell tumors, endolymphatic sac tumors, and benign cysts affecting a variety of organs. VHL disease is caused by germline mutations of the von Hippel-Lindau tumor suppressor gene located on chromosome 3p25. Tumor development in this setting is due to inactivation or loss of the remaining wild-type allele in a susceptible cell. The highly vascular nature of VHL-associated neoplasms can be understood in light of the recent finding that the VHL gene product (pVHL) inhibits the accumulation of hypoxia-inducible mRNAs, such as the mRNA encoding vascular endothelial growth factor (VEGF), under normoxic conditions. This property of pVHL appears to be linked to its ability to bind to complexes containing elongin B, elongin C, and cullin 2 (Cul2). Elongin C and Cul2, based on their homology with Skp1 and Cdc53, respectively, are suspected of targeting certain proteins for covalent modification with ubiquitin and hence for degradation. One model, which remains to be tested, is that the binding of pVHL to elongins B/C and Cul2 affects the ubiquitination of RNA-binding proteins that regulate the stability of hypoxia inducible mRNAs. PMID- 9413425 TI - Fever of unknown origin (FUO). I A. prospective multicenter study of 167 patients with FUO, using fixed epidemiologic entry criteria. The Netherlands FUO Study Group. AB - Internal medicine wards in all 8 university hospitals in the Netherlands participated in this prospective study of fever of unknown origin (FUO) from January 1992 until January 1994 in order to update information on the spectrum of diseases causing FUO. We used fixed epidemiologic entry criteria to achieve completeness of enrollment and to avoid unintended selection bias. After entry, immunocompetent patients were included using criteria for FUO according to Petersdorf and Beeson (30). A standardized diagnostic protocol was used, and potentially diagnostic clues (PDCs) and their use in the diagnostic process were prospectively registered. Thus, the criteria of classic FUO have been adjusted to modern times: immunocompromised patients are excluded, and the time-criterion "1 week in hospital without a diagnosis" has been replaced by a quality-criterion stating that certain investigations must be performed as a minimum, and PDCs must be followed adequately for at least 1 week, without a diagnosis being reached. A total of 167 immunocompetent patients with FUO were thus retrieved, of whom 43 (25.7%) had infections, 21 (12.6%) had neoplasms, and 40 (24.0%) had noninfectious inflammatory diseases. No diagnosis was made in 50 patients (29.9%), 37 of whom recovered spontaneously. This study confirms the changing spectrum of diseases causing FUO. Indeed, as shown by another recent study, the group of patients with FUO in whom no diagnosis can be made is expanding, and mostly it concerns self-limiting or benign fevers. Others have suggested that this trend is not really occurring (29). We did not place patients with diseases of unknown origin in the "nondiagnosis" group, and indeed made presumptive diagnoses when necessary. Nevertheless, this category of undiagnosed fevers is increasing. We believe that the higher percentage of undiagnosed cases can be attributed to the greater use of advanced diagnostic techniques attendant on an increased number of self-limited illnesses in patients meeting criteria for FUO. Because of ongoing development in diagnostic techniques and the prospective influence on the spectrum of diseases causing FUO, studies should be performed regularly to update information on this subject. Because the number of outpatient evaluations for FUO is expected to increase, patients seen on an outpatient basis should be included in future studies. To avoid unwanted selection bias, fixed epidemiologic entry criteria should be used to ensure completeness of enrollment. To shorten the period of collecting data, multicentric studies can be done using standardized diagnostic protocols. In patients with recurrent fever or fever lasting longer than 6 months, the chance of reaching a diagnosis is significantly lower, and especially in this group one should exercise the greatest caution to avoid abundant and extensive diagnostic procedures. The diagnostic process in patients with FUO remains an intriguing problem in medicine. Recent microbiologic techniques may be useful as an approach to the relatively large proportion of patients in whom we now fail to make a diagnosis. PMID- 9413426 TI - Fever of unknown origin (FUO). II. Diagnostic procedures in a prospective multicenter study of 167 patients. The Netherlands FUO Study Group. AB - From January 1992 until January 1994, we used a standardized diagnostic protocol for the 167 immunocompetent patients with fever of unknown origin (FUO) admitted on the internal medicine wards in all 8 university hospitals in the Netherlands. This protocol consisted of a standardized coded history and standardized physical examination for all 167 patients. A number of additional obligatory investigations had to be performed in the first week of admission for all patients, and all potentially diagnostic clues (PDCs) thus retrieved had to be registered. In the presence of PDCs, specific investigations had to be performed based on the differential diagnosis. In the absence of PDCs or in the presence of only misleading PDCs, patients underwent a screening 2-staged diagnostic protocol. In 162 (97%) patients, PDCs were present after 1 week of admission. In 61 patients these PDCs were all misleading. The likelihood of reaching a diagnosis in patients with PDCs was not significantly higher than that in patients without PDCs, probably because of the high proportion of misleading PDCs. The likelihood of establishing a diagnosis was significantly lower (< 10%) only for patients with recurrent fever, normal erythrocyte sedimentation rate (ESR), and normal hemoglobin. All other PDCs were not significantly different in patients with a diagnosis compared with patients without a diagnosis. The screening 2-staged diagnostic protocol proved useful in 10 of 43 patients in whom it was used. The screening value of immunologic and microbiologic serology and endocrine investigations was nil; these investigations probably should be performed only when PDCs for the disease searched for are present. Scintigraphic techniques, echocardiography, and other imaging procedures were never helpful in our population in the absence of PDCs. Many patients with FUO had nonspecific anemia and disturbed liver chemistry. In the presence of these findings alone, without other more specific PDCs, the likelihood of reaching a diagnosis with help of bone marrow aspiration was nil, and with help of liver biopsy, it was low. Enteric biopsy was never helpful. If lymphadenopathy was confined to the cervical or inguinal region (with negative chest X-ray and abdominal ultrasound), lymph node biopsy was not helpful, in contrast to patients having generalized lymphadenopathy, in whom the technique had a yield of 79%. As shown in this study, the search for PDCs remains an important tool for establishing the diagnosis in patients with FUO, although in many cases these PDCs appear to be misleading. Directed diagnostic workup--using the PDCs retrieved by repeated, meticulous history taking and physical examination--remains the most efficient and intellectually satisfactory way to solve the problem of FUO in the individual patient. A standard protocol in patients with FUO in whom the obligatory investigations, as used by us, do not lead to the diagnosis can be limited to the tests that proved to be of some use as screening procedure: temporal biopsy in patients older than 55 years; fundoscopy; serology (Western blot) for Yersinia enterocolitica; serum for cryoglobulin at an early stage of the diagnostic process; and bone biopsy, liver biopsy, abdominal computed tomography (CT), and chest CT at a later stage. Repeating a thorough history-taking, physical examination, and obligatory investigations and waiting for PDCs to appear probably is better than ordering more screening investigations in the hope that something abnormal will come up. Supportive treatment with nonsteroidal anti inflammatory drugs (NSAIDs) can be helpful at this stage. Only rarely do patients deteriorate while using NSAIDs without presenting new PDCs. In these rare patients, further diagnostic workup should be performed or a therapeutic trial with, for example, antibiotics, steroids, or antituberculous agents started. PMID- 9413427 TI - Pneumocystis carinii infection in heart transplant recipients. Efficacy of a weekend prophylaxis schedule. AB - Most series of heart transplant patients report incidences of Pneumocystis carinii pneumonia (PCP) below 5% but do not individually describe the cases. From August 1988 to March 1994, 138 patients received 1 or more heart transplants at our institution. No anti-PCP chemoprophylaxis was provided, and 5 (3.6%) patients developed PCP. Incidence for listeriosis was 0.7% and for nocardiosis, 3.6%. We found descriptions of 14 more heart transplant patients with PCP in the medical literature. Data from the 19 patients follow. Mean age was 52 years, and PCP was diagnosed a median of 75 days after heart transplant (range, 37-781 d). Clinical presentation was acute (less than 48 h) with fever (89%), shortness of breath (84%), dry cough (74%), and hypoxia (63%). Cytomegalovirus was isolated from lung or blood in 74% of patients. Chest X-ray usually showed interstitial pneumonia (84%). Three patients required ventilatory support. All patients were treated with trimethoprim-sulfamethoxazole (TMP/SMX) (4 also with corticosteroids and 5 with ganciclovir). Mortality was 26%. Older age was the only significant poor prognostic factor (61 versus 49 years; p < 0.03). From March 1994, 50 heart transplant patients were given TMP/SMX prophylaxis at our institution (1 double strength tablet, 160/800 mg, every 12 hours on Saturdays and Sundays), and no new cases of PCP, Listeria or Nocardia have been detected since then. Tolerance has been excellent. Heart transplant recipients are at a substantial risk of PCP pneumonia, which presents with an abrupt onset and a high mortality. Weekend TMP/SMX chemoprophylaxis was very effective at our institution. PMID- 9413428 TI - Cerebral tuberculosis in patients with the acquired immunodeficiency syndrome (AIDS). Report of 6 cases and review. AB - Cerebral tuberculosis (TB) was diagnosed in 6 (4%) of 156 HIV-infected patients with TB seen at our institution over 6 years. We describe here the clinical and radiologic features of these cases and of 15 others reported in the literature. Of the 21 patients, 59% were intravenous drug users. Presenting symptoms were fever (76%), confusion (52%), seizures (38%), and headache (38%). Fourteen patients (66%) had previous or active extracerebral TB at presentation. Cranial CT scan showed ring-(62%) or nodular-(24%) enhancing lesions or mixed forms (14%). Among the 12 patients who underwent a brain biopsy, bacteriologic evidence of TB was found in 9. Four patients (19%) died during hospitalization. Among the 17 others who received antituberculous therapy, only 1 developed neurologic sequelae. Five patients also received steroid therapy to control cerebral edema or paradoxical growth of the cerebral mass lesions. TB should be considered as a cause of cerebral mass lesions in HIV-infected patients, especially if tuberculous infection is suspected at other sites. PMID- 9413429 TI - Cytotoxic therapy in systemic lupus erythematosus. Experience from a single center. AB - The present survey of cytotoxic therapy from a single large lupus clinic has shown that approximately 33% of the patients have received cytotoxic therapy at some point in their course. These agents were initiated for a variety of manifestations, with renal manifestations being the major indication, accounting for 28.2% of the cytotoxic agents used. Other common indications for initiation of cytotoxic therapy included steroid sparing (18.4%), global flare (12.5%), neurologic manifestations (11.4%), and musculoskeletal (8.6%). Azathioprine, methotrexate, and cyclophosphamide accounted for 98% of all cytotoxic agents used. Azathioprine was the most frequently used cytotoxic drug (70%), followed by methotrexate (21.5%) and cyclophosphamide (9.4%). Cytotoxic agents were used sequentially in 12.5% of patients and in combination in 4.2% of the patients. Overall, the use of cytotoxic therapy appears to be beneficial in reducing global disease activity, as the mean SLEDAI fell by 2.59 (33%) over 6 months of cytotoxic therapy, and the mean steroid dose was reduced by 37% over the same time period. There was also an improvement in most organ-specific indications with the use of cytotoxic agents. Overall the cytotoxic agents were well tolerated, with 17% of the courses being discontinued due to a side effect. Cytopenia was the most common side effect necessitating discontinuation of cytotoxic agents. PMID- 9413430 TI - A defect in GTP synthesis affects mannose outer chain elongation in Saccharomyces cerevisiae. AB - We have found that yeast mutants that are defective in mannose outer chain elongation of N-linked glycoproteins show higher cell wall porosity than normal cells, and are hypersensitive to antibiotics with a large molecular weight; such as neomycin and geneticin. Wild-type yeast cells also showed enhanced sensitivity to neomycin in the presence of tunicamycin, an inhibitor of N-glycosylation, suggesting that the extent of N-glycosylation may affect the sensitivity of yeast cells to drugs and that sensitivity to neomycin may be an effective method for screening for yeast mutants defective in N-glycosylation. Pursuing this logic, we isolated neomycin-sensitive yeast mutants and screened them for defects in N glycosylation. The neomycin-sensitive, N-glycosylation-defective mutants fell into 15 complementation groups including alleles of the previously isolated temperature-sensitive nes mutants nes10, nes17, and nes25. Gene cloning revealed that NES10 was identical to SEC20, which is involved in ER-Golgi protein transport, NES17 was identical to ALG1, which encodes a beta-1,4 mannosyltransferase present in the ER, MSN17, a multicopy suppressor of nes17/alg1, was also isolated and found to be an allele of PSA1, which is involved in GDP-mannose synthesis, NES25 was identical to GUK1, which encodes a GMP kinase. Overexpression of MSN17 increased the GDP-mannose level in a wild type strain by about threefold, and guk1 decreased the GDP-mannose level to one fourth, suggesting a close relationship between GTP metabolism and mannose outer chain elongation; the link is presumably provided by the process of GDP-mannose transport in the Golgi membranes. PMID- 9413431 TI - Yeast Crv4/Ttp1, a predicted type II membrane protein, is involved in an event important for growth, functionally overlapping with the event regulated by calcineurin- and Mpk1-mediated pathways. AB - The Saccharomyces cerevisiae crv mutants (crv1, 2, 3 and 4) exhibit phenotypes, such as calcium resistance and vanadate sensitivity, which are apparently similar to those of calcineurin-deficient mutants. We have cloned and characterized the CRV4 gene that complements all the phenotypes of the crv4 mutant. DNA sequencing revealed that CRV4 is identical to the previously cloned gene TTP1, which encodes a type II membrane protein of unknown function. Deletion of CRV4/TTP1 causes no obvious phenotype except for Ca2+ resistance and vanadate sensitivity, but is synthetically lethal in combination with a deletion of MPK1, in a manner which is suppressible by the addition of an osmotic stabilizer. In medium containing sorbitol as an osmotic stabilizer, the enb1 mpk1 ttp1 triple mutant exhibits a more severe growth defect than does any of the double mutants enb1 ttp1, enb1 mpk1 or mpk1 ttp1. A high Ca2+ concentration (50 mM) or a constitutively active form of calcineurin partially suppresses the growth defect of the mpk1 ttp1 double mutant. These results indicate that Ttp1 participates in a cellular event essential for growth and morphogenesis, in parallel with the pathways involving Mpk1 MAP kinase and calcineurin. PMID- 9413432 TI - Molecular and functional analysis of the ribosomal L11 and S12 protein genes (rplK and rpsL) of Streptomyces coelicolor A3(2). AB - A RelC deletion mutant, KO-100, of Streptomyces coelicolor A3(2) has been isolated from a collection of spontaneous thiostrepton-resistant mutants. KO-100 grows as vigorously as the parent strain and possesses a 6-bp deletion within the rplK, previously termed relC. When the wild-type rplK gene was propagated on a low-copy-number vector in mutant KO-100, the ability to produce ppGpp, actinorhodin and undecylprodigiosin, which had been lost in the RelC mutant, was completely restored. Allele replacement by gene homogenotization demonstrated that the RelC mutation is responsible for the resistance to thiostrepton and the inactivation of ppGpp, actinorhodin and undecylprodigiosin production. Western blotting showed that ribosomes from the RelC mutant KO-100 contain only one eighth the amount of L11 protein found in ribosomes of the parent strain. The impairment of antibiotic production in KO-100 could be rescued by the introduction of mutations that confer resistance to streptomycin (str), which result in alteration of Lys-88 in ribosomal protein S12 to Glu or Arg. No accompanying restoration of ppGpp synthesis was detected in these RelC str double mutants. PMID- 9413433 TI - Genetic stability of gene-targeted immunoglobulin loci. II. Influence of the cell line and the vector linearization site. AB - The site-specific integration of exogenous gene fragments by homologous recombination provides a convenient method for altering the immunoglobulin loci of B cells and specifically designing antibody molecules. To introduce a human isotype into the heavy chain locus of mouse hybridoma cells we compared the recombination frequencies of vectors that could be linearized either as integration or as replacement constructs in different cell lines. Integration as well as replacement recombination was observed, irrespective of the location of the site at which the vector was cleaved. Integration events involving the human IgG1 vectors were lost at high frequency due to secondary vector excision, so that all stable recombinations were found to be replacement events. Replacement recombination of an integration vector involves an illegitimate crossover at least at the 3' side and sometimes gives rise to deletion of the CH1 domain. However, a homologous event at the 3' side is more efficient than an illegitimate one, so that a homology that is distributed on both sides of the heterologous region promotes targeting at higher frequency than a contiguous sequence of the same total length. The position of the linearization site in the vector markedly influenced the targeting efficiency, but surprisingly, whether a double-strand break in the homology or in the heterology region more efficiently promoted integration was dependent on the cell line. In all cells, however, cleavage of the vector outside the homology region favoured stable replacements with a bias against CH1-truncated clones. We further show that the frequency of replacements induced by integration vectors is not correlated to the homology length and cannot be increased by irradiation of the cells. Our findings indicate that for targeting the IgH locus other mechanisms might be involved than at other loci. PMID- 9413434 TI - Physical mapping of RFLP markers on four chromosome arms in maize using terminal deficiencies. AB - Terminal deficiencies (TDs) generated by the r-X1 deletion system in maize were used to physically map RFLP markers on the short arm of chromosome 2 (2S) and the long arm of chromosome 6 (6L), chromosome 8 (8L), and chromosome 10 (10L). Five TDs on 2S, 8 on 6L, 10 on 8L, and 20 on 10L were isolated using the recessive morphological markers lg1, py1, j1(gl18), and sr2, respectively, for selection. Two exceptional TDs on 2S and 8L also have a second breakpoint on the long arm of chromosome 2 (2L) and 8L, respectively. The physical mapping of RFLP probes in relation to TD breakpoints was done by Southern hybridization. The five TDs on 2S divide chromosome 2 into four regions, all of which are distinguishable by RFLP markers. Likewise, three remaining chromosome arms are divided by TDs into RFLP marked regions: 8 TDs divide 6L into five regions, 10 TDs divided 8L into seven regions, and 20 TDs divide 10L into three regions. The linear order of the physical map of 6L and 8L is consistent with that of the genetic maps, but that of 2L and 10L is not. Four groups of markers on 2S as well as 2L, and two on 10L are in reverse order in the physical map compared with the genetic maps. Other intriguing results are that breakpoints of TDs on 6L and 8L are distributed throughout the selected region, but most of those on 2L and 10L cluster in a region near the centromere: a single TD arose after fertilization. PMID- 9413436 TI - The Neisseria gonorrhoeae gene aniA encodes an inducible nitrite reductase. AB - The aniA gene of Neisseria gonorrhoeae encodes an outer membrane lipoprotein which is strongly induced when gonococci are grown anaerobically in vitro in the presence of nitrite. Database searches with the amino acid sequence derived from the aniA structural gene revealed significant homologies to copper-containing nitrite reductases from several denitrifying bacteria. We constructed an insertional mutation in the aniA locus of strain MS11 by allelic replacement, to determine whether this locus was necessary for growth in oxygen-depleted environments, and to demonstrate that AniA was indeed a nitrite reductase. The mutant was severely impaired in its ability to grow micro-aerophilically in the presence of nitrite, and we observed a loss in viability over several hours of incubation. No measurable nitrite reductase activity was detected in the aniA mutant strain, and activity in the strain with a wild-type locus was inducible. Finally, we report investigations to determine whether AniA protein is involved in gonococcal pathogenesis. PMID- 9413435 TI - Southern analysis of BT-R1, the Manduca sexta gene encoding the receptor for the Cry1Ab toxin of Bacillus thuringiensis. AB - Various subspecies of the gram-positive bacterium Bacillus thuringiensis are known to produce a wide array of insecticidal crystal proteins (ICPs) upon sporulation. These ICPs act primarily on the brush border of midgut epithelial cells of susceptible larvae. Recently, a protein of 210 kDa, isolated from the midgut of Manduca sexta, has been demonstrated to bind the Cry1Ab toxin produced by B. thuringiensis subsp, berliner and is therefore postulated to be involved in mediating the toxicity of Cry1Ab. The cDNA encoding the 210 kDa protein, termed BT-R1 (Bacillus thuringiensis receptor-1), was recently cloned, and shows limited homology to the cadherin superfamily of proteins. Quite naturally, there is a great deal of interest in the characterization of BT-R1, the gene encoding the 210 kDa Cry1Ab binding protein. The studies presented here involve the use of various restriction fragments prepared from the cDNA encoding BT-R1 as probes of Southern blots bearing M. sexta genomic DNA cleaved with a variety of restriction endonucleases. These Southern blot data reveal that there are two discrete regions within the M. sexta genome which encode sequences homologous to BT-R1. On the basis of the signal intensities seen on Southern blots, it appears that only one of these genes encodes BT-R1, whereas the other is a closely related homologue. PMID- 9413437 TI - The processing of DNA ends at double-strand breaks during homologous recombination: different roles for the two ends. AB - We have investigated the role of DNA ends during gap repair by homologous recombination. Mouse cells were transfected with a gapped plasmid carrying distinctive ends: on one side mouse LINE-1 repetitive sequences (L1Md-A2), and on the other rat LINE-1 sequences (L1Rn-3). The gap could be repaired by homologous recombination with endogenous mouse genomic LINE-1 elements, which are on average 95% and 85% homologous to L1Md-A2 and L1Rn-3 ends, respectively. Both L1Md-A2 and L1Rn-3 ends were found to initiate gap repair with equal efficiency. However, there were two types of gap repair products--precise and imprecise--the occurrence of which appears to depend on which end had been used for initiation and thus which end was left available for subsequent steps in recombination. These results, together with sequence analysis of recombinants obtained with plasmids having either mouse or rat LINE-1 sequences flanking the gap, strongly suggest that the two DNA ends played different roles in recombinational gap repair. One end was used to initiate the gap repair process, while the other end was involved at later steps, in the resolution of the recombination event. PMID- 9413438 TI - Structure and function of the vp1 gene homologue from the resurrection plant Craterostigma plantagineum Hochst. AB - A gene that is homologous to the vp1 genes from maize and rice, the Arabidopsis abi3 gene and PvAlf from Phaseolus vulgaris was isolated from the resurrection plant, Craterostigma plantagineum Hochst, The C. plantagineum gene (epvp1) encodes a protein of 688 amino acids and contains five introns at positions identical to those in the Arabidopsis, maize and rice homologues. The cpvp1 transcript is present in mature seeds and in young seedlings up to 8 days following germination. The ability of the cpvp1 gene product to activate target genes was demonstrated by transient expression experiments in tobacco protoplasts using reporter gene constructs containing the beta-glucuronidase (GUS) gene fused to the promoter of two late embryogenesis abundant (LEA)-like genes, CDeT27-45 and CDeT6-19 from C. plantagineum, or the promoter of a maize gene encoding a 22 kDa zein seed storage protein. PMID- 9413439 TI - The yeast FET5 gene encodes a FET3-related multicopper oxidase implicated in iron transport. AB - The yeast FET3 gene encodes an integral membrane multicopper oxidase required for high-affinity iron uptake. The FET4 gene encodes an Fe(II) transporter required for low-affinity uptake. To identify other yeast genes involved in iron uptake, we isolated genes that could, when overexpressed, suppress the iron-limited growth defect of a fet3 fet4 mutant. The FET5 gene was isolated in this screen and it encodes a multi-copper oxidase closely related to Fet3p. Several observations indicate that Fet5p plays a role analogous to Fet3p in iron transport. Suppression of the fet3 fet4 mutant phenotype by FET5 overexpression required the putative FTR1 transporter subunit of the high-affinity system. Fet5p is an integral membrane protein whose oxidase domain is located on the cell surface or within an intracellular compartment. Oxidase activity measured in cells with altered levels of FET5 expression suggested that Fet5p is a functional oxidase. FET5 overexpression increased the rate of iron uptake by a novel uptake system. Finally, FET5 mRNA levels are regulated by iron and are increased in cells grown in iron-limited media. These results suggest that Fet5p normally plays a role in the transport of iron. PMID- 9413440 TI - Controls of the expression of aspA, the aspartyl protease gene from Penicillium roqueforti. AB - The gene (aspA) encoding the extracellular aspartyl protease from Penicillium roqueforti was cloned and characterized. Northern hybridization analyses and beta casein degradation assays revealed that aspA was strongly induced by casein in the medium and efficiently repressed by ammonia. External alkaline pH overrides casein induction, resulting in aspA repression. Cis-acting motifs known to mediate nitrogen and pH regulation of fungal gene expression are present in the aspA promoter and protein-DNA binding experiments showed that mycelial proteins interact with various regions of the promoter. Due to the efficient environmental controls on aspA expression, the promoter of aspA is an attractive candidate for the development of a controllable gene expression system in P. roqueforti. PMID- 9413441 TI - Movement of a small mitochondrial double-stranded RNA element of Cryphonectria parasitica: ascospore inheritance and implications for mitochondrial recombination. AB - Movement via somatic fusion and inheritance of a small mitochondrial double stranded (ds) RNA element was examined in Cryphonectria parasitica. The 2.7-kb dsRNA from the C. parasitica strain NB631 encodes a putative RNA-dependent RNA polymerase when the mitochondrial code (UGA = Trp) is invoked. All progeny from asexual spores (conidia) of strain NB631 examined for dsRNA contained the 2.7-kb element. Unlike other C. parasitica dsRNAs, which are cytoplasmic, the dsRNA in strain NB631 was transmitted through the sexual cycle (ascospores) if the strain containing the element acted as the female in crosses. Movement of the 2.7-kb dsRNA was also observed through hyphal anastomosis. Transfer by anastomosis was accompanied by mitochondrial movement and recombination of the mitochondrial genome as determined by RFLP analysis. In control pairings between isolates lacking dsRNA, mitochondrial movement and recombination were also observed. Transfer by anastomosis allowed the generation of infected and uninfected isogenic lines, and permitted us to evaluate the effects of the dsRNA element on virulence of the host. Bark virulence assays on American chestnut suggest that NB631 dsRNA decreases the virulence of C. parasitica, but not to the level associated with members of the Hypoviridae. PMID- 9413442 TI - Cell wall biogenesis in Chlamydomonas: molecular characterization of a novel protein whose expression is up-regulated during matrix formation. AB - In the unicellular eukaryote Chlamydomonas, disruption of cell-matrix interactions by treatment with a periplasmic matrix metalloproteinase, g-lysin, activates a signal transduction pathway that results in the rapid synthesis and secretion of matrix molecules, followed by their assembly into a new matrix. I have identified and partially characterized several cDNA clones for transcripts that are dramatically up-regulated following treatment of cells with g-lysin. Here I report the complete nucleotide sequence and preliminary characterization of a matrix-related molecule termed Mrp47. The cDNA clone for Mrp47 contained an insert of 2.5 kb, corresponding to a transcript of 3.0 kb that is encoded by a single-copy gene. Sequence analysis indicated that Mrp47 cDNA contains an open reading frame (ORF) that encodes a 46-kDa polypeptide. The putative polypeptide is unusually rich in the amino acids proline, alanine and serine, with prolines clustered together in a 30-amino acid N-terminal region and a 80-amino acid C terminal region. Further analysis of the predicted amino acid sequence suggested that Mrp47 is likely to be a secreted glycoprotein. Southern hybridization analysis indicated that Mrp47 is encoded by a single-copy gene in the Chlamydomonas genome. Database searches suggested that Mrp47 shows homology to other proline-rich proteins including a surface glycoprotein in Volvox and verprolin from yeast. PMID- 9413443 TI - Silencing of transgenes introduced into leaves by agroinfiltration: a simple, rapid method for investigating sequence requirements for gene silencing. AB - Agroinfiltration--the infiltration of Agrobacterium tumefaciens into intact plant levels--provides a rapid and simple way of screening large numbers of transgene constructs for silencing in response to a resident transgene. Transgenic Nicotiana sylvestris plants homozygous for the tobacco class I chitinase A gene CHN48 under the control of the cauliflower mosaic virus 35S RNA promoter (P35S) show a high incidence of postranscriptional gene silencing. We forced suspensions of A. tumefaciens, carrying P35S-CHN48 in a binary Tiplasmid vector, into wild type and transgenic N, sylvestris leaves with a blunt-tipped plastic syringe. The infiltrated CHN48 transgene was expressed in leaves transformed with the vector alone, but not in CHN48-transformed leaves showing the silent phenotype. In contrast, expression of a chimeric P35S-E. coli beta-glucuronidase gene (uidA) infiltrated into leaves was not affected by the presence of the CHN48 transgene stably integrated in the host genome. These results show that extra copies of CHN48 are silenced by resident, silent copies of the same gene and confirm that CHN48 silencing is not the result of promoter interactions. The results also suggest that silencing of the additional CHN48 copies does not require their integration into chromosomes. PMID- 9413444 TI - Cloning with Mud-P22 hybrid prophages: mapping of IS200 elements on the chromosome of Salmonella typhimurium LT2. AB - Specific DNA fragments from the chromosome of Salmonella typhimurium LT2 were packaged in P22 capsids by induction of "locked-in" Mud-P22 hybrid prophages. High yields of the packaged DNA were obtained upon capsid disruption. DNA hybridization using a fragment of insertion sequence IS200 as probe permitted physical mapping of IS200 elements on the chromosome of S. typhimurium LT2 within +/- 1 centisome (CS). IS200 copies were found at the following locations: CS 24 (copy VI), CS 53 (copy V), CS 63 (copy I), CS 80 (copy II) and CS 93 (copy III). Copy IV, previously mapped near fliA (CS 42), was not included in our study. PMID- 9413445 TI - Interorganellar gene transfer in bryophytes: the functional nad7 gene is nuclear encoded in Marchantia polymorpha. AB - The nad7 gene, encoding subunit 7 of NADH dehydrogenase, is mitochondrially encoded in seed plants. In the liverwort, Marchantia polymorpha, only a pseudogene is located in the mitochondrial genome. We have now identified the functional nad7 gene copy in the nuclear genome of Marchantia, coding for a polypeptide of 468 amino acids. The nuclear-encoded nad7 has lost the two group II introns present in the mitochondrial pseudogene copy. Instead, a typical nuclear intron is found to split an exon encoding the presumptive mitochondrial targeting signal peptide and the mature subunit 7 of NADH dehydrogenase. These results suggest that RNA-mediated gene transfer from the mitochondrial into the nuclear genome occurs not only in seed plants but also in bryophytes. PMID- 9413446 TI - "Break on through". PMID- 9413447 TI - Effects of positioning and exercise on intracranial pressure in a neurosurgical intensive care unit. AB - BACKGROUND AND PURPOSE: The purpose of the study was to assess the safety of physical therapy by investigating its effects on intracranial pressure (ICP) and cerebral perfusion pressure. SUBJECTS: The subjects were 65 patients in a neurosurgical intensive care unit who had normal ICP (< 15 mm Hg) or increased ICP (> 15 mm Hg). METHODS: Intraventricular ICP was measured in a 30-degree head up position (all patients) and in supine and 45-degree head-up positions (patients with normal ICP) during passive range of motion (comatose patients) and exercises involving limb movement (awake patients). RESULTS: In patients with normal ICP, passive range of motion decreased mean ICP by 1 mm Hg in the supine position but not in the head-up position. In patients with high ICP, it decreased ICP by 2 mm Hg. Limb exercises left the mean ICP essentially unchanged in both the patients with normal ICP and the patients with high ICP. Isometric hip adduction increased mean ICP by 4 mm Hg in patients with normal ICP. It did not affect ICP in patients with high ICP. Limb movement was associated with suppression of abnormal ICP waves and improvement of consciousness in 13 patients. CONCLUSION AND DISCUSSION: Physical therapy can be used safely in patients with normal or increased ICP provided that Valsalva-like maneuvers are avoided. [Brimioulle S, Moraine J-J, Norrenberg D, Kahn RJ. Effects of positioning and exercise on intracranial pressure in a neurosurgical intensive care unit. PMID- 9413448 TI - A comprehensive treatment approach for patellofemoral pain syndrome in young women. AB - BACKGROUND AND PURPOSE: The purposes of this study were (1) to evaluate a comprehensive treatment approach for patients with patellofemoral pain syndrome and (2) to compare a training program using isometric muscle contractions with a training program using eccentric muscle contractions. SUBJECTS: Forty female patients with patellofemoral pain syndrome, aged 15 to 28 years (mean = 20.2, SD = 3.2), were randomly assigned to either a group using isometric muscle contractions or a group using eccentric muscle contractions. METHODS: The effects of 12 weeks of treatment, consisting of an educational component and a training program, on physical activity, pain, and muscle function were evaluated after 3 and 12 months. RESULTS: No differences were found between the two groups, except in one of the torque measurements. A reduction in pain and improvements in torque, vertical jumping ability, and physical activity level were seen in both groups after treatment. At the 12-month follow-up, 85% of the subjects were participating in sports without pain and 37 subjects rated their overall knee function as excellent or good. CONCLUSION AND DISCUSSION: The results indicate that the improvements shown in this study may be due to spontaneous recovery over time, the education given to the subjects, the pain monitoring system, the gradually progressing training program, and the adjusted physical activity. [Thomee R. A. Comprehensive treatment approach for patellofemoral pain syndrome in young women. PMID- 9413449 TI - Health-related quality of life for patients with progressive multiple sclerosis: influence of rehabilitation. AB - BACKGROUND AND PURPOSE: The health-related quality of life of patients with multiple sclerosis (MS) is an important aspect of care outcome. The purpose of this study was to compare health-related quality of life between patients who received weekly comprehensive outpatient rehabilitation for 1 year and a group that did not receive rehabilitation. SUBJECTS: Twelve patients receiving outpatient care for chronic progressive MS (mean age = 44.5 years, SD = 11.6) were compared with 19 similar patients (mean age = 49.2 years, SD = 9.2) on a waiting list who were not receiving outpatient care. METHODS: A pretest-posttest longitudinal design was used to descriptively compare outcome measures. Multivariate regression analyses were used to determine which variables, controlled for baseline health status and other relevant patient characteristics, were related to the best outcomes at the time of the 1-year follow-up. RESULTS: The treatment group showed improvements in six health status measures on the Rand 36-Item Health Survey 1.0 (SF-36) that were not improved in the wait-listed group. Outpatient treatment was the sole predictor of positive outcome for energy/fatigue (partial R2 = .43) and change in general health (partial R2 = .19). In addition, the treatment group was associated with a positive outcome (together with other independent variables) in the domains of social function and social support. CONCLUSION AND DISCUSSION: Patients with chronic progressive forms of MS appear to derive benefits from an ongoing comprehensive outpatient rehabilitation program. [Di Fabio RP, Choi T, Soderberg J, Hansen CR. Health related quality of life for patients with progressive multiple sclerosis: influence of rehabilitation. PMID- 9413450 TI - Estimates of weight that subjects can lift frequently in functional capacity evaluations. AB - BACKGROUND AND PURPOSE: This study was designed to determine whether the maximum weight that can be lifted during 33% to 67% of a workday (weight lifted frequently) can be accurately predicted from other data gathered during a functional capacity evaluation (FCE). The best equations for estimating weight lifted frequently were also identified, and the interrater reliability for measurements obtained during a 6-day FCE was calculated. SUBJECTS: A retrospective chart audit was conducted of clients (36 female, 93 male) who completed 22-hour FCEs during a 6-day period. Ninety subjects had spinal problems, and 39 subjects had injuries not involving the spine. Subjects were randomly assigned to a model estimation group (n = 109) or a cross-validation group (n = 20). METHOD: A stepwise multiple regression analysis identified the best predictors of weight lifted frequently, with subjects lifting from four different heights. The resulting equations were used to predict the weight for the cross-validation group, and predicted weights were compared with observed weights using t tests and correlation coefficients. RESULTS: Regression equations explained between 27.3% and 93.0% of the variance in weight, with standard errors of estimation between 1.09 and 5.72. No differences between the mean observed and predicted scores were found when estimates were based on weight only occasionally lifted (maximum weight that could be lifted up to 33% of a workday). CONCLUSION AND DISCUSSION: Estimates of weight lifted frequently based on weight lifted occasionally can be made, but the usefulness of these estimates is questionable. Clinicians should use caution when using estimations of weight lifted for return to-work recommendations. [Saunders RL, Beissner KL, McManis BG. Estimates of weight that subjects can lift frequently in functional capacity evaluations. PMID- 9413451 TI - Effect of high-voltage pulsed current and alternating current on macromolecular leakage in hamster cheek pouch microcirculation. AB - BACKGROUND AND PURPOSE: Electrical stimulation (ES) is supposed to affect edema formation by inhibiting macromolecular leakage from microvessels. The purpose of the study was to determine the effects of various forms of ES on macromolecular leakage from microvessels. SUBJECTS: Fifty-three hamsters were randomly assigned to one of seven groups: a control group (histamine only); groups that received histamine with cathodal high-voltage pulsed current (HVPC) at intensities of 90%, 50%, and 10% of visible motor threshold (VMT); groups that received anodal HVPC at intensities of 90% and 50% of VMT; and a group that received alternating current (AC) at 90% of VMT. METHODS: Anesthetized animals were injected with fluorescein-labeled dextran. Macromolecular leakage was determined by computer analysis of fluorescence microscopy images for 5 minutes after treatment. RESULTS: When compared with controls, leakage was less in groups treated with cathodal HVPC at 90% and 50% of VMT and anodal HVPC at 90% of VMT. CONCLUSION AND DISCUSSION: Cathodal and anodal HVPC, but not AC, curb macromolecular leakage from the microvessels of histamine-treated hamsters. [Taylor K, Mendel FC, Fish DR, et al. Effect of high-voltage pulsed current and alternating current on macromolecular leakage in hamster cheek pouch microcirculation. PMID- 9413452 TI - Accuracy of reconstructed angular estimates obtained with the Ariel Performance Analysis System. AB - BACKGROUND AND PURPOSE: Three-dimensional computerized gait analysis continues to grow in use among physical therapists and other clinical specialists interested in quantitative data regarding human ambulation. This study documented the accuracy of reconstructed angular estimates under static and dynamic conditions using the Ariel Performance Analysis System. METHODS: Angular velocity was systematically increased by raising the release position of a T-shaped pendulum. Angular velocities were examined by releasing the pendulum from four angles (0 degree-static, 45 degrees, 90 degrees, and 120 degrees). Twelve reference angles were estimated over 20 autodigitized frames for 10 trials at each release position. Intraclass correlation coefficients (ICCs) and analysis-of-variance (ANOVA) procedures were used to test the hypothesis that the error of angular estimates grows with increasing angular velocity. RESULTS: Mean errors of the reconstructed angles were consistently within +/- 1.0 degree, regardless of angular velocity. An ANOVA revealed a statistically significant angular velocity effect, characterized by release position. The 90-degree release position produced the greatest error, followed by the 120-, 45-, and 0-degree release positions. The error was not significantly different between the 120- and 45 degree release positions. Intraclass correlation coefficients greater than .90 were found for all frame-to-frame angular velocities, except for the 90-degree release position. The angle estimates consistently underestimated the reference angles, regardless of release position. CONCLUSION AND DISCUSSION: The results suggest that clinically accurate angular estimates can be obtained across the range of angular velocities used in this study. PMID- 9413453 TI - Open versus closed kinetic chain exercise: issues in rehabilitation after anterior cruciate ligament reconstructive surgery. AB - What has been called "closed kinetic chain" (CKC) exercise has become popular in the last 5 to 10 years for use after anterior cruciate ligament (ACL) reconstructive surgery. Closed kinetic chain exercises appear to have gained popularity over more traditionally used open kinetic chain (OKC) exercises because many clinicians believe that CKC exercises are safer and more functional. These clinicians also contend that CKC exercise is equally effective as OKC exercise in restoring quadriceps femoris muscle force production following ACL reconstructive surgery. The purpose of this clinical perspective is to examine the evidence concerning OKC and CKC training after ACL reconstructive surgery with regard to these issues and discuss how physical therapists can best apply this knowledge in clinical practice. Based on the review of data, it does not appear that clinicians should completely abandon more traditional OKC exercises and replace them with CKC exercises in postoperative ACL reconstruction rehabilitation programs. Both types of exercise apparently can be modified to minimize (1) the risk of applying excessive strain on the ACL graft and (2) the risk of excessive patellofemoral joint stress. Depending on the functional goals of the patient, both OKC and CKC exercises may be appropriate for simulating functional activities. When improvement in quadriceps femoris muscle function is an essential treatment goal, therapists may need to combine OKC exercises with CKC exercises to provide optimal training stimuli. Suggestions for further research are discussed. [Fitzgerald GK. Open versus closed kinetic chain exercise: issues in rehabilitation after anterior cruciate ligament reconstructive surgery. PMID- 9413454 TI - Learning and understanding the Kruskal-Wallis one-way analysis-of-variance-by ranks test for differences among three or more independent groups. AB - When several treatment methods are available for the same problem, many clinicians are faced with the task of deciding which treatment to use. Many clinicians may have conducted informal "mini-experiments" on their own to determine which treatment is best suited for the problem. These results are usually not documented or reported in a formal manner because many clinicians feel that they are "statistically challenged." Another reason may be because clinicians do not feel they have controlled enough test conditions to warrant analysis. In this update, a statistic is described that does not involve complicated statistical assumptions, making it a simple and easy-to-use statistical method. This update examines the use of two statistics and does not deal with other issues that could affect clinical research such as issues affecting credibility. For readers who want a more in-depth examination of this topic, references have been provided. The Kruskal-Wallis one-way analysis-of variance-by-ranks test (or H test) is used to determine whether three or more independent groups are the same or different on some variable of interest when an ordinal level of data or an interval or ratio level of data is available. A hypothetical example will be presented to explain when and how to use this statistic, how to interpret results using the statistic, the advantages and disadvantages of the statistic, and what to look for in a written report. This hypothetical example will involve the use of ratio data to demonstrate how to choose between using the nonparametric H test and the more powerful parametric F test. PMID- 9413455 TI - Strength or endurance? PMID- 9413456 TI - Leprosy beyond the year 2000. PMID- 9413457 TI - Cytomegalovirus: the taming of the beast? PMID- 9413458 TI - Early indicators in acute dengue infection. PMID- 9413459 TI - Pyoderma gangrenosum and its treatment. PMID- 9413460 TI - Kangaroo mother care. PMID- 9413461 TI - An urban agenda for Europe. PMID- 9413462 TI - Risks of breast and testicular cancers in young adult twins in England and Wales: evidence on prenatal and genetic aetiology. AB - BACKGROUND: Aetiology of breast and testicular cancers may have prenatal factors, possibly exposure of the fetus to high concentrations of maternal oestrogen. Dizygotic twinning probably involves high hormone concentrations, and therefore, dizygotic twins might be at raised risk of these cancers. The aetiologies of breast and testicular cancers have genetic components, for breast cancer, especially at younger ages. Twins of these probands may, therefore, be at high risk. We investigated risk in twins of patients with breast cancer at young ages or with testicular cancer. METHODS: We identified twins with breast cancer incident at ages younger than 45 years and with incident testicular cancer in England and Wales during 1971-89 by cross-matching national cancer-registration and births records. We determined zygosity by questionnaires to the patients. The twins of probands were followed up for cancer incidence and death. We analysed risks of breast and testicular cancer in dizygotic twins compared with monozygotic twins, and in monozygotic and dizygotic twins of probands. FINDINGS: We identified 500 twins with breast cancer and 194 with testicular cancer. We found a non-significantly raised risk of breast cancer in dizygotic compared with monozygotic twins younger than 30 years (odds ratio 2.3 [95% CI 0.9-5.9]) but not older. The overall risk of testicular cancer was significantly higher in dizygotic twins than in monozygotic twins (1.5 [1.1-2.2]) consequent on a risk for seminomas was high (3.2 [1.6-6.5]; p = 0.001). Risk of breast cancer was significantly raised in female twins of probands (standardised incidence ratio 7.7 [4.9-12.2], p < 0.001). The relative risk of breast cancer was 34.7 (9.5 126.5) in monozygotic twins of women in whom breast cancer had occurred before age 35 years. The cumulative risk of breast cancer for these twins by age 40 years was 29% (13-56). The relative risk of testicular cancer was 37.5 (12.3 115.6) in twins of men with testicular cancer. The cumulative risk by age 40 years in monozygotic twins of men with testicular cancer was 14% (4-46). INTERPRETATION: The higher risks of these cancers in dizygotic than in monozygotic twins support a prenatal aetiology, and are compatible with aetiology related to raised maternal concentrations of free, unbound oestrogens. The results for twins of probands have implications for genetic aetiology; appropriate clinical action for monozygotic twins needs consideration. PMID- 9413463 TI - Randomised trial of efficacy and safety of oral ganciclovir in the prevention of cytomegalovirus disease in liver-transplant recipients. The Oral Ganciclovir International Transplantation Study Group [corrected]. AB - BACKGROUND: Cytomegalovirus (CMV) disease is a frequent cause of serious morbidity after solid-organ transplantation. The prophylactic regimens used to prevent CMV infection and disease have shown limited benefit in seronegative recipients. We studied the safety and efficacy of oral ganciclovir in the prevention of CMV disease following orthotopic liver transplantation. METHODS: Between December, 1993, and April, 1995, 304 liver-transplant recipients were randomised to receive oral ganciclovir 1000 mg or matching placebo three times a day. Seronegative recipients of seronegative livers were excluded. Study drug was administered as soon as the patient was able to take medication by mouth (no later than day 10) until the 98th day after transplantation. Patients were assessed at specified times throughout the first 6 months after surgery for evidence of CMV infection, CMV disease, rejection, opportunistic infections, and possible drug toxicity. FINDINGS: The Kaplan-Meier estimate of the 6-month incidence of CMV disease was 29 (18.9%) of 154 in the placebo group, compared with seven (4.8%) of 150 in the ganciclovir group (p < 0.001). In the high-risk group of seronegative recipients (R-) of seropositive livers (D+), incidence of CMV disease was 11 (44.0%) of 25 in the placebo group, three (14.8%) of 21 in the ganciclovir group (p = 0.02). Significant benefit was also observed in those receiving antibodies to lymphocytes, where the incidence of CMV disease was 12 (32.9%) of 37 in the placebo group and two (4.6%) of 44 in the ganciclovir group (p = 0.002). Oral ganciclovir reduced the incidence of CMV infection (placebo 79 [51.5%] of 154; ganciclovir 37 [24.5%] of 150; p < 0.001) and also reduced symptomatic herpes-simplex infections (Kaplan-Meier estimates: placebo 36 [23.5%] of 154; ganciclovir five [3.5%] of 150; p < 0.001). INTERPRETATION: Oral ganciclovir is a safe and effective method for the prevention of CMV disease after orthotopic liver transplantation. PMID- 9413464 TI - Sodium sensitivity and cardiovascular events in patients with essential hypertension. AB - BACKGROUND: In patients with sodium-sensitive hypertension, glomerular pressure is increased and microalbuminuria, a marker of glomerular hypertension, is a predictor of cardiovascular events. Similarly, the lack of a nocturnal decrease in blood pressure in these patients is also associated with an increased risk of cardiovascular events. We hypothesised that sodium sensitivity may be the common factor and carried out a retrospective study of cardiovascular events in patients with essential hypertension who had had sodium sensitivity measured in our clinic. METHODS: Sodium sensitivity was assessed in about 350 patients with essential hypertension during the initial investigation of their disorder. The definition of sodium sensitivity was a 10% or greater difference in blood pressure on low-sodium or high-sodium diets. By alphabetical order, the records of 201 patients were obtained and 156 patients without pre-existing disorders were followed up. The records of patients who had a cardiovascular event or died were reviewed without knowledge of the patient's sodium-sensitivity status. FINDINGS: 62 patients were deemed sodium sensitive and 94 non-sodium sensitive. Left-ventricular hypertrophy was found more frequently in the sodium-sensitive group than in the non-sodium-sensitive group (38 vs 16%; p < 0.01), whereas significantly fewer patients in this group smoked (23 vs 42%; p < 0.05). There were 17 cardiovascular events in the sodium-sensitive group and 14 in the non sodium-sensitive group. The rate of total, non-fatal and fatal cardiovascular events, was 2.0 per 100 patient-years in the non-sodium-sensitive group and 4.3 per 100 patient-years in the sodium-sensitive group. Cox's proportional-hazards model identified sodium sensitivity (p < 0.01), mean arterial pressure (p < 0.01), and smoking (p < 0.01) as independent cardiovascular risk factors. INTERPRETATION: Cardiovascular events occurred more frequently in patients with sodium-sensitive hypertension. Sodium sensitivity is an independent cardiovascular risk factor in Japanese patients with essential hypertension. PMID- 9413465 TI - Nosocomial transmission of Mycobacterium bovis resistant to 11 drugs in people with advanced HIV-1 infection. AB - BACKGROUND: Since 1990, several nosocomial outbreaks of multidrug-resistant (MDR) tuberculosis have occurred, none of which have involved Mycobacterium bovis. We describe an epidemic of nosocomial and primary MDR M bovis tuberculosis from December, 1993, to February, 1995, among HIV-1-infected patients in a district of Madrid. METHODS: We undertook genetic characterisation of the M bovis strain and investigated its presence in a tuberculosis epidemic in a Madrid hospital in a case-controlled study. We assessed 19 cases diagnosed with MDR tuberculosis due to M bovis during the study period. For the control group, we randomly selected 33 patients with HIV-1 infection and isolation of a strain of M tuberculosis susceptible to isoniazid, rifampicin, or both, who were treated in Ramon y Cajal Hospital. Infection-control policies and practices were implemented. FINDINGS: We detected 19 cases in HIV-1-infected patients of primary MDR tuberculosis produced by M bovis resistant to 11 antituberculosis drugs. We found phenotypic and genotypic similarities in the strains of M bovis. In the case group, the index case and two other cases had had previous contact with another hospital that had had an MDR tuberculosis outbreak. All patients died after a mean of 44 days (range 2-116), despite multidrug treatment with first-line and second-line antituberculosis drugs. The cases with M bovis MDR tuberculosis were significantly more likely than controls to have been admitted to a hospital ward at the same time as patients already infected with MDR tuberculosis during the 10 months before their diagnosis (adjusted odds ratio 94.6 [95% CI 9.4-956.3], p < 0.0001). Advanced HIV-1 immunosuppression was associated with the development of MDR tuberculosis. Implementation of control measures stopped the epidemic. INTERPRETATION: An M bovis primary MDR tuberculosis epidemic that cannot be treated effectively and with high mortality has emerged in Europe and has been transmitted between hospitals. PMID- 9413466 TI - Effect of danaparoid sodium on hard exudates in diabetic retinopathy. AB - BACKGROUND: In diabetes mellitus, both retinopathy and nephropathy represent specific microvascular disease with increased capillary permeability resulting in hard exudates, foveal oedema, and albuminuria. The decrease of heparan-sulphate content of the glomerular-basement membrane is quantitatively related to the rate of proteinuria in nephropathy associated with insulin-dependent diabetes mellitus (IDDM). Several short-term studies in patients with IDDM and non-IDDM have shown that a reduction of microalbuminuria and macroalbuminuria can be achieved with the supplementation of glycosaminoglycans. After completion of a study on the effect of danaparoid sodium on albumin excretion in patients with IDDM and macroalbuminuria, we hypothesised that treatment with danaparoid sodium also influenced retinal leakage in the patients in that trial. METHODS: In this retrospective study nine patients with nephropathy received 750 anti-Xa units danaparoid sodium once a day for 6 weeks in a placebo-controlled double-blind cross-over study. Fundus photographs, done at baseline and at the end of the study, were semiquantitatively scored for the severity of hard exudates. FINDINGS: At baseline 14 eyes had grade 1 to 5 severity of hard exudates and four eyes were without hard exudates (grade 0). There was no progression in the latter four eyes. In ten eyes an improvement was observed: four patients showed a favourable response to treatment in both eyes and two patients showed improvement in one eye. We found improvement of hard exudates after 6 weeks of treatment with danaparoid sodium. INTERPRETATION: Our uncontrolled observation indicates that the supplementation of danaparoid sodium influences both the permeability of retinal vessels as well as of glomerular vessels. Danaparoid-sodium therapy as a systemic adjuvant is worth considering for treatment strategies for foveal oedema and hard exudates in diabetic maculopathy. PMID- 9413467 TI - Unwelcome orgasms. PMID- 9413468 TI - Efficacy of repeated intravenous immunoglobulin in severe unresponsive Guillain Barre syndrome. PMID- 9413469 TI - Mycophenolate mofetil and cyclosporin treatment for recalcitrant pyoderma gangrenosum. PMID- 9413470 TI - Increase of lipoprotein (a) with troglitazone. PMID- 9413471 TI - Procalcitonin aids diagnosis of adrenocortical failure. PMID- 9413472 TI - Expression of natural peptide antibiotics in human skin. PMID- 9413473 TI - Management of pelvic floor dysfunction. PMID- 9413474 TI - Loss of aura in lamotrigine-treated epilepsy. PMID- 9413475 TI - Half-life of truth in surgical literature. PMID- 9413476 TI - HIV-1 infection puzzles Denmark. PMID- 9413477 TI - Antiretrovirals planned for developing world. PMID- 9413478 TI - US admits defeat on HIV drugs. PMID- 9413479 TI - The autistic spectrum. PMID- 9413480 TI - Future vaccines and a global perspective. AB - Advances in medical biotechnology mean that vaccines to prevent more than 75 infectious diseases are being or have been developed. Vaccination is unfortunately not reliant purely on biotechnology but also on politics and resources. Countries with the greatest demand for vaccines have the least ability to pay for or produce them. Health-care Infrastructure and diagnostic facilities also hamper immunisation projects in developing countries. Charitable organisations are relied on heavily to support such projects but the challenge to ensure all infants are immunised against the most common infections of childhood is still enormous. Difficulties that present themselves now should not prevent us looking into future possibilities such as immunisation during pregnancy and targeting of children for immunisation against sexually transmitted diseases. Other avenues for research are in administration of vaccines. A move to mucosal immunisation rather than use of the syringe and needle would be positive both economically and from the point of view of risk of needle contamination. Plant science may also provide a new vehicle for vaccines by engineering plants such as the banana tree to be naturally bioencapsulated vaccines. Prospects for control and eradication of infectious disease in the next century are certainly good. PMID- 9413481 TI - The disease of Immanuel Kant. PMID- 9413482 TI - Measles vaccination and inflammatory bowel disease. PMID- 9413483 TI - Measles vaccination and inflammatory bowel disease. PMID- 9413484 TI - Measles vaccination and inflammatory bowel disease. PMID- 9413485 TI - Effect of paracetamol on parasite clearance time in Plasmodium falciparum malaria. PMID- 9413486 TI - Cutaneous T-cell lymphoma severity index and T-cell gene rearrangement. PMID- 9413487 TI - Molecular genetic tests in surgical management of familial adenomatous polyposis. PMID- 9413488 TI - Stroke after visit to the hairdresser. PMID- 9413489 TI - Total cholesterol and mortality after age 80 years. PMID- 9413490 TI - Disability in the elderly. PMID- 9413491 TI - Older patients and advances in cardiology. PMID- 9413492 TI - HIV-1 infection associated with abnormal vaginal flora morphology and bacterial vaginosis. PMID- 9413493 TI - Thromboembolic risk and pacemakers. PMID- 9413494 TI - From efficacy to cost-effectiveness. PMID- 9413495 TI - From efficacy to cost-effectiveness. PMID- 9413496 TI - Constrictive pericarditis and tuberculosis. PMID- 9413497 TI - Constrictive pericarditis and tuberculosis. PMID- 9413498 TI - Antenatal HIV testing in Europe. PMID- 9413499 TI - Hazards of powdered surgical gloves. PMID- 9413500 TI - Big programmes, big errors? PMID- 9413501 TI - Pharmaceutical representatives. PMID- 9413502 TI - Psion of the times. PMID- 9413503 TI - Isolation of a measles virus variant: protection of newborn mice from measles encephalitis by 24 h prior intracerebral inoculation with the variant. AB - A small plaque mutant with reduced neurovirulence in newborn mice was obtained from Edmonston strain measles virus after propagation for 5 months in NIH3T3 cells. It retained the antigenicity of the parental virus and tended to induce higher neutralizing antibody titers in the adult BALB/c mice. The intracerebral (but not intraperitoneal) inoculation of the live mutant virus one day before prevented the newborn BALB/c mice from encephalitis caused by the intracerebral challenge with the parental strain at a dose of 10-20 LD50. The intracerebral inoculation with the mutant virus whose replication capacity was inactivated by UV-irradiation was ineffective. The protection was not attributed to interferons nor to viral interference. The mechanism remains unknown. PMID- 9413504 TI - Populations of two eastern countries of Japan and Korea and with a related history share a predominant genotype of herpes simplex virus type 1. AB - Herpes simplex virus type 1 (HSV-1), a common human pathogen of non-epidemic nature is linked closely to the individual by latent infection. HSV-1 genotypes usually differ with race. Based on a "dual structure model" for population history of the Japanese, modern Japanese populations are assumed to have derived from two major migration events. The Jomon people arrived in Japan > 10,000 years ago and the Yayoi people began migrating to Japan from the Korean peninsula approximately 2,300 years ago. The presence of two predominant genotypes of F1 and F35 was noted in HSV-1 strains isolated in Japan. As the F1 genotype also predominated in Korea, peoples of Japan and Korea share the F1 genotype. Regional differences in the frequency of F1 and F35 genotypes within Japan seem to relate to differences in the dispersion of descendants of the Yayoi and Jomon peoples. Our hypothesis is that the F35 genotype relates to the Jomon people, earlier residents in japan, while the F1 genotype relates to the Yayoi people who migrated much later from the Korean peninsula. PMID- 9413505 TI - Comparison of gelatin particle agglutination and hemagglutination inhibition tests for measles seroepidemiology studies. AB - The prevalence of measles antibody in Japan was surveyed with a newly developed gelatin particle agglutination (PA) test, and the results compared with those of the hemagglutination inhibition (HI) test. The two age-distribution curves of the PA antibody-positive rates at > or = 1:8 and > or = 1:32 were almost the same in all the age groups, except the less-than-1-year-old group for which the rate at > or = 1:8 was higher than that at > or = 1:32 (p < 0.05, chi 2 test). In the vaccinated children, all groups older-than-1-year of age had antibody-positive levels of 96% or more. In contrast, in the unvaccinated children, there was a sharp increase in antibody-positive rates between the 1- and 4-year-old groups, indicative that about 80% of the children were infected by wild measles virus at these ages. A significant number of PA antibody-positive specimens were antibody negative (< 1:8) by HI. The percentage of specimens in this category, PA (+) but HI (-), was greatest in infants less than one year old, and least in young children, but it increased with age to 97% of the HI (-) specimens from adults of more than 20 years of age. The PA test therefore detected some measles antibodies that HI could not. This test is simple and useful for making serosurveys in both developed and developing countries. PMID- 9413506 TI - Development of a gelatin particle agglutination reagent for measles antibody assay. AB - A new agglutination test that utilizes gelatin particles as the carrier of measles antigen was developed and used to evaluate immune status to measles virus infection. The particle agglutination (PA) reagent reacted with monoclonal antibodies against two major proteins of measles virus, the hemagglutinin (H) and fusion (F) proteins. Children were followed individually for ten years for measles antibody. Results showed that the PA test was as sensitive and specific as the plaque neutralization test. The procedure is simple and rapid. No prior treatment of specimens is needed, and the test is completed in a single reaction. The PA test therefore can be used for diagnoses and epidemiologic surveys of measles virus infection. PMID- 9413507 TI - Phylogenetic analysis of the Potyviridae with emphasis on legume-infecting potyviruses. AB - The 3'-terminal nucleotide sequences of thirteen authenticated strains of bean common mosaic virus (BCMV) and one strain of bean common mosaic necrosis virus (BCMNV) were obtained. The regions sequenced included the coat protein coding sequence and 3'-end non-coding region. These data, combined with sequence information from other legume-infecting potyviruses and the Potyviridae were used for phylogenetic analysis. Evidence is provided for delineation of BCMNV as distinct from BCMV and the inclusion of azuki mosaic, dendrobium mosaic, blackeye cowpea mosaic, and peanut stripe viruses as strains of BCMV. This relationship defines the members of the BCMV and BCMNV subgroups. These data also provide a basis upon which to define virus strains, in combination with biological data. Other aspects and implications of legume-infecting potyvirus phylogenetics are discussed. PMID- 9413508 TI - Anti-MHV3 state induced by IFN gamma in macrophages is not related to arginine metabolism. AB - In contrast to BALB/c mouse macrophages, the A/J macrophages after activation by interferon gamma (IFN gamma) develop an anti-MHV3 effect which correlates with the resistance to virus infection. To understand the cellular basis of this antiviral effect, we studied the possible involvement of arginine metabolism through nitric oxide (NO) and arginase induction, since these metabolic pathways have been described as implicated in antiviral activities of macrophages. The studies were performed by activating macrophages with inducers of NO (IFN gamma) and arginase (IL4 IL10). NO synthase (iNOS) and arginase inhibitors (N-methyl arginine, NMA, and hydroxyarginine, OH-ARG) were used. The results show that in both macrophage populations, no spontaneous synthesis of NO occurred and the MHV3 enhanced the NO release induced by IFN gamma. After activation with IFN gamma, BALB/c macrophages released higher amounts of NO than the A/J macrophages. The inhibition of IFN gamma-induced NO-synthesis with NMA or with arginine free medium did not affect the virus replication. In BALB/c macrophages, IL4 or IL10, induced higher amounts of arginase than in A/J macrophages. In both macrophage populations the MHV3 infection had no influence on the arginase synthesized, and the inhibition of the arginase with OH-ARG had no influence on the virus growth. The level of MHV3 replication or inhibition was also not influenced when we used macrophages from knockout mice for the iNOS gene, and as a consequence were unable of synthesizing NO. These data indicate that NO and arginase do not participate in the anti-MHV3 state induced by IFN gamma in macrophages. PMID- 9413509 TI - The minor outer capsid protein P2 of rice gall dwarf virus has a primary structure conserved with, yet is chemically dissimilar to, rice dwarf virus P2, a protein associated with virus infectivity. AB - The nucleotide sequence of the genome segment 2 (S2) of rice gall dwarf virus (RGDV), a phytoreovirus, when compared with the amino acid sequence of a component protein of the virus, showed that S2 potentially encoded a 127K minor outer capsid protein. This 127K protein designated as P2 and the 127K minor outer capsid protein (also termed P2) of rice dwarf virus (RDV) are similar in size, located in the outer capsid, and have well-conserved predicted polypeptide sequences, suggesting similar functions. Infectivity to insect vector cell monolayers of RGDV was maintained and the P2 protein was retained irrespective of carbon tetrachloride (CCl4) treatment. This is in contrast to the infectivity of RDV which is removed along with P2 protein following CCl4 treatment. RGDV with P2 was acquired by vector insects and transmitted to host plants, although RDV lacking P2 could not be transmitted to plants as previously published. These results imply that RDV and RGDV require P2 proteins for virus infectivity to vector insects. PMID- 9413511 TI - Amantadine does not have antiviral activity against Borna disease virus. AB - We have investigated the antiviral activity of amantadine (AD) against Borna disease virus (BDV) in several culture cell systems. We present evidence that AD, in the range 5 to 10 microM, does not have antiviral activity against BDV. Treatment of BDV infected cells with AD for six days caused neither a reduction in the number of infected cells, nor a decrease in steady state levels of BDV RNA or proteins. Moreover, treatment of cells with AD prior infection did not affect BDV multiplication, whereas influenza A virus yield was less than 1% with respect to that obtained in untreated control cells. PMID- 9413510 TI - The non-structural polyprotein 3ABC of foot-and-mouth disease virus as a diagnostic antigen in ELISA to differentiate infected from vaccinated cattle. AB - A diagnostic assay to differentiate antibodies induced by foot-and-mouth disease virus (FMDV) infection from those induced by vaccination was developed. The test is an indirect-trapping ELISA which uses a monoclonal antibody to trap the non structural 3ABC-FMDV polypeptide expressed in E. coli. Experimental and field sera from naive, vaccinated and infected cattle were examined. Using the established threshold of 0.20 optical density units, the sensitivity of the assay was 100%, as all the experimental post-infection sera (n degree = 137) gave values greater than this threshold, irrespective of the FMDV serotype used for the infection. In contrast, more than 99% of sera from vaccinated animals were negative (225 out of 228 primo-vaccinates and 159 out of 159 multi-vaccinates). A high degree of specificity was also confirmed by the finding that 99.5% (442 out of 444) of sera from naive animals gave negative results. Serum conversion against 3ABC was first detected 8 days post-infection and demonstrable levels of 3ABC specific antibodies were detectable at least 1 year post-infection. The described 3ABC-ELISA is safe, cheap and also easy to perform in large scale serological surveys. The high specificity and sensitivity makes this test an ideal tool for FMD eradication campaigns and control programs. PMID- 9413512 TI - Borna disease virus is not sensitive to amantadine. AB - Successful inhibition of Borna disease virus (BDV) by amantadine in cultured cells and in an infected human individual has been reported [Bode et al. (1997) Lancet 349: 178-179]. We now found that infection of monkey Vero cells by laboratory strains of BDV was not influenced by amantadine under conditions that reduced the yields of influenza A virus by about 400 fold. Amantadine treatment of Vero cells persistently infected with BDV did not result in reduced viral RNA levels, and application of the drug to persistently infected BALB/c mice had no effect on the concentration of BDV in their brains. Thus, susceptibility to amantadine is not a characteristic of BDV, although it may be observed with certain primary virus isolates. PMID- 9413513 TI - Genomic (5'UTR) and serological differences among German BVDV field isolates. AB - Isolates of bovine viral diarrhoea virus (BVDV) collected in Germany were examined for their genomic heterogeneity in sequences from the 5'untranslated region (UTR) of the viral genome. Polymerase chain reaction (PCR) tests based on the 5'UTR and the region coding for the NS2-3/4A polypeptide were used to differentiate between BVDV I and BVDV II genotypes. Eleven out of 96 BVDV isolates were identified as BVDV II. Virus neutralization tests with BVDV I- or II-specific antisera raised in cattle were done. The mean titers were reduced by 7.2-fold (BVDV I-antiserum versus type II-isolates) or 35-fold (BVDV II-antiserum versus type I-isolates) when using the respective heterologous virus. PMID- 9413514 TI - Analysis of the non-sense mutants of varicella-zoster virus thymidine kinase. AB - Three non-sense mutants of varicella-zoster virus (VZV) thymidine kinase (TK) gene, VZTK325, VZTK278 and VZTK224, were isolated. The mutants had a single nucleotide substitution at codons 326, 279 and 225, which changed the codon of TGG (tryptophan) to the stop codon TGA. The wild type (WT) and mutant TKs were expressed in E. coli cells and their characteristics were evaluated. VZTK224 lost TK activity, but VZTK325 and VZTK278 maintained 74.8% and 21.2% of the WT TK activity. On the other hand, all mutants lost the thymidylate kinase activity. Moreover, VZTK325 and VZTK278 polypeptides were heat-labile. These data suggest that the carboxy-terminal portion of herpesvirus TK plays an important role in the stable folding of TK and thymidylate kinase activity. PMID- 9413515 TI - Distribution of VP7 serotypes and VP4 genotypes among rotavirus strains recovered from Italian children with diarrhea. AB - 108 rotavirus strains obtained from children with diarrhea hospitalized in Palermo, Italy, in the years 1990-1994, were examined by seminested PCR to study the relative frequency and distribution of the four most common alleles of the gene 4. Such strains were selected from 344 human rotavirus strains recovered in palermo during those years after characterization by electropherotyping, subgrouping and G serotyping. One hundred and seven of the 108 strains could be classified into P types, the P[8], G1 (38.3%) and the P[8], G4 (52.3%) types being predominant. The unique strain whose P genotype could not be identified showed an unusual combination of long migration electrophoretic pattern and subgroup I specificity. PMID- 9413516 TI - Electron microscopic observation on a non-occluded baculo-like virus in shrimps. AB - A severe disease of farm-raised shrimp, Penaeus chinensis has spread throughout Chinese coasts since 1993. Recently, a baculo-like virus has been diagnosed as the causative agent for this epidemic disease. The electron microscopic observation of the virus is described. Thin sections of hepatopancreatic and hypodermic tissue of diseased P. chinensis showed many rod-shaped, enveloped, baculo-like virions in hypertrophied nuclei of infected cells. The virion was filled with a highly electron dense core. No occlusion bodies have been found. Negative stained intact virions, purified from infected tissues by centrifugation on sucrose discontinuous gradients, demonstrated that the viral envelops had been broken, but the cylindrical nucleocapside could be observed clearly. The nucleocapsid of average 62 nm x 314 nm was composed of a helix system of capsomers, giving rise to an open stacked ring structure and repeating approximately every third ring. The number of repeating unit was 13 to 15. We propose that the virus described here could be designated as Non-Occluded Shrimp Virus (NOSV) or Penaeus chinensis Baculo-Like Virus (PcBLV). PMID- 9413517 TI - Limited antigenic variation among recent infectious bursal disease virus isolates from France. AB - Seven neutralizing monoclonal antibodies (Mabs) to infectious bursal disease virus (IBDV) were used in an antigen-capture ELISA for the antigenic characterization of 58 IBDV isolates obtained in France since 1989. Fifty-six isolates exhibited an antigenic profile which was different from reference strain Faragher 52/70, and similar to French very virulent IBDV strain 89,163 (no binding of two Mabs). Two strains (3.4%), isolated in 1991 and 1994, showed additional antigenic modifications resulting in suppressed or reduced binding activity of three other Mabs. The two atypical viruses were not re-identified in field samples subsequently collected, hence showing that antigenic variants of IBDV do not tend to replace the antigenically dominant 89,163-like strains that have prevailed in France since 1989. PMID- 9413518 TI - Synthetic peptides from the heptad repeats of the glycoproteins of rabies, vesicular stomatitis and fish rhabdoviruses bind phosphatidylserine. AB - This work follows up on observations previously published concerning phosphatidylserine (PS) binding properties of synthetic peptides (p2) from the hydrophobic heptad repeats of the glycoprotein of viral haemorrhagic septicemia (VHS) rhabdovirus and the presence of similar repeats in the sequences of the glycoproteins of four separate rhabdoviruses. Similar p2-like peptides are now synthesized according to the corresponding cDNA sequences of infectious haematopoietic necrosis (IHN), rabies and vesicular stomatitis (VSV) viruses and shown to bind phosphatidylserine (PS) by solid-phase as well as from liquid-phase assays. The PS-binding peptides located in the amino-terminal part of the glycoproteins contained 3-5 contiguous heptad repeats (abcdefg) of hydrophobic amino acids (aa) in positions a and d followed by a short aa stretch containing positively charged aa and not belonging to the heptad repeats. The rhabdoviral PS binding regions had low sequence variability among the members of each of the rhabdoviral genus but show no sequence similarity among the different genera. PMID- 9413519 TI - Stimulation of type I DNA topoisomerase gene expression by pseudorabies virus. AB - Previous results from our laboratory have demonstrated that type I DNA topoisomerase activity is required for the replication and gene expression of pseudorabies virus (PRV). In the present report, we further analyzed the expression of topoisomerase I in PRV-infected cells, and the western blot result showed that the expression of topoisomerase I was increased after virus infection. The increase sustained to late time of infection when the cytopathic effect was obvious and the synthesis of most host proteins was shut off by PRV. From transient expression assay, it was also found that the promoter of cellular topoisomerase I gene could be stimulated by immediate-early protein (IE180) and viral early protein 0 (EP0), and these two regulatory proteins appeared to work synergistically. Collectively, these findings provide evidence that PRV can stimulate the expression of topoisomerase I and that the stimulation is mediated at least by IE180 and EP0 proteins of PRV at the transcriptional level. PMID- 9413520 TI - Evidence for rolling circle replication of Autographa californica M nucleopolyhedrovirus genomic DNA. AB - Autographa californica M nucleopolyhedrovirus (AcMNPV) is a large ds DNA virus restricted to larval lepidopteran insect hosts. Using field inversion gel electrophoresis and digestion with a restriction enzyme which cuts the AcMNPV genome once, we detected multiple unit-length genome fragments from replicating viral DNA. Our data suggest that AcMNPV replicates in a head-to-tail manner via rolling circle replication. PMID- 9413521 TI - Cowpox: a re-evaluation of the risks of human cowpox based on new epidemiological information. AB - Human cowpox is a rare but relatively severe infection of interest because of its links with Edward Jenner and the introduction of smallpox vaccine and, more recently, because of re-evaluation of the epidemiology of the infection. This indicates that cowpox is not enzootic in cattle, relegates the cow to a minor role, and emphasizes the importance of feline cowpox as a source of human infection and of wildlife as virus reservoirs. The evidence available suggests that the virus is of low infectivity for humans and should not become an increasing problem despite the cessation of smallpox vaccination and increasing numbers of immunocompromised individuals. PMID- 9413522 TI - Characterization of a cowpox-like orthopox virus which had caused a lethal infection in man. AB - In August 1990 an orthopox virus (OPV) had been isolated from a severe case of a generalized infection with lethal outcome in an immunosuppressed 18-year-old man. In this communication we present a detailed characterization of the causative virus strain. Based on distinct epitope configurations detected by various monoclonal antibodies the isolate could be differentiated from other OPV species and was classified as a cowpox virus (CP). This classification was confirmed by a species-specific PCR assay and by establishing physical maps for the restriction enzymes HindIII and XhoI. Based on serological data of neutralization assays, blocking-ELISAs and Western blotting analysis evidence is provided that the infection had been acquired from a stray cat. PMID- 9413523 TI - Molecular genetic analyses of parapoxviruses pathogenic for humans. AB - The current members of the genus parapoxvirus are orf virus (ORFV), bovine papular stomatitis virus (BPSV), pseudocowpoxvirus (PCPV) and parapoxvirus of red deer in New Zealand (PVNZ). BPSV and PCPV are maintained in cattle while ORFV is maintained in sheep and goats, but all three are zoonoses. Only the recently reported PVNZ has yet to be recorded as infecting humans. Tentative members of the genus are camel contagious ecthyma virus, chamois contagious ecthyma virus and sealpoxvirus. The separation of the parapoxviruses into 4 distinct groups has been based on natural host range, pathology and, more recently, on restriction endonuclease and DNA/DNA hybridisation analyses. The latter studies have shown that the parapoxviruses share extensive homology between central regions of their genomes, but much lower levels of relatedness within the genome termini. The high G + C content of parapoxvirus DNA is in contrast to most other poxviruses and suggests that a significant genetic divergence from other genera of this family has occurred. DNA sequencing of portions of the genome of ORFV, the type species of the genus, has allowed a detailed comparison with the fully sequenced genome of the orthopoxvirus, vaccinia virus (VACV). These studies have provided a genetic map of ORFV and revealed a central core of 88 kbp within which the genomic content was strikingly similar to that of VACV. This conservation is not maintained in the genome termini where insertions, deletions and translocations have occurred. The characterisation of specific ORFV genes may lead to the construction of attenuated vaccine strains in which genes such as those with the potential to interfere with the immune response of the host have been deleted. The current ORFV vaccines are living unattenuated virus and vaccination lesions produce virus which contaminates the environment in a manner similar to natural infection. The virus in scab material is relatively resistant to inactivation and this virus both perpetuates the disease in sheep and provides the most likely source of human infections. A vaccine which immunises animals without perpetuating the disease could be the best way of reducing the incidence of ORFV infection of humans. It is likely that protection against infection by ORFV is cell mediated and will require the endogenous production of relevant antigens. We have recently constructed a series of VACV recombinants each of which contains a large multigene fragment of ORFV DNA. Together the recombinants represent essentially all of the ORFV genome in an overlapping manner. Vaccination of sheep with the recombinant library provided protection against challenge with virulent ORFV. Further studies with this library may enable dominant protective antigens of ORFV to be identified and lead to their incorporation into a subunit vaccine. PMID- 9413524 TI - Recent advances in molluscum contagiosum virus research. AB - Molluscum contagiosum virus (MCV) and variola virus (VAR) are the only two poxviruses that are specific for man. MCV causes skin tumors in humans and primarily in children and immunocompromised individuals. MCV is unable to replicate in tissue culture cells or animals. Recently, the DNA sequence of the 190 kbp MCV genome was reported by Senkevich et al. MCV was predicted to encode 163 proteins of which 103 were clearly related to those of smallpox virus. In contrast, it was found that MCV lacks 83 genes of VAR, including those involved in the suppression of the host response to infection, nucleotide biosynthesis, and cell proliferation. However, MCV possesses 59 genes predicted to code for novel proteins including MHC-class I, chemokine and glutathione peroxidase homologs not found in other poxviruses. The MCV genomic data allow the investigation of novel host defense mechanisms and provide new possibilities for the development of therapeutics for treatment and prevention of the MCV infection. PMID- 9413525 TI - Molecular anatomy of lymphocystis disease virus. AB - Lymphocystis disease (LD) has been reported to occur in over one hundred different species of fish worldwide. The disease is caused by lymphocystis disease virus (LCDV), a member of the iridovirus family. Numerous fish species that play an important role in fishery and fish farming are highly susceptible to LCDV infection. The infected animals develop disseminated clusters of aberrant hypertrophied cells within their connective tissue, the so-called lymphocystis cells. In the cytoplasm of these cells a massive accumulation of virions can be observed. As a first step towards understanding the mechanisms of viral infection and pathogenesis the complete genomic nucleotide sequence of lymphocystis disease virus type 1 (LCDV-1; flounder isolate) was determined. LCDV-1 is the type species of the genus Lymphocystivirus within the family Iridoviridae. The virions contain a single linear double-stranded DNA molecule that is circularly permuted, terminally redundant and heavily methylated. Since there is no convenient cell system for virus replication we determined the complete nucleotide sequence of the viral genome (102,653 base pairs). Computer assisted analyses of 195 potential open reading frames resulted in the identification of a number of putative gene products with significant homology to functionally characterized proteins of other species. PMID- 9413526 TI - Detection of virus or virus specific nucleic acid in foodstuff or bioproducts- hazards and risk assessment. AB - There are two possibilities for virus contamination of foodstuff and bioproducts of animal origin: i) the presence of endogenous virus as a result of an acute or subclinical infection of animal raw material used for food processing or ii) contamination of food in the course of processing or thereafter. The latter must be considered as the highest risk for human consumers since the viral contamination mostly is caused by virus shedding people and the transmitted viruses are obligate human pathogens. Food from animals consumed as raw material (e.g. oysters) is listed in a high risk category concerning viral contamination (e.g. hepatovirus). Virus contamination of bioproducts such as vaccines, blood products or biological material used in surgery and for transplantations also is more hazardous because the application of contaminating virus usually occurs by circumvention of the natural barrier systems of the body. Moreover, in many cases immunosuppressed people are treated with bioproducts. Due to an enclosing shield of high protein and lipid content in food and bioproducts viruses are well protected against physical and chemical influences, however most preparation procedures for food are destructive for viruses. The detection of pseudorabies virus and pestivirus in biological fluids was tested using polymerase chain reaction (PCR), reverse transcriptase (RT)-PCR and cell culture propagation. PCR is a powerful method to detect viral nucleic acid whereas the detection of infectious virus in cell cultures is more limited, e.g. due to protein and lipid destroying conditions. Virus contamination of bioproducts should be considered as a hazard no matter which method has been used for its detection. Examples are given about the contamination of cell lines and vaccines. PMID- 9413527 TI - Rapid molecular detection of microbial pathogens: breakthroughs and challenges. AB - Microbiological contamination of foods and drinking water is a global problem, and a significant amount of expense is being incurred as a result of such contamination. The microorganisms associated with almost half of all disease outbreaks still go unidentified, primarily as a result of inadequate monitoring and surveillance. Though significant improvements have been made in refining molecular methods for detecting infectious agents, a majority of these methods are being employed only on clinical samples where pathogen densities are much higher than those found in environmental and food samples. Comparative evaluations of the various protocols in terms of cost, sensitivity, specificity, speed, and reproducibility need to be undertaken so that the true applicability of these methods can determined. In the future, molecular methods, especially gene amplifications and in situ hybridizations, will find increasing applications in the differentiation of viable and non-viable organisms, in predicting antimicrobial resistance, and in the identification and characterization of unculturable microorganisms. Though molecular detection methods will not totally replace conventional methods, they will significantly enhance our ability to detect microbial pathogens rapidly. PMID- 9413528 TI - Where do we stand with oral vaccination of foxes against rabies in Europe? AB - The oral vaccination of wild animals was first attempted in 1962 after the repeated failure of poisoning or trapping to control movement of the disease in these species. Foxes were chosen for research purposes because they are a problem animal species and are exquisitely susceptible to rabies. The first successful laboratory studies with attenuated vaccine came in 1971, and the first successful field trial was carried out in Switzerland beginning in 1978. In the 1980's several European countries joined the trials. In the following years many improvements were made: the chicken head was replaced by machine-made baits for easy mass production, the hand placement of vaccine baits was to a greater extent being replaced by aerial distribution (small aircraft or helicopter), and several new vaccines were developed. Additionally, the European Union supported the oral vaccination financially. There was a great impact on the rabies situation. When the second country, Germany, joined the field trial in 1983 the total of reported rabies cases in Europe amounted to 23,002, in 1995 a total of 8,134 cases was reported. In spite of the great improvement made in the past years, in the beginning of the 1990's several severe set-backs were experienced. The paper elaborates on reasons for these set-backs and suggests a strategy to overcome them. PMID- 9413529 TI - Foot- and-mouth disease as zoonosis. AB - Man's susceptibility to the virus of foot- and-mouth disease (FMD) was debated for many years. Today the virus has been isolated and typed (type O, followed by type C and rarely A) in more than 40 human cases. So no doubt remains that FMD is a zoonosis. Considering the high incidence of the disease (in animals) in the past and in some areas up to date, occurrence in man is quite rare. In the past when FMD was endemic in Central Europe many cases of diseases in man showing vesicles in the mouth or on the hands and feet were called FMD. The first suggestion of a human infection with FMD was reported in 1695 by Valentini in Germany [7]. All reports before 1897, the year of the discovery of the virus of FMD by Loeffler and Frosch [2], were not of course confirmed either by isolation of the virus or by identification of immunoglobulins after infection. Nevertheless the successful self-infection reported by Hertwig in 1834 most likely seems to have been FMD in man: each of three veterinarians drank 250 ml of milk from infected cows on four consecutive days. The three men developed clinical manifestations. The diseases most often confused with FMD are infections with several viruses of the Coxsackie A group (this infection is referred to as "hand and mouth disease"), herpes simplex and sometimes vesicular stomatitis. Beginning in 1921 up to 1969 at least 38 papers were published, which described clinically manifest FMD in man in more than 40 proven cases. One further reported described an asymptomatic infection with FMD in man [10]. Criteria for establishing a diagnosis of FMD in man are the isolation of the virus from the patient and/or identification of specific antibodies after infection. Laboratory tests for diagnosis of human FMD are the same as for animals. Proven cases of FMD in man have occurred in several countries in Europe, Africa and South America. The type of virus most frequently isolated man is type O followed by type C and rarely A. The incubation period in man, although somewhat variable, has not been found to be less than two days and rarely more than six days. PMID- 9413530 TI - Molecular epidemiology of influenza. AB - The genome of the influenza A viruses comprises eight single-stranded RNA segments, and this property makes genetic reassortment after double infection of a host with two different influenza A strains possible. Nature takes advantage of genetic reassortment during antigenic shift creating new pandemic strains. After concurrent infection of a host with both avian and human strains, the hemagglutinin gene of the human virus may be replaced by the allelic gene of the avian virus. This reassortment leads to a human virus strain that has avian hemagglutinin molecules on its surface, significant because the human population lacks neutralizing antibodies to this new glycoprotein. The Hong Kong pandemic of 1968 resulted from just such an event. PMID- 9413531 TI - Influenza virus: transmission between species and relevance to emergence of the next human pandemic. AB - Although influenza viruses are not spread from human to human through the conventional food chain, this is not necessarily the case for the transmission of the precursors of the human pandemic influenza viruses. Aquatic birds of the world are the reservoirs for all influenza A viruses; the virus is spread by fecal-oral transmission in untreated water. Influenza A viruses are frequently transmitted to domestic poultry and two of the 15 subtypes H5 and H7 can become highly pathogenic and have the capacity to decimate commercial poultry flocks. Less frequently, avian influenza viruses are transmitted between species-to pigs, horses and sea mammals. This transmission involves mutational, reassortant or recombinational events and can occur through fecal contamination of unprocessed avian protein or through the water. The transmission of avian influenza viruses or virus genes to humans is postulated to occur through pigs that act as the intermediate host. This involves either multiple mutational or reassortant events and is believed to occur by airborne transmission. Once avian influenza viruses are established in mammals, they are transmitted from animal to animal by the respiratory airborne route. The transmission of avian influenza virus from their reservoir in wild aquatic birds to domestic poultry and to mammalian species including humans can be prevented by treatment of the water supply and of avian protein sources with disinfectants or by heating. Agricultural authorities have recommended the separation of wild aquatic and domestic poultry and of pig and poultry farming. It is theoretically possible to reduce the possibility of the next pandemic of influenza in humans by changes in agricultural practices so that ducks are separated from pigs and people. PMID- 9413532 TI - Functional chimeric HN glycoproteins derived from Newcastle disease virus and human parainfluenza virus-3. AB - Newcastle disease virus (NDV) is primarily a respiratory tract pathogen of birds, particularly chickens, but it occasionally produces infection in man. Human parainfluenza virus type 3 (hPIV3) is a common respiratory pathogen, particularly in young children. These two viruses gain entry to host cells via direct fusion between the viral envelope and the cell membrane, mediated by the two surface glycoproteins: the hemagglutinin-neuraminidase (HN) and fusion (F) proteins. Promotion of fusion by HN and F requires that they are derived from homologous viruses. We have constructed chimeric proteins composed of domains from heterologous HN proteins. Their ability to bind cellular receptors and to complement the F protein of each virus in the promotion of fusion were evaluated in a transient expression system. The fusion specificity was found to segregate with a segment extending from the middle of the transmembrane anchor to the top of the putative stalk region of the ectodomain. All of the chimeras, in which the globular domain is derived from the NDV HN and various lengths of the stalk region are derived from the hPIV3 HN maintain receptor binding activity, but some have markedly reduced neuraminidase (NA) activity. Decrease in the NA activity of the chimeras correlates with alteration in the antigenic structure of the globular domain. This suggests that the stalk region of the HN spike is important for maintenance of the structure and function of the globular domain of the HN protein spike. PMID- 9413533 TI - Viral factors determining rotavirus pathogenicity. AB - The pathogenicity of rotaviruses depends on multiple viral and host factors. Evidence is presented for the involvement of a number of viral genes (coding for structural and non-structural proteins) in the ability of the virus to cause diarrhoea. Different genes are important in different rotavirus--host systems suggesting that there is no single viral pathogenicity factor. PMID- 9413534 TI - Viral zoonoses and food of animal origin: caliciviruses and human disease. AB - Caliciviruses are important veterinary and human pathogens. The viruses gain their name from characteristic cup-shaped structures seen on the virion surface by negative stain electron microscopy. In humans caliciviruses are a major cause of diarrhoeal disease. There are two fundamentally different genome structures amongst human caliciviruses. The Norwalk-like or small round structured viruses (SRSVs) are viruses that have an amorphous structure when viewed by EM, they have a genome composed of 3 major open reading frames (ORFs). These viruses cause epidemic gastroenteritis amongst all age groups. In contrast, the 'classic' human caliciviruses (HuCVs) display the typical calicivirus surface structure and have their capsid ORF fused to and contiguous with the non structural proteins forming one giant polyprotein. HuCVs are predominantly associated with paediatric infections and are only a minor cause of disease in humans. Spread of disease for both SRSVs and HuCVs is usually by faecal oral transmission. SRSVs are a major cause of foodborne gastroenteritis especially linked to the consumption of sewage contaminated shellfish. However, there is no evidence that these viruses replicate in shellfish or that they originate from an animal source. PMID- 9413535 TI - The role of human caliciviruses in epidemic gastroenteritis. AB - Members of the Caliciviridae family of small, positive-sense RNA viruses exhibit a broad host range. The Norwalk and Norwalk-like caliciviruses in this family are major etiologic agents of epidemic gastroenteritis in humans. This illness characteristically lasts 24-48 h and often occurs in group settings such as families, schools, institutions, or communities. The spread of the human caliciviruses is considered to be predominantly by person-to-person contact via the fecal-oral-route. However, the ingestion of calicivirus-contaminated food or water can result in large-scale common-source outbreaks. Many basic features concerning the biology and replication of the human caliciviruses are not known because they have not yet been grown in cell culture and the virus does not appear to replicate in animal models other than the chimpanzee. Sequence analysis of RT-PCR-generated DNA fragments derived from serotypically distinct reference strains (such as the Norwalk, Hawaii, and Snow Mountain viruses) and other circulating strains associated with gastroenteritis has provided evidence for marked genetic diversity among these viruses. Moreover, analysis of the antigenic relationships among these viruses using paired sera from individuals infected with well-characterized reference strains or from animals immunized with recombinant "virus-like particles" (VLPs) suggests that several serotypes of these viruses are circulating worldwide. The availability of molecular techniques for the detection of these fastidious viruses has enabled epidemiologic studies that have strengthened the association of human caliciviruses with acute gastroenteritis and has demonstrated a potential role for antigenic diversity in the natural history of these pathogens. PMID- 9413536 TI - Clinical similarities and close genetic relationship of human and animal Borna disease virus. AB - Borna disease virus (BDV) is the prototype genus of a new family, Bornaviridae, within the order Mononegavirales. BDV naturally infects animals and man. The symptomatology in animals ranges from subclinical infection to rare cases of encephalitis. Asymptomatic infection seemed more frequent than expected, based on antibody data from 100 healthy horses derived from different stables with a history of diseased cases (30-40% carriers). Likewise, phasic episodes of a neurobehavioral syndrome followed by recovery were much more common than fatal neurologic disease. They were paralleled by expression of BDV antigens (N-protein p40, P-protein p24) and RNA transcripts in peripheral blood mononuclear cells, indicating viral activation. Representative longitudinal studies showed that episodes of depressive illness in humans as well as apathetic phases in infected horses were accompanied by antigen expression and followed a similar clinical course. After recovery, BDV antigen disappeared. This temporal congruence, together with the recent isolation of infectious BDV from such patients, points to a contributory role of this virus in human affective disorders. Successful amelioration of BDV-induced neurobehavioral disease in horses with antidepressants applied in psychiatry, supported a common viral pathomechanism, involving reversible disturbances of the neurotransmitter network in the limbic system. Sequences of genetic material amplified from infected animal tissue and human PBMCs revealed a close interspecies relationship and high sequence conservation of the BDV genome. In human BDV isolates, however, single unique mutations were prominent in four genes. This finding supports the hypothesis that despite of high genomic conservation, species-specific genotypes may be definable, provided the sequences are derived from RNA of infectious virus. PMID- 9413538 TI - Haemorrhagic fevers and ecological perturbations. AB - Hemorrhagic fever is a clinical and imprecise definition for several different diseases. Their main common point is to be zoonoses. These diseases are due to several viruses which belong to different families. The Flaviviridae have been known for the longest time. They include the Amaril virus that causes yellow fever and is transported by mosquitoes. Viruses that have come to light more recently belong to three other families: Arenaviridae, Bunyaviridae, and Filoviridae. They are transmitted by rodents (hantaviruses and arenaviruses) or from unknown reservoirs (Ebola Marburg). The primary cause of most outbreaks of hemorrhagic fever viruses is ecological disruption resulting from human activities. The expansion of the world population perturbs ecosystems that were stable a few decades ago and facilitates contacts with animals carrying viruses pathogenic to humans. Another dangerous human activity is the development of hospitals with poor medical hygiene. Lassa, Crimean-Congo or Ebola outbreaks are mainly nosocomial. There are also natural environmental changes: the emergence of Sin Nombre in the U.S. resulted from heavier than usual rain and snow during spring 1993 in the Four Corners. Biological industries also present risks. In 1967, collection of organs from monkeys allowed the discovery in Marburg of a new family of viruses, the Filoviridae. Hemorrhagic fever viruses are cause for worry, and the avenues to reduce their toll are still limited. PMID- 9413537 TI - Molecular characterization of Borna disease virus from naturally infected animals and possible links to human disorders. AB - In this review data are presented which indicate a high degree of genetic stability of BDV in his natural host, the horse. Despite this high degree of sequence conservation, variation in antigenicity was found, which did not influence the pathogenic properties of the virus. In addition, the correlation between BDV-seropositivity and a variety of psychiatric and neurological disorders in humans is discussed. In diagnostically unselected psychiatric patients we found a similar distribution of psychiatric disorders in BDV seropositives compared to seronegatives. Investigations of cerebrospinal fluid revealed cases of BDV encephalitis in BDV seropositive psychiatric and neurological patients. In contrast to others, we have found no evidence for the presence of BDV-RNA or BDV in human peripheral blood leucocytes. PMID- 9413539 TI - Transmission, species specificity, and pathogenicity of Aujeszky's disease virus. AB - Aujeszky's disease virus (ADV), also known as pseudorabies virus (PrV), is an alphaherpesvirus that causes fatal infections in a wide range of animal species. The virus shares a variety of biological properties with human pathogenic herpesviruses like herpes simplex virus or varicella-zoster virus. Although only limited data are available, it seems unlikely that PrV causes disease in immunocompetent humans, but may pose a risk for immunocompromised patients. PMID- 9413540 TI - The role of veterinary public health in the prevention of zoonoses. AB - Veterinary public health is a component of public health activities devoted to the application of professional veterinary skills, knowledge, and resources for the protection and improvement of public health. VPH activities involve a very diverse range of functions within public health which reflect the broad community of interests between veterinary and human medicine. Zoonoses continue to represent an important health hazard in most parts of the world, where they cause considerable expenditure and losses for the health and agricultural sectors. Although the situation is improving in the industrialized world, zoonoses prevention and control will remain an area of major concern in most developing countries. Recent observations in these countries show that expenses related to the prevention of zoonotic diseases in humans are likely to increase dramatically in the near future. Programmes for their control and eventual elimination in animal reservoirs are urgently needed. The technical knowledge exists to bring diseases such as brucellosis, rabies, and bovine tuberculosis under control during the first decade of the next century. To achieve this goal, constant efforts will be needed for the next 15 to 20 years. In addition, as trade in animal products and the movement of human populations continues to increase, the risk that zoonotic diseases will be introduced or reintroduced into certain areas is likewise increasing. Over the past five years, a number of zoonotic diseases have emerged as either new pathological entities or known agents appearing in new areas or as new strains. Through its coordinating and information gathering functions, the WHO Emerging Disease Surveillance and Control Division provides a source of both practical and technical guidance that can help solve these and other threats to human health posed by animals. PMID- 9413541 TI - Viral infections transmitted by food of animal origin: the present situation in the European Union. AB - The goal of this presentation was to clarify which foods are involved in viral diseases, which viruses are transmitted via food and how to evaluate the risk of a foodborne viral infection. Food items frequently identified as cause of viral disease outbreaks were shellfish harvested in sewage-contaminated water. Another common source of foodborne viral illness was cold food contaminated by infected food handlers. In the European Union the viruses most frequently associated with foodborne illness were hepatitis A virus and the SRSV's. A few isolated cases of foodborne hepatitis E were reported in Mediterranean countries. Compared to other foodborne diseases, those caused by viruses are less severe and seldom fatal. This might be a reason why the problem of viral contamination of food has been neglected. Yet, because many foodborne viral diseases are not recognized either as foodborne or as caused by viruses, the actual number of cases must be assumed to be significantly higher than the reported number. Consequently, food associated diseases of viral origin should be granted more attention. PMID- 9413542 TI - Viral zoonosis from the viewpoint of their epidemiological surveillance: tick borne encephalitis as a model. AB - Tick-borne encephalitis (TBE) is a vector borne and, more rarely, a food (milk, milk products) borne disease of humans. For further characterization of the virus activity in natural foci of TBE more than 32,000 unengorged wild ticks were caught in low and high virus active foci in Germany (Mecklenburg-Western Pomerania, Saxony, Brandenburg, Thuringia, Bavaria, Baden-Wurttemberg, Saarland). The ticks were examined by RT-PCR and Southern blot hybridization as well as by classical virological methods. The dynamics of such natural foci of TBE in the last 35 years were discussed. Also nucleotide sequence data of parts of the virus genome (5'-non coding region) of 16 European and some Far East subtype strains were compared. PMID- 9413543 TI - Strategies to avoid virus transmissions by biopharmaceutic products. AB - The use of biopharmaceutical products offers an opportunity for the treatment of many diseases. Biotechnical manufacturing using recombinant mammalian cell lines is the most appropriate method today for the production of biopharmaceutical protein drugs for the treatment of human and animal diseases. However, mammalian cell line derived products have a potential risk for virus transmission to patients who are treated with these biopharmaceutical products. The reliability that biological products are free of any viruses requires a combination of several strategies: The use of well-characterized cell bank systems and, if feasible, the avoidance of biological raw materials for the cultivation of these mammalian cell lines and the production of biopharmaceuticals. Further on, the purification process for biopharmaceuticals has to be validated for its ability to efficiently remove and inactivate any potential virus contamination and, where applicable, also unconventional transmissible agents, such as BSE. In addition, the biopharmaceutical product itself can be tested for the presence of viruses. Like other manufacturing processes, biotechnical production processes have to be performed in compliance with current Good Manufacturing Practices (cGMP). PMID- 9413544 TI - Comparisons of catalytic selectivity of cytochrome P450 subfamily enzymes from different species. AB - Historically there has been considerable interest in comparing patterns of biotransformation of xenobiotic chemicals in experimental animal models and humans, e.g. in areas such as drug metabolism and chemical carcinogenesis. With the availability of more basic knowledge it has become possible to attribute the oxidation of selected chemicals to individual cytochrome P450 (P450) enzymes in animals and humans. Further, these P450s can be characterized by their classification into distinct subfamilies, which are defined as having > 59% amino acid sequence identity. Questions arise about how similar these enzymes are with regard to structure and function. More practically, how much can be predicted about reaction specificity and catalysis? In order to address these issues, we need to consider not only the relatedness of P450s from different species but also (i) functional similarity within P450 subfamilies and (ii) the effects of small changes imposed by site-directed mutagenesis. Relationships in the P450 1A, 2A, 2B, 2C, 2D, 2E, 3A, and 17A subfamilies are briefly reviewed. Overall functional similarity is generally seen in subfamily enzymes but many examples exist of important changes in catalysis due to very small differences, even a single conservative amino acid substitution. Some general conclusions are presented about predictability within various P450 subfamilies. PMID- 9413545 TI - Antioxidative activity of ginkgolides against superoxide in an aprotic environment. AB - The terpene lactones ginkgolide A, ginkgolide B, ginkgolide C, ginkgolide J and bilobalide, which are components of a standardized extract (EGb 761) from leaves of Ginkgo biloba, as well as ginkgolide M from roots of G. biloba were studied regarding their reaction against superoxide (O2-) and hydroperoxyl radicals (HO2) in dimethyl sulfoxide as an aprotic solvent. It was found that the ginkgolides B, C, J, M as well as bilobalide react with superoxide and its protonated form as demonstrated by EPR and UV/VIS spectroscopy. The initial reaction rate with these oxygen-derived radicals is in the order of 100 M-1/s and below. Ginkgolide A does not react with superoxide under these conditions. From these findings it can be suggested that the superoxide scavenging effect of the ginkgolides B, C, J, M and bilobalide contributes to the antioxidant properties of G. biloba. PMID- 9413546 TI - Discrimination of carboxylesterases of chicken neural tissue by inhibition with a neuropathic, non-neuropathic organophosphorus compounds and neuropathy promoter. AB - Carboxylesterases are enzymes present in neural and other tissues that are sensitive to organophosphorus compounds. The esterase activity in particulate forms, resistant to paraoxon and sensitive to mipafox have been implicated in the initiation of organophosphorus-induced delayed polyneuropathy (OPIDP) and is called neuropathy target esterase (P-NTE). Certain esterases inhibitors such as phenylmethylsulfonyl fluoride (PMSF), can also irreversibly inhibit P-NTE and by this mechanism PMSF 'protects' from further effect of neuropathic OPs. However, if PMSF is dosed after a low non-neuropathic dose of a neuropathic OP, its neurotoxicity is 'promoted', causing severe neuropathy. The molecular target of promotion has not yet been identified and it has been shown that it is unlikely to be the P-NTE. In order to discriminate the different esterases, we used non neuropathic (paraoxon), and neuropathic organophosphorus compounds (mipafox, DFP) and a neuropathy promoter (PMSF). They were used alone or in concurrent inhibition to study particulate and soluble fractions of brain, spinal cord and sciatic nerve of chicken. From the experimental data, a matrix was constructed and equations deduced to estimate the proportions of the different potential activity fractions that can be discriminated by their sensitivity to the tested inhibitors. It was deduced that only combinations of up to three inhibitors can be used for the analysis with consistent results. In all tissues, inside the paraoxon sensitive activity, most of the activity was sensitive either to mipafox, to PMSF or both. In all fractions, except brain soluble fractions, within the paraoxon resistant activity, a mipafox sensitive component was detected that is operationally considered NTE (P-NTE and S-NTE in particulate and soluble fractions, respectively). Most of this activity was also sensitive to PMSF, and this should be considered the target of organophosphorus inducing neuropathy and of PMSF protective effect. Either in brain and spinal cord, a significant amount of the activity resistant to 40 microM paraoxon and 250 microM mipafox (usually called 'C' activity) is sensitive to PMSF. It could be a good candidate to contain the target of the promotion effect of PMSF as well as the S NTE activity that is also PMSF sensitive. PMID- 9413547 TI - Inhibition of thymidylate synthase and cell growth by the phenanthroindolizidine alkaloids pergularinine and tylophorinidine. AB - Biological activity of the phenanthroindolizidine alkaloids pergularinine (PGL) and tylophorinidine (TPD) isolated from the Indian medicinal herb Pergularia pallida has been evaluated and assessed for the first time employing thymidylate synthase (TS) (5,10-CH2H4 PteGlu: dUMP-C-methyltransferase, EC 2.1.1.45), a key target enzyme in cancer chemotherapy. TS used in the present investigations was purified from Lactobacillus leichmannii. Toxicity studies showed that PGL and TPD were potently toxic and inhibited growth of L.leichmannii cells. Both PGL and TPD significantly inhibited TS activity (IC50 = 40 and 45 microM, respectively). PGL concentrations > 80 microM and TPD concentrations > 90 microM resulted in a complete loss of the TS activity, thus suggesting that both these phenanthroindolizidine alkaloids are promising potential antitumor agents. Our results show that the alkaloid-binding to TS is irreversibly tight through a probable covalent linkage. Inhibition kinetics reveal that the enzyme has Ki values of 10 x 10(-6) and 9 x 10(-6) M for PGL and TPD, respectively and that the inhibition in both the cases is a simple linear 'noncompetitive' type. PMID- 9413548 TI - Dissimilarity in aflatoxin dose-response relationships between DNA adduct formation and development of preneoplastic foci in rat liver. AB - Earlier work in this laboratory and that carried out by others demonstrated that after a single dose of aflatoxin B1 (AFB) the resulting liver AFB-DNA adduct levels were directly proportional to dose. Earlier work also showed that after ten daily doses the AFB dose-response relationship with gamma-glutamyl transpeptidase (GGT) positive preneoplastic foci measured at 3 months was sublinear, with a threshold at a dose of about 150 micrograms/kg body weight/day. The objective of this study is to determine the factors influencing the shift in AFB dose-response between AFB-DNA adducts and GGT foci. Male Fisher 344 weanling rats were orally administered one or ten doses of AFB ranging from 50 to 350 micrograms/kg body weight/day. The animals were killed 2 or 24 h after the first AFB dose, or after the tenth AFB dose. The first and tenth doses were tritiated in these animals and 3H-AFB-guanine adducts isolated from liver DNA were measured by HPLC. Another group was killed 3 months after receiving ten doses in order to measure GGT foci development. AFB-guanine adduct levels were directly proportional to dose after the first dose, but after the tenth dose were much lower in the 200-350 micrograms/kg groups than after a single dose. The GGT foci response confirmed earlier work concerning a sublinear response. Among the individual animals in the 200-350 micrograms/kg groups there was a positive relationship, after controlling for dose, between GGT foci development and weight gained both during dosing (P = 0.018) and also to a lesser extent during the early promotional period (P = 0.066). Enzyme activity levels of GGT in liver homogenates were higher in the highest dose groups and reflected biliary proliferation rather than histological GGT stained foci. Urinary levels of AFB metabolites changed proportions in the high dosage multiply dosed animals reflecting alteration in AFB metabolism or excretion. The differences between the linear adduct and the sublinear foci dose-response curves may be the result of non-adduct effects of higher multiple AFB doses on foci formation including acute cytotoxicity, altered AFB metabolism and disposition, enhanced weight gains, or shortened foci latency but not through enhanced guanine adduct levels. Other studies that showed a linear relationship between AFB dose and liver tumor development used continuous feeding of maximal doses an order of magnitude less than the lowest dose in this study and thus avoided acutely toxic effects. We hypothesize that liver tumor development may mirror foci response in a 10-dose AFB regimen with doses above 100 micrograms/kg due to acute toxicity effects. PMID- 9413549 TI - Synthesis, structure and function of poly-alpha-amino acids--the simplest of protein models. AB - During the 1950s, linear and multichain poly-alpha-amino acids were synthesized by polymerization of the corresponding N-carboxyamino acid anhydrides in solution in the presence of suitable catalysts. The resulting homo- and heteropolymers have since been widely employed as simple protein models. Under appropriate conditions, poly-alpha-amino acids, in the solid state and in solution, were found to acquire conformations of an alpha-helix and of beta-parallel and antiparallel pleated sheets, or to exist as random coils. Their use in experimental and theoretical investigations of helix-coil transitions helped to shed new light on the mechanisms involved in protein denaturation. Conformational fluctuations of peptides in solution were analysed theoretically and studied experimentally by nonradiative energy-transfer techniques. Poly-alpha-amino acids played an important role in the deciphering of the genetic code. In addition, analysis of the antigenicity of poly-alpha-amino acids led to the elucidation of the factors determining the antigenicity of proteins and peptides. The synthetic procedures developed made possible the preparation of immobilized enzymes which were shown to be of considerable use as heterogeneous biocatalysts in the chemical and pharmaceutical industry. Interest in the biological and physicochemical characteristics of poly-alpha-amino acids was recently renewed because of the reported novel findings that some copolymers of amino acids are effective as drugs in multiple sclerosis, and that glutamine repeats and reiteration of other amino acids occur in inherited neurodegenerative diseases. PMID- 9413550 TI - Protein stability: still an unsolved problem. AB - This brief review suggests that molecular packing, the efficient filling of space, may be the most generally applicable factor that leads to the unique structures of most globular proteins. While simple in concept, the details of packing can lead to very subtle effects. The mechanical properties of a protein, dynamics and deformations under stress, tend to be asymmetric. In terms of structural alterations and thermostability, responses to genetic mutations are context dependent and remain difficult to predict with any confidence. Through small shifts proteins can frequently accommodate major changes in composition of the core region without substantial alteration in the basic chain conformation. Extending a jigsaw puzzle analogy, all of the pieces (side chains) are convex, varying flexible, and cannot be packed together without leaving cavities. Although large cavities do occasionally occur, a relatively even distribution of empty space is more common, and the overall packing does seem to specify the unique native structure. While it might appear that the translation machinery of the cell could have been designed with any set of alpha amino acids, the packing requirements, while strong, must be flexible enough to permit nondestructive single site mutations. This flexibility, combined with the need to produce a unique structure, may limit the average number of allowed side chain rotamers per residue. This in turn will reduce the allowable asymmetry of the side chains in order to maintain the largest number of structural motifs. It may be hard to improve on current set of amino acids. PMID- 9413551 TI - Organogenesis and angiogenin. AB - Our search for an angiogenesis-inducing factor in culture medium conditioned by human colon adenocarcinoma cells (HT-29) was inspired by the 'organizer' hypothesis originally postulated by Spemann. It led us to the isolation of angiogenin, a 14 kD protein homologous to pancreatic ribonuclease and one of the most potent stimulators of blood vessel formation known. This review summarizes the properties of angiogenin, its enzymatic and three-dimensional relationship to ribonuclease A (RNase A), those aspects of its structure that are critical for its biological function, and the therapeutic potential of angiogenin inhibition. Despite having the same arrangement of catalytic residues as RNase A, angiogenin has very low enzymatic activity. It lacks one of the four disulphide loops of RNase A; instead, the corresponding residues form part of a cell binding region. Both the catalytic activity and cell binding site are essential for angiogenesis. Angiogenin binds to cell-surface actin in confluent endothelial cells and to an as yet uncharacterized receptor on proliferating cells. Internalization and translocation to the nucleolus are also required for activity. Inhibitors of angiogenin can block angiogenesis in vitro and prevent tumour growth in vivo. Thus, a noncytotoxic neutralizing monoclonal antibody prevents the establishment of HT-29 human tumour xenografts in up to 65% of treated athymic mice. In those tumours that develop, the number of vascular elements is reduced. Actin also prevents the establishment of tumours while exhibiting no toxic effects at daily doses > 50 times the molar amount of circulating mouse angiogenin. These antagonists also inhibit the appearance of tumours derived from two other human tumour cell lines. Inhibition of the action of angiogenin may prove to be an effective therapeutic approach for the treatment of malignant disease. PMID- 9413552 TI - Psychrophilic enzymes: molecular basis of cold adaptation. AB - Psychrophilic organisms have successfully colonized polar and alpine regions and are able to grow efficiently at sub-zero temperatures. At the enzymatic level, such organisms have to cope with the reduction of chemical reaction rates induced by low temperatures in order to maintain adequate metabolic fluxes. Thermal compensation in cold-adapted enzymes is reached through improved turnover number and catalytic efficiency. This optimization of the catalytic parameters can originate from a highly flexible structure which provides enhanced abilities to undergo conformational changes during catalysis. Thermal instability of cold adapted enzymes is therefore regarded as a consequence of their conformational flexibility. A survey of the psychrophilic enzymes studied so far reveals only minor alterations of the primary structure when compared to mesophilic or thermophilic homologues. However, all known structural factors and weak interactions involved in protein stability are either reduced in number or modified in order to increase their flexibility. PMID- 9413553 TI - Endogenous trypsin receptors in Xenopus oocytes: linkage to internal calcium stores. AB - The effects of the protease trypsin, externally applied to full-grown oocytes of Xenopus laevis, were studied using electrophysiology and fluorometry. The following results were obtained: trypsin in concentrations of 0.1 microgram/ml to 1 mg/ml liberated Ca2+ from internal stores and evoked large transient currents of up to 5 microA in bath solutions containing 1 mM or no Ca2+. The response desensitized for 50 minutes and recovered at longer times. Transient currents could also be elicited by tryptic impurities in commercially available collagenase used for defolliculation of oocytes. Application of chymotrypsin (0.01 or 1 mg/ml) or of thrombin (3.4 ng/ml or 0.34 mg/ml) neither evoked currents nor desensitized trypsin responses. Incubation with 1 microgram/ml Pertussis toxin for 20 to 25 hours prevented the Ca2+ release from internal stores and the activation of transient currents by trypsin. We propose that endogenous receptors in the oolemma, specific for trypsin, are linked to internal Ca2+ stores via Pertussis toxin-sensitive G proteins. Thus, receptor activation by external trypsin raises internal Ca2+ and thereby opens Ca(2+)-activated Cl channels in the oolemma. PMID- 9413554 TI - Stability of solubilized benzodiazepine receptors. AB - According to the observations of other researchers, benzodiazepine receptors solubilized with sodium deoxycholate are unstable, but stability can be improved by exchanging deoxycholate for Triton X-100. In our experiments we conclude that the choice of detergent is not the restrictive factor for the stability of the solubilized receptors, but the storage conditions are. Solubilized receptors, either stored in sodium deoxycholate or in Triton X-100, were stable for at least 2 months when stored at -80 degrees C, but both preparations were strongly unstable when stored at -20 degrees C. Alternatively, sodium deoxycholate solubilized receptors may be lyophilized and then stored at -20 degrees C. PMID- 9413555 TI - Purification of bovine alpha-lactalbumin by immobilized metal ion affinity chromatography. AB - The milk protein alpha-lactalbumin was isolated from bovine whey protein concentrate solution by immobilized metal ion affinity chromatography (IMAC) using Cu(II)-Chelating Sepharose Fast Flow. Stepwise pH (5.5-3.8) changes in sodium acetate buffer were used to elute the protein selectively, at which time it was concentrated and reapplied to an uncharged Chelating Sepharose Fast Flow column to remove the contaminating Cu(II) ions. A purity of 90% and recovery of 80% was achieved. The described method appears to be suitable for isolation of alpha-lactalbumin in a form adequate for milk formula engineering. PMID- 9413556 TI - Purification of human alpha 2-antiplasmin with chicken IgY specific to its carboxy-terminal peptide. AB - alpha 2-antiplasmin, a plasma glycoprotein of the serpin superfamily, is the primary physiological inhibitor of plasmin, the key enzyme in fibrin degradation. Previous purification methods utilize lengthy multistep protocols with low yields or use monoclonal antibodies that are expensive or difficult to make. With a relatively small investment, a chicken was immunized with keyhole limpet hemocyanin-conjugated to alpha 2-antiplasmin C-terminal 26 residue synthetic peptide and the peptide-specific antibody (IgY) was isolated from the egg yolks of hens using the peptide affinity column. Based on the interaction between this IgY and alpha 2-antiplasmin, pure alpha 2-antiplasmin was isolated from human plasma in two steps: (a) citrated plasma was precipitated with 15% PEG-8000 to remove the bulk of plasma proteins while retaining the majority of alpha 2 antiplasmin activity; and (b) the alpha 2-antiplasmin was affinity-purified from the supernatant using the IgY column. Yields were typically 48% and the purity and authenticity of the alpha 2-antiplasmin were verified by gel electrophoresis, Western Blot analysis, N-terminal sequence, and amino acid analysis. PMID- 9413557 TI - Purification and characterization of protein D/E, a putative sperm-binding protein involved in fertilization. AB - We describe a method for the efficient purification of a 32 Kd glycoprotein from rat epididymal tissue. The glycoprotein was purified by gel filtration, ion exchange, affinity, and reverse phase high pressure liquid chromatography. The highly purified glycoprotein was radiolabeled with an iodinatable, cleavable, photoreactive cross-linking agent, 1-[N-(2-hydrox-5-azidobenzoyl)-2-aminoethyl]-4 (-hydroxysuccini mid yl)-succinate (HAHS). The soluble radiolabeled glycoprotein was bound to washed epididymal spermatozoa in a time-dependent, saturable, and reversible manner. Scatchard analysis demonstrates that there are approximately 3,403 binding sites/spermatozoon. The binding efficiency (Kd) for spermatozoa was approximately 2.0 x 10(-10) M. The function of this glycoprotein was verified by using an in vivo artificial insemination fertilization assay. The fertility rate for control spermatozoa was approximately 53%, but the rate for spermatozoa exposed to polyclonal anti-glycoprotein antibodies was only 5%. These data suggest that the binding of the glycoprotein to the surface of rat spermatozoa is mediated by a receptor-type mechanism and is involved in the fertilization process. PMID- 9413558 TI - Purification and characterization of a 31-kilodalton iron-regulated periplasmic protein from Pasteurella haemolytica A1. AB - A prominent iron-regulated periplasmic protein was purified from Pasteurella haemolytica grown in an iron-deficient chemically defined medium. The protein was purified by anion exchange chromatography and appeared as a single band by SDS PAGE with a molecular weight of 32,000. A yield of five mg was obtained from 91 mg of protein extract. The iron-regulated protein existed as a monomer in the native state with an average molecular weight of 29,877 as determined by analytical ultracentrifugation. The protein had a molecular weight of 30,880 as determined by matrix-assisted laser desorption mass spectrometry, hence the protein is referred to as the 31 kDa protein. Isoelectric focusing showed four bands with pIs of 7.15, 6.8, 6.6, and 5.9. The secondary structure of the protein was determined by circular dichroism and contained 16% alpha-helical structure. The N-terminal sequence, EPFKVVTTFTVIQDIAQNVAGDKAT, showed a 95% identity with the 31 kDa iron-binding protein from Haemophilus influenzae. Isolation and characterization of iron-regulated proteins are of particular interest because of their potential roles in iron assimilation and microbial virulence. PMID- 9413559 TI - Carbonic anhydrase from Camelia sinensis (tea) leaves. AB - Carbonic anhydrase (CA) (carbonate hydrolyase; E,C,4.2.1.1) from leaves of mature Camelia sinensis was purified and characterized. The purification level was 53 fold. The optimum temperature for maximal enzyme activity is 50 degrees C. The optimum pH was 6.8 and this pH varied between 6.5 and 7.5. Each enzyme molecule is a hexamer with an M(r) of 169,000 with subunits of M(r) = 28,000. PMID- 9413560 TI - A different structural feature for carbonic anhydrases in human erythrocytes. AB - This study presents a different structural feature for carbonic anhydrase in human erythrocytes. Carbonic anhydrase isozymes (CA-I and CA-II) were purified from an erythrocyte pool of 20 healthy subjects. For purification, Sepharose-4B-L tyrosine-sulfanilamide affinity column was used. Resnets from 3-10% discontinuous SDS-polyacrylamide gel electrophoresis (SDS-PAGE) showed a single band for CA-I and two distinct bands for CA-II. The molecular weights of the two bands were similar. One peak for CA-I and two peaks for CA-II were obtained in gel filtration. The enzymatic activities of the bands in question were also of different value. Native electrophoresis showed two bands for CA-I, and it showed three bands for CA-II. It can be concluded that CA-I is a polymer composed of a single promoter and CA-II has three different polymers composed of two distinct promoters, suggesting a new structural feature of human erythrocyte carbonic anhydrase isozymes. PMID- 9413561 TI - Isolation of myelin basic protein from whole tissue extracts by selective pH dependent solubilization. AB - A previous study of the selective solubility of myelin basic protein (MBP) of tissue extracts at pH 9.0 has raised issues of its quantitative recovery, and the differential solubility of its charge isomers. The pH-dependent solubility of proteins of acid extracts of delipidated tissue of bovine spinal cord was therefore reexamined. MBP of whole extracts was completely soluble up to pH 8.0 only, and less so by 25% at pH 9.0, and 43% at pH 10.0. The proteins other than MBP were virtually insoluble between pH 5.0 to 6.0, and 9.0 to 10.0. The solubility of the main charge isomers I to III of MBP of 18.5 kDa was found not to be affected by pH. Either pH 5.0 or 9.0 is therefore suitable for the selective isolation of MBP from whole tissue extracts, only pH 5.0 providing for the complete recovery of MBP. The pH-dependent solution behaviour was also examined following the separation of proteins of whole extracts by anion exchange chromatography at pH 10.4. Purified MBP and several related minor cationic components of lower molecular weight were soluble throughout. In contrast, the anionic proteins were only partly soluble between pH 4.0 to 10.0, i.e. by 4 to 20%. The results are consistent with specific protein-protein interactions of the proteins of whole extracts, either enhancing the solubility of non-MBP proteins, e.g. at pH 7.0, or impairing that of MBP between pH 8.0 to 10.0. PMID- 9413562 TI - Genes controlling neural fate and differentiation. PMID- 9413563 TI - Neuronal plasticity in development: lessons from ethanol neurotoxicity during embryogenesis. PMID- 9413565 TI - Neuron-glia cross talk in rat striatum after transient forebrain ischemia. PMID- 9413564 TI - Plasticity in astrocytic phenotypes. A role for protein kinase C, tyrosine kinases, and cytoskeleton signaling. PMID- 9413566 TI - Regulation of oligodendrocyte differentiation by fibroblast growth factors. PMID- 9413567 TI - Features and functions of human microglia cells. PMID- 9413568 TI - Migration and proliferation of mononuclear phagocytes in the central nervous system. PMID- 9413569 TI - K(+)-channels and cytokines as markers for microglial activation. PMID- 9413571 TI - Regulation by neurotransmitters of glial energy metabolism. PMID- 9413570 TI - Anatomical and functional characteristics of transplanted monoaminergic neurons in paraplegic rats. PMID- 9413572 TI - Dendritic development of visual callosal neurons. PMID- 9413573 TI - Neuronal and non-neuronal plasticity in the rat following myenteric denervation. PMID- 9413574 TI - Nerve growth factor and oxidative stress in the nervous system. PMID- 9413575 TI - Role of astrocytes in glutamate homeostasis. Implications for excitotoxicity. PMID- 9413576 TI - Nerve cell death induced by Ca2+ ionophores in dissociated hippocampal cultures. Protective action of the NMDA antagonist MK-801. PMID- 9413578 TI - Alzheimer disease, from molecular biology to therapy. PMID- 9413577 TI - Is increased neurotoxicity a burden of the ageing brain? PMID- 9413579 TI - Steroid and protein regulators of glial cell proliferation. AB - Any variation in CNP and GS activity in vitro under the effect of growth factors with different mechanisms of action, may indicate a general process of differentiation in the culture. An increase in the GS activity in early passage glioma may be interpreted as an induction of the differentiating phenotype involving cell transformation into a more astrocytic-like culture. Hence, under the above experimental conditions, dexamethasone and insulin are the two factors with the most potent differentiating effect on rat C6 glioma cell line. Comparative studies with growth regulators using different receptor pathways and mechanisms of action may add valuable data to the current knowledge in the field. The data presented in this chapter show the plasticity of C6 glioma to the differentiating effect of various growth factors, especially insulin and dexamethasone, and confirms C6 glioma cell line as a useful model for studies on glial cell properties and proliferation in vitro. PMID- 9413580 TI - Estrogens influence growth, maturation, and amyloid beta-peptide production in neuroblastoma cells and in a beta-APP transfected kidney 293 cell line. AB - During development in vivo and in vitro, estrogens: a) increase brain excitability, particularly in limbic structures; b) are responsible for the maturation and cyclicity of limbic-hypothalamic interrelations; c) enhance myelinogenesis; and d) may act with NGF to stimulate neurite formation. In senescence, estrogen administration would improve memory in postmenopausal women. The absence or low levels of estrogens after menopause would increase prevalence of Alzheimer's dementia (AD) more in women than men, irrespective of age or ethnicity. In the present study, addition of 17-beta estradiol to cultured human neuroblastoma cells affected growth slightly, but stimulated cell maturation as shown by increased tyrosine hydroxylase activity. The extracellular deposition in brain tissue and around blood vessels of the amyloid beta-peptide (A beta), a 4.3 kD fragment of the larger integral membrane protein, beta-amyloid precursor protein (beta-APP), is considered an important characteristic of AD. We investigated whether 17-beta estradiol may influence the formation of the A beta (thus the abnormal accumulation of amyloid proteins) in neuroblastoma cells and in a beta-APP transfected human kidney 293 cell line. Two doses of 17 beta estradiol were added to the cultures of both cell lines. Cells were grown until confluence, metabolically labeled with 35S-methionine, immunoprecipitated with the rabbit antiserum R1282, gel electrophoresed and autoradiographed in order to compare levels of A beta under the different estradiol concentrations. While in neuroblastoma cells, levels of A beta were only slightly reduced after estradiol and a dose-effect relationship with the hormone could not be demonstrated, in the 293 cells, A beta band intensity decreased as concentration of estradiol increased. These data support the role of estrogen in normal and abnormal brain metabolism and suggest potential hormonal interventions which may reduce or prevent the formation of amyloid deposits occur in AD. PMID- 9413581 TI - Role of testosterone in the activation of sexual behavior and neuronal circuitries in the senescent brain. PMID- 9413582 TI - Endogenous opioids and prenatal determinants of neuroplasticity. PMID- 9413583 TI - Brain plasticity and the neural cell adhesion molecule (NCAM). PMID- 9413584 TI - Pain. PMID- 9413585 TI - Femoral sulcus angle measurements. PMID- 9413586 TI - The value of postmortem retrievals in the analysis of long-term, well-functioning cemented femoral stems. AB - Multiple sections from long-term, low-friction bilateral Charnley prostheses harvested from two patients who died 20 and 22 years after the primary implantation were studied. All sections showed generally sound mechanical integrity at both the cement-bone and cement-metal interfaces, in spite of localized evidence of cement fracture and debonding. The host tissues showed no clear evidence of an adverse biologic response toward the implant. These findings aided in the delineation of the sequence of biologic and mechanical events that lead to prosthesis failure. They suggest that if the apparently unavoidable mechanical deterioration of cemented implants can be limited by good fixation between the host and prosthesis, then disastrous biologic events, such as osteolysis, may be avoided, and long-term function, preserved. PMID- 9413588 TI - Joint moments in minor limb length discrepancy: a pilot study. AB - A biomechanical analysis was performed to determine if a minor limb length discrepancy alters lower extremity joint mechanics significantly and in a manner that could contribute to the development of joint abnormalities. Ten healthy subjects with equal limb lengths were recruited. Gait analysis was performed for both left and right sides to determine the maximum moments at the hip, knee, and ankle joints. A minor limb length discrepancy was simulated by adding a shoe lift of 1.25 cm to the left leg. After a period of acclimation, the gait was reanalyzed. Differences for maximum joint moments at the hip, knee, and ankle before and after simulation were nonsignificant. An additional 10 healthy, asymptomatic patients with actual limb length discrepancies ranging from 1 cm to 2 cm were also recruited. Gait analysis for maximum joint moments before and after correction of the limb length discrepancy was performed. Side-to-side differences in joint moments before correction were nonsignificant. After correction of the limb length discrepancy, side-to-side joint moment differences were significantly increased (P = 0.02) and may suggest acute overcompensation to the presence of the corrective shoe lift. Consequently, this study did not find an association between minor limb length discrepancies and predictable changes in lower extremity joint kinetics that might potentially lead to joint abnormalities. PMID- 9413587 TI - A second-generation cementless hip prosthesis: improved results over the first generation prosthesis. AB - Forty-two patients who had a Porous Coated Anatomic (PCA) "E" series, second generation, cementless hip arthroplasty (Howmedica, Rutherford, NJ) were compared with 42 patients who had a first-generation PCA prosthesis. Patients were directly matched for age, sex, diagnosis, weight, Charnley functional status, and duration of follow-up. All of the operations were done by the same two surgeons, who used the same operative approach and the same postoperative rehabilitation plan. All of the patients were followed up for at least 5 years (range, 60 to 76 months). In the "E" series group, there were 41 of 42 (98%) good and excellent clinical results with a mean Harris hip score of 94 points (range, 46 to 100 points); the first-generation group had 34 of 42 (81%) good and excellent clinical results and a mean Harris score of 81 points (range, 42 to 100 points) (P = 0.001). There was one acetabular component revision in the "E" series group (2%), which can be compared with eight revisions (19%) in the first-generation group (P = 0.012). The incidence of femoral radiolucencies was 19% (eight hips) for the "E" series group compared with 50% (21 hips) in the first-generation group (P = 0.009). The radiolucencies in the "E" series group were small, nonprogressive, and confined typically to zone I. We believe that the improvements in design of the "E" series component may account for these differences. PMID- 9413589 TI - Concurrent rupture of the Achilles and plantaris tendons. PMID- 9413590 TI - Snowblower injuries to the hand. AB - We retrospectively reviewed the records of 62 patients who sustained serious hand injuries caused by snowblowers between 1981 and 1990. Frequency of injuries to digits tended to correlate with length (i.e., middle, index, ring, or small finger or thumb). Damage to tendons did not seem to follow any particular pattern. The majority of victims sustained multiple digital involvement. Complete versus partial amputation followed the same length distribution as did injured digits. Most of the injuries occurred to the dominant hand. When patients were further questioned regarding the circumstances and events leading to their injury, a recurring pattern was found. Most patients described a wet, heavy snow having recently fallen. The majority of the patients who were injured by placing their hands into the exit chute admitted that they were aware the machine was running, but thought that they had a greater clearance to the rotating impeller blade. PMID- 9413591 TI - Hyphenated history: the Phelps-Baker test. AB - The language of orthopedics is filled with eponyms. Orthopedic surgeons speak cryptically to one another using code words and "orthopedic pig-Latin." Certain hyphenated eponyms are particularly interesting, because they represent people who came to be partners in orthopedic history. The derivation of the Phelps-Baker test, an important component of the hip examination of children who suffer from cerebral palsy, named in honor of Winthrop Morgan Phelps and Lenox D. Baker, is described in this report. PMID- 9413592 TI - Silica induces changes in cytosolic free calcium, cytosolic pH, and plasma membrane potential in bovine alveolar macrophages. AB - The mineral-dust induced activation of pulmonary phagocytes is thought to be involved in the induction of severe lung diseases. The activation of bovine alveolar macrophages (BAM) by silica was investigated by flow cytometry. Short term incubation (< 10 min) of BAM with silica gel and quartz dust particles induced increases in the cytosolic free calcium concentration ([Ca2+]i), decreases in intracellular pH (pHi), and increases in plasma membrane potential (PMP). The extent of these changes was concentration dependent, related to the type of dust and was due to Ca2+ influx from the extracellular medium. An increase in [Ca2+]i was inhibited, when extracellular Ca2+ was removed. Furthermore the calcium signal was quenched by Mn2+ and diminished by the calcium channel blocker verapamil. The protein kinase C specific inhibitor bisindolylmaleimide II (GF 109,203 X) did not inhibit the silica-induced [Ca2+]i rise. In contrast, silica-induced cytosolic acidification and depolarization were inhibited by GF 109,203 X but not by removal of extracellular calcium. Addition of TiO2 particles or heavy metal-containing dusts had no effect on any of the three parameters. Our data suggest the existence of silica-activated transmembrane ion exchange mechanisms in BAM, which might be involved in the specific cytotoxicity of silica by Ca(2+)-dependent and independent pathways. PMID- 9413593 TI - Parameters derived from integrated nuclear fluorescence, syntactic structure analysis, and vascularization in human lung carcinomas. AB - Combined measurements of integrated nuclear fluorescence (INF) and vascularization were performed on surgical specimens of human lung carcinomas. Histological slides of formalin-fixed, paraffin-embedded tissue samples were treated with Texas Red-labeled antibody to factor VIII and the fluorochrome DAPI. The resulting images were analyzed with an epi-illumination fluorescence microscope and two different filter blocks. The first image displayed the vessels, and the second the DAPI-stained nuclei of surrounding cells. The extent of vascularization was assessed by calculating the volume fraction (Vv), the surface fraction (Sv), the area, and the minimum diameter of the vessels. The INF was measured in tumour cells and lymphocytes, and was grouped according to the distance from the nearest vascular boundary into the intervals of 0-20, 21-40, 41 60, 61-80, and > 80 mu. The numerical densities (Nv) as well as the percentages of S-phase-related tumour cell fraction (SPRF) and of tumour cells with an INF > 5C were computed. A minimum of 50 vessels and 300 tumour cells were examined. The material included 100 cases with primary lung carcinoma (39 epidermoid carcinomas, 39 adenocarcinomas, 13 large cell carcinomas, three small cell anaplastic carcinomas, and 6 carcinoid tumours). On the average, the volume density of the stroma amounts to 16.7%, and that of the vessels (Vv) to 12.8%. The minimum diameter of the intratumoral vessels is 13 mu and the measured circumference 138 mu. The numerical densities of tumour cells (lymphocytes) decrease with increasing distance from the vascular boundary from 6.3 (1.7) to 1.0 (0.1). A reduction is also seen in the percentage of the SPRF from 10.7 to 8.1%. The percentage of tumour cells with an INF > 5C, however, is positively correlated to the distance from the vascular surfaces from 34.2 to 38.2%. The measurements reveal that tumour cells are densely positioned and have an increased proportion of proliferation in the populations close to perivascular spaces, whereas chromosome abnormalities are seen more frequently, when tumour cells are located at a distance > 20 mu from the vascular surfaces. PMID- 9413594 TI - Correlation between p53 status, DNA ploidy, proliferation rate and nuclear morphology in breast cancer. An image cytometric study. AB - The study was designed to detect differences in the nuclear morphology of tumours and tumour cell populations with different p53 expression in correlation with DNA ploidy and proliferation rate. The paraffin sections from routinely processed samples of 88 breast cancers were immunostained with the monoclonal p53-antibody DO-1. After localization and evaluation with a scoring system the sections were destained and stained by the Feulgen method. The nuclei were relocated automatically and measured by means of the image cytometry workstation. Significant differences between the tumours and tumour cell populations with different p53 expression were found in the euploid tumours as well as in the aneuploid tumours and in the breast cancers with a high proliferation rate. The breast cancers with a low immunoreactive score (IRS 1-4) differ from the negative cancers as well as from the cancers with a higher immunoreactive score (IRS 5 12). Evaluating the nuclear populations of the p53 positive cancers, there were differences in the features of the chromatin amount and distribution in the groups of the euploid breast cancers and in cancer with a high proliferation rate. In contrast, the nuclear populations of the aneuploid cancers did not show any differences in their nuclear morphology. The results showed the different impacts of the p53 expression, DNA ploidy and the proliferation rate on the nuclear morphology in breast cancer. PMID- 9413595 TI - Aneuploidy mechanisms in human colorectal preneoplastic lesions and Barrett's esophagus. Is there a role for K-ras and p53 mutations? AB - The link of aneuploidy and heteroploidy in human solid tumours with early genetic events is poorly understood. The study of human preneoplastic precursor lesions, i.e., colorectal adenomas, chronic ulcerative colitis lesions, and Barrett's esophagus, as considered in this review, appears particularly useful to achieve this aim. Literature data examined here on aneuploidy were obtained by image and flow cytometry, classical cytogenetics, and in situ hybridization based cytogenetics. It appears that aneuploidy is linked with specific gene mutations, i.e., of the tumour suppressor gene p53 in chronic ulcerative colitis and in Barrett's esophagus, and of the protooncogene K-ras in colorectal adenomas. These data and data from experiments using in vitro and mouse models, suggest that chromosome instability, tetraploidization, and asymmetrical chromosome segregation during cell division are the result of deregulated cell cycle genes with multiple functions that normally exert active checks on the cell cycle processes including apoptosis and chromosome stability. PMID- 9413596 TI - Quantitative comparison of immunohistochemical staining intensity in tissues fixed in formalin and Histochoice. AB - Formaldehyde fixatives have traditionally been used to preserve tissues as they impart excellent morphological preservation. Formaldehyde fixes tissue by cross linking, a process which can reduce the antigenicity of tissue and weakens the intensity of immunohistochemical stains. Preliminary studies have shown that Histochoice tissue fixative offers equal or greater staining intensity than neutral buffered formalin (NBF). This study compares these fixatives quantitatively and presents the results in unambiguous statistical terms. Tissue samples were collected, bisected, and fixed in NBF or Histochoice. The sections were stained with a total of 21 antibodies, and color images were collected. The hue, saturation, and value were determined for each positive pixel and an ANOVA performed. Small differences in hue were noted in 8 of 21 cases. Histochoice fixed tissue gave a greater mean saturation than NBF with 57.1% of the antibodies tested. No significant difference in the saturation was detected in 28.6% of the cases; NBF gave higher mean saturation levels with only 14.3% of the antibodies. Histochoice-fixed tissue was found to give lower values in 66.7% of cases than those prepared with NBF, indicating darker staining. These results show that Histochoice produces staining intensity that is comparable, and in many cases superior, to formalin. PMID- 9413597 TI - Resolve to join the quest for valid patient outcome measures. PMID- 9413598 TI - A collaborative approach to minimally invasive direct coronary artery bypass. AB - Minimally invasive direct coronary artery bypass (MIDCAB) graft is a surgical technique that is becoming more widely accepted. Through a collaborative effort- interhospital and intrahospital--surgical team members at two hospitals in Kentucky made significant improvements on the MIDCAB procedure that positively influenced patient outcomes (e.g., less time in intensive care, shorter hospital stays, fewer complication, cost savings to the patients and institutions). This article reviews those collaborative efforts and outcomes. PMID- 9413599 TI - Minimally invasive treatment for coronary artery disease. AB - Coronary artery disease is the leading cause of morbidity and mortality in the United States. One advancement in the treatment of this disease is the minimally invasive direct coronary artery bypass (MIDCAB) procedure, which is an alternative to the traditional open heart bypass procedure. The MIDCAB procedure is becoming a viable alternative to the traditional coronary artery bypass grafting procedure for a select group of patients. With further experience and follow-up, this procedure will offer lower hospital costs by decreasing lengths of stay and offering patients the optimal conduit for coronary artery bypass grafting without the complications of cardiopulmonary bypass. PMID- 9413600 TI - Minimally invasive direct coronary artery bypass surgery. AB - Traditional coronary artery bypass surgery involves a median sternotomy and the use of a heart-lung machine to stabilize the heart during suturing. Minimally invasive coronary artery surgery employs small incisions directly over the target vessels and avoids the use of a heart-lung machine, which can cause postoperative complications. The target coronary vessels are stabilized in alternative ways, potentially hazardous manipulation of the ascending aorta is avoided, and the subclavian and axillary arteries provide alternative inflow sources. Other new techniques used in minimally invasive procedures include a coronary artery cannula to avoid intraoperative ischemia and wound irrigation catheters to administer postoperative bupivacaine hydrochloride. Perioperative nurses need to become familiar with these new techniques to be able to plan and implement effective patient care. PMID- 9413601 TI - Improving glove barrier effectiveness. AB - Perioperative staff members depend on surgical gloves to prevent disease transmission between themselves and patients, but these gloves frequently fail during use. Three approaches can make surgical gloves more effective barriers: preventing glove failures, monitoring glove integrity, and improving glove quality. Failure prevention includes modifying surgical techniques, improving instruments and equipment, streamlining teamwork, selecting the most appropriate gloves, double gloving, and performing preventive glove changes. Glove integrity monitoring can be performed visually or by feel, by wearing glove pairs with color-puncture indicators, or by using electronic monitoring devices. Glove quality improvements must be accompanied by testing methods that reflect in-use conditions. A glove rating system that is based on in-use performance may enhance glove safety substantially. PMID- 9413602 TI - Building a successful risk-based competency assessment model. AB - The requirement to verify and ensure the competency of staff members to perform their assigned duties is here to stay. This article describes a model for decision making about competency assessment frequency. Implementation of the model should be accompanied by a systematic review of learning needs and performance improvement data. Education sessions designed to address identified learning needs or to support performance improvement activities should occur before or concurrent with competency assessment. The result is a cost-effective, efficient use of resources to accomplish the goal of ongoing assessment and improvement of staff competency. When staff member competency improves, the likelihood of a positive patient outcome increases. Our model provides a structured, defensible mechanism to link competency assessment with improvements in patient care quality. PMID- 9413603 TI - Is it only a tonsillectomy? PMID- 9413604 TI - Measurement reliability and validity. PMID- 9413605 TI - The Colorectal Cancer Recurrence Support (CARES) System. AB - Colorectal cancer has risen in incidence to become the second commonest form of cancer in Singapore. The primary treatment is surgery but up to 50% of patients still suffer from recurrence of the cancer after surgery. Early identification of recurrence will increase the effectiveness of therapy and the survival of patients. This paper describes the CARES (Cancer Recurrence Support) System, whose objective is to predict the recurrence of colorectal cancer, using Case based Reasoning (CBR), and supported by other techniques such as data mining and natural language processing. The CARES System employs CBR to compare and contrast between the new and past colorectal cancer patient cases, and makes inferences based on those comparisons to determine the high risk patient groups. The features and functionality of the system are described. PMID- 9413606 TI - Formalisation for decision support in anaesthesiology. AB - This paper reports on research for decision support for anaesthesiologists at the University Hospital in Groningen, the Netherlands. Based on CAROLA, an existing automated operation documentation system, we designed a support environment that will assist in real-time diagnosis. The core of the work presented here consists of a knowledge base (containing anaesthesiological knowledge) and a diagnosis system. The knowledge base is specified in the logic-based formal specification language AFSL. This leads to a powerful and precise treatment of knowledge structuring and data abstraction. PMID- 9413607 TI - Application of autonomous neural network systems to medical pattern classification tasks. AB - This paper presents a study of the application of autonomously learning multiple neural network systems to medical pattern classification tasks. In our earlier work, a hybrid neural network architecture has been developed for on-line learning and probability estimation tasks. The network has been shown to be capable of asymptotically achieving the Bayes optimal classification rates, on line, in a number of benchmark classification experiments. In the context of pattern classification, however, the concept of multiple classifier systems has been proposed to improve the performance of a single classifier. Thus, three decision combination algorithms have been implemented to produce a multiple neural network classifier system. Here the applicability of the system is assessed using patient records in two medical domains. The first task is the prognosis of patients admitted to coronary care units; whereas the second is the prediction of survival in trauma patients. The results are compared with those from logistic regression models, and implications of the system as a useful clinical diagnostic tool are discussed. PMID- 9413608 TI - Application of the fuzzy ART/MAP and MINMAX/MAP neural network models to radiographic image classification. AB - This paper concerns the classification analysis of exercise-induced lower leg pain by applying competitive neural network clustering and mapping techniques to type 1 and type 2 fuzzy descriptions of bone scan images of the tibia. The clusters are described and compared with each other and with the experts known classes that would be expected from medical findings. The discovered clusters provide training sets for supervised learning by an ARTMAP and similar neural network. These were used to classify the previously unclassified images and hence improve the classification process. The overall conclusion is that the use of the neural clustering methods has improved the classification process of the shin images despite the paucity of data and its inherent uncertainty. PMID- 9413609 TI - Optimization of the separation selectivity of PCBs in a hydroorganic reverse phase liquid chromatography in the presence of cetyltrimethylammonium bromide with fluorescence detection. AB - The retention of different PCBs of toxicological and environmental interest was studied in a hydro-alcoholic reverse-phase liquid chromatography in the presence of cetyltrimethylammonium bromide (CTAB). The influence of variables like type and percentage of alcohol, concentration of CTAB, temperature and mobile phase flow was determined. According to the aforementioned studies, the appropriate conditions for separation of PCBs in presence of CTAB in a hydro-alcoholic medium are: percentage of 1-Propanol 55% (v/v), concentration of CTAB 5.0 x 10(-3) M, temperature 50 +/- 1 degrees C and mobile phase flow 1 ml min-1. PMID- 9413611 TI - Identification and characterization of recombinant murine interleukin-6 with a C terminal pentapeptide extension using capillary reversed phase HPLC-MS and edman degradation. AB - We have identified a preparation of recombinant murine interleukin-6 (mIL-6) that, in addition to the anticipated product, also contained approximately equal amounts of mIL-6 with a C-terminal pentapeptide extension. The extension mutant was generated by readthrough of the stopcodon, and termination at a second in frame stopcodon 12 base pairs 3' in the expression vector. Aliquots of the preparation were subjected to proteolytic digestion with Asp-N and Lys-C endopeptidase. The resultant peptides were separated by reversed-phase capillary HPLC, and analysed using a combination of mass spectrometry and N-terminal sequence analysis. These data revealed a C-terminal pentapeptide (Gln-Gly-Ser-Val Asp) extension, with the authentic stopcodon being translated as glutamine. The extension mutant was isolated by reversed-phase HPLC and shown to have similar mitogenic activity to mIL-6 on murine hybridoma 7TD1 cells. PMID- 9413612 TI - Partitioning of flunitrazepam into model membranes studied by temperature controlled gel filtration chromatography. AB - The use of gel filtration chromatography with Sephadex as a separation medium was used in order to study flunitrazepam (FNTZ) partitioning into artificial model membranes consisting of dipalmitoyl-phosphatidylcholine (dpPC) vesicles, under controlled temperature conditions. In this system two phenomena are taking place simultaneously: the ligand-liposome interaction and the lipid self-aggregation to form the liposome. The liposome-FNTZ interaction was evidenced by the non enantiography of the first derivative of FNTZ elution peak in frontal chromatography through Sephadex G-75. On the other hand, the presence of FNTZ reduced liposomes mean size and increased their size dispersion as evidenced by molecular filtration through Sephadex G-200. The dpPC-buffer FNTZ partition coefficient determined in zonal chromatography through Sephadex G-10 increased about 33% when the temperature rose above the temperature of dpPC transition from the liquid crystalline to the fluid phase. Gel filtration chromatography seems a suitable technique to study lipid liposome-FNTZ interactions at a qualitative level. In addition, this technique has the advantage over other methods of giving the possibility of observing the mutual effects exerted between the drug and the self-aggregating structure. PMID- 9413610 TI - Determination of free amino acids in microalgae by high-performance liquid chromatography using pre-column fluorescence derivatization. AB - An analytical method for a rapid reverse-phase liquid chromatography of amino acids is presented. The total analysis time was 19 min. The average relative deviation over the measured peak area was lower than 10% and the fluorescent response was linear with concentration for OPA derivatives (r > 0.99) in the range 100-1000 nM. The proposed method was applied to the determination of free amino acids in two microalgae species (Chlorella pyrenoidosa and Chlamydomonas rinhardtii). The principal aminoacids found were alanine, arginine leucine, serine and glutamic acid. PMID- 9413613 TI - Effective one/two step purification of various materials by solid-phase extraction. AB - Simple one/two step purification procedures based on the solid-phase extraction technique were effectively exploited to clean up radiolabelled drugs represented by dihydrochloride of [6-3H]-stobadine and hydrochloride of [4-3H]-pentacaine, derivatization agents such as 4-nitrobenzoyl chloride or 3,5-dinitrobenzoyl chloride, as well as the aqueous phosphate or triethylamine acetate buffer solutions. PMID- 9413614 TI - Identification of metabolites of malathion in plant, water and soil by GC-MS. AB - Metabolism of malathion (O,O,-dimethyl S-[1,2-bis(ethoxy carbonyl) ethyl]phosphorodithioate) has been studied in plants, water and soil. A number of products identified by gas chromatography-mass spectrometry (GC-MS), have been formed by de-esterification, oxidation and hydrolysis of malathion. Hydrolysis of the P-S and C-S bonds occur in alkaline pH 8.0 and acidic pH 5.5, respectively. PMID- 9413615 TI - Stereoselective metabolic study of famprofazone. AB - Famprofazone (1) metabolites were studied in human urine after medication by 50 mg oral dose. The human urine was collected over 48 h from six volunteers at time intervals of 6, 12, 24 and 48 h. The amount of famprofazone metabolites were recovered from the urine samples by application of Extrelut extraction method. The resultant extracts were derivatized using N-methyl-N trimethylsilytrifluoroacetamide (MSTFA) for trimethylsilylation followed by N methyl-bis-trifluoroacetamide (MBTFA) for trifluoroacetylation. Methamphetamine (2) and 3-hydroxymethyl-propyphenazone (3), excreted in human urine, were identified as famprofazone metabolites by gas chromatography-mass spectrometry (GC-MS). The quantitative results revealed that the average amounts of 2 and 3, excreted in human urine were equal to 2.6 and 4 mg, respectively, through 48 h. However, 3 was analysed after enzymatic hydrolysis of the urine samples using beta-glucuronidase/arylsulphatase. The excreted methamphetamine enantiomers could be separated by application of indirect GC-technique using S-(-)-N trifluoroacetylprolyl chloride (TPC) as a chiral derivatizing agent. The average amount of (-)-methamphetamine isomer excreted in the urine was found to be three fold those of the (+)-isomer. PMID- 9413616 TI - HPLC/circular dichroism detection using a split-type flow cell for polarized photometric detector. AB - HPLC/circular dichroism detection using a conventional spectrophotometric detector has been established. A split-type flow cell developed for a polarized photometric detector, which is a non-modulated polarimeter, was inserted into this detector. When two rays of circularly polarized light with opposite rotational senses to each other were incident in both cells by combining a polarizer and a 1/4 wave retarder in front of the cell assembly, the output from this spectrophotometric detector became solely the signal based on circular dichroism. Selective detection of a chiral analyte, in which the ratio of the difference in the absorbance between two rays mentioned above and the absorbance by normal rays in over 1/1000, is feasible using the proposed instrument. PMID- 9413617 TI - Separation of porphyrins by capillary electrophoresis in fused-silica and ethylene vinyl acetate copolymer capillaries with visible absorbance detection. AB - A mixture of 5 porphyrins were separated by capillary electrophoresis. A UV-VIS detector was used to detect the separated components. Separation was performed in 2 types of capillaries, i.e. fused-silica and ethylene vinyl acetate copolymer (EVA) plastic capillaries. The concentration limit of detection (CLOD), mass limit of detection (MLOD), and relative standard deviations (RSD) of migration time and area were investigated. LOD was comparable to that of epi-fluorescence detection. LOD was lower when separation was performed in EVA capillary compared to fused-silica capillary. RSD of migration times of the porphyrins when separated in fused-silica capillary ranges between 0.5 to 1.6% and in EVA capillary ranges between 0.3 to 1.2%. Area RSD in fused-silica capillary ranges between 7 to 20% and in EVA capillary ranges between 4.7 to 12.3%. Urine spiked with porphyrins was also analysed by CE using fused-silica and EVA capillaries. Analysis of urine sample spiked with porphyrins showed that stacking effect of porphyrins was observable only in fused-silica and not in EVA capillary. This led to similar LOD in fused-silica capillary to those for EVA capillary. PMID- 9413618 TI - Testing human blood for cannabis by GC-MS. AB - A method for cannabis testing in human whole blood is presented. The procedure involves extraction of a 2 mL specimen acidified with 0.2 mL of 10% acetic acid in a silanized vial in presence of deuterated internal standards into hexane:ethyl acetate. After evaporation to dryness the drug and its metabolite were derivatized by methylation with iodomethane in tetrabutylammonium hydroxide dimethyl sulphoxide (DMSO). The derivatives were extracted into isoocatane for analysis by gas chromatography-mass spectrometry (GC-MS). The limit of quantification was 1.0 and 0.5 ng/mL for delta 9-tetrahydrocannabinol (THC) and 11-nor-9-carboxy-delta 9-tetrahydrocannabinol (THC-COOH), respectively. PMID- 9413619 TI - Impact of cognitive rehabilitation therapy on neuropsychological impairments as measured by brain perfusion SPECT: a longitudinal study. AB - Three patients, with known brain injury and neuropsychological impairments, are followed through an individualized cognitive rehabilitation programme and post discharge from the treatment programme. Single Photon Emission Computed Tomography (SPECT) of the brain was employed to evaluate resting relative cerebral blood flow (rCBF) during the process of recovery from brain injury. All patients experienced significant improvements on measures of neuropsychological functioning and improvements in rCBF during this longitudinal study. The specific changes in rCBF appear to be related to the location of the patient's brain injury and strategies particular to cognitive rehabilitation therapy. Continued improvements in rCBF, functional abilities, and cognitive skills were documented in these three cases up to 45 months post brain injury. PMID- 9413620 TI - CT findings in persistent vegetative state following blunt traumatic brain injury. AB - The use of linear measurements in the analysis of CT scans of TBI patients was found to contribute to the understanding of brain damage and were correlated with outcome in severe traumatic close brain injured patients. The purpose of the present study was to analyse the data obtained by the linear measurements on CT studies of TBI patients who remained in persistent vegetative state following blunt head trauma. All 27 patients included in the study were reported to be neurologically normal prior to injury. Thirteen patients, 11 remaining in persistent vegetative state (responsive but unaware) and two who died, constituted the worst outcome group. Fourteen patients who regained consciousness, underwent multidisciplinary evaluation when their recovery reached a plateau and were ranked according to severity of residual symptoms and outcome. The degree of correlation with the overall vocational outcome parameter with the various radiological indices was calculated as the Spearman rank correlation coefficient, with correction for tied scores. Fisher's z transformation was used to combine results with those of our previous analysis. Three radiological parameters showed a statistically significant correlation with clinical outcome. These were the right and left septum-caudate distance and the cerebroventricular index 2; these showed Spearman rank coefficients of 0.52, 0.45 and 0.48; with two tailed p-values under 0.01, 0.02 and 0.01 respectively. The width of the third ventricle suggested correlation with the clinical scoring. The findings of the present study point to the importance of loss of deep gray matter of the caudate nuclei and widening of the adjacent part of the lateral ventricles in catastrophic brain injury. This finding may highlight the role of localized ischemic changes, in addition to diffuse axonal injury. Values of over 8 mm for the width of the third ventricle and over 11 mm for septum caudate distance are suggestive of catastrophic and poor prognosis for recovery. PMID- 9413621 TI - Neurobehavioral consequences of closed head injury in the elderly. AB - The assessment of neurobehavioural outcome after head injury in older patients (> 60 year old) has met with difficultives, due to the obstacles in finding subjects who would constitute an appropriate control group. In the present study, survivors of closed head injury (CHI) of this age group were compared to two control groups: (1) orthopaedic patients (OP) who were injured in similar circumstances but did not sustain head injury and (2) healthy, age-matched volunteers (HC). Compared with HC, CHI and OP were impaired on word fluency, memory and reasoning. No differences were found between CHI and the OP. These results may indicate that, rather than resulting only from the head injury brought about by falling, the cognitive decline may predate the injury and increase the risk of accidents in old age. PMID- 9413622 TI - Antecedent management and compliance training improve adolescents' participation in early brain injury rehabilitation. AB - Children and adolescents with serious brain injuries are surviving in greater numbers, often entering rehabilitation settings while still emerging from coma. As the child's or adolescent's level of consciousness improves, increased demands to begin participating in self-care and therapeutic activities are presented. When the patient is unable to participate because of disorientation and agitation, the benefits of the rehabilitation admission may be jeopardized, limited rehabilitation resources may be prematurely exhausted, and behavioural sequelae often escalate. A non-concurrent multiple baseline across subjects experimental design was used to examine the effects of a behaviour management approach employing both antecedent environmental manipulations and operant conditioning-based compliance training. Data are presented on therapy attendance, disruptive behaviour, and agitation ratings across three recently brain-injured adolescents as therapeutic demands were gradually introduced. In all three cases, therapy attendance stabilized at a high level, disruptive behaviour decreased, and agitation was maintained at a moderate to low level. The results demonstrate the importance of carefully controlling the environment and coordinating the introduction of both therapeutic demands and positive reinforcement contingencies during early recovery from brain injury. PMID- 9413623 TI - The long term effects of traumatic brain injury on the roles of caregivers. AB - This study was conducted to identify the long term effects of traumatic brain injury (TBI) on the roles of caregivers. The subjects consisted of 155 caregivers of survivors with TBI who were randomly selected from 15 midwestern state brain injury association databases. A questionnaire was developed by the researchers to determine factors affecting role changes of caregivers. The Role Checklist, by Barris, Oakley and Kielhofner, was also included with the questionnaire. Both were mailed to each selected caregiver and used for data gathering. The data obtained were analysed to determine existing trends in the data. Graphs were utilized to depict the trends that was identified. The following trends and conclusions established by this research include: (a) behavioural effects of the survivor with a TBI are associated with the number of role changes experienced by caregivers; (b) participation in support systems is associated with the number of role changes experienced by caregivers; and (c) caregivers who care for a person with a TBI in the home will show a larger number of role changes than those who do not provide direct care for a person with a TBI. PMID- 9413624 TI - Beyond one-bun, two-shoe: recent advances in the psychological rehabilitation of memory disorders after acquired brain injury. AB - Memory disorders are one of the most frequent types of cognitive impairment encountered in neurological populations. The more severe degrees of such impairment case major disability and handicap, and have a profound impact on a person's capacity to engage in independent living. To date, commonly used remediation strategies range from drills and practice, including computer-based tasks, to mnemonic techniques and memory notebooks. In general, these therapies have met with varying degrees of success. The last decade has seen exciting developments in remediation techniques for memory disorders, a number of which are based on implicit learning skills, as well as programmes tailored to an individual's unique pattern of deficits. The present paper provides an overview of this literature and discusses issues relating to their application in rehabilitation programmes. PMID- 9413625 TI - G-proteins in allergy. PMID- 9413626 TI - Modulating effects of cetirizine on PLA2 in vitro. PMID- 9413627 TI - The molecular behaviour of cetirizine. PMID- 9413628 TI - Adhesion molecules and H1-antihistamines. PMID- 9413629 TI - Investigating bradykinin-induced reactions in the skin through microdialysis. PMID- 9413630 TI - The impact of cetirizine on microbial and viral infections. PMID- 9413631 TI - The clinical efficacy of antihistamines in the upper and lower airway. PMID- 9413632 TI - The effects of cetirizine on the function of inflammatory cells involved in the allergic response. PMID- 9413633 TI - Prognostic significance of t(14;18) and bcl-2 gene expression in follicular small cleaved cell lymphoma and diffuse large cell lymphoma. AB - OBJECTIVE: To evaluate whether there is an association between the presence of t(14;18) and bcl-2 gene expression, on one hand, and clinical presentation and outcome, on the other hand, in patients with follicular small cleaved cell lymphoma (FSCCL) and diffuse large cell lymphoma (DLCL), in light of conflicting reports concerning the prognostic significance of these parameters. DESIGN: Retrospective cohort study and molecular analysis of archived tissue. SETTING: Tertiary care hospital. PATIENTS: Sixty-two patients with non-Hodgkin's lymphoma, of whom 33 (13 women and 20 men) had FSCCL and 29 (9 women and 20 men) had DLCL. Their ages ranged from 27 to 88 years. INTERVENTIONS: Presence of t(14;18) was determined using Southern blot analysis or polymerase chain reaction or both. The level of bcl-2 gene expression was determined by immunohistological analysis using a monoclonal mouse anti-human antibody (DAKO-Bcl-2, 124). OUTCOME MEASURES: Clinical stage of lymphoma at diagnosis and responsiveness to treatment, and the correlation between these clinical parameters and t(14;18) status and bcl-2 gene expression. RESULTS: There was no clear association between t(14;18) status and prognosis for either FSCCL or DLCL. In contrast, high bcl-2 expression clearly predicted a generally poor prognosis in patients with FSCLL (p = 0.0146) and indicated resistance to treatment in those with DLCL (p = 0.0853). CONCLUSION: In non-Hodgkin's lymphoma, level of bcl-2 gene expression may represent a useful, independent prognostic indicator to identify high-risk patients and choose specific therapeutic approaches. PMID- 9413634 TI - Metallothionein expression in prostatic carcinoma: correlation with Gleason grade, pathologic stage, DNA content and serum level of prostate-specific antigen. AB - OBJECTIVE: To determine the expression of metallothionein (MT) in prostatic carcinoma by immunohistochemical staining. Several lines of evidence have indicated that MT may play a role in carcinogenesis and in drug resistance of tumours. DESIGN: Retrospective pathologic study. INTERVENTIONS: Formalin-fixed, paraffin-embedded archival tissues from 39 radical prostatectomies were analysed. All tumour foci were stained by ABC technique using a primary polyclonal rabbit antibody against rat liver MT. The staining intensity for MT was graded on a scale of 0 to 2+, and the histologic grading was done by the scheme of Gleason. OUTCOME MEASURES: Correlation of MT expression with Gleason grade, preoperative serum prostate-specific antigen (PSA) levels, pathologic stage and DNA content, including S-phase fraction (SPF) and proliferative index (PI). RESULTS: Most of the epithelium of normal prostate tissue had patchy, intense MT staining. All the grade II tumours foci showed intense (2+) staining for MT, while all grade IV and V foci were persistently negative. The grade III tumours foci were heterogeneous. The MT-positive foci showed both nuclear and cytoplasmic staining of variable extent. There were 9, 15, 13 and 2 tumours with pathologic stage B, C1, C2 and D1, respectively. The serum PSA levels ranged from 1 to 16 ng/mL. No apparent correlation existed between the MT staining pattern and the pathologic stage or preoperative PSA level. Thirty-four of the tumours were diploid and 5 were tetraploid. There were significantly higher SPF and PI mean values in the MT stained tumour cells (p < 0.05), suggesting that MT preferentially stains an epithelial subpopulation, possibly the proliferating cell compartment. CONCLUSION: The positive correlation of MT expression with Gleason grade in prostatic adenocarcinoma suggests a possible role for MT in oncogenesis in prostate cancer. PMID- 9413635 TI - Adherence to clinical guidelines for the therapeutic management of HIV disease. AB - OBJECTIVE: To determine the knowledge of HIV-disease management and the adherence to contemporary guidelines among British Columbia physicians whose practices focused on HIV/AIDS. DESIGN: Self-administered mail survey. PARTICIPANTS: All 659 physicians registered in a province-wide HIV/AIDS drug treatment program. OUTCOME MEASURES: Data on demographic and personal characteristics of respondents, level of HIV-related experience, use of preventive vaccinations and tests, and preferred approaches to the prophylaxis and treatment of common opportunistic infections. Knowledge scores in 4 areas of patient care, as well as an overall score, were computed by comparing respondents' answers with the therapeutic strategies recommended at the time of the survey. Associations between physician characteristics and knowledge scores were identified by linear regression analysis. RESULTS: Of the 659 physicians surveyed, 65% returned responses: only 38% returned completed surveys while a further 27% returned a follow-up survey that asked nonrespondents about their demographic characteristics and HIV-related experience. Scores for specific areas of patient management ranged from 29% for the treatment of opportunistic infections to 62% for preventive measures, with a mean overall score of 47%. Physician knowledge in all areas of patient care was associated with the number of HIV-positive patients in the practice (p = 0.003 to p < 0.001). Physicians who were younger were more knowledgeable regarding preventive measures (p = 0.001); those whose practice location was in Vancouver had a greater knowledge of prophylaxis (p = 0.047); and those who had medical specialty training were more knowledgeable about the treatment of opportunistic infections (p = 0.009). CONCLUSIONS: There is substantial disparity in how physicians approach the management of HIV and related conditions. Deviations from therapeutic guidelines are common and may be associated with physician characteristics, particularly lack of experience in managing HIV. PMID- 9413636 TI - Psychiatric evaluation and outcomes in candidates for heart transplantation. AB - OBJECTIVE: To describe the results of psychiatric evaluations and to describe psychiatric outcomes in patients in a heart transplantation program. DESIGN: Descriptive study. SETTING: Heart transplant unit at the Royal Victoria Hospital in Montreal from September 1984 to December 1995. PATIENTS: A total of 706 candidates for heart transplantation, of whom 226 received a graft. INTERVENTIONS: All candidates underwent a psychiatric evaluation consisting of a semi-structured interview with mental status examination and diagnosis of mental disorders. OUTCOME MEASURES: Results of psychiatric evaluation and postoperative recovery and complications. RESULTS: Twenty-eight candidates were found to be unsuitable for surgery because of psychiatric illness. The heart transplant recipients who had previously suffered from a psychiatric disorder fared worse than those who had not. The psychiatrist's recommendations for or against surgery depended on the patient's ability to cope with a number of stressors, including compliance with the medical regimen, the wait for a donor, the surgical procedure itself, adaptation to life with a new organ and the resolution of a distressing emotional state. Postoperative psychiatric complications ranged from organic mental syndromes to depression. CONCLUSIONS: With the increase in the number of heart transplantations, the competition for organ donors is intensifying, and patient selection requires greater involvement of psychiatrists. PMID- 9413637 TI - Change and the academic health science centre: a 1997 perspective. PMID- 9413638 TI - Investigator-initiated research: a necessity or a luxury? PMID- 9413639 TI - Professionalism, laws and kings. PMID- 9413640 TI - As the world turns: the consequences for health. PMID- 9413641 TI - Information technology and the future of medical education. PMID- 9413643 TI - A methodological study of arterial wall function using ultrasound technique. AB - We aimed to establish reference values for three important properties of the arterial wall using a conventional ultrasound scanner. We measured: (1) intima media thickness (IMT) of the carotid arteries with the internal trace function of the ultrasound system; (2) wall stiffness by pulsatile diameter changes in the right common carotid artery assessed by M-mode; and (3) endothelial function expressed as flow-mediated dilatation (FMD) of the brachial artery. IMT and wall stiffness measurements and reproducibility were compared with those obtained by external analysing systems. All variables were obtained in healthy subjects (n = 20), 29-53 years old. IMT increased with age (P < 0.01). There was no difference in IMT between men and women. The inter-operator variability for measuring IMT was 6-9%. The same order of reproducibility was obtained with an external PC based analysing system. Regarding wall stiffness, no correlation was found with age, nor any difference between men and women. A low intra-operator variability (CV < 10%) was found for measurements of wall stiffness with both M-mode and an external wall tracking system. FMD of the brachial artery diminished with age (P < 0.01). There was a relation between FMD and brachial artery size (P < 0.01) and, therefore, as men have larger arterial diameters (P < 0.01), smaller FMD in men. We conclude that it is possible to characterize arterial wall function non invasively in an adequately reproducible manner using a conventional ultrasound system in healthy middle-aged men and women. PMID- 9413642 TI - Scintigraphic assessment of focal platelet accumulations following infrainguinal bypass surgery in humans. AB - Enothelial injury is assumed to be of pathogenetic significance in the development of graft stenoses, which remain a major cause of failure of peripheral bypasses. The aim of this study was to assess endothelial injury related to infrainguinal bypass surgery by indium-111 platelet scintigraphy. In 28 patients undergoing in situ vein (n = 24), composite vein polytetrafluoroethylene (PTFE) (n = 1) or PTFE (n = 3) bypass surgery, assumed vascular injuries were recorded intraoperatively. Autologous indium-111-labelled platelets were injected into the inflow artery immediately after restoration of flow in the graft. Platelet deposition was assessed by gamma-camera images of thigh and crus obtained 4 and/or 24 h after surgery. Areas of focally increased activity were recorded and graded as moderate or intense. In the 24 vein bypasses, a median of two (range 0-5) areas of focally increased radioactivity were seen at the proximal anastomosis (n = 21), in the body of the graft (n = 20) or at the distal anastomosis (n = 9). The activity was moderate in 27 cases and intense in 23 cases. Scintigraphic evidence of focal platelet aggregation in vein grafts was not correlated with preoperative antiplatelet therapy or vein graft diameter. Only 2 of the 20 intragraft platelet depositions occurred in areas where intra-operative vascular injury was suspected. In the composite graft and the PTFE grafts, diffuse activity was observed throughout the entire bypass. In conclusion, focal activity accumulations, suggesting localized endothelial injury, were observed in the majority of in situ vein bypasses, in particular at the sites of the anastomoses. Prosthetic bypasses were characterized by diffuse platelet aggregation. PMID- 9413644 TI - Stiffness of the common carotid artery in healthy 50-year-old subjects. AB - We examined whether, in a group of healthy 50 year-old subjects, there was a relation between the stiffness of the common carotid artery and risk factors for atherosclerosis and, further, if there was a sex difference in these healthy individuals. A group of 248 healthy subjects (123 men and 125 women), all 50 years of age, were examined. The elastic properties of the common carotid artery, measured in terms of stiffness index, were investigated with an ultrasound technique. Men had significantly higher stiffness index than women (5.46 +/- 1.32 vs. 5.04 +/- 1.05, P < 0.01). The arterial diameter was significantly larger in men. When comparing stiffness index in a subgroup of 38 men and women with similar arterial diameters, no significant difference in stiffness index was found. Multivariate analysis showed that smoking habits, measured as pack-years, insulin levels and the low-density lipoprotein (LDL)-/high-density lipoprotein (HDL)-cholesterol ratio was independently significantly associated with the stiffness index. Sex did not emerge as a significant variable. In conclusion, there is a relation between the stiffness of the carotid arteries and risk factors for atherosclerosis such as smoking, insulin levels and LDL-/HDL cholesterol ratio in healthy 50-year-old subjects. The stiffness is higher in men than in women, probably dependent on differences in arterial size and possibly also on risk factors. PMID- 9413645 TI - Association of carotid arterial distensibility with Doppler indices of left ventricular filling in 50-year-old subjects. AB - Doppler indices of left ventricular diastolic filling are associated with various cardiac and extracardiac factors. Afterload is one of the extracardiac factors influencing left ventricular diastolic filling. The distensibility of the great arteries is one of the components of afterload. In this study, the relation between Doppler indices of left ventricular filling and the distensibility of the common carotid arteries was investigated. We studied 237 subjects at 50 years of age with Doppler echocardiography and ultrasound examination of the common carotid arteries. The following Doppler indices of left ventricular filling were studied: peak early diastolic velocity E-wave, peak atrial diastolic velocity A wave and early to atrial peak velocity ratio, E/A. Carotid arterial characteristics were: distensibility coefficient, carotid arterial diameter change in systole and fractional change in the carotid arterial diameter. The relation between Doppler indices of left ventricular filling and carotid arterial characteristics was assessed by univariate and multivariate regression analysis. There was a significant univariate, positive association between E/A ratio and carotid arterial distensibility (r = 0.27, P < 0.001), carotid arterial systolic diameter change (r = 0.19, P < 0.005) and fractional change of the carotid arterial diameter (r = 0.20, P < 0.005). In multivariate analysis, E/A ratio was independently associated with carotid arterial distensibility (P < 0.005), after adjusting for heart rate, body mass index and gender. Decreased carotid arterial distensibility was associated with a reduction in E/A ratio, suggesting that arterial distensibility may have an effect on left ventricular diastolic filling or that changes in the arterial elastic properties are associated with corresponding structural changes in the left ventricle. PMID- 9413646 TI - Kidney size estimation in piglets using dimercaptosuccinic acid (DMSA) scintigraphy. AB - The objective of this study was to validate kidney size measurements obtained by DMSA scintigraphy in kidneys of sizes relevant in paediatric practice. Kidney length, width and volume were measured by DMSA scintigraphy in 10 piglets in vivo and compared with measurements on excised kidneys using callipers, planar scintigraphy and submersion in water. In addition, a kidney phantom was constructed and placed in a torso phantom. The length, width and volume of the kidney phantom were compared with planar and tomographic scintigraphic measurements. The mean overestimation of kidney length by scintigraphy was 1.7% (+/- 1.2%) on excised kidneys and 3.9% (+/- 2.4%) in vivo. The corresponding figures for width measurements were 7.0% (+/- 3.9%) and 7.8% (+/- 4.4%). Tomography volume estimation overestimated the submersion volume by 23% (+/- 12%). Scintigraphic measurements on the phantom over-estimated length by 5%, width by 8% and volume by 11%. Scintigraphic renal length measurements are reliable and differ from the anatomical length by < 4% compared with excised kidneys and < 8% in vivo. Scintigraphic width measurements and volume estimations, especially on small kidneys, are less reliable. PMID- 9413648 TI - Influences of mandatory breathing on rhythmical components of electrodermal activity. AB - We investigated whether rhythmical components of electrodermal activity (EDA) can be influenced by voluntary modification of the respiratory frequency. Fifty-five volunteers participated in an experiment with nine mandatory sections of cycle times between 3.1 s and 12.2 s. The cycle time of the rhythmical EDA components often differed considerably from respiratory cycle time. ANOVA yielded that both cycle time and position within the section had an effect on the EDA power of the mandatory cycles (P < 0.05). No significant interaction between effects of section and of position was discovered (P > 0.1). A separate analysis of the cycle times of the rhythmical EDA components revealed integer relationship with the respiratory cycle times. Thus, voluntary changes in the respiratory frequency can influence occurrence and cycle time of rhythmical EDA components in a way similar to sliding coordination in the sense of E. von Holst. PMID- 9413647 TI - Reproducibility of a semiautomated acetylene rebreathing technique for measuring cardiac output in humans at rest. AB - The specific aims of the present study were to determine: (1) the day-to-day reproducibility of a semiautomated acetylene rebreathing technique for measuring cardiac output under resting conditions; (2) the reproducibility of this technique among subjects differing in gender and age; and (3) the number of trials within a session necessary to maximize the day-to-day reproducibility of the technique. To address these aims, cardiac output was measured in 21 healthy men (n = 8) and women (n = 13) between the ages of 25 and 71 years in the supine posture on two separate days. Mean levels of cardiac output at rest were similar on day 1 vs. day 2 in the overall group. Cardiac output measured on day 1 was highly correlated (r = 0.98, P < 0.001) with cardiac output measured on day 2. The day 1 to day 2 mean difference in cardiac output for the individual subjects was < 4%. The mean levels of heart rate and stroke volume also were similar between day 1 and day 2. The relation between cardiac output measured on day 1 vs. day 2 in the gender and age subgroups was similar to that observed in the overall group. The mean absolute difference among the three rebreathing trials within a day was 360 ml min-1 in the overall group, with a coefficient of variation of 7%. The variability between rebreathing trials measured on day 1 vs. day 2 in the gender and age subgroups was similar to that observed in the overall group. The reliability of cardiac output measured on different days was excellent with a single rebreathing trial (r = 0.93) and improved significantly up to three trials (r = 0.98). In conclusion, the findings of the present study indicate that the acetylene rebreathing technique can be a highly reproducible method for measuring cardiac output under resting conditions. The reproducibility is consistently strong in healthy humans of varying age and in both genders, and is enhanced by the use of multiple trials. PMID- 9413649 TI - Video fluorescein imaging of the skin: description of an overviewing technique for functional evaluation of regional cutaneous blood perfusion in occlusive arterial disease of the limbs. AB - In peripheral arterial occlusive disease (PAOD), and particularly in critical limb ischaemia, fluorescence recording of the skin after i.v. injection of sodium fluorescein provides an overviewing functional imaging of regional blood perfusion as revealed by a continuous, almost geographical, mapping of changes in the dynamic fluorescence pattern. Based on the previously used technique of rapid sequence flash photography introduced by Lund and Lund in 1973, a new video fluorescence imaging (VFI) technique has been developed and used in clinical routine. VFI has a broad versatility with non-touch access to areas of different size, such as the feet, hands or larger limb areas. Examples of application are the diagnosis of small artery disease, predictive prognosis of critical limb ischaemia, evaluation of distal outflow in connection with arterial reconstruction or pharmacotherapy of non-reconstructable critical ischaemia, as well as in decisions on amputation and assessment of amputation level. Thanks to its overviewing qualities with reproducible mapping of blood perfusion in standardized examination conditions, VFI would seem to have a well-justified role in angiology and above all in PAOD in both research and clinical routine work. PMID- 9413650 TI - Influence of body size and age on mitral ring motion. AB - Left ventricular systolic and diastolic function can be assessed by studying mitral ring motion. Reference values for a wide range of age and body size are lacking however. The motion of the mitral ring was studied with M-mode echocardiography using the apical four- and two-chamber views. Data are reported as the average of measurements of four points on the mitral ring. Data were analysed using the stepwise multiple regression technique, with age, gender, height, weight, body surface area and heart rate as independent variables. A total of 70 healthy subjects were studied. In children and adolescents under age 18, there was strong correlation between mitral ring motion and body size, age and heart rate. The ring motion was best described as mitral ring motion (mm) = 2.2 + 0.078 x height (cm) (SEE = 1.0 mm). In adults, mitral ring motion was correlated with age, height and heart rate but not with weight or body surface area. Ring motion could be described from the following equations: mitral ring motion (mm) = 12.7 - 0.060 x age (years) + 0.031 x height (cm) (SEE = 1.2) or, if only age is taken into account, mitral ring motion = 18.4 - 0.065 x age (SEE = 1.2). In both children and adolescents up to age 18 and in adults, atrial contribution correlated significantly and positively to age and inversely to heart rate but not to height, weight, body surface area or gender. Atrial contribution was best described by the equation: atrial contribution = 0.15 + 0.0039 x age (SEE = 0.027). Thus, age and body size influences mitral ring motion and should be taken into account when interpreting patient data. PMID- 9413651 TI - Descriptive definition and historic aspects of sarcoidosis. AB - Sarcoidosis is set within the framework of a large family of granulomatous disorders, so it has many mimics. Granuloma formation is defined with its immunologic basis. The criteria of activity of sarcoidosis are included in the broadly based definition. Future knowledge will derive from basic immunology and molecular biology. Historic aspects are based on pioneers, who have died, and their seminal contributions. Evolution of knowledge derives from information gained at international conferences and from our biennial journal, "Sarcoidosis, Vasculitis and Diffuse Lung Diseases." PMID- 9413652 TI - Global epidemiology of sarcoidosis. What story do prevalence and incidence tell us? AB - Textbooks of clinical medicine often begin with "epidemiology" of the disease by describing the distribution of patients' characteristics in terms of age, gender, race, and so on. As a result, many clinicians erroneously think the description of such distribution is a role only for epidemiology. The real role of epidemiology, however, is to search for the determinants of and ways to prevent the disease. In this article, the recent informative papers on the epidemiology of sarcoidosis are reviewed in the light of modern sarcoidology. PMID- 9413653 TI - Etiology of sarcoidosis. AB - Since sarcoidosis was first recognized as a distinct clinical entity, investigators have speculated that a transmissible agent may cause sarcoidosis. Recent attempts at directly isolating infectious organisms or indirectly detecting microbial DNA or RNA from sarcoid tissue have led to inconclusive results. Studies on the immunopathogenic origins of sarcoidosis have provided evidence of persistent antigenic stimulation at sites of inflammation that are associated with dysregulated cytokine production. To date, however, the challenge of defining the cause of sarcoidosis remains unmet. PMID- 9413654 TI - Genetics of sarcoidosis. AB - Hereditary susceptibility to sarcoidosis is suggested by ethnic preponderance, familial clustering, and multigenerational involvement. The genetics of sarcoidosis cannot be adequately addressed in small samples of patients; a large scale study with stratification for patient phenotypic differences is necessary. A study that uses both genetic marker and environmental data would be able to control for and examine different causative mechanisms. Until such a well designed, comprehensive study is carried out, we are left with interesting patterns of disease in families and uncertain allelic associations. PMID- 9413655 TI - Immunology of sarcoidosis. AB - Because of its association with cutaneous anergy, sarcoidosis was originally viewed as a defect of cellular immunity. Supporting that misperception were early studies of peripheral blood lymphocytes that found lymphopenia and impaired lymphocyte responses to mitogens and recall antigens. The clue to a vast underlying network of complex hyperactive cellular immune functions was discovered in the paradoxical finding of in vitro spontaneous lymphoblastic transformation and lymphokine production. Subsequently, investigative focus shifted to the activated, proliferating T-helper lymphocytes, the lymphokines of which were found to function in the recruitment and retention of monocytes for granuloma development. T-helper lymphocytes also contributed to the mechanism of hypergammaglobulinemia through their influence on B cells. The most intriguing question about sarcoid immunology is the initiating factor that triggers the T lymphocyte activation and proliferation in the first place. There is much to suggest that antigen processing and presentation launches the process. Because lymphocyte activation and proliferation antedate granuloma formation at K-S skin test sites and in the lung, we combined the harvesting technique of BAL with the K-S bioassay to show that granulomagenic antigen is being processed by monocyte macrophages. The finding of autologous monocyte-macrophage granulomagenicity raises the distinct possibility that sarcoidosis is a unique cell-mediated type of autoimmune process. The isolation and identification of the granulomagenic factor is the exciting research frontier ahead. PMID- 9413656 TI - Pathology of sarcoidosis. AB - This article reviews the pathology of sarcoidosis that covers the general and systemic aspects of the disease. Macroscopic and microscopic descriptions of the disease process are given for selected organs. PMID- 9413657 TI - Pulmonary sarcoidosis. AB - Sarcoidosis involves the bronchi or lung in more than 90 percent of patients. Intrathoracic manifestations are protean, ranging from asymptomatic bilateral hilar lymphadenopathy to chronic, progressive, (ultimately fatal), respiratory insufficiency. The clinical course is highly variable, and optimal management and treatment are controversial. We review the salient radiographic, physiologic, and histopathologic features of pulmonary sarcoidosis and discuss rare intrathoracic complications (e.g., bronchostenosis, mycetomas, nodular sarcoidosis, necrotizing sarcoid angiitis and granulomatosis, pulmonary vascular and pleural involvement). We discuss the chest radiographic staging system and the role of ancillary diagnostic modalities including high resolution thin section computed tomographic scans (HRCT), bronchoalveolar lavage, radionuclide scan, and serum angiotensin enzyme converting enzyme. Indications for therapy and an overview of therapeutic options are outlined. PMID- 9413658 TI - Bronchoalveolar lavage. Still useful in diagnosing sarcoidosis? AB - BALF parameters, if evaluated as a diagnostic or prognostic tool, should be based on a full understanding of the clinical profiles and course of pulmonary sarcoidosis. The various markers that have been reported so far are unreliable in determining the prognosis, although BALF lymphocytes and CD4/CD8 ratios are still useful for diagnosing sarcoidosis. It is critical to find feasible markers that relate to a change in disease activity and prognosis, because markers of chronicity may be different from those of progressively worsening sarcoidosis. PMID- 9413659 TI - Nuclear imaging. 67Gallium, 201thallium, 18F-labeled fluoro-2-deoxy-D-glucose positron emission tomography. AB - 67Gallium scan has been used for years in sarcoidosis as a marker of activity, a determiner of the extent and distribution of the disease, a diagnostic support, and an aid in therapeutic management. Because of its limited sensitivity and specificity for sarcoidosis, however, it is currently used mainly to assist in diagnosis in difficult cases, particularly in those with isolated extrathoracic sarcoidosis. The finding of the typical lambda or panda patterns supports the diagnosis and reinforces the indication to perform an appropriate biopsy or Kveim Siltzbach test. In addition, the detection of clinically silent extrathoracic uptake may provide sites for biopsy. 67Gallium scans' routine use in the follow up of pulmonary sarcoidosis under treatment has decreased because that is best accomplished by means of serial chest radiographs and PFT. 201Thallium scintigraphy studies the myocardial perfusion and is complementary to echocardiography and 24-hour electrocardiographic monitoring in the assessment of sarcoid cardiac involvement. It typically shows segmental areas of decreased uptake in the ventricular myocardium that disappear or decrease in size during stress or after intravenous administration of dipyridamole. That reverse distribution is not specific for cardiac sarcoidosis, however, because it may also occur in other cardiomyopathies. PET is based on the increase of glucose metabolism in inflamed tissues. It may have great potential to assess sarcoidosis activity, but it is still largely experimental and is not routinely employed. PMID- 9413660 TI - Cardiac and neurologic dysfunction in sarcoidosis. AB - Clinically apparent involvement of the heart and nervous system occurs in a relatively small number of patients with sarcoidosis. The diagnosis of myocardial and neurological sarcoidosis is difficult because anatomic presence of granulomas without clinical dysfunction is an important feature of sarcoidosis. The chest radiography is abnormal in 8 of every 10 patients with myocardial or neurosarcoidosis. Serum angiotensin-converting enzyme and gallium uptake studies may provide some indication of the extent and severity of the granulomatous process. Corticosteriods are the mainstay of therapy but chloroquine or hydroxychloriquine, methotrexate, and azathioprine are also effective. Prognosis of myocardial and neurological sarcoidosis is poor. PMID- 9413661 TI - Biochemical changes in sarcoidosis. AB - Biochemical markers in sarcoidosis are closely related to the immunological events and the activity of inflammatory effector cells at sites of granuloma formation. The markers can be measured in serum, then reflecting whole body concentration, or in BAL fluid, then indicating activity in the lung. Only calcium and ACE serum levels have gained a proven value in the clinical field. PMID- 9413662 TI - Treatment with corticosteroids. AB - Oral corticosteroids remain the cornerstone therapy for sarcoidosis. Critical clinical decisions include selecting the patient who should be treated, dose and duration of therapy, and accurate analysis of the anticipated benefits and potential side effects for the individual patient. The treatment of pulmonary and cardiac sarcoidosis is emphasized and the role of inhaled corticosteroids in the treatment of pulmonary sarcoidosis is reviewed. PMID- 9413663 TI - Steroid-sparing alternative treatments for sarcoidosis. AB - Alternatives to corticosteroids for the treatment of sarcoidosis are reviewed. These include cytotoxic agents such as methotrexate, azathioprine, and cyclophosphamide. In addition, agents such as hydroxychloroquine and cyclosporine are reviewed. The efficacy, toxicity, and timing of these drugs in the management of sarcoidosis is discussed. PMID- 9413664 TI - Role of transplantation (lung, liver, and heart) in sarcoidosis. AB - Organ transplantation is an option for sarcoidosis patients with end-stage lung, liver or heart disease. Survival statistics vary for the organ transplanted but are not too different from survival rates for other systemic disorders. Although infection and rejection are troublesome for all organ recipients including those with sarcoidosis, there is the added problem of recurrence of sarcoidosis in the allograft. Sarcoidosis is not an absolute contraindication for organ transplantation for the majority of transplantation centers. PMID- 9413665 TI - The impact of the Massachusetts Managed Mental Health/Substance Abuse Program on outpatient mental health clinics. AB - Medicaid managed care initiatives pose special challenges to outpatient providers. During the first two full years of the Massachusetts Mental Health/Substance Abuse initiative, an analysis of cost and utilization data showed that outpatient mental health utilization and expenditures dropped slightly, although far less than did expenditures and utilization for inpatient facilities. In a telephone survey of a stratified random sample of outpatient providers, they reported that access, appropriate utilization, quality of care, the severity of their clients and aftercare coordination increased, while length of stay for these clients decreased. In their clinical practices, agencies shifted toward more emphasis on group and family care and brief therapies. As organizations, they made substantial operational changes. As a result, some agencies did better, while others did worse, under this new system. PMID- 9413666 TI - Depression in the caregiving mothers of adult schizophrenics: a test of the resource deterioration model. AB - The predictive validity of the resource deterioration model was tested with a sample of 100 Black, elderly, low-income, unmarried, caregiving mothers of adult schizophrenic sons and/or daughters. Stressors consisted of three social variables (burden of care, economic strain, undesirable life events) and one physical variable (poor physical health). Stress mediators consisted of coping and social support resources; the outcome variable was defined as depression. The results indicated support for the resource deterioration model with regard to a physical stressor and coping resources, but not for social stressors and social support resources. PMID- 9413667 TI - Fathers of persons with mental illness: a preliminary study of coping capacity and service needs. AB - Issues faced by fathers coping with the mental illness of an adult child represent an unexplored dimension of service needs. A preliminary exploratory study found that a group of 25 such fathers manifested important indicators of emotional stress that were largely unrecognized and unacknowledged. They also demonstrated typical patterns of healing that were different from those experienced by their wives. The paper reports findings that suggest that fathers employ more isolating strategies, and suggests service approaches that might be more fruitful in responding to these serious needs. PMID- 9413668 TI - Styles of case management: the philosophy and practice of case managers. AB - A great deal of discussion and research has gone into defining and clarifying the role of "case manager" (CM) for persons with severe mental illness. This three state survey examines the philosophy and activities of practicing CMs in an attempt to identify current styles of case management. A cluster analysis based on CM rankings of five CM functions suggested four styles of case management: "supportive social workers", "individual therapist", "therapist broker", and "community advocate". Overall, CMs rated supportive interventions as most important and formal psychotherapy as relatively unimportant. CM style was related to CM activity (i.e., distribution of effort). Differences between states are noted and implications for future research are discussed. PMID- 9413669 TI - Substance use among the mentally ill: a comparison of Italian and American samples. AB - Thirty-six patients with long-standing psychotic illness randomly selected among attenders at two community centers of Bologna were interviewed as were their therapists, in order to determine prevalence and patterns of substance use, reasons for use and effects on illness. Protocols were the same as used in previous research carried out in Boulder, Colorado. For all substances, except stimulants and over-the-counter products, prevalence rates were lower in the Bologna sample than in Boulder. Overall lifetime rates of substance use were significantly lower in the Bologna sample. Nevertheless 47.2% of the Bologna sample had a lifetime moderate to severe drug use and nearly two-thirds of the sample had a current drug use of some severity. Reasons for use equally reflected attempts to alleviate unpleasant emotional states and recreational purposes. Patients with high degrees of psychopathology had significantly lower scores on severity of drug use. PMID- 9413670 TI - A state-university collaboration to serve persons with developmental disabilities and mental health needs. AB - Persons with developmental disabilities and mental health needs present a complex and potentially rewarding challenge to workers in both developmental disabilities (DD) and mental health (MH) fields. These individuals may be shuttled between (DD) and (MH) systems. Effective care requires the person obtain services from both systems. This paper describes one possible mechanism of service delivery for persons with developmental disabilities and mental health needs. PMID- 9413671 TI - Behavioral thermoregulation in chicks: the best nest. AB - The ability of prehomeothermic chicks to thermoregulate behaviorally was studied in chicks with continuous access to heated nests, running wheels, and separate sources of high and low protein. In Experiment 1, cold-reared groups with heated or unheated transparent nests ate the same amount and selected the same dietary fractions, but chicks with heated nests ran less and grew faster. Despite this, groups maintained normal body temperatures. In Experiment 2, chicks were cold- or warm-reared with heated or unheated painted nests, or no nests. Cold-reared chicks with heated nests spent most of their time in them. They selected diets containing a higher protein:carbohydrate ratio than cold-reared chicks with unheated nests but ate less, thereby consuming less absolute protein and growing more slowly. Despite differences in growth, intake, and dietary choice, all chicks maintained normal body temperatures. These data reveal that behavioral thermoregulation has a privileged status for chicks over the first 3 weeks of life. Prehomeothermic chicks exercise complex and effective solutions to energetic challenges when offered behavioral options that simulate those available in nature. PMID- 9413672 TI - Conditioned place preference in 12-day-old Japanese quail. AB - Four experiments assessed the ability of 12-day-old Japanese quail to learn a conditioned place preference (CPP). In Experiment 1, immature quail learned to prefer a place paired with normal food over a place paired with tainted food. Experiment 2 indicated that this kind of learning can be achieved with as few as 2 days of training. It was discovered in Experiment 3 that place preferences can be established with exposure to only one hedonic event. Quail learned to prefer a chamber paired with either normal food or tainted food over a chamber that did not contain a hedonic stimulus. Experiment 4 successfully replicated the 2-day normal-food-induced place preference in the previous experiment, while also showing that mere context exposure is not sufficient to produce CPP. PMID- 9413673 TI - Five generations of selective breeding for ultrasonic vocalization (USV) responses in N:NIH strain rats. AB - This article reports on early results from an ongoing selective breeding study in which rats were selected for different rates of ultrasonic vocalization (USV) in response to isolation. Using the N:NIH strain, all litters were screened at 10 (+/- 1) days of age in a 2-min isolation test, and those males and females with the highest (or lowest) rates in each litter were selected for later breeding with like breeders from unrelated litters. A Random line (unselected control) was also bred. In the first selected generation (S1), the Low line diverged from Random line controls, and has maintained significantly lower rates over all generations since. In the S3 generation, the High line diverged significantly from Random line controls, and has shown significantly higher USV rates in each succeeding generation. No line differences were found in other behaviors measured in isolation. Data from a small sample of S5 pups tested at postnatal Days 3, 10, 14, and 18 days showed that individual pups' rates of USV from Day 10 predicted those at Day 14, consistent with findings from an unselected generation. Ambient temperature, modulated by body weight, controlled USV at Day 3, whereas at Days 10 and 14 line accounted for most of the variance in USV. This is the first instance of laboratory selection occurring on the basis of an infantile trait. PMID- 9413674 TI - The development of eye movements in the zebrafish (Danio rerio). AB - We investigated the development of oculomotor activity in zebrafish embryos and larvae of ages 48-96 hrs postfertilization (hpf). The optokinetic response (OKR: smooth tracking movements evoked by a rotating striped drum) improved steadily after its onset at 73 hpf, and by 96 hpf had a achieved a gain (eye velocity/drum velocity) of 0.9, comparable to adult performance. Reset movements (the fast phase of optokinetic nystagmus) developed over 75-81 hpf. The vestibuloocular reflex (VOR: compensatory eye movements evoked by passive rotation of the head) developed over 74-81 hpf, and the associated reset movements, over 76-81 hpf. The VOR was qualitatively normal in dark-reared fish, which excludes an essential role for visual experience in its early development. Spontaneous saccadic movements (the fast shift of eye position) appeared between 81 and 96 hpf, and at 96 hpf had maximum velocities that were comparable to adults. These results are compared to, and found to be incompatible with, two earlier ideas of motor development: behavioral "differentiation" and "encephalization." PMID- 9413675 TI - Mother and infant smiling exchanges during face-to-face interaction in infants with and without Down syndrome. AB - We examined social smiling in infants with and without Down syndrome, aged from 3.2 to 13.6 months old. They were videotaped during an episode of spontaneous face-to-face interaction and a subsequent mother's still-face situation. Results indicated that infants smiled longer in the spontaneous face-to-face episode than in the still-face episode, even though this result was only significant in typically developing infants. Typically developing infants also smiled for a longer period than Down's syndrome infants during the spontaneous interaction episode. Moreover, infant's smile preceded the onset of the mother's smile, but in 6.2- to 13.6-month-old typically developing infants, the probability of mothers smiling before infants increased. These findings emphasize the possible existence of differences in the development of facial expression from signs to social symbols between infants with and without Down syndrome. PMID- 9413676 TI - Individual differences in object permanence performance at 8 months: locomotor experience and brain electrical activity. AB - This work was designed to investigate individual differences in hands-and-knees crawling and frontal brain electrical activity with respect to object permanence performance in 76 eight-month-old infants. Four groups of infants (one prelocomotor and 3 with varying lengths of hands-and-knees crawling experience) were tested on an object permanence scale in a research design similar to that used by Kermoian and Campos (1988). In addition, baseline EEG was recorded and used as an indicator of brain development, as in the Bell and Fox (1992) longitudinal study. Individual differences in frontal and occipital EEG power and in locomotor experience were associated with performance on the object permanence task. Infants successful at A-not-B exhibited greater frontal EEG power and greater occipital EEG power than unsuccessful infants. In contrast to Kermoian and Campos (1988), who noted that long-term crawling experience was associated with higher performance on an object permanence scale, infants in this study with any amount of hands and knees crawling experience performed at a higher level on the object permanence scale than prelocomotor infants. There was no interaction among brain electrical activity, locomotor experience, and object permanence performance. These data highlight the value of electrophysiological research and the need for a brain-behavior model of object permanence performance that incorporates both electrophysiological and behavioral factors. PMID- 9413677 TI - FDA perspective on specifications for biotechnology products--from IND to PLA. AB - Quality standards are obligatory throughout development, approval and post marketing phases of biotechnology-derived products, thus assuring product identity, purity, and potency/strength. The process of developing and setting specifications should be based on sound science and should represent a logical progression of actions based on the use of experiential data spanning manufacturing process validation, consistency in production, and characterization of relevant product properties/attributes, by multiple analytical means. This interactive process occurs in phases, varying in rigour. It is best described as encompassing a framework which starts with the implementation of realistic/practical operational quality limits, progressing to the establishment/adoption of more stringent specifications. The historical database is generated from preclinical, toxicology and early clinical lots. This supports the clinical development programme which, as it progresses, allows for further assay method validation/refinement, adoption/addition due to relevant or newly recognized product attributes or rejection due to irrelevance. In the next phase, (licensing/approval) specifications are set through extended experience and validation of both the preparative and analytical processes, to include availability of suitable reference standards and extensive product characterization throughout its proposed dating period. Subsequent to product approval, the incremental database of test results serves as a natural continuum for further evolving/refining specifications. While there is considerable latitude in the kinds of testing modalities finally adopted to establish product quality on a routine basis, for both drugs and drug products, it is important that the selection takes into consideration relevant (significant) product characteristics that appropriately reflect on identity, purity and potency. PMID- 9413678 TI - Global perspective on specifications for biotechnology products--perspective from Japan. PMID- 9413679 TI - Development of specifications for biotechnology products--perspective from Europe. PMID- 9413680 TI - Specifications from a biotechnology industry perspective. AB - The emergence of new analytical technology and the production of pharmaceuticals for a global market in a cost-effective manner necessitate the establishment of worldwide specifications that are appropriate for the product and the manufacturing process. This requires a thorough knowledge of the protein and control of the systems that produce it as well as an understanding of the accuracy and precision of the assays used for testing. Harmonization of specifications among the worldwide regulatory authorities is critical for the future development of new pharmaceuticals. A continuing dialogue between industry and regulators to achieve this goal needs to be encouraged and supported. PMID- 9413681 TI - Appropriate specifications at the IND stage. AB - Biopharmaceutical product specifications are to be "scientifically sound and appropriate". However, how does a biopharmaceutical company determine what are acceptable specifications for its product at the early stages of clinical development? Are there really enough test data at the start of the IND trials to "establish" specifications? As an industry, are we doing too many tests at the IND stage (i.e. following the "must do every test on someone's published list" syndrome), and then finding that specifications need to be set for each test we run? Are we feeling pressurised (either by regulatory agencies or by our own corporate cultures) to set unrealistic and tight specifications at the IND stage, and then later regretting how to justify loosening them when we gain more experience with the release testing or stability of our products? A discussion on how to set practical product specifications for biopharmaceutical products at the IND stage is presented. PMID- 9413683 TI - Characterization and establishment of specifications for biopharmaceuticals. AB - This paper describes the role of product characterization and product specifications in the context of an overall control strategy. Due to advances in analytical characterization and biotechnology manufacturing methods, product characterization can now be used as an effective means of assessing the impact of manufacturing process changes on product quality, safety and efficacy, hence obviating the need for repeating clinical testing each time a manufacturing process change is made. The ICH specifications guidance document can serve a key role in harmonising test requirements and regulatory practices with regard to produce characterization. Consumers, regulatory agencies, and biopharmaceutical manufacturers all stand to benefit from this harmonization process. PMID- 9413682 TI - Global regulatory considerations for unified assay specifications. AB - When developing a biotechnology product for global registration, there are several aspects to evaluate in an effort to unify specifications. These include differences between the United States, Europe and Japan, and Rest-of-World (ROW) countries with regard to the respective regulatory guidelines and pharmacopoeias in force, the state-of-the-art of product testing analytical methods, and the interval between submitting a registration dossier to different countries. In terms of regulatory guidelines, one country may have a monograph or required specifications for particular tests, for example the potency that a product has to meet before clinical trials can be initiated. For pharmacopoeias, different assay methods are required for sterility, general safety, and pyrogen testing, so that one may have to test a specific lot of a product at two or three different times to evaluate the same parameter, because of specific testing differences required for each country's pharmacopoeia. In addition, the state of analytical methods is always evolving and better analytical techniques become available. Sometimes, from starting with one set of tests, and based on the time in development, new tests may be added to the existing list of release specifications, because new analytical techniques have become available. Examining the global registration approval process for Betaseron, (interferon beta-1b) illustrates when specifications were able to be unified and when they were not. PMID- 9413684 TI - Raw material considerations. AB - Raw materials (RM) testing and control strategy form a major part of the foundation of a well-characterized protein (WCP) or biopharmaceutical. Raw materials may be present in the final vial as excipients or may have product contact earlier in processing. Manufacturers of WCPs should use a scientific approach to set acceptance criteria and test methods for bulk raw materials which are not present in dosage forms in substantial amounts. These methods and standards should enable the procurement of specified RMs which enable the reliable preparation of a product which consistently meets its quality attributes. Manufacturers of WCPs must use available pharmacopoeial standards for excipients and bulk process RMs which cannot be substantially removed during purification or other processing steps. Manufacturers should support efforts to harmonize pharmacopoeial standards for excipients. PMID- 9413685 TI - In-process testing and limits. PMID- 9413686 TI - The use of bioassays for the characterisation and control of biological therapeutic products produced by biotechnology. AB - The primary value of bioassays is that they alone directly assess biological activity of bioactive substances and products. Appropriately designed bioassays reflect the fundamental aspects of the biological activity of a bioactive molecule, including ligand-receptor binding, signal transduction processes (often poorly understood) and the final observed biological effects. Biological assays therefore complement physicochemical and biochemical procedures which normally only assess precise molecular structural features of complex molecules produced by biotechnology. Bioassays provide valuable information concerning the potency of biological products. This is essential for evaluating batch-to-batch consistency and stability. Bioassay data are crucial at all stages in the development of biological products, from early research work to final quality control of finished products. However, the type and design of bioassays may differ according to the information required and its intended use. The assays may or may not directly relate to the clinical use of the product. Bioassays can be difficult to perform and be time consuming, although this often reflects bad assay choice and/or design. Correct analysis of the assay results is essential if valid data are to be obtained. Standardisation, using correctly calibrated primary and secondary standards, is essential. In vivo bioassays are normally more unreliable than in vitro procedures, but in some cases the latter are either not available or do not address important biological characteristics of a product. Bioassays must be validated for their intended purpose and for the types of samples to be measured. Appropriate statistical analysis should be used to derive the significance and specifications of results. This needs to address both variability in samples and assay performance. Specifications (limits) for product acceptability need to be derived from real data using several batches of product. PMID- 9413688 TI - Recent developments in scanning laser ophthalmoscopy. PMID- 9413687 TI - The role of assay validation in specification development. AB - Three of the main components that contribute to the establishment of specifications relate to manufacturing consistency, clinical experience, and assay performance. The validation of an analytical control system for biotechnology pharmaceutical products represents a diverse analytical challenge. This paper focusses on the key components of assay validation and how they influence specification design. In particular, issues relating to precision, accuracy, and specificity are discussed in the context of their influence and impact on specification design. Specific examples are presented that encompass assays that are used for purity and potency and their interrelationships. PMID- 9413689 TI - Multifocal ERG recording by the VERIS technique and its clinical applications. PMID- 9413690 TI - Advances in the management of vitreomacular traction syndrome and macular hole. PMID- 9413691 TI - Use of autologous platelet concentrate in macular hole surgery: report of 77 cases. PMID- 9413692 TI - Autologous platelet concentrate in the surgical management of macular holes. PMID- 9413693 TI - Hemorrhagic macular cysts in Terson's syndrome and its implications for macular surgery. PMID- 9413694 TI - Complications in the removal of the posterior vitreous cortex. PMID- 9413695 TI - Epiretinal macular membranes: pathogenesis and treatment. PMID- 9413696 TI - Local therapy of CMV retinitis: a new therapeutical approach. PMID- 9413698 TI - Experimental therapies for age-related macular degeneration. PMID- 9413697 TI - A retained catheter for retrobulbar administration of interferon for age-related macular degeneration. PMID- 9413699 TI - Evaluation of macular therapy. PMID- 9413700 TI - Postinfarction use of beta-blockers in elderly patients. AB - beta-Adrenoceptor antagonists (beta-blockers) reduce mortality and recurrent myocardial infarction (MI) in older patients after both Q-wave MI and non-Q-wave MI. The effects of beta-blockers are to: (i) reduce complex ventricular arrhythmias, including ventricular tachycardia; (ii) increase the ventricular fibrillation threshold; (iii) reduce myocardial ischaemia; (iv) decrease sympathetic tone; (v) markedly attenuate the circadian variation of complex ventricular arrhythmias: (vi) abolish the circadian variation of myocardial ischaemia; and (vii) abolish the circadian variation of sudden cardiac death or MI. beta-Blockers reduce mortality in patients with MI and complex ventricular arrhythmias. In addition, they are excellent antianginal agents. Older persons with hypertension who have had an MI should be treated initially with a beta blocker. beta-Blockers reduce mortality in patients with: (i) diabetes mellitus who have had an MI; (ii) MI and congestive heart failure with an abnormal or normal left ventricular ejection fraction; and (iii) MI and an asymptomatic abnormal left ventricular ejection fraction. Severe congestive heart failure, severe peripheral arterial disease with threatening gangrene, greater than first degree atrioventricular block, hypotension, bradycardia, lung disease with bronchospasm, and bronchial asthma are contraindications to treatment with beta blockers. PMID- 9413703 TI - Adverse effects of thyroid hormones. AB - The adverse health effects of thyrotoxicosis have been carefully documented and most practitioners are familiar with the clinical consequences for the patient. Until recently, many patients experienced the adverse effects of excessive thyroxine dosages. Which can now be avoided by the application of highly sensitive immunometric assays for monitoring serum thyrotrophin (thyroid stimulating hormone; TSH) levels. However, sensitive monitoring of serum thyrotrophin levels has led to the frequent recognition of biochemical subclinical hyperthyroidism (isolated suppression of thyrotrophin). Because of the increased recognition of this condition, the adverse effects of thyroxine therapy can be divided into those associated with subclinical hyperthyroidism and those associated with the euthyroid state. Investigation of the potential clinical consequences of thyrotrophin-suppressing dosages of thyroxine has dominated studies over the last decade, with less attention being given to euthyroid patients. It appears that the adverse effects of thyroxine are considerably more common when serum thyrotrophin has been suppressed. They are usually manifested in older patients as increased bone mineral loss in postmenopausal women and as cardiac effects in patients with intrinsic heart disease. These patients may have subtle behavioural alterations and other clinically silent organ effects that occur infrequently. Children who are euthyroid while taking thyroxine occasionally develop pseudotumour cerebri shortly after starting hormone replacement for hypothyroidism. Otherwise, thyroxine dosages that render patients euthyroid, as evidenced by thyrotrophin values that are within the normal range, rarely cause adverse effects. Thus, avoidance of dosages that cause thyrotrophin suppression, when not clinically indicated, is the primary approach to the management of these adverse effects. PMID- 9413702 TI - Potential role of muscarinic agonists in Alzheimer's disease. AB - Alzheimer's disease (AD) is a common neurodegenerative disorder and a leading cause of death among the elderly. Recent advances in our understanding of the neurobiology of AD have provided scientific groundwork for the development of potentially more effective and less toxic treatment strategies for the disease. Some of the neuropathological hallmarks of AD include early and extensive degeneration of cortically projecting cholinergic neurons in the basal forebrain, and a reduced number of muscarinic acetylcholine receptors. Of note, neocortical muscarinic receptors of the M1 subtype are relatively preserved in the brains of patients with AD, whereas the presynaptic receptors, which are of the M2 subtype, are reduced in number. Therefore, activation of relatively intact postsynaptic mechanisms by muscarinic M1 receptor-specific agonists could theoretically be more efficacious in the treatment of AD compared with agents (e.g. acetylcholinesterase inhibitors) that predominantly act on dysfunctional presynaptic terminals. The administration of muscarinic agonists can demonstrably enhance cognition and significantly improve some of the disturbing behaviours in patients with AD. Recent advances in our knowledge of the molecular biology of muscarinic receptors, together with a better understanding of signal transduction pathways in AD, are likely to result in the development of receptor-specific muscarinic agonists that are more efficacious and less toxic. Moreover, preliminary evidence concerning the effects of muscarinic agonists on the processing of amyloid precursor protein and the formation of neurofibrillary tangles suggests that these agents might favourably alter the pathobiology of AD. PMID- 9413704 TI - Clinical pharmacokinetics of nifedipine. Implications for the care of the elderly. AB - Nifedipine, the prototype for the dihydropyridine class of calcium antagonists, has been available for 20 years and its efficacy as a vasodilator and an antihypertensive agent is well recognised. The development of the so-called nifedipine gastrointestinal therapeutic system (GITS), which allows once-daily administration, has modified and improved the overall therapeutic profile of nifedipine to such a significant extent that it might almost be considered a new drug entity. The nifedipine GITS is associated with distinct improvements in terms of patient compliance and convenience, and a reduced incidence of adverse effects. With regard to the care of the elderly, this 'new' drug offers the prospect of a well tolerated and effective treatment without major cost implications. PMID- 9413701 TI - Vitamin supplementation therapy in the elderly. AB - Vitamin supplementation in large dosages is increasingly common in the older population. Often, such supplementation is used in an attempt to improve an individual's health status. There have been claims that the effects of vitamins halt the normal aging process or prevent and cure disease. However, several recent studies have failed to demonstrate the efficacy of vitamin supplementation in preventing several types of cancer. In moderate dosages, supplementation with vitamin E (tocopherols) shows promise as a lipid antioxidant, and may reduce the risk of coronary heart disease. However, before vitamin E becomes an accepted medical therapy, further long term studies must be undertaken to examine the safety and efficacy of such therapy. An adequate intake of vitamins should be ensured by adherence to a well balanced diet. However, the elderly are prone to circumstances that may prevent them from eating a balanced diet. In addition, there are several age-related medical conditions that may predispose individuals to dietary and vitamin deficiencies. To prevent vitamin deficiency diseases and their associated morbidity, modest vitamin supplementation may be necessary. However, supplementation should be reserved for individuals with documented deficiency or who are at risk of developing such deficiencies, especially those who are homebound or institutionalised. Vitamins taken in large dosages should be considered as drugs. These medicines, which are obtainable over-the-counter, should be carefully regulated to prevent toxicity. PMID- 9413705 TI - Age-associated memory impairment. Normal aging or warning of dementia? AB - Aging causes deterioration in various aspects of memory performance in healthy adults. Different diagnostic classifications have been proposed for use in the characterisation of mild cognitive disorders associated with aging. One of the best established of these classifications is age-associated memory impairment (AAMI). Epidemiological data suggest that AAMI is a phenomenon of normal aging rather than a sign of progression from normal aging to a pathological state such as Alzheimer's disease. A number of studies that have combined neuropsychological, neuroradiological and neurophysiological data have provided evidence of distinct characteristics in individuals with AAMI. At present, however, AAMI does not appear to describe any homogeneous group of individuals. Moreover, the neuropsychological methods used to diagnose AAMI appear to be ambiguous. Thus, AAMI appears to occur in a highly heterogeneous group of individuals, and is of questionable clinical or theoretical significance. More reliable diagnostic approaches are needed for use in studies that are attempting to identify the risk factors for dementia or to find a treatment for very early dementia. PMID- 9413706 TI - Metrifonate. PMID- 9413707 TI - Metrifonate. A viewpoint. PMID- 9413708 TI - Metrifonate. A viewpoint. PMID- 9413709 TI - Gastro-oesophageal ulcers in man and horse: semblance and dissemblance. PMID- 9413710 TI - What really causes colic in horses? Epidemiology's role in elucidating the ultimate multi-factorial disease. PMID- 9413711 TI - Chronic colic in the mature horse: a retrospective review of 106 cases. AB - The clinical features of 106 horses presenting with chronic colic examined over a 5 year period were reviewed. Chronic colic was defined as colic signs observed daily for 3 days or longer, except when masked by analgesics. The diagnosed causes of chronic colic included colonic impaction (31%), peritonitis (16%), enteritis/colitis (7%), colonic displacement/torsion (6%) and lymphosarcoma (4%). A variety of other diseases were diagnosed in a small number of cases, including intestinal adhesions, ileal obstructions, grass sickness, liver disease, caecal impactions, thromboembolic disease, intussusceptions etc. No diagnosis was reached in 8% of cases. Diagnosis was relatively easily achieved in most cases of colonic impaction and peritonitis by results of transrectal palpation and evaluation of peritoneal fluid. Specific clinical features, transrectal palpation, abdominal paracentesis and laboratory evaluations of blood were helpful in the diagnosis of some of the other diseases, but exploratory laparotomy remained necessary in a few cases to achieve a diagnosis. PMID- 9413712 TI - Recurrent colic in the mature horse: a retrospective review of 58 cases. AB - The clinical features of 58 horses presenting with recurrent colic examined over a 5 year period were reviewed. The horses were categorised into 3 groups on the basis of the history of colic episodes. Recurrent transient colic Group 1 was characterised by 3 or more episodes of transient colic (of apparently similar type) occurring within one month. Recurrent transient colic Group 2 was characterised by 3 or more episodes of transient colic occurring within one year. Recurrent prolonged colic was characterised by 3 or more episodes of prolonged colic occurring within one year. Fifteen horses were classified as recurrent transient colic Group 1. This group had the highest mortality rate 53%. They included 3 horses with lymphosarcoma, 2 with intussusceptions, 2 with thromboembolic disease/verminous arteritis and 2 with partial ileal obstructions. The recurrent transient colic Group 2 comprised 27 cases with a mortality rate of 4%. Nine of these horses were affected by spasmodic colic. The recurrent prolonged group involved 16 horses with a mortality rate of 31%. These included 3 cases of recurrent colonic impaction and 2 cases each of lymphosarcoma, 2 thromboembolic disease, 2 partial ileal obstructions and 2 intestinal adhesions. PMID- 9413713 TI - Effects of omeprazole on healing of naturally-occurring gastric ulcers in thoroughbred racehorses. AB - Seventeen Thoroughbred horses with moderate to severe gastric ulceration were purchased from a race track within 10 days of racing and were treated once daily with either omeprazole (9 horses) or vehicle (8 horses) and evaluated gastroscopically for ulcer healing. Horses were administered omeprazole (1.5 mg/kg bwt/day) or vehicle by nasogastric tube once daily. Gastroscopic examination was performed on Days 0, 4, 7, 11, 14, 17, 21, 24 and 28, until lesions healed completely. Selected images of gastric lesions were captured by computer at each endoscopic examination, with a measuring caliper included in captured images. The area and perimeter of lesions were measured by computer and healing rates of specific lesions were determined by calculating the rate of linear advance of the margins toward the centre of the lesion. Additionally, the number of days to complete healing of the entire gastric squamous mucosa was compared between treatment groups. Gastric lesions healed at a significantly faster rate in horses receiving omeprazole than in vehicle-treated horses (P < 0.001). Complete healing of the entire stomach occurred in 10-21 days in omeprazole-treated horses, and 14-28 days in 3 of 8 vehicle-treated horses, with the remaining vehicle-treated horses having unhealed lesions on Day 28. In addition, 5 vehicle-treated horses developed new lesions in the squamous epithelial mucosa during the trial; no new lesions were observed in the omeprazole-treated group. PMID- 9413714 TI - A scoring system for gastric ulcers in the horse. AB - Five investigators familiar with gastric ulcer disease in horses met to establish a scoring system that could be utilised in future studies. Slides of gastric lesions were viewed and discussed and a scoring system established that required the nonglandular and glandular portions of the stomach to be graded separately. Each portion of the stomach (glandular and nonglandular) received a score for number of ulcers present and a score for severity of ulcers which resulted in each stomach receiving 4 separate scores. After the grading system was developed, each investigator independently graded 16 horses with gastric ulcer disease that had been previously recorded on video tape. The results of each investigator's scores were then compared. There was a variability between observers in the scores for severity of both nonglandular and glandular lesions but the variability was not significant. The variability between observers for the number of glandular lesions was also not significant. This implied that there was consistency between the 5 observers in the way severity of lesions was scored and the number of glandular lesions. However, there was a significant variability between observers for the number of nonglandular lesions which implied agreement on this observation was more variable. PMID- 9413715 TI - Expression of transforming growth factor-beta 1 in normal and dyschondroplastic articular growth cartilage of the young horse. AB - This study describes the distribution pattern of transforming growth factor-beta 1 (TGF-beta 1) mRNA and protein in normal pre- and post natal growth cartilage and alterations present in lesions of dyschondroplasia (osteochondrosis). TGF beta 1 expression and immunoreactivity have been investigated by in situ hybridisation and immunolocalisation in the articular/epiphyseal growth cartilage of the lateral trochlear ridge of the distal femur. Cartilage was obtained from 19 normal Thoroughbred horses (5 prenatal and 14 post natal horses) and 15 post natal horses with dyschondroplasia (DCP). TGF-beta 1 mRNA expression and immunoreactivity were detected in the proliferative and upper hypertrophic zones in both pre- and post natal normal articular/epiphyseal cartilage. However, mRNA itself was only detected in the mid- and lower hypertrophic zones. Immunoreactivity was identified intracellularly with some nuclear staining observed. In focal lesions of DCP mRNA expression and immunoreactivity were reduced compared to normal cartilage, but strong mRNA expression was observed in the chondrocyte clusters immediately surrounding a lesion of DCP. The results described in this study demonstrate alterations in TGF-beta 1 dyschondroplastic lesions and indicate that it could be involved in the pathogenesis of this condition in the horse. PMID- 9413716 TI - Effects of insulin and insulin-like growth factors I and II on the growth of equine fetal and neonatal chondrocytes. AB - The effects of insulin and insulin-like growth factors (IGFs) I and II on fetal and foal chondrocytes were investigated in vitro. Chondrocytes from the lateral trochlear ridge of the distal femur were obtained from 2 fetuses (280 and 320 days gestation) and one 4-day-old foal and cultured. Membrane proteins consistent with type 1 and type 2 IGF receptors were demonstrated by radioligand cross linking and equilibrium binding analysis. It was demonstrated that both IGF-I and IGF-II acted as mitogens for isolated equine chondrocytes when present as the sole mitogenic factor in monolayer culture. It was further shown that whereas insulin was able to promote the survival and expansion of cell populations of chondrocytes in culture there was significantly reduced mitogenic stimulation compared to the IGFs. These results suggest that the role of insulin in growth cartilage may be to promote chondrocyte survival, or to suppress differentiation/apoptosis. This supports the hypothesis that relative hyperinsulinaemia may be a contributory factor to equine dyschondroplasia (osteochondrosis). Understanding of contributory, and possibly triggering factors such as this may allow the development of modified methods of husbandry which minimise the risk of disease in populations with a known predisposition. PMID- 9413717 TI - Prospective study of equine colic incidence and mortality. AB - A prospective study of one year was conducted on 31 horse farms to obtain population based estimates of incidence, morbidity and mortality rates of equine colic. Farms with greater than 20 horses were enrolled by randomly selecting horse owners from 2 adjacent counties of Virginia and Maryland. Descriptive information for 1427 horses was collected at the initiation of the study and updated at 3 month intervals. Time on the farm during the study was tabulated for each horse. When colic was reported by the owner, investigators visited the farm to obtain information about the colic. The crude incidence density rate of colic was 10.6 colic cases/100 horse-years, based on 104 cases/358,991 horse-days. The median farm specific incidence density rate was 7 cases/100 horse-years, and the range for individual farms varied from 0 to 30 colic cases/100 horse-years. A specific diagnosis was not made for 84 (81%) of colic episodes. Seventy colic episodes (67%) were treated by a veterinarian. Drugs were used in 83 (80%) colic episodes, and 78 (75%) of colic cases were mild, requiring no treatment or resolving after only one treatment. Four horses required colic surgery. Fourteen (13%) horses had more than one episode of colic during the year. Mortality from all causes of death was 2.5 deaths/100 horse-years, mortality rate for colic was 0.7 deaths/100 horse-years. Proportional mortality rate of colic, 28%, was higher than for any other cause of death. Horses less than age 2 years or greater than age 10 years had lower incidence than horses age 2-10 years. No difference in colic risk between genders was identified. Arabian horses had the lowest and Thoroughbreds the highest breed specific incidence rates. Horses used for eventing, or in training had a statistically significant higher incidence rate of colic compared to mature horses with no use (pets, retired, on pasture with no stated purpose). Horses used for lessons or with no use had the lowest incidence rates. PMID- 9413718 TI - Prospective study of equine colic risk factors. AB - A 1 year prospective study was conducted on 31 horse farms to identify risk factors for equine colic. Farms were randomly selected from a list from 2 adjacent counties of Virginia and Maryland, USA. The association between colic and farm or individual horse risk factors related to management, housing, pasture, use, nutrition, health and events was first examined by univariate statistical analysis. Individually significant (P < = 0.25 for farm factors, P < = 0.10 for horse factors) variables were used in a stepwise multivariable forward logistic regression to select explanatory factors (P < = 0.05). Analysis was conducted at 2 levels: farm and individual horse with farm specified as a random effects variable. No farm-level variables were significant. Significant horse level variables included: age, odds ratio (OR) = 2.8 for horses age 2-10 years compared to < 2 years; history of previous colic, OR = 3.6 relative to no colic; changes in concentrate feeding during the year (1 per year, OR = 3.6, more than 1, OR = 2.2) relative to no changes; more than 1 change in hay feeding during the year, OR = 2.1 relative to no changes; feeding high levels of concentrate (> 2.5 kg/day dry matter, OR = 4.8, > 5 kg/day dry matter, OR = 6.3) relative to feeding no concentrate; and vaccination with monocytic ehrlichiosis vaccine during the study, OR = 2.0 relative to no vaccination. Feeding a whole grain with or without other concentrate components reduced risk, OR = 0.4, relative to feeding no whole grain. Results of the study suggest that diet and changes in diet are important risks for colic in a population of horses on farms. PMID- 9413719 TI - Endoscopic examination of the carpal flexor tendon sheath in horses. AB - This study was undertaken to design a safe technique to examine the carpal flexor tendon sheath (carpal sheath) of horses endoscopically, using an arthroscope. The limbs from 15 horses were used to study the normal anatomy of the carpal sheath and related structures, establish a safe approach and endoscopic technique, and determine the normal endoscopic appearance of the sheath. Major arteries, veins and nerves, present within and around the sheath, left few 'safe' areas to insert the endoscope. Several portals were assessed and a distal lateral approach was found to be safest and to allow adequate visualisation of most of the sheath. The surgical technique and normal endoscopic findings are described in detail and discussed. PMID- 9413720 TI - The effect of age and diet on the oral glucose tolerance test in ponies. AB - To evaluate the effects of age and diet on the oral glucose tolerance test (OGTT) in healthy ponies, OGTTs were performed on 2 groups of British native breed ponies (Group A: 7 foals [6-9 months], Group B: 7 mature individuals [6-13 years]) when maintained on either a high fibre pelleted ration only (Groups A and B) or a hay only diet (Group B). Plasma glucose response, following oral glucose administration, for Group A (basal plasma glucose concentration [Glu0] 4.6 +/- 0.4 mmol/l (mean +/- s.d.) increasing to 11.5 +/- 1.3 mmol/l at 90 min) was significantly different (P < 0.05) from that observed for Group B (Glu0 of 4.3 +/ 0.2 mmol/l increasing to 6.8 +/- 1.3 mmol/l at 90 min), when fed the same diet. For Group B ponies, the plasma glucose response, following oral glucose administration, was significantly different (P < 0.05) when fed hay only (Glu0 4.6 +/- 0.4 mmol/l increasing to 9.6 +/- 2.1 mmol/l at 150 min) compared to when fed the high fibre pelleted ration. These results indicate that both age and diet have a significant effect on plasma glucose concentrations measured during an OGTT. PMID- 9413721 TI - Technical validation of a face mask adapted for dry powder inhalation in the equine species. AB - Development of dry powder inhalation (DPI) for horses requires the use of an adapted face mask. In experiment I, 4 masks (A, B, C and D) were tested and factors influencing the delivery of the dry powder were determined. Mask A was one which is commercially available for metered-dose inhalation. Mask B had the same shape as Mask A but an airtight rubber seal was added for the connection between the mask and horse's head. Mask C was a prototype adapted for DPI with connection for the DPI device between the nostrils, airtight expiratory valves in front of each nostril and airtight rubber seal to attach the mask on the horse's head. Mask D was the same as Mask C but the airtight expiratory valve was situated in front of one nostril and the connection for the DPI device was placed in front of the other nostril. Inhalet emptying and peak inspiratory pressure were measured on 5 healthy horses with each face mask. Both Masks A and B gave a low rate of inhalet emptying. Inspiratory pressures created in Masks C and D were negative enough to ensure inhalet emptying rates of mean +/- s.d. 98.28 +/- 1.79% and 100% respectively. In experiment 2, the face masks giving the greatest inhalet emptying were used to test the therapeutic efficacy of ipratropium bromide DPI. This was tested on 6 horses suffering from acute exacerbation of chronic obstructive pulmonary disease (COPD). At a dose of 200 micrograms/100 kg bwt, ipratropium administered with Mask D improved significantly pulmonary function measurements compared to baseline values and placebo inhalation. With Mask C, a double dose of ipratropium (400 micrograms/100 kg bwt) was necessary to improve these parameters compared to baseline values. This indicated the importance of locating the DPI device in front of one nostril. It was concluded that inhalet emptying is correlated to inspiratory pressures measured in the face masks. Secondly, these pressures are in turn dependent on the air-tightness of the mask, i.e. air-tightness of the expiratory valve and close connection between the horse's head and the mask. Thirdly, position of the DPI device in front of a nostril allows bronchodilation at a dose half that required when the device is placed between the nostrils. Finally DPI using Mask D (EquiPoudre) is a rapid, effective and well tolerated inhalation treatment for COPD horses. PMID- 9413722 TI - Surgical treatment of subchondral cystic lesions of the third metacarpal bone: results in 15 horses (1986-1994). AB - Subchondral cystic lesions (SCLs) in the condyle of the third metacarpal bone (MCIII) were surgically treated in 15 horses. The median age at presentation was 18 months (range 10 months-12 years) with 10 of 15 horses less than age 2 years. The SCLs were confined to the front limbs in all cases with 2 horses having bilateral lesions. Lesions were isolated to the medial condyle(s) of MCIII in 13 of 15 horses; a cystic lesion occurred in the lateral condyle in one horse and in the sagittal ridge in one horse. One horse with bilateral lesions had an additional cystic lesion located in the right medial femoral condyle. Fourteen of 15 horses had a history of moderate lameness attributable to the metacarpophalangeal joint; the lesion was an incidental finding in one horse. Duration of lameness ranged from 4 weeks to 8 months and was either acute in onset, or occurred intermittently and was associated with exercise. Fetlock flexion significantly exacerbated the lameness in all cases. Synovial effusion was absent in 8 (53%) cases. Cystic lesions were curetted arthroscopically in 12 horses, and through a dorsal pouch arthrotomy in 3 horses. Concurrent osteostixis of the cystic cavity was performed in 7 horses. Two horses were treated arthroscopically for osteochondral fragmentation of the proximodorsal aspect of the proximal phalanx one year following surgical curettage of the SCL. Twelve of 15 horses (80%) were sound for intended use following surgical treatment. Two horses did not regain soundness and follow-up information was unavailable for one horse. Total period of follow-up was 1-6 years. Follow-up radiographic examinations were available for 9 horses. Mild periarticular osteophyte formation and enthesiophyte formation at the dorsal joint capsular attachments was present in 5 of the 9 horses. Bony ingrowth of the cystic lesion was detectable in 8 horses and enlargement of the cystic cavity was observed in one horse. Based on the information gained from this study, it would appear that surgical treatment of SCLs in the distal metacarpus can result in a favourable outcome for athletic use. PMID- 9413723 TI - Stress response to chronic inflammation in the horse. AB - Five clinically healthy Thoroughbred geldings were injected with Freund's adjuvant 3 times to induce a chronic inflammatory response. Blood was collected at various times before and after adjuvant administration. Clinical responses (rectal temperature and general demeanor) were also monitored. Adjuvant injection induced increases in rectal temperature and plasma fibrinogen concentration (maximum levels measured were mean +/- s.d. 39.7 +/- 0.5 degrees C and 8.2 +/- 0.3 g/l, respectively), indicative of an inflammatory response. A mild clinical depression was also observed in the horses for 24 h after the first injection of adjuvant only. Plasma cortisol levels decreased significantly from control levels of mean +/- s.d. 187.7 +/- 24.3 nmol/l to a minimum of 80.2 +/- 22.1 nmol/l (P < 0.01) 9 days after the first injection of adjuvant. Conversely, plasma insulin levels increased after the first injection of adjuvant to a maximum (96.7 +/- 15.2 iu/ml; P < 0.01) 12 days later, while plasma glucose concentrations tended to decline. A control group of horses to rule out contemporary environmental influences on the physiological and biochemical indices measured was not included in this study. The results show that chronic inflammation in the horse depressed resting plasma cortisol concentrations. PMID- 9413724 TI - Microtubular defect in equine spermatozoa associated with infertility. PMID- 9413725 TI - Comparison of indirect immunofluorescence for Ehrlichia phagocytophila and Ehrlichia equi in horses. PMID- 9413726 TI - Thoracic vertebral malformation in two horses. PMID- 9413727 TI - Anion fluxes, volume regulation and electrical activity in the mammalian pancreatic beta-cell. PMID- 9413728 TI - Modulation of Ca2+ and K+ permeabilities by oxotremorine-m (Oxo-m) in rodent pancreatic B-cells. AB - The effects of the muscarinic agonist oxotremorine-m (Oxo-m) on 45Ca and 86Rb fluxes, insulin secretion, cytoplasmic Ca2+ concentration ([Ca2+]i) and membrane potential in pancreatic B-cells were studied. Oxo-m (40-200 microM) increased the [Ca2+]i by about 250 nM, irrespective of the glucose concentration present in the medium (2.8-22 mM). This effect was reduced by 50% upon the addition of EGTA. Oxo m (50 microM) increased the 45Ca efflux from islets perifused in the absence or presence of [Ca2+]o, although under the former condition this efflux was transient. The difference between effluxes measured in the absence and presence of [Ca2+]o represents the sustained second component, which presumably reflects Ca2+ influx. In both the absence and presence of 11.2 mM glucose. Oxo-m (50 microM) transiently increased 86Rb efflux. In the presence of glucose, Oxo-m provoked a transient polarization of the B-cell membrane associated with an increase in the K+ permeability values. K+ permeability returned to basal values (no Oxo-m) after 1-2 min. These results indicate that the initial phase of Oxo-m induced insulin secretion depends partially on intracellular Ca2+ release, and that the sustained enhancement of release depends on Ca2+ influx. The participation of a calcium release-activated current (ICRAC) is proposed to explain the sustained small changes in membrane potential. PMID- 9413729 TI - The enzymic potential of tissue kallikrein (rK1) in rat submandibular saliva depends on whether it was secreted via constitutive or regulated pathways. AB - The enzymic activity and immunoreactivity of rat tissue kallikrein (rK1) secreted at rest by granular duct cells of unstimulated submandibular glands has been compared with that secreted on autonomic nerve stimulation. Although a direct vesicular, constitutive secretory pathway operates for rK1 secretion from granular duct cells of unstimulated and parasympathetically stimulated glands the rK1 was not present in a pro-form and actually showed a greater enzymic activity per unit immunoreactive protein than the granule-derived rK1 in sympathetically evoked saliva. Constitutively secreted rK1 was found to be in a single chain molecular form by reducing SDS gel electrophoresis. In contrast rK1 secreted from the storage granule pool of granular duct cells on sympathetic nerve stimulation was present in much higher amounts and occurred in both one-chain and two-chain forms as revealed by SDS gel electrophoresis under reducing conditions. The lower enzymic potential of rK1 in sympathetically evoked saliva might be accounted for by its conversion to a two-chain form. PMID- 9413730 TI - Increased sensitization of the myofilaments in rat neonatal portal vein: a potential mechanism. AB - The contractile regulation of neonatal smooth muscle was studied in rat neonatal portal vein. Strips of beta-escin-permeabilized portal vein from 3- to 5-day-old rats and 5- to 6-week-old rats were mounted on a 'bubble chamber' in calcium solutions buffered with 10 mM EGTA. Although the overall tension development was lower in neonatal muscle as expected, the calcium sensitivity of the neonatal permeabilized portal vein was significantly higher than in developed portal vein. Endothelin-1-induced sensitization of the myofilaments was investigated. Endothelin-1 produced an increase in tension of permeabilized neonatal portal vein in a calcium solution buffered with 10 mM EGTA. This sensitization was proportionally higher in neonatal than in developed smooth muscle, despite similar initial submaximal calcium contractions. GTP gamma S-induced calcium sensitization was also proportionally higher in neonatal permeabilized strips than in fully developed smooth muscle. These changes may be due to alterations in the intracellular signalling pathways which mediate calcium sensitization in smooth muscle. As some endothelin-1-mediated responses are known to occur via activation of the heterotrimeric GTP-binding protein, Gq, the levels of protein expression of Gq alpha were studied. In membrane preparations from neonatal rat portal vein the expression of Gq alpha was significantly higher than in portal vein membrane preparations loaded with equal protein from 5- to 6-week-old rats. In conclusion, agonist-induced sensitization of the myofilaments was higher in neonatal rat portal vein than in fully developed portal vein. This difference is the result of changes in intracellular signalling and may be partly produced by the greater expression of Gq alpha observed in neonatal portal vein. PMID- 9413731 TI - Reflex vascular responses to alterations in abdominal arterial pressure and flow in anaesthetized dogs. AB - The existence of abdominal arterial baroreceptors has long been controversial. Previously difficulties have been encountered in localizing a stimulus to abdominal arteries without affecting reflexogenic areas elsewhere. In these experiments, using anaesthetized dogs, the abdomen was vascularly isolated at the level of the diaphragm, perfused through the aorta, and drained from the inferior vena cava to a reservoir. Changes in abdominal arterial pressure were effected by changing the perfusion pump speed. During this procedure the flow back to the animal from the venous outflow reservoir was held constant. Increases and decreases in abdominal arterial pressure resulted, respectively, in decreases and increases in perfusion pressure to a vascularly isolated hind-limb and in some dogs also a forelimb. Responses were significantly larger when carotid sinus pressure was high (120-180 mmHg) than when it was low (60 mmHg). Responses were still obtained after cutting vagus, phrenic and splanchnic nerves, but were abolished by spinal cord lesion at T12. These experiments provide evidence for the existence of abdominal arterial baroreceptors. The afferent pathway for the reflex vasodilatation appears to run in the spinal cord. PMID- 9413732 TI - The renal response of sheep to intraportal infusion of glucagon. AB - The effects on renal functions of infusing glucagon (100 ng kg-1 min-1) into the portal vein system were studied in young anaesthetized sheep kept on a diet with normal protein and digestible energy contents. The clearance measurements of the left kidney functions showed a decreased urine flow rate (from 0.36 +/- 0.04 to 0.18 +/- 0.03 ml min-1; P < 0.01) within the first 10 min of glucagon infusion, falling to 0.07 +/- 0.02 ml min-1 (P < 0.01) during the next 10 min. The glomerular filtration rate also decreased from 17.56 +/- 2.92 ml min-1 to its lowest value, 3.34 +/- 1.02 ml min-1, (P < 0.01) after 20 min of glucagon infusion. Decreases in urea excretion (from 53.1 +/- 6.46 to 3.82 +/- 1.27 mumol min-1; P < 0.01) and osmotic clearance (from 0.67 +/- 0.13 to 0.09 +/- 0.02 ml min-1; P < 0.01) occurred during the first 20 min. The plasma level of glucose increased from 4.01 +/- 0.53 to 8.33 +/- 1.12 mmol l-1 (P < 0.05) 20 min after the start of glucagon infusion. Twenty minutes after the start of glucagon infusion, the values of all the parameters measured began to recover, although they did not reach the control level, even after 1 h. Blood pressure was stable during the whole experiment. The results show that the effects of intraportally infused glucagon on kidney functions in sheep are the opposite of those obtained from similar experiments in laboratory animals with simple stomachs or in humans. PMID- 9413733 TI - Adipose tissue development during early postnatal life in ewe-reared lambs. AB - This study examines the precise time course that brown adipose tissue (BAT) takes to adopt the characteristics of white adipose tissue in postnatal lambs. Perirenal adipose tissue was sampled from ewe-reared lambs within 1 h of birth and at 1, 2, 4, 7, 14, 21 and 30 days of age and analysed for the amount of mRNA for uncoupling protein (UCP), the amount and activity of UCP, and protein, mitochondrial protein and lipid content. This was combined with measurements of colonic temperature and jugular venous plasma concentrations of thyroid hormones and insulin-like growth factor-1 (IGF-1). Over the first 4-7 days of age, large quantities of UCP mRNA were associated with a peak in plasma triiodothyronine concentration at 2 days of age followed by a maximal amount and activity of UCP at 4 days and a basal colonic temperature of 39.3 degrees C. Between 7 and 30 days there was a large increase in lipid deposition as the amount and activity of UCP and the amount of UCP mRNA declined to basal values and colonic temperature was maintained at 40 degrees C. A significant positive relationship between perirenal adipose tissue lipid content and plasma IGF-1 concentration was observed throughout the study period. It is concluded that ovine adipose tissue maturation occurs in two distinct phases over the first month of life. The precise time scale of this process could be regulated in part by the lamb's body temperature which determines whether adipose tissue is required for heat production (i.e. BAT) or as an endogenous energy source (i.e. white adipose tissue). PMID- 9413734 TI - Influence of feeding and ambient temperature on thermoregulation in newborn lambs. AB - This study examined the effect of ambient temperature and feeding on brown adipose tissue (BAT) function and thermoregulation in lambs born either vaginally at term or by Caesarean section close to term. Immediately after birth lambs were placed in a warm (30 degrees C) or cool (15 degrees C) ambient temperature and measurements of colonic temperature and heat production recorded for 6 h. Lambs were fed 50 ml of colostrum when 5 h old. The amount of uncoupling protein and level of guanosine 5'diphosphate (GDP) binding in BAT was higher in vaginally delivered lambs than in lambs delivered by Caesarean section. For each delivery group, GDP binding was greater in lambs maintained at 30 degrees C than in lambs maintained at 15 degrees C. O2 consumption, CO2 production and colonic temperature only increased after feeding in lambs born by Caesarean section and maintained at 30 degrees C, a response that was accompanied by a decreased incidence of shivering. Irrespective of delivery temperature, plasma thyroid hormone concentrations and noradrenaline content of BAT were lower in lambs born by Caesarean section than in those born vaginally. Plasma cortisol concentrations were higher in lambs delivered by Caesarean section, as was adrenaline content of BAT in these lambs maintained at 30 degrees C. It is concluded that the thermoregulatory response to feeding in terms of changes in both recruitment of shivering and colonic temperature were observed only in lambs delivered by Caesarean section. PMID- 9413735 TI - The effect of sodium citrate ingestion on the metabolic response to intense exercise following diet manipulation in man. AB - Feeding a high-carbohydrate (CHO) diet and administration of alkalinizing agents have both been shown to improve performance in high-intensity exercise. The effect of these treatments in combination was investigated in the present study. Six healthy male subjects exercised to exhaustion on an electrically braked cycle ergometer at a power output equivalent to 100% of their maximum oxygen uptake (VO2,max) on four separate occasions. Each subject consumed either a diet with the same composition as his normal diet (termed the experimental normal (N) diet; 54 +/- 7% CHO, 13 +/- 2% protein, 33 +/- 7% fat) or a high-CHO diet (81 +/- 2% CHO, 13 +/- 2% protein, 6 +/- 1% fat) that had the same energy and protein content for the 3 days prior to the exercise tests. Subjects then ingested either a placebo (CaCO3) or trisodium citrate (0.3 g (kg body mass)-1) 3 h before exercise. Time to fatigue was not different between experimental conditions. Consumption of the high-CHO diet had no effect on blood acid-base status, but the ingestion of sodium citrate induced a mild metabolic alkalosis after both the N diet and the high-CHO diet. This alkalinizing effect was also evident after exercise, since blood pH, plasma bicarbonate and blood base excess were higher (P < 0.05) after the ingestion of sodium citrate than under the placebo conditions. The changes in blood lactate, pyruvate and glucose and plasma glycerol after exercise were similar for all experimental conditions. Blood lactate, glucose and pyruvate and plasma glycerol concentrations increased from resting values (P < 0.01) following exercise but this increase was similar under all experimental conditions. These data demonstrate that when the energy and protein content of the diets is the same, exercise capacity and the metabolic response to intense exercise are similar following consumption either of a high-CHO diet or a more normal diet. Acute ingestion of sodium citrate prior to exercise resulted in a reduction in post-exercise acidosis despite a blood lactate concentration that was similar to that observed after the ingestion of a placebo, but did not affect exercise performance under the conditions of this study. PMID- 9413736 TI - The effects of buspirone on perceived exertion and time to fatigue in man. AB - Male subjects exercised at 80% maximal rate of O2 uptake (VO2,max) following oral administration of either placebo or the partial 5-HT1A agonist buspirone (45 mg), using a paired design. Ratings of perceived exertion were higher following buspirone and time to volitional fatigue (median and inter-quartile range) fell significantly by approximately a third from 26 min (24-30 min) on placebo to 16 min (11-19 min) following buspirone. Serum prolactin was significantly elevated following buspirone administration, indicating increased hypothalamic 5-HT1A receptor stimulation. There were no significant differences in blood lactate or serum glucose between the trials. This study supports the possible central modulation of exercise tolerance by serotonergic pathways, although a role for dopamine cannot be excluded. PMID- 9413737 TI - Immunohistochemical localization of type II and type I collagens in articular cartilage of the femoral head of dexamethasone-treated rats. AB - The immunohistochemical localization of type II and type I collagens was examined in the articular cartilage of the femoral head of growing rats injected systemically with 5 mg kg-1 dexamethasone for 2 weeks every other day. The intensities of immunostaining for type II collagen, measured by microphotometry, was highest in the flattened cell layer and high in the hypertrophic cell layer, moderate in the proliferative cell and transitional cell layers and low in the superficial layer. After dexamethasone administration, the intensities decreased markedly in the flattened cell layer and slightly in the hypertrophic cell layer, although the decreases in other layers were negligible. The staining intensities for type I collagen were highest in the flattened cell layer, low in the superficial and transitional cell layers and very low in the proliferative and hypertrophic cell layers. After dexamethasone administration, the intensities increased markedly in the flattened cell layer and slightly in the superficial and proliferative cell layers, but did not change in the transitional and hypertrophic cell layers. Thus, dexamethasone administration caused a decrease in type II collagen and an increase in type I collagen in the matrix of the surface portion of articular cartilage. The composition of isoforms of collagen in the matrix changed after the steroid administration. The results strongly that the shift in collagen composition from type II to type I predominance is a cause of the degeneration of the articular cartilage after glucocorticoid administration. PMID- 9413738 TI - An immunohistochemical and morphometric study on astrocytes and microvasculature in the human cerebral cortex. AB - In this study, astrocytes and microvessels of the human cerebral cortex were analysed morphometrically with the aim of acquiring quantitative information on the glio-vascular relationships, considered to be of great importance in the formation and functioning of the blood-brain barrier. Immunohistochemistry for the astrocytic marker, glial fibrillary acidic protein, was used with a computerized image analysis system. The brain tissue was embedded using the progressive lowering of temperature method, and the image analyser was applied to semithin sections subjected to immunogold-silver staining and viewed by epipolarization microscopy. The results show that, in the human cerebral cortex, astrocytes cover 11.4% of the cortex area and that their perivascular processes are nearly as extensive as the vascular bed (0.8% versus 1.72% of the cortex area). These processes form a virtually continuous sheath around the vascular walls, only 11% of the vessel perimeter lacking this astrocytic glia covering. The present results, compared with previous unpublished data obtained by conventional immunocytochemical procedures on wax sections, indicate that low temperature methods combined with gold-silver immunolabelling on semithin sections significantly improve the detection of immunoreactivity and the performance of the image analyser. PMID- 9413740 TI - Nitric oxide synthase immunoreactive nerves in rat and ferret salivary glands, and effects of denervation. AB - Nitric oxide has been implicated in mechanisms mediating nerve-evoked vasodilatory and secretory responses in salivary glands. In the present study, the occurrence and distribution of nitric oxide synthase (NOS)-immunoreactive nerves in ferret and rat salivary glands were investigated using immunocytochemistry with rabbit and sheep NOS antisera, and using NADPH diaphorase enzyme histochemistry. In the parotid, submandibular and sublingual glands of the rat and the ferret, NOS-immunoreactive varicose terminals encircled acini and arteries of various sizes. In the ferret, collecting ducts were also supplied with NOS-immunoreactive fibres. In the rat, only the granular ducts of the submandibular gland were supplied with such fibres. The NOS-immunoreactive innervation of acinar cells was more abundant in the rat than in the ferret, whereas the opposite was true for the innervation of blood vessels. No NOS immunoreactivity was observed in the vascular endothelium. In both species, NOS positive ganglionic cell bodies were found in the hilar regions of the submandibular and sublingual glands, whereas none could be detected in the parotid glands. NADPH-diaphorase reactivity had the same neuronal distribution as NOS immunoreactivity and, in addition, NADPH-diaphorase reactivity was expressed in ductal epithelium. Neither sympathetic denervation (by removal of the superior cervical ganglion) nor treatment with the sensory neurotoxin capsaicin reduced the NOS-immunoreactive innervation of the parotid gland. However, parasympathetic denervation (by cutting the auriculo-temporal nerve) caused an almost total disappearance of the NOS-immunoreactive innervation. The present findings provide a morphological background to the suggested role of nitric oxide in parasympathetic secretory and vascular responses of salivary glands. PMID- 9413739 TI - Immunohistochemical detection of nucleolar protein p120 in paraffin-embedded tissues. AB - The monoclonal antibody FB-2 recognizes the antigen p120-kDa protein (p120), associated with the nucleolar matrix. p120 has originally been reported as expressed and detectable in malignant and non-neoplastic proliferating cells, but not in most normal resting tissues and benign tumours. In the present study, a reliable immunostaining method was used to detect p120 on formalin-fixed, paraffin wax-embedded tissue, testing it on 148 samples from different neoplastic and non-neoplastic tissues from different organs (breast, colon, lung, prostate, bladder, lymph nodes, skin, tongue and liver). The immunostaining was performed after the application of a specific antigen-unmasking protocol based on six consecutive cycles of microwave oven heating. Under these retrieval conditions, p120 antigen was clearly detectable, not only in hyperplastic and malignant cells, but also in stromal and normal non-proliferating cells of all the tissues evaluated. Our results show that the nucleolar protein p120 can be detected by routine immunohistochemistry in formalin-fixed, paraffin-embedded tissue and is expressed in all nucleated cells under any biological condition. PMID- 9413741 TI - Lectin histochemical HPA-binding pattern of human breast and colon cancers is associated with metastases formation in severe combined immunodeficient mice. AB - Metastasis formation is a major clinical problem in cancer treatment, and no significant progress in the treatment of metastatic spread has been made. This apparent lack of progress is partly caused by the absence of clinically relevant animal models of metastases. The binding of the lectin Helix pomatia agglutinin (HPA) has been associated with a poor prognosis in breast and colon cancer patients. HPA-positive and -negative human breast and colon cancer cell lines were transplanted into severe combined immunodeficient (SCID) mice. HPA-positive breast cancer cell lines (MCF-7 and T47D) metastasized in SCID mice, whereas the HPA-negative ones (BT20, HS578T and HBL100) did not. The HPA-positive colon cancer cell line HT29 metastasized, while the HPA-negative ones (COLO320DM, SW480 and SW620) did not. However, in two of eight SCID mice inoculated with the HPA negative colon cancer cell line, CACO2 metastatic deposits were found. Despite this exception, HPA binding is a good indicator of the metastasis of human breast and colon cancer cells in SCID mice: 23 out of 26 HPA-positive cancers metastasized, as opposed to only two out of 38 HPA-negative cancers. This experimental model is well suited for investigating the functional role of carbohydrate residues recognized by HPA in breast and colon cancer metastasis. PMID- 9413742 TI - Optimization of PCR/lambda exonuclease-mediated synthesis of sense and antisense DNA probes for in situ hybridization. AB - In situ hybridization experiments are stringently dependent on the quality of the probes, which should be single-stranded when efficient comparison of signals obtained with antisense and control sense probes are needed. In this report, we describe an optimized synthesis of radioactive single-stranded DNA probes, without vector cloning and requiring a unique polymerization step. The sequence region selected as probe is amplified by polymerase chain reaction in the presence of radiolabelled nucleotides. The sense and antisense probes are then yielded by the action of the lambda bacteriophage exonuclease, which can specifically eliminate one out of the two strands of the amplified fragments. In this way, sense and antisense probes with identical length and specific activity can be generated by selecting the primer to be phosphorylated. We have verified the efficiency of our probes for in situ hybridization of the clusterin transcripts within the peripheral olfactory system, after surgical lesion of its synaptic target. PMID- 9413743 TI - Lectin binding sites on CD34+ human haematopoietic stem cells and lymphocytes from peripheral blood: an ultrastructural post-embedding study. AB - This study was performed to obtain a better insight into the glycosylation pattern of human CD34+ haematopoietic stem cells and lymphocytes from peripheral blood using an ultrastructural post-embedding technique. Lectins applied were derived from Canavalia ensiformis (Con A), Triticum vulgare (WGA), Lycopersicon esculentum (LEA), Limulus polyphemus (LPA), Ulex europaeus-I (UEA-I), Bauhinia purpurea (BPA), Glycine max (SBA), Helix pomatia (HPA), Arachis hypogaea (PNA) and Erythrina cristagalli (ECA). Our results showed almost identical staining patterns with both CD34+ cells and mature lymphocytes from peripheral blood. Con A displayed a prominent reactivity with the nuclear envelope and a weak staining of the plasma membrane. As demonstrated by an elaborate lectin double-labelling technique, WGA revealed an opposite staining pattern. Following neuraminidase treatment of sections, BPA, PNA and SBA exhibited a prominent staining of the plasma membrane in CD34+ cells and lymphocytes as well. Membrane reactivity with HPA was restricted to the majority of lymphocytes, presumably T-lymphocytes. Infrequently occurring dense cytoplasmic (lysosomal) bodies were reactive with a variety of lectins, and a weak diffuse nuclear labelling was observable with LPA, UEA-I, WGA and Con A. It is tempting to speculate that carbohydrate moieties on plasma membranes may be involved in the complex mechanisms characterizing cell-to cell interactions (adhesion) and particularly in the so-called phenomenon of homing. PMID- 9413744 TI - Characterization of three new membrane structures on rat NK cells which are involved in activation of the lytic machinery. AB - In this study, three membrane structures on rat NK cells which activate lysis of target cells were characterized. Furthermore, the role of adhesion molecules in this activation process, in particular the CD18-associated integrins, was investigated. Three rat NK-activation structures were identified which have not been previously described. These structures are apparently unique as they differed in molecular weight from known NK-activation structures. Cross-linking of these activation structures with specific mAbs and a Fc gamma R-positive tumor cell line (P815) resulted in enhanced killing of these target cells by NK cells. If the CD18-associated integrins were masked by the anti-CD18 mAb WT.3, the redirected killing of P815 was completely blocked. This indicates that the CD18 associated integrins play a crucial role in activation of NK cells. Furthermore, our results show that rat NK cells possess multiple activation structures. PMID- 9413745 TI - Additive cytotoxicity of different monoclonal antibody-cobra venom factor conjugates for human neuroblastoma cells. AB - Insufficient numbers of antigen molecules and heterogeneity of antigen expression on tumor cells are major factors limiting the immunotherapeutic potential of the few clinically useful monoclonal antibodies capable of mediating complement cytotoxicity and antibody-dependent cellular cytotoxicity. To overcome this limitation, we converted two non-cytotoxic monoclonal anti-neuroblastoma antibodies, designated 3E7 (IgG2b) and 8H9 (IgG1), and the non-cytotoxic F(ab')2 fragment of the cytotoxic monoclonal anti-GD2 antibody 3F8 (IgG3) into cytotoxic antibody conjugates by covalent attachment of cobra venom factor (CVF), a structural and functional homologue of the activated third component of complement. Competitive binding experiments confirmed the different specificities of the three antibodies. In the presence of human complement, all three antibody CVF conjugates mediated selective complement-dependent lysis of human neuroblastoma cells. Consistent with the kinetics of the alternative pathway of complement, approximately seven hours incubation were required to reach maximum cytotoxicity of up to 25% for the 3E7-CVF conjugate, up to 60% for the 8H9-CVF conjugate, and up to 95% for the 3F8 F(ab')2-CVF conjugate. The different extent of maximal cytotoxic activity of the three conjugates was reflected by corresponding differences in the extent of binding of both unconjugated antibodies and the respective conjugates. Any combination of the three antibody CVF conjugates caused an additive effect in complement-mediated lysis. Using a cocktail of all three conjugates, the extent of complement-mediated killing could be increased up to 100%. These data demonstrate that by coupling of CVF the relative large number of non-cytotoxic monoclonal anti-tumor antibodies of interesting specificity can be used to design cocktails of cytotoxic conjugates and, thereby, to overcome the problem of insufficient and heterogeneous antigen expression on tumor cells for immunotherapy. PMID- 9413746 TI - Induction of donor-specific T cell anergy by portal venous injection of allogeneic cells. AB - The mechanisms behind tolerance induction by portal venous (pv) injection of allogeneic cells are investigated. When a hematopoietic stem cell (HSC)-enriched population of BALB/c bone marrow was pv injected into C57BL/6 mice, the response of the T cells in the B6 mice to BALB/c alloantigens in mixed lymphocyte reaction (MLR) decreased until day 4 after the injection. Neither clonal deletion of V beta 11+ T cell nor donor-specific suppressor activity was observed. When recipient T cells were separated into CD4+ and CD8+ cells, only the CD8+ cell population showed donor-specific tolerance. The donor cells were trapped and retained in the host liver. MHC class I antigens were highly expressed on the trapped cells whereas class II antigens or B7 costimulatory molecules were not. The tolerance to BALB/c alloantigens in MLR was obtained also by the pv injection of Meth A, a BALB/c-derived sarcoma cell line. However, tolerance was not induced by the pv injection of B7-transfected Meth A cells. In addition to MLR, tolerance was also observed in DTH responses, and this was also due to the unresponsiveness of CD8+ cells to the donor alloantigens. However, the BALB/c-specific DTH responses were not suppressed after the pv injection of B7-transfected Meth A cells. These results strongly suggest that the tolerance induced by pv injection of allogeneic cells is due to clonal anergy generated by the absence of costimulatory signals in the interaction between donor-specific CD8+ T cells and donor hematopoietic cells trapped in the host liver. PMID- 9413747 TI - Differences in angiogenic potential of classically vs alternatively activated macrophages. AB - Macrophages (M phi) are important for angiogenesis during inflammation, wound repair, and tumor growth. However, well-characterized M phi subsets such as IFN gamma-induced, classically activated (ca) M phi or IL-4/glucocorticoid-induced, alternatively activated (aa) M phi have not been thoroughly examined for a positive or negative association with angiogenesis. While caM phi populate early inflammatory reactions and high-turnover granulomas, aaM phi occur in healing wounds and chronic inflammation. In contrast to caM phi-dominated lesions, aaM phi-rich lesions are highly vascularized. In order to determine their angiogenic potential in vitro, these M phi subsets as well as unstimulated control macrophages (coM phi) were analyzed by RT-PCR for mRNA expression of 10 angiogenic factors after 3 and 6 days of culture. Early during activation, caM phi and coM phi expressed equal levels of 8 of 10 angiogenic factors (PDGF-A, MK, TNF-alpha, TGF-beta 1, PDGF-B, HGF, TGF-alpha, IGF-1), while aaM phi showed expression of only 4 of these factors (TGF-beta 1, PDGF-B, HGF, GF-1). After maturation, TGF-alpha and IGF-1 showed a shift in mRNA expression from caM phi to aaM phi resulting in a considerably enhanced expression of these factors in day-6 aaM phi as compared to day-6 caM phi and coM phi while PDGF-A, MK, and TNF-alpha remained suppressed in day 6 aaM phi. In all M phi subsets including controls, mRNA expression of aFGF and bFGF was minimal or absent while TGFG-beta 1, HGF, and ODGF-B were constitutively expressed. In order to functionally integrate angiogenic factor mRNA expression profiles, mitogenic activity of M phi subsets towards microvascular endothelium was assessed by cocultivation. Coculture experiments revealed that endothelial proliferation induced by aaM phi was 3.0 3.5x higher than induced by caM phi. In conclusion, mature aaM phi are well equipped to play an important role in protracted M phi-associated angiogenic processes. Presumably due to expression of predominantly angio-inhibitory cytokines such as TNF-alpha by caM phi but much less by aaM phi, caM phi exhibit only a low angiogenic potential in vitro and in vivo despite considerable expression of angiogenic factor mRNA. PMID- 9413748 TI - MHC-I antigen expression determines sensitivity of hematopoetic progenitor cells as targets for NK cells. AB - Hybrid resistance is suggested to be mediated by NK lymphocytes as effector cells. NK cytotoxicity is triggered by specific NK cells receptors. One group of receptors recognizing MHC-I antigens is predominantly transmitting an inhibitory signal into the cell. However, NK cells have not been shown to recognize hematopoetic progenitor cells directly. In these studies we demonstrate that hematopoetic progenitor cells are sensitive targets for NK recognition. NK cytotoxicity is shown to depend on MHC-I antigen expression on these target cells. Bone marrow-derived hematopoetic progenitor cells from a beta 2 microglobulin-deficient mouse strain exhibit a significant increase of cytotoxic susceptibility compared with the wild type control. CFU-assays reveal an almost complete loss of proliferation after coincubating MHC-I-deficient bone marrow cells with NK cells from the wildtype mouse strain. The examination of H-2K in allorecognition of hematopoetic progenitor cells reveals an increased cytotoxicity after treatment of resistant syngeneic hematopoetic progenitor cells with H-2Kb F(ab)2-antibodies. Also masking of allogeneic hematopoetic progenitor cells with anti-H-2Kd F (ab)2-antibodies results in enhanced NK killing. Thus, hematopoetic progenitor cells are sensitive targets for NK cells and MHC-I antigen complex is the critical structure in NK recognition of hematopoetic progenitor cells. This complex mediates resistance of NK-specific lysis of hematopoetic progenitor cells. PMID- 9413749 TI - Effects of exercise on circulating vascular adhesion molecules in healthy men. AB - Based on previous studies showing an increase in circulating soluble intercellular adhesion molecule-1 (sICAM-1) after exercise, we hypothesized that exercise may also increase serum levels of the vascular cell adhesion molecule-1 (sVCAM-1) and sE-selection. In a prospective controlled clinical trial, serum levels of sE-selectin, sICAM-1 and sVAM-1 were measured before and after two different exercise protocols in healthy untrained men. Lactate levels increased up to 12.7 mmol/L (95% confidence interval: 10.1-15.9) and 3.6 mmol/L (CI: 2.4 4.7) after ergometry and after an one hour endurance exercise at 60% of the maximal work intensity, respectively (p = 0.028 vs baseline and controls). The maximal increase in lymphocyte counts of 106% (CI: 63-146), which was only of short duration, was higher immediately after ergometry as compared to that observed after endurance exercise (p = 0.028). However, the maximal increase in neutrophil counts of 178% (CI: 120-298) which was seen at 2 hours after endurance exercise was higher than that seen after ergometry (p = 0.028). In contrast, only small changes of circulating adhesion molecules were seen immediately after ergometry: sICAM-1 increased by 11% (CI: 4-25; p = 0.028), and similar tendencies were also observed for sVCAM-1 and sE-selectin. No other consistent and time dependent changes of circulating adhesion molecules were observed and all differences and changes were < or = 11%. In sum our study provides evidence that recreational sporting activities in untrained healthy subjects at normal altitude have little influence on serum levels of the circulating vascular adhesion molecules sE-selectin, sVCAM-1 or sICAM-1. PMID- 9413750 TI - Up-regulation of iNOS mRNA expression and increased production of NO in human monoblast cell line, U937 transfected by HTLV-I tax gene. AB - We investigated the mRNA expression of inducible nitric oxide synthase (iNOS) and the production of nitric oxide (NO) in human T cell lymphotropic virus type I (HTLV-I) p40tax-transfected U937 cells, a human monoblast cell line. Transfection of HTLV-I p40tax U937 cells induced up-regulation of iNOS mRNA expression and subsequent NO production. Furthermore, interferon gamma (IFN-gamma) stimulation of HTLV-I p40tax-transfected U937 cells enhanced iNOS mRNA expression and NO production. The kinetics of iNOS mRNA expression and NO production indicated maximal effect at 24 and 48 hours, respectively, after culture with or without IFN-gamma. These findings suggest that HTLV-I p40tax can act as a transactivator of NO production in cells of Mo/M phi lineage. To what extent this mechanism may be involved in the pathogenesis of HTLV-I-associated diseases warrants further investigation. PMID- 9413751 TI - Interleukin-12 enhances CD26 expression and dipeptidyl peptidase IV function on human activated lymphocytes. AB - Research of a cellular pathway activated by IL-12 which may result in new therapeutical approaches for IL-12, led us to find an intriguing relationship between IL-12 and CD26/DPPIV ectopeptidase on activated T cells. Both the percentage and median fluorescence intensity (MFI) of CD26+ cells in the PHA stimulated PBMC or lymphoblasts increased when IL-12 (optimum dose, 2 ng/ml) was present. Maximum CD26 expression was observed on day-2 cultures of lymphoblasts, the presence of IL-12 receptor probably being necessary for this upregulation. In addition, CD26 upregulation correlated with enhanced DPPIV function. Enzyme affinity and secretion of the soluble form of DPPIV were not affected by IL-12. Kinetic behaviours of Ag expression and enzymatic activity support a different CD26 regulation pathway by IL-12. These data suggest that the correlation found in vivo between the CD26 expression and Th1-like immune responses is due to this IL-12-dependent upregulation. PMID- 9413752 TI - Monocyte-derived dendritic cells represent a transient stage of differentiation in the myeloid lineage. AB - Cultivation of human peripheral blood monocytes with granulocyte/macrophage colony stimulating factor (GM-CSF) and IL-4 facilitates generation of strongly antigen-presenting dendritic cells (DC). These monocyte-derived DC (mdDC) were used here to further delineate differentiation pathways in the myeloid lineage. Incubation of mdDC with TNF or soluble CD40L led to enhanced MHC and accessory surface antigen expression with significantly elevated T cell stimulatory activity, indicative of DC maturation. In contrast, after cytokine withdrawal or incubation with M-CSF, mdDC differentiated to macrophages. Cells became adherent, monocyte/macrophage surface markers were upregulated, and MHC and accessory surface proteins were downregulated. Furthermore, the multilaminar MHC class II compartments (MIIC) were lost and the T cell stimulating capacity largely diminished. Thus, mdDC show a high developmental plasticity by retaining their ability to become macrophages or to continue their differentiation towards mature DC. PMID- 9413753 TI - Helper activity of chicken V beta 1+ and V beta 2+ alpha beta T cells for in vitro IgA antibody synthesis. AB - Chickens have only two T cell receptor variable beta gene families: V beta 1 and V beta 2 (1). In our previous work we found that IgA production was almost completely suppressed in chickens depleted of V beta 1+ alpha beta T cells by treatment with a TCR V beta 1-specific monoclonal antibody (2), while IgM and IgG production was not affected. Our present results indicate that, in vitro, both V beta 1+ and V beta 2+ chicken cecal tonsil T cells provide help for the differentiation of cecal tonsil IgA B cells, suggesting that the failure of V beta 1+ T cell-depleted chickens to produce IgA is not caused by the inability of V beta 2+ T cells to provide help for IgA production by B cells, but rather by the scarcity of these T cells in mucosal tissues (3), where most IgA responses are induced (4). PMID- 9413754 TI - Multiple primary tumours of the urogenital system: our cases and a review of the literature. AB - We had 6 genitourinary tumour patients with other primary tumours in a 147 genitourinary tumour patient group. Two of these tumours developed synchronously while the others were metachronous, and two of them developed in the genitourinary system. It is clear that cancer patients have a higher risk of a second cancer than the general population. Second cancers may be detected in long term follow-up of cancer patients, or may be caused by the aetiologic factor(s) with pleiotropic effects, or may develop because of the therapies for the first cancer. PMID- 9413755 TI - The necessity of prophylactic antibiotics during extracorporeal shock wave lithotripsy. AB - In this study we treated 340 patients with renal and ureteric stones. They all underwent ESWL with the HM-4 lithotriptor. The patients were divided into two groups, the first one including 250 patients and the second 90. The first group consisted of patients with sterile urine prior to ESWL. These patients did not receive any antibiotic prophylaxis, while 5.2% of them developed infectious problems which were followed by significant bacteriuria in only 2% of the cases. The 90 patients of the second group had urinary tract infection on the preoperative cultures and received antibiotic treatment. Of these patients 27.8% developed infectious problems which were followed by significant bacteriuria in 21.1% of the cases. Evaluating the above results, we estimate that the administration of prophylactic antibiotics in the case of patients with sterile urine before ESWL is not required while it may prove to be useful in the case of patients with urinary tract infection prior to ESWL. PMID- 9413757 TI - Biliary peritonitis complicating percutaneous nephrostomy. AB - Biliary peritonitis is a rare but serious complication of percutaneous renal access and surgery. We present such a case and review the literature. PMID- 9413756 TI - Markers of bone turnover in patients with nephrolithiasis. AB - A total of 19 patients with active nephrolithiasis, 14 patients with non-active nephrolithiasis and 17 healthy subjects were examined under standardized intake of calcium, phosphorus, purine and protein. In patients with both active and non active renal stone disease the following abnormalities were found: elevated plasma levels of PTH and osteocalcin, increased activity of the bone isozyme of alkaline phosphatase, low plasma levels of phosphate and increased urinary excretion of calcium and oxalic acid. These abnormalities were more marked in patients with active than non-active nephrolithiasis. No correlation was found between plasma PTH levels and parameters of bone turnover as well as calciuria and oxaluria. Results presented in this paper suggest that (a) Smith's criteria of active renal stone disease are of minor pathogenetic and therapeutic value and (b) patients with active nephrolithiasis differ from non-active renal stone formers by more elevated oxaluria and markers of bone turnover and more marked abnormalities in calcium-phosphate metabolism related parameters. PMID- 9413759 TI - A case of ureteral triplication (type 1) associated with vesicoureteral reflux in a solitary kidney. AB - Ureteral triplication is a rare congenital anomaly of the upper urinary tract. It is reported to be associated with an increased incidence of congenital anomalies as well as a predisposition to infection and calculus formation. We report a case of type 1 variant of ureteral triplication associated with vesicoureteral reflux into lower and mid pole ureters in a solitary kidney. To our knowledge ureteral triplication in a solitary kidney has not been described previously. PMID- 9413758 TI - Adult Wilms tumour. A report of two cases and their treatment and prognosis. AB - Adult Wilms tumour, unlike that seen in childhood, is a rare disease. We are reporting two cases of which one is alive with no evidence of disease at the 52nd month of follow-up and the other had no evidence of disease at the 10th month after which she was lost to follow-up. The literature has been reviewed and the prognosis and treatment alternatives have been surveyed. PMID- 9413760 TI - Ureteric obstruction due to B-cell lymphoma. PMID- 9413761 TI - Fibroepithelial polyp of the ureter associated with an adjacent ureteral calculus. AB - Ureteral fibroepithelial polyps are rare benign tumours. Sometimes an ureteric calculus may be seen, proximal to the polyp. We describe a case of fibroepithelial polyp (FEP) associated with a calculus in the distal part of the right ureter. To our knowledge this is the first ureteral FEP reported in the literature associated with a ureteral calculus distal to it. Aetiology, clinical features, diagnosis and therapy of fibroepithelial polyps are discussed in the light of the literature. PMID- 9413762 TI - Thermoreceptor mediated bladder sensation in patients with diabetic cystopathy. AB - Urinary bladder sensation to ice water instillation was evaluated in 32 patients with diabetes mellitus. Sixteen patients were urodynamically normal (group 1) and the remaining 16 were diagnosed as having diabetic cystopathy (group 2). Eleven out of 16 patients in group 2 could be considered as having impairment of sensation to urinate. Two out of 16 (12.5%) subjects in group 1 could not perceive ice water infusion, whereas 25% of patients in group 2 did not feel the ice water sensation. In group 2, patients with impaired urinary sensation showed relatively high incidence of negative ice water test (36.4%), although there were no statistical differences between the other groups. There was no apparent relationship between prevalence of peripheral neuropathy and that of negative ice water test. Impairment of ice water perception was less frequent than that of mechanoreceptor sensation in patients with diabetic cystopathy. PMID- 9413763 TI - Intravesical bacillus Calmette-Guerin (Tokyo 172 strain) therapy for carcinoma in situ of the bladder. AB - To evaluate the clinical response to intravesical instillation therapy with bacillus Calmette-Guerin (BCG) Tokyo 172 strain for carcinoma in situ (CIS) of the bladder and subsequent patient prognosis, we reviewed data for 30 patients treated between 1985 and 1994. Median follow-up was 56 months. The CIS cases comprised two groups: primary (19 patients) and subsequent to development of a gross neoplasm (11 patients). Either 40 mg (n = 20) or 80 mg (n = 10) doses of BCG were instilled weekly for 8 weeks. This intravesical therapy resulted in apparent eradication of tumour cells in 25 of the 30 patients for a complete response (CR) rate of 83%, with no difference found between primary and secondary groups. Tumours later recurred in 6 of the 25 patients (24%) and disease progression was found in only 3 (12%). In contrast, progression occurred in 3 of 5 patients (60%) for which a complete response was not achieved with intravesical BCG therapy. The difference between these two groups was significant (p = 0.04). Total cystectomy was performed in 2 of 25 CR patients (8%) first and in 4 of the 5 unresponsive (80%), the statistical difference being highly significant (p = 0.003). The 5-year survival rate was 96% for the study subjects as a whole. In conclusion, CIS unresponsive to BCG therapy should be treated with immediate total cystectomy while cases demonstrating a complete response should be followed up for a long period. PMID- 9413764 TI - Zinc in the human prostate gland: normal, hyperplastic and cancerous. AB - Zinc concentration in a prostate gland is much higher than in other human tissues. Data for zinc changes in different prostate diseases are limited and greatly contradictory. To analyze transrectal puncture tissue biopsy and resected materials, zinc content was estimated in benign prostatic hyperplasia (BPH) and cancer. There were 109 patients studied (50 BPH and 59 cancer). The control group consisted of 37 intact glands of men who died an unexpected death (accident, murder, acute cardiac insufficiency, etc.). All materials studied were divided into two parts. One of them was morphologically examined, while the zinc content of another one was estimated. The radionuclide induced energy dispersive X-ray fluorescent analysis was used for zinc determination. Zinc content (M +/- SE) of normal prostate, BPH and cancer was 1018 +/- 124, 1142 +/- 77, and 146 +/- 10 micrograms/g dry tissue, respectively. It was shown that zinc assessment in the material of transrectal puncture biopsy of prostate indurated site can be used as an additional test for differential diagnosis of BPH and cancer. Accuracy, sensitivity and specificity of the test are 98 +/- 2%. PMID- 9413765 TI - Effects of distilled water and mixture of sorbitol-mannitol irrigation fluids on fluid-electrolyte balance in patients undergoing transurethral prostatectomy. AB - This study aimed to compare two various irrigation fluids used in transurethral prostatectomy (TURP) with respect to changes in fluid-electrolyte balance and to evaluate the blood loss during TURP. TURP was performed in 50 patients using distilled water and in 42 patients using a mixture of sorbitol and mannitol solution (2.7% sorbitol and 0.54% mannitol) as irrigation fluid. Fluid electrolyte changes and blood loss were evaluated. None of the patients had transurethral resection syndrome. Decline in serum sodium level was more significant in patients who were operated on with the sorbitol plus mannitol irrigation fluid in contrast to its clinical insignificance (p < 0.05). Decrease in serum sodium concentration in patients with more than 15 g of tissue resected was greater than in patients with less than 15 g of tissue resected (p < 0.05). Blood loss was greater in the distilled water group (p < 0.05). Blood loss was 145.5 +/- 3.4 ml in patients given a blood transfusion. In conclusion, a fluid containing mainly sorbitol must be the irrigation fluid preferred over distilled water because of its non-haemolytic nature, but although not important clinically, it can cause hyponatraemia more than distilled water. Furthermore, it is useful to determine blood loss during TURP before deciding to give a blood transfusion. PMID- 9413766 TI - Rete testis adenocarcinoma recurring in the inguinal lymph nodes. A case report. AB - Adenocarcinoma of the rete testis is a rare neoplasm with 41 reported cases in the literature till 1994. In most of the reported cases, the neoplasm presents as a scrotal mass with diffuse enlargement. The aetiology is unknown and the clinical course of the tumour is not very well defined. In six of the reported cases metastatic spread of the tumour to inguinal lymph nodes was demonstrated in the follow-up. We report herein a distinctive case of rete testis adenocarcinoma presenting as an isolated inguinal recurrence one year after radical orchiectomy. PMID- 9413767 TI - Testis sparing surgery for epidermoid cyst of the testis: a case report. AB - A rare tumour of a prepubertal child, an epidermoid cyst, was excised with testicular preservation. Childhood testicular tumours are usually benign. Although epidermoid cysts of the testis may have teratomatous component, testicular teratomas are generally benign in the prepubertal child. For these reasons testis sparing surgery seems applicable in childhood epidermoid cysts. PMID- 9413768 TI - Testicular needle biopsy: is it a safe and adequate method? AB - We aimed to investigate if testicular needle biopsy is adequate and safe for the examination of testis. Needle biopsies were performed on 21 testes of 5 patients with advanced prostate cancer and 11 patients with cryptorchidism before orchiectomy. Biopsies were done with the prostatic tru-cut needle. After needle biopsy, the tract and puncture sites were explored and an incisional biopsy was performed on each testis. Both needle specimens and open biopsy specimens were fixed in Bouin's solution and sent for histologic examination. There were small haematomas in two testes and moderate haemorrhage between tunica vaginalis layers in another. The tissues obtained by needle biopsy were sufficient except for two specimens and diagnostic accuracy was perfect. Nevertheless, measurement of the seminiferous tubules of the lamina propria could not be achieved in many cases. PMID- 9413769 TI - Plasma lipoproteins in patients with chronic renal failure (CRF). AB - The clinical picture in chronic renal failure (CRF) shows great variability depending on age, sex, aetiology of disease, grades of renal injury and type of treatment. Significant increases of triglycerides (TG), low-density lipoprotein cholesterol (LDL-chol) and and apo B concentrations, significant decreases of high-density lipoprotein cholesterol (HDL-chol) levels and apo A and apo AI concentrations, and no significant changes in total cholesterol (TC) have been shown in CRF patients. Significant increases of TC/HDL-chol, LDL-chol/HDL-chol, apo B/apo AI and apo B/LDL-chol ratios were also demonstrated. That indicates a high risk of atherosclerosis even when total cholesterol levels are in the normal range. There were highly significant and positive correlations between TC/HDL chol and LDL-chol/HDL-chol ratios, apo B and LDL-chol concentrations as well as between the apo B/apo AI and LDL-chol/HDL-chol ratios. PMID- 9413770 TI - The effect of metabolic acidosis on serum apolipoprotein A I and apolipoprotein B levels in children with chronic renal failure. AB - In this report serum apolipoprotein A I (Apo A I) and apolipoprotein B (Apo B) levels were determined in children with chronic renal disease (CRD) during metabolic acidosis, after the correction of metabolic acidosis and in healthy children to look for the effect of metabolic acidosis on Apo A I and B levels. It was found that Apo A I levels were significantly decreased during metabolic acidosis (p < 0.05) but Apo A I/Apo B ratios were not affected before and after the correction of acidosis in the CRD group (p > 0.05) although it was significantly different from those in the controls (p < 0.01). PMID- 9413771 TI - The role of endothelin in radiocontrast nephropathy. AB - In the present study we investigated the role of endothelin and AT II in radiocontrast nephropathy induced in rats with reduced renal mass (70-75%). Thirty-five male Wistar albino rats weighing between 280 and 400 g were anaesthetized with ketamine (130 mg/kg b.w.) and right total, left 50% nephrectomy were performed. After this operation, the rats were kept under observation for six to eight weeks and then they were randomly separated into three groups. Group I rats were infused with 8.9 ml/kg (or 2.9 g of iodine/kg body weight) Na diatrizoate (Urovision, 1,500 mosm/kg). Group II rats were infused with 0.9% NaCl in an equal volume with the radiocontrast material. Group III rats were given 4.5% NaCl that had the same volume and osmolality as the radiocontrast material. Two hours after the drug infusions, blood and accumulated urine samples were collected from all the rats and tested for endothelin, AT II, BUN, creatinine, uric acid, electrolytes, calcium and phosphorus. We found that the plasma endothelin levels in Group I (77.64 +/- 29.62 pg/ml) were significantly higher than in Group II (20.52 +/- 5.83 pg/ml) and Group III (15.04 +/- 5.15 pg/ml) (t = 8.34 and t = 9.14, respectively, p < 0.001). Therefore elevation in circulating endothelin might have been an additional factor leading to the radiocontrast-induced nephrotoxicity. PMID- 9413772 TI - Haematoma block or Bier's block for Colles' fracture reduction in the accident and emergency department--which is best? AB - OBJECTIVE: To offer clear guidance on the anaesthetic management of Colles' fractures in the accident and emergency (A&E) department in the light of the conflict between existing reports and current trends, and to address the issue of alkalinisation of haematoma blocks. METHODS: This was a two centre, prospective, randomised clinical trial with consecutive recruitment of adult patients with Colles' fractures requiring manipulation to receive either Bier's block or haematoma block. There was subsequent blinded randomisation to alkalinised or non alkalinised haematoma block. RESULTS: 72 patients were recruited into the Bier's block group, and 70 into the haematoma block group. Bier's block was less painful to give than the haematoma block (median pain score 2.8 v 5.3; P << 0.001), and fracture manipulation was also less painful in the Bier's block group (median pain score 1.5 v 3.0; P < 0.01). There was no significant difference in overall A&E transit time between the two groups. There was better initial radiological outcome in terms of dorsal angulation in the Bier's block group (-3.6 degrees v 2.1 degrees; P = 0.003). More remanipulations were required in the haematoma block group (17/70 v 4/72; P = 0.003). There was a trend towards decreased pain on administration of the alkalinised haematoma block when compared with non alkalinised haematoma block, but this did not reach significance. There was no difference in pain score on fracture manipulation. There were no complications in either group. CONCLUSIONS: Bier's block is superior to haematoma block in terms of efficacy, radiological result, and remanipulation rate; transit times are equal, both procedures are practical in the A&E environment, and there were no complications. Bier's block is the anaesthetic management of choice for Colles' fractures requiring manipulation within the A&E department. PMID- 9413773 TI - Use and effect of paediatric advanced life support skills for paediatric arrest in the A&E department. AB - OBJECTIVES: To define the use of paediatric advanced life support by the Leicestershire Ambulance and Paramedic Service (LAPS) and the A&E department of a large university teaching hospital; and to identify the outcome and determine the factors that are consistent with a successful outcome. SUBJECTS AND METHODS: The prehospital, accident and emergency (A&E), and inpatient notes of all patients aged 0-16 years who had been admitted to the resuscitation room at the Leicester Royal Infirmary in cardiac arrest between 1 January 1992 and 31 December 1995 were reviewed. Cardiac arrest was defined according to the Utstein template for reporting of prehospital data. RESULTS: During the four year period, 51 cases of paediatric cardiac arrest were identified, with a median age of 3.2 years (range two days to 15 years). In eight patients, resuscitation was not attempted. Of the remaining 43, 15 (37%) were discharged from A&E to the intensive care unit. Five (11.5%) ultimately survived to discharge from hospital. Subsequent neurological development was recorded as normal in four of the five. Of the patients who had a prehospital cardiac arrest and were initially resuscitated by the LAPS there was only one survivor. He was discharged from hospital with severe neurological injury and died three months later. CONCLUSIONS: The outcome for established prehospital paediatric cardiac arrest, in a well defined emergency medical services system, is very poor at present. It does not seem to be affected by the institution of paediatric life support teaching programmes for hospital staff alone. The timing in instituting advanced life support measures remains the most critical factor affecting outcome in these patients. PMID- 9413774 TI - Implementation of the Ottawa Ankle Rules by nurses working in an accident and emergency department. AB - OBJECTIVE: To assess whether accident and emergency (A&E) nurses using the Ottawa Ankle Rules could detect all ankle fractures. DESIGN: Prospective observational study. SETTING: A&E department of a university teaching hospital. SUBJECTS: All patients who presented with ankle injuries who were initially assessed by a nurse taught the Ottawa Ankle Rules. OUTCOME MEASURES: (1) The numbers of patients referred by the nurse for ankle radiography; (2) of these, the number with ankle fractures; (3) of those not sent for radiography initially by the nurse, the number who subsequently had x rays (ordered by the doctor) and had a fracture; (4) of those having no x rays, the number who reattended later. RESULTS: 324 patients were eligible; 238 had x rays at the request of the nurse (73%); 48 of these (20%) were diagnosed as having a fracture. Of those 86 patients not sent for radiography by the nurse, 19 subsequently had x ray examinations at the request of a doctor and no fracture was detected. Of the 67 not sent for radiography, none returned within the subsequent eight weeks. CONCLUSIONS: Nurses can apply the Ottawa Ankle Rules safely without missing acute fractures; that is, of those who were not sent for radiography by nurses, none subsequently reattended the A&E department or the trauma service of the Bristol Royal Infirmary during the following two months. PMID- 9413775 TI - Should relatives be allowed in the resuscitation room? AB - OBJECTIVE: To assess doctors' and nurses' views on the presence of relatives in the resuscitation room during cardiac arrest or major trauma. DESIGN: Questionnaires were sent to accident and emergency (A&E) nurses and doctors of all disciplines in a London teaching hospital. Recipients were asked if they would favour the presence of selected relatives in the resuscitation room and to give comments. RESULTS: 103 questionnaires were distributed and 81 returned, a response rate of 78.6%; 33% were senior house officers, 29% consultants, 16% senior registrars/registrars, 12% A&E nurses, and 10% house officers. Of the respondents, 63% were not in favour of relatives being present, and 37% were in favour. The likelihood of being in favour of allowing relatives to be present was high among A&E nurses; among doctors it increased with rising seniority. Most respondents felt that more resuscitation training would be necessary, in addition to counselling for staff and relatives. CONCLUSIONS: Staff with the least experience in dealing with resuscitations and distressed relatives were likely to be opposed to relatives being present in the resuscitation room. As there is evidence that the bereavement process is eased if a partner/relative witnesses the resuscitation, relatives should be offered the opportunity to witness resuscitation if staff training is geared towards the presence of relatives. ALS/ATLS training for all hospital doctors and nurses should include the management of distressed relatives observing a resuscitation. PMID- 9413776 TI - How do individuals with diabetes use the accident and emergency department? AB - OBJECTIVE: To determine whether the frequency and pattern of use of the accident and emergency (A&E) department by individuals with diabetes is different from that of the general population. METHODS: A historical cohort of 696 individuals with diabetes from six randomly selected general practices and a non-diabetic comparison cohort matched on age, sex, and general practice were identified. The use of an urban A&E department by the two cohorts was compared for number of visits between 1984 and 1996 for injuries, diabetes related and non-diabetes related illness, proportion referred by a general practitioner, proportion arriving by ambulance, and proportion admitted. RESULTS: More visits were made by the diabetic cohort (1002 v 706, P = 0.0001); 121 visits were directly related to diabetes, including 52 for hypoglycaemia. The diabetic cohort also had more visits for medical illness unrelated to diabetes (357 v 231, P = 0.0001). The number of visits for injuries was similar (524 v 475, P = 0.3). Individuals with diabetes who attended A&E were not significantly more likely to be referred by a general practitioner (14% v 16%) or admitted (20% v 17%). CONCLUSIONS: Individuals with diabetes made more frequent visits than the general population to the A&E department. Since there was no excess of visits for injuries and the proportion requiring admission was similar, the hypothesis that they have a different threshold for attending is not supported. PMID- 9413777 TI - Characteristics of female victims of assault attending a Scottish accident and emergency department. AB - OBJECTIVE: To compare the characteristics of female victims of assault with those of male victims and to see if there is a difference between female victims of domestic assault and females assaulted by strangers or acquaintances. DESIGN: A two month prospective study (June and July 1995) of all assault victims attending a Scottish accident and emergency (A&E) department. SETTING: A large district general A&E department (the Royal Alexandra Hospital in Paisley) seeing 60,000 new patients per year with a catchment population of 200,000. RESULTS: 46 female victims of violence attended the A&E department (20% of the total of 235). In comparison with men, women were more likely to be assaulted in their homes (48% v 10%; P < 0.001), but were less likely to be assaulted with sharp weapons (7% v 28%; P = 0.003) and to require admission to hospital (P = 0.005). Nineteen women (41%) were victims of domestic assault. The victims of domestic assault were more likely to have been drinking (11% v 31%; P = 0.007) and to have a history of previous assault (63% v 22%; P = 0.002). This group also had a higher mean deprivation score and rate of unemployment, although the differences were not statistically significant. CONCLUSIONS: A&E staff should be aware of risk factors associated with domestic assault to aid recognition of victims. Using the current British Association for Accident and Emergency Medicine guidelines on domestic violence and closer liaison with police, social services, and general practitioners will help prevent further attacks. PMID- 9413778 TI - Treatment of accidental digital injection of adrenaline from an auto-injector device. AB - Following an increase in the number of patients attending the accident and emergency department because of accidental injection of adrenaline from autoinjector devices prescribed for patients with severe allergic reactions, a review of published reports was undertaken to identify the best form of treatment. Local injection of phentolamine is effective for up to 13 hours after the inadvertent digital instillation of adrenaline. PMID- 9413780 TI - Post-traumatic stress disorder. PMID- 9413781 TI - Accident and emergency medicine--making waves on the Internet. AB - The internet is a communications and information tool which has recently entered the world of accident and emergency (A&E) medicine. It is a worldwide instrument facilitating the dissemination of ideas and clinical information in the specialty. It is being embraced by all disciplines involved in A&E medicine. Part I introduces the internet to those in A&E medicine unfamiliar with this technology. It describes the varied resources of the internet in A&E medicine and speculates on its future role. Part II supplies the reader with the necessary information to get on-line and explains some of the more technical aspects of the internet. PMID- 9413779 TI - The value of current developments in radiology to the accident and emergency department--a pictorial review. PMID- 9413782 TI - Fish tank granuloma--a frequently misdiagnosed infection of the upper limb. AB - Five patients attended the accident and emergency (A&E) department with fish tank granuloma caused by an infection with Mycobacterium marinum. All patients had forearm symptoms which were initially misdiagnosed. They were later recognised by the presence of superficial cutaneous lesions in a sporotrichotic distribution. Definitive diagnosis was confirmed by the histological appearances of a biopsy and or culture of the organism. All patients responded to oral minocycline and had uncomplicated recoveries once the diagnosis was established. A&E doctors need to be aware of the possible diagnosis of fish tank granuloma especially when treating forearm infections which have been resistant to antibiotics. PMID- 9413783 TI - Regional intravenous calcium--an effective method of treating hydrofluoric acid burns to limb peripheries. AB - An effective method of providing pain relief in hydrofluoric acid burns is reported, using a Bier's block type technique and regional intravenous calcium gluconate. This method allows satisfactory analgesia and prevents further tissue destruction, without the risk and added discomfort of increased tissue tensions associated with local infiltration of calcium. PMID- 9413784 TI - Use of A&E wards. PMID- 9413785 TI - Gamma hydroxybutyrate poisoning. PMID- 9413786 TI - Return of the bot fly. PMID- 9413788 TI - Annotation: structural equation models in developmental research. PMID- 9413787 TI - Dermatobia--tropical myiasis. PMID- 9413790 TI - Early reading difficulties and later conduct problems. AB - The relationships between early reading difficulties and later conduct problems were examined in a birth cohort of New Zealand children studied from the point of school entry to the age of 16. Children with early reading difficulties had increased rates of conduct problems up to the age of 16 years. These associations depended on context, being more evident for boys and tending to reduce with increasing age. However, the associations between early reading difficulties and later conduct problems were explained by the fact that children with early reading difficulties tended to be characterised by a number of disadvantageous features (and notably early-onset conduct problems) that were present before the onset of reading difficulties. When the associations between reading difficulties and conduct problems were adjusted for confounding factors there were no statistically significant associations between reading difficulties and conduct problems. These results were found to hold for various age and gender subgroups of the sample, for measures of reading difficulties defined in different ways, and for a wide range of outcome variables. It is concluded that, when due allowance is made for potentially confounding factors (and notably early conduct problems) and for factors correlated with these problems, it is unlikely that reading difficulties in early childhood are related to later conduct problems. PMID- 9413789 TI - Practitioner review: physical investigations in mental retardation. AB - Mental retardation occurs in more than 1% of the child population. A cause can be found in almost 80% of individuals with severe mental retardation, but in fewer than 40% of those with mild mental retardation. A work-up is indicated in all cases of mental retardation. A medical doctor with specific training in the field is needed to make "decision-tree diagnosis" and to suggest the most appropriate physical investigations in each case. This paper provides practical guidelines for diagnosis and work-up both in severe and mild mental retardation. PMID- 9413791 TI - Vulnerable adolescent girls: opposite-sex relationships. AB - This interview-based study compares the opposite-sex relationships of 50 girls, aged 15-16, identified as being at risk for difficulties in early adult partnerships, with 50 girls of the same age from an inner-city school. The high risk girls had begun solo-dating earlier than the school girls, were more likely to have had a sexual relationship, to have had more sexual partners, to have been pregnant, and to have had a child. A third of the girls in both groups were solo dating at the time of the interview. In contrast to the school girls, the high risk girls attached a prominence and permanence to their current dating relationships, which already bore the hallmarks of later unsupportive partnerships. PMID- 9413792 TI - An investigation of adolescents' substance use behaviors, depressed affect, and suicidal behaviors. AB - Data from a four-wave panel design of 975 adolescents were used to study inter relationships among suicidal behaviors, depressive symptoms, and substance use behaviors. Persistently high levels of problem drinking and depressive symptoms were associated with higher levels of suicidal thoughts and attempts. Higher levels of depressive symptoms and greater cigarette and illicit drug use distinguished suicidal ideators from attempters. Adolescents attempting suicide reported lower levels of family social support, a greater use of substances to cope with stressors, and a higher density of substance-using peers. Implications of the findings for preventive interventions with high-risk teens are discussed. PMID- 9413793 TI - Attention deficit hyperactivity disorders and other psychiatric outcomes in very low birthweight children at 12 years. AB - One hundred and thirty-seven very low birthweight (VLBW) children were compared at 12 years with a sample of matched peers on a number of psychiatric symptoms including Attention Deficit/Hyperactivity Disorder, depression, anxiety, and antisocial behaviour using the Child and Adolescent Psychiatric Assessment parent interview and various parent and child questionnaires. The main psychiatric risk was Attention Deficit Hyperactivity (ADH) disorders, with 31/136 (23%) VLBW children meeting clinical criteria, compared to 9/148 (6%) of peers. VLBW children were also more likely to have generalised anxiety and more symptoms of depression. More than one quarter of VLBW children (38/136; 28%) showed a psychiatric disorder of some type compared to 9% (14/148) of peers. VLBW children are at increased risk of psychiatric symptoms especially ADHD. This outcome is discussed in relation to neurological, demographic, and cumulative impairment factors. PMID- 9413794 TI - Genetics and developmental psychopathology: 1. Phenotypic assessment in the Virginia Twin Study of Adolescent Behavioral Development. AB - We introduce an overlapping cohort sequential longitudinal study of behavioral development and psychopathology in a representative sample of 1412 pairs of twins aged 8 through 16 years. Multiple phenotypic assessments involve a full psychiatric interview with each child and each parent, and supplementary parental, teacher, and child interview material and questionnaires. For the first wave of assessments, the numbers of reported DSM-III-R symptoms of Major Depressive Disorder (MDD), Separation Anxiety Disorder (SAD), Overanxious Disorder (OAD), Oppositional Defiant Disorder (ODD), Conduct Disorder (CD), and Attention Deficit Hyperactivity Disorder (ADHD), assessed through interviews, confirm patterns of age and sex trends found in other epidemiological samples, but underscore their dependence on whether the child or the parent is the informant. Correlations across domains for symptoms reported by the same informant are often as large as correlations across informants for the same domain of symptoms. Factor analyses of these symptom counts, taking account of informant view and unreliability of assessment, show the high degree of correlation between SAD and OAD, between MDD and OAD, and between CD and ODD. ADHD symptoms are relatively independent of the other domains, but show moderate correlations with CD, ODD, and MDD. Factorially derived dimensional questionnaire scales, based on child, parental, and teacher reports, show patterns of relationship to symptom counts consistent with both convergent and discriminant validity as indices of liability to clinical symptoms. Across informants, questionnaire scales provide as good a prediction of symptoms as do clinical interviews. Multitrait-multimethod confirmatory factor analysis reveals the patterns of relationship between symptoms of psychiatric disorder in children taking due account of informant and unique sources of variance. Gender differences are consistent within the correlated clusters of ODD/CD and MDD/SAD/OAD, although there are disorder-specific age trends. There are large informant-specific influences on the reporting of symptoms in clinical interviews. Dimensional questionnaire scales provide a useful source of additional information. In subsequent analyses of genetic and environmental etiology of childhood psychopathology we must expect that results may differ by informant and method of assessment. Multivariate and developmental analyses that explore the sources of these differences will shed new light on the relationship between genetic and environmentally influenced vulnerability and the manifestation of psychopathology in specific circumstances. PMID- 9413795 TI - Genetics and developmental psychopathology: 2. The main effects of genes and environment on behavioral problems in the Virginia Twin Study of Adolescent Behavioral Development. AB - Little is known about the contribution of genetic and environmental factors to risk for juvenile psychopathology. The Virginia Twin Study of Adolescent Behavioral Development allows these contributions to be estimated. A population based, unselected sample of 1412 Caucasian twin pairs aged 8-16 years was ascertained through Virginia schools. Assessment of the children involved semi structured face-to-face interviews with both twins and both parents using the Child and Adolescent Psychiatric Assessment (CAPA). Self-report questionnaires were also completed by parents, children, and teachers. Measures assessed DSM-III R symptoms of Attention Deficit Hyperactivity Disorder (ADHD). Conduct Disorder, Oppositional Defiant Disorder, Overanxious Disorder, Separation Anxiety, and Depressive Disorder. Factorially derived questionnaire scales were also extracted. Scores were normalized and standardized by age and sex. Maximum likelihood methods were used to estimate contributions of additive and nonadditive genetic effects, the shared and unique environment, and sibling imitation or contrast effects. Estimates were tested for heterogeneity over sexes. Generally, monozygotic (MZ) twins correlated more highly than dizygotic (DZ) twins, parental ratings more than child ratings, and questionnaire scales more highly than interviews. DZ correlations were very low for measures of ADHD and DZ variances were greater than MZ variances for these variables. Correlations sometimes differed between sexes but those for boy-girl pairs were usually similar to those for like-sex pairs. Most of the measures showed small to moderate additive genetic effects and moderate to large effects of the unique individual environment. Measures of ADHD and related constructs showed marked sibling contrast effects. Some measures of oppositional behavior and conduct disorder showed shared environmental effects. There were marked sex differences in the genetic contribution to separation anxiety, otherwise similar genetic effects appear to be expressed in boys and girls. Effects of rater biases on the genetic analysis are considered. The study supports a widespread influence of genetic factors on risk to adolescent psychopathology and suggests that the contribution of different types of social influence may vary consistently across domains of measurement. PMID- 9413797 TI - Normative and pathological obsessive-compulsive behavior and ideation in childhood: a question of timing. AB - This study was designed to test whether obsessive-compulsive behavior declines over development. A cross-sectional design was used on a large community sample of children. Children in grades four, six, and eight (N = 1083), 8 to 14 years of age, were administered the Maudsley Obsessive Compulsive Inventory (MOCI) and the Spielberger State Trait Anxiety Scales. Whereas the mean MOCI score was significantly lower in the eighth grade than in the sixth and fourth, there was an elevation of children with very high MOCI scores in the eighth grade. Obsessive ideas and compulsive behaviors that were common for fourth-grade children were present in only a minority of children in the eighth grade, and were associated with high levels of anxiety. No gender differences were observed for overall obsessive-compulsive behavior, but checking behavior was higher in boys, and cleaning behavior in girls. State anxiety was higher in girls than in boys, and was also higher in older than in younger children. PMID- 9413796 TI - Maternal depressive symptoms and ratings of emotional disorder symptoms in children and adolescents. AB - This study uses information collected on two occasions from a probability sample of families with 8- to 12-year-old children (N = 718) participating in a general population study in 1983 and follow-up in 1987. It focuses on the association between maternal depressive symptoms and emotional disorder in children and adolescents, taking into consideration the influences of informant rating errors, contextual variables (economic disadvantage and family dysfunction), and child gender. Covariance structure analysis revealed a strong association between maternal depressive symptoms in girls (beta = .59 in 1983 and beta = .39 in 1987) but not in boys. This association is independent of the impact of contextual variables and the treatment of teacher rating errors. Among adolescent boys, maternal depressive symptoms are correlated with mother rating errors, suggesting the possible presence of maternal bias. PMID- 9413798 TI - Can young people with autism refer to knowledge states? Evidence from their understanding of "know" and "guess". AB - A number of studies have reported that most individuals with autism have difficulty in attributing mental states. The primary aim of the present study was to compare the ability of children with autism to refer to knowledge states with that of mainstream school children and children with Down's syndrome. The second aim was to investigate the role of verbal mental age in referring to knowledge states. The third aim was to compare the ease with which the children referred to their own mental state and to that of others. The results suggest that some individuals with autism are able to attribute knowledge to themselves and others but that they need to have higher verbal skills than is necessary in normal individuals. The level of language skill predicted the performance of the individuals with autism, but not that of the children in the other groups. There was no good evidence that referring to one's own mental states was easier than referring to another person's, a finding which supports representational theory rather than the simulationist position. PMID- 9413799 TI - Cognitive deficits in parents from multiple-incidence autism families. AB - This study compares parents of two autistic children with parents of a Down syndrome (DS) proband, on tests of intelligence, reading and spelling, and executive function. Autism parents performed significantly worse than DS parents on performance IQ, a test of executive function, and some reading measures (e.g. passage comprehension and rapid automatized naming). These results suggest that cognitive deficits may be an expression of the underlying genetic liability for autism and that these characteristics may contribute to a more broadly defined autism phenotype. PMID- 9413800 TI - Planning and execution of action in children with and without developmental coordination disorder. AB - Ninety-five children from six English primary schools were identified on the basis of the Movement Assessment Battery for Children (Movement ABC) as having developmental coordination disorder (DCD) and, together with age- and ability matched controls, were given three tasks that involved proprioception in the control and discrimination of limb position, and two tasks that involved planning for end state comfort after a bar was grasped and turned. The children in the DCD group performed less well on the majority of the proprioceptive tasks, but did not differ from controls in planning of grip selection. There was an improvement in grip planning with age. The results are contrasted with research indicating that people with autism do have a difficulty with planning grip selection. PMID- 9413801 TI - Early intervention in adoptive families: supporting maternal sensitive responsiveness, infant-mother attachment, and infant competence. AB - Results from adoption studies suggest that adoptive families may experience special impediments with respect to the developmental progress and outcome of their children. Based on attachment theory, two early intervention programs were designed to support families in the Netherlands with an internationally adopted child. The intervention aimed at promoting maternal sensitive responsiveness, secure infant-mother attachment relationships, and infant exploratory competence. Ninety families with an interracially adopted infant (71 from Sri Lanka and 19 from Korea) were assigned to either a control group or one of two intervention groups. All of the children, 44 boys and 46 girls, were placed for adoption under the age of 5 months (M = 8 weeks). The first intervention group (N = 30) received a personal book, which focused on sensitive parenting. The second intervention group (N = 30) was provided with the same book as well as with three video feedback sessions at their home. The control group (N = 30) did not receive intervention. In the control group sensitive responsiveness and security of attachment were comparable to outcomes from normative samples. The least intensive program, the personal book, did not bring about change in mothers or infants. In contrast, intervention effects were established upon maternal sensitive responsiveness, infant competence, and infant-mother attachment in the group that received both the book and video feedback. PMID- 9413802 TI - Barriers to Treatment Participation Scale: evaluation and validation in the context of child outpatient treatment. AB - This study examined barriers that families experience during treatment and the role these barriers play in participation and completion of therapy. We developed the Barriers to Treatment Participation Scale and evaluated performance among children (N = 260, ages 3-13) and families referred for outpatient treatment. The results indicated that: (a) the scale yielded high levels of internal consistency; (b) the experience of barriers to participation, whether rated by parents or therapists, predicted higher rates of dropping out of treatment, fewer weeks in treatment, and higher rates of cancelled appointments and not showing up for sessions; (c) the perception of barriers was distinguishable from several family, parent, and child characteristics assessed at intake and the experience of critical life events during treatment; and (d) perceived barriers added significant information in predicting participation in treatment, over and above other characteristics that are already known to predict poor participation in treatment. Barriers associated with treatment participation can help identify cases at risk for dropping out and suggest targets for intervention to improve retention of families in treatment. PMID- 9413803 TI - Thyroid iodide transporter: local sequence homologies with thyroid autoantigens. AB - Here we show the existence of local amino acid (aa) sequence homologies between rat thyroid iodide transporter (Na+/l- symporter or NIS), whose gene was recently cloned, and known human thyroid autoantigens [thyroglobulin (Tg), thyroid peroxidase (TPO) and thyrotropin receptor (TSHR)] NIS sequences corresponding to the fourth (aa 264-282) and fifth extracellular loop (aa 386-414) are 15 to 40% identical and 30 to 60% similar to sequences corresponding to known or putative epitopes of Tg, TPO and TSHR. The sixth extracellular loop (aa 465-485) beared homology (44% identity, 52% similarity) only to a region of Tg which flanks one of its immunodominant domains. Sequences of thyroid autoantigens other than NIS shared homology, especially Tg and TPO. We conclude that in all likelihood NIS is an additional thyroid antigen, which shares common epitopes with the other thyroid autoantigens. Addendum: A study in abstract form appeared after submission of our paper finds experimental evidence for the antigenicity of two extracellular segments (aa 262-280 and 468-487) and of a portion of the intracellular C-terminus (aa 560-579). PMID- 9413804 TI - Different sensitivity to sodium valproate in healthy, non-tumoral and tumoral hyperprolactinemic subjects. AB - GABAergic drugs affect PRL secretion in both rat and man. Sodium valproate (SV) inhibits GABA transaminase so increasing the endogenous GABAergic tone. The aim of this study was to evaluate the effects of SV at low and high doses on PRL release in healthy subjects and hyperprolactinemic patients. Fifteen patients with prolactinomas, 8 patients with non-tumoral hyperprolactinemia and 10 healthy subjects were studied: in non consecutive days, all subjects received placebo and SV at the dose of 400 and 800 mg po. Serum PRL levels were assessed 30, 15 and 5 min before and every 30 min for 4 hours after administration. SV at the dose of 400 mg induced a significant decrease of serum PRL in healthy subjects (p < 0.05), whereas no effect was noted in both tumoral and non-tumoral hyperprolactinemia. The administration of 800 mg SV induced a significant decrease of PRL levels in healthy subjects and in patients with non-tumoral hyperprolactinemia (p < 0.05). Conversely, in prolactinomas a paradoxical increase of serum PRL concentration (p < 0.05) was observed 120 min after the administration of the drug. These data confirm the inhibitory activity of SV on PRL release in healthy subjects, and suggest the existence of a partial resistance to GABA in non-tumoral hyperprolactinemia. In prolactinomas, the paradoxical PRL increase after high dose of SV suggests the existence of a complete pituitary resistance to GABA. This finding might be explained by the appearance of the stimulatory effect of GABA at hypothalamic level that could have been unmasked by the lack of pituitary GABA effects on adenomatous lactotrophs. PMID- 9413805 TI - Menstrual cycle and ovary alterations in women with epilepsy on antiepileptic therapy. AB - Impaired reproductive function is thought to frequently affect women with epilepsy, mainly when seizures originate in the temporal lobe. In this study, we evaluated menstrual cycle features and assessed ovulation by determining luteal progesterone (Pg) levels in 101 consecutive women with epilepsy (36 with idiopathic generalized epilepsy -IGE; 65 with partial epilepsy -PE), aged between 16 and 50 years, treated with various antiepileptic drugs (AED). PE originated in the temporal lobe (TLE) in 40 subjects, in the frontal lobe in 13, in the parietal lobe in 2, while the origin of focal seizures remained undetermined in 10 patients. In all patients, menstrual and reproductive history, body mass index, hair distribution and hormonal pattern were assessed. Suprapubic ovary ultrasound (US) examination was carried out in 83 patients (28 with IGE, 55 with PE). Three patients with IGE and one with PE were amenorrheic. Oligomenorrhea occurred in 16 patients, polymenorrhea in 2. Changes in menstrual cyclicity were independent from epilepsy type (19.4% in IGE; 23.1% in PE) and from origin of focal discharges (22.5% of patients with TLE; 20.0% with origin in other brain areas). Luteal Pg levels remained below 2 ng/ml in 30 patients independently of epilepsy type. Corpus luteum dysfunction was combined with hyperandrogenism in 15 of these patients. In the other cases different alterations of hypothalamus pituitary-ovary axis were observed. Valproic acid blunted luteal Pg surge more frequently than other AED. Polycystic ovaries (PCO) were observed in 14 (16.9%) patients (21.0% with IGE: 14.5% with PE). These prevalences are not higher than those reported in the general population. Among PE patients, PCO was found in 1 case with undetermined focal origin and in 7 TLE cases, who also had ovary volume significantly larger than patients with seizures originating from the frontal or parietal lobe. Epileptic women exhibited an increased occurrence of multifollicular ovaries (MFO) found in 12 cases (14.4% vs 5% in the general population). However, no defined hormonal or clinical pictures were associated with this US alteration in most patients. These findings reappraise the impact of ovary alterations in women mainly affected by mild to moderate epilepsy, on differing AED regimens, with the exception of more frequent ovulatory dysfunction and PCO occurrence in patients taking VPA. PMID- 9413806 TI - Potential role of fluorine-18-deoxyglucose (FDG) positron emission tomography (PET) in the staging of primitive and recurrent medullary thyroid carcinoma. AB - We investigated 5 MTC patients, 3 preoperatively for staging purpose, and 2 after surgery, during the follow-up, because of the persistence of elevated serum tumoral markers. FDG PET results were compared with conventional radiologic (US, CT scan, MRI) and scintigraphic non-invasive techniques (99mTc-MIBI and 99mTc-MDP scans). In all the 3 patients preoperatively studied, PET, as well as the other imaging modalities, detected the primitive tumor and the loco-regional lymphnode metastases. Furthermore, in one case, PET was the only technique that revealed an additional localization to the lungs. One false negative result was recorded with PET, as well as with the conventional imaging, in a MTC patient with a MEN II syndrome and with some liver micrometastases, 2 to 5 mm sized, showed only at laparotomy. PET was the only method capable of early visualizing a mediastinal relapse of the tumor in one of the 2 patients studied during the follow-up. This patient was re-operated and serum calcitonin levels became undetectable. On the basis of our preliminary results on MTC, PET with FDG seems to be an accurate, non-invasive technique, for staging purpose before surgery, and, during the follow-up for visualizing tumoral spread in patients with increased serum tumoral markers. PMID- 9413807 TI - Sporadic amplification of the insulin receptor gene in human breast cancer. AB - Insulin receptor (IR) content is increased in most human breast carcinomas when compared to normal breast tissue. In the present study we investigated IR gene copy number by using both conventional DNA analysis (slot blot) and fluorescence in situ hybridization (FISH). Cultured human breast cell lines and primary breast carcinoma specimens were analyzed. In 6 breast cell lines in culture both techniques gave similar results: the relative IR copy number determined by FISH strongly correlated with slot blot results (r = 0.831), even if probes for different reference loci were used in the 2 methods. We find that in human breast cancer IR gene amplification is a sporadic event. It occurred in 1/5 cultured breast cancer cell lines (MDA-MB 231) and in 8/93 (8.6%) breast cancer specimens. In contrast an increased copy number of the entire chromosome 19 (which contains IR gene) was frequently observed in both breast cancer cell lines (100%) and breast cancer specimens (45%). When present, IR gene amplification always occurred at low level. These data indicate that IR gene amplification is an uncommon event in human breast carcinomas and that mechanisms other than gene amplification are responsible for IR protein overexpression in most human breast cancers. PMID- 9413808 TI - Use of cabergoline in the long-term treatment of hyperprolactinemic and acromegalic patients. AB - Cabergoline (Cab), a very potent and long-lasting dopaminergic compound, was administered to 26 women with pituitary microprolactinoma [mean serum PRL levels: 124.8 +/- 11.3 micrograms/l (+/- SE), range 62-300 micrograms/l] and 3 patients with GH-secreting pituitary adenoma (2 with associated PRL hypersecretion) for 12 and 24 months, respectively. In microprolactinomas, a stable normoprolactinemia was achieved in 96.1% of cases: in 13 women (50%) with the lowest dose of the drug (0.5 mg/week), and in other 12 patients (46.1%) with increasing doses up to 3 mg/week. All the oligomenorrheic/amenorrheic women, except one, restored regular and ovulatory menses. Two patients became pregnant. Pituitary abnormalities at high resolution-CT (HR-CT) scan disappeared in 13 of 19 patients (68.4%) after 12 months of therapy and this feature persisted in 8/13 cases (61.5%) 12 months after drug withdrawal. During Cab discontinuation (range: 3-60 months), mean serum PRL levels remained significantly lower than the basal ones. Six of 25 women are still without therapy. In 2 patients, normoprolactinemia persisted up to 38 and 60 months, respectively. Cab treatment was re-instituted in 13 patients because of the recurrence of hyperprolactinemia. Five patients were lost at follow up. In all the acromegalic patients, Cab (1-3 mg/week) normalized serum GH, IGF-I and PRL levels. A clear improvement in clinical symptoms was observed in all patients, but neuroradiological improvement in only one. Cab therapy was very well tolerated, as only seven patients complained of mild and transient side-effects and none had to stop treatment. In conclusion, Cab is an effective, safe, and well tolerated dopaminergic compound for the treatment of hyperprolactinemic disorders and the control of the clinical and hormonal features of dopamine-sensitive acromegalic patients. PMID- 9413809 TI - Long-term treatment with cabergoline, a new long-lasting ergoline derivate, in idiopathic or tumorous hyperprolactinaemia and outcome of drug-induced pregnancy. AB - Cabergoline (CAB), a new long-acting ergoline derivative, was shown to be very effective in reducing PRL levels in normal volunteers and in hyperprolactinemic patients. We evaluated the hormonal changes after discontinuation of long-term therapy with CAB as well as the safety of drug exposure during pregnancy both for mothers and babies. We therefore studied 48 patients (47 females and one male) with pathological hyperprolactinaemia (mean +/- SE, 117.2 +/- 15.2: median 73.2 micrograms/l), treated for 1-82 months (mean +/- SE, 28.3 +/- 3; median 18). After long-term treatment, CAB was withdrawn in 11 patients and PRL levels were persistently normal for almost 15 days and significantly lower (p < 0.05) than basal at 30, 45, 60, 90, 120 days. Three patients had normal PRL levels still at 45 days after treatment discontinuation. Nine patients became pregnant after 1-37 months (mean 12.4) of therapy. In two patients the pregnancy was interrupted spontaneously in one case and voluntarily in the other. In all but one patients after delivery or three-month breast feeding, PRL levels trended towards reduction. In two cases (one with microadenoma and one with idiopathic hyperprolactinaemia) PRL remained in the normal levels for 1-3 years after delivery. In conclusion CAB is able to inhibit plasma PRL levels for long time (up to 120 days) after withdrawal in patients with pathological hyperprolactinaemia treated with long-term therapy. PMID- 9413810 TI - Primary hyperparathyroidism-associated polyostotic fibrous dysplasia: absence of McCune-Albright syndrome mutations. AB - Several cases of sporadic primary hyperparathyroidism in association with fibrous dysplasia of the bone have been reported in the English literature. Since fibrous dysplasia is a major feature and hyperparathyroidism is occasionally found in the McCune-Albright syndrome, we hypothesized that such cases may represent a variant of this syndrome. A 28-year-old male had primary hyperparathyroidism associated with polyostotic fibrous dysplasia but no other manifestations of the McCune Albright syndrome. Genomic DNA samples from his parathyroid adenoma, dysplastic bone sample, and peripheral leukocytes were analyzed for the presence of activating mutations of the stimulating G protein alpha subunit gene (gsp). Allele-specific hybridization revealed the presence of normal sequences only, coding for arginine and glutamine at codons 201 (exon 8) and 227 (exon 9), respectively. Further, single strand conformational analysis of a 224 base pair fragment of exon 8 revealed no conformational aberrations. Furthermore, the sequences of a 164 base pair fragment of exon 8 and a 170 base pair fragment of exon 9 were normal. The results strongly suggest that gsp mutation is absent in affected and normal tissues in this patient and that the association of hyperparathyroidism and fibrous dysplasia may not represent a variant of the McCune-Albright syndrome. PMID- 9413811 TI - The measurement of urinary amino-terminal telopeptides of type I collagen to monitor bone resorption in patients with primary hyperparathyroidism. AB - This study was carried out in order to evaluate clinical usefulness of cross linked N-telopeptides (NTx) of type I collagen determination, in patients with primary hyperparathyroidism. Twenty-six consecutive patients (6 males and 20 females, aged 56.3 +/- 15.0, SD, yrs) with primary hyperparathyroidism were studied in basal conditions and, ten of them, after surgical cure of the disease. Cross-linked collagen peptides were measured by enzyme-linked immunosorbent assay and conventional markers of bone turnover according to standard procedures. Bone densitometry at the lumbar spine and proximal femur was performed using dual energy X-ray absorptiometry. Bone mineral density, was also assessed at the junction of the distal and middle third of the radius and at the ultradistal radius of the non-dominant arm by a dual photon densitometer. Mean urinary NTx values (194.2 +/- 121.9 pmoles bone collagen equivalents/mumoles creatinine) were significantly higher (p < 0.001) in respect to those found in normal subjects. The mean increase of Z score values of both serum tartrate resistant acid phosphatase activity (1.4 +/- 1.8) and the fasting hydroxyproline/creatinine ratio (1.45 +/- 2.0) was significantly lower (p < 0.02) in respect to that of NTx Z score values (3.3 +/- 3.3); the latter values were not significantly different than mean Z score values of serum osteocalcin (4.0 +/- 3.9), serum alkaline phosphatase activity (2.6 +/- 2.6) and urinary calcium/creatinine ratio (3.2 +/- 3.3). We found a significant inverse correlation between NTx values and both lumbar spine (p < 0.01) and ultradistal radius bone mineral density (p < 0.05); a modest inverse correlation was also observed between serum tartrate resistant acid phosphatase activity and lumbar spine bone mineral density (p < 0.04). Following successful adenoma removal, the percentage decrease of both NTx and hydroxyproline was similar in patients with increased bone turnover rate; major discrepancies were observed in patients with normal values of NTx, the telopeptide reduction being greater than that of hydroxyproline. Finally, in a hypercalcemic patient with metastatic parathyroid cancer, telopeptide excretion was shown to be more sensitive in respect to urinary hydroxyproline when evaluating the effects of antiresorptive therapy. Our results seem to indicate that amongst the markers with good sensitivity, NTx is the only one that is inversely related with bone mineral density at two different skeletal sites. This assay should therefore have a place in both the initial screening and medical follow-up of patients with this glandular disorder; in fact, in both situations an increased urinary excretion of this marker should warn about the possibility of hidden bone loss. PMID- 9413812 TI - Pituitary apoplexy of a gonadotroph adenoma following gonadotrophin releasing hormone agonist therapy for prostatic cancer. AB - Treatment of prostatic cancer with GnRH agonist is a medical alternative to surgical castration, although hyperstimulation of the tumor can occur. We describe an unusual unwanted effect of such a treatment which unmasked a clinically silent gonadotroph adenoma. A 62-year-old man developed after the first injection of leuprorelin-depot a sudden intracranial hypertension, which was related to apoplexy of an unknown pituitary adenoma. Its gonadotroph origin was recognized after surgery by immunocytochemistry. Retrospectively, the tumor was shown to secrete in vivo both FSH and LH when on therapy with the agonist, demonstrating the lack of desensitization. Testosterone levels were also markedly and sustainly high when on therapy, a particularly unwanted effect in prostatic cancer. As gonadotroph adenomas occur in men in the same age group as prostatic cancer, the question is raised whether hormonal testing and pituitary imaging should be performed before starting a therapy with GnRH agonist in men. PMID- 9413814 TI - Critical thinking--say what? PMID- 9413813 TI - Lactose intolerance following antithyroid drug medications. AB - We recently observed 2 lactase-deficient women with Graves' disease who consistently developed severe diarrhea after ingestion of thionamide (methimazole and propylthiouracil) tablets containing lactose as carrier. The strict temporal relationship between ingestion of lactose-containing tablets and appearance of intestinal symptoms, as well as the absence of side effects following ingestion of methimazole tablets without lactose as carrier, provided the clue for the diagnosis. To our knowledge, severe diarrhea resulting from carrier lactose has not been previously reported for antithyroid drugs, and should be considered in occasional cases of patients with gastrointestinal symptoms on thionamide therapy. PMID- 9413815 TI - Critical thinking in nursing: classroom tactics that work. AB - Critical thinking ability is one of the required outcomes of nursing education and content coverage is a focus of the past (National League for Nursing, 1996). As educators, we must attend to how we define critical thinking, what educational methods support its development, and how we can assure that students have achieved some acceptable level of critical thinking skill (Tanner, 1993). This article describes the emergent development of one critical thinking format in a Community Health Nursing course. Rather than approaching critical thinking from a theoretical perspective or focusing on one type of assignment or experience as a tool to foster critical thinking development in nursing courses, this article shares with the reader a complete package. Problems, pitfalls, new insights, and changes are shared as they developed through the teaching of a semester-long Community Health Nursing course. The authors hope their experiences give faculty ideas about how to infuse critical thinking into nursing curricula. PMID- 9413816 TI - A confirmatory factor analysis of the structure of tacit knowledge in nursing. AB - The purpose of this study was to examine the structure of tacit knowledge in the nursing profession. Using research in implicit learning and practical intelligence as a theoretical framework, a paper-and-pencil measure of tacit knowledge for nurses was developed and refined. Five models, each representing an established theory of traditional intelligence, were compared in terms of fit to the data. The results of the confirmatory factor analysis suggest that Model 3, a hierarchical model of intelligence, was the most plausible representation of tacit knowledge in nursing. The results indicate that the portion of managerial decision making learned implicitly on the job is mainly accounted for by managing tasks and others. Suggestions for nursing education include teaching effective strategies for managing tasks, such as handling increased workloads, establishing priorities, and delegating responsibility. PMID- 9413817 TI - Relationship between selected discourse strategies and student critical thinking. AB - This study investigated the relationship between use of selected discourse strategies and level of student critical thinking in nursing clinical post conferences. Selected discourse strategies included: (a) teacher high-level questions, (b) teacher elaboration of student ideas, (c) teacher probing questions, (d) student participation, and (e) student-to-student participation. The level of student critical thinking was defined as the quartile ranking of students (N = 57) on the Watson-Glaser Critical Thinking Appraisal summative measure. It was hypothesized that greater use of discourse strategies would be associated with high-levels of student critical thinking. All discourse strategies were significantly associated (p < .0001) to student quartile ranking for one taping session (Conference II). Further analysis, however, revealed ambiguous patterns of two-way associations. The more consistent findings suggested a conceptual relationship between less student talk and student-to student talk and high-levels of student critical thinking. PMID- 9413819 TI - Imagery: stimulating critical thinking by exploring mental models. AB - The use of guided imagery in the classroom or clinical setting has traditionally been directed at decreasing anxiety or increasing skill performance (Rodriguez, 1991; Stephens, 1992; Tuyn, 1994). This article describes an innovative use of imagery as a teaching strategy to unearth and possibly reframe, what Senge (1990) has identified as "mental models." The approach described differs from traditional uses of imagery related to decreasing anxiety and increasing skill performance. This approach is directed at increasing critical thinking and has offered new insights for both students and faculty. By using both guided imagery as a teaching strategy and Senge's concepts of mental models, educators can encourage students to become critical thinkers and what Senge calls "systems thinkers." Mental models are deeply ingrained, often unacknowledged, assumptions or images that individuals develop as a result of their life experiences. These images influence values, thoughts, and actions, albeit at times, unknowingly. These images, or mental models, also influence the ability to learn and translate learning into action. Lack of knowledge and awareness of one's mental models can be an obstacle to high-quality critical thinking. The concept of mental models has been best articulated by Senge (1990), who is known for his work in systems thinking. Senge believed that new insights are rarely followed with substantive action because the new learning, at some level, conflicts with deeply held, internal images. One way to remove this gap between insight and action is to examine mental models that may prohibit both insight and action. PMID- 9413818 TI - An action research study into the development of nurses as reflective practitioners. AB - This study adopted an action research approach in addressing the question: How could nurses be prepared to be reflective practitioners? The study took place among a group of registered nurses who enrolled in the first year of their degree studies at the Hong Kong Polytechnic University. The methods of data collection included observation, interview, students' written material, and teacher reflection. The experience of this study suggested that teachers and students should be partners in the promotion of reflective learning among students. As teachers reflected on their teaching arrangement and the progress of student learning, students could accordingly be offered appropriate guidance. Throughout the span of their studies, students learned to gradually develop different perspectives in viewing professional nursing practice. The reconceptualization of nursing practice helped students challenge taken-for-granted views and eventually reconstruct their conception of nursing. PMID- 9413820 TI - Evaluating critical thinking skills of baccalaureate nursing students. AB - This study evaluates the critical thinking skills of students enrolled in a baccalaureate nursing program, using the WGCTA, for the classes of 1993 through 1996. Scores were obtained at entry and at end of junior and senior years. The mean entry WGCTA score was 56 for all four classes; however, the 1995 and 1996 classes had significantly higher scores than the class of 1994. Critical thinking scores were higher at entry for older students and students who had completed another education degree; however, critical thinking scores were lower for students who had previous nursing experience. After adapting for age, previous degree, and nursing experience, no significant differences in the WGCTA scores from entry to end of junior and senior years emerged for the classes of 1993, 1994, 1995. Critical thinking skills have become the hallmark of education. The National Education Goal Panel has advocated for an increase in the ability to think critically, communicate effectively and solve problems (Banta, 1993). In turn, the nursing profession has incorporated these goals of higher education into its educational programs. The National League for Nursing (NLN) includes the measurement of critical thinking as a required outcome in the evaluation and accreditation of baccalaureate and higher degree programs in nursing. This critical thinking outcome must reflect the student's skill in analysis, reasoning, research, or decision making as these skills relate to the nursing discipline (National League for Nursing, 1992). To meet the NLN's critical thinking outcome criterion, nursing programs must have a method of evaluating this skill. Many programs use the Watson-Glaser Critical Thinking Appraisal (WGCTA), which is a standardized instrument. The College of Nursing at the University of Arkansas for Medical Sciences (UAMS) adopted this instrument to evaluate the critical thinking skills of students in the baccalaureate nursing program. PMID- 9413821 TI - Grading journals in clinical practice: a delicate issue. AB - Offering students opportunities to gain a strong sense of self, a positive professional image, and a chance to articulate their clinical practice is a challenge for nurse educators. Writing journals in clinical placements is one way in which students can create a dialogue with their teacher and reflect upon and explore their clinical experiences in the context in which these experiences occur. However, grading journals according to numerous predetermined criteria can sabotage the benefits and opportunities of writing journals. Judgment and control are two aspects of evaluation and subsequent grading that can sabotage the benefits. Limiting predetermined criteria and not assigning grades to students' journals are two answers to this delicate issue. To function as competent practitioners, nursing students must be able to meet standards of practice; they must achieve a strong sense of self and a positive professional image. Clinical placements offer students the opportunity to explore the experience of clinical nursing and the context in which these experiences unfold. As students acquire skills and explore the practice of nursing, they also face the reality that their instructors will award a judgment of worth to their efforts. This evaluation is necessary to determine whether students have met the required standards. Evaluation and subsequent grades, therefore, must be an integral part of the students' clinical experience. Writing journals is often used as a method of exploring experiences in clinical nursing. Journals are also used as a method of clinical evaluation. Assigning a grade to student journals has a detrimental effect on the purpose of the assignment. An emphasis on exploring the purpose of writing journals and an analysis of the impact that grading has on this exercise will expose the incompatibility between writing and grading journals. PMID- 9413822 TI - Mechanical impedance of layered tissue. AB - The human body is a medium which consists of various tissues such as skin, fat, muscle and bone. Each of the tissues has their own biomechanical properties. We have measured biomechanical impedances by applying a random vibration (30-1000 Hz) to the layered model of human tissues to study the occurring mechanism of impedances measured at the skin surface. The data showed that the top tissue layer and the underlying layer both contribute to the impedance depending on the thickness of the top layer. The contribution of the underlying layer was clearer over the frequency range from 30 to 400 Hz. Quantitatively we found the following: The impedance measured at the surface was roughly expressed as the model which is connected in series by the impedances of the top and underlying tissues. The contribution of the underlying tissue decreased according to the increase of the thickness of the top tissue, and disappeared over a certain thickness (18 mm in this paper). PMID- 9413823 TI - Non-invasive assessment of systemic elastic behaviour in hypertensive patients: analysis of possible determinants. AB - Knowledge about the viscoelastic behaviour of the arterial wall has been proved to have physiological importance and clinical usage. Our purpose was to study the changes of the systemic arterial wall's elastic properties non-invasively, in patients with established essential and with borderline hypertension, and to evaluate its possible determinants. Three groups of normotensive, borderline and established essential hypertensive patients were evaluated. Arterial pulse wave velocity (PWV) was measured and arterial compliance (Cm) was derived in all patients. Pulse wave velocity was obtained from the pressure values of digitized carotid and radial arteries. Arterial compliance (Cm = dD/dP with P pressure and D diameter) was calculated using a formula derived from the Bramwell and Hill equation: Cm = (1,334 x D)/(2 rho x PWV2), where for D humeral diameter was used as measured by high resolution echograph, and rho is the blood density (rho = 1.06). Pulse wave velocity was significantly higher in established essential hypertensive patients with respect to normotensive patients (p < 0.05). Arterial compliance was significantly diminished in established and in borderline hypertensive patients with respect to normotensive patients (p < 0.05), which implies early alterations in hypertensive cardiovascular disease. Multiple regression analysis of the cofactors showed that age and diastolic pressure are independent determinants of Cm. Impairment of the arterial wall's intrinsic elastic properties was demonstrated in established essential hypertension, independent of age and diastolic pressure. PMID- 9413824 TI - Guinea pig auditory nerve response triggered by a high density electrode array. AB - The electrically evoked potential in the auditory nerve is measured when the cochlear is stimulated with a high density electrode array whose microcontacts (20 x 160 microns) are placed close to the nerve cells. Threshold range from 8 to 35 microA with the stimulating electrodes near the spiral ganglion cells. A multi pole technique for restricting the spread of current with electrical stimulation of the cochlea is tested using neural recording in deafened guinea pigs. The ground based quadrupolar electrode driving configuration has thresholds for neural activation only slightly greater than monopolar stimulation when the electrode contacts are placed less than 50 microns from the neurons. During simultaneous stimulation the monopole and the ground based quadrupolar modes tend to generate similar growth functions (magnitude and latency). The electrode interactions are generally factorial, which means that the algebraic sum of the responses (magnitude growth functions) produced by 2 distinct electrodes is less than when the same 2 electrodes are stimulated simultaneously. PMID- 9413825 TI - A new computerized control unit for small bone surgical instruments. AB - The SmartDrive console represents an important advance in small bone surgery because it monitors and coordinates the operation of its handpieces. The SmartDrive console has the following unique features: 1) a handpiece recognition system; 2) an instrument speed display; 3) a handpiece display and monitoring system; 4) a torque instrument control system; 5) a temperature monitoring system; 6) and an irrigation system. Mechanical performance studies have been undertaken that have validated the accuracy of the monitoring systems of the consoles. The consoles provided reliable recordings of the rotational speeds of their hi-speed drills. The MicroAire console automatically shuts off its power as the temperature increased to 110 degrees F (43 degrees C). In contrast, the Stryker Command 2 console has a limited monitoring system that can not alter the operation of the handpieces. PMID- 9413826 TI - Immunoregulatory activities of L-4-oxalysine: an in vitro study. AB - The present study was undertaken by in vitro experiments to determine the effect of L-4-oxalysine (OXL), a new natural product in China, on immunological responses. The immunological parameters evaluated were one-way mixed lymphocyte reaction and interleukin-6 activity stimulated by lipopolysaccharide. The results showed that the immunological functions of spleen cells from normal mice were significantly suppressed by in vitro treatment of OXL. However, OXL markedly enhanced the immunological responses of spleen cells from mice bearing hepatoma 22 tumor. It was indicated that OXL exerted obvious immunoregulatory activities. PMID- 9413827 TI - The ionic mechanisms of early after depolarization in mouse ventricular myocytes: the role of IK1. AB - The occurrence of early after depolarization (EAD) in single mouse ventricular myocytes was observed and its ionic mechanisms were studied using the patch clamp technique. Under treatment with perfusion of Tyrode's solution containing 3 mM KCl and 3 mM CsCl, 3/6 cases exhibited EAD, while with 3 mM KCl or 3 mM CsCl alone, EAD was not induced. The background steady-state current-voltage (I-V) curves of the myocytes showed no negative slope, i.e., the slope in the range of 50 mV positive to the reversal potential was virtually flat and stayed at a low current level. Under perfusion of 3 mM KCl and 3 mM CsCl, the outward current in the above region decreased nearly to 0: in the myocytes which exhibited EAD, a net inward current (crossover) was displayed in the same region, which was abolished by 10 microM TTX and 10 microM nifedipine. The results of whole-cell inward rectifier current I-V curves were similar to the above background steady state I-V curves. In mouse ventricular myocytes, transient outward current was very strong with a peak current density of 63 +/- 19 pA/pF, whereas low K+ and Cs+ had no significant effect. 11/30 cases showed obvious delayed rectifier current, but the tail current recorded by envelope method was relatively weak (1.19 +/- 0.35 pA/pF) and insensitive to CsCl or changing of the KCl concentration. The results suggest that under treatment with low K+ and Cs+, the inhibition of inward rectifier current is the basis of the formation of second plateau, while Na and Ca currents contribute to the generation of triggered bursts. PMID- 9413828 TI - Administration of a nitric oxide donor does not affect hypotension induced by 35 GHz microwave heating. AB - Sustained whole body exposure to 35-GHz radiofrequency radiation produces localized hyperthermia and hypotension, leading to circulatory failure and death. We previously demonstrated that pressor responses to nitric oxide (NO) synthesis inhibition are reduced following 35-GHz microwave (MMW) heating, implying that NO levels might also be reduced. This study therefore sought to determine whether administration of S-nitroso-N-acetylpenicillamine (SNAP), a NO donor, influences MMW-induced hypotension in ketamine-anesthetized rats. First, rats were exposed to MMW until mean arterial pressure (MAP) fell to 75 mmHg. MMW exposure was then discontinued and either SNAP (300 micrograms/kg/h) or saline was infused. SNAP infusion affected neither the hypotension nor the survival time following MMW exposure. In a second protocol, SNAP (300 micrograms/kg/h) or saline was infused prior to and throughout MMW exposure, which was continued until death. SNAP infusion did not alter either the onset or the magnitude of terminal hypotension. Therefore, we conclude that exogenous NO does not affect cardiovascular responses to 35-GHz MMW heating. PMID- 9413829 TI - The role of adrenal hormones in the activation of tryptophan 2,3-dioxygenase by nicotinic acid in rat liver. AB - In this study, our previous finding that nicotinic acid activates tryptophan 2,3 dioxygenase as strongly as tryptophan was investigated in further detail. This study focused on the role of the adrenals in the activation process. Adrenalectomy abolished the activation due to nicotinic acid, but not the activation caused by tryptophan. The role of corticoids and/or adrenomedullary hormones in the enzyme activation was studied, by supplementing these hormones in adrenalectomized rats using minipumps implanted under the skin. The results showed that the enhanced activity of tryptophan 2,3-dioxygenase caused by nicotinic acid was partly restored by adrenaline following adrenalectomy but not by corticosterone supplementation. The results were supported by further experiments in which the rats were treated with adrenaline or corticosterone intraperitoneally before nicotinic acid administration. The conclusion that adrenaline participates in the regulation of tryptophan 2,3-dioxygenase should promote further study to determine whether adrenaline is a general modulator of this enzyme. This experimental model generated new information on the activation mechanism of tryptophan 2,3-dioxygenase by nicotinic acid. PMID- 9413830 TI - Intrahippocampal injections of the beta-amyloid 1-28 fragment induces behavioral deficits in rats. AB - beta-amyloid (beta A) deposition is a key event in the etiopathogenesis of Alzheimer's disease (AD), contributing to neuronal degeneration and cognitive impairment in AD patients. Both neurotrophic and neurotoxic actions of beta A have been demonstrated in experimental conditions. In order to further characterize the effects of brain beta A deposits on behavioral processes, we evaluated psychomotor activity (PMA), psychomotor coordination (PMC) and learning in a passive avoidance task (PAL) in rats with unilateral or bilateral 2 microliters injections of beta-amyloid (1-28) protein (beta A; 1.5 nmol/microliter) or vehicle (water; W) into the hippocampus, 1 and 4 weeks after neurosurgery. The extent of neuronal loss in the lateral blade of the gyrus dentatus (LBGD) and the area percentage occupied by APP immunoreactivity in neurons of the CA3c subfield of the hippocampus were also measured in animals with unilateral beta A implants. PMA levels were similar in water- and beta A injected animals 1 and 4 weeks after recovery. As compared to water-injected rats, beta A animals showed reduced PMC values 1 week, but not 4 weeks, after injections. beta A also impaired learning acquisition in a passive avoidance task, reducing the number of avoidances and mean latency per trial at both 1 and 4 weeks postsurgery in rats with unilateral or bilateral beta A implants. The extent of neuronal loss in the LBGD) was not different in rats receiving water or beta A injections. Hippocampal APP expression tended to increase in beta A implanted rats and showed a negative correlation with cognitive performance at the 4-week period. According to these results it seems that beta A implants into the hippocampus reduce psychomotor coordination performance in a transient manner, with no effect on psychomotor activity, and induce durable learning impairment in rats, and that changes in cognitive performance correlate with histochemical parameters such as APP expression. In conclusion, the present results contribute to a better understanding of beta A-induced behavioral alterations and to the identification of potential molecular mechanisms involved in cognitive dysfunctions in this animal model of neurodegeneration. PMID- 9413831 TI - Dopaminergic modulation of lithium/pilocarpine-induced status epilepticus in rats. AB - The effects of selective dopaminergic agonists and antagonists on epileptic activity were tested in rats using the lithium/pilocarpine seizure model. Systemic administration of the D1 agonist SKF-38,393 reduced the latency of onset of forelimb clonus with rearing, whereas the D1 antagonist SCH-23,390 and the D2 agonist B-HT 920 prevented the convulsive activity in a dose-dependent manner. Mixed agonists like apomorphine offered partial protection. Haloperidol (D1, D2 blocker, with antimuscarinic property) reduced the threshold for convulsions. The effects of SKF-38,393 and B-HT 920 could be partially blocked by pretreating the rats with SCH-23,390 and sulpiride, respectively. Neither D1 nor D2 antagonists could alter the limbic stereotypies induced by lithium/pilocarpine. These results indicate that dopamine receptor subtypes exert opposite effects on the regulation of convulsive activity. Lipid peroxidation levels (MDA formed) in rat brain homogenates were found to be concomitant with the development of epileptiform activity. PMID- 9413832 TI - Effects of feeding egg yolk on the serum lipid levels in rabbits. AB - Healthy, nonobese, young rabbits developed hypercholesterolemia following daily intake of fresh egg yolk for 8 weeks. On a quantitative basis, this diet had a profound effect on serum cholesterol level which rose to 15 or 30 times the baseline value depending on whether the test group consumed the yolk content of one or two eggs during the study. Similar drastic changes observed in low density lipoprotein cholesterol levels in these test animals make this a useful method for defining the early pathological changes of atherosclerosis in man. In addition, the present study provides direct evidence of the deleterious effect of hen's egg yolk on the serum cholesterol level in an appropriate and easily reproducible animal model. PMID- 9413833 TI - The possible role of adrenergic component in ischemic preconditioning. AB - The role of adrenergic mechanism in the cardioprotective effect of ischemic preconditioning against ischemia-reperfusion induced injury in in vivo dog heart and isolated rat heart was investigated. Anesthetized dogs were subjected to LAD coronary artery ligation for 60 min followed by reperfusion for 4 h. Preconditioning protocol was 5 min of ischemia followed by reperfusion for 10 min. Rat hearts were subjected to global ischemia for 30 min followed by reperfusion for 30 min. Preconditioning protocol was 5 min global ischemia followed by reperfusion for 5 min repeated four times. Infarct size, electrocardiographic changes and release of LDH were estimated to assess the extent of cardiac injury. Preconditioning reduced the infarct size, ST segment elevation and prevented the loss of R wave. Prazosin attenuated the cardioprotective effect of preconditioning in dog. Preconditioning conferred protection against ischemia-reperfusion induced cardiac injury and reperfusion induced arrhythmias in isolated rat heart. Reserpine pretreatment attenuated this protective effect of preconditioning on reperfusion-induced arrhythmias. These observations suggest the involvement of adrenergic mechanism in the cardioprotective and antiarrhythmic effect of ischemic preconditioning in dog and rat species respectively. PMID- 9413834 TI - Solubility of calcium salts and their effect on osteoporosis. PMID- 9413835 TI - Are exocytosis mechanisms neurotransmitter specific? AB - Neurotransmission is a multistage regulated process in which a variety of active molecules contained in vesicles are liberated in response to specific stimuli from different types of neurone or related cells. This includes the release of fast neurotransmitters such as amino acids and acetylcholine from central and peripheral synapses, but also that of relatively slow-acting polypeptides from central and peripheral neurones or neuroendocrine cells. Considerable progress has been made over recent years in the understanding at a molecular level of the mechanism of regulated exocytosis, a crucial phase in this phenomenon. The currently proposed overall mechanism, which incorporates the "SNARE" hypothesis for vesicle-membrane docking and fusion, is based on data from experimental models ranging from brain synaptosomes to mast cells. Since the kinetics of the models studied and the physiological effects of the neurotransmitters implicated vary so much, it is pertinent to question whether a general mechanism can be proposed from such experimental data. This review examines known differences in putative exocytotic mechanisms for the various systems studied and attempts to relate these to the nature of the active substances released. Differences exist in each step of the exocytosis process and include the channel through which Ca2+ enters to trigger it or the internal Ca2- source, the type of vesicle in which the transmitter is packaged, the way vesicles are translocated to the surface membrane or how they dock and fuse with it. Major differences have been reported in release mechanisms of different types of vesicle, but minor differences also exist within the same vesicle class. Thus small synaptic vesicles and large dense core vesicles are translocated by distinct processes and the Ca2+ channels, Ca2+ sensors and docking proteins involved in other steps are not identical in all neuronal phenotypes. It may be concluded that each of these differences has evolved to accommodate the different physiological requirements of the neuromodulator released. PMID- 9413836 TI - Are mechanisms of exocytosis neurotransmitter specific? AB - In their review, Langley and Grant (1997) investigate the question whether mechanisms of exocytosis are neurotransmitter specific. There is now much evidence that the mechanisms governing the exocytosis of the two principal storage organelles--granules (large dense core vesicles) and electron-lucent vesicles--differ. But much less is known concerning potential differences in the release mechanisms of electron-lucent vesicles that store different types of fast neurotransmitters or of granules in different types of neurons. It is an open question whether there is a unifying control mechanism for the exocytosis of, for example, a peptide-containing granule of a glutamatergic neuron, a chromaffin granule, a noradrenergic granule or a granule from a neurosecretory neuron in the pituitary. The small electron-lucent synaptic vesicles of various kind apparently share common molecular components of regulated release. They carry the calcium sensor synaptotagmin, small GTP-binding proteins of the rab3 group or the v-SNARE synaptobrevin. Nevertheless, there may be differences in the regulatory mechanisms. This concerns the type of calcium channel involved or the absence of some of the presynaptic molecules such as rab3a, synapsin I or the t-SNAREs SNAP 25 or syntaxin from distinct types of neurons or sensory cells. PMID- 9413837 TI - An inevitable gate for quantal acetylcholine release. PMID- 9413838 TI - GABA augments basal and electrically stimulated 3H-norepinephrine release in hypothalamic, preoptic area and cortical slices of female rats. AB - These studies examined the regulation by GABA of norepinephrine release from hypothalamus, preoptic area and frontal cortex. Using superfused brain slices from female rats, we show that 100 microM GABA enhances both basal and electrically stimulated release of 3H-norepinephrine in all three brain regions. The GABAA agonist muscimol (100 microM) significantly augments 3H-norepinephrine release, but it is somewhat less effective than GABA. The GABAB agonist baclofen has little or no effect on basal 3H-norepinephrine efflux. GABA also augments both the magnitude and duration of electrically evoked 3H-norepinephrine release in slices from all three brain regions. GABA facilitation of electrically stimulated 3H-norepinephrine release is mediated through GABAA receptors as evidenced by its blockade by 10 microM bicuculline, a GABAA antagonist, but not by 200 microM 2-OH-saclofen, a GABAB antagonist. These data show that the inhibitory amino acid neurotransmitter GABA enhances both basal and evoked release of 3H-norepinephrine in brain slices from female rats. These effects are predominantly mediated by GABAA receptors. GABA modulation of hypothalamic norepinephrine release may play a role in the regulation of gonadotropin secretion and reproductive behaviors such as lordosis. PMID- 9413839 TI - Binding of the benzodiazepine ligand [3H]-Ro 15-1788 to brain membrane of the saltwater fish Mullus surmuletus. AB - The distribution and the pharmacological properties of the binding of the benzodiazepine receptor antagonist [3H]-Ro 15-1788 (8-fluoro-3-carboethoxy-5,6 dihydro-5-methyl-6-oxo-4H imidazol [1,5-a] 1,4 benzodiazepine) were compared in some brain membranes of the saltwater teleost fish, Mullus surmuletus: only a single population of [3H]-Ro 15-1788 binding sites was detected. The binding was saturable and reversible with a high affinity, revealing a significant population of binding sites (Kd value of 2.1 +/- 0.2 nM and Bmax value of 1400-900 fmol mg-1 of protein, depending on fish length). The highest concentration of benzodiazepine recognition sites labelled with [3H]-Ro 15-1788 was present in the optic lobe and the olfactory bulb and the lowest concentration was found in the medulla oblongata, cerebellum and spinal cord. In order to explore behavioural selectivity as a consequence of multiple receptor subtypes, six benzodiazepine receptor ligands, flunitrazepam (5-(2-fluoro-phenyl)-1,3-dihydro-1-methyl-7-nitro 2H-1, 4-benzodiazepin-2-one), alpidem, (N,N-dipropyl-6-chloro-2-(4 chlorophenyl)imidazo [1,2-a] pyridine-3- acetamide) zolpidem {N,N,6, trimethyl-2 (4-methyl-phenyl) imidazo [1,2-a] pyridine-3-acetamide hemitartrate}, methyl beta carboline-3-carboxylate (beta CCM), Ro 15-1788 and Ro 5-4864 (4'-chlorodiazepam), were tested in vitro by binding of [3H]-Ro 15-1788 to membrane preparations from various brain areas of Mullus surmuletus, Displacement studies showed a similar rank order of efficacy of various unlabelled ligands. In all regions of the brain and in the spinal cord. GABA potentiate [3H]-flunitrazepam binding in a similar order, suggesting that the BDZ recognition sites are part of the GABAA receptor structure. These results suggest that central-type benzodiazepine receptors are present in one class of benzodiazepine binding sites in the saltwater teleost fish brain of Mullus surmuletus (type I-like). Here we report initial evidence of homogeneity of subtypes of central benzodiazepine receptors in the spinal cord of the saltwater teleost fish. Mullus surmuletus. PMID- 9413840 TI - Influence of dietary fat on the activities of subcellular membrane-bound enzymes from different regions of rat brain. AB - The effect of different dietary fats with varying degrees of unsaturation and essential fatty acid composition, which are commonly consumed in India, on the activity of some important membrane-bound enzymes was assessed in different brain regions of rat. Four groups of male CFY weanling rats were fed nutritionally adequate diets containing groundnut, coconut, safflower or mustard oil as fat source at 20% level for 16 weeks. The synaptosomal, microsomal and myelin membranes were prepared from three brain regions, viz., cerebrum, cerebellum and brain stem from each group. The activities of Na+, K(+)-ATPase, Mg(2+)-ATPase and acetylcholinesterase were assayed and the fatty acid composition was determined in these subcellular membrane fractions. The safflower oil-fed group showed higher Na+, K(+)-ATPase activity in most membrane fractions than the coconut or mustard oil-fed groups. The Mg(2+)-ATPase activity was found to be similar amongst all groups in all the brain regions. The synaptosomal acetylcholinesterase activity was distinctly higher in coconut and groundnut oil fed groups when compared to safflower or mustard oil consuming groups. Alterations in the activities of these subcellular membrane-bound enzymes are expected to exert a significant impact on the electrophysiological and metabolic functions of brain. Results of the present study show that depending on the nature of dietary fat the fatty acid composition of subcellular membranes is altered, which in turn could regulate the activity of membrane-bound enzymes that are vital for brain function. PMID- 9413841 TI - Inhibitory cholinergic control of endogenous GABA release from electrically stimulated cortical slices and K(+)-depolarized synaptosomes. AB - In the present study we characterize the optimal experimental conditions under which to investigate the cholinergic regulation of endogenous electrically evoked gamma-aminobutyric acid (GABA) release from guinea pig cortical slices. Superfusion with the neuronal GABA reuptake inhibitor, SKF89976A (10 microM) caused cortical GABA release to be linearly correlated with the frequency of electrical stimulation (5, 10, 20 Hz). Electrically evoked GABA release (10 Hz) was tetrodotoxin-sensitive and Ca(2+)-dependent and was under GABAB autoreceptor control. Under these experimental conditions, acetylcholine (0.1-10 microM) and physostigmine (30 microM) decreased the electrically evoked GABA release while the M2 receptor antagonist AFDX-116 (0.01-0.1 microM) counteracted these effects. Similar results were also observed in a cortical synaptosomal preparation stimulated with K+ (10 mM). These findings demonstrate an inhibitory cholinergic regulation of electrically evoked GABA release via M2 receptors located on cortical GABAergic terminals. PMID- 9413842 TI - Localization and characterization of stannin: relationship to cellular sensitivity to organotin compounds. AB - The cDNA encoding the protein stannin was isolated previously via subtractive hybridization, using differential expression after trimethyltin (TMT) intoxication, as a basis for isolating mRNA which may be expressed in TMT sensitive cells. Initial characterization revealed a novel gene product which was differentially expressed in several tissues sensitive to TMT. In the current study, biochemical and molecular techniques were used to quantitate stannin expression at the cellular and subcellular levels. Northern blot analysis showed that the stannin 3.0 kb mRNA transcript was present, in decreasing amounts, in: spleen, hippocampus, neocortex, cerebellum, striatum, midbrain, kidney and lung. Liver, heart, skeletal muscle and testis showed no detectable expression of stannin mRNA. Immunoblot analysis using antipeptide antisera raised against stannin indicated a high level of expression in spleen, followed by brain and kidney. Stannin mRNA was present during early brain development and consolidated by post-natal day (PND) 20 to patterns and levels seen in adults. In situ hybridization studies showed widespread neuronal expression of stannin mRNA at PND 1, which shifted to a restricted pattern of expression in specific regions by PND 20. Stannin was partially purified from rodent brain and spleen using cation exchange, sizing and hydrophobic interaction chromatography. It behaved as a monomer throughout purification. Stannin was also expressed in a baculovirus system, using a series of constructs containing the entire cDNA, 1.0 kb of DNA flanking the open reading frame, and a 400 bp open reading frame minimal construct. While all constructs expressed stannin, the best expression was seen with the entire cDNA. Based on current findings, we suggest that stannin expression is necessary but not sufficient for TMT toxicity. PMID- 9413843 TI - Post-ischemic recovery of acetylcholine release in vitro: influence of different excitatory amino acid receptor subtype antagonists. AB - The release of endogenous acetylcholine was measured in electrically (5-20 Hz) stimulated guinea pig cerebral cortex and caudate nucleus slices under ischemic (hypoxic and glucose-free) conditions. Ischemia reduced acetylcholine release by 40-90%; the inhibition depended on the duration of ischemia (10-30 min) while the extent of post-ischemic recovery was inversely related to it. Caudate nucleus slices displayed a higher sensitivity to ischemia than did cortical slices. To test the effects of excitatory amino acid receptor antagonists on the ischemia induced reduction of acetylcoline release and on its post-ischemic recovery, the following drugs were used: 5-methyl-10,11-dihydro-5-H-dibenzo-[a,b]-cyclohepten 5,10-imine (MK-801,-a blocker of the N-methyl-D-aspartate [NMDA] receptor-linked channel), 7-chloro-kynurenic acid (7-Cl-KYN) and (E)-3 [2(phenylcarbamoyl)ethenyl]-4,6-dichloroindole-2-carboxylic acid sodium salt (GV150526A, blockers of the glycine site of the NMDA receptor), eliprodil, (an antagonist at the polyamine site of the NMDA receptor), and 6-cyano- 7-nitro quinoxalin-2,3-dione (CNQX, a D,L-alpha-amino-3-hydroxy-5-methyl-4-isoxalone propionic acid [AMPA] receptor antagonist). These did not modify the time-course and the extent of ischemia-induced inhibition but improved post-ischemic recovery in a concentration dependent manner. GV 150526A and CNQX appeared to be more effective in the cerebral cortex. Only eliprodil was devoid of any effect in both areas. The evaluation of acetylcholine release from brain slices represents a suitable in vitro model to quantify the effectiveness of drugs in favouring recovery from the cholinergic presynaptic failure induced by ischemic conditions. The different effects of the excitatory amino acid receptor antagonists cited above, depending on the brain areas considered and the receptor subtypes involved, may be of interest in view of their therapeutic potential. PMID- 9413844 TI - Stable interaction between G-actin and neurofilament light subunit in dopaminergic neurons. AB - Excessive accumulation of neurofilaments in the cell bodies and proximal axons of motor neurons is a major pathological hallmark of motor neuron diseases. In this communication we provide evidence that the neurofilament light subunit (68 kDa) and G-actin are capable of forming a stable interaction. Cytochalasin B, a cytoskeleton disrupting agent that interrupts actin-based microfilaments, caused aggregation of neurofilaments in cultured mesencephalic dopaminergic neurons, suggesting a possible interaction between neurofilaments and actin; which was tested further by using crosslinking reaction and affinity chromatography techniques. In the cross-linking experiment, G-actin interacted with individual neurofilament subunits and covalently cross-linked with disuccinimidyl suberate, a homobifunctional cross-linking reagent. Furthermore, G-actin was extensively cross-linked to the light neurofilament subunit with this reagent. The other two neurofilament subunits showed no cross-linking to G-actin. Moreover, neurofilament subunits were retained on a G-actin coupled affinity column and were eluted from this column by increasing salt concentration. All three neurofilament subunits became bound to the G-actin affinity column. However, a portion of the 160 and 200 kDa neurofilament subunits did not bind to the column, and the remainder of these two subunits eluted prior to the 68 kDa subunit, suggesting that the light subunit exhibited the highest affinity for G-actin. Moreover, neurofilaments demonstrated little or no binding to F-actin coupled affinity columns. The phosphorylation of neurofilament proteins with protein kinase C reduced its cross-linking to G-actin. The results of these studies are interpreted to suggest that the interaction between neurofilaments and actin, regulated by neurofilament phosphorylation, may play a role in maintaining the structure and hence the function of dopaminergic neurons in culture. PMID- 9413845 TI - Hypoosmotic shock activates Ca2+ channels in isolated nerve terminals. AB - Influence of hypotonic swelling on Ca2+ (45Ca2+) uptake in rat brain synaptosomes was studied. A decrease in medium osmolality from 310 to 260-180 mOsm led to a progressive stimulation of 45Ca2+ accumulation. The effect was blocked by verapamil (IC50 = 5 microM), CoCl2 (IC50 = 58 microM) and retained at a fixed concentration of external sodium indicating the involvement of Ca2+ channels rather than Na+/Ca2+ exchange in swelling-induced Ca2+ influx. The populations of calcium channels observed in hypoosmotic and depolarizing conditions are different in three aspects: (i) kinetics of 45Ca2+ entry; (ii) insensitivity to dihydropyridines and omega-conotoxin GVIA; (iii) insensitivity to preliminary depolarization by high potassium. The effects of swelling and depolarization on Ca2+ uptake were additive. No change in membrane potential monitored with diS-C3 (5) was recorded during synaptosome hypotonic swelling. The results suggest the existence in synaptosomal plasma membrane of volume-dependent calcium-permeable channels with properties distinct from those of the voltage-dependent calcium channels. Activation of these channels may constitute an early event in volume regulation of nerve terminals in anisoosmotic conditions. PMID- 9413846 TI - N-methyl-norsalsolinol, an endogenous neurotoxin, inhibits tyrosine hydroxylase activity in the rat brain nucleus accumbens in vitro. AB - As a compound of structural analogy with MPTP, N-methyl-norsalsolinol (2-methyl 6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline; 2-MDTIQ) was recently identified in the brain and cerebrospinal fluid of patients with Parkinson's disease. As 2 MDTIQ cannot pass the blood-brain barrier, endogenous formation is suggested. Previous studies of the dopamine metabolism in Parkinson's disease have demonstrated an increased dopamine turnover in the presence of 2-MDTIQ. In the present study, we investigated the effect of 2-MDTIQ on tyrosine hydroxylase [L tyrosine, tetrahydropteridine, oxygen: oxidoreductase (3-hydroxylating). EC 1.14.16.2; TH] activity in vitro using homogenated tissue of the rat nucleus accumbens as enzyme source. Basal TH activity was 20.1 +/- 5.9 pmol L-3,4 dihydroxyphenylalanine (L-DOPA)/min/mg protein. 2-MDTIQ non-competitively inhibited basal TH activity with an IC50 of 10 microM. After addition of 0.1 mM 2 MDTIQ, enzyme activity was nearly completely blocked. These results indicate that the endogenous formation of 2-MDTIQ in consequence of an impaired dopamine metabolism may in turn lead to a decrease in dopamine synthesis. Thus, 2-MDTIQ is suggested not only to represent an endogenous marker of Parkinson's disease, but also to support changes in the transmitter synthesis of dopaminergic neurons. Since previous investigations have moreover demonstrated a cytotoxic potential of 2-MDTIQ, these findings require special attention. 2-MDTIQ may represent an essential factor in the degenerative process of Parkinson's disease. PMID- 9413847 TI - Lipoproteins and coagulation. AB - A broad range of molecular and cellular interactions contribute to various pathophysiological alterations in haemostasis. Recent studies have shown strong links between lipoproteins and coagulation factors. Findings suggest that lipoproteins play an important role in the fibrinolytic and thrombogenic mechanisms that influence the risks of patients in acute coronary syndromes. We will examine specific aspects of lipoproteins with reference to the effects of hyperlipidaemia on endothelial dysfunction and haemostasis, and its relevance in the patient presenting for revascularization. PMID- 9413848 TI - Clinical evaluation of six hollow-fibre membrane oxygenators. AB - In a clinical evaluation to study the performance of hollow-fibre membrane oxygenators, we compared the gas exchange characteristics and the production of plasma free haemoglobin for different oxygenators. In this study, the data of the Univox, Cobe-Optima, Capiox-SX, Affinity, Safe II and Sarns Turbo 440 oxygenators were evaluated during cardiac surgery in comparable patient groups. Thirteen patients scheduled for elective surgery were enrolled in each group. In all groups, cardiopulmonary bypass (CPB) was conducted using pulsatile blood flow during fibrillation and the period in which the aorta was crossclamped. Arterial and venous blood gases were determined from which oxygen transfer, carbon dioxide transfer, oxygen gradient, shunt fraction and diffusing capacity were calculated. Blood samples were taken 5, 30, 60 and 90 min after beginning CPB. As a result of our measurements we found that the gas exchange capacities of all six oxygenators are within clinically acceptable limits, although the data considering oxygen and carbon dioxide transfer showed a significantly higher capacity for the Sarns Turbo 440 oxygenator (p < 0.05) in comparison with the other oxygenators. Plasma free haemoglobin was, however, significantly higher in the Univox and Sarns Turbo 440 groups (p < 0.005). This difference was already present after 30 min of bypass and increased with time, and is considered as a negative aspect, in relation to optimal patient care. PMID- 9413849 TI - Biocompatibility of three different membrane oxygenators: effects on complement, neutrophil and monocyte activation. AB - The inflammatory reaction of extracorporeal circuits can be assessed by measuring complement activation and the release of activation markers of leucocytes. The purpose of this study was to compare three commercially available membrane oxygenators with respect to complement (C3a), granulocyte (lactoferrin) and monocyte (interleukin-6, IL-6) activation. Thirty patients undergoing cardiac surgery were randomly assigned to undergo cardiopulmonary bypass (CPB) with one of the following oxygenators: a polypropylene hollow-fibre membrane (group 1; 2.2 m2), a polypropylene flat-sheet membrane (group 2; 3.1 m2) or a silicone envelope membrane (group 3, 3.5 m2). In all patients, a significant increase in C3a in plasma occurred during CPB with peak levels after the administration of protamine sulphate. In blood samples taken before aortic crossclamp release, at the end of CPB, and 20 min after protamine administration C3a was significantly lower in group 1 than in the other two groups. Lactoferrin increased significantly during CPB in all patients without a significant difference between the groups. IL-6 did not increase during CPB, but raised significantly after 4 h in the intensive care unit in all groups. Moreover, IL-6 was significant lower in group 1 than group 3. The data suggest that the polypropylene hollow-fibre membrane oxygenator, i.e. the oxygenator with the smallest surface area, is more biocompatible than the other types, probably because of a smaller contact surface area. PMID- 9413850 TI - Potential problems with simplified selective cerebral perfusion--experimental investigations and clinical improvements. AB - Currently the most used perfusion techniques during aortic arch surgery to prevent cerebral damage include hypothermic circulatory arrest, retrograde cerebral perfusion and selective cerebral perfusion (SCP). The application of simplified SCP, which does not require deep hypothermia, has become an alternative procedure for brain protection. Including the physiological principle of autoregulated cerebral blood flow, cerebral perfusion flow is not predetermined, but differentiated from the different cannula sizes for the lower and upper body perfusion. In a mock circulation loop, we could show that resistance changes in the two compartments led to flow shifts between the systemic and brachiocephalic regions. In addition to mechanical factors cerebral perfusion is determined from physiological changes. In practice, these shifts can be initiated with disrupted autoregulation due to ph-stat management or dramatic pressure changes. To prevent mismatched cerebral perfusion extended perioperative monitoring was included in our clinical setting. With bilateral somatosensory evoked potentials, a computer aided topographical electroencephalometry system, transcranial doppler-sonography and jugular venous bulb saturation, we could provide a sufficient bihemispheric perfusion. Between 1990 and 1995 we operated on 21 patients using SCP. Intraoperatively no signs of cerebral ischaemia due to inadequate perfusion could be observed. Only temporary neurological changes were found postoperatively. In summary, the simplified SCP, despite its physiological basis, is intricately involved in control and influence. We think that the application of SCP is safe if extended neurophysiological monitoring is included in the clinical setting. PMID- 9413851 TI - Does heparin coating improve biocompatibility? A study on complement, blood cells and postoperative morbidity during cardiac surgery. AB - To evaluate whether the effect of heparin coating the extracorporeal circuit resulted in differences in patient outcome and haemostatic alteration, 24 patients undergoing elective, isolated coronary artery bypass were randomized prospectively to cardiopulmonary bypass (CPB) with heparin-coated circuits (group H, n = 12) or uncoated circuits (group C, n = 12). The technique of CPB, heparinization and its reversal were the same in both groups. We studied complement status (C3d, C3, C3d/C3, C4 and C-function), white blood cell counts with differentiation and the postoperative morbidity. The results confirmed that CPB activates complement and increases neutrophils in both the H and C groups. A significantly lower level of leucocytosis was seen in group H compared to the C group (p < 0.05). The complement function via the classical pathway (C-function), expressed as a percentage of the function of a reference serum pool (the values of normal sera were 75-125%), was significantly reduced in both heparin-coated and uncoated circuits (p < 0.05). There was no significant intergroup difference regarding C3, C3d/C3, C4 and C-function during the study period. A lower frequency of postoperative morbidity was present in the H group. We conclude that heparin-coated surfaces elicit less leucocytosis and decrease postoperative morbidity in patients undergoing cardiac surgery but do not cause a significant difference regarding activation of the complement system as reported by many other investigators. PMID- 9413852 TI - Alterations of the cytokine network in patients undergoing cardiopulmonary bypass. AB - Cardiovascular surgery using extracorporeal circulation causes a systemic inflammatory response which often results in severe organ dysfunction and increased postoperative mortality. Advances in knowledge about the interactions of cytokines involved in the response to cardiopulmonary bypass (CPB) may improve the outcome of patients undergoing cardiac surgery. The purpose of our study was to investigate the fluctuations in cytokine production, during and after CPB. In 24 patients undergoing elective coronary artery bypass grafting, plasma levels of interleukins IL-2, IL-6, IL-10 and IL-12, soluble IL-2-receptor (sIL-2R), and transforming growth factor-beta (TGF-beta) were measured at eight time points before, during and after CPB, using a standardized enzyme-linked immunosorbant assay technique. There was a significant increase in plasma levels of IL-10, IL-6 and TGF-beta after weaning off CPB. The IL-2 plasma levels decreased after the onset of CPB until 24 h postoperatively (p < 0.05). Concentrations of sIL-2R decreased 20 min after the start of CPB until the end of the operation (p < 0.05). In the postoperative course, sIL-2R levels increased, with peak values 48 h after the end of the surgical procedure. The IL-12 levels decreased after weaning off CPB (p < 0.05) until 6 h postoperatively. The results of our study demonstrate an intraoperative-predominant immunosuppression, followed by an early postoperative immunological activation, combined with a distinct acute phase response. PMID- 9413853 TI - Altered activation of the L-arginine nitric oxide pathway during and after cardiopulmonary bypass. AB - The serum concentrations of nitrogen oxides, the stable metabolites of nitric oxide, were measured in 61 children during and after cardiopulmonary bypass (CPB) for surgery of congenital heart disease. Overall, there was a small reduction in serum nitrogen oxide concentrations during CPB, from a median of 27.5 (interquartile range 16.6-55.7) to 26.4 (15.3-40.6) mumol/l, followed by an increase in the following 24 h to 33.1 (21.3-46.7) mumol/l. The largest postoperative increases in nitrogen oxides occurred in children who developed renal impairment, or were treated with nitrovasodilators. There was no relationship between changes in serum nitrogen oxides intraoperatively and early changes in pulmonary vascular resistance, and a weak positive relationship between changes in serum nitrogen oxides and early postoperative changes in cardiac index (r2 = 0.09, p = 0.04). We found no evidence for increased activation of the L-arginine nitric oxide pathway during CPB; and the reduction in nitric oxide metabolites that occurred during CPB were of doubtful significance to pulmonary or systemic haemodynamic changes in the postoperative period. PMID- 9413854 TI - Quantitative evaluation of flow patterns in perfusion cannulae by a new magnetic resonance imaging method. AB - The arterial cannula is a critical part of any extracorporeal circulation system due to the high flow rates which must pass a small cross-sectional area, resulting in high blood velocities. The aim of this study was to examine whether high-field magnetic resonance scanning is applicable for detailed mapping of velocity fields around the tip of such arterial cannulae in vitro. The investigated cannula was an angled, open-tip traditional design aortic cannula with an internal tip diameter of 5.5 mm. The velocity fields were measured at two different flow rates (2 and 4 l/min) at various positions in the lumen and outside the cannula using magnetic resonance imaging (MRI). All three components of the velocity vectors were measured. The study showed that MRI can provide a clear quantitative visualization of the velocity field around the tip of arterial cannulae at lower flow rates. At higher flow rates it can provide information about localization of regions with turbulent or disturbed flow. PMID- 9413855 TI - Inhibition of growth factor mitogenicity and growth of tumor cell xenografts by a sulfonated distamycin A derivative. AB - Interference with growth factor-receptor interactions may have particular relevance in efforts to intervene clinically in both autocrine and paracrine aspects of malignancy. Suramin is a synthetic anticancer agent that works, in part, by blocking the binding of growth factors to their receptors. While initial clinical trials have been encouraging, its use in clinical applications is associated with significant toxicities. Suradista is a novel sulfonated distamycin derivative that is also effective at complexing and inactivating growth factors and cytokines while remaining relatively nontoxic. The goal of this study was to compare the antineoplastic properties of suramin and Suradista. To achieve this, the effects of these compounds on growth factor induced mitogenesis in normal mouse fibroblasts and human umbilical vein endothelial cells were examined, as well as their ability to inhibit the growth of NIH/3T3 cells that had been transformed by the introduction of a fibroblast growth factor (FGF) 1 coding region (residues 1-154) fused to the signal peptide of the hst/KS3 gene (sp-hst/KS3:FGF1-154). In each case, Suradista was more effective than suramin in inhibiting mitogenesis in normal cells, as well as the growth of the transformed cells. Furthermore, Suradista was also shown to be as effective as suramin at inhibiting the growth of sp-hst/KS3:FGF1-154-transformed NIH/3T3 xenografts grown in athymic nude mice when given at only 50% the dosage used for suramin (50 mg/kg for Suradista versus 100 mg/kg for suramin). In summary, these results indicate that novel compounds acting like suramin may be developed as effective antineoplastic agents and may also prove to be of clinical benefit. PMID- 9413856 TI - In vitro binding of MX2 (KRN8602) and epirubicin to human plasma protein. AB - This study compares the human plasma protein binding characteristics of MX2 and epirubicin. The binding characteristics were determined by equilibrium dialysis at various concentrations of the drugs. The binding dissociation constant (Kd), binding capacity (Bmax) and partitioning constant (Kp) were obtained by Scatchard analysis of the free and bound drugs in the dialysis compartments. Our results have demonstrated that plasma protein binds epirubicin or MX2 in an unsaturable appearance over the concentration up to 150 mumol/l. At the same concentrations, plasma protein binds more epirubicin than MX2. The nature of the interaction may consist of two classes of specific binding, and a partitioning. The binding dissociation constants were 18 and 17.5 mumol/l for the higher binding class (Kd1) and 315.8 and 316.9 mumol/l for the lower binding class (Kd2), respectively, for epirubicin and MX2. The respective maximum binding capacities (Bmax) of plasma protein for epirubicin and MX2 were significantly different, 0.045 and 0.029 mumol/g protein for the higher binding class (Bmax1), and 0.39 and 0.29 mumol/g protein for the lower binding class (Bmax2). The partitioning constants (Kp) were 21.5 x 10(-5) and 20 x 10(-5) litres/g protein for epirubicin and MX2, respectively. The results suggest that plasma protein binds epirubicin or MX2 with a similar affinity, but has less binding sites for MX2. One contributing mechanism to the difference in activity noted between epirubicin and MX2 may be changes in free drug fractions. PMID- 9413857 TI - Nitric oxide released from Swiss 3T3 fibroblasts acts as a cytostatic agent for cultured mast cells. AB - Nitric oxide (NO) released from Swiss 3T3 fibroblasts inhibited the proliferation of cultured mast cells (CMCs) on the CMCs/Swiss 3T3 fibroblast co-culture system. Swiss 3T3 fibroblasts produced NO upon treatment with recombinant interferon gamma (rIFN-gamma). The production of NO was markedly increased by the co treatment of rIFN-gamma and recombinant tumor necrosis factor-alpha. This production was dependent on L-arginine and could be inhibited by the competitive substrate inhibitor, L-arginine analogue NG-monomethyl-L-arginine (NGMMA). In addition, the number of CMCs increased when treated with NGMMA in the co-culture system. These findings indicate that the elevated production of NO from Swiss 3T3 fibroblasts during the co-culture inhibited the proliferation of CMCs. PMID- 9413858 TI - Sensitization to the locomotor stimulant activity of cocaine is associated with increases in nitric oxide synthase activity in brain regions and spinal cord of mice. AB - Effects of multiple administrations of cocaine and subsequent cocaine withdrawal on the activity of nitric oxide synthase (NOS) in brain regions and spinal cord of male Swiss-Webster mice were determined. Chronic administration of cocaine resulted in the development of sensitization to its locomotor activity in the mouse. Chronic administration of cocaine was associated with increases in NOS activity in cerebral cortex, cerebellum, midbrain, hypothalamus, hippocampus, amygdala and spinal cord, but NOS activity was unaffected in pons/medulla and corpus striatum. Forty-eight hours after withdrawal from cocaine, NOS activity was increased only in the cerebral cortex. Recent studies have shown that cocaine induced sensitization to behavioral effects can be inhibited by NOS inhibitors. The present studies provide the first evidence that chronic treatment with cocaine alters NOS activity in brain regions and spinal cord, and are consistent with behavioral studies with cocaine and NOS inhibitors. PMID- 9413859 TI - Contribution of endogenous endothelin-1 to the maintenance of vascular tone: role of nitric oxide. AB - Studies were designed to compare the effect of the nitric oxide inhibitor, N omega-nitro-L-arginine (L-NNA), and the novel ETB receptor antagonist, RES-701-1, on changes in blood pressure and renal blood flow induced by exogenous endothelin receptor agonists and to determine the effect of L-NNA on basal hemodynamics in conscious, chronically instrumented rats. Infusion of low (nonpressor) doses of L NNA or RES-701-1 potentiated systemic and renal vasoconstriction induced by bolus injections of endothelin-1 or sarafotoxin 6c. Bolus intravenous injection or sustained infusion of L-NNA alone resulted in dose-dependent increases in blood pressure and decreases in renal blood flow, similar to our recently reported results with RES-701-1. Vasoconstriction induced by inhibition of nitric oxide was attenuated by SB 209670, a mixed ETA/B receptor antagonist, but not by BQ123, an ETA receptor antagonist; neither antagonist altered basal hemodynamics. Collectively, the results indicate that: (1) endothelin plays an important role in the control of basal vascular tone by mediating both vasodilation and vasoconstriction; (2) these effects are mediated by different ETB receptor subtypes in the rat, one located on the endothelium that mediates vasodilation via the nitric oxide pathway, the other located on the vascular smooth muscle that mediates contraction. PMID- 9413860 TI - Subcellular distribution of SERCA and calcium-activated ATPase in rabbit and human urinary bladder smooth muscle. AB - Previous studies have demonstrated that calcium storage and release from IP3 dependent sites in the sarcoplasmic reticulum play an important role in the contractile response of the rabbit urinary bladder to both field stimulation (mediated via neurotransmitter release) and bethanechol (direct muscarinic stimulation). In view of the importance of SERCA (see text) in urinary bladder smooth muscle function, we studied the distribution of SERCA by two methods; using Western blotting to quantitate the protein concentration and by enzyme analysis using thapsigargin to specifically inhibit SERCA. Rabbit and human samples of urinary bladder smooth muscle were homogenized and the homogenate separate into three particulate fractions by different centrifugation: the cell wall-nuclear, mitochondrial, and microsomal. The protein concentration of these three particulate fractions was determined and the SERCA protein level quantitated by Western blotting using SERCA-2 antibodies. The calcium ATPase activity was quantitated using standard enzymatic analysis and the thapsigargin sensitivity determined. The results demonstrated that (1) the concentration of SERCA was significantly greater in the microsomal fraction than in either of the other fractions for both rabbit and human bladder smooth muscle; (2) the enzymatic activities of both total calcium-activated ATPase and thapsigargin sensitive calcium ATPase were evenly divided among the three fractions, and (3) the enzymatic activity of both total calcium-activated ATPase and thapsigargin sensitive calcium ATPase of the rabbit exceeded that of the human. In conclusion, the distribution of SERCA and calcium ATPase of the rabbit bladder smooth muscle was similar to that in the human bladder smooth muscle, although activities in rabbit were significantly greater than those of human tissue. PMID- 9413861 TI - Functional electrical stimulation. AB - In the health-care professions, electrical stimulation is used for three purposes: to aid diagnosis; as a therapeutic tool; and to restore lost or damaged functions. Functional electrical stimulation (FES) and functional neurostimulation (FNS) are terms which are more or less interchangeable, and which encompass the third of these purposes. FES itself can also be conveniently divided into three classes, according to purpose: the restoration of sensor functions; the restoration of skeleto-motor functions; and the restoration of autonomic functions. Potentially, a fourth class would comprise devices restoring cognitive or psychological functions, but no such devices are clinically available as yet. The methods and devices which are currently available for providing FES are reviewed, as are the sorts of result and benefit that may be expected from them. The structure and emphasis of the review is on the clinical applications and the relevant anatomical and neurophysiological considerations and this approach is chosen for two main reasons. Firstly, the clinical, anatomical, and physiological considerations are independent of technological change and development, so they will not become quickly out of date. Secondly, the author is a clinician by profession, and an engineer only by inclination. The functional aims of FES methods will continue to develop as a result of experience gained following the introduction and use of successful devices, and these evolutionary improvements will come from within the FES programme; but the engineering embodiment of those devices may be revolutionized at any time by technological advances coming from elsewhere. PMID- 9413862 TI - A new technique for improved densitometric quantification of coronary artery stenoses in angiographic images. AB - In vitro studies have demonstrated that densitometric quantification of coronary artery stenoses is superior to geometric methods to assess non-circular lumens. However, in patients, several authors have reported significant discrepancies between area reduction percentages obtained densitometrically from two different imaging projections. Some of the factors causing the discrepancies can be reduced by simple precautions taken during image acquisition. Some others may be compensated for during analysis. Nevertheless, two factors remain problematic. The first is the inadequate spatial orientation of the vessel axes at the stenotic and reference cross sections with respect to the x-rays. The second is the difficulty in identifying the same vessel cross section in both planes at the time of analysis. We have designed a new densitometric technique that eliminates the error contributions of these two factors. The technique requires simultaneously acquired biplane coronary angiograms and biplane images of a translucent cube bearing steel markers acquired in exactly the same biplane geometry. Using the two projection matrices calculated from the images of the cube, the centerlines and the edges of the coronary arteries can be reconstructed in space from the biplane angiograms. The angles between the vessel axes and the x-ray beams can be determined and the densitometric cross sections can be corrected accordingly. Moreover, the 3D reconstruction allows the identification of the same cross section in the two planes for the determination of the area reduction percentages. Validation measurements were performed on a Perspex phantom and in patients, before and after angioplasty. In both types of measurement, the interplane discrepancies could be roughly halved. The densitometric technique presented can be incorporated into routine angiography and could become a strong alternative to the geometric approach that is presently dominating this field. PMID- 9413863 TI - Assessment of errors in static electrical impedance tomography with adjacent and trigonometric current patterns. AB - In electrical impedance tomography (EIT), difference imaging is often preferred over static imaging. This is because of the many unknowns in the forward modelling which make it difficult to obtain reliable absolute resistivity estimates. However, static imaging and absolute resistivity values are needed in some potential applications of EIT. In this paper we demonstrate by simulation the effects of different error components that are included in the reconstruction of static EIT images. All simulations are carried out in two dimensions with the so-called complete electrode model. Errors that are considered are the modelling error in the boundary shape of an object, errors in the electrode sizes and localizations and errors in the contact impedances under the electrodes. Results using both adjacent and trigonometric current patterns are given. PMID- 9413864 TI - The performance of an infra-red interactance instrument for assessing total body fat. AB - Infra-red interactance has been evaluated as a technique for measuring total body fat in comparison with a range of alternative methods. The alternative techniques employed were neutron activation analysis, tritiated water dilution, whole-body potassium-40 counting, skinfold anthropometry, bioelectrical impedance analysis and the body mass index. The study group consisted of 43 healthy adults (16 males and 27 females). For 11 women, measurements were obtained before and after 11 weeks on a very low-calorie diet, giving a total of 54 sets of data. Correlation coefficients between infra-red interactance and the other techniques varied between 0.58 (p < 0.0002) and 0.80 (p < 0.0001) for females, and between 0.64 (p < 0.009) and 0.94 (p < 0.0001) for males. The average fat for the study group was underestimated by 15% using infra-red interactance in comparison with the average fat obtained from the other techniques. It was also noted that the infra-red interactance instrument yielded a very narrow range of body fats in females in comparison with the other techniques. It is essential that these differences are reconciled before infra-red interactance takes a significant role in body composition analysis. PMID- 9413865 TI - Using automated analysis of the resting twelve-lead ECG to identify patients at risk of developing transient myocardial ischaemia--an application of an adaptive logic network. AB - The aim of this study was to introduce an adaptive logic network computing method for detecting patients who were likely to show transient ischaemic episodes during ambulatory Holter monitoring, using parameters from a previously recorded standard twelve-lead resting electrocardiogram (ECG). In the present study, the adaptive logic network computing method is compared with other commonly used classification methods, such as backpropagation network and discriminant analysis techniques. Of 1367 study subjects aged 65 and above, 733 were women and 634 were men. Ambulatory Holter recordings were made to detect episodic ischaemia in study patients. Those subjects showing ischaemic episodes were classified as 'ischaemic' patients, and the remaining subjects were 'non-ischaemic'. Accuracy was 67% using the adaptive logic network computing method, 56% using the backpropagation network computing method, and 65% using statistical discriminant analysis. We concluded that the adaptive logic network technique offers a slightly higher accuracy and shows several potential advantages for automated detection of ischaemia in resting electrocardiograms. PMID- 9413866 TI - Non-invasive assessment of cardiac output during exercise in chronic obstructive pulmonary disease: comparison of the CO2-rebreathing method and electrical impedance cardiography. AB - In exercise testing of patients with chronic obstructive pulmonary disease (COPD), non-invasive assessment of stroke volume (SV) and cardiac output (CO) would be valuable. Electrical impedance cardiography (EIC) has proved to be a valid and reliable instrument in healthy subjects. In this study it is investigated whether this also applies to patients with COPD. In 19 COPD patients simultaneous SV measurements were performed during steady-state exercise using the CO2-rebreathing method and EIC (using a fixed blood resistivity value (rho = 135 or 150 omega cm: EIC-135 and EIC-150) or a haematocrit based rho (EIC-ht)). Although close correlations were found (overall correlation between CO2 rebreathing and EIC-ht: R = 0.92 for CO, R = 0.79 for SV), SV and CO measured by means of EIC were significantly higher at low-intensity exercise and lower at high-intensity exercise. The mean differences between the CO2-rebreathing method and EIC-ht were 0.55 ml for SV and 0.01 l min-1 for CO (overall exercise data). The limits of agreement (2SD of the mean difference) were 24.7 ml for SV and 2.56 l min-1 for CO. These figures are comparable to what is found when healthy subjects are studied. CO was closely correlated to oxygen uptake using the CO2 rebreathing as well as the EIC method; the slope of the regression line was closer to what has been reported in the literature with EIC. Results were better with the EIC-ht than with the EIC-135 and EIC-150 methods. It is concluded that EIC is a reliable and valid method for measurements of SV and CO in COPD during exercise. PMID- 9413867 TI - Heart rate variability spectral indices for haemodynamic classification of haemodialysis patients. AB - The usefulness of spectral indices extracted from the heart rate variability (HRV) in discriminating between hypotension-prone and hypotension-resistant haemodialysis patients was investigated. In 30 patients, classified as hypotension resistant (stable group) or hypotension prone (unstable group), beat to-beat heart period was measured during haemodialysis sessions terminated without collapses. HRV was analysed in the frequency domain combining classic autoregressive spectral estimation with two eigen decomposition-based techniques: the reduced rank approximation (RRA) of the autocorrelation matrix and the Pisarenko harmonic decomposition (PHD). Five spectral indices were obtained: the ratio between the powers in the LF and HF bands (LF/HF), the same ratio calculated after application of RRA (LF/HFRRA), the frequency of the main oscillatory component of HRV estimated through PHD with a decomposition order equal to 1 (F1) and equal to 2 (F2) and the difference between the frequencies of the two oscillatory components resolved in the latter cas (Fd). The performances of these indices in discriminating between the two groups of patients were evaluated estimating the misclassification probability (Pm) of a Bayesian quadratic classifier. The HRV spectral pattern was markedly different: in the stable patients power was mainly in the low-frequency band, whereas in the unstable group it was mainly in the high-frequency band. The frequency of the main oscillatory component was significantly greater in the unstable group than in the stable one. Spectral indices displayed good discrimination power, increasing with the length of the dialysis interval. Best performances were achieved by LF/HFRRA both over short dialysis periods (Pm approximately 12% over 20 min intervals) and over longer periods (Pm = 3.3% over 160 min); similar results were obtained with Fd over short periods and LF/HF over long periods. Spectral HRV indices demonstrate, therefore, a diagnostic value in discriminating between hypotension-resistant and hypotension-prone patients. PMID- 9413868 TI - A non-invasive system to evaluate diaphragmatic strength in ventilated patients. AB - In this paper we describe a system that allows an indicator of diaphragmatic strength to be determined non-invasively in mechanically ventilated patients. The system is comprised of an occlusion device that can be incorporated into the ventilator tubing and an occlusion control unit to operate the occlusion. The system electronically coordinates the timing of airway occlusion at the mouthpiece, application of a magnetic phrenic nerve stimulus and rapid removal of the occlusion once the measurement has been made. The system therefore permits measurement of the change in airway pressure produced by a stimulated diaphragm contraction, without disconnection from the ventilator. Other important respiratory measurements such as twitch mouth pressure, simulated cough and P0.1 can also be performed on non-ventilated patients in a repeatable and systematic manner. PMID- 9413869 TI - An investigation of lower oesophageal redox potentials in gastro-oesophageal reflux patients and healthy volunteers. AB - Oesophageal electrical properties are thought to be important in the development of gastro-esophageal reflux. This study simultaneously monitored the intraoesophageal pH and redox potentials in 18 patients with gastro-oesophageal reflux symptoms and 15 asymptomatic controls, for a 24 h period. The pH and redox electrodes were positioned 5 cm proximal to the lower oesophageal sphincter, the position of which had been determined by manometry. Since significantly different behaviour was observed during the day and night, the data were divided into periods of waking and sleeping. Data were analysed for acid reflux (pH < 4) and transients in the redox potential-time curve. Both patients and normal subjects showed negative redox transients which were more frequent and pronounced at night than during the day, and which were uncorrelated with acid reflux. The only parameter which was significantly different between normal and refluxing groups was the amount of nocturnal redox activity, which was lower in refluxing subjects than in normals. Some possible hypotheses for these observations, and the origin of the redox species, are discussed. PMID- 9413870 TI - Value of simulated body surface potential maps as templates in localizing sites of ectopic activation for radiofrequency ablation. AB - Body surface potential maps recorded during catheter pace mapping can facilitate the localization of the site of origin of ventricular tachycardia. In this study, we investigated the value of a realistic computer model of the human ventricular myocardium in generating body surface potential maps as templates for identifying sites of ectopic activation. Our model features an anatomically accurate geometry and an anisotropy due to transmural fibre rotation, that were reconstructed with a spatial resolution of 0.5 mm. It simulates the electrotonic interactions of cardiac cells by solving a nonlinear parabolic partial differential equation, but it behaves as a cellular automaton when the transmembrane potential exceeds the threshold value. We successfully validated our model by comparing the simulated activation sequences--described by isochronal maps, epicardial potential maps and body surface potential maps--with the measured sequences of epicardial and body surface maps reported in the literature. By systematically pacing the left ventricular and right ventricular endocardial surfaces in our ventricular model, we generated a database of 155 QRS-integral maps, which provides a high resolution reference frame for localizing distinct endocardial pacing sites. This database promises to be a useful tool in improving the performance of catheter pace mapping used in combination with body surface potential mapping. Overall, the results demonstrate that our computer model of the human ventricular myocardium is well suited for complementing a database of QRS-integral maps obtained during clinical pace mapping and can help enhance the efficacy of the ablative treatment of ventricular arrhythmias. PMID- 9413871 TI - In vitro estimation of foetal liver volume using ultrasound, x-ray computed tomography and magnetic resonance imaging. AB - Sixteen formalin-fixed foetal livers were scanned in vitro using a new system for estimating volume from a sequence of multiplanar 2D ultrasound images. Three different scan techniques were used (radial, parallel and slanted) and four volume estimation algorithms (ellipsoid, planimetry, tetrahedral and ray tracing). Actual liver volumes were measured by water displacement. Twelve of the sixteen livers also received x-ray computed tomography (CT) and magnetic resonance (MR) scans and the volumes were calculated using voxel counting and planimetry. The percentage accuracy (mean +/- SD) was 5.3 +/- 4.7%, -3.1 +/- 9.6% and -0.03 +/- 9.7% for ultrasound (radial scans, ray volumes), MR and CT (voxel counting) respectively. The new system may be useful for accurately estimating foetal liver volume in utero. PMID- 9413872 TI - Amine precursor therapy: manipulation of brain amine activity with precursor amino acid. AB - Amine precursor therapies for the treatment of neuropsychiatric disorders and their neurochemical background are reviewed historically. Furthermore, peculiar or artificial amino acids, which are metabolized into the brain amines or their derivatives and promised their clinical application of treatment for morbid conditions, are also introduced. PMID- 9413873 TI - Vitamin B12 treatment for delayed sleep phase syndrome: a multi-center double blind study. AB - The active form of vitamin B12 (methylcobalamin) has been reported to be effective on sleep-wake rhythm disorders. Previous studies, however, were performed under open trial, and the effect of vitamin B12 has not been properly evaluated. The aim of this double-blind study was to investigate the efficacy of methylcobalamin on delayed sleep phase syndrome (DSPS). Methylcobalamin (3 mg/day) or placebo was administered for 4 weeks. The subjects were 50 patients with DSPS aged 13-55 years (26.8 +/- 1.3), 27 of whom received the active drug while 23 received the placebo. No significant differences were observed between the 2 groups in subjective evaluations of mood or drowsiness during the daytime or in night sleep by sleep-log evaluation. These results indicate that 3 mg methylcobalamin administered over 4 weeks is not an effective treatment for DSPS. PMID- 9413874 TI - Reliability and validity of the Japanese version of the Zarit Caregiver Burden interview. AB - Despite a rapid increase in disabled elderly in Japan, the burden of the caregiver has not been properly assessed due to a lack of objective measurements. Our study was aimed at adapting and validating the Zarit Caregiver Burden Interview (ZBI) in Japan, which is one of the most widely used measurements for caregivers' burden in the United States. Sixty-six caregivers answered the self administered questionnaire, involving the Japanese version of the ZBI and questions regarding their caregiving situation. Our study demonstrated that the Japanese version of the ZBI had equally as high reliability and validity as the original version. The Japanese ZBI had a high test-retest reliability (r = 0.76) and internal consistency (Cronbach's alpha = 0.93). The total score of the ZBI was highly correlated with the caregivers' score of the Center for Epidemiologic Studies Depression Scale (CES-D) score (r = 0.50), as well as a single global rating of burden (r = 0.71). It was also shown that demographic distribution of the score of the Japanese version had a similar trend to that of the original version. Caregivers who looked after patients with behavioral disturbances were found to have a significantly higher ZBI score than those who looked after patients without behavioral disturbances, which is consistent with previous findings. It is concluded that the Japanese version of the ZBI can be used to measure feelings of burden of caregivers in the Japanese population and can be used for cross-cultural comparison. PMID- 9413875 TI - Longitudinal study of the mental health of caregivers caring for elderly patients with dementia: effect of institutional placement on mental health. AB - One hundred and three family caregivers of relatives with dementia were longitudinally surveyed to examine the course of caregivers' mental health after the relatives had been placed in full-time care facilities. Mental health was assessed twice, with a 6-month interval, using the Japanese version of the 60 item General Health Questionnaire. The degree of social dysfunction was significantly reduced within 6 months after placement, while the other indicators of mental health (e.g. anxiety-insomnia, depression, somatic symptom and psychiatric morbidity) were not reduced within this term. However, anxiety insomnia and psychiatric morbidity were significantly reduced more than 6 months after placement. Analyses by caregivers' lineal relations to the relatives indicated that only daughters-in-law showed a significant decrease in anxiety insomnia and a marginal reduction in psychiatric morbidity. This study suggests that caregivers' social dysfunction was more greatly reduced than anxiety insomnia and psychiatric morbidity within a relatively short term after placement, and that its effects on mental health might vary with the lineal relations. PMID- 9413876 TI - Neglect in elderly stroke patients: a comparison of five tests. AB - Neglect is a disabling state in stroke patients. Five tests for visuo-spatial neglect, star cancellation, line crossing, line bisection, draw a clock and copy a cross, were compared in 57 elderly patients. Sensitivity and intercorrelations between the tests were determined. Patients with neglect were studied as regards their activities of daily living, motor activity and cognition. Left-hemisphere neglect in patients with right-hemisphere lesions was more severe than right sided neglect in patients with left-sided lesions in the star cancellation test. Sensitivity of the tests was moderate for star cancellation, line bisection and draw a clock, and low for line crossing and copy a cross. Significant correlations existed between the tests and cognitive and functional ability. Intercorrelations between the tests were moderate. Neglect patients showed a slower recovery after 6 and 12 months. Draw a clock and a cross displayed no sensitivity for neglect in patients with hemianopia, which may be due to the fact that these tests also evaluate constructional apraxia. The cancellation tests were influenced by hemianopia and there is evidence that hemianopia exacerbates neglect. This study shows that a battery of neglect tests is required to diagnose the neglect syndrome. PMID- 9413877 TI - Kidney transplantation and liaison psychiatry, part I: anxiety before, and the prevalence rate of psychiatric disorders before and after, transplantation. AB - We examined psychiatric problems before and after kidney transplantation in a sample of 36 patients with end-stage renal failure. The prevalence rate of psychiatric disorders was 11.1% (4 of 36 cases) before the transplantation and 36.1% (13 of 36 cases) within 2 months after the transplantation. Except for a patient with schizophrenic disorder, no patients were found to have a psychiatric disorder from 2 to 6 months after the transplantation. In this study, we also examined anxieties and/or conflicts related to the transplantation using the synthetic house-tree-person (HTP) drawing test, a measure of mood states by means of a non-verbal expression method. The upper part of the tree trunk was not drawn in 25% of this sample (5 of 20 cases). In the HTP drawing tests immediately after the transplantation, however, trees missing the upper part of trunk were not drawn. Based on these findings, we discussed psychiatric problems in kidney transplantation. PMID- 9413878 TI - Kidney transplantation and liaison psychiatry, part II: A case of dissociative identity disorder. AB - The authors examined the case of an adolescent patient with dissociative identity disorder secondary to psychological shock of a transplant rejection response. Psychiatric symptoms consisted of three components: visual hallucinations and delusions as a psychological defense against the anxiety of a transplant rejection; appearance of three personalities including proper self, the dead child (donor), and a prophet with strong predicting power; and a twilight state. These psychiatric symptoms may have been related to two psychological factors: immature personality characteristics formed during hemodialysis, and post traumatic stress caused by a chronic rejection reaction from the patient's first transplant. PMID- 9413879 TI - Modifying psychotropic drug prescription patterns: a follow-up survey. AB - Cross-sectional surveys of prescription patterns of psychotropic drugs provide a quick, global estimation of the appropriateness of psychopharmacotherapy. Recurrent inadequacies in prescribing for psychiatric patients include polypharmacy, high doses of antipsychotics (APS), the use of multiple APS simultaneously, and the administration of these drugs in multiple divided doses. Faulty prescribing patterns are difficult to amend. Following a survey in a rehabilitation facility for chronic psychiatric patients, a systematic education program and other measures were introduced to improve prescribing habits. Two years later the survey was repeated. Significant improvements were detected in the following areas: reductions in the doses of APS and antiparkinsonian (AP) drugs, reductions in the number of patients placed on these drugs, and reductions in the percentage of patients receiving multiple antipsychotics simultaneously. Despite these reductions in doses of psychotropic agents, patients' rehabilitative potential was not compromised. PMID- 9413880 TI - MRI-based morphometric topographic parcellation of human neocortex in trichotillomania. AB - The purpose of the present study was to test specific hypotheses regarding volumetric changes of the neocortex between 10 female trichotillomania (TTM) subjects and 10 female normal controls. A standard three-dimensional (3-D) brain coordinate system was imposed over each newly acquired native magnetic resonance imaging (MRI) scan for positional normalization and 3-D shape/geometric localization analyses were based on the midpoints of anterior and posterior commissures, and the longitudinal fissure. The brain segmentation method, using well-characterized semiautomated intensity and differential contour algorithms by signal intensity-frequency histograms, was used blind to segment the principal gray and white matter structures. The segmented neocortical ribbon was subdivided into 48 regions (i.e. parcellation units) per hemisphere via a new method of morphometric topographic parcellation. There were no significant volumetric changes of the precentral gyrus, postcentral gyrus, supplementary motor cortex or opercular cortex in TTM patients compared with control subjects. A broader analysis as a hypothesis-generating post-hoc effort showed that TTM subjects exhibited significantly reduced left inferior frontal gyrus volume of 27% (t = 2.21, d.f. = 18, P = 0.04) and enlarged right cuneal cortex volume of 40% (t = 2.30, d.f. = 18, P = 0.03) compared to normal controls. This is the first report of a structural neocortex abnormality in TTM. Results are discussed in terms of the behavioral specialization of these two brain neocortical regions and the complex interractions between visual and sensorimotor cortices. The results also showed the feasibility of the MRI-based morphometric topographic parcellation for investigation of the human neocortex in neuroscience research. PMID- 9413881 TI - Deep venous thrombosis of the leg due to psychiatric stupor. AB - We report the cases of two patients with psychiatric stupor who developed venous thrombosis. A 29-year-old schizophrenic woman had been hospitalized in psychiatric institutions three times because of stupor associated with auditory hallucinations and thought blocking. These symptoms recurred and she was admitted to our hospital with deep venous thrombosis of her left leg. The other patient was a 67-year-old woman with depression. She had also suffered from insomnia. Following admission to our hospital, she developed a depressive stupor complicated by deep venous thrombosis of her left leg. Both cases were treated with sodium heparin and urokinase, and completely resolved. It is well known that dehydration, infection and decubitus ulcers are important physical complications of psychiatric stupor, but there have been few reports of deep venous thrombosis as a physical complication of stupor. PMID- 9413882 TI - A study of verbal and spatial information processing using event-related potentials and positron emission tomography. AB - The activated cerebral regions and the timing of information processing in the hemispheres was investigated using event-related potentials (ERP) and regional cerebral blood flow (rCBF) as the neurophysiological indicators. Seven men and one woman (age 19-27 years) were asked to categorize two-syllable Japanese nouns (verbal condition) and to judge the difference between pairs of rectangles (spatial condition), both tests presented on a monochrome display. In the electroencephalogram (EEG) session, EEG were recorded from 16 electrode sites, with linked earlobe electrodes as reference. In the positron emission tomography (PET) session, rCBF were measured by the 15O-labeled H2O bolus injection method. Regions of interest were the frontal, temporal, parietal, occipital and central lobes, and the entire cerebral hemispheres. When the subtracted voltages of the ERP in homologous scalp sites were compared for the verbal and spatial conditions, the significant differences were at F7.F8 and T5.T6 (the 10-20 system). The latencies of the differences at T5.T6 were around 200, 250 and 320 ms. A significant difference in rCBF between the verbal and spatial conditions was found only in the temporal region. It was concluded that early processing of information, that is, registration and simple recognition, may be performed mainly in the left temporal lobe for verbal information and in the right for spatial information. PMID- 9413884 TI - A case of somatic delusional disorder that responded to treatment with risperidone. PMID- 9413883 TI - An association study between a transcriptional polymorphism in the serotonin transporter gene and panic disorder in a Japanese population. AB - A polymorphism in the 5' region of the serotonin transporter gene modulates its transcription efficiency. Its short allele has been reported to be associated with neurotic traits. The serotonin transporter is the action site of selective serotonin re-uptake inhibitors, widely used in the treatment of panic disorder. We examined an association between the polymorphism and panic disorder in a case control study consisting of 66 Japanese patients and 150 controls. The short allele was significantly more frequent in the Japanese than in Caucasians. The patients and the controls had similar allele frequencies, indicating no association between the polymorphism and panic disorder in most Japanese patients. PMID- 9413885 TI - Spinal instability: fact or fiction. PMID- 9413886 TI - Treatment of calcific tendinitis and adhesive capsulitis of the shoulder. PMID- 9413887 TI - Glucocorticoid-induced osteoporosis: novel approaches. PMID- 9413888 TI - New autoantibodies in rheumatoid arthritis. PMID- 9413889 TI - Vertebroplasty: current data and future potential. PMID- 9413890 TI - Significance of leukotriene-A4 hydrolase in the pathogenesis of psoriasis. AB - The 5-lipoxygenase (5-LO) product of arachidonic acid, leukotriene (LT-)B4, is considered to play a significant role in the pathogenesis of psoriasis. In vitro LTB4 is a potent chemoattractant for leukocytes, and it increases DNA synthesis in human cultured keratinocytes. Intradermal injection of LTB4 into human skin in vivo results in a wheal and flare reaction, and topical application produces intraepidermal microabscesses and induces hyperproliferation. Furthermore, LTB4 has been determined in biologically active amounts in psoriatic skin lesions. Despite the importance of LTB4 in psoriasis, the capacity of the human epidermis to synthesize LTB4 has remained controversial. Recently, a very limited 5-LO activity was reported in human epidermis. Thus, it was shown that human epidermis can contribute significantly to LT formation by transcellular LT synthesis. By this mechanism, LTA4 released from activated leukocytes is further transformed into LTB4 in the keratinocytes by the LTA4 hydrolase. Transcellular metabolism may be of importance in psoriasis where neutrophils migrate into the epidermis, because in human neutrophils the LTA4 hydrolase has been shown as the rate limiting step in LTB4 formation. The LTA4 hydrolase was localized in the epidermis by activity determination, by inhibition of enzyme activity with known LTA4 hydrolase inhibitors, by Western blotting and by immunohistochemical staining. Moreover the enzyme was purified and further characterized from human cultured keratinocytes and human epidermis. Because of these recent results it is concluded that LTB4 is of significance in the pathogenesis of psoriasis, and it is suggested that future work should focus on developing potent LTA4 hydrolase inhibitors for treatment of psoriasis. PMID- 9413891 TI - Effect of percutaneous absorption of fluocinolone acetonide on the activity of superoxide dismutase and total antioxidant status in patients with psoriasis. AB - This study defines a modification of antioxidant systems by percutaneous absorption of fluocinolone acetonide. Total antioxidant status (TAS) provides an overall indication of antioxidant status. Superoxide dismutase (SOD), a primary antioxidant, accelerates the dismutation of the toxic superoxide radical produced during the oxidative energy processes into the less harmful molecules, hydrogen peroxide and molecular oxygen. We monitored the level of SOD and TAS in 7 males with psoriasis and 6 control subjects before and after a single application of fluocinolone acetonide 0.025% ointment to 90% of the body. The results showed that the plasma level of TAS was significantly increased (p < 0.02) at 24 h posttreatment. The erythrocytic level of SOD was significantly decreased (p < 0.01) only at 12 h after glucocorticosteroid application. The level of TAS and SOD in patients with psoriasis was also significantly increased (p < 0.01 for both situations) as compared to healthy controls. Our study suggests that fluocinolone acetonide as a therapeutic agent may play a role in the oxidative stress in skin diseases. PMID- 9413892 TI - Experimentally induced pruritus and cutaneous reactions with topical antihistamine and local analgesics in atopic eczema. AB - We investigated the antipruritic effect of a 15-min application of dimethindene maleate (Fenistil gel) and other local analgesics (Optiderm, EMLA, Xylocain ointment 5%) on subsequent focal histamine stimulus (20 mC) given by iontophoresis in 12 patients suffering from acute atopic eczema (AE). The results were compared to histamine after pretreatment with the respective placebo and to non-pretreated skin. Wheal and flare areas were planimetrically evaluated. Itch or pain ratings were performed over a 24-min period using a rating scale. The examination also comprised alloknesis, i.e. induction of a perifocal itch sensation by a non-itching mechanical stimulus. None of the antihistaminic and anaesthetic agents reduced the itch intensity significantly. Three of the AE patients had a total lack of alloknesis. We conclude that these substances, when applied for 15 min, are not sufficiently effective in atopic skin suppressing histamine-induced reactions under experimental conditions. The diminished elicitation of alloknesis in these patients may be a result of central nervous system alteration. PMID- 9413893 TI - Inhibition of skin protein kinase C by psychotropic drugs. AB - Lipid-soluble psychotropics are often used to treat skin diseases with psychosomatic indications. Although these drugs are known to exert their effects through the central nervous system, relatively little is known about their mechanism of action in skin. In this communication, several lipid-soluble psychotropic drugs have been examined for their ability to inhibit protein kinase C (PKC)-catalyzed phosphorylation of exogenous substrates and endogenous skin proteins. Phosphorylation of three discrete skin protein substrates at 64, 42 and 28 kDa and a group crowded together at 15-18 kDa was prevented by the antidepressants/antipsychotics. Inhibition was more pronounced in a phospholipid (PL) dependent system, but both drug-PL and drug-PKC interactions seem to be important in the mechanism of action of these drugs. In addition to the tricyclic nucleus, the propanamine side chain or its N-methyl form may influence the interaction of these drugs with PKC and its substrate(s). Chlorpromazine, imipramine, fluoxetine, doxepin, amitriptyline and hydroxyzine used in the practice of dermatology may exert their therapeutic effects by modulating skin PKC activity. PMID- 9413894 TI - Effects of flavonoids of Ginkgo biloba on proliferation of human skin fibroblast. AB - Ginkgo biloba studies have focused on the anti-inflammatory effects of the major components, ginkgolide and bilobalide, whereas little is known about their effect on fibroblasts. This study demonstrated the enhancing effects of Ginkgo L. extracts, especially the flavonoid fractions: quercetin, kaempferol, sciadopitysin, ginkgetin, isoginkgetin, on the proliferation of normal human skin fibroblast in vitro measured by MTT (3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyl tetrazolium bromide) assay and direct hemocytometer cell count. Furthermore, increased production of collagen and extracellular fibronectin were documented by radioisotope (2,3-3H-proline) incorporated collagen assay, procollagen type I C peptide assay and by immunoturbidimetric assay. These proliferative effects suggest another useful pharmacologic application of Ginkgo L. extracts in addition to their well-known anti-inflammatory effect. PMID- 9413895 TI - Biphasic effects of minoxidil on the proliferation and differentiation of normal human keratinocytes. AB - Minoxidil is the most used drug with proved effects in the treatment of androgenetic alopecia (AGA), but little is known about its pharmacological activity and target cells in hair follicles. As AGA is characterized by follicle atrophy, accelerated hair cycles and hair fiber thinning, we postulated that keratinocyte proliferation/differentiation is affected and we tested Minoxidil's effects on those parameters. Normal human keratinocytes (NHK) of follicular or epidermal origin were cultured in the presence of Minoxidil (0, 0.1, 1, 10, 100, 1,000 microM) during 5-8 days in various media (high-/low-calcium content, with or without serum). Proliferation was assessed by mitochondrial dehydrogenase activity (XTT), BrdU incorporation, lysosome numeration (neutral red incorporation) and total protein dosage. Drug-induced cytotoxicity was measured by lactate dehydrogenase release in culture supernatant, and pro-differentiating effects were evaluated by relative involucrin expression (ELISA dosage). On this basis, we showed that Minoxidil had biphasic effects on the proliferation and differentiation of NHK: Minoxidil stimulated NHK proliferation at micromolar doses, while antiproliferative, pro-differentiative and partially cytotoxic effects were observed with millimolar concentrations. We can hypothesize that Minoxidil hypertrichotic activity in vivo is possibly mediated by the maintenance of proliferative potential in follicular keratinocytes precociously committed to differentiation. PMID- 9413896 TI - Influence of indoles (melatonin, serotonin and tryptophan) on the porphyrin metabolism in vitro. AB - We examined the influence of melatonin, serotonin and tryptophan on the basal and delta-aminolevulinic acid (ALA)-induced porphyrin content in HaCaT, SKMel-23 and HepG2 cells. ALA-preincubated and ALA-free cells were fed with medium containing 1 mM melatonin, serotonin or tryptophan. After 24 h the porphyrin content in the cells and in the culture medium was measured. In the three cell lines the inbucation with 1 mM ALA over 24 h increased the porphyrin concentration in all cell lines in different degrees: HepG2 > SKMel-23 > HaCaT cells. In HepG2 cells, neither melatonin, serotonin nor tryptophan influenced ALA-induced porphyrin concentrations significantly, but all three indoles depressed the porphyrin levels in SKMel-23 and HaCaT cells. The indoles may decrease the ALA uptake in HaCat or SKMel-23 cells. Another mechanism could be the inhibition of enzymes converting ALA into porphyrins. PMID- 9413898 TI - From ancient roots to modern times. PMID- 9413897 TI - 75 years. PMID- 9413899 TI - Solid polymers: a challenge for NMR. AB - The sheer structural and motional complexity of polymers presents a formidable challenge to the power and versatility of NMR. This challenge is well met through exploitation of the plethora of experimental devices which have been developed over the past 50 years, among which the discovery of Magic Angle Spinning by Raymond Andrew features prominently. This paper presents a brief review of the subject in terms of a number of examples which illustrate the rich and detailed information which NMR provides. PMID- 9413900 TI - The rise of human in vivo NMR spectroscopy. AB - NMR spectroscopy and NMR imaging with magnetic field gradients make strange bedfellows, the requirements for one seemingly ruling out the other for human applications. Nevertheless, their stories are intertwined; the advent of high field imaging systems arose because of the desire for human spectroscopy. Localized spectroscopy is possible because of NMR imaging. Both have links to physics at Nottingham, at least in the personalized account that follows. Today, virtually all NMR spectroscopy experiments can be conceived with a localized in vivo spectroscopy counterpart. PMID- 9413901 TI - NMR of a rotating sample: the idea and consequences of a fruitful invention. PMID- 9413903 TI - Exchange interaction between nuclear spins. AB - We describe the developments in the determination of exchange interaction between nuclear moments in a metal. We discuss two techniques, the isotopic interference and higher harmonics of Larmor precession, both techniques determine the sign as well as the magnitude of the exchange constant. In addition, we briefly discuss the possibility of experiments at negative temperatures providing additional information about the space dependence of the exchange interaction. PMID- 9413902 TI - NMR and relative quantum theory. AB - Assumptions on which NMR dynamical studies have been based for nearly 50 years are consistent with a relative quantum theory based on Minkowski spacetime. A symmetry condition required by the existence of reference frames takes over the usual role of particle permutation symmetry, leaving a quantum theory of relative motion which only involves observables. PMID- 9413904 TI - High field NMR magnets. AB - Impressive progress has been made during past decades in many research fields which apply NMR, pushing the proton resonance frequency from 30 MHz (0.7 T) to about 800 MHz (19 T). In this overview we analyze different technologies for generation of the required fields and their potential for future developments toward even higher resonance frequencies of up to 2 GHz. PMID- 9413905 TI - 1 GHz NMR spectroscopy: innovation in magnet technology. AB - The present period is one of rapid development and major extension of the technology for high resolution and solid state NMR spectrometer magnets. Programs which have already been initiated have as their objective proton frequencies of 1 GHz and greater, eventually requiring HTS superconductors. These magnets will contain inner coils containing HTS conductors, surrounded by a set of relatively large coils fabricated with metallic superconductors. These coils represent a major extension of the adiabatically stable magnet technology that has evolved to address the performance issues posed by this type of magnet. The developments which are desirable for these large magnets are identified to include tough epoxy, interface to and thermal performance of external reinforcement, and high strength-high current density metallic superconductor. PMID- 9413906 TI - High resolution and high fields in biological solid state NMR. AB - 2H solid state NMR spectra of a polypeptide in an oriented membrane environment is demonstrated to have an orientational resolution of 0.3 degree. Such data results in high resolution structural constraints. Similar spectra are demonstrated at 23.2 T using a resistive magnet at the National High Magnetic Field Laboratory. PMID- 9413907 TI - Interdoublet transitions in S = 5/2 protein systems. AB - Beginning with known parameters that characterize the EMR spectra of several proteins containing high-spin ferric iron, the information content of the spectra has been examined by simulations that cover a range of magnetic fields and frequencies. Transitions between levels that are not Kramers doublet levels are particularly interesting when the applied frequency is approximately two to three times the value of the zero-field splitting parameter, D. In these cases, transitions at very low magnetic fields correspond to portions of interdoublet transitions that are well separated from all other transitions. The magnetic field is aligned at angles between the molecular principal axes for the portion of the molecules giving rise to the low-field interdoublet transitions. This provides an opportunity for unique angle-selection experiments. PMID- 9413908 TI - Spinning crystals leads to significant enhancement in 13C spectral resolution in MAS experiments on organic compounds: a new aid in studying phase transitions. AB - While Andrew [E.R. Andrew, Arch. Sci. (Geneva), 12 (1959) 103; E.R. Andrew, A. Bradbury and R.G. Eades, Nature, 183 (1959) 1802] utilized single crystals in his pioneering magic angle spinning (MAS) experiments, current studies use powders. Our recent MAS measurements on some organic compounds show that single crystals yield significantly narrower peaks than powders, especially for the 13C atoms that are not protonated. In our 13C CP (cross polarization)/MAS measurements on squaric acid (H2C4O4), for example, single crystals yielded a four-fold reduction in line widths as compared to its powder. This additional gain in resolution enabled us to differentiate between the order-disorder versus displasive contributions to the mechanism of the paraelectric-antiferroelectric phase transition of squaric acid at 373 K, which had not been possible with other techniques. Moreover, this gain in resolution appears to increase in proportion to the applied Zeeman field, while retaining the benefit of higher dispersion. PMID- 9413909 TI - A 5-year evaluation of ceramic inlays (CEREC). AB - The intention of this study was to evaluate the clinical function of porcelain inlays made with the Cerec technique after 5 years. 115 inlays were produced in 46 patients and of these were 51 inlays evaluated. Epoxy models were made from impressions and the gap width and depth was measured in a microscope. SEM pictures were also produced to illustrate the ditching. The measured marginal defects showed a mean width and depth of (SD) 373 (147) and 111 (67) microns respectively. No significant differences of the ditchings could be detected between molars and premolars. Only 3 inlays were found to be fractured of all inlays produced. PMID- 9413910 TI - Structural alterations in the TMJ disk in patients surgically treated for internal derangements (ADD). AB - Twenty-eight patients with temporomandibular dysfunction problem have been subjected to diskectomy unilaterally. The diagnosis was in 23 patients ADD (anterior disk displacement) without reduction and in another five ADD with reduction. The disk was removed in total according to a routine procedure. Surgical observation of a firm connection between the inferior surface of the disk and the condylar head was noted in eighteen patients. The disk tissue was orientated on a cork sheet according to its position in the fossa and subjected to histological examination. Histopathological analysis showed that the posterior attachment was a considerable part of the specimen in 12 out of 28 disks. Splitting of the disk was confined to the inferior surface. Chondrocytes were found close to the inferior surface. Surface irregularities, tags, were noticed on the inferior surface in thirteen cases. However, cellular signs of inflammation were found in the retrodiskal tissue in eight cases. PMID- 9413911 TI - Periodontal healing in horizontal and vertical defects following surgical or non surgical therapy. AB - The aim of the present study was to investigate any relationship between the level of oral hygiene and probing pocket depth reduction over time after periodontal treatment in sites with either vertical or horizontal destructions. The investigation was conducted as a retrospective study on a 3-year consecutive referral population of periodontitis-prone patients based on full-mouth oral radiographic examinations, probing pocket depth registrations and plaque scores. The analyses were performed on a final sample of 3064 sites in 107 patients with regression analysis after adjusting for dependence within the patient. Probing pocket depth was significantly less reduced over time in sites with vertical destructions compared to sites with horizontal destructions following non surgical treatment. Furthermore, the difference in probing pocket depth reduction between vertical and horizontal defects following non-surgical treatment increased over time in sites with plaque compared to sites without plaque, thus reflecting the importance of the patient's plaque control, especially in sites with vertical destructions. However, the difference in probing pocket depth reduction between vertical and horizontal defects did not increase over time for surgically treated teeth, a finding which probably can be attributed to a more thorough debridement of vertical defects during surgery and/or osteoplasty/osteoectomy limiting the surface area upon which a long junctional epithelium can form, which may facilitate recurrence of a periodontal pocket. PMID- 9413912 TI - Use of polymer materials in dental clinics, case study. AB - Dentistry uses a variety of different polymer materials. Dental polymer materials are based on methacrylate, its polymer, and polyelectrolytes. The setting of restorative materials and adhesives is initiated chemically by mixing two components or by light. In both cases, polymerisation is incomplete and monomers, not reacted, release. Studies have documented that monomers may cause a wide range of adverse health effects such as irritation to skin, eyes or mucous membranes, allergic dermatitis, asthma, parenthesise in the fingers, and disturbances from central nervous system such as; headache, pain in the extremities, nausea, loss of appetite, fatigue, sleep disturbances, irritability, loss of memory and changes in blood parameters. Dental personnel are occupationally exposed when handling the non reacted monomers. The use of gloves do not give enough protection as monomers, released from the material, easily penetrate all gloves used in dentistry. Face masks do not prevent inhalation of monomers. Ordinary glasses do not protect the eyes against vapor from monomers. The result from this study demonstrate the need for the development of ergonomic procedures and practices for safe handling of such materials in dental clinics. PMID- 9413913 TI - Analysis of the metal content of in vivo-fixed dental alloys by means of a simple office procedure. AB - The purpose of this study was to devise a simple technique to analyse the metal content of permanent dental prostheses without damaging the constructions. Metal debris was collected on grinding wheels for subsequent analysis by energy dispersive X-ray fluorescence (EDXRF). We discuss the importance of being able to qualitatively ascertain the composition of the alloys, as such information makes it possible to perform a detailed allergological investigation. Furthermore, we tentatively put our findings in a broader context with regard to the emerging knowledge of the relationship between the immune and central nervous systems as well as the known fact that certain metal ions can produce various cutaneous reactions. PMID- 9413914 TI - Comutagenesis-III. In vitro metabolism of 2-amino-3-methylpyridine: the effect of various potential inhibitors, activators and inducers. AB - 1. The effects of various potential inhibitors, activators and inducers on the metabolism of the comutagen 2-amino-3-methylpyridine (2A3MP) by rabbit hepatic microsomes and S9 supernatants have been studied. 2. The 1-N-oxidation of 2A3MP to 2-amino-3-methylpyridine-1-N-oxide (2A3M-PNO) was inhibited by 2,4-dichloro-6 phenylphenoxyethylamine (DPEA), 2-diethylaminoethyl-2,2-diphenylvalerate (SKF 525 A) and n-octylamine. 3. The C-oxidation products of 2A3MP, i.e. 2-amino-3 hydroxymethylpyridine (2A3HMP) and 2-amino-3-methyl-5-hydroxypyridine (2A3M5HP), were also inhibited by these compounds. 4. Pretreatment of animals with phenobarbitone (PB) resulted in an increase in the production of 2A3MPNO and 2A3HMP, whereas beta-naphthoflavone (BNF) pretreatment had a greater effect on the formation of 2A3M5HP. 5. Pretreatment with pyridine or pyrazine also had an appreciable effect on the formation of 2A3HMP. 6. It is suggested that different cytochrome P450 isozymes are responsible for the metabolic profile of 2A3MP. CYP2B was involved in the N-oxidation; 2E and/or 2B in the formation of 2A3HMP, and 3A and/or 1A in the formation of 2A3M5HP. PMID- 9413915 TI - Determination of the rate of aldicarb sulphoxidation in rat liver, kidney and lung microsomes. AB - 1. The rate of sulphoxidation of aldicarb (2-methyl-2-(methylthio) propanal O [(methylamino) carbonyl oxime], Temik) in rat hepatic, renal and pulmonary microsomes was determined by quantitating the levels of aldicarb sulphoxide and aldicarb sulphone produced during incubations. Under in vitro experimental conditions used in the present study, aldicarb sulphoxide was the only metabolite produced, and further metabolism of aldicarb sulphoxide to aldicarb sulphone was negligible. 2. The average maximal velocity (mumol/min/mg protein) for the sulphoxidation of aldicarb, based on measurements of product formation, in liver, kidney and lung microsomes was 5.41, 39.51 and 2.45 respectively. The corresponding values for the Michaelis constant (microM) were 184, 1050 and 188 respectively. 3. These results imply that under in vivo conditions (1) aldicarb sulphoxidation is not likely to be saturable even at lethal doses in the rat, and (2) aldicarb clearance in rat liver and kidney will be limited by the rate of blood flow and not metabolizing enzyme levels. PMID- 9413916 TI - Involvement of CYP2D1 in the metabolism of carteolol by male rat liver microsomes. AB - 1. The metabolism of carteolol, a beta-adrenoceptor blocking drug, was investigated in male Sprague-Dawley rat liver microsomes. 2. The formation of 8 hydroxycarteolol was the principal metabolic pathway of carteolol in vitro and followed Michaelis-Menten kinetics with a K(m) = 11.0 +/- 5.4 microM and a Vmax = 1.58 +/- 0.64 nmol/min/nmol P450 respectively (mean +/- SD, n = 5). Eadie-Hofstee plot analysis of carteolol 8-hydroxylase activity confirmed single-enzyme Michaelis-Menten kinetics. 3. The cytochrome P450 isoforms involved in 8 hydroxylation of carteolol were investigated using selective chemical inhibitors and polyclonal anti-P450 antibodies. Quinine (Ki = 0.06 microM) and quinidine (Ki = 2.0 microM), selective inhibitors of CYP2D1, competitively inhibited 8 hydroxycarteolol formation. Furthermore, only anti-human CYP2D6 antibody inhibited this reaction. 4. These results suggest that carteolol is metabolized to 8-hydroxycarteolol by CYP2D1. The K(m) of carteolol for CYP2D1 in male rat liver microsomes was much greater than those of propranolol or bunitrolol, indicating that carteolol has a lower affinity for CYP2D1 compared with these other beta-adrenoceptor blocking drugs. PMID- 9413917 TI - In vitro metabolic transformations of 2,4-dipyrrolidinylpyrimidine: a chemical probe for P450-mediated oxidation of tirilazad mesylate. AB - 1. We have determined that 2,4-dipyrrolidinylpyrimidine (2,4-DPP), used as a model for studies of the metabolism of therapeutic agents containing this moiety, undergoes three characteristic hydroxylations when incubated with male rat liver microsomes. Analysis of microsomal incubates of stable isotope labelled analogues of 2,4-DPP by particle beam-liquid chromatography-mass spectrometry (LC-PB-MS) has shown that the three metabolites are 4-(3-hydroxypyrrolidinyl)-2 (pyrrolidinyl)-pyrimidine (M1), 4-(2-hydroxypyrrolidinyl)-2-(pyrrolidinyl) pyrimidine (M2) and 2-(2-hydroxypyrrolidinyl)-4-(pyrrolidinyl)-pyrimidine (M3). 2. We determined that enzymes of the cytochrome P450 family are responsible for the in vitro hydroxylations of 2,4-DPP. 3. We observed that in microsomal incubations carried out in the presence of cyanide, a single cyanide adduct is formed implicating an iminium ion intermediate in the oxidation of the 2 pyrrolidine ring. 4. We also determined the intermolecular deuterium isotope effects for the formation of each of the three products. For M1, kH/kD = 14.55 +/ 0.54; for M2, kH/kD = 6.01 +/- 0.65; and for M3, kH/kD = 5.35 +/- 1.18. 5. We interpret these data as suggesting that M2 and M3 are formed by the same mechanism, probably including the formation of an iminium ion, and that M1 is formed by direct hydrogen abstraction. PMID- 9413918 TI - Stereoselective sulphate conjugation of fenoterol by human phenolsulphotransferases. AB - 1. The objective of this study was to determine (1) the molecular site(s) of sulphoconjugation of fenoterol; (2) the human phenolsulphotransferase (PST) isoform(s) involved; and (3) the stereochemistry of the enzymatic reaction. 2. Using the human Hep G2 cell line, hplc isolation and FAB/ms/ms, it was determined that fenoterol is sulphated both in the 4'-hydroxyphenyl position and in one of the 3',5'-dihydroxyphenyl positions. 3. Recombinant human M-PST preferentially sulphated the 4'-hydroxyphenyl position. In contrast, recombinant P-PST exclusively sulphated the 3',5'-hydroxyphenyl position. 4. The M-PST-catalysed sulphation of the 4'-hydroxyphenyl position was highly selective for the active RR-enantiomer, whereas the sulphation of the 3',5'-dihydroxyphenyl position was slightly selective for the opposite SS-enantiomer. 5. The P-PST-catalysed sulphation of the 3',5'-hydroxyphenyl position was selective for the inactive SS enantiomer. PMID- 9413919 TI - Species differences in the stereochemistry of the metabolism of isoprene in vitro. AB - 1. Comparative studies on the stereochemistry of the metabolism of isoprene in vitro have been carried out using liver microsomes from rats, mice, monkeys, dogs, rabbits and humans. Differences between strains and gender were also investigated. 2. In the production of the isoprene monoepoxides, microsomes from the livers of the male Sprague-Dawley or Wistar rat showed an approximately 2:1 preference for the formation of (S)-2-(1-methylethenyl)oxirane compared with the (R)-enantiomer. No enantioselectivity was observed for mouse or rabbit. In contrast, liver microsomes from dog, monkey or male human preferentially formed (R)-2-(1-methylethenyl)oxirane. There was no enantioselectivity observed with microsomes from female human liver. 3. The significant differences between species in the in vitro metabolism of isoprene indicate that stereochemical and mechanistic data should be taken into account when evaluating the results of animal studies designed to assess the carcinogenic risks to humans that may be associated with exposure to isoprene. PMID- 9413920 TI - Induction of CYP3A isoforms in cultured precision-cut human liver slices. AB - 1. The effect of rifampicin on cytochrome P450 isoforms in the CYP1A and CYP3A subfamilies has been studied in 72-h cultured precision-cut human liver slices. 2. In cultured human liver slices 50 microM rifampicin induced testosterone 6 beta-hydroxylase activity, but had no effect on 7-ethoxyresorufin O-deethylase and 7-methoxyresorufin O-demethylase activities. 3. Western immunoblotting of liver slice microsomes was performed with antibodies to rat CYP1A2 and human CYP3A4. Compared with control (dimethyl sulphoxide only treated) liver slice microsomes, rifampicin increased levels of CYP3A4 but had no effect on CYP1A2. 4. These results demonstrate that rifampicin induces CYP3A isoforms, but not CYP1A2, in cultured human liver slices. Some variability in the magnitude of induction by rifampicin was observed in the six human liver samples examined. 5. These results demonstrate that cultured human liver slices may be used to evaluate the effects of xenobiotics on CYP3A isoforms. PMID- 9413921 TI - NMR spectroscopic studies on the metabolism and futile deacetylation of phenacetin in the rat. AB - 1. 1H-NMR spectroscopy of urine was used to determine the % deacetylation and re acetylation of 2H-labelled (in the acetyl) phenacetin metabolites in the rat. 2. Male Sprague-Dawley rats were each dosed with either phenacetin or phenacetin C2H3 at 50 mg kg-1. The total urinary recoveries for phenacetin and phenacetin C2H3 were 47.6 +/- 16.7 and 50.1 +/- 16.2% respectively (not significantly different, p > 0.05). Paracetamol sulphate and glucuronide are the major urinary metabolites of both protio and deuteriophenacetin. 3. The futile deacetylation given by the urinary recovery of protio-acetyl metabolites of phenacetin-C2H3 was 29.6 +/- 0.9% for paracetamol sulphate and 36.6 +/- 3.1% for paracetamol glucuronide. These observations demonstrate a high level of futile deacetylation in the paracetamol conjugates formed by metabolism of phenacetin-C2H3 and this may indicate a high metabolic flux through the nephrotoxic intermediate 4 aminophenol. 4. The level of futile deacetylation for phenacetin was significantly higher than that found previously in studies of labelled paracetamol in rat or man, and may be important in understanding the higher nephrotoxicity of phenacetin as compared with paracetamol. PMID- 9413922 TI - Metabolic disposition of [14C]-trimethylamine N-oxide in rat: variation with dose and route of administration. AB - 1. Urine was the major route of excretion of radioactivity (95% dose in 0-24 h) following the oral, intravenous or intraperitoneal administration of [14C] trimethylamine N-oxide dihydrate (1 mmol/kg body wt) to the adult male Wistar rat. A further 3-4% was voided in the urine during 24-72 h. Only fractional amounts were detected in the faeces, or were retained within tissues 3 days after administration. 2. Biliary secretion of radioactivity was insignificant (0.18% in 0-4 h) but larger amounts were secreted directly into the lumen of the gastrointestinal tract, especially the small intestine (2.6% in 0-1 h). 3. The only radioactive compounds identified in the urine were trimethylamine N-oxide and dimethylamine. Larger amounts of dimethylamine were excreted following oral administration (10%) as opposed to intravenous (2.5%) or intraperitoneal (1.5%) input. This production of dimethylamine occurred over a 100-fold oral trimethylamine N-oxide dose range (0.3-30 mmol/kg body wt). Incubation of trimethylamine N-oxide with gut contents (especially colon and rectum) led to the formation of dimethylamine. PMID- 9413923 TI - Receptors for natriuretic peptides in adrenal chromaffin cells. AB - Atrial, brain, and C-type natriuretic peptides of the atrial natriuretic peptide family are present in adrenal chromaffin cells, and are secreted with catecholamines by exocytosis. These peptides modulate the physiological functions of the cells such as synthesis and secretion of catecholamines in an autocrine manner interacting with natriuretic peptide receptors. PMID- 9413924 TI - Antisense oligonucleotides: is the glass half full or half empty? AB - Antisense oligonucleotides are widely used as tools to explore the pharmacological effects of inhibiting expression of a selected gene product. In addition, they are being investigated as therapeutic agents for the treatment of viral infections, cancers, and inflammatory disorders. Proof that the pharmacological effects produced by the oligonucleotides are attributable to an antisense mechanism of action requires careful experimentation. Central to this problem is the finding that oligonucleotides are capable of interacting with and modulating function of specific proteins in both a sequence-independent and dependent manner. Despite these undesired interactions, it has been possible to demonstrate that oligonucleotides are capable of binding to a specific RNA in cultured cells, or within tissues, resulting in selective reduction of the targeted gene product and pharmacological activity. In general, these oligonucleotides were identified after a selection process in which multiple oligonucleotides targeting different regions on the RNA were evaluated for direct inhibition of targeted gene product, resulting in the identification of a potent and selective oligonucleotide. Similar to other drug-receptor interactions, selection of the most potent inhibitor results in an increase in the signal-to noise ratio, yielding increased confidence that activity observed is the result of a desired effect of the inhibitor. With careful selection, proper controls, and careful dose-response curves it is possible to utilize antisense oligonucleotides as effective research tools and potentially as therapeutic agents. PMID- 9413925 TI - Decreased release of nitric oxide (NO) by alveolar macrophages after in vivo loading of rats with either iron or ethanol. AB - Alveolar macrophages were isolated by pulmonary lavage from rats which had been either chronically overloaded with iron by intraperitoneal injections of iron dextran for four weeks, or rendered alcoholic by administration of increasing concentrations of alcohol vapour, also for four weeks. Although the hepatic iron content increased in both groups of animals, only the macrophages isolated from the iron-loaded animals showed a significant increase in iron content (P = < 0.05). Furthermore, in these macrophages there was a significant increase in oxidative tone as demonstrated by a six fold increase in superoxide dismutase activity. In both the iron-loaded and chronically alcoholised macrophages, there was a significant diminution in nitric oxide release after stimulation with lipopolysaccharide and/or interferon-gamma, which impaired the ability of both of these groups of macrophages to inhibit the germination of spores from the fungus Rhizopus, a nitric oxide-dependent process. Such an alteration in nitric oxide release reduces the macrophage's microbicidal activity. PMID- 9413926 TI - Equilibrium binding of anthracycline cytostatics to serum albumin and small unilamellar phospholipid vesicles as measured by gel filtration. AB - A Sephadex G-200 gel filtration method was used to measure directly the equilibrium binding of five important anthracycline analogs to serum albumin. The order of the overall binding constant (K) in a 150 mM NaCl, 20 mM Hepes buffer (pH 7.45) was doxorubicin < daunorubicin < 4-demethoxydaunorubicin approximately 13-dihydro-4'-deoxy-4'-iododoxorubicin < 4'-deoxy-4'-iododoxorubicin for human serum albumin (K = 2.67 +/- 0.07 mM(-1) to 24.5 +/- 3.1 mM[-1]) and bovine serum albumin (K = 1.36 +/- 0.25 mM(-1) to 48.4 +/- 5.2 mM[-1]). Data were given on the pH-dependence of K. The anthracycline-albumin association reaction was compared with measurements of drug partitioning into unilamellar phospholipid membranes and octanol. The results provide important new data required for a systematic kinetic analysis of anthracycline transport in tumor cells under serum conditions in a biological system. PMID- 9413927 TI - Regional differences in the metabolism of Tyr-MIF-1 and Tyr-W-MIF-1 by rat brain mitochondria. AB - Tyr-MIF-1 (Tyr-Pro-Leu-Gly-NH2) and Tyr-W-MIF-1 (Tyr-Pro-Trp-Gly-NH2) are endogenous neuropeptides with opiate modulating and other CNS effects. After incubation of the tritiated tetrapeptides with fractions of tissue from different areas of rat brain, formation of the metabolites was determined by HPLC. Marked regional differences in degradation were found for both peptides. The metabolism of Tyr-MIF-1, resulting in the formation of the biologically active MIF-1 (Pro Leu-Gly-NH2), was greater in the mitochondrial than in the synaptosomal fractions. In the mitochondrial fraction, about twice as much MIF-1 was formed in brain cortex than in striatum, diencephalon, or midbrain/pons medulla. These results, showing differential metabolism in various areas of the brain, indicate another means for regulation of the concentrations of neuropeptides. PMID- 9413928 TI - Evidence of a coupled mechanism between monoamine oxidase and peroxidase in the metabolism of tyramine by rat intestinal mitochondria. AB - The relationship between monoamine oxidase (EC 1.4.3.4; MAO) and peroxidase (EC 1.11.1.7; POD) in the metabolism of tyramine was investigated using the crude mitochondrial fraction of rat intestine. When tyramine was incubated with mitochondria, the formation of the peroxidase-catalysed oxidation product, 2,2' dihydroxy-5,5'-bis(ethylamino)diphenyl (dityramine), identified by mass spectrometric analysis, was monitored spectrophotometrically. After an initial lag time, the formation rate of dityramine was linear up to 2 hr, amounting to 17 nmol x hr(-1) x mg protein(-1). A similar value was found for the oxidative deamination of tyramine catalysed by intestinal MAO. Either 10(-3) M clorgyline or 10(-3) M NaCN suppressed this reaction by completely inhibiting MAO or POD, respectively. In the former case, however, addition of H2O2 to the incubation mixture promptly started the reaction. Selective inhibition of MAO-A and MAO-B was achieved with 3 x 10(-7) M clorgyline and 3 x 10(-7) M deprenyl, respectively, and the formation rate of dityramine decreased in a corresponding manner. Preincubation with histamine or spermidine reduced the lag time without affecting the steady-state reaction rate. Higher levels of dityramine were also detected in vivo in rat intestine after oral administration of tyramine. These results indicate that the peroxidase-dependent metabolism of tyramine in the gut may be driven by H2O2 produced by MAO activities and that MAO-A is mainly responsible for this process, as well as for the oxidative deamination of tyramine. PMID- 9413929 TI - Altered cyclic AMP-dependent human chorionic gonadotropin production in cultured human placental trophoblasts exposed to ethanol. AB - Chronic ethanol abuse during pregnancy can cause fetal injury, including the fetal alcohol syndrome (FAS). A contributing factor in this fetal injury may be the effect of ethanol on placental function. Previous studies have shown that ethanol treatment increases human chorionic gonadotropin (hCG) production by cultured human placental trophoblasts. In this study, we demonstrated that the stimulation of hCG production correlates with the ethanol concentration. Ethanol treatment enhanced intracellular adenosine 3':5'-cyclic monophosphate (cAMP) levels in response to either cholera toxin (CTX) or forskolin (FSK). Moreover, basal (i.e. unstimulated) cAMP levels were increased at 2 hr of ethanol exposure. However, this effect did not persist throughout the 24-hr incubation period. Therefore, ethanol treatment appears to induce increased hCG production, secondary to enhanced basal or stimulated cAMP production. The effect of ethanol was not associated with changes in Gs or Gi2 expression, as determined by northern blot and western blot analyses. In plasma membrane preparations from ethanol-treated cells, cAMP production was higher in response to Mn2+, a direct stimulator of adenylyl cyclase. Inclusion of Rp-cAMP, a protein kinase A inhibitor, eliminated the ethanol effect on hCG production. Treatment of cells with 8-Br-cAMP stimulated hCG production, but there was no difference between the ethanol-naive control and the ethanol-treated cells. These data suggest that ethanol treatment increases in vitro hCG production in human placental trophoblasts by enhancing cAMP production. Ethanol treatment appears to increase trophoblast adenylyl cyclase activity. PMID- 9413930 TI - Effect of aspirin on induction of apoptosis in HT-29 human colon adenocarcinoma cells. AB - Aspirin (ASA) and other nonsteroidal anti-inflammatory drugs (NSAIDs) inhibit colorectal tumorigenesis. Apoptosis is a critical determinant of tissue mass homeostasis and may play a role in carcinogenesis. We studied the effect of ASA on the survival of a human colon cancer cell line using more sensitive methods than we had applied previously. ASA induced apoptosis in HT-29 colon adenocarcinoma cells at concentrations > or =1 mM as established by: (a) morphological changes consistent with apoptosis in cells examined by fluorescence microscopy and semi-thin cell sections, and (b) DNA strand breaks: 45% of the cells were TdT-mediated dUTP nick end labeling (TUNEL) positive at 3 mM at 72 hr, and 70% were positive by the comet assay. Electron microscopy also confirmed the induction of apoptosis by ASA. ASA-induced apoptosis was not associated with: (a) a ladder pattern on genomic DNA electrophoresis, or (b) a subdiploid peak on flow cytometry. Apoptotic bodies were virtually absent on standard morphological assessments and only a few were detected on semi-thin sections. For the above reasons, this apoptosis induced by ASA is "atypical," and the unusual features of ASA-induced apoptosis, besides their taxonomic value, may offer clues to the mechanisms that control the process of apoptosis or perhaps the cancer chemopreventive properties of this compound. These findings demonstrate that ASA induces apoptosis in human colon cancer cells, bolstering the hypothesis that apoptosis may be a mechanism by which NSAIDs inhibit colon carcinogenesis. These findings should be examined in animal and/or clinical research studies in vivo. PMID- 9413931 TI - Rat beta-adrenergic receptor kinases 1 and 2 in mouse neuroblastoma X rat glioma NG 108-15 hybrid cells. AB - Beta-adrenergic receptor kinase (betaARK, EC 2.7.1.-) has been implicated in the phosphorylation of G protein-coupled receptors, including opioid receptors. Since delta-opioid receptors of mouse neuroblastoma x rat glioma hybrid cells (NG 108 15) desensitize upon activation, this investigation was designed to find out whether NG 108-15 cells contain betaARK activity. Using the reverse transcription polymerase chain reaction technique, we identified two mRNAs, one coding for rat betaARK1 and the other for rat betaARK2. No hint was found for the presence of mouse betaARK. Examining the cytosolic betaARK activity in these hybrid cells using rhodopsin as substrate, we found a strict functional dependence on the presence of exogenous G protein subunit Gbetagamma. This relationship reflects a characteristic for betaARK1 and 2 out of the known G protein-coupled receptor kinases. Finally, highly purified recombinant betaARK1 proved active to phosphorylate enriched delta-opioid receptor preparations in an opioid agonist dependent manner. The results reported here provide the basis to study more closely the molecular function of G protein-coupled receptor kinases in a cell line (NG 108-15) most frequently used to investigate acute and chronic opioid actions. PMID- 9413932 TI - Inhibitory effect of genistein on bone resorption in tissue culture. AB - The effect of genistein on bone resorption in vitro was investigated. Femoral metaphyseal tissues obtained from elderly female rats were cultured for 48 hr in Dulbecco's modified Eagle's medium (high glucose, 4.5%) supplemented with antibiotics and bovine serum albumin. The experimental cultures contained 10(-7) to 10(-3) M genistein. The bone-resorbing factors parathyroid hormone (1-34) (PTH; 10(-7) M), prostaglandin E2 (PGE2; 10(-5) M), and lipopolysaccharide ( 10 microg/mL) caused a significant decrease in bone calcium content. The decrease in bone calcium content induced by bone-resorbing factors was inhibited completely by genistein (10(-7) to 10(-5) M). In addition, this isoflavonoid (10(-5) M) completely inhibited the PTH (10(-7) M)- or PGE2 (10(-5) M)-induced increase in medium glucose consumption and lactic acid production by bone tissues. Moreover, genistein (10(-5) M) blocked both PTH (10(-7) M)-increased acid phosphatase and decreased alkaline phosphatase activities of bone tissues. The inhibitory effect of genistein (10(-5) M) on PTH (10(-7) M)-stimulated bone resorption was clearly prevented by the presence of 10(-6) M tamoxifen, an anti-estrogen reagent. Genistein (10(-5) M) did not further enhance the inhibitory effect of estrogen (10(-9) M) on PTH-stimulated bone resorption. These findings indicate that genistein has a direct inhibitory effect on bone resorption in tissue culture in vitro. PMID- 9413933 TI - Stimulation of nitric oxide synthase during oxidative endothelial cell injury. AB - The purpose of this study was to determine changes in nitric oxide synthase (NOS) activity during the process of lethal oxidative cell injury following H2O2 treatment of endothelial cells. NOS activity was determined by measuring the conversion of [3H]arginine ([3H]Arg) to [3H]citrulline ([3H]Cit). Cell death was assessed by measuring the release of intracellular lactate dehydrogenase (LDH). Moreover, cell death and changes in cytosolic free Ca2+ (Ca(i)2+) were measured simultaneously using a confocal laser scanning system, and propidium iodide and fluo-3 as fluorescent indicators, respectively. Treatment with H2O2 (125-1000 microM) concentration dependently increased L-Cit formation from L-Arg, and a peak was obtained at 90 min after the addition of 500 or 1000 microM H2O2. The H2O2-induced increase in L-Cit formation was blocked completely by N(G)-nitro-L arginine (L-NNA) or N(G)-methyl-L-arginine (L-NMA), both inhibitors of NOS. LDH release from endothelial cells was evoked from 120 min after the addition of H2O2 (125-1000 microM) in a concentration-dependent manner. Moreover, H2O2 increased Ca(i)2+ before cell death, and addition of Ca2+ chelator inhibited both the increase in L-Cit formation and LDH release by H2O2. The H2O2-induced LDH release was reduced by L-NNA, but not by L-NMA. These results suggest that H2O2 treatment of endothelial cells increases Ca(i)2+ before cell death, and stimulates NOS activity. The activation of NOS may be involved in oxidative endothelial cell death. PMID- 9413934 TI - Ubiquinone-0 (2,3-dimethoxy-5-methyl-1,4-benzoquinone) as effective catalyzer of ascorbate and epinephrine oxidation and damager of neuroblastoma cells. AB - The kinetics of ascorbate (AscH ) and epinephrine (EP) oxidation in the presence of 2,3-dimethoxy-5-methyl-1,4-benzoquinone (UQ) were studied in 0.05 M phosphate buffer, pH 7.4, at 37 degrees C by using a Clark electrode and ESR techniques. UQ at nanomolar concentrations displayed a pronounced catalytic effect on AscH oxidation which exceeded that of all reported organic catalysts tested in this system. The process was accompanied by the intensive oxygen consumption and increase in the steady-state concentration of the ascorbyl radical Asc.-. The rate of oxygen consumption (R[OX]) was maximal at the moment of reagent mixing ((R[OX]0) and then reduced over a few minutes until a steady-state level ((R[OX])SS) was achieved. (R[OX])0 was found to be proportional to [UQ][AscH-] without regard to the concentrations of the individual reagents; (R[OX])SS was directly related to [UQ] at a given concentration of AscH-. The difference between (R[OX])0 and (R[OX])SS decreased as [AscH-] decreased. The presence of a lipid phase (sodium dodecylsulphate micelles) only moderately decreased UQ activity as a catalyst of AscH- oxidation. Adding micromolar concentrations of UQ induced the acceleration of EP autoxidation. The capability of UQ to catalyze the oxidation of EP exceeded by approximately 25 times that of adrenochrome, a quinoid product of EP oxidation. These catalytic properties of UQ allowed us to predict its pronounced cytotoxicity, especially in the presence of AscH- and to cells of the sympathetic nervous system which are rich in catecholamines. This possibility was confirmed by experiments with human neuroblastoma cells in culture. The capability of UQ to injure neuroblastoma cell line SK-N-SH exceeded that of well-known neurotoxic agents 6-hydroxydopamine and menadione. PMID- 9413935 TI - Pharmacological effect of tetracyclines on proteoglycanases from interleukin-1 treated articular cartilage. AB - Based on previous in vivo and in situ studies showing that tetracyclines possess antidegenerative effects on cartilage in conjunction with a reduced proteoglycan (PG) loss from the extracellular matrix, we investigated the effects of doxycycline, minocycline and tetracycline on the degradation and biosynthesis of PGs by bovine articular cartilage explants, both in vitro and in situ. Doxycycline, minocycline and tetracycline dose dependently, although weakly, inhibited PG degrading matrix metalloproteinases (MMPs) in vitro, when tested at concentrations ranging from 1 to 100 microM. Ro 31-4724 proved to be a potent inhibitor of MMP proteoglycanases (IC50 value 1.5 nM). Only at a concentration of 100 microM did doxycycline and minocycline significantly inhibit the interleukin 1 (IL-1)-induced augmentation of PG loss from cartilage explants into the nutrient media. The tetracyclines did not modulate the IL-1-mediated reduced aggregability of PGs, whereas 10 microM Ro 31-4724 partially restored the aggregability of PGs ex vivo. Tetracycline even at this high concentration was ineffective. Compared to the effects of the MMP inhibitor Ro 31-4724, treatment with tetracyclines at therapeutic serum levels of 1 or 10 microM was minimal, with little or no effect on cartilage proteoglycanases and PG biosynthesis. In our experiments, tetracyclines and Ro 31-4724 at doses evaluated had no cytotoxic effects on chondrocytes. PMID- 9413936 TI - Catechol estrogens as inhibitors of leukotriene synthesis. AB - Estrogens have a beneficial effect on atherosclerosis and osteoporosis after menopause, but their exact mechanism of action is still unknown. The aim of the present study was to investigate the effects of estradiol and its metabolites catechol estrogens on arachidonic acid metabolism in vitro. Estradiol had no effect on arachidonic acid metabolism up to 33 microM in A23187-stimulated human whole blood. All catechol estrogens (2-hydroxyestradiol, 2-hydroxyestrone, 4 hydroxyestradiol and 4-hydroxyestrone) had similar kinds of actions on arachidonic acid metabolism, being over ten times more potent inhibitors of leukotriene synthesis (IC50 values 0.044-0.16 microM) than thromboxane (IC50 values 0.99-2.1 microM) and prostaglandin E2 synthesis (IC50 values 0.84-5.5 microM). It is suggested that some of the protective actions of estrogens--e.g., on atherosclerosis and osteoporosis--may be related to the inhibition of leukotriene synthesis by catechol estrogens. PMID- 9413937 TI - High-contact paternal occupations, infection and childhood leukaemia: five studies of unusual population-mixing of adults. AB - The hypothesis has been tested that, among excesses of childhood leukaemia associated with extreme population-mixing, the incidence is higher for the children of men in occupations involving contact with many individuals (particularly children), as noted in certain childhood infections. Data on childhood leukaemia were examined from five previous studies of the author in which significant excesses had been found associated with population-mixing involving adults. Occupational titles were categorized according to the estimated level of work contacts as medium, high, very high or indeterminate. Occupations involving frequent contact with children were categorized as having a very high contact level given the high frequency of exposure to the infection postulated as underlying childhood leukaemia. There was a significant positive trend (P < 0.001) in childhood leukaemia risk at ages 0-14 years across the occupational contact categories from the reference group (comprising the medium and low plus indeterminate categories) through high to very high (i.e. high-child) contact categories in the combined data from the author's five studies of adult population-mixing; this significant trend also applied at ages 0-4 (P < 0.001) and 5-14 (P < 0.01) years. The excess in the high category was mainly because of paternal occupations connected with the construction industry and transport, suggesting a broader definition of the 'very high' contact category. No sign of these excesses was found in a limited examination of the question outside areas of population-mixing using mortality data for childhood leukaemia in the general population of England and Wales. The findings represent the first individual based support for infection underlying childhood leukaemia that is promoted by population-mixing, as well as further support for the role of adults in transmission of the infection. PMID- 9413938 TI - Absence of mutations in the ATM gene in breast cancer patients with severe responses to radiotherapy. AB - The effectiveness of cancer radiotherapy is compromised by the small proportion (approximately 5%) of patients who sustain severe normal tissue damage after standard radiotherapy treatments. Predictive tests are required to identify these highly radiosensitive cases. Patients with the rare, recessively inherited, cancer-prone syndrome ataxia-telangiectasia (A-T) sustain extremely severe normal tissue necrosis after radiotherapy and their cultured cells are also highly radiosensitive. Clinically normal carriers (heterozygotes) of the A-T gene have an increased risk of breast cancer, account for approximately 4% of all breast cancer cases and show a modest increase in cellular radiosensitivity in vitro. It has been suggested that a substantial proportion of highly radiosensitive (HR) breast cancer patients may be A-T heterozygotes, and that screening for mutations in the A-T gene could be used as a predictive test. We have tested this hypothesis in a group of cancer patients who showed adverse reactions to radiotherapy. Sixteen HR breast cancer patients showing mainly acute reactions (and seven HR patients with other cancers) were tested for ATM mutations using the restriction endonuclease fingerprinting assay. No mutations typical of those found in obligate A-T heterozygotes were detected. If the estimate that 4% of breast cancer cases are A-T gene carriers is correct, then ATM mutations do not confer clinical radiosensitivity. These early results suggest that screening for ATM mutations in cancer patients may not be of value in predicting adverse reactions. PMID- 9413939 TI - Exclusion of BBC1 and CMAR as candidate breast tumour-suppressor genes. AB - Loss of heterozygosity (LOH) on chromosome arm 16q occurs in 48-65% of breast tumours. One small region of overlap is located at 16q24.3. Two genes located in this region, the cellular adhesion regulatory molecule (CMAR) and the breast basic conserved gene (BBC1), are plausible candidate tumour-suppressor genes. Mutational analysis of the retained copy of these genes has been performed by direct sequencing in a selected set of breast tumours that show LOH at 16q24.3 but not at other regions on chromosome arm 16q. In CMAR no other alterations than the previously described 4-bp insertion of CACA at nucleotide 241 could be detected, which was also present in constitutional DNA of the same patients. This polymorphism occurs homozygously in germline DNA of normal individuals and breast cancer patients. LOH analysis at this locus shows no preferential loss of a particular variant of the 241 polymorphism. In the BBC1 gene, three different alterations were found, but only one resulted in an amino acid substitution. This is a known polymorphism, however, also appearing in germline DNA. The absence of tumour-specific mutations in CMAR and BBC1 in this selected series of breast tumours implies that another gene at 16q24.3 must be the tumour-suppressor gene that is the target for LOH in breast cancer. PMID- 9413941 TI - Loss of heterozygosity at the mannose 6-phosphate insulin-like growth factor 2 receptor gene correlates with poor differentiation in early breast carcinomas. AB - Chromosome 6q has been shown to be one of the most frequent sites for allelic loss in human breast cancer. The mannose 6-phosphate/insulin-like growth factor 2 receptor (IGF2R) gene, which maps to chromosome 6q26-27, functions in the activation of TGF-beta1, a potent growth inhibitor for most cell types, the degradation of the mitogen IGF2 and the intracellular trafficking of lysosomal enzymes. Loss of heterozygosity (LOH) at the IGF2R locus with mutations in the remaining allele have been reported in liver cancers and recently in two high grade cases of ductal carcinoma in situ of the breast. We have sought to confirm that allelic loss of IGF2R is an early event in the aetiology of breast cancer by screening a group of 'early' lesions for LOH at a polymorphic microsatellite marker within the IGF2R gene using polymerase chain reaction (PCR). Several microdissected tumour foci were analysed for each of 40 mammographically detected invasive carcinomas and 22 cases of pure ductal carcinoma in situ (DCIS). None of 25 (62.5%) informative early invasive carcinomas showed any evidence of LOH. This group comprised predominantly of well- to moderately differentiated cases (95%). However, 4 out of 18 informative DCIS cases (22%) showed clear evidence of LOH. Three of these were poorly differentiated (high-grade) lesions. These data suggest that loss of heterozygosity at the IGF2R gene is associated with poor differentiation at this early stage of breast cancer development and progression. PMID- 9413940 TI - Up-regulation of p21WAF1 expression in myeloid cells is activated by the protein kinase C pathway. AB - Phorbol-12-myristate-13-acetate (PMA) induces p21WAF-1 expression in human myeloid leukaemic HL-60 cells. We show that this induction is specifically mediated by protein kinase C (PKC). In addition, the PKC inhibitor Ro 31-8220 with predominant PKC-alpha isoform specificity almost completely inhibited PMA induced up-regulation of p21WAF1 in HL-60 cells as well as in the myelomonocytic leukaemic U937 cells. Pretreatment of HL-60 cells with Ro 31-8220 also inhibited PMA-induced activation of c-raf-1, a known PKC alpha target. In the phorbol ester tolerant HL-60 subline (PET) with PKC-beta isoform deficiency PMA or bryostatin-1 induced p21WAF1 expression, but to a lesser extent than in wild-type HL-60 cells. In PET cells, Ro 31-8220 also inhibited PMA and bryostatin-1-induced up regulation of p21WAF1 expression. Our findings indicate that at least in HL-60 cells up-regulation of p21WAF-1 is specifically activated by PKC. We suggest that PKC isoforms other than beta, presumably the PKC-alpha isoform, are involved in this process. PMID- 9413942 TI - Transcription of the gene encoding melanoma-associated antigen gp100 in tissues and cell lines other than those of the melanocytic lineage. AB - The expression of the gp100 antigen is generally thought to be confined to cells of the melanocytic lineage, which makes the protein a suitable melanoma-specific marker. Strikingly, after screening a panel of normal tissues, tumour samples and cell lines of non-melanocytic origin, we found transcripts encoding gp100 in virtually every tissue and cell line tested. In contrast, tyrosinase and MART 1/MelanA transcripts were detected only in cells of the melanocytic lineage. However, no gp100 protein could be detected by either Western blotting or cytotoxicity assays. Therefore, at the protein level, gp100 remains exclusive for cells of melanocytic origin despite its transcription in many cell types. The major implication of this finding is that screening of patient material for gp100 expression should preferrably be performed by antibody staining. Reverse transcriptase polymerase chain reaction (RT-PCR) can be employed, provided that it is performed in a tightly controlled, semiquantitative setting. PMID- 9413943 TI - Commonly deleted region on the long arm of chromosome 7 in differentiated adenocarcinoma of the stomach. AB - Loss of heterozygosity (LOH) at several chromosomal loci is a common event in human malignancies. Frequent LOH on the long arm of chromosome 7 has been reported in various human malignancies, and investigators have identified the most common site of LOH as 7q31.1. We have identified ten chromosomal loci, including chromosome 7q, that have been shown by previous allelotype study to be sites of frequent LOH in differentiated adenocarcinoma of the stomach. In the present study, we performed a polymerase chain reaction (PCR) microsatellite analysis to define the common deleted region on 7q, using 14 polymorphic microsatellite markers in matched tumour and non-tumour DNAs from 53 patients with primary gastric carcinoma of the differentiated type. LOH at any locus on 7q occurred in 34% (18 out of 53) of the tumours. Although many tumours exhibited total or large interstitial deletions, we determined the smallest common deleted region to be at D7S480 (7q31.1). This is identical to the region identified for other human malignancies. These observations indicate that a putative tumour suppressor gene at 7q31.1 may be involved in the pathogenesis of differentiated adenocarcinoma of the stomach. PMID- 9413944 TI - Trafficking of activated lymphocytes into the RENCA tumour microcirculation in vivo in mice. AB - The aim of the study was to establish a model of tumour microcirculation in vivo using the murine renal cell carcinoma cell line (RENCA) implanted into the mouse cremaster muscle, and subsequently to investigate the trafficking of syngeneic lymphocyte subpopulations into both the RENCA tumour and the surrounding normal cremaster muscle microcirculation. We have demonstrated that RENCA tumour cells, at a dose of 1.5 x 10(5) per 30 microl injected into the cremaster muscle, reproducibly produced a vascularized tumour suitable for in vivo microscopy at 10 14 days. Injection of fluorescently labelled effector cells (1 x 10(6)) including naive splenocytes, T-cell enriched populations and ex vivo interleukin 2 (IL-2) activated splenocytes all migrated to and flowed through both the tumour and the normal microcirculation, with negligible adhesion. However, we observed the selective recruitment, localization and arrest of IL-2-activated splenocytes (P < 0.05) into the tumour microcirculation, and the subsequent extravasation of cells into the tumour intestitium in some instances. This did not occur with the other effector cells. We also observed the absence of leucocyte rolling in the tumour microcirculation, suggesting an impairment in adhesion molecule expression on the tumour endothelium. We have therefore established the potential of this model for defining further effector cell-tumour-endothelium interactions. PMID- 9413945 TI - The relationship between intrinsic thymidylate synthase expression and sensitivity to THYMITAQ in human leukaemia and colorectal carcinoma cell lines. AB - Thymidylate synthase (TS) expression has been characterized for a panel of eight human colorectal carcinoma and five human leukaemia cell lines, to relate differences in intrinsic TS activity, protein and mRNA levels to growth inhibition caused by continuous exposure to THYMITAQ, a specific non-classical antifolate TS inhibitor. Although a 20-fold variation in sensitivity to THYMITAQ was found within the colorectal cell line panel (IC50 0.12-2.7 microM), sensitivity was not related to TS activity, TS protein or TS mRNA levels. For the leukaemic cell lines, only a twofold range in sensitivity to THYMITAQ was observed (IC50 0.87-2.3 microM), and this did not correlate with TS activity, TS protein or TS mRNA levels. Across all of the cell lines, TS activity was linearly related to TS protein levels (r2 = 0.87, P < 0.0001). However, for both the colorectal and leukaemia cell line panels, no relationship was found between TS mRNA/18S rRNA ratios and either TS activity or TS protein, consistent with the importance of post-transcriptional mechanisms in regulating TS activity. Two of the colorectal cell lines (BE and HCT116) and one of the human leukaemic cell lines (HL60), were intrinsically resistant to THYMITAQ (IC50 > 2 microM) in the absence of TS overexpression, suggesting that, subsequent to TS inhibition, events such as DNA repair and tolerance to apoptotic stimuli are also important determinants of sensitivity to THYMITAQ. PMID- 9413946 TI - Production of tumour necrosis factor-alpha by cultured human peripheral blood leucocytes in response to the anti-tumour agent 5,6-dimethylxanthenone-4-acetic acid (NSC 640488). AB - The investigative anti-tumour agent 5,6-dimethylxanthenonone-4-acetic acid (DMXAA, NSC 640488), developed in this laboratory as an improved analogue of flavone acetic acid (FAA, NSC 347512), is currently in clinical trial. The ability of DMXAA to up-regulate tumour necrosis factor (TNF) mRNA and protein synthesis in cultured human peripheral blood leucocytes (HPBLs) has been investigated and compared with that of flavone acetic acid (FAA) and of bacterial lipopolysaccharide (LPS). Human peripheral blood leucocytes were isolated from buffy coats obtained from a blood transfusion centre and also from blood samples from laboratory volunteers. At a concentration of 400 microg ml(-1) and an incubation time of 2 h, DMXAA up-regulated mRNA synthesis in six of eight individuals tested, as measured by Northern blotting. The degree of up-regulation varied in different individuals from one to nine times that of control levels. In contrast, FAA caused no induction above that of control levels and in some cases suppressed expression relative to controls, extending previous data that DMXAA but not FAA up-regulates TNF mRNA in the human HL-60 tumour cell line. At the same concentration but with longer incubation times (6-12 h), DMXAA induced increases in TNF protein in 11 of 15 samples of HPBLs from buffy coats and also in 11 of 15 samples of HPBLs from volunteers, as measured by cytotoxicity assays with L929 cells. FAA caused no increase in TNF protein, while LPS induced TNF to approximately 20-fold higher levels than did DMXAA. Considerable heterogeneity of response was observed with both sources of HPBLs, and there was little or no correlation between the extent of TNF induction by DMXAA and LPS in individual samples. In vitro analysis of the response of human peripheral blood leucocytes to DMXAA may be a useful test in clinical trials of agents such as DMXAA. PMID- 9413947 TI - Thrombin has a bimodal effect on glioma cell growth. AB - Using rat glioma C6 cells as a model, we have found a bimodal effect of alpha thrombin on cell growth. In C6 cells treated with alpha-thrombin at concentrations from 0.02 nM to 1.0 nM, inhibition of cell proliferation was noted. Because the thrombin receptor agonist peptide TRAP-6 also induced inhibition of cell proliferation and the thrombin receptor antagonist peptide T1 prevented the inhibitory effect of alpha-thrombin on C6 glioma cell growth, thrombin receptor involvement in antiproliferative action of alpha-thrombin in C6 glioma cells is highly likely. However, stimulation of cell proliferation observed when C6 cells were treated with alpha-thrombin at higher doses (> 1.0 nM) seems to be mediated by as yet undefined thrombin receptor-independent biochemical mechanisms. PMID- 9413949 TI - Multiple simultaneous gastric carcinomas. AB - A total of 1664 patients with gastric cancer were examined to evaluate the rate of multiple synchronous primary tumours. In cases of multiple synchronous cancer (MSC), the tumours were analysed immunohistochemically for their expression pattern of p53, c-erbB2, ras, E-cadherin and proliferative activity. Multiple synchronous gastric carcinomas (MSCs) were observed in 61 out of 1664 patients (3.7%), with a total of 134 carcinomas. In our series, early carcinoma was observed more frequently in MSC than in solitary cancers. The comparison of tumour stage in MSC and solitary tumours revealed that multiple early gastric cancers were significantly more often of type I (protruded type) and IIa (superficial elevated type) than solitary early cancer. Multiple advanced carcinomas were more often of a lower pT category than solitary advanced gastric cancer. Performing immunohistochemistry for p53, c-erbB2 and ras in 134 tumours with MSCs, we observed positivity rates of 33%, 59% and 87% respectively. In 43 patients, the multiple tumours in each individual patient demonstrated an identical status of p53 and c-erbB2, and in 42 patients a similar pattern of E cadherin expression was observed. The proliferative index, determined by proliferating cell nuclear antigen (PCNA) immunolabelling, did not differ significantly between the MSC in each patient. Ras immunostaining was detected in 53 out of 61 patients, but also in metaplasia and regenerative hyperplasia in the specimens. In survival analysis, no difference was observed between patients with solitary or multiple early or advanced carcinomas. Our results suggest that in at least a high proportion of patients with gastric cancer multiple primary tumours arise from precancerous conditions leading to similar genetic alterations. PMID- 9413948 TI - Involvement of DT-diaphorase (EC 1.6.99.2) in the DNA cross-linking and sequence selectivity of the bioreductive anti-tumour agent EO9. AB - The chemistry of the mitomycin C-related drug indoloquinone EO9 would suggest that its mechanism of action is likely to involve DNA damage after reductive activation. The ability of this agent to induce DNA damage in intact cells has been examined using alkaline filter elution. After treatment with pharmacologically relevant concentrations of EO9, both DNA strand breaks and interstrand cross-links were detected in rat Walker tumour cells and human HT29 colon carcinoma cells. These cell lines express relatively high levels of DT diaphorase (NAD(P)H: quinone acceptor oxidoreductase), which is believed to be involved in EO9 activation. The extent of DNA damage was increased by approximately 30-fold under hypoxia in BE colon carcinoma cells that express non functional DT-diaphorase, but this dramatic hypoxia enhancement was not seen in HT-29 cells. These data are consistent with cytotoxicity studies that indicate that DT-diaphorase appears to be important in EO9 activation under aerobic conditions, but other enzymes may be more relevant under hypoxia. The involvement of DT-diaphorase in DNA damage induction was further investigated using cell-free assays. DNA cross-links were detectable in plasmid DNA co-incubated with EO9, cofactor and DT-diaphorase but not in the absence of this enzyme. In contrast, using a Taq polymerase stop assay, monofunctional alkylation was detected in plasmid DNA without metabolic activation, although the sequence selectivity was altered after reduction catalysed by DT-diaphorase. PMID- 9413950 TI - Colorectal carcinoma in Hong Kong: epidemiology and genetic mutations. AB - The incidence of colorectal carcinoma is rising at an alarming pace in Asian urban societies such as Hong Kong. Detailed examination of the epidemiological pattern and genetic mutation of colorectal cancer in the Hong Kong Chinese population is overdue. We compared the reported age incidence of colorectal carcinoma in Hong Kong with that of Scotland and other countries. Hong Kong showed a much higher incidence of colorectal carcinoma among the young age groups. By comparison with other countries, this raised incidence among the young appeared to be related to southern Chinese societies. The recent dramatic rise in colorectal cancer in Hong Kong was largely attributable to an increase in the over 50 years age group, while the young incidence remained unchanged. We also defined the mutation spectrum of p53 and Ki-ras in 67 unselected cases by direct DNA sequencing. Interestingly, insertion/deletion mutations in p53 from colorectal carcinoma in Hong Kong showed a significantly higher frequency (17.2%) than the Scottish data (0%) and the world database (6.6%), although the overall frequency of p53 mutation (43%) in Hong Kong was similar to others. The high incidence of colorectal carcinoma in young people and the raised proportion of frameshift mutations in p53 encourage further search for a genetic basis for susceptibility to this disease in the Hong Kong Chinese population. PMID- 9413951 TI - First clinical trial of cat soft-tissue sarcomas treatment by electrochemotherapy. AB - Electrochemotherapy combines bleomycin and local electric pulses that allow cell permeabilization and free access of bleomycin to its intracellular target. We report the first veterinarian clinical trial of electrochemotherapy in 12 cats with spontaneous large soft-tissue sarcomas that suffered relapse after treatment with conventional therapies. Permeabilizing electric pulses were delivered using external surface electrodes, as well as new needle-shaped electrodes that were designed to be inserted in tumours for more effective treatment of several centimetre-thick tumour nodules. The electric pulses were applied to the tumours several times from 4 to 15-30 min after a bolus intravenous injection of 0.5 mg kg(-1) bleomycin. Tolerance to treatment was excellent without general side effects. The cats showed local inflammatory reactions for a few days and disease stabilization lasted from 2 weeks to 7 months. One partial regression was observed, and the general absence of nodule volume decrease can be explained by local fibrotic reactions. Histological analysis of biopsies also revealed massive tumour cell death. The cats' lifespan increased (P<<0.001), with a mean survival time of 6.1 months (maximum 18 months) compared with 0.8 months (maximum 1.5 months) for a group of 11 untreated control cats displaying similar carcinological features. Electrochemotherapy is clearly effective as a salvage treatment for large spontaneous solid tumours in adverse clinical situations and this is promising for future applications. PMID- 9413952 TI - Central nervous system tolerance to boron neutron capture therapy with p boronophenylalanine. AB - A rat spinal cord model was used to evaluate the effects of boron neutron capture irradiation on the central nervous system (CNS), using a range of doses of the boron delivery agent p-boronophenylalanine (BPA). Three doses of BPA 700, 1000 and 1600 mg kg(-1) were used to establish the biodistribution of boron-10 (10B) in blood, spinal cord and brain over a 3-h period after intraperitoneal (i.p.) administration. At the lowest dose of BPA used, blood 10B levels remained relatively stable over the 3-h sampling period. With the two higher doses of BPA, blood 10B concentrations were greatest at 1 h after BPA administration, and thereafter exhibited a biphasic clearance profile. The largest decline in blood 10B levels occurred between 1 and 2 h after i.p. injection and was most pronounced (approximately 45%) in the highest BPA dose group. Considered overall, 10B concentrations were marginally lower in the spinal cord than in the brain. Levels of 10B in both of these organs showed a slow but progressive increase with time after administration of BPA. The 10B concentration ratio for blood relative to CNS tissue increased with BPA dosage and reached a peak value of approximately 10:1 in the highest BPA dose group, at 1 h after i.p. injection. However, at 3 h after injection the 10B concentration ratios had decreased to approximately 3:1 in all of the BPA dose groups. After irradiation with thermal neutrons in combination with BPA at blood 10B concentrations of approximately 42 and approximately 93 microg g(-1), myelopathy developed after latent intervals of 20.0 +/- 0.6 and 20.0 +/- 1.2 weeks respectively. ED50 values (+/- s.e.) for the incidence of myelopathy were calculated from probit-fitted curves, and were 17.5 +/- 0.7 and 25.0 +/- 0.6 Gy after irradiation with thermal neutrons at blood 10B levels of approximately 42 and approximately 93 microg g(-1) respectively. The compound biological effectiveness (CBE) factor values, estimated from these data, were 0.67 +/- 0.23 and 0.48 +/- 0.18 respectively. This compared with a previous estimate of 0.88 +/- 0.14 at a blood 10B concentration of approximately 19 microg g(-1). It was concluded that the value of the CBE factor was not influenced by the level of 10B in the blood, but by the blood:CNS 10B concentration ratio. In effect, the CBE factor decreases as the concentration ratio increases. Simulations using boron neutron capture therapy (BNCT) treatment planning software indicate a significant therapeutic advantage could be obtained in moving to higher BPA doses than those in current clinical use. PMID- 9413953 TI - The effects of treatment with chemotherapy on energy metabolism and inflammatory mediators in small-cell lung carcinoma. AB - A disturbed energy balance has been demonstrated in lung cancer patients. Both an enhanced resting energy expenditure (REE) and a decreased energy intake contribute to weight loss. Enhanced systemic levels of inflammatory mediators were found to be related to the enhanced REE in lung cancer. The aim of the present study was to investigate energy metabolism and systemic levels of inflammatory mediators in small-cell lung carcinoma (SCLC) patients before and after treatment with chemotherapy. Hypermetabolism and an enhanced inflammatory response have already been demonstrated in SCLC by our group before. Twelve newly diagnosed SCLC patients were consecutively included in the study. REE was measured by indirect calorimetry and body composition was determined by bioelectrical impedance (BIA) before and 1 month after treatment. To assess the inflammatory state the acute-phase proteins, C-reactive protein (CRP) and lipopolysaccharide-binding protein (LBP), both soluble tumour necrosis factor (TNF) receptors, (sTNF-R)-55 and sTNF-R75, and soluble intercellular adhesion molecule (sICAM)-1 were measured in plasma before and 1 month after treatment. CRP was assessed by turbidemetry, whereas the other inflammatory parameters were measured by enzyme-linked immunosorbent assay (ELISA). A significant reduction in REE was found irrespective of therapeutic outcome, whereas body weight and body composition remained stable. The acute-phase proteins CRP and LBP were reduced significantly after treatment with chemotherapy, whereas both sTNF receptors and sICAM-1 remained enhanced. No correlation, however, existed between the decrease in REE and the decrease in the acute-phase proteins. In conclusion, chemotherapeutic treatment attenuates the tumour-related metabolic derangements and acute-phase response. PMID- 9413954 TI - Continuous infusional topotecan in advanced breast and non-small-cell lung cancer: no evidence of increased efficacy. AB - Two open, phase II studies were performed to evaluate the activity and toxicity of infusional topotecan in patients with advanced non-small-cell lung carcinoma (NSCLC) and advanced breast cancer who had not received previous chemotherapy for metastatic disease. Twenty-five patients with an ECOG performance score < 2 were treated with infusional topotecan administered as a daily, continuous intravenous infusion starting at 0.6 mg m(-2) day(-1) (NSCLC) and 0.5 mg m(-2) day(-1) (breast cancer) for 21 days every 4 weeks. Three patients achieved a partial response as defined by WHO criteria: one with NSCLC (8%; 95% CI 0-39%) and two with advanced breast cancer (15%; 95% CI 2-45%). The major toxicities were neutropenia and thrombocytopenia, with one episode of neutropenic sepsis. These data suggest that topotecan delivered as a continuous intravenous infusion over 21 days as single-agent therapy does not appear to offer a clinical advantage over conventional 5-day schedules against advanced NSCLC and advanced breast cancer. PMID- 9413955 TI - Discordance between physicians' estimations and breast cancer patients' self assessment of side-effects of chemotherapy: an issue for quality of care. AB - Because side-effects of chemotherapy may be more diverse and patients' reactions more individualistic than tends to be acknowledged by clinicians, a survey was carried out among 50 breast cancer outpatients to document self-reported physical symptoms experienced during NCF (mitoxantrone + cyclophosphamide + 5 fluorouracil) adjuvant chemotherapy and to compare them with the clinicians' estimation in medical records. The questionnaire evaluated the prevalence, duration/severity and distress level of 17 symptoms. Symptom prevalence, assessed in 231 cycles, was high even for symptoms that do not usually focus clinicians' attention. Of these, hot flushes, stomach pain and muscular and articular pains lasted 1 week or more for nearly half of the cycles. Hot flushes, vomiting and stomach pain were the most distressing symptoms. The mean number of symptoms per cycle is significantly correlated with the global quality-of-life score. Concordance between patients' self-assessment and clinical reports, measured in 180 cycles, is moderately correct for vomiting and sore mouth and inadequate for the remaining symptoms even for hair loss (notified in 27% of cycles by clinicians vs 80% by patients) and nausea (38% vs 73%). A better understanding by physicians of cancer patients' problems is necessary to improve quality of care. PMID- 9413956 TI - High serum levels of soluble CD44 variant isoform v5 are associated with favourable clinical outcome in ovarian cancer. AB - In 96 ovarian cancer patients, the present study investigates the clinical significance of pretreatment concentrations of soluble CD44 standard (CD44s) and its isoforms v5 and v6 determined in the serum and the ascitic fluid by means of recently developed enzyme-linked immunosorbent assays (ELISAs). Furthermore, CD44 serum concentrations in the ovarian cancer patients were compared with circulating CD44 levels in 50 healthy age-matched female blood donors. Whereas CD44s was found to be higher and CD44v5 to be lower in ovarian cancer patients than healthy control subjects, no statistical difference between the two cohorts was revealed for CD44 isoform v6. In the ascitic fluid samples, variant isoform v5 and v6 were demonstrated at lower concentrations than serum. Multivariate analysis of overall survival demonstrated that a high pretreatment serum level of soluble CD44 isoform v5 is independently associated with favourable clinical outcome in ovarian cancer. When circulating CD44 isoforms were compared with a panel of serum parameters known to be involved in the immunological network, an inverse correlation between serum CD44v5 levels and indicators of cellular immune system activation, such as soluble interleukin 2 receptor, immunostimulatory protein 90K and neopterin, became apparent. PMID- 9413957 TI - Breast cancer: further metabolic-endocrine risk markers? AB - There is evidence that increased oestrogen receptor (ER) expression in normal mammary epithelium may be a risk marker for the development of breast cancer. Insulin-like growth factor 1 (IGF1) is a potent inducer of mitosis and has been shown to synergize with oestrogen in stimulating the growth of human breast cancer in vitro. In these cells oestradiol has been shown to upregulate IGF type 1 receptor (IGFR), and recently a similar effect has been reported in normal human breast tissue xenografts in vivo. It has been postulated that the combined effect of oestradiol and IGF1 may stimulate proliferation in normal mammary epithelium and increase breast cancer risk. The bioavailability of IGF1 to the tissues is modulated by IGF-binding proteins (IGFBPs), and higher circulating levels of IGF1 and lower levels of IGFBP3 have been reported in breast cancer patients. Breast cancer specimens show a positive correlation between ER status and IGF receptor status, and also a negative correlation between ER status and IGFBP3 expression. Finally, ectopic growth hormone expression has been shown in a majority of specimens of normal and malignant breast tissue, and this may contribute to breast cancer risk, possibly by increasing the local level of bioavailable IGF1. Expansion of such findings may provide clinically useful markers of increased risk to breast cancer in women. PMID- 9413958 TI - Pharmacokinetics and effects on plasma retinol concentrations of 13-cis-retinoic acid in melanoma patients. AB - The pharmacokinetics of 13-cis-retinoic acid (13cisRA) and its effects on retinol plasma levels were investigated after the first and the last doses in melanoma patients, who participated in a study run to assess tolerance over a long period of a treatment schedule of 13cisRA associated with recombinant interferon alpha2a (rIFN-alpha2a). Melanoma patients with regional node metastases after radical surgery were randomized to be treated for 3 months with rIFN-alpha2a, 3 x 10(6) IU s.c. every other day, associated with oral 13cisRA at doses of 20 mg day(-1) (five patients) or 40 mg every other day (seven patients). Maximum 13cisRA blood concentrations usually occurred 4 h after drug administration, with average values of 406 and 633 ng ml(-1) (i.e. 1.3 and 2.1 microM) after the 20 and 40 mg dose respectively. The average half-life (t(1/2)) was approximately 30 h. The maximum concentration, the t(1/2) and the area under the concentration-time curves from 0 to 48 h (AUC(0-48)) of 13cisRA did not change after multiple dosing, whereas the AUC(0-48) of its major blood metabolite, 4-oxo-13-cis retinoic acid, increased. Immediately after 13cisRA treatment, retinol plasma levels started to decline and they reached the lowest values (approximately 20% reduction) shortly after the time of maximum 13cisRA concentrations (i.e. 4-12 h after drug intake). Afterwards, values returned to baseline. The amount of retinol reduction in time was correlated with 13cisRA maximum concentrations. PMID- 9413959 TI - Interstitial pneumonia in patients receiving granulocyte colony-stimulating factor during chemotherapy: survey in Japan 1991-96. AB - Twenty cases of interstitial pneumonia secondary to treatment with granulocyte colony-stimulating factor (G-CSF) were reviewed. Their interstitial pneumonia had the following features: (a) it occurred predominantly in patients aged 60 years or older; (b) it was prevalent among patients with haematological malignancies, particularly non-Hodgkin's lymphoma; (c) in all patients G-CSF was given after anti-cancer agents with potential to affect the lungs; (d) at the onset, many patients had symptoms such as dyspnoea and fever; and (e) the leucocyte (neutrophil) count as well as lactate dehydrogenase (LDH) and C-reactive protein (CRP) levels were usually higher than normal at the onset. These findings indicate that, when G-CSF is used in combination with pneumotoxic anti-cancer agents, respiratory function should be monitored before and during treatment. If the leucocyte (or neutrophil) count and/or LDH and CRP increase suddenly in association with dyspnoea and fever during administration of G-CSF, interstitial pneumonia should be suspected. Accordingly, a chest radiograph and pulmonary functional tests should be performed promptly. If a diagnosis of interstitial pneumonia is made, steroid pulse therapy should be commenced immediately. PMID- 9413961 TI - Interventional neuroradiology: an emerging subspeciality. PMID- 9413960 TI - Preliminary results of the use of intraperitoneal carbon-adsorbed mitomycin C in intra-abdominal malignancy. AB - Eleven patients suffering from intra-abdominal malignancy were treated with various doses of intraperitoneal mitomycin C adsorbed onto activated carbon particles. Seven of the patients underwent resection of their primary gastric tumour and all developed potentially life-threatening severe complications that proved to be fatal in four patients. The pattern of complications seen in these patients was unusual in patients undergoing gastrectomy and must be presumed to be secondary to the intraperitoneal mitomycin C. Intraperitoneal mitomycin C at a dose of 25 mg and 50 mg in the presence of an anastomosis or other suture line does not appear to be safe. PMID- 9413962 TI - Hepatic arterial resistance index--an indicator of diffuse liver disease in children treated with bone marrow transplantation. AB - AIM: To describe the relationship between the resistance index of the common hepatic artery and liver function tests in children undergoing bone marrow transplantation. MATERIALS AND METHODS: We analysed prospectively the results of 106 Doppler ultrasound examinations of the common hepatic artery from 31 bone marrow transplant patients, 16 of whom had normal liver function. The aetiology of the liver dysfunction in the other 15 patients was veno-occlusive disease (n = 7), unknown (n = 3), hepatic graft-versus-host disease (n = 2), hepatitis (n = 2), or cholestasis (n = 1). We assessed the relationships between the hepatic arterial resistance index (HART) and the results of the serum glutamic oxaloacetic transaminase (SGOT), serum glutamic-pyruvic transaminase (SGPT), alkaline phosphatase (ALK), lactate dehydrogenase (LDH) and serum albumin (ALB) assays and calculated HARI break-point values that might distinguish patients with liver disease from patients with the normal liver function. RESULTS: The significant break point (P < 0.05) of the HARI was 0.55 for SGOT, SGPT and ALK. LDH was associated with a break point of 0.53. Resistance indices for the common hepatic artery below the break-point values predicted liver dysfunction with specificities of 81%, 80%, 92% and 93%, respectively. There was no significant relationship between liver function tests and ALB levels. CONCLUSION: If below 0.55, the hepatic arterial resistance index is a non-invasive indicator of liver dysfunction in children undergoing bone marrow transplantation. PMID- 9413963 TI - Safety of percutaneous biopsy of hepatocellular carcinoma with an 18 gauge automated needle. AB - OBJECTIVE: Accurate histological diagnosis and subtyping of hepatocellular carcinoma (hepatoma) is likely to be enhanced if a large biopsy tissue specimen is made available to the pathologist. However biopsy of this tumour can be dangerous, especially if the liver is cirrhotic and the lesion is superficial. This study evaluates the safety of an 18 gauge spring loaded side-cutting needle in the percutaneous biopsy of hepatoma in cirrhotic patients under ultrasonographic (US) guidance. Particular attention was paid to establishing the necessary length of needle track through interposing liver parenchyma to be certain of maximum safety. MATERIALS AND METHODS: One hundred and thirty-nine consecutive biopsy procedures were performed on 129 hepatomas which belonged to 113 men and 12 women of average age 57 +/- 15 years old (median 60, range 8 months-88 years). Ninety-six (69.1%) of these biopsies were performed in cirrhotic livers. The length of biopsy needle track traversing interposing liver parenchyma was less than 1 cm in two cases, 1 cm in 41 cases, between 1 and 2 cm in 46 cases and > 2 cm in 50 cases. The mean tumour size was 7.2 +/- 4.5 cm (median 6.8 cm, range 0.7-25 cm). The average number of needle pass in each biopsy was 2.1 +/- 0.8 times (median 2, range 1-5). RESULTS: One hundred and twenty-six (90.6%) of the biopsy procedures were diagnostic of hepatoma. There were two cases of post-biopsy bleeding, both occurred in procedures with an interposing liver parenchymal track less than 1 cm in length. CONCLUSION: The biopsy technique described was found to be safe for diagnosing hepatoma in patients with or without liver cirrhosis provided that the length of interposing liver parenchymal track is not < 1 cm. PMID- 9413964 TI - Pyogenic liver abscess: treatment with needle aspiration. AB - OBJECTIVE: Percutaneous needle aspiration as an alternative to continuous percutaneous catheter drainage in combination with systemic antibiotics for the treatment of pyogenic liver abscess has never been popular. The authors report their experience with needle aspiration and evaluate its safety, effectiveness and role in treating pyogenic liver abscess. MATERIALS AND METHODS: The results of needle aspiration performed in 101 liver abscesses of 64 unselected consecutive patients with male to female ratio of 2.5:1 and average age 56.3 +/- 16 years were reviewed. The abscesses were pyogenic in 63 patients (98.4%), and multiple in 18 patients (28.1%). Thirty-nine abscesses (38.6%) were > or = 5 cm in diameter. Complete pus removal from each abscess was attempted with 18 gauge thin-walled trocar needles and ultrasound guidance. RESULTS: The percentage of abscesses requiring one, two and three, or more sessions of aspiration was 49.5%, 23.7% and 26.7%, respectively. The overall success rate was 96.8%. The success rate was unrelated to the largest size or number of abscess in the patient. Two patients died from uncontrolled sepsis. One serious complication of liver laceration requiring laparotomy occurred. CONCLUSION: Percutaneous needle aspiration in combination with systemic antibiotics is safe and effective in treating pyogenic liver abscess, it should be considered as a first line alternative to catheter drainage, especially for multiple abscesses. The need for repeat aspirations follows a 'fifty per cent rule'. PMID- 9413965 TI - Colour Doppler sonography of breast masses: a multiparameter analysis. AB - OBJECTIVE: To evaluate signals from breast lesions on both colour and spectral Doppler US, and to correlate findings with angiographic and histopathological features including microvessel density. MATERIALS AND METHODS: One hundred and sixteen breast lesions in 113 patients were evaluated with colour Doppler ultrasonography. Subjective and semi-quantitative assessment of colour signals as well as spectral Doppler analysis were performed and compared. Comparison of colour Doppler features with angiographic and histopathological findings were also carried out. RESULTS: Subjective evaluation revealed that colour signals were more commonly found in malignant (89%) than benign (56%) lesions. Malignant lesions demonstrated a more elaborate vascular network (51%) compared to 12.5% in benign lesions. Spectral Doppler analysis revealed slightly higher specificity values for the pulsatility and resistivity indices, respectively, but lower sensitivity and accuracy values compared to qualitative assessment. Colour Doppler patterns corresponded well with angiograhic images, however, the correlation between colour Doppler parameters and microvessel density was not significant. CONCLUSION: Despite the increased examination time required, spectral analysis does not appear to contribute significantly to the differentiation between malignant and benign breast tumours. Features such as the density, pattern and predominant site of colour signals may be more useful for the evaluation of breast lesions on colour Doppler imaging. PMID- 9413966 TI - Pre-operative localization of breast microcalcification using high-frequency ultrasound. AB - AIM: To determine whether high-frequency ultrasound (US) can be used to reliably localize breast microcalcification preoperatively in the absence of any associated mass lesion or distortion. PATIENTS AND METHODS: Seventeen patients, found to have microcalcification at mammography, the majority screen detected, were studied using high-frequency (10 or 13 MHz) US to visualize the calcified particles. The approximate site of the microcalcifications was first determined from the mammograms. Once the calcifications had been localized, the skin overlying the site was marked with an indelible marker and the depth of the calcifications recorded for the surgeon. Successful excision of microcalcifications was confirmed with postbiopsy specimen X-ray. RESULTS: Fifteen of the 17 patients (88%) underwent successful removal of the microcalcifications. The mean microcalcification cluster size was 160 mm with a mean number of calcifications of 20 at a mean depth of 15 mm. The mean size of the individual calcifications was 0.29 mm. CONCLUSION: Our results show that high frequency US is an effective non-invasive method of identifying and localizing breast microcalcification, and can be used as an alternative to hook wire stereotaxic localization in the majority of patients. PMID- 9413967 TI - Differentiation between benign and metastatic cervical lymph nodes with ultrasound. AB - OBJECTIVE: The differentiation between benign and metastatic lymph nodes with ultrasound (US) is based primarily on the evaluation of size, shape, margin and internal echo structure. The aim of this study is to determine whether these parameters are reliable indicators and to correlate internal echo structure and histopathological findings. MATERIALS AND METHODS: Seventy-one nodes in 21 patients with pathologically proven oral squamous cell carcinoma were examined. The shortest diameter, the short/long diameter ratio (S/L ratio), margins and internal echo structure of the lymph node were evaluated by US. The internal echo structure was divided into six patterns: homogeneous hypoechoic, homogeneous hyperechoic, heterogeneous, eccentric hyperechoic, centric hyperechoic and anechoic pattern. In addition, internal echo structure was correlated with histopathological findings. RESULTS: In 71.4% of the metastatic nodes, the shortest diameter was more than 10 mm and the S/L ratio was higher than that of benign nodes (average 0.71). Eleven (84.6%) of the 13 lymph nodes with irregular margins were metastatic. Heterogeneous and anechoic patterns were observed in metastatic nodes, whereas homogeneous hypoechoic and eccentric hyperechoic patterns were present in benign nodes. On ultrasonography with the corresponding histopathological findings, echogenic areas in the homogeneous hyperechoic, heterogeneous and centric hyperechoic patterns of metastatic nodes proved to be necrosis or fibrosis. Eccentric hyperechoic areas in benign nodes corresponded to the hilus and surrounding fatty tissue. CONCLUSIONS: The shortest diameter, S/L ratio, margin and internal echo structure were considered to be critical indicators to differentiate between benign and metastatic nodes. Secondary changes caused by tumour infiltration, necrosis, or fibrosis should be assessed when metastatic lymph nodes are differentiated from benign ones by internal echo structure. PMID- 9413968 TI - Local staging of prostate cancer with 0.2 T body coil MRI. AB - This study was undertaken to evaluate the accuracy of low field strength body coil MRI in the staging of clinically localized prostate cancer. Fifty-three patients with prostate cancer were examined on a 0.2 T body coil system before undergoing radical prostatectomy. Of the 20 cases with unconfined stage T3 disease on histology, 12 were correctly staged, whilst three cases were overstaged by MRI. (Accuracy 79.2%, sensitivity 60%, and specificity 90.9%.) The accuracy, sensitivity and specificity for the detection of capsular penetration were 77.3%, 55% and 90.9%, respectively, whilst those for seminal vesical invasion were 94.3%, 83.3% and 95.7%, respectively. It is concluded that a high level of staging accuracy, comparable to that obtained in some published studies using high field strength endorectal coil MRI, can be obtained using 0.2 T body coil MRI. PMID- 9413969 TI - Percutaneous laser discectomy: MR findings within the first 24 hours after treatment and their relationship to clinical outcome. AB - OBJECTIVE: The purpose of this study was to evaluate the early MR appearances of the intervertebral disc obtained within 24 h after percutaneous laser discectomy and to determine if a correlation exists between the MR images and the clinical outcome. SUBJECTS AND METHODS: Twenty-nine discs in 26 patients with contained lumbar disc herniation treated by laser were included. Laser intervention was performed using Ho:YAG laser system. The results were quantitatively analysed by measuring areal changes of the herniated mass on axial T1-weighted images and signal changes within the disc on sagittal T2-weighted images. Clinical outcomes were evaluated according to Japanese Orthopaedic Association score (JOA score), and recovery rates based on JOA score immediately and 1 year after treatment were calculated. RESULTS: The recovery rate immediately after treatment was 53.1%, and increasing to 64.6% 1 year later. The size of the disc herniation ranged from 7% to 55% of the axial cross-section of the spinal canal pre-operatively and no significant changes were noted postoperatively. Also no correlation was found between the pre-operative herniation size and the recovery rate. The signal changes within the disc increased significantly after treatment, but no correlation was present between the signal changes and the recovery rate. In five patients, soft-tissue changes along the laser tract were well demonstrated on MR imaging. CONCLUSION: Although immediate postoperative MR imaging shows early tissue changes induced by laser, our study has not proven that immediate postoperative MR imaging could predict the clinical outcome after percutaneous laser discectomy. PMID- 9413970 TI - The use of play therapy in the preparation of children undergoing MR imaging. AB - Magnetic resonance (MR) imaging has become an important technique in the evaluation of a wide range of congenital and acquired conditions in children. The ability to image in multiple anatomic planes without the use of ionizing radiation has particular advantages. However scan times can be long, and the narrow bore and noise generated by most units make the procedure very intimidating to the young child. The use of surface coils may exacerbate this, often necessitating recourse to sedation or anaesthesia. We describe a technique involving play therapy which we have found useful in the preparation of young children for MR imaging and which has reduced the number of non-diagnostic scans and the need for sedation or anaesthesia. PMID- 9413971 TI - Gadopentate dimeglumine as an alternative contrast agent for use in interventional procedures. AB - Renal failure due to iodinated contrast media is a serious complication of peripheral angiography in patients with impaired renal function. Gadolinium based agents which are widely used in magnetic resonance imaging have no adverse renal toxicity at recommended doses. These agents have sufficient radiographic density to be seen using digital subtraction equipment. We describe the use of undiluted gadopentate dimeglumine as the radiographic contrast agent in two patients who underwent complex interventional procedures. PMID- 9413972 TI - Suprarenal cystic masses: unusual causes. AB - Two cases of large suprarenal cystic masses with extremely rare histological diagnoses of adrenal neurogenous choristoma and multicystic intrarenal neuroblastoma are presented. Ultrasonographic, CT scanning and histological features are described and differential diagnosis discussed. PMID- 9413973 TI - Case report: CT demonstration of supra-diaphragmatic calcified metastatic nodes from ovarian carcinoma. PMID- 9413974 TI - Case report: giant dorsal neurofibroma in a young male. PMID- 9413975 TI - Case report: nodular fasciitis of the breast. PMID- 9413976 TI - Case report: gallbladder carcinoma--an unusual cause of haemobilia. PMID- 9413977 TI - Manual compression of femoral pseudoaneurysms. PMID- 9413978 TI - Use of gadolinium enhancement MRI in postoperative lumbar spine assessment. PMID- 9413979 TI - Phytochrome: if it looks and smells like a histidine kinase, is it a histidine kinase? PMID- 9413980 TI - Putting it all together: building a prereplicative complex. PMID- 9413981 TI - How do viruses enter cells? The HIV coreceptors teach us a lesson of complexity. PMID- 9413982 TI - Molecular mechanisms of endocytosis. PMID- 9413983 TI - The yeast splice site revisited: new exon consensus from genomic analysis. PMID- 9413984 TI - Solution structure of the KIX domain of CBP bound to the transactivation domain of CREB: a model for activator:coactivator interactions. AB - The nuclear factor CREB activates transcription of target genes in part through direct interactions with the KIX domain of the coactivator CBP in a phosphorylation-dependent manner. The solution structure of the complex formed by the phosphorylated kinase-inducible domain (pKID) of CREB with KIX reveals that pKID undergoes a coil-->helix folding transition upon binding to KIX, forming two alpha helices. The amphipathic helix alphaB of pKID interacts with a hydrophobic groove defined by helices alpha1 and alpha3 of KIX. The other pKID helix, alphaA, contacts a different face of the alpha3 helix. The phosphate group of the critical phosphoserine residue of pKID forms a hydrogen bond to the side chain of Tyr-658 of KIX. The structure provides a model for interactions between other transactivation domains and their targets. PMID- 9413985 TI - Ectopically expressed CAG repeats cause intranuclear inclusions and a progressive late onset neurological phenotype in the mouse. AB - The mutations responsible for several human neurodegenerative disorders are expansions of translated CAG repeats beyond a normal size range. To address the role of repeat context, we have introduced a 146-unit CAG repeat into the mouse hypoxanthine phosphoribosyltransferase gene (Hprt). Mutant mice express a form of the HPRT protein that contains a long polyglutamine repeat. These mice develop a phenotype similar to the human translated CAG repeat disorders. Repeat containing mice show a late onset neurological phenotype that progresses to premature death. Neuronal intranuclear inclusions are present in affected mice. Our results show that CAG repeats do not need to be located within one of the classic repeat disorder genes to have a neurotoxic effect. PMID- 9413986 TI - Oral somatic transgene vaccination using attenuated S. typhimurium. AB - An attenuated strain of S. typhimurium has been used as a vehicle for oral genetic immunization. Eukaryotic expression vectors containing truncated genes of ActA and listeriolysin--two virulence factors of Listeria monocytogenes--have been used to transform S. typhimurium aroA. Multiple or even single oral immunizations with such transformants induced excellent cellular and humoral responses. In addition, protective immunity was induced with listeriolysin transformants. The quality of the responses suggested a transfer of plasmid DNA from the bacterial carrier to the host. Such transfer was unequivocally shown in vitro with primary peritoneal macrophages. We describe a highly versatile system for antigen delivery, identification of protective antigens for vaccination, and efficient generation of antibodies against the product of open reading frames present on virtually any DNA segment. PMID- 9413987 TI - Drosophila ecdysone receptor mutations reveal functional differences among receptor isoforms. AB - The steroid hormone ecdysone directs Drosophila metamorphosis via three heterodimeric receptors that differ according to which of three ecdysone receptor isoforms encoded by the EcR gene (EcR-A, EcR-B1, or EcR-B2) is activated by the orphan nuclear receptor USP. We have identified and molecularly mapped two classes of EcR mutations: those specific to EcR-B1 that uncouple metamorphosis, and embryonic-lethal mutations that map to common sequences encoding the DNA- and ligand-binding domains. In the larval salivary gland, loss of EcR-B1 results in loss of activation of ecdysone-induced genes. Comparable transgenic expression of EcR-B1, EcR-B2, and EcR-A in these mutant glands results, respectively, in full, partial, and no repair of that loss. PMID- 9413988 TI - Exocrine gland dysfunction in MC5-R-deficient mice: evidence for coordinated regulation of exocrine gland function by melanocortin peptides. AB - The effects of pituitary-derived melanocortin peptides are primarily attributed to ACTH-mediated adrenocortical glucocorticoid production. Identification of a widely distributed receptor for ACTH/MSH peptides, the melanocortin-5 receptor (MC5-R), suggested non-steroidally mediated systemic effects of these peptides. Targeted disruption of the MC5-R produced mice with a severe defect in water repulsion and thermoregulation due to decreased production of sebaceous lipids. High levels of MC5-R was found in multiple exocrine tissues, including Harderian, preputial, lacrimal, and sebaceous glands, and was also shown to be required for production and stress-regulated synthesis of porphyrins by the Harderian gland and ACTH/MSH-regulated protein secretion by the lacrimal gland. These data show a requirement for the MC5-R in multiple exocrine glands for the production of numerous products, indicative of a coordinated system for regulation of exocrine gland function by melanocortin peptides. PMID- 9413989 TI - Molecular basis for the binding promiscuity of an anti-p24 (HIV-1) monoclonal antibody. AB - Multiple binding capabilities utilized by specific protein-to-protein interactions in molecular recognition events are being documented increasingly but remain poorly understood at the molecular level. We identified five unrelated peptides that compete with each other for binding to the paratope region of the monoclonal anti-p24 (HIV-1) antibody CB4-1 by using a synthetic positional scanning combinatorial library XXXX[B1,B2,B3,X1,X2,X3]XXXX (14 mers; 68,590 peptide mixtures in total) prepared by spot synthesis. Complete sets of substitution analogs of the five peptides revealed key interacting residues, information that led to the construction of binding supertopes derived from each peptide. These supertope sequences were identified in hundreds of heterologous proteins, and those proteins that could be obtained were shown to bind CB4-1. Implications of these findings for immune escape mechanisms and autoimmunity are discussed. PMID- 9413990 TI - Crystallographic analysis of anti-p24 (HIV-1) monoclonal antibody cross reactivity and polyspecificity. AB - The X-ray crystal structures of an anti-p24 (HIV-1) monoclonal antibody Fab fragment alone and in complexes with the epitope peptide GATPQDLNTnL (n = norleucine), an epitope-homologous peptide GATPEDLNQKLAGN, as well as two unrelated peptides GLYEWGGARITNTD and efslkGpllqwrsG (D-peptide), are presented to a maximum resolution of 2.6 A. The latter three peptides were identified from screening synthetic combinatorial peptide libraries. Although all peptides bind to the same antigen combining site, the nonhomologous peptides adopt different binding conformations and also form their critical contacts with different antibody residues. Only small readjustments are observed within the framework of the Fab fragment upon binding. PMID- 9413991 TI - Novel disease resistance specificities result from sequence exchange between tandemly repeated genes at the Cf-4/9 locus of tomato. AB - Tomato Cf genes confer resistance to C. fulvum, reside in complex loci carrying multiple genes, and encode predicted membrane-bound proteins with extracytoplasmic leucine-rich repeats. At least two Cf-9 homologs confer novel C. fulvum resistance specificities. Comparison of 11 genes revealed 7 hypervariable amino acid positions in a motif of the leucine-rich repeats predicted to form a beta-strand/beta-turn in which the hypervariable residues are solvent exposed and potentially contribute to recognition specificity. Higher nonsynonymous than synonymous substitution rates in this region imply selection for sequence diversification. We propose that the level of polymorphism between intergenic regions determines the frequency of sequence exchange between the tandemly repeated genes. This permits sufficient exchange to generate sequence diversity but prevents sequence homogenization. PMID- 9413992 TI - Disruption of alpha3 connexin gene leads to proteolysis and cataractogenesis in mice. AB - Gap junction channels formed by alpha3 (Cx46) and alpha8 (Cx50) connexin provide pathways for communication between the fiber cells in the normal transparent lens. To determine the specific role of alpha3 connexin in vivo, the alpha3 connexin gene was disrupted in mice. Although the absence of alpha3 connexin had no obvious influence on the early stages of lens formation and the differentiation of lens fibers, mice homozygous for the disrupted alpha3 gene developed nuclear cataracts that were associated with the proteolysis of crystallins. This study establishes the importance of gap junctions in maintaining normal lens transparency by providing a cell-cell signaling pathway or structural component for the proper organization of lens membrane and cytoplasmic proteins. PMID- 9413993 TI - Association of transcriptionally silent genes with Ikaros complexes at centromeric heterochromatin. AB - Ikaros proteins are required for normal T, B, and NK cell development and are postulated to activate lymphocyte-specific gene expression. Here we examined Ikaros distribution in the nucleus of B lymphocytes using confocal microscopy and a novel immunofluorescence in situ hybridization (immuno-FISH) approach. Unexpectedly, Ikaros localized to discrete heterochromatin-containing foci in interphase nuclei, which comprise clusters of centromeric DNA as defined by gamma satellite sequences and the abundance of heterochromatin protein-1 (HP-1). Using locus-specific probes for CD2, CD4, CD8alpha, CD19, CD45, and lambda5 genes, we show that transcriptionally inactive but not transcriptionally active genes associate with Ikaros-heterochromatin foci. These findings support a model of organization of the nucleus in which repressed genes are selectively recruited into centromeric domains. PMID- 9413994 TI - Cellular construction of a circadian clock: period determination in the suprachiasmatic nuclei. AB - The circadian clock in the suprachiasmatic nuclei is composed of multiple, single cell circadian oscillators (clock cells). We now test the hypothesis that the circadian period in behavior is determined by the mean period that arises from the coupling of clock cells with diverse circadian periods. For these studies, we monitored firing rate rhythms of individual suprachiasmatic nuclei neurons on fixed multielectrode plates and exploited the altered circadian periods expressed by heterozygous and homozygous tau mutant hamsters. The results show that circadian period in the whole animal is determined by averaging widely dispersed periods of individual clock cells. The data also demonstrate that the tau mutation affects circadian function in a cell-autonomous manner. PMID- 9413995 TI - Phage DNA packaging. AB - Phage DNA packaging occurs by DNA translocation into a preformed protein shell--a prohead--with the aid of a packaging enzyme or a terminase. The packaging enzyme is composed of two subunits: the large subunit has ATP-binding, prohead binding, and DNA cleavage activities, and the small subunit is a DNA binding protein. DNA translocation is driven by ATP hydrolysis. In general, phage DNA replication mechanisms lead to the accumulation of concatemers. Concatemers are processed to mature DNA during and depending upon DNA packaging. This review will focus on the molecular mechanism of concatemer processing and the coupling of ATP hydrolysis to DNA translocation. PMID- 9413996 TI - Inhibition of transpositional recombination by OrfA and OrfB proteins encoded by insertion sequence IS3. AB - BACKGROUND: An insertion element IS3 is flanked by terminal inverted repeat (IR) sequences. IS3 encodes two, out-of-phase, overlapping open reading frames, orfA and orfB, from which three proteins are produced. OrfAB is a transframe protein produced by -1 translational frameshifting between orfA and orfB, and it is known to be IS3 transposase. OrfA and OrfB are the proteins produced without frameshifting, but their functions have not been elucidated. RESULTS: A plasmid carrying an IS3 mutant that produces only transposase generates miniplasmids- which are the IS3-mediated intramolecular transposition products--as well as characteristic IS3 circles and linear IS3 molecules. OrfA inhibited the generation of these small molecules to a lesser degree, but OrfB did not. OrfB, together with OrfA, however, inhibited the generation more strongly than OrfA alone. OrfA also inhibited the intermolecular transposition of mini-IS3 with the chloramphenicol-resistance gene flanked by IRs to a reduced frequency, and OrfB together with OrfA inhibited it almost completely. OrfA and/or OrfB did not, however, repress transcription from the promoter in the left-terminal region preceding orfA. CONCLUSIONS: The results obtained above show that OrfA and OrfB are not repressors but are inhibitors of transpositional recombination promoted by transposase. OrfA with an alpha helix-turn-alpha helix DNA-binding motif may compete with transposase to bind to terminal IRs. OrfA, together with OrfB that has a DDE motif conserved in retroviral integrases, may inhibit the formation of an active transpososome consisting oftransposase, two terminal IRs and target DNA for the strand transfer reaction. IS3 with a limited size, 1258 bp in length, uses strategies of translational frameshifting and coupling to produce transposase as well as negative regulators to make its copies at a low level, which minimizes a deleterious effect of transposition on bacterial hosts. PMID- 9413997 TI - Temporal regulation of the mid-prepupal gene FTZ-F1: DHR3 early late gene product is one of the plural positive regulators. AB - BACKGROUND: Various ecdysteroid responsive genes play important roles in insect moulting and metamorphosis. Late FTZ-F1, a member of the nuclear receptor superfamily, is a unique transcription factor which is induced by a pulse exposure of 20-hydroxyecdysone. Elucidation of the regulation mechanism of this gene during prepupal period will help our understanding of metamorphosis at a molecular level. RESULTS: Using transgenic fly lines carrying various transcription regulatory regions of the FTZ-F1 gene fused to the LacZ gene, we investigated cis-regulatory elements in the late FTZ-F1 transcription unit. The region which governs the stage-specific expression during prepupal period was narrowed down to 1.2kb, from -0.7 to +0.5kb relative to the transcription start site. Electrophoresis mobility shift assays using staged extracts and various probes within the stage-specific region allowed us to identify binding sites for DHR3, an early late gene product, around 170 and 450bp downstream of the transcription initiation site. Mutations disrupting these binding sites reduced the reporter gene expression without affecting the stage specificity. CONCLUSIONS: Our deletion and mutation studies of the cis-regulatory element of the FTZ-F1 gene suggest that the DHR3 binding sites located in the 5' non-coding region are involved in the prepupal expression of the gene. These DHR3 binding sites confer high level expression while other elements are also involved in stage-specific expression. PMID- 9413998 TI - Characterization of the mouse prostaglandin F receptor gene: a transgenic mouse study of a regulatory region that controls its expression in the stomach and kidney but not in the ovary. AB - BACKGROUND: The actions of prostaglandin F2alpha are mediated by a cell-surface receptor (FP), but little is known about the regulation of FP gene expression. To clarify the mechanisms underlying tissue specific transcription of the mouse FP gene, we isolated and characterized mouse genomic DNA clones encoding FP. RESULTS: Structural analysis revealed that the mouse FP gene is composed of three exons and two introns, and spans more than 11 kilobases. By primer extension and PCR analyses, the major transcription start site was identified as a cytosine nucleotide, but additional sites of transcription initiation were found in the ovary. There was no apparent difference in the FP gene transcription initiation site between the ovary, kidney and stomach. Sequence analysis of the putative promoter region showed only two potential SP-1 binding sites, but no other typical well-known consensus sequences. We generated transgenic mice with the potential promoter region of the FP gene connected to the lacZ reporter gene. Northern blot analysis showed that the pattern of expression of the transgene corresponded to that of FP expression, except in the ovary. Upon analysis by in situ hybridization, the lacZ gene transcripts were found to be expressed in the fundic glands in the stomach, and the cortical tubules in the kidney, in which endogenous FP transcripts were also expressed. On the contrary, expression of lacZ transcripts was not detected in the corpora lutea, where the highest expression of FP mRNA was observed. CONCLUSIONS: These studies suggest that a separate control mechanism exists for FP expression in the ovary, distinct from the expression in the stomach and kidney. PMID- 9413999 TI - A novel form of the myeloid-specific zinc finger protein (MZF-2). AB - BACKGROUND: Myeloid cell development is controlled by tissue-specific transcription factors. Human myeloid zinc finger protein (MZF-1) is a putative transcription factor containing 13 zinc fingers, and has been suggested that it regulates the development of neutrophilic granulocytes. RESULTS: Here, we have isolated the murine and human cDNAs which encode a novel form of MZF protein (MZF 2). Murine and human MZF-2 proteins consisted of 814 and 775 amino acids, respectively, and have identity of 75.3% between them. The C-terminal half of human MZF-2, carrying the zinc finger domains, was completely identical with that of human MZF-1, whereas the N-terminal half of MZF-2 was different from the corresponding region of human MZF-1, and was coded by distinct exons. MZF-2 mRNA was expressed in myeloid cells, particularly in the cells committed to the neutrophilic lineage, and down-regulated by G-CSF. CONCLUSIONS: MZF-1 and MZF-2 mRNAs seem to be produced by the alternative use of two different transcription initiation sites. The distinct N-terminal half of MZF-2 carries two characteristic domains, a leucine-rich domain called LeR and an acidic domain, which suggests a unique function of MZF-2 in neutrophil development. PMID- 9414000 TI - Divergent cortical connections to entorhinal cortex from area TF in the macaque. AB - The entorhinal cortex (EC) is an important component of the medial temporal lobe memory system in the primate and is often viewed as a "gatekeeper" area that passes on highly convergent cortical inputs toward the hippocampus. Further analysis of these connections at a microcircuitry level regarding the actual size and shape of arbors and terminations is not yet available, but may contribute to understanding the role of the EC in memory or other functions. The main emphasis of this report was on serial section analysis of anterogradely labeled axons that project from area TF (lateral parahippocampal cortex; Bonin and Bailey, 1947) to the EC (n = 12). By way of evaluating network organization, other projections from area TF--to TH (in the medial parahippocampal gyrus; n = 5) and to posterior visual areas (n = 3)--were also investigated. All three systems were found to terminate heavily in layer 1, as expected from previous investigations, but some terminations were verified in layer 6 of the EC as well. This technique further demonstrated that terminal fields are widely divergent and elongated. In the EC, terminal fields extended over 6-11 mm and spanned multiple cell islands and interislands. These axons resemble "feedback" cortical connections by virtue of their layer 1 terminations and their markedly divergent geometry, but not by their origin from layer 3. Spatially extended terminal fields recall the nontopographic, distributed character of olfactory connections and raise questions of how these features might be related to the memory functions attributed to medial temporal regions. PMID- 9414001 TI - Localization of last-order premotor interneurons in the lumbar spinal cord of rats. AB - There is strong evidence that neural circuits underlying certain rhythmic motor behaviors are located in the spinal cord. Such local central pattern generators are thought to coordinate the activity of motoneurons through specific sets of last-order premotor interneurons that establish monosynaptic contacts with motoneurons. After injections of biotinylated dextran amine into the lateral and medial motor columns as well as the ventrolateral white matter at the level of the upper and lower segments of the lumbar spinal cord, we intended to identify and localize retrogradely labelled spinal interneurons that can likely be regarded as last-order premotor interneurons in rats. Regardless of the location of the injection site, labelled interneurons were revealed in laminae V-VIII along a three- or four-segment-long section of the spinal gray matter. Although most of the stained cells were confined to laminae V-VIII in all cases, the distribution of neurons within the confines of this area varied according to the site of injection. After injections into the lateral motor column at the level of the L4-L5 segments, the labelled neurons were located almost exclusively in laminae V-VII ipsilateral to the injection site, and the perikarya were distributed throughout the entire mediolateral extent of this area. Interneurons projecting to the lateral motor column at the level of the L1-L2 segments were also located in laminae V-VII, but most of them were concentrated in the middle one-third or in the lateral half of this area. Following injections into the medial motor column at the level of the L1-L2 segments, the majority of labelled neurons were confined to the medial aspect of laminae V-VII and lamina VIII, and the proportion of neurons that were found contralateral to the injection site was strikingly higher than in the other experimental groups. The results suggest that the organization of last-order premotor interneurons projecting to motoneurons, which are located at different areas of the lateral and medial motor columns and innervate different muscle groups, may present distinct features in the rat spinal cord. PMID- 9414002 TI - Microglia development in the quail cerebellum. AB - We used the QH1 antibody to study changes in the morphological features and distribution of microglial cells throughout development in the quail cerebellum. Few microglial precursors were present in the cerebellar anlage before the ninth incubation day (E9), whereas many precursors apparently entered the cerebellum from the meninges in the basal region of the cerebellar peduncles between E9 and E16. From this point of entry into the nervous parenchyma, they spread through the cerebellar white matter, forming a 'stream' of labeled cells that could be seen until hatching (E16). The number of microglial cells in the cerebellar cortex increased during the last days of embryonic life and first posthatching week, whereas microglial density within the white matter decreased after hatching. As a consequence, the differences in microglial cell density observed in the cerebellar cortex and the white matter during embryonic life diminished after hatching, and microglia showed a nearly homogeneous pattern of distribution in adult cerebella. Ameboid and poorly ramified microglial cells were found in developing stages, whereas only mature microglia appeared in adult cerebella. Our observations suggest that microglial precursors enter the cerebellar anlage mainly by traversing the pial surface at the basal region of the peduncles, then migrate along the white matter, and finally move radially to the different cortical layers. Differentiation occurs after the microglial cells have reached their final position. In other brain regions the development of microglia follows similar stages, suggesting that these steps are general rules of microglial development in the central nervous system. PMID- 9414003 TI - Developmental changes in GABA neurons of the rat dentate gyrus: an in situ hybridization and birthdating study. AB - The temporal and spatial distribution of glutamate decarboxylase 67 (GAD67) mRNA containing neurons in the dentate gyrus was analyzed from embryonic day 20 (E20) to postnatal day 15 (PN15) with nonradioactive in situ hybridization methods. A major goal was to follow the development of an early-appearing population of gamma-aminobutyric acid (GABA) neurons within the developing molecular layer. At E20, GAD67 mRNA-containing neurons were highly concentrated slightly above the outer border of the developing granule cell layer. By PN3-PN5, many labeled cells were distributed within the developing granule cell layer; by PN15, labeled neurons occupied positions normally seen in the adult, such as along the inner border of the granule cell layer. This developmental pattern is unique and led to additional studies to determine the potential fate of the early-appearing GABA population. The possibility of apoptotic cell death was investigated with in situ end labeling techniques at developmental ages E21-PN15. Very few apoptotic cells were detected in the developing molecular layer at all ages examined. Birthdating studies of neurons labeled with bromodeoxyuridine revealed a changing pattern, similar to that of GAD67 mRNA, for cells with birthdates (E14) of many of the mature GAD-containing neurons in the dentate gyrus. The changes in the mRNA and birthdating patterns from E20-PN15 suggest that some of the early-appearing GABA neurons within the developing molecular layer of the dentate gyrus may alter their positions to eventually become the mature GABA population along the inner border of the granule cell layer. PMID- 9414004 TI - A regional ultrastructural analysis of the cellular and synaptic architecture in the chinchilla cristae ampullares. AB - The chinchilla crista ampullaris was studied in 10 samples, each containing 32 consecutive ultrathin sections of the entire neuroepithelium. Dissector methods were used to estimate the incidence of various synaptic features, and results were confirmed in completely reconstructed hair cells. There are large regional variations in cellular and synaptic architecture. Type I and type II hair cells are shorter, broader, and less densely packed in the central zone than in the intermediate and peripheral zones. Complex calyx endings are most common centrally. On average, there are 15-20 ribbon synapses and 25-30 calyceal invaginations in each type I hair cell. Synapses and invaginations are most numerous centrally. Central type II hair cells receive considerably fewer afferent boutons than do peripheral type II hair cells, but have similar numbers of ribbon synapses. The numbers are similar because central type II hair cells make more synapses with the outer faces of calyx endings and with individual afferent boutons. Most afferent boutons get one ribbon synapse. Boutons without ribbon synapses were only found peripherally, and boutons getting multiple synapses were most frequent centrally. Throughout the neuroepithelium, there is an average of three to four efferent boutons on each type II hair cell and calyx ending. Reciprocal synapses are rare. Most synaptic ribbons in type I hair cells are spherules; those in type II hair cells can be spherical or elongated and are particularly heterogeneous centrally. Consistent with the proposal that the crista is concentrically organized, the intermediate and peripheral zones are each similar in their cellular and synaptic architecture near the base and near the planum. An especially differentiated subzone may exist in the middle of the central zone. PMID- 9414005 TI - Distribution and morphology of luteinising hormone-releasing hormone neurones in a species of wild antelope, the springbok (Antidorcas marsupialis). AB - The distribution and morphology of luteinising hormone-releasing hormone (LHRH) neurones varies between species. The primary purpose of this study was to characterize the distribution and morphology of the LHRH system in a species of antelope, the springbok. This wild antelope has a well-defined social structure in which reproductive activity is confined to a few dominant, territorial rams. We also sought to determine whether social or reproductive status could be accounted for by differences in the distribution or morphology of hypothalamic LHRH neurones. Eleven anoestrous female, nine breeding territorial male (TM) and eight "bachelor" male (BM) springbok were obtained, and their reproductive and body conditions were assessed. By using standard immunocytochemical techniques, the LHRH system was visualised in the brains of four animals from each group. Immunoreactive neurones were located in a continuum from the septum to the arcuate nucleus, with the majority at the level of the organum vasculosum of the lamina terminalis. Neither the distribution nor the number of cells differed among the three groups. Furthermore, the area of LHRH perikarya was similar in both groups of males, suggesting that reproductive differences between TMs and BMs lie at another level of the neuroendocrine axis. The anoestrous females had significantly larger neurones than males (TM plus BM). This may reflect a sex difference in the LHRH system of this wild antelope. However, an alternative explanation is that the male/female difference is related to the comparatively inactive reproductive neuroendocrine state of the females. PMID- 9414006 TI - Labeling of pyramidal and nonpyramidal neurons with lectin Vicia villosa during postnatal development of the guinea pig. AB - Labeling of cortical neurons with a lectin, Vicia villosa (VVA), was investigated in guinea pigs aged 1 day old to adult. Lectin histochemistry revealed a perineuronal sheath, which outlined the cell bodies, apical dendrites, and axon initial segments, in distinct populations of pyramidal and nonpyramidal neurons. Their laminar positions were segregated, with the pyramidal neurons confined to layer 5 and the nonpyramidal neurons distributed mainly in layers 3-5. The VVA labeled substance(s) was detected at the interfaces between neurons and cellular elements present in the perisynaptic region, including glial processes and fine axons. However, it was excluded from synaptic clefts of presynaptic terminals. Double immunohistochemistry revealed that most of the VVA-labeled neurons were also labeled perineuronally with a monoclonal antibody, Cat 301, and vice versa. Dendritic patterns of the VVA-labeled pyramidal neurons were studied further by intracellular injection of Lucifer yellow into fixed slices. Apical dendrites had a considerable thickness before arborizing into a few daughter branches in layer 3 or 4, suggesting a morphological resemblance to intrinsic, bursting pyramidal neurons defined physiologically in vitro. During postnatal development, there was a global spatiotemporal pattern in the onset of VVA labeling of the cortical neurons. The labeling progressed from medial and posterior cortical areas, which are closely related to the hippocampal formation, to more lateral and anterior areas, which are less closely related. The labeling patter thus tends to follow the order of the phylogenetical development of the isocortex. PMID- 9414007 TI - Rapid regulation of cytoskeletal proteins and their mRNAs following afferent deprivation in the avian cochlear nucleus. AB - During development, removal of neuronal input can lead to profound changes in postsynaptic cells, including atrophy and cell death. In the chicken brainstem cochlear nucleus, the nucleus magnocellularis (NM), deprivation of auditory input via unilateral cochlea removal or silencing the eighth nerve with tetrodotoxin leads to a loss of 25-30% of the neurons and the atrophy of surviving neurons. One intracellular component that may be involved in both cell atrophy and cell death is the cytoskeleton. The degradation of the cytoskeleton following deafferentation could potentially lead to either atrophy or death of NM neurons. However, little is known regarding the role of neuronal input on the cytoskeletal structure of NM neurons and whether changes in the cytoskeleton are responsible for cell death following deafferentation. The present study examined whether changes in the cytoskeleton of NM neurons occurred following cochlea removal. Several components of the cytoskeleton were analyzed following unilateral afferent deprivation. Levels of immunostaining for tubulin, actin, and microtubule-associated protein 2 (MAP-2), and levels of beta-tubulin and beta actin mRNAs were assessed in NM neurons following cochlea removal. Our results revealed that afferent deprivation results in a rapid decrease in immunostaining for all three cytoskeletal proteins examined. These decreases were observed as early as 3 hours after cochlea removal and persisted for up to 4 days. In addition, these changes occurred in all deafferented NM neurons at the early time points, indicating that both dying and surviving NM neurons undergo a similar change in their cytoskeletons. In contrast to the decreases in immunostaining, levels of beta-tubulin and beta-actin mRNAs were not noticeably altered by deafferentation. Our findings indicate that the cytoskeleton is altered or degraded following deafferentation but that this process is not regulated at the transcriptional level. PMID- 9414008 TI - Parasympathetic innervation of cephalic arteries in rabbits: comparison with sympathetic and sensory innervation. AB - We investigated the distribution of parasympathetic, sympathetic, and sensory perivascular nerve fibers in rabbit cephalic arteries supplying the brain, exocrine glands, nasal mucosa, masseter muscles, tongue, and skin in the face and also examined cranial autonomic and sensory ganglia. NADPH diaphorase (NADPHd) positive and vasoactive intestinal peptide-like immunoreactive (VIP-LI) neurons were located in the cranial parasympathetic ganglia. Neuropeptide Y (NPY)-LI neurons occurred mainly, and dopamine beta-hydroxylase (DBH)-LI neurons occurred exclusively, in the superior cervical (sympathetic) ganglion. Substance P (SP)-LI and calcitonin gene-related peptide (CGRP)-LI neurons occurred only in the trigeminal (sensory) ganglion. Therefore, it was assumed that NADPHd-positive and VIP-LI perivascular nerve fibers in cephalic arteries were parasympathetic, all DBH-LI and most NPY-LI fibers were sympathetic, and SP-LI and CGRP-LI fibers were sensory in nature. In the cerebral arteries, NADPHd-positive and VIP-LI varicose fibers were more numerous in the rostral than in the caudal half of the Circle of Willis. In the extracranial arteries, NADPHd-positive and VIP-LI fibers were most abundant in the lingual, lacrimal, and supraorbital arteries; sparse in the parotid and submandibular arteries; and absent in the ear artery. There was an obvious proximal-to-distal density gradient along individual cephalic arterial trees. In contrast, DBH-LI, NPY-LI, SP-LI, and CGRP-LI varicose nerve fibers were similar in density in all cephalic arteries and their branches. These neuroanatomical findings suggest that differential parasympathetic innervation in cephalic arteries may play a role in the partitioning of blood flow between different cephalic tissues. PMID- 9414010 TI - Organization of neural inputs to the suprachiasmatic nucleus in the rat. AB - The circadian timing of the suprachiasmatic nucleus (SCN) is modulated by its neural inputs. In the present study, we examine the organization of the neural inputs to the rat SCN using both retrograde and anterograde tracing methods. After Fluoro-Gold injections into the SCN, retrogradely labeled neurons are present in a number of brain areas, including the infralimbic cortex, the lateral septum, the medial preoptic area, the subfornical organ, the paraventricular thalamus, the subparaventricular zone, the ventromedial hypothalamic nucleus, the posterior hypothalamic area, the intergeniculate leaflet, the olivary pretectal nucleus, the ventral subiculum, and the median raphe nuclei. In the anterograde tracing experiments, we observe three patterns of afferent termination within the SCN that correspond to the photic/raphe, limbic/hypothalamic, and thalamic inputs. The median raphe projection to the SCN terminates densely within the ventral subdivision and sparsely within the dorsal subdivision. Similarly, areas that receive photic input, such as the retina, the intergeniculate leaflet, and the pretectal area, densely innervate the ventral SCN but provide only minor innervation of the dorsal SCN. A complementary pattern of axonal labeling, with labeled fibers concentrated in the dorsal SCN, is observed after anterograde tracer injections into the hypothalamus and into limbic areas, such as the ventral subiculum and infralimbic cortex. A third, less common pattern of labeling, exemplified by the paraventricular thalamic afferents, consists of diffuse axonal labeling throughout the SCN. Our results show that the SCN afferent connections are topographically organized. These hodological differences may reflect a functional heterogeneity within the SCN. PMID- 9414009 TI - Neurokinin 1 receptor expression in the rat retina. AB - Tachykinin (TK) peptides influence neuronal activity in the inner retina of mammals. The aim of this investigation was to determine the cellular localization of the neurokinin 1 receptor (NK1), whose preferred ligand is the TK peptide substance P (SP), in the rat retina. These studies used a polyclonal antiserum directed to the C-terminus of rat NK1. The majority of NK1-immunoreactive (IR) cells were located in the proximal inner nuclear layer (INL), and very rarely they were found in the distal INL. Some small and large NK1-IR somata were present in the ganglion cell layer. NK1-IR processes were densely distributed across the inner plexiform layer (IPL) with a maximum density over lamina 2 of the IPL. Immunoreactive processes also crossed the INL and ramified in the outer plexiform layer where they formed a sparse meshwork. NK1-IR processes were rarely observed in the optic nerve fiber layer. Double-label immunofluorescence studies with different histochemical markers for bipolar cells indicated that NK1 immunoreactivity was not present in bipolar cells. Together, these observations indicate that NK1 immunoreactivity is predominantly expressed by amacrine, displaced amacrine, interplexiform, and some ganglion cells. Double-label immunofluorescence experiments were also performed to characterize NK1-containing amacrine cells. Sixty-one percent of the gamma-aminobutyric acid (GABA)-IR cells, 71% of the large tyrosine hydroxylase (TH)-IR cells, and 100% of the small TH-IR cells contained NK1 immunoreactivity. In addition, most (91%) of the NK1-IR cells had GABA immunoreactivity. In contrast, vasoactive intestinal polypeptide-, TK-, choline acetyltransferase-, and parvalbumin-IR amacrine tells did not express NK1 immunoreactivity. Overall, the present findings suggest that SP acts directly upon several cell populations, including GABA-containing amacrine cells and ganglion cells, to influence visual information processing in the inner retina. PMID- 9414011 TI - A PET study on cortical and subcortical control of pelvic floor musculature in women. AB - The pelvic floor musculature plays an important role in behaviors such as defecation, micturition, mating behavior, and vomiting. A recent positron emission tomography (PET) study revealed that structures belonging to the emotional motor system are involved in the control of the pelvic floor during micturition. However, there also exist brain structures involved in the voluntary motor control of the pelvic floor, and the present PET study was designed to identify these structures. Six adult female volunteers were scanned with the bolus injection of H2(15)O during the following four conditions: (1) rest, (2) repetitive pelvic floor straining, (3) sustained pelvic floor straining, and (4) sustained abdominal straining. The results revealed that the superomedial precentral gyrus, the most medial portion of the motor cortex, is activated during pelvic floor contraction and the superolateral precentral gyrus during contraction of the abdominal musculature. In these conditions, significant activations were also found in the cerebellum, supplementary motor cortex, and thalamus. The right anterior cingulate gyrus was activated during sustained pelvic floor straining. No activations were found in subcortical structures belonging to the emotional motor system. The results are discussed in light of the existing literature on human control of the pelvic floor and micturition. PMID- 9414012 TI - Effect of cocaine self-administration on striatal dopamine D1 receptors in rhesus monkeys. AB - An array of evidence indicates that long-term exposure to cocaine alters several components of the brain dopamine system. Because the release of dopamine in the nucleus accumbens (NAc) has been implicated in mediating the reinforcing effects of cocaine, changes in dopamine function can have profound effects on drug seeking and drug-taking behavior. The present study examined the effects of the chronic self-administration of cocaine on the D1 family of dopamine receptors in the rhesus monkey. The brains of three rhesus monkeys that had intravenously self administered an average of 1.35 mg/kg cocaine per day for 18-22 months were compared to the brains of three cocaine-naive controls. The in vitro quantitative autoradiographic technique was used to quantify binding densities of the D1 ligand [3H]SCH-23390 on cryostat-cut sections of fresh frozen tissue. In animals that self-administered cocaine, the density of D1 binding was significantly lower in the regions of the striatum at the level where the nucleus accumbens is most fully developed. The shell of the NAc showed the largest difference with significantly lower D1 binding also detected in adjacent regions of the caudate nucleus and the putamen. No differences were found in the rostral pole of the NAc or the dorsal striatum at that level. These findings suggest that chronic self administration of cocaine can modulate the density of dopamine D1 receptors in specific portions of the primate striatum. Such changes might underlie some of the behavioral consequences, like drug dependence and craving, of long-term cocaine use. PMID- 9414013 TI - Layer V neurons bear the majority of mRNAs encoding the five distinct dopamine receptor subtypes in the primate prefrontal cortex. AB - In situ hybridization histochemistry was used to determine the laminar distribution of D1, D2, D3, D4, and D5 dopamine receptor mRNAs in the primate prefrontal cortex and to compare striatal and cortical levels of these messages within the same tissue sections. All five subtypes of dopamine receptor mRNA are present in both the monkey striatum and the cerebral cortex but in different proportions within each structure. Thus, levels of D1 and D2 mRNAs are noticeably stronger in the striatum than in the cortex, whereas D4 and D5 expression is clearly higher in the cortex. The D3 transcripts appear nearly equivalent in the striatum and the cortex. A major finding is that, within the prefrontal cortex, mRNAs encoding all dopamine receptor subtypes are expressed most strongly in layer V. This laminar pattern of mRNA distribution does not hold in all cortical regions. The relatively high levels of mRNAs encoding known dopamine receptor subtypes in the primate cerebral cortex, including the D4 receptor, underscore the importance of this structure as a target for therapeutic actions of antipsychotic drugs. Further, their prominence in layer V of the prefrontal cortex, which contains the corticostriatal and corticotectal projection neurons, provides a neural basis for dopaminergic regulation of the descending control systems. PMID- 9414014 TI - Alpha-noradrenergic receptor modulation of the phencyclidine- and delta9 tetrahydrocannabinol-induced increases in dopamine utilization in rat prefrontal cortex. AB - The noncompetitive NMDA receptor antagonist phencyclidine (PCP) and the neuronal cannabinoid receptor agonist delta9-tetrahydrocannabinol (THC) are two agents shown to have psychotomimetic properties in humans. Both drugs increase dopamine release and utilization in the prefrontal cortex, a brain region thought to be dysfunctional in schizophrenia. In the present series of studies, the effects of drugs acting at alpha-noradrenergic receptors on PCP- and THC-induced increases in prefrontal cortical and nucleus accumbens dopamine utilization in the rat were examined. Clonidine, an alpha2 noradrenergic receptor agonist, completely blocked the activation of mesoprefrontal dopamine system by THC or PCP. In addition, the alpha1 noradrenergic receptor antagonist prazosin blocked the PCP-induced increase in prefrontal cortical dopamine utilization. These data may provide new insights concerning pharmacological therapies for acute drug-induced psychoses and behavioral abnormalities in human PCP and THC abusers. PMID- 9414015 TI - Effects of intrapallidal cannabinoids on rotational behavior in rats: interactions with the dopaminergic system. AB - The effect of unilateral intrapallidal cannabinoid receptor stimulation on rotational behavior in rats was explored. The potent cannabinoid agonist CP55,940 (5 microg/0.5 microl) induced ipsilateral turning when microinjected unilaterally into the globus pallidus. The D2 dopamine agonist quinpirole reversed this ipsilateral rotation but failed to affect motor behavior on its own. Finally, the D1 dopamine agonist SKF 82958 inhibited movement when administered into the globus pallidus, and this effect was not additive with CP55,940. PMID- 9414016 TI - Human brain serotonin synthesis capacity measured in vivo with alpha-[C-11]methyl L-tryptophan. AB - Local cerebral serotonin synthesis capacity was measured with alpha-[C-11]methyl L-tryptophan ([C-11]AMT) in normal adult human brain (n = 10; five males, five females; age range, 18-38 years, mean 28.3 years) by using positron emission tomography (PET). [C-11]AMT is an analog of tryptophan, the precursor for serotonin synthesis, and is converted to alpha-[C-11]methyl-serotonin ([C-11]AM 5HT), which is trapped in serotonergic neurons because [C-11]AM-5HT is not degraded by monoamine oxidase. Kinetic analysis of [C-11] activity in brain after injection of [C-11]AMT confirmed the presence of a compartment with unidirectional uptake that represented approximately 40% of the activity in the brain at 50 min after tracer administration. The undirectional rate constant K, which represents the uptake of [C-11]AMT from the plasma to brain tissue followed by the synthesis and physiologic trapping of [C-11]AM-5HT, was calculated using the Patlak graphic approach on a pixel-by-pixel basis, thus creating parametric images. The rank order of K values for different brain regions corresponded well to the regional concentrations of serotonin in human brain (P < .0001). High serotonin synthesis capacity values were measured in putamen, caudate, thalamus, and hippocampus. Among cortical regions, the highest values were measured in the rectal gyrus of the inferior frontal lobe, followed by transverse temporal gyrus; anterior and posterior cingulate gyrus; middle, superior, and inferior temporal gyri; parietal cortex; occipital cortex, in descending order. Values in women were 10-20% higher (P < .05, MANOVA) throughout the brain than those measured in men. Differences in the serotonin synthesis capacity between men and women measured in this study may reflect gender differences of importance to both normal and pathologic behavior. This study demonstrates the suitability of [C 11]AMT as a tracer for PET scanning of serotonin synthesis capacity in human brain and provides normal adult values for future comparison with patient groups. PMID- 9414017 TI - Autoradiographic localization of neurotransmitter binding sites in the hypoglossal and motor trigeminal nuclei of the rat. AB - The hypoglossal and motor trigeminal nuclei contain somatic motoneurons innervating the tongue, jaw, and palate. These two cranial motor nuclei are myotopically organized and contain neurotransmitter binding sites for thyrotropin releasing hormone, substance P, and serotonin. Quantitative autoradiography was used to localize thyrotropin-releasing hormone, substance P, and serotonin-1A and serotonin-1B binding sites in the hypoglossal and motor trigeminal nuclei and to relate the relative distributions of these binding sites to the myotopic organizations of the two nuclei. In the hypoglossal nucleus, high-to-moderate concentrations of all four binding sites were present in the dorsal and ventromedial subnuclei, whereas low concentrations were noted in the ventrolateral subnucleus. In the motor trigeminal nucleus, high concentrations of serotonin-1B, moderate densities of thyrotropin-releasing hormone, and low levels of substance P and serotonin-1A binding sites were present in both the ventromedial and dorsolateral subnuclei. These observations demonstrate that neurotransmitter binding sites in the hypoglossal and motor trigeminal nuclei are heterogeneously localized and that their distributions correspond to the previously described myotopic organizations of each nucleus. PMID- 9414018 TI - Effects of pharmacologic increases in brain GABA levels on cocaine-induced changes in extracellular dopamine. AB - Cocaine-induced increases in extracellular dopamine (DA) concentrations were measured using in vivo microdialysis techniques in the nucleus accumbens (NACC) of freely moving rats. In control animals, cocaine increased extracellular DA concentrations approximately 482% 60 min following administration, returning to baseline values 200 min later. When administered 2 h following an acute dose of gamma-vinyl-GABA (GVG, Vigabatrin), cocaine-induced increases in extracellular DA were reduced to approximately 365% of baseline values. Chronic GVG administration further dose-dependently attenuated the effects of cocaine but did not alter the rate of increase or the rate of return to baseline values. These results indicate that GVG, a drug that increases brain GABA concentrations, is effective in attenuating the effect of cocaine on NACC DA. Taken with our earlier findings, these results support the targeting of brain GABAergic systems as a potentially effective pharmacologic treatment strategy for cocaine addiction. PMID- 9414019 TI - Long-term stability of neurotransmitter activity investigated with 11C-raclopride PET. AB - There is evidence for the shift of regulatory setpoints of functionally linked neurotransmitter systems as a basis of psychiatric disorders. 11C-raclopride PET, which has been shown to be sensitive to changes in endogenous dopamine and has a high short-term test-retest reliability, can be used to investigate different regulatory states of the dopaminergic system with respect to psychiatric diseases and pharmacological influences. Prior to these studies, the reliability of the method over time has to be established. The current study was performed in order to evaluate the long-term stability of the striatal dopaminergic system. Eight normal healthy subjects (mean age: 48.1 years; range: 24-75) were studied twice with 11C-raclopride PET two times under resting conditions with a mean time interval between the scans of 11.3 months (range: 1-19). The ratio of basal ganglia (BG) to cerebellar (CB) distribution volumes (DVs) revealed a mean absolute change of 6.94 (range: 0.0-12.87%) between study A and B. BG DVs mean absolute change was 6.30% (range: 0.55-30.46%), CB DVs mean absolute change was 8.65% (range: 3.51-16.33%). The mean change of the BG/CB ratio was -0.33% (range: 12.87-12.34%). BG DVs mean change was 4.55% (range: 4.2-30.46%), CB DVs mean change was 5.10% (range: -10.71-16.33%). The intraindividual differences between the two scans in our study were not significantly different as compared to the 24 hour interval test-retest data, which have been published earlier (repeated measures ANOVA with df = 11; F = 0.49; P = 0.50) [Volkow et al. (1993) J. Nucl. Med., 34:609-613]. The intraclass correlation of the DV ratio index was r = 0.81. The binding potential in the baseline scans and repeated scans showed a non significant correlation with age (r = -0.58, P = 0.13). Interindividually, the DV ratio index revealed a mean of 3.18 (range = 2.55-3.68, SD = 0.42 in study A and of 3.16 (range 2.37-3.57, SD = 0.41) in study B. The intrasubject stability of the 11C-raclopride binding over a long-term period in normal human subjects suggests the feasibility of study designs investigating the long-term changes of the dopaminergic responsivity after pharmacological challenges. The baseline stability will also serve as a necessary reference for further dose-response studies and investigations of subchronical pharmacological interventions. PMID- 9414020 TI - Interaction between the serotonergic, dopaminergic, and glutamatergic systems in fenfluramine-induced Fos expression in striatal neurons. AB - Fenfluramine (FE) is a halogenated amphetamine derivative used in the treatment of obesity and thought to induce serotonin (5-HT) release from nerve terminals and to reduce re-uptake. However, other pathways may also be involved. In this work, the effects of FE on the major striatal afferent systems, and the possible interactions of these systems in FE-induced striatal expression of Fos, were studied by lesion of the serotonergic and/or dopaminergic system and administration of NMDA glutamate (MK-801) or D1 dopamine (SCH-23390) receptor antagonists. Both the D1 and NMDA receptor antagonists suppressed Fos expression in response to FE almost entirely. FE-induced Fos expression was also dramatically reduced 24 h after 6-hydroxydopamine (6-OHDA) lesion of the dopaminergic system. However, the reduction was not so marked after chronic 6 OHDA lesion, probably due to compensatory changes. Chronic (5,7 dihydroxytryptamine injection, 4 weeks before) or acute (p-chlorophenylalanine injection) lesion of the serotonergic system led to a marked reduction in Fos expression in response to FE (decrease of about 50%). After simultaneous chronic lesion of both serotonergic and dopaminergic systems, a considerable number of Fos-positive nuclei were still observed (decrease of about 70% in the dorsal and dorsomedial regions). The FE-induced expression of Fos was almost totally suppressed (decrease of about 95% in the dorsal and dorsomedial regions) after simultaneous acute lesion. Our results indicate that FE-induced striatal expression of Fos is due in large measure to DA release and dopaminergic stimulation of D1 receptors. However, concurrent stimulation of NMDA glutamate receptors also appears to be essential, and 5-HT release (although not indispensable) doubles striatal Fos expression. PMID- 9414021 TI - High-resolution scatchard analysis shows D1 receptor binding on pyramidal and nonpyramidal neurons. AB - In order to better understand the mechanism of action of atypical antipsychotic drugs (APDs), it is important to clarify how the dopamine system is integrated within local corticolimbic circuits. Toward this end, a high-resolution (HR) Scatchard technique has been used to measure the relative density (Bmax) and affinity (Kd) of D1 receptors on large neurons (> 100 microm2), on small neurons (< 100 microm2), and in neuropil (NPL) of rat medial prefrontal cortex (mPFC) and to determine the laminar distribution of these receptors for each neuronal compartment. Using [3H] SCH23390 as a ligand, all Kd and Bmax values were found to be similar indicating that D1 receptor activity is not preferentially localized to either large or small neuronal subtypes in mPFC. The density of D1 receptor binding in all three compartments was found to be almost twice as great in layers V and VI, as compared to superficial layers II and III. These results suggest that the blockade of D1 receptors associated with some atypical APDs may involve both pyramidal and nonpyramidal neurons in the PFC. PMID- 9414022 TI - Synchronous firing of inhibitory interneurons results in saturation of fast GABA(A) IPSC magnitude but not saturation of fast inhibitory efficacy in rat neocortical pyramidal cells. AB - The kinetic properties of evoked fast inhibitory postsynaptic currents were examined to elucidate factors underlying the limit on the magnitude of fast inhibition in neocortex. Using whole-cell voltage-clamp recordings from layer V pyramidal neurons in slices of rat somatosensory cortex, fast gamma-aminobutyric acid-A (GABA[A])ergic inhibitory postsynaptic currents were selectively recorded by holding cells at potentials equal to excitatory postsynaptic current reversal (approximately 0 mV). As stimulus intensity was increased, the magnitude and duration of the fast inhibitory postsynaptic current increased. Over the range of stimuli applied (2-10 V), fast GABA(A)-mediated inhibitory postsynaptic currents reached a maximum peak conductance of 25.9 +/- 4.2 nS (range 10.5-41.2 nS) at intensities approximately 2-times threshold. As stimulus intensities were increased beyond this point of maximal conductance, the time constant of current decay increased as function of stimulus strength, while rise time remained unaffected. Exposure to nominally magnesium-free solutions did not affect maximal peak conductances of fast inhibitory postsynaptic currents, but did cause an increase in the time constants of current decay by 66.3 +/- 23.6%, resulting in an 85.6 +/- 24.6% increase in the total charge flux carried by single inhibitory postsynaptic currents. This effect may be due to prolonged activation of postsynaptic GABA(A) receptors by excess GABA released in response to increased excitation. Exposure to the GABA uptake blocker, nipecotic acid, similarly prolonged current decay without affecting the maximal peak conductance. Our findings suggest that the limit on recruitment of evoked fast inhibition in neocortical layer V pyramidal cells arises from the saturation of postsynaptic GABA(A) receptors. However, while there is a limit to the peak fast inhibitory postsynaptic conductance which can be recruited with increasing excitation, inhibitory strength may still be modulated by increasing charge flux through the prolongation of fast inhibition. PMID- 9414024 TI - Significant roles of inducible cyclooxygenase (COX)-2 in angiogenesis in rat sponge implants. AB - Angiogenesis in rat sponge implants, as determined from the concentration of hemoglobin in the sponge granuloma tissues, was gradually increased over a 14-day experimental period. The inducible cyclooxygenase COX-2 was detected in the sponge granuloma tissues at day 4 by Western blot analysis using specific mouse COX-2 antibody. Angiogenesis in the sponge implants was enhanced by daily topical injections of human recombinant basic fibroblast growth factor (bFGF) or human recombinant epidermal growth factor (EGF) (100 or 1000 ng/sponge/day) for 4 days. These treatments clearly enhanced the expression of COX-2 in the sponge granuloma tissues. In immunohistochemical studies, COX-2-positive staining was mainly observed in the endothelial cells of the neovasculature and in the fibroblasts of the granuloma capsule. Administration of the selective COX-2 inhibitor NS-398 (p.o., 3 mg/kg, 3 times a day) for 14 days significantly inhibited the angiogenesis. The angiogenesis enhanced with bFGF or EGF (day 4) was inhibited by administration of indomethacin or NS-398, both in the above regimen, and fell to the level obtained without growth factor treatment. These results suggest that COX-2 induced in the sponge granuloma tissues may participate in neovascularization through prostaglandin formation. PMID- 9414023 TI - Acute effects of various GABA receptor agonists and glutamate antagonists on focal hippocampal seizures in freely moving rats elicited by low-frequency stimulation. AB - In this study, we examined the acute anticonvulsant spectrum of (1) dizocilpine (0.03-3 mg/kg), CGS 19755 (1-10 mg/kg), and 7-chlorokynurenic acid (1-100 nmol) (NMDA receptor/ionophore complex antagonists); (2) muscimol (0.1-10 nmol; direct GABA(A) agonist); (3) YM90K (3-10 mg/kg; AMPA receptor antagonist); and (4) diazepam (2 and 5 mg/kg) and carbamazepine (5 and 20 mg/kg), two standard anticonvulsants, using the partially-kindled hippocampal model for epileptic seizures in freely moving rats. The anticonvulsant effect of these compounds were assessed by determining (1) the afterdischarge (AD), which is indicative of the severity of the seizure and related to seizure maintenance, and (2) the pulse number threshold (PNT), which is indicative of the seizure threshold or initiation. In addition, ataxia, a measure of CNS dysfunction, was assessed for each compound. Overall, our results indicated that the anticonvulsant compounds examined could be classified into three categories based on effects on the AD and PNT: (1) elevation of PNT (carbamazepine, dizocilpine, CGS 19755 and 7 chlorokynurenic acid); (2) reduction of AD (diazepam and muscimol); and (3) mixed action, i.e., increased PNT and decreased AD (YM90K). Behavioral data indicated that all compounds, except carbamazepine, produced a dose- or concentration dependent ataxia. Overall, our results suggest that NMDA receptors play a role in seizure initiation, whereas the GABA(A) receptors appear to be involved in seizure maintenance and AMPA receptors may be involved in both phenomena. PMID- 9414025 TI - Histamine-induced cortisol secretion from bovine adrenocortical cells: co incubated with bovine adrenal medullary cells. AB - Histamine at concentrations higher than 10(-9) M significantly elicited cortisol secretion from bovine adrenocortical (BAC) cells co-incubated with bovine adrenal medullary (BAM) cells, suggesting that BAM cells are responsible for histamine induced cortisol secretion. Cortisol secretion from BAC cells co-incubated with BAM cells was also elicited by both an H1 agonist, 2-methylhistamine, and an H2 agonist, 4-methylhistamine. However, 4-methylhistamine was much less effective than 2-methylhistamine. Histamine-induced cortisol secretion was inhibited not only by H1 antagonists (pyrilamine and diphenhydramine) but also by H2 antagonists (cimetidine and ranitidine). Histamine effectively increased 45Ca uptake and IP3 production in BAM cells. These responses were antagonized by the H1 antagonist but not by the H2 antagonist. Histamine-induced cortisol secretion from BAC cells co-incubated with BAM cells was inhibited by beta-adrenoceptor antagonists, propranolol and timolol, as well as an NK1-receptor antagonist, D Arg1-D-Trp7,9-Leu11-substance P. These results indicate that histamine can induce cortisol secretion from BAC cells at physiological concentrations through H1 receptors on BAM cells, and catecholamine and substance P may participate in histamine-induced cortisol secretion. PMID- 9414026 TI - Inhibition by nitric oxide of the uptake of [3H]serotonin into rat brain synaptosomes. AB - [3H]Serotonin (5-HT) uptake by synaptosomes of rat brain was dose-dependently inhibited by nitric oxide (NO) donors such as sodium nitroprusside (SNP), 3-(2 hydroxy-1-methyl-2-nitrosohydrazino)-N-methyl-1-propanamin e, 3 morpholinosydnonimine and S-nitroso-L-cysteine (NO-CYS). The inhibitory effect was blocked by reduced hemoglobin. The effect was not mimicked by ferrocyanide and ferricyanide. 8-Bromoguanosine 3',5'-cyclic monophosphate (8-bromo cGMP) did not affect [3H]5-HT uptake into rat cortical synaptosomes. The reduced activity of [3H]5-HT uptake into the cortical synaptosomes pretreated with NO-CYS was partially reversed by washing the preparation after the treatment. Kinetic analysis showed that NO-CYS (100 microM) decreased the Vmax value without any change in the Km value. NO-CYS did not affect the specific binding of [3H]paroxetine, a ligand that binds to the 5-HT transporter, in membranes. NO-CYS and SNP, like iodoacetic acid and sodium cyanide, decreased the ATP content in cortical synaptosomes, but the effect on ATP content was not related to that on [3H]5-HT uptake. These findings suggest that NO inhibits reversibly [3H]5-HT uptake into rat brain synaptosomes without affecting the recognition site of the 5-HT transporter in a cGMP-independent manner, and the observed effect is not due to its metabolic effect. PMID- 9414027 TI - Suppressive effects of Y-24180, a long-acting antagonist for platelet-activating factor, on allergic pulmonary eosinophilia in mice. AB - We studied the effects of (+/-)-4-(2-chlorophenyl)-2-[2-(4-isobutylphenyl)ethyl] 6,9-dimethy l-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepine (Y-24180), a long-acting antagonist for platelet-activating factor (PAF), on antigen-induced eosinophil infiltration and interleukin-5 (IL-5) release in the bronchoalveolar lavage fluid (BALF) of mice. Mice actively sensitized with ovalbumin (OA) were challenged by injecting intratracheally OA 3 times every fourth day. Both the number of eosinophils and level of IL-5 were significantly increased in the BALF 24 hr after the last OA challenge. Either Y-24180 or prednisolone was orally administered once a day for 10 days beginning one day before the first OA challenge. WEB2086, another PAF antagonist, was orally administered once or twice a day for 10 days. Y-24180 (0.3 - 3 mg/kg) suppressed the eosinophil infiltration in a dose-dependent manner and suppressed the IL-5 release at the highest dosage. Prednisolone (10 mg/kg) significantly suppressed both the eosinophil infiltration and IL-5 release. In contrast, WEB2086 affected neither the eosinophil infiltration nor IL-5 release when administered once a day (10 - 100 mg/kg/day). This drug never affected the IL-5 release but significantly suppressed eosinophil infiltration even when administered twice a day (30 - 200 mg/kg/day). These results indicate that the suppressive effect of Y-24180 on allergic pulmonary eosinophilia is due to not only to its long-lasting PAF-antagonism but also due to its suppressive effect on IL-5 release. PMID- 9414028 TI - Effects of vitamin K2 (menatetrenone) on atherosclerosis and blood coagulation in hypercholesterolemic rabbits. AB - Gamma-Carboxyglutamic acid (Gla)-containing protein, synthesized in the presence of vitamin K, has been found in atherogenic plaques, but the pharmacological effect of vitamin K on atherosclerosis is unclear. We examined whether vitamin K2 (menatetrenone) could affect the progression of both atherosclerosis and hypercoagulability in hypercholesterolemic rabbits. Vitamin K2 in daily doses of 1, 10 and 100 mg/kg was given with a 0.5% cholesterol diet for 10 weeks to 8 rabbits each. The plasma levels of total-cholesterol in the vitamin K2-treated groups were clearly lower than that of the hypercholesterolemic control group. The excessive dose of vitamin K2, even at the high dose of 100 mg/kg/day for 10 weeks, did not accelerate the progression of atherosclerosis and did not promote the coagulative tendency in the rabbits. In contrast, the vitamin K2 treatment (1 to 10 mg/kg/day) suppressed the progression of atherosclerotic plaques, intima thickening and pulmonary atherosclerosis, the increase of ester-cholesterol deposition in the aorta, and both the elevation in plasma factor X level and increase in Hepaplastin test value in the rabbits. These results indicate that the pharmacological dose of vitamin K2 prevents both the progression of atherosclerosis and the coagulative tendency by reducing the total-cholesterol, lipid peroxidation and factor X activity in plasma, and the ester-cholesterol deposition in the aorta in hypercholesterolemic rabbits. PMID- 9414029 TI - Low sensitivity of adrenal chromaffin cells to acetylcholine, neostigmine and oxotremorine in 21-day-old rats. AB - In the previous study, we have demonstrated that nicotine hardly induces catecholamine release from adrenal medulla of 21-day-old rats. The present study examined the responsiveness of the adrenal chromaffin cells to acetylcholine in vitro and neostigmine and oxotremorine in vivo in 21-day-old and 8-week-old rats. As assessed by electron microscopy, the number of the chromaffin granules was markedly decreased and the content of adrenaline in adrenals was diminished significantly by oxotremorine treatment in 8-week-old rats, whereas these changes did not occur in 21-day-old rats. Morphological changes of the adrenal chromaffin cells, with respect to exocytosis, were not observed in neostigmine-treated 21 day-old rats and acetylcholine-treated adrenal slices prepared from 21-day-old rats. Catecholamine release was hardly evoked by acetylcholine in these slices as judged by measuring the catecholamine content in the medium. These results indicate that the sensitivity of the chromaffin cells to these secretagogues in 21-day-old rats is very low when compared to that in young adult rats. PMID- 9414030 TI - Rolipram, a phosphodiesterase-4-selective inhibitor, promotes the survival of cultured rat dopaminergic neurons. AB - We evaluated the effects of rolipram, a selective inhibitor of phosphodiesterase (PDE) 4, on the survival of dopaminergic neurons in 13-day culture. Rolipram did not affect the survival of dopaminergic neurons in the absence of forskolin, but significantly enhanced the survival of dopaminergic neurons in the presence of 10(-5) M forskolin in a concentration-dependent manner (10(-8) - 10(-5) M). Rolipram also enhanced the neurotrophic effect of forskolin on total neurons including dopaminergic and non-dopaminergic neurons at a high concentration (10( 5) M), but did not affect the survival of cells containing glutamate or gamma aminobutylic acid. A non-selective PDE inhibitor, 1-isobutyl-3-methylxanthine, caused a marked increase of dopaminergic neurons, whereas selective inhibitors of PDE2 and PDE3 showed far weaker effects. A PDE1 inhibitor, on the other hand, caused non-specific cell death in the presence or absence of forskolin. These findings suggest that rolipram has a potential to enhance the survival of dopaminergic neurons selectively by way of PDE4 inhibition. PMID- 9414031 TI - A modified coaxial compound micropipette for extracellular iontophoresis and intracellular recording: fabrication, performance and theory. AB - Investigation of the identity and modes of action of neurotransmitters in the mammalian central nervous system can be facilitated by simultaneous intracellular recording of membrane potential and extracellular iontophoresis of agonists and antagonists. We describe here techniques for conveniently constructing a compound microelectrode, originally described by Sonnhof (Pflugers Arch 341, 351-358, 1973), suitable for such studies. The Sonnhof electrode consists of two components, a centraxial micropipette for recording membrane potential surrounded by a cylindrical array of 6 pipettes for iontophoresis. The cylindrical array tapers coaxially and terminates in 6 contiguous, crescent-shaped orifices surrounding the terminal portion of the central pipette, 25 - 50 microm from the tip. Pipettes were constructed from borosilicate glass tubing of 1-mm wall thickness having a 10-mm or 16-mm outer diameter. The resistances, flux and transport numbers for iontophoresis of glycine were measured for pipettes constructed from both sizes of glass. Flux increased with increasing levels of current, and transport number decreased with increasing micropipette resistance. A spherical diffusion model points out the steep dependence of steady state concentration on diffusional distance, stressing the importance of diminishing the distance between the iontophoresis source and the recording site. This is particularly true when brief pulses of current are used. PMID- 9414032 TI - Receptor subtypes involved in the alpha1-adrenoceptor mediated increase in 86Rb+ efflux from the rat heart. AB - The aim of this study was to determine the involvement of the different alpha1 adrenoceptor subtypes in the alpha1-adrenoceptor mediated increase in 86Rb+ efflux from rat hearts. Isolated hearts were perfused in the presence of a beta adrenoceptor antagonist (1 microM timolol). After loading with 86Rb+, the efflux was measured during alpha1-adrenoceptor stimulation by phenylephrine (30 microM). Phenylephrine increased the 86Rb+ efflux by about 30%. Pretreatment with the preferentially alpha1B-adrenoceptor inhibitor chloroethylclonidine (CEC), reduced the response to phenylephrine by about 50%. The preferential alpha1D-adrenoceptor inhibitor BMY 7378 inhibited the response to phenylephrine by 35%, with a pKI=8.4 (95% C.I. 8.2-8.6). The response was sensitive to the preferential alpha1A adrenoceptor inhibitors (+)niguldipine, 5-methylurapidil (5-MU) and WB-4101 at relatively high concentrations, and 5-MU inhibited the response with a pKI=7.7 (95% C.I. 7.2-8.0) in CEC pretreated hearts. In conclusion, the phenylephrine stimulated increase in 86Rb+ efflux in the rat heart is not specifically linked to only one of the alpha1-adrenoceptor subtypes, but involves the alpha1B- and the alpha1D-adrenoceptor subtypes, and probably the alpha1A-adrenoceptor subtype as well. PMID- 9414033 TI - Effects of perospirone, a novel antipsychotic agent, on the dopaminergic neurons in the rat ventral tegmental area. AB - An electrophysiological study was performed to investigate the effects of cis-N [4-[4-(1,2-benz-isozole-3-yl)-1-piperazinyl]butyl]cyclohexan e-1,2-dicarboximide hydrochloride (perospirone), a novel antipsychotic agent with high affinities for D2/5-HT2-receptors, on the dopaminergic (DA) neurons in the ventral tegmental area (VTA) using chloral hydrate-anesthetized rats. DA neurons and non-DA neurons in VTA were identified according to the configurations of their action potentials and firing rates. Spontaneous firing of DA neurons was dose-dependently decreased by i.v. injection of methamphetamine (MAP). Most non-DA neurons were unaffected by MAP up to 2 mg/kg, but the firing was increased with MAP in 2 of 7 neurons. Perospirone injected intravenously reversed the MAP-induced decrease in spontaneous firing of DA neurons in a dose-dependent manner. In addition, i.v. injection of perospirone also inhibited the MAP-induced increase in firing of the 2 non-DA neurons. Similarly, inhibition of spontaneous firing in DA neurons by microiontophoretically applied DA was antagonized during iontophoretic application of perospirone. However, the firing of non-DA neurons, which were insensitive to DA, was not affected by iontophoretically applied perospirone. Since the DA neurons are inhibited by DA via D2-receptors, these findings suggest that perospirone acts on the D2-receptors to antagonize the dopaminergic inhibition of DA neurons in VTA. PMID- 9414034 TI - Inhibition by aminosalicylates of lipid peroxidation in large intestinal mucosa after mesenteric ischemia/reperfusion in the rat. AB - To clarify the mode of action of aminosalicylates, which are generally used as therapeutic agents for ulcerative colitis, we investigated the effect of some of the aminosalicylates on lipid peroxidation in the large intestinal mucosa after mesenteric ischemia/reperfusion in the rat. Lipid peroxidation was assessed by measuring the level of thiobarbituric-acid-reactive substances. It was found that aminosalicylates dose-dependently inhibited the elevation of the level of thiobarbituric-acid-reactive substances in the large intestinal mucosa after ischemia/reperfusion. This effect may partly contribute to the therapeutic actions of aminosalicylates in ulcerative colitis. PMID- 9414035 TI - Comparative study of the effects of indomethacin and NS-398, a selective cyclooxygenase 2 inhibitor, on duodenal bicarbonate secretion induced by luminal acidification in rats. AB - To clarify the mechanisms of duodenal ulcerogenic activity of non-steroidal anti inflammatory drugs (NSAIDs), the effects of indomethacin (IND) on acid-stimulated duodenal bicarbonate secretion and histamine-induced duodenal ulcerogenic responses were studied in comparison with NS-398, a selective cyclooxygenase (COX)-2 inhibitor, in rats. IND (1 and 5 mg/kg, s.c.) significantly decreased duodenal bicarbonate secretion and potentiated duodenal lesion in a dose dependent manner. On the other hand, NS-398 had no effect on these parameters. These findings suggest that duodenal ulcerogenicity of IND in the presence of histamine is mainly due to the inhibitory action on acid-stimulated bicarbonate secretion mediated by COX-1, but not by COX-2. PMID- 9414036 TI - Possible involvement of rapamycin-sensitive pathway in Bcl-2 expression in human neuroblastoma SH-SY5Y cells. AB - In human neuroblastoma SH-SY5Y cells, treatment with immunosuppressants such as FK506, cyclosporin A or rapamycin for 4 days induced the enhancement of the 27 kDa Bcl-2alpha protein level. Among immunosuppressants, rapamycin has most potency. Treatment with herbimycin A or wortmannin also enhanced Bcl-2 expression, but the BB type of platelet-derived growth factor decreased the level. These results suggest that Bcl-2 expression is probably regulated by the cascade of tyrosine kinase, phosphatidylinositol 3-kinase and rapamycin-sensitive p70 S6-kinase in human neuroblastoma SH-SY5Y cells. PMID- 9414037 TI - Effects of concanavalin A on cytokine mRNA expression in mouse liver. AB - The effects of concanavalin A (Con A) on liver cytokine gene expression was studied in mice. The CD4 mRNA expression in normal liver suggests the presence of CD4+ T cells. The administration of Con A induced interleukin (IL)-1beta, IL-2 and IL-2 receptor mRNAs, which implies lymphocyte activation in the liver. Interferon-gamma and tumor necrosis factor-alpha mRNA expressions were increased gradually. The present results showed that Con A induced liver cytokine genes. This cytokine gene induction might have been the result of lymphocyte activation in the liver. PMID- 9414038 TI - Pituitary adenylate cyclase-activating polypeptide (PACAP) receptor mRNA in the rat adrenal gland: localization by in situ hybridization and identification of splice variants. AB - The distribution of mRNA for pituitary adenylate cyclase-activating polypeptide receptor (PACAP-R) was examined by in situ hybridization in rat adrenal gland. In the adrenal medulla, PACAP-R mRNA was expressed in almost all chromaffin cells without any significant expression in the cortical region. Using the reverse transcription-polymerase chain reaction, we analyzed the mRNA expression of PACAP R splice variant forms in the adrenal gland. The predominant forms observed were the variant having a 28-amino acid insert in the third intracellular loop (termed PACAP-R-hop1). As PACAP is localized in the noradrenaline secreting cells of the adrenal chromaffin cells, and stimulates catecholamine release, the present results suggest that PACAP may serve as a paracrine or autocrine regulatory factor for the chromaffin cells through PACAP-R. PMID- 9414040 TI - Treatment and prognosis of gastric lymphoma. AB - To assess the developments in the prognosis and treatment of patients with primary gastric non-Hodgkin's lymphoma we reviewed 178 papers in English, German, and French on the subject that were listed on MEDLINE between January 1974 and April 1995. We analysed the results of 3157 patients, and during that period the overall survival increased from 37% to 87%. Overall survival by Ann Arbor stage was 57%. 998/1296 (77%) for stage IE, 231/330 (70%) for stage II1E, 107/290 (37%) for stage II2E, 53/172 (31%) for stage IIIE, and 122/451 (27%) for stage IV. Over half the publications recommended resection alone. Only 12 (15%) thought that radiotherapy or chemotherapy, alone or in combination, was adequate. The results of all treatments were similar in 1296 patients with stage IE disease. For stage IIE-IVE disease, however, 66 (82%) of authors suggested a treatment protocol that included resection, and of these 39 (49%) recommended a combination of resection, local radiotherapy, and systemic chemotherapy. The number of patients reported was too small for us to be able to give precise recommendations for treatment of gastric non-Hodgkin's lymphoma, and we have been able to give only an evaluation of current treatments. PMID- 9414039 TI - Inhibitory effect of lomerizine, a diphenylpiperazine Ca2+-channel blocker, on Ba2+ current through voltage-gated Ca2+ channels in PC12 cells. AB - We investigated the effect of lomerizine, an anti-migraine drug, on the Ba2+ current through voltage-gated Ca2+ channels in rat pheochromocytoma (PC12) cells using a whole-cell voltage-clamp technique. Lomerizine inhibited the Ba2+ current with an IC50 value of 1.9 microM. Lomerizine and nicardipine were >4 times more potent than flunarizine, diltiazem, verapamil and dimetotiazine. The time course of inactivation induced by lomerizine was similar to that induced by nicardipine and flunarizine. These data indicate that lomerizine may inhibit the Ca2+ channel in a similar manner to nicardipine and flunarizine, and its potency is almost equal to that of nicardipine. PMID- 9414041 TI - Selective avoidance of postoperative locoregional radiotherapy in breast cancer seems to be justified. AB - OBJECTIVE: To try and reduce the amount of routine postoperative radiotherapy that we prescribed without causing an unacceptable rise in locoregional recurrences. DESIGN: Retrospective study. SETTING: Teaching hospital, The Netherlands. SUBJECTS: 836 women who were treated for breast cancer between January 1980 and December 1989. INTERVENTIONS: These 836 had been treated by modified radical mastectomy (n = 534), excision of the tumour and axillary dissection (n = 279), lumpectomy (n = 15), or total mastectomy (n = 8). In December 1984 we stopped giving routine postoperative irradiation to women with T1 or T2 tumours unless there was any doubt about the operative specimen. MAIN OUTCOME MEASURES: The rate of locoregional recurrence 1985-9 compared with that from 1980-December 1984. RESULTS: Only 1 patient of 836 had a clinically detectable recurrence in the internal mammary chain. There were only 2 recurrences in the 235 axillas that had not been irradiated. CONCLUSION: By a process of careful selection of patients for locoregional irradiation, the number of fields of irradiation given to patients with breast cancer can be reduced by up to 80% without causing a rise in the rate of locoregional recurrences. PMID- 9414042 TI - Hernia surgery in a defined population: a prospective three year audit. AB - OBJECTIVE: To establish a register of inguinal hernia surgery that allows audit and analyses of data from several centres. DESIGN: Prospective recording of data on a common protocol. SETTING: Eight Swedish hospitals. SUBJECTS: All groin hernia operations done for patients over 15 years old from January 1992 to December 1994. MAIN OUTCOME MEASURES: Methods of repair, postoperative complications including mortality, day surgery rate, and reoperations for recurrence. RESULTS: During the three years studied 4879 hernia operations were undertaken in 4474 patients. Postoperative mortality within 30 days of operation for emergency and elective hernia repairs was 3.5% and 0.07%, respectively. Of all herniorrhaphies 798 (16%) were done for recurrences, 142 of these after operations between 1992 and 1994. At 24 months 4% of all operations had been redone because of recurrences with highly significant variations among hospitals ranging from 1.5% to 6.7%. Postoperative complications within 30 days after operation, direct hernia, recurrent hernia, and the use of absorbable sutures were associated with an increased risk of reoperation. CONCLUSIONS: A quality register recorded voluntarily can identify significant interhospital differences in outcome as well as variables associated with an increased risk of reoperation, thereby raising quality awareness and facilitating the process of improvement. PMID- 9414043 TI - Diagnostic decision support in suspected acute appendicitis: validation of a simplified scoring system. AB - OBJECTIVE: To validate a simplified scoring system as an aid to the diagnosis of acute appendicitis. DESIGN: Open prospective study. SETTING: County district hospital, and university hospital, Sweden. SUBJECTS: 1167 Patients with suspected appendicitis. MAIN OUTCOME MEASURES: Correlation between scoring system and final diagnosis. RESULTS: A total of 475 patients were operated on and 392 (82.5%) of these had histologically verified appendicitis. The negative laparotomy rate was 17.5% (11.2% for men and 25.4% for women). The sensitivity of the scoring system for appendicitis at the main cut-off point (score -2 or more) was 0.73 and the specificity was 0.87. At the cut-off level (score - 17 or less) for predicting non-specific abdominal pain (NSAP) the proportion of correctly classified patients was 0.72 and the proportion of false negatives (patients with appendicitis classified as NSAP) was 0.14. Analysis of the area under the receiver operating characteristic (ROC) curve showed that the scoring system performed slightly worse in the university hospital (area 0.83) than in the district hospital where it was originally developed (area 0.89). CONCLUSION: The scoring system was a valid instrument for discriminating between acute appendicitis and NSAP in the two centres studied. Use of the scoring system in daily clinical work was associated with a reduced rate of negative laparotomies. PMID- 9414044 TI - Y-V anoplasty combined with internal sphincterotomy for stenosis of the anal canal. AB - OBJECTIVE: To review the results of Y-V anoplasty combined with internal sphincterotomy in the treatment of anal stenosis. DESIGN: Retrospective study. SETTING: University hospital, Finland. SUBJECTS: 10 patients with stenosis of the anal canal. MAIN OUTCOME MEASURES: Improvement of symptoms and function. RESULTS: Nine patients improved postoperatively. Six of the patients had good results, three had fair results and one had a poor result. There were no postoperative complications. CONCLUSION: Y-V anoplasty combined with internal sphincterotomy is a safe and simple procedure that gives good results. We recommend its use to treat stenosis of the anal canal. PMID- 9414045 TI - An experimental porcine model of partial ischaemia of the distal colon. AB - OBJECTIVE: Ischaemia of the colon is a major challenge in aortoiliac surgery. The aim was to establish an animal model of partial distal colonic ischaemia to study interventional strategies. DESIGN: Randomised experiment. SETTING: University Hospital. Department of Experimental Research. MATERIAL: 19 pigs. INTERVENTIONS: 11 Pigs were subjected to ischaemia consisting of total occlusion of the inferior mesenteric artery and partial occlusion of the superior mesenteric artery. Eight animals were sham controls. Dextran was given. MAIN OUTCOME MEASURES: Haemodynamic measurements, intramucosal pH-measurements (pHi) and histological grading. RESULTS: Both ischaemic animals and controls remained haemodynamically stable. It was possible to maintain stable ischaemia in the distal colon in the pHi-range of 6.9-7.1. There was histological mucosal damage of the distal colon in ischaemic animals but not in controls. CONCLUSIONS: The model could be of value when studying interventional strategies to reduce or reverse ischaemia. PMID- 9414046 TI - Impairment of bone and mineral metabolism after pancreatic transplantation: observations in syngeneic rats. AB - OBJECTIVE: To find out whether transplantation of the duodenum and pancreas results in osteopathy in immunotolerant recipient rats. DESIGN: Prospective study. SETTING: University hospital, Germany. ANIMALS: 30 male Lewis rats weighing about 315 g. INTERVENTIONS: Laparotomy alone (n = 10, control), or transplantation by one of three techniques: ectopic extraportal (EE, n = 5), ectopic intraportal (EI, n = 7), or orthotopic intraportal (OI, n = 8). MAIN OUTCOME MEASURES: Body weight; changes in mineral and bone metabolism, bone density and ash, and the breaking force of bone. RESULTS: Rats were followed for 133 plus or minus 3 days. Compared with the control group the rats that had undergone transplantation had delayed gain in body weight and the final weight was reduced. Urinary glucose and basal serum glucose and insulin concentrations were unaffected, but that of pancreatic polypeptide was low. In the transplanted groups intestinal absorption of calcium and phosphorus and serum concentrations of calcium and parathyroid hormone were within reference ranges. Serum magnesium (EI and OI) and phosphorus (EI) concentrations, and the urinary calcium (FE) and phosphorus (EI) concentrations, and the urinary hydroxyproline:serum osteocalcin ratio (FE, and EI) were significantly higher than in controls (p < 0.01). CONCLUSIONS: Transplantation failed to disturb glucometabolic functions and intestinal mineral absorption but was associated with an osteopathy which showed itself in high bone resorption and reduced bone phosphorus and magnesium concentrations. The osteopathy developed irrespective of the technique of reconstruction of the digestive tract and so may be caused by division of intestinal nerves or their incomplete regeneration. PMID- 9414047 TI - Interstitial laser thermotherapy of adenocarcinoma transplanted into rat liver. AB - OBJECTIVE: To examine the effect of different temperatures and exposure times in interstitial laser thermotherapy. DESIGN: Controlled laboratory study. SETTING: University hospital, Sweden. MATERIAL: 48 male Wistar FU rats with dimethylhydrazine-induced adenocarcinoma transplanted into the liver. INTERVENTION: Treatment was given with an Nd:YAG laser and a feedback system for temperature regulation. Light was delivered into the centre of the tumour and the feedback thermistor probe was placed 3 mm from the tumour margin. Rats were treated at steady-state temperatures at the feedback thermistor of 43, 46, or 50 degrees C for 30 minutes, and at a steady-state temperature of 46 degrees C at the feedback thermistor also for 10 and 20 minutes. MAIN OUTCOME MEASUREMENT: Tumour control as assessed 6 days after treatment using light microscopical examination including immunohistochemical determination of bromodeoxyuridine (BrdU) incorporation into DNA as a measure of cell viability. RESULTS: Complete tumour necrosis was achieved in all rats treated for 30 minutes, in 6/8 rats treated for 10 minutes and in 6/8 rats treated for 20 minutes at 46 degrees C. During steady-state thermotherapy, temperatures at the tumour margin were about 11 degrees higher than at the feedback thermistor (range 54-61 degrees C). The surrounding liver tissue also became necrotic so that the total necrosis volume exceeded the pretreatment tumour volume. CONCLUSION: Interstitial laser thermotherapy at temperatures ranging from 54-61 degrees C at the tumour margin ensures total necrosis of a transplanted rat liver carcinoma provided that treatment is given for 30 minutes. PMID- 9414048 TI - Carcinoma of the breast presenting as a thyroid mass. PMID- 9414049 TI - Strangulated internal supravesical hernia: a diagnostic problem. PMID- 9414050 TI - The history of pain project. PMID- 9414051 TI - Drug interactions in human neuropathic pain pharmacotherapy. AB - Current practice predicates the use of multiple drug combinations in the treatment of neuropathic pain. These combinations may be required because of multiple pain symptoms directly arising from neuropathic pathology, other symptoms attributable to the chronicity and severity of the patient's pain or conditions unrelated to their pain. A fear exists that combination drug use or the addition of a new drug to a therapeutic regimen may lead to increased drug toxicity or decreased efficacy. Many of the drug interactions of significance to neuropathic pain physicians involve the cytochromes P450 2D6 and 3A3/4 isoenzymes. Drug interactions should be more predictable based on the knowledge of which compounds induce, inhibit or are metabolized by specific cytochrome P450 enzymes. Mechanisms of induction or inhibition of biotransformation via the P450 hepatic enzyme system are discussed and various inducers, inhibitors and substrates relating to neuropathic pain pharmacotherapy are listed. PMID- 9414052 TI - Resolution of psychological distress of whiplash patients following treatment by radiofrequency neurotomy: a randomised, double-blind, placebo-controlled trial. AB - It is well recognised that patients with chronic pain, in particular, chronic whiplash-associated neck pain, exhibit psychological distress. However, debate continues as to whether the psychological distress precedes and causes the chronic pain or, conversely, the psychological distress is a consequence of chronic pain. Using cervical zygapophysial joint pain as a model for chronic pain, the effect of a definitive neurosurgical treatment on the associated psychological distress was studied. Seventeen patients with a single painful cervical zygapophysial joint participated in a randomised, double-blind, placebo controlled trial of percutaneous radiofrequency neurotomy. Their pain and psychological status were evaluated pre-operatively and 3 months post-operatively by medical interview and examination, a visual analogue pain scale, the McGill Pain Questionnaire, and the SCL-90-R psychological questionnaire. All patients who obtained complete pain relief exhibited resolution of their pre-operative psychological distress. In contrast, all but one of the patients whose pain remained unrelieved continued to suffer psychological distress. Because psychological distress resolved following a neurosurgical treatment which completely relieved pain, without psychological co-therapy, it is concluded that the psychological distress exhibited by these patients was a consequence of the chronic somatic pain. PMID- 9414053 TI - Stimulation of the greater occipital nerve increases metabolic activity in the trigeminal nucleus caudalis and cervical dorsal horn of the cat. AB - Patients with primary headache syndromes often describe a distribution of pain that involves both frontal and occipital parts of the head. Such a distribution of pain does not respect the cutaneous sensory innervation of the head which would divide it into anterior (trigeminally innervated) and posterior (spinal nerve root innervated) regions. Studies of pain-producing intracranial structures, such as the superior sagittal sinus, have demonstrated that second order neurons as caudal as C2 are activated after either electrical or mechanical stimulation. For this study cats were anaesthetised with halothane (during surgery) and alpha-chloralose (60 mg/kg, i.p., then 20 mg/kg intravenous maintenance), paralysed (gallamine 6 mg/kg) and ventilated. The greater occipital nerve was isolated bilaterally and stimulated unilaterally using hook electrodes with stimuli of 100 V at 0.3 Hz. Metabolic activity in the caudal brain stem and upper cervical cord was measured using 2-deoxyglucose autoradiography and quantitative densitometry. Stimulation of the greater occipital nerve increased metabolic activity by 220% ipsilateral to stimulation and by a lesser amount contralaterally. Increases in metabolic activity were seen in the dorsal horn at the level of C1 and C2 as might be predicted from the cervical origin of the nerve. Neuronal activation appeared contiguous with the trigeminal nucleus caudalis and was in the same distribution as is seen when trigeminally-innervated structures are stimulated. These data suggest that the well recognised clinical phenomenon of pain at the front and back of the head and in the upper neck are likely to be a consequence of overlap of processing of nociceptive information at the level of the second order neurons. PMID- 9414054 TI - Non-specific musculoskeletal pain in preadolescents. Prevalence and 1-year persistence. AB - A 1-year follow-up study of 1756 third- and fifth-grade schoolchildren was conducted with a structured pain questionnaire to assess the prevalence and persistence of self-reported musculoskeletal pain symptoms and disability caused by pain. At follow-up, 1626 (92.7%) children participated in the study. Pain at least once a week persisted in 270 (52.4%) of the 564 children who reported musculoskeletal pain at least once a week in at least one part of the body at baseline. Of the regional pain symptoms, neck pain had highest persistence and, in girls, significantly more than in boys. Persistence of pain was not related to school grade. Widespread pain, determined as in the criteria for fibromyalgia, was found in 132 children (7.5%) and persisted in 35 children (29.7%, 95% CI 21.9 38.4) at follow-up. Disability was more severe in children with pain symptoms in more than one area. This study showed that about half of the preadolescents complaining of musculoskeletal pain at least once a week at baseline had persistent pain symptoms at follow-up. The prognosis of widespread pain in preadolescents was almost the same as the previous findings in adults. PMID- 9414055 TI - Controlled-release oxycodone and morphine in cancer related pain. AB - Controlled-release (CR) formulations of oxycodone and morphine were compared in 45 patients with chronic cancer pain. The study was started with an open-label, randomised titration phase to achieve stable pain control for at least 48 h, followed by a double-blind, randomised, crossover phase in two periods, 3-6 days each. To blind the study using available tablet strengths, the dose ratio of oxycodone to morphine was set at 2:3. A daily telephone contact was maintained between the patient and the investigator. The patients were asked to assess pain intensity four times a day and acceptability of therapy twice daily, and to record possible adverse effects. Pharmacodynamic evaluations were performed at the end of each double-blind period. The patients were allowed to use escape analgesic (respective opioid as oral solution) as needed. Twenty-seven patients were evaluable for both safety and efficacy. Pain was well-controlled during both stable phases. When the period effect was taken into account the two opioids provided comparable analgesia. If the results of the two periods were combined, the patients consumed significantly more escape doses and the mean pain intensities were significantly greater with CR oxycodone. The total opioid consumption ratio of oxycodone to morphine was 2:3 when oxycodone was administered first, and 3:4 when oxycodone was administered after morphine. The total incidence of adverse experiences reported by the patients was similar, but significantly more vomiting occurred with morphine, whereas constipation was more common with oxycodone. PMID- 9414056 TI - A population-based study of the relationship between sexual abuse and back pain: establishing a link. AB - The aim of the present study was to investigate the prevalence of physical and sexual abuse in the general population as well as to investigate the link between abuse and pain. From a pool of randomly selected people 35-45-years-old, three groups were selected based on their reports of their musculoskeletal pain. These were the No Pain Group (n = 449), the Mild Pain Group (n = 229), and the Pronounced Pain Group (n = 271). A group of 142 consecutive patients with chronic musculoskeletal pain was used as a clinical reference group. A standardized questionnaire was employed to determine self-reported physical and sexual abuse. Sexual abuse was more frequently reported than physical abuse and women tended to report more sexual abuse than did men. For women the prevalence of physical abuse ranged from 2% in the No Pain Group to 8% in the Pronounced Pain Group. The total amount of self-reported sexual abuse ranged from 23% in the No Pain Group to 46% in the Pronounced Pain Group. The prevalence of self-reported abuse for the Patient Group differed little from the Pronounced Pain Group and was 35%. For females only, there was a clear link between self-reported abuse and pain as physical abuse increased the risk of pronounced pain by five-fold and sexual abuse increased this risk by four-fold. These data provide the prevalence of self reported abuse in a 'normal' population base and moreover demonstrate an important link between self-reported abuse and pain for women. The findings show that self-reported abuse may be an important predictor for chronic pain and provide support for the idea that abuse may indirectly or directly be implicated in the chronification of pain. PMID- 9414057 TI - A pain education program for chronic cancer pain patients: follow-up results from a randomized controlled trial. AB - The effectiveness of a Pain Education Program in cancer patients with chronic pain offered by nurses was investigated in a randomized controlled clinical trial. A multi-method approach was used in which verbal instruction, written material, an audio cassette tape, and the use of a pain diary were combined to inform and instruct patients about pain and pain management. The Pain Education Program was tailored to the needs of the individual patient and consisted of three elements: (1) educating patients about the basic principles regarding pain and pain management; (2) instructing patients how to report their pain in a pain diary; and (3) instructing patients how to communicate about pain and how to contact health care providers. Following pretesting in 313 patients, patients who needed district nursing and who did not need district nursing at home were randomly assigned to a control or intervention group. Intervention group patients received the Pain Education Program in the hospital, and 3 and 7 days postdischarge by telephone; this was done by nurses who were specially trained as pain counselors. Follow-up assessments were at 2, 4 and 8 weeks postdischarge. Results of the pretest showed that many patients lacked knowledge about pain and pain management. The majority of pain topics had to be discussed. The Pain Education Program proved to be feasible: 75.0% of the patients had read the entire pain brochure, 55.7% had listened to the audio cassette, and 85.6% of pain scores were completed in the pain diary. Results showed a significant increase in pain knowledge in patients who received the Pain Education Program and a significant decrease in pain intensity. However, pain relief was mainly found in the intervention group patients without district nursing. It can be concluded that the tailored Pain Education Program is effective for cancer patients in chronic pain. The use of the Pain Education Program by nurses should be seriously considered on oncology units. PMID- 9414058 TI - Within-person relationships among pain intensity, mood and physical activity in chronic pain: a naturalistic approach. AB - Fifty-seven chronic pain patients rated their pain intensity, mood and activity level, at a random time schedule, eight times a day during 6 consecutive days, according to the Experience Sampling Method (ESM). Within-person correlations among pain intensity, mood and activity level were calculated. We found pain intensity to be significantly associated with mood. However, the associations between pain intensity and activity level, and activity level and mood could not be supported. Further, we examined whether the relationship between pain intensity and mood was the result of a pattern across the day. Results showed that pain intensity and mood were worst in the morning and improved during the afternoon among participants whose pain intensity and mood were correlated significantly. We suggest that attentional as well as behavioural processes might explain the established day pattern of pain intensity and mood. PMID- 9414059 TI - Effectiveness of a multimodal treatment program for chronic low-back pain. AB - In recent years, multidisciplinary pain programs were seen to successfully treat patients by basing treatment on a combination of physical exercise and psychological interventions. However, in spite of their effectiveness, it still remains to be clarified exactly which features of these programs were responsible for patient improvement. Cognitive-behavioral models posit that improvement is due, in part, to changes in patient coping strategies. Nonetheless, as reflected by the conflicting opinions present in the literature, it is questionable whether a so-called 'cognitive shift' is an accurate indicator for return to work of disabled patients. Ninety patients with chronic low back pain took part in a multidisciplinary treatment program. Therapeutic environment reinforces wellness behavior and enhances the patients' sense of control over their pain and resulting disability. The main therapeutic target point was to facilitate return to work. Ways of coping were measured by a well studied coping inventory in the German language (FEKB). Factor analysis revealed three factors: 'catastrophizing', 'search for information' and 'cognitive control'. In addition, assessment included measurements of pain intensity, depression, disability, flexibility of the lumbar spine, and different performance parameters. All of them were measured prior to and at the end of treatment, and following intervals of 6 and 12 months after discharge from program. Measurements showed significant changes over time, but more importantly, nearly all results were seen to stabilize at the 6- and 12-month evaluation following treatment. The coping strategies demonstrated little or poor change. In addition, coping measures and change in coping behavior showed poor prognostic relevance. But other psycho social parameters like self-evaluation of potential return-to-work, application for pension, the length of pre-absence from work, and a decrease in subjective disability following treatment were effective indicators for 'back-to-work'. Other objective parameters, such as medical history, physical impairment and general physical variables were seen to have little predictive value in determining a return to work. The results suggest that the primary target point for further investigation is the analysis of the patients' beliefs about their pain. Our results indicate that future research must be attentive to the complex interactions between environmental factors and the coping demands posed by the specific nature of pain problems. PMID- 9414060 TI - Spinal cord stimulation attenuates augmented dorsal horn release of excitatory amino acids in mononeuropathy via a GABAergic mechanism. AB - Neuropathic pain may be effectively relieved by electric stimulation of the spinal cord (SCS). However, the underlying mechanisms for the ensuing pain relief are poorly understood. In a rat model of neuropathy displaying hypersensitivity to innocuous tactile stimuli, (allodynia), we have earlier demonstrated that SCS may normalise withdrawal response thresholds. In the present study, using microdialysis, it is shown that SCS induces a decreased release of the dorsal horn excitatory amino acids (EAA), glutamate and aspartate, concomitant with an increase of the GABA release. Local perfusion with a GABA(B)-receptor antagonist in the dorsal horn transiently abolishes the SCS-induced suppression of the EAA release. Thus, the effect of SCS on neuropathic pain and allodynia may be due to an activation of local GABAergic mechanisms inhibiting the EAA release which is chronically elevated in such conditions. PMID- 9414061 TI - Mexiletine-induced severe skin eruption, fever, eosinophilia, atypical lymphocytosis, and liver dysfunction. AB - A 64-year-old man developed a severe generalized pruritic morbilliform skin eruption, fever, eosinophilia, atypical lymphocytosis, and liver dysfunction 30 days after ingestion of mexiletine, a sodium channel blocker, prescribed to treat postherpetic neuralgia. Following intravenous dexamethasone, body temperature normalized the next day. However, the skin eruption did not disappear completely for 4 weeks. The patch test was positive for mexiletine. Clinical features and the result of patch test indicated that the patient developed hypersensitivity syndrome, a severe adverse cutaneous drug reaction, caused by mexiletine. We propose that mexiletine be added to the list of drugs that can cause severe adverse cutaneous drug reactions and that patients receiving mexiletine be warned to stop taking the drug immediately if a skin eruption occurs. PMID- 9414062 TI - Pain in multiple leiomyomas alleviated by nifedipine. AB - We have confirmed the usefulness of nifedipine in the treatment of pain present in lesions of multiple skin leiomyomata. Our patient, a 28-year-old woman, had hundreds of skin lesions, proven histologically to be leiomyomata. Nifedipine (10 mg) three or four times daily was remarkably effective in diminishing pain that was more marked in the winter season. PMID- 9414064 TI - Comments on McQuay et al., Pain, 64 (1995) 331-335. PMID- 9414063 TI - Subpial vacuolar myelopathy after intrathecal ketamine: report of a case. AB - Previous studies of intrathecal ketamine use in humans have documented the clinical effects of its administration without reference to local central nervous system (CNS) toxicity (Reich and Silvay, 1989). Although several post-mortem studies in small groups of rabbits, monkeys, and baboons have largely shown no significant CNS damage after intrathecal ketamine use, a study in rats reported vacuolation of dorsal root ganglia (Ahuja, 1983). We report post-mortem CNS histopathological changes of subpial spinal cord vacuolation in a terminally ill cancer patient who received continuous infusion intrathecal ketamine at a rate of 5 mg/day for a duration of 3 weeks. PMID- 9414065 TI - Comments of Yarnitsky et al., Pain, 67 (1996) 327-333. PMID- 9414066 TI - Comments on the use of low-level laser therapy (LLLT) in painful musculo-skeletal disorders. PMID- 9414067 TI - Comments on Zaslansky et al., Pain, 66 (1996) 39-49. PMID- 9414068 TI - Comments on Ripamonti et al., Pain, 70 (1997) 109-115. PMID- 9414069 TI - Comment on Yezierski, Pain, 68 (1996) 185-194. PMID- 9414070 TI - Creatine supplementation as an ergogenic aid for sports performance in highly trained athletes: a critical review. AB - Creatine supplementation has become a common practice among competition athletes participating in different sports over the last few years. The mechanism by which supplementary creatine could have potential ergogenic effects would be an increased muscle creatine and phosphocreatine concentration, leading to a higher rate of ATP resynthesis, a delay in the onset of muscular fatigue and a facilitated recovery during repeated bouts of high-intensity exercise. A critical review of the literature reveals that these ergogenic effects, when found, have been generally shown in untrained subjects performing several exercise bouts under laboratory conditions. The limited body of scientific data available concerning highly trained athletes performing single competition-like exercise tasks indicates that this type of population does not benefit from creatine supplementation. Therefore, the widespread use of creatine ingestion to improve competition performance does not seem to be justified. The potential interest of creatine supplementation for elite athletes could be related to an increased ability to perform repeated high-intensity exercise bouts, either during training or during competition in sports in which repeated efforts are required (e.g. soccer, basketball), but this possibility needs scientific confirmation. PMID- 9414071 TI - Effect of training on antioxidant capacity, tissue damage, and endurance of adult male rats. AB - We studied the effects of physical training on antioxidant defences and susceptibility to damage induced by exhaustive exercise in tissues of adult (12 mo) rats. Therefore, untrained animals were sacrificed either at rest (n = 8) or immediately after swimming to exhaustion (n = 8). Rats trained to swim for 10 weeks were also sacrificed, 48 hr after the last exercise, either at rest (n = 8) or after exhaustive swimming (n = 8). Integrity of mitochondria and sarcoplasmic (SR) or endoplasmic (ER) reticulum of liver, heart, and muscle was assessed by measuring mitochondrial respiratory control and latency of alkaline phosphatase activity. Lipid peroxidation was measured by determination of malondialdehyde and hydroperoxides. Additionally, the effect of training on tissue antioxidant systems was examined by determining the glutathione peroxidase (GPX) and glutathione reductase (GR) activity and the overall antioxidant capacity (CA). Membrane integrity was unaffected by training in liver and muscle, and improved in heart of at rest animals, whereas lipid peroxidation was reduced in both liver and heart. Glutathione peroxidase and glutathione reductase activity, and overall antioxidant capacity were increased (p < 0.05) by training in liver and muscle. In heart, antioxidant capacity was increased from 0.21+/-0.01 to 0.33+/-0.02 (p<0.05), but glutathione peroxidase activity remained unchanged (p>0.05), and glutathione reductase activity was decreased from 3.56+/-0.08 to 2.27+/-0.10 micromol x min(-1) x g(-1) (p < 0.05). The exhaustive exercise gave rise to tissue damage irrespective of trained state, as documented by similar loss of SR and ER integrity, and increase (p<0.05) in lipid peroxidation found in exhausted trained and untrained rats. However, the above changes were elicited by exercise of greater duration in trained than in untrained rats (340+/-17 min and 233+/-6 min, respectively). These findings support the view that free radical-induced damage in muscle could be one of the factors involved in muscle fatigue. If so, the increased endurance in trained rats should reflect lengthening of the time required for the oxidative processes to sufficiently impair cell functions so as to make further exercise impossible. PMID- 9414072 TI - Calcitonin gene-related peptide is increased in hindlimb motoneurons after exercise. AB - Calcitonin gene-related peptide (CGRP) is a neuroactive peptide present in some spinal cord motoneurons and at their motor endplates in skeletal muscle. Although detectable levels of CGRP change after surgical and pharmacological interruptions of neuromuscular connectivity, a clear understanding of its physiological role in the motor system is lacking. The purpose of this study was to investigate whether downhill running exercise, which elicits muscle damage and repair, also elicits changes in CGRP levels in hindlimb motoneurons. Twenty female Wistar rats were divided into five groups: control (c), 48 hours post-exercise (48 hr), 72 hours (72 hr), two weeks (2 wks) and four weeks (4 wks). Exercise groups ran downhill for one 30 minute period. Histological examination of muscle from ankle extensors (triceps surae, TS) and flexors (anterior crural, AC) indicated the characteristic presence of histiocytes by 48 hr post-exercise in TS but not in AC. Paraformaldehyde-fixed, 30 microm sections of lumbar spinal cord (L2-L4) from the same animals were incubated with polyclonal antisera to CGRP. The number of CGRP-positive TS motor nuclei increased significantly by 48 hr after exercise (p = 0.001) vs control and returned to baseline values by 4 wks (p > 0.05). In contrast, no significant changes were observed in the AC motoneuron pool at any post-exercise interval (p > 0.05). The temporal changes in CGRP levels in TS motoneurons suggest that expression of this neuropeptide may be differentially regulated by exercise-induced changes in neuromuscular function, possibly as related to muscle tissue damage/repair mechanisms and thus to remodeling at the neuromuscular junction. PMID- 9414073 TI - Characteristics of body composition and muscle strength in college Sumo wrestlers. AB - The purpose of this study was to investigate the characteristics of body composition and force generation capacity in college Sumo wrestlers (N=13, age=19.8+/-0.3 yr, stature= 178.5+/-1.6 cm, body mass = 111.2+/-3.8 kg, X+/-SE) in comparison with untrained males (N=18, 20.3+/-0.2 yr, 170.1+/-1.7 cm, 59.2+/ 1.4 kg). The Sumo wrestlers had significantly higher average values in relative fat mass (24.8+/-1.0%) and fat-free mass (83.3+/-2.0 kg), estimated by an underwater weighing method, than the untrained subjects (relative fat mass = 12.9+/-0.1 %, fat-free mass = 51.5+/-1.3 kg). Moreover, the Sumo wrestlers had 1.7 to 1.9 times greater cross-sectional areas (CSAs) of elbow flexors and extensors and knee extensors, determined by a B-mode ultrasound technique, compared to those of the untrained subjects. Force values produced during elbow flexion and extension and knee extensions tasks under isokinetic contraction mode at constant velocities of 1.05, 3.14 and 5.24 rad x s(-1) were significantly higher in the Sumo wrestlers than in the untrained subjects. The force value in all test conditions was significantly correlated to the related-muscle CSA, r=0.611-0.910 (p<0.05). The difference between the two groups in force per unit CSA (F x CSA[-1]) during elbow flexion was not significant at all test velocities. However, the Sumo wrestlers showed significantly lower F x CSA(-1) values in elbow extension at 5.24 rad x s(-1) and in knee extension at all test velocities. Thus, the body composition of the Sumo wrestlers was characterized by a high fat content and a large fat-free mass. Moreover, the Sumo wrestlers had considerably larger muscle CSAs of limbs than the untrained subjects. For the Sumo wrestlers, however, force output of the muscles with a pennate structure were not proportional to their CSAs, particularly in knee extensors. PMID- 9414074 TI - Changes in upper body power following heavy-resistance strength training in college men. AB - The purpose of this study was to determine the effects of heavy-resistance strength training on measures of bench press power (BPP) using absolute loads and seated shot put (SSP) performance. Twenty-four college men were measured for 1-RM bench press, BPP, and SSP before and after weight training twice weekly for 12 weeks. BPP was measured with free weights using a digital timing system and randomly assigned loads equivalent to 30%, 40%, 50%, 60%, 70% and 80% of the 1 RM. Post-training tests used the same absolute loads as during the pre-training test to assess BPP. Following training BPP increased significantly at each load, shifting the power curve upward by an average of 13.6%. The 1-RM bench press increased significantly by 9.1%, but the SSP increased nonsignificantly by only 1.8%. Peak power was produced at approximately 40-50% of the 1-RM before and after training. Changes in SSP distance were nonsignificantly correlated (r=0.27 0.20) with the increases in BPP. Resistance training shifts the power curve in a positive direction when the measurements are determined with absolute loads, but the increased power may not be transferred to an absolute performance task like the SSP. PMID- 9414075 TI - Isokinetic muscle strength and capacity for muscular knee joint stabilization in elite sailors. AB - In the present study isokinetic dynamometry was used to evaluate the capacity for dynamic knee joint stabilization via muscle contraction in elite sailors (15 males, SM; 6 females, SF) compared to a group of matched controls (8 males, CM). Maximal concentric, eccentric and isometric moment of force (peak moment and moment at 50 degree knee flexion) was obtained for the knee extensors (quadriceps) and flexors (hamstrings) during isokinetic knee joint movement at angular velocities 0, 30, 120 and 180 degrees x s(-1). High levels of eccentric knee extension strength were observed for the elite sailors compared to the controls (p < 0.05). Based on peak moment and 50 degree moment, respectively, conventional hamstring/quadriceps (H/Q) strength ratio (+/-SD) ranged from 0.37+/ 0.06 to 0.54+/-0.06 and from 0.42+/-0.07 to 0.57+/-0.10 across groups, speed and contraction mode. The female elite sailors displayed lower (p<0.05) concentric H/Q ratios at 120 and 180 degrees x s(-1) compared to the controls (0.41-0.45 vs. 0.51-0.56, respectively). The ratio of eccentric hamstring to concentric quadriceps strength (H/Q for extension) or concentric hamstring to eccentric quadriceps strength (H/Q for flexion) may provide a more functional estimate of the capacity for muscular knee joint stabilization (1). Based on peak moment and 50 degree moment, respectively, this "functional" H/Q ratio ranged from 0.24+/ 0.03 and 0.25+/-0.02 for knee flexion at 180 degrees x s(-1) to 0.97+/-0.17 and 0.88+/-0.12 for knee extension at 180 degrees x s(-1) among the three groups. Comparable levels of "functional" H/Q ratio were observed (p>0.05) for fast knee extension in the elite sailors (SF:0.81-0.97, SM: 0.88-0.95) and the male controls (CM: 0.80-0.84). In conclusion, a "functional" H/Q ratio of 0.8-1.0 observed for all subjects indicated a significant functional capacity of the hamstring muscles for providing muscular stability at the knee joint in fast knee extension. A significant potential for muscular knee joint stabilization appeared for the elite sailors despite their high maximal quadriceps strength and partially lower (SF) conventional H/Q ratios. PMID- 9414076 TI - The plasma lactate response to exercise and endurance performance: relationships in elite triathletes. AB - The lactate response to exercise has been studied thoroughly during the last decades and it has been described using a variety of terms and definitions. Numerous investigations observed close relationships between the lactate response and endurance performance. The main question in this study was which of the various lactate responses during incremental exercise described in the literature was the best indicator of endurance performance. The plasma lactate response (PLR) was assessed during an incremental exercise test on 13 male elite triathletes (age 25.5+/-5.8 yrs; HT 179.7+/-5.4 cm; WT 71.3+/-4.7 kg) on a bicycle ergometer. The load was started at 2.5 W/kg and increased by 40 W every 4 min. We evaluated the following PLR-parameters: the workloads at the fixed lactate levels of 2, 3, 4, 5, 6, 7, and 8 mmol/l which were assessed by extrapolation from a workload-lactate-heart rate curve (P2, P3, P4, P5, P6, P7, P8 respectively), the lactate threshold which was defined as the workload at the point at which a non-linear increase of blood lactate occurred (Plt), and the workload at the lactate level that was 1 mmol/l above the baseline (P + 1). Four to seven weeks after the laboratory test, heart rate and lactate levels were assessed during a 40-km long time trial on a bicycle. Two parameters were considered as indicative of athletic performance: the road racing time (Tt), and the workload extrapolated from the workload-lactate-heart rate curve at the heart rate and lactate levels observed during the time trial (Pt). Only P2 showed a significant correlation with Tt (r=-0.65; p < 0.05; se = 72.5 s). Multiple regression analysis with the anthropometric parameters height and weight as additional independent parameters did not change the predictive value. We concluded that for predicting the cycling performance of similarly well-trained subjects the predictive value of PLR is negligible. PMID- 9414077 TI - Validity of near-infrared interactance for estimating relative body fat in female high school gymnasts. AB - The present investigation examined the validity of near-infrared interactance (NIR) estimates of relative body fat (% fat) from the Futrex-5000 (F5000), Futrex 5000A (F5000 A), and Futrex-1000 (F1000) instruments. Ninety-eight female high school gymnasts (X age+/-SD = 15.7+/-1.2 yr) participated in this investigation. Subsamples were used to cross-validate the F5000 (n = 52), F5000A (n = 46), and F1000 (n = 89) instruments. The NIR % fat estimates were validated against criterion % fat from underwater weighing (UWW) using the adult conversion constants of Brozek et al. (6) (UWWB) and the female age-specific constants of Lohman (23) (UWWL). The cross-validation statistical analysis included examination of the constant error (CE), standard error of estimate (SEE), r, and total error (TE). In addition, full-model multiple regression analyses were used to predict UWWB or UWWL from body weight (BW) and height (HT). BW and HT were correlated with % fat at R = 0.65-0.70, while the validity coefficients for the NIR instruments ranged from r = 0.40-0.78. The F5000 resulted in nonsignificant CE values (-0.3 % fat vs UWWB and 1.5 % fat vs UWWL; p > 0.008) as well as the lowest TE values (TE = 3.1 % fat vs UWWB and 4.0 % fat vs UWWL). All other NIR estimates of % fat resulted in TE values > or = 6.3 % fat. In addition, for all NIR instruments there were negative correlations for the plots of the CE versus the mean of predicted and criterion (UWWB and UWWL) % fat. Therefore, the present findings indicated that the F5000 provided more accurate estimates of % fat than the F5000A or F1000 instruments, but may underestimate the desired minimal body weight for female gymnasts at the low end of the % fat distribution. PMID- 9414078 TI - Circadian rhythms have no effect on cycling performance. AB - The aim of this research was to determine if circadian rhythms have an effect on time trial cycling performance of 15 min duration. Seven males (Mean+/-SD): age, 22.3+/-4.9 yr; height 179.0+/-7.9 cm, body mass 74.5+/-15.5 kg; VO2max 68.0+/-5.7 ml x kg(-1) x min(-1) who were all competitive cyclists or triathletes with previous experience in laboratory testing procedures volunteered to participate in this study. Each of the seven subjects underwent a series of four tests; one VO2 max test, and three 15 min maximal performance tests, at varying times during a 24 hr period. Testing times were at 08.00-10.00; 14.00-16.00 and 20.00-22.00 hours. Heart rate was recorded during the last 10-15 seconds of each minute and blood lactate levels were taken at 5 and 10 min during exercise and again immediately post-exercise. O2 consumption was measured continuously using open circuit spirometry. RPE was measured using the Borg scale at 5 and 10 min during, and again immediately following the completion of testing. Resting oral temperature was the only variable to show a significant time of day effect (p<0.05). Oral temperature during the afternoon was higher than both morning and evening results by 0.76 degrees C and 0.09 degrees C respectively. Total work (kJ) and average power output (W) were recorded at their highest during the morning session and reached a trough during the afternoon session, but these differences were not significant (p = 0.9997 and 0.9972 respectively). The results obtained in this study indicate that while certain biological rhythms are present, they appear to have no effect on this type of cycling performance. Although athletic performance may be enhanced by training programs that are compatible with an individuals body clock, the ability to perform and train at various times has an adaptive response which appears to over-ride these naturally inherent rhythms. PMID- 9414079 TI - Pre-exercise carbohydrate meal and endurance running capacity when carbohydrates are ingested during exercise. AB - This study examined whether combining a pre-exercise carbohydrate meal with the ingestion of a carbohydrate-electrolyte solution during exercise is better in improving endurance running capacity than a carbohydrate-electrolyte solution alone. Ten men completed three treadmill runs at 70% VO2max to exhaustion. They consumed 1.) a carbohydrate meal three hours before exercise and a carbohydrate electrolyte solution during exercise (M + C), or 2.) a liquid placebo three hours before exercise and the carbohydrate-electrolyte solution during exercise (P + C), or 3.) a placebo three hours before exercise and placebo during exercise (P + P). When the meal was consumed (M + C) serum insulin concentrations were higher at the start of exercise, and carbohydrate oxidation rates were higher during the first 60 min of exercise compared with the values found in the P + C and P + P trials (p < 0.01). Exercise time was longer in the M + C (147.4+/-9.6 min) compared with the P + C (125.3+/-7 min) (p < 0.01). Also, exercise time was longer in M + C and P + C compared with the P + P (115.1+/-7.6 min) (p < 0.01 and p < 0.05 respectively). These results indicate that the combination of a pre exercise carbohydrate meal and a carbohydrate-electrolyte solution further improves endurance running capacity than the carbohydrate-electrolyte solution alone. PMID- 9414080 TI - Familial aggregation of leisure-time physical activity -- a three generation study. AB - Studies of parental influence on children's physical activity have had different results. Parental effect on physical activity during adolescence is less studied, and three generation studies have not been carried out. The purpose of our study was to examine intra- and intergenerational associations of leisure time physical activity among family members in three generations. Due to the major changes in society during this time, we also took into consideration the socioeconomic status of the adult subjects. The material consisted of 3254 twins at the age of 16, their parents and grandparents. Twins and their parents received a questionnaire in 1991-1993, which included questions about the health and lifestyle, socioeconomic status and leisure time physical activity. The parents' questionnaire also included questions about their own parents' leisure time physical activity and socioeconomic status. Based on these questions adolescents, parents and grandparents were classified into physical activity classes. The socioeconomic classification of parents and grandparents was based on their occupation. Intragenerational physical activity patterns were significantly associated with each other. Among adolescents the strongest correlation were between monozygotic boys (0.720) and monozygotic girls (0.638). Physical activity patterns were not associated between generations, but there was a significant difference between very active and inactive mothers concerning their daughters' physical activity. Farmers had the lowest proportion of very active subjects only among the parental generation. Because physical activity patterns do not appear to be transmitted from one generation to the next, it is probable that by constant training and education we can obtain the benefits of physical activity. PMID- 9414081 TI - Indications of prevalence, practice and effects of anabolic steroid use in Great Britain. AB - A growing number of reports of anabolic-androgenic streroid (AS) use in Great Britain (GB) among non-competitive groups have emerged since the beginning of 1990s. A study was commissioned by the Departments of Health for England, Scotland and Wales, to explore the extent and uses of AS from the public health point of view. As a part of a wider investigation into AS use, 21 gymnasia in England, Scotland and Wales were surveyed by questionnaire. The response rate was 59%. We found that of the 1667 participants, 9.1% of the men and 2.3% of the women had taken AS at some time and 6% of the men and 1.4% of the women were current users. Considerable variation in the prevalence of use was found, ranging from no reports in three of the gymnasia, up to 46%. We also investigated patterns of AS use and perceived side-effects in a wide-ranging group of AS users (n = 110), who were recruited through social networks. In-depth interviews with the users revealed that the 97 men (27+/-7 years) and 13 women (25+/-5 years) had been using AS regularly for 2.05+/-1.7 years and 1.9+/-2 years, respectively. Seventy-two injected AS. While most injected themselves, 25% were mainly injected by their friend. Up to 16 different drugs were taken by interviewees during the present or last cycle. Polydrug use was common and dosage taken exceeded therapeutic recommendations. Sixteen interviewees did not report side-effects, while the majority reported two or more. Many of these were cosmetic. Of the 97 men interviewed, 56% reported testicular atrophy, 52% gynaecomastia, 36% elevated blood pressure, 56% fluid retention, 26% injuries to tendons, 22% nosebleeds and 16% more frequent colds. Six men reported problems with kidney function and five with liver function. Problems with sleep were reported by 37%. Of the 13 women interviewed, eight reported menstrual irregularities, eight fluid retention, four clitoral enlargement, three decreased breast size and two elevated blood pressure. Four reported sleeplessness. PMID- 9414082 TI - Botulinum neurotoxin types A and E require the SNARE motif in SNAP-25 for proteolysis. AB - Botulinum neurotoxins type A and E (BoNT/A and BoNT/E) are metalloproteases with a unique specificity for SNAP-25 (synaptosome-associated protein of 25 kDa), an essential protein component of the neuroexocytotic machinery. It has been suggested that this specificity is directed through the recognition of a nine residue sequence, termed SNARE motif, that is common to the other two SNARE proteins: VAMP (vesicle-associated membrane protein) and syntaxin, the only known substrates of the other six clostridial neurotoxins. Here we analyse the involvement of the four copies of the SNARE motif present in SNAP-25 in its interaction with BoNT/A and BoNT/E by following the kinetics of proteolysis of SNAP-25 mutants deleted of SNARE motifs. We show that a single copy of the motif is sufficient for BoNT/A and BoNT/E to recognise SNAP-25. While the copy of the motif proximal to the cleavage site is clearly involved in recognition, in its absence, other more distant copies of the motif are able to support proteolysis. Also, a non-neuronal isoform of SNAP-25, Syndet, is shown to be sensitive to BoNT/E, but not BoNT/A, whilst the SNAP-25 isoforms from Torpedo marmorata and Drosophila melanogaster were demonstrated not to be substrates of these metalloproteases. PMID- 9414083 TI - Hyaluronectin blocks the stimulatory effect of hyaluronan-derived fragments on endothelial cells during angiogenesis in vitro. AB - Hyaluronic acid (HA) is a glycosaminoglycan of the extracellular matrix. Its fragmentation by the hyaluronidase, secreted by tumor cells, facilitates tumor invasion and the HA degradation products generated stimulate angiogenesis. We report here that the HA-binding protein hyaluronectin (HN) inhibits the stimulatory effect of HA-derived fragments on the proliferation and migration of endothelial cells in vitro, and hampers the organization of endothelial cells into capillary-like structures. Since HN strongly inhibits endothelial cell adhesion to immobilized HA, it is postulated that HN acts by impairing the binding to endothelial cells of HA fragments generated by hyaluronidase, thereby neutralizing the effect of HA degradation products on angiogenesis. Our results reveal a new mechanism by which the angiogenesis induced by HA fragments is modulated by HN. PMID- 9414084 TI - Kinetic mechanism of active site non-equivalence in transketolase. AB - The two-step mechanism of coenzyme (TDP) binding to apotransketolase has been examined by kinetic modeling, and the rate and equilibrium constants for each binding step for two active sites have been determined. The dissociation constants for the primary fast binding step and the forward rate constants for the secondary slow binding step have been shown to be similar for two active sites. The backward rate constants for the secondary binding step are different for two active sites, providing the kinetic mechanism of their non-equivalence in TDP binding. PMID- 9414085 TI - Protective effects of the lipophilic redox conjugate tocopheryl succinyl-ethyl ferulate on HIV replication. AB - Previously, we demonstrated that ferulate ethyl and tocopherol reduced HIV replication. In this study, we investigate whether the conjugation of both compounds (O-tocopheryl succinyl O-ethyl ferulate) can increase HIV inhibition. We show here for the first time that O-tocopheryl succinyl O-ethyl ferulate inhibits 80% of HIV replication (HIV-1 acute infection and HIV transmission), inhibits cell lipoperoxidation and prevents cellular glutathione consumption. Compared to ferulate ethyl and tocopheryl succinyl, O-tocopheryl succinyl O-ethyl ferulate inhibits more HIV replication. This may be due in part to the great increase in the lipophilicity of this compound. PMID- 9414086 TI - Upregulation of MMP-9 expression in MDA-MB231 tumor cells by platelet granular membrane. AB - The interaction between tumor cells and platelets facilitates the formation of metastasis in a way depending on the platelet aggregating ability of the tumor cell, but the mechanism remains to be elucidated. We have shown, by zymography and Western blot, that platelets greatly increased the secretion to the culture medium of MMP-9 by human mammary tumor cells MDA-MB231. This increase, which was dependent on protein synthesis, was caused by the platelet aggregates interacting with the tumor cells and not by the soluble factors released during platelet activation. Platelet subcellular fractionation allowed the localization of the inducing factor to the membrane fraction of the platelet granules, thus requiring platelet aggregation in order to become accessible on the platelet surface. PMID- 9414087 TI - Identification of a novel nuclear speckle-type protein, SPOP. AB - A novel antigen recognized by serum from a scleroderma patient was identified by expression cloning from the HeLa cell cDNA library. The cloned cDNA encoded a 374 amino acid protein with a relative molecular mass of 47,000 and a predicted amino acid sequence 62.7% identical to the hypothetical protein of Caenorhabditis elegans, T16H12.5. The deduced amino acid sequence had a typical POZ domain and an unidentified region conserved during evolution. No zinc finger or RNA recognition motifs were found in this clone. The 2 kbp mRNA encoding the novel clone SPOP (speckle-type POZ protein) was found to be expressed in all human tissues examined. HA-tagged SPOP, transfected and overexpressed in COS7 cells, exhibited a discrete speckled pattern in the nuclei and was co-localized with the splicing factor, snRNP B'/B. Deletion analysis revealed that both the POZ domain and the evolutionarily conserved region at the amino-terminus are required for the nuclear speckled accumulation of SPOP. PMID- 9414088 TI - Ribosomal efficiency and growth rates of freshly isolated Escherichia coli strains originating from the gastrointestinal tract. AB - It has been previously reported that for natural Escherichia coli isolates from the ECOR collection, there were differences in the ribosomal efficiencies and there was a direct correlation between growth rate and the ribosome efficiency (R factor). The aim of this study was to determine whether strains freshly isolated (i.e. subcultured < 5 times) from the gastrointestinal tract ecosystem also exhibited this correlation. Eleven E. coli and two Enterobacter spp. isolates from either humans, pigs, rats or a mammoth were investigated. Considerable variability in the R-factor was noted using an in vitro translation assay, however no consistent correlation between the R-factor and growth rate was noted. PMID- 9414090 TI - The 'assembly-promoting sequence region' of microtubule-associated protein 4 failed to promote microtubule assembly. AB - In order to study the function of the bovine MAP4 microtubule-binding domain (the assembly-promoting (AP) sequence region), a fragment corresponding to the AP sequence region was prepared using an Escherichia coli expression system. When the fragment was mixed with purified tubulin at 37 degrees C, the fragment caused a time- and dose-dependent turbidity increase, and the fragment bound to tubulin. However, the products were cold-stable, and amorphous aggregates were observed by electron microscopy. Using axonemes as the seeds for microtubule assembly, the microtubule-elongating activity of the fragment was examined. A dose-dependent turbidity increase of the sample was observed, and electron microscopic observation revealed that microtubules were dose-dependently elongated from the axonemes. Consequently, the AP sequence region does not nucleate microtubules, but elongates them. PMID- 9414089 TI - The promyelocytic leukemia protein PML has a pro-apoptotic activity mediated through its RING domain. AB - The promyelocytic leukemia protein PML is known to form nuclear multiprotein complexes which are compromised in several pathogenic conditions including acute promyelocytic leukemia. We show that in cells infected with a single stranded RNA virus, which relocates PML bodies to the cytoplasm, the infected cells are more resistant to serum starvation induced apoptosis than their uninfected counterparts. Antisense PML oligonucleotides increase cell survival under serum deprivation conditions indicating that PML is directly involved in the apoptotic activity. Transient transfection studies have indicated that this pro-apoptotic activity of PML is mediated through the zinc binding region known as the RING finger. Viral attack of PML nuclear bodies appears to allow the virus to deregulate host cell apoptotic machinery in order to establish chronic infection. PMID- 9414091 TI - Tricyclic antidepressants block cholinergic nicotinic receptors and ATP secretion in bovine chromaffin cells. AB - Nicotine-induced ATP secretion from chromaffin cells was blocked by imipramine and desipramine. This blocking action took place on both, fast and slow, components of ATP secretion. Exposure of chromaffin cells to nicotine (10 microM) for 4 s induced an inward current of about -155 pA. Imipramine and desipramine blocked, in a concentration-dependent manner, both peak inward current and total charge influx in response to nicotine. In addition, imipramine and desipramine partially (40%) blocked depolarization-induced ATP secretion and Ca2+ currents evoked by high K+. This suggests that tricyclic antidepressants block nicotine induced ATP secretion by acting on two targets: one is the nicotinic receptor itself and the second one are voltage-dependent Ca2+ channels. PMID- 9414092 TI - Representation of amino acid sequences in terms of interaction energy in protein globules. AB - We suggest a new simple approach for comparing the primary structure of proteins and their spatial structure. It relies on the one-to-one correspondence between each residue of the polypeptide chain and the energy of van der Waals interactions between the regions of the native globule flanking this residue. The method obviates the sophisticated geometrical criteria for estimating similarity between spatial structures. Besides, it permits one to analyze structural units of different scale. PMID- 9414093 TI - Comparison of control analysis data using different approaches: modelling and experiments with muscle extract. AB - Experimental and model studies have been performed to characterize the control properties of hexokinase and phosphofructokinase in muscle glycolysis and to examine the nature of error associated with experimental flux control coefficient determinations. Different approaches of metabolic control analysis, classical titration, co-response analysis and kinetic modelling indicated that flux control coefficients could be reliably estimated experimentally for the upper part of glycolysis. The kinetic parameters applied to construct the mathematical model were determined in muscle extract under similar conditions used for flux studies. If the kinetic parameters of commercial enzymes are introduced into the model the control analysis data cannot be trusted. Co-response analysis can also be successfully applied to determination of the flux control coefficients of the system. However, the involvement of a rapid-equilibrium enzyme, such as glucose 6 phosphate isomerase, could result in estimation errors for the relevant co response coefficients that are propagated into the elasticity matrix. If the co response coefficients related to isomerase activity are replaced by the values obtained by kinetic modelling, the values of elasticities are correct. Our data also suggest that in the upper part of glycolysis hexokinase mainly controls the pathway flux whereas phosphofructokinase exerts dominant control on the turnover of internal metabolite stocks inside the system. PMID- 9414094 TI - The in vitro DNA binding properties of NDP kinase are related to its oligomeric state. AB - Genetic and biochemical evidences suggest that the enzymatic activity of NDP kinase is necessary but not sufficient for its biological function. While the human NDPK-B binds specifically single-strand polypyrimidines sequences, the hexameric enzyme from Dictyostelium does not. We demonstrated by electrophoretic mobility shift assay and filter binding assay that a dimeric mutant from Dictyostelium binds to an oligodesoxynucleotide while the wild-type does not. These data suggest that the differences in the DNA binding properties of several eucaryotic NDP kinases might be correlated to the differences in the stability of their hexameric structure. PMID- 9414095 TI - Enhancement of fibrin binding and activation of plasminogen by staplabin through induction of a conformational change in plasminogen. AB - Staplabin (0.3-0.6 mM), a fungal triprenyl phenol, enhanced 3-fold the plasminogen activator-catalyzed activation of Glu-plasminogen and Lys-plasminogen as well as their binding to fibrin. Staplabin was not stimulatory to the amidolytic activity of plasmin and plasminogen activators. Even in the presence of epsilon-aminocaproic acid (EACA) and fibrinogen fragments, allosteric effectors for Glu-plasminogen, staplabin increased the activation of both forms of plasminogen. In size-exclusion chromatography of Glu-plasminogen and Lys plasminogen, the molecular elution time, which varies as the conformation of a protein changes, was shortened by staplabin. These results suggest that staplabin causes plasminogens to be more susceptible to activation and fibrin binding by inducing a conformational change that is, at least in part, different from that induced by EACA and fibrinogen fragments. PMID- 9414096 TI - Modulation of fructose-2,6-bisphosphate metabolism by components of the extracellular matrix in cultured cells. Interaction with epidermal growth factor. AB - The use of NIH3T3 fibroblasts overexpressing different mutations of the EGF receptor shows that regulation of fructose-2,6-bisphosphate (Fru-2,6-P2) metabolism by EGF is mediated by the kinase activity of the EGF receptor and suggests a PLCgamma1-mediated mechanism. The effect of several extracellular matrix components on glucose metabolism was assessed by incubating A431 cells and NIH3T3 fibroblasts with heparin, laminin, fibronectin, collagen and PG-I and PG II proteoglycans and measuring the levels of Fru-2,6-P2. Laminin increased the levels of Fru-2,6-P2 and heparin decreased the levels of the metabolite, whereas the other molecules did not have any effect. No effect of laminin or heparin in glucose uptake by the cell was observed. Laminin was able to modulate the effects of EGF on Fru-2,6-P2 concentration, suggesting cross-talk between these agents. PMID- 9414097 TI - A RIPE3b1-like factor binds to a novel site in the human insulin promoter in a redox-dependent manner. AB - In the human insulin gene, a regulatory sequence upstream of the transcription start site at -229 to -258 (the E2 element) binds a ubiquitous factor USF. The present study led to the identification of a second factor, D0, that binds to an adjacent upstream site, the C2 element, that has previously not been described. The results demonstrate that D0 exhibits similar properties to RIPE3b1, a factor shown to be an important determinant of insulin gene beta-cell-specific expression. Binding of D0 to the C2 element was abolished by the oxidising agent diamide, and the alkylating agent N-ethylmaleimide. The results indicate that expression of the insulin gene may be regulated by a redox-dependent pathway involving RIPE3b1 or a RIPE3b1-like factor. PMID- 9414098 TI - Cytotoxicity of spin trapping compounds. AB - Spin trapping compounds are used frequently to detect free radicals released by cells. Their cytotoxicity has to be considered in order to prevent perturbations of normal cell growth and viability. Eleven spin traps (eight nitrones and three nitroso traps) have been tested for their effects on bovine aortic endothelial cells (toxicity range, 50% survival rate). The lowest cytotoxicity was found for 5,5-dimethylpyrroline-1-oxide and 2,2,4-trimethyl-2H-imidazole-1-oxide whereas nitrosobenzene and 2-methyl-2-nitrosopropane exerted the strongest cytotoxic effects. In addition, three nitronyl nitroxides were tested. Their cytotoxicity was found to be dependent on substitution, and the toxic concentration of a lipophilic derivative was found to be more than two orders lower as compared to a hydrophilic derivative. The results of this study indicate that most spin traps can be used in cell cultures at customary (i.e. millimolar) concentrations; caution is recommended when nitroso spin traps are applied to cells. PMID- 9414099 TI - GC rich DNA oligonucleotides with narrow minor groove width. AB - Investigation of the width of the minor groove using 500 MHz NMR spectroscopy in three closely related 11-mer B-DNA duplexes shows that the minor groove is narrow in a GC rich oligonucleotide, and that a narrow minor groove is not something endemic to DNAs with persistent repetitions of adenine nucleotides (A-tract DNA). The width of the groove is dictated by local sequence contexts and independent of neighboring A-tract DNA. PMID- 9414100 TI - Differential internalisation of mGluR1 splice variants in response to agonist and phorbol esters in permanently transfected BHK cells. AB - The internalisation of metabotropic glutamate receptor (mGluR1alpha) and its splice variant (mGluR1beta), in response to agonist and phorbol esters (PMA), has been studied. Both mGluR1alpha and mGluR1beta exhibit a similar rate of internalisation following PMA treatment, with a shift in their distribution from plasma membrane to endosome-enriched membrane fractions. Agonist challenge however caused a rapid loss, within 5-10 min, of mGluR1beta but not mGluR1alpha from the cell surface. These results show that the two forms of mGluR1 show different internalisation responses to agonist and suggest that the C-terminal region of the molecule plays an important role in this phenomenon. PMID- 9414101 TI - Chemical oxidation and DNA damage catalysed by inorganic sunscreen ingredients. AB - Titanium dioxide (TiO2) has been noted (US Federal Register, 43FR38206, 25 August 1978) to be a safe physical sunscreen because it reflects and scatters UVB and UVA in sunlight. However, TiO2 absorbs about 70% of incident UV, and in aqueous environments this leads to the generation of hydroxyl radicals which can initiate oxidations. Using chemical methods, we show that all sunscreen TiO2 samples tested catalyse the photo-oxidation of a representative organic substrate (phenol). We also show that sunlight-illuminated TiO2 catalyses DNA damage both in vitro and in human cells. These results may be relevant to the overall effects of sunscreens. PMID- 9414102 TI - Comparative mechanisms and rates of free radical scavenging by carotenoid antioxidants. AB - The comparative mechanisms and relative rates of nitrogen dioxide (NO2.), thiyl (RS.) and sulphonyl (RSO2.) radical scavenging by the carotenoid antioxidants lycopene, lutein, zeaxanthin, astaxanthin and canthaxanthin have been determined by pulse radiolysis. All the carotenoids under study react with the NO2. radical via electron transfer to generate the carotenoid radical cation (Car.+). In marked contrast the glutathione and 2-mercaptoethanol thiyl radicals react via a radical addition process to generate carotenoid-thiyl radical adducts [RS-Car].. The RSO2. radical undergoes both radical addition, [RSO2-Car]. and electron abstraction, Car.+. Both carotenoid adduct radicals and radical cations decay bimolecularly. Absolute rate constants for radical scavenging were in the order of approximately 10(7)-10(9) M(-1) s(-1) and follow the sequence HO(CH2)2S. > RSO2. > GS. > NO2.. Although there were some discernible trends in carotenoid reactivity for individual radicals, rate constants varied by no greater than a factor of 2.5. The mechanism and rate of scavenging is strongly dependent on the nature of the oxidising radical species but much less dependent on the carotenoid structure. PMID- 9414103 TI - Human monocyte-derived and CD34+ cell-derived dendritic cells express functional receptors for platelet activating factor. AB - Dendritic cells (DC) are a heterogeneous population of specialized antigen presenting cells that exhibit complex trafficking properties. DC differentiated in vitro from both peripheral monocytes and CD34+ cells expressed mRNA for platelet activating factor (PAF) receptor. Expression of PAF receptor was increased by TNF alpha, a prototypic inflammatory cytokine that induces differentiation and in vivo mobilization of DC. PAF induced in vitro directional migration of DC obtained from both precursor cells through its specific receptor. Since DC are highly motile cells, protein chemoattractants as well as bioactive phospholipids are likely to contribute to tissue localization of DC, in vivo. PMID- 9414105 TI - The rate-limiting steps for the folding of an antibody scFv fragment. AB - The refolding kinetics of a single-chain Fv (scFv) fragment, derived from the phosphorylcholine binding antibody McPC603, was investigated. Both prolyl-peptide bonds which are cis in the native state affect the refolding kinetics of long term denatured protein. The rate-limiting step is the trans --> cis isomerization at the ProL95-peptide bond, which is catalyzed by peptidyl-prolyl-cis/trans isomerase (PPIase), and is the prerequisite for correct V(H)/V(L) domain association. Refolding of short-term denatured protein resulted in complex refolding kinetics, too. This kinetic heterogeneity could be ascribed to cis --> trans re-isomerization at the ProL95-peptide bond to the wrong conformation in a folding intermediate. PPIase was shown to increase the fraction of slowly folding species, thereby competing with the fast folding of short-term denatured scFv, having native proline conformations. A trapped intermediate is rapidly populated, and the return from this state becomes rate-limiting. PMID- 9414104 TI - Cell cycle dependent toxicity of an amphiphilic synthetic peptide. AB - The cytotoxic properties of an amphiphilic synthetic peptide are presented. Comparative analysis of proliferating, differentiated and confluent H9C2 adherent cells and L1210 cells in suspension shows a correlation between toxicity and cell stage (proliferating cells). Electrophysiological measurements on Xenopus laevis oocytes bathed in the peptide also demonstrated the induction of cationic currents, which is voltage and phosphate dependent. These results allow us to hypothesize that the observed toxicity is related to membrane hyperpolarization of proliferating cells at the G1/S cell cycle phase transition. PMID- 9414106 TI - Characterisation of the rat and mouse homologues of gC1qBP, a 33 kDa glycoprotein that binds to the globular 'heads' of C1q. AB - gC1qBP is a 33 kDa glycoprotein that binds to the globular 'heads' of C1q. We have cloned cDNAs encoding the rat and mouse homologues of gC1qBP. Comparison of the cDNA-derived amino acid sequences of gC1qBP reveals that either of the rodent sequences is 89.9% identical to the reported human sequence. Recombinant rat gC1qBP binds avidly to human C1q. gC1qBP mRNA is abundantly expressed in every rat and mouse tissue analysed. Rat mesangial cells synthesise gC1qBP, but do not express gC1qBP on the cell surface. In rat serum, gC1qBP is present at low levels. PMID- 9414107 TI - Promoters of epithelialization induce expression of vascular endothelial growth factor in human gastric epithelial cells in primary culture. AB - Both epithelialization and angiogenesis are indispensable processes in gastric ulcer healing. Coordination between these processes has not been well studied. In the present study, we have established a new primary culture system of human gastric epithelial cells and investigated the effect of epithelialization stimulants on a specific angiogenic factor, vascular endothelial growth characterized as epithelial cells. Both epithelialization stimulants, hepatocyte growth factor (HGF) and epidermal growth factor (EGF), significantly stimulated vascular EGF expression in gastric epithelial cells. HGF and EGF receptors were expressed by the cells, suggesting that regulation may be mediated through specific receptors. PMID- 9414108 TI - Protein kinase C (PKC) epsilon enhances the inhibitory effect of TNF alpha on insulin signaling in HEK293 cells. AB - Recently we have shown that PKC beta1 and beta2 are able to inhibit the tyrosine kinase activity of the human insulin receptor (HIR). Now we have investigated whether a distinct PKC isoform might be involved in the inhibitory effect of TNF alpha on insulin signaling in HEK293 cells. TNF alpha induces a rapid translocation of the PKC isoform epsilon (TNF alpha 10(-9) M, maximal effect within 5-10 min) in rat-1 fibroblasts, while no effect occurred on other isoforms. Cotransfection of HIR with PKC epsilon did not significantly reduce the insulin stimulated receptor kinase activity; however, when cells were incubated with TNF alpha for 10 min (10(-9) M) a 62 +/- 17% (n = 5) inhibition of the insulin receptor kinase activity was observed which was significantly (P<0.01) higher than that observed in cells which were not transfected with PKC (32 +/- 11.5%, n = 5). The data suggest that translocation of PKC epsilon induced by TNF alpha enables this PKC isoform to interact with insulin signaling and to inhibit the insulin receptor kinase activity. PMID- 9414110 TI - Determination of fast proton exchange rates of biomolecules by NMR using water selective diffusion experiments. AB - We present a new water selective pulse sequence allowing rapid determination of exchange rates of labile protons on the millisecond time scale. Using diffusion measurements, exchange rates of resolved protons can be determined without prior knowledge of relaxation parameters in a short overnight experiment. The use of a sensitive, highly selective and easy to implement water excitation scheme allows for its straightforward application to a wide range of biomolecules. The results obtained for the imino proton exchange rates of a 16 bp DNA are in strong agreement with values obtained by the classical approach of two-dimensional exchange spectroscopy. PMID- 9414109 TI - The C-terminus of yeast plasma membrane H+-ATPase is essential for the regulation of this enzyme by heat shock protein Hsp30, but not for stress activation. AB - Several stresses cause additional activation to the glucose-stimulated plasma membrane H+-ATPase activity of yeast, but it is not clear how this is achieved. We recently reported that cells lacking the integral plasma membrane heat shock protein Hsp30 display enhanced increases in plasma membrane H+-ATPase activity with either heat shock or weak organic acid stress (Piper, P.W., Ortiz-Calderon, C., Holyoak, C., Coote, P. and Cole, M. (1997) Cell Stress and Chaperones 2, 12 24), indicating that the stress induction of Hsp30 acts to reduce stress stimulation of the H+-ATPase. In this study it is shown that Hsp30 acts to reduce the Vmax of H+-ATPase in heat shocked cells. Its action is lost with deletion of the C-terminal 11 amino acids of the H+-ATPase, a deletion that does not abolish the stress stimulation of this enzyme. Mutation of the Thr-912 residue within the C-terminal domain of H+-ATPase, a potential site of phosphorylation by the Ca2+ calmodulin-dependent protein kinase, also abolishes any effect of Hsp30. Hsp30 may therefore be acting on a Thr-912 phosphorylated form of the H+-ATPase. PMID- 9414111 TI - Cyclosporin A-sensitive release of Ca2+ from mitochondria in intact thymocytes. AB - Release of Ca2+ from mitochondria into cytosol in intact thymocytes was studied using the fluorescent dye Fluo-3. It was shown that the release of Ca2+ induced by the dithiol cross-linking agent phenylarsine oxide or by uncoupler was strongly inhibited by cyclosporin A, a specific inhibitor of the permeability transition pore (PTP) in mitochondria. Oxidative stress sensitized the pore so even partial uncoupling caused rapid cyclosporin A-sensitive release of Ca2+. The experiments on digitonin-permeabilized cells confirmed that uncoupling induced opening of the PTP, which forms the major pathway for rapid release of Ca2+ from thymocyte mitochondria. PMID- 9414112 TI - Superinduction of COX-2 mRNA by cycloheximide and interleukin-1beta involves increased transcription and correlates with increased NF-kappaB and JNK activation. AB - Many primary response genes, including cyclooxygenase-2 (COX-2), exhibit mRNA superinduction following agonist stimulation in the presence of translational blockers such as cycloheximide. This is widely assumed to result from mRNA stabilisation. However, superinduction of IL-1beta-induced COX-2 mRNA levels by cycloheximide in pulmonary type II A549 cells occurred by increased transcription and not by mRNA stabilisation. Furthermore, equivalent effects were observed on NF-kappaB binding to COX-2 promoter kappaB sites and activation of the Jun N terminal kinases (JNK), p54 and p46. These signalling pathways play important roles in COX-2 induction and may therefore account for the observed increases in COX-2 transcription. These data are consistent with negative feed-back involving down-regulation of NF-kappaB by de novo IkappaB alpha synthesis and suggest that JNK activation may also be down-regulated by a cycloheximide sensitive process. PMID- 9414113 TI - In vitro evidence that hsp90 contains two independent chaperone sites. AB - Hsp90 is an abundant and constitutively expressed stress protein and molecular chaperone. Here we dissected human hsp90 into three major domains to identify the putative chaperone site at which hsp90 binds unfolded polypeptide. Surprisingly, both the N-terminal and the C-terminal domain of hsp90 prevent the aggregation of denatured polypeptides. The chaperone activity of the N-domain is inhibited by geldanamycin, a specific inhibitor of hsp90-mediated protein refolding. While both domains suppress protein aggregation, only the C-domain binds an antigenic peptide derived from VSV G. Based on these results, hsp90 may be the first chaperone to contain two independent chaperone sites with differential specificity. PMID- 9414114 TI - Selective activation of JNK/SAPK by interleukin-1 in rabbit liver is mediated by MKK7. AB - Activation of jun N-terminal kinase (JNK)/stress-activated protein kinase (SAPK) by interleukin-1 (IL-1) has been reported in many cells and in rabbit liver. Here we report selective activation of JNK/SAPK, without activation of p38 or p42 mitogen-activated protein kinases (MAPKs), by IL-1 in rabbit liver. We identified an IL-1 regulated JNK/SAPK activator present in rabbit liver using S Sepharose chromatography. It was purified and immunoprecipitated by two antisera to MAP kinase kinase 7 (MKK7). It was not recognised by an antibody to MKK4. We conclude that MKK7 is the activator of JNK/SAPK activated by IL-1 in liver and that JNK/SAPK is the only MAPK activated by IL-1 in liver. PMID- 9414115 TI - Activation of mitogen-activated protein kinase is involved in sphingosine 1 phosphate-stimulated interleukin-6 synthesis in osteoblasts. AB - We previously showed that sphingosine 1-phosphate (SPP) acts as a second messenger for tumor necrosis factor alpha-induced interleukin-6 (IL-6) synthesis in osteoblast-like MC3T3-E1 cells. In the present study, we further investigated the mechanism of IL-6 synthesis induced by SPP in MC3T3-E1 cells. SPP significantly induced p42/p44 mitogen-activated protein (MAP) kinase activity. PD98059, an inhibitor of MAP kinase kinase, suppressed SPP-induced IL-6 synthesis as well as SPP-induced MAP kinase activation. The patterns of both inhibitions were similar. TMB-8, an inhibitor of Ca2+ mobilization from intracellular Ca2+ stores, significantly suppressed the SPP-induced IL-6 synthesis. These results strongly suggest that SPP-induced IL-6 synthesis is mediated via p42/p44 MAP kinase activation in osteoblast-like cells and that the SPP-induced IL-6 synthesis is dependent on intracellular Ca2+ mobilization. PMID- 9414116 TI - Relative bioavailability of the antioxidant flavonoid quercetin from various foods in man. AB - Quercetin is a strong antioxidant and a major dietary flavonoid. Epidemiological studies suggest that consumption of quercetin protects against cardiovascular disease, but its absorption in man is controversial. We fed nine subjects a single large dose of onions, which contain glucose conjugates of quercetin, apples, which contain both glucose and non-glucose quercetin glycosides, or pure quercetin-3-rutinoside, the major quercetin glycoside in tea. Plasma levels were then measured over 36 h. Bioavailability of quercetin from apples and of pure quercetin rutinoside was both 30% relative to onions. Peak levels were achieved less than 0.7 h after ingestion of onions, 2.5 h after apples and 9 h after the rutinoside. Half-lives of elimination were 28 h for onions and 23 h for apples. We conclude that conjugation with glucose enhances absorption from the small gut. Because of the long half-lives of elimination, repeated consumption of quercetin containing foods will cause accumulation of quercetin in blood. PMID- 9414117 TI - Identification of the transcription start site for the spinach chloroplast serine tRNA gene. AB - Deleting part of the 3' end of the spinach chloroplast serine tRNA coding region, which destroyed the proper folding of its RNA transcript and resulted in the inhibition of tRNA processing, allowed the detection of a serine tRNA primary transcript. The transcription start site for this primary transcript, synthesized from the internal promoter, was mapped to -12 upstream from the mature tRNA coding region. Transcription analysis with various 5' deletion mutants suggested that the AT-rich region between -31 and -11, immediately upstream of the serine tRNA transcription start site, affects the transcription efficiency, and possibly the selection of transcription start site. Identification of the transcription start site for the spinach chloroplast serine tRNA gene in this study represents the first example of 5' end mapping of a tRNA precursor transcribed from chloroplast tRNA genes containing an internal promoter. PMID- 9414119 TI - Mitochondrial DNA deletions in oculopharyngeal muscular dystrophy. AB - The deletions in the mitochondrial DNA from skeletal muscle samples of two oculopharyngeal muscular dystrophy cases were studied using polymerase chain reaction techniques. The 4977 bp 'common deletion' was present in both specimens, exceeding the corresponding values of similarly aged, healthy controls. In the two samples multiple different mitochondrial DNA deletions, some case-specific and present at quite high, although not pathogenetic levels, were observed. The results suggest that mitochondrial DNA deletions, and the 'common deletion' in particular, might be a sensitive and early marker of a generalized mitochondrial suffering, due to a variety of pathological and physiological causes. PMID- 9414118 TI - Proteolytic processing of presenilin-1 (PS-1) is not associated with Alzheimer's disease with or without PS-1 mutations. AB - Cerebral presenilin-1 protein (PS-1) is normally composed of the amino-terminal fragment (NTF) with Mr 28 kDa and the carboxy-terminal fragment (CTF) with 18 kDa. We analyzed human PS-1 in brains with early-onset familial Alzheimer's disease (FAD) with and without PS-1 mutations to study whether mutated PS-1 was abnormally metabolized. Cerebral PS-1 were found to be cleaved into two fragments of NTF and CTF independently of the occurrence of PS-1 mutation in human brains. A small portion of PS-1 was recently found to suffer another processing by caspase-3, an apoptosis-related cysteine protease. In contrast to the recent finding that the Volga-German mutation on presenilin-2 (PS-2) affects the increasing caspase-3 PS-2 fragment, the PS-1 mutation did not cause a significant change in PS-1 fragmentation. We conclude that PS-1 fragmentation and other (probably caspase-3-mediated) digestion following apoptosis occur independently of PS-1 mutations. PMID- 9414120 TI - Induction of UCP2 mRNA by thyroid hormones in rat heart. AB - The possible regulation of the expression of uncoupling protein-2 (UCP2) mRNA by thyroid hormones in different tissues was examined in rats. Triiodothyronine (T3) was found to produce an organ-specific enhancement of UCP2 expression in rat tissues. The effect of T3 was markedly observed in heart, whereas a moderate effect was seen in skeletal muscle and no effect in kidney or liver. These results suggest that UCP2 is a protein that may be involved in the nuclear mediated effect of T3 on resting metabolic rate in the rat. PMID- 9414121 TI - Involvement of NF-kappaB in the regulation of cyclooxygenase-2 protein expression in LPS-stimulated J774 macrophages. AB - We investigated the involvement of NF-kappaB in the regulation of COX-2 protein expression and prostaglandin production in LPS-stimulated J774 macrophages. Incubation of J774 cells with LPS (1 microg/ml) for 24 h caused an increase of COX-2 protein expression and accumulation of both PGE2 and 6-keto-PGF1alpha in the cell culture medium. Ammonium pyrrolidinedithiocarbamate (APDC, 0.1, 1, 10 microM) and N-alpha-p-tosyl-L-lysine chloromethylketone (TLCK, 1, 10, 100 microM), two inhibitors of NF-kappaB activation, suppressed in a concentration dependent manner both LPS-induced COX-2 protein expression and prostanoid generation. Moreover, APDC and TLCK both inhibited the LPS-induced increase of NF kappaB DNA binding activity and prevented IkappaB-alpha degradation. Our results show for the first time that NF-kappaB is involved in COX-2 protein expression in LPS-stimulated J774 macrophages and suggest that inhibitors of NF-kappaB activation may represent a useful tool for the pharmacological control of inflammation. PMID- 9414122 TI - Ceramide-activated protein phosphatase-2A activity in insulin-secreting cells. AB - Okadaic acid (OKA)-sensitive phosphatase (PP2A) activity may modulate nutrient induced insulin secretion from pancreatic beta cells [Kowluru et al., Endocrinology 137 (1996) 2315-2323]. Ceramides, a new class of lipid second messengers may regulate PP2A [Dobrowsky and Hannun, J. Biol. Chem. (1992) 267, 5048-5051], and might play a role in cytokine-mediated apoptosis in beta cells [Sjoholm, FEBS Lett. 367 (1995) 283-286]. Therefore, we investigated the regulation of PP2A-like activity by ceramides in isolated beta (HIT-T15 or INS-1) cells. Cell-permeable (C2, C6 or C18) ceramides stimulated OKA-sensitive (but not -insensitive) phosphatase activity in a concentration-dependent manner (0-12.5 microM), with maximal stimulation (+50-100%) at < 12.5 microM. C2-dihydroceramide (a biologically inactive analog of C2 ceramide) failed to augment PP2A-like activity. Stimulatory effects of ceramides do not appear to be mediated via activation of the carboxyl methylation of the catalytic subunit of protein phosphatase 2A, since no effects of ceramides (up to 25 microM) were demonstrable on this parameter. These data identify a ceramide-activated protein phosphatase as a possible locus at which ceramides might exert their effects on beta cells leading to altered insulin secretion, and decreased cell viability followed by apoptotic cell demise. PMID- 9414123 TI - Genomic structure of a potassium channel toxin from Heteractis magnifica. AB - We provide information on the gene encoding the K+ channel toxin, HmK, of the sea anemone Heteractis magnifica. A series of DNA amplifications by PCR, which included the amplification of the 5'-untranslated region of the gene, showed that an intron of 402 nucleotides separated the sequence that encodes the matured toxin from the signal peptide sequence. A second 264 nucleotide intron interrupted the 5'-untranslated region of the previously reported HmK cDNA. Two possible transcription-initiation sites were identified by primer extension analysis. Corresponding TATA-box consensus sequences, characteristic of a promoter region, were also located from PCR products of uncloned libraries of adaptor-ligated genomic DNA fragments. The coding region for matured HmK is intronless. The same is also true for other sea anemone toxins reported thus far. More notably, a similar intron-exon organization is present in other ion channel blocking toxins from scorpions implying that molecules having similar functions share a similar organization at the genomic level suggesting a common path of evolution. PMID- 9414124 TI - Dissecting the assembly pathway of the 20S proteasome. AB - Proteasomes reach their mature active state via a complex cascade of folding, assembly and processing events. The Rhodococcus proteasome offers a means to dissect the assembly pathway and to characterize intermediates; its four subunits (alpha1, alpha2, beta1, beta2) assemble efficiently in vitro with any combination of alpha and beta. Assembly studies with wild-type and N-terminally truncated beta-subunits in conjunction with refolding studies allowed to define the role of the propeptide which is two-fold: It supports the initial folding of the beta subunits and it promotes the maturation of the holoproteasomes. PMID- 9414125 TI - Primary structure and expression of a naturally truncated human P2X ATP receptor subunit from brain and immune system. AB - A novel member of the ionotropic ATP receptor gene family has been identified in human brain. This 422 amino acid long P2X receptor subunit has 62% sequence identity with rat P2X5. Several characteristic motifs of ATP-gated channels are present in its primary structure, but this P2X5-related subunit displays a single transmembrane domain. Heterologous expression of chimeric subunits containing the C-terminal domain of rat P2X5 leads to the formation of desensitizing functional ATP-gated channels in Xenopus oocytes. The developmentally regulated mRNA, found in two splicing variant forms, is expressed at high levels in brain and immune system. PMID- 9414126 TI - Cloning of rat uncoupling protein-3 and uncoupling protein-2 cDNAs: their gene expression in rats fed high-fat diet. AB - In order to elucidate energy balance in the skeletal muscle, we cloned cDNA of a homologue of uncoupling protein (UCP) from rat skeletal muscle. We also cloned rat UCP-2 cDNA from rat brown adipose tissue (BAT). The UCP cloned from rat skeletal muscle showed 57% and 72% identity with rat UCP-1 and UCP-2. The mRNA was expressed abundantly in the skeletal muscle, moderately in the BAT, and slightly in the white adipose tissue (WAT) with a major band at 2.5 kb and a minor band at 2.8 kb, while the UCP-2 gene expression was widely detected in the whole body with substantial levels in the WAT and with slight levels in the skeletal muscle and BAT. The rat UCP cloned in the present study showed 86% identity with the recently cloned human UCP-3, which was also expressed abundantly in the skeletal muscle with a signal of 2.4 kb. Therefore, the rat UCP was considered to be rat UCP-3. In rats fed high-fat diet the UCP-3 gene expression was augmented 2-fold in the skeletal muscle while UCP-2 mRNA levels were increased significantly (1.6-fold) in the epididymal WAT. Augmented expression of UCPs may provide defense against high-fat induced obesity and impairment of glucose metabolism. PMID- 9414127 TI - Detection and cloning of unique integration sites of retrotransposon, intracisternal A-particle element in the genome of acute myeloid leukemia cells in mice. AB - We previously found retrotransposition of the intracisternal A-particle (IAP) element in the genome of acute myeloid leukemia (AML) cells induced by X irradiation of C3H/He mice (FEBS 16333). To analyze the occurrence of the IAP mediated retrotransposition in AML cells, we compared integration sites of the IAP element by polymerase chain reaction (PCR) in the genomes of five AML strains derived from different C3H mice. Unique PCR products were found in all of the above independent leukemia cells, whereas no such products were detected in normal cells. Results of cloning, sequencing and Southern analyses showed that the PCR products were derived from novel integration sites of the IAP element in the genome. The data suggest that IAP-mediated retrotransposition occurs frequently in radiation-induced AML cells from C3H/He mice. PMID- 9414128 TI - PPP1R6, a novel member of the family of glycogen-targetting subunits of protein phosphatase 1. AB - A complementary DNA encoding a novel human protein phosphatase 1 (PP1) glycogen targetting subunit of molecular mass 33 kDa has been sequenced. PPP1R6 is 31% identical to the glycogen-targetting subunit (G(L)) of PP1 from rat liver, 28% identical to the N-terminal region of the glycogen-targetting subunit (G(M)) from human skeletal muscle and 27% identical to glycogen-targetting subunit PPP1R5. Unlike human PPP1R5 and its murine homologue PTG, whose mRNAs are most abundant in skeletal muscle, heart and liver, PPP1R6 is present at similar levels in a wide variety of tissues. The PPP1R6 is associated with glycogen in muscle but is not subject to the same modes of covalent and allosteric regulation as G(M) and G(L). PMID- 9414129 TI - c-Rel and p65 subunits bind to an upstream NF-kappaB site in human granulocyte macrophage-colony stimulating factor promoter involved in phorbol ester response in 5637 cells. AB - To further clarify the complex transcriptional regulation of the human GM-CSF gene, which was extensively investigated in activated T cells, we have studied the role of an upstream NF-kappaB like site in the 5637 non-lymphoid cell line, which derives from a bladder carcinoma and constitutively produces GM-CSF. This sequence, named the A element, has an active role on GM-CSF transcription and is responsive to the tumor promoter PMA in transient transfection experiments. We describe here a heterodimeric binding complex of NF-kappaB subunits (c-Rel and p65) which is identical to the one obtained using the HIV-LTR-kappaB site as recognition sequence and different from the one (c-Rel and p50) observed with nuclear extracts from Mo T-lymphoid HTLV-II infected cells. PMID- 9414130 TI - Preparation of peptides which mimic glycosphingolipids by using phage peptide library and their modulation on beta-galactosidase activity. AB - We describe the use of a phage-displayed random pentadecamer peptide library for searching glycosphingolipid mimicking peptides. Two phage clones (AD-1 and AD-2) were selected by biopanning using monoclonal antibody AD117m, directed to lactotetraosylceramide (Lc4Cer). The amino acid sequences of the selected clones showed high homology (VPPXFXXXY) in 9-mer. Three phage clones were selected by using monoclonal antibody H11, directed to neolactotetraosylceramide (nLc4Cer), the linkage isomer of Lc4Cer, and the displayed amino acid sequences were compared. One of these peptides showed the same amino acid sequence as that of AD 2 except for one amino acid substitution. Pentadecamer, 9-mer and point mutated 9 mer peptides were synthesized on the basis of the displayed amino acid sequences. Binding activity of the peptides to the monoclonal antibodies or Ricinus communis lectin showed that 9-mer peptides are enough to mimic the epitope carbohydrate structure. Furthermore, six of the synthesized peptides inhibited Jack bean beta galactosidase activity towards nLc4Cer at a high concentration of the enzyme, whereas at lower enzyme concentrations some peptides showed potent activation of the enzyme activity. This is the first report of carbohydrate mimicking peptides which modulate glycosidase activity. PMID- 9414131 TI - Cyclolinopeptide A (CLA) mediates its immunosuppressive activity through cyclophilin-dependent calcineurin inactivation. AB - The immunosuppressive cyclic nonapeptide cyclolinopeptide A inhibits calcium dependent, but not calcium-independent, activation of T lymphocytes comparably to the actions of cyclosporin A and FK506. The concentration required for complete inhibition, however, is 10 times higher than that of cyclosporin A. In addition, we demonstrate that calcineurin, a phosphatase which plays an important role in T lymphocyte signalling, is inhibited in vitro by cyclolinopeptide A by a mechanism dependent on the peptidyl-prolyl cis-trans isomerase (PPIase) cyclophilin A but not FKBP12. Direct binding of cyclolinopeptide A to cyclophilin A was confirmed using tryptophan fluorescence studies and PPIase assays. These results represent a third example of the production of a natural product that neutralises calcineurin by a mechanism dependent on the primary binding to a PPIase. PMID- 9414132 TI - Immunologic consequences of blood transfusion and their clinical manifestations. AB - Aspects of clinical immunology, in the context of transfusion medicine, became highlighted in the last decade as a consequence of the accelerating expansion of basic immunology, immunogenetics and molecular biology. In addition sophisticated new technologies, which were capable of producing pure and safe blood products, attracted more attention to research and monitor the consequences of transfusion. These technologies also had obvious effects on supportive hematological therapy. The transfusion of blood components follows the rules of organ transplantation: when there is a mismatch between the donor and the recipient, the transfusion has the potential to induce various kinds of immune response against alloantigens. Antigen-compatible transfusions that involve major and rare blood groups are in almost all cases mismatched with respect to various polymorphic systems expressed on the cellular blood components. These include histocompatibility leukocyte antigens (HLA), tissue-specific and differentiation alloantigens, and, in the case of plasma, immunoglobulins, complement components, heat shock proteins, and shedded soluble membrane alloantigens. Clinical manifestations of alloimmune responses are typically deleterious. For example, immediate antigen-antibody binding and its consequences as secondary activations are paralleled by the nonhaemolytic febrile reaction, HLA sensitization can lead to a state of platelet refractoriness and inconvenient clinical symptoms. In certain immunogenetic situations and in immunodeficient patients graft-versus-host disease can be induced by blood products that contain live lymphocytes. Leukocyte filtration techniques are widely used to avoid most but not all of these harmful side effects of blood component therapy. In contrast to these harmful side effects in certain immunogenetic conditions, alloantigens that are expressed on various blood products can elicit an advantageous suppression of the immune response in the recipient. In the context of kidney transplantation this is termed the 'beneficial transfusion effect', and typically results in the prolongation of the graft's survival. In cases of recurrent habitual abortion and IgG therapy associated with certain autoimmune diseases, immunization with leukocytes specifically takes advantage of this phenomenon. To date the beneficial transfusion effect is not fully understood. In certain cases of malignancies or gastrointestinal surgeries this suppression of immune regulation that is induced by transfusion can worsen the clinical state either by permitting the spread of the tumor or by allowing severe infections to proceed unchecked. In conclusion it is imperative to monitor the immunological consequences of transfusion in order to deter the disadvantageous side effects. Taking advantage of the 'beneficial transfusion effect' may also provide a new means for immune therapy using the various blood products. PMID- 9414134 TI - In vitro inhibitory activity of tumor necrosis factor alpha and interleukin-2 of human immunoglobulin preparations. AB - Human immunoglobulin preparations have been used in a number of clinical settings with good results, although in many of them the mechanism of action is not yet known. One possible mechanism is the modulation of cytokine activity. This study investigated the presence of inhibitory activity in intravenous immunoglobulin (IVIg) and F(ab')2 fragment preparations to two cytokines, tumor necrosis factor alpha (TNF-alpha) and interleukin-2 (IL-2). Cytotoxic activity of human recombinant TNF-alpha or TNF-alpha secreted by peripheral blood mononuclear cells (PBMC) on L929 cells and the proliferative activity of the IL-2 on CTLL-2 cells were examined. Human serum albumin (HSA) was used as control. F(ab')2 inhibited, in a dose-dependent fashion, the TNF-alpha activity secreted by PBMC serial dilutions or, at the higher concentrations (25 and 10 mg/ml), recombinant TNF alpha activity. In contrast, IVIg was able to inhibit only at 25 and 10 mg/ml the TNF-alpha activity secreted by any PBMC dilution tested, and did not inhibit the recombinant TNF-alpha activity. With IL-2, however, even HSA was able to inhibit its proliferative activity, possibly through a carrier effect. The IVIg inhibition of IL-2 activity was not different from that of HSA, but F(ab')2, at 12.5 mg/ml, was capable of inhibiting significantly more the IL-2 activity than HSA. Our results suggest an anticytokine effect of the immunoglobulin preparations that this activity may be mainly mediated by variable regions of the immunoglobulins, and that the more pronounced effect of F(ab')2 may be due to its greater molar concentration compared to intact IgG molecules. PMID- 9414133 TI - A system for simultaneous evaluation of the effect of signal transduction inhibitors on interleukin-2 synthesis and cell viability in a human T cell line. AB - BACKGROUND: The signal transduction pathways involved in interleukin-2 (IL-2) synthesis have become a focus of interest for pharmacological intervention. Regulation of synthesis of IL-2 requires costimulation of two cell surface receptors, the T cell receptor and an appropriate costimulatory receptor. Antibodies such as anti-CD3 to stimulate the T cell receptor and anti-CD28 can be used to induce this costimulation. Previous methodologies employed antibody coupled to polystyrene microtiter plates or solution phase stimulation. METHODS: Here, a method using magnetic beads to present anti-CD3 and anti-CD28, to the Jurkat T cell line was developed. Conditions were also developed for simultaneous analysis of the effect of test compounds on cell viability. The IL-2 synthesis methods and cell viability methods have been used to evaluate signal transduction inhibitors. RESULTS: This method was then employed to test the effects of various signal transduction inhibitors on IL-2 synthesis. Agents which increase cAMP, cyclosporin and inhibitors of calmodulin, tyrosine kinases/phosphatases, protein kinase C, and PC-PL-C decrease IL-2 synthesis at concentrations which do not affect cell viability. CONCLUSION: These results found with this novel system of stimulation with antibody to CD3 and CD28 compare favorably with activities reported in the literature thus validating this rapid facile system for evaluation of the effect of signal transduction inhibitors in IL-2 synthesis. PMID- 9414135 TI - Effects of tumor necrosis factor and pentoxifylline on ICAM-1 expression on human polymorphonuclear granulocytes. AB - Intercellular adhesion molecule-1 (ICAM-1) is a cytokine-inducible adhesion molecule, expressed on cells of multiple lineages at the site of inflammation. Cytofluorometric analysis revealed that CD16-positive peripheral blood polymorphonuclear leukocytes (PMNs, neutrophils) expressed ICAM-1 on their surface, and it was upregulated by in vitro stimulation with tumor necrosis factor (TNF), GM-CSF and Staphylococcus aureus. The S. aureus-induced stimulation of ICAM-1 expression was inhibited by pentoxifylline (PTX). As TNF is a potent inducer of ICAM-1 expression, it is concluded that in these experiments the inhibition of TNF production by PTX concomitantly resulted in the inhibition of the upregulation of ICAM-1. However, the inhibition of granulocyte apoptosis by PTX might be of importance in this process. The present study provides evidence that cytokine-stimulated neutrophils are able to express the adhesion molecule ICAM-1 and this may allow ICAM-1-positive neutrophils to physically interact with LFA-1-positive inflammatory cells. The preliminary results demonstrate that the basal expression of ICAM-1 on PMNs of septic patients is higher than that in the case of normal blood donors. Further studies will elucidate the in vivo relevance of cytokine-induced neutrophil ICAM-1 expression and the potential role of its inhibition by PTX in inflammatory response disorders. PMID- 9414136 TI - The role of nitric oxide in human pulmonary artery endothelial cell injury mediated by neutrophils. AB - Human endothelial cells are injured by the action of leukocytes. We investigated the role of nitric oxide (NO) in the induction of injury to human pulmonary artery endothelial cells. NO has been a putative source of cytotoxic reactive oxygen species in some settings. Incubation of endothelial cells with neutrophils increased the release of lactate dehydrogenase activity and preloaded fura-2 from endothelial cells, indicating that neutrophils induce endothelial cell injury. This effect was augmented by treatment with carboxy-PTIO, which traps NO in the medium, or with L-NAME, an inhibitor of NO synthase. When endothelial cells were incubated with neutrophils stimulated by phorbol myristate acetate, an activator of protein kinase C, endothelial cell damage was further enhanced and the amount of NO in the medium was decreased. Dibutyryl cyclic AMP, a cell-permeable analogue of cyclic AMP, protected against neutrophil-induced endothelial cell injury and increased NO release into the medium. The effects of dibutyryl cyclic AMP were abrogated by treatment with H-89, a potent inhibitor of cyclic AMP dependent protein kinase. The protective effect on neutrophil-induced endothelial cell injury by dibutyryl cyclic AMP was abolished by addition of carboxy-PTIO or L-NAME. Thus, our studies suggest that NO, presumably released from endothelial cells, protects against endothelial injury by activated neutrophils and the protective effect by cyclic AMP during coculture with activated neutrophils is mediated through the action of NO. However, when monocytes activated by lipopolysaccharide and IFN-gamma were used instead of neutrophils, endothelial cells were likewise injured, but a much higher level of NO was detected and injury was diminished by addition of carboxy-PTIO to the medium. These observations suggest that the high levels of NO released by activated monocytes contribute to endothelial injury, whereas low levels of NO protect endothelial cells against injury by neutrophils. PMID- 9414137 TI - Tissue suspension agglutination: a simple method to screen species-specific and organ-specific reactions. AB - Agglutination tests with preparations of parenchymatous organs were developed. The tissue suspensions were dried at room temperature after they had been spread as a very thin layer on a glass plate, or otherwise, they were lyophilized. The dried preparations were pulverized and then prepared as stable suspensions in saline. The agglutination test was conducted on a slide by mixing one drop of the tested serum at a convenient dilution with one drop of tissue powder suspension. Agglutination in the form of readily discernible clumps could be assessed after 1 10 min. By means of this procedure, species-specific reactions were studied using suspensions of kidneys of various species. Organ-specific reactions were noted with suspensions of brain and thyroid. Agglutination of thyroid powder was observed with rabbit anti-rabbit thyroid sera as well as with many, albeit not all, sera of patients with Hashimoto's disease. PMID- 9414138 TI - Cyclic AMP-elevating agents inhibit mite-antigen-induced IL-4 and IL-13 release from basophil-enriched leukocyte preparation. AB - Human peripheral blood basophils are known to secrete interleukin (IL)-4 and IL 13 after cross-linking of cell surface IgE. However, little is known about the pharmacological regulation of allergic cytokine release from basophils. In the present study, we investigated the effects of cyclic 3',5'-adenosine monophosphate (cAMP)-elevating agents on antigen-induced IL-4 and IL-13 release from basophil-enriched leukocyte preparations. We obtained venous blood from 27 atopic asthmatic patients (mean age was 45.8+/-3.6 years, all patients were sensitive to mite antigen) and prepared basophil-enriched leukocyte preparations by double-Percoll gradients (basophil purity was 13.4+/-1.6%). The cell preparations were treated with phosphodiesterase (PDE) inhibitors, dexamethasone, forskolin or dibutyryl cAMP for 10 min and were challenged with mite antigen for 6 h. The released IL-4 and IL-13 in the supernatants were measured by enzyme linked immunosorbent assay systems. No IL-4 or IL-13 was detected in the supernatant of the basophil-depleted preparation after the challenge with mite antigen, suggesting that basophils specifically produce these cytokines. A nonselective PDE inhibitor, theophylline, and a PDE IV-selective inhibitor, rolipram, significantly suppressed the release of IL-4 and IL-13 from the basophil-enriched preparation. Although several concentrations of cilostazol, a PDE III-selective inhibitor, had no effect on the release of both cytokines, cilostazol suppressed the release of IL-4 additively when applied with rolipram. Forskolin and dibutyril cAMP also significantly suppressed the release of both cytokines, suggesting that the suppressive effects by PDE inhibitors were accompanied by the elevations in cAMP levels. We conclude that basophil-enriched leukocyte preparations produce IL-4 and IL-13 in response to antigen and that the release of these cytokines could be regulated by cAMP-modulating agents. PMID- 9414139 TI - Identification of Dermatophagoides farinae-2-derived peptides and class II HLA molecules recognized by T cells from atopic individuals. AB - Der f2, the group 2 allergen of Dermatophagoides farinae, is one of the major inhalation allergens in Japan. Using the mixture of a panel of overlapping synthetic peptides that spread over the entire Der f2 molecule, we found that polyclonal Der f2-specific short-term T cell lines prepared from peripheral blood mononuclear cells of 5 individuals allergic to Der f who carry most of the common HLA haplotypes seen in the Japanese population can respond mainly to 7 different peptides. Distribution of the T cell epitopes on Der f2 was not identical with that on Der p2. Five of 7 peptides stimulated T cells of more than 2 donors, regardless of HLA types. Inhibition patterns by anti-HLA class II mAbs were heterogenous in proliferative responses of each cell line, three were mainly inhibited by anti-HLA-DR mAb, and the others were inhibited by anti-HLA-DQ mAb. One of these T cell lines, SM, of which the proliferative response was partially inhibited by anti-HLA-DQ mAb, was cloned. Indeed, the T cell clone SM4.6 was restricted by DQ6 molecules encoded for by an HLA-DRB1*1502-DRB5*0102-DQA1*0103 DQB1*0601 haplotype. These results indicate that patients' T cells recognize Der f2 in association with a variety of HLA-DR or HLA-DQ as antigen-presenting molecules. Thus, although some peptides do have a more potent T cell stimulatory activity than others, the TCR ligands formed with the Der f2 molecule are highly heterogeneous, a factor also noted in Der f1-specific T cell lines in our previous study. PMID- 9414140 TI - Sulfidoleukotriene release test (CAST) in hypersensitivity to nonsteroidal anti inflammatory drugs. AB - There is a great need to develop a method for making an accurate and reliable in vitro diagnosis of adverse hypersensitivity reactions to drugs. We measured the amount of sulfidoleukotriene (sLT) released from the peripheral blood leukocytes obtained from 25 patients who developed hypersensitivity reactions following the administration of nonsteroidal anti-inflammatory drugs (NSAIDs); 12 patients demonstrated reactions to Voltaren, 8 patients to Bufferin, and 5 patients to Sedes G. The stimulation index, the ratio of the amounts of sLT (pg/ml) incubated with and without drugs, was considerably higher in the patients than in the controls, which consisted of 5 nonallergic healthy subjects. The sensitivity of the CAST (cellular antigen stimulation test) was evaluated to range from 62.5 to 80%, while the specificity was 70-100%. The CAST may thus be useful as a novel in vitro test system in order to screen for possible hypersensitive reactions to NSAIDs with both reliability and safety. PMID- 9414141 TI - Seasonal increase in human IgE and IgG4 antisaliva antibodies to Aedes mosquito bites. AB - BACKGROUND: Mosquito bite-sensitive subjects frequently have circulating IgE and IgG4 antibodies to Aedes mosquito saliva proteins. METHODS: In the present study we examined the antibody response during a mosquito season in 14 subjects living in Finnish Lapland. Immunoblotting was performed with Aedes communis saliva and the 22- and 36-kD antisaliva antibody bands were analyzed. RESULTS: The preseason sera showed IgE antibodies to the main saliva antigens in 12, IgG4 antibodies in all 14 and IgG1 antibodies in 12 subjects, and the postseason sera in all but 1 subject. The postseason sera showed significantly more intense IgE (p < 0.05), IgG4 (p < 0.001) and IgG1 (p < 0.01) antibody bands than the preseason sera. CONCLUSION: These results show that seasonal exposure to mosquito bites leads to an increased IgE, IgG4 and IgG1 antibody response, a phenomenon similar to that occurring e.g. in pollen allergy. PMID- 9414144 TI - Immunosuppressive effect of honey on the induction of allergen-specific humoral antibody response in mice. AB - Our study with honey for its possible immunomodulatory activity reveals the immunosuppressive activity on induction of murine humoral antibody responses against different allergens as determined by passive cutaneous anaphylaxis and Ouchterlony double immunodiffusion techniques. Ovalbumin (OVA)-specific IgE antibody responses elicited with various doses were completely suppressed by different sources of commercial honeys. Honey is also found to have suppressed the induction of OVA-specific humoral antibody responses in different strains of mice. The results obtained in this work confirm the immunosuppressive activity of honey and suggest its possible applicability in conditions requiring immunosuppression. PMID- 9414142 TI - Urinary excretion of leukotriene E4 and 11-dehydrothromboxane B2 in patients with spontaneous asthma attacks. AB - BACKGROUND: Cysteinyl leukotrienes (LTs) and thromboxane A2 (TXA2) are known to play an essential role in the pathogenesis of atopic asthma. However, their role in nonatopic asthma has not as yet been clarified. The objectives of this study were to define (1) the participation of LTs and TXA2 in nonatopic asthma and (2) the relationship between LTs and TXA2 in asthma attacks. METHODS: Urinary excretion of leukotriene E4 (LTE4) and 11-dehydrothromboxane B2 (11DTXB2) was measured in 10 atopic and 10 nonatopic asthmatics who were admitted to hospital with either an acute asthma attack or status asthmaticus. RESULTS: In atopic asthmatics, urinary excretion of LTE4 and 11DTXB2 was significantly higher on admission with an asthma attack, and returned to control levels when the patients were in the improved state (179+/-29 to 65+/-16 ng/day in LTE4, 1,085+/-250 to 440+/-90 ng/day in 11DTXB2). Similar findings were observed in nonatopic asthmatics (148+/-13 to 61+/-11 ng/day in LTE4, 1,089+/-206 to 457+/-60 ng/day in 11DTXB2). However, when the individual data during the attack were analyzed, there was no correlation between urinary excretion of LTE4 and that of 11DTXB2 in both types of asthma. CONCLUSION: Both LTs and TXA2 may be implicated in the pathogenesis of the nonatopic as well as the atopic type of asthma, but no correlation between these two metabolites was observed in the individuals. PMID- 9414143 TI - Effect of a phosphodiesterase 3 inhibitor, cilostazol, on bronchial hyperresponsiveness in elderly patients with asthma. AB - BACKGROUND: Over the last few years, evidence has accumulated that cyclic nucleotide phosphodiesterase (PDE) 3 and/or PDE4 inhibitors may be useful for the treatment of asthma. The present study was designed to examine the effect of a selective orally active PDE3 inhibitor, cilostazol, on bronchial hyperresponsiveness (BHR) of asthma. METHODS: The effect of a single oral dose of cilostazol on BHR was studied in 10 elderly patients with clinically stable asthma, with a mean age of 59.5+/-4.8 years, in a double-blind, crossover, placebo-controlled study. Each subject received 100 mg of cilostazol, 200 mg of theophylline as a positive control or a placebo in a random order. The subjects underwent a methacholine challenge test 3 h after each drug administration on three occasions separated by 2 weeks. RESULTS: The geometric mean value of the provocative concentration of methacholine causing a 20% fall in forced expiratory volume in 1 s (PC20-FEV1) with cilostazol was 0.48 mg/ml [geometric standard error of the mean (GSEM), 1.40] which was significantly (p<0.01) greater than that with the placebo [0.25 mg/ml (GSEM, 1.36)]. The value with theophylline [0.32 mg/ml (GSEM, 1.54)] did not differ significantly from that with cilostazol or the placebo. Baseline forced vital capacity (FVC) and FEV1 values significantly (p<0.05 and p<0.001, respectively) increased with cilostazol from 2.73+/-0.25 and 1.52+/-0.17 to 2.86+/-0.23 and 1.65+/-0.17 liters, respectively. Theophylline did not significantly increase the FVC or FEV1 value. CONCLUSION: It is suggested that the PDE3 inhibitor, cilostazol, has bronchodilator and bronchoprotective effects in elderly asthmatics. PMID- 9414145 TI - KF19514, a phosphodieterase 4 and 1 inhibitor, inhibits PAF-induced lung inflammatory responses by inhaled administration in guinea pigs. AB - Phosphodiesterase (PDE) 4 inhibitors are well known for their inhibitory effect on bronchoconstriction and inflammation and may be promising anti-asthma drugs. Platelet-activating factor (PAF) has been proposed as an inflammatory mediator to be relevant to asthma. It causes bronchoconstriction, airway microvascular leakage, inflammatory cell accumulation in the lung and hyperresponsiveness. In this study, we therefore have investigated the anti-asthmatic effects of the inhaled KF19514 [5-phenyl-3'-(3-pyridyl)methyl-3H-imidazo(4,5-c)(1,8) naphthyridin-4(5H)-one], a PDE 4 and 1 inhibitor, on PAF-induced lung inflammatory responses in guinea pigs. The inhaled KF19514 (0.0001-0.01%) significantly inhibited PAF-induced eosinophil and neutrophil accumulation into the airway and hyperresponsiveness in guinea pigs. The IC50 value of KF19514 against eosinophil accumulation was 14.8 microM (0.00063%). Moreover, the effect of KF19514 on the electrical field stimulation-induced bronchial contraction was examined using the main bronchi of guinea pigs in vitro. KF19514 inhibited both cholinergic and tachykininergic contraction and, in particular, produced a potent inhibitory effect on tachykininergic contraction (IC50 = 0.49 microM). The mechanism by which KF19514 inhibited the PAF-induced hyperresponsiveness may in part be the suppression of the tachykinin release. Based on these results, it was demonstrated that the inhaled KF19514 might have a significant potential effect on the inflammatory cell accumulation and hyperresponsiveness induced by PAF. PMID- 9414146 TI - Cold stimulation test and histamine release in primary acquired cold urticaria. AB - The patient was a 10-year-old boy who complained of urticaria upon exposure to cold air and after swimming in the pool. He also had seasonal asthma and wheezing after strenuous activities. To determine whether he had primary acquired cold urticaria, we performed a cold stimulation test twice. We likewise wanted to know whether a difference in response with regard to histamine release existed between blood samples taken from the challenged and the unchallenged sites. We obtained blood samples for histamine release initially at the site opposite the challenged forearm, and then on the same side on two separate occasions. We noted the appearance of constitutional signs and symptoms and correlated the time of their appearance with the result of histamine levels. The patient complained of pruritus and wheals appeared at the 5 minute in both tests. Results of plasma histamine release in the two measurements showed the highest releasability at 15 min. Our findings revealed that histamine is released systemically in response to cold stimulation regardless of the site where the blood sample was obtained. PMID- 9414147 TI - Toy-related injuries among children and teenagers--United States, 1996. AB - Each year, approximately two billion toys and games are sold in the United States. Although most toys are safe when risks are measured against the frequency of their use, children are at risk for some toy-related injuries and deaths. To characterize the magnitude of this problem, CDC analyzed data from the U.S. Consumer Product Safety Commission (CPSC) for 1996. This report summarizes this analysis and underscores the importance of parental participation in the selection and use of toys. PMID- 9414148 TI - Outbreak of staphylococcal food poisoning associated with precooked ham--Florida, 1997. AB - On September 27, 1997, a community hospital in northeastern Florida notified the St. Johns County Health Department about several persons who were treated in the emergency department because of gastrointestinal illnesses suspected of being associated with a common meal. This report summarizes the investigation of the outbreak by the Florida Department of Health; the findings implicated staphylococcal intoxication as the cause of illness among some persons who attended a retirement party on September 26, 1997. PMID- 9414149 TI - Update: influenza activity--United States, 1997-98 season. AB - In collaboration with the World Health Organization (WHO), its collaborating laboratories, and state and local health departments, CDC conducts surveillance to monitor influenza activity and to detect antigenic changes in the circulating strains of influenza viruses. This report summarizes influenza surveillance in the United States from September 1 through December 12, 1997, which indicates that influenza activity is at typical levels for this time of year and that influenza A(H3N2) viruses have been most commonly isolated. PMID- 9414150 TI - Poliomyelitis--United States, Canada. AB - As of May 22, an additional case of polio caused by type 1 poliovirus has been reported in Pennsylvania, bringing to 4 the total number of such cases this year. Two other states have reported suspected cases. Three of the confirmed and both suspected cases are in Amish residents. In addition, Ontario, Canada, has confirmed a case of paralytic poliomyelitis (type 1 virus) in an Amish woman. PMID- 9414151 TI - Follow-up on poliomyelitis--United States, Canada, Netherlands. 1979. PMID- 9414152 TI - Dental service use and dental insurance coverage--United States, Behavioral Risk Factor Surveillance System, 1995. AB - In the United States, 94% of adults have evidence of past or current tooth decay, and only one third of adults aged 35-44 years have all of their permanent teeth. Dental insurance is associated with increased use of dental services and improved oral health status. This report summarizes state-specific and aggregated state data on both private and public sources of dental insurance coverage and the use of dental services among adults in 25 states who participated in the oral health module of the 1995 Behavioral Risk Factor Surveillance System (BRFSS). The findings indicate that nearly half (44.3%) of adults in this survey reported having no dental insurance coverage. PMID- 9414153 TI - Isolation of avian influenza A(H5N1) viruses from humans--Hong Kong, May-December 1997. AB - A strain of influenza virus that previously was known to infect only birds has been associated with infection and illness in humans in Hong Kong. The first known human case of influenza type A(H5N1) occurred in a 3-year-old child who died from respiratory failure in May 1997. In Hong Kong, the virus initially was identified as influenza type A, but the subtype could not be determined using standard reagents. By August, CDC; the National Influenza Center, Rotterdam, the Netherlands; and the National Institute for Medical Research, London, United Kingdom, had independently identified the virus as influenza A(H5N1). An investigation conducted during August-September by the Hong Kong Department of Health and CDC excluded the possibility of laboratory contamination. Since this initial case was identified, six additional persons in Hong Kong have been confirmed to have influenza A(H5N1) infection, and two possible cases have been identified. This report summarizes the nine cases identified thus far and describes preliminary findings from the ongoing investigation, which indicate that multiple influenza A(H5N1) infections have occurred and that both the source and mode of transmission are uncertain at this time. PMID- 9414154 TI - National Institutes of Health sets up its own bioethics panel. PMID- 9414155 TI - Researcher sues over 'fraud' sanction. PMID- 9414156 TI - Inquiry looks into Indian cancer deaths. PMID- 9414157 TI - Plant cyclic AMP comes in from the cold. PMID- 9414158 TI - B lymphocytes and neuroinvasion. PMID- 9414159 TI - Regeneration of adult axons in white matter tracts of the central nervous system. AB - It is widely accepted that the adult mammalian central nervous system (CNS) is unable to regenerate axons. In addition to physical or molecular barriers presented by glial scarring at the lesion site, it has been suggested that the normal myelinated CNS environment contains potent growth inhibitors or lacks growth-promoting molecules. Here we investigate whether adult CNS white matter can support long-distance regeneration of adult axons in the absence of glial scarring, by using a microtransplantation technique that minimizes scarring to inject minute volumes of dissociated adult rat dorsal root ganglia directly into adult rat CNS pathways. This atraumatic injection procedure allowed considerable numbers of regenerating adult axons immediate access to the host glial terrain, where we found that they rapidly extended for long distances in white matter, eventually invading grey matter. Abortive regeneration correlated precisely with increased levels of proteoglycans within the extracellular matrix at the transplant interface, whereas successfully regenerating transplants were associated with minimal upregulation of these molecules. Our results demonstrate, to our knowledge for the first time, that reactive glial extracellular matrix at the lesion site is directly associated with failure of axon regrowth in vivo, and that adult myelinated white matter tracts beyond the glial scar can be highly permissive for regeneration. PMID- 9414160 TI - The cellular prion protein binds copper in vivo. AB - The normal cellular form of prion protein (PrPC) is a precursor to the pathogenic protease-resistant forms (PrPSc) believed to cause scrapie, bovine spongiform encephalopathy (BSE) and Creutzfeldt-Jakob disease. Its amino terminus contains the octapeptide PHGGGWGQ, which is repeated four times and is among the best preserved regions of mammalian PrPC. Here we show that the amino-terminal domain of PrPC exhibits five to six sites that bind copper (Cu(II)) presented as a glycine chelate. At neutral pH, binding occurs with positive cooperativity, with binding affinity compatible with estimates for extracellular, labile copper. Two lines of independently derived PrPC gene-ablated (Prnp0/0) mice exhibit severe reductions in the copper content of membrane-enriched brain extracts and similar reductions in synaptosomal and endosome-enriched subcellular fractions. Prnp0/0 mice also have altered cellular phenotypes, including a reduction in the activity of copper/zinc superoxide dismutase and altered electrophysiological responses in the presence of excess copper. These findings indicate that PrPC can exist in a Cu-metalloprotein form in vivo. PMID- 9414161 TI - A crucial role for B cells in neuroinvasive scrapie. AB - Although prion proteins are most efficiently propagated through intracerebral inoculation, peripheral administration has caused the diseases kuru, iatrogenic Creutzfeldt-Jakob disease (CJD), bovine spongiform encephalopathy (BSE) and new variant CJD. The development of neurological disease after peripheral inoculation depends on prion expansion within cells of the lymphoreticular system. Here we investigate the identity of these cells by using a panel of immune-deficient mice inoculated with prions intraperitoneally: we found that defects affecting only T lymphocytes had no apparent effect, but that all mutations that disrupted the differentiation and response of B lymphocytes prevented the development of clinical scrapie. As an absence of B cells and of antibodies correlates with severe defects in follicular dendritic cells, a lack of any of these three components may prevent the development of clinical scrapie. However, we found that scrapie developed after peripheral inoculation in mice expressing immunoglobulins that were exclusively of the M subclass and without detectable specificity for the normal form of the prion PrPC, and in mice which had differentiated B cells but no functional follicular dendritic cells. We conclude that differentiated B cells are crucial for neuroinvasion by scrapie, regardless of the specificity of their receptors. PMID- 9414162 TI - Neuregulin-beta induces expression of an NMDA-receptor subunit. AB - Neuregulins (also known as ARIA, NDF, heregulin, GGF) are a family of widely expressed growth and differentiation factors. Neuregulins secreted from motor neurons accumulate at maturing neuromuscular junctions, where they stimulate transcription of genes encoding specific acetylcholine receptors. How these factors function at central synapses, however, is unknown. In the maturing cerebellum, neuregulins are concentrated in glutamatergic mossy fibres that innervate granule cells in the internal granule-cell layer. We have analysed the effects of neuregulins on the expression of genes encoding NMDA (N-methyl-D aspartate) receptors in the cerebellum, because receptor composition changes dramatically as expression of the receptor NR2C subunit is specifically induced in neurons in the internal granule-cell layer during synaptogenesis. Here we report that addition of a neuregulin-beta isoform to cultured cerebellar slices specifically increases the expression of NR2C messenger RNAs by at least 100 fold; effects are only minor with a neuregulin-alpha isoform. This stimulation of NR2C expression requires synaptic activity by NMDA receptors, as well as neuregulin-beta. Addition of the NMDA-receptor-channel blocker AP-5 prevents upregulation of the NR2C subunit by neuregulin, whereas an AMPA/kainate-receptor antagonist does not. Consistent with these effects of neuregulin, we find that granule cells express its receptors ErbB2 and ErbB4 before the NR2C subunit of the NMDA receptor. Our results indicate that neuregulins regulate the composition of neurotransmitter receptors in maturing synapses in the brain, in a manner analogous to the neuromuscular junction. PMID- 9414163 TI - Cyclic-nucleotide-gated channels mediate synaptic feedback by nitric oxide. AB - Cyclic-nucleotide-gated (CNG) channels in outer segments of vertebrate photoreceptors generate electrical signals in response to changes in cyclic GMP concentration during phototransduction. CNG channels also allow the influx of Ca2+, which is essential for photoreceptor adaptation. In cone photoreceptors, cGMP triggers an increase in membrane capacitance indicative of exocytosis, suggesting that CNG channels are also involved in synaptic function. Here we examine whether CNG channels reside in cone terminals and whether they regulate neurotransmitter release, specifically in response to nitric oxide (NO), a retrograde transmitter that increases cGMP synthesis and potentiates synaptic transmission in the brain. Using intact retina, we show that endogenous NO modulates synapses between cones and horizontal cells. In experiments on isolated cones, we show directly that CNG channels occur in clusters and are indirectly activated by S-nitrosocysteine (SNC), an NO donor. Furthermore, both SNC and pCPT cGMP, a membrane-permeant analogue of cGMP, trigger the release of transmitter from the cone terminals. The NO-induced transmitter release is suppressed by guanylate cyclase inhibitors and prevented by direct activation of CNG channels, indicating that their activation is required for NO to elicit release. These results expand our view of CNG channel function to include the regulation of synaptic transmission and mediation of the presynaptic effects of NO. PMID- 9414165 TI - Kaposi's sarcoma and Kaposi's sarcoma associated herpesvirus (human herpesvirus 8): where are we now? PMID- 9414164 TI - Identification and role of adenylyl cyclase in auxin signalling in higher plants. AB - Cyclic AMP is an important signalling molecule in prokaryotes and eukaryotes, but its significance in higher plants has been generally doubted because they have low adenylyl cyclase activity and barely detectable amounts of cAMP. Here we used activation T-DNA tagging to create tobacco cell lines that can proliferate in the absence of the phytohormone auxin in the culture media. The sequence tagged in one line, axi 141, was used to isolate a complementary DNA encoding adenylyl cyclase, the first from a higher plant. Sequence analysis reveals that the tobacco adenylyl cyclase is probably soluble, contains characteristic leucine rich repeats, and bears similarity with adenylyl cyclase from the yeast Schizosaccharomyces pombe. Expression of the cDNA in Escherichia coli results in an increase in endogenous cAMP levels, and in yeast its expression functionally complements the cry1 mutation. Tobacco protoplasts treated with cAMP, or the adenylyl cyclase activator forskolin, no longer require auxin to divide. This finding, together with the observation that the adenylyl cyclase inhibitor dideoxyadenosine inhibits cell proliferation in the presence of auxin, suggests that cAMP is involved in auxin-triggered cell division in higher plants. PMID- 9414166 TI - Estonian cancer registry straddles two worlds. PMID- 9414167 TI - Finnish registry: more than 50 years experience and still counting. PMID- 9414168 TI - End-of-life suffering begins to attract research attention. PMID- 9414169 TI - Presidential bioethics subcommittee wrestles with confidentiality of tissue donors. PMID- 9414170 TI - New techniques catch lung cancers earlier. PMID- 9414171 TI - Medical research is a public priority, polls show. PMID- 9414172 TI - Hepatocellular carcinoma: from gene to public health. AB - Liver diseases associated with chronic hepatitis B virus (HBV) infection, including hepatocellular carcinoma, account for more than 1 million deaths annually worldwide. In addition to HBV infection, other risk factors are involved in the etiology of hepatocellular carcinoma and, among these, dietary exposure to the carcinogenic aflatoxins is of particular importance in certain regions of southeast Asia and sub-Saharan Africa. The relative contributions of these two risk factors and the mechanism of the interaction between them in the pathogenesis of hepatocellular carcinoma are still poorly understood. The recently developed individual biochemical and molecular markers of aflatoxin exposure, i.e., aflatoxin-albumin adducts in blood and a specific GC to TA transversion mutation in codon 249 of the p53 gene (249ser p53 mutation) in hepatocellular carcinomas, permit a better quantitative estimation of aflatoxin exposure in different populations of the world. A comprehensive summary of the data from our laboratory and the literature, based on a large number (>1000) of individual cases of hepatocellular carcinoma, is presented here and shows the following: 1) A high level and high prevalence of exposure to aflatoxins occur in West Africa, Mozambique, and some regions of China; 2) a high prevalence of the 249ser p53 mutation is detected in these countries; and 3) hepatocellular carcinomas from countries with low or no exposure to aflatoxins show a very low prevalence of the 249ser p53 mutation and distinctly different p53 mutation spectra, probably indicating different etiologies. Experimental and epidemiologic studies demonstrate an interaction between HBV infection and aflatoxins in hepatocarcinogenesis. The relevance of the biochemical/molecular markers of aflatoxin exposure, HBV vaccination, and the reduction of aflatoxin exposure, in addition to the interaction between HBV infection and other risk factors in liver carcinogenesis, are discussed with regard to the implementation of measures for primary prevention. PMID- 9414173 TI - Tobacco addiction: implications for treatment and cancer prevention. AB - The American Society of Clinical Oncology and the National Cancer Institute convened a symposium in June 1996 on tobacco addiction. Additional support for the symposium was provided by the American Medical Women's Association and the American Society of Preventive Oncology. The goals of this conference were to describe the burden and public health consequences of tobacco addiction, to describe the state of science for the treatment of nicotine dependence, and to explore new strategies to increase quit rates and to prevent the uptake of tobacco use. This article summarizes and integrates the meeting presentations on tobacco addiction and includes the topics of smoking prevalence; psychobiologic aspects of nicotine dependence; and implications for disease, treatment, and prevention. Comments on regulatory approaches and national strategies for reducing dependence are also summarized in presentations by Dr. David Kessler, former Food and Drug Administration Commissioner, and Dr. C. Everett Koop, former U.S. Surgeon General. PMID- 9414174 TI - Expression of human herpesvirus 8-encoded cyclin D in Kaposi's sarcoma spindle cells. AB - BACKGROUND: Human herpesvirus 8 (HHV-8) DNA sequences have been detected in Kaposi's sarcoma, in primary effusion lymphoma (an unusual high-grade non Hodgkin's lymphoma seen primarily in patients with acquired immunodeficiency syndrome [AIDS]), and in Castleman's disease (a rare lymphoproliferative disorder); however, proof that HHV-8 is involved in the pathogenesis of these diseases remains to be established. HHV-8 contains a gene, i.e., v-cyclin D, that is a homologue of the cellular cyclin D2 gene, which encodes a protein that promotes passage through G1 phase of the cell cycle. Previous studies have identified v-cyclin D messenger RNA (mRNA) in biopsy specimens of Kaposi's sarcoma. In this study, we isolated a full-length v-cyclin D complementary DNA and characterized the pattern of v-cyclin D mRNA expression in Kaposi's sarcoma. METHODS: Standard methods were used to construct and to screen HHV-8 genomic and complementary DNA libraries. Reverse transcription-polymerase chain reaction (RT PCR) methods and in situ hybridization with RNA probes were used to examine v cyclin D mRNA expression. RESULTS: RT-PCR demonstrated the presence of v-cyclin D mRNA in biopsy specimens of AIDS-related Kaposi's sarcoma, in early-passage spindle cells from classical (i.e., not AIDS-related) Kaposi's sarcoma, and in spindle cells isolated from the peripheral blood of patients with AIDS-related Kaposi's sarcoma. In situ hybridization indicated that mRNAs for v-cyclin D and kaposin, an HHV-8 latency-associated gene, were present in approximately 1% of the spindle cells in early patch lesions and approximately 60% of the spindle cells in late nodular lesions of Kaposi's sarcoma. CONCLUSIONS: Spindle cells of Kaposi's sarcoma, which have been regarded as the tumor cells of this cancer, contain v-cyclin D mRNA. Expression of v-cyclin D protein may be involved in the pathogenesis of Kaposi's sarcoma by promoting cell proliferation. PMID- 9414175 TI - Telomerase activity and survival of patients with node-positive breast cancer. AB - BACKGROUND: Shortening of telomeres (specialized structures at the ends of chromosomes) beyond a certain length may signal a cell to stop dividing and to enter senescence. A ribonucleoprotein enzyme, telomerase, is a key component in maintaining telomere length. Because the majority of cancers express telomerase but most normal somatic tissues do not, we measured the level of telomerase expression in primary breast cancer specimens for correlation with traditional prognostic indicators and disease outcome. METHODS: Telomerase activity was measured in frozen human breast cancer specimens by use of the Telomeric Repeat Amplification Protocol (TRAP) assay. The level of telomerase activity was expressed as total product generated (TPG) and was corrected for specimen cellularity by expressing it as a ratio of TPG to the sample's 28S ribosomal RNA content. RESULTS: A preliminary study of 150 breast cancer specimens demonstrated that telomerase activity correlated with the fraction of cells in S phase of the cell cycle (r(sp) = .23). In a larger prognostic study of 398 tumors from patients with lymph node-positive breast cancer, telomerase expression correlated with S-phase fraction, progesterone receptor level, DNA ploidy, and lymph node status. After correcting for sample cellularity, increasing TPG levels were associated with decreased disease-free survival (P = .041) and overall survival (P = .009) of the patients. The telomerase activity level remained strongly predictive of death (P = .027) and marginally predictive of disease recurrence (P = .08) after adjustment for other prognostic factors. All P values are two-sided. CONCLUSIONS: Telomerase activity in human breast cancers is associated with a more aggressive tumor phenotype in patients. PMID- 9414176 TI - Green tea constituent epigallocatechin-3-gallate and induction of apoptosis and cell cycle arrest in human carcinoma cells. AB - BACKGROUND AND PURPOSE: The polyphenolic compounds present in green tea show cancer chemopreventive effects in many animal tumor models. Epidemiologic studies have also suggested that green tea consumption might be effective in the prevention of certain human cancers. We investigated the effect of green tea polyphenols and the major constituent, epigallocatechin-3-gallate, on the induction of apoptosis (programmed cell death) and regulation of cell cycle in human and mouse carcinoma cells. METHODS: Human epidermoid carcinoma cells (cell line A431), human carcinoma keratinocyte (cell line HaCaT), human prostate carcinoma cells (cell line DU145), mouse lymphoma cells (cell line L5178Y), and normal human epidermal keratinocytes (NHEKs) were used. Apoptosis was assessed by 1) the formation of internucleosomal DNA fragments by agarose gel electrophoresis, 2) confocal microscopy, and 3) flow cytometry after tagging the DNA fragments by fluorescence label. The distribution of cells in different phases of the cell cycle was analyzed by flow cytometry. RESULTS: Treatment of A431 cells with green tea polyphenols and its components, epigallocatechin-3 gallate, epigallocatechin, and epicatechin-3-gallate, resulted in the formation of internucleosomal DNA fragments, characteristic of apoptosis. Treatment with epigallocatechin-3-gallate also resulted in apoptosis in HaCaT, L5178Y, and DU145 cells, but not in NHEK. Confocal microscopy and flow cytometry confirmed the findings. The DNA cell cycle analysis showed that in A431 cells, epigallocatechin 3-gallate treatment resulted in arrest in the G0-G1 phase of the cell cycle and a dose-dependent apoptosis. CONCLUSIONS: Green tea may protect against cancer by causing cell cycle arrest and inducing apoptosis. It needs to be evaluated in human trials. PMID- 9414177 TI - Overexpression of C/EBPbeta-LIP, a naturally occurring, dominant-negative transcription factor, in human breast cancer. AB - BACKGROUND: When cells fail to maintain a balance between proliferation, terminal differentiation, and programmed cell death, cancer often results. The CCAAT/enhancer-binding protein (C/EBP) family of transcription factors regulates many genes involved in the processes of proliferation and terminal differentiation. The messenger RNA for C/EBPbeta is translated into two major isoforms, LAP (liver-enriched activating protein) and LIP (liver-enriched inhibitory protein). LIP levels appear to be elevated in mouse mammary tumors but not in hyperplastic mammary tissues. We tested whether LIP expression is elevated in human breast cancer and whether elevated expression is associated with biologic predictors of the aggressiveness of the disease. METHODS: Homogenates of infiltrating ductal carcinoma specimens from 39 women were analyzed for C/EBPbeta protein content by western blot analysis, and the ratio of LAP to LIP in specimens containing high levels of LIP (i.e., levels approximately 15 times higher than those in tumor specimens that express little or no LIP) was also determined. Nonparametric statistical analyses were performed. RESULTS: LIP was present at high levels in nine of 39 specimens of infiltrating ductal carcinoma. Eight of the nine specimens of infiltrating ductal carcinoma that contained high levels of LIP were negative for estrogen receptor and progesterone receptor (ER /PR-); all nine tumors were aneuploid and poorly differentiated, and eight of nine were highly proliferative. Of the tumors that contained LIP at low or nondetectable levels, six of 30 were ER-/PR-, 17 of 29 were aneuploid, eight of 27 were highly proliferative, and 11 of 30 were poorly differentiated. IMPLICATION: LIP expression should be evaluated further as a prognostic marker for patients with breast cancer. PMID- 9414178 TI - Re: five-day oral etoposide treatment for advanced small-cell lung cancer: randomized comparison with intravenous chemotherapy. PMID- 9414179 TI - Re: prostate cancer susceptibility locus on chromosome 1q: a confirmatory study. PMID- 9414180 TI - Re: Researchers make slow headway in managing dry mouth. PMID- 9414181 TI - Gemcitabine-induced hemolytic uremic syndrome: a case report. PMID- 9414182 TI - Immunophenotyping of malignant mesothelioma. PMID- 9414183 TI - The value of antibodies 44-3A6, SM3, HBME-1, and thrombomodulin in differentiating epithelial pleural mesothelioma from lung adenocarcinoma: a comparative study with other commonly used antibodies. AB - The distinction between pleural mesothelioma and peripheral pulmonary adenocarcinoma involving the pleura continues to be a diagnostic problem in surgical pathology. In recent years, the use of various immunohistochemical markers to facilitate this differential diagnosis has become common. In this study, the value of monoclonal antibodies 44-3A6, SM3, HBME-1, and thrombomodulin is compared in the differentiation of these conditions. Fifteen (68.2%) of 22, and 10 (52.6%) of 19 mesotheliomas stained positively with 44-3A6 and SM3, respectively, whereas all 23 (100%) adenocarcinomas reacted with both antibodies. Sixteen (80%) of 20 mesotheliomas and 14 (63.6%) of 22 lung adenocarcinomas reacted with HBME-1, whereas 16 (80%) of 20 mesotheliomas and only three (11.1%) of 27 adenocarcinomas were positive for thrombomodulin. Because thrombomodulin was expressed in most mesotheliomas but in only a few lung adenocarcinomas, this marker may have some diagnostic value when it is included in the standard immunohistochemical panel of markers used in the evaluation of mesotheliomas, especially when a positive marker for mesothelioma is needed. Antibodies 44-3A6, SM3, and HBME-1 have no practical value in discriminating epithelial pleural mesothelioma from lung adenocarcinoma. PMID- 9414185 TI - Anaplastic large cell lymphoma: a distinct molecular pathologic entity: a reappraisal with special reference to p80(NPM/ALK) expression. AB - The p80(NPM/ALK) expression activated by the t(2;5) (p23;q35) translocation recently has been shown to play an important role in the pathogenesis of anaplastic large cell lymphoma (ALCL). However, the clinicopathologic significance of identification of p80 among ALCL cases has not been completely resolved. Difficulties also exist in the histologic and immunophenotypic identification of ALCL and Hodgkin's disease (HD) as separate processes, often complicating the clinicopathologic evaluation of and therapeutic approach to these entities. In order to clarify these issues, 67 specimens of ALCL and 63 specimens of HD (31 of the nodular-sclerosing type [NS-HD] and 32 of the mixed cellularity type [MC-HD]) were immunostained using anti-p80 antibody and other relevant markers on paraffin sections. The clinicopathologic and immunophenotypic features were reviewed on the basis of p80 reactivity. The expression of p80 was detected in 43 of 67 cases of ALCL (64%), but none of HD. The p80+ ALCL cases constituted a very homogeneous group of tumors, characterized by the occurrence in a much younger group and relatively more favorable clinical course than the p80- ALCL, which were in keeping with the data previously reported. They showed virtually the identical immunophenotypic findings of p80+, CD30+, EMA+, CD15-, bcl-2-, and Epstein-Barr virus (EBV) with T- and null-cell phenotype, and showed the distinct morphologic features, including three cases of lymphohistiocytic/small-cell variant, as follows: the indented nuclei, often termed as reniform, embryolike, and horseshoelike; multiple, irregular, but indistinct nucleoli; and few reactive cells of eosinophils and epithelioid cells. Conversely, the 24 p80- ALCL cases, in which epithelial membrane antigen (EMA) and bcl-2 positivities were 33% and 55%, respectively, were heterogeneous and could be subdivided into five different categories, namely (a) 11 cases of HD like ALCLs, (b) six cases of p80 common ALCL, (c) three cases of secondary ALCL, (d) two cases of primary cutaneous ALCL, and (e) two cases of primary classical ALCL that lacked p80 expression. This study clearly demonstrated that the immunohistochemical detection of p80 is of a crucial importance in delineating the biologically distinct entity of "primary classical ALCL" from various diseases that show morphologic and immunohistologic overlap, including HD and HD like ALCL. PMID- 9414184 TI - The immunohistochemical diagnostic panel for epithelial mesothelioma: a reevaluation after heat-induced epitope retrieval. AB - The immunohistochemical diagnosis between epithelial mesothelioma and adenocarcinoma is currently based on the use of a panel of antibodies to adenocarcinoma-associated antigens and a few antibodies to mesothelial-associated antigens. Since the introduction of epitope retrieval methods, the sensitivity of many antibodies has been enhanced. Thus, a reevaluation of the mesothelioma/adenocarcinoma diagnostic panel becomes necessary. We studied 268 paraffin-embedded formalin-fixed tumor samples that included 57 epithelial mesotheliomas and 211 adenocarcinomas of various origins, comparing an extensive antibody panel with and without heat-induced epitope retrieval (HIER). Marked increase in the sensitivity of several antibodies, with no loss of specificity, was found when HIER was used. After statistical analysis, the antibodies to the epithelial glycoproteins carcinoembryonic antigen, BerEp4, and Bg8 emerged as the best discriminators between adenocarcinoma and epithelial mesothelioma within the entire panel. The mesothelium-associated antibodies, HBME-1, calretinin, and thrombomodulin were less sensitive and less specific than the former, although they were found to be useful on certain cases. Antibodies to cytokeratins and vimentin, although of minor diagnostic value in this context, may be helpful to evaluate the quality of antigen preservation. This study confirms the value of immunohistochemistry to accurately distinguish mesothelioma from adenocarcinoma when an antibody panel approach is used. The addition of heat-induced epitope retrieval methods increases the effectiveness of the procedure and is recommended for most of the antibody panel members. PMID- 9414186 TI - Sclerosing perineurioma: a clinicopathologic study of 19 cases of a distinctive soft tissue lesion with a predilection for the fingers and palms of young adults. AB - This report describes 19 cases of a distinctive sclerosing perineurial tumor of the hands. Fourteen patients were male and five were female (age range 9-55 years; median age 24.5 years). The process typically presented as a painless mass and was present from 6 months to 40 years before resection. Sites of involvement were the thumb (n = 6); index (n = 3), middle (n = 4), and ring (n = 4) fingers; and the palm (n = 2). The lesions were generally well marginated but nonencapsulated. They had a firm, fibrous consistency and ranged in size from 0.7 to 3.3 cm in maximum dimension. Microscopic examination showed abundant dense collagen and variable numbers of small, epithelioid, and spindled cells exhibiting corded, trabecular, and whorled (onion bulblike) growth patterns. Immunoreactivity was present for epithelial membrane antigen (15 of 15); a cytokeratin cocktail containing AE1, AE3, and CK1 (four of 14); CAM 5.2 (one of 12); vimentin (12 of 12); muscle-specific actin (nine of 14); alpha-smooth muscle actin (six of 14); collagen IV (six of six); laminin (five of six); and CD99 (three of five). Ultrastructural features consistent with perineurial cells were noted. All of the lesions were locally excised. Follow-up was obtained for seven patients, with mean and median follow-up intervals of 12 years 7 months and 10 years 6 months, respectively. None of the lesions have recurred. This study advances the morphologic spectrum of perineurioma, a rare tumor of nerve sheath derivation. Familiarity with this distinctive subtype should help to avoid confusion with other processes, including a fibroma of tendon sheath, the sclerotic fibroma associated with Cowden's disease, an epithelioid neurofibroma, a late stage of tenosynovial giant cell tumor, and sclerosing adnexal tumors. PMID- 9414187 TI - Neurofibroma and cellular neurofibroma with atypia: a report of 14 tumors. AB - The clinical significance of nuclear atypia in neurofibromas that lack necrosis or significant mitotic activity has not been systematically studied. We reviewed 14 neurofibromas from six patients with mild to marked nuclear atypia, with low mitotic activity in some tumors. Five tumors also had areas of increased cellularity consistent with cellular neurofibroma. Necrosis was absent. All patients were treated by conservative excision. Clinical follow-up, ranging from 8 months to 6 years, showed that none of the tumors recurred or metastasized. To further characterize these neoplasms, we assessed p53 expression, proliferation rate, and DNA content because these methods have been suggested by others as useful in differentiating benign from malignant nerve sheath tumors. p53 expression was detected by immunostaining in one tumor with 5% positive cells and in two tumors with rare positive cells (<1%). The remaining 11 tumors were negative. Tumor cell proliferation rate as determined by Ki-67 immunostaining showed <5% positive cells in 13 tumors. In one tumor, 10% of the cells were Ki-67 positive. Using flow cytometry methods and paraffin-embedded tissue, all tumors had diploid DNA content with an S phase fraction ranging from 5.2% to 18.2% (mean 9.4%). No significant differences were observed between the neurofibromas and cellular neurofibromas. For comparison, we studied three malignant peripheral nerve sheath tumors (MPNSTs). All MPNSTs had relatively high p53 (range 10-16%; mean 12%) and Ki-67 (range 32-42%; mean 38.0%) staining. One of the MPNSTs was aneuploid. The S phase fraction of the MPNSTs ranged from 8.1% to 51.8% (mean 28.6%). These results suggest that clinically benign neurofibromas, both usual and cellular types, can have significant cytologic atypia that can be accompanied by low mitotic activity. Conservative surgical excision for these tumors is adequate. The results of p53 and Ki-67 immunostaining and DNA content and S-phase analysis by flow cytometry support this interpretation. In addition, in tumors with borderline histologic findings, results of these ancillary studies may be useful in distinguishing benign from malignant nerve sheath tumors. PMID- 9414188 TI - Differential expression of the intermediate filament peripherin in cutaneous neural lesions and neurotized melanocytic nevi. AB - Peripherin is an intermediate filament involved in growth and development of the peripheral nervous system, and is produced by neurons and the beta cells of the islets of Langerhans. Recently, malignant melanomas and some melanocytic nevi have been shown to express peripherin. It is unknown if Schwann cells, also derived from the neural crest, express peripherin. Expression of peripherin was evaluated by immunohistochemistry in cutaneous lesions characterized by a prominent Schwann cell component including 26 neurofibromas (NF), 10 schwannomas (SCH), seven granular cell tumors, and five palisaded encapsulated neuromas (PEN); 13 neurotized melanocytic nevi (NMN) also were evaluated because these lesions contain Wagner-Meissnerlike structures and type C nevus cells, which exhibit a "schwannian" phenotype. Peripherin was detected in the axons of normal peripheral nerves. NF and PEN contained numerous axons dispersed throughout the lesions, whereas only scattered small nerves were seen in GCT. In SCH, only rare axons were labeled, mostly at the periphery of the lesions. All other cells in these four types of lesions, therefore including Schwann cells, were not labeled. In most NMN, labeled axons were identified within the lesions. In a few cases, rare epithelioid melanocytes within the superficial portions of the nevi were labeled. The Wagner-Meissnerlike structures and type C nevus cells (schwannian) were not labeled in any lesion; however, numerous labeled axons invested these areas. Because there are different relative numbers of peripherin-labeled axons throughout NF, PEN, some nevi, and SCH, analysis of peripherin expression may be helpful in the diagnosis of these lesions. Neurons and some epithelioid melanocytes, in contrast to type C nevus cells and Schwann cells of NF and SCH, express peripherin, providing further evidence for a transition from a more neuronal to a more schwannian phenotype during the normal maturation sequence of melanocytes in nevi. PMID- 9414189 TI - Meningioma grading: an analysis of histologic parameters. AB - Histologic grading of meningiomas has prognostic and sometimes therapeutic implications, but diagnostic criteria for atypical meningioma are vague, and the significance of brain invasion in the determination of malignancy remains controversial. We reviewed our experience with 581 patients whose meningiomas were resected at Mayo Clinic during the years 1978 through 1988. All patients were followed until death or a median of 9.0 years. Ten histologic parameters were assessed and compared with recurrence-free survival. On univariate analysis, six variables were associated with recurrence, although most were statistically significant only in the subset of patients having undergone gross total tumor resection. On multivariate analyses, the most significant parameters were histologic brain invasion (when assessable) and maximal mitotic rate of at least four per 10 high-power fields (HPF). Also significant were combinations of at least three of the following four parameters: hypercellularity, architectural sheeting, macronucleoli, and small cell formation. Proposed grading criteria based on these findings yielded 81% classic, 15% atypical, and 4% brain invasive meningiomas with respective 5-year recurrence rates of 12%, 41%, and 56%. There was no association between histologic grade and either extent of surgical resection or patient age. However, male sex was associated with high-grade (atypical/brain invasive) tumors. Too few frankly anaplastic meningiomas were encountered for statistical analysis. Brain invasion and an increased mitotic index (at least four per 10 HPF) are the most powerful histologic factors prognostic for recurrence in meningiomas. We propose an objective definition for atypical meningioma based on our data. Because the difference in recurrence rates for brain invasive and atypical meningiomas was not statistically significant, it could not be determined whether brain invasion alone warrants a designation of malignancy. Likewise, we were unable to determine what constitutes histologic anaplasia due to the rarity of such cases. PMID- 9414190 TI - Insular carcinoma: a distinct de novo entity among follicular carcinomas of the thyroid gland. AB - We reclassified 720 nonmedullary invasive thyroid carcinomas diagnosed and treated between 1975 and 1993. Twenty-seven cases met the criteria of insular carcinoma and 29 cases those of widely invasive follicular carcinoma. Comparison of these histotypes with respect to pathologic stage and overall, relative, and visceral metastasis-free survival showed a significant association between histotype and pT and pN categories. In particular, pT4 (p < 0.001) and pN1 (p < 0.001) categories were more frequent in the insular carcinoma histotype. By contrast, no significant differences in overall, relative, or visceral metastasis free survival were observed between insular carcinoma and widely invasive follicular carcinoma. Molecular analysis by polymerase chain reaction-single strand conformation polymorphism demonstrated RAS gene family point mutations in five of eight cases analyzed in each of the two histotypes, with a high proportion of CAA-->AAA transversion at codon 61 of the N-RAS gene in insular carcinoma. These findings suggest that insular carcinoma represents a de novo entity distinct from widely invasive follicular carcinoma, that widely invasive follicular carcinoma has biologic characteristics more consistent with poorly differentiated than well-differentiated carcinomas, and that both insular carcinoma and widely invasive follicular carcinoma share similar molecular alterations. PMID- 9414191 TI - Adenocarcinoma of the epididymis: a report of four cases and review of the literature. AB - Primary epididymal carcinoma is extremely rare and has been the subject of only sporadic case reports; the validity of some of the reported tumors is questionable. We report our experience with four examples, which arose in men 27, 66, 77, and 81 years of age. All of them presented with scrotal masses; three of them had small hydroceles. None had a history of von Hippel-Lindau disease. Grossly, all of the tumors were centered in the epididymis and two were confined to it. One tumor invaded the periepididymal soft tissue and spermatic cord and the fourth invaded the adjacent testis. They were 2.0-7.0 cm in greatest diameter; three had foci of hemorrhage and necrosis. Microscopically, all of the tumors were adenocarcinomas. Two of them were composed of approximately equal numbers of simple tubules and more complex tubulopapillary formations lined by cuboidal or columnar predominantly clear cells that infiltrated the epididymal smooth muscle wall, periepididymal soft tissue, or both. The other two had large cysts into which grew complex papillary or confluent, back-to-back glands lined by columnar cells with clear, lightly amphophilic or eosinophilic cytoplasm. Necrosis was present in three cases. An undifferentiated, sheetlike growth of anaplastic tumor cells was present focally in one case. Cilia were absent. Small amounts of cytoplasmic glycogen were present in the two cases in which periodic acid-Schiff staining was performed. Immunohistochemical staining, performed in one case, demonstrated strong positivity for cytokeratins (AE1/3, Cam 5.2) and epithelial membrane antigen (luminal only). Results of staining for carcinoembryonic antigen, Leu M1, B72.3, and Ber-EP4 were negative. Ultrastructural analysis, performed in one case, showed well-developed desmosomal junctions, cytoplasmic multivesicular bodies, glycogen particles, and well developed cilia. Two of three patients with follow-up data died of disease 8 months and 2.5 years after diagnosis; the third was disease free 30 years after the diagnosis despite having paraaortic lymph node metastases at the time of presentation. After reviewing the literature and analyzing our cases, we conclude that bona fide examples of epididymal adenocarcinoma are usually tubular, tubulocystic, or tubulopapillary adenocarcinomas, often with an appreciable content of clear cells, which can usually be readily separated from other paratesticular malignant tumors, such as malignant mesothelioma and carcinomas of the mullerian type. Distinction from metastasis may be difficult and may depend largely on careful clinical evaluation. PMID- 9414192 TI - Hyalinizing spindle cell tumor with giant rosettes: a distinctive tumor closely resembling low-grade fibromyxoid sarcoma. AB - We report the findings of 19 cases of a previously undescribed spindle cell tumor of soft tissues that resembles a low-grade fibromyxoid sarcoma but contains distinctive rosettelike structures. The tumors occurred principally as a painless, slowly growing, deeply situated mass of the proximal extremities in young to middle-aged adults (age range 14-65 years; mean 38). Although grossly circumscribed, the tumors had infiltrative borders microscopically and were composed of bland spindled cells situated in a hyalinized to myxoid stroma. The most characteristic feature of the tumor was scattered large rosettelike structures that often merged with serpinginous areas of dense hyalinization. The rosettes consisted of a central collagen core surrounded by a rim of rounded cells morphologically and immunophenotypically different from the cells of the spindled stroma. These cells expressed a number of antigens, including S-100 protein, neuron-specific enolase, and leu 22, in contrast to the stroma, which usually lacked these antigens. Of the 12 patients with available follow-up information, one patient treated with simple excision clinically developed local recurrence of the tumor 20 months after initial biopsy. No other recurrences were reported during the limited follow-up period, and no patient developed metastatic disease. However, the favorable prognosis of the patients in our series to date may relate to the limited follow-up period (approximately 3 years), as well as initial treatment by wide excision in nearly half of the patients. We regard the hyalinizing spindle cell tumor with giant rosettes as a distinctive type of low grade fibroblastic tumor that with time may prove to behave similar to a low grade fibromyxoid sarcoma and, hence, to represent an unusual variant thereof. PMID- 9414193 TI - Atypical small acinar proliferation suspicious for malignancy in prostate needle biopsies: clinical significance in 33 cases. AB - Prostate needle biopsies occasionally contain an atypical small acinar proliferation (ASAP) that is suspicious for, but not diagnostic of, adenocarcinoma. The histologic features and clinical significance of this finding are unexplored. We evaluated 33 cases of ASAP with at least one follow-up needle biopsy seen at Mayo Clinic from 1993 to 1996. Numerous histologic and clinical features were assessed to determine their predictive value for adenocarcinoma on subsequent biopsy. Mean patient age was 61.6 years (range 45-72). Adenocarcinoma was identified on follow-up biopsy in 15 of 33 patients (45%), with a median follow-up of 9 months (range 1-27). Gleason score varied from 4 to 7 (mean 5.9). Two patients (6%) had subsequent diagnoses of ASAP after one and three repeat biopsies. Digital rectal examination, serum prostate-specific antigen, and a variety of histologic findings were not predictive of cancer on follow-up biopsy. These histologic findings included number of biopsy cores (mean 5.5), number of acini per focus of ASAP (mean 7.9), number of foci (mean one), variation in acinar size, nuclear enlargement (none, 12% of cases; mild, 45%; moderate, 33%; severe, 10%), nucleolar enlargement (none, 27%; mild, 46%; moderate, 27%), luminal mucin (39%), crystalloids (6%), focal chronic inflammation (64%), adjacent atrophy (100%), and adjacent high-grade prostatic intraepithelial neoplasia (PIN) (42%). Stratification of suspicion in cases of ASAP without PIN into three categories ("favor benign, uncertain, and favor carcinoma") was somewhat predictive of subsequent cancer (20%, 25%, and 60% of cases with subsequent cancer, respectively), but the results were not significant. The high predictive value of ASAP for subsequent adenocarcinoma warrants repeat biopsy. No single clinical or pathologic feature appeared to increase the likelihood of subsequent cancer. PMID- 9414194 TI - Prediction of extraprostatic extension of prostate cancer based on needle biopsy findings: perineural invasion lacks significance on multivariate analysis. AB - Extraprostatic extension (EPE) and seminal vesicle invasion (SVI) are adverse prognostic factors in prostate cancer, and their prediction before prostatectomy would be useful. Perineural invasion in needle biopsy has been advocated as a marker of extraprostatic extension, but its independent value as a predictor of stage has not been established. We studied 349 previously untreated men with prostatic adenocarcinoma who underwent bilateral pelvic lymphadenectomy and radical retropubic prostatectomy. All patients were clinically free of metastases and had cancer that was diagnosed on needle biopsy. Five preoperative variables were collected: clinical stage (TNM staging system), serum prostate-specific antigen (PSA), Gleason score on needle biopsy, presence or absence of perineural invasion, and proportion of the biopsy involved by cancer. The subsequent prostatectomy specimens were completely embedded, and whole mount sections were used to evaluate four outcome staging variables: EPE (absent/present), EPE (absent/unilateral/bilateral), seminal vesicle invasion, and pathologic stage (TNM). On univariate analysis, each preoperative variable was significantly associated with each outcome variable except for a lack of association between clinical stage and SVI. Perineural invasion in the biopsy predicted EPE with a sensitivity of 51%, specificity of 70%, positive predictive value of 49%, and negative predictive value of 71%. On multivariate analysis (stepwise logistic regression), only preoperative PSA, proportion of the biopsy involved by cancer, and Gleason score were significant (p < 0.05); perineural invasion and clinical stage had no independent predictive value for any of the outcome variables. We conclude that the finding of perineural invasion in needle biopsy of prostatic carcinoma has no independent predictive value for the presence of extraprostatic extension, seminal vesicle involvement, or pathologic stage in the radical prostatectomy. Accordingly, we no longer routinely evaluate this finding in biopsy specimens. PMID- 9414195 TI - Endocrine mucin-producing sweat gland carcinoma: a cutaneous neoplasm analogous to solid papillary carcinoma of breast. AB - We describe two cases of a distinctive in situ and invasive cutaneous adnexal neoplasm occurring in the eyelid. Mucinous carcinoma represented the invasive portion of the tumor in one case, whereas the other infiltrated in small solid nests. The in situ component is identical to the recently described solid papillary carcinoma of the breast (endocrine ductal carcinoma in situ). Both tumors produced intra- and extracellular mucin, exhibited endocrine differentiation by immunohistochemistry and ultrastructural analysis, and were positive for estrogen and progesterone receptors. PMID- 9414196 TI - Stage IA uterine serous carcinoma: a study of 13 cases. AB - Although in endometrioid type endometrial carcinoma depth of invasion is a powerful predictor of extrauterine disease and survival, in serous carcinoma its importance is unclear. Recurrences and death in patients with serous tumors confined to the endometrium or an endometrial polyp have been reported. In other studies, however, the absence of myometrial invasion was correlated with a more favorable course. In an attempt to clarify this issue, we reviewed 13 completely staged, stage IA serous carcinomas with follow-up from 10 to 93 months (median 38), in which extensive histologic examination had been performed. Serous carcinoma was identified in an endometrial polyp in six cases, in an endometrial polyp and associated endometrium in four, and solely in the endometrium in three cases. No other histologic types of endometrial carcinoma were present, and there was no myometrial invasion. Multifocal serous intraepithelial carcinoma was also seen in 12 cases. Two of the patients died of disease with intraabdominal carcinomatosis at 10 and 14 months after presentation. The overall estimated survival was 83%, showing a relatively favorable prognosis. In conclusion, although the absence of histologically detected myometrial invasion may be associated with recurrences and death in serous carcinoma, an accurately assessed stage based on a careful histologic examination appears to be, at present, the most reliable predictor of survival. PMID- 9414197 TI - Malignant peripheral nerve sheath tumor of the pleura with epithelial and rhabdomyoblastic differentiation: report of a case clinically simulating mesothelioma. AB - A primary malignant peripheral nerve sheath tumor (MPNST) of the pleura that clinically mimicked a malignant mesothelioma in a 57-year-old man is described. Histologically, the tumor had features similar to those described in cases of the so-called epithelioid MPNST. A unique finding in this case was the demonstration of keratin expression in the epithelioid component of the tumor, as well as the presence of rhabdomyoblasts. This is the first example of an MPNST with heterologous elements arising in the pleura. Immunohistochemical and ultrastructural studies were important in differentiating this tumor from other malignancies with sarcomatoid and epithelioid features involving the pleura. PMID- 9414198 TI - Sharon Whelan Weiss, M.D., A.B., recipient of the 1997 Fred W. Stewart Award. PMID- 9414199 TI - Langerhans cell granulomatosis. PMID- 9414200 TI - Minimal residual cancer. PMID- 9414201 TI - Ice-binding mechanism of winter flounder antifreeze proteins. AB - We have studied the winter flounder antifreeze protein (AFP) and two of its mutants using molecular dynamics simulation techniques. The simulations were performed under four conditions: in the gas phase, solvated by water, adsorbed on the ice (2021) crystal plane in the gas phase and in aqueous solution. This study provided details of the ice-binding pattern of the winter flounder AFP. Simulation results indicated that the Asp, Asn, and Thr residues in the AFP are important in ice binding and that Asn and Thr as a group bind cooperatively to the ice surface. These ice-binding residues can be collected into four distinct ice-binding regions: Asp-1/Thr-2/Asp-5, Thr-13/Asn-16, Thr-24/Asn-27, and Thr 35/Arg-37. These four regions are 11 residues apart and the repeat distance between them matches the ice lattice constant along the (1102) direction. This match is crucial to ensure that all four groups can interact with the ice surface simultaneously, thereby, enhancing ice binding. These Asx (x = p or n)/Thr regions each form 5-6 hydrogen bonds with the ice surface: Asn forms about three hydrogen bonds with ice molecules located in the step region while Thr forms one to two hydrogen bonds with the ice molecules in the ridge of the (2021) crystal plane. Both the distance between Thr and Asn and the ordering of the two residues are crucial for effective ice binding. The proper sequence is necessary to generate a binding surface that is compatible with the ice surface topology, thus providing a perfect "host/guest" interaction that simultaneously satisfies both hydrogen bonding and van der Waals interactions. The results also show the relation among binding energy, the number of hydrogen bonds, and the activity. The activity is correlated to the binding energy, and in the case of the mutants we have studied the number of hydrogen bonds. The greater the number of the hydrogen bonds the greater the antifreeze activity. The roles van der Waals interactions and the hydrophobic effect play in ice binding are also highlighted. For the latter it is demonstrated that the surface of ice has a clathratelike structure which favors the partitioning of hydrophobic groups to the surface of ice. It is suggested that mutations that involve the deletion of hydrophobic residues (e.g., the Leu residues) will provide insight into the role the hydrophobic effect plays in partitioning these peptides to the surface of ice. PMID- 9414203 TI - Protein dynamics derived from clusters of crystal structures. AB - A method is presented to mathematically extract concerted structural transitions in proteins from collections of crystal structures. The "essential dynamics" procedure is used to filter out small-amplitude fluctuations from such a set of structures; the remaining large conformational changes describe motions such as those important for the uptake/release of substrate/ligand and in catalytic reactions. The method is applied to sets of x-ray structures for a number of proteins, and the results are compared with the results from essential dynamics as applied to molecular dynamics simulations of those proteins. A significant degree of similarity is found, thereby providing a direct experimental basis for the application of such simulations to the description of large concerted motions in proteins. PMID- 9414204 TI - Calcium waves in a model with a random spatially discrete distribution of Ca2+ release sites. AB - We study the propagation of intracellular calcium waves in a model that features Ca2+ release from discrete sites in the endoplasmic reticulum membrane and random spatial distribution of these sites. The results of our simulations qualitatively reproduce the experimentally observed behavior of the waves. When the level of the channel activator inositol trisphosphate is low, the wave undergoes fragmentation and eventually vanishes at a finite distance from the region of initiation, a phenomenon we refer to as an abortive wave. With increasing activator concentration, the mean distance of propagation increases. Above a critical level of activator, the wave becomes stable. We show that the heterogeneous distribution of Ca2+ channels is the cause of this phenomenon. PMID- 9414202 TI - The effects of geometrical parameters on synaptic transmission: a Monte Carlo simulation study. AB - Monte Carlo simulations of transmitter diffusion and its interactions with postsynaptic receptors have been used to study properties of quantal responses at central synapses. Fast synaptic responses characteristic of those recorded at glycinergic junctions on the teleost Mauthner cell (time to peak approximately 0.3-0.4 ms and decay time constant approximately 3-6 ms) served as the initial reference, and smaller contacts with fewer postsynaptic receptors were also modeled. Consistent with experimental findings, diffusion, simulated using a random walk algorithm and assuming a diffusion coefficient of 0.5-1.0 x 10(-5) cm2 s(-1), was sufficiently fast to account for transmitter removal from the synaptic cleft. Transmitter-receptor interactions were modeled as a two-step binding process, with the double-bound state having opened and closed conformations. Addition of a third binding step only slightly decreased response amplitude but significantly slowed both its rising and decay phases. The model allowed us to assess the sources of response variability and the likelihood of postsynaptic saturation as functions of multiple kinetic and spatial parameters. The method of nonstationary fluctuation analysis, typically used to estimate the number of functional channels at a synapse and single channel current, proved unreliable, presumably because the receptors in the postsynaptic matrix are not uniformly exposed to the same profile of transmitter concentration. Thus, the time course of the probability of channel opening most likely varies among receptors. Finally, possible substrates for phenomena of synaptic plasticity, such as long-term potentiation, were explored, including the diameter of the contact zone, defined by the region of pre- and postsynaptic apposition, the number and distribution of the receptors, and the degree of vesicle filling. Surprisingly, response amplitude is quite sensitive to the size of the receptor free annulus surrounding the receptor cluster, such that expansion of the contact zone could produce an appreciable increase in quantal size, normally attributed to either the presence of more receptors or the release of more transmitter molecules. PMID- 9414205 TI - Structure and dynamic properties of diunsaturated 1-palmitoyl-2-linoleoyl-sn glycero-3-phosphatidylcholine lipid bilayer from molecular dynamics simulation. AB - Unsaturated fatty acid chains are known to be an essential structural part of biomembranes, but only monounsaturated chains have been included in the molecular dynamics (MD) simulations of membrane systems. Here we present a 1-ns MD simulation for a diunsaturated 1-palmitoyl-2-linoleoyl-sn-glycero-3 phosphatidylcholine (PLPC; 16:0/18:2[delta9,12]) bilayer. The structural behavior of the phosphatidylcholine headgroup, the glycerol backbone, and the hydrating water were assessed and found to be consistent with the existing information about similar systems from both experimental and computational studies. Further analysis was focused on the structure of the double bond region and the effects of the diunsaturation on the bilayer interior. The behavior of the diunsaturated sn-2 chains is affected by the tilted beginning of the chain and the four main conformations of the double bond region. The double bonds of the sn-2 chains also influenced the characteristics of the saturated chains in the sn-1 position. Furthermore, extreme conformations of the sn-2 chains existed that are likely to be related to the functional role of the double bonds. The results here point out the importance of polyunsaturation for the biological interpretations deduced from the membrane MD simulations. PMID- 9414207 TI - Residence time calculation for chemotactic bacteria within porous media. AB - Local chemical gradients can have a significant impact on bacterial population distributions within subsurface environments by evoking chemotactic responses. These local gradients may be created by consumption of a slowly diffusing nutrient, generation of a local food source from cell lysis, or dissolution of nonaqueous phase liquids trapped within the interstices of a soil matrix. We used a random walk simulation algorithm to study the effect of a local microscopic gradient on the swimming behavior of bacteria in a porous medium. The model porous medium was constructed using molecular dynamics simulations applied to a fluid of equal-sized spheres. The chemoattractant gradient was approximated with spherical symmetry, and the parameters for the swimming behavior of soil bacterium Pseudomonas putida were based on literature values. Two different mechanisms for bacterial chemotaxis, one in which the bacteria responded to both positive and negative gradients, and the other in which they responded only to positive gradients, were compared. The results of the computer simulations showed that chemotaxis can increase migration through a porous medium in response to microscopic-scale gradients. The simulation results also suggested that a more significant role of chemotaxis may be to increase the residence time of the bacteria in the vicinity of an attractant source. PMID- 9414206 TI - Highly aligned lipid membrane systems in the physiologically relevant "excess water" condition. AB - The "excess water" condition in biologically relevant systems is met when a membrane mesophase coexists with excess bulk water. Further addition of water to such a system results in no change to any of the system's physical properties (e.g., transition temperature, repeat spacing, and structural mesophases). Moreover, because biological membranes are anisotropic systems, many of their properties are best studied using aligned samples. Although model membrane systems are routinely aligned, they have traditionally been hydrated with water vapor. It is well known that membranes exposed to water vapor at 100% humidity do not imbibe the same quantity of water as a sample in contact with liquid water. As such, membranes that have been hydrated with water vapor have physical properties different from those of membranes dispersed in water. Because of this shortcoming, aligned membranes have not been utilized to their full potential. Here we present a novel and simple method of aligning model membrane systems under conditions of excess water, which will make possible, for the first time, a variety of techniques (e.g., neutron and x-ray diffraction, nuclear magnetic resonance, electron spin resonance, attenuated total reflection infrared spectroscopy, etc.) for studying such systems under physiologically relevant conditions. In addition, when dealing with samples of limited availability, the system allows for the conditions (buffer pH and ionic strength) to be altered without any effect on the sample's alignment. PMID- 9414208 TI - A soft-modeling approach to interpret thermodynamic and conformational transitions of polynucleotides. AB - Multivariate outputs from the experimental monitoring of biochemical processes are usually difficult to interpret applying methods based on a priori chemical models. Curve resolution methods are model-free procedures, generally known as soft-modeling methods, which obtain the concentration profiles and instrumental responses of each individual species involved in a multivariate monitored process without making any kind of external assumption. Of the curve resolution methods available, the alternating least squares (ALS) is proposed here because of its ability to operate on one or on several matrices. Furthermore, ALS allows the introduction of information related to the internal data structure and to the general features of the concentration profiles and instrumental responses through the input of suitable constraints in the iterative resolution procedure. The ALS potential is tested on several data sets coming from the multivariate spectrometric monitoring of polyuridylic (polyU), polycytidylic (polyC), and polyadenylic (polyA) protonation equilibria in dioxane/water 30% (v/v). Information concerning the evolution of the concentration profiles and the spectra of each individual species involved in the acid-base equilibria, the presence and pattern of polyelectrolyte effects, and the presence of conformational transitions associated or not with the proton uptake process is presented. PMID- 9414209 TI - Calculation of diffusion-limited kinetics for the reactions in collision coupling and receptor cross-linking. AB - Both enzyme (e.g., G-protein) activation via a collision coupling model and the formation of cross-linked receptors by a multivalent ligand involve reactions between two molecules diffusing in the plasma membrane. The diffusion of these molecules is thought to play a critical role in these two early signal transduction events. In reduced dimensions, however, diffusion is not an effective mixing mechanism; consequently, zones in which the concentration of particular molecules (e.g., enzymes, receptors) becomes depleted or enriched may form. To examine the formation of these depletion/ accumulation zones and their effect on reaction rates and ultimately the cellular response, Monte Carlo techniques are used to simulate the reaction and diffusion of molecules in the plasma membrane. The effective reaction rate at steady state is determined in terms of the physical properties of the tissue and ligand for both enzyme activation via collision coupling and the generation of cross-linked receptors. The diffusion-limited reaction rate constant is shown to scale with the mean square displacement of a receptor-ligand complex. The rate constants determined in the simulation are compared with other theoretical predictions as well as experimental data. PMID- 9414210 TI - Temperature, stability, and the hydrophobic interaction. AB - Changes in free energy are normally used to track the effect of temperature on the stability of proteins and hydrophobic interactions. Use of this procedure on the aqueous solubility of hydrocarbons, a standard representation of the hydrophobic effect, leads to the conclusion that the hydrophobic effect increases in strength as the temperature is raised to approximately 140 degrees C. Acceptance of this interpretation leads to a number of far-reaching conclusions that are at variance with the original conception of the hydrophobic effect and add considerably to the complexity of interpretation. There are two legitimate thermodynamic functions that can be used to look at stability as a function of temperature: the standard Gibbs free energy change, deltaG degrees, and deltaG degrees/T. The latter is proportional to the log of the equilibrium constant and is sometimes called the Massieu-Planck function. Arguments are presented for using deltaG degrees/T rather than deltaG degrees for variations in stability with temperature. This makes a considerable difference in the interpretation of the hydrophobic interaction, but makes little change in the stability profile of proteins. Protein unfolding and the aqueous solubility of benzene are given as examples. The contrast between protein unfolding and the hydration of nonpolar molecules provides a rough estimate of the contribution of other factors that stabilize and destabilize protein structure. PMID- 9414211 TI - Current noise spectrum and capacitance due to the membrane motor of the outer hair cell: theory. AB - The voltage-dependent motility of the outer hair cell is based on a membrane motor densely distributed in the lateral membrane. The gating charge of the membrane motor is manifested as a bell-shaped membrane potential dependence of the membrane capacitance. In this paper it is shown that movements of the gating charge should produce a high-pass current noise described by an inverse Lorentzian similar to the one shown by Kolb and Lauger for ion carriers. The frequency dependence of the voltage-dependent capacitance is also derived. These derivations are based on membrane motor models with two or three states. These two models lead to similar predictions on the capacitance and current noise. It is expected that the examination of the spectral properties of these quantities would be a useful means of determining the relaxation time for conformational transitions of the membrane motor. PMID- 9414212 TI - Binding pathway of retinal to bacterio-opsin: a prediction by molecular dynamics simulations. AB - Formation of bacteriorhodopsin (bR) from apoprotein and retinal has been studied experimentally, but the actual pathway, including the point of entry, is little understood. Molecular dynamics simulations provide a surprisingly clear prediction. A window between bR helices E and F in the transmembrane part of the protein can be identified as an entry point for retinal. Steered molecular dynamics, performed by applying a series of external forces in the range of 200 1000 pN over a period of 0.2 ns to retinal, allows one to extract this chromophore from bR once the Schiff base bond to Lys216 is cleaved. Extraction proceeds until the retinal tail forms a hydrogen bond network with Ala144, Met145, and Ser183 side groups lining the exit/entry window. The manipulation induces a distortion with a fitted root mean square deviation of coordinates (ignoring retinal, water, and hydrogen atoms) of less than 1.9 A by the time the retinal carbonyl reaches the protein surface. The forces needed to extract retinal are due to friction and do not indicate significant potential barriers. The simulations therefore suggest a pathway for the binding of retinal. Water molecules are found to play a crucial role in the binding process. PMID- 9414213 TI - Influence of the membrane potential on the free energy of an intrinsic protein. AB - A modified Poisson-Boltzmann equation is developed from statistical mechanical considerations to describe the influence of the transmembrane potential on macromolecular systems. Using a Green's function formalism, the electrostatic free energy of a protein associated with the membrane is expressed as the sum of three terms: a contribution from the energy required to charge the system's capacitance, a contribution corresponding to the interaction of the protein charges with the membrane potential, and a contribution corresponding to a voltage-independent reaction field free energy. The membrane potential, which is due to the polarization interface, is calculated in the absence of the protein charges, whereas the reaction field is calculated in the absence of transmembrane potential. Variations in the capacitive energy associated with typical molecular processes are negligible under physiological conditions. The formulation of the theory is closely related to standard algorithms used to solve the Poisson Boltzmann equation and only small modifications to current source codes are required for its implementation. The theory is illustrated by examining the voltage-dependent membrane insertion of a simple polyalanine alpha-helix and by computing the electrostatic potential across a 60-A-diameter sphere meant to represent a large intrinsic protein. PMID- 9414214 TI - Analysis of the stability of looped-out and stacked-in conformations of an adenine bulge in DNA using a continuum model for solvent and ions. AB - A combination of conformational search, energy minimization, and energetic evaluation using a continuum solvent treatment has been employed to study the stability of various conformations of the DNA fragment d(CGCAGAA)/d(TTCGCG) containing a single adenine bulge. The extra-helical (looped-out) bulge conformation derived from a published x-ray structure and intra-helical (stacked bulge base) model structures partially based on nuclear magnetic resonance (NMR) data were used as start structures for the conformational search. Solvent dependent contributions to the stability of the conformations were calculated from the solvent exposed molecular surface area and by using the finite difference Poisson-Boltzmann approach. Three classes (I-III) of bulge conformations with calculated low energies can be distinguished. The lowest energy conformations were found in class I, corresponding to structures with the bulge base stacked between flanking helices, and class II, composed of structures forming a triplet of the bulge base and a flanking base pair. All extra-helical bulge structures, forming class III, were found to be less stable compared with the lowest energy structures of class I and II. The results are consistent with NMR data on an adenine bulge in the same sequence context indicating an intra helical or triplet bulge conformation in solution. Although the total energies and total electrostatic energies of the low-energy conformations show only relatively modest variations, the energetic contributions to the stability were found to vary significantly among the classes of bulge structures. All intra helical bulge structures are stabilized by a more favorable Coulomb charge-charge interaction but destabilized by a larger electrostatic reaction field contribution compared with all extra-helical and most triplet bulge structures. Van der Waals packing interactions and nonpolar surface-area-dependent contributions appear to favor triplet class II structures and to a lesser degree also the intra-helical stacked bulge conformations. The large conformational variation found for class III conformers might add a favorable entropic contribution to the stability of the extra-helical bulge form. PMID- 9414215 TI - Electrotonic measurements by electric field-induced polarization in neurons: theory and experimental estimation. AB - We present a theory for estimation of the dendritic electrotonic length constant and the membrane time constant from the transmembrane potential (TMP) induced by an applied electric field. The theory is adapted to morphologically defined neurons with homogeneous passive electric properties. Frequency characteristics and transients at the onset and offset of the DC field are considered. Two relations are useful for estimating the electrotonic parameters: 1) steady-state polarization versus the dendritic electrotonic length constant; 2) membrane time constant versus length constant. These relations are monotonic and may provide a unique estimate of the electrotonic parameters for 3D-reconstructed neurons. Equivalent tip-to-tip electrotonic length of the dendritic tree was estimated by measuring the equalization time of the field-induced TMP. For 11 turtle spinal motoneurons, the electrotonic length from tip to tip of the dendrites was in the range of 1-2.5 lambda, whereas classical estimation using injection of current pulses gave an average dendrite length of 0.9-1.1 lambda. For seven ventral horn interneurons, the estimates were 0.7-2.6 lambda and 0.6-0.9 lambda, respectively. The measurements of the field-induced polarization promise to be a useful addition to the conventional methods using microelectrode stimulation. PMID- 9414216 TI - Cryo-transmission electron microscopy of a superstructure of fluid dioleoylphosphatidylcholine (DOPC) membranes. AB - Using cryo-transmission electron microscopy, we have obtained abundant and reproducible evidence for a superstructure of dioleoylphosphatidylcholine (DOPC) bilayers. Dispersions of vesicles were prepared by gentle shaking of a 2% suspension of DOPC in water followed in part by extrusion through a porous technical membrane. Sampling and cryofixation took place at various times within 3 weeks after the preparation. From the micrographs we infer that the small fraction of vesicles enclosing one another develop passages (connections) between the bilayers. In contrast, the superstructure is basically a feature of disconnected membranes. Among its modifications are isolated membrane bends or folds and a grainy membrane texture with a minimal grain spacing of 4-6 nm. In the extruded dispersions the passages and the superstructure seem to be formed mostly within the first day. The fraction of smooth and unilamellar vesicles is large at all times and in all dispersions. PMID- 9414218 TI - Structure and dynamics of an amphiphilic peptide in a lipid bilayer: a molecular dynamics study. AB - A molecular dynamics simulation of a simple model membrane system composed of a single amphiphilic helical peptide (ace-K2GL16K2A-amide) in a fully hydrated 1,2 dimyristoyl-sn-glycero-3-phosphocholine bilayer was performed for a total of 1060 ps. The secondary structure of the peptide and its stability were described in terms of average dihedral angles, phi and psi, and the C alpha torsion angles formed by backbone atoms; by the average translation per residue along the helix axis; and by the intramolecular peptide hydrogen bonds. The results indicated that residues 6 through 15 remain in a stable right-handed alpha-helical conformation, whereas both termini exhibit substantial fluctuations. A change in the backbone dihedral angles for residues 16 and 17 is accompanied by the loss of two intramolecular hydrogen bonds, leading to a local but long-lived disruption of the helix. The dynamics of the peptide was characterized in terms of local and global helix motions. The local motions of the N-H bond angles were described in terms of the autocorrelation functions of P2[cos thetaNH(t, t + tau)] and reflected the different degrees of local peptide order as well as a variation in time scale for local motions. The chi1 and chi2 dihedral angles of the leucine side chains underwent frequent transitions between potential minima. No connection between the side-chain positions and their mobility was observed, however. In contrast, the lysine side chains displayed little mobility during the simulation. The global peptide motions were characterized by the tilting and bending motions of the helix. Although the peptide was initially aligned parallel to the bilayer normal, during the simulation it was observed to tilt away from the normal, reaching an angle of approximately 25 degrees by the end of the simulation. In addition, a slight bend of the helix was detected. Finally, the solvation of the peptide backbone and side-chain atoms was also investigated. PMID- 9414217 TI - Trehalose-induced destabilization of interdigitated gel phase in dihexadecylphosphatidylcholine. AB - Trehalose is believed to have the ability to protect some organisms against low temperatures. To clarify the cryoprotective mechanism of trehalose, the structure and the phase behavior of fully hydrated dihexadecylphosphatidylcholine (DHPC) membranes in the presence of various concentrations of trehalose were studied by means of differential scanning calorimetry (DSC), static x-ray diffraction, and simultaneous x-ray diffraction and DSC measurements. The temperature of the interdigitated gel (Lbeta(i))-to-ripple (Pbeta') phase transition of DHPC decreases with a rise in trehalose concentration up to approximately 1.0 M. Above a trehalose concentration of approximately 1.0 M, no Lbeta(i) phase is observed. In this connection, the electron density profile calculated from the lamellar diffraction data in the presence of 1.6 M trehalose indicates that DHPC forms noninterdigitated bilayers below the P beta' phase. It was concluded that trehalose destabilizes the Lbeta(i) phase of DHPC bilayers. This suggests that trehalose reduces the area at the interface between the lipid and water. The relation between this effect of trehalose and a low temperature tolerance was discussed from the viewpoint of cold-induced denaturation of proteins. PMID- 9414219 TI - Amplification of electromagnetic signals by ion channels. AB - Cells may respond to the exposure of low-frequency electromagnetic fields with changes in cell division, ion influx, chemical reaction rates, etc. The chain of events leading to such responses is difficult to study, mainly because of extremely small energies associated with low-frequency fields, usually much smaller than the thermal noise level. However, the presence of stochastic systems (for instance, ion channels) provides a basis for signal amplification, and could therefore, despite the low signal-to-noise ratio of the primary response, lead to the transmission of weak signals along the signaling pathways of cells. We have explored this possibility for an ion channel model, and we present a theory, based on the formalism of stochastically driven processes, that relates the time averages of the ion channel currents to the amplitude and frequency of the applied signal. It is concluded from this theory that the signal-to-noise ratio increases with the number of channels, the magnitude of the rate constants, and the frequency response of the intracellular sensing system (for instance, a calcium oscillator). The amplification properties of the stochastic system are further deduced from numerical simulations carried out on the model, which consists of multiple identical two-state channels, and the behavior for different parameters is examined. Numerical estimates of the parameters show that under optimum conditions, even very weak low-frequency electromagnetic signals (<100 Hz and down to 100 microT) may be detected in a cellular system with a large number of ion channels. PMID- 9414220 TI - Characterization of biomimetic surfaces formed from cell membranes. AB - A method for fabricating biomimetic surfaces from intact cell membranes is described. A monolayer of alkanethiol on gold is covered by a second layer derived from the components of erythrocyte membranes either by self-assembly or by Langmuir-Blodgett methods. The resulting asymmetric hybrid layer was characterized by ellipsometry, surface plasmon resonance (SPR), contact angle, capacitance, voltammetry, and electron and atomic force microscopy. The erythrocyte membrane layer was measured to be approximately 30-40 A in thickness. Using SPR, the presence of erythrocyte components on the surface was demonstrated by their selective removal by enzymatic action. The uniform deposition of membranous material on the substrate was shown by electron and atomic force microscopy. Demonstration of acetylcholinesterase (AChase) activity, a membrane anchored enzyme, on the surface for at least 8 days, suggests that the outer leaflet of the erythrocyte membrane is present in its native form. Cyclic voltammetry demonstrates that enhanced electron transport from a solution redox species accompanies formation of the erythrocyte layer at the surface. This enhanced electron transport is blocked by 4,4'-diisothiocyanate stilbene-2,2' disulfonic acid, a well known blocker of anion transport, suggesting that an erythrocyte anion transporter protein is incorporated into the surface layer in an active conformation. PMID- 9414221 TI - Influence of membrane-spanning alpha-helical peptides on the phase behavior of the dioleoylphosphatidylcholine/water system. AB - The effect of solubilized hydrophobic peptides on the phase behavior of dioleoylphosphatidylcholine (DOPC)/water system was studied by 2H- and 31P-NMR spectroscopy and by x-ray diffraction, and partial phase diagrams were constructed. The utilized peptides were HCO-AWW(LA)5WWA-NHCH2CH2OH (WALP16), which is an artificial peptide designed to resemble a transmembrane part of a membrane protein; and VEYAGIALFFVAAVLTLWSMLQYLSAAR (Pgs peptide E), a peptide that is identical to one of the putative transmembrane segments of the membrane associated protein phosphatidylglycerophosphate synthase (Pgs) in Escherichia coli. Circular dichroism spectroscopy suggests that both peptides are mostly alpha-helical in DOPC vesicles. The most striking features in the phase diagram of the WALP16/DOPC/water system are 1) a single lamellar liquid crystalline (L alpha) phase forms only at very low peptide concentrations. 2) At low water content and above a peptide/lipid molar ratio of approximately 1:75 a reversed hexagonal liquid crystalline (H[II]) phase coexists with an L alpha phase, while in excess water this phase forms at a peptide/lipid molar ratio of approximately 1:25. 3) At peptide/lipid ratios > or =1:6 a single H(II) phase is stable. Also, the Pgs peptide E strongly affects the phase behavior, and a single L alpha phase is only found at low peptide concentrations (peptide/lipid molar ratios <1:50), and water concentrations <45% (w/w). Higher peptide content results in coexistence of L alpha and isotropic phases. Generally, the fraction of the isotropic phase increases with increasing temperature and water concentration, and at 80% (w/w) water content only a single isotropic phase is stable at 55 degrees C. Thus, both peptides were found to be able to induce nonlamellar phases, although different in structure, in the DOPC/water system. The phase transitions, the extensions of the one-phase regions, and the phase structures observed for the two systems are discussed in terms of the molecular structure of the two peptides and the matching between the hydrophobic lengths of the peptides and the bilayer thickness of DOPC. PMID- 9414222 TI - The mechanism of lamellar-to-inverted hexagonal phase transitions in phosphatidylethanolamine: implications for membrane fusion mechanisms. AB - We studied the mechanism of the lamellar-to-inverted hexagonal (L alpha/H[II]) phase transition, using time-resolved cryotransmission electron microscopy (TRC TEM), 31P-NMR, and differential scanning calorimetry. The transition was initiated in dispersions of large unilamellar vesicles of dipalmitoleoyl phosphatidylethanolamine (DiPoPE). We present evidence that the transition proceeds in three steps. First, many small connections form between apposed membranes. Second, the connections aggregate within the planes of the bilayers, forming arrays with hexagonal order in some projections. Third, these quasihexagonal structures elongate into small domains of H(II) phase, acquiring lipid molecules by diffusion from contiguous bilayers. A previously proposed membrane fusion mechanism rationalizes these results. The modified stalk theory predicts that the L alpha/H(II) phase transition involves some of the same intermediate structures as membrane fusion. The small interbilayer connections observed via TRC-TEM are compatible with the structure of a critical intermediate in the modified stalk mechanism: the trans monolayer contact (TMC). The theory predicts that 1) TMCs should form starting at tens of degrees below TH; 2) when TMCs become sufficiently numerous, they should aggregate into transient arrays like the quasihexagonal arrays observed here by TRC-TEM; and 3) these quasihexagonal arrays can then elongate directly into H(II) phase domains. These predictions rationalize the principal features of our data, which are incompatible with the other transition mechanisms proposed to date. Thus these results support the modified stalk mechanism for both membrane fusion and the L alpha/H(II) phase transition. We also discuss some implications of the modified stalk theory for fusion in protein-containing systems. Specifically, we point out that recent data on the effects of hydrophobic peptides and viral fusion peptides on lipid phase behavior are consistent with an effect of the peptides on TMC stability. PMID- 9414223 TI - Partitioning of a fluorescent phospholipid between fluid bilayers: dependence on host lipid acyl chains. AB - The partition coefficient Kp was measured for a headgroup-labeled phospholipid (12:0,12:0)-N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-PE (12-NBD-PE), equilibrated between LUV of a series of phosphatidylcholines (PC). Fluorescence resonance energy transfer between the 12-NBD-PE and a headgroup-rhodamine-labeled PE was used to find the equilibrium concentration of the 12-NBD-PE in the different LUV. Reliable equilibrium concentrations were obtained by monitoring the approach to equilibrium starting from a concentration below and from a concentration above the ultimate values. Using (16:0,18:1delta9)-PC as the reference lipid, Kp ranged from a high value of 1.65 favoring (16:0,18:1delta9)-PC over (16:1delta9,16:1delta9)-PC, to a low value of 0.90, favoring (22:1delta13,22:1delta13)-PC over (16:0,18:1delta9)-PC. The Kp values enabled calculation of the acyl chain contribution to the excess free energy of mixing for (12:0,12:0) acyl chains at infinite dilution in the L alpha phase of PC having acyl chains of (16:0,18:1delta9), (16:1delta9,16:1delta9), (18:1delta9,18:1delta9), (18:1delta6,18:1delta6), (20:1delta11,20:1delta11), and (22:1delta13,22:1delta13). (14:1delta9,14:1delta9)-PC was found to transfer so rapidly between LUV as to preclude reliable Kp measurement. PMID- 9414224 TI - Phosphorylation-dependent power output of transgenic flies: an integrated study. AB - We examine how the structure and function of indirect flight muscle (IFM) and the entire flight system of Drosophila melanogaster are affected by phosphorylation of the myosin regulatory light chain (MLC2). This integrated study uses site directed mutagenesis to examine the relationship between removal of the myosin light chain kinase (MLCK) phosphorylation site, in vivo function of the flight system (flight tests, wing kinematics, metabolism, power output), isolated IFM fiber mechanics, MLC2 isoform pattern, and sarcomeric ultrastructure. The MLC2 mutants exhibit graded impairment of flight ability that correlates with a reduction in both IFM and flight system power output and a reduction in the constitutive level of MLC2 phosphorylation. The MLC2 mutants have wild-type IFM sarcomere and cross-bridge structures, ruling out obvious changes in the ultrastructure as the cause of the reduced performance. We describe a viscoelastic model of cross-bridge dynamics based on sinusoidal length perturbation analysis (Nyquist plots) of skinned IFM fibers. The sinusoidal analysis suggests the high power output of Drosophila IFM required for flight results from a phosphorylation-dependent recruitment of power-generating cross bridges rather than a change in kinetics of the power generating step. The reduction in cross-bridge number appears to affect the way mutant flies generate flight forces of sufficient magnitude to keep them airborne. In two MLC2 mutant strains that exhibit a reduced IFM power output, flies appear to compensate by lowering wingbeat frequency and by elevating wingstroke amplitude (and presumably muscle strain). This behavioral alteration is not seen in another mutant strain in which the power output and estimated number of recruited cross-bridges is similar to that of wild type. PMID- 9414225 TI - Sequence effects on the relative thermodynamic stabilities of B-Z junction forming DNA oligomeric duplexes. AB - Circular dichroism (CD) and ultraviolet absorption techniques were employed in characterizing the sequence-dependent thermodynamic stabilities of B-Z junction forming DNA duplexes. The Watson strand of the duplexes has the general sequence (5meC-G)4-NXYG-ACTG (where N = A or G and XY represents all permutations of pyrimidine bases). Duplexes were generated by mixing stoichiometric amounts of the complementary strands. Circular dichroism studies indicate that each duplex is fully right-handed at low salt (e.g., 115 mM Na+) but undergoes a salt-induced conformational transition to a structure that possesses both left- and right handed conformations at high salt (4.5 M Na+), and hence a B-Z junction. Optical melting studies of the DNA duplexes at fixed DNA concentration with total Na+ concentration ranging from 15 mM to 5.0 M were determined. A nonlinear dependence of the melting temperature (Tm) on [Na+] was observed. Thermodynamic parameters at Na+ concentrations of 115 mM and 4.5 M with a wide range of DNA concentrations were determined from UV optical melting studies via construction of van't Hoff plots. A change of a single dinucleotide within these duplexes significantly affected the helix stabilities. The experimentally obtained free energies for the duplex to single-strand transitions were in close agreement with predicted values obtained from two different methods. PMID- 9414226 TI - Simulation of the conformation and dynamics of a double-helical model for DNA. AB - We propose a partially flexible, double-helical model for describing the conformational and dynamic properties of DNA. In this model, each nucleotide is represented by one element (bead), and the known geometrical features of the double helix are incorporated in the equilibrium conformation. Each bead is connected to a few neighbor beads in both strands by means of stiff springs that maintain the connectivity but still allow for some extent of flexibility and internal motion. We have used Brownian dynamics simulation to sample the conformational space and monitor the overall and internal dynamics of short DNA pieces, with up to 20 basepairs. From Brownian trajectories, we calculate the dimensions of the helix and estimate its persistence length. We obtain translational diffusion coefficient and various rotational relaxation times, including both overall rotation and internal motion. Although we have not carried out a detailed parameterization of the model, the calculated properties agree rather well with experimental data available for those oligomers. PMID- 9414227 TI - DNA length and concentration dependencies of anisotropic phase transitions of DNA solutions. AB - Critical concentrations for the isotropic to cholesteric phase transitions of double-stranded DNA fragments in simple buffered saline (0.1 M NaCl) solutions were determined as a function of DNA contour length ranging from approximately 50 nm to 2700 nm, by solid-state 31P NMR spectroscopy and polarized light microscopy. As expected for semirigid chains, the critical concentrations decrease sharply with increasing DNA length near the persistence length in the range from 50 to 110 nm, and approach a plateau when the contour length exceeds 190 nm. The biphasic region is substantially wider than observed for xanthan, another semirigid polyelectrolyte approximately twice as stiff as DNA, primarily because of low critical concentrations for first appearance of the anisotropic phase, C(i)*, in DNA samples > or =110 nm (320 base pairs) long. The limiting C(i)* for DNA > or =490 nm long is exceptionally low (only 13 mg/ml) and is substantially lower than the C(i)* of approximately 40 mg/ml reported for the stiffer xanthan polyelectrolyte. The much higher values of the critical concentrations, C(a)*, for the disappearance of the isotropic DNA phase (> or =67 mg/ml) are modestly higher than those observed for xanthan and are predicted reasonably well by a theory that has been applied to other semirigid polymers, if a DNA persistence length in the consensus range of 50-100 nm is assumed. By contrast, the broad biphasic region and low C(i)* values of DNA fragments > or =190 nm long could only be reconciled with theory by assuming persistence lengths of 200-400 nm. The latter discrepancies are presumed to reflect some combination of deficiencies in current theory as applied to chiral, strong polyelectrolytes such as DNA, and sequence-dependent variations in DNA properties such as flexibility, curvature, or interaction potential. The propensity of DNA to spontaneously self-order at low concentrations well in the physiological range may have biological significance. PMID- 9414228 TI - Optical rotation of the second harmonic radiation from retinal in bacteriorhodopsin monomers in Langmuir-Blodgett film: evidence for nonplanar retinal structure. AB - We observed optical rotation of the plane of polarization of the second harmonic (SH) radiation at 532 nm (in resonance with the retinal absorption) generated in reflection geometry in Langmuir-Blodgett film of bacteriorhodopsin (bR). The analysis of the experimental data showed that this effect arises from the nonvanishing contribution of the antisymmetrical part of the hyperpolarizability tensor. This requires that the dipole moment of the resonant electronic transition, the change of the dipole moment upon electronic excitation, and the long axis of the retinal not be coplanar. Such conditions are satisfied only if the retinal has a nonplanar geometry, a conclusion that could lend support to the heterogeneity model of the origin of the biphasic band shape of the linear CD spectrum of the retinal in bR. On the basis of our theoretical analysis, we were able to estimate the angle between the induced dipole moment and the plan that contains the long axis of the chromophore and the transition dipole moment of the retinal absorption. PMID- 9414229 TI - Evidence for the first phase of the reprotonation switch of bacteriorhodopsin from time-resolved photovoltage and flash photolysis experiments on the photoreversal of the M-intermediate. AB - The kinetics of the photoreversal reaction of the M-intermediate of bacteriorhodopsin (bR) was investigated by time-resolved optical absorption spectroscopy and photovoltage measurements using double-flash excitation (a green flash (532 nm) followed by a blue flash (400 nm) after a variable delay). The sign of the photovoltage and the 1H/2H kinetic isotope effect indicate that the Schiff base is reprotonated by a group between the Schiff base and the extracellular surface, probably Asp85. Analysis of the kinetic data shows that the charge movement in 150 mM KCl at 12 degrees C is characterized by two components with time constants of approximately 100 ns and approximately 600 ns, respectively, which are independent of the delay time between the flashes and the pH. The amplitudes of the fast and slow components depend on the delay and the pH. The slower component starts to contribute to the charge movement only after delays longer than 100 micros, is absent at low pH, and increases in amplitude with a pKa of approximately 6. Because the proton release group deprotonates after 70-100 micros and has a transient pKa of 5.8, these results suggest the following assignment: the fast and the combination of fast and slow components represent photoreversal from two M states, with the release group protonated and deprotonated, respectively. The slow phase of the photoreversal starts from a state with the release group deprotonated, and with the pK of Asp85 elevated, and is probably due to the restoration of the pK of Asp85 to its initial low value. This provides further evidence for coupling between the pK's of Asp85 and the release group and suggests that proton release is the first step in the reprotonation switch. At alkaline pH the amplitude of the electrical signal from the back photoreaction decreases with an apparent pK of 8, without a corresponding decrease in the amount of M. At neutral pH the movement of the positively charged guanidinium group of Arg82 from a position near the release group on the surface to Asp85 makes a substantial contribution to the electrical photoreversal amplitude. Above the pK of the release group in the unphotolysed state (approximately 8), Arg82 stays near the surface, leading to a corresponding signal reduction. PMID- 9414231 TI - Statistical mechanics of simple models of protein folding and design. AB - It is now believed that the primary equilibrium aspects of simple models of protein folding are understood theoretically. However, current theories often resort to rather heavy mathematics to overcome some technical difficulties inherent in the problem or start from a phenomenological model. To this end, we take a new approach in this pedagogical review of the statistical mechanics of protein folding. The benefit of our approach is a drastic mathematical simplification of the theory, without resort to any new approximations or phenomenological prescriptions. Indeed, the results we obtain agree precisely with previous calculations. Because of this simplification, we are able to present here a thorough and self contained treatment of the problem. Topics discussed include the statistical mechanics of the random energy model (REM), tests of the validity of REM as a model for heteropolymer freezing, freezing transition of random sequences, phase diagram of designed ("minimally frustrated") sequences, and the degree to which errors in the interactions employed in simulations of either folding and design can still lead to correct folding behavior. PMID- 9414230 TI - A comparison of the efficiency of G protein activation by ligand-free and light activated forms of rhodopsin. AB - Activation of the photoreceptor G protein transducin (Gt) by opsin, the ligand free form of rhodopsin, was measured using rod outer segment membranes with densities of opsin and Gt similar to those found in rod cells. When GTPgammaS was used as the activating nucleotide, opsin catalyzed transducin activation with an exponential time course with a rate constant k(act) on the order of 2 x 10(-3)s( 1). Comparison under these conditions to activation by flash-generated metarhodopsin II (MII) revealed that opsin- and R*-catalyzed activation showed similar kinetics when MII was present at a surface density approximately 10(-6) lower than that of opsin. Thus, in contrast to some previous reports, we find that the catalytic potency of opsin is only approximately 10(-6) that of MII. In the presence of residual retinaldehyde-derived species present in membranes treated with hydroxylamine after bleaching, the apparent k(act) observed was much higher than that for opsin, suggesting a possible explanation for previous reports of more efficient activation by opsin. These results are important for considering the possible role of opsin in the diverse phenomena in which it has been suggested to play a key role, such as bleaching desensitization and retinal degeneration induced by continuous light or vitamin A deprivation. PMID- 9414232 TI - Interactions of lysozyme in concentrated electrolyte solutions from dynamic light scattering measurements. AB - The diffusion of hen egg-white lysozyme has been studied by dynamic light scattering in aqueous solutions of ammonium sulfate as a function of protein concentration to 30 g/liter. Experiments were conducted under the following conditions: pH 4-7 and ionic strength 0.05-5.0 M. Diffusivity data for ionic strengths up to 0.5 M were interpreted in the context of a two-body interaction model for monomers. From this analysis, two potential-of-mean-force parameters, the effective monomer charge, and the Hamaker constant were obtained. At higher ionic strength, the data were analyzed using a model that describes the diffusion coefficient of a polydisperse system of interacting protein aggregates in terms of an isodesmic, indefinite aggregation equilibrium constant. Data analysis incorporated multicomponent virial and hydrodynamic effects. The resulting equilibrium constants indicate that lysozyme does not aggregate significantly as ionic strength increases, even at salt concentrations near the point of salting out precipitation. PMID- 9414233 TI - Temperature dependence of Q-band electron paramagnetic resonance spectra of nitrosyl heme proteins. AB - The Q-band (35 GHz) electron paramagnetic resonance (EPR) spectra of nitrosyl hemoglobin (HbNO) and nitrosyl myoglobin (MbNO) were studied as a function of temperature between 19 K and 200 K. The spectra of both heme proteins show two classes of variations as a function of temperature. The first one has previously been associated with the existence of two paramagnetic species, one with rhombic and the other with axial symmetry. The second one manifests itself in changes in the g-factors and linewidths of each species. These changes are correlated with the conformational substates model and associate the variations of g-values with changes in the angle of the N(his)-Fe-N(NO) bond in the rhombic species and with changes in the distance between Fe and N of the proximal (F8) histidine in the axial species. PMID- 9414234 TI - The tautomeric state of histidines in myoglobin. AB - 1H-15N HMQC spectra were collected on 15N-labeled sperm whale myoglobin (Mb) to determine the tautomeric state of its histidines in the neutral form. By analyzing metaquoMb and metcyanoMb data sets collected at various pH values, cross-peaks were assigned to the imidazole rings and their patterns interpreted. Of the nine histidines not interacting with the heme in sperm whale myoglobin, it was found that seven (His-12, His-48, His-81, His-82, His-113, His-116, and His 119) are predominantly in the N epsilon2H form with varying degrees of contribution from the Ndelta1 H form. The eighth, His-24, is in the Ndelta1H state as expected from the solid state structure. 13C correlation spectra were collected to probe the state of the ninth residue (His-36). Tentative interpretation of the data through comparison with horse Mb suggested that this ring is predominantly in the Ndelta1H state. In addition, signals were observed from the histidines associated with the heme (His-64, His-93, and His-97) in the 1H-15N HMQC spectra of the metcyano form. In several cases, the tautomeric state of the imidazole ring could not be derived from inspection of the solid state structure. It was noted that hydrogen bonding of the ring was not unambiguously reflected in the nitrogen chemical shift. With the experimentally determined tautomeric state composition in solution, it will be possible to broaden the scope of other studies focused on the electrostatic contribution of histidines to the thermodynamic properties of myoglobin. PMID- 9414235 TI - Temperature dependence of histidine ionization constants in myoglobin. AB - The standard enthalpy of ionization of six titratable histidines in horse metaquomyoglobin was determined by repeating proton NMR titrations as a function of temperature and using the van't Hoff relationship. It was found that deltaH degrees varies between 16 and 37 kJ mol(-1) in the protein, compared with a value of 29 kJ mol(-1) in free histidine. The standard entropy change was evaluated by combining the enthalpy and free energy changes derived from the pKa values. Although the entropy change could not be precisely and accurately obtained by this method, it could be established that it spans a wide range, from -60 to 0 J K(-1) mol(-1), about the value of -23 J K(-1) mol(-1) for the free histidine. The entropy change was used within the framework of enthalpy-entropy compensation to partition the solvation component from the standard thermodynamic quantities for each of the titrating residues. It was shown that the partitioning of the values in the protein is not readily understood in terms of solvent accessibility or electrostatic interactions. The contribution of solvation effects to the temperature response appeared to be significant only in the case of His-119 and His-48. The standard quantities were also used to explore the energetics of proton binding in the native state at temperatures below the onset of thermal denaturation. PMID- 9414236 TI - Effects of molecular crowding on the interaction between DNA and the Escherichia coli regulatory protein TyrR. AB - Fluorescence quenching has been used to measure quantitatively the effects of sucrose and triethylene glycol on the interaction between the Escherichia coli regulatory protein TyrR and a 30-basepair oligonucleotide containing the strong TyrR box of the TyrR operon. It was observed that the apparent binding constant increased in the presence of co-solutes, the dependence of the logarithm of the apparent binding constant on molar concentration being indistinguishable and essentially linear for both co-solutes. This activation of the TyrR oligonucleotide interaction is attributed to thermodynamic nonideality arising from molecular crowding, an interpretation which is supported by the reasonable agreement observed between the experimental extent of reaction enhancement and that predicted on the statistical-mechanical basis of excluded volume. PMID- 9414237 TI - Electrostatic effects on electron-transfer kinetics in the cytochrome f plastocyanin complex. AB - In a complex of two electron-transfer proteins, their redox potentials can be shifted due to changes in the dielectric surroundings and the electrostatic potentials at each center caused by the charged residues of the partner. These effects are dependent on the geometry of the complex. Three different docking configurations (DCs) for intracomplex electron transfer between cytochrome f and plastocyanin were studied, defined by 1) close contact of the positively charged region of cytochrome f and the negatively charged regions of plastocyanin (DC1) and by (2, 3) close contact of the surface regions adjacent to the Fe and Cu redox centers (DC2 and DC3). The equilibrium energetics for electron transfer in DC1-DC3 are the same within approximately +/-0.1 kT. The lower reorganization energy for DC2 results in a slightly lower activation energy for this complex compared with DC1 and DC3. The long heme-copper distance (approximately 24 A) in the DC1 complex drastically decreases electronic coupling and makes this complex much less favorable for electron transfer than DC2 or DC3. DC1-like complexes can only serve as docking intermediates in the pathway toward formation of an electron-transfer-competent complex. Elimination of the four positive charges arising from the lysine residues in the positive patch of cytochrome f, as accomplished by mutagenesis, exerts a negligible effect (approximately 3 mV) on the redox potential difference between cyt f and PC. PMID- 9414238 TI - Fluorescence properties of a new guanosine analog incorporated into small oligonucleotides. AB - The fluorescence properties of 3-methyl-isoxanthopterin (3-MI) incorporated into different oligonucleotides have been determined. This highly fluorescent guanosine analog has its absorption and fluorescence spectra well resolved from those of the normal nucleotides and the aromatic amino acids. The small shifts observed in absorption and fluorescence emission spectra upon incorporation of 3 MI into these oligonucleotides are consistent with a general solvent effect and do not suggest any contribution from the position of the probe from the 5' end, the sequence of nucleotides immediately 5' or 3' to the probe, or the single- or double-stranded nature of the oligomer. However, steady-state and time-resolved fluorescence studies indicate that the presence of a purine immediately 5' or 3' to the probe results in some dynamic but mostly static quenching in the single stranded oligomer. Furthermore, a 3' purine is more effective than a 5' purine, and an adenine appears to be more effective than a guanine for these static quenching interactions. Formation of the double-stranded oligomer leads to an additional loss of quantum yield, which can also be ascribed primarily to static quenching. These results show that this new class of spectrally enhanced fluorescent purine analogs will be able to provide useful information concerning the perturbation of nucleic acid structures. PMID- 9414239 TI - Determination of interspin distances between spin labels attached to insulin: comparison of electron paramagnetic resonance data with the X-ray structure. AB - A method was developed to determine the interspin distances of two or more nitroxide spin labels attached to specific sites in proteins. This method was applied to different conformations of spin-labeled insulins. The electron paramagnetic resonance (EPR) line broadening due to dipolar interaction is determined by fitting simulated EPR powder spectra to experimental data, measured at temperatures below 200 K to freeze the protein motion. The experimental spectra are composed of species with different relative nitroxide orientations and interspin distances because of the flexibility of the spin label side chain and the variety of conformational substates of proteins in frozen solution. Values for the average interspin distance and for the distance distribution width can be determined from the characteristics of the dipolar broadened line shape. The resulting interspin distances determined for crystallized insulins in the R6 and T6 structure agree nicely with structural data obtained by x-ray crystallography and by modeling of the spin-labeled samples. The EPR experiments reveal slight differences between crystal and frozen solution structures of the B chain amino termini in the R6 and T6 states of hexameric insulins. The study of interspin distances between attached spin labels can be applied to obtain structural information on proteins under conditions where other methods like two dimensional nuclear magnetic resonance spectroscopy or x-ray crystallography are not applicable. PMID- 9414240 TI - Investigation of the image contrast of tapping-mode atomic force microscopy using protein-modified cantilever tips. AB - In this work we have designed a simple system to investigate empirically the image contrast of tapping-mode atomic force microscopy (TMAFM). We modified the cantilever tips with protein molecules (bovine serum albumin or goat anti-biotin antibody) and used these protein-modified cantilevers to scan poly-L-lysine films and antibody layers deposited on mica in air under ambient conditions. We also investigated the effects of manipulating the setpoint voltage in this system. It was found that extra topographic features with a patchlike appearance were introduced into the TMAFM images of both the poly-L-lysine and antibody films when scanned with the protein-modified tips, even at initial preset setpoints, and were superimposed on the topography of the samples. The surface coverage of the patchlike features in the TMAFM images changes significantly with the setpoint voltage in a reversible and nonlinear manner. These are believed to arise from the surface indentation of the sample or from the structural deformation of the proteins at the tip induced in TMAFM imaging. Interestingly, it was observed in the experiment that no structural alteration or damage was discernible on the sample surface, even after continuous scanning with the protein-modified tips for a long period of time, with varying setpoint voltage. This study provides experimental evidence that cantilever tips modified with protein molecules or, under certain circumstances, even unmodified tips introduce extra topographical features (i.e., artifacts) and enhance the image contrast of TMAFM imaging of soft materials, which is dependent on their mechanical properties. PMID- 9414241 TI - Introducing phase analysis light scattering for dielectric characterization: measurement of traveling-wave pumping. AB - Phase analysis light scattering (PALS) was applied to characterize a high frequency traveling-wave (TW) micropump. Field strength and frequency characteristics were measured for aqueous solutions up to 40 MHz and conductivities of 16 mS/m. The TW field was generated by an ultramicroelectrode array of intercastellated electrodes, which were driven by square-topped signals. Pumping exhibited one major relaxation peak, which strongly increased for conductivities above 4 mS/m. The conductivity dependence of the peak frequency showed an unexpected nonlinear behavior. Around 20 MHz an additional peak caused by electronic resonance was found. Additional coils or capacitors shifted the resonance peak and allowed us to determine the electronic properties of the array. Analysis of distortions in the pump spectra caused by the harmonic content of the driving signals showed that the pump direction is determined by the traveling direction of the field. For measurement of AC-field-induced particle translations, the advantage of PALS over the commonly used microscopic analysis is that it offers an objective method for statistically significant, computerized registration of extremely slow motions. Thus, for dielectric characterization, low field strengths can be used, which is advantageous not only for analyzing liquid pumping, but also for measuring particle translations induced by dielectrophoresis or TW dielectrophoresis. PMID- 9414242 TI - Localization and molecular interactions of mitoxantrone within living K562 cells as probed by confocal spectral imaging analysis. AB - Studying mechanisms of drug antitumor action is complicated by the lack of noninvasive methods enabling direct monitoring of the state and interactions of the drugs within intact viable cells. Here we present a confocal spectral imaging (CSI) technique as a method of overcoming this problem. We applied this method to the examination of localization and interactions of mitoxantrone (1, 4-dihydroxy 5, 8-bis-[([2-(2-hydroxyethyl)-amino]ethyl)amino]-9,10-anthracenedione dihydrochloride), a potent antitumor drug, in living K562 cells. A two dimensional set of fluorescence spectra of mitoxantrone (MITOX) recorded with micron resolution within a drug-treated cell was analyzed to reveal formation of drug-target complexes and to create the maps of their intracellular distribution. The analysis was based on detailed in vitro modeling of drug-target (DNA, RNA, DNA topoisomerase II) interactions and environmental effects affecting drug fluorescence. MITOX exposed to aqueous intracellular environment, MITOX bound to hydrophobic cellular structures, complexes of MITOX with nucleic acids, as well as the naphtoquinoxaline metabolite of MITOX were simultaneously detected and mapped in K562 cells. These states and complexes are known to be immediately related to the antitumor action of the drug. The results obtained present a basis for the subsequent quantitative analysis of concentration and time-dependent accumulation of free and bound MITOX within different compartments of living cancer cells. PMID- 9414243 TI - Quantitative confocal spectral imaging analysis of mitoxantrone within living K562 cells: intracellular accumulation and distribution of monomers, aggregates, naphtoquinoxaline metabolite, and drug-target complexes. AB - Confocal spectral imaging (CSI) technique was used for quantitative analysis of the uptake, subcellular localization, and characteristics of localized binding and retention of anticancer agent mitoxantrone (MITOX) within human K562 erythroleukemia cells. The CSI technique enables identification of the state and interactions of the drug within the living cells. Utilizing this unique property of the method, intracellular distributions were examined for monomeric MITOX in polar environment, MITOX bound with hydrophobic cellular structures, naphthoquinoxaline metabolite, and nucleic acid-related complexes of MITOX. The features revealed were compared for the cells treated with 2 microM or 10 microM of MITOX for 1 h and correlated to the known data on antitumor action of the drug. MITOX was found to exhibit high tendency to self-aggregation within intracellular media. The aggregates are concluded to be a determinant of long term intracellular retention of the drug and a source of persistent intracellular binding of MITOX. Considerable penetration of MITOX in the hydrophobic cytoskeleton structures as well as growing accumulation of MITOX bound to nucleic acids within the nucleus were found to occur in the cells treated with a high concentration of the drug. These effects may be among the factors stimulating and/or accompanying high-dose mitoxantrone-induced programmed cell death or apoptosis. PMID- 9414244 TI - Delta-opiate DPDPE in magnetically oriented phospholipid micelles: binding and arrangement of aromatic pharmacophores. AB - D-Penicillamine(2,5)-enkephalin (DPDPE) is a potent opioid peptide that exhibits a high selectivity for the delta-opiate receptors. This zwitterionic peptide has been shown, by pulsed-field gradient 1H NMR diffusion studies, to have significant affinity for a zwitterionic phospholipid bilayer. The bilayer lipid is in the form of micelles composed of dihexanoylphosphatidylcholine (DHPC) and dimyristoylphosphatidylcholine (DMPC) mixtures, where the DMPC forms the bilayer structure. At high lipid concentration (25% w/w) these micelles orient in the magnetic field of an NMR spectrometer. The resulting 1H-13C dipolar couplings and chemical shift changes in the natural abundance 13C resonances for the Tyr and Phe aromatic rings were used to characterize the orientations in the bilayer micelles of these two key pharmacophores. PMID- 9414245 TI - Inositol trisphosphate and ryanodine receptors share a common functional Ca2+ pool in cerebellar Purkinje neurons. AB - Changes in the intracellular free calcium concentration ([Ca2+]i) control many important processes in excitable and nonexcitable cells. In cerebellar Purkinje neurons, increases in [Ca2+]i modulate excitability by turning on calcium activated potassium and chloride conductances, and modifying the synaptic efficacy of inhibitory and excitatory inputs to the cell. Calcium release from the intracellular stores plays an important role in the regulation of [Ca2+]i. Purkinje neurons contain both inositol trisphosphate (InsP3) and ryanodine (Ry) receptors. With the exception of the dendritic spines, where only InsP3 receptors are found, InsP3 and Ry receptors are present in the entire cell. The distribution of the two calcium release channels, however, is not uniform, and it has been suggested that InsP3 and Ry receptors use separate Ca2+ pools. The functional properties of InsP3 and Ry Ca2+ pools were investigated by flash photolysis and single-cell microspectrofluorimetry. It was found that depletion of ryanodine-sensitive Ca2+ stores renders InsP3 incapable of releasing more Ca2+ from the stores. Abolishing calcium-induced calcium release by blocking ryanodine receptors with ruthenium red did not have a significant effect on InsP3-evoked Ca2+ release. It is concluded that InsP3 receptors use the same functional Ca2+ pool as that utilized by Ry receptors in Purkinje neurons. PMID- 9414246 TI - Basal intracellular free Mg2+ concentration in smooth muscle cells of guinea pig tenia cecum: intracellular calibration of the fluorescent indicator furaptra. AB - Longitudinal muscle strips dissected from tenia cecum of guinea pig were loaded with the Mg2+ indicator, furaptra, and the relation between the fluorescent ratio signal (R) and cytoplasmic free Mg2+ concentration ([Mg2+]i) was studied in smooth muscle cells at 25 degrees C. After the application of ionophores (4-bromo A23187, monensin, and nigericin), a small immediate offset of R (deltaRjump) was followed by a slow change in R (deltaRslow), which reached a steady level within 2-5 h. The deltaRjump was independent of Mg2+ concentration in solution ([Mg2+]o), and was thought to be unrelated to the change in [Mg2+]i. The direction of the deltaRslow depended on [Mg2+]o with a reversal at approximately 1 mM [Mg2+]o. The intracellular calibration curve was constructed from the steady levels of deltaRslow, and the dissociation constant was 5.4 mM. With the intracellular calibration curve and correction for the deltaRjump, basal [Mg2+], was estimated to be 0.98 +/- 0.05 mM (mean +/- SE, n = 12). When the same calibration was applied to A7r5 cells and rat ventricular myocytes, estimates of basal [Mg2+]i of these cells were 0.74 +/- 0.02 mM (n = 33) and 1.13 +/- 0.06 mM (n = 9), respectively. These results suggest that the basal [Mg2+] level is approximately 1 mM at least in some types of smooth muscle cells, as generally found in striated muscles. PMID- 9414248 TI - Advantages of non-human adenoviruses versus human adenoviruses. PMID- 9414247 TI - Na+ gradient-dependent Mg2+ transport in smooth muscle cells of guinea pig tenia cecum. AB - Thin strips of guinea pig tenia cecum were loaded with the Mg2+ indicator furaptra, and the indicator fluorescence signals measured in Ca2+-free condition were converted to cytoplasmic-free Mg2+ concentration ([Mg2+]i). Lowering the extracellular Na+ concentration ([Na+]o) caused a reversible increase in [Mg2+]i, consistent with the inhibition of Na+ gradient-dependent extrusion of cellular Mg2+ (Na+-Mg2+ exchange). Curve-fitting analysis indicated that the relation between [Na+]o and the rate of rise in [Mg2+], had a Hill coefficient of approximately 3, a [Na+]o at the half-maximal rate of rise of approximately 30 mM, and a maximal rate of 0.16 +/- 0.01 microM/s (mean +/- SE, n = 6). Depolarization with 56 mM K+ shifted the curve slightly toward higher [Na+]o without significantly changing the maximal rate, suggesting that the Na+-Mg2+ exchange was inhibited by depolarization. The maximal rate would correspond to a flux of 0.15-0.4 pmol/cm2/s, if cytoplasmic Mg2+ buffering power (defined as the ratio of the changes in total Mg2+ and free Mg2+ concentrations) is assumed to be 2-5. Ouabain (1-5 microM) increased the intracellular Na+ concentration, as assessed with fluorescence of SBFI (sodium-binding benzofuran isophthalate, a Na+ indicator), and elevated [Mg2+]i. In ouabain-treated preparations, removal of extracellular Na+ rapidly increased [Mg2+]i, with an initial rate of rise roughly proportional to the degree of the Mg2+ load, and, probably, to the Na+ load caused by ouabain. The enhanced rate of rise in [Mg2+]i (up to approximately 1 microM/s) could be attributed to the Mg2+ influx as a result of the reversed Na+ Mg2+ exchange. Our results support the presence of a reversible and possibly electrogenic Na+-Mg2+ exchange in the smooth muscle cells of tenia cecum. PMID- 9414249 TI - Promoter attenuation in gene therapy: causes and remedies. PMID- 9414250 TI - Efficient transduction of mammalian cells by a recombinant baculovirus having the vesicular stomatitis virus G glycoprotein. AB - Baculovirus vectors recently have been shown to be capable of efficient transduction of human hepatoma cells and primary hepatocytes in culture. This paper describes the generation of a novel recombinant baculovirus (VGZ3) in which the vesicular stomatitis virus glycoprotein G (VSV G) is present in the viral envelope. The gene encoding VSV G was inserted into the baculovirus genome under the control of the polyhedrin promoter such that it was expressed at very high levels in infected insect cells but not in mammalian cells. Expression of the lacZ reporter gene was driven by a promoter that is functional in mammalian cells (the Rous sarcoma virus long terminal repeat). We show by Western analysis that VSV G protein was present in purified baculovirus preparations. A VSV G monoclonal antibody blocked transduction of mammalian cells by VGZ3. This virus was morphologically distinct from baculovirus lacking VSV G, with virions adopting an oval rather than rod-shaped morphology. VGZ3 transduced human hepatoma cells in vitro at an efficiency roughly 10-fold greater than baculovirus lacking VSV G (the virus Z4). VGZ3 was also capable of transducing cell lines that could not be transduced efficiently by Z4. We provide evidence that VSV G protein may enhance transduction by increasing the efficiency of escape of baculovirus from intracellular vesicles rather than by increasing cell binding or uptake of the virus. The possible use of this and related baculoviruses in gene therapy is discussed. PMID- 9414251 TI - Promoter attenuation in gene therapy: interferon-gamma and tumor necrosis factor alpha inhibit transgene expression. AB - One of the major limitations to current gene therapy is the low-level and transient vector gene expression due to poorly defined mechanisms, possibly including promoter attenuation or extinction. Because the application of gene therapy vectors in vivo induces cytokine production through specific or nonspecific immune responses, we hypothesized that cytokine-mediated signals may alter vector gene expression. Our data indicate that the cytokines interferon gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) inhibit transgene expression from certain widely used viral promoters/enhancers (cytomegalovirus, Rous sarcoma virus, simian virus 40, Moloney murine leukemia virus long terminal repeat) delivered by adenoviral, retroviral or plasmid vectors in vitro. A constitutive cellular promoter (beta-actin) is less sensitive to these cytokine effects. Inhibition is at the mRNA level and cytokines do not cause vector DNA degradation, inhibit total cellular protein synthesis, or kill infected/transfected cells. Administration of neutralizing anti-IFN-gamma monoclonal antibody results in enhanced transgene expression in vivo. Thus, standard gene therapy vectors in current use may be improved by altering cytokine responsive regulatory elements. Determination of the mechanisms involved in cytokine-regulated vector gene expression may improve the understanding of the cellular disposition of vectors for gene transfer and gene therapy. PMID- 9414252 TI - Semliki Forest virus-mediated production of retroviral vector RNA in retroviral packaging cells. AB - Retroviral vectors are efficient tools for gene transfer studies. Their major advantage is that they can permanently integrate the transgene into the target cell's genome. However, because of the compulsory nuclear expression phase of their life cycle, it can be difficult for retroviruses to carry complex expression cassettes. In a attempt to mimic the structural features of most eukaryotic genes and obtain a potentially self-amplifying system for retrovirus production, we tested the feasibility of Semliki Forest virus (SFV) expression to mediate cytoplasmic synthesis of retrovirus vector RNA. An equivalent of a retrovirus virion RNA (retrovirus vector cassette, RVC) was cloned under the SFV 26S promoter, and full-length chimeric SFV-RVC RNA was produced in vitro. This RNA was introduced into retrovirus packaging cells, either via electroporation or transduction in SFV virions, and supernatants were analyzed for the presence of biologically active retroviruses. We demonstrate that this strategy can be used for cytoplasmic retrovirus production. The resulting viral particles are fully functional; they can transduce target cells, undergo reverse transcription, and integrate into genomic DNA. We also demonstrate that the SFV virion-based RVC delivery into packaging cells can yield high transient titers, in this case more than 10(5) G418R cfu/ml. This study shows that a simple, one-plasmid, heterologous viral RNA production system can be used to create functional retroviral RNA outside the cell nucleus. PMID- 9414253 TI - Regression of experimental brain tumors with 6-thioxanthine and Escherichia coli gpt gene therapy. AB - The identification of transgenes with antitumor activity is critical to the development of gene therapy of cancer. Retrovirus-mediated transfer of the Escherichia coli gpt gene into rat C6 glioma cells without subsequent selection still inhibited the proliferation of this mixed polyclonal population upon addition of the prodrug, 6-thioxanthine, with an ID50 of 4.1 microM, whereas parental C6 cells were not affected at a concentration of 500 microM. In a time course assay, effects of the prodrug on the mixed polyclonal cell proliferation required at least 10 days of exposure. In mixed co-cultures, a bystander effect was not present over the first 4 days of prodrug exposure, but required trypsinization of the co-cultures and replating at lower densities. This "modified" bystander assay thus revealed a 50% decrease in C6 cell proliferation, even when the initial ratio of gpt-expressing to parental C6 cells was as low as 1:19. In a nude mouse model of subcutaneous tumors, co-grafts of C6 glioma and gpt-retrovirus producer cells displayed retarded growth upon exposure to 6 thioxanthine (6-TX). In a nude mouse model of intracerebral tumors, grafting of the gpt-retrovirus producer cells leads to an 80% reduction in intracerebral tumor volumes after 6-TX treatment. This reduction results in a 28% increase in the mean time of survival of animals that harbor intracerebral tumors (p < 0.0005). These antitumor effects indicate that the gpt/6-TX enzyme/prodrug pair is a promising alternative to the thymidine kinase gene and ganciclovir combination in the gene therapy of cancer. PMID- 9414254 TI - Therapeutic efficiency and safety of a second-generation replication-conditional HSV1 vector for brain tumor gene therapy. AB - A second-generation replication-conditional herpes simplex virus type 1 (HSV) vector defective for both ribonucleotide reductase (RR) and the neurovirulence factor gamma34.5 was generated and tested for therapeutic safety and efficiency in two different experimental brain tumor models. In culture, cytotoxic activity of this double mutant HSV vector, MGH-1, for 9L gliosarcoma cells was similar to that of the HSV mutant, R3616, which is defective only for gamma34.5, but was significantly weaker than that of the HSV mutant hrR3, which is defective only for RR. The diminished tumoricidal effect of the gamma34.5 mutants could be accounted for by their reduced ability to replicate in 9L cells. The MGH-1 vector did not achieve significant prolongation of survival in vivo in the syngeneic 9L rat gliosarcoma model for either single brain tumor focus or multiple intracerebral and leptomeningeal tumors, when the vector was applied intratumorally or intrathecally, respectively, and with or without subsequent ganciclovir (GCV) treatment. In identical 9L brain tumor models with single and multiple foci, application of hrR3 with or without GCV was previously shown to result in marked long-term survival. Contrary to the findings with intrathecal injection of hrR3, no vector-related mortality was observed in any animals treated with MGH-1. Thus, in these rat brain tumor models, the double mutant, replication-conditional HSV vector MGH-1 showed a higher therapeutic safety than the RR-minus vector, hrR3, but had clearly decreased therapeutic efficiency compared to hrR3. The development of new HSV vectors for brain tumor gene therapy will require a balance between maximizing therapeutic efficacy and minimizing toxicity to the brain. Standardized application in brain tumor models as presented here will help to screen new HSV vectors for these requirements. PMID- 9414255 TI - Comparison of the protection of cells from antifolates by transduced human dihydrofolate reductase mutants. AB - Retroviral transduction of antifolate-resistant variants of human dihydrofolate reductase (hDHFR) into cells can increase their resistance to the cytotoxic effects of these drugs. We evaluated the ability of wild-type hDHFR and 20 mutant enzymes (13 with single-amino acid substitutions, 7 with two substitutions) to prevent growth inhibition in antifolate-treated CCRF-CEM cells. The wild-type enzyme and all of the variants significantly protected transduced cells from trimetrexate (TMTX)-induced growth inhibition. However, only half of the variants conferred more protection than does the wild-type enzyme. For the variants tested, the observed protective effect was higher for TMTX than for methotrexate (< or =7.5-fold increased resistance), piritrexim (< or =16-fold), and edatrexate (negligible). Transduction of the variants L22Y-F31S and L22Y-F31R led to the greatest protection against TMTX (approximately 200-fold). Protection from loss of cell viability was similar to protection from growth inhibition. The protection associated with a particular mutant hDHFR did not result from the level of expression: Efficient protection resulted from low affinity of the variant for antifolates, reasonable catalytic activity, and good thermal stability. Clones isolated from a polyclonal population of transduced cells varied by as much as 30-fold in their resistance to TMTX, the resistance differences depending on hDHFR expression levels. PMID- 9414256 TI - Efficient gene transfer in primitive CD34+/CD38lo human bone marrow cells reselected after long-term exposure to GALV-pseudotyped retroviral vector. AB - Successful retroviral gene transfer into human hematopoietic stem cells was demonstrated in preliminary clinical trials at low efficiency. We have shown previously that gene transfer into committed hematopoietic progenitor cells is more efficient using a gibbon ape leukemia virus (GALV)-pseudotyped retroviral vector instead of an amphotropic retroviral vector. Here, we have conducted a systematic study of human hematopoietic progenitor cells after extended transduction with a GALV-pseudotyped retroviral vector. CD34+/CD38lo Cells were transduced for 5 days and reselected according to phenotype after culture and analyzed for cell cycle status, long-term culture-initiating cell (LTC-IC) activity, and gene transfer. Reselection of rare, very primitive progenitor cells was successful. Equal to fresh CD34+/CD38lo cells, >90% of reselected CD34+/CD38lo cells were in G0/G1. CD34+/CD38lo reselection enriched for LTC-IC (10-fold), as compared to freshly isolated CD34+/CD38lo cells with excellent specificity (82.7% of total LTC-IC were recovered in the reselected CD34+/CD38lo population) and recovery (62% of initial LTC-IC number in CD34+/CD38lo cells were recovered in the reselected fraction after transduction). Gene transfer into primitive progenitor cells was efficient with 50.5% G418-resistant LTC-IC colonies and more than 40 copies of vector provirus detectable per 100 nuclei of CD34+/CD38lo cells. To our knowledge, this is the first systematic analysis of phenotype, function, and cell cycle demonstrating retroviral gene transfer into rare, very primitive human hematopoietic progenitor cells. The chosen strategy should be of considerable value for analyzing and improving gene therapy of the hematopoietic system. PMID- 9414257 TI - A family of bicistronic vectors to enhance both local and systemic antitumor effects of HSVtk or cytokine expression in a murine melanoma model. AB - The herpes simplex virus-thymidine kinase/ganciclovir (HSVtk/GCV) system produces both direct and immune-mediated tumor cell killing. Here, we compare the efficacy of HSVtk/GCV with cytokines, alone and in combination, on the tumorigenicity and immunogenicity of B16 cells. With respect to single gene modifications, only HSVtk/GCV, or high-level interleukin-2 (IL-2) secretion, completely prevented tumor growth, whereas granulocyte-macrophage colony-stimulating factor (GM-CSF) generated the best levels of long-term systemic protection. To augment both local killing and immune activation, we constructed bicistronic constructs that express HSVtk and a cytokine within the same cell. Co-expression of HSVtk with IL-2 or GM CSF enhanced the local antitumor activity of any gene alone. In a tumor prevention model, HSVtk killing, in an environment preprimed with GM-CSF, generated the best long-term immune protection. However, in a short-term therapy model, continued IL-2 expression was most effective against 3-day established tumors. This probably reflects differences in the activities of IL-2 and GM-CSF in generating short-term, nonspecific immune stimulation compared to long-term immunological memory, respectively. As a prelude to in vivo delivery experiments, we also demonstrated that these bicistronic cassettes can be packaged normally into retroviral (5 x 10(5) virus/ml from pooled populations) and adenoviral vectors (5 x 10(9) virus/ml) and function as predicted within virally infected cells. This family of bicistronic vectors can be used to stimulate synergy between suicide and cytokine genes, overcomes the problems of delivering two genes on separate vectors, and should allow easier preparation of vectors for the delivery of multiple genes to patients' tumor cells. PMID- 9414259 TI - Overexpression of adenovirus-encoded transgenes from the cytomegalovirus immediate early promoter in irradiated tumor cells. AB - Efficient expression of therapeutic genes in irradiated tumor cells would facilitate the conversion of a malignant tumor nodule into a cancer vaccine in situ. We reported previously that transgene expression from an adenoviral vector could be markedly enhanced by treating transduced tumor cells with butyrate. In this study, we demonstrated that a similar butyrate effect could be achieved in irradiated tumor cells. In addition, irradiating cells at doses of 2-40 Gy prior to transduction could also amplify recombinant adenoviral transgene products in a cell-type-specific manner. This suggests that adenovirus-mediated gene therapy, radiation therapy, and butyrate-mediated cancer therapy may potentially be formulated into one synergistic protocol for cancer treatment. PMID- 9414258 TI - A recombinant E1-deleted canine adenoviral vector capable of transduction and expression of a transgene in human-derived cells and in vivo. AB - Human adenovirus (HAV) serotypes 2 and 5 are commonly used as vector backbones for adenovirus-mediated gene transfer. However, HAVs were chosen as a backbone for the vectors for historical reasons and have a number of significant disadvantages when used as a shuttle for gene transfer in humans. As an initial trial to circumvent some of the shortcomings of HAV vectors, we have produced an E1-deleted canine adenovirus type 2 (CAV-2) vector for gene transfer. Initially, we demonstrated that CAV-2 undergoes an abortive viral cycle in a wide range of human-derived cell lines. Second, we assayed human sera containing HAV-5 neutralizing antibodies for their ability to inhibit CAV-2-induced plaques on permissive cells. In the cohort tested, our data demonstrate that the humoral response directed against HAV-5 does not inhibit CAV-2 plaque formation in the majority of cases. Canine cell lines expressing the E1 region of CAV-2 were generated and characterized. A recombinant CAV vector (CAVRSVbetagal) deleted in the E1 region and harboring lacZ was constructed. We show that CAVRSVbetagal is able to transduce and direct expression of the transgene in vitro in a variety of mammalian cells, most notably primary human-derived cells. In addition, gene transfer is demonstrated in vivo using chick embryos. PMID- 9414260 TI - Retrovirus-mediated gene transfer of ornithine-delta-aminotransferase into keratinocytes from gyrate atrophy patients. AB - Gyrate atrophy is a progressive blindness associated with deficiency of ornithine aminotransferase (OAT). The strategy of using an autologous keratinocyte graft, modified to express high levels of OAT as an ornithine-catabolizing skin-based enzyme sink, is investigated. Two OAT-containing retroviral vectors were constructed with or without a resistance gene. When packaged in a retroviral vector particle generated with the gibbon ape leukemia (GALV) virus envelope (PG13), these vectors could readily transduce >50% of target keratinocytes. The transduced keratinocytes in culture expressed up to 75-fold more OAT than normal control keratinocytes and these gene-modified cells extracted [14C]ornithine more efficiently than controls. The vector prepared without neo transduced cells more efficiently and led to higher levels of OAT expression than the neo-containing vector. Ornithine catabolism was maintained at high levels when the transduced patient keratinocytes were differentiated in vitro as a multilayered cutaneous organoid. PMID- 9414261 TI - Development and characterization of cationic liposomes conjugated with HVJ (Sendai virus): reciprocal effect of cationic lipid for in vitro and in vivo gene transfer. AB - Today, nonviral gene transfer vectors attract more attention as a therapeutic strategy for human diseases, because viral vectors such as adenoviral and herpes viral vectors have been proven to have problems, especially in immunogenicity and cytotoxicity. However, the main limitation of nonviral vectors has been low efficiency of gene expression. To overcome this defect, we have developed a new class of transfection vehicles, HVJ-cationic liposomes. The use of the cationic lipid DC-cholesterol facilitates efficient entrapment of negatively charged macromolecules (plasmid DNA, oligodeoxynucleotides, and proteins) and efficient interaction with negatively charged plasma membranes of cultured cells in vitro. Moreover, the fusogenic envelope proteins of hemagglutinating virus of Japan (HVJ) enhance delivery of the enclosed materials into cells. Using firefly luciferase as a marker, we optimized the liposome formula. As a result, we have succeeded in obtaining 100-800 times higher gene expression in vitro than with the conventional HVJ-anionic liposomes. However, in vivo gene transfer experiments have revealed that the use of cationic lipid instead of anionic lipid reduced the transgene expression dramatically in organs such as muscle and liver. We further discovered that the use of anionic liposomes with a viral-mimic king lipid composition increased transfection efficiency by several times in vivo. We conclude that the alternative usage of transfer vectors, for example, HVJ-anionic liposomes for in vivo delivery to extended areas of organs and HVJ-cationic liposomes for in vitro delivery (and possibly for in vivo delivery to a restricted area of organs), is of significance. PMID- 9414263 TI - A new year greeting from a new editor PMID- 9414262 TI - Retrovirus-mediated transfer of the cDNA for human glucocerebrosidase into peripheral blood repopulating cells of patients with Gaucher's disease. PMID- 9414265 TI - Lack of interferon consensus sequence binding protein (ICSBP) transcripts in human myeloid leukemias. AB - Interferon consensus sequence binding protein (ICSBP) was first identified as a transcription factor of the interferon (IFN) regulatory factor family (IRF) which regulates expression of IFN-dependent genes by binding to DNA at specific sites, IFN-stimulated responsive elements. Analysis of ICSBP-deficient mice showed hematologic alterations similar to chronic myelogenous leukemia (CML) in humans and suggested a novel role for ICSBP in regulating proliferation and differentiation of hematopoietic progenitor cells. Here we show that ICSBP-mRNA expression is impaired in human myeloid leukemias: 27 of 34 CML patients (79%) and 21 of 32 patients with acute myeloid leukemia (AML) (66%) showed very low or absent transcript numbers of ICSBP. In contrast, only 2 of 33 normal volunteers (6%) showed low transcription of ICSBP (P < . 0001 both for CML and AML values). The lack of expression was not associated with lack of lymphatic cells, which normally have been shown to express ICSBP at the highest level. More detailed analysis showed an absence of ICSBP-mRNA also in sorted B cells derived from CML patients. To analyze whether ICSBP may be induced in leukemic cells, ex vivo experiments using a known inducer of ICSBP, IFN-gamma, were performed. Ex vivo treatment of primary CML cells using IFN-gamma resulted in induction of ICSBP transcripts. Furthermore, samples of CML patients during IFN-alpha treatment were analyzed. In 11 of 12 CML patients ICSBP-mRNA was inducible upon in vivo treatment with IFN-alpha, but decreased with progression of CML. Stable transfection of K-562 cell line with ICSBP led to no difference in bcr-abl expression in vitro, although two patients showed an inverse correlation between bcr-abl and ICSBP in vivo. These data suggest that lack of ICSBP may have an important role also in human myeloid leukemogenesis. PMID- 9414266 TI - Successful peripheral T-lymphocyte-directed gene transfer for a patient with severe combined immune deficiency caused by adenosine deaminase deficiency. AB - Ten patients with adenosine deaminase deficiency (ADA-) have been enrolled in gene therapy clinical trials since the first patient was treated in September 1990. We describe a Japanese ADA- severe combined immune deficiency (SCID) patient who has received periodic infusions of genetically modified autologous T lymphocytes transduced with the human ADA cDNA containing retroviral vector LASN. The percentage of peripheral blood lymphocytes carrying the transduced ADA gene has remained stable at 10% to 20% during the 12 months since the fourth infusion. ADA enzyme activity in the patient's circulating T cells, which was only marginally detected before gene transfer, increased to levels comparable to those of a heterozygous carrier individual and was associated with increased T lymphocyte counts and improvement of the patient's immune function. The results obtained in this trial are in agreement with previously published observations and support the usefulness of T lymphocyte-directed gene transfer in the treatment of ADA-SCID. PMID- 9414267 TI - Multilineage hematopoietic recovery by a single injection of pegylated recombinant human megakaryocyte growth and development factor in myelosuppressed mice. AB - Previous studies have shown that daily multiple administration of pegylated recombinant human megakaryocyte growth and development factor (PEG-rHuMGDF) markedly stimulates thrombopoiesis and effectively ameliorates thrombocytopenia, and in most cases anemia and neutropenia, in myelosuppressed animals. In this study, we evaluated the effects of a single intravenous injection of PEG-rHuMGDF on hematopoietic recovery after sublethal total-body irradiation in mice. A single injection of PEG-rHuMGDF (1 to 640 microg/kg) 1 hour after irradiation accelerated platelet, red blood cell (RBC), and white blood cell (WBC) recovery in a dose-dependent fashion. In the bone marrow of vehicle-treated mice, megakaryocytic, erythroid, and myeloid progenitors, as well as day 12 colony forming unit-spleen (CFU-S), were dramatically decreased much earlier than the nadirs of peripheral blood cells, whereas megakaryocytes were modestly decreased. Treatment with PEG-rHuMGDF (80 microg/kg, an optimal dose) 1 hour after irradiation resulted in more rapid recovery of these four hematopoietic progenitors and also significantly facilitated megakaryocyte recovery. In addition, the same PEG-rHuMGDF administration schedule expanded bone marrow cells capable of rescuing lethally irradiated recipient mice. As the interval between irradiation and PEG-rHuMGDF treatment was longer, its effects on hematopoietic recovery were attenuated. In contrast to the effects of PEG-rHuMGDF, a single injection of recombinant human granulocyte colony-stimulating factor (rhG-CSF) 1 hour after irradiation exclusively accelerated WBC recovery, but only to a similar extent as PEG-rHuMGDF (80 microg/kg) treatment even when rhG-CSF doses were escalated to 1,000 microg/kg. This appeared related to different pharmacokinetics of these two factors after a single injection in irradiated mice. The concentrations of PEG-rHuMGDF after injection persisted in the plasma for a longer time compared with rhG-CSF. These results indicate that a single injection of PEG-rHuMGDF at an early time after irradiation is able to effectively improve thrombocytopenia, anemia, and leukopenia with concomitant accelerated recovery of both primitive and committed hematopoietic progenitors in irradiated mice. Our data also show that compared with the rhG-CSF shown to exert multilineage effects on hematopoiesis, PEG-rHuMGDF has more wide-ranging effects on peripheral blood cell recovery. PMID- 9414268 TI - c-Cbl is tyrosine-phosphorylated by interleukin-4 and enhances mitogenic and survival signals of interleukin-4 receptor by linking with the phosphatidylinositol 3'-kinase pathway. AB - Interleukin-4 (IL-4) is a cytokine that induces both proliferation and differentiation and suppresses apoptosis of B cells. Although IL-4 has been shown to activate the phosphatidylinositol 3' (PI3)-kinase pathway, the role of PI3 kinase in the IL-4 receptor (IL-4R) signaling remains unclear. In this study, we demonstrated that c-Cbl proto-oncogene product is inducibly phosphorylated on tyrosine residues and is associated with the p85 subunit of PI3-kinase by IL-4 stimulation. Overexpression of c-Cbl enhances the PI3-kinase activity and, at the same time, mitogenic activity and survival of cells in the presence of IL-4. However, these effects of c-Cbl were abolished by wortmannin, a specific inhibitor for the PI3 kinase pathway, or by a point mutation at tyrosine 731 of c Cbl, which is a major binding site for p85. These results indicate that c-Cbl plays a role in linking IL-4R with the PI3 kinase pathway and thus enhancing the mitogenic and survival signals. PMID- 9414269 TI - A truncated isoform of the human beta chain common to the receptors for granulocyte-macrophage colony-stimulating factor, interleukin-3 (IL-3), and IL-5 with increased mRNA expression in some patients with acute leukemia. AB - We report here a naturally occurring isoform of the human beta chain common to the receptors for granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-3 (IL-3), and IL-5 (GMRbetaC) with a truncated intracytoplasmic tail caused by deletion of a 104-bp exon in the membrane-proximal region of the chain. This beta intracytoplasmic truncated chain (betaIT) has a predicted tail of 46 amino acids, instead of 432 for betaC, with 23 amino acids in common with betaC and then a new sequence of 23 amino acids. In primary myeloid cells, betaIT comprised approximately 20% of the total beta chain message, but was increased up to 90% of total in blast cells from a significant proportion of patients with acute leukemia. Specific anti-betaIT antibodies demonstrated its presence in primary myeloid cells and cell lines. Coexpression of betaIT converted low affinity GMRalpha chains (KD 2.5 nmol/L) to higher-affinity alphabeta complexes (KD 200 pmol/L). These could bind JAK2 that was tyrosine-phosphorylated by stimulation with GM-CSF. betaIT did not support GM-CSF-induced proliferation when cotransfected with GMRalpha into CTLL-2 cells. Therefore, it may interfere with the signal-transducing properties of the betaC chain and play a role in the pathogenesis of leukemia. PMID- 9414270 TI - Membrane translocation of 15-lipoxygenase in hematopoietic cells is calcium dependent and activates the oxygenase activity of the enzyme. AB - Mammalian 15-lipoxygenases, which have been implicated in the differentiation of hematopoietic cells are commonly regarded as cytosolic enzymes. Studying the interaction of the purified rabbit reticulocyte 15-lipoxygenase with various types of biomembranes, we found that the enzyme binds to biomembranes when calcium is present in the incubation mixture. Under these conditions, an oxidation of the membrane lipids was observed. The membrane binding was reversible and led to an increase in the fatty acid oxygenase activity of the enzyme. To find out whether such a membrane binding also occurs in vivo, we investigated the intracellular localization of the enzyme in stimulated and resting hematopoietic cells by immunoelectron microscopy, cell fractionation studies and activity assays. In rabbit reticulocytes, the 15-lipoxygenase was localized in the cytosol, but also bound to intracellular membranes. This membrane binding was also reversible and the detection of specific lipoxygenase products in the membrane lipids indicated the in vivo activity of the enzyme on endogenous substrates. Immunoelectron microscopy showed that in interleukin-4 treated monocytes, the 15-lipoxygenase was localized in the cytosol, but also at the inner side of the plasma membrane and at the cytosolic side of intracellular vesicles. Here again, cell fractionation studies confirmed the in vivo membrane binding of the enzyme. In human eosinophils, which constitutively express the 15 lipoxygenase, the membrane bound share of the enzyme was augmented when the cells were stimulated with calcium ionophore. Only under these conditions, specific lipoxygenase products were detected in the membrane lipids. These data suggest that in hematopoietic cells the cytosolic 15-lipoxygenase translocates reversibly to the cellular membranes. This translocation, which increases the fatty acid oxygenase activity of the enzyme, is calcium-dependent, but may not require a special docking protein. PMID- 9414271 TI - Impaired ability of bone marrow stromal cells to support B-lymphopoiesis with age. AB - B-lymphopoiesis decreases with age. We studied how aging affects bone marrow stromal cells, because they provide the growth factors and cell contacts required for B-lymphopoiesis. No differences were noted in the cell-surface phenotype of young and old primary-cultured stromal cells. Fluorescence-activated cell sorter purified stromal cells from old mice were deficient in the ability to support the proliferation of interleukin-7 (IL-7)-specific B-lymphoid cell lines. The kinetics of this response indicated that IL-7 was not immediately available from stromal cells of either age and was further delayed on aged stromal cells. The levels of IL-7 protein within stromal cells were equivalent between young and old animals, suggesting that the production of IL-7 was not altered by aging. Negligible amounts of IL-7 were found either freely secreted or in the extracellular matrix of cultures of young and old marrow. Contact between the lymphoid cells and the primary stromal cells was required for detectable proliferation, suggesting that cell contact was required for the release of IL-7. We propose that stromal cells regulate B-lymphopoiesis by limiting the amount of IL-7 available to the developing precursors. Therefore, we conclude that the age related decrease in the function of bone marrow stromal cells is related to the impaired release of IL-7. PMID- 9414272 TI - Transient thrombocytopenia produced by administration of macrophage colony stimulating factor: investigations of the mechanism. AB - Administration of macrophage colony-stimulating factor (M-CSF) to mice (2 to 8 mg/kg/d x 5d) produced dose-dependent thrombocytopenia, which reached its nadir on days 4 to 5, followed by rapid recovery. Surprisingly, when administration of M-CSF was prolonged, the thrombocytopenia completely resolved, despite continued treatment. Splenectomy did not prevent the thrombocytopenia. Readministration of M-CSF after various intervals continued to produce the thrombocytopenic effect, even after 35 days. Measurements of Meg-CFC and megakaryocyte ploidy during the periods of M-CSF treatment and recovery of normal platelet levels showed no evidence of bone marrow suppression. Platelet survival was markedly decreased after 5 days of M-CSF (at the platelet count nadir) and after 9 days of continued M-CSF treatment, when the platelet count had returned to normal. Platelets from M CSF-treated donors demonstrated normal survival when transfused into normal recipients. We concluded that thrombocytopenia produced by M-CSF was not due to suppression of thrombopoiesis, but to increased activity of the monocyte/macrophage system, which caused shortened platelet survival, and that subsequently, increased platelet production compensated for ongoing platelet destruction and resulted in normal platelet levels. PMID- 9414264 TI - Multiple myeloma: increasing evidence for a multistep transformation process. PMID- 9414273 TI - In vitro behavior of hematopoietic progenitor cells under the influence of chemoattractants: stromal cell-derived factor-1, steel factor, and the bone marrow environment. AB - How multiple chemoattractants cooperate in directing the migration of hematopoietic progenitor cells (HPC) for homing and peripheral blood mobilization has not yet been established. We report here the behavior of HPC under the influence of two different chemoattractants, stromal cell-derived factor (SDF)-1 and steel factor (SLF), and the chemotactic nature of the bone marrow (BM) environment using a two-chamber in vitro migration system. Various formulae were adopted to quantitate these effects. Based on these quantitations, SDF-1 showed only chemotactic activity, while SLF showed both chemotactic and chemokinetic activities on factor-dependent MO7e cells. SLF, like SDF-1, attracted human HPC from a population of CD34+ cells and induced actin polymerization in MO7e cells. SLF and SDF-1 cooperated in attracting MO7e cells, as well as cord blood (CB) and BM CD34+ cells. A negative concentration gradient of SLF and SDF-1, formed by the presence of chemoattractants in the upper chamber, showed potent inhibitory effects on MO7e cell migration induced by either of these chemoattractants in the lower chamber, and SDF-1 and SLF were synergistic in mobilizing cells to the lower chamber from this negative chemoattractant gradient. Plasma obtained from BM aspirates, but not CB or peripheral blood, showed strong chemotactic effects on BM and CB CD34+ cells, and an inhibitory effect in a negative gradient on SDF 1-dependent CD34+ cell migration. These in vitro migration experiments suggest that chemoattractants such as SDF-1 and SLF with other unidentified BM chemoattractants may be involved cooperatively in the migration of HPC to the BM and in preventing spontaneous mobilization of HPC out of the BM. PMID- 9414274 TI - Stroma-contact prevents loss of hematopoietic stem cell quality during ex vivo expansion of CD34+ mobilized peripheral blood stem cells. AB - Stroma-supported long-term cultures (LTC) allow estimation of stem cell quality by simultaneous enumeration of hematopoietic stem cell (HSC) frequencies in a graft using the cobblestone area forming cell (CAFC) assay, and the ability of the graft to generate progenitors in flask LTC (LTC-CFC). We have recently observed that the number and quality of mobilized peripheral blood stem cells (PBSC) was low in patients having received multiple rounds of chemotherapy. Moreover, grafts with low numbers of HSC and poor HSC quality had a high probability to cause graft failure upon their autologous infusion. Because ex vivo culture of stem cells has been suggested to present an attractive tool to improve hematological recovery or reduce graft size, we have studied the possibility that such propagation may affect stem cell quality. In order to do so, we have assessed the recovery of different stem cell subsets in CD34+ PBSC after a 7-day serum-free liquid culture using CAFC and LTC-CFC assays. A numerical expansion of stem cell subsets was observed in the presence of interleukin-3 (IL-3), stem cell factor, and IL-6, while stroma-contact, stromal soluble factors, or combined addition of FLT3-ligand and thrombopoietin improved this parameter. In contrast, ex vivo culture severely reduced the ability of the graft to produce progenitors in LTC while stromal soluble factors partly abrogated this quality loss. The best conservation of graft quality was observed when the PBSC were cultured in stroma-contact. These data suggest that ex vivo propagation of PBSC may allow numerical expansion of various stem cell subsets, however, at the expense of their quality. In addition, cytokine-driven PBSC cultures require stroma for optimal maintenance of graft quality. PMID- 9414275 TI - Purine nucleotides induce regulated secretion of von Willebrand factor: involvement of cytosolic Ca2+ and cyclic adenosine monophosphate-dependent signaling in endothelial exocytosis. AB - von Willebrand factor (vWF) is stored and released from endothelial secretory granules called Weibel-Palade (WP) bodies. Acute release can be induced by thrombin, histamine, and other mediators of thrombosis or inflammation. Their effect is thought to be mediated by an increase in intracellular free calcium ([Ca2+]i). Purine nucleotides such as adenosine triphosphate (ATP) and adenosine diphosphate (ADP) are released from platelet dense granules and from ischemic tissues and are important regulators of platelet function and vascular tone. In the present study, we investigated whether they could also induce exocytosis from cultured endothelial cells. ATP (1 to 100 micromol/L) induced a dose-related increase in vWF release, with a 2.3-fold maximal increase after 30 minutes. Similar responses were observed with ADP. ATP induced calcium mobilization from intracellular stores, an effect mimicked by 2-methylthio-ATP, a selective agonist for P2y receptors. However, 2-methylthio-ATP-induced vWF release was only 43% of the ATP response. ATP-induced vWF release was also associated with a twofold increase in cellular cyclic adenosine monophosphate (cAMP) content, and was potentiated by 3-isobutyl-1-methylxanthine ([IBMX] added to increase cAMP levels by blocking cellular phosphodiesterases) and 8-bromo-cAMP and inhibited by more than 50% by Rp-8-CPT-cAMPS, a competitive protein kinase A inhibitor. Adenosine but not 2-methylthio-ATP mimicked the ATP-induced increase in cAMP. ATP-induced vWF release was partly inhibited by adenosine deaminase, which degrades adenosine generated from ATP in the incubation medium. Adenosine (1 to 100 micromol/L) failed to induce vWF release, but potentiated the secretory response to 2 methylthio-ATP and thrombin without modifying the calcium response to these agents. Our results suggest that ATP/ADP can induce vWF release from endothelial cells via dual activation of P2y and adenosine A2 receptors. ATP/ADP-induced exocytosis could be involved in the regulation of thrombus formation and ischemia reperfusion injuries. Further, we provide evidence that a receptor-mediated increase in cellular cAMP can potentiate the secretory response to calcium mobilizing agents. PMID- 9414276 TI - HRG Tokushima: molecular and cellular characterization of histidine-rich glycoprotein (HRG) deficiency. AB - Previously, we found the first congenital deficiency of histidine-rich glycoprotein (HRG) in a Japanese woman with thrombosis. To elucidate the genetic basis of this deficiency, we first performed Southern blot analysis and found no gross deletion or insertion in the proband's HRG gene. We then examined the nucleotide sequences of all seven exons of the proband's HRG gene. A single nucleotide substitution, G to A at nucleotide position 429, which mutates Gly85 to Glu in the first cystatin-like domain, was found in exon 3 in 13 of 22 amplified clones. This mutation generates a unique Taq I site. Exon 3 was amplified from the proband, her family members, and 50 unrelated normal Japanese individuals, and Taq I fragmentation was examined. Fragmentation of exon 3 was observed in one allele of the genes from the proband and the family members who also have decreased plasma levels of HRG. Fifty unrelated normal Japanese individuals had a normal HRG gene, indicating that the G to A mutation is not a common polymorphism. To elucidate the identified mutation as a cause for the secretion defect of HRG in the proband's plasma, we constructed and transiently expressed the recombinant Tokushima-type HRG mutant (Gly85 to Glu) in baby hamster kidney (BHK) cells, and examined an intracellular event of the mutant protein. The results showed that only about 20% of the Tokushima-type HRG was secreted into the culture medium, and intracellular degradation of the mutant was observed. Thus, the present study strongly suggests that the HRG deficiency is caused by intracellular degradation of the Gly85 to Glu mutant of HRG in the proband. PMID- 9414277 TI - Platelet-derived interleukin-1 induces cytokine production, but not proliferation of human vascular smooth muscle cells. AB - During vascular injury, such as observed in atherosclerosis, restenosis, vasculitides, transplantation, or sepsis, vascular smooth muscle cells (SMC) can be exposed to platelets or platelet products. Under these conditions proliferation or cytokine production of SMC stimulated by platelets or platelet products may contribute to regulation of vascular pathogenesis. Thus, we investigated interleukin-6 (IL-6) and IL-8 production as well as proliferation of SMC in response to platelets or platelet lysates. Platelets not already preactivated by thrombin induced IL-6 (10- to 50-fold) or IL-8 production of unstimulated SMC in a cell number dependent fashion. Preactivation of platelets with thrombin potently increased the platelet-mediated IL-6 (50- to 1,000-fold) and IL-8 production of SMC. Hirudin specifically inhibited the activation of platelets with thrombin. Isolated platelets cultured in the absence of SMC did not contain detectable IL-6 or IL-8. Prestimulation (4 hours) of SMC with pathophysiologically relevant substances (lipopolysaccharide [LPS], tumor necrosis factor-alpha [TNF-alpha], or IL-1alpha) further increased the platelet induced cytokine production. The platelet-derived SMC stimulatory activity was IL 1, since IL-1 receptor antagonist (IL-1-Ra) inhibited the platelet-induced cytokine production of SMC. Anti-platelet-derived growth factor (PDGF)-antibody did not further reduce this activity. Thrombin itself stimulated expression of IL 6 and IL-8 to some degree and induced IL-6 production of SMC synergistically with IL-1. Platelets also induced proliferation of SMC, however, anti-PDGF antibodies, rather than IL-1-Ra blocked this response. These data show that platelet-derived IL-1 stimulates cytokine production of vascular smooth muscle cells, indicating that platelet-derived IL-1 may contribute to regulation of local pathogenesis in the vessel wall by activation of the cytokine regulatory network. PMID- 9414278 TI - Factor VII deficiency caused by a structural variant N57D of the first epidermal growth factor domain. AB - We have previously described a kindred with factor VII (FVII) deficiency whose members exhibited reduced procoagulant activity relative to FVII antigen concentration. In this report, the molecular genetic basis of the FVII defect has been determined to be a heterozygous substitution of Asp for Asn at position 57 in the first epidermal growth factor (EGF) domain. Recombinant FVII (N57D) cDNA was created by site-directed mutagenesis and transiently expressed in human 293 cells. The transfected cells synthesized an immunoprecipitable protein with an apparent molecular weight of 50 kD. Quantitation of expression by FVII enzyme linked immunosorbent assay indicated that mutant protein yields were consistently low, typically 10% to 30% of wild-type FVII. FVII (N57D) protein did not accumulate intracellularly, and Northern blot analysis indicated equivalent FVII mRNA levels in 293 cells expressing either wild-type FVII or FVII (N57D). Secreted FVII (N57D) protein did not bind tissue factor, exhibited no procoagulant activity, and failed to bind a conformation-dependent monoclonal antibody specific for the first EGF domain of FVII. Molecular modeling of the first EGF domain of FVII predicted that the N57D amino acid substitution would disrupt tertiary bonding structure. We conclude that the N57D mutation affects folding of the first EGF domain of FVII resulting in decreased cellular secretion of a mutant FVII molecule, which is unable to bind tissue factor and is therefore biologically inactive. PMID- 9414279 TI - Identification of a large deletion, spanning exons 4 to 11 of the human factor XIIIA gene, in a factor XIII-deficient family. AB - Inherited deficiency of factor XIIIA subunit (FXIIIA) is an autosomal recessive disorder that is characterized by a life-long bleeding tendency and complications in wound healing. Molecular genetic studies have shown the deficiency can be due to small sequence changes within the FXIIIA gene, such as point mutations or microdeletions. On molecular analysis of the FXIIIA gene in an FXIII-deficient patient, of United Kingdom origin, we identified a putative homozygous missense mutation, Arg408Gln. However, the father of this patient is homozygous normal for arginine at codon 408. Having proved paternity in this pedigree by microsatellite analysis, we examined the FXIIIA RNA of the patient by reverse transcriptase polymerase chain reaction and found the paternal allele to lack exons 4 through 11 inclusive. Hence, a huge deletion extending from intron 3 to intron 11 and the Arg408Gln mutation are jointly responsible for FXIIIA deficiency in this family. This is the first finding of such a large deletion in the FXIIIA gene. PMID- 9414280 TI - A novel P-selectin glycoprotein ligand-1 monoclonal antibody recognizes an epitope within the tyrosine sulfate motif of human PSGL-1 and blocks recognition of both P- and L-selectin. AB - Interactions between P-selectin and P-selectin glycoprotein ligand-1 (PSGL-1) mediate the earliest "rolling" of leukocytes on the lumenal surface of endothelial cells at sites of inflammation. Previously, PSGL-1 has been shown to be the primary mediator of interactions between neutrophils and P-selectin, but studies on the ability of PSGL-1 to mediate interactions between P-selectin and other subsets of leukocytes have yielded variable and conflicting results. A novel IgG monoclonal antibody (MoAb) to human PSGL-1 was generated, and the specificity of this MoAb was confirmed by both flow cytometric analysis and Western blotting of cells transfected with human PSGL-1. This newly developed MoAb, KPL1, inhibited interactions between P-selectin expressing COS cells and either HL60 cells, neutrophils, or lymphocytes. Furthermore, KPL1 completely inhibited interactions between P-selectin and either purified CD4 T cells or neutrophils in a flow assay under physiological conditions, but had no effect on interactions of T cells or neutrophils with E-selectin. In addition, KPL1 blocked interactions between lymphoid cells transfected with L-selectin and COS cells expressing PSGL-1. The KPL1 epitope was mapped to a site within a consensus tyrosine sulfation motif of PSGL-1, previously shown to be essential for interaction with P-selectin and now shown to be essential for interaction with L selectin, and to be distinct from the epitope identified by the PL1 function blocking anti-PSGL-1 MoAb. Two-color flow cytometry of normal leukocytes showed that while natural killer (NK) cells (CD16(+)), monocytes, CD4 and CD8 T cells, and alpha/beta and gamma/delta T cells were uniformly positive for PSGL-1, B cells expressed low levels of the KPL1 epitope. This low level of KPL1 staining was also observed immunohistologically in germinal centers, which had no detectable KPL1 staining, whereas T-cell areas (interfollicular region) were positive for KPL1. Interestingly, plasma cells in situ and interleukin-6 dependent myeloma cell lines were KPL1(+). Thus, PSGL-1 is expressed on essentially all blood neutrophils, NK cells, B cells, T cells, and monocytes. Variation in tyrosine sulfation during B-cell differentiation may affect the ability of B cells to interact with P- and L-selectin. PMID- 9414281 TI - Further analysis of interleukin-2 receptor subunit expression on the different human peripheral blood mononuclear cell subsets. AB - We have investigated the expression of the three components of the interleukin-2 receptor (IL-2Ralpha, IL-2Rbeta, and IL-2Rgamma) on the surface of the various peripheral blood mononuclear cell (PBMC) subsets by flow cytometry analysis. The PBMC were immediately isolated (ficoll) from blood collected on heparin as anticoagulant. The three IL-2R components are absent or only marginally detectable on CD4 T lymphocytes. No expression of the IL-2R chains is found for the B lymphocytes. In most donors, the three chains are not detectable on CD8 T lymphocytes, but for a few of them, IL-2Rbeta or IL-2Rgamma are clearly expressed. CD56 high (IL-2Ralpha+) and CD56 low (IL-2Ralpha-) natural killer (NK) cells express IL-2Rbeta, but not IL-2Rgamma. IL-2Rgamma is expressed by monocytes of all donors although with variable intensity. When blood is collected on other anticoagulants or when cells are isolated 1 day after collection, IL-2Ralpha, IL 2Rbeta, and IL-2Rgamma are largely expressed on the surface of most PBMC. This observation provides a possible explanation for divergent data previously reported on IL-2R expression. Finally, we show that IL-2Rgamma, which is not detectable on the cell surface of lymphocytes, is nevertheless expressed and stored as an intracellular component. This result is in agreement with the constitutive expression of the IL-2Rgamma gene and suggests a specific regulatory mechanism for IL-2Rgamma membrane translocation. PMID- 9414282 TI - Generation of plasma cells from peripheral blood memory B cells: synergistic effect of interleukin-10 and CD27/CD70 interaction. AB - B cells can differentiate into the antibody-secreting cells, plasma cells, whereas the crucial signals that positively control the entry into the pathway to plasma cells have been unclear. Triggering via CD27 by CD27 ligand (CD70) on purified peripheral blood B cells yielded an increase in the number of plasma cells in the presence of interleukin-10 (IL-10). Differentiation into plasma cells by a combination of IL-10 and CD70 transfectants occurred in CD27+ B cells but not in CD27- B cells. Moreover, addition of IL-2 to the IL-10 and CD70 transfect activation system greatly induced differentiation into plasma cells. In the presence of only IL-2, IL-4, or IL-6, CD70 transfectants did not promote differentiation into plasma cells. On the other hand, CD40 signaling increased the expansion of a B-cell pool from peripheral blood B cells primarily activated by IL-2, IL-10, and anti-CD40 monoclonal antibody (MoAb). Finally, CD27 signaling also rescued B cells from IL-10-mediated apoptosis. These data demonstrate that CD27 ligand (CD70) is a key molecule to prevent the IL-10-mediated promotion of apoptosis and to direct the differentiation of CD27+ memory B cells toward plasma cells in cooperation with IL-10. PMID- 9414283 TI - Diagnosis and clinical course of autoimmune neutropenia in infancy: analysis of 240 cases. AB - Primary autoimmune neutropenia (AIN) is caused by granulocyte-specific autoantibodies and occurs predominantly in infancy. Clinical presentation and diagnosis have not been well established, resulting in burdening diagnostic investigations and unnecessary treatment with granulocyte colony-stimulating factor (G-CSF). In the present study, clinical, laboratory, and immunologic data of 240 infants with primary AIN were evaluated. Suspected association with parvovirus B19 infection was investigated using serologic and DNA-based methods. Primary AIN was mainly diagnosed at the age of 5 to 15 months but was observed as early as day 33 of life. In 90% of the cases, AIN was associated with benign infections despite severe neutropenia. Spontaneous remission, shown by 95% of the patients, usually occurred within 7 to 24 months. Autoantibodies in the patient's sera were not always present, and screening had to be repeated several times until antibody detection succeeded. About 35% of the autoantibodies showed preferential binding to granulocytes from NA1 and NA2 homozygous donors. Bone marrow was typically normocellular or hypercellular, with a variably diminished number of segmented granulocytes. A significant association with parvovirus B19 infection was not found. Symptomatic treatment with antibiotics was sufficient in most patients. Eighty-nine percent of the patients received antibiotics (cotrimoxazole) for prophylaxis of infections. For severe infections or for surgical preparation, G-CSF, corticosteroids, and intravenous IgG were administered, resulting in increased neutrophil counts in 100%, 75%, and 50% of the patients treated, respectively. In combination with the detection of granulocyte-specific antibodies, the typical clinical picture allowed diagnosis of AIN without burdening investigations. Treatment with G-CSF was found to be a reliable alternative to temporarily increase the neutrophil count. PMID- 9414284 TI - Interleukin-4 promotes the development of tryptase and chymase double-positive human mast cells accompanied by cell maturation. AB - Human cultured mast cells (HCMCs) grown from cord blood mononuclear cells in the presence of stem cell factor (SCF) and interleukin-6 (IL-6) expressed tryptase but no or low chymase in their cytoplasm. The addition of IL-4 to these cells strikingly increased chymase expression. Consequently, the activity of chymase was significantly higher in IL-4-treated mast cells than that in IL-4-nontreated mast cells, whereas the activity of tryptase and histamine content were comparable in both cells. Electron microscopic immunocytochemistry also showed that secretary granules containing chymase increased in IL-4-treated mast cells. Interestingly, the IL-4-induced increase of chymase expression in HCMCs was accompanied by morphological maturation of the cells. Cytoplasmic projections were few in IL-4-nontreated HCMCs, and a small number of secretary granules were observed, most of which were empty or partially filled with discrete scrolls with rough particles showing immaturity. In contrast, IL-4-treated HCMCs had extremely abundant cytoplasmic projections and had many secretary granules filled with electron-dense crystal materials. Taken together, immature HCMCs grown only with SCF and IL-6 expressed tryptase with no or a low amount of chymase, and addition of IL-4 promoted cell maturation together with the expression of both tryptase and a high amount of chymase. Our findings will raise a possibility of a linear pathway of human mast cell development from tryptase single positive mast cells into tryptase and chymase double positive mast cells as the cells mature and will suggest that this maturation process is promoted by IL-4. PMID- 9414285 TI - Interleukin-12-activated natural killer cells recognize B7 costimulatory molecules on tumor cells and autologous dendritic cells. AB - Activation of natural killer (NK) cells in the presence of interleukin-12 (IL-12) augments the capacity of these effector cells to recognize B7-1- and B7-2 expressing target cells. These effector cells also efficiently lyse autologous B7 positive progenitor or organ-derived dendritic cells, suggesting a physiologic regulatory pathway between IL-12, NK cells, and B7-expressing antigen-presenting cells. Although IL-12-activated NK cells secreted higher levels of interferon gamma, this cytokine did not play a role in synergistic effects of IL-12 and B7 on NK activation. The B7-counterreceptor was found to be selectively upregulated on IL-2/IL-12 as compared with IL-2-activated NK cells. CD28 is functionally involved in the recognition of B7 on target cells since IL-2/IL-12-activated NK cells derived from CD28 knockout mice were strongly reduced in their capacity to lyse syngeneic B7-positive tumor cells as well as antigen-presenting cells. However, recognition of B7 on allogeneic targets did not require the expression of CD28 on the IL-2/IL-12-activated NK cells. Hence, IL-12 triggers the expression of both CD28-dependent and CD28-independent mechanisms that allow NK cells to eliminate B7-positive target cells including autologous dendritic cells. PMID- 9414286 TI - Differential cytotoxicity of cord blood and bone marrow-derived natural killer cells. AB - Allogeneic cord blood is now being widely used as a source of stem cells for hematologic reconstitution after myeloablative therapy, with reported significantly lower levels of graft-versus-host disease (GVHD) compared with the use of allogeneic bone marrow (BM). This study was undertaken to investigate biologic aspects of natural killer (NK) cell activity, as recognized effector cells of the GVHD and graft-versus-leukemia (GVL) response, from cord blood and conventional BM. NK-cell activity levels of freshly isolated cells from cord blood and BM against K562 targets were comparable. Lymphokine activated killer (LAK) cells from both hematopoietic cell sources were compared for their ability to kill target cells by necrotic or apoptotic mechanisms using specific target cell lines. Cord blood cells had significantly higher necrosis-mediated cytotoxic activity against Daudi target cells compared with BM-derived cells. Cord blood LAK cells had relatively high levels of apoptotic-mediated cytotoxicity against YAC-1 target cells, whereas BM-derived LAK cells were unable to induce apoptosis in these cells. Interleukin-2 (IL-2) induced significant granzyme B activity in cord cells in contrast to BM cells, in which very little activity was measured. Western blotting confirmed these findings, with IL-2 inducing granzyme B protein expression in cord cells but not detectable levels in BM cells. BM cells had significantly lower cell surface expression of IL-2R and prolonged culture in IL 2 was only partially able to restore their deficient apoptotic cytotoxic activity. Thus, major differences exist between cord blood-derived and BM-derived mononuclear cells with respect to their NK-cell-associated cytotoxic behavior. This could have important implications for stem cell transplantation phenomena, because it suggests that cord blood may have increased potential for a GVL effect. PMID- 9414287 TI - Transcriptional regulatory elements within the first intron of Bruton's tyrosine kinase. AB - Defects in the gene for Bruton's tyrosine kinase (Btk) result in the disorder X linked agammaglobulinemia (XLA). Whereas XLA is characterized by a profound defect in B-cell development, Btk is expressed in both the B lymphocyte and myeloid cell lineages. We evaluated a patient with XLA who had reduced amounts of Btk transcript but no abnormalities in his coding sequence. A single base-pair substitution in the first intron of Btk was identified in this patient, suggesting that this region may contain regulatory elements. Using reporter constructs we identified two transcriptional control elements in the first 500 bp of intron 1. A strong positive regulator, active in both pre-B cells and B cells, was identified within the first 43 bp of the intron. Gel-shift assays identified two Sp1 binding sites within this element. The patient's mutation results in an altered binding specificity of the proximal Sp1 binding site. A negative regulator, active in pre-B cells only, was located between base pairs 281 and 491 of the intron. These findings indicate that regulation of Btk transcription is complex and may involve several transcriptional regulatory factors at the different stages of B-cell differentiation. PMID- 9414288 TI - Gene immunotherapy in murine acute myeloid leukemia: granulocyte-macrophage colony-stimulating factor tumor cell vaccines elicit more potent antitumor immunity compared with B7 family and other cytokine vaccines. AB - In an attempt to explore novel treatment modalities in acute myeloid leukemia (AML), we studied the role of costimulatory and cytokine gene immunotherapy in murine AML. We have previously shown that leukemic mice can be cured with CD80 transfected leukemic cells (B7. 1-AML vaccine) administered early in the course of the disease and that the failure B7.1-AML vaccines administered late cannot be attributed to immunosuppression induced by tumor growth. CD8+ T cells, which are necessary for tumor rejection, are activated rather than suppressed during the first half of the leukemic course in nonvaccinated mice. In this report, we question whether CD86 (B7.2) or the cytokines granulocyte-macrophage colony stimulating factor (GM-CSF), interleukin-4 (IL-4), or tumor necrosis factor-alpha (TNF-alpha) can improve the vaccination potential of AML cells. The choice of cytokines was based on their combined and alone as well ability to direct the differentiation of CD34+ cells into potent antigen-presenting dendritic cells in vitro. Our studies show that (1) mice vaccinated with a leukemogenic number of AML cells engineered to express B7.2 (B7.2-AML) or to secrete GM-CSF, IL-4, or TNF-alpha (GM-, IL-4-, TNF-alpha-AML) do not develop leukemia; (2) GM-AML cells are tumorigenic in sublethally irradiated SJL/J mice but not in Swiss nu/nu mice, indicating that killing of tumor cells is not T-cell-dependent; (3) vaccines with irradiated GM-AML, but not B7.2-, IL-4-, or TNF-alpha-AML cells, can elicit leukemia-specific protective and therapeutic immunity; and (4) in head-to-head comparison experiments, vaccination with irradiated GM-AML is more potent than B7.1-AML, curing 80% and providing 20% prolonged survival of the leukemic mice at week 2, as opposed to cures only up to 1 week with B7.1-AML vaccines. These preclinical data emphasize that GM-CSF gene immunotherapy deserves clinical evaluation in AML. PMID- 9414289 TI - Molecular delineation of 13q deletion boundaries in 20 patients with myeloid malignancies. AB - Fluorescent in situ hybridization (FISH) analysis with a panel of DNA probes for 13q13.1-q14.3 was performed on 20 cases of myeloid malignancies, of which 17 showed a del(13)(q) and three had translocations affecting 13q. By chromosome morphology, deletions consistently involved bands q14 and q21. In addition to confirming the chromosome data, FISH allowed us to delineate a commonly deleted region that was flanked by YAC 833A2 and YAC 854D4. Three cases with 13q translocations unexpectedly showed accompanying cryptic microdeletions of 13q, and in one case the commonly deleted region could be narrowed to a genomic segment, which includes YAC 937C7, RB1, and YAC 745E3. Homozygous deletions were not detected. This region overlaps with the smallest deleted region of 13q14 in chronic lymphocytic leukemia. PMID- 9414290 TI - VH gene analysis of clonally related IgM and IgG from human lymphoplasmacytoid B cell tumors with chronic lymphocytic leukemia features and high serum monoclonal IgG. AB - An unusual group of human B-cell tumors with cellular features of chronic lymphocytic leukemia or lymphoplasmacytoid leukemia, together with high levels of a monoclonal IgG serum protein, has been investigated. Analysis of tumor-derived VH genes of neoplastic B lymphocytes was used to determine the clonal relationship between the IgM expressed or secreted by the tumor cells and the IgG serum paraprotein. In all five cases, VH gene sequences showed transcripts of IgM and IgG of common clonal origin. Sequences were derived from VH3 (4 of 5) and VH1 (1 of 5) families and were all highly somatically mutated with strong evidence for antigen selection. There was no intraclonal variation detectable in either IgM or IgG sequences. In 3 of 5 cases, in which monoclonal IgM and IgG were found in serum, the VH genes combined to Cmu or Cgamma showed identical mutational patterns. However, in 2 of 5 cases, in which IgM was confined to cell expression with only monoclonal IgG in serum, sequences of the VH transcripts of IgM and IgG showed many shared mutations but also numerous differences. In these cases, the level of mutation was similar in IgM and IgG and both appeared to be antigen selected. In summary, the final neoplastic event in this group of tumors has apparently occurred at the point of isotype switch from IgM to IgG, leading to dual isotype synthesis. In the group that secreted both isotypes, the mutation pattern was identical, indicating either synthesis by a single cell, or silencing of mutational activity before switching. In the group that did not secrete IgM, cells of each isotype were distinct and reflected a divergent mutational history. PMID- 9414291 TI - Unbalanced expression of bcl-2 family proteins in follicular lymphoma: contribution of CD40 signaling in promoting survival. AB - Although highly responsive, advanced stage follicular lymphoma (FL) is not curable with conventional treatment. This relative resistance is thought to be due to the t(14;18) that results in the constitutive overexpression of the death inhibiting protein bcl-2. However, the observation that FL cells are sensitive to treatment in vivo and prone to apoptosis on in vitro culture questions whether bcl-2 alone is responsible for the pathogenesis and clinical behavior of this disease. Therefore, multiple genes are likely to be involved in both the lymphomagenesis and the clinical course of FL. We examined whether expression of other bcl-2 family genes might also be operative. Here, we show that FL cells display a different pattern of expression of bcl-2 family proteins from normal germinal center (GC) B cells that are thought to be their normal counterpart. FL cells express the death-suppressor proteins bcl-2, bcl-xL, and mcl-1; whereas GC B cells express bcl-xL and mcl-1 but also the proapoptotic proteins bax-alpha and bad. Although maintaining constitutive levels of bcl-2 and mcl-1, FL cells are not protected from apoptosis when cultured in vitro. Their propensity to undergo apoptosis is temporally associated with downregulation of bcl-xL. More importantly, activation of FL cells via CD40 not only prevents downregulation but increases the level of bcl-xL expression and results in promotion of survival. These results support the hypothesis that the overexpression of bcl-2 is not the only antiapoptotic mechanism responsible for the pathogenesis of FL. Survival of FL cells is determined by a number of death-inhibiting proteins, among which bcl xL appears to have the most critical role. Moreover, these findings are consistent with the hypothesis that, although FL cells are malignant, they respond to microenvironmental signals such as CD40L that appear to contribute to their survival through the upregulation of death-inhibiting proteins. PMID- 9414292 TI - Somatic triple mosaicism in a carrier of X-linked chronic granulomatous disease. AB - The X-linked form of chronic granulomatous disease (CGD) is caused by mutations in the CYBB gene, which encodes the 91-kD subunit of the flavocytochrome b558, a component of the superoxide-generating nicotinamide adenine dinucleotide phosphate (NADPH) oxidase in phagocytic leukocytes. Mutations in this gene are very heterogeneous and often unique for one family. Here we report on a family with two patients (brothers), one with a 3-kb deletion comprising exon 5 and the other with a 3.5-kb deletion comprising exons 6 and 7 of the CYBB gene. Sequence analysis of polymerase chain reaction (PCR)-amplified genomic DNA proved these deletions to be overlapping for 35 bp. Analysis by restriction fragment length polymorphism of genomic DNA from the mother's leukocytes showed her to be a carrier of both deletions in addition to the normal CYBB sequence. This triple somatic mosaicism was confirmed with PCR-amplified genomic and complementary DNA. The presence of the normal CYBB gene in the mother was also proven by the finding of normal superoxide-generating neutrophils in addition to cells lacking this ability. Triple X syndrome was excluded. These findings suggest that the mutations are the result of an event in early embryogenesis of the mother, possibly involving a mechanism like sister chromatid exchange. PMID- 9414293 TI - Interleukin-6 induces monocyte chemotactic protein-1 in peripheral blood mononuclear cells and in the U937 cell line. AB - Induction of chemokine gene expression from peripheral blood mononuclear cells (PBMCs) stimulated by proinflammatory cytokines plays an important role in both wound repair and response to infectious agents. In the present study, we show that the proinflammatory cytokine interleukin-6 (IL-6) potently induced mRNA expression and secretion of the CC chemokine monocyte chemotactic protein 1 (MCP 1) in PBMCs. In addition, because human immunodeficiency virus (HIV) infection in vivo and in vitro has been shown to dysregulate the production of and/or the response to cytokines, PBMCs from both healthy uninfected and HIV-infected individuals were studied for their constitutive and IL-6-induced expression of MCP-1. No substantial differences were observed between the two groups of individuals. In addition, IL-6 upregulated MCP-1 expression in the promonocytic cell line U937 and in its chronically HIV-infected counterpart, U1. In these cell lines, IL-6 selectively induced MCP-1 and not other chemokines, including regulated upon activation normal T cells expressed and secreted (RANTES), macrophage inflammatory protein-1alpha (MIP-1alpha), MIP-1beta, and IL-8. IL-6 induction of MCP-1 was partially inhibited by hydrocortisone in U1 cells. Thus, IL-6 activates PBMCs to secrete MCP-1, a CC chemokine pivotal for monocyte recruitment in tissue and organs in which important inflammatory events occur. PMID- 9414294 TI - Blood polymorphonuclear leukocytes from the majority of sickle cell patients in the crisis phase of the disease show enhanced adhesion to vascular endothelium and increased expression of CD64. AB - There is increasing interest in the role of blood polymorphonuclear leukocytes (PMNs) in the pathogenesis of sickle cell crisis. We studied the adherence of PMNs from 18 sickle cell patients in crisis, 25 out of crisis, and 43 healthy subjects (controls) to monolayers of human umbilical cord endothelium that were either untreated or pretreated with tumor necrosis factor alpha (TNFalpha). Overall, the PMNs from patients in crisis were more adherent than control PMNs to untreated endothelial monolayers (mean 53% increase; P < .001) and TNFalpha treated monolayers (mean 41% increase; P < .002). Increased adhesiveness was not associated with an abnormal expression of CD11a, CD11b, CD11c, CD18, CD62L, or CD15. There was an increase in the number of PMNs expressing CD64 in patients in crisis (median value, 44%) compared with patients out of crisis (median, 21%; P = .025) and controls (median, 6.5%; P < .001). Sera from patients in crisis had normal levels of granulocyte colony-stimulating factor, granulocyte-macrophage colony-stimulating factor, interferon-gamma, TNFalpha, interleukin-1 (IL-1), IL 6, or IL-8 and did not modify the adherence of PMNs or their expression of CD64. Only IFN-gamma induced CD64 expression on PMNs, but this effect was not associated with enhanced binding to endothelium. Because PMNs bound to endothelial monolayers were CD64(+) and CD64-enriched PMNs were 7 times more adherent to endothelial monolayers than CD64-depleted PMNs, it is likely that CD64 is a marker of adherent PMNs. Two of the three anti-CD64 antibodies used in our antibody blocking studies (clones 32.2 and 197) partially inhibited the binding of sickle cell PMNs to untreated endothelium (mean inhibitions of 33% [P = .01] and 21% [P = .03], respectively), whereas only one (clone 197) inhibited binding to TNFalpha-treated endothelium (mean inhibition, 29%; P = . 004). In some patients with sickle cell disease, an enhanced PMN adhesion to vascular endothelium could contribute to the vascular occlusion that characterizes the acute crisis of the disease. PMID- 9414295 TI - DNA cross-linker-induced G2/M arrest in group C Fanconi anemia lymphoblasts reflects normal checkpoint function. AB - Cells from individuals with Fanconi anemia (FA) arrest excessively in the G2/M cell cycle compartment after exposure to low doses of DNA cross-linking agents. The relationship of this abnormality to the fundamental genetic defect in such cells is unknown, but many investigators have speculated that the various FA genes directly regulate cell cycle checkpoints. We tested the hypothesis that the protein encoded by the FA group C complementing gene (FAC) functions to control a cell cycle checkpoint and that cells from group C patients (FA[C]) have abnormalities of cell cycle regulation directly related to the genetic mutation. We found that retroviral transduction of FA(C) lymphoblasts with wild-type FAC cDNA resulted in normalization of the cell cycle response to low-dose mitomycin C (MMC). However, when DNA damage was quantified in terms of cytogenetic damage or cellular cytotoxicity, we found similar degrees of G2/M arrest in response to equitoxic amounts of MMC in FA(C) cells as well as in normal lymphoblasts. Similar results were obtained using isogenic pairs of uncorrected, FAC- or mock corrected (neo only) FA(C) cell lines. To test the function of other checkpoints we examined the effects of hydroxyurea (HU) and ionizing radiation on cell cycle kinetics of FA(C) and normal lymphoblasts as well as with isogenic pairs of uncorrected, FAC-corrected, or mock-corrected FA(C) cell lines. In all cases the cell cycle response of FA(C) and normal lymphoblasts to these two agents were identical. Based on these studies we conclude that the aberrant G2/M arrest that typifies the response of FA(C) cells to low doses of cross-linking agents does not represent an abnormal cell cycle response but instead represents a normal cellular response to the excessive DNA damage that results in FA(C) cells following exposure to low doses of cross-linking agents. PMID- 9414296 TI - Cerebrovascular accidents in sickle cell disease: rates and risk factors. AB - Cerebrovascular accident (CVA) is a major complication of sickle cell disease. The incidence and mortality of and risk factors for CVA in sickle cell disease patients in the United States have been reported only in small patient samples. The Cooperative Study of Sickle Cell Disease collected clinical data on 4,082 sickle cell disease patients enrolled from 1978 to 1988. Patients were followed for an average of 5.2 +/- 2.0 years. Age-specific prevalence and incidence rates of CVA in patients with the common genotypes of sickle cell disease were determined, and the effects of hematologic and clinical events on the risk of CVA were analyzed. The highest rates of prevalence of CVA (4.01%) and incidence (0.61 per 100 patient-years) were in sickle cell anemia (SS) patients, but CVA occurred in all common genotypes. The incidence of infarctive CVA was lowest in SS patients 20 to 29 years of age and higher in children and older patients. Conversely, the incidence of hemorrhagic stroke in SS patients was highest among patients aged 20 to 29 years. Across all ages the mortality rate was 26% in the 2 weeks after hemorrhagic stroke. No deaths occurred after infarctive stroke. Risk factors for infarctive stroke included prior transient ischemic attack, low steady-state hemoglobin concentration and rate of and recent episode of acute chest syndrome, and elevated systolic blood pressure. Hemorrhagic stroke was associated with low steady-state hemoglobin and high leukocyte count. PMID- 9414297 TI - Long-term trial of deferiprone in 51 transfusion-dependent iron overloaded patients. AB - Fifty-one transfusion-dependent iron-loaded adult patients (38 with thalassemia major) were treated with the orally active iron chelator deferiprone (1,2 dimethyl-3-hydroxypyrid-4-one, L1) at a dose of 75 mg/kg/d (range, 50 to 79). Twenty patients discontinued the drug and five died after a mean of 18.7 months (range, 4 to 35). Of the 20, 5 had arthropathy, 5 had gastrointestinal symptoms, 4 had a rising serum ferritin, 3 had agranulocytosis or neutropenia, 1 had tachycardia, 1 had renal failure, and 1 went abroad. Twenty-six patients continued deferiprone for a mean of 39.4 months (range, 12 to 49). Among these patients, there was no overall significant change in serum ferritin (initial mean, 2,937 microg/L; range, 980 to 5,970; final mean, 2,323 microg/L; range, 825 to 5,970) or in urine iron excretion (initial mean, 31.2 mg/24 h; range, 16.3 to 58. 2; final mean, 32.1 mg/24 h; range, 9.4 to 75.8), implying no overall change in iron stores. When the patients who had received deferiprone for longer than 3 years were considered separately, there was also no significant change in serum ferritin or urinary iron excretion. The initial serum ferritin levels in the 26 patients who continued deferiprone treatment were significantly lower than in those who discontinued the drug (P < .01). The liver iron content in 17 patients who had received deferiprone for 24 to 48 months ranged from 5.9 to 41.2 mg/g dry weight, 50% having levels above 15.0 mg, a level associated with a high risk of cardiac disease due to iron overload. In this study the drug caused fewer side effects and was more effective at maintaining iron status among patients previously well chelated and with lower initial serum ferritin levels. PMID- 9414298 TI - Epidemiology of anemia in human immunodeficiency virus (HIV)-infected persons: results from the multistate adult and adolescent spectrum of HIV disease surveillance project. AB - To study the incidence of, the factors associated with, and the effect on survival of anemia in human immunodeficiency virus (HIV)-infected persons, we analyzed data from the longitudinal medical record reviews of 32,867 HIV-infected persons who received medical care from January 1990 through August 1996 in clinics, hospitals, and private medical practices in nine United States cities. We calculated the 1-year incidence of anemia (a hemoglobin level of <10 g/dL or a physician diagnosis of anemia); the adjusted odds ratios showing excess risk of anemia associated with demographic factors, prescribed therapies, and concurrent diseases; the risk of death for patients who developed anemia compared with risk for patients who did not develop anemia; and, of patients who did develop anemia, the risk of death for those who did not recover from anemia compared with the risk for those who did recover. The 1-year incidence of anemia was 36.9% for persons with one or more acquired immunodeficiency syndrome (AIDS)-defining opportunistic illnesses (clinical AIDS), 12.1% for patients with a CD4 count of less than 200 cells/micron or CD4 percentage of <14 but not clinical AIDS (immunologic AIDS), and 3.2% for persons without clinical or immunologic AIDS. Of anemia diagnoses, 22% were identified by physicians as drug related. Incidence of anemia was associated with clinical AIDS, immunologic AIDS, neutropenia, thrombocytopenia, bacterial septicemia, black race, female sex, prescription of zidovudine, fluconazole, and ganciclovir, and lack of prescription of trimethoprim-sulfamethoxazole. The increased risk of death associated with anemia differed by first CD4 count: for patients with a CD4 count of >/=200 cells/microL at the beginning of the survival analysis, the risk of death was 148% (99% confidence interval [CI], 114 to 188) greater for those who developed anemia; for patients whose first CD4 count was <200 cells/microL, the risk of death was 56% (99% CI, 43 to 71) greater for those in whom anemia developed. For persons in whom anemia developed, the risk of death was 170% (99% CI, 132 to 203) greater for persons who did not recover from anemia compared with those who did recover. Anemia is a frequent complication of HIV infection, and its incidence is associated with progression of HIV disease, prescription of certain chemotherapeutics, black race, and female sex. Anemia, particularly anemia that does not resolve, is associated with shorter survival of HIV-infected patients. PMID- 9414299 TI - Gamma-globin gene promoter elements required for interaction with globin enhancers. AB - Normal expression of the human beta-globin domain genes is dependent on at least three types of regulatory elements located within the beta-globin domain: the locus control region (LCR), globin enhancer elements (3'beta and 3'Agamma), and the individual globin gene promoter and upstream regions. It has been postulated that regulation occurs through physical interactions between factors bound to these elements, which are located at considerable distances from each other. To identify the elements required for promoter-enhancer interactions from a distance, we have investigated the expression of the wild-type, truncated, and mutated gamma-globin promoters linked to the 5'HS2 enhancer. We show that in K562 cells, 5'HS2 increases activity approximately 20-fold from both a wild-type and truncated (-135 --> +25) gamma promoter and that truncation or site-directed mutagenesis of the tandem CCAAT boxes eliminated the enhancement by 5'HS2. Mutation of the gamma-globin gene promoter GATA-1 binding sites did not decrease either promoter strength or enhancement of activity by 5'HS2. To determine if enhanced expression of gamma-globin gene promoters carrying mutations associated with hereditary persistence of fetal hemoglobin (HPFH) was due to greater interactions with enhancers, we linked these HPFH gamma-globin gene promoters to 5'HS2 and demonstrated a twofold to threefold higher expression than the corresponding wild-type promoter plus enhancer in MEL cells. Addition of the Agamma-globin gene 3' enhancer to a plasmid containing the gamma-globin gene promoter and 5'HS2 did not further enhance promoter strength. Furthermore, we have demonstrated that the previously identified core 5'HS2 enhancer (46-bp tandem AP-1/NF-E2 sites) increased expression only when located 5', but not 3', to the gamma-globin-luciferase reporter gene, suggesting that its enhancer effect is not by DNA looping. Our results suggest that CCAAT boxes, but not GATA or CACCC binding sites, are required for interaction between the gamma-globin promoter and the LCR/5'HS2 and that regulatory elements in addition to the core enhancer may be required for the enhancer to act from a distance. PMID- 9414300 TI - A point mutation in the bulge of the iron-responsive element of the L ferritin gene in two families with the hereditary hyperferritinemia-cataract syndrome. AB - The molecular basis for the recently described hereditary hyperferritinemia cataract syndrome is the presence of a mutation in the iron-responsive element (IRE) of the L ferritin gene, located on chromosome 19q13.3-13.4. Two mutations have been reported so far, altering adjacent nucleotides in the IRE loop, in a region that has been extensively studied in vitro and shown to mediate high affinity interaction with the iron-responsive protein. In this report, we describe two families with a new mutation in the bulge of the IRE stem, and we show that this mutation alters the protein-binding affinity of the IRE in vitro to the same extent as the loop mutation. In addition, we present evidence that some variability in the age of onset of cataract can be associated with this genetic syndrome, probably because of additional genetic or environmental factors that modulate the penetrance of the L ferritin defect in the lens. We confirm that the patients do not have increased iron stores despite the persistence of elevated serum ferritin levels and that, accordingly, they do not tolerate well venesection therapy. Further studies will be necessary to elucidate the mechanism responsible for the onset of cataract. PMID- 9414301 TI - Mechanisms of long-term donor-specific allograft survival induced by pretransplant infusion of lymphocytes. AB - Pretransplantation donor-specific transfusion (DST) can enhance allograft survival in man and animals. However, due to the lack of a specific marker to identify donor-reactive cells in vivo in man and normal (nontransgenic) animals, the underlying mechanism remains unknown. In this study, we use 2CF1 transgenic mice expressing a transgenic T-cell receptor (TCR) specifically recognizing Ld, a major histocompatibility complex (MHC) class I molecule, to delineate the role of DST in long-term skin allograft survival and its underlying mechanisms. Our main findings include: (1) in the absence of any other immunosuppressive treatment, a single dose pretransplantation infusion of viable splenocytes from an Ld+ donor is sufficient to induce permanent donor-specific skin allograft survival in 2CF1 anti-Ld TCR transgenic mice; (2) DST leads to a deletion of the majority (>60%) of donor-reactive T cells in the periphery of the recipient. However, deletion does not necessarily result in tolerance; (3) remaining donor-reactive T cells from DST-treated mice are fully responsive to Ld in vitro, and can suppress the antidonor response of naive T cells in vitro only when exogenous interleukin (IL) 4 is provided; and (4) the sera level of IL-4 in DST-treated tolerant mice is significantly increased. These results suggest that the generation of a subset of T cells with the potential to specifically inhibit antidonor responses, together with promotion of IL-4 production in recipients, may be important mechanisms for the induction and maintenance of antigen-specific tolerance. PMID- 9414303 TI - The umbilical cord blood alphabeta T-cell repertoire: characteristics of a polyclonal and naive but completely formed repertoire. AB - Umbilical cord blood (CB) constitutes a promising alternative to bone marrow for allogeneic transplantation and is increasingly used because of the reduced severity of graft-versus-host disease after CB transplantation. We have compared the T-cell receptor beta chain (TCRB) diversity of CB lymphocytes with that of adult lymphocytes by analyzing the complementarity determining region 3 (CDR3) size heterogeneity. In marked contrast to adult samples, we observed bell-shaped profiles in all of the 22 functional beta-chain variable (BV) subfamilies that reflect the lack of prior antigenic stimulation in CB samples. However, the mean CDR3 size and BV usage were comparable between CB and adult samples. BJ2 (65%) segments were used preferentially to BJ1 (35%), especially BJ2S7, BJ2S5, BJ2S3, and BJ2S1, in both CB and in adult lymphocytes. We therefore conclude that although naive as reflected by the heterogeneity of the CDR3 size, the TCRBV repertoire appears fully constituted at birth. The ability to expand TCRB subfamilies was confirmed by stimulation with staphylococcal superantigens toxic shock syndrome toxin-1 and staphylococcal enterotoxin A. This study provides the basis for future analysis of the T-cell repertoire reconstitution following umbilical CB transplantation. PMID- 9414302 TI - Lymphoma cell burden in progenitor cell grafts measured by competitive polymerase chain reaction: less than one log difference between bone marrow and peripheral blood sources. AB - A controversy persists in autologous transplantation as to which source of progenitor cells, bone marrow (BM) or peripheral blood (PB), contains the lowest number of contaminating lymphoma cells, and how mobilization procedures affect these numbers. To accurately measure the number of non-Hodgkin's lymphoma (NHL) cells harboring the bcl-2/immunoglobulin H (IgH) rearrangement in progenitor cell grafts, we developed a nested quantitative competitive polymerase chain reaction assay (QC-PCR). DNA from lymph nodes of four patients with NHL were cloned into the pSK(+) vectors to generate four internal controls (ICs) (two with major breakpoint region [MBR] and two with minor cluster region [mcr] rearrangements). The kinetics of amplification of ICs paralleled those of bcl-2/IgH rearranged genomic DNA. When used in a QC-PCR assay, these ICs were accurate at a 0.2-log level and provided reproducible results, as shown by low intrarun and interrun variability. An excellent correlation between predicted and observed lymphoma cell content (r = .99) was observed over a range of at least 5 logs of rearranged cells. This approach was used to measure involvement by NHL cells at the time of progenitor cell harvest in 37 autologous transplant patients. The number of bcl 2/IgH rearranged cells in BM, PB, and mobilized PB (mPB) was found to vary from 1 to 1.1 x 10(5) per million cells. The number of lymphoma cells present in BM was significantly higher than in PB (P = .0001), with a median difference in lymphoma cell content between BM and PB of 0.48 log of cells (range, -0.7 to 5 logs). In contrast, we found no difference in the concentration of bcl-2/IgH rearranged cells present in BM versus PB progenitor cells mobilized with cyclophosphamide and granulocyte colony-stimulating factor (G-CSF) (mPB) (P = .57). In conclusion, the QC-PCR assay described in this study could measure accurately and reproducibly the number of bcl-2/IgH rearranged cells among normal cells. Differences in levels of contamination by lymphoma cells between BM and PB were of less than one log (10-fold), and no differences in lymphoma cell concentrations were observed between BM and mobilized PB. As more cells are usually infused with mPB than with BM grafts, mPB progenitor cell grafts may actually be associated with higher levels of contamination by lymphoma cells. Furthermore, this QC-PCR assay should provide an important tool to assess the prognostic impact of lymphoma cell burden both in progenitor cell grafts and in vivo. PMID- 9414304 TI - Impaired induction of the CD28-responsive complex in granulocyte colony stimulating factor mobilized CD4 T cells. AB - Use of the CD28/B7 costimulatory signal for T-cell activation was analyzed in granulocyte colony-stimulating factor (G-CSF) mobilized peripheral blood mononuclear cells (G-PBMCs) and in peripheral blood mononuclear cells obtained before administration of G-CSF (preG-PBMCs). CTLA4Ig inhibition of OKT3 stimulated proliferation was significantly lower in G-PBMCs compared with preG PBMCs (39.9% +/- 5.6% and 72.2% +/- 5.4%, respectively; P < .001). Furthermore, as shown in electrophoretic mobility-shift assays, the inducible level of the T cell transcription factor CD28 responsive complex (CD28RC) was suppressed in CD4 cells derived from G-PBMC. However, depletion of CD14 cells from G-PBMCs restored CD28RC induction to normal levels. Taken together, these findings suggest that the large number of CD14 monocytes in G-PBMCs may limit T-cell responsiveness by suppressing the induction of the CD28RC. PMID- 9414305 TI - In vitro and in vivo evidence that ex vivo cytokine priming of donor marrow cells may ameliorate posttransplant thrombocytopenia. AB - Thrombocytopenia is typically observed in patients undergoing hematopoietic stem cell transplantation. We hypothesized that delayed platelet count recovery might be ameliorated by increasing the number of megakaryocyte colony- forming units (CFU-Meg) in the hematopoietic cell graft. To test this hypothesis, we evaluated cytokine combinations and culture medium potentially useful for expanding CFU-Meg in vitro. We then examined the ability of expanded cells to accelerate platelet recovery in an animal transplant model. Depending on the cytokine combination used, we found that culturing marrow CD34+ cells for 7 to 10 days in serum-free cultures was able to expand CFU-Meg approximately 40 to 80 times over input number. Shorter incubation periods were also found to be effective and when CD34+ cells were exposed to thrombopoietin (TPO), kit ligand (KL), interleukin-1alpha (IL-1alpha), and IL-3 in serum-free cultures for as few as 48 hours, the number of assayable CFU-Meg was still increased approximately threefold over input number. Of interest, cytokine primed marrow cells were also found to form colonies in vitro more quickly than unprimed cells. The potential clinical utility of this short-term expansion strategy was subsequently tested in an in vivo animal model. Lethally irradiated Balb-C mice were transplanted with previously frozen syngeneic marrow mononuclear cells (10(6)/mouse), one tenth of which (10(5)) had been primed with [TPO, KL, IL-1a, and IL-3] under serum-free conditions for 36 hours before cryopreservation. Mice receiving the primed frozen marrow cells recovered their platelet and neutrophil counts 3 to 5 days earlier than mice transplanted with unprimed cells. Mice which received marrow cells that had been primed after thawing but before transplantation had similar recovery kinetics. We conclude that pretransplant priming of hematopoietic cells leads to faster recovery of all hematopoietic lineages. Equally important, donor cell priming before transplant may represent a highly cost-effective alternative to constant administration of cytokines during the posttransplant recovery period. PMID- 9414306 TI - Chronic hepatitis C virus and gastric MALT lymphoma. PMID- 9414307 TI - Chronic hepatitis C virus and gastric MALT lymphoma PMID- 9414308 TI - Congenital erythropoietin-dependent erythrocytosis responsive to theophylline treatment. PMID- 9414309 TI - Clinical interaction between grapefruit juice and cyclosporine: is there any interest for the hematologists? PMID- 9414310 TI - An erythroid-specific exon is present in the human hexokinase gene. PMID- 9414311 TI - A new factor V gene polymorphism (His 1254 Arg) present in subjects of african origin mimics the R2 polymorphism (His 1299 Arg) PMID- 9414312 TI - Differentiating juvenile myelomonocytic leukemia from infectious disease. PMID- 9414315 TI - Sequence ready-or not? PMID- 9414313 TI - Hereditary hyperferritinemia-cataract syndrome: two novel mutations in the L ferritin iron-responsive element. PMID- 9414316 TI - Man to mouse--lessons learned from the distal end of the human X chromosome. PMID- 9414314 TI - Beta-spectrin Promiss-ao: a translation initiation codon mutation of the beta spectrin gene (ATG --> GTG) associated with hereditary spherocytosis and spectrin deficiency in a Brazilian family. PMID- 9414317 TI - WebWise: the Washington University Genome Sequencing Center's web site. PMID- 9414318 TI - Zooming in on the human-mouse comparative map: genome conservation re-examined on a high-resolution scale. AB - Over the past decade, conservation of genetic linkage groups has been shown in mammals and used to great advantage, fueling significant exchanges of gene mapping and functional information especially between the genomes of humans and mice. As human physical maps increase in resolution from chromosome bands to nucleotide sequence, comparative alignments of mouse and human regions have revealed striking similarities and surprising differences between the genomes of these two best-mapped mammalian species. Whereas, at present, very few mouse and human regions have been compared on the physical level, existing studies provide intriguing insights to genome evolution, including the observation of recent duplications and deletions of genes that may play significant roles in defining some of the biological differences between the two species. Although high resolution conserved marker-based maps are currently available only for human and mouse, a variety of new methods and resources are speeding the development of comparative maps of additional organisms. These advances mark the first step toward establishment of the human genome as a reference map for vertebrate species, providing evolutionary and functional annotation to human sequence and vast new resources for genetic analysis of a variety of commercially, medically, and ecologically important animal models. PMID- 9414319 TI - The comparative genomic structure and sequence of the surfeit gene homologs in the puffer fish Fugu rubripes and their association with CpG-rich islands. AB - The puffer fish Fugu rubripes (Fugu) has a compact genome approximately one seventh the size of man, mainly owing to small intron size and the presence of few dispersed repetitive DNA elements, which greatly facilitates the study of its genes at the genomic level. It has been shown previously that, whereas the Surfeit genes are tightly clustered at a single locus in mammals and birds, the genes are found at three separate loci in the Fugu genome. Here, Fugu gene homologs of all six Surfeit genes (Surf-1 to Surf-6) have been cloned and sequenced, and their gene structure has been compared with that of their mammalian and avian homologs. The predicted protein products of each gene are well conserved between vertebrate species, and in most cases their gene structures are identical to their mammalian and avian homologs except for the Fugu Surf-6 gene, which was found to lack an intron present in the mouse gene. In addition, we have identified conserved regulatory elements at the 5' and 3' ends of the Surf-3/rpL7a gene by comparison with the mammalian and chicken Surf 3/rpL7a gene homologs, including the presence of a polypyrimidine tract at the extreme 5' end of this ribosomal protein gene. The Fugu Surfeit gene homologs appear to be associated with CpG-rich islands, like the Surfeit genes in higher vertebrates, but these Fugu CpG islands are similar to the nonclassical islands characteristic of other fish species. Our observations support the use of the Fugu genome to study vertebrate gene structure, to predict the structure of mammalian genes, and to identify vertebrate regulatory elements. [The sequence data described in this paper have been submitted to the data library under accession nos. Y15170 (Surf-2, Surf-4), Y15171 (Surf-3, Surf-1, Surf-6), and Y15172 (Surf-5.)] PMID- 9414321 TI - Genome mapping by fluorescent fingerprinting. AB - The construction of sequence-ready maps of overlapping genomic clones is central to large-scale genome sequencing. We have implemented a method for fluorescent fingerprinting of bacterial clones to assemble contig maps. The method utilizes three spectrally distinct fluorescently tagged dideoxy ATPs to specifically label the HindIII termini in HindIII and Sau3AI restriction digests of clones that are multiplexed prior to electrophoresis and data collection. There is excellent reproducibility of raw data, improved resolution of large fragments, and concordance between the results obtained using this and the equivalent radioactive protocol. This method also allows detection of smaller overlaps between clones when compared to the analysis of restriction digests on nondenaturing agarose gels. PMID- 9414320 TI - A first-generation whole genome-radiation hybrid map spanning the mouse genome. AB - We have assembled a first-generation anchor map of the mouse genome using a panel of 94 whole-genome-radiation hybrids (WG-RHs) and 271 sequence-tagged sites (STSs). This is the first genome-wide RH anchor map of a model organism. All of the STSs have been previously localized on the genetic map and are located 8.8 Mb apart on average. This mouse WG-RH panel, known as T31, has an average retention frequency of 27.6% and an estimated potential resolution of 145 kb, making it a powerful resource for efficient large-scale expressed sequence tag mapping. [All of the mapping data for the maps presented here have been deposited at the Research Genetics, Inc., web site and can be freely accessed and downloaded at http://www.resgen.com/.] PMID- 9414322 TI - The complete set of predicted genes from Saccharomyces cerevisiae in a readily usable form. AB - Nearly all of the open reading frames (ORFs) of the yeast Saccharomyces cerevisiae have been synthesized by PCR using a set of approximately 6000 primer pairs. Each of the forward primers has a common 22-base sequence at its 5' end, and each of the back primers has a common 20-base sequence at its 5' end. These common termini allow reamplification of the entire set of original PCR products using a single pair of longer primers-in our case, 70 bases. The resulting 70 base elements that flank each ORF can be used for rapid and efficient cloning into a linearized yeast vector that contains these same elements at its termini. This cloning by genetic recombination obviates the need for ligations or bacterial manipulations and should permit convenient global approaches to gene function that require the assay of each putative yeast gene. PMID- 9414323 TI - Cloning-free PCR-based allele replacement methods. AB - Efficient homologous recombination permits the directed introduction of specific mutations into the yeast genome. Here we describe a cloning-free, PCR-based allele replacement method that simplifies allele transfer between yeast strains. The desired allele from one strain is amplified by PCR, along with a selectable/counterselectable marker. After transformation, the resident allele in the target strain is replaced by creating a duplication of the new allele. Selection for direct repeat recombinants results in a single copy of the new allele in the target strain. Specifically, the desired allele is amplified by PCR with a pair of adaptamers, which are chimeric oligonucleotides that are used to amplify the allele and differentially tag its 5' and 3' ends. These tags allow the directed fusion to two different, but overlapping, regions of an appropriately tagged selectable/counterselectable marker after a second round of PCR amplification. Following cotransformation of the two fusion fragments into yeast, homologous recombination efficiently generates a duplication of the amplified allele flanking the intact selectable marker in the genome. After counterselection, only the desired allele is retained as a result of direct repeat recombination. A simple modification of this method allows the creation of de novo mutations in the genome. PMID- 9414324 TI - The effect of age on the association between body-mass index and mortality. AB - BACKGROUND: The effect of age on optimal body weight is controversial, and few studies have had adequate numbers of subjects to analyze mortality as a function of body-mass index across age groups. METHODS: We studied mortality over 12 years among white men and women who participated in the American Cancer Society's Cancer Prevention Study I (from 1960 through 1972). The 62,116 men and 262,019 women included in this analysis had never smoked cigarettes, had no history of heart disease, stroke, or cancer (other than skin cancer) at base line in 1959 1960, and had no history of recent unintentional weight loss. The date and cause of death for subjects who died were determined from death certificates. The associations between body-mass index (defined as the weight in kilograms divided by the square of the height in meters) and mortality were examined for six age groups in analyses in which we adjusted for age, educational level, physical activity, and alcohol consumption. RESULTS: Greater body-mass index was associated with higher mortality from all causes and from cardiovascular disease in men and women up to 75 years of age. However, the relative risk associated with greater body-mass index declined with age. For example, for mortality from cardiovascular disease, the relative risk associated with an increment of 1 in the body-mass index was 1.10 (95 percent confidence interval, 1.04 to 1.16) for 30-to-44-year-old men and 1.03 (95 percent confidence interval, 1.02 to 1.05) for 65-to-74-year-old men. For women, the corresponding relative risk estimates were 1.08 (95 percent confidence interval, 1.05 to 1.11) and 1.02 (95 percent confidence interval, 1.02 to 1.03). CONCLUSIONS: Excess body weight increases the risk of death from any cause and from cardiovascular disease in adults between 30 and 74 years of age. The relative risk associated with greater body weight is higher among younger subjects. PMID- 9414325 TI - A comparison of the early outcome of acute myocardial infarction in women and men. The Third International Study of Infarct Survival Collaborative Group. AB - BACKGROUND: In previous studies, unadjusted comparisons of mortality and major morbidity after acute myocardial infarction have generally indicated that women have a poorer outcome than men. Much larger studies are needed, with more complete adjustment for coexisting conditions, to determine whether this difference is explained by the older age of the women studied or by the presence of other unfavorable prognostic factors, or both. METHODS: As part of the Third International Study of Infarct Survival (ISIS-3), information was collected on deaths during days 0 to 35 and on major clinical events during hospitalization up to day 35 for 9600 women and 26,480 men with suspected acute myocardial infarction who were considered to have a clear indication for fibrinolytic therapy. We compared the outcome among women and men, first without adjustment, then with adjustment for age, and finally with adjustment for other recorded baseline characteristics by means of multiple logistic regression. RESULTS: The unadjusted odds ratio for death among women as compared with men was 1.73 (95 percent confidence interval, 1.61 to 1.86). The women were significantly older than the men, and after adjustment for age the odds ratio was reduced markedly to 1.20 (95 percent confidence interval, 1.11 to 1.29). Adjustment for other differences in base-line clinical features further reduced the odds ratio to 1.14 (95 percent confidence interval, 1.05 to 1.23). Excesses in other major clinical events among women were generally reduced to a similar extent by adjustment. CONCLUSIONS: It seems likely that there is at most only a small independent association between female sex and early mortality and morbidity after suspected acute myocardial infarction. PMID- 9414326 TI - Monitoring women at risk for preterm labor. AB - BACKGROUND: Preterm birth is a major cause of perinatal morbidity and mortality. Whether the rate of preterm birth can be reduced by frequent contact between nurses and pregnant women or home monitoring of uterine activity is not known. METHODS: We randomly assigned 2422 pregnant women with known risk factors for preterm labor (including 844 women who were pregnant with twins) to receive education and to have one of the following: weekly contact with a nurse, daily contact with a nurse, or daily contact with a nurse and home monitoring of uterine activity. The nurses elicited the women's own assessments of their symptoms and signs of preterm labor. The primary end point was the incidence of birth at less than 35 weeks' gestation. Secondary end points included cervical status at the time preterm labor was diagnosed and birth weight. RESULTS: There were no significant differences among the groups in the incidence of birth at less than 35 weeks (14 percent in the weekly-contact group, 13 percent in the daily-contact group, and 14 percent in the home-monitoring group), in the mean amount of cervical dilatation at the time preterm labor was diagnosed (1.8 cm, 1.5 cm, and 1.4 cm, respectively), or in such neonatal outcomes as birth weights of less than 1500 g or less than 2500 g. However, daily contact with a nurse increased the mean number of unscheduled visits to obstetricians (1.2 in the weekly-contact group, 1.8 in the daily-contact group, and 2.3 in the home monitoring group) and the proportion of women who received prophylactic tocolytic drugs (12 percent, 14 percent, and 19 percent, respectively). CONCLUSIONS: Women who have daily contact with a nurse, with or without home monitoring of uterine activity, have no better pregnancy outcomes than women who have weekly contact with a nurse. PMID- 9414327 TI - A population-based study of seizures after traumatic brain injuries. AB - BACKGROUND: The risk of seizures is increased after traumatic brain injury, but the extent and duration of the increase in risk are unknown. The purpose of this study was to identify the characteristics of brain injuries that are associated with the development of seizures. METHODS: We identified 4541 children and adults with traumatic brain injury (characterized by loss of consciousness, post traumatic amnesia, or skull fracture) in Olmsted County, Minnesota, during the period from 1935 through 1984. Injuries were classified as mild (loss of consciousness or amnesia lasting less than 30 minutes), moderate (loss of consciousness for 30 minutes to 24 hours or a skull fracture), or severe (loss of consciousness or amnesia for more than 24 hours, subdural hematoma, or brain contusion). We compared the incidence of new unprovoked seizures in this cohort with population rates, using standardized incidence ratios and Cox proportional hazards analysis. RESULTS: The overall standardized incidence ratio was 3.1 (95 percent confidence interval, 2.5 to 3.8). The standardized incidence ratio was 1.5 (95 percent confidence interval, 1.0 to 2.2) after mild injuries but with no increase over the expected number after five years, 2.9 (95 percent confidence interval, 1.9 to 4.1) after moderate injuries, and 17.0 (95 percent confidence interval, 12.3 to 23.6) after severe injuries. In the multivariate analysis, significant risk factors for later seizures were brain contusion with subdural hematoma, skull fracture, loss of consciousness or amnesia for more than one day, and an age of 65 years or older. CONCLUSIONS: The increased risk of seizures after traumatic brain injury varies greatly according to the severity of the injury and the time since the injury. PMID- 9414328 TI - Images in clinical medicine. Poor skin turgor. PMID- 9414329 TI - Management of life-threatening acid-base disorders. First of two parts. PMID- 9414330 TI - Quinidine. PMID- 9414331 TI - Clinical problem-solving. A hidden agenda. PMID- 9414333 TI - Prevention of preterm birth. PMID- 9414332 TI - Losing weight--an ill-fated New Year's resolution. PMID- 9414334 TI - Reflections of a lake. PMID- 9414335 TI - The prevention of back injuries in Swedish health care--a comparison between two models for action-oriented feedback. PROSA Study Group. AB - This paper considers two different models for action-oriented feedback designed to prevent back injuries in Swedish health care. Feedback was provided by physiotherapists/ergonomists from Occupational Health Services. The models tested were feedback to work groups, and feedback solely to supervisors. Both generated a considerable number of accident-prevention proposals. Proposals of supervisors were implemented to a greater extent. Supervisors tended to advocate training, while work groups were more concerned with improved technical aids. Otherwise, only minor differences were detected between the models in application. The active involvement promoted by feedback, in particular to work groups, was considerable. PMID- 9414337 TI - The validation of three human reliability quantification techniques--THERP, HEART and JHEDI: Part II--Results of validation exercise. AB - This is the second of three papers dealing with the validation of three Human Reliability Assessment (HRA) techniques. The first paper introduced the need for validation, the techniques themselves and pertinent validation issues. This second paper details the results of the validation study carried out on the Human Reliability Quantification techniques THERP, HEART and JHEDI. The validation study used 30 real Human Error Probabilities (HEPs) and 30 active Human Reliability Assessment (HRA) assessors, 10 per technique. The results were that 23 of the assessors showed a significant correlation between their estimates and the real HEPs, supporting the predictive accuracy of the techniques. Overall precision showed 72% (60-87%) of all HEPs to be within a factor of 10 of the true HEPs, with 38% of all estimates being within a factor of three of the true values. Techniques also tended to be pessimistic rather than optimistic, when they were imprecise. These results lend support to the empirical validity of these three approaches. PMID- 9414336 TI - The impact of keyboard design on comfort and productivity in a text-entry task. AB - Concerns have arisen that the keyboard is a causal factor in the development of work-related musculoskeletal disorders (WRMDs) among video display terminal (VDT) operators. A number of alternative keyboard designs have been developed with altered geometry in an effort to improve comfort in keyboard operation. However, few data are available to substantiate whether these new keyboard designs are actually effective in reducing discomfort and musculoskeletal problems in users. The purpose of this study was to provide data on the efficacy of certain alternative keyboard design features (e.g. splitting the keyboard in half, and laterally inclining the keyboard halves) in reducing fatigue and musculoskeletal discomfort among keyboard operators. The study also explored the effects of these design features on performance. Fifty subjects performed a text-entry task for one day on a standard keyboard, then were assigned to one of five keyboard conditions for an evaluation period of two days (i.e. 10 subjects/condition). Outcome measures included performance (i.e. keystrokes/h, errors/h) and self report measures of discomfort and fatigue. The results indicated an initial decline in productivity when subjects began typing on two of the alternative keyboards, but these productivity losses were recovered within the two-day evaluation period. The results also indicated no significant differences between keyboard conditions in discomfort and fatigue. These results suggest a minimal impact of the keyboard design features examined in this study on productivity, comfort and fatigue, at least after two days of exposure. PMID- 9414338 TI - The validation of three human reliability quantification techniques--THERP, HEART and JHEDI: Part III--Practical aspects of the usage of the techniques. AB - This is the third paper in a series of three dealing with the detailed investigation of the empirical validity of three human reliability assessment (HRA) techniques. The first paper introduced the need for validation and specified the three techniques most requiring validation. The second paper detailed the results of an extensive independent validation experiment. This experimental validation involved 30 UK assessors using the techniques THERP, HEART and JHEDI (10 assessors per technique) to estimate the human error probabilities (HEPs) for 30 nuclear power and reprocessing (NP&R) tasks. The results for all three techniques were positive in terms of significant correlations, and general precision levels of 72% of all HEP estimates within a factor of 10 of the true value (unknown to the assessors). These results lend support to the empirical validity of these techniques in particular, and to HRA in general. However, the results were not all positive. In particular the consistency of usage of the techniques was variable. Additionally, subjects were generally not good at knowing their own uncertainty, i.e. they were not able to accurately predict when they were accurate nor when they were inaccurate. This desirable parameter is known as calibration, and the results from the validation suggested that subjects were not well-calibrated. This paper aims to determine how consistency of usage can be improved and to discern whether certain task types are, in practice, not well-assessed by the techniques, and hence are effectively currently beyond these techniques' abilities. Such information is aimed at aiding the HRA practitioner, or the ergonomist, interested in using these techniques. Recommendations for improving calibration are also discussed in this paper. A subsidiary but important focus of this paper is of a more fundamental nature, and of more general interest to the ergonomist. It concerns the validity of the techniques from an error reduction perspective. Currently these techniques may be used to identify how to reduce error probability, which is generally (in the qualitative sense) within the domain of ergonomics. One major mechanism for HRA-based error reduction is the utilisation of Performance Shaping Factor (PSF) information. This paper considers the validity of these PSF as ergonomics constructs. Drawing results from the validation exercise, it is seen how different PSF can be applied to the same scenario and can result in the same error probability, but will result in different error reduction guidance. It is therefore recommended that error reduction guidance must be based on a composite analysis of the results of the task, error identification and quantification analyses, with most weighting given to the qualitative analyses. PMID- 9414339 TI - Forearm muscular load and wrist angle among automobile assembly line workers in relation to symptoms. AB - Electromyographic activity (EMG) from m. flexor carpi radialis (FCR) and m. extensor carpi radialis longus (ECRL) of the right forearm was recorded together with wrist angles in the flexion/extension and radial/ulnar plane in 20 healthy automobile assembly line workers during work. Eleven of these were randomly recruited from assembly stations with a low prevalence of subjective wrist/forearm symptoms (LPS), while the rest came from stations with a high prevalence of symptoms (HPS). The main EMG finding was a clear difference in activation pattern between flexors and extensors. ECRL was activated more statically, while FCR had a more dynamic pattern with more pauses but also higher peak loads. The main wrist angle finding was a difference in angle distribution as well as absolute angular velocity in the radial/ulnar plane between LPS and HPS work stations. Workers in HPS stations worked longer times in a more ulnar deviated hand position and had higher absolute deviation angular velocity compared to LPS workers, indicating ulnar deviation as a risk factor. Generally, ulnar deviation from a neutral position was more frequent than angular displacement in the flexion/extension plane. PMID- 9414341 TI - Ergonomic redesign of the electric guitar. AB - The present study deals with the redesign of the electric guitar, considering ergonomic criteria. Difficulties met by novice musicians, neuro-muscular fatigue caused by guitar playing and occupational diseases occurring to professional guitarists, justify this study. Characteristics of existing electric guitar models which add to musician's fatigue or difficulty, were identified. A number of ergonomic requirements were then derived. The redesign process tried to satisfy these requirements, considering at the same time musical requirements and technical constraints. A comparative evaluation of the designed ergonomic guitar with three existing electric guitar models, showed that although the designed electric guitar preserves the main features of the instrument, it achieves better user fit. PMID- 9414340 TI - A neural network-based system for classification of industrial jobs with respect to risk of low back disorders due to workplace design. AB - Despite many years of research efforts, the occupational exposure limits of different risk factors for development of low back disorders (LBDs) have not yet been established. One of the main problems in setting such guidelines is the limited understanding of how different risk factors of LBDs interact in causing the injury, as the nature and mechanism of these disorders are relatively unknown phenomena. The task of an industrial ergonomist is complicated because the potential risk factors that may contribute to the onset of LBDs interact in a complex way, and require an analyst to apply elaborate data measurement and collection techniques for a realistic job analysis. This makes it difficult to discriminate well between the jobs that place workers at high or low risk of LBDs. The main objective of this study was to develop an artificial neural network-based diagnostic system which can classify industrial jobs according to the potential risk for low back disorders due to workplace design. Such a system could be useful in hazard analysis and injury prevention due to manual handling of loads in industrial environments. The results show that the developed diagnostic system can successfully classify jobs into the low and high risk categories of LBDs based on lifting task characteristics. PMID- 9414342 TI - Anthropometric data of Indian farm workers--a module analysis. AB - Anthropometric data of Indian farm workers have been compared with various other ethnic groups. The analysis indicates that there is a significant difference in the body dimensions which must be taken into account in designing farm equipment. PMID- 9414344 TI - Shoulder-arm muscle load and performance during control operation in forestry machines. Effects of changing to a new arm rest, lever and boom control system. AB - Three work station design studies in forestry machines were carried out with regard to shoulder/arm muscle load. (1) In an arm rest study, a movable (in the sagittal plane) arm rest was compared with a fixed one; (2) in a lever study, a mini lever with a small deflection was compared with an ordinary hand lever; (3) in a boom control study, a computer controlled boom system was compared with an ordinary boom system. In the arm rest study eight male subjects carried out lever operations in the laboratory. In the lever and boom studies, 15 male forestry students operated forwarders. Muscle load, cycle time, skill, perceived exertion, heart rate and preferred set up were recorded. The muscle loads on the upper trapezius, infraspinatus, extensor carpi ulnaris and flexor carpi radialis were recorded by electromyography. With mini lever and movable arm rest the upper trapezius load was reduced with preserved or decreased cycle time, although the grand mean of the 'static' load level was only about 2% of maximal voluntary contraction (MVC) and the effects of the new designs were small (about 0.7% MVC). The subjects preferred the lever/arm rest also giving the lowest muscle load. No differences between levers or arm rests were found in skill, perceived exertion or heart rate. It was not possible to draw any definitive conclusions regarding effects of the boom control system, due to technical problems. According to the literature, the duration of lever operation has increased considerably in forestry during recent years as part of a rationalization process. Partly due to this, it is suggested that the investigated work station improvements may not be sufficient to eliminate the risk for shoulder-neck disorders. PMID- 9414343 TI - Validation of Portable Ergonomic Observation (PEO) method using optoelectronic and video recordings. AB - The validity of the 'Portable Ergonomic Observation' method (PEO) was investigated against posture measurements based on continuous optoelectronic and video recordings made simultaneously with the observation. Work postures and actions during different frequently changing tasks were analyzed using both methods. In general, a high agreement between measured and observed data was achieved for the duration of clearly identifiable sustained postures and actions (such as repetitive movements of hands when typing), as well as for the frequency of clearly distinguishable actions (such as lifts). The agreement between observations and measurements was low for neck postures. In dynamic tasks the agreement was generally low. This was probably because of high levels of simultaneous information for the observer. No improvement in the agreement between measured and observed events could be achieved by leaving parts of the PEO categories unobserved at a time. Playback of video tapes to observe each category separately would increase the reliability of the observations but at the expense of increased time for the analysis. PMID- 9414345 TI - Effects of changes in work methods on musculoskeletal load. An intervention study in the trailer assembly. AB - The purpose of this case study was to identify the work cycles imposing high loads on the upper limb and low-back among trailer assembly workers for an ergonomic intervention. Several changes in work methods, tools and work organisation were implemented. The number of repetitions of fundamental work cycles and wrist posture were recorded from the video. Exposure imposed on the upper extremity in driving screws and drilling was computed based on experiments on force and time requirements. In lifting or carrying, the dose imposed on the low-back was computed based on the load moment on the L5/S1 disk and duration of the cycle. It was shown that after the intervention most workers worked less with deviated wrist and the cumulative exposure on the upper extremity was lower in furniture fixing tasks. Lifting with twisted torso was reduced. It was concluded that with relatively simple and low-cost solutions exposure to important risk factors of upper extremity and low-back disorders could be reduced. PMID- 9414346 TI - A user-centred design approach for introducing computer-based process information systems. AB - There has been an increasing tendency to use computer-based process information systems as the main interface through which operators interact with complex industrial systems. Although the new technology has produced greater hardware reliability and maintainability, the corresponding potential benefits for operability have not always been achieved. Automation has introduced new forms of design and operating errors. One of the major reasons for this problem has been the lack of human factors advice and user participation early in the design process. This paper discusses a user-centred design approach to increase operability and user acceptance of new technologies and working practices. Application of this approach in the context of a chemical plant indicates its promise, but also highlights the difficulties involved in gaining user participation and management commitment. PMID- 9414347 TI - Effects of rifle weight and handling length on shooting performance. AB - The effects of rifle design parameters on shooting performance were explored in this study. Twelve subjects, all right handed experienced marksmen, were recruited for the experiments. In the first part of the experiment, effects of rifle weight and handling length on aiming stability were examined in a simulated aiming exercise. Multiple responses of joint angles of the upper extremities, EMG data of selected muscles, center of pressure fluctuations, aiming point fluctuations and subjective preference were analyzed. It was shown that different rifle designs led to an alteration of rifle holding postures and muscle activation levels in order to maintain the system in balance, thereby affecting aiming stability. In the second part of the experiment, the relationship between aiming stability and shooting accuracy was examined in a live-fire study. Aiming fluctuations data of the same subject in the two experiments were highly correlated (r = 0.92-0.94, p < 0.0001). Significant high correlations between aiming fluctuations and shot group dispersions (r = 0.86-0.98, p < 0.0001) support the assertion that aiming stability and successful shooting performance are interrelated. A practical implication of rifle design trade-off was addressed in the paper. PMID- 9414348 TI - An electromyographic study of strength and upper extremity muscle activity in simulated meat cutting tasks. AB - Meat cutting has long been associated with a high incidence rate of upper extremity musculoskeletal disorders. This study examined upper extremity muscle activities and force exertion capabilities to identify postures which have potential for causing overexertion injuries. Fifteen subjects exerted force against a handle in postures similar to those observed in the meatpacking industry. Exertion level, direction of exertion, handle height, reach distance and grip type were varied. Activity in the posterior deltoid, biceps brachii, triceps brachii, extensor digitorum and flexor digitorum superficialis was monitored via surface electromyography (EMG). The ratio of normalized EMG activity to force produced during the exertion was computed for each muscle under each condition. The results showed that handle position had a significant effect on force exertion capability and the EMG/force ratio in all muscles. Force exertion capability was maximized, and the EMG/force ratio was generally minimized when participants pulled downward on a handle positioned at full arm's reach above the shoulder. For vertical cuts, force decreased and muscle activity generally increased as the handle height was lowered. For horizontal cuts, the full reach distance tended to allow greater force exertion with lower EMG/force ratios. The stab grip also tended to be associated with higher forces and lower EMG/force ratios than the slice grip. This study supports the premise that musculoskeletal stresses in meatpacking tasks can be altered through tool and workstation redesign. The data provided herein may be useful in selecting design modifications that reduce biomechanical stress on the upper extremities. PMID- 9414349 TI - Measurement variability in upper extremity posture among VDT users. AB - Hand and arm posture while keying is frequently mentioned as a risk factor for upper extremity musculoskeletal disorders (UEMSDs) among video display terminal (VDT) operators. However, many epidemiologic studies have not included measures of posture of VDT operators, in part, because of the difficulty of assessing posture rapidly and reliably among large numbers of subjects. For a single measure of posture to be useful for estimating dose-response relationships between posture and risk of UEMSDs, the within-subject variability of the postural measure must be smaller than the between-subject variability of the postural measure. In addition, the measure must be stable over time. We estimate the ratio of between- to within-subject variability for manual goniometry by measuring six postural angles on six occasions among 19 subjects using VDTs. For each postural angle, between-subject variability was substantially and statistically significantly larger than within-subject variability. Stability of postural measures over time was sufficient to justify a single postural measurement in epidemiologic studies. We conclude that manual goniometry can provide useful information about upper extremity posture among VDT users for use in epidemiologic studies of UEMSDs. PMID- 9414350 TI - Static anthropometric characteristics of students age range six-11 in Mazandaran province/Iran and school furniture design based on ergonomics principles. AB - This note describes the results of research into anthropometric measurement of children in Mazandaran province/Iran. Seventeen anthropometric measurements were taken from 1758 randomly selected subjects. Data obtained were used in school furniture design. PMID- 9414351 TI - Static anthropometric characteristics of Tehran University students age 20-30. AB - This study was conducted to establish an anthropometric database for Tehran University students (Iran). Twenty-eight dimensions from 179 students whose ages ranged from 20 to 30 were measured. Samples were randomly chosen. PMID- 9414352 TI - Variation in results of measurement repetition of human characteristics and activities. AB - The analysis of measurement variation in Ergonomics/Human Factors research reflects different approaches such as those prevalent in either the technical sciences or in the social sciences. The distinction is often a consequence of different types of measuring, for example measuring 'at' human beings as opposed to measuring 'through' human beings. In measuring 'at' human beings, as in anthropometrics, the measurement can be repeated several times. This allows the dispersion in results to be demonstrated via their standard deviation, as is usual in the technical sciences. In measuring 'through' human beings as, for example, in their performance in force exertion, a test-retest design is often adopted, i.e. a single repetition through several subjects in order to anticipate carry-over. In a number of research papers which were scrutinised, test-retest dispersion seems to have been analysed incompletely, ambiguously or incorrectly. In this paper a more appropriate way to specify within-subject dispersion is proposed, taking into account the dispersion patterning throughout the subjects. The relevance of studying this patterning is discussed, in terms of insight into human control, setting margins in design and for limiting the number of measurement repetitions per subject. PMID- 9414353 TI - Applying epidemiological principles to ergonomics: a checklist for incorporating sound design and interpretation of studies. AB - The primary purpose of this paper is to provide a checklist of scientific requirements necessary for the design of sound ergonomics studies. Ergonomics researchers will be able to use the checklist when designing a study and preparing it for publication. Practitioners can use the checklist to critically appraise study results, thereby having greater confidence when applying ergonomic recommendations to the workplace. A secondary purpose of the paper is to pilot the checklist on a sample of papers in the ergonomics literature and to assess its reliability. While there are checklists to assess the epidemiological rigour of studies, none have been adapted to address methodological issues in ergonomics. Two epidemiologists independently searched five ergonomics journals (Applied Ergonomics, Ergonomics, Human Factors, International Journal of Human Computer Interaction and Journal of Human Ergology) for research studies on VDT use and visual function published between 1990 and 1995. Twenty-one articles were reviewed. Each paper was scored according to the checklist. Overall, the reviewers found that the articles did not consistently fulfill some of the checklist criteria. An insufficient sample size was the most serious omission. Inter-rater reliability of the checklist was excellent for 11 of 14 items on the checklist (Kappa > 0.74), good for two items (Kappa between 0.40 and 0.74) and poor for one item. As ergonomics is gaining acceptance as an integral part of occupational health and safety, individuals in this field must be cognizant of the fact that study results are being applied directly to workplace procedures and design. It is incumbent upon ergonomists to base their work on a solid research foundation. The checklist can be used as a tool to improve study designs and so ultimately has implications for improving the fit between the worker and the work environment. PMID- 9414354 TI - An analysis of automatic teller machine usage by older adults: a structured interview approach. AB - It is often assumed that automatic teller machines (ATMs) are inherently easy to use and require no training. However, there is evidence to suggest that ATM users do experience difficulty when learning to use the system. The purpose of the present study was to conduct an in-depth analysis of ATM usage by older adults. Our approach consisted of telephone interviews followed by structured individual interviews. The goals were to understand the problems encountered by ATM users, to determine how ATMs might be better designed and to assess the training needs of older individuals. The phone interview data provide information about the relationships between age, sex and ATM usage within the adult sample, as well as information about why some people choose not to use ATMs. The structured interview data provide a more in-depth view of the concerns of both users and non users, and information about training needs. The training and design implications of the results are discussed. PMID- 9414355 TI - Effectiveness of elevator service signs: measurement of perceived understandability, willingness to comply and behaviour. AB - This research examines the effectiveness of four elevator service signs. The signs' purpose is to reduce delays for longer distance riders by dissuading people from using the elevator when they are only going up one floor or down two floors. Three of the four signs were described in Chapanis' (1965, Human Factors 7, 1-17) seminal treatise entitled 'Words, words, words ...': an original sign and two others that he suggested as possibly being better. The fourth was an enhanced sign incorporating human factor principles that were derived from research since Chapanis' article. The enhancements involved the use of colour, a signal word panel, icons/pictorial, and direct, explicit wording of the required behaviour. In Experiment 1, participants rated the understandability of each sign and their willingness to obey its instructions. The pattern of the means was the same for both questions. The original sign was rated lowest and the enhanced sign was rated highest, with the two other signs receiving intermediate ratings. In Experiment 2, the signs were placed on each floor of six multi-story buildings adjacent to the elevator call buttons. People's use of the elevators during the posting of each sign and during no-sign (control) periods was measured. The experimenter rode the elevators and counted the total number of passengers using the elevators as well as the number who rode up only one floor or down one or two floors (noncompliers). The new enhanced sign increased compliance compared to the other three signs and the no-sign period. These results suggest that design principles derived from recent research can help to promote comprehension, motivation and compliance behaviour to signs. PMID- 9414356 TI - An investigation of respiration while wearing back belts. AB - The research was conducted to evaluate the frequency of respiration during a repetitive lifting task when abdominal compression occurs from wearing a back belt. Three back belts were evaluated in this study: a nylon back belt, an inflatable back belt and an elastic vest. Analysis of the data revealed that the frequency of respiration increased while wearing the back belts at rest and while performing a repetitive lifting task. A statistically significant increase in the frequency of respiration was found while wearing the nylon back belt during the lifting task. PMID- 9414357 TI - Evaluating the motions of a semi-submersible platform with respect to human response. AB - The motions of a semi-submersible drilling platform have been evaluated so as to predict the effects on the comfort and activities of the crew. The horizontal motions at the drill floor exceeded the 'average threshold of perception' defined in International Standard 6897 (ISO 6897, 1984) by more than a factor of two; they were about half of the limit for the worst 10 min in five years for 'fixed offshore structures where work of a somewhat critical nature is carried out'. Other standards predict that the vertical motion would cause vomiting due to motion sickness in less than 5% of unadapted adults within the first 8h of exposure. The calculated probability of 'motion-induced interruptions' (loss-of balance events) caused by deck motion was negligible. Notwithstanding the above conclusions, it is considered that current standards are insufficient to predict the effects of the motions of ships and floating platforms on the activities of the crew. PMID- 9414358 TI - A field methodology for ergonomic analysis in occupational manual materials handling. AB - A methodology is presented for the 'on-site' evaluation of work-related physical activities. In a first session, video recordings were made of six industrial plant workers during their routine occupational tasks. Heart rate (HR) and subjective perception of effort were monitored. The video recordings were then analysed with appropriate software to determine the musculo-skeletal load ('Vision 3000', Promatek, Montreal) and to evaluate energy expenditure ('Energy', University of Michigan). The indirect estimates of energy expenditure were validated in a second session by monitoring the six subjects' oxygen consumption (VO2) during the same activities with a portable telemetric oxygen uptake analyser (Cosmed 'K2', Rome). No statistically significant differences were found between direct measurements of VO2 and the computerised estimates of energy expenditure. Biomechanical parameters obtained in the two sessions did not differ. Therefore, we conclude that the 'Energy' programme and the 'Vision 3000' program provide a fast and reliable profile of job requirements. PMID- 9414359 TI - Predicting pilot-error incidents of US airline pilots using logistic regression. AB - In a population of 70,164 airline pilots obtained from the Federal Aviation Administration, 475 males and 22 females had pilot-error incidents in the years 1986-1992. A simple chi-squared test revealed that female pilots employed by major airlines had a significantly greater likelihood of pilot-error incidents than their male colleagues. In order to control for age, experience (total flying hours), risk exposure (recent flying hours) and employer (major/non-major airline) simultaneously, the author built a model of male pilot-error incidents using logistic regression. The regression analysis indicated that youth, inexperience and non-major airline employer were independent contributors to the increased risk of pilot-error incidents. The results also provide further support to the literature that pilot performance does not differ significantly between male and female airline pilots. PMID- 9414360 TI - Physical load in ship maintenance: hazard evaluation by means of a workplace survey. AB - In a population of male workers in two ship maintenance companies (n = 32), a workplace survey was conducted in order to quantify their physical load. Postural load was measured by using the Ovako Working posture Analyzing System. During 7480 observations, working postures, exertion of force and working activities were recorded. Awkward postures of the back occurred in 38% of the worktime, stress on the neck/shoulder region due to one or both arms above shoulder level was present in 25% of the worktime. Forceful exertions during lifting, pushing and pulling activities sometimes exceeded published guidelines for manual material handling. Determinants of physical load could be identified and a hazard evaluation procedure was designed by applying rating schemes to weight various patterns of physical load. Ship maintenance work compared well with other strenuous occupations. Considering the high prevalence of back pain (80%) and neck/shoulder pain (60%), as well as the results of the observation method ergonomic improvements are warranted. Physical load can be reduced by several technical adaptations and applications, and by enlarging task rotation. PMID- 9414361 TI - A technique for recording and analysis of postural changes associated with thermal comfort. AB - The recording of posture has a long history in the study of ergonomics, but has generally been concerned with the assessment of mechanical strain on the human body. This paper extends the concern to the thermal implications of posture. A change in posture can change the effective body surface area available for heat exchange with the environment and therefore the metabolic rate per unit body surface area. This effect is systematised into a method of postural coding which reflects the extent of changes in effective body surface area available for heat exchange. The paper reports a brief assessment of the extent of the effect of posture in normal office work and the extent to which posture is temperature related. PMID- 9414362 TI - Maximum acceptable weight of lift by Chinese experienced male manual handlers. AB - This study uses a psychophysical methodology to determine the maximum acceptable weight of lift (MAWL) for 13 Chinese experienced male subjects, and examines the effects of variations in box size (including 300, 450 and 600 mm, sagittal distance from body) and lifting frequency (including one-time-maximum, 1, 4 and 6 lifts/min) on the MAWL, and the resulting responses [heart rate and rating of perceived exertion (RPE)]. The results are compared with prior studies and lead to the following conclusions: (1) Increases in both the box size and lifting frequency induced a significant decrease in MAWL, and a significant increase in RPE and heart rate. However, overall RPE ratings were not increased significantly with box size. (2) For Chinese inexperienced male subjects, the overall MAWL and heart rate were approximately 85% and 91% of their experienced male counterparts, respectively. The decrement in MAWL with Occidental subjects was about the same for both experienced and inexperienced groups; (3) The average MAWL values of Chinese subjects were smaller than those of Occidental subjects, and the average rate of decrease in MAWL with increasing of lifting frequency for the Chinese subjects was much sharper than in Occidental subjects; (4) There was no significant difference in overall heart rate and RPE ratings between the Chinese subjects and Occidental subjects. However, the body part RPE ratings were different from the previous studies. The present study found the most stressed body parts were the back and wrist, which may have been caused by the height of the table (760 mm) and the smaller body size of the Chinese subjects compared with the Occidental subjects. PMID- 9414363 TI - Computer mouse operation: is the left-handed user disadvantaged? AB - A survey of University of Melbourne student-use computers showed that 100% had the mouse installed on the right-hand side. An experiment was performed to determine if the left-handed user was disadvantaged by this arrangement. Times to move the cursor to targets of different sizes and distances showed that left handed users were not significantly disadvantaged and that, in accord with other tests, they were as good using their non-preferred hand as they were with their preferred hand. As expected, left-handers were superior to right-handed users when using their non-preferred hand. PMID- 9414364 TI - Evaluation of the impact of employee ergonomics training in industry. AB - This study examined the effects of three different types of ergonomics training methods upon employee knowledge, attitude and behavior. Employees within intact processing lines (N = 104) were randomized into four groups, one group serving as a control group. Pre- and post-test measures were implemented. Results showed training to have a significant effect upon knowledge of ergonomics. No significant differences were noted among all four groups according to empowerment and human factors measures. Training had a significant impact upon employee's job satisfaction, and the recognition and reporting of health hazards associated with their jobs. PMID- 9414365 TI - A questionnaire survey of the ergonomic problems associated with pipettes and their usage with specific reference to work-related upper limb disorders. AB - This study has considered the ergonomic problems associated with the use of pipettes through a questionnaire study of users. The study groups comprised an exposed (i.e. pipette users) and a non-exposed (i.e. non-users) cohort. Eighty questionnaire responses were returned by pipette users and 85 by non-users from six organisations; a response rate of approximately 55% for each of the study cohorts. The reported occurrence of elbow and hand complaints [using the general version of the Nordic musculoskeltal questionnaire (Kuorinka et al, 1987)] was significantly higher in the pipette user population as compared to the control population. There is an increase in the percentage of those reporting hand complaints as the duration of the working period involving continuous use of pipettes increases. Almost 90% of subjects in the longest exposure group (continuous use for more than 60 min) reported hand complaints. Users identified a number of features which made plunger operated pipettes more difficult to use: almost all of the female population who reported difficulties identified plunger operation as a design deficiency. Users also identified features of the general working environment which made the pipetting tasks more difficult. The study concludes that a number of work-related factors may affect the efficiency and comfort of staff performing laboratory tasks using pipettes. PMID- 9414366 TI - A comparison of the postures assumed when using laptop computers and desktop computers. AB - This study evaluated the postural implications of using a laptop computer. Laptop computer screens and keyboards are joined, and are therefore unable to be adjusted separately in terms of screen height and distance, and keyboard height and distance. The posture required for their use is likely to be constrained, as little adjustment can be made for the anthropometric differences of users. In addition to the postural constraints, the study looked at discomfort levels and performance when using laptops as compared with desktops. Statistical analysis showed significantly greater neck flexion and head tilt with laptop use. The other body angles measured (trunk, shoulder, elbow, wrist, and scapula and neck protraction/retraction) showed no statistical differences. The average discomfort experienced after using the laptop for 20 min, although appearing greater than the discomfort experienced after using the desktop, was not significantly greater. When using the laptop, subjects tended to perform better than when using the desktop, though not significantly so. Possible reasons for the results are discussed and implications of the findings outlined. PMID- 9414367 TI - Designing virtual selectors for surgeons. AB - Virtual Reality (VR) applications involve the use of manipulators or 'virtual tools' such as pointers, grippers and containers. Manipulation of such tools is a key feature of VR applications. Less attention has been directed towards designing the ways in which users select particular options or information, as they interact in the virtual world, by means of selectors. This is particularly important in the case of professional users, such as surgeons, who typically make heavy use of both selection and manipulation, switching frequently between the two in the course of their work. In this paper we describe the design and evaluation of two types of selector tool for use in medical VR applications. In this account, we provide details of the background behind the research and focus on design aspects of selector tools for use in a particular task context, that of medical surgery planning. PMID- 9414368 TI - Fastener systems on apparel for hemiplegic stroke victims. AB - Many stroke victims suffer hemiplegia. The type and location of apparel fasteners (e.g. buttons) are important to facilitate dressing for persons with hemiplegia. This study evaluated a series of garments with different fastener types and locations using hemiplegic subjects. The experimental design was a 3 x 3 + 1 factorial complete randomized block design with a control. Fastener type and location were independent variables. Dependent variables were: (1) time required to open and close the fasteners, (2) number of fasteners opened and closed, (3) and subjective responses to a questionnaire. The subjects for this study were ten stroke victims with hemiplegia. Results indicated significant differences for fastener type and location. These results should aid apparel manufacturers to provide more functional apparel for persons with hemiplegia due to stroke, as well as those responsible for selecting and/or helping to dress persons with hemiplegia due to stroke. PMID- 9414369 TI - Analysis of measurements of slip resistance of soiled surfaces on site. AB - The present study hinges on the use of the PFT (Portable Friction Tester) to assess the slip resistance of floors on site and aims to analyse measurements of slip resistance (602 measurements) that were carried out in 27 different firms on moistened, greasy or dirty floors. The contribution of different factors to the slipperiness of a given soiled surface can thus be identified or confirmed: quantity and viscosity of the soil, permeability and roughness of the floor, cleaning efficiency and uniformity. In particular, we observed that cleaning a soiled surface does not immediately lead to an increase in its slip resistance. The present study is exploratory in nature since so far no publication has been devoted to the analysis of measurements of slip resistance of floors in more than one firm. PMID- 9414370 TI - Technical note. Comparison between estimated worn clothing insulation and required calculated clothing insulation in moderately cold environments (0 degree C < or = ta < or = +15 degrees C). AB - Six female and 33 male workers of the food industry (16-55 years), divided into three groups according to climatic conditions at the workplaces, were monitored during a typical shift. Fourteen subjects worked continuously in 0-7 degrees C, 18 in 13-15 degrees C and seven moved frequently between these climatic areas. Mean metabolic rates, heart rates, rectal temperatures and skin temperatures at the trunk and at the feet were similar between each of the three groups. Considerable differences between estimated clothing insulations worn and calculated required clothing insulations (IREQneutral) were statistically analyzed. The results suggest that--apart from a limited overestimation--this discrepancy is mainly related to the difference between time-adjusted averages of metabolic rates of the single activities and the respective daily minimum suggesting the need for an adequate weighting for the metabolic rates, particularly if workers are at least temporarily exposed to air temperatures of more than 7-13 degrees C. PMID- 9414371 TI - Towards viable, useful and usable human factors design guidance. AB - This paper investigates the factors relevant to producing effective human factors design guidance, using the Engineering Data Compendium (EDC) as a research vehicle. A series of three exploratory experiments focusing on the factors that affect the usability, usefulness and viability of human factors handbooks was conducted. The results of these studies were interpreted in the context of the process by which the EDC was developed, leading to the following recommendations: (a) human factors guidance should be organized in a manner that is stepped in context; (b) human factors guidance should be based on an explicit requirements analysis; (c) the calibration of designers' perceptions of the cost of obtaining human factors information must be improved; (d) organizational policies must be changed to induce more effective information search behaviour. PMID- 9414372 TI - Testing the relative conspicuity of safety garments for New Zealand forestry workers. AB - The relative conspicuities of six test garments (fluorescent orange, green/red, high-contrast, fluorescent lime-yellow, white, black) were assessed to guide the selection process of a standard upper body safety garment to be used within the New Zealand logging industry. Six male and four female participants, aged between 18 and 26 years, volunteered to perform 10 trials each on a demanding, central (tracking) task, while peripherally searching colour slides for test garments displayed on each of eight positions, against a pine forest background typically found in the New Zealand forestry. During each trial, transparency luminance was first gradually increased from darkness to daylight and then decreased to darkness again within 180 s. A head mounted ASL eye tracking system (4000SU) recorded the eye line of gaze for each participant, enabling a rank order of detection to be obtained for the tested garments in each trial. The fluorescent lime-yellow, fluorescent orange and white test garments were detected earlier than any other test garments across all trials, and the fluorescent lime-yellow test garment was detected first with a higher frequency than the white test garment. It was concluded that while white may be the most visible colour in near darkness conditions, as it provides highest contrast, fluorescent lime-yellow stands out better in twilight and daylight conditions against the pine forest background. The results of this study led to the promotion of fluorescent lime yellow as the standard safety colour used in upper body garments within the New Zealand forest industry. PMID- 9414373 TI - Field study of subjective assessment of negative pressure half-masks. Influence of the work conditions on comfort and efficiency. AB - The aim of this study was to assess the effects of work conditions on the acceptability and efficiency of respiratory protective devices (RPD). The subjective evaluation of comfort, protection, respiratory and visual constraint, and the acceptable duration of wear of six RPDs against dust was achieved by 30 workers during their actual work. Metabolic rate was evaluated for each worker, and dry and wet air temperatures measured in the work area. RPDs objective protection factor was measured during each of the 180 test periods. In the conditions of this study, the acceptable duration of wear was about 1 h. This duration and the comfort parameters were reduced when the air temperature increased. The younger workers and/or smokers were less sensitive to mask discomfort. Objective protection factors of the RPDs are reduced under warmer conditions, and when the metabolic rate is low. Finally, the results of this study also show the poor capacity of standardized leakage tests to assess the objective respiratory protection of workers in the field. Some hypotheses which can explain this fact are discussed. PMID- 9414374 TI - The influence of physical fitness training on the manual material handling capability of women. AB - This study examined the influence of a generalized physical fitness training program on manual material handling (MMH) capability. Thirteen healthy women trained for 14 weeks, performing progressive resistance training three days per week and running with interval training two days per week. Subjects attended 85 +/- 6% of the sessions. Compared to values obtained before training, subjects increased the maximum mass they could lift from floor to knuckle height by 19% (68-81 kg, p < 0.001) and from floor to chest height by 16% (49-57 kg, p < 0.001). They improved by 17% their ability to lift 15 kg as many times as possible in 10 min(167-195 lifts, p < 0.001), while perception of effort (measured with the Borg Rating of Perceived Exertion) did not change. Total body mass did not change, but body fat mass was reduced by 9% (18.8-17.2 kg, p = 0.036) and fat-free mass increased by 6% (48.2-51.0 kg, p < 0.001). A short-term physical fitness program, conducted about 1 h per day, five days per week, can substantially improve women's MMH capability and provide favorable changes in body composition (increased fat-free mass and decreased body fat. PMID- 9414375 TI - Computer mouse or Trackpoint--effects on muscular load and operator experience. AB - The aim of the study was to evaluate four different modes of human-computer interaction. The modes were: use of the keyboard alone as input device, use of keyboard and mouse, use of keyboard and mouse with a three-dimensional arm support, and use of a keyboard with a Trackpoint device in its centre. Ten women and 10 men volunteered to participate. Questions asked were whether working in the different modes influenced shoulder and forearm muscular load differently during word processing, and how much strain on the neck, shoulder and arms subjects perceived in the different modes. Muscular load was studied with electromyography in three shoulder muscles and three forearm muscles. The subjects also rated the different modes in one questionnaire concerning perceived strain and in one concerning preference for any of the modes tested. Intra individual analysis for each muscle and mode showed two possible ways of decreasing the strain from computer mouse work on the shoulder muscles--either to use Trackpoint or to use the mouse combined with the movable arm support. However, both of these computer-interaction modes increased the muscular load in the hand and forearm. PMID- 9414376 TI - Differences in low back load between kneeling and seated working at ground level. AB - The effects of the load on the low back when working in a kneeling posture were compared to those when working on a chair designed to alleviate work load in harvesting radish. In 10 male subjects data regarding back muscle fatigue and length changes of the spine, and estimates of experienced discomfort were collected. The results show that for all three effects of back load working on the chair leads to lower levels as compared to working kneeling. However, back load and discomfort were lower when working on the chair, back discomfort still increased substantially during the work on the chair. It is thus a sub-optimal solution from an ergonomic point of view, but at present it could be recommended to allow for variation with the conventional working method. PMID- 9414377 TI - An ergonomic evaluation of a patient handling device: the elevate and transfer vehicle. AB - This study evaluated a Patient Handling Device (PHD) called the Elevate and Transfer Vehicle (ETV). The ETV works on the principle of leverage to transfer a patient from one seated position to another. Three types of product evaluation were used: expert appraisal; user trials; and performance tests. Expert appraisal was conducted by a panel of 11 people including an ergonomist, an industrial designer, two engineers, including one employed as an academic in a School of Mechanical, Manufacturing and Medical Engineering, and seven health professionals. The experts evaluated the ETV using a checklist and group discussions. They generally agreed that the advantages of the ETV tested were it's simplicity, the convenient position to adjust clothing for toileting and the need for only one carer. They noted comfort, security of straps, centre of gravity and manoeuvrability as the main areas for improvement. User trials consisted of nine male and nine female volunteer users assigned to carer/patient pairs. Following a training period, each user subjectively evaluated the ETV by structured interview. User trial results indicated ease of use, prevention of back injuries in carers and minimal body contact were advantages of the ETV. The main problems with using the ETV appeared to be the inadequate 'prop' and straps, the 'jolt' and lack of dignity for the patient. Several critical performance tests were conducted to determine compliance to Australian Standards for design. Areas of non-compliance included strength of frame and static stability. The findings suggest that most of the identified problems of the ETV could be overcome with minor design improvements. The general consensus of participants was to keep the design simple, maintain fast transfers and maintain the position of the patient to enable ease of clothing adjustment for toileting. PMID- 9414378 TI - Evaluating the effects of interface factors on the torque exertion capabilities of operating handwheels. AB - This study intends to assess factors affecting human torque exertion capabilities of operating valve handwheels (maximum volitional torque exertion of wrist radial/ulnar deviation, R/U MVTE). Forty student subjects (20 males and 20 females) participated in this study. In addition to gender and subject factors, gloves (one layer of cotton, two layers of cotton and rubber gloves), operating height (elbow, shoulder and overhead), handwheel size and shape were selected. Barehanded condition was also involved. The results indicate that all the main effects and the first order interactions were significant. The gloved R/U MVTEs were found to be greater than the barehanded R/U MVTE. For operating height, shoulder height gave the greatest R/U MVTE, followed by elbow and overhead heights. The handwheel diameters ranging from 75 to 95 mm for males and 65 to 80 mm for females were found to have the greater R/U shear force. The average R/U MVTE of operating valve handwheel for females was about 63% (3.8/6.05) of that of males. PMID- 9414379 TI - A method for dynamic measurement of the resistance to dry heat exchange by footwear. AB - Five different types of cold protective footwear have been tested with regard to their resistance to dry heat loss (i.e. the insulation) with a new electrically heated foot model. The model is able to simulate 'walking' movements in order to provide a more realistic simulation of wear conditions. Thermal insulation of shoes with and without a steel toe cap was the same. The insulating properties during simulated walking movements were 10-25% lower compared with static conditions. For two of the shoe models a significantly lower insulation value for the sole area was obtained when adding a weight of 30 kg. A significant difference could also be found between the insulation values of two different sizes of one of the models. Measurements with the standard method (EN 344) correlated well with the local insulation value of the sole part of the thermal foot. Correlation with the insulation value for the whole shoe was much less, variation was bigger and ranking in terms of cold protection differed between methods. The electrically heated foot model appears to provide a reproducible, accurate and more realistic method for measuring the insulation properties of shoes than EN 344. PMID- 9414380 TI - Perception of difficulties for the back related to assembly work: general findings and impact of back health. AB - The study objective was to describe the perceptions of airplane assemblers on job demand for the back and how back pain modulated these perceptions. One hundred and seventy-six workers answered two questionnaires concerning back pain and the perception of work related difficulties (work activities, work contexts, tools, work positions, efforts). Results show that positions and work contexts are perceived as greater sources of difficulty than efforts or dynamic activities. The duration of a given position is more important than its frequency. Back pain has a significant but complex impact on the perception of difficulty. Assemblers appear to integrate several factors when evaluating their difficulties as opposed to individual aspects, as it is often measured in ergonomic studies. The results have important implications for the measurement of ergonomic factors in the genesis of back pain and illustrates the potential for misclassification and biases in current epidemiologic studies. PMID- 9414381 TI - An analysis of human comfort when entering and exiting the rear seat of an automobile. AB - This paper describes a study of human motion and human comfort when entering and exiting the rear seat of an automobile. A simulator was used to test several possible door frame configurations, and various positions of the front and rear seats. Thirty-six human subjects were asked to enter and exit the simulator five times for each configuration and to answer a subjective questionnaire. The motion performed by each test subject was recorded by means of a VHS recorder and an ELITE motion measurement system. A statistical analysis was performed on the data from the questionnaires and comfort rankings were produced for the various configurations. The most influential design parameters were identified and iso comfort surfaces were defined and fitted which provided a simple means of quantifying the effect of one of the main parameters. PMID- 9414382 TI - Technical note. Development of a set of Korean manikins. AB - There is a difficulty in defining the 95% accommodation of a population for complex design problems when several body dimensions are involved. This study reports a method for constructing a family of manikins which are validated for population accommodation. They are analysed in terms of the interactions or relationships between the body dimensions' measurements. The proposed manikin family represents multivariate body measurements as a limited set of design alternatives. From the proposed method, a family of nine manikins is established to represent Korean males aged between 30 and 50 years to be used when determining workplace dimensions. PMID- 9414383 TI - Expression of cyclin D1, CDK4 and p27KIP1 is associated with the p16MTS1 gene status in human esophageal carcinoma cell lines. AB - p16MTS1/INK4A negatively regulates cell cycle progression by inhibiting the cyclin D/CDK4 complex that phosphorylates pRb. Frequent homozygous deletions of the p16 gene were recently found in various tumor cell lines. We examined the relationship between the genetic status of p16 and the expression of the cell cycle regulating molecules in human esophageal carcinoma cell lines. Out of eight human esophageal carcinoma cell lines, seven (67.5%) and six (75%) cell lines showed homozygous deletions of the p16 and p15 genes, respectively. All the p16 negative cell lines expressed high levels of cyclin D1, CDK4 and p27KIP1 proteins. Interestingly, the expression level of cyclin D1 was closely correlated to the levels of not only CDK4 but also p27KIP1 protein in p16-negative cell lines. Furthermore, all the p16-negative cell lines expressed Rb protein of approx 110 kDa which corresponds to the phosphorylated form, whereas the cell line with intact p15 and p16 genes did not express pRb. These results suggest that the expression of cyclin D1, CDK4, Rb and p27 is associated with the p16 gene status in esophageal carcinoma cell lines. Alternatively, loss of the p16 gene and subsequent over-expression of cyclin D1 and CDK4 might be involved in autonomous growth of esophageal carcinoma cells. PMID- 9414384 TI - Multidrug resistance-modifying components in human plasma with potential clinical significance. AB - P-Glycoprotein (P-gp) and multidrug resistance protein (MRP) are plasma membrane associated proteins which can confer multidrug resistance (MDR) to cancer cells by lowering the intracellular amount of drug. Although clinical trials with MDR reverting agents have been initiated, not much attention has been paid to blood components which may modulate the activity of P-gp or MRP. The present investigation was performed to identify and characterize blood components which may influence the drug content and the drug cytotoxicity of MDR cells. Human plasma, from healthy volunteers, was tested for its effects on the daunorubicin (DNR) accumulation and cytotoxicity in the MDR cell lines SW-1573/2R160 (2R160) and GLC4/ADR containing P-gp and MRP, respectively. The data were compared to the effects observed in wild-type cells. MDR-modifying plasma components were isolated by extraction procedures and characterized using ultrafiltration, high performance liquid chromatography (HPLC) and mass spectrometry. An increase in the proportion of plasma in the culture medium led to a reduction of the ratio between the DNR content of wild-type and corresponding MDR cells. At 100% plasma we observed an increase in the cellular DNR content of 2R160 cells, which was 10 30% (median 18%) of the maximum possible increase induced by well-known MDR reverting agents, such as verapamil (for GLC4/ADR cells: 10-20%, median 15%). The DNR cytotoxicity in MDR cells also increased with an increasing amount of plasma included in the culture media. There was neither an increase in the cellular DNR content nor an effect on the DNR cytotoxicity in wild-type cells. Plasma extract analysis by HPLC showed a major peak which increased the DNR content of MDR cells. The HPLC column retention time of this fraction was identical to that of a standard of cortisol and it was further confirmed to be cortisol using mass spectrometry. Moreover, inclusion of a standard of cortisol in culture media induced a similar effect. We analyzed the data for one of the plasma pools and found that blood cortisol was responsible for the MDR-modulating effect only for 35% of the effect of 100% plasma. Other plasma components were responsible for the remaining modulation effect on MDR cells. In conclusion, the DNR pumping activity of P-gp and MRP is inhibited by human plasma, resulting in 10-30% of the maximum possible increase in cellular drug content. Based on cellular pharmacokinetic calculations this percentage will most likely increase at clinical levels of drug resistance (reaching 40-50%). In one sample blood cortisol accounted for 35% of the effect of plasma on the DNR content in MDR 2R160 cells. These data show the need for additional studies to test plasma samples for their MDR modulating effects before the administration of MDR reverting agents in chemotherapy. The data suggest that the effectiveness of chemotherapeutic drugs may be enhanced when administered in accordance with the circadian peak of endogenous corticoids. PMID- 9414385 TI - In vivo reversibility of multidrug resistance by the MDR-modulator dexniguldipine (niguldipine derivative B859-35) and by verapamil. AB - The newly synthesized dihydropyridine derivative B859-35 was previously shown in vitro to be highly effective in reversing multidrug resistance (MDR) of P glycoprotein positive tumor cell lines, such as the adriamycin (ADR) resistant erythroleukemia F4-6RADR cells. In the current study B859-35 was investigated for its efficiency in reversing MDR in an in vivo tumor model for preclinical testing of MDR-modulators. F4-6RADR cells were injected into the right flank of nude mice while the parent cells were injected into the left flank. The animals were treated i.p. with ADR (9.0 mg/kg body weight) combined with B859-35 (5, 10, or 25 mg/kg) or, for comparison and validation, with verapamil (VRP) (75 mg/kg). The effects of ADR and the MDR-modulator combination were evaluated by histological morphometry of the tumors. While ADR alone was shown to be ineffective in resistant cells, the combinations of ADR + B859-35 as well as of ADR + VRP were highly active in reducing the number of viable cells in the resistant tumor nodule by 67 +/- 9% or by 53 +/- 11% of controls. This model provides evidence that even in vivo, MDR modulators can be effective in reversing drug resistance. In addition, it presents a potentially useful and rapid preclinical system for in vivo studies on the modification of drug resistance. PMID- 9414386 TI - Rates of development of methotrexate resistance in heterogeneous mouse mammary tumor cell cultures. AB - Our previous studies have indicated that the expression by tumor cells of sensitivity to chemotherapeutic drugs such as methotrexate can be affected by the presence of other tumor cells; thus, otherwise methotrexate-resistant cells may respond to that drug in the presence of methotrexate-sensitive cells. In order to determine whether tumor heterogeneity also affects the emergence of drug resistance, we measured the rate of development of methotrexate resistance in mixed monolayer cultures of three mammary tumor subpopulation lines (66, 168TFAR, 4T07) that differ in degree of sensitivity to methotrexate. Cultures were treated weekly with 80 nM or 200 nM methotrexate. Each individual cell line was re isolated from the mixture by passage in selective medium and then assayed for methotrexate sensitivity. Cultures of each of the three lines were treated and assayed in parallel. Few differences in the rate of development of methotrexate resistance were seen among cells from mixtures and cells cultured alone; line 4T07 appeared to become resistant somewhat more rapidly in mixtures. In untreated mixed cultures, line 66, the line least sensitive to methotrexate, gradually became dominant; this process was accelerated in treated cultures. One methotrexate-resistant subline from each parent cell line was tested to determine the mechanisms by which methotrexate resistance was increased. Two lines appeared to have increased levels of dihydrofolate reductase, and one exhibited decreased methotrexate transport as well. The third cell line had neither mechanism. Others have shown that tumor heterogeneity can act as a brake on the rate of development of new metastatic or immunogenic variants. Our data indicate that, at least in the model system we have tested, the rate of development of extrinsic drug resistance is substantially independent of pre-existing clonal diversity. PMID- 9414387 TI - Messenger RNA expression of resistance factors in human tumor cell lines after single exposure to radiation. AB - Resistance of tumor cells to chemotherapeutic drugs can not only be caused by treatment with antineoplastic agents but also by radiotherapy. The aim of this study was to analyze whether ionizing radiation can influence the mRNA expression of proteins which have been found to be involved in drug resistance of tumor cells. Human tumor cell lines (MCF-7, LXF and Sk-Mel) were treated with single doses of irradiation (5, 10 and 20 Gy). The expression of the resistance related proteins glutathione S-transferase-pi (GST-pi), topoisomerase II alpha (Topo II), thymidylate synthase (TS), O6-methylguanine-DNA-methyltransferase (MGMT), P glycoprotein (Pgp), glutathione peroxidase (GPX) multidrug resistance-associated protein (MRP) and also of the heat-shock protein 70 (HSP 70) were determined at the mRNA level during the time interval from 1.5 to 72 h post-irradiation and compared with their corresponding controls. We also examined whether a relationship exists between these proteins and the proliferative activity (histone 3, Ki-67, statin) of the cells. We found that exposure of MCF-7, LXF and Sk-Mel cells to ionizing radiation increases the expression of the mRNA of GST pi. Topo II, TS, HSP 70 and proliferation markers were also altered by exposure to ionizing radiation, but there was no common response of the three cell lines. No significant changes were observed in the expression of MGMT, Pgp, GPX and MRP after radiation treatment. Drug resistance tests revealed that irradiated MCF 7 cells were less sensitive to doxorubicin than non-irradiated control cells. Our results indicate that ionizing irradiation modifies the expression of some proteins involved in drug resistance and the response of MCF 7 cells to doxorubicin and may, therefore, play a role in clinical drug response. PMID- 9414388 TI - Ultrastructural changes related to multidrug resistance in CEM cells: role of cytoplasmic vesicles in drug exclusion. AB - The multidrug resistance phenotype is found to be frequently associated with the overexpression of proteins which lead to a decrease of drug accumulation within human tumor cells. A 170 kDa membrane glycoprotein which is related to the overexpression of the mdr1 gene is inserted in the plasma membrane and pumps the cytotoxic drugs out of the cells. The aim of this work was to study the morphological modifications of resistant CEM/VLB 100 cells relative to their parental drug-sensitive ones and the detection of the ultrastructural localization of P-glycoprotein at the cytoplasmic level. Using a scanning electron microscope, CEM resistant cells showed wide smooth protrusions while CEM sensitive cells showed microvilli and fine folds. With transmission electron microscopy, an enhanced secretory system was observed in CEM resistant cells: both electron transparent and electron opaque vesicles were associated with the Golgi system, revealed by wheat germ agglutinin-colloidal gold labelling. These vesicles were the binding site of C 219 and MRK 16 antimembrane glycoprotein antibodies, and some of them were determined to belong to the lysosomal system after PTA staining. These vesicles may be an additional way to decrease the cellular uptake of drugs in multidrug resistant cells. Moreover, some nuclear and nucleolar modifications were also observed. These observations show that MDR has wide morphological features which concern several organelles. PMID- 9414390 TI - The basis for somatic gene therapy of cancer. AB - A decade of advances in understanding of the molecular basis of sporadic and familial cancers has combined with developments in mammalian gene transfer technology to stimulate intensive research into the potential applications of somatic gene therapy for cancer. Somatic gene immunotherapy is already in progress to stimulate and direct the natural targeting capabilities of the immune system against the threat of disseminated residual disease. The association of a plethora of mutated tumor suppressor genes (p53, p16 BRCA1, BRCA2) with diverse cancers has also highlighted the potential of somatic gene therapy with wild-type versions of suppressor genes as an anti-cancer therapeutic modality either in its own right or in synergistic association with traditional anti-cancer therapies. The methodologies for gene transfer technology range from direct intravenous injection of naked modified DNAs to intravenous injection of liposome encapsulated DNAs or microsphere-bound DNAs. Recombinant retroviral and adenoviral vectors have natural transfection capabilities and display tropism for particular tissues that are of selective advantage against particular cancers. Liposomes display very high efficiencies of gene transfer with the advantages of successful transfer to a wide range of tissue types but their widespread systemic distribution offers problems in relation to selective targeting of tumor cells. The challenges to current gene transfer processes are much the same as that of other anti-cancer therapies: achieving selective targeting of cancer cells whilst optimizing dosages and minimizing the risk of collateral damage to healthy tissues. PMID- 9414389 TI - Relative bioavailability of 4,4'-dihydroxybenzophenone-2,4-dinitrophenylhydrazone (A-007) in rats and monkeys. AB - 4,4'-Dihydroxybenzophenone-2,4-dinitrophenylhydrazone (A-007) is being evaluated for its anticancer activities in melanoma, breast cancer, Kaposi's sarcoma and lymphoproliferative disorders. A single oral dose of 1 g/kg of A-007 in rats resulted in prolonged and low plasma levels, typically less than 150 ng/ml for several days. Similarly, a single oral dose of 5 g/kg of A-007 in monkeys resulted in prolonged and low plasma levels, typically less than 22 ng/ml for several days. Oral bioavailability data suggests that this is not an efficient mode of drug administration and availability diminishes as one progresses from rodents to primates (relative oral bioavailability 2%); thus suggesting an alternative form of drug delivery is required in higher species. A-007 is not detected in plasma after a 0.25% gel is applied topically to the skin daily for 28 days. Early clinical support the topical use of A-007 to treat cutaneous metastasis for human breast cancer. The present data further support a dermal approach for the use of A-007 to treat metastatic cutaneous cancers. PMID- 9414391 TI - Phase I trial of a 24-h continuous infusion of ifosfamide/mesna in acute lymphoblastic leukemia. AB - Based on previous reports suggesting that an intravenous (i.v.) continuous infusion of alkylating agents produced a significant response rate in acute lymphoblastic leukemia (ALL), a phase I trial of ifosfamide/mesna (IFO/MES) was conducted in 11 adult patients with relapsed ALL. IFO/MES were administered as a 24-h i.v. continuous infusion in doses ranging from 5 g/m2 to 9 g/m2; the courses of treatment were repeated every 3 weeks. Patients were examined for toxicity after every cycle and responses were carefully defined and evaluated. All 11 admitted patients were evaluable for toxicity and response. Myelosuppression was the dose limiting effect and it was dose-related. Microscopic and/of macroscopic hematuria was detected in four (11%) out of 36 cycles administered. Ifosfamide produced a positive biological response with a preliminary response rate of 45.4%. Ifosfamide/mesna in a 24-h i.v. continuous infusion appears to be tolerated and produces a biological response in ALL. We recommend that phase II studies of this drug schedule be conducted at the initial dose of 7 g/m2 repeated every 3 weeks and combined with other antileukemic agents. PMID- 9414392 TI - Expression of cyclin E in human gastric adenomas and adenocarcinomas: correlation with proliferative activity and p53 status. AB - The expression of cyclin E in human gastric adenomas and adenocarcinomas was examined immunohistochemically to elucidate the role of cyclin E in stomach carcinogenesis. The expression of cyclin E was detected in 49% (90/182) of the adenomas and 59% (260/439) of the adenocarcinomas. The incidence of strongly positive cases (overexpression of cyclin E) was significantly higher in the adenocarcinomas (29%; 128/439) than in the adenomas (4%; 8/182) (p < 0.01). The incidence of the cyclin E expression showed a tendency to be higher in deeply invasive carcinomas and in the cases with lymph node metastasis, while the incidence did not differ among histological types. The expression of cyclin E was significantly correlated with the proliferative activity of the tumor cells measured by KI-67 antigen expression (p < 0.01). It was also correlated with the abnormal accumulation of p53 protein in the tumor cells (p < 0.01). These results suggest that overexpression of cyclin E and subsequent deregulation of the cell cycle may confer the development and progression of the gastric carcinomas. PMID- 9414393 TI - Preclinical anticancer activity of cryptophycin-8. AB - Cryptophycin-8 was prepared by the conversion of the epoxide group on cryptophycin-1 to a chlorohydrin. In the studies reported here, cryptophycin-8 was evaluated for preclinical activity against subcutaneous tumors of both mouse and human origin. At the highest non-toxic single course treatment, the following results were obtained (Table A). Cryptophycin-8 was less potent than cryptophycin 1 by approximately 4-fold; however, it was both more water soluble and had greater therapeutic efficacy, as demonstrated by % T/C, tumor cell log kill values, range of dose effectiveness and host cures. PMID- 9414394 TI - MDL 201,307: a novel benzothiazepine modulator of multiple drug resistance. AB - A series of novel benzothiazepine derivatives were evaluated for their relative potential to reverse multiple drug resistance (MDR) phenotype in vitro as well as for their relative cardiovascular activity and neurotoxicity. Compounds were evaluated for antiMDR activity using Chinese hamster ovary cells with derived resistance to either vincristine or doxorubicin, or a human lymphoblastic leukemia line with resistance to vinblastine. Lead compounds with good antiMDR activity were further evaluated for their relative potential to exhibit cardiovascular and neurological pharmacodynamic activity. A single compound, MDL 201,307 with good antiMDR activity and low cardiovascular and neurologic activity was chosen for further study. In contrast to (R)-verapamil, MDL 201,307 showed only a weak potential to block calcium channels. Using a series of related murine fibrosarcoma cell lines (UV-2237M) with varying levels of resistance to doxorubicin, it was shown that MDL 201,307 augmented inhibition of growth due to doxorubicin. The antiMDR compound was also effective in enhancing the cytotoxicity of actinomycin-D and vinblastine although it was ineffective in increasing cytotoxicity of the nonMDR compound, 5FU. MDL 201,307 increased uptake and decreased efflux of doxorubicin suggesting that MDL 201,307 blocks the GP170 mediated efflux pump mechanism. MDL 201,307 represents a novel antiMDR agent with diminished potential for cardiovascular activity and neurologic interactions which presently limit many of the currently available first and second generations of antiMDR compounds. PMID- 9414395 TI - Comparative antitumor activities of halichondrins and vinblastine against human tumor xenografts. AB - Halichondrin B and homohalichondrin B are novel tubulin-interacting agents isolated from marine sponges. The in vivo antitumor activities of these compounds were examined in human tumor models in immunodeficient mice and rats. In nude mice, regression or pronounced delay of subcutaneous tumor growth was obtained with both halichondrins, at a maximum tolerable dose of 20 micrograms/kg Q2Dx5, in three of four vinblastine-sensitive tumors, including two melanomas and one osteosarcoma; one small-cell lung cancer line was resistant. The halichondrins as well as vinblastine showed only marginal activity against KM20L colon carcinoma xenografts. In a LOX melanoma lung colony formation assay in groups of six nude mice, all control animals were sacrificed because of respiratory symptoms 38 days after cell injection, and likewise one vinblastine-treated mouse after 53 days. All halichondrin-treated mice in the lung colony assay appeared healthy throughout an observation period of 112 days (p = 0.002). Upon necropsy all vinblastine-treated animals, and two of six mice in the halichondrin group, had macroscopic lung tumor colonies. In a nude rat model for LOX bone marrow metastases, the mean lifespan of untreated control animals was 15 days. Whereas vinblastine had only a marginal effect (17 days) in this model, halichondrin B prolonged the lifespan of the animals to 32 days, representing a significant (p = 0.0016) difference between the two compounds. In conclusion, the halichondrins, which comprise a subtype of tubulin-interactive anti-mitotic agents, showed distinct antitumor activity profiles in human tumor models, thereby encouraging their further preclinical development and possible clinical evaluation. PMID- 9414396 TI - Immunization of mice with irradiated melanoma tumor cells transfected to secrete lymphokines and coupled with IL-2 or GM-CSF therapy. AB - We compared the immunogenic activity of irradiated vaccines prepared from B16 F10 melanoma cells with one made from B16 F10 melanoma cells transfected with genes encoding murine IL-2 or GM-CSF. Vaccines were studied in the conditions of treatment of C57BL/6 mice with or without the corresponding lymphokines. Control and prevaccinated mice were challenged with parental B16 F10 murine melanoma cells (5 x 10(5)) subcutaneously in the midtail to examine growth of the primary (local) tumor in the middle of the tall and metastases to the lungs. This experimental model is very close to the clinical stages of metastatic melanoma. The effectiveness of preimmunization of mice was determined by the levels of antibody production to a melanoma-associated antigen termed B700. The comparison of antibody production, growth of primary melanoma tumors, number of mice surviving at the end of the observation period, mean survival time and per cent mice with metastases in the lungs showed that the best course of immunotherapy was prevaccination of mice with a vaccine of irradiated B16 F10 melanoma cells transfected to secrete GM-CSF, coupled with GM-CSF therapy. PMID- 9414397 TI - Prevention of mucositis in irradiated head and neck cancer patients. AB - Mucositis has been a significant debilitating side effect to the treatment of head and neck cancer by irradiation. A change in prophylactic treatment policy was made in August 1991 such that a total of 59 patients was divided into two groups. In Group A, 37 patients were treated prophylactically principally by sucralfate; while in Group B, 22 patients were treated by a three-part prophylactic regimen consisting of sucralfate, either cipriofloxacin or ampicillin, and clotrimazole troches. In Group A patients, unless they had limited fields as for stage 1 vocal cord patients, they uniformly progressed from patchy to confluent or severe mucositis (31/37 severe mucositis). Of the 22 patients in Group B, three patients also arrived at completion of treatment with severe or confluent mucositis; 12 of 22 experienced mild to patchy mucositis by completion, and seven of 22 retained their mucosa entirely intact at the completion of treatment. Four of 22 patients in Group B encountered a clinically diagnosed herpes virus infection of the oropharynx during treatment, which improved when Zovirax was added to their regimen. We conclude that mucositis is clearly and markedly improved by utilization of our prophylactic regimen for Gram negative rods and fungi. PMID- 9414398 TI - Tumoricidal potential of human macrophages grown in vitro from blood monocytes. AB - Adoptive transfer of activated autologous human macrophages obtained by in vitro differentiation of monocytes in culture has successfully undergone phase I clinical trials in patients with metastatic cancer. The efficacy of these autologous macrophages in the in vivo killing of human tumors has now to be demonstrated. GM-CSF was shown to increase the number of monocytes differentiating in culture into macrophages. These were then activated with IFN gamma which was reported as the best cytokine for tumoricidal activation of macrophages. The aim of this paper was to evaluate the in vitro tumoricidal activity of macrophages grown with GM-CSF and IFN gamma. This tumoricidal function was investigated by measurement of the cytolytic (chromium-51 release assay), cytostatic (anti-proliferative activity as measured by inhibition of [3H] thymidine incorporation in two different assays) and cytotoxic (MTT assay) activities on two tumor cells lines, U937 and K562 (respectively highly sensitive and resistant to soluble TNF alpha). Our results demonstrated that GM-CSF-grown macrophages exhibited significant cytolytic, cytotoxic and cytostatic activities on U937 cells. These were partly enhanced by IFN gamma activation. In contrast, they had no lytic and lower cytotoxic and cytostatic activities on K562 cells, and these were not modified by IFN gamma activation. Our data provide valuable information for future in vivo studies using adoptively transferred autologous macrophages. PMID- 9414400 TI - Differential chemosensitivity in oncogene-transformed cells. AB - The effects of anticancer drugs on cell growth have been investigated by MTT colorimetric assay using mouse and rat cultured cells transformed by various oncogenes and tumor viruses. Aclarubicin, mitomycin C and 1-hexylcarbamoyl-5 fluorouracil showed higher growth-inhibitory activities toward transformed cells than those toward the normal counterparts. Nimustine, bleomycin and 5 fluorouracil also showed selective growth-suppressive activities toward transformed cells except for a few cell lines. ras-oncogene-transformed cells were more sensitive toward 5-fluorouracil and 1-hexylcarbamoyl-5-fluorouracil than the normal parent cells and other transformed cells. These drugs would thus be effective in the chemotherapy of ras-induced cancer. PMID- 9414399 TI - Transcapillary forces of the subcutaneous tissue in patients treated with interleukin-2 and alpha-interferon: no capillary protein leak syndrome? AB - The purpose of this study was to evaluate whether treatment with interleukin-2 (IL-2) and alpha-interferon (IFN-alpha-2a) causes protein leakage from plasma to the interstitium, leading to the so called 'capillary leak syndrome'. This syndrome is supposed to cause dose-limiting side effects such as weight gain, edema and pleural effusions. Seven patients with disseminated malignant melanoma or renal carcinoma were studied before and after 5 and 12 days of treatment. Transcapillary forces were studied by measuring colloid osmotic pressures in plasma and interstitial fluid (on the thorax and ankle) with a 'wick' method. The colloid osmotic pressure in plasma was reduced by 30-35% during treatment, but with corresponding reductions in the interstitium. Hemoglobin, hematocrit, serum albumin and total protein decreased, whereas moderate edemas and weight gain were observed. These results demonstrate that during treatment with IL-2 and IFN-alpha 2a there are both fluid retention and augmented filtration of fluid from the vascular to the interstitial compartment, but no indication for a capillary leak syndrome. This may explain many of the cardiovascular side effects observed during such treatment. PMID- 9414401 TI - Phytohemagglutinin-induced gut hyperplasia and the growth of a mouse lymphosarcoma tumor. AB - NMRI mice injected subcutaneously with Krebs II lymphosarcoma cells and fed on a diet containing the kidney bean lectin phytohemagglutinin (PHA) within the range 0.45-7.0 mg/g diet, developed tumors during a 10 day period which on average were only 35% of the dry weight of tumors in lactalbumin (La) fed mice (control). The reduction in growth occurred in a dose-dependent manner in the range 0.45-3.5 mg/g diet. The degree of hyperplasia of the small intestine in response to feeding the PHA diets was higher in non-injected compared to injected mice. A lipolytic effect of PHA was observed above 1.75 mg/g diet in control mice and the highest concentration had a major effect on body weight. Since the index of hyperplasia at the lowest PHA concentration tested did not correlate with the reduction in tumor size, it is suggested that other factors in addition to the initial lectin-induced gut hyperplasia are involved in slowing down the progression of tumor growth. PMID- 9414402 TI - Comparison of triple helix formation by polypurine versus polypyrimidine oligodeoxynucleotides when conjugated to a DNA intercalator. AB - Biological applications of triplex forming oligonucleotides will require the development of oligomers with high avidity and specificity. We examined the binding enhancement resulting from intercalator conjugation to both parallel design (polythymidine T15) and antiparallel design (polypurine AG15, for binding a 15 base pair polypurine-polypyrimidine sequence in the IL-2R alpha gene enhancer) oligomers under various ionic strength and temperature conditions. Oligonucleotides were conjugated through a urea link to 6,9 diamino-3-methoxy acridine (to give T15C and AG15C). Intercalator conjugation dramatically enhanced the specific triplex binding avidity (Kd = 5 nM for AG15C and 275 nM for T15C at 25 degrees C, compared to 2 microM for AG15 and > 50 microM for T15 at 25 degrees C), without detectable binding to an inappropriate target sequence. Surprisingly, triplex formation with AG15C occurred at lower Mg2+ concentrations than with T15C. AG15 and AG15C showed rapid Mg2+ dependent self association, but not T15C or T15. T15C triplex formation occurred rapidly (completion in less than 4 min), while AG15C bound to its target sequence more slowly over 20-24 h. Thus, binding constants in the low nanomolar range are now achievable with intercalator conjugated polypurine antiparallel binding oligonucleotides, a prerequisite for biological applications of such agents. PMID- 9414403 TI - A phase II trial of paclitaxel in the treatment of recurrent or metastatic soft tissue sarcomas or bone sarcomas. AB - The study aimed to determine the activity and toxicity of taxol in the treatment of recurrent or metastatic soft tissue sarcomas or osteosarcomas. The major findings are that five patients had stable disease after two cycles of chemotherapy but two of these patients were subsequently removed from the study at their own request. The other three patients progressed after an additional two cycles of chemotherapy. Seven patients progressed during the first two cycles and were removed from the study. One patient completed only one cycle of therapy and was deemed inevaluable for study response. There were eight episodes of grade 3 or 4 neutropenia and two episodes of grade 3 thrombocytopenia. One patient experienced grade 3 neurological toxicity and one patient grade 3 mucositis. Two patients are currently alive with progressing disease and one patient is alive with no evidence of disease after undergoing surgery and radiotherapy. The principal conclusions are that Paclitaxel is ineffective in treating recurrent or metastatic soft tissue sarcoma and osteosarcoma. Treatment at this dose is quite myelosuppressive, but toxicity is generally manageable. Further study of this agent is not justified in this setting. PMID- 9414404 TI - Studies on the mechanism of action of 1-beta-D-arabinofuranosyl-5-azacytosine (fazarabine) in mammalian lymphoblasts. AB - Fazarabine has shown activity in the panel of 60 cultured human tumor lines of the National Cancer Institute. COMPARE analyses relating correlation coefficients of other anticancer drugs with those of fazarabine suggest that this agent operates through a similar mode of action to that of cytarabine. Studies have been carried out both in culture and in vivo to examine the mechanism of action of fazarabine in P388 murine and Molt-4 human lymphoblasts. Authentic fazarabine nucleotide standards were prepared by chemical and enzymatic methods and characterized on HPLC by comparison to related pyrimidine nucleoside-5' phosphates as well as by enzymatic digestion. Fazarabine inhibited the incorporation of labeled thymidine into DNA without influencing the synthesis of RNA or protein. Deoxycytidine overcomes this inhibition of DNA synthesis and also prevents the cytotoxicity of the drug to lymphoblasts, probably by competing for fazarabine uptake and metabolism. Fazarabine was rapidly phosphorylated in both cell lines; in P388 cells it was incorporated into DNA, where it continued to undergo the same type of ring opening and degradation as the free nucleoside. Alkaline elution studies demonstrated that exposure to the agent resulted in the formation of alkaline labile sites. Fazarabine also inhibited the methylation of deoxycytidine residues in DNA, but this effect was less pronounced than that produced by 5-azacytidine. Taken together, these studies suggest that fazarabine probably acts by arresting the synthesis and/or altering the structural integrity or functional competence of DNA. PMID- 9414405 TI - Chemopreventive efficacy of selenomethionine and its role in the antioxidant defense system in 2-acetylaminofluorene-induced hepatocarcinogenesis in rats. AB - Drinking water supplemented with selenium (8 ppm, daily) has been found to be highly effective in reducing tumor incidence and preneoplastic foci in 2 acetylaminofluorene (2-AAF) induced hepatocarcinogenesis in Sprague-Dawley male rats. Glutathione and several enzymes in liver tissue associated with antioxidant defense mechanisms, viz., catalase, glutathione-S-transferase, glutathione peroxidase, glutathione reductase and superoxide dismutase were investigated from hyperplastic nodules and non-nodular surrounding parenchyma. Treatment with selenomethionine either on initiation, or on selection/promotion, or during the entire experiment showed that selenom-ethionine was most effective when it was used as a supplement during the entire experiment, in terms of the antioxidant defense system and in reducing tumor incidence. Our results also confirm that selenium is particularly effective in limiting the action of 2-AAF during the initiation phase of hepatocarcinogenesis. PMID- 9414406 TI - Modulation of chemosensitivity in human colon carcinoma cells by downregulating protein kinase C alpha expression. AB - Protein kinase C (PKC) is thought to play a role in tumor progression and drug resistance of colon carcinomas. Specifically, the PKC alpha isoform has been implicated in drug resistance and responsiveness of colon carcinoma cells to growth factors. Therefore, in this study we determined the effect of downregulating PKC alpha expression by transfecting human colon carcinoma cells with an antisense PKC alpha expression vector and then determined the sensitivity of these cells to the anticancer drugs mitomycin C (MMC), 5-fluorouracil (5-FU) and vincristine (Vin). Transiently transfecting the human colon carcinoma cell lines Moser, SW480 and HT29 with antisense PKC alpha expression vector (but not antisense PKC beta expression vector) consistently increased the sensitivity of these cells to MMC, 5-FU and VIN by several-fold. Sensitivity to these drugs was then further determined in the Moser colon carcinoma cell line stably transfected with antisense PKC alpha expression vector. This stably transfected cell line, which expressed a high level of antisense PKC alpha RNA with a concurrent reduction of PKC alpha protein expression, was found to exhibit an increased sensitivity to these anticancer drugs. Thus, strategies designed to downregulate PKC alpha expression may have potential in improving the responses of colon carcinoma cells to cytotoxic drugs. PMID- 9414407 TI - Ultrastructural study of the urothelial lysosomal system in patients with transitional cell carcinoma after transurethral resection and interferon therapy. AB - The epithelial cells of human bladder urothelium contain a prominent lysosomal system on the surface layer which involves autophagic, phagocytotic and excretory processes. The noninvolved urothelium of tumor-bearing patients, however, does not contain this well-developed lysosomal system. Interferon restores the differentiation of the urothelium. Its action on the lysosomal system, however, has not been investigated. We studied ultrastructurally the noninvolved urothelium of eight patients with transitional cell carcinoma who after transurethral resection and intravesicular interferon instillations for 2 years did not develop recurrence. We noted that the number and size of lysosomes increased, being most numerous within the cells of the surface layer. Characteristic large lysosomes with the morphology of multivesicular bodies were also evident. These multivesicular bodies were almost entirely filled with small vesicles containing a dense core. Our findings show that after 2 years of interferon administration a re-appearance of a highly developed lysosomal system of the noninvolved urothelium was evident. This restoration to the normal morphology with reappearance of the lysosomal system, which could be partly attributed to interferon therapy, may be of clinical significance for prevention of tumor recurrence. PMID- 9414408 TI - Modulation of chemosensitivity by alpha interferon in multiple myeloma and non Hodgkin's lymphoma. AB - Low-grade non-Hodgkin's lymphoma and multiple myeloma are chemosensitive malignancies, but are rarely curable because of primary or acquired drug resistance. Interferon has been shown to modulate the multidrug resistance phenotype and to reinduce chemosensitivity in patients with chemoresistant tumors. Fifteen patients with multiple myeloma and 64 patients with low/intermediate grade non-Hodgkin's lymphoma unresponsive to initial chemotherapy were treated with alpha 2b interferon for 2 months. In case of an objective response, treatment was continued until disease progression; non responding patients received the same chemotherapy to which they were resistant, preceded by a 5 day course of interferon. Interferon salvage monotherapy induced an objective response in 1/15 patients with multiple myeloma and in 7/64 patients with non-Hodgkin's lymphoma. An objective response was achieved after retreatment with first-line chemotherapy preceded by interferon in 4/14 patients (28.6%) with multiple myeloma and in 20/56 evaluable patients (35.7%) with non-Hodgkin's lymphoma. Toxicity was moderate, predictable, manageable, and never caused interruption of the treatment. Interferon appears to be able to modulate chemosensitivity of tumors refractory to chemotherapy with several potential mechanisms, including an effect on drug accumulation; its utilization in this setting warrants further evaluation. PMID- 9414409 TI - The success of locoregional, low-dose recombinant interleukin-2 therapy in tumor bearing mice is dependent on the time of rIL-2 administration. AB - The extremely different administration schedules that are used in testing recombinant interleukin-2 (rIL-2) therapies for cancer may account for the extreme variation in efficacy reported in various studies in animal models. A major point may be the variation of the time interval between tumor transplantation and rIL-2 therapy. We hypothesized that administration of rIL-2 before the immune system has mounted a specific cellular reaction against the tumor (associated antigens) might result in lesser efficacies than later rIL-2 administration. This hypothesis was tested in DBA/2 mice bearing a syngeneic SL2 lymphoma. When 7000 IV/day rIL-2 was administered to tumor-bearing mice for 5 consecutive days starting on day 1, 3, 4, 5, or 6 after tumor inoculation, the survival curve of the mice did not significantly differ from that of diluent treated mice. In contrast, a significant difference was observed when treatment was begun on day 7, 8, 9, 10, or 12 (p < or = 0.004). rIL-2 therapies begun on day 9 or 10 were most effective, curing up to 80% of mice treated, despite there being an enormous burden of disseminated tumor present at that time (1-4% of the total body weight). When rIL-2 was administered for fewer than 5 consecutive days, beginning on day 10, the efficacy of the therapy dropped radically (p < or = 0.055). Involvement of a specific anti-tumor reaction was also tested. All mice that were cured of the tumor as a result of rIL-2 therapy proved to be specifically immune to the SL2 tumor. Furthermore, day 10-14 administration of rIL-2 was completely ineffective in CD4(+)-cell depleted mice (p = 0.0116 vs. rIL 2 therapy in non-depleted mice). Together, this implies that this form of rIL-2 therapy is mediated by tumor-specific T-cells. As a whole, these results indicate that T-cell mediated rIL-2 therapy of cancer in animal models is sensitive to the time when the rIL-2 is administered and to the length of time for which the rIL-2 is given. This should be taken into account when planning new therapy protocols and when analyzing published data. PMID- 9414410 TI - Neoadjuvant treatment of cancer of the bladder with M-VAC/M-VEC, and bladder sparing policy: preliminary report of a non-randomized study. AB - The life expectancy of patients with invasive bladder cancer is limited by the age of incidence and by the natural history of the cancer. Careful selection of patients, independent of age but linked to a neoadjuvant chemotherapy, should be useful for a bladder-sparing policy. Between January 1991 and December 1994, we selected 36 patients with invasive, transitional bladder cancer, but showing good performance status, after a transurethral resection biopsy performed with cytoreductive intention, and after a complete staging. Patients (median age, 65 years) were treated with neoadjuvant M-VAC/M-VEC and then selected for conservative surgery if the downstaging, topography, absence of in situ carcinoma, and residual bladder capacity allowed. At restaging, nine patients (27%) were in complete pathological response; 13 (39%) were in partial pathological response, with a total rate of 67%; and 11 patients (33%) were non responders, i.e. non-downstaged. Thirty nine percent were treated with radical cystectomy and 60% with limited surgery. Thirteen patients relapsed and seven died of disease during a median follow-up period of 23.5 months. At the end of the study, 68% of patients were alive, with a progression-free survival of 49.8% and a median survival of 32.9 months. Twenty one patients were alive at 31 December 1995, 14 with their bladder. No statistical differences were observed for overall survival and progression-free survival between the two surgery groups. Results were independent of age. A statistically significant difference was found (p = 0.0001) only between non-responders and all the downstaged patients, independent of surgery. These results confirm the feasibility of conservative treatment after a careful selection of patients, even in patients over 65 years, compared with standard available treatments. PMID- 9414412 TI - Metastatic potential and multidrug resistance correlation in the B16 melanoma system. AB - The question of whether metastatic potential and drug resistance are related phenotypes was addressed by comparing the biological behavior of the parental B16 melanoma and a multidrug resistant variant derived from it, the B16/Col/R. A more pronounced metastatic spread to lungs was observed in mice inoculated i.v. with the B16/Col/R variant than in those bearing the parental line. In addition, in the mice injected with the drug resistant melanoma, unusual tumor masses were observed. Large abdominal and spinal cord growths were seen with the MDR variant but not encountered in mice inoculated with the original B16 melanoma. We further attempted to test the capacity of the two cell types to perform several cellular functions relevant to the metastatic process. The B16/Col/R cells displayed a higher aggregability and cell motility than did the B16 cells. Adherence to endothelial cells was higher in the parental line than in the B16/Col/R, possibly supporting a more efficient extravasation of the variant cells. The drug resistant variant displayed a higher capacity to grow locally in kidney, spleen, cecum and peritoneum, as compared to the parental melanoma, indicating a higher ability of homing and growth in these potential target organs for metastasis. A correlation between metastatic potential and multidrug resistance appears therefore to exist in the system examined. PMID- 9414411 TI - Profile of T-cell receptor V beta gene usage of cytotoxic T cells induced by intrapleural administration of a streptococcal preparation, OK432, in malignant effusions. AB - T-cell receptor (TCR) gene rearrangements were analyzed in tumor-infiltrating lymphocytes (TIL) using the reverse transcription-polymerase chain reaction (RT PCR) to determine whether oligoclonal expression of TCRV beta occurs in TIL, and if so, whether it is involved in the clinical response and mechanisms of locoregional immunotherapy using a streptococcal preparation, OK432. Patients with malignant effusion of various origins were treated with intrapleural administration of OK432, and clinical responses were assessed by cytological and chest X-ray examinations. Pleural exudate cells (PEC), obtained before and after the administration of OK432 (designated as pre- and OK432-PEC, respectively), were subjected to TCR analysis. Both pre-PEC and OK432-PEC showed highly diverse expressions of TCRV beta gene usage in either type of PEC. The frequency of TCRV beta 20 gene expression in OK432-PEC was significantly higher than in pre-PEC. Moreover, the over-expression of the TCRV beta 20 gene usage was also induced in the peripheral blood lymphocytes and pre-PEC of patients by in vitro OK432 stimulation, but not in the PBL of one healthy volunteer. Single-strand conformational polymorphism (SSCP) analysis revealed the clonotypes of these TCRV beta 20 genes. Autologous tumor-specific killing activity could be detected in OK432-PEC and was significantly reduced by treatment with a TCRV beta 20-specific monoclonal antibody. These findings suggest that the rearrangement of TCRV beta 20 gene usage may be involved in the autologous tumor-specific action of malignant effusions in the treatment with OK432. PMID- 9414413 TI - Behavior of a novel monoclonal antibody for investigation of ovarian cancer and with possible involvement in multiple drug resistance. AB - Monoclonal antibodies (MAbs) were raised to an ovarian cancer cell line, OAW42, derived from a patient with a histology of serous cystadenocarcinoma of the ovary, in an attempt to identify novel antigens with a possible role in cancer medicine. One antibody P1H10, subclass IgG1, with a high titer was isolated and shown to recognize an antigen of 48 kDa. Enzyme-linked immunosorbent assay and immunocytochemical studies showed the presence of the target antigen in a number of carcinoma cell lines, including lung and breast, and in two out of three frozen breast tissue specimens. The antigen was not detected in normal human lymphocytes and there was minimal binding of the antibody to normal buccal cells. The antigen was not secreted by the OAW42 or the HepG2 cell lines and was not detected in the sera of a number of ovarian cancer patients. Indirect immunofluorescence studies confirmed the localization of the antigen to be intracellular. The binding of the antibody P1H10 to a number of multidrug resistant variants of the OAW42 cell line showed that the presence and the localization of the antigen in the drug-sensitive parental line and resistant variant cell lines was distinctly different and varied with the degree of drug resistance. The relative specificity of the antibody suggests it may be a possible diagnostic agent in human cancer. A possible role of the antigen in multiple drug resistance (MDR) is also suggested. PMID- 9414415 TI - Biochemical consequences of resistance to a recently discovered IMP dehydrogenase inhibitor, benzamide riboside, in human myelogenous leukemia K562 cells. AB - Benzamide riboside (BR) exhibits potent antitumor activity in a variety of cultured human tumor cells. The drug is metabolized to benzamide adenine dinucleotide (BAD), which in turn functions as a selective inhibitor of IMP dehydrogenase (IMPDH) activity with a Ki of 0.118 microM. In vitro, BR is a more potent antitumor inhibitor of IMPDH than tiazofurin, another IMPDH inhibitor which has shown significant oncolytic activity in adult patients with end-stage leukemia. To elucidate the mechanism of resistance, a variant of human myelogenous leukemia K562 cells was developed by subculturing sensitive cells in sublethal concentrations of BR over 60 generations. The BR resistant line that emerged exhibited an IC50 (a concentration producing 50% reduction in cell proliferation) of 148 microM, compared to the sensitive line which had an IC50 of 1.6 microM. The activity of the target enzyme, IMPDH, was increased 3-fold in the resistant variant. Studies on BR metabolism revealed that resistant cells formed only 18% of the active metabolite, BAD, compared to sensitive cells. This finding, in turn, correlated with the specific activity of NAD pyrophosphorylase (the enzyme responsible for the synthesis of BAD) which was reduced to undetectable levels in the resistant variant. The basal levels of NAD and guanylates were also significantly decreased to 41% and 48%, respectively, in the resistant line compared to the parent line. Additionally, after treatment with BR a decrease in guanylate level was observed only in the sensitive cells. Sensitive and resistant cells exhibit comparable cytotoxicity to agents outside the tiazofurin family, suggesting that a multidrug resistance was unlikely to explain the resistance to BR. Moreover, BR resistant cells exhibit collatoral sensitivity to 6-aminopurine, cytarabine and 5-fluorouracil, which have different mechanisms of action. In conclusion, these studies establish that the primary mechanism of resistance to BR involves an increase in IMPDH (target enzyme) activity with a concurrent decrease in NAD pyrophosphorylase (BAD synthetic enzyme) activity. PMID- 9414416 TI - In vitro evaluation of new anticancer drugs, exemplified by vinorelbine, using the fluorometric microculture cytotoxicity assay on human tumor cell lines and patient biopsy cells. AB - The feasibility of combined studies on a cell-line panel and primary cultures of patient tumor cells in the preclinical evaluation of new anticancer drugs was evaluated in a study of the activity and cross-resistance pattern in vitro of the new semi-synthetic vinca alkaloid vinorelbine (Vrb). The activity of Vrb was investigated in ten cell lines representing different resistance mechanisms and in a total of 256 fresh human tumor samples, using the fluorometric microculture cytotoxicity assay (FMCA). Resistance to Vrb in the cell lines was associated with expression of the multidrug resistance-mediating P-glycoprotein and the multidrug resistance-associated protein (MRP) and by a recently described tubulin associated mechanism, while the cell lines with topoisomerase II- and glutathion associated resistance did not show decreased sensitivity to the drug. Cross resistance to vincristine (Vcr) and other tubulin-active agents was high in cell lines as well as in patient cells. As with most commonly used anti-cancer drugs, Vrb was more active in hematological than in solid tumor samples. Among the solid tumors investigated, the highest in vitro response rates were observed in ovarian cancer (27%), sarcoma (25%), non-small cell lung cancer (21%) and bladder cancer (20%), while no response was observed in renal or colorectal cancer. Compared to Vcr, Vrb appeared to be slightly more active in solid tumors and slightly less active in hematological tumors. The results show that although Vrb displays a high degree of cross-resistance to Vcr and other tubulin-active drugs, some difference in the activity spectrum could be detected and that the drug is sensitive to multiple mechanisms of resistance. The results also suggest that leukemias, ovarian cancer, sarcoma and bladder cancer are possible further targets for Vrb. The combination of studies on a cell-line panel and patient tumor cells from a broad spectrum of diagnoses to evaluate a new drug seems feasible and may give information on the mechanism of action and target diagnoses for phase II trials. PMID- 9414414 TI - The effect of switching between a phytohemagglutinin-containing and a control diet on the growth and lipid content of a Krebs II lymphosarcoma tumor. AB - The present study concerns the importance of the timing of feeding mice a PHA containing diet (7 mg g-1 diet) on tumor formation. The major decrease in tumor weight occurred in mice fed on the PHA diet for 11 days. A marked reduction was also observed in animals pre-fed for 3 days with PHA before tumor cells were injected and the diet then changed to lactalbumin, La. A large decrease in tumor weight was also evident when a change of diet from La to PHA was made on the day of tumor cell inoculation. Despite the presence of the developing tumor PHA was able to induce hyperplasia of the small intestine in all groups of animals fed PHA during a part or the whole of the experiment. The dry weights of tumors attained in each of the experimental groups plotted as a function of duration of PHA feeding, and the percentage lipid content of the tumors, mirrored almost exactly one another, suggesting that the availability of essential lipid material is severely reduced by the lectin. This would appear to have a major effect on the observed reduction in tumor growth. PMID- 9414417 TI - Five-year survival of patients treated for esophageal cancer at the Institute of Oncology in Ljubljana in the years 1960-1989. AB - In last decades, the incidence of esophageal cancer in the male as well as female population of Slovenia has been increasing moderately. The number of long-term survivals is rather low. The treatment results of 13/714 (1.8%) 5-year survivors with esophageal cancer treated during the years 1960-1989 at the Institute of Oncology were analyzed. Different treatment modalities were used; the majority of patients were treated by using a combination of surgery, irradiation and chemotherapy. The analysis failed to establish a reliable cause of their significantly longer survival. Five years from the beginning of therapy 10/13 patients were alive without evidence of disease. Three patients developed a recurrence or another primary cancer. Longer survivals are probably attributable to a favorable treatment response associated with permanent or long-lasting disease free intervals and later recurrence. Longer and more frequent survivals can be expected with early stages and more effective, combined therapy for esophageal cancer. PMID- 9414418 TI - Hexadecylphosphocholine inhibits phosphatidylinositol and phosphatidylcholine phospholipase C in human leukemia cells. AB - Hexadecylphosphocholine (HePC) is the main representative of a new group of antineoplastic agents, the alkylphosphocholines. Besides remarkable antiproliferative properties on tumor cells in vitro and in vivo, HePC also induces differentiation and inhibits invasive growth of neoplastic cells. Knowledge of the molecular mechanisms by which HePC mediates its biological effects is poor. The observation that analogous substances, the alkyllysophospholipids, may interfere with lipid dependent intracellular signaling suggested similar mechanisms for HePC. We therefore investigated the effects of HePC on phospholipase C (PLC) activation in intact human leukemia cell lines. HePC inhibited fMLP induced phosphatidylinositol-specific PLC activation in HL60 cells and TNF-alpha induced activation of phosphatidylcholine-specific PLC in U937 cells. HePC reduced the number of TNF-alpha receptors on the surface of U937 cells by about 60%. Receptors for fMLP were not affected. Inhibition of TNF-alpha induced PC-PLC activation, however, seemed to be regulated at a post receptor level as PLC inhibition and receptor occupancy did not correlate. PMID- 9414419 TI - Percutaneous ablation of malignant liver tumor in rabbits using low radio frequency energy. AB - Radio frequency (RF) current has been used successfully to ablate normal human tissue. To further investigate the clinical application of this modality in tumors we studied the potential of using RF percutaneously to destroy experimental liver tumors. Thirty five outbred albino rabbits underwent liver VX2 tumor direct-implantation during open surgery. After 21 days ultrasonography was performed revealing tumor presence and size. A shielded RF needle was designed so that it could be inserted percutaneously through an introducing needle, and an electrical insulation shield covering the RF needle could be retracted to control the length of the exposed RF needle inside the tissue. Twenty two days after tumor implantation RF was applied via the aforementioned needle using a ZoMed International RF generator. In one group of rabbits the procedure was performed under direct vision during open surgery and on the other group treatment was applied percutaneously, guiding the needle by tumor palpation. Rabbits were killed 3 days later and pathology revealed 4 to 25 mm intratumoral RF induced lesions. A direct relation was found between lesion size, power and duration of RF application (At 7.5 W, r = 0.48, p = 0.032). Based on our preliminary results we may conclude that RF may have clinical application in the near future for percutaneous local tumor control in parenchymal organs. PMID- 9414422 TI - The association of the H-ras oncogene and steroid hormone receptors in gynecological cancer. AB - Expression of the ras family of cellular oncogenes is associated with tumorigenicity, invasiveness and metastatic potential in a variety of human carcinomas. Additionally, H-ras cooperates with glucocorticoids and with ovarian hormones in cell transformation and in the development of mammary carcinomas. Steroids are considered to be tumor promoters and their levels influence the cure rates and survival of the patients with gynecological lesions. It is proposed that they exert tumor promoting activity by transcriptional regulation of nuclear proto-oncogenes, such as c-fos, c-jun, and c-myc. The human H-ras gene contains within its first and fourth introns, sequences that are specifically recognized by glucocorticoid and estrogen receptors, respectively. Using gel retardation assays, the level of steroid receptor binding in H-ras elements has been compared, employing nuclear extracts from human endometrial and ovarian lesions and from the adjacent normal tissue. Elevated binding of the glucocorticoid and estrogen receptors in the corresponding H-ras elements in almost all tissue pairs tested has been found. It is suggested that the H-ras proto-oncogene is hormonally regulated and directly implicated in human gynecological cancer through elevated, steroid-induced gene expression. PMID- 9414420 TI - Ki-67 and p53 immunoreactive stain and early colorectal neoplasms. AB - The aim of this work was to conduct a histopathological study of the different pathways of colorectal tumorigenesis by using Ki-67 and p53 immunoreactive stains. Between 1987 and 1993, 14,023 patients were investigated by colonoscopy at Akita Red Cross Hospital and several affiliated hospitals. Five hundred and sixty-five cases of early colorectal neoplasms identified in this population were used in the present study. Specimens were cut in half: one half was cut along the vertical axis, the other half along the horizontal axis to analyze the structure of glands which opened to the surface. They were stained with hematoxylin-eosin, Ki-67 and p53 immunoreactive stains. macroscopically, early colorectal neoplasms were classified into two types: protruding type and depressed type. The protruding type was mainly composed of branching glands. In contrast, the depressed type was almost completely composed of straight glands which opened to the surface. The Ki-67 positive ratio was almost equal. However, the p53 positive ratio was significantly different in the protruding type and the depressed type. The results suggest the existence of at least two pathways of colorectal tumorigenesis: one with the process of branching structure and one without the process of branching structure. Furthermore, there were clinical and immunohistochemical indications that the depressed type invades into deeper layers more rapidly than does the protruding type. PMID- 9414421 TI - Establishment of human prostate tumor xenografts in nude mice and response to 9 nitrocamptothecin in vivo and in vitro does not correlate with the expression of various apoptosis-regulating proteins. AB - Flow cytometry and microscopy analyses have demonstrated that 9-nitrocamptothecin (9NC) induces apoptosis in prostate carcinoma LNCaP, DU-145 and PC-3 cells grown in culture or as xenografts. 9NC induces apoptosis regardless of the ability of the cells to induce tumors following xenografting into nude mice. Detection of apoptosis by flow cytometry was preceded or accompanied by increased cell size, loss of nuclear structure and vacuolization, as the tumor regressed, but no visible chromatin fragmentation. This is the first demonstration that 9NC is curative for human prostate carcinoma xenografts in the nude mouse model in the absence of detectable drug-induced toxicity during and after tumor regression. These findings indicate that 9NC may develop into a chemotherapeutic drug for the effective treatment of prostate cancer patients. Further, there was no apparent correlation of the steady-state level of the apoptosis-regulating proteins, Bcl 2, Bcl-XL, Bax and Ich-1, with tumorigenicity of the prostate cells xenografted in nude mice, aggressiveness of tumors grown in nude mice, and induction of apoptosis by 9NC. However, the TIAR protein was present at markedly high levels in all prostate carcinoma cell lines and this may correlate with their susceptibility to 9NC-induced apoptosis. PMID- 9414423 TI - Stimulation of GALT and activation of mesenteric lymph node lymphocytes by a modified lysozyme in CBA mice with MCa mammary carcinoma. AB - Lysozyme (hen egg-white lysozyme) and its derivative mPEG-lyso (lysozyme coupled with polyoxyethyleneglycol) were tested in CBA mice bearing MCa mammary carcinoma for their effects on intestinal mucosal immunity (GALT) and mesenteric lymph node lymphocytes (MLNL), after oral administration. Following a cycle of administration of 100 mg/kg/day lysozyme or 350 mg/kg/day mPEG-lyso for 9 consecutive days, GALT was analyzed by using optical histology, and mesenteric lymph node lymphocytes were studied by cytofluorimetric analysis of CD3, CD4 and CD8 antigens, and of DNA and RNA content following in vitro culture with concanavalin A. Both lysozymes significantly increase the number of lymphatic nodules on gut epithelium as determined by histological analysis of sections of small bowel. mPEG-lyso, unlike native lysozyme, gives protection from the decline of the blastogenic activity of MLNL observed at early stages of tumor growth, as shown by the increased nucleic acid content of these cells. On the same cells, both lysozyme and mPEG-lyso also seem to prevent the decline of CD4+ cells observed during tumor growth in control animals. These data confirm the effects of lysozyme on GALT and show that the new lysozyme derivative mPEG-lyso has effects on host immunity greater than those of the native molecule. PMID- 9414424 TI - Hematological modifications induced by estrogens in cirrhotic animals. AB - Estrogens are known to have mitogenic activities and to influence hematic crasis. A wide literature on this subject allows us to re-evaluate the results of research performed in 1950. Estrogens injected into the peritoneum (estradiol propionate 2.5-5 mg, twice weekly) or into tibial bone marrow (estradiol propionate 1.5 mg weekly) of guinea pigs, after carbon tetrachloride induced cirrhosis (CCI4 inhalation for 7-15 min twice weekly), caused hyperchromic anemia, with erythroblastosis and leucocytosis; hyperplasia of reticulo hystiocitary cells of bone marrow and lymphoid organs; splenic hemosiderosis; hyperplasia of immature bone marrow cells, with maturative inhibition; and terminal bone marrow hypoplasia. Our results agree with recent literature data concerning the role played by natural estrogens and environmental xenoestrogens that are capable of stimulating bone marrow progenitor cell proliferation and delaying their maturation, by the secretion of growth factors, especially from hyperplastic stromal cells, in solid and hematological tumors. PMID- 9414425 TI - Lymphocyte number and stress parameter modifications in untreated breast cancer patients with depressive mood and previous life stress. AB - Depressive mood disorders and severe, chronic stressful life events (DSM-III-R criteria) were more frequently diagnosed in 106 breast cancer patients with respect to 37 patients with benign breast diseases (control group) (p < 0.001), during a stressful period such as hospital admission, diagnosis uncertainty, and when awaiting surgery. The study was performed 5 +/- 3 days before histological diagnosis had been done. Controls showed reduced 24-h diuresis and low catecholamine excretion (norepinephrine, NE; and epinephrine, E) that positively correlated with 24-h diuresis (p < 0.001) and CD3+ lymphocytes (p = 0.056), as during a normal stress response. In contrast, breast cancer patients showed increased 24-h diuresis (with respect to controls p < 0.001) and catecholamine values (p < 0.05). Patients' 24-h diuresis correlated positively with NE (p = 0.02) and 17-ketosteroids (p = 0.004); blood cortisol correlated positively with CD3+ (p = 0.01), CD4+ (p = 0.02), CD8+ (p < 0.01), CD16+ (p = 0.01) lymphocytes and negatively with E (p < 0.03); catecholamines correlated negatively with CD8+ (p = 0.006). These preliminary data are discussed in relation to upregulation of the adrenergic system and the different mechanisms of immune system regulation involved in breast cancer patients, compared with those in subjects with benign breast disease. The differences in these mechanisms may be a result of an imbalance of the bi-directional regulatory circuit of the psycho-neuro-endocrine immune system, caused by previous life stress or the presence of the tumor mass. PMID- 9414428 TI - Induction of apoptosis in an androgen-independent mouse cell line by transforming growth factor-beta 1. AB - Androgen-dependent tumors eventually progress to androgen-independent tumors after androgen withdrawal. Androgen-independent CS2 cells could grow in serum free culture whether androgen is present in the medium or not. In the present study, the mechanism of cell death in CS2 cells was examined after transforming growth factor-beta 1 (TGF-beta 1) treatment. Based upon the temporal sequence of DNA fragmentation, morphologic changes and loss of cell viability, these cells underwent apoptosis with TGF-beta 1 treatment. During the apoptotic process induced by TGF-beta 1, mRNA expression of testosterone repressed prostatic message-2, glucose regulated 78 kDa protein, and calmodulin increased. Flow cytometric analysis showed that TGF-beta 1 treatment resulted in a block in the G0/G1 phase of the cell cycle. These results demonstrate that although androgen withdrawal could not induce apoptosis in androgen-independent CS2 cells, these cells retain the ability to undergo apoptosis induced by TGF-beta 1. This system should be a good model for investigating apoptosis of hormone-independent cancer. PMID- 9414427 TI - Induction of resistance in the human leukemia cell line HL60 towards hexadecylphosphocholine and other ether phospholipid analogues. AB - Hexadecylphosphocholine (HePC) is a new ether lipid analogue with remarkable antineoplastic activity in vitro and in vivo. As the precise molecular mechanism by which this substance and probably other ether lipids exert their biological effects is still not defined, we tried to approach this problem by generating a cell line resistant to the antiproliferative properties of HePC. This was successfully accomplished by slow adaptation over a period of 14 months, of the very sensitive human leukemia cell line HL60. HePC resistant HL60 cells (HL60R) tolerate 8- to 10-fold higher doses of HePC and are continuously cultured in medium containing 10 micrograms/ml of HePC. An immunophenotypic and karyotypic characterization of HL60 and HL60R cells showed only marginal differences between the two cell lines. Total phospholipids, total cholesterol, protein and vinyl ether lipid content were equal in both cells. A down-regulation of the ether lipid mass in HL60R of about 40% could reflect one mechanism of tolerance induction. Though HePC uptake in HL60R cells was significantly lower than in the parental line, steady state measurements of cellular HePC content revealed similar HePC content in the membranes at HePC concentrations that were cytotoxic for HL60 but did not affect HL60R. This observation indicates that uptake and cellular accumulation of HePC do not determine HePC resistance. The resistant HL60R cells also showed a considerable degree of cross-resistance to ether phospholipids ET-18-OCH3 and BM 41.440, suggesting a common mode of action for HePC and other ether lipid analogues. PMID- 9414426 TI - Quantitative measurements of the efficacy of new anti-cancer agents on fresh human AML cells by using multivariate flow analysis. AB - The testing of new human leukemia-specific drugs for activity against primary acute myeloid leukemia (AML) blasts is severely limited by the low and variable clonogenic potential of primary human leukemias in culture. To circumvent this problem, we have modified a previously described flow cytometric approach to permit the simultaneous determination of live/dead cells, and the quantitation of the surviving cell fraction as a ratio of viable cells in treatment and control groups. The method utilizes the combination of calceinAM as a probe for intracellular esterase activity (green fluorescence,) which has the advantage over carboxyFDA of pH insensitivity and superior signal-to-noise ratio, and propidium iodide (red fluorescence) as an indicator of plasma membrane integrity. Suspension cultures of AML blood and marrow samples from patients were treated with known active agents as well as several new agents arising from a clonogenic disk assay screen. Quantitative dose-response values obtained from surviving cell fractions assayed by flow cytometry at 24 h following drug exposure demonstrated the utility of this assay for quantitating drug-induced cytotoxic effects on primary human AML cells in short-term culture. PMID- 9414430 TI - The effect of TNP-470 on cell proliferation and urokinase-type plasminogen activator and its inhibitor in human lung cancer cell lines. AB - The plasminogen activator system has been found to modulate neoplastic spread and angiogenesis in tumors. An angiogenesis inhibitor, TNP-470, has been shown to possess potent antitumor activities in various types of cancer cells. We therefore investigated the inhibitory effect of TNP-470 on cancer cell proliferation, and the suppressive effect of TNP-470 on urokinase-type plasminogen activator (u-PA) and its inhibitor (PAI-1), in human lung cancer cell lines in vitro. TNP-470 inhibited cell proliferation in a dose-dependent manner in both cell lines. u-PA and PAI-1 in culture supernatants were suppressed with the concentrations of TNP-470, in association with a decrease in viable cancer cells. Unchanged serum levels of u-PA and PAI-1 would be of great advantage to the diseased patients, since the plasminogen activator system has a crucial function in the process of distant metastasis. PMID- 9414429 TI - Regression of C6 rat brain tumors by cells expressing an antisense insulin-like growth factor I receptor RNA. AB - We have previously shown that C6 cells expressing an antisense insulin-like growth factor I receptor (IGF-IR) RNA are no longer tumorigenic in syngeneic rats, protecting them from subsequent subcutaneous tumor challenge and causing regression of established subcutaneous tumors. In the present study, we have investigated the efficacy of this strategy on intracerebrally implanted C6 rat glioblastoma cells. We demonstrate that C6 cells expressing an antisense IGF-IR RNA implanted for 24 h in the subcutaneous tissue of the rats are able to elicit an anti-tumor response in the brain, leading to complete brain tumor regression and long-term survival of the rats. These findings suggest the possibility of therapeutic intervention in human gliomas. PMID- 9414431 TI - Prolactin signal transduction mechanisms in the mammary gland: the role of the Jak/Stat pathway. AB - Prolactin signal transduction in mammary epithelial cells is mediated by a novel, direct signalling system that links the activation of the prolactin receptor at the cell surface to changes in gene transcription in the nucleus. This recently identified pathway is a variant of the Jak/Stat (for Janus kinase/signal transducer and activator of transcription) pathway used by many other growth factors and cytokines. Current data suggest that the key intracellular components of the prolactin signalling pathway are the kinase Jak2 and the transcription factor Stat5. This discovery has exciting implications for the interaction between prolactin and other extracellular signals in both the mammary gland and other tissues. Here we review work that began with attempts to understand the regulation of milk protein gene expression and ultimately demonstrated the central role of the Jak/Stat pathway in prolactin signal transduction in the mammary gland. PMID- 9414433 TI - Oxytocin: cellular and molecular approaches in medicine and research. AB - In May 1995 the third Hanseatic Endocrine Conference at Stade, Germany, attracted 140 scientists from all over the world to summarize the current knowledge on one hormone--oxytocin. This article presents the major findings of the meeting with the realisation that oxytocin provides major model systems with which to elaborate a whole series of novel endocrinological paradigms, as well as being the example of choice for establishing revolutionary new techniques, which will no doubt spread to studies of other hormone systems. The papers from this symposium will be published in full. PMID- 9414434 TI - Control and regulation of folliculogenesis--a symposium in perspective. AB - The Intervet Symposium 'Control and Regulation of Folliculogenesis' took place at the Annual Conference of the Society for the Study of Fertility in University College, Dublin in July, 1995. Five papers were presented, of which abstracts have been published. Discussion centred on monotocous species and principally addressed the question 'How is the ovulatory follicle selected and nurtured?' Many follicles start to develop simultaneously, but most become atretic. During the luteal phase of the cycle, several waves of follicles are initiated, and although one in each wave gains dominance, all ultimately atrophy. The dominant follicle in the wave produced towards the end of the luteal phase ovulates. The biochemical characteristics of dominant and subordinate follicles were contrasted, and the endocrine environment in which they waxed and waned was analysed. Interaction of autocrine and paracrine effectors, principally with FSH, LH and their receptors, ultimately determines follicular destiny. PMID- 9414432 TI - Glutathione as a treatment for male infertility. AB - The excessive generation of reactive oxygen species (ROS) by abnormal spermatozoa and by contaminating leukocytes has been identified as one of the few defined aetiologies for male infertility. As a consequence, work has begun on evaluating the role of antioxidants in the management of these patients. Glutathione plays a significant role in the antioxidant defences of the spermatogenic epithelium, the epididymis, and perhaps in ejaculated spermatozoa. The use of antioxidants in vitro appears to be of value in preserving fertilizing capacity, although no clinical data are available. Glutathione administered in vivo to patients who may have infertility secondary to excessive oxidative stress appears to act at the epididymis and during spermatogenesis, to improve the function of ejaculated spermatozoa. However, fertility studies have not yet been conducted. Controlled studies of glutathione and other antioxidants in patients with defined ROS pathology are urgently required. PMID- 9414435 TI - Identification of early pregnancy factor as chaperonin 10: implications for understanding its role. AB - Early pregnancy factor (EPF) is a secreted substance with growth regulatory and immunomodulatory properties. It is required for successful establishment of pregnancy and for proliferation of both normal and neoplastic cells, in vivo and in vitro. The rosette inhibition test was used as a bioassay, and the appearance of EPF in serum in the very early stages of pregnancy (in mice, within 4-6 h of mating) was first described two decades ago. However, because of the difficulty of this bioassay and the paucity of EPF in biological materials, the primary structure of the molecule has been identified only recently. Seventy per cent of the amino acid sequence of EPF derived from human platelets was determined. With the exception of a single residue, this was identical to the sequence of rat mitochondrial chaperonin 10 (cpn10). Cpn10 is a heat shock protein that functions as a molecular chaperone. It binds to and stabilizes cpn60 and, in concert, these molecules mediate protein folding in mitochondria, chloroplasts and bacteria. Characterizing EPF as an extracellular form of cpn10 raises unprecedented questions about the mechanism of action. It may be that, as a molecular chaperone in the extracellular compartment, EPF can functionally modify other proteins, serving as a regulator of regulators. PMID- 9414436 TI - Soluble sperm factors and Ca2+ release in eggs at fertilization. AB - All eggs are activated at fertilization by an increase in intracellular free Ca2+. How the spermatozoa triggers the release of intracellular Ca2+ has not been established in any species. One hypothesis is that spermatozoa introduce a Ca(2+) releasing factor into the cytoplasm after gamete membrane fusion. It has been suggested that spermatozoa may introduce Ca2+ itself, or a small molecular weight factor that triggers the release of Ca2+ from intracellular stores. However, these suggestions are not consistent with a number of experiments in vertebrate eggs. Here, I present the argument that spermatozoa cause intracellular Ca2+ oscillations in mammalian eggs by introducing a specific protein called an oscillogen. It is suggested that it does not mediate its effects by causing increased inositol, 1,4,5-trisphosphate (InsP3) production, but instead directly affects the ability of the stores to undergo Ca(2+)-induced Ca2+ release. The sperm oscillogen hypothesis may help explain the patterns of intracellular Ca2+ waves that occur during fertilization in hamster eggs, as well as the ability of an injection of live spermatozoa to activate development in human eggs. PMID- 9414437 TI - Cell cycle co-ordination in embryo cloning by nuclear transfer. AB - Exciting new opportunities in embryo cloning have been made possible by recent studies on the interaction of the donor nucleus with the recipient cytoplasm after embryo reconstruction. This article reviews information regarding the co ordination of nuclear and cytoplasmic events during embryo reconstruction, in particular the direct and indirect effects of maturation/ meiosis/mitosis promoting factor (MPF), upon the transferred nucleus. This will be discussed in relation to DNA replication, the maintenance of correct ploidy, the occurrence of chromosomal abnormalities and development of reconstructed embryos. Although this review is primarily concerned with the reconstruction of mammalian embryos, specific examples from amphibians will also be cited. PMID- 9414438 TI - Nutrition, insulin and polycystic ovary syndrome. AB - The adverse effects of obesity on reproductive function in women are well recognized, but women with polycystic ovary syndrome (PCOS), the most common cause of anovulatory infertility, seem particularly vulnerable to the effects of excessive intake of calories. Polycystic ovary syndrome is associated with hyperinsulinaemia and insulin resistance, the causes of which remain unclear. These metabolic abnormalities are, in turn, related to a disorder of energy expenditure, characterized by reduced post-prandial thermogenesis. It is proposed that these closely interlinked phenomena that, particularly in overweight subjects, are associated with anovulation, may confer a biological advantage for women with PCOS at times of food deprivation, when such women may reproduce more successfully than those without PCOS. A possible causal link between hyperinsulinaemia and ovulation is explored by reference to the interaction of insulin and LH in granulosa cells. PMID- 9414440 TI - Nitric oxide in the human uterus. AB - Nitric oxide (NO) plays a crucial role in many biological systems. Recent evidence indicates that NO is found in the human uterus. During pregnancy, it is produced in the placenta, the decidua and the myometrium. In the nonpregnant state, nitric oxide synthase, the enzyme that catalyses the production of NO, has been identified in both the myometrium and the endometrium. Potential roles for NO in the human uterus are speculative, but include vasodilatation (both before implantation, and in the uteroplacental and systemic circulation during pregnancy), inhibition of platelet activation during menstruation, and suppression of myometrial contractility during pregnancy. Nitric oxide may also be involved in uterine pathology. Excessive NO production by the uterus during menstruation could lead to menorrhagia. During pregnancy, a change in NO production may be implicated in pre-eclampsia, and animal studies have shown that inhibition of NO production leads to intrauterine growth retardation. If a role for NO is confirmed in these various uterine conditions, pharmacological modification of NO activity may lead to novel therapeutic applications. However, these notions are still conjectural, and extensive work is required before such treatments can be introduced into clinical practice. PMID- 9414439 TI - In vitro maturation of mammalian spermatozoa. AB - During epididymal transit, mammalian spermatozoa undergo maturation and acquire full fertilizing capacity. The contribution of factors from the epididymal epithelium appears to be essential for this process. Although complete in vitro maturation of epididymal spermatozoa has not been achieved, stages of maturation can be induced under various conditions. The most successful results have been obtained by incubating epididymal spermatozoa with primary cultures of epididymal epithelium. These co-incubation methods promote sperm motility and the capacity of spermatozoa to bind to and fertilize oocytes, and extend the viability of spermatozoa in vitro. Specific androgen-dependent secretory proteins from epididymal principal cells that may be involved in this maturation process have been identified using pulse-labelling techniques. PMID- 9414441 TI - Oxytocin: a paracrine regulator of prostatic function. AB - It is now well established that the peptide oxytocin can act as a paracrine factor as well as a classic hormone. Oxytocin is produced locally in both the testis and ovary, where it may modulate both steroidogenesis and contractility of the male and female reproductive tracts. The peptide is also present in the prostate and seminal fluid and there is growing evidence that oxytocin may be produced in the prostate. Within the prostate, oxytocin has been shown to increase growth of the epithelial tissue and increase both muscular tone and contractile activity. Furthermore, prostatic concentrations of the peptide are regulated by androgens. It is hypothesized that oxytocin may act as a paracrine factor to regulate cell growth and that this may be secondary to its effects on the enzyme 5 alpha-reductase which converts testosterone to dihydrotestosterone. In addition, oxytocin may be involved in the pathophysiology of benign prostatic hyperplasia. PMID- 9414442 TI - Genetic conflict in early development: parental imprinting in normal and abnormal growth. AB - Parental (genomic) imprinting is the process by which the differential expression of maternal and paternal alleles at certain genetic loci in mammalian embryos occurs. Such loci are implicated in the control of fetal, placental and neonatal growth, and, more generally, in diverse aspects of fetal nutrient acquisition and maternal-fetal interactions. Not surprisingly, the aberrant expression of imprinted genes is implicated in a range of embryonic and fetal abnormalities. We outline how an evolutionary theory, based on classic parent-offspring conflict theory, relates to certain fetal growth abnormalities. In particular, we suggest that growth abnormalities resulting from the manipulation of preimplantation mammalian embryos in vitro (for example large calf syndrome) may reflect the occurrence of genetic conflict over the fetal growth programme in the early preimplantation period. PMID- 9414443 TI - Neoteny and the thyroid in ratites. AB - The ratites (for example ostriches, emus) are neotenous descendants of flying birds. The best studied cases of neoteny among vertebrates are in the Amphibia. In this class, whether individuals metamorphose and breed as adults, or whether they become sexually mature as neotenous aquatic larvae, is controlled by the thyroid. In this review it is argued that the thyroid may have been important in the evolution of ratite neoteny. The evidence is based on characteristics of ratites that could indicate thyroid abnormality, similarities between ratites and thyroidectomized non-ratite birds, and preliminary results from a study of thyroid function in ostriches. PMID- 9414444 TI - Protein phosphorylation/dephosphorylation in the meiotic cell cycle of mammalian oocytes. AB - The meiotic cell cycle in mammalian oocytes commences in the fetal ovary, proceeds up to the diplotene stage of the first prophase, and is arrested around birth at a G2-like phase. Reinitiation of meiosis, which occurs immediately before ovulation, represents the transition from G2 to M phase of the cell cycle. Resumption of meiosis is associated with a reduction in cAMP concentrations within the oocyte and is negatively regulated by an activated cAMP-dependent protein kinase (PKA). These findings led to the hypothesis that meiotic arrest is mediated by a PKA-phosphoprotein substrate, which undergoes dephosphorylation under conditions of decreasing intra-oocyte concentrations of cAMP. Thus far, a phosphoprotein that serves as a substrate for PKA and maintains meiotic arrest has not been identified. Nevertheless, the idea that regulation of enzyme activities of a series of protein kinases and phosphatases governs the progression through the meiotic cell cycle is now commonly accepted. The present knowledge of the specific phosphorylation/dephosphorylation biochemical events that participate in the control of meiosis in mammalian oocytes is discussed in this review. PMID- 9414445 TI - Inflammatory mediators and parturition. AB - A role for inflammatory mechanisms in ovulation and parturition has been proposed on many occasions. In addition, many agents directly implicated in parturition, such as cytokines and prostaglandins, can be pro-inflammatory. The cervix can be softened and labour induced in women by mechanical trauma (sweeping the membranes), prostaglandins, antiprogestins or chemokines such as interleukin 8 (IL-8). This review presents evidence that invading cells, such as neutrophils, may be involved in the softening of the cervix and the onset of birth, and describes pathways in which this action can be controlled by cytokines and vasodilatory agents such as prostaglandins. Such mechanisms may, in turn, be controlled by steroids. The role of progesterone is enigmatic since progesterone concentrations do not fall in the peripheral circulation of women at the time of birth and yet antiprogestins soften the cervix in a manner indistinguishable from that seen during parturition. Prostaglandin E (PGE) can be both pro-inflammatory and anti-inflammatory, the former action probably occurs at the level of the blood vessel, whereas the latter is associated with the change in cytokine profile induced by PGE. This latter effect may contribute to pregnancy maintenance by maintaining a favourable cytokine profile in decidua. In contrast, the pro-inflammatory action of PGE may be involved in a synergistic action with chemokines such as IL-8 and play a role in parturition. In early pregnancy decidua, this pro-inflammatory action is likely to be controlled by progesterone dependent prostaglandin dehydrogenase associated with the small blood vessels in decidua. PMID- 9414446 TI - Transgenic animals and gonadotrophins. AB - A variety of transgenic animal models has been developed to elucidate in vivo the functions of gonadotrophin genes. Some of these have focused on regulatory aspects through expression of gonadotrophin subunit promoter-driven reporter genes. Others have been carried out by overexpression or targeted disruption of specific gonadotrophin subunit genes, or by eliminating pituitary gonadotroph cells in transgenic mice by expressing toxic transgenes under gonadotrophin subunit promoters. In addition, the overexpression or knock-out of genes of other hormones (for example GH and the inhibin subunits) has elucidated the regulation of gonadotrophin gene expression. Many of the transgenic animals produced serve as good models for human diseases affecting the hypothalamic-pituitary-gonadal function. This review summarizes the key results obtained with these novel genome modification techniques on the physiology and pathophysiology of gonadotrophin synthesis and secretion. PMID- 9414447 TI - The vascular endothelium in normal pregnancy and pre-eclampsia. AB - Hypertensive disorders of pregnancy affect up to 15% of pregnancies and the most severe form, pre-eclampsia, is the leading cause of maternal death in the UK today. Pre-eclampsia is a multisystem disorder affecting virtually every organ and system in the body, with hypertension and proteinuria, the traditional diagnostic features, representing two facets of a complex pathophysiological process. The common pathological feature of the disease, whether in the decidual vessels of the placental bed, renal microvasculature, liver, heart or cerebral circulation, is vascular endothelial damage and dysfunction. Before new ways of preventing or ameliorating hypertensive disorders of pregnancy can be found, we must first understand the pathophysiological mechanisms underlying the clinical problem. This review summarizes the evidence that endothelial dysfunction plays a key role in hypertensive disorders of pregnancy and will focus on the most severe form, pre-eclampsia. PMID- 9414448 TI - Regulation of reproductive hormone secretion in primates by short-term changes in nutrition. AB - In primates, as in nonprimates, it is well established that periods of chronic or severe undernutrition can lead to a suppression of reproductive hormone secretion. Recent studies in monkeys and humans indicate that reproductive hormone secretion begins to be suppressed with even very brief periods of undernutrition. Specifically, in male monkeys reproductive hormone secretion responds to missing a single meal, eating a single meal, and to a change in the timing of daily meal intake. These findings suggest that nutritional/metabolic signals that are linked to food intake are part of the normal physiological mechanism regulating the daily activity of the reproductive axis, rather than simply signals that influence the activity of the reproductive axis in pathological conditions of severe undernutrition. The nature of the metabolic cue(s) linking reproductive hormone secretion to subtle changes in the metabolic status of the body remains unknown, as does the route by which metabolic information reaches the central hypothalamic neurones governing reproductive hormone secretion. However, recent studies indicate that the metabolic cue transmitting information to the reproductive axis is dependent on calorie intake, but is not dependent on changes in body mass or composition, changes in intake of a specific nutrient, changes in plasma glucose or insulin concentrations, or signals emanating from the taste or smell of food or the physical process of food ingestion. PMID- 9414449 TI - Control of meiotic arrest. AB - Oocytes of many species arrest at specific cell cycle stages during their development. An external signal from a hormone or the fertilizing sperm causes them to resume the cell cycle. The control of meiotic arrest can be usefully formulated in terms of the interaction between cell signalling mechanisms and the protein machinery that controls the cell cycle. Much of what we know about cell messengers, particularly calcium, and the cell cycle control proteins comes from work on oocytes. Recent work on cell signalling pathways in mammalian cells and cell cycle control in yeast has been essential to our understanding of meiotic arrest in oocytes. PMID- 9414450 TI - Ca2+ release and the development of Ca2+ release mechanisms during oocyte maturation: a prelude to fertilization. AB - Oogenesis involves the production of an oocyte that can undergo fertilization and support early development. The stimulus that initiates embryogenesis is an increase in the concentration of intracellular Ca2+ in the cytoplasm of the oocyte at the time of fertilization. The development of the ability of the oocyte to release Ca2+ in response to the fertilizing spermatozoon is an essential step in the process of oogenesis. Mammalian oocytes are particularly useful for studying the development of Ca2+ signalling systems, owing to the series of Ca2+ oscillations generated at fertilization, compared with the monotonic Ca2+ increase seen in nonmammalian species. Recent evidence has revealed that Ca2+ release mechanisms are modified during oogenesis. The maximal sensitivity of Ca2+ release is reached in the final stages of oocyte maturation, just before the optimal time for fertilization. In this review, we consider the mechanism underlying Ca2+ release in mammalian oocytes and discuss how the release mechanisms are modified during oocyte maturation. The tight co-ordination of the differentiation of the Ca2+ signalling system with the development of the oocyte provides a means of ensuring successful activation at the time of fertilization. Finally, we consider the consequences for embryo development in circumstances in which the co-ordination is lost. PMID- 9414451 TI - Roles of mesenchymal-epithelial interactions and hepatocyte growth factor-scatter factor (HGF-SF) in placental development. AB - The major components of the mammalian placental membranes are an epithelial surface layer, the trophoblast, and a heavily vascularized mesenchyme, the allantoic mesenchyme. The trophoblast layer makes the most intimate contact with maternal tissues and it displays a wide range of unusual, often invasive, phenotypes. However, one common feature of trophoblast development in all species is a strong correlation between the proliferation and differentiation of this epithelial layer and its physical contact with developing allantoic mesenchyme. This suggests an epithelial-mesenchymal interaction involving paracrine signals from allantoic mesenchyme acting on adjacent trophoblast. The expression patterns of several growth factors and their receptors, including hepatocyte growth factor scatter factor (HGF-SF) and its receptor, c-met, support the hypothesis. Furthermore, HGF-SF and c-met gene knockout studies in mice indicate that HGF-SF and c-met are both essential for placental development. HGF-SF, in addition to being a potent mitogen, causes scattering and morphogenic changes in cultured cells and is believed to be an important mediator of the induction of epithelial differentiation during embryogenesis. This review evaluates the importance of mesenchymal induction of trophoblast growth and differentiation in placental development and argues that HGF-SF is a crucial component of the mesenchymal stimulus. PMID- 9414452 TI - Fertilization of oocytes by injecting spermatozoa, spermatids and spermatocytes. AB - The feasibility of fertilization by injecting spermatozoa into oocytes has increased significantly the possibilities for treatment of severe male infertility. However, the rapidity of human application has raised some concern about potential health hazards for the progeny. Human pregnancies and births have also occurred with the use of immature spermatozoa and spermatids, and normal offspring have been born after the injection of secondary spermatocytes into mouse oocytes. This short review deals with the problems that may arise from the injection technique and from the use of deficient or immature sperm cells for fertilization, with particular attention to human applications. Tests for screening parents and follow-up of children are suggested to control the main suspected risk factors. PMID- 9414453 TI - The pars tuberalis of the pituitary: a gateway for neuroendocrine output. AB - The pars tuberalis is a structurally distinct region of the adenohypophysis, the function of which has been unclear for decades. Recent studies, which demonstrate the localization of melatonin receptors on the pars tuberalis, suggest a photoperiodic function. The principal cell type of the pars tuberalis is morphologically distinct from others in the pituitary and is thought to secrete a specific product. In support of this, evidence is emerging that ovine pars tuberalis cells secrete a factor ('tuberalin') that exerts hormonal control over both gene expression and prolactin release from the pars distalis lactotrophs. These data in conjunction with physiological studies, which show that photoperiodically driven cycles in prolactin secretion can occur in the absence of an intact hypothalamic-pituitary axis, suggest that the function of the pars tuberalis is to act as an endocrine intermediate in the photoperiodic effects of melatonin on prolactin secretion. Studies of the cellular biochemistry of the ovine pars tuberalis suggest that the main function of melatonin is to prevent or terminate transcriptional and translational activation by an unidentified factor (Stim X). On the basis of these physiological and biochemical studies, a hypothetical model is proposed to account for the mechanism of photoperiodic regulation of prolactin secretion by melatonin. PMID- 9414454 TI - Apoptosis and ovarian function. AB - For decades, the mechanisms responsible for germ cell depletion from the ovary, either directly during the perinatal period or indirectly via follicular atresia during postnatal life, have remained relatively obscure. The recent application of sensitive biochemical techniques for the study of cell death to the analysis of ovarian function has revealed that these two events, as well as a third instance of ovarian cell degeneration (luteolysis), are dependent upon the activation of physiological cell death mechanisms. It is therefore hypothesized that the controlled deletion of ovarian cell populations is accomplished via activation of a 'universal' pathway of cellular suicide involving altered expression of a conserved cohort of genes. The identity of the hormonal and intracellular effectors responsible for the coordination of life and death decisions made by ovarian cells during development as well as the biological and clinical implications of gene-directed cell death in the ovary are explored in this review. PMID- 9414455 TI - Mechanisms mediating the response of GnRH neurones to excitatory amino acids. AB - Excitatory amino acids, such as glutamate, exert a profound stimulatory effect on the reproductive axis of several mammals. Although glutamate receptor agonists stimulate GnRH secretion, both in vivo and in vitro, it is unclear whether GnRH neurones respond directly to glutamatergic excitation. Immortalized GnRH neurones (GT1 cells) express glutamate receptors when grown in culture and also show enhanced GnRH secretion in response to glutamate receptor agonists. In addition, immunocytochemical evidence at the electron microscope level supports the possibility of a direct interaction between glutamatergic and GnRH neurones. In general, however, double-label histochemical studies (using immunocytochemistry, in situ hybridization, or a combination of these techniques) have not shown significant glutamate receptor gene expression in GnRH neurones of adult animals. It remains to be determined whether a higher degree of glutamate receptor gene expression occurs during development. This general lack, or very low amount, of glutamate receptor gene expression in the GnRH neurones of adults supports the view that excitatory amino acids exert their stimulatory action on the reproductive axis primarily through interneuronal pathways that impinge on the GnRH neurones, rather than by stimulating GnRH release directly. PMID- 9414457 TI - Intracellular mechanisms triggering gonadotrophin secretion. AB - The recently cloned GnRH receptor, a G-protein coupled receptor that spans the membrane seven times, plays a central role in the maintenance of normal reproductive events. In pituitary gonadotrophs, activation of the GnRH receptor stimulates a concert of intracellular signalling pathways. Phospholipase C stimulation generates inositol 1,4,5 trisphosphate and diacylglycerol, which release calcium and activate protein kinase C, respectively. After these primary signals, prolonged activation of protein kinase C arises from the continued production of diacylglycerol from additional signal transduction pathways. While characteristic calcium responses, involving specific calcium pools, are instrumental in triggering exocytosis, the precise role of protein kinase C activation is unclear. Key players within the exocytotic machinery are also elusive but may include a range of membrane, guanine nucleotide and calcium binding proteins, inositol 1,4,5 trisphosphate receptors and the cell cytoskeleton. Cellular signalling is also important in determining pituitary responsiveness to GnRH, involving intracellular cross-talk between the GnRH, oestradiol and progesterone receptors. PMID- 9414456 TI - Angiogenesis in the ovary. AB - In adult tissues, capillary growth (angiogenesis) occurs normally during tissue repair, such as in the healing of wounds and fractures. Inappropriate capillary growth is associated with various pathological conditions, including tumour growth, retinopathies, haemangiomas, fibroses and rheumatoid arthritis in the case of rampant capillary growth, and nonhealing wounds and fractures in the case of inadequate capillary growth. The female reproductive organs exhibit marked, periodic growth and regression, accompanied by equally striking changes in their rates of blood flow. It is not surprising, therefore, that they are some of the few adult tissues in which angiogenesis occurs as a normal process. Ovarian follicles and corpora lutea have been shown to contain and produce angiogenic factors. These angiogenic factors appear to be heparin-binding and to belong to the fibroblast growth factor (FGF) and vascular endothelial growth factor (VEGF) families of proteins. In addition, factors regulating gap junctional communication may play a critical role in coordinating the interactions between luteal vascular and nonvascular tissues. Further elucidation of the specific physiological roles of these factors in follicular and luteal growth, development and function will ultimately lead to improved methods for regulating fertility in mammals. PMID- 9414458 TI - Transgenic mice in the analysis of reproductive development and function. AB - Transgenic mice have become important model systems for studying molecular, cellular, organ, and whole animal physiology. In particular, transgenic technology allows determination of cell lineage differentiation and the role(s) of specific proteins in mammalian development and oncogenesis. Transgenic mice can be created that express wild-type genes, mutant genes, marker genes or cell lethal genes in a tissue-specific manner. In addition, homologous recombination strategies in embryonic stem cells permit more sophisticated manipulation of the mammalian genome, including the functional deletion of specific genes in whole mice or in a specific tissue. Since all transgenic mice created must be bred to study the consequences of transgene or mutant allele expression, a number of effects of these genome manipulations on the reproductive development and function of these mice have been uncovered. In this review, we summarize the transgenic mouse models in which defects and abnormalities in the reproductive axis have been demonstrated. PMID- 9414459 TI - Noradrenergic regulation of cyclic GnRH secretion. AB - The GnRH cells represent the final output neurones of an integrated neuronal network used by the brain to generate pulsatile LH secretion from the pituitary gland. Changes in LH secretion profile throughout the ovarian cycle, including the preovulatory LH surge, result principally from alterations in the output of this GnRH network and it has been a key goal of many neurobiologists to elucidate the components and nature of this network. This review documents recent progress in understanding the role of noradrenaline within the GnRH network and highlights and explains its 'enabling' or permissive characteristics. Network behaviour analysis suggests that noradrenaline should be considered as a permissive agent promoting high output states of the GnRH network. On the basis of recent molecular and neuroanatomical data, it is proposed that oestrogen influences brainstem noradrenergic neurones specifically within the nucleus tractus solitarius to facilitate synaptic transmission within the GnRH network. In this manner, noradrenaline is likely to play a role in bringing about the increased GnRH messenger RNA expression and secretion necessary for ovulation. PMID- 9414460 TI - HLA-G and pregnancy. AB - Recent studies of the nonclassical HLA-G class I gene provide insight into its function(s) during pregnancy. The HLA-G gene can be transcribed in different isoforms resulting from alternative splicings and encoding membrane-bound and soluble proteins. These different mRNA species have been found in the various trophoblast cell subpopulations that constitute the maternofetal interface in the human placenta. The raising of antibodies to HLA-G has introduced new tools to determine in which types of trophoblast cells and in which other tissues these transcriptional isoforms are translated in functional proteins. The HLA-G gene exhibits a certain amount of polymorphism, the exon three that encodes the alpha 2 external domain showing the most extensive nucleotide variability. It remains to be determined whether the homozygosity of some HLA-G alleles constitutes a real disadvantage in terms of pregnancy or resistance to specific pathogens. Regarding the potential antigen-presenting function(s) of HLA-G, two isoforms are capable of binding an identical set of nonamer peptides derived from a variety of intracellular proteins. The ligand motif contains three anchor residues and is similar to that of classical HLA class I molecules. Experiments are being performed to identify the recognizing cells and to determine whether HLA-G induces a cytolytic (including anti-viral) T-cell response or in some other way represses natural killer-cell functions. PMID- 9414461 TI - Prolactin receptor subtypes: a possible mode of tissue specific regulation of prolactin function. AB - Prolactin mediates its effect on target cells through an interaction with membrane-anchored receptors. In the last decade, several subtypes of the receptor have been isolated from different species. This has generated a great deal of interest in the roles of the receptor subtypes and the possible divergent signalling pathways in mediating the pleiotropic effects of prolactin on target tissues. Our current knowledge of the signalling pathway of prolactin is derived mainly from the interaction of the hormone with one of its receptor subtypes (the long form) isolated from rats. In vitro expression studies have led to the identification of the regions within the long form prolactin receptor that are essential for the association of the tyrosine kinase Jak-2, and the phosphorylation events leading to activation of the prolactin responsive beta casein promoter. To date, a specific target gene that may be activated after interaction of prolactin with the short form of the receptor has not been identified. However, the different receptor subtypes are present in the same cell type in vivo and their expression is hormone regulated, possibly through multiple promoters that control transcription of the prolactin receptor gene. Comparative studies suggest that the signalling pathways and the relevance of different receptor subtypes on prolactin function may vary between species. PMID- 9414462 TI - The human sperm centrosome is responsible for normal syngamy and early embryonic development. AB - As early as 1887, it was postulated that the mature oocyte possesses all of the elements necessary for embryonic development with the exception of an active division centre, and that the spermatozoon contains such a centre, but lacks the substrate in which to operate. This division centre is called the centrosome. The precise definition of this structure is still a subject for debate. It consists of two centrioles in a perpendicular arrangement and pericentriolar material, and is considered to be responsible for nucleation of microtubules and the formation of the mitotic spindle. There is a paternal pattern of inheritance of the centrosome in humans; thus, human oocytes lack centrioles but the spermatozoa carry two. At gamete fusion the sperm tail is incorporated into the ooplasm, and the centriolar region forms the sperm aster while the sperm head is decondensing; this aster acts to guide the female pronucleus towards the male pronucleus. The centriole duplicates during the pronuclear stage, and at syngamy centrioles are found at opposite poles of the first cleavage. The centrosome has several implications for human infertility. It is possible that immotile or nonprogressively motile spermatozoa may possess centriolar abnormalities or an absence of centrioles. Similarly, antisperm antibodies against centrioles may be responsible for mitotic arrest. One way of solving this problem would be the use of donor centrosomes. To this end, we have assessed the ability of embryos injected with physically separated sperm segments (head only, head and tail separated or isolated tail) to develop normally. Fluorescent in situ hybridization revealed an almost universal mosaicism in these embryos, suggesting that physical disruption of the spermatozoa compromises the ability of the centrosome to function in the zygote. Thus far, centrosome donation with centriole-carrier flagellae obtained by this dissection method does not appear to be feasible. PMID- 9414463 TI - Central oxytocin and reproductive behaviours. AB - Oxytocin is a neurohypophyseal hormone that has long been associated with uterine contraction during parturition and milk ejection during nursing. Recent studies have suggested that oxytocin is also a neurotransmitter that has central effects important for reproduction, including the initiation of parental and sexual behaviours. This review describes oxytocin pathways in the brain and examines their regulation by gonadal steroids. Brain oxytocin receptors are remarkable for their plasticity and for striking species differences in their distribution. The molecular characterization of this receptor has provided several clues to the regulation of its expression. Comparative and transgenic studies suggest that central oxytocin release may influence reproductive behaviours, but the importance of these central effects depends on the pattern of expression of the receptor--a pattern that is species-specific. PMID- 9414464 TI - Testicular leukocytes: what are they doing? AB - Leukocytes, specifically macrophages, lymphocytes and mast cells, are found within the testes of most, if not all, mammals. In some species (for example, rats, mice and humans), the number of 'resident' testicular macrophages, in particular, is quite considerable. However, reproductive biologists are only beginning to explore the characteristics and possible biological significance of these cells. As in other tissues, the testicular leukocytes are involved in immunological surveillance, immunoregulation and tissue remodelling. They are implicated in the mechanisms that make the testis a particularly successful site for tissue transplantation in some experimental animals. Moreover, recent studies have demonstrated that the testicular macrophages have specific trophic effects on Leydig cell development and steroidogenesis. In turn, the development and functions of the testicular leukocyte population are clearly influenced by the testicular environment, and especially by the Leydig cells and Sertoli cells. These data indicate an important role for leukocytes in testicular homeostasis. Balanced against this beneficial role is the fact that these cells possess the potential to damage testicular function in conditions of immune activation, as their inflammatory and cytotoxic activities may disrupt the normal environment of the testis. The importance of the testicular leukocytes to normal and abnormal testicular function is evident. The challenge for future research is to define the details of this relationship. PMID- 9414465 TI - Reactive oxygen species and sperm physiology. AB - Although high concentrations of reactive oxygen species (ROS) cause sperm pathology (ATP depletion leading to insufficient axonemal phosphorylation, lipid peroxidation and loss of motility and viability), recent evidence demonstrates that low and controlled concentrations of these ROS play an important role in sperm physiology. Reactive oxygen species, such as the superoxide anion, hydrogen peroxide and nitric oxide, induce sperm hyperactivation, capacitation or the acrosome reaction in vitro. The ROS involved in these processes may vary depending on experimental conditions, but all the evidence converges to describe these events as 'oxidative' or 'redox regulated'. Human sperm capacitation and acrosome reaction are associated with extracellular production of a superoxide anion that is thought to originate from a membrane 'oxidase'. The enzymes responsible for tyrosine phosphorylation-dephosphorylation of sperm proteins are possible targets for ROS since mild oxidative conditions cause increases in protein tyrosine phosphorylation and acrosome reaction. The lipid peroxidation resulting from low concentrations of ROS promotes binding to the zona pellucida and may trigger the release of unesterified fatty acids from the sperm plasma membrane. The fine balance between ROS production and scavenging, as well as the right timing and site for ROS production are of paramount importance for acquisition of fertilizing ability. PMID- 9414466 TI - Neuroendocrine regulation of gonadotrophin II release and gonadal growth in the goldfish, Carassius auratus. AB - The goldfish, a member of the carp family, is a widely used model for reproductive neuroendocrine studies of economically important fish. The two gonadotrophin (GTH) molecules released from the fish anterior pituitary, GTH-I and GTH-II, are structurally similar to tetrapod FSH and LH, respectively. Gonadotrophin II is the best studied, and in goldfish stimulates gonadal growth and steroidogenesis, ovulation and sperm release. Growth hormone also has gonadotrophic actions in fish which enhance gonadal steroidogenesis. The principal stimulatory and inhibitory systems regulating GTH-II release are the gonadotrophin-releasing hormone (GnRH) and dopamine neurones in the preoptic hypothalamic region. In goldfish there are two native GnRH forms, salmon GnRH and chicken GnRH-I; both stimulate GTH-II release but use different signal transduction pathways. In contrast to mammals, teleost fish do not have a median eminence and the GTH-II cells are thus directly innervated by neurones producing GnRH, dopamine and other stimulatory neurohormones. For most of these factors, the ability to stimulate GTH-II release varies seasonally. The amino acid neurotransmitter, gamma-aminobutyric acid, has the most prominent stimulatory actions which enhance GnRH release and inhibit dopamine turnover in the hypothalamo-pituitary complex. Neuropeptide Y stimulates GTH-II release by a combined direct action on the gonadotroph and also by enhancing GnRH release. Positive and negative sex steroid feedback mechanisms act concurrently to regulate GTH-II release in adults of both sexes. The principal site of positive feedback is the GTH-II cell where testosterone and oestradiol potentiate GnRH stimulated GTH-II release. Negative feedback by sex steroids involves activation of inhibitory dopamine neurones, thus maintaining tight control over circulating GTH-II concentrations. PMID- 9414467 TI - Environmental oestrogens--present understanding. AB - Three years ago it was hypothesized that the reported adverse changes in male reproductive health could be explained by exposure to compounds with oestrogenic (or other hormone disruptive) activity. Although this issue has been highly publicized, there has been little progress towards a realistic assessment of whether environmental oestrogens pose a health risk to humans. This article attempts to give a brief overview of the current status of knowledge concerning environmental oestrogens and highlights some of the difficulties associated with risk assessment. Compounds within several major groups of chemicals, including organochlorine pesticides, polychlorinated biphenyls, phenolic compounds and phthalate esters, have been identified as being weakly oestrogenic by in vitro and in vivo screening methods. Many of these compounds are widespread and persistent in the environment. They are likely to be present in the food chain, drinking water, plastics, household products and food packaging, although which is the most important route of human exposure is unclear. In addition to exposure to man-made chemicals, the consumption of plant-derived oestrogens in foodstuffs poses a potential risk to human health as phytoestrogens are more potent oestrogens and their intake by some infants is likely to be considerable. PMID- 9414468 TI - Nuclear transfer and reprogramming. AB - Nuclear transfer techniques for mammalian embryos have been developed in the last decade. Embryonic nuclei from advanced stages of preimplantation development can be fully reprogrammed and the totipotency is restored when nuclei are transferred into ooplasts. Transfer of nuclei after gene expression from the embryonic genome has started does not appear to restrict the reprogramming of these nuclei. The principles of nuclear transfer are outlined with respect to nuclear remodelling, nucleocytoplasmic interactions and effects of the cell cycle. However, the molecular mechanisms involved in reprogramming donor nuclei remain unknown. It is proposed that epigenetic DNA modification, such as DNA methylation that regulates gene expression, is related to the reprogramming of transplanted nuclei. PMID- 9414469 TI - Roles of reactive oxygen species in the regulation of luteal function. AB - Ephemerality and prolongation of luteal function have been matters of great concern in reproduction for many years. However, their control mechanisms are very complex and differ among mammals. Recently, evidence has indicated that reactive oxygen species may play important roles in the regulation of luteal function. Reactive oxygen species are present in most somatic cells and are involved in apoptosis, or 'physiological cell death'. In the corpus luteum, reactive oxygen species also exert luteolytic effects as well as some paradoxical luteotrophic effects. This paper discusses the possible roles of reactive oxygen species in the control of luteal function. PMID- 9414470 TI - Adhesion molecules in implantation. AB - At implantation, trophectoderm attaches to the apical uterine luminal epithelial cell surface. Molecular anatomy studies in humans and mice, and data from experimental models have identified several adhesion molecules that could take part in this process: integrins of the alpha v family, trophinin, CD44, cad-11, the H type I and Lewis y oligosaccharides and heparan sulfate. The endometrial cell surface mucin MUC1 may play a role in both steric inhibition of attachment and selective glycan display. After attachment, interstitial trophoblast invasion occurs requiring a new repertoire of adhesive interactions with maternal extracellular matrix as well as stromal and vascular cell populations. Human anchorage sites contain columns of cytotrophoblasts in which self-attachment gives way progressively to adhesion to extracellular matrix and then interstitial migration. The beta 1 integrins are important during these later stages of implantation and placentation. PMID- 9414471 TI - In vitro culture of ovarian follicles. AB - This review offers a practically oriented introduction to follicle culture in vitro, focusing on mouse follicles, but with reference to other species. The main principles of follicle growth are addressed, including the constraints of tissue culture, methods of follicle isolation, and techniques for individual and collective culture of intact follicles. Culture systems that support a spherical or a non-spherical follicular structure in vitro are discussed in terms of follicular and oocyte development, and methods for assessing follicular function in vitro are presented. Oocyte development in most in vitro culture systems is currently suboptimal and the parallel development of oocytes and follicles is discussed, with a view to maintaining the competence of the oocyte. Finally, some potential future applications of follicle growth in vitro are suggested. PMID- 9414472 TI - Fetal undernutrition and disease in later life. AB - Recent findings suggest that coronary heart disease and stroke, and the associated conditions, hypertension and non-insulin dependent diabetes, originate through impaired growth and development during fetal life and infancy. These diseases may be consequences of 'programming', whereby a stimulus or insult at a critical, sensitive period of early life results in long-term changes in physiology or metabolism. Animal studies provide many examples of programming, which occurs because the systems and organs of the body mature during periods of rapid growth in fetal life and infancy. There are critical windows of time during which maturation must be achieved; and failure of maturation is largely irrecoverable. PMID- 9414473 TI - Advances in understanding gonadotrophin-releasing hormone receptor structure and ligand interactions. AB - Gonadotrophin-releasing hormone (GnRH) is the central regulator of the reproductive system and its analogues are used widely in the treatment of diverse diseases. The GnRH receptor is a member of the large family of G-protein-coupled receptors (GPCRs) which have seven transmembrane domains. Knowledge of these receptors has assisted the development of molecular models of the GnRH receptor that allow prediction of its three-dimensional configuration and the way GnRH binds and activates its receptor. Comparison with other GPCRs led to the discovery that Lys121, in the third transmembrane domain, has a role in agonist binding. The history of GnRH structure-activity studies has allowed the identification of an acidic residue in the third extracellular loop of the receptor that is required for binding of mammalian GnRH, while synthetic GnRH analogues have showed that Asn102, in the second extracellular loop, may interact with the carboxy-terminus of GnRH. These residues can now be incorporated into the receptor models that are being used to design orally active non-peptide GnRH analogues for contraception and treatment of a variety of reproductive disorders. PMID- 9414474 TI - Preimplantation growth factor physiology. AB - The robust independence of preimplantation embryo development in vitro suggests that the developmental programme is autonomous. The rapid accumulation of evidence during the last decade for participation of many hormones, growth factors and their receptors in these early stages of embryogenesis has challenged this conclusion. In this review, the insulin and epidermal growth factor families, which have been best studied in mice, are used to illustrate the different roles growth factors may play in preimplantation physiology and the circuits that possibly mediate their participation. Tumour necrosis factor alpha, an inhibitory factor and growth hormone previously considered to be restricted to orchestrating postnatal growth and development, is also discussed. In the absence of results indicating the existence of a master regulatory factor, the data support the hypothesis that the redundancy of growth factor actions may provide fail-safe protection to the preimplantation developmental programme. PMID- 9414475 TI - Placental 11 beta-hydroxysteroid dehydrogenase: barrier to maternal glucocorticoids. AB - During mammalian pregnancy, the circulating concentration of cortisol (in rodents, corticosterone) in the mother is much higher than that in the fetus. Since the placenta is the only barrier, apart from the uterus, between the mother and her fetus, this gradient in cortisol concentrations suggests that there is a placental barrier preventing maternal cortisol from crossing into the fetus. The intracellular enzyme 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD) is an ideal candidate for this barrier because it interconverts cortisol and corticosterone to their inactive metabolites cortisone and 11 dehydrocorticosterone. Indeed, 11 beta-HSD enzyme is expressed in the placenta of humans and a range of other animal species. Moreover, it is well positioned to serve as the barrier since it is localized to the syncytiotrophoblast, the site of maternal-fetal exchange. Given that fetal exposure to excessive amounts of glucocorticoids leads to intrauterine growth retardation, it has been hypothesized that the physiological significance of this placental 11 beta-HSD barrier is to protect the fetus from adverse effects of maternal glucocorticoids. PMID- 9414476 TI - Biology of the relaxin-like factor (RLF). AB - The relaxin-like factor (RLF) is a novel member of the insulin-IGF-relaxin family of hormones and growth factors. Also known as the Leydig cell insulin-like factor (Ley-I-L), this peptide and the mRNA encoding it appear to be expressed in very large quantities in the Leydig cells of the testis. However, it is also produced in the ovary of a number of species in both follicular theca cells and in the corpus luteum of the cycle and pregnancy. RLF gene transcripts have been identified at a much lower level of expression throughout the bovine female reproductive tract and also in the hypothalamus. Although data are limited, it would appear that RLF represents a new differentiation-related factor with specific functions linked to reproductive physiology in male and female mammals. PMID- 9414477 TI - Ovarian follicular dominance: the role of intraovarian growth factors and novel proteins. AB - Folliculogenesis is associated with the development of a group of follicles at various stages of maturation from which a species-specific number of follicles are selected for continued growth. These selected follicles, after being exposed to the requisite hormonal environment, ovulate in response to the preovulatory gonadotrophin surge. Follicular dominance is the mechanism by which the selected follicle(s) undergoes rapid development in an environment where growth and development of other follicles, recruited at a similar time, are suppressed. These processes are controlled by the interaction of endocrine signals and locally produced ovarian growth factors. The response of the two major follicular cell types, granulosa and theca cells, to gonadotrophins is regulated by the local production of growth factors. Mechanisms controlling growth factor action occupy a central role in the regulation of folliculogenesis. In this review, we highlight the influence of the extracellular matrix in this process by describing its involvement in regulating the activity of components of the insulin-like growth factor system, transforming growth factor beta superfamily, fibroblast growth factors and the epidermal growth factor/transforming growth factor alpha family. In addition, some recent studies on the role of protein factors produced by the dominant follicle in maintaining dominance and inhibiting the growth of subordinate follicles are described. PMID- 9414478 TI - Three-dimensional structure of the zona pellucida. AB - The zona pellucida is the extracellular coat that surrounds the mammalian oocyte. It forms a spherical shell of remarkably uniform thickness (5-10 microns in eutherian mammals). The mouse is currently the largest source of data on the zona pellucida and this review is built largely on these data. The zona pellucida is composed of three proteins in both mice and humans: ZP1, ZP2 and ZP3. These proteins are glycosylated and, in mice, have mature relative molecular masses of 200,000, 120,000 and 83,000, respectively. ZP1 is a dimer of two apparently identical subunits. All three mouse proteins have been sequenced and possess transmembrane domains at their C-terminal ends coupled with furin cleavage sites immediately upstream. Sequence data have been used to provide an accurate assessment of the mole ratios of the three proteins. The ratio of ZP2:ZP3 is close to 1:1, whereas ZP1 is approximately 9% of the combined mole amounts of ZP2 and ZP3. Ultrastructural evidence suggests that the mouse zona pellucida is composed of filaments constructed by head-to-tail association of globular proteins. The coordinate synthesis of the three zona pellucida proteins coupled with the near 1:1 stoichiometry of ZP2 and ZP3 is consistent with a model in which ZP2-ZP3 heterodimers are the basic repeating units of the filament, with cross-linking of filaments by dimeric ZP1. This model is also consistent with data from ZP2 and ZP3 gene knockout and antisense experiments. However, the structure remains unproven. The small amount of ZP1 relative to ZP2 and ZP3 may have important implications for the distribution of ZP1 cross-links, since the number of cross-linking sites potentially exceeds the number of ZP1 dimer molecules by a considerable margin. The evidence that ZP1, ZP2 and ZP3 are all synthesized via a membrane-bound step is discussed and two models are proposed for the assembly of the zona pellucida. The cortical reaction and its effect on the zona pellucida are examined in detail. It is shown that the amount of material released by cortical granules could be of the order of 30% by mass of ZP1, and that if this material was distributed predominantly on the inner face of the zona pellucida, its local concentration could approach that of ZP1. A model in which the zona block to polyspermy is caused by direct titration of zona pellucida binding sites is suggested as an alternative to the explanation that relies on enzyme cleavage of ZP2 to ZP2f. Finally, some of the major experimental and structural issues that remain to be addressed are identified. PMID- 9414479 TI - Models for male infertility: the t haplotypes. AB - The t haplotypes are variant alleles of genes in the proximal region of mouse Chromosome 17, linked together by four inversions. While females carrying two t haplotypes are fertile, males are sterile. Their spermatozoa exhibit severe motility defects and are unable to penetrate zona pellucida-free oocytes. Spermatozoa from males carrying one t haplotype (t/+) exhibit mild motility deficits and a delay in penetration of the zona-free oocyte. The inversions of the t haplotypes contain several genes that cause or contribute to male sterility, at least some of which can be identified by analysis of mice carrying Mus spretus-Mus domesticus recombinant Chromosomes 17. The t haplotypes specify a number of sperm biochemical abnormalities, but these have not been related directly to defects in fertilization. In t/+ males, spermatozoa not bearing the t haplotype are defective in fertilization compared with t-bearing spermatozoa. The mechanism causing this is likely to involve haploid gene expression confined to the t-bearing spermatids. Since many genes situated in the region of the t haplotypes have human homologues, an understanding of t haplotype sterility in mice is expected to contribute significantly to our knowledge of the genetic basis for human sperm dysfunction. PMID- 9414480 TI - Regulatory mechanisms in acrosomal exocytosis. AB - Acrosomal exocytosis occurs after the binding of the spermatozoon to the zona pellucida of the oocyte via specific receptors. We suggest that the zona pellucida binds to at least two different receptors in the plasma membrane. One (R) is a Gi-coupled receptor that activates phospholipase C beta 1. The other (TK) is a tyrosine kinase receptor coupled to phospholipase C gamma. Binding to R would regulate adenylyl cyclase leading to an increase in cyclic adenosine monophosphate and protein kinase A activation. The protein kinase A activates a voltage-dependent Ca2+ channel in the outer acrosomal membrane that releases Ca2+ from the interior of the acrosome to the cytosol. This is the first (I), relatively small, rise in intracellular Ca2+ which leads to activation of the phospholipase C gamma. The products of phosphatidyl-inositol bisphosphate hydrolysis by phospholipase C, diacylglycerol and inositol-trisphosphate lead to protein kinase C translocation to the plasma membrane and its activation. Protein kinase C opens a voltage-dependent Ca2+ channel (L) in the plasma membrane, leading to the second (II), higher, increase in intracellular Ca2+ leading to acrosomal exocytosis. Spermine, a physiological constituent of the seminal plasma regulates sperm acrosomal exocytosis by modulating intracellular Ca2+ binding sites and phospholipase C activity. Spermine is rapidly incorporated into the sperm cells during ejaculation and temporarily inhibits premature capacitation and acrosome reaction. During the passage of the spermatozoon through the female genital tract, there is a progressive depletion of spermine from spermatozoa, so that capacitation and consequently the acrosomal exocytosis take place at the appropriate time, when the spermatozoon reaches the vicinity of the egg. PMID- 9414481 TI - Reproductive biology of seals. AB - The reproductive biology of seals is fascinating in many aspects. As in most mammals, the time of onset of puberty in seals is variable. Once sexually mature, most but not all seals are seasonally mono-oestrous, with highly synchronized breeding seasons. They have evolved as either terrestrial or aquatic copulators, although a few species mate in a variety of habitats. Their mating strategies are diverse, ranging from serial monogamy to extreme polygyny. Gestation in seals is characterized by an embryonic diapause, which is obligate in most species. Reactivation of the blastocyst is followed by a placental gestation. All species of seal require a terrestrial (including ice floes) habitat for parturition. Lactation differs between the two seal families: phocid seals have an intense period of maternal investment, during which the mothers fast; otariid seals have a prolonged lactation during which intense bouts of suckling are interspersed by days of separation from their pups while the mother forages at sea. Although the anatomy and functional morphology of seals has been well described, less is known of the endocrinology of reproduction. This is due mainly to the logistical difficulties that researchers experience in collecting serial samples from a species that is relatively difficult to handle. This article reviews the basic anatomy and physiology, and our current understanding of the comparative aspects of reproduction in seals. Reproductive behaviours as well as the influences of environmental factors, such as photoperiod, nutrition and xenobiotics, are also discussed. PMID- 9414482 TI - Regulatory DNA elements of phenobarbital-responsive cytochrome P450 CYP2B genes. AB - This article reviews recent progress in characterizing cis-acting DNA elements of the phenobarbital-inducible CYP2B genes. Whereas proximal DNA elements such as the C/EBP binding site regulate basal transcription activity, phenobarbital responsive enhancer activity is governed by the distal element (designated phenobarbital-responsive enhancer module, PBREM) residing about -2.3 kbp upstream from the transcription start site. Proximal elements are not required to enhance the phenobarbital-inducible transcription, since the PBREM can confer the inducibility to several heterologous promoters. Repression of the basal transcription by a negative element upstream of the -0.8 kbp region, however, may be necessary for the proper regulation of the CYP2B genes. PMID- 9414483 TI - Identification of the Yc1 glutathione S-transferase mRNA as the overexpressed species in a nitrogen mustard-resistant rat mammary carcinoma cell line. AB - Glutathione transferase (GSTs) have been shown to be overexpressed in a number of tumor cell lines selected for resistance to chemotherapeutic drugs and have been implicated in some studies of clinical specimens. In tumor cell lines selected for resistance to chemicals that alkylate DNA, the isoform most frequently overexpressed is GST-Yc, a member of the alpha class GSTs. To date, two variations of the cDNA designated Yc1 with subtle differences have been described, and Yc2 is shown to be clearly distinct. Transfection of a Yc1 cDNA constitutively expressed in rat liver into rat mammary cancer cells confers resistance to alkylators, however, to a lesser extent than is observed in the cells selected for resistance. It has therefore been widely suggested that the GST that is overexpressed in selected resistant cells represents a distinct and novel isoform. We have previously described a rat mammary carcinoma cell line (MLNr) that is resistant to alkylating agents, and overexpresses a GST with characteristics similar to GST-Yc1 and not Yc2. It has many features common to the several other GST-Yc overexpressing alkylator resistant cell lines. We have cloned the specific Yc cDNA overexpressed in MLNr and analyzed it in detail and found that it is identical to one of the previously reported Yc1 cDNAs, suggesting that there is no additional Yc gene specifically induced by nitrogen mustards. Another hypothesis to explain the difference in the level of resistance in selected versus GST-Yc transfected cells is the lack of concurrent increased glutathione (GSH) in the transfectants, which is a common feature in the selected resistant cells. Experiments in which we modulated GSH levels suggest that this is not likely. These studies add to our speculation that other mechanisms may be involved in alkylator resistance. PMID- 9414484 TI - Mechanism of inhibition of rat liver bilirubin UDP-glucuronosyltransferase by triphenylalkyl derivatives. AB - A series of potent and competitive inhibitors of UDP-glucuronosyltransferase derived from 7,7,7-triphenylheptanoic acid has been synthesized in order to probe the active site of the isozyme involved in the glucuronidation of the endogenous toxic compound, bilirubin IX alpha. Like triphenylalkylcarboxylic acids, triphenyl alcohols were found to be very effective competitive inhibitors of the reaction (Ki 12 to 180 microM). Superimposition of the best inhibitors with bilirubin by computer modeling showed a marked spatial similarity, which accounts for the observed competitive-type inhibition. The bulky triphenylmethyl moiety of the inhibitor superimposed well on the part of the bilirubin molecule containing three of the four pyrrole rings. In agreement, substitution of the triphenylmethyl moiety by planar structures such as fluorenyl or indenyl rings completely suppressed the inhibition. In addition, the weak inhibition exerted by the shortest carboxylic acids could be related to the higher acidity of these molecules. The inhibition potency depended on the acidity of the molecules; the more acidic, the less inhibitory, suggesting that the presence of a negative charge on the inhibitor molecule prevents bilirubin glucuronidation. Based on these results, a reaction mechanism for bilirubin glucuronidation is postulated. PMID- 9414485 TI - 2,3,7,8-Tetrachlorodibenzo-p-dioxin mechanism of action to reduce lipoprotein lipase activity in the 3T3-L1 preadipocyte cell line. AB - Lipoprotein lipase (LPL) is important in the process of triglyceride storage in adipose tissue. Depression of LPL activity in adipose tissue is associated with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-induced wasting syndrome and may have a role in the associated serum hyperlipidemia produced by TCDD. The 3T3-L1 cell line was used as an in vitro model, independent of hormonal, nutritional, or other interfering factors associated with in vivo studies, in order to systematically examine the mechanism of action of TCDD. TCDD produced a statistically significant (P < 0.05) time- and dose-dependent decrease in LPL activity. Results of experiments with Ah-receptor blockers and structure activity studies with different polychlorinated biphenyl (PCB) and dioxin congeners were consistent with reduction of LPL activity being mediated by the Ah receptor. Culturing of 3T3-L1 cells without glucose or with cytochalasin B, a blocker of facilitative glucose transporters (GLUT), was effective in reducing LPL activity (P < 0.05). TCDD did not further reduce LPL activity in cytochalasin B pretreated 3T3-L1 cells or in 3T3-L1 cells cultured in glucose-free media. Dexamethasone pretreatment, which is known to increase GLUT expression in 3T3-L1 cells, prevented the reduction of LPL activity by TCDD. Protein tyrosine kinase activities, assayed using gamma-32P-ATP and RR-SRC, a src specific peptide substrate, were significantly increased (P < 0.05) over control levels by both TCDD and glucose deprivation. Furthermore, results of experiments treating 3T3-L1 cells with either insulin, EGF, 8-Br-cAMP, TPA, or genistein, alone or in combination with TCDD, were generally consistent with the hypothesis that lowered intracellular glucose and altered cellular kinase activities may be involved in reduction of LPL activities by TCDD. Further work is needed to confirm and better understand the role protein phosphorylation plays in TCDD-mediated alteration of glucose disposition and LPL activity. In summary, TCDD reduced LPL activity in 3T3-L1 cells as seen in vivo. Manipulation of glucose transport through a number of experimental approaches produced changes in 3T3-L1 LPL activity consistent with results of previous investigators showing glucose to be a positive regulator of LPL activity and consistent with our hypothesis that TCDD-mediated reduction of glucose transport is an important factor in the down regulation of LPL activity by TCDD. PMID- 9414486 TI - The biochemical toxicology of 1,3-difluoro-2-propanol, the major ingredient of the pesticide gliftor: the potential of 4-methylpyrazole as an antidote. AB - Administration to rats of 1,3-difluoro-2-propanol (100 mg kg-1 body weight), the major ingredient of the pesticide gliftor, resulted in accumulation of citrate in the kidney after a 3 hour lag phase. 1,3-Difluoro-2-propanol was found to be metabolized to 1,3-difluoroacetone and ultimately to the aconitate hydratase inhibitor (-) erythrofluorocitrate and free fluoride. The conversion of 1,3 difluoro-2-propanol to 1,3-difluoroacetone was found to be catalyzed by an NAD(+) dependent alcohol dehydrogenase, while the defluorination was attributed to microsomal monooxygenase activity induced by phenobarbitone and inhibited by piperonyl butoxide. 4-Methylpyrazole was found to inhibit both of these processes in vitro and when administered (90 mg kg-1 body weight) to rats, 2 hours prior to 1,3-difluoro-2-propanol, eliminated signs of poisoning, prevented (-) erythrofluorocitrate synthesis, and markedly decreased citrate and fluoride accumulation in vivo. 4-Methylpyrazole also appeared to diminish (-) erythrofluorocitrate synthesis from fluoroacetate in vivo, and this was attributed to its capacity to inhibit malate dehydrogenase activity. The antidotal potential of 4-methylpyrazole and the potential for 1,3-difluoro-2 propanol to replace fluoroacetate (compound 1080) as a vertebrate pesticide is discussed. PMID- 9414487 TI - Superoxide dismutase-mimicking activities of dinuclear Cu(II) complexes with ligands containing a tetrathioether-tetraamino moiety. AB - The superoxide scavenging activities of copper(II) complexes with the ligands, 6,6'-methylene- bis(5'-amino-3',4'-benzo-2'-thiapentyl)-1,11-diamino- 2,3:9,10 dibenzo-4,8-dithiaundecane (H4L), and 6,6'- bis(5'-amino-3'4'-benzo-2' thiapentyl)-1,11-diamino- 2,3:9,10-dibenzo-4,8-dithiaundecane (H4L"), were investigated by xanthine-xanthine oxidase (X/XO) assays using nitroblue tetrazolium (NBT) as indicator molecule, and the results were compared with respect to the particular type of anion (ClO4, Cl, NO3) on the apical site of the copper(II) complexes. All of the complexes inhibited the reduction of NBT by superoxide radicals, with the [Cu2(L')](ClO4)2 complex exhibiting the highest scavenging activity against superoxide radicals among the complexes examined. The catalytic efficiency of the complexes for dismutation of superoxide radicals depends on the particular anion liganded to Cu(II) ion in the complexes, and the order of potency was observed to be ClO4 > Cl > NO3 in phosphate buffer at pH 7.40. The Cu(II)-H4L' complexes had the lowest IC50 and catalytic rate constant values indicating that the distorted geometry of the Cu(II)-H4L' complexes influence their catalytic activities for dismutation of superoxide radicals more efficiently. The difference in the activities of the complexes toward superoxide radicals can also be attributed to the nature of the anions on the apical site of the copper(II) complexes and the superoxide dismutase-like activity. PMID- 9414488 TI - Human liver paraoxonase (PON1): subcellular distribution and characterization. AB - The subcellular localization and different biochemical properties of a human hepatic microsomal enzyme that hydrolyses paraoxon (paraoxonase, PON1) were studied and compared to the paraoxon hydrolase activity found in human plasma as well as in rat liver and plasma. Having evaluated the influence of the postmortem interval by a parallel experiment performed in rats, we conclude that the paraoxonase activity was preferentially localized in the microsomal fraction. The enzyme reaction was optimized according to temperature, pH, buffer, ionic strength, substrate concentration, and enzyme protein concentration. The characterization of human liver paraoxonase included the study of optimum pH, pH stability, heat inactivation assays, and kinetic parameters (K(m) and Vmax). In addition, the enzyme activity showed an absolute requirement for exogenous calcium. The activity was lost after incubation with EDTA and partially restored by the addition of calcium; however, other metals assayed were not able to activate the human liver enzyme as did calcium. Our results support the possible identity between human plasma and liver paraoxonases. In spite of the technical difficulties of this study and the possible interference of the postmortem changes in the results, this article represents the first systematic approach to the characterization of human liver paraoxonase. PMID- 9414489 TI - Studying noncovalent protein complexes by electrospray ionization mass spectrometry. AB - Electrospray ionization mass spectrometry has been used to study protein interactions driven by noncovalent forces. The gentleness of the electrospray ionization process allows intact protein complexes to be directly detected by mass spectrometry. Evidence from the growing body of literature suggests that the ESI-MS observations for these weakly bound systems reflect, to some extent, the nature of the interaction found in the condensed phase. Stoichiometry of the complex can be easily obtained from the resulting mass spectrum because the molecular weight of the complex is directly measured. For the study of protein interactions, ESI-MS is complementary to other biophysical methods, such as NMR and analytical ultracentrifugation. However, mass spectrometry offers advantages in speed and sensitivity. The experimental variables that play a role in the outcome of ESI-MS studies of noncovalently bound complexes are reviewed. Several applications of ESI-MS are discussed, including protein interactions with metal ions and nucleic acids and subunit protein structures (quaternary structure). PMID- 9414490 TI - Ion-molecule reactions as probes of gas-phase structures of peptides and proteins. AB - A review with over 100 references describes the recent applications of ion molecule reactions to the study of gas-phase protonated peptides and proteins. The topic is focused specifically on the proton transfer and hydrogen-deuterium exchange reactions of amino acids, peptides, and proteins. A brief background is given of the various methods used for assigning proton affinities and gas-phase basicities. The methods used for measuring the kinetics of deuterium incorporation of charged ion in the presence of a background pressure of deuterating reagents are also described. Ion-molecule reactions are used to determine, among other things, the gas-phase basicities and proton affinities of amino acids, peptides, and proteins, the sites of protonation, intra- and intermolecular interactions, and conformational differences and changes in gas phase ionic species. Singly charged and multiply charged ions are both covered. PMID- 9414492 TI - Protein identification from 2-DE gels by MALDI mass spectrometry. PMID- 9414493 TI - [Nursing and professional development (I). Professional sociology--a theoretic assessment (I)]. PMID- 9414491 TI - Chiral recognition detected by fast atom bombardment mass spectrometry. AB - Detection of chiral recognition in various intermolecular interaction systems using mass spectrometry has become important for the modern fields of analytical chemistry, organic chemistry, and biochemistry due to the characteristic nature of the rapid method and the trace amount needed. This review presents the various methods for detecting and evaluating chiral recognition used primarily in fast atom bombardment mass spectrometry. Emphasis is put on fundamentals and applications of these methods for variously existing enantioselective intermolecular interaction systems. PMID- 9414494 TI - [Quality improvement in the care of patients with newly diagnosed diabetes mellitus]. PMID- 9414495 TI - [Pages from a primary nurse's diary]. PMID- 9414496 TI - [Model demonstration-unit instruction]. PMID- 9414497 TI - [The active consumer--consumer influence in the health system--which conditions?]. PMID- 9414499 TI - [Resume, analysis and critique of Evaluation of Nursing Education (I)]. PMID- 9414498 TI - [Care of the diabetic patient]. PMID- 9414500 TI - [Hospital admission interview in nursing anno 1996]. PMID- 9414501 TI - [Calling and profession]. PMID- 9414503 TI - [A health visitor's diary]. PMID- 9414502 TI - [Nursing and professional development (II). Nursing's development set against professional-theoretical framework]. PMID- 9414504 TI - [Spiritual care belongs with holistic nursing]. PMID- 9414505 TI - [Deeply declining years in its course]. PMID- 9414506 TI - [Resume, analysis and critique of Evaluation of Nursing Education (II)]. PMID- 9414507 TI - [9th Danish nursing symposium in Falkoner Center, Copenhagen]. PMID- 9414508 TI - [On suffering and suffering's justification]. PMID- 9414509 TI - [Language and profession]. PMID- 9414510 TI - [Nurses' education from a professional viewpoint]. PMID- 9414511 TI - [Education makes the difference--a study on the subject: does involvement in education increase concrete knowledge in diabetics?]. PMID- 9414512 TI - [Home care nursing is also geraniums and kerosene fumes]. PMID- 9414513 TI - [Patient progress in a stroke unit]. PMID- 9414514 TI - [Grundtvig's ideas in nursing. The Living, Soaring Word and its significance]. PMID- 9414515 TI - [There is no direct time for good ideas!--on reality's teaching conditions]. PMID- 9414516 TI - [The road ahead!--a project to help youth who attempted suicide]. PMID- 9414517 TI - [What is good nursing care and where does one learn to give good nursing care?]. PMID- 9414518 TI - [Historical development of the establishment of the Sister Marie Dalgaard's Foundation for nursing research and development]. PMID- 9414519 TI - [Resume, analysis and critique of Evaluation of Nurse's Education (I)]. PMID- 9414520 TI - [Profession and science]. PMID- 9414521 TI - [Health visitors activities as seen from a professional viewpoint]. PMID- 9414522 TI - [Persistence--the greatest challenge in a quality assurance project]. PMID- 9414523 TI - [Joys and regrets in development activities]. PMID- 9414524 TI - [Jest or fraud--science between formalization and social construction]. PMID- 9414525 TI - [Suffering in women behind the mask]. PMID- 9414526 TI - [Youth in nursing--practical reality in health and nursing care, where are the role models?]. PMID- 9414527 TI - [Confidential information doctrine in practice--everyday situation from clinical practice]. PMID- 9414528 TI - [Inadequate planning is expensive!]. PMID- 9414529 TI - [Profession and leadership (educ)]. PMID- 9414530 TI - [Professional dilemmas in nursing]. PMID- 9414532 TI - [To be present--a professional nurse's expression]. PMID- 9414531 TI - [Hospital system in society--with special reference to the development of team functions as a mode of work]. PMID- 9414533 TI - [When songs have wings]. PMID- 9414534 TI - [Psychotherapy in suicidal youth]. PMID- 9414535 TI - [Youth in nursing--building bridges between theory and practice in future nursing]. PMID- 9414536 TI - [Clinical dropout in nurses' education]. PMID- 9414537 TI - [Personal nursing care]. PMID- 9414538 TI - [Consumer classification and nursing load in integrated home care arrangements]. PMID- 9414539 TI - [Men and methods in clinical health care activities--from nursing process to cross-professional team method]. PMID- 9414540 TI - Home parenteral nutrition: a systematic review. AB - OBJECTIVES: The objective of this Review was to locate, appraise and summarise evidence from scientific studies on home parenteral nutrition (HPN) in order to answer specific research questions on the effectiveness of this technology. The following questions were asked. What patients have received HPN? What has been the experience of patients on HPN programmes? How have HPN programmes been organised, and what techniques and equipment have been used, and to what effect? What comparative information is available on effectiveness? What evidence exists for the cost-effectiveness of HPN? What questions about the provision of HPN could be answered with additional research, and what studies would be most suitable? DATA SOURCES: A comprehensive list of studies was provided by an extensive search of electronic databases (including MEDLINE, Embase, Science Citation Index, Uncover, Cinahl, Caredata, Food Science and Technology Abstracts, NTIS, Pascal, Psychlit, and Economic Literature Index), relevant journals (including Journal of Parenteral and Enteral Nutrition, Clinical Nutrition, American Journal of Clinical Nutrition, Nutrition, Clinical Gastroenterology, Nutrition Reviews, Annals of Nutrition and Metabolism, Nutrition and Cancer, Nutrition and Health, and Journal of Paediatric Nutrition and Metabolism), and scanning of reference lists, as well as other search strategies outlined in the protocol. STUDY SELECTION: Studies relevant to the questions were selected. The inclusion criteria were fairly broad because of the quality of the studies located. DATA EXTRACTION: Data extraction forms were used to collect data from studies included in the review. The data was checked by a second researcher to reduce error. DATA SYNTHESIS: Quantitative analysis was difficult owing to the type of studies located. The data is discussed in a qualitative manner. Where complication rates have been given, we have attempted to combine the results in a quantitative manner. RESULTS: The age and sex of patients on HPN varies according to the underlying disease but, on the whole, patients are young (see Tables 4a and 4b). There are trends showing an increased use of the technology at the extremes of the age range. There are marked differences between countries on the underlying diseases for which HPN is indicated. For example, many more patients with an underlying malignancy are treated in Italy and the USA than in the UK (40 67% versus 8%). Morbidity rates for the majority of patients are acceptable (see Table 8), the complications tend to be related to the central venous catheter. It is fairly clear that a minority of patients are susceptible to recurrent problems and that many patients have very few complications. The mortality rate for HPN patients (see Table 10) was good for those patients with benign underlying disease (for example, 5% of Crohn's HPN patients die per year), and there are very few reports of patients dying from complications of the technology. The survival of those with malignant disease and AIDS is poor, almost all having died from the underlying disease at one year; despite this, most programme growth worldwide is due to an increase in the numbers of patients with these diagnoses (see Table 5). Quality of life is reasonable for patients with benign disease (see Table 9); no studies were found that examined the quality of life of HPN patients with malignant disease. Economic analysis shows that the cost of HPN treatment is cheaper than the alternative of in-patient care (see Table 18). There is a paucity of comparative studies examining different aspects of the technology, and this accounted for the majority of gaps in the evidence. CONCLUSIONS: The use of HPN for benign intestinal failure is supported by evidence from the scientific studies located. There are, however, large gaps in the evidence, particularly relating to the use of HPN in malignant disease and AIDS. A programme of research is suggested at the end of this review. PMID- 9414541 TI - Diagnosis, management and screening of early localised prostate cancer. AB - The incidence of prostate cancer is rising worldwide, caused mainly by demographic factors, particularly the increasingly elderly population and, more importantly, the increasing number of cases identified following prostate specific antigen (PSA) testing. It is commonly quoted that many more men die with prostate cancer than of it. Autopsy/post-mortem studies show that while a very high proportion of elderly men have histological evidence of the disease, a much smaller proportion develop clinically apparent cancer. The natural history of prostate cancer is poorly understood, but progression appears to be related to stage and grade of tumour. Prostate cancer can be diagnosed by digital rectal examination (DRE), serum PSA test, and/or transrectal ultrasound (TRUS), with confirmation by biopsy. Each test identifies a proportion of cancers, with higher rates of detection when they are used in combination. The tests are also used to determine which tumours are localised within the prostate and are, thus, potentially treatable. Unfortunately, clinical staging is unreliable, with approximately one half of all tumours upstaged following surgery. Three major treatment options are available for localised prostate cancer: radical prostatectomy, radical radiotherapy and conservative management (involving monitoring and treatment of symptoms). Although radical treatment rates are rising, good quality evidence concerning their comparative effectiveness and cost effectiveness is lacking. Observational studies of highly selected patient groups suggests that there may be a slightly lower mortality rate following radical treatments compared with conservative management, but there has been very little research into treatment complications and quality of life of men after any of the treatments. In the past, investigations of prostate cancer were reserved largely for patients exhibiting symptoms, but the introduction of the PSA test has opened up the possibility of screening healthy men for the disease. Observational studies suggest that DRE and PSA, combined with TRUS and biopsy, can identify localised prostate cancer in 3-5% of men, although the tests do result in a number of false positives and negatives. Major questions remain concerning the natural history of the disease, potential costs (financial, social and psychological) of a screening programme, and the effectiveness and cost effectiveness of treatments for localised disease. The lack of good quality data and the strength of these concerns means that population screening for prostate cancer cannot be recommended. PMID- 9414542 TI - The diagnosis, management, treatment and costs of prostate cancer in England and Wales. PMID- 9414543 TI - Screening for fragile X syndrome. AB - BACKGROUND AND AIM OF REVIEW. In 1991, the gene responsible for fragile X syndrome, a common cause of learning disability, was discovered. As a result, diagnosis of the disorder has improved and its molecular genetics are now understood. This report seems to provide the information needed to decide whether to use DNA testing to screen for the disorder. HOW THE RESEARCH WAS CONDUCTED. A literature search of electronic reference databases of published and 'grey' literature was undertaken together with hand searching of the most recent publications. RESEARCH FINDINGS. NATURAL HISTORY. Physical characteristics of fragile X syndrome include facial atypia, joint laxity and, in boys, macro orchidism. Most affected males have moderate-to-severe learning disabilities with IQs under 50 whereas most females have borderline IQs of 70-85. Behavioural problems are similar to those seen with autism and attention-deficit disorders. Although fragile X syndrome is not curable there are a number of medical, educational, psychological and social interventions that can improve the symptoms. About 6% of those with learning disabilities tested in institutions have fragile X syndrome. Population prevalence figures are 1 in 4000 in males and 1 in 8000 in females. GENETICS. The disorder is caused by a mutation in a gene on the X chromosome which includes a trinucleotide repeat sequence. The mutation is characterized by hyper-expansion of the repeat sequence leading to down regulation of the gene. In males an allele with repeat size in excess of 200, termed a full mutation (FM), is always associated with the affected phenotype, whereas in females only half are affected. Individuals with alleles having repeat size in the range 55-199 are unaffected but in females the sequence is heritably unstable so that it is at high risk of expansion to an FM in her offspring. This allele is known as a pre-mutation (PM) to contrast it with the FM found in the affected individual. No spontaneous expansions directly from a normal allele to an FM have been observed. SCREENING STRATEGIES. The principal aims of screenng for fragile X syndrome is to reduce the birth prevalence of the disorder, by prenatal diagnosis and selective termination of pregnancy, or by reducing the number of pregnancies in women who have the FM or PM alleles. Possible screening strategies are: routine antenatal testing of apparently low risk pregnancies, preconceptual testing of young women, and systematic testing in affected families ('cascade' screening). A secondary aim is to bring forward the diagnosis of affected individuals so that they might benefit from early treatment. Active paediatric screening and neonatal screening could achieve this but there is no direct evidence of any great benefit from early diagnosis. SCREENING TESTS. Cytogenetic methods are unsuitable for screening purposes. Southern blotting of genomic DNA can be used but is inaccurate in measuring the size of small PMs, there is a long laboratory turnaround time, and it is relatively expensive. The best protocol is to amplify the DNA using polymerase chain reaction on all samples, and when there is a possible failure to amplify, a Southern blot.(ABSTRACT TRUNCATED) PMID- 9414544 TI - Translational regulation in the chloroplast. PMID- 9414545 TI - Characterization of the common bean uricase II and its expression in organs other than nodules. AB - Uricase II is a purine metabolic enzyme highly induced in root nodules during the symbiosis established between legumes and bacteria of the genera Rhizobium and Bradyrhizobium. Here we describe the characterization of bean (Phaseolus vulgaris) nodule uricase II cDNA and show that uricase II is encoded by a single gene in the bean genome. This gene is also expressed in cotyledons, roots, and hypocotyls during bean seedling establishment, and an anti-uricase antibody recognizes the protein in different seedling organs. Uricase II has also been found in Leucaena leucocephala seedlings, suggesting that it participates during seedling establishment in legumes that do not transport ureides. A 50-kD polypeptide that is detected by the anti-uricase antibody is found in cotyledons during seedling development. This higher-molecular-mass form is also detected in developing roots and hypocotyls but not in nodules. In situ hybridization experiments in root seedlings showed uricase II transcripts in the metaxylem parenchyma cells and phloem fibers of the vascular system. PMID- 9414546 TI - Cellular localization of Arabidopsis xyloglucan endotransglycosylase-related proteins during development and after wind stimulation. AB - A gene family encoding xyloglucan endotransglycosylase (XET)-related proteins exists in Arabidopsis. TCH4, a member of this family, is strongly up-regulated by environmental stimuli and encodes an XET capable of modifying cell wall xyloglucans. To investigate XET localization we generated antibodies against the TCH4 carboxyl terminus. The antibodies recognized TCH4 and possibly other XET related proteins. These data indicate that XETs accumulate in expanding cell, at the sites of intercellular airspace formation, and at the bases of leaves, cotyledons, and hypocotyls. XETs also accumulated in vascular tissue, where cell wall modifications lead to the formation of tracheary elements and sieve tubes. Thus, XETs may function in modifying cell walls to allow growth, airspace formation, the development of vasculature, and reinforcement of regions under mechanical strain. Following wind stimulation, overall XET levels appeared to decrease in the leaves of wind-stimulated plants. However, consistent with an increase in TCH4 mRNA levels following wind, there were regions that showed increased immunoreaction, including sites around cells of the pith parenchyma, between the vascular elements, and within the epidermis. These results indicate that TCH4 may contribute to the adaptive changes in morphogenesis that occur in Arabidopsis following exposure to mechanical stimuli. PMID- 9414547 TI - The aberrant cell walls of boron-deficient bean root nodules have no covalently bound hydroxyproline-/proline-rich proteins. AB - B-deficient bean (Phaseolus vulgaris L.) nodules examined by light microscopy showed dramatic anatomical changes, mainly in the parenchyma region. Western analysis of total nodule extracts examined by sodium dodecyl sulfate polyacrylamide gel electrophoresis showed that one 116-kD polypeptide was recognized by antibodies raised against hydroxyproline-rich glycoproteins (HRGPs) from the soybean (Glycine max) seed coat. A protein with a comparable molecular mass of 116 kD was purified from the cell walls of soybean root nodules. The amino acid composition of this protein is similar to the early nodulin (ENOD2) gene. Immunoprecipitation of the soybean ENOD2 in vitro translation product showed that the soybean seed coat anti-HRGP antibodies recognized this early nodulin. Furthermore, we used these antibodies to localize the ENOD2 homolog in bean nodules. Immunocytochemistry revealed that in B-deficient nodules ENOD2 was absent in the walls of the nodule parenchyma. The absence of ENOD2 in B-deficient nodules was corroborated by performing hydroxyproline assays. Northern analysis showed that ENOD2 mRNA is present in B-deficient nodules; therefore, the accumulation of ENOD2 is not affected by B deficiency, but its assembly into the cell wall is. B-deficient nodules fix much less N2 than control nodules, probably because the nodule parenchyma is no longer an effective O2 barrier. PMID- 9414548 TI - Association of caffeoyl coenzyme A 3-O-methyltransferase expression with lignifying tissues in several dicot plants. AB - Caffeoyl coenzyme A 3-O-methyltransferase (CCoAOMT) was previously shown to be associated with lignification in both in vitro tracheary elements (TEs) and organs of zinnia (Zinnia elegans). However, it is not known whether this is a general pattern in dicot plants. To address this question, polyclonal antibodies against zinnia recombinant CCoAOMT fusion protein were raiseed and used for immunolocalization in several dicot plants. The antibodies predominantly recognized a protein band with a molecular mass of 28 kD on western analysis of tissue extracts from zinnia, forsythia (Forsythia suspensa), tobacco (Nicotiana tabacum), alfalfa (Medicago sativa), and soybean (Glycine max). Western analyses showed that the accumulation of CCoAOMT protein was closely correlated with lignification in in vitro TEs of zinnia. Immunolocalization results showed that CCoAOMT was localized in developing TEs of young zinnia stems and in TEs, xylem fibers, and phloem fibers of old stems. CCoAOMT was also found to be specifically associated with all lignifying tissues, including TEs, xylem fibers, and phloem fibers in stems of forsythia, tobacco, alfalfa, soybean, and tomato (Lycopersicon esculentum). The presence of CCoAOMT was evident in xylem ray parenchyma cells of forsythia, tobacco, and tomato. In forsythia and alfalfa, pith parenchyma cells next to the vascular cylinder were lignified. Accordingly, marked accumulation of CCoAOMT in these cells was observed. Taken together, these results showed a close association of CCoAOMT expression with lignification in dicot plants. This supports the hypothesis that the CCoAOMT-mediated methylation branch is a general one in lignin biosynthesis during normal growth and development in dicot plants. PMID- 9414549 TI - Evidence that heat and ultraviolet radiation activate a common stress-response program in plants that is altered in the uvh6 mutant of Arabidopsis thaliana. AB - The uvh6 mutant of Arabidopsis was previously isolated in a screen for increased sensitivity to ultraviolet (UV) radiation. uvh6 mutant plants were killed by incubation at 37 degrees C for 4 d, a treatment not lethal to wild-type plants. Furthermore, under permissive conditions, uvh6 plants were yellow-green with an approximately one-third lower chlorophyll content. Genetic analysis of the uvh6 mutant strongly suggested that all three mutant phenotypes were due to mutation at the same genetic locus. To understand UVH6 function more fully, the response of wild-type plants to growth at elevated temperatures and exposure to UV radiation was analyzed. Wild-type plants grown at 30 degrees C were as UV hypersensitive and yellow-green as uvh6 mutant plants grown at 24 degrees C. Mutant uvh6 plants induced heat-shock protein HSP21 at a lower threshold temperature than wild-type plants, indicating that the uvh6 mutant was exhibiting signs of heat stress at a 4 to 5 degrees C lower temperature than wild-type plants. We propose the UV damage and heat induce a common stress response in plants that leads to tissue death and reduced chloroplast function, and that the UVH6 product is a negative regulator of this response. PMID- 9414550 TI - Sequencing, genomic organization, and preliminary promoter analysis of a black cherry (R)-(+)-mandelonitrile lyase gene. AB - The flavoprotein (R)-(+)-mandelonitrile lyase (MDL; EC 4.1.2.10) plays a key role in cyanogenesis in rosaceous stone fruits. An MDL gene (mdl3) and its corresponding cDNA (MDL3) were isolated from black cherry (Prunus serotina) and characterized. The mdl3 gene contains 2292 bp of the 5' flanking region, the entire coding region, and 300 bp of the 3' flanking region. The coding region is interrupted by three short introns, of which one possesses the usual GC-AG splice junction dinucleotides. This gene encodes a polypeptide of 573 amino acids that includes a putative signal sequence, 13 potential N-glycosylation sites, and a presumptive flavin adenine dinucleotide-binding site. To determine whether the 5' flanking region of the mdl3 gene is capable of driving MDL expression, it was fused to the beta-glucuronidase reporter gene for Agrobacterium-mediated transformation into tobacco. Matching endogenous MDL expression patterns, beta glucuronidase staining was observed in maturing embryos and seeds; it also occurred in postembryonic tissues, especially in association with vascular tissues. After developing a homologous transient transformation system to facilitate identification of putative regulatory sequences, we demonstrated that 125 bp (-107 to +18) of the 5' flanking sequence of the mdl3 gene is sufficient for MDL expression in protoplasts derived from immature black cherry embryos. PMID- 9414551 TI - Biochemical and molecular biological characterization of CAC2, the Arabidopsis thaliana gene coding for the biotin carboxylase subunit of the plastidic acetyl coenzyme A carboxylase. AB - The biotin carboxylase subunit of the heteromeric chloroplastic acetyl-coenzyme A carboxylase (ACCase) of Arabidopsis thaliana is coded by a single gene (CAC2), which is interrupted by 15 introns. The cDNA encodes a deduced protein of 537 amino acids with an apparent N-terminal chloroplast-targeting transit peptide. Antibodies generated to a glutathione S-transferase-CAC2 fusion protein react solely with a 51-kD polypeptide of Arabidopsis; these antibodies also inhibit ACCase activity in extracts of Arabidopsis. The entire CAC2 cDNA sequence was expressed in Escherichia coli and the resulting recombinant biotin carboxylase was enzymatically active in carboxylating free biotin. The catalytic properties of the recombinant biotin carboxylase indicate that the activity of the heteromeric ACCase may be regulated by light-/dark-induced changes in stromal pH. The CAC2 gene is maximally expressed in organs and tissues that are actively synthesizing fatty acids for membrane lipids or oil deposition. The observed expression pattern of CAC2 mirrors that previously reported for the CAC1 gene (J. K. Choi, F. Yu, E.S. Wurtele, B.J. Nikolau [1995] Plant Physiol 109: 619-625; J. Ke, J.-K. Choi, M. Smith, H.T. Horner, B.J. Nikolau, E.S. Wurtele [1997] Plant Physiol 113: 357-365), which codes for the biotin carboxyl carrier subunit of the heteromeric ACCase. This coordination is probably partially established by coordinate transcription of the two genes. This hypothesis is consistent with the finding that the CAC2 and CAC1 gene promoters share a common set of sequence motifs that may be important in guiding the transcription of these genes. PMID- 9414552 TI - Identification of proliferation-induced genes in Arabidopsis thaliana. Characterization of a new member of the highly evolutionarily conserved histone H2A.F/Z variant subfamily. AB - The changes in gene expression associated with the reinitiation of cell division and subsequent progression through the cell cycle in Arabidopsis thaliana cell suspension cultures were investigated. Partial synchronization of cells was achieved by a technique combining phosphate starvation and a transient treatment with the DNA replication inhibitor aphidicolin. Six cDNAs corresponding to genes highly induced in proliferating cells and showing cell-cycle-regulated expression were obtained by the mRNA differential display technique. Full-length cDNA clones (cH2BAt and cH2AvAt) corresponding to two of the display products were subsequently isolated. The cH2BAt clone codes for a novel histone H2B protein, whereas the cH2AvAt cDNA corresponds to a gene encoding a new member of the highly conserved histone H2A.F/Z subfamily of chromosomal proteins. Further studies indicated that H2AvAt mRNA expression is tightly correlated with cell proliferation in cell-suspension cultures, and that closely related analogs of the encoded protein exist in Arabidopsis. The implications of the conservation of histone H2A.F/Z variants in plants are discussed. PMID- 9414553 TI - Subcellular localization of celery mannitol dehydrogenase. A cytosolic metabolic enzyme in nuclei. AB - Mannitol dehydrogenase (MTD) is the first enzyme in mannitol catabolism in celery (Apium graveolens L. var dulce [Mill] Pers. cv Florida 638). Mannitol is an important photoassimilate, as well as providing plants with resistance to salt and osmotic stress. Previous work has shown that expression of the celery Mtd gene is regulated by many factors, such as hexose sugars, salt and osmotic stress, and salicylic acid. Furthermore, MTD is present in cells of sink organs, phloem cells, and mannitol-grown suspension cultures. Immunogold localization and biochemical analyses presented here demonstrate that celery MTD is localized in the cytosol and nuclei. Although the cellular density of MTD varies among different cell types, densities of nuclear and cytosolic MTD in a given cell are approximately equal. Biochemical analyses of nuclear extracts from mannitol-grown cultured cells confirmed that the nuclear-localized MTD is enzymatically active. The function(s) of nuclear-localized MTD is unknown. PMID- 9414554 TI - Identification of active-site histidine residues of a self-incompatibility ribonuclease from a wild tomato. AB - The style component of the self-incompatibility (S) locus of the wild tomato Lycopersicon peruvianum (L.) Mill. is an allelic series of glycoproteins with ribonuclease activity (S-RNases). Treatment of the S3-RNase from L. peruvianum with iodoacetate at pH 6.1 led to a loss of RNase activity. In the presence of a competitive inhibitor, guanosine 3'-monophosphate (3'-GMP), the rate of RNase inactivation by iodoacetate was reduced significantly. Analysis of the tryptic digestion products of the iodoacetate-modified S-RNase by reversed-phase high performance liquid chromatography and electrospray-ionization mass spectrometry showed that histidine-32 was preferentially modified in the absence of 3'-GMP. Histidine-88 was also modified, but this occurred both in the presence and absence of 3'-GMP, suggesting that this residue is accessible when 3'-GMP is in the active site. Cysteine-150 was modified by iodoacetate in the absence of 3' GMP and, to a lesser extent, in its presence. The results are discussed with respect to the related fungal RNase T2 family and the mechanism of S-RNase action. PMID- 9414555 TI - Immunolocalization of PsNLEC-1, a lectin-like glycoprotein expressed in developing pea nodules. AB - The pea (Pisum sativum) nodule lectin gene PsNlec1 is a member of the legume lectin gene family that is strongly expressed in infected pea nodule tissue. A full-length cDNA sequence of PsNlec1 was expressed in Escherichia coli and a specific antiserum was generated from the purified protein. Immunoblotting of material from isolated symbiosomes revealed that the glycoprotein was present in two antigenic isoforms, PsNLEC-1A and PsNLEC-1B. The N-terminal sequence of isoform A showed homology to an eight-amino acid propeptide sequence previously identified from the cDNA sequence of isoform B. In nodule homogenates the antiserum recognized an additional fast-migrating band, PsNLEC-1C. Fractionation studies indicated that PsNLEC-1C was associated with a 100,000 g nodule membrane fraction, suggesting an association with cytoplasmic membrane or vesicles. Immunogold localization in pea nodule tissue sections demonstrated that the PsNLEC-1 antigen was present in the symbiosome compartment and also in the vacuole but revealed differences in distribution between infected host cells in different parts of the nodule. These data suggest that PsNLEC-1 is subject to posttranslational modification and that the various antigenic isoforms can be used to monitor membrane and vesicle targeting during symbiosome development. PMID- 9414556 TI - Signal transduction in the carnivorous plant Sarracenia purpurea. Regulation of secretory hydrolase expression during development and in response to resources. AB - Carnivory in plants has developed as an evolutionary adaptation to nutrient-poor environments. A significant investment of the resources of a carnivorous plant is committed to producing the traps, attractants, and digestive enzymes needed for the carnivory. The cost:benefit ratio of carnivory can be improved by either maximizing the prey capture rate or by reducing the metabolic commitment toward carnivory. Using the pitcher plant Sarracenia purpurea, we have investigated whether the expression of the hydrolytic enzymes needed for digestion is regulated in response to the presence of prey. Expression of protease, RNase, nuclease, and phosphatase activities could be induced in the fluid of nonactive traps by the addition of nucleic acids, protein, or reduced nitrogen, suggesting that hydrolase expression is induced upon perception of the appropriate chemical signal. Hydrolase expression was also developmentally controlled since expression commenced upon opening of a trap, increased for several days, and in the absence of prey largely ceased within 2 weeks. Nevertheless, the traps remained competent to induce expression in response to the appropriate signals. These data suggest that in young traps hydrolase expression is developmentally regulated, which is later replaced by a signal transduction mechanism, and they demonstrate the ability of a carnivorous species to respond to the availability of resources. PMID- 9414557 TI - Thermal protection of the oxygen-evolving machinery by PsbU, an extrinsic protein of photosystem II, in Synechococcus species PCC 7002. AB - The evolution of oxygen is the reaction that is the most susceptible to heat in photosynthesis. We showed previously that, in the cyanobacterium Synechococcus sp. PCC 7002, some protein factors located on the thylakoid membranes are involved in the stabilization of this reaction against heat-induced inactivation, and we identified cytochrome C550 as one such factor (Y. Nishiyama, H. Hayashi, T. Watanabe, N. Murata [1994] Plant Physiol 105: 1313-1319). In the present study we purified another protein that appears to be essential for the stabilization of the oxygen-evolving machinery. The purified protein had an apparent molecular mass of 13 kD, and the gene encoding the 13-kD protein was cloned from Synechococcus sp. PCC 7002 and sequenced. The deduced amino acid sequence revealed that the protein was homologous to PsbU, an extrinsic protein of the photosystem II complex, which has been found in thermophilic species of cyanobacteria. Western analysis showed that the level of PsbU in thylakoid membranes was constant, regardless of the growth temperature. Our studies indicate that PsbU, a constituent of the photosystem II complex, protects the oxygen-evolving machinery against heat-induced inactivation. PMID- 9414558 TI - Protein repair L-isoaspartyl methyltransferase in plants. Phylogenetic distribution and the accumulation of substrate proteins in aged barley seeds. AB - Protein L-isoaspartate (D-aspartate) O-methyltransferases (MTs; EC 2.1.1.77) can initiate the conversion of detrimental L-isoaspartyl residues in spontaneously damaged proteins to normal L-aspartyl residues. We detected this enzyme in 45 species from 23 families representing most of the divisions of the plant kingdom. MT activity is often localized in seeds, suggesting that it has a role in their maturation, quiescence, and germination. The relationship among MT activity, the accumulation of abnormal protein L-isoaspartyl residues, and seed viability was explored in barley (Hordeum vulgare cultivar Himalaya) seeds, which contain high levels of MT. Natural aging of barley seeds for 17 years resulted in a significant reduction in MT activity and in seed viability, coupled with increased levels of "unrepaired" L-isoaspartyl residues. In seeds heated to accelerate aging, we found no reduction of MT activity, but we did observe decreased seed viability and the accumulation of isoaspartyl residues. Among populations of accelerated aged seed, those possessing the highest levels of L isoaspartyl-containing proteins had the lowest germination percentages. These results suggest that the MT present in seeds cannot efficiently repair all spontaneously damaged proteins containing altered aspartyl residues, and their accumulation during aging may contribute to the loss of seed viability. PMID- 9414560 TI - Specific localization of the respiratory alternative oxidase in meristematic and xylematic tissues from developing soybean roots and hypocotyls. AB - We used tissue printing and specific immunostaining to examine the localization of the alternative oxidase (AOX) protein in correlation with measurements of AOX capacity. Selected root and hypocotyl regions were analyzed during the first 14 d of growth. It is shown that AOX protein is localized in the apical meristem and in developing xylem. The temporal pattern of expression is coincident with the evolution of AOX capacity. Data suggest that AOX expression is linked to xylem differentiation. Since heat is a major product of the alternative pathway, we speculate that thermogenesis is implicated in morphogenesis. PMID- 9414559 TI - Actin filaments of guard cells are reorganized in response to light and abscisic acid. AB - We recently showed that treatment with actin antagonists perturbed stomatal behavior in Commelina communis L. leaf epidermis and therefore suggested that dynamic changes in actin are necessary for signal responses in guard cells (M. Kim, P.K. Hepler, S.O. Eun, K.-S. Ha, Y. Lee [1995] Plant Physiol 109: 1077 1084). Here we show that actin filaments of guard cells, visualized by immunofluorescence microscopy, change their distribution in response to physiological stimuli. When stomata were open under white-light illumination, actin filaments were localized in the cortex of guard cells, arranged in a pattern that radiates from the stomatal pore. In marked contrast, for guard cells of stomata closed by darkness or by abscisic acid, the actin organization was characterized by short fragments randomly oriented and diffusely labeled along the pore site. Upon abscisic acid treatment, the radial pattern of actin arrays in the illuminated guard cells began to disintegrate within a few minutes and was completely disintegrated in the majority of labeled guard cells by 60 min. Unlike actin filaments, microtubules of guard cells retained an unaltered organization under all conditions tested. These results further support the involvement of actin filaments in signal transduction pathways of guard cells. PMID- 9414561 TI - Antisense inhibition of the photosystem I antenna protein Lhca4 in Arabidopsis thaliana. AB - The function of Lhca4, a gene encoding the photosystem 1 type IV chlorophyll a/b binding protein complex in Arabidopsis, was investigated using antisense technology. Lhca4 protein was reduced in a number of mutant lines and abolished in one. The inhibition of protein was not correlated with the inhibition of mRNA. No depletion of Lhca1 was observed, but the low-temperature fluorescence emission spectrum was drastically altered in the mutants. The emission maximum was blue shifted by 6 nm, showing that chlorophyll molecules bound to Lhca4 are responsible for most of the long-wavelength fluorescence emission. Some mutants also showed an unexplainable delay in flowering time and an increase in seed weight. PMID- 9414562 TI - Fluence and wavelength requirements for Arabidopsis CAB gene induction by different phytochromes. AB - The roles of different phytochromes have been investigated in the photoinduction of several chlorophyll a/b-binding protein genes (CAB) of Arabidopsis thaliana. Etiolated seedlings of the wild type, a phytochrome A (PhyA) null mutant (phyA), a phytochrome B (PhyB) null mutant (phyB), and phyA/phyB double mutant were exposed to monochromatic light to address the questions of the fluence and wavelength requirements for CAB induction by different phytochromes. In the wild type and the phyB mutant, PhyA photoirreversibly induced CAB expression upon irradiation with very-low-fluence light of 350 to 750 nm. In contrast, using the phyA mutant, PhyB photoreversibly induced CAB expression with low-fluence red light. The threshold fluences of red light for PhyA- and PhyB-specific induction were about 10 nmol m-2 and 10 mumol m-2, respectively. In addition, CAB expression was photoreversibly induced with low-fluence red light in the phyA/phyB double mutant, revealing that another phytochrome(s) (PhyX) regulated CAB expression in a manner similar to PhyB. These data suggest that plants utilize different phytochromes to perceive light of varying wave-lengths and fluence, and begin to explain how plants respond so exquisitely to changing light in their environment. PMID- 9414563 TI - RNase activity prevents the growth of a fungal pathogen in tobacco leaves and increases upon induction of systemic acquired resistance with elicitin. AB - The hypersensitive response and systemic acquired resistance (SAR) can be induced in tobacco (Nicotiana tabacum L.) plants by cryptogein, an elicitin secreted by Phytophthora cryptogea. Stem application of cryptogein leads to the establishment of acquired resistance to subsequent leaf infection with Phytophthora parasitica var nicotianae, the agent of the tobacco black shank disease. We have studied early events that occur after the infection and show here that a tobacco gene encoding the extracellular S-like RNase NE is expressed in response to inoculation with the pathogenic fungus. Upon induction of SAR with cryptogein, the accumulation of NE transcripts coincided with a rapid induction of RNase activity and with the increase in the activity of at least two different extracellular RNases. Moreover, exogenous application of RNase activity in the extracellular space of leaves led to a reduction of the fungus development by up to 90%, independently of any cryptogein treatment and in the absence of apparent necrosis. These results indicate that the up-regulation of apoplastic RNase activity after inoculation could contribute to the control of fungal invasion in plants induced to SAR with cryptogein. PMID- 9414564 TI - Ribulose-1,5-bisphosphate carboxylase/oxygenase activase deficiency delays senescence of ribulose-1,5-bisphosphate carboxylase/oxygenase but progressively impairs its catalysis during tobacco leaf development. AB - Transgenic tobacco (Nicotiana tabacum L. cv W38) plants with an antisense gene directed against the mRNA of ribulose-1,5-biphosphate carboxylase/oxygenase (Rubisco) activase grew more slowly than wild-type plants in a CO2-enriched atmosphere, but eventually attained the same height and number of leaves. Compared with the wild type, the anti-activase plants had reduced CO2 assimilation rates, normal contents of chlorophyll and soluble leaf protein, and much higher Rubisco contents, particularly in older leaves. Activase deficiency greatly delayed the usual developmental decline in Rubisco content seen in wild type leaves. This effect was much less obvious in another transgenic tobacco with an antisense gene directed against chloroplast-located glyceraldehyde-3-phosphate dehydrogenase, which also had reduced photosynthetic rates and delayed development. Although Rubisco carbamylation was reduced in the anti-activase plants, the reduction was not sufficient to explain the reduced photosynthetic rate of older anti-activase leaves. Instead, up to a 10-fold reduction in the catalytic turnover rate of carbamylated Rubisco in vivo appeared to be the main cause. Slower catalytic turnover by carbamylated Rubisco was particularly obvious in high-CO2-grown leaves but was also detectable in air-grown leaves. Rubisco activity measured immediately after rapid extraction of anti-activase leaves was not much less than that predicted from its degree of carbamylation, ruling out slow release of an inhibitor from carbamylated sites as a major cause of the phenomenon. Nor could substrate scarcity or product inhibition account for the impairment. We conclude that activase must have a role in vivo, direct or indirect, in promoting the activity of carbamylated Rubisco in addition to its role in promoting carbamylation. PMID- 9414565 TI - Identification and characterization of a novel arabinoxylanase from wheat flour. AB - An endogenous wheat (Triticum aestivum) flour endoxylanase was purified to homogeneity from a crude wheat flour extract by ammonium sulfate precipitation and cation-exchange chromatography. The 30-kD protein had an isoelectric point of 9.3 or higher. A sequence of 19 amino acids at the NH2 terminus showed 84.2% identity with an internal sequence of 15-kD grain-softness protein, friabilin. High-performance anion-exchange chromatography and gel-permeation analysis of the hydrolysis products indicated the preferential hydrolysis of highly branched structures by the enzyme; wheat arabinoxylan and rye (Secale cereale) arabinoxylan (high arabinose to xylose ratios) were hydrolyzed more efficiently by this enzyme than oat (Avena sativa) spelt xylan (low arabinose to xylose ratios). The release of the hydrolysis products as a function of time suggested that the endoxylanolytic activity was associated with the release of arabinose units from the polysaccharides, suggesting that the enzyme action is similar to that by endoxylanases from Ceratocystis paradoxa, Aspergillus niger, and Neurospora crassa. Although the enzyme released arabinose from arabinoxylan, it did not hydrolyze p-nitrophenyl-alpha-L-arabinofuranoside. From the above, it follows that the enzyme, called arabinoxylanase, differs from most microbial endoxylanases and from an endoxylanase purified earlier from wheat flour. PMID- 9414566 TI - Adenosine 5'-triphosphate is required for the assembly of 11S seed proglobulins in vitro. AB - Seed protein proglobulins were synthesized from cDNAs in reticulocyte lysates. Most proglobulins were recovered as trimers when translation rates were low, but mostly monomers were recovered at high translation rates. The prevalence of monomers was accompanied by elevated amounts of insoluble protein recovered at the bottom of sucrose density gradients. Apyrase treatment of translation mixtures after synthesis, but before significant assembly occurred, drastically reduced trimer assembly and increased the proportion of insoluble aggregate. These observations indicated that ATP is required for protein folding and/or trimer assembly. The appearance of insoluble aggregated protein when rates of synthesis were elevated or when ATP was absent suggested that protein misfolding had occurred. Trimer assembly was stimulated when wheat germ translation mixtures defective in supporting efficient trimer assembly were supplemented with fractions isolated from endoplasmic reticula of developing pea (Pisum sativum) seeds. Molecular chaperones are likely involved in folding and/or assembly of proglobulin trimers both in reticulocyte lysates and in seeds. Consistent with this hypothesis, trimer formation was reduced when carboxymethylated bovine albumin and alpha-casein, considered to mimic proteins with extended chain and molten globular conformations and thereby compete for Hsp70- and Hsp60-type molecular chaperones, respectively, were introduced into translation mixtures. PMID- 9414567 TI - Isolation and reconstitution of cytochrome P450ox and in vitro reconstitution of the entire biosynthetic pathway of the cyanogenic glucoside dhurrin from sorghum. AB - A cytochrome P450, designated P450ox, that catalyzes the conversion of (Z)-p hydroxyphenylacetaldoxime (oxime) to p-hydroxymandelonitrile in the biosynthesis of the cyanogenic glucoside beta-D-glucopyranosyloxy-(S)-p-hydroxymandelonitrile (dhurrin), has been isolated from microsomes prepared from etiolated seedlings of sorghum (Sorghum bicolor L. Moench). P450ox was solubilized using nonionic detergents, and isolated by ion-exchange chromatography, Triton X-114 phase partitioning, and dye-column chromatography. P450ox has an apparent molecular mass of 55 kD, its N-terminal amino acid sequence is -ATTATPQLLGGSVP, and it contains the internal sequence MDRLVADLDRAAA. Reconstitution of P450ox with NADPH P450 oxidoreductase in micelles of L-alpha-dilauroyl phosphatidylcholine identified P450ox as a multifunctional P450 catalyzing dehydration of (Z)-oxime to p-hydroxyphenylaceto-nitrile (nitrile) and C-hydroxylation of p hydroxyphenylacetonitrile to nitrile. P450ox is extremely labile compared with the P450s previously isolated from sorghum. When P450ox is reconstituted in the presence of a soluble uridine diphosphate glucose glucosyltransferase, oxime is converted to dhurrin. In vitro reconstitution of the entire dhurrin biosynthetic pathway from tyrosine was accomplished by the insertion of CYP79 (tyrosine N hydroxylase), P450ox, and NADPH-P450 oxidoreductase in lipid micelles in the presence of uridine diphosphate glucose glucosyltransferase. The catalysis of the conversion of Tyr into nitrile by two multifunctional P450s explains why all intermediates in this pathway except (Z)-oxime are channeled. PMID- 9414568 TI - Induction of microbial genes for pathogenesis and symbiosis by chemicals from root border cells. AB - Reporter strains of soil-borne bacteria were used to test the hypothesis that chemicals released by root border cells can influence the expression of bacterial genes required for the establishment of plant-microbe associations. Promoters from genes known to be activated by plant factors included virE, required for Agrobacterium tumefaciens pathogenesis, and common nod genes from Rhizobium leguminosarum bv viciae and Rhizobium meliloti, required for nodulation of pea (Pisum sativum) and alfalfa (Medicago sativum), respectively. Also included was phzB, an autoinducible gene encoding the biosynthesis of antibiotics by Pseudomonas aureofaciens. The virE and nod genes were activated to different degrees, depending on the source of border cells, whereas phzB activity remained unaffected. The homologous interaction between R. leguminosarum bv viciae and its host, pea, was examined in detail. Nod gene induction by border cells was dosage dependent and responsive to environmental signals. The highest levels of gene induction by pea (but not alfalfa) border cells occurred at low temperatures, when little or no bacterial growth was detected. Detached border cells cultured in distilled water exhibited increased nod gene induction (ini) in response to signals from R. leguminosarum bv viciae. PMID- 9414569 TI - The PsaD subunit of photosystem I. Mutations in the basic domain reduce the level of PsaD in the membranes. AB - The PsaD subunit of photosystem I (PSI) is a peripheral protein that provides a docking site for ferredoxin and interacts with the PsaB, PsaC, and PsaL subunits of PSI. We used site-directed mutagenesis to determine the function of a basic region in PsaD of the cyanobacterium Synechocystis sp. PCC 6803. We generated five mutant strains in which one or more charged residues were altered. Western blotting showed that replacement of lysine (Lys)-74 with glutamine or glutamic acid led to a substantial decrease in the level of PsaD in the membranes. The mutant PSI complexes showed reduced NADP+ photoreduction activity mediated by ferredoxin; the decrease in activity correlated with the reduced level of PsaD. Using protein synthesis inhibitors we showed that the degradation rates of the mutant and wild-type PsaD were similar, indicating a defect in the assembly of the mutant protein. Treatment of the mutant PSI complexes with a different concentration of NaI showed that the mutations decreased affinity between PsaD and the transmembrane components of PSI. With glutaraldehyde, the mutant and wild type PsaD proteins could be cross-linked with PsaC, but the PsaD-PsaL cross linked product was reduced drastically when arginine-72, Lys-74, and Lys-76 were mutated simultaneously. These studies demonstrate that the basic residues in the central region of PsaD, especially Lys-74, are crucial in the assembly of PsaD into the PSI complex. PMID- 9414571 TI - The electronic Plant Gene Register. PMID- 9414570 TI - Oxidative stress causes ferredoxin-NADP+ reductase solubilization from the thylakoid membranes in methyl viologen-treated plants. AB - The flavoenzyme ferredoxin-NADP+ reductase (FNR) is a member of the cellular defense barrier against oxidative damage in Escherichia coli. We evaluated the responses of chloroplast FNR to methyl viologen, a superoxide radical propagator, in wheat (Triticum aestivum L.) plants and chloroplasts. Treatments with the herbicide showed little effect on the levels of FNR protein or transcripts, indicating that expression of this reductase is not upregulated by oxidants in plants. Viologens and peroxides caused solubilization of active FNR from the thylakoids into the stroma, converting the enzyme from a membrane-bound NADPH producer to a soluble NADPH consumer. This response appeared specific for FNR, since other thylakoid proteins were unaffected by the treatments. The reductase binding protein was released together with FNR, suggesting that it might be the target of oxidative modification. Stromal accumulation of a functional NADPH reductase in response to oxidative stress is formally analogous to the induction of FNR synthesis observed in E. coli under similar conditions. FNR solubilization may be playing a crucial role in maintaining the NADPH/NADP+ homeostasis of the stressed plastid. The unchecked accumulation of NADPH might otherwise increase the risks of oxidative damage through a rise in the Mehler reaction rates and/or the production of hydroxyl radicals. PMID- 9414572 TI - [Chromosomal aberrations induced in human spermatozoa by mitomycin C]. AB - Human spermatozoa were exposed to Mitomycin C (MM() at the concentrations of 7.5, 15, 3O micrograms/ml for 1 hour respectively before interspecific in vitro fertilization with zona-free hamster ova. The first cleavage metaphases was analyzed to investigate MMC-induced effects on human spermatozoa. The results showed a distinct dose-response relationship in the induction of chromosomal aberrations. The chromatid aberrations were the predominant types in MMC-induced changes. Some chromosome-type aberrations were also observed. This suggested that the induced effects of MMC on human spermatozoa were some different from that of ultraviolet light-like clastogens on somatic cells. 36.89% of MMC-induced aberrations were rejoining-type. This indicated that the DNA repair systems present in golden hamster oocytes were effective in repairing MMC-induced DNA lesions in human spermatozoa. PMID- 9414573 TI - [Primary and secondary structure of 3'--end of the large subunit ribosomal RNA of silkworm Attacus ricini and evolution implications as inferred from the gene sequence]. AB - The DNA sequence of the 3'-end of LSu-rRNA coding region of silkworm Attacus ricini was determined. By comparisons with the corresponding sequence of H. sapiens, X. leavis, H. momus, A. albopictus, D. melanogaster, C. elegants, S. pombe, M. musculus and M. racemosus, we found that the coding sequence we obtained is very conserved. Phylogenetic tree inferred by Neighbor-joining Method showed that the rate of evolution of insects was much faster than vertebrate. In addition, the secondary structure has also worked out it is highly conserved. PMID- 9414574 TI - [Effects of SAD on cytogenetics of mouse embryonic cells and its embryo development]. AB - We report here that SAD may not only suppress transcription activity of rRNA genes, but also increase SCE frequency of mouse embryonic cells, leading to bloching of the embryo development and fetal death, and finally significant decrease of birth percentage of the new born offsprings. Ata concentration of SAD in 10(-5)mol/L, the transcription activity of rRNA genes was decreased to 15% of the normal level, and the SCE frequency was increased doubly, leading to that the birth percentage was decreased to 50% of normal one. When SAD concentration of 10(-3)mol/L was used the transcription activity of rRNA genes was completely suppressed and SCE frequency was increased to as 4.8 times high as normal, leading to that the birth percentage dropped to 4% from 100%. Our results indicates that both the cytogenetic effects and the inhibition of the embryonic cells are directly proportional to the SAD concentration used. The relationship between the change of both the rRNA gene transcription and SCE frequency and the degree of embryo development are also discussed preliminarily. PMID- 9414575 TI - [Study on the recognition and evolutionary genetics of the courtship song of species in Drosphila nasuta species subgroup]. AB - The subgroup of Drosophila nasuta consists of 14 species, subspecies and taxa. It has a wide distribution in area of Indo-Pacific Ocean. The courtship song of species in nasuta subgroup is recorded for the first time in the paper. Some parameters of temporal pattern of pulse song were measured: the inter burst interval (IBI), the interpulse interval (IPI), the length of a pulse train (PTL), the number of pulses in a train (PN), the length of a pulse (PL) and the length of a cycle (CL). Computer analysis techniques were used to make spectral analysis of sine song. A three-dimensional power spectra of the sound was made. In this subgroup, the male of D. pulauna and Taxon-F didn't produce any sound signals during courtship, suggesting that the visual stimuli play an important role in mating process. By analysing the courtship sound signals of other species, subspecies and taxa, the sounds produced by courting male could be described as pulse song or as sine song. By studying the sounds of intra- or interspecific F1 hybrids we have found that the parameters of the temporal pattern of pulse song, for example, the mean value of IPI is controlled by X-linked and autosomal polygene and the sine song frequency is predisposed to the maternity. The phylogenetic tree of the subgroup is constructed based on the temporal pattern of the pulse sounds in different species, subspecies and taxa. The relationship within the members of the Drosophila nasuta subgroup is discussed. PMID- 9414576 TI - [Plasmid DNA fragments from Halobacterium halobium active as eubacteria promoters in Escherichia coli]. AB - In this paper; a promoter-probe plasmid pKK232-8 was used as a vector which functioned in E. coli. The plasmid pHH205 of Halobacterium halobium J7 was digested by two groups of restriction endonucleases BamHI-SalI and HindIII -SalI, recombined in vitro and transformed into E. coli. The transformants were selected on resistance plates to contain ampicillin (Am) and chloramphenicol (Cm). From random-selected 20 strains of transformants we obtained transformants T1 and T2, whose level of Cm resistance got to 110 micrograms/ml. The recombinant plasmids from T1 and T2 were named pJH and pJB, respectively. The resultes of analysis by restriction endonucleases digestion and molecular hybridization showed that recombinant plasmid pJH carried a inserted fragment with size of 800bp from plasmid pHH205. Experimental results of retransfomation proved further that the DNA fragment had promoter function in E. coli. Thus, this indicated that DNA fragments from plasmid of archaebaeteria (H halobium) may function as eubacteria (E. coil) promoters. PMID- 9414577 TI - [Screening for Chlamydia trachomatis?]. PMID- 9414578 TI - [Recent progress on analytical chemistry and biochemistry of D-amino acids]. AB - Recent findings that D-amino acids, especially D-aspartic acid and D-serine, exist in vivo in the mammalian tissues (brain and peripheries), prompted us now to investigate their biological and pathological roles in mammals. In this review, the overview of the progress of analytical chemistry and biochemistry of D-amino acids is described. PMID- 9414579 TI - [Microanalysis of tryptophan metabolites]. AB - In tryptophan metabolites, 3-hydroxykynurenine and 3-hydroxyanthranilic acid have been reported to show a carcinogenic action to mice bladder and the relation of the metabolites to human bladder cancer has been discussed. We developed methods for the fluorometric assay of these compounds and showed that the excretion of 3 hydroxyanthranilic acid increased in the patients with bladder cancer. We also devised methods for the fluorometric assay of glucuronide and sulfate of 3 hydroxyanthranilic acid and showed that the minor excretion of these conjugated forms was shown in humans. The distribution of these compounds was also studied and the obtained data suggests that 3-hydroxykynurenine has affinity for the pancreas. We then developed methods for the determination of other metabolites of tryptophan. A fluorescence reaction with UV irradiation was found and applied to the determination. This method is the most sensitive to kynurenic acid but can be applied to kynurenine, nicotinamide and quinolinic acid. Furthermore, this methods also applied to the determination of some medicines, e.g. indomethacin, isoniazid, nalidixic acid, nicorandil and disodium cromoglicate in the serum or urine. We further devised other methods for the determination of xanthurenic acid and 5-hydroxyindoles. PMID- 9414580 TI - [Selenium methylation and toxicity mechanism of selenocystine]. AB - Selenium is an essential trace element and a toxicant for animals. Selenocystine, a selenium-containing amino acid, is one of the chemical forms in which selenium exists in food. This review summarized recent studies on the toxicity mechanism of selenocystine in experimental animals. Hepatotoxicity is caused by repeated oral administration of selenocystine. Selenocystine is metabolized by reduced glutathione and/or glutathione reductase to hydrogen selenide via selenocysteine glutathione selenenyl sulfide. The hydrogen selenide is a key intermediate in the selenium methylation metabolism of inorganic and organic selenium compounds. Accumulation of the hydrogen selenide resulting from inhibition of the selenium methylation metabolism, detoxification metabolic pathway of selenium, is found in animals following repeated administration of a toxic dose of selenocystine. The excess of the hydrogen selenide produced by inhibition of the selenium methylation metabolism contributes to the hepatotoxicity caused by selenocystine. PMID- 9414582 TI - [Analysis of neurosteroids]. AB - The term neurosteroids applies to those steroids that are both synthesized in the nervous system, either de novo from cholesterol or from steroid hormone precursors, and that accumulate in the nervous system to levels that are at least in part independent of steroidogenic gland secretion rates. Neurosteroids consist of 17- or 20-oxosteroids and accumulate in the brain as unconjugated form and their sulfates, fatty acid esters and sulfolipids. The characterization and determination of neurosteroids including conjugates in the brain are summarized in this review. For example, the separation and characterization of 3-fatty acid esters (stearate, palmitate) of pregnenolone and dehydroepiandrosterone in the rat brain are carried out using liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (LC/APCI-MS) operating in the positive-ion mode. The fatty acid esters obtained from the rat brain were derivatized with O methylhydroxylamine to give the respective methyloximes, which were identified in comparison with their chromatographic behavior with authentic samples during LC/APCI-MS. The function of the steroids are also briefly described. PMID- 9414581 TI - [Chronic renal failure and guanidino compounds]. AB - Guanidino compounds are known as uremic toxins which increase in the blood of patients with renal failure. Guanidino succinic acid (GSA) and methyl guanidine (MG) have been intensively studied since they are toxic and are candidate markers which reflect the pathological stage of nephritis. GSA correlates well with blood urea nitrogen and therefore indicates the reduction of renal function. MG does not appear in the early stage of renal failure and abruptly increases at the stage of serious uremia. MG is produced by the oxidation of creatinine (CTN) with active oxygen. The MG/CTN ratio in the serum therefore reflects the degree of the generation of active oxygen. Accordingly, active oxygen scavengers may be useful for the treatment of uremia. PMID- 9414583 TI - [Methodological approach to regulation of polyamine]. AB - This brief article is dedicated to the late Professor Morizo Ishidate, and concerned in a methodology developed by the author and collaborators, aiming at the regulation of polyamine, especially, the regulation of spermidine synthase. The content is separated in 3 sections. The first section on the development of analytical methods, contains seven items, e.g. fluorometric determination of polyamines by fluorescamine, analysis of naturally occurring polyamines by GC and GC-MS, etc. The second section on the syntheses of needed compounds, contains three items, e.g. syntheses of decarboxylated S-adenosylmethionine which is a substrate for spermidine synthase and its related compounds, syntheses of 15N enriched polyamines applicable to the preparation of various polyamines, etc. The last section on the studies of aminopropyl transferases, contains three items, e.g. purification of spermidine synthase from mammalian tissues using ATPA Sepharose, a novel affinity carrier, newly developed inhibitors for the enzymes, etc. PMID- 9414584 TI - [Diversity of joro spider toxins]. AB - In a study to solve a mystery of venom toxicity of the joro spider, Nephila clavata, we purified and identified novel spider toxins such as clavamine, spidamine and joramine. Chemical analyses, bioassays and physical analyses were specifically elaborated in these procedures. The structure-activity relationship of the spider toxins was discussed biologically and chemically in comparison with the other spider toxins. We considered that the diversity of the joro spider toxins by reserving 2,4-dihydroxyphenylacetyl-L-asparaginylcadaverine as a common moiety gave rise to an important insight into not only the toxic reaction, but also the ontology. PMID- 9414585 TI - [Characterization of spider venom by mass spectrometry, construction of analytical system]. AB - Spiders belonging to the genera, Nephila, Nephilengys, Araneus, etc., possess various polyamine toxins in their venom glands which paralyze insects by blocking the nerve-muscle signal transduction of glutaminergic synapses. More than 50 kinds of polyamine toxin analogs have been characterized which consist, in general, of the aromatic or heteroaromatic moiety connected with the combination of various types of polyamine chains. We are developing a new analytical system by means of a modern mass spectrometric technique which satisfies more rapid, highly sensitive and simple clean-up procedure without complicated chromatographic treatment. The trial of such approach has been performed by employing crude spider venom as the model material. This review article concerns with such proceeding of mass spectrometric analysis by microcolumn HPLC hyphenated FAB-MS system, continuous flow FAB-MS/MS system with high energy collision charge-remote fragmentation, solid and liquid matrix assisted laser desorption ionization (MALDI- and liquid MALDI-) technique connected with tandem mass spectrometer performing in the institute and in addition, femto-molar characterization of new spider venom spider polyamine toxins as a result of the application of such new techniques is addressed. PMID- 9414586 TI - [Multiplicity of sulfotransferases]. AB - Sulfation is an important conjugation reaction in the metabolism of a diversity of xenobiotics and endogenous compounds such as drugs, food additives, carcinogens, steroids and neurotransmitters. Sulfate conjugation is catalyzed by various kinds of sulfotransferase (ST) such as hydroxysteroid ST (HS-ST), phenol ST (P-ST) and estrogen ST (E-ST) present in cytosols. Our laboratory has been studying the multiplicity of rat and mouse STs. We found that tertiary amines such as triethylamine selectively inhibited rat hepatic HS-ST. Developmental changes and zonal distributions of rat liver HS-ST and P-ST isoenzymes provided evidence for their complex regulatory mechanisms. Studies on site-directed mutagenesis and chimeras of rat HS-ST cDNAs, ST-40 and ST-20, revealed the importance of the C-terminal region for the substrate specificity and involvement of multiple regions for the enzyme activities. These two cDNAs have been mapped to the same chromosomal region 1q21.3-->q22.1 by fluorescence in situ hybridization. Mouse olfactory P-ST was present in the cytoplasm of olfactory sustentacular cells and its cloning study revealed that it is 94% identical with rat ST1C1 in amino acid sequences. PMID- 9414587 TI - [Regulation of biological functions by the Kallikrein-Kinin system]. AB - Research on the kallikrein-kinin system started from the discovery of urinary hypotensive substance in Germany around 1940-1950. Since then, numbers of researchers have explored this field including related inhibitors, enzymes and autacoids, from all over the world. Components of the kallikrein-kinin system have been analysed extensively, especially since the epoch of the discovery of the deficient patients in these components. Recent progress of gene techniques also enhanced the progress of the study in the kallikrein-kinin field. In this review I discuss about 1) Components and the difference of plasma kallikrein system and glandular kallikrein system; 2) Impact of the discovery of the deficient plasma in Factor XII, prekallikrein, kininogens as well as in C1INH, and consequent knowledge from the studies of these deficients; 3) Biological roles of the kallikrein-kinin system; and recent topics in this field. PMID- 9414588 TI - [Mechanism of decoding mRNA in protein biosynthesis]. AB - Various experimental data have been supporting an idea that the conformation of A site tRNA is different from that of P-site tRNA and have led to a new tRNA docking pair model, in which the highly conserved G18 and G19 of D-loop in A-site tRNA and C56 and C61 of T psi C-loop in P-site tRNA base pair exist along with the conventional base pairs of adjacent codon-anticodon interactions. This A-P tRNA pair model can be translocated to the P-E tRNA model without changing the conformation except the ACCA termini, keeping the position of the growing nascent polypeptide chain. On the other hand, it is noteworthy that C1378 of E. coli 16S rRNA cross-links to the 32 position on the anticodon loop of A-site tRNA in the pre-translocational state, and also to the same position of E-site tRNA in the post-translocational state, instead of the corresponding position of P-site tRNA. It resulted in a relationship between the A-P and P-E tRNA docking pair models in the pre- and post-translocational states, respectively, caused by a rotation with the angle of 25 degrees around the axis of rotation symmetry. Furthermore, nucleotide sequence analysis showed that CGAGC1107 of 16S rRNA is complementary with the conserved GT psi CG57 of tRNA. When it is combined with the P-E tRNA pair model, the crystallographically obtained L-shaped tRNA model fits both A site codon with base pairings and the free T psi C-loop of P-site tRNA without base pairings. The base pairings between the GT psi CG57 of tRNA and the CGAGC1107 of 16S rRNA destabilize the bound aminoacyl-tRNA and result in a flow of discarding noncognate and near-cognate ternary complexes until cognate one arrives at the A-site codon. Recognition of a cognate ternary complex could occur, starting from breaking a hydrogen bond between N3 atom of U33 and O5' atom of A36 in the aminoacyl-tRNA, with base pairings of the codon-anticodon interactions, the conserved A 1394 in the 16S rRNA to the conserved U33 in the anticodon loop of the tRNA. The exposed U33 of the aminoacyl-tRNA is paired with A1394 in the recognition mode of A site, and finally passed to A1398 of A-site tRNA in the A-P tRNA pair model of the pre-translocational state. The exposure of U33 base at the A site is a key event in the mechanism of codon recognition. PMID- 9414589 TI - [Differences between phthaleins and sulfonphthaleins]. AB - Differences in color and molecular structure between phthalein-pigments and sulfonphthalein-pigments were investigated using X-ray crystallography and absorption spectrophotometry of their aqueous solutions. The molecular structure of sulfonefluorescein (H2+SF-) was determined as a zwitter ion, 2-(3-hydroxonio-6 hydroxy-3H-xanthen-9-yl)benzenesulfonate. The absorption spectra of H2+SF- demonstrated the dissociation profile of a dibasic acid, while those of fluorescein (H2FL) indicated a tribasic acid and further, at pH > 10, SF2- and FL2-, while at pH < 0, H2+SF- and H3+FL to be dominant. The spectra of H2+SF- were analyzed to obtain the values of pK1 and pK2 together with the spectrum of HSF-. Similarly, from the spectra of H3+FL the values of pK1, pK2 and pK3 together with the spectra of HFL- and H2FL were obtained. Further by adding 1/5 of the H3+FL spectrum to 2/15 of HFL- spectrum, an indicated spectrum as that of H2FL was obtained. From these results, the features of dissociation of H2SF- and H3+FL were estimated. The molecular structure of phenolsulfonphtalein (H2PS-) was determined as a zwitter-ion, alpha-(4-hydroxonio-2,5-cyclohexadiene-1-ylidene) alpha- (4-hydroxyphenyl)-2-toluenesulfonate. The absorption spectra of H2PS- demonstrated that H2PS-, HPS- and PS2- became dominant at pH << 0, pH = 4 and pH > 11, respectively. On the contrary, phenolphthalein (H2PP) displayed only one type of absorption spectrum in the visual region, the shape of which was similar to that of PS2- while the molar extinction coefficient was smaller. The spectra were analyzed to obtain the values of pK1 = 9.05 and pK2 = 9.50. The spectra also demonstrated a slow addition reaction of OH- to PP2- and pK3 = 12 was obtained by measuring the absorbance after equilibration. From these results, the features of dissociation and coloration of H2+PS- and H2PP were estimated. PMID- 9414590 TI - [Polarizing microscopy of crystalline drugs based on the crystal habit determination for the purpose of a rapid estimation of crystal habits, particle sizes and specific surface areas of small crystals]. AB - In 1939 the author reported the results of measured refractive indices of about a hundred crystalline drugs listed in [JP V] at the Takeda Research Laboratory using a Leitz PM polarizing microscope and newly developed immersion oils. When the author had reopened the study of crystalline drugs using a polarizing microscope at the Kobe-Gakuin University starting from 1975 one of the main purposes was to clarify the relation between crystal habits and refractive indices. It had been found that in most cases of crystal habits refractive indices were uniquely measured from a predominant pair of faces forming superior the habit, and they were called as "key refractive indices". The author and his co-workers tried to investigate the possibility of measuring the key refractive indices widely from all the obtainable crystalline drugs listed in the [JP X] or [JP XI], co-operating with the Pharmacy of Kobe University Hospital. Thus, more than 170 kinds of crystalline drugs were tested for their key refractive indices and found that they were measured from about 60-70% of tested drugs. It was also clarified that the difference of 2 key refractive indices, (n2 - n1), the birefringence of the section, was also an unique invariable number for the habit, and it played an important role not only for the graphic representation of log(n2 - n1), abscissa, against (n1, n2), ordinate, for the sake of an analytical purpose but also to measure a thickness of a section (habit) using a retardation color. Then, it had been cleared that the similarity of crystal habits in the microscopic field was based on the facts of measuring the same key refractive indices, and the author had developed a chart for measuring key refractive indices as well as producing a 3 dimensional orthographic projection of a crystal habit simultaneously applying a thickness measuring method using a birefringence. Finally 3 dimensional parameter a, b, c of a crystal habit and "habit coefficients" T: square root of ab/c and L: b/c were determined from the orthographic projection. In conclusion using the similarity in crystal habits the distributions of particle sizes and specific surface areas of all the crystals in the microscopic field had been calculated by a personal computer putting in necessary habit coefficients and obtained data of parameter b. The relation between 2 dispersions of particle sizes in log (V) and specific surface areas in log (SSA) were shown under the rectangular coordinates log (V) on the abscissa and log (SSA) on the ordinate, where the loci of log (SSA) formed simple striped pattern composed of parallel straight lines depending on habit coefficients. It would be possible to estimate the value of a specific surface area of any crystalline substance by plotting the value of log (V) on the straight line of a locus of log (SSA) having the same habit coefficients. PMID- 9414591 TI - [Coumarin-containing chiral discriminating agents. VII. New crystalline 1H-NMR enantiomeric excess determination reagent for alcohols and amines, (R)-(-)- and (S)-(+)-O-coumarinylmandelic acids]. AB - The Mitsunobu reaction of commercial 4-hydroxycoumarin with both (R)-(-)- and (S) (+)-tert-butyl mandelates derived from commercial enantiopure mandelic acids furnished (S)-(+)- and (R)-(-)-tert-butyl O-coumarinylmandelates which were, then, treated with trifluoroacetic acid to give novel crystalline optically pure (S)-(+)- and (R)-(-)-O-coumarinylmandelic acids [SCMOH and RCMOH], respectively in good overall yields. Diastereotopic nonequivalence 1H-NMR examination of the resultant esters and amides without any racemization or kinetic resolution by way of a Steglich's procedure (a DCC-DMAP method) has proved each acid to be a useful, efficient and reliable chiral derivatizing agent for the enantiomeric excess determination of chiral alcohols and amines. PMID- 9414592 TI - [Asymmetric synthesis using chiral bases]. AB - Studies have been made to design chiral bidentate lithium amides and chiral tetradentate amines, and to explore the use of these chiral bases for enantioselective formation and reactions of lithium enolates. Chiral bidentate lithium amides having a chiral amide nitrogen made by virtue of chelation were successfully applied to the enantioselective deprotonation reaction of prochiral cyclic ketones, the kinetic resolution of racemic cyclohexanone derivatives by deprotonation, and the regioselective deprotonation of optically active 3-keto steroids. Structures of some of these chiral bidentate lithium amides in the solid state and in solution were elucidated by X-ray and NMR spectroscopic analyses. By the use of chiral tetradentate amines, enantioselective reactions of lithium enolates with electrophiles, such as alkylation, protonation, and Michael addition, proceeded successfully. Examples of catalytic enantioselective deprotonation, alkylation, and protonation by the present strategy are also presented and discussed. PMID- 9414593 TI - [Activation of skin cells by inorganic compounds]. AB - It has been known that the stimulation of skin cells by physiologically active substances is accompanied by the quantitative and qualitative alterations of the glycosaminoglycans (GAGs) in skin. This phenomenon make it possible to evaluate the activity of test substances for the stimulation of skin cells. The purpose of our study is to elucidate the effectiveness of inorganic compounds for the stimulation of skin cells. (1) The cultured skin cells including keratinocytes and fibroblasts, (2) the cultured model skin constituted of collagen gel, fibroblasts and keratinocytes, and (3) the mouse damaged skin were used in the present study. The results obtained from these studies demonstrate that inorganic substances commonly have a possibility to activate cyclic AMP-dependent protein kinase (A-kinase), and the A-kinase activation results in an increase in GAGs. These findings brought the scientific basis for the effectiveness of the Japanese traditional balneotherapy for damaged skin. PMID- 9414594 TI - [The deterioration of hot spring resources and the change of chemical constituents with the development of spa]. AB - General aspects of the deterioration and typical examples in some hot springs (Atami, Shirahama, Isawa.Kasugai, and Syuzenji Hot Springs) are outlined in this report in connection with the change of chemical constituents. (1) Deterioration of thermal spring resources generally occurs accompanied with the following phenomena: a) drawdown of the water level in thermal water wells. b) lowering of temperature of thermal springs. c) change of the water quality (salt water encroachment in the coastal region and decrease in concentration of chemical components resulting from the intrusion of underground water in the inland area). (2) Typical examples of the deterioration are summarized as follows; [table: see text] (3) It has been revealed from the analyses of the statistical data of Environment Agency that the deterioration of hot spring resources in Japan is gradually progressing. PMID- 9414595 TI - [The interim report on survey of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans in marine products and estimation of exposure through fishes and shellfishes]. AB - Survey of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans has been carried out for four years starting from 1992 to 1995, which showed that about 25% or more of marine products are contaminated by trace amount of dioxins and related compounds. But intake of dioxins and related compounds through fishes and shellfishes was below 5 pg/kg/d. Further monitoring is indispensable. PMID- 9414596 TI - [Chemiluminescent assay for biological substances using lucigenin]. AB - The chemiluminescent reaction of lucigenin with various biological substances has been studied. Chemiluminescence of lucigenin is produced by the addition of either hydrogen peroxide or organic reducing compounds to lucigenin in an alkaline solution. On the basis of these reaction, we have developed highly sensitive chemiluminescent methods for the detection of enzyme immunoassay, especially using alkaline phosphatase as a label enzyme, and also for HPLC of corticosteroids or p-nitrophenacyl esters of carboxylic acids. The detection limits of enzymes were 10(-19)-10(-20) mol per assay, corticosteroids and p nitrophenacyl esters were 500 fmol per injection. PMID- 9414597 TI - [Development of new anticancer drugs--the historical background and current status in Japan]. AB - Historically the development of new anticancer drugs in Japan was initiated in 1950s by M. Ishidate et al. for alkylating agents and by H. Umezawa, T. Hata et al. for antitumor antibiotics. In current cancer chemotherapy, appreciable clinical responses have been reported against some types of hematological and childhood malignancies and solid cancers such as chorionic, ovarian, testicular cancers. However, solid cancers such as many GI-tract, lung, breast and other cancers are still drug-refractory. Since many of these cancers have been discovered at their advanced forms, the drug treatment has been a very important modality. From these reasons, efforts have been placed on the search of new anticancer drugs. Recently, new anticancer drugs relating to taxanes, camptothecins, platinum compounds have exhibited excellent clinical activities against some solid cancers. From these experiences, searches of novel anticancer drugs including those against molecular targets have been continued. The author is greatly indebted to the late Professor Morizo Ishidate for the initiation of his career in cancer research. PMID- 9414598 TI - [Structure and metabolic fate of N-nitrosodialkylamines in relation to their organotropic carcinogenicity with special reference to induction of urinary bladder tumors]. AB - The metabolic fates of N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN) and N,N dibutylnitrosamine (DBN) were investigated in the rat and other animal species, to elucidate a possible relationship between metabolism and organotropic carcinogenicity to the urinary bladder of these N-nitrosamines. The principal urinary metabolite of BBN as well as of DBN in the rat was N-butyl-N-(3 carboxypropyl)nitrosamine (BCPN), which was demonstrated to be the active form of these compounds as bladder carcinogen. The species difference in response to BBN or DBN is discussed on the basis of the urinary excretion rate of BCPN. Metabolism in vivo and carcinogenicity of a number of BBN analogues were investigated in the rat and a general scheme for biotransformation of N-alkyl-N (omega-hydroxyalkyl)nitrosamines is given. A possible correlation of structure and metabolism with organotropic carcinogenicity of BBN analogues is discussed, with special reference to selective induction of urinary bladder tumors. PMID- 9414599 TI - [Genetic polymorphisms in xenobiotic metabolizing enzymes as a determinant of susceptibility to environmental mutagens and carcinogens in humans]. AB - Xenobiotic metabolizing enzymes are known to play a role in the metabolic activation of environmental mutagens and carcinogens to exert their carcinogenic effects as well as detoxification by increasing their hydrophilicity. These enzymes include cytochrome P450s, glutathione S-transferases (GSTs), acetyltransferases (NATs) and sulfotransferases. Genetic polymorphisms in many of these enzymes, such as CYP1A1, CYP1A2, CYP2C9, CYP2D6, CYP2E1, NAT1, NAT2, GSTM1, GSTP1 and GSTT1, have been shown to occur, which result in the altered expression of enzymatic activities. This suggests that the genetic polymorphisms may affect the individual susceptibility to environmental carcinogens and thus play a role in human carcinogenesis. Recently, the mutations that confer those polymorphisms of xenobiotic metabolizing enzymes have been identified and genotyping methods for the genetic polymorphisms have been developed. Specific phenotypes and genotypes for CYP1A1, CYP2D6, CYP2E1, NAT1, NAT2, GSTM1 and GSTP1 have been associated with susceptibility to malignant diseases including lung, bladder and colon cancers, although the association was not confirmed in some studies. A number of factors such as degree of exposure to environmental carcinogens and the role of xenobiotic metabolizing enzymes in human carcinogenesis should carefully be evaluated in understanding genetic susceptibility. PMID- 9414600 TI - [Lethal drug interactions of the new antiviral, sorivudine, with anticancer prodrugs of 5-fluorouracil]. AB - In 1993 eighteen Japanese patients with cancer and herpes zoster, a viral disease, died from interactions of the new oral antiviral drug, sorivudine (SRV: 1-beta-D-arabinofuranosyl-(E)-5-(2-bromovinyl)uracil), with oral anticancer prodrugs of 5-fluorouracil (5-FU) within 40 d after SRV was approved by the Japanese government and began to be used clinically. Before the death, most of these patients had severe symptoms of toxicity, including diarrhea with bloody flux and marked decreases in white blood cell and platelet counts. All of these patients received SRV daily for several days while being administered long-term anticancer chemotherapy with one of the oral 5-FU prodrugs. There was no acute toxic symptom in patients who received SRV alone or SRV and the other types of anticancer drugs. A toxicokinetic study was carried out using rats to investigate the mechanism of the acute death in the patients caused by drug interactions between SRV and 5-FU prodrugs. Rats were orally coadministered SRV with tegafur (FT: 1-(2-tetrahydrofuryl)-5-fluorouracil), a 5-FU prodrug that most of the patients were considered to receive before the death. All the rats receiving SRV and FT once daily showed extremely elevated levels of 5-FU in the plasma and tissues, including bone marrow and small intestines, and died within 10 d, while the animals given the same repeated dose of SRV or FT alone were still alive over 20 d without any appreciable toxic symptom. Before their death, there were a marked damage of bone marrow, a marked atrophy of intestinal membrane mucosa, marked decreases in white blood cells and platelets, diarrhea with bloody flux, and severe anorexia as reported for the patients. Data obtained by in vivo and in vitro studies indicated that (E)-5-(2-bromovinyl)uracil (BVU), generated from SRV by the gut flora and absorbed through the intestinal membrane, was reduced in the presence of NADPH to a reactive form by hepatic dihydropyrimidine dehydrogenase (DPD), a key enzyme regulating the tissue 5-FU levels from FT, bound covalently to DPD as a suicide inhibitor, and markedly retarded the catabolism of 5-FU. An irreversible inactivation by BVU of rat and human DPDs, expressed in E. coli for the latter, was observed in the presence of NADPH with their purified preparations in a manner reciprocal to radiolabelling of the enzyme proteins with [14C]BVU. SRV showed no inhibitory effect on the rat and human DPDs in the presence of NADPH. PMID- 9414601 TI - [Doping test for racehorses in Japan]. AB - The doping test method used in a horse race requires the accurate detection of a wide variety of drugs and metabolites as well as the rapidity in order to examine a large number of samples within a limited time. For this purpose, the routine method consists of a preliminary screening and a confirmatory test. In this paper, a historical review for the development of the doping test method in Japan is described. The metabolism and pharmacology of drugs in horses are also discussed. PMID- 9414602 TI - [SMON and pharmacokinetics of chinoform with special reference to animal species difference]. AB - The experimental reproduction of SMON using several kinds of animals given a prolonged administration of chinoform has been carried out by many investigators because of the importance to solve the problem of etiology in the SMON. In these experiments, it is demonstrated that a marked species difference was observed in the relationship between the doses given to animals and the frequency of the onset of neurologic symptoms. Although dogs were accepted as the most suitable animal model for SMON, pathological changes in the peripheral nerve of the dog were not observed. The blood level or tissue distribution of chinoform after oral, intravenous or intraperitoneal administration of the drug differed in the animal species. Thus, it is considered that the species difference in the onset of neurologic symptoms is principally caused by the difference in pharmacokinetics of chinoform in each animal. Moreover, for the onset of neurologic symptoms in animals, perhaps it is necessary to maintain the level of unconjugated chinoform in the nerve tissues around several to over ten micrograms/ml for three or four weeks as well as that in SMON patients while the neurologic symptoms or pathological changes do not appear in some kinds of animals at these levels. In a study on the cellular toxicity of chinoform, many other problems remain to be solved although degeneration or uncoupling on oxidative phosphorylation in mitochondria of the axons by chinoform and lipid peroxidation of the membrane by chinoform-ferric chelate have been already shown. PMID- 9414603 TI - [Synthesis of promin in Japan and global elimination of Hansen's disease]. AB - Prof. Morizo Ishidate synthesized "Promin" for the treatment of leprosy/Hansen's disease which had been considered "incurable" until the discovery of antileprosy effect of that drug by Dr. Faget of U.S.A. in 1941. Prof. Ishidate was the first to synthesize the drug in Japan in 1946 based on a brief news item on a Swiss journal smuggled in during the War. For this achievement, he is known as "father of chemotherapy for leprosy in Japan." Prof. Ishidate also contributed to the global fight against leprosy as the Chairman of the Board of Directors of Sasakawa Memorial Health Foundation, which he helped to establish in May 1974, with a full financial backing of Mr. Ryoichi Sasakawa, President of Japan Shipbuilding Industry Foundation. Prof. Ishidate, with his scientific knowledge as well as christianity based humanitarian concern, advised Mr. Sasakawa how to spend JSIF money wisely for eliminating leprosy and nearly US$200 million was channeled through WHO and SMHF. The successful outcome of global multidrug therapy (MDT) programme in the '80s resulted in the adoptation of resolution by the World Health Assembly, "Elimination of Leprosy, as a public health problem" by the Year 2000. The synthesis of "Promin" in Japan and promoting the global implementation of MDT, both achievement can be attributed to Prof. Ishidate. PMID- 9414604 TI - [New drug development by innovative drug administration--"change" in pharmaceutical field]. AB - New drug development can be made by providing products of higher "selectivity for the drug" for medical treatment. There are two ways for the approach to get higher "selectivity of drug": 1) discovery of new compounds with high selectivity of drug; 2) innovation of new drug administration, that is new formulation and/or method with high selectivity of drug by integration and harmonization of various hard/soft technologies. An extensive increase of biological information and advancement of surrounding science and technology may modify the situation as the latter overcomes the former in the 21 century. As the science and technology in the 21 century is said to be formed on "3H", that is, 1. hybrid; 2. hi-quality; 3. husbandry, the new drug development by innovative drug administration is exactly based on the science and technology of 3H. Its characteristic points are interdisciplinary/interfusion, international, of philosophy/ethics, and systems of hard/hard/heart. From these points of view, not only the advance of unit technology but also a revolution in thinking way should be "must" subjects. To organize this type of research well, a total research activity such as ROR (research on research) might take an important and efficient role. Here the key words are the "Optimization technology" and "Change in Pharmaceutical Fields." As some examples of new drug innovation, our trials on several topical mucosal adhesive dosage forms and parenteral administration of peptide drugs such as insulin and erythropoietin will be described. PMID- 9414605 TI - Why psychologists should treat alcohol and drug problems. AB - Because of the prevalence of substance abuse in general clinical populations, it is important for psychologists to have knowledge and skill in this area. Psychologists also have special expertise to offer in the assessment and treatment of alcohol/drug problems. Current evidence indicates that (a) alcohol/drug problems generally obey ordinary behavioral principles and processes, (b) substance abuse frequently occurs within a broader cluster of psychological problems, (c) the treatment approaches most strongly supported by outcome research are fundamentally psychological in nature, (d) cognitive behavioral principles are of demonstrable value in motivating change in alcohol/drug use, and (e) clinical skills and styles (e.g., empathy) commonly included in the training of psychologists are important determinants of favorable treatment outcomes with substance use disorders. These factors in the context of changing health care indicate that psychologists should play an increasing role in assessing and treating addictive behaviors. PMID- 9414606 TI - Beyond pleasure and pain. AB - People approach pleasure and avoid pain. To discover the true nature of approach avoidance motivation, psychologists need to move beyond this hedonic principle to the principles that underlie the different ways that it operates. One such principle is regulatory focus, which distinguishes self-regulation with a promotion focus (accomplishments and aspirations) from self-regulation with a prevention focus (safety and responsibilities). This principle is used to reconsider the fundamental nature of approach-avoidance, expectancy-value relations, and emotional and evaluative sensitivities. Both types of regulatory focus are applied to phenomena that have been treated in terms of either promotion (e.g., well-being) or prevention (e.g., cognitive dissonance). Then, regulatory focus is distinguished from regulatory anticipation and regulatory reference, 2 other principles underlying the different ways that people approach pleasure and avoid pain. PMID- 9414607 TI - Sex differences in social behavior: comparing social role theory and evolutionary psychology. PMID- 9414608 TI - On the origins of sex differences in social behavior: Darwinian and non-Darwinian accounts. PMID- 9414609 TI - The U.N. Convention on the Rights of the Child: lost in the clash of adverse opinions. PMID- 9414610 TI - Armed conflict and children's rights. PMID- 9414611 TI - Interactions between dansyl amino acids and human serum albumin using high performance liquid chromatography: mobile-phase pH and temperature considerations. AB - The reversed-phase liquid chromatography (RPLC) retention mechanism of a series of dansyl amino acids was investigated over a wide range of mobile-phase pH and column temperatures using human serum albumin (HSA) as a chiral stationary phase. Thermodynamic constants for the transfer of a solute from the mobile to the HSA stationary phases were determined. Different van't Hoff plot shapes were observed with different mobile-phase pH values, indicating a change in the retention mechanism. Enthalpy-entropy compensation revealed that the solute retention mechanism was independent of the compound molecular structure, the same at four pH values (5.5, 6, 6.5, and 8), but changed at pH = 7 and 7.5. Differential scanning calorimetry was used to show phase transition in the HSA stationary phase at pH = 7 and 7.5. A new theory was presented to explain that the HSA protein structure balance between a disordered and an ordered solid-like state. Variations of column temperature and mobile-phase pH tend to cause this phase transition between these two states, explaining the observed thermodynamic constant variations with pH and temperature. PMID- 9414613 TI - Synthesis and characterization of insulin-fluorescein derivatives for bioanalytical applications. AB - Human insulin was labeled with fluorescein isothiocyanate (FITC) and fully characterized to yield four distinct insulin-FITC species. High-performance liquid chromatography and electrospray mass spectrometry were used to determine the extent and location of fluorescein conjugation. By changing the reaction conditions (i.e., pH, time, and FITC/insulin ratio) the selectivity of the fluorescein conjugation was altered, and all conjugates could be separated. The isolated species of insulin-FITC were labeled at the following residues: A1(Gly), B1(Phe), A1(Gly)B1(Phe), and A1(Gly)B1(Phe)B29(Lys). All four insulin-FITC conjugates were then used to develop fluorescence polarization binding assays with monoclonal and polyclonal anti-insulin antibodies. The assay sensitivity differed between the conjugates depending on the site of modification (B1 > A1 > A1B1 > A1B1B29). Also, the type of antibody used had an important role in the binding of insulin-FITC conjugates. Finally, for the first time the biological activity of the four conjugates was demonstrated by an autophosphorylation assay. The positional substitution dramatically affected the biological activity, confirming insights into the residues responsible for the insulin binding region. The B1 conjugate was found to retain almost all biological activity while the A1 and A1B1 conjugates had approximately 10 times lower activity. The trisubstituted species (labeled at A1, B1, and B29) was determined to be least active. PMID- 9414612 TI - High-performance liquid chromatographic analysis of complex N-linked glycans derivatized with 2-aminoacridone. AB - 2-Aminoacridone (2-AMAC) has been used to derivatize mixtures of N-linked oligosaccharides released from alpha(1)-acid glycoprotein and immunoglobulin G. In each case, the HPLC profile obtained for the derivatized glycans was compared to that obtained after digestion with sialidase and a two-enzyme array system made up of sialidase and alpha-fucosidase, prior to derivatization by 2-AMAC. These studies are rapid and provide a wealth of preliminary information about the degree of sialylation and core fucosylation in the corresponding parent glycans. Moreover, collection of glycans from one single injection has provided enough material for molecular weight determination by MALDI-MS analysis. In this study we have also carried out limited MS-MS studies on enriched fractions of 2-AMAC glycans using a nanospray orthogonal quadrupole time-of-flight mass spectrometer. PMID- 9414614 TI - In vivo determination of ultratrace amounts of prostaglandin in plasma by high performance liquid chromatography/laser-induced fluorometry/ultrasensitive laser spectrometry under severe conditions. AB - We succeeded in determining ultratrace prostaglandin amounts in plasma, at the femtomolar level, using laser-induced fluorometry through a complete redesign of the analytical procedures. Practical samples, especially plasma, contain large amounts of admixtures, and prostaglandin in plasma (pg/mL) has been considered to be difficult to detect because the samples and reagents supplied by conventional procedures are neither pure nor stable enough to get good results by ultrasensitive laser spectrometry. We completely redesigned the analytical procedures after careful investigations of the reagent purification and the column separation conditions based on a newly found behavior of the reagent and derivatized prostaglandin in a small quantity of ethanol in the mobile phase. A lower determination limit of 23 pg/mL (65 fmol) was achieved, the variance was 12% at 25 pg/mL, and the recovery rate was 88-89%. This method was applied to in vivo analysis of the concentration of prostaglandin E1 administered as a prodrug of prostaglandin E1 (delta(8)-9-O-butyryl prostaglandin F1 butyl ester, AS-013) by intravenous infusion to beagle dogs. A clear correlation between the change of blood pressure and the prostaglandin E1 concentration was confirmed. PMID- 9414615 TI - Diastereomeric selectivity of carbon-coated zirconia reversed-phase liquid chromatographic media. AB - The determination of enantiomeric excess, that is, the relative amount of any pair of optical antipodes, constitutes a integral part of the work of analytical and synthetic chemists involved in natural products research or pharmaceutical development. Mosher's reagent [alpha-methoxy-alpha-(trifluoromethyl)phenylacetyl chloride] has evolved into a major tool for the determination of absolute configuration by NMR. We report here on the separation of diastereomers formed by derivatizing enantiomers with Mosher's reagent. We have shown that reversed-phase liquid-solid adsorption chromatography on carbon surfaces frequently gives considerably superior resolution of diastereomeric pairs than does RPLC on conventional bonded phases. The improved resolution results from the very high sensitivity of solid carbon surfaces to the geometric organization of the solute rather than from differences in column efficiency. We compare the separation of pharmaceutically and biologically important stereoisomeric mixtures, including (+/-)-warfarin and (+/-)-amino acid esters, on both conventional bonded phases and carbon surfaces prepared by chemical vapor deposition of organic compounds on porous zirconia microparticles. PMID- 9414616 TI - Bioligand interaction assay by flow injection absorptiometry. AB - A novel technique for the study of bioligand interactions based on combining flow injection on renewable surfaces with UV-visible absorptiometry is introduced. The concept is proven by monitoring the binding of various proteins to protein G and the binding of various insulin analogs to a monoclonal anti-insulin antibody, thus providing a comparison with the performance of surface plasmon resonance (SPR)-based techniques. The advantages of the bioligand interaction flow injection absorptiometry approach are speed, low cost, no need for regeneration of solid substrate, and spectral resolution not available with SPR sensing. It is believed that this technique will have an impact on the entire biosensor field since it allows simultaneous monitoring of labeled as well as nonlabeled species in real time over a wide spectral range with high sensitivity. PMID- 9414617 TI - HIV-1 Tat peptide binding to TAR RNA by electrospray ionization mass spectrometry. AB - Electrospray ionization mass spectrometry (ESI-MS) has been used to study the noncovalent complexes formed from the interaction between HIV-1 Tat peptide and Tat protein with TAR RNA. Both positive ion and negative ion ESI mass spectra showed a maximum stoichiometry of 3:1 between Tat peptide and TAR RNA. However, the higher order complexes only occurred at high relative concentrations of Tat peptide. The 1:1 Tat peptide-TAR RNA complex is believed to involve only specific interactions, whereas the higher order complexes involve nonspecific interactions. Relative binding affinities between Tat peptide and TAR RNA and its various mutants (TAR missing the three-nucleotide bulge, TAR with a poly(ethylene glycol) linker in the bulge region, and TAR with a poly(ethylene glycol) linker in the loop region) can be differentiated by competitive binding experiments and ESI-MS measurements. The gas phase mass spectrometry experiments are consistent with solution phase studies, as they show that mutations in the bulge region reduce TAR RNA affinity to Tat peptide. PMID- 9414618 TI - Use of nonvolatile buffers in liquid chromatography/mass spectrometry: advantages of capillary-scale particle beam interfacing. AB - The use of nonvolatile buffers like phosphate and citrate is usually incompatible with mass spectrometric detection or may heavily interfere with ion generation. In the case of conventional HPLC flow rates, the continuous buffer deposition produces fouling of the mass spectrometer ion source and may worsen the performance of any LC/MS interface as well. Our research group demonstrated that reducing the mobile phase flow rate in a particle beam interface improves the nebulization and enhances the overall performance. We have also assumed that such a modification may better handle potentially instrument harmful solvents or nonvolatile HPLC buffers. Since the presence of salts does not interfere with the electron ionization process, the particle beam interface can be considered, in principle, particularly suitable for those chromatographic separations that benefit from nonvolatile buffers. The microscale flow rate interface, developed in our laboratory, generates an aerosol from 1 microL/min of mobile phase flow rate. Under these conditions, the absolute amount of a nonvolatile buffer actually introduced into the system is approximately 1/1000 of that carried by a conventional HPLC column, slowing its deposition. This assumption was verified with a real-world application requiring the use of a phosphate buffer at a maximum concentration of 10 mM. It involved the determination of dexamethasone, an anti-inflammatory drug, in human blood after its administration to a patient. Several crucial mass spectral and chromatographic parameters were monitored during a 35-day period of intense use. Neither appreciable signal modification nor evident buffer deposition was observed during the test period, with only a normal and limited run-by-run and day-by-day variation of the instrument response. PMID- 9414620 TI - Capillary gas chromatography with cryogenic oven temperature for headspace samples: analysis of chloroform or methylene chloride in whole blood. AB - A new and sensitive gas chromatography (GC) method for measurement of chloroform or methylene chloride in whole blood is presented. Trace levels of these analytes present in the headspace of samples were cryogenically trapped prior to on-line GC analysis. After heating of a blood sample containing chloroform and methylene chloride (internal standard, and vice versa) in a 7.0-mL vial at 55 degrees C for 20 min, 5 mL of the headspace vapor was drawn into a glass syringe. All vapor was introduced into an Rtx-Volatiles middle-bore capillary column in the splitless mode at -30 degrees C oven temperature to trap the entire analytes, and the oven temperature was programmed up to 280 degrees C for detection of the compounds and for cleaning of the column. The present conditions gave sharp peaks for both chloroform and methylene chloride and very low background noises for whole blood samples. As much as 11.5 and 20.0% of chloroform and methylene chloride, respectively, which had been added to whole blood in a vial, could be introduced into the GC column. The calibration curves showed linearity in the range of 0.05 5.0 micrograms/0.5 mL of whole blood. The detection limit was estimated to be about 2 ng/0.5 mL. The coefficients of intraday and interday variations were 1.31 and 8.90% for chloroform and 1.37 and 9.03% for methylene chloride, respectively. The data on chloroform or methylene chloride in rat blood after inhalation of each compound were also presented. PMID- 9414619 TI - Monitoring recombinant protein drugs: a study of insulin by H/D exchange and electrospray ionization mass spectrometry. AB - The increasing emergence of new protein- and peptide-based drugs makes necessary the development of rapid and sensitive methods to check consistency between and within batches of biotechnology pharmaceuticals to ensure product quality. We evaluated electrospray ionization mass spectrometry in combination with H/D isotopic exchange as a potential tool, taking as examples for this case study the four insulins used for treating insulin-dependent diabetes. Two (bovine and porcine) are produced naturally, and two are produced by recombinant biotechnology techniques [recombinant human (r-human) and its human insulin analog (LysPro)]. The extent of H/D exchange at a given time was measured with less than 2 micrograms (< 350 pmol) of sample and was sufficient for discriminating among the different insulins. After 60 min, bovine, porcine, r human, and LysPro insulins exchanged on average 25, 28, 30, and 38 amide protons, respectively. After prolonged incubation with D2O for 24 h, bovine and porcine insulins exchanged 31 protons, whereas r-human and LysPro insulins exchanged 34 and 43 amide protons, respectively. The differences in H/D exchange are protein signatures that relate to differences in conformation and folding. The extent of exchange distinguishes among the insulin types and assures the consistency of batch preparations for a given insulin. PMID- 9414621 TI - Quantitative analysis of despropionyl-bezitramide, the active metabolite of bezitramide (Burgodin), in biological samples by high-performance liquid chromatography with fluorescence detection. AB - A sensitive high-performance liquid chromatographic procedure with fluorescence detection was developed for the quantitative determination of despropionyl bezitramide, the active metabolite of bezitramide (Burgodin), in biological samples. Chromatographic separation was achieved on a Hypersil ODS (C18) 5-micron column, using a 31:69 (v/v) mixture of 0.1 M ammonium acetate and methanol/acetonitrile (50:50, v/v)-0.1 M ammonium acetate as the eluent. Internal standardization with N-methyldespropionyl-bezitramide was used in order to enhance the precision and the accuracy of the method. For the isolation of the compound from biological samples, a liquid-liquid extraction with n-hexane isoamyl alcohol (93:7 v/v) was performed. Calibration graphs were prepared for blood and urine, and linearity was achieved over a concentration range 1-50 ng/mL. The quantitation limit for despropionyl-bezitramide in blood and urine was 1 ng/mL. At a 10 ng/mL concentration in blood, coefficients of variation of 3.3 and 6.5% were obtained for within-day and between-day precisions, respectively. For urine, the respective coefficient of variation values of 4.3 and 4.9% were obtained. The selectivity and the accuracy of the method were satisfactory. Samples (blood, urine, stomach contents, bile, liver, kidney) from five fatalities that were due to the combined intake of several drugs, including bezitramide, were analyzed and the results are reported. In addition, one blood sample and 14 urine samples from persons suspected of using bezitramide were analyzed, revealing despropionyl-bezitramide concentrations in urine ranging from 1.3 to 72.3 ng/mL. PMID- 9414623 TI - Ethnolinguistic vitality under a new political dispensation in South Africa. AB - The dimensions and group expectations of ethnolinguistic vitality in the new South Africa were investigated in random samples of 460 Whites (347 Afrikaans speaking; 113 English-speaking) and 466 Blacks. By means of a factor analysis, 5 factors were distinguished: Institutional Support, Group Status and Power, Maintenance of Identity, Maintenance of Symbols, and Threat to Identity. The expectations of groups differed significantly in regard to the dimensions of ethnolinguistic vitality. Relationships between these dimensions and other variables and the implications of the findings were discussed against the background of sociopolitical and economic changes in South Africa. PMID- 9414622 TI - Mismatch-sensitive hybridization detection by peptide nucleic acids immobilized on a quartz crystal microbalance. AB - A quartz crystal microbalance DNA hybridization biosensor, based on thiol derivatized peptide nucleic acid (PNA) probes, offers unusual in situ differentiation of single-base mismatches. A large excess of a single-base mismatch oligonucleotide has no effect on the frequency response of the target. Such remarkable distinction between perfect matches and mismatches is illustrated by the detection of a common mutation in the p53 gene. The greater specificity of the new mass-sensitive indicatorless hybridization device over those of analogous PNA-based carbon electrodes is attributed to the formation of a PNA monolayer and the use of a hydrophilic ethylene glycol linker. The improved specificity is coupled to very fast (3-5 min) hybridization in a low-ionic-strength medium. PMID- 9414624 TI - The influence of societal factors on female body image. AB - This study was designed to identify factors associated with the perceptual and attitudinal components of female body image. The influence of society and factors thought to mediate the relationship between body image and society (field dependence, locus of control, and self-esteem) were investigated. Age and body mass index (BMI) were also included as independent variables. A total of 101 female university students in Australia ranging in age from 18 to 55 years (M = 24.11) participated in the study. A video camera apparatus (VCA) was used to assess perceptual distortion of body size. The VCA, the Body Esteem Scale, and the Appearance Evaluation subscale of the Multidimensional Body Self Relations Questionnaire were used to assess body satisfaction. On average, women underestimated their body sizes by 4%, and they typically wanted to be smaller than their actual body sizes. About two fifths of the women expressed moderate to strong negative feelings about both individual body parts and their bodies as a whole. Multiple regression analyses revealed that perceptual distortion of body size could not be predicted from the independent variables. Body satisfaction was best explained by societal factors, self-esteem, and BMI. PMID- 9414625 TI - Gender as a determinant of work values among university students in Israel. AB - Gender differences in work values, measured by the 25-item Manhardt scale (1972), were examined among 820 (391 male and 429 female) undergraduate students at Ben Gurion University of the Negev, Israel. Male and female students differed on 9 items. The single students' scores were similar to the scores of the total sample; among married students, there were gender differences on only 3 items. In the Faculty of Humanities and Social Sciences, male and female students differed on 13 items; in the Faculty of Engineering, they differed on 8 items; and in the Faculty of Medicine, they differed on 2 items. There was no consistent pattern of gender-based differences in work values. These findings show the need to control for background variables and field of study/occupation in examinations of gender based differences in work values; they also indicate the need to revise and augment traditional explanations of gender differences in work values. PMID- 9414626 TI - Social strategies and loneliness. AB - Although substantial research has been done on loneliness, in only a few studies has the extent of its association with the cognitive and attributional strategies people apply in social situations been investigated. Two studies were carried out among Finnish students to examine this association. In Study 1, 70 men and 202 women filled in the Cartoon-Attribution-Strategy Test (CAST) and Rosenberg's Self Esteem Scale (RSE), then 1 year later, the revised UCLA Loneliness Scale. In Study 2, 25 men and 35 women filled in the CAST and the RSE, then 4 months later, the UCLA Loneliness Scale. In both studies, a pessimistic avoidance strategy was associated with subsequent feelings of loneliness, even after controls for the level of self-esteem. Both an optimistic planning strategy and a self-serving attributional bias were negatively associated with feelings of loneliness among men but not among women. PMID- 9414627 TI - Feelings of mastery in aggressors: a conceptual replication of an American experiment in France. PMID- 9414629 TI - Attitudes of white American male students toward work force diversity programs. PMID- 9414628 TI - Patterns of depression in Xhosa and Yoruba students. PMID- 9414630 TI - Food preservation using ionizing radiation. AB - Irradiation processing has been researched extensively and is now in use worldwide for many food commodities. Irradiation has been successfully used to reduce pathogenic bacteria, eliminate parasites, decrease postharvest sprouting, and extend the shelf life of fresh perishable foods. Although food irradiation is widely accepted in world food markets, U.S. markets have been slower to accept the idea of irradiated food products. For fruits and vegetables, irradiation is not a cure for shelf life problems; cost and quality problems damage preclude its general use. It appears that the most likely use of irradiation in fruits and vegetables is as an insect control in those commodities for which there is no effective alternative method. For grains such as rice and wheat, irradiation has been used primarily to control insect infestation when insects have been shown to develop resistance to the traditional fumigation methods. Treatment of spices with irradiation doses of 10 kGy has proved to extend shelf life without causing significant changes in sensory or chemical quality. Higher doses that effectively sterilize spices, however, may cause undesirable chemical and sensorial changes. For meat, especially red meat, irradiation is considered a viable alternative in the effort to improve the safety of meat products. With time, the authors believe that economic realities and the technical superiority of irradiation for specific poultry products will lead to public acceptance of the process. Irradiation of seafood products is still being considered for approval by the USFDA, although it is currently used in Asian and European markets, especially for shrimp. It is our belief that scientifically based research in food irradiation and the positive results thereof will also prove economical in the twenty-first century. As we move to a more peaceful world with reduced threat of nuclear holocaust, these valid opinions will prevail and will overshadow the distortions and misinformation generated by the opponents of irradiation. PMID- 9414632 TI - The relationship between transience and current life situation in the homeless services-using population. AB - Although transience has been the focus of an enormous amount of public attention, there has been limited empirical research on transience in the homeless population. The purpose of the study discussed in this article was to develop a construct of transience for the homeless services-using population. Transience was defined as consisting of four factors: migration, duration, intention, and involvement. This construct was used to predict current housing and employment status, substance use, receipt of entitlements, and health and mental health services use. The study collected data on 146 individuals. Findings suggest the validity of the study's conceptualization of transience, particularly in its ability to predict current substance use. This study also found indirect evidence for one previously proposed profile of transients--the "transient substance abuser," but this profile may have two categories--used drugs or used alcohol. PMID- 9414631 TI - Cadmium contamination of vegetable crops, farmlands, and irrigation waters. AB - Cadmium, a highly toxic element that can accumulate in living tissues, is a potential threat to the environment and to human health. The main sources of Cd contamination of vegetable crops via farm soils and irrigation waters are reviewed, as are the influence of residual sludges used as fertilizers and the indiscriminate use of pesticides. The principal sources of exposure and toxicological characteristics of Cd are described, together with its effects on human health. Current European technical and health regulations aimed at controlling Cd levels are noted, and the most common analytical techniques for measuring Cd content are summarized. PMID- 9414633 TI - Ideology and social work practice in substance abuse settings. AB - The profession of social work has a unique role in preventing and treating alcohol and other drug problems. In human services settings shared beliefs or ideologies of care are expected to have substantial influence over the way in which problems are perceived and the types of service technologies used. Thus, it is important that social work professionals be cognizant of what beliefs they hold and how their beliefs about substance abuse treatment and prevention may affect practice. This article discusses current ideologies of care in the substance abuse arena, including the disease/abstinence, psychosocial, ecological, and harm-reduction approaches. In addition, this article examines managers' beliefs about substance abuse programs to determine if there are differences between those who have a social work background (that is, hold at least one social work degree) and those who do not. Suggestions for social work practice and future research also are provided. PMID- 9414634 TI - The Index of Drug Involvement: a partial validation. AB - Social workers often need a wider range of assessment scales that can be used in education, research, and practice. The Index of Drug Involvement (IDI) is a short form assessment scale that may be of use to social work educators, researchers, and practitioners who work with clients with drug abuse or chemical dependency problems. This article describes the IDI; provides information about its administration, scoring, and interpretation; and describes the initial research conducted to validate the instrument. This article provides information about the reliability of the IDI; reports the standard error of measurement; and presents findings concerning the content, construct, and criterion validity of the instrument. Also presented is initial information about the development and use of a clinical cutting score that will help practitioners evaluate the clinical significance of a drug abuse problem and that can be a guide for establishing initial and final treatment goals. PMID- 9414635 TI - Immigrant policy: issues for social work practice. AB - Immigrants make up a significant segment of U.S. society. Immigration to the United States has been characterized by steady growth, dramatic changes in ethnic composition, and declining socioeconomic levels. The challenge for social work is to respond to the social services needs of immigrants by designing appropriate programs that will contribute to the social and economic integration of immigrants. This article provides an overview of the major policy issues relevant to social work practice with immigrants and describes the recent U.S. immigrant population. It discusses current federal policy that affects service provision to immigrants, defines immigrant eligibility for social services, outlines the major areas of need among immigrants, and considers implications for social work practice. PMID- 9414636 TI - Sociopolitical antecedents to Stonewall: analysis of the origins of the gay rights movement in the United States. AB - The 1969 Stonewall riot in New York City is widely celebrated as the beginning of the modern gay rights movement. However, the social changes that followed were possible only because of a decades-long indigenous organization. This article examines three traditional organizational theories on social action movements and applies Aldon Morris's analytical framework for understanding collective social action to the emergence of the modern U.S. gay civil rights movement. The author concludes that social workers need to understand how to gain access to and support indigenous social structures to assist oppressed groups to bring about social change. PMID- 9414637 TI - Interleukin 4 responses in acute leukaemia patients with severe chemotherapy induced leucopenia. AB - T lymphocyte functions in acute leukaemia patients with severe chemotherapy induced leucopenia were investigated using 3 different approaches: (i) analysis of serum concentrations of the T cell cytokine interleukin 4 (IL4) demonstrated that serum IL4 levels increased during complicating bacterial infections. However, this response was modulated by a concomitant increase in serum levels of the potential IL4 antagonist soluble IL4 receptor alpha chain (sIL4R alpha). (ii) Even during leucopenia a subset of T lymphocytes derived from leucopenic patients expressed the activation markers CD25 (IL2 receptor), CD71 (transferrin receptor) and HLA-DR. (iii) Subsets of circulating CD4+ and CD8+ T lymphocytes could undergo clonogenic proliferation in vitro, and a majority of these clones secreted IL4. CD4+ clones showed higher IL4 levels than CD8+ clones. Our results indicate that T lymphocytes can be activated and contribute to cytokine responses in acute leukaemia patients with severe chemotherapy-induced cytopenia. PMID- 9414638 TI - Different expression of adhesion molecules on myeloid and B-lymphoid CD34+ progenitors in normal bone marrow. AB - The expression of adhesion molecules was studied on B lymphoid and myeloid CD34+ precursors in normal bone marrow. Bone marrow aspirates were labelled in a double fluorescence procedure with the CD34 monoclonal antibody 43A1 and with antibodies directed against maturation and differentiation antigens and adhesion molecules. Three clusters of CD34+ cells could be distinguished by their light scatter characteristics in flow cytometry. The population with the lowest forward scatter contained B-lymphoid precursors while the two others showed phenotypic characteristics of, respectively, early and late myeloid precursors. Nearly all CD34+ cells in the 3 subpopulations expressed VLA-4, VLA-5, LFA-3 and H-CAM. B lymphoid progenitors showed a higher density of VLA-4 and VLA-5 than the myeloid progenitors. Myeloid precursors, and particularly the late subset, expressed more HCAM than the B-lymphoid progenitors. The majority of the CD34+ cells also expressed LFA-1 and L-selectin. Higher numbers of positive cells were found in the myeloid subset. The early myeloid subset showed the highest positivity for L selectin. We conclude that B lymphoid and early and late myeloid CD34+ precursors in normal bone marrow show a different profile of adhesion molecules. These profiles could reflect a higher tendency of the myeloid CD34+ precursors to circulate. PMID- 9414639 TI - Immunoglobulin G subclasses of anti-human platelet antigen 1a in maternal sera: relation to the severity of neonatal alloimmune thrombocytopenia. AB - The monoclonal antibody immobilization of platelet antigen (MAIPA) technique was employed to detect and semiquantitatively assess total IgG anti-HPA-1a and its subclasses in sera of mothers who gave birth to severely thrombocytopenic (< 50 x 10(9) platelets/L)(n = 14) or mildly thrombocytopenic/unaffected (> 50 x 10(9) platelets/L)(n = 13) neonates. There was no statistically significant difference between the IgG anti-HPA-1a subclass composition of the 2 groups of sera. The majority of sera (26/27, 96%) showed IgG1 + IgG3 while 17/27 (63%) had all 4 subclasses of the antibody. No significant differences between the severely thrombocytopenic and mildly thrombocytopenic/unaffected groups were detected in the levels of IgG, IgG1, IgG2 or IgG4 of the antibody. However, the values of IgG3 anti-HPA-1a were significantly higher in the severely thrombocytopenic than in the mildly thrombocytopenic/unaffected group of sera with only little overlap (median 2.94 vs. 1.68, range 1.36-9.71 vs. 1.50-2.84, respectively; p < 0.01). The results suggest that maternal IgG3 anti-HPA-1a has predictive value for severe thrombocytopenia of the neonate. However, a prospective study of IgG HPA 1a subclasses in a greater number of maternal sera at different times of pregnancy is needed to test if IgG3 anti-HPA-1a is predictive of the degree of fetal/neonatal thrombocytopenia. PMID- 9414640 TI - Decrease in arterial oxygen partial pressure within the first 24 h of rhGM-CSF administration in AML patients. AB - GM-CSF may induce pulmonary complications, such as dyspnea with temporary decreases in oxygen saturation described as first dose effect for higher dosages of intravenous rhGM-CSF. This study investigated possible pulmonary disturbances in adult de novo AML patients receiving yeast rhGM-CSF 24 h prior to chemotherapy under phase II/III conditions. Eighteen patients were monitored for 22 treatment episodes. GM-CSF was given s.c. 1 q.d., 2 q.d. or continuously i.v. at 250 micrograms/m2/d 24 h prior to induction chemotherapy (TAD9, n = 18) and consolidation (TAD9, n = 4). Spirometry, bodyplethysmography, single breath diffusion capacity (DLCO) and arterial blood gas analyses were obtained prior to GM-CSF, and repeated after 24 h. Pulse oxymetric oxygen saturation (saO2) was registered continuously for the first 16 h within day 1 of rhGM-CSF treatment. Patients were aged 21-75 years. The saO2 monitoring did not reveal any first dose effect. PaO2 values decreased from 78.9 mmHg before GM-CSF to 72.8 mmHg after 24 h (p < 0.01, maximum shift 15 mmHg). PaO2 shifts occurred mainly with pre existing lowered paO2, but otherwise were independent of age, the route of GM-CSF administration, leukocyte levels, or increase of leukocytes with GM-CSF. Increases in AaDO2 reflected the paO2 shifts (p < 0.05). No dyspnea corresponded to these changes. DLCO values did not decrease significantly after 24 h. Summarily, contemporary dosage of yeast rhGM-CSF avoids short-term oxygen desaturations, but leads to clinically benign impairment in oxygen tension, based on ventilation/perfusion mismatches. This should be taken into account for patients starting at subnormal paO2. PMID- 9414642 TI - Observations on the relationship between gamma-globin chain content and globin chain precipitation in thalassaemic erythroblasts and on the composition of erythroblastic inclusions in HbE/beta-thalassaemia. AB - Electron microscope immunocytochemical studies were performed on the bone marrow from 2 patients each with beta-thalassaemia major and beta-thalassaemia intermedia and 4 patients with HbE/beta-thalassaemia, using the immunogold technique. Studies of sections that were reacted with mouse monoclonal antibodies against human gamma-globin chains showed the presence of large quantities of gamma-chains in several erythroblasts and marrow reticulocytes containing large amounts of precipitated globin chains. At least in these cells, substantial gamma chains synthesis did not protect against the formation of erythroblastic inclusions, indicating that factors other than the rate of gamma-chain synthesis influence the extent of globin-chain precipitation. Studies of sections of marrow that were reacted with a rabbit polyclonal antibody against human alpha-globin chains and a mouse monoclonal antibody against human beta-globin chains showed that the erythroblastic inclusions in HbE/beta-thalassaemia did not contain detectable quantities of HbE and consisted only of precipitated alpha-globin chains. Thus, the generally milder phenotype of HbE/beta-thalassaemia cannot be attributed to co-precipitation of HbE and excess free alpha-globin chains. PMID- 9414641 TI - Decreased erythropoietin response in Plasmodium falciparum malaria-associated anaemia. AB - One of the most serious manifestations of Plasmodium falciparum malaria is anaemia. Its established causes are increased red cell destruction and ineffective erythropoiesis. Since proinflammatory cytokines have been shown to suppress the in vitro synthesis of erythropoietin (Epo), we measured serum immunoreactive Epo in 90 Sudanese patients suffering from malaria. Even in severe cases of anaemia (blood haemoglobin < 80 g/l), serum Epo levels rarely exceeded 300 U/l. For comparison, serum Epo was increased up to 12,000 U/l in a reference group of Caucasian patients with anaemia not associated with infection. Moreover, the slope of the log Epo/haemoglobin regression line was less steep in malarial anaemia. Thus, as documented for other chronic inflammatory disorders, there is a relative lack of Epo in malaria-associated anaemia. Treatment with the antimalarial drug chloroquine may aggravate the defect in Epo production, because chloroquine inhibited Epo synthesis when tested in cell culture. PMID- 9414643 TI - No evidence for an altered mRNA expression or protein level of haematopoietic cell phosphatase in CD34+ bone marrow progenitor cells or mature peripheral blood cells in polycythaemia vera. AB - Polycythaemia vera (PV) is a myeloproliferative disorder characterized by haematopoietic progenitor cells being hypersensitive to cytokines such as erythropoietin, interleukin-3, stem cell factor and insulin-like growth factor 1, which results in an increased production of mature blood cells. The pathogenetic cellular mechanism(s) behind this hypersensitivity to cytokines is unknown, but the number of cytokine receptors and the interaction between ligand and receptor are normal in PV. Interest has therefore focused on post-receptor mechanism(s). Haematopoietic cell phosphatase (HCP) is an intracellular tyrosine phosphatase that has been demonstrated to regulate proliferative signals negatively induced by the cytokines mentioned above. Moreover, motheaten mice that genetically lack HCP have an increased amount of erythroid progenitors that are hypersensitive to Epo, and patients with familial polycythaemia have been shown to exhibit a mutation of the Epo receptor gene that includes the docking site for HCP. We therefore studied mRNA expression of HCP in pure populations of CD34+ cells, granulocytes, platelets and lymphocytes from patients with PV, chronic myeloid leukaemia (CML) or essential thrombocythemia (ET), as well as healthy controls. Using a polymerase chain reaction analysis employing specific primers for HCP, we failed to detect any abnormalities of HCP expression in PV in any of the cell populations that were examined. Moreover, HCP mRNA expression was similar in ET and CML compared to controls. Finally, Western blot analysis revealed a normal HCP protein content in PV granulocytes and platelets. We therefore conclude that neither an impaired expression of the HCP gene nor a defect in HCP protein synthesis is present in PV, and does not seem to play a role in the aetiology of this disorder. PMID- 9414644 TI - Characterization and phenotypic analysis of differentiating CD34+ human bone marrow cells in liquid culture. AB - Our current understanding of human haematopoietic stem cell biology is based in part on the characterization of human CD34+ bone marrow cell differentiation in vitro. CD34 is highly expressed on early stem cells and haematopoietic progenitor cells with clonogenic potential and is gradually lost during differentiation and commitment. However, CD71 (transferrin receptor) is expressed at low levels on early stem cells and generally increases during haematopoietic progenitor cell proliferation. We reasoned that the combination of these surface markers would provide a useful framework for the simultaneous analysis of multiple lineage differentiation of CD34+ haematopoietic progenitor cells in liquid culture. In this report, we identify the phenotype of distinct subpopulations of myeloid, erythroid and lymphoid cells in liquid suspension culture using differential expression of CD34 vs. CD71 in combination with specific lineage markers. Freshly isolated human CD34+ bone marrow cells were introduced into suspension culture and monitored over a 6-d period using 3-colour flow cytometry. This is the first demonstration that differential expression of CD34 vs. CD71 can be used to simultaneously monitor differentiation of multiple haematopoietic cell lineages in liquid suspension culture, facilitating the study of cytokine-, drug- or chemical-induced alterations in haematopoietic progenitor cell differentiation in vitro. PMID- 9414645 TI - Combined therapy with Fludarabine and cyclophosphamide in relapsed/resistant patients with B-cell chronic lymphocytic leukaemia and non-Hodgkin's lymphomas. PMID- 9414646 TI - Improved prophylaxis against overwhelming postsplenectomy infections. PMID- 9414647 TI - Richter transformation of a T cell phenotype with p53 gene mutation. PMID- 9414648 TI - Serum thymidine kinase in congenital dyserythropoietic anaemia type I and homozygous beta-thalassaemia. PMID- 9414649 TI - Concomitant diagnosis of acute myeloid leukemia (AML) and chronic lymphocytic leukemia (CLL). Importance of flow cytometry in the diagnosis of CLL without lymphocytosis accompanying AML. PMID- 9414650 TI - Helicobacter pylori extracts exhibit nicotinamide adenine dinucleotide-derived adenylation but not mono(adenosine 5'-diphosphate-ribosyl)ation of DNA ligase. AB - The issue of toxins produced by Helicobacter pylori (H. pylori) urgently requires clarification given that the bacterium causes gastric epithelial cell damage which may lead to precancerous and cancerous changes. During an investigation of the possibility of mono(adenosine 5'-diphosphate (ADP)-ribosyl)ation by H. pylori products, as observed for other bacterial toxins, we found that radioactivity of [adenylate-32P]nicotinamide adenine dinucleotide (NAD) is incorporated into an H. pylori protein of 80 kDa after incubation with crude bacterial extract. In contrast, [carbonyl-14C]NAD did not show any radioactivity incorporation. Unexpectedly, treatment of the modified protein with 0.1 N HCl, but not 0.1 N NaOH, released the AMP moiety. Such chemical properties are characteristic of bacterial DNA ligase-AMP complexes. We found that an antibody raised against Escherichia coli DNA ligase [EC 6.5.1.2] immunoprecipitated the modified 80 kDa protein. Our results indicate that incorporation of radioactivity derived from NAD into the 80 kDa protein was due to adenylation, but not mono(ADP ribosyl)ation, of the DNA ligase of H. pylori. PMID- 9414651 TI - Hepatitis C virus in pelvic lymph nodes and female reproductive organs. AB - Based on epidemiological evidence, hepatitis C virus (HCV) is thought to be involved in the etiology of not only hepatocellular carcinoma, but also lymphoproliferative diseases. In addition, our previous studies using recently developed cell culture systems that support HCV replication have indicated that HCV possesses both hepatotropism and lymphotropism. To determine whether HCV is present in extrahepatic organs, we conducted semi-quantitative reverse transcription-polymerase chain reaction for the 5' non-coding region of the HCV genome in surgical specimens (lymph nodes, ovary, uterus, peripheral blood mononuclear cells [PBMCs] and serum) from three patients with gynecological cancer. We found relatively high HCV genome titers in the lymph nodes, not in the sera, irrespective of various titers in PBMCs. These results suggest that lymph nodes may play an important role in the carrier state and the persistence of HCV infection. Moreover, contrary to expectation, high titers of the HCV genome were observed in the ovaries and the uteri, suggesting the feasibility of mother-to infant and spouse-to-spouse transmissions of HCV. PMID- 9414652 TI - Calcium and magnesium in drinking water and risk of death from colon cancer. AB - The possible association between the risk of colon cancer and the levels of calcium and magnesium in drinking water from municipal supplies was investigated in a matched case-control study in Taiwan. All eligible colon cancer deaths (1714 cases) of Taiwan residents from 1989 through 1993 were compared with deaths from other causes (1714 controls), and the levels of calcium and magnesium in drinking water of these residents were determined. Data on calcium and magnesium levels in drinking water throughout Taiwan were obtained from the Taiwan Water Supply Corporation. The control group consisted of people who died from other causes and the controls were pair-matched to the cases by sex, year-of-birth, and year-of death. The adjusted odd ratios (95% confidence interval) were 0.79 (0.64-0.98) for the group with water calcium levels between 24.4 and 42.3 mg/liter and 0.58 (0.47-0.73) for the group with calcium levels of 42.4 mg/liter or more. The adjusted odd ratios were not statistically significant for the relationship between magnesium levels in drinking water and colon cancer. The results of the present study show that there is a significant protective effect of calcium intake from drinking water against colon cancer. PMID- 9414654 TI - p16INK4 gene mutations are relatively frequent in ampullary carcinomas. AB - A high incidence of gene mutations or deletions of p16INK4, a cell cycle regulator which inhibits the activity of cyclin-dependent kinase 4/cyclin D complex and blocks the G1-to-S transition, has been reported in pancreato-biliary tract cancers. In order to investigate p16INK4 gene alterations in sporadic ampullary carcinomas, 17 sporadic ampullary carcinomas were examined. After histological diagnosis, DNA samples extracted separately from both cancerous and normal paraffin-embedded tissues were investigated. Loss of heterozygosity (LOH) was investigated utilizing 3 microsatellite markers on 9p21-22, and a mutational analysis was performed by cloning and sequencing. LOH was observed in 3 cases (17.6%) and somatic mutations with retention of heterozygosity were found in 7 cases (41.2%). Of note was that two mutations resulted in truncated incomplete proteins and one was a point mutation at the consensus site in the conserved ankyrin repeats, which would be crucial for function. Although two-hit inactivation was not evident in any of the mutation cases and further investigation would be needed to elucidate the role of altered p16INK4, these results suggest that the p16INK4 gene mutations are relatively frequent and its inactivation might be important in ampullary carcinogenesis. PMID- 9414653 TI - Induction of aberrant crypt foci and flat-type adenocarcinoma in the colons of dogs by N-ethyl-N'-nitro-nitrosoguanidine and their sequential changes. AB - Sequential endoscopic observation of dog colons was performed during colon carcinogenesis. Two beagle dogs were given suppositories containing N-ethyl-N' nitro-N-nitrosoguanidine (ENNG) every day for five months. In month 3, aberrant crypt foci (ACF), a putative preneoplastic lesion, were found in the colons of both dogs, but not in an untreated dog. The frequency of ACF increased until month 10, and then decreased. In month 9, very small lesions, less than 1 mm in diameter, which were similar to human early flat tumors, were first noticed. One of these lesions grew to about 7 mm in size without a change in its shape for 10 months. There were more than ten flat-type tumors in the two dogs, but such lesions were not found in the untreated dog. By biopsy, two of the lesions were proved to be well-differentiated adenocarcinomas histologically. Four polypoid lesions were found in one of the carcinogen-treated dogs. Thus, flat-type adenocarcinomas were induced in the dog colon by ENNG, and their development was followed by magnifying endoscopy. PMID- 9414655 TI - Overexpression of cyclin-dependent kinase-activating CDC25B phosphatase in human gastric carcinomas. AB - CDC25 phosphatases activate cyclin-dependent kinases by removing inhibitory phosphate groups on the molecules and positively regulate the cell cycle progression. The expression of CDC25A, B and C was examined in gastric carcinoma cell lines and gastric carcinoma tissues by northern blotting and immunohistochemistry. The gastric carcinoma cell lines expressed CDC25A, B and C mRNA at various levels. The expression levels of CDC25B were generally higher than those of CDC25A and C. Of the 40 gastric carcinomas, 70% of the tumors expressed CDC25B mRNA at higher levels than the corresponding normal mucosas, while 38% overexpressed CDC25A mRNA. The CDC25C expression was at very low or undetectable levels. No obvious correlation was detected between the expression of CDC25B and p53 gene mutations. Immunohistochemically, CDC25-positive tumor cells were detected in 43 (78%) of 55 gastric carcinoma cases, of which 27 (49%) were strongly positive. Strong expression of CDC25B protein was associated with advanced stage and deep invasion. Furthermore, the incidence of strong expression was significantly higher in carcinomas with nodal metastasis than in those without metastasis. These findings suggest that overexpression of CDC25B may favor development and progression and may be an indicator of malignant behavior of gastric carcinomas. PMID- 9414656 TI - Expression of interleukin-6 and its effect on the cell growth of gastric carcinoma cell lines. AB - The expression and the effect of IL-6 were examined in human gastric carcinoma cell lines to determine whether IL-6 serves as a growth stimulator. The expression of IL-6 mRNA was detected in three (TMK-1, MKN-1, MKN-7) of 8 gastric carcinoma cell lines. All three cell lines secreted IL-6 into the culture fluid, in large amounts in the cases of MKN-1 and MKN-7 cells. Scatchard plot analysis of IL-6 binding revealed that MKN-1 and MKN-7 cells had both high- and low affinity receptors. Cell growth of MKN-1 and MKN-7 cells was stimulated by IL-6, while anti-IL-6 antibody inhibited growth. The expression of IL-1 alpha mRNA by these three cell lines was induced by IL-6. IL-1 alpha increased the expression of mRNA for IL-6 by TMK-1 cells. These findings indicate that IL-6 induced by IL 1 alpha is an autocrine growth factor for some gastric carcinomas. PMID- 9414657 TI - Mapping of a breast cancer tumor suppressor gene locus to a 4-cM interval on chromosome 18q21. AB - DPC4 and DCC, putative tumor suppressor genes implicated in the genesis of several types of human cancer, lie on the long arm of human chromosome 18. We examined 200 primary breast cancers for allelic losses on chromosome 18, using 15 microsatellite markers distributed along the long arm. Allelic loss was detected most frequently (29-30%) at loci mapped to 18q21. Deletion mapping of the 34 tumors showing partial or interstitial deletions identified a commonly deleted region within the 4-cM interval flanked by D18S474 and D18S487 at 18q21.1-q21.3. Although this interval included the DPC4 and DCC genes, we excluded DPC4 from candidacy when polymerase chain reaction-single-strand conformation polymorphism analysis of each exon failed to detect abnormalities in any of the 54 breast cancers that exhibited loss of heterozygosity involving 18q. Allelic loss on 18q was found more frequently in tumors of the solid tubular histological type (24 of 55, 44%) than in other types (24 of 113, 21%) (P = 0.0049). The results suggest that a tumor suppressor gene located within the 4-cM region at 18q21, either DCC or another gene not yet identified, may play a role in the development of some sporadic breast cancers, particularly those of the solid tubular type. PMID- 9414658 TI - Significant correlation of telomerase activity in thyroid papillary carcinomas with cell differentiation, proliferation and extrathyroidal extension. AB - Telomerase activity was examined by telomeric repeat amplification protocol assay in thyroid disease states, including adenomas and carcinomas, and correlated with clinicopathological features. Of a total of 26 papillary carcinomas, 16 cases (61.5%) were positive, with the poorly differentiated subtype being predominant (P < 0.05). A significantly more shortened terminal restriction fragment length (P < 0.05), higher incidence of extrathyroidal extension (P < 0.001), and more elevated Ki-67 labeling indices (P < 0.002) were also found in telomerase positive than in telomerase-negative papillary carcinomas. Of four follicular carcinomas, 3 cases (75.0%) were positive. Positive telomerase activity in follicular adenomas (9/23 cases, 39.1%) and lymphocytic thyroiditis (12/22 cases, 54.5%) appeared to be mainly caused by infiltrating lymphocytes. However, three cases of atypical adenoma with relatively increased Ki-67 labeling indices were positive, suggesting a possibility of malignant potential. The good correlations with extrathyroidal invasiveness, Ki-67 labeling indices and poor differentiation of papillary carcinomas, established by multivariate analysis, suggest that this parameter might have potential application in the estimation of tumor progression and prognosis, and in clinical management. PMID- 9414659 TI - Increased telomerase activities in human pancreatic duct adenocarcinomas. AB - Telomerase is a key enzyme with regard to immortalization of cancer cells and increased activity has been demonstrated in various human malignant neoplasms. Since little is known of its role in pancreatic cancers, we investigated changes in telomerase activity in human pancreatic duct adenocarcinomas and compared the frequency of increased telomerase activity with the presence of K-ras gene mutations. The samples were obtained from 38 pancreatic duct adenocarcinomas and 7 tumor surrounding tissues at surgical resection. Telomerase activity was examined by telomeric repeat amplification protocol assay and terminal restriction fragment (TRF) length was examined by Southern analysis. K-ras mutation was examined by means of polymerase chain reaction-single strand conformation polymorphism analysis. Among 38 pancreatic carcinomas, 32 (84%) exhibited increased telomerase activities with no apparent relation to the histological type of tumor, tumor size, regional lymphnode involvement and distant metastasis or clinical stage. In tissue surrounding the tumor, telomerase activity was not detected. TRF length tended to be reduced in pancreatic carcinomas. Mutations of K-ras gene were found in 24 out of the 38 (63%) cases. Among the 38 cases, 14 showed increased telomerase activity without K-ras mutation and 4 cases showed K-ras mutation without telomerase activity. These results suggest that increased telomerase activity might be a sensitive genetic diagnostic marker and could be a target for future therapy of pancreatic duct carcinomas. PMID- 9414660 TI - Regression of metastatic liver tumors in rats treated with angiogenesis inhibitor TNP-470: occurrence of apoptosis and necrosis. AB - To clarify the mechanism of the reduction of metastatic liver tumors in rats treated with angiogenesis inhibitor TNP-470, the death of tumor cells was examined pathologically and ultrastructurally. Liver metastases were developed by intravenous injection of AH-130 cells. TNP-470 was given subcutaneously after tumor cell injection. Alterations in the size and number of metastatic tumors were examined at various time points, in association with the analysis of cell death pattern. The metastatic nodules were divided into 4 groups according to the morphological patterns of cell death; no cell death, scattered apoptosis, central necrosis, and diffuse necrosis. The number and size of the metastatic tumors at 2 weeks in untreated rats were larger than those in treated rats. The number of tumors in untreated rats decreased, but the tumor size increased. All rats treated with TNP-470 were alive and free from tumors after 4 weeks, whereas all the untreated rats died of liver metastases. The percentages of the tumors with necrosis in untreated rats (61.2% at 2 weeks and 100% at 4 weeks) were significantly higher than that (31.8% at 2 weeks) in treated rats (P < 0.01). The percentage of the tumors containing apoptotic cells in treated rats was significantly higher than that in untreated rats (54.5% vs. 30.6%; P < 0.05). The growth of metastatic tumors without treatment might be faster than the growth of vessels in untreated tumors, resulting in central necrosis due to ischemia. On the other hand, the reduction of metastatic liver tumors treated with TNP-470 might be caused by inhibition of angiogenesis, providing a weak ischemic stimulus which triggers apoptosis, rather than by a direct cytotoxic effect on tumor cells, because previous in vivo experiments demonstrated that TNP-470 affected endothelial cells but not tumor cells. PMID- 9414662 TI - Antitumor activity of a novel quinoline derivative, TAS-103, with inhibitory effects on topoisomerases I and II. AB - A novel quinoline derivative, TAS-103 (6-[[2-(dimethylamino)ethyl]amino]-3 hydroxy-7H-indeno[2,1-c]quinolin -7-one dihydrochloride), was developed as an anticancer agent targeting topoisomerases (topo) I and II, with marked efficacy in solid tumors. TAS-103 inhibited topo I and II (IC50: 2 microM, 6.5 microM) at a concentration similar to or lower than those of previous agents, and had a strong cytotoxic effect on P388 and KB cells (IC50: 0.0011 microM, 0.0096 microM). TAS-103 stabilized topo I and II-DNA cleavable complexes in KB cells, generating a similar amount of topo II-DNA complex to that induced by etoposide (VP-16) but a smaller amount of topo I-DNA complex than that produced by camptothecin (CPT). In the in vivo study, intermittent i.v. administration was markedly effective against s.c.-implanted murine tumors. Furthermore, TAS-103 had marked efficacy against various lung metastatic tumors, and a broad antitumor spectrum in human tumor xenografts (derived from lung, colon, stomach, breast, and pancreatic cancer). The efficacy of TAS-103 was generally greater than that of irinotecan (CPT-11), VP-16, or cis-diamminedichloroplatinum (CDDP). PMID- 9414661 TI - All-trans-retinoic acid-dependent inhibition of E-cadherin-based cell adhesion with concomitant dephosphorylation of beta-catenin in metastatic human renal carcinoma cells. AB - We previously described an in vitro invasion assay model, using a monolayer of vascular endothelial cells grown on collagen gel, that mimics the metastatic abilities of the highly metastatic human renal carcinoma cell lines, MM-1,3 and 8 and their poorly metastatic counterparts, SN12C and Cl-8. MM-1, 3 and 8 cells were observed to penetrate the monolayer of vascular endothelial cells and grew in a spreading or scattering manner with loose cell-cell contact on collagen gel or on vascular endothelial cells. SN12C and Cl-8 cells failed to penetrate and grew in a clustering manner with tight cell-cell contact. Treatment with all trans-retinoic acid (ATRA) at non-toxic concentrations induced clustering or growth of MM-1, 3 and 8 cells on collagen gel or on vascular endothelial cells with tight cell-cell contact, and inhibited penetration. The clustering induced by ATRA was virtually blocked in the presence of anti-E cadherin antibody. E Cadherin and beta-catenin were each localized mainly at the cell-cell adherent junctions of colonizing cell populations that had been treated with ATRA. While the cellular levels of E-cadherin and beta-catenin did not change significantly following ATRA treatment, the tyrosine residue of beta-catenin was rapidly dephosphorylated. The concomitant administration of Na vanadate, an inhibitor of tyrosine dephosphorylase, inhibited both the ATRA-induced clustering and the dephosphorylation of beta-catenin tyrosine. ATRA-induced clustering of MM-3 cells may be linked to the state of tyrosine phosphorylation of beta-catenin. PMID- 9414663 TI - Growth-inhibitory effects of N,N-diethyl-2-[4-(phenylmethyl)phenoxy]-ethanamine HCl combined with cisplatin on human ovarian cancer cells inoculated into nude mice. AB - In 5-day incubation of an estrogen receptor-negative human ovarian cancer cell line (KF) with N,N-diethyl-2-[4-(phenylmethyl)phenoxy]ethanamine-HCl (DPPE), the concentration of DPPE required for 50% inhibition of KF cell proliferation (IC50) was 1.7 microM. The IC50 of DPPE for inhibition of protein kinase C (PKC) activity was 3.0 microM, a similar value to those of other antiestrogens such as tamoxifen and clomiphene. DPPE also inhibited phosphorylation of mitogen activated protein kinase in KF cells. When treatment with DPPE was started 7 days after inoculation of KF cells into nude mice, 50 mg/kg DPPE alone resulted in a significant growth retardation in the early stage of tumor growth. Although 25 mg/kg DPPE showed a similar effect to 2 mg/kg cisplatin (CDDP), the combination had the most marked tumor growth-inhibitory effect. Nude mice treated with combinations of CDDP and DPPE survived significantly longer than not only untreated, but also CDDP-alone-treated mice, while 50 mg/kg but not 25 mg/kg DPPE alone had an effect comparable to that of 2 mg/kg CDDP alone. If treatment with DPPE was begun from the day after tumor inoculation, the inhibitory effect of DPPE was further enhanced, especially when combined with CDDP. If treatment with DPPE was started in nude mice with a lower tumor burden, 25 mg/kg as well as 50 mg/kg DPPE had a similar effect to 2 mg/kg CDDP, in terms of survival. When DPPE was combined with CDDP, the effect was significantly enhanced, compared to that of either alone. These treatments could be done without any adverse side effect. Thus, we conclude that DPPE has an antiestrogen action and its tumor growth inhibiting activity is enhanced on administration in combination with CDDP. PMID- 9414665 TI - MIB1-determined proliferative activity in intraductal components and prognosis of invasive ductal breast carcinoma. AB - Intraductal components of breast carcinoma may have prognostic significance. In this study, we divided 181 invasive ductal breast carcinomas into comedo and non comedo groups based on intraductal component morphology, and differences between the two groups in clinicopathological variables, including proliferative activity and survival, were assessed. Proliferative activity was evaluated by using MIB1 antibody, which reacts with the cell-proliferation-associated Ki-67 antigen, and was expressed as the number of MIB1-positive nuclei per 1000 cancer cells in intraductal components (MIB1 labeling index). We also investigated which variables had an impact on survival. The comedo group showed a significantly higher MIB1 labeling index than the non-comedo group (P < 0.0001). The differences in disease-free and overall survival between the two groups were not significant (P = 0.2477, P = 0.2069). Multivariate analysis of the entire series showed the MIB1 labeling index to be an independent prognostic factor predicting both disease-free survival and overall survival (P = 0.0160, P = 0.0035). When multivariate analysis was repeated separately for the non-comedo and comedo groups, the MIB1 labeling index remained the most important variable predicting disease-free and overall survival in the non-comedo group (P = 0.0122, P = 0.0040). Moreover, non-comedo patients with a high MIB1 labeling index had significantly shorter disease-free and overall survival than those with a low MIB1 labeling index (P = 0.0040, P = 0.0402). These findings imply that MIB1 determined proliferative activity of intraductal components is an independent predictor of survival, and is the most important predictor in non-comedo cases. PMID- 9414666 TI - Individualizing immunosuppression for heart transplantation: strategies for the next decade. PMID- 9414664 TI - p16INK4 expression is associated with the increased sensitivity of human non small cell lung cancer cells to DNA topoisomerase I inhibitors. AB - Inactivation of p16INK4, an inhibitor of cyclin-dependent kinases 4 (CDK4) and 6 (CDK6), may be essential for oncogenesis in non-small cell lung cancer (NSCLC). We examined the sensitivity of two clones of p16INK4-transfected NSCLC cell line with homozygous deletion of p16INK4, A549/p16-1 and 2, to DNA topoisomerase I (topo I) inhibitors. A549/p16-1 and -2 showed 7.7- and 9.1-fold increases in sensitivity to CPT-11 (11,7-ethyl-10-[4-(1-piperidino)-1 piperidino]carbonyloxycamptothecin ), respectively, compared with A549 cells. Ectopic p16INK4-expressing cells also showed approximately 4.0-fold increase in sensitivity to SN-38 (7-ethyl-10-hydroxycamptothecin), the active metabolite of CPT-11, compared to the parent cells. The topo I-mediated DNA relaxation activities of ectopic p16INK4-expressing cells were approximately 5 times higher than those of the parent cells. Northern and western blot analyses indicate that these increased topo I activities of ectopic p16INK4-expressing cells were due to an elevated topo I mRNA level and an increase in topo I protein. The chemosensitivity to topo I inhibitors, topo I mRNA level, protein content and activity of a p16INK4 revertant, lacking functional p16INK4, tended to be restored toward those of the parental phenotype to some extent. These results suggest that p16INK4 expression is closely associated with the increased sensitivity of ectopic p16INK4-expressing NSCLC cells to topo I inhibitors. The up-regulation of topo I mRNA level, protein content and activity may be responsible for this hypersensitivity. PMID- 9414667 TI - The immune system's role in graft loss: theoretic considerations. PMID- 9414668 TI - Antilymphocyte-antibody prophylaxis: review of the adult experience in heart transplantation. PMID- 9414669 TI - Anti-T-cell-antibody prophylaxis in children: success with a novel combination strategy of mycophenolate mofetil and antithymocyte serum. PMID- 9414670 TI - Clinical role of immunologic monitoring during OKT3 treatment. PMID- 9414671 TI - Targeting the alloimmune response for prevention of chronic graft loss: efficacy of new immunosuppressive drugs. PMID- 9414672 TI - Can histopathology guide immunosuppression for cardiac allograft rejection in the light of new techniques? PMID- 9414673 TI - Treatment of nonhemodynamic compromising rejection: conventional approaches vs individualization/new immunosuppressive drugs. AB - Mild rejection may be treated with an increase in oral CyA, but most centers give no therapy and perform a follow-up endomyocardial biopsy within 2 weeks of rejection. Focal moderate rejection is treated similarly to mild rejection; however, there is controversy as to whether focal moderate rejection exists. Asymptomatic moderate rejection is usually treated with an increase in steroids, either i.v. or oral. The therapy for asymptomatic moderate rejection is quite variable and dependent on cardiac hemodynamics, the degree of lymphocytic infiltrates, the interval after heart transplantation, and clinical findings. Symptomatic moderate rejection or severe rejection should be treated with a combination of increased steroids and cytolytic therapy. Other forms of therapy under study may provide more effective methods of treating rejection in the future. PMID- 9414674 TI - Recurrent or persistent cardiac allograft rejection: therapeutic options and recommendations. PMID- 9414675 TI - Targeting nonalloimmune-dependent pathways. AB - Medical therapy to treat and prevent this major complication has progressed slowly. The early use of diltiazem, pravastatin, or photopheresis has had reported efficacy in slowing the development of TCAD but not in preventing its development. Current ongoing multicenter studies with mycophenolate mofetil, angiopeptin, and other agents may hold promise for the future. It is clear that whatever intervention is applied, it must be started at the time of transplantation, as the cascade of events of TCAD begins at the time of surgery. Revascularization procedures, including angioplasty and CABS, have proven to be palliative and not applicable to all patients with TCAD. Currently, retransplantation remains the only viable treatment for patients with severe transplant vasculopathy. For patients undergoing retransplantation for severe TCAD, survival and the development of TCAD in the second donor heart appear acceptable compared to outcomes for patients undergoing primary heart transplantation. The ethical dilemma surrounding heart retransplantation, however, is considerable because of the scarcity of donor organs. PMID- 9414676 TI - Organ shortage: a public health crisis. What is the French state doing about it? PMID- 9414677 TI - Organ transplantations in Japan and Asian countries. PMID- 9414678 TI - The organ shortage: a public health crisis. What are Latin American governments doing about it? PMID- 9414679 TI - The Council of Europe and organ transplantation. PMID- 9414680 TI - What is the Eurotransplant Foundation doing about the organ shortage? PMID- 9414681 TI - The organ shortage: what are Australian organ sharing organizations doing about it? AB - A version of the Spanish model of organ donor generation is being trailed in Australia with early success. National implementation of this approach is underway, with the expectation that the current donor rate of 10.6 donors/pmp/year will be doubled. PMID- 9414682 TI - The organ shortage: what are organ sharing organizations doing about it? PMID- 9414683 TI - The response to the challenge of organ shortage in the Middle East region: a summary. PMID- 9414685 TI - BaltTransplant: a new organization for transplantation in the Baltic States. PMID- 9414684 TI - Views of Muslim scholars on organ donation and brain death. PMID- 9414686 TI - Organ donation relationship between US military hospitals and Japanese organ network. PMID- 9414687 TI - Legislative initiatives to increase donation. PMID- 9414689 TI - The Swedish National Donor Register. AB - The large number of individuals who have registered with the National donor register indicates that the general public feels there is a need for such a register. It also indicates that the register is accepted by the general public. Considering that the Swedish transplant act is an opting out law, it was to be expected that those objecting to donation would be overrepresented as compared to their representation in the general public. This was confirmed when the opinions of the first 300,000 persons to register were compared to a survey of attitudes made at the same time. According to the guidelines for the medical profession issued by the Board, the Register always has to be consulted in the case of a potential donor. The Register is frequently consulted and found useful by the licensed procurement coordinators. The rulings of the Swedish Data inspection board are to be followed and any divergence will be noted and acted upon by the Board. The new legislation was proposed at a time when the number of cadaver donors was declining. The number of cadaver organ donors remains unchanged. It is concluded that it is for governments to decide on donor registers and for government agencies and professionals to design safe registries and continuously supervise how they are used. The Swedish National Donor Register is safe and operational. PMID- 9414688 TI - A survey on attitudes to organ donation among three generations in a country with 10 years of presumed consent legislation. PMID- 9414691 TI - The Provincial Health Card and a computer-based organ/tissue donor registry in Canada. PMID- 9414690 TI - Legal basis for cadaver donation in Mexico: a simple diagram that facilitates the legal procedure, initial experience. PMID- 9414692 TI - The Swedish Transplant Coordinators' experience of the new Transplantation Act and the donor register 1 year after implementation. AB - The register has been a support for the coordinators in Sweden when the relatives have said one thing and the deceased person another in regards to donation. Most of the staff are positive to the register and the safety regulations around it. Most often it has been a relief for the relatives that the deceased person's wishes also have been documented in the register or in the donor card. Most decisions are still, however, made by relatives. There is still a need for more positive information about transplantation and its value. PMID- 9414693 TI - Development of a national initiative to increase the number of kidneys for transplant purposes in Canada. PMID- 9414694 TI - Why does Mecklenburg-Vorpommern have the highest number of organs available for transplantation within the Eurotransplant Community? PMID- 9414695 TI - A success story: promoting the incorporation of donation- and transplantation related content in nursing school curricula. PMID- 9414696 TI - Survey of concepts and attitudes among healthcare professionals towards organ donation and transplantation. PMID- 9414697 TI - Improving attitude and knowledge of healthcare professionals towards organ donation in Israel: results of 12 European donor hospital education programs. PMID- 9414699 TI - Unified strategy for public education in organ and tissue donation. PMID- 9414698 TI - Organ donation: the influence of personal attitude on professional behavior. PMID- 9414700 TI - Walking the walk: behavior shifts to match attitude toward organ donation- Richmond, Virginia, 1994-1996. PMID- 9414701 TI - Public education postpresentation research. PMID- 9414702 TI - MORE life. PMID- 9414703 TI - The role of hospital-based family support teams in improving the quality of the organ donation process. PMID- 9414704 TI - Financial and legislative issues: selected topics. PMID- 9414705 TI - Financial and legislative issues. PMID- 9414706 TI - Clinical and economic outcomes of expanded criteria donors in renal transplantation. AB - Although graft and patient survival rates were similar between recipients of kidneys from ECDs and non-ECDs, transplantation with organs from ECDs was significantly more expensive. Multivariate analysis using stepwise linear regression demonstrated length of stay to be a strong proxy for total hospital costs. Inherent tensions between the overall good clinical outcomes associated with the use of ECDs in terms of graft survival and the markedly increased costs seen with these organs are evident. PMID- 9414707 TI - Estimating the economic impact of an initiative to increase organ donation in a transplant center. PMID- 9414708 TI - The Texas Nondonor Hospital Project: a preliminary report on the impact of inhouse coordinators on organ donation rates in nondonor hospitals. PMID- 9414709 TI - Public feelings about financial incentives for donation and concern about incurring expenses due to donation in one US city. PMID- 9414710 TI - Solidarity model as nonmonetary incentive could increase organ donation and justice in organ allocation at the same time. PMID- 9414711 TI - Results of single kidneys from donors aged 9 to 60 months: results in 144 adult recipients. PMID- 9414712 TI - Selection strategies for successful utilization of less than 15.kg pediatric donor kidneys. PMID- 9414713 TI - Should pediatric donors younger than 2 years of age be used in kidney transplantation? PMID- 9414714 TI - Do the factors of the renal graft donor have an effect on early graft function and long-term graft survival? PMID- 9414715 TI - Experience with transplantation of elderly donor kidneys. PMID- 9414716 TI - Long-term function of single pediatric kidneys less than 48 months of age transplanted into adult recipients compared with adult cadaveric and living related transplants. PMID- 9414718 TI - Outcome of patients undergoing orthotopic liver transplantation with elderly donors (over 60 years). PMID- 9414717 TI - Extending the boundaries of acceptable organ donors: a means of expanding the donor pool for liver transplantation. PMID- 9414719 TI - Donor age, organ import, and cold ischemia: effect on early outcomes after simultaneous kidney-pancreas transplantation. PMID- 9414720 TI - Evaluating the donor pool: impact of using hearts from donors over the age of 49 years. PMID- 9414721 TI - Increasing the donor pool: recovery of hearts from older donors. PMID- 9414722 TI - The donor supply. PMID- 9414723 TI - Trends in organ donation. AB - Presumed consent legislation produces more donors and in particular more organs per donor. In cadaveric donation, the number of elderly donors is increasing, so the quality of organs available is getting poorer. There will be no substantial rise in transplantation unless there is a major breakthrough in the current programs. There will be an increase in the proportion of living (un)related donors and debate about ethics of transplantation. Adverse publicity and debate may affect cadaveric donation and refusal rates. PMID- 9414725 TI - Developing a benchmark of organ donation rates for assessing hospital performance. PMID- 9414724 TI - Trends in organ donation, recovery and disposition: UNOS data for 1988-1996. PMID- 9414726 TI - New Donation Program at a Mexican social security institution: a Mexican model of cadaver donation and organ sharing--initial experience. PMID- 9414727 TI - Pancreas underutilization in the United States: analysis of United Network for Organ Sharing data. PMID- 9414728 TI - Underutilization of the potential for cadaver pancreas donation. PMID- 9414729 TI - Eight grafts in seven patients from a single cadaver donor: a case report. PMID- 9414730 TI - Lack of disease transmission using donors with fatal encephalitis. PMID- 9414732 TI - Procurement of transplantable organs from brain dead transplant recipients. PMID- 9414731 TI - Single kidney transplants from donors with low estimated creatinine clearance. PMID- 9414733 TI - Renal transplantation using hepatitis B or C serology-positive donors. PMID- 9414734 TI - Organ procurement in Rio Grande do Sul: analysis of two periods of activity. PMID- 9414735 TI - Xenotransplantation: a potential solution to the shortage of donor organs. PMID- 9414736 TI - Clinical impact of infections in left ventricular assist device recipients: the importance of site and organism. PMID- 9414737 TI - A novel paracorporeal mechanical assist device for newborns and infants allows bridging to transplantation. PMID- 9414738 TI - Influence of HLA matching and DNA typing on kidney and heart transplant survival in black recipients. PMID- 9414739 TI - Non-heart-beating cadaver organ procurement: two remaining issues. PMID- 9414740 TI - Organ procurement after evidence of brain death in victims of acute poisoning. PMID- 9414741 TI - A liver transplant candidate (fulminant hepatic failure from amanita phalloides poisoning) as a multiorgan donor. PMID- 9414742 TI - Multimodality evoked potentials as a valuable technique for brain death diagnosis in poisoned patients. PMID- 9414743 TI - Ischemic damage prevention by N-acetylcysteine treatment of the donor before orthotopic liver transplantation. PMID- 9414744 TI - Improvement in early function of the hepatic graft after treatment of the donor with N-acetylcysteine: clinical study. PMID- 9414745 TI - Impact of viral infections on organ donation in South Africa. PMID- 9414746 TI - Influence of thyroid function in brain stem death donors on kidney allograft function. PMID- 9414747 TI - Effect of nutritional procurement for the donor liver on Kupffer cell activation in porcine liver transplantation. PMID- 9414748 TI - Modulation of Kupffer cells affects the metabolism of protease inhibitor administered in cold-preserved liver transplantation. PMID- 9414749 TI - Long-term fasting of donors deteriorates mitochondrial adenosine triphosphate synthesis in liver grafts during cold preservation. PMID- 9414750 TI - Intraoperative inhaled nitric oxide during anesthesia for lung transplant. PMID- 9414751 TI - Hemodynamics during inhaled nitric oxide in lung transplant candidates. PMID- 9414752 TI - The effect of donor gender on renal allograft survival and influence of donor age on posttransplant graft outcome and patient survival. PMID- 9414754 TI - Preliminary experience with double kidney transplants from adult cadaveric donors: analysis of United Network for Organ Sharing data. PMID- 9414753 TI - Impact of donor gender on graft survival after liver transplantation. PMID- 9414755 TI - Transplantation of elderly donor kidneys into young adults. PMID- 9414756 TI - Long-term outcome of kidney transplantation from expanded criteria donors: a single center experience. PMID- 9414758 TI - Donor characteristics and hospital survival in cardiac transplantation. The Spanish Groups of Heart Transplantation. PMID- 9414757 TI - Impact of organ preservation techniques on outcome of cardiac transplantation in the era of donor pool expansion. PMID- 9414759 TI - Myocardial protection during heart transplantation using blood cardioplegia. PMID- 9414760 TI - Orthotopic heart transplantation with bicaval anastomosis using older donors. PMID- 9414761 TI - Transplant recipients as organ donors: the domino transplant. PMID- 9414762 TI - Different patterns of cadaver heart utilization in two similar organ procurement organizations. PMID- 9414763 TI - The donor organ shortage in the Balkans: accept everyone! PMID- 9414765 TI - Laparoscopic live donor nephrectomy removes disincentives to live donation. PMID- 9414764 TI - Analysis of 160 consecutive living unrelated kidney transplants: 1983-1997. PMID- 9414766 TI - Kidneys from living donors with renal vascular disease may be safely used for transplantation. PMID- 9414767 TI - Donor-transmitted IgA nephropathy: long-term follow-up of kidney donors and recipients. PMID- 9414768 TI - Comparison of cadaveric and living liver donors. PMID- 9414769 TI - Donor selection for living-related liver transplantation. PMID- 9414770 TI - Assessment of pretransplantation warm ischemia time by phosphorus-31 magnetic resonance spectroscopy in pig kidneys. PMID- 9414771 TI - Long-term cold ischemia reduces nitric oxide metabolism in reperfused rabbit kidneys. PMID- 9414772 TI - Rewarming gradients in porcine kidney grafts during simulated second warm ischemic time. PMID- 9414773 TI - Warm ex vivo perfusion prevents reperfusion injury in warm ischemically damaged kidneys. PMID- 9414774 TI - Predictors of graft outcome in warm ischemically damaged organs. PMID- 9414775 TI - Posttransplant management following warm ischemic injury. PMID- 9414776 TI - Autotransplantation of the kidney in primates: a model of renal damage to study the ischemia-reperfusion injury. PMID- 9414777 TI - A new form of donor allocation: Mid-German Transplant Union. PMID- 9414778 TI - Impact of donor and recipient gender on liver transplantation. PMID- 9414779 TI - HLA-DRB1 genotyping in prospective selection of kidney donors. PMID- 9414780 TI - Impact of changes in the UNOS renal allocation system. PMID- 9414781 TI - Expanded mandatory sharing of zero-antigen mismatched kidneys: first-year effects following policy revision. PMID- 9414782 TI - Fate of Canadian organs offered to the United States. PMID- 9414783 TI - Characteristics of kidneys offered as paybacks in the zero-antigen mismatch sharing policy. PMID- 9414784 TI - The impact of improvements in procurement and utilization on US liver recipients. PMID- 9414785 TI - Effect of liver preservation on hepatocyte calcium and ATP regeneration. PMID- 9414786 TI - Alpha glutathione S-transferase as novel parameter for hepatocellular damage in the isolated perfused rat liver. PMID- 9414787 TI - Machine perfusion of the liver: maintenance of mitochondrial function after 48 hour preservation. PMID- 9414788 TI - Mechanisms of preservation and reperfusion injury in human liver transplantation. PMID- 9414789 TI - Influence of warm ischemia time on initial graft function in human liver transplantation. PMID- 9414790 TI - Randomized study on in situ liver perfusion techniques: gravity perfusion vs high pressure perfusion. PMID- 9414791 TI - Elimination of Kupffer cells and administration of protease inhibitor improve graft viability and prevent reperfusion injury in NHBD. PMID- 9414792 TI - Optimizing early graft function by a combined approach of donor and recipient conditioning in a model of rat liver transplantation. PMID- 9414793 TI - Evaluation of the superiority of subzero nonfreezing storage using isolated rat hepatocytes. PMID- 9414794 TI - Efficacy of endothelin-1 receptor antagonist for protecting the function of grafted livers from preservation-reperfusion injury in pigs. PMID- 9414795 TI - Blood flow and oxygen extraction during normothermic recirculation and total body cooling predict viability of liver from non-heart-beating pig donors. PMID- 9414796 TI - Histological changes during and after liver transplantation from non-heart beating donor pig. PMID- 9414797 TI - Orthotopic liver transplantation from non-heart-beating cadaver donors in the rat: a preliminary study. PMID- 9414798 TI - Short-term restoration by gaseous oxygen for long-term preservation of predamaged livers from non-heart-beating donors. PMID- 9414799 TI - Usefulness of a combination of machine perfusion and pentoxifylline for porcine liver transplantation from non-heart-beating donors with prolonged hypotension. PMID- 9414800 TI - Liver transplantation with organs from non-heart-beating donors. PMID- 9414801 TI - Adenine nucleotide liver tissue concentrations from non-heart-beating donor pigs and organ viability after liver transplantation. PMID- 9414802 TI - Evaluation of ischemic liver injury during graft procurement from non-heart beating donor pigs. PMID- 9414804 TI - Changes in adenine nucleotides and lipid hydroperoxides during normothermic cardiopulmonary bypass in a porcine model of type II non-heart-beating donor. PMID- 9414803 TI - Value of MEGX test in predicting survival after liver transplantation from non heart-beating donor pigs. PMID- 9414805 TI - Kidney transplantation from asystolic cadaveric donors. PMID- 9414806 TI - The effect of machine perfusion preservation on early function of non-heart beating donor kidneys. PMID- 9414807 TI - Factors influencing short- and long-term survival of kidneys transplanted from non-heart-beating donors. PMID- 9414808 TI - The non-heart-beating donor--survival factors. PMID- 9414809 TI - Is xenogeneic cytotoxicity of plasma from patients with hepatic failure to porcine hepatocytes less than that in healthy human volunteers? PMID- 9414810 TI - The new immunosuppressive malononitrilamide MNA 279 prolongs skin xenograft survival in a mouse-to-rat model. PMID- 9414811 TI - Long-term xenograft survival by combination therapy of malononitrilamide MNA 715 with cyclosporine. PMID- 9414812 TI - Analysis of xenogeneic microchimerism in hamster-to-rat lung xenotransplantation. PMID- 9414813 TI - Sperm-mediated gene transfer: production of pigs transgenic for a human regulator of complement activation. PMID- 9414814 TI - Improving early graft function: role of preservation. PMID- 9414815 TI - Triple strategy to improve early graft function after heart transplantation. PMID- 9414816 TI - Hepatocyte apoptosis and cytosolic calcium dynamics in ischemic injury. PMID- 9414818 TI - Ex vivo resuscitation of kidneys following postmortem warm ischemia. PMID- 9414817 TI - Evaluation of liver function and morphology following ischemia-reperfusion injury in pigs. PMID- 9414819 TI - Experimental and clinical assessment of preservation-induced reperfusion injury comparing renal transplant blood flow and renal endothelin concentrations. PMID- 9414820 TI - Pancreas preservation with HTK solution in the pig. PMID- 9414821 TI - Mitochondrial ischemia-reoxygenation injury and plasma membrane integrity in human endothelial cells. PMID- 9414822 TI - A role for the mitochondrial permeability transition pore in ex vivo hypothermic resuscitation of warm ischemic kidney tissue. PMID- 9414823 TI - Evaluation of a portable hypothermic microperfusion system for storage of the donor heart: clinical experience. PMID- 9414824 TI - Evaluation of unstored hearts: immediate functional differences between St Thomas, UW, and cardiosol following cardioplegia. PMID- 9414825 TI - 24-hour preservation of the newborn myocardium: a comparison of two solutions. PMID- 9414826 TI - Evaluation of mitochondrial function after cold preservation of pancreatic islet cells from donors treated with pefloxacin. PMID- 9414827 TI - Kidney preservation in pigs using celsior, a new organ preservation solution. PMID- 9414828 TI - Improved liver preservation: high-K+ UW versus high-Na+ UW. PMID- 9414830 TI - Transplantation of kidneys from non-heart-beating donors: retrospective analysis of the outcome. PMID- 9414829 TI - What solutions are best? Overview of flush solutions. PMID- 9414831 TI - Outcome of referrals to a non-heart-beating kidney retrieval team over a 5-year period. PMID- 9414832 TI - Utilization of kidneys from non-heart-beating donors by portable cardiopulmonary bypass. PMID- 9414833 TI - Type I non-heart-beating donors: policy and results. PMID- 9414834 TI - A rapid organ recovery program for non-heart-beating donors. AB - We report a successful method for rapid organ recovery from the non-heart-beating donor, which can open a new resource of organs for transplantation. The RORP is not controversial, is simple in design and execution, and results in kidneys that are viable for transplantation. Special personnel and equipment are needed but are easily incorporated in the overall budget of an OPO or donor hospital. Clearly more research is needed to rebuild ischemically damaged cells ex vivo and to develop new agents/methods to minimize the reperfusion response. When these processes are better understood and managed, the full potential of the NHBD as a donor resource will be fully achieved. We agree with others that the donor shortage could be entirely relieved by routine organ recovery from NHBD trauma victims. PMID- 9414835 TI - Comparison of techniques for rapid cooling of organs in a non-heart-beating porcine model. PMID- 9414836 TI - An overview of current non-heart-beating donor transplantation. PMID- 9414837 TI - Clinical outcome of cadaveric renal transplantation with non-heart-beating donors: special reference to serious complications. PMID- 9414838 TI - Comparative study of the use of systolic and asystolic kidney donors between 1981 1995 in La Coruna, Spain. PMID- 9414839 TI - Are kidneys from non-heart-beating donors second class organs? PMID- 9414840 TI - Evaluation of the factors related to early graft function in 90 kidney transplants from non-heart-beating donors. PMID- 9414841 TI - Quadruple immunosuppressive therapy with low-dose cyclosporine provides superior kidney transplant outcome using grafts of non-heart-beating uncontrolled cadavers. PMID- 9414842 TI - Marginal kidney graft function from non-heart-beating donors. PMID- 9414843 TI - Pulsatile preservation in renal transplantation. PMID- 9414844 TI - Reduced renal vascular injury following warm ischemia and preservation by hypothermic machine perfusion. PMID- 9414845 TI - An assessment of ischemic injury of the kidney for transplantation during machine pulsatile preservation. PMID- 9414846 TI - Pulsatile preservation characteristics predict early graft function in extended criteria donor kidneys. PMID- 9414848 TI - Usefulness of continuous hypothermic perfusion preservation for renal grafts from non-heart-beating donors with prolonged warm ischemia time. PMID- 9414847 TI - Beneficial effect of aspirin and protease inhibitor as graft conditioning using machine perfusion in warm ischemically injured kidneys. PMID- 9414849 TI - A fully automated organ perfusion workstation: the LifeSustainer 1000. PMID- 9414851 TI - Does the quality of early graft function influence the long-term outcome of renal transplantation? PMID- 9414850 TI - Release of alpha-glutathione S-transferase (alpha GST) and pi-glutathione S transferase (pi GST) from ischemic damaged kidneys into the machine perfusate- relevance to viability assessment. PMID- 9414852 TI - The role of cold ischemia on graft survival in recipients of renal transplants. PMID- 9414853 TI - Effect of waiting time and dialysis on outcomes in simultaneous kidney-pancreas transplant recipients. PMID- 9414854 TI - Role of donor/recipient Na+/H+ antiport activity as a nonimmunologic predictor of kidney graft outcome. PMID- 9414855 TI - Survival on hemodialysis versus renal transplantation following primary renal allograft failure. PMID- 9414856 TI - Pretreatment of kidney allografts with anti-CD5 immunotoxin: a new chance for high responder recipients. PMID- 9414857 TI - Results of liver transplantation in function of certain risk factors associated with the indication: an approach to quality control? PMID- 9414858 TI - Liver transplantation by preservation of the caval flow with temporary porto caval shunt or veno-venous bypass. PMID- 9414859 TI - Lymphocyte sub-population content of the liver graft before liver transplantation. PMID- 9414860 TI - Organ shortage: viability of potential organ donors and possible loss depend on health care workers who are responsible for the organ procurement program. PMID- 9414861 TI - Immunosuppression in auxiliary partial liver transplantation with FK506 in rats. PMID- 9414862 TI - Expanding the donor pool: the challenge of non-heart-beating donor kidneys. PMID- 9414863 TI - Incidence of tumors in organ transplants. PMID- 9414864 TI - The role of hepato-coeliac arterial reconstruction in porcine segmental pancreatic autotransplantation. PMID- 9414865 TI - Bioethics and organ transplantation. PMID- 9414866 TI - Living unrelated (paid) renal transplantation: besides the ethics. PMID- 9414867 TI - An experimental model of acute cerebral death: Tc-99m bicisate brain imaging and kinetics. PMID- 9414868 TI - Anencephalic neonates and diagnosis of death. PMID- 9414869 TI - Effect of organ exenteration procurement on pancreas function. PMID- 9414870 TI - Outcome of renal transplantation with kidneys from marginal donors (preliminary communication). PMID- 9414871 TI - A comparison study of the Belzer machine preservation solution with and without penicillin. PMID- 9414872 TI - Comparison of static versus pulsatile preservation of matched-paired kidneys. PMID- 9414873 TI - Intensive care nurses' participation in organ procurement: impact on organ donation rates. PMID- 9414874 TI - Living renal donation in the African-American community. PMID- 9414875 TI - RNA transcription and lysosomal activation during xenotransplant hyperacute rejection: inhibition of RNA polymerase. AB - In summary, our data indicates that AFB1 has no effect on LyAct during XTHAR. Although these results are from short-term incubation studies, they nevertheless suggest that inhibition of RNA transcription by AFB1 may be of no consequence in LyAct during XTHAR. Hence, our results appear to support the view that, in general, XTHAR recruits preformed components and may not require de novo synthesis of lysosomal proteins. Additional studies using long-term incubation (days to weeks) and other protein synthesis blockers are suggested to further elucidate the effects of protein synthesis inhibition of LyAct during XTHAR. PMID- 9414876 TI - Extracellular calcium concentration influences lysosomal behavior in the in vitro xenotransplant hyperacute rejection model. PMID- 9414877 TI - Kidney transplantation utilizing donors from both age extremes. PMID- 9414879 TI - Liver transplantation in African-Americans with chronic alcoholic liver disease: focus on the elevated cardiac index pretransplantation. PMID- 9414878 TI - Lysosomal activation in xenotransplantation: an early, calcium-sensitive process. PMID- 9414880 TI - Accepting prolonged ischemia times for the donor heart. PMID- 9414882 TI - Issues of informed consent and access to extended donor pool kidneys. PMID- 9414881 TI - Extended donor criteria: current donors with a history of prolonged insulin dependent diabetes mellitus. PMID- 9414883 TI - Decreased rejection episodes in African-American renal transplant recipients receiving mycophenolate mofetil/tacrolimus therapy. PMID- 9414884 TI - Expanded criteria for donor kidneys: an update on outcome in single versus dual kidney transplants. PMID- 9414886 TI - The Swedish National Donor Register. AB - The large number of individuals who have registered with the National donor register indicates that the general public feels there is a need for such a register. It also indicates that the register is accepted by the general public. Considering that the Swedish transplant act is an opting out law it was to be expected that those objecting to donation would be overrepresented as compared to their representation in the general public. This was confirmed when the opinions of the first 300,000 to register was compared to a survey of attitudes made at the same time. According to the guidelines for the medical profession issued by the Board, the Register always has to be consulted in the case of a potential donor. The Register is frequently consulted and found useful by the licensed procurement coordinators. The rulings of the Swedish Data inspection board are to be followed and any divergence will be noted and acted upon by the Board. The new legislation was proposed at a time when the number of cadaver donors was declining. The number of cadaver organ donors remains unchanged. It is concluded that it is for governments to decide on donor registers and for government agencies and professionals to design safe registries and to continuously supervise how they are used. The Swedish National Donor Register is safe and operational. PMID- 9414885 TI - Utilization of cadaveric kidneys from donors with reactive hepatitis C virus antibody or reactive hepatitis B core antibody. PMID- 9414887 TI - Successful multiorgan retrieval following prolonged donor resuscitation and primary myocardial infarction diagnosis. PMID- 9414888 TI - Hemodynamic heterogeneity of multiorgan donors in Poland. PMID- 9414889 TI - Surgical approaches for expanded organ usage in liver transplantation. PMID- 9414890 TI - The Fourth international Samuel L. Kountz Symposium on renal disease and transplantation in African Americans: foreword. PMID- 9414891 TI - The Samuel L. Kountz Award. John Ruffin. PMID- 9414892 TI - OKT3 induction therapy and cadaveric renal transplantation in black patients. PMID- 9414893 TI - HLA antigens, alleles, and haplotypes among African-Americans. PMID- 9414894 TI - Genetic polymorphisms of HLA class I and class II system in the Basque population. PMID- 9414895 TI - Genes within and flanking the major histocompatibility region are risk factors for diabetes, insulin resistance, hypertension, and microalbuminuria in African American women. PMID- 9414897 TI - African-Americans with type I insulin-dependent diabetes mellitus and end-stage renal disease: results after simultaneous pancreas-kidney transplantation. PMID- 9414896 TI - Increased inducible interleukin-2 receptor expression on CD8 lymphocytes in black recipients of renal allografts with deteriorating function. PMID- 9414898 TI - Simultaneous pancreas-kidney transplantation in Hispanic recipients with type I diabetes mellitus and end-stage renal disease. PMID- 9414899 TI - Hypertension does not contribute to end-stage renal disease in black recipients of kidney/pancreas transplants. PMID- 9414900 TI - Relative risk of policystic kidney disease in HLA-DR1 basque individuals. PMID- 9414901 TI - Results of renal transplantation in black patients in South Africa. PMID- 9414902 TI - The living donor process in kidney transplantation: influence of race and comorbidity. PMID- 9414903 TI - Impact of a large nonindigenous population on the renal transplant waiting list. PMID- 9414904 TI - Renal transplantation in African-American recipients: three decades at a single center. PMID- 9414905 TI - Race is not a critical factor in orthotopic liver transplantation. PMID- 9414906 TI - Outcome of kidney transplantation in African-Americans using tacrolimus. PMID- 9414907 TI - Influence of HLA and CREG matching in African-American primary cadaver kidney recipients: UNOS 1991-1995. PMID- 9414908 TI - Relationship between hope and self-esteem in renal transplant recipients. PMID- 9414909 TI - Postrenal transplant health beliefs and ethnicity. The Compliance Study Group. PMID- 9414910 TI - Ethnic and gender variations in postrenal transplant physical symptoms. PMID- 9414911 TI - Organ donation among Hispanics: a single-center experience. PMID- 9414912 TI - Barriers to donation in minority, low-income, and rural populations. PMID- 9414914 TI - Attitudes of Korean-Americans in and around New York City toward organ transplantation. PMID- 9414913 TI - Increasing living kidney donation in African Americans. PMID- 9414916 TI - Community education and empowerment key to increased minority donation rates. PMID- 9414915 TI - A success story in minority donation: the LifeGift/Ben Taub General Hospital In House Coordinator Program. PMID- 9414917 TI - Racial impact: increasing minority consent rate by altering the racial mix of an organ procurement organization. PMID- 9414918 TI - Progress in availability of donors of color: the National Marrow Donor Program. PMID- 9414919 TI - Alternatives to organ transplantation. PMID- 9414920 TI - Detrimental effects of brain death on the potential organ donor. PMID- 9414921 TI - Donor management: state of the art. PMID- 9414922 TI - Technical aspects of carotid endarterectomy with Hemashield patch graft. AB - In this review I describe my method of carotid endarterectomy with synthetic patch angioplasty. The use of Hemashield patch angioplasty is relatively new in my practice, and since I have adopted it I feel it has reduced or eliminated early restenosis and acute postoperative occlusion. The technique is simple and straightforward, and does not require any special preparation of the patch material. The patch is stronger and safer than saphenous vein. The other steps of the endarterectomy are unchanged, and the same suture can be used for patch placement. I also describe here a new commercial shunt which I have developed and which I feel is an improvement over the shunts I previously used. I place an indwelling shunt selectively, if electroencephalography criteria indicate that a shunt is needed. In my series this occurs in 15% of cases, increasing to 25% in patients with contralateral carotid artery occlusion. The overall stroke rate in my carotid artery series is 1.8% at present. PMID- 9414923 TI - Hemispheric cerebral atrophy after traumatic extra-axial hematoma in adults. AB - The relationship between traumatic extra-axial hematomas and cerebral atrophy was investigated in 42 adult patients aged between 15 and 50 years who required removal of extra-axial hematomas. These patients were followed up by serial computed tomography for more than 6 months after head injury. Nine of these patients developed cerebral atrophy. Their Glasgow Coma Scale score on admission was 6.4 +/- 2.8 (mean +/- SD). The score of the patients without cerebral atrophy was 9.6 +/- 3.3 (p < 0.01). These patients had three extradural and six subdural hematomas. All patients with cerebral atrophy had cerebral swelling postoperatively, more prominent in the hemisphere ipsilateral to the hematoma in seven patients. This swelling was associated with global hypodensity and persisted for 10.4 +/- 2.9 days. The severity of cerebral atrophy was more prominent in the hemisphere ipsilateral to the hematoma in five of these seven patients. Extra-axial hematoma in patients with severe head injury can induce hemispheric cerebral atrophy in the underlying cerebral hemisphere. PMID- 9414924 TI - Effect of glycerol on blood flow distribution in tumoral and peritumoral brain tissue. AB - The effect of glycerol on blood flow in tumoral and peritumoral tissue was measured in 32 patients with brain tumor, 17 gliomas and 15 meningiomas. Blood flow before and after the administration of glycerol was measured by stable xenon enhanced computed tomography. The tumor part of glioma was significantly hypoperfused. In contrast, the tumor part of meningioma was significantly hyperperfused. Peritumoral edema of both glioma and meningioma was hypoperfused. After the administration of glycerol, blood flow increased in all regions except for the tumor part of glioma. Vascular responses to glycerol may be different in these two tumor types. The steal phenomena of blood flow might occur in cases of glioma. PMID- 9414925 TI - Staged transsphenoidal surgery for fibrous nonfunctioning pituitary adenomas with suprasellar extension. AB - Staged transsphenoidal surgery was performed in seven patients with nonfunctioning pituitary adenomas with suprasellar extension. Remnant adenomas were present in a supersellar position after complete removal of the intrasellar tumor, and did not descend into the sella because of the fibrous nature in five patients or fibrous nature and dumbbell shape in two. Magnetic resonance images were obtained every 2 weeks following initial surgery. The suprasellar residual adenomas descended into the sella within 2 months in six patients and 1.5 months in one patient. A second transsphenoidal operation was performed 2 months following the initial procedure in four patients, 3 months in one, and 5 months in two. In six of the seven patients, extensive tumor removal was achieved safely and easily by the staged approach. Patients were followed up over 6 to 58 months (mean +/- SD 24.7 +/- 18.9 months). There were no major surgical complications or recurrence of tumor on follow-up images. Our postoperative imaging studies and surgical results demonstrated that staged transsphenoidal surgery is an effective and safe treatment for fibrous nonfunctioning pituitary adenomas with suprasellar extension. PMID- 9414926 TI - Three-dimensional computed tomography imaging of a frontal skull base fracture. AB - A 7-year-old boy presented with recurrent meningitis after head trauma. Coronal computed tomography (CT) revealed prolapse of the intracranial soft tissue into the right ethmoidal sinus, leading to the diagnosis of right frontal skull base fracture. Three-dimensional CT with bone windows provided a lifelike image of the fracture lateral to the right olfactory groove. This image was most useful in the preoperative planning of the repair surgery. PMID- 9414927 TI - Fenestration of the basilar artery associated with persistent primitive trigeminal artery. AB - A 39-year-old male presented with a putaminal hemorrhage. Angiography disclosed the incidental findings of association of basilar artery fenestration and persistent primitive trigeminal artery. This is the first report of this association of congenital anomalies of the cerebral vessels. PMID- 9414928 TI - Carotid endarterectomy for radiation-induced carotid artery stenosis. AB - A 60-year-old male presented with radiation-induced left carotid artery stenosis. Carotid endarterectomy was performed successfully without postoperative deficits. Carotid endarterectomy is the therapeutic management of choice for these lesions. PMID- 9414930 TI - Successful removal of meningioma of the pineal region after embolization. AB - A 69-year-old female presented with a meningioma of the pineal region manifesting as gait disturbance and mental dysfunction. Magnetic resonance imaging revealed a homogeneously well-enhanced circumscribed round mass of about 5 cm in diameter in the pineal region. Angiography demonstrated that the tumor was fed mainly by the bilateral middle meningeal arteries (MMAs), and preoperative intravascular embolization was performed through the bilateral MMAs using estrogen-alcohol and polyvinyl acetate. The tumor was very soft and easily totally resected via the right occipital transtentorial approach. Preoperative embolization is a very useful technique to facilitate removal of deep-seated tumors. PMID- 9414929 TI - Hypothalamic pilocytic astrocytoma presenting with intratumoral and subarachnoid hemorrhage. AB - A 45-year-old male presented with sudden onset of severe headache. Computed tomography and magnetic resonance imaging demonstrated an irregularly enhanced suprasellar mass with intratumoral and subarachnoid hemorrhage. The mass was removed in two operations. Histological examination of the tumor revealed pilocytic astrocytoma. The relatively rich vascularity and perivascular tumor cell proliferation observed in this benign lesion were probably the causes of this extremely rare association. PMID- 9414931 TI - Intraventricular, cystic, atypical meningioma. AB - A 37-year-old male presented with a very rare cystic meningioma in the trigone of the left lateral ventricle. Neurological examination revealed mild conduction aphasia and right hemisensory disturbance. Computed tomography and magnetic resonance imaging showed a solid, enhancing tumor in the left trigone which had multiple cystic components located anteriorly and superiorly. The tumor was totally resected via a parietal, transventricular approach. Histological examination revealed an atypical meningioma with cellular pleomorphism and prominent nucleoli. Both the solid component and the cyst wall consisted of tumor cells. PMID- 9414933 TI - Experimental biology and NASA in the twenty-first century. PMID- 9414932 TI - Intracranial epidermoid cyst including elements of old hematoma. AB - A 70-year-old female presented with symptoms of right-sided trigeminal neuralgia. Computed tomography showed a high-density mass in the prepontine cistern without enhancement. Magnetic resonance (MR) imaging showed the mass as heterogeneous with variable but largely high-signal intensity on T1-weighted images and low signal intensity on T2-weighted images. At surgery, the lesion was found to be an epidermoid cyst filled with old blood and lipid debris. The high-signal intensity on the T1-weighted images may reflect lipid or methemoglobin with the low intensity on T2-weighted images representing hemosiderin. Most intracranial epidermoid cysts appear as low-intensity lesions on T1-weighted images and high signal intensity on T2-weighted images. Typical MR imaging findings are neither specific for nor constant with epidermoid cysts, requiring critical differential diagnosis. PMID- 9414934 TI - Is male infertility on the increase? PMID- 9414935 TI - Conflicts in resuscitation: ethical dilemmas. Paper presented at a joint meeting of Ulster Medical Society and Ulster Neuropsychiatric Society 20 February 1997. PMID- 9414937 TI - Visual impairment in Northern Ireland. AB - Statistics on the registration of blind and partially-sighted patients in Northern Ireland underestimate the true extent of visual impairment within our community. In comparison to other UK regions, where between 0.53% and 0.59% of the population avail of blind or partial sight registration, only 0.35% of residents in Northern Ireland appear on the respective registers. Most patients on the combined registers are in the older age groups and many also suffer from other disabilities. Regional discrepancies may be attributed to a combination of factors including: patient attitudes to the registration process, medical attitudes to registration and local anomalies in the way in which social services departments both record and present annual registration returns. Better liaison is necessary between the community, hospital and voluntary sector providers to improve identification and support services for the visually impaired in the future. PMID- 9414936 TI - Prescription of oxygen concentrators and survival in Northern Ireland. AB - Long-term oxygen therapy (LTOT) has been shown to prolong survival and to improve quality of life in patients with chronic obstructive pulmonary disease (COPD) and in respiratory failure. In Northern Ireland oxygen concentrators have been available on prescription since August 1986, initially on a restricted basis from hospital physicians only. This was followed by open prescribing from April 1989, when concentrators could be prescribed by general practitioners. This study examined prescribing habits of LTOT during both periods, and patient survival. Case notes of all prescriptions of oxygen concentrators in Northern Ireland (to April 1991) were reviewed. Prescription criteria and advice regarding usage during both periods were analysed. A questionnaire survey of subjects during open prescribing documented the advice given at the time of prescription and current usage. 164 charts of 178 total installations were available for review. During both periods many concentrators were installed without adherence to the prescribing criteria at the time (75% restricted, 48% open). The majority of these were on the advice of a consultant respiratory physician and only 14 were prescribed directly by GPs. 89 of 91 subjects receiving current LTOT during the study period completed questionnaires. Of the subjects prescribed LTOT during the restricted period, 2 subjects are still alive (median survival 19 m, range 0 104). From the open period, survival data was available on 107 of 129 subjects with 17 still alive (median survival 22 m, range 0-94). This study documents an inadequate rate of prescribing and a lack of conformity to guidelines in the provision of LTOT in Northern Ireland. We would suggest that familiarisation with the prescribing criteria, formal written advice at the time of prescription, appropriate follow up to ensure adequate supplementation and regular patient education on the use of LTOT would address these problems to a substantial degree. PMID- 9414938 TI - The Unlinked Anonymous HIV Prevalence Monitoring Programme in N. Ireland 1992 1995. AB - Previous evidence has suggested that Northern Ireland is a low seroprevalence area for HIV infection. The Unlinked Anonymous HIV Prevalence Monitoring Programme initiated in England and Wales in 1990 was extended to Northern Ireland in 1992. Patients attending the Genitourinary Medicine Clinic at the Royal Victoria Hospital have, with informed consent, been tested anonymously for HIV infection since that time. The results of the survey between 1992 and 1995 have shown an overall seroprevalence rate 3.01% for homosexual/bisexual men, 0.08% for heterosexual men, and 0.05% for heterosexual women. These results confirm the previous impression of low HIV seroprevalence in Northern Ireland and the survey provides an excellent longitudinal study by which changes may be monitored. PMID- 9414939 TI - A survey of exercise based cardiac rehabilitation services in Northern Ireland. AB - A survey was undertaken to establish the extent of provision of phase III exercise-based cardiac rehabilitation in Northern Ireland. Detailed information was obtained on patient referral mechanisms, patient assessment, the exercise component of cardiac rehabilitation and the use of outcome measures. The results suggest that cardiac rehabilitation in Northern Ireland has developed on an ad hoc basis, and although most centres accept myocardial infarction and coronary artery bypass graft patients for cardiac rehabilitation, higher risk patients are generally excluded from these programmes. Currently, little in the way of standard outcome measures are being used to evaluate the effectiveness of existing cardiac rehabilitation services. This paper makes several recommendations to facilitate the development of a more standardised service within Northern Ireland. PMID- 9414940 TI - Transabdominal cervicoisthmic cerclage: initial experience. AB - Pregnancy outcome before and after insertion of transabdominal cervical cerclage is evaluated. The series also reports on the first cases of second pregnancies with the original suture left in situ. It is our view that transabdominal cervical cerclage should only be performed in units that have specialists in Perinatal Medicine. PMID- 9414942 TI - Living in interesting times. PMID- 9414941 TI - Fine needle aspiration cytology of gastric carcinoma. AB - Four patients between 58 and 81 years of age undergoing investigation and endoscopic biopsy for gastric carcinoma also were subjected to direct-vision fine needle aspiration cytology of their mucosal lesions which yielded malignant cells. The relevance of this technique is discussed regarding both intrinsic and extrinsic lesions of the gastrointestinal tract. PMID- 9414943 TI - Dispensaries in counties Armagh and Down in the pre-Famine years. AB - This paper traces the development of dispensaries in the counties of Armagh and Down in the decades prior to the Great Famine. It examines the number and distribution of dispensaries and discusses their management, finance and daily administration. The role of dispensary doctors, their conditions of employment and the diseases which they treated are also considered. PMID- 9414944 TI - Thyroidectomy for medullary carcinoma in MEN 2A: positive genetic screening as the sole indicator for surgery. PMID- 9414946 TI - Solitary breast metastasis from carcinoma of colon. PMID- 9414945 TI - Cutaneous metastasis as a complication of hepatic intra-arterial chemotherapy. PMID- 9414947 TI - Thoracic duct cyst in supraclavicular region. AB - A 28-year-old female attended an outpatient clinic in October, 1989, because of a tumor in the left supraclavicular fossa, detected in a health examination. Following exploratory puncture of the tumor which yielded milky-white fluid, suggesting a cyst in the thoracic duct, she was admitted to our department. The cyst was unilocular measuring about 6 cm in diameter, and the fluid content was chyle-rich in lipids. Lymphography demonstrated a lymphatic structure adjacent to the lesion and scattered lymph vessels on the cyst surface. On November 16 the cyst was resected. A restiform structure was observed between the cyst and the thoracic duct, but the presence or absence of communication was unclear. The histological diagnosis was thoracic duct cyst. Thoracic duct cyst occurring in the cervical region is very rare. Our case may provide useful information as to its pathogenesis and the mode of retention of cyst fluid. PMID- 9414948 TI - Fluoxetine induced bradycardia in presenile dementia. PMID- 9414949 TI - Papillomaviruses: progress for human and veterinary medicine. PMID- 9414950 TI - Equine neurodegenerative diseases--stressed neurons and other radical ideas. PMID- 9414951 TI - Bovine papillomavirus and cancer. AB - Bovine papillomavirus (BPV) induces papillomas of cutaneous or mucosal epithelia in cattle. The papillomas are benign tumours and generally regress, but occasionally persist and provide the focus for malignant transformation to squamous cell carcinoma, particularly in the presence of environmental cofactors. This has been experimentally demonstrated for BPV-2 and cancer of the urinary bladder, and BPV-4 and cancer of the upper alimentary canal in cattle feeding on bracken fern. In this review, several aspects of the biology of the virus are described including viral genome structure, regulation of transcription of the viral oncogenes, function of the viral oncoproteins, cooperation between virus and chemical cofactors in carcinogenesis, virus latency and prophylactic and therapeutic vaccination programmes. PMID- 9414952 TI - Aerosol therapy in the equine species. AB - Inhalation therapy plays an increasing role in the management of equine respiratory disorders. This alternative to systematic treatment permits a high concentration of medication to act locally while minimizing side effects and residues. In human medicine, literature in this field is prolific and continuously renewed, whereas in veterinary medicine, applications of aerosol therapy are less extensive. This review considers the principles of action of the different types of devices used for inhalation, i.e., nebulization, metered-dose inhalation and dry powder inhalation, describes the technical and practical requirements for their use in the equine species and considers the advantages and disadvantages of each inhalation device. The pharmacological agents currently administered to horses by inhalation are also discussed. Perspectives of aerosol therapy in the equine species, including aerosols already used in human medicine and their potential applications for horses are described. PMID- 9414953 TI - Association between plasma vitamin E concentration and the risk of equine motor neuron disease. AB - Equine motor neuron disease (EMND) is a neurodegenerative disorder of the somatic lower motor neurons that results in a syndrome of diffuse neuromuscular disease in the adult horse. The aetiology of this disorder is unknown, although prior studies have suggested that a deficiency in the lipid antioxidant vitamin E (alpha-tocopherol) contributes to the development of EMND. This paper describes a case-control study designed to investigate the association between plasma vitamin E levels and the risk of EMND for horses. Signalment, plasma vitamin E levels at the time of referral, and information relative to dietary and management practices were collected from 53 horses diagnosed with EMND and 69 controls. The mean plasma vitamin E concentration in EMND cases was significantly lower than that of control horses. After controlling for other risk factors of EMND, there was a statistically significant association between plasma vitamin E levels and EMND, with the likelihood of the disease increasing as the vitamin E concentration decreased. These findings support the reported role of vitamin E deficiency as one of the risk factors for EMND. PMID- 9414954 TI - Claw lesions in dairy cattle: development of sole and white line haemorrhages during the first lactation. AB - The development of claw haemorrhages was monitored in first-calving dairy heifers from 4 weeks before calving to 32 weeks after calving. Before calving, lesions were few but the number, extent and severity of claw haemorrhages increased following simultaneous calving and housing. Lesions were most severe in the white line at 9 weeks, and in the sole at 14 weeks, but recovery began while the animals were still housed. That the increase in white line lesions after calving, and the subsequent recovery preceded that of the sole, suggests that the pathogenesis of the lesions may differ in these two anatomical regions. It is proposed that an initial insult to the corium primarily affects the laminar region and that corium damage increases with the resulting alteration in the physical forces on the sole. PMID- 9414957 TI - Biological weapons: an increasing threat. AB - The background to the risk of biological warfare is examined with particular reference to recent developments in biotechnology and genetic engineering. The provisions of and problems with the Biological Weapons Convention are discussed, with particular reference to verification. The role of doctors and scientists in issues related to research on biological warfare are considered; they should do more to inform the general public and press for appropriate action. PMID- 9414958 TI - Acoustic anti-personnel weapons: an inhumane future? AB - High intensity sound is foremost among newer technologies being considered by the military for use in war weaponry and in control of civil disorder, particularly in the United States where some weapons may be nearing deployment. The various forms of acoustic energy and the weapons in which they could be applied are considered, and the adverse effects on humans described; these can be serious and persistent, and are potentially lethal. Military applications for which high energy sound has been contemplated in the past or are being considered for the future are examined, and some existing research projects reviewed. Such uses of acoustic energy in war are criticized in the light of the obligation imposed by existing international humanitarian law for weapons not to cause 'unnecessary suffering' or 'superfluous injury'. The humanitarian community must advocate greater humanitarian protection in warfare. PMID- 9414955 TI - Cervico-thoracic vertebral osteomyelitis in 14 calves. AB - Fourteen cases of cervico-thoracic (C-T) vertebral osteomyelitis in calves were investigated over a 6 year period. The onset of clinical signs was between 2 and 9 weeks of age. There was no breed or sex predisposition. The clinical history prior to referral extended from 5 days to 8 weeks (mean 20 days). The most common clinical presentation was difficulty in rising with a tendency to knuckle or kneel on the forelimbs which displayed hypotonia and hyporeflexia. In over half the cases pain could be elicited on manipulation of the neck. The lesion in all cases involved one or more of the vertebrae from C6 to T1. The diagnosis was confirmed by radiology and/or at post mortem. Four animals were discharged after treatment, 10 animals were humanely destroyed. Salmonella dublin was isolated from the vertebral lesion in eight of the 10 calves at post mortem. PMID- 9414956 TI - Comparison of carprofen and flunixin meglumine as adjunctive therapy in bovine respiratory disease. AB - In an open, controlled, multi-centre clinical field trial, seven 'naturally occurring' outbreaks of acute febrile (rectal temperature > or = 39.5 degrees C) respiratory disease in housed calves were treated with a single antimicrobial agent, and either the non-steroidal anti-inflammatory drug (NSAID) carprofen (n = 95) or flunixin meglumine (n = 92) on an alternate basis. Carprofen was administered as a single subcutaneous injection at a mean dosage of 1.4 mg kg-1 (range 1.2 to 1.9 mg kg-1) body weight on the first day and flunixin meglumine by intravenous injection at a mean dosage of 2.0 mg kg-1 (range 1.2 to 2.6 mg kg-1) body weight on the first 3 consecutive days. All calves were examined clinically immediately prior to initial treatment and on three occasions up to 1 week after the end of treatment. There were no statically significant differences between NSAID groups in reduction of clinical parameters between examinations, or in overall efficacy. This trial demonstrated that a single dose of carprofen was equally effective as three daily doses of flunixin meglumine as adjunctive therapy to antimicrobial treatment in acute respiratory disease in calves. PMID- 9414959 TI - The psychological, social and economic consequences of blinding soldiers. AB - Blindness producing multiple disabilities in a cognitively intact patient is qualitatively different from other war injuries. It merits individual consideration in international humanitarian law. PMID- 9414960 TI - The importance of the E-cadherin-catenin complex in the maintenance of intestinal epithelial homoeostasis: more than intercellular glue? PMID- 9414961 TI - Intestinal metaplasia at the squamocolumnar junction in patients attending for diagnostic gastroscopy. AB - BACKGROUND: The incidence of adenocarcinoma of the oesophagus and gastric cardia is increasing rapidly. Barrett's oesophagus is the major risk factor. Intestinal metaplasia at the squamocolumnar junction in the absence of Barrett's oesophagus is common but its relation to adenocarcinoma and gastro-oesophageal reflux disease is unclear. AIMS: To study the prevalence and clinical, endoscopic, and histological associations of intestinal metaplasia at the squamocolumnar junction. METHODS: Biopsy specimens were taken from 120 randomly selected patients undergoing routine diagnostic endoscopy. Eight biopsy specimens, taken from above and below the squamocolumnar junction, gastric fundus, and gastric antrum, were stained with haematoxylin/eosin, alcian blue/periodic acid-Schiff, and Gimenez, and graded independently by one pathologist. RESULTS: Intestinal metaplasia at the squamocolumnar junction was found in 21 patients (18%). Metaplasia was associated with increasing age (p < 0.01) and antral intestinal metaplasia (p = 0.04). Logistic regression analysis revealed that age was the only independent predictor (p < 0.01). There was no association with symptomatic, endoscopic, or histological markers of gastro-oesophageal reflux disease. CONCLUSIONS: Intestinal metaplasia at the squamocolumnar junction is a common finding. It is associated with increasing age but not gastro-oesophageal reflux disease. PMID- 9414962 TI - Stimulus and site specific induction of hiccups in the oesophagus of normal subjects. AB - BACKGROUND: Hiccups that are induced by a large meal have been suggested to result from gastric overdistension. The role of the oesophagus in precipitating hiccups has never been defined. AIMS: To determine the involvement of oesophageal mechanoreceptors in the hiccup reflex. METHODS: Ten normal healthy subjects were prospectively evaluated at a university affiliated hospital. Controlled inflation of a polyethylene bag in the proximal and distal oesophagus was carried out using slow ramp and rapid phasic distensions, by an electronic distension device. RESULTS: Hiccups were induced in four subjects only during rapid phasic distensions and only in the proximal oesophagus. The mean (SEM) minimal volume threshold for the hiccup reflex was 32.5 (4.8) ml, which was above the perception threshold. Hiccups appeared during inflation and resolved after deflation. CONCLUSIONS: Sudden rapid stretch of the mechanoreceptors in the proximal oesophagus can trigger the hiccup reflex in normal subjects. Only rapid distensions above a determined volume threshold will predictably induce hiccups in a given subject. This mechanism may play a role in the physiological induction of hiccups. PMID- 9414964 TI - Oesophageal motor responses to gastro-oesophageal reflux in healthy controls and reflux patients. AB - AIMS: To compare oesophageal motor responses to gastro-oesophageal reflux (GOR) in 16 healthy controls (group 1) and 25 reflux patients, 15 without (group 2) and 10 with (group 3) oesophagitis. METHODS: All subjects underwent 24 hour ambulatory oesophageal pH measurements (5 cm above the lower oesophageal sphincter (LOS)) combined with pressure monitoring (5, 10, and 15 cm above the LOS for oesophageal body motility and 27 cm above the LOS for voluntary swallow detection). Contraction patterns (peristaltic, simultaneous, isolated, mixed type, and non-transmitted swallows) and peristaltic contraction wave characteristics (amplitude, duration, and velocity) during GOR were compared in the three groups. RESULTS: The average number of motor activities per minute was significantly higher in group 1 (p < 0.05). In all groups, the most common motor contraction pattern was peristaltic. The percentage of peristaltic activity per subject was significantly higher in group 1 (p < 0.05). There were no significant differences in other contraction patterns among the three groups (p > 0.05). Of the peristaltic contraction wave characteristics there were no significant differences in any parameters (amplitude, duration, and velocity) among the three groups (p > 0.05). The average pH increment in response to motor activities was significantly higher in group 1 (p < 0.05). CONCLUSIONS: Motor responses to GOR were found to be predominantly primary peristaltic in all groups. During GOR, reflux patients have less frequent activity, a smaller proportion of activity is peristaltic, and the average pH increment in response to motor activities is reduced compared with controls. PMID- 9414963 TI - Associations between different forms of gastro-oesophageal reflux disease. AB - AIMS: To study the epidemiology and natural history of gastro-oesophageal reflux disease (GORD). METHODS: Retrospective cohort study involving all 172 hospitals of the Department of Veterans Affairs. A total of 194,527 patients with GORD were followed between 1981 and 1994. Distribution of oesophagitis, oesophageal ulcer, oesophageal stricture, strictured hiatus hernia, hiatus hernia, and pyrosis by age, sex, and ethnicity were determined. The comorbid occurrence of various forms of GORD in identical patients was analysed by an age and race standardised morbidity ratio. The population of all hospitalised veterans was used for comparison. RESULTS: Severe forms of GORD associated with oesophageal erosions, ulcers, or strictures, affected elderly, white, male patients more often than their corresponding opposite demographic group. All forms of GORD clustered in the same patient population; on average, any form of GORD was 10 times more likely to occur in a patient with another form of GORD than without. The highest morbidity ratio (22) was found in oesophageal ulcer and stricture. About one third of all patients with oesophageal erosions, ulcers, or strictures also had hiatus hernia; 46% of patients with hiatus hernia were diagnosed as having other forms of GORD. While one third of all oesophageal strictures appeared in patients without other forms of GORD diagnosed at any time, oesophageal ulcers were always associated with some other form of GORD. No clear cut progression in different forms of GORD was found. CONCLUSIONS: Older age, male sex, and white ethnicity are risk factors in the development of severe forms of GORD. The most severe grade of GORD is reached at the onset of the disease. PMID- 9414965 TI - Influencing the practice and outcome in acute upper gastrointestinal haemorrhage. Steering Committee of the National Audit of Acute Upper Gastrointestinal Haemorrhage. AB - AIMS: To assess changes in practice and outcome in acute upper gastrointestinal haemorrhage following the feedback of data, the reemphasis of national guidelines, and specific recommendations following an initial survey. DESIGN: A prospective, multicentre, audit cycle. Forty five hospitals from three health regions participated in two phases of the audit cycle. PATIENTS: Phase I: 2332 patients with acute upper gastrointestinal haemorrhage; phase II: 1625 patients with upper gastrointestinal haemorrhage. METHODS: Patients were evaluated with respect to management (with reference to the recommendations in the national guidelines), mortality, and length of hospital stay. RESULTS: Following the distribution of data from the first phase of the National Audit and the formulation of specific recommendations for improving practice, the proportion of hospitals with local guidelines or protocols for the management of upper gastrointestinal haemorrhage rose from 71% (32/45) to 91% (41/45); 12 of the 32 hospitals with guidelines during the first phase revised their guidelines following the initial survey. There was a small but significant increase in the proportion of all patients who underwent endoscopy (from 81% to 86%), the proportion who underwent endoscopy within 24 hours of admission (from 50% to 56%), and the use of central venous pressure monitoring in patients with organ failure requiring blood transfusion or those with profound shock (from 30% to 43%). There was, however, no change in the use of high dependency beds or joint medical/surgical management in high risk cases. There was no significant change in crude or risk standardised mortality (13.4% in the first phase and 14.4% in the second phase). CONCLUSIONS: Although many of the participating hospitals have made efforts to improve practice by producing or updating guidelines or protocols, there has been only a small demonstrable change in some areas of practice during the National Audit. The failure to detect any improvement in mortality may reflect this lack of change of practice, but may also reflect the fact that a large proportion of the deaths in this unselected study are not preventable; only a very large study could hope to demonstrate a significant change out of the context of a clinical trial. PMID- 9414966 TI - Specific adaptation of gastric emptying to diets with differing protein content in the rat: is endogenous cholecystokinin implicated? AB - BACKGROUND: Recent studies indicate that gastric emptying may be influenced by patterns of previous nutrient intake. Endogenous cholecystokinin (CCK), whose synthesis and release can be affected by dietary intake, has a major role in the regulation of gastric emptying. AIMS: To evaluate the influence of diets with differing protein content on gastric emptying of differing liquid test meals and plasma CCK levels in the rat and to check whether the inhibitory effect of exogenous CCK on gastric emptying is modified after long term intake of diets with differing protein content. METHODS: Rats were fed for three weeks with high protein, medium protein (regular), or low protein diet. On day 22 gastric emptying of a peptone meal was studied. In addition, basal and postprandial CCK levels after the different dietary regimens were measured by bioassay. The time course of dietary adaptation was studied and its specificity assessed through the use of different (peptone, glucose, and methylcellulose) test meals. The effect of exogenous CCK-8 on gastric emptying was studied at the end of the adaptation period (three weeks). RESULTS: Feeding the animals with a high protein diet for three weeks resulted in a significant (p < 0.05) acceleration (by 21.2 (8.2)%) of gastric emptying while feeding with a low protein diet was followed by a significant (p < 0.05) delay (by 24.0 (6.2)%) in the emptying rate. When the time course of the effect of dietary adaptation on gastric emptying was studied, it appeared that at least two weeks are required for dietary protein to be effective. The regulatory effect of dietary protein on gastric emptying proved to be dependent on meal composition. Only the emptying rate of a protein containing meal (40% peptone) was significantly modified by previous dietary intake. No significant (p > 0.05) changes were observed with glucose and methylcellulose meals whose emptying rates were similar in rats receiving a high protein or low protein diet. A peptone meal strongly and significantly (p < 0.05) increased plasma CCK levels in rats fed a medium protein (regular) diet. Results were similar in rats receiving a low protein diet (p < 0.05) but not in rats on a high protein diet (p > 0.05). As a consequence, postprandial plasma levels of CCK in rats fed with a medium or low protein diet were significantly (p < 0.05) higher than those in rats receiving a high protein diet. In rats on high and low protein diets, dose response curves to CCK-8 were virtually identical, suggesting that dietary protein intake has no influence on the effect of exogenous CCK. CONCLUSIONS: These results clearly show that gastric emptying of a protein containing meal can be modified by previous dietary protein intake. This effect, which is time dependent and meal specific, may be related to changes in endogenous CCK release which will affect emptying rate. While the exact mechanisms underlying this adaptive response need to be studied and clarified further, these results emphasise the importance of dietary history in the evaluation and interpretation of gastric emptying data. PMID- 9414967 TI - Antigastric autoantibodies in Helicobacter pylori infection: implications of histological and clinical parameters of gastritis. AB - BACKGROUND: It has recently been shown that humoral antigastric autoreactivities occur in a substantial number of Helicobacter pylori infected patients. AIMS: To analyse the relevance of such antigastric autoantibodies for histological and serological parameters of the infection as well as for the clinical course. METHODS: Gastric biopsy samples and sera from 126 patients with upper abdominal complaints were investigated for evidence of H pylori infection using histology and serology. Autoantibodies against epitopes in human gastric mucosa were detected by immunohistochemical techniques. Histological and clinical findings of all patients were then correlated with the detection of antigastric autoantibodies. RESULTS: H pylori infection was significantly associated with antigastric autoantibodies reactive with the luminal membrane of the foveolar epithelium and with canalicular structures within parietal cells. The presence of the latter autoantibodies was significantly correlated with the severity of body gastritis, gastric mucosa atrophy, elevated fasting gastrin concentrations, and a decreased ratio of serum pepsinogen I:II. Furthermore the presence of anticanalicular autoantibodies was associated with a greater than twofold reduced prevalence for duodenal ulcer. CONCLUSION: The data indicate that antigastric autoantibodies play a role in the pathogenesis and outcome of H pylori gastritis, in particular in the development of gastric mucosal atrophy. PMID- 9414968 TI - Abnormal postprandial duodenal chyme transport in patients with long standing insulin dependent diabetes mellitus. AB - BACKGROUND: Patients with long standing diabetes mellitus frequently have upper gut dysmotility. Gastroparesis has been well studied, whereas detailed data on duodenal motor function are limited. AIMS: To characterise postprandial duodenal chyme transport in such patients. METHODS: Intraluminal multiple impedance measurement, recently introduced as a novel technique for investigation of chyme transport, was used to study postprandial duodenal chyme flow in 10 patients with long standing insulin dependent diabetes mellitus with gastroparesis, and 10 healthy volunteers. RESULTS: Four distinct transport patterns of chyme, termed bolus transport events (BTEs), were found in both groups and could be characterised as: short distance propulsive; simple long distance propulsive; retrograde; and complex long distance propulsive. Diabetic patients had significantly lower numbers of propulsive BTEs (p < 0.01), and higher proportions of retrograde BTEs and complex long distance BTEs (p < 0.05) than control subjects, whereas the proportion of simple long distance BTEs was significantly lower (p < 0.05). The mean propagation velocities of the BTEs were similar in both groups. CONCLUSION: Abnormal postprandial duodenal chyme transport was found in patients with long standing insulin dependent diabetes mellitus. This is characterised by transport disorganisation and may result in disturbed chyme clearance. PMID- 9414969 TI - Improved clinical tolerance to chronic lactose ingestion in subjects with lactose intolerance: a placebo effect? AB - BACKGROUND: Uncontrolled studies of lactose intolerant subjects have shown that symptom severity decreases after chronic lactose consumption. Adaptation of the colonic flora might explain this improvement. AIMS: To compare the effects of regular administration of either lactose or sucrose on clinical tolerance and bacterial adaptation to lactose. METHODS: Forty six lactose intolerant subjects underwent two 50 g lactose challenges on days 1 and 15. Between these days they were given 34 g of lactose or sucrose per day, in a double blind protocol. Stool samples were obtained on days 0 and 14, to measure faecal beta-galactosidase and pH. Symptoms, breath H2 excretion, faecal weight and electrolytes, and orofaecal transit time were assessed. RESULTS: Except for faecal weight, symptoms were significantly milder during the second challenge in both groups, and covariance analysis showed no statistical difference between them. In the lactose group, but not in the sucrose group, faecal beta-galactosidase activity increased, pH dropped, and breath H2 excretion decreased. CONCLUSION: Bacterial adaptation occurred when lactose intolerant subjects ingested lactose for 13 days, and all symptoms except diarrhoea regressed. Clinical improvement was also observed in the control group which displayed no signs of metabolic adaptation. This suggests that improved clinical tolerance may be just a placebo effect. PMID- 9414970 TI - Enhanced mucosal permeability and nitric oxide synthase activity in jejunum of mast cell deficient mice. AB - BACKGROUND: Recent reports have described a modulating influence of nitric oxide (NO) on intestinal mucosal permeability and have implicated a role for mast cells in this NO mediated process. AIMS: To assess further the contribution of mast cells to the mucosal permeability changes elicited by the NO synthase (NOS) inhibitor NG-nitro-L-arginine methylester (L-NAME), using mast cell deficient (W/Wv) and mast cell replete mice (+/+). METHODS: Chromium-51 EDTA clearance (from blood to jejunal lumen), jejunal NOS and myeloperoxidase (MPO) activities, and plasma nitrate/nitrite levels were monitored. RESULTS: The increased EDTA clearance elicited by intraluminal L-NAME in W/Wv mice (4.4-fold) was significantly greater than the response observed in control (+/+) mice (1.8 fold). The exacerbated response in W/Wv mice was greatly attenuated by pretreatment with either dexamethasone (1.3-fold) or the selective inducible NOS inhibitor, aminoguanidine (1.4-fold), and partially attenuated by the mast cell stabiliser, lodoxamide (2.9-fold). Jejunal inducible NOS activity was significantly higher in W/Wv than in +/+ mice, while jejunal MPO was lower in W/Wv mice than in +/+ mice, suggesting that the higher inducible NOS in W/Wv does not result from the recruitment of inflammatory cells into the gut. The higher inducible NOS activity in the jejunum of W/Wv was significantly reduced by dexamethasone treatment. CONCLUSIONS: Our results suggest that mast cells normally serve to inhibit inducible NOS activity tonically in the gut and that inhibitors of NOS elicit a larger permeability response when this tonic inhibitory influence is released by mast cell depletion. PMID- 9414971 TI - Antisecretory factor suppresses intestinal inflammation and hypersecretion. AB - BACKGROUND: Antisecretory factor (AF) is a recently identified regulatory protein which inhibits the intestinal fluid secretion induced by cholera toxin. AIMS: To test the effect of AF on: (a) inflammation and hypersecretion induced by toxin A from Clostridium difficile; and (b) morphological changes and hypersecretion induced by okadaic acid (the blue mussel toxin) in rat intestinal mucosa. METHODS: Morphological changes and fluid accumulation were observed in intestinal loops challenged with 1 microgram of toxin A or 3 micrograms of okadaic acid administered before or after injection of 0.1 microgram of recombinant AF (rAF). RESULTS: The cytotoxic and inflammatory reaction caused by toxin A was abolished after treatment with rAF given either intraveneously or intraluminally prior to the toxin or one hour after the toxin. The intestinal fluid response induced by toxin A and okadaic acid was reduced 55-80% by rAF. However, the characteristic increase in goblet cells at the tips of villi in the okadaic acid treated mucosa was not inhibited by rAF. CONCLUSION: Results suggest that AF might be involved in protection against inflammation and in counteracting dehydration caused by enterotoxins. Both effects are probably mediated via the enteric nervous system. PMID- 9414972 TI - Search for Mycobacterium paratuberculosis DNA in orofacial granulomatosis and oral Crohn's disease tissue by polymerase chain reaction. AB - BACKGROUND: Although intestinal Crohn's disease has long been suspected to have a mycobacterial cause, possible mycobacterial involvement in orofacial granulomatosis (OFG) and oral lesions of Crohn's disease has not yet been investigated. AIMS: As the slow growing Mycobacterium paratuberculosis has been implicated in the aetiology of intestinal Crohn's disease, the potential involvement of this mycobacterial species in OFG and oral lesions of Crohn's disease was investigated. PATIENTS: To attempt detection of the organism in OFG and oral Crohn's disease tissue samples, a polymerase chain reaction (PCR) assay was used on archival formalin fixed, paraffin wax embedded oral tissue sections from 30 patients with OFG, seven with Crohn's disease, and 12 normal controls. METHODS: The PCR assay used was based on primers targeting the 5' region of the multicopy IS900 DNA insertion element of the M paratuberculosis genome. In order to achieve maximum sensitivity, two rounds of PCR were carried out and amplicons confirmed by Southern blot hybridisation to a digoxigenin labelled IS900 DNA probe. RESULTS: None of the OFG and oral lesions of Crohn's disease samples were positive for M paratuberculosis and all normal controls were also negative. CONCLUSIONS: These results suggest that M paratuberculosis may not be a major aetiological agent in OFG or oral Crohn's disease lesions, although the use of paraffin wax embedded tissue as opposed to fresh tissue as a sample source could underestimate the true prevalence of the organism. PMID- 9414973 TI - Imbalance of the interleukin 1 system in colonic mucosa--association with intestinal inflammation and interleukin 1 receptor antagonist [corrected] genotype 2. AB - BACKGROUND: Interleukin 1 (IL-1) alpha and beta are potent cytokines which play key roles in inflammation. They are controlled by IL-1 receptor antagonist (IL 1ra). AIMS: To investigate the influence of mucosal inflammation and IL-1ra genotype on the IL-1ra:IL-1 balance. PATIENTS AND METHODS: IL-1 alpha, IL-1 beta, and IL-1ra were measured by enzyme linked immunosorbent assay (ELISA) in biopsy specimens taken from inflamed and non-inflamed colon of 60 patients with Crohn's disease (CD), 34 with ulcerative colitis (UC), 15 inflammatory controls, and 103 non-inflamed controls. IL-1ra genotype was determined by polymerase chain reaction and gel electrophoresis. RESULTS: IL-1 alpha and IL-1 beta were significantly increased in inflamed mucosa in inflammatory bowel disease (IBD) (CD: 53.5 (22.4) and 409.9 (118.7) pg/mg protein, respectively; UC: 18.9 (6.8) and 214.5 (78.2) pg/mg, respectively) and non-IBD patients (19.2 (7.4) and 281.4 (121.0) pg/mg, respectively; p < 0.0001) compared with normal controls (2.8 (0.6) and 30.6 (5.6) pg/mg, respectively). In CD IL-1 alpha and beta were also significantly increased in non-inflamed mucosa (6.1 (1.3) pg/mg and 88.7 (17.4) pg/mg, respectively; p < 0.0012). IL-1ra:(IL-1 alpha+beta) ratios were significantly decreased in inflamed mucosa of patients with CD (182 (45); p < 0.0001), UC (425 (136); p = 0.0018) and without IBD (221 (76); p < 0.0001), and in non-inflamed mucosa in CD (369 (149); p < 0.0001) compared with normal controls (1307 (245); p < 0.0001). Patients with IL-1ra genotype 2 had slightly but significantly reduced mucosal IL-1ra concentrations (p = 0.003). The greatest difference was seen in colonic biopsy specimens from patients with inflamed Crohn's disease. CONCLUSION: Mucosal inflammation can modulate the balance of the IL-1:IL-1ra system in colonic mucosa. PMID- 9414974 TI - The distal colon provides reserve storage capacity during colonic fluid overload. AB - BACKGROUND: In addition to its absorptive function the capacity of the colon to retain fluid might be relevant in compensating for increased fluid loads and prevention of diarrhoea. The distal colon is considered to be mainly a conduit without extensive storage function. AIMS: To evaluate colonic volume capacity in a model of pure osmotic diarrhoea. METHODS: A non-absorbable, iso-osmotic solution (OS) containing polyethylene glycol (500 ml) was infused into the caecum of nine healthy volunteers; the control group (n = 5) received an equal amount of an easily absorbable electrolyte solution (ES). Fluids were radiolabelled with technetium-99m and gamma camera images were obtained for 48 hours. Counts in the proximal and distal colon were measured and regional and overall colonic transit and stool output were quantified. RESULTS: After OS, in contrast to ES, faecal output was increased significantly (p < 0.05), but colonic transit after OS was not different from transit after ES (p > 0.05). This indicates storage of OS in the colon: after OS infusion, counts in the proximal colon decreased linearly while the distal colon stored approximately 30% of radioactivity for the whole 48 hour study period. After OS, stool output correlated with distal (p < 0.01), but not with proximal (p > 0.05), colonic transit. In contrast, after ES, stool output was determined by proximal colonic transit (p < 0.05) but not by transit through the distal colon (p > 0.05). CONCLUSION: The distal colon retains non absorbable fluid volumes extensively. In our model transit through the distal colon--but not the proximal colon--determined the time at which diarrhoea occurred. PMID- 9414975 TI - Efficacy and safety of the peripheral kappa agonist fedotozine versus placebo in the treatment of functional dyspepsia. AB - BACKGROUND: Peripheral kappa receptor agonists may provide a new therapeutic approach for the treatment of functional dyspepsia. AIMS: To evaluate, in a large multicentre trial, the use of the kappa receptor agonist fedotozine to improve symptoms associated with functional dyspepsia. METHODS: Two or more of the following persistent symptoms were required for inclusion: epigastric pain, early satiety, epigastric fullness or distension, nausea, vomiting, and a feeling of slow digestion. On completing a two week placebo washout, 271 patients were randomised into two groups to receive 30 mg fedotozine three times daily or placebo for six weeks under double blind conditions. RESULTS: The improvement in the overall intensity of dyspeptic symptoms (main efficacy criterion) was significantly more pronounced in the fedotozine group (p = 0.002) compared with placebo, as was epigastric pain (p = 0.004) and nausea (p = 0.01); the improvement in postprandial fullness was nearly significant (p = 0.052). Inability to finish a meal and slow digestion were unaffected. The patient global score, the average of the five individual symptoms, was notably ameliorated with fedotozine (p = 0.021). The safety of fedotozine was excellent. CONCLUSIONS: Fedotozine at 30 mg three times daily is safe and more effective than placebo for the relief of key symptoms associated with functional dyspepsia. PMID- 9414977 TI - Clinical characteristics of chronic idiopathic intestinal pseudo-obstruction in adults. AB - BACKGROUND: Chronic idiopathic intestinal pseudo-obstruction, a syndrome of ineffectual motility due to a primary disorder of enteric nerve or muscle, is rare. AIMS: To determine the clinical spectrum, underlying pathologies, response to treatments, and prognosis in a consecutive unselected group of patients. METHODS: Cross sectional study of all patients with clinical and radiological features of intestinal obstruction in the absence of organic obstruction, associated with dilated small intestine (with or without dilated large intestine), being actively managed in one tertiary referral centre at one time. RESULTS: Twenty patients (11 men and nine women, median age 43 years, range 22 67) fulfilled the diagnostic criteria. Median age at onset of symptoms was 17 years (range two weeks to 59 years). Two patients had an autosomally dominant inherited visceral myopathy. Major presenting symptoms were pain (80%), vomiting (75%), constipation (40%), and diarrhoea (20%). Eighteen patients required abdominal surgery, and a further patient had a full thickness rectal biopsy. The mean time interval from symptom onset to first operation was 5.8 years. Histology showed visceral myopathy in 13, visceral neuropathy in three, and was indeterminate in three. In the one other patient small bowel motility studies were suggestive of neuropathy. Two patients died within two years of symptom onset, one from generalised thrombosis and the other from an inflammatory myopathy. Of the remaining 18 patients, eight were nutritionally independent of supplements, two had gastrostomy or jejunostomy feeds, and eight were receiving home parenteral nutrition. Five patients were opiate dependent, only one patient had benefited from prokinetic drug therapy, and five patients required formal psychological intervention and support. CONCLUSIONS: In a referral setting visceral myopathy is the most common diagnosis in this heterogeneous syndrome, the course of the illness is usually prolonged, and prokinetic drug therapies are not usually helpful. Ongoing management problems include pain relief and nutritional support. PMID- 9414976 TI - Do patients with irritable bowel syndrome in primary care really differ from outpatients with irritable bowel syndrome? AB - BACKGROUND: Little is known about the comparability of outpatients with irritable bowel syndrome (IBS) and patients with IBS in primary care with regard to severity of complaints, perceived limitations, other aspects of the complaints, and sex differences. AIMS: To compare outpatients with IBS with primary care patients with IBS. PATIENTS: One hundred and nine patients with IBS were recruited from general practices in Amsterdam and 86 patients with IBS were recruited from the outpatient clinic of the Department of Internal Medicine of the University Hospital in Nijmegen. METHODS: Each patient completed a questionnaire on demographic variables, abdominal complaints, related complaints, and attributed causes of their abdominal complaints. The scores of the two groups were compared by univariate and multivariate analysis. RESULTS: The outpatient group contained significantly more men, reported more severe abdominal pain, more frequent complaints, more interference with daily activities, and a higher degree of avoidance of activities (p < 0.01) than the primary care group. When each sex was analysed separately, these differences remained for female (p < 0.01) but not for male patients. Outpatients were more likely to attribute their complaints to somatic causes (p < 0.01), whereas primary care patients were more likely to attribute their complaints to stress (p < 0.01) or their agitated way of life (p < 0.05). Multivariate analysis showed that a high severity score, a large number of additional complaints, and a low score on the stress attribution were important determinants for being in the outpatient group. CONCLUSIONS: Female outpatients consider their complaints to be more serious and interfering than do patients with IBS in primary care. Male outpatients were comparable to primary care patients with IBS. More research needs to be done into sex specific differences in IBS and into the factors that influence the decision to refer a patient with IBS. PMID- 9414979 TI - Sclerosing cholangitis, race and sex. AB - BACKGROUND: Primary sclerosing cholangitis develops in 3-10% of patients with ulcerative colitis, and may be associated with an increased cancer risk. Ulcerative colitis is probably less common in people of African origin than in populations of European descent. AIMS AND METHODS: To review the records of all patients under regular follow up for ulcerative colitis at St Bartholomew's Hospital (London, UK), a tertiary referral centre, prompted by discovering a cluster of cases with common features. RESULTS AND CONCLUSIONS: Among 166 patients with ulcerative colitis under regular follow up, only four (all women) are of African or Caribbean genetic origin, and three of these have developed sclerosing cholangitis within three years of presentation with colitis, compared with four of 162 patients of European or Asian descent (odds ratio 119, 95% confidence interval 8-3837; p = 0.0002). This cluster, which is not explained by common HLA DR or DQ type, suggests that Africans and Afro-Caribbeans, especially women, may be at increased risk of sclerosing cholangitis. This may reflect genetic influences on the development of enteric and hepatobiliary inflammatory disease. PMID- 9414978 TI - Increased incidence of biliary sludge and normal gall bladder contractility in patients with high spinal cord injury. AB - BACKGROUND: Patients with spinal cord injury (SCI) have an increased prevalence of gallstones. AIMS: To study prospectively the incidence of gallstones and gall bladder contractility in patients with SCI. PATIENTS AND METHODS: Thirty six consecutive patients with SCI were studied: 18 patients with SCI above thoracic 10 neuronal segment (> T10) and 18 patients with SCI below T10 (< T10). An equal number each of disease controls (multiple fractures) and healthy controls were also studied. All patients and controls underwent serial ultrasonography to detect development of gallstones and ultrasonographic measurement of gall bladder contractility. RESULTS: A significantly higher number (9/18) of patients with SCI > T10 developed biliary sludge compared with patients with SCI < T10 (2/18), disease controls (2/18), and healthy controls (1/18) (p < 0.05). No patient developed gallstones. The gall bladder fasting volume was significantly decreased in patients with SCI > T10 (20.56 ml; 95% confidence intervals (CI) 19.74 to 21.38) compared with that in patients with SCI < T10 (27.33 ml, 95% CI 26.17 to 28.49; p < 0.05), disease controls (27.92 ml, 95% CI 26.69 to 29.15; p < 0.05), and healthy controls (28.35 ml, 95% CI 27.25 to 29.45; p < 0.05). Gall bladder contractility was normal in patients with SCI as shown by normal gall bladder residual volume and emptying time. CONCLUSIONS: Patients with SCI above T10 have an increased incidence of biliary sludge and a decreased gall bladder fasting volume. Gall bladder contractility is, however, normal. PMID- 9414980 TI - Increased serum trypsinogen 2 and trypsin 2-alpha 1 antitrypsin complex values identify endoscopic retrograde cholangiopancreatography induced pancreatitis with high accuracy. AB - AIMS: To evaluate the clinical utility of two new tests for serum trypsinogen 2 and trypsin 2-alpha 1 antitrypsin complex (trypsin 2-AAT) in diagnosing and assessing the severity of acute pancreatitis (AP) induced by endoscopic retrograde cholangiopancreatography (ERCP). PATIENTS: Three hundred and eight consecutive patients undergoing ERCP at Helsinki University Central Hospital in 1994 and 1995. METHODS: Patients were followed prospectively for pancreatitis and clinical outcome. They were tested for serum trypsinogen 2, trypsin 2-AAT, and amylase in samples obtained before and one, six, and 24 hours after ERCP. RESULTS: Pancreatitis developed in 31 patients (10%). Their median serum trypsinogen 2 increased 26-fold to 1401 micrograms/l at six hours after the procedure and trypsin 2-AAT showed an 11-fold increase to 88 micrograms/l at 24 hours. The increase in both markers was stronger in severe than in mild pancreatitis, and in patients without pancreatitis there was no significant increase. Baseline trypsinogen 2 and trypsin 2-AAT concentrations were elevated in 29% and 32% of patients, respectively. The diagnostic accuracy of a threefold elevation over the baseline value was therefore analysed. The sensitivity and specificity of these parameters in the diagnosis of post-ERCP pancreatitis was 93% and 91%, respectively, for serum trypsinogen 2 at six hours after the examination, and 93% and 90%, for trypsin 2-AAT at 24 hours. CONCLUSIONS: Serum trypsinogen 2 and trypsin 2-AAT reflect pancreatic injury after ERCP. High concentrations are associated with severe pancreatic damage. The delayed increase in trypsin 2-AAT compared with trypsinogen 2 appears to reflect the pathophysiology of AP. A greater than threefold increase in trypsinogen 2 six hours after ERCP is an accurate indicator of pancreatitis. PMID- 9414981 TI - Hepatitis G virus infection in fulminant hepatic failure. AB - BACKGROUND: RNA sequences of the recently identified hepatitis GB virus C (HGBV C), also named hepatitis G virus (HGV), have been detected in patients with idiopathic fulminant hepatic failure (FHF) but the role of this agent in the disease remains controversial. AIMS: To investigate the presence and implications of HGV infection in a large series of Spanish patients with FHF. PATIENTS: Sixty eight patients with FHF, including 19 with idiopathic disease, were studied. In 28 cases, studies were performed before and after liver transplantation. For comparison 200 volunteer blood donors and 22 patients transplanted for chronic liver disease were also studied. METHODS: HGV RNA was measured in serum by reverse transcriptase polymerase chain reaction of the 5' non-coding region. RESULTS: Evidence of HGV infection was found in 3% (6/200) of blood donors and in 19% (13/68) of patients with FHF. HGV infection was more frequent in patients with hepatitis B (24%, 6/25) or hepatitis D (42%, 5/12), than in patients with idiopathic disease (11%, 2/19). Half of the patients with HGV infection used illicit intravenous drugs. Specific clinical features associated with HGV infection were not identified. A very high rate of infection with HGV was observed in patients who underwent liver transplantation, either for FHF (60%, 15/24) or chronic liver disease (45%, 9/20). CONCLUSIONS: In our geographical area, HGV infection is relatively frequent in FHF, but it does not seem to play a major role in idiopathic cases. PMID- 9414982 TI - The effect of plasma low density lipoprotein apheresis on the hepatic secretion of biliary lipids in humans. AB - BACKGROUND: The liver is a key organ in the metabolism of cholesterol in humans. It is the only organ by which substantial amounts of cholesterol are excreted from the body, either directly as free cholesterol into the bile or after conversion to bile acids. The major part of cholesterol synthesis in the body occurs in the liver. Cholesterol is also taken up by the liver from plasma lipoproteins. The relative contributions of newly synthesised cholesterol and plasma lipoprotein cholesterol to bile acid synthesis and biliary cholesterol secretion, respectively, are not known in detail. AIMS: To determine how a rapid lowering of plasma low density lipoprotein (LDL) and very low density lipoprotein (VLDL) cholesterol influences the biliary secretion rates of cholesterol and bile acids in patients with cholesterol gallstones and complete biliary drainage. In this model with a completely interrupted enterohepatic circulation, the secretion of bile acids equals the new synthesis of bile acids in the liver. PATIENTS: Eight patients with common bile duct stones of cholesterol type undergoing conventional cholecystectomy and choledocholithotomy. METHODS: At operation a balloon occludable Foley catheter attached to a T tube was inserted into the bile duct with the balloon placed just past the distal limb of the T tube. The T tube was allowed to drain the bile externally. One week after the operation the Foley catheter balloon was inflated, creating complete biliary drainage. Twelve hours following the inflation plasma LDL apheresis was carried out for two hours. Bile was collected for 15 minute periods starting one hour before the apheresis and ending two hours after its termination. During the collection of bile, plasma lipids were analysed on several occasions. RESULTS: The plasma level of LDL cholesterol decreased by 26% from (mean (SEM)) 2.19 (0.29) to 1.63 (0.17) mmol/l during the LDL apheresis while high density lipoprotein (HDL) cholesterol in plasma was unaffected. During LDL apheresis apolipoprotein B containing lipoproteins bind to the column, causing a significant decrease of not only plasma LDL but also of VLDL cholesterol. The secretion rate of bile acids decreased significantly by 31% from 131 (38) to 90 (16) mumol/15 minutes (p = 0.045). The output of phospholipids also decreased by 19%. The biliary secretion rate of cholesterol was not, however, affected by the plasma LDL apheresis. CONCLUSIONS: The results suggest that, in patients with cholesterol gallstones and complete biliary drainage, lowering of plasma LDL and VLDL cholesterol reduces the biliary secretion rate--synthesis--of bile acids without affecting the biliary secretion rate of cholesterol. PMID- 9414983 TI - Pancreatitis induced by codeine: a case report with positive rechallenge. AB - A woman who developed acute pancreatitis following ingestion of low dose codeine, with positive rechallenge, is described. As this is the first case report of pancreatitis being induced solely by codeine, this side effect must be rare in view of the widespread consumption of this drug. PMID- 9414985 TI - The columnar lined oesophagus: a riddle wrapped in a mystery inside an enigma. PMID- 9414984 TI - Peyer's patch organogenesis--cytokines rule, OK? AB - Targeted inactivation of genes in the tumor necrosis factor (TNF)/lymphotoxin (LT) ligand and receptor system has recently revealed essential roles forthese molecules in lymphoid tissue development and organization. Lymphotoxin-alpha beta (LT alpha beta)/lymphotoxin-beta receptor (LT beta-R) signaling is critical for the organogenesis of lymph nodes and Peyer's patches and for the structural compartmentalization of the splenic white pulp into distinct B and T cell areas and marginal zones. Moreover, an essential role has been demonstrated for TNF/p55 tumor necrosis factor receptor (p55TNF-R) signaling in the formation of splenic B lymphocyte follicles, follicular dendritic cell networks, and germinal centers. In contrast to a previously described essential role for the p55TNF-R in Peyer's patch organogenesis, we show in this report that Peyer's patches are present in both TNF and p55TNF-R knockout mice, demonstrating that these molecules are not essential for the organogenesis of this lymphoid organ. Furthermore, we show that in the absence of TNF/p55TNF-R signaling, lymphocytes segregate normally into T and B cell areas and a normal content and localization of dendritic cells is observed in both lymph nodes and Peyer's patches. However, although B cells are found to home normally within Peyer's patches and in the outer cortex area of lymph nodes, organized follicular structures and follicular dendritic cell networks fail to form. These results show that in contrast to LT alpha beta signaling, TNF signaling through the p55TNF-R is not essential for lymphoid organogenesis but rather for interactions that determine the cellular and structural organization of B cell follicles in all secondary lymphoid tissues. PMID- 9414986 TI - Why do we hiccup? PMID- 9414987 TI - The duodenum: a conduit or a pump? PMID- 9414988 TI - Assessing food intolerance: don't lose control. PMID- 9414989 TI - Visceral analgesics and functional dyspepsia: have we found the Holy Grail? PMID- 9414990 TI - Treating diarrhoea: what might the pituitary offer? PMID- 9414991 TI - Homing-in on the origin of biliary steroids. PMID- 9414992 TI - Defining treatment resistance in the irritable bowel syndrome. PMID- 9414993 TI - Are Down syndrome and coeliac disease associated? PMID- 9414994 TI - Helisal saliva assay. PMID- 9414995 TI - Delayed protection against ventricular arrhythmias by cardiac pacing. PMID- 9414997 TI - Possible uses of gene therapy in reducing coronary restenosis. PMID- 9414996 TI - Karl Freiherr von Rokitansky (1804-78). PMID- 9414998 TI - The origin of symptoms in chronic heart failure. PMID- 9414999 TI - The role of exercise training in chronic heart failure. PMID- 9415000 TI - Metabolic abnormality of calf skeletal muscle is improved by localised muscle training without changes in blood flow in chronic heart failure. AB - OBJECTIVE: To investigate whether localised skeletal muscle training, which does not have a great influence on the heart, improves abnormalities of calf muscle metabolism in patients with chronic heart failure. METHODS: Seven cardiac patients in New York Heart Association class II and III undertook a random order crossover trial. Training consisted of unilateral calf plantar flexion exercise. Before and after training, the patients' metabolic responses were examined during the calf exercise test with phosphorus-31 nuclear magnetic resonance spectroscopy (31P-MRS) and calf blood flow with plethysmography. The new Borg scale was employed as a subjective fatigue scale. RESULTS: In a constant load exercise test (70% of maximum load achieved during the incremental exercise), standardised phosphocreatine and intracellular pH decreased less after training (p < 0.05, repeated measures analysis of variance). The new Borg scale improved significantly after training (p < 0.05). Blood flow did not change significantly in either test. CONCLUSIONS: In patients with chronic heart failure, localised calf skeletal muscle training improved oxidative capacity without changes in calf blood flow. This training also improved the subjective fatigue scale. This training method may therefore alleviate leg fatigue experienced in daily activities. PMID- 9415001 TI - Beta blockade in congestive heart failure: persistent adverse haemodynamic effects during chronic treatment with subsequent doses. AB - OBJECTIVE: To determine whether the acute adverse haemodynamic effects of beta blockade in patients with congestive heart failure persist during chronic treatment. DESIGN: Sequential haemodynamic evaluation of heart failure patients at baseline and after three months of continuous treatment with the beta 1 selective antagonist metoprolol. SETTING: Cardiac care unit in university hospital. PATIENTS: 26 patients with moderate to severe congestive heart failure (New York Heart Association grade II to IV) and background treatment with digoxin, diuretics, and angiotensin converting enzyme inhibitors, and with a left ventricular ejection fraction < 25%. METHODS: Baseline variables included a six minute walk, maximum oxygen consumption, and right heart catheterisation. All patients received metoprolol 6.25 mg orally twice daily initially and the dose was gradually increased to a target of 50 mg twice daily. Haemodynamic measurements were repeated after three months of treatment, both before (trough) and after drug readministration. RESULTS: Long term metoprolol had functional, exercise, and haemodynamic benefits. It produced decreases in heart rate, pulmonary capillary wedge pressure, and systemic vascular resistance, and increases in cardiac index, stroke volume index, and stroke work index. However, when full dose metoprolol was readministered during chronic treatment, there was a reduction in cardiac index (from 2.8 (SD 0.46) to 2.3 (0.38) l/min/m2, p << 0.001) and stroke work index (from 31.4 (11.1) to 26.6 (10.0) g.m/m2, p < 0.001) and an increase in systemic vascular resistance (from 943 (192) to 1160 (219) dyn.s.cm-5, p << 0.001). CONCLUSIONS: Adverse haemodynamic effects of beta blockers in heart failure persist during chronic treatment, as shown by worsening haemodynamic indices with subsequent doses. PMID- 9415002 TI - Dietary determinants of ischaemic heart disease in health conscious individuals. AB - OBJECTIVE: To investigate dietary determinants of ischaemic heart disease (IHD) in health conscious individuals to explain the reduced risk in vegetarians, and to examine the relation between IHD and body mass index (BMI) within the normal range. DESIGN: Prospective observation of vegetarians, semi-vegetarians, and meat eaters for whom baseline dietary data, reported weight and height information, social class, and smoking habits were recorded. SUBJECTS: 10,802 men and women in the UK aged between 16 and 79, mean duration of follow up 13.3 years. MAIN OUTCOME MEASURES: Death rate rations for IHD and total mortality in relation to dietary and other characteristics recorded at recruitment (reference category death rate = 100). RESULTS: IHD mortality was less than half that expected from the experience reported for all of England and Wales. An increase in mortality for IHD was observed with increasing intakes of total and saturated animal fat and dietary cholesterol-death rate ratios in the third tertile compared with the first tertile: 329, 95% confidence interval (CI) 150 to 721; 277, 95% CI 125 to 613; 353, 95% CI 157 to 796, respectively. No protective effects were observed for dietary fibre, fish or alcohol. Within the study, death rate ratios were increased among those in the upper half of the normal BMI range (22.5 to < 25) and those who were overweight (BMI > or = 25) compared with those with BMI 20 to < 22.5. CONCLUSIONS: In these relatively health conscious individuals the deleterious effects of saturated animal fat and dietary cholesterol appear to be more important in the aetiology of IHD than the protective effect of dietary fibre. Reduced intakes of saturated animal fat and cholesterol may explain the lower rates of IHD among vegetarians compared with meat eaters. Increasing BMI within the normal range is associated with increased risk of IHD. The results have important public health implications. PMID- 9415003 TI - Reduction in treatment delay by paramedic ECG diagnosis of myocardial infarction with direct CCU admission. AB - OBJECTIVES: To establish the feasibility of training paramedics of diagnose acute myocardial infarction by ECG before hospital admission and whether direct paramedic coronary care admission, arranged by very high frequency (VHF) radio communication with the coronary care unit (CCU), would reduce delay of thrombolysis treatment. DESIGN: Prospective controlled study. SETTING: District general hospital CCU and a local district ambulance paramedic service. PATIENTS: 124 patients with ECG evidence of myocardial infarction or ischaemia admitted directly to the CCU by the paramedic service were compared with 123 patients admitted by the emergency department and subsequently transferred to the CCU. MAIN OUTCOME MEASURES: ECG diagnostic accuracy by paramedics, and interval durations for CCU admission and thrombolysis. RESULTS: ECG diagnostic accuracy by the paramedics was 87.5% in the training phase and 92% in admission. The total call to thrombolysis interval was reduced from 154 to 93 minutes and the "door to needle" interval was reduced from 97 to 37 minutes. CONCLUSIONS: Trained paramedics can reliably diagnose myocardial infarction by ECG. The use of a direct admission procedure, by a VHF radio link to the CCU, substantially reduces the time interval for thrombolytic treatment after acute myocardial infarction. PMID- 9415004 TI - Direct admission to the coronary care unit by the ambulance service for patients with suspected myocardial infarction. AB - BACKGROUND: Direct access to the coronary care unit (CCU) for general practitioner (GP) referred cases of suspected acute myocardial infarction (AMI) (fast track admission) substantially reduces the time to thrombolysis. Until now, this policy has been confined to GP referrals. OBJECTIVES: To determine the time taken to admission to CCU under the fast track policy (ambulance referrals and GP referrals) and the time taken to start administration of thrombolytics (ambulance referrals, GP referrals, and accident and emergency referrals). METHODS: Fast track admission policy was extended to include referrals from ambulance personnel who respond to emergency service calls. Ambulance personnel referred cases were also examined to see if they were referred appropriately to the CCU. RESULTS: 100 ambulance personnel referrals and 260 GP referrals to CCU with chest pain were studied. Forty accident and emergency referrals who had AMI requiring thrombolysis were also studied. In the ambulance referred group the time to admission from phone call was a median of 10 minutes (range 2 to 45), a saving of 30 minutes compared with GP referrals (median 40 minutes, range 2 to 217). The median diagnostic electrocardiogram (ECG) to thrombolysis time was longer in the accident and emergency referrals with AMI than either ambulance referrals or GP referrals admitted under the fast track policy. Diagnostic ECG to thrombolysis time: accident and emergency 50 minutes (range 15 to 385); ambulance referrals median 33 minutes (range 6 to 69); GP referrals median 29.5 minutes (range 5 to 110 minutes); (p = 0.056 accident and emergency compared with ambulance referrals, p < 0.002 accident and emergency compared with GP referrals). Of 100 ambulance referrals 52 patients exhibited symptoms suggestive of ischaemic heart disease (confirmed AMI, unstable angina, and angina) and a further 18 patients were required to stay in CCU for other cardiac problems. Thus a total of 70 (70%) were considered appropriate compared with 155 of 260 (55.8%) GP referred cases. CONCLUSIONS: Extending the fast track admission policy to ambulance personnel reduces delay to admission for patients with suspected MI without adversely affecting the appropriateness of admissions. PMID- 9415005 TI - Change in ST segment elevation 60 minutes after thrombolytic initiation predicts clinical outcome as accurately as later electrocardiographic changes. AB - OBJECTIVE: To compare prospectively the prognostic accuracy of a 50% decrease in ST segment elevation on standard 12-lead electrocardiograms (ECGs) recorded at 60, 90, and 180 minutes after thrombolysis initiation in acute myocardial infarction. DESIGN: Consecutive sample prospective cohort study. SETTING: A single coronary care unit in the north of England. PATIENTS: 190 consecutive patients receiving thrombolysis for first acute myocardial infarction. INTERVENTIONS: Thrombolysis at baseline. MAIN OUTCOME MEASURES: Cardiac mortality and left ventricular size and function assessed 36 days later. RESULTS: Failure of ST segment elevation to resolve by 50% in the single lead of maximum ST elevation or the sum ST elevation of all infarct related ECG leads at each of the times studied was associated with a significantly higher mortality, larger left ventricular volume, and lower ejection fraction. There was some variation according to infarct site with only the 60 minute ECG predicting mortality after inferior myocardial infarction and only in anterior myocardial infarction was persistent ST elevation associated with worse left ventricular function. The analysis of the lead of maximum ST elevation at 60 minutes from thrombolysis performed as well as later ECGs in receiver operating characteristic curves for predicting clinical outcome. CONCLUSION: The standard 12-lead ECG at 60 minutes predicts clinical outcome as accurately as later ECGs after thrombolysis for first acute myocardial infarction. PMID- 9415006 TI - Cardiac valve calcification: characteristics of patients with calcification of the mitral annulus or aortic valve. AB - AIMS: To determine whether mitral annular calcification and aortic valve calcification, with or without stenosis, are expressions of atherosclerotic disease. METHODS: The incidence of atherosclerotic risk factors was analysed in patients with mitral annular calcification and aortic valve calcification and in control patients from a prospective echocardiographic database of 8160 consecutive patients; 657 patients (8%) were identified with mitral annular calcification and 815 (9%) with a calcified aortic valve, of whom 515 (6.3%) had stenosis with a minimal aortic valve gradient of 16 mm Hg. In these patients, cardiac and vascular risk factors were compared with 568 control patients using multiple logistic regression analysis. RESULTS: Age (odds ratio (OR) varying from 5.78 to 104, depending on age class), female sex (OR 1.75), hypertension (OR 2.38), diabetes mellitus (OR 2.85), and hypercholesterolaemia (OR 2.95) were strongly and significantly associated with aortic valve calcification without stenosis, as were age (OR varying from 8.82 to 67, depending on age class), female sex (OR 2.22), hypertension (OR 2.72), diabetes mellitus (OR 2.49), and hypercholesterolaemia (OR 2.86) with mitral annular calcification. Age (OR varying from 1.11 to 7.7), hypertension (OR 1.91), and hypercholesterolaemia (OR 2.55) were strongly and significantly associated with stenotic aortic valve calcification. CONCLUSIONS: Mitral annular calcification and stenotic or non stenotic aortic valve calcification have a high incidence of atherosclerotic risk factors, suggesting they should be considered as manifestations of generalised atherosclerosis. PMID- 9415008 TI - Right ventricular dysfunction during coronary artery occlusion: pressure-volume analysis using conductance catheters during coronary angioplasty. AB - OBJECTIVE: To study the effects of coronary artery occlusion on the pressure volume relations of the right ventricle. DESIGN: Right ventricular pressure volume cycles were studied using conductance catheters and micromanometers in 19 subjects undergoing coronary angioplasty in a tertiary referral cardiac centre. RESULTS: Catheter occlusions of either the left anterior descending coronary artery or the right coronary artery were associated with a decline in stroke work (mean change (SD): left-13.3 (15.8)%, p = 0.008; right -13.5(16.5)%, p = 0.04). Two patterns of change were evident: an upward shift usually associated with occlusion in the left coronary artery, and a rightward shift in the right coronary artery. In the former there was an increase in maximum ventricular volume (mean change: 3.0(2.7)%, p = 0.004) and in minimum ventricular volume (mean change: 2.3(2.7)%, p = 0.01) and a fall in peak pressure (mean change: -4.8 (5.1)%, p = 0.04). In the latter there was an increase in peak pressure (mean change 9.9(16.3)%, p = 0.04) and an increase in minimum ventricular volume (mean change 3.7(5.0)%, p = 0.02) leading to a fall in stroke volume (mean change 13.3(15.8)%, p = 0.008). CONCLUSIONS: Occlusion of the left anterior descending coronary artery or the right coronary artery is associated with a decline in right ventricular work. However, different patterns of change in indices of preload and afterload lead to different effects on overall right ventricular pump function. PMID- 9415007 TI - A case-control investigation of the relation between hyperlipidaemia and calcific aortic valve stenosis. AB - OBJECTIVE: To investigate the relation of hyperlipidaemia to calcific aortic valve stenosis. DESIGN: A case-control study designed to detect a clinically relevant difference in the fasting plasma concentrations of total cholesterol between the groups at the 5% level with a power of 90%. Predefined subgroup analyses were based on presence of significant coronary disease and valve morphology (that is, bicuspid or tricuspid). SETTING: A district general hospital. SUBJECTS: 20 patients with severe calcific aortic stenosis and 20 controls. RESULTS: Mean (SD) fasting plasma total cholesterol in patients with aortic stenosis was 0.79 (1.50) mmol/l greater than in the controls (p = 0.029). The magnitude of differences between patients with aortic stenosis and controls was similar whether the patients had coronary artery disease (0.78 (1.73) mmol/l) or not (0.80 (1.37) mmol/l). The presence of a stenosed tricuspid aortic valve was associated with a significant increase in plasma cholesterol (1.70 (0.87) mmol/l, p = 0.012). For bicuspid valves the degree of elevation of plasma cholesterol was less and not statistically significant. CONCLUSIONS: Calcific aortic stenosis is associated with hypercholesterolaemia, especially when the valve is tricuspid. Further studies are necessary to confirm that the relation is causal. This finding may have implications for measures to prevent the most common cause of cardiac valve replacement in the developed world. PMID- 9415009 TI - Changes in pulmonary artery size before and after total cavopulmonary connection. AB - OBJECTIVE: To assess changes in size of the central pulmonary arteries following a total cavopulmonary connection (TCPC). DESIGN: A retrospective analysis of the angiographic diameters of the central pulmonary arteries, expressed as z scores, in infancy before the TCPC and 3.5 (0.9) years (mean (SD)) later. Analysis of the relation between the pulmonary arteriolar resistance and the z scores at follow up. SETTING: Tertiary referral centre. PATIENTS: 32 patients who had TCPC from February 1990 to July 1993. RESULTS: The patients were divided into two groups (n = 16) depending on their preoperative flow ratio: group I, Qp/Qs < or = 1; group II, Qp/Qs > 1. At the initial study in infancy the mean z scores in group I were 6.0 for the right pulmonary artery (RPA) and -9.6 for the left pulmonary artery (LPA); in group II the respective values were -2.7 and -3.0. Before the TCPC the values increased to 0.5 (RPA) and -0.5 (LPA) in group I, and to 8.8 (RPA) and 8.2 (LPA) in group II. At follow up the z scores decreased to -2.4 (RPA) and -4.9 (LPA) in group I, and to 2.2 (RPA) and -0.7 (LPA) in group II. The changes in pulmonary artery diameters were significant for both groups (p < 0.02). Following the TCPC, no significant difference in pulmonary arteriolar resistance index was found between patients with relatively small pulmonary arteries (z score RPA + LPA < or = 0) and those with relatively large pulmonary arteries (z score RPA + LPA > 0). CONCLUSIONS: Creation of a TCPC results in a significant reduction in size of the central pulmonary arteries. At a mean interval of 3.5 years following the TCPC, however, there was no significant difference in pulmonary arteriolar resistance index between patients with smaller and larger central pulmonary arteries. PMID- 9415010 TI - Images in cardiology. Left atrial dissection after mitral valve reconstruction. PMID- 9415012 TI - Delayed improvement of autonomic nervous abnormality after the Maze procedure: time and frequency domain analysis of heart rate variability using 24 hour Holter monitoring. AB - OBJECTIVE: To analyse heart rate variability in patients with atrial fibrillation after the Maze procedure, to investigate whether the procedure damages the cardiac autonomic fibres supplying the sinus node. DESIGN AND PATIENTS: Time and frequency domain analyses of RR variability were performed using 24 hour Holter monitoring one month after surgery in 12 patients with atrial fibrillation who underwent the Maze procedure (Maze group) and in seven patients who underwent cardiac surgery without the Maze procedure (control group). Mean RR intervals (mRR) and the standard deviation of successive RR intervals (SDRR) were determined by time domain analysis, and high frequency (HF), low frequency (LF), and total power (TP) spectral components of RR variability were calculated by frequency domain analysis. Holter monitoring was also performed at six and 12 months after cardiac surgery in the Maze group. RESULTS: Circadian variation (mean (SD)) in mRR (daytime to night time difference: 119 (60) v 302 (143) ms), SDRR (daytime: 8.4 (3.3) v 37.0 (12.0) ms), TP (daytime: 46.7 (16.0) v 171.8 (30.4) ms), HF (daytime: 19.6 (9.9) v 36.7 (7.1) ms2), and LF/HF (daytime: 0.31 (0.07) v 1.18 (0.46)) was decreased in the Maze group at one month compared with the control group (p < 0.01), but showed improvement at six and 12 months (p < 0.05). CONCLUSIONS: Surgery combined with the Maze procedure markedly suppressed the circadian variation of heart rate over a 24 hour period within one month after surgery, mainly because of damage to the innervation of the sinus node. However, at six and 12 months there was restoration of circadian variation, probably as the result of reinnervation of the sinus node. PMID- 9415015 TI - Right atrial myxoma mistaken for recurrent pulmonary thromboembolism. AB - A 69 year old man was admitted for investigation of right sided pleuritic chest pain and dyspnoea, both of which began suddenly four days before admission. Acute pulmonary embolism was diagnosed. Six months after discharge while on warfarin he died. Necropsy found a 50 mm diameter myxoid tumour arising on the right atrial side of the interatrial septum. This lesion may have been discovered earlier by echocardiography although there were no clear indications for this investigation. Presentation was that of recurrent pulmonary embolism with no obvious source or cause of thrombosis. Patients who are thought to have idiopathic pulmonary embolism should undergo early echocardiography to exclude the rare but treatable diseases of the right heart that may be responsible. PMID- 9415013 TI - Measuring serum aminoterminal type III procollagen peptide, 7S domain of type IV collagen, and cardiac troponin T in patients with idiopathic dilated cardiomyopathy and secondary cardiomyopathy. AB - OBJECTIVE: To identify new prognostic indicators in idiopathic dilated cardiomyopathy (DCM) and secondary cardiomyopathy. DESIGN AND PATIENTS: Serum concentrations of aminoterminal propeptides of type III procollagen and the 7S domain of type IV collagen (7S collagen)--which have recently been used as indicators of collagen matrix turnover in other diseases--and of cardiac troponin T were measured in 17 consecutive patients with DCM and in four patients with secondary cardiomyopathy (one associated with hyperthyroidism, two with chronic renal failure, one with amyloidosis), confirmed by endomyocardial biopsy. The correlation of these variables with short term prognosis was then assessed prospectively. RESULTS: 11 of the patients were positive for type III procollagen, 7S collagen, or troponin T even though their creatine kinase concentrations were within the normal range. These patients had a poor short term prognosis (p < 0.001). CONCLUSIONS: Within the DCM and secondary cardiomyopathy groups, there was a subgroup of patients with raised concentrations of serum collagen and troponin T, for whom short term prognosis was poor. Although it is unclear whether these serum peptide levels reflect ongoing myocyte degeneration and interstitial fibrosis, they may serve as useful new prognostic indicators for cardiomyopathy. PMID- 9415016 TI - Successful thrombolysis of right atrial and ventricle thrombi in a patient with peripartum cardiomyopathy and extensive thromboembolism. PMID- 9415014 TI - Human stress cardiomyopathy mimicking acute myocardial syndrome. AB - Two cases of transient acute cardiomyopathy occurring in the immediate aftermath of intense emotional stress and without any identified aetiology are described. These two cases reports, mimicking cases of acute cardiomyopathy described in patients with pheochromocytoma, suggest the possibility in man of acute catecholamine induced cardiomyopathy related to major emotional stress alone, a phenomenon so far reported only in animal experimental models. PMID- 9415011 TI - Bradykinin induced dilatation of human epicardial and resistance coronary arteries in vivo: effect of inhibition of nitric oxide synthesis. AB - OBJECTIVE: To clarify whether endothelium derived nitric oxide contributes to exogenous bradykinin induced dilatation of human epicardial and resistance coronary arteries in vivo. DESIGN: Quantitative coronary angiography and Doppler flow velocity measurements were used to determine the effects of the nitric oxide synthesis inhibitor, NG-monomethyl-L-arginine (L-NMMA), on bradykinin induced dilatation of the epicardial and resistance coronary arteries. SETTING: Hiroshima University Hospital. PATIENTS: 20 patients (16 men and four women, mean (SD) age 56 (9) years) with angiographically normal smooth epicardial coronary arteries. INTERVENTIONS: Serial infusions of bradykinin (0.5, 1.5, and 2.5 micrograms/min) were given into the left coronary ostium before and after L-NMMA infusion (60 mumol/min). MAIN OUTCOME MEASURES: Epicardial coronary diameter, coronary blood flow, and coronary vascular resistance. RESULTS: Bradykinin-induced epicardial coronary vasodilatation after L-NMMA (dilatation by 2.5 micrograms/min, 3.8(1.4)% in the proximal and 5.9(1.8)% in the distal segments, mean (SEM)) was less (p < 0.001, respectively) than before L-NMMA (11.7(2.5)% and 15.1(2.0)%, respectively). In contrast, L-NMMA did not affect the bradykinin induced increase in coronary blood flow and decrease in coronary vascular resistance. CONCLUSIONS: Endothelium derived nitric oxide contributes to bradykinin induced dilatation of epicardial coronary arteries, but may be less important in coronary resistance vasodilatation. PMID- 9415017 TI - Successful thrombolysis with intracoronary administration of tissue plasminogen activator in an infant with Kawasaki disease. PMID- 9415018 TI - The changing interface between district hospital cardiology and the major cardiac centres. British Cardiac Society, with the Royal College of Physicians of London, the Royal College of Physicians of Edinburgh, and the Royal College of Physicians and Surgeons of Glasgow. AB - The national priority for reducing mortality and morbidity from cardiovascular disease, the resulting expansion in the number of consultant cardiologists, and the reforms of the National Health Service have produced significant changes in delivery of care for cardiac patients and in the relations between district general hospitals (DGH) and the old regional cardiac centres. 1.2 The British Cardiac Society, the Medical Royal Colleges of Physicians of London and Edinburgh, and the Royal College of Physicians and Surgeons of Glasgow established a working group to make recommendations on the most appropriate evolution of these changes to secure high quality care in a cost-effective and professionally rewarding environment. The principal conclusions of the working group were: i) The establishment of new cardiac catheterisation laboratories in DGHs remote from a major cardiac centre should be encouraged provided the workload is adequate to ensure efficient use of the facility. ii) Cardiologists working in districts close to a major centre should be encouraged to catheterise their patients at the centre. iii) Close liaison of the district cardiologist with a cardiac surgeon and interventionist is vitally important. iv) The centres will be required to provide tertiary care for emergency and urgent cases from their traditional catchment area, specialised expertise for the management of rare and difficult cases, and angioplasty. Some centres will also offer complex electrophysiology, and ablation techniques. v) The centres must also provide routine cardiology services for their local district, facilities for cardiac catheterisation for DGH cardiologists, and training for doctors, nurses, technicians, and radiographers. vi) Some centres will be linked with paediatric cardiology and paediatric cardiac surgical units. vii) District cardiac centres will be required to provide a full non-invasive diagnostic service and emergency care for patients referred by general practitioners and hospital colleagues as well as facilities for preventative and rehabilitation cardiology. Arrangements for invasive investigation and treatment of their patients will vary according mainly to the distance from the major centre. viii) Both the major centres and the district cardiac units should participate in training and research. PMID- 9415019 TI - Hyperhomocysteinaemia, Helicobacter pylori, and coronary heart disease. PMID- 9415020 TI - Hyperhomocysteinaemia, Helicobacter pylori, and coronary heart disease. PMID- 9415021 TI - T-lymphocyte activation and the cellular form of the prion protein. AB - The transmissible spongiform encephalopathies are neurodegenerative disorders which include Creutzfeldt-Jakob disease in humans, and scrapie and bovine spongiform encephalopathy in animals. A major component of the infectious agent responsible for these diseases is considered to be a post-translationally modified form of a host-encoded glycoprotein PrPc, termed PrPSc. While PrPc is abundantly expressed in tissues of the central nervous system (CNS), little is known about its normal function. The expression of PrPc is not restricted to the CNS, as this protein can also be detected in the lymphoid tissues of mice and sheep. In this report we demonstrate that resting murine splenic lymphocytes express PrPc protein on their cell membranes. Furthermore, expression of PrPc was significantly enhanced following in vitro stimulation with the non-specific T cell mitogen concanavalin A (Con A). Genetically engineered mice with an inactive PrPc gene (PrP-/- mice), were utilized to investigate the involvement of PrPc in lymphocyte activation. Experiments revealed that the Con A-induced proliferation of lymphocytes from PrP-/- mice was significantly reduced to approximately 50-80% that of wild-type (PrP+/+) mice 48 hr post-stimulation. These findings demonstrate an important role for PrPc in extra-neuronal tissues and suggest that PrPc is a lymphocyte surface molecule that participates in T-cell activation. PMID- 9415022 TI - Modulation in vitro of human natural cytotoxicity, lymphocyte proliferative response to mitogens and cytokine production by essential fatty acids. AB - Essential fatty acids (EFA) have been shown in animal studies to have a differential effect on various aspects of immune reactivity. However, there have been few studies in humans. Therefore, we elected to investigate the effects of a variety of EFA [gamma-linolenic acid (GLA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA)] in vitro on human blood lymphocyte reactivity, cytokine secretion and natural cytotoxicity. The proliferative response to polyclonal mitogens (phytohaemagglutinin, pokeweed mitogen, concanavalin A), as measured by [3H]thymidine incorporation into newly synthesized lymphocytes, was inhibited (P < 0.05) by all EFAs tested, in a dose-dependent manner (3-15 micrograms/ml). The greatest inhibition of proliferation was caused by EPA and DHA. Similarly, EPA, DHA and GLA significantly reduced cytotoxic activity [expressed as lytic units, using 51 chromium-release assays natural killer (NK) (K562 cells) and lymphokine-activated (LAK) (Daudi cells) cells] (P < 0.05) in a concentration-dependent manner (5-50 micrograms/ml), without affecting cell viability. EPA and DHA exhibited greater suppression than GLA. Furthermore, the inhibition of cell proliferation and suppression of natural cytotoxicity was associated with marked decrease in cytokine [interleukin-1 (IL-1), IL-2, tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma)] production in vitro. Our findings demonstrate that EFAs (GLA, EPA, DHA) have the potential to inhibit significantly various aspects of human lymphocyte cell-mediated and humoral immune reactivities. PMID- 9415023 TI - Cellular distribution of a natural killer cell tumour recognition-related surface antigen in purified human lymphocytes. AB - Natural killer (NK) cells are large granular lymphocytes capable of human leucocyte antigen (HLA) unrestricted killing of tumour cells. A putative NK cell tumour-recognition molecule (NK-TR) was previously isolated and cloned. The predicted primary structure of the NK-TR revealed that the amino terminus of the protein shared high homology with cyclophilin proteins. In this study, we used rabbit antibodies directed against synthetic peptides corresponding to amino acids 476-497 of the NK-TR protein, to examine the expression of the NK-TR antigen in freshly purified human lymphocytes. Cell-surface staining experiments using these peptide antibodies indicated the presence of the NK-TR protein on the surface of human CD3+ T-cell populations purified from peripheral blood. There were individual donor differences in the levels of cell-surface expression of this antigen ranging from 35 to 90% in T lymphocytes and, NK cells purified from different healthy volunteers. The immunoreactivity of our peptide antibodies in immunoprecipitation showed that the NK-TR-related protein expressed in purified T cells is similar to that expressed in NK cells in terms of its electrophoretic mobility. Cell-surface staining experiments using the peptide antibodies revealed that the NK-TR-related protein is more abundantly expressed on the surface of purified T cells compared with NK cells. Northern blot analysis of the mRNA species transcribed in human lymphocytes revealed abundant expression of NK-TR specific mRNA species in purified T cells. Furthermore, another mRNA species smaller than 7 kb was detected in both NK and T-cell populations of lymphocytes freshly isolated from peripheral blood. Expression at the cell surface of a cyclophilin-homologous protein in purified human T lymphocytes may indicate another function for the reported NK-TR protein, that is, distinct from tumour cell recognition and cytosis. PMID- 9415024 TI - Later development of Fas ligand-mediated cytotoxicity as compared with granule mediated cytotoxicity during the maturation of natural killer cells. AB - We classified CD56+ CD3- natural killer (NK) cells into CD2- CD56dim (CD2- NK), CD2+ CD56dim (CD2+ NK) and CD2+ CD56bright populations, and investigated mainly functional differences between the former two populations. CD2- and CD2+ NK cells were the same in their morphology and several surface molecules except for CD2. The percentages of CD2- NK cells in total NK cells were higher in the cord blood and marrow than in the peripheral blood of adults or children. Freshly isolated CD2- NK cells had CD2 in the cytoplasm, and gradually expressed it on the surface upon incubation with interleukin-2 (IL-2). These results demonstrated that CD2 is an antigen which appears on the surface during the maturation of NK cells. The granule-mediated cytotoxicities, which are mainly performed by the perforin molecule, of CD2+ NK cells against K562 and Daudi cells were higher than those of CD2- NK cells, and they were inhibited to the levels of CD2- NK cells by the addition of a blocking anti-CD2 monoclonal antibody (mAb). Fas ligand (FasL) mRNA was expressed in freshly isolated CD2+ NK cells but not in the CD2- NK cells. Neither freshly isolated NK populations showed FasL-mediated cytotoxicity, and only CD2+ NK cells lysed Fas-transfected targets after the 24-hr incubation with IL-2. Based on these results, CD2- NK cells have already developed granule mediated cytotoxicity equal to that of CD2+ NK cells except for the CD2 associated activity, but they, unlike CD2+ NK cells, totally lack FasL-mediated cytotoxicity. These findings suggest that FasL-mediated cytotoxicity may be acquired at more mature stages of NK-cell maturation than granule-mediated cytotoxicity. PMID- 9415025 TI - Induction of selective anergy by toxic shock syndrome toxin-1 in CD8+ T cells. AB - Interferon-gamma (IFN-gamma) mRNA expression of murine splenic CD8+ T cells in response to toxic shock syndrome toxin-1 (TSST-1) was examined in vitro. In primary response to TSST-1, purified CD8+ cells expressed a low frequency of IFN gamma mRNA and produced very little interleukin-2 (IL-2) compared with unseparated lymphocytes and purified CD4+ cells. The addition of IL-2 to the cell culture enhanced IFN-gamma mRNA expression in the CD8+ population. Upon restimulation with TSST-1, no IL-2 production and hardly any IFN-gamma mRNA expression were observed in the purified CD8+ lymphoblasts. Exogenously added IL 2 did not increase the IFN-gamma mRNA expression in purified CD8+ blasts exposed to TSST-1. In contrast, stimulation with concanavalin A induced a high frequency of IFN-gamma mRNA in purified CD8+ blasts. Moreover, TSST-1 induced strong IFN gamma mRNA expression in unseparated T lymphoblasts and purified CD4+ populations. Interestingly, a combined immunocytochemistry and in situ hybridization technique showed a high frequency of CD8+ IFN-gamma mRNA-expressing cells in an unseparated blast population. These results demonstrate that TSST-1 induces a selective anergy within the purified CD8- T-cell compartment and that the IFN-gamma mRNA expression in CD8+ T blasts can be achieved by stimulation of unseparated T cells with TSST-1. PMID- 9415026 TI - Both CD4+ and CD8+ human lymphocytes are activated and proliferate in response to Cryptococcus neoformans. AB - The current studies were performed to determine the contribution of T-cell subsets to lymphocyte proliferation in response to Cryptococcus neoformans, the most common invasive mycosis in acquired immune deficiency syndrome. We demonstrate for the first time that both human CD4 and CD8 cells are activated in response to C. neoformans. Both CD4 and CD8 cells express interleukin-2 receptor alpha (IL-2R alpha) and transferrin receptor and proliferate in response to C, neoformans, however proliferation of CD8 cells was dependent upon CD4 cells. The requirement for CD4 cells was complex, since CD8 enriched cells failed to express mRNA for IL-2, suggesting that CD4-dependent IL-2 production was required for CD8 cell proliferation. However, IL-2 was not sufficient to restore CD8-cell proliferation. These studies provide experimental evidence in humans to support the clinical impression that CD4 cells are important in cryptococcosis, and suggest that the appropriate CD4-derived signals could allow CD8 cells to assist in host defence. PMID- 9415027 TI - Identification of nicotinic acetylcholine receptors on lymphocytes in the periphery as well as thymus in mice. AB - The existence of nicotinic acetylcholine receptors (nAChR) on lymphocytes remains controversial. We attempted to show the existence of nAChR on murine lymphocytes. The intraperitoneal injection of nicotine induced the lymphocytosis in the spleen on day 3. Although freshly isolated lymphocytes bound small quantities of fluorescein isothiocyanate (FITC)-conjugated alpha-bungarotoxin (alpha BuTx), they began to bind alpha BuTx after incubation in medium. In contrast to granulocytes, various lymphocyte subsets obtained from various lymphoid organs were found to bind alpha BuTx. Affinity purification of alpha BuTx-binding protein revealed that lymphocytes expressed the same nAChR molecules as those of muscle. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis showed that lymphocytes expressed the alpha-subunit mRNA of nAChR. These results suggest that lymphocytes carry nAChR on the surface and are stimulated directly via their nAChR by parasympathetic nerve stimuli. PMID- 9415028 TI - Apoptosis with FasL+ cell infiltration in the periphery and thymus of corrected autoimmune mice. AB - Fas (CD95) ligand (L) is a death factor that binds to its receptor, Fas, and induces apoptotic cell death, a crucial process in immunological tolerance. gld (generalized lymphoproliferative disorder) mice, which have a point mutation in the FasL gene, develop spontaneous systemic autoimmune syndromes characterized by hypergammaglobulinaemia and lymphoid hyperplasia owing to accumulation of abnormal B220+ CD3+ cells. Transplantation of wild-type (wt) bone marrow cells into old gld mice on the same strain background results in normalization of autoimmune syndromes. We characterized the cellular mechanisms (functionally and histologically) of the above phenomena in gld mice after bone marrow transplantation (BMT) to determine the role of apoptosis via Fas/FasL interactions in inducing and maintaining self-tolerance in vivo. Activated splenocytes from wt and BMT (wt to gld) mice showed significant cytotoxic activity against Fas transfectant cells while those from BMT (gld to gld) mice did not. Cells in the thymus, spleen and lymph nodes of gld mice uniformly upregulated Fas expression and were sensitive to Fas-mediated apoptosis compared with those in wt mice. Cells sensitive to Fas-mediated apoptosis in gld mice resided not only among abnormal B220+ CD3+ cells but also among conventional lymphocytes. More importantly, histological analysis revealed that cells in the spleen, lymph nodes and thymus frequently underwent apoptosis with infiltration of FasL+ cells in BMT (wt to gld) mice compared with BMT (gld to gld) mice. Our results indicated that apoptosis via Fas/FasL interactions can directly eliminate pathogenic cells responsible for autoimmunity in the periphery and possibly in the thymus in vivo. PMID- 9415030 TI - The polymeric immunoglobulin receptor (secretory component) in a human intestinal epithelial cell line is up-regulated by interleukin-1. AB - Secretory component (SC or polymeric immunoglobulin receptor) on mucosal epithelial cells mediates transcytosis of polymeric immunoglobulin into external fluids and functions as a receptor for polymeric immunoglobulin. SC expression in a human colonic adenocarcinoma cell line, HT-29 has been reported to be up regulated by various cytokines, such as interferon-gamma, tumour necrosis factor alpha and interleukin-4 (IL-4). However, up-regulation of SC by IL-1 is controversial. In this study, we investigated the effect of human recombinant IL 1 alone on SC expression in HT-29 cells in detail. Immunocytochemistry and Northern blot analysis revealed that IL-1 beta increased both the number of SC positive cells and SC mRNA expression. Enzyme-linked immunosorbent assay revealed that IL-1 beta enhanced secretion by HT-29 cells in both time- and dose-dependent manners. IL-1 alpha had the same effects on HT-29 cells. Northern blot analysis demonstrated that cycloheximide and actinomycin D abolished the effect of IL-1. Moreover, we detected IL-1 receptor (IL-1R) type I mRNA in HT-29 cells by polymerase chain reaction (PCR) and sequenced the PCR-amplified product. We think that it reflects the possibility of the presence of IL-1R in HT-29 cells. From these data, we concluded that IL-1 beta and IL-1 alpha play regulatory roles in SC expression, and their effects depend on de novo protein synthesis and transcription. PMID- 9415029 TI - L-selectin activates JNK via src-like tyrosine kinases and the small G-protein Rac. AB - Selectin and alpha 4 beta 7-integrins have been shown to mediate transient leucocyte interactions with endothelial cells which is a crucial step in the initial immune response to pathogens. We have previously shown that stimulation of T lymphocytes via L-selectin results in activation of a signalling cascade from the L-selectin molecule via the tyrosine kinase p56lck and tyrosine phosphorylation of L-selectin to the stimulation of p21Ras and Rac proteins. In the present study we demonstrate that stimulation of Jurkat T lymphocytes via L selectin results in an activation of Jun N-terminal kinase (JNK) but not of p38 K. L-selectin-initiated activation of JNK is mediated by src-like tyrosine kinases and the small G-protein Rac 1/2, since genetic or pharmacological inhibition of p56lck or Rac proteins prevent the stimulation of JNK by L selectin. Thus, the data point to a novel signalling cascade from L-selectin via src-like tyrosine kinases and Rac proteins to JNK. PMID- 9415031 TI - CD5+ B lymphocytes are the main source of antibodies reactive with non-parasite antigens in Trypanosoma congolense-infected cattle. AB - Mice infected with African trypanosomes produce exceptionally large amounts of serum IgM, a major part of which binds to non-trypanosome antigens such as trinitrophenol and single-strand DNA. In this paper, we describe that in cattle infected with Trypanosoma congolense and T. vivax, similar antibodies are found, although they bind mainly to protein antigens, such as beta-galactosidase, ovalbumin and ferritin. The parasite non-specific IgM antibodies appear around the same time as the parasite-specific antibodies, but their origin and function are not clear. We tested the hypothesis that CD5+ B cells (or B-1 cells), which increase during trypanosome infections in cattle, are responsible for production of antibodies to non-trypanosome antigens. Splenic CD5+ and CD5- B cells from infected cattle were sorted and tested in a single cell blot assay. The numbers of immunoglobulin-secreting cells were similar in both B-cell populations. However, antibodies with reactivity for non-trypanosome antigens were significantly more prevalent in the CD5+ B-cell fraction and were exclusively IgM. The preference for production of these antibodies by CD5+ B cells and the expansion of this subpopulation during infections in cattle, strongly suggest that CD5+ B cells are the main source of trypanosome non-specific antibodies. We propose that these antibodies are natural, polyreactive antibodies that are predominantly secreted by CD5+ B cells. Since B-1 cells are up-regulated in many states of immune insufficiency, the immunosuppression associated with trypanosome infections may be responsible for the increase of this subset and the concomitant increase in trypanosome non-specific antibodies. PMID- 9415032 TI - n-butyrate downregulates the stimulatory function of peripheral blood-derived antigen-presenting cells: a potential mechanism for modulating T-cell responses by short-chain fatty acids. AB - Modulation of proliferative T-cell responses by n-butyrate has been suggested to result from direct interference with cell cycle progression. Considering the important role of antigen-presenting cells (APC) in T-cell activation, we were particularly interested in studying the impact of n-butyrate on these cells. We demonstrated that pretreatment of human peripheral blood mononuclear cells (PBMC) or monocytes with this agent resulted in a dose- and time-dependent downregulation of their capability to stimulate T-cell responses with a similar pattern of inhibition when this agent was present throughout the culture period. Pretreatment with n-butyrate was effective in preventing both alloresponses and T cell proliferation to immobilized anti-CD3 monoclonal antibody (mAb) suggesting alteration of costimulatory function. Flow cytometric analysis revealed that interferon-gamma (IFN-gamma)-induced upregulation of B7-1 expression on monocytes was profoundly inhibited by n-butyrate. Furthermore, this agent significantly suppressed the expression of intercellular adhesion molecule-1 (ICAM-1) or lymphocyte function-associated antigen-3 (LFA-3). In contrast, constitutive as well as cytokine-induced expression of B7-2 was enhanced by n-butyrate. Additionally, in monocytes, but not in T cells, treatment with n-butyrate led to significant alteration of membrane integrity owing to apoptotic cell death. Our findings indicate that modulation of T-cell responses by n-butyrate could also result from altered APC function, possibly as a consequence of downregulating distinct adhesion and/or costimulatory receptors as well as of inducing apoptosis. A potential clinical relevance of short-chain fatty acids for reducing T-cell-mediated immune reactions via modulating APC function is speculated. PMID- 9415033 TI - Adhesion molecules are upregulated on dendritic cells isolated from human blood. AB - This study investigated whether the high expression of adhesion molecules on enriched preparations of circulating dendritic cells (DCs) was an intrinsic property of the cells or whether it was a consequence of the procedure used to isolate them from blood. Expression of the beta 1, beta 2 integrins (CD11/CD18 family) and other adhesion molecules on DCs in whole blood was compared with that on isolated DCs. Dendritic cells were identified by flow cytometry as leucocytes that were positive for human leucocyte antigen (HLA)-DR, but negative for CD3, CD14, CD16, CD19 and CD56. In contrast to a minority of DCs in whole blood, the majority of isolated DCs expressed the beta 2 integrins and there were a greater number of cells bearing CD44, CD54 and some of the beta 1 integrins (notably CD49b, CD49d, CD49e and CD29). An increase in the proportion of DCs bearing adhesion molecules was generally apparent at the isolation stage when mononuclear cells, which had been incubated overnight, were centrifuged on a metrizamide gradient to enrich for cells of low density. Inclusion of an inhibitor of protein glycosylation and exocytosis (brefeldin A) at all stages of separation partially prevented an increase in the percentage of DCs bearing CD18, C29 and C54 whereas the inclusion of cycloheximide (an inhibitor of polypeptide synthesis) interfered with increases in the percentage of cells bearing CD29 and CD54. Neither of these antagonists had an effect on the intensity of adhesion molecule expression. We suggest that some of the adhesion-dependent functions of isolated DCs are caused, in part, by an upregulation of surface adhesion molecules induced by the enrichment procedure. PMID- 9415034 TI - Role of spleen macrophages in innate and acquired immune responses against mouse hepatitis virus strain A59. AB - Owing to their scavenging and phagocytic functions, spleen macrophages are regarded to be important in the induction and maintenance of both innate and acquired immune defence mechanisms. In this study, we investigated the role of spleen macrophages in immunity against mouse hepatitis virus strain A59 (MHV A59). Previous studies showed that spleen and liver macrophages are the first target cells for infection by MHV-A59 in vivo, suggesting that they could be involved in the induction of immune responses against MHV-A59. We used a macrophage depletion technique to deplete macrophages in vivo and studied the induction of virus-specific antibody and cytotoxic T-cell (CTL) responses and non immune resistance against MHV-A59 in normal and macrophage-depleted mice. Virus titres in spleen and liver increased rapidly in macrophage-depleted mice, resulting in death of mice within 4 days after infection. Elimination of macrophages before immunization with MHV-A59 resulted in increased virus-specific humoral and T-cell proliferative responses. However, virus-specific CTL responses were not altered in macrophage-depleted mice. Our results show that spleen macrophages are of major importance as scavenger cells during MHV-A59 infection and are involved in clearance of virus from the host. In addition, macrophages may be involved in the regulation of acquired immune responses. In the absence of macrophages, increased virus-specific T-cell and antibody responses are detectable, suggesting that macrophages suppress MHV-A59-specific T- and B-cell responses and that other cells serve as antigen-presenting cells. PMID- 9415035 TI - Involvement of the membrane form of tumour necrosis factor-alpha in lipopolysaccharide-induced priming of mouse peritoneal macrophages for enhanced nitric oxide response to lipopolysaccharide. AB - We studied the pathways of macrophage response to lipopolysaccharide (LPS). When mouse macrophages pre-exposed to LPS were restimulated with this agent, reduced tumour necrosis factor-alpha (TNF-alpha) responses (desensitization/endotoxin tolerance) were accompanied by increased (priming) nitric oxide (NO) responses. Priming was also inducible with recombinant interferon-beta (IFN-beta). The requirement of TNF-alpha biosynthesis in the LPS-induced priming was also suggested by the observation that both anti-TNF-alpha serum and pentoxifylline inhibited this effect. However, addition of mouse recombinant TNF-alpha (mrTNF alpha) did not enhance the priming induced by LPS or IFN-beta, and preincubation with mrTNF-alpha alone, or in association with other cytokines produced by macrophages (interleukin-1 beta, interleukin-6, or leukaemia inhibitory factor), did not induce a priming effect. We found however, that pentoxifylline, which blocked the priming, also decreased the level of membrane-bound TNF-alpha. Furthermore, exposure to compound BB-3103 (a metalloproteinase inhibitor that blocks the processing of membrane-bound TNF-alpha yielding to the secreted cytokine) enhanced the priming effect, the expression of membrane TNF-alpha and the specific binding of LPS. These observations suggest that the membrane form of TNF-alpha is involved in the interaction of LPS with a receptor required for LPS induced priming. PMID- 9415036 TI - Tosylphenylalanine chloromethyl ketone inhibits TNF-alpha mRNA synthesis in the presence of activated NF-kappa B in RAW 264.7 macrophages. AB - Serine proteinase inhibitors such as N-tosyl-L-phenylalanine chloromethyl ketone (TPCK) and N alpha-p-tosyl-L-lysine chloromethyl ketone (TLCK) were shown to inhibit production of tumour necrosis factor-alpha (TNF-alpha) in lipopolysaccharide (LPS)-activated RAW 264.7 macrophages. The proteinase inhibitors were also reported to inhibit activation of the transcription factor nuclear factor-kappa B (NF-kappa B) by blocking the signalling pathway for stimuli-induced phosphorylation of the inhibitory subunit (I kappa B alpha) and thus preventing its degradation. In RAW 264.7 cells TPCK and TLCK significantly suppressed LPS-induced increase in TNF-alpha mRNA, induction of nuclear kappa B binding activity and degradation of I kappa B alpha. TPCK and TLCK effectively blocked TNF-alpha mRNA synthesis even when they were added after LPS stimulation. In these cells, however, the inhibitory modes of the two inhibitors were found to be different: while addition of TLCK suppressed I kappa B alpha degradation and reduced NF-kappa B activity, a comparable decrease in the nuclear kappa B-binding activity or in I kappa B alpha degradation was not observed in cells treated with TPCK. Our results show that TPCK inhibits LPS-induced TNF-alpha mRNA synthesis in the presence of activated NF-kappa B and suggests that mechanisms other than NF kappa B activation are involved in the transcriptional regulation of the TNF alpha gene. PMID- 9415037 TI - Macrophages activated by Listeria monocytogenes induce organ-specific autoimmunity. AB - We have previously reported an experimental autoimmune model induced by the local infection of Listeria monocytogenes. The unilateral inoculation of virulent Listeria into a testis of a normal mouse induced a delayed-type hypersensitivity response against testicular antigen and caused autoimmune orchitis in the contralateral testis. The orchitis was transferred to naive mice by T cells from the intratesticularly infected mice. In this paper, we demonstrated that avirulent Listeria, which lacks the expression of listeriolysin O, failed to induce any anti-testicular responses or contralateral orchitis even when it was inoculated at a high dose into the testis. Furthermore, the intraperitoneal inoculation of virulent Listeria with testicular antigen induced the anti testicular responses and orchitis although intraperitoneal inoculation of testicular antigen with avirulent Listeria failed to induce them. The difference between virulent and avirulent Listeria in the induction of anti-testicular responses was supposed to be dependent on the difference in macrophage activation by the two bacterial strains because, first, the anti-testicular responses were elicited in normal mice when macrophages from virulent Listeria-infected mice were intraperitoneally transferred with testicular antigen although no viable bacteria were detected from the macrophages, and secondly, in contrast, the intraperitoneal co-inoculation of macrophages from avirulent Listeria-infected mice and testicular antigen failed to elicit any anti-testicular responses. Finally, we found that the virulent Listeria-induced macrophages expressed a higher level of CD80 (B7-1) and CD86 (B7-2) molecules than did the avirulent Listeria-induced macrophages and naive peritoneal macrophages. These results thus suggest that virulent Listeria activates macrophages to induce autoreactive T cells while avirulent Listeria does not. The up-regulation of B7 molecules by virulent Listeria infection is a candidate of the mechanism for the activation of autoreactive T cells. PMID- 9415038 TI - Detection of a novel 40,000 MW excretory Toxoplasma gondii antigen by murine Th1 clone which induces toxoplasmacidal activity when exposed to infected macrophages. AB - To analyse target molecules of the CD4+ T-cell response to toxoplasma infection, a panel of Toxoplasma gondii-specific murine CD4+ T-cell clones has been established. Clone 3Tx15, belonging to the T helper 1 (Th1) subtype, abolished intracellular parasite growth when co-cultured with macrophages and live toxoplasma at a ratio of 2:2:1. This effect results from macrophage toxoplasmicidal activity induced upon parasite-dependent cellular interaction, an irrelevant Th1 clone failed in this three-party system. Clone 3Tx15 detects its corresponding antigen in the supernatant of infected cells and also reacts with a host cell-free preparation of T. gondii-excreted/secreted antigens. T-cell blot analysis of two-dimensionally separated toxoplasma lysate revealed a molecular weight of about 40,000 for the fractions stimulating clone 3Tx15. As checked in parallel enzyme-linked immunosorbent assay, the 40,000 MW T-cell antigen co migrates with the excretory protein GRA4, the sole 40,000 MW T. gondii antigen hitherto known to be recognized by T lymphocytes. Nevertheless, neither recombinant GRA4 nor immunoaffinity-purified natural GRA4 was stimulatory for clone 3Tx15. Our findings thus demonstrate that Th1 clone 3Tx15 which induces toxoplasmicidal activity during antigenic interaction with infected macrophages defines a new 40,000 MW excretory T. gondii antigen. PMID- 9415039 TI - Role of E- and P-selectin in migration of monocytes and polymorphonuclear leucocytes to cytokine and chemoattractant-induced cutaneous inflammation in the rat. AB - The contribution of E- and P-selectin in the rat to the migration of polymorphonuclear leucocytes (PMNL) and monocytes to acute dermal inflammation induced by a chemoattractant (C5ades Arg) or endothelial cell activating agents [lipopolysaccharide, tumour necrosis factor-alpha (TNF-alpha), alpha-thrombin and interferon-gamma] administered intradermally was investigated. Migration was quantitated using radiolabelled blood PMNL and monocytes and new, function blocking monoclonal antibodies (mAbs) to rat E- and P-selectin were employed. Monocyte migration to inflamed skin was partially inhibited (40-75%) by P selectin mAb with all stimuli tested, but not by anti-E-selectin. PMNL migration in response to all stimuli was also inhibited (50-75%) by blocking P-selectin, but in contrast to monocytes, PMNL accumulation was partially inhibited by mAb to E-selectin in alpha-thrombin and TNF-alpha lesions. When P-selectin was blocked by mAb, mAb to E-selectin significantly inhibited further (by 70-90%) both PMNL and monocyte accumulation in all lesions, indicating that both P- and E-selectin contribute to the migration of these leucocytes. Blocking L-selectin in addition to P- and E-selectin, had no effect on the remaining migration. Thus, optimal PMNL and monocyte migration to chemotactic factor- and cytokine-induced skin inflammation is P-selectin dependent. E-selectin becomes important, in most conditions used here, when P-selectin mediated recruitment is not operative. A selectin independent pathway likely mediates up to 20% of PMNL and monocyte migration to acute inflammation, at least in skin. PMID- 9415042 TI - Assessment of human energy balance. PMID- 9415040 TI - Augmentation of monocyte chemotactic protein-1 and mRNA transcript in chronic inflammatory states induced by potassium permanganate (KMnO4) in vivo. AB - Monocyte chemotactic protein-1 (MCP-1) is a proinflammatory cytokine that attracts and activates specific types of leucocytes. The purpose of this work was to analyse the generation of MCP-1 and mRNA transcript in a model of chronic inflammation using a granulomatous tissue induced by potassium permanganate (KMnO4; water soluble crystals). The data presented here shows that MCP-1 is generated in granuloma tissue and its level was strongly increased by i.p. injections of lipopolysaccharide (LPS) and inhibited in rats treated with injections of dexamethasone, 18 hr before the animals were killed. In histological studies LPS and dexamethasone increased and decreased, respectively, the recruitment of mononuclear cells in the granuloma tissue compared with the control granulomas from phosphate-buffered saline (PBS)-treated animals. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used for mRNA extraction and cDNA synthesis. mRNA MCP-1 was significantly produced in the granuloma tissue of untreated animals, an effect increased by LPS and inhibited by dexamethasone, compared with the controls. Moreover, MCP-1 protein was found in the supernatant from homogenized granuloma tissues and the levels of MCP-1 were higher in the LPS treated animals, while they were lower in the dexamethasone group, compared with the granulomas from the PBS-treated groups (control). The generation of MCP-1 was also found in minced granuloma tissue incubated for 18 hr (overnight) from treated (LPS or dexamethasone) and untreated (PBS) rats. When LPS was added in vitro for 18 hr to the controls and treated animals the production of MCP-1 was further increased except in the dexamethasone group (P > 0.05). Analysing blood serum from LPS, dexamethasone or PBS-treated rats, we found that MCP-1 was also present. The level was higher in the LPS group and lower in the dexamethasone group, compared with the control (PBS). In these studies we show for the first time that MCP-1 transcript and translation is generated in chronic experimental inflammatory tissue, an effect inhibited by dexamethasone. PMID- 9415043 TI - Investigation of human adipose tissue metabolism in vivo. PMID- 9415044 TI - Regional adipocity in man. PMID- 9415041 TI - The species-specific structure of microanatomical compartments in the human spleen: strongly sialoadhesin-positive macrophages occur in the perifollicular zone, but not in the marginal zone. AB - The microanatomical structure of human and rat splenic white pulp is compared, with special emphasis on the localization of the marginal zone occupied by immunoglobulin M (IgM)+ IgD-/dull B lymphocytes and its specialized macrophages. Our study reveals that in contrast to rats, the marginal zone of humans primarily exists in the vicinity of primary and secondary splenic follicles and that it is almost absent around the periarteriolar T-cell zones. We demonstrate that in humans there is an additional compartment, the perifollicular zone, located between the marginal zone and the red pulp. The perifollicular zone is a dynamic region of variable cellular and phenotypic composition, which can be regarded either as a part of the red pulp or of the follicles. In most cases the perifollicular zone appears as a compartment of the red pulp containing erythrocyte-filled spaces which differ from the typical red pulp sinusoids. Similar to the splenic cords, the perifollicular zone mostly harbours scattered B and T lymphocytes. However, sometimes B lymphocytes clearly predominate in the perifollicular area. In addition, strongly sialoadhesin-positive macrophages form sheaths around capillaries in the perifollicular zone. Such capillary sheaths are not observed in rats. In humans weakly sialoadhesin-positive macrophages are also present in the perifollicular zone and in the red pulp. In some specimens sialoadhesin is, however, strongly expressed by a large number of dispersed perifollicular macrophages. Interestingly, in striking contrast to rats, the human marginal zone does not contain sialoadhesin-positive macrophages and marginal metallophilic macrophages are also absent in humans. Thus, sialoadhesin positive macrophages and IgM+ IgD- memory B lymphocytes both share the marginal zone as a common compartment in rats, while they occupy different compartments in humans. We show that the human splenic marginal zone does not contain a marginal sinus and assume that in humans the perifollicular region is the compartment where antigen and recirculating lymphocytes enter the organ. PMID- 9415045 TI - Neuroendocrine factors in obesity. PMID- 9415046 TI - Control of appetite--the role of glucagon-like peptide-1 (7-36) amide. PMID- 9415047 TI - Molecular mechanisms of adipocyte differentiation. PMID- 9415048 TI - Acylation stimulating protein and the adipocyte. AB - ASP constitutes a metabolic bridge integrating events in the circulation with the adipocyte microenvironment and regulation of a key adipose function-storage of fat. Dysregulation of the ASP pathway may have important metabolic consequences and may be associated with both obesity on one hand and cardiovascular disease on the other hand. Clearly, many in vitro and in vivo studies remain to be done to establish clearly such links and future work on ASP promises to be both exciting and rewarding. PMID- 9415049 TI - Role of the agouti gene in obesity. PMID- 9415050 TI - Carboxypeptidase E and obesity in the mouse. PMID- 9415051 TI - Leptin and its receptor. PMID- 9415052 TI - PPAR gamma and the molecular control of adipogenesis. PMID- 9415053 TI - Studies of the mechanism of inhibition of insulin signaling by tumor necrosis factor-alpha. PMID- 9415054 TI - Towards the development and use of human-selective agonists for the pharmacologic treatment of obesity and diabetes. PMID- 9415055 TI - Obesity and diabetes--potential for intervention? PMID- 9415056 TI - Regulation of IGF-binding protein-6 by dexamethasone and IGFs in PC12 rat phaeochromocytoma cells. AB - PC12 rat phaeochromocytoma cells are widely used as a model of neuronal differentiation. They express IGF receptors and are responsive to IGFs. The main IGF-binding protein synthesized by these cells is IGFBP-6. Glucocorticoids induce differentiation of PC12 cells towards a chromaffin phenotype. The effect of dexamethasone on IGFBP-6 levels was therefore studied. Dexamethasone (500 nM) decreased IGFBP-6 protein in conditioned media and mRNA levels to 61 +/- 5% (P < 0.0001) and 34 +/- 14% (P = 0.03) of control levels respectively. Incubation of PC12 cells with IGF-II (100 ng/ml) for 72 h increased IGFBP-6 protein levels in media to 217 +/- 19% of control (P < 0.0001). IGFBP-6 mRNA levels, however, were unchanged. IGF-I had similar effects on IGFBP-6 protein and mRNA levels. IGFs increased cell number by 50-60%, but this was insufficient to explain the increases in protein levels. IGFBP-6 was not released from a cell-associated reservoir or protected from proteolysis by IGFs, excluding these post translational mechanisms as explanations for the IGF effects on IGFBP-6 levels. The effects of IGF-II and dexamethasone on IGFBP-6 levels were independent. These results indicated that (1) dexamethasone decrease IGFBP-6 at the mRNA level, and (2) IGFs stimulate IGFBP-6 levels by a post-transcriptional mechanism. PMID- 9415057 TI - Dehydroepiandrosterone administration prevents the oxidative damage induced by acute hyperglycemia in rats. AB - Free radical overproduction contributes to tissue damage induced by acute hyperglycemia. Dehydroepiandrosterone, which has recently been found to have antioxidant properties, was administered i.p. to rats at different doses (10, 50 or 100 mg/kg body weight) 3 h before treatment with dextrose (5 g/kg). Lipid peroxidation was evaluated on liver, brain and kidney homogenates, measuring both steady-state concentrations of thiobarbituric acid reactive substances, and fluorescent chromolipids, evaluated as hydroxynonenal adducts. Formation of thiobarbituric acid reactive substances was significantly higher in hyperglycemic than in normoglycemic animals. Three hours (but not 1 h) dehydroepiandrosterone pretreatment protected tissues against lipid peroxidation induced by dextrose; both thiobarbituric acid reactive substances and hydroxynonenal adducts in liver, kidney and brain homogenates were significantly lower in dehydroepiandrosterone pretreated animals. Dehydroepiandrosterone did not modify the cytosolic level of antioxidants, such as alpha-tocopherol or glutathione, nor the activities of glutathione peroxidase, reductase or transferase. The results of this study indicate that the 'in vivo' administration of dehydroepiandrosterone increases tissue resistance to lipid peroxidation triggered by acute hyperglycemia. PMID- 9415058 TI - Evidence that the alpha-subunit influences the specificity of receptor binding of the equine gonadotrophins. AB - Horse LH/chorionic gonadotrophin (eLH/CG) exhibits, in addition to its normal LH activity, a high FSH activity in all other species tested. Donkey LH/CG (dkLH/CG) also exhibits FSH activity in other species, but about ten times less than the horse hormone. In order to understand the molecular basis of these dual gonadotrophic activities of eLH/CG and dkLH/CG better, we expressed, in COS-7 cells, hybrids between horse and donkey subunits, between horse or donkey alpha subunit and human CG beta (hCG beta), and also between the porcine alpha-subunit and horse or donkey LH/CG beta. The resultant recombinant hybrid hormones were measured using specific FSH and LH in vitro bioassays which give an accurate measure of receptor binding specificity and activation. Results showed that it is the beta-subunit that determines the level of FSH activity, in agreement with the belief that it is the beta-subunit which determines the specificity of action of the gonadotrophins. However, donkey LH/CG beta combined with a porcine alpha subunit exhibited no FSH activity although it showed full LH activity. Moreover, the hybrid between horse or donkey alpha-subunit and hCG beta also exhibited only LH activity. Thus, the low FSH activity of dkLH/CG requires an equine (donkey or horse) alpha-subunit combined with dkLH/CG beta. These results provide the first evidence that an alpha-subunit can influence the specificity of action of a gonadotrophic hormone. PMID- 9415059 TI - The effects of pregnancy steroids on adaptation of beta cells to pregnancy involve the pancreatic glucose sensor glucokinase. AB - Pregnancy is associated with adaptive changes including increased number and size of beta cells and enhanced gap-junctional coupling among beta cells, increased glucose-induced insulin response and decreased glucose stimulation threshold. The role exerted by pregnancy steroids and lactogenic hormones in the development of islets upregulation during pregnancy has been widely investigated. In the present study we studied the possibility that pregnancy steroids induce functional modifications of beta cells involving the expression and function of glucokinase. Our results indicate that estradiol and progesterone do not influence significantly glucokinase mRNA expression, while they induce a dose-dependent and time-dependent increase of glucokinase activity in RIN 1046-38 cells. The increased enzymatic activity results in an increased glucose-induced insulin release. Therefore it is possible to hypothesize that pregnancy steroids influence glucokinase expression in beta cells at a post-transcriptional level and that this effect contributes to the development of hyperinsulinemia during pregnancy. PMID- 9415061 TI - Distribution of prolactin receptor immunoreactivity in ovine skin and changes during the wool follicle growth cycle. AB - Prolactin is believed to mediate seasonal cues entraining seasonal reproductive and hair follicle growth cycles. Prolactin receptor binding activity and prolactin receptor gene expression in mammalian skin have recently been described. In this report, prolactin receptor immunoreactivity is identified in sheep skin using a monoclonal antibody against the rat liver prolactin receptor. Western blotting analysis of microsomal membrane proteins from skin showed major bands corresponding to molecular weights of 87 and 71 kDa and minor bands at 101 and 21 kDa. RNase protection analysis revealed the presence of mRNA species coding for long and short forms of the prolactin receptor. Formalin-fixed sections, exposed to the monoclonal antibody and stained by an immunogold method, revealed prolactin receptor-immunoreactivity in the dermal papilla, germinal matrix, outer root sheath, lower regions of the inner root sheath and connective tissue sheath of wool follicles. Staining was absent from keratinised cell populations. In all samples, the interfollicular epidermis, sebaceous and sweat glands were positively stained. The distribution of prolactin receptor is described in both growing and inactive wool follicles and related to postulated cycle-specific actions of circulating prolactin in the control of seasonal fibre growth. PMID- 9415060 TI - Selenium and iodine deficiencies: effects on brain and brown adipose tissue selenoenzyme activity and expression. AB - Adequate dietary iodine supplies and thyroid hormones are needed for the development of the central nervous system (CNS) and brown adipose tissue (BAT) function. Decreases in plasma thyroxine (T4) concentrations may increase the requirement for the selenoenzymes types I and II iodothyronine deiodinase (ID-I and ID-II) in the brain and ID-II in BAT to protect against any fall in intracellular 3,3',5 tri-iodothyronine (T3) concentrations in these organs. We have therefore investigated selenoenzyme activity and expression and some developmental markers in brain and BAT of second generation selenium- and iodine deficient rats. Despite substantial alterations in plasma thyroid hormone concentrations and thyroidal and hepatic selenoprotein expression in selenium and iodine deficiencies, ID-I, cytosolic glutathione peroxidase (cGSHPx) and phospholipid hydroperoxide glutathione peroxidase (phGSHPx) activities and expression remained relatively constant in most brain regions studied. Additionally, brain and pituitary ID-II activities were increased in iodine deficiency regardless of selenium status. This can help maintain tissue T3 concentrations in hypothyroidism. Consistent with this, no significant effects of iodine or selenium deficiency on the development of the brain were observed, as assessed by the activities of marker enzymes. In contrast, BAT from selenium- and iodine deficient rats had impaired thyroid hormone metabolism and less uncoupling protein than in tissue from selenium- and iodine-supplemented animals. Thus, the effects of selenium and iodine deficiency on the brain are limited due to the activation of the compensatory mechanisms but these mechanisms are less effective in BAT. PMID- 9415062 TI - Postnatal decline in gonadal secretion of dehydroepiandrosterone and 3 beta hydroxyandrosta-5,7-dien-17-one in the newborn foal. AB - Dehydroepiandrosterone (DHEA) and 3 beta-hydroxyandrosta-5,7-dien-17-one (7 dehydro-DHEA) are secreted in large quantities by the remarkably hypertrophied fetal gonads of both sexes in the pregnant mare. Their secretion serves as the fetal component of a feto-placental unit for oestrogen production in equine pregnancies. They are secreted in large amounts but show a decline in late pregnancy when the fetal gonads regress and levels of oestrogens in the mare fall as a consequence. We have examined the levels of these precursor steroids in the newborn foal in the first days after birth. DHEA and 7-dehydro-DHEA were measured in peripheral plasma in a direct RIA with a DHEA antibody which cross-reacts with 7-dehydro DHEA (> 150%). Subsequent studies were performed with solid-phase extraction, separation of unconjugated from conjugated steroids, and HPLC fractionation followed by RIA. Detection on HPLC at 254 and 280 nm was compared with results from RIA. It was concluded that DHEA is the major steroid produced by the gonads at birth. The concentrations are highly variable in the first day of postnatal life (70.45 +/- 63.06 ng/ml, n = 52) and decline rapidly to < 2 ng/ml (n = 6) at 96 h after birth. At this time the sulphate form is also seen, with an increasing ratio of DHEAS/DHEA as the value for total DHEA falls. The mechanism and significance of the apparent abrupt decline in gonadal steroidogenesis in the newborn foal remain unknown. PMID- 9415063 TI - Comparative toxicity of alloxan, N-alkylalloxans and ninhydrin to isolated pancreatic islets in vitro. AB - The in vitro toxicity of the diabetogenic agent alloxan as documented by the induction of beta cell necrosis was studied in isolated ob/ob mouse pancreatic islets. The effect of alloxan has been compared with that of a number of N-alkyl alloxan derivatives and with that of the structurally related compound, ninhydrin. Alloxan and its derivatives were selectively toxic to pancreatic beta cells, with other endocrine cells and exocrine parenchymal cells being well preserved, even at high concentration. In contrast, ninhydrin was selectively toxic to pancreatic beta cells only at comparatively low concentration, destroying all islet cell types at high concentrations. The ultrastructural changes induced by all the test compounds in pancreatic beta cells in vitro were very similar to those observed during the development of alloxan diabetes in vivo. The relative toxicity of the various compounds to pancreatic beta cells in vitro was not, however, related to their ability to cause diabetes in vivo. Indeed, the non-diabetogenic substances ninhydrin, N-butylalloxan and N isobutylalloxan were very much more toxic to isolated islets than the diabetogenic compounds alloxan and N-methylalloxan. These results suggest that the differences in diabetogenicity among alloxan derivatives are not due to intrinsic differences in the susceptibility of the pancreatic beta cells to their toxicity, but may reflect differences in distribution or metabolism. High concentrations of glucose protected islets against the harmful effects of alloxan and its derivatives, but not those of ninhydrin. Low levels of glucose, and non carbohydrate nutrients, afforded little protection, indicating that the effect of glucose is not due to the production of reducing equivalents within the cell, 3-O Methylglucose, which protects against alloan diabetes in vivo, did not protect against alloxan toxicity in vitro. Since 3-O-methylglucose is known to prevent uptake of alloxan by pancreatic beta cells, it appears that uptake of alloxan by the cell is not a prerequisite for the induction of beta cell necrosis. PMID- 9415064 TI - Divergent deiodination of thyroid hormones in the separated parts of the fetal and maternal placenta in pigs. AB - Previous work from this laboratory has shown that the thyroid gland of the fetal pig begins to function at about day 46-47 (0.40-0.415 fraction of gestational age). Sera from fetuses contain lower thyroxine (T4), 3,3',5-triiodothyronine (T3) and 3,3',5'-triiodothyronine (rT3) concentrations than maternal sera, except for about 2 weeks before term. The fetal T4 metabolism is dominated by the 5' monodeiodinating activity (5'-MD). In the present study we measured the iodothyronines content, and the outer (5'-MD) and inner (5-MD) monodeiodinases activity, in homogenates of the placenta. The pig placenta, which is of the epitheliochorial type, was separated into the fetal and the maternal part. The concentrations of T4, T3 and rT3 were lower, and the deiodinating activity of 5' MD and 5-MD higher, in the fetal than in the maternal placenta. The fetal placenta not only deiodinated more actively T4 to T3 and T4 to rT3, but degraded T3 to 3,3'-diiodothyronine (3,3'-T2) more actively than rT3 to 3,3'-T2. Such divergent deiodinating activity of T4 to T3, T3 to 3,3'-T2 and rT3 to 3,3'-T2 might favor establishing a relatively high and constant rT3 concentrations in fetal and maternal placentas, and a lower T3 in the fetal placenta. The inner ring deiodinating activity (excluding a day before parturition) was always more active in the fetal placenta, while the outer ring deiodinations varied in this respect, depending on the gestation stage. These results support the hypothesis that in the fetal pig, enzymatic deiodination of thyroid hormones forms a barrier which reduces transplacental passage of the hormones and that the fetal part of the placenta is the primary factor in the mechanism regulating the hormonal transfer. In spite of the presence of the barrier, there is an adequate maternal supply of thyroid hormones to the fetus in early gestation, which suggests that the enzymatic mechanism is influenced in some way by the thyroid status of the fetus. PMID- 9415065 TI - Differential adrenergic regulation of rat pineal cyclic AMP production and N acetyltransferase activity during postnatal development: involvement of G alpha s and G alpha i1-2 proteins. AB - We have studied why rat pineal N-acetyltransferase (NAT) activity is relatively insensitive to isoproterenol in young rats when compared with adult rats. We report that activation by isoproterenol of pineal cyclic AMP production and NAT activity is higher in adult than in 2-week-old rats. However, the effect of dibutyryl cyclic AMP, which enters the pinealocyte and duplicates the effect of cyclic AMP, on NAT activity was similar at both ages. Moreover, we found that both alpha- and beta-adrenergic receptors are highly specific at both ages, since the binding of the specific radioligands used to their receptors could be displaced only by their corresponding agonists and antagonists. However, we observed differences between pineals from young and adult rats when several families of the alpha subunit of G-proteins were studied in cell membranes. ADP ribosylation and immunoblot studies have shown clear differences in both 42 and 45 kDa forms of the Gs alpha Both forms exhibit low values in pineals from 2-week old animals when compared with 6-week-old. We also show that the later appearance of both Gs alpha forms is roughly similar to the potent activation of cyclic AMP production and NAT activity in adult rats when compared with young rats. In conclusion, the results presented suggest that the relative lack of sensitivity of rat pineal gland to beta-adrenergic receptor agonists early in the postnatal development may be explained by the low levels of membrane Gs alpha, rather than postreceptor-mediated mechanisms or changes in the characteristics of the beta adrenergic receptors on the pinealocyte membrane. PMID- 9415066 TI - Cellular distribution of insulin-like growth factor binding protein mRNAs and peptides during rat lung development. AB - A role for IGF binding proteins (IGFBPs) in lung development is suggested by the identification of IGFBPs in lung tissue and production of IGFBPs by fetal lung cells in culture. To characterize the expression of IGFBPs during lung development in the rat in vivo (16 days gestation through adulthood), the expression of IGFBP mRNAs (IGFBP-1 to IGFBP-6) was examined by Northern analysis and in situ hybridization, and IGFBP peptides (IGFBP-2, IGFBP-3, and IGFBP-5) were localized by immunohistochemistry. IGFBP-1 mRNA was not detectable. IGFBP-2 mRNA (1.8 kb) was expressed in both fetal and postnatal life with peak expression during the fetal pseudoglandular stage. IGFBP-2 mRNA was localized mainly to airway epithelium. IGFBP-3 mRNA (2.4 kb) was maximally expressed postnatally in the saccular stage of lung development; it was identified in airway epithelium and interstitium in the fetal lung, but predominantly in airway epithelium after birth. IGFBP-4 (2.6 kb) and IGFBP-5 (6.0 kb) mRNA levels were maximal after birth, from 3 to 21 days postnatal (saccular and alveolar stage). IGFBP-4 mRNA was localized primarily to the interstitium and blood vessels early in development, but was abundant in airway epithelium in the adult. IGFBP-5 mRNA was most abundant in the airway epithelium. IGFBP-3, IGFBP-4, IGFBP-5, and to a lesser extent IGFBP-6 were localized to the large cartilaginous airways in the adult. IGFBP-2, IGFBP-3, and IGFBP-5 peptides were distributed more widely than their respective mRNAs, with a temporal pattern of immunoreactivity following that of their mRNAs. Maximal staining was noted in airway epithelium for IGFBP-2 in the newborn, for IGFBP-3 in the saccular stage (newborn to 3 days postnatal), and for IGFBP-5 in the alveolar stage (5 to 21 days postnatal). Our studies demonstrate that IGFBP-2, IGFBP-3, IGFBP-4, and IGFBP-5 are synthesized and distributed in spatially and temporally different patterns in the developing lung. The widespread distribution of IGFBP immunoreactivity compared with their respective mRNAs suggests that IGFBPs are important paracrine factors in the regulation of IGF action in the developing lung. PMID- 9415067 TI - The expression of regulated endocrine-specific protein of 18 kDa in peptidergic cells of rat peripheral endocrine tissues and in blood. AB - We examined the cellular localization of regulated endocrine-specific protein of 18 kDa (RESP18) and mRNA in peripheral endocrine tissues. In situ hybridization and immunocytochemistry identified RESP18 mRNA in most cells of the anterior and intermediate pituitary, with RESP18 protein apparent in many anterior pituitary cells but very few intermediate pituitary cells. In the adrenal medulla and superior cervical ganglion, RESP18 mRNA co-localized with dopamine beta-mono oxygenase and neuropeptide Y. In the thyroid, RESP18 mRNA was localized to C cells. RESP18 mRNA was expressed in most of the cells of the pancreatic islets, co-localizing with insulin, glucagon, and somatostatin. No RESP18 mRNA or protein was detected in the adrenal cortex, ovary, neural lobe of the pituitary, parathyroid, exocrine pancreas, thyroid follicular cells, placenta, mammary tissue, liver, lung, or atria. As in the intermediate lobe of the pituitary, high levels of RESP18 mRNA in the pancreatic islets and adrenal medulla did not always correlate with immunodetectable RESP protein, suggesting that post transcriptional mechanisms are important in controlling RESP18 expression. Western blot analyses identified 18 kDa RESP and higher molecular weight isoforms of RESP in most tissues and in plasma. Subcellular fractionation of the anterior pituitary identified 18 kDa RESP18 in fractions enriched in endoplasmic reticulum and secretory granules, with the higher molecular weight isoforms of RESP18 concentrated in fractions enriched in secretory granules. The broad neuroendocrine distribution of RESP18 suggests that it subserves an important function in the secretory pathway that is common to the production of many secreted peptides. PMID- 9415068 TI - Effects of cytochrome P450 inhibitors and of steroid hormones on the formation of 7-hydroxylated metabolites of pregnenolone in mouse brain microsomes. AB - Hydroxylations of pregnenolone (PREG) at the 7 alpha- and 7 beta-positions have been reported in numerous murine tissues and organs and responsible cytochrome P450 (CYP) species await identification. Using thin layer chromatography and gas chromatography-mass spectrometry, we report identification of 7 alpha-hydroxy PREG and 7 beta-hydroxy-PREG metabolites produced in mouse brain microsome digests and kinetic studies of their production with apparent KM values of 0.5 +/ 0.1 microM and 5.1 +/- 0.6 microM for 7 alpha- and 7 beta-hydroxylation respectively. Investigation of CYP inhibitors and of steroid hormone effects on both 7 alpha- and 7 beta-hydroxylations of PREG showed that: (i) different CYP were involved in 7 alpha- and 7 beta-hydroxylation of PREG because solely 7 alpha hydroxylation was extensively inhibited by metyrapone, alpha-naphthoflavone, ketoconazole and 3 beta-hydroxysteroids, (ii) CYP 1A2, 2D6, 2B1 and 2B11 were not responsible for 7 alpha- and 7 beta-hydroxylation of PREG because respective specific inhibitors furafylline, quinidine and chloramphenicol triggered no inhibition, (iii) CYP 1A1 was responsible for only part of the 7 beta hydroxylation of PREG because use of alpha-naphthoflavone, which inhibits specifically CYP 1A1, did not suppress entirely 7 beta-hydroxylation, while ketoconazole, metyrapone and antipyrine, which do not inhibit CYP 1A1, decreased part of the 7 beta-hydroxylation, (iv) 7 alpha-hydroxylation of PREG may be shared with other 3 beta-hydroxysteroids such as isoandrosterone and 5-androstene 3 beta,17 beta-diol which were strong inhibitors, but not with dehydroepiandrosterone which was a non-competitive inhibitor as weak as 3 oxosteroids, and (v) 7 beta-hydroxylation of PREG was not markedly changed by other steroids. Taken together, these findings will be of use for identification of the CYP species responsible for 7 alpha- and 7 beta-hydroxylation of PREG and for studies of their activities in brain. PMID- 9415069 TI - Effects of insulin-like growth factor-I and LR3IGF-I on regional blood flow in normal rats. AB - Two studies were conducted to investigate the haemodynamic effects of IGF-I and its analogue LR3IGF-I in normal anaesthetised rats. Infusion of IGF-I intravenously, at a dose of 125 micrograms/kg/h, for 20 min in the first study resulted in renal blood flow being significantly elevated by 35% above baseline. Mean arterial blood pressure (MABP) at this IGF-I dose fell by 18% of baseline, with LR3IGF-I also causing a significant decline in MABP (by 15%) at the dose of 125 micrograms/kg/h. In the second study the intravenous administration of IGF-I or LR3IGF-I, at a dose of 125 micrograms/kg/h, over a period of 60 min, resulted in MABP being significantly lowered by 25% of baseline values. Regional blood flow rates were determined using radioactive microspheres, 15 microns in diameter, injected systemically at the end of the peptide infusion period. The gastrocnemius, a representative skeletal muscle, was the only vascular region to show a significant increase in blood flow after IGF-I (by 58%) or LR3IGF-1 (by 308%) infusion. Vascular resistance in the brain was significantly reduced after infusion of IGF-I (by 60%) or LR3IGF-I (by 48%) as compared with vehicle. Skeletal muscle vascular resistance was also reduced by IGF-I (by 41%) and more particularly by LR3IGF-I (by 77%) in comparison to vehicle. These alterations to vascular tone produced by IGF infusion may be related to the central nervous system and systemic cardiovascular side-effects that have been reported during IGF-I administration in humans. PMID- 9415070 TI - Maternal endocrine status in relation to pregnancy outcome in rapidly growing adolescent sheep. AB - It has previously been reported that high nutrient intakes which promote rapid maternal growth throughout pregnancy are associated with poor pregnancy outcome when compared with normally growing adolescent animals. The present study examined the maternal plasma concentrations of a number of putative endocrine regulators of nutrient partitioning between the maternal and fetal compartments in relation to placental and fetal growth in this novel experimental paradigm. Embryos were recovered on day 4 after oestrus from superovulated adult ewes that had been inseminated using semen from a single sire and synchronously transferred, in singleton, to the uterus of peripubertal adolescent recipients (n = 38), which had been induced to ovulate at 32 weeks of age (live weight 47.4 +/- 0.4 kg). Post-transfer, the adolescent recipients were offered a high (n = 21) or moderate (n = 17) level of a complete diet calculated to achieve rapid (RMG) or normal (NMG) maternal growth rates. After day 100 of gestation, the feed intake of the NMG group was adjusted weekly to meet the increasing nutrient demands of the gravid uterus. Pregnancy rate following embryo transfer was higher (P < 0.05) in the RMG (90%) than in the NMG (59%) group. For ewes delivering live young at term, liveweight gain during the first 100 days of gestation was 294 +/- 12.9 and 84 +/- 4.7 g/day for the RMG (n = 16) and NMG (n = 10) groups respectively, and body condition score immediately prior to parturition was higher in RMG than in NMG ewes (2.9 +/- 0.04 vs 1.9 +/- 0.15 score units respectively, P < 0.001). For the RMG and NMG groups respectively, mean placental weight was 327 +/- 18.1 and 485 +/- 16.6 g with lamb birth weights of 3.49 +/- 0.13 and 4.82 +/- 0.21 kg (P < 0.001). The reduction in placental mass in the RMG group reflected a decrease in the number (P < 0.001) and size (P < 0.01) of the fetal cotyledons. The duration of gestation was shorter (P < 0.001) and colostrum yield at parturition lower (P < 0.001) in the RMG group. Maternal insulin concentrations, determined three times weekly, were higher (P < 0.001) throughout gestation in the RMG group and irrespective of treatment group were negatively correlated (P < 0.01) with placental weight and lamb birth weight. High glucose levels throughout gestation and a decreased response to an exogenous insulin challenge on day 95 of gestation implied a degree of insulin resistance in the RMG group but, in spite of these high maternal glucose concentrations, the reduced size of the placenta probably constrained fetal growth. Maternal IGF-I levels determined weekly, were elevated (P < 0.001) during the second and third trimester in RMG versus NMG groups and a sustained elevation in maternal tri-iodothyronine and thyroxine concentrations was evident in the RMG group from mid-gestation. In contrast, GH pulse frequency and mean GH concentrations, determined on day 68 and 122 of gestation, were lower (P < 0.05) in the RMG group, and irrespective of treatment group, were correlated negatively with feed intake and positively with placental weight and colostrum yield at parturition. Progesterone concentrations were lower in the RMG group during the second and third trimesters (P < 0.001) and, irrespective of treatment group, were positively associated (P < 0.001) with placental weight, gestation length and colostrum yield. These results suggest that in pregnant adolescent sheep on high dietary intakes, elevated insulin and IGF-I levels ensure that the anabolic drive to maternal tissue synthesis is established during early gestation at the expense of placental growth. The consequent restriction in placental transport capacity is the primary limitation to fetal growth and reduced GH and placental progesterone secretion may impair colostrum production. PMID- 9415071 TI - Glucagon-like peptide-1(7-36)-amide confers glucose sensitivity to previously glucose-incompetent beta-cells in diabetic rats: in vivo and in vitro studies. AB - The effects of glucagon-like peptide-1(7-36)-amide (GLP-1) on cAMP content and insulin release were studied in islets isolated from diabetic rats (n0-STZ model) which exhibited impaired glucose-induced insulin release. We first examined the possibility of re-activating the insulin response to glucose in the beta-cells of the diabetic rats using GLP-1 in vitro. In static incubation experiments, GLP-1 amplified cAMP accumulation (by 170%) and glucose-induced insulin release (by 140%) in the diabetic islets to the same extent as in control islets. Using a perifusion procedure, GLP-1 amplified the insulin response to 16.7 mM glucose by diabetic islets and generated a clear biphasic pattern of insulin release. The incremental insulin response to glucose in the presence of GLP-1, although lower than corresponding control values (1.56 +/- 0.37 and 4.53 +/- 0.60 pg/min per ng islet DNA in diabetic and control islets respectively), became similar to that of control islets exposed to 16.7 mM glucose alone (1.09 +/- 0.15 pg/min per ng islet DNA). Since in vitro GLP-1 was found to exert positive effects on the glucose competence of the residual beta-cells in the n0-STZ model. we investigated the therapeutic effect of in vivo GLP-1 administration on glucose tolerance and glucose-induced insulin release by n0-STZ rats. An infusion of GLP 1 (10 ng/min per kg; i.v.) in n0-STZ rats enhanced significantly (P < 0.01) basal plasma insulin levels, and, when combined with an i.v. glucose tolerance and insulin secretion test, it was found to improve (P < 0.05) glucose tolerance and the insulinogenic index, as compared with the respective values of these parameters before GLP-1 treatment. PMID- 9415073 TI - Evidence for a hormonal action of beta-endorphin to increase glucose uptake in resting and contracting skeletal muscle. AB - The uptake of 2-[3H]deoxyglucose, a non-metabolisable derivative of glucose, was studied in resting and contracting muscle. An isolated phrenic nerve/hemidiaphragm preparation of the mouse was used, and contractions of the muscle were elicited by electrical stimulation of the nerve. beta-Endorphin stimulated the uptake of 2-deoxyglucose in resting diaphragm muscle. The rate of uptake in the presence of the optimum concentration of beta-endorphin was similar to that in the presence of the optimum concentration of insulin over the short incubation period. beta-Endorphin also stimulated the uptake of 2-deoxyglucose in contracting muscle, but the optimum concentration of the peptide for this effect was three orders of magnitude lower than in resting muscle. The optimum concentration for insulin, however, was similar in resting and contracting muscle. An analogue of the C-terminal tetrapeptide of beta-endorphin also stimulated 2-deoxyglucose uptake, but this peptide was equally efficacious in resting and contracting muscle. It is suggested that beta-endorphin, which is released into the circulation during exercise, may have a hormonal action to increase the uptake of glucose during muscular activity. This peptide or its metabolites may be partly responsible for the insulin-independent uptake of glucose during exercise. PMID- 9415072 TI - IGF-I variants which bind poorly to IGF-binding proteins show more potent and prolonged hypoglycaemic action than native IGF-I in pigs and marmoset monkeys. AB - The relative acute hypoglycaemic potencies of IGF-I and several variants of IGF-I which bind poorly to the IGF-I binding proteins (IGFBPs) have been examined in marmosets (Callithrix jacchus) and the pig. In the marmoset study, IGF-I and des(1-3)IGF-I were compared in anaesthetised and conscious animals in a range of bolus doses from 42 to 270 micrograms/kg body weight. In the pig study, IGF-I was compared with four variants, des(1-3)IGF-I long-IGF-I, R3IGF-I and long-R3IGF-I (LR3IGF-I), which show reduced affinity for the IGFBPs as well as with insulin. Doses in the pig were 20 and 50 micrograms/kg body weight for the IGFs and 3 micrograms/kg for insulin. In each study serial blood samples were taken from 30 min before to 4 h after the bolus injection. Plasma glucose levels were decreased in a dose-responsive manner with the pig more sensitive than either the conscious or anaesthetised marmoset (maximum lowering 4.8, 3.7 and 2.5 mmol/l respectively). The IGF variants were consistently 2- to 3-fold more potent than IGF-I in each animal for lowering of plasma glucose to the nadir, with the potency reflecting the relative affinities for binding to the IGFBPs and the IGF I receptors. Thus, hypoglycaemic potency was in the order IGF-I < long-IGF-I < R3IGF-I approximately LR3IGF-I < des (1-3)IGF-I. Notably the variants suppressed plasma glucose levels over a much longer period than did IGF-I, the cumulative suppression over four hours showing an approximately 4- to 8-fold increase in the extent of hypoglycaemia. The prolonged suppression was not simply proportional to the hypoglycaemic nadir; at doses equipotent for glucose lowering, the cumulative hypoglycaemic effect for the variants in either species was about 2-fold that for IGF-I. The differential effect of the variants in the marmoset could not be accounted for by correlated changes in plasma insulin, IGF-I or IGFBP levels in plasma. Indirect effects via inhibition of glucagon, or direct effects via hepatic insulin receptors are postulated to account for the results. There was a dose-related reduction in plasma amino acids in the pig but, unlike the case for plasma glucose, only one analogue, LR3IGF-I was more potent than IGF-I. The response to LR3IGF-I was accentuated at the high dosage but on the basis of the other variants tested this effect could not be ascribed to either of the incorporated molecular variations. Despite their more rapid clearance from the circulation, variants of IGF-I which show lower affinity for binding to IGFBPs show proportionately superior potency for sustained hypoglycaemic action. Since our data were obtained in animal models of accepted relevance to humans these results point to the possible superior efficacy of the variants, especially des(1 3)IGF-I, over IGF-I for use as an adjunct to insulin treatment of hyperglycaemic conditions. PMID- 9415075 TI - Information technology, the Internet and urology. PMID- 9415074 TI - Evidence that the TRH-like peptide pyroglutamyl-glutamyl-prolineamide in human serum may not be secreted by the pituitary gland. AB - Recent studies have revealed that TRH-like immunoreactivity (TRH-LI) in human serum is predominantly pGlu-Glu-ProNH2 (< EEP-NH2), a peptide previously found in, among others tissues, the pituitary gland of various mammalian species. In the rat pituitary, < EEP-NH2 is present in gonadotrophs and its pituitary content is regulated by gonadal steroids and gonadotrophin-releasing hormone (GnRH). Hence, we reasoned that < EEP-NH2 in human serum may also arise, at least in part, from the pituitary, and that its secretion may correlate with that of gonadotrophins. Therefore, blood was simultaneously sampled from both inferior petrosal sinuses, which are major sites of the venous drainage of the pituitary gland, and a peripheral vein from seven patients with suspected adrenocorticotrophin-secreting pituitary tumours. In addition, in six postmenopausal and six cyclic women, peripheral vein blood was collected at 10 min intervals for 6 h, then a standard 100 micrograms GnRH test was performed. In the sera, TRH-LI was estimated by RIA with antiserum 4319, which binds most tripeptides that share the N- and C-terminal amino acids with TRH (pGlu-His ProNH2). In addition, LH and FSH were measured in these sera by RIA. In the blood samples taken at 10-min intervals, an episodic variation in serum TRH-LI was noted and pulses of TRH-LI were detected at irregular intervals (from one to six pulses per 6 h) in five postmenopausal and six cyclic women. In general, these pulses did not coincide with those of LH and FSH, suggesting that TRH-LI is not co-secreted with gonadotrophins. Moreover, unlike LH and FSH, serum TRH-LI did not increase during the menopause or after exogenous administration of GnRH. Whereas gonadotrophin concentrations were significantly greater in the inferior petrosal sinus than in peripheral serum, there were no differences in TRH-LI concentrations between these serum samples. In conclusion, serum TRH-LI in humans seems not to be regulated by gonadal steroids or GnRH. Moreover, serum derived directly from the pituitary contained no more TRH-LI than did peripheral serum, which suggests that the human pituitary gland does not secrete significant amounts of < EEP-NH2, and therefore does not contribute significantly to serum TRH-LI concentrations. Further research is required to identify the site of origin of < EEP-NH2 in human serum. PMID- 9415076 TI - The urologist and the Internet. PMID- 9415077 TI - Information technology, the Internet and the urologist. PMID- 9415078 TI - Urologists on the Internet: practical applications are now a reality. PMID- 9415079 TI - What makes a good web site? PMID- 9415080 TI - A British Urological Forum on the Internet. PMID- 9415081 TI - Consultation and patient information on the Internet: the patients' forum. PMID- 9415082 TI - Patient information systems. PMID- 9415083 TI - Providing expertise and access: the role of the healthcare librarian. PMID- 9415084 TI - The 'modern' journal goes online. PMID- 9415085 TI - Virtual reality in medicine. PMID- 9415086 TI - Artificial intelligence and neural networks: concept, applications and future in urology. PMID- 9415087 TI - Computer-assisted learning; experience at the Bristol Urological Institute in the teaching of urology. AB - OBJECTIVE: To report the use of a computer-assisted learning (CAL) program in the teaching of urology to medical students. SUBJECTS AND METHODS: Four CAL tutorials were developed, covering the examination of the urological patient, prostate cancer, impotence and lower urinary tract symptoms, for an initial evaluation of the use of CAL. Twenty-six third-year medical students seconded to the department for one week of urology teaching were randomized to two equal groups. One group used the CAL programs in addition to daily formal teaching, while the other group only received daily teaching. Teaching was of a standardized format, covering all aspects of urology, including the four areas covered by the CAL tutorials. Students were assessed using standardized multiple-choice questions (MCQ) at the start and end of the week's teaching. Incorrect responses were marked negatively. The content and ease of use of the CAL programs were also evaluated by a questionnaire. RESULTS: There was no difference between the groups in MCQ scores at the start of the week. The mean (SD) change in score over the week for those using the CAL tutorials was 6.0 (7.0), and for the control group was 0.9 (6.0), a significant difference (P < 0.005). Students reported the tutorials to be easy to use and of satisfactory content. CONCLUSION: The results of this study suggest that CAL programs are of benefit to students learning urology. PMID- 9415088 TI - Medicaid and long-term care for the elderly: implications of restructuring. PMID- 9415089 TI - Disability and Medicare costs of elderly persons. PMID- 9415090 TI - Pathways to disability income among persons with severe, persistent psychiatric disorders. PMID- 9415091 TI - Public-private collaboration in health and human service delivery: evidence from community partnerships. PMID- 9415092 TI - Understanding integrated rural health networks. PMID- 9415093 TI - Factors contributing to the plasticity of the extended longevity phenotypes of Drosophila. AB - A number of laboratories have constructed independently derived long-lived strains of Drosophila, each of which have similar but not identical patterns of variability in their adult longevity. Given the observed plasticity of longevity within each of these strains, it would be useful to review the operational and environmental factors that give rise to this phenotypic plasticity and ascertain whether they are common or strain specific. Our review of the more extensively analyzed strains suggests that the allelic composition of the initial genomes and the selection/transgene strategy employed yield extended longevity strains with superficially similar phenotypes but which are probably each the result of different proximal genetic mechanisms. This then offers a plausible explanation for the differential effects of various environmental factors on each strain's particular pattern of phenotypic plasticity. It also illustrates that the species has the potential to employ any one of a number of different proximal mechanisms, each of which give rise to a similar longevity phenotype. PMID- 9415094 TI - Correlational analysis in comparative gerontology: an examination of some problems. AB - Correlation coefficients are widely used in comparative gerontology. However, due to many pitfalls, only a poor knowledge can be gained from these studies in many cases. For instance, a few variables gathered from few species may be used, with no attention to possible confounding variables giving spurious correlations. This article describes some of these problems and proposes to use more thoroughly multivariate analysis in comparative gerontology. PMID- 9415095 TI - Decreased proliferative capacity and increased susceptibility to activation induced cell death in late-passage human CD4+ TCR2+ cultured T cell clones. AB - The growth characteristics in vitro of interleukin 2 (IL 2)-dependent human CD4+ alpha beta-T cell receptor-positive helper T cell clones (TCC) were studied in relation to alterations in surface phenotype, cytokine responsiveness, and susceptibility to activation-induced cell death (AICD). TCC derived from peripheral blood T cells had finite lifespans averaging 33 population doublings (PD) with a recorded maximum lifespan of 80 PD (n = 208). First analyses of the TCC were undertaken at ca. 25 PD, at which time all cells of all TCC expressed high intensity CD45RO and low intensity CD45RA, as well as high intensity CD95 (fas) and MHC class II antigens. The expression of these molecules remained elevated throughout the proliferative lifespan of the clones, but for those TCC which were initially CD28+ (the majority), the density of expression of the latter was diminished in most late-passage clones. Concomitant with this, late passage cells showed reduced responsiveness to CD28-mediated costimulation by CHO transfectants expressing human CD80 compared to early-passage cells. Additionally, the level of expression of IL 2R gamma c and IL 7R chains was commonly reduced, as was the response to IL 2 and IL 7. Despite unchanged levels of fas expression on TCC with time, late-passage cells were more susceptible to AICD than early, passage cells. These observations further document functional and phenotypic alterations in long-term cultured human T helper cells, which may be considered as biomarkers of immunosenescence. This may contribute to an improved understanding of the mechanisms underlying depressed T cell function in old age. PMID- 9415096 TI - A proton magnetic resonance spectroscopy study of aging and transformed human fibroblasts. AB - Proton magnetic resonance spectroscopy (1H MRS) has been used to monitor changes occurring during aging and transformation in human lung fibroblasts. Aging was studied in MRC-5 cells from nonsenescent (early passage) to presenescent (late passage) and senescence. Nonsenescent cells infected with SV40 virus (pretransformed) were monitored through crisis and subsequent immortalization. Aging changes were observed with one- and two-dimensional MR spectra. Cholesterol and lipid resonances were significantly increased from nonsenescent cultures to senescence. These changes could be caused by chemical or structural changes in the plasma membrane or in intracellular lipid pools. In contrast, choline levels rose from nonsenescent to presenescent cells but at senescence dropped to that of nonsenescent cells. Increased choline levels are often associated with increased cellular proliferation. After SV40 infection of MRC-5 cells there was an increase of cholesterol and lipid levels that peaked at crisis. Newly immortalized cells exhibited a drop in cholesterol and lipid to nonsenescent cell levels, but these rose again in established immortalized cells. In contrast to presensescent cultures, the levels of choline gradually increased from pretransformed to crisis phase but still continued to rise after immortalization. Thus, 1H MRS illustrates similarities in lipid behavior at senescence and crisis, whereas the choline levels are different. PMID- 9415097 TI - Nonshivering thermogenesis in adult and aged C57BL/6J mice housed at 22 degrees C and at 29 degrees C. AB - Twelve- and 28-month-old C57BL/6J male mice were housed either at room temperature of 22 degrees C or at thermoneutrality (29 degrees C) during the two months prior to experiments. Acute experiments were conducted under anesthesia, myorelaxation, and artificial ventilation. We recorded efferent electrical impulse activity in one of the sympathetic nerves innervating the interscapular brown adipose tissue in response to acute cold stimulation, when body temperature was lowered 7.5 degrees C below control level. In separate experiments we measured O2 consumption and CO2 production and calculated the nonshivering thermogenesis. We also measured the concentration of uncoupling protein in interscapular brown adipose tissue before and after three-hour cold stress. In aged mice, both sympathetic nervous activity and nonshivering thermogenesis were lower in animals housed at thermoneutrality (29 degrees C) than in mice housed at 22 degrees. Among mice maintained at 22 degrees C, but not at thermoneutrality, aged animals had greater nonshivering thermogenesis and greater cold induced concentration of uncoupling protein in the brown adipose tissue than adults. Sympathetic nervous outflow to brown adipose tissue was always greater in aged mice, regardless of the temperature of acclimation. We concluded that aged mice, housed at 22 degrees C, showed the changes in nonshivering thermogenesis associated with cold acclimation. However, an increased sympathetic outflow to brown adipose tissue in aged animals reflects an age-related elevation of the tone and responsiveness of the sympathetic nervous system. PMID- 9415098 TI - Rejuvenation of the disposable soma: repeated injury extends lifespan in an asexual annelid. AB - The disposable soma theory of senescence proposes that aging is the result of the accumulation of somatic damage with age resulting from insufficient somatic maintenance and repair. Comparative studies that show a positive correlation between longevity and DNA excision repair efficiency in mammals provide support for the theory but their validity has been questioned. A more satisfactory approach to investigate the role of somatic damage accumulation in aging would be to manipulate experimentally the levels of somatic repair and observe its effect on longevity. Here I report the results of studies in the asexual annelid Paranais litoralis where I have experimentally extended the worms' lifespan by subjecting them to repeated injury. I propose that repeated injury enhanced the normal level of repair of the worms, resulting in a rejuvenation of the soma. These results provide experimental support for the disposable soma theory of senescence. PMID- 9415099 TI - Citric acid cycle: a mainstream metabolic pathway influencing life span in Drosophila melanogaster? AB - The role of the citric acid cycle enzyme NADP-dependent isocitrate dehydrogenase (IDH-NADP) and its allele product variants in resisting the oxidative agent paraquat, was analyzed among descendants of reciprocal crosses between fast developmental time short-lived individuals (F-) and slow developmental time long lived ones (S+), in Drosophila melanogaster. Taking preadult developmental time into account, the data suggested that IDH-NADP differences in enzymatic activity between electrophoretically fast and slow allele product variants could play an important role in paraquat resistance and longevity, because individuals with slow developmental time bearing the fast electrophoretic variant of IDH-NADP ("fast" allele) were the most resistant. The fast electrophoretic variant of this enzyme is known to be the most active one and its activity is related to increased reduction of NADP to NADPH. This process could be very important for an effective balance between several pathways that use NADPH as precursor molecules and the oxidative stress defense system that uses it as an oxygen free radical reductor. We also reported a strong maternal effect on these traits, because survivors of a paraquat bioassay carrying cytoplasm inherited from slow developmental long-lived females (S+ cytoplasm) showed the highest frequency of the fast electrophoretical variant of IDH-NADP. PMID- 9415100 TI - Geographic variations of life history strategies in Drosophila melanogaster. III. New data. AB - A crucial assumption of evolutionary theories of aging is that age-specific differences of life history traits may have genetic causes. The present study focuses on the existence of such differences between eight freshly caught populations of Drosophila melanogaster. A highly significant differentiation of the populations is observed, yet it accounts for a relatively small part of the variance. It is also shown that large discrepancies may be found between the estimations of fitness based, on the one hand, on data for egg production and, on the other hand, on fertility data. This stresses the need for accurate measurements of fitness for the assessment of evolutionary theories. Finally, the results suggest that neither of the current evolutionary theories of aging is generally valid. Indeed, the age-specific differences that are found between the populations match either the antagonistic pleiotropy mechanism, or the concordant pleiotropy mechanism, or none of them. PMID- 9415101 TI - Telomeres, telomerase, and aging: origin of the theory. AB - In 1971 I published a theory in which I first formulated the DNA end replication problem and explained how it could be solved. The solution to this problem also provided an explanation for the Hayflick Limit, which underpins the discovery of in vitro and in vivo cell senescence. I proposed that the length of telomeric DNA, located at the ends of chromosomes consists of repeated sequences, which play a buffer role and should diminish in dividing normal somatic cells at each cell doubling. I also proposed that the loss of sequences containing important information that could occur after buffer loss could cause the onset of cellular senescence. I also suggested that for germline cells and for the cells of vegetatively propagated organisms and immortal cell populations like most cancer cell lines, an enzyme might be activated that would prevent the diminution of DNA termini at each cell division, thus protecting the information containing part of the genome. In the last few years, most of my suggestions have been authenticated by laboratory evidence. the DNA sequences that shorten in dividing normal cells are telomeres and the enzyme that maintains telomere length constant in immortal cell populations is telomerase. PMID- 9415102 TI - The current status of the protein error theory of aging. PMID- 9415103 TI - The uses of intraspecific variation in aging research. AB - The artificial creation of genetically long-lived populations of several invertebrate species has illustrated how researchers may take advantage of genetic variation within a species to investigate the nature and mechanisms of aging. The advantage of studying intraspecific variation is that populations will be generally similar except for the relevant demographic differences. Also, there are reasons to suspect that genetic mechanisms of aging may differ from mechanisms associated with life extension via environmental manipulations such as caloric restriction. However creating a long-lived mammalian aging model will be expensive and time consuming. An alternative approach is to seek to identify naturally occurring slowly aging populations to contrast mechanistically with a reference population. Ecologists have already noted that demographic alterations of the appropriate type are frequently associated with populations from differing latitudes, differing altitudes, or from islands. Therefore, it is likely that genetically longer- (and shorter)-lived mammal populations of the same species already exist in nature, and could potentially be exploited to inquire into the genetics and mechanisms of aging and longevity. Of particular interest is the indication that some island populations of house mice may exhibit extended longevity compared with laboratory strains. PMID- 9415106 TI - Ultrastructural study of thymic microenvironment involution in aging mice. AB - Aging involves morphological alterations of the thymus and deregulation of various immune response parameters. Altogether, these phenomena have been termed thymic involution. Using electron microscopy, we studied the morphological ultrastructure of the thymic microenvironment in aged mice. We observed cellular damages which progressively affected all the thymic stroma. At later stages (i.e., about 18-20 months old), a disappearance of the organ architecture with a drastic decrease in lymphocyte number was observed. The loss of cellular integrity of the microenvironment with lysis of cellular membranes and formation of a large and clear cytoplasmic layer engulfing a few remaining lymphocytes was noted. Extensive lipidic invasion surrounding the remaining epithelial cells grouped in nest formations and/or bordering cytics cavities was also present in these thymus from aged mice. Because the thymic microenvironment plays an important role in the "education" and functional maintenance of T cells and because the alteration of this cellular entity precedes a decline in certain immune functions, it can be suggested that membrane alterations, lack of cellular microenvironment integrity, and T cell dysfunction are correlated. PMID- 9415105 TI - Age-associated changes of cytoplasmic calcium homeostasis in cerebellar granule neurons in situ: investigation on thin cerebellar slices. AB - Mechanisms of cytoplasmic calcium homeostasis were investigated in adult and old CBA mice. The cytoplasmic calcium concentration ([Ca2+]i) was measured on fura 2/AM loaded granule neurons in acutely isolated cerebellar slices. The resting [Ca2+]i was significantly higher in senile cerebellar granule neurons, being on average 60 +/- 15 nM (n = 163) in adult and 107 +/- 12 nM (n = 129) in old neurons. The depolarization-induced [Ca2+]i transients were markedly altered in old neurons as compared with adult ones: their amplitude was smaller by about five times, the rate of rise was prolonged about two times, and the complete recovery to the resting level after the end of depolarization was about five times longer. The amplitude of calcium release from caffeine/Ca(2+)-sensitive endoplasmic reticulum calcium stores also become significantly smaller in old neurons (the amplitudes of [Ca2+]i transients evoked by 30 mM caffeine were 75 +/ 27 nM (n = 29) in adult and 25 +/- 10 nM (n = 23) in old neurons). We conclude that neuronal aging is associated with prominent changes in the mechanisms responsible for [Ca2+]i regulation. These changes presumably include lowering of voltage-gated plasmalemmal Ca2+ influx and slowing down of Ca2+ extrusion from the cytoplasm. PMID- 9415107 TI - Evolutionary patterns among measures of aging. AB - Maximum lifespan has been one of the most common aging measures in comparative studies, while the Gompertz model has recently attracted both proponents and critics of its capacity to adequately describe the acceleration of mortality in the oldest age classes. The Gompertz demographic model describes age-dependent mortality rate acceleration and age-independent mortality using the parameters alpha and A, respectively. Evolutionary biologists have predominantly used average longevity in studies of aging. Little is known about the evolutionary relationships of these measures on the microevolutionary time scale. We have simultaneously compared Gompertz parameters, average longevity, and maximum longevity in 50 related populations of Drosophila melanogaster, many of which have been selected for postponed aging. Overall, these populations have differentiated significantly for the A and alpha parameter of the Gompertz equation, as well as average and maximum longevity. These indices of aging appear to measure the same genetic changes in aging. However, in some specific population comparisons, the relationships among these measures are more complex. In a second experiment, environmental manipulation of longevity had substantially different effects from genetic differentiation, with the A parameter accounting for changes in overall mortality. The adequacy of the maximum lifespan and the Gompertz equation as indices of aging in evolutionary studies is discussed. PMID- 9415104 TI - Increased susceptibility of late passage human diploid fibroblasts to oxidative stress. AB - Three strains of human diploid fibroblasts, TIG-3, TIG-7, and MRC-5, were serially cultivated. The susceptibility of early-passage and late-passage cells at 20-30 and 60-70 population doubling levels, respectively, to hydrogen peroxide, the superoxide radical (exposure to the hypoxanthine-xanthine oxidase system), or linoleic acid hydroperoxide was examined for lactate dehydrogenase release. The susceptibility of late-passage cells to such oxidative stress was considerably enhanced compared with early-passage cells. The concentration of reduced glutathione in late-passage cells was lower by 24-44% on a per-cell number basis and by 86.0-94.5% on a per-protein-quantity basis than in early passage cells. In addition, the activity of catalase in late-passage cells was lower by 19-46% compared with early-passage cells. There was, however, no difference between the mRNA levels of catalase in early-passage and late-passage cells. The activities and mRNA levels of copper/zinc superoxide dismutase, manganese superoxide dismutase, and glutathione peroxidase in late-passage cells were all higher than in early-passage cells. These results suggest that late passage cells are more susceptible to oxidative stress than early-passage cells presumably because of decreases in cellular reduced glutathione concentration and catalase activity, and that their primary defense against oxidative stress is reduced glutathione. PMID- 9415108 TI - Deceleration of age-specific mortality rates in chromosomal homozygotes and heterozygotes of Drosophila melanogaster. AB - Age-specific mortality trajectories were estimated in mixed-sex cohorts of D. melanogaster. We studied 22,000 flies that were either second-chromosome homozygotes or heterozygotes with a randomized genetic background. Broad-sense heritabilities for longevity were estimated to be 6% for males and 9% for females. Heterozygotes lived longer than homozygotes on average, but there were exceptions to the usual heterotic pattern; in several crosses parental homozygotes had average life spans as long as that of their F1 heterozygotes. Estimated age-specific mortality rates were found to decelerate at advanced ages in both homozygotes and heterozygotes. The mortality models that best fit the data are the logistic model and the two-stage Gompertz model, both of which produce mortality trajectories that level off at advanced ages. Old-age mortality deceleration is not peculiar to inbred Drosophila. PMID- 9415109 TI - Phenotypic enhancement of longevity by environmental urea in Drosophila melanogaster. AB - The phenotypic enhancement of longevity through a variety of environmental treatments, including dietary manipulations, has been observed in various species of animals, both vertebrate and invertebrate. Elucidating the mechanisms underlying such effects has provided insights into the physiological processes contributing to the determination of lifespan. Here, we report the enhancement of longevity in adult Drosophila melanogaster maintained on food supplemented with urea, a metabolic waste product occurring naturally in Drosophila cultures, especially at high larval densities. The impact of urea on longevity is shown to be through a decrease in the age-independent parameter (A) of the Gompertz equation, rather than the age-dependent parameter (alpha), which reflects the "rate of aging." We also present evidence suggesting that the urea-induced increase in longevity is mediated exclusively through a reduction in some aspect(s) of reproduction in adult flies maintained on urea-supplemented food. PMID- 9415110 TI - Twenty-four hour growth hormone secretion in a patient with Werner's syndrome. AB - OBJECTIVE: To assess the 24-h endogenous secretory growth hormone (GH) profile and serum insulin-like growth factor-I (IGF-I) response to exogenous recombinant human growth hormone (rhGH) in a patient with Werner's syndrome. DESIGN: Blood sampling every 20 min for 24 h followed by three daily injections of growth hormone. SETTING: General Clinical Research Center. PATIENTS: Single patient with Werner's syndrome. MEASUREMENTS: Serum GH and IGF-I. RESULTS: Growth hormone pulses were absent during the 24-h monitoring period. Likewise, integrated GH concentrations were very low at 0.25 mu min/mL, and no peaks occurred after sleep onset. Following single daily administration of rhGH, serum GH and IGF-I rose. CONCLUSIONS: Our findings support previous but less extensive studies suggesting patients with Werner's syndrome have reduced growth hormone levels. Preliminary investigations using rhGH in patients with Werner's syndrome should be considered. PMID- 9415111 TI - Distinction between differentiation and senescence and the absence of increased apoptosis in human keratinocytes undergoing cellular aging in vitro. AB - We have examined the processes of differentiation and apoptosis in relation to cellular aging by using a culture system based on serial passaging of monolayer cultures of human keratinocytes in a serum-free low calcium medium. Differentiation was analyzed by cellular morphology and by the expression of keratinocyte transglutaminase. Keratinocyte cultures at all passages could be induced to differentiate into mature keratinocytes by raising the calcium concentration in the medium. Differentiation could also be induced in senescent cultures, but the process was significantly slower. Apoptosis was analyzed by cellular morphology and by the expression of tissue transglutaminase. Apoptotic cells constituted a very small proportion of the culture and no detectable change in the incidence of apoptosis occurred during serial passaging. PMID- 9415113 TI - Spatial memory and NGF levels in aged rats: natural variability and effects of acetyl-L-carnitine treatment. AB - The natural variability of behavioral performance of aged rats was used to evaluate the effect of acetyl-L-carnitine (ALCAR) on spatial learning and NGF levels in different brain areas. We used a cluster analysis procedure to subdivide the aged animals into three classes of performance (good, intermediate, and poor). These three classes were equally subdivided into controls and ALCAR treated animals in order to investigate its effect on spatial retention. The stratification of animals prior to treatment allowed us to highlight the state dependency of the action of ALCAR. The effect of the molecule in improving spatial retention was evident only in the intermediate performance group. Furthermore, the drug reduced the NGF levels in the basal forebrain of treated animals, especially in the intermediate performance group. These results suggest a performance-dependent effect of ALCAR and a nonlinear relationship between NGF levels and learning ability in aged rats. PMID- 9415112 TI - Atypical pattern of adenylyl cyclase activity in the adrenal medulla with age. AB - Concurrent reduced physiological responsiveness often occurs in those tissues in which signal transduction coupled to adenylyl cyclase is diminished with age. In the adrenal medulla, the major physiological functions are catecholamine synthesis and release, and these functions are increased with age. To examine whether the sensitivity or efficacy for adenylyl cyclase stimulation is altered with age, we determined the vasoactive intestinal peptide (VIP) and forskolin activation of adenylyl cyclase in adrenal medullary membranes from young (4 month) and senescent (24 month) male F-344 rats. Stimulation by VIP (10 microns) and GTP (10 microns) were unchanged with age. In contrast, the maximal stimulation by forskolin (100 microns) and the sensitivity for forskolin stimulation were increased with age. These data indicate that the pattern of increased adenylyl cyclase stimulation with age is comparable to that in liver tissue but atypical with respect to all other tissues in the rat. PMID- 9415114 TI - Influence of aging on gastric emptying of liquids, small intestine transit, and fecal output in rats. AB - The gastric emptying of a liquid meal, the small bowel transit, and the number of pellets and fecal output produced during a 24-h period, were studied in young (three months), adult (12 months) and old (24 months) male Wistar rats. The gastric emptying after 20 min of an intragastrically administered liquid meal containing phenol red and methylcellulose was significantly decreased in old rats. The small bowel transit after 15 and 30 min of the front of a charcoal and Arabic gum containing intragastrically administered meal was similar in the three age groups. The number of pellets and the mass of the feces produced during a 24 h period were significantly decreased with age, while the food intake was similar. The water content of the pellets was similar in the three age groups. These results show decreased gastric emptying of liquids and decreased stool mass in old rats, corresponding with the previously reported age-related changes in colonic smooth muscle contractility. Small intestinal transit was well maintained with age. PMID- 9415115 TI - Heat loss during cold exposure in adult and aged C57BL/6J mice. AB - Metabolic heat production (MHP), colonic temperature (Tco), and nonevaporative (dry) heat loss were measured in ADULT and AGED C57BL/6J male mice during cold exposure. Dry heat loss was assessed as a differential temperature (Td) between incoming and outgoing air through the chamber for indirect calorimetry. The average Td during cold exposure normalized to surface area for ADULT mice was significantly higher than that for the AGED animals (0.0618 +/- 0.0003 degree C/cm2 and 0.0553 +/- 0.0005 degree C/cm2, respectively). Linear regression analysis showed that at the same Tco AGED mice showed lower values of Td normalized to surface area, indicating that at the same body temperature they were losing less heat than ADULT animals. It was concluded that age-related decline in cold tolerance in mice is not due to a lack of ability to reduce heat loss during cold exposure. On the contrary, AGED animals had lower heat loss in comparison with ADULT. We suggest that augmentation of heat conservation mechanisms is an adaptive response to diminishing cold-induced heat production. PMID- 9415116 TI - Age-specific mortality costs of exposure to inbred Drosophila melanogaster in relation to longevity selection. AB - This article address the hypotheses that selection for early- or late-life fitness changes patterns of reproductive behavior, that this behavior may be dependent on the genetic makeup of the females, and that patterns of male mortality are strongly dependent on the type of females to which they are exposed. Flies selected for late-life reproduction and their associated stocks selected for early reproduction were exposed to flies of the opposite sex from either the same stock or a highly inbred stock. Males of both long- and short lived stocks showed an increase in early mortality when exposed to inbred females. In addition, when males were exposed to inbred females early in life they showed a lower age-specific mortality rate late in life than males exposed to females from their own stock. Interestingly, females exposed to inbred males showed a significant reduction in mean longevity. Analysis of age-specific mortality revealed that this reduction was brought about as a result of increased early mortality. Interpretation of the results from an analysis of mean longevity not only fails to identify important information--as shown from a demographic analysis of age-specific mortality--but also presents a misleading description of mortality costs. PMID- 9415117 TI - Influence of physiological factors on the age-related increase in blood pressure in healthy men. AB - The independent and collective influences of several physiological factors on the age-related increase in blood pressure in healthy men were examined. Twenty-seven younger and 25 older, mostly normotensive, healthy men were studied. Blood pressure, body fat, body fat distribution, maximal oxygen consumption (VO2max), plasma norepinephrine, dietary Na, and erythrocyte Na-K pump activity were measured. Older men showed 57% higher percent body fat, 40% higher plasma norepinephrine concentration, 14% greater mean arterial blood pressure (MAP), and 5% higher plasma K concentration than younger men (all p < 0.01). Older men showed a 38% (p < 0.01) lower VO2max, 19% (p < 0.05) lower energy intake, 18% (p < 0.05) lower Na-K pump rate constant, and a 17% (p < 0.05) lower Na-K pump rate. Group means for MAP were adjusted for combinations of plasma norepinephrine, waist:thigh ratio, VO2max, and the Na-K pump rate constant, to determine if any one variable or combination could account for the age related increase in MAP. Statistical adjustment for plasma norepinephrine, waist:thigh ratio, and Na-K pump rate constant eliminated the significant difference between MAPs for the two groups. Thus, alterations in sympathetic nervous system activity, body fat distribution, and the membrane Na-K pump activity independently contribute to the age-related increase in MAP in healthy men. PMID- 9415118 TI - Functional and quantitative measures of receptor-coupled G proteins in human mononuclear leukocytes: no change with age. AB - Aging has been associated with alterations in signal transduction for a number of neurotransmitter receptors in human tissues. Heterotrimeric G proteins play a pivotal role in postreceptor information transduction, by coupling a variety of hormone and neurotransmitter receptors to several intracellular effector functions. Developmental and age-related changes in the abundance of specific G alpha subunits have been shown in the human brain. In the present study, functional and quantitative measures of G proteins were conducted in human mononuclear leukocytes obtained from 19 healthy subjects of increasing age. Gs protein function, assessed through cholera toxin-sensitive beta-adrenergic and dopaminergic agonists induced increases in 3H-Gpp(NH)p binding capacities to membranes of mononuclear leukocytes, and Gi protein function, assessed through pertussis toxin-sensitive muscarinic agonist induced increase in guanine nucleotide binding capacity, were found to be unaltered by increasing age. Immunobloting analyses with specific polyclonal antibodies against G alpha s, G alpha i, and G alpha q subunit proteins in mononuclear leukocyte membranes obtained from the same subjects showed that the quantities of these proteins in mononuclear leukocytes were as well independent of age. Insofar as age-related alterations in cellular information transduction mechanisms in peripheral tissues are important from the etiological, diagnostic, and pharmacological aspects of age-related disorders, it is important to know that both the coupling of receptors to G proteins, the function of these proteins, and their abundance in human peripheral mononuclear leukocytes stays unaltered by the aging process. PMID- 9415119 TI - Relationship between susceptibility to protein oxidation, aging, and maximum life span potential of different species. AB - The main objective of this study was to determine whether antioxidative potential of the tissues plays a role in the aging process. Antioxidative potential was ascertained as the susceptibility of tissues to undergo protein oxidation, manifested as protein carbonyls, in response to acute oxidative stress, induced by exposure to x-rays. Brain homogenates from 22-month-old rats were more susceptible to oxidative stress than those from three-month-old rats. Brain was more susceptible to oxidative damage than the heart. A comparison of the susceptibility of brain and heart homogenates in five different species (mouse, rat, rabbit, pig, and pigeon) indicated that maximum life span potential of the species was inversely related to their susceptibility to acute oxidative stress. PMID- 9415120 TI - Age-dependent increase of indigenous DNA adducts in rat brain is associated with a lipid peroxidation product. AB - Indigenous DNA adducts (I-compounds) are considered to be a biomarker of aging tissues. Thus far, few studies have been conducted to investigate the accumulation patterns of I-compounds in the brain during aging. Particularly, identities of age-dependent I-compounds have largely remained unknown. In the current study, we have determined the amounts of I-compounds in the brains of male Fischer 344 rats at ages 1, 6, 12, 18, and 24 months using a 32P postlabeling technique. The results indicate that I-compounds increase in the rat brain age dependently from 6 to 24 months of age. Total I-adduct levels (central and upper cutouts) increase 3.5-fold from 6 to 24 months. Contrary to the results of other investigators, brains of 1-month-old rats contain the highest level of I compounds, which may be due to the hypermetabolic status during the infant period. In an effort to characterize I-compounds, different deoxynucleosides were coincubated with malondialdehyde (MDA). The results show that only deoxyguanosine (dGMP)-MDA adducts overlap with I-compounds of the rat brain DNA adducts map. A total of five dGMP-MDA adducts have been identified as responsible for I compounds in brain tissues. It is known that brain tissue contains high levels of lipids that are susceptible to oxygen free radicals and that MDA is the most abundant and genotoxic product of lipid peroxidation. The present study provides supporting evidence that lipid peroxidation and its product (MDA) may play an important role in endogenous brain DNA modification, which may partly contribute to cerebral aging and age-related degenerative disorders of the brain. The accumulation of I-compounds with aging may serve as an index of indirect oxidative damage to DNA as evidenced by the presence of MDA-DNA adducts. PMID- 9415121 TI - Oxygen consumption in adult and AGED C57BL/6J mice during acute treadmill exercise of different intensity. AB - Submaximal and maximal oxygen consumption was determined in untrained adult and aged male C57BL/6J mice during treadmill running. Each of 12-month-old (ADULT) and 24-month-old (AGED) male mice was tested on a motor-driven treadmill once at different speeds. VO2 was measured before, during, and after exercise by means of indirect calorimetry in metabolic treadmill chambers. The resting VO2 averaged 3064.67 +/- 87.71 mL/kg/h for ADULT mice and 2472.95 +/- 69.41 mL/kg/h for AGED mice. During exercise, VO2 increased linearly with work intensity (running speed): ADULT mice--from 5908.06 +/- 422.35 mL/kg/h at 3 m/min to 10861.99 +/- 174.03 mL/kg/h at 25 m/min; AGED mice--from 5217.25 +/- 263.26 mL/kg/h at 3 m/min to 7817.32 +/- 290.28 mL/kg/h at 20 m/min. Further increase of the running speed resulted in a decline of VO2 in ADULT and refusal to run in AGED mice. The results of this study demonstrated that in untrained C57BL/6J mice VO2max and maximal exercise capacity declined with age. At the same absolute and relative workloads, VO2 was lower in AGED mice. PMID- 9415122 TI - Dysregulation of IL-10 production with aging: possible linkage to the age associated decline in DHEA and its sulfated derivative. AB - Peripheral lymphoid cells isolated from the spleens and peritoneal cavities of aged mice were found to constitutively secrete the multifunctional cytokine interleukin (IL)-10 when cultured in vitro. B-Lymphocytes were implicated as the cell type responsible. Abnormal expression of this cytokine was also detected in vivo because high levels of mRNA for IL-10 were present in splenocytes freshly isolated from aged animals. In addition to the spontaneous secretion of IL-10, lymphoid cells from aged donors were hyperresponsive to exogenous stimulation with endotoxin, producing exaggerated quantities of both IL-10 and IL-6 in culture. Treatment of aged animals with dehydroepiandrosterone sulfate (DHEAS), a natural steroid, reversed the age-associated alterations in cytokine production, rendering the treated mice quite similar to mature adult controls. DHEAS treatment of aged mice also resulted in a lowering in the number of B1 cells present in the peritoneal cavity and also reduced the titers of circulating autoantibodies specific for phosphatidylcholine (PtC). Based on its wide range of biologic activities, a dysregulation in the mechanisms that control IL-10 production could be a major contributor to immunosenescence. The ability of DHEAS treatment to restore normal control over the expression of IL-10 may explain how this steroid enhances immunocompetence in aged animals. PMID- 9415123 TI - Nonshivering thermogenesis during acute cold exposure in adult and aged C57BL/6J mice. AB - In C57BL/6J adult and aged mice, housed at room temperature (22.5 +/- 1 degrees C), we measured O2 consumption and CO2 production and calculated metabolic heat production under conditions of anesthesia and myorelaxation during acute cold stimulation when body temperature was lowered 7.5 degrees C below control level. An independent group of mice was subjected to a three hour partial physical restraint at 6 degrees C and concentration of uncoupling protein (thermogenin) was measured in interscapular brown adipose tissue mitochondria at different times after cold exposure. Heat production under anesthesia and myorelaxation was about 57-66% lower than in nonanesthetized conditions, but increased significantly during cold stimulation in both age groups. Under anesthesia and myorelaxation before and during cold stimulation aged mice produced about 20% more heat than adult mice. Because in these experiments all sources of facultative thermogenesis, except nonshivering, were suppressed by anesthesia and myorelaxation, and because brown adipose tissue is the major source of nonshivering thermoproduction, we concluded that aged mice housed at room temperature have an increased thermogenesis in brown adipose tissue. This conclusion was also supported by the finding that the concentration of uncoupling protein measured in the mitochondria of brown adipose tissue after single cold exposure was significantly higher in aged than in adult mice. Therefore, we propose that the lower, cold-induced, heat production typically observed in nonanesthetized aged mice may reflect reduced thermogenic capacity of skeletal muscles. While aged mice have less brown adipose tissue than adult animals, the remaining brown adipose tissue may compensate by increasing the concentration of uncoupling protein. PMID- 9415124 TI - Quantitative comparison of in situ lipofuscin concentration with soluble autofluorescence intensity in the crustacean brain. AB - The intensity of soluble 'lipofuscin-like' autofluorescences extracted from the brain of the freshwater crayfish, Cherax quadricarinatus, was compared with the concentration of morphological lipofuscin present in sections of the same tissue. This study represents the first quantitative demonstration that these soluble autofluorescences, previously attributable to lipofuscin or fluorescent age pigment (FAP), actually bear no quantitative relationship to it at all. This result confirms previous suspicions in the literature. In some cases the intensities of these unidentified soluble autofluorescences are positive linear or curvilinear functions of organ or body weight and, therefore, may give the misleading impression that they are related to age and derived from age pigment. It is strongly recommended that researchers, particularly on aquatic species, avoid using the original biochemical assay procedure for lipofuscin and its more recent modifications. PMID- 9415125 TI - An Alzheimer's mouse at last? PMID- 9415126 TI - Plumbing the depths of crocodile blood. PMID- 9415127 TI - New diabetes-associated mutation. PMID- 9415128 TI - HIV replication. PMID- 9415129 TI - Drug discovery and delivery. PMID- 9415130 TI - Phospholipase A2 and inflammation. PMID- 9415131 TI - Patenting genes. J. Craig Venter and the Human Genome Project. AB - He often begins to answer before you have finished asking your question, but his replies are to the point. He speaks with emotion about the people he hopes to help, particularly children with fatal genetic diseases. 'I prefer to do something: the outcome of doing nothing is perfectly clear', he says. He began sequencing human genes while others were discussing how and when to do so. His methods have been called inelegant, redundant, wasteful, and too 'industrial'. Will his method fail to detect some infrequently transcribed genes? That's all right with him. He is not concerned with finding all the human genes, but rather with the fastest possible route to a useful catalog of most human genes. PMID- 9415132 TI - The molecular basis for phenotypic variability of the common thalassaemias. AB - The thalassaemias, the commonest monogenic diseases in humans, and the first to be analysed at the molecular level, show remarkable phenotypic heterogeneity. As much of this variability can be ascribed to acquired pathology resulting from complications of the profound anaemia that accompanies these diseases, it is often difficult to define the precise relationships between different mutations and their clinical manifestations. Some progress has been made, however. In this article, what has been learnt about genotype/phenotype relationships for the two common forms of thalassaemia is summarized. PMID- 9415133 TI - Classifying hepatitis C virus genotypes. AB - Hepatitis C virus (HCV) is the major aetiological agent for blood-borne non-A, non-B hepatitis worldwide. Since its discovery in 1989, at least 28 HCV genotypes have been reported, which differ by > 20% in the nucleotide sequence of the entire genome (approximately 9500 nucleotides) or the sequence of the E1 gene (576 nucleotides). Different HCV genotypes have distinct geographical distributions, and may be associated with variations in viral replication and disease-inducing activity, as well as poor response to interferons in patients with chronic hepatitis C. PMID- 9415134 TI - Molecular single-cell analysis of Hodgkin and Reed-Sternberg cells. AB - In Hodgkin's disease, the malignant Hodgkin and Reed-Sternberg (HRS) cells are present in very small numbers in the diseased tissue, thus making molecular analysis of these cells very difficult. Using micromanipulation and single-cell polymerase chain reaction (PCR), rearranged immunoglobulin genes can be amplified and sequenced from single HRS cells. Oligonucleotides chosen from the variable (V)-gene sequences identified in the HRS cells can be used as specific markers for the tumour clone. This technique will allow one to search for members of the tumour clone in various compartments of the patient's body, to follow disease progression during therapy, and to analyse stem-cell populations for contamination by tumour cells before autologous bone-marrow transplantation. PMID- 9415135 TI - Rational drug design: a multidisciplinary approach. AB - The pharmaceutical industry developed spasmodically and, in its early years, was shaped by several notable events and scientists. Paul Ehrlich, whose studies initiated the field of chemotherapy, was one of the major contributors. From early experiments on the efficacy of the biological stain methylene blue as an anti-malarial, Ehrlich realized that small molecules could be used to treat infections. Further studies identified trypan dyes and arsenical compounds as treatments of trypanosomal infection, and the arsenical compounds were found to be effective against syphilis. Ehrlich's experience of the chemical industry, in particular the chemistry of dyes, led to the realization that synthetic molecules could be medically useful and provided a powerful stimulus to pharmaceutical research. PMID- 9415136 TI - The distribution of angiotensin II type 1 receptors, and the tissue renin angiotensin systems. AB - Since its discovery, the functions of the renin-angiotensin system (RAS) have attracted a great deal of attention, and the roles it plays under normal conditions, and in disease, acquire a deepening significance with every year. In general, the RAS has been considered largely in terms of its roles in sodium and potassium homeostasis and the regulation of blood pressure. The continued acquisition of information on the distribution of angiotensin receptors, however, emphasizes that our interpretation needs to be widened, and it is now clear that angiotensin II has an array of functions in the tissues, which are unrelated to its systemic roles. This paracrine function is brought about by the existence of complete, localized tissue RASs, which respond to physiological demand independently from the systemic system. PMID- 9415137 TI - The role of cytokines and adhesion molecules in the development of inflammatory injury. AB - Clinicians are constantly challenged by patients who demonstrate the ill effects of an uncontrolled host inflammatory response. Patients with sepsis and adult respiratory distress syndrome (ARDS) are frequently encountered examples of this syndrome. Despite advances in intensive care, mortality from these syndromes remains unchanged over the past two decades. In order to gain a better understanding of this pathophysiological response and to identify more specific therapeutic targets, the techniques of molecular biology have been applied to in vivo inflammatory models. Recent data indicate that the inflammatory response is dependent on the presence of both cytokines and adhesion molecules that mediate neutrophil-endothelial cell adhesive interactions. In this article, we review our experience using a lung model of inflammation that has provided insight into the events leading to injury. Cytokines [particularly, interleukin 1 (IL-1) and tumor necrosis factor alpha (TNF-alpha)], and endothelial, as well as leukocyte, adhesion molecules appear to coordinate a cascade of interactions between leukocytes and endothelial cells, which results in tissue injury. PMID- 9415138 TI - The genetics of obesity: from genetic epidemiology to molecular markers. AB - Obesity is a highly prevalent disease that carries enormous human and economic costs in western nations. The complexity and diversity of the paths leading to an overweight or an obesity status are enormous. The etiology, causes, associated morbidity, treatment, benefits versus risks of weight loss, prevention, and other aspects of obesity are all highly complex and intimately associated with other diseases, the prevalence of which is augmented by our present way of life. This article gives a brief overview of the current status of knowledge of the genetic basis of human obesity from a genetic epidemiology, experimental genetic and molecular biology perspective. It appears likely that the susceptibility to obesity depends, to a large extent, on several autosomal genes. PMID- 9415140 TI - Monitoring germline gene therapy. PMID- 9415139 TI - Horror bacteria. PMID- 9415141 TI - Research grants. PMID- 9415142 TI - Alternative to bone-marrow transplantation. PMID- 9415144 TI - The Web goes MOO. PMID- 9415143 TI - New vaccine strategies. PMID- 9415145 TI - The web. Accessing GenBank. PMID- 9415146 TI - Keep taking the tomatoes--the exciting world of nutraceuticals. AB - The market for functional of health foods is growing steadily, and new legislation in the USA is likely to give it an enormous boost. Although the public has been aware of health foods for a long time, the latest products really merit the label 'designer foods'. There are still a number of important issues to be resolved, but industry is preparing itself for major investment in a market that could dwarf that for drugs. PMID- 9415147 TI - Hemoglobin and free radicals: implications for the development of a safe blood substitute. AB - The two major concerns in the development of cell-free hemoglobin as a blood substitute (i.e. circulatory retention and oxygen delivery) have been resolved successfully by strategic chemical or genetic modification of the protein. However, the redox reactivity of hemoglobin and its impact on the physiological processes has not been fully understood, nor has it been subject to control by design. This article reviews current research into heme-mediated toxicities that potentially constitute serious impediments to the development of a usable blood substitute. PMID- 9415148 TI - Application of antisense technology to therapeutics. AB - Antisense oligonucleotides inhibit gene expression by binding in a sequence specific manner to an RNA target. Modern nucleotide chemistry has enabled the synthesis of chemically modified oligonucleotides that are highly resistant to nuclease degradation. Among other applications, these agents are currently being evaluated as potential antiviral and anticancer drugs. However, several unsolved problems remain. Poor efficiency of delivery to cells, tissue toxicity and antisense-independent biological effects of oligonucleotides currently limit the widespread application of antisense oligonucleotides to human disease. This article reviews some of the applications of antisense oligonucleotides and discusses problems associated with these applications. PMID- 9415149 TI - Polymeric carriers for regional drug therapy. AB - The limitations of currently available drug therapies, particularly for the treatment of diseases localized in a specific organ or tissue, have encouraged scientists to consider alternative methods of drug administration to increase their specificity. This is particularly relevant in the treatment of solid tumors and localized infections, where a high rate of recurrence remains a major clinical problem, associated with inadequate drug supply to the diseased site. An alternative treatment is the local administration of the agent from a degradable polymeric delivery system implanted at the site of the disease. In this fashion, the drug depot can maintain a high local concentration at the site for extended time periods with minimal systemic distribution of the drug. The polymer carrier is degraded and eliminated from the body shortly after the drug has been released. PMID- 9415150 TI - Modelling cystic fibrosis in the mouse. AB - Animal models of human diseases are of vital importance, both for understanding basic disease mechanisms, and for developing potential treatments. Recent advances in molecular biology have helped identify the lesions underlying many genetic diseases. In conjunction with the development of techniques for manipulating the mouse embryo the possibility of mimicking these diseases in vivo has been raised. The example of cystic fibrosis illustrates this potential very well, demonstrating the usefulness of mouse models for gaining an improved understanding of the disease in humans and for the testing of new treatments. PMID- 9415151 TI - Air pollution and asthma: molecular mechanisms. AB - Air pollution, in particular that generated by road traffic, is a matter of rising public concern and has been implicated in the worsening of asthma. In this article, the evidence that air pollutants (particularly sulphur dioxide, ozone and nitrogen dioxide) can affect the airways of asthmatic patients is reviewed, and the possible molecular mechanisms that may link air pollution to increased inflammation in the airways are discussed. Airway epithelial cells may respond to oxidant pollutants by the activation of transcription factors, such as nuclear factor kappa B, resulting in increased transcription of genes for certain cytokines, such as interleukin 8 and inflammatory enzymes, such as inducible nitric oxide synthase and cyclo-oxygenase. PMID- 9415152 TI - Infectious diseases in the year of Louis Pasteur. PMID- 9415153 TI - Neural networks: a bridge between neuroscience and psychology. AB - Between the anatomy and physiology of neurons on the one hand, and the study of perception and personality on the other, there is a vast gap in our understanding. Although, today, most neuroscientists say it is axiomatic that the phenomena of mind result from the operation of the brain, they could not tell you how. PMID- 9415154 TI - Generation and utilization of synthetic combinatorial libraries. AB - The use of combinatorial chemistry is fundamentally changing the pace and scope of basic research and drug discovery. Since the introduction of synthetic peptide libraries several years ago, combinatorial chemistry has proven to be a powerful tool for the generation of immense molecular diversities of peptides, peptidomimetics and new organic compounds. This article briefly reviews methods for the generation and application of combinatorial libraries, with particular emphasis on soluble synthetic combinatorial libraries. The utility of these molecular diversities for basic research and drug discovery has been demonstrated through the identification of numerous highly active compounds such as antigenic peptides, receptor ligands, antimicrobial compounds and enzyme inhibitors. PMID- 9415155 TI - Novel therapeutic directions for Parkinson's disease. AB - Problems associated with the long-term use of L-dopa to treat Parkinson's disease have prompted considerable interest in developing alternative therapeutic strategies for this disorder. Newer approaches have included dopamine-cell transplantation, genetic manipulation of striatal cells in situ and the localized administration of trophic factors to injured neurons in the substantia nigra. These studies not only offer promise for treating Parkinson's disease, but also provide insights into properties of the nigrostriatal system that may eventually help to prevent neuronal degeneration. PMID- 9415156 TI - Cornering and catching the common protein fold. AB - Protein coding regions (genomic DNA) and cDNA sequences can be screened rapidly to help identify them. If a protein's three-dimensional fold can also be identified, then modelling can provide an estimate of the protein's structure, which can assist both in drug design and in probing structure-function relationships. Methods linking one-dimensional information, represented by the sequence, to three-dimensional information of the protein fold are key to this progression of events. The major difficulty lies with the identification of related proteins whose sequences have diverged beyond recognition, while their folds remain similar enough to confirm their descent from a common ancestor. PMID- 9415157 TI - Mechanisms in motor neurone disease: clues from genetic studies. AB - Motor neurone disease is a rapidly progressive neurodegenerative disorder, characterized by muscular weakness and wasting with fasciculation and by spasticity. While most cases are sporadic, approximately 10% are inherited in an autosomal dominant mode. Recently, mutations in the gene encoding the free radical scavenging enzyme superoxide dismutase-1 have been found to segregate with the disorder in 20% of familial cases. This is an exciting development, as free radical damage has long been implicated in the pathogenesis of motor neurone disease and it raises the possibility of novel therapeutic approaches in this otherwise fatal condition. PMID- 9415158 TI - Tissue synthesis of complement as an immune regulator. AB - Evidence is accumulating that a variety of tissues produce complement components, and that production in each tissue is differentially regulated by inflammatory cytokines. This locally produced complement could have protective or injurious actions, depending upon local circumstances. Techniques for analysing separately the contributions of local complement synthesis and complement derived from the circulation are now becoming available. We argue that an appreciation of the role of local complement synthesis may help to explain many features of organ- and tissue-specific immunological disease. PMID- 9415159 TI - Genome analysis. PMID- 9415160 TI - Homoeopathy: science or scam? PMID- 9415161 TI - Comparative protein modeling by satisfaction of spatial restraints. AB - Approximately one third of known protein sequences are related to at least one known protein structure. As a result, an order of magnitude more sequences can be modeled by comparative modeling than there are experimentally determined protein structures. A large fraction of these models has an accuracy approaching that of a low resolution X-ray structure or a medium resolution nuclear magnetic resonance structure. The number of applications where homology modeling has been proven useful is growing rapidly. PMID- 9415163 TI - Cyclosporins: immunosuppressive drugs with anti-HIV-1 activity. AB - Cyclosporin A was introduced into clinical use in the late 1970s to reduce graft rejection after organ transplantation. This drug, a cyclic undecapeptide metabolite of the fungus Tolypocladium inflatum, interferes with lymphokine biosynthesis, hence its immunosuppressive activity. Recently, it has become clear that cyclosporins are also inhibitors of HIV-1 replication. Immunosuppressive and antiviral activities are distinct functions of the cyclosporins, but both functions require an interaction of the drug with cyclophilins. PMID- 9415162 TI - Therapeutic applications of neurotrophic factors in disorders of motor neurons and peripheral nerves. AB - Research in the past few years has produced exciting progress in our understanding of neurotrophic factors. Robust effects of neurotrophic factors on neuronal survival and differentiation in animal studies have encouraged initiation of clinical trials for diseases of the human nervous system. In this article, the data for the actions of neurotrophic factors and the rationale for their use in clinical trials are reviewed. Recent data demonstrating efficacy of insulin-like growth factor 1 in amyotrophic lateral sclerosis suggest that neurotrophic factors can be used to treat neurological disease. PMID- 9415164 TI - Environmental antigens and atopic disease: underlying mechanisms and prospects for therapy and prophylaxis. AB - Natural exposure to non-pathogenic antigens, the so-called 'environmental antigens', experimentally elicits a characteristic pattern of T-cell immunity, involving selective suppression of the T-cell-dependent IgE responses that trigger allergic reactions. This 'immune deviation' results in the generation of long-lived T-cell memory, which confers active protection against allergic sensitization. Corresponding primary immune responses to environmental antigens in humans occur most frequently during infancy, and the development of allergic sensitization can be viewed as a failure of this natural immune-deviation process. In this article, potential strategies for primary prevention of allergic disease in infants involving, in particular, the exploitation of oral and mucosal tolerance to stimulate protective CD8+ T-cell-mediated immune deviation, are discussed. PMID- 9415165 TI - Unexplained illnesses not unique to Gulf War veterans. PMID- 9415166 TI - Senate FDA reform bill--good news for the pharmaceutical industry? PMID- 9415167 TI - NIH grant for Candida and AIDS research. PMID- 9415168 TI - Tolerance induction in transplantation and autoimmune diseases. PMID- 9415169 TI - Defending the use of animals to research human disease. PMID- 9415170 TI - Fetal surgery: the challenge of operating before birth. PMID- 9415171 TI - Non-viral gene therapy. AB - Gene therapy is a medical/surgical intervention currently being developed, in which genes are introduced into cells in order to treat or cure a wide variety of human diseases. The field has evolved over the past four decades, with most experimental gene-therapy studies based on the use of viruses to deliver the genes of therapeutic interest. More recently, a large number of non-viral approaches to gene therapy have emerged, yielding promising pre-clinical results, and which are currently being evaluated in early stage clinical trials. PMID- 9415172 TI - Platelet storage: efforts to extend the shelf life of platelet concentrates. AB - In transfusion medicine, platelets cannot be replaced by blood substitutes. Circulating platelets must respond quickly to changes in normal blood flow and blood-vessel injury to promote normal hemostasis. Adhesion of platelets at the site of vessel endothelial rupture is mediated through platelet membrane glycoprotein receptors. The integrity of these surface adhesion receptors and the signal-transduction pathways of activation will determine, in large part, how well a platelet functions in hemostasis. The deterioration of these systems during storage leads to a compromise of function known as the 'platelet-storage lesion'. PMID- 9415173 TI - Development and degeneration of the intervertebral discs. AB - The intervertebral discs undergo profound changes in structure and composition during development and aging. They also degenerate much earlier than other cartilaginous tissues, and in severe cases may lose function completely. The reasons for this early degeneration are unknown, but external factors, such as mechanical overload on the spine, or smoking, may be involved. However, recent work has revived interest in the importance of genetic and developmental influences on the intervertebral discs. PMID- 9415174 TI - Animal studies of cystic fibrosis. AB - Cystic fibrosis is the most common life-threatening autosomal recessive genetic disorder in Caucasian populations. It is a disease primarily of epithelial tissues, including the airway, pancreatic duct, intestine, genital tract and sweat glands. The affected gene was cloned and characterized in 1989. In the absence of an identified natural animal model of the disease, a major effort has been made to develop transgenic cystic fibrosis mice, by disrupting the gene in these laboratory animals. Such mice show many, but not all, of the symptoms of cystic fibrosis. In this article, the major past and present contributions of other animal systems to our understanding of cystic fibrosis are examined and their potential for future studies of this disease are discussed. It is intended to give the reader a broad overview of the field, exploring the usefulness of animal studies, rather than dealing more fully with specific aspects of cystic fibrosis. PMID- 9415175 TI - Is there clinical, epidemiological and molecular evidence for two forms of Crohn's disease? AB - Crohn's disease is an idiopathic chronic panenteric intestinal inflammatory disease. Data concerning the pathogenesis of, and the immune responses occurring in, Crohn's disease are often conflicting. Current therapy is empirical and either non-specifically immunosuppressive or surgically ablative in nature. Although controversial, Crohn's disease may be thought of as having two different presentations, an aggressive fistulizing form and an indolent obstructive form. This is analogous to the tuberculoid and lepromatous manifestations of leprosy. If correct, this subclassification may provide key insights into the pathogenesis and differing host immune responses in Crohn's disease and also allow the development of more rational therapies. PMID- 9415176 TI - Baboons, stem cells, facilitator cells and AIDS. PMID- 9415177 TI - Stem cells for damaged brains? PMID- 9415178 TI - Advances in tissue repair. PMID- 9415179 TI - Delivery devices and protecting the next generation from nicotine. PMID- 9415180 TI - Nitric oxide. PMID- 9415181 TI - Public understanding of medical discovery. AB - In February last year, the popular British magazine Woman's Own published an article headed 'Would you take a breast cancer test?'. It suggested that, 'women who have a family history are thought to be more at risk. Now, however, scientists have developed a blood test that can show if we're likely to get cancer. But would you want to know?'. PMID- 9415182 TI - New insights into atherosclerosis from studies with mouse models. AB - Atherosclerosis is a disease of the large arteries that is the cause of heart disease and stroke. It is a highly complex disorder with multiple genetic and environmental influences. The mouse model has proved very useful for studying atherosclerosis because genetic analysis and planned genetic modification are feasible in this organism. In this brief review, some recent findings are summarized and future prospects using mouse models to study atherosclerosis related traits are discussed. PMID- 9415183 TI - The importance of genome analysis to the drug discovery process. PMID- 9415184 TI - Aging and cancer: are telomeres and telomerase the connection? AB - There is substantial evidence for the progressive loss of the telomeric ends of chromosomes during aging, both in cell culture and in vivo. The loss of telomeres may eventually induce antiproliferative signals that result in cellular senescence. A hypothesis gaining prominence is that the activation of telomerase, a ribonucleoprotein enzyme that is important in maintaining telomere length stability, is necessary for the sustained growth of most tumors. The interrelationships between telomere shortening and aging, and how activation of telomerase may be necessary for cells to become immortal and malignant, are reviewed here. PMID- 9415185 TI - Role of endogenous enteric organisms in the reactivation of arthritis. AB - Reactive arthritis is an acute form of arthritis apparently caused by a combination of bacterial infection and genetic influences. Recent experiments using an animal model suggest that certain bacterial cell wall polymers originating from endogenous enteric bacteria may be responsible for the condition. PMID- 9415186 TI - Why men and women are incompatible. PMID- 9415187 TI - How do antidepressants work? PMID- 9415188 TI - Developing therapies for Alzheimer's disease. AB - With the greying of the population, many more people will suffer from senile dementia, placing an enormous burden on families and on health provision. Alzheimer's disease, by far the most common form of dementia, is a complex disease, whose causes are still poorly understood despite a recent mushrooming in research. Many drugs are being developed that may provide symptomatic relief or slow down deterioration, but preventive therapies still seem a long way off. PMID- 9415189 TI - Virus vector design in gene therapy. AB - The success of current and future gene therapy approaches is largely dependent upon the vector systems used to carry the therapeutic genes. Issues of efficiency, specificity and safety of gene transfer play an important role. Currently, the best vector systems available are based upon Nature's natural gene transfer systems, the viruses. This review will compare the three most common viral vector systems, retroviral, adenoviral and adeno-associated viruses, highlighting problems and issues of design, as well as suggesting the direction that future developments could take. PMID- 9415190 TI - Intrauterine programming of adult disease. PMID- 9415191 TI - Antibody-directed enzyme prodrug therapy for cancer: its theoretical basis and application. AB - Agents that can be administered systemically but that act selectively against cancer cells have been intensively sought but have thus far proved elusive. Nonselective cytotoxic drugs have the potential to eradicate cancer if they can be delivered selectively in sufficient concentration to cancer sites. In the approach described here, the cytotoxic agent is generated at cancer sites from a low-toxicity prodrug by the action of an enzyme delivered by an antibody to the cancer site. The feasibility of the approach has been demonstrated with a variety of enzyme-prodrug combinations. PMID- 9415192 TI - The molecular pathology of syndromic craniosynostosis. AB - Several monogenic disorders result in craniosynostosis, the premature fusion of skull sutures in the neonate, causing craniofacial malformation and, occasionally, neurological compromise. These malformations were initially classified on a clinical basis, but several recent reports have clarified the underlying mutations in many of these syndromes, allowing the complexity of the relationship between mutation and resultant phenotype to be viewed more clearly. This article summarizes the current situation regarding syndromic craniosynostosis, highlights the complementarity of clinical, cytogenetic and molecular approaches that have contributed to the improved understanding of the genetic basis of craniosynostosis, and considers the new challenges that have emerged. PMID- 9415193 TI - 2-Chloro-2'-deoxyadenosine (2CdA) biochemical aspects of antileukemic efficacy. AB - The studies on the metabolism and toxic mechanism of 2-chloro-2'-deoxyadenosine (2CdA, Cladribine), a new antileukemic drug, were reviewed. 2CdA, being a 2 halogenated, adenosine deaminase-resistant analogue of deoxyadenosine, is phosphorylated to the mono-, di, and triphosphate chlorodeoxy adenosine and the first step of phosphorylation is taken in the presence of enzymes, mainly kinase deoxycytidine (although in mitochondria it is phosphorylated by kinase deoxyguanosine). Triphosphate derivative of 2CdA is commonly considered to be the agent inducing cell apoptosis resulting from inhibition of ribonucleotide reductase, DNA polymerases and DNA repair. Recent studies on toxicity of 2CdA showed that the nucleoside possesses inhibitory activity against enzymes which are responsible for metabolism of deoxyadenosine, which suggests that the mechanism of toxicity by 2CdA includes a block in dAdo metabolic pathways which is very important for normal function of immune system cells. The agent under discussion and two other adenosine analogues (i.e. fludarabine and 2' deoxycoformycin) which exhibit cytotoxicity against dividing and resting lymphocytes revolutionized the treatment of indolent lymphoid malignancies (i.e. chronic lymphocytic leukemia, non-Hodgkin's lymphoma, cutaneous T cell lymphoma and hairy cell leukemia). Particularly, in the treatment of hairy cell leukemia, 2-chloro-2'-deoxyadenosine demonstrated excellent efficacy, achieved after a single 7-day course, with an acceptable tolerability profile, suggesting that cladribine is likely to be more effective than other agents recommended in this disease. Preliminary clinical data, extremely encouraging in the case of 2CdA indicate that biomolecular mechanisms of the drug cytotoxicity is worth wide presentation. PMID- 9415194 TI - Study of local anaesthetics. Part 136. Influence of auxiliary substances on the release of pentacaine from drug forms. AB - The paper deals with the formulation of a potential local anaesthetic--pentacaine into appropriate drug forms. To be more specific, the pentacaine is studies from the two aspects. First, the influence of auxiliary substances and their respective concentrations on the both onset and duration of local anaesthetic effects of pentacaine solutions. Second, the liberation of pentacaine from hydrogels and suppositories. Based on the obtained results, optimal auxiliary substances together with their optimal concentration are recommended. PMID- 9415195 TI - 2-Mercapto-N-(azolyl) benzenesulphonamides III. Synthesis of some new 2-mercapto N-(5-amino-1,2,4-triazol-3-yl) benzenesulphonamide derivatives with potential anti-HIV or anticancer activity. AB - A series of 2-mercapto-4-R1-5-R2-N-[1-R4-5-(R3-amino)-1,2,4- triazol-3-yl] benzenesulphonamides [IIa-IIg, IIIa-IIId] was obtained by the reaction of N-(1,1 dioxo-6-R1-7-R2- 1,2,4-benzodithiazin-3-yl)-N'-R3-guanidines [Ia-Ig] with some hydrazines in methanol or pyridine medium. The mechanism for the reaction has been suggested. Preliminary screening data indicated that compounds [IIIa-IIIb] exhibit moderate anti HIV activity, and compounds [IIIa-IIId] anticancer activity. Some structure-activity relationships are also discussed. PMID- 9415196 TI - Synthesis and antibacterial activities of some new arenesulfonamides and urea derivatives. AB - Reactions of 3-substituted-4-amino-4,5-dihydro-1H-1,2,4-triazol-5-ones [I] with some arenesulfonyl chlorides and aryl isocyanates were studied and the corresponding N-(3-substituted-4,5,-dihydro-1H-1,2,4-triazol-5-one-4-yl) arenesulfonamides [II-VI] and N-aryl-N-(3-substituted-4,5-dihydro-1H-1,2,4 triazol-5-one-4-yl) ureas [V, VI] were obtained. The structural assignments of new 19 compounds are based on spectral data and elemental analysis. Furthermore, compounds II-VI were tested for their in vitro antimicrobial activities. PMID- 9415197 TI - Comparison of the induction of SOS repair in Escherichia coli PQ37 and PQ243 by antineoplastic agents. AB - Six alkylating antineoplastic drugs (Cyclophosphamide, Chlorambacil, Busulfan, Melphalan. Streptozotocin and Lomustine) and two reference compounds (methyl methanesulphonate and N-methyl-N'-nitro-N-nitrosoguanidine) were investigated in the SOS Chromotest using the Escherichia coli strain PQ37 (wilde-type) and derived strain (PQ243), which carries the same markers as PQ37 and additionally tagA alkA. As a measure of the SOS induced activity induction factors of sflA::lacZ expression were determined. The strain PQ243 was more sensitive towards all compounds inducing SOS DNA repair then the strain PQ37 Cyclophosphamide was detected as negative in the strain PQ243 in the presence of an exogenous metabolic activation system. Lomustine was inactive both in the mutant strain and in the wild-type strain in the presence of S9 mix fraction as well as in the absence of it. Melphalan and Busulfan (without or with S9 mix) were shown to be positive exclusively in the strain PQ243. Based on these results, we discuss the usefulness of the strain PQ243 in the monitoring of the genotoxicity of drugs and in the genetic analysis of their mode of action. PMID- 9415198 TI - Inhibition of primates performed xenoantibody binding to pig aortic endothelial cells and its potential application to xenotransplantation. AB - IgM xenoantibodies are believed to play the most important role in the hyperacute rejection of distantly related species. The purpose of this study was to determine whether DL-Penicillamine could inactivate binding and cytotoxicity of adult baboon performed xenoantibodies to pig endothelial cells. Different concentrations of DL-Penicillamine were used to treat pooled baboon serum over various lengths of time. In order to determine the reactivity of baboon natural xenoantibodies to pig endothelial cells complement-mediated cytotoxicity assay was used. ELISA assay was used to assess IgM and IgG binding to pig endothelial cells. Subsequently DL-Penicillamine was dialyzed to determine its potential clinical application. Results indicate that baboon performed xenoantibodies class IgM and IgG bind to pig endothelial cells, but only IgM xenoantibody is cytotoxic DL-Penicillamine treatment can significantly reduce cytotoxicity and eliminate binding of IgM xenoantibodies to pig endothelial cells despite continued binding of IgG xenoantibodies to pig endothelial cells. In addition, DL-Penicillamine can be dialyzed, suggesting that it may be applicable in xenotransplantation and its toxicity can be significantly reduced by routine hemodialysis. PMID- 9415199 TI - Studies on pyrazine derivatives. XXX. Synthesis of pyrazinylamino-1,3 diazacycloalkanes of potential circulatory activity. AB - A series of pyrazinylamino-1,3-diazacycloalkanes was obtained by reaction of the corresponding substituted aminopyrazines with aliphatic amines. Selected compounds were studied with respect to their potential circulatory activity. The effect on the blood pressure, isolated rat tail artery and heart atria, and an influence on the human blood platelet aggregation were investigated. PMID- 9415200 TI - Synthesis and properties of some aminoalkanolic derivatives of xanthone. AB - A series of appropriate aminoalkanolic derivatives of xanthone III-VIII was synthesized and evaluated as potential antiarrhythmic agents. They were also tested for the effect on the cardiovascular system. The structure of the synthesized compounds was verified by IR 1H NMR, 13C NMR and mass spectral data. PMID- 9415201 TI - Application of the column liquid chromatography to the investigation of intermolecular interactions. PMID- 9415202 TI - Hyperglycemic and lipid parameters in diabetic and nondiabetic patients after acute cerebral stroke. AB - In 44 patients in acute period of cerebral stroke (35 with a normal glucose tolerance and 9 with non-insulin-dependent diabetes), the concentration of: glycohemoglobin, fructosamine, lipoprotein (a), triglycerides (TG), total cholesterol (TCh), as well as, HDL-cholesterol (HDL-Ch) and LDL-cholesterol (LDL Ch) were measured and atherogenic factors: TCh/HDL-Ch, LDL-Ch/HDL-Ch, TG/HDL-Ch were calculated. Diabetic patients in acute period of cerebral stroke showed not statistically significant increase of lipids, lipoproteins, glycohemoglobin and fructosamine concentrations, as compared with nondiabetic patients. PMID- 9415203 TI - Protoporphyrin derivatives--the use in localization of neoplasmas by titanium laser-induced fluorescence technique. AB - Examples of in vivo detection of skin neoplasmas (Mercl, melanoma) and breast carcinoma using laser induced fluorescence (LIF) and bis-1[1-alanylo-N] ethylodeuteroporphyrin (Ala-PP) are described in this study. Photosensitizer, wave titanium laser and standard CCD camera coupled to vision amplifier enable precise neoplasm imaging, Ala-PP retention monitoring and further photodynamic therapy (PDT). PMID- 9415204 TI - Synthesis and antifungal activity of some new arylidenamino compounds. AB - A series of 3-alkyl/aryl-4-arylidenamimo-4,5-dihydro-1H-1,2,4- triazol-5-ones was synthesized and characterized by elemental and spectral analysis. These compound were tested for in vitro antibacterial and antifungal activity in solid agar cultures against eight microorganisms. In these test, some of the new compounds were shown to be very potent in vitro antifungal activity against the used fungi. PMID- 9415205 TI - The effect of phenolic compounds of poplar leaves extract on cutaneous angiogenesis reaction induced in mice by human mononuclear leukocytes. AB - The influence of ether fraction of water-soluble extract of poplar leaves (Populus nigra L.) and nine phenolic acids (cinnamic and benzoic acids derivatives) on human mononuclear (MNL) leukocytes angiogenic activity was studied. Ether fraction, containing mainly phenolic acids, increased cutaneous angiogenesis induced in mice by human MNL isolated from the blood of healthy donors. Increase of angiogenic activity was also observed in case of caffeic, gallic, salicylic, ferulic and gentisic acids. Isovanillic, protocatechuic and p hydroxybenzoic acids did not influence this parameter. PMID- 9415206 TI - The pharmacoepidemiological phenomenon of age-dependent drug consumption and analysis of its regression models. AB - A randomized pharmacoepidemiological full-year, 7x realized experiment was carried out with the purpose of analyzing the age dependence of drug volumes consumed by the population, using financial and physical measuring units to express per-capita consumption. The experiment was performed for a complete metropolitan population, representing 0.5 million people. Causal age-based dependence of the drug consumption level was found in an analytical form. Subsequently, the optimum regression model describing this dependence was sought by methods of regression and correlation analysis. PMID- 9415207 TI - The substances of plant origin that inhibit protein biosynthesis. AB - Some plants were used for a long time in folk medicine as sources of anti-tumour remedies. Their effects on protein biosynthesis in vitro have been examined and described. The separate features of the peptide elongation system, isolated from tumoural cells, have been demonstrated. Some elongation factors or ribosomes have been shown to be a target site for the inhibition of protein biosynthesis caused by the substances isolated from various sources. The glycoside and caffeic acid, isolated from Melissa officinalis leaves, inhibited protein biosynthesis by direct influence the elongation factor eEF-2. The activity of this factor was also inhibited by aloin and aloeemodin. Saponin glycoside and its aglycon, isolated from Verbascum thapsiforme flowers, as well as digoxin, emetine and cepheline directly inactivated ribosomes. "Chagi" fraction, isolated from Inonotus obliquus, is responsible for the inhibitory effect caused by the aqueous tannin--less extract from this fungus. The target site for quercetin has been found to be the subunit form EF-1 alpha. It may be supposed that, the plant inhibitors of protein biosynthesis could be utilized for searching specific antitumoural preparations. PMID- 9415208 TI - Factors affecting the release of papaverine from spheres. AB - The effect of selected carriers and auxiliary hydrophilic agents on the papaverine loading and its release from spheres prepared by instilling viscous liquids into a 1% solution of calcium chloride in ethanol was studied. The most favourable loading of papaverine in potato starch, sodium alginate and sodium carboxymethylcellulose spheres was 97.4%, 95.7% and 91% respectively, and it depended on the presence of auxiliary hydrophilic agents such as polyethylene glycol 1500, polyethylene glycol 4000, glyceryl monostearate and sorbitan tristearate (Tween 65). The drug's release from spheres was diffusion controlled in accordance with the Higuchi model for the release of 80-95% of contents. It was concluded that the release rate constants depended on the drug-carrier hydrophilic agent composition. PMID- 9415209 TI - Effect of carriers and hydrophilic agents on papaverine spheres bioavailability. AB - The effect of carriers--potato starch, sodium carboxymethyl cellulose, sodium alginate, and hydrophilic agents--polyethyleneglycol 1500 (PEG 1500), PEG 4000, glyceryl monostearate (MSG), Tween 65, on bioavailability of papaverine from spheres were studied on rabbits. The data of pharmacokinetics parameters as absorption lag time, maximum plasma concentration (Cmax), time to peak concentration, absorption rate constant, elimination rate constant, bioavailability (F) depend on drug-carrier-hydrophilic agent composition. Potato starch-PEG 4000, potato starch-MSG, NaCMC-PEG 1500 spheres show best F-parameters of papaverine respectively 96%, 89%, 88%, Cmax 19.7, 21.3, 18.6 micrograms/ml reached in about 2 h. PMID- 9415210 TI - Characteristics of mutagenesis by bleomycin and adriamycin in Salmonella typhimurium: action of catalase. AB - The influence of catalase activity in adriamycin and bleomycin mutagenesis was investigated in Salmonella typhimurium TA98 and TA102, respectively. The activity of catalase in bacterial cells was inhibited by sodium azide. Mutagenicity of both drugs was not changed in bacterial cells with depressed catalase activity. PMID- 9415211 TI - Genotoxicity of hydrazino-phtalazine antihypertensive drugs assessed by an in vitro micronucleus assay. AB - The genotoxicity of antihypertensive drugs, hydralazine, dihydralazine and binazine was assessed on the base of their capacity to induce micronuclei in L929 cell line. In our previous investigations we indicated that these drugs did not induce micronuclei in bone marrow polichromatic erythrocytes in PZH SFISS mice. Present results show that hydralazine and dihydralizine can induce micronuclei in vitro and that this effect depends on time of exposure and the concentration of drug. PMID- 9415212 TI - Synthesis and pharmacological properties of 6,7-dimethoxy-1,2,3,4 tetrahydroisoquinoline derivatives. AB - Some selected 1-aryl-6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline derivatives were synthesized and evaluated as the beta-adrenoceptor agents. Some of the compounds showed a weak agonistic or antagonistic activity on these receptors. PMID- 9415214 TI - Synthesis of some N-substituted aminoalkanol derivatives of dibenzo[e.h]bicyclo[2.2.2]octane-2,3-dicarboximide with an expected beta adrenolytic activity. PMID- 9415213 TI - Derivatives of 2-mercaptobenzenesulphonamide. XIX. Syntheses, anticancer and anti HIV activities of some 2-(4-chloro-2-mercaptobenzenesulphonylimino) perhydropyrimidines. AB - Syntheses of some 2-(4-chloro-2-mercaptobenzenesulphonylimino)perhydropyrimidine+ ++ derivatives are described. The moderate anticancer and weak anti-HIV activities were observed in vitro for compounds [IX-XI]. PMID- 9415215 TI - Release of denotivir from the dispersed systems. Part I. Properties of hydrogel vehiculum and availability of active substance from intravaginal form of drug. AB - The study aimed at obtaining intravaginal drug form containing denotivir was performed. Complete deformation time, as well as pharmaceutical availability of biologically active substance suspended in liquefied vehiculum were determined in vitro. PMID- 9415216 TI - Investigations on the synthesis and properties of some N-[4-phenyl(2-pyrimidinyl) 1-piperazinyl]alkyl (hydroxyalkyl)-3,4-pyridinedicarboximides. AB - Synthesis of 2-substituted N-(4-phenyl-1 piperazinyl)propyl(butyl)- and N-2 hydroxy-3[4-phenyl(2-pyrimidinyl)-1-piperazinyl]propyl-6-methyl-3,4- pyridinedicarboximides [VI-XIV] is described. Some of them displayed depressive action on the central nervous system. PMID- 9415217 TI - Synthesis and pharmacological properties of new derivatives of 4-methyl-5-oxalo 1H-2,3,4,5-tetrahydro-1,5-benzodiazepin-2-one and beta-oxo-5-(4-methyl-1H-2,3,4,5 tetrahydro-1,5-benzodiazepin-2 -one)propionic acid. AB - 5-Ethoxalyl 4-methyl-1H-2,3,4,5-tetrahydro 1,5-benzodiazepin-2 -one [I] and 5 ethoxymalonyl-4-methyl-1H-2,3,4,5-tetrahydro-1,5-benzodiazepi n-2-one [IV] were subjected to transesterification using alcohols (propanol, butanol, pentanol) giving derivatives II-IV, VII, VIII. The derivatives I and VI were caused to undergo ammonolysis giving 4-methyl-5-oxamoyl-1H-2,3,4,5 tetrahydro 1,5 benzodiazepin-2-one [V] and 5-malonamoyl-4-methyl-1H-2,3,4,5-tetrahydro-1,5 benzodiazepin++ +-2-one [IX]. The compounds I-IX were tested towards their psychotropic activity. The preparates I, III and V had an anxiolytic action in the four-plate test. The anxiolytic properties of the compound V were confirmed in the test of the black-white box. The antagonism of all investigated compounds for toward reserpine, first of all of the preparations III, IV, V and IX, indicates an antidepressive activity dependent on a stimulation of the adrenergic system. PMID- 9415218 TI - Bicyclic [b]-heteroannelated pyridazine derivatives. Part 5. Synthesis of some triazolo[4,3-b]pyridazine and pyridazino[6,1-c]triazine derivatives as potential benzodiazepine receptor ligands. AB - 4-Substituted-tetrahydropyridazine-3,6-dione 3-hydrazones [I] reacted with strong carboxylic acids and some acyl chlorides to give the N-acylation products and/or triazolo[4,3-b]pyridazine derivatives. Derivatives of this bicyclic structure were obtained also in reactions of I with triethyl orthoformate and orthoacetate. In the reaction with phosgene, triazolo[4,3-b]pyridazine-3,6-dione, and with oxalyl chloride pyridazino[6,1-c]triazine-3,4,7-trione derivatives were obtained. PMID- 9415220 TI - Derivatives of 2-mercaptobenzenesulphonamide. XXI. Syntheses and cardiovascular effects of N-(dimethylaminoethyl)amides of 2-(perhydropyrimidin-2 yliminosulphonyl)-phenylthioalkanoic acids. AB - A series of 2-[5-chloro-4-methyl-2-(perhydropyrimidin-2-yl iminosulphonyl)phenylthio]alkanoic acids [IV-XI], their methyl esters [XII-XIX], N-(2-dimethylaminoethyl) amides, [XX-XXV] and the corresponding hydrochlorides [XXa-XXIVa] were obtained. The hydrochlorides [XXa, XXIIa, XXIVa] exhibited a comparable or stronger antiarrhythmic activity than the reference drugs, quinidine and procainamide. The influence of the tested compounds on the blood pressure and heart rate of rats as well as the acute toxicity in mice were studied. Some structure-activity relationship were also discussed. PMID- 9415219 TI - Derivatives of 2-mercaptobenzenesulphonamide. XX. Synthesis and cardiovascular effects of some 2-[4-chloro-2-(2-dialkylaminoethylthio)-5 methylbenzenesulphonylimino]- perhydropyrimidines. AB - A series of 2-[4-chloro-2-(2-dialkylaminoethylthio)-5-methyl- benzenesulphonylimino]perhydropyrimidines [IV-X] and their hydrochlorides [IVa VIIIa] were prepared. The antiarrhythmic effect of the most active compound [Va] was comparable to the reference drugs, quinidine and procainamide. The influence of the tested compounds [IVa, Va, VIa] on the blood pressure and heart rate of rats as well as the acute toxicity in mice were studied. PMID- 9415221 TI - The bioavailability of injectable, amorphous form of aztreonam. AB - Amorphos injectable form of aztreonam (BIOKTAM) was prepared. It was shown that after intramuscular or intravenous administration there are not any considerable differences in the bioavailability of aztreonam from BIOKTAM and of the drug from AZACTAM. PMID- 9415222 TI - Nitric oxide--a newly discovered chemical transmitter in human skin. AB - Nitric oxide (NO) is synthesized by many cells in the body. Low concentrations of NO have homeostatic roles in the circulation and nervous system, whereas high concentrations are biocidal, cytocidal and have immunomodulatory roles. The place of NO in the skin has recently become the focus of much attention, and this review highlights studies on the part played by NO in health and disease. PMID- 9415223 TI - Genotypic analysis of cutaneous T-cell lymphoma: a comparative study of Southern blot analysis with polymerase chain reaction amplification of the T-cell receptor gamma gene. AB - The diagnosis of early cutaneous T-cell lymphoma (CTCL) is a difficult point in dermatology. Recently, Southern blot analysis (SBA) and polymerase chain reaction (PCR) have been used to detect clonality in initial lesions in which clinical and histological findings are unspecific. Forty-one samples from 25 patients with CTCL were investigated for the presence of T-cell receptor-gamma gene rearrangement using a nested PCR technique and analysed by polyacrylamide gel electrophoresis (PAGE). Conventional SBA was also performed on 28 samples from 20 of these patients. In addition, 20 samples corresponding to patients with large plaque parapsoriasis (LPP), cutaneous B-cell lymphoma (CBCL) and eczema were analysed by PCR in the same way as were the CTCL specimens. Most of the CTCL specimens (81%) showed clonality on PCR analysis. Among patients with mycosis fungoides, 71% of initial patch lesions and 100% of plaques and tumours showed clonal disease. Clonality could be detected in three of four histologically negative post-treatment lesions. Clonal rearrangement was detected in one of three patients with LPP and in three of 10 patients with CBCL. None of the samples corresponding to patients with eczema showed positive results. SBA was significantly less sensitive than PCR in detecting clonality in CTCL patients (42% among early disease and 60% among advanced cases). The results indicate that this PCR/PAGE technique is a reliable and useful method for the detection of clonality in early skin lesions of CTCL patients and probably in the identification of silent extracutaneous involvement. PMID- 9415225 TI - Induction of mutagenic DNA damage in human fibroblasts after exposure to artificial tanning lamps. AB - There is increasing concern about the adverse health effects associated with the use of sunbeds, particularly with respect to skin photocarcinogenesis. The induction of mutagenic DNA damage is a prerequisite for the development of skin tumours, and it is well established that direct types of damage such as cyclobutane pyrimidine dimers (CPDs) give rise to mutations in tumour suppressor genes and oncogenes. In addition, ultraviolet radiation may induce indirect types of DNA damage, including oxidative products, which are also potentially mutagenic. By using specific DNA repair enzymes (T4 endonuclease V and endonuclease III) and the comet assay we have been able to detect the induction of CPDs, oxidized or hydrated pyrimidine bases and single-strand breaks in cultured human fibroblasts (MRC-5) after exposure for between 15 s and 20 min on two different commercial sunbeds containing Philips 'Performance' 100W-R or Philips TL80W/10R lamps. The ratio of endonuclease III to T4 endonuclease V sensitive sites varied substantially between the two lamps and was 3.3% and 18%, respectively. The sunbed containing the 'Performance' 100W-R lamps was as potent at inducing CPDs as was natural sunlight in fine weather. These results establish that commercial tanning lamps produce the types of DNA damage associated with photocarcinogenesis in human cells, and complement epidemiological evidence indicating the potential risk of using sunbeds. PMID- 9415224 TI - Absence of anaplastic lymphoma kinase (ALK) and Epstein-Barr virus gene products in primary cutaneous anaplastic large cell lymphoma and lymphomatoid papulosis. AB - The prevalence of the t(2;5)(p23;q35) and/or anaplastic lymphoma kinase (ALK) gene products in cutaneous anaplastic large cell (ALC) lymphomas and a potential precursor lesion, lymphomatoid papulosis (LyP), is controversial. ALK gene products, which are absent from normal lymphohaematopoietic cells, are a phenotypic marker of lymphomas carrying the t(2;5). We used in situ hybridization and immunohistology to screen 14 cutaneous ALC lymphomas, 21 cases of LyP, and one nodal ALC lymphoma associated with LyP for ALK gene products. ALK gene products were not detectable in these cases. In contrast, ALK gene products were found in a lymphonodal ALC lymphoma with subsequent extension to the skin and in t(2;5)-positive cell lines. Detection of the Epstein-Barr virus (EBV)-encoded small nuclear transcripts (EBER), and of immunoglobulin light chain transcripts served to check for the presence of cellular RNA in the tissue sections. EBER transcripts were found in scattered reactive lymphoid cells, but not in atypical or tumour cells. ALK gene expression and EBV infection seem to be a rare finding in cutaneous ALC lymphomas and LyP. This points to a molecular aetiology of primary cutaneous ALC lymphomas and LyP distinct from that of extracutaneous CD30+ lymphoproliferative disease. Detection of the t(2;5) or ALK gene products in cutaneous lymphoproliferative lesions therefore requires exclusion of extracutaneous ALC lymphoma in such patients. PMID- 9415226 TI - Genes for a range of growth factors and cyclin-dependent kinase inhibitors are expressed by isolated human hair follicles. AB - The mRNA expressions of various growth regulatory molecules in single human anagen hair follicles were analysed by reverse transcription and polymerase chain reaction. Approximately 370 hair follicles were isolated from 20 normal individuals, and 0.90 +/- 0.34 microgram (mean +/- SD) total RNA was extracted per whole hair follicle. The mRNAs of fibroblast growth factor (FGF)-1, FGF-2, FGF-5, FGF-7, transforming growth factor (TGF)-alpha, TGF-beta 1, hepatocyte growth factor, insulin-like growth factor (IGF)-I, tumour suppressor gene p53 and high sulphur protein were detected in most or all of the examined hair follicles per target gene. In contrast, none of the mRNAs of FGF-3, FGF-4, FGF-6, FGF-9 and IGF-II was detected, and those of TGF-beta 2 and TGF-beta 3 were detected in only a limited number of the examined hair follicles. Among cyclin-dependent kinase inhibitors, the mRNAs of p21waf1/cip1 and p27kip1 were expressed in almost all the hair follicles, while those of p15INK4B and p16INK4A were not detected. These results suggest that both positive and negative factors for the proliferation and differentiation of follicular epithelial cells coexist in a human anagen hair follicle. PMID- 9415227 TI - A controlled study of the effects of RU58841, a non-steroidal antiandrogen, on human hair production by balding scalp grafts maintained on testosterone conditioned nude mice. AB - Human hair growth can be monitored for several months after the transplantation of scalp samples from men with androgen-dependent alopecia on to female nude mice. Hair production from balding sites has been shown to be inhibited in testosterone-conditioned nude mice. We used this recently reported model to study the effect of a new non-steroidal antiandrogen-RU58841-on human hair growth. Twenty productive scalp grafts from balding men were maintained for 8 months after grafting on to nude mice, and hair production was monitored monthly for 6 months. All mice were conditioned by the topical application of testosterone (testosterone propionate, 300 micrograms in 10 microL; 5 days/week) on the non grafted flank. The scalp samples were divided equally according to the estimated hair production potential, which was based on the amount of hair present on the scalp samples before grafting. Each of the two equal groups of grafts was further allocated at random to be treated topically (5 days/week) with blinded solutions of either RU58841 1% in ethanol, or ethanol as a control. Twenty-eight active follicles appeared on the 10 control grafts. Among them only two follicles (7%) initiated a second hair cycle. However, the 10 RU58841-treated grafts bore a total of 29 active follicles, and eight of them (28%) showed a second cycle. The values for the linear hair growth rates (LHGR) were significantly (P < 0.04) higher in the RU58841-treated group. Recycling and increased LHGR indicate a positive action for RU58841 on human hair growth from balding samples grafted on to testosterone-conditioned nude mice, and encourage a clinical trial to evaluate its potential in the treatment of androgen-dependent alopecia. PMID- 9415228 TI - Expression of bcl-2 antagonist bak in inflammatory and neoplastic skin diseases. AB - Bak (bcl-2 homologous antagonist/killer) is a proapoptotic member of the ever expanding bcl-2 gene family, a recently described category of oncogenes that is critical for the regulation of programmed cell death. We investigated the expression of bak in several inflammatory and neoplastic skin diseases in comparison with normal skin. Immunohistochemical analysis revealed positive bak staining in epidermal keratinocytes of normal skin, with the granular layer being stained slightly more strongly than the basal and spinous layers, and in psoriasis vulgaris, lichen planus, actinic keratosis, keratoacanthoma and squamous cell carcinoma. We demonstrated the expression of bak in the follicular infundibulum in contrast to the outer root sheath of the lower follicle, which showed only negative to weak bak expression. Seventeen of 20 basal cell carcinomas examined showed negative immunostaining for bak, and the remaining three basal cell carcinomas showed only partial weak positivity, mainly in the palisading layers of some tumour formations. Immunoblot analysis using cultured normal human epidermal keratinocytes revealed the presence of bak protein in both undifferentiated and differentiated keratinocytes. The results of our study suggest that the loss of bak expression, in conjunction with the previously reported overexpression of bcl-2, might contribute to the pathogenesis of basal cell carcinoma. PMID- 9415229 TI - Expression of stem cell factor in basal cell carcinoma. AB - Stem cell factor (SCF) distribution in basal cell carcinomas (BCCs) was examined by immunohistochemistry. Eighteen BCCs (11 nodular, three superficial, two cystic, one adenoid and one morphoeic type) showed positive expression of SCF in the tumour islands. The centre of the tumour island was strongly positive in nodular, superficial and morphoeic types. In cystic BCCs, SCF-positive tumour cells were also located in the peripheral lesion around the cystic space. SCF was also detected on fibroblast-like cells and mast cells in the stroma. SCF was positively stained within the upper keratinocytes in the overlying epidermis, more strongly as compared with normal skin. The mast cell number (mean +/- SD) was significantly increased in the peritumoral stroma (85.7 +/- 28.3/mm2) compared with normal skin (32.1 +/- 4.2/mm2) (P < 0.005). SCF was also positive in the tumour nests of four cases of tricho-epithelioma, in which fibrosis of the surrounding stroma was found histologically. This study demonstrates that abundant SCF produced by the tumour cells may account for the increased number of stromal mast cells, which induce fibroplasia of the surrounding stroma. PMID- 9415230 TI - Cell renewal, cell differentiation and programmed cell death (apoptosis) in pilomatrixoma. AB - Pilomatrixoma is a benign tumour of the cutaneous adnexa. Histologically, pilomatrixoma comprises masses of immature basophilic cells, small numbers of polygonal squamoid cells, few transitional cells, and clusters of 'shadow cells'. The mechanism leading to the formation of shadow cells is still unknown. Skin biopsy specimens of pilomatrixoma (n = 15) were studied histologically, immunohistologically, and by applying the in situ end-labelling technique. The basal layer of the basophilic cells induced most of the proliferating cells with high expression of bcl-2 and cytokeratin 19. The overlying basophilic cells showed a negligible mitotic activity, a high significant accumulation of p53 protein, and a heterogeneous, but progressive loss of bcl-2 and cytokeratin 19. They developed either into squamoid cells or into transitional cells. The squamoid cells were characterized as differentiated cells resembling mature keratinocytes of stratified mucosa. The transitional cells could be shown to represent apoptotic cells proceeding to shadow cells. The data suggest that apoptosis is the main mechanism leading to the development of the dead shadow cells and is most probably responsible for the banal biological behaviour of pilomatrixoma. Apart from that, pilomatrixoma represents a suitable biological model to study apoptosis in humans. PMID- 9415231 TI - Glutathione efflux associated with a low gamma-glutamyl transpeptidase activity in human melanoma cells. AB - The cellular concentration of reduced glutathione (GSH) modulates the sensitivity of human melanoma cells to alkylating drugs in vitro. To investigate whether the membrane-associated enzyme gamma-glutamyl transpeptidase (gamma-GTP) involved in GSH breakdown was expressed in melanoma cells, the enzymatic activity of gamma GTP as well as the secretion of GSH were measured in human melanoma cells from four different cell lines (Me8, JUSO, GLL19, Swift). All the cells showed low gamma-GTP activities (0-1 mU/mg protein) and released GSH in culture supernatants at significant rates. After incubation for 24 h in growth medium containing 0.1 mmol/L cystine, the levels of GSH in supernatants ranged from 56 to 111 nmol GSH/mg protein. The GSH metabolism of melanoma cells was also evaluated by measuring the levels of the melanogenesis intermediate 5-S-cysteinyldopa under different experimental conditions. The results of these experiments suggest that melanoma cells have a low ability to metabolize the tripeptide GSH, which appears to be responsible for GSH secretion and accumulation in culture supernatants. PMID- 9415232 TI - Silver aids healing in the sterile skin wound: experimental studies in the laboratory rat. AB - Incisional wounds 15 mm long were induced surgically in the back skin of young adult Wistar rats. They were sutured and used as an experimental model in the therapeutic evaluation of daily applications of 0.5 mL of silver nitrate (SN) at 0.01, 0.1 or 1.0% w/v aqueous solution, or 0.5 g silver sulphadiazine (SSD) over a 10-day period. Control wounds received deionized water only. The silver preparations were not toxic but SN did stain the hair and superficial layers of the stratum corneum. The wounds remained microbiologically clean. Wounds exposed to SN (0.1 or 1.0%) or SSD healed more rapidly than controls. From about the fourth day of treatment, we noted a more rapid exteriorization of sutures, improved wound closure and an earlier loss of scabs and wound debris. Silver treatment appeared to reduce the inflammatory and granulation tissue phases of healing and enhance epidermal repair. Silver from SN was deposited as silver sulphide in extrafollicular hair shafts and superficial aspects of the skin and wound debris but not at deeper levels. Silver uptake was four-fold higher in damaged skin than in intact tissue. SSD was absorbed by intact and wounded skin but the silver did not precipitate as silver sulphide and its localization in the tissue is not known. Uptake of silver from SN or SSD was associated with changes in the concentrations of zinc and calcium in the skin. Zinc levels were depressed during the inflammatory and proliferative phases of healing and then increased. Zinc concentrations had normalized by 10 days when wound healing was achieved. Calcium levels remained higher than normal throughout the observation period. The mechanism of action of silver in advancing wound healing in the rat is unclear. Its ability to reduce the inflammatory and granulation phases of healing, and to invoke metallothionein production and influence metal ion binding are possibly important. PMID- 9415233 TI - Selectivity of protoporphyrin IX fluorescence for condylomata after topical application of 5-aminolaevulinic acid: implications for photodynamic treatment. AB - To examine the potential of using photodynamic therapy (PDT) in condylomata, we studied the distribution and kinetics of protoporphyrin IX (PpIX) formation in condylomata acuminata and adjacent normal skin after topical application of 5 aminolaevulinic acid (ALA). PpIX fluorescence spectra were measured hourly in vivo after ALA application. After gross fluorescence imaging, the lesions were biopsied, and fluorescence microscopy was performed. All three PpIX fluorescence detection modalities suggested selectivity of PpIX formation in condylomata after topical ALA application. In 17 of 25 condylomata, there was significantly greater fluorescence compared with adjacent normal skin. The greatest lesional to normal skin fluorescence ratios occurred after 2 h. The most likely mechanism for increased lesional PpIX formation in condylomata is enhanced stratum corneum permeability. Based on our results, ALA/PDT is a potential field therapy for condylomata. PpIX fluorescence imaging after ALA application may also be useful for localizing condylomata prior to treatment. PMID- 9415234 TI - The variable response of plaque psoriasis after a single treatment with topical 5 aminolaevulinic acid photodynamic therapy. AB - We have investigated the clinical response of 22 patients with plaque psoriasis to photodynamic therapy using topical application of 5-aminolaevulinic acid followed by a single exposure to broad-band visible radiation. Light doses in the range 2-16 J/cm2 delivered at dose of 10-40 mW/ cm2 resulted in a variable clinical response. Seven (35%) patients showed clearing of psoriasis at some treated sites. The intensity of protoporphyrin IX fluorescence was recorded before, during and after treatment. Pre-illumination fluorescence intensity varied considerably between sites on the same patient and between patients. Protoporphyrin IX fluorescence recovered and persisted after treatment for up to 14 days and became higher than preillumination levels at 25% of sites. The rate of protoporphyrin IX photo-oxidation during treatment was proportional to both initial fluorescence intensity and incident light dose rate and was almost complete after 16 J/cm2. We have defined the photodynamic dose as the product of time-dependent protoporphyrin IX concentration and light dose and demonstrated that only in those patients who showed clearance of psoriasis was there a relationship between photodynamic dose and clinical response. Discomfort ranged from stinging through to burning, was significant in some patients and tended to be more severe with increasing photodynamic dose but was not predictable. Efficacy may improve by achieving consistent protoporphyrin IX levels or by using multiple treatments. PMID- 9415235 TI - Five years' experience of treating port wine stains with the flashlamp-pumped pulsed dye laser. AB - During the last 5 years, 640 patients had treatment to their port wine stains (PWS) with a flashlamp-pumped pulsed dye laser. One hundred and fifty-six patients have been discharged for varying reasons, of which 59 (38%) achieved excellent (at least 75%) lightening of their birthmark. Of the remaining patients, those who attended the clinic for the sixth and 12th time for treatment were also assessed to determine the degree of fading achieved in the port wine stain. Our findings confirm that flashlamp-pumped dye laser treatment is safe and effective for the treatment of PWS and that complications are rare. However, the degree of fading achieved is variable and often unpredictable. Fifty-two per cent of facial lesions of different colours achieved over 75% fading as against 18% of non-facial lesions. Sixty-four per cent of those over the age of 50 years had an excellent response whereas only 19% of those below the age of 5 years were able to achieve a similar result. The colour of the port wine stain was found to be of no prognostic value. A search for an accurate and non-invasive method to predict the outcome of flashlamp-pumped pulsed dye laser therapy for PWS is warranted. PMID- 9415236 TI - Quality of life in patients with psoriasis: the contribution of clinical variables and psoriasis-specific stress. AB - The purpose of this study was: (i) to examine the impact of the clinical severity, anatomical location and treatment of psoriasis on patients' quality of life, and (ii) to investigate the effects of perceptions of psoriasis-related stress on patients' physical and mental health and on areas of disability in everyday life. All patients (n = 204) attending a psoriasis specialty clinic were invited to complete a multidimensional quality of life assessment comprising the Psoriasis Disability Index (PDI), the SF-36 Health Survey and the Psoriasis Life Stress Inventory (PLSI). Results (n = 150) indicated that overall clinical severity of psoriasis as assessed by the Psoriasis Area and Severity Index, and duration of psoriasis, were unrelated to impairment in any areas of quality of life. Anatomical location (social visibility) of psoriasis was associated with self-report of poor physical health (P = 0.01), and there was a modest association with patients' mental health (P = 0.04); however, anatomical location of psoriasis was not significantly associated with self-reported disability in everyday life, or stress scores. Patients who were classified as more reactive to the stress associated with psoriasis (78% of the sample) were functioning less well in terms of their mental health (P = 0.001) and also experienced significantly more disability in all areas of everyday life (P = 0.001). Differences in method of treatment for psoriasis did not significantly affect scores on the psoriasis-specific (PDI; PLSI) or generic (SF-36) quality of life measures. A multiple regression analysis demonstrated that stress resulting from anticipating other people's reactions to their psoriasis contributed more to the variance in patients' disability in everyday life than any other medical or health status variable. The results support the importance of assessing the effects of stress in patients' adjustment to their condition and may indicate a role for adjunctive psychological stress management training for a significant number of patients with psoriasis. PMID- 9415237 TI - An audit of the completeness of non-melanoma skin cancer registration in Greater Glasgow. AB - The incidence of basal cell carcinoma and squamous cell carcinoma (non-melanoma skin cancer) is increasing in the U.K., and the importance of this has been recognized in the 'Health of the Nation' target of halting the annual increase in the incidence of skin cancer by the year 2005. An accurate assessment of incidence is necessary both in meeting this target and in planning skin cancer services. We have examined the ways in which basal cell carcinoma and squamous cell carcinoma are diagnosed and treated in Greater Glasgow and have determined how many of these tumours are, recorded by the West of Scotland Cancer Registry. Our results show that there is under-registration of both basal cell carcinoma and squamous cell carcinoma. Overall, 39 of 127 basal cell carcinomas (31%; 95% confidence interval [CI] 23-39%) and 11 of 25 squamous cell carcinomas (44%; CI 26-63%) were not registered by the cancer registry. We also showed that dermatologists rarely treat clinically suspicious tumours without obtaining pathological proof of the diagnosis. Accurate data collection by selected representative cancer registries is suggested as a possible solution. PMID- 9415238 TI - Susceptibility of Malassezia furfur subgroups to terbinafine. AB - Malassezia furfur, the fungus causing pityriasis versicolor, has been reported to be sensitive to terbinafine in vitro but although topical therapy is effective in the treatment of pityriasis versicolor, oral therapy is not. This phenomenon was investigated by determining the susceptibility to terbinafine of different M. furfur subgroups in vivo (during topical or oral application) and in vitro. All M. furfur subgroups were suppressed (approximately 10-fold) by topical terbinafine. Oral treatment resulted in no significant suppression of cutaneous M. furfur populations with the exception of a single subgroup (A), which was reduced to undetectable levels on the skin of eight of 10 patients receiving oral terbinafine. Isolates of subgroup A were also markedly more susceptible to terbinafine in laboratory tests. The importance of the recognition of distinct subgroups within the cutaneous lipophilic yeasts when evaluating their antifungal susceptibility and their role in disease is discussed. PMID- 9415239 TI - A vesicular variant of bullous pemphigoid with autoantibodies against unidentified 205- and 150-kDa proteins at the basement membrane zone. AB - We describe a 75-year-old man who developed a vesicular variant of bullous pemphigoid with a distinctive result of immunoblot analysis. Characteristic symptoms consisted of vesiculopapular eruptions with erythematous patches on the arms and legs, many of which fused to form irregularly outlined areas of erythema varying in size. Direct immunofluorescence revealed a linear deposition of IgG at the basement membrane zone of the skin, and indirect immunofluorescence detected circulating IgG autoantibodies at a titre of 1:160, which reacted with the antigens located on the epidermal side of skin split with 1 mol/L NaCl. Immunoblot analysis using epidermal extracts demonstrated the presence of IgG antibodies directed to 150, 205, 240 and 280 kDa proteins as well as to the 180 kDa bullous pemphigoid antigen (BPAG2). All antibodies eluted from nitrocellulose membrane imprints of individual bands with molecular weights of 150, 180 and 205 kDa were found by indirect immunofluorescence to react with the basement membrane zone, whereas those eluted from the bands with molecular weights of 240 and 280 kDa did not. These findings suggest that antibodies directed not only to the 180 kDa BPAG2, but also to 150 and 205 kDa proteins, are involved in the pathogenesis of bulla formation in this patient. PMID- 9415240 TI - Autoantibody formation against a 190-kDa antigen of the desmosomal plaque in pemphigus foliaceus. AB - We report changes in the antigen recognition pattern of sera from two pemphigus foliaceus patients with a long-term follow-up. The patients' sera were analysed by immunoblotting using different antigenic sources: cultured human keratinocytes, bovine tongue epithelium and a recombinant protein corresponding to the C-terminal end of the 230-kDa bullous pemphigoid antigen. While initial serum samples reacted exclusively with the 160-kDa desmoglein 1, the later sera reacted both with desmoglein 1 and a 190-kDa antigen immunolocalized to the desmosomal plaque, previously demonstrated to be recognized by sera of some patients with paraneoplastic pemphigus. IgG subclass analysis further showed that antidesmoglein 1 antibodies were of IgG1 and/or IgG4 subclasses, while anti-190 kDa antibodies were IgG3. The patients were free of malignancy. PMID- 9415241 TI - Anti-epiligrin (laminin 5) cicatricial pemphigoid and lung carcinoma: coincidence or association? AB - Anti-epiligrin cicatricial pemphigoid (CP) is a rare subset of CP in which patients have IgG autoantibodies directed against the dermal side of skin split by 1 mol/L NaCl. The antibodies react against epitopes in the lowermost portions of the lamina lucida, and immunoprecipitate epiligrin (laminin 5) in human keratinocyte extracts. We report a patient with this uncommon form of CP, who, following an 8-year period of well-controlled disease, experienced a severe flare in symptoms coincident with development of lung carcinoma. This raises the possibility of an association, in this case, between anti-epiligrin CP and lung carcinoma. PMID- 9415242 TI - Psoriasis vulgaris coexistent with epidermolysis bullosa acquisita. AB - Autoimmune bullous diseases, such as bullous pemphigoid or pemphigus vulgaris, occasionally develop in psoriatic patients. In addition, a novel subepidermal bullous disease with autoantibodies against a lower lamina lucida antigen of 200 kDa has recently been reported in association with psoriasis. We describe here a patient with psoriasis vulgaris who developed epidermolysis bullosa acquisita (EBA). Direct immunofluorescence revealed linear deposition of IgG and C3 at the basement membrane zone. The patient's serum bound to the dermal side of salt split normal human skin. However, immunoblot analysis demonstrated that the patient's serum reacted with an EBA antigen of 290 kDa. EBA should be included in the list of autoimmune diseases associated with psoriasis vulgaris. PMID- 9415243 TI - Malignant melanoma and granulomatosis. AB - Sarcoidosis or granulomatous reactions have rarely been reported in association with malignant melanoma (MM). We describe seven patients who presented with both granulomatous disease and MM, and discuss the physiopathological and prognostic significance of this association. In three patients, the granulomatosis was diagnosed as true sarcoidosis and in one patient, as tumour-associated granuloma. In three cases, designated here as atypical tumour-associated granulomatoses, the presence of clear-cut pulmonary granulomatous nodules was typical neither for sarcoidosis nor for tumour-associated granuloma and was highly suggestive of melanoma metastases. Mediastinal lymphadenopathy was present in every patient. In all seven patients, the question of mediastinal or pulmonary involvement or relapse of the MM was raised, but could be confirmed in only one patient. MM can be associated with granulomatous disease. Knowledge of this association has implications in the management of patients with MM. PMID- 9415244 TI - Squamous cell carcinoma, malignant melanoma and malignant fibrous histiocytoma arising in burn scars. AB - The most common cancer arising from an old burn scar is squamous cell carcinoma, while malignant melanoma is rare. There are only four reports in the literature on the combination of the more common tumour with a malignant melanoma and only two cases of malignant fibrous histiocytoma alone. This paper reports the first occurrence of the combination of all three types of cancer in burn scars of the same patient. The factors that promote malignant transformation, such as immunological factors and karyotype, are discussed. We also performed an immunohistochemical study using anti-p53 antibodies. PMID- 9415245 TI - Necrotizing granulomatosis of the breast. AB - We describe a case of necrotizing granulomatosis of Wegener's type involving the breasts of a 40-year-old man. There were no signs of generalized disease. Involvement of the breast is rare in Wegener's granulomatosis (WG). To date, 17 cases have been reported, and all were women. They predominantly presented with a unilateral breast mass, and mammary malignancy was the principal concern. In the majority of cases, breast lesions of WG have been a presenting sign of, or preceded, disseminated disease. Our patient is unusual in that the necrotizing granulomas developed as an isolated finding in a site remote from those usually affected by WG, and, as far as we are aware, represents the first case of Wegener's type granulomatosis involving the male breast. PMID- 9415246 TI - Hypocomplementaemic urticarial vasculitis associated with Jaccoud's syndrome. AB - We report a 33-year-old Japanese man diagnosed as having hypocomplementaemic urticarial vasculitis at the age of 21, who subsequently developed Jaccoud's syndrome. Although Jaccoud's syndrome has been most frequently seen in patients with systemic lupus erythematosus, an association with other diseases has occasionally been described. Jaccoud's syndrome clinically shows joint deformities similar to rheumatoid arthritis, and needs to be differentiated from it. Patients with hypocomplementaemic urticarial vasculitis may develop Jaccoud's syndrome. PMID- 9415247 TI - Acute generalized exanthematous pustulosis following oral nystatin therapy: a report of three cases. AB - We report three cases of acute generalized exanthematous pustulosis (AGEP) following oral administration of nystatin. All cases showed similar clinical features and histopathological findings, and a delayed-type hypersensitivity to nystatin could be demonstrated in patch and prick testing. Drug eruptions to nystatin are extremely rare, and, to our knowledge. AGEP has not been reported previously. PMID- 9415248 TI - Destructive herpetic whitlow in AIDS: report of three cases. AB - Herpes simplex virus infection in immunocompromised individuals, including AIDS patients, is characterized by its tendency for atypical presentations and unusual locations, often resulting in delayed diagnosis and treatment. Three HIV-infected patients who developed prolonged cutaneous lesions of the fingers are presented. These lesions were unmodified by previous antibiotic treatment, and rapidly progressed to the complete destruction of nail structures in two patients. Viral culture confirmed the diagnosis of herpetic whitlow in all cases, and treatment with oral acyclovir resulted in complete recovery. Surgical treatment was not necessary. PMID- 9415249 TI - Verruciform xanthoma in association with a vulval fibroepithelial polyp. AB - We report the clinical, light microscopic, immunohistochemical and ultrastructural features of a verruciform xanthoma that developed in association with a vulval fibroepithelial polyp. To our knowledge, this is the first time that this association has been reported. Immunohistochemical findings confirmed that the xanthoma cells were of a monocyte/macrophage lineage. In addition to typical histological characteristics, prominent vascular ectasias were detected in the deep dermis at the periphery of this lesion. The ectasias may play a part in pathogenesis. PMID- 9415250 TI - Epidermal naevi associated with trichilemmal cysts and chromosomal mosaicism. AB - We present the case of a 29-year-old woman who has had a widespread epidermal naevus since birth, located on the left side of her body, upon which multiple trichilemmal cysts developed several years later. The karyotype of peripheral blood lymphocytes revealed chromosomal mosaicism 46, XX, t (1;9) (p36:q34). The possible pathogenic mechanisms include paracrinopathy, germinal or somatic mutation in the multipotential embryo cells, or a combination of the two. PMID- 9415251 TI - Bullous pemphigoid localized to the mouth. PMID- 9415252 TI - Bullous pemphigoid localized to the mouth. PMID- 9415253 TI - Bullous pemphigoid at sites of trauma. PMID- 9415254 TI - Severe bullous pemphigoid responsive to pulsed intravenous dexamethasone and oral cyclophosphamide. PMID- 9415255 TI - A family with actinic prurigo and polymorphic light eruption. PMID- 9415256 TI - Pellagra associated with amyloidosis secondary to multiple myeloma. PMID- 9415257 TI - Fulminating rosacea conglobata (rosacea fulminans) and ulcerative colitis. PMID- 9415258 TI - Acquired progressive kinking of the hair in a prepubertal boy. PMID- 9415259 TI - Erysipelas melanomatosum. PMID- 9415260 TI - Reinfection with lymphocutaneous sporotrichosis. PMID- 9415261 TI - Oropharyngeal lichen planus associated with interferon-alpha treatment for mycosis fungoides: a rare side-effect in the therapy of cutaneous lymphomas. PMID- 9415262 TI - Acute generalized exanthematous pustulosis due to diltiazem: confirmation by patch testing. PMID- 9415263 TI - Roaccutane and wax epilation: a cautionary tale. PMID- 9415264 TI - Phototherapy with low intensity laser irradiation for a chronic radiation ulcer in a patient with lupus erythematosus and diabetes mellitus. PMID- 9415265 TI - Another case where bigger is not better. PMID- 9415266 TI - Growth hormone: a newcomer in cardiovascular medicine. PMID- 9415267 TI - Calcium sensitisers: mechanisms of action and potential usefulness as inotropes. PMID- 9415268 TI - Transesophageal versus intracoronary Doppler measurements for calculation of coronary flow reserve. AB - OBJECTIVE: The present study was performed to compare coronary flow reserve by transesophageal Doppler echocardiography and intracoronary Doppler flow wire measurements in patients with LAD disease. METHODS: 17 patients with various degree of LAD stenosis were studied. Intracoronary LAD Doppler measurements were performed at baseline and after intracoronary injection of 18 micrograms adenosine. Transesophageal coronary sinus and LAD Doppler measurements were performed at baseline and after intravenous dipyridamole (0.6 mg/kg/5 min). Coronary flow reserve was calculated as the ratio of hyperemic to baseline average peak velocities. RESULTS: Coronary flow reserve was 2.44 +/- 0.62 and 2.19 +/- 0.76 for proximal and distal intracoronary measurements and was 2.25 +/- 0.64 and 1.74 +/- 0.63 for transesophageal LAD- and coronary sinus measurements. Proximal intracoronary flow reserve significantly correlated with transesophageal coronary sinus (r = 0.73, p < or = 0.001) and LAD (r = 0.70, p < or = 0.005) measurements, whereas distal intracoronary flow reserve only correlated with transesophageal coronary sinus flow reserve (r = 0.56, p < or = 0.02). Receiver operating characteristic curve analysis demonstrated similar diagnostic accuracy of all applied techniques for detection of a significant LAD stenosis. CONCLUSIONS: Coronary flow reserve by both transesophageal techniques correlated with intracoronary Doppler flow wire measurements, however considerable discrepancies may occur in the individual patient. PMID- 9415269 TI - Comparative analysis of plasminogen activator inhibitor-1 expression in different types of atherosclerotic lesions in coronary arteries from human heart explants. AB - OBJECTIVES: Clinical manifestations of coronary heart disease result primarily from the progressive development of atherosclerotic plaques and subsequent thrombus formation: processes which may be accelerated by an enhanced expression of plasminogen activator inhibitor (PAI-1) in the vessel wall. In the present study, content and expression of PAI-1 were comparatively analyzed in human coronary arteries in relation to the presence and severity of atherosclerotic lesions. METHODS: Segments of coronary arteries obtained from heart explants (n = 15) were classified by the presence and types of atherosclerotic lesions. Antigen and activity levels of PAI-1 were determined in protein extracts of intimal and medial layers. In situ hybridization and immunohistochemical analyses were performed on serial sections of representative tissue specimens. RESULTS: Total PAI-1 antigen consistently increased from macroscopically normal areas (MNAs) to early lesions (ELs) and to maximal levels in fibrous (FPs) and calcified (CPs) plaques. No PAI activity was detected, although PAI-1 in its free form was present in all vascular specimens. Both free PAI-1 and PAI-1 complexed with plasminogen activators were significantly increased in extracts of advanced lesions. However, there was a 2-3 fold molar excess of free versus complexed PAI 1 in FPs and CPs. These findings suggest the presence of relevant amounts of PAI 1 in its substrate rather than in its inhibitor conformation in areas of advanced lesions. Compared with MNAs, PAI-1 mRNA was strongly expressed within the thickened intima of ELs. The highest PAI-1 expression was observed in FPs and CPs, being mainly localized in areas surrounding the necrotic cores in co localization with infiltrating macrophages. CONCLUSIONS: PAI-1 content is consistently increased in relation to the severity of the lesions in atherosclerotic coronary arteries. The concomitant elevation of PAI-1 mRNA suggests that the PAI-1 increase in regulated by local synthesis in the areas of atherosclerotic lesions. PMID- 9415270 TI - Soluble E-selectin, ICAM-1 and VCAM-1 levels in systemic and coronary circulation in patients with variant angina. AB - OBJECTIVE: The purpose of this study was to assess whether the plasma levels of soluble adhesion molecules including E-selectin, intercellular adhesion molecule 1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) are elevated in patients with variant angina and whether they are released in the coronary circulation. METHODS: Antecubital venous plasma samples were collected from 33 patients with variant angina, 22 patients with stable effort angina and 20 control subjects. Samples were also collected from the aortic root (AO) and the coronary sinus (CS) in 18 patients with variant angina before and after left coronary spasm induced by intracoronary injection of acetylcholine. The soluble adhesion molecules were assayed by enzyme immunoassay. RESULTS: Antecubital venous plasma soluble E-selectin (P < .05), ICAM-1 (P < .01) and VCAM-1 (NS) levels were higher in the variant angina group than in the control group, respectively. The plasma soluble ICAM-1 level was also higher (P < .01) in the variant angina group than in the stable effort angina group. In the variant angina group, both soluble ICAM-1 (P < .05) and VCAM-1 (P < .01) levels were significantly lower in CS than AO at baseline. In contrast, after the spasm the plasma soluble ICAM-1 level was (P < .05) higher in CS than AO and the CS-AO differences of soluble ICAM-1 (P < .05) and VCAM-1 (P < .05) increased as compared with the baseline, respectively. These values were remained unchanged in the stable effort angina group after rapid atrial pacing and in the control group after administration of acetylcholine. CONCLUSIONS: Circulating plasma levels of both soluble E-selectin and ICAM-1 were elevated in patients with variant angina, indicating an association of an inflammatory reaction with coronary spasm. Both soluble ICAM-1 and VCAM-1 appeared to be trapped in the coronary circulation at baseline and released into the coronary circulation following coronary spasm and reperfusion in the patients. PMID- 9415271 TI - DNA content, ploidy level and number of nuclei in the human heart after myocardial infarction. AB - OBJECTIVE: Cardiac hypertrophy due to a prolonged functional activity is associated with an increase of cell size and polyploidization of the myocyte nuclei. Myocardial infarction is characterized by loss of myocytes. Increased load and as a consequence hypertrophic growth of the surviving myocardium has to be expected. The aim of this study was to investigate the response of cardiomyocytes after infarction. METHOD: Biochemical and cytophotometric analysis was performed on myocyte and connective tissue nuclei to determine whether the human heart after myocardial infarction is accompanied by an increase in the ploidy level, DNA content and in the number of nuclei. A total of 15 hearts obtained from autopsy material was studied, among them 8 after myocardial infarction. The number of nuclei was measured by indirect computation. RESULTS: We found a decrease of 4c and no significant difference of 2c nuclei in infarcted hearts. DNA ploidy level (> 8c) as well as the proportion of aneuploid myocyte nuclei were increased in infarcted hearts. DNA concentration and total DNA content were increased in the hearts after myocardial infarction. Numerical ratio of connective tissue nuclei/myocyte nuclei, total number of nuclei, number of myocyte nuclei and number of connective tissue nuclei were increased in infarcted hearts. CONCLUSION: Polyploidization and nuclear hyperplasia of myocytes may represent an adaptive response of the myocardium to an ischemic injury. PMID- 9415272 TI - Evidence for a role for both the adenosine A1 and A3 receptors in protection of isolated human atrial muscle against simulated ischaemia. AB - OBJECTIVE: Adenosine receptor activation has been implicated in the mechanism of ischaemic preconditioning protection. Evidence suggests adenosine A1 receptor involvement, and possibly A3 receptor involvement in the rabbit. This study investigated the roles of these receptors in human preconditioning. Human A1- and A3-selective compounds were chosen based on Ki values for inhibition of N6-(4 amino-3-[125I]iodobenzyl)adenosine (125I-ABA) binding to stably expressed recombinant human A1 and A3 receptors. Cyclopentyladenosine (CPA), a 194-fold selective A1 agonist, and iodobenzylmethylcarboxamidoadenosine (IBMECA), a 10 fold selective A3 agonist were used alone and in combination with dipropylcyclopentylxanthine (DPCPX) a 62-fold selective A1 antagonist. METHODS: Human atrial trabeculae were superfused with oxygenated Tyrode's solution. After stabilisation, muscles underwent one of 8 protocols (n = 6 per group), followed by 90 min of simulated ischaemia and 120 min of reoxygenation. The experimental endpoint was recovery of contractile function, presented as percentage baseline function. RESULTS: 5 nM CPA (52.2 +/- 3.1%), 30 nM IBMECA (49.7 +/- 3.8%) and preconditioning (55.3 +/- 2.5%) produced similar functional recoveries at 120 min of reoxygenation; significantly different to controls (27.7 +/- 1.0%; P < 0.05, ANOVA). When DPCPX (200 nM) was added prior to 5 nM CPA, protection was lost (31.8 +/- 0.9%), but when added prior to 30 nM IBMECA, muscles continued to be significantly protected (41.5 +/- 2.3%). CONCLUSIONS: In human atrium both A1 and A3 receptor stimulation appears to mimic ischaemic preconditioning. This may represent the first evidence for A3 receptor involvement in 'pharmacological' preconditioning of human myocardium. PMID- 9415273 TI - Role of endothelin, nitric oxide and L-arginine release in ischaemia/reperfusion injury of rat heart. AB - OBJECTIVES: We tested the hypothesis that endothelin-1 (ET-1) aggravates ischaemia/reperfusion injury by stimulating cellular L-arginine depletion, which would result in reduced synthesis of nitric oxide (NO) and withdrawal of cardioprotection. METHODS: Five groups of rat hearts (n = 5 each) were perfused at 9 ml/min per g for 45 min, subjected to 15 min total global ischaemia and reperfused for 30 min; they received, from 5 min pre-ischaemia to end of reperfusion, either vehicle, L-arginine (1 mmol/l), the NO donor S-nitroso-N acetyl-DL-penicillamine (SNAP; 200 mumol/l), the inhibitor of NO formation NG nitro-L-arginine (L-NNA; 200 mumol/l), or the ET receptor antagonist PD 142893 (200 nmol/l). Cardiac function and release of L-arginine, cyclic GMP and lactate dehydrogenase (LDH) into coronary effluent were measured. RESULTS: Systolic, diastolic, and coronary reperfusion function were consistently improved by L arginine, SNAP, or PD 142893, but worsened by L-NNA (P < 0.05 in each case). L arginine release was transiently increased up to 25-fold on reperfusion (vehicle); release was reduced by SNAP (mean: 68%) and entirely prevented by PD 142893. Despite the increased outflow of L-arginine, formation of cyclic GMP was not reduced, but enhanced in reperfusion (11-fold; vehicle), and SNAP further augmented this release, but L-NNA had no significant effect. Release of LDH was decreased by L-arginine, SNAP, and PD 142893 in reperfusion. Finally, release of ET-1 was inhibited by NO in normoxia as well as throughout reperfusion as evident from the stimulatory effect of L-NNA. CONCLUSION: In ischaemia, ET-1 cause cell necrosis and L-arginine outflow without compromising NO synthesis in this model. PMID- 9415274 TI - Membrane phosphorylation protects the cardiac sarcoplasmic reticulum Ca(2+) ATPase against chlorinated oxidants in vitro. AB - OBJECTIVE: The calcium (Ca) pump of cardiac sarcoplasmic reticulum (SR) membranes is vulnerable to oxidation and hence likely to be damaged by chlorinated compounds, specifically hypochlorite (NaOCl) and monochloramine (NH2Cl), the most potent oxidants produced upon neutrophil activation. This could occur during prolonged ischemia or myocardial infarction when tissue levels of catecholamines are high. Phospholamban (PLN), the phosphorylatable regulator of the Ca pump, plays a central role in the effects of beta-adrenergic agonists on the heart. The purpose of this study was to investigate a possible role of PLN in determining the pump's sensitivity to NaOCl and NH2Cl. METHODS: Ca-uptake and Ca(2+)-ATPase activities in purified phosphorylated and control canine cardiac microsomes, incubated at increasing concentrations of NaOCl or NH2Cl, were related to the extent of PLN phosphorylation by protein kinase A, which was quantitated by PhosphorImager analysis. RESULTS AND CONCLUSIONS: Our data indicate that microsomal phosphorylation protects the Ca pump fully against 10 microM NaOCl or NH2Cl, which inhibit Ca-uptake by 21-41% when assayed at 25 or 37 degrees C and saturating Ca2+ in unphosphorylated microsomes, and protects partially at higher oxidant concentrations. The protective effect of protein kinase A on Ca-uptake is proportional to the amount of phosphorylated PLN. No comparable protection against similar oxidative damage of the Ca pump is observed when light fast skeletal muscle microsomes, which lack PLN, are incubated under conditions favorable for phosphorylation nor when PLN's inhibition of the cardiac Ca pump is relieved by proteolytic cleavage of its cytoplasmic domain. Our findings contribute toward an understanding of possible endogenous protective mechanisms that may promote calcium homeostasis in myocardial cells in inflammatory states associated with neutrophil activation and may suggest an approach toward development of protective strategies against oxidative damage in the heart. PMID- 9415275 TI - Pharmacodynamics of basic fibroblast growth factor: route of administration determines myocardial and systemic distribution. AB - OBJECTIVE: We have shown that basic fibroblast growth factor (bFGF/FGF-2) enhances myocardial collateral development in a canine model of progressive coronary occlusion when delivered via the left atrial or intracoronary routes; however, we have found intravenous bFGF ineffective in the same model. Data on the fate and efficacy of intravenous bFGF are limited. We hypothesized that first pass lung uptake might limit myocardial bFGF availability after intravenous injection. We postulated that delivery of bFGF through the distal port of a wedged Swan Ganz catheter might circumvent this problem by restricting exposure of bFGF to a limited number of pulmonary binding sites. This study evaluated differential regional uptake of 125I labeled bFGF following bolus intravenous, Swan Ganz, left atrial, intracoronary, and pericardial delivery. METHODS: Mongrel dogs were used. Human recombinant bFGF, monoiodinated with 125I, was mixed with cold bFGF to a specific activity of 0.03 microCi/microgram. Approximately 100 micrograms/kg was injected per animal by the intravenous, left atrial, Swan Ganz, intracoronary, or pericardial route. Dogs were killed 15 min or 150 min later. The heart, lungs, liver, spleen, and kidneys were harvested and 125I activity was assessed. Immunohistochemical and pharmacokinetic studies were also performed. RESULTS: Serum half life of bFGF was comparable after intracoronary, intravenous and left atrial delivery (50 min); however, there were significant differences with regard to pharmacodynamics. After intracoronary administration, 3-5% of the total bFGF dose was recovered from the heart, with the peptide immunolocalized to the extracellular matrix and vascular endothelium. In contrast, only 1.3% of the injected bFGF was localized to the heart after left atrial administration and 0.5% was recovered after intravenous or Swan Ganz delivery. Pericardial administration resulted in substantial cardiac bFGF delivery; 19% was present at 150 min. Myocardial uptake was similar with Swan Ganz and intravenous delivery, suggesting that the administered dose did not saturate available pulmonary binding sites. CONCLUSIONS: These data predict efficacy of intracoronary, left atrial, and pericardial bFGF for myocardial angiogenesis, and a lack of efficacy after bolus intravenous and Swan Ganz administration. PMID- 9415276 TI - Increased Ca2+ sensitivity of contractile elements via protein kinase C in alpha toxin permeabilized SMA from young spontaneously hypertensive rats. AB - OBJECTIVE: The purpose of the present investigation was to examine the Ca2+ sensitivity of the contractile elements via protein kinase C (PKC) in superior mesenteric artery (SMA) from young (5-6 weeks old) spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto rats (WKY). METHODS: Staphylococcal aureus alpha-toxin, which produces pores in the plasma membrane too small to allow passage of proteins such as PKC, was used to investigate the signal transduction system in vascular smooth muscle cells. We investigated the Ca2+ sensitivity of the contractile apparatus via PKC in intact and alpha-toxin skinned SMA from young SHR and WKY. RESULTS: In intact SMA, high K+ responses were not different between SHR and WKY. However, phorbol 12,13-dibutyrate (PDBu, a PKC activator) augmented high K(+)-evoked contractions and PKC inhibitors, such as 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7) and calphostin C, suppressed them more in SHR as compared with WKY. In alpha-toxin skinned SMA, the [Ca2+]i-force relationship curve was not significantly different between SHR and WKY. However, PDBu augmented [Ca2+]i-evoked contractions and PKC inhibitors suppressed them more in SHR than in WKY. CONCLUSION: These results suggest that the Ca2+ sensitivity of the contractile elements via PKC is significantly greater in prehypertensive SHR than in age-matched WKY. This abnormality in small muscular arteries may be involved in the pathogenesis of hypertension in SHR. PMID- 9415277 TI - Low density lipoprotein enhances the thrombin-induced growth of vascular smooth muscle cells. AB - OBJECTIVE: In the present study we investigated whether low density lipoprotein is able to enhance the growth promoting effects of thrombin in vascular smooth muscle cells. METHODS: DNA synthesis was examined by measurement of the [3H]thymidine incorporation into the cell DNA. Cell count was measured with a Neubauer cell box. Thrombin receptor mRNA was determined by Northern blotting. Ca2+ was measured by the fura 2-method. RESULTS: Thrombin (5 nmol/l), thrombin receptor activating protein (3 mumol/l) and low density lipoprotein (33 nmol/l) induce a 652 +/- 80%, 593 +/- 80% and a 316 +/- 60% increase in [3H]thymidine incorporation into DNA (mean +/- SD, n = 3), respectively. A coincubation of thrombin or thrombin receptor activating protein with low density lipoprotein led to a 1245 +/- 160% or 1200 +/- 40% increase of DNA synthesis (mean +/- SD, n = 3). Thus, coincubation of low density lipoprotein and thrombin causes a synergistic rather than an additive mitogenic effect on smooth muscle cells. Thrombin and low density lipoprotein induced a 22 +/- 8.4% and a 29% +/- 6% increase in cell number, respectively. Simultaneous treatment of vascular smooth muscle cells with thrombin and low density lipoprotein caused a 63 +/- 14% increase in cell number (mean +/- SD, n = 3). To further elucidate the underlying mechanism, we studied the effect of low density lipoprotein on the expression of thrombin receptor mRNA. Low density lipoprotein caused a 2.5-fold increase of thrombin receptor mRNA within 24 h, as assessed by Northern analysis. Preincubation of cells for 24 h with 33 nmol/l low density lipoprotein resulted in an elevation of the thrombin-induced increase in cytosolic free Ca2+ concentration from 538 +/- 54 to 923 +/- 75 nmol/l (mean +/- SD, n = 4). CONCLUSION: In summary, low density lipoprotein may enhance the mitogenic effect of thrombin probably by an up-regulation of thrombin receptor gene expression in vascular smooth muscle cells or by an elevation of the thrombin-induced increase in cytosolic free Ca2+ concentration. PMID- 9415278 TI - Differences in development of coronary arteries and veins. AB - OBJECTIVE: The differentiation of the coronary vasculature was studied to establish in particular the formation of the coronary venous system. METHODS: Antibody markers were used to demonstrate endothelial, smooth muscle, and fibroblastic cells in serial sections of embryonic quail hearts. The anti-beta myosin heavy chain and the neuronal marker HNK-1 were added to our incubation protocol. RESULTS: In HH32, the coronary vascular network has developed into a circulatory system with connections to the sinus venosus, the aorta and the right atrium. The connections between the aorta and the right atrium allow for direct arteriovenous shunting. Subsequently, differentiation into coronary arteries and veins occurs with an interposed capillary network. The smooth muscle cells of the coronary arterial media derive from the subepicardial layer, whereas the subepicardially located cardiac veins recrute atrial myocardium, as these cells express the beta-myosin heavy chain antigen. Ganglia are located in the subepicardium close to the vessels, while nerve fibres tend to colocalize with the formed vessel channels. CONCLUSIONS: A new finding is presented in which the subepicardial coronary veins have a media that consists of myocardial cells. The close positional relationship of neural tissue and coronary vessels that penetrate the heart wall is explained as inductive for vessel wall differentiation, but not for invasion into the heart. PMID- 9415279 TI - Effects of sympathetic nerve blockade on vasoconstrictive properties of nitric oxide synthase inhibition in sheep. AB - OBJECTIVE: Inhibition of nitric oxide synthase causes intense vasoconstriction. This effect is thought to be dependent on sympathetic nerve activity. Thus, we investigated the vasoconstrictive effects of the nitric oxide synthase inhibitor NG-nitro-L-arginine methyl ester (L-NAME) in sheep, in which a reversible sympathetic block was established by thoracic epidural anesthesia. METHODS: Sheep (n = 11) were surgically prepared for chronic study. After at least 5 days of recovery, L-NAME was continuously administered and hemodynamics were monitored. This was done in sheep with and without sympathetic blockade in randomized order. RESULTS: The vasoconstrictive effects of L-NAME were similar in sheep with and without sympathetic blockade. CONCLUSION: The obtained results suggest that the vasoconstrictive properties of nitric oxide synthase inhibitors are independent of sympathetic tone. PMID- 9415280 TI - The A10 cell line: a model for neonatal, neointimal, or differentiated vascular smooth muscle cells? AB - OBJECTIVES: The A10 cell line was derived from the thoracic aorta of embryonic rat and is a commonly used model of vascular smooth muscle cells (VSMC). Despite its wide use this cell line has not been well characterized. This is especially important in light of recent evidence of phenotypically distinct cell populations isolated from rat vascular tissue. Therefore, the present study was undertaken to confirm the VSMC nature of A10 cells and to investigate whether these cells particularly resemble adult, neonatal, or neointimal rat VSMC. METHODS: A variety of defining characteristics were used that included immunofluorescent analysis for smooth muscle alpha-actin, smooth and non-muscle myosin heavy chains, desmin and vimentin; Western analysis for smooth muscle and non-muscle myosin heavy chains; mRNA analysis for smooth muscle myosin heavy chain, calponin, SM22 alpha, tropoelastin and PDGF-B peptide; and functional assays of cell migration, proliferation and agonist induced intracellular Ca transients. RESULTS: A10 cells expressed smooth muscle alpha-actin, SM22 alpha, smooth muscle calponin and vimentin, characteristic of in vivo rat VSMCs; however they also resembled de differentiated smooth muscle cells in that they expressed non-muscle myosin rather than smooth muscle myosin heavy chain. A10 cells resembled cultured rat neonatal smooth muscle cells ("pup cells") in that they had an epithelioid shape and lacked functional PDGF-alpha receptors: however they did not express PDGF-B mRNA or proliferate in low serum containing medium as do neonatal cells. A10 cells had several characteristics in common with neointimal cells including the expression of alpha-actin, vimentin, and non-muscle myosin and the lack of expression of PDGF-B mRNA as well as the ability to migrate in response to PDGF BB. CONCLUSION: In conclusion, A10 cells are nondifferentiated VSMC that differ from neonatal but bear significant resemblance to neointimal cells. PMID- 9415281 TI - Are M cells present in the ventricular myocardium of the pig? A question of maturity. PMID- 9415282 TI - Gene therapy in the cardiovascular system. PMID- 9415283 TI - Optimal techniques for arterial gene transfer. AB - Cardiovascular gene therapy is becoming a clinical reality due to improved vectors, delivery systems and careful experimental validation studies. Nearly all cardiovascular diseases are amenable to gene therapy, but the optimal combination of vector, delivery system and therapeutic gene is likely to be unique to each application. Currently, the most efficient vectors available are replication defective adenoviral vectors, but transgene expression is limited in time due to a strong immune response. Conversely, non-viral vectors or plasmid DNA may be used safely but have very limited efficiency. Percutaneous, catheter-based delivery is feasible for most applications. The ultimate issues that will decide of the future of gene therapy are safety of the transfer and delivery techniques as well as cost/effectiveness comparisons with alternative therapies, including local delivery of drugs, proteins and/or mechanical devices. PMID- 9415284 TI - Local drug delivery systems and prevention of restenosis. PMID- 9415285 TI - The use of mRNA differential display for discovery of novel therapeutic targets in cardiovascular disease. AB - Recent advances in molecular biology techniques have provided powerful tools for discovery of novel genes relevant to both biological and pathological processes. mRNA differential display is an emerging technique for novel gene discovery and it has been successfully applied to many physiological and pathological conditions including normal development, cell differentiation, cancer, cardiovascular disease, inflammation and CNS disorders. In the present work, we briefly illustrate the critical procedure and highlight most recent technical improvements and modifications of this technology. Based upon the successful applications of this technique in cardiovascular research, it may provide a valuable and powerful investigational tool for the identification of novel therapeutic targets in cardiovascular diseases. PMID- 9415286 TI - Prospects for adenovirus-mediated gene therapy of inherited diseases of the myocardium. PMID- 9415287 TI - Gene transfer and cell transplant: an experimental approach to repair a 'broken heart'. PMID- 9415288 TI - Prevention of vein graft failure: potential applications for gene therapy. AB - The use of gene therapy in the clinical setting is believed to be a realistic option for the future. Many clinical trials are underway for treatment of disorders as diverse as cancer, peripheral vascular disease, and numerous monogenic disease. However, gene therapy for vein graft failure may be more distant due to the highly complex, multifactorial aetiology of the disease. Although many of the cellular mechanisms involved in vein graft failure have been reported, important barriers still need to be overcome before gene therapy could become a clinical reality. Further understanding of the molecular mechanisms involved in graft failure will lead to the identification of appropriate therapeutic genes. Moreover, limitations in the current delivery systems need to be overcome to allow efficient, safe delivery and expression of transgenes for the required length of time in vivo. However, currently available gene delivery vectors are extremely useful tools to help in our understanding of vein graft failure. In this review, we address the issues surrounding gene therapy with particular emphasis on its future potential to ameliorate long term vein graft occlusion. PMID- 9415289 TI - Adenovirus gene therapy for hypercholesterolemia, thrombosis and restenosis. PMID- 9415290 TI - Gene therapy for arterial thrombosis. AB - Conventional antithrombotic treatments with antiplatelet, anticoagulant or fibrinolytic drugs are not uniformly successful and are associated with hemorrhagic side effects. Thus, new approaches to the prevention and treatment of arterial thrombosis are desirable. The gene transfer approach is particularly attractive because of its unique ability to express an antithrombotic gene at selected sites of the vessel wall (where thrombosis is threatened) while avoiding systemic anticoagulation. Clinical conditions potentially amenable to antithrombotic gene therapy include coronary artery bypass grafting, percutaneous transluminal coronary angioplasty, peripheral artery angioplasty or thrombectomy, intravascular stenting, and vascular graft prostheses. Gene therapy may prove effective in preventing subacute thrombosis in these settings and, eventually, may play an adjuvant role to systemic thrombolysis in the treatment of acute arterial occlusion. The introduction of an antithrombotic gene into the arterial wall can be achieved either by direct in vivo gene transfer (e.g., by luminal administration of a viral vector) or by in vitro genetic manipulation of cells before their seeding onto vascular grafts, stents, or denuded arteries. The direct gene transfer approach has been used to deliver antithrombotic genes to animal arteries in vivo. Antithrombotic genes used to date include those encoding enzymes of the prostacyclin synthetic pathway, nitric oxide synthase, the thrombin inhibitor hirudin, and thrombomodulin. The in vitro gene transfer approach has been used to enhance the fibrinolytic activity of vascular grafts by overexpressing plasminogen activators. If the initial successes of gene therapy for thrombotic disease in animal models are confirmed by longer-term experiments, and if new vectors are developed which permit prolonged transgene expression without inflammation, human studies can be initiated. PMID- 9415291 TI - Arterial gene transfer of acidic fibroblast growth factor for therapeutic angiogenesis in vivo: critical role of secretion signal in use of naked DNA. AB - OBJECTIVE: Previous studies have demonstrated that arterial gene transfer of naked DNA encoding for a secreted protein may permit modulation of the host phenotype despite a low transfection efficiency. Acidic fibroblast growth factor (aFGF) is an angiogenic growth factor, but is not secreted by intact cells. In the current study, we investigated the hypothesis that addition of a hydrophobic leader sequence to achieve active secretion of the gene product would permit therapeutic angiogenesis following arterial gene transfer of naked DNA encoding for aFGF. METHODS: Ten days following surgical induction of unilateral hindlimb ischemia, New Zealand white rabbits were randomized to intra-arterial gene transfer with one of three plasmids: p267 (encoding non-secreted aFGF, n = 10), pMJ35 (encoding secreted aFGF) (n = 10), or 500 micrograms of pGSVLacZ (control, n = 10) (500 micrograms each). All animals were studied at 30 days post-gene transfer for evidence of therapeutic angiogenesis. RESULTS: pMJ35 transfectants had more angiographically visible collaterals (angiographic score = 0.76 +/- 0.02) than either p267 (0.55 +/- 0.02, p < 0.01) or LacZ (0.47 +/- 0.02, p < 0.001). Limb blood pressure ratio for pMJ35 was 0.88 +/- 0.02 vs. 0.68 +/- 0.04 for p267 (p < 0.01) and 0.57 +/- 0.04 for LacZ (p < 0.001). Vascular resistance was significantly lower in the pMJ35 group, compared with that in pGSVLacZ group, both in resting state (3.2 +/- 0.4 vs. 7.4 +/- 1.4 respectively, p < 0.05) and after the administration of nitroprusside. Capillary density (per mm2) was also superior in pMJ35 group (274 +/- 10) vs. p267 (204 +/- 9, p < 0.01) and LacZ (177 +/- 6, p < 0.001). CONCLUSION: The paracrine effects of a secreted gene product may obviate the need for adjunctive vectors in strategies of arterial gene therapy. PMID- 9415292 TI - Gene therapy for collateral vessel development. AB - Transfer of the cDNA coding for angiogenic factors represents a novel and promising approach to induce therapeutic angiogenesis and enhance blood flow to ischemic tissues which cannot be revascularized otherwise. This review will focus on therapeutic angiogenesis based on gene transfer techniques for the treatment of myocardial and limb ischemia. The experimental studies demonstrating the angiogenic effect of recombinant growth factors in animal models and in humans, as well as the most promising methods for gene transfer, will be described. Further, gene transfer studies to induce therapeutic angiogenesis will be reviewed to identify critical questions that still need to be answered before gene therapy with angiogenic factors may be considered for routine clinical application. PMID- 9415294 TI - Vectors based on Semliki Forest virus for rapid and efficient gene transfer into non-endothelial cardiovascular cells: comparison to adenovirus. AB - OBJECTIVE: Replication-deficient, recombinant adenovirus is used as a carrier for gene transfer, but it is unspecific and the onset of transgene expression is relatively late. Here, we evaluated the efficiency and selectivity of gene transfer mediated by recombinant Semliki Forest virus (SFV). METHODS: We compared the efficiency of a SFV-based vector with an adenoviral vector, using LacZ as a reporter gene. Firstly, the affinity for vascular smooth muscle cells, endothelial cells and cardiac myocytes was assessed. Secondly, we compared the time course of LacZ expression and cytotoxicity in vascular smooth muscle cells. RESULTS: The SFV-based vector infects vascular smooth muscle cells and cardiomyocytes as efficiently as adenovirus. In contrast to adenovirus, SFV hardly transfers LacZ to endothelial cells (2.6% or less). SFV-mediated expression was visible after 1 h, reaching a maximum after 6 h. In contrast, adenovirus-mediated expression became visible after 6 h, and reached a maximum after 48-72 h. Both vectors were cytotoxic. CONCLUSIONS: We demonstrate that SFV efficiently transfers LacZ to vascular smooth muscle cells and cardiomyocytes, but not to endothelial cells. In contrast, adenovirus causes efficient transgene expression in all cell types tested. Furthermore, SFV-mediated expression is faster than adenovirus-mediated expression. Therefore, SFV-mediated gene transfer may be a suitable alternative to adenovirus, providing a fast expression in non endothelial cardiovascular cell types. PMID- 9415293 TI - Angiogenesis by gene therapy: a new horizon for myocardial revascularization? AB - The concept of therapeutic angiogenesis is based on the premise that the potential for vascular growth inherent in vascular tissue can be utilized to promote the development of new blood vessels under the influence of the appropriate growth factors. Direct application of growth factors of the fibroblast (acidic, basic fibroblast growth factor, FGF-5), endothelial (vascular endothelial growth factor) and other series has been effective in preliminary studies. Angiogenesis by gene transfer provides an attractive alternative, with the advantage that the protein may continue to be secreted for a longer period of time and that the gene may be targeted to specific tissues to enhance efficacy and reduce systemic side effects. Angiogenesis by gene transfer is currently under investigation using a variety of growth factors and a wide array of potential delivery systems. These include application of the gene as naked DNA or by viral vector in the proximal vessel by direct intravascular injection, interventional cardiologic techniques (hydrogel coating on balloon, double balloon system, stent implantation) or by direct application to adventitia, pericardium or ischemic tissue distal to the site of arterial obstruction. As our understanding of the molecular and genetic processes underlying angiogenesis increases, and as we examine the results of preliminary animal and human protocols, we hope to develop the potential of angiogenesis by gene transfer for therapeutic use. PMID- 9415295 TI - Efficient adenoviral gene transfer to early venous bypass grafts: comparison with native vessels. AB - OBJECTIVES: Gene therapy may provide new approaches to reduce vein graft failure following coronary or peripheral bypass surgery. The aim of this study was to investigate the relative efficacy of intraoperative adenoviral gene transfer to vein grafts, comparing transgene expression in vein grafts with that in matched native vessels in the same animal. In addition, we assessed the impact of bypass grafting on the cellular targets of gene transfer. METHODS: New Zealand White rabbits underwent interposition bypass grafting of the carotid artery, using the ipsilateral external jugular vein, which was infected with an adenovirus expressing beta-galactosidase immediately prior to bypass grafting (n = 16). The contralateral native jugular vein (n = 16) and carotid artery (n = 8) were infected concurrently with the same adenoviral preparation. After 3, 7 or 14 days, beta-galactosidase protein expression was quantified by ELISA, and specific cell types expressing beta-galactosidase were identified by X-Gal staining and by immunohistochemistry. RESULTS: After 3 days, endothelial cells were efficiently transduced in all vessels; medial smooth muscle cells were transduced infrequently. In contrast to jugular veins after gene transfer, endothelium in vein grafts showed expression of VCAM-1 and ICAM-1, and intense inflammation with CD18+ leukocytes. Transgene expression in vein grafts at day 3 was maintained at levels approximately 50% of that in ungrafted jugular veins, but continued to decrease through day 7. CONCLUSIONS: Although vascular injury in early venous bypass grafts reduces gene transfer efficacy, significant transgene expression is maintained for at least 7 days. These findings have important implications for intraoperative gene transfer strategies in vein grafts. PMID- 9415296 TI - Gene transfer into vascular cells using adeno-associated virus (AAV) vectors. AB - OBJECTIVES: Recombinant viral vectors based on the nonpathogenic parvovirus, adeno-associated virus (AAV), have a number of attractive features for gene therapy, including the ability to transduce non-dividing cells and its long-term transgene expression. In this study, an AAV vector containing bacterial beta galactosidase gene (lacZ) was used to transduce cultured rat vascular smooth muscle cells (VSMC) in vitro and rat thoracic aortas ex vivo. METHODS: VSMC were transduced with AAV-lacZ at multiplicities of infection (MOI) ranging from 5.0 x 10(5) to 1.0 x 10(7). Expression of beta-galactosidase (beta-gal) in VSMC was evaluated by X-gal staining and a beta-gal ELISA method. Excised rat aortas were incubated with medium containing AAV-lacZ. Expression of beta-gal in the aortic segments was evaluated by X-gal staining. RESULTS: With increasing MOI, up to 50% of cultured VSMC were positive by X-gal staining and the beta-gal expression increased up to 15 ng/mg protein. The expression gradually decreased during the culture but was detectable for at least 1 month. In the ex vivo study, AAV vectors transduced endothelial and adventitial cells in rat aortic segments, while no expression was seen in medial VSMC. CONCLUSIONS: AAV vectors can efficiently transduce rat VSMC in vitro. AAV-mediated ex vivo gene transfer into the normal aorta resulted in efficient gene transfer into endothelial and adventitial cells but not into medial VSMC. These findings suggest that AAV-based vectors are promising for use in cardiovascular gene therapy. PMID- 9415297 TI - Transfection of human endothelial cells. AB - OBJECTIVE: The introduction of recombinant genes into endothelial cells provides a method to study specific gene products and their effect on cell function. In addition, endothelial cells can be used for implantation into vessels or prosthetic vascular grafts. Because transfection efficiencies in human endothelial cells have been low, it is important to develop improved gene transfer techniques. Therefore, several transfection methods were optimized and transfection efficiencies were determined. METHODS: Transfection by particle mediated gene transfer (biolistics) or by cationic liposomes were optimized and compared to calcium phosphate and DEAE-dextran. Transfection efficiency was determined using either a beta-galactosidase or placental alkaline phosphatase reporter gene. The effect of promoter strength was analyzed by transfecting plasmids with either the Rous sarcoma virus (RSV) promoter or cytomegalovirus (CMV) promoter regions. RESULTS: Optimal conditions for particle-mediated gene transfer utilized gold particles of 1.6 microns diameter, a target distance of 3 cm, helium pressures of 8.96 MPa (1300 psi) and cell confluence of 75%. Transfection with different cationic liposomes demonstrated that one compound, N (3-aminopropyl)-N,N-dimethyl-2,3-(bis-dodecyloxy)-1-propanimi nium bromide/dioleoyl phosphatidylethanolamine (gamma AP-DLRIE/DOPE), was optimal for gene transfer when 5 micrograms of DNA and 10 to 20 micrograms of lipid was used. With both gold particles and gamma AP-DLRIE/DOPE, the alkaline phosphatase reporter was more efficient than beta-galactosidase using comparable promoters and polyadenylation sites. CMV regulatory elements were more efficient than the RSV promoter in optimizing gene expression. Optimal gene transfer efficiency was 20.28% of cells with gamma AP-DLRIE/DOPE, 3.96% with biolistics, 2.09% with calcium phosphate and 0.88% with DEAE-dextran. CONCLUSIONS: Gene expression is detectable in a high percentage of human endothelial cells after liposome mediated transfection when expression is controlled by a strong promoter. Particle-mediated transfection is less efficient under these conditions, but more effective than liposomes when expression is driven by a relatively weak promoter. Calcium phosphate and DEAE-dextran are less useful. PMID- 9415298 TI - Immunosuppression prolongs adenoviral mediated transgene expression in cardiac allograft transplantation. AB - BACKGROUND: The immune response to adenoviral vectors used in gene transfer limits the duration of transgene expression and thus poses a potential limitation to their effectiveness for gene therapy. The need for immunosuppression in transplantation may modify this immune response and facilitate prolonged transgene expression. This study hypothesizes that in the setting of heart transplantation, the use of routine immunosuppression will prolong adenoviral mediated transgene expression. METHODS AND RESULTS: In a model of rat heterotopic abdominal heart transplantation, 350 microliters of viral solution (1 x 10(9) pfu/ml) was infused into the coronary arteries of donor hearts at the time of procurement. The duration of transgene expression was examined following (a) syngeneic transplantation in non-immunosuppressed animals (group A): (b) syngeneic transplantation in immunosuppressed animals (group B); and (c) allogeneic transplantation in immunosuppressed animals (group C). After transplantation donor hearts were studied at; 1, 4, 8 and 12 weeks. Transgene expression was assessed by histochemical staining of tissue cross sections for beta-galactosidase activity. In the non-immunosuppressed syngeneic group (group A), transgene expression had largely disappeared by 4 weeks, whereas in both the immunosuppressed syngeneic (group B) and immunosuppressed allogeneic (group C) animals expression of the reporter gene persisted for the 12 weeks of the study, although the level of expression decreased significantly over time. CONCLUSIONS: This study demonstrates that transgene expression using adenoviral vectors is prolonged by immunosuppression in the heart transplantation setting. PMID- 9415299 TI - Percutaneous delivery of the gax gene inhibits vessel stenosis in a rabbit model of balloon angioplasty. AB - OBJECTIVES: The expression of gax, an anti-proliferative homeobox gene, is rapidly downregulated in vascular smooth muscle cells (VSMCs) following arterial injury. Here we performed percutaneous adenovirus-mediated gene transfer into the iliac arteries of normal rabbits using a channel balloon catheter to assess the effects of gax overexpression on neointima formation, lumen diameter, reendothelialization and functional vasomotion. METHODS: A channel balloon catheter was used to perform both the arterial injury and local gene delivery. In each animal both iliac arteries were randomly assigned to receive either an adenovirus expressing the gax gene (Ad-Gax) or the beta-galactosidase gene (Ad beta gal). In a second group of animals arteries were randomly assigned to receive either Ad-beta gal or saline. RESULTS: At one month, angiography revealed 36% less luminal narrowing in the Ad-Gax-treated arteries relative to the Ad-beta gal-treated control arteries. Histological analysis revealed that the intimal/medial ratio (I/M) was reduced by 56% in the Ad-Gax group. Endothelium dependent vasomotion was not affected by the gax gene transfer. In the second group, no statistically significant differences were found between the saline and the Ad-beta gal-treated vessels for any of these parameters. CONCLUSIONS: Percutaneous adenovirus delivery of the gax gene to rabbit iliac arteries following endothelial denudation and vessel wall injury reduces neointimal hyperplasia and luminal stenosis, but does not affect endothelium-dependent vasomotion. This study demonstrates that a VSMC transcription factor can potentially be utilized for the development of a molecular therapy for vascular disorders. PMID- 9415300 TI - Microangiographic assessment of collateral vessel formation following direct gene transfer of vascular endothelial growth factor in rats. AB - OBJECTIVE: The development of collateral microvessels following therapeutic angiogenesis with vascular endothelial growth factor (VEGF) was investigated using a new system of microangiography that employs monochromatic synchrotron radiation (SR) and a high definition video system to visualize arteries with a spatial resolution of 30 microns. METHODS: Ischemia was induced in the hindlimb of 20 rats by excision of the femoral artery, followed by transfection of the plasmid (400 micrograms) encoding VEGF or beta-galactosidase (control) into limb muscles. Microangiography was used to assess the development of collaterals in the ischemic limb four weeks after treatment. RESULTS: Gene transfer of VEGF produced morphologically similar, but significantly more extensive, collateral networks at the microvascular level as compared with the naturally occurring collateral arteries in the control animals (angiographic score: 0.88 +/- 0.08 versus 0.54 +/- 0.05, p < 0.01). No adverse vascular effects such as hemangiomas and/or arteriovenous (AV) fistulae were observed following VEGF treatment. The vasodilator effect of papaverine was evident in relatively large vessels in both groups. At the microvascular level (diameter < 100 microns), however, papaverine induced significant vasodilation in the VEGF-treated animals, and almost no vasodilation in the controls. CONCLUSIONS: SR microangiography allowed us to assess the development of small collateral arteries following VEGF-gene transfer. The information obtained may provide new insights regarding the collateral microcirculation and therapeutic angiogenesis. PMID- 9415301 TI - Expression and function of a recombinant endothelial nitric oxide synthase gene in porcine coronary arteries. AB - OBJECTIVES: Direct gene transfer of exogenous nitric oxide synthase, with the subsequent increase in nitric oxide production, could represent a potential therapeutic strategy in the treatment of vascular proliferative disorders. The goal of the present study was to determine if porcine coronary arteries could be transduced with an adenoviral vector encoding endothelial nitric oxide synthase (Ad.CMVeNOS) resulting in functional expression. METHODS AND RESULTS: Segments of porcine right coronary artery were exposed for 1 h at 37 degrees C to either replication-deficient adenovirus encoding bovine endothelial nitric oxide synthase (Ad.CMVeNOS, 5 x 10(9) pfu/ml) or control adenovirus encoding Escherichia coli beta-galactosidase (Ad.CMVLacZ, 5 x 10(9) pfu/ml). Immunohistochemistry with a monoclonal antibody specific for nitric oxide synthase (NOS) demonstrated recombinant gene expression in both the endothelial and adventitial layers of Ad.CMVeNOS transduced coronaries with only endogenous NOS confirmed in the endothelium of Ad.CMVLacZ arteries. Coronary arteries transduced with Ad.CMVeNOS yielded 517 +/- 110 (mean +/- S.E.M.) nM/ng nitrite while vessels transduced with Ad.CMVLacZ yielded 126 +/- 84 nM/ng (P < 0.05, n = 6). Isometric tension recording, following prostaglandin F2 alpha constriction, documented a reduced tension in Ad.CMVeNOS transduced coronaries, compared to matched Ad.CMVLacZ coronaries (6.10 +/- 1.08 g vs. 8.45 +/- 1.19 g, respectively, P = 0.05, n = 8). This tension differential was eliminated with prior incubation in NG-monomethyl-L-arginine (L-NMMA, 10(-4) M). The EC50 for calcium ionophore relaxation of Ad.CMVeNOS coronary arteries was reduced compared to Ad.CMVLacZ ( 7.90 +/- 0.03 logM vs. -7.26 +/- 0.11 logM, respectively, P < 0.05, n = 8). CONCLUSIONS: These studies demonstrate successful transfer of endothelial nitric oxide synthase into porcine coronary arteries as verified by histochemical localization of recombinant protein with an increase of nitric oxide release as demonstrated by enhanced nitrite production and an alteration in vasomotor function. PMID- 9415302 TI - Analysis of tissue-specific gene delivery by recombinant adenoviruses containing cardiac-specific promoters. AB - OBJECTIVE: To approach heart muscle diseases by gene transfer, an adenoviral vector system was intended to be established suitable for gene expression in ventricular and/or atrial myocardium. METHODS: Two adenoviral vectors (Ad-mhcLuc, Ad-mlcLuc) were constructed, in which the luciferase reporter gene is under control of either the ventricle-specific myosin light chain-2 (mlc-2v) or the atrial- and ventricular-specific alpha-myosin heavy chain (alpha-mhc) promoter. For controls, a recombinant adenovirus without promoter (Ad-Luc) and one with the Rous sarcoma virus (rsv) promoter (Ad-rsvLuc) were generated. A volume of 20 microliters containing 2 x 10(9) plaque forming units (pfu) of the recombinant adenoviruses Ad-mhcLuc, Ad-mlcLuc, Ad-rsvLuc or Ad-Luc was injected into the cardiac cavity or the quadriceps femoris muscle of neonatal rats. After five days animals were sacrificed and nine different tissues were analyzed for reporter gene expression by detection of light activity relative to mg of tissue. RESULTS: Injections of recombinant adenoviruses into the cardiac cavity of neonatal rats resulted in heart-specific gene expression of Ad-mlcLuc (20 fold of Ad-Luc; 11% of Ad-rsvLuc), whereas Ad-mhcLuc gave mainly luciferase activity in the heart (6.5-fold of Ad-Luc; 3% of Ad-rsvLuc) with additional activity in lung and liver (2-4 fold of Ad-Luc). In the ventricular tissue Ad-mlcLuc revealed a 35-fold higher luciferase activity, whereas Ad-mhcLuc, Ad-rsvLuc and Ad-Luc showed only 2 fold higher luciferase activities compared to the atrium. Viral DNA in atrial and ventricular tissue was detected by PCR at approximately the same abundance independent of the injected type of adenovirus. Direct injection of Ad-mhcLuc and Ad-mlcLuc into the thigh muscle revealed only background luciferase activities. CONCLUSIONS: In the adenoviral system only the mlc-2v promoter may fulfil the safety requirements for a myocardial specific gene expression with a high selectivity for the ventricular myocardium, thus providing a promising tool for future gene therapy of cardiomyopathies. PMID- 9415303 TI - Regulated expression of a foreign gene targeted to the ischaemic myocardium. AB - OBJECTIVES: Regulated expression of transferred foreign genes may be an important feature of gene therapy. Because coronary artery disease often involves intermittent myocardial ischaemia followed by periods of normal cardiac function it will probably be necessary to regulate the expression of putative therapeutic/cardioprotective genes directly in response to ischaemia-associated signals. The objectives of the current study were to develop a combination of gene regulatory components that can be used to target a product to the myocardium and limit the expression of the gene to periods of ischaemic activity. METHODS: Expression plasmids were constructed containing muscle-specific promoters and hypoxia-responsive enhancer elements linked to a reporter gene. The regulation of these constructs by hypoxia or experimental ischaemia was measured following transient expression in cultured cells or after direct injection of DNA into the rabbit myocardium. RESULTS: A single set of hypoxia response elements placed immediately upstream of the minimal muscle-specific alpha-myosin heavy chain promoter conferred potent positive regulation of this promoter by hypoxia in vitro and by ischaemia in vivo. Induction by ischaemia persisted for at least 4 h and returned to the baseline level within 8 h. CONCLUSIONS: Hypoxia responsive regulatory elements, in combination with weak tissue-restricted promoters incorporated into an appropriate vector system may allow controlled expression of a therapeutic gene in ischaemic myocardium. PMID- 9415304 TI - Cystic fibrosis: gene therapy or preventive gene transfer? PMID- 9415305 TI - The 'adenobody' approach to viral targeting: specific and enhanced adenoviral gene delivery. AB - Recombinant adenoviruses have enormous potential as vectors for gene therapy. They have evolved an efficient method of infection and a wide host range but this leads to concerns about the specificity of gene delivery. In order to target an adenovirus type 5-based vector we have investigated an antibody approach. A virus neutralising scFv antibody fragment was isolated from a phage library and a C terminal fusion protein with epidermal growth factor (EGF) constructed. This fusion protein, or 'adenobody', bound both to the fibre protein of the adenovirus and to the EGF receptor (EGFR) on human cells, and was able to direct adenoviral binding to the new receptor. Using this system the efficiency of viral infection was markedly enhanced and was targeted to the EGFR. The adenobody-directed infection correlated with the level of EGF receptor expressed on the cells and could be blocked by competition with pure EGF. Peptide inhibition experiments suggest that infection is mediated directly through attachment to the EGFR and does not require penton-integrin interactions. This work shows that the 'adenobody' approach can enhance the efficiency as well as target adenoviral infection and has numerous potential applications for gene therapy. PMID- 9415306 TI - A GFP reporter system to assess gene transfer and expression in human hematopoietic progenitor cells. AB - Hematopoietic stem cells are widely recognized as attractive targets for gene therapy but current protocols to transduce these cells using recombinant retroviral vectors are inefficient. To evaluate optimization of retroviral transduction of hematopoietic stem cells and stability of gene expression in their progeny, the green fluorescent protein (GFP) was explored as a reporter. We first improved sensitivity of detection > 100-fold over that achieved previously by using a novel retroviral vector (termed MGIN) expressing a high level of an enhanced GFP gene. Primitive human hematopoietic cells bearing the CD34 surface antigen and lacking lineage differentiation markers (CD34+ Lin-) were transduced with the MGIN vector using a clinically applicable supernatant procedure. Under the conditions employed, > 75% of the target cells retained the CD34+ Lin- primitive phenotype after 4-5 days in culture, of those > or = 25% expressed a high level of GFP detectable by both flow cytometric analysis and fluorescence microscopy. When transduced cells were cultured in clonogenic progenitor assays, GFP fluorescence was readily detected in situ, indicating that GFP expression was stable and not detrimental to the differentiative potential of the transduced CD34+ Lin- cells. We conclude that GFP is effective as a vital marker to quantity retrovirus-mediated gene transfer into human hematopoietic and perhaps other types of stem/progenitor cells, and monitor gene expression during their subsequent cell lineage determinations. PMID- 9415307 TI - Induction of antigen-specific antitumor immunity with adenovirus-transduced dendritic cells. AB - Transduction of dendritic cells (DC) can result in presentation of tumor associated antigens and induction of immunity against undefined epitopes. The present studies demonstrate adenovirus (Ad)-mediated transduction of the beta galactosidase gene in mouse DC. Similar transductions have been obtained with the gene encoding the DF3/MUC1 tumor-associated antigen. We show that the Ad transduced DC are functional in primary allogeneic mixed lymphocyte reactions. Mice immunized with Ad-transduced DC develop cytotoxic T lymphocytes that are specific for the beta-galactosidase or DF3/MUC1 antigens. The results also demonstrate that Ad MUC1-transduced DC induce a specific response which inhibits the growth of DF3/MUC1-positive tumors. These findings support the usefulness of Ad-transduced DC for in vivo immunization against tumor-associated antigens. PMID- 9415308 TI - IFN-gamma gene transfer restores HLA-class I expression and MAGE-3 antigen presentation to CTL in HLA-deficient small cell lung cancer. AB - In this study, we have analyzed the possibility of inducing T cell responses against small cell lung cancer (SCLC), a still incurable tumor, by cytokine gene transfer approaches. By RT-PCR analysis most SCLC expressed the CTL-defined tumor antigens MAGE-3 (10/11), MAGE-1 (7/11) and less frequently BAGE (4/11) and GAGE1,2 (4/11). Although the surface expression of HLA class I molecules was low on most SCLC, thus preventing CTL recognition, treatment of the cells with IFN gamma enhanced HLA-class I levels in all cases. Two MAGE3+ SCLC cell lines displaying the A2 HLA-class I allele, involved in MAGE-3 antigen presentation to CTL, were stably transfected with the IFN-gamma gene (alone or co-transfected with IL-2). IFN-gamma-transfected cells displayed a clearcut increase in expression of HLA-class I and beta 2-microglobulin at both protein and mRNA level, and of TAP-1 and TAP-2 mRNA. Perhaps more importantly, IFN-gamma transfected cells were recognized by the MAGE-3-specific A2-restricted antimelanoma CTL clone 297/22, while unmodified cells or cells transfected with the IL-2 gene alone were not. These data indicate that IFN-gamma gene transfection into HLA-deficient SCLC cells is able to restore their ability to present endogenous tumor antigens to CTL and that IFN-gamma gene transfer approaches may be attempted to induce specific CTL responses in SCLC. PMID- 9415309 TI - Ornithine-delta-aminotransferase expression and ornithine metabolism in cultured epidermal keratinocytes: toward metabolic sink therapy for gyrate atrophy. AB - There is now strong evidence that the chorioretinal degeneration associated with ornithine-delta-aminotransferase (OAT) deficiency is a consequence of hyperornithinemia. Therefore development of a metabolic system for clearing ornithine from the circulation is being pursued as a potential treatment. The skin is considered an attractive location for such a metabolic system because autologous cells can be safely and easily utilized. This study was undertaken to determine the ornithine metabolizing capacity of epidermal keratinocytes expressing normal and superphysiologic amounts of OAT. The data show that overexpression of OAT in keratinocytes cultured from a gyrate atrophy patient restores ornithine metabolism and results in a rate of ornithine disappearance from the medium that is significantly higher than the rate of disappearance from the medium bathing normal keratinocytes. In addition, OAT activity determined in soluble protein prepared from sonicates suggests that the capacity to maintain plasma ornithine within the normal range is contained within an accomplishable graft of keratinocytes overexpressing OAT. However, the actual rate of ornithine disappearance from the media was significantly less than predicted from enzyme activity assays. Following ornithine metabolite production by intact cells suggests that ornithine metabolism is limited primarily by clearance of downstream metabolites, as opposed to substrate delivery. PMID- 9415310 TI - Use of transcriptional regulatory elements of the MUC1 and ERBB2 genes to drive tumour-selective expression of a prodrug activating enzyme. AB - In order to exploit differences in gene expression between normal and malignant cells for genetic prodrug-activation therapy, we have generated recombinant retroviruses containing the herpes simplex virus thymidine kinase coding region cloned downstream of sequences derived from the 5'-flanking regions of the MUC1 and ERBB2 genes. Transduction with retroviruses containing MUC1 promoters resulted in an increase in GCV sensitivity in MUC1 positive cells. A further increase in GCV sensitivity was achieved when MUC1-positive cells were transduced with retroviruses containing chimeric-MUC1/ERBB2 promoters. No significant sensitization to GCV was observed when MUC1-negative cells were transduced with these recombinant retroviruses. These results suggest that one may be able to develop a tumour-selective therapy by utilizing the transcriptional regulatory regions of the MUC1 and ERBB2 genes to drive the expression of suicide genes. PMID- 9415311 TI - Interferon-alpha gene therapy for cancer: retroviral transduction of fibroblasts and particle-mediated transfection of tumor cells are both effective strategies for gene delivery in murine tumor models. AB - Stable transfection of tumor cells with IFN-alpha genes has been shown to result in abrogation of tumor establishment and induction of antitumor immunity. However, strategies suitable for the clinical application of IFN-alpha gene therapy for cancer have not been reported. In this study, we investigated two gene delivery systems capable of mediating the local paracrine production of high levels of biologically active IFN-alpha in murine tumor models: retroviral transduction of fibroblasts and particle-mediated transfection of tumor cells. In spite of the antiproliferative effects of IFN-alpha, it was possible to obtain stable retroviral producer cell lines and transduce a variety of murine tumor cells including syngeneic fibroblasts to stably secrete 2000-5000 U (40-100 ng) murine IFN-alpha/10(6) cells/24 h. IFN-alpha transduction of tumor cells abrogated tumorigenicity in establishment models and induced antitumor immunity in several murine tumor model systems. Importantly, IFN-alpha gene delivery using retrovirally transduced syngeneic fibroblasts was capable of suppressing the establishment of the poorly immunogenic TS/A mouse mammary adenocarcinoma and induced antitumor immunity. Particle mediated transient transfection of tumor cells using the gene gun led to the production of up to 20,000 U IFN-alpha/10(6) cells during the first 24 h and proved to be equally effective in suppressing establishment of TS/A adenocarcinoma and inducing antitumor immunity. These results suggest that retroviral transduction of autologous fibroblasts can serve as an effective gene delivery method for IFN-alpha gene therapy of cancer. Particle-mediated transfection of freshly isolated tumor cells may represent a clinically attractive alternative approach for nonviral gene delivery. Both strategies circumvent the difficulties in routinely establishing primary tumor cell lines from the vast majority of human cancers. PMID- 9415312 TI - Double-copy bicistronic retroviral vector platform for gene therapy and tissue engineering: application to melanoma vaccine development. AB - The efficient genetic modification of solid tumors in situ to stimulate therapeutic immune responses against them is currently under active investigation, but is not yet possible using existing gene transfer technologies. Thus, ex vivo/in vivo vaccination strategies have been proposed in which the patient's tumor is surgically excised, single cell suspensions are prepared, the therapeutic genes are introduced and then the gene-modified cells, after being gamma-irradiated, are injected back into the patient. However, even with high efficiency gene delivery systems, this is a labor-intensive process. Moreover, it is often difficult to obtain sufficient numbers of gene-modified primary tumor cells during short-term culturing. On the other hand, extended in vitro passaging of primary tumor explants may alter their immunophenotypic properties. One approach to overcome these limitations would be to design universal vaccines consisting of standardized gene-transduced neoplastic cell lines or mixtures of gene-transduced cell lines to be combined with autologous tumor samples if available. Melanoma, which is notable for being one of the most immunogenic human malignancies, represents a cancer where shared tumor-associated antigens have been identified. We developed and analyzed several different retroviral vectors for their ability to stably express exogenous genes at high levels in a panel of melanoma cell lines. All vectors contained a reporter gene (nlslacZ) encoding beta-galactosidase with a nuclear localization signal and the neomycin phosphotransferase (neo) gene as selectable marker. One vector, DCCMV, which carried a bicistronic nlslacZ-neo transcriptional unit under the control of the human cytomegalovirus immediate-early promoter in the U3 region of its 3' LTR, was found to perform consistently better than the other vectors. The DCCMV vector, which is an extreme example of the double-copy class of retroviral vectors, was subsequently used to generate melanoma cell lines constitutively secreting human interleukin-6 or a soluble form of the human interleukin-6 receptor for potential use in a phase II clinical vaccine trial for the treatment of melanoma patients. The DCCMV vector design may also be useful in gene therapy applications where the intent is to implant polymer-encapsulated cell lines genetically engineered to stably express high levels of bioactive proteins. PMID- 9415313 TI - Pre-existing immunity to adenovirus does not prevent tumor regression following intratumoral administration of a vector expressing IL-12 but inhibits virus dissemination. AB - Adenovirus (Ad) vectors are being intensively studied as vehicles for cancer gene therapy. We have been exploring the benefits of direct intratumoral injection of Ads expressing cytokines for immunotherapy. Our previous work demonstrated that therapy using a vector expressing interleukin-12 (AdmIL-12.1) produced regressions in approximately 80% of treated tumors supporting further preclinical investigations. Recent reports have shown that immunity to Ad can be a major limiting factor in Ad-mediated gene transfer. As most animal studies with Ad vectors have involved nonimmune hosts, it remains difficult to predict how effective these treatments will be in humans, where the majority of individuals have had previous exposure to Ad. To address this question, we compared the effectiveness of the AdmIL-12.1 cancer therapy in naive and Ad-immune mice. We found that both groups responded equally well to treatment and that the response to AdmIL-12.1 in both groups resulted in the generation of CTL reactive against tumor antigen indicating that antitumor immunity was achieved. Peak transgene expression in the tumor was only reduced by 2.4 fold in Ad-immune animals compared with nonimmune mice. It was also observed that in naive animals, the virus disseminated from the site of the tumor following injection and by 72 h substantial transgene expression was detected in peripheral organs, most notably the liver. Transgene expression in the liver of Ad-immune animals was reduced by greater than 1000-fold relative to that in naive mice. These results strongly support the clinical utility of Ad-based cancer gene therapy and suggest that Ad immunity may be advantageous in that it is not a complete block to gene transfer in the tumor and it greatly reduces virus dissemination. PMID- 9415314 TI - Retrovirus-mediated gene transfer corrects DNA repair defect of xeroderma pigmentosum cells of complementation groups A, B and C. AB - With the aim to devise a long-term gene therapy protocol for skin cancers in individuals affected by the inherited autosomal recessive xeroderma pigmentosum we transferred the human DNA repair XPA, XPB/ERCC3 and XPC cDNAs, by using the recombinant retroviral vector LXSN, into primary and immortalized fibroblasts obtained from two XP-A, one XP-B (associated with Cockayne's syndrome) and two XP C patients. After transduction, the complete correction of DNA repair deficiency and functional expression of the transgenes were monitored by UV survival, unscheduled DNA synthesis and recovery of RNA synthesis, and Western blots. The results show that the recombinant retroviruses are highly efficient vectors to transfer and stably express the human DNA repair genes in XP cells and correct the defect of DNA repair of group A, B and C. With our previous results with XPD/ERCC2, the present work extends further promising issues for the gene therapy strategy for most patients suffering from this cancer-prone syndrome. PMID- 9415315 TI - Aberrant, noninfectious HIV-1 particles are released by chronically infected human T cells transduced with a retroviral vector expressing an interfering HIV-1 variant. AB - The expression of the nonproducer F12-HIV-1 genome has been previously shown to protect the host cell from HIV superinfection. In order to estimate the efficacy of the F12-HIV genome as an anti-HIV reagent also in cells already infected, an HIV-1 chronically infected Hut-78 cell clone (D10) was superinfected with an amphotropic mouse/human pseudotype retrovirus whose genome expresses both the F12 HIV genome and the selection marker gene (i.e. the c-DNA of a truncated form of the nerve growth factor receptor, NGFr) under the control of F12-HIV 5'LTR. D10 cells homogenously expressing the F12-HIV genome (T-D10) released unaltered amounts of retrovirions whose infectivity was, however, dramatically impaired (from 9 x 10(3) in D10 to < 10(0.5). TCID50/ml in T-D10 supernatants). Electron microscopy showed that the morphology of retrovirions released by T-D10 cells was heavily altered, both in size and shape. Furthermore, no retrotranscription products were detectable in CD4 cells challenged with T-D10 retrovirions. For the first time, the block in the infectivity of HIV released from already infected cells through the expression of an anti-HIV retroviral vector was demonstrated. These data could have important implications both from a perspective of F12-HIV based anti-HIV gene therapy and, in general, on the role that nonproducer and/or defective HIV could play 'in vivo' in HIV infection and AIDS pathogenesis. PMID- 9415316 TI - Efficient adenovirus-mediated gene transduction of normal and leukemic hematopoietic cells. AB - We evaluated the efficiency of adenovirus-mediated gene transfer into normal and malignant human hematopoietic cells. An E-1 and E-3 deleted, replication defective recombinant Ad.RSV beta gal vector was used and the transduction efficiency was studied at a multiplicity of infection of 13 p.f.u. per cell. Approximately 40-50% of normal monocytes were transduced, whereas purified normal resting T cells and B cells were resistant to infection. We showed that 50-80% of primary chronic myeloid leukemia cells (CML, n = 12) were efficiently transduced in contrast to CML, successful transduction of resting primary chronic B lymphocytic leukemia cells required appropriate preactivation of targeted cells. A novel protocol for the efficient transduction of adenovirus into B-CLL cells was presented. We showed that anti-CD40 mAb or CD40 ligand acts in synergy with rhIL-4 to enable the transduction of approximately 50-75% of B-CLL cells (B-CLL, n = 6). Expression of beta-galactosidase in transduced CML cells and B-CLL cells was detected for at least 15 days after transduction. The present studies underline the utility of adenovirus vectors for the construction of cytokine gene modified tumor vaccines for the treatment of hematopoietic malignancies such as CML and B-CLL. PMID- 9415317 TI - ExGen 500 is an efficient vector for gene delivery to lung epithelial cells in vitro and in vivo. AB - Nonviral vectors might represent a safe alternative to adenovirus for gene therapy of lung disorders, in particular cystic fibrosis (CF). Cationic lipids have been shown to correct the CF defect both in vitro and in vivo, but more efficient vectors are needed to improve the low gene transfer efficiency. Here, we show that the cationic polymer ExGen 500, a linear polyethylenimine derivative, is more efficient than cationic lipids in transferring reporter genes to lung epithelial cells in vitro. In vivo ExGen 500 was able to mediate gene transfer into both newborn and adult rabbit lungs with comparable efficiencies. The best levels of transfection were obtained using neutral complexes. Under such conditions, luciferase activities corresponding to about 10(3) RLU/10 s/mg of protein were reproducibly obtained 2 days after transfection throughout the four lung lobes of newborn and adult rabbits. A nlslacZ reporter gene showed transfected cells around the lumen of large and small bronchi. No signs of acute toxicity (inflammation, cellular infiltration etc) were detected by direct histopathological analysis. Within 1 week after instillation, transgene expression decreased by two orders of magnitude. PMID- 9415318 TI - Varicella-zoster virus thymidine kinase gene and antiherpetic pyrimidine nucleoside analogues in a combined gene/chemotherapy treatment for cancer. AB - Ten pyrimidine nucleoside analogues, including (B)-5-(2-bromovinyl)-2' deoxyuridine (BVDU) and closely related analogues, were evaluated for their cytostatic activity against human osteosarcoma cells transfected with the varicella-zoster virus (VZV) thymidine kinase (tk) (ATP:thymidine 5' phosphotransferase, EC 2.7.2.21) gene. (E)-5-(2-bromovinyl)-1-beta-D arabinofuranosyluracil (BVaraU), (E)-5-(2-iodovinyl)-2'-deoxy-2'-fluoro-1-beta-D arabinofuranosyluracil (IVFAU) and (E)-5-(2-bromovinyl)-2'-deoxy-4'-thiouridine (S-BVDU) were among the most potent inhibitors of VZVtk gene-transfected cell proliferation. They displayed an inhibitory activity at drug concentrations that were up to four orders of magnitude lower than those required to inhibit the corresponding nontransfected tumor cells. Inhibition of cellular DNA polymerase and/or incorporation of the drugs into cellular DNA may be a likely target for the cytostatic activity of the BVDU derivatives against the VZVtk gene transfected tumor cells. These compounds were approximately 40- to 80-fold more potent cytostatic agents in VZVtk gene-transfected cells than the anti-VZV compound 6-methoxy-9-beta-D-arabinofuranosylpurine (araM), and at least five- to 50-fold more cytostatic than ganciclovir in HSV-1tk gene-transfected murine mammary carcinoma FM3A cells. In addition, the intrinsic resistance of BVaraU, IVFAU and S-BVDU to glycosidic bond cleavage by mammalian dThd phosphorylases makes them promising candidate compounds for the treatment of VZVtk gene transfected tumors in vivo. PMID- 9415319 TI - The internal ribosomal entry site of the encephalomyocarditis virus enables reliable coexpression of two transgenes in human primary T lymphocytes. AB - It is essential for the improvement of adoptive cell therapies to generate efficiently large populations of human primary T cells that reliably express a suicide gene conferring drug sensitivity, such as herpes simplex virus thymidine kinase (HSVtk). We show here that an optimized dicistronic vector containing the encephalomyocarditis virus (EMCV) internal ribosomal entry site (IRES) is functional in human primary. T lymphocytes that bear on average one integrated vector copy per cell. We demonstrate reliable coexpression of the marker NTP, an inactive mutant of the human low-affinity nerve growth factor receptor and HSVtk. In the dicistronic vector NIT, NTP is expressed as a cap-dependent marker and HSVtk as a nonselectable IRES-dependent gene. Cell-surface expression of NTP is sufficient to allow for the efficient and rapid enrichment of the transduced cells to high purity. Of these purified lymphocytes, 97 +/- 4% and 92 +/- 6% are selectively eliminated when cultured in the presence of 1.0 or 0.1 microM ganciclovic respectively, establishing that the EMCV IRES ensures efficient and sufficient expression of two genes in human primary T cells. PMID- 9415320 TI - Rapid method for construction of recombinant HSV gene transfer vectors. AB - Herpes simplex virus type 1 (HSV-1) is a neurotrophic human pathogen that naturally persists in neurons in a latent state and carries a large number of viral functions which can be replaced by foreign genes to create a vector for gene therapy applications. In this report we describe a two-step method for insertion/deletion mutagenesis of HSV genes and the efficient insertion of transgenes into these locations in the viral genome. The first step is the insertion of a reporter gene (lacZ) cassette flanked by Pacl restriction enzyme sites not otherwise found in the viral genome, using standard marker transfer procedures to interrupt a portion of the target HSV gene. The second step is substitution of the reporter gene with other foreign cDNAs by digestion of the vector DNA with Pacl to remove the lacZ gene and subsequent repair of the vector genome by homologous recombination with a transgene expression plasmid. Potential recombinants identified by a 'clear plaque' phenotype after X-gal staining arose at high frequency (80-100%). Of these recombinants containing the transgene in place of the lacZ gene ranged from 19-65%. Insertion of the transgene expression construct into the viral genome eliminates the Pacl sites, allowing this method to be used repeatedly for the sequential deletion of multiple HSV genes while inserting multiple transgenes. This procedure was repeated in succession to produce a vector carrying two independent expression cassettes at distinct viral loci. PMID- 9415321 TI - Encoding activity and face recognition. AB - A series of studies conducted over the past 20 years have explored the effects of various tasks on recognition memory for faces. Memory for faces appears better when the study task involves judgements about an abstract trait rather than a physical feature. The various situations in which these results were obtained raise important methodological questions regarding the learning conditions, whether incidental or intentional, and the duration of exposure to the stimulus during the study phase. We consider here two alternative explanations for the reported results. One concerns depth of processing and the other the opposition between component and holistic processing. Possible strategies for improving face recognition performance are considered. PMID- 9415322 TI - Multiple organisations of events in memory. AB - Theories of memory organisation propose that activity knowledge organises autobiographical memory globally. According to these views, memories that share a participant, location, or time are only organised together if they also share an activity. If they do not, they are nested within their respective activity organisations locally rather than being organised together globally. Two experiments that assessed people's clustering of laboratory events consistently obtained findings that contradict this view. Both experiments found that people organise event memories globally in non-activity clusters, cross-classify events into multiple organisations, and pivot between activity and non-activity clusters. Consistent with studies of naturalistic events, these studies of laboratory events indicate that people cross-classify event memories simultaneously into multiple global organisations. PMID- 9415323 TI - Cultural factors in the recall of a witnessed event. AB - Can cultural factors influence testimony? To explore this hypothesis, groups of Spanish and English students (n = 48 per group) observed successively two films showing events specific to Spain (a romeria) and England (a village fete) before giving free recall and a verbal recognition test that contained lures consistent and inconsistent with cultural expectations. In addition, half the subjects were instructed to adopt the role of an observer while the remainder played the role of a participant. Significant interactions between cultural background and type of event reported were found for both recall and recognition. Contrary to expectation (i) recall accuracy was greater for the event that was not from the subject's own country (ii) recall errors were greater for the event from the subject's own country. In line with expectation, recognition accuracy was high for the event from the subject's own country. Results are discussed in terms of the operation of schema-based theories of recall and Johnson's reality-monitoring approach. The practical implications of the current findings for the interviewing of witnesses are briefly discussed. PMID- 9415324 TI - Memories for the Marchioness. AB - We examined the accounts of 27 survivors of the Marchioness ferry sinking, using cross-validation of accounts to search for instances of motivated forgetting. In order to identify objective items that could be validated, we focused the analysis on subjects' statements of whom they were with at various stages of the disaster. We compared these findings with an informal recall of a non-traumatic event after an interval of a few days. The main finding was that recall was reasonably good for both traumatic and non-traumatic events. Specifically, in the Marchioness sample, among those 86 statements that could have been confirmed in the accounts of other informants, 74 were in fact confirmed. Of the remaining 12 unconfirmed statements, only one involved a contradiction. We conclude that for a disaster of this kind, and with this particular sample of individuals, motivated forgetting was extremely rare. PMID- 9415325 TI - Eyewitness performance in cognitive and structured interviews. AB - This paper addresses two methodological and theoretical questions relating to the Cognitive Interview (CI), which previous research has found to increase witness recall in interviews. (1) What are the effects of the CI mnemonic techniques when communication techniques are held constant? (2) How do trained interviewers compare with untrained interviewers? In this study, witnesses (college students) viewed a short film clip of a shooting and were questioned by interviewers (research assistants) trained in conducting the CI or a Structured Interview (SI) -similar to the CI except for the "cognitive" components--or by untrained interviewers (UI). The CI and SI groups recalled significantly more correct information compared to the UI group. However they also reported more errors and confabulated details. Theoretical and practical implications of the results are discussed in terms of precisely identifying the CI facilitatory effects and consequent good practice in the forensic setting. PMID- 9415326 TI - Oral leukoplakia: a clinicopathological review. AB - Leukoplakia is the most common premalignant or potentially malignant lesion of the oral mucosa. It seems preferable to use the term leukoplakia as a clinical term only. When a biopsy is taken, the term leukoplakia should be replaced by the diagnosis obtained histologically. The annual percentage of malignant transformation varies in different parts of the world, probably as a result of differences in tobacco and dietary habits. Although epithelial dysplasia is an important predictive factor of malignant transformation, it should be realized that not all dysplastic lesions will become malignant. On the other hand non dysplastic lesions may become malignant as well. In some parts of the world the tongue and the floor of the mouth can be considered to be high-risk sites with regard to malignant transformation of leukoplakia, while this does not have to be the case in other parts of the world. The cessation of tobacco habits, being the most common known aetiological factor of oral leukoplakia, has been shown to be an effective measure with regard to the incidence of leukoplakia and, thereby, the incidence of oral cancer as well. Screening for oral precancer may be indicated in individuals at risk. PMID- 9415327 TI - Epidemiology and prevention of oral cancer. AB - Descriptive epidemiology of oral and pharyngeal cancer over the last four decades is reviewed, with specific focus on Europe. Substantial rises in mortality rates have been observed for younger males, mostly in eastern Europe. The independent role of alcohol and tobacco and their interaction on oral carcinogenesis is discussed, since these factors account for about three quarters of oral cancers in Europe. The influence of dietary factors, and in particular of a diet poor in fresh fruit and vegetables on oral carcinogenesis, is also discussed, since diet may account for 10-15% of oral cancer cases in Europe. Finally, among other carcinogens, the possibility of human papillomavirus involvement in the aetiology of cancer of the oral cavity and pharynx is overviewed. Implications for prevention are discussed. PMID- 9415328 TI - Screening for cancer of the head and neck: if not now, when? AB - There is as yet no evidence to support population screening for oral cancer, although the mouth is easy to examine, and the disease is common in certain parts of the world and/or subsegments of the population. Oral cancer screening programs have been carried out on several hundred thousands of individuals in developing countries (mostly India and Cuba) and several thousands in developed countries (mostly the U.S.A., U.K. and Italy). Especially in developed countries, lesions of the pharynx and larynx were also searched for. Substantial portions of individuals with suspicious lesions (around 10%), mostly leukoplakia, could be identified, but major difficulties were found in targeting highest-risk individuals and referring them to a specialised centre, when necessary. When oral inspection was repeated, relatively high incidence of oral cancer, after removal of prevalent cases, suggested a rather short sojourn time for preclinical cancer (in the order of one year). Oral cancer screening programmes would be greatly facilitated by screening tests able to anticipate the detection of a preclinical phase, compared to visual inspection, thus allowing screening intervals to be prolonged. Finally, even if dysplastic lesions of the oral cavity were better recognised and understood (e.g. as for intra-epithelial lesions of the cervix uteri), surgical control of the disease would be harder than for the uterus, breast, or colon-rectum. PMID- 9415329 TI - Oral leukoplakias: to treat or not? PMID- 9415330 TI - Local antimicrobial therapy of oral mucositis in paediatric patients undergoing bone marrow transplantation. AB - The present investigation has examined the clinical benefits of tobramycin, polymyxin E and amphotericin therapy in the management of oral mucositis in children undergoing chemotherapy prior to bone marrow transplantation. Tobramycin, polymyxin E, and amphotericin reduced the degree of oral mucositis more than conventional therapy of diphenhydramine, Maalox, and local analgesic. While there was a statistically significant fall in the severity of the mucositis with tobramycin, polymyxin E and amphotericin, this was unlikely to be of practical benefit. PMID- 9415331 TI - Risk factors for salivary dysfunction in children 1 year after bone marrow transplantation. AB - Bone marrow transplantation (BMT) in childhood has improved the survival for children with leukaemia, severe aplastic anaemia and some metabolic disorders. However, transplant procedures are associated with a high incidence of deleterious long-term complications. This study included 49 children, conditioned according to the Seattle protocols. Follow-up oral examinations were performed 1 year after BMT. Forty risk factors that may have influenced a low stimulated salivary secretion rate (SSSR) were studied. An SSSR < or = 0.5 ml/min, 1 year after BMT was regarded as the event. One year after BMT, the mean SSSR was 0.5 +/ 0.4 ml/min in children conditioned with total body irradiation (TBI) compared to 1.0 +/- 0.5 ml/min in non-irradiated children (P = 0.0013). Significant predictors for low SSSR in multivariate analysis were: conditioning with TBI (P = 0.0023), recipient female sex (P = 0.0121) and recipient seropositivity for between three and four herpes viruses (P = 0.0157). PMID- 9415332 TI - Field cancerisation of the oral cavity: comparison of the spectrum of molecular alterations in cases presenting with both dysplastic and malignant lesions. AB - Dysplastic lesions and invasive oral squamous cell carcinoma (SCC) from patients with field change were screened by restriction fragment length polymorphism (RFLP) and microsatellite assay. All tumours contained more genetic changes than the matched dysplasia which are likely to represent progression. Four of the 15 dysplastic lesions harboured the same abnormalities detected in the tumour and some paired lesions showed identical novel microsatellite alleles. The finding of identical 'genetic fingerprints' in dysplastic lesions and invasive carcinoma from the same patient provides strong evidence that these dysplasias are precursor lesions and that multiple lesions have probably arisen due to transfer of the progeny of an altered cell. Eight of the 15 dysplastic lesions showed alterations which were not present in the matched cancer, showing that evolution of subclones, or fusion of multiple clones also occurs. A further case showed loss of different alleles in the paired samples. These findings highlight the complexity of the genetic abnormalities present in the mucosa of patients with field change and suggests that the origin of these altered foci may be diverse. PMID- 9415333 TI - Mouthwash use and dentures in relation to oral epithelial dysplasia. AB - This case-control study investigated the potential association between oral epithelial dysplasia (OED) and both mouthwash and denture use. Incident OED cases aged 20-79 years were identified through two oral pathology laboratories. Controls were pair-matched (1:1) to cases on age (+/- 5 years), gender, appointment date and surgeon. A telephone interview was used to obtain exposure information. Odds ratios (ORs) and 95% confidence intervals (95% CIs) were generated using conditional logistic regression. Based upon 127 case-control pairs and after adjusting for smoking, drinking, education and either denture or mouthwash use, the OR for OED and regular mouthwash use (1+ uses/week for 6+ months) was 0.8 (95% CI, 0.4-1.5) while the OR for OED and wearing a denture was 0.7 (95% CI, 0.4-1.3). There were no clear trends of increased OED risk with increased mouthwash use or years of denture wearing. Our findings suggest that neither mouthwash nor denture use are associated positively with OED risk. PMID- 9415334 TI - Cytogenetic characterisation of Warthin's tumour. AB - Warthin's tumour is a peculiar subtype of monomorphic adenomas of the salivary glands, frequently cystic, and that characteristically associates an epithelial glandular cell component to a dense lymphoid infiltrate. Short-term cultures from 12 Warthin's tumours of salivary glands, including 5 previously reported cases were successfully karyotyped and clonal numerical and/or structural changes were detected in 7 of them (58%). 3 cases showed numerical abnormalities with loss of chromosomes Y (2 cases) and X (1 case). The remaining 4 abnormal cases presented the following structural changes: complex translocation t(11;19;16)(q21;p12;p13.3); reciprocal translocations t(6;8)(p23;q22) and t(6;15)(p21;q15) (2 cases); and 1p22, 3p26, 11p13 changes. In 1 case, clonal numerical deviations (+ 7 and -Y) were concurrent with the structural rearrangement t(6;8). Two of these aberrations are suggested to be Warthin's tumour-associated: 11q;19p translocation has already been described in 3 cases, and structural rearrangements of 6p23 have also been reported in another case. Our study extends the cytogenetic information about Warthin's tumour and identifies two recurrent abnormalities --6p rearrangements and t(11;19)--specific for this salivary neoplasm. PMID- 9415335 TI - Dental parameters in the long-term survivors of childhood cancer compared with siblings. AB - There have been a number of reports on the dental health of long-term survivors (LTS) of childhood malignancy as compared with normal controls. However, it is usually difficult to identify a meaningful control population as most of these patients are from widely differing geographical areas and socio-economic status. The aim of this investigation was, therefore, to study the dental health of LTS compared with siblings. 46 LTS who had siblings of a similar age were identified for the study. Both groups were examined for dental caries, gingivitis and enamel defects. There was no statistically significant difference in the mean DMFS of the test and control groups. However, the LTS had a significantly (P = 0.006) higher number of decayed surfaces (1.50 +/- 0.30) as compared with their siblings (0.50 +/- 0.20). The LTS also had a significantly higher prevalence of severe gingivitis (1.11 +/- 0.33) compared with controls (0.02 +/- 0.02). There was a significantly higher prevalence of all types of enamel defects in the LTS and fewer teeth with no enamel defect as compared with their siblings, with the mean values being 15.7 +/- 0.9 and 25.3 +/- 0.3, respectively. It was concluded that there was a higher prevalence of untreated dental disease and developmental defects in long-term survivors. PMID- 9415336 TI - Ondansetron suppository: an effective treatment for the prevention of emetic disorders induced by cisplatin-based chemotherapy. French Ondansetron Study Group. AB - This multicentre, randomised, double-blind, double-dummy, parallel group study was investigated in order to compare on 3 days the efficacy and the safety of the 16 mg once a day (od) ondansetron suppository (suppository group) with the recommended ondansetron treatment, i.e. 8 mg intravenous (i.v.) ondansetron on day 1 followed by 8 mg tablet (p.o.) twice daily (i.v. + p.o. group) on days 2 and 3 in patients receiving cisplatin (> or = 50 mg/m2) containing chemotherapy. In the 420 patients included in the intent-to-treat population, 209 received the 16 mg suppository and 211 the i.v. + p.o. treatment. The number of emetic episodes and the nausea score were recorded each day. Concerning the primary criterion, both treatments provided good anti-emetic control with 87% of all patients having a complete or major response (0-2 emetic episodes) on day 1 in the suppository group and 92% in the i.v. + p.o. group (P = 0.058). The 90% confidence interval for the difference between the two treatments for complete or major control was included in the interval (-15%, 15%) and showed that the treatment groups could be regarded as equivalent. Small differences in favour of the i.v. + p.o. group were observed concerning the secondary parameters. Both treatments were well tolerated. The results of this study show that both treatments are equivalent in the prevention of cisplatin-containing chemotherapy induced emesis for the primary efficacy criteria and that the ondansetron suppository is efficient and well tolerated and is a suitable alternative to the anti-emetic treatment combining the intravenous and oral routes. PMID- 9415337 TI - Assessment of epidermal growth factor in oral secretions of patients receiving radiation therapy for cancer. AB - Biological response modifiers have been studied in animal models of oral mucositis. We assessed the presence of epidermal growth factor (EGF) in patients during radiation therapy for head and neck cancer. The findings of this preliminary study showed that it is possible to measure the presence of EGF in oral secretions during radiation therapy. EGF was shown to decrease during the course of radiation therapy, and a trend was seen with decreasing EGF and increasing oral ulceration (P = 0.10) and increasing total mucositis score (P = 0.09). PMID- 9415338 TI - Nutrient intake and weight development in children during chemotherapy for malignant disease. AB - The aim of the study was to assess the actual daily oral intake of energy, protein, fat and carbohydrate in relation to current recommendations in children with malignant disease during chemotherapy and to follow their weight development. Dietary information was collected for 21 consecutive days via 7-day recording in 14 children, aged 5-16 years. The number of days with loss of appetite, vomiting, and the number of days on anti-emetic drugs were also recorded. The average daily energy intake decreased from 91% of the recommendation of the Swedish Nutrition Recommendations (SNR), before chemotherapy to 69% after start of chemotherapy. During days spent at home, the energy intake increased to 77% of SNR. Twenty-two per cent of the total energy intake during the hospital days came from sucrose. On average, the children experienced loss of appetite on 50% of the days, vomiting on 12%, and received anti-emetic drugs on 38%. On admission, the average SD score for body weight for the whole group was -0.09. The mean weight reduction after 1 week was 0.19 SD (P = 0.05) compared to the admission weight. The weight reduction 6 weeks (n = 10) and 3 months (n = 13) after the start of chemotherapy was 0.10 SD and 0.37 SD (P = 0.04), respectively. PMID- 9415339 TI - Immunohistochemical study of tumour angiogenesis in oral squamous cell carcinoma. AB - To assess the clinical significance of angiogenesis in oral squamous cell carcinoma (SCC), we examined vessel density immunohistochemically in 44 primary oral SCCs using the JC-70A antibody which reacts specifically with vascular endothelial cells. In addition, the expression of vascular endothelial growth factor (VEGF) and its receptors, KDR, Flt-1 and Flt-4 in oral SCCs was examined in relation to the vessel density and lymph node metastasis. There was no association of vessel density with tumour site, T-category (tumour size), degree of differentiation or cervical lymph node metastasis, except that the vessel density of carcinomas with a well-defined tumour-stromal boundary was higher than that of diffusely invasive carcinomas. The intensity of VEGF expression correlated with lymph node metastasis (P < 0.01), but not with vessel density. The expression of KDR and Flt-1 did not correlate with vessel density and lymph node metastasis. However, the vessel density in Flt-4-positive carcinomas was higher than that in Flt-4-negative carcinomas (P < 0.05), and expression of Flt-4 most significantly correlated with lymph node metastasis (P < 0.001). These results suggest that the expression of VEGF or Flt-4 rather than vessel density may be a predictor of lymph node metastasis in oral SCC. PMID- 9415340 TI - Traumatic eosinophilic granuloma of the oral mucosa: a CD30+(Ki-1) lymphoproliferative disorder? AB - Traumatic eosinophilic granuloma of the oral mucosa, also known as eosinophilic ulcer, is considered to be a reactive lesion of unknown aetiology. It usually presents as a tongue ulcer and injury has been considered to play a role in its cause. We present a 72-year-old man who had suffered multiple episodes of recurrent eosinophilic ulcers of the oral mucosa which underwent self-healing. Biopsy specimens (including fresh tissue) were studied with a combination of histology, electron microscopy and immunohistochemistry. A dense cell infiltrate composed of eosinophilis, lymphocytes and large mononuclear cells was constantly shown. Immunostains showed that the infiltrate was mainly composed of CD3+,CD4+,CD8-T-cells and CD1a + dendritic cells. Approximately 70% of the T cells expressed CD30 (Ki-1) antigen. On the basis of the clinical behaviour, histology and antigenic features, it seems reasonable to suggest that traumatic eosinophilic granuloma of the oral mucosa may represent the oral countpart of primary cutaneous CD30 (Ki-1)-positive lymphoproliferative disorders. This group of cutaneous lymphomas are indeed characterised by non-aggressive clinical behaviour (sequential evolution in ulceration, necrosis and self-regression) and expression of CD30 antigen by the infiltrating large T-cells. PMID- 9415341 TI - Chronic atrial fibrillation: optimizing anticoagulation. PMID- 9415342 TI - The features and management of poisoning with drugs used to treat Parkinson's disease. PMID- 9415343 TI - Pre-AIDS deaths in HIV infection related to intravenous drug use. AB - We prospectively collected data on deaths in the Edinburgh City Hospital HIV cohort of patients (60-70% acquired via injection drug use) from October 1986 to September 1994. Sixty-four patients (25% of all HIV deaths or 2.5/100 person years) had died without an AIDS diagnosis, and 42 (66%) of these had autopsy data available. Some pre-AIDS deaths (20% or 0.5/100 person-years) were the expected consequence of underlying medical conditions diagnosed during life: the remainder (80% or 1.98/100 person-years) were sudden or unexpected. Examining the underlying conditions, drug overdoses accounted for 45% or 1.1/100 person-years; bacterial sepsis, 25% or 0.6/100 person-years; liver disease, 26% or 0.6/100 person-years; and an undiagnosed AIDS condition, 9% or 0.2/100 person-years. Drug overdoses were the commonest cause of pre-AIDS death in this cohort of patients predominantly infected via IDU, but many of the sudden deaths had significant underlying pathology, which may have increased their susceptibility to an overdose of drugs. In future, death before an AIDS diagnosis should be classified into Medical or Expected Non-AIDS (MNA or ENA) and Sudden Non-AIDS (SNA). PMID- 9415344 TI - Fenofibrate plus simvastatin therapy versus simvastatin plus cholestyramine therapy for familial hypercholesterolaemia. AB - Combination therapy is routinely used to achieve improved cholesterol reduction in familial hypercholesterolaemia. We compared the standard simvastatin plus bile acid sequestrant (cholestyramine) therapy with simvastatin plus fenofibrate in 29 patients with severe familial hypercholesterolaemia. The fibrate regimen resulted in an 35.1 +/- 10.7% reduction in total cholesterol, a 40.6 +/- 20.5% in LDL cholesterol, 17.2 +/- 56.5% reduction in triglycerides and a 20.3 +/- 52.0% increase in HDL cholesterol. The cholestyramine regimen produced reductions of 29.3 +/- 13.2% in cholesterol, 37.1 +/- 21.9% in LDL cholesterol, and 12.5 +/- 48.9% in triglycerides, and a 5.0 +/- 25.4% rise in HDL cholesterol. The fibrate regimen was significantly more effective in reducing total cholesterol (p < 0.001) and LDL-cholesterol (p = 0.004), and also reduced triglycerides significantly (p = 0.05), compared to the cholestyramine regimen. There were significant improvements in the LDL:HDL cholesterol ratio (3.62 +/- 1.54 vs. 4.00 +/- 1.36; p = 0.05) and in the apolipoprotein B:A1 ratio (1.13 +/- 0.036 vs. 1.20 +/- 0.34; p = 0.05). Gastrointestinal side-effects occurred in 10 patients on cholestyramine therapy, and four patients on fibrate therapy had myalgia. There were no cases of rhabdomyolysis with either regime. No significant differences in liver biochemistry or creatine kinase were seen with either regimen. PMID- 9415346 TI - Donor sepsis is not a contraindication to cadaveric organ donation. AB - Systemic donor infection is regarded as being an absolute contraindication to cadaveric organ donation for transplantation. This is largely due to fear of transmitting pathogenic organisms to the immunosuppressed recipient. However, due to the current shortage of organs available for transplantation, clinicians are faced with the option of using organs from 'non-ideal' donors, such as those patients with documented evidence of infection. We report the successful outcome of six orthotopic liver transplants, 11 renal transplants, one combined heart lung transplant and one simultaneous kidney and pancreas transplant with organs from eight donors in whom bacterial meningitis (n = 7) and acute bacterial epiglottitis (n = 1) were the antecedent causes of death. PMID- 9415345 TI - Hyperhomocysteinaemia in black patients with cerebral thrombosis. AB - Hyperhomocysteinemia is regarded as a risk factor for stroke but its pathogenetic role has not yet been established in Black patients. We studied 24 Black patients admitted with cerebral thrombosis, and compared them with age- and sex-matched apparently healthy controls from the same community. Total homocysteine (tHcy) (free homocysteine, protein-bound homocysteine, the disulfide homocystine and the mixed disulfide homocysteine-cysteine) concentration was 10.91 (4.95-23.05) mumol/l in the stroke patients and 8.73 (3.95-15.10) mumol/l in controls (p = 0.031). This difference could not be explained by differences in vitamin B12, vitamin B6 or folate status. A subgroup of nine stroke patients with hypercreatininaemia (> 90 mumol/l, 75% of control concentrations) had significantly higher plasma tHcy concentrations [median (range) 9.10 (5.40-15.10) mumol/l] compared with controls [8.65 (3.96-13.89) mumol/l] (p = 0.002). Plasma tHcy concentrations of stroke patients with normal serum creatinine concentrations were not significantly different to those of controls. Hyperhomocysteinemia in Black patients with stroke may be partially caused by renal insufficiency. Therefore, while hyperhomocysteinemia may increase the risk of stroke, it is unlikely to be a primary initiating factor. PMID- 9415347 TI - Attacks of pericarditis as a manifestation of familial Mediterranean fever (FMF). AB - Familial Mediterranean fever (FMF) is characterized by recurrent attacks of febrile serositis. While arthritis, pleuritis and peritonitis are common in FMF, no association of pericarditis with FMF has been described in detail. We retrospectively studied about 4000 FMF patients, using a computer chart review. Pericarditis was diagnosed when patients sustained attacks of pleuritic retrosternal chest pain and had typical findings in the electrocardiogram, echocardiogram or chest radiogram. The incidence and features of pericarditis in FMF were compared to published data. Over a period of 20 years, one or more episodes of pericarditis were recorded in 27 patients, a significantly higher incidence than in the general population (68 vs. 6 per 10(5) per year, p < 0.001). Each patient experienced 1-3 pericarditis attacks, lasting a mean of 4.2 days, accompanied by high temperature and symptoms of FMF attack at another site. The pericarditis attack resolved spontaneously and left no sequelae. FMF patients with pericarditis were comparable to other FMF patients in most demographic and clinical parameters. Pericarditis may be considered another rare manifestation of FMF. PMID- 9415348 TI - Estimating the cost effectiveness of screening tests. AB - A number of criteria have been postulated that a screening test should fulfill in order for it to be a good or worthwhile screening tool. However, it is a rare test which fulfills all the criteria that have been thought to be important. Therefore, there always exists some form of trade-off between, for example, the detection rate and the false-positive rate and cost. It is the magnitude and form of these trade-offs, which differ for each screening test, that are important, not whether a test fulfills some arbitrary set of criteria. In this paper, we describe some of the trade-offs which occur when appraising a screening test and we argue that listing arbitrary criteria is an unhelpful activity. PMID- 9415349 TI - Propofol for sedation in the intensive care unit: essentials for the clinician. AB - Propofol is a short-acting intravenous anesthetic commonly utilised in the intensive care unit (ICU) for sedation of mechanically ventilated patients. The rapid onset and termination of action make it an attractive drug for use in the ICU. The safety profile of propofol is well established. However, there are potential adverse reactions associated with the drug. This review discusses the pharmacology, administration and adverse effects associated with propofol with which clinicians who administer propofol should be familiar. PMID- 9415350 TI - Hepatocyte growth factor and neutrophil elastase in idiopathic pulmonary fibrosis. AB - It has been hypothesized that hepatocyte growth factor (HGF) may play an important role in regulating the growth of lung epithelium and in the regeneration of the lung as a paracrine or endocrine factor in idiopathic pulmonary fibrosis (IPF). Based on this background, serum HGF was measured in 31 IPF patients (21 male/10 female, median age 60 years). Fifteen age-matched normal non-smokers served as the control. Hepatocyte growth factor was measured by enzyme-linked immunosorbent assay with monoclonal and polyclonal antibodies against human HGF (Otsuka Assay Laboratories, Tokushima, Japan). Elastase: alpha 1-proteinase complex was also measured by enzyme-linked immunosorbent assay. No patients had significant liver or renal dysfunction. As a result, mean (standard error) serum HGF concentration of the patients with IPF was 0.384 (0.022) ng ml 1, which was significantly high compared to normal non-smokers [0.213 (0.012) ng ml-1, P < 0.001, 95% confidence interval was between 0.104 and 0.238]. Serum HGF values correlated strongly with the plasma elastase: alpha 1 proteinase inhibitor complex (R = 0.679, P < 0.001). Immunohistochemical staining of lung tissue with anti-human neutrophil elastase showed scattered immunopositive cells mainly in interstitium. Immunohistochemical staining with mouse anti-human HGF antibody showed that HGF was distributed to the lung epithelial cells in IPF lung specimens obtained by open lung biopsy. These results suggest that HGF may play an important role in the pathogenesis of IPF. PMID- 9415351 TI - Diagnostic value of ferritin, haptoglobin, alpha 1-antitrypsin, lactate dehydrogenase and complement factors C3 and C4 in pleural effusion differentiation. AB - Searching to define diagnostic criteria for malignant and non-malignant pleural effusions, the differential diagnostic value of ferritin (FRT), haptoglobin (Hp), alpha 1-antitrypsin (alpha 1-AT), lactate dehydrogenase (LDH) and complement factors C3 and C4 were investigated prospectively in 100 consecutive patients with pleural effusions of various aetiologies. Pleural effusion FRT, C3 and C4 concentrations were found to be useful in differentiating exudates from transudates, so that transudates practically could be excluded in pleural effusion: serum FRT ratio lower than 0.5 and/or in pleural effusion values for C3 and C4 higher than 300 mg dl-1 and 70 mg dl-1, respectively. A pleural effusion: serum C3 ratio greater than 2 is seen only in malignant effusions. No discriminative pleural: serum ratio could be found in FRT and C4 values capable of differentiating malignant from non-malignant effusions. Pleural effusion alpha 1-AT and LDH values were elevated in exudates, as compared with transudates, and had an excellent sensitivity and predictive value, but low specificity, in differentiating malignant from non-malignant effusions. Finally, the sensitivity, specificity and positive predictive value of pleural effusion Hp concentrations were lower than those of FRT and complement factors C3 and C4, respectively. PMID- 9415352 TI - Attendance at an asthma educational intervention: characteristics of participants and non-participants. AB - As part of an evaluation of the patient education component of the Australian Asthma Management plan, a randomized, controlled trial of asthma education was conducted in 1994/95 at the outpatient asthma and allergy clinic of The Alfred Hospital, a tertiary referral hospital in Melbourne, Australia. The objective of the study was to investigate which demographic and clinical characteristics were associated with attendance at asthma educational session. A total of 125 asthmatics aged over 16 years agreed to participate in the programme, and full compliance with the programme was 43.2%. Allocation to immediate, rather than delayed, education and age were the only significant predictors of attendance. Subjects randomized to the intervention were approximately three times more likely to attend than control subjects (OR = 3.3, 95% CI 1.5-7.3). Asthmatics over 60 years old were approximately six times more likely to attend (OR = 6.6, 95% CI 2.2-19.8) than the age group 16-30 years. The increasing trend in attendance across age categories was highly significant (P < 0.001). There was no relationship between attendance and gender, medication, atopy, smoking status or the physical accessibility of the hospital. Despite offering incentives and conducting the education sessions at subjects' preferred times, their compliance in attending sessions was poor. Over half of the asthmatics, who had expressed interest, failed to attend for their educational sessions. An alternative strategy is required to improve participation by young and employed asthmatics at hospital-based asthma education programmes. PMID- 9415353 TI - A new bronchoscopic technique for the diagnosis of bacterial pneumonia in HIV positive patients. AB - The aim of the present study was to evaluate in HIV-positive patients with bacterial pneumonia, the diagnostic value of a new endoscopic technique that uses a single catheter to perform a telescopic plugged catheter (TPC) followed by a modified protected bronchoalveolar lavage (mpBAL). Fifty-eight HIV-positive patients with respiratory infection were included in the study. Samples from TPC and mpBAL were cultured quantitatively. Standard bronchoalveolar lavage was performed to rule out opportunistic infections. According to the clinical and microbiological results, patients were classified in the study group (27 with bacterial pneumonia) or the control group (31 without bacterial pneumonia). Sensitivity of TPC was 56% [95% confidence intervals (CI) 37-75%] and its specificity was 100%; these figures were 56% (CI, 37-75%) and 94% (CI, 86-100%) for mpBAL. When both techniques were assessed together, sensitivity increased to 70% (CI, 53-87%). The use of a single catheter reduced the cost of the originally described pBAL procedure by approximately 50%. The use of a single catheter to perform a TPC followed by a mpBAL can improve the diagnostic yield in HIV positive patients with bacterial pneumonia, and reduces its cost. PMID- 9415354 TI - Partial pulmonary sympathetic denervation by thoracoscopic D2-D3 sympathicolysis for essential hyperhidrosis: effect on the pulmonary diffusion capacity. AB - In patients with essential hyperhidrosis (EH), a pathological condition characterized by increased activity of the upper dorsal sympathetic ganglia D2 D3, anatomical interruption at the D2-D3 level by thoracoscopic sympathicolysis (TS) is a safe and effective treatment. The D2 and D3 ganglia, however, are also in the pathway of sympathetic lung innervation, which may influence the pulmonary diffusion capacity for carbon monoxide (expressed as transfer factor for CO:TLCO, and as transfer coefficient for CO:KCO). We therefore studied the effect of TS on TLCO and KCO in 50 EH patients: compared with pre-operative values, both TLCO ( 6.7%, P < 0.001) and KCO (-4.2%, P = 0.002) were significantly decreased at 6 weeks after bilateral TS, an effect which was independent of the smoking status of the patients. In order to explain this phenomenon, the following pharmacological interventions were studied: (1) oral beta 1 + 2-adrenoreceptor blockade with propranolol caused a comparable decrease of TLCO (-6.3%) and KCO ( 7.5%) in matched normal subjects, but had no effect on TLCO and KCO in EH patients prior to TS; and (2) subsequent inhalation of the beta 2-adrenoreceptor agonist salbutamol in a dosage suspected to cause alveolar beta-receptor stimulation had no effect on TLCO and KCO, neither in the normal subjects, nor in EH patients (before and after TS). Although the exact mechanism of the TS-induced decrease in TLCO and KCO remains speculative, these findings suggest that they may be related to a beta 1-adrenoreceptor-mediated change in pulmonary capillary membrane permeability, although TS-induced changes in pulmonary blood flow or an interplay of both mechanisms cannot be excluded. PMID- 9415355 TI - Tuberculosis in health service employees in Northern Ireland. AB - Tuberculosis can be transmitted from patients to health care workers. However, where the incidence of tuberculosis is low, and good infection control practices exist, the risk of health care workers acquiring the disease is likely to be small. The objective of this study was to determine the rate of notification of tuberculosis in health care workers in Northern Ireland compared with the general population. Information from the statutory tuberculosis notification forms for the period 1982-1991 was entered on to a computer database. Those patients involved in health care occupations were identified and age and sex standardized incidence rates were calculated. The overall notification rate for tuberculosis was 7.4 cases per 100,000 of general population. There was no significant increase in notification of tuberculosis among health care workers [standardized incidence ratio: 126% (95% CI 91-170)]. No cases were diagnosed as a result of screening methods performed during employment. It was concluded that health care workers in Northern Ireland did not have a significantly increased incidence of tuberculosis compared with the general population over the 10-year period studied. This suggests that the risk of transmission from patients to health care workers is negligible in the setting of a low general incidence of tuberculosis and good infection control practice. Under these circumstances, the present findings support the cessation of routine screening of health care workers. PMID- 9415356 TI - Impairment of muscle energy metabolism in patients with sleep apnoea syndrome. AB - Impairment of muscle energy metabolism has been demonstrated in normal subjects with chronic hypoxaemia (altitude chronic respiratory failure). The purpose of this study was to verify the hypothesis that a comparable condition could develop in patients with sleep apnoea syndrome (SAS), considering that they are exposed to prolonged and repeated hypoxaemia periods. Muscle metabolism was assessed in 11 patients with SAS performing a maximal effort on cycloergometer. In comparison with normal subjects, SAS patients reached lower maximal loads [144 +/- 7 vs. 182 +/- 10 W (P < 0.005)] and lower peak oxygen uptakes [26.4 +/- 1.2 vs 33.2 +/- 1.4 ml kg-1 min-1 (P < 0.005)]. Abnormal metabolic features were found: maximal blood lactate concentration was significantly lower than in normal subjects [0.034 +/- 0.004 vs. 0.044 +/- 0.002 mmol l-1 W-1 (P < 0.05)]; and lactate elimination rate, calculated during a 30-min recovery period, was reduced [0.127 +/- 0.017 vs, 0.175 +/- 0.014 mmol l-1 min-1 (P < 0.025)]. The extent of these anomalies correlated with the severity of SAS. The patients also showed higher maximal diastolic blood pressures than normal subjects [104 +/- 5 vs. 92 +/- 4 mmHg (P < 0.05)]. These results can be interpreted as indications of an impairment of muscle energy metabolism in patients with SAS. Decrease in maximum blood lactate concentration suggests an impairment of glycolytic metabolism, while decrease in the rate of lactate elimination indicates a defect in oxidative metabolism. Since no respiratory pathology apart from SAS was found in this group of patients, it seems legitimate to link the genesis of these impairments to repeated bouts of nocturnal hypoxaemia. PMID- 9415357 TI - Comparative studies of circulating KL-6, type III procollagen N-terminal peptide and type IV collagen 7S in patients with interstitial pneumonitis and alveolar pneumonia. AB - KL-6, a MUC1 mucin preferentially expressed in regenerating type 2 pneumocytes, has been reported to be a sensitive serum marker for evaluating the disease activity of interstitial pneumonitis (IP). Type III procollagen N-terminal peptide (PIIIP) and type IV collagen 7S (7S collagen) have also been reported to be useful in the serological evaluation of the activity. Their levels were measured and their serodiagnostic values were compared simultaneously in patients with IP and alveolar pneumonia. The study population was 45 patients with IP and 12 patients with alveolar pneumonia. Serum KL-6 levels were measured by a specific enzyme immunoassay, and both serum PIIIP and 7S collagen concentrations by their correspondent radioimmunoassay kits. There were no significant difference of serum C-reactive protein level, which was evaluated as an indicator of inflammatory process, between IP and alveolar pneumonia patients. In IP, the abnormally elevated rate of KL-6 [80% (36/45)] was significantly higher than those of PIIIP [40% (18/45)] and 7S collagen [40% (18/45)]. In alveolar pneumonia, the rate of KL-6 [0% (0/12)] was significantly lower than those of PIIIP [33% (4/12)] and 7S collagen [25% (3/12)]. There were no significant correlations among serum levels of the markers. These observations indicate that the serodiagnostic value of KL-6 for IP is superior to that of PIIIP and 7S collagen, and that KL-6 has a characteristic to discriminate IP from alveolar pneumonia. PMID- 9415358 TI - Left atrial tumour mimicking pulmonary embolism. AB - Primary cardiac tumours have been described as great imitators. They are rare, and clinical presentations are diverse. Diagnosis is usually made by two dimensional echocardiography. The present case report describes a case where a left atrial fibrosarcoma eluded diagnosis by echocardiography, and was eventually demonstrated by computed tomography. Management was complicated by the presence of persistent mismatch demonstrated by ventilation-perfusion lung scans. The likely mechanism underlying this phenomenon is discussed. PMID- 9415359 TI - Chemotherapy-induced myocardial necrosis in a patient with chronic lymphocytic leukemia. AB - Cardiac toxicity following the administration of chemotherapeutic agents is well documented. Vinca alkaloids, as well as high-dose cyclophosphamide, have been associated with myocardial ischemia. The present report describes a case of acute myocardial infarction occurring in a patient with no antecedent cardiac history who received both vincristine and conventional chemotherapeutic doses of cyclophosphamide for the treatment of chronic lymphocytic leukemia. Physicians should possess a heightened awareness of this potentially serious complication. PMID- 9415360 TI - Primary biphasic synovial sarcoma of the pleura. AB - Synovial sarcoma most commonly occurs in the peri-articular regions of the extremities. The present report describes a rare case of primary biphasic synovial sarcoma of the pleura in an 18-year-old female. The diagnosis was made on the basis of light microscopy, immunohistochemistry, electron microscopy and the characteristic translocation found on cytogenetic analysis. Synovial sarcoma should be included in the differential diagnoses of pleural tumors. PMID- 9415361 TI - Non-Hodgkin's lymphoma presenting as Pancoast's syndrome. AB - Pancoast's syndrome is generally caused by primary or metastatic epithelial tumors. Other causes of the syndrome are unusual but well described. The present case report describes a rare case of Pancoast's syndrome caused by non-Hodgkin's lymphoma. This report emphasises the importance of establishing a firm pathologic diagnosis of the etiology of Pancoast's syndrome before instituting treatment. PMID- 9415363 TI - Filtering effect of cone oil droplets detected in the P-III response spectra of Japanese quail. AB - While absorption spectra of bird cone visual pigments have been well studied, physiological study of bird cone cells has been less advanced owing to their small sizes. We measured the P-III components of electroretinograms (ERG) from isolated retinas of Japanese quail. We recorded responses to monochromatic flashes of equal photon numbers, and found that the shape of the response spectrum is dependent on the incident direction of the flashes. The spectrum obtained with the flashes from the cornea side had a steeper peak around 500 nm than that with flashes from the receptor side. This is clear electrophysiological evidence of the filtering effect of the oil droplets in the cone cells, which has long been suspected. We analyzed these spectra with respect to the absorption spectra of cone pigments and transmittance spectra of oil droplets. PMID- 9415362 TI - Rod inputs to macaque ganglion cells. AB - The strength of rod inputs to ganglion cells was assessed in the macaque retina at retinal positions within 3-15 deg eccentricity. The experimental paradigm used temporally modulated heterochromatic lights whose relative phase was varied. This paradigm provided a sensitive test to detect rod input. In parvocellular (PC) pathway cells, the gain of the cone-driven signal decreased with decrease in luminance. At 2 td a weak rod response, of a few impulses per second for 100% rod modulation, was revealed in about 60% of cells. For blue-on cells, the cone driven response also decreased with retinal illuminance, but no rod response could be found. In magnocellular (MC) pathway cells, rod input was much more apparent. Responses became rod dominated at and below 20 td; we cannot exclude rod intrusion at higher retinal illuminances. Responsivity was maintained even at low retinal illuminances. Temporal-frequency dependent rod-cone interactions were observed in MC-pathway cells. Rod responses were of longer latency than cone responses, but there was no evidence of any difference in rod latency between parvocellular and magnocellular pathways. PMID- 9415364 TI - Curvilinearity, covariance, and regularity in perceptual groups. AB - A curvilinear pattern among a series of visual items (e.g., dots) can be regarded as a kind of probabilistic inference, in which each consecutive angle, regarded independently, is more nearly collinear than would be expected by chance alone. This paper investigates judgments of curvilinearity as a function of the joint distribution of successive inter-dot angles. Subjects were asked to classify 4- and 5-dot configurations as having been generated by a curvilinear generating process, vs independently. Their results are distributed as a gaussian over the inter-dot angles with mean 0 deg (collinear), with a negative correlation between successive angles, but negligible correlation between non-successive angles. This suggests that curvilinearity is evaluated in a 4-dot window moving along the chain of dots, evaluating collinearity and smoothness but ignoring higher-order relationships. Moreover, the probabilistic model provides a remarkably precise numeric prediction of the magnitude of the correlation. Subjects also showed a reliable preference for equal spacing of dots along the virtual curve. PMID- 9415365 TI - Similar mechanisms underlie simultaneous brightness contrast and grating induction. AB - The experiments explore whether the mechanism(s) underlying grating induction (GI) can also account for simultaneous brightness contrast (SBC). At each of three test field heights (1, 3 and 6 deg), point-by-point brightness matches were obtained from two subjects for test field widths of 32 deg (GI condition), 14, 12, 8, 6, 3 and 1 deg. The point-by-point brightness matches were quantitatively compared, using GI condition matches as a standard, to assess systematic alterations in the structure and average magnitude of brightness and darkness induction within the test fields as a function of changing test field height and width. In the wider test fields induction structure was present and was generally well-accounted for by the GI condition sinewave predictions. As test field width decreased the sinewave amplitude of the induced structure in the test field decreased (i.e., flattened), and eventually became negative (i.e., showed a reverse cusping) at the narrower test field widths. As expected, both subjects showed a decrease in overall levels of brightness and darkness induction with increasing test field height. For any particular test field height, however, relative brightness increased with decreasing test field width. This brightness increase began at larger test field widths as test field height increased. The results are parsimoniously accounted for by the output of a weighted, octave interval array of seven difference-of-gaussian filters. This array of filters differs from those previously employed to model various aspects of spatial vision in that it includes filters tuned to much lower spatial frequencies. The two dimensional output of this same array of filters also accounts for the GI demonstrations of Zaidi [(1989) Vision Research, 29, 691-697], Shapley and Reid's [(1985) Proceedings of the National Academy of Sciences USA, 82, 5983-5986] contrast and assimilation demonstration, and the induced spots seen at the street intersections of the Hermann Grid. The physiological plausibility of the filter array explanation of brightness induction is discussed, along with a consideration of its relationship to other models of brightness perception. PMID- 9415366 TI - Spatial limitation of vertical-size disparity processing. AB - We investigated the upper limit of horizontal spatial modulation of vertical-size disparity in a textured surface for the perception of depth. In Experiment 1 subjects matched the appearance of a surface with modulated horizontal-size disparity to that of a surface with modulated vertical-size disparity. In Experiment 2 we determined the threshold amplitude of modulation of vertical-size disparity required for the perception of depth as a function of the spatial frequency of disparity modulation. The results indicate that sensations of depth are not elicited by modulations of vertical-size disparity of any amplitude at spatial frequencies higher than about 0.04 c/deg. We conclude that vertical disparities are averaged within about 20 deg-wide areas and suggest that this global measurement is used to scale local horizontal disparities for the perception of surface slant. PMID- 9415367 TI - Velocity decomposition and surface decomposition--reciprocal interactions between motion and form processing. AB - Physically unidirectional motion of short-lived random dot arrays was found to perceptually decompose into two motion components (velocity decomposition) in a configuration in which two squares appear to partially overlap transparently (surface decomposition). In the experiments in which the velocity of the short lived random dots in the overlapping area was varied, both the velocity decomposition and the surface decomposition were found to be strongest when the velocity of the overlapping area was close to the vector sum of the velocities of random dots in adjacent non-overlapping areas. On the other hand, neither velocity decomposition nor surface decomposition was found either when random dot arrays were put in occlusion configurations or when continuous random dots were used. While previous studies have indicated a one-way influence either from motion to form processing, or from form to motion processing, the present study further suggests that there is a strong reciprocal interaction between motion and form processing. A possibility is that the reciprocal interaction is iterative so that the representations for velocity and surface decomposition are gradually formed. PMID- 9415368 TI - The influence of subject instruction on horizontal and vertical vergence tracking. AB - Previously it has been reported that horizontal disparity vergence is strongly influenced by subject instructions to vary attention or tracking effort. This paper describes experiments which compared these instruction effects on horizontal and vertical disparity vergence. Within-trial comparisons were made possible by use of oblique (combined horizontal and vertical) disparity modulation. Subjects viewed a flat, fully correlated, dynamic random noise stereogram pattern through stationary circular apertures, with a small stationary fixation cross superimposed in the center. The disparity of the noise pattern was either modulated sinusoidally or changed abruptly. Subjects were instructed either to (1) hold fixation on the cross and ignore the disparity modulation of the noise pattern; or (2) follow the movement of the noise pattern as accurately as possible. Subjects showed clear effects of instruction on the horizontal component of tracking, but showed little or no effect on the vertical component. Horizontal and vertical components of oblique vergence tracking appear to be largely independent, and vertical vergence is affected minimally, if at all, by an effort to track. PMID- 9415369 TI - Fillers and spaces in text: the importance of word recognition during reading. AB - Current theories of reading eye movements claim that reading saccades are programmed primarily on the basis of information about the length of the upcoming word, determined by low-level visual processes that detect spaces to the right of fixation. Many studies attempted to test this claim by filling spaces between words with various non-space symbols (fillers). This manipulation, however, confounds the effect of inserting extraneous characters into text with the effect of obscuring word boundaries by filling spaces. We performed the control conditions necessary to unconfound these effects. Skilled readers read continuous stories aloud and silently. Three factors were varied: (i) position of the fillers in the text (at the beginning, the end, or surrounding each word); (ii) the presence or absence of spaces in the text; and (iii) the effect of the type of filler on word recognition (from greatest effect to least effect: Latin letters, Greek letters, digits and shaded boxes). The effect of fillers on reading depended more on the type of filler than on the presence of spaces. The greater effect the fillers had on word recognition, the more they showed reading. Surrounding each word with digits or Greek letters slowed reading as much as filling spaces with these symbols. Surrounding each word with randomly chosen letters, while preserving spaces, slowed reading by 44-75%--as much as, or more than, removing spaces from normal text. Removing spaces from text with Latin letter fillers slowed reading by only 10-20% more. We conclude that fillers in text disrupt reading by affecting word recognition directly, without necessarily affecting the eye movement pattern. PMID- 9415370 TI - The spatial mechanisms mediating symmetry perception. AB - This paper examines the role of spatial frequency and orientation tuned channels in the perception of visual symmetry. Subjects discriminated between band-pass filtered, white noise textures that either did or did not contain vertical bilateral symmetry (VBS, i.e., around a vertical midline) as a function of the spatial phase disruption imposed on the images. Resistance to phase noise is largely scale-invariant for isotropically filtered images, but horizontally filtered images are consistently more noise-resistant than vertical. However, when stimuli are rotated through 90 deg (horizontal bilateral symmetry, HBS) performance is better with vertically filtered images suggesting a general advantage for orientations orthogonal to the axis of symmetry. At these orientations symmetry may be signaled directly by clusters of features along the axis. Our data further suggest that the established disadvantage for HBS may be attributable to an over-reliance on the output of horizontal filters. We compare models which exploit feature clustering around the axis by measuring the co alignment in the output of oriented filters. Models using filters oriented orthogonal to the axis of symmetry predict the psychophysical performance for isotropic patterns and for patterns filtered orthogonal to the axis. For patterns filtered parallel to the axis, our data suggest that visual attention may play a role. PMID- 9415371 TI - Hidden visual capabilities in mentally retarded subjects diagnosed as deaf-blind. AB - The visual acuity of twelve multi-handicapped, mentally retarded subjects, diagnosed as deaf-blind, was measured on two occasions with the Teller Acuity Cards (TAC). Eight subjects scored above the criterion for legally blind and the results of six of these indicated various degrees of poor to approaching-normal eyesight. To evaluate high-level vision four subjects were tested with the Fagan Test, assessing visual recognition memory for faces subsequent to familiarization with the preferential looking technique. The results for three subjects showed evidence for perceptual recognition. It is concluded that TAC combined with the Fagan Test may detect perceptual capacities unnoticed by clinical observation. PMID- 9415372 TI - Episodic acute hypotonia after Nd:YAG laser capsulotomy--retinal function and choroidal swelling. AB - In a patient with uveitis who had been treated with Nd:YAG laser capsulotomy after cataract surgery, several episodes of acute hypotonia occurred which were associated with changes in clinical tests of the eye and of visual function. Immunosuppressive and immunomodulating treatment appeared to reverse the changes in intraocular pressure and normalise the test results. The significance of these observations is discussed. PMID- 9415373 TI - A synergy of technologies: combining laser microsurgery with green fluorescent protein tagging. AB - When focused through an objective lens with a high numerical aperture, nanosecond pulses of high-intensity green (532-nm) laser light can be used to selectively destroy any cellular component whose boundaries can be defined by light microscopy. These components include, for example, chromosomes, spindle fibers, bundles of keratin, or actin filaments, mitochondria, vacuoles, and so forth. In addition, the definition of poorly resolved components can be enhanced for selective destruction by tagging one or more of their constituent proteins with green fluorescence protein (GFP). As a example we show that the centrosome in living PtK1 cells can be clearly defined, and then destroyed by green laser light, after transforming the cells with gamma-tubulin/GFP fusion protein. In some transformed cells it is even possible to target and selectively destroy just one of the centrioles. PMID- 9415374 TI - Translocation of myelin basic protein mRNA in oligodendrocytes requires microtubules and kinesin. AB - Myelin basic protein (MBP) mRNA is localized to the myelin membranes of oligodendrocytes. When exogenous MBP mRNA is microinjected into oligodendrocytes in culture, it is transported along the processes and localized to the myelin compartment in a multistep intracellular RNA trafficking pathway. In the work described here, oligodendrocytes were treated with agents that affect the cytoskeleton including: nocodazole, to disrupt microtubules; taxol, to stabilize microtubules; cytochalasin, to disrupt microfilaments; and kinesin anti-sense oligonucleotide, to suppress kinesin expression. Digoxigenin-labeled MBP mRNA was microinjected into the treated cells and the extent of translocation of the microinjected RNA was determined by confocal microscopy. Nocodazole, taxol, and kinesin anti-sense oligonucleotide inhibited translocation of microinjected MBP mRNA, while cytochalasin B and kinesin sense oligonucleotide did not. These results indicate that translocation of MBP mRNA in oligodendrocytes requires intact microtubules and kinesin but does not require intact microfilaments. The results are discussed in relation to the current multistep model for intracellular RNA trafficking in oligodendrocytes. PMID- 9415375 TI - Cold-adapted microtubules: characterization of tubulin posttranslational modifications in the Antarctic ciliate Euplotes focardii. AB - In cold poikilotherm organisms, microtubule assembly is promoted at temperatures below 4 degrees C and cold-induced depolymerization is prevented. On the basis of the results of investigations on cold-adapted fishes, the property of cold adaptation is ascribed to intrinsic characteristics of the tubulins. To fully understand cold adaptation, we studied the tubulins of Euplotes focardii, an Antarctic ciliated protozoan adapted to temperatures ranging from -2 to +4 degrees C. In this organism, we had previously sequenced one beta-tubulin gene and, then identified three other genes (denoted as beta-T1, beta-T2, beta-T3 and beta-T4). Here we report that the amino acid sequence of the carboxy-terminal domain predicted from the beta-T3 gene (apparently the most expressed of the gene family) contains six modifications (five substitutions and one insertion) of conserved residues, unique with respect to all the other known beta-tubulin sequences. These modifications can change the structural conformation of the carboxy-terminal domain. Furthermore, in the variable terminal end of that domain, a consensus sequence for a phosphorylation site is present, and the residue Glu-438, the most frequent site for polyglutamylation in beta-tubulin, is substituted by Asp. Starting from these observations, we showed that in E. focardii only alpha-tubulin is polyglutamylated, while beta-tubulin undergoes phosphorylation. Polyglutamylated microtubules appear to colocalize with cilia and microtubular bundles, all structures in which microtubules undergo a sliding process. This finding supports the idea that alpha-tubulin polyglutamylation is involved in the interaction between tubulin and motor microtubule-associated proteins. Phosphorylation, usually a rare posttranslational modification of beta tubulin, which is found extensively distributed in the beta-tubulin of this cold adapted organism, may play a determinant role in the dynamic of polymerization and depolymerization at low temperatures. PMID- 9415376 TI - Actin-filament motion in the in vitro motility assay has a periodic component. AB - The interaction between actin and myosin can be studied in the in vitro motility assay, where fluorescently labelled actin filaments are observed to move over a lawn of myosin heads. To examine details of this movement, we measured systematically the velocities of the front end, rear end, and centroid of the actin filament as the filament translated over the assay surface. We found that these velocities exhibited an unexpectedly periodic component, alternating regularly between high and low values, superimposed on the steady velocity component. The period of the oscillatory component was approximately 380 ms. When translation was stopped by an increase in osmolarity, the filaments wiggled with a periodicity similar to the translating filament, implying that wiggling and translation may be related. Rigor filaments showed no periodicity. From the frequency content of the auto- and cross-correlation functions derived from the velocities of the front end, rear end, and centroid of the actin filament, we infer a deterministic, possibly wave-like process travelling along the actin filament. Potential molecular mechanisms underlying this phenomenon are considered. PMID- 9415377 TI - Role of cadherins in the transendothelial migration of melanoma cells in culture. AB - Transmigration of cancer cells through the vascular endothelium (diapedesis) is a key event in tumor metastasis. To investigate mechanisms involved in diapedesis, we used laser scanning confocal microscopy to examine the distribution of cadherins of WM239 melanoma cells as they migrated through a monolayer of activated human umbilical vein endothelial cells (EC) cultured on matrigel. Cadherins, including VE-cadherin, but not N-cadherin, were enriched in contacts between EC, whereas N-cadherin, but not VE-cadherin, was found in contacts between melanoma cells. During the early stages of diapedesis, EC located below the attached melanoma cells decreased in height and VE-cadherin disappeared from the EC contact located underneath the melanoma cell. Transendothelial migration began with small melanoma cell processes penetrating the VE-cadherin-negative regions between the EC. Subsequently, melanoma cells became intercalated between EC. Despite the absence of both VE-cadherin and N-cadherin, other members of the cadherin family were present in the heterotypic contacts between EC and melanoma cells. EC surrounding the intercalated melanoma cell subsequently extended processes and spread over the melanoma cell to re-form the endothelial monolayer. Interestingly, the leading margins of these EC processes contained high levels of N-cadherin, but not VE-cadherin. VE-cadherin-rich cell-cell contacts, however, reformed between advancing endothelial processes when they met above the melanoma cell. As the melanoma cells came into contact with the underlying matrigel, they spread out and adopted a fibroblast-like morphology. Addition of anti-N-cadherin antibodies to the assay resulted in a delay in the transendothelial migration of melanoma cells. Together, these results suggest that EC actively participate in diapedesis by disassembling and reassembling VE-cadherin-rich adherens junctions, and that N-cadherin plays an important role in the transmigration of melanoma cells and the reclosure of the endothelium. PMID- 9415378 TI - Jasplakinolide, a novel actin targeting peptide, inhibits cell growth and induces actin filament polymerization in the green alga Micrasterias. AB - Jasplakinolide, a naturally occurring cyclodepsipeptide from the marine sponge Jaspis sp., known to induce actin polymerization and stabilization in vitro, markedly influences the morphogenetic process in the green alga Micrasterias when used in concentrations higher than 3 microM. Development of Micrasterias is inhibited or strongly retarded, malformations occur, and large vacuoles are formed. At the ultrastructural level, dense abnormal accumulations of filamentous structures have been found indicating actin filament polymerizing activities of the drug in situ. Moreover, displacement of organelles and aggregations of endoplasmic reticulum (ER) cisternae have been observed. Microtubule (MT) arrangement and MT-dependent processes remain undisturbed. Cells allowed to recover from jasplakinolide treatment continue their growth but show severe changes in the cell pattern and displacement of organelles, suggesting that even after removal of the drug, some basic features for the morphogenetic process remain altered. Jasplakinolide might be a useful tool for investigations on actin dependent processes in the future. PMID- 9415379 TI - Relationship of actin, microtubules, and crosswall synthesis during septation in Aspergillus nidulans. AB - Studies of cytokinesis in animal cells demonstrate that microtubules play an important role in signaling the position of the actin-containing contractile ring and subsequent formation of the cleavage furrow. Septation in several fungi closely resembles animal cell cytokinesis in that a circumferential ring of actin is visible at the incipient division site. However, this does not necessarily mean that division is contractile since actin may also serve to localize septal wall synthesis. In addition, several studies in fission yeast have suggested that microtubules are dispensable for actin ring formation. We have used synchronized cells and fluorescence microscopy to follow actin structures, nuclear division and septal wall synthesis during septation in Aspergillus nidulans. Our data suggest that actin first appears at the septum site as a circumferential ring and that it later broadens and invaginates, forming an hourglass-shaped structure coincident with septal cell wall synthesis. Depolymerization of microtubules early in septation prevents circumferential actin ring formation. Depolymerization of microtubules after circumferential actin ring formation blocks both the progression to invaginating bands and septal wall synthesis. In contrast to studies in yeast cells, our data suggest that microtubules are required for both the initiation and progression of septation in A. nidulans. PMID- 9415380 TI - Type II myosin involved in cytokinesis in the fission yeast, Schizosaccharomyces pombe. AB - We have cloned an unique gene encoding the heavy chain of a type II myosin in the fission yeast, Schizosaccharomyces pombe. The myo2+ gene encodes a protein of 1526 amino acids with a predicted molecular weight of 177 kDa and containing consensus binding motifs for both essential and regulatory light chains. The S. pombe myo2+ head domain is 45% identical to myosin IIs from Saccharomyces cerevisiae and Homo sapiens and 40% identical to Drosophila melanogaster Structurally, myo2+ most closely resembles budding yeast MYO1, the tails of both myosin IIs containing a number of proline residues that are predicted to substantially disrupt the ability of these myosins to form coiled coils. The myo2+ gene is located on chromosome III, 8.3 map units from ade6+. Deletion of approximately 70% of the coding sequence of myo2+ is lethal but myo2delta spores can acquire a suppressor mutation that allows them to form viable microcolonies consisting of filaments of branched cells with aberrant septa. Overexpression of myo2+ results in the inhibition of cytokinesis; cells become elongated and multinucleate and fail to assemble a functional cytokinetic actin ring and are either aseptate or form aberrant septa. These results suggest that a contractile actin-myosin based cytokinetic mechanism appeared early in the evolution of eukaryotic cells and further emphasise the utility of fission yeast as a model organism in which to study the molecular and cellular basis of cytokinesis. PMID- 9415381 TI - Evidence for several roles of dynein in pigment transport in melanophores. AB - Melanophores are specialized cells that transport pigment granules to and from the cell center, giving animals the ability to change skin color. A kinesin related plus-end motor has previously been shown to be responsible for pigment granule dispersion [V.I. Rodionov, F.K. Gyoeva, and V.I. Gelfand. Proc. Natl. Acad. Sci. USA. 1991, 88:4956-4960]. Here, we have microinjected a dynein antibody (70.1) into cultured cod (Gadus morhua) melanophores and used the dynein inhibitor vanadate on permeabilized melanophores in skin pieces, to examine the role of the microtubule minus-end motor dynein in these cells. Both pigment granule aggregation and maintenance of the spherical central pigment mass (CPM) were inhibited by the antibody and by vanadate. Vanadate or antibody treatment of cells with aggregated pigment did not induce pigment dispersion. However, when the antibody-injected cells were induced to disperse pigment, the pigment moved farther to the cell periphery, which resulted in a depletion of pigment in the cell center. Similar superdispersion of previously uniformly distributed pigment was also seen when the antibody was injected in melanophores with dispersed pigment. Our results demonstrate that both pigment aggregation and maintenance of the CPM are dynein-dependent processes. Our data further show that dynein is involved in the homogeneous distribution of dispersed pigment. These results suggest that both dynein and kinesin are active in keeping pigment granules dispersed throughout the cytoplasm, transporting pigment granules in opposite directions. The possibility that dynein is continuously active during both aggregation and dispersion, while kinesin might be the target for regulation, is discussed. PMID- 9415383 TI - Vagus nerve stimulation for intractable epilepsy: a review. AB - Electrical stimulation of the vagus nerve in the neck by using a programmable stimulator similar to a cardiac pacemaker is being explored as a treatment for epilepsy. There is sound rationale based on studies of animal seizure models for pursuing this treatment modality, and early clinical trials provide support for efficacy in patients with intractable epilepsy at least equivalent to that of some of the new antiepileptic drugs. Safety and tolerability have been demonstrated in >800 patients worldwide since the first implant in 1988. Most of these had partial seizures for which resective epilepsy surgery was not feasible or had failed, but efficacy of vagal stimulation appears to be the same for both partial and generalized epilepsy. Specific selection criteria for this procedure have yet to be established, and further studies are warranted to determine whether vagal stimulation becomes an accepted procedure for epilepsy management. PMID- 9415384 TI - Diaphragm pacing for respiratory insufficiency. AB - Diaphragm pacing (DP) by electrical stimulation of the phrenic nerve offers important advantages to a highly select group of patients with respiratory paralysis. The patient wears an external radiofrequency (RF) transmitter over an implanted receiver, and a stimulating current is induced without the need for any transcutaneous wires. We review the conditions and requirements of patients who may benefit most from DP. We outline the preoperative evaluation and procedures for surgical implantation. We discuss the risk of diaphragmatic fatigue posed by initiation of DP and the use of gradual conditioning to limit this problem. Other problems encountered by patients in the course of DP can be minimized by well instructed home caregivers and by systematic medical follow-up. Although a few patients derive considerable benefit from DP, many patients with respiratory paralysis are better treated by less invasive means such as nasal bilevel positive airway pressure or intermittent positive pressure ventilation, which we also review. PMID- 9415385 TI - Peripheral nerve stimulation for restoration of motor function. AB - This review paper discusses the use of electrical stimulation to restore function after upper motor neurone type of paralysis. It describes the basic physiology of electrical stimulation, the electrophysiology and biomaterials associated with using metal electrodes to deliver charge to living tissue, and also the adverse effects of stimulation. The central concepts of electrode applications, stimulus parameters, muscle fatigue, and stimulation control are covered. Next, a survey of clinical applications is made with focus on upper and lower limb applications. A concluding section mentions the current status of commercial products available for stimulation. PMID- 9415386 TI - Event-related potentials associated with judgment: comparison of S1- and S2 choice conditions in a contingent negative variation (CNV) paradigm. AB - To elucidate the functional role of association cortex in judgment and decision making about external stimuli, the scalp topography of contingent negative variation (CNV) recorded in 9 normal volunteers was compared in two settings; dichotomous S1- and S2-choice paradigms using right wrist extension or flexion movement upon S2 as the choice reaction task with an S1-S2 interstimulus interval of 2 s. Two main findings were obtained. First, early CNV in the S1-choice paradigm consisted of a frontal to frontopolar midline negative potential, most likely representing judgment more than orienting response, and a left-sided parietal positive potential, at least partially representing P3b, whereas in the S2-choice paradigm it consisted only of a smaller frontal negative potential shift, most likely representing merely an orienting response. This suggests that the prefrontal and left parietal association cortexes might be active in the judgment and decision-making process. Second, after S2, two positive peaks (334 and 545 ms after S2) were observed at the midline central to left parietal areas only in the S2-choice paradigm. In the S1-choice paradigm, one large negative peak (275 ms after S2) was observed. We postulate that the first peak of the S2 choice paradigm is related to decision making or judgment and that the second peak is related to the following cognitive process. We conclude that at least the prefrontal and parietal association cortexes generate transient potentials accompanying judgment, and possibly decision making. PMID- 9415387 TI - Digital conversion of paper electroencephalograms using a hand scanner. AB - We report a method of quantifying the analog signal of paper-recorded EEGs that involves readily available technology, including a standard personal computer and a compatible hand scanner. Simulations assessed the effects of erroneously scanning the paper at a slight angle and estimating pen arc distortion; these effects were demonstrated to be insignificant. The method allows application of several quantitative techniques, including power spectral analysis and determination of the squared coherence between homologous regions. In a companion study, we applied this technique to compare the coherence in patients with alpha pattern coma to coherence in normal subjects. PMID- 9415388 TI - Comparison between the alpha pattern in normal subjects and in alpha pattern coma. AB - Alpha pattern coma (APC) is an uncommon clinical EEG pattern in comatose patients, most commonly in association with anoxic-ischemic encephalopathy after cardiac arrest. Despite the pattern's striking similarity to that of the normal awake EEG, there are theoretical and experimental reasons for believing that the two rhythms result from different processes. The analysis of quantitative differences in APC from normal rhythms requires computer analysis. Because most cases of this rare entity have been collected over the years on paper traces, computer analysis appears implausible. In a companion article, we describe a method to quantify sections of paper EEGs. We applied this method to EEGs of five APC patients and five normal controls and noted a significant difference in the coherence between the two hemispheres in the alpha range. This finding is in keeping with theoretical, experimental, and clinical observations suggesting that APC may result from significant thalamo-cortical disruption. PMID- 9415389 TI - Effects of handedness on movement-related changes of central beta rhythms. AB - The effects of handedness on the movement-related changes in beta rhythms (14-30 Hz) in the left and right perirolandic area were analyzed in 12 right-handed and 11 left-handed subjects. The motor task consisted of unilateral brisk or slow self-paced extension of the right or left index finger. The handedness effects were as follows. First, in both handedness groups, the premovement desynchronization of beta rhythms at both hemispheres was greatest before slow movement of the "nondominant" finger, especially at electrodes presumably overlying the MI areas. Second, the lefthanded group showed less desynchronization in both hemispheres during execution of a slow movement than the righthanded group. Third, the postmovement beta synchronization showed a contralateral preponderance which was greater after movements of the nondominant than the "dominant" finger in the righthanded group and was equal for both fingers in the lefthanded group. The results suggest that handedness effects on movement-related changes in central beta rhythms are coupled to movements of the nondominant finger and that their manifestation differs in the pre- and postmovement periods. PMID- 9415390 TI - Near- and far-fields: source characteristics and the conducting medium in neurophysiology. AB - It is possible to appreciate the production of far-field potentials by considering constant current dipolar source voltage distributions in bounded volumes, especially when they are stretched in one direction, e.g., a cylinder. An essentially nondeclining voltage is detected when the recording electrodes are on opposite sides of, and relatively far from, the dipolar source. This voltage maintains its (a) latency, (b) amplitude, (c) morphology, and (d) polarity even if recordings are performed a whole body length away. These four criteria define far-field potentials. A propagating action potential (AP) can be conceptualized as a linear quadrupole or the summation of two dipoles "back-to-back" (+ - - +). The far-field components of the summated dipoles cancel resulting in the anticipated triphasic waveform for APs with only near-field characteristics, not meeting the first three criteria above. Far-field potentials can be transiently generated when any propagating AP constitutes a net "real" or "virtual" dipolar source. "Real" dipolar sources can occur if an AP encounters the termination of excitable tissue, an alteration in conduction velocity, curvature in excitable tissue resulting in a change in propagation direction, or an abrupt change in resistance of the excitable tissue. Virtual dipolar sources may be produced if an AP encounters a change in the size or shape of the extracellular medium or a transition in extracellular conductivity. PMID- 9415391 TI - Dr. F. H. Lewey. PMID- 9415392 TI - How Dr. Lewy (Lewey) spelled his name. PMID- 9415393 TI - Physiological role of Akt in insulin-stimulated translocation of GLUT4 in transfected rat adipose cells. AB - Stimulation of glucose transport is among the most important metabolic actions of insulin. Studies in adipose cells have demonstrated that insulin stimulates its receptor to phosphorylate tyrosine residues in IRS-1, leading to activation of phosphatidylinositol 3-kinase, which plays a necessary role in mediating the translocation of the insulin-responsive glucose transporter GLUT4 to the cell surface. Akt is a serine-threonine kinase recently identified as a direct downstream target of phosphatidylinositol 3-kinase. A previous study in 3T3-L1 cells showed that overexpression of a constitutively active mutant of Akt is sufficient to recruit GLUT4 to the cell surface. Since effects of overexpression of signaling molecules in tissue culture models do not always reflect physiological function, we have overexpressed a dominant inhibitory mutant of Akt in rat adipose cells to investigate the effects of inhibiting endogenous Akt in a physiologically relevant insulin target cell. Cells were transfected with either wild type (Akt-WT), constitutively active (Akt-myr), or dominant inhibitory (Akt K179A) forms of Akt, and effects of overexpression of these constructs on insulin stimulated translocation of a cotransfected epitope-tagged GLUT4 were studied. Overexpression of Akt-WT resulted in significant translocation of GLUT4 to the cell surface even in the absence of insulin. Interestingly, overexpression of Akt myr resulted in an even larger effect that was independent of insulin. More importantly, overexpression of Akt-K179A (kinase-inactive mutant) significantly inhibited insulin-stimulated translocation of GLUT4. Taken together, our data suggest that Akt is not only capable of stimulating the translocation of GLUT4 but that endogenous Akt is likely to play a significant physiological role in insulin-stimulated glucose uptake in insulin targets such as muscle and adipose tissue. PMID- 9415394 TI - Modulation of Igf2 genomic imprinting in mice induced by 5-azacytidine, an inhibitor of DNA methylation. AB - The adjacent genes, insulin-like growth factor 2 (Igf2) and H19, are imprinted in both mouse and human. While Igf2 is expressed from the paternal allele, H19 is transcribed exclusively from the maternal allele. To explore the underlying mechanism of Igf2 and H19 imprinting, we studied the effect of DNA demethylation on allelic expression by injecting mice with the demethylating agent 5 azacytidine (5-aza-C). We observed a > or = 2-fold increase in the abundance of Igf2 mRNA in liver from treated mice compared with that of control mice. There was no significant change in Igf2 or H19 expression in brain. In the 5-aza-C treated mice, there was dramatic modulation of Igf2 imprinting. In some tissues, Igf2 was expressed biallelically, while in other tissues, the paternal allele was silenced and the normally imprinted maternal allele was expressed, an example of allelic switching. There was no change in the normal biallelic pattern of Igf2 expression in brain. H19, on the other hand, remained imprinted in all tissues in mice treated with 5-aza-C. These results provide the first example of a pharmacological manipulation of genomic imprinting of an endogenous gene in vivo and further implicate DNA methylation as an important factor in maintaining the differential allelic expression of the Igf2 gene. PMID- 9415395 TI - Early signaling events triggered by peroxovanadium [bpV(phen)] are insulin receptor kinase (IRK)-dependent: specificity of inhibition of IRK-associated protein tyrosine phosphatase(s) by bpV(phen). AB - Peroxovanadiums (pVs) are potent protein tyrosine phosphatase (PTP) inhibitors with insulin-mimetic properties in vivo and in vitro. We have established the existence of an insulin receptor kinase (IRK)-associated PTP whose inhibition by pVs correlates closely with IRK tyrosine phosphorylation, activation, and downstream signaling. pVs have also been shown to activate various tyrosine kinases (TKs) that could participate in activation of the insulin-signaling pathway. In the present study we have sought to determine whether pV-induced IRK tyrosine phosphorylation requires the intrinsic kinase activity of the IRK, and whether IRK activation is necessary to realize the early steps in the insulin signaling cascade. To address this we evaluated the effect of a pure pV compound, bis peroxovanadium 1,10-phenanthroline [bpV(phen)], in HTC rat hepatoma cells overexpressing normal (HTC-IR) or kinase-deficient (HTC-M1030) mutant IRKs. We showed that at a dose of 0.1 mM, but not 1 mM, bpV(phen) induced IRK-dependent events. Thus, 0.1 mM bpV(phen) increased tyrosine phosphorylation and IRK activity in HTC-IR but not HTC-M1030 cells. Tyrosine phosphorylation of insulin signal-transducing molecules was promoted in HTC-IR but not HTC-M1030 cells by bpV(phen). The association of p185 and p60 with the src homology-2 (SH2) domains of Syp and the p85-regulatory subunit of phosphatidylinositol 3'-kinase was induced by bpV(phen) in HTC-IR, but not in HTC-M1030 cells, as was insulin receptor substrate-1-associated phosphatidylinositol 3'-kinase activity. Thus autophosphorylation and activation of the IRK by bpV(phen) is effected by the IRK itself, and the early events in the insulin- signaling cascade follow from this activation event. This establishes a critical role for PTP(s) in the regulation of IRK activity. bpV(phen) could be distinguished from insulin only in its ability to activate ERK1 in HTC-M1030 cells, thus indicating that this event is IRK independent, consistent with our previous hypothesis that bpV(phen) inhibits a PTP involved in the negative regulation of mitogen-activated protein kinases. PMID- 9415396 TI - Interaction of wild type and dominant-negative p55PIK regulatory subunit of phosphatidylinositol 3-kinase with insulin-like growth factor-1 signaling proteins. AB - In a first series of experiments done in the yeast two-hybrid system, we investigated the nature of protein-protein interaction between the regulatory subunit of phosphatidylinositol 3-kinase (PI 3-kinase), p55PIK, and several of its potential signaling partners. The region between the Src homology 2 (SH2) domains of p55PIK bound to the NH2 terminus region of p110alpha, as previously shown for p85alpha. Moreover, we found that the insulin-like growth factor-1 receptor (IGF-IR) bound to p55PIK; the interaction occurred at the receptor tyrosine 1316 and involved both p55PIK SH2 domains. Interaction between p55PIK and IGF-IR was seen not only in the yeast two-hybrid system, but also using in vitro binding and coimmunoprecipitation of lysates from IGF-1 stimulated 293 cells overexpressing p55PIK. Further, IGF-I stimulation of these cells led to tyrosine phosphorylation of p55PIK. In 293 cells association of p55PIK with insulin receptor substrate-1 and with IGF-IR was dependent on PI 3-kinase, since it was increased by wortmannin, an inhibitor of PI 3-kinase. Further, by deleting amino acids 203-217 of p55PIK inter-SH2 domain, we engineered a p55PIK mutant unable to bind to the p110alpha catalytic subunit of PI 3-kinase. This mutant had a dominant-negative action on insulin-stimulated glucose transport, since insulin's effect on Glut 4 myc translocation was inhibited in adipocytes expressing mutant p55PIK. Importantly, this dominant-negative mutant was more efficient than wild type p55PIK in associating to IGF-IR and insulin receptor substrate-1 in 293 cells. Taken together, our results show that p55PIK interacts with key elements in the IGF-I signaling pathway, and that these interactions are negatively modulated by PI 3-kinase itself, providing circuitry for regulatory feedback control. PMID- 9415397 TI - Insulin-like growth factor I regulates gonadotropin responsiveness in the murine ovary. AB - The present study shows that insulin-like growth factor I (IGF-I) and FSH receptor (FSHR) mRNAs are selectively coexpressed in a subset of healthy appearing follicles in murine ovaries, irrespective of cycle stage. Aromatase gene expression, a prime marker for FSH effect, is found only in IGF-I/FSHR positive follicles, showing that these are healthy, gonadotropin-responsive follicles. Given the striking coexpression of FSHR and IGF-I, we hypothesized that FSH was responsible for follicular IGF-I expression. We found, however, that granulosa cell IGF-I mRNA levels are not reduced in hypophysectomized (+/-PMSG) or FSH knockout mice, indicating that FSH does not have a major role in regulation of granulosa cell IGF-I gene expression. To test the alternative hypothesis that IGF-I regulates FSHR gene expression, we studied ovaries from IGF I knockout mice. FSHR mRNA was significantly reduced in ovaries from homozygous IGF-I knockout compared with wild type mice and was restored to control values by exogenous IGF-I treatment. The functional significance of the reduced FSHR gene expression in IGF-I knockout ovaries is suggested by reduced aromatase expression and by the failure of their follicles to develop normally beyond the early antral stage. In fact, IGF-I knockout and FSH knockout ovaries appear very similar in terms of arrested follicular development. In summary, we have shown that IGF-I and FSHR are selectively coexpressed in healthy, growing murine follicles and that FSH does not affect IGF-I expression but that IGF-I augments granulosa cell FSHR expression. These data suggest that ovarian IGF-I expression serves to enhance granulosa cell FSH responsiveness by augmenting FSHR expression. PMID- 9415398 TI - Follicle stimulating hormone-regulated expression of serum/glucocorticoid inducible kinase in rat ovarian granulosa cells: a functional role for the Sp1 family in promoter activity. AB - Recently, a family of novel, serine/threonine protein kinases has been identified. One of these transcriptionally inducible, immediate-early genes encodes serum/glucocorticoid inducible-protein kinase, sgk. By in situ hybridization, we show that sgk expression in the rat ovary is selectively localized to granulosa cells. In culture, FSH or forskolin, activators of the protein kinase A (PKA) pathway, rapidly (2 h) and transiently increased sgk mRNA levels in undifferentiated granulosa cells. Sgk mRNA exhibited a biphasic expression pattern, with maximal levels observed at 48 h of FSH/forskolin as granulosa cells differentiate to the preovulatory phenotype. Deletion analyses using sgk promoter-reporter constructs (-4.0 kb to -35 bp) identified a region between -63 and -43 bp that mediated FSH and forskolin-responsive transcription in undifferentiated and differentiated granulosa cells. This G/C-rich region 1) conferred both basal and inducible transcription to the minimal -35 sgk promoter chloramphenicol acetyltransferase reporter construct, 2) specifically bound Sp1 and Sp3 present in granulosa cell extracts, and 3) bound recombinant Sp1. Mutation of 2 bp in this region not only prevented Sp1 and Sp3 binding, but also abolished the PKA-mediated transactivation observed when using the wild type construct. Sp1 and Sp3 DNA-binding activity and protein levels did not change significantly during sgk induction. Collectively, these data indicate that Sp1/Sp3 transactivation of the sgk promoter likely involves regulated, phosphorylation-dependent interaction with other factors. Thus the novel, biphasic induction of sgk that correlates with granulosa cell progression from proliferation to differentiation appears to involve sequential, coordinated actions of FSH, PKA, and transcription factors, including Sp1 and Sp3. PMID- 9415399 TI - A transcriptional silencing domain in DAX-1 whose mutation causes adrenal hypoplasia congenita. AB - The DAX-1 gene encodes an unusual member of the nuclear hormone receptor superfamily. Mutations in the human DAX-1 gene cause X-linked adrenal hypoplasia congenita associated with hypogonadotropic hypogonadism. We have shown that DAX-1 binds to hairpin secondary structures and blocks steroidogenesis in adrenal cells via transcriptional repression of the steroidogenic acute regulatory protein (StAR) promoter. Here we have investigated the molecular mechanism of DAX-1 mediated repression. We show that the DAX-1 C terminus contains a potent transcriptional silencing activity, which can be transferred to a heterologous DNA-binding domain. Deletion analysis and modeling of DAX-1 structure identify two cooperating domains required for the silencing function, one located within helix H3 and the other within H12. The silencing function is cell- and promoter specific. Strikingly, two point mutations (R267P and deltaV269) found in adrenal hypoplasia patients impair silencing. These findings suggest that transcriptional silencing by DAX-1 plays a critical role in the pathogenesis of adrenal hypoplasia congenita. PMID- 9415400 TI - Cloning of human 25-hydroxyvitamin D-1 alpha-hydroxylase and mutations causing vitamin D-dependent rickets type 1. AB - The secosteroid hormone, 1,25-dihydroxyvitamin D [1,25(OH)2D], plays a crucial role in normal bone growth, calcium metabolism, and tissue differentiation. The key step in the biosynthesis of 1,25(OH)2D is its 1 alpha-hydroxylation from 25 hydroxyvitamin D (25-OHD) in the kidney. Because its expression in the kidney is very low, we cloned and sequenced cDNA for 25-OHD-1 alpha-hydroxylase (P450c1 alpha) from human keratinocytes, in which 1 alpha-hydroxylase activity and mRNA expression can be induced to be much greater. P450c1 alpha mRNA was expressed at much lower levels in human kidney, brain, and testis. Mammalian cells transfected with the cloned P450c1 alpha cDNA exhibit robust 1 alpha-hydroxylase activity. The identity of the 1,25(OH)2D3 product synthesized in transfected cells was confirmed by HPLC and gas chromatography-mass spectrometry. The gene encoding P450c1 alpha was localized to chromosome 12, where the 1 alpha-hydroxylase deficiency syndrome, vitamin D-dependent rickets type 1 (VDDR-1), has been localized. Primary cultures of human adult and neonatal keratinocytes exhibit abundant 1 alpha-hydroxylase activity, whereas those from a patient with VDDR-1 lacked detectable activity. Keratinocyte P450c1 alpha cDNA from the patient with VDDR-1 contained deletion/frameshift mutations either at codon 211 or at codon 231, indicating that the patient was a compound heterozygote for two null mutations. These findings establish the molecular genetic basis of VDDR-1, establish a novel means for its study in keratinocytes, and provide the sequence of the key enzyme in the biological activation of vitamin D. PMID- 9415401 TI - Expression and characterization of recombinant type 2 3 alpha-hydroxysteroid dehydrogenase (HSD) from human prostate: demonstration of bifunctional 3 alpha/17 beta-HSD activity and cellular distribution. AB - In androgen target tissues, 3alpha-hydroxysteroid dehydrogenase (3alpha-HSD) may regulate occupancy of the androgen receptor (AR) by catalyzing the interconversion of 5alpha-dihydrotestosterone (5alpha-DHT) (a potent androgen) and 3alpha-androstanediol (a weak androgen). In this study, a 3alpha-HSD cDNA (1170 bp) was isolated from a human prostate cDNA library. The human prostatic 3alpha-HSD cDNA encodes a 323-amino acid protein with 69.9%, 84.1%, 99.4%, and 87.9% sequence identity to rat liver 3alpha-HSD and human type 1, type 2, and type 3 3alpha-HSDs, respectively, and is a member of the aldo-keto reductase superfamily. The close homology with human type 2 3alpha-HSD suggests that it is either identical to this enzyme or a structural allele. Surprisingly, when the recombinant protein was expressed and purified from Escherichia coli, the enzyme did not oxidize androsterone when measured spectrophotometrically, an activity previously assigned to recombinant type 2 3alpha-HSD using this assay. Complete kinetic characterization of the purified protein using spectrophotometric, fluorometric, and radiometric assays showed that the catalytic efficiency favored 3alpha-androstanediol oxidation over 5alpha-DHT reduction. Using [14C]-5alpha-DHT as substrate, TLC analysis confirmed that the reaction product was [14C]-3alpha androstanediol. However, in the reverse reaction, [3H]-3alpha-androstanediol was oxidized first to [3H]-androsterone and then to [3H]-androstanedione, revealing that the expressed protein possessed both 3alpha- and 17beta-HSD activities. The 17beta-HSD activity accounted for the higher catalytic efficiency observed with 3alpha-androstanediol. These findings indicate that, in the prostate, type 2 3alpha-HSD does not interconvert 5alpha-DHT and 3alpha-androstanediol but inactivates 5alpha-DHT through its 3-ketosteroid reductase activity. Levels of 3alpha-HSD mRNA were measured in primary cultures of human prostatic cells and were higher in epithelial cells than stromal cells. In addition, elevated levels of 3alpha-HSD mRNA were observed in epithelial cells derived from benign prostatic hyperplasia and prostate carcinoma tissues. Expression of 3alpha-HSD was not prostate specific, since high levels of mRNA were also found in liver, small intestine, colon, lung, and kidney. This study is the first complete characterization of recombinant type 2 3alpha-HSD demonstrating dual activity and cellular distribution in the human prostate. PMID- 9415402 TI - Estrogen-dependent transcriptional activation and vitellogenin gene memory. AB - The concept of hepatic memory suggests that a gene responds more rapidly to a second exposure of an inducer than it does during the initial activation. To determine how soon estrogen-dependent DNA/protein interactions occur during the primary response, in vivo dimethylsulfate footprinting was carried out using genomic DNA amplified by ligation-mediated PCR. When estrogen was added to disrupted cells from a hormone-naive liver, changes within and around the estrogen response elements occurred within seconds, indicating a direct and rapid effect on this estrogen-responsive promoter that had never before been activated. Because this effect was so rapid relative to the delayed onset of mRNA accumulation during the primary response, run-on transcription assays were used to determine the transcription profiles for four of the yolk protein genes during the primary and secondary responses to estrogen. As with the accumulation of mRNA, the onset of transcription was delayed for all of these genes after a primary exposure to estrogen. Interestingly, after the secondary exposure to estrogen, the vitellogenin I, vitellogenin II, and very low density apolipoprotein II genes displayed a more rapid onset of transcription, whereas the primary and secondary profiles of apolipoprotein B transcription in response to estrogen were identical. Because the apoB gene is constitutively expressed in the absence of estrogen, and the vitellogenins are quiescent before the administration of the hormone, hepatic memory most likely represents a relatively stable event in the transition to an active state of a gene that is committed for tissue-specific expression. PMID- 9415403 TI - Effects of multiple estrogen responsive elements, their spacing, and location on estrogen response of reporter genes. AB - Most highly estrogen-responsive genes possess multiple estrogen-responsive elements (EREs) that act synergistically to activate expression. Synergism between EREs appears to depend on structural features of the EREs and the promoter. To examine the activation process, we cloned single or multiple tandem copies of the consensus ERE into reporter plasmids. These plasmids contained either a chloramphenicol acetyl transferase reporter gene driven by a minimal promoter or a luciferase reporter gene driven by the Simian virus 40 (SV40) promoter. Using MCF-7 human breast cancer cells, we demonstrate that synergism among EREs depends on the number of EREs, their spacing, and the distance of the EREs from the promoter. The induction capacity of EREs falls off slowly with distance from the promoter. Remarkably, multiple EREs can induce effectively and synergize even when they are located more than 2000 nucleotides from the promoter. For EREs located immediately upstream of the promoter, both the distance separating the EREs and the distance to the promoter have to be optimal for synergy. Altering either distance changes the response from synergistic to additive. For distant EREs, presumed to interact by a looping mechanism at the promoter, the length of DNA between the EREs and the promoter is not critical. Synergy among closely spaced EREs that are far from the promoter only requires an optimal distance separating the ERE centers of symmetry. Interestingly, very widely separated EREs can also synergize, presumably also because of their ability to interact by looping. The estrogen response from single or multiple tandem copies of ERE half-palindromes near the SV40 promoter was also tested. The negligible induction capacity of a single half-site was not significantly increased in multiple sites. The biological role of half-EREs is not apparent in the system employed here. PMID- 9415404 TI - Loss of an estrogen receptor isoform (ER alpha delta 3) in breast cancer and the consequences of its reexpression: interference with estrogen-stimulated properties of malignant transformation. AB - Comparison of mRNA ratios of a non-DNA-binding estrogen receptor (ER(alpha)) isoform, missing exon 3 (ER(alpha)delta3), to the full-length ER(alpha), in normal breast epithelium to that in primary breast cancers and breast cancer cell lines revealed a 30-fold reduction of this ratio in cancer cells (P < 0.0001). To test what functions may have been affected by the loss of ER(alpha)delta3, stable clones of MCF-7 cells expressing ectopic ER(alpha)delta3 protein, at the range of physiological ER(alpha), were generated. In vector-transfected controls the ER(alpha)delta3-mRNA and protein were less than 10% while in the ER(alpha)delta3 expressing clones, ER(alpha)delta3-mRNA and protein ranged from 36-76% of the total ER(alpha). Estrogen (E2) stimulated the expression of pS2-mRNA in pMV7 vector control cells, but the stimulation was reduced by up to 93% in ER(alpha)delta3-expressing clones. In addition, several properties associated with the transformed phenotype were also strongly affected when ER(alpha)delta3 protein was reexpressed. Compared with vector-transfected control cells, the saturation density of the ER(alpha)delta3-expressing clones was reduced by 50 68%, while their exponential growth rate was only slightly (14.5 +/- 5%) lower. The in vivo invasiveness of the ER(alpha)delta3-expressing cells was significantly reduced (P = 0.007) by up to 79%. E2 stimulated anchorage independent growth of the pMV7 vector control cells, but reduced it to below baseline levels in ER(alpha)delta3 clones. The reduction of the pS2 response to E2 in the ER(alpha)delta3-expressing clones and the E2 block of anchorage independent growth to below baseline were more pronounced than expected from the dominant negative function of ER(alpha)delta3. These observations suggest that E2 may activate an additional ER(alpha)delta3-dependent inhibitory pathway. The drastic reduction of ER(alpha)delta3 to ER(alpha) ratio in breast cancer, and the fact that when present in breast cancer cells this isoform leads to a suppression, rather than enhancement, of the transformed phenotype by E2 suggests that the regulation of ER(alpha)-mRNA splicing may need to be altered for the breast carcinogenesis to proceed. PMID- 9415405 TI - Stress-induced transcriptional regulation in the developing rat brain involves increased cyclic adenosine 3',5'-monophosphate-regulatory element binding activity. AB - The cAMP-regulatory element (CRE) binding protein (CREB) functions as a trans acting regulator of genes containing the CRE sequence in their promoter. These include a number of critical genes, such as CRF, involved in the hypothalamic response to stressful stimuli in the adult. The ability of the developing rat (during the first 2 postnatal weeks) to mount the full complement of this stress response has been questioned. We have previously demonstrated the stress-induced up-regulation of the transcription of hypothalamic CRF during the second postnatal week in the rat. The focus of the current study was to explore the mechanism of transcriptional regulation in response to stress through the physiological induction of transcriptional trans-activators that bind to the CRE in the developing rat brain. CRE-binding activity was detected via gel shift analysis in extracts from both the hypothalamus and the cerebral cortex of the developing rat. CREB was identified in these extracts by Western blot analysis and was shown to be the major contributor to the CRE-binding activity by gel shift analysis with two specific antibodies directed against CREB. After acute hypothermic stress, the abundance of CRE-binding activity (but not of total immunoreactive CREB), increased in hypothalamic extracts. This enhanced CRE binding activity was blocked by an antiserum directed against CREB and was accompanied by an apparent increase in CREB phosphorylation. These results indicate that posttranslational enhancement of CRE-binding activity is likely to constitute an important mechanism for up-regulation of genes possessing the CRE sequence in the developing rat hypothalamus by adverse external signals. PMID- 9415406 TI - Characterization of receptor interaction and transcriptional repression by the corepressor SMRT. AB - SMRT (silencing mediator of retinoic acid and thyroid hormone receptor) and N-CoR (nuclear receptor corepressor) are two related transcriptional corepressors that contain separable domains capable of interacting with unliganded nuclear receptors and repressing basal transcription. To decipher the mechanisms of receptor interaction and transcriptional repression by SMRT/N-CoR, we have characterized protein-protein interacting surfaces between SMRT and nuclear receptors and defined transcriptional repression domains of both SMRT and N-CoR. Deletional analysis reveals two individual nuclear receptor domains necessary for stable association with SMRT and a C-terminal helix essential for corepressor dissociation. Coordinately, two SMRT domains are found to interact independently with the receptors. Functional analysis reveals that SMRT contains two distinct repression domains, and the corresponding regions in N-CoR also repress basal transcription. Both repression domains in SMRT and N-CoR interact weakly with mSin3A, which in turn associates with a histone deacetylase HDAC1 in a mammalian two-hybrid assay. Far-Western analysis demonstrates a direct protein-protein interaction between two N-CoR repression domains with mSin3A. Finally we demonstrate that overexpression of full-length SMRT further represses basal transcription from natural promoters. Together, these results support a role of SMRT/N-CoR in corepression through the utilization of multiple mechanisms for receptor interactions and transcriptional repression. PMID- 9415407 TI - Extracellular signal-regulated kinase-1 and -2 respond differently to mitogenic and differentiative signaling pathways in myoblasts. AB - In this report we show that extracellular signal-regulated kinase-1 and -2 (ERK-1 and -2) respond differently to signals that elicit proliferation and/or differentiation of myoblasts using the C2C12 cell line and nondifferentiating mutant NFB4 cells derived from them. Induction of differentiation by withdrawal of serum rendered ERKs in C2C12 myoblasts relatively insensitive to restimulation by serum. Instead, myogenic differentiation of C2C12 cells was associated with sustained activation of ERK-2 dependent on the insulin-like growth factor II (IGF II) autocrine loop. By contrast, mutant NFB4 cells cultured under the same conditions remained proliferative and demonstrated robust activation of ERKs in response to serum. Similarly, a Gi-dependent signaling pathway induced activation of ERKs in NFB4 cells, but not in C2C12 cells, after stimulation by lysophosphatidic acid (LPA). In NFB4 cells partially rescued by prolonged IGF-I treatment, ERK activity remained responsive to Gi-dependent LPA stimulation, whereas rescue of NFB4 cells by constitutive expression of myogenin or MyoD, associated with activation of the IGF-II autocrine loop, rendered the Gi signaling pathway refractory to LPA stimulation. Relatively high levels of G(alpha i2) were detected in NFB4 cells and IGF-I treated NFB4 cells, which correlated with responsive Gi signaling. Activation of the IGF-II autocrine loop in C2C12 and NFB4 myoblasts or treatment with IGF-II was associated with loss of G(alpha i2) and inhibition of Gi-dependent signaling. Thus, IGF-I and IGF-II activate distinct signaling cascades, with IGF-II eliciting a stronger differentiation effect correlated with down-regulation of G(alpha i2) protein. Short-term stimulation of NFB4 cells with IGF-I, a mitogenic signal for myoblasts, also induced ERK-1 and -2 activation. Transient stimulation of NFB4 cells with IGF-I while blocking activation of Gi-proteins is with pertussis toxin resulted in preferential activation of ERK-2 characteristic of differentiated C2C12 cells, suggesting that proliferation induced by IGF-I is Gi-dependent and separable from the IGF-I-signaling pathway that leads to differentiation. PMID- 9415408 TI - Localization of agonist- and antagonist-binding domains of human corticotropin releasing factor receptors. AB - The CRF receptors, CRFR1 and CRFR2, are members of the G protein-coupled receptor superfamily. Despite their considerable sequence similarity, CRFR1 and CRFR2 have quite different affinities for the peptide ligand rat/human CRF. Previous studies using chimeric receptors between human CRFR1 and CRFR2 have identified three potentially important regions in the second and third extracellular domains of CRF receptor for the binding of rat/human CRF. The present report further demonstrates that these same three regions also affect the binding of urocortin and sauvagine, two other members of the CRF peptide family, albeit to different extents. We also show that a fourth region in the third extracellular domain, Asp254, has been identified to be important for sauvagine but not CRF or urocortin binding. Thus, the three peptide ligands not only interact with a different set of regions on CRFR1 and CRFR2 but also differentially interact with some of the same regions. These data could, at least in part, account for the much higher affinity of CRFR2 for urocortin and sauvagine compared with rat/human CRF. We have also identified two amino acid residues, His199 in the third transmembrane domain and Met276 in the fifth transmembrane domain, that are important for binding the non-peptide high-affinity CRFR1 antagonist NBI 27914. Mutations of His199 and Met276 to the corresponding amino acids in CRFR2 each decreased the binding affinity of NBI 27914 for CRFR1 by 40- and 200-fold, respectively. This suggests that the transmembrane regions are critically important in forming the binding pocket for the nonpeptide antagonist. PMID- 9415409 TI - Drug retention, efflux, and resistance in tumor cells. AB - Multiple drug resistance (MDR) in tumor cells has been related to the expression of transport proteins which alter cellular drug transport and distribution. Three different genes (mdr, MRP, and LRP) and their products have been implicated in MDR. Several fluorescent dyes have been used to monitor the effect of these transport proteins on drug retention, as well as for screening of drugs which block drug efflux and thus enhance cellular drug retention and cytotoxicity. The present review summarizes current knowledge about MDR phenotypic markers and techniques available for study of MDR by flow cytometry. PMID- 9415410 TI - Use of a membrane-localized green fluorescent protein allows simultaneous identification of transfected cells and cell cycle analysis by flow cytometry. AB - The simultaneous detection of the green fluorescent protein (GFP) and DNA content using propidium iodide (PI) by flow cytometry is made difficult because of the unique nature of these 2 fluorogenic reagents. For PI to enter cells efficiently and to stain DNA quantitatively, the cells must first be permeabilized; ethanol treatment is a routine method to achieve this. However, this permeabilization step causes GFP, which is normally found in the cytoplasm, to leach out of the cells. Although the use of paraformaldehyde-based fixatives allows GFP to be maintained in cells and retain its fluorescence even after ethanol permeabilization, the protocol we commonly employ results in inefficient PI staining and poor quality DNA histograms. To circumvent these difficulties, we have employed a GFP-fusion protein which localizes to the cellular membrane and as such is retained in cells upon ethanol permeabilization without prior fixation. This allows the GFP signal to be detected in cells treated with ethanol in preparation for PI staining and cell cycle analysis. This property facilitates the use of GFP as a marker for transfected cells in experiments designed to characterize the effects of ectopic expression of cellular or viral genes on cell cycle progression. PMID- 9415411 TI - Solid phase cytometry allows rapid in situ quantification of human papilloma virus infection in biopsy material. AB - Solid phase cytometry would be an asset for many histological and cytological studies. Current microscope-based cytometers and image analysis systems are too slow to analyze specimens several millimeters wide. We have recently shown that a rapid wide area laser scanning device that operates on solid supports has a linear response. We assess it here for solid phase cytometry. Each cell detected by the cytometer can be automatically positioned for visual observation in the field of an epifluorescence microscope (conventional or confocal) in which the stage is driven by the instrument's computer. We were able to detect and map human papillomavirus-infected cells labeled by fluorescent in situ hybridization in cervical condyloma biopsies. We could quantify the fluorescence emitted by these cells and show differences of up to 35-fold in fluorescence intensity between individual cells. These differences in intensity might reflect differences in viral copy number. The potential of the system to provide fast, reliable and reproducible analyses of solid tissue samples is discussed. PMID- 9415412 TI - Assessment of E. coli and Salmonella viability and starvation by confocal laser microscopy and flow cytometry using rhodamine 123, DiBAC4(3), propidium iodide, and CTC. AB - Assessment of cell viability using methods which do not require cell culture is essential in the field of aquatic microbiology, since many bacteria known to be present in aquatic environments cannot be grown in culture. The study of bacterial biofilms, which previously needed an epifluorescent microscope, has recently been enhanced by the use of flow cytometry and confocal scanning laser microscopy (CSLM). A method based on the combination of several membrane potential related dyes, a membrane integrity dye and a redox probe was used to measure cell viability by flow cytometry and confocal laser microscopy. Rhodamine propidium iodide (PI) double staining was used to discriminate viable from nonviable cells in CSLM observations. Membrane depolarization during E. coli and Salmonella starvation measured by DiBAC4(3) incorporation (flow cytometry and CSLM) was found to be in concordance with respiratory activity as detected by a tetrazolium salt (CTC) reduction. PMID- 9415414 TI - In situ detection of apoptosis during embryogenesis with annexin V: from whole mount to ultrastructure. AB - Apoptosis is of paramount importance during embryonic development. This insight stems from early studies which correlated cell death to normal developmental processes and now has been confirmed by linking aberrant cell death patterns to aberrant development. Linking apoptosis to the phenotype of a developing organism requires spatial information on the localization of the dying cells, making in situ detection essential This prerequisite limits the tools available for such studies (1) to vital dyes, which can be detected at the whole mount level only; (2) to detection based upon apoptotic morphology by routine light microscopy and electron microscopy; and (3) to staining for apoptosis associated DNA fragmentation via, e.g., the TUNEL procedure, which marks cells in a relative late phase of apoptosis. New apoptosis markers need to be specific and should preferably detect cells early during this process. In the present study we show that the recently discovered in vitro marker of apoptosis, Annexin V meets these requirements for in vivo detection. Through intracardiac injections of biotin labeled Annexin V, a Ca2+ dependent phosphatidylserine binding protein, we were able to visualize apoptotic cells derived from each germ layer in the developing mouse embryo from the whole mount level up to the ultrastructural level. Double labeling on paraffin sections for both this method and TUNEL revealed that cells become Annexin V-biotin labeled early during the process of apoptosis. PMID- 9415413 TI - Monitoring early cellular responses in apoptosis is aided by the mitochondrial membrane protein-specific monoclonal antibody APO2.7. AB - A recently described mitochondrial membrane protein-specific monoclonal antibody, APO2.7, was examined for monitoring early apoptotic responses in anti-CD95 (7C11) induced Jurkat cells. Jurkat cells were harvested at 1.5, 3, 4.5, 6, 12, and 18 h after induction of apoptosis, and APO2.7 antibody monitored in unprocessed (no permeabilization agent used prior to staining) and processed (permeabilized prior to staining) cells. Light-scatter changes (decreased forward-scatter and increased side-scatter) by flow cytometry were observed after 3 h, and detection of cell permeability in unprocessed cells, as measured by light microscopic examination of Trypan blue-stained cells and flow cytometric detection of tubulin, showed little change until after 6 h. In addition, unprocessed cells stained with APO2.7 antibody showed little increase in staining until after 6 h following induction of apoptosis, when DNA fragmentation was demonstrated by flow cytometry and gel electrophoresis; however, processed cells stained with APO2.7 antibody showed significant increase in staining after 1.5 h. Detection, using annexin V and flow cytometry, of phospholipid membrane asymmetry from exposure of phosphatidylserine showed greater, apparent nonspecific staining in noninduced cells as compared to the other markers of apoptosis, but nearly paralleled the results of APO2.7 staining in processed cells from 3-18 h following CD95 induction of apoptosis. The data presented herein indicate that the mitochondrial membrane protein-specific antibody, APO2.7, is useful as a marker for the detection of apoptotic cells. PMID- 9415415 TI - Activated cell cycle checkpoints in epirubicin-treated breast cancer cells studied by BrdUrd-flow cytometry. AB - Genetic alterations, such as p53 mutations, may affect a tumour's response to chemotherapy. We have treated two human breast cancer cell lines that differ in p53 status with epirubicin in order to study if there are differences in cell cycle kinetic response. MCF-7 cells express wild-type p53, while SK-BR-3 cells express only a mutated form of p53. The transition of cells from one cell cycle stage to another was studied by a bromodeoxyuridine (BrdUrd)-flow cytometry (FCM) method. MCF-7 cells showed a block in the G1 phase after treatment with 50 nM epirubicin for 24 hours, in agreement with the actions of p53 at the G1 checkpoint. SK-BR-3 cells, on the other hand, progressed through the G1 checkpoint and were blocked in late S and G2 phases, presumably due to the activation of a later checkpoint. In addition, studies of the mRNA levels of p53 and its effector gene p21 revealed that although both cell lines expressed p53 mRNA, a marked difference in the mRNA levels of p21 was seen. A dramatic increase in the level of p21 mRNA was seen in epirubicin-treated MCF-7 cells, while no such increase was seen in SK-BR-3 cells. PMID- 9415416 TI - 8 color, 10-parameter flow cytometry to elucidate complex leukocyte heterogeneity. AB - We developed the chemistry, instrumentation, and software technologies needed to measure, simultaneously and independently, eight different fluorescent molecules on individual cells. Conjugation of these fluorochromes to monoclonal antibodies is straightforward; all immunofluorescence staining is accomplished with direct stains only. We built a hybrid flow cytometer with eight fluorescence detectors and two light scatter channels, with excitation provided by three spatially separated laser beams emitting at 407 nm, 488 nm, and 595 nm. The fluorescence compensation required to make the data orthogonal is of sufficient complexity that it cannot be performed manually; thus, we use software to compensate the data post hoc, based on data collected from singly stained compensation control samples. In this report, we evaluate the 8 color staining technology. Of the seven fluorochromes other than fluorescein, six have a useful brightness at least as great as fluorescein. Three of the fluorochromes (phycoerythrin, allophycocyanin, and the Cy5 resonance energy tandem of phycoerythrin) are considerably brighter than fluorescein and are useful for detecting antigens expressed at low levels. Finally, we show the power and utility of the 8 color, 10-parameter technology using staining experiments on both human and murine immune systems. PMID- 9415417 TI - Lymphocyte subset analysis on frozen whole blood. AB - An approach to perform lymphocyte subset analysis on frozen-thawed whole blood (F/T WB) is described. WB from 24 human immunodeficiency virus type 1 (HIV-1) seropositive individuals and 21 controls was analyzed fresh and after frozen storage (with or without dimethyl sulfoxide) at -80 degrees C, in liquid nitrogen (LN2), and at -20 degrees C. Analysis of F/T WB utilized 3-color flow cytometry with CD45 and right angle light scatter gating. Absolute cell counts were obtained for 30 samples by using staining tubes containing internal bead standards [TruCount, Becton Dickinson Immunocytometry Systems (BDIS), San Jose, CA]. The mean difference between CD3+4+ percentages for F/T (-80 degrees C storage for up to 1 year) and fresh WB was less than -0.2% (95% limits +/-3%, P = 0.5) with 39 of 45 (87%) results falling within 2% of the fresh values (P = 0.74). Absolute CD3+4+ cell counts for F/T WB were generally lower than corresponding results for fresh aliquots (median difference was 33 cells/microl, P < 0.0001), but the results were highly correlated (r2 = 0.975, P < 0.0001). Results were more variable, although still highly correlated, for CD3+8+ cells, and with other freezing and storage conditions. It is concluded that lymphocyte subset analysis using F/T WB yields comparable results to fresh samples, which should prove useful for a number of practical applications. PMID- 9415418 TI - Lymphocyte subsets and expression of differentiation markers in blood and lymphoid organs of rhesus monkeys. AB - Rhesus macaques are invaluable experimental animals in biomedical research. Using three color flow cytometry, we screened anti-human antibodies for crossreactivity with macaque cells in order to determine the distribution of functionally important lymphocyte subsets in blood, lymph nodes (LN), and spleen. NK-cells are almost completely absent in LN. The percentage of B-cells expressing CD80, CD86, and the level of expression of CD20 is higher in blood than in LN. In contrast, a higher proportion of B-cells in LN stains positive for CD21 and CD35. Whereas the number of CD29hi expressing T-cells is lower, CD69 is expressed on more T-cells in LN than in blood. About one-third of CD8+ T-cells in blood are CD28-, a subset with a unique pattern of antigen expression which cannot be found in LN. In contrast to humans, a relatively high proportion of T-cells in blood also express the co-stimulatory molecules CD80 and CD86. With increasing age, the proportion of B-cells in blood declines, whereas the percentage of T-cells rises. In addition, the proportion of CD29hi expressing T-cells increases among both the CD4+ and CD8+ subsets. PMID- 9415419 TI - Radially symmetric excitation and collection optics for flow cytometric sorting of aspherical cells. AB - We report on a new optical configuration for sorting flow cytometers which is optimized for the illumination and collection of light from aspherical cells. This design provides radially symmetric illumination and detection of asymmetric particles while retaining the sort capability of a jet-in-air (or cylindrical cuvette) design. A paraboloidal reflector, symmetrical about the sample stream, both focuses a laser excitation beam and collects cell scatter and fluorescence from the inspection point. The performance of the new optical configuration has been tested and compared to that of a modified commercial flow cytometer, which uses a forward-side fluorescence collection geometry. For fluorescence measurements on calibration microspheres the new system produces histograms with similar coefficients of variation to those obtained with the modified conventional cytometer. Optical artifacts apparent in measurements on flat cells, such as blood cells and mammalian sperm, using conventional optics, are eliminated by the new configuration. Analysis of chinchilla sperm yields a dual peaked histogram population that has a coefficient of variation and X-Y split which matches that for gated (oriented) fraction of the sample as measured by the conventional system. Bovine sperm, which are larger and flatter than chinchilla sperm, also produce a single population which, when low sample-to-sheath differential pressures are used, has coefficients of variation matching those for an oriented subpopulation as measured by conventional optics. This new optical configuration presents an alternative technique for measuring aspherical cells independent of their orientation, with the potential for higher analysis rates and improved efficiency compared to other optical systems. PMID- 9415420 TI - 1-NGFR receptor is a new flow cytometric tool for rapid cell cycle-correlated gene therapy complementation studies in viable cells. AB - Control of successful genetic complementation of cellular defects in heterogenous cell populations requires biochemical selection markers or cell analytical specifiers. With available gene therapy technologies, only a fraction of a cell population is transfected or transduced. We applied and optimized a novel dual laser flow cytometric technique to analyze immediately the genetic complementation of cells dysregulated in cell proliferation and DNA repair. A novel bicistronic retrovirus carrying the normal Fanconi anemia gene (group C) and the cell surface marker gene l-NGFR was constructed to analyze the normalization of the G2-phase cell cycle defect in DNA-repair-deficient FA(C) lymphoblastoid cells after transduction. Using a dual-laser multiparameter technology, we 1) analyzed the cell-cycle distribution of viable/dead cells using Hoechst 33342 (with ultraviolet light), 2) identified the genetically complemented cells by FITC antibody labeling of the novel l-NGFR surface marker (488 nm), and 3) recorded mitomycin C-induced cell death in nontransduced and transduced cells by propidium iodide. Artificial l-NGFR expression is high and similar in ontogenetically and phylogenetically different cell populations. With this novel l-NGFR marker technology, the success of gene therapy of cell-cycle dysregulation in even small subpopulations of cells can be recorded within 48 h. In addition to improved cell analysis, l-NGFR surface marker expression can also be used for rapid generation of pure cell populations by column cell separation technologies. PMID- 9415421 TI - Nuclear dyes and cytoplasmic staining. PMID- 9415422 TI - Keratin 8 and 18 expression in mesenchymal progenitor cells of regenerating limbs is associated with cell proliferation and differentiation. AB - Keratins are considered markers of epithelial differentiation. In lower vertebrates, however, immunoreactivity for keratin 8 and 18 has been reported in nonepithelial cells, particularly in mesenchymal progenitor cells of regenerating complex body structures. To confirm that such reactivity does indeed reflect keratin expression and to investigate their possible role in regeneration, we have isolated clones coding for the newt homologues of keratin 8 and 18 (NvK8 and NvK18, respectively) and studied their distribution and changes in their expression following experimental manipulations. Analysis of NvK8 and NvK18 transcripts confirms that K8 and K18 are expressed in the blastemal cells of regenerating newt limbs and that their expression is first observed 3-5 days after amputation, when the blastemal cells start to proliferate under the influence of the nerve, whose presence is essential for regeneration to proceed. In contrast, no induction of these keratins is observed following amputation of a larval limb at a stage when organogenesis is proceeding in a nerve-independent manner. To establish whether there is a causal relationship between keratin expression and cell proliferation in the adult limb blastema, we have investigated whether their expression is nerve-dependent and whether suppression of their expression in cultured blastemal cells affects cell division and differentiation. Analysis of keratins in denervated limbs demonstrates that the nerve is not necessary to induce their expression. However, treatment of cultured blastemal cells with K8 and K18 anti-sense oligonucleotides significantly decreases DNA synthesis and induces changes in cell morphology, suggesting that expression of these keratins during regeneration may be necessary for the maintenance of the undifferentiated and proliferative state of blastemal cells. PMID- 9415423 TI - Establishment of stem cell identity in the Drosophila germline. AB - In the adult Drosophila ovary the continuous production of eggs depends upon a small group of stem cells located at the anterior tip of the germarium. These stem cells divide asymmetrically to self renew and to generate a cystoblast, which in females is committed to the oocyte differentiation pathway. While much is known about the development of poststem cell cystoblasts, little is known about when stem cells are formed or how their identity is initially established. To investigate these questions we have used the P-M hybrid dysgenesis syndrome as a tool for ablating the "pre-stem cell" progenitors of the stem cells. Our experiments indicate that the pre-stem cells in females assume stem cell identity during the early pupal stage. Our results also suggest a model in which at least two pre-stem cells must be present within an ovariole for the specification of stem cell fate. When only a single pre-stem cell is sequestered by an ovariole, this cell does not assume stem cell identity, but instead follows the cystoblast cystocyte differentiation pathway. PMID- 9415424 TI - Expression patterns of bone morphogenetic proteins (Bmps) in the developing mouse tooth suggest roles in morphogenesis and cell differentiation. AB - Bone morphogenetic proteins (BMP) are secretory signal molecules which have a variety of regulatory functions during morphogenesis and cell differentiation. Teeth are typical examples of vertebrate organs in which development is controlled by sequential and reciprocal signaling between the epithelium and mesenchyme. In addition, tooth development is characterized by formation of mineralized tissues: the bone-like dentin and cementum as well as epithelially derived enamel. We have performed a comparative in situ hybridization analysis of the expression of six different Bmps (Bmp-2 to Bmp-7) starting from initiation of tooth development to completion of crown morphogenesis when dentine and enamel matrices are being deposited. Bmps-2, -4, and -7 were frequently codistributed and showed marked associations with epithelial-mesenchymal interactions. Their expression shifted between the epithelium and mesenchyme starting from the stage of tooth initiation. They were subsequently expressed in the enamel knot, the putative signaling center regulating tooth shape. Their expression domains prior to and during the differentiation of the dentine-forming odontoblasts and enamel forming ameloblasts was in line with functions in regulation of cell differentiation and/or secretory activities of the cells. The expression of Bmp-3 was confined to mesenchymal cells, in particular to the dental follicle cells which give rise to the cementoblasts, forming the hard tissue covering the roots of teeth. Bmp-5 was expressed only in the epithelial ameloblasts. It was upregulated as the cells started to polarize and intense expression continued in the secretory ameloblasts. Bmp-6 was expressed only weakly in the dental mesenchyme during bud and cap stages. Our results are in line with regulatory functions of Bmps at all stages of tooth morphogenesis. Bmps-2, -4, and -7 are conceivably parts of signaling networks regulating tooth initiation and shape development. They as well as Bmp-5 may be involved in the induction and formation of dentine and enamel, and Bmp-3 in the development of cementum. The remarkable overlaps in the expression domains of different Bmp genes may implicate functional redundancy and/or formation of active heterodimers between different BMPs. PMID- 9415425 TI - Epidermal ectoderm is required for full elevation and for convergence during bending of the avian neural plate. AB - Previous studies suggest that bending of the neural plate requires the juxtaposition of neural plate and non-neuroepithelial tissues. The current study examines the role of one of these tissues, the epidermal ectoderm, in bending. Chick blastoderms were harvested from fertile eggs incubated for 24 hr and cultured dorsal-side-up on agar-albumen substrates. In one experiment, a rectangular flap of epidermal ectoderm on one side of each blastoderm was separated from underlying layers and gently reflected onto the area opaca; a fragment of tungsten wire was placed on top of the flap to hold it down and to prevent healing. Embryos were then allowed to develop in a humidified incubator for 2-18 hr. Asymmetric neurulation was observed between the operated and control sides as early as 2 hr after surgery. The amount of asymmetry was quantified in serial transverse sections from embryos collected 8 hr after surgery. Elevation of the lateral edge of the neural plate on the operated side averaged one half to two thirds of that on the control side, and convergence of the operated side around the dorsolateral hinge point toward the dorsal midline did not occur. These results demonstrate that epidermal ectoderm is required for full elevation and for convergence during bending. In another experiment, lateral epidermal ectoderm was removed, leaving only a medial strip consisting of both the epidermal component of the future neural fold and flanking future epidermis. This experiment revealed that although epidermal ectoderm is necessary for full elevation and for convergence of the neural folds, a medial strip of epidermal ectoderm is sufficient to drive bending. Collectively, these results further support the idea that neurulation is a multifactorial process driven by both intrinsic and extrinsic factors acting in concert. PMID- 9415426 TI - Epithelium is required for maintaining FGFR-2 expression levels in facial mesenchyme of the developing chick embryo. AB - In the developing chick embryo, fibroblast growth factor-2 (FGFR-2) expression patterns correlate with outgrowth of facial prominences. Frontonasal mass prominences that form the pre-nasal cartilage and upper beak express high levels of FGFR-2 receptor, whereas maxillary prominences that form the flattened corners of the beak and palatal shelves express low FGFR-2 transcript levels. Facial epithelium is an abundant source of FGFs and is required to support outgrowth of mesenchymal tissue, including cartilage rod formation. Because FGFR-2 is highly expressed in regions of facial outgrowth and because epithelium is required for outgrowth of facial prominences, epithelium could be required to maintain FGFR-2 transcripts in facial mesenchyme. To test this hypothesis, we removed epithelium to inhibit outgrowth of regions of the embryonic face, grafted frontonasal mass and maxillary prominences into a host limb bud, and then examined changes in FGFR 2 expression using in situ hybridization. We also hybridized adjacent sections with collagen II probe to identify regions undergoing chondrogenesis. Our results indicate that removal of epithelium from frontonasal mass led to a decrease in FGFR-2 and collagen II expression 24 hr after grafting to host and that neither FGFR-2 nor collagen II expression increased to expected levels at 48 hr. These results suggest that there are signals in the epithelium required for increasing FGFR-2 and collagen II gene transcription, and the expression of these genes are linked to outgrowth of facial prominences. PMID- 9415427 TI - Changing patterns of gap junctional intercellular communication and connexin distribution in mouse epidermis and hair follicles during embryonic development. AB - In the mouse embryo between embryonic days 12 (E12) and 16, regular arrays of epidermal placodes on the mystacial pad develop into whisker follicles. This system was chosen for analysis of gap junctional intercellular communication during differentiation. The patterns of communication were studied by microinjection of the tracers Lucifer yellow-CH (LY-CH) and neurobiotin (NB), while immunofluorescent staining was used to study distribution of connexins 26 and 43. Extensive communication was seen between keratinocytes in developing hair pegs or, in later-stage hair follicles, in the germinative matrix. Coupling between adjacent hair pegs via interfollicular epidermis was not observed. Coupling also became restricted as follicular cells differentiated to form outer root sheath, inner root sheath, and hair shaft. Extensive gap junctional coupling is characteristic of keratinocytes that are rapidly proliferating (as in hair pegs and germinative matrix). Follicular keratinocytes commence differentiation shortly before restriction of gap junctional coupling becomes evident. Dermal mesenchymal cells undergoing different modes of differentiation also exhibit differences in gap junctional coupling, as evidenced by poor transfer of LY-CH between cells in dermal condensations of hair follicles compared with extensive transfer elsewhere in the dermis. LY-CH and NB were not transferred between epidermal or follicular epithelium and mesenchyme, arguing against a direct role for gap junctions permeable to known second messenger molecules or nucleotides in epithelial-mesenchymal interactions in this system. The distribution of connexins 26 and 43 in epidermis and hair follicles changed during differentiation but there was no correlation with changing patterns of dye transfer, indicating an unexpected degree of complexity in the relationship between gap junctional intercellular communication and connexin protein distribution during development. PMID- 9415429 TI - Differential expression of five laminin alpha (1-5) chains in developing and adult mouse kidney. AB - The nature of the laminin alpha chains in the embryonic and adult kidney is still being debated. The present study attempted to clarify this issue by immunofluorescence study using monoclonal antibodies against mouse alpha1, alpha2, and alpha5 chains and in situ hybridization for the alpha2, alpha3B, alpha4, and alpha5 mRNAs. Novel alpha1 chain-specific monoclonal antibodies against E8 fragment revealed a restricted distribution of alpha1 chain in a subset of epithelial basement membranes in the embryo, in agreement with previous mRNA data. The alpha2 mRNA was produced by mesenchyme, although the protein was deposited in epithelial basement membranes. The alpha3B mRNA was found only in a small subset of endothelial cells. The alpha4 mRNA was found transiently in embryonic mesenchyme, with particularly high levels in condensed mesenchyme, close to the tips of the ureteric tree where tubulogenesis is initiated. The alpha5 mRNA was strongly expressed by ureter epithelium but not expressed at early stages of tubulogenesis. Immunofluorescence verified low levels of the alpha5 chain in the early stages of tubulogenesis. However, during the capillary loop stage, the alpha5 chain became strongly expressed in the developing glomerular basement membrane, which matches the in situ hybridization results. During subsequent maturation of the kidney, the alpha5 chain became ubiquitously expressed in basement membranes. Overall, the alpha5 chain exhibited the broadest pattern of expression, followed by the alpha1 chain, particularly in the adult stage. These chains were the only ones produced by epithelial cells. Although some basement membranes contained several alpha chains, we failed to detect any of the five studied chains in some basement membranes. Thus, the identity of the alpha chains of many embryonic kidney blood vessels and several basement membranes in the inner medulla in the developing and adult kidney remain unclear. PMID- 9415428 TI - Genetic patterning of the developing mouse tail at the time of posterior neuropore closure. AB - Posterior neuropore (PNP) closure coincides with the end of gastrulation, marking the end of primary neurulation and primary body axis formation. Secondary neurulation and axis formation involve differentiation of the tail bud mesenchyme. Genetic control of the primary-secondary transition is not understood. We report a detailed analysis of gene expression in the caudal region of day 10 mouse embryos during primary neuropore closure. Embryos were collected at the 27-32 somite stage, fixed, processed for whole mount in situ hybridisation, and subsequently sectioned for a more detailed analysis. Genes selected for study include those involved in the key events of gastrulation and neurulation at earlier stages and more cranial levels. Patterns of expression within the tail bud, neural plate, recently closed neural tube, notochord, hindgut, mesoderm, and surface ectoderm are illustrated and described. Specifically, we report continuity of expression of the genes Wnt5a, Wnt5b, Evx1, Fgf8, RARgamma, Brachyury, and Hoxb1 from primitive streak and node into subpopulations of the tail bud and caudal axial structures. Within the caudal notochord, developing floorplate, and hindgut, HNF3alpha, HNF3beta, Shh, and Brachyury expression domains correlate directly with known genetic roles and predicted tissue interdependence during induction and differentiation of these structures. The patterns of expression of Wnt5a, Hoxb1, Brachyury, RARgamma, and Evx1, together with observations on proliferation, reveal that the caudal mesoderm is organised at a molecular level into distinct domains delineated by longitudinal and transverse borders before histological differentiation. Expression of Wnt5a in the ventral ectodermal ridge supports previous evidence that this structure is involved in epithelial-mesenchymal interaction. These results provide a foundation for understanding the mechanisms facilitating transition from primary to secondary body axis formation, as well as the factors involved in defective spinal neurulation. PMID- 9415431 TI - Spatial and temporal expression of the beta1D integrin during mouse development. AB - The beta1D protein is a recently characterized isoform of the integrin beta1 subunit that is present in cardiac and skeletal muscles. In this study, we have examined the expression of beta1D in different types of skeletal muscle and in cardiac muscle and studied its distribution during mouse development, using new monoclonal antibodies specific for beta1D. Immunoprecipitation studies revealed that, while beta1A is strongly expressed in proliferating C2C12 myoblasts, beta1D is only expressed after their differentiation to myotubes. In these myotubes, beta1D is associated with different alpha subunits, namely alpha3A, alpha5, alpha7A, or alpha7B. Initially, during embryogenesis, the alpha1A subunit is the only beta1 variant expressed in skeletal and cardiac muscle. The beta1D subunit is first detected in skeletal muscle at E17.5, whereas in cardiac muscle its expression begins around the time of birth. Later the expression of beta1A in skeletal and cardiac muscle becomes restricted to capillary cells, whereas beta1D eventually becomes the only variant expressed in adult cardiac and skeletal muscle cells. The switch from the beta1A to the beta1D subunit in cardiac muscle cells coincides with the expression of alpha7. In adults there is a distinct concentration of beta1D at the myotendinous junctions of muscle fibers and at costameres in both cardiac and skeletal muscle. In addition, beta1D is present at intercalated discs in cardiac muscle and at neuromuscular junctions in skeletal muscle cells. The amount of beta1D in different types of skeletal muscle (fast, slow, and mixed-type) was similar, but cardiac muscle expressed almost five times as much of this protein. We suggest that beta1D plays a role in the maintenance of the cytoarchitecture of mature muscle and in the functional integrity of the muscle cells. PMID- 9415430 TI - Expression of GDNF and its receptors in developing tooth is developmentally regulated and suggests multiple roles in innervation and organogenesis. AB - Glial cell line-derived neurotrophic factor (GDNF) is a recently identified survival factor for several populations of neurons in the central and peripheral nervous system that also regulates kidney development. To study the roles of GDNF in the regulation of tooth innervation and formation, we analyzed by in situ hybridization the expression patterns of GDNF and its receptors Ret, GDNF family receptor alpha-1 (GFRalpha-1), and GFRalpha-2 from the initiation of first molar formation to the completion of crown morphogenesis. At the time of trigeminal axon ingrowth, GDNF mRNAs were expressed in the mesenchyme around the tooth germ (i.e., target field of the dental innervation), suggesting that it is involved in the regulation of the embryonic tooth innervation. This hypothesis was supported by the ability of GDNF to induce neurite outgrowth from embryonic day 12 (E12) to E15 trigeminal ganglia. This timing correlated with the appearance of Ret in the subset of cells in the trigeminal ganglion at E12, whereas GFRalpha-1 and GFRalpha-2 receptors were constantly expressed in trigeminal ganglion during E11 E15. After birth, GDNF expression showed apparent correlation with the ingrowth and presence of trigeminal nerve fibers in the tooth, suggesting that GDNF is involved in the regulation of innervation of the dental papilla and dentin postnatally. Ret, GFRalpha-1, and GFRalpha-2 mRNAs were expressed in the dental epithelial and mesenchymal cells at stages when epithelial-mesenchymal signalling regulates critical steps of tooth morphogenesis. Ret and GFRalpha-2 were colocalized in the dental mesenchyme during bud and cap stages. Expression of GFRalpha-1 associated with the formation of the epithelial enamel knot, which is a putative embryonic signalling center regulating tooth shape. During postnatal development, GDNF and its receptors were expressed in dental papilla mesenchyme. In addition, GDNF and GFRalpha-1 transcripts were seen in the preodontoblasts and odontoblasts, suggesting that they may be involved in differentiation and maintenance of functional properties of the odontoblasts. Taken together, these results suggest that GDNF acts as a target-derived neurotrophic factor during tooth innervation. In addition, GDNF and its receptors may have nonneuronal organogenetic functions during tooth morphogenesis. PMID- 9415432 TI - Yolk sac-derived murine macrophage cell line has a counterpart during ES cell differentiation. AB - Macrophages are phagocytic hematopoietic cells involved in several immune processes, but they are also present early in mammalian development and may participate in embryonic tissue remodeling. We have isolated and characterized a cell line, Py-YSA, from the mouse yolk sac. Py-YSA cells have several functional properties of macrophages, including uptake of acetylated low density lipoprotein and phagocytic capability. They express the murine macrophage markers F4/80 and Mac-1, and they express RNA for the c-fms receptor. Their expansion in culture requires fibroblast conditioned medium or exogenous monocyte-colony stimulating factor. Murine ES (embryonic stem) cell cultures that undergo in vitro differentiation recapitulate yolk sac development, and during this process cells arise that express both Mac-1 and F4/80 and morphologically resemble the Py-YSA cells. The kinetics and distribution pattern of the Mac-1+ cells during a time course of ES cell differentiation suggest that they originate in the blood islands, and that they subsequently leave the blood islands and disperse to tissue sites. Both F4/80 and Mac-1 are first expressed in primary cultures from day 9.5 yolk sacs. The Py-YSA cultured cells thus resemble embryonic tissue macrophages by several criteria, and they share a marker profile with a cell type found in yolk sacs and differentiating ES cells. Py-YSA cells will be a useful reagent for further understanding the role of embryonic tissue macrophages in development. PMID- 9415434 TI - Can protein unfolding simulate protein folding? PMID- 9415433 TI - Dlx genes encode DNA-binding proteins that are expressed in an overlapping and sequential pattern during basal ganglia differentiation. AB - The Dlx gene family encodes homeodomain proteins that are required for forebrain and craniofacial development. Towards elucidating the roles for each of these genes, we have isolated cDNA clones encoding the full-coding sequence for murine Dlx-5 and partial coding sequence for murine Dlx-6. Three different classes of sense Dlx-5 cDNA clones were characterized, two of which lack the homeobox. We also identified an antisense Dlx-6 transcript. Genomic analysis shows that the Dlx-5 and -6 genes are linked. Biochemical analysis using gel shift assays demonstrate that DLX-1, -2 and -5 have very similar DNA-binding properties. The expression of Dlx-1, -2, -5, -6 and antisense Dlx-1 and -6 was studied in the midgestation mouse brain. We found that the Dlx genes are expressed in overlapping patterns at different stages of differentiation within the primordia of the basal ganglia. Dlx-1 and -2 are expressed in the least mature cells (in the ventricular and subventricular zones). Dlx-5 appears to be co-expressed with Dlx-1 and -2 in the SVZ, but is also expressed in the postmitotic cells of the mantle. Dlx-6 expression is strongest in the mantle. Antisense Dlx-1 and -6 have their highest expression in the SVZ. These results suggest that each of these Dlx genes may have a distinct role in different steps of differentiation in the basal ganglia. PMID- 9415435 TI - Solubility, crystallization and chromatographic properties of macromolecules strongly depend on substances that reduce the ionic strength of the solution. PMID- 9415437 TI - Residue-residue mean-force potentials for protein structure recognition. AB - We present two new sets of energy functions for protein structure recognition, given the primary sequence of amino acids along the polypeptide chain. The first set of potentials is based on the positions of alpha- and the second on positions of beta- and alpha-carbon atoms of amino acid residues. The potentials are derived using a theory of Boltzmann-like statistics of protein structure. The energy terms incorporate both long-range interactions between residues remote along a chain and short-range interactions between near neighbors. Distance dependence is approximated by a piecewise constant function defined on intervals of equal size. The size of the interval is optimized to preserve as much detail as possible without introducing excessive error due to limited statistics. A database of 214 non-homologous proteins was used both for the derivation of the potentials, and for the 'threading' test originally suggested by Hendlich et al. (1990) J. Mol. Biol., 216, 167-180. Special care is taken to avoid systematic error in this test. For threading, we used 100 non-homologous protein chains of 60-205 residues. The energy of each of the native structures was compared with the energy of 43,000 to 19,000 alternative structures generated by threading. Of these 100 native structures, 92 have the lowest energy with alpha-carbon-based potentials and, even more, 98 of these 100 structures, have the lowest energy with the beta- and alpha-carbon based potentials. PMID- 9415436 TI - Fourier-filtered van der Waals contact surfaces: accurate ligand shapes from protein structures. AB - We describe an improved method for determining the shapes and positions of ligand binding sites on proteins by calculating difference contact surfaces of proteins. We report that such calculations may be carried out efficiently by using the principle of the convolution functional operation. Key to this method are (i) use of contact surfaces rather than accessible surfaces, (ii) use of Fourier filtering to smooth binding site features for which the surface features fluctuate too sporadically to correspond with the shape of a true ligand, and (iii) use of Fourier filtering to obtain a simplified intermediate surface to distinguish between non-contiguous adjacent binding sites. This method for determining the shape and location of substrate binding sites has successfully located a number of experimentally observed substrate binding sites for several different ligands bound to several different proteins and it predicts consistent shapes and positions for previously unobserved substrate binding sites. The shapes of the sites calculated by this algorithm are closer to the shapes of the actual ligands than are shapes of similar sites calculated by other, presently available software. We expect that this method shall be of general utility for predicting protein-ligand interactions. PMID- 9415438 TI - Applying experimental data to protein fold prediction with the genetic algorithm. AB - Specific residue interactions as revealed from a few and readily available experiments can be quite important in shaping a protein's tertiary topology by complementing basic and general folding principles. This experimental information is employed in structure prediction (mainchain topology) based on sequence knowledge and the genetic algorithm with its ability to optimize simultaneously many parameters. Examples investigated include the distribution of cysteinyl S-S bonds, protein side-chain ligands to iron-sulfur cages, cofactor-ligands, crosslinks amongst side-chains, and conserved hydrophobic and catalytic residues. Such interactions yield an improvement in the predicted topology (0.4-6.6 A root mean square deviation in the positions of the backbone C alpha-atoms relative to those observed) compared with those resulting from simulations relying only on basic protein folding principles. For several examples the resultant topology depended critically on knowledge of the few and specific interactions such that the relationship between predicted and observed C alpha-positions was near random without their use. The combined methodology (experimental data and the genetic algorithm) should prove helpful in settings where experiment and theory can cooperate in successive steps to elucidate an unknown structure. PMID- 9415439 TI - Modelling protein unfolding: hen egg-white lysozyme. AB - A novel modelling procedure, which rapidly unfolds a protein by enhancing solvent penetration of its core, was used to investigate the unfolding pathway of hen egg white lysozyme. Early on the unfolding pathway there is a dramatic disruption of the tertiary contacts within the protein, which decouples its domains. Subsequently, the helical domain slowly loses its compactness and the helices fluctuate rapidly. The protein then adopts a 'molten globule-like' structure in which the native beta-sheet is essentially intact. The modelled structures have properties similar to those of lysozyme's experimentally characterized partially folded states and provide insight into its complex (un)folding process. The sequence of unfolding events shows how the unfolding pathway of a multidomain protein may be most similar to its fastest, but not necessarily its dominant, folding pathway. PMID- 9415440 TI - Homology modelling of two subtilisin-like proteases from the hyperthermophilic archaea Pyrococcus furiosus and Thermococcus stetteri. AB - The hyperthermophilic archaeon Pyrococcus furiosus produces an extracellular, glycosylated hyperthermostable subtilisin-like serine protease, termed pyrolysin (Voorhorst,W.G.B., Eggen,R.I.L., Geerling,A.C.M., Platteeuw,C., Siezen,R.J. and de Vos,W.M. (1996) J. Biol. Chem., 271, 20426-20431). Based on the pyrolysin coding sequence, a pyrolysin-like gene fragment was cloned and characterized from the extreme thermophilic archaeon Thermococcus stetteri. Like pyrolysin, the deduced sequence of this serine protease, designated stetterlysin, contains a catalytic domain with high homology with other subtilases, allowing homology modelling starting from known crystal structures. Comparison of the predicted three-dimensional models of the catalytic domain of stetterlysin and pyrolysin with the crystal structure of subtilases from mesophilic and thermophilic origin, i.e. subtilisin BPN' and thermitase, and the homology model of subtilisin S41 from psychrophilic origin, led to the identification of features that could be related to protein stabilization. Higher thermostability was found to be correlated with an increased number of residues involved in pairs and networks of charge-charge and aromatic-aromatic interactions. These highly thermostable proteases have several extra surface loops and inserts with a relatively high frequency of aromatic residues and Asn residues. The latter are often present in putative N-glycosylation sites. Results from modelling of known substrates in the substrate-binding region support the broad substrate range and the autocatalytic activation previously suggested for pyrolysin. PMID- 9415441 TI - Autophosphorylation of the catalytic subunit of cAMP-dependent protein kinase in Escherichia coli. AB - When the catalytic (rC) subunit of cAMP-dependent protein kinase (cAPK) is expressed in Escherichia coli, it is autophosphorylated at four sites, Ser10, Ser139, Ser338 and Thr197 (49). Three of these sites, Ser10, Ser338 and Thr197, are also found in the mammalian enzyme. To understand the functional importance of these phosphorylation sites, each was replaced with Ala, Glu or Asp. The expression, solubility and phosphorylation state of each mutant protein was characterized by immunoprecipitation following in vivo labeling with 32Pi. When possible, isoforms were resolved and kinetic properties were measured. The two stable phosphorylation sites in the mammalian enzyme, Ser338 and Thr197, were shown to play different roles. Ser338, which stabilizes a turn near the C terminus, is important for stability. Both rC(S338A) and rC(S338E) were very labile; however, the kinetic properties of rC(S338E) were similar to the wild type catalytic subunit (C-subunit). Ser338 most likely helps to anchor the C terminus to the surface of the small lobe. Thr197 is in the activation loop near the cleft interface. Mutagenesis of T197 caused a significant loss of catalytic activity with increases in Kms for both peptide and MgATP, as well as a small decrease in k(cat) indicating that this phosphate is important for the correct orientation of catalytic residues at the active site. Replacement of Ser139, positioned at the beginning of the E-helix, with Ala had no effect on the kinetic parameters, stability or phosphorylation at the remaining sites. In contrast, mutation of Ser10, located at the beginning of the A-helix, produced mostly insoluble, inactive, unphosphorylated protein, suggesting that this region, though far removed from the active site, is structurally important at least for the expression of soluble phosphoprotein in E.coli. Since the mutation of active site residues as well as deletion mutants generate underphosphorylated proteins, these phosphorylations in E.coli all result from autophosphorylation. PMID- 9415442 TI - Mutational analysis of Escherichia coli elongation factor Tu in search of a role for the N-terminal region. AB - We have mutated lysine 2 and arginine 7 in elongation factor Tu from Escherichia coli separately either to alanine or glutamic acid. The aim of the work was to reveal the possible interactions between the conserved N-terminal part of the molecule, which is rich in basic residues and aminoacyl-tRNA. The enzymatic characterization, comprising GDP and GTP temperature stability assays and measurement of nucleotide dissociation and association rate constants, GTPase activity and aminoacyl-tRNA binding, shows that position 2 is not involved in aminoacyl-tRNA binding, while position 7 is necessary to accomplish this activity. Furthermore, arginine 7 seems to play a role in regulating the binding of GTP. The three-dimensional structure of the ternary complex, EF-Tu:GTP:Phe tRNAPhe, involving Thermus aquaticus EF-Tu and yeast Phe-tRNA(Phe), shows that Arg7 is in a position which permits salt bridge formation with Asp284, thus binding the N-terminus tightly to domain 2. We propose that this interaction is needed for aminoacyl-tRNA binding, and also for completing the structural rearrangement, which takes place when the factor switches from its GDP to its GTP form. PMID- 9415443 TI - Use of protein A gene fusions for the analysis of structure-function relationship of the transactivator protein C of bacteriophage Mu. AB - A sensitive dimerization assay for DNA binding proteins has been developed using gene fusion technology. For this purpose, we have engineered a gene fusion using protein A gene of Staphylococcus aureus and C gene, the late gene transactivator of bacteriophage Mu. The C gene was fused to the 3' end of the gene for protein A to generate an A-C fusion. The overexpressed fusion protein was purified in a single step using immunoglobulin affinity chromatography. Purified fusion protein exhibits DNA binding activity as demonstrated by electrophoretic mobility shift assays. When the fusion protein A-C was mixed with C and analyzed for DNA binding, in addition to C and A-C specific complexes, a single intermediate complex comprising of a heterodimer of C and A-C fusion proteins was observed. Further, the protein A moiety in the fusion protein A-C does not contribute to DNA binding as demonstrated by proteolytic cleavage and circular dichroism (CD) analysis. The assay has also been applied to analyze the DNA binding domain of C protein by generating fusions between protein A and N- and C-terminal deletion mutants of C. The results indicate a role for the region towards the carboxy terminal of the protein in DNA binding. The general applicability of this method is discussed. PMID- 9415444 TI - Mutational analysis of the CD6 ligand binding domain. AB - CD6 belongs to the scavenger receptor cysteine-rich protein superfamily (SRCRSF), which includes a large number of cell surface proteins. The extracellular region of CD6 is composed of three SRCR domains. The membrane proximal SRCR domain of CD6 (CD6D3) specifically binds activated leukocyte cell adhesion molecule (ALCAM), a cell surface protein which is a member of the immunoglobulin superfamily (IgSF). CD6-ligand interactions have been implicated in immune cell adhesion, T cell maturation and the regulation of T cell activation. We tested 13 CD6D3 mutant proteins for binding to ALCAM and a panel of conformationally sensitive anti-CD6D3 monoclonal antibodies (mAbs). CD6D3 residues were classified according to their importance for structural integrity and ligand binding. The results were analyzed in the light of SRCR domain sequence comparison. A number of residues critical for ligand binding or important for structural integrity cluster in the C-terminal region of CD6D3 which is not conserved in other SRCR proteins. PMID- 9415445 TI - Design and construction of a hybrid immunoglobulin domain with properties of both heavy and light chain variable regions. AB - The complementarity-determining regions (CDRs) of a human kappa light chain were replaced with CDRs from a murine gamma-1 heavy chain and, by use of molecular modeling, key heavy chain framework residues were identified and thus included to preserve the native conformation of the heavy chain CDRs. Co-expression of this hybrid human kappa chain (V[HB]C[L]) with a human kappa chain counterpart (V[L]C[L], engineered to contain murine light chain CDRs) resulted in the secretion of high levels of a heterodimeric protein (V[HB]C[L]::V[L]C[L]) termed 'kappabody'. This protein also had equivalent affinity for antigen as the Fab' of the parent murine IgG1. High-level secretion was also observed for the hybrid chain as homodimers (V[HB]C[L]::V[HB]C[L]), which is not observed for chimeric chains consisting of a heavy chain variable region and light chain constant region, i.e. V[H]C[L] homodimers or single chains are not secreted. This indicates that regions within the variable domain, required for secretion of light chains, reside outside of the hypervariable regions (CDRs) and that the heavy chain CDRs and supporting residues do not prevent secretion. These results demonstrate the possibility of designing small, single-domain molecules possessing a given binding activity which may be secreted at high levels from mammalian cells. PMID- 9415446 TI - Improving in vivo folding and stability of a single-chain Fv antibody fragment by loop grafting. AB - The complementary determining regions (CDRs) from the fluorescein-binding antibody 4-4-20, which yields almost no soluble protein in periplasmic expression in Escherichia coli, were transplanted to the framework of the humanized antibody 4D5. The resulting single-chain Fv fragment (scFv) 4D5Flu showed both a dramatic improvement in soluble expression, even at 37 degrees C, and an improved thermodynamic stability. Antigen affinity was maintained upon this engineering by paying attention to crucial framework-CDR contacts. This demonstrates that the use of superior frameworks is a robust strategy to improve the physical properties of scFv fragments. We also report that the grafted version was selected in phage display over several competing variants of the same antibody with identical binding constant but poorer folding or stability properties. The selection required four panning rounds and a temperature of 37 degrees C and we show that the underlying reason for this selection is a higher fraction of phages carrying functional scFv molecules. PMID- 9415447 TI - Recombinant ferritin: modulation of subunit stoichiometry in bacterial expression systems. AB - We describe a strategy for the creation of recombinant ferritin heteropolymers which mimic the natural heterogeneity of this protein. This method entailed the co-expression of cDNA for both ferritin H and ferritin L subunits in a single bacterium using either a bicistronic vector, in which both cDNAs were expressed from the vector, or a dual vector expression strategy, in which each subunit was expressed from a separate compatible plasmid in a single bacterial host. Electron microscopy and sucrose density gradient centrifugation demonstrated that ferritin assembled spontaneously in such bacteria to form catalytically active proteins of the expected size and shape. Isoelectric focusing revealed that protein isolated from any of these bacteria exhibited a restricted heterogeneity in subunit composition. Such multi-subunit recombinant ferritins spontaneously assembled in bacteria may be useful in further studies of ferritin assembly and function. Our results further suggest that varying expression levels is a simple way to alter levels of individual components within a multi-subunit recombinant protein, and that this approach may be of general utility in assessing the contribution of individual components to the function of multi-subunit proteins or protein complexes. PMID- 9415448 TI - Mutagenesis of a flexible loop in streptavidin leads to higher affinity for the Strep-tag II peptide and improved performance in recombinant protein purification. AB - The Strep-tag, an artificial peptide ligand of streptavidin, has gained use as an affinity handle for the purification and detection of recombinant fusion proteins. In an attempt to achieve tighter complexation of the peptide, streptavidin was engineered and the amino acid residues 44-47 in the flexible loop from 44 to 53, which is close to the binding site, were subjected to random mutagenesis. A fusion between alkaline phosphatase and the Strep-tag II sequence, an improved version of the Strep-tag, was constructed as a molecular probe for peptide binding. By means of a filter-sandwich assay, two streptavidin mutants with significantly stronger binding activity for the Strep-tag II were thus identified from a library of Escherichia coli colonies. Both in an ELISA with the alkaline phosphatase fusion and in a fluorescence titration experiment with the synthetic Strep-tag II peptide, which carried an anthraniloyl group as chromophore, their affinities were found to be higher by more than one order of magnitude compared with wild-type streptavidin. The nature of the amino acid exchanges and an enhanced electrophoretic mobility of the streptavidin tetramers suggest an altered loop conformation to be part of the optimized binding mechanism. When one of the streptavidin mutants was immobilized on a chromatographic column it exhibited clearly improved performance in the purification of Strep-tag II fusion proteins, and desthiobiotin turned out to be a suitable reagent for mild competitive elution. PMID- 9415449 TI - Retrospective evaluation of undergraduate medical education by doctors at the end of their residency time in hospitals: consequences for the anatomical curriculum. AB - Reform of the undergraduate medical curriculum, including the debate on which of its parts or subjects are superfluous, is a topic of interest in many countries. On being examined at the end of their specialization period, doctors were asked to grade the relevance of all subjects in the undergraduate curriculum for training to become a medical doctor. The subjects that gained the highest grades were internal medicine, gross anatomy, physical examination course, physiology, and pharmacology. The five subjects graded least relevant were biomathematics, terminology, social medicine, medical physics, and medical chemistry. More than 80% of the doctors expressed interest in special lectures and courses, e.g., in topographic anatomy at the beginning of their postgraduate training. Retrospective evaluations at the end of residency time are helpful "evidence" to be considered in reforming the medical curriculum, and in particular in defining "core" and "optional" parts of the curriculum. The data show that anatomy is graded as highly relevant in the undergraduate medical curriculum by doctors at the end of their postgraduate training. PMID- 9415450 TI - Ultrastructure of Weber's salivary glands of the root of the tongue in the rat. AB - BACKGROUND: The mammalian tongue encompasses several sizeable agglomerations of minor salivary glands. The ultrastructure of the various glands in the body of that organ has already been determined. In contrast, almost nothing is known of the structure of Weber's glands, a collection of salivary glands in the root of the tongue. METHODS: The entire tongue was extirpated from anesthetized adult male rats that had been perfused with fixative and specimens of tissue that included Weber's glands were dissected from the lingual root. These were prepared for electron microscopic examination by conventional means. RESULTS: Weber's glands in the rat are mixed glands, consisting of long mucous tubules that often are capped by serous demilunes. There appear to be no ducts per se, the mucous tubules increasing in caliber and approaching the crypts of the dorsal lingual surface while still retaining their mucous secretory character. As these ducts near the surface, their lining changes to stratified squamous epithelium. The mucous cells are cytologically similar to those in the sublingual glands of the same animal. Mucous droplets undergo vertical fusion to form centrally situated intracellular channels through which mucus is exocytosed, much in the same manner as goblet cells. The serous cells, which have all of the hallmarks of protein secreting cells, probably empty their secretory granules via cellular extensions that directly reach the tubule lumen since there are no intercellular canaliculi between adjacent mucous cells. CONCLUSIONS: The mucus elaborated by Weber's glands undoubtedly aids in swallowing dry food. We postulate that the serous cells in these glands, as in the more anterior von Ebner's glands, might play a role in the mechanism of taste, especially where posteriorly situated, nonlingual taste buds are concerned. PMID- 9415451 TI - Presence of enamel on the incisors of the llama (Lama glama) and alpaca (Lama pacos). AB - Attempts have been made to define the relationships among the South American camelids, the guanaco, llama, alpaca, and vicuna, by comparing the morphology of their incisors. The alpaca has been reported to have an incisor morphology similar to the vicuna, lacking enamel on the lingual surface. The llama and guanaco are said to have enamel on both the labial and lingual surface of their incisor teeth. These comparisons have been based on gross morphological observations and not on histologic analysis. This study was undertaken to determine whether or not alpaca teeth have enamel on the lingual surface. The cross-sectional histologic anatomy of the incisor teeth was compared in two closely related South American camelid species, the llama (Lama glama), and the alpaca (Lama pacos). Thick sections (300 microm) and scanning electron microscopy were the techniques utilized. The mandibular first, second, and third incisors were examined in four llamas and five alpacas. A substantial layer of enamel was present on all surfaces of all llama incisors. The enamel layer on the labial surface of the alpaca incisors closely resembled that found in the llama. The enamel layer on the lingual surface of the alpaca incisors, although greatly reduced, was distinctly present. Alpacas may be more closely related to guanacos and llamas than to vicunas. A histologic study of vicuna incisors would help to better define the relationships of the four camelids. PMID- 9415452 TI - Mononuclear odontoclast participation in tooth resorption: the distribution of nuclei in human odontoclasts. AB - Osteoclasts and odontoclasts have been considered multinucleated giant cells which resorb hard tissue by ruffled borders. Recently, the authors reported the presence of a mononuclear osteoclast and odontoclast with a ruffled border. However, the relative frequency of such cells and the distribution of the number of nuclei including mononuclear cells in them have not been elucidated. Six human deciduous teeth were used in this study. After fixation and decalcification, tartrate-resistant acid phosphatase (TRAP) activity was detected with the azo dye method, and then TRAP-positive cells were observed on resorbing areas of teeth by light microscopy. The cells for investigation were serially sectioned by semithin sections to observe the presence of resorptive lacuna and the number of nuclei. The TRAP activity was detected in both multinucleated and mononuclear odontoclasts from serial semithin sections, and 242 TRAP-positive cells which formed lacunae on dentin were investigated to determine the frequency distribution of the number of nuclei. The mean number of nuclei per cell was 5.3, and median was 4. Only 2.9% of odontoclasts were mononucleus and 93.8% had 10 or fewer nuclei. The majority of odontoclasts forming lacunae on the dentin were cells with 10 or fewer nuclei, and mononuclear odontoclasts participated in human deciduous tooth resorption together with multinucleated ones. PMID- 9415453 TI - Effects of short-term treatment with the bisphosphonates zoledronate and pamidronate on rat bone: a comparative histomorphometric study on the cancellous bone formed before, during, and after treatment. AB - To study the anti-resorptive effects of zoledronate and pamidronate on growing long bones we have performed a histomorphometric analysis of the three regions of the proximal tibial cancellous bone of bone formed before, during, and after drug treatment. Male rats (190-220 g) were treated subcutaneously for 10 days with zoledronate (0.028-2.8 microg/kg) or pamidronate (3.7-370 microg/kg) and sacrificed 5 days later. To delineate the three regions of cancellous bone, and for dynamic bone histomorphometry, calcein and demeclocycline were injected at various times. Both bisphosphonates caused a dose-dependent suppression of cancellous bone turnover and resorption to produce an increase in cancellous bone, but zoledronate was 100 times more potent than pamidronate. The increase in the bone amount and connectivity was more pronounced in the bone formed during treatment where transient bone resorption and normal bone formation led to a positive bone balance. In the bone formed before treatment, inhibition of bone resorption associated with reduced bone formation produced a net gain in amount of bone. Although both bone regions showed a positive bone balance, more bone accumulated in the bone formed during treatment probably because its trabecular bone surface was three times greater. In the primary spongiosa formed after treatment, a moderate increase in the bone amount and connectivity was observed only at the highest dose of both bisphosphonates. The bone formed before, during, and after treatment with bisphosphonates responds differently due to differences in bone architecture, rates of modeling and remodeling, and period of drug exposure. PMID- 9415454 TI - Embryonic, juvenile, and adult development of the toadfish sonic muscle. AB - BACKGROUND: Sonic muscle fibers intrinsic to the swim bladder of the oyster toadfish Opsanus tau proliferate throughout adult life and have an unusual radial morphology: alternating ribbons of sarcoplasmic reticulum (SR) and myofibrils surround a central core of sarcoplasm. Large fibers in adults form multiple cores, fragment, and appear to divide into smaller, more energy efficient units. METHODS: We examined embryonic to adult development of sonic muscle using electron and light microscopy and focused on the incidence of satellite cells (SC). RESULTS: Muscle fibers form late in the larval period from myoblasts, which do not appear to fuse into myotubes, but enlarge and differentiate myofibrils in a single patch. The SR differentiates from the outside inward, separating the myofibrils into bundles of varying thickness, which often exceed the thickness seen in adults. SCs in juveniles and adults have a sparse cytoplasm and a heterochromatic nucleus. The % SC nuclei (SC nuclei/total nuclei) decreases from a high of 88% in larvae to a low of 1% in adults although the adult average is 10%. No embryonic type fibers in the process of differentiating myofibrils were seen in adults. Small immature fibers, which had not yet formed the central core, have a complete radially organized contractile cylinder. CONCLUSIONS: Immature muscle fibers formed embryonically in the larval period have a different morphology from immature fibers in adults, suggesting that splitting rather than SCs is a major source of new fibers in adults. PMID- 9415455 TI - Distribution of fibronectin, type I collagen, type IV collagen, and laminin in the cardiac jelly of the mouse embryonic heart with retinoic acid-induced complete transposition of the great arteries. AB - BACKGROUND: In the mouse model of complete transposition of the great arteries (TGA) produced by all-trans retinoic acid (RA), parietal and septal ridges in the outflow tract (OT) are hypoplastic. At first, these ridges are generated by an expanded cardiac jelly (mainly myocardial basement membrane). Thereafter, endothelial cells delaminate and invade into the adjacent cardiac jelly to form endocardial cushion tissue (formation of cushion ridge). During cushion tissue formation, basement membrane antigens play an important role in the regulation of this endothelial-mesenchymal transformation. METHODS: To examine whether the myocardial basement membrane components are altered in the RA-treated heart OT, immunohistochemistry for fibronectin, type I collagen, type IV collagen, and laminin was carried out in mouse embryonic hearts at 9.5 and 10.5 ED (embryonic day; vaginal plug = day 0) with or without prior exposure to RA. RESULTS: Particulate/fibrillar fibronectin and fibrillar type I collagen were observed in the thick cardiac jelly of the control heart at the onset of mesenchymal formation. In the RA-treated heart, an intermittent patchy staining for fibronectin and a sparse distribution of type I collagen were observed in the thin cardiac jelly. Laminin and type IV collagen were distributed continuously on the basal surface (layer adjacent to the basal plasma membrane) of endocardium and myocardium in both control and RA-treated hearts. CONCLUSIONS: The alterations in the antigens of the myocardial basement membrane (cardiac jelly) may be responsible for the hypoplasticity of parietal and septal ridges that characterizes RA-induced TGA morphology. This may be one of the reasons why mesenchymal cell formation is inhibited in the RA-induced TGA. PMID- 9415456 TI - Immunohistochemical and structural characteristics of the reticular framework of the white pulp and marginal zone in the human spleen. AB - BACKGROUND: The reticular framework of the white pulp (WP) and marginal zone (MZ) consists of reticulum cells and reticulin fibers. The antigenic heterogeneity of the reticular framework is well documented in the mouse and rat spleen. The aim of the present study is to characterize the reticular framework of the WP and MZ of the human spleen. METHODS: Nine surgically resected human spleens were investigated. Five of the nine spleens were perfused. Formalin-fixed materials were embedded in paraffin and serial sections prepared for hematoxylin-eosin, silver staining, and immunohistochemical examination. Electron and immuno electron microscopy were also applied. Using confocal laser scanning microscopy, the reticular framework was analyzed three-dimensionally. RESULTS: The reticulin fibers of the framework were immunostained for type IV collagen in the WP and MZ. The WP was three-dimensionally delimited by the alpha-smooth muscle actin (alpha SMA)-positive reticulum cells. In the WP, the distribution of alpha-SMA-positive reticulum cells formed the reticular framework of the periarteriolar lymphoid sheath (PALS). They also ensheathed the reticulin fibers. Interdigitating cells (IDCs) were scattered throughout the framework. A few IDCs attached to the framework. In the lymph follicle (LF), reticulum cells were not alpha-SMA positive. The mesh of follicular dendritic cells (FDCs) was found in the germinal center. In places, the reticulin fibers were involved in the mesh of the FDCs and covered by the cytoplasm of FDCs. In the MZ, alpha-SMA-positive reticulum cells were arranged in a mesh pattern and ensheathed the fine reticulin fibers. CONCLUSION: The reticular framework of the PALS, LF, and MZ is specialized into heterogeneous components in the human spleen. The heterogeneity of the framework may induce the segregation of T and B lymphocytes. PMID- 9415457 TI - Expression of the smooth-muscle proteins alpha-smooth-muscle actin and calponin, and of the intermediate filament protein desmin are parameters of cardiomyocyte maturation in the prenatal rat heart. AB - BACKGROUND: Coexpression of alpha- and beta-myosin heavy chain (MHC) is a characteristic of the primary myocardial tube. To establish if the smooth-muscle proteins alpha-smooth-muscle actin (alpha-SMA) and calponin, and the intermediate filament protein, desmin, contribute to the specific functional properties of these early cardiomyocytes, we studied their spatiotemporal expression pattern. METHODS: Sections of prenatal and neonatal Wistar rats were stained with antibodies against alpha- and beta-MHC, alpha-SMA, calponin, and desmin. RESULTS: The expression of alpha-SMA and calponin in embryonic cardiomyocytes increases to reach its highest level at ED14. Subsequently, these proteins gradually disappear, beginning in the interventricular septum (IVS) and followed successively by the compact myocardium of the left ventricle, the right ventricle, and the central atrium. Expression of alpha-SMA persists longer in the ventricular conduction system, making it a convenient marker for the ventricular conduction system of the fetal rat. Desmin becomes expressed one day later than alpha-SMA, but also reaches its peak at ED14, suggesting that a relatively high concentration is required to form mature sarcomeres. CONCLUSIONS: The results indicate that alpha-SMA, calponin, and desmin are involved in the myofibrillar development in rat heart. The presence of spatiotemporal differences in the expression of these proteins reveals regional differences in the developmental timing of cardiomyocyte maturation. The maturation process extends from the compact myocardium in the IVS to the left and right ventricular free walls, whereas the atrioventricular junction, the ventricular trabeculae, and developing ventricular conduction system show a relatively slow maturation. Smooth-muscle proteins may contribute to the slow shortening speed that is characteristic of the embryonic myocardium. PMID- 9415458 TI - Anatomical variant of the lateral pectoral nerve innervating the anterior portion of the deltoid muscle: a case report. AB - BACKGROUND: The deltoid muscle is innervated by the axillary nerve. There is no collateral nerve supply described for this muscle. Palsy of the axillary nerve is common in shoulder trauma due to its close relationship to the surgical neck of humerus. METHODS: A dissection of the pectoral and axillary regions of two female cadavers was performed bilaterally for a detailed analysis of the innervation of the deltoid muscle. RESULTS: A branch of the lateral pectoral nerve provided supplemental innervation to the anterior portion of the deltoid muscle bilaterally in both cadavers. CONCLUSION: A branch of the lateral pectoral nerve could provide collateral nerve supply to the deltoid muscle. The frequency of this anatomical variation requires further exploration. PMID- 9415459 TI - Three-dimensional reconstruction of histological sections using modern product design software. AB - BACKGROUND: Computer-based, three-dimensional reconstruction of histological sections is necessary for solving a diversity of questions in morphology and anatomy. Programming software for this purpose is difficult and time-consuming. Therefore, we tested if commercially available product-design software is useful for reconstructing anatomical virtual models. METHODS: We used Alias Wavefront Studio software on a Silicon Graphics workstation. Sections were projected with a newly developed microslide projector on a digitizing board and digitized interactively. Alternatively, pictures of sections were digitized on screen. The resulting set of contours was used as scaffold around which a surface was created manually. RESULTS: Repeated creation of the same object and subsequent measurements gave results that will be sufficiently reliable for many purposes. The visual presentation of objects satisfy highest demands. Little time is needed to reconstruct simple objects, and the time used for complex virtual models is acceptable. Manipulation of objects on screen is nearly in real time and rendering speed is high. CONCLUSIONS: Product-design software packages are a readily available and useful option for anatomists who want to do virtual reconstructions quickly without programming software themselves. PMID- 9415460 TI - Mucosal morphogenesis and cytodifferentiation in fetal porcine small intestine. AB - BACKGROUND: Development of the small intestine is essential for proper nutrition of the fetus and the neonate. This investigation examines the morphogenesis and cytodifferentiation of developing fetal porcine small intestinal mucosa. METHODS: Fetuses were collected from gilts after hysterectomy. Small intestinal segments were removed and processed for light and electron microscopy. RESULTS: Fetal porcine small intestine developed from a simple tube of stratified epithelium to a tube containing villus and intervillus regions of simple columnar epithelium. This development occurred in a proximal to distal direction. By Day 40 of gestation, cytodifferentiation was evident with the presence of goblet cells and enteroendocrine cells in the duodenum. As development progressed, microvilli lengthened and components of the apical endocytic complex (AEC) were observed. By Day 110 of gestation, tubular and vesicular components of the AEC were confined to the jejunum, whereas large lysosomal vacuoles were observed in the distal jejunum. Duodenal epithelium at Day 110 was similar to postnatal epithelium. CONCLUSIONS: The pattern of fetal porcine small intestinal development is similar to that reported for fetal human small intestine. Villus development and cytodifferentiation occur at similar relative times in gestation when compared to the human. These observations support the use of the fetal pig as a model for investigations of human small intestinal development. PMID- 9415461 TI - Spectrum of malignancy and premalignancy in Carney syndrome. AB - Carney syndrome is a rare, autosomal dominant, multi-system disorder comprising 8 well-characterized findings, only 2 of which need be present for a definitive diagnosis. Benign neoplasms are frequent, but malignancies are thought to be uncommon. We have studied a family to clarify the diagnosis and spectrum of clinical manifestations of the syndrome and to develop guidelines for management. The proposita, a 34-year-old woman had classic findings of Carney syndrome, invasive follicular carcinoma of the thyroid gland, Barrett metaplasia of the esophagus, neoplastic colonic polyps, bipolar affective disorder, and atypical mesenchymal neoplasm of the uterine cervix distinct from the myxoid uterine leiomyoma usually seen in this syndrome. Although thyroid gland neoplasm is rare in Carney syndrome, this patient's most aggressive manifestation was her thyroid carcinoma. The diagnosis of Carney syndrome was established in her 9-year-old son and is a probable diagnosis in her 12-year-old daughter. Endocrine manifestations were prominent in the family with at least 9 relatives in 3 generations affected with various endocrine abnormalities. The findings in this family indicate that the spectrum of manifestations in this pleiotropic gene apparently includes a malignant course with premalignant and endocrinologic disorders not previously recognized. PMID- 9415462 TI - Carrier screening for cystic fibrosis: test acceptance and one year follow-up. AB - We identified 124 carriers among 4,879 patients of prenatal care providers in the Rochester region. Six factors were identified that together permitted a correct classification regarding test acceptance for 77.5% of all subjects. For those pregnant, the most influential of these factors was a more accepting attitude toward abortion. As an indication for abortion, cystic fibrosis (CF) ranked between mild and moderate mental retardation. Of the 124 carrier women identified, we obtained 1-year follow-up information on 100. Mean score for CF knowledge at 1 year (77.4 +/- 13.2%), although significantly lower than immediately after counseling (84 +/- 12.4%), was still significantly higher than after detection but before counseling (51.1% +/- 20.7%). Anxiety about having a child with CF significantly declined from 25.8 +/- 8.0 SD immediately after counseling to 18.9 +/- 7.8 at 1 year (Spielberger State Anxiety Scale). Although 15 carriers regretted having been tested, 83% believed that they benefited from testing, 83% would make the same decision to be tested over again, and 79% would recommend testing to a friend. We conclude that, for most women, CF carrier screening accomplished its purpose: most carriers detected came for counseling, had their partners tested, and, if their partners were also carriers, had prenatal diagnosis. The major undesirable outcomes were that many women testing negative did not understand that a negative result did not exclude being a carrier and that three women found to be carriers did not have their partners tested because of anxiety or the unacceptability of pregnancy termination and therefore may not have carefully considered their decision to be tested. Both of these undesirable outcomes could have been avoided by greater attention to pretest patient education by the primary care provider. PMID- 9415463 TI - Palate structure in human holoprosencephaly correlates with the facial malformation and demonstrates a new palatal developmental field. AB - In this study we analyzed palate structure in holoprosencephaly and correlated it with the facial malformations. Eleven human holoprosencephalic fetuses (three cyclopic, two ethmocephalic, one cebocephalic, four with median cleft lip, and one with short philtrum) at 17-23 weeks of gestation and three children (age 2 1/2, 6 and 7 years) with a single central incisor were studied. Photographic and radiographic methods were used. We found that in holoprosencephaly palate structure is abnormal. The severity of this malformation decreases with decreasing severity of facial malformation. Thus, the study shows a close relationship between the facial and the palatal malformation. In all phenotypes the premaxillary area is malformed. From this region, a fan-shaped field along the midpalatal suture is involved in all facial phenotypes, the fan being broadest in cyclopia and narrowest in the short philtrum malformation. A similar fan-shaped field can be discerned in the face, where the broadest fan also indicates the greatest severity with cyclopia, and the narrowest fan the least severe median lip malformation. In the palate field, the anteroposterior furrows seemingly demarcate the field. The findings may be of importance for the future evaluation of palatal malformations in children. PMID- 9415464 TI - Is there an association of Down syndrome and omphalocele? AB - The possible association of Down syndrome (DS) with omphalocele is controversial. We reviewed the 2,979 live births and stillbirths with DS born from 1983 to 1993 in the catchment area of the California Birth Defects Monitoring Program (CBDMP). We observed one infant with both defects, a number that did not differ significantly from what was expected (P < 0.40). We also reviewed the pathological reports of one of us (L.H.H.) from a series of 36 DS fetuses and neonatal deaths; none had an omphalocele. We then reviewed the literature for epidemiological studies of DS and for epidemiological, surgical, prenatal, and familial studies of omphalocele. Possible biases inherent in each type of study were evaluated. The majority of epidemiological studies showed no association of DS with omphalocele. In surgical series, the occasional infant with both defects was more likely to undergo surgery than infants with omphalocele and trisomies 13 and 18 or other severe birth defects. Inclusion of both omphalocele and umbilical hernia in the same ICD-9 code may explain some of the correlations with DS noticed in a few epidemiological studies. In conclusion, our data suggest that trisomy 21 does not predispose the fetus to an increased risk for an omphalocele. PMID- 9415465 TI - Congenital anterolateral bowing of the tibia with ipsilateral polydactyly of the great toe. AB - We report on two cases of congenital unilateral anterolateral bowing and focal defect of the tibia associated with ipsilateral polydactyly of the great toe. Computed tomographic examination showed an unusual partial cleft of the tibia at the site of bowing. A long follow-up of one patient showed spontaneous resolution of the bowing without progression to pseudoarthrosis. These anomalies should be considered as a new entity related to the tibial developmental field. PMID- 9415466 TI - DK phocomelia phenotype (von Voss-Cherstvoy syndrome) caused by somatic mosaicism for del(13q). AB - DK phocomelia (von Voss-Cherstvoy syndrome) is a rare condition characterized by radial ray defects, occipital encephalocoele, and urogenital abnormalities. Lubinsky et al. [1994: Am J Med Genet 52:272-278] pointed out similarities between this and the del(13q) syndrome. To date, all reported cases of DK phocomelia have been apparently normal chromosomally. We report on a case of DK phocomelia in which the proposita had normal lymphocyte chromosomes, but was mosaic in fibroblasts for del(13)(q12). Fibroblast chromosomes studies on other cases of DK phocomelia have not been reported: this raises the possibility that some cases of DK phocomelia may be somatic mosaics for del(13)(q12). PMID- 9415467 TI - Cleft lip and palate, hypertelorism, brachycephaly, flat facial profile, and congenital heart disease in three brothers. AB - A "new" syndrome was identified by McPherson and Clemens [1996: Am J Med Genet 62:58-60] in a brother and sister with bilateral cleft lip and palate, hypertelorism, flat facial profile, flat occiput, and complex heart defects. The brother also had a bilobed tongue and the sister had malrotation of the intestine and bifid thumbs. We describe three brothers with similar anomalies apart from the bilobed tongue, malrotation of the intestine, and bifid thumbs. McPherson and Clemens [1996: Am J Med Genet 62:58-60] suggested autosomal recessive inheritance. Our observation of three affected brothers also raises the possibility of X-linked recessive inheritance. PMID- 9415468 TI - HDR syndrome (hypoparathyroidism, sensorineural deafness, renal dysplasia) associated with del(10)(p13). AB - A combination of hypoparathyroidism, sensorineural deafness, and renal dysplasia has been considered to be a new syndrome inherited in an autosomal dominant fashion; we name the condition "HDR syndrome." We describe a Japanese girl who has HDR syndrome associated with de novo del(10)(p13). The chromosome deletion suggests that the putative gene(s) responsible for HDR syndrome is located at a 10pter-->p13 region. PMID- 9415469 TI - Heterotaxia in a fetus with campomelia, cervical lymphocele, polysplenia, and multicystic dysplastic kidneys: expanding the phenotype of Cumming syndrome. AB - We report on a fetus with tetramelic campomelia, polysplenia, multicystic dysplastic kidneys, and cervical lymphocele. This condition is similar to the autosomal recessive condition described by Cumming et al. [1986: Am J Med Genet 25:783-790] and is different from campomelic syndrome. In addition, our case had anomalies not previously described in this condition, including abnormal lung lobation with bilateral left bronchial morphology, dextrocardia, total anomalous pulmonary venous return, a left superior vena cava, and a right aortic arch. The pancreas was short, with absence of the body and tail. These anomalies are similar to those found in the polyasplenia spectrum. We suggest that the syndrome reported by Cumming et al. may be expanded to include polysplenia with heterotaxia and that Cumming syndrome may be considered another autosomal recessive condition associated with a laterality defect. PMID- 9415470 TI - Disorganization in mice and humans and its relation to sporadic birth defects. AB - Disorganization (Ds) is a mouse mutant best known for producing an exceptional variety of unusual developmental anomalies, such as mirror-limb duplications and hamartomatous skin papillae. So great is the range of malformations that no two affected mice are identical. Several patients with a similar variety of exceptional anomalies have been reported, raising the possibility of the existence of a human homologue of Ds. However, although these human cases represent the most striking findings seen in Ds mice, they do not represent the full range of defects. Most affected mice have only a single malformation, and most of these malformations are similar to both common (neural tube defects, orofacial clefting, gastroschisis, limb reductions) and rare (anophthalmia, duplicated rectum) human birth defects. It is therefore possible that the full spectrum of the human homologue of Ds includes not only patients with the unusual combination of anomalies but also common sporadic birth defects. We suggest that the low penetrance (approximately 0-30%) and highly variable expression of Ds make it a paradigm for understanding the genetic basis for many seemingly sporadic birth defects. PMID- 9415471 TI - Acute idiopathic gastric dilation with gastric necrosis in individuals with Prader-Willi syndrome. AB - Individuals with Prader-Willi syndrome (PWS) have excessive appetite with the ability to consume large quantities of food. Absence of vomiting and a high pain threshold are considered manifestations of the disorder. We present 6 patients with PWS with acute dramatic gastric distention. In 3 young adult women with vomiting and apparent gastroenteritis, clinical course progressed rapidly to massive gastric dilatation with subsequent gastric necrosis. One individual died of overwhelming sepsis and disseminated intravascular coagulation. In 2 children, gastric dilatation resolved spontaneously. Gastrectomy specimens--in 2 cases subtotal and distal, in the other with accompanying partial duodenectomy and pancreatectomy--showed similar changes. All cases demonstrated signs of ischaemic gastroenteritis. All specimens showed diffuse mucosal infarction with multifocal transmural necrosis. Vascular dilatation and small bifrin thrombi were apparent within the infarcted areas. These 6 women with PWS had acute idiopathic gastric dilatation. It is possible that a predisposition to acute gastric dilatation may be related to abnormal gastric homeostasis on a genetic basis. Understanding the mechanisms responsible for this event could increase the understanding of gastrointestinal and appetite regulation in individuals with PWS. PMID- 9415472 TI - Acro-oto-ocular syndrome: further evidence for a new autosomal recessive disorder. AB - We report on a patient born to consanguineous parents and presenting with pseudopapilledema, mixed hearing loss, and minor facial and limb anomalies. To our knowledge, there is just one similar description of this syndrome in three members of a Brazilian kindred whose parents were also consanguineous, suggesting autosomal recessive inheritance. We compare the findings of our patient with these previous reported cases and discuss the differential diagnoses of this new syndrome, which we suggest be named the acro-oto-ocular syndrome. PMID- 9415473 TI - Instability of the FMR2 trinucleotide repeat region associated with expanded FMR1 alleles. AB - The fragile sites FRAXA and FRAXE, located approximately 600 kb apart on Xq27.3 and Xq28, respectively, are due to a CGG trinucleotide repeat expansion. Although the expansion mechanism for these and other trinucleotide repeat disorders remains unknown, the similarities between the FRAXA and FRAXE regions suggest a possible association between the 2 sites. DNA from 953 individuals was analyzed to determine the distribution of FRAXE repeat sizes in this population and to ascertain potential association between FRAXA and FRAXE repeat sizes. Thirty-four FMR2 alleles ranging from 3-42 repeats were identified. No FRAXE expansions were found in this population, supporting previous findings that FRAXE expansions are rare. However, in the fragile X syndrome affected group, a FMR2 delection, 2 cases of FRAXE repeat instability and a FRAXE mosaic male were identified. Also, a previously identified, rare FMR2 polymorphism was observed. Statistical analysis showed no correlation between normal FRAXA and FRAXE repeat sizes studied, although there was a significant size difference in larger FMR2 alleles that segregated with expanded FMR1 alleles. These findings support the idea of an association between repeat expansion in the FMR1 gene and instability or deletions in the FMR2 gene. PMID- 9415474 TI - Urorectal septum malformation sequence: report of thirteen additional cases and review of the literature. AB - We present the findings of 13 additional cases of the urorectal septum malformation (URSM) sequence, and review the literature. The URSM sequence consists of ambiguous genitalia concurrent with absence of perineal and anal openings. The sex ratio of the 13 new cases was 7 males to 6 females and from the literature 21 males and 28 females. In addition, 11 of the 13 new cases had anorectal atresia with 5 of the cases also having partial agenesis of the colon. Bilateral renal agenesis was present in 3 of the 13 cases, unilateral renal agenesis occurred in 6, and dysplastic kidneys were found in 10. The URSM sequence is a lethal condition with long-term survival reported in only 3 of a total of 62 literature and new cases. Recurrence of this condition has not been reported. PMID- 9415475 TI - Population genetics of the FRAXE and FRAXF GCC repeats, and a novel CGG repeat, in Xq28. AB - Most of the rare folate sensitive fragile sites cloned to date arise from expansion of a CGG:CCG trinucleotide repeat array. Analysis of the CAG repeat at the Huntington Disease (HD) locus showed a positively skewed repeat distribution leading to the proposal that microsatellites are subject to a mutational bias toward expansion. Such a mutational bias predicts an increase in mean repeat size at all microsatellite loci. We present an analysis of repeats at two fragile site loci, FRAXE and FRAXF, and a novel CGG repeat in Xq28, in five different human populations, which suggests that these loci may also be subject to the same mutation process. The novel repeat array may represent the first evidence for the existence of a fourth fragile site in Xq27.3-28. PMID- 9415476 TI - Prenatal diagnosis of a familial interchromosomal insertion of Y chromosome heterochromatin. AB - An apparently unbalanced karyotype containing an abnormal chromosome 11 was identified in a 16-week female fetus by analysis of cultured amniocytes. Fluorescence in situ hybridization (FISH) with a chromosome 11 paint identified the presence of an insertion in band 11q24. Parental karyotyping documented an unbalanced karyotype with the same der(11) chromosome in the phenotypically normal father. CBG-banding and FISH identified the insertion to be Yq12 heterochromatin: 46,XY, der(11)ins(11;Y)(q24;q12q12).ish der(11) (wcp11+,DYZ1+). The same der(11) chromosome was also found in the phenotypically normal paternal grandmother, demonstrating this additional Y chromosomal material did not affect normal female sexual development or fertility. The parents elected to continue the pregnancy and a normal girl was born at term, further confirming that this rare familial variant has no clinical significance. This case illustrates the importance of family studies, appropriate banding, and FISH analyses to accurately characterize apparent chromosomal abnormalities. PMID- 9415477 TI - Regional localization of two genes for nonspecific X-linked mental retardation to Xp22.3-p22.2 (MRX49) and Xp11.3-p11.21 (MRX50). AB - Two families with nonspecific X-linked mental retardation (XLMR) are presented. In the first family, MRX49, 5 male patients in 2 generations showed mild to moderate mental retardation. Two-point linkage analysis with 28 polymorphic markers, dispersed over the X-chromosome, yielded a maximal LOD score of 2.107 with markers DXS7107 and DXS8051 at theta = 0.0, localizing the MRX49 gene at Xp22.3-p22.2, between Xpter and marker DXS8022. Multipoint linkage analysis showed negative LOD values over all other regions of the chromosome. In the second family, MRX50, 4 males in 2 generations showed moderate mental retardation. Pairwise linkage analysis with 28 polymorphic markers yielded a LOD score of 2.056 with markers DXS8054, DXS1055, and DXS1204, all at theta = 0.0. Flanking markers were DXS8012 and DXS991, situating the MRX50 gene at Xp11.3 Xp11.21, in the pericentromeric part of the short arm of the X chromosome. PMID- 9415478 TI - Intrafamilial variability of Pfeiffer-type cardiocranial syndrome. AB - We report on the occurrence of Pfeiffer-type cardiocranial syndrome in a brother and sister born to unaffected parents. The cardinal manifestations of the syndrome are congenital heart defect, sagittal craniosynostosis, genital anomalies, and mental and growth retardation. The craniosynostosis was present in only one of the sibs, suggesting intrafamilial variability of the syndrome. The clinical spectrum of cardiocranial syndrome is further expanded by the inclusion of renal, joint, and palpebral abnormalities detected in our patients. The occurrence of the syndrome in a brother-sister sib pair supports the hypothesis of an autosomal-recessive inheritance. PMID- 9415479 TI - Opitz C trigonocephaly syndrome and midline brain anomalies. AB - We describe a child with trigonocephaly, strabismus, upslanting palpebral fissures, nasal bridge hypoplasia, hypertrophic alveolar ridges and large gingivo labial frenula, short neck, hip "dysplasia," equinovarus deformities, cryptorchidism, atrial septal defect ostium secundum, and severe mental retardation, findings consistent with C syndrome. The patient also had a Dandy Walker malformation, complete callosal agenesis, and occipital meningocele. These structural defects are independent of the premature closure of the metopic suture, and confirm that midline brain anomalies are part of C syndrome. The hypothesis that the basic developmental defect in this syndrome primarily affects the midline field is supported by the concomitance of other anomalies, such as conotruncal heart defects, omphalocele, and genital anomalies. PMID- 9415480 TI - Informing parents about the carrier status of their fetus. PMID- 9415481 TI - Lowry-Maclean syndrome with osteopenic bones and possible autosomal dominant inheritance in a Bedouin family. PMID- 9415482 TI - Diastrophic dysplasia sulfate transporter (DTDST) gene is not involved in pseudodiastrophic dysplasia. PMID- 9415483 TI - Further examples of autosomal dominant transmission of nonsyndromic aplasia cutis congenita. PMID- 9415484 TI - Relevance of proto-oncogenes as growth modulators in organogenesis of the mammalian embryonic kidney. PMID- 9415485 TI - Homeobox genes from clusters A and B demonstrate characteristics of temporal colinearity and differential restrictions in spatial expression domains in the branching mouse lung. AB - Lung branching morphogenesis is accomplished by reciprocal morphogenetic interactions between the epithelium and its mesenchyme. In order to better understand the molecular mechanisms regulating these interactions in time and space, the expression patterns of Hox genes isolated exclusively from the branching region of the developing lung have been investigated. Reverse transcriptase PCR identified Hoxa-1, Hoxa-3, Hoxa-5, Hoxb-3, Hoxb-4, Hoxb-6, Hoxb 7, and Hoxb-8 transcripts from within this tissue at 11.5 day post coitum (E11.5). Northern blot, in situ hybridization and PCR analyses demonstrated qualitative and quantitative differences in expression patterns for each gene assessed in this region thus providing evidence for Hox gene temporal colinearity. Furthermore, although not within the context of strict anteroposterior definition, Hox genes located within a more 5' region in both clusters were found to have greater spatial expression constrictions when compared to their more 3' counterparts. These Hox genes were also differentially expressed both between and within specific germ cell lineage derivatives. Such patterns of expression suggest that Hox genes play a role in the specification and maturation of lung cell lineage derivatives throughout the pseudoglandular, canalicular and terminal sac phases of lung development. PMID- 9415486 TI - Characterization and early embryonic expression of a neural specific transcription factor xSOX3 in Xenopus laevis. AB - Using the powerful RDA-PCR-technique we could identify a novel Xenopus specific Sox-gene (xSox3) a transcription factor closely related to the sox sub-group B, which contains a HMG box. In normogenesis the xSox3 gene is expressed in the presumptive central nervous system. Furthermore a maternal component is also found in oocytes and in early cleavage stages in the animal hemisphere only. By whole-mount in situ hybridization the first zygotic transcription activities can be detected in the late blastula in the dorsal ectoderm and the dorsal and lateral part of the marginal zone. The expression reaches the highest level atthe late gastrula till the late neurula and fades after stage 30. The expression is restricted from gastrulation onwards to the presumptive brain area and the lens epithelium. Furthermore we could show that the gene is expressed in isolated Spemann organizer with adjacent neuroectoderm. The signal can be suppressed by suramin treatment, which inhibits neural development and causes a shift of dorsal to ventral mesoderm. The treatment of whole embryos with LiCl and UV results in an overexpression or an inhibition of the expression, respectively. In exogastrulae (pseudo-exogastrulae) the gene is expressed in the close vicinity to the endomesoderm only, but not in the distal most part of the ectoderm. This result indicates that it is unlikely that the gene can be activated by planar signals. The gene can also be activated in dissociated gastrula ectoderm without mesodermal or neural inducers. That means that the gene can be expressed in ectodermal cells in a cell autonomous manner. PMID- 9415487 TI - Mouse molar morphogenesis revisited by three-dimensional reconstruction. III. Spatial distribution of mitoses and apoptoses up to bell-staged first lower molar teeth. AB - Computer-assisted 3D reconstructions were used to follow the development of the embryonic mouse first lower molar (M1). At ED 12.5, the thickening of the oral epithelium, which was thought to correspond to the molar dental lamina, regressed in its anterior part as a result of apoptosis. Only the posterior part later gave rise to molars. The transition to the cap stage entailed medial and lateral extensions of the dental epithelium. The growth and histo-morphogenesis of the enamel organ as well as cervical loop formation proceeded more rapidly in the anterior part of the M1 during the cap and early bell stages producing significant morphological differences along the antero-posterior axis. Apoptosis was temporarily intensive in the anterior part of the bud- and cap-shaped epithelium and thus pointed domains which do not participate in the formation of the final M1 enamel organ. In the well-formed cap, apoptoses displayed maximum concentration in the enamel knot (EK). No increase in the number of metaphases could be detected in the vicinity of the EK. Mitoses were distributed throughout the epithelial compartment until cap stage and then mainly concentrated in the inner dental epithelium at the early bell stage. At this later stage, either lateral views or thick virtual sections performed in the reconstruction demonstrated a clear cut distribution of mitoses and apoptoses in the enamel organ. At the early bell stage, mitoses in the mesenchyme demonstrated an increasing postero-anterior gradient. PMID- 9415488 TI - Isolation of cDNA clones for genes that are expressed in the tail region of the ascidian tailbud embryo. AB - An ascidian tailbud embryo is comprised of the anterior trunk and posterior tail. We constructed cDNA libraries of the tail region and trunk region of the ascidian Halocynthia roretzi. The screening of the tail library by tail single-stranded cDNA minus the trunk library RNA as a probe, yielded cDNA clones for genes that are expressed in the tail epidermis, visceral ganglion, trunk lateral cells, muscle cells, and certain regions of the tail. Among them, a cDNA clone for a gene designated HrPost-1 is described in detail. HrPost-1 encodes a novel, possible secreted protein of 238 amino acids. The expression of the gene is zygotic. HrPost-1 transcript was first evident in the posterior B-line blastomeres including muscle cells and endodermal strand cells of the gastrula stage embryo, and the expression in these regions disappeared by the early tailbud stage. Around neurulation, the HrPost-1 transcript appeared in epidermal cells of the posterior-most region of the embryo. As development proceeded, the gene expression spread anteriorly in the epidermal cells of the neurula and tailbud embryo, and thus at the early-to-mid tailbud stage, HrPost-1 expression appeared to define the boundary between the trunk and tail epidermis. These results suggest that, in addition to tissue-specific genes, the activities of a set of region-specific genes are associated with tail formation in the ascidian embryo. PMID- 9415490 TI - A 3' remote control region is a candidate to modulate Hoxb-8 expression boundaries. AB - Hox genes have been shown to play a key role in the acquisition of positional identity by precursors of embryonic axial, paraxial and limb structures. This function is thought to depend on the sequential, concerted expression of these genes in time and space. However the underlying molecular mechanisms of this collinear expression are still largely unknown. So far we had identified proximal regulatory elements driving expression of Hoxb-8/LacZ transgenes in Hox-like expression patterns with rostral boundaries more posterior than those of the endogenous gene. In this work we have analyzed 30 kb of 3' genomic sequences for Hoxb-8 regulatory activity in transgenic mice. We have identified a control region in the Hoxb-5/b-4 intergenic region that rostrally extends the Hoxb-8/LacZ expression domain into the posterior hindbrain. In combination with the Hoxb-8 minimal promoter, the 3' control region drives transgene expression with boundaries more anterior than those of Hoxb-8 in the neural tube. When combined with a 4.5 kb Hoxb-8 upstream sequence, where essential proximal regulatory sequences are located, the 3' control region drives transgene expression in a domain which seems to correspond to that of the endogenous Hoxb-8. By deletion analysis we have narrowed down to 550bp the regulatory activity interacting with the Hoxb-8 minimal promoter. We discuss the possibility that this remote 3' enhancer, which is the closest regulatory region found in the cluster to rostrally extend Hoxb-8/LacZ expression, could be involved in the regulation of Hoxb-8 and interact with the proximal control elements. PMID- 9415489 TI - Developmental expression of H3.3A variant histone mRNA in mouse. AB - Cloning and characterization of H3.3A variant histone expression has recently been reported to be associated with meiotic development in mouse testis and ovary. Using Northern analysis and in situ hybridization, the pattern of H3.3A expression was studied during the development of different tissues. In addition to the differential expression detected in male and female meiosis, H3.3A was found to be highly expressed in preantral follicles of adult ovaries and in the basal regions of seminiferous epithelium corresponding to spermatogonia. Different patterns of expression were observed in somatic tissues, which also differed with respect to the developmental stage of the tissue. The lowest expression was detected in adult skeletal muscle. High expressions were found in foetal liver and spinal cord. These different expressions might reflect a possible function of H3.3A in cell differentiation as detected in MEL cells. PMID- 9415491 TI - Targeted over-expression of FGF in chick embryos induces formation of ectopic neural cells. AB - Fibroblast growth factors (FGFs) are known to be involved mainly in mesoderm formation in Xenopus embryos but their participation in other inductive mechanisms such as neural induction has not been clearly established and is now under study. Here, we provide evidence that targeted over-expression of members of this family of growth factors in the periphery of full-length primitive streak chick embryos produces the formation of ectopic neural cells that are able to differentiate into neurons. The supernumerary neural plate obtained derives from the epiblast layer of the blastoderm and show signs of neural differentiation 24 h after the application of FGF. We have used cell labeling and have examined the expression of mesodermal markers to ascertain how this expansion of the neural forming region of the epiblast takes place. We conclude that the new neural cells formed are originated in the region of the epiblast fated to be epithelia and that the induction of the ectopic neural tissue is not mediated by an increase, migration or new formation of axial mesoderm. This strongly suggests that FGF is acting directly on epiblast cells, changing their fate from epidermal ectoderm to neural ectoderm. Therefore, our results show that FGF can induce neural ectoderm when acting on still uncommitted cells and, therefore, it is a putative candidate for acting in normal neural induction during development. PMID- 9415492 TI - Distribution of BDNF and trkB mRNA in the otic region of 3.5 and 4.5 day chick embryos as revealed with a combination of in situ hybridization and tract tracing. AB - We have used a recently developed technique which combines fluorescent tract tracing and in situ hybridization to study co-localization of neurotrophin mRNA and neurotrophin receptor mRNA expression simultaneously with the pattern of innervation in the developing chick ear. Efferent and afferent fibersfrom the VII/VIIIth cranial nerves were retrogradely and anterogradely filled with Dextran amines conjugated to Texas red and the brain stem was incubated for 2 hours in tissue culture medium. The tissue was subsequently fixed, sectioned frozen, mounted and subjected to in situ hybridization analysis using probes for brain derived neurotrophic factor (BDNF) and its tyrosine kinase receptor, trkB. The results show that afferent and efferent fibers to the ear innervate areas of the developing otocyst which express BDNF mRNA. We also found that neurons in the stato-acoustic ganglion express high levels of trkB mRNA whereas the subset of facial motor neurons that is efferent to the ear only had no or very low levels of trkB mRNA. From our results we conclude that chicken otic efferent fibers preferentially project to areas with BDNF mRNA expression. The very low levels of trkB mRNA in these motor neurons compared to afferent neurons innervating the same region suggest that other factors, perhaps co-expressed with BDNF, may support efferents. A possible involvement of afferents in guiding efferents to specific areas of the ear is suggested. PMID- 9415494 TI - Contralateral efferent neurons can be detected in the hindbrain outside of rhombomere 4. AB - A group of efferent neurons whose bodies are located contralaterally and extend projections across the ventral midline of the hindbrain is considered as a rhombomere 4-specific characteristic. These neurons contribute to the vestibulo acoustic nerve. At the level of rhombomere 2, a similar kind of efferents have only been described as a result of several experimental manipulations and have been interpreted as being due to rhombomere 2 acquiring rhombomere 4 identity. Here is shown that contralateral efferents can also be detected in rhombomere 2 of normal mouse and chicken hindbrains. These findings indicate that neural processes crossing the midline should not be considered as a rhombomere 4 specific characteristic. They also imply that the formation of the contralateral efferents at different rostro-caudal levels might be under different genetic controls, because Hoxb-1, which is not expressed in rhombomere 2, seems to be essential for their proper formation in rhombomere 4. PMID- 9415493 TI - Shh, Fgf4 and Hoxd gene expression in the mouse limb mutant hypodactyly. AB - The semidominant mouse mutation hypodactyly (Hd), caused by a deletion within the Hoxa13 gene, results in reduced digits; heterozygotes lack digit I in the hindlimb and homozygotes have only one digit on each limb. We investigated expression of Shh and Fgf4 signaling molecules involved in digit specification in mutant limb buds. Shh and Fgf4 are expressed in the posterior part of the limb buds as normal but expression may be slightly prolonged. The extent of digit reduction in hypodactyly is much more severe than in the Hoxa13 deficient mouse and resembles that in the Hoxa13(-/-)/Hoxd13(-/-) double mutant mouse. We found that the pattern of Hoxd13 and Hoxd11 transcripts was not markedly different in the mutant compared with the normal limbs even though the mutant limbs are narrower. Therefore Hoxd genes are transcribed as normal in the mutant. This makes it likely that the severe digit reductions in hypodactyly are caused by interference with Hoxd13 function at the protein level. Similar interactions between mutant and normal HOX gene products have been suggested to occur in the human semidominant disorder, synpolydactyly, caused by mutations in HOXD13. PMID- 9415495 TI - Cortical rotation is required for the correct spatial expression of nr3, sia and gsc in Xenopus embryos. AB - Beta-catenin, a component of the wnt-signal-transduction pathway, is essential for the formation of the dorsal axis in Xenopus laevis embryos. On the dorsal side of the embryo, beta-catenin is translocated into the nuclei via a process linked to cortical rotation. When cortical rotation is blocked by UV-irradiation, nuclear beta-catenin is found in the vegetal pole of the embryo. Here we show that overexpression of beta-catenin in animal cap explants, in the absence of mesoderm induction, is sufficient to activate the expression of genes with dorsalizing activity such as siamois (sia) and nodal-related 3 (nr3) but not goosecoid (gsc). In embryos ventralized by UV-treatment, the expression of the dorsal-specific genes sia, nr3 and gsc is induced at the vegetal pole after the Mid-Blastula-Transition (MBT). While nr3 and sia expression continues in these embryos until gastrula stages, gsc transcription cannot be maintained. We propose that the spatial separation of the expression domains of genes with dorsalizing activities and the prospective mesodermal region results in the loss of dorsal structures in the embryo. The role of cortical rotation is to generate an overlap of the region with dorsal axis-forming activity, indicated by nuclear translocation of beta-catenin, and the prospective mesoderm in the marginal zone to assure the correct positioning of the Spemann organizer. PMID- 9415496 TI - Local origin of cells in FGF-4 - induced outgrowth of amputated chick wing bud stumps. AB - Urodele amphibians are the only vertebrates that can regenerate amputated limbs, even as adults. However, we have previously shown that amputated chick wing bud stumps can be induced to ((regenerate)) and to form a complete set of correctly patterned skeletal elements, following implantation of beads soaked in fibroblast growth factor-4 (FGF-4). We have now performed Dil injection experiments to determine which cells contribute to FGF-4-induced chick wing bud ((regenerates)). We show that the FGF-4-induced outgrowth of the regenerating wing bud stump is comprised of mesenchyme cells that originate from a region within 200 microm of the FGF-4 bead, and that cells proximal to the bead move distally. PMID- 9415497 TI - The generation of yet another myth on the use of narcotics. PMID- 9415498 TI - A critical review of controlled clinical trials for peripheral neuropathic pain and complex regional pain syndromes. AB - The purpose of this review was to identify and analyze the controlled clinical trial data for peripheral neuropathic pain (PNP) and complex regional pain syndromes (CRPS). A total of 72 articles were found, which included 92 controlled drug trials using 48 different treatments. The methods of these studies were critically reviewed and the results summarized and compared. The PNP trial literature gave consistent support (two or more trials) for the analgesic effectiveness of tricyclic antidepressants, intravenous and topical lidocaine, intravenous ketamine, carbamazepine and topical aspirin. There was limited support (one trial) for the analgesic effectiveness of oral, topical and epidural clonidine and for subcutaneous ketamine. The trial data were contradictory for mexiletine, phenytoin, topical capsaicin, oral non-steroidal anti-inflammatory medication, and intravenous morphine. Analysis of the trial methods indicated that mexiletine and intravenous morphine were probably effective analgesics for PNP, while non-steroidals were probably ineffective. Codeine, magnesium chloride, propranolol, lorazepam, and intravenous phentolamine all failed to provide analgesia in single trials. There were no long-term data supporting the analgesic effectiveness of any drug and the etiology of the neuropathy did not predict treatment outcome. Review of the controlled trial literature for CRPS identified several potential problems with current clinical practices. The trial data only gave consistent support for analgesia with corticosteroids, which had long-term effectiveness. There was limited support for the analgesic effectiveness of topical dimethylsulfoxyde (DMSO), epidural clonidine and intravenous regional blocks (IVRBs) with bretylium and ketanserin. The trial data were contradictory for intranasal calcitonin and intravenous phentolamine and analysis of the trial methods indicated that both treatments were probably ineffective for most patients. There were consistent trial data indicating that guanethidine and reserpine IVRBs were ineffective, and limited trial data indicating that droperidol and atropine IVRBs were ineffective. No placebo controlled data were available to evaluated sympathetic ganglion blocks (SGBs) with local anesthetics, surgical sympathectomy, or physical therapy. Only the capsaicin trials presented data which allowed for meta-analysis. This meta-analysis demonstrated a significant capsaicin effect with a pooled odds ratio of 2.35 (95% confidence intervals 1.48, 3.22). The methods scores were higher (P < 0.01) for the PNP trials (66.2 +/- 1.5, n = 66) than the CRPS trials (57.6 +/- 2.9, n = 26). The CRPS trials tended to use less subjects and were less likely to use placebo controls, double-blinding, or perform statistical tests for differences in outcome measures between groups. There was almost no overlap in the controlled trial literature between treatments for PNP and CRPS, and treatments used in both conditions (intravenous phentolamine and epidural clonidine) had similar results. PMID- 9415499 TI - Trigeminal ganglion neuronal activity and glial fibrillary acidic protein immunoreactivity after inferior alveolar nerve crush in the adult rat. AB - Nerve injury to the mandibular division of the trigeminal nerve has been shown to cause satellite cell reactions that extend beyond the mandibular division of the trigeminal ganglion into the maxillary and ophthalmic divisions. The goal of this study was to determine whether any physiological abnormalities correlated with this dispersal of satellite cell reaction. We investigated the electrophysiological and satellite cell glial fibrillary acidic protein immunoreactivity (GFAP-IR) changes that occur within the trigeminal ganglion 3, 10 and 59 days after a crush injury of the inferior alveolar nerve (IAN). At 3 days after IAN crush, there were no mechanically-evoked responses to ipsilateral stimulation of the skin and intraoral structures (e.g., mandibular incisor, lower lip and rostral mandibular gingiva) innervated by the IAN. However, the peripheral representations of the auriculotemporal, mylohyoid, lingual and maxillary nerve were intact and no abnormal responses to mechanical stimulation were detected to stimulation of tissue innervated by these nerves. By 10 days after the IAN crush, mandibular neurons responded to mechanical and electrical stimuli of the peripheral receptive field of the IAN, but with slower conduction velocities and higher electrical thresholds compared to control values. These abnormal electrophysiological response characteristics persisted 59 days following nerve injury. At 3, 10 and 59 days after IAN crush, 3-4% of the recorded mandibular neurons displayed spontaneous activity that was never observed in rats without nerve injury. Spontaneous activity was also never observed in neurons recorded in the maxillary or ophthalmic divisions of the trigeminal ganglion. Intense GFAP-IR in satellite cells was observed surrounding a mean of 131.7 neurons/section within the mandibular division of the trigeminal ganglion 3 days after nerve injury and around 50.3 neurons/section at 10 days. GFAP-IR was also present surrounding 16.5 and 10.3 neurons/section in the maxillary division of the trigeminal nerve at 3 and 10 days, respectively. At 59 days after IAN crush, GFAP-IR satellite cells were found around 22.9 neurons/section in the mandibular division of the trigeminal nerve, but were not found elsewhere in the trigeminal ganglion. The more extensive distribution of neurons encircled by satellite cell GFAP-IR compared to the trigeminal ganglion region containing abnormal electrophysiological responses demonstrates that abnormal neuronal signaling may not be characteristic of trigeminal ganglion neurons that are surrounded by GFAP injury reactions. However, the persistence of GFAP-IR 59 days after nerve injury suggests that satellite cell GFAP is involved in the long-term recovery of injured neurons. PMID- 9415500 TI - The intrinsic antinociceptive effects of oxycodone appear to be kappa-opioid receptor mediated. AB - Our previous studies in the Sprague-Dawley rat showed that the intrinsic antinociceptive effects of oxycodone are naloxone reversible in a manner analogous to morphine but that in contrast to morphine, oxycodone's antinociceptive effects have a rapid onset of maximum effect (approximately 5-7 min compared to 30-45 min for morphine), comprise one antinociceptive phase (compared to two phases) and are of relatively short duration (approximately 90 min compared to approximately 180 min). In the present study, administration of a range of selective opioid receptor antagonists has shown that the intrinsic antinociceptive effects of oxycodone (171 nmol) are not attenuated by i.c.v. administration of (i) naloxonazine, a mu1-selective opioid receptor antagonist, or (ii) naltrindole, a delta-selective opioid receptor antagonist, in doses that completely attenuated the intrinsic antinociceptive effects of equipotent doses of the respective mu- and delta-opioid agonists, morphine and enkephalin-[D Pen(2,5)] (DPDPE). Although beta-funaltrexamine (beta-FNA) attenuated the antinociceptive effects of oxycodone (171 nmol i.c.v.), it also attenuated the antinociceptive effects of morphine and bremazocine (kappa-opioid agonist) indicative of non-selective antagonism. Importantly, the antinociceptive effects of oxycodone (171 nmol i.c.v.) were markedly attenuated by the prior i.c.v. administration of the selective kappa-opioid receptor antagonist, norbinaltorphimine (nor-BNI), in a dose (0.3 nmol) that did not attenuate the antinociceptive effects of an equipotent dose of i.c.v. morphine (78 nmol). Taken together, these data strongly suggest that the intrinsic antinociceptive effects of oxycodone are mediated by kappa-opioid receptors, in contrast to morphine which interacts primarily with mu-opioid receptors. PMID- 9415501 TI - The efficacy of radiofrequency lesioning of the cervical spinal dorsal root ganglion in a double blinded randomized study: no difference between 40 degrees C and 67 degrees C treatments. AB - The efficacy of radiofrequency lesion treatment of the cervical dorsal root ganglion (RF-DRG) in cervicobrachialgia was investigated in 61 patients by a randomized prospective double blinded study. Before lesion treatment the putative pain provoking spinal root was identified by diagnostic blocks with a local anesthetic agent. One group of patients (n = 32, group I) was treated with a radiofrequency lesion of 67 degrees C and in a control group (n = 29, group II) a temperature of 40 degrees C was applied. Three months after treatment a significant reduction in VAS scores was demonstrated in both groups. The outcome of the treatments was identical (VAS reduction: group I, 1.7; group II, 1.9; P = 0.001). In group I a VAS reduction of 3 or more occurred in 11/31 (34%) and in group II in 11/29 (38%) of patients. A VAS reduction of 2 or more occurred in group I in 15/31 (47%) and in group II in 15/29 (51%) of patients. This study suggests that treatment with 40 degrees C radiofrequency application of the dorsal root ganglion is equally effective as treatment at 67 degrees C. Further appraisal of this treatment is required. PMID- 9415502 TI - Pain coping and the pain experience during mammography: a preliminary study. AB - This study examined how pain coping efficacy and pain coping strategies were related to reports of pain during mammography. Subjects were 125 women over the age of 50 undergoing screening mammograms. Prior to their mammogram, all subjects completed the Coping Strategies Questionnaire (CSQ) to assess how they cope with day-to-day pain experiences. Ratings of pain during the mammogram were collected using a 6-point pain/discomfort scale, a 100-mm Visual Analog Scale, the adjective checklist of the McGill Pain Questionnaire, and the Brief Pain Inventory. Up to 93% of the women reported the mammogram examination was painful. On average, women rated the mammography pain in the low to moderate range. Considerable variability in pain ratings was found, however, with some women reporting severe pain and others reporting little or no pain. Correlational analyses were conducted to examine how coping efficacy (CSQ ratings of ability to decrease pain and ability to control pain) and coping strategies (CSQ pain coping strategy subscales) related to variations in pain report. There was a pattern for ratings of ability to decrease pain to be related to lower ratings of current mammography pain. Women who rated their ability to decrease pain as high reported lower average levels of mammography pain, lower ratings on the mammography pain/discomfort scale, and were much more likely to report having had lower levels of pain during their last mammogram. These findings suggest that women who rate their coping efficacy in decreasing day-to-day pain as low may be at higher risk for having a painful mammogram. Individual pain coping strategies were not generally correlated with pain ratings. Behavioral interventions (e.g., patient controlled breast compression) and cognitive therapy interventions (e.g., training in the use of calming self-statements or distraction techniques) designed to increase coping efficacy potentially could be useful in reducing pain in women who are at risk for pain during mammography. PMID- 9415503 TI - Objective evidence of decreased pain perception in normotensives at risk for hypertension. AB - Results from laboratory and naturalistic studies have demonstrated decreased subjective pain ratings in hypertensives and individuals at risk for hypertension. Based on previous evidence that the nociceptive withdrawal reflex may provide an objective index of pain threshold in humans, the present study examined the intensity of sural nerve stimulation required to elicit nociceptive withdrawal in offspring of hypertensives and normotensives. Participants included 60 men and 56 women who were normotensive, 18-23 years of age, and predominately Caucasian. To assess the nociceptive withdrawal reflex, ascending and descending intensities of electrical stimulation were applied over the sural nerve while electromyographic activity was recorded from the ipsilateral biceps femoris muscle. Analyses of the intensity of electrical stimulation required to reach the thresholds for nociceptive withdrawal and subjective pain revealed a pattern of hypoalgesia in individuals at risk for hypertension. First, significantly higher intensities were required to elicit nociceptive withdrawal in offspring of hypertensives versus normotensives. Second, offspring of hypertensives endured significantly more intense stimulation before reporting pain. Third, both parental history of hypertension and resting systolic blood pressure were significant independent predictors of stimulation intensity at nociceptive withdrawal reflex and subjective pain thresholds. These results confirm and extend previous observations of an association between risk for hypertension and hypoalgesia, and suggest that hypoalgesia should be examined as a potential predictor of progressive blood pressure increases in individuals at risk for hypertension. PMID- 9415504 TI - Pain coping and the pain experience in children with juvenile chronic arthritis. AB - This study examined the pain experience and pain coping of children with juvenile chronic arthritis (JCA). The purpose of the study was to describe present pain and the pain coping strategies utilized by children with juvenile chronic arthritis and examine pain coping strategies and pain efficacy as a predictor of pain intensity and distribution. Fifty-six children with JCA rated their present pain using two measures of pain intensity, the Oucher and the pain thermometer, and reported on the number of pain locations using a body map. In addition, each child completed the Child Version of the Coping Strategies Questionnaire (CSQ-C) and health status was determined by a physician's disease activity rating. On average, children reported current pain in the low to middle range on the different pain scales, although there was considerable variability in pain ratings. Up to 30% of all children had pain ratings higher than or equal to the middle range on both the Oucher and the pain thermometer. On average, children reported pain in more than two body areas. Correlational analyses were conducted to examine how the composite factors on the CSQ-C (Pain Control and Rational Thinking, and Coping Attempts) related to variations in reported pain intensity and location. Children who scored higher on the Pain Control and Rational Thinking factor of the CSQ-C had much lower ratings of pain intensity and reported pain in fewer body areas. Hierarchical regression analyses indicated that disease activity and scores on the Pain Control and Rational Thinking factor of the CSQ-C each accounted for a unique, statistically significant proportion of variance in the measures of pain intensity and pain location. Behavioral and cognitive therapy interventions designed to increase pain coping efficacy may be useful adjuncts in treating pain in children with chronic arthritis. PMID- 9415505 TI - Pain coping strategies that predict patients' and spouses' ratings of patients' self-efficacy. AB - This study examined the relationship of pain coping strategies to osteoarthritis patients' ratings of self-efficacy and to spouses' ratings of the patients' self efficacy. Subjects, 130 individuals having osteoarthritis of the knees and persistent knee pain, completed a pain coping strategies measure (the Coping Strategies Questionnaire), a measure of self-efficacy (the Arthritis Self Efficacy Scale), and a measure of pain (the McGill Pain Questionnaire). Two sets of regression analyses were conducted, one examining the degree to which pain coping strategies predicted patients' self-efficacy ratings, and the other examining the degree to which coping strategies predicted spouses' ratings of the patients' self-efficacy. Several pain coping strategies were found to predict a significant proportion of variance in patients' ratings of self-efficacy: (i) ignoring pain sensations was related to higher self-efficacy for pain; (ii) coping self statements were related to higher self-efficacy for controlling other arthritis symptoms (e.g., fatigue or mood symptoms: and (iii) catastrophizing was related to lower self-efficacy for pain, and self-efficacy for other arthritis symptoms. Pain coping strategies were also found to predict a significant proportion of variance in spouses' ratings of the patients' self-efficacy. Specifically: (i) diverting attention was related to lower spousal ratings of self-efficacy for pain; (ii) praying or hoping was related to lower spousal ratings of self-efficacy for function; and (iii) catastrophizing was related to lower spousal ratings of self-efficacy for control of fatigue or mood symptoms. The findings regarding coping strategies were particularly interesting in that they were obtained even after controlling for pain intensity and demographic variables. The pain coping strategies identified are potentially important targets for cognitive-behavioral assessment and treatment efforts. Interventions designed to increase the use of adaptive pain coping strategies and decrease the use of maladaptive pain coping strategies could enhance self-efficacy, reduce pain, and improve the physical and psychological functioning of individuals having osteoarthritis. PMID- 9415506 TI - Peripheral CGRP release as a marker for neurogenic inflammation: a model system for the study of neuropeptide secretion in rat paw skin. AB - The local release of pro-inflammatory neuropeptides in the periphery has been associated with the development of neurogenic inflammation. However, there is an increasing number of reports demonstrating tissue-dependent differences regarding the mechanisms engaged by these neuropeptides to initiate and maintain the inflammatory response in the target tissue. Since skin is often involved in tissue injury, the present studies were designed to develop a model for assessing cutaneous peptide secretion as a marker for neurogenic inflammation in skin tissue. Calcitonin gene-related peptide (CGRP), as one of several neuropeptides known to be involved in neurogenic inflammation, was chosen to study capsaicin induced effects on peripheral neurosecretion. The corial surface of the hairy skin of a rat hindlimb was superfused in vitro, and the basal and capsaicin evoked peripheral release of immunoreactive CGRP (iCGRP) was measured using a radioimmunoassay. The main objectives of these studies were to characterize the various properties of this release including dose-dependency, exocytosis and receptor-mediation as well as the effects of acute and long-term capsaicin desensitization. Capsaicin significantly and dose-dependently increased the release of iCGRP at concentrations ranging from 3 to 300 microM. Omission of calcium ions or treatment with the competitive capsaicin receptor antagonist capsazepine completely inhibited the capsaicin-induced iCGRP release. Superfusion of the skin with 100 microM capsaicin following a conditioning stimulation with capsaicin at concentrations ranging from 0.3 to 100 microM led to an acute, dose dependent desensitization of the CGRP response. In addition, chronic desensitization following the neonatal injection of capsaicin completely abolished the acute iCGRP response to capsaicin. The method described here should prove to be a valuable tool for the evaluation of the processes regulating the peripheral, cutaneous release of pro-inflammatory neuropeptides. This strategy, therefore, may lead to a better understanding of the mechanisms involved in the development and maintenance of neurogenic inflammation, particularly in the skin. PMID- 9415507 TI - Physician and patient factors influencing the treatment of low back pain. AB - Previous retrospective studies have suggested that patient demographics may influence analgesic administration. These studies have not taken physicians' impression of patient pain into account. This prospective study investigates the influence of (i) physician impression of the degree of pain and (ii) patient demographics on the use of analgesic. A convenience sample of adults with non traumatic lower back pain was studied. Possible predictors of analgesic administration included physician pain scores (assessed by visual analogue scale), patient ethnicity, gender, age, and insurance. These variables were tested individually and then using logistic regression. For the total of 91 patients enrolled, only physician pain scale was found to be associated with analgesic use. Median scores were 68 mm (interquartile range = 62-80 mm) for those receiving treatment versus 48 mm (interquartile range = 30-58 mm) for those who did not (P < 0.001). This study therefore suggests that physician impression of patient pain rather than patient demographics influences analgesic use. PMID- 9415508 TI - One-year follow-up of first onset low back pain. AB - Efforts to examine the process and risk of developing chronic back pain have relied generally upon retrospective study of individuals with already established pain. In an alternative approach to understanding the clinical course and evolution of low back disorders, a cohort of 76 men experiencing their first episode of back pain was assessed prospectively at 2, 6 and 12 months following pain onset. Standard measures of pain (Descriptor Differential Scale: DDS), disability (Sickness Impact Profile: SIP), and distress (Beck Depression Inventory: BDI) were employed to classify the sample into five groups: Resolved, Pain Only, Disability/Distress Only, Pain and Mild Disability/Distress, and Clinical Range. At both 6 and 12 months post pain onset, most (78%, 72% respectively) of the sample continued to experience pain. Many also experienced marked disability at 6 months (26%) and 12 months (14%). At 12 months, no participants had worsened relative to the 2-month baseline. Doubly multivariate analyses of variance (MANOVAs) were employed to compare baseline groups (Pain Only, Pain and Mild Disability/Distress, Clinical Range) on the DDS, SIP, and BDI across time. The group by time interaction from 2 through 12 months was reliable, with greatest change occurring in the Clinical Range group in disability and distress; interestingly, the decrease in pain was comparable among all groups. Follow-up tests across measures demonstrated greater change in the early (2-6 month) interval and relative stability in the later (6-12-month) interval. Comparison of those classified as 'improvers' with those who did not improve from 2 to 12 months showed similar findings. The clinical course of first onset back pain may be prolonged for many patients, and involves a continuum of related disability and distress. Individuals at risk for marked symptoms 1 year after an initial episode of back pain can be identified early, and prompt treatment might reduce the risk of pain chronicity. PMID- 9415509 TI - Lamotrigine (lamictal) in refractory trigeminal neuralgia: results from a double blind placebo controlled crossover trial. AB - Lamotrigine is a chemically novel antiepileptic drug which has not been adequately assessed for its antineuralgic properties. It was used in a double blind placebo controlled crossover trial in 14 patients with refractory trigeminal neuralgia. Patients continued to take a steady dose of carbamazepine or phenytoin throughout the trial over a 31-day period. Each arm of the trial lasted 2 weeks with an intervening 3-day washout period. The maintenance dose of lamotrigine was 400 mg. Lamotrigine was superior to placebo (P = 0.011) based on analysis of a composite efficacy index which compared the numbers of patients assigned greater efficacy on lamotrigine with those assigned greater efficacy on placebo. Efficacy for one treatment over another was determined according to a hierarchy of: (i) use of escape medication; (ii) total pain scores; or (iii) global evaluations. Eleven of the 13 patients eligible for inclusion in the composite efficacy index showed better efficacy on lamotrigine compared with placebo. Global evaluations further suggested that patients did better on lamotrigine than placebo (P = 0.025). The adverse reactions with both lamotrigine and placebo were predominantly dose-dependent effects on the central nervous system. A 14th patient withdrew from the study due to severe pain during the placebo arm of the trial. It would appear that lamotrigine has antineuralgic properties. PMID- 9415510 TI - Afferent large fiber polyneuropathy predicts the development of postherpetic neuralgia. AB - Acute zoster infection may be followed by a chronic pain syndrome, i.e., postherpetic neuralgia (PHN). Besides older age, the intensity of pain and neuronal damage within the acutely affected body region are regarded as predictors or risk factors for PHN. As an alternative approach an underlying peripheral polyneuropathy may be considered as potential co-factor. Is a preexisting generalized impairment of certain fiber classes important in initiating chronic pain states after subsequent localized nerve lesion due to zoster infection? Neurophysiological tests of different efferent and afferent small and large fiber systems were performed prospectively at unaffected body regions in patients with acute herpes zoster. Patients that were still in pain 6 months later (PHN, n = 17) and pain free patients (non-PHN, n = 17) were compared regarding the results obtained during the acute phase. Both groups were age matched. Nociceptive C-fiber function was assessed at the forearm by quantitative measurement of the axon reflex vasodilatation and flare induced by histamine iontophoresis. Mechanosensitive A beta-fibers were tested at all extremities by quantitative vibrametry. Parasympathetic small fiber function was studied by heart rate variability tests. No clinically manifest polyneuropathy was present. However, in PHN risk patients considerably higher vibration detection thresholds in hands and feet were detected compared with non-PHN patients. Pathologic test results of vibration sense at the lower extremity predicted PHN with a sensitivity of 70%. Nociceptive C-fiber and parasympathetic fiber function demonstrated no significant differences in both groups. Acute zoster pain was slightly more intense in the PHN group. We concluded that (i) a mild generalized impairment of afferent A beta-fiber function (A beta-polyneuropathy) seems to be an important co-factor in the development of PHN and (ii) impairment of vibration sense, i.e., impairment of afferent A beta-fiber function, may be used as a predictor of PHN. PMID- 9415511 TI - Opiate and H1 antagonist effects on histamine induced pruritus and alloknesis. AB - Itching is a well known side-effect of opiate therapy. To gain insight into the possible contribution of opiate receptors to itching we compared the antipruritic effect of naltrexone (Nemexin), an opiate antagonist, to an H1-receptor antagonist and to placebo. In a double blind cross-over study on 15 healthy volunteers, 25 mg naltrexone or placebo was orally given 60 min prior to a histamine stimulus. In a second, otherwise identical experiment, 10 mg cetirizine, an H1 blocker, or placebo was orally given 12 h before the experiment to the same group of volunteers. Histamine was applied iontophoretically to the forearm skin and the following parameters were assessed thereafter: weal and flare size, itch intensity and the extension of the area of alloknesis ('itchy skin') around the application site. Naltrexone had no effect on the vascular histamine reactions 'weal' and 'flare', whereas cetirizine abolished the weal reactions and greatly diminished the flare reactions. Both naltrexone and cetirizine significantly diminished histamine induced itching. In contrast to placebo and cetirizine, naltrexone abolished alloknesis completely in four of 15 volunteers and in the others alloknesis was greatly reduced after naltrexone. Since vascular reactions to histamine are of peripheral origin, whereas alloknesis depends on central nervous mechanisms, our findings suggest a pronounced centrally mediated action of naltrexone on histamine induced pruritus. PMID- 9415512 TI - The personal constructs of coping with chronic low back pain: is coping a necessary evil? AB - The construct of coping is explored in this paper utilising repertory grid technique with a small group of non-patients with chronic pain. Nineteen volunteers with low back pain completed a repertory grid with eight given elements signifying various self and illness-related roles. Two constructs were given and the remainder elicited using the triad method. The 19 participants rated themselves as being in less pain than those they typified as ill or disabled and considered themselves to be coping with their pain. The constructs elicited emphasised authenticity, the limitations of being a coper, mastery, active stoicism, cheerfulness, acceptance and maintaining acceptable social interactions and appearances. Copers were considered to not be in constant pain. Self, ideal-self and social-self constructs were closely related. The participants rated themselves more like copers than ill, pain-suffering, invalid or hypochondriacal persons. Being a coper, however, was less desirable than being pain free. In essence, these volunteers with low back pain see coping as a necessary evil. This ambivalent and ambiguous construing of coping needs to be further explored in community and patient groups if we are to improve the collaboration between patients and therapists in achieving good pain management. PMID- 9415513 TI - Determinants of pressure pain threshold in adult twins: evidence that shared environmental influences predominate. AB - The objective of this study was to examine the relative contribution of genetic and environmental factors in determining pain perception in a classical twin study. Dolorimeter measurements of pressure pain threshold (PPT) were recorded in 609 healthy female-female twin pairs of whom 269 pairs were monozygotic (MZ) and 340 were dizygotic (DZ). There was a strong correlation (R) in PPT in both MZ and DZ pairs (R(MZ) = 0.57, 95% confidence interval (CI): [0.49, 0.65]; R(DZ) = 0.51, 95% CI: [0.42, 0.59]). The slight excess in intraclass correlation observed in MZ when compared with DZ twins corresponds to a heritability for PPT of only 10% and is not statistically significant. Neither estimate of intraclass correlation was substantially altered after adjusting for a range of potential confounding variables including age, current tobacco and alcohol use, current analgesic use, psychological status assessed by the general health questionnaire, and social class. The dolorimeter measurements were shown to be reliable (between observer agreement R = 0.66; within observer agreement R = 0.70-0.76) and stable over time. In conclusion, these data suggest that there is no significant genetic contribution to the strong correlation in PPT that is observed in twin pairs. These findings reinforce the view that learned patterns of behaviour within families are an important determinant of perceived sensitivity to pain. PMID- 9415514 TI - Intravenous high-dose methadone administered by patient controlled analgesia and continuous infusion for the treatment of cancer pain refractory to high-dose morphine. AB - The management of severe tumor-related pain in the patient with cancer may be problematic. Systemically administered opioids remain the cornerstone of treatment for moderate to severe cancer pain, while parenteral routes should be considered for patients who require rapid onset of analgesia, and for highly tolerant patients whose dose requirements cannot be conveniently administered. The use of intravenous methadone by patient controlled analgesia (PCA) is attractive for the management of severe, intractable cancer pain and may offer some advantages over morphine. We describe the safe and effective use of high dose intravenous methadone by PCA and continuous infusion for a patient with intractable tumor-associated cancer pain who experienced inadequate pain control and dose-limiting side-effects with high-dose intravenous morphine. PMID- 9415515 TI - Intravenous phentolamine mesylate alleviates the pain of pancreatic carcinoma. AB - This case report describes the successful alleviation of the pain of pancreatic carcinoma with intravenous phentolamine mesylate. A 2 day infusion gave relief for 26 days on the first occasion and for 12 weeks after the second infusion. The possible mode of action is discussed. PMID- 9415516 TI - Transvaginal amniotic puncture for cytogenetic evaluation of missed abortions. AB - Transvaginal amniotic puncture (TAP) was performed on 20 consecutive missed abortions immediately prior to dilatation and evacuation and the cytogenetic results compared. The information received from products of conception (POC) and TAP was in concordance in only 5 of 20 (25%) cases. Tissue obtained from POC yielded cells in all instances. However, only 3 of 20 POC samples yielded findings other than normal female. In contrast, 92.8% of the conclusive diagnoses would have been achieved by TAP alone. These data strongly suggest that TAP is superior to POC for accurate cytogenetic assessment of missed abortion and should lead to a reevaluation of our current understanding and management of pregnancy loss. PMID- 9415517 TI - Messenger RNA expression of endothelin-1, endothelin-A receptor and endothelial constitutive nitric oxide synthase in hydatididorm moles. AB - An interesting feature in molar pregnancy is the association with preeclampsia. The reason for this has not been explained but could possibly be due to differences in vasoactive agents compared to normal pregnancy. The aim of this study was to examine the mRNA expression of the vasoconstrictor endothelin-1 (ET 1), its receptor ET-A and the endothelial constitutive nitric oxide synthase (ecNOS), forming the vasodilator nitric oxide, in hydatidiform moles. The results demonstrated the presence of mRNA expression of ET-1, ET-A and ecNOS in hydatidiform moles and that the level of mRNA expression did not vary from that in control placentas. Thus, the present data could not explain the increased frequency of preeclampsia in molar pregnancy. PMID- 9415518 TI - Fetal fibronectin in vaginal fluid of women in prolonged pregnancy. AB - The aim of this study was to determine fetal fibronectin in vaginal fluid of women in prolonged pregnancy, its relationship to a modified Bishop score and its predictiveness of delivery within 3 days. Vaginal samples were collected from 80 women at 42 weeks of gestation for the fetal fibronectin assay. A modified Bishop score was estimated. Fetal fibronectin was determined by a quantitative enzyme immunoassay. The concentration of fetal fibronectin in vaginal fluid was elevated in only 36 of the 80 women. The Bishop score and the time between sampling and delivery were not associated with an elevated fetal fibronectin (> or = 0.05 mg/l). We conclude that fetal fibronectin is not a good indicator of delivery within 3 days. The findings add to our understanding of the complexity of the etiology of postterm labor. PMID- 9415519 TI - Lipid peroxidation in cord blood at birth: a marker of fetal hypoxia during labour. AB - OBJECTIVE: This prospective study examined purine metabolism in relation to free oxygen radical activity, as reflected by lipid peroxide levels in umbilical cord blood at birth. SETTING: Departments of Obstetrics and Gynaecology and of Chemical Pathology, the Chinese University of Hong Kong, Hong Kong, and Purine Research Laboratory, UMDS of Guy's and St. Thomas' Hospitals, London, UK. METHODS: Umbilical cord arterial and venous blood samples were collected from 132 singleton term deliveries for determination of hypoxanthine, xanthine, inosine, uric acid, organic hydroperoxides (OHP) and malondialdehyde. Oxygen saturation, PO2, pCO2, pH, and base excess (BE) were also measured. RESULTS: There was a significant correlation between umbilical arterial and venous levels of hypoxanthine, xanthine, inosine, uric acid and all acid-base parameters (p < 0.001). Significant arteriovenous differences were observed for all parameters with the exception of inosine, uric acid and OHP. Umbilical arterial xanthine and potassium correlated significantly with OHP, but hypoxanthine, inosine and uric acid did not. In 13 babies classified as severely asphyxiated at birth (umbilical arterial pH <7.15, BE <-8), xanthine and OHP levels were significantly elevated when compared with non-asphyxiated babies. No significant differences were observed for hypoxanthine, inosine or uric acid. CONCLUSION: The findings indicate that OHP, either in cord arterial or venous blood, is the best marker of free oxygen radical activity in the fetus, and that this correlates with other evidence of cellular hypoxia-reperfusion injury. We propose OHP is a better measure of perinatal outcome than either acid-base balance or hypoxanthine. PMID- 9415520 TI - Longitudinal measurement of amniotic fluid index in term pregnancies and its association with intrapartum fetal distress. AB - OBJECTIVE: The aim of the present study was to evaluate the dynamic changes in serially obtained amniotic fluid index values and to determine any association with intrapartum fetal distress in a term population. MATERIALS AND METHODS: All patients, > or = 40 weeks of gestational age, evaluated at the Institute of Obstetrics and Gynecology, 'G. Salesi' Hospital, University of Ancona, between January 1, 1994, and December 31, 1995, participated in this longitudinal study. Women with an amniotic fluid index of > 50 mm, who also demonstrated a reactive nonstress test, underwent semiweekly amniotic fluid assessment until spontaneous labor. After 42 gestational weeks, the patients underwent an elective induction of labor. All patients were managed with continuous electronic fetal heart rate monitoring throughout labor. The incidence of intrapartum fetal distress, and meconium staining of amniotic fluid were evaluated with respect to the amniotic fluid index. RESULTS: Of the 117 patients that were evaluated by ultrasound, 83 women had multiple amniotic fluid index measurements and were enrolled in the study. A serial decrease in amniotic fluid index was documented in 54 women; the mean decrease per week was 20.7 +/- 15.4%. An increase in amniotic fluid index was noted in 17, while 11 women showed no change in amniotic fluid index over time. The 14 patients who underwent cesarean section for fetal distress had a significantly lower amniotic fluid index (p < 0.001) at the last sonographic examination than the normal outcome group. Significant differences were also observed for a serial decrease in the amniotic fluid index within a week (p < 0.001). The sensitivity and specificity of the 30% serial decrease in the amniotic fluid index cutoff point, with respect to intrapartum fetal distress were 86 and 93%, respectively. CONCLUSION: Longitudinal measurement of the amniotic fluid index seems to be an effective method in predicting intrapartum fetal distress in a term population. PMID- 9415521 TI - Semen parameters and conception rates after intraperitoneal insemination. AB - OBJECTIVE: To analyze the reproductive outcome in infertile couples which underwent intraperitoneal insemination (IPI). METHODS: We analyzed a series of 216 couples who underwent IPI. Indications for treatment were unexplained infertility in 51 couples and male factor in 165. The 51 couples with unexplained infertility underwent a total of 71 cycles (20 couples underwent a second IPI cycle). The 165 couples with male factor underwent 243 cycles (165 first cycles and 78 second cycles). RESULTS: Out of the 314 cycles performed, a total of 41 clinical pregnancies were observed, with a corresponding conception rate of 13.1%. The values of conception rates for unexplained and male factor infertility were 21.1 (based on 15 pregnancies) and 10.7% (based on 26 pregnancies), respectively. Out of the 41 pregnant women, 26 gave birth to a child, thus the overall livebirth rate was 8.3% (12.7 and 7.0%, respectively, for unexplained infertility and male factor diagnostic subgroups). Considering the couples with unexplained infertility, the conception rates were 9.0, 30.8 and 20.0% for strata of < 5, 5-10 and > or = 10 millions of inseminated spermatozoa. The corresponding values were 6.8, 12.5 and 18.5%, respectively, in couples with male factor infertility (chi(2)1 trend p < 0.05). CONCLUSION: In conclusion, this series provides quantitative estimates of pregnancy rates after IPI in Italian couples with unexplained infertility or male infertility and suggested that the number of motile sperm inseminated is a determinant of pregnancy with this technique. PMID- 9415522 TI - Computed sonography: requiem to echogenicity assessment? AB - We studied the influence of changes in gain settings, log compression, persistence, preprocessing, and postprocessing on image density in the fetal liver model. Each parameter was studied while the others were held constant. The image density was objectively measured by electrooptical transmission densitometry using a transparent film output. Neither the persistence nor the preprocessing levels significantly changed image density. Postprocessing of sonographic images produced significant differences (p < 0.02) in mean image density of most of the various postprocessing curves and is a serious confounder of tissue echogenicity assessment. The data reconfirmed that there is a linear relationship (r = -0.94 to -0.997) between image density and gain setting. However, each log compression setting significantly changed (p < 0.0000001) this relationship, obviating possible image density calibration. Our data suggest that manipulation of image parameters by computed sonographic technology obviates accurate echogenicity assessment. PMID- 9415524 TI - Clinical evaluation of a new model of a transcutaneous electrical nerve stimulation device for the management of primary dysmenorrhea. AB - Transcutaneous electrical nerve stimulation (TENS) has been proven effective in pain relief of primary dysmenorrhea (PD). We evaluated the efficacy of a new TENS device (Freelady, Life Care, Tiberias, Israel), designed to correct disadvantages of older models used in previous studies, in 102 nulliparous women with PD, who were treated with various types of pain relief medications. Marked pain relief was reported by 58 patients (56.9%) and moderate relief by 31 (30.4%). These subjective findings were supported by the fact that the same number of patients (58 and 31) either stopped analgesic use altogether during the trial or reduced the quantity of analgesics, respectively. The device examined proved to be efficient and safe in controlling the pain and disability caused by PD. PMID- 9415523 TI - Prophylactic transabdominal amnioinfusion in oligohydramnios for preterm premature rupture of membranes: increase of amniotic fluid index during latency period. AB - OBJECTIVE: This study was designed to: (i) evaluate the effect of amnioinfusion on the latency period in patients with oligohydramnios for preterm premature rupture of membranes, and (ii) to investigate the relationship between changes in the amniotic fluid index and fetal heart rate short-term variability by computerized Hewlett-Packard cardiotocography, longitudinally estimated before and after prophylactic amnioinfusion. MATERIALS AND METHODS: All singleton pregnancies with prolonged premature rupture of membranes after 25 weeks of gestation and seen at the Institute of Obstetrics and Gynecology, University of Ancona (Italy), between January 1994 and June 1995 were included in the study. Transabdominal amnioinfusion with 150-350 ml warmed normal saline (25-50 ml/min) was performed at weekly intervals. Amniotic fluid volume was assessed ultrasonographically by means of the four-quadrant technique on a weekly basis before and after each amnioinfusion, as well as the short-term variability by a Hewlett-Packard computerized cardiotocographic system. RESULTS: 18 women were enrolled and underwent prophylactic transabdominal amnioinfusion at weekly intervals until delivery. Eighteen controls, who did not undergo prophylactic amnioinfusion, were recruited from our 1992-1993 series and included in the study. The median interval between premature rupture of membranes and delivery was 3.0 weeks (range 1-8 weeks), with an average delivery age of 33.0 weeks (range 27-36 weeks). The latency period was significantly longer in patients who underwent prophylactic amnioinfusion (mean +/- SD, 4.1 +/- 1.7 weeks) than in controls(1.7 +/- 1.0 weeks; p < 0.001). An increase in both the weekly amniotic fluid index (linear regression analysis r = 0.8, p = 0.03) and the weekly short term variability (linear regression analysis r = 0.82, p = 0.02) was observed among patients who underwent prophylactic amnioinfusion. A direct relationship was observed between the amniotic fluid index and short-term variability (linear regression analysis r = 0.54, p = 0.04). The mean values of fetal movements recorded by computerized tomography during the 20 min of observation significantly increased after amnioinfusion in comparison with those before it (2.6 +/- 0.9 and 0.9 +/- 0.7 respectively; p = 0.001). CONCLUSION: The present study has shown a positive effect of prophylactic transabdominal amnioinfusion on the latency period in patients with preterm premature rupture of membranes and oligohydramnios. Among the patients who underwent amnioinfusion, an interesting improvement in fetal heart rate short-term variability was associated with the progressive increase in amniotic fluid volume, as an expression of fetal well being. PMID- 9415525 TI - A clinical method for testing the safety of catamenial pads. AB - Methods are described for assessing the gynecologic, dermatologic, and microbiologic effects of deodorant and non-deodorant catamenial pad use over a 6 month period. A controlled, randomized, investigator-blind parallel study was conducted with 190 women between the ages of 18 and 45 years. Data on medical histories, physical examinations, diagnostic laboratory tests, gynecologic and dermatologic examinations and microbiology were collected. No significant differences in gynecologic, dermatologic, or microbiologic parameters were observed between control and treated groups, and no pad-related adverse health effects were observed in this clinical study. PMID- 9415526 TI - Study of the uterine response to vaginal distension: the 'vagino-uterine reflex'. AB - To study the effect of vaginal distension on the uterus, the uterine pressure was measured in 20 healthy female volunteers (mean age 34.3 +/- 7.8 years; 7 nulli-, 13 multiparous) by means of a manometric tube perfused by a pneumohydraulic system. Vaginal distension was induced by a 12 F condom-ended catheter. The condom was inflated in increments of 10 ml of air. The test was repeated after anesthetizing the vagina and uterus, respectively. The uterine pressure increased upon vaginal distension; it showed more rise as the distension increased (p < 0.01). The response was momentary and the pressure returned to basal values although vaginal distension continued. It disappeared upon sustained or successive inflations of 5-7 times and was restored after a resting period of 3-5 min. There was no uterine pressure response when the vagina or uterus were anesthetized and it returned when the anesthetic effect had worn off. No significant difference was found in the pressure response between nulli- and multiparous women (p > 0.05). The aforementioned results were reproducible when repeated in the same woman with no significant difference (p > 0.05). The inflated condom looks like the erect penis, and the uterine response to the inflated condom seems to simulate that of the erect penis distending the vagina during coitus. The constant rise of uterine pressure to vaginal distension postulates a reflex relation which we call 'vagino-uterine' reflex. Uterine contractions during coitus are suggested to have a 'suction-pumping' action on the semen deposited in the vaginal fornices. PMID- 9415527 TI - Distribution of platinum in human gynecologic tissues and pelvic lymph nodes after administration of cisplatin. AB - BACKGROUND: Defining tissue accumulation of platinum may be of importance, since it may provide a pharmacological explanation for organ-specific cisplatin activity. This study was conducted to evaluate the efficacy of cisplatin at the tissue level in different gynecologic organs. The doses administered were equivalent to those used in neoadjuvant chemotherapy regimens. STUDY DESIGN: Cisplatin was administered intravenously to patients with cervical or endometrial cancer 1 h before operation, and platinum accumulations in tissues were assayed by the atomic absorption method. RESULTS: Platinum accumulation was highest in the cervix and next highest in the myometrium in both cancers. Platinum accumulation in ovary and lymph node was only 0.58 and 0.57 times that in the myometrium, respectively. In patients with cervical cancer, the platinum accumulations in the myometrium and cervix were significantly higher than in the ovary and lymph node. Platinum accumulation in cervical cancer tissue was lower than in the myometrium and cervix, suggesting that delivery of cisplatin to a cervical cancer is somewhat more difficult than to the normal cervix. In patients with endometrial cancer, there was significantly more accumulation in the cervix than in the ovary and lymph node. CONCLUSIONS: These data indicated that cisplatin was easily distributed to the cervix and myometrium, but not to the ovary, lymph node, and cancer tissues. PMID- 9415528 TI - Leiomyomas of the uterus, ovary and vaginal wall. A case report. PMID- 9415529 TI - Squamous carcinoma in situ of the ovary. AB - We report a case of a squamous cell carcinoma in situ of the ovary in a patient previously submitted to radical hysterectomy and pelvic lymphadenectomy for an epidermoid carcinoma of the uterine cervix. The histogenesis of epidermoid tumors of the ovary and their association with squamous malignancies of the uterine cervix are discussed. PMID- 9415530 TI - Gene expression of IL-10 in relationship to TNF-alpha, IL-1beta and IL-2 in the rat brain following middle cerebral artery occlusion. AB - To systematically elucidate the gene expression of inflammatory and immune modulators following middle cerebral artery occlusion (MCAO) in the rat, we studied interleukin-10 (IL-10) along with tumor necrosis factor alpha (TNF alpha), interleukin-1 beta (IL-1beta) and interleukin-2 (IL-2). Gene expression of these cytokines was studied ipsilateral and contralateral to the MCAO, with mRNA expression levels evaluated 2, 4, 6, 8 and 12 h following permanent MCAO by reverse transcriptase polymerase chain reaction (RT-PCR). In the ischemic hemisphere TNF-alpha and IL-1beta mRNA increased at 2 h following MCAO and peaked at 6 h, with IL-10 mRNA detected only at 6 h. Contralaterally, both TNF-alpha and IL-1beta mRNAs were expressed with a similar pattern to that in the ischemic hemisphere, but at lower levels, with no contralateral IL-10 expression. There was no difference in IL-2 gene expression between control and experimental animals in either hemisphere. These results demonstrate that IL-10 and TNF-alpha, IL-1beta gene expression is induced early following MCAO. The temporal profile of these cytokines is similar to that seen in sepsis, where TNF-alpha induces IL-10; subsequently IL-10 inhibits TNF-alpha expression. The similarity of the temporal profile of cytokine expression in sepsis and cerebral ischemia suggests that IL 10 should be studied as a potential inhibitor of TNF-alpha production in ischemic brain tissue. The factors inducing contralateral expression of the inflammatory cytokines, TNF-alpha and IL-1beta, along with the potential clinical significance of this remote cytokine gene expression, merit further study. PMID- 9415531 TI - Chronic administration of suramin induces neurotoxicity in rats. AB - In the present study, the ability of suramin 18 mg/kg i.p. twice a week to induce chronic neurotoxicity in rats was investigated. After 20 weeks of suramin treatment, morphological analysis of nerve fibers demonstrated that 57.7+/-3.2% of them presented vesicular disruption of myelin sheaths; their thickness was 0.23+/-0.07 microm in suramin-treated rats with respect to 0.43+/-0.07 microm of controls (P<0.05). To investigate the interaction between suramin and nerve tissue, the binding of the drug to partially purified myelin P0 protein obtained from sciatic nerves was analysed. The percentage of suramin bound to rat myelin P0 protein was 94.0+/-9.5%; this value was decreased to 55.0+/-7.6% when heparan sulfate was added to the myelin protein suspension before suramin. The analysis of tissue drug concentrations at 5, 10 and 20 weeks of treatment showed that suramin accumulated into the sciatic nerve in a time-dependent fashion (130.8+/ 18.1, 219.7+/-17.1 and 449.3+/-15.6 microg/g of tissue, respectively). In conclusion, suramin induces a chronic peripheral neurotoxicity in rats characterized by myelin damage and high tissue levels of the drug. The high affinity of suramin for partially purified myelin P0 protein suggests a possible mechanism for drug-induced toxicity. PMID- 9415532 TI - The mechanism of action of botulinum toxin type A in focal dystonia is most probably through its dual effect on efferent (motor) and afferent pathways at the injected site. AB - OBJECTIVE: To highlight some clinical and physiological features related to treatment with botulinum toxin type A (BTX-A) injections for focal dystonia that may suggest an effect through efferent (alpha motoneuron) and afferent pathways. DATA SOURCES: This review is based on published clinical and physiological studies as well as personal experience regarding the effect of BTX-A in focal dystonia. DATA SYNTHESIS: Long or short lag period between BTX-A injections and clinical improvement, remote effect, an effect on the basic physiological characteristics of dystonia, poor correlation between the local weakness and the clinical improvement and alleviation of pain are clinical observations which are difficult to explain on the basis of the known effect of BTX-A on the neuromuscular junction of the alpha motoneuron. These observations as well as recent scientific reports are used to discuss a hypothesis that in addition to its effect as local muscle relaxant, BTX-A acts at the level of the central nervous system (CNS) for 'reorganization'. Such an effect on CNS activity can be mediated through afferent pathways coming from the injected site--possibly originated in muscle spindles. Its effect through afferent pathways on the CNS may be considered as a long-term 'sensory trick'. PMID- 9415533 TI - Alpha1-antichymotrypsin polymorphism in Japanese cases of Alzheimer's disease. AB - We examined the possible involvement of alpha1-antichymotrypsin (ACT) polymorphism in the risk for Alzheimer's disease (AD) in a Japanese population. No differences between AD and control subjects have been shown in the genotype distributions and allele frequencies of ACT. No modification of the risk for AD was observed, either alone or in combination with the apolipoprotein epsilon4 (ApoE-4) allele. Our results from a Japanese population failed to confirm the previous data in which the ACT polymorphism was shown to affect the ApoE-4 associated risk for AD. PMID- 9415534 TI - Epidemiology and prognosis of acute myelitis in Southern Finland. AB - In this study we analyzed all acute adult (>15 years) myelitis cases in the province of Uusimaa in Southern Finland during the years 1981-1993. Only cases with acute infectious myelitis were included. Demyelinating diseases, and medullopaties due to degeneration, traumatic, toxic, hereditary, nutritional or metabolic causes were excluded. A total of 45 patients fulfilled the criteria. The mean incidence was 3.5 cases/million inhabitants/year. The mean latency time from the initial infection to the beginning of neurological symptoms was 11 days. Motor paraparesis was found in 62% and tetraparesis in 13%. Sensory symptoms were found in 82% and bowel disturbances were experienced by 71% of patients. Normal cerebrospinal fluid (CSF) leukocytes were seen in 18% of patients, and CSF protein was elevated in 70% of patients. Case fatality was 6.7%. Permanent care in hospital needed by 13% of patients, and after 24 months 88% were ambulatory. Prognosis is quite good in myelitis, and normal CSF leukocytes do not exclude myelitis. PMID- 9415535 TI - The effects of high-dose intravenous methylprednisolone on event-related potentials in patients with multiple sclerosis. AB - The aim of this study was to assess the effects of high-dose (i.e. 1000 mg per day) intravenous methylprednisolone (HDMP) on event-related potentials (ERPs), elicited by a standard auditory 'oddball' paradigm, in patients with clinically active multiple sclerosis. In a double-blind study design, forty-four consecutive inpatients were randomly assigned in two clinically similar groups of 22 subjects each; one treated with HDMP for five days, and other with placebo. ERPs were recorded before and after the treatment. After HDMP therapy the P3 peak latency was significantly shortened (P=0.006), while peak latencies of other waves (i.e. N1, P2, and N2) remained unchanged. On the other hand, ERPs were uninfluenced by placebo treatment. Our results suggest the beneficial effect of intravenous HDMP therapy on, at least some aspects of, cognitive processing capabilities (as assessed by the auditory ERPs) in patients with multiple sclerosis. PMID- 9415536 TI - Inactivation of alpha1-antiproteinase (alpha1-AT) and changes in antioxidants' plasma levels in subarachnoid hemorrhage. AB - Recent studies have suggested that a quantitative or a qualitative imbalance between the activity of proteases and its inhibitors hypothetically might be involved in intracranial aneurysm rupture. In the present study we test the hypothesis that the systemic reduction of alpha1-antitrypsin activity might be related to the elevated oxidative potential exerted by cigarette smoking and/or to a systemic low antioxidant capacity. We studied, in a series of 57 patients bearing intracranial aneurysms, the relationship between alpha1-antitrypsin activity, cigarette smoking and the following variables measured in plasma: vitamin A, vitamin E, thiol groups, urate and lipid peroxide levels. Serum levels of alpha1-antitrypsin are higher in patients with subarachnoid hemorrhage than in cases of unruptured aneurysms, while the levels of vitamin A and vitamin E are significantly lower in patients that suffered subarachnoid hemorrhage than in controls. Both vitamin A and E levels are related to the occurrence of rupture of the aneurysm, as elicited by logistic regression analysis (P=0.017 and P=0.014, respectively), with a protective effect of higher levels of the variables, as shown by their odds ratio (0.028 and 0.84, respectively). No significant changes in the strength of the association could be appreciated when controlling for smoking habit. None of the other tested variables could be related to the occurrence of the aneurysm rupture. Both alpha1-antitrypsin serum level and the level of vitamin A appeared to be independently related to alpha1-antitrypsin collagenase inhibitory capacity percentage (P=0.03 and P=0.025), with no independent influence of the type of aneurysm and the smoking habit. The results of the present study show that the qualitative pattern of alpha1-antitrypsin is significantly related to the serum level of liposoluble vitamin A, while the type of aneurysm and the smoking habit have no independent influence. This suggests that in a situation in which systemic levels of vitamin A are reduced, the risk of a reduced activity of alpha1-antitrypsin as controller of proteases is elevated, with the consequent increased risk of aneurysm bleeding. PMID- 9415537 TI - Mitochondrial DNA defects in Brazilian patients with chronic progressive external ophthalmoplegia. AB - We report herein on eleven Brazilian patients with mitochondrial DNA (mtDNA) deletions, found among thirteen patients with chronic progressive external ophthalmoplegia (CPEO) and ragged-red fibers (RRF). The molecular data was correlated with the morphological and clinical findings. The muscle biopsies were studied by histochemistry, immunohistochemistry and DNA analysis. Muscle mtDNA deletions were mapped and quantitated by Southern blot analysis, polymerase chain reaction and sequencing. Of the eleven patients, ten had CPEO without multisystemic involvement and one had Kearns-Sayre syndrome. Three patients had multiple deletions, two of them with no apparent family history. Eight patients showed heteroplasmic single deletions, ranging in length from 2309 to 7566 bp; three of them had the same 'common deletion' of 4977 bp. The proportion of deleted mtDNA ranged from 14 to 89%. Immunohistochemical studies revealed decreased reactivity with the mtDNA-encoded subunit II of cytochrome c oxidase (COX) in all patients, but preserved activity with the nuclear-encoded COX subunit IV in COX-deficient fibers. Two cases presented a few COX-negative fibers with reduced COX IV immunostaining. We found a high frequency of mtDNA deletions in Brazilian patients with CPEO. There was no correlation between clinical severity, morphological findings and the size or amount of the mutated mtDNA in muscle, suggesting that there are still unknown factors influencing the disease phenotype. PMID- 9415538 TI - CAG repeat length and disease duration in Machado-Joseph disease: a new clinical classification. AB - To evaluate the clinical characteristics of Machado-Joseph disease (MJD) with reference to CAG repeat length and disease duration, we analyzed neurologic findings in 108 patients from 84 families. The majority of MJD patients presented with an ataxic gait as the initial symptom. Dysarthria and nystagmus were observed from an early stage. Bulging eyes, muscle atrophy and bradykinesia developed later. Patients with a shorter CAG repeat length or later onset had more frequent involvement of proprioceptive sensory deficit. Incidence of abnormal reflexes, tones, and proprioceptive sensation was not associated with disease duration, but with CAG repeat length. Based on these results, we propose a new clinical classification: type A (juvenile type), with hyperreflexia and dystonia, but without a proprioceptive sensory deficit; type C (adult type), with hyporeflexia and a proprioceptive sensory deficit, but without dystonia; and type B (intermediate type), the remaining patients with a mixed presentation. PMID- 9415539 TI - Modified total lymphoid irradiation and low dose corticosteroids in progressive multiple sclerosis. AB - In a double-blind prospective randomized trial, we assessed the efficacy and safety of modified total lymphoid irradiation (TLI) plus low dose prednisone (TLI LDP) as compared to sham TLI plus identical prednisone therapy (sham TLI-LDP) in 46 patients with progressive forms of multiple sclerosis (MS). No significant difference existed between groups at study entry in patient age, sex, duration of MS, or disability status. However, following treatment, significantly fewer TLI patients showed a sustained one point decline in the Expanded Disability Status Scale, the primary study endpoint, as compared to the sham TLI group using the Kaplan-Meier Product-limit survival analysis, (P<0.005). Risk for relapse requiring treatment with intravenous methylprednisolone was reduced by 54% in the TLI-treated group (P<0.05). Significantly fewer TLI-LDP patients had gadolinium enhancing plus new T2-weighted lesions (P=0.018) when compared to the sham group post-treatment. There was also a substantial and significant decrease in blood lymphocytes in the TLI-LDP group when compared to either pretreatment values or to sham TLI-LDP through at least 12 months post-therapy. Side effects secondary to TLI were generally mild and well-tolerated. These results further support the hypothesis that TLI and systemic immunosuppression have a beneficial effect in progressive forms of MS. PMID- 9415541 TI - Delayed segmental axial dystonia of the trunk on standing after lumbar disk operation. AB - We report four patients with various degrees of chronic, tonic, mildly painful, or non-painful, kyphoscolioses in orthostatism, which developed weeks, or months, after one or several laminectomies for lumbar disk hernia, in the absence of recurring radicular pain or acute lumbar pain. No family history or personal antecedent, of focal or generalized dystonia was found and the dystonia was not seen in any of the four patients pre-operatively, or during the immediate post operative period. Only ill-defined lumbar 'discomfort', unlike their pre operative lumbago, was reported by the patients, before and during the occurrence of the pathologic trunk posture on standing. Asymmetric lumbar muscle tonic contraction and hypertrophy was found on physical examination. In all patients, the kyphoscoliosis was maximal when standing, partially disappeared when seated, and completely when lying down. One patient responded well to clonazepam, but the other three showed no improvement with either clonazepam or local injections of botulinum toxin; L-dopa was ineffective in all cases, and trihexiphenidyle in three. PMID- 9415542 TI - Aberrant glycosylation/phosphorylation in chromatolytic motoneurons of Werdnig Hoffmann disease. AB - Chromatolytic motor neurons (cMN) in Werdnig-Hoffmann disease (WHD) were investigated in both spinal anterior horns and hypoglossal nuclei with both immuno- and lectin-histochemistry in six cases (3-9 months; two female and four male) of clinically typical WHD. Most characteristic findings from lectin histochemistry were central accumulation of N-linked glycopeptides and marked general paucity of O-linked glycopeptides in cMN. Phosphorylated intermediate filaments, developmentally regulated cytoskeletons and cell adhesion molecules were abundant at the periphery of cMN, as visualized with immunohistochemistry. Both N-linked and O-linked glycoproteins were reciprocally absent or scarce at the peripheral zone of cMN. This intriguing phenomenon provided the basis for postulating the pathogenesis of WHD. The reciprocal ('yen-yang') dissociation of phosphorylation and glycosylation of neurofilament proteins was only seen with phosphorylated neurofilaments and seen only in cMN, not in the control or surrounding unaffected motoneurons. The central accumulation of N-linked glycopeptides was in contrast with peripheral absence of O-GlcNAc-linked glycopeptides which would normally be expected to colocalize with phosphorylated neurofilaments. Both O-glycosylation and phosphorylation are considered essential for assembly and network of neurofilaments. Aberrant O-glycosylation and dissociation of O-glycosylation/phosphorylation would not only cause a defect in neurofilament assembly but also neuron-glia adhesion (via a molecule such as Ng CAM), causing a failure of lower motoneurons to synapse homophilically with the upper motoneurons and also a failure to adhere heterophilically to glia, resulting in the histopathologic tetrad ((i) central chromatolysis, (ii) empty cell beds, (iii) migratory motoneurons and (iv) glial bundles of spinal roots) typical of WHD. PMID- 9415540 TI - The effect of total lymphoid irradiation and low-dose steroids on T lymphocyte populations in multiple sclerosis: correlation with clinical and MRI status. AB - We have monitored the cell surface phenotypic changes occurring in T, B and NK cells of chronic progressive multiple sclerosis (MS) patients after total lymphoid irradiation (TLI) plus low-dose prednisone (TLI-LDP) therapy in comparison to sham TLI-LDP. TLI-LDP resulted in a marked reduction in the relative and absolute number of total CD3+ T cells, CD4+ helper T cells, CD4+ CD45RA+ naive T cells and CD19+ B cells for at least 1 year after treatment. No change occurred in the percent CD8+ T cells although the number of these cells declined after radiotherapy. The CD4/CD8 T cell ratio was also decreased. The relative percent of CD16+ NK cells increased steadily after TLI-LDP while the number of NK cells transiently declined but returned to baseline values 1 year later. An increase in the percent of CD2+ CD3- cells and a decrease in their number after therapy was also observed. In contrast, no significant changes in the number of T, B or NK cells were seen in the MS patients receiving sham TLI LDP. These results provide further evidence that radiotherapy causes a reduction of immunocompetent T and B cells and that a population of possibly NK cells and/or immature T cells appears to be repopulating the circulation after TLI. In addition, a correlation was observed between alterations in lymphocyte populations and the presence or absence of contrast enhancing or new T2 lesions on brain magnetic resonance imaging (MRI) in the TLI-LDP treated MS patients. Patients devoid of contrast enhancing or new T2 lesions had a decreased percentage of CD3+ and CD4+ T cells prior to therapy and at six months following TLI-LDP compared to patients with such lesions. An association was also observed between stability in disease activity as determined on the Expanded Disability Status Scale and relative values of CD3 T cells. PMID- 9415543 TI - Relationship between brainstem MRI and pathological findings in progressive supranuclear palsy--study in autopsy cases. AB - The relationship between the features of MRI in brainstem and pathological findings was investigated in eight autopsy cases with progressive supranuclear palsy (PSP). Features of T1-weighted images at midbrain level were atrophy of tegmentum and tectum, and dilatation of aqueduct. Histologically, these findings were consistent with atrophy of periaqueductal gray matter, quadrigeminal plate, and tegmentum. In these lesions, we detected neuronal loss, decrease in density of myelinated fibers, gliosis, rarefaction of tissues, and tau-positive structures such as neurofibrillary tangles (NFTs), glial fibrillary tangles (GFTs) and neuropil threads. At pons level, atrophy of tegmentum, atrophy of pontine base, and dilatation of prepontine cistern were found. Tau-positive structures were observed not only in tegmentum but also in pontine base. The density of the tau-positive structure was closely related to the severity of atrophy. Features of T2-weighted images were high intensity in the periaqueductal lesion and tegmentum in pons. In these lesions, severe histological findings were detected. The MRI features in brainstem were closely related to the histological findings as PSP. PMID- 9415544 TI - Cryoglobulinaemic neuropathy manifesting with restless legs syndrome. AB - In a series of 12 patients with essential mixed cryoglobulinaemia (EMC) and peripheral neuropathy as main feature of the disease, restless legs syndrome (RLS) was a major manifestation in four women, aged 55-65 years. In one patient RLS was a presenting manifestation of the disease, and in another patient the diagnosis of EMC was made investigating RLS and polyneuropathy, although prior rheumatological symptoms were retrospectively recognized. All patients with RLS had symmetrical sensory polyneuropathy, but non-RLS patients had also other forms of peripheral neuropathy, and symmetrical sensory polyneuropathy only in two of eight cases (P=0.03). Neurophysiological study showed that sensory action potentials of the sural nerve were more often inelicitable in non-RLS patients (six of eight) than in RLS patients (none of three). Sural nerve biopsy had no distinctive features in three RLS patients, with regard to other patients with cryoglobulinaemic neuropathy. RLS seems not uncommon in cryoglobulinaemic neuropathy, and significantly associated with symmetrical sensory polyneuropathy, whereas patients with other subtypes of cryoglobulinaemic neuropathy do not develop RLS; thus, a disorder of the sensory inputs may be important in the pathogenesis of RLS. The occurrence of RLS, especially in middle-aged women, should prompt investigations for peripheral neuropathy focusing on cryoglobulinaemic neuropathy. PMID- 9415545 TI - Synthetic growth hormone has no inducting effect in the development of a prion disease: an experimental study on the scrapie model in hamsters. PMID- 9415547 TI - Pick's disease with amyotrophic lateral sclerosis. PMID- 9415546 TI - Superoxide dismutase activity in amyotrophic lateral sclerosis. PMID- 9415548 TI - Molecular HLA typing--the brave new world. PMID- 9415549 TI - Prostaglandin E1 protects against ischemia-reperfusion injury of the liver by inhibition of neutrophil adherence to endothelial cells. AB - BACKGROUND: This study investigates the protective mechanism of prostaglandin E1 (PGE1) against hepatic ischemia-reperfusion injury in vivo. It has been demonstrated that activated leukocytes contribute to ischemia-reperfusion injury, and that administration of the monoclonal antibody (mAb) for adhesion molecules reduces the injury by inhibiting leukocyte-endothelial cell adhesion. We therefore attempted to find out whether PGE1 has an effect on the inhibition of neutrophil adherence to endothelial cells after reperfusion. METHODS: We administered anti-intercellular adhesion molecule 1 (ICAM-1) mAb, antiserum against rat polymorphonuclear leukocytes, or PGE1 to a rat model of left lobar ischemia for 60 min followed by reperfusion. Leukocyte adherence was observed by intravital fluorescence microscopy. The effect of PGE1 on the expression of adhesion molecules was analyzed by immunohistochemistry and flow cytometry. RESULTS: Ischemia-reperfusion caused endothelial dysfunction and hepatocellular injury with leukostasis in postsinusoidal venules. Anti-ICAM-1 mAb administration or leukopenia ameliorated both the hepatocellular injury and endothelial dysfunction. Although PGE1 administration did not affect the serum interleukin-8 level, it significantly decreased hepatic injury and leukostasis in the reperfused liver. Immunohistochemical findings showed that PGE1 decreased ICAM-1 expression on endothelial cells, but did not affect lymphocyte function associated antigen 1, and membrane attack complex 1 on neutrophils in flow cytometric analysis. CONCLUSIONS: We conclude that PGE1 protects the liver against ischemia-reperfusion injury by reducing leukocyte-endothelial cell adhesion via down-modulation of ICAM-1 expression on the endothelium. PMID- 9415551 TI - Combination therapy with cyclosporine and interleukin-4 or interleukin-10 prolongs survival of synergeneic pancreatic islet grafts in nonobese diabetic mice: islet graft survival does not correlate with mRNA levels of type 1 or type 2 cytokines, or transforming growth factor-beta in the islet grafts. AB - BACKGROUND: The recurrent autoimmune response to syngeneic pancreatic islet grafts transplanted into nonobese diabetic (NOD) mice is cell-mediated and relatively resistant to cyclosporine (CsA) therapy. Therefore, we asked whether interleukin (IL)-4 and IL-10, cytokines that inhibit cell-mediated immunity, might improve the therapeutic effect of CsA. METHODS: We compared the survival of syngeneic islet grafts transplanted into diabetic NOD mice treated with IL-4, IL 10, and CsA, administered as single agents and in combinations. Additionally, we measured mRNA levels of type 1 cytokines (interferon-gamma [IFN-gamma], IL-2, and IL-12), type 2 cytokines (IL-4 and IL-10), and transforming growth factor-beta (TGF-beta) to determine whether graft rejection or survival might correlate with expression of these cytokines in the grafts. RESULTS: CsA (20 mg/kg/day) significantly prolonged islet graft survival (median: 20 days vs. 10 days for vehicle-treated mice). Neither IL-4 (2.5 microg, twice daily), nor IL-10 (10 microg, twice daily) significantly prolonged islet graft survival. By contrast, combination therapy with CsA and IL-10 significantly prolonged islet graft survival (median: 34 days) compared with vehicle-treated mice (median: 10 days), and combination therapy with CsA and IL-4 significantly prolonged islet graft survival (median: 59 days) compared with both vehicle-treated mice (median: 10 days) and mice treated with CsA alone (median: 20 days). Islet grafts from normoglycemic mice treated with CsA plus IL-10, and with CsA plus IL-4, were surrounded but not infiltrated by mononuclear leukocytes and beta cells were intact, whereas islet grafts from mice treated with vehicle, IL-4, IL-10, and CsA (as single agents) were infiltrated by mononuclear leukocytes and fewer beta cells were detected. Polymerase chain reaction analysis of cytokine mRNA expression in islet grafts at 8-12 days after transplantation revealed that CsA decreased mRNA levels of type 1 cytokines (IFN-gamma and IL-12p40), whereas CsA plus IL-10 did not, and CsA plus IL-4 increased mRNA levels of IFN-gamma, IL 12p40, and TGF-beta. CONCLUSIONS: These results demonstrate that IL-4, and to a lesser extent IL-10, improves the ability of CsA to prevent autoimmune destruction of beta cells in syngeneic islets transplanted into diabetic NOD mice; however, there is no simple correlation between the protective effects of the different treatment regimens (CsA, CsA plus IL-4, and CsA plus IL-10) and mRNA levels of type 1 cytokines (IFN-gamma, IL-2, and IL-12), type 2 cytokines (IL-4 and IL-10), or TGF-beta in the islet grafts. PMID- 9415550 TI - Prevention of late renal changes after initial ischemia/reperfusion injury by blocking early selectin binding. AB - BACKGROUND: Increasing clinical evidence suggests that delayed initial function secondary to ischemia/reperfusion injury alone, and particularly in combination with early episodes of acute rejection, reduces kidney allograft survival over time. METHODS: We investigated changes developing over the long term following a standardized ischemia/reperfusion insult in a Lewis rat model. The left kidney was isolated in a uninephrectomized host and cooled, and the pedicle was clamped for 45 min. Animals were followed for 48 weeks after initial renal injury. Organs were removed serially (4, 8, 16, 24, 32, 40, and 48 weeks) for immunohistology and reverse transcriptase polymerase chain reaction. RESULTS: Progressive proteinuria developed after 8 weeks. By immunohistology, CD4+ leukocytes and ED 1+ macrophages infiltrated the ischemic organs in parallel with up-regulation of major histocompatibility complex class II antigen expression. Because macrophages have been shown to be critical in chronic changes in other models, they were examined primarily in these studies. By reverse transcriptase polymerase chain reaction, macrophage-derived, fibrosis-inducing factors (transforming growth factor-beta, interleukin 6, and tumor necrosis factor-alpha) remained highly and constantly expressed throughout the follow-up period. The long-term influence of initial treatment with the soluble form of P-selectin glycoprotein ligand-1, a soluble ligand for P- and E-selectin, was then examined. All functional and structural changes remained at relative baseline, similar to uninephrectomized controls. CONCLUSIONS: These data suggest that blocking the initial selectin mediated step after ischemia/reperfusion injury, which triggers significant early cellular and molecular events, also reduces later renal dysfunction and tissue damage over time. In part, the findings may be explained by the sparing of functioning nephron units, which if destroyed or compromised by the original insult, may contribute to long-term graft failure. This approach may be important clinically in the transplantation of kidneys from non-heart-beating or marginal donors or organs experiencing prolonged ischemic times. PMID- 9415552 TI - Alloantibody- and T cell-mediated immunity in the pathogenesis of transplant arteriosclerosis: lack of progression to sclerotic lesions in B cell-deficient mice. AB - BACKGROUND: The relative roles of humoral and cell-mediated immunity in generating chronic allograft arteriopathy have been considered for several years. We have sought definitive evidence regarding these questions using heart transplants between mouse strains selected to isolate the effects of each form of immune responsiveness. METHODS: B10.BR hearts were transplanted to B cell deficient recipients that are devoid of immunoglobulins (muMT). Their vessels were compared with those of transplants to fully reactive recipients of the same genetic background (C57BL/6). Additional evidence came from comparisons in other strain combinations. RESULTS: Transplants to B cell-deficient and normal recipients developed cellular coronary endothelialitis, with destruction of the arterial media, accompanied by the adherence of T lymphocytes and macrophages to endothelial surfaces. In B cell-deficient recipients, there was no centripetal migration of smooth muscle, alpha-actin-positive myointimal cells and little deposition of collagen or ground substance, compared with lesions in fully reactive C57BL/6 recipients in which these changes are prominent. In two other donor-recipient combinations in which anti-donor antibodies are generally undetectable (B10.BR-->B10.A and 129-->C57BL/6), intimal fibrosis was uncommon. However, B10.A recipients became capable of producing fibrous lesions in B10.BR hearts when given anti-donor, class I antibody by passive transfer, as we have observed previously in scid recipients. CONCLUSIONS: Taken together, these findings indicate that endothelialitis is antibody-independent, whereas antibodies potentiate and can be sufficient for fully developed, fibrous, chronic allograft vasculopathy. Therapeutic strategies for controlling chronic lesions must consider inhibition of the humoral response. PMID- 9415553 TI - In situ gene transfer into rat auxiliary liver transplant. AB - BACKGROUND: A replication-defective retrovirus BAG vector was tested for in situ delivery of the beta-galactosidase gene to auxiliary liver transplant in a rat model. METHODS: The BAG vector, which was shown to be effective in genetic transduction of cultured NIH/3T3 cells, was produced in a psi2 packaging cell and later amplified in a selected PA317 clone. Hepatocyte replication was induced by one-third hepatectomy of the donor liver, and the procedure was followed by auxiliary partial liver transplantation. Twenty-four hours after hepatic induction or transplantation, viral supernatant at 37 degrees C was perfused into the liver graft via the portal vein during a temporary occlusion of the graft portal vein. RESULTS: All animals survived the transplantation procedures and were killed at specified time intervals. Histochemical staining of the liver graft specimens indicated the expression of beta-galactosidase in the gene transferred group but not in the control animals. As demonstrated by polymerase chain reaction assay, the proviral beta-galactosidase sequence was present in the graft specimens, but absent from all other tissues tested. CONCLUSIONS: In short, the retrovirus BAG vector can be useful for in situ delivery of foreign genes to liver graft in transplantation and other clinical settings, providing a simple, consistent, and reliable alternative in hepatic gene therapy experiments. PMID- 9415554 TI - Engraftment of human kidney tissue in rat radiation chimera: I. A new model of human kidney allograft rejection. AB - BACKGROUND: We have recently shown that lethally irradiated normal strains of mice and rats, reconstituted with bone marrow from severe combined immune deficiency (SCID) mice, can be engrafted with human peripheral blood mononuclear cells (PBMC). METHODS: The feasibility of transplanting human renal tissue under the kidney capsule of the SCID/Lewis and SCID/nude radiation chimera and the effects of intraperitoneal infusion of allogeneic human PBMC on the human renal implants were investigated by histology, electron microscopy, immunohistochemistry, and fluorescence-activated cell sorter analysis. RESULTS: Sequential evaluation of the human renal implants from 10 days to 2 months after transplantation showed that human parenchymal elements survive in the implants up to 2 months after transplantation. The overall architecture of the transplanted kidney tissue and the normal structure of individual cells in the glomeruli and tubuli were preserved. Infusion of allogeneic human PBMC after kidney implantation resulted in patchy cellular infiltrates, composed mainly of activated human T cells, and led to prompt rejection of the human renal tissue, whereas no signs of inflammation were observed in human renal implants of chimeric rats that did not receive human PBMC. Treatment with OKT3 antibody, anti human CD25 antibody, or CTLA4Ig fusion protein in vivo ameliorated the rejection process. CONCLUSIONS: Human adult kidney fragments transplanted into SCID-like rats transiently retain competent parenchymal structures. When these grafts are combined with allogeneic human PBMC, acute cellular rejection develops. We suggest that this chimeric model might be useful for the investigation of the effects of experimental manipulation on the kinetics of the inflammatory response during human renal allograft rejection. PMID- 9415555 TI - Engraftment of human kidney tissue in rat radiation chimera: II. Human fetal kidneys display reduced immunogenicity to adoptively transferred human peripheral blood mononuclear cells and exhibit rapid growth and development. AB - BACKGROUND: Transplantation of human kidney tissue under the kidney capsule of immunodeficient animals (severe combined immunodeficiency [SCID]/Lewis and SCID/nude chimeric rats), and the subsequent intraperitoneal infusion of allogeneic human peripheral blood mononuclear cells (PBMC), results in a rapid and consistent human renal allograft rejection. We investigated the consequences of grafting human fetal kidney fragments instead of the adult tissue. METHODS: The development of human fetal kidney tissue and its interaction with allogeneic human PBMC in chimeric rats were analyzed by histology, immunohistochemistry, and in situ hybridization. RESULTS: We report successful establishment of human fetal kidney to SCID/Lewis and SCID/nude chimeric rats. The intrarenal human fetal renal implants displayed rapid growth and maintained numerous developing glomeruli and tubular structures up to 4 months after transplantation. In contrast to the adult human kidney, infusion of allogeneic human PBMC resulted in either minimal human T-cell infiltration or abundant nonrejecting T-cell infiltrates, characterized by a reduced number of T cells of the CD45RO+ or HLA DR+ subsets, both leading to less tissue destruction as well as to continued growth of the human fetal renal tissue. This observation was found to be related to the reduced protein expression of tissue HLA class I and II, intercellular adhesion molecule 1, and vascular adhesion molecule 1 in the fetal grafts compared with the adult grafts. Lack of tissue expression of Fas ligand in the fetal grafts suggests that the latter does not contribute to the delayed rejection of human fetal kidneys. CONCLUSIONS: Our model should be useful for the study of human fetal renal development and the human alloresponse against fetal tissue. PMID- 9415556 TI - Induction of tolerance with nondepleting anti-CD4 monoclonal antibodies is associated with down-regulation of TH2 cytokines. AB - BACKGROUND: Induction of tolerance with anti-CD4 has mainly focused on monoclonal antibodies (mAbs) that deplete CD4+ T cells. In this study, the mechanisms by which nondepleting anti-CD4 mAbs induce tolerance in the Dark Agouti to PVG rat heart graft model were examined. METHODS: Five anti-CD4 mAbs were tested. Immunohistology and cytokine mRNA profiles were analyzed within grafts. Effects of combining anti-CD4 therapy with alloantibody (alloAb), interleukin (IL)-4, and anti-IL-4 mAb were also examined. RESULTS: All mAbs tested induced indefinite graft survival (>150 days), with blocking of alloAb production. Exogenous alloAb did not restore rejection. Similar T cell receptor alphabeta+, CD8+, IL-2 receptor+ T cell, macrophage, and natural killer cell infiltration and comparable MHC II and intercellular adhesion molecule-1 levels were seen in rejecting and tolerant grafts. mRNA for IL-2, interferon-gamma, lymphotoxin, tumor necrosis factor-alpha, transforming growth factor-beta, cytolysin, and granzyme-A/B was comparable, although inducible nitric oxide synthase was slightly reduced in tolerant grafts. IL-4 and IL-5 were significantly reduced in tolerant grafts, although IL-6, IL-10, and IL-13 levels were similar; this was consistent with partial T helper (Th)2 response inhibition, which was also manifested by inhibited alloAb. The combination of alloAb, IL-4, or anti-IL-4 mAb with anti-CD4 did not prevent tolerance induction. CONCLUSIONS: This study demonstrated that anti-CD4 mAb therapy did not inhibit activation and infiltration of Th1 and CD8+ effector T cells. Preferential induction of Th2 responses, especially IL-4, was not essential for the induction of tolerance. Our studies also found no evidence to support induction of anergy or transforming growth factor-beta as mechanisms of tolerance induction. These results question whether IL-4 is required for induction of transplantation tolerance. PMID- 9415557 TI - Percutaneous management of a bladder-drained pancreas transplant pseudocyst by a transcystic approach. AB - BACKGROUND: We describe a 35-year-old male type 1 diabetic who underwent a cadaveric combined kidney-bladder-drained pancreas transplant with a duodenocystostomy for exocrine drainage who developed a large pelvic pseudocyst associated with a dilated pancreatic duct and an elevated serum amylase level. METHODS: Due to the risk of surgical revision and the possibility of creating a cutaneous fistula with conventional percutaneous drainage, a pseudocyst-to bladder drainage was performed. After the procedure, the catheter was capped to allow drainage of the pancreatic secretions into the bladder. RESULTS: After drainage, the patient's serum amylase and lipase normalized along with resolution of the pseudocyst. The tube was removed after 19 weeks with no evidence of a recurrent pseudocyst and a normal serum amylase level. CONCLUSION: The percutaneous pseudocyst-cystostomy obviated the need for surgical revision of the exocrine gland drainage and thus eliminated the morbidity and the potential risk of graft loss associated with such surgery. PMID- 9415558 TI - Solitary pancreas transplantation for nonuremic patients with labile insulin dependent diabetes mellitus. AB - BACKGROUND: Simultaneous pancreas-kidney transplantation has become a widely accepted treatment option for selected uremic patients with insulin-dependent diabetes mellitus (IDDM). Patient survival rates at 1 year exceed 90%, and rates of pancreas graft survival, 70%. However, solitary pancreas transplantation for nonuremic patients with IDDM has been controversial because of the less favorable outcome and the need for long-term immunosuppression with its associated morbidity and mortality. METHODS: We studied the outcome of 225 solitary pancreas transplants during three immunosuppressive eras: the precyclosporine (CsA) era (n=83), the CsA era (n=118), and the tacrolimus era (n=24). Only patients with labile IDDM (e.g., hypoglycemic unawareness, insulin reactions, > or = 2 failed attempts at intensified insulin therapy for metabolic control) underwent solitary pancreas transplantation. Using univariate and multivariate analyses, we looked at patient and graft survival, the risk of surgical complications, and native kidney function during these three eras. RESULTS: Pancreas graft survival improved significantly over time: 34% at 1 year after transplantation in the pre CsA era, 52% in the CsA era, and 80% in the tacrolimus era (P=0.002). Pancreas graft loss due to rejection decreased from 50% at 1 year in the pre-CsA era, to 34% in the CsA era, to 9% in the tacrolimus era (P=0.008). The rate of technical failures (i.e., the risk of surgical complications) decreased from 30% in the pre CsA era, to 14% in the CsA era, to 0% in the tacrolimus era (P=0.001). Patient survival rates at 1 year have ranged between 88% and 95% in the three eras (P=NS). Matching for at least one antigen on each HLA locus and avoiding HLA-B mismatches significantly decreased the incidence of rejection. The incidence of native kidney failure due to drug-induced toxicity decreased significantly over time, in part because only recipients with pretransplant creatinine clearance > or = 80 ml/min received transplants. CONCLUSIONS: Solitary pancreas transplantation has become a viable alternative for nonuremic patients with labile IDDM. The risks of surgical complications and drug-induced nephrotoxicity have significantly decreased over time. Using tacrolimus as the mainstay immunosuppressant, patient and graft survival rates now no longer trail those of simultaneous pancreas-kidney transplantation. PMID- 9415559 TI - Long-term allograft acceptance in a patient with posttransplant lymphoproliferative disorder: correlation with intragraft viral interleukin-10. AB - BACKGROUND: Viral (v) interleukin (IL)-10 is expressed by Epstein-Barr virus (EBV) and has pro- and anti-inflammatory actions similar to human IL-10. EBV is also a known factor in the development of posttransplant lymphoproliferative disorder (PTLPD) in allograft recipients. We observed a patient with widespread PTLPD 9 months after renal transplantation, who subsequently maintained renal function despite minimal immunosuppression, and we investigated a possible link between these factors. METHODS: The patient's chart was reviewed for relevant history. EBV DNA in blood and tissues was assessed by polymerase chain reaction. Human and vIL-10 and gamma-interferon mRNA were evaluated with reverse transcription-polymerase chain reaction using nested primers. RESULTS: After the diagnosis of PTLPD, the patient was maintained on prednisone (8 mg/day) as the only immunosuppression with preserved renal function for 17 months until death as a result of pulmonary failure. She had continuously high blood levels of EBV DNA, although only mild persistent intrarenal atypical lymphocytic infiltrates. Human IL-10 mRNA was never present; in contrast, intragraft vIL-10 mRNA was identified and associated with resolution of an intervening episode of severe acute transplant rejection. CONCLUSIONS: We suggest that the preserved renal function resulted from the anti-inflammatory actions of vIL-10 inhibiting acute rejection in the renal allograft. PMID- 9415560 TI - Infectivity of hepatic allografts with antibodies to hepatitis B virus. AB - BACKGROUND: Since suitable recipients for hepatic allografts from donors with antibodies to hepatitis B virus (HBV) have not been determined, a review of our 7 year experience with donors positive for hepatitis B surface antibody (anti-HBs), hepatitis B core antibody (anti-HBc), or both was undertaken. METHODS: Recipients of hepatic allografts from donors with antibodies to HBV were identified by a retrospective review of procurement records and screened for HBV infection. RESULTS: From January 1, 1990, to January 1, 1997, 2578 liver transplants were performed and 140 (5.4%) recipients received an allograft from a donor with antibodies to HBV. Twenty-five of 48 recipients of a hepatic allograft from a donor positive only for anti-HBs were screened and none developed HBV infection. Twenty-five of 41 naive recipients of a hepatic allograft from an anti-HBc positive donor were screened and 18/25 (72%) developed HBV infection. Four of these 18 naive recipients with HBV infection received an allograft from a donor positive for both anti-HBc and anti-HBs. Seven of 13 anti-HBs-positive recipients of an allograft from an anti-HBc-positive donor were screened and none developed HBV infection. Fifteen of 16 recipients positive only for anti-HBc who received a hepatic allograft from an anti-HBc-positive donor were screened and 2/15 (13%) developed HBV infection. CONCLUSIONS: Hepatic allografts from donors positive only for anti-HBs do not transmit HBV infection. Hepatic allografts from anti-HBc positive donors frequently transmit HBV infection to naive recipients regardless of the donor anti-HBs status, and antiviral prophylaxis may be indicated. Anti HBs-positive recipients appear resistant to HBV infection after orthotopic liver transplantation with an allograft from an anti-HBc-positive donor. Recipients positive only for anti-HBc infrequently develop HBV infection when transplanted with an allograft from an anti-HBc-positive donor; however, HBV prophylaxis may be justified. PMID- 9415561 TI - Cholic acid synthesis is reduced in pediatric liver recipients during graft dysfunction due to ischemic injury and allograft rejection. AB - BACKGROUND: Bile acids are synthesized and secreted by the liver. During liver failure and hepatic dysfunction, a marked reduction of bile acid synthesis has been shown. The purpose of this study was to determine whether the biliary bile acid pattern was affected by preservation injury and rejection and whether it was a reliable marker for graft function in pediatric liver recipients after liver transplantation. METHODS: We prospectively measured the biliary bile acid pattern in 126 serial bile samples obtained from 15 consecutive pediatric liver recipients by reversed phase high pressure liquid chromatography and correlated our results with clinical findings: preservation injury, no rejection, rejection, or infection. RESULTS: There was a significant change of the bile acid pattern during the first 3 days after transplant. Total biliary bile acids, cholic acid (CA), and CA/chenodeoxycholic acid (CDCA) ratio increased in 12 of 15 patients with mild preservation injury. These changes of the bile acid pattern were markedly delayed in patients with severe preservation injury. During 16 rejection episodes, total biliary bile acid, CA, and CA/CDCA ratio decreased significantly, but returned to normal after successful treatment of rejection. Bacterial infection, observed in nine children, and cyclosporine toxicity, observed in three children, seemed to have no affect on the biliary bile acids. CONCLUSIONS: Liver cell damage as a result of preservation injury or rejection leads to a reduction of biliary CA, resulting in a decrease of total biliary bile acids and the CA/CDCA ratio in pediatric liver recipients. This might be caused by a diminished secretion of bile acids and by a decreased synthesis of bile acids. PMID- 9415562 TI - High prevalence of anemia after cardiac transplantation in children. AB - BACKGROUND: Chronic anemia is common in adults after successful cardiac transplantation. However, the prevalence of anemia in children after cardiac transplantation is uncertain. The purpose of this study was to investigate the prevalence and causes of chronic anemia in well children after cardiac transplantation and in particular to define the role, if any, of iron deficiency, which is important and relatively common in normal children. METHODS: Twenty children (ages 7 months to 16 years) who were well 4 months to 6 years after cardiac transplantation were studied. Fourteen children (70%) were anemic and enrolled in a prospective trial of iron supplementation. RESULTS: In the majority of children, serum iron and erythropoietin levels were low, although serum ferritin and zinc protoporphyrin levels tended to be normal or high. Only one child demonstrated a definite response to iron supplementation, although the hemoglobin level remained low. CONCLUSIONS: Anemia is highly prevalent in this population, and, despite the presence of low serum iron and transferrin saturation, anemia is not usually due to iron deficiency. Although the diagnosis of iron deficiency in this group is difficult and must not be missed, inappropriate therapy should be avoided. In the majority of children, there appears to be an anemia of chronic disease which may be secondary to chronic inflammation or an effect of cyclosporine on erythropoietin production. PMID- 9415563 TI - Evidence for engraftment of human bone marrow cells in non-lethally irradiated baboons. AB - BACKGROUND: Prior to organ harvesting, an attempt was made to modulate the donor's immune responses against prospective xenogeneic recipients by infusion of "recipient-type" bone marrow. METHODS: For this purpose, baboons conditioned with total lymphoid irradiation were given 6 x 10(8) unmodified human bone marrow cells/kg body weight with no subsequent treatment. RESULTS: Animals survived until they were euthanized at 18 months. Using primers specific for human chorionic gonadotrophin gene, the presence of human DNA was confirmed by polymerase chain reaction in the blood of one animal for up to 18 months after cell transplantation; in the other animal, xenogeneic chimerism became undetectable in the blood at 6 months after bone marrow infusion. However, tissue samples obtained from both animals at the time they were euthanized had evidence of donor (human) DNA. Additionally, the presence of donor DNA in individually harvested colonies of erythroid and myeloid lineages suggested that infused human bone marrow cells had engrafted across the xenogeneic barrier in both baboons. CONCLUSIONS: Bone marrow transplantation from human to baboon leads to establishment of chimerism and modulation of donor-specific immune reactivity, which suggests that this strategy could be reproducibly employed to create "surrogate" tolerogenesis in prospective donors for subsequent organ transplantation across xenogeneic barriers. PMID- 9415564 TI - Improved technique of heterotopic cervical heart transplantation in mice. AB - BACKGROUND: The method for mouse vascularized heart transplantation have been described using suture and cuff techniques. Technical problems have limited its widespread use. Here, we describe our method of modified cervical heterotopic transplantation with the cuff technique. METHODS: By using a smaller Teflon cuff (external diameter 0.6 mm, internal diameter 0.4 mm) and superfine-tip forceps, it became possible to directly pull the edge of the carotid artery and evert the proximal end of the artery over the cuff. Similarly, the external jugular vein could be easily everted over a 22-gauge cuff with this direct pulling method. RESULTS: By these modifications, the operation time was reduced. It usually takes 20 min for the donor harvest, 15 min for preparation of the cervical vessels, and 15 min for anastomosis. All procedures from the donor harvest through skin closure of the recipient mice can be completed within 1 hr, and ischemic time is within 25-40 min. CONCLUSIONS: This method can be used to investigate cyclophosphamide-induced tolerance and mechanisms of reperfusion injury. PMID- 9415565 TI - Infection-associated macrophage activation accelerates chronic renal allograft rejection in rats. AB - BACKGROUND: The influence of infection-associated cellular activation on chronic rejection of kidney grafts was assessed in an established rat model by administration of lipopolysaccharide (LPS), an endotoxin and a potent stimulator of various cell populations including mononuclear cells and renal epithelial cells. METHODS: Lewis recipients of F344 kidneys were treated with low-dose cyclosporine (1.5 mg/kg/day x 10 days). Animals with well-functioning grafts received a single dose of LPS (2 mg in 1 ml of NaCl, intraperitoneally) at 4 or 8 weeks after engraftment. Untreated control rats, which later experienced chronic rejection, were given 1 ml of NaCl. RESULTS: Administration of LPS during the early quiescent phase of chronic rejection accelerated the chronic process, functionally (proteinuria), morphologically, immunohistologically, and by reverse transcriptase polymerase chain reaction as compared with untreated controls. Infiltration of macrophages and their associated factors was especially affected. CONCLUSIONS: As the later events of chronic rejection seem to be mediated primarily by macrophages and their products, administration of LPS accelerated the tempo and activity of these cells in the development of chronic rejection. These findings may explain the clinical observation that infection may be an important risk factor for chronic allograft rejection. PMID- 9415566 TI - Living-related intestinal transplantation: first report of a standardized surgical technique. AB - BACKGROUND: Intestinal transplants using cadaver donors have become an alternative to total parenteral nutrition (TPN) for the treatment of irreversible intestinal failure. Intestinal transplants using living-related donors have rarely been attempted, and the surgical technique has not been standardized. METHODS: We performed a living-related intestinal transplant for a paraplegic, 16 year-old boy with life-threatening TPN complications, including lack of vascular access, recurrent line infections, and intermittent liver dysfunction. RESULTS: A four antigen-matched donor (father) underwent resection of 200 cm of the ileum on a vascular pedicle comprising the ileocolic artery and vein. This resection left the donor with 300 cm of proximal small bowel, 20 cm of the most distal terminal ileum, the ileocecal valve, and all of the large intestine. The donor's ileocolic artery and vein were anastomosed to the recipient's infrarenal aorta and cava; bowel continuity was restored with an end-to-end anastomosis between the recipient's jejunum and the donor's ileum. Both donor and recipient had uneventful postoperative courses. Recipient maintenance immunosuppression has been with tacrolimus, mycophenolate mofetil, and prednisone. One year after transplant, urine methylmalonic acid indicates good vitamin B12 absorption in both the donor and recipient. The recipient has been completely off TPN since discharge (posttransplant day 21), has gained 20 kg, and has had no evidence of rejection, infection, or graft-versus-host disease. CONCLUSIONS: Intestinal transplants from living-related donors can be lifesaving for selected patients with chronic intestinal failure and can be done with minimal risk to the donor. PMID- 9415567 TI - Acute respiratory failure and pulmonary fibrosis secondary to administration of mycophenolate mofetil. AB - BACKGROUND: We report the first documented case of pulmonary toxicity to mycophenolate mofetil in this article. METHODS: A 51-year-old woman experienced systemic reactions beginning 10 days after cadaveric renal transplantation. RESULTS: Recurrent respiratory failure and documented progressive pulmonary fibrosis ensued. Cultures were negative and other agents were discontinued. It was not until the mycophenolate was stopped did the patient improve. CONCLUSIONS: Mycophenolate mofetil can cause acute respiratory failure simulating opportunistic infection or pulmonary edema. If not recognized, this may lead to the rapid development of severe pulmonary fibrosis, some of which may not be reversible. PMID- 9415568 TI - Impairment of fibrinolytic potential in long-term steroid treatment after heart transplantation. AB - Thrombotic complications constitute an important risk in transplant recipients, in whom a hypercoagulable state and hypofibrinolysis have been associated with immunosuppressive treatment, especially with cyclosporine. In no case have clotting and fibrinolytic abnormalities been correlated with steroid immunosuppression, even though steroids were always administered. Previous studies found a relationship between hypercorticism and hypofibrinolysis both in Cushing's disease and after renal transplantation. The aim of this investigation was to compare fibrinolytic potential using the venous occlusion test in two similar groups of heart transplant patients treated with or without steroids. Euglobulin lysis time, tissue-type plasminogen activator and plasminogen activator inhibitor-1 (PAI-1) activities, and antigens were determined before and after the venous occlusion test. A reduced fibrinolytic potential (significant prolongation of lysis time) due to a significant increase in PAI-1 activity and antigen levels was found in heart transplant patients treated with steroids, as compared with patients without steroid treatment and control subjects. The prevalence of reduced fibrinolytic potential was 69.2% (18 cases) in the steroid treated group and 34.8% (8 cases) in the non-steroid-treated group. In every case, the impaired fibrinolytic potential was due to high basal PAI-1 levels. Our results are compatible with the presence of a hypofibrinolytic state secondary to long-term steroid treatment. In heart transplant recipients, steroid-induced hypofibrinolysis may constitute a further risk factor for thrombotic disease. PMID- 9415570 TI - HLA antibody screening: comparison of a solid phase enzyme-linked immunoassay with antiglobulin-augmented lymphocytotoxicity. AB - BACKGROUND: IgG antibodies to HLA class I antigens can cause hyperacute rejection of renal allografts. Screening of sera from such transplant candidates is laborious, time-consuming, and expensive when performed by sensitive antihuman globulin-augmented lymphocytotoxicity (AHG-CDC). METHODS: Because 60-70% of our transplant screens are negative, we evaluated a solid phase enzyme-linked method (EIA) as a potential prescreen by parallel testing 215 sera by AHG-CDC and by EIA. This EIA method is designed to detect only IgG antibodies, and all positive AHG-CDC sera were retested after dithiothreitol treatment. RESULTS: There was 96.2% concordance between the tests for IgG antibodies. Seven sera (3.25%) were positive by EIA alone, and one (0.46%) was negative by EIA alone. The EIA method was also less costly ($15.00 versus $105.00) and less time consuming (hours versus days) than AHG-CDC panel testing for large numbers of sera. CONCLUSIONS: We conclude that this EIA method is simple, sensitive, objective, and cost effective as a prescreen for HLA class I antibodies. PMID- 9415569 TI - Long-term ganciclovir prophylaxis for successful prevention of primary cytomegalovirus (CMV) disease in CMV-seronegative liver transplant recipients with CMV-seropositive donors. AB - BACKGROUND: We conducted a trial of long-term ganciclovir prophylaxis for prevention of primary cytomegalovirus (CMV) disease in CMV-seronegative liver transplant recipients with CMV-seropositive donors. METHODS: Patients received intravenous ganciclovir at a dose of 6 mg/kg once a day from day 1 to day 30 after transplant, and then at a dose of 6 mg/kg once a day, Monday through Friday, until day 100. Forty-seven consecutive patients were evaluated. Due to the primary physician's decision or administrative error, 10 patients received less than 7 weeks of ganciclovir (mean duration, 3 weeks). RESULTS: Four of the 10 (40%) patients who received less than 7 weeks of ganciclovir developed CMV disease (hepatitis). In contrast, none of the 37 patients given 100 days of prophylactic ganciclovir developed CMV disease while receiving ganciclovir. Two patients (5.4%) subsequently developed CMV disease (hepatitis) 21 and 88 days, respectively, after completing their ganciclovir prophylaxis. Reversible neutropenia in three patients (8.1%) was the only side effect associated with long-term ganciclovir. Complications from central intravenous catheters did not occur. CONCLUSIONS: These results reaffirm the efficacy and safety of long-term ganciclovir prophylaxis for prevention of primary CMV disease in a large number of high-risk CMV-seronegative liver transplant recipients with CMV-seropositive donors. PMID- 9415571 TI - Administration of OKT3 as a two-hour infusion attenuates first-dose side effects. AB - BACKGROUND: Use of the murine CD3 monoclonal antibody OKT3 is limited by first dose side effects, which are thought to be caused by the release of inflammatory mediators. Because these processes might be influenced by the speed of administration, we compared a 2-hr OKT3 infusion with the bolus infusion usually applied nowadays. METHODS: Eighteen renal allograft recipients were prophylactically treated with OKT3 and randomized to receive the first dose either as a 2-hr infusion or as an intravenous bolus infusion. Clinical side effects score and the occurrence of complement activation, cytokine release, and activation of neutrophils were determined. RESULTS: Two-hour infusion of OKT3 completely prevented the occurrence of dyspnea, reduced the incidence of other side effects, and attenuated complement activation. Cytokine release and depletion of peripheral blood lymphocytes were similar in both groups. CONCLUSIONS: Thus, complement activation seems to play an additional role in the development of side effects after the first OKT3 dose. PMID- 9415572 TI - Efficacy and safety of lamivudine on replication of recurrent hepatitis B after cadaveric renal transplantation. AB - BACKGROUND: The aim of this pilot study was to evaluate the efficacy and the safety of lamivudine therapy in hepatitis B virus (HBV)-positive/DNA-positive renal transplant recipients. METHODS: Six HBV DNA-positive cadaveric renal transplant recipients ranging in age from 49+/-6 years were administered lamivudine, at 100 mg/day for a period of at least 6 months, on a compassionate use basis. Lamivudine is the (-) enantiomer of 3'-thiacytidine, which is known to be a potent inhibitor of HBV replication. All of the patients but one were on cyclosporine-based immunosuppression. RESULTS: The mean serum creatinine was 134+/-44 micromol/L. The mean duration of HBV infection was 230+/-54 months (156 288). All of the patients but one had high serum alanine aminotransferase levels (122+/-52 IU/L; range, 45-243). Histological evaluation showed the presence of either chronic active hepatitis (n=4) or cirrhosis (n=2). All of the patients but one were hepatitis B e antigen negative/hepatitis B e antibody positive, but none were coinfected with either hepatitis C virus or hepatitis D virus. CONCLUSIONS: Lamivudine therapy was associated with (i) a normalization of alanine aminotransferase levels in four of five patients when these levels were increased at the beginning (n=5); (ii) a rapid disappearance of HBV DNA from the serum (detected by hybridization) in all of the patients; (iii) the negativity of HBV DNA by polymerase chain reaction in four patients; and (iv) no change in renal function and in proteinuria when present (one patient). Finally, no adverse effects were noted. When lamivudine therapy was stopped for four patients after 6 months, it was associated with a biochemical and virological relapse within the weeks that followed. Lamivudine therapy was therefore resumed for these patients. PMID- 9415573 TI - Limited T-cell repertoire in renal allograft and allogeneic melanoma transmitted by the graft. AB - In a patient with metastatic melanoma transmitted by the renal allograft, HLA serves as an alloantigen per se and is associated with tumor antigens at the same time. The influence of this antigeneic pattern on the Vbeta T-cell repertoire in an allogeneic melanoma, allograft, and peripheral blood mononuclear cells (PBMC) was assessed by polymerase chain reaction. Vbeta13.1 and 19 were found in both the melanoma and the graft. Vbeta14 was detected only in the melanoma and Vbeta6 was detected only in the kidney. PBMC revealed an unrestricted Vbeta pattern. Markers for cytotoxic activity of T cells--granzyme B and perforin--were not expressed during immunosuppressive therapy as clinically reflected in a nonrejecting allograft and in a progressing melanoma. In vitro PBMC proliferated to recombinant interleukin-2, whereas recombinant interferon-gamma did not augment this response. Initiation of immune therapy, in addition to discontinuation of immunosuppression, might support the rejection of the allogeneic tumor by dominant Vbeta T cells. PMID- 9415574 TI - Alopecia as a consequence of tacrolimus therapy in renal transplantation? PMID- 9415575 TI - Apoptosis is the main mechanism of cardiomyocyte death in the hyperacute rejection of heart xeno- and allografts. PMID- 9415576 TI - A review of probability of causation and its use in a compensation scheme for nuclear industry workers in the United Kingdom. AB - The assumption that any additional exposure to ionizing radiation leads to an increase in the risk of stochastic health effects implies that some cases of these effects will be caused by exposure incurred occupationally. The main health effect expected to arise in the exposed individuals is cancer. Such radiation induced cancers cannot be distinguished from the far larger number of background cancers, and, therefore, causation must be assessed statistically. The probability of causation methodology has been developed to ascertain the likelihood that a particular cancer may be attributed to a particular prior exposure to radiation. Given the pertinent details of an individual case, a probability of causation value is calculated from the appropriate relative risk obtained from radiation risk models derived from the epidemiological study of exposed populations, although there are many uncertainties inherent in a particular probability of causation calculation. In the United Kingdom, the Compensation Scheme for Radiation-linked Diseases has been created to determine whether those individuals occupationally exposed to radiation in the nuclear industry who have subsequently developed a malignant disease should be compensated. The Scheme is a voluntary arrangement based upon the probability of causation methodology, which incorporates various procedures agreed by employer and employee representatives and their advisers. In a pragmatic approach to compensation, the uncertainties of a specific probability of causation calculation are accommodated through generosity factors which favor the claimant and encourage the use of the Scheme. The Scheme, which was introduced in 1982 and modified in 1987 and 1991 in the light of operational experience and revised risk estimates, has provided a successful alternative to litigation from the point of view of both employer and employee. PMID- 9415577 TI - Modifiers of lung cancer risk in uranium miners from the Colorado Plateau. AB - Given the scientific consensus that exposure to radon decay products causes lung cancer, most recent studies have focused on the nature of the exposure-response relationship. Since residential radon exposure is now a primary public health issue, a better understanding of the effects of low levels of radon as well as factors modifying risk estimates has become very important. Several factors are shown to affect risk estimates in the latest update of the vital status follow-up (through 1990) and smoking history for the cohort of underground uranium miners in the Colorado Plateau. This analysis confirms earlier results indicating a strong dependence of relative risk estimates upon attained age. Quantitative estimates of relative risk as a function of cumulative exposure to radon decay products (WLM) are provided for three age strata. The non-linearity often reported in the Colorado Plateau data is shown to be at least partially due to an inverse exposure-rate effect, i.e., low exposure rates for long periods are more hazardous than equivalent cumulative exposure received at higher rates for shorter periods of time. However, this effect is shown to diminish at lower exposure rates and cumulative exposures. In addition, use of the new smoking data indicates that the radon/smoking interaction is submultiplicative and may depend upon attained age. PMID- 9415578 TI - Accounting for errors in dose estimates used in studies of workers exposed to external radiation. AB - This paper discusses approaches for accounting for errors in dose estimates used in dose-response analyses of data from epidemiologic studies of workers exposed to external radiation and illustrates these approaches with analyses of data on workers at the Hanford site. In these analyses, estimates of the excess relative risk are corrected for bias in recorded doses as estimates of organ dose, and confidence intervals reflect uncertainty in the correction factors. For the Hanford data, these procedures did not greatly modify results for all cancer excluding leukemia, but the upper confidence limit for leukemia was increased by about 40%, a difference that is of some importance in comparing worker-based estimates and confidence intervals with estimates that serve as the basis of radiation protection standards. It is argued that aside from taking account of uncertainty in correction factors, no additional corrections are needed to address random errors resulting from variation in exposure energies and geometries. In addition, it is shown that because the larger cumulative doses, which are most influential in dose-response analyses, are the sums of large numbers of independent dosimeter readings, random errors resulting from variation in laboratory measurements are unlikely to be important for epidemiologic purposes. It is hoped that the approaches illustrated in this paper will be applied to future analyses of data from worker studies, especially combined analyses of data from several countries. Taking account of uncertainty in factors that correct for systematic bias will be especially important as uncertainty resulting from sampling variation decreases. PMID- 9415579 TI - Monte Carlo simulation of in vivo measurements of 90Sr + 90Y bremsstrahlung. AB - The purpose of this study was to calculate the 90Sr + 90Y bremsstrahlung yield generated in bone quantitatively, and to determine the optimum conditions required for in vivo measurements of the yield, assuming that a given amount of contamination, due to the ordinary activities of 137Cs and 40K, is present within the human body. Several approximations in the distribution of activities in various body phantoms, simulating the human leg, waist and head, were made for simplicity. All the calculations were carried out using an in-house Monte Carlo code. The photon spectra incident on a detector window, of diameter 200 mm, positioned 1 mm from the surface of each phantom were obtained. A pseudo-peak of 90Sr + 90Y bremsstrahlung was observed at approximately 50 keV with a full width at half maximum of 60 keV (FWHM). The signal-to-background ratio, eta, where eta represents the ratio of the number of photons in the energy interval between 30 to 160 keV in the bremsstrahlung spectrum produced by 1 Bq kg(-1) of 90Sr and the background spectrum, which is assumed to comprise of 1 Bq kg(-1) of 137Cs and 60 Bq kg(-1) of 40K, was evaluated in order to determine the optimum measuring conditions. An optimum eta value of 0.0353 was obtained for the leg phantom, of bone diameter 30 mm, covered by a 1 mm layer of soft tissue. Combining the present calculations with the conventional measurements, a new and simpler method for evaluating the body burden activity of a beta emitter, such as 90Sr, is proposed. PMID- 9415580 TI - Neutron fluence and energy spectra around the Varian Clinac 2100C/2300C medical accelerator. AB - We have simulated the head geometry of a Varian Clinac 2100C/2300C medical accelerator in a Monte Carlo calculation to produce photoneutrons and transport them through the head shielding into a typical therapy room (modeled by a test cell at Varian Associates). The fast neutron leakage fluence and energy spectra have been calculated at 7 positions around the linac head for typical beam operation at 10, 15, 18 and 20 MV. The results of these calculations have been compared with limited measurements made using the same model accelerator operating in a Varian test cell. Calculations were also made for the fluence and energy spectra outside the head with no surrounding concrete walls, floor or ceiling to eliminate the effects of scattering from concrete. Comparisons were also made with calculations using a much simplified head geometry. The results indicate that the calculations using the complex head geometry compare, within the uncertainties, with the measurements. The simple head geometry leads to differences of a factor of 2 from the complex geometry. Results of these calculations can be used to calculate fast neutron transmission through various shielding configurations and through labyrinths. PMID- 9415581 TI - Combined diffusive and advective transport of radon in a homogeneous column of dry sand. AB - To validate a model for radon transport in soil, measurements of combined advective and diffusive radon transport under well-defined and controlled conditions have been made in a homogeneous column of dry sand with and without an air-filled volume on top. This volume simulates a crawl space. The measurements concern steady-state as well as time-dependent combined advective and diffusive transport and are performed in the framework of a systematic validation study which will also cover more complex situations (e.g., presence of pore-water). The experimental data were compared with results of a two-dimensional numerical model based on the finite-difference approach of the differential equation describing radon transport. Validations of the numerical model with analytical solutions were carried out for one-dimensional steady-state combined diffusive and advective transport, and for two-dimensional time-dependent diffusive transport. The results of the measurements with combined advective and diffusive transport are in good agreement with (numerical) model calculations with maximum deviations <10%. PMID- 9415582 TI - Analysis of quasicontinuous radon monitor response. AB - Continuous radon monitors respond quickly to changes in radon concentration but bear uncertainties due to the fact that the radon concentration in the detector changes within a counting interval. This disadvantage can be overcome by applying quasicontinuous sampling, where the detector chamber is sealed after the sample has been drawn, so that the radon concentration does not change while counting. The chamber is usually kept closed for 15 min or longer and subsequently flushed or evacuated before the new sample is taken. This paper presents a forward marching algorithm suitable for the analysis of the response of quasicontinuous radon monitors. The method can be used to obtain both 222Rn and 220Rn concentrations in the gas sample. PMID- 9415583 TI - A multi-user real time inventorying system for radioactive materials: a networking approach. AB - A computerized system for radioisotope management and real time inventory coordinated across a large organization is reported. It handles hundreds of individual users and their separate inventory records. Use of highly efficient computer network and database technologies makes it possible to accept, maintain, and furnish all records related to receipt, usage, and disposal of the radioactive materials for the users separately and collectively. The system's central processor is an HP-9000/800 G60 RISC server and users from across the organization use their personal computers to login to this server using the TCP/IP networking protocol, which makes distributed use of the system possible. Radioisotope decay is automatically calculated by the program, so that it can make the up-to-date radioisotope inventory data of an entire institution available immediately. The system is specifically designed to allow use by large numbers of users (about 300) and accommodates high volumes of data input and retrieval without compromising simplicity and accuracy. Overall, it is an example of a true multi-user, on-line, relational database information system that makes the functioning of a radiation safety department efficient. PMID- 9415584 TI - Modeling the migration of fallout radionuclides in soil using a transfer function model. AB - A stochastic model for transport of radionuclides in soil is presented. It is based on probability density functions of solute displacements, which are interpreted as impulse response functions of a linear dynamic system. Two transport models are discussed: (1) a convective-stochastic approach which takes into account spatial variability of flow and sorption and leads to a lognormal probability distribution of displacements, and (2) the conventional convective dispersive model with a constant retardation coefficient. To compare their applicability, both models were applied to depth distributions of 90Sr and 137Cs fallout measured in an Orthic Podsol. The convective-stochastic approach was found to provide a better representation of the observed depth distributions. Using this model, migration rates of the radionuclides were calculated. PMID- 9415585 TI - Uranium in urine--normalization to creatinine. AB - "Spot samples" of urine are routinely used to monitor occupational exposure to uranium and other toxic heavy metals, such as mercury, lead, and cadmium. In the present work, it was shown that diurnal variations in the uranium concentration in different urine samples from the same individual could be quite large. However, these variations were in correlation to the creatinine level of the same samples, with values of R = 0.72-0.99, for the five subjects studied here. Thus, it is proposed here that uranium concentrations in "spot" urine samples be expressed in terms of ng uranium g(-1) creatinine rather than ng uranium L(-1). Once the 24-h creatinine level is estimated for the individual based on weight, height and age, the adjusted values can be used for determination of the internal dose of uranium. PMID- 9415586 TI - Measurement of 222Rn concentration in Kenyan groundwater. AB - Groundwater samples from different parts of Kenya have been analyzed for their 222Rn concentrations. Samples were drawn from municipal supplies, springs, wells and bore holes in different geological terrains. In the analysis, conventional liquid scintillation counting, using the Packard protocol for measuring radon in water, was employed. This is the first time these water sources are being tested for radioactivity. The preliminary results are presented, and they showed that 222Rn is the main contributor to the radioactivity in most portable waters in Kenya. 222Rn activity concentrations range from 0.8 +/- 0.5 to 371.7 +/- 33.5 Bq L(-1). PMID- 9415587 TI - Evidence for photoneutron production in the lead shielding of a dedicated intra operative electron only facility. AB - A dedicated electron-only intra-operative suite has just been completed at the Massachusetts General Hospital. This suite is located on the 3rd floor with the shielding consisting entirely of lead and borated polyethylene, except for concrete in the floor and ceiling to support the lead and on the finished floor. The radiation protection barriers for this facility were calculated on the basis of a maximum permissible dose of 10 microSv wk(-1) for photons and 10 microSv wk( 1) for neutrons, based on a quality factor of 10 (this factor is specified in the regulations of the Commonwealth of Massachusetts). This even split was predicated on the basis that, except for the primary beam, the neutron leakage from the machine was generally about the same as for x rays. The initial survey showed that the neutron dose equivalent outside all barriers, except below the floor, was within the calculated values, < 10 microSv wk(-1). However, the neutron dose equivalent measured directly below the floor was found to be 35 microSv wk(-1) for the highest electron energy, a difference of > 25 microSv wk(-1). The neutron dose equivalent, for neutrons produced by bremsstrahlung photon interactions in the lead and based on a 3% photon background in the electron beam, was calculated to be 25 microSv wk(-1), in good agreement with this difference. PMID- 9415588 TI - Radioactive contamination incidents involving protective clothing. AB - The study focuses on incidents at Department of Energy facilities involving the migration of radioactive contaminants through protective clothing. The authors analyzed 68 occurrence reports for the following factors: (1) type of work; (2) working conditions; (3) type of anti-contamination material; (4) area of body or clothing contaminated; and (5) nature of spread of contamination. A majority of reports identified strenuous work activities such as maintenance, construction, or decontamination and decommissioning projects. The reports also indicated adverse working conditions that included hot and humid or cramped work environments. The type of anti-contamination clothing most often identified was cotton or water-resistant disposable clothing. Most of the reports also indicated contaminants migrating through perspiration-soaked areas, typically in the knees and forearms. On the basis of their survey, the authors recommend the use of improved engineering controls and resilient, breathable, waterproof protective clothing for work in hot, humid, or damp areas where the possibility of prolonged contact with contamination cannot be easily avoided or controlled. PMID- 9415590 TI - Specific activity. PMID- 9415589 TI - An international intercomparison of soil gas radon and radon exhalation measurements. AB - The Environmental Measurements Laboratory hosted the Sixth International Radon Metrology Programme Intercomparison Test and Workshop (IRMP6) from 12-15 June 1995. Thirty participants representing 24 different institutions from 11 countries attended. Laboratory exercises consisted of 220Rn and 222Rn concentration measurements from a source container, and exhalation measurements from a 226Ra-spiked concrete slab and a "normal" concrete slab. Field exercises included soil gas radon measurements and radon exhalation measurements. In this report, we pooled the participants' data and used the ratio of the standard deviation (SD) to the arithmetic mean, expressed as a percentage, to assess participant agreement for each exercise. For the exhalation measurements from the 226Ra-spiked slab, this value is 37%; for soil gas 222Rn, this value is 120%, 36% and 27% for each depth range, 0.4-0.5, 0.6-0.75 and 0.9-1.0 m, respectively; for the surface exhalation measurements, this value is 34%. For the drum 222Rn measurements, the percent SD after removing a linear trend was 13%. These results indicate that sampling errors are greater than instrument errors. PMID- 9415591 TI - Response to Wolbarst and Mauro. PMID- 9415592 TI - Oceanic disposal of radioactive waste: science vs. politics. PMID- 9415593 TI - Endoscopic laryngotracheoplasty and graft soldering with the carbon dioxide laser. An animal study. AB - Laryngotracheoplasty (LTP) has had considerable success for treatment of selected cases of subglottic stenosis in children. However, certain complications have been associated with an open approach. Our study explores the possibility of decreasing these complications by means of a minimally traumatic endoscopic laser cricoid split and graft transplantation technique. In an in vivo study of six dogs, LTP was performed with a carbon dioxide laser. The gap between the divided cricoid cartilage was either left with no coverage or filled by autogenous mucosa or cartilage grafts through a laser-assisted soldering technique. Compared to LTP without coverage, autogenous grafts, with either mucosa or cartilage, resulted in improved healing. The cartilage graft seems to offer the best result for expansion of the subglottic space. The results show that the new intraluminal approach, a combination of an endoscopic cartilage split and graft transplantation, may be an alternative to standard LTP. The advantage of this technique is that it is less traumatic than the open procedure, with the potential for less morbidity. PMID- 9415594 TI - Telescopic laryngeal and pharyngeal surgery. AB - Surgery of the hypopharynx and larynx has traditionally been performed with either direct, unaided vision or the operating microscope. We proposed to extend the surgical capability provided by angled Hopkins telescopes to the larynx and hypopharynx. Forty-eight cases in which rigid telescopes were employed primarily for intervention were reviewed. We found significant advantages of the telescopic system when performing procedures on surfaces that were not 90 degrees from the observer, such as the walls of the hypopharynx, the petiole of the epiglottis, the ventricle, and the posterior commissure. Both 30 degree and 70 degree telescopes were found useful, but required complementary instruments. The potassium titanyl phosphate laser's flexible fiber provided a distinct advantage in resecting lesions that presented on vertical surfaces. Telescopes also permitted the use of large instruments designed for intraperitoneal and intrathoracic surgery that blocked the view of the operating microscope. Telescopic pharyngeal and laryngeal visualization allowed surgical procedures complementary to more traditional forms of endoscopic surgery. PMID- 9415595 TI - Laryngotracheoesophageal clefts. AB - This article reviews laryngeal cleft anomalies from the Laryngeal Development Laboratory at Children's Memorial Hospital in Chicago and includes a discussion of the classification of laryngotracheoesophageal clefts based on previous work and the information presented herein. Of the 115 laryngeal specimens obtained between 1975 and 1995, 11 have laryngeal cleft anomalies. Eight have a submucous laryngeal cleft. There is 1 laryngotracheoesophageal cleft, type II (partial cricoid cleft); and there are 2 laryngotracheoesophageal clefts, type III (complete cricoid cleft). The histopathologic findings are presented in detail and the literature is reviewed. Photomicrographs and drawings illustrate the pathology and classification. Clinical presentation, diagnosis, evaluation, and management are discussed, as is the embryology. PMID- 9415596 TI - Efficacy of repeated botulinum toxin injections as a function of timing. AB - This pilot study was designed to determine if the interval between repeated botulinum toxin injections influenced physiologic and histologic effects on laryngeal muscles in a rat model. The physiologic measurements included digitized videomicroscopic recording of vocal fold movement and electromyography. The histologic measurements included muscle fiber size and digitized optical density of laryngeal muscles after glycogen depletion by electrical stimulation. The results demonstrated that the effect of timing of the second injection was strongly correlated to laryngeal changes. Most notable were results in the subjects that underwent injections 6 weeks apart. We hypothesize that these findings might be related to terminal axonal sprouting with reinnervation. The results from this study help confirm and expand the validity of using the rat laryngeal model to understand the effect of botulinum toxin. Moreover, we believe that the data might be extrapolated to prove useful in predicting human responses to botulinum toxin treatment for functional dystonias such as spasmodic dysphonia. PMID- 9415597 TI - Utility of helical computed tomography in the study of arytenoid dislocation and arytenoid subluxation. AB - Conventional computed tomography (CT) has been considered a mainstay in the evaluation of the larynx. A major difficulty with utilizing this modality, especially in the study of the arytenoid, is the time necessary to perform a thin slice examination through a structure that has a propensity to move with respiration and swallowing. Helical CT not only significantly reduces the time necessary to study the larynx, but enables one to perform multiple high resolution multiplanar reconstructions. Eleven patients with arytenoid abnormalities documented by strobovideolaryngoscopy or direct laryngoscopy were imaged with helical CT. A comprehensive radiographic examination illustrating the cricoarytenoid relationship in all of the subjects was completed in less than 20 seconds by using axial reconstructions in 2-mm-thick slices at 1-mm intervals, with subsequently derived sagittal and coronal reconstructions. Helical CT may be a useful adjunct in the diagnosis of arytenoid subluxation or dislocation. PMID- 9415598 TI - Basaloid squamous cell carcinoma of the larynx and hypopharynx. AB - Basaloid squamous cell carcinoma (BSCC) is a recently described bimorphic variant of squamous cell carcinoma with distinct morphological and biological features. We describe the clinicopathological findings, along with immunohistochemical and ultrastructural investigations, in 15 new cases of BSCC of the larynx or hypopharynx observed and treated at the otolaryngology department of the University of Padua between 1989 and 1995. The world literature is also reviewed in order to develop a more accurate clinicopathological profile of the tumor. Patient records and histologic slides were reviewed in all of our 15 cases. The patient group consisted of 13 men (86.67%) and 2 women with a mean age of 63.33 years (median 69 years; range 44 to 84 years). Nine patients presented with cervical lymph node metastases. Surgical treatment was the therapy of choice; radiotherapy and chemotherapy have been applied in different combinations. Follow up was available on all 15 cases. Local recurrence was described in 3 cases. Five of the 9 patients with cervical lymph node metastases developed distant metastases. Distant spread of the tumor without lymph node involvement was observed during follow-up in 4 cases. Nine patients died of disease, 2 are alive with widespread metastases, 2 are alive with no evidence of disease, and 2 have died of other causes. The determined 5-year survival was estimated to be 17.5% by the Kaplan-Meier method. In conclusion, BSCC is a distinctive carcinoma that is important to recognize, because it has a more aggressive biological behavior than conventional squamous cell carcinoma. PMID- 9415599 TI - Comparison of carbon dioxide and neodymium: yttrium-aluminum-garnet lasers in surgery of the inferior turbinate. AB - At the Department of Otorhinolaryngology-Head and Neck Surgery of the University of Kiel, 533 patients with hyperplastic inferior turbinates were treated between 1987 and 1994 with various carbon dioxide (CO2) and neodymium:yttrium-aluminum garnet (Nd:YAG) laser techniques. We report on the therapeutic results of both types of laser turbinectomy and compare their long-term results with those of submucosal diathermy. Among the different techniques, we preferred the following approaches. The CO2 laser technique involved the application of a few laser spots (laser energy density 6,100 J/cm2 per lesion) to the head of the turbinate under the operating microscope. In the Nd:YAG laser procedure, diffuse, low-power irradiation (laser energy density < 53 kJ/cm2) of the entire concha was performed under endoscopic control. The CO2 laser procedure involved little bleeding and hardly any pain. It produced a positive effect after only a few days and required no follow-up treatment. The success of Nd:YAG laser treatment, by contrast, only became evident after weeks or months, due to the slow scarring process. Compared to submucosal diathermy, both laser methods produced better long-term results. Two years postoperatively, the overall success rate, as defined by patient satisfaction, was 79.6% for the CO2 laser, 68.3% for the Nd:YAG laser, and 36% for submucosal diathermy. PMID- 9415600 TI - Algorithm analysis of lectin glycohistochemistry and Feulgen cytometry for a new classification of nasal polyposis. AB - The aim of this study is to present a new classification of nasal polyps. This classification is based both on morphologic criteria relating to morphonuclear features from isolated Feulgen-stained nuclei and on glycohistochemical characteristics from histologic slides submitted to three lectins (peanut, wheat germ, and gorse seed agglutinins) and one neoglycoconjugate glycohistochemical stain. While the morphonuclear features (including 30 variables) relate essentially to chromatin pattern, the glycohistochemical stains (including 16 variables) are linked to the presence of specific carbohydrate moieties in cell membranes and cytoplasm. Forty-nine nasal polyps, including single polyps, diffuse polyposis, cystic fibrosis-related polyposis, and aspirin idiosyncracy related polyposis associated with asthma, were thus characterized. All the variables were obtained quantitatively by means of computer-assisted microscopy. Two complementary methods of data classification were used to determine the actual diagnostic value contributed by each quantitative variable, namely, discriminant analysis, which forms part of multifactorial statistical analysis, and the decision tree technique, which is an artificial intelligence-related algorithm. The data so obtained show that our morphologic classification of nasal polyps fits in with the classification of nasal polyps defined on the basis of clinical criteria. PMID- 9415601 TI - Statistical evaluation of hearing screening by distortion product otoacoustic emissions. AB - The aim of this study was to provide a statistical evaluation of the screening properties of distortion product otoacoustic emissions (DPOEs) in individuals with clinically normal hearing and in patients with pure sensorineural deafness of various degrees. The main informational parameters used were sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and the analysis of receiver operating characteristic (ROC) curves. For each frequency tested, ears were classified as a function of their audiometric threshold. Two groups were defined relative to an arbitrary reference, the "audiometric criterion." The PPV decreased and NPV increased with increases in the audiometric criterion. Each point of the ROC curve represents the relationship between the false alarm rate and the hit rate for each audiometric criterion ranging between 10 and 75 dB hearing level: the lower the audiometric criterion, the lower the hit rate value, and the lower the false alarm value. The audiometric criterion giving the highest hit rate and the lowest false alarm rate was 55 to 60 dB hearing level for primaries at 60 and 70 dB sound pressure level, or 25 to 30 dB hearing level for primaries at 30, 40, and 50 dB sound pressure level. These two different behaviors of ROC curves are consistent with the hypothesis that DPOEs do not represent activity at a single location along the basilar membrane. PMID- 9415602 TI - Three familial cases of hearing loss associated with enlargement of the vestibular aqueduct. AB - The present report describes three familial cases of recessive hearing loss associated with enlargement of the vestibular aqueduct (EVA). Six siblings from three families showed EVA. The common characteristic of these patients was the presence of congenital, high-frequency, fluctuating sensorineural hearing loss. These cases suggest that EVA may be a useful discriminator between different types of recessive hearing loss. PMID- 9415603 TI - Reduction of inner ear inflammation by treatment with anti-ICAM-1 antibody. AB - The in vivo effect of systemic administration of monoclonal antibody (mAb) against ICAM-1 (alphaICAM-1) in experimental immune-mediated labyrinthitis was evaluated. The alphaICAM-1-treated rats showed reduced inflammatory cell infiltration in the scala tympani and the perisaccular tissue of the endolymphatic sac. However, with this experimental paradigm, labyrinthitis could not be completely abolished. These findings suggest that ICAM-1-dependent pathways play an important role in the series of immunologic events occurring in the inner ear, and that the use of ICAM-1 antagonist may be a possible therapeutic approach to labyrinthitis. PMID- 9415604 TI - Changes in myosin expression in denervated laryngeal muscle. AB - The effects of chronic denervation on the myosin heavy chain (MyHC) content and muscle fiber type composition of rat laryngeal muscles are described. The posterior cricoarytenoid (PCA) and thyroarytenoid (TA) muscles were removed 3 weeks, 3 months, and 6 months after recurrent laryngeal nerve sectioning. Myofibrillar adenosine triphosphatase staining of cryostat sections was performed, and fiber type percentages were determined. Sodium dodecyl sulfate polyacrylamide gel electrophoresis was used to separate MyHC isoforms, and densitometry was subsequently used for quantitative analysis. Unoperated animals served as controls. In the PCA muscle, denervation resulted in a progressive reduction in type I MyHC (the slow-contracting isoform) to an almost complete loss at 6 months, with a concomitant increase in type II MyHCs (fast-contracting isoforms, excluding type IIL). Type IIL MyHC (laryngeal-specific isoform) remained relatively constant up to 6 months after denervation. The myosin expression in the TA muscle, which contained only type II MyHCs, remained relatively constant with denervation. Changes in fiber type composition of the muscles described from tissue staining correlated with MyHC content. These findings in laryngeal muscle confirm the dependence of type I MyHC expression upon neural input, as has been found previously in limb skeletal muscles. Since the expression of all MyHCs except the IIL was modified after denervation in the PCA muscle, it is possible that the IIL isoform is maintained by factors that differ from those in the other skeletal myosins. PMID- 9415605 TI - Transection and occlusion of lateral semicircular canal in guinea pigs. AB - Audiologic and histopathologic examinations were made after occlusion of the lateral semicircular canal in eight guinea pigs. The lateral semicircular canal was drilled out, and then the canal lumens were plugged with muscle pieces. After a serial recording of auditory brain stem responses for 2 months, histologic specimens of the temporal bones were prepared in the lateral semicircular canal plane. One animal developed profound hearing loss due to suppurative labyrinthitis. The other seven animals showed no significant threshold elevation during this period. Histopathologic examination revealed that the bone defect on the lateral canal was replaced with newly formed bone; the perilymphatic and endolymphatic spaces maintained their compartmentalization; and the membranous endolymphatic canal healed to form complete blind ducts. These findings suggest that proper management of the injured semicircular canal is important for maintenance of postoperative hearing. PMID- 9415606 TI - Auditory evoked responses under total spinal anesthesia in rats. AB - In order to investigate the function of the auditory pathway from the cochlea to the brain stem under total spinal anesthesia, the auditory brain stem response (ABR), compound action potential of the cochlear nerve (CAP), and cochlear microphonics (CM) were simultaneously recorded in rats. Total spinal anesthesia was induced by infusion of 2% lidocaine hydrochloride at a constant rate of 0.10 mL/min into the cerebrospinal fluid through the rats' skulls. The ABR completely disappeared within 1.5 to 4 minutes. After cessation of the injection, the ABR reappeared, starting from wave I and progressing through waves II and III to wave IV. The latency change of the CAP throughout the recording period was quite similar to that of wave I of the ABR. A reduction in amplitude of the CM was observed, but the CM did not disappear during the recording period. Disappearance of the ABR was due, not to loss of cochlear function, but to anesthetic effects on the acoustic nerve and the brain stem. Monitoring of the ABR provided information on the level of neural activity in the brain stem under total spinal anesthesia. PMID- 9415607 TI - Iatrogenic pseudoaneurysm of the internal carotid artery. PMID- 9415608 TI - Imaging diagnosis of buckled innominate artery. PMID- 9415609 TI - Ethmoid and sphenoid mucoceles viewed by helical scanning computed tomography with sagittal reconstruction. PMID- 9415610 TI - Evaluation of classic architectural criteria in non-mycosis fungoides cutaneous lymphomas. AB - Ninety-seven cases of non-mycosis fungoides (non-MF) cutaneous lymphoma were evaluated employing published criteria for the categorization of B- and T-cell cutaneous malignancies. Included in the study were 77 primary and secondary cutaneous B-cell lymphomas, in which the diagnosis was supported by immunohistochemical studies identifying lineage. These cases were randomized with 20 cases of non-MF and T-cell lymphoma. Hematoxylin and eosin (H & E)-stained slides from each case were reviewed by at least two dermatopathologists, who were unaware of the previous diagnoses, and a judgment regarding histologic pattern was rendered. The histologic criteria employed emphasized architectural features. For B-cell patterns, these included the presence of dense perivascular, periappendageal and/or nodular collections of lymphocytes, centering in the deep dermis, and separation from the epidermis by a grenz zone. Employed criteria for cutaneous T-cell pattern included location restricted primarily to the upper dermis, interstitial pattern, the presence of epidermotropism, and the lack of a grenz zone. Three B-cell lymphomas were judged to have indeterminate patterns. Four of 74 (5.4%) of the remaining B-cell lymphomas were incorrectly categorized as T-cell lymphomas by architectural criteria. The most striking findings included epidermotropism in rare B-cell lymphomas. Three of the four miscategorized cases were large-cell lymphomas. A preference for B-cell pattern was also confirmed in non-MF T-cell lymphomas. We conclude that most B-cell lymphomas in the skin demonstrate a recognizable B-cell pattern, but rarely a pattern more reminiscent of T-cell lymphoma may be seen. This may occur more often with B-large-cell lymphomas. In addition, this study supports previous work indicating that many non-MF T-cell lymphomas show prominent architectural features typically ascribed to B-cell lymphomas. In summary, our findings support the impression that the vast majority of non-MF lymphomas show a B-cell pattern regardless of their lineage. As such, caution is indicated with regard to pattern interpretation. PMID- 9415611 TI - Dermatofibrosarcoma protuberans with fibrosarcomatous areas: a clinico-pathologic and immunohistochemic study in four cases. AB - Dermatofibrosarcoma protuberans (DFSP) with fibrosarcoma (FS)-like areas (DFSP FS) is a peculiar neoplasm that combines microscopic findings of DFSP and FS. Because of the scarce number of cases published, tumor prognosis remains controversial. The clinical histories and the histologic material of 27 cases of DFSP were reviewed. Four of them showed fibrosarcomatous areas. Follow-up data, ranging from 12 to 125 months, were obtained in all four cases. Two patients had repeated local recurrences. One patient developed pulmonary metastases and died of disease 49 months after diagnosis. In the other two patients, no recurrences or metastases were detected at 12 and 70 months after local excision, respectively. Progressive increase of FS areas, cellular density, cellular atypia, and mitotic activity were observed during the recurrences. All cases showed diffuse positive immunostaining for CD34 in DFSP areas. Three cases were also CD34-positive in FS areas. Based on a careful review of the literature and our personal experience, we conclude that DFSP-FS is a rare variant of DFSP with a higher rate of local recurrences and more distant metastases than typical DFSP. PMID- 9415612 TI - Basal cell carcinoma with thickened basement membrane: a variant that resembles some benign adnexal neoplasms. AB - Because cutaneous basal cell carcinoma (BCC) is such a common malignancy, its unusual histologic manifestations are important. We identified a variant of BCC in which thickened basement membranes surround aggregations of neoplastic epithelial cells. Thickened basement membranes of similar appearance have previously been observed in benign cutaneous adnexal neoplasms, in basaloid monomorphic adenomas of the salivary gland and in other benign conditions, such as folliculocentric basaloid proliferation. We identified nine BCCs that otherwise met standard criteria, but which also had thick basement membranes surrounding some of the aggregations, and examined them by routine and histochemical staining. The cases included BCC with nodular, micronodular, and infiltrating patterns. Two neoplasms were composed largely of clear cells, suggesting, together with the thickened membranes, outer root sheath differentiation. CD34, which labels keratinocytes of the outer root sheath, marked only the epithelial cells of one of these cases. The thickened membranes were stained by periodic-acid Schiff with and without diastase (PAS-D) and by antibodies to type IV collagen and laminin, with slightly different staining patterns. Intraepithelial droplets within aggregations stained with PAS-D and type IV collagen antibodies. Thickened basement membranes therefore can occur in most of the common growth patterns of BCC. The absence of CD34 staining of epithelial cells in most cases makes it problematic at this time to prove that the thickened membranes indicate trichilemmal differentiation. BCC with thick basement membranes can closely mimic benign neoplasms, such as cylindroma and trichilemmoma, from which they can be distinguished in routinely stained sections. The presence of a continuous thick basement membrane around aggregates of epithelial cells does not in and of itself distinguish between benign and malignant cutaneous epithelial neoplasms. PMID- 9415613 TI - Basal cell carcinomas arising over arteriovenous malformations: some speculations on the theme. AB - The modulating effects of mesoderm on ectoderm during the embryonic period have been well documented. Some cutaneous conditions such as dermatofibroma with basaloid proliferation are examples of such a relationship. Seven cases of basal cell carcinoma (BCC) developing in port-wine stains and one case developing over an arteriovenous (AV) malformation have been reported. We report 3 elderly patients with BCC developing over AV malformations of the lower extremities. The association between BCC and AV malformation range from a chance association to a complex interaction between mesenchyme and epithelium, in which mesenchyme induces and modulates epithelial growth through the elaboration of specific growth factors (GF). PMID- 9415614 TI - HMB45 negative spindle cell malignant melanoma. AB - Desmoplastic malignant melanoma (DMM) and spindle cell malignant melanoma (SCMM) form a continuum without a discrete separation. One feature characteristic of DMM is a negative reaction for HMB45, a marker for premelanosomes. Fifty-six cases of SCMM were stained with HMB45. The clinical features, histologic features, and survival data for HMB45(+) and HMB45(-) SCMM were compared. Thirty cases were HMB45(-), and 26 were HMB45(+). In the HMB45(-) cases, there was a 1.4:1 ratio of males to females, and in the HMB45(+) cases the ratio was 1:1.5. Only 12.9% of the HMB45(-) cases occurred on the trunk compared with 40% of the HMB45(+) cases. The average ages for the HMB45(-) and the HMB45(+) cases were 65.6 and 61 years, respectively. Follow-up was obtained on 22 cases: 11 HMB45(-) and 11 HMB45(+). Of the 11 HMB45(-) cases, four had a 5-year disease-free survival. Of the 11 HMB45(+) cases, only one had a 5-year disease free survival. HMB45(-) SCMM appear to have a less aggressive biologic potential than HMB45(+) SCMM. PMID- 9415615 TI - In situ characterization of the human host response to Leishmania panamensis. AB - Both host and parasite determinants influence the outcome of Leishmania infections. Human host responses in cutaneous leishmaniasis of limited duration caused by a single species of the Viannia (V) subgenus were studied in skin biopsies obtained from lesions caused by Leishmania (V) panamensis in 31 male patients from the Colombian Pacific Coast. Dermal infiltrates and histopathologic changes were characterized using monoclonal antibodies and an indirect immunoperoxidase method. Dermal distribution of T-cell subpopulations and B lymphocytes was nonrandom: CD4+ and CD8+ T cells were most frequent in the upper dermis, and B cells were most abundant in the lower dermis. Parasites, macrophages, and neutrophils were localized predominantly in the middermis. Multiple regression analyses to establish associations between lesion type (ulcer, nodule, or papule), immune response data (Montenegro skin test, indirect fluorescence antibody test titers, lymphocyte blastogenesis), and particular cell populations demonstrated statistically significant correlations between CD4+ lymphocytes and macrophages (p < 0.05). CD8+ lymphocytes were associated with plasma cells (p < 0.001), as was the presence of amastigotes (p < 0.05). These associations and the in situ divergence of CD4 and CD8 ratios suggest that prognostic indicators for disease evolution could be identified by prospective analysis of cellular relationships and response to therapy. PMID- 9415616 TI - Adult T-cell leukemia/lymphoma associated with noninfectious epithelioid granuloma in the skin: a clinicopathologic study. AB - Hodgkin's disease (HD) and non-Hodgkin's lymphoma (NHL) are infrequently associated with noninfectious granulomas in involved or noninvolved organs. Adult T-cell leukemia/lymphoma (ATLL) is an aggressive lymphoproliferative neoplasm associated with T-lymphotropic virus type 1 (HTLV-1). We describe a case of cutaneous type ATLL, affecting mainly the skin as a maculopapular eruption, in which some skin biopsies contained epithelioid cell granulomas in the lymphoma cutis (ATLL) lesion. These Lennert's-like epithelioid clusters were also present in lymph nodes, which showed some degree of invasion by the ATLL lymphocytes. Although prognosis of ATLL is generally poor, our patient has had a less aggressive course, with a survival time to date of 13 years. Our findings suggest that the presence of epithelioid granulomata in an ATLL patient may be a manifestation of a host response which confers some protection against the disease progression. To our knowledge, this is the first report of a case of ATLL with a noninfectious granuloma similar to a Lennert's lesion. PMID- 9415617 TI - Primary papular xanthoma of children: a clinicopathologic, immunohistopathologic and ultrastructural study. AB - Papular xanthoma (PX) is a very rare skin disorder. We describe a typical case of PX in a 13-month-old Chinese boy who presented with numerous yellow-red papulonodules, 2-8 mm in diameter, mainly on the face, both upper extremities, and abdomen of 10 months duration. Histologic studies showed a diffuse monomorphous infiltrate of foamy cells in the upper dermis. The foamy cells stained positively with oil red O and CD68. The periodic acid Schiff (PAS) stain, S-100 protein, CD1a, CD56, lysozyme, alpha1-antitrypsin, and factor XIIIa were all negative in the foamy cells. The electron microscopic (EM) studies revealed the morphologic features of macrophages with electron-dense, membrane-limited lipid vacuoles in the cytoplasm. After 14 months, neither spontaneous regression nor anetoderma-like scars were noted. Our immunohistochemical and ultrastructural studies support the notion that the origin of the foamy cells is the macrophage rather than the factor XIIIa (+) dermal dendrocyte. There was no associated or underlying disease in this case. We suggest the term primary PX for cases such as this one. PMID- 9415618 TI - Fatal mural endocarditis and cutaneous botryomycosis after heart transplantation. AB - Fatal mural endocarditis and botryomycosis occurred concurrently in a 62-year-old women 4 months after orthotopic heart transplantation. Subsequent to mild mitral regurgitation, infection developed on a left atrial thrombus and was complicated by cerebral embolization. Simultaneously, skin nodules manifested on both forearms. Histologic examination revealed typical aspects of early evolving botryomycosis with massive infiltration of the dermis and hypodermis by necrotic granulomas framed by grains of Gram-positive coccoid forms. Bacteria were decorated by a nonspecific polyclonal antibody to Mycobacterium bovis. PMID- 9415619 TI - Angiomatoid melanoma: a case of metastatic melanoma mimicking a vascular malignancy. AB - We present a case of melanoma metastatic to forehead skin and sacral vertebra, without known primary, which, in the cutaneous metastasis, displayed striking histologic features suggesting vascular differentiation. Histopathologic examination of the lesion revealed a large, deeply extending spindle cell malignancy with numerous, cavernous, erythrocyte-filled spaces throughout and only scant melanin pigmentation, making initial diagnosis challenging. The neoplastic cells demonstrated positive staining with antibodies to S-100 protein, HMB 45, and vimentin. We suggest the term angiomatoid melanoma for this histopathologic variant. PMID- 9415620 TI - Primary cutaneous neuroendocrine (Merkel cell) carcinoma in association with squamous- and basal-cell carcinoma. AB - The co-existence of squamous-cell carcinoma (SCC) and neuroendocrine (Merkel cell) carcinoma (NEC) of the skin has been well documented in several patients. The concurrent development of NEC and basal-cell carcinoma (BCC) in the same cutaneous lesion, on the other hand, has not been reported. We describe a 68-year old man presenting with NEC, SCC, and BCC arising at the same site in retroauricular skin. Cells of NEC and SCC showed an intermingled pattern of growth, whereas those of BCC were "embedded" among those of the other two tumors. Immunohistochemical stains were consistent with the histopathologic diagnosis. We believe that this is the first report of the association of NEC, SCC, and BCC in the same skin lesion. PMID- 9415621 TI - Merkel cell carcinoma with partial spontaneous regression: an immunohistochemical, ultrastructural, and TUNEL labeling study. AB - We report a case of Merkel cell carcinoma that partially regressed after biopsy. A 76-year-old woman presented with an 1 month history of a rapidly enlarging nodule on her left cheek. After biopsy, the nodule reduced to almost half the size and was excised 1 month later. The excised specimen showed a dense cluster of lymphocytes and fibrosis around the tumor nests. In addition, lymphocytes showed apposition with tumor cells. An immunohistologically dense, even infiltration of CD4+ and CD8+ cells was found around the tumor nests, and more CD8+ cells than CD4+ cells were seen in the tumor nests. By electron microscopy (EM), apoptosis of tumor cells and lymphocytes was observed. Many apoptotic cells were also detected by in situ nick end-labeling (TUNEL) of DNA-breaks, especially in the marginal area of tumor nests surrounded by dense lymphocytic infiltrates. It seems likely that T-cell immunity, which induces apoptosis of tumor cells, may have been involved in tumor regression. PMID- 9415622 TI - Pigmented trichoblastoma. PMID- 9415624 TI - The hybrid epidermoid and apocrine cyst. PMID- 9415625 TI - p53 protein expression in Kaposi's sarcoma. PMID- 9415626 TI - Lupus and interleukin 10. PMID- 9415627 TI - Alternative therapies--medicine, magic, or quackery. Who is winning the battle? PMID- 9415628 TI - Alternative medicine--the challenge of nicht-Schulmedizin (or what is not taught in medical schools) PMID- 9415629 TI - Differences in the use of second-line agents and prednisone for treatment of rheumatoid arthritis by rheumatologists and non-rheumatologists. AB - OBJECTIVE: To compare the use of methotrexate (MTX), intramuscular (i.m.) gold, hydroxychloroquine, and prednisone for rheumatoid arthritis (RA) treatment among patients managed by rheumatologists and nonrheumatologists. METHODS: Multiple regression analysis to estimate the likelihood of starting treatment and response to treatment for patients managed by rheumatologists and nonrheumatologists. All regression analyses were adjusted for patient demographic and clinical characteristics. RESULTS: Therapy with all agents studied was initiated more frequently for patients with RA with at least some contact with rheumatologists during the year than for those managed strictly by nonrheumatologists. The adjusted odds ratios for starts on these medications ranged from 1.14 for im gold to 15.11 for MTX for patients managed by rheumatologists compared to those managed by nonrheumatologists. However, due to the low frequency of initiation of treatment with most of these drugs for patients managed strictly by nonrheumatologists, only the odds ratio for prednisone reached statistical significance (OR = 2.94, p = 0.0082). In the year after initiation of therapy with these agents, patients managed by rheumatologists experienced better response to treatment than those managed by nonrheumatologists. These differences were statistically significant for MTX (p = 0.0447) and nearly significant for im gold (p = 0.0597). CONCLUSION: These results provide evidence of systematic differences in the propensity of rheumatologists and nonrheumatologists to initiate therapy with these antirheumatic drugs. If the observed differences in initial response to treatment translate into substantial differences in longterm outcomes, then these results suggest that the welfare of patients with RA may be jeopardized by the current trend toward primary care and restricted access to rheumatologists. PMID- 9415630 TI - Variability in cytokine and cell adhesion molecule staining in arthroscopic synovial biopsies: quantification using color video image analysis. AB - OBJECTIVE: To investigate the variability in immunostaining for cytokines and cell adhesion molecules using multiple arthroscopically directed synovial biopsies from within a rheumatoid knee joint, quantitated by color video image analysis. METHODS: Needle arthroscopic biopsies were taken from multiple sites (4 7 sites) around a knee joint in 8 patients with rheumatoid arthritis (RA). In 5 patients, immunoperoxidase staining for the cytokines tumor necrosis factor alpha (TNF-alpha), interleukin 8 (IL-8), and IL-1beta as well as the IL-1 receptor antagonist protein (IL-1ra) was performed. In 3 patients, immunoperoxidase staining for the cell adhesion molecules E-selectin (CD62E), P-selectin (CD62P), intercellular adhesion molecule 1 (ICAM-1, CD54), and platelet endothelial cell adhesion molecule (PECAM, CD31) was performed. Immunostaining was quantified using color video image analysis. RESULTS: The overall probability of paired biopsies from the same RA knee joint being significantly different from each other due to sampling variation was at most 22% for cytokine staining (usually less than 10%). There were no significant differences between intrabiopsy and interbiopsy variability for cell adhesion molecule staining of the sublining and vessels. CONCLUSION: The variability in cytokine and cell adhesion molecule staining within any single biopsy usually reflects the variability between biopsies taken from different sites in the same rheumatoid joint when the immunostaining is quantified using color video image analysis. Therefore, only a small number of synovial biopsies are required to accurately determine the cytokine and cell adhesion molecule expression in a single joint. PMID- 9415631 TI - Clinical characteristics of patients with rheumatoid arthritis and methotrexate induced pneumonitis. AB - OBJECTIVE: To determine the clinical features of methotrexate (MTX) pneumonitis in patients treated for rheumatoid arthritis (RA). METHODS: The medical records of 284 patients with RA who had been treated with oral MTX (mean followup 33.2 mo) were reviewed retrospectively. RESULTS: MTX induced interstitial pneumonitis developed in 6 patients (2.1%). The affected patients were significantly older than those without MTX pneumonitis (67.3 +/- 9.8 vs 52.4 +/- 12.6 yrs, respectively; p < 0.005). The cumulative MTX dose ranged from 65 to 580 mg at the time pneumonitis developed. Five of the patients (83%) had preexisting interstitial abnormalities, while only 29 of the 278 patients without MTX pneumonitis (10%) had such abnormalities (p < 0.001). The frequency of adverse effects due to previous treatment with disease modifying antirheumatic drugs (DMARD) was 66.7% in MTX pneumonitis patients and 14.3% in the other 278 patients (p < 0.01). CONCLUSION: Advanced age, preexisting interstitial abnormalities, and previous adverse reactions to DMARD may be associated with MTX pneumonitis. Patients with these characteristics require careful monitoring during MTX therapy. PMID- 9415632 TI - Relationship between duration of rheumatoid arthritis before knee joint surgery and HLA-DRB1 alleles: a retrospective study. AB - OBJECTIVE: To examine whether genetically determined factors can be used as predictors of requirement for knee joint surgery in the early phase of rheumatoid arthritis (RA). METHODS: We determined HLA-DRB1 alleles in 322 patients with seropositive RA by polymerase chain reaction and allele specific oligonucleotide probe techniques. Patients were classified into 3 groups (S/S, S/N, and N/N) based on their possession of two, one or no susceptibility alleles of RA, respectively. The stage of radiographic change in the knee joint determined using Larsen's standard film was compared to results of genotyping. Duration of RA before joint surgery in the 3 groups was also compared retrospectively. RESULTS: The median number of years to develop to stages I, II, III, and IV and the number of years of disease duration before total knee arthroplasty (TKA) were significantly shorter in the S/S group than in the S/N and N/N groups (p < 0.05). CONCLUSION: TKA was required earlier in the S/S group than in the S/N and N/N groups. This finding will affect planning of surgical management for RA based on anticipated courses. PMID- 9415633 TI - Fluoride therapy in prevention of rheumatoid arthritis induced bone loss. AB - OBJECTIVE: To determine the efficacy of sodium fluoride (40 mg/day) in preventing rheumatoid arthritis (RA) induced bone loss, which may lead to osteoporosis. METHODS: We conducted an 18 month, randomized, double blind, placebo controlled trial in 38 patients with RA. The primary outcome measure was the difference in the percentage change between groups in lumbar spine bone mineral density (BMD) from baseline values after 18 months of therapy. The secondary outcome measures were the differences in the percentage change between groups in femoral neck, Ward's triangle, trochanter, and total body BMD from baseline after 18 months of therapy. RESULTS: There was a significant percentage difference (SD) between groups of 6.2% (7.3%) (p = 0.0005) in lumbar spine BMD after 18 months of treatment in favor of the fluoride group. The fluoride group experienced a 5.2% (8.4%) (p = 0.0125) increase, whereas the placebo group showed a 1.0% (4.8%) (p = 0.8015) decrease in lumbar spine BMD after treatment. No significant differences were found for the femoral neck, Ward's triangle, trochanter, and total body BMD in terms of the percentage changes from baseline within each treatment group or in the differences in the degree of change between groups after therapy. Lumbar spine BMD increased in about 80% of patients treated with fluoride (responders) compared to 44% of patients treated with placebo. CONCLUSION: The results showed that fluoride therapy was well tolerated and increased vertebral bone mass in patients with RA. PMID- 9415634 TI - Genetic variation in the interleukin 10 gene promoter and systemic lupus erythematosus. AB - OBJECTIVE: To investigate interleukin 10 (IL-10) gene promoter polymorphisms in systemic lupus erythematosus (SLE) and its clinical subsets. METHODS: DNA from 76 Caucasian patients with SLE and 119 controls as genotyped for 3 defined dimorphic polymorphisms (G or A at position -1082, C or T at position -819, C or A at position -592) in the promoter region of the IL-10 gene, using the polymerase chain reaction to amplify the IL-10 gene promoter and oligonucleotide probes specific for each allelic sequence. The frequency of genotypes was compared between patients with SLE and controls, and between clinical subsets of patients with the disease. RESULTS: There was no significant change in the allele frequency of the three IL-10 gene promoter dimorphic polymorphisms in the SLE group compared with controls. However, when subgrouped according to autoantibody status and clinical features, IL-10 -1082*G, -819*C, and -592*C alleles were increased in patients possessing Ro autoantibodies and those with renal involvement. These alleles are in preferential allelic association, namely GCC, ACC, and ATA haplotypes, and the GCC haplotype was increased in these patient subgroups. CONCLUSION: Polymorphisms within the IL-10 gene promoter that are associated with high IL-10 levels may be important in the development of certain clinical features in SLE. PMID- 9415635 TI - Clinically occult avascular necrosis of the hip in systemic lupus erythematosus. AB - OBJECTIVE: To study the natural history of clinically occult avascular necrosis (AVN) of the hip in patients with systemic lupus erythematosus (SLE). METHODS: Sixty-six patients with SLE (without symptoms referable to the hip) receiving at least 5 mg/day prednisone for > or = 6 months were screened by magnetic resonance imaging (MRI) for AVN of the hip. A complete MRI evaluating class and percentage of femoral head involvement, AP and lateral radiographs of the hips, bone scan, and physical examination were performed for patients with positive MRI. Medical records were reviewed for serologic and clinical variables that might predict AVN. Repeat MRI were obtained at 3, 6, and 12 months to assess possible progression or resolution of the lesion. Patients with negative screening MRI underwent repeat screening after one year to assess the one year incidence rate. RESULTS: Eleven asymptomatic hips (8%) in 8 patients (12%) had MRI documented AVN. The percentage of femoral head involvement ranged from 1 to 46%. One lesion was MRI class B, the remaining lesions were class A. The radiographic stage of 10 hips was stage 1, the MRI class B hip was stage 2. Risk factors for clinically occult AVN included Afro-American origin, Raynaud's phenomenon, migraine headaches, and a maximal corticosteroid dose of at least 30 mg/day. After 12 months, 43 of 58 patients with an initially negative MRI underwent repeat screening examinations; no new lesions were observed. CONCLUSION: Clinically occult AVN of the hip is common in patients with SLE. The short term natural history of these lesions appears stable without spontaneous healing or clinical or radiographic progression. Risk factors for these asymptomatic lesions are similar to the risks for symptomatic AVN and surgical intervention appears not to be indicated in these patients. PMID- 9415636 TI - Neurochemistry of brain lesions determined by spectroscopic imaging in systemic lupus erythematosus. AB - OBJECTIVE: The significance and etiology of focal brain lesions in systemic lupus erythematosus (SLE) are unknown. Our purpose was to determine whether the neurochemistry of focal lesions and normal appearing brain tissues in SLE were consistent with neuronal loss, demyelination, or ischemia. METHODS: Patients with SLE (n = 14) and controls (n = 13) were studied using magnetic resonance imaging (MRI) and spectroscopic imaging (SI) at 1.5 Tesla. RESULTS: MRI detected fixed focal brain lesions (n = 16) and SI measured brain metabolites, including N acetylaspartate (NAA), creatine (Cre), choline (Cho), and lactate (Lac). NAA/Cre of normal appearing brain was decreased in patients with SLE compared to controls: grey matter (1.74 +/- 0.16 vs 1.92 +/- 0.18; p = 0.01), occipital white matter (1.98 +/- 0.22 vs 2.23 +/- 0.16; p = 0.004), and periventricular white matter (2.00 +/- 0.23 vs 2.33 +/- 0.23; p = 0.001). Lesions were characterized by markedly decreased NAA/Cre relative to normal appearing tissues in the same patient (1.67 +/- 0.22 vs 1.88 +/- 0.14; p = 0.0002). Elevated Cho/Cre was observed in 25% of focal lesions and 21% of normal appearing tissues. No elevation of lactate was observed in lesions or normal appearing tissues. CONCLUSION: SI detects focal and generalized brain abnormalities in SLE characterized by decreased NAA, elevated choline, and normal lactate. These findings are consistent with widespread neuronal injury and demyelination, but are not consistent with anaerobic metabolism. PMID- 9415637 TI - Relationship between endocrine, immune, and clinical variables in patients with systemic lupus erythematosus. AB - OBJECTIVE: To assess the frequency of increased plasma prolactin (PRL) in patients with systemic lupus erythematosus (SLE) and to evaluate its relationship to other hormonal and immune variables. METHODS: Thirty-five patients with SLE with various levels of disease activity were studied. Plasma PRL, cortisol, growth hormone (GH) were determined by radioimmunoassay and interleukin 6 (IL-6) by ELISA: SLE activity was evaluated using the European Consensus Lupus Activity Measurement (ECLAM). RESULTS: Increased plasma PRL concentration (> 20 ng/ml) was recorded in 11 patients (31%). No correlation was found between plasma PRL and GH, IL-6, cortisol, or C-reactive protein, nor was any significant correlation observed between plasma PRL and the ECLAM score. Patients with hyperprolactinemia were, however, found to have been treated with higher doses of prednisone therapy than patients with normal plasma PRL. Further analysis of the relationship of plasma PRL and therapy showed that patients with SLE selected by the attending physician for prednisone therapy in doses > or = 10 mg/day were more frequently hyperprolactinemic. CONCLUSION: Our findings that patients with SLE with a more active form of the disease and who are less responsive to therapy had increased plasma PRL levels more frequently may be indicative of a potential relationship of hyperprolactinemia to severity of disease. PMID- 9415638 TI - Literacy in patients with a chronic disease: systemic lupus erythematosus and the reading level of patient education materials. AB - OBJECTIVE: (1) To assess literacy in a sample of patients with systemic lupus erythematosus (SLE); (2) to evaluate the reading level of patient education materials specific to SLE; and (3) to compare patient literacy levels to the readability of materials written for patients with SLE. METHODS: Rapid Estimate of Adult Literacy in Medicine, a reading recognition test, was given to 94 patients with SLE. Socioeconomic status was assessed using Nam-Powers. Patient education materials frequently used with these patients were assessed for readability grade level. RESULTS: The patients with SLE were reading on an average 7th-8th grade level; their average educational level (last grade completed in school) was 11.9. The average socioeconomic status (SES) according to the Nam-Powers assessment was 43, indicating high school completed, no college, an income range of $5000-$10,000, and occupations such as household workers and laborers. Multiple linear regression revealed that race and education correlated with reading (p < 0.001), but age, sex, and SES did not. The readability of surveyed SLE patient education materials ranged from 7th-15th grade level. Eighty-nine percent were written at a 9th grade level or above and were therefore inappropriate for about half the patients surveyed. CONCLUSION: Reading skills below high school level existed for 48% of patients surveyed, yet only 11% of SLE patient education materials were written below a 9th grade level. Current SLE patient education materials are written on too high a level for many patients. Identifying patients with low literacy may help provide more appropriate patient education and better medical care. PMID- 9415639 TI - T cell reactivity to Sjogren's syndrome related antigen La(SSB). AB - OBJECTIVE: Many patients with primary Sjogren's syndrome (SS) make high titer IgG autoantibodies to the La(SSB) antigen, suggesting antigen specific T cell-B cell interactions. T cell responses to some nuclear antigens, particularly U1RNP, have been detected in patients with systemic lupus erythematosus (SLE) and in healthy subjects. We investigated T cell reactivity to the autoantigen SSB in patients with SS and healthy controls. METHODS: Using the [3H]thymidine proliferation assay, we determined reactivity to purified recombinant SSB (rSSB) in 20 patients with SS and 19 controls. Specificity was determined using tetanus toxoid, endotoxin, and 3 other autoantigens (PBC.M2, Sc170, and GAD). Precursor frequency was calculated by limiting dilution analysis. HLA Class II dependency was investigated using anti-Class II monoclonal antibodies. HLA-DR typing was by polymerase chain reaction and sequence specific oligonucleotide typing. RESULTS: Six of 20 patients with SS and 10/19 controls proliferated to La(rSSB). Precursor frequency of anti-SSB T cells was 1:77,040 and 1:115,000 in 2 healthy subjects and 1:230,250 and 1:103,034 in two patients with SS. Anti-HLA-DR abrogated proliferation to SSB and tetanus toxoid. Thirteen of 15 patients with SS and 4/17 controls were HLA-DR3 positive, with no apparent association of HLA-DR3 with SSB reactivity in controls. CONCLUSION: Anti-La(SSB) specific T cells occur in a significant proportion of controls and in some patients with SS. The function of SSB T cells in controls remains to be defined. They may represent immunoregulatory cells, and further analysis of these cells, and a comparison to those found in patients with SS, may elucidate normal immunoregulation and the derangements that lead to Sjogren's syndrome. PMID- 9415640 TI - Geographical clustering of mortality from systemic sclerosis in the Southeastern United States, 1981-90. AB - OBJECTIVE: To determine whether elevated rates of mortality from systemic sclerosis (SSc) in the Southeastern United States result from local, multicounty clusters of the disease. METHODS: Detection of spatial clusters of SSc mortality by applying the method of Kulldorff and Nagarwalla to death certificate data from 955 counties in 12 southeastern states. RESULTS: From 1981 to 1990, significant excess mortality from SSc in the Southeastern US occurred among white males [standardized mortality ratio (SMR) = 1.2; p = 0.0004] and black males (SMR = 1.2; p = 0.04), but not among white females (SMR = 0.98; p = 0.55) or black females (SMR = 1.1; p = 0.06). When the cluster detection algorithm was applied to data for white males, 3 significant clusters were identified. The primary cluster (p = 0.001) was centered around Coffee, Tennessee. Two smaller clusters overlapped the primary cluster -- one centered at Calhoun, Alabama, (p = 0.008) and another centered at Chattooga, Georgia, (p = 0.04). Analysis of data for black males resulted in a single significant cluster (p = 0.02) centered at Northampton, North Carolina. When data for white or black females were analyzed, no clusters reached statistical significance. In combination, excess SSc mortality in the detected clusters accounted for 79.0 and 66.2%, respectively, of the excess deaths among white and black males across the whole Southeast. CONCLUSION: Elevation of SSc mortality rates in the Southeastern US results from local clusters of concentrated mortality. These clusters may be artifacts of regional variation in death certificate quality. If not, distinctive environmental factors in these areas may provide new insights into the etiology of SSc. PMID- 9415641 TI - Ascorbic acid absorption in patients with systemic sclerosis. AB - OBJECTIVE: To investigate whether reduced circulating levels of ascorbic acid in patients with systemic sclerosis (SSc) are a result of malabsorption. METHODS: Eight patients with SSc, but with no evidence of bacterial overgrowth, and 8 healthy controls were recruited. On the first day of study, each subject was given orally an aliquot of [14C] ascorbic acid, which was then "flushed out" by oral intake of unlabeled ascorbic acid for the following 7 days. Plasma samples were collected at specified intervals and urine was collected continuously over the 8 day study period. [14C] content of plasma and urine were measured by scintillation counting. For each subject, a plasma [14C] decay curve was drawn. Each subject's ascorbic acid absorption was assessed using the area under the curve (AUC) and the apparent renal clearance (CLr[app]). Ascorbic acid intake was assessed using dietary history and food composition tables. RESULTS: There were no differences in the dietary intake of vitamin C (p = 0.16) and body mass indices (p = 0.91) between patients and controls. The plasma [14C] AUC and CLr(app) were similar between patients and controls [AUC patient mean (standard deviation, SD) = 37.1 (6.8), AUC control mean (SD) = 38.6 (9.9), p = 0.74; CLr(app) patient mean (SD) = 0.57 (0.24), CLr(app) control mean (SD) = 0.47 (0.27), p = 0.45]. CONCLUSION: There was no evidence of impaired absorption of ascorbic acid in patients with SSc without bacterial overgrowth compared to healthy controls. PMID- 9415642 TI - Noninvasive assessment of myocardial involvement in patients with systemic sclerosis: role of signal averaged electrocardiography. AB - OBJECTIVE: To assess the role of late ventricular potentials (LVP) in detecting early myocardial involvement in patients with systemic sclerosis (SSc). METHODS: Seventy-seven patients with SSc (68 women, 9 men, mean age 50 +/- 13 yrs) and 33 control subjects (18 women, 15 men, mean age 46 +/- 10 yrs) underwent resting electrocardiogram (ECG), 24 h Holter monitoring, complete echocardiographic and Doppler echocardiographic examination, and signal averaged ECG at high pass setting of 40 Hz, with the low pass fixed at 250 Hz. Patients with SSc underwent resting myocardial scintigraphy and radionuclide angioventriculography. RESULTS: The prevalence of LVP at 40 Hz was 20.5%. Compared to control subjects, patients with SSc showed higher prevalence of septal infarction pattern (p = 0.05), complex ventricular arrhythmias (p = 0.03), pulmonary arterial hypertension (p < 0.001), and LVP (p = 0.02). Forty-four patients with SSc (57.1%) had resting perfusion defects by myocardial scintigraphy. Fourteen of 15 patients with LVP showed perfusion defects compared to 29 of 58 without LVP (p = 0.002). Linear regression analysis showed that myocardial perfusion defect score was significantly correlated to either the filtered QRS duration, or the duration of low amplitude signals < 40 microV of the terminal QRS, or the root mean square voltage of the last 40 ms of the QRS complex. After a mean followup period of 20 months, 8 patients died. In 2 patients who died suddenly, LVP were present. CONCLUSION: Signal averaged ECG is a sensitive and inexpensive technique in the clinical assessment and followup of patients with SSc. PMID- 9415643 TI - Enhanced monocyte generation of reactive oxygen species in primary systemic vasculitis. AB - OBJECTIVE: Little is known about the role of mononuclear phagocytes in systemic vasculitis. We examined the hypothesis that monocytes from patients with primary systemic vasculitis have increased capacity to generate reactive oxygen species. METHODS: We studied patients with primary systemic vasculitis (n = 24) and compared them with patients with autoimmune related vasculitis (20), peripheral vascular disease (12), and postrenal transplantation (8). Healthy controls were also studied (56). Peripheral blood monocytes were stimulated using phorbol myristate acetate (PMA). We measured the generation of superoxide by cytochrome c reduction and myeloperoxidase (MPO) dependent products using peak luminol chemiluminescence. RESULTS: Patients with primary systemic vasculitis generated significantly more superoxide than all 3 non-vasculitis groups. Superoxide generation in the autoimmune vasculitis group was also higher than other groups but failed to reach significance versus controls (p = 0.07). Generation of MPO dependent products was also significantly higher in patients with primary vasculitis compared to all other groups. Subgroup analysis within the primary vasculitis group showed no correlation of these findings with disease activity, antineutrophil cytoplasmic antibodies, or American College of Rheumatology classification. CONCLUSION: Mononuclear phagocytes act as a source of reactive oxygen species in primary systemic vasculitis. Further study is required to determine if the reactive oxygen species generated act as a mechanism to promote vessel wall injury, or healing in this condition. PMID- 9415644 TI - Raynaud's phenomenon and digital gangrene as a consequence of treatment for Kaposi's sarcoma. AB - OBJECTIVE: To analyze the clinical features and to identify factors associated with the development of Raynaud's phenomenon (RP) in patients receiving chemotherapy for human immunodeficiency virus (HIV) related Kaposi's sarcoma. METHODS: A retrospective cohort study of all patients with Kaposi's sarcoma treated with chemotherapy at Toronto-Sunnybrook Regional Cancer Centre from 1987 to 1995. Patients who developed RP were compared with those who did not with respect to age, CD4 cell count, Acquired Immune Deficiency Syndrome Clinical Trials Group (ACTG) stage, smoking history, type and dose of chemotherapy, and previous treatment with interferon and radiation therapy. RESULTS: Eighty-seven patients with Kaposi's sarcoma were treated with chemotherapy between 1987 and 1995. Five developed RP, which progressed to digital gangrene. Median age, proportion of smokers, proportion defined as ACTG poor risk, median CD4 count, and history of opportunistic infections were equal in the 2 groups. All patients with RP received vinblastine followed by bleomycin. No cases of RP occurred in 27 patients treated with vinblastine alone or in 24 patients treated with bleomycin without previous vinblastine. Patients developing RP tended to have received higher cumulative doses of chemotherapy including bleomycin (p = 0.067) and previous treatment with either local radiation or interferon (p < 0.009, p < 0.001, respectively). CONCLUSION: Sequential chemotherapy with vinblastine followed by bleomycin was associated with the development of RP in patients with HIV related Kaposi's sarcoma. Bleomycin alone was not associated with RP. PMID- 9415645 TI - Differential risk of non-Hodgkin's lymphoma in Italian patients with primary Sjogren's syndrome. AB - OBJECTIVE: To assess the incidence of non-Hodgkin's lymphoma (NHL) and estimate the relative risk (RR) of developing lymphoproliferative complications in a large population of Italian patients with Sjogren's syndrome (SS) and to ascertain if any difference exists between the north and centre-south of Italy. METHODS: Differential relative risks of NHL were obtained by comparing the number of observed cases with cases identified on the basis of age-sex-time specific incidence rates extracted from regional cancer registries. RESULTS: Among the 331 patients with SS studied, 9 cases of NHL occurred, while no lymphoid malignancy appeared in patients with overlapping connective tissue disease (secondary SS) or in males with primary SS. As the number of NHL cases identified on the basis of the rate in the cancer registries would have been 0.27, the RR is 33.3 (p < 0.001). The incidence rate of NHL in Italian patients with SS is about 5.1/1000 person-years. 5.4/1000 per year in the north of the country and 4.8/1000 per year in the centre-south. The relative risks are, respectively, 34.7 and 32.5. CONCLUSION: Italian patients with primary SS have increased risk of developing NHL. In this group, the absence of a significant difference between the north and the centre-south of Italy contrasts with the higher incidence of NHL in the general population of northern regions and strengthens the direct connection between primary SS and NHL. PMID- 9415646 TI - Which patients with ankylosing spondylitis derive most benefit from an inpatient management program? AB - OBJECTIVE: To evaluate the benefit achieved from an inpatient management program by testing the hypothesis that more mobile, younger patients on their first course show the most improvement. METHODS: We assessed 236 patients over an 18 month period. Patients were assessed at the beginning and end of the course by 4 indices, 3 of which were self-administered (disease activity, functional ability, and global well being) and one administered by a trained physiotherapist (metrology). Paired t tests and ordinary least squares regression analysis were used to compare pre and postcourse results for each index. RESULTS: The wide range of disease duration (0-53 years) and disease severity [0.69-9.39 (on a 0-10 scale)] reveal that patients are from a wide spectrum of disease. Results revealed a mean improvement of 18-27% over baseline in the 4 indices. Significant predictors of greater improvement over the 2 week course were found for each index. CONCLUSION: Our results confirm the benefit of an intensive regime of inpatient physiotherapy. Although the original hypothesis cannot be accepted or rejected as the results were different for the 4 indices, the following general conclusions may be drawn: (1) there is a trend for females to improve more than males; (2) patients who have attended fewer courses tend to achieve more improvement; (3) younger patients do better than older patients; and (4) age of disease onset has little effect. PMID- 9415647 TI - Monosodium urate, hydroxyapatite, and calcium pyrophosphate crystals induce tumor necrosis factor-alpha expression in a mononuclear cell line. AB - OBJECTIVE: Tumor necrosis factor-alpha (TNF-alpha) is thought to be important in chronic inflammation of joints characteristic of crystal induced arthritis. Monocytes are instrumental in maintaining that inflammation. We investigated production of mRNA and protein for TNF-alpha in vitro in a murine mononuclear cell line, after exposure to relevant crystal types. METHODS: Using the cell line designated RAW 264.7, cells were grown in standard medium and exposed to varying amounts of monosodium urate (MSU), hydroxyapatite (HA), and calcium pyrophosphate dihydrate (CPPD) crystals for differing time periods. Analysis of TNF-alpha mRNA induced by such exposure was by Northern hybridization; analysis of TNF-alpha protein was by ELISA: RESULTS: RNA analyses of cells treated with various levels of MSU, HA, and CPPD crystals showed strong induction of TNF-alpha transcripts. ELISA on culture supernatants confirmed high level TNF-alpha peptide secretion resulting from that transcriptional upregulation. Time course studies showed peak accumulation of TNF-alpha mRNA 1-6 h post-treatment. Study of the signalling pathway involved in TNF-alpha transcriptional upregulation indicated that increased phospholipase A2 activity was required. CONCLUSION: These observations suggest that crystals in joints can directly stimulate production of TNF-alpha, and that the source of that cytokine may be the monocytes known to be present and playing an important role in chronic joint disease. PMID- 9415648 TI - Acute gouty synovitis associated with "urate milk". AB - OBJECTIVE: To analyze the clinical features of acute gouty synovitis associated with thick, milky white, "chalky," urate laden synovial effusions, and to investigate the effects on synovial white blood cell (WBC) counts when leukocyte rich rheumatoid effusions are incubated with a urate packed milky synovial fluid. METHODS: Five patients (all men, mean age 70.8 years) with acute gouty synovitis (acute arthritis in 3, acute bursitis in 2) associated with "urate milk" were studied between 1993 and 1996. RESULTS: Synovial effusions were thick, "chalky," and appeared "milky" white. The fluids were packed with monosodium urate (MSU) crystals, which sedimented upon standing, leaving a clear supernatant containing a few MSU crystals. The presence of massive amounts of MSU crystals and crystal clumps interfered with accurate determination of synovial WBC counts. Four fluids showed "a few leukocytes," and one a WBC count of 6750/mm3 with 91% neutrophils and several intraleukocytic crystals. Four patients had subcutaneous tophi. Of the risk factors associated with development of gout, the most frequent was ethanol abuse, in 4 and possibly all 5 patients. Incubation of leukocyte-rich rheumatoid synovial effusions with urate laden knee fluid from Patient 5 produced a greater reduction in synovial WBC counts compared to controls. CONCLUSION: Milky white synovial effusions containing massive quantities of urate crystals (referred to as "urate milk") may rarely occur in the setting of acute gouty arthritis or bursitis. Ethanol abuse appears to be a risk factor associated with the development of hyperuricemia and gout in these patients. PMID- 9415649 TI - Leukemia inhibitory factor (LIF) binding protein attenuates the phlogistic and abolishes the chondral effects of LIF in goat joints. AB - OBJECTIVE: To investigate the ability of murine leukemia inhibitory factor (LIF) binding protein (mLBP) to attenuate the effects of recombinant human LIF (rhLIF) in goat radiocarpal joints in vivo. METHODS: Endotoxin-free saline (1 ml) containing either 0.5 or 1 microg of rhLIF was injected into the left and right radiocarpal joints of male angora goats. One hour later the right radiocarpal joints were injected with either 1 or 5 microg of naturally occurring mLBP in 1 ml saline, while the left radiocarpal joints (controls) received 1 ml saline vehicle alone. Goat joints were examined for clinical features of inflammation and synovial fluid (SF) was aspirated on Day 0 (before injection) and Days 2 and 6 postinjection. Leukocyte counts and concentrations of keratan sulfate were determined in the SF. Proteoglycan synthesis and proteoglycan content of cartilage was determined ex vivo in cartilage explants obtained at Day 6. RESULTS: Preliminary time course studies in vitro showed that mLBP had to be added to cartilage explant cultures within 1 h of rhLIF for effective antagonism to occur. In joints injected with either 0.5 or 1 microg rhLIF significant increases in swelling, effusion volume, leukocyte counts, and SF keratan sulfate concentrations were observed relative to controls. Statistically significant depressions of ex vivo proteoglycan synthesis and in proteoglycan content of articular cartilage were also observed relative to controls. In joints injected with 1 microg rhLIF followed by 1 microg mLBP, statistically significant improvement was only observed in the rate of ex vivo cartilage proteoglycan synthesis. The observed rate did not differ significantly from that obtained in joints treated with vehicle alone. In contrast, in joints injected with 0.5 microg rhLIF followed by 5 microg mLBP, statistically significant improvement was observed in all variables. Treatment with 5 microg mLBP effectively negated the effects of rhLIF on joint swelling, effusion volume, leukocyte infiltration, and cartilage proteoglycan catabolism. CONCLUSION: Murine LBP has the ability to attenuate the phlogistic effects of rhLIF in radiocarpal joints of goats and also abolishes the stimulatory effect of rhLIF on cartilage proteoglycan catabolism and depression of ex vivo proteoglycan synthesis. These antiinflammatory and chondral effects suggest that a humanized derivative of mLBP could be a clinically useful antagonist for LIF in inflammatory diseases. PMID- 9415650 TI - The relationship between variations in knee replacement utilization rates and the reported prevalence of arthritis in Ontario, Canada. AB - OBJECTIVE: To determine the relationship between regional variations in knee replacement (KR) utilization rates in Ontario, Canada, and the reported prevalence of arthritis and rheumatism as a chronic health problem. METHODS: Utilization data were acquired from the Canadian Institute for Health Information for KR procedures performed in Ontario between fiscal years 1984 and 1990. Census information was obtained from Statistics Canada. Disease prevalence data were derived from the 1990 Ontario Health Survey (OHS). Public Health Units (PHU) were used as the unit of analysis, with utilization rates defined as the number of KR performed on all PHU residents (irrespective of where these procedures were performed) divided by the population. Direct methods were used to standardize utilization for age, sex, and disease prevalence. The extremal quotient, the weighted coefficient of variation, and the systematic component of variation were used as measures of variation. The relationship between the number of KR performed in each age-sex-year strata and various demographic (age and sex), disease prevalence, and regional dummy variables was estimated using a Poisson regression model. RESULTS: Regional variation in the standardized utilization of KR surgery was wide, but declined over the study period; the extremal quotient fell from 8.0 to 3.3, the weighted coefficient of variation fell from 0.49 to 0.30, and the systematic component of variation fell from 0.20 to 0.17. Variation in the provision of KR surgery remained even after controlling for the demographic composition of the population and disease prevalence. Moreover, while demographic, regional, and temporal covariates were significant (p < 0.0001) in accounting for over 90% of the variation in utilization, disease prevalence was not significant (p > 0.05). CONCLUSION: This study merged population based reports of disease prevalence with administrative data to account for regional variations in utilization. While regional variations in KR surgery have fallen over time, variations remain even after adjusting for patient reported disease prevalence. The finding that demographic variables and the reported prevalence of disease were poorly correlated suggests that current area variation studies may not be adjusting fully for disease prevalence or severity. PMID- 9415651 TI - Validation study of a computerized version of the Western Ontario and McMaster Universities VA3.0 Osteoarthritis Index. AB - OBJECTIVE: To study the validity and feasibility of a computerized version of the Western Ontario and McMaster Universities (WOMAC) Osteoarthritis Index. METHODS: Thirty patients with osteoarthritis (OA) of the knee completed both a paper and a computerized version of WOMAC in random order. The visual analog scaled version of WOMAC, VA3.0, was used. We studied criterion validity by comparing the paper and computerized versions. RESULTS: All patients completed the computerized version without undue difficulty. Criterion validity, based on aggregated subscale scores, was excellent: Pain, ICC = 0.89, Stiffness, ICC = 0.87, Physical Function, ICC = 0.95. CONCLUSION: The computerized version of WOMAC VA3.0 is a valid alternative to the paper version. PMID- 9415652 TI - Differences in health status of older adults with pain in the hip or knee only and with additional mobility restricting conditions. AB - OBJECTIVE: To determine differences in health status of people aged 55 to 74 years with pain in the hip or knee only and with additional mobility restricting conditions. METHODS: A subsample from a community based study on pain, disability, comorbidity, and radiological osteoarthritis (OA) was used to identify a group with current pain in the hip or knee only (n = 62), a group with additional mobility restricting conditions (n = 124), and a reference group without pain and radiological OA (n = 72). Health status was measured with the IRGL instrument (Impact of Rheumatic diseases on General health and Lifestyle). Additional mobility restricting conditions were self-reported. RESULTS: The most reported additional conditions were more widespread joint pain and stiffness, and cardiovascular and respiratory problems. The group with pain in the hip or knee only had less mobility than the reference group (p < 0.05), but had higher mobility (p < 0.05), less pain (p < 0.001), less psychological distress (p < 0.01), and less effect of symptoms on daily life (p < 0.001) than the group with additional conditions. No differences were found in background variables or comorbidity. Multivariate logistic regression analysis showed that the group with additional conditions differed from the group with knee or hip pain only with respect to joint pain (OR 1.18), cheerfulness (OR 0.9), and effect on daily life (OR 1.1). CONCLUSION: The health status of people with pain in the hip or knee only is comparable to that of a reference group without pain. Health status is lower when pain in the hip or knee is present in combination with additional mobility restricting conditions. This last group is at greater risk of psychological distress and physical dysfunctioning. PMID- 9415653 TI - Patients with neck and shoulder complaints and multisite musculoskeletal symptoms -a prospective study. AB - OBJECTIVE: To examine reasons for and extent of health services utilization in patients seeking care for neck and shoulder pain in a population based primary care setting. METHODS: Patients seeking care for neck and shoulder pain were identified from medical records of 6526 patients visiting 6 primary care centers during a 2 week period. The extent of and reasons for health care utilization over the subsequent 12 month period were examined. RESULTS: Of 440 patients who consulted primary health care physicians for neck and shoulder pain, one-half had one or more additional episodes of care due to musculoskeletal (MSK) pain over the subsequent 12 months. One-quarter had additional episodes of care for pain in other sites than in the neck and shoulder. The total number of visits was twice the annual average for patients visiting the health centers, MSK symptoms accounting for half the visits. Twenty percent of the women and 7% of the men visited primary care physicians 10 times or more per year. CONCLUSION: The pattern of reasons for visits for MSK pain suggests that in about one-quarter of patients visiting primary care physicians for neck and shoulder pain, the local symptomatology is part of multisite MSK symptoms, resulting in frequent utilization of health services. PMID- 9415654 TI - Induction of cyclooxygenase-2 by parathyroid hormone in human osteoblasts in culture. AB - OBJECTIVE: Parathyroid hormone (PTH) induced bone resorption by osteoclasts depends on the presence of osteoblasts. PTH induced production of prostaglandins by osteoblasts and induction of bone resorption by prostaglandins suggest that these autacoids may be implicated in the effects of PTH on bone. Our objective was to determine if the increase in prostaglandin production induced in human osteoblasts by PTH is due to an increase in cyclooxygenase-2 (COX-2) expression. METHODS: Primary cultures of human osteoblasts were obtained from specimens of trabecular bone. Confluent cells were treated with PTH, dexamethasone or compound NS-398, a specific COX-2 inhibitor. The concentration of prostaglandin E2 (PGE2) in the supernatants was determined by radioimmunoassay and COX-2 mRNA levels evaluated by Northern blot. RESULTS: PTH induced COX-2 mRNA expression and PGE2 production. These effects were time and concentration dependent and were inhibited by dexamethasone. Compound NS-398 reduced PGE2 production to the same extent as dexamethasone, and neither compound had an additive effect on this variable. CONCLUSION: These results show that PTH induces COX-2 expression in human osteoblasts in culture and suggest that this isoenzyme is the main factor in the control of prostaglandin synthesis in these experimental conditions. PMID- 9415655 TI - Preliminary evidence for cyclosporin A as an alternative in the treatment of recalcitrant juvenile rheumatoid arthritis and juvenile dermatomyositis. AB - OBJECTIVE: To evaluate the safety and efficacy of cyclosporin A (CyA) with and without methotrexate (MTX) in refractory juvenile rheumatoid arthritis (JRA) and juvenile dermatomyositis (JDMS). METHODS: Twenty-two patients (17 with JRA, 5 with JDMS) with refractory disease were studied retrospectively. All received CyA at a mean dose of 3.2 mg/kg/day over a mean period of 16 mo (range 6-42). All other medications except nonsteroidal antiinflammatory drugs, prednisone, and hydroxychloroquine were discontinued. In addition, 16/22 patients received concomitant MTX. RESULTS: Improvements in laboratory variables, joint counts, joint swelling, and morning stiffness were observed in most of the children with JRA. Muscle strength increased and muscle enzyme levels decreased in the patients with JDMS. CyA treatment permitted prednisone to be discontinued in 5/20 and reduced by greater than 50% in 10/20 patients. There was no evidence of hepatic or bone marrow toxicity or lymphoproliferative disease. Serum creatinine increased in 13/22 patients, but the actual values all remained within normal limits. CONCLUSION: CyA may be an effective agent in the treatment of refractory JRA and JDMS and concomitant MTX seems to be well tolerated. These preliminary data also suggest that combined CyA/MTX therapy may be associated with further improvement in clinical outcome. PMID- 9415656 TI - Azathioprine in patients with juvenile chronic arthritis: a longterm followup study. AB - OBJECTIVE: To evaluate drug survival, efficacy, side effects, and longterm toxicity of azathioprine treatment in patients with juvenile chronic arthritis (JCA). METHODS: In an uncontrolled, prospective study we evaluated 129 consecutive patients with JCA refractory to therapy in whom azathioprine treatment was begun during 1980-1989. In the first 29 patients, a 2 year trial was planned, while for the remaining 100 patients the protocol was to continue until remission or dropout. The median treatment period was 13 months (range 3 days-8.5 yrs). Patients were assessed every 2 months for 2 years for efficacy, side effects, growth and need for glucocorticoids, and outcome evaluated in late 1996. RESULTS: Remission without drugs was attained in 19 patients (15%); in addition, temporary remission in patients continuing treatment was attained in 18 cases (14%). Treatment was discontinued due to side effects in 18 cases (14%); in two-thirds of these cases side effects occurred during the first 2 months. Of the total number of patients, 49 (38%) completed 2 years of treatment, with significant improvement in both clinical and laboratory indices of disease activity. Treatment had no noticeable effect on iridocyclitis. One patient died of cytomegalovirus infection during azathioprine treatment. CONCLUSION: Azathioprine is a useful drug in severe JCA, with a sustained effect and acceptable side effects. Even in cases of incomplete remission, its glucocorticoid sparing effect was noteworthy. PMID- 9415657 TI - Induction of invasive and degradative phenotype in normal synovial fibroblasts exposed to synovial fluid from patients with juvenile rheumatoid arthritis: role of mononuclear cell population. AB - OBJECTIVE: To investigate the effect of synovial fluid (SF) from patients with juvenile rheumatoid arthritis (JRA) on proliferation and induction of degradative and invasive phenotype in normal synovial fibroblasts, and to elucidate the contribution of SF cells to this activity. METHODS: SF and/or conditioned medium (CM) from SF cells were evaluated for their ability to (1) stimulate a proliferative response, (2) induce the "activated phenotype" capable of invading cartilage matrix, and (3) promote the release of key matrix metalloproteinases (MMP) in normal synovial fibroblasts. RESULTS: Proliferation of normal synovial fibroblasts exposed to SF or CM from SF cells of patients with JRA was up to 3 times greater than untreated controls. Concomitant with induction of an activated phenotype in the treated synovial fibroblasts, the activated form exhibited up to 250% invasiveness of cartilage matrix compared to untreated synovial fibroblasts (100%), in addition to releasing increased MMP activity, not normally associated with these quiescent cells. This induction was not solely due to tumor necrosis factor-alpha, transforming growth factor-beta, interleukin 1beta (IL-1beta), and IL-6, as SF and/or CM depleted of these cytokines sustained about 40% of their invasive and inducing ability. We observed that the mononuclear cell (MNC) population that infiltrated into the joint cavity secretes this "inducing activity," which can be maintained in culture up to several weeks. CONCLUSION: Our data suggest that the cellular component of SF releases soluble factor(s) that directly or indirectly contribute to (a) proliferation of synovial fibroblasts, and (b) production and release of extracellular MMP by synovial fibroblasts, thereby inducing a degradative and invasive phenotype culminating in cartilage and bone destruction. PMID- 9415658 TI - Childhood central nervous system lupus; longitudinal assessment using single photon emission computed tomography. AB - OBJECTIVE: Neuropsychiatric manifestations in children with systemic lupus erythematosus (SLE) occur in 30-60% of patients during the course of disease. Unlike other manifestations of childhood SLE, few laboratory studies and imaging modalities aid in the diagnosis of central nervous system (CNS) lupus. We and others have reported the usefulness of single photon emission computed tomography (SPECT) in the initial assessment of cerebral blood flow in children with active CNS involvement. We extend these observations to longterm followup using the SPECT scan to determine its usefulness in the subsequent course of CNS lupus in children. METHODS: Eleven children who developed CNS disease and fulfilled the classification criteria for SLE were included in an open pilot study. The patients were followed up to 3.5 years and presented with CNS manifestations: encephalopathy with or without grand mal seizures (N = 4), focal seizures with depression or hallucinations (N = 3), optic neuritis with transverse myelitis (N = 2), and psychosis with audiovisual hallucinations (N = 2). Initially, all children had lumbar puncture, SPECT, and serologic testing; 9 children had electroencephalogram (EEG), 7 had computerized tomography (CT), and 10 had magnetic resonance imaging (MRI). SPECT was repeated in 7 patients 1-4 months after the initial CNS event and thereafter in 10 patients annually. RESULTS: At the time of the initial CNS event, 9/11 children (82%) had normal results for lumbar puncture, 7/9 (78%) for EEG, 5/7 (71%) for CT, and 6/10 (60%) for MRI. All patients (100%) had diffusely abnormal SPECT: In addition, 5/11 (45%) tested positive for IgG antibodies to cardiolipin and dsDNA, and 4/11 (36%) had antibodies to Sm. In 5/7 children whose SPECT was repeated 1 to 4 months after the CNS event, additional perfusion defects were documented compared with initial SPECT: During the subsequent 1-3.5 years and concomitant with treatment, CNS manifestations resolved clinically, but none of the SPECT scans became normal. Perfusion defects improved over time in 4 patients and worsened in 6. CONCLUSION: SPECT scan remains a sensitive tool during initial CNS events in children with CNS lupus documenting the presence of damage during short term followup of 1-4 months. However, during longterm followup abnormalities documented by SPECT no longer correlate with the patient's clinical course, limiting the usefulness of SPECT as a clinical tool in children who recover from CNS disease. PMID- 9415659 TI - Remission of nonerosive polyarthritis associated with Sjogren's syndrome after autologous hematopoietic stem cell transplantation for lymphoma. AB - We describe a 50-year-old woman with seropositive rheumatoid arthritis and Sjogren's syndrome who underwent autologous blood stem cell transplantation after relapse of associated non-Hodgkin's lymphoma. This resulted in complete remission not only of the lymphoma, but also the arthritis. PMID- 9415660 TI - Recurrent acute scleroderma renal crisis complicated by thrombotic thrombocytopenic purpura. AB - Acute renal crisis as an early manifestation of scleroderma is underemphasized, and its recurrence after initial successful therapy is rare. We describe a 32 year-old woman who presented with scleroderma renal crisis. A second episode of apparent renal crisis, however, was complicated by thrombotic thrombocytopenic purpura, which led to pancreatitis, a large cerebral infarction, and fatal outcome despite intensive therapy. This case illustrates the complexity and severity of diffuse systemic sclerosis presenting with multiple, major organ complications. PMID- 9415661 TI - Cyclophosphamide induced water intoxication in a woman with Sjogren's syndrome. AB - Water intoxication is a well described complication of high dose intravenous (i.v.) cyclophosphamide therapy combined with forced hydration. Less well known is that water intoxication can develop even after low dose iv cyclophosphamide. To draw attention to this potentially life threatening complication, we describe a woman who developed acute water intoxication after treatment with low dose iv cyclophosphamide for a sensory neuropathy secondary to Sjogren's syndrome. Rheumatologists should be aware of this serious adverse effect of iv cyclophosphamide because this drug is being used increasingly for treatment of a variety of rheumatological diseases. The pathogenesis, clinical characteristics, treatment, and methods for prevention of cyclophosphamide induced water intoxication are discussed. PMID- 9415662 TI - Unilateral hemochromatosis arthropathy on a neurogenic basis. AB - We describe a 42-year-old man with right hemiplegia from cerebral palsy who presented with chronic left arm pain. Examination revealed 12 active joints, predominantly in the left hand. Radiographs showed characteristic changes of an advanced secondary osteoarthritic process in the left hand and only minimal changes on the right. Bone scan confirmed unilateral activity. Subsequent investigations diagnosed hemochromatosis. Unilateral arthropathy of hemochromatosis on a neurogenic basis has not been previously reported. PMID- 9415664 TI - The diagnostic conundrum of carpal tunnel syndrome. PMID- 9415665 TI - The diagnostic conundrum of carpal tunnel syndrome. PMID- 9415663 TI - Milwaukee shoulder with massive bilateral cysts: effective therapy for hydrops of the shoulder. AB - "Milwaukee shoulder" is often associated with large effusions that cannot be managed with conventional therapy. We describe a 75-year-old man whose massive hydrops of both shoulders was resistant to treatment with nonsteroidal antiinflammatory drugs (NSAID), multiple aspirations, and injections of corticosteroid. The effusions resolved completely after treatment with oral colchicine and an NSAID containing magnesium. PMID- 9415666 TI - The diagnostic conundrum of carpal tunnel syndrome. PMID- 9415667 TI - Herpes zoster encephalomyelitis in a patient with rheumatoid arthritis treated with low dose methotrexate. PMID- 9415668 TI - Peripheral neuropathy during interferon-alpha therapy in patients with cryoglobulinemia and hepatitis virus infection. PMID- 9415669 TI - Dupuytren's disease and diabetes mellitus. PMID- 9415670 TI - Coexisting SLE and HIV infection. PMID- 9415671 TI - Use of alternative medicine in a consecutive sample of patients with systemic lupus erythematosus. PMID- 9415673 TI - Prevalence and characteristics of rheumatic manifestations in patients infected with human immunodeficiency virus undergoing antiretroviral therapy. PMID- 9415674 TI - Adenosine plasma levels after low dose methotrexate administration. PMID- 9415672 TI - Aseptic meningitis as a side effect of intravenous immune gammaglobulin. PMID- 9415675 TI - Arthropathy of Wilson's disease presenting as noninflammatory polyarthritis. PMID- 9415676 TI - Heteropagus conjoined twins due to fusion of two embryos: report and review. AB - We report on a case of conjoined twinning (CT) consistent with fusion of two embryos followed by resorption of the cranial half of one of them, resulting in a normal male baby with the lower half of a male parasitic twin fused to his chest. Fluorescent in situ hybridization (FISH) studies suggested that the parasitic twin was male, and DNA typing studies demonstrated dizygosity. Although incomplete fission is the usual explanation for conjoined twins, the unusual perpendicular orientation of the parasite to the autosite supports a mechanism observed in mares in which early fusion of two embryos is followed by resorption due to compromised embryonic polarity. PMID- 9415677 TI - Congenital scoliosis (hemivertebra) associated with de novo balanced reciprocal translocation, 46,XX,t(13;17)(q34;p11.2). AB - We report on an 8-year-old girl with congenital scoliosis (segmented hemivertebra between the second and third lumbar vertebrae) and psychomotor developmental delay. She has a de novo reciprocal translocation, t(13;17)(q34;p11.2). Congenital scoliosis is one type of structural spine deformation and hemivertebra is the most common anomaly causing congenital scoliosis. The cause and the mode of inheritance of hemivertebrae are unknown. Our patient has a de novo balanced chromosome aberration and retains two copies of the LLGL gene, which is usually lacking in patients with Smith-Magenis syndrome (SMS). Since some SMS patients who showed a deletion at 17p11.2 had congenital scoliosis, it is likely that one (17p11.2) of the breakpoints in our patient is a candidate region for a hemivertebra locus. PMID- 9415678 TI - Variable expression of rib, pectus, and scapular anomalies with Robin-type cleft palate in a 5-generation family: a new syndrome? AB - We report on a 5-generation family with multiple musculoskeletal anomalies, including: Robin-type cleft palate, rib "dysplasia," scapular hypoplasia, and pectus excavatum. Robin-type clefts are known to be associated with various skeletal malformations; however, most of these include limb anomalies which are not present in this family. To our knowledge, there are no reports of similar conditions in the literature. The transmission through 5 generations and the presence of male-to-male transmission are consistent with autosomal-dominant transmission of a trait with variable expressivity. PMID- 9415679 TI - The return of thalidomide: are birth defects surveillance systems ready? AB - In the 1960s, thalidomide caused limb deficiencies in thousands of infants worldwide. The limb deficiencies were frequently of the intercalary type. As a result, numerous countries started birth defect surveillance programs. In 1967, the Centers for Disease Control (CDC) started the Metropolitan Atlanta Congenital Defects Program (MACDP), a population-based surveillance system, to provide early warning against new teratogens. Recent studies have shown that thalidomide may be beneficial for a range of conditions, including cancer and AIDS, and it may once again become widely available. Here, we examine the ability of MACDP to detect an increase in the birth prevalence of limb deficiency as an early warning of fetal exposure to thalidomide. We calculated base rates for all limb deficiencies, for bilateral nonsyndromic intercalary or preaxial deficiencies, and for all nonsyndromic intercalary limb deficiencies among Atlanta infants born from 1968 through 1993. We used relative risk estimates from previous studies and a range of pregnancy exposure rates for thalidomide. We tested the statistical power of MACDP to detect subtle changes in the birth prevalence of these defects using Poisson and cumulative sum (CUSUM) techniques. The base rates for all limb deficiencies, for bilateral intercalary or preaxial deficiencies, and for all intercalary limb deficiencies, were 0.53, 0.035, and 0.022/1,000, and the estimated relative risks were 175, 4,570, and 8,180, respectively. We varied the assumed rate of exposure to thalidomide from 1/10,000 to 5/100. With a 1/1,000 exposure rate, both Poisson and CUSUM techniques will detect a rate change in intercalary limb deficiency in about 6 months of monitoring, and a rate change in bilateral intercalary or preaxial deficiencies in about 12 months of monitoring. When monitoring all limb deficiencies, a pregnancy exposure rate of 3.5% or less would go unnoticed by the Poisson method and would take more than 50 years for the CUSUM method to signal an alarm with a 1/1,000 exposure rate. However, for rates of exposure less than 1/1,000, a progressively longer period of time or larger sample are needed to detect a rate change by both methods. Our findings highlight the importance of enlarging the monitored population and correct case classification in birth defects surveillance. PMID- 9415680 TI - Ventriculomegaly with radial and renal defects: prenatal diagnosis in two consecutive sibs. AB - We describe two consecutive mid-trimester fetuses of different sexes with identical anomalies of the upper limbs and the kidneys in association with severe ventriculomegaly. We compare this apparently autosomal recessive syndrome to VACTERL-H association, Fanconi anemia, and two other, so far unparalleled syndromes. Taking into account the absence of chromosome breaks, the associated changes of the amniotic fluid, and the renal histology, we conclude that we are dealing with a different entity. PMID- 9415681 TI - Ectodermal abnormalities in Kabuki syndrome. AB - We describe a girl with Niikawa-Kuroki (Kabuki) syndrome (NKS) with conical incisors, hypodontia, hypoplastic nails, and brittle hair. Abnormal teeth are common in NKS and support a hypothesis of autosomal dominant inheritance of the syndrome [Halal et al., 1989; Silengo et al., 1996]. Hair abnormalities have never been investigated in NKS. The ectodermal involvement in NKS could represent an important clue for the understanding of the pathogenesis of this syndrome. PMID- 9415682 TI - Tandem duplication of 11p12-p13 in a child with borderline development delay and eye abnormalities: dose effect of the PAX6 gene product? AB - We report on a girl with a duplication of chromosome band 11p12-->13, which includes the Wilms tumor gene (WT1) and the aniridia gene (PAX6). The girl had borderline developmental delay, mild facial anomalies, and eye abnormalities. Eye findings were also present in most of the 11 other published cases with partial trisomy 11p, including 11p12-->13. Recently, it was shown that introduction of additional copies of the PAX6 gene into mice caused very variable eye abnormalities. Therefore, a PAX6 gene dosage effect is likely to be present in mice and humans. The central nervous system may be less sensitive to an altered PAX6 gene dosage, which is consistent with the borderline developmental delay in the present patient. Urogenital abnormalities were absent in this patient and in most of the other patients with partial trisomy of 11p. Therefore, the effect of a WT1 gene duplication on the embryological development of the urogenital tract remains uncertain. PMID- 9415683 TI - Spherophakia associated with molybdenum cofactor deficiency. AB - Molybdenum cofactor deficiency is an autosomal recessive disorder characterized by lack of activity of the enzymes sulfite oxidase, aldehyde oxidase, and xanthine dehydrogenase or oxidase. The clinical manifestations are indistinguishable from those of isolated sulfite oxidase deficiency: craniofacial alterations, intractable neonatal convulsions, very severe mental retardation, lens dislocation, and death in the first decade of life. Lens dislocation is found in nearly all patients after neonatal age. In the present case it developed late (at the age of 8 years) and was preceded by bilateral spherophakia. We hypothesize that an abnormal relaxation of the zonular fibers is the cause of spherophakia in this disease; this causes lens dislocation eventually, after days, months, or years. PMID- 9415684 TI - Increase in fetal breech presentation in female carriers of Duchenne muscular dystrophy. AB - Female carriers of Duchenne muscular dystrophy (DMD) may demonstrate elevated serum creatine kinase (CK) and reduction of muscle dystrophin in all muscle types. We hypothesized that decreased dystrophin in uterine or pelvic girdle musculature might affect the obstetrical performance of females heterozygous for a dystrophin mutation. We reviewed the outcome of 34 deliveries resulting in 35 children from 13 women who were mothers of males attending a muscular dystrophy clinic. Obstetrical performance was examined retrospectively by chart review and patient contact. Of 35 children, 6 (17%) were delivered in the breech position, which is a fivefold increase above the national standards for term pregnancies. Of the six infants with breech presentation, two were males affected with DMD, one was a female heterozygote, one was a male who died perinatally, and the carrier status of the other two females is unknown. Most DMD affected males (12/14) were delivered in the vertex position. Thus, it is likely that maternal, rather than fetal, muscle weakness was the significant factor in determination of fetal position at term. We speculate that subtle changes in uterine or pelvic girdle muscle tone may contribute to a higher rate of fetal breech position in carriers of the DMD gene. PMID- 9415686 TI - Unusual autosomal recessive lymphatic anomalies in two unrelated Amish families. AB - We report on two unrelated Amish families with familial occurrence of unusual lymphatic anomalies. The first family had two children, a boy and a girl, with congenital chylothorax both of whom died as a consequence of this condition (one prenatally and one neonatally). The second family has two brothers with isolated cystic hygroma. Neither family has any other individuals affected with any type of lymphatic anomaly. Differential diagnosis and presumed autosomal recessive inheritance pattern will be discussed. Familial cystic hygroma not associated with hydrops fetalis and neonatal death has not been reported previously. PMID- 9415685 TI - Brachydactyly-short stature-hypertension (Bilginturan) syndrome: report on two families. AB - We report on two families with autosomal dominant brachydactyly of hands and feet and hypertension. All affected members of the first family had proportionate short stature. However, the propositus and the affected relatives in the second family were only short compared to unaffected relatives. The hypertension was medically responsive in all cases. The propositus in the second family had poor compliance and a striking generalized vasculopathy. All patients were of normal intelligence and had a normal facial appearance. The brachydactyly-short stature hypertension syndrome was first reported by Bilginturan et al. [1973] in a Turkish family and the families reported by us are Caucasian and Hispanic. The gene causing this condition in the original Turkish family was recently mapped to 12p. Our report expands our existing knowledge and the ethnic diversity of this syndrome. PMID- 9415687 TI - Case report of rec(7)dup(7q)inv(7)(p22q22) and a review of the recombinants resulting from parental pericentric inversions on any chromosomes. AB - We report a rare case of duplication for 7q22 --> 7qter and deletion for 7p22 --> 7pter, resulting from a meiotic recombination of a paternal pericentric inversion, inv(7)(p22q22). The newborn boy had the 7q trisomy syndrome. In addition, the diagnosis of chondrodysplasia punctata was made from lumbar and hand X-ray films taken soon after birth. Only two cases of rec(7)dup(7q), both in a single family, have been reported previously. We review 133 offspring with recombinations resulting from pericentric inversions on any chromosomes reported between 1981 and 1995. Of the 133 cases, 110 had a long-arm duplication and short arm deletion, while only 23 had a short-arm duplication and long-arm deletion. In 85 of the 133 cases, the mother was an inversion carrier (five carriers had two affected offspring), and in 46, the carrier was a father (one carrier had three affected offspring). Kaiser [Hum Genet 1984;68:1-47] reviewed 63 offspring with recombinations derived from a parental pericentric inversion reported between 1972 and 1981. In both surveys, recombinations resulting from pericentric inversions of chromosomes 1, 12, 19, and Y were not found. PMID- 9415688 TI - Patients' attitudes about autonomy and confidentiality in genetic testing for breast-ovarian cancer susceptibility. AB - The identification of BRCA1 and BRCA2, two breast-ovarian cancer susceptibility genes, has brought many ethical and social issues to the forefront. This paper presents the results of a survey assessing the attitudes of 238 unaffected first degree relatives of women with breast or ovarian cancer regarding the ethical issues of autonomy and confidentiality as they relate to BRCA1/2 testing. Baseline knowledge about BRCA1/2 and ethnic and psychosocial characteristics of our study population were examined to determine their association with women's attitudes. The majority of women (86-87%) felt that health care providers should not disclose the results of genetic tests for breast-ovarian cancer susceptibility to insurance companies or employers without written consent; however, only 56-57% felt that written consent should be required for a spouse or immediate family to receive this information. Ninety-eight percent of the women surveyed agreed that genetic testing for breast-ovarian cancer risk should be voluntary. Likewise, most women (95%) agreed that a person should be able to have genetic testing against a doctor's recommendation and 88% of the women surveyed agreed that parents should be able to consent to genetic susceptibility testing on behalf of their minor children. African American women were less concerned than Caucasian women about the protection of confidentiality in families, they were more likely to agree that an individual should still have access to testing when their physicians recommended against it, and they were more supportive of parents' rights to consent to genetic predisposition testing on behalf of their minor children. Women with coping styles characterized by higher optimism were more likely to favor access to genetic testing when a physician recommended against it, and to support parents' rights to consent to testing of their minor children. Therefore, the setting and manner in which genetic counseling and testing are delivered must be appropriately tailored to reflect these attitudinal differences and preferences. PMID- 9415689 TI - De novo mutations of the Patched gene in nevoid basal cell carcinoma syndrome help to define the clinical phenotype. AB - The demonstration that mutations in the Patched (PTCH) gene cause nevoid basal cell carcinoma syndrome (NBCCS) has led to the identification of the exact molecular lesion in a percentage of individuals with the syndrome. In addition, it has been possible to determine, through molecular analysis of parents and other relatives of these individuals, if the mutation is inherited or has arisen de novo. We have previously reported 28 mutations in individuals with NBCCS, and here we present an additional 4 novel mutations. We have also analyzed relatives of a number of the individuals in whom we have found mutations. In total we have identified 8 individuals who carry a de novo mutation in the PTCH gene. In 5 of these cases, clinical and radiological examination had not unequivocally ruled out a diagnosis in one of the parents. This helps to define the clinical phenotype and suggests that diagnostic criteria in this complex syndrome may require review. PMID- 9415690 TI - Prader-Willi and Angelman syndromes: diagnosis with a bisulfite-treated methylation-specific PCR method. AB - The putative promoter region of the SNRPN gene contains a CpG island which is heavily methylated in the maternally derived allele and unmethylated in the paternally derived allele. In patients with Prader-Willi syndrome (PWS) only the methylated allele is present, while in those with Angelman syndrome (AS) only the unmethylated allele is present. The purpose of this paper is to report a polymerase chain reaction (PCR)-based assay to evaluate methylation status of the CpG island of the SNRPN gene and to show that this assay allows rapid diagnosis of PWS and AS. Methylated cytosines in the CpG dinucleotide are resistant to chemical modification by sodium bisulfite. In contrast, bisulfite treatment converts all unmethylated cytosines to uracil. Based on this differential effect, the bisulfite-modified DNA sequence of a methylated allele was successfully distinguished from that of an unmethylated allele using 2 sets of allele-specific primer pairs: a methylated allele-specific primer pair (MET) and an unmethylated allele-specific primer pair (UNMET). Bisulfite-modified DNA from 10 patients with PWS amplified only with the MET pair while modified DNA from 5 patients with AS amplified only with the UNMET pair. Modified DNA from 50 normal unrelated individuals amplified with both primer pairs. In that methylation-specific PCR (MSPCR) can detect all presently testable causes of PWS and AS in a rapid and cost-effective fashion, serious consideration should be given to the use of this test in the initial evaluation of all patients in which PWS or AS is being considered. PMID- 9415691 TI - Educational material about genetic tests: does it provide key information for patients and practitioners? AB - Genetic testing for common conditions will be used increasingly in primary care, but resources for patient counseling are decreasing. It is also necessary that primary care practitioners be better equipped to do basic genetic counseling. Therefore, the quality of informational materials for practitioners and patients is important. It was unknown how often key elements recommended by policy groups were actually included in such material. It was our aim to determine the content of printed informational material for practitioners and patients on genetic testing. We performed (1) a telephone survey of organizations in the United States that developed genetic tests or services and (2) a content analysis of pamphlets obtained from these organizations to determine the presence of 10 critical elements necessary to evaluate the appropriateness and performance of the tests. Almost 95% (169/178) of organizations responded to our survey; 131/169 (78%) reported using informational materials. We analyzed 115 pamphlets collected from 125/131 (95%) organizations. Elements least frequently included in the pamphlets were risks and benefits, patient rights, and intended use or purpose of the test. Most frequently included were descriptions of the conditions detected by the test, and the appropriate patients for testing. Nearly one half of the pamphlets included some statement about the accuracy of the test, but most of these did not specify whether their statements referred to sensitivity, specificity, or predictive value. Overall, pamphlets tended to contain information that would aid in determining a patient's eligibility for a genetic test, but did not contain sufficient information about the tests themselves. Our results suggest that several critical elements need to be added to enhance informed choices by patients and physicians. PMID- 9415692 TI - XY sex reversal and gonadal dysgenesis due to 9p24 monosomy. AB - We describe a case of XY sex reversal, gonadal dysgenesis, and gonadoblastoma in a patient with a deletion of 9p24 due to a familial translocation. The rearranged chromosome 9 was inherited from the father; the patient's karyotype was 46,XY,der(9)t(8;9) (p21;p24)pat. A review shows that 6 additional patients with 46,XY sex reversal associated with monosomy of the distal short arm of chromosome 9 have been observed. The observation that all 7 patients with sex reversal share a deletion of the distal short arm of chromosome 9 is consistent with the hypothesis that the region 9p24 contains a gene or genes necessary for male sex determination. This present case narrows the chromosome interval containing a critical sex determination gene to the relatively small region 9p24. A molecular analysis of this region will provide a means to identify a gene involved in male sex determination. PMID- 9415693 TI - Trisomy 16pter to 16q12.1 and monosomy 22pter to 22q11.2 resulting from adjacent 2 segregation of a maternal complex chromosome rearrangement. AB - We report on a 16-week-old male fetus with partial trisomy 16 and partial monosomy 22 resulting from 3:3 adjacent-2 segregation of a maternal balanced complex chromosome translocation involving chromosomes 5, 16, and 22. The karyotype of the 29-year-old phenotypically normal mother was 46,XX,t(5;16;22)(q31.3;q12.1;q11.2). The karyotype of the fetus was 46,XY,der (5)t(5;16;22)(q31.3;q12.1;q11.2),+der(16) t(5;16;22)mat,-22. The fetus had multiple congenital anomalies, including bilateral cleft lip and palate. PMID- 9415694 TI - Complex congenital heart malformations in mosaic tetrasomy 8p: case report and review of the literature. AB - We describe a 5-month-old boy with complex congenital heart defects (dTGA, DORV, VSD, ASD, and PDA), minor facial and ear anomalies, deep palmar creases, multiple vertebral anomalies, agenesis of the corpus callosum, and mosaic tetrasomy 8p (47,XY,+i(8)(p10)[88%]/46,XY[12%] in blood with normal chromosomes in cultured skin fibroblasts. This infant represents the eleventh reported case of mosaic tetrasomy 8p since its first description by Kristofferson et al. [1988: Clin Genet 34:201-203]. The pattern of heart malformations and discordance of blood and fibroblast karyotypes make our case unique. Our report and review suggest that an important distinction between mosaic tetrasomy 8p and other chromosome 8 aneuploidies involves the increased incidence and complexity of congenital heart malformations. PMID- 9415695 TI - Prevalence of the factor V-Leiden mutation in four distinct American ethnic populations. AB - Resistance to activated protein C (APC) is the most common risk factor for venous thromboembolism, a major cause of morbidity and mortality with an incidence of about 1/1,000 per year. The Arg 506 to Gln mutation in exon 10 of the coagulation factor V gene (factor V-Leiden) has been found to be responsible for over 90% of the APC resistance cases and is an autosomal dominant trait. Initial studies have suggested that this mutation is restricted to individuals of European Caucasian extraction with an average allele frequency in European and American Caucasians of 4.4%, making it one of the most common monogenic disorders in the Caucasian population. A limited number of other ethnic populations have been tested and the mutation has been found only rarely. In our multiethnic survey of 602 individuals, Hispanic-Americans had the highest observed frequency of the factor V-Leiden mutant allele, 1.65%, while African-Americans had a somewhat lower frequency, 0.87%. No factor V-Leiden mutations were found in 191 Asian-Americans or 54 Native-Americans tested. These results indicate that the factor V-Leiden mutation segregates in populations with significant Caucasian admixture and is rare in genetically distant non-European groups. This ethnic stratification may be important in developing cost-effective selective screening programs to identify individuals at risk for thromboembolism and offer prophylactic therapy. PMID- 9415696 TI - Application of transmission disequilibrium tests to nonsyndromic oral clefts: including candidate genes and environmental exposures in the models. AB - Extensive epidemiological and genetic studies of the cause of oral clefts have demonstrated strong familial aggregation but have failed to yield definitive evidence of any single genetic mechanism. We used the transmission/disequilibrium test (TDT) to investigate the relationship between oral clefts and markers associated with five candidate genes by utilizing 160 parent-offspring trios. Conditional logistic regression models extended the TDT to include covariates as effect modifiers, thus permitting tests for gene-environment interactions. For four of these candidates [transforming growth factor alpha (TGFA), transforming growth factor beta 3 (TGFB3), retinoic acid receptor (RARA), and the proto oncogene BCL3], we detected modestly elevated odds ratios for the transmission of one marker allele to cleft probands when all the trios were analyzed together. These odds ratios increased when information on type of cleft, race, family history, or maternal smoking were incorporated as effect modifiers. We detected significant interaction between maternal smoking and the transmission of alleles for markers near TGFA and TGFB3; excess transmission of allele 3 at BCL3 was most significant among cleft lip probands; and the odds ratios for transmission of alleles at D19S178 and THRA1 were significant when ethnic group was included in the model. We suggest that utilizing an analytical strategy that allows for stratification of data and incorporating environmental effects into a single analysis may be more effective for detecting genes of small effect. PMID- 9415697 TI - First-trimester diagnosis of Blomstrand lethal osteochondrodysplasia. AB - Blomstrand chondrodysplasia is a rare lethal skeletal dysplasia with presumed autosomal-recessive inheritance. A family with 2 affected fetuses was studied. One fetus demonstrated a severe skeletal dysplasia at routine transabdominal ultrasound examination at 18.5 weeks of gestation. The pregnancy was terminated and the diagnosis of Blomstrand chondrodysplasia was made at autopsy. A second affected fetus was identified by first-trimester transvaginal ultrasound at 12 weeks of gestation. In this case the diagnosis was confirmed by posttermination radiography and histopathology. From these observations, Blomstrand chondrodysplasia seems like a lethal rhizo/mesomelic short-limb, early-onset dysplasia with autosomal-recessive inheritance. Easy detectability by transvaginal ultrasound is demonstrated, but general applicability awaits further studies on the intra- and interfamilial variability of this disorder. PMID- 9415698 TI - Trisomy 5q12-->q13.3 in a patient with add(13q): characterization of an interchromosomal insertion by forward and reverse chromosome painting. AB - We report on a patient with azoospermia, mild mental retardation, and minor physical anomalies. Chromosome analysis demonstrated the presence of additional material on the long arm of one chromosome 13. Forward chromosome painting using chromosome-specific libraries showed an insertion of material from chromosome 5. Further characterization with flow sorting of the aberrant chromosome and amplification by DOP-PCR followed by reverse chromosome painting showed specific trisomy of 5q12-->q13.3. PMID- 9415700 TI - Macrostomia, hypertelorism, atrophic skin, severe hypertrichosis without ectropion: milder form of Barber-Say Syndrome. PMID- 9415699 TI - Reactions to predictive testing in Huntington disease: case reports of coping with a new genetic status. AB - A predictive testing program for Huntington disease has been available in Stockholm, Sweden since October 1990. Psychosocial assessments were performed throughout the testing program to evaluate the impact of the risk situation itself and the effect of predictive testing, and to identify those individuals who were most vulnerable to severe stress and anxiety reactions. All subjects underwent neurological, neuropsychological, and psychiatric examinations. Individuals undergoing predictive testing were assessed twice by a genetic counsellor before receiving their results, and at 10 days (gene carriers only) and then 2, 6, 12, and 24 months after receiving the results. The process of coping with the test results and the psychological adjustment to knowledge about new genetic status have been shown to vary considerably. In this report, we describe the results obtained from two gene carriers and two noncarriers. The four persons chosen represent different ways of coping with the outcome of the test and of integrating knowledge about their genetic status into everyday life. These cases illustrate common themes and recurrent problems often surfacing during the counselling and testing process. The longitudinal evaluations provide information about the impact, adaptation, and long-term effects of living with a new genetic status. PMID- 9415701 TI - Leukocyte activation induces aryl hydrocarbon receptor up-regulation, DNA binding, and increased Cyp1a1 expression in the absence of exogenous ligand. AB - The aryl hydrocarbon receptor (AhR) functions as a transcription factor after ligand binding by halogenated aromatic hydrocarbons. 2,3,7,8-tetrachlorodibenzo-p dioxin (TCDD), the most toxic halogenated aromatic hydrocarbon, is dependent on binding to the AhR to mediate a broad range of toxic effects. Immune suppression is one of the most sensitive sequela associated with TCDD exposure, yet, paradoxically, resting leukocytes express a relatively low amount of AhR. Here we report that activation of leukocytes produced a 6-fold increase in AhR steady state mRNA levels and a concordant increase in AhR protein expression. Furthermore, leukocyte activation induced AhR translocation, DNA binding to a dioxin response element, and CYP1A1 transcription in the absence of TCDD. Activated leukocytes exhibited an even greater enhancement of dioxin response element binding by the AhR in the presence of TCDD than in the absence of TCDD. These studies suggest that the mechanism responsible for the sensitivity of immunocompetent cells to TCDD may be directly associated with a marked increase in AhR expression, which accompanies leukocyte activation. PMID- 9415702 TI - Molecular cloning and expression of an avian G protein-coupled P2Y receptor. AB - A family of G protein-coupled P2Y receptors that are activated by adenine and uridine nucleotides has been identified recently. Degenerate primers based on conserved sequences in these P2Y receptors were used to amplify turkey DNA, which was used to isolate the complete coding sequence of a cDNA that encodes a novel G protein-coupled receptor. Stable expression of this avian cDNA in 1321N1 human astrocytoma cells resulted in the conveyance of marked inositol phosphate responses to various nucleotides. Although this cloned avian receptor exhibited its highest homology to the previously cloned mammalian P2Y4 receptor, its pharmacological selectivity was not consistent with the avian receptor's being a species homologue of the P2Y4 receptor. That is, whereas the P2Y4 receptor is selectively activated by UTP and is not activated by ATP or Ap4A, the novel avian receptor was potently activated by ATP and Ap4A as well as by UTP. Taken together, these results describe the identification of an avian phospholipase C coupled P2Y receptor that, like the mammalian P2Y2 receptor, is activated by both adenine and uridine nucleotides. PMID- 9415703 TI - Coordinate regulation of stress- and mitogen-activated protein kinases in the apoptotic actions of ceramide and sphingosine. AB - We characterized participation of the stress-activated protein kinase (SAPK) cascade in the lethal actions of the cytotoxic lipid messengers ceramide and sphingosine in U937 human monoblastic leukemia cells. Acute exposure of U937 cells to either lipid resulted in loss of proliferative capacity, degradation of genomic DNA, and manifestation of apoptotic cytoarchitecture. Ceramide robustly stimulated p46-JNK1/p54-JNK2 activity and increased expression of c-jun mRNA and c-Jun protein; in contrast, sphingosine moderately stimulated p46-JNK1/p54-JNK2 and failed to modify c-jun/c-Jun expression. Dominant-negative blockade of normal c-Jun activity by transfection with the TAM-67 c-Jun NH2-terminal deletion mutant abolished the lethal actions of ceramide but was without effect on those of sphingosine, indicating that ceramide-related apoptosis is directly dependent on activation of c-Jun, whereas sphingosine-induced cell death proceeds via an unrelated downstream mechanism. Characterization of the mitogen-activated protein kinase (MAPK) cascade in these responses revealed a further functional disparity between the two lipids: basal p42-ERK1/ p44-ERK2 activity was gradually reduced by ceramide but immediately and completely suppressed by sphingosine. Moreover, blockade of the MAPK cascade by the aminomethoxyflavone MEK1 inhibitor PD-98059 unexpectedly activated p46-JNK1/p54-JNK2 and induced apoptosis in a manner qualitatively resembling that of sphingosine. Both lipids sharply increased p38 RK activity; selective pharmacological inhibition of p38-RK by the pyridinyl imidazole SB-203580 failed to mitigate the cytotoxicity associated with either ceramide or sphingosine, suggesting that p38-RK is not essential for lipid induced apoptosis. These findings demonstrate that reciprocal alterations in the SAPK and MAPK cascades are associated with the apoptotic influence of either lipid inasmuch as (i) ceramide-mediated lethality is primarily associated with strong stimulation of SAPK and weak inhibition of MAPK, whereas (ii) sphingosine mediated lethality is primarily associated with weak stimulation of SAPK and strong inhibition of MAPK. We therefore propose that leukemic cell survival depends on the maintenance of an imbalance of the outputs from the MAPK and SAPK systems such that the dominant basal influence of the MAPK cascade allows sustained proliferation, whereas acute redirection of this balance toward the SAPK cascade initiates apoptotic cell death. PMID- 9415704 TI - Characterization of a novel bisacridone and comparison with PSC 833 as a potent and poorly reversible modulator of P-glycoprotein. AB - Novel compounds, composed of two acridone moieties connected by a propyl or butyl spacer, were synthesized and tested as potential modulators of P-glycoprotein (P gp)-mediated multidrug resistance. The propyl derivative 1,3-bis(9-oxoacridin-10 yl)-propane (PBA) was extremely potent and, at a concentration of 1 microM, increased steady state accumulation of vinblastine (VLB) approximately 9-fold in the multidrug-resistant cell line KB8-5. In contrast to the readily reversible effects of VRP and cyclosporin A on VLB uptake and similar to the effects of the cyclosporin analog PSC 833, this modulation by PBA was not fully reversed 6-8 hr after transfer of cells to PBA-free medium. Continuous exposure to 3 microM PBA was nontoxic and could completely reverse VLB resistance in KB8-5 cells. Consistent with its effects on VLB transport, the drug resistance-modulating effect of PSC 833 was significantly more persistent than that of VRP. However, the effect of PBA was, like that of VRP, rapidly reversed once the modulator was removed from the extracellular environment. PBA was able to compete with radiolabeled azidopine for binding to P-gp and to stimulate P-gp ATPase activity. However, both the steady state accumulation of PBA and the rate of efflux of PBA were similar in drug-sensitive KB3-1 and drug-resistant KB8-5 cells, suggesting that this compound is not efficiently transported by P-gp. These results indicate that PBA represents a new class of potent and poorly reversible synthetic modulators of P-gp-mediated VLB transport. PMID- 9415705 TI - Characterization of a human glutathione S-transferase mu cluster containing a duplicated GSTM1 gene that causes ultrarapid enzyme activity. AB - The mu class glutathione S-transferase gene GSTM1 is polymorphic in humans, with approximately half of the Caucasian population being homozygous deleted for this gene. GSTM1 enzyme deficiency has been suggested to predispose people to lung and bladder cancer. Some people in a Saudi Arabian population, however, have been described previously with ultrarapid GSTM1 enzyme activity. Here we have evaluated the molecular genetic basis for this observation. Genomic DNA from two Saudi Arabian subjects exhibiting ultrarapid enzyme activity and from 13 Swedish subjects having null, one, or two GSTM1 genes were subjected to restriction fragment length polymorphism analysis using the restriction enzymes EcoRI, EcoRV, and HindIII and combinations thereof. Hybridization was carried out using a full length GSTM1 cDNA or the 5' and 3' parts of the cDNA. The restriction mapping data revealed the presence of a GST mu cluster with two GSTM1 genes in tandem situated between the GSTM2 and GSTM5 genes. A quantitative multiplex polymerase chain reaction method, which simultaneously amplified a fragment of the GSTM1 gene and the beta-globin gene, was developed, and the genomic GSTM1 copy number was determined from the GSTM1/beta-globin ratio. This method clearly separated GSTM1 +/- subjects (ratios between 0.4 and 0.7) from GSTM1 +/+ subjects (ratios between 0.8 and 1.2). The two Saudi Arabians with ultrarapid GSTM1 activities had ratios of approximately 1.5, indicating that they carried three GSTM1 genes. These results demonstrate the existence of a novel mu class GST cluster containing a duplicated active GSTM1 gene causing ultrarapid enzyme activity. PMID- 9415706 TI - Regulation of m2 muscarinic receptor gene expression by platelet-derived growth factor: involvement of extracellular signal-regulated protein kinases in the down regulation process. AB - To study the role of mitogen-activated protein kinase in the regulation of M2 receptors, we studied the effect of platelet-derived growth factor (PDGF) on M2 receptor gene expression. PDGF (4 ng/ml) caused a time-dependent decrease in M2 receptor number and in m2 receptor mRNA levels in HEL 299 cells. The PDGF-induced loss in m2 mRNA required de novo protein synthesis and occurred through a decrease in the rate of transcription of the m2 receptor gene. The down regulation of M2 receptors was not accompanied by an uncoupling of the remaining receptors, indicating a large receptor reserve in these cells. Preincubations with the phosphatidylinositol 3-kinase inhibitor wortmannin, the protein kinase C inhibitor GF 109203X and the cAMP-dependent protein kinase inhibitor H-8 did not attenuate PDGF-induced down-regulation, indicating a lack of involvement of these enzymes in the down-regulation process. Activation of the extracellular signal regulated protein kinase (ERK) 1 and 2 proteins was measured by an "in gel" phosphorylation assay. Carbachol did not activate ERK1 or 2, whereas PDGF and 4 beta-phorbol 13,14-dibutyrate resulted in a large increase in ERK1 and 2 activity along with a decrease in m2 mRNA. Preincubation with PD 098059, an inhibitor of mitogen-activated protein kinase kinase, inhibited PDGF- and 4 beta-phorbol 13,14 dibutyrate-mediated activation of ERK 1 and 2 in a concentration-dependent manner. The inhibitory action of PD 098059 was reflected at the mRNA level attenuating both PDGF- and 4 beta-phorbol 13,14-dibutyrate-mediated decreases in m2 mRNA. These results suggest a role of ERK1 and 2 in the regulation of muscarinic m2 receptor gene expression. PMID- 9415707 TI - Identification of benzo[a]pyrene-inducible cis-acting elements within c-Ha-ras transcriptional regulatory sequences. AB - Previous studies in this laboratory have demonstrated that transcriptional deregulation of c-Ha-ras expression is associated with the induction and maintenance of proliferative vascular smooth muscle cell (SMC) phenotypes by benzo[a]pyrene (BaP). We examined previously undescribed cis-acting elements within the proximal 5' regulatory region of c-Ha-ras (-550 to +220) for their ability to influence BaP-induced transcription in murine SMCs. BaP-inducible DNA binding activity was demonstrated at a site located -30 relative to the major start site cluster at +1 that exhibits extensive homology to a consensus aryl hydrocarbon response element (AHRE), as well as a site located at -543 that contains a consensus electrophile response element (EpRE). In vitro cross-linking studies revealed the specific interaction of 104- and 96-kDa proteins with the putative AHRE and of an 80-kDa protein with the EpRE. The use of monoclonal antibodies to the aryl hydrocarbon receptor transcription factor in competition electrophoretic mobility shift assays indicated this protein is specifically induced by BaP to interact at the AHRE within the c-Ha-ras 5' regulatory region. Transient transfection with an Ha-ras promoter construct containing the putative AHRE but lacking the EpRE linked to the chloramphenicol acetyl transferase reporter gene, followed by challenge with BaP (0.3, 3.0, and 30 microM), revealed transcriptional activation that was not statistically significant. However, insertion of an oligonucleotide composed of the EpRE immediately upstream of basal sequences at -330 was associated with strong activation of transcription by BaP. These data indicate that c-Ha-ras gene expression is modulated by BaP via a complex mechanism that likely involves interactions among multiple regulatory elements. We conclude that c-Ha-ras expression is regulated by BaP at the transcriptional level, a response that may constitute an epigenetic basis of atherogenesis. PMID- 9415708 TI - Naloxone activation of mu-opioid receptors mutated at a histidine residue lining the opioid binding cavity. AB - The mu-opioid receptor is the principal site of action in the brain by which morphine, other opiate drugs of abuse, and endogenous opioid peptides effect analgesia and alter mood. A member of the seven-transmembrane domain (TM) G protein-coupled receptor (GPCR) superfamily, the mu-opioid receptor modulates ion channels and second messenger effectors in an opioid agonist-dependent fashion that is reversible by the classic opiate antagonist naloxone. Mutation of a histidine residue (His297) in TM 6 afforded agonist-like G protein-coupled signal transduction mediated by naloxone and other alkaloid antagonists and enhanced the intrinsic activity of documented alkaloid partial agonists, including buprenorphine. The intrinsic activities of all opioid peptide agonists and antagonists tested were not altered at the His297 mutant receptors. Consistent with a role for the TM 6 histidine in maintaining high affinity binding sites for opioid agonists and antagonists, opioid ligand-dependent protection of this residue from a histidine-specific alkylating agent indicated that the His297 side chain is positioned in or very near the binding cavity. The TM 6 His297 mutants identify a discrete region of the receptor critical for determining whether a specific drug pharmacophore triggers receptor activation. Because many GPCRs possess a similarly positioned TM histidine residue, our findings with the mu opioid receptor may extend to these receptors and potentially serve as a model for rational design of therapeutic GPCR partial agonists and antagonists. PMID- 9415709 TI - Enhanced stimulatory adenylyl cyclase signaling during opioid dependence is associated with a reduction in palmitoylated Gs alpha. AB - Chronic opioid treatment of stably mu-opioid receptor transfected human mammary epidermoid A431 carcinoma cells (clone A431/mu 13) results in sensitization of adenylyl cyclase (AC), a cellular adaptation associated with drug dependence. Up regulation of AC is characterized by significantly increased levels of both basal and post-receptor-stimulated effector activities, which develop without any apparent change in the quantity of stimulatory G proteins and the maximum catalytic activity of AC. Here, we report that detergent extracts from membranes of chronically morphine-treated (10 microM; 2 days) A431/mu 13 cells display higher stimulatory AC activities as assessed in the S49cyc- reconstitution assay. This finding is most likely due to an increased functional activity of Gs alpha because the addition of exogenous G beta gamma subunits, which per se stimulate AC in S49cyc- membranes, failed to affect the difference in reconstitutive AC activity. Moreover, both chemical depalmitoylation by hydroxylamine and inhibition of palmitoyl-CoA transferase in vivo by tunicamycin treatment incresed the reconstitutive activity of detergent extracts and eliminated the differences between native and opioid-dependent cells, indicating that the increase in stimulatory activity is due to depalmitoylation of Gs alpha. Indeed, metabolic labelling studies with [3H]palmitic acid revealed that chronic opioid treatment reduces considerably the fraction of palmitoylated Gs alpha in the plasma membrane. Furthermore, high affinity [3H]forskolin binding experiments demonstrated that depalmitoylated Gs alpha is able to associated directly with AC during the state of opioid dependence even without preceding receptor activation. These results suggest that post-translational palmitoylation of Gs alpha provides a potential regulator of transmembrane signaling. Moreover, accumulation of the depalmitoylated form of Gs alpha in the plasma membrane as reported herein may contribute to the increase in stimulatory AC signaling, as is characteristic for the state of opioid dependence. PMID- 9415711 TI - Both the cyclic AMP response element and the activator protein 2 binding site mediate basal and cyclic AMP-induced transcription from the dominant promoter of the rat alpha 1B-adrenergic receptor gene in DDT1MF-2 cells. AB - cAMP markedly increases alpha 1B adrenergic receptor (alpha 1B-AR) expression in FRTL-5 and PC C13 rat thyroid cells, DDT1MF-2 smooth muscle cells, primary rat hepatocytes, and K9 rat liver cells. Here, we used DDT1MF-2 cells to evaluate further the mechanisms by which cAMP stimulates alpha 1B-AR expression. Receptor binding assays, Northern blotting, and nuclear run-on analyses demonstrated that forskolin (1 microM) in the presence of isobutylmethylxanthine (0.25 mM) increased alpha 1B-AR numbers, mRNA level, and gene transcription rate by 2.3 +/- 0.2-, 2.5 +/- 0.3-, and 3.5 +/- 0.2-fold over control, respectively. Dibutyryl cAMP (1 mM) plus isobutylmethylxanthine (0.25 mM) also enhanced alpha 1B-AR density by 2.7 +/- 0.1-fold over control. Further experiments demonstrated that the induction of alpha 1B-AR by forskolin requires new protein synthesis and is protein kinase A dependent. In DDT1MF-2 cells transfected with alpha 1B-AR gene P2 promoter/CAT constructs, both forskolin and dibutyryl cAMP significantly increased P2 promoter activity. The P2 promoter region of the rat alpha 1B-AR gene (-813 to -432) contains a cAMP response element (CRE) (-444 to -437) and an AP2 binding site (-647 to -638). Mutations in either one of these elements alone led to a decrease in both basal and cAMP-induced P2 promoter activity. Mutations in both elements caused a further inhibition of basal transcription and a complete block of cAMP-induced P2 promoter activity. Direct binding of purified activator protein 2 (AP2) to the AP2 element in the P2 promoter was reported previously. Gel mobility shift and super-shift assays using liver nuclear extracts from either rat liver or DDT1MF-2 cells demonstrated that the CRE in the alpha 1B-AR gene bound CRE binding protein. These data indicate that both the CRE and the AP2 element in the P2 promoter contribute to basal as well as cAMP induced transcription of the alpha 1B-AR gene in DDT1MF-2 cells. PMID- 9415710 TI - Proximal nephron Na+/H+ exchange is regulated by alpha 1A- and alpha 1B adrenergic receptor subtypes. AB - Activation of alpha 1-adrenergic receptors (alpha 1-AR) increases Na+/H+ exchange (NHE) in proximal tubule. NHE mediates the majority of active Na+ absorption in the proximal tubule. Three alpha 1-AR subtypes have been detected in kidney by molecular and binding techniques. We detected message for all three alpha 1-AR subtypes in mouse proximal tubule cells through reverse transcription-polymerase chain reaction and Northern analysis. To determine the alpha 1-AR subtypes that regulate NHE in mouse proximal tubule cells, two strategies were used: (i) antisense oligodeoxynucleotides (ODNs) to selectively inhibit expression of alpha 1A-, alpha 1B-, and alpha 1D-AR subtypes and (ii) subtype-selective alpha 1-AR antagonists. Streptolysin-O permeabilization was used to introduce antisense and sense ODNs into cells three times over 72 hr. Western blot analysis of membranes prepared from cells treated with alpha 1B-AR antisense ODN demonstrated that alpha 1B-AR protein expression was reduced by 90% at 72 hr compared with control or sense ODN treatments. Functional regulation of NHE by alpha 1-ARs was determined by alpha 1-AR agonist changes in intracellular pH (pHi) in cells grown on coverslips and loaded with 2',7'-bis(2-carboxyethyl)-5(6)carboxyfluorescein acetoxymethyl ester. Antisense ODNs for alpha 1B-AR significantly reduced phenylephrine (PHE)-induced maximal changes in pHi by 49%. The PHE-induced changes in pHi observed in cells treated with alpha 1A-AR antisense ODNs was reduced by 42%. The selective alpha 1A-AR antagonist WB-4101 and the alpha 1B-AR antagonist spiperone reduce PHE-induced pHi increases to a comparable extent. No significant changes in pHi were observed with cells treated with alpha 1D-AR antisense ODNs or the alpha 1D-AR antagonist BMY 7378 compared with untreated cells. Combined treatment with alpha 1A- and alpha 1B-AR antisense ODNs and antagonists additively inhibits PHE-induced delta pHi by 90%. We conclude that alpha 1A and alpha 1B-AR but not alpha 1D-ARs regulate NHE in proximal tubule cells. PMID- 9415712 TI - Neuropeptide Y inhibits chromaffin cell nicotinic receptor-stimulated tyrosine hydroxylase activity through a receptor-linked G protein-mediated process. AB - Acetylcholine stimulation of bovine chromaffin cells results in increased norepinephrine and epinephrine secretion accompanied by a corresponding increase in synthesis. The addition of neuropeptide Y (NPY) to the culture medium prevents the increase in catecholamine synthesis but not secretion. Treatment of chromaffin cells with nicotine produces a concentration-dependent increase in tyrosine hydroxylase activity (IC50 = 1.2 microM) that is reduced if NPY is present during stimulation. Tyrosine hydroxylase activity decreases in a concentration-dependent fashion if increasing amounts of NPY are included in the culture medium, IC50 = 0.2 nM. Treatment with pertussis toxin completely prevents the effect of NPY. The rank order of potency for inhibition of tyrosine hydroxylase activity is NPY > or = [Leu31,Pro34]NPY > or = peptide YY > NPY2-36 > NPY13-36 > NPY18-36 > or = NPY26-36 >> NPY1-30, suggesting a NPY-Y1 receptor subtype. Examination of the effect of NPY on nicotine stimulation of chromaffin cell protein phosphorylation showed that NPY produces a concentration-dependent decrease in a 60-kDa protein, IC50 = 6.4 nM. The effect of NPY is pertussis toxin sensitive. The rank order of potency is [Leu31,Pro34]NPY > or = NPY >> NPY18-36. Immunoprecipitation confirmed the identity of the 60-kDa protein as tyrosine hydroxylase. PMID- 9415713 TI - A combination of mutations in the CYP2D6*17 (CYP2D6Z) allele causes alterations in enzyme function. AB - In many black African populations, the capacity for CYP2D6-dependent drug metabolism is generally reduced. A specific variant of the CYP2D6 gene (CYP2D6*17) that carries three functional mutations (T107I, R296C, and S486T) has been found to be present in Zimbabwean subjects with impaired CYP2D6-dependent hydroxylase activity. To evaluate whether the CYP2D6*17 allele was the major cause behind the decreased rate of drug metabolism and to examine the role of the different mutations, CYP2D6 cDNAs containing all eight combinations of the mutations were created. Expression of the cDNAs in COS-1 cells revealed that the CYP2D6 17 enzyme displayed only 20% of the wild-type (CYP2D6 1) activity, whereas the T107I substitution on its own had no significant effect on enzyme function. Expression in yeast showed that the three possible single amino-acid mutant CYP2D6 variants all had properties similar to CYP2D6 1 when the kinetics of bufuralol hydroxylation was examined. However, enzymes containing both the T107I and R296C mutations exhibited a more than 5-fold higher K(m) for bufuralol than the wild-type enzyme, whereas the S486T mutation was of little importance. In contrast, when codeine was used as a substrate, the T107I substitution alone was sufficient to cause a significant increase in the apparent K(m), indicating a differential effect for this substitution depending on the CYP2D6 substrate. In conclusion, the CYP2D6*17 allele represents the first human cytochrome P450 polymorphic variant in which a combination of substitutions is required to alter the enzyme's catalytic properties and is the first case in which a decreased CYP2D6 activity, as monitored in vivo, has been documented to be caused by an enzyme with altered affinity for CYP2D6 substrates. PMID- 9415715 TI - Antisense inhibition of 5-hydroxytryptamine2a receptor induces an antidepressant like effect in mice. AB - Treatment with different antidepressants is invariably accompanied by the down regulation of the 5-hydroxytryptamine2A (5-HT2A) receptor. To determine whether receptor down-regulation is an essential part of antidepressant action, we manipulated levels of the 5-HT2A receptor by using a nonpharmacological approach. Here, we report that down-regulation of the 5-HT2A receptor by intracerebroventricular injection of antisense oligonucleotides resulted in an antidepressant-like effect in mice. Animals with 5-HT2A receptor deficiency showed less immobility in the Porsolt's forced swim test, a well established animal model that is used to identify drugs with an antidepressant effect. The overall locomotor activity of the receptor-deficient animals was not altered, demonstrating the specificity of the behavioral change in the Porsolt's forced swim test. Reduced immobility in this test was accompanied by a greater c-Fos response in piriform cortex. Because 5-HT2A receptors have been localized on gamma-aminobutyric acid interneurons, the inhibitory activity of these neurons may be impaired at low receptor levels, leading to a greater c-Fos response in the piriform cortex and increased mobility in the Porsolt's forced swim test. These experiments demonstrate that down-regulation of the 5-HT2A receptor alone is sufficient to achieve an antidepressant-like effect in mice and suggest that receptor down-regulation may be an essential part of the antidepressant drug action. PMID- 9415714 TI - Potent inhibitors of human immunodeficiency virus type 1 integrase: identification of a novel four-point pharmacophore and tetracyclines as novel inhibitors. AB - A four-point pharmacophore was constructed from energy-minimized structures of chicoric acid and dicaffeoylquinic acid. The search of 206,876 structures in the National Cancer Institute 3D database yielded 179 compounds that contain this pharmacophore. Thirty-nine of these compounds were tested in an in vitro assay specific for human immunodeficiency virus type 1 integrase (IN). Each retrieved structure was fit to the pharmacophore, and the conformation that afforded the best fit was identified. Twenty of the 39 compounds tested exhibited IC50 values of < 20 microM. Among the most potent inhibitors, tetracyclines emerged as a new class of inhibitors. Although the parent tetracycline exhibited marginal potency against purified IN, all substituted tetracyclines tested showed 5-100-fold increased potency. Disintegration assays with truncated IN mutants indicated that tetracyclines inhibit the IN catalytic core domain. To investigate whether chelation of divalent metals is implicated in differential potency of tetracyclines, enzyme assays were performed in the presence of both Mn2+ or Mg2+; no significance difference in potency was observed. Rolitetracycline inhibited IN/DNA complex formation in the presence of EDTA, which suggests that inhibition was metal independent. Rolitetracycline reversed DNA binding of IN after the complex was allowed to form before the addition of drug. Selectivity of tetracyclines was also examined in an assay specific for topoisomerase I, and none of the tetracyclines tested induced topoisomerase I-mediated cleavable complex or inhibited camptothecin-induced cleavable complex. Remarkable potency against the IN in the absence of divalent metals and the core enzyme coupled with water solubility makes tetracyclines potential candidates for X-ray crystal structure determination with IN. PMID- 9415716 TI - Role of G alpha q or G alpha o proteins in alpha 1-adrenoceptor subtype-mediated responses in Fischer 344 rat aorta. AB - Previous studies showed that alpha-adrenoceptor (AR) stimulation with norepinephrine is more potent at eliciting contraction in aortas from 1-month-old Fischer 344 rats than from older rats and that this response is mediated by alpha 1b- and alpha 1d-AR subtypes in 1-month-old rats. We examined the G proteins responsible for alpha 1-AR-mediated contractile response and inositol phosphate accumulation in the aortas of 1-month-old Fischer 344 rats. Pertussis toxin (PTX) treatment (2.5 micrograms/ml for 4 hr) of aortic rings partially inhibited phenylephrine (PHE)-stimulated contraction and inositol phosphate accumulation, suggesting the involvement of PTX-sensitive and -insensitive G proteins. Specific antisera directed against G alpha q and G alpha o but not G alpha s and G alpha i precipitated specific alpha 1-AR binding sites labeled with 2-[beta-(4-hydroxy-3 [125I]iodophenyl)ethylaminomethyl]tetralone. The number of 2-[beta-(4-hydroxy-3 [125I]iodophenyl)ethylaminomethyl]tetralone binding sites precipitated by G alpha proteins was increased by activating membrane alpha 1-ARs with PHE. Moreover, PHE stimulated the palmitoylation of G alpha q and G alpha o, and this response was blocked by the alpha 1-AR antagonist prazosin. Characterization of the alpha 1-AR subtypes that couple to G proteins indicates that although aortic alpha 1a-, alpha 1b-, and alpha 1d-ARs were associated with G alpha q, alpha 1b-AR was also linked to G alpha o. These results suggest that alpha 1-ARs mediate the contractile response in rat aorta by coupling to both Gq protein and the PTX sensitive G(o) protein. PMID- 9415717 TI - Distribution of mRNA encoding three alpha 2-adrenergic receptor subtypes in the developing mouse embryo suggests a role for the alpha 2A subtype in apoptosis. AB - alpha 2-Adrenergic receptors (alpha 2-ARs) respond to norepinephrine and epinephrine to mediate diverse physiological effects. Using in situ hybridization, the expression pattern of the mRNA encoding the three alpha 2-AR subtypes (alpha 2A, alpha 2B, and alpha 2C) was examined in the mouse embryo. The mRNA encoding the three subtypes was first detected at stage 9.5 days postcoitus (d.p.c.) for the alpha 2A-AR (coincident with norepinephrine availability), 11.5 d.p.c. for the alpha 2B-AR, and 14.5 d.p.c. for the alpha 2C-AR subtype. The mRNA encoding the alpha 2A-AR subtype shows both the earliest and the most widespread expression pattern, including developing stomach and cecum, many craniofacial regions and areas in the central nervous system. Strikingly, the alpha 2A-AR mRNA is expressed in the interdigital mesenchyme between stage 12.5 and 14.5 d.p.c. in parallel with digit separation, raising the possibility that the alpha 2A-AR might contribute to the apoptotic events underlying this process. To test whether alpha 2A-AR can signal apoptotic events, the alpha 2A-AR subtype was introduced into two mouse mesenchymal cell lines, C3H/10t1/2 and NIH-3T3; expression of the alpha 2A-AR correlated with accelerated apoptosis, as detected both by the TUNEL assay and the loss of cell viability. In contrast to the wide distribution of mRNA encoding the alpha 2A-AR subtype, the alpha 2B-AR mRNA was detected only in the developing liver and was most readily detectable between 11.5 and 14.5 d.p.c., when the liver is the principal site of hematopoiesis. The alpha 2C-AR mRNA is detected in the nasal cavity and cerebellar primordium only at > or = 14.5 d.p.c. These studies represent the first characterization of the temporal and spatial expressions of the alpha 2A-AR, alpha 2B-AR, and alpha 2C-AR subtypes during embryogenesis and provide important insights concerning the loci and possible roles of alpha 2-AR-mediated regulation of physiological processes during the developmental program. PMID- 9415718 TI - In vivo production of nitric oxide in rats after administration of hydroxyurea. AB - The metabolism of nitrovasodilators such as glyceryl trinitrate and nitroprusside provides the active moiety of these drugs (that is, nitric oxide). This process is not limited to the known nitrovasodilators, but also occurs with nitroaromatic antimicrobials. Here we report that the administration of hydroxyurea, an antitumor drug, to rats at pharmacological doses formed detectable nitrosyl hemoglobin, which increased with dose. At higher doses, nitrosyl hemoprotein complexes could also be detected in liver tissue. [15N]hydroxyurea was synthesized and compared with [14N]hydroxyurea. These observations verified that nitric oxide detected as nitrosyl hemoglobin or nitrosyl hemoprotein complexes in rats was the result of the metabolism of hydroxyurea. The time course and dose dependence of nitric oxide generation were also investigated. Hydroxyurea's antineoplastic activity is caused by its direct action on ribonucleotide reductase, the rate-limiting enzyme in DNA synthesis. Because nitric oxide also inhibits ribonucleotide reductase, this metabolite may supplement this action of hydroxyurea. In addition, the known ability of hydroxyurea to ease the pain of sickle cell anemia patients may be the result of vasodilation by the drug-derived nitric oxide. PMID- 9415719 TI - Site-directed mutagenesis on the m2 muscarinic acetylcholine receptor: the significance of Tyr403 in the binding of agonists and functional coupling. AB - The first step in the transmembrane signal mediated by G protein-coupled receptors is binding of agonist to receptors at the cell surface. The mechanism of the resulting receptor activation is not clear, but models based on the ternary complex model are capable of explaining most of the observations that have been reported in G protein-coupled receptors. This model suggests that a single agonist/receptor/G protein complex capable of activating G protein is formed as the result of agonist binding. Extensions of this basic model differ primarily in whether an equilibrium between active and inactive conformations is required to explain experimental results. We report results on ligand binding and coupling to physiological effector systems of the m2 muscarinic acetylcholine receptor site-directed mutant Y403F (residue 403 mutated from tyrosine to phenylalanine) expressed in Chinese hamster ovary cells and compare our results with results reported for the homologous Y506F mutation in the m3 muscarinic receptor [J. Biol. Chem. 267:19313-19319 (1992)]. The mutation in the m2 muscarinic receptor reduced absolute agonist affinities more dramatically than in the m3 muscarinic receptor. Unlike the results reported for the m3 subtype mutant, in which coupling to physiological effector systems was reduced, coupling to effector systems for the mutant in the m2 subtype was robust. In the Y403F m2 muscarinic receptor, the difference between the two agonist binding affinities was greater than in the wild-type receptor, whereas in the m3 subtype, the effect of the mutation was to decrease this difference. A prediction of the ternary complex model is that relative binding affinities will affect the steady state concentration of the agonist/receptor/G protein complex and, as the result, the extent of G protein coupling. These results can best be rationalized by this model, which suggests that the activation of G protein-coupled receptors is achieved by the relative affinity of agonist for two receptor states and does not require the existence of multiple states in conformational equilibrium. PMID- 9415720 TI - Voltage-dependent inhibition of N- and P-type calcium channels by the peptide toxin omega-grammotoxin-SIA. AB - We studied the mechanism by which the peptide omega-grammotoxin-SIA inhibits voltage-dependent calcium channels. Grammotoxin at concentrations of > 50 nM completely inhibited inward current carried by 2 mM barium through P-type channels in rat cerebellar Purkinje neurons when current was elicited by depolarizations up to +40 mV. However, outward current (carried by internal cesium) elicited by depolarizations to > +100 mV was either unaffected or enhanced in the presence of toxin. Tail current activation curves showed that grammotoxin shifted the steady state voltage dependence of channel activation by approximately +40 mV. Activation in the presence of toxin was far slower in addition to having altered voltage dependence. Grammotoxin also inhibited N-type calcium channels in rat and frog sympathetic neurons, with changes in channel voltage dependence and kinetics nearly identical to those of P-type channels. Experiments with monovalent ions as the only charge carriers showed that toxin effects on channel activation and kinetics depended on voltage, not on direction of current flow or on the current-carrying ion. Repeated trains of large depolarizations relieved toxin inhibition, as if toxin affinity for activated channels were low. The effects of grammotoxin on gating of P-type channels are very similar to those of omega-Aga-IVA, but combined application of the two toxins showed that grammotoxin binding is not prevented by saturating binding of omega-Aga-IVA. We conclude that grammotoxin potently inhibits both P-type and N type channels by impeding channel gating and that grammotoxin binds to distinct or additional sites on P-type channels compared with omega-Aga-IVA. PMID- 9415721 TI - Functional deactivation of the major neuronal nicotinic receptor caused by nicotine and a protein kinase C-dependent mechanism. AB - The effect of nicotine on the major human neuronal nicotinic receptor (alpha 4 beta 2 subtype) was studied in permanently transfected HEK 293 cells. Prolonged exposure to low concentrations of nicotine (1 microM) increased epibatidine binding but functionally deactivated the nicotinic receptor, abolishing Ca2+ influx in response to an acute nicotine challenge. Deactivation could also be caused by down-regulating protein kinase C (PKC) activity with 0.5 microM phorbol 12,13-dibutyrate or briefly incubating cells with the PKC inhibitor NPC-15437. Recovery from receptor deactivation caused by either nicotine treatment or PKC inhibition occurred slowly (4-6 hr). Reversal of nicotine-induced deactivation was accelerated by the addition of inhibitors of protein phosphatases 2A and 2B. These data suggest a hypothetical mechanism of nicotine-induced deactivation that involves dephosphorylation of nicotinic receptors at PKC phosphorylation sites. PMID- 9415722 TI - Finger joint reconstruction after mutilation of the hand. PMID- 9415723 TI - The hung up shoulder: anterior subluxation locking in abduction. AB - The hung up shoulder, or anterior subluxation locking in abduction, is a bizarre clinical picture which is not frequently seen. It is probably due to the fact that, in the subluxed position, the subscapularis muscle becomes an abductor, rather than an internal rotator. External rotation might shift the subscapular muscle fibres towards the proximal aspect of the humeral head, while joint laxity favors subluxation. It is possible that the hung up shoulder is just one aspect of multidirectional shoulder instability, given the tendency to generalised joint laxity, the frequent autoreduction, the positive sulcus sign, and initiation of subluxation by either abduction-external rotation or extension. In this series three out of four patients were treated conservatively and performed well in daily life; however, only the fourth patient had almost unlimited access to sports, thanks to surgical stabilization. PMID- 9415724 TI - Operative treatment of humeral shaft fractures. AB - The results of the operative treatment of 27 humeral shaft fractures treated at the University of Louisville during a 2-year period were reviewed. The aim of this study was to analyze 1) the indications and results of surgical treatment, 2) the indications for nailing versus plating, and 3) the failures and their treatment (especially surgical nonunions). Indications for surgery were polytrauma patients (including open fractures, associated neurovascular injuries, associated ipsilateral forearm injuries) and isolated unstable fractures in which closed reduction failed. Plate and screw osteosynthesis was used in patients with proximal and distal fractures, in the presence of neurovascular injuries, progressive radial nerve palsy and failure of closed reduction due to interposition of soft tissue. Intramedullary antegrade nailing was preferentially used in polytrauma patients. Seven patients (25%) needed further surgery because of nonunion. The frequency was higher after plating (30%) than after nailing (20%), it was more common in comminuted fractures, middle third fractures and after insufficient distal locking. Exchange nailing resulted in union in 5 of the 7 cases. Although excellent results with low complication rates are reported in the recent literature following plate and screw osteosynthesis or locked intramedullary nailing, we found that operative treatment of difficult humeral shaft fractures is still fraught with a high complication rate. PMID- 9415725 TI - Extensor indicis proprius to extensor pollicis longus transfer: results and complications. AB - In a detailed analysis of hand, thumb and index function after extensor indicis proprius transfer for extensor pollicis longus rupture in 13 patients, we found 12 excellent or good results despite mobility restriction in most of them. The extension lag in the metacarpophalangeal and interphalangeal joint of the thumb and the metacarpophalangeal joint of the index was respectively 17 degrees, 8.5 degrees, and 25 degrees. Extension force of the index was reduced to 65%. Grip force and dexterity reached 90% and 115% of the contralateral side. PMID- 9415726 TI - Application of the IFSSH(3)-classification for congenital anomalies of the hand; results and problems. AB - The extended classification proposed by the IFSSH was used to classify 1013 hand anomalies in 925 hands of 650 patients. We found associated anomalies in 26.7%. The classification was straightforward in 86%, difficult in 6.6% and not possible in 7.8%. In group I the radial and ulnar deficiencies, limited to the hand and without forearm deficiencies should be included. Group II was the most important group including 513 anomalies. We propose to include in this group the Madelung deformity, the Kirner deformity and congenital trigger fingers and trigger thumbs. Triphalangeal thumbs are a problem, we suggest to list this anomaly in group III and to consider it as a duplication in length. It is not always possible to evaluate the (transverse) absence of the fingers or hand. Longitudinal deficiencies (group IIB), symbrachydactyly group (V) and amniotic bands (group VI) occasionally develop a phenotype similar to the genuine transverse deficiency (group IA). PMID- 9415727 TI - Total knee arthroplasty in the young rheumatoid patient. AB - Thirty-four total knee arthroplasties were performed for severe rheumatoid arthritis in 25 patients younger than 45 years. All patients were available for follow-up evaluation at an average of 7.2 years. According to the Knee Society scoring system, the knee score improved from an average of 21 points preoperatively to 85 points at follow-up (p < 0.001). The average functional score improved from 23 points to 87 points (p < 0.001). Average range of motion improved from 71 degrees to 93 degrees (p < 0.001). Nonprogressive radiolucencies less than 1-mm thick were observed in 6 knees. One knee was revised for severe polyethylene wear; another case was revised for chronic patellar dislocation. Actuarial survivorship analysis estimates a 97% survivorship after 5 years and 90% after after 10 years. In young rheumatoid patients, total knee arthroplasty can therefore be considered as a reliable procedure, with satisfactory results during at least the first 5 to 10 postoperative years. PMID- 9415728 TI - Juvenile kyphosis: effects of different variables on conservative treatment outcome. AB - We reviewed the records of 212 patients with idiopathic or Scheuermann-type juvenile kyphosis (Scheuermann's disease). The 200 patients available for follow up were divided into three groups depending on the degree of angular deformity, and the influence of different variables on treatment outcome in each group was investigated. A very influential positive variable was combined treatment with a body cast plus brace; exercise treatment also produced acceptable results. Other variables that positively influenced the outcome of treatment were compliance with treatment, and (unexpectedly) elevated initial Risser sign (skeletal maturity). An initial Risser sign of 0 or 1 was, in contrast with other studies, associated with smaller improvement. However, initial maximal wedging, etiology and initial assessment of curve flexibility did not influence the degree of improvement in the initial angular deformity. PMID- 9415729 TI - Tri-calcium phosphate ceramics and allografts as bone substitutes for spinal fusion in idiopathic scoliosis as bone substitutes for spinal fusion in idiopathic scoliosis: comparative clinical results at four years. AB - The authors present the results of a comparative study of two series of posterolateral arthrodeses for scoliosis performed using COTREL DUBOUSSET instrumentation. Fifty-four consecutive patients underwent surgery for idiopathic scoliosis using the same technique. Thirty received a graft consisting of a mixture of corticocancellous autologous and allogenic bone frozen at -80 degrees, and 24 patients were grafted with a mixture of cortico-cancellous autologous bone and sticks of tricalcium phosphate (TCP, Biosorb, SBM, Lourdes, France). All patients were seen at three, six and twelve months, then once a year for at least four years with clinical and radiological evaluation at each visit. At the final follow up visit, no radiologic signs of pseudoarthrosis were found in either group with a minimum follow-up of 4 years. The appearance of bone callus was considered satisfactory at 6 months in all cases; moreover callus seemed to be more important in the TCP series, although this assessment was subjective. TCP resorption was total after 2 years, while allograft fragments were visible on x rays after 2 years. Minor mechanical complications occurred but did not influence the results. Loss of correction was 8% of that initially obtained in the allograft group and 2% in the TCP group. Loss of correction did not progress after 6 months in the TCP group and after 2 years in the allograft group. Based upon this experience, the use of synthetic bone substitutes such as TCP would appear to be a valuable alternative to allografts in posterolateral spinal arthrodesis for idiopathic scoliosis, and it would eliminate the risk of viral contamination inherent to allograft implantation. To our knowledge, there have been no previous comparative studies concerning the use of tricalcium phosphate versus allograft in the literature. PMID- 9415730 TI - Escalation of automutilation of the hand. PMID- 9415731 TI - Synovial osteochondromatosis: an unusual cause for subacromial impingement. AB - Impingement syndrome is usually caused by encroachment on the rotator cuff of the acromion, coracoacromial ligament, acromioclavicular joint or coracoid process. Bursal causes of impingement are rare but include rheumatoid thickening, and thickening from previous and iatrogenic causes such as sutures, pins, or wires left from previous surgery. We report a case of synovial osteochondromatosis presenting as chronic impingement syndrome. Synovial osteochondromatosis may be difficult to differentiate from chronic calcific tendinitis, but we describe a radiological sign and its anatomical basis that can differentiate between the two. If densities are seen on plain radiographs distal to the greater tuberosity, then loose bodies within the subacromial bursa should be considered. PMID- 9415732 TI - Changes in spectral measures and voice-onset time with age: a cross-sectional and a longitudinal study. AB - In this study, we searched for changes in spectral features (during the sustained vowels [a] and [i]) and the voice onset time (VOT, of [pa] and [ka]) of the ageing voice. A longitudinal study (on 20 participants) and a cross-sectional study (on 265 participants) were conducted. The spectral parameters under consideration were: the difference in amplitude between the first and the second harmonic, the energy difference between the frequency band below 1 kHz and 2-4 kHz, and the energy difference between < 1 kHz and 4-5 kHz. The VOT was also measured. Comparing five age bands of men and women, the cross-sectional study revealed a stronger H2 relative to H1 for women above 60 years pronouncing both vowels [a] and [i]. A relatively stronger spectral level was found between 4 and 5 kHz in elderly men and women (60+) for [i] compared to young men and women (20 29 years). Significant differences in the longitudinal study were not identical to those of the cross-sectional study. After a time interval of 30 years, 20 men produced a relatively weaker 2- to 4-kHz part of the spectrum and the VOT became much longer. It was impossible to attribute the changes purely to the process of ageing. Situations and emotions also play an important role and contribute to the different findings of the two groups of participants under consideration. PMID- 9415733 TI - Measuring voice under teachers' working circumstances: F0 and perturbation features in maximally sustained phonation. AB - This study is part of a larger project, the aim of which is to develop a method for collecting voice data in working places. In this part, maximally sustained phonation was studied to assess its stability in this kind of uncontrolled circumstances. The subjects were 11 female schoolteachers. Different portions of a sustained /a/ and two consecutively produced samples were measured. The acoustic variables measured were fundamental frequency (F0), jitter and shimmer. The results showed that two consecutively produced phonations were quite similar, but the place of the sample within the sustained vowel had a very small but statistically significant effect on the values of F0 and shimmer. Jitter varied quite a lot, and could perhaps be considered to be dependent on the recording circumstances. PMID- 9415734 TI - Acoustic measures of dysphonic severity across and within voice types. AB - The purpose of the study was to explore an interaction between pathologic voice type and the acoustic prediction of dysphonic severity. One hundred and two phonatory samples, representing a wide range of laryngeal conditions, were categorized by listeners into three voice types: breathy, rough, and hoarse. A second group of trained listeners rated the severity of the samples on a 7-point scale. Twenty-five frequency- and time-domain measures were used to predict perceptual severity. Multiple regression analyses showed that the most useful measure for the prediction of severity across voice types was noise-to-harmonic ratio (NHR). The severity of individual voice types was predicted with differential results on the basis of both short- and long-term measures of perturbation. PMID- 9415735 TI - Effects of transglottal pressure change on fundamental frequency of phonation: preliminary evaluation of the effect of intraoral pressure change. AB - The effect of transglottal pressure on the fundamental frequency of phonation was investigated in 14 subjects (12 adults and 2 children). A sudden change in intra oral pressure was produced during sustained phonation. The effects of this pressure change were not monotonic, even within the modal register (around the upper end of the speaking range). The physical properties of the vocal folds related to the cricothyroid and thyroarytenoid muscles seemed to be important. PMID- 9415736 TI - Effect of cochlear implantation on nasality in post-lingually deafened adults. AB - The present study addresses the effect of cochlear implantation on nasality in 21 post-lingually deafened Dutch subjects. All subjects received the Nucleus 22 implant (MSP version). Speech recordings were made pre-implantation and 3 and 12 months post-implantation with the implant switched on and off. Nasality measurements were performed on a standard text and on two sentences without nasal phonemes. The results show that post-lingual deafness in individuals can result in a deviant degree of nasality in speech production. However, the nasalance value of 86% of the subjects of our study fell within the normative range defined as the mean +/- 2 standard deviations of the normal population. After implantation we found no statistically significant effect of implant use. However, individual nasality values outside the normative range may improve. Furthermore, 12 months post-implantation we found a significant decrease in the variability of the nasalance values obtained for two sentences without nasal phonemes. PMID- 9415737 TI - Investigation of buffalopox outbreaks in Maharashtra State during 1992-1996. AB - During 1992-96, outbreaks of buffalopox zoonosis were reported from different villages in Jalgaon, Dhule and Beed districts of Maharashtra State. In humans, pox lesions were observed on the hands whereas in affected buffaloes and cows the lesions were noticed mainly on the teats and udder. Twenty two virus strains were isolated from the skin scabs collected from infected humans and milch animals. Neutralizing antibodies were detected not only in the sera of affected humans but also in their contacts. Detection of antibodies in young individuals from endemic area, who were neither vaccinated for smallpox nor had any contact with buffaloes or history of any poxvirus disease, is suggestive of occurrence of subclinical infection. A few children who had no contact with infected animals also showed clinical manifestations with disseminated lesions on the face, arm and buttocks, and thus suspected to have acquired infection through their infected parents or other family members indicating a possible man to man transmission. Therefore, in the light of discontinuation of smallpox vaccination, buffalopox outbreaks need to be monitored carefully as this may emerge as a serious zoonotic disease in India. PMID- 9415738 TI - Restriction fragment length polymorphism of Mycobacterium tuberculosis strains from various regions of India, using direct repeat probe. AB - Intraspecies differentiation was studied on 68 M. tuberculosis strains obtained from 6 states of India by restriction fragment length polymorphism (RFLP) using a direct repeat probe (DR probe) hybridised with Alu I digest of DNA. Most strains showed polymorphism based patterns that comprised between 2 to 7 bands and were grouped into 26 RFLP types. Of the 11 strains tested from Amritsar, 8 were RFLP type 5; the remaining 3 were of type 11 and were exclusively confined to this region. The strains from other regions were more heterogeneous. We confirm that DR-associated RFLP can be an excellent tool for the differentiation of M. tuberculosis strains. Depending on their geographical origin, these strains can be differentiated to a large extent by DR fingerprinting. PMID- 9415739 TI - Investigation of the outbreak of cholera in Alleppey & Palghat districts, south India. AB - In May 1996, a massive outbreak of cholera occurred in Alleppey district of Kerala which spread to Palghat district by July 1996. Of the 575 patients hospitalized at the Alleppey Medical College hospital between May 1 and August 2, 1996, 30 deaths occurred with a case fatality rate of 5.2 per cent while of the 638 diarrhoea patients admitted at Agali PHC of Attapadi area in Palghat district, 30 (4.7%) deaths were recorded. Clinically, the patients had profuse watery diarrhoea with vomiting. The epidemic of cholera in Alleppey and Palghat districts was caused by V. cholerae O1 of the EITor biotype, Ogawa serotype which possessed both the ctxA and tcpA genes when examined by multiplex PCR. Gross contamination of water sources was incriminated as the cause of the epidemic. PMID- 9415740 TI - Environmental isolation of Cryptococcus neoformans var. neoformans from Vellore. AB - We have isolated C. neoformans from various environmental niches, including our hospital environment. Twenty one samples of weathered pigeon droppings and 86 samples of Eucalyptus bark and debris were collected from various sites around the hospital and college. C. neoformans var. neoformans was isolated from 14 of the 21 (67%) samples of weathered droppings of pigeons collected from the ledges of windows of wards and departmental buildings in the hospital. C. neoformans was not isolated from any of the Eucalyptus samples. These findings document for the first time the presence of C. neoformans var. neoformans in the hospital environment. Although ubiquitous in the environment, its presence in hospital surroundings stresses the need for systematic cleaning of sites inhabited by pigeons. However, we failed to isolate C. neoformans from the limited number of Eucalyptus samples examined. PMID- 9415741 TI - Norfloxacin induced resistance to fluoroquinolones & structurally unrelated antimicrobial agents in coagulase negative staphylococci. AB - Nine clinical isolates of coagulase negative staphylococci (CONS) susceptible to norfloxacin (MIC 1.8-2 micrograms/ml) were manipulated in vitro to induce norfloxacin resistance by means of serial passage in brain heart infusion broth containing increasing concentrations of norfloxacin. Exposure of CONS to norfloxacin resulted in 18 to 20 times increase in MIC of norfloxacin and change in in vitro susceptibility to ciprofloxacin, pefloxacin, ofloxacin, kanamycin, neomycin and tobramycin, indicating development of cross resistance to fluoroquinolones and aminoglycosides. These results show that exposure to increasing concentrations of norfloxacin can induce the development of resistance to various antimicrobial agents, suggesting its mutagenic role. PMID- 9415742 TI - An outbreak of food poisoning in Tamil Nadu associated with Yersinia enterocolitica. AB - An outbreak of food poisoning in a Tamil Nadu village, affecting 25 of 48 individuals who participated in a feast, was investigated. The risk of developing illness was associated with consumption of buttermilk (relative risk 3.8). None of the food items consumed during the feast was available for analysis. Toxin producing Y. enterocolitica (serotype 3, biotype 4) was grown from 1 of 11 stool samples from affected individuals, as well as from a water sample from the source used to dilute the buttermilk. High titres of antibody of Yersinia were detected in 2 of 12 patients but in neither of the two groups of controls. Toxin production was noted in buttermilk incubated for 6 h with Y. enterocolitica. This is the first report from India of a food poisoning outbreak associated with this organism. PMID- 9415743 TI - Serum & muscle magnesium in Indians with cirrhosis of liver. AB - Magnesium status of Indian patients with cirrhosis of liver (alcoholic and non alcoholic) and the role of low magnesium in neuromuscular and neuropsychiatric manifestations of chronic liver disease were evaluated in 76 male cirrhotics (alcoholic 37, aged 48 +/- 11 yr, non alcoholic 39, aged 47 +/- 12 yr) and 37 male controls (aged 49 +/- 11 yr). Serum magnesium levels were similar in the 3 groups studied. Muscle magnesium in both groups of cirrhotics were significantly lower than in controls (alcoholic cirrhosis 33.77 +/- 16.85; non alcoholic cirrhosis 37.93 +/- 18.86 and controls 70.52 +/- 6.49 mEq/kg fat free dry mass; P < 0.001). Multiple regression analysis comparing muscle magnesium with clinical and biochemical parameters in cirrhosis showed that hepatic encephalopathy was associated significantly and independently with low muscle magnesium (Beta = 0.313; P = 0.01). These results indicate that patients with cirrhosis have significantly lower muscle magnesium than controls and suggests that low muscle magnesium may be a factor associated with or precipitating hepatic encephalopathy. PMID- 9415744 TI - Reactive oxygen species formation in peripheral blood neutrophils in different types of smokers. AB - The present study was envisaged to assess the state of oxidative metabolism of neutrophils, recovered from smokers (cigarette, beedi, hookah and mixed products) and non-smokers. Superoxide anion (O2.-) production was significantly higher in neutrophils from all groups of smokers (P < 0.001). Total leukocyte count (TLC) was significantly more in symptomatic subjects among the cigarette, hookah and mixed smokers (P < 0.05). Total neutrophil count (TNC) was significantly higher in symptomatic than asymptomatic subjects of hookah and mixed smoking groups (P < 0.05). In the pathogenesis of chronic obstructive pulmonary disease (COPD) in tobacco smokers, the role of leukocytosis, increased neutrophil sequestration into the lung, increased neutrophil toxic oxygen species including superoxide release in the lung may cause direct injury to lung tissues. PMID- 9415745 TI - Effects of vanadyl sulphate on glucose homeostasis in severe diabetes induced by streptozotocin in rats. AB - Rats made severely diabetic by an i.v. injection of 50-55 mg/kg streptozotocin (STZ), 15 days later showed blood glucose > 500 mg/dl, quadrupled daily water intake and plasma insulin of 14 +/- 3 microU/ml, about 25 per cent that of normal rats. Subsequently, these rats in two groups, received 0.5-1 mg/ml vanadyl sulphate in base solution (vanadyl) orally (Group I) and base solution containing 50 mEq/1 NaCl (Group II). Since 90 days of vanadyl therapy could not decrease blood glucose of group I (420 +/- 10 mg/dl) to normal levels, euglycaemia was achieved for a period of two months by intraperitoneal (i.p.) injection of NPH insulin. The required daily doses of insulin in vanadyl-treated rats of group I (8 +/- 1 U/kg/day) were only 8 per cent of those in group II animals (103 +/- 7 U/kg/day). In conclusion, it seems that vanadyl per se cannot induce normoglycaemia in diabetic rats with very low plasma insulin levels, but can augment the sensitivity of peripheral tissues to insulin. PMID- 9415746 TI - The increased formation of alkali-labile sites by a spin-trapping agent alpha phenyl-N-tert-butylnitrone in gamma-irradiated frozen aqueous solution of DNA. PMID- 9415747 TI - Changes in Serum Ceruloplasmin activity after whole body irradiation of mammals. AB - Chronological changes in ceruloplasmin oxidase activity after gamma-irradiation with semilethal doses of 3-5 Gy, were investigated in four mammalian species; rat, guinea pig, lamb and pig. The ceruloplasmin activity increased soon after irradiation but later decreased. Although the extent of the increase and its time course varied among species, it was most remarkable in rats and least so in guinea pigs, with the highest activity generally attained at 12 h after irradiation. In contrast, erythrocyte and leukocyte counts decreased after irradiation, and showed minimum values when ceruloplasmin levels were maximum. These results suggest that ceruloplasmin is involved in the recovery from radiation disease. PMID- 9415748 TI - Mutational specificity of the ferrous ion in a supF gene of endonuclease III/VIII deficient Escherichia coli. AB - When 125 microM Fe2+/EDTA treated plasmid pUB3 was used to transfect an Escherichia coli NKJ2004 (nth nei) host, which is totally defective in glycosylases for thymine glycol and 5-hydroxycytosine, a 3.7 fold increase in mutation frequency was observed. Among 46 supF mutants sequenced, 28 had base substitutions, with G:C-->C:G transversion predominant (14 cases), followed by G:C-->T:A transversion (6 cases) and G:C-->A:T transition (6 cases). The results are consistent with our previous Fe2+ mutagenesis results where, in the wild type host, 78% were base substitutions, with G:C-->C:G transversion (59%) predominant, followed by G:C-->T:A transversion (28%) and G:C-->A:T transition (11%). Treatment of pUB3 DNA with Fe2+/EDTA did not yield formation of Endonuclease III sensitive sites. The possibility of 5-hydroxycytosine as the causative lesion for Fe2+ induced G:C-->C:G transversion is discussed. PMID- 9415749 TI - Alkaline denaturation of dentin--A simple way to isolate human tooth enamel for electron spin resonance dosimetry. AB - Electron spin resonance (ESR) of tooth enamel is a recently developed method for the retrospective dose estimation of human radiation exposures. The assay requires isolation of enamel from dentin, which is difficult because the boundary between enamel and dentin is not easily discernible. Here we describe a simple method for isolating enamel by alkaline denaturation of dentin. The method requires 4 weeks, but scratching of the denatured and hence softened dentin is needed only once a week. Above all, no special skill is required. We found that the alkaline treatment did not cause deterioration of the ESR signal recorded in enamel exposed to 2 Gy of gamma rays prior to its isolation. The assay is particularly suited for teeth containing many cracks that were generated during long-term storage after extraction of the teeth. Such teeth tend to disintegrate during enamel isolation processes, which poses difficulties to isolate enamel mechanically from individual small pieces. PMID- 9415750 TI - p53-dependent signal transduction induced by stress. AB - While recent advances in elucidating the enzyme/substrate relationship of phosphorylation cascades have demonstrated several distinct pathways in membrane cytoplasm signaling, the molecular dissection of p53 and the products of other proto-oncogenes have greatly promoted the studies of nuclear signaling and expanded the checkpoint concept to mammalian cells. The growing list of p53 activating factors ranges from genotoxic agents to non-genotoxic stresses. The diverse involvement of p53 and its close linkage to other nuclear and extranuclear signaling networks force us to reconsider the concept of cellular stress response. A signaling network emerges from crosstalks between different types of stress in the same signaling pathway and crosstalks between different pathways in response to the same stress. We review the present knowledge on cellular stress signaling with emphasis on the crosstalks between different pathways and the molecules which mediate these crosstalks and offer our concept of signaling checkpoints. The importance of stress signaling checkpoints in cancer evolution and cancer therapy is also discussed. PMID- 9415751 TI - The Pledge Allegiance Generation. PMID- 9415752 TI - GP vs. specialist. PMID- 9415753 TI - More about specialists and GPs. PMID- 9415754 TI - Fluoride. PMID- 9415756 TI - Bacterial endocarditis. PMID- 9415755 TI - Tooth decay rising? PMID- 9415757 TI - Hygienists and dentists. PMID- 9415758 TI - Today's impatient patient. AB - Fee-for-service dentists face the daily challenge of differentiating themselves from discount providers while delivering quality care to patients pressed for time. Some thoughts on how to make the best of limited time. PMID- 9415759 TI - Are you operating at OPC? AB - You don't need an M.B.A. to know that to thrive in today's competitive market, you need to run your practice like a business. It's a matter of keeping productivity high, while holding the line on costs and stress. PMID- 9415760 TI - The challenge of change. AB - To succeed in modern life, you must learn to be comfortable with the inevitability of rapid change. The author presents five keys to the successful management of change. PMID- 9415761 TI - Get with the Net. AB - The Internet is not some passing fancy that you can safely ignore. It is rapidly becoming the way of doing business in a high-tech world. Don't get left behind. PMID- 9415762 TI - Marketing with a patient focus. AB - Marketing is not a dirty word. It's a process that enables you to better understand the needs and wants of your patients. It's about listening and learning from your patients as a way to improve the care you provide. PMID- 9415763 TI - Reduction of postoperative pain: a double-blind, randomized clinical trial. AB - The authors investigated the capability of two commonly used basing techniques to reduce postoperative sensitivity. The authors measured the time it took the subjects to respond to a standardized stimulus of cold water (cold response measure, or CRM) at baseline and one week after treatment. The authors found no significant reduction in the mean CRM for the group receiving Amalgambond Plus (Parkell), but they did find a significant decrease in the mean CRM for the group receiving Copalite (Harry J. Bosworth Co.) with or without Vitrebond (3M). The authors concluded that the subjects in the Amalgambond Plus group experienced no more sensitivity to cold at one week postoperative than they did at baseline, while the subjects in the Copalite/Vitrebond group did. PMID- 9415764 TI - Tobacco control activities in U.S. dental practices. AB - The authors surveyed general dentists, periodontists, pediatric dentists and dental hygienists in three U.S. geographical regions to estimate the percentage who practiced tobacco use cessation activities in their dental offices. A total of 1,746 dentists and 723 dental hygienists completed either a long or short version of a mailed survey or telephone interview. The authors found that tobacco cessation activities are not a routine part of dental practice and that tobacco control activities and training vary by dentist type and geographical region. PMID- 9415765 TI - Diagnosing depression in patients with chronic facial pain. AB - Depression is a common serious disorder that has a high comorbidity with chronic facial pain. This article provides an overview of the clinical presentation, pathophysiology and diagnosis of depression. In particular, it addresses some of the problems associated with identifying depressive symptoms in dental patients who have chronic facial pain. PMID- 9415766 TI - Special Olympics, special smiles: assessing the feasibility of epidemiologic data collection. AB - No comprehensive national study has ever been completed on the oral health status of people with disabilities, their patterns of use of oral health services and access-to-care barriers. The authors describe the Special Olympics, Special Smiles program, conducted as part of the New Jersey Summer Special Olympics Games, and assess a pilot-tested model for collecting epidemiologic data. The results of this initial data collection are also compared with the goals of the U.S. Public Health Service, as outlined in the Healthy People 2000 publication. PMID- 9415767 TI - Reflections on taste for oral health professionals. PMID- 9415768 TI - Developing a great dental team. PMID- 9415769 TI - Use of rotational movements to remove mandibular molars. PMID- 9415770 TI - Facilitating the removal of tissue conditioner from the denture intaglio. PMID- 9415771 TI - Demographics and practice characteristics of dentists participating and not participating in managed care plans. AB - This article is the first in a series compiling the results of the 1995 Managed Care Marketplace Information Survey conducted by the ADA Council on Dental Practice. It addresses the demographic and practice characteristics of dentists participating and not participating in managed care plans. Dentists with at least one contract were somewhat more likely to be located in the Pacific or Middle Atlantic regions, be female and have practiced less than 10 years. Dentists who participated in at least one managed care contract worked slightly more hours per week, saw slightly more total patients per week, saw more emergency and walk-in patients and had shorter appointments. PMID- 9415772 TI - Teledentistry: legal and regulatory issues explored. PMID- 9415773 TI - Angina pectoris from an unexpected cause. AB - A 53 year-old-man developed classical angina pectoris, presumably the result of coronary atherosclerosis. Thirty-two years before, an abnormality on a chest film had been described as a "prominence of the upper portion of the left cardiac border." The "prominence" was actually an angiosarcoma, which eventually compromised blood flow in the left coronary system. PMID- 9415774 TI - Sonohysterography: a unique technique for the evaluation of gynecologic disorders. PMID- 9415776 TI - HMO figures more troubling news for poor. Not enough of budgeted money appears to be filtering down to patient care. PMID- 9415775 TI - Inappropriate use of DEA numbers. PMID- 9415777 TI - Salvaging life from the grim reaper. PMID- 9415778 TI - Physicians, PAs and the facts. PMID- 9415780 TI - Physicians at risk: profiling. PMID- 9415779 TI - Coding concepts. PMID- 9415781 TI - Foundation for Medical Care. PMID- 9415782 TI - Opportunities and challenges in educating community-responsive physicians. PMID- 9415783 TI - Latino health beliefs and locus of control: implications for primary care and public health practitioners. PMID- 9415784 TI - Why a peer intervention program for Mexican-American women failed to modify the secular trend in cancer screening. AB - INTRODUCTION: We evaluated an intervention program for Mexican-American women to increase Pap smear and mammography screening. METHODS: The three-year intervention included the presentation of role models in the media and reinforcement by peer volunteers. We used a two-community (intervention and comparison) pre-post test design. Activities were targeted to a mainly Spanish speaking, poverty-level, immigrant population. Pre- and postintervention screening rates were based on independent random samples of Mexican-American women 40 years and older. RESULTS: Women reported a 6% absolute increase in Pap smear use similar to the 7% increase in the comparison community. Both communities experienced large but similar increases in recent mammography use (17% and 19%). Adjusting for differences in demographic factors, intervention and comparison changes remained identical. CONCLUSIONS: Our peer intervention failed to accelerate the secular trend in cancer screening low-income Mexican-American women. Likely, promotional activities were too diffuse and the comparison community was contaminated with similar interventions. Strong social and market forces make it difficult to measure the effect of a specialized intervention on cancer screening rates. PMID- 9415785 TI - The influence of fatalism on self-reported use of Papanicolaou smears. AB - OBJECTIVE: Our objective was to examine the demographic and other predictors of fatalistic beliefs among Latinas (Hispanic women) and Anglo (non-Hispanic Caucasian) women and to assess the impact of these beliefs on the use of cervical cancer screening services. METHODS: We used ethnographic interviews and a cross sectional telephone survey in Orange County, California. Our sample included 94 Latinas and 27 Anglo women selected through organization-based network sampling for the ethnographic interviews and 803 Latinas and 422 Anglo women randomly selected for the telephone survey. RESULTS: Latina immigrants (Latinas born outside the United States) were more likely than U.S.-born Latinas or Anglo women to have fatalistic beliefs. Immigration, education levels, and insurance status predicted fatalistic beliefs. Fatalistic beliefs were independent predictors of Pap smear use by Latinas but not Anglo women. For example, after adjusting for potentially confounding variables, Latinas who believed that fate was a risk factor for cervical cancer (odds ratio [OR] = .58), that they would rather not know if they had the disease (OR = .58), and that there is nothing one can do to prevent it (OR = .45) were less likely than others to report that they have had a Pap smear within the prior three years. Health insurance status, marital status, and immigration also predicted use of Pap smears. Insured Latinas were more likely than uninsured Latinas (OR = 2.89) to report having a Pap smear within the prior three years. In addition, married Latinas (OR = 2.32) and Anglo women (OR = 3.09) were more likely than unmarried women to report having appropriate cervical cancer screening. Finally, Latina immigrants were less likely than other Latinas to report having a Pap smear (OR = .26). CONCLUSIONS: We conclude that fatalistic beliefs are among the factors that negatively influence Latinas' use of Pap smears and that it is important for health care professionals to address those beliefs. Continued efforts are also necessary to decrease the economic and structural barriers to cervical cancer screening. PMID- 9415786 TI - Pap smear outreach: a randomized controlled trial in an HMO. PMID- 9415787 TI - Recall and treatment decisions of primary care providers in response to Pap smear reports. AB - INTRODUCTION: Although the frequency of cervical cancer screening has been extensively studied, little is known about how clinicians decide to screen or recall patients for Pap smears. This study reports the management decisions made by office-based clinicians for 10 different Pap smear reports describing adequacy limitations and cytological diagnoses. METHODS: We surveyed 186 clinicians using a commercial laboratory in the southeastern United States and analyzed results by frequency and comparison statistics. RESULTS: Our respondents were 148 clinicians (79.6% response rate) from different specialties. There was variation in reported management of inflammation, atypia, and low-grade abnormalities (LGSIL), in regard to recall for repeat or routine testing as well as arranging colposcopy. In only 3 of 10 Pap smear results did more than 50% of respondents agree on a specific test recall interval. CONCLUSIONS: The variation in responses from office-based clinicians suggests either uncertainty or different opinions in making recall and treatment decisions for smears of limited quality even when associated with cytologic abnormalities. These differences may have relevance to outcomes, clinician workload, and costs of care in cervical cancer screening. PMID- 9415788 TI - Physicians' reactions to change in recommendations for mammography screening. AB - INTRODUCTION: In December 1993 the National Cancer Institute (NCI) decided to replace its mammography screening guidelines with a Statement of Evidence on Breast Cancer Screening. The Statement of Evidence represented a departure from the institute's previous policy of recommending routine mammography screening in women 40-49 and annual screening in women 50 and over. This study assesses knowledge of and attitudes toward the Statement of Evidence among primary care physicians. In addition, we explore the extent to which physicians changed their individual clinical policies on mammography screening in response to the Statement of Evidence. METHODS: Between October 1994 and June 1995, 545 randomly selected North Carolina primary care physicians completed a mailed questionnaire (overall survey response rate = 42%). RESULTS: Awareness of the Statement of Evidence was high (83%), but attitudes toward it were negative, with a majority of physicians stating that the change in policy was confusing to women and physicians. About 8% of physicians who were aware of the Statement of Evidence changed their practice accordingly. Most physicians reported recommending routine mammography screening in women 40-49 and annual screening in women 50 and over. A majority stated they believe scientific evidence supports these practices. CONCLUSIONS: When faced with a new policy in which guidelines are no longer provided and evidence supporting less use of an established technology is conveyed, physicians demonstrated disinclination to change. The impact of changes in recommendations on physician practice is an important consideration for those who develop and promote screening policies. PMID- 9415789 TI - Beliefs and mammography screening. AB - INTRODUCTION: Breast cancer is a leading form of preventable cancer among women in the United States. Despite improvements in mammography and other early detection techniques, special populations, including older and minority women, continue to experience high incidence and mortality rates. Knowledge, attitudes, and beliefs are ubiquitous constructs in preventive medicine, health behavior, and behavioral epidemiology. These constructs often are used to explain variation in health screening behavior. While all three have been examined in relation to mammography screening, concentration on the single category of beliefs and the relation between specific beliefs and mammography screening practices has remained largely uninvestigated. METHODS: Using logistic regression modeling, we examined the relationship between four individual beliefs and mammography screening in a cross-sectional study of 407 women. RESULTS: After we controlled for confounding factors in a multivariable analysis, belief in the efficacy of early detection in improving breast cancer outcome (odds ratio [OR] = 2.98; 95% confidence intervals [CI] = 1.62, 5.47) and perceived risk (OR = 0.49; 95% CI = 0.26, 0.94) were significantly associated with screening practice. Belief that mammography is dangerous (OR = 0.46; 95% CI = 0.18, 1.18) or painful (OR = 1.25; 95% CI = 0.75, 2.08) was not significantly associated with screening practice. CONCLUSIONS: Information on the relationship between beliefs and screening practices may be used both to understand screening behaviors and to develop targeted strategies to improve mammography compliance. PMID- 9415790 TI - Screening asymptomatic women for ovarian cancer: American College of Preventive Medicine practice policy. PMID- 9415791 TI - Modeling fracture risk using bone density, age, and years since menopause. AB - INTRODUCTION: Preventive strategies are essential for reducing the incidence of osteoporosis and its consequences. However, simple algorithms that predict an individual's future risk of fractures are scarce. The purpose of the study was to define a clinical decision aid that enables physicians to project an individual's life-time fracture risk and therefore institute preventive therapies. METHODS: A predictor equation for bone loss was developed using bone mineral density (BMD), age, years since menopause, and weight. This was applied to normal and osteoporotic women, ages 40-80 years (n = 117) screened for osteoporosis studies. RESULTS: A spinal BMD cutoff of 0.86 gm/cm2 had a sensitivity of 90% and a specificity of 60% for detecting subjects with vertebral fractures and was therefore defined as a high-risk BMD. Using the parameter estimates from the above equation and an individual's clinical data, we derived prediction curves to forecast the age at which that individual would reach the above defined high-risk BMD, and therefore that person's expected number of remaining life-years at high risk for fractures. CONCLUSIONS: This study proposes a conceptual framework for the development of a clinical decision aid to provide guidelines for the prevention of osteoporosis. A longitudinal study that incorporates other variables such as prevalent fractures and biochemical markers of bone turnover would further validate this model and enhance its application. PMID- 9415792 TI - The effect of health insurance coverage on the appropriate use of recommended clinical preventive services. AB - INTRODUCTION: Lack of health insurance coverage has been shown to reduce use of some preventive services. However, even when care is free or fully covered by insurance, clinical preventive services are not used at recommended levels. This study investigates the impact of different levels of health insurance coverage (ranging from none, some, most, and all preventive services covered) on the use of recommended clinical preventive services for adult men and women. METHODS: Logistic regression was used to estimate the effect of different levels of health insurance coverage for preventive care on the probability of receiving six different clinical preventive services including periodic health exam, blood pressure screening, cholesterol screening, Pap smear, clinical breast exam, and screening mammography, as well as all recommended services for a given age and gender group. The study sample of adults ages 18 to 64 is from the Centers for Disease Control's 1991 Behavioral Risk Factor Surveillance System (BRFSS) (n = 53,981). RESULTS: The results demonstrate a positive and statistically significant dose-response relationship between level of health insurance coverage for preventive care and receipt of recommended preventive services in adult men and women. The odds ratios (ORs) of men who had full coverage for preventive care receiving recommended preventive services compared to men with no coverage for preventive care ranged from 1.8 to 2.8. For women the ORs were 1.2 to 2.0. The ORs for men with "most" preventive services covered compared to none covered ranged from 1.3 to 2.1, and for women from 1.2 to 2.0. CONCLUSIONS: The level of health insurance coverage for preventive care is one of the most important determinants of receipt of recommended preventive services for adult men and women 18-64 years of age. These results suggest that comprehensive health insurance coverage for clinical preventive care may significantly increase receipt of recommended preventive services for this population. PMID- 9415793 TI - The impact of an elementary school-based violence prevention program on visits to the school nurse. AB - INTRODUCTION: In Tucson, Arizona, an elementary school-based violence prevention program (PeaceBuilders) was implemented during the 1994-1995 school year. Anecdotal evidence from school nurses suggested that children were visiting the nurse less often following the implementation of the program. We examined nurses' logs to assess whether the program had an impact on visits to the school nurse. METHODS: For the school years 1993-1994 and 1994-1995, the weekly number of nurse visits for all reasons, all injuries, and injuries caused by fights in each of the four PeaceBuilders schools were compared with those for three control schools. As part of a planned evaluation, schools had been matched on demographic factors and randomly assigned as intervention or control schools. RESULTS: Between 1993-1994 and 1994-1995, the rate of visits/1,000 student days decreased 12.6% in the intervention schools while remaining unchanged in the comparison schools. The same trend was detected for injury-related visits. Rates of fighting related injuries changed little in the intervention schools but increased 56.0% in the control schools. An analysis of covariance confirmed that injuries and visits to nurses decreased in intervention schools relative to control schools. CONCLUSIONS: These data indicate that in the intervention schools, injuries and visits to the school nurse decreased over the two-year period and that the intervention may have contributed to this change. They also suggest that visits to the school nurses' office may be a useful tool to evaluate some types of elementary school-based violence prevention programs. PMID- 9415794 TI - Brief intervention in a primary care setting for hazardous drinkers. AB - INTRODUCTION: The study was designed to test a brief intervention for reducing alcohol consumption among moderate to heavy (hazardous) drinkers in a busy HMO primary care setting. METHODS: In a randomized controlled trial, hazardous drinkers (n = 516) were identified by the AUDIT screening questionnaire. Intervention included brief clinician advice (30 seconds), a 15-minute motivational session by counselors, and printed materials. RESULTS: At six-month follow-up, intervention subjects reported fewer total standard drinks in the past three months (176 versus 216, P = .04, one-tailed) and fewer drinking days per week (2.8 versus 3.3, P = .02) than controls, but similar drinks per drinking day (3.3 versus 3.5; P = .13). At 12 months, intervention subjects again reported fewer drinking days per week (2.7 versus 3.1; P = .04) than controls, but similar numbers of standard drinks (157 versus 179; P = .13) and drinks per drinking day (3.6 versus 3.3; P = .20). Intervention subjects were somewhat more likely than controls to report drinking within daily recommended limits (< or = 3 for men, < or = 2 for women) at both six months (79% versus 71%; P = .06) and 12 months (80% versus 73%; P = .07), but did not differ significantly from controls on other drinking outcomes (percent abstinent, frequency of drinking > or = 6 drinks per drinking occasion, estimated peak blood alcohol concentration), or use of medical care in the year following intervention. CONCLUSIONS: A one-time, brief motivational intervention using minimal clinician time supplemented by trained counselors resulted in a modest reduction in frequency of alcohol consumption in a busy primary care population. Future research should focus on strengthening and maintaining intervention effects. PMID- 9415795 TI - Sexually assaulted males: 115 men consulting a counseling service. AB - The nature of sexual assault on men and their help seeking following the assault was investigated. All men were seen at least once for face-to-face counseling at SURVIVORS, a counseling service for male victims. Data on 115 men were analyzed: 69 were assaulted while under age 16. Mean time from assault to contact with SURVIVORS was 16.4 years. 51 men (44%) were assaulted more than once. The assailant was known to the victim in all but 16 cases. 100 men (87%) were assaulted by at least one man, 7 (6%) by a man and a woman, and 8 (7%) by women. Forced anal penetration took place in 88. 27 men (23%) feared for their lives during the assault. 88 men (79%) sought no help and only 17 men (15%) reported to police. For victims assaulted under the age of 16, the assault was more likely to be their first sexual experience and they were more likely to delay contact with SURVIVORS for more than 17 years. They were also less likely to report to police. Victims assaulted by more than one person were more likely to have been assaulted by strangers, by women, and to have suffered physical harm. They were less likely to have experienced the assault as their first sexual experience. PMID- 9415797 TI - The effects of anxiety and distraction on sexual arousal in a nonclinical sample of heterosexual women. AB - The effects of anxiety and distraction on sexual arousal in a nonclinical sample of heterosexual women between the ages of 19 and 35 were studied. Using a dichotic listening paradigm, the study extended Geer and Fuhr's (1976) research by examining the effects of distraction on sexual arousal in women. Results indicated that both vaginal pulse amplitude and subjective measures of sexual arousal vary as a function of distraction level, with increased distraction leading to decreased arousal. However, the data failed to support Masters and Johnson's (1970) assertion that anxiety decreases sexual arousal. Although no significant effect for anxiety emerged using a physiological measure of sexual arousal, a significant Anxiety x Distraction interaction was observed using a subjective measure of sexual arousal. Several competing interpretations of this interaction are discussed. PMID- 9415796 TI - Transvestism: a survey of 1032 cross-dressers. AB - One thousand and thirty-two male periodic cross-dressers (transvestites) responded to an anonymous survey patterned after Prince and Bentler's (1972) report. With few exceptions, the findings are closely related to the 1972 survey results. Eighty-seven percent described themselves as heterosexual. All except 17% had married and 60% were married at the time of this survey. Topics surveyed included demographic, childhood, and family variables, sexual orientation and sexual behavior, cross-gender identity, cross-gender role behavior, future plans to live entirely as a woman, and utilization of counseling or mental health services. Of the present sample, 45% reported seeking counseling compared to 24% of the 1972 survey, and those reporting strong transsexual inclinations were up by 5%. Today's transvestites strongly prefer both their masculine and feminine selves equally. A second research objective was to identify variables discriminating between so-called Nuclear (stable, periodic cross-dressers) and Marginal transvestites (more transgendered or transsexually inclined); 10 strongly discriminating parameters were found. The most important are (i) cross gender identity, (ii) commitment to live entirely as a woman, (iii) taking steps toward body feminization, (iv) low sexual arousal to cross-dressing. Neither age nor experience as a cross-dresser were found to be correlates of cross-gender identity. Although the present generation of transvestites describe themselves much as did similar subjects 20 years ago, the percentage migrating toward full time living as a woman is greater. PMID- 9415798 TI - Social class background, sexual attitudes, and sexual behavior in a heterosexual undergraduate sample. AB - To further the understanding of the relationship between social class and sexual attitudes and behavior, we present data from a study of undergraduate students. We look at the education of students' fathers and how it relates to students' sexual profiles. Among the men, some traditional social class differences are found, indicating that class differences persist among some upwardly mobile men. For the women, fewer social class differences appear. Further, we compare our 1992 sample of 554 college students, 19-22 years old, with a university sample of 904 similar age students from 1967, and find our sample more coitally experienced. College students today are following norms that in the past were associated with a lower educational level. Implications of our findings for class convergence theory are addressed. Reliable birth control, gains in equality by women, and the sexual images of television and other media are discussed as major factors contributing to the increased sexual permissiveness among university students of the 1990s. PMID- 9415799 TI - Defining and measuring sexual orientation: a review. AB - How to define and identify sexual orientations for the purpose of constructing representative samples of homosexuals, bisexuals, and heterosexuals is unclear and confusing to researchers. Different definitions and measures have been proposed and used to develop samples since the 1860s when sexual orientations first gained widespread research interest. Unfortunately, the definitions and measurement tools used since then result in the selection of divergent and incomparable samples. If advances in the understanding of sexual orientations are to be made, it is critical that definitions and measures of sexual orientation be standardized. This paper reviews and critiques definitions and measures of sexual orientation that have been proposed and used by researchers over the past century. This review is intended to further our understanding of this subject and to encourage researchers to be critical of how they classify subjects based upon sexual orientation. PMID- 9415800 TI - Are transvestites necessarily heterosexual? PMID- 9415801 TI - Cloning and sequencing of the pyrG encoding cytidine 5'-triphosphate synthetase from Mycobacterium bovis BCG. AB - The pyrG gene encoding cytidine 5'-triphosphate (CTP) synthetase which catalyzes the terminal reaction for de novo synthesis of pyrimidine nucleotides was cloned and sequenced from Mycobacterium bovis BCG. The BCG pyrG was screened in lambda ZAPII library employing a DNA probe which was constructed from PCR based on a consensus pattern of glutamine amidotransferase type I. The BCG CTP synthetase deduced from a nucleotide sequence consisting of 586 amino acid residues with a calculated molecular weight of 63,642 daltons. Alignment of BCG CTP synthetase with all sequences available in GenBank indicates that the residues suggested as a catalytic triad for glutamine hydrolysis at the carboxyl terminal domain and an amino terminal segment are completely conserved among all CTP synthetases. PMID- 9415802 TI - Intracellular proteases in sporulated Bacillus thuringiensis subsp. tenebrionis: detection and analysis by gelatin zymography. AB - Three intracellular proteases were identified from sporulated cultures of Bacillus thuringiensis subsp tenebrionis by fractionation with ammonium sulfate. In this study, we detected protease activities at M(r) 92 kDa, 81 kDa and 69 kDa employing gelatin zymography. The major proteolytic activity was due to the 81 kDa protease, which was identified as a metalloprotease being inhibited by both 1,10-phenanthroline and ethylenediamine tetraacetic acid. The proteases showed maximal azocasein hydrolytic activity at 60 degrees C and were heat-activated from 40 degrees C to 60 degrees C. The 69 kDa and 81 kDa proteases were thermo inactivated at 70 degrees C and 80 degrees C respectively, while the 92 kDa protease was still active at 80 degrees C. PMID- 9415803 TI - Multiprotein complex formation on the c-myc promoter. AB - The E2F element is a cis-acting DNA sequence within the P2 promoter of c-myc proto-oncogene. While it is required for optimal transcription, the multiprotein complexes formed on this site have not been well characterized. We show that in extracts of human glioblastoma cells and NIH3T3 fibroblasts, significant E2F transcription factor binding to the c-myc E2F site occurs as a both a monomer (the active form) and as only two mutually exclusive complexes with the retinoblastoma gene product (pRb) or the cyclin A protein. The E2F protein monomer was found predominantly in the cytosolic fraction of the cellular extracts while the pRb and cyclin A complexes in the nuclear fraction, indicating that the monomer has novel physical properties. Thus, protein complex formation on the c-myc E2F site appears to contribute in a unique way to transcriptional activation. PMID- 9415805 TI - Effect of pyrrolidine dithiocarbamate on photo-induced proton transport through chloroplast membranes. AB - pH changes produced by photo-induced proton transport through chloroplast membranes in spinach were measured by a glass microelectrode. Effect of pyrrolidine dithiocarbamate on proton translocation through chloroplast membranes has been studied. Kinetic analysis of proton translocation shows that the rate is reduced as the carbamate concentration increases. The rate of proton uptake follows first-order kinetics and diminishes with increasing carbamate concentrations. The outward leakage of accumulated protons through thylakoid membranes in the dark also decreases likewise. However, the leakage of protons takes a much longer time. Pyrrolidine dithiocarbamate is an effective inhibitor of proton transport through chloroplast membranes. The results suggest that the photo-induced proton translocation is regulated by conformation change in the membrane. Higher concentration of carbamate disrupts the tertiary conformation of the membrane. The inhibition of proton transport would affect ATPase function; thus, an excess use or accumulation of pyrrolidine thiocarbamate may compromise ATP production. PMID- 9415804 TI - Identification of actin as an estradiol 17-beta-stimulated protein in the human placenta. AB - Addition of estradiol 17-beta to first trimester human placental minces resulted in an increased synthesis of a protein of apparent molecular weight 45 kDa. The specific involvement of estrogen in the stimulation of this protein was established by demonstrating a reduction in the level of this protein by the addition of CGS 16949 A, an inhibitor of aromatase, a key enzyme in the biosynthesis of estradiol 17-beta and ICI 182,780, an estrogen receptor antagonist. The protein was purified to homogeneity and N-terminal sequencing of two of the internal peptides obtained by enzymatic digestion of the protein, as well as the absence of a free N-terminal indicated that it could be actin. This was confirmed by Western blotting using commercially available actin antiserum. The role of estradiol 17-beta in the stimulation of actin synthesis in human placenta was also established by monitoring the quantitative inhibition of DNase I by actin. PMID- 9415806 TI - Differential expression of ferritin heavy chain in THP-1 cells infected with Mycobacterium bovis BCG. AB - To identify the host genes induced or suppressed by infection of mycobacteria, the reverse transcriptase polymerase chain reaction (RT-PCR) and the differential display reverse transcriptase polymerase chain reaction (DD RT-PCR) methods were used. In this study, cDNAs complement to mRNA extracted from human peripheral monocyte derived naive THP-1 cells, THP-1 cells infected with live Mycobacterium bovis BCG, THP-1 cells treated with heat-killed BCG, and THP-1 cells incubated with IgG-coated glass-beads were compared on the sequencing gel. One (TG2-1) of the clones selected by DD RT-PCR is 446 bp long and is identical to human ferritin heavy (H) chain gene. Northern blot analysis confirmed that ferritin H chain gene has been markedly over-expressed in monocytic THP-1 cells incubated with live and dead M. bovis BCG. Differential display techniques of host genes whose expression levels were varied by infection of mycobacteria could provide information about the response of macrophages to mycobacterial infection. PMID- 9415807 TI - Repetitive elements in the third intron of murine apolipoprotein A-I gene. AB - Genomic DNAs containing the gene encoding apolipoprotein A-I (apoA-I) from four strains of mice and three strains of rats have been sequenced. Some peculiar repetitive sequences were found in the third intron of apoA-I of the murine species. The striking features include regular tandem repeats of C(A)4 and C(A)6 in mice and long A-tracts in rats. Not completely identical but very similar motifs were found in mice or rats belonging to the same species while repetitive elements from different species show some variation from their species-specific consensus sequences. These repetitive motifs are very similar to the sequences flanking human Alu and rodent B1 repetitive elements, although no evidence for the existence of Alu or B1 was found near the peculiar repetitive DNA sequences in apoA-I gene. PMID- 9415808 TI - Effects of the local anesthetic bupivacaine on mitochondrial energy metabolism: change from uncoupling to decoupling depending on the respiration state. AB - The local anesthetic bupivacaine has been found to uncouple oxidative phosphorylation. However, the precise mechanisms of bupivacaine uncoupling have not been elucidated. In the present paper, we demonstrate that the uncoupling effect of the local anesthetic depends on the respiration state. In state 4 respiration (no ADP phosphorylation), bupivacaine acts as a true protonophoretic uncoupler. On the other hand, in state 3-respiration (ADP phosphorylation), bupivacaine induces a change in proton pump stoichiometry and appears to be a decoupler (slip inducer). Moreover, at high concentration, the local anesthetic inhibits the respiratory chain itself. Thus, the divergent reports in the literature on the action of bupivacaine show in fact the various mechanisms by which the local anesthetic may alter mitochondrial energy metabolism. PMID- 9415809 TI - The irradiation of hepatocytes with He-Ne laser causes an increase of cytosolic free calcium concentration and an increase of cell membrane potential, correlated with it, both increases taking place in an oscillatory manner. AB - Isolated hepatocytes were irradiated with Helium-Neon laser (fluence: 0.24 Joules x cm-2, fluence rate: 12 mW x cm-2) and changes of both cytosolic free Ca2+ concentration and cell membrane potential were checked by measuring fura-2 and bis-oxonol fluorescence respectively. Irradiation resulted in an enhancement in cytosolic free Ca2+ concentration that requires the presence of Ca2+ in the phase outside hepatocytes; consistently an increase in cell membrane potential was measured correlated with it. Interestingly, the rate of increase of both cytosolic free Ca2+ concentration and cell membrane potential shows special time dependent features similar to those peculiar of oscillatory processes. PMID- 9415811 TI - Insulin rapidly stimulates ERK2 in the membrane of osteoblast-like UMR-106 cell. AB - We investigated subcellular distribution of ERK2 in osteoblast-like UMR-106 cell and explored to determine if its activities are regulated by insulin. 23%, 34% and 43% of total ERK2 were distributed in membrane, cytosol and nucleus, respectively. Insulin caused 40% increase of ERK2 content in membrane in 10 min whereas it induced approximately 50% decrease of ERK2 in cytosol in 10 min. In terms of kinase activity, insulin stimulated phosphorylation of the membrane associated ERK2 by 2-fold and 1.8-fold in 1 min and 10 min and cytosolic ERK2 by 2.7-fold and 2.3-fold in 1 min and 10 min, respectively. In contrast, the phosphorylation of nuclear ERK2 was stimulated by insulin in time-dependent manner with maximal (3-fold) activity observed at 30 min. Insulin also increased the content of MEK2 in membrane by 2.2- to 2.6-fold in 10 min. MEK2 translocated into membrane in response to insulin may play a role in the activation of the membrane-associated ERK2 via phosphorylation. PMID- 9415810 TI - Inhibition of the proliferative effect of transforming growth factor-alpha by c myc antisense DNA in human ovarian cancer cells. AB - In search of critical genes associated with the mechanism of transforming growth factor-alpha (TGF alpha) action in human ovarian cancer, it was found that TGF alpha stimulates c-myc gene expression in human ovarian NIH:OVCAR-3. The role of c-myc in TGF alpha-stimulated growth of NIH:OVCAR-3 cells was examined by the use of the synthetic antisense-myc phosphorothioate oligonucleotide (OPT). Prior exposure of NIH:OVCAR-3 cells to an antisense-myc OPT inhibited TGF alpha stimulated cell growth and DNA synthesis in a dose-dependent and sequence specific manner over 4 days. c-Myc protein expression was down-regulated in the antisense-myc treated cells. These results demonstrate both the specific and durable effects of the antisense-myc OPT. Furthermore, the results suggest a role for c-myc in TGF alpha-stimulated cell proliferation. PMID- 9415812 TI - Regional heterogeneity of plasma membrane proteins in rat brain. AB - Plasma membranes separated and purified from rat cerebral cortex and cerebellum showed varying protein profiles on SDS-PAGE scanning densitometry. Using highly sensitive two-dimensional gel electrophoresis these proteins were further resolved and an apparent regional heterogeneity of plasma membrane associated proteins was demonstrated. These proteins might be involved in region specific structural and functional aspects in the brain. PMID- 9415813 TI - Role of transglutaminase in keratinization of vaginal epithelial cells in oestrous cycling rats. AB - Aspects of the regulation of calcium dependent transglutaminase (TGase) enzyme in the terminally differentiating vaginal epithelial cells (VEC) from cycling and oestradiol primed ovariectomized (OVX) rats are described. There is a significant increase in the TGase activity and a quantitative rise in its cross-linked sigma(tau-glutamyl)lysine covalent product in the VEC of oestrus rats. A similar phenomena was also evident in the oestradiol primed OVX rats. However, in the VEC of the diestrus/unprimed OVX rats, the enzyme activity and its cross-linked product tend to be at its basal level. An increase of TGase activity in the keratinized layers of the oestrus VEC was observed. Immunohistochemical localization data parallel the biochemical measurements. These findings suggest that TGase activity may be associated with the differential status of the cell and primarily aids in the conferring structural stability and integrity to the stratified VEC. Aggregates of the keratin tonofilament bundles which are covalently cross-linked by the formation of sigma(tau-glutamyl)lysine in the oestrus vagina renders them resistant to denaturant and proteolytic treatments. PMID- 9415815 TI - Sheep testicular and epididymal angiotensin converting enzyme: inhibitions by captopril, lisinopril and enalapril. AB - Inhibition of angiotensin converting enzyme(ACE) in presence of captopril(C), lisinopril(L) and enalapril(E) were investigated in testis and epididymis of sheep using Hip-His-Leu as substrate. Captopril, lisinopril and enalapril were competitive inhibitors of the enzyme from both tissues. Differences in the I50 and Ki values using these three inhibitors reflects the affinities of these inhibitors for the ACE. In addition, the relative potencies of captopril, lisinopril and enalapril were different for testicular ACE(C > L > E) and epididymal ACE(L > C > E). This observation suggests differences between the active sites of the testicular and epididymal ACE which may reflect on their functions in vivo. PMID- 9415814 TI - A new family of plasma membrane polypeptides differentially regulated during plant development. AB - Two cDNAs encoding polypeptides identified in a tobacco leaf plasma membrane fraction prepared by phase partitioning were cloned. The deduced polypeptides, P16 and P17, exhibit a striking primary structure, similar to that of P19, a previously cloned plasma membrane polypeptide. Antibodies raised to the recombinant proteins were used to probe the cellular location of P16, P17 and P19 by means of western blotting of sucrose density-gradient fractions; all three polypeptides were found to be located solely at the plasma membrane. Furthermore, P19 antigen accumulated transiently at the time of floral induction while P16 and P17 antigens accumulated towards the end of the life-cycle. These results together with sequence database searches and multiple-sequence alignments suggest that we have identified a new family of plasma membrane polypeptides that (i) are putatively plant specific and (ii) are differentially regulated during plant development. These polypeptides are termed DREPPs for developmentally regulated plasma membrane polypeptides. PMID- 9415816 TI - Effect of n-3 fatty acids on VLDL production by hepatocytes is mediated through prostaglandins. AB - The mechanism of the hypolipidemic effect of n-3 fatty acids was studied using isolated rat hepatocytes maintained in culture. EPA and DHA caused a significant reduction in the incorporation of 3[H]-leucine into apoB associated with the VLDL produced by hepatocytes in culture when compared to that in presence of palmitic acid. Presence of indomethacin, an inhibitor of cyclo-oxygenase reversed the effect of EPA on VLDL synthesis while diethyl carbamazine an inhibitor of lipoxygenase did not show any effect suggesting that the effect of EPA may be mediated through prostaglandins. This was further tested by invivo experiments where animals were fed fish oil containing diet with and without aspirin, which inhibits formation of prostaglandins. The incorporation of 3[H]-leucine into apo B and 14[C]-acetate into cholesterol of VLDL produced by hepatocytes from aspirin treated animals were significantly high. The reversal of the effect of n-3 fatty acids by agents which inhibit the formation of prostaglandin suggests that the n 3 fatty acids may exert their effect on VLDL production by liver cells through prostaglandins. PMID- 9415817 TI - Antitumor activity of conjugates of the oncofetal protein alpha-fetoprotein and phthalocyanines in vitro. AB - The several conjugates of aluminium and cobalt complexes of phthalocyanines with human alpha-fetoprotein have been synthesized. Their cytotoxic activity against tumor cells and human peripheral blood lymphocytes was studied. The experimental data demonstrate that the cytotoxic activity of alpha-fetoprotein-phthalocyanine conjugates against three types of tumor cells of various origin is much higher (for aluminium and cobalt complexes more than 1000 and 50 times, respectively) in comparison with phthalocyanines themselves. The application of phthalocyanines as conjugates with alpha-fetoprotein makes it possible to markedly enhance the selective toxicity of phthalocyanines against human tumor cells. PMID- 9415818 TI - Inhibition of proteases, myotoxins and phospholipases A2 from Bothrops venoms by the heteromeric protein complex of Didelphis albiventris opossum serum. AB - The antibothropic complex (ABC) from opossum (species Didelphis albiventris) serum was purified by chromatography on DEAE-Sephacel. It showed an acidic character and two polypeptide chains of ca. 45 kDa and 48 kDa, respectively. Lyophilized opossum serum or the ABC (100 micrograms), as well as ethylenediamine tetraacetate (0.25 mumoles) were able to completely neutralise the hemorrhagic effect of 50 micrograms of the desiccated venoms of Bothrops moojeni, Bothrops pirajai and Bothrops jararacussu. The myotoxic (100 micrograms venom in mice) and edematogenic (90 micrograms venom in rats) activities of Bothrops moojeni and Bothrops jararacussu venoms, as well as of the major myonecrotic protein (myotoxin-I) isolated from Bothrops moojeni venom, were also totally inhibited by the ABC (200 micrograms and 270 micrograms, respectively). The lyophilized opossum serum (30 micrograms) and the ABC (30 micrograms) reduced to 50% the phospholipase A2 activity of Bothrops moojeni venom (10 micrograms). The clotting activity of Bothrops alternatus and Bothrops moojeni (20 micrograms) on bovine plasma was also significantly inhibited by the ABC (60 micrograms). PMID- 9415819 TI - Effect of calcium ion channel antagonists on chorionic gonadotropin secretion. AB - By using calcium ion channel antagonists such as Verapamil and Nifedipine, we demonstrated that calcium ion channels are involved in human chorionic gonadotropin (hCG) secretion stimulated by gonadotropin releasing hormone (GnRH). Addition of 1 microM Verapamil (Class II inhibitor) resulted in an inhibition of GnRH-stimulated hCG secretion by placenta without affecting basal release. However, while addition of 10 microM Nifedipine (Class I inhibitor) resulted only in partial inhibition, significant inhibition was observed at 100 microM concentration and above. These results suggest that calcium ion channels in placenta appear to be similar although not identical with the channels reported in the pituitary. PMID- 9415820 TI - Kinetic analysis of the Fenton reaction by ESR-spin trapping. AB - A quantitative analysis of hydroxyl radical (.OH) generated in the Fe(2+) hydrogen peroxide reaction system was explored by a spin-trapping method using 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) combined with electron spin resonance spectroscopy. Based on the numerical analysis of Fenton-related reactions, the reduction of DMPO-OH adduct from 1:1 stoichiometry prominent at high concentrations of Fe2+ was consistent with a reaction model in which a molar amount of hydrogen peroxide was reduced by two molar amounts of Fe2+. Furthermore, time-dependent decrease in DMPO-OH quantity, apparent at much higher concentration of Fe2+, was proved due to the reaction not with Fe2+ but with Fe3+. PMID- 9415821 TI - Protein degradation in red cells exposed to 2,2'-azo-bis(2-amidinopropane) derived radicals. AB - Extensive proteolysis is observed when red blood cells are exposed to free radicals produced in the thermolysis of 2,2'-azo-bis(2-amidinopropane). It is evaluated that nearly one amino terminal group is produced by each free radical introduced into the system. These groups are considered to arise mainly from band 3 fragmentation due to the action of red cell proteinases. Protein fragmentation takes place prior to significant hemolysis or lipid peroxidation, as evaluated by thiobarbituric acid-reactive substances measurements. PMID- 9415822 TI - Fatty acid composition of brown adipose tissue in dietary obese rats. AB - The effects of both dietary obesity and a food deprivation period of 24 hours on fatty acid composition of brown adipose tissue have been investigated. Long time exposure to a hypercaloric high-fat diet such as the cafeteria diet induced an important tissue fatty acid accumulation, mainly for the major saturated and monounsaturated fatty acids. Notable metabolic differences have been observed in the behaviour of control and obese rats facing a food deprivation period: a preferential utilization of the most abundant saturated fatty acids in control rats and a minor response in obese rats, with a greater fat accumulation in the interscapular brown adipose tissue. PMID- 9415823 TI - Evidence for the transfer in culture of [14C]-labelled fatty acids from macrophages to lymphocytes. AB - [14C]-labelled palmitic acid (PA), oleic acid (OA), linoleic (LA) and arachidonic (AA) acids were transferred from macrophages (M phi) to lymphocytes (LY) when equal numbers of the two cell types were co-cultured. The relative degree and amounts of the fatty acids transferred from M phi to LY are as follow: AA (368.57 +/- 21.62) = OA (274.52 +/- 15.41) > LA (42.11 +/- 8.31) = PA (36.53 +/- 2.45). The transfer units are nmol/10(10) M phi/10(10) LY and the values are mean +/- SEM for 7 experiments. The [14C]-radioactivity transferred was mainly directed to the phospholipid fraction of the lymphocytes (85% by PA, 86% by LA, 83% by OA and 79% by AA). In the same order as above, phosphatidylcholine was the phospholipid moiety most heavily labelled (82% by PA, 71% by LA, 66% by OA and 47% by AA). The amount of [14C]-radioactivity transferred to stimulated lymphocytes of thioglycollate treated animals remained unchanged for LA, PA and AA but reduced for OA (71%). The significance of these observations for the immune functions of the cells and resolution of the question of whether some of the [14C]-isotope transfer involves a component of exchange or is unequivocally net fatty acid mass transfer are still being investigated. PMID- 9415824 TI - Cloning of a novel retinoic acid-inducible mRNA from human osteoblast-like cells. AB - Retinoids have long been known to influence skeletal development and bone remodeling. Cells of the osteoblastic lineage play a key role in these processes. In this study we have used the differential display PCR technique to identify retinoic acid (RA)-induced mRNAs in human osteoblast-like cells. We report the cloning and sequencing of one such mRNA, AT-RA 6, which was specifically induced by all-trans RA both in normal human osteoblast-like cells and in MG-63 osteosarcoma cells. Maximal expression was found after 60 min, suggesting that this may be an early response gene. Expression was found in all tissues examined. No homology to known mRNA sequences was detected. PMID- 9415825 TI - IUPAC-IUBMB Joint Commission on Biochemical Nomenclature (JCBN) and Nomenclature Committee of IUBMB (NC-IUBMB). PMID- 9415826 TI - Mechanisms by which insulin and muscle contraction stimulate glucose transport. AB - Insulin binding to its receptor activates a tyrosine kinase that initiates a cascade of signaling events, the initial step being the tyrosine phosphorylation of insulin receptor substrate 1 (IRS-1). Subsequent IRS-1 association and activation of phosphatidylinositiol 3-kinase (PI 3-kinase) is believed to be involved in the events leading to the translocation of glucose transporters (GLUT4) to the plasma membrane resulting in uptake of glucose into the cell. Muscle contractions increase insulin sensitivity, but also stimulate muscle glucose uptake independent of insulin. The contraction signaling pathway is distinct from the insulin pathway because the effect of insulin and contractions on glucose uptake are additive, and contractions do not increase insulin receptor kinase or PI 3-kinase activity. In contrast, studies indicating that contractions cause the translocation of GLUT4 and that both contractions and insulin stimulated glucose transport can be blocked by calcium channel blockers suggest that the two pathways may converge. However, the possibility that two distinct GLUT4 pools may be targeted, one by insulin the other by contractions, indicates that additional research is needed to better define the mechanisms by which glucose transport is stimulated in muscle. PMID- 9415827 TI - Lactate transport and lactate transporters in skeletal muscle. AB - The study of lactate transport in skeletal muscle had until recently been hampered by the lack of suitable sarcolemmal vesicle preparations. Researchers are now at the threshold of developing some very new understandings about the movement of lactate into and out of skeletal muscle with (a) evidence for a lactate transport system in skeletal muscle, (b) the very recent cloning of several monocarboxylate transporter genes, (c) the expression of at least one monocarboxylate transporter protein that facilitates the transport of lactate in heart and skeletal muscle, and (d) the realization that lactate transport can be altered with changes in chronic muscle activity. The MCT1 expression patterns in metabolically heterogeneous skeletal suggests that a primary role of this lactate transporter is to take up lactate into the oxidative muscle fibers where it may be used as a fuel in mitochondrial oxidation. Increments in both MCT1 and lactate transport with training support this role. PMID- 9415828 TI - Anaerobic threshold determination with analysis of salivary amylase. AB - The purpose of this study was to determine the anaerobic threshold from analysis of amylase concentration in total saliva during a laboratory exercise test. Each of 20 healthy young men performed both a submaximal and a maximal test on a treadmill. During the submaximal test, capillary blood and total saliva samples were collected for determination of anaerobic threshold (AT) and saliva threshold (Tsa), respectively. Tsa was defined as the point at which the first continuous increase in amylase concentration occurred during exercise. The results showed no significant difference between values of AT and Tsa when both were expressed either as running velocity or as heart rate. In addition, there existed a high correlation between AT and Tsa (r = .93, p < .001). It was therefore concluded that the analysis of amylase concentration in total saliva during exercise might be used as a valid new method for determining AT. PMID- 9415829 TI - Comparison of a mathematical model to predict 10-km performance from the Conconi test and ventilatory threshold measurements. AB - The purpose of this study was to validate a mathematical model (MM) that evaluates the Conconi test and predicts 10-km race time. In addition, the relationship between ventilatory threshold (Tvent) determined from a laboratory test and heart rate deflection (HRd) from the Conconi test were examined. Seventeen trained runners performed the Conconi test, and performance times were predicted using a MM based on a logistics function. A correlational analysis indicated a highly significant relationship (r = .98, p < .01) between MM predicted time and actual time. Significant relationships were found between velocity at Tvent and HRd (r = .95, p < .01), and predicted times from each method (r = .96, p < .01). Heart rates from Tvent and HRd were also related (r = .79, p < .01). These results suggest that a MM of the Conconi test is a valid method of predicting 10-km performance and is closely related to traditional laboratory measures. PMID- 9415830 TI - The influence of intermittent fatigue exercise on early and late phases of relaxation from maximal voluntary contraction. AB - Twenty-two young male subjects were tested to estimate the behavior of the early and late phases of relaxation from a 3-s maximal voluntary contraction (MVC) under the influence of fatigue. Less demanding and more demanding protocols of intermittent hand grip exercise were used to fatigue muscle. Before and after fatigue, the early and late relaxation time, maximal relaxation rate, and half relaxation time were measured. The results showed that during voluntary movement (a) the early phase of relaxation was independent of the mode of intermittent exercise and did not change significantly after fatigue; (b) the late relaxation time and absolute maximal relaxation rate were slower after both protocols, with the changes more pronounced following the more demanding protocol; and (c) the half-relaxation time and relative maximal relaxation rate were changed only in the more demanding protocol. It is concluded that unlike the relaxation following electrical stimulation of isolated muscle, the early phase of relaxation from voluntary contraction appears to be the most resistant to the type of intermittent fatiguing exercise used in the present study, whereas the late relaxation time was the most sensitive to this type of fatigue. PMID- 9415831 TI - Mechanical properties and behaviour of motor units in the tibialis anterior during voluntary contractions. AB - The present work was carried out to analyse the properties and behaviour of Tibialis anterior motor units (MUs) during voluntary contractions in humans. A total of 528 single MU mechanical properties was recorded in 10 subjects by means of the spike-triggered averaging (STA) technique. MU recruitment thresholds and discharge frequencies were recorded during linearly increasing maximal voluntary contraction (MVC). The results indicate a mean (+/- SD) MU torque of 25.5 +/- 21.5 mN.m. and a mean time-to-peak of 45.6 +/- 13.6 ms. A comparison of the average MU twitch torque with that of the muscle allowed an estimate of about 300 MUs in the Tibialis anterior. A positive linear relationship was recorded between the MU twitch torque and the recruitment threshold. The mean minimal and maximal discharge frequencies of MUs were 8.4 +/- 3.0 Hz and 33.2 +/- 14.7 Hz, respectively. The results of the present work indicate that MU behaviour during voluntary contractions is different in the tibialis anterior and in the adductor pollicis. PMID- 9415832 TI - A new formula for population-based estimation of whole body muscle mass in males. AB - A new equation to estimate muscle mass in males was developed using parameters common to the 1981 Canada Fitness Survey and the male cadaver data of Martin et al. (1990b). The cadavers (N = 12) were randomly divided into two groups. The equation was developed on cadaver Group A and then validated on Group B. Once the equation with the most suitable variables was validated on Group B, it was redeveloped on combined data from Groups A and B. The final equation is as follows: muscle mass (gm) = Ht (0.031MUThG2 + 0.064CCG2 + 0.089CAG2) - 3,006; adjusted R2 = .96, SEE = 1,488 gm, F = 87.5, p = .0001. Variables (in cm) were Ht, height; MUThG, modified upper thigh girth; CCG, corrected calf girth; and CAG, corrected arm girth. The predictive ability of this equation was comparable to the original equation of Martin et al. (1990b) and can be a valuable tool for muscle mass estimation of male subjects in the 1981 Canada Fitness Survey. PMID- 9415833 TI - Neonatal hypoglycemia, Part I: Background and definition. AB - Hypoglycemia in the neonate remains a common problem. The association of low blood glucose concentrations and abnormal development has prompted extensive research into the anticipation, evaluation, and treatment of neonatal hypoglycemia. Glucose homeostasis in the fetus and neonate is a developmentally regulated dynamic process involving a number of intricate physiologic mechanisms. In addition, the determination of glucose concentrations is dependent upon both the type of tissue analyzed and the limitations of the specific method employed. The complexity of glucose metabolism makes it difficult to precisely define "normal" and "abnormal" glucose levels in preterm and term neonates. PMID- 9415834 TI - Paradoxical reactions in children associated with midazolam use during endoscopy. AB - All 2,617 children who received midazolam and meperidine for a variety of endoscopic procedures were monitored for the development of adverse behavioral problems. Thirty-six (1.4%) of the children (ages 1-17 years) experienced a paradoxical behavioral reaction, which consisted of inconsolable crying, combativeness, disorientation, dysphoria, tachycardia, agitation, and restlessness. The reaction occurred at a mean of 17 minutes after the administration of midazolam. Following treatment with flumazenil, the reaction dissipated within a mean of 14 minutes. Three of the 36 patients underwent additional endoscopic procedures utilizing only meperidine. No similar reaction was observed in these patients. Awareness of the reaction and prompt administration of flumazenil decreased the duration of the reaction. PMID- 9415835 TI - An analysis of group versus individual child health supervision. AB - This study compares the effectiveness of group health supervision (three or four families counseled simultaneously) with traditional visits in conveying knowledge of child health and development, increasing perceived maternal support, and mitigating maternal depression. Subjects were recruited from a predominantly white, middle-class, suburban/rural pediatric practice. Twenty-five families were allocated to group health supervision and 25 to individual visits. A questionnaire covering knowledge of child health and development (CHDQ), the Maternal Social Support Index (MSSI), and the Center for Epidemiologic Studies Depression Scale (CESD) were administered to both groups before their 2-month and after their 10-month visits. A subset of these charts was reviewed for problem visits between 2 and 6 months. As compared with families having traditional visits, families who received the group intervention did at least as well in acquiring knowledge of child care and development and, although not statistically significant, tended to recover from postpartum depression faster and deal better with minor illnesses. The investigators found group child health supervision to be a pleasant and effective method of health care delivery. PMID- 9415836 TI - Resident documentation of diagnostic impression in sexual abuse evaluations. AB - In sexual abuse evaluations, the documentation of the examiner's diagnostic impression is essential. If the diagnostic impression is not documented, the examiner will have to rely on memory rather than the medical record when called to testify. The purpose of this study was to determine whether pediatric residents adequately document their diagnostic impression in child sexual abuse evaluations. We performed a three-year retrospective chart review from patients 0 17 years of age who were evaluated at our emergency room for suspected sexual abuse. We reviewed 1,487 charts for historical information, physical findings, and diagnostic impression. Physical findings were categorized as normal, nonspecific, suggestive, or indicative of penetration. In 77% of cases (N = 256) with hymenal findings indicative of penetration and 84% of cases (N = 31) with vaginal findings indicative of penetration, residents recorded no impression or a nonspecific impression. Results were similar for vulvar and rectal findings indicative of penetration. Residents fail to document an adequate interpretation of their physical examinations in sexual abuse evaluations. PMID- 9415837 TI - Mold allergy is a risk factor for persistent cold-like symptoms in children. AB - In winter, children with mold allergy may develop persistent cold-like symptoms (PCLS) that often defy conventional therapy. To investigate the cause of PCLS, we enrolled 44 children (25 with PCLS and 19 controls) in a 2-year study to compare their clinical symptoms and the mold count in their homes. Children with PCLS had a higher percent of eosinophils in nasal smears as compared with those without PCLS (32% vs 26%). On a scale of 0 to 3, the PCLS group had higher symptom scores (P < 0.001 for all symptoms): bloodshot eyes (2.92 vs 0.79), mouth breathing (2.04 vs 0.68), rhinorrhea (2.48 vs 0.89), nasal voice (2.68 vs 1.00), postnasal drip (2.64 vs 0.47), and headache (2.72 vs 0.53) than the non-PCLS group. The clinical scores also correlated significantly with the mold count in the home (the r value ranged from 0.6716 to 0.7450). We conclude that management of children with PCLS should include decreasing humidity and enforcing environmental control to eradicate mold from inside the homes. PMID- 9415838 TI - Foreign body in the tracheobronchial tree. AB - A 20-year experience with the treatment of 74 patients (83.8% children) for foreign body aspiration is reviewed. The object of this review is to show the clinical manifestations, the radiological findings, the nature and distribution in the bronchial tree, and complications due to longstanding (months or years) foreign bodies in the bronchial tree. The most common foreign bodies found were peanuts (13.5%), corn (13.5%), and beans (13.5%). The most frequent clinical manifestation was choking (67.5%), and the most frequent radiological finding was atelectasis (41.8%). The most serious complication was bronchiectasis needing resection in six patients who had the foreign body retained for years in the bronchial tree. In conclusion, in spite of an obvious foreign body in the tracheobronchial tree many cases are not diagnosed, and a longstanding foreign body in the airway may be responsible for irreversible complications. PMID- 9415839 TI - Adverse reactions to cromolyn sodium: patient report and review of the literature. PMID- 9415840 TI - Pseudotumor cerebri: a presenting manifestation of Addison's disease. PMID- 9415842 TI - Congenital adrenal hyperplasia and bilateral ovarian cysts in a neonate. PMID- 9415841 TI - Lemierre's syndrome: new insights into an old disease. PMID- 9415843 TI - Mild thalassemia intermedia resulting from a new insertion/frameshift mutation in the beta-globin gene. AB - Hematological investigation of an antenatal patient led to the identification of a new beta-thalassemia mutation involving the net insertion of eight nucleotides into exon 2 of the beta-globin gene. As a result of the shift in the protein reading frame, this gene codes for an elongated beta-globin chain (159 amino acids) with an abnormal amino acid sequence beyond residue beta 99. There is no evidence of any abnormal hemoglobin in the circulation. The patient has a mild form of beta-thalassemia intermedia with moderate anemia, evidence of iron overload, severe red cell morphological changes, a significant reticulocytosis, and a marked increase in the proportion of fetal hemoglobin. PMID- 9415844 TI - Hb Siirt [beta 27(B9)Ala-->Gly]: a new, electrophoretically silent, hemoglobin variant. PMID- 9415845 TI - A novel beta-thalassemia mutation [codon 45(-T)] in a Pakistani family. PMID- 9415846 TI - The acute effect of stenting with the nitinol self-expanding coil stent: preliminary experience. AB - BACKGROUND: The acute angiographic results with the self-expanding nitinol stent have not been reported. We aim to provide angiographic data of the effect of self expansion and balloon assistance on the results. This is analyzed with respect to stent gain, arterial- and stent-recoil. METHODS AND RESULTS: The self-expanding nitinol coil stent is inherently different than balloon-expandable stents in its mechanism of deployment and the way that radial arterial expansion is achieved. Between January 1995 and June 1996, 86 stents were deployed in 64 patients undergoing elective angioplasty at the Rambam Medical Center, Haifa, Israel. The stent deployment procedure involved stent release assisted by high pressure balloon dilatation. The baseline, post-balloon, post-stenting and post-stent dilatation characteristics were recorded with similar views, digitized to a PC and analyzed by image processing software. Using computerized analysis, arterial- and stent-recoil and stent gain were calculated for the average stented segment lesion (0.48 +/- 0.42, 0.22 +/- 0.37, 0.28 +/- 0.37, respectively). Balloon angioplasty increased the minimal luminal diameter from 1.07 +/- 0.73 mm at baseline, to 2.24 +/- 0.57 mm; stent deployment further increased the diameter to 2.63 +/- 0.48 mm, and within-stent balloon dilatation to 2.96 +/- 0.62 mm. CONCLUSIONS: The self-expanding nitinol stent exerts its effect on both the MLD and the average stented diameter through its intrinsic radial force aided by post deployment within-stent balloon dilatation. A significant correlation was found between stent gain and arterial recoil (slope = 0.59, r = 0.68, p < 0.001) but not with stent-artery recoil. Therefore, with the negligible effect of stent recoil, the acute benefit of the nitinol stent is directly proportional to arterial recoil, a feature which is also common to balloon-expandable stents. PMID- 9415847 TI - Towards a geometrically correct 3-D reconstruction of tortuous coronary arteries based on biplane angiography and intravascular ultrasound. AB - At present, 3-D reconstructions of coronary vessels are generated from intravascular ultrasound (IVUS) by stacking up ECG-gated segmented IVUS frames of a pullback sequence. This simplified approach always results in straight vessel reconstructions and, therefore, gives an incorrect representation of tortuous coronary arteries. A more realistic reconstruction of tortuous vessels may be obtained by data fusion with biplane angiography. The 3-D course of the vessel is first derived from the angiograms and then combined with the segmented IVUS images. In this paper, we focus on two problems associated with the data fusion method: The definition of the pullback path and the estimation of the IVUS catheter twist during pullback. A robust algorithm for calculation of tortuosity induced catheter twist is reported that is based on sequential triangulation of the 3-D pullback path. The method is analyzed with computer simulations and validated in helical vessel phantoms. A largely automated data fusion approach is proposed and applied to tortuous coronary arteries in cadaveric pig hearts. PMID- 9415848 TI - Dynamic on-line quantification of biventricular function with acoustic quantification (AQ). Validation, reproducibility and normal values of a new echocardiographic approach. AB - OBJECTIVES: Acoustic quantification (AQ), a recently developed ultrasonic integrated backscatter imaging system providing on-line measurements of ventricular cavity areas and their functional indexes, was validated in comparison to angiography and Doppler derived systolic dP/dt. Normal AQ-reference values were established. METHODS AND RESULTS: 1. In 45 patients undergoing heart catheterization, AQ derived areas in end-diastole (EDA), end-systole (ESA) and the resulting fractional area change (FAC) in apical 2- and 4-chamber view were compared to the corresponding biplane angiographic data. All correlations yielded significant values (p < 0.0001; EDA: r = 0.90, SEE = 2.6 cm2; ESA: r = 0.91, SEE = 2.2 cm2; FAC: r = 0.90, SEE = 4.1%). However, AQ-areas were underestimated by about 25%. 2. In 36 patients with mitral regurgitation AQ-FAC and AQ derived systolic dA/dt were compared to the Doppler derived systolic dP/dt, yielding significant correlations with r = 0.91 and r = 0.87; p < 0.0001. 3. In 50 healthy subjects, AQ derived EDA, ESA and FAC averaged 25.7 +/- 4.9, 14.7 +/- 3.3 cm2 and 43.2 +/- 4.8% for the left, and 17.1 +/- 3.8, 9.0 +/- 2.9 cm2 and 47.3 +/- 9.2% for the right ventricle. For EDA normalized peak filling (PFR) and ejection rates (PER) yielded 2.7 +/- 0.28 and -2.4 +/- 0.42 EDA/sec for the left and 3.4 +/- 0.74 and -2.9 +/- 0.62 EDA/sec for the right ventricle. The interobserver and day to-day variability of AQ in healthy subjects and cardiac patients was low for EDA, ESA and FAC (< 12%) and higher for PFR and PER (< 20%). CONCLUSION: In comparison to angiography AQ reliably quantitates on-line left ventricular fractional area change, although AQ-areas are underestimated. AQ offers reproducible values of systolic and diastolic function and a new approach to cardiac patients. PMID- 9415849 TI - Morphology of chronic coronary occlusions and response to interventional therapy- a study by intracoronary ultrasound. AB - OBJECTIVES: Balloon angioplasty of chronic coronary occlusions has a low procedural success and a high recurrence rate. Better tomographic insights into the lesion morphology may improve the interventional strategy and results. METHODS: Intracoronary ultrasound was used during the recanalizaton procedure of 45 chronic coronary occlusions (2 weeks to 14 months; average 3.4 months) to determine the lesion morphology and to assess the angioplasty result. The luminal area and the plaque burden were measured proximal and distal to the occlusion, and within the occlusion. The ultrasonographic characteristics of the occlusive lesions were compared to 45 nonocclusive lesions of age-matched patients with stable angina pectoris. RESULTS: Occlusive lesions were more often echodense as compared to nonocclusive lesions (35% vs. 20%; p = 0.10). In chronic occlusions a multi-layered plaque morphology was observed in 22%, and this morphology was not found in nonocclusive lesions. Angiographic characteristics were not related to the ultrasonographic morphology of the lesion. Despite similar vessel areas in occlusive and nonocclusive lesions, the balloon size selected according to the angiographic image was underestimated in occlusive lesions. Based on the quantitative ultrasound measurement the balloon size was increased from 2.6 +/- 0.3 mm to 3.3 +/- 0.5 mm in 53% of the lesions. This resulted in an increase of the luminal area from 3.51 +/- 0.92 to 5.08 +/- 1.43 mm2 (p < 0.001). The acute recoil after balloon angioplasty was similar (34 +/- 18%) in hypodense and echodense plaques, but was significantly higher in lesions with a multi-layered plaque morphology (49 +/- 22%; p < 0.05). In 19 patients with severe dissections or extreme acute recoil (residual stenosis > 50%) the use of a stent increased the luminal area from 3.94 +/- 0.81 to 7.51 +/- 1.71 mm2 (p < 0.001). CONCLUSIONS: Intracoronary ultrasound demonstrated a multi-layered plaque morphology in one fourth of the chronic occlusions. This type of plaque was associated with a significant acute recoil. The presence of diffuse atherosclerosis in neighbouring segments of chronic coronary occlusions leads to underestimation of the balloon size. Quantitative assessment by intracoronary ultrasound helped to optimize the balloon size leading to a significant luminal area gain. The detection of excessive acute recoil should be considered an indication for stent deployment. PMID- 9415850 TI - Dynamic stress echocardiography for evaluating anti-ischemic drug profiles in post-MI patients. AB - Exercise ECG is an established method of evaluating the anti-ischemic properties of drugs. However, there are considerable methodologic limitations to this procedure and its use is restricted to patients with exercise-provoked ECG alterations which can be interpreted as ischemia. The principal, earlier onset of wall motion abnormalities according to the ischemic cascade can be detected by stress echocardiography and might be utilized as a pharmacological stress testing modality. Sixteen consecutive patients (15 men, one woman; 53 +/- 9 years old) with angiographically proven coronary artery disease (8 with one-, 5 with two-, and 3 with three-vessel disease) and exercise-induced wall motion abnormalities were examined by dynamic stress echocardiography (50 watt followed by 20-watt increases/min). Anti-ischemic drugs were withdrawn prior to and on day 1; on the following day 2, 0.2 microgram/kg/min nisoldipine was infused intravenously during the test after a 3 micrograms/kg bolus was given. At maximum comparable workload 15/16 patients showed an improved wall motion score on treatment (day 1: 22.9 +/- 4.9 vs day 2: 20.0 +/- 3.9; normal score: 12; one-sided binomial test: p = 0.0003). Eight of 16 patients demonstrated ST-segment deviations on day 1 and day 2. The double product did not differ at any workload stage until the maximum of 130 watt (day 1: 14,101 +/- 3140 vs day 2: 13,365 +/- 2865; n.s.). Dynamic stress echocardiography seems to be a valuable tool in pharmacologic stress testing and in terms of accuracy is supposed to be superior to conventional exercise ECG. Nisoldipine reduces exercise-induced wall motion abnormalities in patients with and without exercise-induced ECG alterations. The data result from a controlled pilot study, and further studies are required to confirm these promising methodological and therapeutic findings. PMID- 9415851 TI - Factors influencing the diagnostic accuracy of dobutamine stress echocardiography. AB - BACKGROUND: While Dobutamine stress echocardiography is a well established tool, the range of the diagnostic accuracy found in the literature is rather large. The main reason for this is the fact, that different test protocols were used. Aim of this study was to assess the effects of both addition of atropine as well as consideration of a hyperdynamic response while interpreting the stress echocardiogram on the diagnostic accuracy. METHODS AND RESULTS: 120 consecutive patients were examined and divided into the following groups: A) achieving their age predicted heart rate with dobutamine, B) termination of the test due to ischemia, C1) negative test without reaching the predicted heart rate, and C2) C1 following addition of atropine. All of the echocardiograms were analyzed twice: 1) regarding the lack of a hyperdynamic response to dobutamine as ischemia (Hyper analysis), and 2) ignoring the hypercontractility (Conventional analysis). The accuracy of A and B were 88% and 90% resp. Group C1 had a very poor accuracy of 60%. This rose significantly (p < 0.01) after atropine (C2 = 84%), without leading to an increase of adverse effects. Conventional wallmotion analysis lead to an overall accuracy of 87% (groups A, B, and C2), while Hyper analysis showed an accuracy of 90% (p < 0.01). CONCLUSIONS: To achieve a high accuracy Dobutamine stress echocardiography should always be combined with atropine to reach a target heart rate. The wallmotion analysis should be based on the assumption that a hyperdynamic response to dobutamine is normal, while its lack is indicative of ischemia. PMID- 9415852 TI - Localization and determination of infarct size by Gd-Mesoporphyrin enhanced MRI in dogs. AB - BACKGROUND: Accurate localization and sizing of a myocardial infarction are necessary for clinical decision making and even more in research. Gd Mesoporphyrin enhanced magnetic resonance imaging (MRI) was recently shown to specifically delineate necrosis in liver tumors, renal and muscle necrosis and myocardial infarction in rats. In this study, we investigated this technique's potential to accurately delineate myocardial infarction in a larger animal species, the dog. METHODS: Myocardial infarction was induced in 8 dogs by ligation of the left anterior descending coronary artery, 4 of which were reperfused after 3 hr Gd-Mesoporphyrin (0.05 mmol/kg) was injected intravenously 210 min after the onset of ischemia (n = 6) or after 24 hr in 2 dogs with non reperfused infarctions. MRI was performed 10 hr after administration of Gd Mesoporphyrin. In vivo MRI consisted of EKG-triggered, respiratory gated T1 weighted spin echo and segmented turboFLASH long and short axis measurements. Post-mortem, a spin echo short axis measurement was repeated. Infarct size was determined planimetrically by TTC staining of left ventricular slices. RESULTS: In all instances, there was a very close qualitative agreement between the MRI and TTC defined myocardial infarction. Quantitatively, the linear regression from post-mortem MRI to TTC determined infarct size yielded a result very close to the line of identity (regression coefficient: 0.980 +/- 0.026, p < 0.000001, adjusted R2 = 0.964). CONCLUSION: We conclude that Gd-Mesoporphyrin enhanced MRI is a promising tool for the accurate delineation of myocardial infarction. PMID- 9415853 TI - Social determinants of experienced anger. AB - We investigated two social determinants (i.e., availability of social support and status differentials of the provocateur) for the degree of perceived anger in two populations. Because no suitable tool was available, the conceptual and psychometric development and validation of a new vignette-based measure for anger level (STandardized Experience of Anger Measure, STEAM) is described first. Two versions of STEAM were developed: one for students and one for community-living adults. Through a series of four studies, two sets of a 12-item vignette-based questionnaire were developed and validated. The resulting test had excellent test retest stability and high internal consistency. Using the new STEAM measure, a variety of analyses were conducted to test the hypothesized influence of social determinants of anger. In the student sample, presence of social support was associated with lessened anger, and in both samples decreasing status of the provocateur also led to lessened anger arousal. In addition, findings in both samples revealed that social support reduced anger when the provocateur was of higher status relative to situations of equal and lesser status. In the community sample, the availability of support was associated with greater intensity of the anger experience in the lesser status condition than in the equal or greater status condition. No gender main effects or interactions were noted. PMID- 9415854 TI - A replicated prospective investigation of life stress, coping, and depressive symptoms in multiple sclerosis. AB - Life stress and coping responses jointly contribute to psychological adjustment in many chronic illness populations, but their significance in multiple sclerosis (MS) has not been extensively investigated. Physical disability, cognitive status, negative life stress, coping strategies, and depressive symptoms were prospectively assessed in 27 adults with definite or probable MS. Of the original subjects, 22 provided two additional assessments at 6-month intervals. After accounting for cognitive status and physical disability, life stress was positively correlated with current as well as future depressive symptoms; the prospective relationship was replicated within the second pair of prospective data waves. Escape avoidance was the only coping strategy that added to the prediction of future mood symptoms, but this was not replicated. Results suggest that MS-related depressive symptoms are a function of prior disease-related impairment, life stress, and possibly escape avoidance coping. PMID- 9415855 TI - Four-year stability of cardiovascular reactivity to psychological stress. AB - The 4-year stability of cardiovascular responses to laboratory psychological stress (mental arithmetic) was examined in 75 adults. The stability coefficients were .76 for heart rate (HR) change and .81 for absolute HR, .66 for systolic blood pressure (SBP) change and .52 for absolute SBP, .16 for diastolic blood pressure (DBP) change and .27 for absolute DBP. Males had greater SBP and DBP reactivity than females in the first session, but this reactivity decreased by the 4-year follow-up session (which was not the case for women). PMID- 9415856 TI - Defensiveness and perception of external inspiratory resistive loads in asthma. AB - This study examined the relationship between defensiveness, as measured by the Marlowe-Crowne Social Desirability Scale (MCSDS), and the perception of an externally applied respiratory resistance among people with asthma. Thirty asthmatic adults breathed through nine levels of inspiratory resistive load. Participants higher on the MCSDS were less accurate than others in psychophysical magnitude estimates of resistive load, and showed a reduced relationship between the physical load and the quality of respiratory sensations associated with exposure to the resistors. Defensive subjects also showed a differentially high increase in correlation between unpleasantness of respiratory sensations and resistance levels after receiving parenteral naloxone. These findings are consistent with the hypothesis that defensiveness may increase risk of asthma morbidity, due to inaccuracy in detecting sensations of dyspnea during asthma exacerbations. The inaccuracy may be caused by elevated endogenous opioids among defensive individuals. PMID- 9415857 TI - Hypertension and sickness absence: the role of perceived symptoms. AB - The association between perceived symptoms and absenteeism was examined in five groups of employed adults: normotensives, unaware hypertensives, aware and untreated hypertensives, aware and treated hypertensives, and falsely aware normotensives. Aware hypertensives (untreated and treated) and falsely aware normotensives had a higher average of perceived symptoms than normotensives, whereas unaware hypertensives had lower. The absenteeism rate across the groups showed a similar pattern. A significant interaction of perceived symptoms by study group on absenteeism was uncovered. Hypertensives and falsely aware normotensives who reported a low level of symptoms were not absent more than their normotensive counterparts. However, aware hypertensives and falsely aware normotensives who perceived a high symptoms level showed higher absenteeism than unaware hypertensives and normotensives with a similar level. This suggests that aware hypertensives have a greater tendency than both normotensives and unaware hypertensives to equate their symptoms with ill health and to act accordingly. Special attention should be directed to aware hypertensives who perceive a threat to their health. PMID- 9415858 TI - Determinants of psychological distress and its course in the first year after diagnosis in rheumatoid arthritis patients. AB - In order to examine determinants of psychological distress and its course in the first year after diagnosis in rheumatoid arthritis patients, self-report data and clinical and laboratory measures were collected in 91 patients (70% female, mean age 57 years) shortly after diagnosis and 1 year later. Multiple regression analysis indicated that sex, pain and functional status, disease impact on daily life, life events, and perceived social support were related to psychological distress (anxiety and depressed mood) shortly after diagnosis. Coping strategies were related to distress levels only 1 year later. Multiple regression analysis of change in anxiety and depressed mood revealed that a decrease of psychological distress after 1 year could be predicted by male sex, an initially less severe inflammatory activity and an initially more extended social network. In addition, a decrease in distress was related to parallel improvements in clinical status. Results indicate the importance of a multimodal assessment of demographic variables, clinical and life stressors and social resources for the understanding of distress and the identification of risk factors in the first stage of the disease. Personal coping resources appear to become more important predictors of distress in a later phase of the disease. PMID- 9415859 TI - Transolecranon fracture-dislocation of the elbow. AB - OBJECTIVE: To characterize the prevalence, morphology, and prognosis of anterior (transolecranon) fracture-dislocations of the elbow. DESIGN: Retrospective case series. SETTING: A consecutive series of thirteen patients from a single level one trauma center, plus four patients from the practices of two of the senior authors. PATIENTS: Three of seventeen patients had simple, oblique fractures of the olecranon, and fourteen had complex, comminuted fractures of the proximal ulna, including fragmentation of the olecranon in seven patients, large coronoid fragments in eight patients, and segmental fractures of the ulna in six patients. Fourteen patients were male and three were female, with an average age of thirty eight years (range, 18 to 78 years). INTERVENTION: All fractures were treated by open reduction and internal fixation. Two one-third tubular plates had to be revised to 3.5-millimeter dynamic compression plates within six weeks of the initial operation. MAIN OUTCOME MEASURE: Elbow performance rating of Broberg and Morrey. RESULTS: At an average follow-up of twenty-five months, overall outcome was rated as excellent in seven patients, good in eight, and fair in two. Mild posttraumatic arthritis was noted in only two patients. Large coronoid fragments and extensive comminution of the trochlear notch did not preclude a good result provided that stable, anatomic fixation was achieved. CONCLUSIONS: Anterior elbow dislocations occur most often as a fracture-dislocation in which the distal humerus is driven through the olecranon, thereby causing a complex, comminuted fracture of the proximal ulna. This injury is frequently confused with anterior Monteggia lesions by virtue of the readily apparent radiocapitellar dislocation. Stable restoration of the appropriate contour and dimensions of the trochlear notch of the ulna will lead to a good result in most cases. PMID- 9415860 TI - Regeneration of diaphyseal bone defects using resorbable poly(L/DL-lactide) and poly(D-lactide) membranes in the Yucatan pig model. AB - OBJECTIVES: To determine the effects of tubular resorbable polymer membranes on the healing of a segmental diaphyseal bone defect. DESIGN: A randomized prospective study using the minipig model. Animals were evaluated with in vivo roentgenograms on a biweekly basis until explanted at twelve weeks. SETTING: After surgery, animals were allowed unrestricted activity and weight bearing between twenty-four and forty-eight hours. ANIMALS: Fifteen yearling Yucatan minipigs. INTERVENTION: A 2.5- to 3.0-centimeter mid-diaphyseal defect was created in the middle third of the radius. Animals were assigned in groups of three to receive the following implants: (a) poly(L/DL-lactide), (b) poly(L/DL lactide)-CaCO3, (c) poly(D-lactide), (d) poly(D-lactide)-CaCO3, and (e) an untreated defect. No adjunctive internal or external fixation was used as the ulna was left intact. MAIN OUTCOME MEASURES: The limbs were studied with in vivo anterior-posterior and lateral radiographs at biweekly intervals for the presence and pattern of bone formation. All limbs were explanted at twelve weeks postimplantation for methyl-methacrylate embedding and histologic and microradiographic study. RESULTS: The bone defects covered with membranes were completely reconstituted by six to eight weeks. Untreated defects healed with less bone formation and in a more disorganized pattern. Histologic evaluation of the implants demonstrated that the entire lumen of the implant was filled with bone, with some periosteal bone formation occurring on the outer surface of the membrane. There was direct apposition of bone onto the membrane surface or minimal fibrous tissue interposition between membrane and new bone. There was no foreign body or adverse reaction to the membrane. Untreated defects showed woven bone formation with clefts and irregularly shaped margins occupied by fibrous tissues or surrounding muscle tissues. CONCLUSIONS: This study supports the concept that a membrane enhances bone defect healing by excluding nonosseous tissues from a defect and providing structural scaffolding for periosteal and endosteal bone regeneration. PMID- 9415861 TI - MR imaging of biodegradable polylevolactide osteosynthesis devices in the ankle. AB - OBJECTIVE: To assess the feasibility of magnetic resonance (MR) imaging in the postoperative follow-up after internal fracture fixation using biodegradable polylevolactide (PLLA) plugs and to investigate the MR characteristics of these devices. STUDY DESIGN AND METHODS: MR findings in ten patients with displaced malleolar fractures treated by internal fixation using absorbable PLLA plugs were evaluated after three different postoperative periods. The average postoperative follow-up time was thirty months for four patients, forty-two months for another four patients, and fifty-one months for the remaining two patients. RESULTS: On T1-weighted coronal images, the geometry of the PLLA plug was clearly visible in all cases, without signs of fatigue failure or absorption. The host-to-tissue area between the deployed two fins of the plug showed higher signal intensity than the surrounding cancellous bone on fat-saturated proton density (PD) and turbo inversion recovery (tIR) images. This area had signal intensity similar to articular cartilage on T1-weighted coronal images. In none of the cases could any fluid accumulation be seen around the plug. In all ten cases, a thin rim with signal intensity similar to the area between the deployed fins was detected around the PLLA plug on fat-saturated axial PD images. On fat-saturated T2 and tIR sequences, this rim was less clearly detectable in all cases. No differences in the signal intensity or geometry of the PLLA plug on the MR images emerged between the three patient groups with mean follow-ups of thirty, forty-two, and fifty-one months. No artifacts produced by the implants were seen on any of the MR images. A biopsy specimen obtained at a reoperation necessary seventeen months postoperatively showed no signs of degradation of the PLLA plug. CONCLUSIONS: MR imaging can visualize PLLA implants within bone. It also shows, without artifacts, the tissue interaction between the artificial biodegradable material and bone tissue in humans. PMID- 9415862 TI - Biomechanical comparison of fixation methods in transverse olecranon fractures: a cadaveric study. AB - OBJECTIVES/HYPOTHESIS: Our null hypothesis was that no difference in fracture displacement would be detected between traditional monofilament wire and Kirschner wire placement versus three modified tension-band techniques for transverse olecranon fractures. STUDY DESIGN: A nested form of the repeated measures design with twenty-two paired embalmed elbows (subjects grouped by sex and nested within the fracture method). METHODS: Transverse osteotomies were created at the olecranon and stabilized with four techniques. One hundred cycles of loading were applied to achieve a peak flexion bending moment at the fracture of nine newton-meters. At the onset of testing, the triceps tendon was anchored at an initial elbow flexion angle of 70 degrees. RESULTS: When using a monofilament figure-eight loop, oblique Kirschner wire placement into the anterior ulnar cortex provided greater resistance to tensile force than intramedullary Kirschner wires (p = 0.04). With intramedullary Kirschner wire placement, 1.6-millimeter-diameter braided cable in both figure-eight (p < 0.0001) and circular loop (p < 0.0001) designs allowed less fracture displacement than did the 1.0-millimeter-diameter monofilament wire. There was no difference between figure-eight and circular loop configurations when using braided cable (p = 0.98). CONCLUSIONS: In transverse noncomminuted olecranon fractures, fixation with monofilament wire is superior with Kirschner wire placement into the anterior ulnar cortex. With intramedullary Kirschner wires, fixation using braided cable is significantly improved over that with monofilament wire. When using braided cable, figure-eight and circular loop designs allow similar displacements. Braided cable or anterior cortical Kirschner wire purchase increases the stability of fixation over that achieved with the traditional method. PMID- 9415863 TI - Open reduction internal fixation of displaced transverse patella fractures with figure-eight wiring through parallel cannulated compression screws. AB - OBJECTIVE: To evaluate the clinical results of transverse patella fracture fixation with a tensioned anterior figure-eight wire placed through parallel cannulated screws. DESIGN: Prospective, clinical. PATIENTS: Ten patients with displaced transverse patella fractures were managed with a standardized rehabilitation protocol that employed early continuous passive knee motion. OUTCOME MEASURES: Time to clinical and radiographic union, Hospital for Special Surgery Knee Scores, and comparisons with literature cohort studies. RESULTS: Clinical union occurred at an of average eight weeks and radiographic union at a mean of thirteen weeks postoperatively. Early continuous passive motion over a restricted are did not compromise the quality of fracture reduction, even though the majority of patients were elderly and had osteopenic bone. Subjective and functional results using Hospital for Special Surgery Knee Scores were comparable to previously published reports, with 70 percent achieving an excellent or good outcome. CONCLUSIONS: The described fixation technique for transverse patella fractures had clinical results equivalent to reports of patella fractures fixed with modified tension band wiring. Advantages included a low-profile construct that caused lesser degrees of implant irritation to local soft tissue structures, was compatible with the use of early restricted motion, and afforded a method to salvage three cases in which traditional tension band wiring failed to maintain an anatomic reduction in oteoporotic bone. PMID- 9415864 TI - Hip screw augmentation with an in situ-setting calcium phosphate cement: an in vitro biomechanical analysis. AB - OBJECTIVES/HYPOTHESIS: This study was performed to determine whether a new, in situ-setting calcium phosphate cement would have sufficient mechanical integrity to reinforce compression screw fixation of unstable intertrochanteric fractures. We compared the cut-out resistance of screws augmented with calcium phosphate cement to the cut-out resistance of screws augmented with polymethylmethacrylate (PMMA). We used PMMA as the standard for comparison because it is currently used clinically. Our hypothesis was that initial fixation strength with PMMA and calcium phosphate cement augmentation would not be significantly different from one another. STUDY DESIGN: Cut-out testing of compression hip screws in paired human cadaveric proximal femurs was performed before and after augmentation with PMMA or calcium phosphate cement. Bilateral testing was performed to allow pairwise comparisons of the materials used for augmentation, and repeated testing was done to provide an internal control for the effects of bone quality. The initial fixation of screws augmented with calcium phosphate cement was compared with that of screws augmented with PMMA. METHODS: Ten paired human femurs (mean age, 75 +/- 9.2 years) were implanted with Richards AMBI compression hip screws. Basicervical osteotomies were then performed, yielding isolated proximal fragments for mechanical testing. Preaugmentation cut-out tests were performed under displacement control, with cut-out continuing to five millimeters at two millimeters per second. The screws were then removed, and the screw tracks were filled with 2.0 cubic centimeters of PMMA (one side) or calcium phosphate cement (contralateral side). After augmentation, the screws were reinserted and the cements were allowed to harden for twenty-four hours. Postaugmentation testing followed the protocols for preaugmentation testing, and the initial fixation strength of screws augmented with calcium phosphate cement was compared with the initial fixation strength of screws augmented with PMMA using a two-way repeated measures analysis of variance. RESULTS: The cut-out behavior of screws augmented with calcium phosphate cement was not significantly different from the cut-out behavior of screws augmented with PMMA. With calcium phosphate cement, yield strength increased by 15.8 percent (from 1,354 +/- 632 newtons to 1,568 +/- 320 newtons); with PMMA, the yield strength increased by 26.8 percent (from 1,477 +/- 526 newtons to 1,834 +/- 225 newtons). However, only the increase with PMMA augmentation was significant at p < 0.05). The energy to yield increased significantly (41 percent, p < 0.05) with both types of augmentation (from 2,399 +/- 1,186 newton-millimeters to 3,378 +/- 857 newton-millimeters for calcium phosphate cement, and from 2,635 +/- 1,113 newton-millimeters to 3,741 +/- 426 newton-millimeters for PMMA), whereas the stiffness increased only slightly with PMMA augmentation (6.2 percent, from 481 +/- 180 newtons per millimeter to 511 +/ 92 newtons per millimeter) and fell slightly with calcium phosphate cement augmentation (10 percent, from 457 +/- 201 newtons per millimeter to 411 +/- 663 newtons per millimeter). CONCLUSIONS: The in situ-setting calcium phosphate cement investigated in this study compared favorably with PMMA in a single-cycle cut-out test of augmented compression hip screws in senile trabecular bone. Our results suggest that these materials may have promise as substitutes for PMMA in the salvage of compression hip screw fixation in elderly osteopenic patients with complex intertrochanteric fractures and that further study of their use in this application is warranted. PMID- 9415865 TI - Iliosacral screw fixation: early complications of the percutaneous technique. AB - OBJECTIVE: To report on the early complications related to the percutaneous placement of iliosacral screws for the operative treatment of displaced posterior pelvic ring disruptions. STUDY DESIGN: Prospective, consecutive. SETTING: Level one trauma center. PATIENTS: One hundred seventy-seven consecutive patients with unstable pelvic ring fractures. One hundred two male and seventy-five female patients ranging in age from eleven to seventy-eight years (mean, thirty-two years). INTERVENTIONS: Operative procedures were performed urgently according to the patient's clinical condition. Anterior pelvic reductions and fixations were performed by using internal and external fixation techniques. Accurate closed or open reductions of the posterior pelvic ring disruptions were accomplished by using a variety of surgical techniques dependent on the specific pattern of pelvic ring disruption. Closed manipulative reductions of the posterior pelvic ring were attempted for all patients. Open reductions were necessary in those patients with unacceptable closed manipulative reductions as assessed fluoroscopically at the time of operation (more than one centimeter in any field of fluoroscopic imaging). MAIN OUTCOME MEASURES: Plain inlet and outlet radiographs were obtained postoperatively at six weeks, three months, and twelve months. A pelvic computed tomography scan was performed postoperatively to assess fracture or dislocation reduction and the implant safety. Annual follow-up pelvic radiographs were obtained. Residual pelvic deformities were quantified based on these imaging modalities. RESULTS: There were no posterior pelvic infections. Minimal blood loss was associated with this technique. Complications occurred due to inadequate imaging, surgeon error, and fixation failure. Fluoroscopic imaging was inadequate due to obesity or abdominal contrast in eighteen patients. Five screws were misplaced due to surgeon error. One misplaced screw produced a transient L5 neuropraxia. Fixation failures related to either crandiocerebral trauma, delayed union, noncomplicance, and a deep anterior pelvic polymicrobial infection secondary to a urethral tear occurred in seven patients. There were two sacral nonunions that required debridement, bone grafting, and repeat fixation prior to healing. CONCLUSIONS: Iliosacral screw fixation of the posterior pelvis is difficult. The surgeon must understand the variability of sacral anatomy. Quality triplanar fluoroscopic imaging of the accurately reduced posterior pelvic ring should allow for safe iliosacral screw insertions. Anticipated noncompliant patients or those with craniocerebral trauma may need supplementary posterior pelvic fixation. Low rates of infection, blood loss, and nonunion can be expected. PMID- 9415866 TI - Stability of open-book pelvic fractures using a new biomechanical model of single limb stance. AB - OBJECTIVE: A new biomechanical model of single-limb stance was developed to test the stability of intact, injured, and internally fixed pelves. DESIGN: Single limb stance was simulated by applying muscle forces and body mass loading to cadaver pelves. We created a rotationally unstable "open-book" pelvic injury in nine embalmed pelves by dividing the ligaments of the pubic symphysis, pelvic floor, and anterior and interosseus sacroiliac joint. All pelves were devoid of gross structural abnormalities. INTERVENTION: Two methods of internal fixation of the pubis symphysis were compared: (a) a curved six-hole 3.5-millimeter reconstruction plate across the superior pubic symphysis, and (b) the same six hole 3.5-millimeter reconstruction plate plus a perpendicularly oriented four hole 3.5-millimeter reconstruction plate placed across the anterior symphysis. MAIN OUTCOME MEASUREMENTS: We measured vertical shear displacement at the public symphysis and horizontal displacement at the anterior sacroiliac joint. The results for the injured and fixed specimens were compared with each other and with the results for the intact specimens. RESULTS: The injured unfixed specimens showed marked instability that was prevented by both methods of fixation of the pubic symphysis. No significant differences could be demonstrated between single and double plating of the disrupted pubic symphysis when using this single-limb stance model. CONCLUSION: This model of single-limb stance suggests that a single symphyseal plate across the pubic symphysis can stabilize the open-book injury under short-term quasi-static loads. PMID- 9415867 TI - Functional recovery following hip fracture in the elderly. PMID- 9415868 TI - Food foments European disunity. PMID- 9415869 TI - The yeast genome yields its secrets. PMID- 9415870 TI - The role of innovation in drug development. PMID- 9415871 TI - Behind the color code of "no". PMID- 9415872 TI - Recombinant approaches for accessing biodiversity. PMID- 9415873 TI - Insect resistance to Bt revisited. PMID- 9415874 TI - Paradigm lost? PMID- 9415875 TI - Euro indecision triggers food anarchy. PMID- 9415876 TI - Firms sleuth out transgenic foods. PMID- 9415877 TI - Rhone-Poulenc rounds up DeKalb and Monsanto. PMID- 9415878 TI - Minicrash tests biotechnology investors and companies as market fluctuates. PMID- 9415879 TI - Amgen hangs hopes on hepatitis C. PMID- 9415880 TI - "NIH-money-for-drug-patent-life" proposal rejected. PMID- 9415881 TI - Complete genome expression monitoring: the human race. PMID- 9415882 TI - Seedless hopes bode well for winter vegetables. PMID- 9415883 TI - Spectrometry senses more than a small difference. PMID- 9415884 TI - A systems perspective begins to emerge in Alzheimer's. PMID- 9415885 TI - Shock toxicology with transgenics. PMID- 9415886 TI - Stress tolerance: the key to effective strains of industrial baker's yeast. AB - Application of yeasts in traditional biotechnologies such as baking, brewing, distiller's fermentations, and wine making, involves them in exposure to numerous environmental stresses. These can be encountered in concert and sequentially. Yeast exhibit a complex array of stress responses when under conditions that are less than physiologically ideal. These responses involve aspects of cell sensing, signal transduction, transcriptional and posttranslational control, protein targeting to organelles, accumulation of protectants, and activity of repair functions. The efficiency of these processes in a given yeast strain determines its robustness, and to a large extent, whether it is able to perform to necessary commercial standards in industrial processes. This article reviews aspects of stress and stress response in the context of baker's yeast manufacturing and applications, and discusses the potential for improving the general robustness of industrial baker's yeast strains, in relation to physiological and genetic manipulations. PMID- 9415887 TI - Genome-wide expression monitoring in Saccharomyces cerevisiae. AB - The genomic sequence of the budding yeast Saccharomyces cerevisiae has been used to design and synthesize high-density oligonucleotide arrays for monitoring the expression levels of nearly all yeast genes. This direct and highly parallel approach involves the hybridization of total mRNA populations to a set of four arrays that contain a total of more than 260,000 specifically chosen oligonucleotides synthesized in situ using light-directed combinatorial chemistry. The measurements are quantitative, sensitive, specific, and reproducible. Expression levels ranging from less than 0.1 copies to several hundred copies per cell have been measured for cells grown in rich and minimal media. Nearly 90% of all yeast mRNAs are observed to be present under both conditions, with approximately 50% present at levels between 0.1 and 1 copy per cell. Many of the genes observed to be differentially expressed under these conditions are expected, but large differences are also observed for many previously uncharacterized genes. PMID- 9415888 TI - Genetic analysis by peptide nucleic acid affinity MALDI-TOF mass spectrometry. AB - The ability to analyze multiple polymorphic sites rapidly and accurately is crucial in all areas of genetic analysis. We have developed an approach for the detection of multiple point mutations, using allele-specific, mass-labeled, peptide nucleic acid (PNA) hybridization probes, and direct analysis by matrix assisted laser desorption/ionization time-of-flight mass spectrometry. The composite mass spectra produced contain peaks of distinct masses corresponding to each allele present, resulting in a mass spectral "fingerprint" for each DNA sample. The hybridization characteristics of PNA:DNA duplexes were found to be highly dependent on both base content and sequence. Results from the analysis of four polymorphic sites contained in exon 4 of the human tyrosinase gene show that this approach is simple, rapid, and accurate with potential applications in many areas of genetic analysis. PMID- 9415890 TI - The endocrine secretion of human insulin and growth hormone by exocrine glands of the gastrointestinal tract. AB - The exocrine pancreas, liver, and submandibular glands of the rat were used to express and secrete two exogenous, human protein hormones (growth hormone and insulin) into blood at physiological concentrations. Transfection, expression, and secretion were achieved by the in vivo retrograde injection of plasmid DNA into the secretory ducts of these glands. Pancreatic acinar cells secreted physiological concentrations of growth hormone into the circulation, and its secretion was enhanced by cholinergic stimulation. A human insulin gene was engineered to allow normal processing of insulin in non-beta cells. With this gene, the secretion of human insulin by the exocrine pancreas normalized elevated blood glucose levels in diabetic rats. These in vivo observations demonstrate the utility of retrograde ductal administration of naked DNA into exocrine organs as a novel method for the regulated systemic delivery of protein-based pharmaceuticals. PMID- 9415889 TI - A cell surface tethered enzyme improves efficiency in gene-directed enzyme prodrug therapy. AB - The potential for expressing the bacterial enzyme carboxypeptidase G2 (CPG2) tethered to the outer surface of mammalian cells was examined for use in gene directed enzyme prodrug therapy. The affinity of CPG2 for the substrate methotrexate was unaffected by three mutations required to prevent N-linked glycosylation. Breast carcinoma MDA MB 361 cells expressing CPG2 internally showed only a very modest increase in sensitivity to the prodrug CMDA because the prodrug did not enter the cells. Cells expressing surface-tethered CPG2, however, became 16-24-fold more sensitive to CMDA and could mount a good bystander effect. Systemic administration of CMDA to mice bearing established xenografts of the transfected cells led to sustained tumor regressions or cures. PMID- 9415891 TI - Adenoviral preterminal protein stabilizes mini-adenoviral genomes in vitro and in vivo. AB - In the absence of host immunity, nonintegrating, first-generation adenoviral vectors remain stable in the nucleus of quiescent transduced cells in mice. A mini-adenoviral genome (9 kb) deleted for viral E1, E2, E3, and late genes, but containing the viral inverted terminal repeats (ITRs), transgene expression cassette (human alpha 1-antitrypsin), and the viral E4 genes was equally efficient at transducing cells in vitro or in vivo as first generation, E1 deleted vectors. In contrast to a first generation vector, gene expression as well as vector DNA was short-lived in cells transduced with the deleted adenoviral genome. We demonstrate that coexpression of the adenoviral E2 preterminal protein from the vector or in trans stabilizes the mini-genome in vitro and in vivo without evidence of cellular toxicity. PMID- 9415892 TI - Sustainable cutaneous gene delivery. AB - Durable gene delivery to human skin is necessary for lasting correction of human genetic skin disease. Current cutaneous gene-delivery strategies, however, have achieved only transient gene expression, often only within a small percentage of tissue cells. The recent inability to sustain phenotypic correction of human genetic skin disease due to loss of therapeutic gene expression in regenerated epidermal tissue has highlighted this current limitation. In an effort to surmount this problem, we have generated gene delivery vectors that produce more durable gene delivery in human skin tissue in vivo. PMID- 9415893 TI - A transgenic mouse model for the detection of cellular stress induced by toxic inorganic compounds. AB - Transgenic mice for genotoxicity testing have been developed, although no such models have been produced for the evaluation of toxic, nongenotoxic chemical compounds. We have developed a transgenic mouse model for the analysis of toxic inorganic compounds. We engineered a mouse lineage with the human growth hormone (hGH) gene under the control of the human hsp70 promoter, in which a plasma detectable hGH response can be elicited by exposure to heat shock. In primary cell cultures from these mice, hGH release was observed following treatment with several toxic inorganics. Transgenic mice injected intraperitoneally with sodium arsenite, cadmium chloride, copper sulphate, or methylmercurium chloride showed significant hGH levels in plasma. PMID- 9415894 TI - Genetic engineering of parthenocarpic plants. AB - Transgenic tobacco and eggplants expressing the coding region of the iaaM gene from Pseudomonas syringae pv. savastanoi, under the control of the regulatory sequences of the ovule-specific DefH9 gene from Antirrhinum majus, showed parthenocarpic fruit development. Expression of the DefH9-iaaM chimeric transgene occurs during flower development in both tobacco and eggplant. Seedless fruits were produced by emasculated flowers. When pollinated, the parthenocarpic plants produced fruits containing seeds. In eggplant, the genetic manipulation allowed fruit set and growth under environmental conditions prohibitive for fruit setting in the untransformed line, which did not set fruit at all. Under normal environmental conditions, production of marketable fruits took place from pollinated and unpollinated transgenic flowers, while flowers of untransformed control plants did produce fruits of marketable size only from fertilized flowers. PMID- 9415895 TI - As a matter of scientific fact: applying the inherency doctrine in biotechnology. PMID- 9415896 TI - Microbial biocatalysis and biodegradation informatics. PMID- 9415897 TI - Biotechnologies to watch. PMID- 9415898 TI - Neurotransmitters regulating defensive rage behavior in the cat. AB - This review summarizes recent findings of our laboratory that have been directed at: (1) identifying the neural circuits underlying the expression and modulation of defensive rage behavior in the cat and the neurotransmitters associated with these pathways; and (2) determining which components of the circuitry are affected by alcohol administration and which significantly alter the rage mechanism. The experiments described herein incorporated a number of converging methods, which include brain stimulation, behavioral pharmacology, immunocytochemistry, retrograde tract tracing and receptor binding. For behavioral pharmacological studies, monopolar electrodes and cannula-electrodes were implanted into selected regions along the limbic-midbrain axis for electrical stimulation and local microinfusion of drugs. The findings demonstrated: (1) a direct pathway from the anterior medial hypothalamus to the dorsal periaqueductal gray (PAG) over which this response is mediated. This pathway utilizes excitatory amino acids that act upon NMDA receptors within the midbrain PAG; (2) that the region of the dorsal PAG, from which defensive rage could be elicited, receives other inputs from the basal amygdala that facilitate this response by acting upon NMDA receptors; (3) a pathway from the medial amygdala to the medial hypothalamus that also facilitates defensive rage and whose functions are mediated by substance P receptors within the medial hypothalamus; (4) that the PAG also receives enkephalinergic inputs from the central nucleus of amygdala, which act upon mu receptors, and which powerfully suppress defensive rage; and (5) that recent findings reveal that ethanol administration facilitates defensive rage by virtue of its interactions with the medial hypothalamus, its descending projection to the PAG, and possibly with NMDA receptors within this pathway. PMID- 9415899 TI - Opposing roles of the amygdala and dorsolateral periaqueductal gray in fear potentiated startle. AB - The whole-body acoustic startle response is a short-latency reflex mediated by a relatively simple neural circuit in the lower brainstem and spinal cord. The amplitude of this reflex is markedly enhanced by moderate fear levels, and less effectively increased by higher fear levels. Extensive evidence indicates that the amygdala plays a key role in the potentiation of startle by moderate fear. More recent evidence suggests that the periaqueductal gray is involved in the loss of potentiated startle at higher levels of fear. The influence of both structures may be mediated by anatomical connections with the acoustic startle circuit, perhaps at the level of the nucleus reticularis pontis caudalis. The present chapter reviews these data. PMID- 9415900 TI - Transmitter systems involved in neural plasticity underlying increased anxiety and defense--implications for understanding anxiety following traumatic stress. AB - Lasting changes in anxiety-like behavior (ALB) may be produced in several ways. These include partial limbic kindling, injection of the beta-carboline FG-7142, and brief, non-injurious, exposure of rodents to cats (predator stress). Both seizures and FG-7142 induce long-term potentiation (LTP) in efferent pathways of the amygdala known to participate in feline defensive behavior. By comparing the behavioral and physiological effects of partial kindling and injection of FG 7142, NMDA-dependent LTP in the right amygdalo-periacqueductal gray (PAG) pathway emerges as being critical to maintained increases in feline ALB. A similar dependence on NMDA-mediated processes is described for lasting increases in rodent ALB following predator stress. The lasting aftereffects of predator stress on a variety of measures parallel many of the symptoms of post-traumatic stress disorder (PTSD). Support is provided for the idea that behavioral changes following FG-7142 and predator stress may model anxiety associated with PTSD. Moreover, it is suggested that both models share mechanisms in common involving the PAG. These mechanisms likely involve initiation of LTP by NMDA receptors, and prolongation of LTP by CCKB receptors. To the extent that response to the stressors reviewed here mimics the symptoms of PTSD, the data implicate NMDA mediated processes in the creation of what van der Kolk has called permanent emotional memories in PTSD. Their representation may be in the form of NMDA dependent LTP of transmission within the amygdala and between the amygdala and its efferents. CCK may play a pivotal role in prolonging limbic LTP and anxiety following traumatic stress. Since block of CCKB receptors before and after the stressor prevents lasting increases in ALB, pharmacological intervention to block CCK receptors shortly after a traumatic stressor might be efficacious in mitigating the permanence of these emotional memories. PMID- 9415901 TI - Anxiogenic-like consequences in animal models of complex partial seizures. AB - Several kinds of psychiatric symptoms (anxiety, depression, schizophrenia) have been associated with epilepsies, and clinical data suggest that patients with seizures involving limbic structures are the most prone to develop behavioural disorders between the seizures (i.e. interictally). Studying the neurobiological mechanisms that underlie these symptoms is difficult in humans because of different interfering factors (e.g. psychosocial difficulties, pharmacological side-effects, lesions), which can be avoided in animal models. Using repetitive electrical stimulations (kindling) or local applications of a neuroexcitotoxin in limbic structures (mainly the amygdala and hippocampus), several authors have reported lasting changes of emotional reactivity in cats and rats. These changes appear as anxiety-related reactions expressed as a hyperdefensiveness in the cat, or a reduction of spontaneous exploration in tests predictive of anxiogenic effects in the rat. Some neuroplasticity processes known to develop during epileptogenesis (neuronal-hyperexcitability, modulation of GABA/benzodiazepine transmission) may participate in these lasting changes of behaviour, especially in structures involved in the control of fear-promoted reactions (amygdala, periaqueductal grey matter). In addition, endogenous control systems may also play a critical role in the occurrence of interictal behavioural disorders. PMID- 9415902 TI - The temporal dynamics of the stress response. AB - This paper summarises the available evidence that failure of defense mechanisms in (semi)-natural social groups of animals may lead to serious forms of stress pathology. Hence the study of social stress may provide animal models with a high face validity. However, most of the animal models of human stress-disorders have concentrated on the consequences of chronic exposure to stressors. The present paper considers recent data, indicating that a single experience with a major stressor in the form of social defeat may have long-term consequences ranging from hours to days and weeks. It seems that the experience of a major stressor sensitizes the animal to subsequent stressors. The consequences of these long term temporal dynamics of the stress response to the development of stress related disorders and stress-vulnerability are discussed. PMID- 9415903 TI - Differentiation of anxiolytic and panicolytic drugs by effects on rat and mouse defense test batteries. AB - The use of ethoexperimental techniques to elicit and maximize the full range of defensive behaviors of rats and mice enables a very precise analysis of the effects of drugs on these behavior patterns. Two rat defense test batteries (the fear/defense test battery or F/DTB and the anxiety/defense test battery or A/DTB) have provided evidence that anxiolytic drugs, even from different classes, produce a common pattern of changes in specific behaviors. A recently developed mouse defense test battery (MDTB) has enabled description of mouse defensive behaviors to a predator, for comparison to those of rats, and a series of studies of drug effects on the behaviors measured in the MDTB provides evidence of cross species generality of anxiolytic drug effects, or lack of effect, on specific defensive behaviors. In addition, tests with panicogenic and panicolytic drugs in the MDTB indicate that these enhance and reduce, respectively, flight reactions, which generally are not altered by anxiolytic compounds. Thus, results from the MDTB, taken in conjunction with those of the two rat test batteries and other defense analyses in rats and mice, provide evidence that many defensive behaviors are similar across rodent species, while the differences obtained provide a consistent pattern across situations. Moreover, the defense test batteries may be used to differentiate the effects of drugs effective against generalized anxiety as opposed to panic, through effects on specific defensive behaviors. PMID- 9415904 TI - Dual role of 5-HT in defense and anxiety. AB - Pharmacological results obtained in animals tested in approach/avoidance conflict situations have led to the suggestion that 5-HT enhances anxiety by acting on forebrain structures. In contrast, results with intracerebral drug injection associated with aversive electrical brain stimulation indicate that 5-HT inhibits aversion in the dorsal periaqueductal gray (DPAG). To reconcile this evidence, it has been suggested that 5-HT may enhance conditioned fear in the amygdala while inhibiting innate fear in the DPAG. To test this hypothesis, we used three drug treatments known to increase the release of 5-HT from terminals of the dorsal raphe nucleus (DR): (1) intra-DR microinjection of the benzodiazepine inverse agonist FG 7142, (2) intra-DR microinjection of the excitatory amino acid kainic acid and (3) intraperitoneal injection of the 5-HT releaser and uptake blocker D fenfluramine. All drug treatments enhanced inhibitory avoidance (learned fear) in the elevated T-maze, a new animal model of anxiety. Intra-raphe kainate and D fenfluramine also decreased one-way escape (innate fear) in the T-maze. In contrast, reduction of 5-HT release by intra-DR injection of 8-OH-DPAT impaired inhibitory avoidance without affecting one-way escape. Overall, these results agree with the above hypothesis. Clinical implications are discussed, especially with regard to panic disorder. PMID- 9415905 TI - Anxiety, defence and the elevated plus-maze. AB - The elevated plus-maze test has been in use as a rodent model of anxiety for a decade, and is representative of those tests that are based upon the study of spontaneous behaviour patterns and which have high ecological validity. The origins of the test in studies of the relationship between exploration and fear are reviewed, and attention is drawn to the distinct possibility that variation in the pharmacosensitivity of the procedure may be attributable to often extreme methodological variation between laboratories. In considering further this issue, attention is also drawn to the need to collect data under constant test conditions and to provide the minimum database necessary to reach conclusions regarding the behavioural specificity of drug action. Recent research, which has extended the conventional plus-maze scoring technique to include specific behavioural acts and postures (in particular, those relating to defensive behaviour), is described. The value of such an ethological approach to the plus maze is then exemplified with original data that demonstrate behaviourally selective, anti-anxiety effects of the GABAA receptor agonist, muscimol (0.125 1.0 mg/kg). It is concluded that, when used appropriately, the elevated plus-maze test can be a very valuable tool in drug screening and in the study of the neurobiology of anxiety and defence. More attention to behaviour and somewhat less emphasis on test simplicity and convenience would seem to be warranted. PMID- 9415906 TI - Managed care threatening health research. PMID- 9415908 TI - Isolated closed diaphyseal fractures of the femur in children: comparison of effectiveness and cost of several treatment methods. AB - The effectiveness of several treatment modalities for isolated closed femur fractures in children ages 4 through 16 years is compared based on outcome (clinical results, morbidity, and parents' satisfaction) and cost. Between 1986 and 1993, 30 patients were treated. Treatment methods included immediate hip spica cast application, distal femoral skeletal traction pin to align the fracture followed by early hip spica cast incorporating the pin (6th day), in hospital traction, primary external fixation, and primary intramedullary nailing. Average follow up was 4.1 years. Overall results were excellent with minimal morbidity for all treatment methods. Early application of hip spica cast with or without incorporation of the distal femoral traction pin required the fewest hospital days, the shortest time to union, and had the lowest overall cost. Complications, mainly malunion and femoral length discrepancy, however, were greater. Skeletal traction resulted in the longest hospital stay and was equal in cost to external fixation and intramedullary nailing. Primary external fixation appears most applicable in patients who are unlikely to tolerate early casting and who are at increased risk of avascular necrosis with femoral nailing. Patients treated with an intramedullary nail had the fewest complications. Age, size, and gender of patient, location and type of fracture, as well as social circumstances are all significant factors in choosing the optimal method of treatment. PMID- 9415907 TI - Comparison of closed-suction drainage and no drainage after primary total knee arthroplasty. AB - One hundred thirty-six primary total knee arthroplasty patients were randomized for the use of closed-suction, nonreinfusable wound drains. Blood loss was identical in the drained and undrained groups. Forty percent of undrained wounds compared with 0% of drained wounds required dressing reinforcement. Sixty-nine percent of undrained wounds compared with 39% of drained wounds developed ecchymosis, measuring 92 cm2 in the undrained group and 28 cm2 in the drained group. This study concludes that a simple wound drain effectively minimizes the undesirable accumulation of blood in the surrounding soft tissues and the postoperative wound dressing after total knee arthroplasty. PMID- 9415909 TI - Tourniquet pressures on pediatric patients: a clinical study. AB - A clinical study was undertaken to evaluate pneumatic tourniquet pressures required for hemostasis in extremity surgery of pediatric patients. Occlusion pressures were measured by Doppler, and tourniquet pressures were set 50 mm above this value. Of 29 cases, 86% were determined to provide adequate hemostasis throughout the procedure. Maximum mean pressures used in the upper and lower extremity groups were 173.4 +/- 11.6 mm Hg (range: 155 to 190 mm Hg) and 176.7 +/ 28.7 mm Hg (range: 140 to 250 mm Hg), respectively, accounting for adjustments made to inadequate initial settings. This study suggests that lower tourniquet pressures than previously used may be needed to maintain adequate hemostasis in pediatric patients. PMID- 9415910 TI - Complications of CT-guided biopsy of the spine and sacrum. AB - This study reviews the results of 94 computed tomography (CT)-guided Craig needle biopsies of the spine and sacrum performed at one center. An indication for biopsy in this study was prompted by abnormal findings identified by one or more of the following diagnostic modalities: radiography, CT, magnetic resonance imaging (MRI), or bone scanning. These patients then underwent CT-guided Craig needle biopsy of the spine and sacrum for further evaluation. There were 1 biopsy of the cervical spine, 19 of the thoracic spine, 66 of the lumbar spine, and 8 of the sacrum. Biopsy sensitivity was 94.5% and specificity was 96.8%. This accuracy compared with other diagnostic modalities showed biopsy to be the gold standard for diagnosis of spine or sacral lesions. Of the 94 cases reviewed, 6 complications were noted. All complications were acute in nature and included 1 aortic puncture, 2 psoas punctures with associated psoas hematomas, 1 biopsy of an incorrect level, and 2 aborted procedures secondary to patient discomfort. No infections or neurological sequelae were seen. Although the benefits of CT-guided biopsy over open biopsy have been shown previously, this review demonstrates it is not without significant risk. PMID- 9415911 TI - Effect and cost of subcutaneous recombinant human erythropoietin in preoperative patients. PMID- 9415912 TI - The theory of early loosening of hip prostheses. AB - The issue of prosthetic loosening is currently a matter of debate, particularly with regard to the timing and nature of the precipitating events. The theory presented here postulates that loosening begins at an early stage due to either insufficient initial fixation or early loss of fixation. The loosened prosthetic component is then affected by varying degrees of mechanical stress associated with normal daily activity, which differs according to patient characteristics (body weight and level of physical activity) and the components used (prosthetic design, positioning, friction, and wear). This theory of early loosening can explain without supplementary ad hoc assumptions the rapid early prosthetic migration detected by roentgen stereophotogrammetry, the development of focal osteolysis and wear granulomas, the phenomenon known as stress-shielding, and, to a great extent, the epidemiology of clinical failure. PMID- 9415913 TI - Dissociation of a polyethylene liner in a nonmodular, cemented, metal-backed acetabular component. PMID- 9415914 TI - Simultaneous bilateral glenoid fractures associated with glenohumeral subluxation/dislocation in a weightlifter. PMID- 9415915 TI - Glenohumeral pyarthrosis following acupuncture treatment. PMID- 9415916 TI - Bizarre parosteal osteochondromatous proliferation of bone. PMID- 9415917 TI - Structure and function of the Nautilus statocyst. AB - The two equilibrium receptor organs (statocysts) of Nautilus are avoid sacks, half-filled with numerous small, free-moving statoconia and half with endolymph. The inner surface of each statocyst is lined with 130,000-150,000 primary sensory hair cells. The hair cells are of two morphological types. Type A hair cells carry 10-15 kinocilia arranged in a single ciliary row; they are present in the ventral half of the statocyst. Type B hair cells carry 8-10 irregularly arranged kinocilia; they are present in the dorsal half of the statocyst. Both type of hair cells are morphologically polarized. To test whether these features allow the Nautilus statocyst to sense angular accelerations, behavioural experiments were performed to measure statocyst-dependent funnel movements during sinusoidal oscillations of restrained Nautilus around a vertical body axis. Such dynamic rotatory stimulation caused horizontal phase-locked movements of the funnel. The funnel movements were either in the same direction (compensatory funnel response), or in the opposite direction (funnel follow response) to that of the applied rotation. Compensatory funnel movements were also seen during optokinetic stimulation (with a black and white stripe pattern) and during stimulations in which optokinetic and statocyst stimulations were combined. These morphological and behavioural findings show that the statocysts of Nautilus, in addition to their function as gravity receptor organs, are able to detect rotatory movements (angular accelerations) without the specialized receptor systems (crista/cupula systems) that are found in the statocysts of coleoid cephalopods. The findings further indicate that both statocyst and visual inputs control compensatory funnel movements. PMID- 9415918 TI - Amnesia, memory and brain systems. AB - Bilateral damage to either the medial temporal lobe or the diencephalic midline causes an amnesic syndrome, i.e. a global impairment in the ability to acquire new memories regardless of sensory modality, and a loss of some memories, especially recent ones, from the period before amnesia began. The memory deficit can occur against a background of intact intellectual and perceptual functions. Two themes have been prominent in recent work. First, the amnesic syndrome is narrower than once believed in the sense that a number of learning and memory abilities are preserved (e.g. skill and habit learning, simple forms of conditioning and the phenomenon of priming). Second, the brain system damaged in amnesia has only a temporary role in memory. As time passes after learning, memory is reorganized and consolidated within neocortex, such that eventually medial temporal lobe and diencephalic structures are not needed for storage or retrieval. PMID- 9415919 TI - Amygdala and bed nucleus of the stria terminalis: differential roles in fear and anxiety measured with the acoustic startle reflex. AB - Neural stimuli associated with traumatic events can readily become conditioned so as to reinstate the memory of the original trauma. These conditioned fear responses can last a lifetime and may be especially resistant to extinction. A large amount of data from many different laboratories indicate that the amygdala plays a crucial role in conditioned fear. The amygdala receives information from all sensory modalities and projects to a variety of hypothalamic and brainstem target areas known to be critically involved in specific signs that are used to define fear and anxiety. Electrical stimulation of the amygdala elicits a pattern of behaviours that mimic natural or conditioned states of fear. Lesions of the amygdala block innate or conditioned fear and local infusion of drugs into the amygdala have anxiolytic effects in several behavioural tests. Excitatory amino acid receptors in the amygdala are critical for the acquisition, expression and extinction of conditioned fear. PMID- 9415920 TI - The cognitive neuroscience of memory: perspectives from neuroimaging research. AB - Cognitive neuroscience approaches to memory attempt to elucidate the brain processes and systems that are involved in different forms of memory and learning. This paper examines recent research from brain-damaged patients and neuroimaging studies that bears on the distinction between explicit and implicit forms of memory. Explicit memory refers to conscious recollection of previous experiences, whereas implicit memory refers to the non-conscious effects of past experiences on subsequent performance and behaviour. Converging evidence suggests that an implicit form of memory known as priming is associated with changes in posterior cortical regions that are involved in perceptual processing; some of the same regions may contribute to explicit memory. The hippocampal formation and prefrontal cortex also play important roles in explicit memory. Evidence is presented from recent PET scanning studies that suggests that frontal regions are associated with intentional strategic efforts to retrieve recent experiences, whereas the hippocampal formation is associated with some aspect of the actual recollection of an event. PMID- 9415921 TI - The ecological study of memory. AB - The study of memory has long been dominated by the structural tradition, and especially by the experimental analysis of mechanisms of information processing. That dominance may soon be brought to an end by the progress of neuroscience, which offers more direct ways of studying the mechanisms in question. At that point functional issues may move to centre stage. Those issues include the act of remembering and its social functions, the skills and presuppositions of the remembered, the interaction of those skills and presuppositions with the particular material being remembered, and the determinants of accuracy and confabulation in recall. PMID- 9415922 TI - The ageing brain: normal and abnormal memory. AB - With advancing age, the majority of individuals experience declines in their ability to learn and remember. An examination of brain structure and function in healthy older persons across the age range indicates that there are substantial changes in the brain that appear to be related to alterations in memory. The nature of the cognitive and neurobiological alterations associated with age related change is substantially different from that seen in the early stages of a dementing illness, such as Alzheimer's disease. These differences have implications for potential intervention strategies. PMID- 9415923 TI - Animal models of memory impairment. AB - Memory impairment in the elderly resembles a mild temporal lobe dysfunction. Alterations in the hippocampal formation are also a probable basis for cognitive deficits in some animal models of ageing. For example, aged rats are impaired in hippocampal-dependent tests of spatial memory. Recent studies have revealed considerable structural integrity in the aged hippocampus, even in aged rats with the most impaired spatial memory. In contrast, atrophy/loss of cholinergic neurons in the basal forebrain and deficiency in cholinergic transduction in hippocampus correlate with the severity of spatial memory impairment in aged rats. This evidence supports the longstanding view that age-related loss of memory has a cholinergic basis. In this context, it is somewhat surprising that the use of a selective cholinergic immunotoxin in young rats to further test this hypothesis has revealed normal spatial memory after removing septo-hippocampal cholinergic neurons. Young rats with immunotoxic lesions, however, have other behavioural impairments in tests of attentional processing. These lines of research have implications for understanding the neurobiological basis of memory deficits in ageing and for selecting an optimal behavioural setting in which to examine therapies aimed at restoring neurobiological function. PMID- 9415925 TI - Hypnosis, memory and amnesia. AB - Hypnotized subjects respond to suggestions from the hypnotist for imaginative experiences involving alterations in perception and memory. Individual differences in hypnotizability are only weakly related to other forms of suggestibility. Neuropsychological speculations about hypnosis focus on the right hemisphere and/or the frontal lobes. Posthypnotic amnesia refers to subjects' difficulty in remembering, after hypnosis, the events and experiences that transpired while they were hypnotized. Posthypnotic amnesia is not an instance of state-dependent memory, but it does seem to involve a disruption of retrieval processes similar to the functional amnesias observed in clinical dissociative disorders. Implicit memory, however, is largely spared, and may underlie subjects' ability to recognize events that they cannot recall. Hypnotic hypermnesia refers to improved memory for past events. However, such improvements are illusory: hypermnesia suggestions increase false recollection, as well as subjects' confidence in both true and false memories. Hypnotic age regression can be subjectively compelling, but does not involve the ablation of adult memory, or the reinstatement of childlike modes of mental functioning, or the revivification of memory. The clinical and forensic use of hypermnesia and age regression to enhance memory in patients, victims and witnesses (e.g. recovered memory therapy for child sexual abuse) should be discouraged. PMID- 9415924 TI - Emotional memory and psychopathology. AB - A leading model for studying how the brain forms memories about unpleasant experiences is fear conditioning. A cumulative body of work has identified major components of the neural system mediating this form of learning. The pathways involve transmission of sensory information from processing areas in the thalamus and cortex to the amygdala. The amygdala's lateral nucleus receives and integrates the sensory inputs from the thalamic and cortical areas, and the central nucleus provides the interface with motor systems controlling specific fear responses in various modalities (behavioural, autonomic, endocrine). Internal connections within the amygdala allow the lateral and central nuclei to communicate. Recent studies have begun to identify some sites of plasticity in the circuitry and the cellular mechanisms involved in fear conditioning. Through studies of fear conditioning, our understanding of emotional memory is being taken to the level of cells and synapses in the brain. Advances in understanding emotional memory hold out the possibility that emotional disorders may be better defined and treatment improved. PMID- 9415926 TI - Source monitoring and memory distortion. AB - Memory distortion reflects failures to identify the sources of mental experience (reality monitoring failures or source misattributions). For example, people sometimes confuse what they inferred or imagined and what actually happened, what they saw and what was suggested to them, one person's actions and another's what they heard and what they previously knew, and fiction and fact. Source confusions arise because activated information is incomplete or ambiguous and the evaluative processes responsible for attributing information to sources are imperfect. Both accurate and inaccurate source attributions result from heuristic processes and more reflectively complex processes that evaluate a mental experience for various qualities such as amount and type of perceptual, contextual, affective, semantic and cognitive detail, that retrieve additional supporting or disconfirming evidence, and that evaluate plausibility and consistency given general knowledge, schemes, biases and goals. Experimental and clinical evidence regarding cognitive mechanisms and underlying brain structures of source monitoring are discussed. PMID- 9415927 TI - Transient global amnesia and functional retrograde amnesia: contrasting examples of episodic memory loss. AB - We studied 11 patients with transient global amnesia (TGA) and ten patients with functional retrograde amnesia (FRA). Patients with TGA had a uniform clinical picture: a severe, relatively isolated amnesic syndrome that started suddenly, persisted for 4-12 h, and then gradually improved to essentially normal over the next 12-24 h. During the episode, the patients had severe anterograde amnesia for verbal and non-verbal material and retrograde amnesia that typically covered at least two decades. Thirty hours to 42 days after the episode, the patients had recovered completely and performed normally on tests of anterograde and retrograde amnesia. By contrast, patients with FRA had a sudden onset of memory problems that were characterized by severe retrograde amnesia without associated anterograde amnesia and with a clinical presentation that otherwise varied considerably. The episodes persisted from several weeks to more than two years, and some of the patients had not recovered at the time of our last contact with them. The uniform clinical picture of TGA and the variable clinical picture of FRA presumably reflect their respective neurologic ('organic') and psychogenic ('non-organic') aetiologies. PMID- 9415928 TI - Memory and anxiety disorders. AB - Experimental psychopathologists have identified varying patterns in memory bias in people with depressive and anxiety disorders. Individuals suffering from depression tend to exhibit explicit memory deficits for positively-valanced material, and sometimes exhibit biases for retrieving negative self-relevant information as well. Most studies, however, provide scant evidence for implicit memory biases in depression. In contrast to depression, anxiety disorders are rarely associated with enhanced explicit memory for threat-related information (with the exception of panic disorder). Evidence for implicit memory biases for threat in these syndromes is mixed. After providing an overview of findings on memory abnormalities in depressive and anxiety disorders, data from several new studies bearing on posttraumatic stress disorder (PTSD) in Vietnam combat veterans and in women with histories of childhood sexual abuse are presented. Involving directed forgetting, implicit memory and autobiographical cueing paradigms, these experiments point to a pattern of abnormalities linked to PTSD rather than to trauma per se. PMID- 9415929 TI - Renal artery embolism. PMID- 9415930 TI - The SHR as a small animal model for radiocontrast renal failure. Relation of nephrotoxicity to animal's age, gender, strain, and dose of radiocontrast. AB - The male spontaneously hypertensive rat (SHR), as it ages, suffers many of the renal and cardiovascular complications that are recognized in humans as risk factors for radiocontrast (RC) agent induced renal failure (RF). Knowledge of this led us to test this strain of rats as a small animal model for RC-induced renal failure (RC-RF). Functional studies demonstrated a significant fall in GFR in the recovery period after RC administration. In addition, histopathologic evaluation of the kidneys was done in this study. Our results are based on assigning separate scale values to the histopathological evaluation of the (a) glomeruli, (b) tubules, (c) interstitium, and (d) arteries and arterioles of the kidneys. Saline (S) was administered to one group and the RC agent Hypaque-76 (diatrizoate meglumine sodium) to paired groups of 5-, 8-, 10-, 12-, and 14-month old male SHR. The results indicated that younger animals (5 and 8 months old) were resistant to the nephrotoxic effects of the RC, but developed susceptibility at 10 months of age, when spontaneous renal pathology became manifest. Both spontaneous renal pathology and RC-induced renal damage (RC-RD) increased as the animals aged. In addition, when the administered dose of RC was repeated after a short interval of only 6 h, the degree of RC-RD increased greatly. In parallel control studies of the influence of gender and strain on the response to RC in 12 month-old rats, neither hypertensive female SHR nor male normotensive Wistar Kyoto (WKY) rats demonstrated significant spontaneous renal pathology or the marked susceptibility to RC nephrotoxicity shown by their male SHR counterparts. This small animal model for RC-RD, the mature male SHR, has the distinct advantage that risk factors for RC-RD, similar to those characterized in humans for RC-RF, develop spontaneously without requiring any special treatment or surgical intervention. PMID- 9415931 TI - Transport of paraquat by a renal epithelial cell line, MDCK. AB - Transport of paraquat (PQ), a herbicidal cation, was previously investigated in a proximal (LLC-PK1), renal epithelial cell line using permeable collagen-coated filters. PQ was actively transported from the basolateral side via a cation transport system by the LLC-PK1 cells. In the present study, the transport of PQ was investigated in a distal renal epithelial cell line, MDCK. PQ was predominantly transported from the basolateral to apical (B to A) side. The basolateral transport of PQ in MDCK cells was not saturable with increasing concentrations and not energy dependent. The flux and uptake of PQ was much lower in the MDCK than LLC-PK1 cells. It is concluded that MDCK, a distal renal tubular cell line, does not have an active transport system for PQ. PMID- 9415932 TI - Beneficial effect of ETA receptor blockade in a rat model of radiocontrast induced nephropathy. AB - Endothelin has been implicated in the pathogenesis of acute renal failure associated with radiocontrast media. The current study was designed to determine the effect of endothelin receptor blockade in a model of radiocontrast-induced nephropathy. Inhibition of prostanoids and nitric oxide with indomethacin and L nitroarginine methyl ester (L-NAME) predisposes rats to the nephrotoxic effects of radiocontrast media and, therefore, has been proposed as a clinically relevant model. Separate groups of conscious rats were given the ETA receptor antagonist, A-127722 (3, 10, or 30 mg/kg), or water (1 mL/kg) by oral gavage. All rats were then given indomethacin (10 mg/kg), L-NAME (10 mg/kg), and the contrast agent, diatrizoate (6 mL/kg), by intravenous injection at 15-min intervals. Urine was collected for the subsequent 24 h by placing rats in metabolism cages. When indomethacin, L-NAME, and diatrizoate were administered without A-127722, rats displayed typical signs of renal failure; protein excretion was increased from a baseline of 13 +/- 2 to 33 +/- 8 mg/day (p < 0.05) and plasma creatinine increased from 0.52 +/- 0.01 to 0.62 +/- 0.04 mg/dL (p < 0.05). ETA receptor blockade prevented the rise in protein excretion and plasma creatinine in a dose dependent manner. In separate series of clearance experiments, A-127722 completely inhibited the renal effects of exogenous ET-1. These results suggest that endothelin plays a role in the hemodynamic events in this model and that ETA receptor antagonists should be investigated as potential therapeutic agents for radiocontrast-induced nephropathy. PMID- 9415933 TI - Chronic renal failure in India. AB - A prospective study of all new cases of chronic renal failure (CRF) including inservice referrals was done at our hospital over a period of 1 year from May 1994 to April 1995. The diagnosis of CRF was based on clinical, laboratory, and radiological features. Kidney biopsies were done when indicated. The patients were subdivided into various etiologic groups of primary renal disease according to standard criteria. There were a total of 835 cases of CRF with a median age of 43 years (range 10 days to 90 years); 67.8% of them were men. Glomerulonephritis (28.6%), diabetic nephropathy (23.2%), and interstitial nephritis (16.5%) were the most common causes of CRF, followed by obstructive nephropathy (6.4%), benign nephrosclerosis (4.1%), and polycystic kidney disease (2%). However, in patients more than 40 years of age, diabetic nephropathy was the most common cause (36.8%). The cause of CRF was unknown in 16.2% of the cases. One hundred twenty one patients (14.5%) had an acute deterioration of their underlying renal dysfunction at presentation. This was most commonly due to accelerated hypertension (26.1%), infection (22.4%), volume depletion (20.1%), and drugs (14.9%). Anti-inflammatory drugs were the most common drugs responsible for the acute decline in renal function. One year after their initial presentation, of the 512 patients (61.3%) with end stage renal disease, 12.5% had died, 17% had received a kidney allograft, 12.7% were on some form of maintenance dialysis, and 295 patients were lost to follow-up. Of the 323 patients with less severe illness, 7 died, 209 were on outpatient treatment, and 107 patients were lost to follow-up. We conclude that the pattern of CRF in India does not differ greatly from that in the developed countries. However, it carries a poorer prognosis due to late referral and limited availability and affordability of renal replacement therapy in India. PMID- 9415934 TI - Red cell ferritin, a marker of iron deficiency in hemodialysis patients. AB - Estimation of red cell ferritin (RCFer) may give a good indication of iron supply to the erythron and it may therefore be clinically useful for the detection of functional iron deficiency. In a cross-sectional study of hemodialysis patients on erythropoietin (EPO) therapy and regular oral iron we have compared the RCFer levels with conventional indicators of iron status. The patients studied, 19 female, 48 male, mean age 62 +/- 3.6 years (range 20-83 years) were characterized by the following mean parameters: aluminum 1.24 +/- 0.12 mumol/L, PTH 115.7 +/- 39 pg/mL, vitamin B12 626 +/- 71.2 ng/L, serum folate 18.8 +/- 2.2 micrograms/L, and hemoglobin 9.8 +/- 0.3 g/dL (range 7.3-12.4). The median serum ferritin (SF), RCFer, total iron binding capacity (TIBC), transferrin saturation (TS), and serum iron were 68 micrograms/L, 14.1 ag ferritin/red cell, 57 mumol/L, 20% and 11.5 mumol/L, respectively. Eleven patients had a reduced RCFer (< 7 ag ferritin/red cell), 5 had a SF of < 15 micrograms/L and 22 a TS of < 16%. The occurrence of functional iron deficiency was suggested by the presence of 10 subjects with reduced RCFer despite normal SF levels (15-240 micrograms/L). Four patients with reduced SF showed acceptable levels of RCFer, suggesting that some patients may maintain an adequate iron supply despite diminished iron stores. Despite oral iron therapy, a significant number of patients (63%) on regular hemodialysis remain relatively iron deficient with a serum ferritin of less than 100 micrograms/L. It has previously been proposed that oral iron provides adequate supplementation during increased demand caused by EPO stimulation. The present study has demonstrated overt iron deficiency in five subjects and suggests functional iron deficiency in a further seven (22% of total patients). We therefore conclude that oral iron therapy cannot maximize the response to EPO. PMID- 9415935 TI - Sensitivity and specificity of transferrin saturation and serum ferritin as markers of iron status after intravenous iron dextran in hemodialysis patients. AB - This study was designed to investigate the effect of intravenous (i.v.) iron dextran (i.d.) on hematocrit (Hct), transferrin saturation (TS), and serum ferritin (SF) in hemodialysis patients treated with a constant dose of erythropoietin (EPO). The sensitivity, specificity, and predictive values of SF and TS for monitoring i.d. therapy were also assessed. All hemodialysis patients with baseline SF < 100 ng/mL or TS < 20%, with EPO dose unchanged 6 weeks before and 4 weeks after dosing with i.d. were included. I.d. (500 mg-1 g) was given as an infusion over 1 h. Patients receiving packed RBC or with active bleeding were excluded. Hct, TS, and SF were measured 2 weeks before and 4 weeks after i.d. Linear correlation coefficients between dose of i.d., changes in Hct, TS, and SF were calculated. The sensitivity, specificity, and predictive values of TS and SF were compared. A positive Hct response was defined as a > 5% increase from baseline 4 weeks after administration of i.d. Thirty-three patients (17 females) received a total of 51 doses of i.d. Mean +/- SD i.d. dose was 770 +/- 278 mg. Hct increased by a mean +/- SD of 4.8% +/- 9.9% (33.4% +/- 3.0% to 34.9% +/- 4.1% [p = 0.028]); SF rose by a median of 208.65% (mean +/- SD of 126.8 +/- 132.1 ng/mL to 325.3 +/- 222.0 ng/mL [p < 0.0001]; TS increased by a median of 53.8% (19.4% +/- 9.4% to 29.3% +/- 11.3% [p < 0.0001]) from baseline values. The correlations between dose of ID and percent changes in SF, TS, and Hct were poor (r2 < 0.02). The sensitivities and specificities were 74% and 36% (TS < 20% alone); 60% and 30% (SF < 100 ng/mL alone); and 33% and 67% (TS < 20% and SF < 100 ng/mL), respectively. The predictive values for positive responses were 48% for TS and 45% for SF when used alone, and 47% when both indices were used together. The predictive value increased to 65% when either SF < 100 ng/mL or TS < 20% were used. At a constant EPO dose, there was a statistically significant increase in Hct 4 weeks after i.d. administration in patients who were diagnosed with iron deficiency by using TS < 20% or SF < 100 ng/mL. The dose of i.d. administered was poorly correlated to changes in Hct, TS, and SF. Both TS and SF are non-specific and insensitive indicators for accurate diagnosis of iron deficiency in hemodialysis patients in EPO. PMID- 9415936 TI - Assessment of dialysis adequacy using the dialysate urea monitor: preliminary experience of the dialysate urea monitor. AB - Numerous studies have identified a strong linkage between the delivered dialysis dose (Kt/V) and the survival of hemodialysis (HD) patients. However, the current method used to calculate Kt/V requires multiple blood samples and the process is complex and time consuming. We evaluate the performance of a recently developed on-line monitor (Biostat 1000 dialysate urea monitor, Baxter) that measures the urea concentration in the effluent dialysate and displays Kt/V and nPCR immediately after hemodialysis. To verify the performance of the urea monitor, we selected 21 hemodialysis patients, calculated their Kt/V and nPCR values from blood samples obtained during each hemodialysis, and compared the results with data obtained using the urea monitor. The Kt/V and nPCR values calculated by the urea monitor were both significantly correlated with those obtained using blood samples (R = 0.804, p < 0.001 in Kt/V and R = 0.749, p < 0.001 in nPCR). Our results suggest that the urea monitor may be used for on-line assessment of dialysis adequacy and obviates the need for blood sampling. PMID- 9415937 TI - Left ventricular morphology in chronic renal failure by echocardiography. AB - M-mode, two-dimensional, and Doppler echocardiography were performed in 38 chronic renal failure (CRD) patients on conservative management, 35 patients on hemodialysis, and 36 matched controls. The controls were matched for age, sex, and comorbidities. The incidence of hypertension, left ventricular (LV) end diastolic volume, LV end systolic volume, and LV mass index were significantly higher in patients on hemodialysis compared to the controls. The LV parameters in the predialysis patients were not significantly different from the controls, except the LV end systolic internal dimensions were significantly higher in the CRF patients. Multiple regression analysis underscored the strong association between increase in LV mass index (LVMI) and hypertension. The diabetic patients with renal failure had large LV internal diameter and end diastolic volume compared to non-diabetics. Systolic function was well preserved even in hypertensive and diabetic patients with uremia. The incidence of diastolic dysfunction and asymmetrical septal hypertrophy were not significantly different in the three groups of patients. PMID- 9415938 TI - Outcome of acute renal failure in meningococcemia. AB - We studied 28 consecutive patients (18 males and 10 females), 1-32 years of age, admitted to the intensive care unit from January 1989 to July 1995, with acute renal failure (ARF) due to meningococcal septicemia. All patients were treated with dexamethasone, penicillin, and/or chloramphenicol. Twenty-two patients presented septic shock and needed fluid replacement and vasoactive drugs. Acute renal failure was oliguric in 67.8%. Maximum levels of blood urea and serum creatinine were 210.3 +/- 26.6 mg/dL and 6.9 +/- 1.3 mg/dL, respectively. Metabolic acidosis was observed in 89.3% and hyperkalemia in 43%. The fractional excretion of sodium on day 1 was high (9.9 +/- 0.6%). The urinalysis did not show trace protein, but hematuria was positive in 81%. The mortality rate was 63.3%. In the 10 survivors, oliguria was present for a period of 12.7 +/- 2.4 days, and the period to reach a normal serum creatinine level was 20.2 +/- 4.7 days, although in two female patients, 7 and 17 years old, the elevated serum creatinine persisted. Renal biopsy was performed in one of these patients which revealed bilateral cortical necrosis. These data show that acute renal failure in meningococcemia presents high mortality rate associated to shock; 80% of the survivors recover renal function; and bilateral cortical necrosis occurred in one patient in this series. PMID- 9415939 TI - Adult familial nephrotic syndrome--Balkan variant of congenital nephrotic syndrome. AB - We report on two families from the southern part of Former Yugoslav Republic of Macedonia (close to Mediterranean area) with autosomal dominant inherited nephrotic syndrome, documented with renal biopsy in three family members. Histopathological examination confirmed microcystic tubular changes and focal segmental glomerulosclerosis. In biopsied patients proteinuria was noted in the first decade, or second, but it was quantitatively documented as nephrotic in the second and third decade. The prognosis of the patients was poor: one patient died because of intracranial hemorrhage before developing end stage renal disease; the other two cases developed end stage chronic renal failure in the third and fourth decade. PMID- 9415940 TI - Rapid-onset diabetic nephropathy in type II diabetes mellitus. AB - We report a case of rapidly progressive diabetic nephropathy, from little diabetic change on renal biopsy to severe, nodular, diabetic nephrosclerosis over 32 months. The patient was taking an angiotensin converting enzyme inhibitor and had a mean arterial pressure of 95 mm Hg over this time period. Her dietary protein intake was low, at least upon presentation. She had three additional mechanisms or potential mechanisms of injury: monoclonal kappa light chains; IgA immune deposits on the first, but not the second biopsy; and longstanding hypertension. Her renal histology was typical for diabetic nephropathy but was not characteristic of kappa light chain disease. We suggest that diabetic nephropathy may develop more rapidly than previously assumed, especially when additional mechanisms of injury, or additional promoters of mesangial matrix accumulation are present. PMID- 9415941 TI - Clinical economics and sleep disorders. AB - Sleep disorders have been shown to have substantial psychosocial sequelae with large economic impact. Numerous studies have examined the psychosocial aspects of sleep disorders; however, there has been little published on the associated economic implications. With increasing pressure to contain health care expenditures and provide value for the dollar, clinical economics is playing an important role in the decision-making process about alternative strategies within health care organizations. There are several strategies one may pursue to examine the economics of medical interventions. The predominant strategies include: cost identification, cost effectiveness, cost utility, and cost benefit. This review provides a basis for performing clinical economic evaluations in sleep disorders. PMID- 9415942 TI - An instrument to measure functional status outcomes for disorders of excessive sleepiness. AB - This article reports the development of the functional outcomes of sleep questionnaire (FOSQ). This is the first self-report measure designed to assess the impact of disorders of excessive sleepiness (DOES) on multiple activities of everyday living. Three samples were used in the development and psychometric analyses of the FOSQ: Sample 1 (n = 153) consisted of individuals seeking medical attention for a sleep problem and persons of similar age and gender having no sleep disorder; samples 2 (n = 24) and 3 (n = 51) were composed of patients from two medical centers diagnosed with obstructive sleep apnea (OSA). Factor analysis of the FOSQ yielded five factors: activity level, vigilance, intimacy and sexual relationships, general productivity, and social outcome. Internal reliability was excellent for both the subscales (alpha = 0.86 to alpha = 0.91) and the total scale (alpha = 0.95). Test-retest reliability of the FOSQ yielded coefficients ranging from r = 0.81 to r = 0.90 for the five subscales and r = 0.90 for the total measure. The FOSQ successfully discriminated between normal subjects and those seeking medical attention for a sleep problem (T157 = -5.88, p = 0.0001). This psychometric evaluation of the FOSQ demonstrated parameters acceptable for its application in research and in clinical practice to measure functional status outcomes for persons with DOES. Thus, the FOSQ can be used to determine how disorders of excessive sleepiness affect patients' abilities to conduct normal activities and the extent to which these abilities are improved by effective treatment of DOES. PMID- 9415943 TI - Daytime sleepiness and sleep habits of Australian workers. AB - Excessive daytime sleepiness in the general community is a newly recognized problem about which there is little standardized information. Our aim was to measure the levels of daytime sleepiness and the prevalence of excessive daytime sleepiness in a sample of Australian workers and to relate that to their self reported sleep habits at night and to their age, sex, and obesity. Sixty-five percent of all 507 employees working during the day for a branch of an Australian corporation answered a sleep questionnaire and the Epworth sleepiness scale (ESS) anonymously. Normal sleepers, without any evidence of a sleep disorder, had ESS scores between 0 and 10, with a mean of 4.6 +/- 2.8 (standard deviation). They were clearly separated from the "sleepy" patients suffering from narcolepsy or idiopathic hypersomnia whose ESS scores were in the range 12-24, as described previously. ESS scores > 10 were taken to represent excessive daytime sleepiness, the prevalence of which was 10.9%. This was not related significantly to age (22 59 years), sex, obesity, or the use of hypnotic drugs but was related significantly but weakly to sleep-disordered breathing (frequency of snoring and apneas), the presence of insomnia, and reduced time spent in bed (insufficient sleep). PMID- 9415944 TI - HLA haplotypes, polysomnography, and pedigrees in a case series of patients with narcolepsy. AB - An ongoing study of the genetics of narcolepsy ascertains families through a case series of narcoleptic probands using diagnostic criteria consisting of 1) clinical history of excessive somnolence, 2) a mean sleep latency on the multiple sleep latency test (MSLT) of less than 7.9 minutes, 3) the rapid eye movement (REM) sleep-related symptom of cataplexy, 4) nocturnal polysomnography ruling out sleep apnea syndrome, and 5) two or more transitions to REM sleep on the MSLT. All probands and first-degree relatives received clinical and laboratory evaluations as well as human leukocyte antigen (HLA) typing. Demographic characteristics of the 32 probands are as follows: 17 males and 15 females; mean age was 42.1 years (range 13-70 years). The polysomnographic data confirmed daytime sleepiness and increased tendency for REM sleep for the 32 probands. Nocturnal polysomnographic results are as follows: sleep latency, 3.2 minutes; total sleep time, 442 minutes. MSLT results are as follows: sleep latency, 3.1 minutes; REM latency, 6.9 minutes; number of REM periods, 3.2. HLA typing revealed the presence of the HLA haplotypes, DRB1*15 and DQB1*0602, in 21 narcoleptic probands, with two African-Americans having the DQB1*0602 but not the DRB1*15 allele. Among the 57 relatives of the 32 probands, 1/31 females and 7/26 males were found to be affected with narcolepsy (p < 0.02), which suggests a higher diagnostic rate in male relatives. The 21 probands who were positive for the DRB1*15 and DQB1*0602 haplotypes did not differ from the 10 probands who were negative for these alleles in terms of their nocturnal sleep parameters, MSLT findings, or clinical presentation. Three families with multiple individuals affected with narcolepsy are presented. Two families have more than one affected individual who does not have the high-risk HLA haplotype. In one of these families, the disease is segregating independently of any HLA haplotype. In the third family, there is cosegregation with HLA DRB1*15 and DQB1*0602. One family contains a pair of DNA-confirmed, monozygotic twins with narcolepsy who are discordant for cataplexy and have the HLA DR14(Dw9)/DQB1*0503 and DR4(Dw4)/DQB1*0302 haplotypes. PMID- 9415945 TI - The effects on human sleep and circadian rhythms of 17 days of continuous bedrest in the absence of daylight. AB - As part of a larger bedrest study involving various life science experiments, a study was conducted on the effects of 17 days of continuous bedrest and elimination of daylight on circadian rectal temperature rhythms, mood, alertness, and sleep (objective and diary) in eight healthy middle-aged men. Sleep was timed from 2300 to 0700 hours throughout. Three 72-hour measurement blocks were compared: ambulatory prebedrest, early bedrest (days 5-7), and late bedrest (days 15-17). Temperature rhythms showed reduced amplitude and later phases resulting from the bedrest conditions. This was associated with longer nocturnal sleep onset latencies and poorer subjectively rated sleep but with no reliable changes in any of the other sleep parameters. Daily changes in posture and/or exposure to daylight appear to be important determinants of a properly entrained circadian system. PMID- 9415946 TI - Sleep loss results in an elevation of cortisol levels the next evening. AB - Sleep curtailment constitutes an increasingly common condition in industrialized societies and is thought to affect mood and performance rather than physiological functions. There is no evidence for prolonged or delayed effects of sleep loss on the hypothalamo-pituitary-adrenal (HPA) axis. We evaluated the effects of acute partial or total sleep deprivation on the nighttime and daytime profile of cortisol levels. Plasma cortisol profiles were determined during a 32-hour period (from 1800 hours on day 1 until 0200 hours on day 3) in normal young men submitted to three different protocols: normal sleep schedule (2300-0700 hours), partial sleep deprivation (0400-0800 hours), and total sleep deprivation. Alterations in cortisol levels could only be demonstrated in the evening following the night of sleep deprivation. After normal sleep, plasma cortisol levels over the 1800-2300-hour period were similar on days 1 and 2. After partial and total sleep deprivation, plasma cortisol levels over the 1800-2300-hour period were higher on day 2 than on day 1 (37 and 45% increases, p = 0.03 and 0.003, respectively), and the onset of the quiescent period of cortisol secretion was delayed by at least 1 hour. We conclude that even partial acute sleep loss delays the recovery of the HPA from early morning circadian stimulation and is thus likely to involve an alteration in negative glucocorticoid feedback regulation. Sleep loss could thus affect the resiliency of the stress response and may accelerate the development of metabolic and cognitive consequences of glucocorticoid excess. PMID- 9415947 TI - Sleep deprivation affects speech. AB - Historical accounts of sleep loss studies have described changes in the content and patterns of speech, although to date these claims have not been systematically studied. We examined the effects of sleep loss on the spontaneous generation of words during a verbal word fluency task and the articulation of speech during a vocalized reading task. Nine subjects underwent two counterbalanced 36-hour trials involving sleep deprivation (SD) and no sleep deprivation (NSD). After SD, there was a significant deterioration in word generation and a tendency for subjects to become fixated within a semantic category. There was a significant reduction in the subjects' use of appropriate intonation in the voice after SD, with subjects displaying more monotonic or flattened voices. These findings are discussed in light of neuropsychological evidence concerning the functions of sleep in relation to the frontal cortex and in light of the implications for interpersonal communication in the event of sleep loss. PMID- 9415948 TI - Pergolide in the management of restless legs syndrome: an extended study. AB - Twenty patients with problematic restless legs syndrome (RLS) were treated with pergolide. Efficacy, dosage, side effects, and tolerance were analyzed. Fifteen patients continued treatment for a median study time of 2 years. Five patients discontinued treatment after a mean of 4.2 months. Pergolide resulted in complete or near complete control of symptoms in 45% and moderate control in 50% of patients studied. Levodopa-induced daytime augmentation resolved in all patients in whom it had been present. The mean total daily maintenance dose of pergolide was 0.23 mg. Forty percent required an additional afternoon dose. Side effects developed in 12 patients (60%) and necessitated discontinuation of treatment in five. Common side effects were nausea, dizziness, and insomnia. Daytime augmentation occurred in 27% of patients, but this was mild and usually easily controlled with a supplementary afternoon dose of pergolide. Tolerance did not develop. We conclude that pergolide is an effective second-line agent for RLS, especially following levodopa-induced daytime augmentation. PMID- 9415949 TI - Simultaneous digitization of upper airway fiber-optic images and respiratory signals. AB - We developed an inexpensive and efficient method for simultaneously digitizing respiratory signals and fiber-optic images of the upper airway. The main components of the system are a fiber-optic scope, a charge coupled device video camera, and a personal computer equipped with a frame grabber and an A/D board. The frame grabber digitizes images at five frames per second while the A/D board samples six respiratory signals at 25 samples per second. Digitized images are saved only in the event that the user instructs the computer to do so in order to limit disk space requirements. A circular buffering technique provides continuous storage of the most recent 50 frames in frame grabber memory. This feature gives the user up to 10 seconds, following the beginning of a respiratory event, to initiate the saving of images to computer hard disk. A postacquisition program displays the data acquired during the sleep study and allows the user to interactively select images for subsequent upper airway area measurement. This system enables us to observe and quantify the dynamics of the upper airway during different breathing conditions with minimal time and cost. It is also a potential clinical tool to use to determine the site of obstruction during sleep in patients with obstructive sleep apnea. PMID- 9415950 TI - Subclinical REM sleep behavior disorder in a patient with corticobasal degeneration. AB - Various neurodegenerative diseases have been reported to be associated with rapid eye movement (REM) sleep behavior disorder (RBD). This is the first report of a patient with corticobasal degeneration (CBD) associated with subclinical RBD. A 72-year-old woman was admitted complaining of fine tremor of the right hand and weakness of the right lower extremity. She was diagnosed as having CBD on the basis of clinical features and neuroimaging studies. Her family noticed snoring and increase in sleep talk, but they did not regard them as pathological. All night polysomnography (PSG) revealed REM sleep without atonia (RWA) during which 14 episodes of talking and singing were observed. They ranged from the utterance of one word to that of comprehensible words of a song for about 3 minutes accompanied by various nonpurposeful movements of the mouth, hands, and limbs. These episodes were not associated with any sleep-disturbed breathing. Future PSG studies on CBD patients together with postmortem analysis of brain stem structures that are crucial for generating REM sleep-related atonia are warranted for further understanding of the pathophysiological mechanism of RBD. PMID- 9415951 TI - Unusual complication of nasal CPAP: subcutaneous emphysema following facial trauma. AB - Subcutaneous emphysema is an unusual complication of nasal continuous positive airway pressure (CPAP). We report a case of a 58-year-old man who fell and sustained mild facial trauma to the left side of his head. After using CPAP the following night, he developed diffuse subcutaneous emphysema of his face and left neck. He discontinued CPAP, and his symptoms improved. The potential mechanisms of this patient's subcutaneous emphysema and the prior reports of this complication following facial trauma or dental procedure without use of CPAP are reviewed. Although there are case reports of bacterial meningitis and pneumocephalus following use of nasal CPAP, we are not aware of any prior reports of subcutaneous emphysema following use of CPAP. In light of our experience and the above related case reports, we would suggest nasal CPAP be withheld temporarily in the setting of acute facial trauma. PMID- 9415952 TI - Controversies in sleep medicine: melatonin. PMID- 9415953 TI - Melatonin: a sleep-promoting hormone. AB - This review discusses the issue of a dual effect of melatonin on sleep: acute sleep promotion that typically occurs within one hour of administration, and the ability to alter the phase of an underlying circadian pacemaker after a repeated melatonin treatment. The authors suggest that both mechanisms are at work, that they are complementary, and that they may manifest jointly or separately. The review provides some basic information on melatonin, an overview of the literature, and the authors' experience in studying the acute effects of melatonin treatment in humans of different age groups. This review also illustrates the authors' cautious attitude toward melatonin treatment that induces supraphysiologic circulating levels of the hormone. PMID- 9415954 TI - Sleep-promoting effects of melatonin: at what dose, in whom, under what conditions, and by what mechanisms? AB - Differing conclusions regarding the sleep-promoting effects of melatonin may be the result of the broad range of doses employed (0.1-2000 mg), the differing categories of subjects tested (normal subjects, insomniac patients, elderly, etc.), and the varying times of administration (for daytime vs. nighttime sleep). We conclude that melatonin may benefit sleep by correcting circadian phase abnormalities and/or by a modest direct soporific effect that is most evident following daytime administration to younger subjects. We speculate that these effects are mediated by interactions with specific receptors concentrated in the suprachiasmatic nucleus (SCN) that result in resetting of the circadian pacemaker and/or attenuation of an SCN-dependent circadian alerting process. PMID- 9415955 TI - A critical evaluation of the hypnotic efficacy of melatonin. AB - This paper reviews the available literature on the use of melatonin as a hypnotic in normal volunteers and in patients with noncircadian insomnias. Data are ordered in terms of studies of subject self-reports and polygraphic data and are seen in the context of generally accepted criteria for assessment of clinically used hypnotics. It is concluded that, at the present time, there is very little systematic evidence to suggest that melatonin has significant hypnotic efficacy in these populations. PMID- 9415957 TI - Bibliography of recent literature in sleep research. PMID- 9415956 TI - Do beta-blockers pose an unacceptable risk to patients with obstructive sleep apnea (OSA)? PMID- 9415958 TI - Child sexual abuse accommodation evidence: the travails of counterintuitive evidence in Australia and New Zealand. AB - The author advances a taxonomy of expert evidence in relation to the responses of sexually abused children to their assaults. He analyses a series of Australian and New Zealand cases in the context of Summit's publicly stated recent views on the use to which Child Sexual Abuse Accommodation Syndrome ("CSAAS") can be put. He argues that expert evidence that goes further than dispelling myths or disabusing triers of fact of misperceptions generally held within the community should not be permitted. He contends that CSAAS is an illegitimate description of such information and that such "syndrome evidence" should not be permitted, particularly before juries. He maintains, moreover, that further rigour will be (and should be) required of prosecutors and mental health professionals alike if such counterintuitive evidence is to be admitted. This will entail proof of the existence of community misunderstanding of the phenomenon, establishment that the field of expertise from which child sexual abuse accommodation evidence emanates is sufficient for the purpose, as well as proof of the expert's expertise in the field. However, he argues against abandoning attempts to use counterintuitive evidence merely on the basis of a number of adverse Australasian determinations, whose reasoning is dependent on the quality of the evidence hitherto advanced and the overly limited perceptions of the evidential framework within which such evidence should be evaluated. PMID- 9415959 TI - The globalization of behavioral science evidence about battered women: a theory of production and diffusion. AB - A theoretical framework is proposed for understanding how the innovative use of behavioral science evidence is both produced and diffused among members of the global legal community. Using case law analyses and interviews with key individuals involved in selected cases, we examine how battered woman syndrome (BWS) is produced and diffused between and among Australia, Canada, England, and the United States. The following diffusion mechanisms are proposed: (1) The availability and accessibility of credible dissemination sources; (2) characteristics of the overall practice environment operating in each legal culture; (3) the attitudes and knowledge of attorneys and judges about the use of scientific evidence; (4) political and social support for the use of the evidence in the legal culture; and (5) the level of structural equivalence, communication, and "neighbor effects" between and among legal cultures. Each mechanism is discussed and supplemented with information from interviews with individuals involved in key cases involving BWS evidence. PMID- 9415960 TI - The MacArthur Adjudicative Competence Study: diagnosis, psychopathology, and competence-related abilities. AB - A set of measures assessing abilities related to legal standards for competence in the adjudicative process were administered to mentally-disordered criminal defendants with diagnoses of schizophrenia, affective disorder, other psychiatric disorders, and to criminal defendants without diagnosed mental disorder. Mentally disordered defendants were recruited from two groups: those who had been committed for restoration of competence and those who had been identified by jail personnel as mentally ill. Significant impairments in competence-related abilities were found for approximately half of the defendants with schizophrenia. Defendants with schizophrenia scored lower on measures of understanding, reasoning, and appreciation related to the adjudication process. The association between symptoms and competence-related abilities was explored within diagnostic groups. Conceptual disorganization was found to be inversely correlated with performance on all measures in both defendants with schizophrenia and those with affective disorders. For other psychotic symptoms, differing patterns of correlations were found in the two major diagnostic groups. The implications for policy designed to safeguard the rights of defendants to be tried while competent are discussed. PMID- 9415961 TI - Five year research update (1991-1995): evaluations for competence to stand trial. AB - This article reviews and evaluates publications during 1991-1995 with relevance for assessments of competence to stand trial. The review focuses specifically on articles that provide new concepts or data supported by research or case analyses. The studies are reviewed under the following headings: (a) the systemic context of competence to stand trial (CST) evaluations, (b) conceptual definitions of CST, (c) research on CST assessment methods, (d) characteristics of incompetent defendants, (e) interpretation and communication of CST evaluation data, (f) issues in CST assessment of special populations (juveniles, persons with mental retardation), and (g) treatment to restore competence. Suggestions are offered for further research to advance the quality of clinical evaluations for competence to stand trial. PMID- 9415962 TI - Prolonged use of coloured overlays for classroom reading. AB - Ninety-three children in a primary school and 59 children in two first-year classes of a secondary school were asked individually to observe a paragraph of random letters arranged to resemble text, and to compare the perceptual effects on its clarity of coloured plastic sheets overlaid on the text. A total of 29 colours were compared using 10 coloured plastic sheets and 19 pairwise combinations of sheets, one superimposed on another. The resulting colours sampled CIE 1976 hue angle (huv) and saturation (suv) systematically and efficiently. All the children who reported beneficial perceptual effects (53 per cent) were given their preferred overlay or combination of overlays to use as and when they wished. When the children were examined three months later the children tended to choose a colour similar to one they had chosen previously. Ten months later, 22 per cent of those offered the overlaps were still using them of their own volition. These children, but not those who had ceased to use their overlay(s), read randomly ordered simple words more quickly with their overlay than without. In a second independent group of children referred to the Norfolk Sensory Support Service, who used overlays routinely, the reading speed was similar with a grey or clear overlay; and slower than with the chosen coloured overlay, suggesting that reduction of contrast was not the critical factor. In a third independent group of children in a primary school in Kent, the increase in reading speed with the chosen overlay predicted the children who continued to use their overlay during the ensuing eight weeks. PMID- 9415963 TI - Handedness dependency in recall from everyday memory. AB - A number of previous studies have demonstrated systematic misremembering of the direction in which the Queen's head faces on British coins. Two experiments were carried out to investigate whether this phenomenon is affected by a person's handedness. In both experiments, right-handed and left-handed participants were found to differ significantly in their verbal responses, with recall performance significantly worse than chance for right-handed but not for left-handed participants. Experiment 2 also examined degrees of handedness, and found significant variation in recall across the handedness range. Performance in this everyday-memory paradigm appears to be determined by both handedness and schema factors. It is proposed that although in this task the response was verbal one, relevant motor imagery may nevertheless have been activated and led to the highly unusual observation of an effect of handedness upon cognitive performance. PMID- 9415965 TI - Sex differences on Bartel's task: an investigation into perception of real and depicted distances. AB - A new method derived from Bartel's (1958) studies was used to investigate sex differences in spatial perception. Bartel employed two related tasks; one of these tasks called for responses to a pictorial stimulus representing spatial arrangement in perspective and the other for responses to an analogous task presented in three dimensions. Modified forms of both these tasks were used. Consistent differences between men and women were found, the men showing greater distance constancy in relation both to real and to depicted distances. In addition a decline of such constancy with age was observed. The pictorial task was also used to test two groups of students of architecture: one relatively inexperienced and the other more experienced with the discipline, on the assumption that experience of spatial judgments might influence performance on this task. It was found that whilst familiarity did not affect responses there was a consistent difference between responses of men and women. PMID- 9415964 TI - Testing the hypothesis of the relationships between laterality and ability according to Annett's right-shift theory: findings in an epidemiological sample of young adults. AB - In a large epidemiological sample of young adults, predictions of the right-shift (RS) theory of Annett that cognitive abilities will vary with right-left hand skill were tested. Presenting a theory of a genetic balanced polymorphism with heterozygote advantage for laterality and ability, Annett & Manning (1989, 1990a) and Annett (1993c) claimed that probands at the right end of the R-L hand skill continuum would show lower general intelligence in IQ testing and that specific verbal abilities and educational success would be lower at both extremes of the R L distribution, taking the form of an inverted U. Most of these predictions could not be confirmed by our study. In particular, our data contrast with the important and specific prediction of the RS theory that strong dextrals will be the most disfavoured group. In our sample, probands at the left end of the R-L continuum had significantly lower scores in spelling and educational success and showed a tendency to have lower non-verbal IQ scores, while strong dextrals tended to have average or even marginally higher ability scores. The effects, however, are small and decrease when controlling for other variables. Implications of these empirical findings for the right-shift theory are discussed. PMID- 9415966 TI - Arousal, sensation seeking and frequency of gambling in off-course horse racing bettors. AB - Heart rate was recorded in a sample of 32 off-course horse racing bettors before, during and after the gambling process, together with sensation seeking and information regarding frequency and expenditure on gambling. Importantly, the present study controlled for the confounding effects of movement on heart rate present in previous studies. Significant differences within subjects were found for heart rate at different points in the gambling process, but no differences were found comparing high- and low-frequency gamblers or gamblers that chase with those that don't chase. These findings are discussed in relation to the theoretical role of arousal and sensation seeking in the explanation of gambling behaviour. PMID- 9415967 TI - Diltiazem inhibits fatty acid oxidation in the isolated perfused rat liver. AB - The effects of diltiazem on fatty acid metabolism were measured in the isolated perfused rat liver and in isolated mitochondria. In the perfused rat liver diltiazem inhibited oxygen uptake and ketogenesis from endogenous substrates. Ketogenesis from exogenously supplied palmitate was also inhibited. The beta hydroxybutyrate/acetoacetate ratio in the presence of palmitate alone was equal to 3.2. When the fatty acid and diltiazem were present simultaneously this ratio was decreased to 0.93, suggesting that, in spite of the inhibition of oxygen uptake, the respiratory chain was not rate limiting for the oxidation of the reducing equivalents coming from beta-oxidation. In experiments with isolated mitochondria, incubated in the presence of all intermediates of the Krebs cycle, pyruvate or glutamate, no significant inhibition of oxygen uptake by diltiazem was detected. Inhibition of oxygen uptake in isolated mitochondria was found only when palmitoyl CoA was the source of the reducing equivalents. It was concluded that a direct effect on beta-oxidation may be a major cause for the inhibition of oxygen uptake caused by diltiazem in the perfused liver. PMID- 9415968 TI - Polyamine metabolism in various tissues during pathogenesis of chloroquine susceptible and resistant malaria. AB - The pathophysiological impact of infections with chloroquine-susceptible (CQS) and chloroquine-resistant (CQR) strains of Plasmodium berghei in Mastomys natalensis was studied with respect to changes in polyamine profiles in various tissues. Both CQS and CQR infections produced similar changes in polyamine profiles of various tissues. Maximum increase was recorded in spleen followed by liver and lungs. Renal, cardiac and cerebral tissues did not register significant changes. An increase in spermidine level was more prominent as compared to putrescine and spermine, leading to an overall increase in spermidine/spermine ratio. This ratio is an important index of cellular proliferation. Liver did not show considerable change in the activities of ornithine decarboxylase and S adenosyl methionine decarboxylase, the regulatory enzymes of the polyamine biosynthetic pathway. Spleen however, registered marked induction of both the enzymes which was more prominent in the CQS infection than CQR. Normal erythrocytes contained traces of polyamine while the erythrocytes loaded with P. berghei parasites exhibited appreciably higher polyamine levels. Spermidine was detected in about five-fold higher concentrations than putrescine and spermine which were detected in equimolar levels. Again, CQS as well as CQR P. berghei, exhibited qualitatively and quantitatively similar polyamine profiles thus ruling out a role of polyamines in CQ-resistance in malaria. PMID- 9415969 TI - Non-functional role of syntaxin 2 in insulin exocytosis by pancreatic beta cells. AB - This study was designed in order to examine the expression and functional role of syntaxin 2/epimorphin in pancreatic beta cells. Northern blot analysis revealed that syntaxin 2 mRNA was able to be detected in mouse beta TC3 cells, but not in isolated mouse islets. In agreement with this result, immunoblot analysis detected an appreciable amount of syntaxin 2 protein in beta TC3 cells, but not in mouse islets. Immunohistochemistry of the mouse pancreas demonstrated that syntaxin 2 was little evident in islet cells of Langerhans, and somewhat predominant in exocrine tissues. In order to examine whether syntaxin 2 is anchored to cell surfaces in beta TC3 cells, living cells were incubated with a monoclonal antibody against syntaxin 2 (MC-1). The antibody bound to their surfaces, indicating that syntaxin 2 was localized on cell surfaces. The addition of MC-1 to the culture medium of beta TC3 cells did not affect insulin release under the presence or absence of 11 mM glucose, indicating that syntaxin 2 is not associated with insulin exocytosis. Thus, the expression of syntaxin 2 in islets of Langerhans is very low and the function of this protein is probably unrelated to the insulin exocytosis pathway. PMID- 9415970 TI - Novel monosaccharides as potent inhibitors of cell proliferation. AB - The effects of several novel monosaccharides upon thymidine incorporation into both normal and tumour cells were investigated. The monosaccharide 2-deoxy-3-[1 (R)-(ethoxycarbonyl)ethyl]- alpha-D-allo-pyranose had the most inhibitory effect on proliferation, with the (S)-enantiomer having less inhibitory effects. The chiral centre at carbon-7 was found to be an important part of the molecule, as 2 deoxy-3-[methoxycarbonyl methyl]-alpha-D-allo-pyranose had greatly decreased anti proliferative properties in comparison with the parent compound. In addition, the 2-deoxy structure at carbon-2 was also found to be important, as 3-[1-(S) (ethoxycarbonyl)ethyl]-alpha-D-allo-hexopyranose had greatly decreased inhibitory properties in comparison with the parent compound. The results indicate that these novel monosaccharides possess potent anti-proliferative properties, related to their chiral carbon-7 and 2-deoxy carbon-2 structure and suggest that further substitutions of the functional group at carbon-7 may improve these properties and possibly produce inhibitor selectivity for tumour cells in preference to normal cells. PMID- 9415971 TI - Effect of heat shock treatment on the production of variant testosterone repressed prostate message-2 (TRPM-2) mRNA in culture cells. AB - The testosterone-repressive prostate message-2 (TRPM-2) variant mRNA lacking the exon 5 was induced in rat primary culture hepatocytes by heat shock treatment. A similar variant mRNA lacking exon 5 was also induced by heat shock treatment of the human culture cell line HepG2. On the other hand, in mouse cell line L929, heat shock treatment induced a variant TRPM-2 mRNA lacking only a small region located in exon 5. However, irrespective of the difference of mechanism of variant production, all the variant TRPM-2 mRNA species derived from each animal species encoded a putative protein constituted from the N-terminal one-third of TRPM-2 protein attached to a C-terminal TRPM-2 unrelated tail. In humans, the variant TRPM-2 species was not detected in normal tissues but was present in certain kinds of tumour cells. These results indicate that the splicing variants were induced as a direct result of heat shock treatment on cells per se and that the phenomenon of heat shock induction was observed in culture cells derived from different animal species. PMID- 9415972 TI - Effect of methotrexate (MTX) on NAD(P)+ dehydrogenases of HeLa cells: malic enzyme, 2-oxoglutarate and isocitrate dehydrogenases. AB - The effects of methotrexate (MTX) on oxygen uptake by permeabilized HeLa cells were evaluated. MTX did not inhibit state III respiration when the oxidizable substrate was succinate, but when the substrates were 2-oxoglutarate or isocitrate the respiration decreased about 50 per cent at 1.0 mM concentration of the drug. This effect was explained by inhibition of 2-oxoglutarate and isocitrate dehydrogenases by MTX. No effect was observed on succinate dehydrogenase. An evaluation of the effects of MTX on malic enzyme activity as measured by pyruvate plus lactate production in intact cells supplied with malate showed a decrease of about 40 per cent in metabolite production using 0.4 mM MTX. HeLa cell malic enzyme, as observed for other tumour cells, is compartmentalized in mitochondria and cytosol, and is another example of a dehydrogenase inhibited by MTX. PMID- 9415973 TI - The effects of selenium, vitamin E and their combination on the composition of fatty acids and proteins in Saccharomyces cerevisiae. AB - The aim of our studies was to test the effect and role of vitamin E and selenium supplements on yeast cell. In this study, the effects of selenium (Se), vitamin E (Vit. E), and their combination (Se plus Vit. E) on the composition of fatty acids and proteins were examined in Saccharomyces cerevisiae strains WET136 and 522. S. cerevisiae cells were grown up in YEPD medium supplemented with Se, Vit. E or their combination. It was found that the level of stearic acid was increased in all supplemented groups (p < 0.05; p < 0.001). The content of saturated and unsaturated fatty acids was decreased (p < 0.05; p < 0.01; p < 0.001) in Vit. E and Vit. E plus Se supplemented S. cerevisiae. On the other hand, Se alone caused an increase (p < 0.001) in the saturated fatty acids but a decrease (p < 0.05; p < 0.001) in the unsaturated fatty acids. Total proteins in S. cerevisiae were significantly increased (p < 0.001) by Vit. E supplement. There was no significant change observed in S. cerevisiae supplemented with Se. These findings indicate that membrane composition of S. cerevisiae is affected by both Vit. E and Se supplements. PMID- 9415974 TI - Antagonism by ethanol of endotoxin-induced tissue factor activation in relation to the depressed endotoxin binding to monocyte-like U937 cells. AB - Our previous study has reported that ethanol (ETOH) partially inhibited the endotoxin (LPS)-induced tissue factor (TF)-activation in monocytes including blood peripheral monocytes as well as cultured leukemic U937 and THP-1 cells. The present study shows a strong correlation (r = 0.92; p < 0.01) between TF activation and depression in LPS binding blocked by ETOH in U937 cells. The antagonism by ETOH of LPS binding was not due to a direct extracellular blockade, since ETOH did not affect the affinity of fluorescein isothiocyanate (FITC)-LPS or -anti CD14 mAb on U937 cells. After U937 cells were treated with 2 per cent (v/v) ETOH for 3 h, LPS binding was however drastically inhibited as shown by immunostaining with FITC-LPS which was viewed on a confocal laser scanning microscope. The results imply that cellular events of the ETOH effect mediate this inhibition of LPS binding. Anti-CD14 mAb (UCHM-1) inhibited LPS binding in a dose-dependent fashion, revealing a competitive specific binding to the LPS receptor. The results suggest that CD14 plays an important role in the recognition of LPS. FITC-UCHM-1 binding was significantly reduced in the cells pretreated with 2 per cent (v/v) ETOH for 3 h, indicating that ETOH modulates the ability to express CD14. CD14 expression was upregulated by priming with LPS which was offset by ETOH. Acetaldehyde, a possible metabolite of ETOH, was tested with no effect on CD14 expression. Taken together, our results show that ETOH downregulates the recognition of LPS, and suggest that the inhibitory action is likely to be mediated by the depression in CD14 expression which was also accompanied by a significantly altered membrane fluidity. Thus, the antagonism by ETOH of the binding of LPS results in a depression in the LPS-induced TF activation. PMID- 9415975 TI - Osteogenesis imperfecta mutations may probe vital functional domains (e.g. proteoglycan binding sites) of type 1 collagen fibrils. AB - Osteogenesis imperfecta (OI) is a disease characterized by bone malformations caused by mutations in type 1 collagen. Since many of the 338 possible glycine mutations have not been observed in clinical practice, is this due to chance alone? Because only 83 mutations have been reported in 126 patients, we conclude that many mutations are absent from clinical data for non-random causes. Mutations affecting vital intermolecular interactions in the extracellular matrix (e.g. potential collagen binding sites for proteoglycans) may result in non viable fetuses that do not progress to clinical status. Some mutations may be silent because they do not significantly affect normal function. The total number of clinically active mutations that will be observed may be far fewer than the potential 338 maximum. PMID- 9415976 TI - Effect of dehydroepiandrosterone in hereditarily diabetic rats. AB - Goto-Kakizaki rats (GK rats) were given access for 4 weeks to a diet enriched with dehydroepiandrosterone (DHEA, 0.2 per cent, w/w). The incorporation of DHEA in the food failed to affect significantly body growth, plasma D-glucose and insulin concentrations, pancreatic islet insulin content or the activity of both mitochondrial glycerophosphate dehydrogenase (mGDH) and NADP-malate dehydrogenase (malic enzyme) in islet homogenates. DHEA however, increased the activity of mGDH and, at least in male rates, that of the malic enzyme also in the liver. It lowered the abnormally high basal insulin release otherwise found in the islets from diabetic rats, and, as judged from the ratio of insulin output at 16.7 mM/2.8 mM D-glucose, improved the cell responsiveness to the hexose. This coincided with a decreased plasma insulin/D-glucose ratio, suggesting that the major effect of DHEA was to increase the sensitivity to insulin of extrapancreatic targets, thus resulting in a secondary improvement of cell secretory behaviour. PMID- 9415977 TI - Gonadectomy impairs lymphocyte proliferation and macrophage function in male and female rats. Correlation with key enzyme activities of glucose and glutamine metabolism. AB - The effect of gonadectomy on lymphocyte proliferation and macrophage function (hydrogen peroxide production and phagocytosis capacity) of male and female rats was examined and the results correlated with the activities of hexokinase, glucose-6-phosphate dehydrogenase, citrate synthase and phosphate-dependent glutaminase. Also, the reversion of the changes by the treatment with oestrogen or progesterone or a combination of both was addressed. Taken as a whole, ovariectomy reduced hydrogen peroxide production and phagocytosis capacity by macrophages and also lymphocyte proliferation. Castration of male rats reduced the proliferation of lymphocytes and raised macrophage phagocytosis capacity. The effects on macrophage function were correlated with changes in glucose metabolism, particularly, in the activity of glucose-6-phosphate dehydrogenase. PMID- 9415978 TI - Biodegradation of benzene, toluene, and other aromatic compounds by Pseudmonas sp. D8. AB - Pseudomonas sp. D8 strain, which has the potential to utilize toluene as a sole carbon source, was isolated. At a concentration of 100 mg/l, this strain was found to efficiently degrade toluene and benzene (both individually and in mixture) in culture medium at 30 degrees C and pH7. Following a two-hour lag phase, complete biodegradation of 100 mg/l toluene or benzene occurred within 6 to 8 hours. The addition of nitrate, phosphate, or sulfate at various concentrations were found to have significant influence on both toluene and benzene degradation. In addition, results show that the D8 strain has the ability to degrade monochlorophenols, nitrophenols, and phenol, but not aliphatic compounds. Inoculation of groundwater samples containing 100 mg/l toluene or benzene with Pseudmonas sp. D8 resulted in rapid degradation within 24-33 hours. PMID- 9415979 TI - Tissue distribution of inorganic mercury, methylmercury and cadmium in the Asiatic clam (Corbicula fluminea) in relation to the contamination levels of the water column and sediment. AB - The comparative experimental study of inorganic mercury (HgII), methylmercury (MeHg) and cadmium (Cd) bioaccumulation in the Asiatic clam Corbicula fluminea was based on a 14 days' exposure to the water column or sediment compartments, as initial contamination sources. For each contaminant and exposure source, a five point concentration range was set up in order to quantify the relationships between the contamination pressure and bioaccumulation capacity, at the whole soft body level and in five organs: gills, mantle, visceral mass, kidney and foot. Hg and Cd bioaccumulation at the whole organism level was proportional to the metal concentrations in the water column or sediment. For similar exposure conditions, the average ratios between the metal concentrations in the bivalves- [MeHg]/[HgII] and [MeHg]/[Cd]--were close to 10 and 5 for the sediment source and 8 and 15 for the water column source. Metal distribution in the five organs revealed strong specificities, according to the different contamination modalities studied: kidney and gills were clearly associated with Cd exposure, mantle and foot with MeHg exposure and the visceral mass with inorganic Hg exposure. PMID- 9415980 TI - Total mercury intake from fish and shellfish by Japanese people. AB - Elevated mercury concentrations have been reported in fish in recent years. Japanese people eat a great deal of raw fishes and shellfishes as "Sashimi" and "Sushi". The action level of large predatory fish such as tuna with total mercury levels exceeding the Japanese maximum permitted limit of 0.4 ppm is exempted from regulation in Japan. Therefore, current total mercury intake from fish and shellfish of Japanese people is unknown. The purpose of this investigation was to estimate the total mercury intake from fish and shellfish. It was found that the mean total mercury concentration of 1.11 ppm in tuna of eatable base as Sashimi or Sushi was clearly higher than the normal level. The mean total mercury intake from fish and shellfish was 0.17 mg per capita per week. According to the hypothesis that 75% of total mercury in fish and shellfish is methylmercury, the weekly intake of 0.13 mg as methylmercury was corresponding amount to about 74% of provisional tolerable weekly intake 0.17 mg of methylmercury set by the Welfare Ministry of Japan. PMID- 9415981 TI - Testing biodegradability with standardized methods. AB - Laboratory test methods are used by industry laboratories to determine biodegradability, an important parameter for the evaluation of the ecological behaviour of substances. Biodegradability has a key role due to the simple fact that a degradable substance will cause no long term risk in the environment. The great variety of biodegradation processes in the natural environment and in technical plants for treating waste water and solid wastes gave rise to a rather large number of test methods based on different test principles. To guarantee the acceptance of the test results by authorities and customers internationally standardized methods (ISO, OECD) and established quality criteria (GLP, EN 45,000, ISO 9000) are used. PMID- 9415982 TI - Environmental risk assessment of musk ketone and musk xylene in The Netherlands in accordance with the EU-TGD. AB - An environmental risk assessment has been carried out for musk ketone and musk xylene according to the EU Technical Guidance Document for Environmental Risk Assessment for New and Existing Substances [1]. Musk ketone and musk xylene are used in fragrances for cosmetics and household products. For the fragrance industry these are important fragrance ingredients because of their excellent substantivity as well as for their unique smell, which determines largely the odor of a product. The initial environmental risk assessment is based on information provided by the fragrance industry as represented in the Netherlands by its association NEA, by the Research Institute for Fragrance Materials (RIFM) and data reported in the international open literature. The risk assessment includes and evaluation of the risks for aquatic organisms in surface water and sediment and for soil organisms in soil after application of sewage sludge. Secondary poisoning of fish-eating birds and mammals is considered as well. For each compartment the Predicted Environmental Concentration (PEC) is compared to the Predicted No Effect Concentration (PNEC) to obtain PEC/PNEC ratios. Since monitoring data are available in water, sediment and fish, similar ratios are obtained with measured concentrations instead of the predicted ones. For both substances, PEC/PNEC ratios are at or below 0.1 for organisms in the aquatic environment, including sediment organisms. PEC/PNEC ratios for fish-eating predators are 0.01. Ratios based on monitoring data are below 0.01 for all of these organisms. For soil organisms the PEC/PNEC ratio is 0.5 for musk ketone and 1.3 for musk xylene. Although in the Netherlands (as well as in some other European countries), sewage sludge presently finds no application as fertilizer on agricultural soil, the aim of environmental policy is to upgrade the sludge quality to enable future applications on agricultural and grassland. The reliability of the predicted soil concentrations can be greatly improved by obtaining experimental data on fate and behaviour of musk ketone and musk xylene in digested sludge and soil. The risk assessment provides reassurance for the aquatic compartment while pointing the way for obtaining additional data for the soil compartment. PMID- 9415983 TI - Pesticides in rain. AB - 40 rainwater samples were collected in Hannover and near Peine (Lower Saxony, Germany) in 1992 using a wet-only collector. The samples were extracted by solid phase extraction and analyzed by GC/MS for 59 pesticides. 11 pesticides were found in more than 10 samples. The highest concentrations were observed for terbuthylazine (0.003-0.52 microgram/L), metolachlor (0.003-0.51 microgram/L, mean: 0.10 microgram/L), metalaxyl (0.006-0.48 microgram/L, mean: 0.10 microgram/L) and chlorothalonil (0.003-1.1 micrograms/L, mean: 0.16 microgram/L). The concentrations show a seasonal dependence reflecting the application periods. PMID- 9415984 TI - Evaluating the effectiveness of sequence analysis algorithms using measures of relevant information. AB - Given vast quantities of molecular sequence data, and numerous different algorithms designed to discover, diagnose or model biologically interesting features in sequences, how is it possible to make objective evaluations of the diagnostic effectiveness of these algorithms and robust assessments of their relative strengths and limitations? An approach to this relatively neglected question is developed here, which is based on information measures of the diagnostic efficiency of different methods. From output lists of a procedure such as a database search, "relevance weights" are assigned that encode, for each sequence listed, the level of associated scientific evidence implicating that sequence as an example of a feature of interest. Relevance weights may be derived, following systematic protocols, from expert human judgement or, in principle, by automated information retrieval from electronic resources. Practical applications of this approach to algorithm assessment and development and parameter choice are demonstrated with examples of automated sequence motif modeling for the DNA-binding helix-turn-helix motif and the guanine exchange factor protein domain. The combined use of relevance weights and information measures appears to offer promising advantages over ROC analysis and may be generally applicable to diagnostic evaluation. PMID- 9415985 TI - The difficulty of identifying genes in anonymous vertebrate sequences. AB - The identification of genes in newly determined vertebrate genomic sequences can range from a trivial to an impossible task. In a statistical preamble, we show how "insignificant" are the individual features on which gene identification can be rigorously based: promoter signals, splice sites, open reading frames, etc. The practical identification of genes is thus ultimately a tributary of their resemblance to those already present in sequence databases, or incorporated into training sets. The inherent conservatism of the currently popular methods (database similarity search, GRAIL) will greatly limit our capacity for making unexpected biological discoveries from increasingly abundant genomic data. Beyond a very limited subset of trivial cases, the automated interpretation (i.e. without experimental validation) of genomic data, is still a myth. On the other hand, characterizing the 60,000 to 100,000 genes thought to be hidden in the human genome by the mean of individual experiments is not feasible. Thus, it appears that our only hope of turning genome data into genome information must rely on drastic progresses in the way we identify and analyse genes in silico. PMID- 9415986 TI - Computational gene identification: an open problem. AB - As the Human Genome Project enters the large-scale sequencing phase, computational gene identification methods are becoming essential for the automatic analysis and annotation of large uncharacterized genomic sequences. Currently available computer programs relying mainly on sequence coding statistics are of great use in pin-pointing regions in genomic sequences containing exons. Such programs perform rather poorly, however, when the problem is to fully elucidate gene structure. For this problem, the DNA sequence signals involved in the specification of the genes--start sites and splice sites--carry a lot of information, and simple methods relying on such information can predict gene structure with an accuracy to some extent comparable to that of other more sophisticated computational methods. PMID- 9415987 TI - Artificial neural networks for molecular sequence analysis. AB - Artificial neural networks provide a unique computing architecture whose potential has attracted interest from researchers across different disciplines. As a technique for computational analysis, neural network technology is very well suited for the analysis of molecular sequence data. It has been applied successfully to a variety of problems, ranging from gene identification, to protein structure prediction and sequence classification. This article provides an overview of major neural network paradigms, discusses design issues, and reviews current applications in DNA/RNA and protein sequence analysis. PMID- 9415988 TI - The study of correlation structures of DNA sequences: a critical review. AB - The study of correlation structure in the primary sequences of DNA is reviewed. The issues reviewed include: symmetries among 16 base-base correlation functions; accurate estimation of correlation measures; the relationship between 1/f and Lorentzian spectra; heterogeneity in DNA sequences; different modeling strategies of the correlation structure of DNA sequences; the difference of correlation structure between coding and non-coding regions (besides the period-3 pattern); and source of broad distribution of domain sizes. Although some of the results remain controversial, a body of work on this topic constitutes a good starting point for future studies. PMID- 9415989 TI - Tactic components in orientation. AB - In the first half of this century, taxes were considered the best models for working out the rules of stimulus-response systems. The interest for tactic behaviours suddenly disappeared in the mid-1960s, out of reasons specified in the present review. However, results of several recent studies reviewed in the present article suggest that tactic behaviours constitute, from an ontogenetic as well as phylogenetic point of view, a first step towards more complex oriented behaviours that have received much attention in recent years. The aim of this chapter is to update the implications of tactic responses in complex oriented behaviours. We argue that taxes are basic in the process of acquiring most, if not all of these behaviours, and that they often constitute the first steps in the ontogeny of orientation. Taxes are determined by a flexible balance between genetic and epigenetic factors. Their main function is to assist the ecological adaptation of the animal to the constraints of its environment. Finally, we plead for a revival of the studies of taxes in the light of a theory on the development of behaviour, based upon self-organization of autonomous living systems. PMID- 9415990 TI - Course control and tracking: orientation through image stabilization. AB - Course control and tracking are based on visual detection of the position and movement of objects. A disadvantage of biological movement detectors is that they cannot provide a signal proportional to the speed at which the image of an object moves over the retina. Other image parameters, such as brightness, contrast, and texture, strongly affect the magnitude of the detectors' output signals. To function well, the optomotor control circuit must solve these problems. One possible solution, realized in Diptera, is the principle of "gain control by feedback oscillations" described in this chapter. The optomotor system serves for course control by stabilizing the image of the visual panorama on the eye, and for tracking a moving object by stabilizing the object's image on the eye. When an object moves in front of a structured background, it is impossible for the images of both object and background to be stabilized simultaneously. Arthropods and vertebrates usually employ the same strategy to cope with this problem: saccadic tracking. In Diptera, the neural substrate for saccadic tracking is partially understood. PMID- 9415991 TI - Visual control of honeybee flight. AB - Recent research has uncovered a number of different visual cues which bees use for controlling and stabilising flight. Bees flying through a tunnel maintain equidistance to the flanking walls by balancing the speeds of the images of the two walls. This strategy enables them to negotiate narrow passages or to fly between obstacles. The speed of flight in the tunnel is controlled by holding constant the average image velocity as seen by the two eyes. This mechanism prevents potential collisions by ensuring that the bee slows down when it flies through a narrow passage. Bees landing on a horizontal surface hold constant the image velocity of the surface as they approach it, thus automatically ensuring that flight speed is close to zero at touchdown. The movement-sensitive mechanisms underlying these various behaviours differ qualitatively as well as quantitatively, from those that mediate the well-investigated optomotor response. Flight thus appears to be co-ordinated by a number of visuomotor systems acting in concert. PMID- 9415992 TI - Magnetic orientation and the magnetic sense in arthropods. AB - The physical properties of the earth's magnetic field are summarized with the aim of emphasizing their significance as cues that can be exploited in orientational tasks. Past work has revealed magnetic orientation in vertebrates as well as invertebrates, including arthropods. The key finding to date has been that, as opposed to many vertebrates, the magnetic compass of arthropods responds to the polarity, rather than to the inclination of the earth's magnetic field. As in the case of vertebrates, the debate over how arthropods detect magnetic fields has yet to be resolved. Currently, evidence has been reported in support of a detection system based on magnetite crystals together with a variety of detection systems based on events occurring at the molecular level. Interactions between the magnetic and other compasses in orientation experiments suggest the existence of an area in the brain where spatial orientation information from magnetic and other stimuli converges. The slow advance of our knowledge on magnetic orientation in arthropods, as opposed to the much better understanding of magnetic orientation in vertebrates, arises from difficulties in identifying the appropriate behavioural contexts in which arthropods respond to magnetic fields in both laboratory and field situations. Arthropods thus present challenges not only in demonstrating magnetic orientation, but also in elucidating the sensory mechanisms involved in the perception of magnetic fields. PMID- 9415993 TI - Chemo- and mechanosensory orientation by crustaceans in laminar and turbulent flows: from odor trails to vortex streets. AB - Crustaceans use odor and fluid mechanical cues to extract information from their environment. These cues enable animals to find resources, orient to water currents, or escape predators. Because the properties of the fluid environment affect the transmission and structure of relevant signals, a better understanding of sensory and behavioral mechanisms will be aided by considering, at the same time, the hydrodynamic context of chemo- and mechanosensory behaviors. Crustaceans occupy aquatic habitats where flows range from almost completely laminar to nearly fully turbulent. The considerable scope of hydrodynamic properties is mirrored by equally extreme variations in the complexity of the signals entrained in these flows. Ambient noise and stochastic variation increase in increasingly energetic, turbulent conditions. The sensory and behavioral mechanisms of animals that orient in turbulent environments suggest that they have, in the course of evolution, been shaped by the flow properties. Here, sensory systems are geared to extract rapidly fluctuating signals against a noisy background. They sometimes have elaborate noise filtering mechanisms that enable the detection of rather coarse types of signal features to improve the signal-to noise ratio. In contrast, the simpler and more predictable structure of signals carried in laminar flows may allow more accurate orientation and discrimination to occur, and free animals from the burden of supporting complex noise-filtering circuitry. Future comparative investigations of sensory physiology and behavior of animals in relation to their flow environment promise to increase our understanding of orientation by means of chemo- and mechanoperception. PMID- 9415994 TI - Threshold parameters for a simple stochastic partnership model of sexually transmitted diseases formulated as a two-type CMJ process. AB - A simple stochastic model describing an epidemic of a sexually transmitted disease, accommodating the formation and dissolution of partnerships, was formulated within a two-type Crump-Mode-Jagers (CMJ) process in continuous time. A submodel, describing the formation and dissolution of partnerships, was formulated in terms of a semi-Markov process with a finite state space and death as an absorbing state. This model was then linked to a two-type CMJ process through offspring distributions. Numerical values of threshold parameters greater than one suggests that either the epidemic dies out with a probability significantly different form zero or it spreads explosively in the population. PMID- 9415995 TI - A mathematical model for the capillary endothelial cell-extracellular matrix interactions in wound-healing angiogenesis. AB - Angiogenesis, the process by which new blood capillaries grow into a tissue from surrounding parent vessels, is a key event in dermal wound healing, malignant tumour growth, and other pathologic conditions. In wound healing, new capillaries deliver vital metabolites such as amino acids and oxygen to the cells in the wound which are involved in a complex sequence of repair processes. The key cellular constituents of these new capillaries are endothelial cells: their interactions with soluble biochemical and insoluble extracellular matrix (ECM) proteins have been well documented recently, although the biological mechanisms underlying wound-healing angiogenesis are incompletely understood. Considerable recent research, including some continuum mathematical models, have focused on the interactions between endothelial cells and soluble regulators (such as growth factors). In this work, a similar modelling framework is used to investigate the roles of the insoluble ECM substrate, of which collagen is the predominant macromolecular protein. Our model consists of a partial differential equation for the endothelial-cell density (as a function of position and time) coupled to an ordinary differential equation for the ECM density. The ECM is assumed to regulate cell movement (both random and directed) and proliferation, whereas the cells synthesize and degrade the ECM. Analysis and numerical solutions of these equations highlights the roles of these processes in wound-healing angiogenesis. A nonstandard approximation analysis yields insight into the travelling-wave structure of the system. The model is extended to two spatial dimensions (parallel and perpendicular to the plane of the skin), for which numerical simulations are presented. The model predicts that ECM-mediated random motility and cell proliferation are key processes which drive angiogenesis and that the details of the functional dependence of these processes on the ECM density, together with the rate of ECM remodelling, determine the qualitative nature of the angiogenic response. These predictions are experimentally testable, and they may lead towards a greater understanding of the biological mechanisms involved in wound-healing angiogenesis. PMID- 9415996 TI - The optimal scheduling of two drugs with simple resistance for a problem in cancer chemotherapy. AB - In this paper we study the effects of drug resistance on the optimal schedule of two drugs in cancer chemotherapy. We analytically determine the optimal schedule when the tumour grows exponentially and the drugs have a linear killing action. Drug resistance enters into the model via an effectiveness term that decreases the cell loss as the accumulation of the drug grows. PMID- 9415997 TI - The importance of intercellular adhesion in the development of carcinomas. AB - When grading and classifying tumours several criteria are taken into consideration. These include the type of cell from which the tumour has arisen, whether it is benign or malignant and its ability to invade the surrounding tissue. In this paper we suggest that the ability with which a tumour invades its host environment can be related to the intercellular adhesion forces which maintain the tumour's structure. We develop a mathematical model to describe the growth of an avascular tumour in response to an externally supplied nutrient. Its development depends on the balance between expansive forces caused by cell proliferation and intercellular adhesion forces which maintain the tumour's compactness. We focus attention on the existence, uniqueness, and stability of steady, radially symmetric solutions to the model. Our analysis shows that as the importance of cell-cell adhesion increases the size of the radially symmetric steady tumour radius diminishes and the number of asymmetric modes to which it is stable increases. Thus we conclude that cell-cell adhesion may provide clinicians with a useful index of the invasive potential of a tumour. PMID- 9415998 TI - The mitochondrial processing peptidase. AB - The mitochondrial processing peptidase (MPP) is a heterodimeric enzyme which plays an essential role in mitochondrial protein import. It cleaves off the N terminal targeting signals of nuclear encoded mitochondrial proteins upon their transport into the organelle. In mammals and yeast the enzyme is localized in the mitochondrial matrix while in plants it is integrated into a protein complex of the respiratory chain. As the activity of MPP is essential for the viability of eukaryotic cells it is conceivable that inhibitors of MPP which are specific for the soluble enzyme only present in fungi and animals may work as fungicides or insecticides. PMID- 9415999 TI - Caldesmon. AB - Caldesmon is a protein that is found in smooth muscle and in non-muscle cells. Two isoform classes produced by alternative splicing of one gene have been characterized. The smooth muscle, high molecular weight (89-93 kDa), caldesmon isoforms are exclusively found in adult and fully differentiated smooth muscle cells. The non-muscle, low molecular weight (59-63 kDa), caldesmon isoforms are found in non-muscle and in de-differentiated smooth muscle cells. The conserved regions of all isoforms contain caldesmon's properties such as binding to actin, tropomyosin, Ca(2+)-calmodulin, myosin and phospholipids. All isoforms are also very potent inhibitors of the actin-tropomyosin activated myosin MgATPase. Non muscle and smooth muscle isoforms of caldesmon perform different roles in vivo. This may be reflected by the distinct cellular distribution of these isoform classes. Non-muscle caldesmon is a regulatory factor in the microfilament network and is thus involved in the assembly and stabilization of microfilaments. Smooth muscle caldesmon together with tropomyosin is a mediating factor for Ca(2+) dependent inhibition of smooth muscle contraction. PMID- 9416000 TI - The T cell surface protein, CD28. AB - The cluster of differentiation (CD) antigen CD28 is a 44-kDa, disulphide-bonded, homodimeric glycoprotein, which is constitutively expressed on the surface of all murine T cells and the majority of human T cells. Ligation of CD28 by its counter receptor, B7, expressed on the surface of antigen presenting cells, has been shown to induce signals that, in synergy with those derived from engagement of the T cell receptor by an antigen bound to a major histocompatibility complex, enhance proliferation and cytokine production. Manipulation of this interaction can have dramatic effects on the outcome of T cell activation. Blocking CD28/B7 interactions may be useful in preventing unwanted activation in allergy and autoimmune diseases, whereas enhancing this interaction can promote tumour rejection. Thus, CD28 and its signalling pathways may prove to be useful targets in the development of new therapeutic treatments. PMID- 9416001 TI - Interleukin-11. AB - Interleukin-11 (IL-11) is an IL-6-type cytokine that is produced by a variety of stromal cells including fibroblasts, epithelial cells and osteoblasts. It binds to a multimeric receptor complex which contains an IL-11-specific alpha subunit and a promiscuous 130 kDa beta subunit (gp130). IL-11 stimulates multiple aspects of hematopoiesis and hepatocyte production of acute phase response proteins. It also inhibits the genesis of adipocytes, activates osteoclasts, alters neural phenotype, stimulates tissue fibrosis and regulates chondrocyte, synoviocyte and B cell function. In other settings, IL-11 minimizes tissue injury. This may be the result of its ability to protect clonogenic stem cells, regulate epithelial cell proliferation, inhibit apoptosis and inhibit macrophage cytokine production. Thus, IL-11 appears to play an important role in hematopoiesis, bone metabolism and tissue remodeling and may be an important protector of mucosal surfaces. PMID- 9416002 TI - Scanning transmission electron microscopy mass analysis of fibrillin-containing microfibrils from foetal elastic tissues. AB - We have applied scanning transmission electron microscopy to intact native fibrillin-containing microfibrils isolated from foetal bovine elastic tissues in order to derive new insights into microfibril organisation. This technique provides quantitative data on the mass per unit length and axial mass distribution of unstained, unshadowed macromolecules. Scanning transmission electron microscopy of microfibrils from aorta, skin and nuchal ligament revealed that the beads corresponded to peaks of mass and the interbead regions to troughs of mass. These major features of axial mass distribution were characteristic of all microfibrils examined. Tissue-specific and age-dependent variations in mass were identified in microfibrils that were structurally comparable by rotary shadowing electron microscopy. Increased microfibril mass correlated with increasing gestational age. The additional mass was associated predominantly at, or close to, the bead. Some microfibril populations exhibited pronounced assymetry in their axial mass distribution. These data indicate that intact native microfibrillar assemblies from developing elastic tissues are heterogeneous in composition. Loss of mass following chondroitinase ABC or AC lyase treatment confirmed the presence of chondroitin sulphate in nuchal ligament microfibrillar assemblies. PMID- 9416003 TI - Identification and characterization of an auto-activating MEK kinase from bovine brain: phosphorylation of serine-298 in the proline-rich domain of the mammalian MEKs. AB - Mitogen-activated protein kinase kinases (MKKs or MEKs) are dual specificity tyrosine/threonine protein kinases that are activated by phosphorylation at two closely spaced serine residues (serines-218 and -222) by the c-mos and raf proto oncogenes. This double phosphorylation is both necessary and sufficient for MEKs to activate the MAP kinase enzymes in vitro. The specificity or regulation of in vivo signaling to the mammalian MEKs (MEK1 and MEK2) was recently reported also to involve the differential phosphorylation of a proline-rich peptide located between the MEK kinase-subdomains IX and X. Here we report the purification and characterization of an auto-activating protein kinase from bovine brain that phosphorylates serine-298 of the MEK1 and MEK2 proline-rich insert peptides. The auto-activation of the MEK-S298 peptide kinase is the result of an intermolecular phosphorylation event that can be prevented by the peptide substrates. The inactive kinase migrates on gel filtration as a 90 kDa protein, and after activation as a 43 kDa phosphoprotein. Incorporation of 32P[phosphate] into 40-42 kDa proteins on SDS-PAGE parallels the activation of the enzyme, and dephosphorylation by protein phosphatase 2Ac reverses the activation. SDS-PAGE renaturation assays show that the 40 kDa protein has the capacity to autophosphorylate, and exhibits kinase activity towards myelin basic protein after activation. Phosphorylation of purified bovine brain MEK or recombinant MEK1 by the auto-activated kinase does not activate the enzyme, and does not interfere with the in vitro raf-mediated MEK activation. We conclude that still unknown kinases may control the MAP kinase pathway by targeting MEK. PMID- 9416004 TI - Role of focal adhesion kinase in integrin signaling. AB - Integrins are the major cell surface receptors for extracellular matrix molecules, which play critical roles in a variety of biological processes. Focal adhesion kinase has recently been established as a key component of the signal transduction pathways triggered by integrins. Aggregation of FAK with integrins and cytoskeletal proteins in focal contacts has been proposed to be responsible for FAK activation and autophosphorylation by integrins in cell adhesion. This may be achieved by FAK interaction with talin or other cytoskeletal proteins that in turn associate with the cytoplasmic domain of integrin beta subunits. Autophosphorylation of FAK at Y397 leads to its association with Src, resulting in activation of both kinases. The activated FAK/Src complex acts on potential substrates tensin, paxillin and p130cas. Besides cytoskeletal regulation, FAK phosphorylation and/or binding to paxillin and p130cas may trigger downstream activation of MAP kinase by the adoptor protein Crk. Src association with FAK may also lead to its phosphorylation of other sites on FAK, including a binding site for Grb2. Cell adhesion-dependent association of FAK and Grb2 may provide a mechanism by which MAP kinase is activated in cell adhesion. PI 3-kinase has also been shown to bind FAK in a cell adhesion-dependent manner at the major autophosphorylation site Y397. This association could lead to activation of PI 3 kinase and its downstream effectors. Recent results from a number of different approaches have shown that integrin signaling through FAK leads to increased cell migration on fibronectin as well as potentially regulating cell proliferation and survival. PMID- 9416005 TI - Carbon and nitrogen sources regulate delta-aminolevulinic acid and gamma aminobutyric acid transport in Saccharomyces cerevisiae. AB - Evidence has been obtained showing that transport of delta-aminolevulinic acid (ALA), a precursor of porphyrin biosynthesis in Saccharomyces cerevisiae, is mediated by the gamma-aminobutyric acid (GABA)-specific permease, UGA4. In yeast GABA is also incorporated by the general amino acid permease (GAP1) and the specific proline permease (PUT4). The aim of the present work was to carry out a comparative study on the regulation of ALA and GABA transport to confirm our proposal that both compounds share the UGA4 permease. ALA and GABA uptake were measured in cells grown on minimal media with different carbon and/or nitrogen sources. To study the effect of the carbon source on UGA4 permease, ALA and GABA incorporation were measured in D27 strain, lacking GAP1 permease, and grown in proline as the sole nitrogen source, so the activity of PUT4 permease was negligible. The effect of the nitrogen source on UGA4 permease was studied measuring ALA and GABA uptake rates in cells from media with ammonium, proline and urea as nitrogen sources. It was found that the regulation by the carbon source was similar on ALA and GABA transport; they depend equally on the energetic conditions of the cells. Moreover, regulation by the nitrogen source on ALA and GABA uptake was also similar, and identical to that described already for UGA4 permease. These results are further evidence that both compounds, ALA and GABA, share the GABA-specific permease, UGA4. PMID- 9416006 TI - Ovine cardiac Na,K-ATPase: isolation by means of selective solubilization in Lubrol and the effect of 1 alpha,2 alpha-epoxyscillirosidin on this enzyme. AB - The inhibition of cardiac Na,K-ATPase by 1 alpha,2 alpha-epoxyscillirosidin is the principal cause of poisoning of cattle by the tulip, Homeria pallida. The ultimate goals of this study were to study the interaction between 1 alpha,2 alpha-epoxyscillirosidin and ovine Na,K-ATPase by means of inhibition and displacement binding studies. Ovine cardiac Na,K-ATPase was isolated in membrane bound form by means of deoxycholate treatment, high-speed ultracentrifugation, NaI treatment and selective solubilization in Lubrol. The inhibition of ovine cardiac and commercial porcine cerebral cortex Na,K-ATPase by 1 alpha,2 alpha epoxyscilirosidin and ouabain was studied using a discontinuous Na,K-ATPase assay. The binding of 1 alpha,2 alpha-epoxyscillirosidin, ouabain and digoxin to the above enzymes was compared using a displacement binding assay with [3H] oubain. The Lubrol-solubilized ovine cardiac Na,K-ATPase showed a specific activity of 0.3 U/mg with no ouabain insensitive activity. I50 values of 2.1 x 10(-8) and 2.7 x 10(-8) were obtained for the inhibition of this enzyme by 1 alpha,2 alpha-epoxyscillirosidin and ouabain, respectively. 1 alpha,2 alpha Epoxyscillirosidin has a much higher KD value (1.5 x 10(-7) M), however, than ouabain (9.5 x 10(-9) M) and digoxin (1.7 x 10(-8) M) in displacement binding studies with [3H]ouabain. 1 alpha,2 alpha-Epoxyscillirosidin is a potent inhibitor of ovine cardiac Na,K-ATPase and is a slightly stronger inhibitor of the enzyme than ouabain. The anomalous result for the displacement of 1 alpha,2 alpha-epoxyscillirosidin from its receptor is either a result of different affinities that K+ has for the enzyme ouabain and enzyme-1 alpha,2 alpha epoxyscillirosidin complexes or because of different complex stabilities of these complexes. PMID- 9416007 TI - Porphyrin-induced protein structural alterations of heme enzymes. AB - Some alterations in the protein structure of delta-aminolevulinic acid dehydratase (ALA-D) and porphobilinogen deaminase (PBG-D) induced by uroporphyrin (URO) and prototoporphyrin (PROTO) have been observed previously. To obtain further evidence of these phenomena, the absorption and fluorescence spectra of ALA-D and PBG-D and the total protein content of sulfhydryl and free amino groups were analyzed after exposure of the enzymes to URO I and PROTO IX, ALA-D and PBG D were partially purified from bovine liver and exposed to URO I or PROTO IX, both in the dark and under UV light. All experiments were performed in the enzyme solutions after removing the porphyrins. Absorbance spectra changes in the region of 220-300 nm were registered, indicating the interaction of the porphyrins with the molecular structure of the enzymes. The main changes in the fluorescence spectra were observed in the spectral region of 555 nm, and only slight modifications in the spectral region of 340-360 nm; moreover, alterations were stronger upon UV irradiation and in the presence of URO I when compared with darkness and PROTO IX. Variations in total SH groups would suggest the formation of disulfur bridges induced by URO I and the rupture of some S-S groups induced by PROTO IX. The effect of porphyrins on free amino groups would reflect a combination of cross-linking and fragmentation of proteins. Structural changes were observed when the enzymes were exposed to the porphyrin both in the dark or under UV light; however, they were stronger in the latter condition. These results suggest that porphyrins per se could act directly on the protein structure and that this action would be enhanced upon UV irradiation. PMID- 9416008 TI - Isolation and identification of porcine reproductive and respiratory syndrome virus in cell cultures. AB - Three strains of porcine reproductive and respiratory syndrome virus (PRRSV) were isolated in porcine lung macrophage (PLM) cultures from three swine herds. This has been the first successful isolation of PRRSV in the Czech Republic and the strains received the designations CAPM V-501, CAPM V-502 and CAPM V-503, respectively. All the three isolates in PLM were identified by immunofluorescence and immunoperoxidase tests and the strain CAPM V-502 also by electron microscopy using the ultrathin section technique. The strain CAPM V-502 has been adapted to the cell line MARC-145. Viral RNA in PLM cultures infected with any of the isolated PRRSV strains was demonstrated by RT-PCR targeted to the more conserved ORF 7 genomic region encoding the nucleocapsid protein. The assessment of PCR products in agarose gel revealed a uniform size of 394 bp in all the three isolates and the European prototype strain Lelystad used as positive control. PMID- 9416010 TI - Immunosuppression in dogs with pyoderma and/or demodicosis. AB - The occurrence of decreased activity of the immune system was studied in a group of 66 dogs with various combinations of pyoderma and demodicosis. Our complex examination of the dogs included the following: leukocyte count, differential count, phagocytosis, blastogenic lymphocyte transformation and quantitation of total serum immunoglobulins, lysozyme and haemolytic complement. Immunosuppression was found in 19 (28.8%) cases. Immunosuppression was rare in dogs with demodicosis and did not appear without a concurrent pyoderma. An increase in the neutrophil counts and total serum immunoglobulin levels significant was found in dogs with demodicosis combined with pyoderma. On the contrary, marked immunosuppression was detected in dogs with deep pyoderma. A considerable immunosuppression was present in 7 of 10 German shepherds dog pyoderma (GSP). Significant depressions were found in phagocyte activity and lymphocyte activity. Immunosuppression was observed in 4 of 9 dogs in other breeds with uncomplicated deep pyoderma. All groups of dogs with pyoderma showed a significant increase in total serum immunoglobulins. Conclusion from these findings is that deep pyoderma more than Demodicosis was concerned with immunosuppression. German shepherds with deep pyoderma had more expressed immunosuppression than other breeds. PMID- 9416009 TI - Effect of an adrenergic agonist and a cholinergic antagonist on the airway epithelium. AB - The ultrastructure of the rabbit tracheal epithelium was studied 30 minutes after intratracheal administration of two puffs of salbutamol and ipratropium bromide, respectively. The injury to the tracheal epithelium due to the treatment with both bronchospasmolytic drugs was considered moderate to severe. In both experimental groups, the degree of goblet cells' stimulation did not differ significantly, the ciliated cells were less damaged compared with the goblet ones and the morphological signs of the impaired self-cleaning ability were revealed. PMID- 9416011 TI - [Immunomodulatory effect of levamisole and administration of amitraz in dogs with uncomplicated generalized demodicosis]. AB - The immunomodulatory effect of levamisole (Decaris tbl.) in the course of acaricide therapy with amitraz (Taktic) on the functional activity of blood neutrophils (% of phagocytizing cells and ingestion capacity) and lymphocytes (blastogenic response to Con A) in dogs with uncomplicated generalized demodicosis (NGD) was studied. The level of examined parameters was evaluated before treatment, week 3 and 7 after the first application of these preparations; and compared with the values of NGD dogs treated only with amitraz and with those in clinically healthy dogs. In comparison with healthy dogs the initial level of examined activities of both cell populations was significantly depressed. A significantly earlier (4 weeks earlier) increase (when compared with values before treatment) of investigated activities of neutrophils and lymphocytes occurred in dogs treated with amitraz and levamisole in comparison with those in dogs treated only with amitraz. It was manifested especially significantly in phagocytosis, the ingestion capacity of neutrophils at this time of therapy has reached the level of those in healthy dogs. Functional activity of lymphocytes in both groups of NGD dogs has not reached a comparable value with that in healthy dogs either at the end of observation. The presented results indicate that significantly earlier improvement of functional activity of phagocytes and lymphocytes in demodectic dogs treated with amitraz and levamisole was connected with the immunorestorative effect of levamisole. PMID- 9416012 TI - Changes in stature following plyometric drop-jump and pendulum exercises. AB - The aim of this study was to compare the changes in stature following the performance of plyometric exercises using drop-jumps and a pendulum swing. Eight male participants aged 21.7 +/- 1.8 years with experience of plyometric training gave their informed consent to act as participants. Participants undertook two exercise regimens and a 15-min standing test in a random order. The exercises entailed the performance of 50 drop-jumps from a height of 0.28 m or 50 pendulum rebounds. Participants were instructed to perform maximal jumps or rebounds using a 'bounce' style. Measurements of stature were performed after a 20-min period of standing (pre-exercise), 2-min after exercise (post-exercise) and after a 20-min standing recovery (recovery). Back pain and muscle soreness were assessed using an analogue-visual scale, at each of the above times and also 24 and 36 h after the test. Peak torque during isokinetic knee extension at 1.04 rads-1 was measured immediately before and after the exercise bouts, to assess the degree of muscular fatigue. Ground/wall reaction force data were recorded using a Kistler force platform mounted in the floor for drop-jumps and vertically on the rebound wall for pendulum exercises. Drop-jumps resulted in the greatest (p < 0.05) change in stature (-2.71 +/- 0.8 mm), compared to pendulum exercises (-1.77 +/- 0.7 mm) and standing (-0.39 +/- 0.2 mm). Both exercise regimens resulted in a significant (p < 0.01) decrease in stature when compared to the standing condition. Drop-jumps resulted in significantly greater peak impact forces (p < 0.05) than pendulum exercises (drop-jumps = 3.2 +/- 0.5 x body weight, pendulum = 2.6 +/- 0.5 x body weight). The two exercise conditions both invoked a small degree of muscle soreness but there were no significant differences between conditions. Both exercise regimens resulted in a non-significant decrease in peak torque indicating a similar degree of muscular fatigue. Based on the lower shrinkage resulted and lower peak forces, it can be concluded that pendulum exercises pose a lower injury potential to the lower back than drop-jumps performed from a height of 28 cm. PMID- 9416013 TI - Dynamic cursor gain and tactual feedback in the capture of cursor movements. AB - Recent research involving a trackball with force feedback has demonstrated that tactile feedback can enhance the acquisition of targets in graphical user interfaces in terms of movement times and errors. The present study seeks to explore the degree to which tactual feedback over a target, in contrast to changes in the display/control gain over the target, influences target acquisition performance. Tactual feedback over a target is felt as a pulling force towards the centre of a target, with a counterforce applied when moving out of the centre. Changes in the cursor gain can be used to create a cursor-catching effect by requiring more movement effort of the control device to leave than to enter the target centre, without increasing the total amount of effort to enter and leave the target area. User movement in entering a target is thus braked by the change in cursor gain. Results of an experiment indicated that target acquisition performance was generally higher in the tactual feedback condition, followed by cursor gain feedback, in comparison with no-cursor gain feedback. User interface design issues as related to gain feedback in visual interfaces and tactual feedback over targets are considered. PMID- 9416014 TI - The effect of sustained spinal load on intra-abdominal pressure and EMG characteristics of trunk muscles. AB - Fourteen young male students (mean age 21 years, mean weight 69.4 kg, and mean height 175.4 cm) and 12 young female students (mean age 22.2 years, mean weight 60.6 kg, and mean height 169.3 cm) held 9.07 kg and 6.8 kg, respectively, at their three-quarters horizontal reach distance in upright and stooping postures for a period of 5 min. During these periods the external torque on the lumbosacral disc, intra-abdominal pressure, and electromyographic signals from erectores spinae at T12 and L3 levels, latissimus dorsi and external obliques were recorded at 1 kHz for 2.1 s every 15 s for a period of 5 min. The EMGs were processed in magnitude and time domains to determine muscle fatigue. Through the data obtained it was seen that the intra-abdominal pressure did not follow or reflect either the spinal load or the muscle activity. Based on the arguments presented, it was concluded that the intra-abdominal pressure does not appear to have a role of relieving the spine of some of its load. Instead, it is suggested that it is a dependent variable manifesting itself when mechanisms for spinal stability are evoked to overcome large voluntary and inertial loads. PMID- 9416015 TI - Force exertion in awkward working postures--strength capability while twisting or working overhead. AB - Isometric strength data have been collected for three of the awkward work situations often imposed by workplace constraints in industry, as found in jobs such as maintenance and repair. The effects of the task layout factors (location of the workpiece, reach distance and direction of force exertion) were investigated and the data has been tabulated to show the degree to which strength may be reduced in different situations. Strength measurements in the present study were found to be considerably higher than previous measurements, apparently due to differences in instructions on foot placement that were given to subjects, which indicates that even small constraints on posture within the workplace may have large effects on the ability to exert force. PMID- 9416016 TI - Strength capability while kneeling. AB - Work sometimes has to be carried out kneeling, particularly where jobs are performed in confined spaces as is common for miners, aircraft baggage handlers and maintenance workers. In order to assess the risks in performing forceful tasks under such conditions, data is needed on strength capabilities of kneeling subjects. A study was undertaken to measure isometric strength in single-handed exertions for male subjects and to investigate the effects on this of task layout factors (direction of force exertion, reach distance, height of the workpiece and orientation relative to the subject's sagittal plane). The data has been tabulated to show the degree to which strength may be reduced in different situations and analysis of the task factors showed their influence to be complex with direction of exertion and reach distance having the greatest effect. The results also suggest that exertions are weaker when subjects are kneeling on two knees than when kneeling on one knee, although this needs to be confirmed by direct experimental comparison. PMID- 9416017 TI - Congruence between parent satisfaction with nursing care of their children and nurses' perceptions of parent satisfaction. AB - The purpose of this descriptive study was to determine the degree of congruence between parents' satisfaction with nursing care on a pediatric neurosciences unit and nurses' perceptions of parent satisfaction. Convenience samples of 20 pairs (20 parents and 20 nurses) from the neurosciences unit participated in this study. Data were collected by means of a 25-item self-administered satisfaction with nursing care instrument and socio-demographic tools. Satisfaction with nursing care is a reliable predictor of overall hospital satisfaction (Abramowitz et al., 1987; Cleary et al., 1989). Satisfied health care consumers are known to show better rates of compliance with treatment regimens and be more willing to seek health care services (Greeneich et al., 1992; McMillan, 1987; Naylor, Munro, & Brooten, 1991). The findings of this research study support the need for nurses to explicitly ask consumers (patients and families) whether or not their expectations of nursing care are being met. For unless nurses ensure that the care they provide is consistent with what consumers want, nurses risk basing nursing interventions on assumptions and erroneous perceptions and consumers are unlikely to be satisfied. PMID- 9416018 TI - Living with dying: families coping with a child who has a neurodegenerative genetic disorder. AB - The impact on a family where a child has been diagnosed with a neurodegenerative genetic disorder is enormous. The anguish that accompanies parents from pre diagnosis to diagnosis to acute care to palliative care is tremendous. As well, the realization that a genetic disorder has affected one child, leads to the possibility that other family members may also be affected. Yet, given the amount of stress on most of these families, they are extremely resilient, and employ a variety of coping strategies to manage the situation. This paper focuses on the results of a questionnaire completed by 15 families. The questionnaire addressed the following research questions: 1) What are the unique features of the losses associated with a neurodegenerative genetic disorder?, 2) What are the coping strategies that families employ to manage the losses associated with a child who has a neurodegenerative genetic disorder?, 3) What support resources are required to better assist families to cope with a child who has a neurodegenerative genetic disorder?. Analysis of data revealed that families employ a variety of coping strategies to manage their day to day lives, and that these strategies changed with time and in relation to the child's condition. They also found that they required greater support during the first few months following diagnosis, however that once support services were in place, they felt confident in care for their child. The findings revealed that there are many unique aspects in caring for children with neurodegenerative genetic disorders that are different from other children with terminal illnesses. Implications for nursing and research are suggested. PMID- 9416019 TI - Certification concerns. PMID- 9416020 TI - An ongoing orientation for sponsored education activities. PMID- 9416021 TI - A report from the Nurses Certificate Program in Interactive Imagery. PMID- 9416022 TI - Discharge information needs and symptom distress after abdominal aortic surgery. AB - The purpose of this study was to describe the discharge information needs and symptom distress of people after abdominal aortic reconstructive surgery. Interviews (N = 51) were conducted prior to, and 4 weeks after, hospital discharge. People indicated that the most important information to help them manage their care after discharge related to the recognition, prevention and management of complications. Broken sleep and incisional pain were the most distressful of symptoms prior to hospital discharge, whereas fatigue and broken sleep were most distressful once home. These results may assist nurses to understand the discharge information needs and symptom distress of people recovering from aortic reconstructive surgery and the importance of discharge education to help people to manage their care once home. PMID- 9416023 TI - Women and heart attack: a study of women's experiences. AB - Cardiovascular disease in general, and myocardial infarction (MI) in particular, is the major health problem of females after 50 years of age. To date, heart disease research has focused primarily on males. The limited evidence suggests that the physical, psychological, and social ramifications of MI for women are significant, and different from those of men. Since the specific rehabilitation needs of women are not yet clear, this study was designed to explore the unique experiences and needs of women following a first MI. A phenomenological study using focus groups was used to explore the experiences, questions, concerns, and preferred interventions of women after a MI. Participants were female volunteers (n = 14) who had been hospitalized for a MI within the previous 6 months. Focus groups were audio taped and analysed by the investigators. Four major themes emerged: validation; perceived gender differences; role expectations/role tensions; and helps and hindrances to recovery. PMID- 9416024 TI - Patients evaluate an outpatient cardiac education program. PMID- 9416025 TI - Accessing patients, patient records, and patient databases: the "confidentiality access maze". Part II. PMID- 9416026 TI - Teamwork--the value that solidifies our unity of purpose. PMID- 9416027 TI - Protein: metabolism and effect on blood glucose levels. AB - Insulin is required for carbohydrate, fat, and protein to be metabolized. With respect to carbohydrate from a clinical standpoint, the major determinate of the glycemic response is the total amount of carbohydrate ingested rather than the source of the carbohydrate. This fact is the basic principle of carbohydrate counting for meal planning. Fat has little, if any, effect on blood glucose levels, although a high fat intake does appear to contribute to insulin resistance. Protein has a minimal effect on blood glucose levels with adequate insulin. However, with insulin deficiency, gluconeogenesis proceeds rapidly and contributes to an elevated blood glucose level. With adequate insulin, the blood glucose response in persons with diabetes would be expected to be similar to the blood glucose response in persons without diabetes. The reason why protein does not increase blood glucose levels is unclear. Several possibilities might explain the response: a slow conversion of protein to glucose, less protein being converted to glucose and released than previously thought, glucose from protein being incorporated into hepatic glycogen stores but not increasing the rate of hepatic glucose release, or because the process of gluconeogenesis from protein occurs over a period of hours and glucose can be disposed of if presented for utilization slowly and evenly over a long time period. PMID- 9416028 TI - Factors that influence the decision to receive treatment for proliferative diabetic retinopathy. AB - Educational messages aimed at health professionals have stressed the importance of regular eye examinations for people with diabetes and the value of early treatment. To investigate whether the messages need to be expanded or tailored in a specific way, we asked people with diabetes (N = 37) to describe their reactions to the diagnosis of proliferative diabetic retinopathy and the factors that influenced their decision to seek treatment. The findings reaffirm the importance of the physician's recommendation in pursuing treatment. Furthermore, a constructive response (eg, "knew it had to be taken care of") to the diagnosis was associated with prior knowledge of the consequences of proliferative diabetic retinopathy. Minimal racial and gender differences were observed. White women reported being influenced by the experiences of others, and African Americans reported being influenced by the diabetes educator. These findings emphasize the importance of providing patient education not only following a diagnosis but also in anticipation of probable complications. PMID- 9416029 TI - Detecting vascular problems in patients with diabetes treated with an intra aortic balloon pump. AB - Individuals with diabetes are at increased risk for both peripheral vascular disease and coronary artery disease. In patients with severe coronary artery disease, a cardiac assist device called an intra-aortic balloon pump (IABP) often is used to aid the failing heart and prevent further cardiac ischemia. Because this device is inserted via the femoral artery, patients are at risk of limb ischemia distal to the insertion site. Patients with diabetes are particularly prone to this complication. Detecting the early signs and symptoms of ischemia is crucial to preventing serious sequelae. Standard vascular examination techniques, in addition to being subjective and not easily reproducible, may be misleading in patients with diabetes. This article provides a review of the signs and symptoms of lower limb ischemia and noninvasive vascular tests that clinicians can use to evaluate lower extremity circulation. Also included are protocols for patient care during and after hospitalization, and two case studies of cardiac patients with diabetes who were treated with an IABP. PMID- 9416030 TI - A clinical path for adult diabetes. AB - The use of clinical paths for patient care management was explored by this development team as a mechanism to provide consistent, high-quality care to hospitalized patients in high-volume, high-risk diagnostic categories. Reviewing the historical aspects and importance of clinical paths helped expand the team's perspective to incorporate pre- and posthospitalization phases of patient care into the clinical path being developed. A multidisciplinary team of physicians, nurses, health educators, and dietitians from both inpatient and outpatient departments of Kaiser-Santa Teresa Medical Center in San Jose, California, devised and implemented an Adult Diabetes Mellitus care path. Staff education preceded the implementation of the care paths. Measurements of quality indicators showed improvements in patient satisfaction, patient education, patient knowledge, and nutrition assessments. PMID- 9416031 TI - Moving diabetes management from clinic to community: development of a prototype based on automated voice messaging. AB - The purpose of this study is twofold. First, it provides a review of the literature supporting the development of a new service to help patients with diabetes and their providers manage their care. This service, automated voice messaging (AVM) with nurse follow-up, allows for systematic and intensive patient monitoring and diabetes education as well as a means of focusing clinical resources where they are most needed. Second, it provides a description of a prototype AVM-based diabetes management service that has been developed as part of two ongoing, randomized, controlled trials to test the efficacy of AVM care for patients with Type 2 diabetes. Preliminary findings from implementing this service in two large public healthcare systems suggest that AVM-supported care is feasible, desirable by clinicians and patients with diabetes, and may identify serious health problems that otherwise would go unnoted through standard means of clinic-based patient care. PMID- 9416032 TI - A mentoring program for adolescents with diabetes. AB - Adequate psychosocial support is a major factor in well-managed diabetes, especially with newly-diagnosed adolescents who face many life changes. A review of the literature shows that few psychosocial support systems exist for adolescents with diabetes. Few psychosocial interventions have been tested and shown to be effective in improving the diabetes-related behavior of adolescents. The purpose of this paper is to provide an overview of the emotional and developmental needs of adolescents who are newly diagnosed with insulin-dependent diabetes mellitus and to propose a mentoring program that pairs a qualified, supportive young adult who is knowledgeable about diabetes with a newly diagnosed teenager with a similar socioeconomic background. The trained mentor will provide support to the adolescent regarding diabetes-related issues as well as other issues related to adolescence. PMID- 9416033 TI - Diabetes self-management education programs in the Veterans Health Administration. AB - The role of diabetes educators is likely to evolve and expand due to the development of chronic disease management programs that are based on a multidisciplinary approach of healthcare providers. In addition to providing self management education and training to more patients with diabetes, educators will most likely need to expand their services to include case management and quality management of process and health outcomes. PMID- 9416035 TI - Menopausal transition of Korean immigrant women: a literature review. AB - The menopausal transition needs to be understood in terms of the multiple mediating factors within the context in which women experience it. For immigrant women especially, the menopausal experience is complicated by multiple transitions and social marginality, so it cannot be adequately explained without considering this complexity. In this paper we review the literature on the menopausal transition of a group of vulnerable immigrant women in two ways: describing the transitions themselves (menopause, immigration, and housewife to employee), and describing factors that mediate the menopausal transition experience (family norms, meaning of menopause and women's work, and health practices). We emphasize the context and suggest areas of needed research. PMID- 9416036 TI - A biographical disruption: the case of an abnormal pap smear. AB - In this study we explore the biographical disruption resulting from a diagnosis of an abnormal Pap smear and the consequent process of biographical reconstruction. This is a qualitative study of thirteen women between the ages of 19 and 54 years who were diagnosed with an abnormal Pap smear and underwent colposcopy treatment. Data collection was through individual in-depth interviews, which were transcribed and analyzed by a team of researchers for important themes. An opportunistic sampling strategy was used. The inherent ambiguity in the diagnosis, its treatment strategies, the prognosis of their condition, and patients' fear of cancer all made the process of biographical reconstruction more problematic. By putting their faith in medicine and mobilizing their personal resources, women attempted to reestablish a positive personal and social self. Further exploration of the long-term psychosocial impact of this diagnosis is warranted so that women's emotional as well as medical needs are adequately addressed. PMID- 9416037 TI - Menstrual and premenstrual experiences of women in a developing country. AB - The menstrual and premenstrual experiences of black Zimbabwean women were examined. Twenty-five professional women and twenty-five domestic workers were interviewed using semistructured, open-ended interviews designed to explore their experiences. An analysis of consensual data indicated a number of predominant themes, including secrecy associated with menstruation, the negativity surrounding menarche, the breakdown of the traditional family network that passes on information about menstruation, and the acceptance of menstrual cycle experiences by the women. The main differences between the two groups concerned explanations of the functions and purpose of menstruation and the reporting of physical and affective symptoms. These differences are suggestive of the impact of educational level on experiences of menstruation. PMID- 9416038 TI - Violence against ex-wives: anger and advocacy. AB - Researchers on violence against women have failed to reveal any studies that provide definitive explanations for the violent behaviors of certain males. However, violence against women has been known to occur as a result of unusually potent situational stressors, regardless of the male's propensity toward violence. Some of these situational stressors occur during the process of marital separation and divorce, particularly in relation to disputes over child custody, support, and access. In this paper I report on a second-level analysis of a set of studies in Australia aimed at examining the experience of separating parents who did not gain custody of their children. One of the dominant themes that emerged in the initial analysis from the male cohort was the real or vicarious violence that pervaded the men's interactions with their ex-spouses. These men, from a range of socioeconomic backgrounds and age groups, freely discussed episodes in which they had either planned, executed, or fantasized about violence against their spouses in retaliation for real or perceived injustices related to child custody, support, and/or access. In many cases, these thoughts and actions were reinforced by the encouragement of other males. To a lesser extent, women also reported violent inclinations induced by the situation. The implications for those advocating for women and families are clear. There is a need to understand the experience of marital separation from the perspective of both spouses as a basis for family counseling. We must also heighten awareness of the need to educate young people away from an "ownership" model of marriage and relationships, which is counterproductive to the personal development of both partners. Equally important is the need for all health professionals to advocate for changes that would help to correct injustices in the family court system, many of which are related to gender issues. PMID- 9416039 TI - An analysis of menopause in the popular press. AB - With the increased interest in menopause, it is imperative that midlife women be given reliable information. An analysis of the lay literature on menopause over the past ten years in the United States was conducted in order to determine the content of the articles, their source, and their credibility. Most articles were found to blend opinion with fact, and many of the authors of these articles did not have any stated qualifications in midlife women's health. Physicians, particularly those espousing the medicalization of menopause, were the most often quoted experts, and few nurses or other health care workers were cited as experts. It is suggested that nurses become more involved in health education for midlife women. PMID- 9416040 TI - Old skills and new knowledge: midwifery in contemporary Zimbabwe. AB - Sixty-one traditional birth attendants residing in the southern sector of Zimbabwe were interviewed concerning their midwifery practice. Traditional midwives were interviewed individually to gather information concerning: (a) the development of traditional midwifery skills, (b) the nature of traditional birthing patterns, (c) the features of the one-week midwifery training program provided to upgrade traditional midwives, and (d) traditional midwifery as practiced today, post the one-week training program. In describing past and present traditional midwifery, they reported a change in the use of sanitation practices, a heightened understanding of at-risk pregnancies and the need for formal medical intervention, and the adoption of mechanisms to record new births. PMID- 9416041 TI - Recontextualizing sexuality in chronic illness: women and interstitial cystitis. AB - Interstitial cystitis (IC) is a chronic bladder condition that affects ten times more women than men. The major symptoms of IC include urinary frequency, urgency, and suprapubic pain. Dyspareunia is another common complaint. The construct of sexuality can best be understood in context, from the perspective of the study participants. To better understand the changes in sexuality experienced by women with IC, the findings from a survey of 138 women with IC are compared with responses from an earlier pilot interview with ten women with IC. Managing any chronic illness requires changes in many aspects of one's life, including activities, self-perceptions, relationships, and roles. Women with IC described what was important to them "as women" in the context of their current primary life values of health, love, and family. Changes in the sexual relationship were described in the context of the effects of having IC and the centrality of maintaining relationships. The healing potential of participation in support groups is discussed in relation to the women's desire to maintain independence and to help others with the disease. PMID- 9416042 TI - Women's experiences of barriers to support while caregiving. AB - This qualitative study explored women's perceptions of barriers to support during family caregiving in a Canadian setting. Twenty mothers of premature infants and twenty women caring for an older person who is cognitively impaired were interviewed in-depth over 18 months. Both groups of women preferred that support be offered to them and identified numerous barriers to requesting support. Perceived barriers included an obligation to provide care, loss of independence and self-esteem, concern for burden on others, the desire to excuse others from providing support, the inability to reciprocate support, fear of refusal or exposure, nonsupportive actions, the time and effort needed to coach others to provide effective support, and the lack of available, competent help. Some of these barriers reflect personal costs to the woman caregiver. Other barriers reflect societal norms that family caregiving is the responsibility of women. PMID- 9416043 TI - Value conflict: the lived experiences of women in treatment for weight loss. AB - My purpose in conducting this phenomenologic study was to describe how women in weight treatment integrated the requirements of their programs into their daily life activities and the meaning they attached to this experience. Feminist theory was used to guide the inquiry and to interpret the findings. The central theme of value conflict was evident in five of the six women interviewed. The value conflict involved the tension between spontaneity and control in the lifestyle patterns of the participants. These women's experiences were characterized by a sense of struggle as they sought to balance the dietary restrictions and exercise requirements of their programs with emergent priorities. A conclusion drawn from the study was that current weight treatment programs have a less than optimal fit with women's values and needs. The importance of developing weight treatment models that promote women's autonomy in evaluating and managing their health is emphasized. PMID- 9416044 TI - Factors affecting graduating nurses' nutritional knowledge: implications for continuing education. AB - Nurses over 25 years of age and with more than 10 years of health-related work experience scored highest when tested for nutritional knowledge in a study of 129 midwestern graduating nurses. Significant differences (F [2, 128] = 5.8, p = .02) were revealed between nurses with 16 or more hours of clinical practice per week in comparison to those nurses with less than 16 hours of clinical practice per week. Cultural nutrition content was the area in which graduating nurses scored the lowest while food and nutrition policies/regulations was the area with which nurses were most familiar. These results have implications for hiring graduating nurses who will need the necessary nutritional care skills to provide quality nursing care today. PMID- 9416045 TI - Refresher courses for the 21st century. AB - One hundred eighty-one (n = 181) nurses who completed the Rutgers Nurse Refresher course between the years of 1991 and 1994 were surveyed to determine the outcome of the course in relation to successor return to nursing practice. Of the number of nurses who responded (n = 111), 78 nurses (70.2%) returned to practice as RNs. Nurses who completed the course in 1991 and 1992 had higher employment rates as RNs than those nurses who completed the course in 1993 and 1994. Of the 78 nurses who found employment as RNs, 55 nurses (71%) found employment in other-than hospital settings. An increase in employment in long-term care and other settings, such an physician offices and school nursing was found when the data were analyzed by setting and year of course completion. Findings of the survey are discussed in relationship to changes in health care delivery as well as recommendations for curriculum revision. PMID- 9416046 TI - Nursing education and nursing research utilization: is there a connection in clinical settings? AB - According to a survey of 753 nurses in Northern California, nurses with higher levels of education are more involved in research utilization and research activities. The data support the American Nurses' Association model of progressive involvement in research utilization and research conduct. Recommendations to improve research utilization in clinical and academic settings are provided. PMID- 9416047 TI - Comparing the effectiveness of two methods of delivering continuing education to school nurses. AB - On-site continuing education opportunities for school nurses are limited due to school nurses' geographic dispersion, yet the need is great. This study was designed to create, deliver, and evaluate two methods of providing a continuing education course on comprehensive school health programming for middle school and high school nurses: a 4-day on-site workshop and a six-module home-study course. The home-study course was less expensive to deliver than the workshop, but participants were less likely to complete it. The 4-day workshop had a higher rate of completion and led to greater networking and a stronger sense of self efficacy among participants. PMID- 9416048 TI - Nurse manager orientation: guidelines to meet the challenge of a rapidly changing role. AB - A management development program at The Johns Hopkins Hospital in Baltimore addresses the competency needs of 68 nurse managers. Orientation of new nurse managers is a significant component of the program. The orientation is based on performance standards and identification of organizational resources and opportunities to assist the managers to meet the standards. A convenience sample of 10 nurse managers, assistant directors, and directors of nursing reviewed the program 1 year after implementation and found it had improved the quality of orientation by providing structure and organization. The model can be applied to other institutions and practice roles. PMID- 9416049 TI - Student perceptions of academic advising in an RN-to-BSN program. AB - BACKGROUND: This study evaluated academic advisement from the perspectives of RN to-BSN students in relation to the advisement elements of function, style, and outcome. METHOD: Using multistage sampling, data were collected from 323 students enrolled in either a beginning, mid-way, or final course in the RN-to-BSN curriculum. RESULTS: All aspects of advisor style were perceived to have been employed and important. None of the outcomes were perceived to have been achieved nor important. There was a significant difference (p < 0.05) between students from ADN and Diploma educational backgrounds in relation to their perceptions of the importance of advisement functions and outcomes. Students rated their perceptions in accordance with andragogical learning principles that stressed the importance of individuality. The researcher developed Academic Advisement Questionnaires have content validity and reliability for internal consistency at 0.94 by Cronbach's coefficient alpha. CONCLUSION: Advisement in RN-to-BSN educational programs can positively influence student retention rates by providing advisement that is conducive to the needs and values of adult learner students. PMID- 9416050 TI - Assessing the elderly for infections. PMID- 9416051 TI - Occupational health and the the provision of nursing care for older adults. AB - Providing nursing care to the senior citizen is a rewarding and exciting area of nursing. Care is provided in a vast array of settings from home health and day care to extended residential care sites. These challenges require health care workers to be adaptive and innovative, attentive to detail, compassionate teachers, and promoters of safety for their patients and for themselves. An understanding of the transmission of microorganisms, knowledge of protective mechanisms, and a focus on safety will help ensure good health for the nurse and the elder client. Maintenance of occupational safety and health are combined efforts for the employee and the employer working together to ensure a safe environment for those who need services and those who provide them. PMID- 9416052 TI - Social barriers to recognizing HIV/AIDS in older adults. PMID- 9416053 TI - Understanding Creutzfeldt-Jakob disease. AB - The "mad cow disease" that decimated cattle in England has brought wide-spread attention to a similar disease in humans, Creutzfeldt-Jakob disease (CJD). This has led to concerns about the transmissibility and contagiousness of the infectious agent from possible CJD patients to health care workers and family members. Despite these worries, the occurrence of CJD in the human population has remained stable over the years at an incidence of about one person per million in the United States population, and increasing to six individuals per million for the older-than-60 age group. This article will review the etiology, clinical manifestations, and potential for transmission of this unusual infectious agent. PMID- 9416054 TI - Environmental rounds. What matters for infection prevention and control. PMID- 9416056 TI - The use of support groups in an adult day health center. PMID- 9416055 TI - California skilled nursing facilities. AB - It is estimated that of the 2.2 million people who turned 65 years of age in 1990, approximately 1 million will be cared for in a SNF at least once before they die (Satterfield, 1993). Under pressure from federal and state governments to reduce costs associated with acute care hospitalization, SNFs are admitting patients who require more complex medical and surgical care than in the past. Until recently, FS-SNF ICPs did not have the motivation to develop and implement complex infection prevention and control programs because their patients required a level of care that assured that their activities of daily living were met. In today's continuum of health care structure, SNFs are expected to maintain patients on ventilators and hyperalimentation and to care for patients recovering from complicated postsurgical procedures, such as total hip replacements and coronary artery bypass grafts. These patients are not expected to remain in SNFs until they die. They are expected to recover to their prehospitalization health status and, at a minimum, be discharged to the next lower level of care (i.e., board and care homes) or to assisted care in the family home. With the increased level of complex medical and surgical care that SNFs are now required to provide, infection prevention and control programs take on new emphasis. This is evidenced by the educational needs assessment reported by Leinbach and English (1995) and confirmed by this study. The basic principles of infection prevention and control are the same whether the patient is in a hospital, a DP-SNF, or a FS-SNF. However, the person responsible for infection prevention and control for a DP-SNF may be the same individual who is the ICP for the acute care hospital of which it is a part. Persons designated as ICPs for FS-SNFs often function in a much more isolated setting with limited access to resources. It is now time for experienced hospital-based ICPs to reach out to personnel in FS-SNFs to work together to develop and implement effective infection and prevention and control programs that meet the needs of patients cared for across the continuum. PMID- 9416057 TI - Holistic health interventions. The same difference. PMID- 9416058 TI - Changing nurses in a changing environment: saving your profession. PMID- 9416059 TI - Changing roles for psychiatric clinical nurse specialists: prescriptive privileges. AB - Interest was expressed at the mental health center in expanding the role of psychiatric clinical nurse specialists to include prescribing medications. There is a definite need for this expanded prescriptive role. The goals of improving patient access to care, offering quality service and cost effectiveness can be met by the psychiatric clinical nurse specialist certified by ANA. The functions of a CNS in a collaborative role with psychiatrists would involve promotion and health maintenance, evaluation, intake screening, health teaching, community action, and advanced psychobiological interventions including the prescribing of pharmacological agents. The ARNP, CS could alleviate the case load for pharmacological evaluation for the psychiatrists and enhance access to pharmacological management. The change in the Kentucky law giving prescriptive privileges to ARNPs makes this role feasible. PMID- 9416060 TI - KNA nursing history. PMID- 9416061 TI - Interviewing as a teaching strategy for promoting cultural sensitivity. AB - Interviewing was found to be an excellent teaching strategy in the promotion of cultural sensitivity in the course described in this article. Students were active learners in the acquisition of knowledge and critical thinkers in the analysis of the knowledge. Leininger (1991) emphasized the need for cultural sensitivity and cultural specific care in assisting supporting, facilitating, and/or enabling "individuals or groups to maintain or regain their well being (or health) in culturally meaningful and beneficial ways, or to help people face handicaps or death" (p.47). The need for further research on teaching strategies for promoting cultural sensitivity and on interviewing as a teaching strategy was identified. PMID- 9416062 TI - Collective bargaining for nurses anomaly or trend? PMID- 9416063 TI - Change in the nursing workforce: our views and suggestions for dealing with it. PMID- 9416064 TI - Nurses need collective action to 'build' profession. PMID- 9416065 TI - A person who is sick deserves the chance to get well. PMID- 9416066 TI - Nichols urges nurses to review, rethink future. PMID- 9416067 TI - Curtin questions 'new order'. PMID- 9416068 TI - A tribute to black male nurses. PMID- 9416069 TI - The 1997 legislature in review. PMID- 9416070 TI - Planning for adult learning. PMID- 9416072 TI - Peer assistance announces formation of four new nurse support groups. PMID- 9416071 TI - Medicare and advanced practice nurses. We did it! PMID- 9416073 TI - New Jersey Prescription Blank information update. PMID- 9416074 TI - The future of clinical nurse specialists in psychiatric-mental health nursing. PMID- 9416075 TI - Defining professional needs and goals: what do I want out of nursing? AB - The role of the clinical nurse specialist in psychiatric-mental health nursing is undergoing transformation secondary to massive changes occurring within the health care delivery system, as well as the larger society. In order to address these changes and their influence upon the evolution of the advanced practice role of clinical nurse specialists in psychiatric-mental health nursing, an Ad Hoc Committee consisting of twelve members of The Society of Certified Clinical Specialists in Psychiatric Nursing of the New Jersey State Nurses' Association met in 1996 and 1997 in order to explore and define the future of clinical nurse specialists in psychiatric-mental health nursing. PMID- 9416076 TI - Institute for Nursing/New Jersey State Nurses Association. Guidelines for research grant proposals. PMID- 9416077 TI - NISNA working to change board of nursing process for impaired nurses. PMID- 9416078 TI - Milking the wound. PMID- 9416079 TI - Computer use in ADN programs. PMID- 9416080 TI - Toys "R" clinical. PMID- 9416081 TI - Let's play nurse. PMID- 9416082 TI - The NCLEX-RN examination is changing: are you ready? AB - The NCLEX-RN Test Plan has been revised. The test plan retains "Client Needs" as the major content dimension and integrates "Phases of the Nursing Process" into all areas. Client Needs are divided into four categories. The four categories are divided further into a total of 10 subcategories that will be used to determine the percentage of questions on each candidate's examination. The authors describe the changes to the NCLEX-RN Test Plan and implications for nursing educators. PMID- 9416083 TI - Teaching tools. A simulated clinical problem-solving experience. PMID- 9416084 TI - Mentorship and professional role development in undergraduate nursing education. AB - Mentoring is a positive method for promoting professional development; however, there is little information in the literature addressing the use of mentoring with baccalaureate nursing students. Because professional socialization is a critical component of baccalaureate nursing education, mentoring would be an optimal methodology. The authors describe how a mentorship program combined with a professional role development course was integrated into a BSN nursing program. The evolution of the faculty-student mentorship program and the professional role development course is presented. PMID- 9416085 TI - Student-selected clinical assignments. PMID- 9416086 TI - Teaching nursing students evidence-based nursing. AB - Nurse educators face many challenges in the current healthcare environment. Educational methods, philosophies, and the content of curricula need to be reexamined to meet the needs of professional nurses who will practice in the next millennium. Evidence-based nursing is one approach that may enable future healthcare providers to manage the explosion of new literature and technology and ultimately may result in improved patient outcomes. The authors provide an introduction to evidence-based nursing as well as a description of the process in two separate undergraduate nursing programs. PMID- 9416087 TI - Ensuring success. The Faculty Development Plan. AB - In higher education, faculty advancement is based on demonstrated productivity in scholarship and service as well as teaching. In nursing, these expectations, added to clinical obligations, can make new faculty feel overwhelmed. The author recommends that new faculty take responsibility for their professional development by working with a dean or department chair to design a 5-year Faculty Development Plan that includes mentoring and participating in other campus support networks. PMID- 9416088 TI - Developing outcome assessment methods. AB - Describing the development of an outcome assessment method for accreditation, the authors discuss development of criterion definitions and the evaluation of results from a pilot study using student portfolios. The subsequent design of an outcome assessment tool to provide concrete, objective data, the evolution of the evaluation process using this tool, and how the outcome data are currently used for curriculum evaluation are discussed. PMID- 9416089 TI - Clinical learning experiences of nontraditional age nursing students. PMID- 9416090 TI - A method for developing faculty leaders. AB - The authors explore peer review as a mechanism for developing faculty leaders who can meet the educational challenges posed by public demands for accountability in higher education and an increasingly diverse student body. How educational paradigms influence the design of peer review programs is described. Incentives and disincentives as well as options for counteracting disincentives also are considered. Recommendations include the need to study effects of peer review programs on productivity, professional relationships, and educational outcomes within situational contexts, resources, and constraints. PMID- 9416091 TI - The 30-second women's health-promotion message. An innovative classroom strategy. PMID- 9416092 TI - ROPES: an experiential learning activity for leadership skills. AB - The cluster of activities called ROPES is borrowed and modified for teaching from OUTWARD Bound, the renowned outdoor management training program. ROPES uses experiential learning situations in which characteristic problem-solving tasks encourage the participants to stretch their personal limits and to learn teamwork. In doing so, trust, self-confidence, and communication and leadership skills develop through accomplishment of specific challenging activities. ROPES activities were found to be an effective method of instruction for teaching leadership in a senior nursing leadership course. PMID- 9416093 TI - All the efficiency of the U.S. Postal Service along with the compassion of the Internal Revenue Service. PMID- 9416095 TI - Cost-effectiveness of clustered unit vs. unclustered nurse floating. AB - This study evaluated costs and staffing balance outcomes comparing unrestricted unit floating (UUF) with cluster [by related patient population or technical requirements] unit floating (CUF) practices. Researchers used a computer simulation model with data from a 400 bed VA hospital. Literature suggested a high nurse turnover rate associated with dissatisfaction engendered by forced floating to unfamiliar units. Direct wage cost differences were negligible when UUF and CUF floating patterns were compared, so absolute costs were not the defining issue. UUF staffing patterns produced significantly fewer understaffed shifts (by nursing hours) than CUF floating permitted. The essential quality of care trade-off is between the UUF pattern that provided sufficient nursing hours of care vs. the CUF pattern that provided less absolute availability in hours of nursing care, but a better oriented staff. The author suggests seeking staff input when deciding which of these two floating patterns would be most acceptable in a particular institution. PMID- 9416094 TI - Process leadership and the death of management. AB - New information technology infrastructure allows point-of-service knowledge workers to make timely, informed decisions and take action without the traditional manager's close oversight or the top-heavy hierarchical structure associated with vertically managed organizations. Process leaders serve as consultants, coaches, linkers, and integrators to support front-line decision makers in a horizontally oriented model. The leaders must then be able to invest control, authority, and accountability for outcomes in the hands of the team. The process leaders, however, must be able to integrate resource information and generate knowledge and support for the providers of care that will enable them to make effective decisions about the requisite work. Process-oriented design in health care organizations emphasizes the interdependence of nurses, physicians, therapists, and technologists who must share the decisions and the risks. Thus, effective and efficient operations depend on recognizing the need for improved communications among team members and respect for the contributions of each. PMID- 9416096 TI - Supporting excellence in turbulent times. AB - Re-engineering offers an organization the opportunity to identify and strengthen critical system values and enhance employee commitment despite needed downsizing and major role changes. Anticipated distress during turbulent transitions can be decreased by managers paying close attention to employees' realistic concerns and offering opportunities for discussion about unaltered values as well as concretizing operational guideposts during the "in between" phase. Leaders must develop significant new skills to help staff develop competencies related to change management. Chief among the needed capabilities are direct communication techniques, enhanced self knowledge and desired team role behaviors. The expanded span of accountability for managers offers opportunities for staff empowerment through their needed participation in peer reviews, as well as the concomitant changes in position descriptions, role expectations, and financial rewards. PMID- 9416097 TI - Managed care organizations' arrangements with nurse practitioners. AB - Thirty-four of 67 MCOs in New York and Connecticut responded to requests for information on the roles, participation, and listing of nurse practitioners as primary care providers or in other capacities. MCO executives report a high degree of satisfaction with NPs who serve as their primary care providers, especially in women's health and geriatrics, as they spend more time teaching and explaining procedures than physicians. Ongoing lack of up-to-date information and/or confusion about the scope of NP practice exists among both health care professionals and the public. Perceived differences in the scope of care provided by NPs was related to state regulations, physician practice patterns, and availability of primary care physicians. Eighty-five percent of MCO executives thought their organizations should encourage the use of NPs. PMID- 9416098 TI - Protecting health care consumers: a Bill of Rights and responsibilities. PMID- 9416099 TI - Workforce reductions: low morale, reduced quality care. AB - As the number of positions decreases, the workload becomes more stressful for nurses left to pick up the slack. Mistakes are made, patient complaints increase as tensions rise, and the quality of nursing care decreases. The use of contingency staffing and overtime may increase as the workforce is reduced. Lack of job security forces acceptance of overtime, leaving less time for family life which may lead to resentment. The success of an organization is linked to employees' willingness to perform and use their skills. With deteriorating attitudes, employees will not perform at maximum effectiveness. Services do not meet established standards or customer expectations and are reflected in negative customer feedback and decreasing revenues. "There are no quick fixes. Tossing out last month's 'cure' to usher in this month's idea is a big waste of time" (Austin, 1994, p. 19). The impact from layoffs has long-lasting effects on employees, their families, and the community. Support for those displaced, and for those retained, provides a release for pent-up emotions and allows employees to get on with the work at hand. Workforce reductions will continue with the decrease in funding and the decline in patient census, but it is imperative that the quality of care be maintained. Registered nurses cannot be replaced at the bedside by UAP who do not have the specialized knowledge and skills required to provide safe and effective care (Thomas, 1995). Efforts to cut costs should be directed toward decreasing waste and eliminating redundant work, not at decreasing the number of RNs. The RN must remain the primary caregiver at the bedside to maintain quality care. Changes that remove the RN from the bedside will influence the quality of care that patients receive in the future. Increased demands and fewer, less-experienced staff result in less time for patient care. One negative patient outcome can be much more costly, directly and indirectly, than the salaries of several staff nurses. PMID- 9416100 TI - Clinic report cards: performance improvement reports in ambulatory care. PMID- 9416101 TI - Balancing personal and professional responsibilities. PMID- 9416102 TI - The people side of transformations. AB - All leaders in health care today are charged with the responsibility of transforming present practices into new and different ones that are needed for the future. The structural side of transforming is ultimately easier than the human side. However, the most frequent failures come from not concentrating sufficiently on the behavioral side of the change. Structural and psychological change must occur simultaneously and embrace each other for best results. PMID- 9416103 TI - Ethical problems in health care. PMID- 9416104 TI - Patients' rights in laboratory examinations: do they realize? AB - This article discusses the rights of patients who are attending hospital for the most common laboratory examinations and who may also be taking part in research studies. A distinction is made between five kinds of rights to: protection of privacy, physical integrity, mental integrity, information and self determination. The data were collected (n = 204) by means of a structured questionnaire specifically developed for this study in the clinical chemistry, haematological, physiological and neurophysiological laboratories of one randomly selected university hospital in Finland. The analysis of the data was statistical. On the whole, patients' rights were realized reasonably well. This was most particularly the case with protection of privacy, as well as with the rights of physical and mental integrity. The rights to information and self determination were less well realized. There are various steps that health care professionals and organizations can take to make sure that patients can enjoy their full rights, by counselling the patient, by giving opportunities to plan the examinations in advance, and by arranging a sufficient number of small examination rooms. PMID- 9416105 TI - Using clients. AB - An important part of the student nurse's training involves reflection on practice, as expressed in written assignments and seminar discussions. In this, students make use of material drawn from their work with clients. A key ethical question is, therefore: should clients' permission be sought by students for this use of case material in coursework assignments. This article examines in some detail the arguments both for and against seeking clients' permission and concludes that, in view of the principle of respect for autonomy, there is a moral obligation to gain consent. It is argued, however, that there may be legitimate exceptions to this. PMID- 9416106 TI - Moral sensitivity in psychiatric practice. AB - This study reports the results of a study of Swedish psychiatrists' responses to moral statements related to decision making in the psychiatric context. Use was made of the Moral Sensitivity Questionnaire, a modified instrument previously constructed from a theory of moral sensitivity. This Likert-type scale contains 30 items constructed from the following categories: interpersonal orientation, structuring moral meaning, benevolence, modifying autonomy, experiencing moral conflict, and trust in medical knowledge and principles of care. The purpose was to identify possible differences in responses rather than to evaluate right or wrong responses. The analysis is based on 754 completed questionnaires. The results of the study showed some significant differences in the item and category levels; for example, male psychiatrists experienced more conflicts than female psychiatrists and agreed to a greater extent that medical knowledge was most important in deciding what was best for the patient. The results also showed that more female than male psychiatrists thought that the relationship with the patient was most important in psychiatric practice. PMID- 9416107 TI - A model for conceptualizing the moral dynamic in health care. AB - Ethics involves an organized, reasoned approach to gathering and processing data in order to arrive at decisions about what to do, what to value, and/or what virtues to cultivate. A model is proposed for conceptualizing this complex dynamic, which incorporates elements of both rule-and-principle ethics and the ethic of care. The model suggested here has two levels. The first level identifies the components that comprise philosophical reasoning; the second contextualizes and operationalizes the model in relation to the processor's philosophical stance on the nature of knowing. Three philosophical stances are identified and described: science-dominant, person-dominant, and science-person equilibrium. Physicians tend to process patients from first perspectives, nurses from second. Hence, health team collaboration in moral problem solving is critically important. PMID- 9416108 TI - Nursing: a spiritual perspective. AB - This article explores and examines the fundamental need for nurses to include the promotion of the spiritual dimension of the health of human beings as well as the physical, mental and social facets if they truly wish to engage in holistic care. The author attempts to define the phenomenon of spirituality, aware of the dilemma that many individuals face when thinking and reflecting on this very personal and intangible issue. To be spiritual is to become fully human, the article argues, and the reverse is also true. Spirituality in health is inextricable in each person's search for the discovery of the truth about self and the meaning and purpose of life. Healthy communities are the product of healthy individuals who sow spiritual seeds such as unconditional positive regard, acceptance, respect and dignity for the benefit and advancement of individuals and humankind as a whole. The global nature of the phenomenon of spirituality is also shown by using examples of people who demonstrate compassion and communion with other human beings, in other countries in times of suffering, war and disaster. Compassion and empathy is expressed and experienced for victims of earthquakes that happen miles from home and far removed from personal or religious beliefs. Yet at such times we are all connected in the tapestry of life by our own human spirituality and earthiness. Abstract themes like compassion and justice are treated in the text within the context of spirituality. The author argues that being just and fair means that all patients have the right to achieve spiritual healing regardless of their belief systems, culture or creed. The works of some spiritual philosophers are used to reflect on this integral aspect of human caregiving. Historical symbols of spirituality are examined. The need for nurses to explore and reflect on the paradoxical concepts involved in their own spirituality is highlighted. Nurses are the essential providers of care and, therefore, the paper argues, guardians of that essential humanity that ensures that patients never become less than full human beings, whatever their condition, faith, culture or belief, or whoever they may be. The author contends that this responsibility is uniquely essential to being a nurse. PMID- 9416109 TI - A Baby-Friendly Hospital initiative in northern China. AB - In the People's Republic of China, an approach has been adopted to ensure that all children born as a result of the one-child policy are healthy. The World Health Organization/United Nations Children's Fund (WHO/UNICEF) has encouraged the Chinese Government to embrace this philosophy. Part of this approach has resulted in an attempt to increase the breastfeeding rates. This report describes a visit by two nursing faculty members of a Canadian university to an urban hospital maternity unit in the northeast of China, where the staff were attempting to incorporate this philosophy into their maternity care with the assistance of nursing faculty members from the local university. The method chosen to integrate this philosophy was the Baby-Friendly Hospital Initiative initiated by WHO/UNICEF. PMID- 9416111 TI - Changing health patterns: an enduring puzzle. PMID- 9416110 TI - The human condition of the professional: discretion and accountability. AB - This article takes issue with procedural reductionism, which is the inclination to reduce all matters of judgement and responsibility to the following of some procedure or rule. Two scenarios provide content for a discussion of professional discretion in the context of accountability. The author shows that in professional life there will always be situations that stand beyond the rules of procedures and require the unique judgement of the professional at the time. While this judgement may be determined by the facts available in a situation of uncertainty, it cannot be reduced to the facts (including facts about rules and procedures). The moral judgement will still have an essential indeterminancy about it. PMID- 9416112 TI - Persistence in health patterns: the mystery of being human. PMID- 9416113 TI - Questioning evidence-based practice for nursing. PMID- 9416114 TI - Knowledge and evidence: do they change patterns of health? PMID- 9416115 TI - The paranormal and nursing. PMID- 9416116 TI - Peplau's theory of interpersonal relations. AB - Interpersonal competencies of nurses are key to assisting patients in the work necessary for regaining health and well-being. Peplau's theory of interpersonal relations is detailed, and examples are given of the three phases which occur in developing nurse-patient relationships, along with associated challenges. PMID- 9416117 TI - Peplau's theory in practice. AB - Peplau's theory of interpersonal relations provides a useful framework for investigating clinical phenomena and guiding nurses' actions. The author describes clinical application of selected concepts from Peplau's theory which supports this assumption. While the case data are encouraging, it is suggested that there is a need to test the clinical effectiveness of Peplau's concepts by utilizing experimental research designs. PMID- 9416118 TI - Transforming research and practice with the human becoming theory. AB - This article focuses on the human becoming theory as a guide to the transformation of research and practice. New knowledge gained from human becoming research guides the nurse in practice. Different ways of being in true presence in living the human becoming theory are discussed. PMID- 9416119 TI - Living the art of the human becoming theory. AB - The author shows through examples how quality of life is enhanced when the nurse lives the art of the human becoming theory through composing and playing music with persons and families. Also demonstrated is Parse's practice methodology as lived through music. PMID- 9416120 TI - King's theory of goal attainment in practice. AB - King's conceptual system provides a comprehensive view of three dynamic interacting systems--personal, interpersonal, and social. Her theory of goal attainment has been used as the basis for practice, education, research, and administration, examples of which are presented here. PMID- 9416121 TI - Use of culture care theory with Anglo- and African American elders in a long-term care setting. AB - The purpose of this study was to discover the care expressions, practices, and patterns of elderly Anglo- and African American elders. The domain of inquiry was the cultural care of elderly residents within the environmental context of a long term care institution. The ethnonursing qualitative research method was used to conduct the study which was conceptualized within Leininger's theory of culture care diversity and universality. Four major themes were discovered: (a) Residents expressed and lived generic care to maintain their preadmission lifeways; (b) The nursing staff provided aspects of professional care to support satisfying lifeways for residents; (c) Institutional care patterns and expressions were viewed as a continuing life experience but with major differences between the apartment section and nursing home units; and (d) An institutional culture of the retirement home was discovered which reflected unique lifeways and shared care and health expressions and practices. These themes substantiated the culture care theory and revealed new modes of care for the elderly in an institutional setting. PMID- 9416122 TI - Heart health: treatment. AB - Coronary heart disease (CHD) is the single most common cause of death in England and it has been estimated that about 26 per cent of all deaths each year are caused by CHD (Department of Health 1993). In light of this, it is important that all nurses understand the current investigations and treatment available for people with CHD. PMID- 9416123 TI - The Pew Health Professions Commission reports. PMID- 9416124 TI - Delegation: consultation & education for the professional nurse. PMID- 9416125 TI - Advanced practice nurses: key to the future of health care. PMID- 9416126 TI - The evolution of nursing's social policy statement ... setting new directions for nursing's future. PMID- 9416127 TI - Fundamental principles of ethical conduct in scientific research. AB - A review of historical events pointed to the necessity for ethical conduct in scientific research. The principles of respect for persons, beneficence and justice as identified in the Belmont Report provide valuable guidance for systematic inquiry that adhers to values, standards, and obligations of all disciplines. It is incumbent upon nurses to incorporate these fundamental principles into their research endeavors--whether nurses are in the role of scientific investigator, research participant, or consumer of research findings. PMID- 9416129 TI - Paps getting a bad rap. PMID- 9416130 TI - New perspectives on making a difference. PMID- 9416128 TI - New progress in localizing epilepsy FOCI. PMID- 9416131 TI - NCQA/HEDIS/QARI. What is this alphabet soup all about? PMID- 9416132 TI - Direct acute patient care for seventeen years and "I still want to come to work"! PMID- 9416133 TI - Changes in nursing regulation. PMID- 9416134 TI - Career options: the increasing demand for courtroom nurses. PMID- 9416135 TI - Your interests at the state capitol. A lobbyist! PMID- 9416136 TI - Dollars and cents of APN practice. PMID- 9416137 TI - Novice's account of psychiatric home care nursing. PMID- 9416138 TI - Oklahoma SAFE KIDS coalition. PMID- 9416139 TI - Oklahoma Associate Degree Nursing Educators' position statement on the associate degree. PMID- 9416140 TI - Pregnancy intention and physical violence. PMID- 9416141 TI - My first year. PMID- 9416142 TI - Shoes. PMID- 9416144 TI - Continuing education in nursing--today and into the next century. PMID- 9416143 TI - Get set, get ready, & go, go, go--for the 21st century! PMID- 9416145 TI - Burned out, or burned up? PMID- 9416147 TI - Nurse practitioners of Oregon. PMID- 9416146 TI - Picture of health. PMID- 9416148 TI - ONA remains vocal on multi-state licensure. PMID- 9416149 TI - Multi-state licensure: progressive policy or professional risk? PMID- 9416150 TI - Unit reps are key to success. PMID- 9416152 TI - Wound care: a profile of CNS practice. PMID- 9416151 TI - ONA helps OHD launch statewide latex study. PMID- 9416153 TI - The Nutritional Screening Initiative: meeting the nutritional needs of elders. AB - Nutritional factors are associated with at least 5 of the 10 leading causes of death in the United States. Older persons are especially vulnerable for nutritional problems, usually resulting from inappropriate (usually inadequate) intake of nutrients or increased need for nutrients. Nutritional assessment and interventions are sometimes overlooked in the focus on more complicated medical care. Yet there are many ways in which nurses can intervene to improve patients' nutritional status. One resource for organizing nutritional assessment and interventions is the Nutrition Screening Initiative. This multidisciplinary initiative provides useful guidelines and tools for assessing nutritional status and for planning the most appropriate interventions. PMID- 9416154 TI - Recognizing spiritual needs of orthopaedic patients. AB - Orthopaedic nurses often are well-educated in dealing with patients' physical and psychologic needs but lack education in caring for the spiritual needs of man. Nurses must realize they, themselves, have spiritual needs and must invest in clarifying their own values and beliefs as well as their patients. To perform a complete spiritual assessment, nurses need to become familiar with the concept of spirituality and what it means in the care of patients. Providing spiritual care is individualized and often complex. The nursing process enables the nurse to plan patient care. Providing spiritual care is a challenge orthopaedic nurses must recognize and assume responsibility for. PMID- 9416155 TI - Historical perspectives on orthopaedic nursing research in Orthopaedic Nursing from 1982 to 1995. AB - PURPOSE: To describe a systematic review of research studies published from 1982 through 1995 in Orthopaedic Nursing since its first issue in 1982. DESIGN: A descriptive design was used to systematically examine and classify the research studies published in Orthopaedic Nursing. SAMPLE: Forty-nine research studies made up the sample. METHOD: A descriptive historical research method was used to categorize the studies according to (1) topic, (2) research design, (3) sampling method, (4) sample size, (5) educational preparation of the author(s), and (6) funding status of the study. MAIN RESEARCH CLASSIFICATION: Nursing research Orthopaedic Nursing. FINDINGS: The number of research studies published in Orthopaedic Nursing was higher from 1990 to 1995. Most studies consisted of adult clients in acute care settings. There was increased attention to nursing interventions, psychosocial needs, and professional development (1990 to 1995). Pain was the most frequently addressed topic. Nonexperimental descriptive and retrospective designs were used most frequently. Nonprobability sampling was more common. Numbers of studies acknowledging funding increased from one study (1982 to 1989), to 10 studies (1990 to 1995). CONCLUSION: Overall, the number of research studies published in Orthopaedic Nursing has increased over the years and the studies have become more sophisticated. The clinical setting provides a wealth of topics for studies on nursing interventions and outcomes. IMPLICATIONS FOR NURSING RESEARCH: The final step of the research process, dissemination of research findings is integral to the success of the research process. As nursing continues to grow as a profession, the dissemination of research findings to the practicing clinician provides research-based information that can guide and improve clinical practice on a day-to-day basis. Nurses need to take advantage of the opportunities for conducting and using research. PMID- 9416156 TI - Nursing care of sports-related injuries. PMID- 9416157 TI - Shoulder immobilization devices. AB - Currently a myriad of devices are available for immobilization of the injured or postsurgical upper extremity. Some of these devices are straightforward and easily used, but some are more complicated and require more familiarity for their successful application. However, even simple devices have the potential for misapplication and thus prevent their benefit to the patient. This article is the third in a 3-part series. The goals of the series are (1) to present and review several devices on the market used by shoulder surgeons to immobilize the upper extremity, and (2) to discuss proper application and precautions of their use. It is intended that this series will benefit nurses, therapists, and trainers involved in the use of these devices. PMID- 9416158 TI - Epidural pain management for the pediatric spinal fusion patient. AB - Pain management immediately following a spinal fusion is an important issue for patients and nurses. This article describes the procedure, details nursing considerations for epidural pain management following spinal fusion surgery, and includes data from our institution's 2 years of experience with this type of pain management. With proper nursing education, anesthesia management, and orthopaedic surgeon support, epidural analgesia following spinal fusion surgery is a viable alternative to conventional methods. PMID- 9416159 TI - The problems with "heroic measures": the case of Mr. X. PMID- 9416160 TI - Calcific tendinitis of the shoulder. AB - Calcific deposits in tendinous tissue are common to middle-aged patients. The condition usually resolves spontaneously, but refractory cases can be expedited by needle barbotage or surgery. PMID- 9416161 TI - Pharmacologic management of obesity. AB - Obesity is a major health problem in the United States. It is not only a problem in itself, but it is a predisposing factor for many chronic illnesses. Pharmaceutical houses have devoted much research in the development of drugs to combat the problem. While diet and exercise continue to be the first line of defense, many diet drugs are available on the market to help an individual lose weight. This article discusses some of the theories that appear to explain the development of the overweight condition and provides basic information on the drugs currently in use. PMID- 9416162 TI - Nurses warned about preventing allergic reactions to latex at work. PMID- 9416163 TI - Delegation remains priority practice issue. PMID- 9416164 TI - Boards of nursing recommend mutual recognition model of regulation. PMID- 9416165 TI - Nursing in Guam--challenges and opportunities. PMID- 9416166 TI - ANA hosts immunization meeting between CDC and state nurses' associations. PMID- 9416167 TI - State board continues to deliberate LPNs taking oral orders. PMID- 9416168 TI - Dealing with the panel interview. PMID- 9416169 TI - Entry into practice: a ten year anniversary. PMID- 9416171 TI - "Nurses: the helping hands of disaster". PMID- 9416170 TI - Telenursing--telehealth. PMID- 9416172 TI - Nurse shares personal flood experience. PMID- 9416173 TI - The five hundred year flood. PMID- 9416174 TI - Oncology nurses story. PMID- 9416175 TI - A forty-eight hour day for Valley Memorial Homes in Grand Forks. PMID- 9416177 TI - Queensland Health--for sale? PMID- 9416176 TI - Nurses incorporate for testing center. PMID- 9416178 TI - Health training for women in Vietnam. PMID- 9416179 TI - Would you? ... Could you? ... Put your mother in a nursing home? PMID- 9416180 TI - Nursing Career Structure. AB - This is the first in a series of articles on the Nursing Career Structure. The Nursing Career Structure was achieved in Queensland from a decision of the Australian Industrial Relations Commission (AIRC) in August 1990 and subsequent Queensland Industrial Relations Commission (QIRC) decision of December 1990. The QNU believes it is appropriate given the time since these decisions to revisit the career structure with its members. PMID- 9416181 TI - Compulsory closure of wards/departments in Queensland Health facilities during the Christmas/New Year period. PMID- 9416182 TI - Patient nurse dependency. PMID- 9416183 TI - The new Health (Drugs and Poisons) Regulation. PMID- 9416185 TI - Nurses in Profile. Pat Turner, Occupational Health Adviser, Shell, Qld. Mike Power Community Health Nurse. PMID- 9416184 TI - Enterprise Bargaining in the public sector. PMID- 9416186 TI - The road to Jerusalem. PMID- 9416187 TI - Doris' day out. AB - This article describes an experience that occurred when, as a student nurse on an aged care placement, I saw an opportunity to motivate and empower a client whom I felt would be able, with an individualised approach, to take a step toward making a significant improvement in her quality of life. I believe we, as nurse practitioners, can influence our client's motivation levels in many ways, the most prevalent and effective being through realistic goal setting, altruism and empowerment. PMID- 9416188 TI - The role of exercise in rehabilitation for patients with end-stage renal disease. AB - End-stage renal disease (ESRD) is a major health problem in the United States. Many patients with ESRD experience a decline in physical functioning as a result of the disease process and its associated sequelae. Cardiovascular changes, anemia, and skeletal muscle weakness contribute significantly to this decreased capacity, leading, in many instances, to a primarily sedentary lifestyle. Studies conducted on the effectiveness of exercise training for patients with ESRD reveal numerous physiological and psychological benefits, particularly when training is continued for several months. However, the number of structured exercise programs available as part of a rehabilitation program for ESRD patients is limited. This article provides an overview of the role of exercise for patients with renal disease, and a case study illustrates how nurses, in collaboration with the interdisciplinary team, can be effective in preventing continued deconditioning and in maintaining a more positive outlook in patients with ESRD. PMID- 9416189 TI - Activity-based resource allocation: a system for predicting nursing costs. AB - As hospital-based managers are being confronted with changing patterns of reimbursement, ranging from revenue generating to cost management, it is imperative that hospitals know the exact nursing costs associated with the actual care delivered to specific patients. Nursing care has traditionally been bundled into the room rate for patients. This approach is extremely limiting when facilities are negotiating per diem rates and capitated rate contracts. At Braintree Hospital Rehabilitation Network, the nursing department has developed and implemented an activity-based management system to determine the actual cost of nursing care provided to each patient. This approach, which differentiates nursing costs accurately by diagnostic group and by intensity of nursing care, has contributed to the hospital's success in negotiating individual patient contracts with insurers in the managed care environment that increasingly focuses on costs and outcomes. Another result has been to enhance the accuracy of the network's cost accounting system. PMID- 9416190 TI - A study comparing sterile and nonsterile urethral catheterization in patients with spinal cord injury. AB - This study was designed to determine the effect of sterile and nonsterile intermittent catheterization on the incidence of urinary tract infection (UTI) in patients after spinal cord injury. The study included 29 patients with neurogenic bladder dysfunction treated with intermittent catheterization. One group of 14 patients was on sterile catheterization; another group of 15 patients was on nonsterile catheterization. On a weekly basis, urine samples were obtained and analyzed. A total of 122 urine samples were analyzed. The patients on sterile catheterization had a 28.6% UTI incidence; the group using a nonsterile catheterization technique had a UTI incidence of 42.4%. The most common urinary pathogen in both groups was E. coli (65%). The cost of antibiotics for patients on the sterile catheterization program was only 43% of the cost of antibiotics for those on the nonsterile program. However, the sterile kits cost 371% of the cost of the catheterization kits for the patients in the nonsterile program, so the total cost of managing neurogenic bladder on the sterile program was 277% of the cost of the nonsterile program. PMID- 9416191 TI - Health promotion among ethnic minorities: the importance of cultural phenomena. AB - Ethnic minority groups have varying health promotion needs, which differ according to six cultural phenomena outlined in the nursing literature: communication, space, social organization, time, environmental control, and biological variations. This article reports on some of the research available in each of these areas in relation to ethnic minorities. If health promotion is to be improved for people in all sectors of the population, it is important for rehabilitation nurses to learn how to assess the needs of ethnic minorities and to respond to those needs with appropriate interventions. PMID- 9416192 TI - Patient literacy levels: a consideration when designing patient education programs. AB - The purposes of this study were to examine (a) the relationship between patients' own reports of the highest grade completed in school and their actual reading level and (b) the relationship between literacy and the level of knowledge about self-care after patients had received education involving written discharge instructions. In addition, the content of the materials was analyzed for its cultural sensitivity. Twenty-six patients who had had either hip- or knee replacement surgery at an inner-city hospital participated in this correlational descriptive study. There was a significant negative relationship between patients' own reports of highest grade completed in school and their actual reading level (r = -.39, p < .05). Rehabilitation nurses should find this study beneficial for developing, assessing, and using written patient education materials appropriate for the reading level of their patient populations. PMID- 9416193 TI - Using a microwave oven to disinfect intermittent-use catheters. PMID- 9416194 TI - Individualized educational programs for rheumatology patients. PMID- 9416195 TI - [Spinal cord lesion. Nursing procedure]. AB - There has been a considerable increase in recent years in the number of persons who suffer a spinal cord lesion, winding up either paraplegics or tetraplegics. Their quality of life depends to a large degree on the attention provided by healthcare professionals, both during their period of rehabilitation as well as later, assisting them to avoid those problems which develop. The authors propose to provide the reader with a grasp of what it means to live with a spinal cord lesion. They will comment on the alterations which develop, detailing what monitoring and care the nurse should offer for each one: state how one can avoid or solve problems related to these lesions, point out what the most frequent complications are and inform what aspects one must consider if a patient has to have surgery. PMID- 9416196 TI - [Silastic catheters: pinpointing the end tip of the catheter by means of electrocardiographic monitoring]. AB - The placement of catheters with a silastic center has been a common procedure in neonatal intensive care units for several years. Nonetheless, this procedure, like many others, bears its risks and complications if not properly carried out. The majority of complications, which are described in medical journals, include arrhythmias, myocardiac perforations, thrombosis, hemorrhage in the pleura, etc., and these are related with the catheter and its possible movement inside the blood vessel where it was originally inserted. The usual exploratory procedure to pinpoint the end tip of the catheter has been an ordinary x-ray, but often this x ray does not allow one to see precisely where the catheter tip is located. This problem is caused by the tiny catheter calibre which does not allow for all the necessary contrast; because of this, it is frequently necessary to administer a radiopaque contrasting sub-stance and then repeat the x-ray in order to ensure that the catheter tip is located exactly where it should be. By means of electrocardiographic monitoring, a three-pronged key with an electrode and a 5.85% sodium chloride solution, it is possible to pinpoint the end tip of the catheter without resorting to an x-ray nor administering a contrasting solution. PMID- 9416197 TI - [Antitetanus vaccination: noncompliance with doses]. AB - The objectives of this article are to estimate the percentage of people who fail to receive their second and third dose of the antitetanus vaccination, find a relation between public health information and the reason for administering the vaccine with the percentage of non-compliers, and to estimate the percentage of patients who fail to receive their third dose after a six month waiting period. This study was designed with a multiple follow-up. This study took place at a Primary Health care center: E.A.P. Barcelona II in Mostoles, Madrid. Patients included all those people who met the requirements for the vaccination; in this study there were 452 persons. STATISTICS AND MAIN RESULTS INCLUDE: The percentage of noncompliance with second and third doses was 59.7% (195% (54.4%-65.0%)) There was a higher percentage of noncompliance in the group which received sanitary information, 61.8% while the group which received basic information registered a 54% rate (p > 0.05). The percentage who failed to continue their doses if first administered for a cut was 60.3% against a 59.0% rate for those caught in doctors check-ups, (p > 0.05). The proportion of noncompliance due to the 6 month waiting period was 46.9% (IC95 (37.8-56%)). CONCLUSION: This study did not demonstrate that greater sanitary information coupled with a cut contributed to a higher compliance to the convention vaccination pattern. We note the percentage of noncompliance for the third dose following a 12 month pattern was 55.3% against 46.9% in the six month pattern; this leads us to plan a further study. One can include that more than half the people do not comply when they should for the administration of the antitetanus vaccine. We feel it is necessary to improve the means through which people remember they need to take their antitetanus vaccine's doses. PMID- 9416198 TI - [Are hospitals changing?]. AB - Thoughts regarding the various changes in the evolution of the health care industry, particularly the idea of product, professional relationships and management models. Commentary regarding the development of organizations like the magnet hospitals in the U.S. leads to the question: the level of nursing is higher than ever but are we changing the hospitals (where are a majority of nurses work) to take advantage of that level of knowledge that nurses have attained? PMID- 9416199 TI - [Extracting pumps for breastfeeding]. AB - Nursing mothers can use a variety of devices to aid them while they are breastfeeding their children. Among these are pumps which extract milk from their breasts. In this article, the general features of different types of extracting pumps available to nursing mothers are described. Nurses will be able to acquire knowledge not only about how these devices work but nurses can learn when their use is indicated, what their exact operating procedures are, as well as their advantages and disadvantages. Nurses shall then be better prepared to advise nursing mothers as to which device is most appropriate for them in any given circumstance. Therefore, nurses can provide proper health care leading to the continuation and success of breastfeeding. PMID- 9416200 TI - [What is menopause? (I) Physical symptoms, hormonal factors, and psychological aspects]. AB - This is the first part of a long article which deals with all aspects of menopause. In this part, the physiological processes which occur during this phase in a woman's life are analyzed. PMID- 9416201 TI - [Long-term indwelling catheters. First national congress]. PMID- 9416202 TI - [Indwelling catheters. Indications and methods of insertion]. PMID- 9416203 TI - [Implantable catheters]. PMID- 9416205 TI - [Manipulations of an implant site. Good practice recommendations]. PMID- 9416206 TI - [Patient education]. PMID- 9416204 TI - [Indwelling catheters. Quality assurance in nursing care. Evaluation of professional practices]. PMID- 9416207 TI - [After-care of patients with indwelling catheters at home]. PMID- 9416208 TI - [Epidemiological surveillance of venous accesses. Medical Center of Forcilles]. PMID- 9416209 TI - [Management of cases with complications]. PMID- 9416210 TI - [Nutritional support and HIV]. PMID- 9416211 TI - [Fatigue in oncology]. PMID- 9416212 TI - [Hematology, mucositis and toothbrushing!]. PMID- 9416213 TI - [An information tool: the CD photos]. PMID- 9416214 TI - [Clinical ethics and nursing practice. Second part]. PMID- 9416215 TI - [Tomorrow's accreditation]. PMID- 9416216 TI - [Education of nursing students]. PMID- 9416218 TI - [All the senses of sense]. PMID- 9416217 TI - [Multiple action neuro-hormonal antagonist: carvedilol]. PMID- 9416219 TI - [Sense of care, care of senses]. PMID- 9416220 TI - [Analysis of practices in the praxis of care]. PMID- 9416221 TI - [Educating students to make sense of their practice]. PMID- 9416222 TI - [Patient advocacy and nursing care project]. PMID- 9416223 TI - [The universe of nursing care]. PMID- 9416224 TI - [Persons in precarious situations. Le Groupe, Afedi Landes-Pyrenees Atlantiques]. PMID- 9416225 TI - [Patient's rights]. PMID- 9416226 TI - [Needs of the hospitalized patient]. PMID- 9416227 TI - [A tool to preserve motivation]. PMID- 9416228 TI - [Inservice training is also collaborating]. PMID- 9416229 TI - [The stakes of professional education]. PMID- 9416230 TI - [An engaged professional]. PMID- 9416231 TI - ["The Rainbow Space orders my life"]. PMID- 9416232 TI - [Day hospital]. PMID- 9416233 TI - [Prescriptions of drugs in the elderly. Prudence!]. PMID- 9416234 TI - [Cataracts in the elderly]. PMID- 9416235 TI - [Malnutrition at home and in institutions]. PMID- 9416236 TI - [Story telling workshop in geriatric psychiatry]. PMID- 9416237 TI - [The body in geriatrics: a psychomotor approach]. PMID- 9416238 TI - [Restrained in the hospital: ethical reflections on a practice that is not always so...]. PMID- 9416239 TI - [Bed-chairs: selection criteria]. PMID- 9416240 TI - [Health care actor: the laundress]. PMID- 9416241 TI - Accountability for HMOs. PMID- 9416242 TI - Nursing Educators. Moving forward. PMID- 9416244 TI - The rock-solid profession. PMID- 9416243 TI - In and out of hospital DNR. PMID- 9416245 TI - To be or not to be a part of the solution. PMID- 9416246 TI - Allocation of scarce resources in surgery. PMID- 9416247 TI - The risks associated with pneumonectomy. PMID- 9416248 TI - Aorta to left atrial fistula. PMID- 9416249 TI - Case 18. Rupture of a splenic artery pseudoaneurysm into a pancreatic pseudocyst. PMID- 9416250 TI - Anticoagulation in spinal surgery. A critical review of the literature. AB - OBJECTIVE: To determine the natural incidence of thromboembolic complications and the effect of thromboprophylaxis associated with elective spinal surgery. DATA SOURCES: A search of the MEDLINE database, using the key words anticoagulation, deep vein thrombosis (DVT) and spine, alone and in different combinations. Individual journals were also searched. Articles investigating the incidence or treatment (or both) of thromboembolism in elective spinal surgery were identified. STUDY SELECTION: Studies describing elective spinal surgery. The type of surgery, days of recumbency, methods of thromboprophylaxis, study design, surveillance methods, rates of DVT and pulmonary embolism (PE), and type and rates of complications of thromboprophylaxis were determined. DATA EXTRACTION: Single observer. DATA SYNTHESIS: Only 15 studies were found. Most were of poor statistical quality: 5 were level IV quality (nonrandomized, historic controls), 8 were level V quality (no controls, cases series), 1 was a level III study (nonrandomized, contemporaneous controls) and 1 was a level II study (small randomized study, moderate to high risk of error). The raw incidence of thromboembolic complications derived from these studies was 7.1% (14.1%) (mean [and SD]). However, because of the poor quality of these studies, this figure is suspect. CONCLUSIONS: The true incidence of thromboembolic complication in spinal surgery remains unknown. Recommendations for thromboprophylaxis cannot be made from the findings of these studies. There is a need for a well-designed, randomized controlled study to define the efficacy of thromboprophylaxis in elective spinal surgery. PMID- 9416251 TI - The ethical allocation of scarce resources in surgery: implants and cost. AB - This paper is a discussion of the factors involved in instituting a bulk purchasing program for surgical supplies. An improved understanding of the surgical procedure of joint arthroplasty must relate to the variability in surgical methods that achieve patient outcomes. An understanding of the outcomes in relation to the expected duration of the success of an implant and the high costs associated with a revision earlier than expected must be factored into the budget and costs of implants. The ethical implications of choosing one implant over another are considered. A more uniform outcome assessment with respect to surgical activities is needed and potential savings related to other operating room costs must be examined. Optimizing the implant to patient requirements is the goal within the framework of current fiscal constraints. PMID- 9416252 TI - Comparative study of continuous extrapleural intercostal nerve block and lumbar epidural morphine in post-thoracotomy pain. AB - OBJECTIVES: To compare the efficacy of continuous extrapleural intercostal nerve block with bupivacaine 0.5% in 1:200,000 epinephrine and continuous lumbar epidural block with morphine in controlling post-thoracotomy pain and to measure serum bupivacaine concentrations during extrapleural infusion. DESIGN: A prospective, randomized, controlled trial. SETTING: St. Joseph's Hospital, Hamilton, Ont., a tertiary care teaching centre. PATIENTS: Sixty-one patients booked for elective thoracotomy were randomized by scaled envelope to two groups. INTERVENTIONS: Group A received a continuous extrapleural intercostal nerve block with bupivacaine 0.5% in 1:200,000 epinephrine as a bolus of 0.3 mL/kg followed by an infusion of 0.1 mL/kg every hour for 72 hours. Group B received a continuous lumbar epidural block with morphine as a bolus of 70 g/kg followed by an infusion of 7 g/kg every hour for 72 hours. MAIN OUTCOME MEASURES: Pain was assessed by a linear visual analogue scale (VAS) pain score. The cumulative amount of "rescue" intravenous morphine used, and serum bupivacaine concentrations were measured as secondary outcomes. RESULTS: Pain control was the same in both groups as assessed by linear VAS score (p = 0.33). The cumulative dose of intravenous morphine for supplemental analgesia was statistically significant between the groups: group A patients used more morphine than group B (p < 0.05). Accumulation of serum bupivacaine was present with no clinical toxicity. CONCLUSIONS: There is no significant difference in the degree of post thoracotomy pain control measured by the VAS score when analgesia is provided by continuous extrapleural intercostal nerve block with bupivacaine 0.5% in 1:200,000 epinephrine or lumbar epidural block with morphine. Larger amounts of rescue analgesia were used by patients in the continuous extrapleural group with bupivacaine than those in the continuous lumbar epidural block with morphine. Serum bupivacaine concentrations rise without clinical toxicity. PMID- 9416253 TI - Perioperative mortality after pneumonectomy: analysis of risk factors and review of the literature. AB - OBJECTIVE: To determine risk factors for perioperative death associated with pneumonectomy. DESIGN: A retrospective case-control study in which a perioperative death group was compared with a survivor group, and a review of the English literature on the subject. SETTING: The Montreal General Hospital, a tertiary-care teaching institution. PATIENTS AND INTERVENTION: Ninety-two consecutive patients who underwent pneumonectomy between April 1989 and 1994. MAIN OUTCOME MEASURES: The effects of age, sex, smoking history, tumour size, type and stage, pulmonary function, cardiovascular risks, comorbidity, preoperative blood values and volume of fluids administered perioperatively. Values from the literature were reported for comparison. RESULTS: The perioperative death rate was 10.9%. Selection bias and in-hospital values reported in the literature have underestimated the death rate, with actual rates ranging from 7% to 11%. Age (odds ratio 2.48, p = 0.04), the presence of 1 or more comorbid diseases (odds ratio 7.92, p = 0.05) and amount of fluids given in the first 12 hours postoperatively (odds ratio 2.21, p = 0.06) were found to be significant risk factors for death. Multivariate logistic regression demonstrated that the volume of fluids given remains an independent risk factor whereas age and comorbid disease are dependent variables. CONCLUSIONS: The results were consistent with previously reported death rates and risk factors. Patient age and concomitant disease are not modifiable risk factors, but perioperative fluid administration and other means to prevent postpneumonectomy pulmonary edema may reduce the perioperative death rate. PMID- 9416254 TI - Is prophylactic resection valid as an indication for elective surgery in diverticular disease? AB - OBJECTIVE: To determine whether interval resection in asymptomatic patients after 1 or 2 episodes of acute diverticulitis (prophylactic resection) is justified as a means of preventing late inflammatory complications of diverticular disease. DESIGN: A retrospective analysis. SETTING: A university-affiliated tertiary care hospital. PATIENTS: Those requiring hospitalization from 1987 to 1995 for treatment of acquired diverticular disease of the colon. Twenty-eight patients underwent elective resection and 154 were treated for inflammatory complications (perforation, fistula, complete large-bowel obstruction). INTERVENTIONS: Standard surgical management for diverticular disease, but only 3 prophylactic resections were undertaken during this period. OUTCOME MEASURES: Type of operation, stoma creation and closure, hospital death. In those treated for complicated disease, the effects on outcome of all previous outpatient treatment and hospitalizations. RESULTS: Only 10% of those presenting with complications had been treated conservatively for acute diverticulitis and only 5% had been hospitalized for this reason. CONCLUSIONS: Prophylactic resection is unlikely to prevent late major complications of diverticular disease; therefore, as an elective indication for surgery in this disease its use is questionable. PMID- 9416255 TI - Benign skull lesions. AB - OBJECTIVE: To document the epidemiologic and clinical features of benign skull lesions. DESIGN: A case series. SETTING: St. Michael's Hospital, a tertiary care facility affiliated with the University of Toronto. PATIENTS: Thirty-one patients who had a neurosurgical consultation and were discharged from hospital after excision of a benign skull lesion during a 10-year period. MAIN OUTCOME MEASURES: Patient demographics, clinical signs and symptoms, radiographic and pathological tumour characteristics, surgical procedure, length of hospital stay, outcome and follow-up. RESULTS: The 31 patients (6 men, 25 women) had 32 lesions excised. The mean age of the patients was 41.9 years. Osteomas accounted for 63% of the tumours. The most frequent location was the parietal bone. Neurologic symptoms were absent in the majority of calvarial tumours. Useful diagnostic studies included plain skull radiography and computed tomography. Nuclear bone scanning was done in 7 patients. All patients underwent craniectomy, with cranioplasty in most cases. Three patients had new neurologic symptoms postoperatively and 1 patient had incomplete resolution of symptoms. CONCLUSIONS: Benign skull lesions are infrequent, but they require neurosurgical intervention. When necessary, surgical excision can serve to confirm the diagnosis, improve cosmesis and retard the progression of neurologic dysfunction. Of primary importance is the recognition of such lesions by primary care physicians and referral to the surgeon so that an appropriate treatment plan can be made. PMID- 9416256 TI - Bowel preparation with polyethylene glycol electrolyte lavage solution is potentially hazardous in patients with carcinoma of the cardia: a case report. AB - Polyethylene glycol (PEG) electrolyte solution can induce potentially fatal complications when used as a lavage in preoperative bowel preparation. A 60-year old man with carcinoma of the esophagogastric junction had a Mallory-Weiss tear after bowel preparation with PEG. He was successfully resuscitated, first with balanced salt solution then by transfusion. The literature on the subject is reviewed and recommendations are made, which include consideration of alternative methods of bowel preparation (e.g., sodium phosphate solution) in patients with esophageal obstruction. PMID- 9416257 TI - Malignant giant cell tumour of the distal femur treated by excision, allografting and ligamentous reconstruction: an 18-year follow-up. AB - Extensive osteoarticular allografts have been used for knee reconstruction, but because of their composite nature and the technical difficulty of the procedure, complication and failure rates have been high. There are few records of long-term results in the literature. In this report, a 19-year-old man with a large aggressive giant cell tumour of the left distal femur was treated in 1976 by en bloc resection, massive femoral allografting and ligamentous reconstruction. Follow-up after 18 years showed no recurrence of the tumour, excellent incorporation of the graft and good knee function, which allowed the patient to work 9 hours a day on his feet without pain. PMID- 9416258 TI - Ectopic gallbladder revisited, laparoscopically: a case report. AB - A case of the rare congenital anomaly ectopic gallbladder is presented. A 16-year old girl suffered attacks of epigastric pain unrelated to eating. On abdominal ultrasonography, the gallbladder could not be found in its usual position. Endoscopic retrograde cholangiography demonstrated the gallbladder on the left side of the common duct and the cystic duct arising from the right hepatic duct. Laparoscopic cholecystectomy was done without complication. This appears to be the first reported case of laparoscopic removal of an ectopic gallbladder. The importance of preoperative cholangiography is emphasized for accurate diagnosis and preoperative location of the gallbladder. PMID- 9416259 TI - Superior mesenteric vein thrombosis after the Whipple procedure: an aggressive, combined treatment approach. AB - It is now recognized that occlusion of the mesenteric veins not only may complicate a number of disease processes but may occur as a life-threatening complication after abdominal surgery. A 32-year-old woman had mesenteric venous thrombosis after resection of a duodenal inflammatory pseudotumour by pancreatoduodenectomy. She recovered fully after treatment, which consisted of thrombectomy, flushing with urokinase and intravenous administration of heparin. Papaverine infused for 4 days substantially improved bowel viability. Current concepts in mesenteric vein occlusion and the principles of clinical management are reviewed. PMID- 9416260 TI - Bowel obstruction in a pregnant patient with ileal pouch-anal anastomosis. AB - Bowel obstruction is a rare but serious complication of pregnancy for both the mother and the developing fetus. This report describes the case of 17-year-old girl with ileal pouch-anal anastomosis (IPAA). She presented at 36 weeks' gestation with a complete small-bowel obstruction. Because conservative management was unsuccessful, labour was induced to relieve the obstruction or simplify surgery. Soon after spontaneous vaginal delivery she began to pass copious amounts of flatus and stool. The bowel obstruction resolved within hours. This report illustrates how IPAA alters the anatomy of the gastrointestinal tract, placing the ileal pouch at risk from compressive obstruction by the gravid uterus. Induction of labour in a near-term fetus is a reasonable initial method of management in such women. PMID- 9416261 TI - Glutamate uptake and Na,K-ATPase activity in rat astrocyte cultures exposed to ischemia. AB - In this study we demonstrate the stimulation of both glutamate uptake and Na,K ATPase activity in rat astrocyte cultures in response to a sublethal ischemic insult in vitro. To measure sodium pump activity and glutamate uptake, 3H glutamate and 86Rb were simultaneously added to the cultures in the presence or absence of 2 mM ouabain. Na,K-ATPase activity was defined as ouabain-sensitive 86Rb uptake. Cell death was assessed by exclusion of the vital dye, calcein-AM from cells. Concomitant transient increases (2-3 fold above control levels) in both Na,K-ATPase and glutamate transporter activities were observed in astrocytes after 2-4 hours of ischemia. By contrast, 24 hours of ischemia caused a profound loss of both activities which paralleled significant cell death. The addition of 5 mM glucose to the cells after 4 hours of ischemia prevented the loss of sodium pump activity and glutamate uptake, and rescued astrocytes from the lethal effects of 24 hours of ischemia. PMID- 9416262 TI - The role of calcium ion in anoxia/reoxygenation damage of cultured brain capillary endothelial cells. AB - Capillary endothelial cells are critical targets in both ischemia and reperfusion of the brain. Arachidonic acids and oxygen free radicals have been shown to cause disruption of blood-brain barrier (BBB) by destruction of capillary endothelial cell membrane. However, the exact mechanism of BBB breakdown by cerebral ischemia/reperfusion remains undetermined. The aim of the present study is to clarify the mechanism of intracellular calcium ion ([Ca2+]i) change in brain capillary endothelial cells under anoxia/reoxygenation. Brains capillary endothelial cells were isolated from ten male Sprague-Dawley rats by a two step enzymatic process. [Ca2+]i was measured by means of a confocal laser scanning microscope using Indo 1-A/M as a calcium indicator. The endothelial cells were subjected to anoxia and reoxygenization under different conditions. [Ca2+]i increased gradually during anoxia and slightly decreased after reoxygenation. Indomethacin and SOD suppressed the elevation of [Ca2+]i during anoxia. NG-nitro L-arginine methyl ester and catalase moderately suppressed the elevation, however nifedipine did not suppress it at all. In this model, rapid [Ca2+]i change was not observed during the reoxygenation phase. The results indicate that the anoxia induced elevation of [Ca2+]i in the brain capillary endothelial cells depends on superoxide and peroxynitrite generation. PMID- 9416263 TI - Hypoxia modulates free radical formation in brain microvascular endothelium. AB - Although free radical species (ROS; i.e., .O2-. .OH.H2O2) among other mediators, may be involved in altering the blood-brain barrier (BBB), little is known about the endogenous ability of cerebromicrovascular endothelium to generate ROS. This study examines the capacity of rat endothelial cells (RBEC) to produce ROS in normoxia and hypoxia/reoxygenation. Cultured RBEC were exposed to an oxygen depleted atmosphere (containing 95% N2 and 5% CO2) for 4 hr at 37 degrees C and air (10 min) at room temperature to simulate "ischemia/reperfusion". Nitroblue tetrazolium (NBT) reduction [formation of nitroblue formazan (NBF)] served as a marker for the production of ROS. The release of lactate dehydrogenase (LDH) and [3H]arachidonic acid (AA) was used to assess cellular integrity. RBEC exposed to hypoxia/reoxygenation produced up to 59% greater NBF formation than controls without affecting the LDH or AA release. The production of ROS was calcium dependent and not affected by AA or its metabolites. The findings indicate that the RBEC can produce superoxide dismutase (SOD)-inhibitable ROS which are augmented by hypoxia/reoxygenation. It is suggested that in vivo cerebromicrovascular endothelium may contribute to the formation of ROS and play a role in ischemic brain edema. PMID- 9416264 TI - Increase in surface expression of ICAM-1, VCAM-1 and E-selectin in human cerebromicrovascular endothelial cells subjected to ischemia-like insults. AB - Secondary ischemic brain injury has been shown to develop as a consequence of inflammation and vasogenic brain edema. In this study we show that inflammatory cytokines and simulated in vitro ischemia stimulate the surface expression of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and endothelial-leukocyte adhesion molecule-1 (E-selectin) in human cerebromicrovascular endothelial cells (HCEC) in culture. The levels of all three adhesion molecules were dramatically (3 to 10-fold) up-regulated by 4-24 hour exposure to the inflammatory cytokines. IL-1 beta (10-200 u/ml) or TNF alpha (50 200 u/ml), and by a 4 hour exposure to "simulated" in vitro ischemia, as determined by immunocytochemistry and ELISA. Following 24 hours of subsequent reperfusion, the expression of ICAM-1 and VCAM-1 was maintained at ischemia induced levels, whereas E-selectin was no longer detectable. Both the cytokine- and ischemia-induced up-regulation of adhesion molecules were completely abolished by the transcriptional inhibitor, actinomycin D (10 micrograms/ml), and inhibited by the cycloxygenase (COX) inhibitor, indomethacin (300 microM). These findings implicate HCEC in the processes of leukocyte adhesion and recruitment in the brain during stroke in vivo. PMID- 9416265 TI - Assessment of the relationship between ischemic damage and brain swelling in frozen brain slices. AB - The purpose of the study was to verify a method for the measurement of both cerebral infarction and brain swelling in frozen brain slices for histology. The animals were divided into two groups sham-operated control (n = 10) and focal cerebral ischemia group (n = 10). Focal cerebral ischemia was produced by permanent occlusion of the left middle cerebral artery. The rats were sacrificed 24 hours postocclusion. The brain was divided into two through the corpus callosum. Each hemisphere was weighted and frozen. Cerebral infarction and brain swelling were each assessed at 8 predetermined coronal planes. The volume of brain swelling was obtained by subtracting the total volume of nonischemic hemisphere from the total volume of the ischemic one. In the MCA occlusion group, brain infarction and the differences of hemispheric volume and weight between the right and left hemispheres were consistently observed, whereas sham-operated rats demonstrated no brain infarction or significant differences between two hemispheres. There were good corelationships not only between the volumes of brain edema and infarction (p < 0.05) but between the volume of brain edema and the difference in weight (p < 0.01) also. The results indicate that the measurement of the volume of ischemic brain edema in frozen brain slices may be useful in elusidating relationship with ischemic brain damage. PMID- 9416266 TI - Brain edema associated with progressive selective neuronal death or impending infarction in the cerebral cortex. AB - This study examined the temporal profile of brain edema in the cerebral cortex associated with selective neuronal death or focal infarction after repeated ischemia at an intensity of ischemic insult just under and above the threshold level to induce infarction. The left carotid artery of adult gerbils was twice occluded for 10 min each time with a 5-hr interval between the blockages. In this model, focal infarction developed in coronal sections examined at the chiasmatic level (face A), whereas only selective neuronal death without infarction was found in the coronal section observed at the infundibular level (face B). In each animal, Evans blue (2%) was intravenously injected 1 hr prior to sacrifice as an indicator of blood-brain barrier (BBB) disruption. Brain edema was assessed by gravimetry in samples taken from both faces at 15 min, 5 hr, 12 hr, 24 hr, and 48 hr after the 2nd 10-min ischemia. Evans blue extravasated only in face A, corresponding to focal infarction at 24 and 48 hr after the 2nd 10-min ischemia. The specific gravities of the ischemic cortex of both faces decreased significantly from control at 15 min (P < 0.05) and had recovered by 5 hr after ischemia. By 12 hr, the specific gravities of both faces had again decreased significantly from the control values (P < 0.05), but did not differ significantly from each other. At 24 and 48 hr, the specific gravities of both faces were significantly lower than the control values (P < 0.01), and the specific gravity of face A was markedly lower than that of face B (P < 0.01). We concluded that in face B, where only selective neuronal death without infarction occurred only cytotoxic edema develops, whereas in face A, where infarction progresses, vasogenic edema develops in addition to cytotoxic edema. PMID- 9416267 TI - Blood volume and flow velocity through parenchymal microvessels in ischemic brain edema of rats. AB - Focal cerebral ischemia was produced in rats with left middle cerebral artery occlusion for 24 hours. Regional CBF was measured by the 14C-iodoantipyrine technique. The distribution of red blood cells (RBC) and plasma in cerebral microvessels was determined by radioluminography using 51Cr-RBC and 125I-bovine serum albumin, respectively. The mean transit times of RBC and plasma, blood volume, and hematocrit were calculated. The water content was measured by specific gravity. The blood flow was reduced to 2% of the control value in the central core, where the brain edema was the most severe. The blood volume decreased to 25% and the mean transit times of RBC and plasma increased about tenfold. In the outer periphery, where CBF was reduced to 39% but brain edema was not induced, the blood volume was decreased to 76% while the mean transit time of RBC was increased 2.1-fold, being greater than the increase in the plasma transit time. These findings indicate that focal ischemia has variable effects on the blood volume and flow velocities of RBC and plasma in the parenchymal microvessels depending on the depth of blood flow and edema. A decrease in blood flow is probably related to a reduction in the flow velocities of RBC and plasma in the surrounding ischemic tissue rather than to decreased number of perfused capillaries in the ischemic core. PMID- 9416268 TI - Hyperglycemia and the vascular effects of cerebral ischemia. AB - Hyperglycemia generally enhances cerebral ischemic injury. Most research has focused on the adverse effect of increased lactate production (acidosis) leading to neuronal injury. The effects of hyperglycemia on another possible primary target, the cerebral microvasculature, is examined in this study. Focal cerebral ischemia was achieved by thread occlusion of the middle cerebral artery (MCA). Preischemic hyperglycemia was induced by intra peritoneal administration of 50% of D-glucose solution. In contrast to normoglycemic controls, glucose-injected rats showed a well demarcated pale infarct after 2 or 4 hours of ischemia reflecting a reduction in cerebral plasma volume (CPV) to 73 +/- 9 and 55 +/- 6% of the contralateral hemisphere by 2 and 4 hours respectively. Cerebral blood flow (CBF) measured by laser Doppler flowmetry indicated that after the initial decline in CBF with MCA occlusion, hyperglycemia led to a further progressive reduction during ischemia. On reperfusion, hyperglycemia resulted in poor restoration of CBF, increased occurrence of hemorrhagic infarction (12 of 12) and a large infarct volume. Hyperglycemia induces progressive cerebrovascular changes during ischemia and affects hemodynamic recovery on reperfusion. These changes may contribute to the adverse effects of hyperglycemia in stroke. A reduction in CPV may be a useful indicator of an increased incidence of hemorrhagic infarction after thrombolytic therapy for ischemic stroke. PMID- 9416269 TI - The effect of non-competitive N-methyl-D-aspartate receptor antagonism on cerebral oedema and cerebral infarct size in the aging ischaemic brain. AB - Although vulnerability to ischemic neuronal injury is enhanced with age, the aging brain may be less amenable to neuroprotection as a result of quantitative and qualitative changes in the NMDA receptor. In addition, the elderly may be less tolerant of adverse effects of neuroprotective drugs and this might ultimately limit therapeutic potential in human stroke. However, antagonism of the excitotoxic effects of glutamate by parenteral administration of the non competitive NMDA antagonist magnesium has been well tolerated and has shown to be neuroprotective in young animal models of stroke and head injury. We therefore evaluated the potential of magnesium chloride to reduce ischemic neuronal injury in aged rodents subjected to permanent occlusion of the middle cerebral artery. Treatment with magnesium chloride induced hypotension and hyperglycaemia in both adult and aged rats and did not reduce ischemic neuronal injury or cerebral edema. However, despite these adverse haemodynamic and biochemical effects, which may augment cerebral infarct size, ischemic damage was not exacerbated in the treated groups, suggesting that magnesium has the potential to salvage penumbral neurons and that inotropic support and maintenance of normoglycaemia may permit realisation of its neuroprotective potential. This may have important implications for future clinical stroke trials. PMID- 9416270 TI - Effects of hyperglycemia on cerebral blood flow and edema formation after carotid artery occlusion in Fischer 344 rats. AB - This study examines whether during bilateral carotid artery occlusion in Fischer 344 rats, hyperglycemia induces cerebrovascular changes that enhance brain edema formation. Preischemic hyperglycemia was induced by intraperitoneal administration of D-glucose solution. Laser-Doppler flowmetry, indicated that after the initial decline in blood flow with carotid occlusion (36 +/- 4% of preischemic), hyperglycemic but not control rats showed a further progressive decrease to 19 +/- 2% of preischemic at 120 minutes (p < 0.001). Brain water content was significantly higher in hypercompared to normoglycemic rats after both 2 hours of permanent occlusion (3.86 +/- 0.05 vs. 3.73 +/- 0.03 g/g dry wt.; p < 0.05) and 2 hours of temporary occlusion followed by 1 hour of reperfusion (4.01 +/- 0.08 vs. 3.71 +/- 0.03 g/g dry wt.; p < 0.05). The difference in brain edema formation between normo- and hyperglycemic rats appears to primarily reflect the effects of hyperglycemia on CBF. Cerebral plasma volume (CPV) 2 hours after occlusion was also reduced in hyper-compared to normoglycemic rats (3.9 +/- 0.9 and 7.2 +/- 0.1 microliters/g; p < 0.01). Thus, hyperglycemia in a model of global ischemia induces a reduction in CPV and progressive decline in CBF. In this model, the decline in CBF is of sufficient magnitude to enhance brain injury as evidenced by edema formation. PMID- 9416271 TI - The mechanism of free radical generation in brain capillary endothelial cells after anoxia and reoxygenation. AB - We studied the mechanism of reoxygenation injury of cerebral microvessels in cultured rat brain capillary endothelial cells (BCECs). BCECs were isolated from rat cerebral cortices by a two step enzymatic treatment. The monolayers of BCECs were subjected to anoxia for 20 minutes followed by reoxygenation for 3 hours. Cell damage was assessed by measuring the leakage of intracellular lactic dehydrogenase (LDH). The control group was anoxia/reoxygenated BCECs without any protective reagents. To study the protective effect of free radical scavengers and antioxidants, superoxide dismutase, catalase, deferoxamine, oxypurinol, indomethacin, or NG-nitro-L-arginine methyl ester (L-NAME) was applied during anoxia/reoxygenation. Thus 7 experimental conditions were established. Lactic dehydrogenase (LDH) leaked from reoxygenated BCECs due to cell membrane damage. This leakage was almost totally suppressed by superoxide dismutase, indicating that reoxygenation injury of BCECs is mediated by superoxide generation. The other scavengers and antioxidants partially suppressed LDH leakage. Reduction of Ca2+ in the culture medium from 1.6 mM to 0.016 mM also suppressed LDH leakage. These results indicate that BCECs subjected to anoxia/reoxygenation become potent generators of superoxide anion, which is thought to be responsible for reoxygenation injury. The superoxide generation partially depends on the xanthine oxidase and cyclooxygenase pathways. As L-NAME partially suppressed LDH leakage peroxynitrite may contribute to reoxygenation injury of BCECs. The extracellular Ca2+ concentration also plays a critical role in the reoxygenation injury of BCECs. PMID- 9416272 TI - Mannitol decreases ICP but does not improve brain-tissue pO2 in severely head injured patients with intracranial hypertension. AB - Little is known about the effect of post-traumatic mannitol infusion on cerebral metabolism and oxygenation. The purpose of this study was to investigate the effects of mannitol in comatose patients on PtiO2, PtiCO2 and brain tissue pH using Clark-type electrodes implanted into cerebral white matter. In the neurosurgical intensive care unit PtiO2, PtiCO2, brain tissue pH, arterial blood pressure, intracranial pressure (ICP), cerebral perfusion pressure (CPP) and jugular bulb oxygen saturation (SjvO2) were prospectively studied in eleven patients with severe traumatic brain injury (TBI) during a total of 30 mannitol administrations (125 ml of 20% Mannitol infused over 30 min through a central vein). When the initial ICP before mannitol infusion was below 20 mmHg neither ICP nor any of the other parameters changed significantly during or after mannitol infusion. With a pre-infusion ICP above 20 mmHg a significant effect was seen on ICP (decrease from 23 +/- 1 to 16 +/- 2 mmHg at 60 min) and CPP (increase from 68 +/- 2 to 80 +/- 3 mmHg at 120 min). These effects were not reflected in PtiO2 or SjvO2, which were 29 +/- 4 mmHg and 61 +/- 3%, respectively, at the beginning of mannitol injection and remained unchanged during the observation period. PtiCO2 and brain tissue pH were not affected by mannitol infusion. Future studies should focus on the identification of ICP or CPP thresholds where infusion of mannitol may actually improve O2-supply to the brain. PMID- 9416273 TI - A chronological evaluation of experimental brain infarct by diffusion-mapping and magnetization transfer contrast imaging. AB - BACKGROUND AND PURPOSE: There is a complex system of evolving physiochemical processes in the ischemic brain. The evaluation of this chain of processes is a major challenge of recent stroke studies. Two magnetic resonance imaging techniques: diffusion-weighted (DW) and magnetization transfer contrast (MTC) imaging were introduced in experimental studies and were shown to have sensitivity for different stages of brain infarct. MATERIALS AND METHODS: We used a reproducible middle cerebral artery (MCA) occlusion model in rat to examine infarcts of different time courses. Magnetic resonance T2-weighted (T2). DW, and MTC imaging were performed 3 h, 1 d, 3 d, 5 d, 2 w, 3 w, and 4 w after MCA occlusion. Haematoxylin/eosin (HE) stained sections, which revealed sub regions within infarct lesions, were compared to T2, DW, diffusion-mapping and MTC mapping images. RESULTS: On DW images 3 hours post occlusion lesions were detected as an area with high signal intensity, while T2 imaging does not raise significant contrast of the lesion at this early stage. Three sub regions of the lesion having different ADCs, are discernible on diffusion-mapping images from 1 day to 7 days. The decrease of MTC effect was measured within the infarct lesion from core to marginal zone, from 7 days to 4 weeks. CONCLUSIONS: Diffusion mapping imaging may help to examine brain infarction from 3 hours to 5 days. MTC imaging recognizes infarcts 7 days after the onset. Following this stage, MTC mapping images may provide a quantitative method to assess infarct size. PMID- 9416275 TI - Heterogeneous distribution of early energy failure in experimental focal ischemia of the cat brain. AB - The distribution of succinate dehydrogenase (SDH) activity and the corresponding changes in specific gravity were studied in cats with experimental focal ischemia. Two hours of tandem occlusion of the middle cerebral artery (MCA) and the conunon carotid artery produced a scattered reduction of SDH activity and corresponding brain edema in the cortex. Recirculation ameliorated the SDH reduction and the scattered pattern disappeared, although the brain edema increased further. Four hours of focal ischemia resulted in diffuse reduction of SDH activity in the MCA-perfused area. The scattered area of SDH reduction after 2 hours of focal cerebral ischemia indicates that the ischemic core is multicentric in the early phase, and that these areas fuse together to form a well demarcated infarction, if the blood flow is not reestablished. A short period of cerebral ischemia produces multicentric small infarcts in the cortex, which resemble granular atrophy. PMID- 9416274 TI - Ultrastructure of astrocytes associated with selective neuronal death of cerebral cortex after repeated ischemia. AB - Astrocytic swelling after ischemic insult has been considered a sign of parturbed cell viability. Investigations using cultured astrocytes and C6 glioma cells have revealed that viable astrocytes swell, spatially buffering various metabolites which are increased by the metabolic turmoil following ischemic insults. In the present study, we have studied the temporal profile of ultrastructural changes of astrocytes in the cerebral cortex associated with progressive selective neuronal, death where infarction is not induced. We occluded the left carotid artery of the Mongolian gerbil twice for 10 minutes at a 5 hr interval. In this model, following reperfusion, selective neuronal death progresses in the coronal section cut at the infundibular level. The whole brains of the sham operated control and postischemic animals were fixed by transcardiac perfusion of glutaraldehyde fixatives, at 15 min, 5 and 12 hr after the 2nd 10 min ischemia. Ultrathin sections including the 3rd and 5th cortical layers were prepared from the cut surface at the level of infundibulum. Mild swelling of astrocytic processes and perivascular end-feet was observed in the 15 min group. Glycogen granules were not prominent. In the 5 hr group, we found a few necrotic neurons disseminated in the cortex. All astrocytic cell processes were swollen with increased number of glycogen granules, especially marked in the perivascular end-feet. In the 12 hr group, necrotic neurons increased in number, astrocytic swelling was more extensive, and glycogen granules were evident in astrocytes. No cellular destruction was observed. We conclude: 1. Swelling progresses in astrocytes which however still remain viable and this process is associated with selective progression of neuronal death. 2. Glycogen granules increase in the swollen yet viable astrocytic cell processes. PMID- 9416277 TI - Selective impairments of mitochondrial respiratory chain activity during aging and ischemic brain damage. AB - Cumulative oxidative damage to mitochondrial deoxyribonucleic acid (DNA) with subsequent defects in oxidative phosphorylation may reduce the capacity of the aging brain to cope with metabolic stress. This may contribute to the age related increase in cerebral infarct size that has been documented following permanent middle cerebral artery occlusion (MCAO) in the rat. This hypothesis was evaluated by assessing mitochondrial respiratory chain complex activity in both ischemic and non ischemic brain tissue of adult (10 month) and aged (28 month) male Wistar rats, six hours after occlusion of the left middle cerebral artery. Aging was associated with a significant decline in cerebral mitochondrial function with impairment of the activities of complexes I. II and IV. The individual respiratory chain complexes also exhibited selective vulnerability to a focal cerebral ischemic lesion, with significant impairment of complex I activity in the lesioned hemisphere of both age groups. The age related decline in complex I activity may be important in the enhanced susceptibility of the aging brain to ischemic neuronal damage. PMID- 9416276 TI - Leukocyte-endothelium interactions in global cerebral ischemia. AB - A closed cranial window was implanted in male Mongolian Gerbils to investigate leukocyte-endothelium interactions (LEI) at the brain surface in global cerebral ischemia (GCI) by intravital fluorescence microscopy. Four days after 15 min of bilateral common carotid artery occlusion the ischemic tissue damage was histologically analysed in selectively vulnerable areas of the brain. The frequency of Rhodamine 6G labeled leukocytes rolling along ("roller") and firmly attached ("sticker") to postcapillary endothelium was assessed before and up to 180 min after GCI. As compared to the sham operated control animals induction of LEI was found in animals with GCI, following a steady increase up to a significant level attained at 60 min (rollers) or at 120 min of reperfusion (sticker), respectively (p < 0.05). In animals with cerebral ischemia histological assessment revealed a significant decrease of viable neurons in the CA1-sector of hippocampus (neurons/mm2 +/- SEM: 1308 +/- 71 vs. 829 +/- 106), in parietal neocortex (727 +/- 17 vs. 542 +/- 49), and in striatum (547 +/- 26 vs. 352 +/- 49; p < 0.01), respectively. A significant correlation between the extent of irreversible neuronal damage and the frequency of leukocyte adherence to the endothelium could not be established. Nevertheless a direct correlation between the number of surviving neurons and of rolling leukocytes was observed, which may be suggestive of a protective potential of leukocyte rolling. PMID- 9416278 TI - Regional differences in the cerebrovascular reserve following acute global ischemia in rabbits. AB - The changes of the blood flow velocity in different cerebral arteries under normal and postischemic conditions were investigated in order to evaluate cerebrovascular reserve capacity after the brain ischemia. The experiments were carried out in rabbits. Administration of acetazolamide (Diamox) of 20 mg/kg was performed in all experimental animals and the blood flow velocity in the middle cerebral artery (MCA) and the basilar artery (BA) was measured using the transcranial Doppler sonography. In the control group of animals the intravenous administration of Diamox caused significant increase of the blood flow velocity in the MCA and the BA. In the second group of animals subjected to the acute global ischemia (occlusion of two vessels + hypotension) of 10 min duration prior to the Diamox administration, no increase of the blood flow velocity in the MCA was observed parallel with an increase in the BA. In some animals there was a decrease in the blood flow velocity in MCA. This could be a consequence of stealing phenomenon, as an increase of cerebral blood flow in the posterior fossa compartment, at the cost of the supratentorial circulation. Identification of the areas damaged after the brain ischemia was performed by the 2,3,5 triphenyltetrazoliumchloride (TTC) staining technique. This revealed more pronounced ischemic lesions in the cerebral cortex in comparison to the region of the basal ganglia. PMID- 9416279 TI - Edema formation exacerbates neurological and histological outcomes after focal cerebral ischemia in CuZn-superoxide dismutase gene knockout mutant mice. AB - In a variety of studies. CuZn-superoxide dismutase (CuZn-SOD) has been shown to protect against ischemic brain injury. A possible role for CuZn-SOD-related modulation of neuronal viability has been suggested by the finding that CuZn-SOD inhibits brain edema formation following various kinds of neurological insults. We have evaluated the role of CuZn-SOD on brain edema formation following focal cerebral ischemia in mice bearing a disruption of the CuZn-SOD gene (Sod1). Homozygous mutants (Sod1-/-) had no detectable CuZn-SOD activity and heterozygous mutants (Sod1+/-) showed a 50% decrease compared to wild-type mice. Sod1-/- mice showed a high level of blood-brain barrier (BBB) disruption shortly after 1 hr of middle cerebral artery occlusion and 100% mortality at 24 hr following ischemia. Sod1+/- mice showed a moderate level of BBB disruption and 30% mortality. The Sod1+/- animals had increased infarct volume and brain swelling, accompanying exacerbated neurological deficits at 24 hr following ischemia. These results indicate the important role of superoxide anions in the development of brain edema after focal cerebral ischemia and suggest the possibility that brain edema formation may contribute to the exacerbation of ischemic brain injury and neurological deficits in knockout mutant mice. PMID- 9416280 TI - Therapeutic dose and timing of administration of RNA synthesis inhibitors for preventing cerebral vasospasm after subarachnoid hemorrhage. AB - A RNA synthesis inhibitor, dactinomycin, intended to suppress induction of vasoconstrictor peptide endothelin, prevented cerebral vasospasm almost completely in the dog subarachnoid hemorrhage (SAM) model [8]. Since endothelin receptor antagonists have not shown so potent effect as dactinomycin in animal SAH models, we aimed clinical use of dactinomycin for improvement of final outcomes of severe SAH patients now suffering vasospasm. Before clinical application, we examined therapeutic dose and timing of administration of dactinomycin in animal models. In the dog two-hemorrhage model, dactinomycin treatment (0.01 mg/kg i.v. for 5 days) started at 6 hours after the second blood injection on Day 2 prevented vasospasm, but that started on Day 3 did not. Low dose of dactinomycin (0.003 mg/kg i.v. for 5 days) rather aggravated vasospasm even though the treatment started on Day 0. In the rat vasospasm model, high dose of dactinomycin (0.03 mg/kg i.p. for 3 days) or relatively low dose of doxorubicin (0.6 mg/kg i.p. once) prevented vasospasm even though the treatment started on Day 4. The present study suggests that RNA synthesis inhibitors, such as dactinomycin and doxorubicin, may aufficiently prevent or ameliorate cerebral vasospasm in severe SAH patients. PMID- 9416281 TI - Traumatic brain swelling in head injured patients: brain edema or vascular engorgement? AB - Brain edema and vascular engorgement have been used interchangeably to describe brain swelling associated with severe brain trauma and their relative contribution of these compartments to the swelling process remains controversial. In this report, imaging techniques for measurement of brain water and blood volume have been used to study the relative contribution of blood volume and tissue water to the swelling process in severely brain injured patients. More specifically, magnetic resonance techniques for non-invasive tissue water measures founded on mathematical models and later substantiated in laboratory and clinical studies were used for measure of brain tissue water. These studies were combined with measures of cerebral blood volume utilizing indicator dilution methods. Studies indicated that brain water was increased while blood volume decreased. These studies provide compelling evidence that the major contributor to brain swelling is brain edema and not blood volume. Therapies should now be targeted toward preventing edema development and enhancing edema resolution. PMID- 9416282 TI - Evaluation of homeostatic changes in CSF circulation: in vivo analysis of the effect of neurotransmitter accumulation in the extracellular space following transient global ischemia. AB - Accumulation of potassium and excitatory amino acids (EAA) in the extracellular space (ECS) following ischemia has been well documented. Careful monitoring of these transients is crucial to gain a better understanding of CNS pathophysiology. This study was initiated to determine if CSF concentrations of EAAs reflect those measured in the ECS. Transient global ischemia, 20 minutes in duration, was produced by clamping the left subclavian and innominate arteries combined with hemorrhagic hypotension. The accumulation of glutamate and electrolytes were measured in CSF and the extracellular fluid (ECF) of cerebral cortex. Microdialysis (MD) was utilized to measure the extracellular concentrations while direct sampling of CSF was provided via cannulation of the cisterna magna. Hydrogen clearance and laser doppler methods were used to monitor regional cortical CBF. Our results show that extracellular concentrations of potassium ([K+]ECF) and glutamate significantly increased following the initiation of ischemia (p < 0.05). The extracellular concentration of these substances decreased with the restoration of CBF. In CSF, a similar trend was observed following re-circulation (p < 0.05). However, CSF glutamate levels did not return to pre-ischemic values. PMID- 9416283 TI - Detection of brain atrophy following traumatic brain injury using gravimetric techniques. AB - We hypothesized, that with atrophy, the correlation between water content and specific gravity of brain solids would break down signifying the onset of the atrophic process. The correlation between tissue water content, specific gravity of solids and ventricular size was studied in an impact acceleration model of closed head injury of the rat. Adult Sprague Dawley rats weighing 350 to 375 grams (n = 63) were separated into two groups: Group 1: Sham (n = 21), Group II: Trauma (n = 42). Water content was assessed using both gravimetric method and drying-weighing method at 1 hour, on days 1, 3, 7, 14, 28, and 42 in the trauma group as well as in the control group. Ventricular size was measured in cm2 on the MRI computer console in the coronal section at the coronal suture at the same time points. In the trauma group we found a significant increase (p < 0.01) in water content during the first week except on day 3 and there was a good correlation between the results of water content using both methods (p < 0.001). However, this relationship was poorly correlated after day 14 (p = 0.25). Although the ventricular size was the smallest at 1 hour post trauma, it significantly increased over the next 3 days (p < 0.001). On day 7 and 14 ventricular size decreased to normal size, yet gradually increased and then reached a significantly larger size on 42 days post trauma again (p < 0.01). We may consider, that brain edema following CHI begins immediately following trauma and resolves within 2 weeks. After 14 days degenerative change occurs in the cortex, as detected by specific gravity measurements which signifies the onset of the atrophic process and subsequent post traumatic ventricular dilatation. PMID- 9416284 TI - Altered brain oxygen extraction with hypoxia and hypotension following deep hypothermic circulatory arrest. AB - The utilization of cardiopulmonary bypass in neonates, infants and children often requires the use of deep hypothermia at 18 degrees C with occasional periods of circulatory arrest. Thus, marked physiologic extremes of temperature and perfusion are induced. The safety of these techniques appears to be related to the reduction of metabolism, particularly cerebral metabolism. We studied the effect of deep hypothermic circulatory arrest and cardiopulmonary bypass on brain oxygenation using near-infrared spectroscopy. After hypothermic arrest, brain oxygen extraction during severe hypoxia and severe hypotension is diminished. However, these responses remain intact after cardiopulmonary bypass. Additionally, cardiopulmonary bypass, rather than deep hypothermic circulatory arrest alters the cerebral oxygen response to hypercapnia. The primary goal of studying alteration of brain oxygenation during cardiopulmonary bypass and deep hypothermic circulatory arrest is to improve our understanding of the association between these methods and perturbations in hemodynamics and ventilation, so that effective brain protection strategies can be developed. PMID- 9416285 TI - Posttraumatic edema in the corpus callosum shown by MRI. AB - MRI was performed on 120 patients who sustained closed head injury of varying severity. Patients ranged in age from 4 to 87 years (average, 32 years). All patients had an initial MRI within 28 days (median 12 days) of injury. MRI disclosed areas of abnormal signals in the corpus callosum of 21 (18%) of the 120 patients; 1 (2%) of the 44 patients who sustained mild injuries (GCS > or = 13), 3 (10%) of the 31 moderate injuries (GCS 9-12), and 17 (38%) of the 45 severe injuries (GCS < or = 8) (p < 0.0001). All but 2 of the 21 patients with corpus callosum lesions had other parenchymal lesions that were visualized by MRI. Of these 21 patients, MRI was repeated in 19. In 13 of the 19 patients, repeat MRI scans at 25 to 42 days after injury showed the disappearance of lesions that had on the first MRI shown a high signal on T2-weighted and FLAIR images and a normal signal on T1-weighted images. The MRI findings and time source of the disappearance of the corpus callosum lesions mirrored those of paracontusional edema in the subcortical white matter. Patients in whom the corpus callosum lesion disappeared had a better outcome than those in whom the lesion remained (good recovery/moderate disability; 92% vs 63%). The present MRI results suggest that some lesions in the corpus callosum following closed head injury are reversible, thus resembling edema that may be produced by a relatively mild shear strain force to the corpus callosum. PMID- 9416286 TI - Detection of lipid peroxidation and hydroxyl radicals in brain contusion of rats. AB - To examine the relationship between the free radicals and brain tissue damage, we investigated the intensity of brain hydroxyl (OH) radical generation and lipid peroxidation in the rat contusion injury model. A unilateral contusion was induced by a weight-drop method. All rats were decapitated six hours after the injury, and brain samples were taken from three portions (core, peripheral, and distal) to examine the specific gravity as an indicator of brain edema, generation of OH using an electron paramagnetic resonance spectrometer (EPR), and malondialdehyde (MDA) and 4-hydroxyalkenals production. Analysis of the specific gravity revealed cerebral edema on the ipsilateral side in the injured group. The signal intensity of EPR in the core and peripheral portions in the contusion group was significantly higher than that in the distal portion of the contusion group and that of all portions in the control animals. No significant difference was observed between the core and peripheral portions of the contusion group. The MDA and 4-hydroxyalkenals production was significantly higher in the core and peripheral portions than in the distal portion of the contusion group and that of all portions of the control group. The degree of posttraumatic brain edema was closely correlated with the increase of DMPO-OH adduct, MDA, and 4 hydroxyalkenals. These results support the current concept that free radical production following traumatic brain injury may induce lipid peroxidation and may be the direct cause of edema formation. PMID- 9416287 TI - Time course of tissue elasticity and fluidity in vasogenic brain edema. AB - We examined chronological changes in regional tissue elasticity (stiffness) and fluidity (1/viscosity) of the white matter during the development and resolution of vasogenic brain edema. Cryogenic injury was created in the cortex of cat brain, and the brain was prepared for measurement of regional tissue elasticity and fluidity. The results were then compared with the histology and tissue water content. Vasogenic edema developed in the white matter under the lesioned cortex (4-24 h) and was resolved by day 10. Regional tissue elasticity decreased significantly during the initial 24 h (45.3 +/- 32.5% of the control (mean +/- S.D.), and then increased to 158.6 +/- 32.3% of the control level at day 10. Regional tissue fluidity increased to 376.7 +/- 240.4% of the control level during the initial 24 h decreased to 77.7 +/- 17.9% of the control level at day 10. Histological examination of the white matter revealed widening of the inter fiber space at 4-24 h after lesioning and astrocytosis at day 10. Thus vasogenic edema causes an increase of tissue fluidity with a decrease of tissue elasticity. Reactive astrocytosis after the resolution of edema causes an increase of tissue elasticity with mild decrease in tissue fluidity. PMID- 9416288 TI - The penumbra zone of a traumatic cortical lesion: a microdialysis study of excitatory amino acid release. AB - A cortical tissue necrosis from focal trauma expands to 150% of its initial volume within 24 hrs. It is currently unknown, whether this phenomenon is part of the primary traumatic lesion or if it involves secondary mechanisms such as the release of excitatory amino acids into the traumatic penumbra zone. A microdialysis probe was inserted for that purpose in an oblique angle into the cortex of Sprague-Dawley rats, approximately 2 mm below the brain surface. One day later a highly standardized cortical freezing lesion was induced at the brain cortex above the microdialysis probe. Dialysate was collected prior to, during, and after trauma in 10 min intervals. In each animal, it was confirmed histologically, that the tip of the microdialysis probe was localized in the grey matter in close vicinity to the primary lesion. Following induction of the trauma a statistically significant increase of the dialysate level of aspartate, glutamate, glycine, and serine was observed, whereas that of alanine was not altered throughout the experiment. The posttraumatic increase of the excitatory neurotransmitters aspartate and glutamate indicates that these amino acids are involved in the secondary lesion growth after trauma. Confirmation of this hypothesis would require that specific antagonization of these excitotoxic amino acids is inhibiting growth of the lesion. PMID- 9416289 TI - Assessment of regional cortical blood flow following traumatic lesion of the brain. AB - A brain tissue necrosis from trauma gradually expands during the subsequent 24 h. Among others, deterioration of perifocal blood flow could be involved in the secondary extinction of initially viable brain tissue. A highly standardized freezing lesion was made in cerebral cortex of rats for frequent measurements of regional cortical blood flow with high spatial resolution by laser Doppler scanning flowmetry. Following trauma a profound decrease in cerebral blood flow was found, not only in the lesion proper but also in the perifocal and distant brain areas, which would support a role of ischemia in the secondary lesion growth. Further studies, however, are required, particularly on whether therapeutic improvement of perifocal flow is affecting expansion of the traumatic tissue necrosis. PMID- 9416290 TI - Comparison of the interleukin-6 and interleukin-10 response in children after severe traumatic brain injury or septic shock. AB - Inflammation may play an important role in the evolution of damage after traumatic brain injury (TBI). IL-6 and IL-10 are markers of inflammation that are pro- and anti-inflammatory in nature, respectively. They have been used as an index of the degree of inflammation in diseases including sepsis and meningitis. We hypothesized that both IL-6 and IL-10 would be increased in the cerebrospinal fluid (CSF) of children after TBI. We measured ventricular CSF concentrations of these metabolites (ELISA) each of the first 3 days after TBI in 15 children. CSF IL-6 was increased on day 1 (p < 0.05 vs days 2 or 3). CSF IL-10 was similarly increased on day 1 (p < 0.05). CSF IL-6 after TBI is similar to serum IL-6 levels previously reported in children with septic shock. In contrast, the CSF IL-10 response was markedly attenuated following TBI compared to sepsis. These data suggest a unique balance between pro- and anti-inflammatory cytokines in brain after TBI. PMID- 9416291 TI - Changes of MPO activity and brain water accumulation in traumatic brain injury experiments. AB - Comparison of brain tissue water content (BWC) data with myeloperoxidase activity assay (MPO) allows for analysis of the complex pathophysiological mechanisms of cerebral edema following catastrophic brain injuries. The neuroprotective effect of an experimental anti inflammatory drug (FL1003, butyrolactone) was tested in a traumatic brain injury (TBI) model using BWC and MPO analysis. We conducted these studies on a mini-pig model of severe TBI that is well characterized in our laboratory. The animals were divided into three animal groups: no injury, no treatment (control), injured and treated with FL1003, and injured, untreated with FL1003. They were maintained with fluids for 24 hours under general anesthesia. We employed the MPO assay to identify the degree of inflammatory cellular response (polymorphonuclear leukocytes, PMNLs) 24 hours following TBI and calculated brain density from the data of the gravimetric (Percoll) column method for BWC on brain samples. Our results demonstrated increased infiltration of PMNLs and a shift of water into the extravascular space in the injured animals. These changes were significantly (P < 0.05) attenuated in the animal group treated with FL1003. PMID- 9416292 TI - Hemodynamic depression and microthrombosis in the peripheral areas of cortical contusion in the rat: role of platelet activating factor. AB - Cerebrovascular damages leading to subsequent reductions in regional cerebral blood flow (rCBF) may play an important role in secondary cell damages following traumatic brain injury (TBI). Recent studies have demonstrated that rCBF markedly decrease in experimental model of TBI (e.g. fluid percussion injury, acute subdural hematoma, contusion). However, precise mechanisms underlying post traumatic CBF reduction remain unclear. In the present study, the rCBF changes and microthrombosis formation were investigated in a cortical contusional model in rats, and the effects of etizolam (platelet activating factor antagonist) on microthrombosis were tested. The rCBF in the peripheral areas increased transiently, and decreased to ischemic level 3 hours post- injury. The histological examinations revealed microthrombosis formation in the contused area, extending from the center to the peripheral areas within 6 hours post injury. The rCBF decrease and the contusion necrosis volume were significantly attenuated by etizolam administration. These results indicate that platelet activating factor is involved in microthrombosis formation and hemodynamic depression, and resultant ischemic damages within areas surrounding the contusion. PMID- 9416293 TI - Characterisation of brain edema following "controlled cortical impact injury" in rats. AB - Significance, origin and nature of posttraumatic brain edema are still being debated. Recently, a "controlled cortical impact injury" (CCII) was introduced to model traumatic brain injury. Purpose of this study was to investigate the development and nature of brain edema following CCII. Traumatic brain injury was applied to the intact dura of the left hemisphere in Sprague-Dawley rats (n = 52, 250-350 g b.w.). Ketamine/xylazine-anesthesia or inhalation-anesthesia were used. A pneumatic impactor with a diameter of 5 mm contused the temporo-parietal cortex with a velocity of 7 m/s and an impact depth of 2 mm. 24 hours post injury the brains were removed. Posttraumatic hemispheric swelling and water content were determined gravimetrically, Evans blue extravasation spectrophotometrically, area and volume of ischemia by staining with TTC. MRI studies were performed with T1 ,T2- and diffusion-weighted sequences. Posttraumatic swelling following CCII was 14.3 +/- 3.1%. Brain water content increased to 82.5 +/- 0.5% in lesioned hemisphere compared to 79.9 +/- 0.2% in control hemisphere. Following TTC staining, the average ischemic tissue volume was 56.7 +/- 19.2 mm3. There was a moderate uptake of Evans blue into the lesioned hemisphere. MRI studies demonstrated edema in 35.4 +/- 9.5 mm3 of the lesioned hemisphere. Gd-DTPA was taken up early after trauma only. A significantly decreased ADC (apparent diffusion coefficient) indicates the cytotoxic (ischemic) component of edema in this model. In conclusion, CCII produces significant posttraumatic brain swelling and edema which is both, of vasogenic and cytotoxic nature. Thus, the CCII models the human cortical contusion more appropriately and opens new avenues for therapeutical studies focussing on cortical contusions. PMID- 9416294 TI - The role of adenosine during the period of delayed cerebral swelling after severe traumatic brain injury in humans. AB - Cerebrovascular failure with an increase in cerebral blood volume or hyperemia contributes delayed cerebral swelling after severe traumatic brain injury (TBI) in humans. One mediator that could be involved in this process is adenosine, which stimulates a concurrent reduction in cerebral metabolic rate and an increase in cerebral blood flow (CBF). We hypothesized that during the delayed phase after TBI in humans: 1) CSF adenosine concentration is associated with uncoupling of CBF and CMRO2, and 2) adenosine formation is driven by mediator stimulated cAMP production in injured brain. We serially measured CBF and AVDO2, and CSF adenosine, lactate and cAMP after severe TBI in 13 humans. After 6-18 h, global CBF was increased and AVDO2 was reduced vs all other time periods, defining the uncoupling phase as the period between 18 h and 5 days. CSF adenosine concentration was negatively associated with AVDO2 and strongly associated with death (both p < 0.05), CSF lactate peaked during the initial 18 h, but remained increased for 5 days. CSF cAMP concentration was not increased (vs normal). The association between CSF adenosine concentration and death, and the correlation between uncoupling of CBF and oxidative metabolism and CSF adenosine concentration support our first hypothesis. In contrast, the low levels of cAMP in CSF observed in these patients, but persistently increased CSF lactate, refute our second hypothesis. We speculate that hyperglycolysis or occult ischemic foci are possible sources of ATP breakdown and adenosine formation, and that adenosine is playing a neuroprotective role. PMID- 9416295 TI - Near infrared spectroscopy (NIRS) in patients with severe brain injury and elevated intracranial pressure. A pilot study. AB - Near infrared spectroscopy (NIRS) was used to asses changes in regional cerebral oxygen saturation (rSO2) in 8 head injured patients with an intracranial pressure (ICP) higher or lower than 25 mmHg (n = 4 for each group). NIRS values in the high ICP group (> 25 mmHg) were significantly lower than in the low ICP group (< 25 mmHg). In contrast, arterial pO2, pCO2, peripheral oxygen saturation and transcranial Doppler sonography (TCD) values were similar in both groups. To further investigate changes in rSO2 to changes in peripheral oxygen saturation and arterial pO2, patients of both groups underwent an artificial hyperoxygenation (50% O2) period of 3 minutes. Both groups revealed similar values in peripheral oxygen saturation, arterial pO2, and TCD velocities at the end of the hyperoxygenation period. However, rSO2 values in patients with an ICP > 25 mmHg were significant lower than in patients with an ICP < 25 mmHg after the hyperoxygenation period. In addition, patients with an ICP < 25 mmHg revealed a significant increase in rSO2 values at the end of the hyperoxygenation period, not detectable in patients with an ICP > 25 mmHg. Our results suggest that NIRS may be an additional diagnostic tool in the non-invasive evaluation of impaired cerebral microcirculation in patients with increased intracranial pressure. PMID- 9416296 TI - MRI diffusion-weighted spectroscopy of reversible and irreversible ischemic injury following closed head injury. AB - The objective of this study was to detect the threshold between reversible and irreversible secondary insult of hypoxia and hypotension following closed head injury as measured by MRI. Adult Sprague rats were separated into 3 groups: I: Sham (n = 6), II: Trauma and hypoxia coupled with mild hypotension of 40-50 mmHg (n = 6), III: Trauma and hypoxia coupled with severe hypotension of 30-40 mmHg (n = 6). The measurement of brain water content (BWC) was based on T1, whereas the differentiation between reversible and irreversible secondary insult on the measurement apparent diffusion coefficient (ADC). The ADCs in both trauma and secondary insult groups decreased rapidly from a control level of 0.68 +/- 0.5 x 10(-3) to significantly different minimum levels of 0.52 +/- 0.5 x 10(-3) in Group II and 0.42 +/- 0.5 x 10(-3) mm2/second in Group III at 30 minutes. In Group II rats there was a complete recovery in ADC as well as in their clinical conditions, whereas ADC in Group III rats remained at the minimum level and the animals were brain dead. The BWC was also significantly different at four hours post injury (Group II: 80.3 +/- 0.7%, Group III: 81.8 +/- 0.8%). The data lead the authors to suggest that the threshold between reversible and irreversible posttraumatic secondary insult is very narrow, and the measurement of ADC can provide information that will enable the clinician to identify critical threshold beyond which recovery is not possible. PMID- 9416297 TI - Biphasic pathophysiological response of vasogenic and cellular edema in traumatic brain swelling. AB - The objective of this study was to quantify the temporal water content changes and document the type of edema (cellular versus vasogenic) that is occurring during both the acute and the late stages of edema development following closed head injury. Adult Sprague rats (n = 50) were separated into two groups: Group I: Sham (n = 8), Group II: Trauma (n = 42). The measurement of brain water content (BWC) was based on T1, whereas the differentiation of edema on the measurement of the random, translational motion of water protons (apparent diffusion coefficients-ADC) by MRI. In trauma animals, we found a significant increase in ADC (105%) as well as in BWC (0.7 +/- 0.3%) during the first 60 minutes post injury indicating vasogenic edema formation. This transient increase; however, was followed by a continuing decrease in ADC beginning at 45 minutes post injury and reaching a minimum at days 7-14 (-103%). Since the BWC continued to increase during the next day (10.3%), it is suggested cellular edema formation started to develop soon after injury and became dominant between 1-2 weeks post injury. In conclusion we may consider, that there is a predominantly vasogenic edema formation immediately after injury and later a more widespread and slower edema formation due to a predominantly cellular swelling. PMID- 9416298 TI - Intracranial pressure in a modified experimental model of closed head injury. AB - Intracranial pressure (ICP) was studied in a modified experimental model of closed head injury, in which the dynamic process of impact versus impulse loading was separately controlled. In this model, mortality of Wistar rats was considerably higher as compared to Sprague-Dawley rats subjected to similar traumatic conditions. Therefore Sprague-Dawley rats were used for all further experiments. Twenty-four rats, divided into 4 groups, underwent either sham or gradually increasing impact-acceleration trauma. Four hours after closed head injury, ICP measurements showed a significant correlation between the severity of the traumatic challenge and the resultant pressure rise (r2 = 0.731; p < 0.001). At the moment of impact there was a momentary blood pressure peak immediately followed by a transient period of hypotension. ICP measurements following directly to an impact-acceleration trauma, revealed an abrupt rise in ICP reaching pathological levels within 5 minutes. In conclusion, this modified model of closed head injury produces a predictable and reproducible pathologic ICP in Sprague-Dawley rats. PMID- 9416299 TI - Hypertonic/hyperoncotic saline reliably reduces ICP in severely head-injured patients with intracranial hypertension. AB - Hypertonic saline (HS) has been shown to decrease intracranial pressure (ICP) and cerebral water content in experimental models of traumatic brain injury (TBI). The purpose of the present study was to test the efficacy of administration of HS (7.5%) combined with 6% hydroxyethyl starch (molecular weight 200.000/0.60-0.66; HHES) for the treatment of therapy-resistant intracranial hypertension in patients with severe TBI. Six patients with severe TBI (GCS < 8) who met the inclusion criteria (therapy resistant ICP > 25 mmHg, cerebral perfusion pressure (CPP) < 60 mmHg, plasma-Na+ < 150 mOsm and > 4 hours since the last HS/HHES treatment) were prospectively enrolled in the study and received between one and ten bolus infusions of maximal 250 ml HS/HHES at a rate of 20 ml/min. A total of 32 infusions were given. Administration of HS/HHES significantly lowered ICP by 44% and improved CPP by 38% to well above 70 mmHg at 30 min without affecting arterial blood pressure or blood gases. Plasma sodium normalized within 30 min. Experimental studies from our laboratory indicate that the ICP lowering effect is primarily due to dehydration of brain tissue and that cerebral blood volume remains largely unaffected by HS. In summary, HS/HHES reduces otherwise therapy resistant intracranial hypertension and improves cerebral perfusion even after repeated administration without negatively affecting blood pressure or causing a rebound ICP increase. PMID- 9416300 TI - Topical application of insulin like growth factor-1 reduces edema and upregulation of neuronal nitric oxide synthase following trauma to the rat spinal cord. AB - The neuroprotective effects of insulin like growth factor-1 (IGF-1) on spinal cord injury induced edema formation, cell changes and profound upregulation of constitutive isoform of neuronal nitric oxide synthase (cNOS) was examined in a rat model. A focal spinal cord injury produced by making a lesion (about 2 mm deep and 5 mm long) of the right dorsal horn of the T10-11 segment resulted in a marked edema formation, cell injury and upregulation of cNOS following 5 h after trauma. In separate groups application of IGF-1 (0.1 microgram/microliter) topically on the exposed spinal cord (T10-11) starting from 30 min before injury (20 microliter), immediately before injury followed by 30 min, 60 min and thereafter every 1 h after injury until sacrifice resulted in significant attenuation of edema formation and cell changes. Immunohistochemistry showed a less pronounced expression of cNOS in the T9 and T12 segments of the cord in IGF treated rats compared to untreated traumatised controls. These results for the first time show that IGF treatment is neuroprotective and this effects of the IGF appears to be mediated via inhibition of NOS upregulation. PMID- 9416301 TI - Prostaglandins modulate constitutive isoform of heat shock protein (72 kD) response following trauma to the rat spinal cord. AB - The influence of prostaglandins on the constitutive isoform of heat shock protein (HSP 72 kD) response following a focal trauma to the rat spinal cord was examined using immunohistochemistry. A focal trauma to the spinal cord by making a longitudinal incision into the right dorsal (about 2 mm deep and 5 mm long) of the T10-11 segments, markedly enhanced the upregulation of HSP immunoreactivity in the traumatised as well as in the perifocal T9 and T12 segments. This upregulation of HSP was significantly reduced by pretreatment with indomethacin (10 mg/kg, i.p., 30 min before injury) an inhibitor of prostaglandin synthesis. These results show that blockade of PG synthesis prior to trauma has an inhibitory influence on the upregulation of HSP response, not reported earlier. PMID- 9416302 TI - The effect of endothelins on ion transport systems in cultured rat brain capillary endothelial cells. AB - Brain capillary endothelial cells regulate the movement of ions and water across the blood-brain barrier via specific ion transport systems. Disturbances in these ion transport systems are involved in the formation of ischemic brain edema. This study describes the effects of endothelins (i.e., ET-1 and ET-3) on ion transport systems in cultured rat brain capillary endothelial cells using 86Rb+ and 22Na+ as markers for K+ and Na+, respectively. ET-1 stimulated K+ uptake and efflux with EC50 values of 0.6 nM and 0.5 nM, respectively. The potencies of ET-3 on these responses were considerably lower. Both ET-1 and ET-3 stimulated Na+ uptake through a Na+/H+ exchange system with similar potencies (i.e., EC50 = 0.80 nM and 1.89 nM, respectively). ET-stimulated K+ uptake, K+ efflux, and Na+ uptake activities were all inhibited by BQ123 (selective ETA receptor antagonist). ET-1 stimulated K+ uptake and efflux, in contrast to Na+ uptake, were also reduced by protein kinase C inhibitors and by an intracellular Ca2+ chelator. The results suggest that ETs can affect the activities of ion and water transport at the blood-brain barrier through different signal transduction mechanisms. PMID- 9416303 TI - Neurotoxicity of serum components, comparison between CA1 and striatum. AB - In vasogenic brain edema, the neurotoxicity of extravasated serum components may contribute to neuronal damage. In the hippocampal CA1 sector and striatum, the neurotoxicity of serum was investigated. Rat serum was prepared as follows: Serum 1: whole serum, Serum-2: ultrafiltrated through a membrane with cut-off at molecular weight (MW) 100,000. Serum-3: through a membrane with cut-off at MW 20,000, and Serum-4: through a membrane with cut-off at MW 5,000. The infusion edema model was utilized for infusion of autologous serum into the brain. The brain tissue was histologically evaluated. The level of glutamate, total protein, and albumin was also measured in the sera used for infusion. The following results were obtained: 1) CA1 neurons were more vulnerable to all infused sera than striatal neurons, 2) there was a strong cellular response in the striatal site of infusion, but only a minimal in the CA1-sector, 3) the severity of damage of CA1 correlated with the glutamate concentrations of the infused sera, 4) further, there was a relationship between the degree of striatal neuronal loss and the amount of protein and albumin present in the infusate. It is, therefore, concluded that the neurotoxic properties of vasogenic edema fluid are also affected by specific features of the brain region of its extravasation. In addition, the pathological mechanisms associated with irreversible damage of neurons might be different in the CA1-sector and the striatum. PMID- 9416304 TI - Role of calcium ions in acidosis-induced glial swelling. AB - Tissue acidosis occurring in cerebral ischemia and traumatic brain injury is a mediator of cytotoxic brain edema. In vitro, extracellular lactacidosis induces swelling of glial cells in a dose dependent manner. pH-regulatory membrane transporters and channels have been identified which are involved in the increase of the glial cell volume. Underlying mechanisms of their activation are poorly understood, however. We have, therefore, addressed the question, whether and how Ca(2+)-ions play a role in acidosis-induced glial swelling and intracellular acidification. For that purpose C6 glioma cells were suspended and the pH in the medium was lowered from 7.4 (baseline) to 6.2 by isotonic lactic acid. Cell volume and intracellular pH (pHi) were assessed by flow cytometry. In the presence of Ca(2+)-ions the cell volume reached a maximum of 125.1% from acidosis. In experiments using a calcium-free suspension medium, cell swelling from acidosis was inhibited by 74%. Additional buffering of intracellular calcium (Ca2+i) had no further inhibitory effect on acidosis-induced cell swelling, while buffering of Ca2+i by BAPTA-AM alone did not affect the glial volume increase secondary to administration of lactic acid. pHi which was decreasing from acidosis was not affected by the experimental modifications of the Ca(2+) concentration in the medium or cytosol. The present data indicate that lactacidosis-induced glial swelling depends on the presence of extracellular Ca(2+)-ions, while release of Ca(2+)-ions from intracellular stores does not seem to be involved. PMID- 9416305 TI - Glutamate induced astroglial swelling--methods and mechanisms. AB - Glutamate (Glu) plays an important role in the early development of brain injuries caused by ischemia, i.e. stroke, or brain trauma. Glu induces a rapid astroglial swelling which, in turn, deranges the composition of neuroactive substances in the extracellular space. We report that Glu can induce astroglial cell swelling by interaction with metabotropic Glu receptors (mGluRs). Furthermore, the Na(+)-K(+)-2Cl- cotransporter, a Na(+)-K(+) ATPase, and the Na(+)-dependent electrogenic Glu carrier seem to be involved in this Glu-induced astroglial cell swelling. Two methods for studying cell swelling arc described. One is based on variations in the signal emitted by the fluorescent probe fura 2/AM when excited at its isosbestic point. These variations were shown to be directly proportional to variations in intracellular volume. Relative changes in cell volume and intracellular calcium concentration could be detected simultaneously in single astroglial cells. The other method used permits the cell volume to be calculated in relative terms with the aid of image processing techniques. PMID- 9416306 TI - Impaired learning of active avoidance in water-intoxicated rats. AB - Brain edema is an important clinical condition. Pathophysiological findings on behavioral changes may be helpful for a comprehensive understanding of brain edema. However, only few reports on behavioral studies of brain edema have so far appeared. Experiments using psychological techniques on animals are rather time consuming and may not be suitable for the study of transient conditions, as brain edema caused by trauma, vascular accidents, or others. We have developed a method for avoidance learning of rats using a running wheel apparatus with computer assistance. This model was employed in studies on brain edema from water introxication in rats. As a result, avoidance learning was significantly impaired by water intoxication. Either direct overhydration of the brain or indirect effects, as a decrease in cerebral blood flow, or both, are suggested as mechanisms underlying the impairment of behavior. PMID- 9416307 TI - Topical application of 5-HT antibodies reduces edema and cell changes following trauma of the rat spinal cord. AB - Involvement of serotonin in the early microvascular reactions and cell changes following trauma of the spinal cord was examined using topical application of serotonin antibodies to the traumatised cord in a rat model. A focal trauma of the spinal cord was produced by making an incision into the right dorsal horn of the T10-11 segments (about 2 mm deep and 5 mm long); the animals were allowed to survive 5 h after injury. Monoclonal 5-HT antibodies (1:20) were applied (25 microliters in 10 sec) to the traumatised spinal cord 2 min after injury. There was a significant reduction in the breakdown of the blood-spinal cord barrier, edema formation, and cell changes in the traumatised rats which received 5-HT antiserum compared to the injured rats given saline. These results show that 5-HT is an important mediator involved in the early pathophysiological responses of spinal cord injury. PMID- 9416308 TI - Steroids decrease uptake of carboplatin in rat gliomas--uptake improved by intracarotid infusion of bradykinin analog, RMP-7. AB - This study sought to determine whether dexamethasone (DXN) treatment of rats with intracranial gliomas would 1) further impair delivery of carboplatin to brain tumors, and 2) whether intracarotid infusion of the bradykinin analog, RMP-7, would improve delivery during concurrent DXN treatment. In DXN pretreated animals, 3 mg/kg/day of DXN was administered intraperitoneally for 3 days prior to Ki determinations. Ki of [14C] carboplatin into DXN-treated tumors and brain surrounding tumor (BST) was significantly lower compared to non-DXN treated tumors and BST (3.30 +/- 0.91 vs. 4.47 +/- 1.80, p < 0.05, and 0.94 +/- 0.84 vs. 2.18 +/- 0.79, p < 0.05, respectively). Intracarotid infusion of RMP-7 significantly increased the Ki for carboplatin in DXN-treated tumors (6.35 +/- 3.10 vs. 3.30 +/- 0.91, p < 0.01), however, RMP-7 increased Ki to a greater extent in tumors not pretreated with DXN (12.07 +/- 3.60 vs. 4.47 +/- 1.80, p < 0.0001). Dexamethasone decreases transport of carboplatin into brain tumors. Intracarotid infusion of RMP-7 selectively increases carboplatin transport to tumors. PMID- 9416309 TI - Effect of glycerol on blood flow distribution in tumoral and peritumoral brain tissue. AB - To evaluate the effect of glycerol, thirty-two patients with brain tumor were directed to the study, including 17 gliomas and 15 meningiomas. Blood flow before and after the administration of glycerol were measured with Xe CT. Glioma was significantly hypo-perfused. The peritumoral edema of glioma and meningioma were also hypo-perfused. On the other hand, Meningioma was significantly hyper perfused. After the administration of glycerol, blood flows were increased except for glioma. We suggested that, vascular responses to glycerol was different in the two tumor types. The steal phenomena of blood flow might occur in glioma. PMID- 9416310 TI - Assessment of vasoreactivity in brain edema by acetazolamide activation SPECT and PET. AB - Our study was performed to find out cerebrovascular reactivity post acetazolamide administration in patients with peritumoral edema. Adult patients (n = 9) underwent CBF measurement by 99mTc-HMPAO SPECT pre and post 1 gram i.v. acetazolamide. In all patients, this procedure was repeated once again within 10 days of performing tumor removal. Five of these patients also underwent CBF measurement pre and post 1 gram i.v. acetazolamide post surgery only using oxygen 15 labeled H2O PET. Asymmetry index (AI) was calculated as ratio of ROI counts in the peritumoral edematous area and symmetrical ROI on the contralateral normal hemisphere. The AI increased after acetazolamide in edematous gray matter post operatively though the resting AI remained almost same post operatively. AI in edematous white matter showed non-significant increase post operatively both at rest and after acetazolamide. Good linear correlation of AI between PET and SPECT was observed both in gray and white matter. The improvement of vascular reactivity in edematous gray matter after tumor removal suggests that mass effect not only reduces CBF but also suppresses vascular reactivity. White matter vascular reactivity in early post operative period is little improved, possibly due to factors other than mass effect i.e. excess water accumulation in white matter perivascular space. PMID- 9416311 TI - Tumor specific contrast enhancement study of Mn-metalloporphyrin (ATN-10)- comparison of rat brain tumor model, cytotoxic and vasogenic edema models. AB - ATN-10, Mn-metalloporphyrin, has been developed as a tumor selective contrast agent for magnetic resonance (MR) imaging. To investigate the tumor specificity of ATN-10, we produced three experimental in vivo models; rat bran tumor (9L glioma) model, vasogenic (cold injury) and cytotoxic brain edema (24-hour MCA occlusion) models. The time course of contrast enhancement was compared after intravenous injection of ATN-10 or Gd-DTPA, measuring the signal intensity of the region of interest. After ATN-10 administration, the 9L glioma model showed early (5 min) and delayed (24 hr-) peak enhancement whereas the cold injury model showed only early enhancement and the 24-hour MCA occlusion model did not show significant enhancement. After Gd-DTPA administration, all three models showed similar pattern of only early enhancement. As a contrast agent for MR imaging, ATN-10 showed different behavior than Gd-DTPA in demonstrating the blood-brain barrier disruption and moreover ATN-10 showed selective enhancement in experimental brain tumors. PMID- 9416312 TI - Apparent diffusion coefficient (ADC) and magnetization transfer contrast (MTC) mapping of experimental brain tumor. AB - Brain tumor tissue contains different pathological areas, such as tumor cell rich parts, necrotic tissues, and cyst. Furthermore, both neovascularization and edema formation progress along with the tumor progression. In this study we employed diffusion weighted (DW) and magnetization transfer contrast (MTC) imaging to chronologically investigate the biological characteristics of a rat glioma. RG-2 glioma cells were implanted stereotactically into the right hemisphere of male Wistar rats. MR images were taken 1, 2 and 3 weeks after inoculation. Apparent diffusion coefficient (ADC) and MTC values were calculated as follows; ADC = -ln (SI-DW/SI-T2)/1096, MTC = 1-SI-MTon/SI-MToff. Each mapping image was made based on the calculated average values of four pixels. The spatial signal changes and the real values were compared to the histological findings. The apparent increase of ADC was noted in the parenchyma adjacent to tumor suggesting the progression of edema. The tumor itself had similar or slightly increased ADC. Cystic and necrotic components appeared 2 weeks after implantation and they showed significantly higher ADC than those calculated in the contralateral putamen. On the other hand, MTC was slightly decreased in the parenchyma adjacent to the tumor, markedly within the tumor, and maximally in the cystic and necrotic area suggesting accumulation of macromolecules such as growth factors, cytokines, and serum albumin. PMID- 9416313 TI - The origin of lactate in peritumoral edema as measured by proton-magnetic resonance spectroscopic imaging. AB - Using in vivo proton-magnetic resonance spectroscopy (1H-MRS), which allows the measurement of metabolites of adequate tissue concentration, the origin of lactate in peritumoral edema has been assessed by comparison with lactate levels in the central and marginal areas of the tumor in 18 patients with cerebral gliomas. In the majority of cases lactate content in the area of peritumoral edema was lower than that in the tumor margin or tumor center, which is consistent with the assumption that the tumor is the source of lactate, which then reaches the surrounding area of edema by diffusion. In 3 of the 18 cases the amount of lactate in the peritumoral edematous tissue was higher than in the tumor, indicating that the lactate is locally produced on account of ischemia due to regional elevation of tissue pressure in the edematous area. PMID- 9416314 TI - Neuroprotective properties of a novel antioxidant (U-101033E) with improved blood brain barrier permeability in focal cerebral ischemia. AB - Many efforts have been undertaken to develop antioxidants against free radical induced brain damage, 21-aminosteroids, although accumulating in the cell membrane, thus protecting vascular endothelium from peroxidative damage hardly penetrate the blood-brain barrier. A novel group of antioxidants, the pyrrolopyrimidines, has a markedly improved ability to enter the brain parenchyma. In our current study the neuroprotective potential of the 21 aminosteroid U-74389G was compared with that of the pyrrolopyrimidine U-101033E in a rat model of reversible focal cerebral ischemia. Sprague-Dawley rats were subjected to unilateral occlusion of the middle cerebral artery with assignment to one of three treatment arms (n = 10 each), receiving either vehicle, U-74389G, or U-101033E. Regional CBF was recorded bilaterally by laser Doppler flowmetry. In addition, neurological examination was performed daily, with assessment of infarct volume at day seven. U-101033E reduced the infarct volume significantly by 51%, whereas U-74389G afforded non-significant attenuation only. U-101033E was found to improve neurological recovery promptly; animals with U-74389G began to recover only at the end of the experimental observation period. Differences in the regional CBF were not found in the contralateral hemispheres for either treatment group. We conclude that antioxidative compounds which cross the blood brain barrier are more effective in focal cerebral ischemia than agents which predominantly act on the endothelium of cerebral microvessels. PMID- 9416315 TI - Effect of alpha-trinositol on swelling and damage of glial cells by lactacidosis and glutamate. AB - The therapeutic efficacy of alpha-trinositol (D-myo-inositol-1,2,6 trisphosphate), an isomer of the intracellular messenger IP3, was analyzed for cytotoxic swelling and damage of glial cells in vitro from lactacidosis or glutamate. Lactacidosis and the interstitial accumulation of glutamate are prominent sequelae in ischemic or traumatic brain tissue. C6 glioma cells harvested from culture and suspended in a physiological medium were either exposed to pH 5.0 by administration of lactic acid, or to 1 mM glutamate at normal pH. Cell swelling and viability were quantified by blood flow cytometry. Addition of alpha-trinositol (3 mM) under control conditions at pH 7.4 resulted in transient cell shrinking to 96.5 +/- 1.3% of control within 3 min (p < 0.05). Lactacidosis of pH 5.0 led to an increase in cell volume to 139.7 +/- 1.3% within 20 min, whereas alpha-trinositol reduced the swelling response by approximately 25% (p < 0.01). In addition, cell viability was severely affected at pH 5.0 amounting to only 53.8 +/- 3.1% after 60 min. alpha-Trinositol was found to markedly improve cell viability; at 60 min 70.2 +/- 1.6% of the cells were still viable (p < 0.01). Addition of glutamate (1 mM) led to a steady increase in cell size, reaching 110% of control after 120 min, irrespective of wether alpha trinositol was present or not. The attenuation of cell swelling may be attributed to an interference with pH-regulatory mechanisms, such as the Na+/H(+) antiporter, while protection of cell viability might be caused be effects of alpha-trinositol on Ca(2+)-overload. On the other hand, the increase in cell volume by glutamate associated with its intracellular uptake was not influenced by alpha-trinositol. PMID- 9416316 TI - Effects of lecithinized SOD on contusion injury in rats. AB - To analyze the effect of lecithinized superoxide dismutase (SOD) on superoxide accumulation after traumatic injury, the expression of Cu,Zn-SOD mRNA was examined after contusion in rat using Northern blotting. As determined by specific gravity, lecithinized SOD decreased brain edema. The expression of Cu,Zn SOD mRNA increased at the core, peripheral and contralateral hemisphere of injury. These increases were then suppressed by lecithinized SOD. Our results support the hypothesis that superoxide may play an important role in edema formation after contusion, and that lecithinized SOD appears to prevent brain edema through a protective effect against superoxide injury. PMID- 9416317 TI - Extracellular changes of taurine in the peri-infarct zone: effect of lubeluzole. AB - Lubeluzole is a neuroprotective compound that has been shown to stereoselectively rescue sensorimotor function and reduce infarct size in a photochemical stroke model in rats. Tissue swelling, which occurs in the peri-infarct zone, is accompanied by a compensatory taurine release. Therefore, using a microdialysis technique, we aimed at measuring changes of extracellular concentrations of taurine in the peri-infarct zone and the effects of lubeluzole and its R-isomer. Lubeluzole blocked the increase of taurine in tissue immediately surrounding a photochemically induced thrombotic neocortical infarct. By contrast, the R-isomer was completely inactive. We hypothesize that lubeluzole may reduce osmoregulatory stress in peri-infarct tissue. PMID- 9416318 TI - Effect of tromethamine (THAM) on infarct volume following permanent middle cerebral artery occlusion in rats. AB - This study investigates the influence on tromethamine (THAM) on ischemic volume induced by permanent middle cerebral artery occlusion (MCAO) in rats. 14 male Sprague Dawley rats underwent left sided permanent MCAO by electro coagulation. Animals were treated either by 3-M THAM given intravenously in a single dosage of 0.6 mmol/kg body weight (THAM group: n = 7) 10 min following MCAO and again 1, 2, 3, 4 and 5 hours later or by NaCl 0.9% (placebo group: n = 7) in the same mode. Mean arterial blood pressure (MABP) was monitored for 30 min post MCAO and arterial blood gases were taken 10 min after the first injection. The extent of ischemia volume was assessed by planimetry of coronal sections stained with triphenyl-tetrazolium chloride (TTC) and with hematoxilin/eosin (HE). Tests for significance were accomplished by ANOVA on ranks. A difference of p < 0.05 was considered significant. The THAM group showed an insignificant decrease in MABP 1 min after injection (THAM: 75 +/- 11 mmHg, placebo: 86 +/- 10 mmHg). Arterial pH was significantly different (THAM: 7.46 +/- 0.04; placebo: 7.32 +/- 0.03). In TTC staining, the ischemia volume--given in absolute values and percentage of the total left volume--was significantly reduced in the THAM group (THAM: 43.9 +/- 8.3 mm3/7.0 +/- 1.3%; placebo: 95.2 +/- 13.8 mm3/14.2 +/- 2.0%). In HE staining, the reduction of ischemia, volume did not reach statistical significance (THAM: 49.1 +/- 9.9 mm3/9.6 +/- 1.8%; placebo: 66.3 +/- 14.5 mm3/13.1 +/- 2.8%). Based on these results, a moderate neuroprotective effect of THAM in experimental cerebral infarction could be demonstrated. PMID- 9416319 TI - Antioxidant, OPC-14117, attenuates edema formation and behavioral deficits following cortical contusion in rats. AB - Oxygen free radicals may contribute to tissue injury processes in the central nervous system following ischemia or trauma. Recent studies have suggested that inhibition of free radicals improves the outcome in experimental models involving such conditions, and antioxidant therapy appears promising. In the present study, behavioral changes and edema formation in rat cortical contusion model were investigated, and the effects of a superoxide radical scavenger, OPC-14117, were tested. Wistar rats were anesthetized with halothane inhalation. Cortical contusion was induced in the parietal cortex employing a controlled cortical impact device. Immediately following injury induction, OPC-14117 was administered (300 mg/kg, p.o.). Edema formation was assessed in the center and peripheral areas of the contusion by the specific gravity method. Behavioral changes were evaluated by the Morris water maze test and the habituation of exploratory activity. The results revealed that the vehicle-administered control showed progressive edema formation and behavioral deficits following the injury. These changes were significantly attenuated by the OPC-14117 treatment (p < 0.05). Further, OPC-14117 reduced the size of contusional necrosis (p < 0.05). These findings suggest that superoxide free radicals are involved in contusion-induced edema formation, necrosis formation, and behavioral deficits, and that OPC-14117 has a therapeutic potential to prevent secondary cell damage following traumatic brain injury. PMID- 9416320 TI - OPC-21268, an orally effective, nonpeptide arginine vasopressin V1 receptor antagonist reduces vasogenic brain edema. AB - We examined the effect of orally administered OPC-21268, a nonpeptide Arginine Vasopressin V1 receptor antagonist, on cold induced vasogenic brain edema in rat. Cold brain injury was induced by applying a copper rode cooled with liquid nitrogen for one minute. To mimic clinical use, one hour after induction of the cold lesion, rats were treated with orally administered OPC-21268 at doses of 100 mg, 200 mg, and 300 mg/kg every 8 hr for 24 hours. Two percent Evans blue in saline, in a volume of 1 ml/kg was given intravenously prior to cold injury. Twenty four hours after induction the cold lesion, brain water, brain tissue electrolytes, and plasma osmolality and electrolytes were measured. Quantitative evaluation of BBB permeability was performed using the Evans blue fluorescence method. The injury resulted in significant increases in the brain water and brain tissue sodium, and Evans blue concentration in both the lesioned and contralateral hemispheres (p < 0.01). OPC-21268 at doses of 200 mg and 300 mg/kg significantly decreased brain water and Evans blue concentrations in both the lesioned and contralateral hemispheres (p < 0.01). Brain tissue sodium content was significantly reduced at a dose of 300 mg/kg in the lesioned side (p < 0.05). There were no significant changes in other parameters throughout the experiments. Our results indicate that OPC-21268 exerts a protective effect in areas where the maximal amount of BBB breakdown occurs. PMID- 9416321 TI - Treatment of acute intracranial hypertension with RU 51599, a selective kappa opioid agonist. AB - RU 51599, a selective kappa opioid agonist which is a potent aquaretic, was studied for its effect on acute intracranial hypertension. In ketamine anesthetized cats acute intracranial hypertension was induced by progressive inflation of an epidural balloon with physiological saline at a constant rate of 0.5 ml/hr for three hours. In the control group (n = 8), the balloon was maintained inflated for another an hour after which it was deflated. In the post deflation period monitoring was continued for one hour. In the treatment group (n = 8), cats were treated by an intravenous (i.v.) injection of RU 51599, at a dose of 1 mg/kg every hour at the beginning of balloon inflation, during balloon inflation, at the completion of inflation, and after deflation. Changes in intracranial pressure (ICP), mean arterial blood pressure (MAP), cerebral perfusion pressure (CPP), blood gases, serum electrolytes and osmolality, and brain water content water content were studied in both groups. In the control group, epidural brain compression resulted in significant increases in the mean ICP up to 80.7 +/- 9.8 mmHg at 3 hrs (during balloon inflation), 68.6 +/- 8.3 after complete inflation, and 62.1 +/- 11.4 mmHg after deflation (P < 0.01), and significant decreases in CPP to 55.5 +/- 14.0 mmHg at 3 hrs (during balloon inflation), 43.0 +/- 11.2 mmHg after inflation, and 36.3 +/- 9.9 mmHg after deflation (P < 0.01). In the treatment group there were significant decreases in the mean ICP to 35.2 +/- 6.8 mmHg at 3 hrs (during balloon inflation), 32.3 +/- 7.9 after inflation, and 16.1 +/- 3.6 mmHg after deflation (P < 0.01), and significant increases in CPP to 103.2 +/- 6.1 mmHg at 3 hrs (during balloon inflation), 109.0 +/- 8.8 mmHg after inflation, and 102.7 +/- 8.2 mmHg after deflation (P < 0.01). Brain water content was also significantly reduced (P < 0.05). There were no significant changes in the serum electrolytes and osmolality throughout the experiments. Our results suggest that, the mechanism by which RU 51599 reduces ICP is related to reduction in the brain water content. PMID- 9416322 TI - The effect of nitric oxide inhibition on ischemic brain edema. AB - The involvement of nitric oxide (NO) in the development of ischemic cytotoxic edema was investigated by inhibiting nitric oxide synthase (NOS) activity with N omega-nitro-L-arginine (NLA). Bilateral carotid artery occlusion (15 min) alone or with release (15 and 60 min) served as a model for edema induction. NLA, N omega-nitro-D-arginine methyl ester (D-NAME) or Ringer's solution were administered 4 hr prior to ischemia or sham operation. Treatment with a stable nitroxide radical, 4-hydroxy-2,2, 6,6-tetramethylpiperidine-L-oxyl (TPL), was used to assess free radical involvement in edema. Accumulation of tissue water was evaluated by measuring specific gravity (SG) of brain cortex and histological examination. There was a greater reduction of cortical SG in early reperfusion (15 min) and a lesser decrease in SG (60 min later) in NLA-than in D-NAME- or Ringer's-treated gerbils. The NLA effect was confirmed by histological examination of the brain tissue. TPL treatment (pre- and postischemic) ameliorated the formation of edema to the same degree as NLA. The findings indicate a biphasic NLA modulation of cytotoxic edema most likely mediated through absence or presence of NO-derived free radicals. PMID- 9416323 TI - Blood-brain barrier disturbances after rt-PA treatment of thromboembolic stroke in the rat. AB - We studied the effects of rt-PA (recombinant tissue type-plasminogen activator) treatment on the blood-brain barrier (BBB) after thromboembolic stroke in rat. New MRI methods of diffusion and perfusion imaging to observe the hemodynamic and biophysical effects of thrombolysis were combined with methods for assessment of BBB disturbances. In untreated animals clot embolism produced a rapid drop in MRI perfusion values and the ADC (apparent diffusion coefficient), with subsequent infarction. BBB disturbances, visualised as extravasation of serum proteins on cryostat sections, were manifest in nearly all animals in the borderzone of infarcts. In animals treated with rt-PA 15 min after clot embolism thrombolysis resulted in reperfusion of affected brain regions with subsequent improvement of ADC values. Final lesion size on ADC maps was reduced by 36% relative to untreated animals. However, BBB disturbances were not improved after treatment. To the contrary, rt-PA treated animals showed further regions with serum protein extravasation in the infarcted territories and in distant non-ischemic brain regions. MR imaging with the BBB tracer GdDTPA showed more pronounced and widespread contrast enhancement in the rt-PA treated than in the untreated group. Increased blood-brain barrier disturbances have to be taken into account even when thrombolytic therapy is started very early after the onset of stroke. PMID- 9416324 TI - The effect of Dotarizine (Ca2+ channel blocker) on cerebral vessel reactivity in animals subjected to hyperventilation and anoxia. AB - Dotarizine--a novel piperazine derivative--belongs to wide spectrum Ca+ channel antagonists. It was reported to have strong vasodilatory and antiserotoninergic activities. Unlike other Ca+ channel blockers Dotarizine was found to have lower oral toxicity. In the presented study the influence of the oral administration of the novel compound on the blood flow velocity changes in different cerebral arteries--in basilar artery (BA) and middle cerebral artery (MCA)--was investigated under hyperventilation and hypoxic conditions of rabbits. In the first experimental group 25 mg/kg of Dotarizine dissolved in 0.25% agar was administered orally three times at the 10 hours' intervals. The sham group of animals was fed with agar of the same concentration. The results revealed that oral administration of Dotarizine diminished the vasoconstrictive effect of hyperventilation and this was more pronounced in MCA than in BA. During anoxic conditions stronger vasodilatory effects were observed in both groups of vessels and the low value of pulsatility index (PI) reflected pronounced decrease of the peripheral resistance, in comparison to the control group. Thus, the oral administration of Dotarizine decreases the peripheral resistance of cerebral vessels and therefore seems to have influence on the minute arteries of cerebrovascular system of the brain. PMID- 9416325 TI - Substance P endopeptidase activity in the rat spinal cord following injury: influence of the new anti-oxidant compound H 290/51. AB - The influence of the new antioxidant compound H-290/51 was examined on the substance P endopeptidase (SPE) activity in a rat model of spinal cord injury. This compound (H-290/51) has neuro-protective effects on edema and cell changes in this model. Infliction of trauma to the cord by making an incision into the right dorsal horn of the T10-11 segment resulted in a marked upregulation of SPE in the segments rostral to the lesion. On the other hand, the injured and adjacent caudal segments exhibited a marked down-regulation of the enzyme activity. Pretreatment with H 290/51 increased the SPE activity in the T9 segment but downregulated the enzyme activity in the T10-11 and T12 segments. The drug induced enzyme activity change was not further influenced by the trauma of the cord. The results indicate that a focal trauma induces widespread alterations in spinal cord SPE activity which can be influenced by the anti-oxidant drug H 290/51, suggesting that SPE is somehow involved in cell injury. PMID- 9416326 TI - Benzodiazepine receptors influence spinal cord evoked potentials and edema following trauma to the rat spinal cord. AB - The possibility that diazepam will influence spinal cord evoked potentials (SCEP), edema formation and cell changes following spinal cord injury (SCI) was examined in a rat model. The SCI was produced in equithesin anaesthetised animals by making a longitudinal incision (about 2 mm deep and 5 mm long) in the right dorsal horn of the T10-11 segments. The SCEP were recorded from the epidural space of the T9 segment after stimulation of the right tibial and sural nerves. The SCEP consisted of a small positive peak followed by a broad and high negative peak. Infliction of trauma to the rats resulted in an immediate and pronounced decrease of the maximal negative peak (MNP) amplitude. The spinal cord edema and cell changes were markedly pronounced 5 h after injury. Pretreatment with diazepam attenuated the early SCEP changes induced by the trauma and reduced the later development of edema and cell injury. These results suggest that benzodiazepine receptors are involved in trauma induced alterations in SCEP changes, edema formation and cell injury, not reported earlier. PMID- 9416327 TI - Induction of matrix metalloproteinases following brain injury in rats. AB - Matrix metalloproteinases (MMPs) are important in various pathophysiological processes related with the tissue remodeling. We planned the experiments to determine whether MMPs participate in disruption and repair of the tissue following brain injury. We have studied induction of MMP-9, a 92 kilodalton (kDa) gelatinase, in traumatic brain tissue, which may be produced by brain residual cells. We speculate that MMP-9 plays a role in post-traumatic brain edema formation. PMID- 9416328 TI - Expression of annexin and annexin-mRNA in rat brain under influence of steroid drugs. AB - Brain tissue of rats pretreated with methylprednisolone or with the 21 aminosteroid U74389F, and that of untreated control rats, was assessed for the expression of Annexin-1 (Anx-1) and the transcription of its mRNA. For this purpose Anx-1 cDNA was amplified and simultaneously a T7-RNA-polymerase promotor was incorporated into the cDNA using Polymerase Chain Reaction (PCR). Then digoxigenin-11-UTP was incorporated into the transcribed cRNA with T7-RNA polymerase. With this probe in situ hybridization was carried out in sections of the brain. The probe was visualized by an immunoassay using an anti-digoxigenin antibody conjugate. Anx-1 protein was assessed by means of immunohistochemistry using a polyclonal antibody. The various brain areas of the control animals showed an appreciable amount of Anx-1 at mRNA or protein level; on the other hand, the animals which had been pretreated with either steroid, showed a more intense Anx-1 mRNA signal than the controls in many areas. In the pretreated animals Anx-1 immunostaining was unchanged in cortex, basal ganglia, amygdala and septum, but more intense in hippocampus, hypothalamus and thalamus. In ependyma, choroid plexus, meninges, and vascular walls there was no Anx-1 mRNA transcription detectable. An opposite profile was shown by the Anx-1 immunoreactivity, the protein was present in control animals as well as the steroid-pretreated animals, suggesting that here the protein was either from systemic origin, or has diffused from adjacent structures. The results indicate that Anx-1 mRNA transcription is upregulated by either steroid, and that in the untreated animals there is a resting level of Anx-1 mRNA transcription, presumably reflecting physiological influences on Anx-1 expression. PMID- 9416329 TI - Role of protein kinase C in acidosis induced glial swelling--current understanding. AB - A major factor in secondary brain injury following cerebral trauma is accumulation of lactic acid resulting in glial swelling. Further, evidence obtained in this context demonstrates activation of protein kinase C (PKC) under these circumstances. Glial swelling from acidosis is attributable to activation of the Na+/H(+)-exchanger, mediating influx of Na(+)-ions in exchange for the extrusion of H+ ions. The antiporter is activated following phosphorylation by PKC. The current study was made to elucidate the role of PKC activation in acidosis-induced glial swelling. For that purpose, suspended C6 glioma cells were used to examine changes of the cell volume and intracellular pH (pHi). Acidosis was induced by administration of isotonic lactic acid. Stimulation of PKC by the phorbol-ester PMA was significantly enhancing glial swelling from severe acidosis (pH 6.2), whereas the decrease of pHi was somewhat attenuated. On the other side, inhibition of PKC by staurosporine did not affect cell swelling nor the decrease of pHi from acidosis. The results indicate that activation of PKC in cerebral trauma or ischemia may enhance glial swelling from lactacidosis. PMID- 9416330 TI - Dependence of basal cerebral blood flow and cerebral vascular resistance in spontaneously hypertensive rats upon vasoconstrictor prostanoids. AB - The effects of indomethacin (inhibitor of cyclooxygenase), imidazole (inhibitor of thromboxane A2 synthase) and SQ 29548 (antagonist of TxA2/PGH2 receptors) on basal CBF and CVR were studied in normocapnic and normoxic SHR and WKY rats. CBF was measured by the intracarotid 133Xe technique. CVR was calculated as ratio of mean arterial blood pressure and CBF. Resting CBF did not differ between SHR and WKY. MABP and CVR were significantly higher (p < 0.01) in SHR than in WKY. Indomethacin (6 mg/kg, i.v.) produced a significant long-lasting decrease of CBF and increase of CVR in both strains, although these effects were more pronounced (p < 0.01) in WKY. Imidazole (20 mg/kg, i.v.) had no effect on measured variables in either strain. SQ 29548 (1 mg/kg, i.v.) produced a significant increase of CBF in SHR (p < 0.001) but not in WKY. CVR decreased in SHR parallel to the increase of CBF but remained unchanged in WKY. Our results demonstrate that, in contrast to WKY, basal CBF and CVR in SHR depend upon vasoconstricting prostanoids which act on TxA2/PGH2 receptors but are distinct from thromboxane A2. PMID- 9416331 TI - The mechanism of reversible osmotic opening of the blood-brain barrier: role of intracellular calcium ion in capillary endothelial cells. AB - Despite clinical and experimental interest in the osmotic opening of the blood brain barrier (BBB), the mechanism underlying the phenomenon remain undetermined. The aim of this study is to investigate the mechanism of intracellular Ca2+ change in brain microvascular endothelial cells subjected to hyperosmotic stress. Cultured rat brain capillary endothelial cells were obtained by two-step enzymatic purification. Intracellular Ca2+ was measured by a confocal laser scanning microscope. After exposing the endothelial cells to 1.4 M mannitol for 30 seconds, the change of intracellular Ca2+ concentration was monitored. Intracellular Ca2+ concentration increased rapidly and reached its peak value within 10 seconds after the application of mannitol. The Ca2+ concentration returned to the basal level within 200 seconds. A calcium channel blocker nifedipine (100 microM, 10 microM) did not block the increase. A specific blocker (KB-R7943) of Na+/Ca2+ exchange did not affect the rapid elevation of intracellular Ca2+. However, it blocked the return phase almost completely. The results indicated that the Na+/Ca2+ exchanger pumped out the increased intracellular Ca2+ during the return phase. Reversible osmotic disruption and reconstruction of the BBB is not due to simple mechanical shrinkage of the endothelial cells but is due to the intracellular Ca(2+)-activated complex mechanism. The manipulation of the reconstruction phase, which depends on Na+/Ca2+ exchanger, may have clinical implications. PMID- 9416332 TI - Blood-brain barrier disruption, edema formation, and apoptotic neuronal death following cold injury. AB - The temporal pattern of brain edema and apoptosis following cold injury was investigated. Extravasation of Evans blue from the disrupted blood-brain barrier (BBB) maximized immediately after injury and returned to the control level at 24 h. However, water content increased up to 24 h and was maintained at a higher level than the control at 72 h. Apoptotic cells as detected by in situ end labeling were observed in the entire lesion at 24 h. At 72 h after injury, these apoptotic cells were observed in the margin of the lesion, but not in the core. These results suggest that apoptosis contributes to neuronal damage following cold injury and may result from the development of vasogenic edema. PMID- 9416333 TI - Blood-brain barrier disruption, HSP70 expression and apoptosis due to 3 nitropropionic acid, a mitochondrial toxin. AB - 3-Nitropropionic acid (3-NP), a mitochondrial toxin, induces apoptosis in the striatum. We wanted to determine if there was a relationship between mitochondrial dysfunction, disruption of the blood-brain barrier (BBB), and apoptosis. BBB disruption following intrastriatal injection of 3-NP was assessed by Evans blue leakage, brain water content, and by the expression of the 70 kDa heat shock protein (HSP70) and mRNA. Apoptosis was assessed by in situ terminal deoxynucleotidyl transferase-mediated uridine 5'-triphosphate-biotin nick end labeling (TUNEL) and gel electrophoresis to detect internucleosomal DNA fragmentation. Microscopic evidence of Evans blue leakage due to 3-NP was present only 3 hr after injection. Both internucleosomal DNA fragmentation and TUNEL labeling did not appear until 24 hr after injection. HSP70 (protein and mRNA) was also elevated by 24 hr. There was a quantitative increase in Evans blue leakage and brain water content due to 3-NP by 3 days after injection. Our results suggest that BBB disruption is an early event followed by increased HSP70 expression and apoptosis. We speculate that 3-NP damages endothelial cells, leading to vasogenic edema and apoptosis. PMID- 9416334 TI - Blood-brain barrier breakdown occurs early after traumatic brain injury and is not related to white blood cell adherence. AB - The time course of blood-brain barrier (BBB) breakdown after traumatic brain injury (TBI) has important implications for therapy. This study was conducted in order to test post-traumatic BBB dysfunction in a model of fluid-percussion induced TBI in rabbits at 1 and 6 hours after TBI and relate it to white blood cell (WBC) activation. Ten anesthetized rabbits had chronic cranial windows implanted three weeks prior to experimentation. Fluid-percussion injury (3.5 atm.) was induced and animals were followed for 1 or 6 h. Intravital fluorescence videomicroscopy was used to assess BBB permeability and WBC adhesion to pial venules. Na(+)-fluorescein was infused continuously over 30 min at either 30 min (Group I, n = 5) or 5.5 h (Group II, n = 5) after TBI. Microvascular permeability in individual postcapillary venules was assessed qualitatively at 1 and 30 min after start of infusion. TBI led to a transient mean arterial blood pressure (MAP) surge after trauma and a progressive increase in the number of sticking WBCs per mm2 vessel wall. Na(+)-fluorescein extravasation was observed in 4 out of 5 Group I animals and in none of Group II. BBB breakdown was not associated with WBC sticking. We conclude that after fluid-percussion injury the BBB is damaged at 1 h post-trauma and that its function is restored 6 h later. Increased WBC sticking at 6 h is not associated with BBB breakdown. Whether WBCs may cause vascular permeability changes at a later point needs further investigation. PMID- 9416335 TI - Acute blood-brain barrier changes in experimental closed head injury as measured by MRI and Gd-DTPA. AB - The objective of this study was to determine the early time course of blood-brain barrier (BBB) changes in diffuse closed head injury (CHI) and to what extent BBB is affected by secondary insult. The BBB disruption was quantified using T1 weighted MRI following administration of Gd-DTPA. The maximal signal intensity (SI) enhancement was used to calculate BBB disruption. A new CHI model was used to induce injury. Adult SD rats were separated into four groups: Group I: Sham (n = 4), II: Hypoxia and Hypotension (HH, n = 4), III: Trauma alone (n = 23), and IV: Trauma coupled with HH (THH, n = 14). Following trauma, a 30 minute insult of hypoxia (PaO2 = 40 mmHg) and hypotension (MABP = 30 mmHg) were imposed. In trauma animals, SI increased dramatically immediately following impact. By 15 minutes, permeability decreased exponentially and by 30 minutes was equal to that of control. In THH animals, SI enhancement was lower after the trauma, consistent with reduced blood pressure and blood flow. However, the SI increased dramatically upon reperfusion and was equal to that of control after 60 minutes. In conclusion we may consider, that CHI is associated with a rapid and transient BBB opening which begins at the time of the trauma and lasts not more than 30 minutes. It has been also shown that addition of hypoxia and hypotension prolongs the time of BBB breakdown. PMID- 9416336 TI - Transport of human beta-amyloid peptide through the rat blood-brain barrier after global cerebral ischemia. AB - In an attempt to produce an animal model of the Alzheimer's disease (AD), beta amyloid-(1-42)-peptide (beta A1-42) was injected into the femoral vein in rats after single and repeated cardiac arrest (CA). After survival of 3.5 months, the brains immunoreactivity was evaluated using light microscopic immunocytochemistry of monoclonal beta-amyloid peptide (beta A) antibody 4G8 (mAb 4G8). Rats receiving beta A1-42 after CA demonstrated multifocal and widespread extravasation of beta A1-42 in extra- and intracellular space. The permeability to beta A1-42 was significantly higher in rats after repeated cerebral ischemia. As in AD, there were irregular diffuse amyloid plaque-like deposits and neuronal loss with reactive gliosis. Our data in ischemic rats with beta A1-42 represent a novel animal model of Alzheimer's pathology. PMID- 9416337 TI - Enhanced brain opioid receptor activity precedes blood-brain barrier disruption. AB - We studied the effects of transient postischemic increased opioid receptors (OPR) binding (mu, delta, kappa) on blood-brain barrier (BBB), brain water content and brain mitochondrial oxidative enzymes system. Cats were exposed to temporary middle cerebral artery occlusion (MCAO). The significant increased OPR bindings observed 10 min after the release of MCAO (ischemic rCBF = 7 +/- 1 to 11 +/- 2 ml/100 g/min) preceded the early and late BBB disruptions, brain edema and postischemic impaired mitochondrial oxidative enzymes functions. Further, the study suggests indirectly that the latter process was irreversible and hence associated with subsequent ischemic cerebral infarction. In addition, the results revealed a possible viable therapeutic window in the early postischemic recirculation period, before the onset of impaired mitochondrial oxidative function. PMID- 9416338 TI - The effects of methylmethacrylate's hyperthermic polymerization on cerebral vascular permeability. AB - This study was undertaken to analyze the effects of significant hyperthermia (> 100 degrees C) associated with the polymerization of polymethlymethacrylate (PMM) on the permeability of the cerebral vasculature in rats. The method used to visualize the pial vasculature included the open pial window technique and epifluorescence microscopy. Results indicated that there is a significant increase in cerebral vascular permeability following in situ polymerization of PMM over the craniectomy site. PMID- 9416339 TI - Intraischemic hypothermia during pretreatment with sublethal ischemia reduces the induction of ischemic tolerance in the gerbil hippocampus. AB - We examined whether mild brain hypothermia during pretreatment with sublethal 2 min ischemia affected the tolerance to subsequent lethal 5-min ischemia. The neuronal densities in the hippocampal CA1 sector of gerbils preconditioned at mild brain hypothermia (32% of normal) were significantly lower than those in gerbils preconditioned at brain normothermia (70% of normal). 72-kDa heat-shock protein immunoreactivity in the CA1 sector preconditioned at mild hypothermia was reduced. These results suggest that mild brain hypothermia during pretreatment with sublethal ischemia reduces the tolerance to subsequent lethal ischemia. PMID- 9416340 TI - The relationship between brain temperature and neutrophil accumulation after traumatic brain injury in rats. AB - Mild hypothermia reduces secondary damage after traumatic brain injury (TBI) in rodent models; however, the mechanisms involved in this beneficial effect remain unclear. We previously reported that TBI induces the upregulation of adhesion molecules and infiltration of neutrophils (PMN) in brain. Since PMN accumulation may be associated with the development of hyperemia and blood-brain barrier injury, we hypothesized that hypothermia would reduce acute inflammation after TBI in rats. To test this hypothesis, rats were anesthetized and subjected to TBI by controlled cortical impact to left parietal cortex. Brain temperature was controlled at 32 degrees C, 37 degrees C, or 39 degrees C (n = 8 per group) for 4 h after TBI, then rats were sacrificed and brain were harvested. Immunohistochemistries were performed on brain sections using antibodies that recognize the adhesion molecules E-selectin and intercellular adhesion molecule-1 (ICAM-1), and PMN. PMN were also quantified using a myeloperoxidase (MPO) assay. PMN accumulation in injured brain was decreased in rats maintained at 32 degrees C vs 39 degrees C (4-fold by immunohistochemistry and 8-fold by MPO, p < 0.05). E selectin was induced after TBI, but not attenuated by hypothermia. ICAM-1 was not up-regulated at this early time after TBI. Based on these preliminary data, we conclude that mild hypothermia reduces PMN accumulation in injured brain during the initial 4 h after TBI, without decreasing adhesion molecule expression. PMID- 9416341 TI - Effect of mild and moderate hypothermia on the acidosis-induced swelling of glial cells. AB - The effect of mild (32 degrees C) and moderate (27 degrees C) hypothermia was analyzed on the cell volume and intracellular pH (pHi) of C6 glioma cells at normal pH and during lactacidosis at pH 6.2 in vitro. The cells were suspended in an incubation chamber under continuous control of pH, PO2 and temperature. Cell swelling was quantified by an advanced Coulter-system. pHi was measured by flow cytometry using the fluorescent dye bis-carboxyethyl carboxyfluorescein (BCECF). Following a control period at 37 degrees C, the ambient temperature was decreased to 32 degrees C for 30 min, and subsequently to 27 degrees C for another 30 min. Hypothermia alone led to an immediate and significant cell volume increase of 107.3 +/- 0.4% (mean +/- SEM) of control after 30 min at 32 degrees C, and further swelling to 110.5 +/- 0.9% after 30 min at 27 degrees C. Yet, hypothermia (27 degrees C) afforded partial protection against the acidosis-induced cell swelling at pH 6.2, which was reaching to 120.4 +/- 0.9% in the normothermic control group after 60 min, while only to 111.3 +/- 0.9% at 27 degrees C. Hypothermia, however, was associated with a more pronounced decrease of the pHi during acidosis (6.3 +/- 0.04) as compared to that of the normothermic control falling then to 6.5 +/- 0.03. The results demonstrate that mild and moderate hypothermia induce glial cell swelling, but simultaneously inhibit cell swelling from acidosis. The protection against cell swelling, however, has its price as indicated by the enhancement of the intracellular acidification. PMID- 9416342 TI - Cerebral vascular response to hypertonic fluid resuscitation in thermal injury. AB - The purpose of this project was to study the effects of various resuscitation fluid protocols in a systemically thermally injured rat sustaining 70% body surface area third degree burn using the pial window model in rats. The results show that there was a significant albumin leak in the cerebral vessels in both the experimental group which underwent no resuscitation fluid, as well as the experimental group that was resuscitated with Lactated Ringer's solution using the Parkland formula. When this was compared to the control group, as well as to the experimental group which received hypertonic hyperosmotic saline (HMS) boluses every hour, there was little if any leakage seen. PMID- 9416343 TI - Morphologic analysis of the cerebral microcirculation after thermal injury and the response to fluid resuscitation. AB - Using the pial window model, we have previously demonstrated that there is a disruption of the blood brain barrier with distal thermal injury [1-3]. Our laboratory has shown that treatment with Lactated Ringer's Solution did not improve labeled albumin leakage. However, treatment with hypertonic hyperosmotic saline (HHS) solution post thermal injury seemed to essentially eliminate the albumin leakage in cerebral vessels. Using adult Sprague-Dawley rats and epifluorescent microscopy, the cerebral vessel size and diameter were measured, as well as the number of leukocytes rolling or adherent to the endothelium. The results show that there was significant progressive arterial dilatation over six hours in the thermally injured animals treated with HHS. There was also a significant increase in leukocyte number if the animals were thermally injured and had no resuscitation fluid or if the animals were thermally injured and underwent resuscitation fluid with Lactated Ringer's compared to either the control group or the group that was treated with HHS after thermal injury. PMID- 9416344 TI - Role of neurochemicals in brain edema and cell changes following hyperthermic brain injury in the rat. AB - The involvement of three potent neurochemical mediators of the edema formation such as serotonin, prostaglandins and opioids in the pathophysiology of hyperthermic brain injury was examined in a rat model using a pharmacological approach. Hyperthermic brain injury was induced in conscious young rats by exposing them to heat stress at 38 degrees C for 4 h. In these rats the blood brain barrier (BBB) permeability, brain edema, cerebral blood flow (CBF), heat shock protein 72 kD (HSP) response and cell changes were examined. Pretreatment with ketanserin (a serotonin-2 receptor antagonist), indomethacin (prostaglandin synthesis inhibitor) and naloxone (opioid receptor antagonist) in separate groups of rats reduced hyperthermia and HSP response following heat stress and significantly attenuated changes in the BBB permeability, brain edema, CBF and cell reaction. These results suggest that the pathophysiology of hyperthermic brain injury is a complex mechanisms and several neurochemicals are involved in the brain pathology caused by heat stress. PMID- 9416345 TI - Acute heat stress induces edema and nitric oxide synthase upregulation and down regulates mRNA levels of the NMDAR1, NMDAR2A and NMDAR2B subunits in the rat hippocampus. AB - The influence of heat stress on constitutive isoform of neuronal nitric oxide synthase (cNOS) and NMDA receptor gene expression in hippocampus was examined in a rat model. Subjection of animals to 4 h heat stress at 38 degrees C resulted in a marked upregulation of cNOS in the hippocampus accompanied with a marked general expansion and edematous cell changes. On the other hand NMDA receptor messenger RNA encoding NMDAR1, NMDAR2A and NMDAR2B subunits showed a marked downregulation in the hippocampus of heat stressed rats compared to the controls. Our results show that upregulation of cNOS is instrumental in heat stress associated edema and cell injury. Furthermore, an increased production of NO as evident with upregulation of cNOS appears to be a key factor in the downregulation of NMDA receptor gene expression in heat stress. PMID- 9416347 TI - Effect of infection brain edema on NMDA receptor binding in rat brain in vivo. AB - The purpose of this study was to determine the effect of infection brain edema (IBE) in the rat model induced by injecting pertussis bacilli (PB) into the left carotid artery. The specific binding of N-methyl-D-aspartate (NMDA) receptor with [3H] MK-801 was measured in the neuronal membrane of cerebral cortex. The Scatchard plots were performed. The Bmax values were 0.623 +/- 0.082 and 0.606 +/ 0.087 pmol/mg protein in the group that received normal saline (NS) and PB respectively (P < 0.05). The Kd values were 43.1 +/- 4.2 and 30.5 +/- 3.0 nM in the groups NS and PB respectively. The results indicated that the affinity of NMDA receptor was significantly higher in the group PB than group NS, whereas the total number of NMDA receptors had not changed in the IBE model. The increase of affinity of NMDA receptor can be blockaded by MK-801 pretreatment in vivo. PMID- 9416346 TI - Choroid plexus ion transporter expression and cerebrospinal fluid secretion. AB - The Cl-/HCO3- exchanger (AE2 isoform) and the Na+/K(+)-ATPase at the choroid plexus are both thought to be involved in CSF secretion. However, both transport mechanisms are also postulated to have a role in CSF ion homeostasis raising questions as to which parameters control the expression of these transporters? Northern blots have been used to assess AE2 mRNA levels in rats subjected to alterations in blood pH or blood osmolality (a factor affecting CSF secretion). Six hours of alkalosis induced a 40% increase in AE2 mRNA (p < 0.01), suggesting that alterations in the expression of this transporter play a role in CSF pH homeostasis. In contrast, changes in osmolality did not affect AE2 mRNA. Western blots of Na+/K(+)-ATPase subunits were also examined to determine whether hypo and hyperkalemia affect protein levels of this transporter. There was a positive correlation between the plasma K+ concentration and both alpha 1- and beta 1 subunit protein levels suggesting a role for this transporter in CSF K+ homeostasis. As changes in plasma K+ and pH affect choroid plexus ion transporters but do not appear to alter CSF production, these results suggest the presence of compensatory mechanisms. Understanding of such mechanisms may facilitate therapeutic control of CSF production. PMID- 9416348 TI - The rapid flow of cerebrospinal fluid from ventricles to cisterns via subarachnoid velae in the normal rat. AB - 14C-sucrose in 0.5 microliter of buffered saline was infused over 30 sec into one lateral ventricle, and its subsequent distribution was determined in brain, meninges, cerebral blood vessels, and cerebrospinal fluid (CSF) by quantitative autoradiography. Within 3.5 min, infused radiotracer had moved into the third ventricle, the velum interpositum (an extension of the subarchnoid system that contains many blood vessels), the aqueduct, the mesencephalic and fourth ventricles, and the superior medullary velum (a part of the subarachnoid system that touches the mesencephalic and fourth ventricles). The CSF within both of these velae appears to empty into the quadrigeminal and ambient cisterns. Within 5 min radioactive sucrose was also found in the interpeduncular cistern. About 15% of the injected sucrose quickly left the ventricles and entered these large cisterns. In contrast to most CSF-brain interfaces, little sucrose moved from CSF into the medulla next to the lateral recesses and tissues such as the superior colliculus that lie adjacent to the large CSF cisterns. A thick, multilayered glia limitans visible on electron micrographs seemed to form a CSF-brain barrier at these interfaces. Some of the infused 14C-sucrose persisted in the perivascular spaces and walls of arteries and arterioles for more than 3.5 hr. These findings suggest that CSF may function to deliver various agents and factors to pial and parenchymal arteries and arterioles. PMID- 9416349 TI - Quantitative analysis of brain edema resolution into the cerebral ventricles and subarachnoid space. AB - Resolution of vasogenic brain edema was examined using a model of infusion of fluid into the brain of rabbits. For this purpose infusion of Texas Red-albumin (MW 67.000 D) and sodium fluorescein (MW 376 D) dissolved in artificial cerebrospinal fluid (mock CSF) was made into the white matter of the left frontal lobe of the brain. In order to quantify the portion of edema fluid which was cleared by the ventricular system, a ventriculo-cisternal perfusion was performed with mock CSF. A closed cranial window was implanted above the left parietal brain for superfusion of the cerebral cortex with mock CSF, in order to study resolution of the artificial edema fluid via the subarachnoid space. CSF-samples were collected in 30 minutes-intervals and analysed with a spectrophotometer. The clearance of edema fluid was examined under low (2-5 mmHg) and medium (9-12 mmHg) intracranial pressure (ICP). In the low-pressure group both edema fluid markers were found in the ventriculo-cisternal and subarachnoid perfusate at 60 min and 90 min, respectively, after start of infusion. In the group with moderately increased ICP the markers appeared at 90 min and 120 min, respectively. The amount of clearance of fluorescent dye via the subarachnoid space was the same in both groups and independent of the intracranial pressure. PMID- 9416350 TI - Responses of cerebral blood flow regulation to activation of the primary somatosensory cortex during electrical stimulation of the forearm. AB - We assessed the cerebral blood flow (CBF) response to electrical stimulation of the contralateral forearm over the primary somatosensory cortex (S-I) in anesthetized cats. CBF was monitored continuously using laser-Doppler flowmetry (LDF). In the first set of experiments, the effects of varying stimulus frequency and intensity were examined. During stimulation, CBF in S-I was increased significantly. At high stimulus intensity, response reached a near-plateau level. In the second set of experiments, the CBF response after introduction of an intracerebral mass was investigated using a mechanical microballoon model to simulate an intracerebral hematoma. A microballoon was inserted into the ventral posterolateral nucleus of the thalamus (VPL). Following gradual balloon inflation, there was a rapid reduction in CBF response. We conclude that CBF regulation to neuronal activation is affected by stimulation parameters, and is impaired by an intracerebral mass obstructing the afferent sensory pathway. PMID- 9416351 TI - A new model of spinal cord edema. AB - Edema of the spinal cord has not been well understood. Brain edema produced by Marmarou's infusion method is essentially similar to vasogenic edema. This infusion method for producing edema was applied to a cat spinal cord. After laminectomy, a 30-gauge needle was inserted into the intumescentia cervicalis. A total amount of 10 microliters of 2% Evans' blue or autoserum were infused using an infusion pump at a rate of 5 microliters/hr. Macroscopally, Evans' blue was observed in the vicinity of infused site at the same level of the needle insertion and was seen spreading mainly longitudinally in the lateral column for a certain distance. The extracellular space was markedly distended in the in fused white mater and filled with electron-dense materials which were thought to be proteins in the electron microscopic study. The fine structural features were similar to the findings which were seen in Marmarou's infusion type of brain edema. Using this model, it seems to be feasible to produce reproducible spinal cord edema at any location in order to investigate not only the morphological aspect but also physiological aspect of the edema. PMID- 9416352 TI - Assessment of the CAMINO intracranial pressure device in clinical practice. AB - The purpose of this study was to investigate reliability, handling characteristics and complication rate of the CAMINO-ICP-monitor-system in clinical routine. In a case controlled study 82 patients with intracranial pathology necessitating ICP-monitoring received either a ventricular or a parenchymal CAMINO-device. Clinical assessment of curve shape and apparent reliability of the measurement was documented. Probe position and presence of hematoma was evaluated in all patients with a CT after probe insertion. Handling complications, i.e. dislocation were recorded. At the end of the measuring period the drift of the probe was checked ex vivo and a two point calibration was performed using a water column. During one year 82 patients received 95 probes (parench, 73. ventric. 22). The average measuring period was 91.3 +/- 70.6 hrs. Catheter position was verified by CCT for 67 (70.5%) probes. 92.5% of the devices were placed correctly. Clinically 88.4% of the measurements were assessed plausible, in 8.2% the displayed ICP-values were judged to be too high, in 2.1% too low. Probe drift after explanation was -0.21 mmHg/24 hrs. The mean value of the recalibrated probes in the water column corresponding to 15.8 mmHg was 14.7 +/- 1.9 mmHg. There was no correlation between neither drift nor function in the water column and the duration of the measurement. Technical complications exclusively related to the construction of the CAMINO-system like kinking of the cable, dislocation (probe pulled out) or dislocated fixation screw were too high (25.3%). PMID- 9416353 TI - Trophoblast cell-mediated modifications to uterine spiral arteries during early gestation in the macaque. AB - A specialized subset of invasive embryonic cytotrophoblast cells gains access to maternal uterine arteries early in the gestation of higher primates. These cells continue to migrate extensively within the lumina of spiral arteries, converting them to the highly modified uteroplacental arteries of pregnancy. Although trophoblast cell-mediated modifications are considered critical to the progress of normal pregnancy, few studies have addressed the cellular interactions between maternal arteries and embryonic cells in situ. Macaque placentas and endometrial tissues were collected from 12 animals from day 14 of gestation (blastocyst implantation begins on day 9) to day 49. Standard indirect immunoperoxidase methods were used to identify matrix metalloproteinases (MMP-1, MMP-3, MMP-9), cathepsin B, cathepsin D, platelet-endothelial cell adhesion molecule, cytokeratins, smooth muscle actin, CD68, and factor VIII-related antigen. Cytotrophoblast cells were located deep within spiral arteries in each of the specimens examined. In some examples tightly packed clusters of cytotrophoblast occluded the lumina of invaded arteries. Initial penetration of arterial tunica intima was revealed by discontinuities in the staining pattern for factor VIII and cytotrophoblast intrusion was indicated by cytokeratin staining of the trophoblast cells. Continued cytotrophoblast intrusion into the tunica media was apparent by gaps in the smooth muscle. MMP-1, MMP-3, and MMP-9 were localized within intraluminal and intramural cytotrophoblast. By contrast, neither cathepsin B nor cathepsin D were present, although both were seen in uterine macrophages and stromal cells. Upon reaching the surrounding uterine stroma the cytotrophoblast cells ceased migration. As cytotrophoblast accumulated in the arterial wall the vascular lumen expanded. Evidence of cell death was rarely encountered in associated maternal or embryonic tissues. We conclude that intra arterial cytotrophoblast cells express several proteinases with substrate specificities sufficient to permit independent remodeling of the extracellular matrix comprising uterine artery walls. The remodeling of the arteries, which involves extensive displacement of maternal endothelium and smooth muscle, in addition to degradation and synthesis of extracellular matrix, is accomplished with little evidence of cell death or loss of the integrity of the arteries. This process provides an interesting example of cooperation between different types of interacting tissues from genetically distinct individuals. PMID- 9416354 TI - Alkaline phosphatase distribution in rat endometrial epithelium during early pregnancy: a scanning electron-microscopic study. AB - The participation of apical membranes of uterine epithelial cells in the process of blastocyst adhesion makes them an interesting object in the study of changes occurring during early pregnancy. In the study of these changes alkaline phosphatase (AIP), a typical brush border enzyme, was chosen for demonstration with the scanning electron microscope (SEM) by means of a backscatter detector. Thus the temporal and spatial pattern of enzyme activity on the uterine luminal surface was made visible with lead salt procedures. AIP activity was shown to be located on apical membranes and microvilli of endometrial epithelial cells with high activity on day 2 of pregnancy decreasing to virtually no activity on day 5. This decrease in overall AIP activity was shown to be asymmetrical with respect to the uterine cavity. It begins on the antimesometrial half of the uterine lining on day 2. A distribution pattern demarcating a presumptive implantation site along the uterine horn was not found. However, on day 5 of pregnancy, a characteristic pattern of surface folds was found, dividing the uterine horn into 'implantation segments'. In addition, SEM investigation revealed a marked variation of AIP activity from one individual cell to the next on day 2 of pregnancy resulting in a mosaic-like pattern. This pattern is lost with the decrease of AIP activity on day 5. Thus heterogeneity of uterine epithelial cells in AIP activity is apparently a feature of nonreceptive epithelium in contrast to the homogeneous epithelium on day 5. It is proposed that epithelial cell homogeneity could be a marker for uterine receptivity. PMID- 9416355 TI - 11.5-day rat embryos cultured in vitro after introduction of immunoglobulin G into the vitelline circulation. AB - The fate of rat immunoglobulin G (IgG) in the 11.5-day-rat conceptus cultured in vitro has been studied utilizing the intravitelline cannulation technique. When IgG bound to colloidal gold was introduced into the vitelline circulation, gold particles were detected on the luminal surface of embryonic endothelial cells, in both coated pits and vesicles and in various portions of the vacuolar system of the embryonic endothelial cell. By means of the radiolabeled macromolecule, it has been demonstrated that the internalized IgG was not degraded. In comparison, digested products of radiolabeled bovine serum albumin (BSA) were detected in culture media after the macromolecule was introduced into the conceptus. It was therefore concluded that the 11.5-day rat embryo captures IgG probably by receptor-mediated endocytosis and does not degrade the macromolecule, indicating that IgG is not routed to the lysosomal compartment of the endothelial cell even though the embryo has the capacity to digest BSA. It appears therefore that the embryo is endowed with the capacity to handle the IgG macromolecule well before the macromolecule is introduced into it for passive immunity. PMID- 9416356 TI - Distribution and ultrastructure of the stomata connecting the pleural cavity with lymphatics in the rat costal pleura. AB - We investigated the detailed distribution and ultrastructure of the stomata connecting the pleural cavity and the lymphatics in the rat costal pleura by scanning electron, transmission electron and light microscopy. The mesothelial cells lining the costal pleura appeared as both flattened and thick cell bodies. The thick cells possessed more rough endoplasmic reticula, Golgi complexes, mitochondria, and free ribosomes than the flattened cells. The thick cells were distributed in the intercostal regions each cephalic to the junction of the costal cartilage and bone, and in the band-like regions along the cephalic and caudal sides of each rib in the lateral and dorsal thoracic walls. In the regions lined with thick cells, there were stomata [12.9 +/- 10.3 microns2 (mean +/- SD) in area] consisting of prolongations of thick mesothelial cells and funnel-like projections of lymphatic endothelial cells that came up along the rims of the pores (5.9 +/- 3.2 microns2 in average area) in the submesothelial collagen fiber network. At the stomata, the basal lamina of the mesothelium was continuous with that of the endothelium. The mesothelial cells forming the stomata were mostly in close contact with the endothelial cells, but some gaps also existed between them. Valve-like endothelial flaps were frequently observed wherever endothelial cells constituting the stomata merged into the submesothelial lymphatics. Also present were lymphatic bulges that were either in close contact with the base of the thick mesothelial cells or exposed through the mesothelial pores. The lymphatic network was especially well developed in the submesothelial layer at and around the thick-cell regions. The initial lymphatics drained into the intercostal collecting lymphatics, which in turn led into either the parasternal or paravertebral lymphatic trunk. Our results suggest that the stomata play a major role in absorbing fluids and particulates in the pleural cavity. The thick mesothelial cells appear to secrete chemotactic substances to the endothelial cells. Understanding the heterogeneous distribution of the stomata could prove to be important clinically in inflammatory diseases and tumors in the chest. PMID- 9416357 TI - Asymmetry of normal mandibular condylar shape. AB - Morphological studies of the facial skeleton in human beings are usually made from radiographs (frontal and lateral projections and orthopantomographs). The conventional linear and angular measurements provide quantitative information only about size, and fail to define the shape and form of the skeletal features and their variations. Mathematical methods such as the Fourier series allow a correct quantitative analysis of the shape and its variations. The outlines of the mandibular condyles in the orthopantomographs of 20 men and 20 women (mean age 29 years) were traced and digitized. All subjects had a good dentition, no temporomandibular joint problems, and were referred to a dental surgery for periodontal problems. A Fourier analysis of the outlines was performed. Fourier coefficients and reconstructed outlines were compared to analyze the condylar symmetry of shape on an intra- and intersubject basis. A significant condylar asymmetry for shape as distinct from size was found on an intrasubject basis, i.e. the left and right condyles of a single individual had a different shape with a large interindividual variability. Conversely, the mean condyle shape of the male and female groups was symmetric. PMID- 9416358 TI - Lumbosacral plexus: a histological study. AB - Two hundred and forty histological paraffin sections were obtained from the midportion of the spinal nerve roots of 20 lumbosacral plexus (from L4 to S3) bilaterally. All microscopic images were digitized using the NIH image software with a Nikon microscope and Sony videocamera. The total, fascicular as well as epineurial cross-sectional areas of the nerve roots in the lumbosacral plexus were determined. The total cross-sectional area of the lumbosacral trunk (LST) was the largest (28.56 +/- 12.28 mm2) followed by the S1 (21.97 +/- 11.22 mm2) and L5 (21.00 +/- 8.79 mm2) nerve roots. The total cross-sectional areas of the L4 (6.93 +/- 3.32 mm2), S2 (13.93 +/- 5.86 mm2) and S3 (6.03 +/- 3.74 mm2) were significantly lower. Statistical differences were found among all absolute values at different levels (p < 0.0001) with the exception of the levels between L4 and S3, L5 and S1, and LST and S1 (p > 0.05). The total areas occupied by the fascicles in L5 (7.78 +/- 3.26 mm2), LST (9.97 +/- 4.01 mm2) and S1 (8.55 +/- 3.27 mm2) nerve roots were greater than those in L4 (2.96 +/- 1.50 mm2), S2 (5.56 +/- 2.34 mm2) and S3 (2.28 +/- 1.14 mm2). However, the percentages of the total cross-sectional areas occupied by the fascicles in the L5 nerve root (38%) and LST (36.4%) were smaller compared to other nerve roots (44.9% at L4, 40.5% at S1, 40.8% at S2 and 41.6% at S3). The cross-sectional areas and percentages of the epineurium were greater in L5 (13.22 +/- 6.48 mm2, 62.0%), LST (18.58 +/- 9.31 mm2, 63.6%) and S1 (13.42 +/- 8.88 mm2, 59.5%) roots. The L5 (12.1 +/- 5.0), LST (27.5 +/- 11.4) and S1 (15.0 +/- 7.3) roots contained more fascicles than L4 (6.3 +/- 3.3), S2 (9.5 +/- 4.1) and S3 (7.1 +/- 2.7). Statistical differences were found among all absolute values at different levels (p < 0.0001) with the exception of the levels between L4 and S3, L5 and S1 and LST and S1 (p > 0.05). No statistical differences were found for percentages among different levels (p > 0.05) with the exception of the levels between L4 and L5, L4 and LST, LST and S1, LST and S2, and LST and S3 (p < or = 0.05). The histological structure of nerve roots of the lumbosacral plexus is identical to that of the peripheral nerve. The midportions of L5 and S1 roots and the LST have a relatively higher epineurial content. These nerve roots also have a greater number of the fasicles, but the total cross-sectional area occupied by the fascicles is less than in L4, S2 and S3 nerve roots. PMID- 9416359 TI - A histochemical, morphometric and ultrastructural study of gastrocnemius and soleus muscle fiber type composition in male and female rats. AB - The fiber type composition of gastrocnemius and soleus muscles of adult male and female rats was examined by histochemical, morphometric and ultrastructural methods. Six male and 6 female, 3.5-month-old rats were used in the study. The fiber types were determined using the adenosine triphosphatase (ATPase) and succinate dehydrogenase (SDHase) staining techniques. The number of fibers of different types and the diameters of these two muscles were examined by morphometric methods. Gastrocnemius muscles were also examined by transmission electron microscopy. Z-line thickness, actin length, sarcomere length and M-line thickness of the fiber types were compared at the ultrastructural level. No significant differences were observed between the diameters of the gastrocnemius muscle (p > 0.05) of male and female rats although the male animals were significantly larger than the females (p > 0.05). In males, type I fibers of the gastrocnemius were larger than in females (p < 0.05). Region II of the gastrocnemius muscles of females contained more type IIA and type IIB fibers than in males. Sarcomere lengths were also longer in females (p < 0.05). The composition of fiber types of gastrocnemius and soleus muscles may be sex dependent. PMID- 9416360 TI - Morphological analysis of the human tibialis anterior and medial gastrocnemius muscles. AB - The morphology and nerve innervation patterns of eight tibialis anterior (TA) and medial gastrocnemius (MG) muscles from human cadavers were examined. TA has three distinct partitions, two of which lie posteriorly. The anterior aspect of the muscle has one partition (head A) whose fibers are oriented longitudinally. The posterior partitions possess fibers that course longitudinally (B head) and obliquely (C head). The posterior longitudinal fibers and the anterior longitudinal fibers are divided by an aponeurosis that is continuous with the tendon of insertion. The proximal muscle fibers insert into this aponeurosis and the distal fibers insert into the tendon. The MG is a unipennate muscle. Both TA and MG muscles show variations in innervation patterns among specimens. In light of these findings, partitioning of TA may be based on its architecture. PMID- 9416361 TI - The influence of forearm, hand and thumb positions on extensor carpi ulnaris and abductor pollicis longus activity. AB - The objective of this study was to investigate the influence of forearm, hand and thumb positions on electromyography (EMG) activity of the abductor pollicis longus (APL) and the extensor carpi ulnaris (ECU) muscles. A second objective was to study the role of these muscles in stabilizing the wrist during movements of the thumb. This knowledge is important in clinical assessment and reconstructive surgery. At a constant force of 5 N for the thumb and 20 N for the hand, EMG activity was recorded with intramuscular wire electrodes in isometric and dynamic contractions in different positions and movements in the thumb and the hand. EMG activity of the right hand of 7 subjects was scaled relative to the mean EMG value at maximum voluntary isometric contraction to compare relative muscle activity in the tests. The results show that the position of the forearm does influence activity of the APL, but not of the ECU. The deep part of the APL shows differences in EMG activity between the positions of the forearm in dorsal flexion of the hand and the superficial part of the APL in palmar flexion of the hand. The ECU and the deep part of the APL are very active during movements and isometric contractions of the thumb. It is suggested that these muscles are necessary to stabilize the wrist during movements of the thumb to prevent undesired movements of the hand and the forearm. PMID- 9416362 TI - HIV/AIDS in Africa: the second decade and beyond. PMID- 9416363 TI - HIV/AIDS epidemics in sub-Saharan Africa: dynamism, diversity and discrete declines? PMID- 9416364 TI - The making of HIV epidemics: what are the driving forces? PMID- 9416365 TI - Virology of HIV-1 and HIV-2: implications for Africa. PMID- 9416366 TI - Natural history and spectrum of disease in adults with HIV/AIDS in Africa. AB - Progression from seroconversion to the development of AIDS in Africa may be shorter than in industrialized countries, but there are insufficient data to be certain. Although the data are not always directly comparable, survival after an AIDS diagnosis appears to be substantially shorter in African countries and this may be partly because of later diagnosis of AIDS in Africa, but may also be because of environmental factors such as increased exposure to pathogens of high virulence and lack of access to care. Tuberculosis and bacterial infections are the most important causes of morbidity and mortality among hospitalized patients. Bacteraemia is frequent, particularly due to non-typhoid salmonellae and S. pneumoniae. Cryptosporidia and I. belli are the most frequently isolated pathogens in patients with diarrhoea; non-typhoid salmonellae and Shigella species are also commonly isolated when stool cultures are performed. Cerebral toxoplasmosis, and meningitis due to Cryptococcus, tuberculosis and bacterial pathogens are the most frequent neurological infections and cognitive changes are frequently identified when specifically looked for. Infections with atypical mycobacteria and Pneumocystis carinii are rare, as is CMV retinitis. In women, HIV infection is associated with cervical human papillomavirus and with SIL, although there is currently no evidence for an association with invasive cervical cancer. Individuals infected with HIV-2 progress to AIDS and to death more slowly than those infected with HIV-1, but seem to experience the same spectrum of opportunistic disease when they reach the stage of advanced disease. The limited data available suggest that HIV-infected individuals in Africa develop opportunistic disease at broadly the same level of immunosuppression as do individuals in industrialized countries, but death occurs at a higher range of CD4 counts, although still in the range consistent with advanced disease. Data are still lacking concerning the aetiology of common clinical presentations of HIV disease and the relative frequencies of specific opportunistic diseases in different regions, particularly from southern Africa. Tuberculosis is the single most important HIV-related opportunistic infection in African countries, but diagnosis, particularly of extrapulmonary disease, remains difficult. The lack of laboratory facilities makes the diagnosis of bacterial infections difficult in many parts of the continent and, since this situation is unlikely to change in the near future, clinical algorithms for syndromic management need to be evaluated. More information is needed about gynaecological disease in HIV infected women. The most important research questions concern the development and evaluation of cost-effective regimes for prophylaxis and treatment of opportunistic disease in order to prolong healthy life in HIV-infected individuals. PMID- 9416367 TI - The demographic and economic impact of AIDS in Africa. PMID- 9416368 TI - Prevention of sexual transmission of HIV in sub-Saharan Africa: lessons learned. PMID- 9416369 TI - Prevention of mother-to-child transmission of HIV-1 in Africa. PMID- 9416370 TI - Prevention of blood-borne transmission of HIV. PMID- 9416371 TI - The challenge of providing effective care for HIV/AIDS in Africa. PMID- 9416372 TI - Antiretroviral treatment in Africa. PMID- 9416374 TI - Care of children with HIV infection and AIDS in Africa. PMID- 9416373 TI - Tuberculosis and HIV: current status in Africa. PMID- 9416375 TI - Models of care for patients with HIV/AIDS. PMID- 9416376 TI - Government responses to HIV/AIDS in Africa: what have we learnt? AB - While we should not lose sight of the development of vaccines and cures, more immediate priorities include the implementation of effective STD control. The syndromic management approach developed in Zimbabwe to overcome laboratory constraints is a cost-effective way of managing STD. Of urgency is the integration of STD services into primary health-care services, appropriate training of staff, adequate provision and control over drugs and condoms, and incorporation of traditional healers and community-based education on STD. A second area of priority is the strengthening of the interaction between prevention, care and support activities, which act in synergy. Effective prevention and care require easy access to testing facilities with pre- and post test counselling, appropriate structures and services to provide affordable and sustained care and support to those found to be infected with HIV, provision of drugs for the treatment of opportunistic infections, and the creation of a social environment and a legislation which protect against any form of discrimination the rights of people living with HIV, their sexual partners and their families. PMID- 9416377 TI - Non-governmental organizations and community responses to HIV/AIDS and the role of HIV-positive persons in prevention and care. PMID- 9416378 TI - A human rights perspective on HIV/AIDS in sub-Saharan Africa. PMID- 9416379 TI - Advanced HIV infection treated with zidovudine monotherapy: lifetime values of absolute cost-effectiveness as a pharmacoeconomic reference for future studies evaluating antiretroviral combination treatments. The Osservatorio SIFO sui Farmaci. AB - OBJECTIVE: This study was undertaken to evaluate the cost and the effectiveness of zidovudine monotherapy in patients with advanced HIV infection and to derive preliminary data on the cost-effectiveness of the triple treatment with saquinavir plus zalcitabine plus zidovudine compared with zidovudine alone. DESIGN: We used a combined method of survival analysis utilizing both the quality adjusted time without symptoms or toxicity (Q-TWIST) method and the Gompertz approach. This combined method was applied to assess the absolute cost effectiveness and cost-utility ratios of zidovudine monotherapy and to perform a preliminary incremental cost-effectiveness comparison of saquinavir plus zalcitabine plus zidovudine versus zidovudine alone. The clinical material used in our study was derived from two reports on the treatment of advanced HIV infection. Data of lifetime costs of HIV infection were obtained from published information. RESULTS: In patients with advanced HIV infection treated with zidovudine monotherapy, lifetime survival was 252.1 discounted person-years per 100 patients. Using an average lifetime cost of $93,000 (discounted) per individual, the absolute ratio of cost-effectiveness for zidovudine monotherapy was $36,980 per life-year, while the absolute cost-utility ratio was $47,112 per quality-adjusted life-year. In the comparative analysis of saquinavir plus zalcitabine plus zidovudine versus zidovudine alone, our calculations showed that the administration of the triple treatment can have an "average" cost effectiveness, provided that mean lifetime survival per patient (discounted) is improved to at least 3.68 years (with an average survival gain of at least 14 mo per patient). CONCLUSIONS: The values of absolute cost-effectiveness and cost utility ratios for zidovudine monotherapy are a useful reference point for further pharmacoeconomic studies in the area of antiretroviral drugs. PMID- 9416380 TI - Reevaluation of a weight-based heparin dosing nomogram: is institution-specific modification necessary? AB - OBJECTIVE: To compare a heparin dosing nomogram using an initial infusion rate of 18 units/kg/h with physician-directed heparin prescribing and with a modified version of the nomogram adjusted for institution-specific data. METHODS: During consecutive phases of this cohort study, patients' intravenous heparin therapies were initiated and adjusted by using one of the following three methods: (1) physician-directed dosing, (2) a body weight-based dosing nomogram with an initial infusion rate of 18 units/kg/h, and (3) a body weight-based dosing nomogram with an initial infusion rate determined by the median dose of heparin (in units/kg/h) required to achieve therapeutic activated partial thromboplastin times (aPTTs) during the first two phases. The time required to achieve therapeutic aPTTs as well as the percentage of initial aPTTs in the therapeutic range were compared for the three phases. RESULTS: The heparin dosing nomogram in which the initial infusion rate was adjusted for our individual institution resulted in a statistically shorter median time until aPTTs were in the therapeutic range than did either the physician-directed dosing or unmodified nomogram groups (6.1 h in the modified nomogram group, 10.5 h in the physician directed group, 21.5 h in the unmodified nomogram group; p < 0.05 for all differences). Use of the institution-specific nomogram resulted in the greatest percentage of initial aPTTs in the therapeutic range (84% in the 13 units/kg/h nomogram group vs. 47% in the physician-directed group and 18% in the 18 units/kg/h nomogram group; p < 0.05 for all differences). CONCLUSIONS: Use of a heparin dosing nomogram with an initial infusion rate of 18 units/kg/h resulted in prolongation of the time to reach therapeutic aPTTs. By modifying the nomogram for use at an individual institution, we reduced the time to achieve therapeutic range of aPTTs while still reducing the likelihood of excessive anticoagulation of patients. PMID- 9416381 TI - Reduced tetracycline bioavailability caused by magnesium aluminum silicate in liquid formulations of bismuth subsalicylate. AB - RATIONALE: Bismuth subsalicylate, tetracycline hydrochloride, and metronidazole are widely used in combination for the treatment of Helicobacter pylori infections. As a result, there is renewed interest in the interaction between tetracycline and bismuth subsalicylate. OBJECTIVE: To determine whether the observed decrease in tetracycline bioavailability is due to the active drug bismuth subsalicylate via complexation, or to magnesium aluminum silicate (Veegum), an inactive excipient present only in the liquid formulation of bismuth subsalicylate, which might adsorb the tetracycline, rendering it unavailable for systemic absorption. METHODS: Eleven healthy volunteers participated in a randomized three-period, three-treatment complete crossover study with a 7-day washout interval between treatments. After an overnight fast, subjects received a 500-mg capsule of tetracycline hydrochloride with either tap water, 30 mL of bismuth subsalicylate (525 mg) liquid containing Veegum (Pepto-Bismol), or 30 mL of a specially formulated bismuth subsalicylate (525 mg) liquid without Veegum. Blood was collected for 24 hours after each dose of tetracycline. Serum was assayed for tetracycline concentration by HPLC. In addition, standard in vitro ultraviolet spectrophotometric methods were used to determine the capacity for complexation of bismuth with tetracycline and for adsorption of tetracycline to Veegum. RESULTS: Compared with the reference treatment of tetracycline hydrochloride with water, the liquid formulation of bismuth subsalicylate containing Veegum decreased the maximum serum concentration (Cmax) of tetracycline by 21% and the serum tetracycline AUC by 27% (p < 0.001). The bismuth subsalicylate formulation without Veegum resulted in decreases in Cmax and AUC of 11% and 13%, respectively (p > 0.05 vs. tetracycline hydrochloride with water). Multiple linear regression analysis of the spectral absorbance data demonstrated a calculated recovery of tetracycline of 100.9% and, therefore, a lack of in vitro complexation with bismuth. At pH 1.2, the amount of tetracycline adsorbed to Veegum ranged from 91.5% to 97.2% over the concentration range of 0.25 to 2 mg/mL. At pH 7.0, the values ranged from 82.9% to 83.9% over the concentration range of 0.25 to 1 mg/mL. CONCLUSIONS: In vitro and in vivo data from this study indicate that Veegum, a suspending agent, and not the active agent bismuth subsalicylate, is the primary ingredient in liquid formulations of bismuth subsalicylate responsible for a decrease in tetracycline bioavailability. In addition, the mechanism of interaction is not likely due to complexation between tetracycline and bismuth subsalicylate, as previously postulated, but rather is caused by adsorption of tetracycline to the excipient Veegum, which is present only in the liquid formulation of bismuth subsalicylate. The clinical relevance of this interaction has not been determined. PMID- 9416382 TI - Compatibility and stability of paclitaxel combined with cisplatin and with carboplatin in infusion solutions. AB - OBJECTIVE: To evaluate the physical compatibility and chemical stability of paclitaxel at concentrations of 0.3 and 1.2 mg/mL with cisplatin 0.2 mg/mL in NaCl 0.9% injection and with carboplatin 2 mg/mL in NaCl 0.9% injection and dextrose 5% injection over 7 days at 4, 23, and 32 degrees C. DESIGN: The test samples were prepared in polyolefin bags of the infusion solutions at the required drug concentrations. Evaluations were performed initially and after 4 hours, and 1, 3, 5, and 7 days of storage at temperatures of 4, 23, and 32 degrees C for physical and chemical stability. Physical stability was assessed by using visual observation in normal light and using a high-intensity monodirectional light beam. In addition, turbidity and particle content were measured electronically. Chemical stability of the three drugs was evaluated by using three stability-indicating HPLC analytical techniques. RESULTS: All samples were physically stable through 1 day. However, microcrystalline precipitation of paclitaxel occurred in 3 days in some samples and within 5 days in all samples. Paclitaxel concentrations remained above 90% in all samples throughout the study. Cisplatin admixtures exhibited paclitaxel concentration-dependent decomposition with cisplatin losses of approximately 5-8% in 4 hours and approximately 20% in 1 day at 23 and 32 degrees C in the paclitaxel 1.2 mg/mL admixtures. With paclitaxel 0.3 mg/mL in the admixtures, cisplatin losses were about 10% in 7 days at these temperatures. Carboplatin in admixtures with both concentrations of paclitaxel was stable for 7 days at 4 degrees C, but sustained losses of about 10% and 12% in 3 days at 23 and 32 degrees C, respectively. CONCLUSIONS: Admixtures of paclitaxel 0.3 and 1.2 mg/mL with cisplatin and carboplatin are limited in their utility time by both paclitaxel microcrystalline precipitation and decomposition of cisplatin and carboplatin. The admixture of paclitaxel 1.2 mg/mL with cisplatin 0.2 mg/mL in NaCl 0.9% injection exhibits unacceptable cisplatin loss in 24 hours. All other combinations were physically and chemically stable for at least 24 hours at 4, 23, and 32 degrees C. PMID- 9416383 TI - Paclitaxel-induced acute bilateral pneumonitis. AB - OBJECTIVE: To report three cases of paclitaxel-induced acute bilateral pneumonitis, as well as to ascertain its incidence and outcome. CASE SUMMARIES: A total of 239 patients with a variety of underlying malignancies received 528 courses of paclitaxel-containing chemotherapy. Paclitaxel 200 mg/m2 was infused over 3 hours with standard premedication. Three patients developed bilateral interstitial infiltrates either during or within 6 hours of the administration of paclitaxel. Symptoms included a nonproductive cough, dyspnea, and sudden arterial oxygen desaturation. Response to parenteral corticosteroids was dramatic and reversed the process in all 3 patients. DISCUSSION: Paclitaxel-induced acute bilateral pneumonitis appears to be a rare adverse reaction. It may either be a direct toxic effect of the chemotherapeutic agent or an adverse effect of its Cremophor EL diluent. Although the exact pathophysiology is unclear, a variety of immune and nonimmune mechanisms have been postulated, including hypersensitivity reactions, release of cytokines from macrophages, and the possible role of prior thoracic irradiation. CONCLUSIONS: Acute bilateral pneumonitis occurs in less than 1% of individuals receiving 3-hour infusions of paclitaxel, and responds dramatically to parenteral corticosteroid therapy. PMID- 9416384 TI - Ofloxacin-induced seizure. AB - OBJECTIVE: To describe a possible case of ofloxacin-induced generalized tonic clonic seizure. Although the etiology is unknown, ofloxacin most likely precipitated this patient's seizure threshold because of sepsis or secondary to drug accumulation due to the patient's compromised renal function. CASE SUMMARY: A 69-year-old white woman with non-small-cell lung cancer and a history of central nervous system metastatic disease treated with radiation therapy presented to the emergency department with symptoms of urosepsis. Because of multiple drug allergies she was started on ofloxacin (hospital formulary quinolone). After 4 days of therapy she developed a generalized tonic-clonic seizure. A computed tomography scan of the head with and without contrast was negative. The ofloxacin was discontinued and aztreonam therapy was started. Phenytoin therapy was instituted and, despite serum concentrations below the conventional therapeutic range, there was no recurrence of seizure. Subsequent discontinuation of phenytoin did not result in a seizure for this patient. DISCUSSION: Seizures induced by the fluoroquinolones are uncommon. The histopathologic features of this phenomenon are currently unknown. In this patient, imaging studies were negative for structural defects, ruling out metastasis as the cause of the seizure. Therefore, an investigation of drug related causes ensued. The most likely offending agent was ofloxacin. Ofloxacin has been reported in the literature as a cause of seizures in patients with compromised renal function. CONCLUSIONS: This case and other reports indicate that fluoroquinolones, including ofloxacin, may contribute to seizure development in patients with or without a history of epilepsy. Fluoroquinolone therapy should be used with caution in patients with risk factors for the development of drug induced seizures. PMID- 9416385 TI - Fluoxetine-associated stomatitis. AB - OBJECTIVE: To describe two cases of stomatitis related to fluoxetine given for the treatment of depression that were detected in the hospital emergency department. DATA SYNTHESIS: Two women developed stomatitis after the intake of fluoxetine for the treatment of depression. One of the patients had six recurrent episodes of stomatitis without suspecting an association with fluoxetine. No other drugs were administered during these episodes. The second patient was treated concurrently with fluoxetine and bentazepam. In both patients the lesion improved upon discontinuation of fluoxetine, even though the second patient continued to take a different benzodiazepine. DISCUSSION: Stomatitis related to fluoxetine has not been previously reported in clinical trials or in the literature. According to the causal algorithm used by the Spanish Drug Surveillance Schemes, the first case constituted a defined adverse reaction and the second was probable. CONCLUSIONS: Our observations suggest that fluoxetine may be considered as a probable cause of stomatitis. The reporting of isolated cases of adverse drug reactions (ADRs) makes it possible to define the toxicity profile of recently marketed drugs such as selective serotonin-reuptake inhibitors, including fluoxetine. Emphasis is placed on the potential role played by emergency departments in detecting ADRs. PMID- 9416386 TI - Extrapyramidal reactions and the selective serotonin-reuptake inhibitors. AB - OBJECTIVE: To review the known published reports of extrapyramidal reactions (EPRs) associated with the use of selective serotonin-reuptake inhibitors (SSRIs). DATA SOURCES: Information was selected from a MEDLINE search (January 1990 to January 1996) of English-language medical literature. Manual searches of pertinent journal article bibliographies were also performed. DATA EXTRACTION: Appropriate information from all reports obtained was included, with specific attention directed toward patient age, gender, primary psychiatric diagnosis, total daily SSRI dosage, dosage escalation strategy, and concurrent psychotropic medications. DATA SYNTHESIS: Reports of EPRs associated with SSRI use have been accumulating in the medical literature for several years. More commonly associated with high-potency antipsychotics, EPRs can have an adverse impact on medication compliance and hospital readmissions. The proposed hypothesis for EPRs occurring with SSRI use involves serotonin's inhibitory actions on extrapyramidal dopamine activity. Other possible contributing factors include pharmacokinetic interactions or drug-disease interactions. EPRs may include dystonias, dyskinesias, akathisia, parkinsonism, exacerbations of Parkinson's disease, and possibly the neuroleptic malignant syndrome. The majority of SSRI-related reactions appear to occur within the first month of treatment. Information from available case reports does not strongly support any consistent risk factor, although some worth considering may include total SSRI daily dose, rapid dose escalation strategies, increased age, female gender, concurrent psychotropics known to also precipitate EPRs, and concurrent disease states such as Parkinson's disease. Since SSRI-related EPRs have occurred in different situations with different possible contributing factors, clinical pharmacy practitioners and other healthcare providers should remain aware of these reactions and carefully consider educating and monitoring their patients accordingly. CONCLUSIONS: The use of SSRIs may be associated with the development of EPRs; therefore, appropriate monitoring should be considered for patients so that optimal pharmaceutical care may be provided. PMID- 9416387 TI - Introduction to coronary artery stents and their pharmacotherapeutic management. AB - OBJECTIVE: To provide an introduction to coronary artery stents and their pharmacologic management, including anticoagulant therapy and newer antiplatelet regimens. DATA SOURCES: A MEDLINE and current journal search of relevant articles that evaluated coronary stent success rates and anticoagulation or antiplatelet regimens. STUDY SELECTION: Data from the use of primarily the Palmaz-Schatz stent were included. Studies using vitamin K antagonists that are not commercially available in the US were excluded unless they compared an antiplatelet regimen with anticoagulation using the international normalized ratio (INR). DATA SYNTHESIS: Limitations with percutaneous transluminal coronary angioplasty (PTCA), such as ischemic complications and restenosis, have led to the advent of intracoronary stenting. However, the placement of a stent within the coronary artery lumen is associated with a risk of thrombotic events. Despite current postprocedural anticoagulation and antiplatelet regimens, thrombosis occurs at rates ranging from 0.6% to 21%. When anticoagulation is deemed appropriate, it should be used for 1-2 months and the INR should be maintained between 2 and 3.5. Anticoagulation appears to have no effect on the development of restenosis, but has been shown to cause significant hemorrhagic events in 5-13.5% of patients. Newer data continue to define the subsets of patients who may be managed with antiplatelet agents alone. Combinations of aspirin and ticlopidine or aspirin alone may be used to manage patients who fulfill the following criteria: optimal stent placement, high-pressure inflation, and adequate coronary size. CONCLUSIONS: Coronary artery stenting is a novel approach for the management of coronary artery disease, but is associated with the complication of stent thrombosis. Anticoagulation reduces the risk of stent thrombosis, but is associated with bleeding risk. Selected patients may be successfully managed with antiplatelet agents only. More data are needed to better define the optimal antithrombotic regimen. PMID- 9416388 TI - Current perspective on the use of angiotensin-converting enzyme inhibitors in the management of coronary (atherosclerotic) artery disease. AB - OBJECTIVE: To review the pathophysiology of atherosclerosis, the role of the renin-angiotensin system in atherogenesis, and studies supporting the potential beneficial effects of angiotensin-converting enzyme (ACE) inhibitors in reducing cardiovascular events with long-term use. BACKGROUND: Through its action in converting angiotensin I to angiotensin II and by degrading bradykinin, local tissue ACE exerts many effects that can contribute to the development of atherosclerosis. Therefore, the use of ACE inhibitors can possibly result in antiatherogenic effects. Possible mechanisms for antiatherogenic effects of ACE inhibitors include: (1) reduction of blood pressure; (2) antiproliferative and antimigratory effects on vascular smooth muscle cells, neutrophils, and monocytes; (3) restoration of endothelial function; (4) stabilization of fatty plaque by preventing vasoconstriction; (5) antiplatelet effects; and (6) enhancement of endogenous fibrinolysis. DATA SOURCES: English-language clinical studies, abstracts, and review articles pertaining to the use of ACE inhibitors and atherosclerosis. STUDY SELECTION AND DATA EXTRACTION: Relevant human studies examining the role of ACE inhibitors and atherosclerosis. DATA SYNTHESIS: Studies evaluating the possible beneficial effects of ACE inhibitors in the development of atherosclerosis are reviewed and critiqued. Design of ongoing studies with clinical and surrogate end points are discussed. CONCLUSIONS: Based on current published studies, recommendations are made regarding the use of ACE inhibitors in atherosclerosis. Therapeutic monitoring parameters for efficacy and adverse effects are also reviewed. PMID- 9416389 TI - Tetanus: pathophysiology and management. AB - OBJECTIVE: To review the epidemiology, pathophysiology, clinical manifestations, diagnosis, and management of tetanus and its complications. DATA SOURCES: MEDLINE and Iowa Drug Information Services databases were searched for literature pertaining to tetanus. Additional literature was obtained from the references of selected articles identified in the search. Information from all articles was considered for inclusion in the manuscript. STUDY SELECTION AND DATA EXTRACTION: Articles selected were those considered by the authors to assist in providing the reader an understanding of the epidemiology, pathophysiology, clinical manifestations, diagnosis, and management of tetanus. DATA SYNTHESIS: While the number of tetanus cases has decreased markedly since data reporting for the disease began in 1947, mortality among those who acquire the disease remains high. Elderly patients are particularly susceptible to tetanus and its complications. Prevention of tetanus focuses on primary immunization and scheduled boosters. Management of tetanus involves initial stabilization of the patient and protection of the airway, prevention of tetanospasmin absorption by administration of human tetanus immune globulin 3000-6000 IU, and eradication of Clostridium tetani with antimicrobial therapy (metronidazole 500 mg q8h). Supportive measures include the administration of neuromuscular blocking agents such as pancuronium in patients requiring artificial ventilation, as well as benzodiazepines (midazolam 5-15 mg/h) for sedation and muscle relaxation. Autonomic dysfunction should be managed with beta-adrenergic blockers such as propranolol or labetalol. CONCLUSIONS: Despite the relative infrequency of tetanus cases, mortality among untreated patients remains significantly high. Clinicians should become knowledgeable in the pathophysiology, clinical manifestations, and management of this potentially fatal disease. PMID- 9416390 TI - Routine monitoring of gentamicin serum concentrations in pediatric patients with normal renal function is unnecessary. AB - OBJECTIVE: Due to increasing demands for cost containment within the healthcare system, we evaluated the need for routine gentamicin concentrations (i.e., peak and trough with third dose). DESIGN: Single-institution study performed concurrently with hospitalization. SETTING: A 225-bed pediatric teaching hospital. PARTICIPANTS: The study population consisted of 150 hospitalized pediatric patients (53% medicine, 47% surgical patients) from 3 months to 15 years old with normal serum creatinine. OUTCOME MEASURES: If the administered dose produced diagnoses-appropriate peak concentrations of at least 4 micrograms/mL or 5 micrograms/mL in bacteremia/septicemia and at least 6 micrograms/mL or 8 micrograms/mL in patients with pneumonia if trough serum gentamicin concentrations were less than 2 micrograms/mL, if the patient was noted by the attending physician to be clinically responding as well as objectively having a decreased white blood cell count and was afebrile, and if there was not an increase of 0.5 mg/dL or more in serum creatinine during the course of therapy. RESULTS: Patients received a mean dose of gentamicin 2.51 +/- 0.14 mg/kg i.v. q8h, which resulted in a mean peak concentration of 6.1 +/- 1.7 micrograms/mL (range 2.4-11.7) and a mean trough concentration of 0.5 +/- 0.3 microgram/mL (range 0.1-1.8). Peak and trough concentrations were at least 4 micrograms/mL and less than 2 micrograms/mL in 96% and 100% of patients, respectively. No patient required a dosage change due to lack of clinical response. CONCLUSIONS: Our data do not support the routine monitoring of gentamicin concentrations in pediatric patients older than 3 months of age who are receiving appropriate standard doses of gentamicin and have normal renal function. PMID- 9416391 TI - Cidofovir use in acyclovir-resistant herpes infection. AB - Herpes infections continue to be prevalent, especially in immunocompromised patients. Some of these patients will develop resistant HSV infections. Therefore, it is important to explore new treatment options. Animal studies have shown cidofovir to be effective in the treatment and prevention of HSV infections. Human data are limited, with only one randomized, double-blind, placebo-controlled trial performed to date. The results from this study look promising; however, due to the small sample size, a larger clinical trial is warranted. The human data available as case reports are suboptimal in the quality of reporting time frames for resolution of lesions/symptoms and outcomes of therapy. Another problem with the case report data is that the TK status of the herpes simplex isolates was not reported. This would have helped substantiate the acyclovir resistance seen in these patients. It was evident in these case reports that acyclovir resistance can be overcome, as acyclovir-resistant strains became sensitive following cidofovir therapy. This may be because TK(+) viruses have been shown to establish latency more readily than do TK(-) viruses. This pattern suggests that alternating between acyclovir and cidofovir therapies may provide a strategy to manage the emergence of alternatively acyclovir-sensitive and resistant infections. At present, only the intravenous formulation of cidofovir is commercially available. Advantages of the intravenous formulation include weekly dosing and efficacy. Disadvantages are the complexity of administration and the adverse effect profile. The most common adverse effects with this formulation include nephrotoxicity manifested as proteinuria (12%), and increased creatinine (5%) and neutropenia (15%). Administration of probenecid and NaCl 0.9% hydration are used to reduce the incidence and severity of nephrotoxicity in patients who are receiving cidofovir. Probenecid also has toxicities, including nausea, vomiting, headache, fever, and flushing. The topical formulation of cidofovir looks promising for mucocutaneous HSV infection because it is usually undetectable in the blood following topical administration. Therefore, systemic adverse effects should be minimized. A cidofovir gel product (Forvade, Gilead Sciences) is currently being reviewed by the Food and Drug Administration for the treatment of refractory HSV. Ultimately, more controlled clinical studies are necessary to determine whether routine cidofovir use can be justified in patients with acyclovir-resistant HSV infection. PMID- 9416392 TI - Propofol and postoperative infections. AB - The package insert for propofol states in several places the importance of strict aseptic technique in the preparation and handling of the drug, and the potential for infection if these procedures are not followed. In the US, the product has been reformulated to contain disodium edetate 0.005% as a microbial growth retardant to inhibit the growth of microorganisms in the event of accidental contamination. However, this new formulation is not considered an antimicrobially preserved product under USP standards. What effect, if any, this has had on the risk of postoperative infections is not known. Strict aseptic technique is still required when handling the new formulation. When propofol is used as an anesthetic, it is recommended that the dose be prepared just prior to administration, and that the infusion be completed within 6 hours after the ampul or vial is opened. Any unused portion of propofol must be discarded at the end of the procedure or at 6 hours, whichever occurs sooner. In the ICU, if propofol is administered directly from its original container, the tubing and any unused portion must be discarded after 12 hours. If propofol is transferred to another container, administration lines and unused drug should be discarded after 6 hours. The occurrence of postoperative infections is usually thought to be related to the surgeon or the surgical procedure. However, based on the available microbiologic and clinical evidence, the use of propofol appears to be an additional risk factor. Hospital personnel involved in the preparation and handling of propofol must be educated on, and adhere to, the recommendations outlined by the manufacturer to prevent further outbreaks of infections. PMID- 9416393 TI - Vasopressin use in cardiopulmonary resuscitation. AB - While these preliminary results in humans seem promising, no large trials have been conducted to compare vasopressin and epinephrine in CPR. At this time, standard guidelines using epinephrine are still preferred. PMID- 9416394 TI - Overview of the hospital formulary systems in Hong Kong. Princess Margaret Hospital as a baseline. AB - OBJECTIVE: To investigate the popularity of formulary systems in all Hong Kong hospitals and to compare these with the newly introduced formulary system in a major government hospital, the Princess Margaret Hospital (PMH), as the baseline. DESIGN: Questionnaire and selected interviews by pharmacy students. SETTING: All hospital pharmacies in Hong Kong. PARTICIPANTS: Department managers (directors of pharmacy services) of hospital pharmacies. MAIN OUTCOME MEASURE: The popularity of the hospitals' formulary systems and their formulary decision-making strategies. Calculations of cost savings of the new formulary system in PMH and a comparison of the PMH system with the US standards were also made. RESULTS: Among 38 responding hospitals, 35 (92%) had a formulary handbook and 21 (55.3%) claimed to have a formulary system. The evaluation processes and formulary decision making procedures were found to be inadequate because basic components in drug evaluation (e.g., standardized criteria for drug evaluation) were not used regularly. However, the formulary system in PMH was found to be comparable with the US standards. Substantial cost savings were made through rejection of less cost-effective drugs by the Formulary Subcommittee in PMH. CONCLUSIONS: In general, comprehensive formulary systems are still not popular in Hong Kong. This may be due to insufficient staffing and lack of administrative and physicians' support. The new formulary system in PMH can be used as a model to develop a successful formulary system in which hospital pharmacists can prove their expertise for the benefit of both hospitals and patients in Hong Kong. PMID- 9416395 TI - Added qualifications: a new dimension in pharmacy specialty certification. PMID- 9416396 TI - Added qualifications for pharmacy specialists: another perspective. PMID- 9416397 TI - Underutilization of accepted therapies: an overlooked component of pharmaceutical care. PMID- 9416398 TI - Gentamicin monitoring in pediatric patients. PMID- 9416399 TI - Amiodarone-induced pulmonary hypersensitivity. PMID- 9416400 TI - Glossitis, stomatitis, and black tongue with lansoprazole plus clarithromycin and other antibiotics. PMID- 9416401 TI - Acute tumor lysis syndrome associated with paclitaxel. PMID- 9416402 TI - Comment: ciprofloxacin-phenytoin interaction. PMID- 9416403 TI - Comment: clinical experiences of visual disturbances with nilutamide. PMID- 9416404 TI - Update: trends in AIDS incidence--US, 1996. PMID- 9416405 TI - Comparison of positron emission tomography and computed tomography in detection of recurrent and metastatic colorectal cancer. PMID- 9416406 TI - Translational research in breast cancer angiogenesis. PMID- 9416407 TI - Detection of recurrent and metastatic colorectal cancer: comparison of positron emission tomography and computed tomography. AB - BACKGROUND: This study evaluates the clinical value of positron emission tomography (PET) with 2-[F-18] fluoro-2-deoxy-D-glucose (FDG) as compared to computed tomography (CT) in patients with suspected recurrent or metastatic colorectal cancer (CRC). METHODS: A retrospective review of the records of 58 patients who had FDG-PET for evaluation of recurrent or advanced primary CRC was performed. FDG-PET results were compared with those of CT and correlated with operative and histopathologic findings, or with clinical course and autopsy reports. RESULTS: Recurrent or advanced primary CRC was diagnosed in 40 and 11 patients, respectively. The sensitivity and specificity of FDG-PET were 91% and 100% for detecting local pelvic recurrence, and 95% and 100% for hepatic metastases. These values were superior to CT, which had sensitivity and specificity of 52% and 80% for detecting pelvic recurrence, and 74% and 85% for hepatic metastases. FDG-PET correctly identified pelvic recurrence in 19 of 21 patients; CT was negative in 6 of these patients and equivocal in 4. FDG-PET was superior to CT in detecting multiple hepatic lesions and influenced clinical management in 10 of 23 (43%) patients. CONCLUSION: FDG-PET is more sensitive than CT in the clinical assessment of patients with recurrent or metastatic CRC, and provides an accurate means of selecting appropriate treatment for these patients. PMID- 9416408 TI - TGF beta-1 regulation of VEGF production by breast cancer cells. AB - BACKGROUND: Angiogenesis is essential for tumor growth and metastasis. Vascular endothelial growth factor (VEGF) is the most potent angiogenic factor identified to date. TGF beta-1 acts as an indirect angiogenic agent. METHODS: VEGF and TGF beta-1 were measured in the serum of breast cancer patients and age-matched controls and in tumor tissue of cancer patients by ELISA. VEGF protein and mRNA expression by breast tumor cell lines were examined, and the effect of TGF beta-1 on VEGF production in these cells was assessed. RESULTS: VEGF levels were significantly higher (P = .03) in the serum of patients with breast cancer compared to age-matched controls. A positive correlation was found between serum (r = 0.539) and tumor tissue (r = 0.688) levels of VEGF and TGF beta-1. Metastatic MDA-MB-231 breast cancer cells produce more VEGF than do the primary BT474 cells. TGF beta-1 significantly (P < .05) increased production of VEGF. CONCLUSIONS: Breast cancer cells constitutively produce VEGF protein and mRNA. There is a relationship between VEGF and TGF beta-1 levels in breast cancer patients, and TGF beta-1 regulates VEGF expression by breast cancer cells. PMID- 9416409 TI - Phase III randomized trial of surgery with or without intraoperative photodynamic therapy and postoperative immunochemotherapy for malignant pleural mesothelioma. AB - BACKGROUND: Patients with malignant pleural mesothelioma (MPM) usually die of progressive local disease. This report describes the results of a Phase III trial comparing maximum debulking surgery and postoperative cisplatin, interferon alpha 2b, and tamoxifen (CIT) immunochemotherapy with and without intraoperative photodynamic therapy (PDT) to determine (1) whether such a multimodal approach can be performed with minimum morbidity and mortality in malignant pleural mesothelioma (MPM), and (2) whether first-generation (i.e., 630-nm laser light, Photofrin II) intrapleural PDT impacts on local recurrence of survival. METHODS: From July 1993 to June 1996, 63 patients with localized MPM were randomized to either PDT or no PDT. The tumors of 15 patients could not be debulked to 5 mm. Patients assigned to PDT (n = 25) and no PDT (n = 23) were similar with respect to age, sex, tumor volume, and histology. RESULTS: The type of resection (11 pleurectomies and 14 pneumonectomies vs. 12 pleurectomies and 11 pneumonectomies), length postoperative stay, and ICU time were comparable (PDT vs. no PDT). There was one operative death (hemorrhage), and each group had two bronchopleural fistulas. Postoperative staging divided patients into the following categories: stage I: PDT, 2, no PDT, 2; stage II: PDT, 2, no PDT, 2; stage III, PDT, 21; no PDT, 17; stage IV, PDT, 0; no PDT, 2. Comparable numbers of CIT cycles were delivered. Median survival for the 15 non-debulked patients was 7.2 months, compared to 14 months for the 48 patients on protocol. There were no differences in median survival (14.4 vs. 14.1 months) or median progression free time (8.5 vs. 7.7 months), and sites of first recurrence were similar. CONCLUSIONS: Aggressive multimodal therapy can be delivered for patients with higher stage MPM. First-generation PDT does not prolong survival or increase local control for MPM. PMID- 9416410 TI - Magnetic resonance cholangiopancreatography: a novel approach to the evaluation of suspected pancreaticobiliary neoplasms. AB - BACKGROUND: Magnetic resonance cholangiopancreatography (MRCP) is a new noninvasive diagnostic method for pancreaticobiliary (PB) imaging without endoscopy, sedation, or iodinated contrast. The purpose of this study was to evaluate the ability of MRCP to depict pancreatic and biliary ductal anatomy compared to that of endoscopic retrograde cholangiopancreatography (ERCP) and to evaluate the ability of MRCP to accurately diagnose PB neoplasms. METHODS: Twenty patients had MRCP, and 17 also had ERCP. All studies were read prospectively by experienced reviewers blinded to other imaging data. Pathologic diagnosis was made in all patients. RESULTS: Bile duct dilatation seen by ERCP in 14 of 17 patients was correctly identified by MRCP in all 14 patients, and normal ducts were correctly identified by MRCP in the other 3 patients. The pancreatic duct was visible on MRCP in the pancreatic head in 17 of 20 patients, the body in 17 of 20 patients, and the tail in 15 of 20 patients. At ERCP, pancreatic duct dilatation was present in 11 cases and was identified by MRCP in 10 of them. Eighteen of 20 patients had malignant PB neoplasms. MRCP indicated PB neoplasm in 19 patients. Seventeen of these 19 patients had histologically confirmed malignant neoplasms pathologically, whereas 2 had benign pathology (both chronic pancreatitis). Among the 17 patients who also had ERCP, MRCP and ERCP correctly agreed on a final diagnosis of malignant neoplasm in 14 cases. In the three cases in which MRCP and ERCP disagreed on a final diagnosis, MRCP was correct in one and incorrect in two. CONCLUSIONS: MRCP can accurately and noninvasively delineate PB ductal anatomy and diagnose PB neoplasms comparably to ERCP. MRCP is an interesting new noninvasive method for evaluating patients with suspected PB neoplasms. PMID- 9416411 TI - Differentiation of benign from malignant pancreatic masses by endoscopic ultrasound. AB - BACKGROUND: It is often difficult to determine whether a mass in the pancreas is benign or malignant. The goal was to evaluate whether endoscopic ultrasound (EUS) can reliably establish whether a mass is benign or malignant. METHODS: One hundred five patients with possible pancreatic tumors were referred for EUS. Those who were found to have a lesion suspicious for carcinoma and did not have a known malignancy also underwent EUS-guided FNA. RESULTS: A mass suspicious for cancer was identified in 73 patients, whereas inflammatory changes or a normal pancreas was noted in 32 patients. Four of the latter 32 patients were subsequently found to have cancer. EUS-guided FNA was performed on 47 of the 73 patients with a suspicious mass and was read as cancer in 27 patients, atypia in 10 patients, and benign in 10 patients. All 10 patients with atypia were subsequently confirmed to have cancer, and 6 of the 10 patients with a benign FNA were proved to have a tumor at surgery. EUS could differentiate the lesion as malignant with a sensitivity of 95%, specificity 88%, positive predictive value 95%, and negative predictive value 88%. CONCLUSIONS: Radial array EUS is helpful in supporting or refuting a diagnosis of cancer in a patient with a pancreatic mass. Although EUS-guided FNA can confirm the diagnosis, a negative FNA should not preclude exploration when clinically indicated. PMID- 9416412 TI - Cytologic evaluation of lumpectomy margins in patients with ductal carcinoma in situ: clinical outcome. AB - BACKGROUND: Breast conservation therapy is controversial for ductal carcinoma in situ (DCIS) due to recently reported high recurrence rates. We believe that cytologic evaluation of lumpectomy margins improves efficiency and leads to a lower recurrence rate following lumpectomy for DCIS. METHODS: A prospectively accrued database of 1255 breast cancer patients at the H. Lee Moffitt Cancer Center and Research Institute was found to have 218 patients with DCIS (17.4%). Of those 218 cases, 114 were treated with lumpectomy, axillary dissection, and radiation therapy; the remaining 104 patients were treated with mastectomy with or without reconstruction. Imprint cytology was used to evaluate all lumpectomy margins. Permanent sections and imprint cytology were reviewed by the same pathologist. RESULTS: All lumpectomy specimens (116 tumors in 114 patients) were evaluated. The median follow up was 57.5 months (range 2-110 months). One hundred and three patients with 104 tumors were selected on the basis of pure DCIS (with or without microinvasion), and treated with lumpectomy, axillary dissection and radiation therapy. Of the 104 tumors utilizing attempted breast conservation therapy, 7 (6.6%) required mastectomy. There were 6 recurrences (6.1%) with a median time for recurrence of 47.5 months (range 27-85 months); four recurrences were comedo and two were noncomedo at original diagnosis. CONCLUSIONS: The determination of lumpectomy margins in DCIS patients using imprint cytology leads to an overall recurrence rate of 6.1% with reduction in operative time, and re excision rate. Significant recurrence rates were associated with microinvasion and multifocal tumors (28%) versus simple DCIS at 5 years. Breast conservation therapy and surgical margin determination with imprint cytology for DCIS is a cost-effective and reliable method of treatment for simple DCIS. PMID- 9416413 TI - Role of conservation therapy for invasive lobular carcinoma of the breast. AB - BACKGROUND: Invasive lobular carcinoma (ILC) accounts for 5% to 10% of all invasive breast cancers. Although breast conservation therapy using local excision and postoperative irradiation is a standard therapy for early invasive ductal breast cancer, the result of this strategy in ILC is not well documented. We sought to determine the rate of locoregional recurrence after breast conservation therapy in patients with ILC. METHODS: A retrospective review of 74 patients with ILC treated with breast conservation therapy at The University of Texas M. D. Anderson Cancer Center (n = 43) or The John Wayne Cancer Institute (n = 31) between 1977 and 1993 was performed. RESULTS: The median age of patients was 60 years, and median follow-up was 56 months (range 1 to 207 months). Thirty nine patients had American Joint Committee on Cancer stage I disease, 30 had stage IIa disease, and five had stage IIb disease. All patients underwent surgical resection and postoperative radiation therapy. Twelve patients received postoperative adjuvant chemotherapy, and 27 patients were treated with adjuvant hormonal therapy. The 5-year actuarial locoregional recurrence rate was 9.8%, and the median time to recurrence was 77 months (range 41 to 113 months). Patients with positive or close (< or = 1 mm) surgical margins were at increased risk for local recurrence on univariate analysis (p = 0.034). Of the nine patients with breast recurrence, six underwent salvage therapy with total mastectomy and are disease free at the time of this writing, two patients died of distant disease, and one is alive with local disease at the time of this report. The 5-year disease-specific survival rate was 93.7%. CONCLUSIONS: Breast conservation therapy for ILC achieves locoregional control in the majority of patients. However, long-term follow-up of patients is important because many local recurrences following breast conservation therapy are late events. PMID- 9416414 TI - Age-related differences in breast cancer stage at diagnosis between black and white patients in an urban community hospital. AB - BACKGROUND: Breast cancer mortality is significantly higher among black patients compared to white patients. Black women are reportedly at increased risk for early-onset breast cancer. Our goal was to evaluate stage distribution relative to age among black and white breast cancer patients in an institution with a relatively high minority patient population. METHODS: We evaluated 425 patients diagnosed with breast cancer between 1990 and 1994: 56% white, 34% black, the remainder were other ethnicities. Patients were stratified by age: under 50 years versus 50 and older. Socioeconomic status was estimated by utilization of medical care in the private-practice setting versus the public clinic. RESULTS: Significantly more black patients were younger at diagnosis compared to white patients (32% vs. 20%; p = 0.008). There was a significantly more advanced stage distribution among the younger black patients, but not among the older black patients. Most of the black and white patients received private-practice care. CONCLUSIONS: These age-related differences in breast cancer stage distribution between black and white patients (which appeared independent of socioeconomic status) indicate that more aggressive screening and public education programs directed toward younger black women is warranted, and they lend support to the possibility of ethnicity-related variation in primary tumor biology. PMID- 9416415 TI - Retrospective review of 400 consecutive free flap reconstructions for oncologic surgical defects. AB - BACKGROUND: Free tissue transfer has become an important method for reconstructing complex oncologic surgical defects, allowing single stage reconstruction in most instances. The purpose of this study is to review a single center's experience with free flap reconstruction and describe trends that have evolved with respect to technique and donor site selection. METHODS: A retrospective review of 400 consecutive free flap reconstructions performed in 396 patients over 10 years was done. Regional applications include the head and neck (63%), trunk and breast (16%), lower extremity (16%), and upper extremity (5%). Donor sites include the fibula (109), rectus abdominis (93), forearm (72), latissimus dorsi (51), scapula (26), gluteus (25), jejunum (16), and five others (8). Microvascular anastomoses were performed to large-caliber vessels using a continuous suture technique; end-to-end anastomoses were preferred. Flaps were designed to avoid the need for vein grafts. Postoperative flap monitoring included clinical observation, conventional Doppler ultrasonography, surface temperature probes, and pinprick testing. RESULTS: The overall free flap success rate was 97%. Twenty-eight flaps (7%) were reexplored, of which seventeen were salvaged (61%). Surviving flaps resulted in a healed wound that did not delay postoperative radiation or chemotherapy. The complication rate was 14%. The mean duration of hospitalization was 21 days, with an average cost of $40,000. CONCLUSIONS: The use of fewer, reliable donor sites to reconstruct the vast majority of oncologic defects and the simplification of the microsurgical process have contributed to the success of free tissue transfer in this series. PMID- 9416416 TI - Inhibition of transforming growth factor alpha stimulation of human squamous cell carcinoma of the head and neck with anti-TGF-alpha antibodies and tyrphostin. AB - BACKGROUND: Transforming growth factor alpha (TGF-alpha) and its receptor (EGF-R) may regulate normal and malignant epithelial cell growth by an autocrine mechanism. We investigated the role of TGF-alpha in regulating head and neck SCC tumor growth. METHODS: TGF-alpha and EGF-R levels were measured in 7 SCC cell lines and 14 SCC biopsies by RIA, Scatchard, and Western analysis. TGF-alpha autocrine stimulation of DNA synthesis in SCC cell lines was assessed by incubation with TGF-alpha neutralizing antibodies and tyrphostin AG 1478, a selective and potent inhibitor of EGF-R kinase. RESULTS: All SCC cell lines synthesized TGF-alpha and expressed elevated EGF-R levels compared to normal keratinocytes. Twelve of the 14 SCC biopsies contained TGF-alpha protein and 8 had specific EGF-R. Exogenous TGF-alpha or EGF significantly increased DNA synthesis in 4 of 5 SCC cell lines. TGF-alpha neutralizing antibodies or tyrphostin AG 1478 reduced DNA synthesis in the two SCC cell lines (FaDu and SCC9) tested. CONCLUSIONS: These results indicate that SCC cell lines and tumors usually synthesize TGF-alpha, have elevated levels of EGF-R, and are mitogenically stimulated by a TGF-alpha autocrine system. Selective inhibition of the TGF-alpha system by EGF-R kinase inhibitors or TGF-alpha neutralizing antibodies may be useful strategies for treating SCC that overexpress TGF-alpha and its receptor. PMID- 9416417 TI - Chemical engineering of RNase resistant and catalytically active hammerhead ribozymes. PMID- 9416418 TI - Palladium-catalyzed synthesis of [E]-6-(2-acylvinyl)uracils and [E]-6-(2 acylvinyl)-1-[(2-hydroxyethoxy)methyl]uracils--their antiviral and cytotoxic activities. AB - [E]-6-(2-Acylvinyl)uracils and their corresponding 1-(2-hydroxyethoxy)methyl derivatives were synthesized through palladium-catalyzed reactions which involved an interesting rearrangement. Some of the acylvinyl uracils (3, 4, and 5) and the acyclonucleosides (8 and 10) showed pronounced activity against human T lymphocyte Molt 4/C8 and CEM cells. However, they were less toxic to murine L1210 and FM3A cells. The compounds did not have any marked antiviral activity. PMID- 9416419 TI - Alkylation of a catalytic aspartate group of the SIV protease by an epoxide inhibitor. AB - Specific irreversible inhibition of the SIV protease by FMOC-protected piperidine epoxide 1 involves alkylation of the protein. Tryptic digestion of the alkylated protein and mass spectrometric analysis of the peptides identify an active site aspartic acid (Asp-25) as the single residue that is alkylated. Computer modeling of 1 bound in the crystal structure of the SIV protease using DOCK 3.5 indicates that 1 has appropriate access to the active site. It is able to align in an orientation that allows a proton to be transferred to the epoxide from one of the catalytic aspartic acid groups in conjunction with nucleophilic attack on the epoxide of the carboxylate moiety of the second catalytic aspartic acid residue. Hydrophobic interactions are not optimal for this process due, in part, to the rigidity of the inhibitor ring system and the planar conformation of the amide. The combination of modeling with protein alkylation can provide insights into structural modifications of the inhibitor that may lead to improved inhibitory activity. PMID- 9416420 TI - TRH mimetics: differentiation of antiamnesic potency from antidepressant effect. AB - For the purpose of rational modification of the TRH molecule, we were pursuing an approach that consists of two steps: (1) 'obligatory' replacement of histidine with glutamine in TRH and (2) the application of conformational constraints for putative bioactive conformation I stabilized by an intramolecular hydrogen bond between C-terminal carboxamide proton and alpha-carbonyl of histidyl (glutaminyl), and conformation II formed by an intramolecular hydrogen bond between alpha-carbonyl of pyroglutamyl and prolinamide proton. Significant antiamnesic potency was discovered in the passive avoidance test (ECS and Scopolamine induced amnesia) for conformation II mimic (8S,10aS)-8-carbamoyl 1,2,3,6,7,8,9,10a- octahydro-5H,10H-pyrrolo[1,2-a][1,4]diazocin-5,10-dione (2) at doses of 0.1 and 1.0 mg/kg. EEG analysis indicates a mild activating effect of compound 2 on EEG, which is similar to that of piracetam and differs from hard amphetamine activation. Conformation I mimic 3-(2-carbamoylethyl)-2,3,6,7,8,8a hexahydro-1H,4H-pyrrolo[1,2-a] pyrazin-1,4-dione (1) exhibited an antidepressant effect at a dose of 1 mg/kg. The transition from two putative quasi-cyclic bioactive conformations of TRH and its obligatory similar analogue [Gln2]-TRH to their cyclic mimics led to differentiation of antiamnesic and antidepressant activity of TRH. PMID- 9416421 TI - Mechanism of degradation of 2'-deoxycytidine by formamide: implications for chemical DNA sequencing procedures. AB - We describe the reaction of formamide with 2'-deoxycytidine to give pyrimidine ring opening by nucleophilic addition on the electrophilic C(6) and C(4) positions. This information is confirmed by the analysis of the products of formamide attack on 2'-deoxycytidine, 5-methyl-2'-deoxycytidine, and 5-bromo-2' deoxycytidine, residues when the latter are incorporated into oligonucleotides by DNA polymerase-driven polymerization and solid-phase phosphoramidite procedure. The increased sensitivity of 5-bromo-2'-deoxycytidine relative to that of 2' deoxycytidine is pivotal for the improvement of the one-lane chemical DNA sequencing procedure based on the base-selective reaction of formamide with DNA. In many DNA sequencing cases it will in fact be possible to incorporate this base analogue into the DNA to be sequenced, thus providing a complete discrimination between its UV absorption signal and that of the thymidine residues. The wide spectrum of different sensitivities to formamide displayed by the 2' deoxycytidine analogues solves, in the DNA single-lane chemical sequencing procedure, the possible source of errors due to low discrimination between C and T residues. PMID- 9416422 TI - The influence of molecular conformation upon the self-assembly of cyclohexane diamide diacids. AB - BACKGROUND: Information regarding the self-association of small peptide motifs can be used in the design of peptide microstructures. Previous work in our laboratories illustrated the self-association of certain diamide diacids into microcapsules. In this report a series of cyclohexane diamide diacids are investigated. The cyclohexylene (R-C6H10-R) system (with its axial and equatorial requirements) provided an opportunity to study the influence of molecular conformation upon the self-aggregation process. RESULTS: Condensation of the respective cis- and trans-1,2-, 1,3-, and 1,4- cyclohexane dicarboxylic acid platforms with two equivalents of a L-Phe ester followed by deprotection gave the desired diamide diacids. Basic solutions of cis-1,2-, trans-1,3-, and cis-1,4 diamide diacids generated solid microspheres when acidified to pH 2.4. Molecular modeling revealed that 1,3-diaxial interactions favor a helical turn within these diamides. CONCLUSIONS: Access to 'complementary' molecular geometries is needed to self-associate into microscopic architectures. PMID- 9416423 TI - Conformational preference for segetalins G and H, cyclic peptides with estrogen like activity from seeds of Vaccaria segetalis. AB - Three-dimensional structures in DMSO-d0 of segetalins G [cyclo(-Gly-Val-Lys-Tyr Ala-)] and H [cyclo(-Gly-Tyr-Arg-Phe-Ser-)], cyclic pentapeptides from seeds of Vaccaria segetalis, showing estrogen-like activity, were determined by the distance geometry calculation and restrained energy minimization from NMR data. The backbone structure of segetalin G contains one beta-turn: a beta II-like turn at Tyr4-Ala5, and that of segetalin H one beta-turn: a beta II' turn at Gly1-Tyr2 and one gamma-turn at Arg3-Phe4-Ser5 sequence. The results of distance comparison analysis proposed a pharmacophore model of estrogen-like cyclic peptides, segetalins. PMID- 9416424 TI - Synthesis and antibacterial activity of novel 4-pyrrolidinylthio carbapenems--I. 2-Alkoxymethyl derivatives. AB - The synthesis and in vitro antibacterial activity of a novel series of 2 alkoxymethyl-4-pyrrolidinylthio-1 beta-methyl carbapenems are described. As a result of these studies, we discovered that FR27743 (19j) containing a novel 2 fluoroethoxymethyl substituent possesses a broad spectrum of antibacterial activity against both Gram-positive and Gram-negative organisms, including Pseudomonas aeruginosa. Furthermore, FR27743 exhibited excellent stability against renal dehydropeptidase-I (DHP-I), good urinary recovery, and superior in vivo activity compared to that for Meropenem against several systemic infections. PMID- 9416425 TI - A novel class of potent gamma-aminobutyric acid aminotransferase inhibitor, 3 (hydroxyamino)propylamine and analogues. AB - Hydroxyamino analogues of gamma-aminobutyric acid (GABA) were synthesized and evaluated for inhibitory activity toward gamma-aminobutyric acid aminotransferase (GABA-T). The title compound, 3-(hydroxyamino)propylamine (HPA), showed a potent inhibitory activity. The inhibition is competitive with respect to GABA and the Ki value of GABA-T for HPA is 0.4 mmol. The activity of inhibition is comparable to those of aminoxyacetic acid and valproic acid. 3 (Hydroxyaminomethyl)piperidine (3HMP), a cyclic analogue of HPA, also showed a potent inhibitory activity, whereas 3-(methoxyamino)propylamine (OMe-HPA), 3-(N hydroxy-N-methylamino)propylamine (NMe-HPA) and 4-(hydroxyamino)piperidine (4HP) showed weak activity. PMID- 9416426 TI - Tumor necrosis factor-alpha production-inhibiting activity of phthalimide analogues on human leukemia THP-1 cells and a structure-activity relationship study. AB - N-Substituted phthalimides (2-substituted 1H-isoindole-1,3-diones) were prepared and their inhibitory effects on tumor necrosis factor-alpha (TNF-alpha) production by human leukemia cell line THP-1 stimulated with 12-O tetradecanoylphorbol 13-acetate (TPA) or okadaic acid (OA) were examined. A structure-activity relationship study of these phthalimide analogues revealed that their inhibitory effects on TPA- and OA-induced TNF-alpha production by THP 1 cells are well correlated to each other, i.e. they may involve the same target molecule(s). An analysis by the use of phthalimide analogue-immobilized affinity gels indicated the existence of several phthalimide-binding proteins in THP-1 cell extract. PMID- 9416427 TI - Inhibition of papain with 2-benzyl-3,4-epoxybutanoic acid esters. Mechanistic and stereochemical probe for cysteine protease catalysis. AB - Papain, a prototypic cysteine protease was inactivated by methyl and benzyl esters of (2S,3S)-2-benzyl-3,4-epoxybutanoic acid. On the other hand, methyl ester of (2S,3R)-2-benzyl-3,4-epoxybutanoic acid was shown to be a competitive inhibitor for the enzyme. It was inferred from the inactivation stereochemistry that in the papain catalytic reaction the nucleophilic attack of the side chain thioalkoxide of Cys-25 on the scissile peptide bond of substrates occurs in the 're' fashion. The papain inactivating potency of (2S,3S)-2-benzyl-3,4 epoxybutanoic acid methyl ester was enhanced over three-fold in a pH 8.0 solution compared with in the neutral solution. This together with our previous observation with alpha-chymotrypsin and the recent theoretical treatment of the enzymic reaction of papain, suggest that in the inactivation of papain by oxirane containing inhibitors, the oxirane does not need to be activated by prior protonation as thought previously. The oxirane ring is sufficiently labile that the unprotonated oxirane moiety can undergo an electrophilic reaction with the Cys-25 thiolate. PMID- 9416428 TI - Surfactant protein B deficiency: insights into inherited disorders of lung cell metabolism. PMID- 9416429 TI - T-cell regulation in murine and human autoimmune diabetes: the role of TH1 and TH2 cells. AB - Insulin-dependent diabetes mellitus (IDDM) results from the destruction of pancreatic insulin-secreting cells by a T-cell-mediated autoimmune reaction. Distinct types of T helper cells (TH1 and TH2) have been characterised based on their cytokine secretion profiles following activation. Evidence from animal models favours the hypothesis that autoimmune diabetes is a TH1 response. However, there is no clear indication that a primary imbalance between protective TH2 and deleterious TH1 cells at early stages can trigger the autoimmune process. Protective CD4 + cells detected in nondiabetic young non-obese diabetic mice have not been shown to work through TH2 cytokines. In humans, there is little evidence that IDDM results from a TH1 response. Indeed, efficient experimental systems are lacking in humans to study the regulation of the autoimmune response in vitro. Interestingly, several immunotherapy strategies have aimed at inducing a TH2 response, even though TH2 cells have not been implicated in spontaneous disease development. However, recent ongoing trials in humans using oral administration of insulin to prevent diabetes are based on a protective mechanism which seems to depend essentially on transforming growth factor-beta. This cytokine is not dependent on TH1/TH2 dichotomy. Thus, although several attempts have been made to induce a TH1/TH2 switch to obtain a protective effect, a different and more complex mechanism probably (and paradoxically) accounts for the oral protection actually tested in animal models and humans. PMID- 9416430 TI - Different TH2-TH1 balance in V beta 8 and V beta 6 subsets of splenocytes in NOD females in the early phase of diabetogenesis. AB - Non-obese diabetic (NOD) mice spontaneously develop T-cell-mediated autoimmune diabetes. Initial work on the diabetogenic T-cell repertoire indicated that autoreactive T lymphocytes were polyclonal but that the presence of specific subsets (V beta 8 or V beta 6) might be required for induction of the disease. Further functional analysis of NOD mice T lymphocytes was limited because of the relative anergic state of these cells due to abnormal patterns of cytokine secretion. The purpose of the present study was to establish experimental conditions allowing the exploration of the functional features of minor T lymphocyte subsets in vitro using low doses of cofactors. The ability of splenocytes to proliferate, respond to, or secrete interleukin-2 and interleukin 4 was explored in young, pre-diabetic or old non-diabetic female NOD mice. No significant bias in T-cell receptor usage was noted in the spleen of these animals, whereas V beta 6 + lymphocytes could be very efficiently stimulated by interleukin-4 and also produce low but detectable amounts of interleukin-4 during the pre-diabetic period in female NOD mice. These results suggest that diabetes induction is preceded by V beta + subset-specific functional changes in the ability of various T cells to respond to or secrete interleukin-2 and interleukin 4, indicating a functional imbalance of the T-cell repertoire expanded by the autoimmune process. PMID- 9416431 TI - A five-year study of the incidence of insulin-dependent diabetes mellitus in young Tunisians (preliminary results). AB - Three Tunisian districts were selected to estimate the incidence of insulin dependent diabetes mellitus (IDDM): Beja, Monastir and Gafsa. A population-based registry for new cases of IDDM was established in 1990 in these three areas according to WHO DIAMOND project methodology. A local extension of the protocol consisted in the inclusion of children up to 19 years of age. Children with a diagnosis of IDDM discharged from general hospitals and private clinics in these areas were recorded in the corresponding registry. A secondary source of case ascertainment was provided by regional school health centers. The findings of the five-year study showed that 156 cases of IDDM were recorded among children aged 0 to 19 years in the three regions. The degree of ascertainment was estimated at 96%. The global age-adjusted incidence rates were 6.76.100,000(-1) year-1 and 6.95.100,000(-1).year-1 in the 0 to 14- and 0 to 19-year age-groups respectively. Age-adjusted incidence rates were lower in Monastir than in Beja and Gafsa, respectively 4.69, 8.13 and 8.33.100,000(-1).year-1 for subjects aged 0 to 19 years. Incidence rates showed no significant difference by gender but were lower in the 0 to 4- and higher in the 10 to 14-year age groups. No time trend was detected. Sixty-two percent of all cases were diagnosed in the cold season. The incidence rate of IDDM in Tunisia is thus close to that observed in most Mediterranean countries. PMID- 9416432 TI - Insulin sensitivity and beta-cell function in essential hypertension and normotensive first-degree relatives of hypertensive subjects. AB - We investigated glucose metabolism and beta-cell function in normotensive subjects with one essential-hypertensive parent and in subjects with mild/moderate essential hypertension (eHT) before and after 12-week treatment with nitrendipine. The hypertensive-parent group comprised 12 normotensive subjects, and the hypertensive group 15 subjects with mild/moderate eHT. A corresponding control group composed of 20 normotensive subjects was also investigated. All subjects underwent a frequently sampled intravenous glucose tolerance test (FSIGT). Hypertensive subjects underwent FSIGT testing before and after 12 weeks of treatment with nitrendipine (20 mg per day). Insulin sensitivity, glucose effectiveness and beta-cell function were investigated using the minimal model technique on the basis of FSIGT test data. No significant differences were detected in any of the minimal-model parameters either between the hypertensive-parent group and the control, or between the hypertensive group (before nitrendipine treatment) and the control. Twelve weeks of anti hypertensive treatment with nitrendipine led to an increase in glucose effectiveness and a non-significant increase in glucose tolerance, but had no significant effects on other minimal-model parameters or on the serum lipid profile. Our results suggest that eHT cannot be considered consistently associated with insulin resistance. Nitrendipine treatment appears to have no undesirable effects on peripheral sensitivity to insulin or on beta-cell function. However, the 12-week course led to a 72% increase in glucose effectiveness. PMID- 9416433 TI - Silent myocardial ischaemia and left ventricle hypertrophy in diabetic patients. AB - The purpose of this study was to evaluate the ability of three noninvasive techniques to detect silent myocardial ischaemia and analyse the factors associated with this condition, particularly left ventricular hypertrophy, in diabetic patients. An ECG stress test, a thallium-201 myocardial scintigraphy with dipyridamole intravenous infusion, ambulatory 48 h ECG monitoring and an echocardiographic study were performed in 92 diabetic patients without cardiac symptoms but with > or = 2 additional cardiovascular risk factors. At least one of these tests was positive in 28 patients (30.4%), suggesting silent myocardial ischaemia. Twenty-four of these patients had a coronary angiography which showed significant coronary stenosis in only 9 cases. An accurate echocardiographic tracing was obtained in 79 patients, particularly in 7 of the 9 with coronary stenosis. Left ventricular hypertrophy was detected in 34 patients, 6 of whom had coronary stenosis. In patients with left ventricular hypertrophy, the positive predictive values of myocardial scintigraphy and the ECG stress test were respectively 50% and 100%, as compared to only 33% and 11% in those without hypertrophy. In summary, coronary stenoses were found in < 10% of asymptomatic diabetic patients with > or = 2 cardiovascular risk factors, but more frequently in individuals with left ventricular hypertrophy. Thus, silent myocardial ischaemia should be searched for first in diabetic patients with hypertrophy, for which the stress test was the most accurate detection method in this study. PMID- 9416434 TI - Insulin-like growth factor binding proteins from adult-hamster pancreatic islets: influence of glucose concentration. AB - This study investigated the effect of glucose on insulin-like growth factor binding proteins (IGFBPs) in islets isolated from pancreas of adult hamsters and compared the response pattern with that of their serum IGFBPs. Serum samples and islets were obtained from adult normal male hamsters, and IGF-binding capacity was measured in aliquots of serum, sonicated islets, or conditioned medium using either 125I-hIGF-I or -II. IGFBPs were characterized in these samples by the ligand-blotting technique, and insulin was measured in conditioned medium by radioimmunoassay. Three IGFBP fractions were identified in serum, with relative molecular weights of 38, 30-33, and 24 kDa, while only two fractions of 30-33 and 24 kDa were identified in islets or in their conditioned medium. Islets cultured with 2 or 16 mM glucose for 48 h released more insulin in the presence of the higher glucose concentration. The binding capacity measured in the islet suspension or conditioned medium increased as a function of glucose concentration in the incubation medium. The IGFBPs present both in islets and conditioned medium had a 3- to 4-fold higher apparent affinity for IGF-II than IGF-I. The higher glucose concentration increased the intensity of the two IGFBP bands identified in the islet suspension by 2- to 3-fold. Our data show that two low molecular-weight IGFBPs were released from adult hamster pancreatic islets, with a different distribution pattern from that of hamster serum, and that the amount of IGFBPs released by islets depended on the glucose concentration in the culture medium. Though not conclusive, these data suggest that IGFBPs may play a regulatory role in B-cell turnover in adult islets as they do in foetal islets. PMID- 9416435 TI - Gender effect of the Trp64Arg mutation in the beta 3 adrenergic receptor gene on weight gain in morbid obesity. AB - Phenotypic expression of the Trp64Arg mutation in the beta 3-adrenoceptor gene (beta 3-AR) has been found to be somewhat variable among different populations, suggesting that it may be influenced by genetic background and environmental factors. As sex may also influence gene allellic expression, we evaluated a potential gender effect of the Trp64Arg mutation in 292 morbidly obese subjects [body mass index (BMI) > or = m/kg2]. Although the 15 mutated obese females were younger than the non-mutated ones, the difference between their current weight and their weight at 20 years was significantly higher (62.4 +/- 20.0 kg versus 47.0 +/- 24.0 kg; p = 0.017). Moreover, in the mutated heterozygous female group, the mean Zscore (individual BMI minus reference French population mean BMI/SD of reference population BMI) was significantly higher (8.0 +/- 2.5 versus 6.0 +/- 2.0 SD of BMI, p = 0.0018), as was the maximal Zscore calculated from the maximal BMI that obese females reached during life (9.0 +/- 3.0 versus 7.0 +/- 2.5, p = 0.005). The regression curves of the Zscore against age showed that the curve of mutated females was shifted to the top, indicating that their BMI was higher regardless of age. These effects were not observed in the male group (the Zscore was 6.7 +/- 3.0 vs. 7.2, p = 0.7 respectively in mutated and non-mutated men). These data reinforce the hypothesis that the expression of the beta 3-AR susceptibility gene depends on additional factors including gender and possibly hormonal status. PMID- 9416436 TI - Proposed criteria for the diagnosis of diabetes: evidence from a French epidemiological study (D.E.S.I.R.). AB - The American Diabetes Association and a working group of the International Diabetes Federation and the World Health Organisation are considering the possibility of lowering the level of fasting glucose for diagnosis of diabetes from 7.8 mM to 7.0 mM (126 mg/dl) and adding a "hyperglycaemia" category (6.1-6.9 mM). This report studied the resulting change in the frequency of diabetes in the French D.E.S.I.R. cohort and evaluated cardiovascular risk factors according to the proposed limits. The frequency of treated diabetes was 1.0% in this cohort of more than 5,000 French men and women 30 to 64 years of age, 0.7% had a fasting glucose > or = 7.8 mM versus 1.6% for the proposed > or = 7.0 mM. Using the new criteria, 3.8% of men and 1.6% of women would be diabetic. For many cardiovascular risk factors, men with fasting glucose > or = 7.8 mM had a significantly higher risk than those in the (7.0-7.7 mM) range, whereas no significant differences were found for women. There were few differences between the (7.0-7.7 mM) and (6.1-6.9 mM) ranges, but highly significant differences were apparent for both sexes between the normoglycaemic (< 6.1 mM) and hyperglycaemic (6.1-6.9 mM) categories. The percentage of men with two or more of ten risk factors increased with fasting glucose: 36% in the normoglycaemic (< 6.1 mM) and 49% in the hyperglycaemic (6.1-6.9 mM) categories; 73% with fasting glucose in (7.0-7.7 mM) versus 90% with fasting glucose > or = 7.8 mM. In women, the corresponding percentages were lower: 26% and 60%, 32% versus 64% respectively. The major increase in cardiovascular risk factors would appear to occur at 6.1 mM, for men, whereas the increase in the combination of risk factors was at the more conservative cut-off of 7.0 mM. PMID- 9416437 TI - Measurement of substrate oxidation in man. AB - The measurement of substrate oxidation in living individuals can facilitate metabolic investigations. Indirect calorimetry and tracer techniques allow such measurements. Indirect calorimetry provides simultaneous calculation of the rate of oxidation of the three major macronutrients (carbohydrates, fat and protein) from respiratory gas exchanges and urinary nitrogen excretion. Such estimates represent net substrate oxidation rates. Thus, carbohydrate oxidation represents the oxidation of either endogenous glycogen or exogenously administered carbohydrate. It also includes de novo lipogenesis (with simultaneous oxidation of lipids in amounts equivalent to their synthesis), but does not include oxidation of glucose formed from gluconeogenesis from amino acids or glycerol. The accuracy of these calculations depends on the adequacy of the stoichiometry used for oxidation of substrates, which has to be varied when special forms of substrate are used. Tracer techniques consist in administration in tracer amounts of a selected nutrient labelled with 14C or 13C and monitoring of the specific activity/isotopic enrichment of the substrate in plasma and of the pulmonary elimination of labelled CO2. Such techniques allow assessment of the actual rate of oxidation of one substrate at a time. However, there are major pitfalls relating to the recovery of labelled CO2 in breath during oxidation of the substrate as well as during non-steady state conditions. PMID- 9416438 TI - Functional outcome following thalamic haemorrhage: relationship between motor and cognitive functions and ADL. AB - Twenty-two patients with thalamic haemorrhage were examined to investigate the relationship between motor and cognitive function, and activities of daily living (ADL). Patients with unilateral spatial neglect had lower ADL scores on admission than patients without unilateral spatial neglect (Mean: 17.0 and 24.6, respectively; F = 4.38, df = 1, p < 0.05). Unilateral spatial neglect related to feeding, bowel control and transfer in Barthel index on admission. Patients with aphasia on admission had lower ADL at discharge than patients without aphasia on admission (Mean: 57.0 and 84.7, respectively; F = 7.70, df = 1, p < 0.05). Aphasia related to the bathing, toilet, stair climbing, dressing, and ambulation in Barthel index on discharge. There was a significant difference between the severity of paresis in upper and lower limb on admission and ADL at discharge. The two-way repeated measures ANOVA showed a significant difference between severity of paresis in lower limb and ADL improvement. It can be suggested that the most important predictor of outcome was paresis in lower limb, and not aphasia or unilateral spatial neglect. PMID- 9416439 TI - The Edmans ADL index: validity and reliability. AB - The Edmans ADL index was developed to assess functional abilities in stroke patients, including the activities necessary to enable a person to live independently at home, and graded to enable staff to monitor patient's progress over time. Content validity was established by comparing the Edmans ADL index with other published ADL assessments. Construct validity was established by comparing the Edmans and Barthel ADL indices, for 60 patients admitted consecutively to the Nottingham stroke unit. This showed a strong association between assessments. Inter-rater reliability was assessed by two occupational therapists, who independently and simultaneously assessed 20 patients individually on the stroke unit. This showed 96% excellent/good agreement between observers. Test-retest reliability was established by assessing 20 patients, 1 year post-stroke, on two separate occasions, 1 month apart. Results showed 85% excellent/good agreement over time. We conclude that the Edmans ADL index has content and construct validity, is sensitive to change over time, and has inter rater and test-retest reliability. PMID- 9416440 TI - Models of disability: a critical perspective. PMID- 9416441 TI - Disability models in geriatrics: comprehensive rather than competing models should be promoted. AB - The usefulness of different models of disability is discussed. There is no clear cut demarcation between ability and disability, and a person's functional abilities are highly dependent on societal as well as individual factors. One should not, however, promote models of disability that cover only the social aspects, but rather try to build comprehensive models including medical, psychological and social aspects of disability. The World Health Organization's International Classification of Impairments, Disabilities and Handicaps (ICIDH) provides a useful basis for such model building. The main weakness of the ICIDH is that it fails to take the subjective perceptions of the individual fully into account. Accordingly, it should be supplemented by some model of subjective well being. Possible relationships between subjective well-being and the ICIDH concepts are discussed. PMID- 9416442 TI - Models of disability for children. PMID- 9416443 TI - Models of disability. Definitions as power. PMID- 9416444 TI - Models of disability: an example from ophthalmology. PMID- 9416445 TI - Models of disability. Who needs models? PMID- 9416446 TI - Afghan children and mental retardation: information, advocacy and prospects. AB - The family and community situation of children with mental retardation (learning difficulties/disabilities) is discussed within the Afghan cultural heritage and the current realities of civil war and refugee villages. Some formal services are being developed for children with physical and visual impairments. Mental retardation and hearing impairments are comparatively neglected, as skills are lacking and progress tends to be much slower. Some casual integration occurs in ordinary schools and kindergartens, which should be supported. Experiences in family counselling can be developed and also extended by enlisting indigenous counsellors and healers. The major strategy for the foreseeable future will be support and enhancement of family resources. PMID- 9416447 TI - From laboratory to clinic: a large scale study of distortion product otoacoustic emissions in ears with normal hearing and ears with hearing loss. AB - OBJECTIVES: 1) To describe distortion product otoacoustic emission (DPOAE) measurements in large groups of subjects with normal hearing and with hearing loss, and to use these data to provide comprehensive descriptions of DPOAE test performance. 2) To describe the effects of primary frequency and audiometric threshold on the extent to which DPOAE measurements accurately identify auditory status. 3) To develop an approach that describes the probability that any measured response is coming from either a normal or an impaired ear. 4) To develop an approach for representing DPOAE data clinically. 5) To explore the relation between magnitude of hearing loss and DPOAE measurements. DESIGN: DPOAE measurements were made in 1267 ears of 806 subjects, using stimulus conditions that previously had been demonstrated to result in the greatest separation between normal and impaired ears (i.e., primary levels of 65/55 dB SPL for f1/f2; Stover et al., 1996). Subjects were recruited from local clinical populations and through local advertisements. All data were analyzed using clinical decision theory, including relative operating characteristic (ROC) curves and estimates of areas under these curves (Az). In addition, cumulative distributions were constructed of response properties from both normal and hearing-impaired ears. These cumulative distributions were used to select specific probabilities that measured responses were coming from either the normal or impaired distributions, and to develop an approach for describing clinical DPOAE data. RESULTS: For no conditions were the distributions of DPOAE responses from normal and impaired ears completely separated, meaning that optimal criterion values would still result in errors in identification of auditory status. Test performance, defined by Az, was best for mid and high frequencies and poorest for lower frequencies and for the highest frequency tested (8000 Hz). Performance was best when normal hearing was defined as audiometric thresholds between 20 and 30 dB HL, with poorer performance for more stringent or lax audiometric criteria. CONCLUSIONS: Within the limits related to the effects of primary frequency and audiometric criterion, it appears that DPOAE measurements can be used to accurately identify auditory status. An approach is described, using the present data set, that allows one to assign to any measured DPOAE value (DPOAE amplitudes, DPOAE/noise) the probability that the response is coming either from the distribution of normal or impaired responses. In addition, DPOAE/noise systematically decreases as hearing loss increases over the range of hearing losses from 0 to about 40 to 60 dB HL (depending on frequency), thus potentially enabling one to differentiate hearing losses over this range. For hearing losses greater than 50 to 60 dB HL, ears do not produce measurable DPOAEs and thus, no predictive relationship exists. PMID- 9416448 TI - Speech perception performance of nucleus multichannel cochlear implant users with partial electrode insertions. AB - OBJECTIVE: The present investigation examined the speech perception performance of five children with ossified cochleas who received partial insertions of the Nucleus 22-channel cochlear implant. DESIGN: The partial-insertion subjects' preimplant and 1.5 yr postimplant performance on a battery of speech perception tests was compared to the average performance of age-matched control subjects who received full electrode insertions. All the partial-insertion subjects were fit with their Nucleus cochlear implant between the ages of 2 and 5 yr, and had used their device for at least 1.5 yr. More extended comparisons also were made for the two partial-insertion subjects who had used their cochlear implants for a longer period of time. RESULTS: The subjects with partial electrode insertions performed similarly to the control group at both the preimplant and 1.5 yr postimplant intervals. Furthermore, the partial-insertion subjects showed continued improvements in speech perception performance with increased device experience past 1.5 yr, again similar to the full-insertion control group. CONCLUSIONS: The present results suggest that partial insertion of a multichannel implant device is an appropriate and feasible approach to the surgical management and auditory rehabilitation of children with extensive or complete ossification of the cochlea. PMID- 9416449 TI - The predictive value of measures of preverbal communicative behaviors in young deaf children with cochlear implants. AB - OBJECTIVE: To determine whether measures of early communicative behavior in young children obtained within the first year after implantation could predict speech and language skills measured 3 yr after implantation. DESIGN: An unselected sample of 17 children receiving multichannel cochlear implants under the age of 5 yr were monitored over a 3 yr period. During the first year after implantation, the development of early communicative behavior was measured at intervals using an established video analysis technique. Three years after implantation, four outcome measures were taken: the IOWA closed-set sentence test, a measure of continuous discourse tracking ability, a rating of ability to use the telephone and a global rating of auditory performance. RESULTS: There was a significant correlation between the video analysis measure taken at the 12 mo interval, specifically the extent of auditory and vocal behaviors, and three of the outcomes obtained at the 3 yr interval: both performance-based measures and the telephone rating. When the four outcomes were aggregated to form a composite measure, the correlation was highly significant. There was no significant correlation between the composite measure and the video analysis measures obtained earlier than the 12 mo interval. CONCLUSION: Development of a predominantly auditory and vocal style of early communicative behavior is predictive of relatively high levels of skill on speech and language tasks measured 2 yr later. PMID- 9416450 TI - Effects of formant bandwidth on the identification of synthetic vowels by cochlear implant recipients. AB - OBJECTIVE: The main objective was to investigate whether the broadening and narrowing of formant bandwidths had a significant effect on the identification of vowels often confused by Nucleus cochlear implant recipients using the Spectral Peak (SPEAK) speech coding strategy. Specifically, identification performance for synthetic vowels with the first two formants (F1 and F2) parametrically varied in bandwidth was explored. DESIGN: Eight implanted subjects identified synthetic versions of the isolated vowel sounds [I, epsilon, lambda, [symbol: see text]] with F1 and F2 bandwidth manipulations, as well as foil tokens of [i, u, a, ae, [symbol: see text]]. Identification performance was examined in terms of percent correct as well as error patterns. Further analyses compared patterns of electrode activation. RESULTS: In general, broader F1 bandwidths yielded poorer performance and narrower F1 bandwidths yielded better performance relative to identifications for the reference stimuli. However, similar manipulations of F2 bandwidths resulted in less predictable performance. Comparison of electrode activation patterns indicated a distinct sharpening or flattening in the F1 frequency region for subjects with the greatest performance extremes. CONCLUSIONS: Manipulation of F1 bandwidth can result in concomitant changes in electrode activation patterns and identification performance. This suggests that modifications in the SPEAK coding strategy for the F1 region may be a consideration. Similar manipulations of F2 bandwidth yielded less predictable results and require further investigation. PMID- 9416451 TI - Changes in synthetic and natural vowel perception after specific training for congenitally deafened patients using a multichannel cochlear implant. AB - OBJECTIVE: The aim was to determine whether the ability to use place-coded vowel formant information could be improved after training in a group of congenitally deafened patients, who showed limited speech perception ability after cochlear implant use ranging from 1 yr 8 mo to 6 yr 11 mo. A further aim was to investigate the relationship between electrode position difference limens and vowel recognition. DESIGN: Three children, one adolescent, and one young adult were assessed with synthesized versions of the words/hid, head, had, hud, hod, hood/containing three formants and with a natural version of these words as well as with a 12-alternative, closed-set task containing monosyllabic words. The change in performance during a nontraining period was compared to the change in performance after 10 training sessions. RESULTS: After training, two children showed significant gains on a number of tests and improvements were consistent with their electrode discrimination ability. Difference limens ranged from one to three electrodes for these patients as well as for two other patients who showed minimal to no improvements. The minimal gains shown by the final patient could be partly explained by poorer apical electrode position difference limen. CONCLUSIONS: Significant gains in vowel perception occurred post-training on several assessments for two of the children. This suggests the need for children to continue to have aural rehabilitation for a substantial period after implantation. Minimal improvements, however, occurred for the remaining patients. With the exception of one patient, their poorer performance was not associated with poorer electrode discrimination. PMID- 9416452 TI - Auditory stimulus intensity and reaction time in listeners with longstanding sensorineural hearing loss. AB - OBJECTIVE: This study examined relationships among sound level, subjective loudness, and reaction time in listeners with longstanding sensorineural hearing loss and loudness recruitment. DESIGN: A simple reaction-time test was performed by 10 hearing-impaired (HI) subjects with varying degrees of recruitment and by 10 normal-hearing (NH) control subjects. Both groups listened to 0.5 kHz tones presented at a soft level, representing the soft endpoint of a subject's functional dynamic range, and a loud level representing the loud endpoint. In one condition, the loudness level of stimuli within a block was fixed, and hence predictable; in another it varied randomly within a block between loud and soft, adding uncertainty to the simple reaction time task. A test with exactly the same design but using visual stimuli was also performed. RESULTS: 1) In general, the HI subjects responded to auditory stimuli with normal or near-normal mean reaction times when stimulus loudness was predictable. 2) Introduction of uncertainty disrupted the reaction time performance of some, but not all, of the HI subjects. 3) Both the HI and the NH subjects responded more quickly to loud tones than to soft ones. Under the predictable loudness condition the magnitude of this "speed up" was nearly identical for the two groups despite the fact that the HI group had, on average, only half the physical dynamic range of the NH group. 4) Visual reaction-time performance was equivalent between the HI and NH groups in all important respects. CONCLUSIONS: Despite long-term auditory deficits, HI subjects' ability to respond quickly to simple auditory signals is not substantially impaired, particularly when listening under predictable loudness conditions. Although physical soft-loud ranges will generally be narrower than normal for HI subjects, their reaction time performance at the endpoints of that range is likely to be near normal. PMID- 9416453 TI - Childhood hearing impairment: processing dependencies in multidimensional speech perception for an auditory level of analysis. AB - OBJECTIVE: Our purpose was to determine whether the influence of childhood hearing impairment (HI) on multidimensional speech processing is a purely linguistic effect or whether childhood HI also affects the processing of speech dimensions representing an auditory level of analysis. DESIGN: The processing dependencies characterizing the two dimensions of talker-gender and spatial location were studied in 40 children with HI and in two normal-hearing (NH) comparison groups representing similar chronological ages (N = 30) or similar vocabulary skills (N = 70). The processing interactions were assessed with a speeded selective-attention task requiring listeners to attend selectively to the gender of the talker and to ignore the spatial location and vice versa. The logic is that performance for the target dimension will not be affected by what is happening on the nontarget dimension if the dimensions are processed independently. On the other hand, if the dimensions are not processed independently, listeners will not be able to attend selectively and performance for the relevant dimension will be affected by what is happening on the irrelevant dimension. In the latter case, results may be analyzed in terms of Garner interference (the effect on performance of irrelevant variability in the to-be-ignored dimension) (Garner, 1974a) and Simon interference (the effect on performance of an irrelevant spatial source) (Simon, 1990). RESULTS: Overall results in all listeners, those with NH or HI, showed significant interference when the participants were attending to the gender of the talker and ignoring spatial location and vice versa. The talker-gender and spatial-location dimensions of speech were not processed independently by these children. When the processing interactions were compared between the NH and HI groups, the presence of childhood HI as a general rule significantly diminished the degree of interference from spatial location. The degree of interference from the gender of the talker, on the other hand, remained normal in the presence of childhood HI. All listeners seemed stimulus bound by the gender of the talker. The degree of Garner interference did not show age-related or degree of loss-related change. The degree of Simon interference showed significant change as a function of age in the children with mild-moderate HI, but not in the children with severe HI. The developmental function for Simon interference in the children with mild moderate HI was delayed to a degree that corresponded to the duration of the auditory deprivation. CONCLUSIONS: The overall pattern of results indicates that the influence of childhood HI on multidimensional speech processing is not a purely linguistic effect. PMID- 9416454 TI - Vigabatrin in refractory childhood epilepsy. The Brazilian Multicenter Study. AB - Children, 47, with various types of severe drug-resistant epilepsy were entered into a prospective, add-on, open trial with vigabatrin. Patients with West syndrome and idiopathic generalized epilepsies were excluded. Seven children had the drug withdrawn, five because of increase in seizure frequency and two because of adverse effects. Drug efficacy, measured according to seizure type, showed a 100% decrease in seizure frequency in 18.6% of partial seizures and 17.3% of the generalized seizures. There was a higher than 50% decrease in 39.5% of partial and 60.8% of generalized seizures, and less than 50% decrease or increase in seizure frequency in 41.8% and 21.8% of partial and generalized seizures, respectively. Vigabatrin mean dosage during phase 3 was 63.6 mg/kg per day (S.D. = 30.5), ranging from 19.3 to 110.5 mg/kg per day. Parametric statistical analysis (Student's t-test) of seizure frequency between phases 1 and 3 showed a significant decrease in seizure frequency for partial (P = 0.022), and generalized seizures (P < 0.0001). Drug-related adverse effects were observed in 18/47 cases (38.3%), consisting mainly of irritability, hyperactivity, dizziness, somnolence and gastrointestinal symptoms. PMID- 9416455 TI - Epilepsy with myoclonus and post-natal development of the motor system in humans: a hypothesis. AB - Epilepsy with myoclonus is thought to be linked to the motor system. At birth, development of the central nervous system in humans is far from being achieved. Post-natal changes take place at different levels in this neuronal system. These modifications suggest that the motor cortex is a highly dynamic structure during post-natal development. They may account for the age-dependence of various epileptic syndromes. (1) The number of synapses increases during the early post natal years and then decreases to reach the adult level around puberty. (2) Neurons differentiate and synthesize various neurotransmitters. (3) Dendrites grow actively and participate in the formation of local cortical circuits. (4) Electrophysiological properties of cortical neurons change during the first months of rodent development. This could reflect modifications of the ion channels present in the cell membrane. (5) The pyramidal tract myelinate and exuberant collaterals are selectively removed. These two processes are dependent on neuronal electrical activity. It has been demonstrated that selective collateral stabilization is promoted by glutamate release and stimulation of the N-methyl-D-aspartate (NMDA) receptor. So, seizures occurring during the neonatal period may interact with these normal developmental features. Furthermore, neuronal electrical activity and seizures stimulate the transcription of specific messenger RNAs coding for neurotrophic factors like nerve growth factor (NGF) or brain-derived neurotrophic factor (BDNF). The overproduction of neurotrophic factors leads to maldevelopment of the cortex. PMID- 9416456 TI - Interictal and ictal activity in the rat cobalt/pilocarpine model of epilepsy decreased by local perfusion of diazepam. AB - We investigated the efficacy of focal perfusion of diazepam (DZP) in reducing seizures produced by focal cobalt and systemic pilocarpine in the rat. Cobalt chloride crystals (3.5 mg/kg) were inserted stereotactically into the left hippocampus and recording electrodes affixed to the head of 23 rats. Focal spiking was evident within 5-7 days of implantation. Occasional ictal electrographic events were observed with cobalt alone, but consistent ictal events could be produced by intraperitoneal injection of pilocarpine hydrochloride (60 mg/kg) into the cobalt-treated animals. When rhythmical spiking was observed, the animals were treated either with DZP (0.25 mg in 50 microliters) or a vehicle (VEH) delivered into the left hippocampus. Blinded spike counts before and after injection showed spiking at 133.3 +/- 53.4% of baseline (mean +/- SD, n = 8) for the VEH-treated animals and 2.7 +/- 3.3% (n = 8) for the DZP-treated animals. Ictal events occurred in seven of the eight VEH treated and two of the eight DZP-treated rats. Mean time to the first ictal event was 5.9 +/- 6.9 min for VEH-treated animals and 24 +/- 32.6 min for DZP-treated animals. DZP injected into the hippocampus contralateral to the cobalt did not reduce spiking. Systemic levels of DZP were unmeasurable in nine of ten tested animals. Focal perfusion of DZP therefore effectively reduced spiking in this cobalt chloride/pilocarpine model of focal and secondarily generalized epilepsy. This model, while involving GABAergic mechanisms, does not entirely depend upon GABAergic mechanisms. The findings therefore broaden the possibility of using focal DZP as a treatment for partial seizures. PMID- 9416457 TI - Effects of a subconvulsant dose of kainic acid on afterdischarges elicited by cortical stimulation in rats. AB - The aim of this study was to test the hypothesis that nonconvulsive seizures elicited by a low dose of kainic acid may induce acute as well as chronic changes in brain function. Cortical epileptic afterdischarges (ADs) characterized by spike-and-wave rhythm and clonic seizures of facial and forelimb muscles were elicited in adult male rats with chronically implanted electrodes. Four stimulations were given in each of four weekly sessions. In the second session, 26 animals were injected with kainic acid (6 mg/kg i.p.) and 19 rats received no injection. The acute effects of kainic acid were to increase the intensity of movements accompanying stimulation and abruptly prolong ADs. Epileptic ADs were followed by a depression of electrocorticographic activity in both noninjected and kainic acid groups. In addition, when kainate was administered, interictal spike activity was registered mostly in the occipital region. One and two weeks after kainate administration, i.e. in the third and fourth stimulation sessions, there was an increased incidence of transitions from spike-and-wave ADs to another, limbic type of afterdischarge. This functional change persisted although no obvious neuronal death was found in the hippocampi of 12 other rats that received the same dose of kainic acid. PMID- 9416458 TI - The influence of established and new antiepileptic drugs on visual perception. 1. A placebo-controlled, double-blind, single-dose study in healthy volunteers. AB - The influence of single oral dosages of carbamazepine (CBZ), valproic acid, vigabatrin (VGB), lamotrigine (LTG), gabapentin (GBP), and losigamone (LSG) on visual perception was investigated in ten healthy volunteers according to a double-blind, placebo-controlled, cross-over study design. The test battery comprised visual acuity, the Lanthony-D-15-desature colour perception test, increment, postadaptation and transient tritanopia threshold measurements, perception threshold assessment for monochromatic and chromatic gaussian dots, monochromatic gratings and gratings of differing spatial frequency, and critical flicker fusion tests with various stimuli. The only consistent and partly significant effects were seen after VGB and GBP. After VGB, increment, postadaptation and transient tritanopia thresholds and the critical flicker fusion increased, whereas GBP led to a somewhat converse profile. The other tests were not influenced consistently by any antiepileptic drug (AED). We conclude that: (i) gamma-amino-butyric acid-(GABA)-related properties as under the prototype drug VGB result in specific alterations of the transient tritanopia phenomenon which is consistent with the physiological hypothesis for this retinal paradigm based on extracellular recordings in primates. The possible mechanisms why VGB improved critical flicker fusion as the only AED in this trial are discussed. The profile of GBP indicates a unique mechanism of action. We have not observed specific influences on visual perception under AEDs which act mainly via alterations of ion membrane conductance. The transient tritanopia and flicker fusion paradigms we used appear to be promising to investigate antiepileptic drugs with hitherto unknown modes of actions in human noninvasively. PMID- 9416459 TI - The influence of established and new antiepileptic drugs on visual perception. II. A controlled study in patients with epilepsy under long-term antiepileptic medication. AB - In this study, we investigated visual performance under chronic antiepileptic drug treatment. Patients were under carbamazepine (CBZ) (n = 18), valproic acid (VPA) (n = 9), CBZ and vigabatrin (VGB) (n = 4), CBZ and gabapentin (GBP) (n = 8), and under CBZ and topiramate (TPR) (n = 6), respectively. Seven untreated patients with epilepsy and 42 healthy volunteers served as controls. The test battery comprised the Lanthony-D15-desature colour perception test, increment, postadaptation and transient tritanopia (TT) threshold measurements, visual perception threshold assessments for monochromatic and chromatic gratings and gaussian dots, and critical flicker fusion (CFF) tests. No differences were seen between naive patients and healthy controls. Patients under drug treatment always showed alterations of visual perception. Postadaptation and TT thresholds were altered under each drug regimen after short delays between switching off the adaptation light and switching on the blue test light. Threshold elevations were maximum under the combination of CBZ and TPR and lowest under CBZ and GBP. Consistent impairment of the CFF was seen under combined CBZ and TPR whereas VPA as well as combined CBZ and VGB led to ameliorations the mechanisms of which are discussed. The other tests were less sensitive. In conclusion, alterations of visual function were apparent under chronic antiepileptic drug treatment both with established and new agents. However, it may be difficult to distinguish between effects based on specific modes of action and nonspecific retino- and neurotoxicity. PMID- 9416460 TI - Effects of AWD 140-190 on stimulus-induced field potentials and on different patterns of epileptiform activity induced by low calcium or low magnesium in rat entorhinal cortex hippocampal slices. AB - AWD 140-190 a potent new anticonvulsant was tested on several types of epileptiform activities in entorhinal cortex hippocampal slices. AWD 140-190 suppressed completely and in a dose-dependent manner spontaneous seizure-like events induced by lowering extracellular Ca2+. In the low magnesium model, AWD 140-190 applied with 200 microM reduced recurrent short discharges in area CA1 by 48.1 +/- 14.7%, while in the entorhinal cortex seizure-like events were not depressed. Late recurrent discharges were increased in frequency to 213.8 +/- 78.1 and reduced in amplitude by 50.1 +/- 14.4%. Responses to paired pulse stimuli with intervals ranging from 20 to 150 ms were reduced both with alvear and stratum radiatum stimulation. Decreases in [Ca2+]0 and associated slow field potentials evoked by repetitive stimulation of stratum radiatum were also depressed in a dose-dependent manner. AWD 140-190 also reduced stimulus-induced rises in [K+]0. AWD 140-190 200 microM diminished the amplitude of slow field potentials observed during high K(+)-induced spreading depression by about 17% in CA1 and 34% in entorhinal cortex without any significant effect on SD-associated rises in [K+]0. These results suggest that AWD 140-190 has an anticonvulsant effect presumably by interfering with repetitive generation of action potentials. AWD 140-190 may also possess modulatory effects on glial cells as suggested by the strong depression of SD-associated slow negative potential shifts. PMID- 9416461 TI - Kainic acid-induced generalized seizures alter the regional hippocampal expression of the rat m1 and m3 muscarinic acetylcholine receptor genes. AB - We investigated the gene expression responses using in situ hybridization with radiolabelled riboprobes for the m1 and m3 subtypes of muscarinic cholinergic receptors in the rat hippocampus following a brief (5-min) kainic acid-induced behavioral seizure. The kainic acid was intraperitoneally administered, and the ensuing generalized convulsive seizure terminated with diazepam. Our results demonstrate that the expression of the m1 subtype was significantly reduced in the CA1, CA3 and the dentate granule cells by 3 h after the administration of kainic acid while no significant change was observed in any hippocampal subfield for the m3 subtype. By 6 h post challenge, the m1 subtype was still decreased in all hippocampal subfields examined, while the m3 subtype remained unchanged from vehicle injected control. At 24 h post challenge, both the m1 and m3 subtypes were significantly reduced in the CA1 and CA3 subfields; the expression of the m1 subtype in the dentate granule cells, however, had recovered to levels indistinguishable from vehicle-injected control. These results demonstrate that epileptiform activity induced by kainic acid administration promotes alterations in the expression levels for both the m1 and m3 muscarinic receptor genes, and suggest that the activity of this neuromodulatory system in the hippocampus may be altered through activity-dependent mechanisms at early times following seizures. PMID- 9416462 TI - Muon spectroscopy applied to biological systems: a study of thiyl radicals, RS. AB - Thiyl radicals (RS.) are formed when positive muons (mu+) are implanted into solutions of thiocarbonyl compounds (C = S) in ethanol, tetrahydrofuran or formamide solvents. A solvent dependence is found, which reflects changing electronic interactions and overall conformations, suggesting that the properties of RS. radicals may be dependent on the polarity (philicity) of the environment in which they are formed. PMID- 9416463 TI - Inhibition of NADPH oxidase-mediated superoxide radical formation in PMA stimulated human neutrophils by 4-hydroxynonenal--binding to -SH and -NH2 groups. AB - 4-Hydroxynonenal (HNE), a major lipid peroxidation product, effectively inhibits the superoxide radical formation by NADPH oxidase of phorbol myristate acetate (PMA)--stimulated human PMNL. The I50 value for the inhibition of NADPH oxidase mediated superoxide radical formation by 4-hydroxynonenal was found to be 19 microM. The HNE inhibition involves the reaction with both -SH and -NH2 groups. Superoxide formation as final result of the NADPH oxidase cascade was almost completely restored by addition of dithiothreitol. In presence of hydroxylamine only a minor restoration of superoxide radical formation was found. A combination of dithiothreitol and hydroxylamine yielded the greatest recovery. Two other aldehydes with the same chain length as HNE but different binding to lysine, histidine and cysteine residues, trans-2,3-nonenal and nonanal, gave I50 values for the inhibition of NADPH oxidase-mediated superoxide formation rate of 110 microM or > 300 microM, respectively. PMID- 9416464 TI - Leukocyte mobilization, chemiluminescence response, and antioxidative capacity of the blood in intestinal ischemia and reperfusion. AB - Intestinal ischemia and reperfusion elicits changes in leukocyte counts and increased production of reactive oxygen species (ROS). The purpose of this study was to investigate whether these changes were followed by and/or connected with changes in the extracellular antioxidative capacity in a rat superior mesenteric artery (SMA) occlusion/reperfusion model. The SMA was occluded for 45 min and then allowed to be reperfused. Changes of leukocyte, polymorphonuclear (PMN), and lymphocyte counts, chemiluminescence (CL) of whole blood samples as a marker of ROS production, and the total antioxidative capacity of the serum were quantified at the end of ischemia and in 1 h intervals during the postischemic period up to 4 h. The myeloperoxidase (MPO) activity in the serum and intestinal tissue samples was also determined. The MPO activity in the intestinal tissue samples was significantly elevated at the end of ischemia, and this elevation lasted for the whole postischemic period. The oxidative challenge to the body induced a fast mobilization of extracellular antioxidative mechanisms already at the end of ischemia, which was followed by a significant increase in PMN counts and whole blood CL starting at the 2nd hour after reperfusion. The increased CL activity of whole blood was attributed to the increase of the circulating PMNs. No significant changes were observed in leukocyte and lymphocyte counts. It is concluded that compensatory mechanisms of the oxidative-antioxidative balance of the body react very quickly if challenged. PMID- 9416465 TI - Decreased antioxidant defense mechanisms in rat liver after methanol intoxication. AB - The primary metabolic fate of methanol is oxidation to formaldehyde and then to formate by enzymes of the liver. Cytochrome P-450 and a role for the hydroxyl radical have been implicated in this process. The aim of the paper was to study the liver antioxidant defense system in methanol intoxication, in doses of 1.5, 3.0 and 6.0 g/kg b.w., after methanol administration to rats. In liver homogenates, the activities of Cu,Zn-superoxide dismutase and catalase were significantly increased after 6 h following methanol ingestion in doses of 3.0 and 6.0 g/kg b.w. and persisted up to 2-5 days, accompanied by significant decrease of glutathione reductase and glutathione peroxidase activities. The content of GSH was significantly decreased during 6 hours to 5 days. The liver ascorbate level was significantly diminished, too, while MDA levels were considerably increased after 1.5, 3.0 and 6.0 g/kg b.w. methanol intoxication. Changes due to methanol ingestion may indicate impaired antioxidant defense mechanisms in the liver tissue. PMID- 9416466 TI - Reaction of substituted anthracenes and a butadiene with nitric oxide: product formation determined by EPR spectroscopy. AB - When nitric oxide (NO), generated from nitric acid and copper, was reacted with a series of 9,10-substituted anthracenes, only 9,10-dimethylanthracene gave EPR detectable nitroxide radicals, although the expected bicyclic nitroxide arising from cheletropic NO addition across the 9,10-positions was not observed. Purity of the NO is crucial as the presence of higher oxides of nitrogen leads to radicals by hydrogen abstraction which are trapped by NO and the resultant nitroso compounds produce stable nitroxides detectable by EPR. In contrast the acyclic system, 3,4-diphenyl-2,5-dimethyl-2,4-hexadiene gives rise to an EPR spectrum consistent with cheletropic NO addition, although higher oxides of nitrogen again mediate the formation of different nitroxides. PMID- 9416467 TI - Protective effect of ACE inhibitors on ischemia-reperfusion-induced arrhythmias in rats: is this effect related to the free radical scavenging action of these drugs? AB - The antiarrhythmic effects of captopril, a sulphydryl-containing angiotensin converting enzyme (ACE) inhibitor, were compared with those of the nonsulphydryl containing ACE inhibitor lisinopril and the sulphydryl-containing agent glutathione in an in vivo rat model of coronary artery ligation. To produce arrhythmia, the left main coronary artery was occluded for 7 min, followed by 7 min of reperfusion. Captopril (3 mg kg-1) and lisinopril (0.1, 0.3 or 1 mg kg-1) caused marked decreases in mean arterial blood pressure (BP) and heart rate, whereas glutathione (5 mg kg-1) had no effect on them. The incidence of ventricular tachycardia (VT) on ischemia and reperfusion was significantly reduced by captopril and lisinopril. Captopril and 1 mg kg-1 lisinopril also significantly decreased the number of VEB during occlusion and the duration of VT on reperfusion, respectively. These drugs also attenuated the incidence of reversible ventricular fibrillation (VF) and the number of ventricular ectopic beats (VEB) during reperfusion. However, glutathione only reduced the incidence of VT on reperfusion, significantly. These results suggest that, in this experimental model, ACE inhibitors limit the arrhythmias following ischemia reperfusion and free radical scavenging action of these drugs does not have a major contributory role in their protective effect. PMID- 9416468 TI - Separation and characterization of cholesteryl oxo- and hydroxy-linoleate isolated from human atherosclerotic plaque. AB - In previous work we demonstrated that up to 30% of cholesteryl linoleate in homogenates of advanced human plaque samples is present in oxidized forms. Here we show that the material from plaque hexane extracts which co-elutes with cholesteryl hydroxy-linoleate on reversed phase HPLC (Anal Biochem 1993;213:79), is composed of several isomers of cholesteryl hydroxy- and cholesteryl oxo octadecadienoate. Enzymatic hydrolysis and measurement of liberated cholesterol and disappearance of the esters revealed that almost all of the material consisted of unoxidized cholesterol esterified to oxidized derivatives of octadecadienoate. Semi-preparative reversed-phase HPLC was used to obtain sufficient quantities of this co-eluting material to undertake normal phase HPLC separation of these components. The nature of such separated and isolated compounds was identified, by co-chromatography with authentic standards, UV spectroscopy and chemical ionization and electron impact mass spectrometry, as cholesteryl hydroxy- and cholesteryl oxo-octadecadienoate. These oxidized fatty acids have been observed previously in plaque, in agreement with our new unambiguous demonstration of their presence as cholesteryl esters. The application of the methods described for the separation of the various forms of oxidized cholesteryl octadecadienoate may aid mechanistic studies of in vitro and in vivo lipoprotein lipid oxidation. PMID- 9416469 TI - DNA single strand breaks by aromatic nitroso compounds in the presence of thiols. AB - Aromatic nitroso compounds, nitrosobenzene (NB), N,N-dimethyl-4-nitrosoaniline (DMNA) and 3,5-dibromo-4-nitrosobenzene sulfonate (DBNBS), caused DNA single strand breaks in the presence of thiol compounds. The strand breaking was inhibited completely by free radical scavenger ethanol. Electron spin resonance (ESR) studies showed that hydronitroxyl (or sulfur-substituted nitroxyl) radicals were generated in the early stage of the interactions. Formation of these radicals was not inhibited by ethanol, indicating that these radicals did not directly contribute to the strand breaking. The DNA strand breaking was inhibited partially by superoxide dismutase and catalase under the limited conditions, but not by removal of oxygen from or addition of metal chelators to the reaction mixture. By ESR-spin trapping technique using 5,5-dimethyl-1-pyrroline-N-oxide (DMPO), the DMPO-OH spin adduct was detected. Formation of the spin adduct was inhibited by superoxide dismutase and catalase. The hydronitroxyl (or the sulfur substituted nitroxyl) radicals may reduce oxygen into active oxygen species and also transformed by themselves into other unidentified free radical species to cause the DNA strand breaks. PMID- 9416470 TI - Hydroxyl radical scavenging activity of nonsteroidal anti-inflammatory drugs. AB - The hydroxyl radical (.OH)-scavenging activity of d-2-[4-(3-methyl-2 thienyl)phenyl]propionic acid (M-5011), a novel nonsteroidal anti-inflammatory drug (NSAID), and that of several other NSAIDs were investigated by the hyaluronic acid (HA) degradation method and the electron spin resonance (ESR) spin-trapping technique. The superoxide anion (.O2-)-scavenging activity of M 5011 was also measured by the ESR technique. (1) M-5011 and the other NSAIDs examined inhibited the degradation of HA induced by the Fenton reaction system in a dose-dependent manner. (2) M-5011 and the other NSAIDs scavenged .OH directly in a dose-dependent manner. (3) M-5011 was the most potent drug among the NSAIDs tested regarding the scavenging activity of .OH as follows; M-5011 > indomethacin > ketoprofen = suprofen > aspirin. The .OH-scavenging activity of M-5011 was potent in comparison with that of oxidized glutathione (GSSG), an endogenous .OH scavenger. (4) M-5011 did not scavenge .O2-; nor did GSSG. These results suggest that M-5011 acts as a scavenger of .OH at sites with inflammatory lesions. PMID- 9416472 TI - Medical devices and the year 2000 problem. AB - The "year 2000 (Y2K) problem"--the fact that microprocessor-based systems or software may malfunction when the year changes from 1999 to 2000--has received considerable media attention. Solutions to the problem have not always been swift in coming, however, and the healthcare industry is one area in which there is still much to do. In the May 1997 issue of Health Devices (26[5]), ECRI published a Talk to the Specialist article that briefly described the possible effects of the year 2000 date change on medical equipment. Since that time, we have gathered additional information and developed new recommendations and guidelines for dealing with this situation. PMID- 9416471 TI - Antioxidant properties of the major polyphenolic compounds in broccoli. AB - We have examined the antioxidant activity of the major phenolic compounds in Broccoli: two flavonol glycosides (quercetin 3-O-sophoroside and kaempferol 3-O sophoroside) and four hydroxycinnamic acid esters (1,2'-disinapoyl-2-feruloyl gentiobiose, 1-sinapoyl-2-feruloyl gentiobiose, 1,2,2'-trisinapoyl gentiobiose and 1,2-disinapoyl gentiobiose). The Trolox C equivalent antioxidant capacity (TEAC) and inhibition of iron/ascorbate-induced lipid peroxidation of phosphatidyl choline vesicles were measured. In the aqueous phase TEAC assay, the two flavonol glycosides were less active than their respective aglycones. TEAC values for the hydroxycinnamic acid esters were less than the sum of their constituent hydroxycinnamic acids on a molar basis. Quercetin 3-O-sophoroside was a potent inhibitor of lipid peroxidation, in contrast to kaempferol 3-O sophoroside. The hydroxycinnamic acid esters were highly effective at preventing lipid damage with the exception of 1,2,2'-trisinapoyl gentiobiose. The six compounds analysed herein demonstrate the antioxidant activity of the major phenolics in broccoli and indicate the effect on antioxidant activity of sugar substitutions in the phenolic B ring. PMID- 9416473 TI - GE HiSpeed advantage computed tomography scanner. AB - Computed tomography (CT) scanners use x-rays to produce thin cross-sectional images, or slices, of human anatomy. Their applications include diagnosing head and body masses, assessing bone mineral content, displaying vasculature, and detecting and evaluating stenosis in blood vessels. Currently, three different CT scanning methods are available: (1) conventional CT; (2) spiral scanning, in which information is collected in a single continuous scan rather than in discrete slices; and (3) ultrafast CT, in which a rotating x-ray beam is produced without the movement of mechanical components. ECRI conducted this Evaluation at a hospital that had just taken delivery of a new GE Medical Systems HiSpeed Advantage CT scanner, a third-generation unit that can perform conventional or spiral scanning. This study, which was performed in conjunction with acceptance testing, provides the essential information required to ensure adequate system performance. The capabilities that we tested include tomographic slice geometry, radiation output, image quality, and human factors design. Readers are cautioned not to base purchasing decisions on our rating of this device alone, but on a thorough understanding of the issues surrounding CT scanners and the factors used to develop our conclusions, which can be acquired only by reading this study in its entirety. Reviewing the entire study will also help readers to gain the perspectives necessary to judge units other than the one we have evaluated. PMID- 9416474 TI - Continued risk of entrapment between the siderails and canopy top in some older cribs. PMID- 9416475 TI - Mounting screw short-circuits bed-check alarm system. PMID- 9416476 TI - False apnea alarm on a Protocol Propaq physiologic monitor. PMID- 9416477 TI - Intermittent operation of Elekta Selector ultrasonic aspirators. PMID- 9416478 TI - Diagnosis of biliary ductopaenia. PMID- 9416479 TI - Retrieved endogenous biotin: a novel marker and a potential pitfall in diagnostic immunohistochemistry. AB - AIM: Antigen retrieval (AR) procedures are based on the effect of heating (by either microwave or pressure cooking treatments) on routinely fixed and paraffin embedded tissues. We observed that AR procedures restore the reactivity of endogenous biotin (EB) and report on the distribution of EB following AR in a series of routinely fixed and embedded tissues. METHODS AND RESULTS: Following pressure cooking or microwave treatments, a simple streptavidin-peroxidase staining revealed retrieved endogenous biotin (REB) in normal tissues (such as liver, kidney and adrenal cortex), in oxyphylic cells and in some tumours, especially in carcinomas of the kidney and of the adrenal cortex. In formalin fixed (but not in alcohol-fixed) tissue sections, the heating procedures caused an intense and finely granular cytoplasmic reaction, following a routine streptavidin-conjugated peroxidase treatment. The staining was prevented by blocking of EB by a sequential avidin-biotin treatment. CONCLUSIONS: Retrieval of EB reactivity can cause pitfalls in diagnostic immunohistochemistry but, alternatively, it might also constitute a useful and novel diagnostic marker. PMID- 9416480 TI - Identification of oncocytic lesions of salivary glands by anti-mitochondrial immunohistochemistry. AB - AIMS: To evaluate the immunohistochemistry using an anti-mitochondria antibody in the investigation of various oncocytic lesions of the salivary glands. METHODS AND RESULTS: Ten cases of adenolymphoma (Warthin's tumour) and one case each of benign oncocytoma and oncocytic carcinoma of the salivary glands were examined. Normal salivary glands were also tested. They were investigated immunohistochemically using mouse monoclonal antibody against human mitochondria. In normal salivary glands, epithelial cells of the striated ducts showed a thick linear immunoreactivity, which corresponded well to the intracytoplasmic distribution pattern of mitochondria. In addition, a small number of swollen epithelial cells showing an intense, finely granular immunoreactivity in the cytoplasm were scattered in the ductal system and acini ('oncocytic metaplasia'). Almost all neoplastic cells involved in adenolymphoma, benign oncocytoma, and oncocytic carcinoma showed an intense, finely granular immunoreactivity in the cytoplasm. CONCLUSIONS: Immunohistochemistry using the anti-mitochondria antibody proved to be a highly sensitive and specific method for light microscopic identification of mitochondria and superior to routine H & E or PTAH stain especially in the detection of isolated oncocytic cells. PMID- 9416481 TI - Immunohistochemical analysis of decalcified paraffin-embedded human bone marrow biopsies with emphasis on MHC class I and CD34 expression. AB - AIMS: To identify the stromal structures and haematopoietic cell lineages in normal bone marrow. The optimal conditions were studied for the reactivity of a panel of antibodies, applicable to paraffin sections of decalcified trephine biopsies using antigen retrieval methods. METHODS AND RESULTS: Two methods of antigen retrieval (pepsin and acid citrate buffer) were tested. For the demonstration of most antigens and for reduction of background staining, heating in acid citrate buffer was preferred. In the case of elastase and von Willebrand Factor (factor VIIIrAg) pepsin pre-treatment was optimal, whereas Ulex europaeus agglutinin (UEA-1) and alpha-smooth muscle actin (alpha-SMA) required no pretreatment. Staining patterns obtained after 48 h EDTA decalcification and short electrolytic decalcification were identical. Both methods allowed recognition of HLA-A and HLA-B antigens, isolated CD34+ cells, mono-histiocytic cells (CD68+), myeloid cells (elastase and myeloperoxidase), erythroid cells (glycophorin C) and of megakaryocytic cells (Factor VIIIrAg). A relative simple lymphocyte-subset analysis was possible in decalcified paraffin sections allowing recognition of B-cells (CD20+) and T-cells (CD3+ and CD45RO+) in frequencies comparable to frozen sections. Suitable stromal cell staining was achieved by vimentin and desmin antibodies, whereas the bone marrow capillary network was visualized by CD34, factor VIIIrAg and UEA-1. CONCLUSIONS: This immunohistochemical study indicates that all cellular components of the haematopoietic microenvironment of the bone marrow can be identified in decalcified paraffin sections using antigen retrieval methods and that the time of decalcification can be reduced to 1-1.5 h. PMID- 9416482 TI - Stereological estimates of the nuclear/cytoplasmic ratio and star volume on fibreoptic biopsies are of prognostic value for survival in a preliminary study of advanced squamous cell carcinoma of the lung. AB - AIMS: This study evaluated the role of morphometric and clinical parameters in establishing the prognosis of patients submitted to radiotherapy for advanced squamous cell carcinoma of the lung. METHODS AND RESULTS: Morphometric studies were performed by point counting techniques. Forty patients were included in this study. Group 1 patients (n = 22) were those with survival equal to or less than 6 months; group 2 (n = 10) patients had a survival of 7 to 12 months; and group 3 (n = 8) included patients with survival greater than 12 months. To characterize these three groups of patients, models combining categorical and continuous variables were constructed by means of discriminant analysis. Weight loss, histological grade, nuclear/cytoplasmic ratio and star volume of the nuclei were selected during the backward procedure as relevant variables to characterize the three groups of patients. The overall sensitivity of the model was 90%. CONCLUSIONS: Our results indicate that histopathological data may help to predict prognosis in patients with advanced squamous cell lung carcinoma, and encourage the use of morphometric procedures in histopathological analysis of this type of lung tumour. PMID- 9416483 TI - bcl-2 and p53 immunophenotypes in pre-invasive, early and advanced oesophageal squamous cancer. AB - AIMS: An inverse correlation between bcl-2 and p53 expression has been reported in several types of epithelial tumour. The role of bcl-2 and p53 in the development of oesophageal squamous carcinoma has yet to be established. The expression of bcl-2 and p53 proteins has been evaluated in the multistage oesophageal tumorigenesis, which progresses from normal mucosa to dysplasia (squamous intraepithelial lesion, SIL), to invasive early and advanced oesophageal squamous cancer. METHODS AND RESULTS: Sixty-four cases of squamous oesophageal cancer, coexisting with SIL in 18 cases, were immunohistochemically analysed for any overexpression of bcl-2 and p53 proteins. Any association of bcl 2 and p53 protein expression with patient survival was also analysed. We observed bcl-2 expression that decreased significantly during the progression of oesophageal carcinogenesis. A decreasing frequency in the expression of bcl-2 in advanced oesophageal squamous cancer coincided with frequent p53 overexpression. bcl-2 expression was correlated with patient survival by univariate analysis. The association disappeared after adjusting for tumour stage, p53 overexpression showed no association with patient survival by either univariate or multivariate analysis. CONCLUSIONS: The down-regulation of bcl-2 and upregulation of p53 in advanced oesophageal squamous cancer suggest that bcl-2 and p53 proteins may interact in the progression of oesophageal squamous cancer. PMID- 9416484 TI - Expression of c-Met correlates with grade of malignancy in human astrocytic tumours: an immunohistochemical study. AB - AIMS: Recent studies suggest the involvement of hepatocyte growth factor/scatter factor (HGF/SF) in glioma cell invasion and tumour progression. We investigated the distribution and rate of tumour cells that express c-Met protein, which is the cell-surface receptor for HGF/SF, in astrocytic tumours. The type of cells that express c-Met in tumour tissues was also identified. METHODS AND RESULTS: c Met expression was screened immunohistochemically in a total of 43 astrocytic tumours, including 14 low-grade astrocytomas (A), 13 anaplastic astrocytomas (AA) and 16 glioblastoma multiforme (GBM), c-Met reactivity was demonstrated predominantly in the cytoplasm of tumour cells. Bizarre large tumour cells tended to stain intensely. Higher c-Met expression levels (> or = 2+, more than 25% cells were positive) were noted in 21.4% of (A) vs. 53.8% in (AA) and 87.5% in (GBM) (P < 0.001), indicating a clear relationship between c-Met protein staining and higher grade astrocytic tumours. Moreover, c-Met immunoreactivity was also shown in tumour microvasculature, reactive astrocytes, and neurones in the cortex infiltrated by glioma cells. In 85.7% of cases containing infiltrated cortex, neurones were positive vs. no neurones in non-neoplastic regions (P < 0.002). CONCLUSIONS: This evidence suggests that c-Met expression in the brain could be associated with astrocytoma progression and also reactive process. Immunohistochemical determination of c-Met-expressing cell types helps to understand possible roles of c-Met in tumour tissues. PMID- 9416485 TI - Nasal T/NK cell lymphomas commonly express perforin and Fas ligand: important mediators of tissue damage. AB - AIMS: Two molecular mechanisms of T/natural killer (NK) cell-mediated cytotoxicity, one perforin based and the other Fas based, have been demonstrated, and both systems induce cytotoxicity in the target cells. The Fas-based mechanism involves the transducing molecule Fas and its ligand (FasL). In addition, perforin and/or FasL are also expressed in the cytotoxic T/NK cells. This study was thus designed to investigate the Fas and perforin pathways of the cytotoxic T/NK lymphoma cells in the nasal cavity. METHODS AND RESULTS: Eight patients with nasal lymphoma were analysed using immunohistochemical staining methods. Two cases were CD3+ CD56+ (T/NK cell) type, and six were CD3- CD56+ (NK cell) type. All cases showed Epstein-Barr virus genomes by in-situ hybridization. In addition, all cases showed the expression of TIA-1 (GMP-17), which is a marker of cytotoxic T and NK cells. FasL was expressed in the majority of the lymphoma cells and some histiocytes, while Fas was found in lymphoma cells and many non neoplastic cells. In addition, the expression of perforin was detected in almost all lymphoma cells. In the double stainings, lymphoma cells expressed both FasL and perforin. Based on these findings, both the perforin- and Fas-based pathway of the cytotoxic T/NK lymphoma cells are thus considered to play an important role in the clinical features. CONCLUSIONS: Tissue damage is a common morphological feature in nasal T/NK cell lymphoma. The above findings therefore support the theory that tissue damage is due to both the cytotoxicity of T/NK lymphoma cells as well as to angiocentricity. PMID- 9416486 TI - CD44 expression is up-regulated in the deep zone of osteoarthritic cartilage from human femoral heads. AB - AIMS: The objective of this study was to detail the topographical and zonal distribution of the cell adhesion molecule CD44 in normal and osteoarthritic cartilage. METHODS AND RESULTS: Immunohistochemistry utilizing well characterized anti-CD44 antibodies (clones A3D8, Bric 235, 2C5) was performed on cryostat and paraffin sections of human articular cartilage from macroscopically normal (n = 18) and osteoarthritic (n = 11) femoral heads. Samples for cryostat sections were obtained from 12 topographically different sites. Sections were divided into zones (superficial, middle, deep) and the CD44 staining scored. Chondrocytes in normal articular cartilage and cartilage from osteoarthritic femoral heads stained positive for CD44 in both cryostat and paraffin sections. Normal cartilage showed a significant decrease in CD44 staining in the deep zone as compared to the superficial zone (P < 0.05). However, cryostat sections of residual cartilage from osteoarthritic femoral heads showed increased CD44 staining in the deep zone as compared to normal articular cartilage. The CD44 staining showed no topographical variation in either the normal cartilage or the osteoarthritic residual cartilage. CONCLUSIONS: CD44 expression displays a distinct zonal variation in normal articular cartilage which is lost in osteoarthritic cartilage due to an up-regulated expression in the deep zone. CD44 expression does not exhibit topographical variation. PMID- 9416488 TI - Osteoid and bone formation in a nasal mucosal melanoma and its metastasis. AB - AIMS: To present a literature review and a case history concerning bone and osteoid formation by a metastasizing (mucosal) melanoma. CASE DETAILS: Osteocartilaginous differentiation and production of osteocartilaginous structures in malignant melanoma have been described only in 12 previous cases (osteoid in 11. bone in four), all of which involved dermal melanomas. Five of these melanomas were recurrent and one was associated with neurofibromatosis. The case report concerns a 75-year-old man with a nasal mucosal melanoma which was treated surgically. One year later, the patient developed a local recurrence and a cervical lymph node metastasis. Both the recurrent tumour and the metastasis showed clear evidence of bone and osteoid formation. CONCLUSIONS: This case is the first report in the literature, clearly demonstrating bone and osteoid formation by a mucosal melanoma, not only at the primary site, but even more convincingly in a cervical lymph node metastasis. PMID- 9416487 TI - Rheumatic Aschoff nodules revisited. II: Cytokine expression corroborates recently proposed sequential stages. AB - AIMS: A recent immunohistochemical analysis of the Aschoff lesions in rheumatic fever, combining immunohistochemical analysis with comparative morphology, permitted the division of the Aschoff nodules into three stages: (1) Aschoff nodule without admixed lymphocytes, (2) Aschoff nodules with a few T lymphocytes, and (3) Aschoff nodules containing many admixed lymphocytes of both B- and T-cell phenotype. It was postulated that the order of progression was from stage 1 with macrophages only, to accumulation of first T lymphocytes (stage 2) and then B lymphocytes (stage 3). This study was undertaken to determine the role and distribution of interleukin 1 (IL-1), interleukin 2 (IL-2) and tumour necrosis factor alpha (TNF alpha) in the various stages of the rheumatic Aschoff nodule to investigate our hypothesis on the progression of these nodules. METHODS AND RESULTS: Sixteen fresh valve specimens from patients with acute rheumatic fever undergoing valve surgery were obtained. Tissue sections from 14 specimens identified as containing Aschoff nodules were subjected to immunohistochemistry for (1) T and B lymphocytes, to stage the lesions according to our previously proposed criteria; (2) IL-1, IL-2 and TNF alpha; and (3) CD4 and CD8 to phenotype the T lymphocytes. The stage 1 and 2 lesions expressed IL-1 and TNF alpha in the macrophages. The stage 3 lesions showed more variable expression of all three cytokines including IL-2 within T lymphocytes. CONCLUSION: TNF alpha and IL-1 secretion in macrophages is required for T and B lymphocytes activation and aggregation; suggesting that macrophages arrive at the scene of rheumatic injury prior to the lymphocytes. IL-2 is usually expressed later in the inflammatory process and was found only in the lymphoid aggregates. This study therefore produces corroborative evidence for our previously proposed developmental stages of the Aschoff nodule. PMID- 9416489 TI - Primary alveolar soft-part sarcoma of the mediastinum: a clinicopathological and immunohistochemical study of two cases. AB - AIMS: Primary alveolar soft part sarcomas originating within the thoracic cavity are rare. The cases herein described highlight the ubiquitous distribution of this neoplasm and the importance of considering this tumour in the differential diagnosis of mediastinal tumours. METHODS AND RESULTS: Two cases of alveolar soft part sarcoma of the mediastinum are presented. The patients are two men of 22 and 23 years of age. Clinically, the patients presented with symptoms of chest pain. One tumour was in the anterior mediastinum while the second tumour was in the posterior mediastinum. Even though the bulk of the tumours were in mediastinal locations, both patients had pulmonary metastases at the time of diagnosis. Histologically, both tumours showed the classic morphology of alveolar soft part sarcoma, i.e. a proliferation of large polygonal cells with round to oval nuclei, prominent nucleoli and moderate amounts of eosinophilic cytoplasm arranged in a prominent alveolar pattern. Periodic acid-Schiff stains showed the characteristic diastase-resistant intracytoplasmic crystals. Immunohistochemical studies showed focal myoglobin reactivity in one case, while cytokeratin, vimentin, S100 protein, chromogranin, synaptophysin, neurofilaments, leu-enkephalin, desmin, smooth muscle actin and muscle-specific actin were negative in both cases. The patient with the anterior mediastinal tumour died of disease 14 months after diagnosis, and the patient with the posterior mediastinal mass remained well for at least 15 years and was later lost to follow-up. CONCLUSIONS: As has been observed in other anatomic areas, namely soft tissues, alveolar soft part sarcomas may follow an uncertain natural history. Interestingly, in our cases, the tumour in the anterior mediastinum followed a fatal course raising the possibility that the anatomic location of the tumour may have play a role in the behaviour of the tumour. PMID- 9416490 TI - Rhabdomyomatosis of the newborn lung unassociated with other malformations. AB - AIMS: To describe a case of rhabdomyomatosis of the lung unassociated with other external or visceral malformations in a newborn infant. METHODS AND RESULTS: A 26 weeks gestation newborn male with no relevant medical or family history presented a well-circumscribed solid area in the posterior mediastinum occupying the upper lobe of the right lung. The possibility of neuroblastoma or an extralobar pulmonary sequestration were excluded after laboratory and arteriographic studies. No visceral anomalies were found. At the age of 9 months the patient underwent a partial lobectomy, and he is free of disease 39 months after surgery. Histological examination demonstrated the presence of numerous bundles of striated fibres arranged haphazardly in the pulmonary interstitium in a background of a type-II congenital cystic adenomatoid malformation-like morphology of the resected lung. CONCLUSION: The presence of striated muscle fibres in the lung not necessarily represents a lethal congenital malformation. As this case shows, rhabdomyomatosis of the lung can affect a single pulmonary lobe, and resection of the affected lung parenchyma may be curative. It is important for pathologists to be aware of this entity, although it is exceptional, and to include it in the differential diagnosis of pulmonary masses in the newborn lung. PMID- 9416491 TI - Multiple ductal lipophagic polyps of the breast: a rare finding in mammary duct ectasia. PMID- 9416492 TI - Dermatofibroma with granular cells. PMID- 9416493 TI - Morphological changes in Pseudomonas aeruginosa secondary to ciprofloxacin. PMID- 9416494 TI - Rosai-Dorfman disease of the breast. PMID- 9416495 TI - Heterotopic ossification following biopsy--a sheep in wolf's clothing. PMID- 9416496 TI - Metanephric adenoma of the kidney: a rare benign tumour of the kidney. PMID- 9416497 TI - Rosenthal fibre formation in nervous tissues of benign ovarian cystic teratomas- a study of immunoreactivity for glial fibrillary acidic protein. PMID- 9416498 TI - Intraductal papilloma associated with metaplastic carcinoma of the breast. PMID- 9416499 TI - What can we learn about human immunodeficiency virus infection from a study of lymphocytic choriomeningitis virus? AB - The role of cytotoxic T lymphocytes (CTL) in human immunodeficiency virus (HIV) infection remains elusive. Since the discovery 10 years ago of high levels of specific CTL in this disease, some have argued that they play an important role in virus control, others that they drive disease progression through destruction of T helper cells, and others still that they play no obvious role at all. By contrast, the central role of CTL in murine lymphocytic choriomeningitis virus (LCMV) infection has been very clearly worked out through the use of in vivo depletion and adoptive transfer experiments, as well as knockout and transgenic mice. To interpret the possible roles for CTL in HIV, we have therefore made a comparison between what is known about CTL and their interaction with virus infected cells in these two infections. This illustrates a potential critical role for these cells in both control of HIV replication and immune-mediated pathology, but one that is highly dependent on virus dose, distribution and dynamics. PMID- 9416500 TI - Co-evolution of human immunodeficiency virus and cytotoxic T-lymphocyte responses. AB - After more than a decade of intensive research, the precise role of human immunodeficiency virus (HIV)-specific cytotoxic T lymphocytes (CTL) in determining the course of the infection remains open to argument. It is established that HIV-specific CTL appear early in the infection and are temporally associated with the clearance of culturable virus from the blood; that CTL are generally detectable at very high levels throughout the asymptomatic phase and decline at the time of progression to AIDS; and that CTL-mediated killing is sufficiently fast to prevent production of new virions by HIV-infected cells. However, viral turnover is high throughout the course of the infection, and infected individuals progress inexorably to disease in spite of the CTL response. In order to address the question of whether CTL play an active part in influencing the course of HIV infection, one approach has been to seek evidence for CTL-mediated selection pressure on the virus. Several clear examples of CTL epitope-specific mutations selected to fixation are described. We argue that CTL escape is a common event which occurs at all stages of the infection. Detailed longitudinal studies are required to detect CTL escape and to understand the complexities contributed by factors such as a polyvalent CTL response and the presence of epitope variants which antagonise the CTL response. In conclusion, there is strong evidence of a dynamic process in which CTL impose important selection constraints upon HIV from which the virus attempts to escape; ultimately, at the time of disease progression, the tenuous control of CTL over the virus is lost. PMID- 9416501 TI - Host factors in the pathogenesis of HIV disease. AB - Host factors play an important role in determining rates of disease progression in human immunodeficiency virus (HIV)-infected individuals. HIV is able to subvert the host immune system by infecting CD4+ T cells that normally orchestrate immune responses and by inducing the secretion of proinflammatory cytokines that the virus can utilize to its own replicative advantage. The recognition that certain chemokine receptors serve as necessary co-factors for HIV entry into its target cells as well as the fact that ligands for these receptors can modulate the efficiency of HIV infection has expanded the number and scope of host factors that may impact the pathogenesis of HIV disease. This area of investigation will no doubt yield novel therapeutic strategies for intervention in HIV disease; however, caution is warranted in light of the enormous complexity of the pleiotropic cytokine and chemokine networks and the uncertainty inherent in manipulating these systems. HIV-infected long-term non progressors represent an excellent model to study potential host factors involved in HIV disease pathogenesis. Genetic factors certainly have a major impact on the immune responses mounted by the host. In this regard, a polymorphism in the gene for the HIV co-receptor CC chemokine receptor 5 (CCR5), which serves as a co receptor for macrophage (M)-tropic strains of HIV, affords a high degree of protection against HIV infection in individuals homozygous for the genetic defect and some degree of protection against disease progression in HIV-infected heterozygotes. HIV-specific immune responses, including cytotoxic T-lymphocyte (CTL) responses and neutralizing antibody responses, also appear to play salutary roles in protecting against disease progression. PMID- 9416502 TI - Complement receptors in HIV infection. AB - The complement system plays an important role in the antimicrobial defense of the organism. Its components recognize a large variety of pathogens and target them for destruction, either directly by formation of a membrane attack complex or indirectly by recruiting phagocytic cells. In addition, it has several functions in cell activation, clearance of immune complexes, control of inflammatory reactions, chemotaxis and autoimmunity. For mediation of all these tasks of the complement system, complement receptor molecules on the cell surface play a key role. Current knowledge on structure, function, signal transduction and associated molecules is briefly summarized here. The role of complement receptors for human immunodeficiency virus (HIV)-associated pathogenesis is ambiguous and varies depending on cell type. On the one hand, complement receptors support the infected host to manage HIV infection and to defend itself, at least partially, against viral spreading throughout the organism. Such complement receptor mediated supporting mechanisms are activation of immune cells and lysis of viral particles and infected host cells. On the other hand, HIV employs complement receptors to intrude more easily into various cell types, to become localized into lymph follicles and to activate viral replication in latently infected cells. This review summarizes the complex interaction of virus and complement receptors in HIV infection for different cell types. PMID- 9416503 TI - Complement and immunity to viruses. AB - Complement is one of the first lines of host defence to be faced and countered by viruses as they struggle to establish an infection. As an important arm of the humoral immune response, the complement system is immediately ready to target and eliminate virus particles and to interact with the surface of virus-infected cells to mark them for destruction by other branches of the immune response. Nevertheless, some viruses are still very successful human pathogens. This article will discuss the role of complement in antiviral immunity, the mechanisms by which complement may be activated by viruses or virus-infected cells, and explore some of the strategies which viruses have evolved to subvert the immune response, including mechanisms by which complement activation may be prevented or aborted. PMID- 9416504 TI - Alpha beta and gamma delta T-cell networks and their roles in natural resistance to viral infections. AB - Both alpha beta and gamma delta T-cell populations and natural killer (NK) cells include cytotoxic, interferon (IFN)-gamma-producing lymphocytes that actively respond to viral infections. We show here that all three populations can provide "natural resistance" to viruses very early in infection and describe how the T cell populations are modulated to provide this function. gamma delta T cells were shown to play a role in controlling vaccinia virus (VV) infections, as VV grew to much higher titers in gamma delta T-cell knockout mice than in normal mice 3-4 days post-infection. Our studies of the alpha beta T-cell responses to viruses revealed an interactive network of T cells that is modulated substantially during systemic infections. There is an induction phase associated with a massive virus specific CD8 T-cell response, an apoptosis phase during which the T cells become sensitized to activation-induced cell death (AICD), a silencing phase, during which the T-cell number and activation state is reduced, and, finally, a memory phase associated with the very stable preservation of virus-specific memory cytotoxic T-lymphocyte precursors (pCTL). Infection of mice immune to one virus with a heterologous virus leads to a selective expansion of memory CTL cross reacting between the two viruses, but, after homeostasis is again established, there is a quantitative reduction and qualitative alteration of memory to the first virus. Our results suggest that memory alpha beta T cells cross-reactive between heterologous viruses mediate both immunopathology and protective immunity at early stages of the second virus infection. Thus, memory alpha beta T cells can, like gamma delta T cells and NK cells, provide natural immunity to viral infections. PMID- 9416505 TI - Role of the B-cell response in recovery of mice from primary influenza virus infection. AB - Recovery from influenza virus infection has long been known to require an intact T-cell compartment. More recent studies revealed that CD8 and CD4 T cells can promote recovery through independent mechanisms. The CD4 T-cell-dependent recovery process appears to operate primarily through promotion of the T dependent antibody response as B-cell-deficient microMT mice cannot recover from infection if they have been depleted of CD8 T cells. The potential therapeutic activity of the B-cell response was further studied by transfer of antibodies into infected SCID mice. At the dose of 200 micrograms/mouse, most antibodies (of IgG2a isotype) to the viral transmembrane protein HA cured the infection, while those to the transmembrane proteins NA and M2 suppressed virus titers in the lung but failed to clear the infection. The ability of passive antibody to resolve the infection was closely related to its prophylactic activity, suggesting that neutralization of progeny virus (VN) played an important role in the process of virus clearance in vivo, while reaction of antibodies with infected host cells contributed to but was insufficient, on its own, for cure. HA-specific antibodies of IgM and IgA isotypes were therapeutically ineffective against pulmonary infection, presumably because of a preferential delivery into the upper respiratory tract, while IgG exhibited highest activity against pulmonary and minimal activity against nasal infection. B cells appear to be of similar importance for recovery from primary infection as CD8 T cells. PMID- 9416506 TI - Effector CD4+ and CD8+ T-cell mechanisms in the control of respiratory virus infections. AB - The rules for T-cell-mediated control of viruses that infect via the respiratory mucosae show both common themes and differences, depending on the nature of the pathogen. Virus-specific CD8+ cytotoxic T lymphocytes (CTLs) are the key effectors of virus clearance in mice infected with both negative strand RNA viruses (influenza and Sendai) and a DNA virus, the murine gamma-herpesvirus-68 (MHV-68). Recently completed experiments establish that these activated CD8+ T cells indeed operate primarily via contact-dependent lysis. Perforin-mediated cytotoxicity seems to be the preferred mode, though a Fas-based mechanism can apparently serve as an alternative mechanism. Immune CD4+ T cells functioning in the absence of the CD8+ subset cannot eliminate MHV-68 from lung epithelial cells, are somewhat less efficient than the CD8+ CTLs at clearing the RNA viruses, and are generally ineffectual in mice that lack B lymphocytes. Though cytokine secretion by CD4+ and CD8+ T cells in the virus-infected lung may promote both T-cell extravasation and macrophage activation, such processes are not alone sufficient to deal consistently with any of these infections. However, CD4+ T help is mandatory for an effective B-cell response, and can operate to promote the clonal expansion of virus-specific CD8+ T cells in the lymph nodes and spleen. Furthermore, a concurrent CD4+ T-cell response seems to be essential for maintaining continued CD8+ T-cell surveillance and effector capacity through the persistent, latent phase of MHV-68 infection in B cells. Thus, the evidence to date supports a very traditional view; CD8+ T cells function mainly as killers and the CD4+ T cells as helpers in these respiratory virus infections. PMID- 9416507 TI - Cytokines and immunity to viral infections. AB - In this review, we discuss two broad approaches we have taken to study the role of cytokines and chemokines in antiviral immunity. Firstly, recombinant vaccinia viruses were engineered to express genes encoding cytokines and chemokines of interest. Potent antiviral activity was mediated by many of these encoded factors, including IL-2, IL-12, IFN-gamma, TNF-alpha, CD40L, Mig and Crg-2. In some cases, host defense mechanisms were induced (IL-2, IL-12, Mig and Crg-2), whilst for others, a direct antiviral effect was demonstrated (IFN-gamma, TNF alpha and CD40L). In sharp contrast, vector-directed expression of IL-4, a type 2 factor, greatly increased virus virulence, due to a downregulation of host type 1 immune responses. Our second experimental approach involved the use of strains of mice deficient for the production of particular cytokines or their receptors, often in combination with our engineered viruses. Mice deficient in either IFN gamma, IFN-gamma R, IFN-alpha/beta R, TNFRs, CD40 or IL-6 were, in general, highly susceptible to poxvirus infection. Surprisingly, not only the TNFR1, but also the TNFR2, was able to mediate the antiviral effects of TNF-alpha in vivo, whilst the antiviral activity observed following CD40-CD40L interaction is a newly defined function which may involve apoptosis of infected cells. Through the use of perforin-deficient mice, we were able to demonstrate a requirement for this molecule in the clearance of some viruses, such as ectromelia virus, whilst for others, such as vaccinia virus, perforin was less important but IFN-gamma was essential. PMID- 9416508 TI - Vaccinia virus immune evasion. AB - Vaccinia virus and other poxviruses express a wide variety of proteins which are non-essential for virus replication in culture but help the virus to evade the host response to infection. Examples include proteins which oppose apoptosis, synthesise steroids, capture chemokines, counteract complement, interfere with interferon and intercept interleukins. This review provides an overview of such proteins, with an emphasis on work from our laboratory, and illustrates how the study of these proteins can increase our understanding of virus pathogenesis, the function of the immune system and how to make safer and more immunogenic poxvirus based vaccines. PMID- 9416509 TI - The role of antibody in recovery from alphavirus encephalitis. AB - Alphaviruses infect neurons in the brain and spinal cord and cause acute encephalomyelitis in a variety of mammals. The outcome of infection is determined by whether the neurons survive infection and this, in turn, is determined by the virulence of the virus and the age of the host at the time of infection. We have been studying Sindbis virus (SV) infection of mice as a model system for alphavirus-induced encephalomyelitis. Investigation of intracerebral infection of weanling mice with two different strains of SV has allowed us to analyze the role of the immune response in protection from fatal disease (virulent NSV strain) and in clearance of virus from the nervous system during non-fatal disease (less virulent SV AR339 strain). Neutralizing and non-neutralizing antibodies to the E1 and E2 surface glycoproteins can protect mice from fatal NSV infection when given before or after infection, while T cells are not protective. The mechanism of antibody-mediated protection is not known, but it is likely that more than one mechanism is involved and that different mechanisms are involved in pre-infection and post-infection treatment protection. Clearance of infectious virus from the nervous system of mice during recovery from non-fatal disease is accomplished by antibodies to the E2 glycoprotein. The process does not involve damage to the infected neurons and is independent of complement and mononuclear cells. Bivalent antibody is required and binds to the surface of the infected cell. Initially, release of virus by budding from the cell surface is prevented and, subsequently, intracellular virus replication is inhibited possibly through antiviral mechanisms induced in co-operation with interferon. This non-lytic mechanism for control of virus infection results in the prolonged presence of viral RNA in tissue and the need for prolonged intrathecal synthesis of antiviral antibody by B cells within the central nervous system. PMID- 9416510 TI - The infection of mouse by Theiler's virus: from genetics to immunology. AB - Theiler's virus is a picornavirus of mouse which causes an acute encephalomyelitis followed by a persistent infection of the white matter of the spinal cord with chronic inflammation and demyelination. This late disease is studied as a model for multiple sclerosis. Inbred strains of mice differ in their susceptibility to persistent infection and demyelination. Resistant strains clear the infection after the acute encephalomyelitis. This observation is the basis of genetic studies which we used as a thread for this review. The H-2D locus has a major effect on susceptibility. The H-2Db gene is involved in a fast and intense CTL response which confers resistance. The Tcrb locus is also implicated, although there is no proof that the susceptibility gene in this region codes for the T-cell receptor. A complete screen of the genome uncovered the role of the Ifng locus and led to the demonstration that IFN-gamma limits viral spread in the white matter. The roles of NK cells and B cells in limiting the infection are discussed. CD4+ T cells participate both in protection against the infection and in demyelination. Finally, the effect of non-immune factors in resistance is illustrated by mice with mutations in the MBP or PLP gene. PMID- 9416511 TI - Antiviral immune responses modulate the nature of central nervous system (CNS) disease in a murine model of multiple sclerosis. AB - The spectrum of disease is influenced by factors related to both the pathogen and the host, as well as the end points used in defining disease. In this article, the issue of disease resistance versus susceptibility will be examined in the framework that genetic manipulation of either the pathogen or the host immune response alters the balance from disease protection towards pathogenesis. The response of the host may trigger both a protective and a pathogenic immune response. The failure to mount a protective immune response predisposes the pathogen to persistence, which then becomes the target for immunopathology. This review will examine the factors involved both in virus-mediated pathogenesis and in disease protection in the Theiler's model of human multiple sclerosis. By manipulating the character of the virus pathogen and the specificity of the immune response, the entire spectrum of human demyelinating disease is reproduced. PMID- 9416512 TI - On the mystique of the immunological self. AB - Since the time of Paul Ehrlich 100 years ago, we have known that the immunological apparatus somehow inhibits most damaging autoimmune responses while permitting a response to exogenous immunogens. With the discovery of tolerance, the concept of immunological surveillance, and especially with the discovery of HLA restriction of T-cell recognition, the term "the immunological self" and the phrase "self-nonself discrimination" have gained wide currency. Immunology has been called "The Science of Self", and self-nonself discrimination has been assigned as the driving force for its complex evolution. The concept of self has thus been given such mystical trappings since the time of Macfarlane Burnet that recent workers have felt free to pronounce it the central paradigm of modern immunology, and to claim to overthrow it! In this article, we challenge some of the more egregious claims about the immunological self by recalling important historical findings, by reviewing the mechanisms of Darwinian evolution, and by remembering that the general pathology of immunogenic inflammation shows that the immune response cannot discriminate between the benign and the noxious. PMID- 9416513 TI - The antigenic potentiation of inhalant allergens by bacterial compounds in bronchial asthma. PMID- 9416514 TI - Cytokines and asthma. PMID- 9416515 TI - Adhesion molecules in allergy. PMID- 9416516 TI - The role of bronchial epithelium in asthma. PMID- 9416517 TI - Role of leukotrienes and leukotriene receptor antagonist in asthma: new advances. PMID- 9416518 TI - Improvement of ventilatory function in asthmatic children after respiratory rehabilitation. PMID- 9416519 TI - Utility of the study of non-specific bronchial response for monitoring bronchial asthma. PMID- 9416520 TI - The role of airway resistance in the diagnosis of bronchial hyperreactivity. PMID- 9416521 TI - Evaluation of compliance with prescribed treatment in asthmatic children and adolescents: a Portuguese problem. PMID- 9416522 TI - I.S.A.A.C. World Project: epidemiology in the Cape Verde and Madeira Islands. PMID- 9416523 TI - Impact of asthma in a Spanish health area. PMID- 9416524 TI - Epidemiology of asthma in the Iberian Peninsula. PMID- 9416525 TI - Epidemiological aspects of asthma in the Canary Islands. PMID- 9416526 TI - Epidemiological aspects of asthma in Latin America. PMID- 9416527 TI - Asthma in infants. PMID- 9416528 TI - Immune responses in early childhood in relation to the development of asthma. PMID- 9416529 TI - Early detection of chronic evolution of asthma in children. PMID- 9416530 TI - Asthma among the elderly. PMID- 9416531 TI - Diagnostic basis of occupational asthma. PMID- 9416532 TI - Occupational asthma as an etiopathogenic model for asthma induced by common allergens. PMID- 9416533 TI - Asthma due to inhalation of foods. PMID- 9416534 TI - Latex hypersensitivity: a worldwide crisis. PMID- 9416535 TI - Asthma due to the ingestion of contaminated flour. PMID- 9416536 TI - Usefulness of specific bronchial challenge in monitoring immunotherapy in occupational asthma. PMID- 9416537 TI - Dehumidification and environmental control. PMID- 9416538 TI - The importance of tobacco in environmental control. PMID- 9416539 TI - Air cleaners and airborne allergens. PMID- 9416540 TI - In vitro diagnosis for bronchial asthma. PMID- 9416541 TI - A functional approach for the assessment of the components of airways obstruction in asthma. PMID- 9416543 TI - Non-invasive methods for diagnosing asthma. Study of induced sputum. PMID- 9416542 TI - Eosinophil cationic protein in bronchoalveolar lavage from wheezy infants. PMID- 9416544 TI - Biological standardization systems for allergenic extracts. PMID- 9416545 TI - The position of immunotherapy in the European Academy of Allergology and Clinical Immunology. PMID- 9416546 TI - The safety and efficiency of immunotherapy. PMID- 9416547 TI - Novel approaches to immunotherapy: epitopes, determinants, activators, or modulators? PMID- 9416548 TI - Costs of specific immunotherapy. PMID- 9416549 TI - Sublingual immunotherapy: accumulated experience. PMID- 9416550 TI - Preventive allergy treatment as part of allergy disease management. PMID- 9416551 TI - Capacity of specific immunotherapy in prevention of allergic asthma in children: the Preventive Allergy Treatment Study (PAT). PMID- 9416552 TI - Long-lasting beta 2-agonists in the treatment of asthma. PMID- 9416553 TI - The prevention of occupational asthma in industries. AB - This paper discusses criteria for the primary and secondary prevention of occupational asthma. The former involves intervention in the workplace to prevent onset of the disease, while the latter is based on early diagnosis. These forms of prevention are applied in this study, particularly to work involving laboratory animals, the use of latex gloves, wheat flour, proteolytic enzymes and simple chemical compounds (isocyanates). PMID- 9416554 TI - Current trends in treatment using theophylline. PMID- 9416555 TI - Pharmacology of fluticasone propionate. AB - Fluticasone propionate is a novel inhaled corticosteroid which has the interesting properties of a high activity combined with low gastrointestinal absorption and rapid liver metabolism. On theoretical grounds, this drug might be expected to serve as a good antiinflammatory agent in asthma, while at the same time causing minimal systemic side effects. PMID- 9416556 TI - Role of cyclooxygenase 1 and 2 in asthma and rhinitis. PMID- 9416557 TI - Cough syndrome. Reflux pharyngitis. PMID- 9416558 TI - The management of ASA syndrome. PMID- 9416559 TI - Rhinitis and asthma: nasal provocation test in the diagnosis of asthma. AB - Our purpose was to determine if the study of rhinitis is useful in the diagnosis of asthma. We formulated the hypothesis that the inflammation of the upper airway reflects the inflammation of the lower airway. It was found that there are allergens that produce rhinitis more frequently than asthma, and vice versa. This can be explained by size. This explanation, however, is questionable as the allergic proteins are extracted from the carrying agent, and through the lymphatic route or the blood, reach the entire human organism. It was also found that with bronchoalveolar lavage in allergic asthma it is possible to obtain the same results for eosinophils as with a nasal wash or using Citospyn. However, the results in the late phase are questionable. In the immediate phase and in the late phase, eosinophil cationic protein (ECP) was detected in the blood (in asthma) and in nasal washes (in rhinitis). In the immediate response tryptase was detected from the mast cells. The role of leukotrienes in asthma and rhinitis is well established in the early and late response. The use of leukotriene inhibitors guarantee their importance in the airway. Platelet-activating factor (PAF) has been demonstrated to increase vascular permeability and the use of antagonists were the best nasal feature. The inhalation of histamine caused bronchospasm, while instillation of histamine in the nasal passages increased resistance. With this information it seems that our hypothesis has been confirmed. Rhinitis and BHR together are equivalent to asthma, although the PFER decreases during the course of nasal provocation test (NPT) in nonasthmatic patients. In pure rhinitis patients, however, we find decreases in PFER due to effort. All this suggests that the study of nasal inflammation is still unclear with regard to bronchial inflammation. PMID- 9416560 TI - Allergenicity of Blomia tropicalis. PMID- 9416561 TI - Allergen properties of Lepidoglyphus destructor. PMID- 9416562 TI - Occupational asthma from storage mites contaminating foods. PMID- 9416563 TI - Prevalence of sensitivity to storage mites in our surroundings. PMID- 9416564 TI - Importance of sensitivity to storage mites in bronchial asthma. PMID- 9416565 TI - Inflammatory actions of platelet-activating factor: control by PAF acetylhydrolase. PMID- 9416566 TI - International development of portable inhalers. PMID- 9416567 TI - Workshop on respiratory rehabilitation in patients with bronchial asthma. PMID- 9416568 TI - Immune stimulation may contribute to enhanced progression of SIV induced disease in rhesus macaques. AB - A number of rhesus macaques experimentally infected with SIV isolates such as SIVmac251, fail to seroconvert, develop high plasma viremia and die rapidly (within 6-7 months p.i.). We hypothesized that such rapid progression is a result of a state of hyperimmune activation and concomitant immune suppression of these animals at the time of virus challenge. In efforts to test the hypothesis that immune activation leads to rapid progression of lentivirus-induced disease, adult rhesus macaques were infected with SIV mac251 and received an alternate monthly schedule of repeated immunization with allogeneic cells, keyhole limpet hemocyanin and tetanus toxoid (group I). For purposes of controls, a group of monkeys was infected with the same pool and dose of virus but were not immunized (group II) and a group was immunized with the same schedule of multiple antigens as group I but were not infected with SIV (group III). All the animals in group I (n = 3) either failed to seroconvert or developed very low levels of SIV antibodies, had high plasma p27 defined antigenemia, and died within 8 months (2/3 died within 4 months). Of the animals in group II (n = 8), two patterns emerged as we had noted before. One subgroup (3 animals), displayed the same profile as group I (failure to fully seroconvert, high p27 levels and death by 8 months), whereas the other subgroup (5 animals) seroconverted, had low plasma p27 levels, and survived past 11 months (2/5 still alive past 22 months). All 3 animals in group III remained healthy. The data provided herein suggest that either experimental or natural (due to factors not clear at present) immune stimulation may lead to accelerated lentivirus induced disease progression most likely due to immune suppression and has implications for the understanding of the mechanisms for the rate of disease progression in human HIV-1 infection. PMID- 9416569 TI - Effects of chair-restraint on gastrointestinal transit time and colonic fermentation in male rhesus monkey (Macaca mulatta). AB - The incidence of an 18 day chair-restraint on the digestive physiology of male rhesus monkey was investigated for space research purposes, comparing four trained restraint subjects with two vivarium controls. Chair-restraint induced a 2.5-fold acceleration of the gastrointestinal transit time, which persisted throughout the 7 day postrestraint period, and an increase of the fecal dry matter content, which mean value rose from 40.7% to 69.6%. Fecal pH remained unaltered throughout the experiment. Modifications of fermentative metabolites produced by the colonic microflora and excreted through the breath (hydrogen and methane) or in the feces (short chain fatty acids and ammonia) could not be reliably related to chair-restraint and probably involved side-stress factors. On the whole, alterations due to chair-restraint are shown to be different from those reported in the literature, following a modification of the dietary composition. These data may help to predict the alterations of digestive physiology likely to occur in immobilized human patients. PMID- 9416570 TI - The brachial plexus in the chacma baboon (Papio ursinus). AB - The brachial plexus in each of ten embalmed, mature chacma baboons was dissected to document the structure and branching pattern of this nerve plexus in this increasingly used research animal. In general, the brachial plexus in the chacma baboon was similar to the plexuses in the vervet and other Old World monkeys. However, several aspects were comparable to those observed in domestic animals. Thus the bipedal and quadrupedal abilities of the chacma baboon were reflected in the structure of its brachial plexus. PMID- 9416571 TI - Prevalence of antibody to Plasmodium falciparum antigens among feral Saimiri monkeys in the Amazon basin region of Peru. PMID- 9416572 TI - Proteinuric hypertension in a pregnant baboon: was this pre-eclampsia? AB - We report the results of investigating a pregnant baboon that developed hypertension, proteinuria, and oedema in late gestation. Although the clinical presentation suggested a diagnosis of pre-eclampsia, the evolution of her clinical signs and results of a renal biopsy performed 3 weeks after delivery suggested that glomerulonephritis was the underlying disease. PMID- 9416573 TI - Oral shedding of herpes simplex virus type 1: a review. AB - Herpes simplex virus type 1 (HSV-1) and, to a lesser extent, type 2 (HSV-2) are the aetiological agents of recrudescent herpes labialis (RHL). The available literature on patterns of HSV-1 shedding into the oral cavity at the prodromal stage of disease, during recrudescences and also during asymptomatic periods, is reviewed, as are the potential sources of virus and the known trigger factors leading to viral reactivation. Attention is given to the methodologies in use for the detection of HSV-1 and the relevance to the risk of cross-infection in surgery. This review also discusses the increase in incidence of HSV-1 genital infections and the significance of salivary inhibitors of the herpes simplex type 1 virus. PMID- 9416574 TI - Depressive symptoms in individuals with idiopathic subjective dry mouth. AB - It has been known for many centuries that there is a relationship between saliva flow rate and emotional status. The significance of psychological processes in the subjective sensation of a dry mouth has been discussed earlier, and this study deals with the presence of depressive symptoms in individuals with idiopathic subjective sensation of a dry mouth. Depressive symptoms in 94 healthy subjects with normal flow rates for unstimulated and stimulated whole saliva but with a subjective sensation of a dry mouth were assessed by the Beck Depression Inventory (BDI) and compared with healthy age- and gender-matched controls. The subjects with a subjective dry mouth condition were significantly more depressive and also had a significantly higher frequency of depressive symptoms. Depression was found in 21.3% of the individuals with a subjective dry mouth sensation and in 3.2% of the controls. The results of this study indicate that, in some cases, subjective dry mouth may be of psychological origin. PMID- 9416575 TI - Immunochemical analysis of laminin in duct-ligated submandibular glands of rats. AB - Immunochemical methods have been used to study the expression of laminin during experimental atrophy of the submandibular gland of rats caused by ductal ligation. In normal submandibular glands, laminin immunoreactivity appeared as continuous linear staining around acini and ducts. In the ligated glands, it exhibited an irregular pattern and intensity. Staining was usually stronger around small ducts and acini, which were most prominent in glands ligated for 30 days. Immunoblot analysis showed that the laminin of the rat submandibular gland contains bands that correspond to the EHS alpha 1, beta 1 and gamma 1 chains, and that the composition of the laminin chains does not change during the atrophy. PMID- 9416576 TI - Absence of Helicobacter pylori DNA in salivary lymphoepithelial lesions. AB - Helicobacter pylori is a common cause of chronic gastritis and has been implicated as the main agent responsible for the development of lymphomas of mucosa associated lymphoid tissue (MALT) in the stomach. An uncommon cause of salivary gland swelling is salivary lymphoepithelial lesion (SLEL), which shows histological features of acquired MALT and is associated with the development of MALT-type lymphomas. Since H. pylori has been identified in the oral cavity, we hypothesised that this organism might act as a potential antigen for the development of MALT in salivary glands. Routinely processed biopsies of 20 SLEL were screened for H. pylori DNA using a sensitive two-stage PCR technique to amplify the 16S ribosomal RNA gene. Immunoglobulin heavy chain gene monoclonality was determined by amplifying the VDJ gene using a nested PCR technique. All SLEL had histological features of organised MALT and 14 cases showed Ig heavy chain gene monoclonality consistent with MALT lymphoma. None of the SLEL contained H. pylori DNA. In contrast to the putative role of H. pylori as an antigenic stimulus in gastric MALT lymphomas, it appears not to play a role locally in the development of MALT or MALT lymphomas of the salivary gland. PMID- 9416577 TI - Immunolocalization of interstitial collagenase (MMP-1) and tissue inhibitor of metalloproteinases-1 (TIMP-1) in radicular cysts. AB - To investigate the mechanisms involved in expansion of radicular cysts, monoclonal antibodies against interstitial collagenase (MMP-1) and tissue inhibitor of metalloproteinases-1 (TIMP-1) were used to localize the sites of MMP 1 and TIMP-1 expression in 30 radicular cysts. Positive MMP-1 staining was detected in the lining epithelium and subepithelial fibroblasts, macrophages, endothelial cells and osteoblasts/osteocytes in all specimens. Positive TIMP-1 staining was identified in osteoblasts/osteocytes and endothelial cells of all specimens, and in the lining epithelium and subepithelial fibrous connective tissue wall of five radicular cysts with an intense inflammatory cell infiltrate. The number and distribution of positive cells for MMP-1 or TIMP-1 varied widely among individual specimens, but strong immunostaining was constantly detected at sites with prominent subepithelial inflammation. Results here support the hypothesis that MMP-1 may play an important role in the expansion of radicular cysts. The absence of TIMP-1 expression in lining epithelium and subepithelial fibroblasts and macrophages in most cases studied indicated that an imbalance between MMP-1 and TIMP-1 production may lead to radicular cyst expansion. PMID- 9416578 TI - Effect of non-erupted third molars on roots of approximal teeth. A radiographic, clinical and histologic study. AB - This study was undertaken to evaluate clinically and histologically root resorption in extracted human second molars in close proximity to non-erupted third molars. The control group consisted of extracted second molars that were proximal to fully erupted third molars. Eight out of the 11 teeth in the study group presented different degrees of radiographic root resorption, nine presented clinical resorption, and all 11 had histologic evidence of root resorption. In the control group, no signs of root resorption were seen radiographically or clinically. Histologically, limited sites of resorption were identified in all teeth, which were partially repaired by cellular cementum. Histologic observation of study specimens revealed root surface resorption in 10 out of the 11 teeth, one showing replacement resorption as well. Inflammatory resorption was observed in the three most advanced cases in the study group. Reparative cementum partially lining resorbed areas was evident in all teeth with surface resorption. Within the limits of this study, radiographic identification of distal root resorption of second molars in close proximity to non-erupted third molars appears reliable. The findings may support the hypothesis that the presence of a non-erupted third molar in close proximity to the distal root of the second results in root resorption. PMID- 9416580 TI - Discrimination of complex histopathological tumour profiles by experienced and inexperienced observers. AB - One of the tasks that pathologists routinely perform in diagnosis is the assessment of complexity of shape or texture of microscopic images. Because some of these complex patterns can be approximated in terms of fractal structures, it is important to understand how fractal structures are perceived by pathologists, and whether pathologists use specific perceptual strategies to quantify such patterns. Knowledge about this could be advantageous for either producing specific training programmes to increase complexity discrimination or to utilise more unbiased and quantitative imaging methodologies in histopathology. We have previously demonstrated that non-expert observers with high abilities in simultaneous information processing were best at the task of discriminating the fractal dimension of self-similar computer-generated curves. Here we compare the performance of experienced and inexperienced observers in a similar task of discriminating epithelial profiles. The results suggest that there are no differences between experienced and inexperienced observers in this task of complex profile discrimination. PMID- 9416579 TI - Differential expression of type I cytokeratins in hamster cheek pouch epithelium following treatment with dimethylbenzanthracene. AB - Cytokeratin (CK) expression in untreated, paraffin-treated or dimethylbenzanthracene (DMBA)-treated hamster cheek pouch epithelium was investigated utilizing monoclonal antibodies AE1 or AE3, which react with type I or type II CKs, respectively, and by in situ hybridization utilizing type I CK specific probes. The latter were isolated from a cDNA library of hamster cheek pouch mRNA and designated CK 13 and CK 10 based on their respective homologies (> 95% amino acids) with murine CK 13 and human CK 10. Treatment of hamster cheek pouch epithelium with DMBA resulted in increased expression of type I CK, detected immunohistochemically with monoclonal AE1, but decreased expression of type II CKs detected with AE3. Despite an overall increase in type I CKs, in situ hybridization demonstrated differential expression of type I CKs with altered distribution of CK 13 mRNA and reduced expression of CK 10 mRNA, providing additional sensitive markers for DMBA-associated changes in CKs. These changes were constant at 2 to 22 weeks in the pre-neoplastic and neoplastic epithelium following the initial application of DMBA. PMID- 9416581 TI - Carcinoma of the oral tongue in northern Finland: trends in overall incidence and patient and tumour characteristics. AB - A population-based survey was conducted in the two northernmost provinces of Finland to describe the incidence of tongue cancer as well as patient and tumour characteristics in cases diagnosed between 1974 and 1994. A total of 105 new patients with cancer of the oral tongue were included in the 21-year study period. The age-standardised incidence (per 100,000 years) of the carcinoma in men increased from 0.6 in the first 7-year period (1974-1980) to 1.0 in the last period (1988-1994). The incidences in women were 0.7 to 1.4, respectively. The average patient profile remained much the same through the years. The median duration of symptoms also remained the same over the 2 decades, as did the median size and location of the tumour at diagnosis. In conclusion, the incidence of carcinoma of the tongue about doubled in both the male and the female population from 1974 to 1994. However, the patient and tumour characteristics remained about the same, the tumours being relatively large at the time of diagnosis in spite of well-developed community health and dental care. PMID- 9416582 TI - Juvenile xanthogranuloma occurring in the oral cavity: case report and histopathological findings. AB - Juvenile xanthogranuloma is commonly seen in the dermis, and only very rarely develops in the oral mucosa. Here were report a case that occurred in the anterior palate of a 9-year-old boy. The lesion appeared as a dark red and well defined nodule measuring 12 x 14 mm. Histologically, it consisted of a proliferation of histiocytes and fibroblastic stroma intermingled with foamy cells. Many lipid droplets without limiting membrane were observed in the cytoplasm under electron microscopy, but no Langerhans' cell granules were observed. The proliferative histiocytes were positive for lysozyme and macrophage HAM56 under immunohistochemical observation, but not for S-100 protein. From these findings, the lesion was diagnosed as juvenile xanthogranuloma. The post operative course, now amounting to 7 years, has been uneventful. PMID- 9416583 TI - Vestibular autonomic regulation: overview and conclusions of a recent workshop at the University of Pittsburgh. PMID- 9416584 TI - Neuroanatomic substrates for vestibulo-autonomic interactions. AB - Recent anatomical studies have identified a network of central neural circuits that appear to integrate vestibular and autonomic information. Like vestibulo ocular and vestibulospinal circuits, these pathways appear to be under inhibitory modulation by distinct regions in the medial aspect of the cerebellar cortex. These central circuits have the potential to explain the known influence of vestibular stimulation on autonomic motor responses through descending effects on brain stem autonomic regions. In a more global context, the extensive convergence of vestibular and autonomic information in both vestibular and autonomic brain regions is consistent with the concept that vestibular and visceral information (for example, blood pooling and visceral proprioception) are used to form a central representation of gravitoinertial parameters during movements. This representation can influence neural circuitry involved in postural control, cardiovascular control, perception of the spatial vertical and emotional or affective responses. PMID- 9416585 TI - Physiological evidence that the vestibular system participates in autonomic and respiratory control. AB - Electrical or natural stimulation of the vestibular system results in changes in blood pressure and respiratory motor output. An increase in excitatory drive on the sympathetic nervous system occurs during nose-up vestibular stimulation in cats; this response is appropriate to offset orthostatic hypotension that could result from nose-up body rotations during movements such as vertical climbing. In addition, transection of the vestibular nerves in anesthetized or awake cats compromises the ability to correct decreases in blood pressure that result from nose-up body tilt. The vestibular system also has influences on respiratory muscles; these effects are appropriate to participate in making adjustments in the activity of respiratory muscles that are necessary to offset mechanical constraints on these muscles that occur during changes in body position. These data thus suggest that the influences of the vestibular system on the autonomic and respiratory systems serve to maintain homeostasis during movement. PMID- 9416586 TI - Clinical evidence that the vestibular system participates in autonomic control. AB - The vestibular system, including both the peripheral vestibular system, that is, the labyrinth, and the central vestibular system, is known to influence autonomic function in several ways that have clinical implications. This paper discusses evidence for vestibular influences on autonomic control from normal human subjects, evidence for vestibular influences on autonomic control from patients, clinical implications of vestibulo-autonomic regulation, and speculations regarding possible clinical implications of vestibulo-autonomic control. Situations that provoke vestibular-induced autonomic responses in normal subjects include vestibular laboratory testing, vehicular motion, simulators, and, possibly, exposure to microgravity. Patients with peripheral and central vestibular abnormalities manifest both symptoms and signs of autonomic dysfunction presumably via vestibulo-autonomic connections. Vestibulo-autonomic regulation impacts vestibular diagnostic testing, clinical diagnosis of balance disorders, and treatment of balance disorders. In addition to well-recognized peripheral and central vestibular disorders, anxiety disorders have recently been linked to vestibular dysfunction in a subset of patients. In particular, vestibular dysfunction has been linked to panic disorder and agoraphobia. Vestibular-autonomic connections may form a basis for an association between vestibular dysfunction and panic attacks. The importance of vestibulo-autonomic regulation in the clinical arena is not fully known. Two speculative areas discussed in this paper include vestibular-induced orthostatic intolerance and the role of vestibular-respiratory pathways on sleep apnea. PMID- 9416587 TI - Vestibular influences on autonomic cardiovascular control in humans. AB - There is substantial evidence that anatomical connections exist between vestibular and autonomic nuclei. Animal studies have shown functional interactions between the vestibular and autonomic systems. The nature of these interactions, however, is complex and has not been fully defined. Vestibular stimulation has been consistently found to reduce blood pressure in animals. Given the potential interaction between vestibular and autonomic pathways this finding could be explained by a reduction in sympathetic activity. However, rather than sympathetic inhibition, vestibular stimulation has consistently been shown to increase sympathetic outflow in cardiac and splanchnic vascular beds in most experimental models. Several clinical observations suggest that a link between vestibular and autonomic systems may also exist in humans. However, direct evidence for vestibular/autonomic interactions in humans is sparse. Motion sickness has been found to induce forearm vasodilation and reduce baroreflex gain, and head down neck flexion induces transient forearm and calf vasoconstriction. On the other hand, studies using optokinetic stimulation have found either very small, variable, or inconsistent changes in heart rate and blood pressure, despite substantial symptoms of motion sickness. Furthermore, caloric stimulation severe enough to produce nystagmus, dizziness, and nausea had no effect on sympathetic nerve activity measured directly with microneurography. No effect was observed on heart rate, blood pressure, or plasma norepinephrine. Several factors may explain the apparent discordance of these results, but more research is needed before we can define the potential importance of vestibular input to cardiovascular regulation and orthostatic tolerance in humans. PMID- 9416588 TI - Interaction of semicircular canal stimulation with carotid baroreceptor reflex control of heart rate. AB - The carotid-cardiac baroreflex contributes to the prediction of orthostatic tolerance; experimental attenuation of the reflex response leads to orthostatic hypotension in humans and animals. Anecdotal observations indicate that rotational head movements about the vertical axis of the body can also induce orthostatic bradycardia and hypotension through increased parasympathetic activity. We therefore measured the chronotropic response to carotid baroreceptor stimulation in 12 men during varying conditions of vestibulo-oculomotor stimulation to test the hypothesis that stimulation of the semicircular canals associated with head movements in the yaw plane inhibits cardioacceleration through a vagally mediated baroreflex. Carotid-cardiac baroreflex response was assessed by plotting R-R intervals (ms) at each of 8 neck pressure steps with their respective carotid distending pressures (mmHg). Calculated baroreflex gain (maximal slope of the stimulus-response relationship) was measured under 4 experimental conditions: 1) sinusoidal whole-body yaw rotation of the subject in the dark without visual fixation (combined vestibular-oculomotor stimulation); 2) yaw oscillation of the subject while tracking a small head-fixed light moving with the subject (vestibular stimulation without eye movements); 3) subject stationary while fixating on a small light oscillating in yaw at the same frequency, peak acceleration, and velocity as the chair (eye movements without vestibular stimulation); and 4) subject stationary in the dark (no eye or head motion). Head motion alone and with eye movement reduced baseline baroreflex responsiveness to the same stimulus by 30%. Inhibition of cardioacceleration during rotational head movements may have significant impact on functional performance in aerospace environments, particularly in high-performance aircraft pilots during high angular acceleration in aerial combat maneuvers or in astronauts upon return from spaceflight who already have attenuated baroreflex functions. PMID- 9416589 TI - Sensory conflict theory and space sickness: our changing perspective. PMID- 9416590 TI - The relative roles of the otolith organs and semicircular canals in producing space motion sickness. AB - Inflight and post-landing "immunity" to the "coriolis sickness susceptibility test", observed during the Skylab M131 experiment, suggests that the otolith organs play a major role in space motion sickness (SMS). This view is supported by the report that ocular counter-torsion asymmetries correlate with SMS incidence and severity. Further data indicate that sensory-motor adaptation to microgravity includes a process whereby central interpretation of otolith signals is biased from "tilt" toward translation. However, unexpected responses to linear acceleration suggest the importance of graviceptors distributed throughout the body in addition to the vestibular otolith organs. Research is needed to assess distributed graviceptor effects. PMID- 9416591 TI - Pharmacology of motion sickness. PMID- 9416592 TI - Managing space motion sickness. AB - Space motion sickness is a well-recognized problem for space flight and affects 73% of crewmembers on the first 2 or 3 days of their initial flight. Illness severity is variable, but over half of cases are categorized as moderate to severe. Management has included elimination of provocative activities and delay of critical performance-related procedures such as extra-vehicular activity (EVA) or Shuttle landing during the first three days of missions. Pharmacological treatment strategies have had variable results, but intramuscular promethazine has been the most effective to date with a 90% initial response rate and important reduction in residual symptoms the next flight day. Oral prophylactic treatment of crewmembers with difficulty on prior flights has had mixed results. In order to accommodate more aggressive pharmacologic management, crew medical officers receive additional training in parenteral administration of medications. Preflight medication testing is accomplished to reduce the risk of unexpected performance decrements or idiosyncratic reactions. When possible, treatment is offered in the presleep period to mask potential treatment-related drowsiness. Another phenomenon noted by crewmembers and physicians as flights have lengthened is readaptation difficulty or motion sickness on return to Earth. These problems have included nausea, vomiting, and difficulty with locomotion or coordination upon early exposure to gravity. Since landing and egress are principal concerns during this portion of the flight, these deficits are of operational concern. Postflight therapy has been directed at nausea and vomiting, and meclizine and promethazine are the principal agents used. There has been no official attempt at prophylactic treatment prior to entry. Since there is considerable individual variation in postflight deficit and since adaptation from prior flights seems to persist, it has been recommended that commanders with prior shuttle landing experience be named to flights of extended duration. PMID- 9416593 TI - Neural control of the cardiovascular system during exercise. An integrative role for the vestibular system. AB - Precise regulatory signals are required in order to adjust the cardiovascular and respiratory systems to meet the demands of exercise. Two neural mechanisms, central command and a reflex originating in contracting muscles, are known to play a large role in exercise-associated adjustments in cardiovascular and respiratory activity. The extent to which other regulatory reflexes, such as vestibulo-autonomic reflexes, are able to impact upon the cardiovascular and respiratory systems during exercise is largely unknown. Further, brain regions that may integrate these control mechanisms are only starting to be investigated. We propose that medullary brain nuclei may integrate both exercise and vestibular signals to produce a more coordinated, and therefore efficient, means of adaptation to exercise in a gravitational environment. PMID- 9416594 TI - Sleep and vestibular adaptation: implications for function in microgravity. AB - Optimal human performance depends upon integrated sensorimotor and cognitive functions, both of which are known to be exquisitely sensitive to loss of sleep. Under the microgravity conditions of space flight, adaptation of both sensorimotor (especially vestibular) and cognitive functions (especially orientation) must occur quickly--and be maintained--despite any concurrent disruptions of sleep that may be caused by microgravity itself, or by the uncomfortable sleeping conditions of the spacecraft. It is the three-way interaction between sleep quality, general work efficiency, and sensorimotor integration that is the subject of this paper and the focus of new work in our laboratory. To record sleep under field conditions including microgravity, we utilize a novel system called the Nightcap that we have developed and extensively tested on normal and sleep-disordered subjects. To perturb the vestibular system in ground-based studies, we utilize a variety of experimental conditions including optokinetic stimulation and both minifying and reversing goggle paradigms that have been extensively studied in relation to plasticity of the vestibulo-ocular reflex. Using these techniques we will test the hypothesis that vestibular adaptation both provokes and is enhanced by REM sleep under both ground-based and space conditions. In this paper we describe preliminary results of some of our studies. PMID- 9416595 TI - Human heart rate variability relation is unchanged during motion sickness. AB - In a study of 18 human subjects, we applied a new technique, estimation of the transfer function between instantaneous lung volume (ILV) and instantaneous heart rate (HR), to assess autonomic activity during motion sickness. Two control recordings of ILV and electrocardiogram (ECG) were made prior to the development of motion sickness. During the first, subjects were seated motionless, and during the second they were seated rotating sinusoidally about an earth vertical axis. Subjects then wore prism goggles that reverse the left-right visual field and performed manual tasks until they developed moderate motion sickness. Finally, ILV and ECG were recorded while subjects maintained a relatively constant level of sickness by intermittent eye closure during rotation with the goggles. Based on analyses of ILV to HR transfer functions from the three conditions, we were unable to demonstrate a change in autonomic control of heart rate due to rotation alone or due to motion sickness. These findings do not support the notion that moderate motion sickness is manifested as a generalized autonomic response. PMID- 9416596 TI - Horizontal linear and angular responses of neurons in the medial vestibular nucleus of the decerebrate cat. AB - Responses to linear accelerations in the earth-horizontal plane (typically provoked by tilts of the head or body) are characterized by a stimulus direction that produces the maximal excitation. Although changes in cardiovascular, sympathetic, and respiratory outflow are maximized during pitch, no collection of central vestibular neurons had been identified where pitch responses predominate. In the present study, response properties of neurons in the medial vestibular nucleus were examined in decerebrate cats placed on a turntable. Activation of otolith afferents was provided by constant velocity rotation with the turntable axis tilted 5 degrees from the vertical. Responsive neurons exhibited a sinusoidal modulation in their firing rate; the optimal excitatory stimulus direction was derived from responses to clockwise and counterclockwise rotations. Many of these neurons were also tested for input from horizontal semicircular canals using 0.5 Hz sinusoidal rotation about an earth-vertical axis. Of 22 tilt sensitive neurons in the medial vestibular nucleus whose optimal stimulus direction was determined, 9 were best stimulated by pitch, 10 by stimuli in one of the two vertical semicircular canal planes, and 3 by roll. Of the 33 neurons in this nucleus tested for possible convergent inputs from the otolith organs and the horizontal semicircular canals, 8 responded to both the constant velocity (otolith) stimulus and to the sinusoidal rotation, 7 appeared to receive otolith, but not horizontal canal, input, while 18 had a canal, but no otolith, response. Thus, besides serving as a relay for horizontal canal signals, the medial vestibular nucleus may also be an important relay for information about orientation within the sagittal (pitch) plane. PMID- 9416597 TI - Vestibular bibliography. PMID- 9416598 TI - Knowledge-based model of a glucosyltransferase from the oral bacterial group of mutans streptococci. AB - Mutans streptococci glucosyltransferases catalyze glucosyl transfer from sucrose to a glucan chain. We previously identified an aspartyl residue that participates in stabilizing the glucosyl transition state. The sequence surrounding the aspartate was found to have substantial sequence similarity with members of alpha amylase family. Because little is known of the protein structure beyond the amino acid sequence, we used a knowledge-based interactive algorithm, MACAW, which provided significant level of homology with alpha-amylases and glucosyltransferase from Streptococcus downei gtfI (GTF). The significance of GTF similarity is underlined by GTF/alpha-amylase residues conserved in all but one alpha-amylase invariant residues. Site-directed mutagenesis of the three GTF catalytic residues are homologous with the alpha-amylase catalytic triad. The glucosyltransferases are members of the 4/7-superfamily that have a (beta/alpha)8 barrel structure and belong to family 13 of the glycohydralases. PMID- 9416599 TI - Identification of the binding site on S100B protein for the actin capping protein CapZ. AB - The calcium-binding protein S100B binds to several potential target proteins, but there is no detailed information showing the location of the binding site for any target protein on S100B. We have made backbone assignments of the calcium-bound form of S100B and used chemical-shift changes in spectra of 15N-labeled protein to locate the site that binds a peptide corresponding to residues 265-276 from CapZ alpha, the actin capping protein. The largest chemical-shift changes are observed for resonances arising from residues around the C terminus of the C terminal helix of S100B and residues Val-8 to Asp-12 of the N-terminal helix. These residues are close to but not identical to residues that have been identified by mutational analysis to be important in other S100 protein-protein interactions. They make up a patch across the S100B dimer interface and include some residues that are quite buried in the structure of calcium-free S100B. We believe we may have identified a binding site that could be common to many S100 protein-protein interactions. PMID- 9416600 TI - Crystal structures of a marginally active thymidylate synthase mutant, Arg 126- >Glu. AB - Thymidylate synthase (TS) is a long-standing target for anticancer drugs and is of interest for its rich mechanistic features. The enzyme catalyzes the conversion of dUMP to dTMP using the co-enzyme methylenetetrahydrofolate, and is perhaps the best studied of enzymes that catalyze carbon-carbon bond formation. Arg 126 is found in all TSs but forms only 1 of 13 hydrogen bonds to dUMP during catalysis, and just one of seven to the phosphate group alone. Despite this, when Arg 126 of TS from Escherichia coli was changed to glutamate (R126E), the resulting protein had kcat reduced 2000-fold and Km reduced 600-fold. The crystal structure of R126E was determined under two conditions--in the absence of bound ligand (2.4 A resolution), and with dUMP and the antifolate CB3717 (2.2 A resolution). The first crystals, which did not contain dUMP despite its presence in the crystallization drop, displayed Glu 126 in a position to sterically and electrostatically interfere with binding of the dUMP phosphate. The second crystals contained both dUMP and CB3717 in the active site, but Glu 126 formed three hydrogen bonds to nearby residues (two through water) and was in a position that partially overlapped with the normal phosphate binding site, resulting in a approximately 1 A shift in the phosphate group. Interestingly, the protein displayed the typical ligand-induced conformational change, and the covalent bond to Cys 146 was present in one of the protein's two active sites. PMID- 9416601 TI - De novo heme proteins from designed combinatorial libraries. AB - We previously reported the design of a library of de novo amino acid sequences targeted to fold into four-helix bundles. The design of these sequences was based on a "binary code" strategy, in which the patterning of polar and nonpolar amino acids is specified explicitly, but the exact identities of the side chains is varied extensively (Kamtekar S, Schiffer JM, Xiong H, Babik JM, Hecht MH, 1993, Science 262:1680-1685). Because of this variability, the resulting collection of amino acid sequences may include de novo proteins capable of binding biologically important cofactors. To probe for such binding, the de novo sequences were screened for their ability to bind the heme cofactor. Among an initial collection of 30 binary code sequences, 15 are shown to bind heme and form bright red complexes. Characterization of several of these de novo heme proteins demonstrated that their absorption spectra and resonance Raman spectra resemble those of natural cytochromes. Because the design of these sequences is based on global features of polar/ nonpolar patterning, the finding that half of them bind heme highlights the power of the binary code strategy, and demonstrates that isolating de novo heme proteins does not require explicit design of the cofactor binding site. Because bound heme plays a key role in the functions of many natural proteins, these results suggest that binary code sequences may serve as initial prototypes for the development of large collections of functionally active de novo proteins. PMID- 9416602 TI - A flavodoxin that is required for enzyme activation: the structure of oxidized flavodoxin from Escherichia coli at 1.8 A resolution. AB - In Escherichia coli, flavodoxin is the physiological electron donor for the reductive activation of the enzymes pyruvate formate-lyase, anaerobic ribonucleotide reductase, and B12-dependent methionine synthase. As a basis for studies of the interactions of flavodoxin with methionine synthase, crystal structures of orthorhombic and trigonal forms of oxidized recombinant flavodoxin from E. coli have been determined. The orthorhombic form (space group P2(1)2(1)2(1), a = 126.4, b = 41.10, c = 69.15 A, with two molecules per asymmetric unit) was solved initially by molecular replacement at a resolution of 3.0 A, using coordinates from the structure of the flavodoxin from Synechococcus PCC 7942 (Anacystis nidulans). Data extending to 1.8-A resolution were collected at 140 K and the structure was refined to an Rwork of 0.196 and an Rfree of 0.250 for reflections with I > 0. The final model contains 3,224 non-hydrogen atoms per asymmetric unit, including 62 flavin mononucleotide (FMN) atoms, 354 water molecules, four calcium ions, four sodium ions, two chloride ions, and two Bis Tris buffer molecules. The structure of the protein in the trigonal form (space group P312, a = 78.83, c = 52.07 A) was solved by molecular replacement using the coordinates from the orthorhombic structure, and was refined with all data from 10.0 to 2.6 A (R = 0.191; Rfree = 0.249). The sequence Tyr 58-Tyr 59, in a bend near the FMN, has so far been found only in the flavodoxins from E. coli and Haemophilus influenzae, and may be important in interactions of flavodoxin with its partners in activation reactions. The tyrosine residues in this bend are influenced by intermolecular contacts and adopt different orientations in the two crystal forms. Structural comparisons with flavodoxins from Synechococcus PCC 7942 and Anaebaena PCC 7120 suggest other residues that may also be critical for recognition by methionine synthase. PMID- 9416603 TI - Linkers of secondary structures in proteins. AB - Linkers that connect repeating secondary structures fall into conformational classes based on distance and main-chain torsion clustering. A data set of 300 unique protein chains with low pairwise sequence identity was clustered into only a few groups representing the preferred motifs. The linkers of two to eight residues for the nonredundant data set are designated H-Ln-H, H-Ln-E, E-Ln-H, E Ln-E, where n is the length, H stands for alpha-helices, and E for beta-strands. Most of the clusters identified here corroborate earlier findings. However, 19 new clusters are identified in this paper, with many of them having seven and eight residue linkers. In our first analysis, the secondary structures flanking the linkers are both interacting and noninteracting and there is no precise angle of orientation between them. A second analysis was performed on a set of proteins with restricted orientations for the flanking elements, namely, mainly alpha class of proteins with orthogonal architecture. Two definite clusters are identified, one corresponding to linkers of orthogonal helices and the other to linkers of antiparallel helices. Loops forming binding sites or involved in catalytic activity are important determinants of the function of proteins. Although the structural conservation of the residues around the catalytic triad of serine proteases has been studied widely, there has not been a systematic analysis of the conformation of the loops that contain them. Residues of the catalytic triad reside in the linkers of beta-strands, with varying lengths of more than eight residues. Here, we analyze the structural conservation of such linkers by superposition, and observe a conserved structural feature of the linkers incorporating each of the three residues of the catalytic triad. PMID- 9416604 TI - Cross-strand side-chain interactions versus turn conformation in beta-hairpins. AB - A series of designed peptides has been analyzed by 1H-NMR spectroscopy in order to investigate the influence of cross-strand side-chain interactions in beta hairpin formation. The peptides differ in the N-terminal residues of a previously designed linear decapeptide that folds in aqueous solution into two interconverting beta-hairpin conformations, one with a type I turn (beta-hairpin 4:4) and the other with a type I + G1 beta-bulge turn (beta-hairpin 3:5). Analysis of the conformational behavior of the peptides studied here demonstrates three favorable and two unfavorable cross-strand side-chain interactions for beta hairpin formation. These results are in agreement with statistical data on side chain interactions in protein beta-sheets. All the peptides in this study form significant populations of the beta-hairpin 3:5, but only some of them also adopt the beta-hairpin 4:4. The formation of beta-hairpin 4:4 requires the presence of at least two favorable cross-strand interactions, whereas beta-hairpin 3:5 seems to be less susceptible to side-chain interactions. A protein database analysis of beta-hairpins 3:5 and beta-hairpins 4:4 indicates that the former occur more frequently than the latter. In both peptides and proteins, beta-hairpins 3:5 have a larger right-handed twist than beta-hairpins 4:4, so that a factor contributing to the higher stability of beta-hairpin 3:5 relative to beta-hairpin 4:4 is due to an appropriate backbone conformation of the type I + G1 beta-bulge turn toward the right-handed twist usually observed in protein beta-sheets. In contrast, as suggested previously, backbone geometry of the type I turn is not adequate for the right-handed twist. Because analysis of buried hydrophobic surface areas on protein beta-hairpins reveals that beta-hairpins 3:5 bury more hydrophobic surface area than beta-hairpins 4:4, we suggest that the right-handed twist observed in beta-hairpin 3:5 allows a better packing of side chains and that this may also contribute to its higher intrinsic stability. PMID- 9416605 TI - Analysis of temperature factor distribution in high-resolution protein structures. AB - The temperature factors obtained from X-ray refinement of proteins at high resolution show large variations from one structure to another. However, the B values expressed in units of standard deviation about their mean value (B' factor) at the C alpha atoms show remarkably characteristic frequency distribution. In all of the 110 proteins examined in this study, the frequency distribution exhibited a bimodal distribution. The peaks in the B'-factor frequency distribution occur at -1.1 and 0.4 for a bin size of 0.5. The peak at lower temperature factor corresponds largely to buried residues, whereas the peak at larger value corresponds to exposed residues. The distribution could be accurately described as a superposition of two Gaussian functions. The parameters describing the distribution are therefore characteristic of protein structures. The frequency distribution for a given amino acid over all the proteins also shows a similar bimodal distribution, although the areas under the two Gaussians differ from one amino acid to another. The area under the frequency distribution curve for any interval in B'-factor represents the propensity of the amino acid to occur in that interval. This propensity is related both to the hydrophilicity/hydrophobicity of the residue and the tendency of the residue to impose a different degree of rigidity on the polypeptide chain. The frequency distribution of stretches of high B'-factors departs appreciably from that expected for a random distribution. The correlation in the B-values of sequentially proximal residues is probably responsible for the bimodal distribution. PMID- 9416606 TI - Structural characterization of human hemoglobin crosslinked by bis(3,5 dibromosalicyl) fumarate using mass spectrometric techniques. AB - Diaspirin crosslinked hemoglobin (DCLHb) was analyzed by mass spectrometric-based techniques to identify the protein modifications effected by the crosslinking reaction with bis(3,5-dibromosalicyl) fumarate. DCLHb consists of two principal components. These components were isolated by size-exclusion chromatography and identified by measurement of their molecular weight using electrospray mass spectrometry and subsequent peptide mass mapping and mass spectrometric sequence analysis of their individual digests. Three major RP-HPLC fractions were observed from the major hemoglobin in DCLHb. Their MWs matched the MW of heme, intact hemoglobin beta-chain, and two hemoglobin alpha-chains crosslinked by a fumarate moiety, respectively. The minor HPLC peaks of DCLHb were also separated, and characterized by mass spectrometric methods. These minor components revealed additional details of the structural nature of covalent modification of DCLHb. PMID- 9416607 TI - Lack of coupling between secondary structure formation and collapse in a model polypeptide that mimics early folding intermediates, the F2 fragment of the Escherichia coli tryptophan-synthase beta chain. AB - The isolated, 101-residue long C-terminal (so called F2) fragment of the beta chain from Escherichia coli tryptophan synthase was shown previously to fold into an ensemble of conformations that are condensed, to contain large amounts of highly dynamic secondary structures, and to behave as a good model of structured intermediates that form at the very early stages of protein folding. Here, solvent perturbations were used to investigate the forces that are involved in stabilizing the secondary structure (monitored by far-UV CD) and the condensation of the polypeptide chain (monitored by dynamic light scattering) in isolated F2. It was observed that neither the ionic strength, nor the pH (between 7 and 10), nor salts of the Hofmeister series affected the global secondary structure contents of F2, whereas some of these salts affected the collapse slightly. Addition of trifluoroethanol resulted in a large increase in both the amount of secondary structure and the Stokes radius of F2. Conversely, F2 became more condensed upon raising the temperature from 4 to 60 degrees C, whereas in this temperature range, the secondary structure undergoes significant melting. These observations lead to the conclusion that, in isolated F2, there is no coupling between the hydrophobic collapse and the secondary structure. This finding will be discussed in terms of early events in protein folding. PMID- 9416608 TI - Dynamics and unfolding pathways of a hyperthermophilic and a mesophilic rubredoxin. AB - Molecular dynamics simulations in solution are performed for a rubredoxin from the hyperthermophilic archaeon Pyrococcus furiosus (RdPf) and one from the mesophilic organism Desulfovibrio vulgaris (RdDv). The two proteins are simulated at four temperatures: 300 K, 373 K, 473 K (two sets), and 500 K; the various simulations extended from 200 ps to 1,020 ps. At room temperature, the two proteins are stable, remain close to the crystal structure, and exhibit similar dynamic behavior; the RMS residue fluctuations are slightly smaller in the hyperthermophilic protein. An analysis of the average energy contributions in the two proteins is made; the results suggest that the intraprotein energy stabilizes RdPf relative to RdDv. At 373 K, the mesophilic protein unfolds rapidly (it begins to unfold at 300 ps), whereas the hyperthermophilic does not unfold over the simulation of 600 ps. This is in accord with the expected stability of the two proteins. At 473 K, where both proteins are expected to be unstable, unfolding behavior is observed within 200 ps and the mesophilic protein unfolds faster than the hyperthermophilic one. At 500 K, both proteins unfold; the hyperthermophilic protein does so faster than the mesophilic protein. The unfolding behavior for the two proteins is found to be very similar. Although the exact order of events differs from one trajectory to another, both proteins unfold first by opening of the loop region to expose the hydrophobic core. This is followed by unzipping of the beta-sheet. The results obtained in the simulation are discussed in terms of the factors involved in flexibility and thermostability. PMID- 9416609 TI - Simulating the minimum core for hydrophobic collapse in globular proteins. AB - To investigate the nature of hydrophobic collapse considered to be the driving force in protein folding, we have simulated aqueous solutions of two model hydrophobic solutes, methane and isobutylene. Using a novel methodology for determining contacts, we can precisely follow hydrophobic aggregation as it proceeds through three stages: dispersed, transition, and collapsed. Theoretical modeling of the cluster formation observed by simulation indicates that this aggregation is cooperative and that the simulations favor the formation of a single cluster midway through the transition stage. This defines a minimum solute hydrophobic core volume. We compare this with protein hydrophobic core volumes determined from solved crystal structures. Our analysis shows that the solute core volume roughly estimates the minimum core size required for independent hydrophobic stabilization of a protein and defines a limiting concentration of nonpolar residues that can cause hydrophobic collapse. These results suggest that the physical forces driving aggregation of hydrophobic molecules in water is indeed responsible for protein folding. PMID- 9416610 TI - Thermal unfolding of dodecameric glutamine synthetase: inhibition of aggregation by urea. AB - Thermal unfolding of dodecameric manganese glutamine synthetase (622,000 M(r)) at pH 7 and approximately 0.02 ionic strength occurs in two observable steps: a small reversible transition (Tm approximately 42 degrees C; delta H approximately equal to 0.9 J/g) followed by a large irreversible transition (Tm approximately 81 degrees C; delta H approximately equal to 23.4 J/g) in which secondary structure is lost and soluble aggregates form. Secondary structure, hydrophobicity, and oligomeric structure of the equilibrium intermediate are the same as for the native protein, whereas some aromatic residues are more exposed. Urea (3 M) destabilizes the dodecamer (with a tertiary structure similar to that without urea at 55 degrees C) and inhibits aggregation accompanying unfolding at < or = 0.2 mg protein/mL. With increasing temperature (30-70 degrees C) or incubation times at 25 degrees C (5-35 h) in 3 M urea, only dodecamer and unfolded monomer are detected. In addition, the loss in enzyme secondary structure is pseudo-first-order (t1/2 = 1,030 s at 20.0 degrees C in 4.5 M urea). Differential scanning calorimetry of the enzyme in 3 M urea shows one endotherm (Tmax approximately 64 degrees C; delta H = 17 +/- 2 J/g). The enthalpy change for dissociation and unfolding agrees with that determined by urea titrations by isothermal calorimetry (delta H = 57 +/- 15 J/g; Zolkiewski M, Nosworthy NJ, Ginsburg A, 1995, Protein Sci 4: 1544-1552), after correcting for the binding of urea to protein sites exposed during unfolding (-42 J/g). Refolding and assembly to active enzyme occurs upon dilution of urea after thermal unfolding. PMID- 9416611 TI - Demonstration of protein-protein interaction specificity by NMR chemical shift mapping. AB - Chemical shift mapping is becoming a popular method for studying protein-protein interactions in solution. The technique is used to identify putative sites of interaction on a protein surface by detecting chemical shift perturbations in simple (1H, 15N)-HSQC NMR spectra of a uniformly labeled protein as a function of added (unlabeled) target protein. The high concentrations required for these experiments raise questions concerning the possibility for non-specific interactions being detected, thereby compromising the information obtained. We demonstrate here that the simple chemical shift mapping approach faithfully reproduces the known functional specificities among pairs of closely related proteins from the phosphoenolpyruvate:sugar phosphotransferase systems of Escherichia coli and Bacillus subtilis. PMID- 9416612 TI - OLDERADO: on-line database of ensemble representatives and domains. On Line Database of Ensemble Representatives And DOmains. AB - In cases where the structure of a single protein is represented by an ensemble of conformations, there is often a need to determine the common features and to choose a "representative" conformation. This occurs, for example, with structures determined by NMR spectroscopy, analysis of the trajectory from a molecular dynamics simulation, or an ensemble of structures produced by comparative modeling. We reported previously automatic methods for (1) defining the atoms with low spatial variance across an ensemble (i.e., the "core" atoms) and the domains in which these atoms lie, and (2) clustering an ensemble into conformationally related subfamilies. To extend the utility of these methods, we have developed a freely available server on the World Wide Web at http:/(/)neon.chem.le.ac.uk/olderado/. This (1) contains an automatically generated database of representative structures, core atoms, and domains determined for 449 ensembles of NMR-derived protein structures in the Protein Data Bank (PDB) in May 1997, and (2) allows the user to upload a PDB-formatted file containing the coordinates of an ensemble of structures. The server returns in real time: (1) information on the residues constituting domains: (2) the structures that constitute each conformational subfamily; and (3) an interactive java-based three-dimensional viewer to visualise the domains and clusters. Such information is useful, for example, when selecting conformations to be used in comparative modeling and when choosing parts of structures to be used in molecular replacement. Here we describe the OLDERADO server. PMID- 9416613 TI - alpha-Hemolysin, gamma-hemolysin, and leukocidin from Staphylococcus aureus: distant in sequence but similar in structure. AB - alpha-Hemolysin from Staphylococcus aureus assembles from a water-soluble, monomeric species to a membrane-bound heptamer on the surface of target cells, creating water-filled channels that lead to cell death and lysis. Staphylococcus aureus also produces the gamma-hemolysin and leukocidin toxins, which function as two component toxins in the disruption and lysis of erythrocytes and leukocytes. Analysis of the aligned sequences of alpha-hemolysin, gamma-hemolysin, and leukocidin in the context of the alpha-hemolysin heptamer structure supports the conclusion that even though the level of sequence identity between alpha hemolysin and the gamma-hemolysin and leukocidin toxins is in the so-called twilight zone, the three-dimensional structures of the protomers are probably conserved. By analogy with alpha-hemolysin, gamma-hemolysin and leukocidin may also form oligomeric, transmembrane channels in which an antiparallel beta-barrel constitutes the primary membrane-embedded domain. PMID- 9416614 TI - Crystallization and preliminary X-ray diffraction analysis of arginyl-tRNA synthetase from Escherichia coli. AB - Arginyl-tRNA Synthetase, a class I aminoacyl tRNA synthetase playing a crucial role in protein biosynthesis, has been crystallized for the first time. Polyethylene glycol (PEG) was used as a precipitant, and the crystallization proceeded at pH 6.5. These single crystals diffracted to 2.8 A with a rotating anode X-ray source and R-axis IIc image plate detector. They have an orthorhombic space group P2(1)2(1)2 with unit cell parameters of a = 251.51 A, b = 53.12 A, and c = 52.35 A. A complete native data set has been collected at 3.1 A resolution for these crystals. PMID- 9416615 TI - A diverse superfamily of enzymes with ATP-dependent carboxylate-amine/thiol ligase activity. AB - The recently developed PSI-BLAST method for sequence database search and methods for motif analysis were used to define and expand a superfamily of enzymes with an unusual nucleotide-binding fold, referred to as palmate, or ATP-grasp fold. In addition to D-alanine-D-alanine ligase, glutathione synthetase, biotin carboxylase, and carbamoyl phosphate synthetase, enzymes with known three dimensional structures, the ATP-grasp domain is predicted in the ribosomal protein S6 modification enzyme (RimK), urea amidolyase, tubulin-tyrosine ligase, and three enzymes of purine biosynthesis. All these enzymes possess ATP-dependent carboxylate-amine ligase activity, and their catalytic mechanisms are likely to include acylphosphate intermediates. The ATP-grasp superfamily also includes succinate-CoA ligase (both ADP-forming and GDP-forming variants), malate-CoA ligase, and ATP-citrate lyase, enzymes with a carboxylate-thiol ligase activity, and several uncharacterized proteins. These findings significantly extend the variety of the substrates of ATP-grasp enzymes and the range of biochemical pathways in which they are involved, and demonstrate the complementarity between structural comparison and powerful methods for sequence analysis. PMID- 9416616 TI - Crystal structure of heat-labile enterotoxin from Escherichia coli with increased thermostability introduced by an engineered disulfide bond in the A subunit. AB - Cholera toxin (CT) produced by Vibrio cholerae and heat-labile enterotoxin (LT I), produced by enterotoxigenic Escherichia coli, are AB5 heterohexamers with an ADP-ribosylating A subunit and a GM1 receptor binding B pentamer. These toxins are among the most potent mucosal adjuvants known and, hence, are of interest both for the development of anti-diarrheal vaccines against cholera or enterotoxigenic Escherichia coli diarrhea and also for vaccines in general. However, the A subunits of CT and LT-I are known to be relatively temperature sensitive. To improve the thermostability of LT-I an additional disulfide bond was introduced in the A1 subunit by means of the double mutation N40C and G166C. The crystal structure of this double mutant of LT-I has been determined to 2.0 A resolution. The protein structure of the N40C/G166C double mutant is very similar to the native structure except for a few local shifts near the new disulfide bond. The introduction of this additional disulfide bond increases the thermal stability of the A subunit of LT-I by 6 degrees C. The enhancement in thermostability could make this disulfide bond variant of LT-I of considerable interest for the design of enterotoxin-based vaccines. PMID- 9416617 TI - Crystal structure of a non-toxic mutant of heat-labile enterotoxin, which is a potent mucosal adjuvant. AB - Two closely related bacterial toxins, heat-labile enterotoxin (LT-I) and cholera toxin (CT), not only invoke a toxic activity that affects many victims worldwide but also contain a beneficial mucosal adjuvant activity that significantly enhances the potency of vaccines in general. For the purpose of vaccine design it is most interesting that the undesirable toxic activity of these toxins can be eliminated by the single-site mutation Ser63Lys in the A subunit while the mucosal adjuvant activity is still present. The crystal structure of the Ser63Lys mutant of LT-I is determined at 2.0 A resolution. Its structure appears to be essentially the same as the wild-type LT-I structure. The substitution Ser63Lys was designed, based on the wild-type LT-I crystal structure, to decrease toxicity by interfering with NAD binding and/or catalysis. In the mutant crystal structure, the newly introduced lysine side chain is indeed positioned such that it could potentially obstruct the productive binding mode of the substrate NAD while at the same time its positive charge could possibly interfere with the critical function of nearby charged groups in the active site of LT-I. The fact that the Ser63Lys mutant of LT-I does not disrupt the wild-type LT-I structure makes the non-toxic mutant potentially suitable, from a structural point of view, to be used as a vaccine to prevent enterotoxigenic E. coli infections. The structural similarity of mutant and wild-type toxin might also be the reason why the inactive Ser63Lys variant retains its adjuvant activity. PMID- 9416618 TI - Expression, characterization, and crystallization of a member of the novel phospholipase D family of phosphodiesterases. AB - A family of phospholipase D (PLD) proteins has recently been identified (Koonin, 1996; Ponting & Kerr, 1996) based upon amino acid sequence identity. This family includes human and plant PLDs, proteins encoded by open reading frames in pathogenic viruses and bacteria, as well as an endonuclease. The endonuclease, known as Nuc, is encoded by the IncN plasmid, pKM101, present in Salmonella typhimurium. The recombinant Nuc protein has been expressed and purified from Escherichia coli. The amino-terminal sequencing of the purified protein indicated that the mature protein started from the 23rd residue of the predicted sequence, suggesting that the protein is proteolytically processed during export to the periplasmic space. The recombinant enzyme was able to hydrolyze both double and single-strand DNA and an artificial substrate, bis(4-nitrophenyl) phosphate, which contains a phosphodiester bond. The enzyme activity was not inhibited in the presence of EDTA and was not regulated by divalent cations. The purified protein has been crystallized by hanging drop vapor diffusion methods, and those crystals diffract to 1.9 A resolution. PMID- 9416619 TI - Crystallization of acetate kinase from Methanosarcina thermophila and prediction of its fold. AB - The unique biochemical properties of acetate kinase present a classic conundrum in the study of the mechanism of enzyme-catalyzed phosphoryl transfer. Large, single crystals of acetate kinase from Methanosarcina thermophila were grown from a solution of ammonium sulfate in the presence of ATP. The crystals diffract to beyond 1.7 A resolution. Analysis of X-ray data from the crystals is consistent with a space group of C2 and unit cell dimensions a = 181 A, b = 67 A, c = 83 A, beta = 103 degrees. Diffraction data have been collected from the crystals at 110 and 277 K. Data collected at 277 K extend to lower resolution, but are more reproducible. The orientation of a noncrystallographic two-fold axis of symmetry has been determined. Based on an analysis of the predicted amino acid sequences of acetate kinase from several organisms, we hypothesize that acetate kinase is a member of the sugar kinase/actin/hsp70 structural family. PMID- 9416620 TI - Crystallization of the first three domains of the human insulin-like growth factor-1 receptor. AB - The insulin-like growth factor-1 receptor (IGF-1R) is a tyrosine kinase receptor of central importance in cell proliferation. A fragment (residues 1-462) comprising the L1-cysteine rich-L2 domains of the human IGF-1R ectodomain has been overexpressed in glycosylation-deficient Lec8 cells and has been affinity purified via a c-myc tag followed by gel filtration. The fragment was recognized by two anti-IGF-1R monoclonal antibodies, 24-31 and 24-60, but showed no detectable binding of IGF-1 or IGF-2. Isocratic elution of IGF-1R/462 on anion exchange chromatography reduced sample heterogeneity, permitting the production of crystals that diffracted to 2.6 A resolution with cell dimensions a = 77.0 A, b = 99.5 A, c = 120.1 A, and space group P2(1)2(1)2(1). PMID- 9416621 TI - Complete measurement of the pKa values of the carboxyl and imidazole groups in Bacillus circulans xylanase. AB - Electrostatic interactions in proteins can be dissected experimentally by determining the pKa values of their constituent ionizable amino acids. To complement previous studies of the glutamic acid and histidine residues in Bacillus circulans xylanase (BCX), we have used NMR methods to measure the pKa s of the seven aspartic acids and the C-terminus of this protein. The pKa s of these carboxyls are all less than the corresponding values observed with random coil polypeptides, indicating that their ionization contributes favorably to the stability of the folded enzyme. In general, the aspartic acids with the most reduced pKa s are those with limited exposure to the solvent and a high degree of conservation among homologous xylanases. Most dramatically, Asp 83 and Asp 101 have pKa s < 2 and thus remain deprotonated in native BCX under all conditions examined. Asp 83 is completely buried, forming a strong salt bridge with Arg 136. In contrast, Asp 101 is located on the surface of the protein, stabilized in the deprotonated form by an extensive network of hydrogen bonds involving an internal water molecule and the neutral side-chain and main-chain atoms of Ser 100 and Thr 145. These data provide a complete experimental database for theoretical studies of the ionization behavior of BCX under acidic conditions. PMID- 9416622 TI - Crystal structures of the cadmium- and mercury-substituted metallo-beta-lactamase from Bacteroides fragilis. AB - The metallo-beta-lactamases require zinc or cadmium for hydrolyzing beta-lactam antibiotics and are inhibited by mercurial compounds. To data, there are no clinically useful inhibitors of this class of enzymes. The crystal structure of the Zn(2+)-bound enzyme from Bacteroides fragilis contains a binuclear zinc center in the active site. A hydroxide, coordinated to both zinc atoms, is proposed as the moiety that mounts the nucleophilic attack on the carbonyl carbon atom of the beta-lactam ring. To study the metal coordination further, the crystal structures of a Cd(2+)-bound enzyme and of an Hg(2+)-soaked zinc containing enzyme have been determined at 2.1 A and 2.7 A, respectively. Given the diffraction resolution, the Cd(2+)-bound enzyme exhibits the same active-site architecture as that of the Zn(2+)-bound enzyme, consistent with the fact that both forms are enzymatically active. The 10-fold reduction in activity of the Cd(2+)-bound molecule compared with the Zn(2+)-bound enzyme is attributed to fine differences in the charge distribution due to the difference in the ionic radii of the two metals. In contrast, in the Hg(2+)-bound structure, one of the zinc ions, Zn2, was ejected, and the other zinc ion, Zn1, remained in the same site as in the 2-Zn(2+)-bound structure. Instead of the ejected zinc, a mercury ion binds between Cys 104 and Cys 181, 4.8 A away from Zn1 and 3.9 A away from the site where Zn2 is located in the 2-Zn(2+)-bound molecule. The perturbed binuclear metal cluster explains the inactivation of the enzyme by mercury compounds. PMID- 9416624 TI - HIV-1 Nef protein: purification, crystallizations, and preliminary X-ray diffraction studies. AB - Human immunodeficiency virus Nef protein accelerates virulent progression of AIDS by its interaction with specific cellular proteins involved in cellular activation and signal transduction. Here we report the purification and crystallization of the conserved core of HIV-1LAI Nef protein in the unliganded form and in complex with the wild-type SH3 domain of the P59fyn protein-tyrosine kinase. One-dimensional NMR experiments show that full-length protein and truncated fragment corresponding to the product of HIV-1 protease cleavage have a well-folded compact tertiary structure. The ligand-free HIV-1 Nefcore protein forms cubic crystals belonging to space group P23 with unit cell dimensions of a = b = c = 86.4 A. The Nef-Fyn SH3 cocrystals belong to the space group P6(1)22 or its enantiomorph, P6(5)22, with unit cell dimensions of a = b = 108.2 A and c = 223.7 A. Both crystal forms diffract to a resolution limit of 3.0 A resolution using synchrotron radiation, and are thus suitable for X-ray structure determination. PMID- 9416623 TI - Crystallization and preliminary X-ray studies of I-PpoI: a nuclear, intron encoded homing endonuclease from Physarum polycephalum. AB - The homing endonuclease I-PpoI is encoded by an optional third intron, Pp LSU 3, found in nuclear, extrachromosomal copies of the Physarum polycephalum 26S rRNA gene. This endonuclease promotes the lateral transfer or "homing" of its encoding intron by recognizing and cleaving a partially symmetric, 15 bp homing site in 26S rDNA alleles that lack the Pp LSU 3 intron. The open reading frame encoding I PpoI has been subcloned, and the endonuclease has been overproduced in E. coli. Purified recombinant I-PpoI has been co-crystallized with a 21 bp homing site DNA duplex. The crystals belong to space group P3(1)21, with unit cell dimensions a = b = 114 A, c = 89 A. The results of initial X-ray diffraction experiments indicate that the asymmetric unit contains an enzyme homodimer and one duplex DNA molecule, and that the unit cell has a specific volume of 3.4 A3/dalton. These experiments also provide strong evidence that I-PpoI contains several bound zinc ions as part of its structure. PMID- 9416626 TI - Adult age differences in controlled and automatic memory processing. AB - The memory performance of groups of younger, middle-aged, and older participants was tested on indirect and direct tests of word stem completion and on a process dissociation task. As expected, on the direct tests of stem completion, older participants had lower scores than the younger and middle-aged groups. Age effects were also found on the indirect word completion test. The process dissociation task allowed memory performance to be divided into controlled and automatic processing components. Estimates of automatic processing were comparable for the three groups, but there was an age effect for controlled processing, with the middle-aged and older groups differing from the younger group. These results confirm the findings of J. M. Jennings and L. L. Jacoby (1993) and suggest that the decline in conscious processing efficiency begins in middle age. PMID- 9416625 TI - Sequential and coordinative complexity in time-accuracy functions for mental arithmetic. AB - Time-accuracy functions for tasks involving single-digit mental addition and subtraction were derived in a sample of 18 younger (mean age = 21.7 years) and 16 older adults (mean age = 68.8 years). Sequential complexity was manipulated by varying the number of operations (5 vs. 10); coordinative complexity was induced by bracketing. Age differences were apparent in the coordinative conditions, even though no age difference was present in the sequential conditions. This indicates that the age difference under conditions of high coordinative demands could not be attributed solely to a decline in basic speed of processing. The Age x Complexity interaction was due to larger onset times and lower asymptotic performance by the older adults in the coordinative conditions but not due to to rate of approach to the asymptote. This implies that coordinative demands do not differentially hurt access from semantic memory in older adults; however, coordinative demands do have disproportionately negative consequences for computation speed and self-monitoring in older adults. PMID- 9416627 TI - Loneliness and nursing home admission among rural older adults. AB - In this study, the authors tested the relation between loneliness and subsequent admission to a nursing home over a 4-year time period in a sample of approximately 3,000 rural older Iowans. Higher levels of loneliness were found to increase the likelihood of nursing home admission and to decrease the time until nursing home admission. The influence of extremely high loneliness on nursing home admission remained statistically significant after controlling for other variables, such as age, education, income, mental status, physical health, morale, and social contact, that were also predictive of nursing home admission. Several mechanisms are proposed to explain the link between extreme loneliness and nursing home admission. These include loneliness as a precipitant of declines in mental and physical health and nursing home placement as a strategy to gain social contact with others. Implications for preventative interventions are discussed. PMID- 9416628 TI - Emotion and aging: experience, expression, and control. AB - Age differences in emotional experience, expression, and control were investigated in 4 studies. A community sample of 127 African Americans and European Americans (ages 19-96 years) was used in Study 1; a community sample of 82 Chinese Americans and European Americans (ages 20-85 years) was used in Study 2; a community sample of 49 Norwegians drawn from 2 age groups (ages 20-35 years and 70+ years) was used in Study 3; and a sample of 1,080 American nuns (ages 24 101 years) was used in Study 4. Across studies, a consistent pattern of age differences emerged. Compared with younger participants, older participants reported fewer negative emotional experiences and greater emotional control. Findings regarding emotional expressivity were less consistent, but when there were age differences, older participants reported lesser expressivity. Results are interpreted in terms of increasingly competent emotion regulation across the life span. PMID- 9416629 TI - Everyday activity parameters and competence in older adults. AB - Parameters of everyday activities in relation to cognitive, social, and emotional competence were examined in 2 studies. The parameters included frequency, difficulty, importance, intentions for future activities, changes in past activities, and ability of performance. The challenge hypothesis, in which performance of optional activities experienced as moderately difficult is associated with greatest well-being, was also tested. Two samples of older adults completed a life history interview and measures of psychological functioning. Parameters of activities necessary for maintaining an independent engaged lifestyle were measured by the Everyday Activities Questionnaire. In both studies, competence variables helped explain activity parameters independently of age and demographic variables. There was no support for the challenge hypothesis in either study. PMID- 9416630 TI - Perceived problems for self and others: self-protection by social downgrading throughout adulthood. AB - In this study the authors investigated social downgrading as reflected in the difference between perceptions about the self and about "most people my age." A large cross-national probability sample of adults at different age levels throughout adulthood provided ratings of perceived problems expected for the self and for "most other people my age" with regard to 12 domains of life (e.g., health, marriage, and job). Results showed that with regard to all domains, younger, middle-aged, and older adults believed other people's problems to be more serious than their own problems in these domains. Social downgrading was particularly pronounced for those domains for which a given participant experienced problems himself or herself. This self-protection tendency under threat was particularly pronounced in the older adults. The function and adaptive values for social downgrading across adulthood and old age are discussed. PMID- 9416631 TI - Do performance strategies mediate age-related differences in associative learning? AB - Associative learning is a basic component of most learning tasks and has been shown to decline with age. The authors examined associative learning for younger and older adults by using a noun-pair task. Interim testing and prior practice on a similar task were the manipulated variables. Participants were encouraged to use an efficient retrieval strategy. Interim tests provided the motivation to learn the information, whereas prior practice on a similar task was presumed to make the task easier. The authors examined these variables both independently and interactively. For younger adults, performance benefited little from prior practice but did benefit from interim testing. For older adults, interim tests were beneficial for development of a retrieval strategy irrespective of prior training. Prior training proved beneficial for development of a retrieval strategy in the absence of interim tests. Thus, task parameters influenced the performance strategy (and learning), especially of older adults. PMID- 9416632 TI - Age differences in implicit learning of higher order dependencies in serial patterns. AB - 3 experiments examined serial pattern learning in younger and older adults. Unlike the usual repeating pattern, the sequences alternated between events from a repeating pattern and those determined randomly. The results indicated that no one was able to describe the regularity, but with practice every individual in all 3 age groups (including old old) became faster, more accurate, or both, on pattern trials than on random trials. Although this indicates that adults of all ages are able to learn second-order statistical dependencies in a sequence, age related deficits were obtained in the magnitude of pattern learning. There were also age differences in what was learned, with only younger people revealing sensitivity to higher order statistical dependencies in the sequence. In addition, whereas younger people revealed evidence of their pattern learning in a subsequent conceptually driven production test, young-old and old-old people did not. PMID- 9416633 TI - Social self-efficacy and short-term variability in social relationships: the MacArthur successful aging studies. AB - Dealing with others entails both stability and short-term variability of the functions and outcomes of social relationships. The authors argue that patterns of short-term intraindividual variability in social relationships and self efficacy beliefs contribute interpretable information about social adaptation. On the basis of 23 repeated weekly measurements of a sample of 32 participants ages 56 to 88 years, the authors examined the extent to which fluctuations in perceived relational outcomes are related to fluctuations of social self efficacy. Results showed that individuals differ systematically in respect to the extent to which they experience and display fluctuations in self-efficacy and availability of social relationships. Moreover, when individuals perceive others to be available across time, social self-efficacy beliefs are stronger and fluctuate less across time. PMID- 9416634 TI - Continued inhibitory capacity throughout adulthood: conceptual negative priming in younger and older adults. AB - Two negative priming experiments in older and younger adults are reported. Participants in Experiment 1, involving both positive and negative priming conditions, showed both types of priming. There were no significant differences between age groups. If anything, older participants showed more negative priming. In Experiment 2, involving only negative priming conditions, similar results were obtained. Our findings rule out possible effects of experimental conditions that episodic retrieval theorists have suggested might account for negative priming in older adults. Although our results may be consistent with an explanation of negative priming in older adults by an expansively specified theory of episodic retrieval, they are at least as consistent with the view that inhibitory processes are intact in older adults. In light of these findings, conflicting empirical results and alternative views of negative priming in older adults are examined. PMID- 9416635 TI - Interrelations of age, self-reported health, speed, and memory. AB - Contributions of self-reported health to adult age differences in perceptual speed and memory were assessed for 301 adults ages 20-90. Participants were asked 4 health status questions, given 3 perceptual speed tests, 2 working memory tests, and 2 memory tests. Self-reported health was found to predict speed better than it predicted memory. Covariance structural equation modeling was used to assess the relations among age, self-reported health, perceptual speed, working memory, and memory. The results support the hypothesis that any effects of self reported health on age differences in memory are mediated by perceptual speed. PMID- 9416636 TI - Everyday problem solving across the adult life span: influence of domain specificity and cognitive appraisal. AB - Differences in problem-solving strategies for situations varying in three domains, consumer, home management, and conflict with friends, were examined among younger, middle-aged, and older adults. In addition, this study examined the influence of perceived ability to resolve the problem, controllability, and causal attributions on strategy selection. In the 2 instrumental domains, older adults were more problem focused in their approach than adolescents and younger adults, whereas adolescents and younger adults selected more passive-dependent strategies. In the more interpersonal domain, conflict with friends, older adults tended to select avoidant-denial strategies more so than younger adults. Finally, across domains, the greater the perceived ability to resolve a problem the less the avoidant-denial strategy was selected. The importance of distinguishing between social and instrumental problem solving and of examining the cognitive appraisal of a problem situation are discussed. PMID- 9416637 TI - Age differences in stages of attributional processing. AB - This study introduces an attributional processing approach to study age differences in dispositional attributions. Dispositional attributions made in the context of relationship vignettes were examined among younger and older adults in 2 conditions (immediate and delayed rating conditions). By using a direct assessment of a 2-step process for making dispositional attributions, it was inferred that a spontaneous adjustment stage occurred following an initial characterization stage as a function of age group and content of vignettes. Older adults made lower dispositional ratings if they were given more time to think about the situations than if asked to make an immediate judgment. By contrast, younger adults made higher dispositional ratings if they were given more time to make the judgments. Qualitative analyses of schemas elicited by a subsample of participants for each vignette suggested a relationship between dispositional attributional ratings and content-evoked schemas. PMID- 9416638 TI - Age differences in visual search for feature, conjunction, and triple-conjunction targets. AB - The authors examined the ability of younger and older adults to search for targets defined by single features (feature search), conjunctions of 2 features (conjunction search), and conjunctions of 3 features (triple-conjunction search). Feature search was relatively age-invariant, with both older and younger adults displaying shallow search slopes. However, older adults did show reduced search rates for many conjunction targets. Interestingly, both older and younger adults benefited equivalently when an extra feature was available to define the conjunction target. That is, the relative amount of improvement in search performance was similar for younger and older adults when the triple-conjunction search was compared to the conjunction search. These results are discussed in terms of age-related differences in the effectiveness of bottom-up and top-down mechanisms that support visual search. PMID- 9416639 TI - Differential item functioning in the Mini-Mental State Examination in English- and Spanish-speaking older adults. AB - The purpose of this study was to determine if the Mini-Mental State Examination (MMS; M. F. Folstein, S. E. Folstein, & P. R. McHugh, 1975) demonstrates item bias with respect to measuring cognitive functioning of older Hispanics and non Hispanics. Assessment of differential item functioning (DIF) of individual MMS items across 3 language/ethnicity groups (English test administration/non Hispanic ethnicity, English test administration/Hispanic ethnicity, and Spanish test administration/Hispanic ethnicity) was performed by using a logistic regression procedure. Fifteen of the 26 MMS items were significantly related to total score and were shown to provide unbiased measurement across the 3 groups. Normative data are presented for older Hispanics (n = 365) and non-Hispanics (n = 388) on the raw MMS, a 15-item version in which items with significant DIF were eliminated, and a total score statistically adjusted for effects of education and age. PMID- 9416640 TI - Schizophrenia, antipsychotics, and aging. PMID- 9416641 TI - Regional cerebral blood flow in late-onset schizophrenia: a SPECT study using 99mTc-HMPAO. AB - Functional neuroimaging studies have been performed in many young patients with schizophrenia, but late-onset schizophrenia (LOS) remains largely unexamined by these techniques. We predicted that LOS would demonstrate regional cerebral blood flow (rCBF) abnormalities similar to those seen in early-onset schizophrenia (EOS), but with a basis in demonstrable coarse brain disease. The subjects were 15 LOS and 7 EOS patients and 27 healthy controls. Each was given a detailed clinical and neuropsychological assessment and underwent MRI and Tc99m-HMPAO single photon emission computed tomography (SPECT) scans. The LOS subjects had a significantly lower cerebral hemispheric perfusion than controls, with a lower perfusion in the frontal and temporal lobes bilaterally. The LOS group also had significantly lower left-to-right hemisphere blood flow ratios. EOS subjects had a lower frontal perfusion than the controls, which was significant in the left frontal region. The temporal perfusion in the EOS subjects was greater than in the LOS group, and not different from the control subjects. Left temporal perfusion was the most discriminating variable between LOS and control subjects on logistic regression. Correlations of perfusion with MRI were generally low with the exception that the asymmetry indices were significantly correlated, and basal ganglia perfusion correlated with basal ganglia hyperintensities on MRI. The total cerebral perfusion index correlated significantly with the mini-mental state examination (MMSE) score, and the temporal lobe perfusion correlated with MMSE scores and some verbal memory measures. In the schizophrenic groups, perfusion correlated nonsignificantly with symptom profiles. We conclude that our findings of temporal and frontal rCBF abnormalities, especially on the left side, in LOS are similar to those reported in schizophrenia in general. The results do not provide evidence for coarse brain disease underlying the rCBF abnormalities in LOS, or support the specificity of these abnormalities for particular subsyndromes of schizophrenia. PMID- 9416642 TI - Effects of schizophrenia and aging on pupillographic measures of working memory. AB - The amount of cognitive resources used to perform a task can be indexed as changes in pupil size. In a previous study, we examined pupillary response measures of slave store and central executive cognitive resources during a working memory task and found abnormally reduced utilization of these resources in schizophrenia. In the present study, multiple regression analyses were performed to examine the independent and combined effects of aging and schizophrenia on pupillary response and recall measures in a larger sample of community-dwelling schizophrenia patients. Schizophrenia was associated with a significant decline in working memory capacity, and an additional moderate decline was associated with aging, but these two factors did not interact. Baseline pupil size was significantly correlated with symptom severity, independent of medication. However, pupillary responses evoked by the working memory task and recall scores were not related to symptom severity. Results were consistent with an additive, rather than a synergistic, relationship between aging and schizophrenia, and suggested that working memory impairment in noninstitutionalized outpatients with schizophrenia may remain stable across symptom status and across the life span. PMID- 9416643 TI - Functional brain imaging and aging in schizophrenia. AB - Patients with schizophrenia, even early in the course of illness, show some similarities in their brain-imaging findings to those in older normal controls: for example, ventricular enlargement and diminished functional activity in the frontal cortex. These findings suggest a possible similarity between the normal aging process and schizophrenia, or the possible existence in some schizophrenic patients of progressive processes that prematurely affect the brain in ways analogous to normal aging. In contrast to other brain regions, the basal ganglia in schizophrenia may show an atypical pattern of volumetric and metabolic change over time, possibly because of the effects of neuroleptic treatment. However, age and illness duration are highly correlated in our samples. Interpretation of imaging results is limited by the lack of studies in an adequate number of either first-break or older schizophrenic patients and the dearth of studies with longitudinal designs. PMID- 9416644 TI - Review of functional magnetic resonance imaging in schizophrenia. AB - Functional magnetic resonance imaging (fMRI) holds great promise for assessing temporal changes in brain activity using various challenge paradigms. In this report, we review the 14 studies (eight of them abstracts) that comprise the fMRI literature available to date relating to schizophrenia. Twelve of the 14 investigations examined changes in blood-oxygen-level-dependent (BOLD) contrast: two examined blood volume. Eight of the 12 BOLD studies relied on lower-order cognitive processing to measure activation (involving sensory or motor areas), whereas four used higher-order tasks (word production, auditory processing, and subspan word recall involving multiple brain areas). Although the variability in tasks used, brain regions studied, imaging methods used, patient characteristics reported, and methods of reporting significance precluded a full meta-analysis, we re-analyzed these published data to compute effect sizes. In most studies, resting blood volume and BOLD changes, regardless of the complexity of the cognitive task, appeared to differ between patients with schizophrenia and control subjects. PMID- 9416645 TI - Glutamate in schizophrenia: clinical and research implications. AB - The excitatory amino acids, glutamate and aspartate, are of interest to schizophrenia research because of their roles in neurodevelopment, neurotoxicity and neurotransmission. Recent evidence suggests that densities of glutamatergic receptors and the ratios of subunits composing these receptors may be altered in schizophrenia, although it is unclear whether these changes are primary or compensatory. Agents acting at the phencyclidine binding site of the NMDA receptor produce symptoms of schizophrenia in normal subjects, and precipitate relapse in patients with schizophrenia. The improvement of negative symptoms with agents acting at the glycine modulatory site of the NMDA receptor, as well as preliminary evidence that clozapine may differ from conventional neuroleptic agents in its effects on glutamatergic systems, suggest that clinical implications may follow from this model. While geriatric patients may be at increased risk for glutamate-mediated neurotoxicity, very little is known about the specific relevance of this model to geriatric patients with schizophrenia. PMID- 9416646 TI - Cellular and molecular neuropathology of schizophrenia: new directions from developmental neurobiology. AB - Schizophrenia is in essence a developmental disorder, but an unusual one in that the onset of symptoms is markedly delayed. Neuropathologic studies of the brain in schizophrenia have revealed subtle abnormalities that may reflect abnormal neuronal development. A more detailed examination of the cellular and molecular pathology of schizophrenia has been limited by the lack of informative markers that might allow a more complete understanding of the brain defects that characterize this disorder. Recent advances in molecular biology have made available a growing number of probes for examining the expression of specific gene products in brain tissue by using in situ hybridization and immunohistochemistry using antibodies to recombinant antigens. Several recently cloned neural genes are expressed in the forebrain regions which have been implicated in schizophrenia, and may have significant roles in brain development or function. Selected neurotransmitter receptors, neurotrophins and their receptors, and transcription factors of the POU and MADS families are promising candidates for future studies of the cellular and molecular neuropathology of schizophrenia. PMID- 9416647 TI - A comparative study of elderly patients with schizophrenia and bipolar disorder in nursing homes and the community. AB - This study compared the functioning of 188 elderly schizophrenic and bipolar disorder patients living in nursing homes and the community. Residential status and diagnostic groups were compared on measures of symptomatology, cognitive impairment, functional impairment, and behavior problems. In general, the diagnostic groups differed in symptoms, while most differences in living setting were related to cognition, functioning, and behavior. Nursing home status was significantly associated with more severe overall symptom ratings, worse cognitive impairment, greater functional impairment, more aggressive behaviors, and marital status of having never married. Self-care skills, community living skills, and marital status were most uniquely predictive of nursing home residence. However, cognitive deficits were strongly predictive of both self-care and community living skills, explaining approximately half of the variance in these variables. The implications of these findings for the treatment of elderly patients with schizophrenia and other severe mental illnesses are discussed. PMID- 9416648 TI - Lateralized hemispheric dysfunction in the major psychotic disorders: historical perspectives and findings from a study of motor asymmetry in older patients. AB - Differences in functioning between the two cerebral hemispheres have been reported for more than a century. In recent decades, issues related to lateralized dysfunction have been raised in psychiatric illnesses such as schizophrenia and bipolar disorder. In particular, evidence suggests that schizophrenia may be particularly associated with left hemisphere dysfunction and bipolar disorder with right hemisphere dysfunction. We discuss these issues, along with a conceptual framework for integrating hypotheses about the relationship between the major psychotic illnesses based on a two-dimensional continuum. We also present new findings from our study of motor asymmetry in older patients with psychosis that support this framework. Our results indicate that schizophrenia may be associated with left hemisphere pathology to a greater extent than right, whereas the reverse may occur in bipolar disorder. PMID- 9416649 TI - Self-reported social functioning among older patients with schizophrenia. AB - The purpose of this study was to evaluate the utility of a self-report measure of social functioning as an outcome measure for older schizophrenia patients. Sixty five schizophrenia patients and 39 healthy controls, ranging in age from 45 to 81 years, were evaluated using a modified Social Adjustment Scale (SAS-M), Scales for Assessment of Positive and Negative Symptoms, Depression Subscale of the Brief Symptom Inventory, Mini-Mental State Examination, Dementia Rating Scale, measures of social support, and measures of background variables. Compared with controls, fewer patients with schizophrenia engaged in social roles, were married, were parents, or held jobs. Moreover, patients were more impaired in overall functioning, specifically in the domains of social/leisure, extended family, and marital roles than controls. Impairments in most roles were correlated with greater severity of symptoms, but not with degree of cognitive impairment, social environment, or background characteristics. The SAS-M is a useful addition to psychosocial batteries; however, the self-report format may not reflect others' perception of functioning. PMID- 9416650 TI - The association between adaptive and cognitive deficits in geriatric chronic schizophrenic patients. AB - Cognitive impairments have been shown to predict impairments in adaptive functioning in patients with chronic schizophrenia and to be more predictive of overall outcome than positive or negative symptoms of the illness. Both adaptive and cognitive impairments are multidimensional, and it is possible that correlations between these domains may be limited to certain aspects of these functions. In this study, 208 geriatric patients with chronic schizophrenia were examined with a cognitive battery and assessed with a structured scale to determine the extent of their adaptive functions. Instrumental and social skills deficits were more strongly correlated with cognitive impairments than with the severity of undercontrolled behavior. Each of the cognitive measures was correlated with global social-adaptive deficits, with minimal variation in the magnitude of correlations. These results suggest that interventions should be individually targeted to cognitive-adaptive impairments and undercontrolled behavior. PMID- 9416651 TI - Validity of specific subscales of the positive and negative symptom scales in older schizophrenia outpatients. AB - We investigated the construct validity of subscales of the Scale for the Assessment of Positive Symptoms (SAPS) and the Scale for the Assessment of Negative Symptoms (SANS) along with other measures of psychopathology in 109 schizophrenia outpatients aged 45-84 years. Scores on subscales of the SAPS, SANS and Brief Psychiatric Rating Scale (BPRS) and on the Hamilton Depression Scale (HAM-D) were subjected to a principal components analysis and orthogonal rotation followed by an extension analysis. In both analyses, three of four SAPS subscales had their highest loading on the positive symptom factor and four of five SANS subscales had their highest factor loading on the negative symptom factor. The SAPS bizarre behavior subscale, however, had a much higher loading on the depressive symptom factor than on the positive symptom factor, and the SANS avolition-apathy subscale had moderate loadings on both the negative symptom factor and the depressive symptom factor. The use of SAPS and SANS subscales to represent two constructs was largely (but not entirely) validated among middle aged and elderly schizophrenia outpatients. The SAPS bizarre behavior subscale and, to a lesser extent, the SANS avolition-apathy subscale appear to represent in this older population a separate construct which may be related to depressive symptoms. PMID- 9416652 TI - Therapeutic targets in late-life psychoses: review of concepts and critical issues. AB - Psychoses in late life are a diagnostic challenge because of disagreement over how these entities should be classified. The main diagnostic categories of late life psychoses include dementia with psychotic symptoms, late-onset schizophrenia, delusional disorder, early-onset psychotic disorders extending through late life, late-onset mood disorders, psychotic disorders caused by medical conditions or medications, and delirium. First onset of psychotic symptoms in late life is commonly associated with identifiable structural brain abnormalities and reflects underlying brain pathology. We reviewed the available literature on late-life psychotic manifestations, focusing on diagnostic classification and treatment approaches. Antipsychotics are the mainstay of treatment for these conditions, but should be used cautiously in elderly patients because of their increased sensitivity to side effects. Overall, appropriate research data on the effectiveness of various antipsychotic agents for late-life psychotic conditions are lacking. Non-antipsychotic psychotropic medications may be of value in managing some of these conditions. PMID- 9416653 TI - Neuroleptic dose reduction in older psychotic patients. AB - We conducted a non-randomized, rater-blind study to safely determine the lowest effective neuroleptic dosage in older psychotic patients and to evaluate the clinical, neuropsychological, and psychosocial effects of neuroleptic dosage reduction. Twenty-seven carefully selected patients with schizophrenia and related psychotic disorders over the age of 45 had their dosage tapered by 25% each month to determine their lowest effective dosage. These patients were compared with patients similar in age, gender, and education who were currently off neuroleptics (n = 19) or maintained on neuroleptics (n = 22). All groups were followed for 11 months. Over the follow-up period, 29% of patients in the taper group, 8% of neuroleptic-free patients, and 0% of patients in the maintenance group experienced some increase in psychopathology, although there was no significant change in mean PANSS score in any group, and no patient required hospitalization. Patients in the taper group were maintained on approximately 60% of their original neuroleptic dosage after restabilization. Extrapyramidal symptoms continued to improve over time in the taper group. Neuropsychological testing did not change significantly over time except for those in the taper group who experienced a decrease in memory-retention on the Hopkins Verbal Learning Test and a significant improvement in digit vigilance and Stroop Interference Index. Carefully selected middle-aged and elderly psychotic patients can have their neuroleptic medications reduced without a significant change in psychopathology. Extrapyramidal symptoms may continue to improve gradually over time. The impact on cognition functioning needs further investigation. PMID- 9416654 TI - New antipsychotic medications: strategies for evaluation and selected findings. AB - The unprecedented level of activity in the development of new antipsychotic medications can be traced to the 1989 approval of clozapine by the US Food and Drug Administration for treatment of refractory schizophrenia. This has encouraged the development of other new agents that share some of clozapine's receptor binding characteristics. A wide range of clinical trial designs are being used during the development of new antipsychotic medications. This article describes both basic designs and more innovative ones: flexible-dose designs that include placebo and conventional neuroleptic agents as controls; fixed-dose designs with multiple doses of experimental medication; and fixed-dose designs with multiple doses of the experimental and comparator medication. The strengths and weaknesses of each are identified. The need for long-term maintenance studies of newer agents is emphasized because psychotic disorders in general, and schizophrenia in particular, are chronic relapsing illnesses. The current status of four newer antipsychotic medications is considered: clozapine, risperidone, olanzapine, and sertindole. The importance of direct comparison among the newer antipsychotic medications in both short- and long-term trials is highlighted. PMID- 9416655 TI - The future of clinical research in mental disorders of late life. AB - After years of neglect, clinical research into late-life mental disorders is receiving the attention it deserves. Geriatric psychiatry is a growing field and the insights gained from research in this area have improved the diagnosis and treatment of mental illness in older persons. The challenge for the future is to continue to maintain a leadership role in mental health while working within the constraints of the managed care system in the United States. PMID- 9416657 TI - Imaging anatomy of the normal orbit. AB - Sectional imaging by CT and MRI now displays the gross anatomy of the orbit in vivo. Gross anatomy, axial and coronal CT, and MRI will be used to review the anatomic features of the orbit critical to image interpretation. Familiarity with the positions, relationships, and normal appearance of the major structures described and illustrated in this article will provide a solid basis for interpreting images of orbital pathology, as discussed in subsequent articles. PMID- 9416656 TI - An ophthalmic surgeon's view of orbital imaging techniques. AB - Selection of an orbital imaging technique requires a thorough understanding of pertinent anatomy applied to relevant clinical history and detailed ophthalmic examination. The clinical finding should direct the clinician to the imaging study that provides maximum information and narrows diagnostic considerations for the individual patient. Clinical examples are provided to illustrate the rationale in ordering magnetic resonance images, computed tomography, ocular ultrasound, and color Doppler arteriography of orbital processes. PMID- 9416658 TI - Orbital imaging techniques. AB - In the past 20 years, CT and MR imaging have emerged as the primary modalities for the evaluation of orbital disease. One can safely say that plain films have no role in the detailed evaluation of disorders of this morphological area. Given the bony enclosure, small size, soft tissue and fat content of this region, as well as globe movement, obtaining high-quality images of the orbit can be technically challenging. Our aim in this article, is to provide specific parameters for properly evaluating the orbit with CT and MRI. PMID- 9416659 TI - The pathological globe: clinical and imaging analysis. AB - Categorizing globe lesions based on clinical presentation can suggest a short list of diagnostic possibilities. Imaging of the globe with ultrasound, CT and MRI is presented with a focus on key differential points. The radiologist can then efficiently tailor the examination in order to differentiate among the diagnostic possibilities. PMID- 9416660 TI - Orbital trauma. AB - The globe and orbit constitute a very small portion of the body; however, trauma to this region assumes critical importance due to the high value we place on vision. The evaluation of orbital trauma has progressed rapidly with the development and wide distribution of computer-assisted imaging. Plain radiography, angiography, computed tomography (CT), and magnetic resonance imaging (MRI), may all be used in the evaluation of orbital trauma and the search for foreign bodies. PMID- 9416661 TI - Orbital infections. AB - Orbital infections account for the majority of primary intraorbital disease processes. Sinusitis is the most common etiology. Five stages of cellulitis secondary to sinusitis have been described. Systemic conditions which predispose to orbital infection include diabetes, septicemia, malignancy, and immunosuppresion. Clinical signs and symptoms include superficial inflammatory changes, as well as proptosis, limitation of extraocular motility, and visual loss. Causative agents are most commonly bacteria, with fungus, viruses, and parasites seen less frequently. Imaging is performed by CT and/or MRI which are complementary in certain cases. Differential diagnosis of imaging abnormalities includes inflammatory and granulomatous diseases, as well as neoplasm. PMID- 9416662 TI - Connections between neuroscientists from Europe and Latin America reinforced. PMID- 9416663 TI - The first confluence of mediterranean neuroscience. PMID- 9416664 TI - The brain has a body: adaptive behavior emerges from interactions of nervous system, body and environment. AB - Studies of mechanisms of adaptive behavior generally focus on neurons and circuits. But adaptive behavior also depends on interactions among the nervous system, body and environment: sensory preprocessing and motor post-processing filter inputs to and outputs from the nervous system; co-evolution and co development of nervous system and periphery create matching and complementarity between them; body structure creates constraints and opportunities for neural control; and continuous feedback between nervous system, body and environment are essential for normal behavior. This broader view of adaptive behavior has been a major underpinning of ecological psychology and has influenced behavior-based robotics. Computational neuroethology, which jointly models neural control and periphery of animals, is a promising methodology for understanding adaptive behavior. PMID- 9416665 TI - The functional neuroanatomy of episodic memory retrieval. PMID- 9416666 TI - The 'amyloid cascade hypothesis' of AD: decoy or real McCoy? PMID- 9416667 TI - Animal electricity as part of the 'one great whole'. PMID- 9416668 TI - The body in the brain: neural bases of corporeal awareness. AB - Recent studies have begun to unravel the brain mechanisms that underlie the mental representation of the body. Imitation of movements by neonates suggests an implicit knowledge of the body structure that antedates the adult body schema. This can include inanimate objects that bear systematic relations to the body, as shown by the elimination from self awareness of a body part and its associated paraphernalia after selective brain lesions. Dynamic aspects of the body schema are revealed by spontaneous sensations from a lost body part as well as by orderly phantom sensations elicited by stimulation of body areas away from the amputation line and even by visual stimulation. The mechanisms of the body schema exhibit stability, since some brain regions seem permanently committed to representing the corresponding body parts in conscious awareness, and plasticity, since brain regions deprived of their natural inputs from a body part become reactive to inputs from other body parts. PMID- 9416669 TI - Mechanisms of CNS response to systemic immune challenge: the febrile response. AB - The acute-phase reaction is the multisystem response to acute inflammation. The central nervous system (CNS) mediates a coordinated set of autonomic, endocrine and behavioral responses that constitute the cerebral component of the acute phase reaction. However, the mechanisms of immune signaling of the CNS remain controversial. Emerging evidence indicates that different parts of the acute phase reaction are initiated by distinct mechanisms and in different brain regions. Cytokines produced as a result of local infections (for example, in the abdominal or thoracic cavities) might activate vagal sensory fibers, resulting in sickness behavior and fevers. Additionally, circulating immune stimuli might activate meningeal macrophages and perivascular microglia along the borders of the brain, eliciting the local production of prostaglandins and responses such as fever, anorexia, sleepiness, and activation of the hypothalamo-pituitary-adrenal (HPA) axis. The biological importance of these responses might favor the existence of multiple parallel CNS pathways that are engaged by cytokines. PMID- 9416670 TI - Reactive astrocytes: cellular and molecular cues to biological function. AB - For several decades, the reactive gliosis that occurs after an injury to the CNS has been considered one of the major impediments to axonal regeneration. Nevertheless, recent studies have suggested that in certain conditions, reactive astrocytes may provide a permissive substratum to support axonal regrowth. The important criteria, allowing for the distinction between permissive and non permissive gliosis, are the ultrastructural 3D organization of the scar and more importantly the recognition molecules expressed by reactive astrocytes. Reactive astrocytes express surface molecules and produce various neurotrophic factors and cytokines. The latter in turn might modulate the production of recognition molecules by reactive astrocytes, allowing them to support post-lesional axonal regrowth. Although numerous recent articles have focused on cytokines and cell adhesion molecules, scant attention has been paid to reactive astrocytes. Reactive astrocytes should be considered a key element, like neurons, of a dynamic environment, thus forming with neurons a functional unit involved in homeostasis, plasticity and neurotransmission. Attempts are in progress to identify molecular markers for reactive astrocytes. PMID- 9416672 TI - Principles of acoustic motion detection in animals and man. AB - Motion provides one of the most important cues for survival, because it helps to break the camouflage of a predator or a prey and because it allows predictions about the future path of an object. Recent data on the processing of acoustic motion have yielded some astonishing findings, suggesting that the psychophysical, neurological and neurophysiological mechanisms underlying the detection and representation of acoustic motion are quite similar to those underlying the detection and representation in other modalities, especially in vision. A further comparison of these similarities and differences with respect to the different environmental constraints posed for the different modalities may help in understanding general problems associated with motion computations. PMID- 9416671 TI - Molecular biology of insect neuronal GABA receptors. AB - Ionotropic gamma-aminobutyric acid (GABA) receptors are distributed throughout the nervous systems of many insect species. As with their vertebrate counterparts, GABAA receptors and GABAC receptors, the binding of GABA to ionotropic insect receptors elicits a rapid, transient opening of anion-selective ion channels which is generally inhibitory. Although insect and vertebrate GABA receptors share a number of structural and functional similarities, their pharmacology differs in several aspects. Recent studies of cloned Drosophila melanogaster GABA receptors have clarified the contribution of particular subunits to these differences. Insect ionotropic GABA receptors are also the target of numerous insecticides and an insecticide-resistant form of a Drosophila GABA-receptor subunit has enhanced our understanding of the structure-function relationship of one aspect of pharmacology common to both insect and vertebrate GABA receptors, namely antagonism by the plant-derived toxin picrotoxinin. PMID- 9416673 TI - The complex structure of a simple memory. AB - Operant conditioning of the vertebrate H-reflex, which appears to be closely related to learning that occurs in real life, is accompanied by plasticity at multiple sites. Change occurs in the firing threshold and conduction velocity of the motoneuron, in several different synaptic terminal populations on the motoneuron, and probably in interneurons as well. Change also occurs contralaterally. The corticospinal tract probably has an essential role in producing this plasticity. While certain of these changes, such as that in the firing threshold, are likely to contribute to the rewarded behavior (primary plasticity), others might preserve previously learned behaviors (compensatory plasticity), or are simply activity-driven products of change elsewhere (reactive plasticity). As these data and those from other simple vertebrate and invertebrate models indicate, a complex pattern of plasticity appears to be the necessary and inevitable outcome of even the simplest learning. PMID- 9416674 TI - Displacement of the tubular genital tract of the ewe during pregnancy. AB - The displacement of the tubular genital tract of ewes during pregnancy, in particular of the cervix relative to the cranial pelvic brim, was studied by sequential radiography using radio-opaque markers attached surgically before tupping. In the first year, 13 two-year-old parous ewes were fed a complete pelleted diet containing either 15 or 25 per cent fibre throughout pregnancy. The distances between the markers attached to the tubulogenital tract increased and the distance between the cervix and the cranial pelvic brim increased slightly with advancing pregnancy; the diet had no effect. In the second year, only cervical displacement was studied in 11 ewes that had suffered a cervicovaginal prolapse the previous year, and in five of their female progeny. They were divided into two groups, one of which was fed a 15 per cent-fibre complete pelleted diet and the other hay ad libitum throughout pregnancy. In two ewes, one from each dietary group, the prolapse recurred less than two weeks from term; initially it was a stage 2 prolapse, where the vaginal wall fails to return to its normal position when the ewe stands, but in one ewe it became a stage 3 prolapse, with the vagina completely everted and the cervix visible. During the stage 2 prolapse there was no evidence of caudal displacement of the cervix, whereas during the progression to the stage 3 prolapse the cervix became displaced 33 cm caudally over two to three hours. PMID- 9416675 TI - Antibodies to Brucella in marine mammals around the coast of England and Wales. AB - Following the isolation of previously unrecognised species of Brucella from stranded seals and cetaceans in Scotland and northern England, a serological survey was carried out to investigate the range of marine mammal species which may have been exposed to Brucella species around the coasts of England and Wales, the prevalence of infection and the temporal and geographical distribution of seropositive animals. Serum collected from 153 stranded marine mammals from the coasts of England and Wales between 1989 and 1995 were tested by competitive and indirect ELISA. Positive titres were recorded for six of 62 (10 per cent) grey seals (Halichoerus grypus), one of 12 (8 per cent) common seals (Phoca vitulina), 11 of 35 (31 per cent) harbour porpoises (Phocoena phocoena) and nine of 29 (31 per cent) common dolphins (Delphinus delphis) tested. Positive titres were also found in a striped dolphin (Stenella coeruleoalba), a bottlenose dolphin (Tursiops truncatus), a killer whale (Orcinus orca) and a pilot whale (Globicephala melas). The seropositive animals were from all around the coasts of England and Wales and the first seropositive sample was from a common dolphin in 1990. PMID- 9416676 TI - Isolation of mycobacteria from lymph node lesions in deer. AB - A total of 14,842 farmed deer were slaughtered and examined postmortem in Irish abattoirs between January 1993 and September 1996. Lymph node lesions were detected in 119 deer and these were examined histopathologically and cultured. A total of 115 of the lesions were characterised as tuberculous and, on culture, Mycobacterium avium was isolated from 49 of them, M bovis from 41, unclassified mycobacteria from five, and 20 of the tuberculous lesions did not yield any isolate. Tubercles which yielded M avium on culture contained on average more acid-fast bacilli, more epithelioid macrophages and fewer Langhans giant cells than tubercles from which M bovis was isolated. Twelve lesions from feral deer culled from a national park were also examined and M bovis was isolated from nine. PMID- 9416677 TI - Oxidant stress in the equine lung: response to oral prednisolone. PMID- 9416678 TI - Acute haemorrhagic pancreatitis associated with canine visceral leishmaniasis. PMID- 9416679 TI - Vestibulo-vaginal hypoplasia in a mare. PMID- 9416680 TI - Influence of cumulus cells on in vitro maturation of denuded buffalo oocytes. PMID- 9416681 TI - Ultrasonographic examination of the abomasum of neonatal lambs. PMID- 9416682 TI - Screening for caseous lymphadenitis. PMID- 9416683 TI - Rabies and quarantine: serological testing. PMID- 9416684 TI - Recall of droplix. PMID- 9416685 TI - Left displaced abomasum in sheep. PMID- 9416686 TI - Oral fibropapillomas in young cats. PMID- 9416687 TI - Is Epstein-Barr virus a human oncogene or only an innocent bystander? PMID- 9416688 TI - The spectrum of multiple myeloma: diagnostic and biological implications. AB - Myeloma is a common and well-studied hematopoietic neoplasm with an impressive spectrum of clinical, laboratory, and histological findings. To enhance our understanding of the diversity of myeloma, including its earliest forms, the clinical and pathological findings in 145 cases of myeloma were documented and analyzed. Our analysis indicated that myeloma has at least two distinct subtypes: one presenting with bone lesions and a nodular growth pattern and the other presenting with anemia and an infiltrative growth pattern. The relationship of these two forms to plasma cell biology is not clear, although both types appear to arise in the marrow. The criteria used in this study identified 85% of cases overall, with a range of 70% to 100%, depending on clinical presentation. Immunoperoxidase studies are required to establish the diagnosis in patients with early marrow involvement. Myeloma in younger patients appears to be clinically and pathologically similar to myeloma in older patients. Factors such as dysplasia, immunoglobulin type, or leukemic phase were evenly distributed among clinical presentations and did not apparently identify clinicopathological subtypes of myeloma. PMID- 9416690 TI - The gastric hypercellular microleiomyoma as a precursor lesion for clinical gastrointestinal stromal tumors. AB - Differentiation features and proliferation activity of 67 gastric microleiomyomas (microLMs) and 53 clinical gastrointestinal stromal tumors (GISTs) of the stomach were compared. The 67 microLMs were divided into two categories on the basis of cellularity: 53 hypocellular and 14 hypercellular types, and the 39 GISTs were divided into 13 low-grade and 40 high-grade lesions. Immunohistochemically, 49 hypocellular microLMs (92%) were positive for alpha-smooth muscle actin and desmin, whereas only 16 (30%) were stained for vimentin. Conversely, all 14 hypercellular microLMs were positive for vimentin, and only one (7%) was positive for alpha-smooth muscle actin and desmin. Five low-grade (38%) and 14 high-grade GISTs (35%) were positive for alpha-smooth muscle actin, and 12 low-grade (92%) and all 40 high-grade GISTs were stained for vimentin. CD34 was positive in 10 hypocellular microLMs (19%), all 14 hypercellular microLMs, 10 low-grade GISTs (77%), and 38 high-grade GISTs (95%). Hypercellular microLM thus showed similarities to clinical GIST and also exhibited significantly higher proliferation activity than hypocellular microLM on analysis of the Ki-67 labeling index and argyrophilic nucleolar organizer regions staining. The findings indicate that the hypercellular microLM may be a direct precursor for clinical GIST, both showing a primitive mesenchymal cell nature. PMID- 9416689 TI - Intermediate filament expression by normal and diseased human corneal epithelium. AB - Cicatricial conjunctivitis may be a sequel to systemic disorders (eg, Stevens Johnson syndrome, cicatricial pemphigoid) or local disorders such as chemical burns. The cicatrisation is often associated with corneal epithelial changes that cause visual loss. These have been attributed to encroachment of the conjunctival epithelium over the cornea. However, the epithelial anomalies are poorly understood. We investigated the corneal epithelial changes in cicatricial conjunctivitis with an immunohistochemical study of intermediate filaments in normal and pathological specimens. Our results show that the normal corneal epithelium is immunoreactive for cytokeratin 3 (CK 3) but not cytokeratin 19 (CK 19), whereas normal conjunctival epithelium is CK 3 negative and CK 19 positive. Conjunctiva artificially transposed over the cornea (after therapeutic conjunctival flap reconstruction) retained the normal pattern of conjunctival cytokeratin expression (CK 3 negative, CK 19 positive). Conversely, the entire corneal epithelium exhibited the normal cytokeratin pattern (CK 3 positive, CK 19 negative) in 82% of Stevens-Johnson, 80% of cicatricial pemphigoid, and 69% of chemical burns specimens. The findings suggest that conjunctival encroachment is not responsible for the changes at the corneal surface in cicatricial conjunctivitis and that the abnormal corneal epithelium is derived from native corneal cells in these diseases. PMID- 9416691 TI - Analysis of p53 expression in osteosarcoma of the jaw: correlation with clinicopathologic and DNA ploidy findings. AB - The objective of this study was to determine whether p53 expression correlated with clinicopathological and DNA ploidy in osteosarcoma of the jaw, a particular subtype of osteosarcoma associated with an older age at presentation, longer median survival, rare metastasis, and death due to uncontrolled local recurrence. Seventeen cases of osteosarcoma of the jaw were stained for p53 using CM1 antibody (Novocastra). Eight cases (47.5%) showed nuclear staining in more than 10% of the neoplastic nuclei. Ploidy analysis showed 13 aneuploid (eight p53+) and three diploid (none p53+) tumors. No significant relationship was found between p53 positivity and clinicomorphological features, DNA ploidy or survival. p53-positive cases were, however, more commonly associated with aneuploidy and chondroblastic differentiation. These results indicate that p53 immunostaining does not seem to provide prognostic information in osteosarcoma of the jaw. PMID- 9416692 TI - Abnormal intracellular and extracellular distribution of basement membrane material in papillary carcinoma and hyalinizing trabecular tumors of the thyroid: implication for deregulation of secretory pathways. AB - Papillary carcinoma (PC) and hyalinizing trabecular tumors (HTT) of the thyroid share several morphological features, including the presence of nuclear pseudoinclusions (NPI). One of the distinct characteristics of HTT is its hyalinizing stroma, which contains abundant basement membrane (BM) material. We investigated the distribution of BM material in PC and HTT. Fifteen cases of PC and nine cases of HTT were analyzed immunohistochemically with monoclonal antibodies for type IV collagen and laminin. Three stromal staining patterns were observed: (1) linear staining along the epithelium lining papillae, between trabeculae, and around follicles; (2) focal absence of staining; (3) lumpy or diffuse stromal staining. Although the latter was more commonly seen in HTT, all three patterns were present in both tumor types. More interestingly, we observed two hitherto undescribed intracellular staining patterns in both tumor types: intracytoplasmic dotlike staining and staining of NPI. Electron microscopy was performed in three cases of PC. Dilated cisternae of endoplasmic reticulum containing dense amorphous material resembling BM were observed in the cytoplasm in one case and in the NPI in another. These findings suggest the presence of a common pathway for the abnormal production of BM in both PC and HTT. Two mechanisms that may account for the abnormal intracellular detection of BM materials are proposed: (1) intracellular invagination/phagocytosis of extracellular matrix by the tumor cells; (2) abnormal production or alteration in secretory pathway in tumor cells resulting in intracellular accumulation and intranuclear invagination. The combination of immunohistochemical and electron microscopic findings favors the latter. The similar patterns of BM deposition shared by PC and HTT further support the hypothesis that PC and HTT are related to each other. PMID- 9416694 TI - Cytogenetic findings in phyllodes tumors of the breast: karyotypic complexity differentiates between malignant and benign tumors. AB - Clonal karyotypic abnormalities were detected in short-term cell cultures from six phyllodes tumors of the breast. Whereas all five benign tumors had simple chromosomal changes, the highly malignant one had a near-triploid stemline, indicating that karyotypic complexity is a marker of malignancy in phyllodes tumors. Interstitial deletions of the short arm of chromosome 3, del(3)(p12p14) and del(3)(p21p23),were the only aberrations in two benign tumors. Cytogenetic polyclonality was detected in three benign tumors: two had cytogenetically unrelated clones, whereas the third had three different, karyotypically related cell populations as evidence of clonal evolution. The finding of clonal chromosome abnormalities in both the epithelial and connective tissue components of the phyllodes tumors indicates that they are genuinely biphasic, that is, that both components are part of the neoplastic parenchyma. PMID- 9416693 TI - Utility of 123C3 monoclonal antibody against CD56 (NCAM) for the diagnosis of small cell carcinomas on paraffin sections. AB - CD56 is immunohistochemically detectable in virtually all small cell carcinomas on frozen sections. The authors retrospectively tested the usefulness of the monoclonal antibody 123C3 against CD56 to differentiate pulmonary and extrapulmonary small cell carcinomas from nonneuroendocrine non-small cell carcinomas by paraffin-section immunohistochemistry after antigen retrieval. The study included 70 small cell carcinomas and 344 primary and metastatic nonneuroendocrine carcinomas of various primary sites. The staining results were compared with specific neuroendocrine markers (CD57, Chromogranin A, Synaptophysin). The monoclonal antibody 123C3 diffusely stained most small cell carcinomas with a strong membranous pattern (sensitivity: 0.99). The staining intensity was not diminished in areas with crush artifacts or after decalcification. The neuroendocrine markers had a combined sensitivity of only 0.44 for small cell carcinomas. With regard to nonneuroendocrine carcinomas, the 123C3 antibody stained 7 of 28 ovarian carcinomas, 6 of 30 renal cell carcinomas, 2 of 10 endometrial carcinomas, two of three nonneuroendocrine large cell carcinomas of the lung, 1 of 38 adenocarcinomas, and 4 of 52 squamous cell carcinomas of the lung. Urothelial carcinomas, hepatocellular carcinomas, squamous carcinomas of the head/neck and cervix uteri, as well as adenocarcinomas of the breast, stomach, colon, pancreas, and prostate, showed no immunoreactivity for CD56. The specificities of 123C3 and the combined neuroendocrine markers for small cell carcinomas were 0.94 and 0.95, respectively. The authors conclude that monoclonal antibody 123C3 might be useful for the immunohistochemical differentiation of small cell carcinomas from nonneuroendocrine carcinomas on paraffin sections, especially in small and crushed biopsy specimens. PMID- 9416695 TI - Alveolar lipoproteinosis in lung allograft recipients. AB - Three cases of pulmonary alveolar proteinosis developing in lung allograft recipients are reported. In each case, repeated bouts of alveolar damage from harvest/reperfusion injury, rejection, and infection were observed before the development of intraalveolar accumulation of granular, periodic acid-Schiff positive material in the allograft lungs. It is speculated that iatrogenic immunosuppression combined with defective clearance of alveolar material by alveolar macrophages created a milieu conducive to the accumulation of surfactant, lipoprotein, and fibrinous debris that was morphologically identical to alveolar proteinosis. PMID- 9416696 TI - Value of inhibin in the identification of granulosa cell tumors of the ovary. AB - Inhibins are peptide hormones that participate in the regulation of the pituitary gonadal feedback system and are selectively expressed by cells of sex cord stromal derivation. To determine the efficacy of this marker for distinguishing granulosa cell tumors, 134 primary and metastatic lesions of the ovary were evaluated for expression of the alpha-subunit of inhibin in routinely processed formalin-fixed, paraffin-embedded tissue. A variety of sex cord-stromal tumors (SCST), including 35 adult and juvenile granulosa cell tumors, 14 fibroma thecomas, and 18 other sex cord-stromal proliferations, were studied. In addition, 33 surface epithelial neoplasms, 12 germ cell tumors, 11 metastases, and 11 miscellaneous ovarian neoplastic proliferations were evaluated. Among the non-granulosa cell neoplasms, special emphasis was placed on primary neoplasms and metastases that histologically simulated granulosa cell tumors. Thirty-three of 35 (94%) granulosa cell tumors were immunoreactive compared with 2 of 12 (17%) primary ovarian endometrioid tumors, one of nine (11%) primary ovarian transitional cell (Brenner) proliferations, and 3 of 17 (18%) other primary and metastatic poorly differentiated (undifferentiated) carcinomas. In 31 of the 35 granulosa cell tumors, inhibin staining was of moderate to strong intensity or was present in at least half of the constituent cells, whereas only 2 of 33 primary surface epithelial neoplasms fulfilled the same criteria, showing weak staining of 70% to 80% of the cells. In contrast, 10 of 14 (71%) ovarian fibroma thecomas and 17 of 18 (94%) other sex cord-stromal proliferations were positive for inhibin. Nonneoplastic luteinized stromal cells stained for inhibin in 29 of 85 cases in which they could be evaluated. The results of this study show that although it is not completely specific and cannot reliably distinguish granulosa cell tumors from fibroma-thecomas or other ovarian sex cord-stromal proliferations, inhibin can be used to help distinguish sex cord-stromal neoplasms from most primary and metastatic non-SCST. Caution should be exercised in the interpretation of inhibin-positive cells, because a wide variety of primary and metastatic ovarian tumors may contain significant numbers of positively staining luteinized cells. PMID- 9416697 TI - Phenotypic and genotypic characteristics of aberrant crypt foci in human colorectal mucosa. AB - Aberrant crypt foci (ACF) in colorectal mucosa are proposed to be the earliest morphological lesion in the development of neoplasia, but their characteristics remain controversial. We therefore studied the epithelial phenotype and genotype of ACF from patients with familial adenomatous polyposis (FAP) and of sporadic ACF by evaluating glycoprotein markers associated with neoplasia (lectins Dolichus biflorus agglutinin and peanut agglutinin; monoclonal antibody CA 19-9 against sialyl Lewis-a blood group substance), expression of proliferating cell nuclear antigen, and ras proto-oncogene mutations. The utility of the markers was established by comparing adenomas and hyperplastic polyps. Most FAP ACF resembled adenomas and were found to differ from sporadic ACF in their high frequency of dysplasia, staining with Dolichus biflorus agglutinin, expression of sialyl Lewis a, proliferation in the epithelium of upper crypts, and low frequency of ras gene mutations (P = .04 to < .0000001). By contrast, sporadic ACF and a subset of FAP ACF had phenotypic characteristics resembling hyperplastic polyps but usually had ras mutations, which were inversely related to dysplasia (P = .00009). Our findings suggest that "aberrant crypt focus" is a generic term analogous to "polyp" and requires further histopathologic, phenotypic, or genotypic classification into dysplastic and heteroplastic (hetero = other, plasia = form) types. Dysplastic ACF represent potential precursors to colorectal adenomas and adenocarcinomas, but heteroplastic ACF appear to be associated, rather than precursor, lesions. PMID- 9416698 TI - Deletion of Epstein-Barr virus latent membrane protein 1 gene in United States and Brazilian Hodgkin's disease and reactive lymphoid tissue: high frequency of a 30-bp deletion. AB - A 30-basepair (bp) deletion in the Epstein-Barr virus (EBV) latent membrane protein 1 (LMP1) gene has been reported in nasopharyngeal carcinoma and EBV associated malignant lymphomas. Prior studies have found the deletion in about 10% to 28% of cases of Hodgkin's disease (HD), particularly in cases with aggressive histology. We studied the prevalence of 30-bp LMP1 gene deletion in EBV-positive HD in the United States (US) (12 cases) and Brazil (26 cases) with comparison to reactive lymphoid tissues (21 cases) and HD without EBV-positive Reed-Sternberg cells (15 cases). We studied the status of the LMP1 gene by Southern blot hybridization of polymerase chain reaction (PCR) products obtained after amplification with primers spanning the site of the deletion. We also performed EBV typing, EBER1 in situ hybridization, and LMP1 protein immunohistochemistry. EBV was detected in 12/26 (46%) cases of HD from the US and 26/27 (96%) cases of Brazilian HD. The 30-bp LMP1 gene deletion was observed in 4/12 (33%) cases of EBV-positive HD from US, and 12/26 (46%) cases of Brazilian EBV-positive HD, including 3 cases of type B EBV, as compared with 12/21 (57%) reactive lymphoid tissues and 9/15 (60%) cases of EBV-negative HD. US and Brazilian HD showed a higher prevalence of the 30-bp LMP1 gene deletion, compared with studies of others. The unexpected finding of high incidence of 30-bp deletion in LMP1 gene in reactive lymphoid tissue and HD without EBV-positive Reed-Sternberg cells suggests that this deletion may not be relevant to HD pathogenesis in most cases. PMID- 9416699 TI - Detection of Epstein-Barr virus transcripts in anaplastic large-cell lymphomas by mRNA in situ hybridization. AB - Anaplastic large-cell lymphoma (ALCL) is a recently proposed subset of non Hodgkin's lymphoma. To determine whether Epstein-Barr virus (EBV) is associated with this lymphoma, we performed mRNA in situ hybridization on seven cases of ALCL using a probe consisting of an RNA sequence complementary to the transcripts of BamHIW fragment of the EBV genome. We detected BamHIW transcripts of EBV in the majority of atypical large cells of all cases of ALCL, but in none of three cases of lymphoblastic and small lymphocytic lymphomas. Furthermore, we detected latent membrane protein-1 (LMP1) in two cases of ALCL by means of immunofluorescence and immunoperoxidase stainings. These findings suggest that EBV is involved in the neoplastic transformation for ALCL as in the case of Hodgkin's disease, which shares several clinicopathologic features with ALCL. PMID- 9416700 TI - Lymphangioleiomyomatosis: recurrence after single lung transplantation. AB - Lymphangioleiomyomatosis (LAM) is a rare disease which afflicts young women of childbearing age. Recently, it has been listed as an indication for lung transplantation. We describe a case of recurrent LAM in a 31-year-old woman occurring in the allograft of a male donor after single lung transplantation. Nonisotopic in situ hybridization shows that the smooth muscle cell proliferation is of donor origin. PMID- 9416701 TI - Invasion and destruction of mucosal plasma cells by Tropheryma whippelii. AB - Whipple's disease is a poorly understood systemic disorder associated with the bacillus, Tropheryma whippelii. An early stage of Whipple's disease is studied by using electron microscopy (ELMI) and immunohistochemistry. The diagnosis was confirmed by polymerase chain reaction-amplification and sequencing of the 16S ribosomal-RNA of the bacterium. By using ELMI, Tropheryma whippelii was found in plasma cells and macrophages in the jejunal mucosa. The immunoglobulin (Ig)A positive plasma cells were focally destructed and their number significantly reduced. However, the bacilli in the plasma cells were morphologically intact. In contrast, the macrophages showed no signs of cell destruction, but contained bacilli in various stages of disintegration. A cytopathic effect of Tropheryma whippelii to IgA plasma cells may be the reason for the commonly found plasma cell reduction in the small intestine mucosa and an important pathogenic mechanism contributing to the evasion of the bacilli from local immune response. PMID- 9416702 TI - Adenomas of the rete ovarii. AB - This article reports the clinicopathological and immunohistochemical findings of two cases of adenoma of the the rete ovarii (RO), one unilateral and the other bilateral, presenting with atypical histological features in the right ovary. Both tumors were incidental findings in 62- and 64-year-old patients presenting with metrorrhagia. The predominantly cystic lesions measured 2 cm and 3 cm in diameter and microscopically, they were tubulopapillary proliferations of regular columnar cells with clear cytoplasm. The stroma showed extensive differentiation of polygonal, Leydig-like cells which was associated in both cases with simple endometrial hyperplasia. In both cases rete and hilar mesonephric remnants were found in the vicinity of the lesion. The atypical lesion in one case had a complex papillary proliferation different in pattern and cellularity from a retiform Sertoli-Leydig cell tumor. It showed extensive areas of eosinophilic change, pleomorphism, and a few mitoses but did not invade the adjacent ovarian stroma. Its stroma also had steroidally active cells. The patient was alive and well after a follow-up interval of 3 years. Immunohistochemically, the lesions were diffusely positive for CAM 5.2, vimentin, epithelial membrane antigen, OC 125, OC 125, and progesterone receptors. PMID- 9416703 TI - Epithelioid sarcoma producing granulocyte colony-stimulating factor. AB - An epithelioid sarcoma of the perineum of a 60-year-old man with widespread metastases produced leukocytosis, myeloid hyperplasia of the bone marrow, and splenomegaly. High titers of granulocyte colony-stimulating factor (G-CSF) were found in the patient's serum and primary culture medium of the tumor tissue. The tumor tissue extract contained m-RNA for G-CSF in large quantities, proving that the tumor was the source of this cytokine. PMID- 9416704 TI - Pulmonary intralobar sequestration in a patient with cystic fibrosis. AB - We report a case in which pulmonary Intralobar Sequestration (ILS) was an incidental finding at autopsy in an adult with Cystic Fibrosis. Two aberrant arteries from the descending thoracic aorta supplied a bronchial cystic lesion in the right lower lobe. Termination of the segmental bronchus and scar formation proximal to the cyst suggested prior bronchial obliteration. The elastic configuration of the aberrant aortic-derived vessels of the sequestration contrasted sharply with massively hypertrophied, muscular, bronchial arteries which supplied the bronchiectatic upper lobe. Sections of inferior pulmonary ligament were studied in five additional patients with CF but without ILS. Small muscular arteries were consistently noted within the inferior pulmonary ligament. These histologic findings support the concept that the vascular portion of ILS is congenital, whereas the bronchocystic component, in some cases, may be acquired. PMID- 9416705 TI - A link between hepatitis viruses and lymphoproliferative disorders. PMID- 9416706 TI - The data security aspects of telepathology via the Internet have been ignored. PMID- 9416707 TI - The "big little problem" of postoperative nausea and vomiting: do we know the answer yet? PMID- 9416708 TI - Intubation difficulty scale: anticipated best use. PMID- 9416709 TI - Drug distribution: less passive, more active? PMID- 9416710 TI - Efficacy, dose-response, and safety of ondansetron in prevention of postoperative nausea and vomiting: a quantitative systematic review of randomized placebo controlled trials. AB - OBJECTIVE: The authors reviewed efficacy and safety data for ondansetron for preventing postoperative nausea and vomiting (PONV). METHODS: Systematically searched, randomized, controlled trials (obtained through MEDLINE, EMBASE, Biological Abstracts, manufacturer's database, manual searching of journals, and article reference lists) were analyzed. Relevant end points were prevention of early PONV (within 6 h after surgery) and late PONV (within 48 h) and adverse effects. Relative benefit and number-needed-to-treat were calculated. The number needed-to-treat indicated how many patients had to be exposed to ondansetron to prevent PONV in one of them who would have vomited or been nauseated had he or she received placebo. RESULTS: Fifty-three trials were found that had data from 7,177 patients receiving 24 different ondansetron regimens and from 5,712 controls receiving placebo or no treatment. Average early and late PONV incidences without ondansetron were 40% and 60%, respectively. There was a dose response for oral and intravenous ondansetron. Best number-needed-to-treat to prevent PONV with the best documented regimens was between 5 and 6. This was achieved with an intravenous dose of 8 mg and an oral dose of 16 mg. Antivomiting efficacy was consistently better than antinausea efficacy. Efficacy in children was poorly documented. Ondansetron significantly increased the risk for elevated liver enzymes (number-needed-to-harm was 31) and headache (number-needed-to-harm was 36). CONCLUSIONS: If the risk of PONV is very high, for every 100 patients receiving an adequate dose of ondansetron 20 patients will not vomit who would have vomited had they received placebo. The antinausea effect is less pronounced. Of these 100, three will have elevated liver enzymes and three will have a headache who would not have had these adverse effects without the drug. PMID- 9416711 TI - The intubation difficulty scale (IDS): proposal and evaluation of a new score characterizing the complexity of endotracheal intubation. AB - BACKGROUND: A quantitative scale of intubation difficulty would be useful for objectively comparing the complexity of endotracheal intubations. The authors have developed a quantitative score that can be used to evaluate intubating conditions and techniques with the aim of determining the relative values of predictive factors of intubation difficulty and of the techniques used to decrease such difficulties. METHODS: An Intubation Difficulty Scale (IDS) was developed, based on parameters known to be associated with difficult intubation. It was then evaluated prospectively in a group of 311 consecutive prehospital intubations and 315 intubations in an operating room. In the operating room, the IDS was compared with two other parameters: the time to completion of intubation and the visual analog scale (VAS). Time was measured by an independent observer. Operators in both groups completed a checklist regarding the conditions of intubation. RESULTS: There is a good correlation between the IDS scale and the VAS assessment of difficulty and time to completion of intubation. VAS and time to completion have a significant but lesser correlation to each other. Comparison of IDS with operator-assessed subjective categorical impression of difficulty by Kruskall-Wallis was statistically significant. CONCLUSIONS: The IDS correlates with but is less subjective than the VAS and categorical classification. IDS correlates with time to intubation, but it offers details regarding the difficulty encountered that time alone does not. This score may not only aid in evaluation of factors linked to difficult intubations, but it may provide a uniform approach to comparing studies related to this subject. PMID- 9416712 TI - Greater incidence of delirium during recovery from sevoflurane anesthesia in preschool boys. AB - BACKGROUND: In the authors' clinical experience, preschool children are more likely to show delirium after sevoflurane than are older children. METHODS: Sixty three preschool boys aged 3-5 yr (classified as American Society of Anesthesiologists [ASA] physical status I), and 53 school-age boys aged 6-10 yr (ASA physical status I) who underwent minor urologic surgery were randomly assigned to receive either halothane or sevoflurane, thus creating four groups: preschool-halothane (n = 32), preschool-sevoflurane (n = 31), school-halothane (n = 27), and school-sevoflurane (n = 26). Anesthesia was induced by inhalation of halothane or sevoflurane in oxygen and was maintained at 1 minimum alveolar concentration of each agent throughout surgery. For intra- and postoperative analgesia, caudal block with 0.5-1.0 ml/kg 0.25% plain bupivacaine and topical infiltration with 3-5 ml 1% lidocaine were provided for all patients. Recovery characteristics and incidence of delirium on emergence were compared among the four groups. RESULTS: Two patients in the preschool-halothane group, one in the preschool-sevoflurane group, and one in the school-halothane group were excluded from the comparison because of insufficient analgesia or agitation before induction. In both age groups, the time to emergence from sevoflurane was significantly faster (about 3 min) than from halothane. The incidence of delirium during recovery in the preschool-sevoflurane group (40%) was significantly greater than that in the other groups (preschool-halothane, 10%; school halothane, 15.4%; school-sevoflurane, 11.5%). CONCLUSION: Sevoflurane provided quicker emergence and early recovery compared with halothane, but the incidence of delirium was greater in preschool boys after sevoflurane. PMID- 9416713 TI - Single-needle celiac plexus block: is needle tip position critical in patients with no regional anatomic distortions? AB - BACKGROUND: The "single-needle" celiac plexus block is becoming a popular technique. Despite different approaches and methods used to place the needle, the success of the block depends on adequate spread of the injectate in the celiac area. In the present retrospective study, the influence of needle tip position in relation to the celiac artery on injectate spread was evaluated. METHODS: Among 138 cancer patients subjected, via an anterior approach, to computed tomography (CT)-guided single-needle neurolytic celiac plexus block, a radiologist, blinded to the aim of the study, retrospectively selected 53 cases with normal anatomy of the celiac area as judged by CT. The decision was based on images obtained before the block. Patients were then classified into either group A (29 patients), in whom the needle tip was caudad to the celiac artery, and group B (24 patients), in whom it was cephalad. To evaluate CT patterns of neurolytic (mixed with contrast) spread, the celiac area was divided on the frontal plane into four quadrants: upper right and left and lower right and left, as related to the celiac artery. Patient assessments by visual analog scale were reviewed to evaluate the degree of pain relief. Pain relief 30 days after block was judged as long-lasting. The patterns of contrast spread in relation to the needle position and pain relief according to the number of quadrants with contrast were analyzed. RESULTS: The percentage of cases with four quadrants with contrast was higher when the needle tip was cephalad (58%, group B) than when it was caudad (14%, group A) to the celiac artery (P < 0.01). The percentage of patients with four and three quadrants with contrast was also higher in group B at 79% than in group A at 38% (P < 0.01). A significant difference in long-lasting pain relief was observed between patients with four quadrants with contrast (18 of 18, 100%; 95% confidence interval [CI], 81-100%) and patients with three quadrants with contrast (5 of 12, 42%; 95% CI, 15-72%) (P < 0.01). No patients showing two or one quadrant with contrast had long-lasting pain relief. CONCLUSIONS: These findings suggest that, when the celiac area is free from anatomic distortions, and the single-needle neurolytic celiac plexus block technique is used, the needle tip should be positioned cephalad to the celiac artery to achieve a wider neurolytic spread. It also appears that only a complete (four quadrants) neurolytic spread in the celiac area can guarantee long-lasting analgesia. PMID- 9416714 TI - Systemic absorption and block after epidural injection of ropivacaine in healthy volunteers. AB - BACKGROUND: For local anesthetics, the process of removal from the site of administration influences the duration of anesthesia and the risk for systemic toxicity to develop. The systemic absorption of epidural ropivacaine and the time profile of sensory and motor block were studied in healthy volunteers. METHODS: Nine persons simultaneously received 150 mg ropivacaine hydrochloride (7.5 mg/ml) epidurally and 40 mg deuterium-labeled (2H3)ropivacaine hydrochloride (0.25 mg/ml) intravenously. Peripheral arterial and venous plasma samples were collected, and assessments of sensory and motor block were made. RESULTS: The arterial plasma concentrations increased faster than the venous concentrations, with 50% higher maximum concentrations after both intravenous and epidural administration. The absorption was biphasic. A correlation was seen between the duration of sensory block and the slower absorption half-life; that is, the longer the half-life, the longer the duration. The extent of spread varied among the volunteers, with the median upper block level not exceeding T12. The motor block (Bromage score 1) was of slower onset (median, 0.4 h) and of shorter duration (median, 4.1 h) than the sensory block (onset, 0.2 h; duration, 6.5 h at L2 medians). CONCLUSIONS: As much as 50% differences were seen in the arteriovenous plasma concentrations of ropivacaine during the first hour, which has implications for the interpretation of systemic toxic plasma concentrations. The absorption into the general circulation was biphasic, with a correlation between the sensory block and the slower absorption half-life. A faster onset and a longer duration of sensory compared with motor block was seen. PMID- 9416715 TI - Mild intraoperative hypothermia prolongs postanesthetic recovery. AB - BACKGROUND: Intraoperative hypothermia is common and persists for several hours after surgery. Hypothermia may prolong immediate recovery by augmenting anesthetic potency, delaying drug metabolism, producing hemodynamic instability, or depressing cognitive function. Accordingly, the authors tested the hypothesis that intraoperative hypothermia prolongs postoperative recovery. METHODS: Patients undergoing elective major abdominal surgery (n = 150) were anesthetized with isoflurane, nitrous oxide, and fentanyl. They were randomly assigned to routine thermal management (hypothermia) or extra warming (normothermia). Postoperative surgical pain was treated with patient-controlled analgesia. Fitness for discharge from the postanesthesia care unit was evaluated at 20-min intervals by investigators blinded to group assignment and postoperative core temperatures. Scoring was based on a modification of a previously published system that included activity, ventilation, consciousness, and hemodynamic responses. Patients were considered fit for discharge when they sustained a score of 80% (13 points) for at least two consecutive measurement periods. RESULTS: Morphometric characteristics and anesthetic management were similar in each group. Final intraoperative core temperatures differed by approximately 2 degrees C: 34.8 +/- 0.6 versus 36.7 +/- 0.6 degrees C (mean +/- SD, P < 0.001). Postoperative pain scores and postoperative use of patient-controlled opioid were similar. Hypothermic patients required approximately 40 min longer (94 +/- 65 vs. 53 +/- 36 min) to reach fitness for discharge, even when return to normothermia was not a criterion (P < 0.001). Duration of recovery in the two groups differed by approximately 90 min when a core temperature >36 degrees C was also required (P < 0.001). CONCLUSION: Maintaining core normothermia decreases the duration of postanesthetic recovery and may, therefore, reduce costs of care. PMID- 9416716 TI - Oral clonidine premedication reduces minimum alveolar concentration of sevoflurane for tracheal intubation in children. AB - BACKGROUND: Sevoflurane is a useful anesthetic for inhalational induction in children because of its low solubility in blood and relatively nonpungent odor. Clonidine has sedative and anxiolytic properties and reduces the requirement for inhalation agents. Nitrous oxide (N2O) also decreases the requirement of inhaled anesthetics, but the effect is variable. The minimum alveolar concentration for tracheal intubation (MAC(TI)) of sevoflurane was assessed with and without N2O and clonidine premedication. METHODS: Seventy-two patients, aged 3-11 yr, were assigned to one of six groups (n = 12 each). They received one of three preanesthetic medications (two groups for each premedication): placebo (control), 2 microg/kg oral clonidine or 4 microg/kg oral clonidine. In one group of each premedication, anesthesia was induced with sevoflurane in oxygen; in the other group, anesthesia was induced with sevoflurane in the presence of 60% N2O. Each concentration of sevoflurane at which tracheal intubation was attempted was predetermined according to Dixon's up-and-down method and held constant for at least 20 min before the trial RESULTS: The MAC(TI) of sevoflurane in the absence of N2O (mean +/- SEM) was 3.2 +/- 0.2%, 2.5 +/- 0.1%, and 1.9 +/- 0.2% in the control, 2-microg/kg clonidine, and 4-microg/kg clonidine groups, respectively. Nitrous oxide (60%) decreased the MAC(TI) of sevoflurane by 26%, 24%, and 27% in the control, 2-microg/kg clonidine, and 4-microg/kg clonidine groups. CONCLUSIONS: Oral clonidine premedication decreased the MAC(TI) of sevoflurane. Nitrous oxide also decreased the MAC(TI). The combination of clonidine and N2O lessened the MAC(TI) of sevoflurane more than did either drug alone. PMID- 9416717 TI - Ketamine decreases intracranial pressure and electroencephalographic activity in traumatic brain injury patients during propofol sedation. AB - BACKGROUND: The potential adverse effects of ketamine in neurosurgical anesthesia have been well established and involve increased intracranial pressure (ICP) and cerebral blood flow. However, reexamination of ketamine is warranted because data regarding the effects of ketamine on cerebral hemodynamics are conflicting. METHODS: Eight patients with traumatic brain injury were studied. In all patients, ICP monitoring was instituted before the study. Control of ICP (less than 25 mmHg), hemodynamic values, and blood gas tension (partial pressure of carbon dioxide in arterial blood between 35-38 mmHg) was obtained with propofol infusion (3 mg x kg(-1) x h(-1)) and mechanical ventilation. The effects of three doses of ketamine, 1.5, 3, and 5 mg/kg, respectively, on ICP, cerebral perfusion pressure, jugular vein bulb oxygen saturation, middle cerebral artery blood flow velocity, and electric activity of the brain (EEG) were measured. The three doses were administered intravenously at 6-h intervals over 30 s through a central venous line. Systemic and cerebral hemodynamics and end-tidal carbon dioxide were continuously monitored and recorded at 1-min intervals throughout the 30-min study periods. RESULTS: Ketamine, in all three doses studied (1.5, 3, and 5 mg/kg) was associated with a significant decrease in ICP (mean +/- SD: 2 +/- 0.5 mmHg [P < 0.05], 4 +/- 1 mmHg [P < 0.05], and 5 +/- 2 mmHg [P < 0.05]) among the study patients regardless of the ketamine dose used. There were no significant differences in cerebral perfusion pressure, jugular vein bulb oxygen saturation, and middle cerebral artery blood flow velocity. Ketamine induced a low-amplitude fast-activity electroencephalogram, with marked depression, such as burst suppression. CONCLUSIONS: These results suggest that ketamine may not adversely alter cerebral hemodynamics of mechanically ventilated head-trauma patients sedated with propofol. These encouraging results should be confirmed in larger groups of similar patients. PMID- 9416718 TI - Comparison of the Bullard and Macintosh laryngoscopes for endotracheal intubation of patients with a potential cervical spine injury. AB - BACKGROUND: In the emergency trauma situation, in-line stabilization (ILS) of the cervical spine is used to reduce head and neck extension during laryngoscopy. The Bullard laryngoscope may result in less cervical spine movement than the Macintosh laryngoscope. The aim of this study was to compare cervical spine extension (measured radiographically) and time to intubation with the Bullard and Macintosh laryngoscopes during a simulated emergency with cervical spine precautions taken. METHODS: Twenty-nine patients requiring general anesthesia and endotracheal intubation were studied. Patients were placed on a rigid board and anesthesia was induced. Laryngoscopy was performed on four occasions: with the Bullard and Macintosh laryngoscopes both with and without manual ILS. Cricoid pressure was applied with ILS. To determine cervical spine extension, radiographs were exposed before and during laryngoscopy. Times to intubation and grade view of the larynx were also compared. RESULTS: Cervical spine extension (occiput-C5) was greatest with the Macintosh laryngoscope (25.9 degrees +/- 2.8 degrees). Extension was reduced when using the Macintosh laryngoscope with ILS (12.9 +/- 2.1 degrees) and the Bullard laryngoscope without stabilization (12.6 +/- 1.8 degrees; P < 0.05). Times to intubation were similar for the Macintosh laryngoscope with ILS (20.3 +/- 12.8 s) and for the Bullard without ILS (25.6 +/- 10.4 s). Manual ILS with the Bullard laryngoscope results in further reduction in cervical spine extension (5.6 +/- 1.5 degrees) but prolongs time to intubation (40.3 +/- 19.5 s; P < 0.05). CONCLUSIONS: Cervical spine extension and time to intubation are similar for the Macintosh laryngoscope with ILS and the Bullard laryngoscope without ILS. However, time to intubation is significantly prolonged when the Bullard laryngoscope is used in a simulated emergency with cervical spine precautions taken. This suggests that the Bullard laryngoscope may be a useful adjunct to intubation of patients with potential cervical spine injury when time to intubation is not critical. PMID- 9416719 TI - Patient-controlled analgesia after major shoulder surgery: patient-controlled interscalene analgesia versus patient-controlled analgesia. AB - BACKGROUND: The authors compared patient-controlled interscalene analgesia (PCIA) with local anesthetics with intravenous patient-controlled analgesia (PCA) with opioids to manage postoperative pain after major shoulder surgery. METHODS: Forty patients scheduled for elective major shoulder surgery were prospectively randomized to receive either PCIA or PCA. Before surgery, all patients had an interscalene block. In the PCIA group, a catheter was introduced within the interscalene sheath. Six hours after the initial block, patients received for 48 h either a continuous infusion of 0.15% bupivacaine through the interscalene catheter at a rate of 5 ml/h plus a bolus of 3 or 4 ml with a lock-time of 20 min (group PCLA) or a continuous intravenous infusion of nicomorphine at a rate of 0.5 mg/h plus a bolus of 2 or 3 mg with a lock-time of 20 min (group PCA). Pain relief was regularly assessed using a visual analog scale, side effects were noted, and the patients were asked to rate their satisfaction at the end of the study. RESULTS: Pain relief was significantly better controlled in the PCIA group at t = 12 and 18 h (P < 0.05). Vomiting and pruritus were 0 versus 25% and 0 versus 25% for the PCIA and PCA groups, respectively (P < 0.05). Patient satisfaction was greater in the PCIA group (P < 0.05). Time of first bolus administration and paracetamol supplement were similar in both groups. CONCLUSIONS: The use of the PCIA technique was uncomplicated and provided better pain relief than PCA during the first 18 h after operation. The incidence of side effects such as vomiting and pruritus was significantly decreased with the use of PCIA, and patient satisfaction was superior in the PCIA group. PMID- 9416720 TI - Lack of analgesic activity of morphine-6-glucuronide after short-term intravenous administration in healthy volunteers. AB - BACKGROUND: The analgesic activity of morphine-6-glucuronide (M-6-G) is well recognized for its contribution to the effects of morphine and its possible use as an opioid analgesic with a wider therapeutic range than morphine. The present study attempted to quantify the relative contribution of M-6-G to analgesia observed after systemic administration of morphine. METHODS: In a placebo controlled, sixfold crossover study in 20 healthy men, the effects of M-6-G were assessed at steady-state plasma concentrations of M-6-G identical to and two and three times higher than those measured after administration of morphine. Morphine and M-6-G were administered as an intravenous bolus followed by infusion over 4 h. Dosage A was M-6-G-bolus of 0.015 mg/kg plus infusion of 0.0072 mg x kg(-1) x h(-1). Dosage B was M-6-G-bolus of 0.029 mg/kg plus infusion of 0.014 mg x kg(-1) x h(-1). Dosage C was M-6-G-bolus of 0.044 mg/kg plus infusion of 0.022 mg x kg( 1) x h(-1). Dosage D was a morphine bolus of 0.14 mg/kg plus infusion of 0.05 mg x kg(-1) x h(-1) for 4 h. Dosage E was M-6-G combined with morphine (doses A + D). Dosage F was a placebo. The analgesic effects of M-6-G and morphine were measured before administration of the bolus and after 3.5 h using an experimental pain model based on pain-related cortical potentials and pain ratings after specific stimulation of the nasal nociceptor with short pulses of gaseous carbon dioxide. RESULTS: Morphine significantly reduced subjective and objective pain correlates compared with placebo. In contrast, M-6-G produced no statistically significant effects. The addition of M-6-G to morphine did not increase the effects of morphine. Morphine produced significantly more side effects than M-6 G. CONCLUSION: After short-term intravenous administration at doses that produce plasma concentrations of M-6-G similar to those seen after administration of morphine, M-6-G had no analgesic effects in the present placebo-controlled study in healthy volunteers. PMID- 9416721 TI - Sympathovagal effects of spinal anesthesia assessed by the spontaneous cardiac baroreflex. AB - BACKGROUND: The changes in sympathovagal balance induced by spinal anesthesia remain controversial. The spontaneous baroreflex method allows the continuous assessment of the spontaneous engagement of the cardiac baroreflex, giving an index of sympathovagal balance. The purpose of this study was to follow the effects of spinal anesthesia on spontaneous baroreflex sensitivity. METHODS: Continuous electrocardiogram and noninvasive blood pressure were recorded in 24 patients scheduled for elective inguinal hernia repair and randomly assigned to three groups: (1) no volume loading, (2) volume loading of 15 ml/kg lactated Ringer's solution, and (3) continuous infusion of etilefrine (an ephedrine-like drug). Each patient was studied before, during, and after bupivacaine-induced spinal anesthesia (mean sensory block: T4). Spontaneous baroreflex sensitivity and parameters of time-domain analysis of heart rate variability were calculated from 30 min of recording of each period. RESULTS: No significant change in spontaneous baroreflex slope or parameters of time-domain analysis were observed after regional anesthesia in any group. However, three patients experienced episodes of bradycardia and hypotension in the absence of a high block; these three patients showed an increase in spontaneous baroreflex sensitivity and time domain parameters. CONCLUSIONS: Using a noninvasive, continuous technique to estimate cardiac sympathovagal balance, no significant variation in autonomic balance induced by spinal anesthesia was observed. However, untoward episodes of bradycardia and hypotension occurred in three patients, who could not be prospectively identified by the parameters studied. PMID- 9416722 TI - Dose-response of rocuronium bromide in children anesthetized with propofol: a comparison with succinylcholine. AB - BACKGROUND: The aim of this study was to determine the potency of rocuronium during propofol/fentanyl/N2O anesthesia in children and to compare the time course of action of rocuronium at doses of two and three times the ED95 with that of succinylcholine. METHODS: Rocuronium (120, 160, 200, or 240 microg/kg) was administered to 48 children aged 2-10 yr. Neuromuscular block was assessed by monitoring the electromyographic response of the adductor digiti minimi to supramaximal stimulation of the ulnar nerve at 2 Hz for 2 s every 10 s. Potency was determined by log-probit transformation and least-squares linear regression analysis of dose and response. In a second group of 30 children, the onset and recovery profile of rocuronium at doses of two and three times the ED95 was compared with that of succinylcholine (2 mg/kg). RESULTS: Values for ED50 and ED95 were 210 +/- 24 and 404 +/- 135 microg/kg, respectively. The time to 90% neuromuscular block after 1.2 mg/kg rocuronium (three times the ED95), 33 +/- 5 s (mean +/- SD), did not differ significantly from that after succinylcholine, at 30 +/- 7 s; however, both were significantly less than that after 0.8 mg/kg rocuronium, 46 +/- 8 s (P < 0.05). The time to 25% recovery from 1.2 microg/kg rocuronium, 41 +/- 13 min, was approximately 50% greater than that after 0.8 mg/kg, at 27 +/- 6 min (P < 0.001), and eight times greater than that after succinylcholine, at 5.2 +/- 1.9 min (P < 0.001). CONCLUSIONS: Both 1.2 mg/kg rocuronium (three times the ED95) and 2 mg/kg succinylcholine provide 90% neuromuscular block within 45 s in 95% of children. The present dose-response data support the use of rocuronium at a dose of 1.2 mg/kg when rapid onset and intermediate-duration neuromuscular block are needed in children. PMID- 9416723 TI - Cost-benefit and efficacy of aprotinin compared with epsilon-aminocaproic acid in patients having repeated cardiac operations: a randomized, blinded clinical trial. AB - BACKGROUND: Aprotinin and epsilon-aminocaproic acid are routinely used to reduce bleeding during cardiac surgery. The marked difference in average wholesale cost between these two drug therapies (aprotinin, $1,080 vs. epsilon-aminocaproic acid, $11) has generated significant controversy regarding their relative efficacies and costs. METHODS: In a multicenter, randomized, prospective, blinded trial, patients having repeated cardiac surgery received either a high-dose regimen of aprotinin (total dose, 6 x 10(6) kallikrein inactivator units) or epsilon-aminocaproic acid (total dose, 270 mg/kg). RESULTS: Two hundred four patients were studied. Overall (data are median [25th-75th percentiles]), aprotinin-treated patients had less postoperative thoracic drainage (511 ml [383 805 ml] vs. 655 ml [464-1,045 ml]; P = 0.016) and received fewer platelet transfusions (0 [range, 0-1] vs. 1 [range, 0-2]; P = 0.036). The surgical field was more likely to be considered free of bleeding in aprotinin-treated patients (44% vs. 26%; P = 0.012). No differences, however, were seen in allogeneic erythrocyte transfusions or in the time required for chest closure. Overall, direct and indirect bleeding-related costs were greater in aprotinin- than in epsilon-aminocaproic acid-treated patients ($1,813 [$1,476-2,605] vs. $1,088 [range, $511-2,057]; P = 0.0001). This difference in cost per case varied in magnitude among sites but not in direction. CONCLUSIONS: Aprotinin was more effective than epsilon-aminocaproic acid at decreasing bleeding and platelet transfusions. Epsilon-aminocaproic acid, however, was the more cost-effective therapy over a broad range of estimates for bleeding-related costs in patients undergoing repeated cardiac surgery. A cost-benefit analysis using the lower cost of half-dose aprotinin ($540) still resulted in a significant cost advantage using epsilon-aminocaproic therapy (P = 0.022). PMID- 9416724 TI - The effect of halothane on the recirculatory pharmacokinetics of physiologic markers. AB - BACKGROUND: The cardiovascular effects of halothane are well recognized, but little is known of how this affects drug distribution. The effect of halothane anesthesia on physiologic factors that affect drug disposition from the moment of injection was investigated. METHODS: The dispositions of markers of intravascular space and blood flow (indocyanine green), extracellular space and free water diffusion (inulin), and total body water and tissue perfusion (antipyrine) were determined in four purpose-bred coonhounds. The dogs were studied while awake and while anesthetized with 1%, 1.5%, and 2% halothane in a randomized order determined by a repeated measures Latin square experimental design. Marker dispositions were described by recirculatory pharmacokinetic models based on frequent early and less frequent later arterial blood samples. These models characterize the role of cardiac output and its distribution on drug disposition. RESULTS: Halothane caused a significant and dose-dependent decrease in cardiac output. The disposition of antipyrine was most profoundly affected by halothane anesthesia, which increased both nondistributive intercompartmental clearance and volume while decreasing fast and slow tissue clearances and elimination clearance in a halothane dose-dependent manner. CONCLUSIONS: Halothane-induced changes in blood flow to the compartments of the antipyrine recirculatory model were not proportional to changes in cardiac output. Halothane anesthesia significantly increased (to more than double) the area under the drug concentration versus time curve due to an increase in the apparent peripheral blood flow not involved in drug distribution, despite a dose-dependent cardiac output decrease. Recirculatory pharmacokinetic models include the best aspects of traditional compartmental and physiologic pharmacokinetic models while offering advantages over both. PMID- 9416725 TI - Emulsion formulation reduces propofol's dose requirements and enhances safety. AB - BACKGROUND: Propofol, a highly lipophilic anesthetic, is formulated in a lipid emulsion for intravenous use. Propofol has brisk onset and offset of effect after rapid administration and retains rapid offset characteristics after long-term administration. The authors tried to determine whether the emulsion vehicle is requisite for propofol's evanescent effect-time profile. METHODS: The time course of sedation and electroencephalographic (EEG) effect after propofol administration was measured in three studies in rats instrumented. In study 1, propofol was infused in either emulsion or lipid-free vehicle (n = 12), in a repeated measures cross-over design. In study 2, propofol in lipid-free vehicle was infused with or without simultaneous infusion of drug-free lipid emulsion (n = 6) in a repeated measures cross-over design. In study 3, propofol was infused in either emulsion (n = 5) or lipid-free vehicle (n = 5) to EEG burst suppression. RESULTS: In study 1, relative to the emulsion formulation, propofol administered at equivalent doses in lipid-free vehicle resulted in a longer time to effect onset (1.4 +/- 0.2 vs. 0.5 +/- 0.1 min, EEG) and a trend for delayed anesthetic recovery (26.8 +/- 9.4 vs. 17 +/- 3.5 min, EEG; 26.1 +/- 8.8 vs. 16.8 +/- 3.3 min, sleep). In study 2, coadministration of drug-free emulsion with propofol did not alter the time course of effect. In study 3, more than twice the dose of propofol was required to achieve EEG burst suppression with the lipid free formulation. Two animals died after administration of propofol to EEG burst suppression with the lipid-free formulation; no deaths occurred in the emulsion group. CONCLUSION: The incorporation of propofol in emulsion reduces dose requirements and produces rapid onset and recovery of anesthetic effect. PMID- 9416726 TI - Myocardial effects of halothane and isoflurane in hamsters with hypertrophic cardiomyopathy. AB - BACKGROUND: The effects of halothane and isoflurane on myocardial contraction and relaxation in diseased myocardium are not completely understood. METHODS: The effects of equianesthetic concentrations of halothane and isoflurane on inotropy and lusitropy in left ventricular papillary muscles of healthy hamsters and those with genetically induced cardiomyopathy (strain BIO 14.6) were investigated in vitro (29 degrees C; pH 7.40; Ca2+ 2.5 mM; stimulation frequency, 3/min) in isotonic and isometric conditions. RESULTS: Halothane induced a negative inotropic effect that was greater in cardiomyopathic than in healthy hamsters (1.5 vol%, active isometric force (AF): 19 +/- 8% vs. 28 +/- 11% of control values; P < 0.05). Isoflurane induced a negative inotropic effect that was greater in cardiomyopathic than in healthy hamsters (2.0 vol%, AF: 64 +/- 13% vs. 75 +/- 11% of control values; P < 0.01). However, the negative inotropic effects of halothane and isoflurane were not different for cardiomyopathic or healthy hamsters when their concentrations were corrected for minimum alveolar concentration (MAC) values in each strain. Halothane induced a negative lusitropic effect under low load, which was more important in cardiomyopathic hamsters, suggesting a greater impairment in calcium uptake by the sarcoplasmic reticulum. In contrast, isoflurane induced a moderate positive lusitropic effect under low load in healthy but not in cardiomyopathic hamsters. Halothane and isoflurane induced no significant lusitropic effect under high load. CONCLUSIONS: Halothane and isoflurane had greater negative inotropic effects in cardiomyopathic than in healthy hamsters. Nevertheless, no significant differences in their inotropic effects were noted when concentrations were correlated as a multiple of MAC in each strain. PMID- 9416727 TI - Effects of ketamine on ventricular conduction, refractoriness, and wavelength: potential antiarrhythmic effects: a high-resolution epicardial mapping in rabbit hearts. AB - BACKGROUND: The aims of the study were to verify the effects of ketamine on ventricular conduction velocity and on the ventricular effective refractory period, to determine its effects on anisotropy and on homogeneity of refractoriness, and to use wavelength to determine whether ketamine has antiarrhythmic or arrhythmogenic properties. METHODS: A high-resolution epicardial mapping system was used to study the effects of 50, 100, 150, and 200 microM racemic ketamine in 15 isolated, Langendorff-perfused rabbit hearts. Five hearts were kept intact to study the effects of ketamine on spontaneous sinus cycle length (RR) interval and its putative arrhythmogenic effects. In 10 other hearts, a thin epicardial layer was obtained by an endocardial cryoprocedure (frozen hearts) to study ventricular conduction velocity, ventricular effective refractory periods (five sites), and ventricular wavelength. RESULTS: Ketamine induced a concentration-dependent lengthening of the RR interval. Ketamine slowed longitudinal and transverse ventricular conduction velocity with no anisotropic change, and it prolonged the ventricular effective refractory period with no significant increase in dispersion. Ventricular longitudinal and transverse wavelengths tend to increase, but this was not statistically significant. Finally, no arrhythmia could be induced regardless of the ketamine concentration. CONCLUSION: Ketamine slowed ventricular conduction and prolonged refractoriness without changing anisotropy or increasing dispersion of refractoriness. Although these effects should result in significant antiarrhythmic effects of ketamine, this should not be construed to suggest a protective effect in ischemic or other abnormal myocardium. PMID- 9416728 TI - Dose-dependent effects of halothane on the phrenic nerve responses to acute hypoxia in vagotomized dogs. AB - BACKGROUND: Previous studies in dogs and humans suggest that the carotid body chemoreceptor response to hypoxia is selectively impaired by halothane. The present studies in an open-loop canine preparation were performed to better delineate the effects of anesthetic concentrations of halothane on the carotid body chemoreceptor-mediated phrenic nerve response to an acute hypoxic stimulus. METHODS: Three protocols were performed to study the effects of halothane anesthesia on the phrenic nerve response to 1 min of isocapnic hypoxia (partial pressure of oxygen [PaO2] at peak hypoxia, 35-38 mmHg) in unpremedicated, anesthetized, paralyzed, vagotomized dogs during constant mechanical ventilation. In protocol 1, the dose-dependent effects of halothane from 0.5-2.0 minimum alveolar concentration (MAC) on the hypoxic response during moderate hypercapnia (partial pressure of carbon dioxide [PaCO2], 60-65 mmHg) were studied in 10 animals. In protocol 2, the hypoxic responses at 1 MAC halothane near normocapnia (PaCO2, 40-45 mmHg) and during moderate hypercapnia were compared in an additional four animals. In protocol 3, the hypoxic response of 4 of 10 dogs from protocol 1 was also studied under sodium thiopental (STP) anesthesia after they completed protocol 1. RESULTS: Protocol 1: Peak phrenic nerve activity (PPA) increased significantly during the hypoxic runs compared with the isocapnic hyperoxic controls at all halothane doses. The phrenic nerve response to the hypoxic stimulus was present even at the 2 MAC dose. Protocol 2: The net hypoxic responses for the two carbon dioxide background levels at 1 MAC were not significantly different. Protocol 3: The net hypoxic response of PPA for the STP anesthetic was not significantly different from the 1 MAC halothane dose. Bilateral carotid sinus denervation abolished the PPA response to hypoxia. CONCLUSIONS: The phrenic nerve response to an acute, moderately severe isocapnic hypoxic stimulus is dose-dependently depressed but not abolished by surgical doses of halothane. This analysis does not suggest a selective depression of the carotid body chemoreceptor response by halothane. The observed hypoxic phrenic response was mediated by the carotid body chemoreceptors in vagotomized dogs because bilateral carotid sinus denervation abolished all increases in PPA. PMID- 9416729 TI - Effects of halothane on the phrenic nerve responses to carbon dioxide mediated by carotid body chemoreceptors in vagotomized dogs. AB - BACKGROUND: Previous studies in dogs showed that the phrenic nerve response to an acute hypoxic stimulus was dose dependently depressed by 0.5-2.0 minimum alveolar concentration (MAC) of halothane but not abolished. Because a carbon dioxide stimulus is transduced by a different mechanism in the carotid body chemoreceptors (CBCRs) than is a hypoxic stimulus, inhalational anesthetics may preferentially depress one of these transduction processes, the central neuronal processing, or both, of the integrated responses to these two types of inputs. METHODS: Carotid body chemoreceptor stimulation was produced by short (1-1.5 s), bilateral, 100% carbon dioxide in saline infusions into the carotid arteries during neural inspiration in unpremedicated, halothane-anesthetized, paralyzed, vagotomized dogs during constant mechanical ventilation. The phrenic neurogram quantified the neural inspiratory response. Four protocols were performed in the study: (1) the dose-dependent effects of halothane anesthesia (0.5-2.0 MAC) during hyperoxic hypercapnia on phrenic nerve activity, (2) the effects of three background levels of the partial pressure of carbon dioxide (PaCO2) on the magnitude of the carbon dioxide infusion responses at 1 MAC halothane, (3) the effects of anesthetic type on the magnitude of the carbon dioxide infusion response, and (4) the effects of CBCR denervation. RESULTS: Peak phrenic nerve activity (PPA) increased significantly during the carbon dioxide-stimulated phrenic burst in protocols 1-3; after denervation there was no response (protocol 4). Halothane produced a dose-dependent reduction in the PPA of control and carbon dioxide infusion-stimulated phrenic bursts and in the net carbon dioxide response. The net PPA responses for the different PaCO2 background levels were not different but were somewhat larger for sodium thiopental anesthesia than for 1.0 MAC halothane. CONCLUSIONS: The phrenic nerve response to an acute, severe carbon dioxide stimulus was dose dependently depressed by surgical doses of halothane. The observed responses to carbon dioxide infusion were mediated by the CBCRs because they were eliminated by CBCR denervation. These results suggest that the CBCR transduction and central transmission of the carbon dioxide signal in terms of inspiratory excitatory drive are not abolished at surgical levels of halothane anesthesia. PMID- 9416730 TI - Involvement of cerulospinal glutamatergic neurotransmission in fentanyl-induced muscular rigidity in the rat. AB - BACKGROUND: Investigators in the authors' laboratory previously established the critical participation of the cerulospinal noradrenergic pathway in muscular rigidity elicited by fentanyl. The identification of colocalization of glutamate with tyrosine hydroxylase in most locus ceruleus neurons suggests a role for cerulospinal glutamatergic neurotransmission in fentanyl-induced muscular rigidity. This suggestion and the subtype(s) of glutamate receptors involved were investigated here. METHODS: Electromyographic signals activated by bilateral microinjection of 2.5 microg fentanyl into the locus ceruleus were recorded differentially from the left sacrococcygeus dorsi lateralis muscle of adult male Sprague-Dawley rats. The effect of intrathecal administration at the lower lumbar spinal cord of various N-methyl-D-aspartate (NMDA) and non-NMDA receptor antagonists or agonists on this index of muscular rigidity was studied. Rats were under mechanical ventilation, and intravenous infusion of ketamine (30 mg x kg( 1) x h(-1)) was maintained until 10 min before fentanyl was administered. RESULTS: Microinjection of fentanyl bilaterally into the locus ceruleus increased the root mean square and decreased the mean power frequency values of electromyographic signals. The efficacy of fentanyl to elicit muscular rigidity in this manner was significantly reduced by previous intrathecal administration of either 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), (+)-5-methyl-10,11-dihydro 5H-dibenzo[a,d]cyclohepten-5,10-imine maleate (MK-801), D-(-)-2-amino-5 phosphonovaleric acid (AP5), or (+/-)-3-(2-carboxypiperazin-4-yl)-propyl-1 phosphonic acid (CPP). Intrathecal administration of kainic acid or NMDA also resulted in significant electromyographic activation. CONCLUSIONS: In addition to the cerulospinal noradrenergic mechanism, the cerulospinal glutamatergic pathway and both NMDA and non-NMDA receptors in the spinal cord may mediate fentanyl induced muscular rigidity in the rat. PMID- 9416731 TI - Interaction between anesthetics and the sodium-hydrogen exchange inhibitor HOE 642 (cariporide) in ischemic and reperfused rat hearts. AB - BACKGROUND: Sodium (Na+)-hydrogen (H+) exchange (NHE) inhibitors are effective cardioprotective agents. The potent NHE inhibitor HOE 642 (cariporide) is being evaluated clinically in high-risk patients, including those having coronary artery bypass. Volatile anesthetics are also cardioprotective, most likely via different mechanisms. The potential interaction between anesthetics and HOE 642 was investigated. METHODS: Electrically paced isolated rat hearts were perfused at constant flow. Left ventricular developed pressure and end-diastolic pressure were monitored as determinants of function. Hearts were subjected to 60 min each of total ischemia followed by reperfusion. Isoflurane (0.93 minimum alveolar concentration [MAC]), sevoflurane (1.03 MAC), or sufentanil (1.2 nM) was added 15 min before ischemia and throughout reperfusion, either alone or in combination with HOE 642 (5 microM). The effect of HOE 642 alone was also studied. At the end of reperfusion, hearts were freeze-clamped for subsequent determination of tissue metabolites. RESULTS: In control hearts, left ventricular developed pressure recovered to 40% of preischemia values, whereas left ventricular end-diastolic pressure increased by 650% after reperfusion. Sevoflurane, isoflurane, or HOE 642 alone significantly enhanced left ventricular developed pressure recovery to more than 90%, although recovery with HOE 642 was more rapid and accompanied by significantly reduced left ventricular end-diastolic pressure. HOE 642 plus volatile anesthetics produced additive effects, with left ventricular developed pressure recovering by more that 100%, although left ventricular end-diastolic pressure was not further reduced. Sufentanil had no effect in terms of developed pressure, but protection with HOE 642 was maintained. HOE 642 with or without volatile anesthetics also preserved adenosine triphosphate levels. CONCLUSIONS: Isoflurane, sevoflurane, and HOE 642 enhance ventricular recovery, but the effect of HOE 642 is also associated with reduced contracture and adenosine triphosphate preservation. A combination of the NHE inhibitor and either volatile agent confers additive and superior protection, which could be relevant for the establishment of ideal cardioprotective strategies during surgery. PMID- 9416732 TI - Lidocaine suppresses the anoxic depolarization and reduces the increase in the intracellular Ca2+ concentration in gerbil hippocampal neurons. AB - BACKGROUND: The movement of ions, particularly Ca2+, across the plasma membrane of neurons is regarded as an initial element of the development of ischemic neuronal damage. Because the mechanism by which lidocaine protects neurons against ischemia is unclear, the effects of lidocaine on the ischemia-induced membrane depolarization, histologic outcome, and the change in the intracellular Ca2+ concentration in the gerbil hippocampus were studied. METHODS: The changes in the direct-current potential shift in the hippocampal CA1 area produced by transient forebrain ischemia for 4 min were compared in animals given lidocaine (0.8 micromol administered intracerebroventricularly) 10 min before ischemia and those given saline. The histologic outcome was evaluated 7 days after ischemia by assessing delayed neuronal death in hippocampal CA1 pyramidal cells in these animals. In a second study, hypoxia-induced intracellular Ca2+ increases were evaluated by in vitro microfluorometry in gerbil hippocampal slices, and the effects of lidocaine (10, 50, and 100 microM) on the Ca2+ accumulation were examined. In addition, the effect of lidocaine (100 microM) drug perfusion with a Ca2+-free ischemia-like medium was investigated. RESULTS: The preischemic administration of lidocaine delayed the onset of the ischemia-induced membrane depolarization (anoxic depolarization) and reduced its maximal amplitude. The histologic outcome was improved by the preischemic treatment with lidocaine. The in vitro hypoxia-induced increase in the intracellular concentration of Ca2+ was suppressed by the perfusion with lidocaine-containing mediums (50 and 100 microM), regarding the initiation and the extent of the increase. The hypoxia induced intracellular Ca2+ elevation in the Ca2+-free condition was similar to that in the Ca2+-containing condition. Perfusion with lidocaine (100 microM) inhibited this elevation in the Ca2+-free condition. CONCLUSIONS: Lidocaine helps protect neurons from ischemia by suppressing the direct-current potential shift, by inhibiting the release of Ca2+ from the intracellular Ca2+ stores, and by inhibiting the influx from the extracellular space. PMID- 9416733 TI - Inhibition of nitric oxide synthase decreases anesthetic requirements of intravenous anesthetics in Xenopus laevis. AB - BACKGROUND: Acute inhibition of nitric oxide synthase (NOS) has been demonstrated to reduce the anesthetic requirements of volatile anesthetics. Recent data suggest that not only volatile but also intravenous anesthetic agents interact with nitric oxide (NO) metabolism. The aim of this study was to examine the effect of NOS inhibition by nitroG-L-arginine-methyl-ester (L-NAME) on the anesthetic action of the intravenous anesthetics thiopental, propofol, and ketamine. METHODS: The anesthetic potencies of thiopental, propofol, and ketamine were determined in Xenopus laevis tadpoles in the absence and presence of L-NAME. Anesthesia was defined as loss of righting reflex for 5 s. A nonlinear logistic regression curve was fitted to the data and half-maximal effective concentrations (EC50) were calculated. A second set of experiments was performed with different concentrations of L-NAME in the presence of the previously determined the EC50 of the intravenous anesthetics. RESULTS: The EC50s of the anesthetics thiopental, propofol, and ketamine were determined to be 25.5 +/- 2.0 microM, 1.9 +/- 0.1 microM, and 59.7 +/- 0.7 microM, respectively. The addition of L-NAME shifted the concentration-response curves to the left in a concentration-dependent manner. In the presence of 1 mM L-NAME, the EC50 of thiopental was reduced by 43%, the EC50 of propofol by 26%, and the EC50 of ketamine by 63%. The addition of D-NAME did not change the EC50 values of the three anesthetics. In the presence of L arginine, the effect of L-NAME on the EC50 of thiopental was reversed. When administered by itself in a concentration range from 0.1 microM to 10 mM, L-NAME did not alter the behavior of the tadpoles. CONCLUSIONS: The results of the present study show that acute inhibition of NOS by L-NAME results in reduced anesthetic requirements of the intravenous anesthetics thiopental, propofol, and ketamine. This interaction of acutely administered L-NAME and intravenous anesthetics indicates that the NO-cyclic guanosine 3',5'-monophosphate system is involved in mediating the anesthetic effect of these compounds. PMID- 9416734 TI - Experimental subarachnoid hemorrhage in rats: effect of intravenous alpha-alpha diaspirin crosslinked hemoglobin on hypoperfusion and neuronal death. AB - BACKGROUND: Hemodilution with diaspirin crosslinked hemoglobin (DCLHb) ameliorates occlusive cerebral ischemia. However, subarachnoid hemoglobin has been implicated as a cause of cerebral hypoperfusion. The effect of intravenous DCLHb on cerebral perfusion and neuronal death after experimental subarachnoid hemorrhage was evaluated. METHODS: Rats (n = 48) were anesthetized with isoflurane and subarachnoid hemorrhage was induced by injecting 0.3 ml of autologous blood into the cistema magna. Each animal received one of the following regimens: Control, no hematocrit manipulation; DCLHb, hematocrit concentration decreased to 30% with DCLHb; or Alb, hematocrit concentration decreased to 30% with human serum albumin. The experiments had two parts, A and B. In part A, after 20 min, cerebral blood flow (CBF) was assessed with 14C iodoantipyrine autoradiography. In part B, after 96 h, in separate animals, the number of dead neurons was determined in predetermined coronal sections by hematoxylin and eosin staining. RESULTS: Cerebral blood flow was greater for the DCLHb group than for the control group; and CBF was greater for the Alb group than the other two groups (P < 0.05). In one section, CBF was 45.5 +/- 10.9 ml x 100 g(-1) x min(-1) (mean +/- SD) for the control group, 95.3 +/- 16.6 ml x 100 g(-1) x min(-1) for the DCLHb group, and 138.1 +/- 18.7 ml x 100 g(-1) x min(-1) for the Alb group. The number of dead neurons was less in the Alb group (611 +/- 84) than in the control group (1,097 +/- 211), and was less in the DCLHb group (305 +/- 38) than in the other two groups (P < 0.05). CONCLUSIONS: These data support a hypothesis that hemodilution decreases hypoperfusion and neuronal death after subarachnoid hemorrhage. The data do not support the notion that intravascular molecular hemoglobin has an adverse effect on brain injury after subarachnoid hemorrhage. PMID- 9416735 TI - Conformational state-dependent effects of halothane on cardiac Na+ current. AB - BACKGROUND: The Na+ channel is voltage gated and characterized by three distinct states: closed, open, and inactivated. To identify the effects of halothane on the cardiac Na+ current (I(Na)) at various membrane potentials, the effects of 1.2 mM halothane at different holding potentials (V(H)) on I(Na) were examined in single, enzymatically isolated guinea pig ventricular myocytes. METHODS: The I(Na) was recorded using the whole-cell configuration of the patch-clamp technique. Currents were generated from resting V(H)s of -110, -80, or -65 mV. State-dependent block was characterized by monitoring frequency dependence, tonic block, and removal of inactivation by veratridine. RESULTS: Halothane produced significant (P < 0.05) V(H)-dependent depressions of peak I(Na) (mean +/- SEM): 24.4 +/- 4.1% (V(H) = -110 mV), 42.1 +/- 3.4% (V(H) = -80 mV), and 75.2 +/- 1.5% (V(H) = -65 mV). Recovery from inactivation was significantly increased when cells were held at -80 mV (control, tau = 6.0 +/- 0.3 ms; halothane, tau = 7.1 +/ 0.4 ms), but not at -110 mV. When using a V(H) of -80 mV, halothane exhibited a use-dependent block, with block of I(Na) increasing from 8.6 +/- 1.4% to 30.7 +/- 3.5% at test pulse rates of 2 and 11 Hz, respectively. Use-dependent inhibition was not apparent at V(H) of -110 mV. When inactivation of I(Na) was removed by exposure to 100 microM veratridine, no significant difference was observed in the depressant effect of halothane at both V(H)s: 26.6 +/- 4.5% (V(H) = -80 mV) and 26.4 +/- 5.6% (V(H) = -110 mV). CONCLUSIONS: The present findings indicate that the depressant action of halothane on cardiac I(Na) depends on the conformational state of the channel. As more channels are in the inactivated state, the more potent is the effect of halothane. Removal of channel inactivation by veratridine abolished the dependence of the halothane effect on V(H), but depression of the current was still evident. These results indicate a complex interaction between halothane and the various conformational states of the Na+ channel. PMID- 9416736 TI - Modulation of cardiac sodium current by alpha1-stimulation and volatile anesthetics. AB - BACKGROUND: Alpha1-adrenoceptor stimulation is known to produce electrophysiologic changes in cardiac tissues, which may involve modulations of the fast inward Na+ current (I(Na)). A direct prodysrhythmic alpha1-mediated interaction between catecholamines and halothane has been demonstrated, supporting the hypothesis that generation of halothane-epinephrine dysrhythmias may involve slowed conduction, leading to reentry. In this study, we examined the effects of a selective alpha1-adrenergic receptor agonist, methoxamine, on cardiac I(Na) in the absence and presence of equianesthetic concentrations of halothane and isoflurane in single ventricular myocytes from adult guinea pig hearts. METHODS: I(Na) was recorded using the standard whole-cell configuration of the patch-clamp technique. Voltage clamp protocols initiated from two different holding potentials (V(H)) were applied to examine state-dependent effects of methoxamine in the presence of anesthetics. Steady state activation and inactivation and recovery from inactivation were characterized using standard protocols. RESULTS: Methoxamine decreased I(Na) in a concentration- and voltage dependent manner, being more potent at the depolarized V(H). Halothane and isoflurane interacted synergistically with methoxamine to suppress I(Na) near the physiologic cardiac resting potential of -80 mV. The effect of methoxamine with anesthetics appeared to be additive when using a V(H) of -110 mV, a potential where no Na+ channels are in the inactivated state. Methoxamine in the absence and presence of anesthetics significantly shifted the half maximal inactivation voltage in the hyperpolarizing direction but had no effect on steady-state activation. CONCLUSION: The present results show that methoxamine (alpha1 adrenergic stimulation) decreases cardiac Na+ current in a concentration- and voltage-dependent manner. Further, a form of synergistic interaction between methoxamine and inhalational anesthetics, halothane and isoflurane, was observed. This interaction appears to depend on the fraction of Na+ channels in the inactivated state. (Key words: Anesthetics, volatile: halothane; isoflurane; methoxamine. Patch clamp: whole-cell configuration; sodium current; ventricular guinea pig myocytes.) PMID- 9416737 TI - Comparison of pre- versus post-incision administration of intrathecal bupivacaine and intrathecal morphine in a rat model of postoperative pain. AB - BACKGROUND: Preclinical studies in experimental animals suggest that preemptive analgesia may improve postoperative pain management. The beneficial effects of preemptive analgesia appear less remarkable clinically. The purpose of this study is to examine the effect of pre- and post-incision administration of intrathecal bupivacaine and intrathecal morphine in a rat model for postoperative pain. METHODS: Rats with intrathecal catheters were anesthetized with halothane, and the surgical field was prepared. A saline vehicle or the test drug was administered 15 min before an incision was made in the plantar aspect of the hindpaw or after the incision was completed. After recovery, mechanical hyperalgesia to punctate and nonpunctate stimuli was measured. Rats were tested on the day of surgery for the first 5 h and each day for 6 days. RESULTS: In saline vehicle-treated rats, the median withdrawal threshold decreased from 522 mN to 54 mN or less, and the response frequency to pressure from application of the plastic disc increased from 0 +/- 0% to 96 +/- 12% or greater after incision. Hyperalgesia was persistent through 2 days after surgery and then gradually returned toward preincision values over the next 4 days. Pre- or postincision administration of either intrathecal morphine or intrathecal bupivacaine reduced hyperalgesia on the day of surgery; at all subsequent times, there were no differences between the saline vehicle groups and the drug treatment groups. There were never any significant differences between pre- and postincision treatments. CONCLUSIONS: Early reduction in pain behaviors either by pre- or postincision management had no impact on subsequent measures of hyperalgesia in this model. These results agree with a number of clinical studies and suggest that incisional pain may be initiated and maintained differently than pain in other models. PMID- 9416738 TI - The immature heart and anesthesia. PMID- 9416740 TI - Use of a short flexible fiberoptic endoscope for difficult intubations. PMID- 9416739 TI - Propofol anesthesia and rational opioid selection: determination of optimal EC50 EC95 propofol-opioid concentrations that assure adequate anesthesia and a rapid return of consciousness. PMID- 9416741 TI - Susceptibility to malignant hyperthermia manifested as delayed return of increased serum creatine kinase activity and episodic rhabdomyolysis after exercise. PMID- 9416742 TI - Radicular pain due to a retained fragment of epidural catheter. PMID- 9416743 TI - Dysphagia with intrathecal fentanyl. PMID- 9416744 TI - One-lung ventilation for thoracotomy using a Hunsaker jet ventilation tube. PMID- 9416745 TI - Radiographic investigation of unilateral epidural block after single injection. PMID- 9416746 TI - Enoxaparin associated with psoas hematoma and lumbar plexopathy after lumbar plexus block. PMID- 9416748 TI - The successful implementation of pharmaceutical practice guidelines? Far from convincing! PMID- 9416747 TI - Electroencephalographic evidence of seizure activity under deep sevoflurane anesthesia in a nonepileptic patient. PMID- 9416749 TI - Economic analysis of anesthetic drug use. PMID- 9416750 TI - Implementation of pharmaceutical practice guidelines. PMID- 9416751 TI - Why must the practice of anesthesiology change? Economics is not the only thing! PMID- 9416753 TI - Optimal rehydration of desiccated CO2 absorbents. PMID- 9416752 TI - Anesthesia: coach, business, or first class? PMID- 9416754 TI - Inhaled nitric oxide delivery systems. PMID- 9416755 TI - An algorithm for quantifying blood pressure lability. PMID- 9416756 TI - Indicators of recovery of neuromuscular function. PMID- 9416757 TI - Increase in the arterial-to-end-tidal gradient (intraabdominal carbon dioxide insufflation in the pregnant ewe) PMID- 9416758 TI - Arterial to end-tidal gradients in pregnant subjects. PMID- 9416759 TI - Lidocaine to topically anesthetize the mucosal lining of the airway. PMID- 9416760 TI - A potential circuit leak with Tec 5 vaporizers. PMID- 9416761 TI - Changes in structural and functional properties of rat intestinal brush border membrane during starvation. AB - Changes in surface area of microvilli, fluidity of brush border membrane and transport of L-amino acids through intestinal epithelial cells were studied in wellfed and starved (2,4 and 6 days) rats. The surface area of microvilli per unit area of intestinal epithelial cells increased during starvation. Studies with fluoroprobes - pyrene, 1-anilinonaphthalene-8-sulphonate and 1,6-diphenyl 1,3,5-hexatriene, showed increased fluidity of brush border membrane on progressive starvation. Transport of five amino acids representing five different transport systems was studied during starvation in everted intestinal sleeves. Transport of L-proline, glycine and L-glutamic acid which represent imino, glycine and acidic systems respectively increased significantly in Na+-dependent pathway whereas transport of L-lysine representing basic system increased significantly in Na+-independent pathway during starvation. PMID- 9416762 TI - Potential antidepressant properties of forskolin and a novel water-soluble forskolin (NKH477) in the forced swimming test. AB - The mechanisms of the antidepressant activity of forskolin and a novel water soluble forskolin analog (NKH477) were studied using the forced swimming method in rats. Forskolin (0.01-0.1 mg/kg) and NKH477 (0.01-0.1 mg/kg) dose-dependently decreased ratings of immobility, with effects similar to those of amitriptyline treatment. The maximum effects of forskolin and NKH477 were observed at 0.01 mg/kg dose which is 150 more times potent than that (15 mg/kg) of amitriptyline. At a high dose (1.0 mg/kg) of forskolin and NKH477, the duration of immobility was returned to control levels. Forskolin and NKH477 did not influence the spontaneous locomotor activity at intraperitoneal injection doses from 0.01 to 1 mg/kg. Furthermore chronic administration with NKH477 at oral dose from 0.5 to 1.5 mg/kg significantly decreases the duration of immobility. These data indicate that both forskolin and NKH477 have strong antidepressive potency, consistent with the hypothesis that elevation of the cAMP cascade system may have an important role in antidepressive effects. PMID- 9416763 TI - Pharmacological evidence for the contribution of polyamines as mediators of the transcriptional component involved in smooth muscle relaxation elicited by forskolin. AB - To study whether cAMP-dependent transcriptional effect and polyamines might play a modulatory role on smooth muscle, the effect of forskolin on KCl (60 mM) induced contractions in isolated rat uterus and its modification by inhibitors of cAMP-dependent protein kinase (PKA) (Rp-cAMPS and TPCK), transcription (actinomycin D), protein synthesis (cycloheximide) and ornithine decarboxylase (alpha-difluoromethyl-ornithine, DFMO), and a polyamine (spermine) have been assayed. Forskolin (0.1 to 6 microM) induced concentration-dependent relaxation on KCl-induced tonic contractions in rat uterus (IC50: 0.55 +/- 0.12 microM) which was antagonized (p<0.05) by Rp-cAMPS (30 microM), TPCK (3 microM), cycloheximide (300 microM), actinomycin D (4 and 12 microM) and TPCK (3 microM) plus actinomycin D (12 microM). The IC50 values of forskolin in the presence of these drugs were 3.75 +/- 1.53 microM, 12.08 +/- 8.18 microM, 6.88 +/- 5.02 microM, 3.80 +/- 2.35 and 5.31 +/- 2.80 microM, and 4.26 +/- 3.65 microM respectively. Furthermore, DFMO (10 mM) also shifted the relaxation curve to forskolin to the right (IC50: 3.06 +/- 2.66 microM, p<0.05) but DFMO (10 mM) plus actinomycin D (12 microM) (IC50: 1.78 +/- 1.33 microM) did not. However, DFMO (10 mM) and actinomycin D (12 microM) did not antagonize the spermine (1-30 mM) elicited relaxation (IC50s: 7.8 +/- 0.7 mM vs 7.28 +/- 1.4 mM and 4.67 +/- 0.44 mM in the presence of DFMO and actinomycin D, respectively). Moreover, spermine (1 mM) did not decrease the forskolin induced relaxation and counteracted the antagonism produced by actinomycin D and DFMO. Our results suggest that, in rat uterus, forskolin: a) produced cAMP-dependent relaxation, as this is antagonized by Rp-cAMP and TPCK, and b) increased the activity of ornithine decarboxylase, as this is inhibited by DFMO. Therefore, polyamines could be the mediator of the cAMP-dependent transcriptional component involved in forskolin relaxation, since, as mentioned, DFMO antagonized this relaxation and spermine counteracted the displacement produced by DFMO and actinomycin D. Thus, a plasma membrane-nucleus interaction might, at least partially, explain the mechanisms involved in forskolin induced relaxation in smooth muscle of rat uterus under the present experimental conditions. PMID- 9416765 TI - Lamotrigine inhibits the in situ activity of tyrosine hydroxylase in striatum of audiogenic seizure-prone and audiogenic seizure-resistant Balb/c mice. AB - Lamotrigine (LTG), an anticonvulsive drug, was tested for its effects on striatal content of DA and its metabolites, DOPAC and HVA, in audiogenic seizure-resistant (ER) and audiogenic seizure-prone (EP) lines of Balb/c mice. A single dose of LTG (20 mg/kg) prevented audiogenic seizures in seizure-prone mice, while reducing substantially the striatal content of the DA metabolite, DOPAC (to less than 50% of saline-injected controls) in both seizure-resistant and seizure-prone mice. LTG administration also resulted in significant reduction of striatal content of HVA. The in situ activity of tyrosine hydroxylase (TH) in extracts of striatum was significantly reduced by LTG administration in both ER and EP mice. These data show that DA synthesis in the striatum of mice is substantially reduced by LTG administration. PMID- 9416764 TI - Enhancement of the small intestinal uptake of phenylalanylglycine via a H+/oligopeptide transport system by chemical modification with fatty acids. AB - The transport characteristics of chemically modified phenylalanylglycine (Phe Gly) with butyric acid (C4-Phe-Gly) and caproic acid (C6-Phe-Gly) were examined using rabbit intestinal brush-border membrane vesicles (BBMVs). In the presence of an inwardly H+ gradient (pH 7.5 inside, pH 6.0 outside), the uptake of Phe-Gly via BBMVs was significantly enhanced by the covalent attachment of butyric or caproic acid to the N-terminal of Phe-Gly. Moreover, C4-Phe-Gly uptake was stimulated by the trans-stimulation effect of some dipeptides and cefadroxil, and was inhibited by other dipeptides and cefadroxil. These results indicate that N terminal modified Phe-Gly with fatty acids are transported into BBMVs via an oligopeptide transporter. Therefore, chemical modification of dipeptides with fatty acids can enhance the intestinal absorption of dipeptide by a carrier mediated transport via an oligopeptide transporter. PMID- 9416766 TI - The effect of clenbuterol and recombinant erythropoietin on tumor growth and the anemia caused by the Walker 256 carcinosarcoma. AB - In patients with advanced cancer, anemia is a common complication indicative of a poor prognosis. Attempts to alleviate this have met with mixed success and interventions including erythropoietin often fail to elicit an appropriate response. We have used rats implanted with the Walker 256 carcinosarcoma as a model of non-responsive anemia. This study demonstrates that the provision of recombinant erythropoietin in the presence of clenbuterol, a beta2 agonist, attenuates both the cancer induced anemia and the growth of the tumor in this model. We hypothesize that this treatment relieves the tumor induced inhibition of hematopoiesis, which allows for not only an increase in hematocrit but an increased immunosurveillance resulting in tumor suppression. PMID- 9416767 TI - Diazepam alters caffeine-induced effects on beta-endorphin levels in specific rat brain regions. AB - The present study was conducted to evaluate the effects of caffeine and the benzodiazepine agonist diazepam, and a combination of both on beta-endorphin (beta-EN) levels in specific rat brain regions. Male Sprague-Dawley rats (150-200 g) adapted to a 12-hour light: 12-hour dark illumination cycle were used in this study. Caffeine (10 mg/kg), diazepam (2 mg/kg) or a combination of caffeine (10 mg/kg) and diazepam (2 mg/kg) were administered intraperitoneally to rats at 11:00 hr. Control animals were injected with saline. Animals were sacrificed by decapitation 1 h after injection, the brains were immediately removed; the cortex, hippocampus, hypothalamus and midbrain were dissected and their B-EN levels measured by radioimmunoassay. Caffeine administration significantly increased B-EN levels in the cortex. Similarly, administration of diazepam alone resulted in a significant increase of B-EN levels in cortex. However, concurrent administration of diazepam and caffeine resulted in higher increase of B-EN levels in cortex. No significant changes in B-EN levels were detected in hippocampus and midbrain after administration of either caffeine or diazepam alone. On the other hand, when diazepam and caffeine were concurrently administered a significant increase of B-EN levels were observed in the midbrain. Moreover, administration of diazepam alone resulted in a significant increase of B-EN levels in hypothalamus. This increase was still observed following concurrent administration of diazepam and caffeine. These results clearly indicate that diazepam alters caffeine-induced effects on B-EN in specific rat brain regions. PMID- 9416768 TI - The pituitary folliculo-stellate cell line TtT/GF augments basal and TRH-induced prolactin secretion by GH3 cell. AB - Numerous functions have been ascribed to anterior pituitary folliculo-stellate (FS) cells. Recently, we established a pituitary FS-like cell line (TtT/GF) from an isologously transplantable pituitary thyrotropic tumor. Here we investigated the effect of FS cells on prolactin (PRL) secretion by using a co-culture system between TtT/GF cells and GH3 cells. We observed that co-culture with TtT/GF cells significantly increased basal PRL secretion without affecting the growth rate of GH3 cells. Further, addition of anti-cytokine-induced neutrophil chemoattractant (CINC) antibody partially reduced PRL secretion in co-culture of GH3 cells with 10% TtT/GF cells. Co-culture with 5% TtT/GF cells also markedly enhanced the PRL response to thyrotropin-releasing hormone (TRH) by GH3 cells, while addition of anti-CINC antibody had no effects on the enhanced PRL response to TRH. These results suggest that FS cells augment basal PRL secretion and enhance the secretory response of PRL to TRH by a paracrine mechanism. PMID- 9416769 TI - Regulation of glutamate uptake in astrocytes continuously exposed to ethanol. AB - CNS glutamatergic transmission is altered by chronic ethanol intake and may underlie the behavioral hyperactivity associated with ethanol withdrawal. Because astrocytes regulate extracellular glutamate levels, the aim of this investigation was to characterize the effects of in vitro ethanol exposure on Na+-dependent glutamate uptake parameters in astrocytes. Ethanol exposure elicited a time and concentration-dependent increase in the maximal uptake capacity, Vmax, for [3H] glutamate, which was reversed upon withdrawal of ethanol from the media. None of the ethanol exposures had any effect on the Km for this process. In addition, the ethanol-induced increase in Vmax for glutamate was reversed by the protein kinase C inhibitors, calphostin C and bisindolylmaleimide, and was not associated with an increase in the expression of either of the major glutamate transporter proteins, GLT-1 or GLAST. PMID- 9416770 TI - Inhibition of shear stress-induced platelet aggregation by cilostazol, a specific inhibitor of cGMP-inhibited phosphodiesterase, in vitro and ex vivo. AB - Cilostazol(6-[4-(1-cyclohexyl-1H-tetrazol-5-yl)-butoxy]-3,4- dihydro-2(1H) quinolinone) selectively inhibits cGMP-inhibited phosphodiesterase (PDE3) and is a potent inhibitor of platelet aggregation induced by various agonists. Effect of cilostazol on shear stress-induced human platelet aggregation (SIPA) was examined in vitro and ex vivo. Cilostazol inhibited SIPA dose-dependently in vitro. The IC50 value of cilostazol for inhibition of SIPA was 15 +/- 2.6 microM (m +/- SE, n=5), which was very similar to that (12.5 +/- 2.1 microM) for inhibition of ADP induced platelet aggregation. Cilostazol potentiates the inhibition of SIPA by PGE1 and enhances its ability to increase cAMP concentrations. A single oral adminstration of 100 mg cilostazol to healthy volunteers produced a significant inhibition of SIPA. This study demonstrates that cilostazol is an effective inhibitor of SIPA, which may be important for the prevention and the treatment of arterial occlusive diseases. PMID- 9416771 TI - Chloroethylclonidine is a partial alpha1A-adrenoceptor agonist in cells expressing recombinant alpha1-adrenoceptor subtypes. AB - Chloroethylclonidine increased cytosol [Ca2+] in rat-1 fibroblasts stably expressing alpha1a-adrenoceptors. The effect of the imidazoline was dose dependent with a maximal effect (approximately 3-fold increase in [Ca2+]i) at 10 microM and it was blocked by phentolamine and 5-methyl urapidil, indicating that it was mediated through alpha1-adrenoceptors. Noradrenaline (1 microM) induced a much bigger effect (approximately 6-8-fold) in the same cells. When chloroethylclonidine was added before noradrenaline a dose-dependent inhibition of the effect of the natural catecholamine was observed. Chloroethylclonidine did not modified cytosol [Ca2+] in rat-1 fibroblast expressing alpha1b- or alpha1d adrenoceptors. However, the imidazoline acutely inhibited the effect of noradrenaline in these cells. It is concluded that chloroethylclonidine interacts with alpha1a-adrenoceptors as a partial agonist inducing Ca2+ mobilization in a very short time frame and that it is able to inhibit the action of noradrenaline when co-incubated with the catecholamine in cells expressing any of the three alpha1-adrenoceptor subtypes. PMID- 9416772 TI - Selective and non-selective ET antagonists reveal an ET(A)/ET(B) receptor mediated ET-1-induced antinociceptive effect in PAG area of mice. AB - The injection of endothelin-1 (ET-1) (2 pmol) into the dorsolateral periaqueductal gray area (PAG) of mice produces antinociceptive effect as underscored by increases in the latency time for the reaction to a hot plate. Pretreatment of the PAG area with bosentan (10 nmol) (a mixed ET(A)/ET(B) receptor antagonist), FR 139317 (5 nmol) (ET[A] receptor selective antagonist) or BQ-788 (5 nmol) (ET[B] receptor selective antagonist) greatly reduced the antinociceptive effect induced by ET-1. Therefore, ET-1 induces antinociceptive effects via both ET(A)/ET(B) receptors. In addition, since ET-antagonists lowered per se the control reaction time of the mice when administered alone to the PAG area, we would suggest that endogenous ET-1 acting within the PAG area contributes to the suppression of pain. PMID- 9416773 TI - Use of homology modeling in conjunction with site-directed mutagenesis for analysis of structure-function relationships of mammalian cytochromes P450. AB - In recent years, homology modeling has become an important tool to study cytochrome P450 function, especially in conjunction with experimental approaches such as site-directed mutagenesis. Molecular models of mammalian P450s can be constructed based on crystal structures of four bacterial enzymes, P450cam, P450 BM-3, P450terp and P450eryF, using molecular replacement or consensus methods. In a model built by molecular replacement, the coordinates are copied from those of a given template protein, while consensus methods utilize more then one protein as a template and are based on distance geometry calculations. The models can be used to identify or confirm key residues, evaluate enzyme-substrate interactions and explain changes in protein stability and/or regio- and stereospecificity of substrate oxidation upon residue substitution by site-directed mutagenesis. P450 models have also been utilized to analyze binding of inhibitors or activators, as well as alterations in inhibition and activation due to residue replacement. PMID- 9416774 TI - Brain histamine mediates the bombesin-induced central activation of sympatho adrenomedullary outflow. AB - Intracerebroventricular (i.c.v.) administration of bombesin (0.3 nmol) increased plasma levels of both adrenaline and noradrenaline in urethane anesthetized rats. These bombesin-induced increases were inhibited by i.c.v. pretreatment with pyrilamine, an H1-receptor antagonist. Ranitidine, an H2-receptor antagonist also inhibited the increase of adrenaline, however, its effective dose was much larger than that of pyrilamine. Furthermore, the bombesin-induced increase of noradrenaline was not effectively inhibited by ranitidine. In the next series, turnover of histamine was assessed by measuring accumulation of tele methylhistamine (t-MH), a major metabolite of brain histamine. I.c.v. administration of bombesin (0.3-3 nmol) increased turnover of hypothalamic histamine, while its intravenous administration was without effect. The present results suggest that the bombesin-induced central activation of sympatho adrenomedullary outflow is probably, at least in part, mediated through brain histaminergic neurons. PMID- 9416775 TI - Effects of L-dopa and bromocriptine on haloperidol-induced motor deficits in mice. AB - L-3,4-Dihydroxyphenylalanine (L-DOPA), the precursor of dopamine, and bromocriptine, a dopamine D2 receptor agonist, were investigated in haloperidol induced motor impairments in mice using both catalepsy and pole tests. In catalepsy test, subcutaneous treatment with haloperidol (0.125, 0.25 and 0.5 mg/kg) caused a cataleptic effect in mice in a dose-dependent manner. This cataleptic effect was evident upto 7 hr after haloperidol treatment. In pole test, haloperidol (0.125, 0.25 and 0.5 mg/kg) produced the prolongation of Tturn and TLA as a marker of bradykinesia in mice and the prolongation lasted at least 7 hr after haloperidol treatment. Intraperitoneal co-pretreatment with L-DOPA (400 mg/kg) + carbidopa (10 mg/kg) in mice decreased the catalepsy induced by haloperidol at a dose of 0.125 mg/kg, while co-pretreatment with L-DOPA (200 and 400 mg/kg) + carbidopa (10 mg/kg) dose-dependently decreased the haloperidol (0.125 mg/kg)-induced bradykinesia. The effect of LDOPA + carbidopa in pole test was more pronounced than that in catalepsy test. Intraperitoneal pretreatment with bromocriptine (2 and 4 mg/kg) in mice reduced the catalepsy and bradykinesia produced by haloperidol at a dose of 0.125 mg/kg. The effect of bromocriptine in pole test was relatively similar to that in catalepsy test. Also, co-pretreatment with LDOPA (400 mg/kg) + carbidopa (10 mg/kg) and pretreatment with bromocriptine (2 and 4 mg/kg) significantly decreased the catalepsy induced by haloperidol at a higher dose of 0.5 mg/kg. These results indicate that co-administration with L DOPA + carbidopa and single treatment with bromocriptine can decrease haloperidol induced catalepsy and bradykinesia in mice. Furthermore, our study suggests that pole test as well as catalepsy test is of value in the screening of drugs against neuroleptic-induced motor deficits. PMID- 9416776 TI - Free cortisol levels after awakening: a reliable biological marker for the assessment of adrenocortical activity. AB - In three independent studies, free cortisol levels after morning awakening were repeatedly measured in children, adults and elderly subjects (total n=152). Cortisol was assessed by sampling saliva at 10 or 15 minute intervals for 30-60 minutes, beginning at the time of awakening for two days (Study 1 and 2) or one (Study 3) day, respectively. In all three studies, free cortisol levels increased by 50-75% within the first 30 minutes after awakening in both sexes on all days. Premenopausal women consistently showed a stronger increase with a delayed peak after awakening compared to men on all days. In Study 2, there was a tendency for lower early morning free cortisol levels for women taking oral contraceptives (p=.10). Stability of the area under the curve (AUC) of the early morning free cortisol levels over the three (Study 1 and 2) or two (Study 3) days ranged between r=.39 and r=.67 (p<.001). Neither age, weight, nor smoking showed an effect on baseline or peak cortisol levels. Sleep duration, time of awakening and alcohol consumption also appeared to be unrelated to early morning free cortisol levels. From these data we conclude that in contrast to single assessments at fixed times, early morning cortisol levels can be a reliable biological marker for the individual's adrenocortical activity when measured repeatedly with strict reference to the time of awakening. PMID- 9416777 TI - Cytosolic estrogen receptor concentrations in the lumbosacral spinal cord fluctuate during the estrous cycle. AB - Estrogen responsive neurons have been anatomically identified with autoradiographic and immunohistochemical techniques and their distribution mapped in the lumbosacral spinal cord of female rats. Such neurons contain estrogen receptors (ERs). The present study was undertaken to: 1) quantify cytosolic estrogen receptor (ER) concentrations in the lumbosacral spinal cord and 2) determine if there is a relationship between cytosolic ER concentrations and fluctuations in serum estradiol (SE2) levels during the estrous cycle. Lumbosacral spinal segments were removed from intact cycling rats during the morning of proestrus, the afternoon of proestrus, and the morning of estrus, metestrus and diestrus. Trunk blood was collected at euthanasia and SE2 levels were determined using radioimmunoassay. Cytosolic ER concentrations were measured using a dextran-charcoal coated tube method. Concentrations of cytosolic ERs were low during estrus and metestrus, increased during diestrus with maximum concentrations during the afternoon of proestrus. These changes in ER concentrations paralleled SE2 levels measured in intact cycling animals; i.e., during estrus SE2 levels were low, but began to rise during metestrus, diestrus, and during the morning of proestrus with a maximum peak increase during the afternoon of proestrus. These data indicate there are fluctuations of cytosolic ER concentrations during the estrous cycle and that these changes coincide with changing SE2 concentrations suggesting that ER content is influenced by SE2. PMID- 9416778 TI - Genotype analysis of prepro-vasopressin signal peptide in vasopressin-producing and -non-producing lung tumors. AB - A polymorphism in the nucleic acid sequence encoding the signal peptide of the human prepro-vasopressin (AVP) has been reported in an AVP producing small cell lung carcinoma (SCLC) cell line. The difference predicts expression in tumor cells of a variant signal peptide with Pro for Leu 11. To clarify whether this difference is required for AVP secretion from SCLC cells and/or reflects increased mutagenesis in malignant tumors, the exon encoding the signal peptide of prepro-AVP in two AVP producing SCLC and 9 non-producing lung tumors was amplified using polymerase chain reaction. The variant sequence was neither found by direct sequencing nor by restriction enzyme analysis. These results suggest that similar to the hypothalamus the normal signal peptide is functional in tumor cells and that the variant signal peptide is not a prerequisite for AVP secretion from SCLC cells. PMID- 9416779 TI - A comparison of hepatic cytochrome P450 protein expression between infancy and postinfancy. AB - We immunochemically measured the contents of 9 different cytochrome P450 (CYP) isoenzymes expressed in the liver and compared them between two groups: one group of 6 infant and 4 perinatal patients and one group of 10 patients after infancy (over 1 year old). CYP protein expressed in human liver can be divided into three groups on the basis of expression pattern: (a) CYP2A6, 2C9, 2D6, 2E1, and 3A were present in all samples and no difference was observed between the two groups; (b) CYP1A2, 2B6, and 2C8 were expressed more after infancy than during infancy; and (c) CYP3A7, which has been considered a major CYP enzyme in fetal liver microsomes, was expressed in all infants as well as the four perinatal patients, whereas it was detected in only 2 patients after infancy. These results implied that CYP2A6, 2C9, 2D6, 2E1, and 3A are already expressed during perinatal and infant period, while CYP1A2, 2B6, and 2C8 are expressed highly in subjects over 1 year old, and CYP3A7 disappeared after infancy. PMID- 9416780 TI - Deficits in D-fenfluramine-sensitive pool of brain 5-HT following withdrawal from chronic cocaine exposure. AB - Recent head-twitch response (HTR) studies in mice have indicated that withdrawal from chronic cocaine exposure produces deficits in CNS conversion of L-tryptophan to 5-HT. In the present study, the ability of 5-HT releaser, d-fenfluramine, was utilized to induce the HTR in mice following abstinence from chronic cocaine exposure. d-Fenfluramine-induced HTR, is a 5-HT2A receptor-mediated phenomenon and its induction frequency can be regarded as an indirect but in vivo measure of basal brain 5-HT concentration. Thus, different groups of mice were injected with cocaine twice daily (0, 0.1, 0.5, 2.5, 5 or 10 mg/kg, i.p.) for either 7 or 13 days. At 24 h after last cocaine injection, the treated mice received d fenfluramine (5 mg/kg, i.p.) and the induced HTR (mean+/-SEM) was recorded for the next 30 min. Cocaine attenuated the d-fenfluramine-induced HTR frequency by 30-37% in the 13-day regimen and significant effects were observed from 0.5 mg/kg dose. At 24 h withdrawal in the 7-day cocaine exposure group, the mean HTR frequencies were attenuated, however, they did not achieve statistical significance. Extended abstinence studies (i.e. 24, 48, 72 and 96 h postwithdrawal) from chronic cocaine exposure (0, 0.5 and 5 mg/kg/day for either 7 or 13 days) indicated that in the 7-day exposure group, significant reductions (26, 39 and 22%) in HTR frequency occurred at 48, 72 and 96 h following withdrawal from 0.5 mg/kg cocaine, whereas its 5 mg/kg dose failed to induce a significant effect. In the 13-day exposure group significant reductions in HTR frequency were observed at 24 h abstinence (27%) for the 0.5 mg/kg cocaine dose and at 24 and 48 h for the 5 mg/kg. Overall, these results indicate that abstinence from chronic exposure to cocaine produces enduring deficits in basal 5 HT concentration. Lastly, serotonergic function appears to be uniquely sensitive to chronic administration of low doses of cocaine. PMID- 9416781 TI - [Tyr1]-nociceptin has naloxone-insensitive vasodilator activity in the hindquarters vascular bed of the rat. AB - The heptadecapeptide nociceptin, also known as Orphanin FQ, is a newly-discovered endogenous ligand for the opioid-like G-protein-coupled receptor ORL1. In the present study, in order to investigate the structure-activity relationship for nociceptin, responses to nociceptin, [Tyr1]-nociceptin, nociceptin-(2-17), nociceptin-(1-11), and nociceptin-(1-7) were compared in the hindquarters vascular bed of the rat. Injections of nociceptin (1-30 nmol), [Tyr1]-nociceptin (1-30 nmol), and met-enkephalin (10-300 nmol) induced dose-related decreases in hindquarters perfusion pressure, whereas injections of similar volumes of the saline vehicle had no effect. In terms of relative vasodilator activity, [Tyr1] nociceptin was similar to nociceptin, and these peptides were approximately 10 fold more potent than met-enkephalin in decreasing hindquarters perfusion pressure. In contrast, nociceptin-(2-17), nociceptin-(1-11), and nociceptin-(1-7) had no significant effect on hindquarters perfusion pressure when injected into the perfusion circuit in doses up to 100 nmol. The decreases in hindquarters perfusion pressure in response to [Tyr1]-nociceptin and nociceptin were not altered by the opioid receptor antagonist naloxone at a time when responses to met-enkephalin were reduced significantly. The results of the present study show that [Tyr1]-nociceptin and nociceptin have similar vasodilator activity in the hindquarters vascular bed and that responses to this novel nociceptin analog are not mediated by the activation of a naloxone-sensitive opioid receptor and are not dependent on the presence of the amino acid Phe at the N-terminus of the nociceptin sequence. Moreover, the results of the present study show that nociceptin-(2-17), nociceptin-(1-11), and nociceptin-(1-7) have no activity in the hindquarters vascular bed of the rat when injected in doses up to 100 nmol. PMID- 9416782 TI - The endogenous mu-opioid agonists, endomorphin 1 and 2, have vasodilator activity in the hindquarters vascular bed of the rat. AB - Endomorphin 1 and endomorphin 2 are newly-discovered endogenous ligands for the mu-opioid receptor. In the present study, responses to intra-arterial injections of endomorphin 1 and 2 were investigated in the hindquarters vascular bed of the rat. Under constant-flow conditions, endomorphin 1 and 2 induced dose-dependent decreases in hindquarters perfusion pressure when injected in doses of 3-100 nmol into the hindquarters perfusion circuit. Vasodilator responses to endomorphin 1 and 2 and met-enkephalin were attenuated by the opioid receptor antagonist naloxone (2 mg/kg i.v.) at a time when vasodilator responses to isoproterenol were not altered. In terms of relative vasodilator activity, endomorphin 1 and 2 were similar to ATP, 100-fold less potent than isoproterenol, and 10,000-fold less potent than acetylcholine. These data demonstrate that endomorphin 1 and 2 have significant naloxone-sensitive vasodilator activity in the hindquarters vascular bed of the rat. PMID- 9416783 TI - Efficient designs for studying synergistic drug combinations. AB - Distinguishing between pharmacologically additive and synergistic drug combinations requires experimental designs and statistical analyses that often require appreciable numbers of animals and much experimenter time. The current study employed a design in which individual dose-effect data from each drug were translated into theoretically additive total dose combinations, in a fixed drug proportion, in order to produce a composite additive dose-effect relation that could be compared with that of an actual mixture having the same proportion. Results from this approach, using a combination of intrathecal doses of morphine and clonidine, were virtually identical to those using isobolographic analysis of the same data set. Both analyses showed significant synergism for this combination and, in each method, it was not necessary to constrain the drug regression lines to parallelism. In contrast to the isobole approach, the use of the composite additive dose-effect relation also allows observation of the interaction over a range of effects while reducing the size of the data sets needed. PMID- 9416784 TI - Does dizocilpine (MK-801) inhibit the development of morphine-induced behavioural sensitization in rats? AB - Intermittent morphine pretreatment (10 mg/kg/day for 14 days) induced long lasting (one month post-treatment) sensitization to the locomotor effects of morphine and amphetamine in rats. Co-administration of the non-competitive NMDA receptor antagonist dizocilpine (MK-801) (0.1 mg/kg) with morphine did not prevent the development of long-term behavioural sensitization. However, this dose of MK-801 did cause long-term sensitization to its own locomotor effects. Co administration of 0.25 mg/kg MK-801 with morphine caused death in 60% of the animals. In the animals that survived MK-801 plus morphine pretreatment, neither short-term (3 days) nor long-term morphine-induced sensitization was observed. MK 801 alone (0.25 mg/kg/day for 14 days) induced short-term cross-sensitization to morphine. Thus, the development of long-term morphine-induced locomotor sensitization could only be prevented by a dose of MK-801 that yields a lethal combination with morphine. In addition, MK-801 induced sensitization to its own locomotor effects and cross-sensitization to morphine. These findings seriously question whether MK-801 can be used to study the development of morphine-induced behavioural sensitization. PMID- 9416785 TI - Three A's of repopulation during fractionated irradiation of squamous epithelia: Asymmetry loss, Acceleration of stem-cell divisions and Abortive divisions. AB - PURPOSE: To analyse the time-course and efficacy of repopulation in squamous epithelia, and to describe possible mechanisms of this regeneration response. MATERIAL AND METHODS: Experimental and clinical studies of repopulation in squamous epithelia have been reviewed to outline general features of repopulation during fractionated radiotherapy. RESULTS: Repopulation processes result in a relative increase in re-irradiation tolerance, which can be quantified as the dose equivalent compensated. CONCLUSIONS: If the stem-cell concept is accepted, changes in residual tissue tolerance must be based on net production of stem cells. For this, normal asymmetrical stem-cell divisions that render equal numbers of stem cells and differentiating daughters, must turn into symmetrical divisions with the production of two stem cells. Furthermore, the rate of change in residual tolerance indicates that the stem-cell proliferation rate is increased. In addition to these changes in the stem-cell proliferation pattern, a limited number of divisions by sterilized cells contributes to overall cell production and maintenance of tissue function. A valid model of repopulation in squamous epithelia hence must be based on three distinct mechanisms summarized as the three A's: Asymmetry loss, Acceleration of stem-cell divisions, and Abortive divisions. PMID- 9416786 TI - Effect of low-dose radiation on mouse dermal tissue using wound strength as an endpoint. AB - PURPOSE: To investigate the existence of enhanced sensitivity of dermal tissue to low radiation doses in the range of 0.1 Gy to 4 Gy/fraction. MATERIALS AND METHODS: Top-up technique of giving higher radiation doses to skin alone from 150 kVp X-rays followed by fractionated experimental doses to total body from 137Cs were given to C3H mice. Full-depth incisions were made and the tensile strength of skin from 14 day wounds were measured as a response of dermal tissue to radiation. RESULTS: There was no evidence of enhanced radiosensitivity to doses as low as 0.1 Gy. The data were better fitted by a nonparametric method that predicted the shape of survival curve better than an induced-repair model. CONCLUSIONS: Dermal tissue like spinal cord has a low sensitive fraction of cells, unlike jejunum, kidney or lung that showed sensitivity at these doses. Tissue kinetics play an important role in the use of low dose fractions to avoid injury to normal tissues. PMID- 9416787 TI - Induction of PBP74/mortalin/Grp75, a member of the hsp70 family, by low doses of ionizing radiation: a possible role in induced radioresistance. AB - The identification of genes whose expression is altered following exposure to a low dose of ionizing radiation (IR) is an important step in understanding the phenomenon of the adaptive response. Using the differential mRNA display method we have identified a gene whose expression is up-regulated following exposure to 0.25 Gy IR. Partial DNA sequence and restriction endonuclease analysis of this gene showed that it is identical to the gene encoding for the human peptide binding protein 74 (PBP74/mortalin/Grp75), a member of the heat shock 70 protein family. Time-course measurement of the PBP74/mortalin/Grp75 mRNA showed that its level was elevated after a lag of at least 15 min. The maximum induction appears to be at 30 min following gamma-irradiation and there is then a steady decline to control levels within 5 h in the HT29 cell line. On the other hand, the level of the PBP74/mortalin/Grp75 mRNA in the human breast adenocarcinoma cell line MCF-7 is consistently elevated after gamma-irradiation for up to 6 h post-irradiation. Furthermore, a cell line that does not demonstrate the induced radioresistance phenomenon (SW48) shows no induction of the PBP74/mortalin/Grp75 mRNA in contrast with HT29 or MCF-7. Treatment of the HT29 cells with antisense oligonucleotide directed towards the initiation codon of PBP74 sensitized cells to ionizing radiation. PMID- 9416788 TI - Chromosome aberration frequencies in human lymphocytes irradiated in a multi layer array by protons with different LET. AB - PURPOSE: To provide data on the dose-and linear energy transfer (LET)-dependence of the production of dicentrics in human lymphocytes. MATERIALS AND METHODS: Track segment irradiation with 16.5 MeV protons was performed in a multi-layer array allowing the simultaneous exposure of human peripheral lymphocytes in three successive samples. Within these samples the dose-averaged linear energy transfer (LD) of protons increased with increasing depth, i.e. LD = 3.5, 5.3 and 19.0 keV/microm. Dicentrics were scored in first division solid-stained metaphases. RESULTS: Dicentric yields measured in the first and second samples fit the linear quadratic model, those of the third sample fit a linear model of the dose response relationship. Relative to 137Cs gamma-rays, limiting RBE values of 2.9, 4.3 and 21.5 were determined from the ratios of the respective linear coefficients of the dose effect curves. The linear dose effect coefficient increases approximately proportionally with increasing LET. The quadratic coefficient is approximately constant for protons with LD of 3.5 and 5.3 keV/microm, and is not significantly different from zero at LD= 19.0 keV/microm. CONCLUSION: This result is well in line with theoretical predictions on the variations of dose-effect coefficients with radiation quality. However, such an evaluation can only be obtained from clearly defined track segment experiments. PMID- 9416789 TI - Relationship between PCC fragments and cell killing studied in X-irradiated CHO, CHO-K1 cells and two radiosensitive mutants xrs1 and xrs5. AB - PURPOSE: To investigate the correlation between PCC fragments and cell killing. MATERIALS AND METHODS: Induction and repair of DNA fragments were measured in CHO, CHO-K1, xrs1 and xrs5 cells using the premature chromosome condensation (PCC) technique and cell survival was determined by a colony assay. RESULTS: The number of PCC fragments measured in cells immediately fused after X-irradiation was the same for CHO (3.4 +/- 0.16/cell/Gy) and CHO-K1 (3.6 +/- 0.12/cell/Gy) cells but significantly higher for xrs1 (4.9 +/- 0.07/cell/Gy) and xrs5 cells (7.0 +/- 0.4/cell/Gy). The repair curve of PCC fragments studied for CHO, CHO-K1 and xrs5 cells was best described by a monophasic exponential decline with a final plateau; the half-time of this decline was always about 30 min. The number of unrejoined PCC fragments, which was measured 14h after irradiation, increased linearly with dose. The steepest increase was found for xrs5 cells (5.5 +/- 0.3 fragments per cell and per Gy), the lowest for CHO/CHO-K1 (0.9 +/- 0.1; 1.0 +/- 0.1) and for xrs1 in between (3.3 +/- 0.1). For all four cell lines the relationship between cell killing and unrejoined fragments could be described by a single curve with a D0 of 2.5 +/- 0.4 unrejoined PCC fragments per lethal event. CONCLUSIONS: The data showed that the number of unrejoined PCC fragments can be used as an indicator of cellular radiosensitivity. PMID- 9416790 TI - Oral platinum analogue JM216, a radiosensitizer in oxic murine cells. AB - This study was designed to compare radiosensitization by the oral platinum compound JM216 with cisplatin. RIF1 mouse tumour cells were treated at various doses and at various exposure times with JM216 and irradiated 15 min before the end of drug exposure. The fraction of cells surviving treatment was assessed by colony formation. Results were compared with those for equivalent treatments with cisplatin. JM216 alone showed exponential killing of RIF1 cells, being approximately three times less efficient than cisplatin on a molar basis. For radiosensitization studies, drug doses used gave approximately 50 or 90% cell killing alone. No radiosensitization was seen after 2-h drug exposures, but significant radiosensitization occurred after 1- and 0.5-h exposures (shorter times required proportionally higher drug doses, giving equivalent drug kill). The enhancement ratio and time dependence were similar for the two platinum compounds, reaching 1.5 at the highest concentrations tested. Drug DNA adduct formation was assessed using immunocytochemistry with the NKI-A59 antiserum raised to cisplatin-DNA adducts. The antiserum was shown to recognize JM216-DNA adducts in a dose-dependent manner and maximum nuclear staining was found to be correlated with cell kill for both drugs. However, neither the level of staining at the time of irradiation nor at the time of maximum adducts correlated with radiosensitization, indicating that the number of DNA adducts did not determine radiosensitization. Intracellular glutathione levels were shown to be decreased by the drug, but only by approximately 50%, implying that this was not the cause of the increased radiosensitivity. In summary, JM216 was shown capable of radiosensitizing a platinum-sensitive tumour line to an extent similar to cisplatin. Radiosensitization was exposure-time and drug-concentration dependent, but was not dependent on DNA adduct levels nor glutathione depletion. In contrast, cell kill after drug alone was well correlated with adduct levels. These data suggest that JM216 could replace cisplatin in combined radiotherapy chemotherapy studies, and also indicate that the NKI-A59 antibody could be used to monitor exposure levels in vivo. PMID- 9416791 TI - Effect of 6(5H)-phenanthridinone, a poly (ADP-ribose)polymerase inhibitor, and ionizing radiation on the growth of cultured lymphoma cells. AB - The ability of 6(5H)-phenanthridinone (Phen), a new potent poly(ADP ribose)polymerase (PARP) inhibitor, to potentiate the effect of ionizing radiation on tumour cells was evaluated. RDM4 murine lymphoma cells were irradiated using a 60Co panoramic source and then examined for their growth, cell cycle distribution and apoptosis. Phen (100 microM) was found to inhibit more than 90% of the PARP activity in control and irradiated cells. Cell proliferation was assessed using Alamar Blue, a new fluorometric assay. Phen was found to sharply increase the radiation-induced inhibition of cell proliferation. Indeed, at 2.5 Gy the relative cell number of Phen-treated cells was 60% below control levels. At the same radiation dose, the G2M arrest was also significantly reinforced by the addition of Phen. Furthermore, this PARP inhibitor was shown to significantly increase the amount of DNA fragmentation as revealed by the DNA migration pattern in agarose gel electrophoresis. Comparable results were obtained with 3-aminobenzamide, another PARP inhibitor, but at concentrations 200 fold higher. Taken together, these results indicate the potential interest of Phen as a valuable pharmacological probe for investigating the role of PARP in cellular responses to radiation. They also suggest a possible use of Phen as an adjuvant in radiotherapy. PMID- 9416792 TI - Intrinsic radiosensitivity of healthy donors and cancer patients as determined by the lymphocyte micronucleus assay. AB - The purpose of the study was to evaluate the usefulness of the cytokinesis-block micronucleus (MN) assay in assessment of radiosensitivity of lymphocytes in cancer patients. Lymphocytes from 15 cervical cancer patients, 21 head and neck cancer patients, seven lung cancer patients and 19 healthy donors were analysed using MN assay. The proportion of binucleate cells (BC) in cancer patients ranged from 22 to 56% and was significantly lower than in the control group (38-68%). MN frequency assessed five times over 6 months in four healthy donors showed that the interindividual variation was significantly higher than intraindividual. Before (0 Gy) and after irradiation (2 and 4 Gy) no statistical differences in the mean number of MN/BC were observed between healthy donors and cancer patient groups. Nevertheless, statistical cluster analysis allowed each group of donors to be divided into radioresistant and radiosensitive subgroups of patients. They showed significantly different dose response. Separate comparison of the mean MN frequency within all examined radioresistant and radiosensitive subgroups, showed statistically significant differences only after a dose of 4 Gy. At this dose, the lung cancer patients and cervical cancer patients from radiosensitive subgroups presented significantly higher radiosensitivity than the healthy donors. However, healthy donors from radioresistant subgroup did not differ significantly from cancer patients. This work has shown a high variation in interindividual radiosensitivity of donors and suggests the possibility of identifying radiosensitive patients on the basis of MN assay performed on lymphocytes. PMID- 9416794 TI - Estimates of neutron relative biological effectiveness derived from the Japanese atomic bomb survivors. AB - PURPOSE: To investigate neutron relative biological effectiveness. MATERIALS AND METHODS: The latest Japanese atomic bomb survivor cancer incidence and mortality datasets with the current (DS86) dosimetry system are analysed using generalized relative risk models and generalized absolute risk models, both with and without recently indicated adjustments to the Hiroshima DS86 neutron dose estimates. RESULTS: Without adjustments to the Hiroshima neutron doses, the best estimate of neutron relative biological effectiveness for all tumours in the incidence data is 63.3 (95% CI < 0-275.3) when a generalized relative risk model is used; when a generalized absolute risk model is used in the incidence data the best estimate is 53.5 (95% CI < 0-201.0); when a generalized relative risk model is used in the mortality data, the best estimate is 287.7 (95% CI 38.0- > 10[3]). When likely adjustments are made to the Hiroshima neutron doses the best estimate of neutron relative biological effectiveness in the incidence data using a generalized relative risk model is 15.1 (95% CI < 0-51.4); when a generalized absolute risk model is used in the incidence data the best estimate is 9.0 (95% CI < 0-32.9); when a generalized relative risk model is used in the mortality data the best estimate is 55.1 (95% CI 9.5-280.3). Although there are no significant differences between groupings of the solid tumour sites in their estimated neutron relative biological effectiveness, there are indications that the neutron relative biological effectiveness of solid tumours is lower than that of leukaemia, whether or not adjustments are made to the Hiroshima neutron dose estimates. Uncertainties in the likely adjustments to the DS86 Hiroshima neutron and gamma dose estimates as well as uncertainties in the modelling of excess risk in the two cities (Hiroshima and Nagasaki) imply that these findings should be treated with caution. CONCLUSIONS: Likely adjustments to the Hiroshima neutron dose estimates imply a substantial increase in information on neutron relative biological effectiveness. Whether or not adjustments are made to the Hiroshima neutron doses, there are indications of inconsistency between the estimates of neutron relative biological effectiveness for solid tumours and leukaemia. Dosimetric and modelling uncertainties mean that these findings should be treated with caution. PMID- 9416793 TI - Radiobiological evaluation of immigrants from the vicinity of Chernobyl. AB - Eighty individuals (55 adults and 25 children) who were residents of four cities (Kiev, Mozyr, Gomel and Bobrujsk) located 100-200 km from Chernobyl at the time of the accident in 1986 were tested after immigrating to the US from 1989-1991. A whole-body counter was employed to quantitate radiocesium content. In addition, two biological measures of radiation effects, namely, chromosomal integrity using the micronucleus assay and somatic mutation analysis of erythrocytes at the glycophorin A (GPA) locus, were applied to this group. Radiocesium activity in the body ranged from 0 to 56.8 Bq/kg with a mean and standard deviation of 5.0 +/ 8.2 and a median value of 2.0 Bq/kg. Mean radiocesium content by groups was highest in adult males (9.0 +/- 11.7; range 0.21-56.8 Bq/kg) followed by adult females (3.3 +/- 4.5; range 0-21.3 Bq/kg), male children (3.0 +/- 5.7; range 0 20.2 Bq/kg) and lowest in female children (1.6 +/- 3.5; range 0-12.7 Bq/kg). Individuals with the highest radiocesium content in each group belonged to one family that lived in Mozyr (100 km from Chernobyl) until emigrating in 1989. The frequency of lymphocyte micronuclei and erythrocyte GPA allele-loss (O/N) somatic mutations were both significantly correlated with radiocesium content (r=0.57, p=0.002; r=0.75, p=0.002, respectively). The micronucleus frequency also correlated with the estimated internal absorbed dose from radiocesium in a subset of 20 immigrants for whom this calculation was possible (r=0.71, p=0.0005). Altogether, the biomonitoring data indicate that some subjects had radiation doses sufficient to produce gene and chromosomal mutations in blood cells, although these effects cannot be attributed solely to radiocesium exposure. PMID- 9416795 TI - Chromosomal aberrations in blood lymphocytes of astronauts after long-term space flights. AB - PURPOSE: To analyse lymphocyte chromosomes from astronauts of MIR and EUROMIR missions. MATERIALS AND METHODS: Peripheral lymphocytes from seven astronauts before and after space flights were cultured and analysed for structural chromosomal aberrations. RESULTS: Chromosome-type but not chromatid-type aberrations were significantly elevated after space flights when compared to pre flight values. In one astronaut two rogue cells were found. CONCLUSIONS: Overall the frequencies of aberrations were found to be correlated with estimated absorbed cosmic radiation doses. PMID- 9416796 TI - Release of unaltered bases from polycrystalline pyrimidine DNA constituents after X-irradiation and bombardment with heavy ions. AB - PURPOSE: The radiation-induced release of unaltered bases from the lyophilized pyrimidine DNA constituents TMP, dCMP, CMP and dCyd was determined quantitatively in order to study explicitly the direct radiation effect. MATERIALS AND METHODS: X-irradiation under an air or a nitrogen gas atmosphere and heavy ion bombardment in the beam vacuum were employed at 300 K. The release of free bases was investigated using HPLC and 1H-NMR. Dose-yield curves for free radicals of X irradiated TMP and dCMP samples were determined by EPR spectroscopy. RESULTS: As one of the main products, the release of unaltered bases is linear with dose up to 360 kGy for X-irradiation or 200 kGy for heavy ion bombardment. The estimation of the radiation chemical yields revealed G values of about 10(-7) mol J-1. The heavy ion bombardment and X-irradiation under nitrogen effected a lower yield for base release than that for X-irradiation under air. For X-irradiation at sufficiently high doses about one order of magnitude difference in yields between bases released and free radicals formed was found. CONCLUSIONS: Compared with related findings as described in the literature, the G values from DNA are in the same order of magnitude, but from nucleosides in frozen aqueous solutions the yields of free bases are several orders of magnitude smaller. PMID- 9416797 TI - Hydrogen peroxide protects yeast cells from inactivation by ionizing radiation: a radiobiological paradox. AB - PURPOSE: To elucidate mechanisms of the interaction of hydrogen peroxide with chloride-derived cytotoxins under steady-state irradiation conditions and to determine the effects on cell viability. MATERIALS AND METHODS: Yeast cells were suspended in phosphate-buffered saline and exposed to 60Co gamma-irradiation under different conditions. The colony-forming ability was determined. RESULTS: Irradiation of PBS produces H2O2 and HOCl simultaneously. Under slightly acidic conditions and low oxygen tension the yield of HOCl exceeds that of H2O2 while at physiological pH and normoxic conditions H2O2 exceeds HOCl. Both substances react with each other rapidly in a pH-dependent way, even during an irradiation that lasts several seconds. As HOCl is about 1000-fold more toxic than H2O2 to the strain of Saccharomyces cerevisiae used in these experiments, it is evident that in an irradiation that produces more HOCl than H2O2 the radiation-induced damage will be large. If, in contrast, the cells are irradiated under conditions in which H2O2 production predominates, the damage will be small. One would therefore predict that addition of hydrogen peroxide to a cell suspension prior to irradiation should result in protection for suspended cells if H2O2 interferes with the generation of HOCl and thereby inactivates this more powerful toxin. Our data show that addition of H2O2 in sublethal concentration decreases radiation induced cell death to the level that is found in chloride-free solution, i.e. depending on pH, reduces it by a factor of > or = 3. PMID- 9416798 TI - Proliferation and cytogenetic studies in human blood lymphocytes exposed in vitro to 2450 MHz radiofrequency radiation. AB - Aliquots of human peripheral blood collected from two healthy human volunteers were exposed in vitro to continuous wave 2450 MHz radiofrequency radiation (RFR), either continuously for a period of 90 min or intermittently for a total exposure period of 90 min (30 min on and 30 min off, repeated three times). Blood aliquots which were sham-exposed or exposed in vitro to 150 cGy gamma radiation served as controls. The continuous wave 2450 MHz RFR was generated with a net forward power of 34.5 W and transmitted from a standard gain rectangular antenna horn in a vertically downward direction. The mean power density at the position of the cells was 5.0 mW/cm2. The mean specific absorption rate calculated by Finite Difference Time Domain analysis was 12.46 W/kg. Immediately after exposure, lymphocytes were cultured for 48 and 72 h to determine the incidence of chromosomal aberrations and micronuclei, respectively. Proliferation indices were also recorded. There were no significant differences between RFR-exposed and sham exposed lymphocytes with respect to; (a) mitotic indices; (b) incidence of cells showing chromosome damage; (c) exchange aberrations; (d) acentric fragments; (e) binucleate lymphocytes, and (f) micronuclei, for either the continuous or intermittent RFR exposures. In contrast, the response of positive control cells exposed to 150 cGy gamma radiation was significantly different from RFR-exposed and sham-exposed lymphocytes. Thus, there is no evidence for an effect on mitogen stimulated proliferation kinetics or for excess genotoxicity within 72 h in human blood lymphocytes exposed in vitro to 2450 MHz RFR. PMID- 9416799 TI - In vitro radiation-induced apoptosis and tumour response to radiotherapy: a prospective study in patients with non-Hodgkin lymphomas treated by low-dose irradiation. PMID- 9416800 TI - Neurosurgery for psychiatric disorders. PMID- 9416801 TI - Neurology and the bone marrow. PMID- 9416802 TI - Theodor Bilharz (1825-62). PMID- 9416803 TI - Pulmonary apex schwannoma. PMID- 9416804 TI - Hyperintensities of the anterior horn cells on MRI due to poliomyelitis. PMID- 9416805 TI - Contribution of mathematical modelling to the interpretation of bedside tests of cerebrovascular autoregulation. AB - OBJECTIVES: Cerebral haemodynamic responses to short and longlasting episodes of decreased cerebral perfusion pressure contain information about the state of autoregulation of cerebral blood flow. Mathematical simulation may help to elucidate which of the indices, that can be derived using transcranial Doppler ultrasonography and trends of intracranial pressure and blood pressure, are useful in clinical tests of autoregulatory reserve. METHODS: Time dependent interactions between pressure, flow, and volume of cerebral blood and CSF were modelled using a set of non-linear differential equations. The model simulates changes in arterial blood inflow and storage, arteriolar and capillary blood flow controlled by cerebral autoregulation, venous blood storage and venous outflow modulated by changes in ICP, and CSF storage and reabsorption. The model was used to simulate patterns of blood flow during either short or longlasting decreases in cerebral perfusion pressure. These simulations can be considered as clinically equivalent to a short compression of the common carotid artery, systemic hypotension, and intracranial hypertension. Simulations were performed in autoregulating and non-autoregulating systems and compared with recordings obtained in patients. RESULTS: After brief compression of the common carotid artery, a subsequent transient hyperaemia can be interpreted as evidence of intact autoregulation. During longlasting sustained hypoperfusion, a gradual increase in the systolic value of the blood flow velocity waveform along with a decrease in the diastolic value is specific for an autoregulating cerebrovascular system. CONCLUSION: Modelling studies help to interpret both clinical and experimental cerebral haemodynamic phenomena and their dependence on the state of autoregulation. PMID- 9416806 TI - Laboratory testing of the Spiegelberg brain pressure monitor: a technical report. AB - OBJECTIVES: The Spiegelberg brain pressure monitor is a low cost intracranial pressure monitoring system that has been used clinically for some time, mainly in Germany. To provide a rigorous bench comparison of the Spiegelberg monitor with the Camino pressure monitor an evaluation programme has been carried out in the UK Shunt Evaluation Laboratory. DESIGN: Drift over 72 hours and with temperature, a frequency response, and the accuracy of measurement of both static and pulsatile pressures have been tested simultanously in Camino and Spiegelberg transducers using a computerised rig. RESULTS: Long term zero drift was less than 0.7 mm Hg in both transducers. The Spiegelberg monitor showed no temperature drift whereas the Camino monitor had a drift of around 0.3 mm Hg/degrees C. The Spiegelberg monitor underread mean pressures <40 mm Hg by <1 mm Hg, but the error increased to 4.7 mm Hg at 100 mm Hg. The frequency bandwidth of the Spiegelberg monitor was 4 Hz at a low pressure. Underreading of the amplitude increased with the mean pressure, with a delay of about 0.1 s in the detection of the peaks of pulse waveform. CONCLUSION: The Spiegelberg transducer had excellent accuracy for static intracranial pressure measurement, but complex waveform analysis may be biased by its limited dynamic response. PMID- 9416807 TI - 1H magnetic resonance spectroscopy of chronic cerebral white matter lesions and normal appearing white matter in multiple sclerosis. AB - OBJECTIVES: To test the hypothesis that irrecoverable neurological deficit in multiple sclerosis is associated with axonal loss. METHODS: 1H magnetic resonance spectroscopy (MRS) was carried out in a group of patients with clinically definite multiple sclerosis (n=31). Using this technique, the apparent concentration of NA ([NA] the sum of N-acetyl aspartate (NAA), a neuronal marker, and N-acetylaspartylglutamate has been compared in four groups of patients with multiple sclerosis classified as relapsing-remitting, secondary progressive, primary progressive, benign, and a control group. RESULTS: In the patients with relapsing-remitting disease (n=9) there was a highly significant reduction of apparent NA (median 8.73 mM, range 6.86 mM-10.74 mM, P=0.0008) from an area of high signal compared with the control group (median 11.97 mM, range 10.55 mM-14.5 mM). In the patients with secondary progressive disease (n=10), there was again a highly significant reduction of apparent NA (median 7.82 mM, range 3.5 mM-10.3 mM, P=0.0003) from an area of high signal compared with the control group. In the patients with primary progressive disease (n=6) there was once again a highly significant reduction of apparent NA (median 8.83 mM, range 6.95 mM-9.89 mM, P<0.002) from an area of high signal compared with the control group. In the patients with benign disease, however, there was no significant difference in the apparent NA (median 10.5 mM, range 8.53 mM-12.8 mM, P>0.05) from an area of high signal compared with the control group. In the patients with benign disease (n=5) there was also no significant difference in the apparent NA (median 10.74 mM, range 8.58 mM-13.4 mM, P>0.3) from an area of normal appearing white matter compared with the control group. In the patients with primary progressive disease, however, there was a significant reduction of apparent NA from an area of normal appearing white matter (median 8.78 mM, range 8.7 mM-12.38 mM, P< 0.025) compared with the control group. There was a significant inverse correlation between [NA] from lesions in the patients with multiple sclerosis and disability as measured on the Kurtzke expanded disability scale score (r= -0.364, 0.05>P>0.02). CONCLUSION: These findings support the hypothesis that axonal loss is important in the development of disability in multiple sclerosis. They also provide evidence for axonal loss in normal appearing white matter in patients with primary progressive disease. PMID- 9416809 TI - Neuropathological assessment of the lesions of significance in vascular dementia. AB - OBJECTIVES: To better define the neuropathology of vascular dementia. METHODS: The neuropathological findings in 18 elderly, undemented subjects free of cerebrovascular disease were compared with 19 elderly undemented subjects who had cerebrovascular disease (many of whom had had a "stroke") and 24 elderly demented subjects who had cerebrovascular disease, but no other pathology to account for dementia. Cases in all groups were selected for absence or no more than very mild Alzheimer type pathology. RESULTS: Microvascular brain damage in the form of severe cribriform change and associated subcortical white matter damage and microinfarction were correlated with a history of dementia. Severe cribriform change was much more common and microinfarction somewhat more common in the demented group with vascular disease than the undemented group with vascular disease (P=0.0006 and P=0.031 respectively). Other findings of note were that congophilic angiopathy had a greater prevalence in the vascular dementia group than the control group, single cerebral infarcts were more common in the group who were undemented with vascular disease than in the group with dementia and vascular disease (P=0.0028), and the last group lacked evidence of macroscopic infarction more often than the first (P=0.034). There was a non-significant trend for the ratio of infarcted:uninfarcted tissue in one cerebral hemisphere to be higher in the group with dementia and vascular disease than in the group with vascular disease but no dementia. CONCLUSIONS: Microvascular disease, not macroscopic infarction, was the chief substrate of vascular dementia in this series of cases. PMID- 9416811 TI - Tetanus after cranial trauma in ancient Egypt. PMID- 9416810 TI - Decreased beta-phenylethylamine in CSF in Parkinson's disease. AB - OBJECTIVE: To determine the concentrations of beta-phenylethylamine (PEA) in CSF in patients with Parkinson's disease, and to evaluate the relation between concentration of PEA in CSF and severity of Parkinson's disease. METHODS: Using gas chromatography-chemical ionisation mass spectrometry, CSF concentrations of PEA were measured in 23 patients with Parkinson's disease (mean age, 64.0 (SD 8.2) years), of whom three were at Hoehn and Yahr stage II, 11 were at stage III, and nine were at stage IV. Comparison was made with eight patients with neuropathy (mean age, 57.0 (SD 19.2) years) and 12 controls without neurological disease (mean age, 57.6 (SD 4.8) years). RESULTS: Concentrations of PEA in CSF in Parkinson's disease were significantly lower (mean 205 (SD 131) pg/ml) than in patients with peripheral neuropathy (433 (SD 254) pg/ml) and controls (387 (SD 194) pg/ml). The concentrations of PEA in CSF correlated negatively with Hoehn and Yahr stage (P<0.01). CONCLUSIONS: There are decreased CSF concentrations of PEA in patients with Parkinson's disease. PMID- 9416808 TI - Comparison of [18F]FDG-PET, [99mTc]-HMPAO-SPECT, and [123I]-iomazenil-SPECT in localising the epileptogenic cortex. AB - OBJECTIVES: Firstly, to compare the findings of interictal 18F-fluorodeoxyglucose (FDG) positron emission tomography (PET) and of single photon emission computed tomography (SPECT) using 99mTc-hexamethyl propylene-amine-oxime (HMPAO) and 123I iomazenil in localising the epileptogenic cortex in patients who were candidates for epilepsy surgery, but in whom clinical findings, video EEG monitoring (V EEG), MRI, and neuropsychological evaluations did not give any definite localisation of the seizure onset. Secondly, to assess the ability of these functional methods to help in the decision about the epilepsy surgery. METHODS: Eighteen epileptic patients were studied with FDG-PET and iomazenil-SPECT. HMPAO SPECT was performed in 11 of these 18 patients. Two references for localisation was used--ictal subdural EEG recordings (S-EEG) and the operated region. RESULTS: Fifteen of 18 patients had localising findings in S-EEG. FDG-PET findings were in accordance with the references in 13 patients and iomazenil-SPECT in nine patients. HMPAO-SPECT visualised the focus less accurately than the two other methods. In three patients S-EEG showed independent bitemporal seizure onset. In these patients FDG-PET showed no lateralisation. However, iomazenil-SPECT showed temporal lobe lateralisation in two of them. CONCLUSION: FDG-PET seemed to localise the epileptogenic cortex more accurately than interictal iomazenil-SPECT in patients with complicated focal epilepsy. PMID- 9416812 TI - Event related potentials recorded in patients with locked-in syndrome. AB - OBJECTIVE: To determine the possibility of recording "cognitive" event related potentials (ERPs) in locked-in patients and therefore to determine whether ERPs can have a role in differential diagnosis of coma. METHODS: ERPs to classic auditory or visual "odd ball paradigms" were recorded three to four days, seven to eight days, and 30 to 60 days after admission to the intensive care unit, in four patients affected by basilar artery thromboembolism resulting in locked-in syndrome. Two patients (one 32 year old man, one 31 year old woman) could move the eyes laterally and vertically spontaneously and on command. One patient (a 39 year old man) had a "one and half syndrome", one patient (a 40 year old woman) could only elevate the left eyelid and eye. Results were compared with data from 30 age matched controls. In the last recording session a letter recognition paradigm was applied, in which ERPs were produced by the identification of letters forming a word. Results were compared with five age matched controls. Brainstem lesions extending to the pontomesencephalic junction were found on MRI and CT. RESULTS: ERPs to the oddball paradigms were recorded in three patients in the first recording session, in all patients in the second recording session. Latency, amplitude, and topographic distribution of ERP components were inside normal limits. With the letter recognition paradigm the patients could emit a P3 component to correspond with target letters, with the same margin of error as controls. CONCLUSION: It is possible to record ERPs in patients with locked-in syndrome shortly after the acute ischaemic lesion, and therefore to assess objectively cognitive activities. Furthermore the letter recognition paradigm could be implemented to facilitate linguistic communication with patients with locked-in syndrome. PMID- 9416813 TI - Long term effect of intravenous immunoglobulins and oral cyclophosphamide in multifocal motor neuropathy. AB - OBJECTIVES: To report the long term effect of the combined treatment with high dose intravenous immunoglobulins (i.v.Ig) and oral cyclophosphamide (CTX) in patients with multifocal motor neuropathy, and to determine whether the association of oral CTX in these patients may help to delay and, possibly, suspend i.v.Ig infusions. METHODS: Six patients with multifocal motor neuropathy responding to an initial course of i.v.Ig (0.4 g/kg/day for five consecutive days) were followed up for 37 to 61 (mean 47) months. All patients were subsequently treated with periodic i.v.Ig infusions (0.4 g/kg/day for two days at clinical worsening) and oral CTX (1-3 mg/kg/day). Improvement was assessed using the Rankin disability scale, a functional impairment scale for upper and lower limbs, and the MRC rating scale on the 20 most affected muscles. Electrophysiological and antiglycolipid antibody studies were performed before treatment, then yearly during follow up. RESULTS: All patients improved during treatment and, by the end of follow up or before worsening after therapy suspension, the median Rankin (P=0.0335) and upper (P=0.0015) and lower limb (P=0.0301) impairment scores and the mean MRC (P=0.0561) score were improved. By that time the number of nerves with partial motor conduction block was reduced (P=0.0197) and antiglycolipid antibody titres had decreased in all but one patient. All patients required periodic i.v.Ig infusions to maintain improvement but, after three to seven months of oral CTX, the interval between i.v.Ig infusions could be progressively prolonged until, in three patients, both treatments could be stopped for up to two years before clinical worsening. The main complications, both related to oral CTX, were haemorrhagic cystitis in two patients and persistent amenorrhea in one patient. CONCLUSIONS: I.v.Ig can induce and maintain improvement in multifocal motor neuropathy but does not eradicate the disease. Oral CTX may help to induce a sustained remission but it is not devoid of side effects and might therefore be reserved for patients with multifocal motor neuropathy who require frequent i.v.Ig infusions to maintain improvement. PMID- 9416814 TI - Neuromuscular disorder as a presenting feature of coeliac disease. AB - OBJECTIVES: To describe the range of neuromuscular disorders which may be associated with cryptic coeliac disease. METHODS: Nine patients were described with neuromuscular disorders associated with circulating antigliadin antibodies, whose duodenal biopsies later confirmed the diagnosis of coeliac disease. Neurological symptoms antedated the diagnosis of coeliac disease in all, and most had minimal or no gastrointestinal symptoms at the onset of the neuromuscular disorder. RESULTS: Three patients had sensorimotor axonal peripheral neuropathy, one had axonal motor peripheral neuropathy, one had probable inclusion body myositis and axonal motor peripheral neuropathy, one had polymyositis and sensorimotor peripheral neuropathy, one had mononeuropathy multiplex, one had neuromyotonia, and one had polyneuropathy. CONCLUSION: A wide range of neuromuscular disease may be the presenting feature of coeliac disease. This represents the first report of inclusion body myositis and neuromyotonia associated with coeliac disease. Estimation of circulating antigliadin antibodies should be considered in all patients with neuromuscular disease of otherwise obscure aetiology. PMID- 9416815 TI - The role of quantitative electromyography in inclusion body myositis. AB - OBJECTIVE AND METHODS: Inclusion body myositis is said to have both myopathic and neurogenic features on electrophysiological tests. Twenty one studies from 20 patients with biopsy defined inclusion body myosis, 13 of whom had quantitative electromyography (qEMG), were reviewed to determine if this technique added diagnostic specificity (one patient had both needle EMG and a later study with qEMG before muscle biopsy). RESULTS: Excessive numbers of polyphasic motor unit potentials (MUPs) (> 12% per muscle) were seen in 11 of the 13 patients. In 10 of 13 patients, mean MUP duration was abnormally reduced (26% to 48%). In three patients, mean MUP duration was abnormally reduced only after polyphasic MUPs were excluded. In all 13 patients, the simple MUP duration was reduced. Myopathy was unequivocally diagnosed in all 13 studies that included qEMG; of the remaining eight patients, the conclusions of the electrophysiological studies without qEMG was myopathy (one), neurogenic (four) or non-diagnostic (three). CONCLUSIONS: There is no evidence of a neurogenic component in inclusion body myosis if qEMG is used. Quantitative EMG is often necessary to make an electrophysiological diagnosis of a myogenic disorder in patients with inclusion body myosis. PMID- 9416816 TI - Trinucleotide (GAA)n repeat expansion in two families with Friedreich's ataxia with retained reflexes. AB - In occasional families in whom cases of classic Friedreich's ataxia (FRDA) coexist with affected cases with retained reflexes, linkage analysis has shown that both map to the FRDA locus on chromosome 9q13-21.1. A gene X25 has been identified within the critical region of the FRDA locus, and an intronic expanded GAA trinucleotide repeat has been found in most cases of FRDA. We report two further FRDA families in whom some patients with classic FRDA were areflexic whereas others had brisk reflexes. Molecular genetic analysis disclosed an abnormal trinucleotide repeat expansion within intron 1 of the FRDA gene in both phenotypes. PMID- 9416817 TI - Cerebral microembolism in patients with stroke or transient ischaemic attack as a risk factor for early recurrence. AB - The incidence of early recurrence in 32 patients who had had a transient ischaemic attack or stroke in the anterior circulation was studied. Patients with a potential cardiac source of embolism were excluded from the study. All patients had transcranial Doppler (TCD) monitoring of the symptomatic middle cerebral artery for microembolic signal detection within seven days from the onset of symptoms. Four patients had early recurrence during a mean follow up of 15 (SD11) days. All early recurrences occurred in the same arterial territory as the initial ischaemic event. Three of the four patients with early recurrence had prior microembolic signals. The incidence of early recurrence was 50% (3/6) in patients with microembolic signals and 3.8% (1/26) in patients without microembolic signals (P=0.02). The findings suggest that TCD monitoring of patients with recent cerebral ischaemia of presumed arterial origin allows recognition of a subset of patients at high risk for early recurrence. PMID- 9416818 TI - Clinical features and natural history of axial predominant adult onset primary dystonia. AB - The clinical features and natural history of 18 patients with adult onset axial predominant severe truncal primary dystonia are presented. The mean age of onset was 41 (42 for men, 39 for women) and there was a higher proportion of men (10:8). Analysis of their clinical features and follow up over three to five years or more showed that these patients generally conform to the characteristics of other types of adult onset primary dystonias. They tended to remain focal although there could be an initial contiguous spread, sometimes beginning in the craniocervical region and spreading axially or, rarely, vice versa. If spread occurred, involvement of the head, neck, and arms was mild in comparison with the severe dystonia of the trunk. However, in none of the patients with craniocervical or truncal onset did the dystonia spread to involve the legs. More than a third (seven of 18) of the patients had a prior history of injury at the site subsequently affected by dystonia. Treatment response to various drugs overall was poor but a third of the patients improved on treatment either with triple therapy (a combination of tetrabenazine, pimozide, and an anticholinergic drug) or high dose anticholinergic drugs alone. Severe depression occurred in 33% of patients, mainly due to the negative personal image arising from their disfiguring dystonia. None of the patients had a family history of dystonia and at the moment it is unclear whether these patients with sporadic axial dystonia are non-genetic phenocopies or are a manifestation of one or more of the genes that cause generalised dystonia or torticollis. PMID- 9416819 TI - Painful generalised clonic and tonic-clonic seizures with retained consciousness. AB - Two patients in whom consciousness and memory were retained during bilateral clonic or tonic-clonic seizures are reported on, and three patients reported on previously are reviewed. Ictal semiology differed from myoclonic and supplementary motor seizures, which are other seizure types characterised by bilateral motor movements and retained awareness. In the two new patients ictal pain was a prominent feature. It is proposed that propagation of seizure activity may be confined to the sensorimotor areas bilaterally while sparing the neural structures involved in maintaining consciousness and in processing language and memory. This unusual type of seizure may be misdiagnosed as a pseudoseizure. Detailed description of the ictal events and further laboratory evaluation including video-EEG monitoring may be necessary to make the distinction. PMID- 9416820 TI - Seasonal cyclothymia to seasonal bipolar affective disorder: a double switch after stroke. AB - The appearance of bipolar affective disorder after stroke depends on the presence of two factors: a predisposing factor of either genetic loading or subcortical atrophy, and a lesion of specific corticolimbic pathways involving the right hemisphere. Whether cyclothymia and seasonal affective disorder further predispose to poststroke affective disorder is not clear. A case is described which highlights these issues. The aetiological factors, pathophysiology, and diagnosis are discussed. PMID- 9416821 TI - Anosognosia: examining the disconnection hypothesis. AB - OBJECTIVE: To test the hypothesis that anosognosia for hemiparesis results from intrahemispheric disconnection. METHODS: Using right carotid barbiturate injection as a model for anosognosia for hemiparesis, systematic attempts were made to modify deficit awareness by providing the left hemisphere with explicit information regarding left upper extremity function. RESULTS: Experimental interventions failed to modify deficit awareness in 19 of 32 patients. In those patients who discovered their weakness, attempted movement of the weak limb seems more important than explicit observation of the extremity by the left hemisphere. CONCLUSIONS: The results fail to support Geschwind's disconnection hypothesis for anosognosia for hemiparesis. PMID- 9416822 TI - Dermatome variations in patients with transitional vertebrae. AB - OBJECTIVES: To test whether the position of lumbosacral dermatomes varies in the presence of transitional vertebrae. MATERIAL AND METHODS: Fifty consecutive male patients were tested for thoracolumbar and lumbosacral transitional vertebrae by radiography and for the position of the dermatome gap between the lumbar dermatomes L1, L2, L3, and the sacral dermatomes S2 and S3. The dermatome gap was documented with the use of the cremasteric reflex, the receptive field of which ends sharply at the gap which allows its delineation. RESULTS: Thirty two of 50 patients had a normal thoracolumbosacral spinal configuration, 10 patients had transitional vertebrae. The patients with a cranial displacement of the thoracolumbar or lumbosacral vertebral transition showed a dermatome gap which lay significantly more ventrally than in the patients with a normal spinal configuration. CONCLUSION: The finding supports the notion of a variant position of lumbosacral dermatomes in the presence of transitional vertebrae in males. PMID- 9416823 TI - Hereditary chin trembling or hereditary chin myoclonus? AB - Hereditary chin trembling is a rare autosomal dominant disease often considered as an "essential tremor variant". The clinical and neurophysiological data obtained in a new white family lead to the suggestion that this abnormal involuntary movement is a focal variant of hereditary essential myoclonus. PMID- 9416824 TI - Exacerbation of epilepsy by obstructive sleep apnoea. PMID- 9416825 TI - Fibrolipomatous hamartoma of the proximal ulnar nerve associated with macrodactyly and macrodystrophia lipomatosa as an unusual cause of cubital tunnel syndrome. PMID- 9416826 TI - Ipsilateral mydriasis in focal occipitotemporal seizures. PMID- 9416828 TI - Hereditary neuromyotonia: a mouse model associated with deficiency or increased gene dosage of the PMP22 gene. PMID- 9416827 TI - Parkinson's disease and tumour in the supplementary motor area: a re-evaluation. PMID- 9416829 TI - Anaphylactoid reaction to intravenous methylprednisolone in a patient with multiple sclerosis. PMID- 9416830 TI - Unilateral auditory hallucinations: ear or brain? PMID- 9416831 TI - Cu/Zn superoxide dismutase gene mutations in amyotrophic lateral sclerosis: correlation between genotype and clinical features. PMID- 9416833 TI - Colocalization of Ras and Ral on the membrane is required for Ras-dependent Ral activation through Ral GDP dissociation stimulator. AB - Ral GDP dissociation stimulator (RalGDS), a putative effector protein of Ras, stimulated the GDP/GTP exchange reaction of the post-tanslationally lipid modified but not the unmodified form of Ral in response to epidermal growth factor in COS cells. The RalGDS action on Ral was enhanced by an active form of Ras but not a Ras mutant which was not post-translationally modified in the cells. The RalGDS activity was inhibited by acidic membrane phospholipids such as phosphatidylinositol and phosphatidylserine but not by phosphatidylcholine or phosphatidylethanolamine in vitro. The post-translationally modified form but not unmodified form of Ras, Ral, and Rap were incorporated in liposomes consisting of these phospholipids. When Ral was incorporated alone in the liposomes, RalGDS did not stimulate the dissociation of GDP from Ral. When Ral was incorporated with the GTP-bound form of Ras in the liposomes, RalGDS stimulated the dissociation of GDP from Ral, while the GDP-bound form of Ras did not affect the RalGDS action. The Ras-dependent Ral activation through RalGDS required the Ras-binding domain of RalGDS. Rap, which shared the same effector loop as Ras, also stimulated the dissociation of GDP from Ral through RalGDS in the liposomes, although Rap did not enhance the RalGDS action in COS cells. Taken together with our previous observations that Ras recruits RalGDS to the membrane, these results indicate that the post-translational modifications of Ras and Ral are important for Ras dependent Ral activation through RalGDS and that colocalization of Ras and Ral on the membrane is necessary for Ral activation in intact cells. PMID- 9416832 TI - Inactivation of a c-Myb/estrogen receptor fusion protein in transformed primary cells leads to granulocyte/macrophage differentiation and down regulation of c kit but not c-myc or cdc2. AB - Primary murine fetal hemopoietic cells were transformed with a fusion protein consisting of the ligand-binding domain of the estrogen receptor and a carboxyl terminally truncated c-Myb protein (ERMYB). The ERMYB-transformed hemopoietic cells exhibit an immature myeloid phenotype when grown in the presence of beta estradiol. Upon removal of beta-estradiol, the ERMYB cells display increased adherence, decreased clonogenicity and differentiate to cells exhibiting granulocyte or macrophage morphology. The expression of the c-myc, c-kit, cdc2 and bcl-2 genes, which are putatively regulated by Myb, was investigated in ERMYB cells grown in the presence or absence of beta-estradiol. Neither c-myc nor cdc2 expression was down-regulated after removal of beta-estradiol demonstrating that differentiation is not a consequence of decreased transactivation of these genes by ERMYB. While bcl-2 expression was reduced by 50% in ERMYB cells grown in the absence of beta-estradiol, there was no increase in DNA laddering, suggesting that Myb was not protecting ERMYB cells from apoptosis. In contrast, a substantial (200-fold) decrease in c-kit mRNA level was observed following differentiation of ERMYB cells, and c-kit mRNA could be partially re-induced by the re-addition of beta-estradiol. Furthermore, a reporter construct containing the c-kit promoter was activated when cotransfected with a Myb expression vector, providing further evidence of a role for Myb in the regulation of c-kit. PMID- 9416835 TI - Protein kinase C-epsilon associates with the Raf-1 kinase and induces the production of growth factors that stimulate Raf-1 activity. AB - Several observations indicate that the Raf-1 kinase is a downstream effector of protein kinase C-epsilon (PKC epsilon). We recently have shown that Raf-1 is constitutively activated in PKC epsilon transformed Rat6 fibroblasts, and transformation can be reverted by expression of a dominant negative Raf-1, but not a dominant negative Ras mutant (Cacace et al., 1996). Cai et al. (1997) demonstrated that PKC epsilon induced proliferation of NIH3T3 cells is independent of Ras or Src, but depends on Raf-1. These authors further suggested that PKC epsilon activates Raf-1 by direct phosphorylation. Here we have investigated the functional interaction between PKC epsilon and Raf-1. PKC epsilon, but not PKC alpha, was found to bind to the Raf-1 kinase domain. The association appeared to be direct, as it could be reconstituted in vitro with purified proteins. Raf-1 and PKC epsilon could be co-precipitated from Sf-9 insect cells and PKC epsilon transformed NIH313 cells (NIH/epsilon). The association was negatively regulated by ATP in vitro and by TPA treatment in NIH/epsilon cells, but not in Sf-9 insect cells. Raf-1 was constitutively activated in NIH/epsilon cells. However, using coexpression experiments in Sf-9 cells and transiently transfected A293 cells we did not obtain any evidence for a direct activation of Raf-1 by PKC epsilon. PKC epsilon did not induce translocation of Raf-1 to the membrane. Furthermore, PKC epsilon did not activate Raf-1 nor enhance the kinase activity of Raf-1 that had been pre-activated by coexpression of Ras or the Lck tyrosine kinase. In contrast, conditioned media from PKC epsilon transformed cells induced a robust activation of Raf-1. This activation could be partially reproduced by recombinant TGFbeta, a growth factors secreted by PKC epsilon transformed Rat6 cells. In conclusion, our results suggest that PKC epsilon stimulates Raf-1 indirectly by inducing the production of autocrine growth factors. PMID- 9416834 TI - Tyrosine kinase activity of the EGF receptor is enhanced by the expression of oncogenic 70Z-Cbl. AB - The 120 kD product of the c-Cbl oncogene is a prominent substrate of protein tyrosine kinases that lacks a known catalytic activity but possesses an array of binding sites for cytoplasmic signalling proteins. An oncogenic form of Cbl was recently identified in the 70Z/3 pre-B cell lymphoma which has a small deletion at the N-terminus of the Ring finger domain. This form of Cbl, termed 70Z-Cbl, exhibits an enhanced level of tyrosine phosphorylation compared with c-Cbl. Here we demonstrate that the expression of 70Z-Cbl induces a tenfold enhancement in the kinase activity of the EGF receptor in serum-starved and EGF-stimulated cells. In serum-starved cells this results in EGF receptor autophosphorylation and the recruitment of Grb2, Shc and Sos1 but does not induce a corresponding increase in MAP kinase activity. Furthermore the expression of 70Z-Cbl greatly enhances EGF-induced tyrosine phosphorylation of the protein tyrosine phosphatase SHP-2. We also show that the Cbl/EGF receptor complex is predominantly associated with CrkII and is distinct to the Grb2/Shc/Sos1 complex that associates with the EGF receptor. These findings therefore demonstrate a biochemical effect of an oncogenic Cbl protein and support predictions from C. elegans that Cbl functions as regulator of receptor tyrosine kinases. PMID- 9416836 TI - An alternative pathway for expression of p56lck from type I promoter transcripts in colon carcinoma. AB - The lymphoid-specific protein tyrosine kinase, p56lck which is essential for both T cell development and function, is aberrantly expressed in colon and small lung carcinoma lines. In this paper, we demonstrate p56lck is also expressed in colon tumour biopsies due predominantly or exclusively to the use of the lck type I promoter. In T leukaemia lines, the lck type I promoter requires binding sites for both Ets- and Myb-related transcription factors. In contrast, in colon tumour lines the activation of the lck type I promoter requires the Ets but not the Myb binding site. In these lines, a consensus binding site for HMG-related transcription factors, AACAAAG, is required for efficient lck type I promoter activity. Sox-4 is a candidate transcription factor for binding and activating the lck type l promoter in colon carcinoma cells. Co-expression of Ets-1 and Sox 4, but neither protein alone, was sufficient to activate the lck type l promoter in HeLa cells which do not normally express lck transcripts. These results suggest that aberrant expression of p56lck from the lck type l promoter in colon carcinoma arises from transcriptional activation mediated by Ets- and HMG-related transcription factors. PMID- 9416837 TI - The Myb leucine zipper is essential for leukemogenicity of the v-Myb protein. AB - The AMV v-Myb oncoprotein causes oncogenic transformation of myelomonocytic cells in vivo and in vitro. Its transforming capacity is strictly dependent upon the N terminal DNA binding domain, the central transactivation region, and on the C terminal domain containing a putative leucine zipper motif. Here we show that the v-MybL3,4A mutant, in which Leu325 and Leu332 of the leucine zipper have been replaced by alanines, failed to induce leukemia in virus infected chicken. This demonstrates that the leucine zipper domain is indispensable for v-myb induced leukemogenesis in vivo. v-MybL3,4A was, however, still able to transform myelomonocytic cells from chicken bone marrow in vitro. Yet, while v-mybL3,4A transformed cells were impaired in growth at 37 degrees C, they failed to grow at 42 degrees C, the physiological body temperature of avian species. This might explain the loss of v-MybL3,4A leukemogenic potential in vivo. We also demonstrate that the v-Myb leucine zipper domain interacts in vitro with two host cell proteins, p26 and p28. This interaction is compromised in v-MybL3,4A indicating that binding of v-Myb to p26 and p28 might be important for the leukemogenic potential of v-Myb. PMID- 9416838 TI - TP53 mutational pattern in Spanish and Polish non-small cell lung cancer patients: null mutations are associated with poor prognosis. AB - Inactivation of TP53 tumor suppressor gene is the most frequent molecular alteration in NSCLC, involving up to 60% of cases. Furthermore, TP53 mutational spectrum is related to the type of mutagen exposure, as well as racial and/or diet differences. Nearly 95% of TP53 perturbations affect codons included within exons 5-8 which encode for almost the entire DNA-binding domain. In this study we addressed the possible prognostic value of the molecular alterations identified in exons 5-8 of the TP53 gene in DNAs from 151 paraffin-embedded NSCLC sections corresponding to 59 Spanish and 92 Polish stage I-IIIA resected patients. PCR/single-strand conformation polymorphism (SSCP) analysis revealed that the occurrence of TP53 exon 5-8 mutations was 17/59 (29%) in the Spanish cohort and 17/92 (18%) in the Polish group. However, when DNA sequencing analysis was performed, these frequencies were reduced because of the presence of SSCP-false positive, intronic and silent mutations and polymorphisms. Fifteen of the 59 Spanish NSCLC tumors (25%) harbored TP53 mutations affecting exons 5-8 coding sequences, whereas only 12 of 92 Polish neoplasms (13%) contained alterations in the central hydrophobic region of p53. Our results indicate that the occurrence of TP53 mutations affecting exon 5-8 coding sequences in some European NSCLC populations may be lower than previously reported, and that the TP53 mutational patterns of these cohorts differ somewhat. The Spanish NSCLC patients contained missense mutations (9/59, 15%) and a relatively high percentage of null mutations (5/59, 8%) while the Polish patients mostly harbored missense mutations (9/92, 10%) and only one tumor contained a null type (1/92, 1%). Moreover, most TP53 missense mutations in the Spanish group were located outside the conserved regions, whereas the same mutations in the Polish group affected conserved amino acids. Furthermore, the Polish patients harbored a high percentage of G-->A transitions (most of them at non-CpG sites), while G-->T transversions were predominant in the Spanish group. Our findings suggest that there may be different racial or exogenous factors in these two populations which may help to explain both the distinct TP53 mutational pattern and the lower frequency obtained in the Polish group. The presence of missense mutations did not confer a worse clinical outcome in these subsets of NSCLC patients. However, patients whose tumors contained null TP53 gene mutations had a 5 month median disease-free survival time in contrast with 42 months in those patients without mutations (P=0.008). These findings suggest that loss of p53 function may enhance tumor progression in NSCLC patients independently of whether dominant negative TP53 missense mutations are present. PMID- 9416839 TI - Membrane recruitment of Raf-1 is not the only function of Ras in Raf-1 activation. AB - Ras interacts with Raf-1 and stimulates its kinase activity, which results in activation of the mitogen-activated protein (MAP) kinase cascade. It has been proposed that the main function of Ras in Raf-1 activation is to recruit Raf-1 to the plasma membrane, where a separate activation event such as phosphorylation takes place. Here, we examined the activities of various mutants of human Ha-Ras to induce membrane translocation of Raf-1 and to activate Raf-1 in vivo. Overexpression of an activator region mutant Ha-Ras(V45E) in COS7 cells induced membrane translocation of Raf-1 as effectively as wild-type Ha-Ras. However, the activity of this mutant to activate Raf-1 and extracellular signal-regulated kinase-2 (ERK2) was attenuated by approximately 70% compared to that of wild-type Ha-Ras. The decrease in the specific activity was further demonstrated by measuring the activity of the Ha-Ras(V45E)-associated Raf-1 purified from the membrane fraction. These results imply that the association of Ras with Raf-1 has another important consequence, presumably dependent on the interaction between its activator region and Raf-1, than the simple recruitment of Raf-1 to the plasma membrane. PMID- 9416840 TI - Rel/NF-kappa B transcription factors and I kappa B inhibitors: evolution from a unique common ancestor. AB - From the sequences of Rel/NF-kappa B and I kappa B proteins, we constructed an alignment of their Rel Homology Domain (RHD) and ankyrin repeat domain. Using this alignment, we performed tree reconstruction with both distance matrix and parsimony analysis and estimated the branching robustness using bootstrap resampling methods. We defined four subfamilies of Rel/NF-kappa B transcription factors: (i) cRel, RelA, RelB, Dorsal and Dif; (ii) NF-kappa B1 and NF-kappa B2; (iii) Relish and (iv) NF-AT factors, the most divergent members. Subfamilies I and II are clustered together whereas Relish diverged earlier than other Rel/NF kappa B proteins. Three subfamilies of I kappa B inhibitors were also defined: (i) NF-kappa B1 and NF-kappa B2; (ii) close to subfamily I, the short I kappa B proteins I kappa B alpha, I kappa B beta and Bcl-3; (iii) Relish that diverged earlier than other I kappa B inhibitors. Our definition of groups and subfamilies fits to structural and functional features of the Rel/NF-kappa B and I kappa B proteins. We also showed that ankyrin repeats of NF-kappa B1, NF-kappa B2 and Relish are short I kappa B-specific ankyrin motifs. These proteins defining a link between Rel/NF-kappa B and I kappa B families, we propose that all these factors evolved from a common ancestral RHD-ankyrin structure within a unique superfamily, explaining the specificities of interaction between the different Rel/NF-kappa B dimers and the various I kappa B inhibitors. PMID- 9416841 TI - The CD2-scl transgene alters the phenotype and frequency of T-lymphomas in N-ras transgenic or p53 deficient mice. AB - Abnormal expression of SCL (TAL-1/TCL5) occurs in the majority of paediatric cases of acute T-cell lymphoblastic leukemia (T-ALL). Unexpectedly however, transgenic mice carrying scl coupled to the human T-cell specific CD2 enhancer (CD2-scl) did not spontaneously develop T-cell lymphomas despite high levels of scl expression in their thymocytes. Analogous to other transgenic models of lymphomagenesis, it is likely that additional genetic abnormalities are required to cooperate with scl to trigger lymphomagenesis. Two possible candidates are the p53 and N-ras genes which are mutated in some cases of T-ALL, particularly in relapsed disease. Therefore, we examined lymphomagenesis in the progeny of CD2 scl mice crossed with N-ras transgenic mice or p53 deficient. Surprisingly, the frequency of lymphomas in the p53 nullizygous or N-ras transgenic mice was not enhanced by expression of the scl transgene. In fact, expression of scl in both genetic backgrounds paradoxically reduced the frequency of thymic lymphomas and, at least in the p53 nullizygous mice, shifted the pattern of organ involvement to the peripheral lymphoid organs. In contrast, CD2-scl transgene expression accelerated lymphomagenesis in p53 heterozygous mice. These data suggest that the collaborative effects of scl with N-ras or p53 vary according to the developmental stage of the T-cell. PMID- 9416842 TI - Changes in p53 expression can modify cell shape of ras-transformed fibroblasts and epitheliocytes. AB - p53 plays an important role in restriction of abnormal cell proliferation. Loss of this safeguard function induced by p53 mutations seems to be a key mechanism in oncogenesis. It cannot be excluded however, that in addition to elimination of p53-dependent checkpoints and/or apoptosis p53 mutations may cause additional effects that contribute to oncogenic transformation. In order to analyse the effects of wild-type (wt) and mutant p53 on expression of ras-induced morphological transformation we used the method of computer-assisted morphometry. The following parameters were determined: a) the area covered by the spread cells; b) dispersion and c) elongation of cell contours. The last two indices characterise cell shape. Elongation indicates the degree of bipolarity of cell contour and dispersion-the degree of its multipolarity. Transformation of Rat1 and mouse 10(3) fibroblasts by N-rasasp12 oncogene was accompanied by dramatic decrease of cell area and increase of dispersion and elongation. IAR-2 discoid epitheliocytes expressing exogenous ras oncogene transformed into polarised cells with decreased cell area. Fluorescent microscopic examination of actin cytoskeleton stained with rhodamine-phalloidin had shown that ras-induced transformation of IAR-2 cells is characterised by disappearance of circumferential actin bundle and straight fibers. Neither did we reveal actin stress-fibers in the ras-transformed Rat1 cells. Transduction of p53 cDNAs caused no significant changes in morphometric parameters of non-transformed parental Rat1, IAR-2 and 10(3) cells, but some of the p53 mutants modified cell shape of ras-transformed cells. p53-His273, unlike other tested p53 mutants (Tyr141, His194, Trp248), induced partial reversion of morphological transformation in both Rat1 fibroblasts and IAR-2 epitheliocytes. Its expression led to increase of average cell area, decrease of dispersion and elongation indices, and re appearance of actin bundles. Exogenous wild-type p53 also caused some reversion of transformed phenotype of Rat/ras cells, but its effect was weaker than that of the p53-His273. In contrast, another p53 mutant p53-His175 was able to enhance ras-induced morphological transformation in p53-deficient murine 10(3) fibroblasts that is consistent with possible involvement of some gain of function activity of mutant p53 in modulation of cell shape. Possible pathways that might be responsible for p53-induced changes of cell morphology are discussed. PMID- 9416843 TI - p27Kip1 overexpression causes apoptotic death of mammalian cells. AB - p27KiP1, a member of the Cip/Kip family of cyclin-dependent kinase (cdk) inhibitors, has been implicated in mediating G1 arrest in response to a variety of growth inhibitory signals. Its importance in regulating cell growth is emphasized by the fact that mice lacking p27Kip1 are abnormally large and display hyperplasia of multiple tissues. However, these mice retain the ability to undergo G1 arrest in response to growth inhibitory signals, suggesting that p27KiP1 may serve other functions important for controlling tissue growth. In the present study, we utilized an adenoviral vector-based expression system to examine the consequences of p27Kip1 overexpression in the human carcinoma cell lines A549, HeLa and RKO, in human melanoma SK-MEL-110 cells, in human lung fibroblasts IMR90 and in the rat fibroblast line Rat1. We demonstrate that overexpression of p27Kip1 leads to apoptotic cell death in all cell types, and further show that ectopic expression of Bcl-2 can protect HeLa cells from apoptosis mediated by p27Kip1 overexpression. To our knowledge, this is the first study demonstrating that p27Kip1 can induce apoptosis. Our findings provide new insight into the possible functions of this growth regulatory protein, and support the potential utility of gene therapeutic approaches aimed at elevating p27Kip1 expression for treatment of human cancers. PMID- 9416844 TI - Analysis of the CDKN2A, CDKN2B and CDK4 genes in 48 Australian melanoma kindreds. AB - Germline mutations within the cyclin-dependent kinase inhibitor 2A (CDKN2A) gene and one of its targets, the cyclin dependent kinase 4 (CDK4) gene, have been identified in a proportion of melanoma kindreds. In the case of CDK4, only one specific mutation, resulting in the substitution of a cysteine for an arginine at codon 24 (R24C), has been found to be associated with melanoma. We have previously reported the identification of germline CDKN2A mutations in 7/18 Australian melanoma kindreds and the absence of the R24C CDK4 mutation in 21 families lacking evidence of a CDKN2A mutation. The current study represents an expansion of these efforts and includes a total of 48 melanoma families from Australia. All of these families have now been screened for mutations within CDKN2A and CDK4, as well as for mutations within the CDKN2A homolog and 9p21 neighbor, the CDKN2B gene, and the alternative exon 1 (E1beta) of CDKN2A. Families lacking CDKN2A mutations, but positive for a polymorphism(s) within this gene, were further evaluated to determine if their disease was associated with transcriptional silencing of one CDKN2A allele. Overall, CDKN2A mutations were detected in 3/30 (10%) of the new kindreds. Two of these mutations have been observed previously: a 24 bp duplication at the 5' end of the gene and a G to C transversion in exon 2 resulting in an M531 substitution. A novel G to A transition in exon 2, resulting in a D108N substitution was also detected. Combined with our previous findings, we have now detected germline CDKN2A mutations in 10/48 (21%) of our melanoma kindreds. In none of the 'CDKN2A negative' families was melanoma found to segregate with either an untranscribed CDKN2A allele, an R24C CDK4 mutation, a CDKN2B mutation, or an E1beta mutation. The last three observations suggest that these other cell cycle control genes (or alternative gene products) are either not involved at all, or to any great extent, in melanoma predisposition. PMID- 9416845 TI - Treatment of Wegener's granulomatosis: time to change the standard of care? PMID- 9416846 TI - Th1/Th2 cytokine balance in arthritis. PMID- 9416847 TI - Winners of the 1997 American College of Rheumatology slide competition and presentation of the osteoporosis supplement to the Clinical Slide Collection on the Rheumatic Diseases. ACR Audiovisual AIDS Subcommittee. PMID- 9416848 TI - Ribozyme-based gene cleavage approach to chronic arthritis associated with human T cell leukemia virus type I: induction of apoptosis in synoviocytes by ablation of HTLV-I tax protein. AB - OBJECTIVE: To develop gene therapy for patients with human T cell leukemia virus type I (HTLV-I)-associated arthropathy (HAAP), we investigated the effects of ribozyme-mediated cleavage of HTLV-I tax/rex messenger RNA (mRNA) on synovial overgrowth. METHODS: We introduced 2 hammerhead ribozymes targeted against HTLV-I tax/rex mRNA into synovial cells obtained from patients with HAAP and from patients with HTLV-I-negative rheumatoid arthritis (RA) and examined the ribozyme mediated ablation of Tax expression. Using standard methods, we also determined the cells' ability to stop proliferating and to undergo apoptosis. RESULTS: The ribozymes successfully cleaved tax/rex mRNA in HAAP patient synoviocytes. Both tax mRNA expression and Tax protein synthesis were inhibited significantly, resulting in inhibition of synovial cell growth and induction of apoptosis. In contrast, synovial cells from RA patients were not affected. CONCLUSION: In vitro results suggest that ribozyme-mediated gene therapy can inhibit the growth of HTLV-I-infected synovial cells, which is maintained by Tax protein, in HTLV-I related diseases including HAAP. PMID- 9416849 TI - Modulation of synovial cell function by somatostatin in patients with rheumatoid arthritis. AB - OBJECTIVE: To elucidate the role of neurologic, endocrine, and immune system interactions in the development of pathologic responses in patients with rheumatoid arthritis (RA), we studied somatostatin (SOM) production and somatostatin receptor (SOMR) expression in RA synovium and its function in patients with RA. METHODS: The effects of SOM on proinflammatory cytokine (interleukin-6 [IL-6] and IL-8) and collagenase production by RA synovial cells were estimated by enzyme-linked immunosorbent assay, and their messenger RNA expression was assessed by reverse transcription-polymerase chain reaction (RT PCR) using limiting dilutions of the complementary DNA. The expression of SOMR by RA synovial cells was also studied by RT-PCR. Local production of SOM was estimated by RT-PCR and immunohistochemical staining. RESULTS: Physiologic concentrations (approximately 10(-10)M) of SOM inhibited proliferation of RA synovial cells. The production of proinflammatory cytokines and matrix metalloproteinases by RA synovial cells was also modulated by SOM. SOMR subtypes 1 and 2 were expressed on fibroblast-like synovial cells, and the expression of SOMR-2 was up-regulated by proinflammatory cytokine treatment of the synovial cells from patients with RA. RA fibroblast-like cells synthesized SOM by themselves, suggesting that SOM acts as an autocrine regulator of synovial cell function in patients with RA. CONCLUSION: SOM inhibited aberrant synovial cell function in patients with RA, suggesting possible clinical applications of this neuropeptide. PMID- 9416850 TI - Oncostatin M stimulates monocyte chemoattractant protein-1- and interleukin-1 induced matrix metalloproteinase-1 production by human synovial fibroblasts in vitro. AB - OBJECTIVE: To measure levels of oncostatin M (OSM) in the synovial fluid of rheumatoid arthritis (RA) patients and to examine the activities of human OSM in the regulation of human synovial fibroblast (HSF) production of chemokines and matrix metalloproteinases (MMP-1 and MMP-3) in vitro. METHODS: We examined the levels of OSM in the synovial fluids of patients with arthritis by an enzyme linked immunosorbent assay (ELISA). ELISA of cell culture supernatants and Northern blots were used to assess responses of HSF to interleukin-1alpha (IL 1alpha), OSM, and other members of the IL-6/leukemia inhibitory factor (IL-6/LIF) family of cytokines. RESULTS: We detected variable levels of OSM antigen in 9 of 10 RA patient synovial fluids, but levels were not detectable in 9 of 10 osteoarthritis (OA) patient fluids. Upon examining the responses of HSF in culture, OSM stimulated monocyte chemoattractant protein 1 (MCP-1), whereas RANTES secretion (regulated upon activation, normal T expressed and presumably secreted) was not altered by OSM alone. In IL-1alpha-induced cells, OSM costimulation further enhanced MCP-1 release, but inhibited the release of RANTES and IL-8. Other members of the IL-6/LIF family of cytokines did not show these effects. OSM induced a small elevation of MMP-1 production over 2 and 3 days of stimulation (2-fold), and acted significantly to enhance IL-1alpha-induced production of MMP-1 (to 8-fold and 9-fold at 48 and 72 hours, respectively). No effect of OSM was seen on MMP-3 secretion, either alone or in IL-1alpha costimulated cells. CONCLUSION: These results suggest that OSM has potentially important functions in the modulation of chemokine and metalloproteinase production by synovial cells of the joint. PMID- 9416851 TI - Antifibroproliferative effect of tenidap in chronic antigen-induced arthritis. AB - OBJECTIVE: To determine whether tenidap regulates extracellular matrix metabolism in chronic arthritis. METHODS: Antigen arthritis was induced in the knees of 30 rabbits. Animals were distributed into 3 groups: untreated, tenidap-treated, and diclofenac-treated rabbits. Three weeks after disease induction, synovial membranes were extracted and processed for histopathologic examination and detection of type I collagen (CI) and fibronectin (FN) by immunoperoxidase. Simultaneously, we analyzed the in vitro effect of tenidap on healthy synovial cell (SC) proliferation, FN expression and synthesis, and expression of transforming growth factor beta1 (TGFbeta1) messenger RNA. RESULTS: Untreated animals showed synovial lining hyperplasia, cellular infiltration at the sublining, and increased deposition of matrix proteins. These findings were not apparent in tenidap-treated rabbits, where CI and FN had the same distribution as in healthy synovial membranes. In vitro, tenidap inhibited SC proliferation (> or =25 microM) and down-regulated the expression and synthesis of FN in a dose dependent manner (> or =1 microM). This antifibrotic effect was associated with a reduction of TGFbeta1 message. CONCLUSION: Tenidap down-regulates the fibroproliferative changes typical of chronic arthritis, an effect that fits the profile of a disease-modifying agent for rheumatoid arthritis. PMID- 9416852 TI - Effects of recombinant human osteogenic protein 1 on the production of proteoglycan, prostaglandin E2, and interleukin-1 receptor antagonist by human articular chondrocytes cultured in the presence of interleukin-1beta. AB - OBJECTIVE: Recombinant human osteogenic protein 1 (OP-1) is an effective stimulator of human cartilage 35S-proteoglycan synthesis. The present study was conducted to determine whether stimulation of human articular chondrocytes with OP-1 can help overcome interleukin-1beta (IL-1beta)-induced suppression of 35S proteoglycan synthesis. METHODS: Human articular chondrocytes in alginate beads were maintained for 3 days in the absence (control) or presence of IL-1beta at 0.1-100 pg/ml with or without OP-1 at 50 ng/ml, in medium containing 10% fetal bovine serum (FBS). Incorporation of 35S-sulfate into proteoglycans was quantified during the last 4 hours of culture and reported as counts per minute per microg DNA. Release of interleukin-1 receptor antagonist (IL-1Ra) and prostaglandin E2 into the medium was monitored by immunoassay. RESULTS: IL-1beta at 10 pg/ml caused a 60% decrease in 35S-proteoglycan synthesis. This could be blocked by including 500 ng/ml IL-1Ra in the medium. The presence of 50 ng/ml OP 1 in the IL-1beta-containing medium was effective in restoring 35S-proteoglycan synthesis to the level of that found in cultures not treated with IL-1beta. The restorative effects of OP-1 and IL-1Ra were cumulative. The rate of release of prostaglandin E2 and IL-1Ra into the medium was not affected by the presence of OP-1. CONCLUSION: Treatment of human articular chondrocytes with OP-1 cultured in the presence of FBS is effective in overcoming the down-regulation of proteoglycan synthesis induced by low doses of IL-1beta. PMID- 9416853 TI - Histone-specific Th0 and Th1 clones derived from systemic lupus erythematosus patients induce double-stranded DNA antibody production. AB - OBJECTIVE: To investigate whether histone-specific T helper (Th) cells that are able to induce anti-double-stranded DNA (anti-dsDNA) antibodies can be isolated from patients with systemic lupus erythematosus (SLE) and to characterize the cytokine secretion pattern of such Th clones. METHODS: Peripheral blood mononuclear cells from SLE patients and healthy donors were stimulated with autologous apoptotic cell material or purified histones, expanded with interleukin-2 (IL-2), and cloned by limiting dilution. Histone reactivity of clones was examined by histone-specific proliferation and cytokine release. Cytokines were determined by enzyme-linked immunosorbent assay (ELISA) and CTLL-2 bioassay. Induction of anti-dsDNA antibodies was measured in cocultures of autologous B cells and Th clones by ELISA: RESULTS: Numerous histone-specific T cell receptor (TCR) alpha/beta+ Th clones were established from 2 of 3 patients with active SLE and from 1 of 2 healthy individuals. Most Th clones secreted IL 2, interferon-gamma (IFNgamma), and IL-4, whereas some produced predominantly IL 2 and IFNgamma. Th clones that could stimulate the production of anti-dsDNA antibodies were derived from SLE patients and from a healthy individual. CONCLUSION: Th cells specific for histones may play an important role in the pathogenesis of SLE by inducing autoantibodies to dsDNA. Both Th1 and Th2 cytokines may be involved in the pathogenesis of SLE. The presence of histone specific Th cells in a healthy individual indicates the importance of peripheral tolerance for preventing autoimmunity to nuclear antigens. PMID- 9416854 TI - Heterogenous nuclear RNP C1 and C2 core proteins are targets for an autoantibody found in the serum of a patient with systemic sclerosis and psoriatic arthritis. AB - OBJECTIVE: To determine a target recognized by anti-Bh autoantibody, found in the serum of a patient with the unusual coexistence of systemic sclerosis (SSc) and psoriatic arthritis (PsA). METHODS: Antigens recognized by the anti-Bh serum were characterized by indirect immunofluorescence on HeLa cells, by conventional immunoblotting using nuclear extract or partially purified preparation of heterogenous nuclear RNP (hnRNP) proteins, and by 2-dimensional immunoblotting. For the analysis of cross-reactivity and immunofluorescence patterns, autoantibodies were affinity-purified by blot elution and then retested. RESULTS: Comparison of the reactivity of the anti-Bh antibody with the monoclonal antibody 4F4 against both the hnRNP C proteins, together with the determination of biochemical properties of the autoantigens, led to the identification of C1 and C2 core proteins as the targets for the anti-Bh autoantibody. CONCLUSION: Several essential components of the spliceosome are targeted by autoantibodies that are present in the sera of patients with systemic rheumatic diseases. We also found that the hnRNP core proteins C1 and C2 are recognized by the autoantibody present in the serum of a patient with SSc and PsA. C1 and C2 hnRNP proteins should be added to the several intracellular autoantigens recently shown to be cleaved by interleukin-1beta-converting enzyme-like enzymes during apoptosis. PMID- 9416855 TI - Increased phosphate content in complement component C3, fibrinogen, vitronectin, and other plasma proteins in systemic lupus erythematosus: covariation with platelet activation and possible association with thrombosis. AB - OBJECTIVE: To investigate whether extracellular phosphorylation of plasma proteins takes place in vivo in patients with systemic lupus erythematosus (SLE), to determine possible correlations between phosphate levels and clinical and/or laboratory parameters, and to identify individual phosphorylated plasma proteins. METHODS: Sera from SLE patients were analyzed for total amounts of protein-bound phosphate by a colorimetric technique, and for levels of beta-thromboglobulin by radioimmunoassay. In addition, the ability of these sera to activate platelets, resulting in the release of protein kinase, was tested using an assay in which platelet-rich plasma from healthy blood donors was incubated with sera or immune complexes from SLE patients. In this assay, [gamma-32P]ATP was added, and 32P labeled C3 was quantified. Phosphate in individual proteins was detected by Western blot analysis. RESULTS: 32P-labeled, activated platelets were able to phosphorylate exogenously added proteins, without the addition of ATP or cations. Platelet-rich plasma from healthy blood donors became activated by sera or by polyethylene glycol-precipitated immune complexes from patients with SLE, which led to the extracellular phosphorylation of plasma proteins, exemplified in the C3 assay. The phosphate content in plasma proteins was increased in SLE patients with previous thrombosis. The degree of phosphorylation increased up to 3-fold in serial samples obtained from 2 SLE patients during periods of disease exacerbation. Substantial phosphate increases were seen in C3 and fibrinogen. The changes were linked to platelet activation because of the observed covariation with the levels of beta-thromboglobulin. CONCLUSION: In SLE patients, the phosphate content in plasma proteins (including C3 and fibrinogen) increases due to platelet activation. PMID- 9416856 TI - A prospective, multicenter, randomized trial comparing steroids and pulse cyclophosphamide versus steroids and oral cyclophosphamide in the treatment of generalized Wegener's granulomatosis. AB - OBJECTIVE: To investigate the effectiveness and side effects of oral versus pulse cyclophosphamide (CYC) in combination with corticosteroids (CS) in the treatment of systemic Wegener's granulomatosis (WG). METHODS: Patients with newly diagnosed systemic WG were enrolled in a prospective, randomized trial. At the time of diagnosis, prior to randomization, every patient received a daily injection of methylprednisolone for 3 days, followed by daily oral prednisone (1 mg/kg/day) and a 0.7-gm/m2 pulse of CYC. Patients were then randomly assigned to receive either prednisone plus intravenous pulse CYC (group A) or prednisone plus oral CYC (group B) as first-line treatment. CYC was given for at least 1 year and was then progressively tapered and discontinued. RESULTS: Fifty patients were included in the study: 27 in group A and 23 in group B. At 6 months, 24 group A patients (88.9%) were in remission, versus 18 group B patients (78.3%). At the end of the trial, 18 group A patients (66.7%) and 13 group B patients (56.5%) were in remission. In group A, 66.7% of the patients experienced side effects, versus 69.6% in group B. Infectious side effects were significantly more frequent in group B (69.6%) than in group A (40.7%) (P < 0.05). The incidence of Pneumocystis carinii pneumonia was higher in oral CYC-treated patients (30.4%) than in pulse CYC-treated patients (11.1%). Nine group A patients (33.3%) and 10 group B patients (43.5%) died. Actuarial curves showed that relapses were significantly more frequent in group A (59.2%) than in group B (13%) (P = 0.02). CONCLUSION: Our results indicate that pulse CYC is as effective as oral CYC in achieving initial remission of WG and is associated with fewer side effects and lower mortality. However, in the long term, treatment with pulse CYC does not maintain remission or prevent relapses as well as oral CYC. PMID- 9416857 TI - Risk factors for early work disability in systemic lupus erythematosus: results from a multicenter study. AB - OBJECTIVE: To study the risk factors for early work disability in systemic lupus erythematosus (SLE). METHODS: A sample of 159 SLE patients who had been employed at some time since diagnosis was drawn from a multicenter study of outcome in SLE. Disease activity, organ damage, education, income, source of health insurance, and work-related factors were measured in a standardized interview. Work disability was defined by patient self-report of not working because of SLE. The outcome measure was current work status. Seven patients were excluded from the analysis because their choice not to work was unrelated to SLE. RESULTS: An average of 3.4 years after diagnosis, 40% had quit work completely, and job modification was substantial. Univariate analysis (chi-square and t-test) showed that significant predictors of early work disability included having a high school education or less, receiving Medicaid or having no health insurance, having a job which required more physical strength, having an income below poverty level, and having greater disease activity at diagnosis. In multivariate models, significant predictors were education level (P = 0.0004), higher physical demands of the job (P = 0.0028), and higher disease activity at diagnosis (P = 0.0078). Race, sex, cumulative organ damage at diagnosis, and disease duration were not significant. CONCLUSION: Early work disability in SLE is strongly associated with some sociodemographic factors that might be amenable to intervention. PMID- 9416858 TI - A promoter polymorphism of tumor necrosis factor alpha associated with systemic lupus erythematosus in African-Americans. AB - OBJECTIVE: The polymorphic tumor necrosis factor alpha (TNFalpha) gene encodes a cytokine involved in inflammation, angiogenesis, and apoptosis. One polymorphic variant is associated with increased production of TNFalpha. This study examined the frequency of this polymorphic variant in African-American patients with systemic lupus erythematosus (SLE) compared with controls. METHODS: We determined the gene frequency of the polymorphic variant of TNFalpha in an African-American SLE patient population and in a geographically matched African-American control population. RESULTS: The gene frequency of the TNFalpha -308A polymorphism was higher in the African-American SLE population than in the control population. This relationship was independent of major histocompatibility complex DR alleles. CONCLUSION: The TNFalpha -308A polymorphism is associated with an increased risk of SLE in African-Americans. PMID- 9416859 TI - Autoantibodies to human recombinant erythropoietin in patients with systemic lupus erythematosus: correlation with anemia. AB - OBJECTIVE: To investigate the existence of circulating autoantibodies to erythropoietin (EPO) in sera from patients with systemic lupus erythematosus (SLE), and to correlate their presence with anemia and clinical activity. METHODS: Ninety-two consecutive patients with SLE, 80 patients with rheumatoid arthritis, and 42 normal individuals were studied. The patients with SLE were categorized into 3 groups according to hemoglobin (Hgb) level: group A (45 patients with Hgb > 12 gm/dl), group B (26 patients with Hgb 10.1-12 gm/dl), and group C (21 patients with Hgb < or = 10 gm/dl). In all patients with SLE, the disease activity was evaluated using the European Consensus Lupus Activity Measurement scale. Antibodies to EPO were detected using an enzyme-linked immunosorbent assay and purified recombinant human EPO as antigen. The specificity of the method was evaluated with homologous and cross-reactive inhibition assays. RESULTS: Antibodies to EPO were found in 15.2% of the SLE patient sera. The distribution of these antibodies among the 3 groups of SLE patients was as follows: 8.8% (4 of 45) from group A, 15.4% (4 of 26) from group B, and 28.6% (6 of 21) from group C. The prevalence of antibodies to EPO in patients with severe anemia (group C) was statistically significantly higher compared with patients without anemia (chi(2) = 4.31, P < 0.05). Patients with antibodies to EPO had higher disease activity scores (P < 0.005) and lower levels of the C4 component of complement (P < 0.05) compared with patients without antibodies to EPO. CONCLUSION: In this study, the presence of antibodies to EPO in the sera of SLE patients is demonstrated for the first time. The presence of these antibodies is associated with severe anemia and active disease. PMID- 9416860 TI - Circulating plasma levels of nucleosomes in patients with systemic lupus erythematosus: correlation with serum antinucleosome antibody titers and absence of clear association with disease activity. AB - OBJECTIVE: To assess nucleosome plasma levels in patients with systemic lupus erythematosus (SLE) and to study the correlations with serum antinucleosome, anti double-stranded DNA (anti-dsDNA), and antihistone antibody activities, as well as with disease activity (by the SLE Disease Activity Index [SLEDAI]). METHODS: In a cross-sectional study, we assessed 58 SLE patients for their plasma nucleosome levels. Plasma nucleosome levels as well as serum antinucleosome, anti-double stranded DNA, and antihistone antibody activities were assessed by enzyme-linked immunosorbent assay. SLE activity was evaluated using the SLEDAI: RESULTS: The mean (+/-SD) plasma nucleosome concentration in SLE patients was 52 +/- 159 ng/ml (range 5-1,180), and was significantly higher than that of the controls (16 +/- 8.8 ng/ml, range 8-52; P = 0.03). Thirteen of the 58 lupus patients had levels over the range of normal (defined as the control mean + 3 SD, or 42 ng/ml). An inverse correlation was found between nucleosome plasma levels and serum antinucleosome antibody activity in the entire group of SLE patients, those with active disease, and those with inactive disease, respectively. No correlation was found between the SLEDAI and nucleosome plasma concentrations. CONCLUSION: Nucleosome plasma levels may be normal or increased in SLE, and found in patients with active or inactive SLE. Longitudinal studies are needed to further establish whether high levels of circulating nucleosomes may predict the occurrence of an SLE flare. PMID- 9416861 TI - The long-term effect of methotrexate therapy on the liver in patients with juvenile rheumatoid arthritis. AB - OBJECTIVE: To determine if the long-term use of methotrexate (MTX) in juvenile rheumatoid arthritis (JRA) is associated with the development of significant liver fibrosis, and to describe the presence of risk factors for liver fibrosis in patients with JRA. METHODS: Needle biopsies of the liver were performed on a cross-section cohort of 14 patients with JRA who had received a total cumulative dose of MTX that was either > 3,000 mg or > 4,000 mg/1.73 m2 of body surface area. Biopsy samples were independently graded according to the Roenigk Classification Scale by 2 pathologists. The presence of risk factors for MTX hepatotoxicity, especially biochemical abnormalities reflective of liver injury and alcohol consumption, were assessed. RESULTS: Thirteen biopsy samples (93%) were classified as grade I, and 1 (7%) as grade II; none demonstrated significant fibrosis. However, histologic abnormalities were found in 13 biopsy samples (93%). Only 2 patients (14%) consumed more than 1 alcoholic drink per month. Thirteen patients (93%) had biochemical abnormalities while being treated with MTX, but only 5 patients (36%) had at least 1 determination in which the aspartate or alanine aminotransferase elevation was > 3 times the upper limit of normal. CONCLUSION: Long-term use of MTX for JRA does not appear to be associated with the development of significant liver fibrosis. Although nearly all patients had minor histologic changes, no significant clinical consequences were apparent. A prospective study of a larger population will more accurately define the incidence of MTX-related liver fibrosis and appropriate monitoring guidelines in JRA. PMID- 9416862 TI - Psychosocial outcomes and health status of adults who have had juvenile rheumatoid arthritis: a controlled, population-based study. AB - OBJECTIVE: The goal of this study was to evaluate the physical and psychosocial impact of juvenile rheumatoid arthritis (JRA) among a population-based cohort of adults who had the disease during childhood, compared with a control cohort of subjects with no history of JRA. METHODS: The Rochester Epidemiology Project database was used to identify all cases of JRA (based on the American College of Rheumatology [formerly, the American Rheumatism Association] 1977 criteria) among Rochester, Minnesota residents first diagnosed between January 1, 1960 and December 31, 1993. Controls were age- and sex-matched to the cases as of the date of diagnosis of JRA. A pretested postal survey was mailed to all adult cases (whose date of birth was before December 31, 1975) and matched controls from the same population, to obtain information on socioeconomic issues and functional status (using the Health Assessment Questionnaire and the Health Status Questionnaire). The complete medical records of all cases and controls were reviewed to obtain information on demographics and clinical manifestations of JRA. RESULTS: Of the 50 eligible cases, 44 (88%) responded to the survey. There were 102 age- and sex-matched controls (2-3 per case) who responded to the survey. Seventy-three percent of the cases had pauciarticular-onset JRA, 16% had polyarticular-onset JRA, and 11% had systemic-onset JRA. Average followup was 24.7 years and 24.5 years after the index date for cases and controls, respectively. Greater disability (P = 0.0002), more bodily pain (P = 0.0002), increased fatigue (P = 0.0112), poorer health perception (P = 0.0004), and decreased physical functioning (P = 0.0002) were reported by the cases compared with the controls. JRA cases reported significantly lower rates of employment (P = 0.015) and lower levels of exercise (P = 0.0002) than did controls. Level of educational achievement, annual income, health insurance status, and rate of pregnancy and childbirth were similar for both cases and controls. CONCLUSION: Adults who have had JRA during childhood experience long-term physical and psychosocial impairment. PMID- 9416863 TI - Lack of association of HLA-DRB1 genotype with radiologic progression in Japanese patients with early rheumatoid arthritis. AB - OBJECTIVE: To evaluate the role of HLA-DRB1 genotypes in the development and progression of the rheumatoid arthritis (RA) disease process. METHODS: Patients with polyarthritis of < 1 year in duration were consecutively enrolled in the study. Other inclusion criteria were no diagnosis of inflammatory diseases other than RA, and no history of taking disease-modifying antirheumatic drugs or steroids. Patients were evaluated every 4 weeks, and radiographs of the hands/wrists and feet were taken at presentation and 1 year later. HLA-DRB1 genotypes were determined by polymerase chain reaction and restriction fragment length polymorphism methods. RESULTS: We enrolled 198 patients (median disease duration 5.0 months) and 150 controls. The frequency of individuals with DRB1*0405 and *0410 was significantly higher in the patients than in the controls. Homozygous states for DRB1 alleles with the RA-related shared epitope (SE) were associated with increased susceptibility for the development of polyarthritis (odds ratio 3.4, 95% confidence interval 1.5-7.7). None of the DRB1 alleles or SE genotypes correlated with the presence of bone erosion at presentation or 1 year later. CONCLUSION: DRB1 alleles with SEs were associated with the development of polyarthritis but not with early radiographic progression of the disease process. PMID- 9416864 TI - Risk for periodontal disease in patients with longstanding rheumatoid arthritis. AB - OBJECTIVE: To quantify periodontal disease in rheumatoid arthritis (RA) patients and controls, and to correlate the degree of destruction from periodontal disease and from RA. METHODS: Fifty RA patients were matched for age, sex, smoking status, and oral hygiene with 101 controls. Correlations between indices of chronic destruction in periodontal disease (gingival attachment loss) and in RA (Larsen radiographic score) were determined. RESULTS: Patients with longstanding active RA (mean +/- SD 13 +/- 8 years) who were receiving treatment with disease modifying antirheumatic drugs (n = 46), corticosteroids (n = 38), or nonsteroidal antiinflammatory drugs (n = 43) had a higher rate of gingival bleeding (increased by 50%), greater probing depth (increased by 26%), greater attachment loss (increased by 173%), and higher number of missing teeth (increased by 29%) compared with controls. No correlation was found between the Larsen radiographic score and gingival attachment. CONCLUSION: Patients with longstanding active RA have a substantially increased frequency of periodontal disease, including loss of teeth, compared with controls. Antiinflammatory treatment interferes with periodontal disease and might have masked a possible correlation between the indices of chronic destruction in RA and periodontal disease. PMID- 9416866 TI - Effect of age and osteoarthritis on knee proprioception. AB - OBJECTIVE: To test the hypotheses that 1) knee position sense declines with age; 2) patients with osteoarthritis (OA) have worse knee position sense than elderly controls; and 3) knee position sense is correlated with functional status. METHODS: The threshold for detection of knee joint displacement was measured in 30 patients with bilateral knee OA (Kellgren/Lawrence grade > or =2 in both knees), 29 elderly controls (who met clinical and radiographic criteria for exclusion of OA), and 25 young controls. Range of motion, laxity, radiographic severity, and functional status were also assessed. RESULTS: A moderate correlation was found between joint displacement detection threshold and age (r = 0.598 and r = 0.501 for the right knee and the left knee, respectively). The threshold was substantially and significantly different between the OA patients and the elderly controls. Proprioceptive impairment was associated with worse disease-specific functional status. CONCLUSION: Proprioception declines with age, and is further impaired in elderly patients with knee OA. Poor proprioception may contribute to functional impairment in knee OA. PMID- 9416865 TI - Esophageal dysfunction in scleroderma: relationship with disease subsets. AB - OBJECTIVE: To investigate the relationship between esophageal function and the extent of disease in a nonselected group of scleroderma patients, and to study gastric and small bowel motility in a group of scleroderma patients with more severe clinical manifestations. METHODS: Esophageal function in 125 scleroderma patients was investigated by radiologic, endoscopic, manometric, and pH-metric techniques. Ten patients also underwent gastrointestinal (GI) manometric recording, both during fasting and after a standard meal. RESULTS: Radiologic abnormalities of the esophagus were found in 55 of 81 patients (68%) and esophagitis in 45 of 125 (36%). No significant relationship was disclosed between GI symptoms, radiologic abnormalities, esophagitis grade, and the various disease subsets. However, the overall incidence of endoscopic esophagitis (irrespective of the degree) was significantly (P < 0.05) correlated with the patient subgroups, with 100% incidence of esophagitis in those having the more severe cutaneous involvement (type III). Manometric abnormalities were documented in 80% of patients, and pathologic reflux in 78%. The severity of esophageal abnormalities on manometry significantly correlated with the severity of the disease, whereas no correlations were found with pH-metric data. Ninety percent of the 10 female patients undergoing antroduodenal manometry displayed abnormal findings; of these, 60% showed neuropathic, and 30% myopathic, patterns. The latter were recorded in patients with a more severe stage of the disease (type III). CONCLUSION: A direct relationship was observed between scleroderma subsets and the severity of esophageal (and, probably, more distal gut) motor involvement. Since no correlation was found between esophageal symptoms and the severity of manometric abnormalities, manometry should be considered the single most important GI test to document the severity of the "esophageal" disease. Gastric and small bowel manometry may also offer evidence of widespread gut involvement, and provide a rationale for a more targeted therapeutic approach. PMID- 9416867 TI - A case of spontaneous femoral neck fracture associated with multicentric reticulohistiocytosis: oversecretion of interleukin-1beta, interleukin-6, and tumor necrosis factor alpha by affected synovial cells. AB - We describe a case of left femoral neck fracture associated with multicentric reticulohistiocytosis (MR). Biopsy specimens from a skin nodule and from synovial tissue showed histiocytic multinucleated giant cells (MR cells) that are characteristic of MR. A surgical specimen from the resected femoral head revealed that multinucleated giant cells and mononuclear cells invaded the marginal subchondral bone, without evident pannus. These cells also infiltrated into the fracture site, with bone resorption by activated osteoclasts. Immunohistochemical studies of synovium from the left hip joint showed positive staining for interleukin-1beta (IL-1beta), IL-6, and tumor necrosis factor alpha, and abundant cytokine production by cultured synovial cells was demonstrated. These findings suggest that the subchondral invasion and intramedullary infiltration by MR cells caused articular destruction and/or fracture as a result of oversecretion of the cytokines. PMID- 9416868 TI - Regulatory T cell epitope recognized by T cells from labial salivary glands of patients with Sjogren's syndrome. PMID- 9416869 TI - Specialty training and distribution of work effort among US American College of Rheumatology members caring for children with rheumatic disease. PMID- 9416870 TI - Abnormal pain perception in patients with fibromyalgia: comment on the article by Bendtsen et al. PMID- 9416871 TI - Cervical cord compression complicating fractured ossification of the posterior longitudinal ligament: comment on the article by Olivieri et al. PMID- 9416872 TI - Pneumatosis cystoides intestinalis associated with autoantibodies against proliferating cell nuclear antigen: comment on the article by Nojima et al. PMID- 9416873 TI - United Network for Organ Sharing. PMID- 9416874 TI - Early surgery for mitral regurgitation: the advantages of youth. PMID- 9416875 TI - Lowering cholesterol in patients with coronary heart disease: are we ready yet? PMID- 9416876 TI - Tangled up in blue: molecular cardiology in the postmolecular era. PMID- 9416877 TI - Hypercontractility of vascular muscle in atherosclerosis. PMID- 9416878 TI - Baroreflexes and the failing human heart. PMID- 9416879 TI - Heat shock proteins and ischemic injury to the myocardium. PMID- 9416880 TI - Need for a composite risk stratification of patients with unstable coronary syndromes tailored to clinical practice. PMID- 9416881 TI - Individual recognition by heart rate variability of two different autonomic profiles related to posture. AB - BACKGROUND: Power spectrum analysis of heart rate variability (HRV) can estimate the state of sympathovagal balance modulating sinus node activity. In view of the large distribution of spectral variables, a recognition of well-defined physiological conditions has never been attempted on an individual basis. METHODS AND RESULTS: We considered 10 spectral variables extracted from short segments (200 to 500 cardiac cycles) of 350 ECG tracings recorded in normal subjects in both supine and upright positions (700 patterns). The tracings were first ordered consecutively and subsequently assigned alternatively to a training or to a test set (each consisting of 175 cases, providing 350 patterns considered to be independent). A forecasting linear method estimated a normalized activation index (ranging from -1 for supine to +1 for upright) that concentrated the information derived from spectral variables and that identified, in the test set, individual by individual, approximately 84% of corresponding body postures. CONCLUSIONS: The combined use of spectral methodology and forecasting analysis has revealed an information content embedded, per se, in a short series of RR intervals capable of recognizing, individual by individual, two different autonomic profiles related to posture. PMID- 9416882 TI - A genome-based resource for molecular cardiovascular medicine: toward a compendium of cardiovascular genes. AB - BACKGROUND: Large-scale partial sequencing of cDNA libraries to generate expressed sequence tags (ESTs) is an effective means of discovering novel genes and characterizing transcription patterns in different tissues. To catalogue the identities and expression levels of genes in the cardiovascular system, we initiated large-scale sequencing and analysis of human cardiac cDNA libraries. METHODS AND RESULTS: Using automated DNA sequencing, we generated 43,285 ESTs from human heart cDNA libraries. An additional 41,619 ESTs were retrieved from public databases, for a total of 84,904 ESTs representing more than 26 million nucleotides of raw cDNA sequence data from 13 independent cardiovascular system based cDNA libraries. Of these, 55% matched to known genes in the Genbank/EMBL/DDBJ databases, 33% matched only to other ESTs, and 12% did not match to any known sequences (designated cardiovascular system-based ESTs, or CVbESTs). ESTs that matched to known genes were classified according to function, allowing for detection of differences in general transcription patterns between various tissues and developmental stages of the cardiovascular system. In silico Northern analysis of known gene matches identified widely expressed cardiovascular genes as well as genes putatively exhibiting greater tissue specificity or developmental stage specificity. More detailed analysis identified 48 genes potentially overexpressed in cardiac hypertrophy, at least 10 of which were previously documented as differentially expressed. Computer-based chromosomal localizations of 1048 cardiac ESTs were performed to further assist in the search for disease-related genes. CONCLUSIONS: These data represent the most extensive compilation of cardiovascular gene expression information to date. They further demonstrate the untapped potential of genome research for investigating questions related to cardiovascular biology and represent a first generation genome-based resource for molecular cardiovascular medicine. PMID- 9416883 TI - Prognostic influence of increased fibrinogen and C-reactive protein levels in unstable coronary artery disease. FRISC Study Group. Fragmin during Instability in Coronary Artery Disease. AB - BACKGROUND: The prognostic influences of fibrinogen and C-reactive protein levels and their relations to myocardial damage in unstable coronary artery syndromes have not been well described. METHODS AND RESULTS: Fibrinogen and C-reactive protein were determined at inclusion and related to outcome after 5 months in 965 patients with unstable angina or non-Q-wave myocardial infarction randomized to 5 weeks with low-molecular-weight heparin or placebo. The probabilities of death were 1.6%, 4.6%, and 6.9% (P=.005) and the probabilities of death and/or myocardial infarction were 9.3%, 14.2%, and 19.1% (P=.002), respectively, in patients stratified by tertiles of fibrinogen (< 3.38, 3.38 to 3.99, and > or = 4.0 g/L). The probabilities of death were 2.2%, 3.6%, and 7.5% (P=.003) after stratification of patient data by tertiles of C-reactive protein level (< 2, 2 to 10, and > 10 mg/L). In logistic multiple regression analysis, increased fibrinogen levels were independently associated with the incidence of death and/or myocardial infarction (P=.013), and elevated C-reactive protein level was associated with the incidence of death (P=.012). The increased relative risk of subsequent death or myocardial infarction in individuals with an elevated fibrinogen level was consistent in most subgroups evaluated; although significantly so only in patients with signs of myocardial damage. CONCLUSIONS: Increased levels of both fibrinogen and C-reactive protein are associated with a worse outcome in patients with unstable coronary artery disease. The increased risk associated with elevated fibrinogen levels is independent of, and additive to, the prognostic influence of myocardial damage. PMID- 9416884 TI - Cholesterol-lowering therapy in women and elderly patients with myocardial infarction or angina pectoris: findings from the Scandinavian Simvastatin Survival Study (4S) AB - BACKGROUND: The Scandinavian Simvastatin Survival Study (4S) demonstrated pronounced reductions in mortality and major coronary events in a cohort of patients with established coronary heart disease (CHD). The present study provides a detailed, post hoc assessment of the efficacy and safety of simvastatin therapy in the following subgroups of 4S patients: those > or = 65 years of age, those < 65 years of age, women, and men. METHODS AND RESULTS: The 4S cohort of 4444 CHD patients included 827 women and 1021 patients > or = 65 years of age. Total cholesterol at baseline was 5.5 to 8.0 mmol/L with triglycerides < or = 2.5 mmol/L. Patients were randomized to therapy with simvastatin 20 to 40 mg daily or placebo for a median follow-up period of 5.4 years. End points consisted of all-cause and CHD mortality, major coronary events (primarily CHD death and nonfatal myocardial infarction), other acute CHD and atherosclerotic events, hospitalizations for CHD and cardiovascular events, and coronary revascularization procedures. Mean changes in serum lipids were similar in the different subgroups. In patients > or = 65 years of age in the simvastatin group, relative risks (95% confidence intervals) for clinical events were as follows: all-cause mortality, 0.66 (0.48 to 0.90); CHD mortality, 0.57 (0.39 to 0.83); major coronary events, 0.66 (0.52 to 0.84); any atherosclerosis-related event, 0.67 (0.56 to 0.81); and revascularization procedures, 0.59 (0.41 to 0.84). In women, the corresponding figures were 1.16 (0.68 to 1.99); 0.86 (0.42 to 1.74), 0.66 (0.48 to 0.91), 0.71 (0.56 to 0.91), and 0.51 (0.30 to 0.86), respectively. CONCLUSIONS: Cholesterol lowering with simvastatin produced similar reductions in relative risk for major coronary events in women compared with men and in elderly (> or = 65 years of age) compared with younger patients. There were too few female deaths to assess the effects on mortality in women. Because mortality rates increased substantially with age, the absolute risk reduction for both all-cause and CHD mortality in simvastatin-treated subjects was approximately twice as great in the older patients. PMID- 9416885 TI - Circulating adhesion molecules VCAM-1, ICAM-1, and E-selectin in carotid atherosclerosis and incident coronary heart disease cases: the Atherosclerosis Risk In Communities (ARIC) study. AB - BACKGROUND: Recruitment of circulating leukocytes at sites of atherosclerosis is mediated through a family of adhesion molecules. The function of circulating forms of these adhesion molecules remains unknown, but their levels may serve as molecular markers of subclinical coronary heart disease (CHD). METHODS AND RESULTS: To determine the ability of circulating vascular cell adhesion molecule 1 (VCAM-1), endothelial-leukocyte adhesion molecule-1 (E-selectin), and intercellular adhesion molecule-1 (ICAM-1) to serve as molecular markers of atherosclerosis and predictors of incident CHD, we studied 204 patients with incident CHD, 272 patients with carotid artery atherosclerosis (CAA), and 316 control subjects from the large, biracial Atherosclerosis Risk In Communities (ARIC) study. Levels of VCAM-1 were not significantly different among the patients with incident CHD, those with CAA, and control subjects. Higher levels of E-selectin and ICAM-1 were observed for the patients with CHD (means [ng/mL]: E-selectin, 38.4; ICAM-1, 288.7) and those with CAA (E-selectin, 41.5; ICAM-1, 283.6) compared with the control subjects (E-selectin, 32.8; ICAM-1, 244.2), but the distributions were not notably different between the patients with CHD and CAA. Results of logistic regression analyses indicated that the relationship of ICAM-1 and E-selectin with CHD and CAA was independent of other known CHD risk factors and was most pronounced in the highest quartile. The odds of CHD and CAA were 5.53 (95% CI, 2.51-12.21) and 2.64 (95% CI, 1.40-5.01), respectively, for those with levels of ICAM-1 in the highest quartile compared with those in the lowest quartile. Odds of CAA were 2.03 (95% CI, 1.14-3.62) for those with levels of E-selectin in the highest quartile compared with those in the lowest quartile. CONCLUSIONS: These data indicate that plasma levels of ICAM-1 and E-selectin may serve as molecular markers for atherosclerosis and the development of CHD. PMID- 9416886 TI - Reduction of serum cholesterol in postmenopausal women with previous myocardial infarction and cholesterol malabsorption induced by dietary sitostanol ester margarine: women and dietary sitostanol. AB - BACKGROUND: Reduction of serum cholesterol decreases mortality in primary and especially in secondary prevention. We investigated how effectively postmenopausal women with a previous myocardial infarction reduced their serum cholesterol with dietary means by using sitostanol ester rapeseed oil margarine, alone and in combination with statins, and to what extent cholesterol metabolism was affected. METHODS AND RESULTS: The first study group consisted of 22 randomly chosen women with angiographically documented coronary artery disease. Baseline studies on home diet were followed by double-blind, randomized, cross-over studies on margarine without and with sitostanol (3 g/d) ester for 7 weeks in random order. A second group of 10 women on simvastatin consumed sitostanol ester margarine for 12 weeks. Sitostanol ester margarine lowered serum total cholesterol by 13% (P<.05) and LDL cholesterol by 20% (P<.01). Sitostanol ester margarine reduced total cholesterol in all patients, LDL cholesterol <2.6 mmol/L (<100 mg/dL) in 32%, and <3.4 mmol/L (<133 mg/dL) in 73% versus none and 27% during the home diet (P<.01 for both). Combined with simvastatin, sitostanol still reduced total and LDL cholesterol by 11+/-3% and 16+/-5% (P<.01 for both). Sitostanol reduced absorption (-45%), increased fecal elimination (+45% as neutral sterols), and stimulated synthesis (+39%) of cholesterol. High cholestanol and plant sterol (high cholesterol absorption) and low baseline precursor sterol proportions (low cholesterol synthesis) predicted high decreases in serum cholesterol. CONCLUSIONS: Dietary use of sitostanol ester margarine normalizes LDL cholesterol in about one third of women with previous myocardial infarction, especially in those with high baseline absorption and low synthesis of cholesterol, and in combination with statins reduces the needed drug dose. PMID- 9416887 TI - Cyclosporin A inhibits monocyte tissue factor activation in cardiac transplant recipients. AB - BACKGROUND: Fibrin deposition and thrombosis have been implicated in both allograft rejection and vasculopathy after cardiac transplantation. Because monocytes play a pivotal role in the pathophysiology of intravascular coagulation activation through their ability to synthesize tissue factor (TF), we asked (1) whether monocyte TF activation occurs in cardiac transplant recipients and (2) whether monocyte TF expression is affected by treatment with cyclosporin A (CsA). METHODS AND RESULTS: We measured levels of TF activity in peripheral blood mononuclear cells and highly purified monocytes/macrophages from 10 consecutive cardiac transplant recipients and 10 healthy control subjects. TF activity generated by both unstimulated and endotoxin-stimulated cells was significantly higher in transplant recipients than in control subjects (P<.05). Increased monocyte TF expression in transplant recipients was shown to be adversely affected by treatment with CsA: TF induction was markedly reduced by CsA serum concentrations reaching peak CsA drug levels. Inhibition of TF induction in the presence of high CsA blood concentrations was also observed when stimulation of cells was performed with interferon-gamma or interleukin-1beta. As shown by reverse transcription-polymerase chain reaction and electrophoretic mobility shift assay, respectively, treatment with CsA leads to decreased TF mRNA expression and reduced activation of the NF-kappaB transcription factor, which is known to contribute to the induction of the TF promotor in human monocytes. CONCLUSIONS: This study demonstrates that TF activation, occurring in mononuclear cells of cardiac transplant recipients, is inhibited by treatment with CsA. Inhibition of monocyte TF induction by CsA may contribute to its successful use in cardiac transplant medicine and might be useful in managing further settings of vascular pathology also known to involve TF expression and NF-kappaB activation. PMID- 9416888 TI - Effect of the ACE inhibitor lisinopril on mortality in diabetic patients with acute myocardial infarction: data from the GISSI-3 study. AB - BACKGROUND: Mortality of diabetic patients with acute myocardial infarction (MI) remains high despite recent improvement in their management. There is a need to evaluate efficacy and safety of novel treatments of MI in this high-risk population. We evaluated whether treatment with an ACE inhibitor begun within 24 hours from the onset of symptoms is able to decrease mortality and morbidity of diabetic patients with acute MI. METHODS AND RESULTS: A retrospective analysis of the data of the GISSI-3 study in patients with and without a history of diabetes was performed. Patients with suspected acute MI were randomized to treatment with lisinopril (2.5 to 5 up to 10 mg/d) with or without nitroglycerin (5 to 20 microg I.V. then 10 mg/d) begun within 24 hours and continued for 6 weeks. The main end point was mortality at 6 weeks, and the secondary end point was a combined evaluation of mortality and severe left ventricular dysfunction. Information on diabetic status was available for 18,131 patients (approximately 94% of the total population enrolled), of whom 2790 patients had a history of diabetes. Treatment with lisinopril was associated with a decreased 6-week mortality in diabetic patients (8.7% versus 12.4%; OR, 0.68; 95% CI, 0.53 to 0.86; 37+/-12 lives saved per 1000 treated patients), an effect that was significantly (P<.025) higher than that observed in nondiabetic patients. The survival benefit in diabetics was mostly maintained at 6 months despite withdrawal from treatment at 6 weeks (12.9% versus 16.1%; OR, 0.77; 95% CI, 0.62 to 0.95). CONCLUSIONS: Early treatment with the ACE inhibitor lisinopril in diabetic patients with acute MI is associated with a decreased 6-week mortality. This beneficial effect supports a widespread and early use of ACE inhibitors in diabetic patients with acute MI. The burden of mortality plus morbidity for ventricular dysfunction in diabetics remains clinically important and warrants further testing of novel therapeutic approaches. PMID- 9416889 TI - Exaggerated reactivity to mental stress is associated with exercise-induced myocardial ischemia in an asymptomatic high-risk population. AB - BACKGROUND: This study was done to determine whether cardiovascular reactivity to mental stress is associated with exercise-induced occult ischemia in an asymptomatic population at high risk for premature coronary heart disease (CHD). METHODS AND RESULTS: One hundred fifty-two siblings of persons with premature CHD underwent mental stress testing. Exercise thallium tomography and 24-hour Holter monitoring were also performed. Hemodynamic changes were monitored during both stressors. Siblings positive for exercise-induced ischemia were offered cardiac catheterization. During mental stress, siblings with an abnormal exercise ECG and/or thallium scan (n=15) had greater maximal increases in systolic blood pressure (SBP, P=.0004) and diastolic blood pressure (DBP, P=.05) and had greater heart rate variability in the normalized low frequency domain of an analysis of Holter monitor recordings, compared with siblings without exercise-induced ischemia. Coronary arteriography confirmed coronary atherosclerosis in 85% of siblings with exercise-induced ischemia. Regression analyses showed that occult ischemia during exercise was a strong independent predictor of maximal change in SBP and DBP during mental stress. A multivariate logistic model demonstrated that siblings with exercise-induced occult ischemia were 21 times more likely to be "hot" responders (top quartile of change in SBP and DBP) during mental stress. CONCLUSIONS: An exaggerated cardiovascular response to mental stress is associated with exercise-induced myocardial ischemia in persons with preclinical coronary heart disease. PMID- 9416891 TI - Direct intracoronary evidence of collateral steal in humans. AB - BACKGROUND: Coronary steal is defined as a fall in blood flow toward a certain vascular region in favor of another area during arteriolar vasodilatation, ie, a coronary flow velocity reserve (CFVR) <1. The purpose of this study was to determine the frequency of steal in patients with a wide range of collateral supply to a vascular area of interest and to assess whether steal is associated with the amount of collateral flow. METHODS AND RESULTS: One hundred patients 57+/-9 years old with a coronary artery stenosis to be dilated were examined with intracoronary (IC) Doppler guidewires. IC adenosine-induced CFVR<1 obtained distal to the stenosis was defined as steal. An index for collateral flow was determined by positioning the Doppler guidewire in the collateral-dependent vessel distal to the stenosis and measuring the flow velocity time integral during (Vi(occl), cm) and after (Vi(o-occl)) balloon occlusion. Vi(occl)/Vi(o occl) was determined without and with intravenous adenosine (140 microg x kg(-1) x min(-1)). Coronary steal occurred in 10 of 100 patients. Patients with steal showed superior collaterals compared with those without steal: Vi(occl)/Vi(o occl)=0.65+/-0.24 in patients with steal versus 0.29+/-0.18 in those without steal (P=.0001). In all patients with steal, there was a reduction in collateral flow during intravenous adenosine-induced hyperemia, whereas in the majority (70%) of patients without steal, collateral flow increased or remained unchanged during hyperemia. CONCLUSIONS: Coronary steal assessed by intracoronary Doppler flow velocity measurements occurs in 10% of patients with a wide range of coronary collaterals to the vascular area from which blood flow is redistributed. There is a direct association between the presence of steal away from and the amount of collateral flow toward the region under investigation. Collateral flow to the vascular region studied decreases during adenosine-induced hyperemia, which indicates a mechanism of steal via the extensive collaterals. PMID- 9416890 TI - Sphygmomanometrically determined pulse pressure is a powerful independent predictor of recurrent events after myocardial infarction in patients with impaired left ventricular function. SAVE investigators. Survival and Ventricular Enlargement. AB - BACKGROUND: There is increasing evidence of a link between conduit vessel stiffness and cardiovascular events, although the association has never been tested in a large post-myocardial infarction patient population. METHODS AND RESULTS: We evaluated the relationship between baseline pulse pressure, measured by sphygmomanometry 3 to 16 days after myocardial infarction, and subsequent adverse clinical events in the 2231 patients enrolled in the SAVE Trial. Increased pulse pressure was associated with increased age, left ventricular ejection fraction, female sex, history of prior infarction, diabetes, and hypertension and use of digoxin and calcium channel blockers. Over a 42-month period, there were 503 deaths, 422 cardiovascular deaths, and 303 myocardial infarctions. Pulse pressure was significantly related to each of these end points as a univariate predictor. In a multivariate analysis, pulse pressure remained a significant predictor of total mortality (relative risk, 1.08 per 10 mm Hg increment in pulse pressure; 95% CI, 1.00 to 1.17; P<.05) and recurrent myocardial infarction (relative risk, 1.12; 95% CI, 1.01 to 1.23; P<.05) after control for age; left ventricular ejection fraction; mean arterial pressure; sex; treatment arm (captopril or placebo); smoking history; history of prior myocardial infarction, diabetes, or hypertension; and treatment with beta blockers, calcium channel blockers, digoxin, aspirin, or thrombolytic therapy. CONCLUSIONS: These data provide strong evidence for a link between pulse pressure, which is related to conduit vessel stiffness, and subsequent cardiovascular events after myocardial infarction in patients with left ventricular dysfunction. PMID- 9416892 TI - Dipyridamole-induced ischemia as a prognostic marker of future adverse cardiac events in adult patients with hypertrophic cardiomyopathy. Echo Persantine Italian Cooperative (EPIC) Study Group, Subproject Hypertrophic Cardiomyopathy. AB - BACKGROUND: Myocardial ischemia may play a role in the natural history of hypertrophic cardiomyopathy (HCM). To assess the relative prevalence and the prognostic value of dipyridamole-induced ischemia, 79 patients with HCM and without concomitant coronary artery disease (53 men; mean age, 46+/-15 years) underwent a high-dose (up to 0.84 mg/kg over 10 minutes) dipyridamole test with 12-lead ECG and two-dimensional echo monitoring and were followed up for a mean of 6 years. METHODS AND RESULTS: Twenty-nine patients (37%) showed ECG (ie, ST depression > or = 2 mV) signs of myocardial ischemia during dipyridamole test (group 1), whereas 50 (63%) had a negative test (group 2). No patient had transient wall motion abnormalities during the dipyridamole test. During the follow-up, 16 events (ie, left ventricular or atrial enlargement, unstable angina, syncope, atrial fibrillation, and bundle-branch block) occurred in 29 patients in group 1 and 5 in 50 patients in group 2 (55% versus 10%, P<.001). Patients with a positive dipyridamole test showed worse 72-month event-free survival rates compared with patients with a negative test (36.2% versus 84.2%, P<.001). A forward stepwise event-free survival analysis identified dipyridamole test positivity by ECG criteria (chi2=19.7, P=.0001), rest gradient (chi2=11.3, P=.0008), and age (chi2=4.1; P=.0413) as independent and additive predictors of subsequent events. CONCLUSIONS: ECG signs of myocardial ischemia elicited by dipyridamole are frequent in patients with HCM and identify patients at higher risk of cardiac events, suggesting a potentially important pathogenetic role of inducible myocardial ischemia in determining adverse cardiac events in these patients. PMID- 9416893 TI - Diastolic ventricular interaction: a possible mechanism for abnormal vascular responses during volume unloading in heart failure. AB - BACKGROUND: Baroreflex dysfunction is common in chronic heart failure and contributes to the associated sympathoexcitation. Baroreceptor activity normally decreases during volume unloading, causing an increase in sympathetic outflow and resulting in forearm vasoconstriction. Some heart failure patients develop attenuated vasoconstriction or paradoxical vasodilation. The mechanism for this is unknown. We have recently demonstrated diastolic ventricular interaction in some patients with chronic heart failure as evidenced by increases in left ventricular (LV) end-diastolic volume in association with decreases in right ventricular (RV) volume during volume unloading. We reasoned that such an increase in LV volume, by increasing LV mechanoreceptor activity, would decrease sympathetic outflow and could therefore explain the abnormal vascular responses seen in such patients. METHODS AND RESULTS: We assessed changes in forearm vascular resistance (FVR) during application of -20 and -30 mm Hg lower-body negative pressure (LBNP) in 24 patients with chronic heart failure and 16 control subjects. Changes in LV and RV end-diastolic volumes were assessed during -30 mm Hg LBNP in all heart failure patients. Diastolic ventricular interaction was demonstrated in 12 patients as evidenced by increases in LV end-diastolic volume in association with decreases in RV end-diastolic volume during LBNP. Changes in FVR during LBNP (-20 and -30 mm Hg) were markedly attenuated in these 12 patients (-1.6+/-11.2 and -0.9+/-12.5 U) compared with both the remaining patients (11.9+/ 10.0 and 17.0+/-12.3 U) and the control subjects (16.5+/-9.5 and 23.1+/-13.9 U) (P<.01 for both comparisons at each level of LBNP). FVR decreased in 5 of these 12 patients during -30 mm Hg LBNP, a response seen in none of the remaining patients (P=.01). CONCLUSIONS: Diastolic ventricular interaction in patients with chronic heart failure is associated with attenuated forearm vasoconstriction or paradoxical vasodilation during LBNP. This may explain the apparent derangement in baroreflex control of sympathetic outflow during acute volume unloading in heart failure. PMID- 9416894 TI - Late left ventricular function after surgery for children with chronic symptomatic mitral regurgitation. AB - BACKGROUND: The use of quantitative echocardiography has been emphasized in optimizing timing of surgery in adult patients with mitral regurgitation to avoid irreversible left ventricular dysfunction. In contrast, surgery for infants and children is often delayed until the appearance of severe symptoms because of the patient's size and anticoagulation requirements and the possible need for early reoperation. The purpose of this study was to determine long-term ventricular function after mitral valve surgery in symptomatic children and to analyze risk factors for adverse outcome. METHODS AND RESULTS: Thirty-three patients (0.5 to 19 years old) operated on for mitral regurgitation as a single hemodynamically significant lesion were studied. All but 3 had medically refractory symptoms. One patient died during surgery, and 32 were followed for 0.3 to 17.1 years (mean, 4.5 years). The mean preoperative left ventricular shortening fraction was 0.38+/ 0.09. Successful mitral valvuloplasty or replacement was documented by long-term normalization of end-diastolic dimensions. Early postoperative shortening fraction was significantly reduced (0.28+/-0.1, P<.01), but it improved to 0.40+/ 0.07 (P<.01) on late follow-up, at which time only 1 patient had ventricular dysfunction. Preoperative shortening fractions did not correlate well with early or late postoperative values (r=.18 and r=.31, respectively). Seven of 32 surviving patients had preoperative shortening fractions <0.33 (mean, 0.26+/ 0.05) and 25 >0.33 (mean, 0.39+/-0.08). Analysis of these subgroups showed no significant differences between the groups in early or late postoperative function. Duration of mitral insufficiency appeared to be associated with the development of atrial arrhythmias. CONCLUSIONS: Late left ventricular function normalizes in children after surgical correction of mitral insufficiency. In contrast to adults, delay of surgery in children with significant mitral regurgitation until the onset of severe symptoms does not increase the risk for long-term ventricular dysfunction, although late atrial arrhythmias are more likely to be encountered. PMID- 9416895 TI - Tomographic three-dimensional echocardiographic determination of chamber size and systolic function in patients with left ventricular aneurysm: comparison to magnetic resonance imaging, cineventriculography, and two-dimensional echocardiography. AB - BACKGROUND: Two-dimensional (2D) echocardiographic approaches based on geometric assumptions face the greatest limitations and inaccuracies in patients with left ventricular (LV) aneurysms. Three-dimensional (3D) echocardiographic techniques can potentially overcome these limitations; to date, however, although tested in experimental models of aneurysms, they have not been applied to a series of patients with such distortion. The purpose of this study was therefore to validate the clinical application of tomographic 3D echocardiography (3DE) by the routine transthoracic approach to determine LV chamber size and systolic function without geometric assumptions in patients with LV aneurysms. METHODS AND RESULTS: In 23 patients with chronic stable LV aneurysms, LV end-systolic and end diastolic volumes (LVEDV, LVESV) and ejection fraction (LVEF) by tomographic 3DE were compared with results from 3D magnetic resonance tomography (3DMRT) as an independent reference as well as with the conventional techniques of single plane and biplane 2D echocardiography and biplane cineventriculography. Dynamic 3DE image data sets were obtained from a transthoracic apical view with the use of a rotating probe with acquisition gated to control for ECG and respiration (Echoscan, TomTec). Volumes were calculated from the 3D data sets by summating the volumes of multiple parallel disks. 3DE results correlated and agreed well with those by 3DMRT, with better correlation and agreement than provided by other techniques for LVEDV (3DE: r=.97, SEE=14.7 mL, SD of differences from 3DMRT=14.5 mL; other techniques: r=.84 to .93, SEE=30.7 to 41.6 mL [P<.001 versus 3DE by F test], SD of differences=31.5 to 40.7 mL [P<.001 versus 3DE by F test]). The same also pertained to LVESV (3DE: r=.97, SEE=12.4 mL, SD of differences=12.9 mL; other techniques: r=.81 to .90, SEE=24.7 to 37.2 mL [P<.001], SD of differences=27.6 to 36.8 mL [P<.005]) and LVEF (3DE: r=.74, SEE=5.6%, SD of differences=6.7%; other techniques: r=.14 to .59, SEE=9.5% to 10.1% [P<.01], SD of differences=9.5% to 12.6% [P<.05]). Compared with 3DMRT, 3DE was less time consuming and patient discomfort was less. CONCLUSIONS: Tomographic 3DE is an accurate noninvasive technique for calculating LV volumes and systolic function in patients with LV aneurysm. Unlike current 2D methods, tomographic 3DE requires no geometric assumptions that limit accuracy. PMID- 9416896 TI - Antiarrhythmic actions of intravenous ibutilide compared with procainamide during human atrial flutter and fibrillation: electrophysiological determinants of enhanced conversion efficacy. AB - BACKGROUND: The selective class III antiarrhythmic agent ibutilide prolongs action potential duration and terminates atrial flutter (AFL) and fibrillation (AF), but the mechanism of its antiarrhythmic efficacy in humans has not been fully characterized. This study compared the antiarrhythmic effects of ibutilide with the class IA agent procainamide in humans during AFL and AF. Antiarrhythmic drug actions and electrophysiological characteristics of AFL and AF that enhanced pharmacological termination were investigated. METHODS AND RESULTS: Right atrial monophasic action potentials were recorded during 148 episodes of AFL (n=89) or AF (n=59) in 136 patients treated with intravenous ibutilide (n=73) or placebo (n=22) as participants in randomized, double-blinded comparative studies or intravenous procainamide (n=53) in a concurrent open-label study. The conversion rates in AFL with ibutilide, procainamide, and placebo were 64% (29 of 45 patients), 0% (0 of 33), and 0% (0 of 11), respectively, whereas in AF the rates were 32% (9 of 28), 5% (1 of 20), and 0% (0 of 11), respectively. In AFL, ibutilide increased atrial monophasic action potential duration (MAPD) more (30% versus 18%, P<.001) and prolonged atrial cycle length (CL) less (16% versus 26%, P<.001) than procainamide. Ibutilide shortened and procainamide prolonged action potential diastolic interval during AFL (-12% versus 51%, P<.001). Ibutilide increased MAPD/CL ratio, whereas procainamide tended to decrease this ratio (13% versus -6%, P<.01). In AF, ibutilide and procainamide induced similar increases in atrial CL (48% versus 45%), but ibutilide induced a greater increase in MAPD (52% versus 37%, P<.05). Independent electrophysiological predictors of pharmacological arrhythmia termination were increase in MAPD/CL ratio (P=.005) in AFL and longer baseline mean MAPD (P=.011) in AF. Termination of AFL with ibutilide was characterized by significant increases in beat-to-beat atrial CL, MAPD, and diastolic interval variability. Ibutilide was significantly more effective in converting AF when the mean atrial CL was > or = 160 ms (64% versus 0%, P<.001) or MAPD was > or = 125 ms (57% versus 0%, P=.002) at baseline. CONCLUSIONS: Enhanced conversion efficacy of ibutilide compared with procainamide in AFL is correlated with a relatively greater prolongation of atrial MAPD than atrial CL, and termination of AFL by ibutilide is characterized by oscillations in atrial CL and MAPD. Conversion of AF by ibutilide is enhanced by a longer baseline mean atrial CL or MAPD. PMID- 9416897 TI - Ventricular tachycardia in valvular heart disease: facilitation of sustained bundle-branch reentry by valve surgery. AB - BACKGROUND: The clinical characteristics of sustained monomorphic ventricular tachycardia (SMVT), when it develops after valve surgery, have not been described. METHODS AND RESULTS: Between 1985 and 1996, 31 patients (30 men and 1 woman) who had undergone valve surgery were found to have inducible SMVT. Nine patients (29%) had sustained VT due to bundle-branch reentry (BBR) (group 1). Four of these patients had normal left ventricular function, and VT with a right bundle-branch morphology was inducible in 4 patients. Group 2 included 20 patients with inducible myocardial (ie, non-BBR) VT. Coronary artery disease was present in 15 group 2 patients (75%) due to atherosclerotic (n=12) and nonatherosclerotic (n=3) causes. Two patients had both inducible sustained BBR and myocardial VT (group 3). Sustained BBR VT occurred significantly earlier after valve surgery (median, 10 days) than the onset of postoperative myocardial VT (median, 72 months; P<.005). CONCLUSIONS: Myocardial VT was the most common type of inducible SMVT in patients with valvular heart disease. The majority of these patients had underlying coronary artery disease and significant left ventricular dysfunction. However, in almost one third of the patients, sustained BBR VT was the only type of inducible SMVT. This type of VT was facilitated by the valve procedure occurring within 4 weeks after surgery in most patients. In these patients, left ventricular function was relatively well preserved, and the right bundle-branch block type of BBR was frequently induced. Because a curative therapy can be offered to these patients (ie, bundle-branch ablation), BBR should be seriously considered as the mechanism of VT in patients with valvular heart disease, particularly if the arrhythmia occurs soon after valve surgery. PMID- 9416898 TI - Mapping of ventricular repolarization potentials in patients with arrhythmogenic right ventricular dysplasia: principal component analysis of the ST-T waves. AB - BACKGROUND: Nonuniform recovery of ventricular excitability has been demonstrated to facilitate the reentry circuits leading to the development of ventricular tachyarrhythmias. This can also occur in arrhythmogenic right ventricular dysplasia (ARVD). In fact, in patients with ARVD, abnormalities of ventricular repolarization are often observed on 12-lead ECGs, but their predictive value for the occurrence of malignant arrhythmias is yet to be established. Because body surface potential mapping has been proved to be useful for the detection of heterogeneities in ventricular recovery even though they are not revealed by conventional 12-lead ECGs, we attempted to analyze repolarization potentials on the entire chest surface to find abnormalities that can be predictive of ventricular arrhythmias. METHODS AND RESULTS: Body-surface potential maps were recorded from 62 anterior and posterior thoracic leads in 22 patients affected by ARVD, 9 with episodes of sustained ventricular tachycardias (VT) and 13 without. Thirty-five healthy subjects were also studied as control subjects. The 62 chest ECGs were simultaneously recorded, digitally converted at a rate of 2000 Hz, and stored on a hard disk of a body-surface mapping computer system. In each subject, the QRST integral map was obtained by calculating at each lead point the algebraic sum of all instantaneous potentials, from the QRS onset to the T-wave end, multiplied by the sampling interval. In most ARVD patients, we observed a larger-than-normal area of negative values on the right anterior thorax. This abnormal pattern could be explained by a delayed repolarization of the right ventricle. Nevertheless, it was not related to the occurrence of VT in our patient population. To detect minor heterogeneities of ventricular repolarization, the principal component analysis was applied to the 62 ST-T waves recorded in each subject. We assumed that a low value of the first or of the first three components (components 1, 2, and 3) indicates a greater-than-normal variety of the ST-T waves, a likely expression of a more complex recovery process. The mean values of the first three components were not significantly different in ARVD patients and control subjects. Nevertheless, considering the two subsets of patients with and without VT, the values of component 1, components 1 + 2, and component 1 + 2 + 3 were significantly lower in the group of ARVD patients with VT. Values of component 1 < 69% (equal to 1 SD below the mean value for control subjects) were found in 6 of 9 VT patients and in 1 patient without VT (sensitivity, 67%; specificity, 92%). A low value of component 1 was the only variable significantly associated with the occurrence of VT. CONCLUSIONS: Principal component analysis provides a better quantitative assessment of the complexity of repolarization than other ECG measurements. When applied to ARVD patients, principal component analysis of the ST-T waves recorded from the entire chest surface revealed abnormalities not detected by conventional ECG that can be considered indexes of arrhythmia vulnerability. PMID- 9416899 TI - Summary measures of the insulin resistance syndrome are adverse among Mexican American versus non-Hispanic white children: the Corpus Christi Child Heart Study. AB - BACKGROUND: Mexican-American (MA) adults are known to have a greater burden of diabetes and insulin resistance than non-Hispanic white (NHW) people. In this report, we examined data obtained from MA and NHW third-grade children for evidence of a pattern consistent with the insulin resistance syndrome. In addition, we developed two summary measures characterizing insulin resistance syndrome to compare measures of this syndrome among our population. METHODS AND RESULTS: Data regarding fasting insulin, triglycerides, HDL cholesterol, systolic blood pressure, and body mass index (BMI) were available for 403 third-grade children. Median levels of insulin and glucose were significantly higher in MA boys and girls than in NHW boys and girls. Risk factors characterizing insulin resistance, including levels of insulin, triglycerides, systolic blood pressure, HDL cholesterol, and BMI were categorized as above or below the total population median. MA children were more likely than NHW children to have three or more adverse risk factors (55% versus 37%). When risk factors were converted to Z scores, and the five Z scores were summed for each individual, MA boys and girls had higher mean scores than NHW boys and girls (means for boys, 0.65 versus 0.97, P<.0001; girls, 0.52 versus -0.30, P<.04). Principal components analysis was used to create a summary score or index representing the insulin resistance syndrome. This summary score was significantly higher among MA boys and girls than NHW boys and girls (means for boys, 0.34 versus -0.72, P<.0001; girls, 0.35 versus -0.04, P=.056). CONCLUSIONS: Our results support the hypothesis that MA children exhibit a greater degree of the insulin resistance syndrome than NHW children, especially among boys. We conclude that some of the factors responsible for the increased risk of NIDDM seen among MA adults are demonstrable in childhood. PMID- 9416900 TI - Relationship between repeated measures of hemodynamics, muscle sympathetic nerve activity, and their spectral oscillations. AB - BACKGROUND: We determined the intraclass correlation coefficients (ICC) of repeated measures of the mean levels and variability of RR and muscle sympathetic nerve activity (MSNA) in 7 normal subjects. We examined whether spontaneous fluctuations in RR and MSNA over repeated measurements were mirrored by changes in spectral components of RR and MSNA. METHODS AND RESULTS: Twenty-minute recordings of respiration, RR, blood pressure (BP), and MSNA were performed at day 1, 1 week, 1 month, and 3 months and divided into two 10-minute periods for the analysis of short-term reliability. Comparison between these recordings also determined the long-term reliability. Linear regressions examined the relationship between changes in these measurements and changes in spectral components of RR and MSNA. All analyses were carried out blinded to subject and session. Short-term ICC of RR, BP, MSNA and of the variabilities of RR and MSNA (in % of total power) ranged between .98 and .70 and indicated a good short-term reliability. The long-term reliability of RR variability was comparable to MSNA variability (range of ICC, .34 to .52). Spontaneous decreases in RR during the repeated recordings were accompanied by increases in sympathetic drive, as evidenced by increases in the ratio of low-frequency to high-frequency variability (LF/HF ratio) of RR interval (r=-.43, P<.01) and by increases in MSNA (r= -.36, P=.01). The changes in the LF/HF ratio of RR were mirrored by parallel changes in the LF/HF ratio of MSNA (r=+.30, P<.05). Spontaneous decreases in BP were accompanied by increases in the LF/HF ratio of MSNA (r=-.52; P=.0001). CONCLUSIONS: Heart rate, MSNA, and their variability are stable in the short term, but less so over the long term. Spontaneous changes in repeated measurements of RR interval and blood pressure over the long term are accompanied by parallel changes in the normalized spectral components of RR and MSNA variability. Thus even over an extended period, there is a synchrony between changes in absolute cardiovascular measures and changes in their spectral components. PMID- 9416901 TI - Remodeling and neointimal formation in the carotid artery of normal and P selectin-deficient mice. AB - BACKGROUND: Inflammatory reactions such as leukocyte activation with platelet adherence and release of inflammatory mediators occur after percutaneous transluminal coronary angioplasty and may play a role in restenosis. Vascular remodeling with neointimal formation was studied in normal C57Bl/J6 and P selectin-deficient mice. METHODS AND RESULTS: The left common carotid artery was ligated just proximal to the carotid bifurcation. Four weeks later, left carotids and contralateral controls were snap-frozen. Computer-aided morphometry was performed to measure ratios of neointimal to medial area (NI/M) in 10 sections per animal as a measure of the thickness of the neointimal lesion. For normal mice, NI/M was 1.13+/-0.2 (n=20), whereas NI/M was reduced by 76% to 0.27+/-0.1 (n= 19) in P-selectin knockout mice. Vascular constriction (as measured by the length of external elastic lamina) was the same in both groups, but the circumference of the lumen in knockout mice was 26% larger. Also, normal and P selectin-deficient mice were killed at 3 and 7 days after ligation (n=6 for each group per time point). Histological staining and immunostaining for CD45 showed no inflammatory cell presence in P-selectin knockout mice. However, in normal mice, leukocyte infiltration was observed in the adventitia, media, and developing neointima. Also, P-selectin immunostaining was observed in media and developing neointima of normal mice. CONCLUSIONS: These data suggest that P selectin is involved in processes leading to cell migration and proliferation associated with vascular remodeling, presumably by mediating leukocyte recruitment and the interaction between platelets and leukocytes. PMID- 9416902 TI - Small heat shock proteins and protection against ischemic injury in cardiac myocytes. AB - BACKGROUND: Overexpression of the inducible hsp70 protects against ischemic cardiac damage. However, it is unclear whether the small heat shock proteins hsp27 and alphaB-crystallin protect against ischemic injury. METHODS AND RESULTS: Our aim was to examine whether the overexpression of hsp27 and alphaB-crystallin in neonatal and adult rat cardiomyocytes would protect against ischemic injury. Recombinant adenovirus expressing hsp27 or alphaB-crystallin under the control of the cytomegalovirus promoter was used to infect cardiac myocytes at high efficiency as assessed by immunostaining. Overexpression was confirmed by Western blot analysis. Cardiomyocytes were subjected to simulated ischemic stress, and survival was estimated through assessment of lactate dehydrogenase and creatine phosphokinase release. The hsp27 overexpression decreased lactate dehydrogenase release by 45+/-7.5% in adult cardiomyocytes but had no effect in the neonatal cells. In contrast, alphaB-crystallin overexpression was associated with a decrease in cytosolic enzyme release in both adult (29+/-6.6%) and neonatal (32+/ 5.4%) cardiomyocytes. Decreased endogenous hsp25 with an antisense adenovirus produced a 29+/-9.9% increase in damage with simulated ischemia. Overexpression of the inducible hsp70 in adult cardiomyocytes was associated with a 34+/-4.6% decrease in lactate dehydrogenase release and is in line with our previous results in neonatal cardiomyocytes. CONCLUSIONS: The increased expression of hsp27 and alphaB-crystallin through an adenovirus vector system protects against ischemic injury in adult cardiomyocytes. Likewise, the overexpression of alphaB crystallin protects against ischemic damage in neonatal cardiomyocytes. Decreasing the high levels of endogenous hsp25 present in neonatal cardiomyocytes renders them more susceptible to damage caused by simulated ischemia. PMID- 9416903 TI - Effects of adenovirus-mediated human apo A-I gene transfer on neointima formation after endothelial denudation in apo E-deficient mice. AB - BACKGROUND: Inactivation of apolipoprotein (apo) E genes in mice markedly increases beta-VLDL levels and accelerates progression of complex atherosclerotic lesions. The present study investigated (1) the effect of apo E deficiency (apo E /-) on neointima formation after endothelial denudation; and (2) the effect of increased HDL, induced by adenovirus-mediated transfer of a human apo A-I gene, on neointima formation. METHODS AND RESULTS: Guidewire-induced abrasion of the endothelium of the common carotid artery did not produce neointima formation within 18 days after injury in C57BL/6J mice (n=12) but was associated with an intima/media ratio of 0.82+/-0.25 in age-matched C57BL/6J apo E-/- mice (n=12). Neointima consisted primarily of smooth muscle alpha-actin positive cells. Injection in C57BL/6J apo E-/- mice of 2x10(9) (n=5) or 4x10(9) (n=7) plaque forming units (p.f.u.) of a recombinant human apo A-I adenovirus 3 days before injury resulted in an increase of HDL cholesterol from 36+/-5 to 75+/-3 mg/dL (P<.05) and to 96+/-13 mg/dL (P<.05), respectively, and of the HDL cholesterol/non-HDL cholesterol ratio from 0.063+/-0.003 to 0.15+/-0.01 (P<.05) and to 0.16+/-0.015 (P<.05), respectively. Intima/media ratio decreased to 0.28+/ 0.06 (P=NS versus C57BL/6J apo E-/- mice) with 2x10(9) p.f.u. of apo A-I virus and to 0.03+/-0.01 with 4x10(9) p.f.u. (P<.01 versus C57BL/6J apo E-/- mice). Injection of 4x10(9) p.f.u. of RR5 (n=7) or tissue plasminogen activator (t-PA) control virus (n=6) did not result in a significant alteration of HDL cholesterol (44+/-11 and 26+/-4 mg/dL, respectively) nor in a reduction of intima/media ratio (0.81+/-0.35 and 0.86+/-0.23, respectively). CONCLUSIONS: Apo E deficiency is associated with increased neointima formation after endothelial denudation. Gene transfer of apo A-I increases HDL cholesterol and significantly reduces neointima formation, which suggests a direct vascular protective effect of HDL. PMID- 9416904 TI - Enhanced myosin light chain phosphorylations as a central mechanism for coronary artery spasm in a swine model with interleukin-1beta. AB - BACKGROUND: Although coronary artery spasm plays an important role in a wide variety of ischemic heart diseases, the intracellular mechanism for the spasm remains to be clarified. We examined the role of myosin light chain (MLC) phosphorylations, a key mechanism for contraction of vascular smooth muscle, in our swine model with interleukin-1beta (IL-1beta). METHODS AND RESULTS: IL-1beta was applied chronically to the porcine coronary arteries from the adventitia to induce an inflammatory/proliferative lesion. Two weeks after the operation, intracoronary serotonin repeatedly induced coronary hyperconstrictions at the IL 1beta-treated site both in vivo and in vitro, which were markedly inhibited by fasudil, an inhibitor of protein kinases, including protein kinase C and MLC kinase. Western blot analysis showed that during serotonin-induced contractions, MLC monophosphorylation was significantly increased and sustained in the spastic segment compared with the control segment, whereas MLC diphosphorylation was noted only in the spastic segment. A significant correlation was noted between the serotonin-induced contractions and MLC phosphorylations. Both types of MLC phosphorylation were markedly inhibited by fasudil. In addition, MLC diphosphorylation was never induced by a simple endothelium removal in the normal coronary artery, whereas enhanced MLC phosphorylations in the spastic segment were noted regardless of the presence or absence of the endothelium. CONCLUSIONS: These results indicate that enhanced MLC phosphorylations in the vascular smooth muscle play a central role in the pathogenesis of coronary spasm in our swine model. PMID- 9416905 TI - Assisted ventilation during 'bystander' CPR in a swine acute myocardial infarction model does not improve outcome. AB - BACKGROUND: Mouth-to-mouth rescue breathing is a barrier to the performance of bystander cardiopulmonary resuscitation (CPR). We evaluated the need for assisted ventilation during simulated single-rescuer bystander CPR in a swine myocardial infarction model of prehospital cardiac arrest. METHODS AND RESULTS: Steel cylinders were placed in the mid left anterior descending coronary arteries of 43 swine. Two minutes after ventricular fibrillation, animals were randomly assigned to 10 minutes of hand-bag-valve ventilation with 17% oxygen and 4% carbon dioxide plus chest compressions (CC+V), chest compressions only (CC), or no CPR (control group). Standard advanced life support was then provided. Animals successfully resuscitated received 1 hour of intensive care support and were observed for 24 hours. Five of 14 CC animals, 3 of 15 CC+V animals, and 1 of 14 controls survived for 24 hours (CC versus controls, P=.07). Myocardial oxygen delivery and consumption were greater among surviving animals than nonsurvivors but did not differ between CC and CC+V animals. CONCLUSIONS: In this acute myocardial infarction model of prehospital single-rescuer bystander CPR, assisted ventilation did not improve outcome. PMID- 9416906 TI - Halothane protects cardiomyocytes against reoxygenation-induced hypercontracture. AB - BACKGROUND: Resupply of oxygen to the myocardium after extended periods of ischemia or hypoxia can rapidly aggravate the already existing injury by provoking hypercontracture of cardiomyocytes (acute reperfusion injury). Previous studies indicated that halothane can protect ischemic-reperfused myocardium. The aim of the present study was to analyze on the cellular level the mechanism by which halothane may protect against reoxygenation-induced hypercontracture. METHODS AND RESULTS: To simulate ischemia-reperfusion, isolated adult rat cardiomyocytes were incubated at pH 6.4 under anoxia and reoxygenated at pH 7.4 in the presence or absence of 0.4 mmol/L halothane. Reoxygenation was started when intracellular Ca2+ (measured with fura 2) had increased to > or = 10(-5) mol/L and pHi (BCECF) had decreased to 6.5. Development of hypercontracture was determined microscopically. In the control group, reoxygenation provoked oscillations of cytosolic Ca2+ (72+/-9 per minute at fourth minute of reoxygenation) accompanied by development of hypercontracture (to 65+/-3% of end ischemic cell length). When halothane was added on reoxygenation, Ca2+ oscillations were markedly reduced (4+/-2 per minute, P<.001) and hypercontracture was virtually abolished (90+/-4% of end-ischemic cell length, P<.001). Halothane did not influence the recovery of pHi during reoxygenation. Similar effects on Ca2+ oscillations and hypercontracture were observed when ryanodine (3 micromol/L), an inhibitor of the sarcoplasmic reticulum Ca2+ release, or cyclopiazonic acid (10 micromol/L), an inhibitor of the sarcoplasmic reticulum Ca2+ pump, were applied instead of halothane. CONCLUSIONS: Halothane protects cardiomyocytes against reoxygenation-induced hypercontracture by preventing oscillations of intracellular Ca2+ during the early phase of reoxygenation. PMID- 9416907 TI - Hydroxychloroquine reverses thrombogenic properties of antiphospholipid antibodies in mice. AB - BACKGROUND: Previous studies have demonstrated that human monoclonal and polyclonal anticardiolipin antibodies have thrombogenic properties in vivo. Using such a model in which these antibodies have been shown to increase both the size of an induced thrombus and the duration of time in which such a clot lasts, we investigated whether hydroxychloroquine alters the dynamics of such thrombus formation. METHODS AND RESULTS: Three groups of nine mice were injected with purified immunoglobulin G (IgG) from a patient with the antiphospholipid syndrome (IgG-APS) and then fed with hydroxychloroquine at various doses (100, 6, and 3 mg/kg body wt). Three control groups of mice were also studied, including mice injected with IgG-APS and then fed with placebo, as well as two other groups injected with IgG from normal human serum and fed either hydroxychloroquine or placebo. A standardized thrombogenic injury was subsequently induced in the femoral vein of each mouse and the area (size) of thrombus measured as well as the total period of time that thrombus was present. Mice treated with hydroxychloroquine and IgG-APS showed significantly smaller thrombi that persisted for a shorter period of time compared with animals treated with IgG-APS and placebo. CONCLUSIONS: Hydroxychloroquine significantly diminished both thrombus size and total time of thrombus formation in mice previously injected with IgG-APS. PMID- 9416908 TI - Aqueous oxygen: a highly O2-supersaturated infusate for regional correction of hypoxemia and production of hyperoxemia. AB - BACKGROUND: High levels of hyperoxemia may have utility in the treatment of regional tissue ischemia, but current methods for its implementation are impractical. A catheter-based method for infusion of O2, dissolved in a crystalloid solution at extremely high concentrations, ie, 1 to 3 mL O2/g (aqueous oxygen [AO]), into blood without bubble nucleation was recently developed for the potential hyperoxemic treatment of regional tissue ischemia. METHODS AND RESULTS: To test the hypotheses that hypoxemia is correctable and that hyperoxemia can be produced locally by AO infusion, normal saline equilibrated with O2 at 3 MPa (30 bar; 1 mL O2/g) was delivered into arterial blood in two different animal models. In 15 New Zealand White rabbits with systemic hypoxemia, AO was infused into the midabdominal aorta at 1 g/min. Mean distal arterial PO2 increased to 236+/-113 and 593+/-114 mm Hg on 1-hour periods of air and O2 breathing, respectively, from a baseline of 70+/-10 mm Hg (P<.01). In contrast, infusion of ordinary normal saline in a control group (n=7) had no effect on arterial PO2. No differences between groups (P>.05) in temporal changes in blood counts and chemistries were identified. In 10 dogs, low coronary blood flow in the circumflex artery was delivered with a roller pump through the central channel of an occluding balloon catheter. Hypoxemic, normoxemic, and AO induced hyperoxemic blood perfusates (mean PO2, 52+/-4, 111+/-22, and 504+/-72 mm Hg, respectively) were infused for 3-minute periods in a randomized sequence. Short-axis two-dimensional echocardiography demonstrated a significant decrease (P<.05) in left ventricular ejection fraction compared with baseline physiological values with low-flow hypoxemic and normoxemic perfusion but not with low-flow hyperoxemic perfusion. CONCLUSIONS: Intra-arterial AO infusion was effective in these models for regional correction of hypoxemia and production of hyperoxemia. PMID- 9416909 TI - Electrophysiological mechanism of the characteristic electrocardiographic morphology of torsade de pointes tachyarrhythmias in the long-QT syndrome: detailed analysis of ventricular tridimensional activation patterns. AB - BACKGROUND: The long-QT syndrome (LQTS) is an electrophysiological (EP) entity characterized by prolongation of cardiac repolarization and the occurrence of polymorphic ventricular tachyarrhythmias (VTs), sometimes with a twisting QRS morphology, better known as torsade de pointes (TdP). In the present study, detailed analysis of ventricular tridimensional activation patterns during nonsustained TdP VT was performed to provide an EP mechanism of the periodic transition in QRS axis. METHODS AND RESULTS: The studies were conducted with the anthopleurin-A canine model of LQTS. Tridimensional isochronal maps of ventricular activation were constructed from 256 bipolar electrograms obtained from the use of 64 plunge needle electrodes. In 26 episodes of nonsustained TdP VT, detailed activation maps could be accurately constructed during QRS-axis transitions in surface ECGs. The initial beat of all VTs consistently arose as a subendocardial focal activity, whereas subsequent beats were due to reentrant excitation in the form of rotating scrolls. The VT ended when reentrant excitation was terminated. In 22 of 26 episodes, the transition in QRS axis coincided with the transient bifurcation of a predominantly single rotating scroll into two simultaneous scrolls involving both the right ventricle and left ventricle separately. The common mechanism for initiation or termination of bifurcation was the development of functional conduction block between the anterior or posterior right ventricle free wall and the ventricular septum. In 4 of 26 episodes, a fast polymorphic VT, with an apparent shift in QRS axis, was due to a predominantly single localized circuit that varied its location and orientation from beat to beat, with the majority of ventricular myocardium being activated in a centrifugal pattern. CONCLUSIONS: The study provides for the first time an EP mechanism for the characteristic periodic transition of the QRS axis during TdP VT in the LQTS. PMID- 9416910 TI - Additional lead improves defibrillation efficacy with an abdominal 'hot can' electrode system. AB - BACKGROUND: Although the left prepectoral site is preferred for "hot can" placement, this site is unavailable in some patients. We evaluated the influence of electrode location on defibrillation thresholds with alternative hot can and transvenous lead configurations. METHODS AND RESULTS: Three interrelated studies were performed. In group 1, the importance of hot can location was investigated by pairing a right ventricular lead to five different hot can placement sites in seven pigs. The defibrillation energies for right pectoral, left pectoral, left subaxillary, and right and left abdominal hot can sites were 20.3+/-2.7,* 15.9+/ 3.8, 14.9+/-2.5, 32.0+/-3.4,* and 30.0+/-3.4 J,* respectively (*P<.005 versus left pectoral and left subaxillary sites). In group 2, the value of a three electrode configuration with an abdominal hot can placement was investigated by adding a subclavian vein lead to the pectoral or abdominal hot can configurations in seven pigs. The defibrillation energies for left pectoral and abdominal sites were 18.6+/-4.2 and 29.0+/-5.8 J (P=.0001), respectively. The addition of a right or left subclavian vein lead with an abdominal hot can reduced the threshold to 19.3+/-4.2* or 18.8+/-3.2,* respectively (*P=.0001 versus abdominal site). In group 3, the contribution of the abdominal hot can electrode to the three electrode configuration was tested by a comparison with two purely transvenous two-electrode configurations in six pigs. The defibrillation energy (19.9+/-3.2 J) for the abdominal hot can with a subclavian vein lead was lower than the transvenous lead configurations with a subclavian vein (29.0+/-2.5 J, P=.0001) or a superior vena cava lead (30.7+/-3.7 J, P=.0001). The right ventricular lead was the sole cathode during the first phase of the biphasic shock in all experiments. CONCLUSIONS: Defibrillation energy depends on the hot can placement site. The addition of a subclavian vein lead with an abdominal hot can improves defibrillation efficacy to the level of the pectoral placement and is better than a purely transvenous lead configuration. PMID- 9416911 TI - Effect of altering filling pattern on diastolic pressure-volume curve. AB - BACKGROUND: The early-to-late ventricular filling ratio (E:A) is widely used to index diastolic function. While filling patterns reflect diastolic properties, they can also modulate chamber pressures due to myocardial viscoelasticity. We hypothesized that such feedback can potentially temper effects of delayed relaxation and/or volume loading on diastolic pressures. METHODS AND RESULTS: Six isolated blood-perfused canine left ventricles were studied with ejection and filling controlled by an intracavitary volume servo-pump. Diastolic filling was determined by a simulated atrial pressure source that was either constant or varied to yield dual-phase filling at a specified E:A ratio. E:A ratio was randomly set to 3:1, 1:3, or 1:1, and data were recorded at each ratio at three different preloads. With principally early filling (E:A=3:1), diastolic pressure rise from viscosity increased in proportion with the relaxation time constant (r=.91, P<.0001). However, this dependence was lost as E:A ratio declined (eg, P=.63 for E:A 1:3). Furthermore, E:A=3:1 yielded 37% to 50% lower end-diastolic pressures at similar volumes (versus E:A=1:3) as initial viscous forces decayed. Offsetting early and late filling effects led to little net change in mean diastolic pressure independent of E:A ratio or preload. CONCLUSIONS: Diastolic filling pattern itself influences chamber pressures early and late in diastole due to viscoelasticity, with larger net effects on end-diastolic pressure. Since E:A ratio normally falls with delayed relaxation but rises with higher preload or reduced compliance, the present results suggest that changes in filling pattern may modulate direct effects of such factors on elevating diastolic pressure. PMID- 9416912 TI - Regulation of hepatic vascular volume: contributions from active and passive mechanisms during catecholamine and sodium nitroprusside infusion. AB - BACKGROUND: It is unclear how the liver contributes to regulation of cardiac filling. The aims of this study were to establish an animal model to quantify hepatic vascular capacitance and to determine the mechanisms whereby catecholamines and sodium nitroprusside modify hepatic blood volume. METHODS AND RESULTS: In 8 anesthetized pigs we measured hepatic and systemic pressures and flows. Liver vascular volume was measured by sonomicrometry calibrated against integrated hepatic inflow during outflow occlusion. Pressure-volume (P-V) curves were constructed during outflow occlusion. Sonomicrometry accurately reflected hepatic blood volume (r=.99+/-.001), and hepatic P-V curves were highly reproducible. Norepinephrine (0.3 and 0.7 microg x kg body weight (bwt)(-1) min( 1) intraportally) significantly reduced hepatic blood volume by 3.3+/-1 and 4.3+/ 1 mL x kg bwt(-1), respectively. Nitroprusside (8 and 18 microg x kg bwt(-1) x min(-1) intraportally) increased hepatic blood volume by 1.1+/-0.2 and 1.9+/-0.3 mL x kg bwt(-1), respectively. Norepinephrine and nitroprusside parallel shifted the hepatic P-V curves, indicating reduced and increased unstressed blood volume, respectively. These curve shifts accounted for more than 90% of the respective blood volume changes. Compliance was unchanged. Phenylephrine but not isoprenaline yielded similar results as norepinephrine. CONCLUSIONS: The pig model used in this study, accurately quantified hepatic capacitance. Alpha adrenergic stimulation decreased and nitroprusside increased capacitance by changing unstressed blood volume. These changes in capacitance correspond to expulsion of 300 mL and pooling of 130 mL of blood, respectively, in a 70-kg individual, reflecting that the liver is not only a passive blood reservoir but can respond actively and vigorously to pharmacological interventions. PMID- 9416913 TI - Cholesterol management in theory and practice. AB - The preponderance of evidence confirms the importance of aggressive lipid modification in patients at risk for coronary heart disease (CHD). However, data suggest that this information is underimplemented in the clinical setting, even in patients with existing CHD, in whom the greatest benefit of such treatment has been shown. The fact that many practitioners do not pursue a proven treatment strategy in patients who qualify must be redressed through education and reinforcement of existing recommendations. In the present review, the current clinical and mechanistic understanding of the benefit of aggressive lipid management is summarized, with a focus on the clinical implications of recent findings. These include growing public awareness of cholesterol as a modifiable CHD risk factor, recommendations for earlier and more aggressive intervention in patients with existing disease, and discussion of the cost-effectiveness of lipid regulating therapy. Despite the secular trend of declining CHD morbidity and mortality rates in recent years, CHD remains the leading cause of death in both men and women in the United States. It is imperative to prevent any reduction in public focus on primary and secondary prevention. PMID- 9416914 TI - Images in cardiovascular medicine. Aspergillus mitral endocarditis. PMID- 9416915 TI - Images in cardiovascular medicine. Situs inversus with complete transposition in the fetus: diagnostic antenatal sequential segmental analysis. PMID- 9416916 TI - Neurogenesis in the mammalian neostriatum and nucleus accumbens: parvalbumin immunoreactive GABAergic interneurons. AB - We study the neurogenesis of a distinct subclass of rat striatum gamma aminobutyric acid (GABA)ergic interneurons marked by the calcium-binding protein parvalbumin (PV). Timed pregnant rats are given an intraperitoneal injection of bromodeoxyuridine (BrdU), a marker of cell proliferation, on designated days between embryonic day (E) 11 and E22. Birthdate of PV neurons is determined in the adult neostriatum and nucleus accumbens by using a BrdU-PV double-labeling immunohistochemical technique. PV-immunoreactive interneurons of the neostriatum show maximum birthrates (>10% double-labeling) between E14-E17, whereas PV immunoreactive interneurons of the nucleus accumbens show maximum double-labeling between E16-E19. In the neostriatum, caudal PV-immunoreactive neurons are born before those at rostral levels, and lateral PV-immunoreactive neurons become postmitotic before medial neurons. In the postcommissural striatum, ventral PV immunoreactive neurons become postmitotic before dorsal neurons. In the precommissural striatum, ventral neurons are born before dorsal neurons laterally, but a dorsoventral gradient is seen medially. At corresponding coronal levels, PV-immunoreactive neurons of the nucleus accumbens are born shortly after PV neurons of the neostriatum. Analysis of BrdU labeling intensity in the nucleus accumbens shows that medium spiny projection neurons of the shell become postmitotic before neurons of the core. Similarly, PV-immunoreactive interneurons of the nucleus accumbens shell are born before PV interneurons of the core. Compared with cholinergic interneurons of the neostriatum, PV-immunoreactive interneurons are born later, but neurogenetic gradients are similar. The period of striatum PV interneuron genesis encompasses the period for somatostatin interneurons, although the latter neurons do not show neurogenetic gradients, possibly due to heterogeneous subtypes. Consideration of basal telencephalon neurogenesis suggests that subpopulations of striatum interneurons may share common neurogenetic features with phenotypically similar populations in the basal forebrain, with final morphology and connectivity depending on local cues provided by the host environment. PMID- 9416917 TI - Sensory neuroanatomy of a skin-penetrating nematode parasite Strongyloides stercoralis. II. Labial and cephalic neurons. AB - Host recognition, contact, and skin-penetration by Strongyloides stercoralis infective larvae are crucially important behavioral functions mediating transition from free-living to parasitic life. The sensilla of the worm's anterior tip presumably play an important role in these processes. Besides the main chemosensilla, the amphids, which are of central importance, the larva has 16 putative mechanosensilla. There are six inner labial sensilla: two dorsal, two ventral, and two lateral. The two dorsal and ventral pairs are each innervated by two neurons, whereas each lateral sensillum is singly innervated. The six outer labial and four cephalic sensilla are all singly innervated. All of these have the characteristics of mechanoreceptors: they are closed to the external environment, and closely associated with the overlying cuticle. Distally, their dendritic processes contain granular material and associated microtubules. With two exceptions, the relevant neuronal cell bodies lie in lateral ganglia adjacent to the nerve ring, their positions remarkably similar to those of their homologues in the free-living nematode, Caenorhabditis elegans. Cell bodies of two neuronal pairs, one of two dorsal inner labial neurons and one of two ventral inner labial neurons per side, are however, found far anterior to the remaining cell bodies. All labial and cephalic sensilla are apparently mechanoreceptors, complementing the well-developed chemosensilla. Presumably infective larvae require touch and stretch receptors, not only to initiate skin penetration by finding irregularities as points of access, but also to bore through tissue to reach their ultimate enteral destination. PMID- 9416918 TI - Differential distribution of inositol 1,4,5-triphosphate receptors in the rat olfactory bulb. AB - The inositol 1,4,5-triphosphate receptor (IP3R) regulates the release of calcium from intracellular stores. In the present study, the distribution of IP3R in the rat main olfactory bulb was determined by immunohistochemistry. Immunofluorescence was used to double label for IP3R and for gamma-aminobutyric acid (GABA) or for projection neurons, which were retrogradely labeled following dextran injection into the lateral olfactory tract (LOT). The expression profile of IP3R changes dramatically during development. In the glomerular layer of adults, many juxtaglomerular neurons are IP3R immunoreactive [IP3R(+)]; the majority of these cells are also GABA immunoreactive [GABA(+)]. Scattered sparsely throughout the external plexiform layer are small numbers of IP3R(+) neurons, a small number of which are LOT-projecting tufted cells. Significant numbers of IP3R(+) neurons are in the granule cell layer; however, most of these cells are GABA(-). The vast majority of mitral cells contain little or no IP3R immunoreactivity. These findings indicate that, in the olfactory bulb of adult rats, IP3R is preferentially localized in specific classes of intrinsic neurons and that it is rarely expressed in projection neurons. In contrast, during the first postnatal week, the receptor is detected almost exclusively in mitral cells. Expression of IP3R in subclasses of intrinsic neurons begins during the second and third weeks, concomitant with a decrease in immunostaining of mitral cells. Adult patterns of IP3R immunostaining are apparent by the fourth postnatal week. These observations raise the possibility that the expression of IP3R in specific classes of neurons during development is activity dependent. PMID- 9416919 TI - Synaptic inputs to identified color-coded amacrine and ganglion cells in the turtle retina. AB - Previous studies have proposed models of the specific synaptic circuitry responsible for color processing in the turtle retina. To determine the accuracy of these models of the circuits underlying color opponency in the inner retina of the turtle (Pseudemys scripta), we have studied the physiology, morphology, and synaptic connectivity of identified amacrine and ganglion cells. These cells were first characterized electrophysiologically and were then stained with horseradish peroxidase. Postembedding electron immunocytochemistry for gamma-aminobutyric acid (GABA) and glycine was used to reveal the neurochemical identity of their synaptic inputs. The red-ON/green, blue-OFF small-field ganglion cell, classified as G24, branched primarily in strata S1, S4, and S5 of the inner plexiform layer (IPL). Ganglion cell G24 showed a complex receptive field organized into a red-ON center surrounded by an inhibitory region, which, in turn, was surrounded by a second excitatory region. Only the center responses were color opponent. The red OFF/green, blue-ON large-field, stellate amacrine cell, classified as A23b, stratified exclusively in stratum S2, near the S2/S3 border. The color-coded center was surrounded by a luminosity, red-sensitive surround. Synaptic input to G24 and A23b was dominated by amacrine cells (89% and 87%, respectively). G24 received significant input from amacrine cell profiles with GABA (13% of total) as well as glycine (11% of total) immunoreactivity, mostly in the proximal stratum S5 of the IPL (64% and 67% of the total GABA- and glycine-immunoreactive input, respectively). Bipolar cell synaptic input was also found predominantly in S4 and S5 (89%). In contrast, we found no glycine-immunoreactive input to A23b, and the density of the GABA-immunoreactive amacrine cell synaptic input revealed a central (15%) to peripheral (3%) gradient within the dendritic tree. The results of the present study support the previous models of the synaptic circuitry responsible for color-opponent signal processing in the inner retina of the turtle. PMID- 9416920 TI - Somatotopical organisation within the climbing fibre projection to the paramedian lobule and copula pyramidis of the rat cerebellum. AB - The climbing fibre projection to the paramedian lobule (lobule VII) and the copula pyramidis (lobule VIII) in the posterior lobe of the rat cerebellum was investigated in pentobarbitone-anaesthetised animals. Percutaneous electrical stimulation generated climbing fibre field potentials on the cerebellar surface that indicated a somatotopical organisation into five distinct cortical areas. Each area was identified by the site(s) of peripheral stimulation that evoked the largest response within that area, and from medial to lateral the cortex was subdivided as follows [principal site(s) of stimulation in parentheses with corresponding range of onset latencies]: in the paramedian lobule, area 1 (contralateral face, 17-18 ms), area 2 (ipsilateral forelimb, 10-15 ms) and area 3 (ipsi- and contralateral forelimbs, 16-26 ms and 15-30 ms, respectively); and in the copula pyramidis, area 4 (tail, 17-21 ms) and area 5 (ipsilateral hindlimb, 13-19 ms). In additional retrograde tracer experiments, small volumes of fluorescent-tagged beads were injected into each of the different areas and the location of retrogradely labelled olive cells was mapped. By comparison with other species, the results indicate that in the paramedian lobule area 1 corresponds to zone A2; area 2 corresponds, at least in part, to medial C1; and area 3 corresponds to C2; in addition, farther laterally, a C3 zone may be present. In the copula pyramidis, areas 4 and 5 correspond to subzones of medial C1. Overall, the results support the view that a general principle of cerebellar cortical organisation is a division into parasagittal zones, each characterised by its somatotopically organised climbing fibre input that arises from a specific, rostrocaudally oriented column of cells within the inferior olivary nucleus. PMID- 9416921 TI - Patterns of gamma-aminobutyric acid and glycine immunoreactivities reflect structural and functional differences of the cat lateral lemniscal nuclei. AB - The three nuclei of the cat lateral lemniscus (dorsal, intermediate, and ventral) were distinguished by their immunoreactivities for the putative inhibitory transmitters, gamma-aminobutyric acid (GABA) and glycine. Each nucleus had a distinct pattern of somatic and perisomatic labeling. The dorsal nucleus contained mostly GABA-immunoreactive neurons (85%), with moderate numbers of GABA and glycine-immunoreactive puncta along their somata. The remaining neurons were nonimmunoreactive (15%). The intermediate nucleus contained mostly nonimmunoreactive neurons (82%), and these had numerous glycine-immunoreactive and few GABA-immunoreactive perisomatic puncta. The remaining neurons were immunoreactive for GABA only (10%), glycine only (2%), or both (6%). The ventral nucleus contained mostly glycine-immunoreactive neurons (81%), and about half of these were also GABA-immunoreactive. The remaining neurons were either nonimmunoreactive (8%) or GABA-immunoreactive only (11%). Neurons in the ventral nucleus had fewer immunoreactive perisomatic puncta than neurons in either the dorsal or the intermediate nuclei. These differences in neuronal immunoreactivity and in the relative abundance of GABA-and glycine-immunoreactive perisomatic puncta among the three nuclei of the lateral lemniscus support connectional and electrophysiological evidence that each nucleus has a different functional role in auditory processing. In particular, this study demonstrates that the intermediate nucleus of the cat is cytochemically distinct from the dorsal and ventral nuclei in terms of the somatic and perisomatic immunoreactivity of its neurons for these two important inhibitory transmitters and may provide novel inputs to the inferior colliculus. PMID- 9416922 TI - Sex difference and steroidal stimulation of galanin immunoreactivity in the ferret's dorsal preoptic area/anterior hypothalamus. AB - A sexually dimorphic male nucleus (MN) is seen in Nissl-stained sections from the dorsal preoptic area/anterior hypothalamus (dPOA/AH) of male, but not female, ferrets. We used immunohistochemical methods to determine whether particular neuropeptides are found in cells of the MN. A sexually dimorphic cluster of galanin-immunoreactive (IR) cells was found in the dPOA/AH of ferrets killed either on embryonic day (E) 38 or in adulthood. Significantly more galanin-IR cells were distributed in the MN and in other subregions of the dPOA/AH of intact breeding males than estrous females. The density of galanin-IR cells in the dPOA/AH was significantly reduced in adult males by castration and restored to the level of intact breeding males by daily injections of testosterone propionate (TP) for 5 weeks. The same TP treatment failed to augment the density of galanin IR cells in the dPOA/AH of adult, ovariectomized females. Computer-assisted image analysis and grid-crossing analysis showed that the area and the number of galanin-IR fibers in the dPOA/AH were significantly greater in adult females than in males, regardless of subjects' concurrent steroidal condition. A cluster of galanin-IR cells was present in the dPOA/AH of males, but not females, killed on either E34 or E38. Administration of TP between E28 and E37 significantly increased the density of galanin-IR cells in the dPOA/AH of females killed on E38, up to the level seen in control males. The results suggest that the capacity of cells located in the dPOA/AH to express galanin after adult steroid exposure is sexually differentiated by the fetal action of testosterone, or its metabolite, estradiol, in males. PMID- 9416923 TI - Regulation of the polysialylated form of the neural cell adhesion molecule in the developing striatum: effects of cortical lesions. AB - Early in development, the polysialylated form of the neural cell adhesion molecule (PSA-NCAM) is expressed by growth cones, neuronal processes, and neuronal cell bodies. In rat striatum, PSA-NCAM expression becomes progressively restricted to pre- and postsynaptic membranes and is undetectable by postnatal day 25 (P25), i.e., after corticostriatal synaptogenesis. This study examined the effects of cortical lesions performed on P14, when the corticostriatal projection is already primarily unilateral and cortical inputs have not yet formed asymmetric synapses on striatal neurons. Rats were killed on P25, and PSA-NCAM expression was examined by immunoblotting and immunohistochemistry with light and electron microscopy. In contrast to the case in controls, PSA-NCAM expression was maintained in the striatum of lesioned pups. Ultrastructural studies showed that PSA-NCAM was present 1) in growth cone-like structures and neuronal processes and 2) in striatal neurons. Together with the presence of growth cones, the observation that the number of asymmetric synapses was unchanged in the denervated striatum suggests that axonal sprouting occurred in response to the lesion. This was confirmed by axonal labeling in the denervated striatum after injection of Fluoro-Ruby in the contralateral cortex. The data indicate that P14 cortical lesions affect PSA-NCAM expression in the developing striatum 1) by inducing a robust axonal plasticity resulting in the presence of immature presynaptic elements that contain PSA-NCAM and 2) by delaying the loss of PSA NCAM expression in striatal neurons, suggesting that the lesion affects the time course of striatal maturation. PMID- 9416924 TI - Somatodendritic and axonal anatomy of intracellularly labeled serotonergic neurons in the rat medulla. AB - A knowledge of the anatomy of medullary serotonergic cells is critical to understanding local and brainstem circuits in which these cells participate. Serotonergic neurons (n = 16) were identified, as previously described (Mason [1997] J. Neurophysiol. 77:1087-1098) by their slow and steady background discharge in halothane anesthetized rats. Neurons were then intracellularly labeled with Neurobiotin and visualized with 3,3'diaminobenzidine. The validity of the physiological identification of serotonergic cells was confirmed by processing two neurons that were physiologically characterized as serotonergic for serotonin immunoreactivity; both tested cells contained immunoreactive serotonin. The dendrites and axon of each labeled cell were reconstructed by using a three-dimensional computerized system. Somata were small or medium in size and had fusiform, triangular, or multipolar shapes. The dendritic arbor was constricted with most dendrites extending for less than 500 microm from the soma. All labeled axons projected caudally and travelled in the ventrolateral medulla, either dorsal or ventral to the lateral reticular nucleus. Most cells had collaterals and/or dense axonal swellings in the nucleus reticularis gigantocellularis, nucleus reticularis magnocellularis, raphe magnus, and the ventrolateral medulla. Non-local collaterals and swellings were also observed in the nucleus reticularis gigantocellularis and in the ventrolateral medulla at all medullary levels. The results demonstrate that 1) the dendrites of serotonergic cells are restricted to raphe magnus and the ventral part of nucleus reticularis magnocellularis; and 2) serotonergic cells project to medullary nuclei that contain bulbospinal cells which project to dorsal, intermediate, and ventral horns. Serotonergic cell projections to brainstem sites may mediate the integration of sensory, autonomic, and motor modulation at the brainstem level. PMID- 9416925 TI - Ageing has a differential effect on nitric oxide synthase-containing and catecholaminergic amacrine cells in the human and rat retina. AB - In this study, we assessed the effects of normal ageing on the number, distribution, and somal area of nicotinamide adenine dinucleotide phosphate diaphorase (NADPHd)-positive (NADPHd+) and tyrosine hydroxylase-immunoreactive (TH-IR) amacrine cells in human and rat retina. By using a double-labelling immunohistochemical technique, we have shown that these two enzymes are located in separate amacrine cell populations in the human retina. In normal human retinas from organ donors, we have shown that there was no change in the number, somal area, or retinal distribution of NADPHd+ neurons over an age range of 19-89 years. In contrast, there was a significant decrease (P < 0.05) of 52% in the total number of TH-IR neurons in the group aged 65-89 years compared with the group aged 19-64 years. CA1 and CA2 TH-IR neurons were reduced by 44% and 55%, respectively. In young (3 months) and old (2 years) rats, the number of NADPHd+ neurons did not decrease with ageing, but the number of TH-IR neurons was significantly reduced by 21% (P < 0.05). In a companion study on monkey retina, we have shown that a postmortem delay of 12.5 hours between death and fixation results in a decrease of 33% in the number of both NADPHd+ and TH-IR neurons in the retina compared with the number in retinas fixed immediately after death. The findings of this study on the two subsets of amacrine cells, therefore, are likely to demonstrate the consequences of ageing in the retina and might contribute to visual impairment in the elderly. PMID- 9416927 TI - Utility of stress single-photon emission computed tomography (SPECT) perfusion imaging in predicting outcome after coronary artery bypass grafting. AB - Previous studies have examined the predictors of outcome in medically treated patients with coronary artery disease (CAD). There is limited information on predictors of outcome after coronary artery bypass grafting (CABG). This study examined the predictors of outcome of 255 patients with CAD, at a mean time of 5 years after CABG for angina pectoris. The 255 patients underwent coronary angiography and stress single-photon emission computed tomography (SPECT) myocardial perfusion imaging after CABG. During a mean follow-up of 41 +/- 28 months after stress testing, there were 34 hard events (24 cardiac deaths and 10 nonfatal myocardial infarctions). The hemodynamics during stress testing, and age and gender were not predictors of events. The SPECT variables of multivessel perfusion abnormality, perfusion deficit size, and increased lung thallium uptake were predictors of death and total events by uni- and multivariate survival analysis. There were 14 events in 45 patients (31%) with multivessel abnormality and increased lung thallium uptake, 14 events in 101 patients (14%) with either multivessel abnormality or increased lung uptake, and 6 events in 109 patients (6%) with neither of these 2 variables (p = 0.0001). The annual mortality and total event rates were 7.5% and 9.5% with both variables, 3.4% and 4.3% with either variable, and 0.6% and 1.7% with neither of the variables (p = 0.01). Thus, stress SPECT perfusion imaging is useful to stratify patients after CABG into low, intermediate, and high risk groups for future cardiac events. PMID- 9416926 TI - Topographical distribution of neurons containing endothelin type A receptor in the rat brain. AB - Endothelin (ET) was originally identified as a vasoactive peptide biosynthesized in vascular endothelial cells. Because ET has also been found in the brain as a neuropeptide, it has been thought to belong to the group of brain-vascular peptide hormones. To date, type A and type B receptors for ET have been found. To elucidate the topographic distribution of type A receptor (ET-AR) in the brain, we raised a specific antibody to the C-terminal (64 amino acids) peptide of rat ET-AR and immunostained rat brain sections with this antibody. Immunoreactivity for ET-AR was detected in neuronal cell bodies and also in the many proximal and some distal parts of their fibers. Nerve cell bodies containing strong ET-AR immunoreactivity were distributed in the lateral part of the reticular formation, the nucleus of the solitary tract and its surrounding area, the dorsal midline area and medial longitudinal fasciculus, the subependymal layer of the fourth ventricular roof, the caudolateral area of the pontine tegmentum, the locus coeruleus, the rostral pontine area of the lateral reticular formation, the retrorubral area, the substantia nigra, the ventral tegmental area, the periventricular region lateral to the rostral mesencephalic aqueduct and caudal third ventricle, the arcuate hypothalamic nucleus, the caudomedial area of the zona incerta, the periventricular hypothalamic nucleus, the parvocellular portion of the paraventricular hypothalamic nucleus, and the periglomerular region of the olfactory bulb. In addition, the Purkinje cells of the cerebellar cortex, the nerve cells in the mesencephalic trigeminal nucleus, and the magnocellular neurons of the supraoptic and paraventricular hypothalamic nuclei showed weak immunoreactivity. The distribution of highly ET-AR-immunoreactive neurons is quite similar to that ofcatecholamine neurons. PMID- 9416928 TI - Clinical and prognostic significance of atrial fibrillation in acute myocardial infarction. AB - The clinical significance of the time of onset of atrial fibrillation (AF) was investigated in patients with acute myocardial infarction (AMI). Among 1,039 patients with AMI, 100 (9.6%) had AF. These patients were divided into 3 groups: AF group 1 (n = 45), who developed AF within 24 hours of the onset of AMI; AF group 2 (n = 41), who developed AF >24 hours after the onset of AMI; and AF group 3 (n = 14), who developed AF before the onset of AMI. The infarct-related lesion was most frequent (67%) in the proximal right coronary artery in AF group 1 (p <0.01). Right atrial pressure was most significantly increased in AF group 1. The left atrial dimension and pulmonary arterial wedge pressure were most significantly increased, and left ventricular ejection fraction was most significantly decreased in AF group 2. In the acute phase, the frequencies of heart failure, cardiogenic shock, and in-hospital mortality were higher for all 3 AF groups than the sinus group (p <0.01). The long-term survival rate was significantly lower in AF group 1 and AF group 2 than in the sinus group. AF was an independent predictor of cardiac death in both AF group 1 (odds ratio 2.5; 95% confidence interval 1.2 to 5.0; p = 0.0012) and AF group 2 (odds ratio 3.7; 95% confidence interval 1.8 to 7.5; p = 0.0005), but not in AF group 3. The onset time of AF appears to be a useful parameter for evaluating the cardiac status and prognosis of patients with AMI. PMID- 9416929 TI - Occurrence of sustained increase in QT dispersion following exercise in patients with residual myocardial ischemia after healing of anterior wall myocardial infarction. AB - Our objective was to evaluate the effect of exercise on QT dispersion over the next 3 hours, as seen on a standard 12-lead electrocardiogram in patients with healed myocardial infarction with or without residual ischemia. We measured QT and QTc dispersion before, immediately after, and 1 and 2 hours after symptom limited, dynamic treadmill exercise tests in 28 patients with healed anterior wall myocardial infarction with (group I, n = 18) and without (group II, n = 10) residual ischemia. The same protocol was followed in 5 group I patients after successful performance of coronary angioplasty. QT and QTc dispersion did not change immediately after exercise in group II. These parameters increased in group I (QT dispersion at rest [mean +/- SD] 57 +/- 22 ms, and after exercise 87 +/- 27 ms; QTc dispersion at rest 62 +/- 25 ms, and after exercise 114 +/- 36 ms). The increases in QT and QTc dispersion were sustained for at least 2 hours. After a successful coronary angioplasty in 5 patients, these parameters no longer increased with exercise. Thus, QT dispersion increased for at least 2 hours after exercise in patients who had residual ischemia after healing of myocardial infarction. Data obtained in 5 of these patients after coronary angioplasty support the idea that residual ischemia plays a key role in the sustained increase in QT dispersion after exercise. PMID- 9416930 TI - Attenuation of neutrophil and endothelial activation by intravenous morphine in patients with acute myocardial infarction. AB - To investigate the effects of morphine on neutrophil and endothelial activation, we measured serum levels of intercellular adhesion molecule-1 (ICAM-1), L selectin, and neutrophil endopeptidase 24.11 (NEP) in 38 patients with acute myocardial infarction (group 1) and 16 control subjects (group 2). In group 1, all the patients underwent blood sampling at initial presentation and 10 minutes later. Twenty of them had 3 mg of morphine administered intravenously immediately after the first sampling (group 1A) and the other 18 after a second sampling (group 1B). The serum levels of ICAM-1 and L-selectin were both significantly higher in groups 1A and 1B than in group 2. In group 1A, the ICAM-1 decreased significantly at second blood samplings (310 +/- 28 vs 368 +/- 30 ng/ml; p <0.001), whereas in group 1B there was no significant change in ICAM-1 (357 +/- 33 vs 359 +/- 26 ng/ml; p = NS). In group 1A, the L-selectin decreased significantly at second blood samplings (2.3 +/- 1.2 mg/L, p <0.001 vs baseline), whereas in group 1B there was no significant change in L-selectin (3.9 +/- 1.0 mg/L, p = NS vs baseline). There was no significant difference in baseline NEP activities between groups 1A and 1B (4.89 +/- 1.22 vs 5.14 +/- 1.57 nmol/mg protein; p = NS). However, the NEP activities at second blood samplings decreased significantly in group 1A (9.88 +/- 1.86 nmol/mg protein, p <0.001 vs baseline), whereas no significant changes were observed in group 1B (5.09 +/- 1.62 nmol/mg protein, p = NS vs baseline). In conclusion, morphine increased NEP activities and thus attenuated shedding of L-selectin and ICAM-1. PMID- 9416931 TI - Angioplasty guidewire velocity: a new simple method to calculate absolute coronary blood velocity and flow. AB - The Thrombolysis In Myocardial Infarction (TIMI) frame count is a relative index of coronary flow that measures time by counting the number of frames required for dye to travel from the ostium to a standardized coronary landmark in a cineangiogram filmed at a known speed (frames/s). We describe a new method to measure distance along arteries so that absolute velocity (length divided by time) and absolute flow (area x velocity) may be calculated in patients undergoing percutaneous transluminal coronary angiography (PTCA). After PTCA, the guidewire tip is placed at the coronary landmark and a Kelly clamp is placed on the guidewire where it exits the Y-adapter. The guidewire tip is then withdrawn to the catheter tip and a second Kelly clamp is placed on the wire where it exits the Y-adapter. The distance between the 2 Kelly clamps outside the body is the distance between the catheter tip and the anatomic landmark inside the body. Velocity (cm/s) may be calculated as this distance (cm) divided by TIMI frame count (frames) x film frame speed (frames/s). Flow (ml/s) may be calculated by multiplying this velocity (cm/s) and the mean cross-sectional lumen area (cm2) along the length of the artery to the TIMI landmark. In 30 patients, velocity increased from 13.9 +/- 8.5 cm/s before to 22.8 +/- 9.3 cm/s after PTCA (p <0.001). Despite TIMI grade 3 flow both before and after PTCA in 18 patients, velocity actually increased 38%, from 17.0 +/- 5.4 to 23.5 +/- 9.0 cm/s (p = 0.01). For all 30 patients, flow doubled from 0.6 +/- 0.4 ml/s before to 1.2 +/- 0.6 ml/s after PTCA (p <0.001). In the 18 patients with TIMI grade 3 flow both before and after PTCA, flow increased 86%, from 0.7 +/- 0.3 to 1.3 +/- 0.6 ml/s (p = 0.001). Distance along coronary arteries (length) can be simply measured using a PTCA guidewire. This length may be combined with the TIMI frame count to calculate measures of absolute velocity and flow that are sensitive to changes in perfusion. TIMI grade 3 flow is composed of a range of velocities and flows. PMID- 9416932 TI - Successful directional atherectomy of de novo coronary lesions assessed with three-dimensional intravascular ultrasound and angiographic follow-up. AB - Recent histopathologic and intravascular ultrasound (IVUS) data indicate that inadequate compensatory enlargement of atherosclerotic lesions contributes to the development of significant arterial stenoses. Such lesions may contain less plaque, which may have implications for atheroablative interventions. In this study, we compared lesions with (group A, n = 16) and without inadequate compensatory enlargement (group B, n = 30) as determined by IVUS. The acute results and the follow-up lumen dimensions of angiographically successful directional coronary atherectomy procedures were compared. Inadequate compensatory enlargement was considered present when the preintervention arterial cross-sectional area at the target lesion site was smaller than that at the (distal) reference site. Three-dimensional IVUS analysis and quantitative angiography were performed in 46 patients before and after intervention. IVUS measurements included the arterial, lumen, and plaque (arterial minus lumen) cross-sectional areas at the target lesion site (i.e., smallest lumen site) and the (distal) reference site. Angiographic follow-up was performed in 42 patients. Preintervention and postintervention angiographic measurements and IVUS lumen cross-sectional area measurements were similar in both groups. However, at follow up, the angiographic minimum lumen and reference diameters were significantly smaller in group A compared with group B (1.71 +/- 0.47 mm vs 2.14 +/- 0.73 mm, p <0.03, and 2.97 +/- 0.29 mm vs 3.39 +/- 0.76 mm, p <0.02; group A vs B). The data of this observational study suggest that lesions with inadequate compensatory enlargement, as determined by IVUS before intervention, may have less favorable long-term lumen dimensions after directional coronary atherectomy procedures. PMID- 9416933 TI - Effect of extended-release isosorbide mononitrate one hour after dosing in patients with stable angina pectoris. IMDUR Study Group. AB - The effect of extended-release isosorbide mononitrate (ER-ISMN) on exercise tolerance 1 hour after dosing was compared with that of placebo in a multicenter, randomized, double-blind study of 151 patients with stable effort-induced angina. During a 9- to 24-day placebo run-in, patients underwent Bruce protocol baseline exercise tolerance tests, after which they received ER-ISMN or placebo for 5 days. ER-ISMN patients took 60 mg each morning for the first 4 days and 120 mg on the morning of the fifth day. One hour after dosing, ER-ISMN patients had a significantly greater increase in total exercise time (days 1 to 4: 5 +/- 53 seconds; day 5: 53 +/- 58 seconds) than the placebo-treated patients (days 1 to 4: 14 +/- 37 seconds; day 5: 21 +/- 48) (p <0.001). The times to development of angina and 1-mm ST-segment depression were significantly longer in the ER-ISMN group than in the placebo group. The difference between the groups in mean time to onset of angina was 34 seconds after the 60-mg dose (p = 0.004) and 49 seconds after the 120-mg dose (p <0.001). The mean time to development of a 1-mm ST segment depression was 51 and 61 seconds longer after the 60-mg and 120-mg ER ISMN doses, respectively, than after placebo (p <0.001). Treatment-related adverse events were reported in 37% (28 of 75) and 7% (5 of 76) of patients in the ER-ISMN and placebo groups, respectively. As expected, headache was more frequent in the ER-ISMN group than in the placebo group (28% and 1%, respectively). The effects of ER-ISMN (60 mg and 120 mg) are clinically evident 1 hour after dosing, resulting in better exercise tolerance in patients with angina pectoris. PMID- 9416934 TI - Cost effectiveness of inpatient initiation of antiarrhythmic therapy for supraventricular tachycardias. AB - This study assessed the cost effectiveness of inpatient antiarrhythmic therapy initiation for supraventricular tachycardias using a metaanalysis of proarrhythmic risk and a decision analysis that compared inpatient to outpatient therapy initiation. A MEDLINE search of trials of antiarrhythmic therapy for supraventricular tachycardias was performed, and episodes of cardiac arrest, sudden or unexplained death, syncope, and sustained or unstable ventricular arrhythmias were recorded. A weighted average event rate, by sample size, was calculated and applied to a clinical decision model of therapy initiation in which patients were either hospitalized for 72 hours or treated as outpatients. Fifty-seven drug trials involving 2,822 patients met study criteria. Based on a 72-hour weighted average event rate of 0.63% (95% confidence interval, 0.2% to 1.2%), inpatient therapy initiation cost $19,231 per year of life saved for a 60 year-old patient with a normal life expectancy. Hospitalization remained cost effective when event rates and life expectancies were varied to model hypothetical clinical scenarios. For example, cost-effectiveness ratios for a 40 year-old without structural heart disease and a 60-year-old with structural heart disease were $37,510 and $33,310, respectively, per year of life saved. Thus, a 72-hour hospitalization for antiarrhythmic therapy initiation is cost effective for most patients with supraventricular tachycardias. PMID- 9416935 TI - Ethylene oxide on electrophysiology catheters following resterilization: implications for catheter reuse. AB - Reuse of electrophysiology catheters is an important cost-saving option for many laboratories. However, to be reused safely, catheters must undergo resterilization with ethylene oxide (EtO). Residual EtO levels on resterilized catheters may be high and could pose a risk to patients. Resterilized diagnostic electrophysiology catheters were tested for residual EtO using headspace gas chromatography after both a standard resterilization with an aeration process and after a resterilization process that incorporated a detoxification period. The Food and Drug Administration's maximum permissible level of EtO for implantable products, 25 parts per million (ppm), was used as the cutoff for acceptable catheter residuals. At day 2 after standard resterilization, the residual level of EtO on catheters was high at 41 +/- 6 ppm. However, these levels decreased with shelf time, decreasing to 26 +/- 3 ppm by day 7 and to 14 +/- 2 ppm by day 14 after sterilization, at which time all catheters were <25 ppm (p <0.001). Detoxification periods of 6, 12, and 15 hours were tested and 15 hours was found to be optimal. After 15 hours of detoxification, residual EtO was 19 +/- 1 ppm by day 2 and all catheters were <25 ppm. In summary, electrophysiology catheters that have undergone resterilization have residual EtO levels that are twice the Food and Drug Administration's limit for implantable products. Residual EtO levels may be substantially reduced either by allowing a 14-day waiting period after resterilization or by incorporating a detoxification period immediately after EtO exposure. PMID- 9416936 TI - Predischarge arrhythmia induction testing of implantable defibrillators may be unnecessary in selected cases. AB - Complete postoperative evaluation of implantable cardioverter-defibrillators (ICDs) before discharge, including arrhythmia induction, has been the standard since their introduction. Whereas the original ICDs provided little telemetered information and used separate pace-sense and defibrillation leads, modern, third generation devices provide pace-sense function information in addition to other data and are used in conjunction with integrated transvenous endocardial leads that combine pace-sense and defibrillation function. Changes in lead position, which can potentially result in either an inability to detect fibrillation or to terminate it, should be mirrored by changes in resting pace-sense function. Thus, for newer ICDs implanted with integrated endocardial lead systems, it is possible that in at least some cases predischarge arrhythmia inductions can be avoided. Two hundred patients receiving third-generation ICDs in conjunction with integrated transvenous leads were evaluated before discharge. Defibrillation detection or termination problems were seen in 8. Declines in resting R-wave amplitude and pacing impedance were significantly associated with such complications (-7 +/- 5 vs -0.3 +/- 2.3 mV [p <0.0001] and -158 +/- 138 vs -93 +/ 76 omega [p <0.05], for those with vs without complications, respectively), as were gross right ventricular lead migrations on chest x-ray. No patient with a defibrillation complication had an R-wave change of <3 mV. However, 13% of patients without complications had R-wave changes of >3 mV. It is concluded that a pace-sense evaluation of ICDs may be a satisfactory screen to determine those who need to go on to complete testing with arrhythmia induction in selected cases. PMID- 9416937 TI - Relation of endothelium, thrombogenesis, and hemorheology in systemic hypertension to ethnicity and left ventricular hypertrophy. AB - Although the arterial tree is exposed to increased pressure in hypertensive patients, paradoxically, the complications of hypertension (heart attacks, stroke) are mainly thrombotic rather than hemorrhagic. Patients with left ventricular (LV) hypertrophy are at high risk of the complications of hypertension. We performed a cross-sectional study of 178 patients attending a hypertension clinic in a city center teaching hospital, and measured plasma levels of the soluble adhesion molecule P-selectin (associated with platelet activity/function and atherosclerosis), the von Willebrand factor (vWf; a marker of endothelial dysfunction), fibrin D-dimer (an index of thrombogenesis), plasminogen activator inhibitor (PAI, an index of fibrinolysis), lipoprotein(a) (Lp(a), associated with thrombogenesis and atherogenesis) and hemorheological indexes (fibrinogen, hematocrit, plasma viscosity, hemoglobin) in patients with essential hypertension, in whom the LV mass and LV mass index were determined using echocardiography. The 178 patients (86 men, mean age 54 +/- 15 years) were compared with 47 normotensive healthy controls (aged 56 +/- 20 years). Hypertensive patients had higher P-selectin, PAI, vWf, fibrin D-dimer, Lp(a), plasma fibrinogen, and plasma viscosity when compared with controls. Black hypertensive patients had higher Lp(a) levels and LV septal and posterior wall thickness on echocardiography, but lower plasma PAI levels. Patients with LV hypertrophy (defined as a LV mass index > 134 g/m2 in men or > 110 g/m2 in women) had higher plasma fibrinogen compared with those without LV hypertrophy. Systolic blood pressures were significantly correlated to age, plasma viscosity, plasma fibrinogen, and vWf. Diastolic blood pressures were significantly correlated with age and plasma fibrinogen. Fibrinogen levels were correlated with LV mass, LV mass index, left atrial size, plasma viscosity, and vWf. Fibrin D-dimer levels were significantly correlated with vWf and fibrinogen levels. Thus, hypertensive patients have high plasma fibrinogen levels, thrombogenesis, and impaired fibrinolysis (as indicated by high D-dimer and PAI levels, respectively), platelet activation (raised soluble P-selectin), and endothelial dysfunction (high vWF). The high plasma fibrinogen levels were related to blood pressures, LV mass index (and LV hypertrophy), and left atrial size. These abnormalities in hemorheologic factors and markers of thrombogenesis and endothelial function may act synergistically to increase the risk of thrombogenesis and atherosclerosis in hypertensive patients. PMID- 9416938 TI - Comparison of changes in respiratory function and exercise oxygen uptake with losartan versus enalapril in congestive heart failure secondary to ischemic or idiopathic dilated cardiomyopathy. AB - In congestive heart failure (CHF), some of the effects of angiotensin-converting enzyme (ACE) inhibitors, such as an increase in exercise oxygen uptake (VO2), are mediated through prostaglandins. Angiotensin (AT1) receptor blockers apparently do not share potentiation of this biosystem. We tested whether losartan improves exercise VO2 in CHF and if the effect is the same as for enalapril. Sixteen men with CHF and 8 volunteers, all nonsmokers and not taking ACE, AT1 receptor, or cyclooxygenase inhibitors, were randomized to receive placebo, enalapril (10 mg 2 times daily), losartan (50 mg/day), each of these 2 drugs plus aspirin (325 mg/day), aspirin, or the same preparations in a reverse order, each for 3 weeks, with a 3-week washout period between treatments. Pulmonary function and VO2 were assessed at the end of each treatment. In CHF, losartan and enalapril caused a similar improvement of VO2 and exercise tolerance, which was absent in controls and was counteracted by aspirin (prostaglandin inhibition) when obtained with enalapril and not with losartan. While on enalapril, we also detected an increase in the diffusing lung capacity for carbon monoxide, which correlated with changes in VO2 and was antagonized by aspirin, suggesting the possibility that a prostaglandin-mediated functional improvement of the alveolar capillary membrane contributes to the rise in VO2. Thus, losartan is as effective as enalapril for exercise VO2 and exercise tolerance, but the mechanism seems to be dissociated from a prostaglandin biosystem activation. Losartan may represent an advancement in CHF because its efficacy on VO2 is similar to that of enalapril, but is not antagonized by aspirin. PMID- 9416939 TI - Feasibility, accuracy, and incremental value of intraoperative three-dimensional transesophageal echocardiography in valve surgery. AB - In this prospective trial, intraoperative 2-dimensional (2-D) and 3-dimensional (3-D) transesophageal echocardiography (TEE) examinations were performed on 60 consecutive patients undergoing cardiac valve surgery. Both 2-D (including color flow and Doppler data) and 3-D images were reviewed by blinded observers, and major valvular morphologic findings recorded. In vivo morphologic findings were noted by the surgeon and all explanted valves underwent detailed pathologic examination. To test reproducibility, 6 patients also underwent 3-D TEE 1 day before surgery. A total of 132 of 145 attempted acquisitions (91%) were completed with a mean acquisition time of 2.8 +/- 0.2 minutes. Acquisition time was significantly shorter in patients with regular rhythms. Reconstructions were completed in 121 of 132 scans (92%) and there was at least 1 good reconstruction in 56 of 60 patients (93%). Mean reconstruction time was 8.6 +/- 0.7 minutes. Mean effective 3-D time, which was the time taken to complete an acquisition and a clinically interpretable reconstruction, was 12.2 +/- 0.8 minutes. Intraoperative 3-D echocardiography was clinically feasible in 52 patients (87%). Three-D echocardiography detected most of the major valvular morphologic abnormalities, particularly leaflet perforations, fenestrations, and masses, confirmed on pathologic examination. Three-D echocardiography predicted all salient pathologic findings in 47 patients (84%) with good quality images. In addition, in 15 patients (25%), 3-D echocardiography provided new additional information not provided by 2-D echocardiography, and in 1 case, 3-D echocardiographic findings resulted in a surgeon's decision to perform valve repair rather than replacement. In several instances, 3-D echocardiography provided complementary morphologic information that explained the mechanism of abnormalities seen on 2-D and color flow imaging. In the reproducibility subset, preoperative and intraoperative 3-D imaging detected a similar number of findings when compared with pathology. Thus, in routine clinical intraoperative settings, 3-dimensional TEE is feasible, accurately predicts valve morphology, and provides additional and complementary valvular morphologic information compared with conventional 2-D TEE, and is probably reproducible. PMID- 9416940 TI - Comparison of mortality rates and progression of left ventricular dysfunction in patients with idiopathic dilated cardiomyopathy and dilated versus nondilated right ventricular cavities. AB - This study assesses the influence of right ventricular (RV) dilation on the progression of left ventricular (LV) dysfunction and survival in patients with idiopathic dilated cardiomyopathy (IDC). Using transthoracic echocardiography, we studied 100 patients with IDC aged 20 to 80 years (mean 55 +/- 14); 67% were men. In the apical 4-chamber view, diastolic LV and RV chamber area measurements classified patients into 2 groups: group RV enlargement+ (RV area/LV area > 0.5) included 54 patients; group RV enlargement- (no RV enlargement) had RV area/LV area < or = 0.5. Echocardiographic studies were repeated in all patients after a mean of 33 +/- 16 months. At the time of the initial study, the 2 groups did not differ in age, gender, incidence of atrial fibrillation and diabetes, left ventricular mass, and LV ejection fraction, but the RV enlargement+ group had more severe tricuspid regurgitation and less LV enlargement. After 47 +/- 22 months (range 12 to 96), patients in group RV enlargement+ had lower LV ejection fraction (29% vs 34%, p = 0.006) than patients with initial RV enlargement-. At clinical follow-up, mortality was higher (43%) in patients with initial RV enlargement+ than the RV enlargement- patients (15%), p = 0.002. For survivors, the mitral deceleration time averaged 157 +/- 36 ms; for nonsurvivors or patients who required transplant, the mitral deceleration time averaged 97 +/- 12 ms (p < 0.0001). With use of a multivariate Cox model adjusting for LV ejection fraction, LV size, and age, the relative risk ratio of mortality from initial RV enlargement+ was 4.4 (95% confidence limits 1.7 to 11.1) (p = 0.002). Thus, patients with significant RV dilation had nearly triple the mortality over 4 years and more rapidly deteriorating LV function than patients with less initial RV dilation. In IDC, RV enlargement is a strong marker for adverse prognosis that may represent a different morphologic subset. PMID- 9416941 TI - Why does thrombolysis fail? Breaking through the reperfusion ceiling. AB - With best current regimens, thrombolysis may initially fail in 15% to 40% of cases because of hemodynamic or mechanical factors, inadequate fibrinolysis, or failure to lyse platelet-rich thrombi. Pathophysiologic considerations and early experimental and clinical trials suggest that improved conjunctive antiplatelet therapy (as with glycoprotein IIb/IIIa inhibitors) and, possibly, more effective antithrombins (low molecular weight heparins, hirudin) may allow this thrombolysis "ceiling" to be broken and the goal achieved of reperfusion rates closer to those of primary angioplasty. PMID- 9416942 TI - Short-term (4 hours) observation after elective coronary angioplasty. AB - In a prospective evaluation of the safety of short-term observation after elective percutaneous transluminal coronary angioplasty (PTCA) in 1,900 consecutive patients, 1 of 1,680 patients triaged to discharge after 4 hours of observation reached the primary end point of acute recurrent ischemia, 7 patients underwent repeat PTCA during 4 hours of observation, and 66 of 187 patients selected for prolonged observation had a complicated course. It is concluded that short-term observation after elective coronary angioplasty is safe, with a negligible risk of vessel closure after this period; triage for prolonged observation can be based appropriately on the immediate procedural result. PMID- 9416944 TI - Mechanisms of limited maximum coronary flow in severe single-vessel coronary artery disease in humans due to vertical steal. AB - We evaluated the usefulness of a decrease in the average peak velocity from 4 to 10 minutes after infusion of dipyridamole for detecting myocardial ischemia in 50 patients, including patients with a prior myocardial infarction. The decrease in the average peak velocity from 4 to 10 minutes associated with vertical steal and combined with a coronary flow reserve of < 1.6 had a high predictive value for myocardial ischemia in patients with or without prior myocardial infarction. PMID- 9416943 TI - Effects of quinapril on coronary blood flow in coronary artery disease patients with endothelial dysfunction. TREND Investigators. Trial on Reversing Endothelial Dysfunction. AB - This pilot study evaluates the effects of quinapril, an angiotensin-converting enzyme inhibitor with high tissue-binding affinity, on microvascular endothelial function in patients with mild (<40% narrowing) coronary artery disease and epicardial endothelial dysfunction. Patients randomized to quinapril had a trend suggesting an increase in endothelium-dependent coronary blood flow response. PMID- 9416945 TI - Results of Micro stent implantations in coronary lesions of various complexity. AB - Micro stents appear to be especially suitable for the safe treatment of complex coronary lesions and adverse vessel morphology. Stenting of lesions with type C morphology is associated with a higher restenosis rate than stenting of less complex coronary obstructions. PMID- 9416946 TI - Assessment of left internal mammary artery grafts using dipyridamole Doppler echocardiography. AB - Color Doppler echocardiography of the left mammary artery was combined with dipyridamole testing in order to assess the presence of significant (>70%) graft stenosis in 87 patients with a mammary artery graft to the left anterior descending coronary artery presenting with chest pain. Occluded grafts are detected by absent diastolic flow velocities at baseline, whereas the response of the diastolic flow velocity to dipyridamole distinguishes patients with critical versus noncritical stenosis of a patent graft. PMID- 9416947 TI - A prospective hemodynamic evaluation of patients with chronic atrial fibrillation undergoing radiofrequency catheter ablation of the atrioventricular junction. AB - A prospective invasive hemodynamic evaluation in 11 unselected patients with medically refractory chronic atrial fibrillation undergoing radiofrequency catheter ablation of the atrioventricular junction was performed. The resultant rate regulation and control caused a hemodynamic and symptomatic improvement despite persistent fibrillation at the atrial level. PMID- 9416948 TI - Long-term follow-up after radiofrequency ablation of paroxysmal supraventricular tachycardia in patients with tachycardia-induced atrial fibrillation. AB - Four of 12 patients (33%) with paroxysmal supraventricular tachycardia (PSVT) and tachycardia-induced atrial fibrillation (AF) had recurrences of paroxysmal AF after successful catheter ablation of the PSVT. This study demonstrates that AF often remains a problem after radiofrequency catheter ablation of PSVT in patients with tachycardia-induced AF, and it may not be possible to predict in which patients this will be the case. PMID- 9416949 TI - Predictors of crossover to a left ventricular approach for atrioventricular junction ablation. AB - In a retrospective analysis of patients referred for atrioventricular node radiofrequency ablation, male gender and a history of hypertension were found to be predictors of crossover to a left ventricular approach for success. This subgroup of patients may benefit from early crossover if initial attempts at right-sided ablation fail. PMID- 9416950 TI - Effects and tolerability of irbesartan versus enalapril in patients with severe hypertension. Irbesartan Multicenter Investigators. AB - In this double-blind, randomized study, an antihypertensive regimen based on irbesartan, an angiotensin II receptor antagonist, reduced systolic and diastolic blood pressure by 40/30 mm Hg at week 12 in patients with severe hypertension; this reduction was at least equivalent to that of a regimen using enalapril up to 40 mg. The irbesartan-based regimen had a better tolerability profile with fewer adverse events (55% vs 64%) and significantly less cough (2.5% vs 13.1%, p = 0.007). PMID- 9416952 TI - Effects of atrial fibrillation on left ventricular function and geometry in mitral stenosis. AB - This study shows that patients with mitral stenosis have depressed left ventricular ejection performance and spherical remodeling of the left ventricular cavity, which is more marked in those with atrial fibrillation. These changes have important clinical implications regarding treatment strategy in patients with mitral stenosis and chronic atrial fibrillation. PMID- 9416951 TI - Comparison of estimates of right atrial pressure by physical examination and echocardiography in patients with congestive heart failure and reasons for discrepancies. AB - Clinical estimates of right atrial pressure from the jugular venous pulse were accurate when right atrial pressure was normal, but systematically underestimated elevated right atrial pressures. Because the increased distance from the mid right atrium to the sternal angle is not accounted for, apparently normal right atrial pressure estimates by this technique do not reliably exclude elevated right atrial pressure in patients with congestive heart failure. PMID- 9416953 TI - Comparison of coronary vasodilator reserve in elite rowing athletes versus hypertrophic cardiomyopathy. AB - Compared to normal volunteers, coronary vasodilation reserve is reduced in patients with hypertrophic cardiomyopathy but not in rowing athletes with left ventricular hypertrophy. Positron emission tomography can provide complementary information to distinguish between the athlete's heart and hypertrophic cardiomyopathy. PMID- 9416954 TI - Clinical implications and possible association of malposition of the branch pulmonary arteries with DiGeorge syndrome and microdeletion of chromosomal region 22q11. AB - We describe a series of 10 patients with malposition of the branch pulmonary arteries (4 patients with crossing [crossed pulmonary arteries] and 6 patients without crossing), 2 of whom had a short main pulmonary artery segment that resulted in iatrogenic right pulmonary artery stenosis after pulmonary artery band placement. DiGeorge syndrome was seen in 5 patients and 4 had microscopic deletion of chromosomal region 22q11. PMID- 9416955 TI - Inhibition of human low-density lipoprotein oxidation by flavonoids in red wine and grape juice. AB - In the presence of red wine or grape juice, low-density lipoprotein was significantly resistant to oxidation; the biological activity of flavonoids, but not ethanol or nonflavonoid phenolic compounds, appeared to contribute to the antioxidant properties of red wine and grape juice. A significant antioxidant activity was also confirmed in low-density lipoprotein from humans after ingesting red wine but not grape juice, suggesting that flavonoids in red wine can be absorbed from the intestine more efficiently than those in grape juice. PMID- 9416956 TI - Improved atrial function in bicaval versus standard orthotopic techniques in cardiac transplantation. AB - Atrial geometry is preserved in the bicaval technique of cardiac transplantation. Using Doppler echocardiography, we investigated the impact of this technique on preservation of atrial function and found that echocardiographic indexes of atrial function are improved in bicaval cardiac transplants versus the standard orthotopic transplants. PMID- 9416957 TI - Percutaneous balloon pericardiotomy for patients with recurrent pericardial effusion: using a novel double-balloon technique with one long and one short balloon. AB - Percutaneous balloon pericardiotomy is effective and less invasive for the treatment of recurrent pericardial effusion. This study suggests that the double balloon method with 1 longer and 1 shorter balloon is the procedure of choice for percutaneous balloon pericardiotomy. PMID- 9416958 TI - Measurement of coronary flow reserve in children by transthoracic Doppler echocardiography. AB - Noninvasive measurement of coronary flow reserve was performed by transthoracic color Doppler echocardiography in 28 children with Kawasaki disease. PMID- 9416959 TI - Effects of exercise-induced oxidative stress on nitric oxide release and antioxidant activity. AB - This study shows that acute exercise in healthy subjects is a modest oxidative stress, which may be related to an increase in antioxidant activity and down regulation of nitric oxide formation. PMID- 9416960 TI - Enalapril and QTC dispersion in congestive heart failure. PMID- 9416962 TI - Anulus, instead of annulus. PMID- 9416961 TI - Werner Forssmann and the Nazis. PMID- 9416963 TI - Late coronary abnormalities after arterial switch operation for transposition of the great arteries. PMID- 9416964 TI - Atorvastatin versus simvastatin. PMID- 9416965 TI - Anti-heart autoantibodies are more frequently present in Chagas disease patients with dilated cardiomyopathy. PMID- 9416966 TI - Clinical implication of the hyperventilation test in the diagnosis of coronary artery spasm. PMID- 9416967 TI - New horizons in the treatment of tuberculosis. AB - The development of new chemotherapy for the treatment of tuberculosis has three major objectives: first, the development of faster-acting drugs to shorten the duration of treatment; second, the development of novel antimicrobials to counter the emergence of bacteria resistant to current therapies; and, third, the development of chemotherapeutics that specifically target dormant bacilli to treat the one-third of the world's population latently infected with tubercle bacilli. Strategies based upon optimizing the inhibition of known targets require an extensive knowledge of the detailed mechanism of action of current antimycobacterial agents. For many agents such as isoniazid, ethambutol, rifampin, and pyrazinamide such knowledge is now available. Strategies based upon the identification of novel targets will necessitate the identification of biochemical pathways specific to mycobacteria and related organisms. Many unique metabolic processes occur during the biosynthesis of mycobacterial cell wall components, and some attractive new targets have emerged. The development of targets specific to latency will require a detailed picture of the metabolism and biochemical pathways occurring in dormant bacilli. Recent evidence suggests that anaerobic metabolic pathways may operate in dormant bacilli, and the enzymes involved in such pathways may also provide significant new targets for intervention. The combination of the mycobacterial genome sequence that is anticipated to become available this year with an improved understanding of the unique metabolic processes that define mycobacteria as a genus offers the greatest hope for the elimination of one of mankind's oldest enemies. PMID- 9416968 TI - EB 1089, a novel vitamin D analog with strong antiproliferative and differentiation-inducing effects on target cells. AB - The physiologically active form of vitamin D, 1alpha,25-dihydroxyvitamin D3, plays an important role not only in the establishment and maintenance of calcium metabolism, but also in regulating cell growth and differentiation. As the clinical usefulness of 1alpha,25-dihydroxyvitamin D3 is limited by its tendency to cause hypercalcemia, new analogs with a better therapeutic profile have been synthesized. One of these new synthetic vitamin D analogs is EB 1089, which is characterized by an altered side chain structure featuring 26,27-dimethyl groups and two double bonds. This analog has been shown to be more potent than 1,25 dihydroxyvitamin D3 in inhibiting proliferation, stimulating differentiation, and inducing apoptosis in a number of different cell types, including cancer cells. Despite being more potent than 1alpha,25-dihydroxyvitamin D3 with respect to its cell regulatory effects, EB 1089 displays weaker calcemic side-effects. These characteristics make EB 1089 a potentially useful compound for the treatment of a diversity of clinical disorders, including cancer and metabolic bone diseases. A promising phase I study with EB 1089 in patients with advanced breast and colon cancer has already been carried out, and more clinical trials evaluating the clinical effectiveness of EB 1089 in other types of cancer are in progress. PMID- 9416969 TI - Effects on DNA integrity and apoptosis induction by a novel antitumor sesquiterpene drug, 6-hydroxymethylacylfulvene (HMAF, MGI 114). AB - 6-Hydroxymethylacylfulvene (HMAF, MGI 114) is a new alkylating antitumor sesquiterpenoid with promising and often curative antitumor activity in vivo. This study examined the ability of the drug to damage cellular DNA, induce apoptosis, and affect the cell cycle of CEM human leukemia cells. No bifunctional lesions, interstrand DNA cross-links or DNA-protein cross-links were seen (by alkaline sedimentation and K+/SDS precipitation, respectively) when using up to 50 microM HMAF. The drug possibly formed some monoadducts, as DNA from drug treated cells impeded primer extension by Taq polymerase, although only partial inhibition was seen even at 200 microM HMAF. HMAF also induced secondary lesions in cellular DNA, single-strand breaks that were detectable (by nucleoid sedimentation and alkaline sucrose gradient analysis) after a 4-hr treatment at HMAF levels as low as 2 microM, comparable to the growth inhibition IC50 value (1.7 microM). A post-treatment incubation of cells in drug-free medium generated substantial amounts of DNA double-stranded fragments of several kbp, suggesting apoptotic fragmentation (>30% of total DNA following treatment with 20 microM HMAF and a 17-hr post-treatment incubation). Chromatin condensation (by ultrastructural analysis) and induction of sub-G1 particles and apoptotic strand breakage (by multiparametric flow cytometry) confirmed induction of apoptosis by HMAF. HMAF preferentially inhibited DNA synthesis (IC50 approximately 2 microM), which is consistent with an S phase block, observed by cell cycle analysis. The pattern of apoptotic DNA fragmentation, inhibition of DNA synthesis, and blockage in the S phase suggests that these events play a role in the antiproliferative activity of HMAF. PMID- 9416970 TI - Human liver cytochrome P450 enzymes involved in the 7-hydroxylation of R- and S warfarin enantiomers. AB - Human liver microsomes had about 8-fold higher 7-hydroxylation activities for S warfarin than for R-warfarin. Activities of racemic warfarin 7-hydroxylation by liver microsomes of 35 human samples correlated more closely with those of S warfarin 7-hydroxylation (r = 0.95) than with those of R-warfarin 7-hydroxylation (r = 0.69). The correlation coefficient between R-warfarin 7-hydroxylation and 7 ethoxyresorufin O-deethylation activities was 0.73 in these human samples, suggesting that R- and S-warfarin enantiomers are catalyzed by different forms of human cytochrome P450 (P450 or CYP) enzymes. Anti-CYP2C9 antibodies inhibited completely the 7-hydroxylation of S-warfarin, but not R-warfarin, catalyzed by human liver microsomes, while anti-CYP1A2 inhibited R-warfarin 7-hydroxylation by about 70%. Interestingly, the racemic warfarin 7-hydroxylation activities (turnover numbers of 1.6 +/- 1.0 pmol/min/mg protein in 35 human samples) were found to be low compared with the S-warfarin 7-hydroxylation activities (4.1 +/- 2.5 pmol/min/mg protein), indicating that R-warfarin may have affected the CYP2C9 dependent S-warfarin 7-hydroxylation activities when racemic warfarin was used as a substrate. Several P450 inhibitors, as well as R-warfarin, were examined for their abilities to inhibit S-warfarin 7-hydroxylation; we found that R-warfarin was a non-competitive inhibitor with a Ki value of about 150 microM, whereas both tolbutamide and sulfaphenazole were competitive inhibitors with Ki values of about 100 and 0.5 microM, respectively, for S-warfarin 7-hydroxylation activities. These results suggest that R- and S-warfarin enantiomers are catalyzed principally by CYP1A2 and CYP2C9, respectively, in human liver microsomes, and that the pharmacokinetic properties of S-warfarin may be altered by R-warfarin in vivo when racemic warfarin is administered clinically to humans. PMID- 9416971 TI - Antifibrotic effect of decorin in a bleomycin hamster model of lung fibrosis. AB - We reported previously that treatment with antibody to transforming growth factor beta (TGF-beta) caused a marked attenuation of bleomycin (BL)-induced lung fibrosis (LF) in mice. Decorin (DC), a proteoglycan, binds TGF-beta and thereby down-regulates all of its biological activities. In the present study, we evaluated the antifibrotic potential of DC in a three-dose BL-hamster model of lung fibrosis. Hamsters were placed in the following groups: (1) saline (SA) + phosphate-buffered saline (PBS) (SA + PBS); (2) SA + DC; (3) BL + PBS; and (4) BL + DC. Under pentobarbital anesthesia, SA (4 mL/kg) or BL was instilled intratracheally in three consecutive doses (2.5, 2.0, 1.5 units/kg/4 mL) at weekly intervals. DC (1 mg/mL) or PBS was instilled intratracheally in 0.4 mL/hamster on days 3 and 5 following instillation of each dose of SA or BL. In week 4, hamsters received three doses of either DC or PBS every other day. The hamsters were killed at 30 days following the first instillation, and their lungs were appropriately processed. Lung hydroxyproline levels in SA + PBS, SA + DC, BL + PBS, and BL + DC groups were 965, 829, 1854, and 1387 microg/lung, respectively. Prolyl hydroxylase activities were 103, 289, and 193% of SA + PBS control in SA + DC, BL + PBS, and BL + DC groups, respectively. The myeloperoxidase activities in the corresponding groups were 222, 890, and 274% of control (0.525 units/lung). Intratracheal instillation of BL caused significant increases in these biochemical markers, and instillation of DC diminished these increases in the BL + DC group. DC treatment also caused a significant reduction in the infiltration of neutrophils in the bronchoalveolar lavage fluid (BALF) of hamsters in the BL + DC group. However, DC treatment had little effect on BL induced increases in lung superoxide dismutase activity and lipid peroxidation and leakage of plasma proteins in the BALF of the BL + DC group. Hamsters in the BL + PBS group showed severe multifocal fibrosis and accumulation of mononuclear inflammatory cells and granulocytes. In contrast, hamsters in the BL + DC group showed mild multifocal septal thickening with aggregations of mononuclear inflammatory cells. Hamsters in both control groups (SA + PBS and SA + DC) showed normal lung structure. Frozen lung sections following immunohistochemical staining revealed an intense staining for EDA-fibronectin and collagen type I in the BL + PBS group as compared with all other groups. It was concluded that DC potentially offers a novel pharmacological intervention that may be useful in treating pulmonary fibrosis. PMID- 9416972 TI - Bioreductive metabolism of the novel fluorinated 2-nitroimidazole hypoxia probe N (2-hydroxy-3,3,3-trifluoropropyl)-2-(2-nitroimidazolyl) acetamide (SR-4554). AB - The aim of this work was to study the metabolic characteristics of the novel fluorinated 2-nitroimidazole hypoxia probe N-(2-hydroxy-3,3,3-trifluoropropyl)-2 (2-nitroimidazolyl) acetamide (SR-4554). HPLC and 19F NMR methods were employed to evaluate the rate of reductive metabolism of SR-4554 and the nature of the resulting metabolites, respectively. SR-4554 was enzymatically reduced by mouse liver microsomes (1.1 +/- 0.1 nmol of SR-4554 reduced/min/mg protein), purified rat and human NADPH:cytochrome P450 reductase (17.8 +/- 0.4 and 5.0 +/- 0.5 nmol of SR-4554 reduced/min/mg protein, respectively), and SCCVII tumour homogenates (2.3 +/- 0.3 nmol of SR-4554 reduced/min/g tumour) under nitrogen. NADPH:cytochrome P450 reductase was a major microsomal enzyme involved in the bioreduction of SR-4554 by liver microsomes. In a panel of murine and human tumour xenografts, cytochrome P450 reductase activities were found to be low and only varied by 3-fold between different tumour types, suggesting that enzyme activities within the tumours are unlikely to influence markedly in vivo reductive metabolism. Reduction of SR-4554 by mouse liver microsomes showed a characteristic oxygen dependence with a half-maximal inhibition of 0.48 +/- 0.06%. Thus, the reductive metabolism of SR-4554 can be employed to detect the low oxygen tensions that occur within both murine and human tumours. Soluble, low molecular weight reductive metabolites of SR-4554 were identified by 19F NMR. These metabolite peaks appeared (up to 0.12 ppm) downfield of the parent drug peak. In conclusion, SR-4554 undergoes an oxygen-dependent metabolism that involves NADPH:cytochrome P450 reductase. 19F NMR is capable of identifying reduced metabolites that are undetectable by HPLC. PMID- 9416973 TI - Effects of proton pump inhibitors on thyroid hormone metabolism in rats: a comparison of UDP-glucuronyltransferase induction. AB - The effects of proton pump inhibitors on thyroid hormone metabolism in rats were examined. Pantoprazole, omeprazole, and lansoprazole were administered repeatedly to female SD rats at doses of 5, 50, and 300 mg/kg/day for 1 week, and changes in UDP-glucuronyltransferase activities were examined. Increases in o-aminophenol UDP-glucuronyltransferase activity, which was measured as that responsible for the glucuronidation of thyroxine, were evident following 7-day high-dose administration of all the proton pump inhibitors tested. Of the three proton pump inhibitors investigated, o-aminophenol UDP-glucuronyltransferase activity was greatest following the high-dose administration of omeprazole. Androsterone UDP glucuronyltransferase activity in rats treated with the proton pump inhibitors increased significantly, but these increases were smaller than those of o aminophenol UDP-glucuronyltransferase. Pantoprazole and omeprazole treatment did not affect plasma T4 or T3 significantly, whereas lansoprazole treatment produced marked reductions in plasma T4 but did not affect plasma T3 significantly. After administration of 125I-labeled thyroid hormone to rats treated with the proton pump inhibitors, biliary excretion of radioactivity increased significantly in omeprazole- and lansoprazole-treated rats; these increases were attributed to induction of liver thyroxine UDP-glucuronyltransferase activities. The order of biliary excretion of radioactivity, as well as the o-aminophenol UDP glucuronyltransferase activity, in the treated animals was: omeprazole > lansoprazole > pantoprazole. Therefore, repeated administration of the proton pump inhibitors increased thyroxine-metabolizing activity via induction of UDP glucuronyltransferase, and this induction by pantoprazole was less pronounced than that by omeprazole or lansoprazole. PMID- 9416974 TI - Transcription and activity of 5-fluorouracil converting enzymes in fluoropyrimidine resistance in colon cancer in vitro. AB - Cellular resistance to 5-fluorouracil (5-FU) is not completely understood. Since 5-FU shares the pyrimidine pathway with the physiological pyrimidines, we investigated the relationship between fluoropyrimidine metabolism, nucleic acid uptake and cytotoxicity of 5-FU in eight colon tumour cell lines including 5-FU resistant subclones. The cytotoxicity of 5-FU was increased up to 423-fold when the anabolites 5-fluorouridine (FUrd), 5-fluorodeoxyuridine (FdUrd), and 5 fluorodeoxyuridine monophosphate (FdUMP) were compared with the parent drug in vitro. The enzymes uridine phosphorylase and thymidine phosphorylase were predictive for the cytotoxicity of 5-FU in 5/7 cell lines. Inhibition of uridine phosphorylase and thymidine phosphorylase by antisense strategies effectively antagonised 5-FU, abolishing 84% and 79% of its toxicity. The importance of thymidine phosphorylase was supported by a highly restricted enzyme activity in 5 FU-resistant cells. In 5-FU naive cells, a stimulating effect of 5-FU on thymidylate synthase mRNA and ribonucleotide reductase mRNA expression was observed. In these cells, antisense oligonucleotides to ribonucleotide reductase significantly reduced cell growth. Downregulation of ribonucleotide reductase mRNA in 5-FU-resistant subclones suggests different mechanisms in primary and secondary resistance to 5-FU. Most of the intracellular 5-FU was selectively incorporated into RNA (range: 45-91%) and generally spared DNA (range: 0.2-11%). In synthesising our data, we conclude that drug resistance could be overwhelmed through bypassing limiting steps in the activation of 5-FU. In the majority of colonic tumours, the activity of uridine phosphorylase and thymidine phosphorylase may have prognostic relevance for the cytotoxicity of 5-FU in vitro. PMID- 9416975 TI - Differences in the stimulation of the phosphoinositide cycle by amine neurotransmitters in cultured rat forebrain neurones and astrocytes. AB - In this study, we compared the stimulation by carbachol (CCh), noradrenaline (NA), and histamine (HA) of phosphoinositide hydrolysis in rat forebrain neuronal and glial cultures. When Ca2+ was omitted from the stimulation buffer (low microM extracellular Ca2+), amine-induced [3H]inositol phosphate accumulation was reduced to a higher extent in astrocytes (70-80% for CCh and NA and 100% for HA) than in neurones (around 50-60% for all the amines). Furthermore, guanosine 5' [gamma-thio]trisphosphate (GTP[S]) stimulation of phosphoinositidase C (PIC) in membranes was 5-fold higher in neurones than in astrocytes. These results indicate differences in the mechanism of PIC stimulation in the two cell types. After 30 min stimulation in the presence of 10 mM Li+, a higher accumulation of [3H]inositol 4-monophosphate and [3H]inositol 1,4-bisphosphate than of [3H]inositol 1/3-monophosphate occurred for all agonists in neurones, whereas the opposite was observed in astrocytes. Moreover, in these cells stimulation for 5 min in the absence of Li+ produced a 2-3-fold accumulation of all metabolites of the 3-kinase pathway of inositol-1,4,5-trisphosphate metabolism but not of those of the 5-phosphatase pathway. Thus, regardless of the amine receptor stimulated, the 3-kinase route appeared to prevail in astrocytes and the 5-phosphatase pathway in neurones. The histamine response in neurones differed from that of the other agonists in that it rapidly declined. Taken together these results indicate that the heterogeneity in amine stimulation of the phosphoinositide cycle previously observed in brain slices could arise to a great extent from the cellular diversity of this preparation and be related to the differential contribution of the amine receptors located in neurones and astrocytes. PMID- 9416976 TI - Inhibition of human mitochondrial aldehyde dehydrogenase by the disulfiram metabolite S-methyl-N,N-diethylthiocarbamoyl sulfoxide: structural characterization of the enzyme adduct by HPLC-tandem mass spectrometry. AB - S-Methyl-N,N-diethylthiocarbamoyl sulfoxide (MeDTC-SO) is a known metabolite of the aversion therapy drug disulfiram (DSF). MeDTC-SO is also a potent inhibitor of human mitochondrial aldehyde dehydrogenase (hmALDH) with an IC50 of 1.5 microM. Inhibition of the enzyme by MeDTC-SO resulted in the addition of approximately 100 Da to the molecular mass of the intact protein, as determined by on-line HPLC-electrospray ionization MS (LC-MS). Dialysis of the inhibited protein did not reverse the inhibition, and the molecular mass of 54,533 Da (+/- 0.01%) remained unchanged, indicating that a covalent modification of the protein had occurred. Proteolytic digestion of hmALDH under basic conditions using trypsin at pH 7.8 revealed that the adduct was base labile. However, treating the adducted protein with endopeptidase-Glu-C at pH 3.7 produced a peptide adduct at MH+ = 4924, tentatively attributable to a carbamoylated peptide. This peptide contains three adjacent cysteines, one of which has been implicated as a key amino acid in the highly conserved active site region of ALDH. A pepsin digestion of hmALDH carried out at pH 3.7 and subsequent LC-MS analysis revealed an ion at MH2(2+) = 501.5, corresponding to the carbamoylated peptide FNQGQC1C2C3. This peptide contains the same adjacent active site cysteines. This latter peptide was subjected to LC-MS/MS, which enabled us to determine that the site of carbamoylation was at Cys2. The MS/MS product ion data also confirmed the presence of a carbamoyl group as the adduct species. PMID- 9416977 TI - Stereoselective inhibition of human butyrylcholinesterase by phosphonothiolate analogs of (+)- and (-)-cocaine. AB - The hydrolysis of cocaine (benzoylecgonine methyl ester) to ecgonine methyl ester by human butyrylcholinesterase (BuChE; EC 3.1.1.8) has been shown previously to constitute an important means to detoxicate this material to pharmacologically inactive metabolites. The naturally occurring (-)-cocaine is hydrolyzed to ecgonine methyl ester approximately 2000 times slower than the unnatural (+) cocaine isomer. In good agreement with previous studies, (-)-cocaine bound to human BuChE with relatively good affinity and competitively inhibited the hydrolysis of the spectrophotometric substrate butyrylthiocholine with a Ki value of 8.0 microM. Similarly, (+)-cocaine also showed relatively high affinity for the human BuChE and competitively inhibited butyrylthiocholine hydrolysis with a Ki value of 5.4 microM. The phosphonothiolates corresponding to the transition state analogs for both (-)- and (+)-cocaine hydrolysis were synthesized and tested as inhibitors of human BuChE-catalyzed hydrolysis of butyrylthiocholine. The phosphonothiolate corresponding to the transition state for (-)-cocaine hydrolysis was a competitive inhibitor with a Ki value of 55.8 microM. The phosphonothiolate corresponding to the transition state for (+)-cocaine hydrolysis gave a Ki value of 25.9 microM, but, in addition, it also showed irreversible inhibition with a ki of inactivation of 68.8 min-1 M-1. It is likely that the mechanism-based inhibitor described herein may find use as a mechanistic probe of butyrylcholinesterase action and also possibly aid in the purification of this class of esterases. PMID- 9416979 TI - Atrial fibrillation in hypertension: under recognised or over diagnosed? PMID- 9416978 TI - Interaction of brain cannabinoid receptors with guanine nucleotide binding protein: a radioligand binding study. AB - The binding of a classical cannabinoid agonist, [3H]R-(+)-(2,3-dihydro-5-methyl-3 [(4-morpholinyl)methyl]pyrol[1,2 ,3-de]-1,4-benzoxazin-6-yl)(1 napthalenyl)methanone monomethanesulfonate ([3H] WIN55212-2), and a selective cannabinoid receptor (CB1) antagonist, N-(piperidin-1-yl)-5-(4-chlorophenyl)1 (2,4-dichlorophenyl)-4-meth yl-1H-pyrazole-3-carboxamide hydrochloride ([3H]SR141716A), to rat cannabinoid receptors was evaluated using rat cerebellar membranes. Guanine nucleotides inhibited [3H]WIN55212-2 binding by approximately 50% at 10 microM and enhanced [3H]SR141716A binding very slightly. In the same tissue, the binding of guanosine 5'-O-[gamma-[35S]thio]triphosphate ([35S]GTP gamma-S) was characterized and the influence of cannabinomimetics evaluated on this binding. Cannabinoid receptor agonists enhanced [35S]GTP-gamma-S binding, whereas SR141716A was devoid of action by itself but antagonized the action of cannabinoid receptor agonists. The good correlation obtained between the half maximum efficient concentration (EC50) values in [35S]GTP-gamma-S binding and the IC50 values [3H]WIN55212-2 binding shows that [35S]GTP-gamma-S binding could be a good functional assay for brain cannabinoid receptors. PMID- 9416980 TI - Microalbuminuria in hypertension: is it up to measure? PMID- 9416981 TI - Management of atrial fibrillation in patients with hypertension. AB - Atrial fibrillation (AF) is a common arrhythmia in patients with hypertensive heart disease. In addition, the presence of hypertension in patients with AF constitutes an important risk factor for the development of thromboembolic events and probably also selects out those individuals who may be resistant to drug therapy. AF in patients with hypertensive heart disease may lead to a number of serious clinical sequelae including stroke, left atrial myopathy, left ventricular dysfunction, and congestive heart failure. This needs to be treated aggressively since many patients may become quite symptomatic when AF develops in the setting of diastolic and systolic dysfunction, regular features of hypertensive heart disease. There are several treatment approaches that may be considered in such patients ranging from interventions to prevent thromboembolic events, drugs and procedures for control of the ventricular response, and drug and non-pharmacologic therapy specifically designed to prevent AF or to restore normal sinus rhythm. This review article will cover each of these components of therapy of AF and will attempt to focus on those therapies that might be best suited for patients with hypertensive heart disease. PMID- 9416982 TI - Hypertension surveys in the developing world. Lessons from the Egyptian National Hypertension Project (NHP). AB - Accurate national estimates of the prevalence of hypertension in developing countries are lacking. Inadequate funds, inexperience and lack of infrastructure are also important barriers to hypertension research. The aim of this review is to help investigators from the developing countries, with limited resources, to design and conduct national hypertension surveys. The information is mostly based on the experience gained during the Egyptian National Hypertension Project (NHP) which can serve as a model for similar surveys elsewhere. The review addresses a number of important questions: (1) Why conduct a national hypertension survey in a developing country; (2) What kind of data are needed; (3) Where to start and how to raise funds; (4) Who will carry out the survey; (5) How to design your sample and where to survey; (6) How to organize and perform field operations; (7) How to collect accurate data and do quality control measures; and (8) How to handle the data? PMID- 9416983 TI - Microalbuminuria and its relation to cardiovascular disease and risk factors. A population-based study of 1254 hypertensive individuals. AB - Microalbuminuria has been proposed as a potential atherosclerotic risk factor in hypertensive individuals. The aim of this cross-sectional population study was to analyse whether microalbuminuria is related to a higher prevalence of cardiovascular disease, and a more atherogenic risk profile, and reversely related to the use of antihypertensive drugs. In a major health screening at the State University Hospital in Copenhagen, including urinary albumin excretion, glomerular filtration rate, blood pressure (BP), electrocardiogram, body mass index, plasma lipoproteins, fibrinogen, and albumin, and information regarding a history of acute myocardial infarction, smoking, and antihypertensive drugs, 1254 participants without diabetes mellitus or renal/urinary tract disease had arterial hypertension. Age range was 30-70 years. Microalbuminuria (nocturnal urinary albumin excretion >15 microg/min) occurred in 5%, and cardiovascular disease (previous acute myocardial infarction or electrocardiographic Q-waves) also in 5% of the study population. Microalbuminuric hypertensive subjects were characterized by higher age and systolic BP, and a male predominance, as compared to normoalbuminuric hypertensive subjects. The frequency of cardiovascular disease was similar in the two groups. In contrast, when analysed as a continuous variable, a one unit increase in the logarithmically transformed urinary albumin excretion significantly increased the likelihood of cardiovascular disease (odds ratio [95% confidence interval] 1.32 (1.02-1.70); P < 0.05), and this relation was independent of age, sex, and conventional atherosclerotic risk factors. Participants who were effectively treated with antihypertensive drugs did not have a lower urinary albumin excretion than insufficiently treated or untreated participants. It is concluded that slightly elevated albumin excretion in the urine is not only a pressure-dependent functional phenomenon in the glomerular vessel walls, but associated with permanent atherosclerotic abnormalities in the entire vascular system. PMID- 9416984 TI - Blood pressure levels in the 41 populations of the WHO MONICA Project. AB - In the early to mid 1980s, the WHO MONICA Project conducted cardiovascular risk factor surveys in 41 study populations in 22 countries. Study populations aged 35 64 years comprised 32,422 men and 32,554 women. Blood pressures (BP) and body mass index (BMI) were measured according to a standard protocol. Participants were asked about antihypertensive medication. In men, the average age standardized BPs ranged among the populations from 124 to 148 mm Hg for systolic (SBP) and from 75 to 93 mm Hg for diastolic (DBP). The corresponding values in women were 118-145 mm Hg for SBP and 74-90 mm Hg for DBP. In all populations, women had lower SBP than men in the age group 35-44. However, SBP in women rose more steeply with age so that in 34 of 41 populations women had higher SBP than men in the age group 55-64. The proportion of participants with untreated major elevation of BP ranged from 4.5% to 33.7% in men and from 1.9% to 22.3% in women. The proportions of participants receiving antihypertensive medication were 4.3 17.7% for men and 6.0-22.0% for women. These proportions were not correlated with the prevalence of untreated hypertensives. Age-adjusted BMI was associated with SBP and accounted for 14% of the SBP variance in men and 32% in women. We found a large difference in SBP among the MONICA study populations and conclude that the results represent a valid estimate of the public health problem posed by elevated BP. We also have shown that almost universally the problem of elevated BP is more prevalent in women than in men, especially in the older age groups. PMID- 9416985 TI - Risk of serious adverse events in hypertensive patients receiving isradipine: a meta-analysis. AB - A meta-analysis was performed to compare the risk of serious adverse events associated with the use of all formulations of isradipine, when used as monotherapy in hypertension, to active drug or placebo controls. Eligible studies totalled 65 published and unpublished randomised controlled trials involving 9903 subjects and 10,675 treatment exposures: 4492 to isradipine, 1473 to isradipine sustained release, 2768 to other active drugs, and 1942 to placebo. Mortality, cardiovascular outcomes, other serious incident illnesses, such as cancer, and withdrawals were sought. Seventy-five per cent of the isradipine exposures were to standard-release formulations and 25% were to sustained-release formulations. Overall, isradipine therapy shows no difference in risk of major adverse events or withdrawals compared to other active controls or placebo (odds ratios [OR] 0.9; 95% CI 0.7-1.46 and 0.5; 95% CI 0.2-1.3). These major adverse events included angina, fatal and non-fatal myocardial infarction, stroke and overall mortality. Isradipine sustained release could be compared only to placebo, based on available data, and shows a lower risk of withdrawals (OR 0.5; 95% CI 0.3 0.9), and a similar trend was observed for major adverse events, (OR 0.8; 95% CI 0.3-2.5). Published and unpublished randomised controlled trials were analysed in separate meta-analyses and later combined when this sensitivity analysis of risk showed no differences between the groups. In conclusion, we find no evidence for increased risk of serious adverse events in patients receiving isradipine as monotherapy for hypertension. PMID- 9416986 TI - Comparative effects of nebivolol and atenolol on blood pressure and insulin sensitivity in hypertensive subjects with type II diabetes. AB - The aim of this double-blind, parallel group study was to compare the effects of nebivolol and atenolol on blood pressure (BP) and insulin sensitivity in hypertensive patients with type II, non-insulin dependent diabetes mellitus (NIDDM). After a 4-week run-in period on placebo, 30 patients (14 males and 16 females) aged 43 to 69 years, with stable NIDDM and mild to moderate hypertension (DBP > or =95 and <116 mm Hg) were randomised to receive either nebivolol 5 mg or atenolol 50 mg, both administered once daily for 6 months. At the end of the placebo and the active treatment periods, supine and standing BP was measured, 24 h urinary C-peptide, HbA1c, plasma glucose and lipid levels were evaluated and an euglycaemic hyperinsulinaemic clamp was performed to evaluate insulin sensitivity: glucose infusion rate during the last 60 min of clamp and total glucose requirements were evaluated. Nebivolol 5 mg once daily was of an equivalent efficacy as atenolol 50 mg once daily at reducing supine and standing systolic and diastolic BP values. Neither beta-blocker adversely affected carbohydrate metabolism in terms of insulin sensitivity, whole body glucose utilization, HbA1c and 24-h urinary C-peptide excretion. No significant changes in cholesterol (total, high density and low density lipoprotein) and triglycerides plasma levels were observed with both beta-blockers. These findings indicate that, in hypertensive patients with NIDDM, ie, in subjects who have established insulin resistance, treatment with nebivolol and atenolol neither further deteriorated insulin sensitivity nor adversely affected the lipid profile. PMID- 9416987 TI - A low dose atenolol/bendrofluazide combination in patients with mild to moderate hypertension. PMID- 9416988 TI - PACAP-induced plasma extravasation in rat skin. AB - The effects of pituitary adenylate cyclase activating peptide (PACAP) 38, PACAP 27 and vasoactive intestinal peptide (VIP) on plasma extravasation were investigated in vivo in rat skin. PACAP 38, PACAP 27 and VIP, caused concentration-dependent extravasation in rat skin. The order of potency was PACAP 38 > PACAP 27 = VIP, whereas the order of maximal induced extravasation was PACAP 38 = PACAP 27 > VIP, suggesting that PACAP 38 might be the most powerful inducer of plasma extravasation of the three tested members of the secretin-glucagon-VIP family. Substance P (SP) was about 5 times more potent than PACAP 38 and 15 times more potent than PACAP 27. These data indicate that PACAP 38 induced plasma extravasation in concentrations roughly equimolar to SP. Pyrilamine (H1 receptor antagonist) reduced the PACAP 38-induced plasma extravasation more than 50%; cimetidine (H2 receptor antagonist) was without effect. To investigate whether a cAMP-mediated process is involved in the induction of plasma extravasation, the synthetic adenosine 3',5'-cyclic monophosphate (cAMP), dibutyryl adenosine cyclic monophosphate (DBcAMP) and the cAMP-inducing drug, salbutamol, were each injected in the skin; neither of these drugs caused extravasation. We conclude that PACAP 38 and PACAP 27 cause potent plasma extravasation which, at least in part, involves histamine release. PMID- 9416989 TI - Neurohormonal regulation of histamine and pancreastatin secretion from isolated rat stomach ECL cells. AB - ECL cells are numerous in the acid-producing part of the rat stomach. They are rich in histamine and pancreastatin, a chromogranin A-derived peptide, and they secrete these products in response to gastrin. We have examined how isolated ECL cells respond to a variety of neuromessengers and peptide hormones. Highly purified (85%) ECL cells were collected from rat stomach using repeated counter flow elutriation and cultured for 48 h before experiments were conducted. The ECL cells responded to gastrin, sulphated cholecystokinin-8 and to high K+ and Ca2+ with the parallel secretion of histamine and pancreastatin. Glycine-extended gastrin was without effect. Forskolin, an activator of adenylate cyclase, induced secretion, whereas isobutylmethylxanthine, a phosphodiesterase inhibitor, raised the basal release without enhancing the gastrin-evoked stimulation. Maximum stimulation with gastrin resulted in the release of 30% of the secretory products. Numerous neuromessengers and peptide hormones were screened for their ability to stimulate secretion and to inhibit gastrin-stimulated secretion. Pituitary adenylate cyclase activating peptide (PACAP)-27 and -38 stimulated secretion of both histamine and pancreastatin with a potency greater than that of gastrin and with the same efficacy. Related peptides, such as vasoactive intestinal peptide, helodermin and helospectin, stimulated secretion with lower potency. The combination of EC100 gastrin and EC50 PACAP produced a greater response than gastrin alone. None of the other neuropeptides or peptide hormones tested stimulated secretion. Serotonin, adrenaline, noradrenaline and isoprenaline induced moderate secretion at high concentrations. Muscarinic receptor agonists did not stimulate secretion, and histamine and selective histamine receptor agonists and antagonists were without effect. This was the case also with GABA, aspartate and glutamate. Somatostatin and galanin, but none of the other agents tested, inhibited gastrin-stimulated secretion. Our results reveal that not only gastrin but also PACAP is a powerful excitant of the ECL cells, that not only somatostatin, but also galanin can suppress secretion, that muscarinic receptor agonists fail to evoke secretion, and that histamine (and pancreastatin) does not evoke autofeedback inhibition. PMID- 9416990 TI - Functional characterization of neural and smooth muscle motilin receptors in the chicken proventriculus and ileum. AB - To characterize the motilin receptors present in the chicken, the effects of chicken motilin (Phe-Val-Pro-Phe-Phe-Thr-Gln-Ser-Asp-Ile-Gln-Lys-Met-Gln-Glu-Lys Glu-Arg -Asn-Lys-Gly-Gln), Leu13 porcine motilin, canine motilin and three erythromycin derivatives (EMA, EM523, GM611) on the contractility of the chicken gastrointestinal (GI) smooth muscles were investigated in vitro and compared with those in the rabbit duodenum. In the proventriculus longitudinal and circular muscle layers, chicken motilin (3 nM-1 microM) caused an atropine- and a tetrodotoxin-sensitive contraction (EC50 = 39-49 nM), and potentiated the EFS induced contraction without affecting the responsiveness of acetylcholine. EM523 and GM611 (3-100 microM) contracted the proventriculus longitudinal muscle, and the maximum amplitudes of contraction were about 60% of that induced by chicken motilin. Chicken motilin (0.1 nM-100 nM) also caused contraction of the ileum (EC50 = 7 nM) through direct action on the smooth muscle cells. On the other hand, erythromycin derivatives showed only a weak contractile efficacy (about 20% of the maximum response of chicken motilin) even at high concentrations (10-100 microM). The rank order of potency in the ileum was chicken motilin > canine motilin > or = Leu13 porcine motilin > > GM611 > or = EM523 > or = EMA. GM109 slightly inhibited the ideal contractions induced by Leu13 porcine motilin at 100 microM (pA2 = 3.86). In the rabbit duodenum, chicken motilin was a full agonist with the same intrinsic activity as Leu13 porcine motilin, canine motilin and the erythromycin derivatives. However, the rank order of potency (Leu13 porcine motilin > or = canine motilin > chicken motilin > GM611 > or = EM523 > EMA) was different from that in the chicken ileum. In conclusion, chicken motilin causes an excitatory response in the chicken GI tract through activation of neural (proventriculus) and smooth muscle motilin receptors (ileum). The motilin receptor present in the ileum is different from that demonstrated in the rabbit intestine, because of a different rank order of motilin peptides in producing the contraction, low contracting activity of erythromycin derivatives and low antagonistic efficacy of GM109. Different pharmacological characteristics of the mechanical response induced by motilin peptides and erythromycin derivatives between the proventriculus and the ileum are discussed. PMID- 9416991 TI - Characterization of the receptor response for the neuropeptide Y-evoked suppression of parasympathetically-mediated contractions in the guinea pig trachea. AB - Neuropeptide Y (NPY) acts via several distinct receptor types. The aim of the present study was to examine which NPY receptors are coupled to inhibition of parasympathetically-mediated contractions of the isolated guinea pig trachea. Electrical field stimulation of tracheal rings evoked a rapid twitch, which was abolished by atropine (1 microM). NPY, the structurally related hormone peptide YY (PYY), the Y2 receptor agonist [Cys2, Aoc5-24, D-Cys27]NPY, as well as NPY 5 36 and NPY 13-36 evoked a concentration-dependent inhibition of the electrically stimulated twitches. Pretreatment with the Y1 receptor-selective antagonist BIBP3226 (1 microM) failed to prevent the NPY-induced inhibition. Although less potent than NPY, the Y1 (and Y4-Y6) receptor agonist [Leu31, Pro34]NPY also inhibited the electrically-stimulated twitches. Another NPY-related peptide, pancreatic polypeptide, which recognizes Y4-Y6 receptors did not affect the stimulated twitches at concentrations up to 1 microM. However, pretreatment with the Y1 receptor-selective antagonist BIBP3226 (1 microM) virtually abolished the inhibition evoked by [Leu31, Pro34]NPY. None of the peptides affected the baseline tension and BIBP3226 (1 microM) per se did not affect the amplitude of the electrically-stimulated twitches. In conclusion, it seems that NPY and PYY are capable of suppressing parasympathetically mediated contractions in the guinea pig trachea mainly via Y2 receptors, but there is also a small contribution from Y1 receptors. PMID- 9416992 TI - Holiday lectures bring tidings of good science. PMID- 9416994 TI - Momentum grows behind plans for health museum. PMID- 9416993 TI - CT scans probe secrets of Italian masters' violins. PMID- 9416995 TI - From the Centers for Disease Control and Prevention. Update: perinatally acquired HIV/AIDS--United States, 1997. PMID- 9416996 TI - A piece of my mind. Damascus Road. PMID- 9416997 TI - Patient consent for publication. PMID- 9416998 TI - Patient consent for publication. PMID- 9416999 TI - Patient consent for publication. PMID- 9417000 TI - Brain serotonin neurotoxicity and fenfluramine and dexfenfluramine. PMID- 9417001 TI - Brain serotonin neurotoxicity and fenfluramine and dexfenfluramine. PMID- 9417002 TI - Brain serotonin neurotoxicity and fenfluramine and dexfenfluramine. PMID- 9417003 TI - Brain serotonin neurotoxicity and fenfluramine and dexfenfluramine. PMID- 9417004 TI - APOE epsilon4 allele and chronic traumatic brain injury. PMID- 9417005 TI - False-negative qualitative cardiac troponin T in a 79-year-old man with myocardial infarction. PMID- 9417006 TI - Elevation of troponin I levels in patients without evidence of myocardial injury. PMID- 9417007 TI - Inverse association of dietary fat with development of ischemic stroke in men. AB - CONTEXT: A few ecological and cohort studies in Asian populations suggest an inverse association of the intake of both fat and saturated fat with risk of stroke. However, data among western populations are scant. OBJECTIVE: To examine the association of stroke incidence with intake of fat and type of fat among middle-aged US men during 20 years of follow-up. DESIGN AND SETTING: The Framingham Heart Study, a population-based cohort study. PARTICIPANTS: A total of 832 men, aged 45 through 65 years, who were free of cardiovascular disease at baseline (1966-1969). MEASUREMENTS AND DATA ANALYSIS: The diet of each subject was assessed at baseline by a single 24-hour dietary recall, from which intakes of energy and macronutrients were estimated. In Kaplan-Meier analyses, we calculated age-adjusted cumulative incidence rates of stroke. Using Cox regression, we estimated stroke incidence relative risks during 20 years of follow-up. MAIN OUTCOME MEASURE: Incidence of ischemic stroke, which occurred in 61 subjects during the follow-up period. RESULTS: Mean intakes were 10975 kJ for energy; 114 g (39% of energy) for total fat; 44 g (15%) for saturated fat; 46 g (16%) for monounsaturated fat; and 16 g (5%) for polyunsaturated fat. Risk of ischemic stroke declined across the increasing quintile of total fat (log-rank trend P=.008), saturated fat (P=.002), and monounsaturated fat (P=.008) but not polyunsaturated fat (P=.33). The age- and energy-adjusted relative risk for each increment of 3% of energy from total fat was 0.85 (95% confidence interval [CI], 0.78-0.94); for an increment of 1% from saturated fat, 0.91 (95% CI, 0.85-0.98); and for 1% from monounsaturated fat, 0.89 (95% CI, 0.83-0.96). Adjustment for cigarette smoking, glucose intolerance, body mass index, blood pressure, blood cholesterol level, physical activity, and intake of vegetables and fruits and alcohol did not materially change the results. Too few cases of hemorrhagic stroke (n=14) occurred to draw inferences. CONCLUSION: Intakes of fat, saturated fat, and monounsaturated fat were associated with reduced risk of ischemic stroke in men. PMID- 9417008 TI - Prospective evaluation of a clinical guideline recommending hospital length of stay in upper gastrointestinal tract hemorrhage. AB - CONTEXT: Upper gastrointestinal tract hemorrhage (UGIH) is a common and potentially life-threatening disorder. Resource utilization can vary without adverse effect on patient outcome. Clinical practice guidelines are a potential solution to reduce variation in practice while improving patient outcomes. OBJECTIVE: To validate prospectively the safety, acceptability, and impact of a clinical practice guideline defining the medically appropriate length of stay (LOS) for patients hospitalized with UGIH. DESIGN: Prospective, controlled time series study with an alternate-month design. Outcome surveyors and patients were blinded to study group allocation. GUIDELINE: A retrospectively validated scoring system using 4 independent variables: hemodynamics, time from bleeding, comorbidity, and esophagogastroduodenoscopy (EGD) findings to predict risk of adverse events. The quantitative risk for the low-risk subset was 0.6% (95% confidence interval [CI], 0.0%-2.0%) for subsequent complications and 0% (95% CI, 0.0%-0.9%) for life-threatening complications from this retrospective evaluation. SETTING: A 1000-bed, not-for-profit, university-affiliated teaching hospital. PATIENTS: Consecutive adult patients hospitalized for acute UGIH. INTERVENTION: Concurrent feedback of guideline recommendation (same-day hospital discharge) to physicians caring for patients at low risk for complication. No risk information was provided during control months. RESULTS: Seventy percent (209/299) of UGIH patients achieved low-risk status according to the guideline and were therefore potentially suitable for early discharge from the hospital. Providing real-time quantitative risk information (intervention group only) was associated with an increase in guideline compliance from 30% to 70% (P<.001) and a decrease in mean (SD) LOS from 4.6 (3.5) days to 2.9 (1.3) days (mean reduction of 1.7 days per patient; P<.001). No differences in complications, patient health status, or patient satisfaction were found when measured 1 month after discharge. An independent variable predicting decreased hospital LOS for low-risk UGIH patients was early EGD. CONCLUSIONS: Implementation of the clinical practice guideline safely reduced hospital LOS for selected low-risk patients with acute UGIH. Further prospective validation in other settings is warranted. PMID- 9417009 TI - Comparison of ring block, dorsal penile nerve block, and topical anesthesia for neonatal circumcision: a randomized controlled trial. AB - CONTEXT: Beliefs about the safety and effectiveness of current anesthetics have resulted in many newborns being circumcised without the benefit of anesthesia. OBJECTIVE: To compare ring block, dorsal penile nerve block, a topical eutectic mixture of local anesthetics (EMLA), and topical placebo when used for neonatal circumcision. The placebo represented current practice, with no anesthetic for neonatal circumcision. DESIGN: A randomized controlled trial. SETTING: Antenatal units in 2 tertiary care hospitals in Edmonton, Alberta. PARTICIPANTS: A consecutive sample of 52 healthy, full-term, male newborns, aged 1 to 3 days. INTERVENTIONS: Physiological and behavioral monitoring occurred in a series of trials: baseline, drug application, preparation, circumcision, and postcircumcision. Surgical procedures defined the following 4 stages of the circumcision: cleansing, separation, clamp on, and clamp off. Methemoglobin level was assessed 6 hours after surgery. MAIN OUTCOME MEASURES: Heart rate, cry, and methemoglobin level. RESULTS: Newborns in the untreated placebo group exhibited homogeneous responses that consisted of sustained elevation of heart rate and high-pitched cry throughout the circumcision and following. Two newborns in the placebo group became ill following circumcision (choking and apnea). The 3 treatment groups all had significantly less crying and lower heart rates during and following circumcision compared with the untreated group. The ring block was equally effective through all stages of the circumcision, whereas the dorsal penile nerve block and EMLA were not effective during foreskin separation and incision. Methemoglobin levels were highest in the EMLA group, although no newborn required treatment. CONCLUSIONS: The most effective anesthetic is the ring block; EMLA is the least effective. It is our recommendation that an anesthetic should be administered to newborns prior to undergoing circumcision. PMID- 9417010 TI - Occupational injuries among workers with disabilities: the National Health Interview Survey, 1985-1994. AB - CONTEXT: As the baby boom generation ages, more people will be working with disabilities, but we have little information regarding how disabilities affect risk for occupational injury. OBJECTIVE: To test the hypothesis that work limiting disabilities in general and hearing and visual impairments in particular are risk factors for occupational injuries. DESIGN: Retrospective cohort study. SETTING: The National Health Interview Survey (NHIS), 1985 to 1994. PARTICIPANTS: The 459827 participants in the NHIS from 1985 to 1994 who listed "working" as their primary activity, who were not farmers, and who were between 18 and 65 years of age. MAIN OUTCOME MEASURE: Occupational injuries in the year preceding the interview causing a residual impairment at the time of interview. RESULTS: After adjusting for occupation, self-employment, and age, occupational injury was associated with preceding work disability (odds ratio [OR], 1.36; 95% confidence interval [CI], 1.19-1.56); blindness (OR, 3.21; 95% CI, 1.32-7.85); deafness (OR, 2.19; 95% CI, 1.17-4.12); hearing impairment (OR,1.55; 95% CI, 1.29-1.87); upper extremity impairment (OR, 1.46; 95% CI, 1.05-2.05); and arthritis (OR, 1.34; 95% CI, 1.07-1.68). Visual impairment was not associated with a significantly increased risk (OR, 1.37; 95% CI, 0.87-2.17). CONCLUSIONS: Workers with disabilities, especially sensory impairments, appear to have an elevated risk for occupational injury. Further research in the design and evaluation of improved workplace accommodations for workers with these disabilities is needed. PMID- 9417011 TI - Hepatitis B virus transmission in an elementary school setting. AB - CONTEXT: The risk of transmission of hepatitis B virus (HBV) in day care centers and schools is low. OBJECTIVE: To investigate the source of HBV transmission for an elementary school teacher with acute hepatitis B. DESIGN: Serologic survey for HBV infection among elementary school students, school staff, and household members of an HBV-infected teacher and student. SETTING: General community and elementary school. PATIENTS: Elementary school students and staff members and household members of an HBV-infected teacher. MAIN OUTCOME MEASURES: Elementary school students, school staff, and household members of an HBV-infected teacher were tested for markers of HBV infection. Samples positive for hepatitis B surface antigen (HBsAg) were tested for HBsAg subtype using monoclonal antibodies and examined for HBV DNA homology by polymerase chain reaction techniques. RESULTS: An HBV-infected student and the teacher were found to have the same HBV subtype (ayw1-2) and to have identical HBV DNA sequences. The teacher reported none of the usual risk factors for acquiring HBV infection, and none of her family members had been infected prior to her illness. The specific means of HBV transmission from student to teacher was not identified. Of 108 total children in the same grade as the HBV-infected student, 102 (94%) were tested for serologic markers of HBV infection, and none was positive. CONCLUSIONS: This investigation documented transmission from an HBV-infected student to a teacher in an elementary school setting without a reported overt percutaneous or permucosal exposure to blood or infectious body fluids. Transmission of HBV to other students or staff members in the school was not observed. PMID- 9417012 TI - Benzodiazepines and zolpidem for chronic insomnia: a meta-analysis of treatment efficacy. AB - OBJECTIVE: To evaluate the efficacy of benzodiazepines and zolpidem tartrate in chronic insomnia based on a quantitative review of literature. DATA SOURCES: Articles from 1966 to 1996 were identified using MEDLINE, by a manual review of relevant journals, and from bibliographies of identified articles. STUDY SELECTION: Studies using randomized, double-blind, placebo-controlled, parallel or crossover designs with benzodiazepines or zolpidem in adults younger than 65 years with chronic insomnia (modified Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria for primary insomnia) were selected for review. Self-report and polysomnographic measures of sleep-onset latency, total sleep time, number of awakenings, and sleep quality were selected as outcomes. DATA EXTRACTION: Twenty-two studies met the selection criteria. A combined test of Pvalues was performed, pooling broadly from the 22 studies to determine whether medication was superior to placebo. A combined test of effect sizes was performed on the subset of studies that reported effect size information to determine the magnitude of medication effect. DATA SYNTHESIS: A homogeneous sample of studies summarized 1894 patients treated for a median duration of 7 days. The combined test of P values demonstrated that medication was superior to placebo in all 4 outcome measures. Treatment response was moderate in magnitude by the combined test of effect sizes. CONCLUSIONS: Benzodiazepines and zolpidem produced reliable improvements in commonly measured parameters of sleep in patients with chronic insomnia. Relative to the chronic and recurring course of insomnia, both the limited duration of treatments studied and the lack of follow up data from controlled trials represent challenges for developing evidence-based guidelines for the use of hypnotics in the management of chronic insomnia. PMID- 9417013 TI - A 73-year-old man with symptomatic benign prostatic hyperplasia. PMID- 9417014 TI - An 88-year-old woman facing the end of life, 1 year later. PMID- 9417015 TI - Fat chance: diet and ischemic stroke. PMID- 9417016 TI - Using a practice guideline for safely shortening hospital stay for upper gastrointestinal tract hemorrhage. PMID- 9417017 TI - Ultraviolet-induced cell death blocked by a selenoprotein from a human dermatotropic poxvirus. AB - Selenium, an essential trace element, is a component of prokaryotic and eukaryotic antioxidant proteins. A candidate selenoprotein homologous to glutathione peroxidase was deduced from the sequence of molluscum contagiosum, a poxvirus that causes persistent skin neoplasms in children and acquired immunodeficiency syndrome (AIDS) patients. Selenium was incorporated into this protein during biosynthesis, and a characteristic stem-loop structure near the end of the messenger RNA was required for alternative selenocysteine decoding of a potential UGA stop codon within the open reading frame. The selenoprotein protected human keratinocytes against cytotoxic effects of ultraviolet irradiation and hydrogen peroxide, providing a mechanism for a virus to defend itself against environmental stress. PMID- 9417018 TI - Change in chemoattractant responsiveness of developing axons at an intermediate target. AB - Developing axons reach their final targets as a result of a series of axonal projections to successive intermediate targets. Long-range chemoattraction by intermediate targets plays a key role in this process. Growing axons, however, do not stall at the intermediate targets, where the chemoattractant concentration is expected to be maximal. Commissural axons in the metencephalon, initially attracted by a chemoattractant released from the floor plate, were shown to lose responsiveness to the chemoattractant when they crossed the floor plate in vitro. Such changes in axon responsiveness to chemoattractants may enable developing axons to continue to navigate toward their final destinations. PMID- 9417019 TI - Hydrogen radicals, nitrogen radicals, and the production of O3 in the upper troposphere AB - The concentrations of the hydrogen radicals OH and HO2 in the middle and upper troposphere were measured simultaneously with those of NO, O3, CO, H2O, CH4, non methane hydrocarbons, and with the ultraviolet and visible radiation field. The data allow a direct examination of the processes that produce O3 in this region of the atmosphere. Comparison of the measured concentrations of OH and HO2 with calculations based on their production from water vapor, ozone, and methane demonstrate that these sources are insufficient to explain the observed radical concentrations in the upper troposphere. The photolysis of carbonyl and peroxide compounds transported to this region from the lower troposphere may provide the source of HOx required to sustain the measured abundances of these radical species. The mechanism by which NO affects the production of O3 is also illustrated by the measurements. In the upper tropospheric air masses sampled, the production rate for ozone (determined from the measured concentrations of HO2 and NO) is calculated to be about 1 part per billion by volume each day. This production rate is faster than previously thought and implies that anthropogenic activities that add NO to the upper troposphere, such as biomass burning and aviation, will lead to production of more O3 than expected. PMID- 9417020 TI - Infrared photorefractive polymers and their applications for imaging. AB - Photorefractive polymers with high diffraction efficiency in the visible and near infrared regions of the electromagnetic spectrum have been developed. These polymers, which have a large dynamic range because of their high orientational birefringence, incorporate a dye designed to have a large dipole moment and a high linear polarizability anisotropy. Such polymers have enabled demonstrations of imaging through scattering media, using a holographic time-gating technique at a wavelength that is compatible with the transparency of biological tissues and with the emission of low-cost semiconductor laser diodes. PMID- 9417021 TI - Electromechanical properties of an ultrathin layer of directionally aligned helical polypeptides. AB - The electromechanical properties of a monomolecular film of poly-gamma-benzyl-L glutamate (PBLG) 15 nanometers thick grafted at the carboxyl-terminal end to a flat aluminum surface were measured. The field-induced change in film thickness, dominated by a large inverse-piezoelectric effect, demonstrates that the "grafting-from" technique forces the chains into a parallel arrangement. The mechanical plate modulus of the film as determined by electrostriction agrees with the theoretical prediction for a single PBLG molecule along the chain axis. The experiments show that ultrathin polypeptide layers with large persistent polarization can be fabricated by the grafting approach. PMID- 9417022 TI - Coupling of south american and african plate motion and plate deformation AB - Although the African Plate's northeastward absolute motion slowed abruptly 30 million years ago, the South Atlantic's spreading velocity has remained roughly constant over the past 80 million years, thus requiring a simultaneous westward acceleration of the South American Plate. This plate velocity correlation occurs because the two plates are coupled to general mantle circulation. The deceleration of the African Plate, due to its collision with the Eurasian Plate, diverts mantle flow westward, increasing the net basal driving torque and westward velocity of the South American Plate. One result of South America's higher plate velocity is the increased cordilleran activity along its western edge, beginning at about 30 million years ago. PMID- 9417023 TI - Footwall refrigeration along a detachment fault: implications for the thermal evolution of core complexes AB - Oxygen isotope compositions of epidote and quartz from chloritic breccias that underlie the detachment fault in the metamorphic core complex of the Whipple Mountains yielded quartz-epidote fractionations that range from 4.1 to 6.4 per mil and increase systematically toward the fault. These fractionations give mean temperatures that decrease from approximately 432 degrees C at 50 meters below the fault to approximately 350 degrees C at 12 meters below the fault. This extreme thermal gradient of 82 degrees C over 38 meters (2160 degrees C per kilometer) is best explained by advective heat extraction by means of circulating surface-derived fluids. Models of lithospheric extension consider only conductive cooling resulting from tectonic denudation and thus require revision to include fluid-induced fault-zone refrigeration. PMID- 9417024 TI - Suppression of volcanism during rapid extension in the basin and range province, united states AB - Continental extension and volcanism are generally thought to be complementary. Stratigraphic and structural data from some highly extended parts of the Basin and Range province reveal instead that rapid extension appears to have suppressed volcanism. This relation may reflect enhanced crystallization of midcrustal magmas during extension resulting from exsolution of magmatic volatiles, increased interaction of magmas with meteoric water, and dispersal of magma into smaller bodies. Some rift environments may thus be characterized by voluminous synextensional plutonism with little or no concomitant volcanism. PMID- 9417025 TI - Migrating planets AB - A planet orbiting in a disk of planetesimals can experience an instability in which it migrates to smaller orbital radii. Resonant interactions between the planet and planetesimals remove angular momentum from the planetesimals, increasing their eccentricities. Subsequently, the planetesimals either collide with or are ejected by the planet, reducing the semimajor axis of the planet. If the surface density of the planetesimals exceeds a critical value, corresponding to approximately 0.03 solar mass of gas inside the orbit of Jupiter, the planet will migrate inward a large distance. This instability may explain the presence of Jupiter-mass objects in small orbits around nearby stars. PMID- 9417026 TI - Magnetic properties of hexagonal closed-packed iron deduced from direct observations in a diamond anvil cell AB - The attraction of hexagonal closed packed (hcp) iron to a magnet at 16.9 gigapascals and 261 degrees centigrade suggests that hcp iron is either paramagnetic or ferromagnetic with susceptibilities from 0. 15 to 0.001 and magnetizations from 1800 to 15 amperes per meter. If dominant in Earth's inner core, paramagnetic hcp iron could stabilize the geodynamo. PMID- 9417027 TI - Formation of molecular chlorine from the photolysis of ozone and aqueous sea-salt particles AB - Halogen atoms from the reactions of sea-salt particles may play a significant role in the marine boundary layer. Reactions of sodium chloride, the major component of sea-salt particles, with nitrogen oxides generate chlorine atom precursors. However, recent studies suggest there is an additional source of chlorine in the marine troposphere. This study shows that molecular chlorine is generated from the photolysis of ozone in the presence of sea-salt particles above their deliquescence point; this process may also occur in the ocean surface layer. Given the global distribution of ozone, this process may provide a global source of chlorine. PMID- 9417028 TI - Broad-spectrum, non-opioid analgesic activity by selective modulation of neuronal nicotinic acetylcholine receptors. AB - Development of analgesic agents for the treatment of severe pain requires the identification of compounds that are devoid of opioid receptor liabilities. A potent (inhibition constant = 37 picomolar) neuronal nicotinic acetylcholine receptor (nAChR) ligand called ABT-594 was developed that has antinociceptive properties equal in efficacy to those of morphine across a series of diverse animal models of acute thermal, persistent chemical, and neuropathic pain states. These effects were blocked by the nAChR antagonist mecamylamine. In contrast to morphine, repeated treatment with ABT-594 did not appear to elicit opioid-like withdrawal or physical dependence. Thus, ABT-594 may be an analgesic that lacks the problems associated with opioid analgesia. PMID- 9417029 TI - Inhibition of the hammerhead ribozyme cleavage reaction by site-specific binding of Tb. AB - Terbium(III) [Tb(III)] was shown to inhibit the hammerhead ribozyme by competing with a single magnesium(II) ion. X-ray crystallography revealed that the Tb(III) ion binds to a site adjacent to an essential guanosine in the catalytic core of the ribozyme, approximately 10 angstroms from the cleavage site. Synthetic modifications near this binding site yielded an RNA substrate that was resistant to Tb(III) binding and capable of being cleaved, even in the presence of up to 20 micromolar Tb(III). It is suggested that the magnesium(II) ion thought to bind at this site may act as a switch, affecting the conformational changes required to achieve the transition state. PMID- 9417030 TI - Quantitation of transcription and clonal selection of single living cells with beta-lactamase as reporter. AB - Gene expression was visualized in single living mammalian cells with beta lactamase as a reporter that hydrolyzes a substrate loaded intracellularly as a membrane-permeant ester. Each enzyme molecule changed the fluorescence of many substrate molecules from green to blue by disrupting resonance energy transfer. This wavelength shift was detectable by eye or color film in individual cells containing less than 100 beta-lactamase molecules. The robust change in emission ratio reveals quantitative heterogeneity in real-time gene expression, enables clonal selection by flow cytometry, and forms a basis for high-throughput screening of pharmaceutical candidate drugs in living mammalian cells. PMID- 9417031 TI - Dimerization-induced inhibition of receptor protein tyrosine phosphatase function through an inhibitory wedge. AB - The function and regulation of the receptorlike transmembrane protein tyrosine phosphatases (RPTPs) are not well understood. Ligand-induced dimerization inhibited the function of the epidermal growth factor receptor (EGFR)-RPTP CD45 chimera (EGFR-CD45) in T cell signal transduction. Properties of mutated EGFR CD45 chimeras supported a general model for the regulation of RPTPs, derived from the crystal structure of the RPTPalpha membrane-proximal phosphatase domain. The phosphatase domain apparently forms a symmetrical dimer in which the catalytic site of one molecule is blocked by specific contacts with a wedge from the other. PMID- 9417032 TI - Dissociated pattern of activity in visual cortices and their projections during human rapid eye movement sleep. AB - Positron emission tomography was used to measure cerebral activity and to evaluate regional interrelationships within visual cortices and their projections during rapid eye movement (REM) sleep in human subjects. REM sleep was associated with selective activation of extrastriate visual cortices, particularly within the ventral processing stream, and an unexpected attenuation of activity in the primary visual cortex; increases in regional cerebral blood flow in extrastriate areas were significantly correlated with decreases in the striate cortex. Extrastriate activity was also associated with concomitant activation of limbic and paralimbic regions, but with a marked reduction of activity in frontal association areas including lateral orbital and dorsolateral prefrontal cortices. This pattern suggests a model for brain mechanisms subserving REM sleep where visual association cortices and their paralimbic projections may operate as a closed system dissociated from the regions at either end of the visual hierarchy that mediate interactions with the external world. PMID- 9417033 TI - Discrete start sites for DNA synthesis in the yeast ARS1 origin. AB - Sites of DNA synthesis initiation have been detected at the nucleotide level in a yeast origin of bidirectional replication with the use of replication initiation point mapping. The ARS1 origin of Saccharomyces cerevisiae showed a transition from discontinuous to continuous DNA synthesis in an 18-base pair region (nucleotides 828 to 845) from within element B1 toward B2, adjacent to the binding site for the origin recognition complex, the putative initiator protein. PMID- 9417034 TI - Modification of the NADH of the isoniazid target (InhA) from Mycobacterium tuberculosis. AB - The preferred antitubercular drug isoniazid specifically targets a long-chain enoyl-acyl carrier protein reductase (InhA), an enzyme essential for mycolic acid biosynthesis in Mycobacterium tuberculosis. Despite the widespread use of this drug for more than 40 years, its precise mode of action has remained obscure. Data from x-ray crystallography and mass spectrometry reveal that the mechanism of isoniazid action against InhA is covalent attachment of the activated form of the drug to the nicotinamide ring of nicotinamide adenine dinucleotide bound within the active site of InhA. PMID- 9417035 TI - The p53 network. PMID- 9417036 TI - Uncoupling protein-3 expression in rodent skeletal muscle is modulated by food intake but not by changes in environmental temperature. AB - A new member of the uncoupling protein (UCP) family called UCP3 has recently been cloned and shown to be highly expressed in skeletal muscle of rodents and humans. In the present study, UCP3 was overexpressed in C2C12 myoblasts where it acts as an uncoupling protein. Changes in UCP3 mRNA expression were examined in rodent muscles under conditions known to modulate thermogenesis in brown adipose tissue. In skeletal muscle, UCP3 expression did not change in response to 48 h of cold exposure (6 degrees C), whereas it was decreased by 81% or increased 5.6-fold by 1 week of 50% food restriction or fasting, respectively. It was also decreased by 36% in soleus muscle of obese (fa/fa) as compared with lean Zucker rats. The unexpected rise of UCP3 mRNA level induced by fasting did not change in vitro muscle basal heat production rate but decreased by 31% the capacity to produce heat in response to the uncoupler carbonylcyanide p trifluoromethoxyphenylhydrazone. This decrease may reflect underlying uncoupling by UCP3. Up-regulation of UCP3 mRNA after a 24-h fast was still observed in mice exposed at thermoneutrality. These results show that the increase in UCP3 expression induced by fasting is associated with the maintenance of thermogenesis measured in muscle in vitro and is not modulated by environmental temperature. The notion that UCP3 expression is modulated by food intake is of importance to better understand the pathophysiology of obesity in humans. PMID- 9417037 TI - Activation of beta1 integrin signaling stimulates tyrosine phosphorylation of p190RhoGAP and membrane-protrusive activities at invadopodia. AB - The ligation of available alpha6beta1 integrin in adherent LOX melanoma cells by laminin G peptides and integrin stimulatory antibodies induced cell invasiveness, independent of adhesion activity of integrins that were pre-bound to extracellular matrix (Nakahara, H., Nomizu, M., Akiyama, S. K., Yamada, Y., Yeh, Y., and Chen, W.-T. (1996) J. Biol. Chem. 271, 27221-27224). Here, we show that this induced invasion involves an increase in tyrosine phosphorylation of a 190 kDa GTPase-activating protein for Rho family members (p190(RhoGAP); p190) and membrane-protrusive activities at invadopodia. This tyrosine phosphorylation does not occur when the adherent cells are treated with non-activating antibody against beta1 integrin, control laminin peptides, or tyrosine kinase inhibitors genistein and herbimycin A. Although p190 and F-actin co-distribute in all cell cortex extensions, tyrosine-phosphorylated proteins including p190 appear to associate with F-actin specifically in invadopodia. In addition, the localized matrix degradation and membrane-protrusive activities were blocked by treatment of LOX cells with tyrosine kinase inhibitors as well as microinjection of antibodies directed against p190 but not by non-perturbing antibodies or control buffers. We suggest that activation of the alpha6beta1 integrin signaling regulates the tyrosine phosphorylation state of p190 which in turn connects downstream signaling pathways through Rho family GTPases to actin cytoskeleton in invadopodia, thus promoting membrane-protrusive and degradative activities necessary for cell invasion. PMID- 9417038 TI - A novel integrin-activated pathway forms PKB/Akt-stimulatory phosphatidylinositol 3,4-bisphosphate via phosphatidylinositol 3-phosphate in platelets. AB - The aggregation of human platelets is an important physiological hemostatic event contingent upon receptor-dependent activation of the surface integrin alphaIIbbeta3 and subsequent binding of fibrinogen. Aggregating platelets form phosphatidylinositol 3, 4-bisphosphate (PtdIns(3,4)P2), which has been reported to stimulate in vitro the activity of the proto-oncogenic protein kinase PKB/Akt, as has phosphatidylinositol 3,4,5-trisphosphate (PtdIns(3,4,5)P3). It has been assumed that PtdIns(3,4)P2 is synthesized by either 5-phosphatase-catalyzed hydrolysis of PtdIns(3,4,5)P3 produced by phosphoinositide 3-kinase (PI3K) or phosphorylation by PI3K of PtdIns4P. We investigated the route(s) by which PtdIns(3,4)P2 is formed after directly activating alphaIIbbeta3 with anti-ligand induced binding site Fab fragment and report that aggregation does not lead to the generation of PtdIns(3,4,5)P3, but to transient formation of PtdIns3P and generation of PtdIns(3,4)P2, the latter primarily by PtdIns3P 4-kinase. Both this novel pathway and the activation of PKB/Akt are inhibited by the PI3K inhibitor, wortmannin, and the calpain inhibitor, calpeptin, constituting the first evidence that PtdIns(3,4)P2 can stimulate PKB/Akt in vivo in the absence of PtdIns(3,4,5)P3. Integrin-activated generation of the second messenger PtdIns(3,4)P2 thus depends upon a route distinct from that known to be utilized initially by growth factors. This pathway is of potential general relevance to the function of integrins. PMID- 9417039 TI - Liver microsomal glucose-6-phosphatase is competitively inhibited by the lipid products of phosphatidylinositol 3-kinase. AB - We have studied the effect of various phospholipids on the activity of glucose-6 phosphatase (Glc6Pase) in untreated and detergent-treated rat liver microsomes. Glc6Pase is inhibited in the presence of phosphoinositides in a dose-dependent manner within a range of concentration 0.5-10 microM. The order of efficiency in untreated microsomes is: phosphatidylinositol (PI) 3,4,5P3 > PI3,4P2 = PI4,5P2 > PI3P = PI4P > PI. In contrast, Glc6Pase is not inhibited in the presence of phosphatidylserine, phosphatidylcholine, and phosphatidylethanolamine, diacylglycerol, and inositol 1,4, 5-trisphosphate at concentrations up to 100 microM. The mechanism of Glc6Pase inhibition by PI4,5P2, PI3,4P2, and PI3,4,5P3 is competitive in both untreated and detergent-treated microsomes. In untreated microsomes, the Ki for PI3,4,5P3 (1.7 +/- 0.3 microM, mean +/- S.D. n = 3) is significantly lower (p < 0.01) than that for PI3, 4P2 (5.0 +/- 0.8 microM) and for PI4,5P2 (4.7 +/- 0.7 microM). In detergent-treated microsomes, Glc6Pase is less sensitive to the inhibition and there is no difference anymore among the Ki values for the three compounds: 8.3 +/- 0.8, 11.1 +/- 0.5 and 8.9 +/- 0.4 microM for PI3,4,5P3, PI3,4P2, and PI4,5P2, respectively. This inhibition phenomenon might be of special importance with regards to the insulin's inhibition of hepatic glucose production. PMID- 9417040 TI - Peptide mimics as substrates for the intestinal peptide transporter. AB - 4-Aminophenylacetic acid (4-APAA), a peptide mimic lacking a peptide bond, has been shown to interact with a proton-coupled oligopeptide transporter using a number of different experimental approaches. In addition to inhibiting transport of labeled peptides, these studies show that 4-APAA is itself translocated. 4 APAA transport across the rat intact intestine was stimulated 18-fold by luminal acidification (to pH 6.8) as determined by high performance liquid chromatography (HPLC); in enterocytes isolated from mouse small intestine the intracellular pH was reduced on application of 4-APAA, as shown fluorimetrically with the pH indicator carboxy-SNARF; 4-APAA trans-stimulated radiolabeled peptide transport in brush-border membrane vesicles isolated from rat renal cortex; and in Xenopus oocytes expressing PepT1, 4-APAA produced trans-stimulation of radiolabeled peptide efflux, and as determined by HPLC, was a substrate for translocation by this transporter. These results with 4-APAA show for the first time that the presence of a peptide bond is not a requirement for rapid translocation through the proton-linked oligopeptide transporter (PepT1). Further investigation will be needed to determine the minimal structural requirements for a molecule to be a substrate for this transporter. PMID- 9417041 TI - ARNO is a guanine nucleotide exchange factor for ADP-ribosylation factor 6. AB - ADP-ribosylation factors (ARFs) constitute a family of small monomeric GTPases. ARFs 1 and 3 function in the recruitment of coat proteins to membranes of the Golgi apparatus, whereas ARF6 is localized to the plasma membrane, where it appears to modulate both the assembly of the actin cytoskeleton and endocytosis. Like other GTPases, ARF activation is facilitated by specific guanine nucleotide exchange factors (GEFs). ARNO (ARF nucleotide-binding site opener) is a member of a growing family of ARF-GEFs that share a common, tripartite structure consisting of an N-terminal coiled-coil domain, a central domain with homology to the yeast protein Sec7p, and a C-terminal pleckstrin homology domain. Recently, ARNO and its close homologue cytohesin-1 were found to catalyze in vitro nucleotide exchange on ARF1 and ARF3, respectively, raising the possibility that these GEFs function in the Golgi. However, the actual function of these proteins may be determined in part by their ability to interact with specific ARFs and in part by their subcellular localization. We report here that in vitro ARNO can stimulate nucleotide exchange on both ARF1 and ARF6. Furthermore, based on subcellular fractionation and immunolocalization experiments, we find that ARNO is localized to the plasma membrane in mammalian cells rather than the Golgi. It is therefore likely that ARNO functions in plasma membrane events by modulating the activity of ARF6 in vivo. These findings are consistent with the previous observation that cytohesin-1 regulates the adhesiveness of alphaLbeta2 integrins at the plasma membrane of lymphocytes. PMID- 9417042 TI - Activation of the Jak2-Stat5 signaling pathway in Nb2 lymphoma cells by an anti apoptotic agent, aurintricarboxylic acid. AB - Biological effects of many hormones and cytokines are mediated through receptor associated Jak tyrosine kinases and cytoplasmic Stat transcription factors, including critical physiological processes such as immunity, reproduction, and cell growth and differentiation. Pharmaceuticals that control Jak-Stat pathways are therefore of considerable interest. Here we demonstrate that a single Jak Stat pathway can be activated by aurintricarboxylic acid (ATA), a negatively charged triphenylmethane derivative (475 Da) with anti-apoptotic properties. In prolactin (PRL)-dependent Nb2 lymphocytes, ATA sustained cell growth in the absence of hormone and mimicked rapid PRL-induced tyrosine phosphorylation of Jak2 and activation of Stat5a and Stat5b with tyrosine phosphorylation, heterodimerization, DNA binding, and induction of the Stat5-regulated pim-1 protooncogene. ATA also mimicked PRL activation of serine kinases ERK1 and ERK2. However, unlike PRL, ATA did not regulate Stat1 or Stat3. ATA also did not affect Jak3, which is activated in these cells by interleukin-2 family cytokines. Although the mechanism and specificity by which ATA activates Jak2, Stat5, and ERKs in Nb2 cells are still unclear, the present study demonstrates that certain hormone or cytokine effects on Jak-Stat pathways can be discretely imitated by a low molecular weight, non-peptide pharmaceutical. The results are also consistent with Stat5 involvement in lymphocyte growth and survival. PMID- 9417044 TI - Proximal promoter sequences mediate cell-specific and elevated expression of the favorable prognosis marker TrkA in human neuroblastoma cells. AB - The nerve growth factor receptor, TrkA, has a critical role in the survival, differentiation, and function of neurons in the peripheral and central nervous systems. Recent studies have demonstrated a strong correlation between abundant expression of TrkA and a favorable prognosis of the pediatric tumor, neuroblastoma. This correlation suggests that TrkA may actively promote growth arrest and differentiation of neuroblastoma tumor cells and may be an important therapeutic target in the treatment of this disease. In the present study, we have examined the mechanistic basis for TrkA gene expression in human neuroblastoma cells. Northern blotting and nuclear run-on analyses demonstrated that transcription is a primary determinant of both cell-specific and variable expression of the TrkA gene in neuroblastoma cell lines that express it to different degrees. Cell-specific and variable transcription in neuroblastoma cells was recapitulated by transient transfection of TrkA promoter-luciferase reporter constructs, and regulatory sequences mediating these processes were localized to a 138-base pair region lying just upstream of the transcription initiation region. This neuroblastoma regulatory region formed multiple DNA protein complexes in gel shift assays that were highly enriched in neuroblastoma cells exhibiting abundant TrkA expression. Thus, TrkA-positive neuroblastoma cells are distinguished by differential expression of putative transcription factors that ultimately may serve as targets for up-regulating TrkA expression in tumors with poor prognosis. PMID- 9417043 TI - kappa-Conotoxin PVIIA is a peptide inhibiting the shaker K+ channel. AB - kappa-Conotoxin PVIIA (kappa-PVIIA), a 27-amino acid toxin from Conus purpurascens venom that inhibits the Shaker potassium channel, was chemically synthesized in a biologically active form. The disulfide connectivity of the peptide was determined. kappa-Conotoxin PVIIA has the following structure. This is the first Conus peptide known to target K+ channels. [structure: see text] Although the Shaker K+ channel is sensitive to kappa-PVIIA, the rat brain Kv1.1 subtype is resistant. Chimeras between Shaker and the Kv1.1 K+ channels were constructed and expressed in Xenopus oocytes. Only channels containing the putative pore-forming region between the fifth and sixth transmembrane domains of Shaker retained toxin sensitivity, indicating that the toxin target site is in this region of the channel. Evidence is presented that kappa-PVIIA interacts with the external tetraethyl-ammonium binding site on the Shaker channel. Although both kappa-PVIIA and charybdotoxin inhibit the Shaker channel, they must interact differently. The F425G Shaker mutation increases charybdotoxin affinity by 3 orders of magnitude but abolishes kappa-PVIIA sensitivity. The precursor sequence of kappa-PVIIA was deduced from a cDNA clone, revealing a prepropeptide comprising 72 amino acids. The N-terminal region of the kappa-PVIIA prepropeptide exhibits striking homology to the omega-, muO-, and delta-conotoxins. Thus, at least four pharmacologically distinct superfamilies of Conus peptides belong to the same "O" superfamily, with the omega- and kappa-conotoxins forming one branch, and the delta- and muO-conotoxins forming a second major branch. PMID- 9417045 TI - Direct measurement of the substrate preference of uracil-DNA glycosylase. AB - Site-directed mutants of the herpes simplex virus type 1 uracil-DNA glycosylase lacking catalytic activity have been used to probe the substrate recognition of this highly conserved and ubiquitous class of DNA-repair enzyme utilizing surface plasmon resonance. The residues aspartic acid-88 and histidine-210, implicated in the catalytic mechanism of the enzyme (Savva, R., McAuley-Hecht, K., Brown, T., and Pearl, L. (1995) Nature 373, 487-493; Slupphaug, G., Mol, C. D., Kavli, B., Arvai, A. S., Krokan, H. E. and Tainer, J. A. (1996) Nature 384, 87-92) were separately mutated to asparagine to allow investigations of substrate recognition in the absence of catalysis. The mutants were shown to be correctly folded and to lack catalytic activity. Binding to single- and double-stranded oligonucleotides, with or without uracil, was monitored by real-time biomolecular interaction analysis using surface plasmon resonance. Both mutants exhibited comparable rates of binding and dissociation on the same uracil-containing substrates. Interaction with single-stranded uracil-DNA was found to be stronger than with double stranded uracil-DNA, and the binding to Gua:Ura mismatches was significantly stronger than that to Ade:Ura base pairs suggesting that the stability of the base pair determines the efficiency of interaction. Also, there was negligible interaction between the mutants and single- or double-stranded DNA lacking uracil, or with DNA containing abasic sites. These results suggest that it is uracil in the DNA, rather than DNA itself, that is recognized by the uracil-DNA glycosylases. PMID- 9417046 TI - Heparan sulfate undergoes specific structural changes during the progression from human colon adenoma to carcinoma in vitro. AB - We report a detailed analysis of heparan sulfate (HS) structure using a model of human colon carcinogenesis. Metabolically radiolabeled HS was isolated from adenoma and carcinoma cells. The chain length of HS was the same in both cell populations (Mr 20,000; 45-50 disaccharides), and the chains contained on average of two sulfated domains (S domains), identified by heparinase I scission. This enzyme produced fragments of approximate size 7 kDa, suggesting that the S domains were evenly spaced in the intact HS chain. The degree of polymer sulfation and the patterns of sulfation were strikingly different between the two HS species. When compared with adenoma HS, the iduronic acid 2-O-sulfate content of the carcinoma-derived material was reduced by 33%, and the overall level of N sulfation was reduced by 20%. However, the level of 6-O-sulfation was increased by 24%, and this was almost entirely attributable to an enhanced level of N sulfated glucosamine 6-O-sulfate, a species whose data implied was mainly located in the mixed sequences of alternating N-sulfated and N-acetylated disaccharides. The results indicate that in the transition to malignancy in human colon adenoma cells, the overall molecular organization of HS is preserved, but there are distinct modifications in both the S domains and their flanking mixed domains that may contribute to the aberrant behavior of the cancer cell. PMID- 9417047 TI - Genomic cloning and expression of three murine UDP-galactose: beta-N acetylglucosamine beta1,3-galactosyltransferase genes. AB - Based on the detection of expressed sequence tags that are similar to known galactosyltransferase sequences, we have isolated three novel UDP-galactose:beta N-acetylglucosamine beta1, 3-galactosyltransferase (beta3GalT) genes from a mouse genomic library. The three genes, named beta3GalT-I, -II, and -III, encode type II transmembrane proteins of 326, 422, and 331 amino acids, respectively. The three proteins constitute a distinct subfamily as they do not share any sequence identity with other eucaryotic galactosyltransferases. Also, the entire protein coding region of the three beta3GalT genes was contained in a single exon, which contrasts with the genomic organization of the beta1,4- and alpha1, 3 galactosyltransferase genes. The three beta3GalT genes were mainly expressed in brain tissue. The expression of the full-length murine genes as recombinant baculoviruses in insect cells revealed that the beta3GalT enzymes share the same acceptor specificity for beta-linked GlcNAc, although they differ in their Km for this acceptor and the donor UDP-Gal. The identification of beta3GalT genes emphasizes the structural diversity present in the galactosyltransferase gene family. PMID- 9417048 TI - Specificity of mouse GM2 activator protein and beta-N-acetylhexosaminidases A and B. Similarities and differences with their human counterparts in the catabolism of GM2. AB - Tay-Sachs disease, an inborn lysosomal disease featuring a buildup of GM2 in the brain, is caused by a deficiency of beta-hexosaminidase A (Hex A) or GM2 activator. Of the two human lysosomal Hex isozymes, only Hex A, not Hex B, cleaves GM2 in the presence of GM2 activator. In contrast, mouse Hex B has been reported to be more active than Hex A in cleaving GM2 (Burg, J., Banerjee, A., Conzelmann, E., and Sandhoff, K. (1983) Hoppe Seyler's Z. Physiol. Chem. 364, 821 829). In two independent studies, mice with the targeted disruption of the Hexa gene did not display the severe buildup of brain GM2 or the concomitant abnormal behavioral manifestations seen in human Tay-Sachs patients. The results of these two studies were suggested to be attributed to the reported GM2 degrading activity of mouse Hex B. To clarify the specificity of mouse Hex A and Hex B and to better understand the observed results of the mouse model of Tay-Sachs disease, we have purified mouse liver Hex A and Hex B and also prepared the recombinant mouse GM2 activator. Contrary to the findings of Burg et al., we found that the specificities of mouse Hex A and Hex B toward the catabolism of GM2 were not different from the corresponding human Hex isozymes. Mouse Hex A, but not Hex B, hydrolyzes GM2 in the presence of GM2 activator, whereas GM2 is refractory to mouse Hex B with or without GM2 activator. Importantly, we found that, in contrast to human GM2 activator, mouse GM2 activator could effectively stimulate the hydrolysis of GA2 by mouse Hex A and to a much lesser extent also by Hex B. These results provide clear evidence on the existence of an alternative pathway for GM2 catabolism in mice by converting GM2 to GA2 and subsequently to lactosylceramide. They also provide the explanation for the lack of excessive GM2 accumulation in the Hexa gene-disrupted mice. PMID- 9417049 TI - Induction of c-fos proto-oncogene in mesangial cells by cadmium. AB - Cadmium is mitogenic under some circumstances and has been shown to cause accumulation of transcripts for several proto-oncogenes in a variety of cells, but the mechanism(s) remain to be delineated. Here we show that CdCl2 causes an increase in c-fos mRNA within 30 min of exposure of mesangial cells. At 10 microM Cd2+, this increase persists for at least 8 h in both rat and human cells. The half-life of c-fos mRNA is the same whether it accumulates following 4 h of treatment with Cd2+ or is induced transiently by phorbol ester. Cycloheximide, which stabilizes the transcript, causes a synergistic increase when administered with CdCl2. Nuclear run-on analysis confirms that Cd2+ causes transcriptional activation of the c-fos gene. Calmodulin and Ca2+/calmodulin-dependent kinase, and classical protein kinase C (PKC) isoforms represent two Ca2+-dependent signaling pathways that can lead to induction of c-fos, and Cd2+ has been shown to activate both calmodulin and PKC in vitro, possibly by virtue of the similar ionic radii of Cd2+ and Ca2+. Therefore, we investigated the effect of Cd2+ on these pathways in vivo. 10 microM CdCl2 did not increase total PKC activity or Ca2+/calmodulin-dependent kinase II activity and inhibited the latter at higher concentrations, ruling out either pathway in the Cd2+-dependent induction of c fos. However, Cd2+ did lead to a sustained activation of the Erk family mitogen activated protein kinases (MAPK) that correlated with induction of c-fos. A specific inhibitor of the MAPK kinases, PD98059, partially inhibited the induction of c-fos by Cd2+. We conclude that Cd2+ induces c-fos at least in part by causing a sustained activation of MAPK independent of its ability to activate PKC and calmodulin in vitro. PMID- 9417050 TI - A specific interaction between the cardiac sodium channel and site-3 toxin anthopleurin B. AB - The polypeptide neurotoxin anthopleurin B (ApB) isolated from the venom of the sea anemone Anthopleura xanthogrammica is one of a family of toxins that bind to the extracellular face of voltage-dependent sodium channels and retard channel inactivation. Because most regions of the sodium channel known to contribute to inactivation are located intracellularly or within the membrane bilayer, identification of the toxin/channel binding site is of obvious interest. Recently, mutation of a glutamic acid residue on the extracellular face of the fourth domain of the rat neuronal sodium channel (rBr2a) was shown to disrupt toxin/channel binding (Rogers, J. C., Qu, Y. S., Tanada, T. N., Scheuer, T., and Catterall, W. A. (1996) J. Biol. Chem. 271, 15950-15962). A negative charge at this position is highly conserved between mammalian sodium channel isoforms. We have constructed mutations of the corresponding residue (Asp-1612) in the rat cardiac channel isoform (rH1) and shown that the lowered affinity occurs primarily through an increase in the toxin/channel dissociation rate koff. Further, we have used thermodynamic mutant cycle analysis to demonstrate a specific interaction between this anionic amino acid and Lys-37 of ApB (DeltaDeltaG = 1.5 kcal/mol), a residue that is conserved among many sea anemone toxins. Reversal of the charge at Asp-1612, as in the mutant D1612R, also affects channel inactivation independent of toxin (-14 mV shift in channel availability). Binding of the toxin to Asp-1612 may therefore contribute both to toxin/channel affinity and to transduction of the effects of the toxin on channel kinetics. PMID- 9417051 TI - Identification and molecular characterization of an efflux pump involved in Pseudomonas putida S12 solvent tolerance. AB - Bacteria able to grow in aqueous:organic two-phase systems have evolved resistance mechanisms to the toxic effects of solvents. One such mechanism is the active efflux of solvents from the cell, preserving the integrity of the cell interior. Pseudomonas putida S12 is resistant to a wide variety of normally detrimental solvents due to the action of such an efflux pump. The genes for this solvent efflux pump were cloned from P. putida S12 and their nucleotide sequence determined. The deduced amino acid sequences encoded by the three genes involved show a striking resemblance to proteins known to be involved in proton-dependent multidrug efflux systems. Transfer of the genes for the solvent efflux pump to solvent-sensitive P. putida strains results in the acquisition of solvent resistance. This opens up the possibilities of using the solvent efflux system to construct bacterial strains capable of performing biocatalytic transformations of insoluble substrates in two-phase aqueous:organic medium. PMID- 9417052 TI - Intermolecular NH2-/carboxyl-terminal interactions in androgen receptor dimerization revealed by mutations that cause androgen insensitivity. AB - Structural alignment of the human androgen receptor dimer was investigated by introducing steroid binding domain mutations that cause partial or complete androgen insensitivity into fusion proteins containing the full-length androgen receptor or the steroid binding domain. Most of the mutants had unchanged apparent equilibrium androgen binding affinity and increased dissociation rates of [3H]methyltrienolone and required increased dihydrotestosterone concentrations for transcriptional activation. In a 2-hybrid protein interaction assay in mammalian cells, the steroid binding domain interacts with an NH2-terminal-DNA binding domain fragment and with the full-length androgen receptor at physiological androgen concentrations in a dose-dependent manner. However, mutations at Val-889 and Arg-752 disrupt the NH2-/carboxyl-terminal interaction when introduced into the steroid binding domain fragment but not when present in the full-length androgen receptor. The N-C bimolecular interaction reduces the dissociation rate of bound androgen and slows the degradation rate of the carboxyl-terminal steroid binding domain fragment. The results suggest that steroid binding domain residues Val-889 and Arg-752 are critical to the NH2 /carboxyl-terminal interaction and that an intermolecular N-C interaction occurs during receptor dimerization that results in an antiparallel arrangement of androgen receptor monomers. PMID- 9417053 TI - Cytidylyltransferase-binding protein is identical to transcytosis-associated protein (TAP/p115) and enhances the lipid activation of cytidylyltransferase. AB - We previously identified a protein from rat liver that binds CTP:phosphocholine cytidylyltransferase (CT). We have now purified this protein (cytidylyltransferase-binding protein (CTBP)) from rat liver. The purification involved precipitation at pH 5 and extraction of the precipitate with buffer, followed by sequential chromatography on DEAE-Sepharose and butyl-agarose. Final purification was accomplished by either preparative electrophoresis or hydroxylapatite chromatography. Amino acid sequences from six peptides derived from pure CTBP matched sequences in transcytosis-associated protein (TAP) with 98% identity. Thus, CTBP was positively identified to be TAP. Purified CTBP increased the activity of purified CT measured with phosphatidylcholine (PC)/oleic acid. In the absence of PC/oleic acid, CTBP did not stimulate CT activity. Dilution of CT to reduce the Triton X-100 concentration produced a loss of CT activity. The lost activity was recovered by the addition of CTBP plus PC/oleic acid to the assay, but not by the addition of either PC/oleic acid or CTBP alone. Removal of CTBP from purified preparations by immunoprecipitation with CTBP antibodies eliminated the activation of CT. Both CT and CTBP were shown to bind to PC/oleic acid liposomes. The formation of complexes between CT and CTBP in the absence of PC/oleic acid liposomes could not be demonstrated. These results suggest that CTBP functions to modify the interaction of CT with PC/oleic acid liposomes, resulting in an increase in the catalytic activity perhaps by the formation of a ternary complex between CT, CTBP, and lipid. Overall, these results suggest that CTBP (TAP) may function to coordinate the biosynthesis of phosphatidylcholine with vesicle transport. PMID- 9417054 TI - A novel ets-related transcription factor, ERT/ESX/ESE-1, regulates expression of the transforming growth factor-beta type II receptor. AB - A 2.5-kilobase cDNA clone that encodes a 371-amino acid novel transcription factor was isolated from a human placenta cDNA library using a yeast one-hybrid system. The novel ets-related transcription factor (ERT) showed a homology with the ETS DNA-binding domain. Using constructs of the transforming growth factor beta (TGF-beta) type II receptor (RII) promoter linked to the luciferase gene, we have demonstrated that ERT activates transcription of the TGF-beta RII gene through the 5'-TTTCCTGTTTCC-3' response element spanning nucleotides +13 to +24 and multiple additional ETS binding sites between -1816 and -82 of the TGF-beta RII promoter. A specific interaction between ERT and the ETS binding sites was also demonstrated using an electrophoretic mobility shift assay. Deletion mapping of ERT protein suggests that the transactivation domain resides in the amino terminus while the DNA-binding domain is localized to the carboxyl-terminal region. Our results suggest that ERT might be a major transcription factor involved in the transcriptional regulation of the TGF-beta RII gene. PMID- 9417055 TI - Cyclic GMP-dependent and -independent effects on the synthesis of the calcium messengers cyclic ADP-ribose and nicotinic acid adenine dinucleotide phosphate. AB - Cyclic ADP-ribose (cADPR) and nicotinic acid adenine dinucleotide phosphate (NAADP) have been shown to mobilize intracellular Ca2+ stores by totally independent mechanisms, which are pharmacologically distinct from that activated by inositol trisphosphate. Although cADPR and NAADP are structurally and functionally different, they can be synthesized by a single enzyme having ADP ribosyl cyclase activity. In this study, three different assays were used to measure the metabolism of cADPR in sea urchin egg homogenates including a radioimmunoassay, a Ca2+ release assay, and a thin layer chromatographic assay. Soluble and membrane-bound ADP-ribosyl cyclases were identified and both cyclized NAD to produce cADPR. The soluble cyclase was half-maximally stimulated by 5.3 microM cGMP, but not by cAMP, while the membrane-bound form was independent of cGMP. The two forms of the cyclase were also different in the pH dependence of utilizing nicotinamide guanine dinucleotide (NGD), a guanine analog of NAD, as substrate, indicating they are two separate enzymes. The stimulatory effect of cGMP required ATP or ATPgammaS (adenosine 5'-O-(3-thiotriphosphate)) and a cGMP dependent kinase activity was shown to be present in the soluble fraction. The degradation of cADPR to ADP-ribose was catalyzed by cADPR hydrolase, which was found to be predominantly associated with membranes. Similar to the membrane bound cyclase, the cADPR hydrolase activity was also independent of cGMP. Both the soluble and membrane fractions also catalyzed the synthesis of NAADP through exchanging the nicotinamide group of NADP with nicotinic acid (NA). The base exchange activity was independent of cGMP and the half-maximal concentrations of NADP and NA needed were about 0.2 mM and 10 mM, respectively. The exchange reaction showed a preference for acidic pH, contrasting with the neutral pH optimum of the cyclase activities. The complex metabolic pathways characterized in this study indicate that there may be a multitude of regulatory mechanisms for controlling the endogenous concentrations of cADPR and NAADP. PMID- 9417056 TI - Overexpression of cholesterol 7alpha-hydroxylase (CYP7A) in mice lacking the low density lipoprotein (LDL) receptor gene. LDL transport and plasma LDL concentrations are reduced. AB - This study was undertaken to determine the effect of transient overexpression of hepatic cholesterol 7alpha-hydroxylase on low density lipoprotein (LDL) cholesterol transport in mice lacking LDL receptors (LDL receptor-/-). Primary overexpression of hepatic 7alpha-hydroxylase in LDL receptor-/- mice was accompanied by a dose-dependent decrease in the rate of LDL cholesterol appearance in plasma (whole body LDL cholesterol transport) and a corresponding reduction in circulating LDL cholesterol levels. The increase in hepatic 7alpha hydroxylase activity necessary to achieve a 50% reduction in plasma LDL cholesterol concentrations was approximately 10-fold. In comparison, cholestyramine increased hepatic 7alpha-hydroxylase activity approximately 3-fold and reduced plasma LDL cholesterol concentrations by 17%. This study demonstrates that augmentation of hepatic 7alpha-hydroxylase expression is an effective strategy for lowering plasma LDL concentrations even in animals with a genetic absence of LDL receptors. PMID- 9417057 TI - Receptor-activated Ca2+ influx via human Trp3 stably expressed in human embryonic kidney (HEK)293 cells. Evidence for a non-capacitative Ca2+ entry. AB - Ca2+ release from its internal stores as a result of activation of phospholipase C is accompanied by Ca2+ influx from the extracellular space. Ca2+ influx channels may be formed of proteins homologous to Drosophila Trp. At least six non allelic Trp genes are present in the mouse genome. Full-length human, bovine, mouse, and rat cDNAs for Trp1, 3, 4, 6 have been cloned. Expression of these genes in various mammalian cells has provided evidence that Trp proteins form plasma membrane Ca2+-permeant channels that can be activated by an agonist that activates phospholipase C, by inositol 1,4, 5-trisphosphate, and/or store depletion. We have stably expressed human Trp3 (hTrp3) in human embryonic kidney (HEK)293 cells. Measurement of intracellular Ca2+ concentrations in Fura2-loaded cells showed that cell lines expressing hTrp3 have significantly higher basal and agonist-stimulated influxes of Ca2+, Mn2+, Ba2+, and Sr2+ than control cells. The increase in Ca2+ entry attributable to the expression of hTrp3 obtained upon store depletion by thapsigargin was much lower than that obtained by stimulation with agonists acting via a Gq-coupled receptor. Addition of agonists to thapsigargin-treated Trp3 cells resulted in a further increase in the entry of divalent cations. The increased cation entry in Trp3 cells was blocked by high concentrations of SKF 96365, verapamil, La3+, Ni2+, and Gd3+. The Trp3-mediated Ca2+ influx activated by agonists was inhibited by a phospholipase C inhibitor, U73122. We propose that expression of hTrp3 in these cells forms a non-selective cation channel that opens after the activation of phospholipase C but not after store depletion. In addition, a subpopulation of the expressed hTrp3 may form heteromultimeric channels with endogenous proteins that are sensitive to store depletion. PMID- 9417058 TI - Intact vitronectin induces matrix metalloproteinase-2 and tissue inhibitor of metalloproteinases-2 expression and enhanced cellular invasion by melanoma cells. AB - The initial site of melanoma cell metastasis is frequently the regional lymph nodes, and the appearance of lymph node metastasis correlates with poor prognosis. Lymph node adhesion is mediated by an interaction between the tumor cell integrin alphavbeta3 and lymph node vitronectin. In this study, we explored the relationship between adhesion and proteolysis by examining the direct effect of vitronectin receptor ligation on matrix metalloproteinase-2 (MMP-2) production by B16F1 and B16F10 melanoma cells. We report a dose-dependent increase in secretion of both MMP-2 and tissue inhibitor of metalloproteinases-2 (TIMP-2) in response to vitronectin. Cellular invasiveness was also enhanced by vitronectin, as shown by the increased ability of vitronectin-treated cells to invade a synthetic basement membrane (Matrigel). Both the vitronectin-induced MMP-2 production and vitronectin-enhanced invasion were blocked by the peptide ligand Arg-Gly-Asp-Ser (RGDS). Furthermore, neither plasmin-degraded vitronectin nor the peptide ligand RGDS stimulated MMP-2 secretion or invasiveness, indicating that a multivalent ligand-receptor interaction rather than simple receptor occupancy was required for MMP-2 induction. MMP-2 and MMP-2/TIMP-2 interaction with the plasma membrane of melanoma cells resulted in enhanced catalytic activity against 14C labeled gelatin, suggesting that membrane association may function in posttranslational regulation of MMP-2 activity. This is supported by data showing increased cellular invasion by cells containing membrane-bound MMP-2. Binding of proMMP-2 and proMMP-2/TIMP-2 to melanoma cells was not inhibited by RGDS, and melanoma cell adhesion to vitronectin was unaffected by pro- or active MMP-2, indicating that MMP-2 did not interact with the murine vitronectin receptor. Together, these data provide evidence for a functional link between adhesion and proteolysis and suggest a potential mechanism whereby adhesion of an invasive cell to the extracellular matrix regulates subsequent invasive behavior. PMID- 9417059 TI - Mutational analysis of the nucleotide binding sites of the yeast vacuolar proton translocating ATPase. AB - To further define the structure of the nucleotide binding sites on the vacuolar proton-translocating ATPase (V-ATPase), the role of aromatic residues at the catalytic sites was probed using site-directed mutagenesis of the VMA1 gene that encodes the A subunit in yeast. Substitutions were made at three positions (Phe452, Tyr532, and Phe538) that correspond to residues observed in the crystal structure of the homologous beta subunit of the bovine mitochondrial F-ATPase to be in proximity to the adenine ring of bound ATP. Although conservative substitutions at these positions had relatively little effect on V-ATPase activity, replacement with nonaromatic residues (such as alanine or serine) caused either a complete loss of activity (F452A) or a decrease in the affinity for ATP (Y532S and F538A). The F452A mutation also appeared to reduce stability of the V-ATPase complex. These results suggest that aromatic or hydrophobic residues at these positions are essential to maintain activity and/or high affinity binding to the catalytic sites of the V-ATPase. Site-directed mutations were also made at residues (Phe479 and Arg483) that are postulated to be contributed by the A subunit to the noncatalytic nucleotide binding sites. Generally, substitutions at these positions led to decreases in activity ranging from 30 to 70% relative to wild type as well as modest decreases in Km for ATP. Interestingly, the R483E and R483Q mutants showed a time-dependent increase in ATPase activity following addition of ATP, suggesting that events at the noncatalytic sites may modulate the catalytic activity of the enzyme. PMID- 9417060 TI - A novel, secreted form of human ADAM 12 (meltrin alpha) provokes myogenesis in vivo. AB - The ADAM (A Disintegrin And Metalloprotease) family of cell-surface proteins may have an important role in cellular interactions and in modulating cellular responses. In this report we describe a novel, secreted form of human ADAM 12 (meltrin alpha), designated ADAM 12-S (S for short), and a larger, membrane-bound form designated ADAM 12-L (L for long form). These two forms arise by alternative splicing of a single gene located on chromosome 10q26. Northern blotting demonstrated that mRNAs of both forms are abundant in human term placenta and are also present in some tumor cell lines. The ADAM 12-L transcript can also be detected in normal human adult skeletal, cardiac, and smooth muscle. Human A204 embryonal rhabdomyosarcoma cells that do not differentiate into muscle cells and do not express any form of ADAM 12 were stably transfected with an ADAM 12-S minigene encoding the disintegrin domain, the cysteine-rich domain, and the unique 34 amino acid carboxyl terminus. Nude mouse tumors derived from these transfected cells contained ectopic muscle cells of apparent mouse origin as shown by species-specific markers. These results may have potential applications in the development of muscle-directed gene and cell therapies. PMID- 9417061 TI - Control of the structural stability of the tubulin dimer by one high affinity bound magnesium ion at nucleotide N-site. AB - Tubulin liganded with GTP at the N-site in the alpha-subunit and with GDP at the E-site in the beta-subunit (GDP-tubulin) reversibly binds one high affinity Mg2+ cation (Kb = 1.1 x 10(7) M-1), whereas tubulin liganded with GTP at both subunits (GTP-tubulin) binds one more high affinity Mg2+. The two cation binding loci are identified as nucleotide sites N and E, respectively. Mg2+ at the N-site controls the stability and structure of the alphabeta-tubulin dimer. Mg2+ dissociation is followed by the slow release of bound nucleotide and functional inactivation. Mg2+ bound to the N-site significantly increases the thermal stability of the GDP tubulin dimer (by 10 degrees C and approximately 50 kcal mol-1 of experimental enthalpy change). However, the thermal stability of Mg2+-liganded GDP- and GTP tubulin is the same. Mg2+ binding to the N-site is linked to the alphabeta-dimer formation. The binding of Mg2+ to the alpha-subunit communicates a marked enhancement of fluorescence to a colchicine analogue bound to the beta-subunit. Colchicine, in turn, thermally stabilizes Mg2+-depleted tubulin. The tubulin properties described would be simply explained if the N-site and the colchicine site are at the alpha-beta dimerization interface. It follows that the E-site would be at the beta-end of the tubulin dimer, consistent with the known functional role of the E nucleotide gamma-phosphate and coordinated cation controlling microtubule stability. PMID- 9417062 TI - The dynamic properties of the M121H azurin metal site as studied by NMR of the paramagnetic Cu(II) and Co(II) metalloderivatives. AB - The M121H azurin mutant in solution presents various species in equilibrium that can be detected and studied by 1H NMR of the Cu(II) and Co(II) paramagnetic metalloderivatives. In both cases up to three species are observed in slow exchange, the proportions of which are different for the two metalloderivatives. Above pH 5 the major species displays a tetrahedral coordination in which the His121 can be observed as a coordinated residue. Its metal site corresponds to a new type of site that is defined as a type 1.5 site. The second and third species resemble the wild type (type 1) azurin and, above pH 4.5, they are present only at a low concentration. At low pH a protonation process increases the proportion of both type 1 species at the expense of the type 1.5 species. This process, characterized by a pKa = 4.3, is assigned to the protonation of His121. At high pH the NMR spectrum of the Co(II)-M121H azurin experiences an additional transition, which is not observed in the case of the Cu(II) protein. The dynamic properties of the M121H metal site appear to be related to changes in the coordination geometry and the strength of the axial interaction between the Ndelta1 (His121) and the metal. PMID- 9417063 TI - Basolateral sorting of the cation-dependent mannose 6-phosphate receptor in Madin Darby canine kidney cells. Identification of a basolateral determinant unrelated to clathrin-coated pit localization signals. AB - In polarized Madin-Darby canine kidney (MDCK) cells, sorting of membrane proteins in the trans-Golgi network for basolateral delivery depends on the presence of cytoplasmic determinants that are related or unrelated to clathrin-coated pit localization signals. Whether these signals mediate basolateral protein sorting through common or distinct pathways is unknown. The cytoplasmic domain of the cation-dependent mannose 6-phosphate receptor (CD-MPR) contains clathrin-coated pit localization signals that are necessary for endocytosis and lysosomal enzyme targeting. In this study, we have addressed the function of these signals in polarized sorting of the CD-MPR. A chimeric protein, made of the luminal domain of the influenza virus hemagglutinin fused to the transmembrane and cytoplasmic domains of the CD-MPR was stably expressed in MDCK cells. This chimera (HCD) is able to interact with the AP-1 Golgi-specific assembly proteins and is detected on the basolateral plasma membrane of MDCK cells where it is endocytosed. Deletion analysis and site-directed mutagenesis of the cytoplasmic domain of the CD-MPR indicate that HCD chimeras devoid of clathrin-coated pit localization signals are still transported to the basolateral membrane where they accumulate. A HCD chimera containing only the transmembrane domain and the 12 membrane proximal amino acids of the CD-MPR cytoplasmic tail is also found on the basolateral membrane but is unable to interact with the AP-1 assembly proteins. However, the overexpression of this mutant results in partial apical delivery. It is concluded, therefore, that the basolateral transport of this chimera requires a saturable sorting machinery distinct from AP-1. PMID- 9417064 TI - Cell cycle regulation of histone H4 gene transcription requires the oncogenic factor IRF-2. AB - Histone genes display a peak in transcription in early S phase and are ideal models for cell cycle-regulated gene expression. We have previously shown that the transcription factor interferon regulatory factor 2 (IRF-2) can activate histone H4 gene expression. In this report we establish that a mouse histone H4 gene and its human homolog lose stringent cell cycle control in synchronized embryonic fibroblasts in which IRF-2 has been ablated. We also show that there are reduced mRNA levels of this endogenous mouse histone H4 gene in the IRF-2(-/ ) cells. Strikingly, the overall mRNA level and cell cycle regulation of histone H4 transcription are restored when IRF-2 is reintroduced to these cells. IRF-2 is a negative regulator of the interferon response and has oncogenic potential, but little is known of the mechanism of these activities. Our results suggest that IRF-2 is an active player in E2F-independent cell cycle-regulated gene expression at the G1/S phase transition. IRF-2 was previously considered a passive antagonist to the tumor suppressor IRF-1 but can now join other oncogenic factors such as c-Myb and E2F1 that are predicted to mediate their transforming capabilities by actively regulating genes necessary for cell cycle progression. PMID- 9417065 TI - Constitutive activation of phosphatidylinositol 3-kinase by a naturally occurring mutant epidermal growth factor receptor. AB - The most frequently found alteration of the epidermal growth factor receptor (EGFR) in human tumors is a deletion of exons 2-7. This receptor, termed EGFRvIII, can transform NIH 3T3 cells, and the frequent expression of this variant implies that it confers a selective advantage upon tumor cells in vivo. Although EGFRvIII is a constitutively activated tyrosine kinase, there is no increase in Ras.GTP levels and low levels of mitogen-activated protein kinase activity in NIH 3T3 cells expressing this variant. We investigated whether phosphatidylinositol (PI) 3-kinase was an effector in transformation by the EGFRvIII. High levels of PI 3-kinase activity were constitutively present in EGFRvIII-transformed cells and were dependent upon the kinase activity of the receptor. While mitogen-activated protein kinase activity was quickly down regulated to basal levels after 12 h of continuous EGFR activation, there was a 3 fold increase in PI 3-kinase activity in cells expressing normal EGFR and an 8 fold increase in cells expressing EGFRvIII after 48 h. This increased activity may reflect enhanced binding to EGFRvIII and the presence of novel PI 3-kinase isoforms. Treatment with the PI 3-kinase inhibitors wortmannin and LY294002 blocked both anchorage-independent growth and growth in low serum media and also resulted in morphological reversion of EGFRvIII-transformed cells. These results support an essential role for PI 3-kinase in transformation by this EGFR variant. PMID- 9417066 TI - Multiple splice variants of the human calcium-independent phospholipase A2 and their effect on enzyme activity. AB - Recently, the cloning of a novel Ca2+-independent phospholipase A2 (iPLA2) from Chinese hamster ovary cells as well as from mouse and rat sources containing a C terminal lipase motif and eight N-terminal ankyrin repeats has been described. In this report we describe the cloning of the human iPLA2 cDNA and its expression in B-cells and show that the iPLA2 gene undergoes extensive alternative splicing generating multiple isoforms that contribute to a novel mechanism to control iPLA2 activity. The full-length cDNA clone encodes a 806-amino acid protein with a calculated molecular mass of 88 kDa. The protein contains a lipase motif, GXSXG, and ankyrin repeats, as described for the hamster and rodent forms of the enzyme but has an additional 54-amino acid proline-rich insertion in the last of the eight ankyrin repeats (residues 395-449). Furthermore, at least three additional isoforms most likely due to alternative splicing were identified. One that is present as a partial cDNA in the expressed sequence tag data base is similar to iPLA2 but terminates just after the lipase active site, and two other isoforms contain only the iPLA2 ankyrin repeat sequence (ankyrin-iPLA2-1 and -2). Ankyrin repeats are involved in protein-protein interactions and because the purified iPLA2 enzyme exists as a multimeric complex of 270-350 kDa, the expression of just the ankyrin-iPLA2 sequence suggested that these may also interact with the iPLA2 oligomeric complexes and perhaps modulate PLA2 activity. Transfection of the human iPLA2 cDNA into COS cells resulted in a substantial increase in calcium-independent PLA2 activity in cell lysate. No activity above background was observed following ankyrin-iPLA2-1 cDNA transfection. However, co transfection of the ankyrin-iPLA2-1 and the iPLA2 cDNAs resulted in a 2-fold reduction in activity compared with iPLA2 alone. A similar co-transfection of ankyrin-iPLA2-1 cDNA with the cPLA2 cDNA had no effect on PLA2 activity. These results suggest that the ankyrin-iPLA2 sequence can function as a negative regulator of iPLA2 activity and that the alternative splicing of the iPLA2 gene can have a direct effect on the attenuation of enzyme activity. PMID- 9417067 TI - Identification of novel phosphorylation sites in hormone-sensitive lipase that are phosphorylated in response to isoproterenol and govern activation properties in vitro. AB - Hormone-sensitive lipase (HSL) is the rate-limiting enzyme in lipolysis. Stimulation of rat adipocytes with isoproterenol results in phosphorylation of HSL and a 50-fold increase in the rate of lipolysis. In this study, we used site directed mutagenesis and two-dimensional phosphopeptide mapping to show that phosphorylation sites other than the previously identified Ser-563 are phosphorylated in HSL in response to isoproterenol stimulation of 32P-labeled rat adipocytes. Phosphorylation of HSL in adipocytes in response to isoproterenol and in vitro phosphorylation of HSL containing Ser --> Ala mutations in residues 563 and 565 (S563A, S565A) with protein kinase A (PKA), followed by tryptic phosphopeptide mapping resulted in two tryptic phosphopeptides. These tryptic phosphopeptides co-migrated with the phosphopeptides released by the same treatment of F654HPRRSSQGVLHMPLYSSPIVK675 phosphorylated with PKA. Analysis of the phosphorylation site mutants, S659A, S660A, and S659A,S660A disclosed that mutagenesis of both Ser-659 and Ser-660 was necessary to abolish the activation of HSL toward a triolein substrate after phosphorylation with PKA. Mutation of Ser-563 to alanine did not cause significant change of activation compared with wild-type HSL. Hence, our results demonstrate that in addition to the previously identified Ser-563, two other PKA phosphorylation sites, Ser-659 and Ser-660, are present in HSL and, furthermore, that Ser-659 and Ser-660 are the major activity controlling sites in vitro. PMID- 9417068 TI - Blood coagulation factor IX residues Glu78 and Arg94 provide a link between both epidermal growth factor-like domains that is crucial in the interaction with factor VIII light chain. AB - Recently, we established that mutations at calcium-binding sites within the first epidermal growth factor (EGF)-like domain of activated factor IX affect its interaction with factor VIIIa (Lenting, P. J., Christophe, O. D., ter Maat, H., Rees, D. J. G., and Mertens, K. (1996) J. Biol. Chem. 271, 25332-25337). In the present study, we have investigated the functional role of residue Glu78, which is not involved in calcium binding. Glu78 is also located in the first EGF-like domain and, when mutated to Lys, is associated with severe hemophilia B. Because Glu78 is conserved in related vitamin K-dependent proteins, it is difficult to understand how a mutation at this position is associated with factor IX-specific function. In this study, we addressed the hypothesis that Glu78 exerts its biological activity by interacting with another residue. One candidate was found to be the second EGF-like domain residue, Arg94, which is also associated with severe hemophilia B when mutated. We constructed a series of mutants that included mutations at position 78 alone (Glu78 to Lys/Glu78 to Asp) or at both positions 78 and 94 (Glu78 to Lys and Arg94 to Asp). The functional parameters of immunopurified and activated mutants were compared with normal activated factor IX. Mutants were indistinguishable from normal factor IXa in cleaving the synthetic substrate CH3SO2-Leu-Gly-Arg-p-nitroanilide or activating factor X in the absence of factor VIIIa. In contrast, in the presence of factor VIIIa, factor IXa Glu78 to Asp and factor IXa Glu78 to Lys/Arg94 to Asp were stimulated to the same extent as normal factor IXa, whereas factor IXa Glu78 to Lys was markedly less stimulated (140-fold versus 2,000-fold). This suggests that residues 78 and 94 should carry an opposite charge for a normal interaction of factor IXa to factor VIIIa. This hypothesis was confirmed in inhibition studies employing synthetic peptides comprising the factor IXa-binding motifs of factor VIII heavy (Ser558-Gln565) or light chain (Glu1811-Lys1818) and in direct binding studies. We propose that residues 78 and 94 link both EGF-like domains and thereby maintain the integrity of the factor VIII light chain binding site. PMID- 9417070 TI - Conserved residues and motifs in the NixA protein of Helicobacter pylori are critical for the high affinity transport of nickel ions. AB - NixA, the high affinity nickel transport protein of Helicobacter pylori, imports Ni2+ ions across the cytoplasmic membrane for insertion into the active site of the urease metalloenzyme, which is essential for colonization of the gastric mucosa. Twelve conserved aspartate (aspartates 47, 49, 55, 194, 231, and 234), glutamate (glutamates 106, 198, and 274), and histidine (histidines 44, 50, and 79) residues were identified by alignment of NixA with homologous transporters. Polymerase chain reaction-generated site-directed mutants of these residues were expressed in E. coli along with the H. pylori urease gene cluster. Mutations in residues within the predicted periplasmic domains of NixA maintained near wild type levels of Ni2+ uptake and urease activity, as did control mutations of conserved positively charged residues (lysines 140 and 268; arginines 162 and 167). Mutations in highly conserved motifs in predicted helices II and III of NixA abolished Ni2+ uptake and urease activity. Mutations in helices V and VI and the cytoplasmic domains decreased Ni2+ transport rates by >/=90%. Reduction in rates of Ni2+ transport correlated with reduction in urease activities (r = 0.77). Ni2+ transport was inhibited in the presence of Co2+, Cu2+, and Zn2+, indicating that these ions may also be bound or transported by NixA. We conclude that conserved Asp, Glu, and His residues in the transmembrane domains of NixA are critical for the transport of the divalent cations Ni2+, Co2+, Cu2+, and Zn2+ into the cytoplasm of H. pylori. PMID- 9417069 TI - Carbon flux via the pentose phosphate pathway regulates the hepatic expression of the glucose-6-phosphatase and phosphoenolpyruvate carboxykinase genes in conscious rats. AB - Hepatic gene expression of P-enolpyruvate carboxykinase (PEPCK) and glucose-6 phosphatase (Glc-6-Pase) is regulated in response to changes in the availability of substrates, in particular glucose (Glc; Massillon, D., Barzilai, N., Chen, W., Hu, M., and Rossetti, L. (1996) J. Biol. Chem. 271, 9871-9874). We investigated the mechanism(s) in conscious rats. Hyperglycemia per se caused a rapid and marked increase in Glc-6-Pase mRNA abundance and protein levels. By contrast, hyperglycemia decreased the abundance of PEPCK mRNA. Importantly, inhibition of glucokinase activity by glucosamine infusion blunted both the stimulation of Glc 6-Pase and the inhibition of PEPCK gene expression by Glc, suggesting that an intrahepatic signal (metabolite) generated by the metabolism of glucose at or beyond Glc-6-P was responsible for the regulatory effect of Glc. The effect of Glc on the L-type pyruvate kinase gene is mediated by xylulose-5-P (Doiron, B., Cuif, M., Chen, R., and Kahn, A. (1996) J. Biol. Chem. 271, 5321-5324). Thus, we next investigated whether an isolated increase in the hepatic concentration of this metabolite can also reproduce the effects of Glc on Glc-6-Pase and PEPCK gene expression in vivo. Xylitol, which is directly converted to xylulose-5-P in the liver, was infused to raise the hepatic concentration of xylulose-5-P by approximately 3-fold. Xylitol infusion did not alter the levels of Glc-6-P and of fructose-2,6-biphosphate. However, it replicated the effects of hyperglycemia on Glc-6-Pase and PEPCK gene expression and resulted in a 75% increase in the in vivo flux through Glc-6-Pase (total glucose output). PMID- 9417071 TI - Sphingosine inhibits voltage-operated calcium channels in GH4C1 cells. AB - In the present study we investigated the mechanism of inhibitory action of sphingosine (SP) on voltage-activated calcium channels (VOCCs) in pituitary GH4C1 cells. Using the patch-clamp technique in the whole-cell mode, we show that SP inhibits Ba2+ currents (IBa) when 0.1 mM BAPTA is included in the patch pipette. However, when the BAPTA concentration was raised to 1-10 mM, SP was without a significant effect. The effect of SP was apparently not mediated via a kinase, as it was not inhibited by staurosporine. By using the double-pulse protocol (to release possible functional inhibition of the VOCCs by G proteins), we observed that G proteins apparently evoked very little functional inhibition of the VOCCs. Furthermore, including GDPbetaS (guanyl-5'-yl thiophosphate) in the patch pipette did not alter the inhibitory effect of SP on the Ba2+ current, suggesting that SP did not modulate the VOCCs via a G protein-dependent pathway. Single-channel experiments with SP in the pipette, and experiments with excised outside-out patches, suggested that SP directly inhibited VOCCs. The main mechanism of action was a dose-dependent prolongation of the closed time of the channels. The results thus show that SP is a potent inhibitor of VOCCs in GH4C1 cells, and that calcium may be a cofactor in this inhibition. PMID- 9417072 TI - Heparinase I from Flavobacterium heparinum. Role of positive charge in enzymatic activity. AB - Heparinases are bacterial enzymes that are powerful tools to study the physiological roles of heparin-like complex polysaccharides. In addition, heparinases have significant therapeutic applications. We had proposed earlier that cysteine 135 and histidine 203 together form the catalytic domain in heparinase I. We had also identified a heparin binding domain in heparinase I containing two positively charged clusters HB-1 and HB-2 in a primary heparin binding site and other positively charged residues in the vicinity of cysteine 135. In this study, through systematic site-directed mutagenesis studies, we show that the alteration of the positive charge of the HB-1 region has a pronounced effect on heparinase I activity. More specifically, site-directed mutagenesis of K199A (contained in HB-1) results in a 15-fold reduction in catalytic activity, whereas a K198A mutation (also in HB-1) results in only a 2- to 3-fold reduction in heparinase I activity. A K132A mutation, in close proximity to cysteine 135, also resulted in reduced (8-fold) activity. Heparin affinity chromatography experiments indicated moderately lowered binding affinities for the K132A, K198A, and the K199A mutant enzymes. The above results, taken together with our previous observations, lead us to propose that the positively charged heparin binding domain provides the necessary microenvironment for the catalytic domain of heparinase I. The dominant effect of lysine 199 suggests an additional, more direct, role in catalysis for this residue. PMID- 9417073 TI - Identification of functionally important residues of human thrombopoietin. AB - Thrombopoietin (TPO) is a megakaryocyte growth and differentiation factor. It consists of a characteristic two domain structure. The amino-terminal domain of TPO has a sequence homology with erythropoietin and is required for the binding and activation of its receptor c-Mpl. To determine the functionally important regions interacting with its receptor, a series of site-directed mutants of TPO were constructed based on a three-dimensional model of the amino-terminal domain. Two strategies of mutagenesis were employed: 1) nonnative N-linked glycosylation scan of 12 residues predicted to be on the surface, and 2) alanine replacement scan of mostly charged 44 amino acid residues. Each TPO mutein was transiently expressed in COS7 cells, and the specific bioactivity of the TPO protein secreted into the culture medium was measured using a recombinant BaF3 cell line expressing human c-Mpl. Four alanine substitutions at Arg10, Pro42, Glu50, and Lys138 nearly or completely abolished the activity, whereas the mutation at Arg14 slightly decreased the activity, suggesting that these residues are functionally important in interacting with its receptor. These residues mapped to helix A, loop AB, and helix D. Sequence comparison between human TPO and other mammalian TPO showed that the identified residues are completely conserved among the species. However, unlike the recent report on the mutational analysis of TPO, alanine substitutions at Lys52, Lys59, Arg136, and Arg140 did not affect the TPO activity significantly in our system. The identified receptor binding regions of TPO are analogous to those of human growth hormone and erythropoietin. Based on the similarity of these three cytokines, we propose that Lys138 of helix D and Pro42 and Glu50 of loop AB may constitute one binding region, whereas Arg10 and Lys14 of helix A may constitute the other binding region to dimerize the receptors. PMID- 9417074 TI - Directly energized uptake of beta-estradiol 17-(beta-D-glucuronide) in plant vacuoles is strongly stimulated by glutathione conjugates. AB - A directly energized vacuolar pump for glutathione (GS) conjugates has been described for several plant species. Since glucuronate conjugates also occur in plants, we addressed the question whether plant vacuoles take up the abiotic glucuronate conjugate estradiol 17-(beta-glucuronide) (E217G) via a GS conjugate pump, which in some cases has been reported to accept various organic anions as substrates, or via a distinct glucuronate transporter. Uptake studies into vacuoles from rye and barley were performed with E217G and metolachlor-GS (MOC GS), a substrate of the GS conjugate ATPase, to compare glucuronate conjugate transport into vacuoles containing endogenous flavone glucuronides with those lacking specific glucuronate conjugates, respectively. Our results indicate that E217G and MOC-GS are taken up into vacuoles of both plants via a directly energized mechanism since transport was (i) strictly ATP-dependent; (ii) inhibited by vanadate but not by bafilomycin A1, azide, verapamil, nor by dissipation of the vacuolar DeltapH or DeltaPsi; (iii) E217G uptake into rye vacuoles was partially driven by other nucleotides in the following order of efficiency: ATP > GTP > UTP congruent with CTP, whereas the non-hydrolyzable ATP analogue 5'-adenylyl-beta,gamma-imidodiphosphate, ADP, or PPi did not energize uptake. E217G transport into rye vacuoles was saturable (Km approximately 0.2 mM). The rye-specific luteolin glucuronides decreased uptake rates of E217G and MOC-GS into rye and barley vacuoles to comparable degrees with the mono- and diglucuronidated derivatives (40-60% inhibition) being more effective than the triglucuronide. Inhibition of E217G uptake by luteolin 7-O-diglucuronide was competitive (Ki = 120 microM). Taurocholate had no effect on E217G transport, and uptake of MOC-GS was not inhibited by E217G. Although GS conjugates and oxidized GS decreased MOC-GS transport, E217G uptake into rye and barley vacuoles was stimulated up to 7-fold in a concentration-dependent manner by these substances, with dinitrobenzene-GS being most effective. The stimulation of the GS conjugates was not due to detergent or redox effects and was specific for the E217G pump. GS conjugate stimulation of glucuronate uptake was unique for plants as E217G uptake into yeast microsomal vesicles was not affected. By comparison with a DeltaYCF1 yeast mutant, defective in vacuolar transport of GS conjugates mediated by YCF1, it was shown that E217G was taken up into yeast vesicles via a YCF1-independent directly energized pump. These results indicate that E217G as a glucuronate conjugate is transported across the vacuolar membranes of plants and yeast by a carrier distinct from the GS conjugate ATPase. PMID- 9417075 TI - Hepatocyte growth factor/scatter factor binds with high affinity to dermatan sulfate. AB - We have demonstrated by affinity chromatography that hepatocyte growth factor/scatter factor (HGF/SF) binds strongly to dermatan sulfate (DS), with a similar ionic strength dependence to that previously seen with heparan sulfate (HS). Analysis of binding kinetics on a biosensor yields an equilibrium dissociation constant, KD, of 19.7 nM. This corresponds to a 10-100-fold weaker interaction than that with HS, primarily due to a faster dissociation rate of the complex. The smallest DS oligosaccharide with significant affinity for HGF/SF by affinity chromatography appears to be an octasaccharide. A sequence comprising unsulfated iduronate residues in combination with 4-O-sulfated N acetylgalactosamine is sufficient for high affinity binding. The presence of 2-O sulfation on the iduronate residues does not appear to be inhibitory. These observations concur with our previous suggestions, from analyses of HS binding (Lyon, M., Deakin, J. A., Mizuno, K., Nakamura, T., and Gallagher, J.T. (1994) J. Biol. Chem. 269, 11216-11223), that N-sulfation of hexosamines and 2-O-sulfation of iduronates are not absolute requirements for glycosaminoglycan binding to HGF/SF. This is the first described example of a high affinity interaction between a growth factor and DS, and is likely to have significant implications for the biological activity of this paracrine-acting factor. PMID- 9417076 TI - Palmitoylation of proteolipid protein from rat brain myelin using endogenously generated 18O-fatty acids. AB - Proteolipid protein (PLP), the major protein of central nervous system myelin, contains covalently bound fatty acids, predominantly palmitic acid. This study adapts a stable isotope technique (Kuwae, T., Schmid, P. C., Johnson, S. B., and Schmid, H. O. (1990) J. Biol. Chem. 265, 5002-5007) to quantitatively determine the minimal proportion of PLP molecules which undergo palmitoylation. In these experiments, brain white matter slices from 20-day-old rats were incubated for up to 6 h in a physiological buffer containing 50% H218O. The uptake of 18O into the carbonyl groups of fatty acids derived from PLP, phospholipids, and the free fatty acid pool was measured by gas-liquid chromatography/mass spectrometry of the respective methyl esters. Palmitic acid derived from PLP acquired increasing amounts of 18O, ending with 2.9% 18O enrichment after 6 h of incubation. 18O incorporation into myelin free palmitic acid also increased over the course of the incubation (67.2% 18O enrichment). After correcting for the specific activity of the 18O-enriched free palmitic acid pool, 7.6% of the PLP molecules were found to acquire palmitic acid in 6 h. This value is not only too large to be the result of the palmitoylation of newly synthesized PLP molecules, it was also unchanged upon the inhibition of protein synthesis with cycloheximide. 18O enrichment in less actively myelinating 60-day-old rats was significantly reduced. In conclusion, our experiments suggest that a substantial proportion of PLP molecules acquire palmitic acid via an acylation/deacylation cycle and that this profile changes during development. PMID- 9417077 TI - Intermolecular exchange and stabilization of recombinant human alphaA- and alphaB crystallin. AB - Lens alpha-crystallin subunits alphaA and alphaB are differentially expressed and have a 3-to-1 ratio in most mammalian lenses by intermolecular exchange. The biological significance of this composition and the mechanism of exchange are not clear. Preparations of human recombinant alphaA- and alphaB-crystallins provide a good system in which to study this phenomenon. Both recombinant alphaA- and alphaB-crystallins are folded and aggregated to the size of the native alpha crystallin. During incubation together, they undergo an intermolecular exchange as shown by native isoelectric focusing. Circular dichroism measurements indicate that the protein with a 3-to-1 ratio of alphaA- and alphaB-crystallins has the same secondary structure but somewhat different tertiary structures after exchange: the near-UV CD increases after exchange. The resulting hybrid aggregate is more stable than the individual homogeneous aggregates: at 62 degrees C, alphaB-crystallin is more susceptible to aggregation and displays a greater light scattering than alphaA-crystallin. This heat-induced aggregation of alphaB crystallin, however, was suppressed by intermolecular exchange with alphaA crystallin. These phenomena are also observed by fast performance liquid chromatography gel filtration patterns. The protein structure of alphaB crystallin is stabilized by intermolecular exchange with alphaA-crystallin. PMID- 9417078 TI - p140Sra-1 (specifically Rac1-associated protein) is a novel specific target for Rac1 small GTPase. AB - Rac1 small GTPase plays pivotal roles in various cell functions such as cell morphology, cell polarity, and cell proliferation. We have previously identified IQGAP1 from bovine brain cytosol as a target for Rac1 by an affinity purification method. By using the same method, we purified a specifically Rac1-associated protein with a molecular mass of about 140 kDa (p140) from bovine brain cytosol. This protein interacted with guanosine 5'-(3-O-thio)triphosphate (GTPgammaS).glutathione S-transferase (GST)-Rac1 but not with the GDP.GST-Rac1, GTPgammaS.GST-Cdc42, or GTPgammaS.GST-RhoA. The amino acid sequences of this protein revealed that p140 is identified as a product of KIAA0068 gene. We denoted this protein as Sra-1 (Specifically Rac1-associated protein). Recombinant Sra-1 interacted with GTPgammaS.GST-Rac1 and weakly with GDP.Rac1 but not with GST-Cdc42 or GST-RhoA. The N-terminal domain of Sra-1 (1-407 amino acids) was responsible for the interaction with Rac1. Myc-tagged Sra-1 and the deletion mutant capable of interacting with Rac1, but not the mutants unable to bind Rac1, were colocalized with dominant active Rac1(Val-12) and cortical actin filament at the Rac1(Val-12)-induced membrane ruffling area in KB cells. Sra-1 was cosedimented with filamentous actin (F-actin), indicating that Sra-1 directly interacts with F-actin. These results suggest that Sra-1 is a novel and specific target for Rac1. PMID- 9417079 TI - Growth factor receptor-bound protein 2 SH2/SH3 domain binding to CD28 and its role in co-signaling. AB - The co-stimulatory antigen CD28 has been shown to bind to several intracellular proteins including phosphatidylinositol 3-kinase, growth factor receptor-bound protein 2 (Grb2), and ITK. Paradoxically, Grb2 and phosphatidylinositol 3-kinase binding has been mapped to a similar pYMNM motif within the CD28 cytoplasmic tail. Given the importance of CD28 co-signaling to T cell function, questions exist regarding the mechanism by which Grb2 binds to CD28, and whether the interaction plays a role in co-stimulation. To biochemically characterize Grb2/CD28 binding, we initially utilized glutathione S-transferase-Grb2 fusion proteins carrying inactivating mutations within the SH2 and SH3 domains of Grb2, and assessed their ability to bind to CD28. In vitro binding experiments indicated that the Grb2 SH2 domain is critical for the association, while the SH3 domain plays an additional role in facilitating optimal binding. Enhanced binding via the SH3 domains was not observed when the C-terminal PXXP motif within CD28 was disrupted, thereby indicating that both SH2 and SH3 domains contribute to CD28 binding. Mutations that alter Grb2 binding were found to block the CD28 dependent interleukin-2 production. Further, tyrosine phosphorylation of Vav and the costimulation-dependent activation of Jun N-terminal kinase was blocked in cells defective in CD28/Grb2 binding. These results provide evidence for an alternate CD28-mediated signaling process involving Grb2 binding to the co receptor. PMID- 9417080 TI - Expression of interleukin-10 by in vitro and in vivo activated hepatic stellate cells. AB - Activated hepatic stellate cells (HSC) participate in matrix remodeling and deposition in liver fibrosis. The present study demonstrates that interleukin (IL)-10 is expressed by HSC upon activation in vitro or in vivo and that autocrine effects of this cytokine include inhibition of collagen production. Culture activation of HSC caused a distinct increase in IL-10 mRNA level compared with freshly isolated quiescent HSC. Treatment of cultured HSC with tumor necrosis factor-alpha, transforming growth factor-beta, or lipopolysaccharide further increased IL-10 mRNA by 2-fold and resulted in the release of IL-10 protein into the medium. HSC isolated from rats after bile duct ligation (BDL) showed prominent increases in IL-10 mRNA (x 100) and protein (x 30) levels at 7 days after BDL, but such induction disappeared in advanced liver fibrosis (19 days after BDL). IL-10 expression correlated positively with mRNA expression of interstitial collagenase and inversely with that of alpha1(I) collagen. Addition of anti-IL-10 IgG to cultured HSC caused enhanced collagen production under a basal or stimulated condition with TGF-beta, tumor necrosis factor-alpha, or lipopolysaccharide. These effects were associated with increased alpha1(I) collagen mRNA and reciprocally reduced collagenase mRNA levels. Co-transfection of HSC with an IL-10 expression vector and collagen reporter genes showed a 40% inhibition of alpha1(I) collagen promoter activity. These results demonstrate that activation of HSC causes enhanced autocrine expression of IL-10 which possesses a negative autoregulatory effect on HSC collagen production mediated at least in part by alpha1(I) collagen transcriptional inhibition and stimulation of collagenase expression. These findings, along with the demonstrated early induction of HSC IL-10 expression and its late disappearance during biliary liver fibrosis, suggest its in vivo role in matrix remodeling and a possibility that failure for HSC to sustain IL-10 expression underlies pathologic progression to liver cirrhosis. PMID- 9417081 TI - RANTES and MIP-1alpha activate stats in T cells. AB - The chemokines RANTES (regulated on activation, normal T cell expressed and secreted) and MIP (macrophage inflammatory protein)-1alpha have been implicated in regulating T cell functions. RANTES-induced T cell activation is apparently mediated via two distinct signal transduction cascades: one linked to recruitment of pertussis toxin-sensitive G proteins and the other linked to protein-tyrosine kinase activation. In this report, we identified that the transcription factors Stat1 and Stat3 (for signal transducers and activators of transcription) are rapidly activated in T cells in response to RANTES and MIP-1alpha. Nuclear extracts from MOLT-4 and Jurkat T cells treated with RANTES or MIP-1alpha contain tyrosine-phosphorylated Stat1:1 and Stat1:3 dimers that exhibit DNA-binding activity. We demonstrated that RANTES and MIP-1alpha treatment of Jurkat cells resulted in transcriptional activation of a Stat-inducible gene, c-fos, with kinetics consistent with Stat activation by these chemokines. RANTES and MIP 1alpha mediate their effects via shared chemokine receptors (CCRs): CCR1, CCR4, and CCR5. Our data revealed a concordance between chemokine-induced Stat activation and c-fos induction and CCR4 and CCR5 expression. These findings indicate that chemokine-mediated activation of G-protein-coupled receptors leads to signal transduction that invokes intracellular phosphorylation intermediates used by other cytokine receptors. PMID- 9417082 TI - The Jak/Stat pathway and urokinase receptor signaling in human aortic vascular smooth muscle cells. AB - The binding of urokinase plasminogen activator (uPA) to its specific receptor (uPAR) facilitates migration of vascular smooth muscle cells (VSMC). However, the signaling cascade utilized by the urokinase receptor is only incompletely understood. We investigated intracellular uPA/uPAR signaling in human aortic VSMC from the cell membrane to the nucleus. uPA binding to VSMC induced a rapid and pronounced increase in tyrosine phosphorylation of several proteins with molecular masses of 53-60, 85-90, and 130-140 kDa. By using co immunoprecipitation techniques and in vitro kinase assays, the uPAR-associated proteins were identified as Janus (Jak) and Src non-receptor protein-tyrosine kinases (PTK) Jak1, Tyk2, and p59(fyn), p53/56(lyn), p53/59(hck), and p55(fgr). Furthermore, uPA induced a time-dependent reversible translocation of the Stat1 (signal transducer and activator of transcription) protein to the VSMC nuclei, as shown by confocal microscopy studies. Using an electrophoretic mobility shift assay, we then demonstrated that Stat1 is rapidly activated in response to stimulation with uPA and specifically binds to the DNA regulatory elements GAS (interferon-gamma activation site) and ISRE (interferon-stimulated response element). Mobility supershift experiments confirmed DNA-protein complexes containing Stat1 protein. Migration experiments with double immunofluorescence staining revealed polarization of uPAR, and colocalization with Jak1 and Tyk2 to the leading edge of the migrating cells. Under the same conditions, Jak2, Jak3, and the Src-PTKs remained randomly distributed over the entire body of the cells. Our studies therefore suggest that, in VSMC, the uPAR-signaling complex utilizes at least two different mechanisms, a direct signaling pathway utilizing the Jak/Stat cascade and a second signal transduction mechanism via Src-like protein tyrosine kinases. uPA-induced signaling via Jak/Stat is most likely involved in the regulation of cell migration, while the functional purpose of the uPA associated Src-PTK activation remains to be elucidated. PMID- 9417083 TI - Visualization of agonist-induced sequestration and down-regulation of a green fluorescent protein-tagged beta2-adrenergic receptor. AB - To date, the visualization of beta2-adrenergic receptor (beta2AR) trafficking has been largely limited to immunocytochemical analyses of acute internalization events of epitope-tagged receptors in various transfection systems. The development of a beta2AR conjugated with green fluorescent protein (beta2AR-GFP) provides the opportunity for a more extensive optical analysis of beta2AR sequestration, down-regulation, and recycling in cells. Here we demonstrate that stable expression of beta2AR-GFP in HeLa cells enables a detailed temporal and spatial analysis of these events. Time-dependent colocalization of beta2AR-GFP with rhodamine-labeled transferrin and rhodamine-labeled dextran following agonist exposure demonstrates receptor distribution to early endosomes (sequestration) and lysosomes (down-regulation), respectively. The observed temporal distribution of beta2AR-GFP was consistent with measures of receptor sequestration and down-regulation generated by radioligand-receptor binding assays. Cells stimulated with different beta-agonists revealed time courses of beta2AR-GFP redistribution reflective of the intrinsic activity of each agonist. PMID- 9417084 TI - Characterization of a fungal maleylacetoacetate isomerase gene and identification of its human homologue. AB - We have previously used Aspergillus nidulans as a fungal model for human phenylalanine catabolism. This model was crucial for our characterization of the human gene involved in alcaptonuria. We use here an identical approach to characterize at the cDNA level the human gene for maleylacetoacetate isomerase (MAAI, EC 5.2.1.2), the only as yet unidentified structural gene of the phenylalanine catabolic pathway. We report here the first characterization of a gene encoding a MAAI enzyme from any organism, the A. nidulans maiA gene. maiA disruption prevents growth on phenylalanine (Phe) and phenylacetate and results in the absence of MAAI activity in vitro and Phe toxicity. The MaiA protein shows strong amino acid sequence identity to glutathione S-transferases and has MAAI activity when expressed in Escherichia coli. maiA is clustered with fahA and hmgA, the genes encoding the two other enzymes of the common part of the Phe/phenylacetate pathways. Based on the high amino acid sequence conservation existing between other homologous A. nidulans and human enzymes of this pathway, we used the MaiA sequence in data base searches to identify human expressed sequence tags encoding its putative homologues. Four such cDNAs were sequenced and shown to be encoded by the same gene. They encode a protein with 45% sequence identity to MaiA, which showed MAAI activity when expressed in E. coli. Human MAAI deficiency would presumably cause tyrosinemia that would be characterized by the absence of succinylacetone, the diagnostic compound resulting from fumarylacetoacetate hydrolase deficiency in humans and fungi. Culture supernatants of an A. nidulans strain disrupted for maiA are succinylacetone negative but specifically contain cis and/or trans isomers of 2, 4-dioxohept-2 enoic acid. We suggest that this compound(s) might be diagnostic for human MAAI deficiency. PMID- 9417085 TI - Identification of CD44 residues important for hyaluronan binding and delineation of the binding site. AB - CD44 is a widely distributed cell surface protein that plays a role in cell adhesion and migration. As a proteoglycan, CD44 is also implicated in growth factor and chemokine binding and presentation. The extracellular region of CD44 is variably spliced, giving rise to multiple CD44 isoforms. All isoforms contain an amino-terminal domain, which is homologous to cartilage link proteins. The cartilage link protein-like domain of CD44 is important for hyaluronan binding. The structure of the link protein domain of TSG-6 has been determined by NMR. Based on this structure, a molecular model of the link-homologous region of CD44 was constructed. This model was used to select residues for site-specific mutagenesis in an effort to identify residues important for ligand binding and to outline the hyaluronan binding site. Twenty-four point mutants were generated and characterized, and eight residues were identified as critical for binding or to support the interaction. In the model, these residues form a coherent surface the location of which approximately corresponds to the carbohydrate binding sites in two functionally unrelated calcium-dependent lectins, mannose-binding protein and E-selectin (CD62E). PMID- 9417086 TI - Rapid redistribution of CD20 to a low density detergent-insoluble membrane compartment. AB - CD20 is a B cell integral membrane protein capable of initiating growth modulating signals in human B lymphocytes upon its engagement with monoclonal anti-CD20 antibodies. In this report, we demonstrate that treatment of B cells with CD20 antibodies induces rapid redistribution of CD20 into a detergent insoluble membrane compartment. Redistribution is detected as early as 15 s, following antibody addition, and involves up to 95% of CD20 molecules, depending on the antibody used. All of the detergent-insoluble CD20 was found in the low density fractions of sucrose density gradients, indicating that CD20 redistributes to glycolipid-rich membrane domains, analogous to caveolae in some cell types. As CD20 has previously been shown to associate with Src family tyrosine kinases, their co-existence in these compartments suggests a link to the role of CD20 in signal transduction. This study provides insight into the mechanism by which CD20 communicates signals to the cell interior and indicates that the search for membrane-proximal intracellular signaling partners should be directed to the Triton-insoluble fraction. PMID- 9417087 TI - Delta3,5-delta2,4-dienoyl-CoA isomerase from rat liver. Molecular characterization. AB - rECH1, a recently identified rat cDNA (FitzPatrick, D. R., Germain-Lee, E., and Valle, D. (1995) Genomics 27, 457-466) encodes a polypeptide belonging to the hydratase/isomerase superfamily. We modeled the structure of rECH1 based on rat mitochondrial 2-enoyl-CoA hydratase 1. The model predicts that rECH1p has the hydratase fold in the core domain and two domains for interaction with other subunits. When we incubated 3,5,8,11, 14-eicosapentaenoyl-CoA with purified rECH1p, the spectral data suggested a switching of the double bonds from the Delta3-Delta5 to the Delta2-Delta4 positions. This was confirmed by demonstrating that the product was a valid substrate for 2,4-dienoyl-CoA reductase. These results indicate that rECH1p is Delta3,5-Delta2,4-dienoyl-CoA isomerase. Subcellular fractionation and immunoelectron microscopy using antibodies to a synthetic polypeptide derived from the C terminus of rECH1p showed that rECH1p is located in the matrix of both mitochondria and peroxisomes in rat liver. Consistent with these observations, the 36,000-Da rECH1p has a potential N terminal mitochondrial targeting signal as well as a C-terminal peroxisomal targeting signal type 1. Transport of the protein into the mitochondria with cleavage of the targeting signal results in a mature mitochondrial form with a molecular mass of 32,000 Da; transport to peroxisomes yields a protein of 36,000 Da. PMID- 9417088 TI - Solution structure of Der f 2, the major mite allergen for atopic diseases. AB - House dust mites cause heavy atopic diseases such as asthma and dermatitis. Among allergens from Dermatophagoides farinae, Der f 2 shows the highest positive rate for atopic patients, but its biological function in mites has been perfectly unknown, as well as the functions of its homologs in human and other animals. We have determined the tertiary structure of Der f 2 by multidimensional nuclear magnetic resonance spectroscopy. Der f 2 was found to be a single-domain protein of immunoglobulin fold, and its structure was the most similar to those of the two regulatory domains of transglutaminase. This fact, binding to the bacterial surface, and other small pieces of information hinted that Der f 2 is related to the innate antibacterial defense system in mites. The immunoglobulin E epitopes are also discussed on the basis of the tertiary structure. PMID- 9417089 TI - Molecular cloning, sequence analysis, expression, and tissue distribution of suppressin, a novel suppressor of cell cycle entry. AB - Suppressin (SPN) is an inhibitor of cell proliferation that was originally identified and purified to homogeneity from bovine pituitaries (LeBoeuf, R. D., Burns, J. N., Bost, K. L., and Blalock, J. E. (1990) J. Biol. Chem. 265, 158 165). In this report we have cloned the full-length cDNA encoding rat SPN and have identified the tissue distribution of SPN expression. The cDNA of SPN is 1882 nucleotides with a 1488-base coding region and 55 and 339 nucleotides of 5'- and 3'-untranslated sequences, respectively. Northern gel analysis of rat pituitary mRNA showed a single hybridizing species at approximately 2 kilobases. Sequence analyses showed that the nucleotide and deduced amino acid sequences of SPN are novel and unrelated to any known vertebrate inhibitors of proliferation. However, the deduced amino acid sequence of SPN contains two domains that have extensive sequence identity with a recently cloned transcription activator in Drosophila, deformed epidermal autoregulatory factor-1 (DEAF-1, see Gross, C. T., and McGinnis, W. (1996) EMBO J. 15, 1961-1970) suggesting that SPN represents a vertebrate cognate of deformed epidermal autoregulatory factor-1. Reverse transcriptase-polymerase chain reaction and immunohistochemical analyses showed that the SPN mRNA and the SPN protein are expressed in every tissue examined including testis, spleen, skeletal muscle, liver, kidney, heart, and brain suggesting that SPN may be involved in the control of proliferation in a variety of cell types. PMID- 9417090 TI - Molecular cloning and characterization of a Drosophila p38 mitogen-activated protein kinase. AB - A mitogen-activated protein kinase (MAPK) has been cloned and sequenced from a Drosophila neoplasmic l(2)mbn cell line. The cDNA sequence analysis showed that this Drosophila kinase is a homologue of mammalian p38 MAPK and the yeast HOG1 gene and thus was referred to as Dp38. A distinguishing feature of all MAPKs is the conserved sequence TGY in the activation domain. Dp38 was rapidly tyrosine 186-phosphorylated in response to osmotic stress, heat shock, serum starvation, and H2O2 in Drosophila l(2)mbn and Schneider cell lines. However, unlike mammalian p38 MAPK, the addition of lipopolysaccharide (LPS) did not significantly affect the phosphorylation of Dp38 in the LPS-responsive l(2)mbn cell line. Following osmotic stress, tyrosine 186-phosphorylated forms of Dp38 MAPK were detected exclusively in nuclear regions of Schneider cells. Yeast complementation studies demonstrated that the Saccharomyces cerevisiae HOG1 mutant strain JBY10 (hog1-Delta1) was functionally complemented by Dp38 cDNA in hyperosmolar medium. These findings demonstrate that similar osmotic stress responsive signal transduction pathways are conserved in yeast, Drosophila, and mammalian cells, whereas LPS signal transduction pathways appear to be different. PMID- 9417091 TI - Syntaxin 7, a novel syntaxin member associated with the early endosomal compartment. AB - Members of the syntaxin family are key molecules involved in diverse vesicle docking/fusion events. We report here the molecular, biochemical, and cell biological characterizations of a novel member (syntaxin 7) of the syntaxin family. Syntaxin 7 is structurally related to all known syntaxins. Within a 79 residue region preceding the C-terminal hydrophobic tail, syntaxin 7 is 35, 34, 34, 34, 25, and 19% identical to syntaxins 1, 2, 3, 4, 5, and 6, respectively. Northern blot analysis showed that syntaxin 7 is widely expressed. Indirect immunofluorescence microscopy revealed that syntaxin 7 is primarily associated with the early endosome. In vitro binding assays established that syntaxin 7 in membrane extracts interacts with immobilized recombinant alpha-soluble N ethylmaleimide-sensitive factor attachment proteins fused to glutathione S transferase. Our results highlight the general importance of members of the syntaxin family in protein trafficking and provide new avenues for future functional and mechanistic studies of this first endosomal syntaxin as well as the endocytotic pathway. PMID- 9417092 TI - X-gene product of hepatitis B virus induces apoptosis in liver cells. AB - Hepatitis B virus is a causative agent of hepatocellular carcinoma, and in the course of tumorigenesis, the X-gene product (HBx) is known to play important roles. Here, we investigated the transforming potential of HBx by conventional focus formation assay in NIH3T3 cells. Cells were cotransfected with the HBx expression plasmid along with other oncogenes including Ha-ras, v-src, v-myc, v fos, and E1a. Unexpectedly, the introduction of HBx completely abrogated the focus-forming ability of all five tested oncogenes. In addition, the cotransfection of Bcl-2, an apoptosis inhibitor, reversed the HBx-mediated inhibition of focus formation, suggesting that the observed repression of focus formation by HBx is through the induction of apoptosis. Next, to test unequivocally whether HBx induces apoptosis in liver cells, we established stable Chang liver cell lines expressing HBx under the control of a tetracycline inducible promoter. Induction of HBx in these cells in the presence of 1% calf serum resulted in typical apoptosis phenomena such as DNA fragmentation, nuclear condensation, and fragmentation. Based on these results, we propose that HBx sensitizes liver cells to apoptosis upon hepatitis B virus infection, contributing to the development of hepatitis and the subsequent generation of hepatocellular carcinoma. PMID- 9417093 TI - The CC chemokine I-309 inhibits CCR8-dependent infection by diverse HIV-1 strains. AB - Using a chemokine receptor model based on known receptor sequences, we identified several members of the seven transmembrane domain G-protein superfamily as potential chemokine receptors. The orphan receptor ChemR1, which has recently been shown to be a receptor for the CC chemokine I-309, scored very high in our model. We have confirmed that I-309, but not a number of other chemokines, can induce a transient Ca2+ flux in cells expressing CCR8. In addition, the human erythroleukemic cell line K562 responded chemotactically in a dose-responsive manner to this chemokine. Since several chemokine receptors have been shown to be required as coreceptors for HIV-1 infection, we asked whether human immunodeficiency virus type 1 (HIV-1) could efficiently utilize CCR8. Here we show that the CCR8 receptor can serve as a coreceptor for diverse T-cell tropic, dual-tropic, and macrophage-tropic HIV-1 strains and that I-309 was a potent inhibitor of HIV-1 envelope-mediated cell-cell fusion and virus infection. Furthermore, we show by flow cytometry and immunohistochemistry that antibodies generated against the CCR8 receptor amino-terminal peptide cross-reacted with U 87 MG cells stably expressing CCR8, THP-1 cells, HL-60 cells, and human monocytes, a target cell for HIV-1 infectivity in vivo. PMID- 9417094 TI - Thioltransferase (glutaredoxin) reactivates the DNA-binding activity of oxidation inactivated nuclear factor I. AB - The reversible oxidative inactivation of transcription factors has been proposed to be important in cellular responses to oxidant stress and in several signal transduction pathways. The nuclear factor I (NFI) family of transcription factors is sensitive to oxidative inactivation due to the presence of a conserved, oxidation-sensitive cysteine residue within the NFI DNA-binding domain. Here we show that restoration of the DNA-binding activity of oxidized NFI-C can be catalyzed in vitro by the cellular enzyme thioltransferase (glutaredoxin) coupled to GSH and GSSG reductase. To test whether GSH-dependent pathways play a role in the maintenance of NFI activity in vivo, we used buthionine sulfoximine, an agent that inhibits GSH synthesis, and N-acetylcysteine, an agent that can replenish intracellular GSH. Pretreatment of HeLa cells with buthionine sulfoximine greatly potentiated the inactivation of NFI by the oxidizing agent diamide. Inclusion of N-acetylcysteine in the culture medium during the recovery period following diamide treatment increased the extent of restoration of NFI activity. These results suggest that maintenance of the DNA-binding activity of NFI proteins during oxidant stress in vivo requires a GSH-dependent pathway, likely involving thioltransferase-catalyzed reduction of the oxidation-sensitive cysteine residue on NFI. PMID- 9417095 TI - Identification of an inhibin receptor in gonadal tumors from inhibin alpha subunit knockout mice. AB - Inhibins and activins are dimeric proteins that are functional antagonists and are structurally related to the transforming growth factor-beta (TGFbeta) family of growth and differentiation factors. Receptors for activin and TGFbeta have been identified as dimers of serine-threonine kinase subunits that regulate cytoplasmic proteins known as Smads. Despite major advances in our understanding of activin and TGFbeta receptors and signaling pathways, little is known about inhibin receptors or the mechanism by which this molecule provides a functionally antagonistic signal to activin. Studies described in this paper indicate that an independent inhibin receptor exists. Numerous tissues were examined for inhibin specific binding sites, including the developing embryo, in which the spinal ganglion and trigeminal ganglion-bound iodinated inhibin A. Sex cord stromal tumors, derived from male and female inhibin alpha-subunit-deficient mice, were also identified as a source of inhibin receptor. Abundant inhibin and few activin binding sites were identified in tumor tissue sections by in situ ligand binding using iodinated recombinant human inhibin A and 125I-labeled recombinant human inhibin A. Tumor cell binding was specific for each ligand (competed by excess unlabeled homologous ligand and not competed by heterologous ligand). Based on these results and the relative abundance and homogeneity of tumor tissues versus the embryonic ganglion, tumor tissues were homogenized, membrane proteins were purified, and putative inhibin receptors were isolated using an inhibin affinity column. Four proteins were eluted from the column that bind iodinated inhibin but not iodinated activin. These data suggest that inhibin-specific membrane associated proteins (receptors) exist. PMID- 9417097 TI - Protein kinase D activation by mutations within its pleckstrin homology domain. AB - Protein kinase D (PKD) is a serine/threonine protein kinase that contains a cysteine-rich repeat sequence homologous to that seen in the regulatory domain of protein kinase C (PKC) and a catalytic domain with only a low degree of sequence similarity to PKCs. PKD also contains a pleckstrin homology (PH) domain inserted between the cysteine-rich motifs and the catalytic domain that is not present in any of the PKCs. To investigate the function of the PH domain in the regulation of PKD activity, we determined the kinase activity of several PKD PH domain mutants immunoprecipitated from lysates of transiently transfected COS-7 cells. Deletion of the entire PH domain (amino acids 429-557) markedly increased the basal activity of the enzyme as assessed by autophosphorylation ( approximately 16-fold) and exogenous syntide-2 peptide substrate phosphorylation assays (approximately 12-fold). Mutant PKD proteins with partial deletions or single amino acid substitutions within the PH domain (e. g. R447C and W538A) also exhibited increased basal kinase activity. These constitutive active mutants of PKD were only slightly further stimulated by phorbol-12,13-dibutyrate treatment of intact cells. Our results demonstrate, for the first time, that the PKD PH domain plays a negative role in the regulation of enzyme activity. PMID- 9417096 TI - Conformational changes in cell surface HIV-1 envelope glycoproteins are triggered by cooperation between cell surface CD4 and co-receptors. AB - We have continuously measured CD4-induced conformational changes of cell surface expressed human immunodeficiency virus type-1 envelope glycoprotein gp120-gp41 in situ using 4,4'-dianilino-1, 1'-binaphthyl-5,5'-disulfonic acid, a fluorescent probe that binds to hydrophobic groups. CD4-expressing human T cell lines induced significant and rapid conformational changes (<1 min delay) in gp120-gp41 from T cell-tropic strains, and little conformational changes in gp120-gp41 from macrophage-tropic strains, with equivalent levels of envelope expression. Conversely, CD4-expressing human macrophages induced significant and rapid conformational changes in gp120-gp41 from macrophage-tropic strains, and little conformational changes in gp120-gp41 from T cell-tropic strains. Thus, the conformational changes undergone by gp120-gp41, which lead to membrane fusion, are highly cooperative and require both receptor and co-receptor. We used a dye transfer assay to show that neither membrane lipid fusion or fusion pore formation can occur with host cells having different tropism from the envelope. PMID- 9417098 TI - Replacements of single basic amino acids in the pleckstrin homology domain of phospholipase C-delta1 alter the ligand binding, phospholipase activity, and interaction with the plasma membrane. AB - The pleckstrin homology (PH) domain of phosphatidylinositol-specific phospholipase C-delta1 (PLC-delta1) binds to both D-myo-inositol 1,4, 5 trisphosphate (Ins(1,4,5)P3) and phosphatidylinositol 4, 5-bisphosphate (PtdIns(4,5)P2) with high affinities. We have previously identified a region rich in basic amino acids within the PH domain critical for ligand binding (Yagisawa, H., Hirata, M., Kanematsu, T., Watanabe, Y., Ozaki, S., Sakuma, K., Tanaka, H., Yabuta, N., Kamata, H., Hirata, H., and Nojima, H. (1994) J. Biol. Chem. 269, 20179-20188; Hirata, M., Kanematsu, T., Sakuma, K., Koga, T., Watanabe, Y., Ozaki, S., and Yagisawa, H. (1994) Biochem. Biophys. Res. Commun. 205, 1563 1571). To investigate the role of these basic residues, we have performed site directed mutagenesis replacing each of the basic amino acid in the N-terminal 60 residues of PLC-delta1 (Lys24, Lys30, Lys32, Arg37, Arg38, Arg40, Lys43, Lys49, Arg56, Lys57, and Arg60) with a neutral or an acidic amino acid. The effects of these mutations on the PH domain ligand binding properties and their consequence for substrate hydrolysis and membrane interactions of PLC-delta1 were analyzed using several assay systems. Analysis of [3H]-Ins(1,4,5)P3 binding, measurement of the binding affinities, and measurements of phospholipase activity using PtdIns(4,5)P2-containing phospholipid vesicles, demonstrated that residues Lys30, Lys32, Arg37, Arg38, Arg40, and Lys57 were required for these PLC-delta1 functions; in comparison, other mutations resulted in a moderate reduction. A subset of selected mutations was further analyzed for the enzyme activity toward substrate present in cellular membranes of permeabilized cells and for interaction with the plasma membrane after microinjection. These experiments demonstrated that mutations affecting ligand binding and PtdIns(4,5)P2 hydrolysis in phospholipid vesicles also resulted in reduction in the hydrolysis of cellular polyphosphoinositides and loss of membrane attachment. All residues (with the exception of the K43E substitution) found to be critical for the analyzed PLC delta1 functions are present at the surface of the PH domain shown to contain the Ins(1,4,5)P3 binding pocket. PMID- 9417099 TI - A cluster of basic residues in the carboxyl-terminal tail of the short metabotropic glutamate receptor 1 variants impairs their coupling to phospholipase C. AB - Among phospholipase C-coupled metabotropic glutamate receptors (mGluRs), some have a surprisingly long carboxyl-terminal intracellular domain (mGluR1a, -5a, and -5b), and others have a short one (mGluR1b, -1c, and -1d). All mGluR1 sequences are identical up to 46 residues following the 7th transmembrane domain, followed by 313, 20, 11, and 26 specific residues in mGluR1a, mGluR1b, mGluR1c, and mGluR1d, respectively. Several functional differences have been described between the long isoforms (mGluR1a, -5a, and -5b) and the short ones (mGluR1b, 1c, and -1d). Compared with the long receptors, the short ones induce slower increases in intracellular Ca2+, are activated by higher concentration of agonists, and do not exhibit constitutive, agonist-independent activity. To identify the residues responsible for these functional properties, a series of truncated, chimeric, and mutated receptors were constructed. We found that the deletion of the last 19 carboxyl-terminal residues in mGluR1c changed its properties into those of mGluR1a. Moreover, the exchange of the long carboxyl terminal domain of mGluR5a with that of mGluR1c generated a chimeric receptor that possessed functional properties similar to those of mGluR1c. Mutagenesis of specific residues within the 19 carboxyl-terminal residues of mGluR1c revealed the importance of a cluster of 4 basic residues in defining the specific properties of this receptor. Since this cluster is part of the sequence common to all mGluR1 variants, we conclude that the long carboxyl-terminal domain of mGluR1a suppresses the inhibitory action of this sequence element. PMID- 9417100 TI - Cloning of a human UDP-galactose:2-acetamido-2-deoxy-D-glucose 3beta galactosyltransferase catalyzing the formation of type 1 chains. AB - Biochemical evidence suggests that the galactosyltransferase activity synthesizing type 1 carbohydrate chains is separate from the well characterized enzyme that is responsible for the synthesis of type 2 chains. This was recently confirmed by the cloning, from melanoma cells, of an enzyme capable of synthesizing type 1 chains, which was shown to have no homology to other galactosyltransferases. We report here the molecular cloning and functional expression of a second human beta3-galactosyltransferase distinct from the melanoma enzyme. The new beta3-galactosyltransferase has homology to the melanoma enzyme in the putative catalytic domain, but has longer cytoplasmic and stem regions and a carboxyl-terminal extension. Northern blots showed that the new gene is present primarily in brain and heart. When transfected into mammalian cells, this gene directs the synthesis of type 1 chains as determined by a monoclonal antibody specific for sialyl Lewisa. A soluble version of the cloned enzyme was expressed in insect cells and purified. The soluble enzyme readily catalyzes the transfer of galactose to GlcNAc to form Gal(beta1-3)GlcNAc. It also has a minor but distinct transfer activity toward Gal, LacNAc, and lactose, but is inactive toward GalNAc. PMID- 9417101 TI - Essential requirement of cytosolic phospholipase A2 for activation of the phagocyte NADPH oxidase. AB - Arachidonic acid (AA) can trigger activation of the phagocyte NADPH oxidase in a cell-free assay. However, a role for AA in activation of the oxidase in intact cells has not been established, nor has the AA generating enzyme critical to this process been identified. The human myeloid cell line PLB-985 was transfected to express p85 cytosolic phospholipase A2 (cPLA2) antisense mRNA and stable clones were selected that lack detectable cPLA2. cPLA2-deficient PLB-985 cells differentiate similarly to control PLB-985 cells in response to retinoic acid or 1,25-dihydroxyvitamin D3, indicating that cPLA2 is not involved in the differentiation process. Neither cPLA2 nor stimulated [3H]AA release were detectable in differentiated cPLA2-deficient PLB-985 cells, demonstrating that cPLA2 is the major type of PLA2 activated in phagocytic-like cells. Despite the normal synthesis of NADPH oxidase subunits during differentiation of cPLA2 deficient PLB-985 cells, these cells fail to activate NADPH oxidase in response to a variety of soluble and particulate stimuli, but the addition of exogenous AA fully restores oxidase activity. This establishes an essential requirement of cPLA2-generated AA for activation of phagocyte NADPH oxidase. PMID- 9417103 TI - Purification and characterization of procytotoxin of Pseudomonas aeruginosa. Dimer to monomer conversion of protoxin by proteolytic activation. AB - Cytotoxin of Pseudomonas aeruginosa is a cytolytic toxin that forms a pore on the target membrane by oligomerizing into a pentamer. This toxin is produced as an inactive precursor (proCTX) and is converted to an active form by proteolytic cleavage at the C terminus. We purified proCTX to apparent homogeneity and characterized it in a comparison with the active toxin. ProCTX bound to the erythrocyte membrane but did not form an oligomer on the membrane, hence the lack of hemolytic activity in proCTX. Circular dichroic experiments showed that active and proCTX have similar beta-sheet dominant structures. Intrinsic fluorescence analysis indicated that a molecule-buried tryptophan residue(s) of proCTX was exposed to the surface of the molecule as a result of conversion to the active form. In analytical gel filtration, chemical cross-linking, and analytical ultracentrifugation experiments, dimer to monomer conversion occurred with proteolytic activation. PMID- 9417102 TI - Glyceraldehyde-3-phosphate dehydrogenase is regulated on a daily basis by the circadian clock. AB - Circadian clocks function to govern a wide range of rhythmic activities in organisms. An integral part of rhythmicity is the daily control of target genes by the clock. Here we describe the sequence and analysis of a novel clock controlled gene, ccg-7, showing similarity to glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a glycolytic enzyme widely used as a constitutive control in a variety of systems. That ccg-7 encodes GAPDH was confirmed by demonstrating that in vitro synthesized CCG-7 possesses GAPDH activity. Rhythms in both ccg-7 mRNA accumulation and CCG-7 (GAPDH) activity are observed in a clock wild-type strain where the peak in GAPDH activity lags several hours behind the peak in ccg 7 mRNA accumulation in the late night. Together with our previous observation that ccg-7 mRNA is not developmentally regulated, we show that ccg-7 is not induced by environmental stresses such as glucose or nitrogen deprivation (which also trigger development), heat shock, or osmotic stress. Thus, the finding that GAPDH is clock-regulated points to a specific role for the circadian clock in controlling aspects of general metabolism and provides evidence for circadian regulation of a gene found in most living organisms. PMID- 9417104 TI - Versatile action of Escherichia coli ClpXP as protease or molecular chaperone for bacteriophage Mu transposition. AB - The molecular chaperone ClpX of Escherichia coli plays two distinct functions for bacteriophage Mu DNA replication by transposition. As specificity component of a chaperone-linked protease, it recognizes the Mu immunity repressor for degradation by the peptidase component ClpP, thus derepressing Mu transposition functions. After strand exchange has been promoted by MuA transposase, ClpX alone can alter the conformation of the transpososome (the complex of MuA with Mu ends), and the remodeled MuA promotes transition to replisome assembly. Although ClpXP can degrade MuA, the presence of both ClpP and ClpX in the reconstituted transposition system did not destroy MuA essential for initiation of DNA replication by specific host replication enzymes. Levels of ClpXP needed to overcome inhibition by the repressor did not prevent MuA from promoting strand transfer, and ClpP stimulated alteration of the transpososome by ClpX. Apparently intact MuA was still present in the resulting transpososome, promoting initiation of Mu DNA replication by specific replication enzymes. The results indicate that ClpXP can discriminate repressor and MuA in the transpososome as substrates of the protease or the molecular chaperone alone, degrading repressor while remodeling MuA for its next critical function. PMID- 9417105 TI - Structural elucidation and monokine-inducing activity of two biologically active zwitterionic glycosphingolipids derived from the porcine parasitic nematode Ascaris suum. AB - The isolated neutral glycosphingolipid fraction from the pig parasitic nematode, Ascaris suum, was fractionated by silica gel chromatography to yield a neutral and a zwitterionic glycosphingolipid fraction, the latter of which mainly contained two zwitterionic glycosphingolipids termed components A and C. Preliminary chemical characterization with hydrofluoric acid treatment and immunochemical characterization with a phosphocholine-specific monoclonal antibody indicated that both components contained phosphodiester substitutions: phosphocholine for component A, and phosphocholine and phosphoethanolamine for component C. Both components were biologically active in inducing human peripheral blood mononuclear cells to release the inflammatory monokines tumor necrosis factor alpha, interleukin 1, and interleukin 6. Component A was the more bioactive molecule, and its biological activity was abolished on removal of the phosphocholine substituent by hydrofluoric acid. The glycosphingolipid components were structurally analyzed by matrix-assisted laser desorption/ionization time-of flight mass spectrometry, liquid secondary ion mass spectrometry, methylation analysis, 1H NMR spectroscopy, exoglycosidase cleavage, and ceramide analysis. Their chemical structures were elucidated to be (see Structure I below), [structure: see text] The carbohydrate moiety oligosaccharide core was characterized as belonging to the arthro series of protostomial glycosphingolipids. The ceramide moiety was distinguished by (R)-2 hydroxytetracosanoic acid as the dominant fatty acid species and by the C17 iso branched sphingosine and sphinganine bases, 15-methylhexadecasphing-4-enine and 15-methylhexadecasphinganine, respectively. PMID- 9417106 TI - The identification and characterization of oligodendrocyte thromboxane A2 receptors. AB - The presence of functional thromboxane A2 receptors in neonatal rat oligodendrocytes and human oligodendroglioma cells was investigated using immunocytochemistry, ligand affinity chromatography, radioligand binding analysis, immunoblot analysis, and calcium mobilization studies. Immunocytochemical studies revealed the presence of receptor protein on both oligodendrocytes and human oligodendroglioma cells. Ligand affinity chromatography allowed for the purification of a protein with an electrophoretic mobility (55 kDa) indistinguishable from human platelet thromboxane A2 receptors. This affinity purified protein was immunoreactive against a polyclonal anti thromboxane A2 receptor antibody. Intact human oligodendroglioma cells specifically bound [3H]SQ29,548 with a KD of 4 nM and were found to have approximately 3500 binding sites per cell. Human oligodendroglioma cells also demonstrated calcium mobilization in response to receptor activation with U46619. These results demonstrate the presence of a functional thromboxane A2 receptor in oligodendrocytes and are consistent with previous observations indicating a high density of thromboxane A2 receptors in myelinated brain and spinal cord fiber tracts. PMID- 9417107 TI - Negative regulation of beta enolase gene transcription in embryonic muscle is dependent upon a zinc finger factor that binds to the G-rich box within the muscle-specific enhancer. AB - We have previously identified a muscle-specific enhancer within the first intron of the human beta enolase gene. Present in this enhancer are an A/T-rich box that binds MEF-2 protein(s) and a G-rich box (AGTGGGGGAGGGGGCTGCG) that interacts with ubiquitously expressed factors. Both elements are required for tissue-specific expression of the gene in skeletal muscle cells. Here, we report the identification and characterization of a Kruppel-like zinc finger protein, termed beta enolase repressor factor 1, that binds in a sequence-specific manner to the G-rich box and functions as a repressor of the beta enolase gene transcription in transient transfection assays. Using fusion polypeptides of beta enolase repressor factor 1 and the yeast GAL4 DNA-binding domain, we have identified an amino-terminal region responsible for the transcriptional repression activity, whereas a carboxyl-terminal region was shown to contain a potential transcriptional activation domain. The expression of this protein decreases in developing skeletal muscles, correlating with lack of binding activity in nuclear extract from adult skeletal tissue, in which novel binding activities have been detected. These results suggest that in addition to the identified factor, which functionally acts as a negative regulator and is enriched in embryonic muscle, the G-rich box binds other factors, presumably exerting a positive control on transcription. The interplay between factors that repress or activate transcription may constitute a developmentally regulated mechanism that modulates beta enolase gene expression in skeletal muscle. PMID- 9417109 TI - Characteristics of the intron involvement in the mitogen-induced expression of Zfp-36. AB - Zfp-36, the gene encoding the putative zinc finger protein tristetraprolin (TTP), is rapidly induced in fibroblasts by a variety of growth factors. Recent gene knockout experiments have shown that TTP-deficient mice developed arthritis, cachexia, and autoimmunity, all apparently mediated by an excess of tumor necrosis factor alpha. We recently showed that full serum inducibility of Zfp-36 requires elements in the promoter; in addition, removal of the single intron strikingly inhibited serum-induced TTP expression. We show here that replacement of the intron with unrelated sequences, or removal of 95% of the intron but retention of the splice sites, each resulted in the maintenance of approximately 45 and 19%, respectively, of full serum-induced expression. In addition, deletion of intron sequences base pairs 601-655 decreased the serum-induced expression of TTP by 65%. Sequence base pairs 618-626 bound specifically to the transcription factor Sp1; mutation of this binding motif decreased TTP expression by 70%, suggesting that Sp1 binding to this motif contributes to serum induction of Zfp 36. We conclude that full serum-induced expression of Zfp-36 depends on the activation of conventional promoter elements as well as elements in the single intron, and that the presence per se of the intron in its natural location also contributes significantly to the regulated expression of this gene. PMID- 9417108 TI - Interactions with single-stranded and double-stranded DNA-binding factors and alternative promoter conformation upon transcriptional activation of the Htf9 a/RanBP1 and Htf9-c genes. AB - The murine Htf9-a/RanBP1 and Htf9-c genes are divergently transcribed from a shared TATA-less promoter. Transcription of both genes is initiated on complementary DNA strands and is controlled by cell cycle-dependent mechanisms. The bidirectional promoter harbors a genomic footprint flanking the major transcription start site of both genes. Transient promoter assays showed that the footprinted element is important for transcription of both genes. Protein-binding experiments and antibody assays indicated that members of the retinoid X receptor family interact with the double-stranded site. In addition, distinct factors interact with single DNA strands of the element. Double-stranded binding factors were highly expressed in liver cells, in which neither gene is transcribed, while single-stranded binding proteins were abundant in cycling cells, in which transcription of both genes is efficient. In vivo S1 analysis of the promoter depicted an S1-sensitive organization in cells in which transcription of both genes is active; S1 sensitivity was not detected in conditions of transcriptional repression. Thus, the same element is a target for either retinoid X receptor factors, or for single-stranded binding proteins, and form distinct complexes in different cellular conditions depending on the DNA conformation in the binding site. PMID- 9417110 TI - Isolation and characterization of the phage T4 PinA protein, an inhibitor of the ATP-dependent lon protease of Escherichia coli. AB - The bacteriophage T4 PinA protein, expression of which leads to inhibition of protein degradation in Escherichia coli cells, has been purified from cells carrying multiple copies of the pinA gene. PinA is a heat-stable protein with a subunit Mr of 18,800 and an isoelectric point of 4.6. Under nondenaturing conditions on a gel filtration column, PinA migrated in two peaks corresponding to a dimer and a tetramer. Purified PinA inhibited ATP-dependent protein degradation by Lon protease in vitro; it did not inhibit the activity of other E. coli ATP-dependent proteases, ClpAP or ClpYQ. Furthermore, PinA did not inhibit ATP-independent proteolysis in E. coli cell extracts. PinA binds with high affinity to Lon protease (Kd approximately 10 nM for dimer binding), and a complex with approximately 1 dimer of PinA per tetramer of Lon protease could be isolated by gel filtration. Lon activity was partially restored upon dilution of the PinA-Lon complex to subnanomolar concentrations, indicating that inhibition was reversible and that PinA did not covalently modify Lon protease. PinA was not cleaved by Lon protease, and heating the Lon-PinA complex at 65 degrees C denatured Lon protease and released active PinA. The properties of PinA in vitro suggest that PinA inhibits protein degradation in vivo by forming a tight, reversible complex with Lon protease. PMID- 9417111 TI - PinA inhibits ATP hydrolysis and energy-dependent protein degradation by Lon protease. AB - The bacteriophage T4 PinA protein inhibited degradation of [3H]alpha-methyl casein by purified Lon protease from Escherichia coli, but inhibition was noncompetitive with respect to casein. PinA did not inhibit cleavage of the fluorogenic peptide, N-glutaryl-alanylalanylphenylalanyl-3-methoxynaphthylamide and, moreover, did not block the ability of protein substrates, such as casein, to activate cleavage of fluorogenic peptides by Lon. Thus, PinA does not block the proteolytic active site or the allosteric protein-binding site on Lon. Inhibition of basal ATPase activity was variable (50-90%), whereas inhibition of protein-activated ATPase activity was usually 80-95%. Inhibition was noncompetitive with respect to ATP. PinA did not block activation of peptide cleavage by nonhydrolyzable analogs of ATP. These data suggest that PinA does not bind at the ATPase active site of Lon and does not interfere with nucleotide binding to the enzyme. PinA inhibited cleavage of the 72-amino acid protein, CcdA, degradation of which requires ATP hydrolysis, but did not inhibit cleavage of the carboxyl-terminal 41-amino acid fragment of CcdA, degradation of which does not require ATP hydrolysis. PinA thus appears to interact at a novel regulatory or enzymatic site involved in the coupling between ATP hydrolysis and proteolysis, possibly blocking the protein unfolding or remodeling step essential for degradation of high molecular weight protein substrates by Lon. PMID- 9417112 TI - Integrin-linked protein kinase regulates fibronectin matrix assembly, E-cadherin expression, and tumorigenicity. AB - Fibronectin (Fn) matrix plays important roles in many biological processes including morphogenesis and tumorigenesis. Recent studies have demonstrated a critical role of integrin cytoplasmic domains in regulating Fn matrix assembly, implying that intracellular integrin-binding proteins may be involved in controlling extracellular Fn matrix assembly. We report here that overexpression of integrin-linked kinase (ILK), a newly identified serine/threonine kinase that binds to the integrin beta1 cytoplasmic domain, dramatically stimulated Fn matrix assembly in epithelial cells. The integrin-linked kinase activity is involved in transducing signals leading to the up-regulation of Fn matrix assembly, as overexpression of a kinase-inactive ILK mutant failed to enhance the matrix assembly. Moreover, the increase in Fn matrix assembly induced by ILK overexpression was accompanied by a substantial reduction in the cellular E cadherin. Finally, we show that ILK-overexpressing epithelial cells readily formed tumors in nude mice, despite forming an extensive Fn matrix. These results identify ILK as an important regulator of pericellular Fn matrix assembly, and suggest a novel critical role of this integrin-linked kinase in cell growth, cell survival, and tumorigenesis. PMID- 9417113 TI - JunB is involved in the inhibition of myogenic differentiation by bone morphogenetic protein-2. AB - Bone morphogenetic proteins (BMPs) constitute a family of multifunctional growth and differentiation factors structurally related to transforming growth factor beta. BMPs were first identified by their osteoinductive effects, inducing ectopic bone formation when implanted in skeletal muscle, and have an important role as regulators of skeletal development in vivo. In vitro, BMP-2 is able to transdifferentiate myogenic C2C12 cells into the osteoblastic phenotype. In this report, we show that the osteoinductive effects of BMP-2 in C2C12 cells are mediated by bone morphogenetic protein receptor type-IA in combination with both activin receptor type II and bone morphogenetic protein receptor type II. We also analyzed the expression levels of nuclear protooncogenes to understand early transcriptional events induced by BMP-2. We show that junB is an immediate early gene induced by BMP-2 and transforming growth factor-beta. BMP-2 induces transcriptional activation of JunB expression as early as 30 min after ligand addition, reaching maximal levels after 90 min. Increase of JunB mRNA correlates with a higher AP-1 binding activity. Furthermore, ectopic overexpression of JunB is sufficient to inhibit expression of myoblast differentiation markers in C2C12 cells. These data, taken together, show the involvement of JunB in the early steps of inhibition of myogenic differentiation induced by transforming growth factor-beta family members. PMID- 9417114 TI - The inhibition of fibroblast growth factor-2 export by cardenolides implies a novel function for the catalytic subunit of Na+,K+-ATPase. AB - Basic fibroblast growth factor (FGF-2) is one of a select group of proteins that can exit cells through an alternate, endoplasmic reticulum/Golgi apparatus independent exocytic pathway. This alternate pathway has been termed protein export. In an attempt to better understand this process, we have identified a family of related compounds, "cardenolides," that inhibit FGF-2 export. The cardenolides inhibit FGF-2 export in a time and concentration dependent fashion. Inhibition of FGF-2 export is specific in that the cardenolides have no effect on conventional protein secretion as measured by their inability to block release of the secreted protein human chorionic gonadotropin-alpha. Because cardenolides are known to inhibit ion transport activity mediated by Na+,K+-ATPase, we investigated whether there are functional interactions between FGF-2 and their only known molecular target: the alpha-subunit of Na+, K+-ATPase. Export of FGF-2 from COS-1 cells is selectively inhibited when co-transfected with expression vectors encoding the alpha-subunit and FGF-2. Moreover, antibodies to the alpha subunit specifically co-immunoprecipitate FGF-2 along with the alpha-subunit while conversely, antibodies to FGF-2 specifically co-immunoprecipitate the alpha subunit along with FGF-2. Finally, the ion transporting activities of the Na+,K+ ATPase can be uncoupled from protein export. Varying the external concentration of K+ has little effect on export of FGF-2. Taken together, these data: 1) identify a novel activity for cardenolides; 2) suggest a previously unknown role for the alpha-subunit of Na+, K+-ATPase in FGF-2 export; and 3) raise the possibility that the alpha-subunit itself may be an integral component of this alternate exocytic pathway mediating translocation of cytosolic FGF-2 to the cell surface. PMID- 9417115 TI - Coordinate transactivation of the interleukin-2 CD28 response element by c-Rel and ATF-1/CREB2. AB - The interleukin-2 CD28 response element (CD28RE) acts as a composite enhancer, in conjunction with a 3'-12-O-tetradecanoylphorbol-13-acetate response element (TRE) like element, to confer CD28 receptor-dependent inducibility to the interleukin-2 promoter in T-cells. When inserted as a single copy upstream of a basal promoter, this composite enhancer, termed the CD28RE-TRE, is both highly active and CD28 inducible in transactivation assays. A multicomponent nuclear protein complex that binds the CD28RE-TRE was isolated by DNA affinity chromatography from nuclear extracts of mitogen- and CD28 receptor-costimulated human T-cells. Immunological and biochemical analyses of this complex reveal the presence of c Rel, ATF-1, and CREB2 as major DNA-binding components. Coexpression of c-Rel in combination with ATF-1, CREB2, or ATF-1/CREB2 leads to synergistic transactivation of a CD28RE-TRE reporter plasmid in quiescent Jurkat T-cells. Furthermore, CD28-dependent transactivation of the CD28RE-TRE is specifically inhibited by cAMP response element-binding protein (CREB) dominant-negative expression vectors. Moreover, mutant promoter constructs in which the internal 5' CD28RE and 3'-TRE-like sequences have been topologically positioned 180 degrees out of phase with one another show loss of mitogen- and CD28-dependent inducibility. Finally, the addition of the CREB-binding transcriptional coactivator p300 leads to a dramatic CREB-dependent increase in both mitogen- and CD28-mediated transactivation of the CD28RE-TRE. These findings demonstrate that full physiological responsiveness to CD28 receptor stimulation in T-cells is dependent on topologically linked sequences within the CD28RE-TRE composite enhancer and provide strong support of a direct role for the CREB family of transcription factors and p300/CREB-binding protein coactivator proteins in cytokine gene induction during T-cell activation. PMID- 9417116 TI - A binding site for nuclear receptors is required for the differential expression of the aldolase A fast-twitch muscle promoter in body and head muscles. AB - In hind limb muscles, the aldolase A muscle-specific promoter is specifically expressed in glycolytic fast-twitch fibers. Here, we show that in addition, it is expressed at higher levels in trunk and limb muscles than in neck and head muscles independent of their fiber-type content. We have identified by analysis of transgenic mice a DNA element that is required for this differential expression and, to a lesser extent, for fiber-type specificity. We show that members of the nuclear receptor superfamily bind this element in skeletal muscle nuclear extracts. Interestingly, in gel mobility shift assays, different complexes were formed with this sequence in tongue nuclear extracts compared with limb or trunk muscle nuclear extracts. Therefore, binding of distinct nuclear receptors to a single regulatory sequence appears to be associated with the location-dependent expression of the aldolase A muscle-specific promoter. PMID- 9417117 TI - Co- and posttranslational translocation mechanisms direct cystic fibrosis transmembrane conductance regulator N terminus transmembrane assembly. AB - Transmembrane topology of most eukaryotic polytopic proteins is established cotranslationally at the endoplasmic reticulum membrane through the action of alternating signal and stop transfer sequences. Here we demonstrate that the cystic fibrosis transmembrane conductance regulator (CFTR) achieves its N terminus topology through a variation of this mechanism that involves both co- and posttranslational translocation events. Using a series of defined chimeric and truncated proteins expressed in a reticulocyte lysate system, we have identified two topogenic determinants encoded within the first (TM1) and second (TM2) membrane-spanning segments of CFTR. Each sequence independently (i) directed endoplasmic reticulum targeting, (ii) translocated appropriate flanking residues, and (iii) achieved its proper membrane-spanning orientation. Signal sequence activity of TM1, however, was inefficient due to the presence of two charged residues, Glu92 and Lys95, located within its hydrophobic core. As a result, TM1 was able to direct correct topology for less than half of nascent CFTR chains. In contrast to TM1, TM2 signal sequence activity was both efficient and specific. Even in the absence of a functional TM1 signal sequence, TM2 was able to direct CFTR N terminus topology through a ribosome-dependent posttranslational mechanism. Mutating charged residues Glu92 and Lys95 to alanine improved TM1 signal sequence activity as well as the ability of TM1 to independently direct CFTR N terminus topology. Thus, a single functional signal sequence in either the first or second TM segment was sufficient for directing proper CFTR topology. These results identify two distinct and redundant translocation pathways for CFTR N terminus transmembrane assembly and support a model in which TM2 functions to ensure correct topology of CFTR chains that fail to translocate via TM1. This novel arrangement of topogenic information provides an alternative to conventional cotranslational pathways of polytopic protein biogenesis. PMID- 9417118 TI - SH2- and SH3-mediated interactions between focal adhesion kinase and Src. AB - Intramolecular SH2 and SH3 interactions mediate enzymatic repression of the Src kinases. One mechanism of activation is disruption of these interactions by the formation of higher affinity SH2 and SH3 interactions with specific ligands. We show that a consensus Src SH3-binding site residing upstream of the Src SH2 binding site in FAK can function as a ligand for the Src SH3 domain. Surface plasmon resonance experiments indicate that a FAK peptide containing both the Src SH2- and SH3-binding sites exhibits increased affinity for Src. Furthermore, the presence of both sites in vitro more potently activates c-Src. A FAK mutant (FAKPro-2) with substitutions destroying the SH3-binding site shows reduced binding to Src in vivo. This mutation also reduces Src-dependent tyrosine phosphorylation on the mutant itself and downstream substrates, such as paxillin. These observations suggest that an SH3-mediated interaction between Src-like kinases and FAK may be important for complex formation and downstream signaling in vivo. PMID- 9417119 TI - Activation and processing of non-anchored yapsin 1 (Yap3p). AB - A C-terminally truncated form of yapsin 1 (yeast aspartic protease 3), the first member of the novel sub-class of aspartic proteases with specificity for basic residues (designated the Yapsins), was overexpressed and purified to apparent homogeneity, yielding approximately 1 microg of yapsin 1/g of wet yeast. N terminal amino acid analysis of the purified protein confirmed that the propeptide was absent and that the mature enzyme began at Ala68. The mature enzyme was shown to be composed of approximately equimolar amounts of two subunits, designated alpha and beta, that were associated to each other by a disulfide bond. C-terminally truncated proyapsin 1 was also expressed in the baculovirus/Sf9 insect cell expression system and secreted as a zymogen that could be activated upon incubation at an acidic pH with an optimum at approximately 4.0. When expressed without its pro-region, it was localized intracellularly and lacked activity, indicating that the pro-region was required for the correct folding of the enzyme. The activation of proyapsin 1 in vitro exhibited linear kinetics and generated an intermediate form of yapsin 1 or pseudo-yapsin 1. PMID- 9417120 TI - Discrimination between RelA and RelB transcriptional regulation by a dominant negative mutant of IkappaBalpha. AB - RelA and RelB belong to the nuclear factor-kappaB (NF-kappaB-Rel) transcription factor family. Both proteins are structurally and functionally related, but their intracellular and tissue distributions are different. In resting cells, RelB is found mostly in the nucleus, whereas RelA is sequestered in the cytosol by protein inhibitors, among which IkappaBalpha is the dominant form in lymphocytes. Upon cellular activation IkappaBalpha is proteolyzed, allowing RelA dimers to enter the nucleus and activate target genes. To study the selectivity of gene regulation by RelA and RelB, we generated T cell lines stably expressing a dominant negative mutant of IkappaBalpha. We show that selective inhibition of RelA-NF-kappaB decreased induction of NFKB1, interleukin-2, and interleukin 2Ralpha genes but not c-myc. Transcription driven by the IkappaBalpha promoter was blocked by the transgenic IkappaBalpha; however, wild type IkappaBalpha was expressed in the transgenic cell clones but with much slower kinetics than that in control cells. Wild type IkappaBalpha expression was concomitant with RelB up regulation, suggesting that RelB could be involved in transcription of IkappaBalpha through binding to an alternative site. These results indicate that RelB and RelA have both distinct and overlapping effects on gene expression. PMID- 9417121 TI - Agrin is a high-affinity binding protein of dystroglycan in non-muscle tissue. AB - Agrin is a basement membrane-associated proteoglycan that induces the formation of postsynaptic specializations at the neuromuscular junction. This activity is modulated by alternative splicing and is thought to be mediated by receptors expressed in muscle fibers. An isoform of agrin that does not induce postsynaptic specializations binds with high affinity to dystroglycan, a component of the dystrophin-glycoprotein complex. Transcripts encoding this agrin isoform are expressed in a variety of non-muscle tissues. Here, we analyzed the tissue distribution of agrin and dystroglycan on the protein level and determined their binding affinities. We found that agrin is most abundant in lung, kidney, and brain. Only a little agrin was detected in skeletal muscle, and no agrin was found in liver. Dystroglycan was highly expressed in all tissues examined except in liver. In a solid-phase radioligand binding assay, agrin bound to dystroglycan from lung, kidney, and skeletal muscle with a dissociation constant between 1.8 and 2.2 nM, while the affinity to brain-derived dystroglycan was 4.6 nM. In adult kidney and lung, agrin co-purified and co-immunoprecipitated with dystroglycan, and both molecules were co-localized in embryonic tissue. These data show that the agrin isoform expressed in non-muscle tissue is a high-affinity binding partner of dystroglycan and they suggest that this interaction, like that between laminin and dystroglycan, may be important for the mechanical integrity of the tissue. PMID- 9417122 TI - Secretory phospholipase A2 activates the cascade of mitogen-activated protein kinases and cytosolic phospholipase A2 in the human astrocytoma cell line 1321N1. AB - The biological effects of type IIA 14-kDa phospholipase A2 (sPLA2) on 1321N1 astrocytoma cells were studied. sPLA2 induced a release of [3H]arachidonic acid ([3H]AA) similar to that elicited by lysophosphatidic acid (LPA), a messenger acting via a G-protein-coupled receptor and a product of sPLA2 on lipid microvesicles. In contrast, no release of [1-14C]oleate could be detected in cells labeled with this fatty acid. As these findings pointed to a selective mechanism of [3H]AA release, it was hypothesized that sPLA2 could act by a signaling mechanism involving the activation of cytosolic PLA2 (cPLA2), i.e. the type of PLA2 involved in the release of [3H]AA elicited by agonists. In keeping with this view, stimulation of 1321N1 cells with sPLA2 elicited the decrease in electrophoretic mobility that is characteristic of the phosphorylation of cPLA2, as well as activation of p42 mitogen-activated protein (MAP) kinase, c-Jun kinase, and p38 MAP kinase. Incubation with sPLA2 of quiescent 1321N1 cells elicited a mitogenic response as judged from an increased incorporation of [3H]thymidine. Attempts to correlate the effect of extracellular PLA2 with the generation of LPA were negative. Incubation with pertussis toxin prior to the addition of either sPLA2 or LPA only showed abrogation of the response to LPA, thus suggesting the involvement of pertussis-sensitive Gi-proteins in the case of LPA. Treatments with inhibitors of the catalytic effect of sPLA2 such as p bromophenacyl bromide and dithiothreitol did not prevent the effect on cPLA2 activation. In contrast, preincubation of 1321N1 cells with the antagonist of the sPLA2 receptor p-aminophenyl-alpha-D-mannopyranoside-bovine serum albumin, blocked cPLA2 activation with a EC50 similar to that described for the inhibition of binding of sPLA2 to its receptor. Moreover, treatment of 1321N1 cells with the MAP kinase kinase inhibitor PD-98059 inhibited the activation of both cPLA2 and p42 MAP kinase produced by sPLA2. In summary, these data indicate the existence in astrocytoma cells of a signaling pathway triggered by engagement of a sPLA2 binding structure, that produces the release of [3H]AA by activating the MAP kinase cascade and cPLA2, and leads to a mitogenic response after longer periods of incubation. PMID- 9417123 TI - Involvement of Rabphilin3 in endocytosis through interaction with Rabaptin5. AB - Rabphilin3 and rabaptin5 are downstream target molecules of the Rab3 and -5 subfamily small G proteins that are implicated in exocytosis and endocytosis, respectively. We examined here the physical and functional relationship between the Rab3-rabphilin3 and Rab5-rabaptin5 systems. Rabphilin3 interacted with rabaptin5 at the N-terminal region (amino acids 1-280), which GTP-Rab3A interacted with. The interaction of rabphilin3 with rabaptin5 was inhibited by guanosine 5'-(3-O-thio)triphosphate-Rab3A. Overexpression of the N-terminal fragment of rabphilin3 (amino acids 1-280) inhibited the receptor-mediated endocytosis of transferrin, and this inhibition was overcome by co-transfection with a dominant active mutant of Rab3A or rabaptin5 in PC12 and HeLa cells. These results suggest that rabphilin3, free of GTP-Rab3A, regulates endocytosis through interaction with rabaptin5 after rabphilin3 complexed with GTP-Rab3A regulates exocytosis. PMID- 9417124 TI - Stromelysin-3 is induced in tumor/stroma cocultures and inactivated via a tumor specific and basic fibroblast growth factor-dependent mechanism. AB - Stromelysin-3 (STR-3) is a recently characterized matrix metalloproteinase (MMP) with a unique pattern of expression and substrate specificity. Unlike other MMPs, STR-3 is consistently and dramatically overexpressed by multiple epithelial malignancies, including carcinomas of the breast, lung, colon, head and neck, and skin. Recent studies suggest that STR-3 promotes the local establishment of epithelial malignancies, contributing to tumor cell survival and implantation in host tissues; however, STR-3's mechanism of action remains undefined. STR-3 is a stromal cell product, prompting speculation that infiltrating stromal cells secrete STR-3 in response to tumor-derived factors. To explore this possibility, we developed a tumor/"stroma" coculture assay in which non-small cell lung cancer (NSCLC) cell lines were grown on confluent monolayers of normal pulmonary fibroblasts. In these tumor/stroma cocultures, NSCLCs stimulate normal pulmonary fibroblasts to secrete STR-3 and release extracellular basic fibroblast growth factor. Thereafter, STR-3 is processed at a unique internal sequence via a basic fibroblast growth factor- and MMP-dependent mechanism to a previously unidentified 35-kDa protein that lacks enzymatic activity. 35-kDa STR-3 is the most abundant STR-3 protein in tumor/stroma cocultures and is only detected when normal pulmonary fibroblasts are cultured with malignant bronchial epithelial cells. Therefore, the tumor-specific processing of STR-3 to the 35-kDa protein is likely to be an important regulatory mechanism. PMID- 9417125 TI - Negative autoregulation of the organizer-specific homeobox gene goosecoid. AB - The homeobox gene goosecoid has been implicated to play a central role in the Spemann organizer tissue of the vertebrate embryo. Misexpression of goosecoid on the ventral side of a Xenopus laevis gastrula embryo was shown to result in a partial duplication of the primary body axis, reminiscent of the Spemann organizer graft. Normal embryonic development thus requires tight temporal and spatial control of genes instrumental for organizer function. In the present study we investigated the transcriptional control of goosecoid gene expression. Sequence analysis of the mouse and human promoter region revealed the presence of two palindromic binding elements for homeobox genes of the prd type to which goosecoid belongs. We show that Goosecoid protein can bind to these sites in vitro. By using reporter gene constructs of the human and mouse promoter, we demonstrate that Goosecoid can act as a repressor of its own promoter activity in transient co-transfection experiments in mouse P19 cells and in Xenopus embryos. Autorepression depends on the presence of the homeodomain and is mediated through the prd element more proximal to the transcriptional start site. Our results suggest a role for goosecoid in restricting organizer activity in the vertebrate gastrula embryo. PMID- 9417126 TI - The G protein beta5 subunit interacts selectively with the Gq alpha subunit. AB - The diversity in the heterotrimeric G protein alpha, beta, and gamma subunits may allow selective protein-protein interactions and provide specificity for signaling pathways. We examined the ability of five alpha subunits (alphai1, alphai2, alphao, alphas, and alphaq) to associate with three beta subunits (beta1, beta2, and beta5) dimerized to a gamma2 subunit containing an amino terminal hexahistidine-FLAG affinity tag (gamma2HF). Sf9 insect cells were used to overexpress the recombinant proteins. The hexahistidine-FLAG sequence does not hinder the function of the beta1gamma2HF dimer as it can be specifically eluted from an alphai1-agarose column with GDP and AlF4-, and purified beta1gamma2HF dimer stimulates type II adenylyl cyclase. The beta1gamma2HF and beta2gamma2HF dimers immobilized on an anti-FLAG affinity column bound all five alpha subunits tested, whereas the beta5gamma2HF dimer bound only alphaq. The ability of other alpha subunits to compete with the alphaq subunit for binding to the beta5gamma2HF dimer was tested. Addition of increasing amounts of purified, recombinant alphai1 to the alphaq in a Sf9 cell extract did not decrease the amount of alphaq bound to the beta5gamma2HF column. When G proteins in an extract of brain membranes were activated with GDP and AlF4- and deactivated in the presence of equal amounts of the beta1gamma2HF or beta5gamma2HF dimers, only alphaq bound to the beta5gamma2HF dimer. The alphaq-beta5gamma2HF interaction on the column was functional as GDP, and AlF4- specifically eluted alphaq from the column. These results indicate that although the beta1 and beta2 subunits interact with alpha subunits from the alphai, alphas, and alphaq families, the structurally divergent beta5 subunit only interacts with alphaq. PMID- 9417127 TI - Lipoarabinomannan of Mycobacterium tuberculosis promotes protein tyrosine dephosphorylation and inhibition of mitogen-activated protein kinase in human mononuclear phagocytes. Role of the Src homology 2 containing tyrosine phosphatase 1. AB - Lipoarabinomannan (LAM) is a putative virulence factor of Mycobacterium tuberculosis that inhibits monocyte functions, and this may involve antagonism of cell signaling pathways. The effects of LAM on protein tyrosine phosphorylation in cells of the human monocytic cell line THP-1 were examined. LAM promoted tyrosine dephosphorylation of multiple cell proteins and attenuated phorbol 12 myristate 13-acetate-induced activation of mitogen-activated protein kinase. To examine whether these effects of LAM could be related to activation of a phosphatase, fractions from LAM-treated cells were analyzed for dephosphorylation of para-nitrophenol phosphate. The data show that LAM induced increased phosphatase activity associated with the membrane fraction. The Src homology 2 containing tyrosine phosphatase 1 (SHP-1) is important for signal termination and was examined as a potential target of LAM. Exposure of cells to LAM brought about (i) an increase in tyrosine phosphorylation of SHP-1, and (ii) translocation of the phosphatase to the membrane. Phosphatase assay of SHP-1 immunoprecipitated from LAM-treated cells, using phosphorylated mitogen-activated protein kinase as substrate, indicated that LAM promoted increased activity of SHP-1 in vivo. LAM also activated SHP-1 directly in vitro. Exposure of cells to LAM also attenuated the expression of tumor necrosis factor-alpha, interleukin-12, and major histocompatibility class II molecules. These results suggest that one mechanism by which LAM deactivates monocytes involves activation of SHP-1. PMID- 9417128 TI - Activation of the leu-500 promoter by a reversed polarity tetA gene. Response to global plasmid supercoiling. AB - The leu-500 promoter is inactivated by a mutation in the -10 region but can be activated in topA Escherichia coli and Salmonella strains. We have found that the tetA gene plays a vital role in the topA-dependent activation of a plasmid-borne leu-500 promoter. In previous studies, the leu-500 promoter and tetA gene have been arranged divergently. In this study we have reversed the polarity of the tetA gene, thus locating the leu-500 promoter at the 3' end of tetA. Despite being formally located in the downstream region of tetA, the leu-500 promoter is equally well activated in a topA strain in this environment, even though it is 1.6 kilobase pairs away from the promoter of the reversed tetA gene. Activation of the leu-500 promoter depends on transcription and translation of tetA but is largely insensitive to the function of other transcription units on the plasmid. These results require a change in viewpoint of the role of tetA, from local to global supercoiling. We conclude that transcription of the tetA gene is the main generator of transcription-induced supercoiling that activates the leu-500 promoter. Unbalanced relaxation of this supercoiling leads to a net increase in the negative linking difference of the plasmid globally, and there is a linear correlation between the change in global plasmid topology and the activation of the leu-500 promoter. Thus the leu-500 promoter appears to respond to the negative supercoiling of the plasmid overall. PMID- 9417130 TI - Editorial PMID- 9417129 TI - The bovine calpastatin gene promoter and a new N-terminal region of the protein are targets for cAMP-dependent protein kinase activity. AB - To investigate the regulation of calpastatin gene expression, we isolated bovine heart calpastatin cDNAs and 5'-regions of the calpastatin gene. Analysis of 5' cDNA sequence identified a new translation initiation site that is in frame and 204 nucleotides upstream of the previously designated start site. Conceptual translation from this upstream AUG produces a protein containing 68 additional N terminal amino acids. This "XL" region contains three potential PKA phosphorylation sites but shares no homology with other regions of calpastatin or with any known protein. Immunoblot studies demonstrated that heart and liver contain a calpastatin protein of 145 kDa on SDS-polyacrylamide gel electrophoresis that comigrates with full-length bacterially expressed calpastatin and calpastatin produced by coupled in vitro transcription translation from the upstream AUG. An antibody raised against the XL region recognized the 145-kDa band, demonstrating that the upstream AUG is utilized and that the 145-kDa band represents full-length calpastatin in vivo. Transient transfection assays demonstrated that sequence within 272 nucleotides upstream of transcription initiation of the calpastatin gene is sufficient to direct moderate level transcription. Promoter sequences further upstream act to inhibit or stimulate transcriptional activity. Exposure of transfected cells to dibutyryl cAMP resulted in a 7-20-fold increase in promoter activity for constructs containing at least 272 nucleotides of upstream promoter sequence. Deletion analysis indicates that at least one cAMP-responsive element resides within 102 nucleotides of transcription initiation. PMID- 9417131 TI - The calculus of rod phototransduction. PMID- 9417132 TI - Kinetics of recovery of the dark-adapted salamander rod photoresponse. AB - The kinetics of the dark-adapted salamander rod photocurrent response to flashes producing from 10 to 10(5) photoisomerizations (Phi) were investigated in normal Ringer's solution, and in a choline solution that clamps calcium near its resting level. For saturating intensities ranging from approximately 10(2) to 10(4) Phi, the recovery phases of the responses in choline were nearly invariant in form. Responses in Ringer's were similarly invariant for saturating intensities from approximately 10(3) to 10(4) Phi. In both solutions, recoveries to flashes in these intensity ranges translated on the time axis a constant amount (tauc) per e fold increment in flash intensity, and exhibited exponentially decaying "tail phases" with time constant tauc. The difference in recovery half-times for responses in choline and Ringer's to the same saturating flash was 5-7 s. Above approximately 10(4) Phi, recoveries in both solutions were systematically slower, and translation invariance broke down. Theoretical analysis of the translation invariant responses established that tauc must represent the time constant of inactivation of the disc-associated cascade intermediate (R*, G*, or PDE*) having the longest lifetime, and that the cGMP hydrolysis and cGMP-channel activation reactions are such as to conserve this time constant. Theoretical analysis also demonstrated that the 5-7-s shift in recovery half-times between responses in Ringer's and in choline is largely (4-6 s) accounted for by the calcium-dependent activation of guanylyl cyclase, with the residual (1-2 s) likely caused by an effect of calcium on an intermediate with a nondominant time constant. Analytical expressions for the dim-flash response in calcium clamp and Ringer's are derived, and it is shown that the difference in the responses under the two conditions can be accounted for quantitatively by cyclase activation. Application of these expressions yields an estimate of the calcium buffering capacity of the rod at rest of approximately 20, much lower than previous estimates. PMID- 9417133 TI - Onset of feedback reactions underlying vertebrate rod photoreceptor light adaptation. AB - Light adaptation in vertebrate photoreceptors is thought to be mediated through a number of biochemical feedback reactions that reduce the sensitivity of the photoreceptor and accelerate the kinetics of the photoresponse. Ca2+ plays a major role in this process by regulating several components of the phototransduction cascade. Guanylate cyclase and rhodopsin kinase are suggested to be the major sites regulated by Ca2+. Recently, it was proposed that cGMP may be another messenger of light adaptation since it is able to regulate the rate of transducin GTPase and thus the lifetime of activated cGMP phosphodiesterase. Here we report measurements of the rates at which the changes in Ca2+ and cGMP are followed by the changes in the rates of corresponding enzymatic reactions in frog rod outer segments. Our data indicate that there is a temporal hierarchy among reactions that underlie light adaptation. Guanylate cyclase activity and rhodopsin phosphorylation respond to changes in Ca2+ very rapidly, on a subsecond time scale. This enables them to accelerate the falling phase of the flash response and to modulate flash sensitivity during continuous illumination. To the contrary, the acceleration of transducin GTPase, even after significant reduction in cGMP, occurs over several tens of seconds. It is substantially delayed by the slow dissociation of cGMP from the noncatalytic sites for cGMP binding located on cGMP phosphodiesterase. Therefore, cGMP-dependent regulation of transducin GTPase is likely to occur only during prolonged bright illumination. PMID- 9417134 TI - Bleached pigment produces a maintained decrease in outer segment Ca2+ in salamander rods. AB - A spot confocal microscope based on an argon ion laser was used to make measurements of cytoplasmic calcium concentration (Ca2+i) from the outer segment of an isolated rod loaded with the fluorescent calcium indicator fluo-3 during simultaneous suction pipette recording of the photoresponse. The decline in fluo 3 fluorescence from a rod exposed to saturating illumination was best fitted by two exponentials of approximately equal amplitude with time constants of 260 and 2,200 ms. Calibration of fluo-3 fluorescence in situ yielded Ca2+i estimates of 670 +/- 250 nM in a dark-adapted rod and 30 +/- 10 nM during response saturation after exposure to bright light (mean +/- SD). The resting level of Ca2+i was significantly reduced after bleaching by the laser spot, peak fluo-3 fluorescence falling to 56 +/- 5% (SEM, n = 9) of its value in the dark-adapted rod. Regeneration of the photopigment with exogenous 11-cis-retinal restored peak fluo 3 fluorescence to a value not significantly different from that originally measured in darkness, indicating restoration of the dark-adapted level of Ca2+i. These results are consistent with the notion that sustained activation of the transduction cascade by bleached pigment produces a sustained decrease in rod outer segment Ca2+i, which may be responsible for the bleach-induced adaptation of the kinetics and sensitivity of the photoresponse. PMID- 9417135 TI - Voltage-dependent membrane displacements measured by atomic force microscopy. AB - Cells use polar molecules in the membrane to sense changes in the transmembrane potential. The opening of voltage-gated ion channels and membrane bending due to the inverse flexoelectric effect are two examples of such electromechanical coupling. We have looked for membrane motions in an electric field using atomic (or scanning) force microscopy (AFM) with the intent of studying voltage dependent conformational changes of ion channels. Voltage-clamped HEK293 cells were either untransfected controls or transfected with Shaker K+ channels. Using a +/- 10-mV peak-peak AC carrier stimulus, untransfected cells moved 0.5-15 nm normal to the plane of the membrane. These movements tracked the voltage at frequencies >1 kHz with a phase lead of 60-120 degrees, as expected of a displacement current. The movement was outward with depolarization, but the holding potential only weakly influenced the amplitude of the movement. In contrast, cells transfected with a noninactivating mutant of Shaker K+channels showed similar movements, but these were sensitive to the holding potential; decreasing with depolarization between -80 and 0 mV. Searching for artifactual origins of these movements, we used open or sealed pipettes and AFM cantilever placements just above the cells. These results were negative, suggesting that the observed movements were produced by the cell membrane rather than by movement of the patch pipette, or by acoustic or electrical interactions of the membrane with the AFM tip. In control cells, the electrical motor may arise from the flexoelectric effect, where changes in potential induce changes in curvature. In transfected cells, it appears that channel-specific movements also occurred. These experiments demonstrate that the AFM may be able to exploit voltage dependent movements as a source of contrast for imaging membrane components. The electrically induced motility will cause twitching during action potentials, and may have physiological consequences. PMID- 9417136 TI - Kinetic analysis of block of open sodium channels by a peptide containing the isoleucine, phenylalanine, and methionine (IFM) motif from the inactivation gate. AB - We analyzed the kinetics of interaction between the peptide KIFMK, containing the isoleucine, phen-ylalanine, and methionine (IFM) motif from the inactivation gate, and the brain type IIA sodium channels with a mutation that disrupts inactivation (F1489Q). The on-rate constant was concentration dependent, consistent with a bimolecular reaction with open sodium channels, while the off rates were unaffected by changes in the KIFMK concentration. The apparent Kd was approximately 33 microM at 0 mV. The on rates were voltage dependent, supporting the hypothesis that one or both of the charges in KIFMK enter the membrane electric field. The voltage dependence of block was consistent with the equivalent movement of approximately 0.6 electronic charges across the membrane. In contrast, the off rates were voltage independent. The results are consistent with the hypothesis that the KIFMK peptide enters the pore of the open sodium channel from the intracellular side and blocks it. PMID- 9417137 TI - Slow inactivation does not affect movement of the fast inactivation gate in voltage-gated Na+ channels. AB - Voltage-gated Na+ channels exhibit two forms of inactivation, one form (fast inactivation) takes effect on the order of milliseconds and the other (slow inactivation) on the order of seconds to minutes. While previous studies have suggested that fast and slow inactivation are structurally independent gating processes, little is known about the relationship between the two. In this study, we probed this relationship by examining the effects of slow inactivation on a conformational marker for fast inactivation, the accessibility of a site on the Na+ channel III-IV linker that is believed to form a part of the fast inactivation particle. When cysteine was substituted for phenylalanine at position 1304 in the rat skeletal muscle sodium channel (microl), application of [2-(trimethylammonium)ethyl]methanethiosulfonate (MTS-ET) to the cytoplasmic face of inside-out patches from Xenopus oocytes injected with F1304C RNA dramatically disrupted fast inactivation and displayed voltage-dependent reaction kinetics that closely paralleled the steady state availability (hinfinity) curve. Based on this observation, the accessibility of cys1304 was used as a conformational marker to probe the position of the fast inactivation gate during the development of and the recovery from slow inactivation. We found that burial of cys1304 is not altered by the onset of slow inactivation, and that recovery of accessibility of cys1304 is not slowed after long (2-10 s) depolarizations. These results suggest that (a) fast and slow inactivation are structurally distinct processes that are not tightly coupled, (b) fast and slow inactivation are not mutually exclusive processes (i.e., sodium channels may be fast- and slow-inactivated simultaneously), and (c) after long depolarizations, recovery from fast inactivation precedes recovery from slow inactivation. PMID- 9417138 TI - Pacemaker synchronization of electrically coupled rabbit sinoatrial node cells. AB - The effects of intercellular coupling conductance on the activity of two electrically coupled isolated rabbit sinoatrial nodal cells were investigated. A computer-controlled version of the "coupling clamp" technique was used in which isolated sinoatrial nodal cells, not physically in contact with each other, were electrically coupled at various values of ohmic coupling conductance, mimicking the effects of mutual interaction by electrical coupling through gap junctional channels. We demonstrate the existence of four types of electrical behavior of coupled spontaneously active cells. As the coupling conductance is progressively increased, the cells exhibit: (a) independent pacemaking at low coupling conductances, (b) complex dynamics of activity with mutual interactions, (c) entrainment of action potential frequency at a 1:1 ratio with different action potential waveforms, and (d) entrainment of action potentials at the same frequency of activation and virtually identical action potential waveforms. The critical value of coupling conductance required for 1:1 frequency entrainment was <0.5 nS in each of the five cell pairs studied. The common interbeat interval at a relatively high coupling conductance (10 nS), which is sufficient to produce entrainment of frequency and also identical action potential waveforms, is determined most by the intrinsically faster pacemaker cell and it can be predicted from the diastolic depolarization times of both cells. Evidence is provided that, at low coupling conductances, mutual pacemaker synchronization results mainly from the phase-resetting effects of the action potential of one cell on the depolarization phase of the other. At high coupling conductances, the tonic, diastolic interactions become more important. PMID- 9417139 TI - Nitric oxide signaling mediates stimulation of L-type Ca2+ current elicited by withdrawal of acetylcholine in cat atrial myocytes. AB - A perforated-patch whole-cell recording method was used to determine whether nitric oxide signaling participates in acetylcholine (ACh)-induced regulation of basal L-type Ca2+ current (ICa,L) in cat atrial myocytes. Exposure to 1 microM ACh for 2 min inhibited basal ICa,L (-21 +/- 3%), and withdrawal of ACh elicited rebound stimulation of ICa,L above control (80 +/- 13%) (n = 23). Stimulation of ICa,L elicited by withdrawal of ACh (but not ACh-induced inhibition of ICa,L) was blocked by either 50 microM hemoglobin; 30 microM ODQ or 10 microM methylene blue, inhibitors of soluble guanylate cyclase; 10 microM W-7, a calmodulin inhibitor; or 10 microM L-NIO, an inhibitor of constitutive NO synthase (NOS). In cells incubated in 5 mM L-arginine, ACh-induced rebound stimulation of ICa,L was enhanced compared with control responses. Histochemical assay (NADPH diaphorase) indicated that atrial myocytes express constitutive NOS. NO-donor, spermine/NO (SP/NO), >1 microM stimulated basal ICa,L. SP/NO-induced stimulation of ICa,L was inhibited by 50 microM hemoglobin, 30 microM ODQ, or 5 microM H-89, an inhibitor of PKA, and was unchanged by 50 microM MnTBAP, a peroxynitrite scavenger. When ICa,L was prestimulated by 10 microM milrinone, an inhibitor of cGMP-inhibited phosphodiesterase (type III) activity, SP/NO failed to further increase ICa,L. In cells incubated in pertussis toxin (3.4 microg/ml for 6 h; 36 degrees C), ACh failed to affect ICa,L, but 100 microM SP/NO or 10 microM milrinone still increased basal ICa,L. These results indicate that in cat atrial myocytes NO signaling mediates stimulation of ICa,L elicited by withdrawal of ACh but not ACh induced inhibition of basal ICa,L. NO activates cGMP-induced inhibition of phosphodiesterase (type III) activity. Upon withdrawal of ACh, this mechanism allows cAMP to recover to levels above control, thereby stimulating ICa,L. Pertussis toxin-sensitive G-proteins couple M2 muscarinic receptors to NO signaling. NO-mediated stimulation of ICa, L elicited by withdrawal of ACh may be an important mechanism that rapidly restores cardiac pacemaker and contractile functions after cholinergic suppression of atrial activity. PMID- 9417140 TI - Protease modulation of the activity of the epithelial sodium channel expressed in Xenopus oocytes. AB - We have investigated the effect of extracellular proteases on the amiloride sensitive Na+ current (INa) in Xenopus oocytes expressing the three subunits alpha, beta, and gamma of the rat or Xenopus epithelial Na+ channel (ENaC). Low concentrations of trypsin (2 microg/ml) induced a large increase of INa within a few minutes, an effect that was fully prevented by soybean trypsin inhibitor, but not by amiloride. A similar effect was observed with chymotrypsin, but not with kallikrein. The trypsin-induced increase of INa was observed with Xenopus and rat ENaC, and was very large (approximately 20-fold) with the channel obtained by coexpression of the alpha subunit of Xenopus ENaC with the beta and gamma subunits of rat ENaC. The effect of trypsin was selective for ENaC, as shown by the absence of effect on the current due to expression of the K+ channel ROMK2. The effect of trypsin was not prevented by intracellular injection of EGTA nor by pretreatment with GTP-gammaS, suggesting that this effect was not mediated by G proteins. Measurement of the channel protein expression at the oocyte surface by antibody binding to a FLAG epitope showed that the effect of trypsin was not accompanied by an increase in the channel protein density, indicating that proteolysis modified the activity of the channel present at the oocyte surface rather than the cell surface expression. At the single channel level, in the cell attached mode, more active channels were observed in the patch when trypsin was present in the pipette, while no change in channel activity could be detected when trypsin was added to the bath solution around the patch pipette. We conclude that extracellular proteases are able to increase the open probability of the epithelial sodium channel by an effect that does not occur through activation of a G protein-coupled receptor, but rather through proteolysis of a protein that is either a constitutive part of the channel itself or closely associated with it. PMID- 9417144 TI - Children who prosper in unfavorable environments: the relationship to social capital. AB - OBJECTIVE: Social capital describes the benefits that are derived from personal social relationships (within families and communities) and social affiliations. This investigation examined the extent to which social capital is associated with positive developmental and behavioral outcomes in high-risk preschool children. DESIGN: A cross-sectional case-control analysis of young children "doing well" and "not doing well" at baseline in four coordinated longitudinal studies. PARTICIPANTS: A total of 667 2- to 5-year-old children (mean age, 4.4 years) and their maternal caregivers who are participating in the Longitudinal Studies of Child Abuse and Neglect Consortium. At recruitment, all children were characterized by unfavorable social or economic circumstances that contributed to the identification of the children as high risk. MEASURES: Social capital was defined as benefits that accrue from social relationships within communities and families. A social capital index was created by assigning one point to each of the following indicators: 1) two parents or parent-figures in the home; 2) social support of the maternal caregiver; 3) no more than two children in the family; 4) neighborhood support; and 5) regular church attendance. Outcomes were measured with the Child Behavior Checklist, a widely used measure of behavioral/emotional problems, and with the Battelle Developmental Inventory Screening Test, a standardized test that identifies developmental deficits. Children were classified as doing well if their scores on these instruments indicated neither behavioral nor developmental problems. RESULTS: Only 13% of the children were classified as doing well. The individual indicators that best discriminated between levels of child functioning were the most direct measures of social capital-church affiliation, perception of personal social support, and support within the neighborhood. The social capital index was strongly associated with child well-being, more so than any single indicator. The presence of any social capital indicator increased the odds of doing well by 29%; adding any two increased the odds of doing well by 66%. CONCLUSIONS: Our findings suggest that social capital may have an impact on children's well-being as early as the preschool years. In these years it seems to be the parents' social capital that confers benefits on their offspring, just as children benefit from their parents' financial and human capital. Social capital may be most crucial for families who have fewer financial and educational resources. Our findings suggest that those interested in the healthy development of children, particularly children most at risk for poor developmental outcomes, must search for new and creative ways of supporting interpersonal relationships and strengthening the communities in which families carry out the daily activities of their lives. PMID- 9417141 TI - Properties of an inwardly rectifying ATP-sensitive K+ channel in the basolateral membrane of renal proximal tubule. AB - The potassium conductance of the basolateral membrane (BLM) of proximal tubule cells is a critical regulator of transport since it is the major determinant of the negative cell membrane potential and is necessary for pump-leak coupling to the Na+,K+-ATPase pump. Despite this pivotal physiological role, the properties of this conductance have been incompletely characterized, in part due to difficulty gaining access to the BLM. We have investigated the properties of this BLM K+ conductance in dissociated, polarized Ambystoma proximal tubule cells. Nearly all seals made on Ambystoma cells contained inward rectifier K+ channels (gammaslope, in = 24.5 +/- 0.6 pS, gammachord, out = 3.7 +/- 0.4 pS). The rectification is mediated in part by internal Mg2+. The open probability of the channel increases modestly with hyperpolarization. The inward conducting properties are described by a saturating binding-unbinding model. The channel conducts Tl+ and K+, but there is no significant conductance for Na+, Rb+, Cs+, Li+, NH4+, or Cl-. The channel is inhibited by barium and the sulfonylurea agent glibenclamide, but not by tetraethylammonium. Channel rundown typically occurs in the absence of ATP, but cytosolic addition of 0. 2 mM ATP (or any hydrolyzable nucleoside triphosphate) sustains channel activity indefinitely. Phosphorylation processes alone fail to sustain channel activity. Higher doses of ATP (or other nucleoside triphosphates) reversibly inhibit the channel. The K+ channel opener diazoxide opens the channel in the presence of 0.2 mM ATP, but does not alleviate the inhibition of millimolar doses of ATP. We conclude that this K+ channel is the major ATP-sensitive basolateral K+ conductance in the proximal tubule. PMID- 9417143 TI - A comparative efficacy trial in Germany in infants who received either the Lederle/Takeda acellular pertussis component DTP (DTaP) vaccine, the Lederle whole-cell component DTP vaccine, or DT vaccine. AB - BACKGROUND: The goal of the trial was to determine the efficacy of a multicomponent acellular pertussis vaccine against Bordetella illnesses in comparison with a whole-cell product and DT. DESIGN: In a randomized, double blind fashion, 2- to 4-month-old infants received 4 doses of either DTP or DTaP vaccine at 3, 4.5, 6, and 15 to 18 months of age. The controls received 3 doses (3, 4.5, 15 to 18 months of age) of DT vaccine. The DTP vaccine was Lederle adsorbed vaccine (licensed in the United States) and DTaP was Lederle/Takeda adsorbed vaccine. Follow-up for vaccine efficacy started 2 weeks after the third dose (DTP/DTaP) and at the same age (6.5 months) in DT recipients. Reactogenicity of all doses of all three vaccines was documented by standardized parent diary cards. In addition, all subjects were monitored for respiratory illnesses and serious adverse events by biweekly phone calls. RESULTS: From May 1991 to January 1993, a total of 10 271 infants were enrolled: 8532 received either DTP or DTaP and 1739 received DT. Specific efficacy against B pertussis infections with cough >/=7 days duration was 83% (95% confidence interval [CI]: 76-88) and 72% (95% CI: 62-79) for DTP and DTaP, respectively; results for DTP and DTaP based on >/=21 days of cough with either paroxysms, whoop or posttussive vomiting (PWV) were 93% (95% CI: 89-96) and 83% (95% CI: 76-88), respectively. For DTaP vaccine, efficacy was higher after the fourth dose as compared with its efficacy after the third dose (78% vs 62% for cough >/=7 days and 85% vs 76% for cough >/=21 days with PWV). For DTP vaccine, efficacy was less varied after the third and fourth dose (78% vs 85% for cough >/=7 days and 93% vs 93% for cough >/=21 days with PWV). In contrast with DTP, the DTaP vaccine had some efficacy against B parapertussis infection (point estimate for cough >/=7 days: 31% [95% CI: -10-56]). All vaccines were generally well-tolerated. However, side reactions were significantly less after DTaP compared with DTP. CONCLUSIONS: Like other multicomponent acellular pertussis vaccines, the Lederle/Takeda DTaP vaccine demonstrated good efficacy against mild and typical pertussis due to B pertussis infections. Interestingly, it also may have some efficacy against B parapertussis. Based on the results of this trial, the vaccine was licensed in the United States in December 1996 for all 5 doses of the currently recommended immunization schedule in this country. PMID- 9417142 TI - Regulation of an inwardly rectifying ATP-sensitive K+ channel in the basolateral membrane of renal proximal tubule. AB - Functional coupling of Na+,K+-ATPase pump activity to a basolateral membrane (BLM) K+ conductance is crucial for sustaining transport in the proximal tubule. Apical sodium entry stimulates pump activity, lowering cytosolic [ATP], which in turn disinhibits ATP-sensitive K+ (KATP) channels. Opening of these KATP channels mediates hyperpolarization of the BLM that facilitates Na+ reabsorption and K+ recycling required for continued Na+,K+-ATPase pump turnover. Despite its physiological importance, little is known about the regulation of this channel. The present study focuses on the regulation of the BLM KATP channel by second messengers and protein kinases using membrane patches from dissociated, polarized Ambystoma proximal tubule cells. The channel is regulated by protein kinases A and C, but in opposing directions. The channel is activated by forskolin in cell attached (c/a) patches, and by PKA in inside-out (i/o) membrane patches. However, phosphorylation by PKA is not sufficient to prevent channel rundown. In contrast, the channel is inhibited by phorbol ester in c/a patches, and PKC decreases channel activity (nPo) in i/o patches. The channel is pH sensitive, and lowering cytosolic pH reduces nPo. Increasing intracellular [Ca2+] ([Ca2+]i) in c/a patches decreases nPo, and this effect is direct since [Ca2+]i inhibits nPo with a Ki of approximately 170 nM in i/o patches. Membrane stretch and hypotonic swelling do not significantly affect channel behavior, but the channel appears to be regulated by the actin cytoskeleton. Finally, the activity of this BLM KATP channel is coupled to transcellular transport. In c/a patches, maneuvers that inhibit turnover of the Na+,K+-ATPase pump reduce nPo, presumably due to a rise in intracellular [ATP], although the associated cell depolarization cannot be ruled out as the possible cause. Conversely, stimulation of transport (and thus pump turnover) leads to increases in nPo, presumably due to a fall in intracellular [ATP]. These results show that the inwardly rectifying KATP channel in the BLM of the proximal tubule is a key element in the feedback system that links cellular metabolism with transport activity. We conclude that coupling of this KATP channel to the activity of the Na+,K+-ATPase pump is a mechanism by which steady state NaCl reabsorption in the proximal tubule may be maintained. PMID- 9417145 TI - When should a child be in the hospital? A. Frederick North, Jr, MD, revisited. AB - OBJECTIVES: My objective was to revisit the issues and approaches raised in a seminal article published in Pediatrics in 1976 by A. Frederick North, Jr, entitled "When Should a Child Be in the Hospital?" Dr North proposed a set of nine criteria to guide the evaluation of appropriateness of admission to hospital. These were based on a core assertion that, "The need to hospitalize a child is dependent on the special services which the child requires rather than upon the diagnosis." This original work antedated more recent activities and publications in the area of appropriateness evaluation as applied to pediatrics (such as the Pediatric Appropriateness Evaluation Protocol), but is more context specific than later works in the field. METHODS: A review of the literature concerning temporal trends in hospital use for children in North America was undertaken. This was done to place some of the subsequent observations in a macrocontext of overall trends in hospital use. A review of the English language literature focusing on alternatives to hospitalization and contextual evolution affecting patterns of hospital care for children is presented. Factors influencing each of the nine admission criteria proposed by North are reviewed and discussed in turn. RESULTS: Overall rates of hospitalization declined by 46% and 41% in the United States and Canada, respectively, during the 1971 to 1993 interval. The relative composition by diagnostic category in two specific pediatric hospital settings also evolved substantially during the 2 decades. Many of North's specific criteria required extensive revision or updating to match the contextual realities of the 1990s. CONCLUSIONS: Although important overall shifts have occurred in the absolute levels and relative composition of pediatric hospitalization, Dr North's core assertion, relating to the need for specific services driving the need for hospitalization, has largely stood the test of time. PMID- 9417146 TI - Practice patterns in neonatal hyperbilirubinemia. AB - OBJECTIVE: To determine practice patterns of office-based pediatricians and neonatologists in the treatment of neonatal hyperbilirubinemia in healthy, term newborns during 1992, before the publication of the practice guideline for treatment of neonatal jaundice by the American Academy of Pediatrics (AAP). The survey was undertaken to inform the AAP's Subcommittee on Hyperbilirubinemia on current practices and to aid it in its preparation of the guidelines. It was also anticipated that this survey would serve as a basis for comparison for a second survey to be performed several years after the publication of the practice guidelines. METHODS: A self-administered questionnaire describing a single case of a jaundiced, breastfed 36-hour-old healthy, full-term infant with a total serum bilirubin concentration of 11.0 mg/dL (188 microM/L) was sent to a random sample of 600 office-based pediatricians and 606 neonatologists who were members of the AAP. The final response rate was 74%. Respondents were asked to answer questions regarding treatment of the case based on their actual practices. Ranges of total serum bilirubin concentration were provided as possible answers to questions on initiation of phototherapy and exchange transfusion, and interruption of breastfeeding. Respondents were also queried about frequency of serum bilirubin testing, locations of phototherapy administration, and factors influencing their therapeutic decisions. RESULTS: Four hundred forty-two office based pediatricians and 444 neonatologists completed the survey. There was a tendency for neonatologists to initiate both phototherapy and exchange transfusions at lower serum bilirubin concentrations than office-based general pediatricians. At a serum bilirubin of 13 to 19 mg/dL (222 to 325 microM/L), 54% of office-based pediatricians stated they would initiate phototherapy whereas 76% of neonatologists would do so. Forty percent of office-based practitioners said they would perform exchange transfusions at serum bilirubin levels of 20 to 25 mg/dL (342 to 428 microM/L), whereas 60% of neonatologists said they would. Only a small percentage of both office-based practitioners (13%) and neonatologists (16%) indicated they would interrupt breastfeeding at 8 to 13 mg/dL (137 to 222 microM/L); but with each incremental level of serum bilirubin, an increasing proportion of neonatologists would interrupt breastfeeding. Little correlation was found between treatment practices and demographic characteristics except for years in practice; physicians with the fewest years in practice (5 years or less) differed significantly from all other groups of physicians in initiating exchange transfusions at higher serum bilirubin concentrations. CONCLUSIONS: The results of this survey indicated a wide range of variation of opinion among both groups of physicians, most likely a reflection of the uncertainty and controversy surrounding these issues. The data may also reflect a possible wide range of "acceptable practice" as opposed to a narrow treatment standard. Office-based practitioners more closely approximated the new 1994 recommendations than neonatologists. PMID- 9417148 TI - Resident and family continuity in pediatric continuity clinic: nine years of observation. AB - OBJECTIVE: To assess resident, patient, and family continuity. BACKGROUND: Continuity clinic is the principal longitudinal primary care experience for pediatric residents. Although it has been a recommendation of the Residency Review Committee for pediatric training for more than 10 years and has been a requirement of the Accreditation Council of Graduate Medical Education since 1989, the extent to which continuity is achieved in this setting has not been reported. METHODS: Nine years (1984-1993) of residents' continuity clinic experience in a community hospital affiliate of a university training program were reviewed. Continuity was defined by recurring visits between the same patient/provider pair. The analysis from 57 different residents includes 48 intern (R1) years, 45 level two (R2) years, and 40 level three (R3) years; 32 of these residents completed all 3 years of training (3-year cohort) in the program during the study period. Observations included 89 952 visits by 11 009 patients in 7130 families. Continuity was determined for the resident, patient, and family. RESULTS: Residents saw an annual average of 93, 136, and 144 visits as R1s, R2s, and R3s. Residents saw 60% of their patients fewer than 3 times and nearly 40% only once. In the final year for those in the 3-year cohort, residents saw an average of 149 visits; 53% of the time these R3s had seen their patients once or twice over 3 years. Thirty percent of the patients never saw their primary care physician (PCP) and 72% of patients had fewer than 3 visits with their PCP. One quarter of the families never saw their continuity resident, and 62% saw their continuity resident fewer than 3 times. CONCLUSIONS: These data demonstrate a remarkable lack of both resident and patient continuity in the principal clinical activity affording longitudinal primary care experiences during residency training. If more continuity is essential for both primary care of patients and education in general pediatrics, change in the structure of continuity experience is required. PMID- 9417147 TI - Neonatal hospital lengths of stay, readmissions, and charges. AB - OBJECTIVE: To evaluate trends in length of hospital stay, hospital charges, and readmission rates of Wisconsin newborns from 1989 through 1994 in light of recent policies requiring earlier discharges after delivery of newborns. METHODOLOGY: Two data sources were used: 1) 1989-1994 Hospital Inpatient Discharge Data from the Wisconsin Office of Health Care Information, and 2) 1994 birth certificate and matched infant mortality data from the Wisconsin Center for Health Statistics. Average lengths of stay and average hospital (delivery and readmission) charges were calculated, and readmission rates were estimated for full-term, premature, and sick newborns. RESULTS: There were 368 955 full-term and 26 668 premature newborns in Wisconsin from 1989 through 1994. The average length of stay decreased by 24% in full-term newborns from 1989 through 1994, while average hospital (delivery and readmission) charges rose over 40% during the same period. Average length of stay for premature infants increased by 24% while their hospital delivery charges increased 214% during the study period. Readmission rates halved, yet charges per readmission doubled for full-term infants. More than twice as many full-term newborns were classified as sick in 1994 (43%) compared with 1989 (19%). CONCLUSIONS: Managed care efforts to control costs of neonatal care through earlier newborn discharge policies may have limited impact. Physicians or hospitals may be compensating for these policies by classifying more newborns as sick, thereby allowing for longer hospital stays to be reimbursed by the insurance carriers. Premature infants, <7% of the total births, account for half of all hospital delivery charges. Efforts to reduce premature births may have a greater impact on neonatal health care costs than efforts to discharge full-term newborns earlier. PMID- 9417149 TI - Human papillomavirus screening in pediatric victims of sexual abuse. AB - OBJECTIVE: To evaluate for the presence of subclinical human papillomavirus (HPV) in cases of suspected sexual abuse in children. DESIGN: Prospective data collection via interviews, physical examination, colposcopic examination, and tissue sampling by a surface swab technique. SETTING: A total of 40 pediatric patients ranging in age from 1 to 16 years who were referred to the Special Assessment and Management Clinic at Cardinal Glennon Children's Hospital, St Louis, MO, for probable or confirmed sexual abuse. INTERVENTIONS: In addition to colposcopic examination for physical signs of abuse, the patients were screened for evidence of sexually transmitted diseases, including syphilis, gonorrhea, and Chlamydia. At that time, surveillance sampling of the throat, vaginal introitus, and/or rectum by a simple, rapid surface swab technique was performed to detect the presence of HPV. MEASUREMENTS: Template DNA was extracted from cotton swabs and analyzed using polymerase chain reaction analysis. RESULTS: Human beta-globin sequences were detected in 58 (83%) of 70 specimens obtained from 40 patients, indicating successful processing had occurred. Using a consensus L1 primer-probe set capable of detecting multiple HPV genotypes, 2 (3%) of 58 samples from 2 (5%) of 40 patients were positive for HPV 16. None of the other 56 specimens yielded evidence of HPV. Appropriate positive and negative controls were included in each assay. CONCLUSIONS: Our results suggest that subclinical HPV infection is possible, but not commonly associated with sexual abuse in children from St Louis, MO. In this group of children without condyloma, HPV 16 was the only type identified. PMID- 9417150 TI - Sodium space and intravascular volume: dietary sodium effects in cystic fibrosis and healthy adolescent subjects. AB - OBJECTIVE: To assess the physiologic response to salt depletion in subjects with cystic fibrosis (CF) and control male adolescents for sodium balance, sodium space, and stimulation of the renin-angiotensin-aldosterone axis. DESIGN: Seven subjects with CF and six controls received a salt-replete (150 or 290 mmol NaCl per day) diet and then a salt-deplete (10 mmol NaCl per day) diet while in a clinical research center. RESULTS: Space maintenance: CF subjects responded to salt depletion with a greater weight loss than did controls (1.9 vs 0.8 kg) and a decrease in 24Na+ space, whereas controls maintained 24Na+ space. Paired (Na deplete/Na-replete) blood volumes decreased in subjects with CF, but not in controls. Renin-angiotensin-aldosterone axis stimulation: During salt repletion, subjects with CF had significantly higher aldosterone values than did controls in the afternoon, but not at 7:00 AM. During salt depletion, plasma renin activity and aldosterone increased significantly more in subjects with CF than in controls (renin, 35 vs 13 ng/mL/hour [9.7 vs 3.6 ng.L-1 s-1]; aldosterone: 181 vs 101 ng/dL [5021 vs 2802 pmol/L]). Furthermore, the angiotensin antagonist saralasin increased renin much more in subjects with CF (154 vs 36 ng/mL per hour [43 vs 10 ng.L-1 s-1]). Vasomotor functions: Mean arterial pressure was decreased in subjects with CF on both diets and decreased significantly more with low salt only in subjects with CF. During salt depletion, subjects with CF showed enhanced orthostatic tolerance (less heart rate increase with standing) compared with controls, thus obscuring their volume loss. The blood pressure response to an acute infusion of saralasin suggested that in salt-replete subjects with CF, but not in controls, angiotensin receptors were functional in maintaining vascular tone. During salt depletion, angiotensin was more important for maintenance of blood pressure in subjects with CF than in controls, because the saralasin induced drop in blood pressure was 20%, ie, close to shock levels, in subjects with CF, and only 6% in controls. CONCLUSION: The data suggest that patients with CF are so successful in compensating for volume depletion by vigorous activation of the renin-angiotensin system that salt depletion/dehydration cannot be recognized easily by routine clinical measurements, eg, capillary refill, serum sodium levels, or tachycardia. PMID- 9417151 TI - Moyamoya syndrome associated with congenital heart disease. AB - OBJECTIVE: To describe the association between moyamoya syndrome and congenital heart disease and to discuss its clinical implications. Study Design. Retrospective analysis of a case series from two institutions. RESULTS: Five patients with moyamoya syndrome and structural congenital heart disease were identified. Coarctation of the aorta was present in 3 patients, in association with a ventricular septal defect (1 patient), aortic and mitral valve stenoses (1 patient), and tetralogy of Fallot (1 patient). Tetralogy of Fallot and a large paramembranous ventricular septal defect were found in the other 2 patients. Four patients underwent surgical repair of their congenital heart disease during the first year of life and 1 patient had balloon dilation of aortic coarctation at 5 years of age. In all patients, moyamoya syndrome was diagnosed after surgical intervention for congenital heart disease-at 6 months of age in 1 patient, at 2 years of age in 3 patients, and at 6 years in 1 patient. Strokes were the most common presenting sign (3 patients) followed by seizures (2 patients). By the age of 33 months, 4 of 5 patients had undergone cerebral revascularization surgery to halt the clinical progression of moyamoya syndrome. CONCLUSIONS: Moyamoya syndrome should be considered in the differential diagnosis of seizures and stroke in patients with structural congenital heart disease. Prompt diagnosis and surgical management of the occlusive cerebral angiopathy should lead to improved neurological outcome in these patients. PMID- 9417152 TI - The effects of a high-protein, low-fat, ketogenic diet on adolescents with morbid obesity: body composition, blood chemistries, and sleep abnormalities. AB - OBJECTIVE: To evaluate the efficacy and metabolic impact of a high-protein, low carbohydrate, low-fat ketogenic diet (K diet) in the treatment of morbidly obese adolescents with initial weights of >200% of ideal body weight. METHODS: Six adolescents, aged 12 to 15 years, weighing an average of 147.8 kg (range, 120.6 198.6 kg) and having an average body mass index of 50.9 kg/m (39.8-63.0 kg/m), consumed the K diet for 8 weeks. Daily intake consisted of 650 to 725 calories, which was substantively in the form of protein (80-100 g). The diet was very low in carbohydrates (25 g) and fat (25 g). This was followed by 12 weeks of the K diet plus two carbohydrates (30 g) per meal (K+2 diet). MAIN OUTCOME MEASURES: Anthropometric data and blood and urine were collected at enrollment, during week 1, and at 4-week intervals throughout the course of the study. Resting energy expenditure was measured by indirect calorimetry. Body composition was estimated using dual-energy x-ray absorptiometry, bioelectrical impedance analysis, and urinary creatinine excretion at enrollment and on completion of each phase of the diet. Nocturnal polysomnography and multiple sleep latency testing were conducted at baseline and repeated after an average weight loss of 18.7 kg to determine sleep architecture, frequency and duration of apneas, and daytime sleepiness. RESULTS: Subjects lost 15.4 +/- 1.4 kg (mean +/- SEM) during the K diet and an additional 2.3 +/- 2.9 kg during the K+2 diet. Body mass index decreased 5.6 +/- 0.6 kg/m(2) during the K diet and an additional 1.1 +/- 1.1 kg/m(2) during the K+2 diet. Body composition studies indicated that weight was lost equally from all areas of the body and was predominantly fat. Dual-energy x-ray absorptiometry showed a decrease from 51.1% +/- 2.1% body fat to 44.2% +/- 2.9% during the K diet and then to 41.6% +/- 4.5% during the K+2 diet. Lean body mass was not significantly affected. Weight loss was accompanied by a reduction in resting energy expenditure of 5.2 +/- 1.8 kcal/kg of fat-free mass per day. Blood chemistries remained normal throughout the study and included a decrease in serum cholesterol from 162 +/- 12 to 121 +/- 8 mg/dL in the initial 4 weeks of the K diet. An increase in calcium excretion was accompanied by a decrease in total body bone mineral content. A paucity of rapid eye movement sleep and excessive slow-wave sleep were seen in all subjects at enrollment. Weight loss led to an increase in rapid eye movement sleep (P < .02) and a decrease in slow-wave sleep (P < .01) to near normal levels. CONCLUSIONS: The K diet can be used effectively for rapid weight loss in adolescents with morbid obesity. Loss in lean body mass is blunted, blood chemistries remain normal, and sleep abnormalities significantly decrease with weight loss. PMID- 9417153 TI - Implications of early sonographic evaluation of parapneumonic effusions in children with pneumonia. AB - OBJECTIVE: To devise a clinically relevant grading system for the sonographic evaluation of parapneumonic effusions, and to evaluate length of hospital stay as a function of treatment approach and sonographic grades. METHODS: Chest sonograms of 46 pediatric patients diagnosed with empyemas and admitted to two medical centers in the last 8 years were retrospectively evaluated using a grading system based on the degree of fibrinous organization within the parapneumonic effusions. Hospital charts were reviewed to determine the method of treatment and length of hospital stay. Patients were divided into two treatment groups: nonoperative (n = 26) (antibiotics alone, or combined with thoracentesis, or tube thoracostomy) and operative (n = 20) (open decortication, or video thoracoscopy and pleural debridement). Patients in the nonoperative group were further subdivided into two groups: those who received antibiotics alone (n = 11) and those who received antibiotics plus nonoperative drainage thoracentesis and/or tube thoracostomy (n = 15). Within each treatment group, patients were subdivided into two ultrasound grades: low (no evidence of organization) and high (evidence of organization such as fronds, septations, or loculations). Student's t test was performed to compare the lengths of hospital stay for each of the treatment groups and ultrasound grades. RESULTS: The length of hospitalization was no different for patients with low-grade ultrasounds in the nonoperative (9.8 days) and operative groups (8.0 days). In contrast, length of hospitalization was significantly shorter for patients with high-grade sonograms in the operative group (8.6 days), when compared with the nonoperative group (16.4 days). Length of hospitalization for patients in the nonoperative group with high-grade sonograms was significantly longer (16.4 days) than for those with low-grade ultrasounds (9.8 days). Furthermore, when the nonoperative patients were divided into an antibiotics alone group and a nonoperative drainage group, the patients with low-grade sonograms had no difference in the length of hospitalization (9.0 days vs. 10.4 days), whereas those patients with high-grade sonograms in the nonoperative drainage group had a significantly longer hospitalization (19.9 days) than the antibiotics alone (high-grade) group (11.4 days). CONCLUSIONS: Patients with a low-grade sonogram had similar length of hospitalization if treated with either nonoperative or operative measures. Patients with high-grade sonograms had significantly shorter length of hospitalization when treated with decortication. Our retrospective study suggests that patients with high-grade ultrasound studies treated nonoperatively do not benefit from pleural drainage procedures or chest tube placement. This study demonstrates the usefulness of early sonographic evaluation of parapneumonic effusions. A prospective study evaluating the usefulness of sonographic assessment of severity of disease in the treatment of children with parapneumonic effusions is warranted on the basis of our retrospective data. PMID- 9417154 TI - Lead levels in high-risk and low-risk young children in the Minneapolis-St Paul metropolitan area. AB - OBJECTIVES: To determine distribution of lead levels among children in a low-risk area; to validate a prescreening questionnaire; and to determine if universal lead screening is necessary in children in this area. DESIGN: Blood lead levels and questionnaires were obtained on eligible patients. Data were analyzed using stepwise regression analysis. SETTING: Community clinics and a health maintenance organization (HMO) in the Minneapolis-St Paul metropolitan area. PATIENTS: A total of 9603 children at well-child visits, age 6 months to 6 years at community clinics, and 6 months to 3 years at the HMO. OUTCOME MEASURES: Whole blood lead levels (WBLs) and questionnaires. RESULTS: The total sample rate of WBLs at >/=10 microg/dL was 12%, at >/=15 microg/dL was 31/2%, and at >/=20 microg/dL was 1.2%. At both 10 microg/dL and 15 microg/dL, the non-HMO group was at higher risk. For both groups, risk factors included living in the central cities, and living in housing built before 1950. For the non-HMO group a history of the child eating paint chips, or the child or a sibling having previous lead poisoning were also risk factors. CONCLUSIONS: Not all children need lead screening. Children living in the central cities, or with the risk factors of living in housing built before 1950 or a previous history of lead poisoning should be screened. PMID- 9417155 TI - The apgar score and its components in the preterm infant. AB - OBJECTIVE: The Apgar score is well-characterized in full-term infants but not in premature infants. The objective of this study was to assess the Apgar score in preterm infants with respect to the relationships between the 1- and 5-minute scores, the correlation of the Apgar score with pH and with other variables, and the relationship among the individual Apgar components. METHODOLOGY: We recorded Apgar scores at 1 and 5 minutes in a population-based cohort of preterm infants (n = 1105) with birth weight <2000 g, from three intensive care nurseries in central New Jersey. Linear correlation analysis was used to examine the relationship between 1- and 5-minute Apgar scores and between the individual components of the Apgar score. Multiple regression analysis was used to explore the relationship between various perinatal characteristics and the Apgar score, and between pH and Apgar score. Stepwise logistic regression analysis was used to assess the determinants of mortality. RESULTS: The 1-minute Apgar score median (25%, 75%) was 6(4,8) and correlated with the 5-minute score of 8(7,9) at r = .78. Slight but significant differences were seen between male (n = 557) and female (n = 508) infants in the 1-minute (6[4,8] and 7[4,8]) Apgar scores. One- and 5-minute scores of white infants (7[4,8] and 8[7,9]; n = 713) were significantly higher than those of black infants (5[3,7] and 8[6,9]; n = 280). Birth weight and gestational age were both linearly related to both Apgar scores. Low Apgar score (<3 at 1 minute and <6 at 5 minutes) was significantly associated with birth weight, gestational age and mode of delivery. Low arterial blood pH (<7.01) at birth was significantly related to low Apgar score. One hundred fifty nine infants died; these infants were significantly smaller (983 +/- 382 vs 1462 +/- 369 g), less mature (27 vs 31 weeks), had lower arterial blood pH (7.20 +/- 0.18 vs 7.31 +/- 0.11), had lower 1- (3[2,6] vs 7[4,8]) and 5-minute Apgar scores (6[4,8] vs 8[7,9]), and a greater incidence of low Apgar score (32% vs 6%) than did survivors. CONCLUSIONS: Among the components of the Apgar score, respiratory effort, muscle tone, and reflex activity correlated well with one another; heart rate correlated less well; and color the least. Our data confirms the limited use of the Apgar score in preterm infants and demonstrates the different responses of the Apgar score's components. PMID- 9417156 TI - Evaluation of a diagnostic approach to pediatric interstitial lung disease. AB - OBJECTIVE: To evaluate the value of a systematic approach to the diagnosis of pediatric interstitial lung disease (ILD). METHODS: In this descriptive, observational, prospective study, we evaluated 51 children presenting with ILD of unknown etiology during a 3-year period. Specific clinical information regarding history, physical examination, diagnostic evaluation, and final diagnosis was recorded on each patient. RESULTS: A specific diagnosis was established by history and physical examination alone in 1 patient, noninvasive tests alone in 8 others, and invasive tests, including lung biopsy, in another 26. Of the remaining patients, 8 had a suggestive diagnosis, and 8 had no specific diagnosis. CONCLUSIONS: A systematic approach to the diagnosis of pediatric ILD is useful, and not all patients need lung biopsy for diagnosis. PMID- 9417157 TI - Antistreptolysin O and anti-deoxyribonuclease B titers: normal values for children ages 2 to 12 in the United States. AB - BACKGROUND: Measurement of antibodies to the extracellular antigens produced by group A streptococci, antistreptolysin O (ASO) and anti-deoxyribonuclease B (anti DNase B), is often necessary to confirm a clinical diagnosis of a previous group A streptococcal infection, especially in patients suspected of having a nonsuppurative sequel to this infection. Age is among several factors that may influence antibody levels in children. Thus, in contrast to adults, what is considered a normal titer for one age group (infants) is not appropriate for another (older children). Age-related "normal" values for ASO and anti-DNase B are provided in the package inserts of commercially available kits; however, there are no recent comprehensive data to validate such values. OBJECTIVE: Using sera from 1131 children (from 23 states) ages 2 to 12 years, we determined age specific geometric mean titers (GMT) and upper limits of normal (ULN) of ASO and anti-DNase B. METHODS: ASO and anti-DNase B titers were measured by conventional laboratory methods. RESULTS: Children 7 years of age comprised the largest proportion (14%) of the study population. Approximately two-thirds of the sera were collected during winter and early spring months. For both ASO and anti-DNase B, both GMT values and ULN increased with age. The GMTs for ASO and anti-DNase B for the entire group of subjects were 89 and 112, respectively. The ULN for the entire group for ASO and anti-DNase B were 240 and 640, respectively. CONCLUSION: The age-specific values for GMT and ULN for this group of children from 23 states were slightly higher than previously reported. These values are likely representative of the pediatric population in the United States and should be of clinical value to physicians, epidemiologists, and clinical laboratory personnel. PMID- 9417158 TI - alpha1-Proteinase inhibitor therapy for the prevention of chronic lung disease of prematurity: a randomized, controlled trial. AB - BACKGROUND: An imbalance between increased neutrophil elastase and a decreased antiprotease shield has been suggested as a factor contributing to the development of chronic lung disease (CLD). We hypothesized that administration of alpha1-proteinase inhibitor (A1PI), also known as alpha1-antitrypsin, to premature neonates would prevent CLD. DESIGN: A randomized, placebo-controlled, prospective study of A1PI supplementation was performed. Neonates <24 hours of age with birth weights 600-1000 g on respiratory support, and 1001-1250 g with respiratory distress syndrome (RDS) were eligible. Intravenous A1PI (60 mg/kg) or placebo was infused on days 0, 4, 7, and 14. Primary outcome was CLD in survivors, defined as the need for supplemental oxygen on day 28. RESULTS: A total of 106 patients were recruited. There were no significant differences between groups in birth weight or incidence of RDS. The incidence of CLD in survivors was lower in the treated group, but the difference did not reach statistical significance (relative risk [RR], 0.79; confidence interval [CI], 0.60-1.02). This beneficial trend persisted at 36 weeks corrected gestational age (RR, 0.48; CI, 0.23-1.00). The incidence of pulmonary hemorrhage was lower in the treated group (RR, 0.22; CI, 0.05-0.98). Other complications were not significantly different between groups. CONCLUSIONS: In this, the first trial of a protease inhibitor for the prevention of CLD in premature infants, the infusions were well-tolerated. A1PI therapy may impede the development of CLD and appears to reduce the incidence of pulmonary hemorrhage in some neonates born prematurely. PMID- 9417160 TI - The epidemiology of overweight in children: relevance for clinical care. PMID- 9417161 TI - Exacerbation of autoimmune hepatitis: another hepatotoxic effect of pemoline therapy. PMID- 9417163 TI - Thalamic syndrome in children with measles infection and selective, reversible thalamic involvement. PMID- 9417162 TI - Fatal Kawasaki disease with coronary arteritis and no coronary aneurysms. PMID- 9417164 TI - Congestive heart failure in a neonate secondary to bilateral intralobar and extralobar pulmonary sequestrations. PMID- 9417165 TI - A school-based Chlamydia control program using DNA amplification technology. AB - OBJECTIVES: Chlamydia trachomatis is the most prevalent bacterial sexually transmitted disease (STD) in the United States, with the highest rates reported among adolescents. Chlamydia has severe consequences including pelvic inflammatory disease and infertility, and is believed to be a cofactor in human immunodeficiency virus transmission. Given that chlamydia is predominantly asymptomatic, most cases are identified through routine screening in health care settings. Over time, screening and treatment appear to be associated with a decrease in the prevalence of disease in areas with consistent chlamydia control programs. The new availability of sensitive and specific urine tests for chlamydia (polymerase chain reaction [PCR] and ligase chain reaction [LCR]) provides the opportunity to screen large numbers of at-risk youth in a noninvasive manner. We used PCR/LCR testing to investigate the feasibility of a school-based chlamydia control program and to determine the prevalence of chlamydia infection among junior and senior high school students. DESIGN: At three junior/senior high schools, all students, regardless of symptoms or sexual history, were given the opportunity to be tested for chlamydia using urine-based PCR or LCR testing. Only students with parental consent were eligible. Parents could not obtain test results, except if their children told them. During the five 3-week testing periods, throughout the day, classes were escorted to the testing area and each student was individually counseled regarding the opportunity to participate in the testing. SETTING: Three urban public schools in Louisiana. PARTICIPANTS: A total of 1933 students in grades 7 through 12, including 861 girls and 1072 boys. INTERVENTION: All students were informed about the test and taught about chlamydia during the homeroom period. Students were asked to provide a first-void urine specimen of not more than 30 mL. Specimens were refrigerated and delivered to the laboratory on the same day. Infected students were counseled and offered treatment with azithromycin, 1 g orally. They were also referred for or offered additional STD and human immunodeficiency virus testing. Infected students were asked to refer their sex partners to the city STD clinic for treatment. MAIN OUTCOME MEASURE: Prevalence of C trachomatis infection by grade and gender. RESULTS: Parental consent was obtained for 2849 (86.9%) of the 3278 matriculated students in grades 7 through 12. Fifty-one parents (1.6%) returned consent forms refusing permission for their child to participate in this screening and treatment program. The remaining 378 (11.5%) could not be reached by mail or telephone. Among all students with consent, 1933 (67.8% of those consented and 59.0% of those matriculated) were tested. Girls were less likely to be tested than boys (861/1363 [63. 2%] vs 1072/1465 [73.2%]). The overall prevalence of C trachomatis was 6.5%, with rates among girls more than twice that of boys (9.7% vs 4.0%). Generally, rates of infection increased with age. The prevalence rates among boys were for 7th grade, 2/208 (1%); 8th grade, 2/196 (2%); 9th grade, 10/236 (4.2%); 10th grade, 12/185 (6. 5%); 11th grade, 8/146 (5.5%); and 12th grade, 9/101 (8.9%). For boys 15 to 19 year old, the prevalence of chlamydia was 5.7%. Among girls, the prevalence rates were 7th grade, 0/105 (0%); 8th grade, 11/166 (6.6%); 9th grade, 23/218 (10.6%); 10th grade 23/146 (15.8%); 11th grade, 13/118 (11%); and 12th grade, 13/107 (12.1%). Among girls 15 to 19 years old, 12.7% were infected. Of 126 infected students, treatment was provided to 111 (88%). For this project, the laboratory cost of LCR testing was $17.76 per test. Without considering clinical staff time to collect the specimens, the average laboratory cost per infected student identified was $272. For students 15 to 19 years of age, of whom 104 (8.9%) of 1170 were infected, the laboratory cost was $200 per case identified. CONCLUSION: (ABSTRACT TRUNCATED) PMID- 9417167 TI - Hunger in children in the United States: potential behavioral and emotional correlates. AB - OBJECTIVE: Results from a recent series of surveys from 9 states and the District of Columbia by the Community Childhood Hunger Identification Project (CCHIP) provide an estimate that 4 million American children experience prolonged periodic food insufficiency and hunger each year, 8% of the children under the age of 12 in this country. The same studies show that an additional 10 million children are at risk for hunger. The current study examined the relationship between hunger as defined by the CCHIP measure (food insufficiency attributable to constrained resources) and variables reflecting the psychosocial functioning of low-income, school-aged children. METHODS: The study group included 328 parents and children from a CCHIP study of families with at least 1 child under the age of 12 years living in the city of Pittsburgh and the surrounding Allegheny County. A two-stage area probability sampling design with standard cluster techniques was used. All parents whose child was between the ages of 6 and 12 years at the time of interview were asked to complete a Pediatric Symptom Checklist, a brief parent-report questionnaire that assesses children's emotional and behavioral symptoms. Hunger status was defined by parent responses to the standard 8 food-insufficiency questions from the CCHIP survey that are used to classify households and children as "hungry," "at-risk for hunger," or "not hungry." RESULTS: In an area probability sample of low-income families, those defined as hungry on the CCHIP measure were significantly more likely to have clinical levels of psychosocial dysfunction on the Pediatric Symptom Checklist than children defined as at-risk for hunger or not hungry. Analysis of individual items and factor scores on the Pediatric Symptom Checklist showed that virtually all behavioral, emotional, and academic problems were more prevalent in hungry children, but that aggression and anxiety had the strongest degree of association with experiences of hunger. CONCLUSION: Children from families that report multiple experiences of food insufficiency and hunger are more likely to show behavioral, emotional, and academic problems on a standardized measure of psychosocial dysfunction than children from the same low-income communities whose families do not report experiences of hunger. Although causality cannot be determined from a cross-sectional design, the strength of these findings suggests the importance of greater awareness on the part of health care providers and public health officials of the role of food insufficiency and hunger in the lives of poor children. PMID- 9417166 TI - Eight-year outcome of universal screening and intrapartum antibiotics for maternal group B streptococcal carriers. AB - OBJECTIVES: To report the outcome of intervention to reduce early-onset group B streptococcal disease (EOGBSD) at a tertiary maternity hospital in Sydney and to review all cases of EOGBSD since intervention to improve outcomes further. METHODOLOGY: A prospective study was made of all cases of EOGBSD in the 16 months before and 8 years after an intervention that comprised universal screening and intrapartum ampicillin for all maternal carriers of group B streptococcus. Carriers were detected by screening all women at 28 weeks, or 24 weeks with known risk factors for preterm birth, by low vaginal swab, cultured onto blood agar and treated with intravenous ampicillin in labor, 1 g every 6 hours until delivery. Women with a routine midstream urine test positive for group B streptococcus, a previous neonate with EOGBSD, or preterm labor with an unknown carrier status were also treated. EOGBSD was detected by screening all neonates with maternal and/or neonatal risk factors for sepsis. RESULTS: The incidence of blood culture positive EOGBSD for all live births before intervention was 1.4 per 1000 compared with a rate after intervention of 0.2 per 1000 live births. The incidence, if there were clinical signs of infection and the urine tested positive for streptococcal antigen, decreased from 3.5 per 1000 before intervention to 0.6 per 1000 live births. There was a statistically significant reduction in neonatal morbidity outcomes after intervention, including requirement for admission and treatment in a neonatal unit and the need for ventilation. An audit indicated that by the 8th year, 90% of all pregnant women were screened by a low vaginal swab at 28 weeks and 10.5% were carriers. After intervention, of the 28 neonates with EOGBSD, 64% were associated with departure from the protocol. CONCLUSION: The intervention has coincided with a significant decrease in the incidence of blood culture-positive EOGBSD to 0.2 and urine streptococcal antigen-positive disease to 0. 6 per 1000 live births. The 84% reduction in EOGBSD has been obtained by treating 244 neonates in labor to prevent disease in one neonate. Additional reduction seems possible by improving the compliance by staff with the protocol. By contrast, before intervention and in maternity units throughout Australia with no intervention, the rates for EOGBSD remain largely unchanged at approximately 2 per 1000 live births. PMID- 9417168 TI - Newborn hearing screening: the great omission. AB - OBJECTIVE: The advent of technologic improvements in assessing the hearing of newborn infants has made possible the implementation of universal newborn hearing screening. Furthermore, selective screening based on high-risk criteria fails to detect half of all infants with congenital hearing loss. Although universal screening has been recommended by the National Institutes of Health and the Joint Committee on Infant Hearing Screening, data to support this recommendation have been incomplete, and the recommendation has been seen as without solid foundation by many in the pediatrics field. This study was designed to assess the feasibility, accuracy, and cost-effectiveness of a hospital-based hearing screening program for all newborns. METHODOLOGY: Between 1992 and 1996, hospitals in Colorado with 100 or more births per year were targeted to participate in universal hearing screening of newborns. To date, 26 of 52 targeted hospitals, ranging in size from 40 to 3500 births per year, have implemented universal screening. A total of 41 796 infants were screened between 1992 and 1996. Screening was performed using automated auditory brainstem response, otoacoustic emission testing, or conventional auditory brainstem response, with follow-up testing performed on those infants who failed initial screening. RESULTS: Of 41 796 infants screened at birth, 2709 failed initial screening, and of 1296 who have completed reevaluation, 94 have been identified with congenital sensorineural hearing loss (75 bilateral) and an additional 32 identified with conductive hearing loss (14 bilateral). The frequency of bilateral congenital hearing loss requiring amplification therefore is shown to be at least 1 in every 500 newborns. During the study period, an additional 17 children with significant hearing loss not identified until >/=18 months of age were reported voluntarily; all 17 had been born at hospitals not participating in newborn hearing screening. The false-positive rate for the screening program to date in Colorado is calculated to be 6%, but evolving technology has resulted in improvements to as low as 2%. Positive predictive value of an abnormal screen result is shown to be at least 5%, and as high as 19%, with improving technology. The sensitivity of newborn screening is demonstrated to be at or near 100%. Costs of screening are compared with other screened congenital diseases; although the true cost per child for newborn hearing screening is significantly higher than screening tests performed on blood, the much higher incidence of congenital hearing loss results in a comparable cost per case diagnosed when compared with hypothyroidism or phenylketonuria, for example. The feasibility of early intervention is demonstrated, with amplification by the use of hearing aids being the catalyst for effective treatment. Finally, the costs of screening and early intervention are compared with the monetary savings in avoiding delayed and therefore intensive therapy and intervention for children not diagnosed at birth. The true cost of screening for one newborn is shown to be between $18 and $33, with an average cost of $25 per infant. The cost per case of congenital hearing loss diagnosed is approximately $9600. A model for cost predictions and subsequent intervention savings is presented, and recovery of all screening costs is demonstrated after only 10 years of universal screening in Colorado. CONCLUSIONS: Universal newborn hearing screening is feasible, beneficial, and justified, as indicated by the frequency of the disease, the accuracy of screening tests, the ability to provide early intervention, the improved outcomes attributable to early amplification, and the recovery of all screening costs in the prevention of future intervention costs. Furthermore, the incidence of bilateral congenital hearing loss is alarming, and is, in fact, many times greater than the combined incidence of all newborn screening tests currently performed on blood samples. PMID- 9417169 TI - Does the supine sleeping position have any adverse effects on the child? II. Development in the first 18 months. ALSPAC Study Team. AB - OBJECTIVE: To assess whether the recommendations that infants sleep supine could have adverse consequences on their motor and mental development. DESIGN: A prospective study of infants, delivered before, during, and after the Back to Sleep Campaign in the United Kingdom, followed to 18 months of age. SUBJECTS: The children were participants of the Avon Longitudinal Study of Pregnancy and Childhood born to mothers resident in the three former Bristol-based health districts of Avon, with expected date of delivery from April 1, 1991 to December 31, 1992. Questionnaires were completed on sleeping position at 4 to 6 weeks of age and sets of standardized questions on development at 6 and 18 months. MAIN OUTCOME MEASURES: Social, communication, fine and gross motor, and total developmental scales based on the Denver Developmental Screening Test at 6 and 18 months. RESULTS: After adjustment for 27 factors using multiple regression, 3 of the 10 scales and subscales significantly distinguished between front and back sleeping position. At 6 months of age, infants put to sleep on their front had a mean score 0.38 SD (95% confidence interval [CI]: 0.28, 0.49) higher on the gross motor scale, 0.11 SD (95% CI: 0.00, 0.23) higher in the social skills scale, and a total development score 0.20 SD (95% CI: 0.10, 0.30) higher than those on their backs. These differences were no longer apparent at 18 months. CONCLUSIONS: There is some evidence that putting infants to sleep in the supine position results in a reduced developmental score at 6 months of age, but this disadvantage appears to be transient. Weighing this against the adverse health effects demonstrated with the prone sleeping position, these results should not change the message of the Back to Sleep Campaign. PMID- 9417170 TI - Fluoride supplement prescribing and dental referral patterns among academic pediatricians. AB - OBJECTIVE: To determine how well the current fluoride supplementation schedule was known by academic pediatricians and to examine the fluoride supplement prescribing and dental referral practices among primary care faculty pediatricians at four medical centers. SETTING: Four university medical centers in North Carolina. SUBJECTS: Primary care faculty pediatricians. METHOD: A questionnaire pretested for clarity was distributed to all identified full-time primary care pediatric faculty (42 members). RESULTS: A total of 40 completed questionnaires were returned. Thirty-seven (93%) primary care faculty pediatricians reported that they routinely addressed the need for fluoride supplements for their patients, but only 28 (70%) determined the fluoride content of the drinking water before prescribing supplements. Thirty-five (87.5%) began supplements at the correct age, but fewer knew the correct doses for children of various ages. Only 23 (58%) referred their patients for oral examination and preventive care before the age of 36 months, contrary to American Academy of Pediatrics recommendations. CONCLUSIONS: Pediatricians in an academic setting would be expected to be more knowledgeable of current recommendations than those in private practice. Failure to know and teach correct fluoride supplement recommendations and failure to recommend early professional dental involvement can result in less than optimum oral health. PMID- 9417171 TI - Is the incidence of appendicitis reduced in patients with sickle cell disease? AB - BACKGROUND: Patients with sickle cell disease (SCD) often present with abdominal pain, usually attributable to vasoocclusion. Experience at a single institution suggested that appendicitis was a rare cause of abdominal symptoms in this population. OBJECTIVE: We sought to determine whether the incidence of appendicitis was significantly lower in patients with SCD than in the population at large. METHODS: A 17-year retrospective chart review was performed at Rainbow Babies and Children's Hospital, Cleveland, OH, to determine the approximate incidence of acute appendicitis (AA) in patients with SCD. In addition, we performed a statistical analysis comparing the incidence of AA among SCD patients enrolled in the Cooperative Study of Sickle Cell Disease with that in the general population. RESULTS: Only two patients with SCD with pathologically confirmed AA were identified among approximately 200 patients followed at our institution during a 17-year period ( approximately 3500 patient-years), yielding an incidence rate of 5.7 cases per 10 000 patient-years. Among 3765 patients with SCD enrolled in the Cooperative Study of Sickle Cell Disease followed for a mean of 5.3 years (19 886 patient-years), a maximum of 9 cases of AA were identified, yielding an incidence rate of 4.5 cases per 10 000 patient-years. Based on data from the National Hospital Discharge Survey of 1978 to 1981, the incidence rate of AA in the general population (0 to 44 years of age) is approximately 16 per 10 000 patient-years. Paired t test analysis demonstrated a highly significant difference (P < .001) when comparing the incidence of AA among patients enrolled in the Cooperative Study of Sickle Cell Disease and the population at large. CONCLUSION: AA is an unusual event in patients with SCD. The likelihood of developing appendicitis in SCD patients is less than one third of that for the population at large. Conservative therapy is warranted in the large majority of patients with SCD who present with acute abdominal pain. Surgical exploration is best limited to patients with clear evidence of potential surgical pathology or progressive findings during a period of observation. The biologic basis of our findings remains unknown. PMID- 9417172 TI - Reasons for first teen pregnancies predict the rate of subsequent teen conceptions. AB - OBJECTIVE: To identify reasons for inconsistent contraceptive use that antedate conception and continue to predispose participants in adolescent-oriented maternity programs to unsafe sexual practices after delivery. We hypothesized that teens who attributed their failure to use contraceptives before their first conception exclusively to concerns about their side effects and/or their own lack of motivation to prevent conception would report less consistent contraceptive use and more repeat conceptions than would teens who attributed their previous failure to use contraceptives to their lack of capacity to do so. METHOD: We conducted a 2-year, prospective, longitudinal study of contraceptive use and repeat conceptions in a racially/ethnically diverse population of poor 13- to 18 year-olds. The 198 study participants were enrolled consecutively during their first pregnancies from an adolescent-oriented maternity program. RESULTS: The majority (84%) of the teens attributed their failure to use contraceptives before their first pregnancy partially to a lack of capacity to do so. As hypothesized, these teen mothers were significantly more likely to use hormonal contraceptives (85% vs 62%), (particularly Norplant, 47% vs 19%) and less likely to conceive again (13% vs 41%). Most teens attributed their inconsistent contraceptive use during the postpartum study period to three factors: side effects, plans to abstain from sexual intercourse, and their lack of motivation to postpone additional childbearing. CONCLUSIONS: The reasons teen mothers give for not using contraceptives consistently before their first pregnancies predict the occurrence of subsequent conceptions during adolescence. Those who attribute their previous failure to use contraceptives consistently to side-effect concerns and their own lack of motivation to postpone childbearing are least likely to use hormonal contraceptives after delivery and most likely to conceive again. Our findings suggest that future research should focus on the development of more effective interventions for preventing repeat conceptions among adolescent mothers who had the capacity to prevent their first pregnancies. PMID- 9417173 TI - Breastfeeding and later cognitive and academic outcomes. AB - OBJECTIVE: This study examines the associations between duration of breastfeeding and childhood cognitive ability and academic achievement over the period from 8 to 18 years using data collected during the course of an 18-year longitudinal study of a birth cohort of >1000 New Zealand children. METHOD: During the period from birth to age 1 year, information was collected on maternal breastfeeding practices. Over the period from 8 to 18 years, sample members were assessed on a range of measures of cognitive and academic outcomes including measures of child intelligence quotient; teacher ratings of school performance; standardized tests of reading comprehension, mathematics, and scholastic ability; pass rates in school leaving examinations; and leaving school without qualifications. RESULTS: Increasing duration of breastfeeding was associated with consistent and statistically significant increases in 1) intelligence quotient assessed at ages 8 and 9 years; 2) reading comprehension, mathematical ability, and scholastic ability assessed during the period from 10 to 13 years; 3) teacher ratings of reading and mathematics assessed at 8 and 12 years; and 4) higher levels of attainment in school leaving examinations. Children who were breastfed for >/=8 months had mean test scores that were between 0. 35 and 0.59 SD units higher than children who were bottle-fed. Mothers who elected to breastfeed tended to be older; better educated; from upper socioeconomic status families; were in a two parent family; did not smoke during pregnancy; and experienced above average income and living standards. Additionally, rates of breastfeeding increased with increasing birth weight, and first-born children were more likely to be breastfed. Regression adjustment for maternal and other factors associated with breastfeeding reduced the associations between breastfeeding and cognitive or educational outcomes. Nonetheless, in 10 of the 12 models, fitted duration of breastfeeding remained a significant predictor of later cognitive or educational outcomes. After adjustment for confounding factors, children who were breastfed for >/=8 months had mean test scores that were between 0.11 and 0.30 SD units higher than those not breastfed. CONCLUSIONS: It is concluded that breastfeeding is associated with small but detectable increases in child cognitive ability and educational achievement. These effects are 1) pervasive, being reflected in a range of measures including standardized tests, teacher ratings, and academic outcomes in high school; and 2) relatively long-lived, extending throughout childhood into young adulthood. PMID- 9417174 TI - Wernicke encephalopathy and beriberi during total parenteral nutrition attributable to multivitamin infusion shortage. AB - OBJECTIVE: Wernicke encephalopathy (WE) is an acute neurologic disorder characterized by a triad of ophthalmoplegia, ataxia, and mental confusion. WE is attributable to thiamine (vitamin B1) deficiency. Beriberi is the systemic counterpart of thiamine deficiency and often manifests in cardiovascular collapse. WE is usually associated with alcoholism and malnutrition. It has also been seen in people with gastrointestinal diseases with malabsorption. Patients who have received total parenteral nutrition (TPN) without proper replacement of thiamine have also developed WE. Since November 1996, there has been a shortage of multivitamin infusion (MVI). Many patients who were on chronic TPN with MVI ceased to receive the MVI and were converted to an oral form of the multivitamin. As a result, there have been several reports of children and adults on TPN who have developed WE as a result of thiamine deficiency. With this case report, we bring to attention the association of the MVI shortage and WE. Early diagnosis of WE is important, because if it is treated with thiamine in the acute stages, the neurologic and cardiovascular abnormalities can be reversed. CASE REPORT: We report a 20-year-old female patient with Crohn's disease who developed WE as a result of thiamine deficiency. She had Crohn's disease since age 9 years and was on chronic TPN. Two months before admission, MVI was discontinued in the TPN because of the shortage of its supply. An oral multivitamin tablet was substituted instead. She was admitted to the hospital for persistent vomiting. In the hospital, she continued to receive TPN without MVI, but continued taking an oral multivitamin preparation. Two weeks after admission, she developed signs of WE including diplopia, ophthalmoplegia, nystagmus, and memory disturbance. She also developed hypotension that was thought to be caused by beriberi. She was treated with 50 mg of intravenous thiamine. Within hours of the intravenous thiamine, her hypotension resolved. The day after the infusion, she no longer complained of diplopia, and her ophthalmoplegia had improved dramatically. Magnetic resonance imaging showed several areas of abnormally high signal on T2 weighted images in the brainstem, thalamus, and mamillary bodies. The topographic distribution of these changes was typical of WE. After 2 months, her mental status and neurologic status had recovered completely. CONCLUSION: WE and thiamine deficiency should be considered in all patients with malabsorption, malnutrition, and malignancies. WE from thiamine deficiency can occur as a result of cessation of MVI in the TPN infusion. Even if an oral multivitamin preparation is given instead of MVI, patients with malabsorption may not absorb thiamine adequately. Prompt diagnosis of WE is important because it is potentially fatal and readily treatable with thiamine supplementation. Early recognition of WE may be more difficult in children, because the classic triad of symptoms may not develop fully. Magnetic resonance imaging may be useful in these cases to confirm the diagnosis of WE. Because the shortage of MVI is expected to be a long-term, there are likely to be more cases of WE in the pediatric population of TPN dependent children. Because there is no shortage of intravenous thiamine, it should be administered with TPN even if MVI is not available. PMID- 9417175 TI - Cardiovascular malformations and complications in Turner syndrome. AB - BACKGROUND: Turner syndrome (gonadal dysgenesis with sex chromosome abnormalities) is recognized to be a disorder in which cardiovascular malformations are common. The prevalence and natural history of these findings, the risk for aortic dissection, and the occurrence of cardiovascular disease have all been the subject of debate, as have been the American Academy of Pediatrics recommendations for cardiac screening of patients with Turner syndrome. OBJECTIVE: To evaluate a large population of patients both cross-sectionally and longitudinally to determine the prevalence of cardiovascular malformations, the risk for dissection of the aorta, to determine whether there are phenotype:karyotype correlations that can allow for specific recommendations, and to devise an appropriate screening protocol. DESIGN AND METHODS: Data have been collected for patients with Turner syndrome. These individuals have been seen in an ongoing clinic established for the study of the natural history of Turner syndrome. Data from physical examinations, evaluations by cardiologists, echocardiography results, medical and surgical complications, medical records, and causes of death were analyzed. A total of 244 of 462 individuals in this population with karyotype-proven Turner syndrome could be evaluated because echocardiograms had been obtained. In addition, the medical literature was reviewed for occurrences of aortic dissection in patients with Turner syndrome. RESULTS: A total of 136 (56%) of 244 of these patients had cardiovascular abnormalities, 96 (71%) were structural, 40 (29%) were functional, including hypertension (HBP), mitral valve prolapse and conduction defects. Coarctation of the aorta and bicuspid aortic valve, alone or in combination, comprised >50% of the cardiac malformations. Bicuspid valve was often not detected by examination, but only by echocardiography. Aortic dissection occurred in three of the patients. In one, it was traumatic; in a second, it occurred at the site of coarctation repair. The third patient had long-standing HBP with malignant obesity. In the literature, there have been 42 case reports of aortic dissection in Turner syndrome. In all except 5, predisposing risk factors of coarctation, bicuspid aortic valve, and/or HBP were present. Of these 5, sufficient information regarding predisposing risk factors was provided for only 2. No phenotype:karyotype correlations could be drawn with any certainty. CONCLUSIONS: When the diagnosis of Turner syndrome is made, a screening echocardiogram should be obtained. Referral to a cardiologist first may be appropriate, but physical examination does not substitute for visualization. Individuals with and without evidence of structural cardiac malformations should be monitored for HBP on a lifelong basis. In the absence of structural cardiac malformations or HBP, the risk for aortic dissection appears small, and repeated echocardiography or magnetic resonance imaging to follow aortic root diameters does not appear to be warranted based on data currently available. Protocols for following patients with structural malformations need to be individualized, and wholesale recommendations have little merit. A longitudinal study using magnetic resonance imaging or cardiac echocardiography to establish normal parameters for aortic root diameters and to follow aortic root changes is needed. PMID- 9417177 TI - [Reconstruction of subtotal and total nasal defects]. PMID- 9417176 TI - Increasing prevalence of overweight among US low-income preschool children: the Centers for Disease Control and Prevention pediatric nutrition surveillance, 1983 to 1995. AB - OBJECTIVE: To determine whether the prevalence of overweight in preschool children has increased among the US low-income population. DESIGN: Analysis using weight-for-height percentiles of surveillance data adjusted for age, sex, and race or ethnicity. SETTING: Data from 18 states and the District of Columbia were examined. SUBJECTS: Low-income children <5 years of age who were included in the Centers for Disease Control and Prevention Pediatric Nutrition Surveillance System. RESULTS: The prevalence of overweight increased from 18.6% in 1983 to 21.6% in 1995 based on the 85th percentile cutoff point for weight-for-height, and from 8.5% to 10.2% for the same period based on the 95th percentile cutoff point. Analyses by single age, sex, and race or ethnic group (non-Hispanic white, non-Hispanic black, and Hispanic) all showed increases in the prevalence of overweight, although changes are greatest for older preschool children. CONCLUSION: Overweight is an increasing public health problem among preschool children in the US low-income population. Additional research is needed to explore the cause of the trend observed and to find effective strategies for overweight prevention beginning in the preschool years. PMID- 9417178 TI - [Intravenous gentamicin therapy in bilateral Meniere disease]. AB - BACKGROUND: The treatment of severe forms of bilateral Meniere's disease remains an especially challenging task. Similar problems also occur in debilitating Meniere's disease in the only hearing ear. The intramuscular titration therapy with streptomycin has been the means of choice since 1984 to minimize the risk of total hearing loss in cases of severe bilateral disease. METHOD: Since 1989 we have treated six out of 21 cases of bilateral Meniere's disease by intravenous application of 2 x 120 mg gentamicin in Ringer's solution for several days. Additionally we reported on two cases in 1988. Only minor amounts of gentamicin were applied to sedate the function of both vestibular organs while avoiding damage to the cochlea. RESULTS: In two cases hearing approved approximately about 10 dB, in two cases hearing remained stable, and in two cases hearing worsened about 10 dB. Five of six patients showed minor excitability in caloric tests on both sides, they did not complain of vertigo attacks one to five years after therapy. CONCLUSION: Given that only very small amounts of gentamicin are applied to sedate the function of the vestibular organ while causing almost no damage to the cochlea, this method seems to be an excellent means for treatment of bilateral Meniere's disease. Patients do not experience severe problems with equilibrium afterwards, and the treatment can be repeated as often as necessary. PMID- 9417179 TI - [Maculo-ocular reflex and visual perception during vertical acceleration]. AB - BACKGROUND: We investigated the effect of vertical acceleration upon the otolithic-ocular reflex of 22 healthy people. The study was performed to obtain standard values for subsequent investigations at patients. METHODS: People sitting on a chair were accelerated in the vertical axis with an amplitude of 4 cm and the frequencies of 0.5 Hz, 1 Hz, 1.5 Hz, 2 Hz, 2.5 Hz and 2.7 Hz. The movements of the ocular globe were initially recorded during vertical acceleration with the eyes closed. Then visual acuity was tested during linear acceleration with the eyes open. As parameters of evaluation we used coherence and alteration of the visual acuity. RESULTS: When the frequency was increased while the eyes were closed, coherence increased and the number of people with vertical eye movements increased. Amplitude was observed to increase and a phase shift occurred. A significant value of coherence (> 0.8) was observed at a frequency above 2.5 Hz. During the test of visual acuity, coherence also increased but did not reach quantity we observed initially. A significant loss of visual acuity occurred at a frequency above 2.5 Hz. A phase shift was also observed. The reason for the loss of visual acuity was the increment of amplitude and the phase shift, which had a negative influence on fixation. CONCLUSIONS: In summary, reactions with closed eyes are best tested at frequencies of 2.5 and 2.7 Hz. We recommended frequencies of 1.5 and 2 Hz for testing visual acuity. PMID- 9417180 TI - [Vestibular function: a prerequisite for normal language development?]. AB - BACKGROUND: First language acquisition depends on intermodal perception, especially auditive, tactile-kinesthetic, and partly visual in addition to sensomotoric integration. The influence of the vestibular function for a physiological language development is still unknown. PATIENT: A case history of a child with bilateral aplasia of all semicircular canals, normacusis in the right ear, and severe sensorineural hearing loss in the left (hearing aid supplied), without mental deficiency, allows us to draw initial conclusions. RESULTS: Logopedic diagnostics revealed only a mild articulation disorder (interdental sigmatism, inconstant gammacism) as a possible consequence of recurrent middle ear effusions since early childhood. Language development diagnostics did not demonstrate any significant norm deviation. CONCLUSIONS: In contrast to the statomotoric disorder, no evidence for a specific language impairment was found. Our report elucidates the importance of a selected diagnostic imaging (spiral CT and MIP MRI) in the phoniatric-ped-audiological field. PMID- 9417181 TI - [Orbital and intracranial complications of acute sinusitis in childhood--status of endoscopic paranasal sinus surgery based on case examples]. AB - BACKGROUND: Orbital or endocranial complications of sinusitis are rare clinical syndromes in infancy. METHODS: We retrospectively examined the complications that occurred and the role of functional endoscopic sinus surgery (FES). RESULTS: Most of the complications were caused by acute inflammation of the ethmoid or sphenoid cells with direct extension through dehiscent areas (flat bone) or neurovascular foramina. MRT had the highest sensitivity in detecting acute inflammatory foci. Early elimination of the cause of inflammation by FESS prevents spread of the complication. Advantages of FESS included better orientation and surgical access to the paranasal system of infants. The orbital complications were treated by surgical exploration of the communication pathways by a new endoscopic speculum. CONCLUSION: Early elimination of acute inflammatory foci by FESS prevents spread of infection in complications of paranasal sinusitis in infancy. PMID- 9417182 TI - [Long-term outcome after obliteration of the cat frontal sinus with ionomer bone substitute. II: Histomorphometric measurements]. AB - BACKGROUND: Stenosis or occlusion of the nasofrontal duct with subsequent recurrent frontal sinusitis or mucocele formation may require osteoplastic surgery in combination with frontal sinus obliteration. METHODS: To study its suitability as an implant material in sinuses, granular ionomeric cement was applied for frontal sinus obliteration in 15 cats. The frontal sinus specimens were processed for histological studies after 1, 3, 6, 12, and 24 months, respectively. To obtain quantitative data from the histological sections, areas of osteoid, connective tissue, and implanted ionomeric cement were measured with a computerized digitizing pad and compared to the total area of each frontal sinus. To our knowledge, histomorphometrical analysis of mesenchymal tissue reactions in close contact to an alloplastic material inside the frontal sinus cavity has not been described to date. RESULTS: Bone regeneration, starting from the sinus wall, was detected as early as one month after implantation. The quantitative data for osteoid indicated increasing osteoneogenesis and decreasing connective tissue growth inside the sinus cavity over the period of investigation. Two years after surgery, the osteoid represented 43.5%, the connective tissue 10.7%, and the ionomeric cement 45% of the whole cavity. The implanted cement did not show a significant degradation after two years. CONCLUSION: The results of the present study demonstrated the biocompatibility and biostability of the ionomer-base microimplant. PMID- 9417183 TI - [Tactile sensor for tissue differentiation in minimally invasive ENT surgery]. AB - BACKGROUND: In endoscopic surgery, stereoscopic vision and tactile information about tissue consistency are no longer available to the surgeon. METHODS: To compensate for these sensory deficits, various tissues can be characterized with an electromechanical sensor that records their resonance frequencies. In the future, the sensor will be integrated into surgical instruments, providing the surgeon with information about tactile properties of the tissue. We determined the impedance of tissues removed interoperatively (nasal polyps, lymph nodes, cartilage, bone) and different bony structures in a skull specimen. The examinations were carried out with an experimental setup and subsequently with a prototype of the tactile sensor. RESULTS: Resonance frequency increased with tissue hardness. Measurements with the experimental setup showed resonance frequencies for soft tissues between 15 and 30 Hz. We found that the bony septa of the ethmoid have a resonance frequency of 240-320 Hz and the thicker bony structures at the frontal skull base have a frequency of 780-930 Hz. Measurements of tumors in the upper aerodigestive tract showed that it is possible to differentiate between healthy mucosa, carcinomateous infiltrated mucosa, and carcinomateous undermined mucosa. In case of undermining tumor, the resonance frequency was one third higher than healthy mucosa. These results obtained with the experimental setup were reproduced with the tactile sensor prototype. CONCLUSIONS: The use of tactile information in endoscopic otolaryngological surgery may improve intraoperative tissue differentiation in the future. The safety of minimal invasive operations in head and neck surgery can be increased. PMID- 9417184 TI - [Therapy with nasal CPAP (continuous positive airway pressure) in patients with obstructive sleep apnea syndrome (OSAS). I: Long-term acceptance of nasal CPAP]. AB - BACKGROUND: Nocturnal ventilation with nCPAP has been established as the safest and most efficient nonsurgical treatment for OSAS. Long-term results, however, are determined by the patients' compliance with therapy. The aim of this study was the objective measurement of long-term acceptability of nCPAP therapy in all patients receiving this treatment in our sleep laboratory between January 1990 and March 1995. METHODS: We prospectively investigated 41 patients (36 male, 5 female) with moderate to severe OSAS who received nCPAP therapy. Mean time of follow-up was 20.6 months, ranging from 1.2 to 53.5 months. Therapy was indicated when OSAS was confirmed by cardiorespiratory polygraphy and either (1) the patient complained of daytime sleepiness or (2) the patient possessed an apnea hypopnea index greater than 30/h or when the mean oxygen desaturation was below 80% regardless of the presenting symptoms. The compliance with treatment was defined as a mean rate of use of over 5 hours per night calculated from the time counter on the nCPAP machine. RESULTS: 33 patients (88.5%) have continued using nCPAP until the present time but only 24 patients (59%) met our criteria for long term acceptance and this group was identified as responders. We found no significant differences in age, body mass index, apnea-hypopnea index, and nCPAP pressure between responders and non-responders. CONCLUSION: Although nCPAP is the safest treatment for OSAS, there is still a large group of patients with moderate to severe OSAS who are not efficiently treated with nCPAP because of the low long term acceptability of this therapy. With respect to this group of patients, surgical approaches have to be considered as an alternative therapy. PMID- 9417185 TI - [Detection of serum soluble fragments of cytokeratins 8 and 18 in patients with squamous epithelial carcinomas of the upper aerodigestive tract]. AB - BACKGROUND: During the development of squamous cell carcinomas of the head and neck (SCCHN), cytokeratin (CK) 18 and 19, which are typical for simple epithelia, show an elevated expression in tumor tissues. CK 8 was found to be overexpressed in cell lines of SCCHN. Epithelial tumors like normal epithelia express characteristic "pairs" of CK. The smallest of these pairs comprises CK 8 and 18. Therefore, the aim of this study was to serologically detect fragments of CK 8 and 18 (cyfra 8/18) in SCCHN patients. METHODS: Sera of 151 patients with SCCHN were tested for cyfra 8/18 (cyfra 8/18 ELISA medac, Hamburg) before treatment. Thirty sera of healthy volunteers and 43 sera of patients with benign diseases of the head and neck region served as control and reference group, respectively. Thirteen patients were followed-up clinically and serologically after surgery. The serum cyfra 8/18 concentrations of those 13 patients were compared to their clinical run. RESULTS: The cut-off value for cyfra 8/18 was determined with a 95% specificity of the reference group and was fixed at 1.1 ng/ml. The sensitivity was 7%. No correlation was found between cyfra 8/18 and clinical parameters like tumor stage, grading, and tumor location. Cyfra 8/18 concentrations could be correlated to the clinical run of 5 of 13 followed-up patients. CONCLUSIONS: Because of its low sensitivity, the serological detection of cyfra 8/18 must not be seen as a sufficient tumor marker for SCCHN. Its clinical applicability for regular post-surgical controls seems to be restricted and appears inferior to the clinical usefulness of cyfra 21-1. PMID- 9417186 TI - [Extramedullary plasmacytoma--manifestation in petrous bone]. AB - BACKGROUND: Extramedullary plasmocytoma of the temporal bone is a rare neoplastic disorder. Only a very few cases are described in the world literature. CASE: We report on a patient who presented with the clinical signs of a glomus jugular tumor. Postoperative histological evaluation revealed an extramedullary plasmocytoma of the temporal bone. CONCLUSION: If a glomus tumor is suspected, histological investigation appears necessary if surgery is not possible or is contraindicated. PMID- 9417187 TI - [The child with lip, maxillary, palatal cleft]. AB - Clefting of lip, alveolus and palate may occur in multiple variations. It causes aesthetic and functional detractions. Soft palate clefts may result in hearing-, speech- and swallowing-disorders. Therefore the otolaryngologist is a very important member in the interdisciplinary team directory. A cleft-palate child belongs to an interdisciplinary consulting hour in special hospitals, where different medical specialties are involved. Most important for a sufficient medical rehabilitation are maxillofacial surgery, otolaryngology, paediatrics, plastic surgery, speech therapy, psychology and human genetics. Also many other specialties may be involved. The cleft demands a complete follow up from the child's birth until it is grown up. Hearing disorders are caused by eustachian tube disfunction. There is a high prevalence of hearing loss and middle ear diseases in cleft palate patients. Hearing losses due to middle ear effusions in the very young child. Without therapy up to 50% of the cleft palate population will develop chronic middle ear diseases with and without cholesteatomas. Early and consequent therapy with myringotomy and insertion of a tympanostomy ventilation tube is necessary and helps to avoid chronic hearing problems. There is no general accepted system of speech disorders in cleft palate patients because of the difference in shaping of the cleft and rehabilitation development. Essential for speech rehabilitation are an intact velopharyngeal system and a keen sense of hearing. Both of it is disturbed in cleft palate children. Speech disorders are treated by speech therapists with prior consultation of the interdisciplinary team. The author presents a system of primary, secondary and tertiary speech disorders in cleft palate children. Primary speech disorders are caused by faulty velopharyngeal valving, offering in hypernasality, weak plosives, fricatives and affricates. Secondary speech disorders are substitute mechanisms for plosives, nasal and pharyngeal sounds. Tertiary speech disorders are hyper- and hypofunctional dysphonias following primary and secondary speech dysfunctions. PMID- 9417188 TI - [Interesting case no. 6. Chromophobe hypophyseal adenoma]. PMID- 9417189 TI - [On the 200th volume of the Zentrallblattes fur Hygiene und Umveltmedizin]. PMID- 9417190 TI - [Public health examination of water]. PMID- 9417191 TI - [Lighting the Munich residential theater with gas and with electric light]. PMID- 9417192 TI - [Traumatic conjunctivitis in miners]. PMID- 9417193 TI - [A comment on Dr. May's presentation: "Infectivity of milk from cattle with tuberculosis"]. PMID- 9417194 TI - [Infectivity of milk from tuberculosis infected cattle]. PMID- 9417195 TI - [Content of volatile fatty acids in various kinds of butter]. PMID- 9417196 TI - [Detection and toxicity of carbon monoxide and its presence in residences]. PMID- 9417197 TI - [Determination of vital fungi in the air]. PMID- 9417198 TI - Volumetric multiplexed transmission holography of the heart with echocardiographic data. AB - Three-dimensional imaging enhances delineation of cardiac anatomy and function. Currently, three-dimensional echocardiography involves complex rendering techniques to imply depth in a given image. Also, display of the final images on a video monitor neutralizes the volume information of the object. A hologram is a true, three-dimensional replica of the original object and does not entail complex data processing. Holography has not been used previously in cardiovascular imaging. In this experiment, 11 excised mammalian whole hearts and five isolated left ventricles were imaged in a water bath. Parallel, tomographic, echocardiographic images comprised the data source from which holograms were obtained by the technique of multiplexed holography. Holograms were mounted on a holographic film and viewed in a special viewing box. High-quality holograms were obtained from every data set. Excellent gray-scale discrimination allowed crisp visualization of cardiac structures and simulated diseases. Electronic sectioning and various projections optimized viewing of the three-dimensional anatomy in surgical orientations. Thus holograms can be produced from tomographic cardiac ultrasound data and could open a new avenue in three-dimensional echocardiography. PMID- 9417199 TI - Three-dimensional echocardiography: limitations of apical biplane imaging for measurement of left ventricular volume. AB - A new three-dimensional echocardiographic system creates a "line of intersection" display to allow precise and known positioning of echocardiographic images. Our purpose was to determine whether use of the line-of-intersection display will improve positioning of the apical four-chamber and apical two-chamber views and thereby improve the agreement between estimates of left ventricular volume by apical biplane echocardiography and cineventriculography. Unguided and line of intersection-guided apical biplane views were obtained in 31 patients immediately before cardiac catheterization and single-plane cineventriculography. In 15 patients the line-of-intersection display was used to measure the position of the image plane in studies of unguided and guided methods. Linear regression and limits of agreement analysis were used to assess the agreement between cineventriculographic volumes and echocardiographic volumes determined from each set of images. The Wilcoxon test was used to compare guided and unguided image positioning. The line-of-intersection display improved four-chamber and two chamber view positioning closer to the center of the ventricle and rotation closer to orthogonal positioning. Guided-image positioning was not able to correct displacement of the ultrasound beam anterior to the ventricular apex without deterioration of image quality in most patients. Despite improvements in image plane positioning, the agreement between echocardiographic and cineventriculographic volumes was unchanged. For end-diastole views, the unguided images had an r value = 0.84, standard error of the estimate of +/- 23.0 cc, and limits of agreement of +/- 62.4 cc. Corresponding values for the guided images at end diastole were r = 0.85, standard error of the estimate of +/- 22.9 cc, and limits of agreement of +/- 60.8 cc. At end systole the unguided results were r = 0.91, standard error of the estimate of 16.8 cc, and limits of agreement of +/- 52.2 cc. The line-of-intersection guiding of image plane positioning can improve apical image positioning but does not improve the agreement between apical biplane echocardiographic and cineventriculographic left ventricular volumes. The optimal apical imaging window is frequently occluded by the rib cage, resulting in a decrease in image quality. This reduction of image quality, combined with assumptions of left ventricular geometry, limit the accuracy of estimates of left ventricular volume from apical biplane echocardiography. PMID- 9417200 TI - Three-dimensional surface geometry correction is required for calculating flow by the proximal isovelocity surface area technique. AB - This study addressed the hypothesis that surface geometry must be taken into account in proximal convergence calculations of regurgitant flow rate. In vitro models allowed flow to converge within models designed to test derived angle correction equations. Flow was overestimated by the uncorrected equation for surfaces allowing flow to converge over less than a hemisphere and underestimated if flow converged over more than a hemisphere. The extent of deviation depended on the two-dimensional versus three-dimensional nature of the surface (angled flat surfaces versus conical surfaces). Correcting these estimates according to the derived equation produced good agreement for all geometries. PMID- 9417201 TI - Estimation of severity of stenosis with a Doppler guide wire in the experimental models. AB - Application of the continuity equation to Doppler catheter measurement of quantitative coronary flow velocity has been reported recently to be one of the accurate methods to evaluate mild to moderate coronary stenosis. This method, however, has not been validated in moderate to severe coronary stenosis. Furthermore, the ratio of prestenotic velocity/stenotic velocity may be influenced by side branches proximal to moderate to severe stenosis. Therefore we designed this study to evaluate the accuracy of the continuity equation method in the assessment of moderate to severe stenosis by an 0.018-inch (0.46 mm) Doppler guide wire (12 MHz) and the influence of a side branch on estimation of stenotic severity. Doppler spectra were recorded in the straight rigid tubes (4 mm diameter) with different severities of stenosis (50%, 62.5%, and 75% diameter stenosis) without a side branch with a Doppler guide wire. By the continuity equation, percent diameter stenosis was calculated from the proximal/stenotic and distal/stenotic velocity ratios in each model at different flow rates. In the model of 75% diameter stenosis with a side branch (1 mm diameter) proximal to the stenosis and the model of 75% diameter stenosis with a side branch (1 mm diameter) distal to the stenosis, percent diameter stenosis was calculated by the same method. Percent diameter stenosis derived from the proximal/stenotic velocity ratio in each model was in agreement with the true severity of stenosis in each model without a side branch (48% +/- 1%, 62% +/- 1%, and 75% +/- 1%, respectively). Percent diameter stenosis from the distal/stenotic velocity ratio was also in agreement with the true severity of stenosis in each model without a side branch (48% +/- 3%, 61% +/- 1%, and 75% +/- 1% respectively). In the model of 75% diameter stenosis with a side branch proximal to the stenotic site, however, percent diameter stenosis derived from the proximal/stenotic velocity ratio was underestimated compared with the real stenosis and significantly smaller than that derived from the distal/stenotic velocity ratio (65% +/- 1% versus 74% +/- 1%; p < 0.001). On the other hand, in the model of 75% diameter stenosis with a side branch distal to the stenotic site, the percent diameter stenosis derived from both the proximal/stenotic and distal/stenotic velocity ratios was in agreement with that derived from the distal/stenotic velocity ratio (75% +/- 2% and 77% +/- 1%). In the experimental models the estimation of stenotic severity by a Doppler guide wire is accurate even in moderate to severe stenosis unless a side branch exists just proximal to the stenosis. However, the distal/stenotic velocity ratio is applicable even if a side branch exists. This suggests that severity of stenosis derived from the distal/stenotic velocity ratio may be more useful than that from the proximal stenotic velocity ratio in human coronary stenosis with side branches. PMID- 9417202 TI - Echocardiographic and cardiac Doppler assessment of mice. AB - Recent studies have suggested that intermediate-frequency M-Mode transthoracic echocardiographic imaging is a promising method for evaluating the left ventricle in transgenic mice. However, there is a paucity of data regarding two-dimensional (2-D) echocardiography and cardiac Doppler echocardiography in this model. Therefore we studied 15 mice (body weights 38 to 65 gm) with an ultrasound system equipped with a 9 MHz transducer. M-mode, 2-D, pulsed, and color-flow Doppler studies were performed. Mean +/- SD for septal, posterior wall, and left ventricular cavity dimensions at end diastole were the following: M-mode: 1.1 +/- 0.2, 1.0 +/- 0.2, and 3.7 +/- 0.7 mm; 2-D: 1.0 +/- 0.2, 1.1 +/- 0.3, and 3.0 +/- 0.6mm. Left ventricular fractional shortening was assessed from the M-mode echocardiogram: mean 53.7% +/- 10.7% (range 42% to 77%). 2-D assessment of left ventricular mass correlated better with left ventricular mass identified at necropsy than left ventricular mass identified by M-mode echocardiography (r = 0.70; p = 0.007 versus r = 0.07; p not significant). 2-D visualization of left ventricle, proximal aorta, and aortic and mitral valves was excellent and was obtained mainly from a "parasternal" window. Apical views were more difficult to obtain. Mean +/- SD for aortic peak and mean velocities and velocity-time integral were 0.53 +/- 0.13, 0.32 +/- 0.08, and 0.025 +/- 0.008 m/sec. Estimated stroke volume was 0.0506 +/- 0.018 ml/beat. Cardiac output was 12.64 +/- 7.87 ml/min. Mean +/- SD for mitral peak E, peak A, and E/A ratio were 0.45 +/- 0.09 m/sec, 0.19 +/- 0.06 m/sec, and 2.4 +/- 0.66 m/sec, respectively. In all mice the E/A ratio was greater than 1 (range 1.76 to 3.6). Color-flowing imaging clearly displayed normal mitral inflow and left ventricular outflow. In one mouse, aortic regurgitation was recorded by pulsed Doppler echocardiography. Echocardiographic, pulsed, and color-flow Doppler assessment of mice is feasible. In this study left ventricular mass was assessed better by 2-D measurement of left ventricular dimensions. Assessment of left ventricular performance is feasible. Color Doppler guided evaluation of aortic flow and aortic root measurement permits assessment of stroke volume and cardiac output. PMID- 9417203 TI - Comparison of catheterization and Doppler-derived pressure gradients in a canine model of subaortic stenosis. AB - The relationship between Doppler-estimated and catheterization-measured pressure gradients was examined by repeated-measures linear regression analysis and difference plots in 15 dogs with naturally occurring subvalvular aortic stenosis. Thirty left ventricular outflow tract gradients were compared during sinus rhythm and 142 gradients during premature or postextrasystolic beats for the following pairs of data: (1) mean catheterization gradient versus mean Doppler gradient, (2) maximal instantaneous catheterization gradient versus maximal Doppler gradient, and (3) peak-to-peak catheterization gradient versus maximal Doppler gradient. The correlation between Doppler-derived and catheterization-derived pressure gradients was excellent (r = 0.99; p < 0.001) for the maximal instantaneous (sinus rhythm: standard error of the estimate [SEE] = 5.7 mm Hg; premature and postextrasystolic beats: SEE = 6.7 mm Hg) and mean gradients (sinus rhythm: SEE = 3.6 mm Hg; premature and postextrasystolic beats: SEE = 4.5 mm Hg). There was also a strong correlation between the peak-to-peak catheterization gradient and the maximal Doppler gradient (sinus rhythm: r = 0.99, p < 0.001, SEE = 5.3 mm Hg; premature and postextrasystolic beats: r = 0.97, p < 0.001, SEE = 7.2 mm Hg). Agreement between the two techniques was best for mean gradients and most disparate for the comparisons of maximal Doppler gradients and peak-to-peak catheterization gradients. PMID- 9417204 TI - Quantitative measurement of volume flow rate (cardiac output) by the multibeam Doppler method. AB - A new method has been developed for measuring the volume flow rate of blood flowing through large vessels or outflow tracts of the heart. In this article we describe the principle of a method that can reduce the dependence of the Doppler angle of flow measurement by setting the sample points along a line to which every ultrasound beam is perpendicular. To evaluate the accuracy of this method, flow phantom experiments were made for both steady and pulsatile flows. The volume flow rate measured by this method agrees well with that observed by an ultrasound flowmeter (r = 0.99) when the vessel diameter is large (25 mm). However, this method overestimates by 40% when the vessel diameter is small (8 mm). To make this method applicable to small vessels, an improvement in the lateral resolution of Doppler measurement is necessary. It has been concluded that this method can be used to measure the cardiac output or volume flow rates in large vessels. PMID- 9417205 TI - Dynamics of systolic pulmonary venous flow in mitral regurgitation: mathematical modeling of the pulmonary venous system and atrium. AB - The noninvasive assessment of mitral regurgitation has been an elusive clinical goal. Recent studies have highlighted the value of pulmonary venous (PV) flow reversal in indicating the presence of severe regurgitation. The purpose of this study was to explore the basic determinants of PV inflow in the presence and absence of regurgitation. In particular, the hypothesis that systolic PV flow depends on the interaction of regurgitant volume with atrial and PV properties (compliance, initial volume, total area of the pulmonary veins at the atrial junction, and the inertia of PV inflow) was tested and further, that the combination of these variables, rather than regurgitant volume alone, determines PV inflow. A mathematical model of the atrium and pulmonary veins was developed. Atrial and PV pressure were modeled as the product of chamber elastance and volume, where atrial elastance varied in time to simulate atrial relaxation and descent of the mitral anulus. A simplification of the modified unsteady Bernoulli equation was used to compute the PV velocities that resulted from the developed pressure gradient. The modeling was performed over a range of initial atrial elastances (0.77 to 0.2 mm Hg/cc), initial atrial volumes (20 to 75 cc), total PV areas (3.12 to 5.12 cm2), and PV inflow inertances (8 to 18 gm/cm2), with and without the addition of two regurgitant jets (regurgitant volume of 20 and 60 cc). The model realistically simulated the systolic PV waveform in magnitude and morphology. As the volume of regurgitation increased, PV peak flow velocity decreased, and eventually late systolic flow reversal occurred. However, the peak flow velocity, the time to peak flow, and the presence and magnitude of flow reversal were influenced by atrial compliance, volume, total atrial inlet area, and PV inflow inertia. This study found that PV flow blunting and reversal increased as atrial compliance, volume, and PV inertia decreased and as atrial inlet area increased. Atrial and PV properties (compliance, volume, total PV atrial inlet area, and PV inflow inertia), acting in combination, mediate the physiologic impact of the regurgitant lesion in terms of the resulting rise in atrial pressure as reflected by the pattern of systolic PV influx. For example, PV flow reversal is more likely in acute compared with chronic regurgitation because the atrium is less compliant and has a smaller initial volume. Therefore, the clinical assessment of mitral regurgitation using changes in systolic PV flow must be viewed in the context of atrial and PV properties. PMID- 9417206 TI - Pulmonary venous velocity patterns in mitral stenosis. AB - Pulmonary venous (PV) velocities obtained by transthoracic echocardiography are used to assess diastolic function. The systolic/diastolic (S/D) PV velocity ratio is increased with impaired early diastolic filling and decreased with elevated mean left atrial pressure (LAP). Mitral stenosis (MS) is characterized by impaired filling and elevated LAP. We hypothesized that the S/D ratio would be increased in MS except in severe MS with high LAP, in which the S/D ratio would be decreased. Patients with isolated MS who underwent transthoracic echocardiography were studied. The PV S/D ratio was compared in mild (n = 18), moderate (n = 16), and severe (n = 7) MS. There was no characteristic PV pattern, with each category showing a wide range of S/D ratios (mild, S/D ratio = 1.42 +/- 0.56; moderate, S/D ratio = 1.19 +/- 0.59, and severe, S/D ratio = 1.33 +/- 0.53) (mean +/- SD). Furthermore, there was no relationship between the S/D ratio and any index of severity of MS. There is no characteristic PV velocity pattern in MS. PMID- 9417207 TI - Measurement of aortic blood flow by Doppler echocardiography: temporal, technician, and reader variability in normal subjects and the application of generalizability theory in clinical research. AB - Although Doppler echocardiographic measurements of aortic flow have been found to correlate with stroke volume, the reliability of this technique is unknown. The purpose of this study was to measure the reliability of Doppler estimates of cardiac output by identifying and estimating the magnitude of different sources of error. We measured the reliability of Doppler estimates of cardiac output by identifying the magnitude of sources of error in 11 subjects with studies performed by two technicians and read by two readers. Analysis with generalizability theory demonstrated that the largest portion of the total variance was from differences among patients, with a smaller contribution due to day-to-day variability. Variability due to technician was low for continuous wave Doppler (2.0%), but high for pulsed wave (23.2%). Thus continuous wave, but not pulsed wave Doppler measurements, can be used to detect serial changes in cardiac output due to an intervention. PMID- 9417208 TI - Progressive intraventricular drop of early diastolic flow velocity reflects impaired active ventricular diastolic function in hypertensive heart disease: comparative study between early diastolic and atrial contraction phases. AB - Left ventricular (LV) diastolic function in the early diastolic phase includes both active and passive processes, but in the atrial contraction phase it includes only passive processes. To elucidate the relation between the intraventricular dispersion of the flow velocity in diastole and LV diastolic process, 31 normal volunteers and 12 patients with hypertensive heart disease were studied. In these subjects the flow velocity pattern at the mitral tip was recorded simultaneously with regional flow velocity patterns 1, 2, or 3 cm from the mitral tip toward the apex, respectively, with multigate pulsed Doppler echocardiography from the apical long-axis view with the guidance of Doppler color-flow imaging. Although the ratio of regional peak flow velocity/mitral peak flow velocity in the atrial contraction phase decreased from the mitral tip to the apex to the same degree in the normal volunteers and patients with hypertensive heart disease, there was a significant difference in the intraventricular dispersion of the early diastolic flow velocity between the two groups. These results suggest that the progressive intraventricular drop of the flow velocity in the early diastolic phase in patients with hypertensive heart disease may reflect the impairment of active rather than passive LV diastolic function. PMID- 9417209 TI - Doppler tissue imaging: myocardial wall motion velocities in normal subjects. AB - With a scanner modified for Doppler tissue imaging, mean myocardial velocities (MMV) across the myocardium were measured. The aim of this study was to determine the normal range of the maximum MMV in six standardized phases of the cardiac cycle. The MMV was defined as the average value of the myocardial velocity measured along each M-mode scan line throughout the thickness of the myocardium. The maximum MMV was defined as the maximum value of the MMV during the particular cardiac phase. Simultaneous gray-scale and Doppler tissue imaging M-mode images were taken of the interventricular septum and the left ventricular posterior wall from the parasternal long-axis and short-axis views in 15 normal volunteers (aged 21 to 47 years; mean 32 +/- 6 years). Each cardiac cycle was divided into six phases: atrial contraction, isovolumetric contraction, ventricular ejection, isovolumetric relaxation, rapid ventricular filling, and diastasis. Isovolumetric contraction, isovolumetric relaxation, and diastasis were subdivided into two parts a and b because of changes in the direction of the myocardial movement. For each volunteer, the mean and standard deviation of the maximum MMV were measured for each cardiac phase averaged from 12 cardiac cycles from both long-axis and short-axis views. Finally, the mean and standard deviation were taken for each cardiac phase from 180 cardiac cycles from 15 volunteers. We have found that specific cardiac phases show significant differences in the maximum MMV between the adjoining cardiac phases and significant differences also occur between the maximum MMV measured in the interventricular septum and the left ventricular posterior wall during the same cardiac phases. These normal values provide a standard against which future Doppler tissue imaging M-mode studies of abnormal left ventricular function might be compared. PMID- 9417210 TI - Pitfalls in creation of left atrial pressure-area relationships with automated border detection. AB - Creation of pressure-area relationships (loops) with automated border detection (ABD) involves correction for the variable inherent delay in the ABD signal relative to the pressure recording. This article summarizes (1) the results of in vitro experiments performed to define the range of, and factors that might influence, the ABD delay; (2) the difficulties encountered in evaluating a thin walled structure like the left atrium in the dog model; and (3) the solutions to some of the difficulties found. The in vitro experiments showed that the ABD delay relative to high-fidelity pressure recordings ranges from 20 to 34 msec and 35 to 57 msec at echocardiographic frame rates of 60/sec and 33/sec, respectively. The delay was not influenced significantly by the type of transducer used, distance from the target area, or size of the target area. The delay in the ABD signal, relative to the echocardiographic image, ranges from nil to less than one frame duration, whereas it is delayed one to two frame durations relative to the electrocardiogram processed by the imaging system. In the dog model, inclusion of even small areas outside the left atrium rendered curves with apparent physiologic contour but inappropriately long delays of 90 to 130 msec. To exclude areas outside the left atrial cavity, time-gain compensation and lateral gain compensation were used much more extensively than during left ventricular ABD recording. By changing the type of sonomicrometers used in our experiments, we were able to record simultaneously ABD and ultrasonic crystal data. However, both spontaneous contrast originating from a right-sided heart bypass pump and electronic noise from the eletrocautery severely interferred with ABD recording. PMID- 9417211 TI - Improved quantification of left ventricular function by applying signal averaging to echocardiographic acoustic quantification. AB - The acoustic quantification technique for on-line detection of endocardial boundaries currently provides continuous left ventricular area or volume signals and beat-to-beat ejection fraction. However, the distortion of individual waveforms by noise results in a wide beat-to-beat variability in these parameters. We developed an automated algorithm for the evaluation of left ventricular function by averaging acoustic quantification signals. End-diastolic and end-systolic area, stroke area, and fractional area change are measured directly from the average waveform. Peak ejection and peak filling rates and time to peak filling rate are obtained from its time derivative. Area signals obtained from eight normal subjects were used to evaluate the performance of this algorithm. Parameters of left ventricular function obtained with the automated algorithm were highly consistent and in excellent agreement with those obtained by repeated manual operator-dependent selections. This algorithm provides a fast and easy method for noise reduction in acoustic quantification signals, which significantly improves the noninvasive assessment of left ventricular function. PMID- 9417212 TI - Left ventricular isovolumic relaxation flow and left ventricular systolic performance. AB - We investigated isovolumic relaxation flow in patients with coronary artery disease (CAD) and evaluated the relationship between its velocity and left ventricular performance in 23 patients with atypical chest pain, 30 patients with CAD without prior myocardial infarction (MI), and 57 patients with prior MI, in whom cardiac catheterization was performed. The isovolumic relaxation flow velocity was measured at the basal portion of the left ventricle with pulsed Doppler echocardiography. The isovolumic relaxation flow ( > 15 cm/sec) was detected in 98 of 110 patients. The isovolumic relaxation flow velocity was significantly lower in patients with prior MI than in patients with atypical chest pain (p < 0.001) and in those with CAD without prior MI (P < 0.05). It was significantly lower in patients with CAD without prior MI than in those with atypical chest pain (p < 0.05). The isovolumic relaxation flow velocity showed a significant positive correlation with left ventricular ejection fraction. It also showed a significant negative correlation with left ventricular end-systolic volume index. These findings suggest that the isovolumic relaxation flow velocity is decreased in patients with CAD and is influenced by left ventricular systolic performance. Isovolumic relaxation flow may be a clinical manifestation of elastic recoil of the left ventricle. PMID- 9417213 TI - The comparative diagnostic value of dobutamine stress echocardiography and thallium stress tomography for detecting restenosis after coronary angioplasty. AB - The noninvasive detection of restenosis is clinically important for the subsequent management of patients in whom percutaneous transluminal coronary angioplasty (PTCA) has been performed. The aim of this study was to compare the diagnostic value of dobutamine stress echocardiography (DSE) and stress 201Tl single-photon emission computed tomography (SPECT) for detecting restenosis after PTCA. Fifty-three consecutive patients referred for the evaluation of possible restenosis or whom had been scheduled for follow-up study underwent DSE a mean of 5 months after angiographically successful PTCA. Dobutamine was infused incrementally under two-dimensional echocardiographic imaging. The left ventricle was divided into 16 segments and grouped into three coronary vascular territories. Rest, low-dose, and peak-dose images were digitized and displayed in a quad-screen format. Positive findings for restenosis were defined as new or worsened wall motion abnormality at a previously dilated vascular territory. All but one patient underwent SPECT. Positive findings for restenosis were defined as the presence of redistribution. All patients underwent quantitative coronary angiography after two tests. Restenosis was angiographically demonstrated in 23 (43%) of 53 patients and 25 (42) of 59 vessels. The sensitivity of DSE and SPECT for detecting restenosis was 78% and 74%, specificity was 93% and 93%, and accuracy was 87% and 85%, respectively. In a total of 59 vascular regions, DSE was 76% sensitive and 94% specific for detecting individual restenosis. It is concluded that DSE is comparable in diagnostic accuracy to SPECT for detecting restenosis in patients after PTCA. PMID- 9417214 TI - Predicting the severity of coronary lesions by the continuous recording method of exercise two-dimensional echocardiography. AB - We estimated the severity of coronary artery disease by the continuous-recording method of exercise two-dimensional echocardiography (Ex.2DE) in 56 patients with angiographically significant coronary artery stenosis ( > 50% diameter narrowing) who had undergone both Ex.2DE and coronary angiography. Patients were divided into two groups on the basis of findings of coronary angiography: group 1 had 50% to 89% stenosis (n = 24) and group 2 had 90% or greater stenosis (n = 32). The sensitivity and specificity of Ex.2DE for the detection of ischemic segments were 82% and 88%, respectively in the overall patient population. The sensitivity was 67% in group 1 and 94% in group 2. Hyperkinesis occurred at the beginning of exercise in 21 (88%) of 24 patients in group 1 and 15 (47%) of 32 patients in group 2 (p < 0.05). Our findings demonstrated that patients who did not show hyperkinesis at the beginning of exercise had more severe coronary artery disease. Careful observation of serial wall motion during exercise by the continuous-recording method may provide important information about myocardial ischemia. PMID- 9417215 TI - Intravenous perfluoropropane-exposed sonicated dextrose albumin produces myocardial ultrasound contrast that correlates with coronary blood flow. AB - If microbubble gas blood solubility and diffusivity are reduced, the persistence (and hence ultrasound reflectivity) of the microbubble in blood is prolonged. Recently we have sonicated a multifold dilution of human albumin with 5% dextrose while exposed to gases of low blood solubility and diffusivity and produced microbubbles that consistently opacify the myocardium after intravenous injection. The objective of this study was to test the hypothesis that a gas with very low diffusivity, perfluoropropane, when introduced into dextrose albumin during sonication, would produce visually evident myocardial ultrasound contrast after intravenous injection compared to sonicating with gases that have more rapid diffusivity. Second, we sought to determine whether the degree of contrast (peak myocardial videointensity) achieved with this agent would correlate with coronary blood flow. In eight open-chest dogs, intravenous injections of dextrose albumin sonicated with either room air, sulfur hexafluoride, or perfluoropropane (PESDA) were given under baseline conditions. PESDA injections were repeated when coronary flow was increased during low-dose dobutamine infusion. Left anterior descending coronary blood flow was monitored with transit-time flow probe. Background-subtracted anterior myocardial peak videointensity was measured after each injection. Visible myocardial opacification was seen in 100% of the 0.04 to 0.08 ml/kg intravenous injections of PESDA. No significant myocardial contrast was observed with the same doses of intravenous room air- or sulfur hexafluoride exposed sonicated dextrose albumin. There was a strong correlation between left anterior descending coronary artery flow (range 17 to 96 ml/min) and myocardial peak videointensity (r = 0.75; p < 0.0001) in all dogs. We conclude that intravenous injections of PESDA can safely produce consistent myocardial ultrasound contrast. the peak videointensity produced correlates with changes in coronary blood flow. Therefore this agent could be used to quantify coronary blood flow noninvasively. PMID- 9417216 TI - Intracoronary and aortic root myocardial contrast echocardiography: the effect of route, dose, and pharmacologic coronary vasodilation. AB - Myocardial contrast echocardiography is useful for the assessment of myocardial perfusion but has required direct intracoronary injections. Aortic root myocardial contrast echocardiography has the potential advantage of allowing simultaneous assessment of multiple perfusion beds, as well as evaluating competitive and collateral flows. This study assessed the safety and efficacy of intracoronary and aortic root injections of sonicated 5% human serum albumin (Albunex) with and without concomitant coronary vasodilation. Without vasodilation, 72% of intracoronary injections had optimal myocardial enhancement, compared with 21% of aortic root injections. For individual patients, significant dose-response relationships existed for both intracoronary and aortic root injections, although contrast intensity for a given dose varied between patients. Pharmacologic vasodilation resulted in significant increases in contrast intensity and in the incidence of optimal myocardial contrast after aortic root injections. Aortic root myocardial contrast echocardiography potentially allows the simultaneous assessment of multiple perfusion beds through a route somewhat less invasive than that of direct intracoronary injections. PMID- 9417217 TI - Assessment of postinfarction ventricular septal ruptures by transesophageal Doppler echocardiography. AB - Transthoracic Doppler echocardiography has been shown to be a sensitive modality for the diagnosis of acute septal ruptures after myocardial infarctions. Transesophageal echocardiography has been shown to improve diagnostic accuracy and image quality in many clinical settings. We performed transesophageal Doppler echocardiography in 10 patients with acute septal ruptures. Transesophageal echocardiography provided improved visualization of the rupture morphology (6 of 10 by transthoracic versus 10 of 10 by transesophageal imaging), better detection of multiple rupture sites (2 by transthoracic, 5 by transesophageal study) and better detail of the direction of shunt flow. On the basis of the transesophageal echocardiographic appearance, we propose that septal ruptures after acute myocardial infarctions be classified as simple or complex, consistent with pathologic criteria for left ventricular septal and free wall ruptures. Transesophageal echocardiography proved a useful and safe adjunct to transthoracic imaging, overcoming the technical limitations in these critically ill patients. PMID- 9417219 TI - The time required to perform pediatric transthoracic echocardiographic studies. AB - The purpose of this study was to determine the time required to perform and evaluate transthoracic pediatric echocardiographic procedures by two groups of pediatric cardiologists; one group was university based and the other group was based in a private practice. Methods consisted of measuring five periods including technician preparation time, technician procedure time, physician imaging time, physician review and reporting time, and physician time in interpretation of the results of the study to the parent or patient. The study evaluated data for 200 studies for the university-based group (all of which were complete studies) and 193 studies for the private practice group (84% of which were complete studies and the remainder were partial studies). Eleven pediatric cardiologists participated in the study. Although some variations in physician imaging and review time were encountered, data demonstrated that mean total physician time for a complete echocardiographic study was 31.9 minutes for both groups. Mean total technical time for complete studies was 42.4 minutes for the private group and 40.9 minutes for the university group. These are the first data evaluating the technical and physician work durations for pediatric echocardiography. PMID- 9417218 TI - Reproducibility of quantitative pediatric transesophageal echocardiography. AB - Transesophageal echocardiography (TEE) is commonly used to monitor cardiac function and to assess cavitary size. For the interpretation of quantitative echocardiographic data, the degree of their reproducibility should be considered. The variability of quantitative TEE was evaluated in this study. To assess intraobserver, beat-to-beat, interobserver, and repositioning variability, TEE examinations of 46 patients with congenital heart defects were analyzed. The mean beat-to-beat variability of 8.5% (range 4.2% to 12.3%) exceeded the mean intraobserver variability of 4.9% (1.9% to 8.1%). The mean interobserver difference between two observers was 3.4% (0.2% to 11.9%). Differences in image acquisition caused by repositioning of the transesophageal probe contributed the most (6.4% to 13.3%; mean 10.5%) to the variability of two-dimensional TEE. Changes seen on TEE studies should be interpreted as abnormal only when they exceed the total variability of this method. PMID- 9417220 TI - Multivariate analysis in the prediction of left atrial thrombi in patients with mitral stenosis. PMID- 9417221 TI - Left atrial thrombus formation immediately after cardioversion of atrial fibrillation despite adequate anticoagulant therapy. AB - We observed a patient who exhibited de novo left atrial thrombus formation after cardioversion, despite administration of adequate anticoagulant therapy. Preexisting atrial thrombus was excluded by transesophageal echocardiography. Preexisting severe left atrial mechanical dysfunction may be considered as a risk factor for de novo thrombus formation after cardioversion, as well as the poor outcome of cardioversion. PMID- 9417222 TI - Congenital sinus of valsalva aneurysm dissecting into the interventricular septum with left ventricular communication. AB - In this report we describe a case of a right coronary sinus of Valsalva aneurysm dissecting into the interventricular septum with spontaneous rupture into the left ventricle. Sufficient information was provided by echocardiography, cardiac catheterization, and aortography to confirm the diagnosis. Surgical findings were in complete accordance with cross-sectional and color flow Doppler imaging by transthoracic and transesophageal approaches. PMID- 9417223 TI - Coronary artery compression caused by a large pseudoaneurysm of the mitral-aortic intervalvular fibrosa. AB - An unusual late complication of bacterial endocarditis is described. Four years after diagnosis and treatment, a patient is seen with coronary artery compression caused by a large pseudoaneurysm of the mitral-aortic intervalvular fibrosa. The utility of transesophageal echocardiography in diagnosis and surgical management is emphasized. PMID- 9417224 TI - Left ventricular papillary fibroelastoma: two-dimensional echocardiographic detection and surgical resection. AB - We report a patient with a papillary fibroelastoma arising from the left ventricular posterior wall. The tumor was detected incidentally during echocardiography undertaken to evaluate aortic stenosis. Possible complication from tumor embolization was avoided by surgical resection during aortic valve replacement. PMID- 9417225 TI - Echocardiographic demonstration of intracardiac glue after endoscopic obturation of gastroesophageal varices. AB - Systemic embolism is an unusual complication of endoscopic obturation of gastroesophageal varices with glue. This report describes a case of cerebral embolism after this procedure. Intracardiac glue within the left atrium was demonstrated by echocardiography. Cardiac fluoroscopy demonstrated an abnormal vessel connecting periesophageal veins with the right upper pulmonary vein. Cardiac surgery was performed. Intracardiac glue was removed and the entering orifice of the abnormal vessel in the right upper pulmonary vein was sutured. To our knowledge, this is the first reported case of intracardiac glue after variceal obturation. Echocardiography is useful in the diagnosis of this rare complication. PMID- 9417226 TI - Absent right and persistent left superior vena cava without other congenital anomaly: a rare combination diagnosed by transesophageal echocardiography. AB - A 70-year old man with a history of anorexia, weight loss, and progressive shortness of breath was studied by transesophageal echocardiography. In addition to a mass occupying the right ventricular outflow tract, a rare congenital heart anomaly was discovered serendipitously: persistent left superior vena cava, absent right superior vena cava, and no other congenital abnormality. The echocardiographic findings were confirmed by computed tomographic scanning and later during heart surgery performed to resect the malignant tumor. PMID- 9417227 TI - Rupture of the left coronary cusp of the aortic valve caused by blunt chest trauma: early diagnosis by transesophageal echocardiography. AB - A case of a patient with a rupture of the left coronary cusp of the aortic valve caused by blunt chest trauma is presented. Early diagnosis by transesophageal echocardiography led to successful surgical repair. PMID- 9417228 TI - Another view of the use of multiple views. PMID- 9417229 TI - [Noninvasive and invasive monitoring of cerebral perfusion]. PMID- 9417230 TI - [Value of transcranial Doppler ultrasound for monitoring cerebral perfusion]. PMID- 9417231 TI - [Value of evoked potential monitoring in vascular surgery]. PMID- 9417233 TI - [Monitoring cerebral oxygenation: a methodological comparison]. PMID- 9417232 TI - [Noninvasive measurement of cerebral hemoglobin-oxygen saturation]. PMID- 9417234 TI - [Regional cerebral O2 saturation in hypothermia and total ischemia]. PMID- 9417235 TI - [Cerebral oxygenation (Hb) and oxidation status (cytochrome)]. PMID- 9417236 TI - [Measuring intracranial pressure]. PMID- 9417238 TI - [Evoked potentials--noninvasive indicators of intracranial pressure?]. PMID- 9417237 TI - [Intracranial pressure and anesthesia--EEG and p-EEG monitoring]. PMID- 9417239 TI - [Evoked potentials and intracranial pressure--experiences from the neurosurgical viewpoint]. PMID- 9417240 TI - [Degree of sedation and somatosensory evoked potentials in the recovery period after general anesthesia: a case report]. PMID- 9417241 TI - [Computer-assisted documentation of brain death with the Clipper program language]. PMID- 9417242 TI - [Hemodilution and brain protection?]. PMID- 9417243 TI - [Optimizing arterial CO2 and cerebral perfusion pressure in patients with craniocerebral trauma]. PMID- 9417244 TI - [Hypoxia protection of the blood-brain barrier: effects of barbiturates on hypoxic cultures of microvascular endothelial cells]. PMID- 9417245 TI - [Shunt placement in carotid operations?]. PMID- 9417246 TI - [Lysis therapy for improving cerebral reperfusion after heart arrest?]. PMID- 9417247 TI - [Biochemical brain protection]. PMID- 9417248 TI - [Brain protection in emergency situations: are anesthetics neuroprotective?]. PMID- 9417249 TI - [Current status of therapy in brain edema]. PMID- 9417250 TI - [Mild hypothermia and neuroprotection]. PMID- 9417251 TI - [Mild hypothermia (32 degrees C) improves cerebral oxygen balance during raised intracranial pressure--an animal experiment study]. PMID- 9417252 TI - [Anesthetic gas exposure at the work site--a solvable problem in the future]. PMID- 9417253 TI - [Pollution of the work environment by volatile anesthetics and nitrous oxide]. AB - Anaesthetic personnel is exposed to different workload conditions. The individual impact is influenced by external factors and human stress stability. Different symptoms reported to be present in anaesthetic personnel are comparable to symptoms of the sick building syndrome, defined by the WHO in the 90's. They are caused by work-induced distress and the exposure to chemical hazards. In anaesthesia, health defects by anaesthetic vapours and gases have been deplored for many years. After the Russian anaesthesiologist Vaisman published a report in 1967, controlled studies concerning cancerogenicity and teratogenicity of volatile anaesthetics under workspace conditions were carried out. In 1989, time weighted average exposure threshold limit values of 5 ppm were released in the Federal Republic of Germany for halothane. In 1993 thresholds for enflurane (20 ppm) and nitrous oxide (100 ppm) were released. TLV concentrations for the new anaesthetic agents desflurane and sevoflurane have not yet been defined by authorities. Factors influencing workplace concentrations of anaesthetic gases are the anaesthetic procedures, apparatus leakage, air conditioning, fresh gas flow and the function of the scavenging system. Although cancerogenicity, mutagenicity, teratogenicity and reduction of fertility are discussed as effects of chronic exposure to anaesthetic gases, several review articles doubted the results of studies, finding positive correlations of incidence of occupational disease and the exposure to the volatile and gaseous substances. Mainly coexisting factors like smoke-induced exposure to polybromated biphenyls, disturbance in circadian rhythm, stress and enclosure in narrow exposure systems, increasing teratogenicity and cancerogenicity in animal experiments, are considered to promote unreliability of the studies. All reviewers do not discuss the fact, that all of these co-factors are present in the reality of the anaesthetic workplace. Thus, the studies by Corbett, enthusiastically criticized by different reviewers, simulate the all-day reality of the anaesthetic workplace more precisely than controlled experiments conducted, for example, by Eger and co workers. The results of animal experiments and retrospective studies therefore do not justify realization of large controlled prospective studies but require the overall revision of the anaesthesiological workplace and the reduction of occupational waste gas exposure to the lowest possible levels below all chronic exposure threshold values. PMID- 9417254 TI - [4-chloro-m-cresol-induced contractures of skeletal muscle specimen from patients at risk for malignant hyperthermia]. AB - PURPOSE: 4-chloro-m-cresol (4-CmC), commonly used as preservative, has been shown to induce contractures in skeletal muscle specimens from individuals susceptible to malignant hyperthermia (MH). It has been suggested that a defect of the calcium release channel of the skeletal muscle sarcoplasmic reticulum (ryanodine receptor) in MH susceptible (MHS) patients could be responsible for this phenomenon. 4-CmC was found to be a potent activator of ryanodine receptor mediated Ca2+ release. The aim of this study was to determine the in vitro effects of 4-CmC on muscle specimens from MHS and normal (MHN) patients, and whether contracture testing with different concentrations of 4-CmC could result in a more precise discrimination between MHS and MHN. METHODS: In this prospective study muscle biopsies were obtained from 40 patients with clinical suspicion of MH. The patients were first classified by the in vitro contracture test (IVCT) according to the European MH protocol. After MH classification, surplus muscle specimens were subjected to the 4-CmC study. RESULTS: Cumulative administration of 4-CmC (25, 50, 75, 100, 150, and 200 mumol/l) produced contractures in a concentration-dependent manner. However, contractures developed significantly earlier and were greater in MHS (n = 17) than in MHN specimens (n = 23). After bolus administration of 50, 75, and 100 mumol/l 14-CmC MHS specimens developed distinct muscle contractures. In contrast, in MHN specimens only 100 mumol/l 4-CmC produced contractures. All contracture levels following bolus administration of 100 mumol/l 4-CmC were attained significantly earlier in MHS than in MHN. There was no overlapping in the range of times between both groups. CONCLUSION: In vitro contracture testing with 4-CmC seems to be a specific method to distinguish between MHS and MHN patients. However, the question whether 4-CmC is an MH-triggering agent is not completely solved. 4-CmC is a preservative within a large number of commercially available preparations (e.g. insulin, hormones, etc.). Regarding the results of contracture testing with 4-CmC it has been suggested that 4-CmC possibly represents a high-risk agent for MHS individuals. To reduce the risk of MH in susceptible patients due to administration of chlorocresols, we recommend avoiding preparations containing the preservative 4-CmC. PMID- 9417255 TI - [On-site laboratory monitoring on the intensive care unit. Blood gas, electrolyte, glucose, hemoglobin and lactate determination with the CIBA Corning 865 Analysis System]. AB - INTRODUCTION: For decision-making in the ICU, rapid and accurate analysis of vital laboratory parameters is essential. The industry provides devices which analyse these parameters on a decentralised setting and which are designed for use by non-laboratory personnel. We investigated whether accuracy and handling of a new analyser (Ciba-Corning 865, Chiron Diagnostics, Medfield, USA) are good enough for basing clinical decisions on the measured parameters. MATERIALS AND METHODS: The Ciba-Corning 865 allows measurement of blood gases, electrolytes, haemoglobin, glucose and lactate by use of photometric, ion-selective, enzymatic and electrochemical sensors in less than 18 microliters of whole blood. In a cardiac surgical intensive-care unit the accuracy of the device was tested by comparison to 61 measurements of quality control reagents, 48 tonometered blood samples and 536 parallel measurements in the clinical laboratory. Besides a 10 minute instruction, the participating personnel had no formal training with the device. RESULTS: The differences between measurements in quality control reagents and tonometered blood and the expected value were lower than 5%. The comparison with clinical laboratory measurements showed correlation coefficients from 0.94 (sodium) to 0.99 (glucose, lactate). The biases in Bland-Altman analyses were below 5%, the limits of agreement were found to be in a clinically acceptable range for all parameters. During the test period no technical problems occurred with the analyser and good acceptance by the personnel was found. CONCLUSIONS: The measured parameters were accurate enough to be used for therapeutic decisions in acute care medicine. Although it should not be a complete alternative to the clinical laboratory, because of rapid analyses, small sample volumes and easy handling the use of the Ciba-Corning 865 is advantageous for patients and users. PMID- 9417256 TI - [Anesthesia in general surgery and urology]. PMID- 9417257 TI - [False increased CK-MB value after cryoablation of the prostate without myocardial infarct]. AB - The authors report on a profound increase in creatine kinase isoenzyme MB (CK-MB) activity in three patients following uneventful cryoablation of the prostate under general anaesthesia: Just after the arrival at the recovery room CK-MB levels were 321 U/l, 245 U/l and 433 U/l, respectively. Other clinical investigations as well as additional laboratory tests ruled out myocardial infarction in all three patients. Electrophoresis of the CK-isoenzymes revealed an increase in CK-BB activity and an increase in atypical CK-BB as a cause of these findings. The presence of these isoenzymes leading to interferences with the antibody commonly used in CK-MB assays could explain the determination of a false positive CK-MB elevation in the three patients. Moreover, it is shown that this method of CK-MB activity determination may result in CK-MB levels higher than 100% of the whole CK activity. In addition, it is discussed that in patients suffering from prostatic carcinoma or other malignoma, "non-CK-M elevations" may occur. Therefore, the authors conclude that after cryoablation of the prostate additional tests like troponin T test and 12 channel ECG are required to rule out suspected myocardial infarction. PMID- 9417258 TI - [Therapeutic problems in tetanus--presented via a case report]. AB - This is the case presentation of a forty-year old female patient, who had incurred a tetanus infection as a result of intravenous drug abuse. Clostridium tetani could be detected repeatedly in abscesses caused by injections. The patient had to be put on continuous relaxation, sedation and artificial respiration for 42 days. Besides the usual intensive care regimen, a high-dose antitoxin therapy was initiated. The areas of abscesses had to be eradicated surgically several times. With the exception of a thrombus of the vena cava superior (without haemodynamic consequences) and a pneumonia, the further course was without any other serious complications. After seven months of hospitalisation the patient could be dismissed at "restitutio ad integrum". The known immunosuppressive effect of a high dosed tetanus antitoxin therapy could be confirmed by the patient's antitoxin titre course. Repeated active immunisation attempts to produce a sufficient endogenous antitoxin titre failed. The existing therapeutic uncertainties regarding the dosage of the tetanus antitoxin therapy, the titre control and the proper antibiotic treatment are described. PMID- 9417259 TI - [Echinococcus cysticus]. PMID- 9417260 TI - [Tumors of the craniocervical junction. I. Strategies of radiological examinations depending on the clinical syndrome]. AB - Tumors of craniocervical junction (ccjct) may cause a variety of non specific signs and symptoms. Before the advent of computed tomography (CT) diagnosis was sometimes possible only by surgical exploration, as the available radiologic methods--plain film radiography, angiography, and myelocisternography- essentially provided only indirect information about the neural structures. Because of its beam hardening artifacts, CT remained unsatisfactory as well. These diagnostic problems have practically disappeared with the availability of magnetic resonance imaging (MRI), which is now considered the method of choice if a space-occupying lesion is suspected at the ccjct. In some cases invasive angiography may still be necessary for surgical planning, and angiography may also be needed, whenever CT or MRI suggest the presence of an aneurysm. In the first part of this overview we present strategies for the radiological examination of the ccjct. PMID- 9417261 TI - [Tumors of the craniocervical junction. II. Morphological and radiological aspects with special reference to CT and MRT]. AB - Tumours of the craniocervical junction (ccjct) vary considerably in respect of survival times and cure rates. Correct localisation and tissue diagnosis are decisive for selecting the most suitable treatment. To do so one must be familiar not only with the clinical symptoms but also with the imaging features and the possible differential diagnoses of the individual tumour. In a second part of this overview we present the morphological and imaging features of the tumours occurring at the ccjct, and discuss in addition their epidemiological aspects. PMID- 9417262 TI - [Mammography and mammary ultrasonography: which examination sequence is preferable?]. AB - PURPOSE: To compare primary mammography diagnosis (ultrasound report available) with primary ultrasound diagnosis (mammography report available). METHODS: 89 preoperative patients with suspicious lesions were included. Mammography and ultrasound of all patients were evaluated by two independent experienced readers under clinical conditions. The reports of the complementary modality were available to both observers. Lesion evaluation was done on a per breast basis, in cases of multiple lesions in respect of the lesion with the greatest risk of malignancy. RESULTS: 39 benign and 59 malignant lesions were found. Primary mammography and primary ultrasound yielded 3 and 8 false positives and 10 and 13 false negatives. Concerning the palpable lesions (n = 59), primary mammography and primary ultrasound had no and 4 false positives and 7 and 8 false negatives, respectively, for the non palpable lesions, the figures were 3 and 4 false positive and 3 and 5 false negatives. CONCLUSIONS: Mammography remains the method of first choice in early detection of breast cancer, whereas breast ultrasound should be performed after and in knowledge of the mammogram, in consideration of the known indications (equivocal palpable lesion and mammographic opacity, dense breast). PMID- 9417263 TI - [Radiological changes after implantation of 2 different cementless hip prostheses]. AB - PURPOSE: Of the study was to evaluate radiological alterations of the femoral bone after implantation of two different prostheses. METHODS: 81 patients with 87 hips underwent total hip arthroplasty by using the Zweymuller-stem fixating in the medullary canal, and 175 patients with 182 ABG-stems with a proximally fixating, anatomical design were followed up clinically and radiologically. RESULTS: Adaptive bone remodelling in Zweymuller-stems showed in some patients thickening of the cortex of the femur and proximal bone-stock loss; reactive lines were seen in the proximal areas surrounding the prosthesis. Radiographs of patients with ABG-stems showed these lines in the distal, smooth part of the prosthesis and cancellous densification occurred in the area of load transfer distal to the trochanteric region. CONCLUSION: Typical x-ray findings due to various stem designs were identified and can be transferred to other cementless prostheses and should be considered if there is a suspicion of aseptic loosening. PMID- 9417264 TI - [CT angiography in arterial occlusive disease: comparison of 3 rendering techniques]. AB - PURPOSE: To evaluate different rendering techniques of CT data for the assessment of long vessel segments in peripheral vascular occlusive disease. MATERIAL AND METHODS: 40 CT angiograms (aortoiliac: n = 20, leg arteries: n = 20) were viewed using three different rendering techniques: 1, maximum intensity projection (MIP); 2, volume rendering (VR); 3, shaded surface display (SSD). CT angiograms were obtained in 6 or 8 projections. Axial cross-section images were analysed using an interactive cine mode. Intraarterial DSA was the standard in all cases. RESULTS: The sensitivities for the diagnosis of occlusive disease were 100% (cross-section images), 94% (MIP), 91% (VR) and 93% (SSD). The specificities were 100%, 99%, 99% and 99%, respectively. For the accurate grading of high-grade (> 75%) stenoses, the sensitivities were 85% (cross-section images), 62% (MIP), 44% (VR) and 35% (SSD). Specificity was 99% for all techniques. CONCLUSIONS: CTA is accurate in occlusive disease. Interactive viewing of cross-section images is the most accurate technique. MIP is superior to VR in the imaging of high-grade stenoses because contrast-to-noise ratio is high and thresholding is not necessary. PMID- 9417266 TI - [MRI of the regions of the inner ear and cerebellopontine angle using a 3D T2 weighted turbo spin-echo sequence. Comparison with conventional 2D T2-weighted turbo spin-echo sequences and T1-weighted spin-echo sequences]. AB - PURPOSE: To assess the value of a three-dimensional (3D) T2-weighted turbo spin echo sequence (3D T2-TSE) in comparison to conventional two-dimensional (2D) T2 weighted TSE and unenhanced and enhanced T1-weighted spin-echo sequences (SE) in imaging anatomic structures and pathologic changes of the inner ear and cerebellopontine angle. PATIENTS AND METHODS: The inner ear and cerebellopontine angle were investigated by MRI in three healthy volunteers and 18 patients performing a 2D T2-weighted turbo spin-echo sequence and a 3D T2-TSE in the axial plane. In the patient study, 2D T1-weighted SE sequences both before and after the i.v. injection of gadopentetate dimeglumine in both the axial and coronal plane were performed in addition. RESULTS: Only the 3D T2-TSE enabled an accurate imaging of the anatomic structures. In cases of pathology, the 3D T2-TSE provided additional information to the performed 2D sequences. The combination of the 3D T2-TSE with unenhanced and enhanced 2D T1-weighted SE enabled the most accurate diagnosis in cases of pathology. CONCLUSIONS: Accurate depiction of anatomic structures of the inner ear and cerebellopontine angle could be obtained by 3D T2 TSE only. The most accurate diagnosis in cases of pathology was provided by the combination of the 3D T2-TSE with unenhanced and enhanced 2D T1-weighted spin echo sequences. PMID- 9417265 TI - [Bolus tracking and NaCl bolus in biphasic spiral CT of the abdomen]. AB - PURPOSE: To optimise injection parameters in helical CT of the abdomen with individual bolus tracking and subsequent NaCl bolus injection. To investigate the effect of bolus tracking on image quality in the abdomen. METHODS: Patients were randomised into three examination protocols and underwent biphasic helical CT (Hi Speed Advantage, GE). The effect of NaCl bolus on duration of aortic enhancement was investigated. Contrast enhancement in parenchyma and vessels was examined. The influence of body mass index, injection flow and contrast material volume on enhancement was evaluated. RESULTS: Subsequent injection of NaCl provided significant extension of contrast enhancement in the aorta. Optimal image quality for pancreas and abdominal arteries was achieved in the arterial phase and for liver, spleen, kidneys and abdominal veins in the portal venous phase. Body mass index, injection flow and contrast material volume showed a significant influence on the time intervals resulting from bolus tracking. CONCLUSION: Individual bolus tracking with subsequent injection of 20 ml NaCl bolus optimises intravenous contrast application. PMID- 9417267 TI - [Contrast-enhanced MR cholangiography in percutaneous bile duct drainage]. AB - PURPOSE: To evaluate MR-cholangiography after instillation of contrast media via indwelling biliary tubes. METHODS: In 8 patients with stenoses of the central bile ducts, physiological saline solution and diluted contrast media (Gd-DTPA, 5 mmol/l) were consecutively administered via an indwelling biliary tube. MR cholangiograms were obtained before and after saline injection using a HASTE sequence and after administration of Gd-DTPA using a T1-weighted gradient echo sequence. RESULTS: Peripheral bile ducts were better visualised in the water sensitive approach than after Gd-DTPA enhancement. In patients with short-time drainage bile duct definition was poor due to periportal oedema. Visualisation of peripheral bile ducts could be improved by injection of saline solution in these patients. After administration of Gd-DTPA via the biliary tube contrast enhancement of the central bile ducts was achieved in all patients except for one patient on long-term drainage with an indwelling Yamakawa tube. CONCLUSION: T1 weighted visualisation of the central bile ducts can be achieved by means of injection of Gd-DTPA via indwelling biliary tubes. PMID- 9417268 TI - [MR enteroclysis for nuclear spin tomographic diagnosis of inflammatory bowel diseases with contrast enhancement]. AB - PURPOSE: To evaluate MRI for effectiveness in assessment of intra- and extramural changes in the small intestine. METHODS: 40 patients with known or suspected small bowel disease underwent MR imaging immediately after conventional enteroclysis with barium and a mixture of methyl cellulose and gadolinium-DTPA. RESULTS: In 6 of 24 patients with no pathological findings in conventional enteroclysis, intraabdominal pathology such as thickening of the intestinal wall and an abscess were identified. In the remaining patients, MRI showed good correlation with conventionally obtained data and provided important additional information regarding extraluminal involvement such as enlargement of mesenterial lymph nodes and fistulas as well as abscesses. CONCLUSIONS: MRI, carried out using this technique, provides important additional information regarding intra- and extraluminal changes with good image quality. PMID- 9417269 TI - [Doubtful mammographic findings: the value of negative MR mammography for tumor exclusion]. AB - PURPOSE: To determine whether the addition of MR mammography (MRM) is useful in excluding malignant lesions and how reliable negative MRM findings are. METHODS: Amongst 694 MRM's, those originally regarded as normal were retrospectively reappraised. 239 female patients were involved. In all these patients there were clinical, sonographic and/or mammographic findings which were not entirely normal but there was no urgent indication for histological clarification. In 46 patients there were, however, histological examinations since the patients themselves insisted on it. In the remaining patients there was clinical, sonographic, mammographic and/or MRM follow-up after 12 to 18 months. RESULTS: In 95.4% (200/239) a carcinoma could be excluded by means of MRM, in 7 patients a carcinoma in situ and in two patients an invasive carcinoma was demonstrated histologically which had not been demonstrated by MRM. Even in retrospect, no abnormality could be found. CONCLUSION: Because of the only moderate sensitivity of MRM in the recognition of carcinoma in situ, doubtful lesions which can be localised, should be biopsied by a stereotactic method. In cases where evaluation is difficult on clinical, sonographic and mammographic findings, MRM is of value in excluding tumours, particularly in patients with increased carcinoma risk. PMID- 9417270 TI - [MR tomography studies of myocardial function and perfusion after myocardial infarct]. AB - PURPOSE: With the advent of fast pulse sequences, MR imaging of myocardial function and perfusion in ischaemic heart disease has become possible. Prior studies examined either myocardial perfusion or systolic wall motion. We intended to establish an examination procedure to simultaneously investigate regional myocardial motility and perfusion in patients 7-14 days after myocardial infarction. METHODS: A Turbo-FLASH 2D sequence was optimised to maximise image contrast between normal and malperfused myocardium after Gd-injection using a calculation model basing on the Bloch equation. Calculated values for trigger delay TD, inversion time TI and flip angle alpha were confirmed in a Gd-phantom and healthy volunteers. Subsequently, myocardial motility was studied (cine FLASH 2D sequence) and in slice positions with reduced wall thickening first pass and post contrast studies after 2-10 minutes were performed using the optimised Turbo FLASH sequence. RESULTS: First pass SI-differences of normal compared to malperfused myocardium vary in relation to TD, TI and alpha in a relevant degree. Reduced myocardial motility was found with a sensitivity of 81% and specificity of 96%. Pathological perfusion patterns were detectable in all of these patients. CONCLUSIONS: A combined examination of motility and perfusion is possible by means of MRI and information about the status of postinfarct myocardium can be obtained. PMID- 9417271 TI - [Does the core biopsy of solid liver lesions permit an exact histological classification? Results of a prospective study under routine clinical conditions]. AB - PURPOSE: The aim of this prospective study was to answer the question to what extent percutaneous core-cut biopsy of solid lesions of the liver can permit (sub )classification under routine conditions. MATERIAL AND METHODS: Subject of this study were 80 percutaneous core-cut biopsies of solid liver lesions in 75 patients. The biopsies were done consecutively under routine conditions by 10 different radiologists with Tru-Cut-needles and a biopsy gun with sonographic (n = 73) or CT (n = 7) guidance. After receiving the histological analysis, the radiologists then prospectively divided the biopsies into "valid" und "unclear" using clinical and radiologic data of the respective patient. The results were verified by a second biopsy, follow-up or surgery. RESULTS: 80% of all histological analyses were rated as "valid", 20% as "unclear". The accuracy of the "valid" reports in respect of ranking was 95% and 90% for the accurate tumour classification. These values were reduced to 81.3% and 74%, respectively, with regard to the total number of cases. The negative predictive value was reduced from 78% to 51.7%. CONCLUSION: Core-cut biopsy of solid liver lesions guided by imaging yields good results even under routine conditions as long as all histological reports have been correlated with imaging and clinical informations. PMID- 9417272 TI - [Midterm follow-up after Cragg stent placement in iliac arteries]. AB - PURPOSE: Evaluation of midterm success and patency rates after placement of Cragg stents in iliac arteries. MATERIAL AND METHODS: During a period of 26 months 16 patients, with a total of 19 iliac lesions were treated percutaneously by placement of 20 stents. Indications of stent placement were iliac occlusion in 4 cases and high-grade iliac stenoses in 15 cases. Indication for stent placement in the stenotic lesions were insufficient results following balloon angioplasty in 11 cases and extensive dissection in 4 cases. RESULTS: The ankle-brachial index was improved from 0.53 +/- 0.28 to 0.85 +/- 0.26 immediately after the intervention and was 0.80 +/- 0.15 at 16 months follow-up. Cumulative patency rate was 71% after 12 months. CONCLUSION: The occlusion and restenosis rates are high. Disadvantages of Cragg stents are low flexibility and a large bore introducer system. The radiopacity of the Cragg stent is advantageous for fluoroscopic positioning. PMID- 9417274 TI - [Cystic degeneration of the adventitia of the popliteal artery: ultrasound angiography-aided puncture]. AB - A filiform stenosis of the popliteal artery was examined by sonography and the reason for the cystic lesion in the arterial wall was found to be an uncommon type of cystic degeneration of the adventitia. The narrow lumen could be demonstrated by sonography and the lesion was punctured under ultrasound control. A viscous secretion was removed and the patient's claudication was cured without recourse to surgery. PMID- 9417273 TI - [Cava filter placement under MRI control. Experimental in vitro and in vivo studies]. AB - PURPOSE: An instrument has been developed for the introduction of caval filters which can be used with MRI; it has been investigated in in vitro and in in vivo experiments. MATERIAL AND METHOD: The ferromagnetic components of a commercially available instrument for the femoral introduction of the MR-eye Tulip IVC filter were changed for similar, non-ferromagnetic parts and the lock and dilator marked with dysprosium oxide rings. The instrument was used in a flow phantom and in animal experiments (two domestic pigs) in order to insert filters under MRI control on a 1.5 T Philips Gyroscan with integrated mobile digital subtraction angiography. RESULTS: Both in vitro and in vivo, the introducer, catheter and caval filter could be identified by MRI and positioned under MRI control. The position of the filter as indicated by MRI corresponded with radiological and macroscopic findings in all cases (5 phantoms, 2 pigs). CONCLUSION: The early experimental results indicate that percutaneous introduction of caval filters with placement under MRI control is possible. PMID- 9417275 TI - [Demonstration by spiral CT of an aneurysm of the left sinus of Valsalva]. PMID- 9417276 TI - [MR tomographic and clinical course of intracochlear acoustic neurinoma]. PMID- 9417278 TI - [Leiomyosarcoma of the inferior vena cava]. PMID- 9417277 TI - [Placement of a coated stent in esophagotracheal fistula: initial experience with an "aortic stent"]. PMID- 9417279 TI - [BCG: substrains, the vaccine, vaccination]. PMID- 9417281 TI - [Hepatitis C viral infection. I. The premises in the discovery of the existence of the hepatitis C virus]. PMID- 9417280 TI - [Serological diagnosis in HIV infection]. PMID- 9417282 TI - [Hepatitis C viral infection. II. Viral morphology not yet determined]. PMID- 9417283 TI - [Education for health--smoking--testing the knowledge and behavior of some socio occupational groups--students, parents, professors--from Lucian Blaga High School in Sebes, Alba]. PMID- 9417284 TI - [Legionellosis in Spain--clinico-epidemiological data]. PMID- 9417285 TI - [The use of CAMP disks in demonstrating the pathogenicity traits of opportunistic Vibrionaceae]. PMID- 9417286 TI - [The demonstration of the production of "slime"--a marker of pathogenicity in coagulase-negative staphylococci]. AB - The authors tested for "slime" elaboration 48 S. epidermidis s.s. strains; 24 of these strains were isolated from bloodstream infections and 24 from resident cutaneous microbiota. The semiquantitative method of Christensen was used for testing "slime" elaboration. The predictive value of the positive test of "slime" elaboration was 92% in this study. PMID- 9417287 TI - [The sporicidal activity of chemical products tested in our laboratory on Bacillus subtilis spores]. PMID- 9417288 TI - [Glutaraldehyde used as a chemical agent in the disinfection and sterilization of thermolabile medical equipment]. PMID- 9417290 TI - [The T-lymphocyte]. PMID- 9417289 TI - [The pathogenesis of viral hepatitis C]. PMID- 9417291 TI - [The usefulness of demonstrating the presence of antimycobacterial antibodies. A comparative study of 2 different technics (ELISA and Myco-Dot)]. PMID- 9417292 TI - Not all self help groups discourage sick doctors from being members. PMID- 9417293 TI - [The Urologic Cooperative Group of the EORTC. Structure, scope, research, results]. AB - The European Organization for Research and Treatment of Cancer (EORTC) was founded in 1962. The first urological group was French-speaking and concentrated particularly on testicular tumours. Shortly after, an English-speaking group started its activities in Yorkshire, with main emphasis on prostate cancer, and a "bladder cancer group" attracted many urologists from Belgium and other European countries. In 1976 all these groups were fused into a one new urological group of which M. Pavone-Macaluso from Palermo, Italy, secretary of the French-speaking group, was chosen as secretary and then chairman of the unified group, whereas Ph. Smith from Leeds, UK, chairman of the English-speaking group, was elected as chairman and later as secretary of the new group. The two "founding fathers" celebrated the 10th and 20th anniversaries of the foundation of the group respectively in Leeds in 1986 and in Palermo in 1996. Later chairman were L. Denis, Belgium; F. Schroder, The Netherlands; D. Newling, UK; F. Debruyne, The Netherlands and R. Hall, UK. A. van der Meijden, The Netherlands, will take over in 1997. The present secretary is A.V. Bono from Varese, Italy. The present structure of the group consists of a variety of working parties (disease orientated groups), with special interest in a given pathology (such as prostate cancer, superficial or advanced bladder cancer, etc.) and of other committees (chemotherapy, quality of life, quality control). All the activities are coordinated by a Data Centre in Brussels, that is responsible for the statistical support. In its 20 years of activity the group has made many contributions of significance which have coincided with a number of changes in the urological oncology and have obtained international recognition. The paper analyses in detail the most significant of the group's achievements in the various fields of urological oncology. PMID- 9417294 TI - [Wilms' tumor: reached progress and future prospects]. AB - Wilms tumor, although rare, is the most frequent malignant renal tumor in children. With approximately 70 new cases diagnosed annually in Italy, it is important to collect these rare cases and to treat them in a uniform way. At the dawn of this century virtually all children with Wilms tumors died: today 95% of them survive; the treatment is carefully tailored to well defined risk factors (histology-stage-molecular markers) to minimize short and long-term toxicities. The object of this review is to describe the pathway to the progress. The most important step was to recognize that a single institution could not collect enough patients to answer complex questions, whereas this was possible for super groups like the National Wilms Tumor Study Group (NWTSG) in the USA, the multinational SIOP study group in Europe, the United Kingdom group, the Italian group (CNR-AIEOP) and more recently the Brazilian one. Wilms tumor is the paradigm for the multimodal treatment of a pediatric malignant solid tumor, the development in surgical technique, the recognition of the sensitivity of Wilms tumor to irradiation and the availability of several chemotherapeutic agents led to a dramatic change in the prognosis for most patients. Much progress has been made in the study of histopatology of childhood renal tumors: the patients must be stratified into two groups, the favourable and the unfavourable histology, further subdivided into anaplasic tumors (focal or diffuse), clear cell tumors and rhabdoid tumors. The past few years have provided a breakthrough in understanding some of the genetic factors involved in Wilms' tumor: moreover, possible chromosomal prognostic factors have been identified: loss of heterozygosity of 16q markers and 1p markers. Today the results of the treatment of Wilms tumors are very good. In the NWTS-3 the four year relapse-free survival rate in stage 1 with favourable and with anaplastic histology was 92%, in stage 2 with favourable histology 88%, in stage 3 with favourable histology 79%, in stage 4 and in stage 2, 3 and 4 with unfavourable histology 71%. The Italian group has obtained less impressive results in the ?80, but similar results in the first stage with the ?92 protocol. There is a debate about the immediate nephrectomy preferred by NWTS and the preoperative strategies adopted by SIOP group. Successful treatment may be associated with many late effect: in patients cured of Wilms tumor the risk of congestive heart failure has been less than 1.7%, the risk of a second tumor less than 1%. The must important late effect remains the relapse of the disease: the risk is about 14-20%. PMID- 9417295 TI - [Recent advances on retroperitoneal neuroblastoma]. AB - Neuroblastoma, a malignant tumor of infancy and childhood, has some very interesting peculiars: good prognosis, even with disseminated disease, propensity to occasionally undergo spontaneous regression, its ability to undergo spontaneous or induced differentiation to a benign ganglioneuroma. Neuroblastoma may originate anywhere along the sympathetic nervous system chain. The most common site of primary tumor is, however, within the abdomen either in the adrenal gland or in a paraspinal ganglions. A great deal of progress has been made in advancing the knowledge of human neuroblastoma at the cellular and molecular viewpoint. The genetic predisposition to develop the tumor is clarified, a specific oncogene amplified (N-myc) in neuroblastoma cells shows precise prognostic significance and the deletion of chromosome 1's short arm has been defined. Work-up in neuroblastoma's diagnosis include the urine assay for catecholamine metabolites (VMA, HAVA, VLA) and serum assay for the specific markers as neuron-specific enolase (NSE), ferritin, GD2 ganglioside. Imaging include CT-scan, MIGB body-scan and the newest monoclonal antibodies scan. Abdominal tumors are shown in about 75% of children > 12 months old. In 2/3 of cases, tumor is widely disseminated at the time of diagnosis. In the period 1979 94 the Italian Group for Neuroblastoma (GCN-AIEOP) collected 1083 cases of tumors and 5-yrs survival was 45% +/- 2.4 for the patients studied in the period 1979 84, which is increased to 58% +/- 3 for the group of patients 1990-94 (p < 0.001). The overall survival was 53 +/- 1.7. About 5-yrs survival at different stages, AIEOP shows that it is increased from 88% +/- 3.3 (1979-84 group) to 91% +/- 2.8 (1985-92) in the stage I and II (280 cases). In the stage IV survival value improved from 79% +/- 7.1 to 84% +/- 7 (132 cases). No statistical improvement can be observed, anyway. Better improvements can be pointed out in stage III (221 cases, survival from 48% +/- 5.2 (79-84 group) to 69% +/- 4.8 (85 92) and stage IV (483 cases, survival from 16% +/- 2.6 to 28% +/- 3.4) (p < 0.001). Finally we can summarize about neuroblastoma: 1) better prognosis in the first year of life; 2) ability to spontaneous regression, first of all, in stage IVs; 3) partial and provisional response to therapy in advanced stages; 4) no recovery increasing despite advancing in surgery and chemotherapy. PMID- 9417296 TI - [Heredity in renal and prostatic neoplasia]. AB - There is an ever growing report of data supporting the evidence that accumulated genetic changes underlie the development of neoplasia. The paradigma of this multistep process is colon cancer were cancer onset is associated, over decades, with at least seven genetic events. The number of genetic alterations increases moving from adenomatous lesions to colon cancer and, although the genetic alterations occur according to a preferred sequence, the total accumulation of changes rather than their sequential order is responsible of tumor biological behavior. It is noteworthy that, at least for this neoplasia, carcinogenesis appears to arise as a result of the mutational activation of oncogenes coupled with the mutational inactivation of tumor suppressor genes. In some cases mutant suppressor genes appear to exert a phenotypic effect even when present in the heterozygous state thus been non "recessive" at the cellular level. The general features of this model may apply also to renal cell cancer (RCC) and prostate cancer (CaP). Extensive literature exists on the cytogenetic and molecular findings in RCC. Only 2% of RCC are familiar, but molecular genetic studies of these cancers have provided important informations on RCC pathogenesis. As with other cancers, familiar RCC is characterized by an early age of onset and frequent multicentricity. A pathological classification useful in studying these patients subdivide renal cancers in papillary (pRCC) and non papillary (RCC) neoplasms. The most common cause of inherited RCC is the Von Hippel Lindau disease (VHL) a dominantly inherited multisystem disorder characterized by retinal and cerebellar hemangioblastomas, pheochromocytomas, pancreatic cysts and RCC. Over 70% of these patients will develop an RCC by their sixth decade. In 1993 the isolation of the tumor suppressor gene in VHL disease at the level of chromosome 3p25-p26 have lead to a better understanding of RCC. Most missense mutations are associated with high risk of RCC, but some are associated with high risk of pheochromocytoma and low risk of RCC. The VHL gene is evolutionary conserved and encodes for a specific protein (pVHL). VHL protein downregulates transcriptional elongation and so suppresses the expression of proto-oncogenes and growth factors. Recently reintroduction of wild-type, non mutant, VHL gene into VHL deficient RCC cell line 786-O had no demonstrable effect on their in vitro growth but inhibited their ability to form tumors in nude mice. So far, VHL mutations or hypermethylations have been found in 76% of sporadic RCC. On the contrary, up to now, no 3p allele loss or VHL mutations have been detected in pRCC. Preliminary studies in familiar pRCC are pointing on genetic changes on chromosomes 1, 7, 16 and 17. As far as prostate cancer is regarded, men with a family history of prostate cancer have an age dependent, significantly increased PCa risk. For familiar clustering, of PCa the two main factors are early age at onset of the disease and the number of multiple affected family members. Hereditary prostate cancer is a subset of familiar prostate cancer with a pattern of distribution consistent with Mendelian inheritance. Hereditary prostate cancer is clinically defined as a clustering of 3 or more relatives within any nuclear family; or the occurrence of prostate cancer in each of 3 generations in either the probands paternal or maternal lineage; or a cluster of 2 relatives affected within 55 years of age or less. Therefore, hereditary prostate cancer may be seen as a multistep carcinogenesis, and clustering may be explained by Mendelian inheritance of a rare (frequency in population 0.36%) dominant, highly penetrant, allele. The estimated cumulative risk of developing PCa, is 88% for carriers as compared with 5% for non carriers. There are conflicting reports of an associated increased incidence of breast cancer in female relatives of men with familiar prostate cancer. In conclusion, there is a clear associatio PMID- 9417297 TI - [Hormone receptors and growth factors in carcinoma of the prostate]. AB - The therapeutic and prognostic significance of androgen receptors in prostatic carcinoma has been examined on the basis of data obtained with the different techniques used in receptor determination. Moreover, the role of some polypeptide growth factors in the regulation of prostatic cancer growth and progression has been reviewed. Great attention has been focused on in vitro models utilized to investigate androgen receptor alterations and the effects of the different positive and negative regulators of prostatic carcinoma cell growth. PMID- 9417298 TI - [Molecular biology in bladder carcinoma: contributions of immunohistochemistry]. AB - Despite the insights genetics and molecular biology have given to a better understanding of the mechanisms which lead to the onset and development of bladder carcinoma, the factors that influence its unpredictable and, at times, particularly aggressive outcome are still largely unknown. Also in bladder carcinoma the study of cellular differentiation markers has been replaced by that of genotypic alterations, and, mainly with the help of immunohistochemistry, of the expression of genes involved in cell proliferation and death, such as MTS1, TP53, Rb, c-myc, Bcl-2, c-erb-B2. So far, anyway, no independent and reliable indicator able to predict the outcome of the single tumour has been identified, and this issue seems to be best addressed by studies of the altered expression of more than one oncoprotein simultaneously. Fairly identical is the question arised by TP53 mutations, which, while worsening the evolution of advanced muscle infiltrating tumours, hold a still unclear and debated meaning in superficial tumours. It is anyway clear that molecular analysis only may enable to reliably detect the presence of any TP53 mutations. As a matter of fact, the multiplicity of genetic mutations, the frequent transcript variations and the intrinsic limits of immunohistochemistry may explain the discrepancy between immunohistochemical and molecular analysis results, with specificity and sensitivity levels clinically not acceptable. To date, anyway, the biological and clinical meaning of this discrepancy has still to be clarified, as well as the clinical meaning, if any, of p53 overexpression in the absence of gene mutations. PMID- 9417299 TI - [Cellular proliferation, expression of p53, EGFR and apoptosis index of healthy mucosa of the bladder with TCC; pre- and post-intravesical BCG immunohistochemical study]. AB - At present, the most efficacious and used immunostimulant agent in the superficial bladder cancer immunotherapy field, is the BCG, even if its mechanism of action is still partly unknown. The therapeutic effects of BCG don't seem to depend exclusively on local immune response, so that according to this assertion, this immunohistochemical study had been conducted on 14 patients affected by superficial bladder cancer (pTa-pT1) which aimed to value both the apoptosis and proliferation indexes and the expression of the genetic product p53 and EGFR before and after the exposition of the vesical mucosa to the BCG. The BCG treatment can reduce the proliferation index of the normal urothelial cells in a statistically significant way whereas it would exclude a cytostatic effect mediate by negative modulation of EGFR from the cytokinins induced by BCG itself. The index of apoptosis of the urothelium does not increase after BCG and decreased expression of p53 associated after the treatment, although statistically not significant, it would seem to bear, the prophylactic efficacy of BCG according to the follow up of the patients included in the study. PMID- 9417301 TI - [Effects of previous small-dose irradiation on blood reaction and survival of mice after subsequent radiation and combined radiation-thermal injuries]. AB - CBA x C57BL6 mice pre-exposed to 5 cGy gamma-irradiation then inflicted to LD70/30 acute radiation or combined injuries (radiation + thermal burn). In such protocol of the experiments preliminary "adaptive" dose radiation did not modify low 30-day survival rate and did not render an influence to the mean survival time. Leucopenia level and bone marrow devastation in 3 days after sublethal irradiation (4 Gy) or combined injury was similar in mice pre-exposed to low dose radiation (5 cGy) and in their unexposed controls. "Adaptive" dose radiation decreased leukocytes' number within the phase of hemopoiesis recovery (in 14 day after sublethal irradiation or combined injury). PMID- 9417300 TI - [Distribution of ionizing radiation absorption in microstructures of a biosystem]. AB - The method of determination of the gamma-radiation absorption distribution in microstructures of a biosystem is proposed. The absorption capacity and energy absorption distribution of some biomolecules, mass coefficients and optical density of blood and thyroid gland are calculated. Influence of the pollution of biosystem microstructures (liver, blood, thyroid, gland) by hard elements on the energy absorption distribution is demonstrated. PMID- 9417302 TI - [Radiation-protective effect of agonists of alpha-2 adrenergic receptors. Methyldopa]. AB - MethylDOPA has the considerable (60-80%) and lasting RPE at both intraperitoneal (0.5-3.8 mmol/kg) and per os (3.8-7.1 mmol/kg) introduction in radiation dose 8 Gr (LD97/30). Optimal time for introduction is 0.5-3.0 hours intraperitoneally and 3-6 hours per os. Evidently, methylDOPA RPE is realized via alpha 2 adrenoceptors. PMID- 9417303 TI - [Biological preparation based on Lactobacillus acidophilus, a new agent in the early treatment of combined radiation-thermal injuries]. AB - Male mice F1 (CBA x C57BL6) were used for experiments. Animals were exposed to 7 Gy gamma-radiation and additionally inflicted to full-thickness thermal burn 10% body surface. As it has been revealed, single subcutaneous injection of the created biopreparation based on inactivated lactobacillus microbic biomass increased mice survival with combined injury from 23% to 73%. Therapeutic efficacy of this remedy did not correlate with postradiation damages of the hemopoietic system. Injection of killed L. acidophilus mixture strikingly averted development of the intestine autoinfection. Possible mechanisms of the benefit therapeutic action of the new preparation are discussed. PMID- 9417305 TI - [Behavior reactions and lipids of brain synaptic membranes of rats under chronic exposure to gamma irradiation]. AB - The effects of low level chronic ionising irradiation (12.9 cGy/day on the sensory attention to the stimuli of different modalities (somatosensory, visual, odor) of Wistar rats were studied. Analysis of animals behaviour was made after they had received the different doses of irradiation: 4, 6, 8, 10, 15 and 20 Gy. It was founded, that the attention and exploratory activity of rats is significantly decreased up to 20-30% after 4-6 Gy. The irradiation doses 8 Gy did not change animal behaviour as compared to control animals, but doses 10, 15 and 20 Gy decreased the exploratory activity as well as sensory attention of rats to 3-5-times as compared to previous dose. Such a wave-like way of behaviour reflects the functioning of an adaptive mechanism. Biochemical data indicated that after 5 months of the irradiation (dose 20 Gy) the level of phospholipids, lysophosphatidylcholine, phosphatidylethanolamine, phosphatidylcholine, cholesterol were decreased. PMID- 9417304 TI - [Biological effects of acute external or internal irradiation of rats in the framework of nitrite intoxication]. AB - The material presented provides evidence that high nitrite doses have a various effect under the condition of external or internal radiation. Thus, nitrite protector properties were observed in male rats exposed to external radiation using the LD50/30 while following 90Sr exposure male rats of the same age showed a higher sensitivity to the radionuclide. The nitrite combined with optimum carcinogenic 90Sr dose had a potentiating effect on both life-span shortening and increase in osteosarcoma rate in the animals. PMID- 9417306 TI - [Vascular permeability of an inflammatory focus in irradiated animals]. PMID- 9417307 TI - [Functional state of the supraepithelial mucous layer of the digestive tract in mice in the postnatal period after irradiation]. AB - It was studied the condition of mucous layer of intestine in mice model experiments after ionizing radiation. It were found out the differences in reaction of intestine on radiation in mice at the age of 55 and 95 days. In mice at the age of 55 days at the whole stretch of intestine was marked increased secretion of the polymeric glycoproteins, whereas in mice at the age of 95 days such alterations were marked at smaller degree. In both groups mice was marked increased secretion of bicarbonate after radiation, that is possible to count as characteristic compensative mechanism. PMID- 9417308 TI - [Contents of general, bound and free cortisol in sheep blood during different periods of acute radiation sickness]. AB - The hypothalamic-hypophyseal-cortical multifetal sheep system reaction was examined to respond the sharp single action X-rays dosed 0.96 and 3.84 Gr. The primary irradiation answer was accompanied by expressed hypercorticoidism, which essentially increased with irradiation dose rising and followed the induction of CRF-hypothalamic activity. The secondary sheep hypercorticoidism preceded the clinico-hematomancy and depended on irradiation dose. Hypercorticoidism-2 resulted in blood free bioactive cortisol content increasing. The compensative decrease of named system activity favoured to beam illness outcome. It the prognosis was poor the free and reached its peek before animal death. These data can be used as a test for beam illness outcome prognosis. PMID- 9417309 TI - [Health status of persons with acute radiation sickness caused by Chernobyl AES accident]. AB - After Chernobyl accident 75 survivors with acute radiation syndrome (ARS) have been under observation in the Clinic of the Institute of Biophysics. Years after accident serious problems occurred in the individuals, who suffered from late effects from radiation injuries (late radiation ulcers and radiation cataracts). One patient suffered on hypernephroma, one-kidney cyst. Observed somatic diseases are gastrointestinal diseases, different neurological diseases and syndromes and upper pulmonary tract diseases. Hemopoiesis system condition was characterised by the development of transient moderate cytopenia (light and moderate degree of ARS patients) as well as the periodical increase of blood cell counts above standard indices (severe degree of ARS patients). PMID- 9417310 TI - [Behavior of fuel hot particles in the body of cows at oral intake]. AB - It was studied the behaviour of fuel hot particles (analogous to Chernobyl) in gastrointestinal tract of cows. The values of caesium and strontium radionuclides transfer to the cows organism and its transition parameters to milk after the single per oral intake to the organism of animals are estimated. It is shown, that the biological simplicity of radionuclides in the fuel hot particles at two parameters lower, than the same radionuclides in washed phases. PMID- 9417311 TI - [Methods of analysis of protective measures in agriculture regarding radioactively contaminated land: evaluation of the effectiveness, intervention levels and comparison of different countermeasures]. AB - Methodological aspects of the analysis of protective measures in agriculture in the long-term period of liquidation of the consequences of a nuclear accident are considered. Examples of the estimations of countermeasure effectiveness with the use of the cost-benefit analysis, as well as methods of the estimation of intervention levels and examples of a comparison of various protective measures with the use of several criteria of effectiveness are discussed. PMID- 9417313 TI - [Possibilities of microtomography of blood cells]. AB - Social reforms and the technological revolution lead to changes in the environment and human habitat, which deteriorate the ecology in the majority of cases. Blood system in general, specifically, the peripheral blood represent one most sensitive system of the body, reflecting the level of adverse exposures and reaction of the organism to these exposures. The interference microtomographer permits measurements of the spatial structure of optically transparent three dimensional cytological specimens (blood cells). During preliminary studies a model of interference computer-aided microtomographer has been created and a method for treating preparations for processing by this microtomographer developed. PMID- 9417312 TI - [Use of multivariate methods in the analysis of laboratory indicators of blood]. AB - Cluster and discriminant methods were used for analysis of the time course of 12 routine biochemical characteristics of murine serum after tumor transplantation. Mathematical methods helped single out groups of animals with the same prognosis. The methods were tried in clinical studies. PMID- 9417314 TI - [New possibilities of optic microscopy in studies of blood cells]. AB - Modern biomedical tasks put forward new requirements to the instrumental base of microscopy. This paper outlines the prospective trends in microscopy which are the most interesting for medicine, on the one hand, and analyzes the probability of meeting the requirements of medicine by modern methods of optic electronics and computer processing of optic information. The authors analyze the following trends: microscopy of three-dimensional objects, super-resolution, microscopic measurements, and automation of microscopes. Progress in optic electronics and computer processing of images permits considering these trends as a priority task of modern microscopy. PMID- 9417315 TI - [Device for assessing the electrophoretic mobility of erythrocytes and other corpuscles in dispersive systems]. PMID- 9417316 TI - [Experience in the use of flow scintillator Cobas Micros-18 OT in hematological practice]. AB - Clinical analysis of the blood is a most prevalent investigation in practical medicine. The need in effective assessment of the results of blood analysis prompted the creation of automated flow systems. Our task was to analyze the blood of normal subjects and patients by Hoffmann La Roche Cobas Micros-18 OT (France). It is a completely automated hematological analyzer used for in vitro diagnostic tests of whole blood. The work consisted of two steps: 1) comparison of the results of leukocyte and red cell counting by Cobas Micros and Goryayev chamber and 2) assessment of the leukocyte formula in blood donors and patients with diseases other than hematological by the Cobas Micros device and compare the results with the data of visual assessment in optic microscope. The results of analysis by the automated scintillator, Goryayev's chamber, and optic microscopy were compatible. Cobas Micros is very convenient for analysis of liquid blood on an outpatient and inpatient basis and during overall screenings. The device facilitates the deciphering of the leukocytic formula and assessment of the status of formed elements of the blood in mass screenings and detection of abnormalities. PMID- 9417317 TI - [Principles of creating software and equipment for automating the workplace of laboratory physician]. AB - Automation of laboratory tests is a pressing problem of today, for examinations of the hemopoietic system is a priority in the diagnosis of the majority of diseases and in detecting the effects of unfavorable ecological factors on man; in addition, laboratory studies are time- and labor-consuming. A system "Automated Work Place of a Laboratory Physician" has been developed, which permits automated scanning of a routinely stained blood smear with automated focussing on the detected objects. A preset number of objects is filed in the archive for further identification by a morphologist. The results can be stored in the information database for each patient and be transferred through telecommunication network. The system includes a computer hypertext Blood Atlas. PMID- 9417318 TI - [Study of the peripheral blood of mice using automated analyzer Cobas Micros in experimental animals]. AB - Automated analyzer intended for analysis of human blood assesses the main parameters of murine blood with high accuracy and good reproducibility. Differentiation of murine leukocyte by this analyzer is objectively impossible, but it permits the investigation of some characteristics heretofore difficult to study because the routine methods were inaccurate (assessment of the mean size of red cells and platelets or of the mean content of hemoglobin per red cell). Analysis of these values is useful, among other things, in simulation of various anemias in mice. PMID- 9417319 TI - [Laboratory assessment of the renal function (a lecture)]. PMID- 9417320 TI - [Flow cytometry of bone marrow cells in health, anemias of different etiology and acute leukemias]. AB - The distribution of bone marrow cells by stages of cell cycle was studied by flow cytometry in 26 normal subjects and 107 adult patients (42 with anemias and 65 with acute lymphoblastic or myeloblastic leukemias). Normal myelokaryocyte cell cycle is rather stable, which manifests by low variability of cells. In anemias and, more so, in leukemias the proliferative status of cells varies within a wide range, which is explained by unstable and ineffective hemopoiesis. PMID- 9417321 TI - [Computer-assisted microtelephotometric study of dysplasia and adenocarcinoma of the large intestine in patients with chronic nonspecific colitis]. AB - Computer-aided microtelephotometry showed that the mean size of the nuclei gradually increases and their shape changes, and the total content of Feulgen DNA increases in patients with nonspecific ulcerative colitis in proportion to the severity of dysplasia and malignant degeneration of the mucosal epithelium of the large intestine. In first-degree dysplasia the value os 2.8, in second degree 5.0, in third degree 6.9; in cribriform structures of adenocarcinoma 8.6, in poorly differentiated cancer cells 10.9, and in highly polymorphous sites of adenocarcinoma as high as 20. Mathematical expression of different degree of dysplasia and of adenocarcinoma of the large intestine will no doubt facilitate the differential diagnosis of these processes and help universally assess them. A new technology of histocytological investigations - computer-aided microtelephotometry - permits the use of all types of telecommunications for consultations to be carried out all over the country. PMID- 9417322 TI - [Large-cell anaplastic lymphoma Ki-I]. PMID- 9417323 TI - [In vitro prognostication of individual chemosensitivity of surgically removed and biopsied tumors]. PMID- 9417324 TI - [Differential diagnostic and prognostic significance of the activity of nucleolar organizer region in non-Hodgkin's malignant lymphoma]. PMID- 9417325 TI - [A standard cytological conclusion]. PMID- 9417326 TI - [Possibilities of the use of tumor markers in specifying the severity of ulcerative colitis]. PMID- 9417327 TI - [Automated cytophotomorphometric tests of blood smears in general clinical studies and population screening]. AB - Automation of routine operations of photomorphometry of blood smears realized in the MEKOS-C cytoanalyzer helped create a new level of using a set of cytological tests, such as assessment of the leukocyte and erythrocyte formulae and of the histogram of hemoglobin distribution in red cells, estimation of leukocyte subtypes, assessment of the characteristics of the nucleolar system and the nucleoli-forming sites. Automated complex analysis of an increase in the measured characteristics of red and white blood cells impossible in common microscopy and flow cytoanalyzers appreciably improves the sensitivity and accuracy of tests and extends the range of their application. Low cost of analysis of a blood smear permits the use of MEKOS-C not only in clinical studies, but even in overall prophylactic screenings of the population. PMID- 9417329 TI - [Transfusion Medicine 1996/7. Proceedings of the 29th Congress of the German Society of Transfusion Medicine and Immunohematology. Essen, 11-14 September 1996]. PMID- 9417328 TI - [Analysis of erythrocytes in the MEKOS-C system]. AB - Quantitative cytofluorimetric analysis of red cells by the MEKOS-C device provides complete information about peripheral blood erythrocytes, which cannot be obtained by other methods and equipment. An analysis is cheap and sensitive, this recommending it for practical hematology, toxicology, and narcology. The MEKOS-C device permits the preclinical diagnosis of a number of diseases and monitoring their treatment. The device is cheap and available for therapeutic and diagnostic institutions of all levels. PMID- 9417330 TI - [Study by the Professional Organization of German Transfusion Physicians on epidemiology of HIV- and hepatitis infections in blood donors]. AB - The frequency of the seromarkers of HIV, HCV, and HBV in the donor population from all donor centers in Germany became available from the multicenter study of the transfusion services. From these data the risk of virus transmission by blood products can be estimated at 1:1 million for HIV and 1:50,000 for HBV. For HCV, preliminary data indicate the risk for Germany in less than 1:20,000. The risk is even lower, with less than 1:200,000 when calculated from seroconversion data of a rural donor population. The safety of blood products has been very much improved due to the continuous donor selection over the last years and by the epidemiologic situation in Germany. PMID- 9417331 TI - [Cytokines and chemokines as inducers of nonhemolytic transfusion reactions after thrombocyte transfusion]. AB - Recently, the leukocyte-derived cytokines interleukin (IL)-1 beta, IL-6, IL-8, and tumor necrosis factor(TNF)-alpha were found in considerable concentrations in platelet concentrates (PC) involved in nonhemolytic transfusion reactions (NHTR). We investigated transfusion reactions in two study periods over the last 3 years. In the 1st period, PC were leukocyte-depleted by bedside filtration. In the 2nd period we performed prestorage leukocyte filtration of PC. Recipients who experienced a transfusion reaction were tested for total and specific IgE, and reactions were analyzed with regard to the main symptoms. In the supernatant of platelets involved, we analyzed concentrations of IL-1 beta, IL-6, IL-8, TNF alpha, macrophage inflammatory protein(MIP)-1 alpha, and RANTES. The incidence for NHTR did not change with the kind of PC transfused (n = 44, 1.63% and n = 46, 1.56%; p = 0.84), but allergic reactions were reduced in the 2nd observation period (0.92% versus 0.51%; p = 0.02). IL-1 beta, IL-6, IL-8, TNF-alpha, and MIP 1 alpha were not detectable in the majority of all products. PC involved in allergic reactions contained significantly higher concentrations of RANTES. We found evidence that the proinflammatory chemokine RANTES, stored in the platelet alpha-granules, is involved in allergic transfusion reactions. In contrast, leukocyte-derived inflammatory cytokines were of minor relevance in the onset of NHTR. PMID- 9417332 TI - [Prevalence of hepatitis G virus genome in blood donors]. AB - The relevance of the GB virus C/hepatitis G virus (GBV-C/HGV) in blood banking results from its high prevalence in blood donors and the fact that it is present at a very high percentage (18%) in polytransfused and hemophiliac patients. Since there is no assay available for serological testing, we developed a sensitive PCR utilizing newly designed NS3 primers to investigate the prevalence in blood donors in Hessia. Testing 1,143 accepted blood donors with alaninaminotransferase (ALT) concentrations < 45 U/l we found 15 (1.3%) positive for GBV-C/HGV RNA. From 507 donors settling in urban Frankfurt areas, 10 (2.0%) were positive. Of those donors settling in rural hessian areas only 5/635 (0.8%) tested positive. By testing 100 excluded donors with ALT values > 45 U/l, 3% turned out to be positive. In 4 out of 9 recipients of GBV-C/HGV-positive blood products we detected the donor viral RNA as proved by sequencing and phylogenetic analysis. The virus was transmitted to 3 recipients by erythrocytes and to 1 recipient by platelets. Testing family members of the GBV-C/HGV-positive blood donors we could not detect any intrafamilial transmission. PMID- 9417333 TI - [In-line leukocyte depletion ov thrombocytapheresis concentrates with the Fresenius-AS-104 cell separator]. AB - This study reports on in-line filtration of 72 platelet concentrates (PC) collected by the Fresenius AS 104 cell separator, using the new C4F sets with integrated leukocyte filters (Biofil P plus). MATERIAL AND METHODS: 72 volunteer donors, automatic counts of platelets, microscopical counting of residual leukocytes with the Nageotte chamber, GMP-140 by flow cytometrie, beta thromboglobulin release, platelet aggregation (ADP, collagen). RESULTS AND CONCLUSION: Filtration reduced leukocytes by 98.5%. Residual leukocyte contamination remained clearly below 5 x 10(6) (mean 0.5 +/- 0.6 x 10(6), maximum 2.8 x 10(6). Platelet loss by filtration was found to be between 27.4 and 0.7% (median 8.5%). Filtration caused a significant decrease of platelet aggregability (p < 0.005), but no significant increase of beta-thromboglobulin release and only a slight decrease of GMP-140 expression. From these data can be concluded that in line filtration was highly efficient with acceptable platelet retention. No significant platelet activation could be observed in the PC. The decrease of platelet aggregability have been due to the reduction of activated platelets which are believed to show reduced in vivo survival. PMID- 9417334 TI - [Preparation of leukocyte depleted thrombocytapheresis preparations using closed disposable systems with an in-line filter for the AS 104 cell separator]. AB - Plateletpheresis kit with an integrated leukocyte filter of three different batches were evaluated in the present study. White cell content after leukocyte depletion was in the range of 10(4)/unit. Modification of the filtering matter density and thickness resulted in a platelet recovery > 85%. PMID- 9417335 TI - [A new separation protocol (DRBCP-F) for automated blood component donation with the MCS 3p cell separator for collection of leukocyte depleted erythrocyte concentrates and plasma]. AB - Previously published studies on automated blood component donation with the MCS 3p cell separator proved fairly good quality of the collected red blood cells (RBC) and fresh frozen plasma (FFP), with the disadvantage of a low hematocrit of the filtered RBC and a high platelet contamination of the FFP (RBCP-F protocol.) The DRBCP-F protocol was designed to eliminate the above-mentioned disadvantages and to provide 1 unit of leuko-depleted (filtered) RBC, 2 units of FFP, and additionally 1 platelet concentrate (PC) from the buffy coat. Twenty automated blood component collections (2 cycles, Latham bowl at 5,500 rpm, 230 ml isotonic saline for volume balance, PAGGS-M as additive solution) were performed. The RBC were filtered in a closed system after storage at 4 degrees C for 24 h. Blood cell counts and biochemical parameters of the RBC were determined initially and after 49 days. PC were separated from buffy coat after a soft spin. The volume of the RBC amounted to 293 +/- 12 ml (mean +/- SD) with a hematocrit of 0.61 +/- 0.05 l/l. Residual leukocytes after filtration were found to be 0.04 x 10(6) +/- 0.06 per unit. After storage, the following data were obtained: hemolysis 0.38%, ATP 2.1 +/- 0.4 mumol/g Hb, 2,3-diphosphoglycerate (2,3-DPG) 1.4 +/- 0.3 mumol/g Hb, ph 6.3 +/- 0.1, potassium 6.4 mmol per unit, and LDH in the supernatant was 219 U/l. None of the RBC showed bacterial growth after 49 days. The volume of the collected FFP was 398 +/- 32 ml, with 3.4 +/- 3.5 x 10(3) residual platelets and 5 +/- 12 leukocytes per microliter. Platelet concentrates contained 90.2 +/- 32 x 10(9) platelets in 88 +/- 14 ml plasma. Automated blood donation with the DRBCP-F protocol provided RBC with very low residual leukocyte counts, adequate hematocrit and good metabolic status up to 49 days, and FFP with low platelet contamination. The platelet concentrates were even superior to those prepared from whole blood using the buffy coat method. The storable leuko-depleted RBC are suitable for transfusion of chronically transfused patients in whom primary HLA sensitization should be prevented. PMID- 9417336 TI - [Changes in blood rheology and microcirculation after preparatory combined thrombocytapheresis and plasmapheresis]. AB - INTRODUCTION: Because of anticoagulation, changes in blood composition, and- perhaps--extracorporeal circulation, donor apheresis should cause alterations in hemorheology and hence in the perfusion of the microvasculature. MATERIAL AND METHODS: 19 regular blood donors were included. According to our standard protocol for automated collection of blood components with the MCS 3p cell separator, we harvested 1 unit of platelets and 2 units of plasma in each case. Prior to, 1 h after, and 24 h after donation, the following parameters were measured: total serum protein (tsp), hematocrit (hc), whole blood and plasma viscosity (wbv/pv), red cell aggregability (rca) and blood flow velocity of the nail-fold capillaries (bfv). RESULTS: The following parameters decreased 1 h/24 h after donation: tsp (p < 0.001/p = 0.008), elastic wbv (p = 0.018/p < 0.001), viscous wbv (p = 0.85/p = 0.0031), pv (p < 0.001/p < 0.001), static rca (p < 0.001/p = 0.0073), dynamic rca (p < 0.001/p = 0.017). The hc showed an initial increase (p < 0.001) with a subsequent overshooting decrease after 24 h (p < 0.001). 1 h after donation bfv raised (p = 0.0065). It decreased after 24 h and remained only slightly higher than the initial level (p = 0.27). CONCLUSIONS: Automated combined collection of platelets and plasma gives rise to: i) Improvement of rheological properties of the donor's blood and increased bfv of his nail-fold capillaries within the 1st h after apheresis. ii) 24 h after donation the improved hemorheological properties remain demonstrable, but the bfv of nail-fold capillaries declines and shows a trend toward the starting-point. iii) Taken together, this is possibly reflecting adapted hemodynamic and vasoconstrictor regulation for altered hemorheological conditions. PMID- 9417337 TI - [Effect of processed blood volume, leukocyte count and concentration of CD34 positive cells in peripheral blood on efficiency of stem cell apheresis]. AB - Despite many published studies no parameter could be identified yet to acceptably and individually predict collection results in stem cell apheresis. We analyzed leukocyte counts and processed blood volume, absolute and relative CD34+ cell counts, and overall collection efficiency in 120 patients with hematological and solid malignancies (354 leukaphereses using the Cobe Spectra cell separator, a median of 3 per patient, span 1-9). Stem cells were mobilized into peripheral blood by conventional chemotherapy followed by daily doses of G-CSF. CD34+ progenitor cell counts were monitored through multiparametric flow cytometry. Blood and collection flows varied in the range of 45-90 ml/min and 0.7-1.5 ml/min, respectively. CD34+ progenitor cells were enriched 38-fold in the apheresis product as compared to peripheral blood at a processed blood volume lower than one total blood volume. Efficiency continuously declined, on to a 25 fold concentration at a processed blood volume above the 3-fold total blood volume. Total collection efficiency, calculated from the absolute content of CD34+ progenitor cells in peripheral blood and apheresis concentrate (a parameter for progenitor cell mobilization during the apheresis), reached a plateau at a processed blood volume above the 3-fold total blood volume. However, variation among individual patients was high. The concentration rate of CD34+ cells at a leukocyte count below 5,000/microliter averaged 50 and declined continuously to 8 at leukocyte counts between 45,000 and 50,000/microliter. To summarize, in 70% of patients with leukocyte counts below 5,000/microliter and CD34+ progenitor cell counts above 10,000/ml, more than 1.5 x 10(6) progenitors per kg body weight could be collected in a single leukapheresis. According to the presented data, the variation in overall collection efficiency is mainly due to: 1) varying mobilization of progenitors during the apheresis procedures itself and 2) dependence on peripheral leukocyte counts. PMID- 9417338 TI - [Effect of rhG-CSF on platelets using in vitro bleeding time and transcranial Doppler examination]. AB - There is experimental evidence for recombinant human granulocyte colony stimulating factor (rhG-CSF)-induced stimulation of hemostaseologic parameters including platelets. Using the closure time of the in vitro bleeding test (IVBT, Thrombostat 4000, VDG, Seeon) and different hemostaseologic parameters, our results are suggestive of a directly or indirectly induced in vivo stimulation of platelets following rhG-CSF stimulation in healthy stem cell donors. According to the negative transcranial Doppler microembolic monitoring of the middle cerebral artery, there is no evidence for clinically silent cerebral microembolism supporting a prothrombotic state facilitating the formation of arterial thrombi with subsequent cerebral embolization. PMID- 9417339 TI - [Capture-R Ready-ID and DiaMed-ID for identification of erythrocyte bound antibodies after acid elution]. AB - The DiaMed-ID (D-ID) gel system is known to be a very sensitive and specific method for the detection of red cell antibodies. In various cases, we failed to find an antibody in the eluate in which red-cell-bound antibodies (IgG) where proven by a positive direct antiglobulin test (DAT). SPRCA (Capture-R, Ready Screen, Immucor; C-R) seems also to be very sensitive, in part due to the antihuman, IgG-coated indicator cells. Therefore, we compared 39 acid eluates from patients who had a positive DAT (monospecific rabbit antihuman IgG) both in the D-ID and in the C-R system. Patients (19 female, 20 male; mean age 62 years) were suspected either to have an autoimmune hemolytic anemia or an alloantibody. Identification of the antibodies was done with the system's own panel cells. Agglutination strength was scored from 1 to 4. Quantification of selected eluates was performed by titration, using the same cells in both systems. From 39 eluates, 31 were positive in the C-R and 25 in the D-ID. Nine eluates were negative in the C-R and 14 in the D-ID. If only eluates with a DAT reaction strength of 2 or lower were considered, obviously more negative results were found with the D-ID technique (p < 0.027) than with the C-R technique. In all eluates the degree of test reaction was stronger in the C-R system. However, titration endpoints of 6 quantified antibodies did not differ significantly. In 2 patients with slightly positive DAT, antibody typing was negative or not clear in the serum. In the corresponding eluates, an anti-K and an anti-JKa could be identified only by the C-R technique. In such instances we recommend to use the C R technique to prevent transfusion complications. PMID- 9417340 TI - [Typing of thrombocytic antigens on the DNA level and their value in diagnosis of alloimmunothrombocytopenias]. PMID- 9417341 TI - [Rh-D genotyping for exon 2, 5 and 7 of German and Japanese blood donors with sequence specific polymerase chain reaction]. AB - RHD genotyping from fetal cells was applied for the detection of the RHD gene in the fetus of immunized Rh-D-negative women. Additionally, RHD genotyping was applied for the characterization of Rh-D variants. Although 44 nucleotide substitutions are known to code for 35 amino acid differences between the RHCE and the RHD gene, only a few polymorphisms have been investigated yet. We investigated 7 RHD-specific nucleotides on exons 2, 5, and 7 with sequence specific primers and 1 nucleotide with ligation-based typing. All RHD genotyping results were correlated with serological results and established genotyping methods in 116 German and 98 Japanese blood donors, because different genetic sequences coding for Rh-D polypeptides have been described in different ethnic groups. Sequence-specific amplification of D-specific sequences was concordant with the serological result in all blood donors tested. However, ligation-based typing on exon 5 gave false-negative results in 7 donors. In summary, 5 new sequence-specific PCRs have been evaluated for further characterization of Rh-D variants. Furthermore, the methods described allow nested PCR and thus may help in determination of the fetal RhD status from maternal peripheral blood during pregnancy. PMID- 9417342 TI - [Routine PCR screening for HBV, HCV and HIV-I genome in a large blood donation services--experiences and initial results]. AB - We adapted the PCR method to screen up to 3,000 blood donations per day for HBV, HCV, and HIV-1 contamination. Concerning logistics: The first step is the generation of 3 identical microtiter plates (PT) by using the self-validated automatic sample processor with disposable tips. Using the first PT, we pooled up to 600 aliquots taken from blood donations which are serological negative and free for clinical usage according to actual federal regulations. In the case of a positive PCR pool result the viremic donation is identified by 2 additional PCR pool testing steps with smaller pool sizes using the second and third PT. All described steps are supported by electronic data processing. The PCR-method: After virus concentration by ultracentrifugation and--in case of HCV and HIV-1- additional reverse transcription PCR-amplifications were performed. PCR in two genomic regions are done for each virus. Laser-induced fluorescence detection after polyacrylamide gel electrophoresis and computer analysis were used to check the amplification products. Using this approach, a virus-containing donation can be detected in up to 599 negative samples with following sensitivity: HBV and HIV 1, 1,000-1,500 genome equivalents/ml; HCV, 2,000-2,500 genome equivalents/ml, thus sensitivity being in the range of commercial available PCR kits when testing nonpooled samples. The sensitivity was validated by using national and international available standards with known virus genome concentrations. All processing steps are checked using different controls such as, e.g., negative, positive, premix controls, reporter virus, inhibition controls. Routinely employing this validated methodology, we investigated 327,013 donations until the end of June 1996. During this survey, we found at least 16 virus-containing donations which are negative in corresponding serological tests and would have been transfused (4 HBV-, 13 HCV-, 0 HIV-1-containing donations), including one later seroconversion for HBV and one for HCV. Using our adapted PCR-methodology, it seems possible to shorten the diagnostic window periods with acceptable costs for large transfusion centers (15 DM per donation for all 3 viruses including all steps and investments). Therefore, PCR seems to become a new method of choice to prevent transfusion transmitted infections. PMID- 9417343 TI - [D-category VII depends on amino acid substitution Leu(110)Pro]. AB - A point mutation has been postulated as cause of the phenotype D category VII, based on data of 3 probands only. Repeatedly, D protein variants have been found to be due to heterogenous molecular events. Therefore, the aforementioned cause was to be tested with more probands. In a systematic study, 68 nonrelated probands with D category VII were found. 33 were selected by chance, and the nucleic acid region 280-329 was sequenced after PCR amplification. All examined probands showed the postulated Leu(110)Pro substitution. No further polymorphisms were detected. Our data show that in Southern Germany D category VII is homogenously due to the amino acid substitution Leu(110)Pro. This allows the exploitation of this polymorphism for the prenatal detection of D category VII and the related Tar antigen. PMID- 9417344 TI - [Specific Lp(a) apheresis for secondary prevention of arteriosclerosis]. AB - Lipoprotein(a) (Lp(a)) is an independent risk factor for arteriosclerosis. It consists of the Lp(a)-specific apo(a) which is bound to the apo-B of an LDL particle by a disulfide bridge. Apo(a) is homologous to parts of the plasminogen molecule: It consists of one kringle 5 and 10-40 kringles 4 of the plasminogen molecule. Due to the lack of alternative drug treatment, 3 patients with early onset of arteriosclerosis, rapid progression, and elevated Lp(a) as their dominating risk factor were treated weekly with specific Lp(a)-aphereses. Since October 1992, we carried out 229 immunoadsorptions (IA) with specific columns containing anti-Lp(a) antibodies covalently bound to sepharose. To reduce Lp(a) from preapheresis values of 142 +/- 53 mg/dl to 25 +/- 11 mg/dl immediately after apheresis, we had to adsorb 1.4-3 patient's plasma volumes. Lp(a) rise to preapheresis values took 3-4 days. Protein reduction caused by loss of plasma during column changes remained tolerable (total protein before IA: 71 +/- 4 g/l, after IA: 56 +/- 4 g/l. Immediately after IA, these values were measured after the application of 991 +/- 207 ml of ACDB with 5,000 IU of heparin as anticoagulant. Hemoglobin remained unchanged (before IA: 13.4 +/- 1.4 g/dl, after IA: 13.6 +/- 1.5 g/dl). Side effects were mainly flush and tachycardia. They were seen especially when using new columns and plasma flow rates above 55 ml/min and were immediately reverted by interrupting the IA. Control angiography performed after 2 years in 2 patients showed no progression of disease, in the 3rd patient, the stress test showed a significant improvement as did clinical parameters. In our hands, IAs are a safe and efficient method for Lp(a) reduction and secondary prevention of myocardial infarction. Therapeutic efficiency should further be proven by a controlled trial. PMID- 9417345 TI - [Clinical and flow cytometry outcome of improved photopheresis methods in dermatologic patients]. AB - In photopheresis, 8-methoxypsoralen (8-MOP) is added to a mononuclear cell concentrate and then activated by UVA light, thus forming covalent bonds between DNA strands. Infusion of these modified cells that are not able to replicate any more seems to lead to the elimination of pathogenic T-cell clones and clinical improvement in patients suffering from cutaneous T-cell lymphoma, graft versus host reactions, and various autoimmune diseases. The original method described by Edelson et al. (1987) was improved by the following modifications proposed by Andreu et al. (1994): i) by using a cell separator (COBE Spectra) that produced purer concentrate with less red blood cells absorbing UVA light. ii) By applying 8-MOP directly into the collection bag, the drug side effects due to the oral application of the thousandfold dose and varying serum levels were avoided. iii) By irradiating the concentrate after collection, all cells received the same irradiation dosage. We treated 2 men with cutaneous T-cell lymphoma, 2 women with atopic eczema and 1 man with severe pustular psoriasis with overall 123 extracorporeal photochemotherapy (ECPC) sessions. In addition to routine laboratory analysis, detailed characterization of lymphocyte subpopulations was carried out by flow cytometry to differentiate T-helper and T-suppressor cells and their activation (HLA-DR, CD 25), B, NK cells, and monocyte subpopulations. The following mediators soluble were analyzed as well: Il-2-receptor and as indicators of acute phase reaction Il-8, neopterin and Il-1 beta. We observed a good clinical improvement independent of no significant trend in the immunological phenotype of circulating blood leukocytes. Our results suggest that ECPC effects are not mediated by a systemic immune response or alternatively are not measured in the blood compartment. PMID- 9417346 TI - [HBV-DNA positive findings in HBsAg negative blood donors and patients]. AB - There have been repeated discussions as to whether the implementation of anti-HBc screening of blood donations in Germany would be useful. We present several cases of HBsAg-negative patients and blood donors in whose plasma HBV-DNA was mainly found only after enrichment of virus particles by ultracentrifugation. In the case of a seroconverting blood donor, 3 of 4 HBsAg tests could not detect HBsAg. 90% of our HBsAg-negative, but HBV-DNA containing samples (n = 10) were positive for anti-HBc. The 'nested' PCR without previous ultracentrifugation was positive in only under 30% of the samples. Ultracentrifugation is an expensive method and will not be practicable for the testing of all blood donations. We conclude that the HBsAg tests which are available should be subjected to improvement. In addition, the implementation of anti-HBc screening could decrease the risk of posttransfusion HBV infection more effectively than PCR testing from nonenriched serum. PMID- 9417347 TI - [Therapeutic thrombocytapheresis--effect on hemorheologic parameters]. AB - Common features of all myeloproliferative diseases (CMPE) are a markedly increased number of platelets and restrictions in the subjective feeling of the patients, comprising symptoms like nausea, headache, sensory deficits and transient paresis. Meanwhile, it has been shown that platelet pheresis (mTD) does not only reduce platelet counts sufficiently but also results in an impressive relief of the subjective complaints. To test the part that rheological mechanisms play hereby, relevant rheological parameters in the blood of 22 CMPE patients and of 8 healthy platelet donors as controls were analyzed before and after treatment with an AS-104 cell separator. Concerning the viscosity of whole blood und the filterability of erythrocytes, there was seen neither a difference between the patients and the controls, nor before and after mTD. As a result of treatment of patients, the moderately raised plasma viscosity was normalized, whereas the aggregation of erythrocytes was significantly lowered. It is concluded that there is an influence of hemorheological conditions on the patient's subjective feeling. These would originate in neurological dysfunctions caused by CMPE induced restrictions in cortical microcirculation. The erythrocyte aggregation, lowered by mTD, would cause an improvement of microcirculatory fluidity and would, in consequence, abolish the neurological symptoms. PMID- 9417348 TI - [Heparin-induced thrombocytopenia type II: a clinically relevant problem?]. PMID- 9417349 TI - [Intrauterine transfusion in fetal alloimmunothrombocytopenia: comparison of maternal and fetal weight-adjusted IgG therapy with exclusive fetal thrombocyte transfusion]. AB - Fetal alloimmune thrombocytopenia is caused by maternal immunization against a fetal platelet antigen and transplacental transfer of the antibody into the fetal circulation. Since 10-20% of the fetuses or newborns are threatened by intracranial hemorrhages, early management is required. Fetal blood sampling should be started between the 20th and 22nd week of gestation to assess fetal phenotype and platelet count. Different concepts to elevate the fetal platelet count have been discussed: maternal intravenous immunoglobulins, fetal intravenous immunoglobulins, or only repeated fetal platelet transfusions. Our investigations suggested that platelet transfusions in short intervals appear to be the only effective regimen to increase platelet counts in thrombocytopenic fetuses at risk. PMID- 9417351 TI - [Inline filtration of erythrocyte concentrates with the Leucoflex LCR4 T/B system]. AB - The in-line-filtration of red cell concentrates (RCC) is an effective method to reduce white-cell-induced reactions. SAG-M RCCs were prepared in quadruple top/bottom bag systems and filtered after storage at 4 degrees C (study: n = 12). The mean white blood cell (WBC) reduction was 99.9% with the number of residual WBCs at about 0.67 x 10(6) per unit and with a loss of Hb mass of 10.64%. No cell fragments of WBCs and monocytes were detected (MAIPA). After storage of 42 days, mean values of hemolysis level (0.64%) and ATP concentration (2.27 mumol/g Hb) were improved compared with unfiltered RCCs. Comparably good results of leukocyte depletion were achieved by preparation of about 3,000 in-line filtrated RCCs for clinical use, measured by routine quality control (n = 25). Our experience shows that the systems are easy to handle and to use. The Leucoflex LCR4 in-line T/B system (Maco Pharma) can be recommended in the clinical practice by virtue of the effective leukocyte elimination, particularly in those blood banks with platelet concentrate production from buffy coats beside the RCC in-line filtration. PMID- 9417350 TI - [Plasma donor screening and product safety]. AB - In addition to the specific virus reduction and inactivation procedures during the manufacturing process, the thorough selection of the source material is an essential factor for the product safety of plasmaderivatives. Screening of donors, inventory hold, and plasma pool testing by PCR are important aspects in raising the safety margin. Between January 1, 1994 and June 30, 1996, 576,673 plasma donations were collected in the German and Austrian plasmapheresis centers of IMMUNO. Incidence rates for viral markers (confirmatory tests) as antibodies against HIV, HCV, and HB surface antigen (HBsAg) were calculated as 0.35, 0.87, and 0.87 per 10(5) donations, respectively. Due to look-back, 1.33% of the donations were eliminated between January and October 1995. In 32% of the donations being rejected in the time of inventory hold in case of elevated alaninamino-transferase (ALT) of the donors, HCV genomes were shown which emphasizes the importance of this surrogate marker. Out of 1,240 pilot pools tested by quality assured PCR (IQ-PCR) being performed by IMMUNO since October 1995, in 4% of cases HCV and in 0.2% HBV genomic equivalents were shown. However, genomes of HIV were not found. The measures aiming at a safe plasma pool are part of IMMUNO's safety concept which is confirmed by the fact that there was no transmission of hepatitis or AIDS viruses by IMMUNO products, since virus inactivation procedures have been established. PMID- 9417352 TI - [Anti-hepatitis C prevalence and incidence in 2.8 million blood donors in Lower Saxony--residual transfusion-associated HCV risk]. AB - 561 out of 2,777,021 blood donations from 582,655 donors tested confirmed positive for anti-HCV (RIBA II or Matrix Dot), 549 of them at their first donation, in their first HCV test ever, or in the first HCV test with a new, more sensitive test generation. Thus, our anti-HCV prevalence is 0.037% or 1 in 2,679 donations; among first-time donations it is five times higher than among repeat donations. Twelve repeat donors seroconverted, yielding an anti-HCV incidence of 0.0008% or 1 in 122,570 repeat donations and a seroconversion rate of 2.32 per 10(5) repeat donor person-years. The residual risk associated with transfusion of blood for repeat donors amounts to 5.2 per 10(6) repeat donations. We estimate a risk reduction from introduction of direct virus genome testing after PCR to be at 3.7 per 10(6) repeat donations. Look-backs among recipients of the last seronegative blood products from 12 donors with subsequent seroconversion revealed no recipient infection. Thus, today the residual risk for HCV transmission through transfusion of blood components obtained from Lower Saxony blood donors is quite low. The added safety from direct virus genome testing after PCR is considered extremely low, which casts doubt on the cost effectiveness of such a measure. PMID- 9417353 TI - [Effect of whole blood preparation and leukocyte filtration on storage of erythrocyte concentrates over 42 days]. AB - Leukocyte reduction prior to storage of red cell concentrates (RCC) may reduce the incidence of HLA alloimmunization and may improve the quality of stored RCC. We tested an RCC leukoreduction filter system (Baxter) with an integrated Pall RCM-1 filter and investigated the filtration efficiency and the impact on red cells during storage for 42 days after different whole-blood preparation procedures. After whole-blood donation, all units (n = 9, +6 unfiltered controls per group) were either stored at 22 degrees C for up to 6 h (groups 1, 2, 3) or for 24 h (group 4) RCC were either prepared from a triple blood bag system (PL 146, groups 1, 2) or were buffy-coat-depleted (PL 2209, groups 3, 4). Groups 1, 3 und 4 were filtered immediately, whereas group 2 was stored another 18 h at 4 degrees C before filtration. Filtration efficiency and filtration time were determined. Hemolysis, ATP, 2,3-diphosphoglycerate (2,3-DPG), glucose, electrolytes, lactate, hematocrit, Hb and pH were quantified weekly. White blood cells (WBC) were reduced by 3-4 log10 to 0.1-0.3 x 10(6) (mean) by filtration (all groups) regardless of the RCC preparation. Mean filtration times were 1 h 36 min, 33 min, 29 min, and 18 min for the groups 1 to 4, respectively. There were no major differences for the in vitro storage values except for hemolysis and pH, which were elevated in all filtered units, and potassium, which was elevated in the unifiltered units. In conclusion, prestorage leukocyte filtration of RCC reduced the WBC by 3-4 log10, whereas the extended filtration time was a major disadvantage. PMID- 9417354 TI - [Electron microscopy study of HES cryopreserved erythrocytes]. AB - Using electron microscopy (EM), it was examined whether cryopreservation with HES causes shape changes of erythrocytes. Each of 11 erythrocyte suspensions (Hct = 40; HES 200,00/0.5/12.5%; 60 mM NaCl) was separated into 40-ml samples, cooled down to -196 degrees C and finally stored. In addition, 11 samples were stored at -80 degrees C for 3 months. The preparation for EM was done immediately after thawing or in case of native cells shortly after donation. On EM micrographs, there was no visible difference between native and cryopreserved erythrocytes. In every case the preparations showed normocytes, either as single cells or having attracted other ones, forming rouleau. Packed cells were attached tightly to each other without any gap in between. The tangent count method neither revealed an excess of convexity nor of concavity. The erythrocyte membrane looked normal, and the cytoplasmatic space was filled with electron-dense material (hemoglobin) homogeneously; Heinz bodies were not seen. Scanning microscopy portrayed native as well as cryopreserved cells as discocytes with the characteristic bioconcave resting shape of human erythrocytes. It is concluded that cryopreservation of erythrocytes with HES does not cause shape changes. Therefore, a sequestration into the reticulo-endothelial system (RES) by means of identification of morphological abnormalities may not be expected. PMID- 9417355 TI - [Cryopreservation of thrombocyte concentrates: results with a new anticoagulant]. PMID- 9417357 TI - Proceedings of the International Congress on Hepatocytes. Applications in cell biology, toxicology and medicine. Tubingen, Germany, September 25-28, 1996. PMID- 9417356 TI - [Quality control in producing thrombocytapheresis preparations with minimal leukocyte contamination]. AB - Passenger white blood cells (WBC) in platelet concentrates (PC) produced by apheresis can cause a variety of adverse effects in recipients after platelet transfusion. With new technologies (LRS, leukocyte reduction system), the preparation of PC with a low WBC contamination is possible without consecutive filtration. In a prospective examination, we compared the effect of LRS apheresis on the donor, the quality of the resulting PC (n = 120), and the platelet increment in the corresponding recipients with conventionally prepared PC (n = 27). In LSR apheresis, no serious adverse effects on the donors were observed, but the post-donation absolute numbers of lymphocytes were reduced from 1,787 +/- 505/microliter to 1,402 +/- 383/microliter (p < 0.001). Comparable results were observed in non-LRS donors. The collection efficiency of the LRS procedures was 50.0 +/- 7.6%, resulting in a yield of 4.25 +/- 1.03 x 10(11) platelets/PC. Flow cytometric examination concerning the expression of platelet glycoproteins in LRS PC showed no elevation in mean fluorescence of CD 62 (6 +/- 4) or CD 63 (9 +/- 3) in comparison to non-LRS PC (mean fluorescence CD 62: 7 +/- 4, CD 63: 8 +/- 3). Median leukocyte contamination of the LRS PC was 0.41 x 10(5) (range 0.06-8.5 x 10(5)) WBC/unit. In 43 recipients, the 24-hour corrected count increments (CCI) after transfusion of LRS PC (12,530 +/- 8,761) showed no significant differences when compared with the CCI in 20 recipients of non-LRS PC (13,133 +/- 9,812, p = 0.75). LRS apheresis seems to be safe procedure, resulting in PC with low WBC contamination, which causes adequate CCI values after transfusion. PMID- 9417358 TI - [Surgery in Switzerland]. PMID- 9417359 TI - [The surgeon as modern demi-god]. PMID- 9417360 TI - [Surgery--a res publica]. PMID- 9417361 TI - [The surgeon and the media]. PMID- 9417362 TI - [The surgeon in social communication]. PMID- 9417363 TI - [Public image of the surgeon as a legal problem]. PMID- 9417364 TI - [Surgery and public relations--guidance in communicating in different ways]. PMID- 9417365 TI - Shared mental health care in Canada. PMID- 9417366 TI - Impact of patch testing on dermatology-specific quality of life in patients with allergic contact dermatitis. AB - BACKGROUND: A dermatology-specific quality-of-life instrument has been created and validated for its sensitivity, reproducibility, content, and construct validity. This instrument was used in an observational prospective study that evaluated the cost-effectiveness of patch testing in patients with contact dermatitis and suspicion of allergic component. OBJECTIVE: This multicenter, prospective, observational study was designed to study the various direct and indirect costs associated with diagnosis and treatment of suspected allergic contact dermatitis and the benefits of diagnosis with and without patch testing. The costs and benefits of various methods of diagnosis are evaluated to determine if patch testing is more cost-effective as a diagnostic method. One of the outcomes evaluated was the quality of life of the participants in the study and the impact of patch testing on this cohort. METHODS: A total of 567 subjects were enrolled to obtain evaluable data from at least 500 subjects from ten study centers; the investigators were chosen so as to obtain a mix of stratified degrees of usage of patch testing. Data were collected on demographics, physical characteristics, history of disease, visits to physicians, and, to evaluate costs, the use of resources such as drugs, nondrug substances, and services; other outcome data collected were on physician evaluation of improvement, confirmation of diagnosis, physician opinion of the disease management, and patient evaluation of dermatology-specific quality of life (DSQL), and pertinent economic factors. Only 6-month and 12-month follow-up visits were mandatorily scheduled according to the protocol; however, participants and providers were free to arrange additional visits as deemed necessary for health care. RESULTS: This report focuses on the evaluations at the 6-month follow-up. There are completely evaluable data on pharmacoeconomics and DSQL for 431 patients. About 43% were patch tested, and the rest were diagnosed using the information from history and physical examinations. There was significantly better improvement in each of the DSQL domains in patch tested subjects compared with non-patch-tested subjects. In addition, patients treated in "low" use clinics had lower quality of life at 6-month follow-up than those treated in "high" use clinics. CONCLUSION: Patch testing helps to diagnose the etiology of contact dermatitis early and treat the disease before it becomes chronic, thus reducing resources used and improving patient quality of life considerably. PMID- 9417368 TI - NIH Technology Assessment Workshop on Alternative Medicine: Acupuncture. Gaithersburg, Maryland, USA, April 21-22, 1994. PMID- 9417367 TI - A "new" cosmetic allergen. PMID- 9417369 TI - [Priorities in public health: from one side of the Atlantic to another]. PMID- 9417370 TI - [Plague in India in 1994: is there a world threat from this focus?]. AB - The plague epidemic in India in 1994 has set off an international alarm signal. After describing plague through an epidemic view, this study tries to deal with the characteristics of this indian epidemic and the measures taken as local and international levels. This work also questions us concerning the real risk of a plague epidemic from the indian source. PMID- 9417371 TI - [Plague epidemic in India from September to October 1994: what lessons can be learned from a study of press coverage?]. AB - During September 1994, the french press and the foreign one have published abundant news about the plague which broke out in India and also because of the threat of an epidemic towards the others EC countries. From September 24th to October 9th, 1994, the author has collected the whole available daily papers in particular all editions of the eight national papers and foreign daily ones stating the information hitting the headlines. The review of these papers has shown an intensive media coverage in the french press overall from September 28th to October 1st, 1994, accompanied by an evolution of concerned items which is first of all a completely indian problem whose scale is finally threatening over the world. Due to this situation, some countries have taken measures of extreme protection. PMID- 9417372 TI - [General practitioners and doping in sports: knowledge and attitudes]. AB - Most campaigns of prevention from doping (use of certain substances by athlete that could have an effect like an artificial improvement of his (or her) physical and/or mental conditions) have been relied on the physicians considered as main actors in this field. However do the physicians have the necessary knowledge and attitudes and do they simply wish to take part pertinently in actions of doping prevention? This study has been conducted in order to examine their knowledge and attitudes in front of doping, to evaluate their role in the prevention campaign. So 280 french physicians have randomly been selected and interviewed by telephone. The response rate was about 62%. The selected population was made up of 173 physicians in which were 122 males and 51 females, the average age was 44.5 +/- 6.5 years. The questioned physicians seemed to have a few knowledge about doping, in particular through the eight families of prohibited substances in which they only mentioned anabolic steroids and amphetamines. Only 50% of them think doping can also concern children, therefore 33.6% had have to face this problem in the latest 12 months. For 86.5% of the questioned physicians, doping is a serious public health problem and 60% of them consider it as a kind of drug addiction. According to 60.9% of them, campaigns of prevention from doping are inefficient; and 92.5% would like to take part in it but 83.3% of them do not feel themselves prepared enough. The results are always similar with or without sports medicine diploma. Three different families of the physicians' attitudes are described. The physicians' involvement in doping prevention claims an action in two parts: to change their behaviours facing to doping they have to consider as any health issue, to provide them with prevention efficient "tools" for their sports patients. At last, considering doping as a public health issue and not only as a marginal practice restricted to the athletic elite is really essential. PMID- 9417373 TI - [Evaluation of risks of glycol ethers for the reproductive health]. AB - Glycol ethers (Ge) are a family of substances with a growing use in industrial and domestic products for the two last decades. Ge (group 1 and 2) are experimentally toxic for reproduction and development, at various levels. That begins to be found in humans. Epidemiological studies confirm toxicokinetic data showing humans are more sensitive than animals, because of a low excretion rate of the toxic metabolites. Occupational and consumer exposures are frequently higher than reference concentrations deducted from animal data. They may be involved in the growing number of genital reproductive system and reproduction anomaly, observed in most developed countries. Genetic toxicity is suspected from experimental data, but further investigations are needed. PMID- 9417374 TI - [Evaluation of the use of tobacco restrictions in French enterprises]. AB - This survey evaluates the respect of tobacco legislation at workplace among a random sample. It shows that a small and limited information can increase the awareness of the companies' managers. More than one out of three companies replied that new measures were implemented. In the majority of cases (more than three out of four) members of the board of directors believe that all the measures are well respected. A little bit less members of the employees are of the same opinion, with an exception for the smoker's lounge. Many of them believe, however, that the smoking restrictions are respected. PMID- 9417375 TI - [Knowledge and beliefs of 4th-year medical students concerning professional practices in general medicine. Results of a survey conducted at the School of Medicine of Brest]. AB - As part of preparations for the first seminar in general practice, organized for medical students in their forth year at the Brest faculty of medicine, students were asked to fill in a questionnaire concerning their beliefs and knowledge concerning general-practice medicine. Fifty-two students, or 89.6% of the class, completed the questionnaires. The results showed that despite the lack of formal contacts with this type of practice during their studies, the students had a good idea of what general-practice medicine is and of the role of the general practitioner in our health-care system. However, this study has illustrated the difficulties students experience in defining their future professional direction, since they receive no information during their training on the different options in medical practice. PMID- 9417376 TI - ["It's free, therefore they abuse...". Study of answers of medical students concerning drug consumption]. AB - In 1994, during a public health examination, in Nancy, a question was asked to students in their 6th year of medical studies: "why do the patients who benefit of a total reimbursement of the drug's cost (that is patients that receive free medical care because they have a serious illness), consume obviously more than the others?". The aim of this study is to analyse the student's answers: 57 students gave the correct answer (serious illness explains high consumption), 44 gave several answers (included free care explains high consumption); for 71 students, it is only an abusive consumption. Although the time to complete the test is limited, it is a good indication of the student's attitudes. Many of them do not make a difference between high consumption in a case of serious illness and abusive consumption as a result of free medical care. Then the student's attitude towards the patients is frequently negative. PMID- 9417377 TI - [Public health education integrated in hospital. An internship proposal, "Medical information and pharmacology"]. AB - According to a recent circular reforming french medical studies, we propose a teaching of medical information and pharmacology in situ within hospital instructions. Students could acquire an investigation methodology on the medicine economy. It will cover in four sessions the succeeding stages of medical information processing and be subject to an assessment: case studies and appreciation on student's, instruction record. By combining public health teaching with clinical practice, our project promotes its development in contact with other learnings and activities such as clinical research. PMID- 9417378 TI - [Evaluation of a tool for developmental disorder screening in 4-year-old children]. AB - With the aim of helping physicians working in a "Mother and Infant Protection Service" (PMI) in their mission of children's development difficulties screening, authors tried to assess performances of a clinical tool including parental informations and standardized tests. The whole four years old children going at school in a canton of Calvados department were independently examined by a PMI doctor using the tool to be estimated and by a specialized service working from a beforehand established protocol. For the language disorders on the one hand and the adaptation on the other hand, this evaluation shows a significant correlation between the PMI screening and the specialists one's: the sensitivity of the PMI examination is about 70% for those two types of difficulties with a positive predictive value respectively 28% and 57%. However the tool appears less reliable for the psychomotor disorders screening: only one quarter of the children who require a specialized investigation about these problems have been detected by the PMI screening. A filing and a selection of the questionnaire items should permit to improve the efficiency of this tool. PMID- 9417379 TI - [Optimal tumor therapy with cytoprotection by ETHYOL (Amifostine). 32nd Annual Session of the American Society of Clinical Oncology in Philadelphia 18-21 May 1996]. PMID- 9417380 TI - [Differential diagnosis of acute testicular pain using color-coded duplex ultrasonography: difference between testicular torsion and epididymitis]. AB - OBJECTIVE: To assess retrospectively the accuracy of colour-coded duplex sonography (CCDS) in distinguishing testicular torsion from epididymitis as the cause of acute testicular pain. PATIENTS AND METHODS: The results of CCDS were analysed for all 81 patients (mean age 27.2 years [6 weeks to 60 years]), admitted between 1.1.1995 and 30.6.1996 with the diagnosis of acute testicular pain. Testicular torsion was diagnosed when CCDS failed to detect perfusion in one testis. Regular arterial and venous perfusion of both testes excluded torsion. Epididymitis was diagnosed when hyperperfusion of the epididymis was demonstrated by CCDS. RESULTS: 20 of 22 cases of torsion, subsequently diagnosed at surgery, had been correctly diagnosed by CCDS (sensitivity 90.9%, specificity 98.3%). 55 patients had epididymitis, confirmed by the clinical course and follow up having excluded torsion. Other causes (trauma, tumour, inguinal hernia) were found in the remainder of patients. CONCLUSION: With a positive predictive value of 95.2% and a negative one of 96.6% CCDS is a highly suitable method for recognizing or excluding testicular torsion and thus clarifying the cause of acute testicular pain. PMID- 9417381 TI - [Perforation of the esophagus after esophageal manometry]. AB - HISTORY AND FINDINGS: A 75-year-old man was admitted for oesophageal manometry because of dysphagia for the past 2 years and retrosternal burning sensation unrelated to exercise. His general condition was appropriate for his age. INVESTIGATIONS: An oesophagogram showed corkscrew-like deformation of a diffuse oesophageal spasm. The first, but incomplete, manometry recorded clearly propulsive contractions with markedly raised and prolonged pressure, as in "nutcracker oesophagus". The lower oesophageal sphincter could not be demonstrated initially. Subsequent pH measurements provided no evidence for increased gastrooesophageal reflux. TREATMENT AND FURTHER COURSE: After the first manometry conservative treatment was initiated with molsidomine, nifedipine and nitrospray sublingual, but the dysphagia was not significantly improved. A second manometry was performed before a planned surgical exploration. Placing of the catheter was again difficult and mild resistance experienced. Endoscopy revealed only minimal, presumably superficial, mucosal lesions. 2 days later bilateral pleural effusions together with mediastinitis occurred. Conservative treatment was continued until finally a distal oesophageal perforation was demonstrated. At surgery the perforation was seen and a oesophagectomy with gastric pull-through and intrathoracic anastomosis performed. However, the patient died of septic multi-organ failure. CONCLUSIONS: Oesophageal manometry is a safe but invasive method with few complications for measuring oesophageal motility. Although this has not previously been reported, oesophageal perforation with mediastinitis may end fatally, if the particular circumstances are unfavourable. In addition to special anatomical features, type and state of the manometric catheter may present a risk factor. PMID- 9417382 TI - [Chronic right heart failure after implantation of a cava filter]. AB - HISTORY AND CLINICAL FINDINGS: A 39-year-old woman complained of dyspnoea and increasing abdominal pressure sensation. A Greenfield filter had been implanted into her inferior vena cava (IVC) 4 years previously because of pulmonary embolism from a deep vein thrombosis after a hysterectomy with abscess formation. Physical examination revealed neck vein congestion, jaundiced sclerae, a tense abdominal wall, ascites and a soft machinery murmur in the paraumbilical region. INVESTIGATIONS: Transaminase activities were slightly raised (GOT 38 U/I, GPT 20 U/I), but total bilirubin and direct bilirubin were markedly elevated (2.9 mg/dl and 1.1 mg/dl, respectively). There was no evidence of cholestasis or decreased liver synthesis. Ultrasound showed marked dilatation of the IVC and hepatic veins, and echocardiography revealed right ventricular enlargement with grade II tricuspid regurgitation. Calculated pulmonary arterial systolic pressure averaged 50 mmHG. Colour-coded Doppler sonography demonstrated an aorto-caval shunt at the level of the filter in the IVC and penetration of a filter strut into the aortic lumen. TREATMENT AND COURSE: After removing the ascitic fluid by fluid and sodium restriction, and administration of an aldosterone antagonist and a loop diuretic, the A-V fistula was closed surgically and the filter removed. Three months after operation she was put on phenprocoumon (Quick value 20-30%). At the latest outpatient examination, 6 months after the operation, she was free of symptoms. CONCLUSION: As filter implantation in the IVC may produce severe complications, indications for it need to be demonstrated by further studies of its efficacy. PMID- 9417383 TI - [Performance and postoperative care of heart transplantation]. PMID- 9417384 TI - [Cytokine determination. Diagnostic significance from the clinical and immunological viewpoint]. PMID- 9417385 TI - [Thomas Mann (1875-1955) and pneumology. Indication for thoracic surgical intervention in April, 1946]. PMID- 9417386 TI - [Effectiveness of basis resuscitation]. PMID- 9417387 TI - [Renal cell carcinoma]. PMID- 9417388 TI - [Stress, anxiety and and event related potentials]. AB - Our aim was to study the relationships between stress and anxiety with both psychological and neurophysiological (Events Related Potentials: CNV and P300) methods. The study was divided into 2 parts. In the first part, the research was carried out among 32 out-patients suffering from anxiety disorders (generalized anxiety disorders) according to DSM IV. All of them were drug free and displayed scores higher than 45 on the Spielberger Anxiety State and Trait Scale. They were compared to 40 controls paired in age and sex. The 2 groups displayed a score higher than 200 on the Amiel-Lebigre Life Events scale. In the second part, the control subjects were divided into 2 sub-groups The first one displayed scores higher than 45 (anxious controls) on the Spielberger Anxiety-State Scale while the second one displayed scores lower than 45 (non anxious controls). Two ERPs were recorded, the P300 by using the classical "Oddball" experimental paradigm in auditive modality and the Contingent Negative Variation (CNV) by using a reaction time task with warning stimulus. The results showed not only clear differences between the subjects who suffered from anxiety and the controls but also showed opposite results between anxious out-patients (anxiety disorders) and anxious controls. The non anxious controls were intermediate. While the outpatients showed a decreased P300, the group of anxious control showed an increase of this potential. The first one displayed a CNV/M1 (contingent negativity variation/early part) increase and a longest reaction time, while the second one exhibit an early CNV decrease and normally reaction time. It appears that the stress response expressed itself differently according to the psychological state and the stress situation. The behavioral and neurophysiological data will be discussed in the framework of cognitive, behavioral and psychophysiological theories. PMID- 9417389 TI - [Use of methylphenidate in adults with attention deficit disorder with hyperactivity]. AB - Attention deficit, hyperactivity disorder was recently described in adults. The clinical individualization of this syndrome progressed but the categorical approach remains to be entirely completed. The residual form of the childhood disorder does not generate diagnostic problem when childhood previous history is known. ADHD without childhood history refer us to retrospective difficulties of diagnosis and various evolutions of the infantile form linked or not, with other psychiatric pathologies. Etiology remains unknown, hypothesis of an hereditary disfunction of neurotransmitters is the more studied. These patients can benefit from a psychostimulant treatment. Four controlled studies with methylphenidate demonstrated to be significantly superior to placebo. Even if there are methodological difficulties not resolved (comorbidity, homogeneous population) results are encouraging. PMID- 9417391 TI - [Sleep during post-partum depression]. AB - BACKGROUND: Numerous studies have been published on postnatal depression (PND) over the last 10 years. A controversy has arisen regarding the specificity of the diagnostic concept. It is based on 2 points: the hormonal environment and the course and recurrence of PND. EEG Sleep studies are also numerous, but this paper presents the first study on EEG sleep profile during PND. METHODS: 24 women suffering from major depression according to RDC were placed in 3 groups: 1) Group A (n = 8): PND; 2) Group B (n = 8): depression with a past history of PND; 3) Group C (n = 8): depression without a past history of PND. Women were age matched and according to the Hamilton 24 item severity score. Group A patients were delivered within less that 6 months, groups B and C within a minimum of 3 years. None were pregnant or alcoholic, and none were physically ill. RESULTS: There was no difference between groups B and C. Group A was characterised by a significantly longer stage IV sleep. There was also a strong tendency to a shorter stage I sleep and a better quality of sleep (total sleep time and number of awakenings) during PND. DISCUSSION: Our study shows that, even if similar to major depression, specific polysomnographic alteration can be found during post partum depression. This finding is relevant to the hypothesis of PND's diagnostic specificity. The perfect similarity between groups B and C strengthens this evidence. Nevertheless, the significance of SWS alterations are difficult to explain and additional studies are required. CONCLUSION: The EEG Sleep profile during PND differs from major depression of the same severity. This appears to favour the specificity of the diagnostic concept. PMID- 9417390 TI - [Effect of a coordinated management program on the outcome of a cohort of ambulatory psychiatric patients]. AB - This study describes a one year follow-up study of 53 ambulatory patients treated with case management oriented care. These patients, characterized by a chronic evolution for an average of 10 years, presented after a one year period a significant improvement of different symptomatic parameters (positive and negative psychotic symptoms). Psychosocial adaptation was also improved for their dependency, their activity and the amount of their social contacts. These results underline the usefulness of case management, which permits an optimal use of available therapeutic facilities. Moreover, the setting of such a program allows a more specifical treatment of this category of patients. PMID- 9417392 TI - [Negative symptoms, depression, anxiety and alexithymia in DSM III-R schizophrenic patients]. AB - Coined by Sifneos in 1972, alexithymia refers to a relative narrowing in emotional functioning, an inability to find appropriate words to describe their emotions, and a poverty of fantasy life. Although initially described in the context of psychosomatic illness, alexithymic characteristics may be observed in patients with a wide range of medical and psychiatric disorders: Parkinson disease, depression, anxiety, substance abuse and eating disorders. Flattening of affect and poverty of speech, major negative symptoms, referred to chronic schizophrenia: there is a lack of outward display of emotions. Accordingly, some disturbances of alexithymia's scores would be expected in schizophrenic patients. The aims of this study were: first to establish some correlations between alexithymia and some symptoms of schizophrenia, and second to estimate the intensity of alexithymia in negative versus positive and undifferentiated schizophrenic patients. Twenty-nine patients, meeting DSM III-R criteria for schizophrenia have been studied. All of them treated by neuroleptics, were in a stable clinical status for at least one month. The patients were assessed by one trained psychiatrist (IN) using six rating scales: Beth Israel Questionnaire (BIQ) for alexithymia, Positive and Negative Syndrome Scale (PANSS), Depressive Retardation Rating Scale (DRRS), Montgomery and Asberg Depression Rating Scale (MADRS), revised Physical Anhedonia Scale (PAS), and finally, Extrapyramidal Symptom Rating Scale (ESRS). In the total sample, the mean score of BIQ was 4.79 +/- 1.68 (mean +/- SD). Significant correlations were found between alexithymia and blunted affect (r = 0.376; p < 0.05), poverty of speech (r = 0.471; p < 0.01), anxiety (r = 0.370; p < 0.05), total score of DRRS (r = 0.370; p < 0.05), and motor subscore of DRRS (r = 0.429; p < 0.05). The patients with negative symptoms of schizophrenia had significantly higher total scores in alexithymia (p < 0.05), blunted affect (p < 0.0001), poverty of speech (p < 0.0001), anxiety (p < 0.05), total score of DRRS (p = 0.01) and his motor subscore (p < 0.0001) as compared to positive and undifferentiated subtypes. In our study, alexithymia seems to be correlated with negative and depressive symptoms in negative forms of schizophrenia, regardless of medication status. PMID- 9417393 TI - [Rett syndrome and autism. Early comparative evaluation for signs of autism using family movies]. AB - Rett's syndrome progresses in 4 stages: the first signs of the disorder appear after a period of 6 to 7 months, during which development is considered to be normal. This asymptomatic period is apparently an essential criterion of the diagnosis, but some parents have reported some prodromes. In stage II of the disease (before 3 years), signs common with autism dominate the clinical picture and the diagnosis of the latter was often formulated. Our working hypothesis is that the pedopsychiatric analysis of home movies of young girls with Rett's syndrome, taken by the parents before the age of 2, may be able to show early clinical signs. The present study involved examining home movies of children subsequently diagnosed as having Rett's syndrome (n = 9) in comparison to those of autistic (n = 9) and normal (n = 9) children, using semiological evaluation tools (IBSE, BFE). The persons scoring were not advised of the diagnosis. The observations were thus situated before the disorders and/or at the time of their appearance. The study confirms the asymptomatic interval between birth and the first signs of the disease, it defines the mode of onset and shows the disturbance of certain functions such as intent and imitation, more pronounced in Rett's syndrome children between 12 and 18 months. At this age, it also enables Rett's and autistic children to be differentiated on the basis of the different involvement of the "cognition" function and unusual posture, more pronounced in these girls. It does not, however, differentiate Rett's from autism between 6 and 12 months and it is thus not surprising that at this stage the diagnosis of rett's syndrome or autism may be a source of confusion. PMID- 9417394 TI - [Validation of an animal model of anhedonia, a major symptom of depression]. AB - One of the two core symptoms of human depression is anhedonia, the loss of interest or pleasure in daily activities. Daily stressful life events are recognized as predisposing factors in the etiology of depression. Rats submitted to a regimen of chronic, mild, unpredictable stress exhibit behavioral deficits consistent with a loss of responsiveness to reward, such as decreased sucrose consumption, decreased ability to associate rewards with a distinctive environment, and decreased sensitivity to rewarding electrical brain stimulation. Normal behavior can be restored by chronic treatment with tricyclics, atypical antidepressants, monoamine oxydase inhibitors and electroshocks, but not by other psychotropic agents such as antipsychotics. In addition, chronically stressed animals exhibit REM sleep abnormalities resembling those observed in depressed patients and recognized as biological markers of depression. These data provide evidence supporting chronic stress-induced anhedonia in rats as an original animal model of human depression combining convergent elements of biological, etiological, symptomatological and therapeutic validity. This realistic simulation of depression may prove useful for a better understanding of pathophysiological mechanisms involved in depressive disorders. PMID- 9417395 TI - [Comparison of 6 different methods for lorazepam withdrawal. A controlled study, hydroxyzine versus placebo]. AB - PATIENTS AND METHODS: 154 outpatients with generalized anxiety (DSM III-R criteria), followed by general practitioners, gave an informed written consent to participate in this multicenter, randomized, placebo controlled study previously approved by a legal ethic committee (CCPPRB). The patients had to be long term consumers (at least 3 months) of 2 mg daily of lorazepam and were withdrawn using transiently an antihistaminic anxiolytic (hydroxyzine or placebo TAD) according to 6 different procedures defining 6 parallel groups: hydroxyzine 50 mg, abrupt or progressive withdrawal; hydroxyzine 25 mg, abrupt or progressive withdrawal; placebo, abrupt or progressive withdrawal. Following this 4 week-period of withdrawal, the patients were without any treatment for a post-study follow up 2 month-period. Clinical evaluations for anxiety (HARS, Zung), sleep (Spiegel), BZD withdrawal syndrome (Tyrer), adverse reactions and clinical global impression (CGI) were performed at D0, D7, D14, D28, D35 and D88. Investigators opinion and patients attitude towards BZD were collected at D88. STATISTICAL ANALYSIS: Analysis of variance for quantitative variables and chi square test for qualitative or ordinal variables. RESULTS: Whatever abrupt or progressive, with or without hydroxyzine support, using half or full dosage, lorazepam withdrawal proved to be feasible even after a long term BZD treatment (mean = 64 months +/- 60). GPs opinion (72% satisfied: D35; 78% satisfied: D88) is satisfying but patients attitude (at D88: 54% patients desired to be regiven a tranquilizer, 31% patients occasionally had a BZD and 22% formally demanded a prescription of BZD) is more questionable. Despite a high initial level of anxiety under lorazepam (HARS = 21 +/- 10 at D0), after a one-month period of withdrawal (under placebo or hydroxyzine) followed by a 2 month-period without any treatment, 75% patients were totally free of any drug and their level of anxiety was significantly decreased (D88: HARS = 12 +/- 9). Progressive withdrawal appeared preferable if compared to abrupt since the number of drop outs between D28 and D88 was less important and the procedure judged more favourably by the patients. Levels of anxiety significantly decreased in both the groups (progressive and abrupt) but sleep parameters and number of withdrawal symptoms between D7 and D28 were improved only in abrupt withdrawal group (p < 0.0001). Considering hydroxyzine, 2 patients dropped out between D0 and D28 in the group hydroxyzine 25 mg, 6 patients in the group 50 mg and 5 patients in the group placebo. Levels of anxiety (HARS et Zung) were significantly improved in hydroxyzine 50 mg group (p < 0.007) and in hydroxyzine 25 mg group (p < 0.012) but not in placebo group. Withdrawal symptoms (Tyrer) between D0 and D28 were improved only in hydroxyzine 50 mg group and the number of side effects was significantly improved in both the hydroxyzine (25 et 50 mg) groups but not in placebo group. However, no significant difference was found between the 3 groups. Daytime sleepiness is more frequent in hydroxyzine 50 mg group. DISCUSSION: These results proved a significant improvement of anxiety, a decrease of side effects in both the groups treated with hydroxyzine and a reduction of withdrawal symptomatology in hydroxyzine 50 mg group. When a patient is engaged to be withdrawn from of a lorazepam long term treatment, it can therefore be proposed as a support a transient prescription of hydroxyzine 25 mg TAD to markedly anxious patients and of hydroxyzine 50 mg TAD to patients presenting a withdrawal symptomatology. PMID- 9417396 TI - [Cognitive behavior therapy of agoraphobia, retrospective study of 345 cases]. AB - Epidemiological data of 533 (42% male) agoraphobic patients referred to an anxiety disorder unit are reported. Comorbidity with mood disorders was high (48%). Therapeutic response was studied in 345 patients who received cognitive behaviour therapy (CBT): 131 patients dropped out and 214 completed treatment. At post-test 71% of the completers were improved with maintenance of the gains in patients reevaluated at follow-up points ranging from 6 months to 18 years. Medication intake was significantly decreased. Intent to treat analysis of the whole CBT sample showed that 57% of the 345 patients were improved. Drop-outs were predicted by female sex and psychotic personality traits. This suggests that personality assessment should be carried out in agoraphobia and therapeutic interventions on personality considered in patients with disordered personality. The relatively low prevalence of women in the whole sample and their higher prevalence in drop-outs are discussed. PMID- 9417397 TI - [Use of electroconvulsive therapy in the adolescent]. AB - Despite the progress of pharmocotherapy, electroconvulsive therapy (ECT) is still used in a majority of countries to treat severe intractable mental disorders of the youth, yet few studies have been conducted to assess its use for individuals under 20-year-old. Efficacy, indications, side effects, technical characteristics and outcome are uncertain. A review of the 96 cases reported in the literature shows that: 1) its average frequency in adolescent psychiatric practice is similar throughout western nations and can be estimated around one ECT every year per million people; 2) intractable mood disorders, both manic and depressive episodes, are its main indications, since ECT treated more than 90% of the 66 cases reported; ECT can also offer an interesting alternative in some schizoaffective and schizophrenic episodes, in particular catatonic ones; 3) tolerance appears to be good, although secondary effects may occur. The most serious ones are infrequent spontaneous seizures and more common memory loss. Although no prospective studies are available on the evolution of cognitive side effects, they seem to disappear within a few weeks. PMID- 9417398 TI - [Quality of life of schizophrenic patients and clozapine]. AB - The authors review the various methods of evaluating quality of life in schizophrenics and note the limitations of that type of method in terms of both form and content. In practical terms, the patients' cognitive disorders may constitute a bias when the questionnaire is addressed. Using a 'subjective quality of life profile' by Gerin et al., the authors interviewed 22 patients presenting with 'recalcitrant schizophrenia' and treated with clozapine for more than 3 years. The patients were asked to assess the change in their quality of life by comparison with previous neuroleptic chemotherapies. For all the items investigated, the responses were in favor of clozapine. PMID- 9417399 TI - [Creativity and antipsychotic drugs]. AB - For the authors, the creative abilities of a schizophrenic patient are indicators of the therapeutic efficacy of an antipsychotic treatment and the incidence of adverse effects. The new antipsychotic drugs available, unlike conventional neuroleptics, show enhanced efficacy and do not exacerbate the negative symptoms. They may even alleviate them. With regard to clozapine, the authors illustrate the foregoing by two examples of a favorable course in two patients. Clozapine induced an indisputable clinical improvement and hence the opportunity for undeniable artistic activity. Lastly, the new antipsychotics contribute to changing the image that we have of schizophrenic patients. PMID- 9417400 TI - [Atypical pharmacologic characteristics of an antipsychotic drug: clozapine]. AB - The discovery of clozapine in the mid-sixties and the demonstration of its clinical efficacy vis-a-vis recalcitrant schizophrenia contributed to the development of specific psychopharmacological research addressing the concept of atypical antipsychotics. The research has a dual aim. First, identifying the action sites, in the brain, specific to clozapine and to conventional neuroleptics, in order to classify those drugs on the basis of the observed differences in pharmacoclinical profile (low incidence of neurological side effects, activity on schizophrenic deficiency symptoms, activity vis-a-vis certain forms of recalcitrant schizophrenia). Recent studies have used tools derived from molecular biology to determine the action sites. The results of those studies suggest that there are at least four classes of antipsychotics (reverse neuroleptics, conventional neuroleptics, atypical neuroleptics and atypical antipsychotics). The second aim of the research is to determine the behavioral effects of each of the recognized classes of medication in order to determine the neurobiological substrates specific to the elementary cognitive operations impaired in schizophrenia. There is preclinical evidence to suggest that, in each area investigated (low incidence of neurological side effects, negative symptoms, cognitive symptoms), clozapine, a "dirty" drug, acts on different neurotransmission systems. The research thus aims to determine the pharmacological profile of the drugs of the future, designed to treat the cognitive deficiencies specific to the various types of schizophrenia. PMID- 9417401 TI - [Social trajectory and schizophrenia]. AB - On the occasion of the Clozapine symposium we have had cause to reflect on the social life-histories of schizophrenic patients. After an analysis of a cohort of 40 patients aged over 65 years, whose medical records had been kept up since the start of their disease, we have set up a methodology which enables us to study the social life-histories of these patients. Our aim is ultimately to compare the course of the disease with the treatments received by these patients and the therapeutic structures they have been offered. For this purpose, we shall study three criteria of social exposure: 1) Autonomy, 2) Integration into the family, 3) Integration through work. The aim of the present paper is to record our initial analytical findings in these three years. PMID- 9417402 TI - [Medico-economic evaluation of treatment with clozapine versus treatment with previous neuroleptics]. AB - Since clozapine has been introduced into the treatment of schizophrenia, pharmaceutical costs rised in hospital. However, this data is not sufficient to estimate its global effect on cost-effectiveness; that's why we studied the whole direct costs generated by schizophrenia, the clinical effectiveness as well as the impact of therapeutics on the patients' quality of life. This work is a retrospective study of cost-effectiveness of clozapine treatment within three french psychiatric hospitals. The study was divided into three parts: - a twelve month period concerning the treatment before clozapine; - two twelve-month periods for the analysis of clozapine treatment. The clinical evaluation was based on the CGI and an autoevaluation scale. Whether schizophrenic patients were hospitalized or not and the importance of neuroleptic-associated medicines were taken into account to evaluate the Quality of Life. The economical assessment is represented by adding pharmaceutical costs, biological examinations costs and hospitalization costs. Thirty-seven patients, out of seventy-five, were being taken into account (26 men and 11 women). The results of the CGI and autoevaluation scale showed a global improvement in the pathology within the first year of treatment, which is confirmed during the second year. A statistically significant decrease in the length of full-time hospitalization was noticed; this profits to half-time hospitalization, indeed purely ambulatory. The direct costs per patient and per day insignificantly decreased between the first and the second period; on the other hand, the drop was statistically significant between the second and the third period. This study, based on a small amount of patients shows that the use of clozapine, which clearly helped the patients to improve, leads to a drop in global direct costs per patient, despite the high cost of the medicine itself, as well as an improvement in the quality of life. PMID- 9417403 TI - [Clozapine and treatment of negative symptoms]. AB - The efficacy of conventional neuroleptics in the treatment of the negative symptoms of schizophrenia is highly controversial. Clozapine, the leading atypical neuroleptic, has been shown to be effective, in the majority of the controlled studies, on both the positive and negative symptoms of the disease. It appears to be active vis-a-vis both the primary and secondary negative symptoms. However, several weeks (at least 12) are required for the effect on primary negative symptoms to clearly emerge. The position of clozapine in the treatment of patients presenting with a state of deficiency remains more debatable. PMID- 9417404 TI - Anti-neutrophil cytoplasmic antibodies and the endothelium many ways to vasculitis. PMID- 9417405 TI - The report of the Digestive Health InitiativeSM International Update Conference on Helicobacter pylori. PMID- 9417406 TI - [Controlling gastrointestinal risks in Bechterew disease. Interview by Dr. med. Katrina Recker]. PMID- 9417407 TI - [Orthopedic therapy in rheumatic diseases]. AB - The treatment of rheumatic disease requires an interdisciplinary approach in which the orthopedic surgeon is also involved. His contribution includes both conservative and operative procedures. Early synovectomy applied to joints and tendons is considered a preventive measure. Arthroscopic synovectomy belongs in the hands of an experienced operator. Joints that are not directly involved in weight-bearing can be treated by means of arthroplastic procedures, provided the cooperation of the patient is ensured. Arthrodesis is used preferentially to treat articular destruction of the ankle joint, the metacarpophalangeal joint of the thumb and the middle joints of the finger. Osteoporosis represents a relative contraindication. The treatment of rheumatic diseases of the joints using endoprostheses produces good primary results. The rate of loosening is not significantly increased, but late infection is somewhat higher than in other forms of articular disease. PMID- 9417408 TI - [Vitamin E in active arthroses and chronic polyarthritis. What is the value of alpha-tocopherol in therapy?]. AB - Known and recognized mechanisms of action of alpha-tocopherols are the grounds for the use of vitamin E in activated arthrosis or rheumatoid arthritis. This is also supported by the positive clinical results obtained in double-blind studies. On this basis, a change in the evaluation of these substances has taken place in recent years. Initially characterised as "unorthodox", "unconventional" or "additive" therapy, the term now applied tends to be "adjuvant-accepted" therapy. The results of double-blind studies and clinical empiricism support the following hyperthesis: The pathogenetic substrate "free oxygen radicals" increases quantitatively from activated arthrosis to (bland, mild) chronic polyarthritis. This could explain the graded differentiated antioxidative (or hypothetically the "central-analgesic") effects of alpha-tocopherols. While there still is no causal treatment with the alpha-tocopherols-pain alleviation and anti-inflammatory effect-are similar to those achieved with non-steroidal antiinflammatory drugs, the potency of which, however, is superior. PMID- 9417409 TI - [The clinical concept of spondyloarthropathies]. PMID- 9417411 TI - [Learning to live with rheumatism. Interview by Dr. rer. nat. Katharina Arnheim]. PMID- 9417410 TI - [Diseases of rheumatic origin. Evaluation in chronic recurrent course complicating therapy. Naturopathy Series, 15: Inflammatory systemic diseases]. PMID- 9417412 TI - [Chronic hepatitis. 6: Therapy of chronic hepatitis B and D]. PMID- 9417413 TI - [Common ignorance in tinnitus therapy. Reader opinion regarding critical views on tinnitus therapy]. PMID- 9417414 TI - [Antihypertensive therapy: the end of the flag pole?]. PMID- 9417415 TI - [New antihypertensive drugs: reasons for hope]. PMID- 9417416 TI - [Structured education for hypertensive patients. Interview by Dr. rer. nat. T. U. Keil]. PMID- 9417417 TI - [General practice 2000--what should be done today? The "health practitioner" is threatened by legal concerns]. PMID- 9417418 TI - [Physicians today--far removed from wealth. Nimbus and reality with reference to a survey]. PMID- 9417419 TI - [General health insurance patient "a little private". Cost measures versus chip card--details are still up in the air]. PMID- 9417420 TI - [A recall system for your type II diabetic patients. BDA-manual should improved standardized diagnosis and therapy for established physicians]. PMID- 9417421 TI - [More than a methadone treatment clinic]. PMID- 9417422 TI - [Phenomenology of hysterical pseudopsychoses]. AB - Clinical observation shows that hysteria can produce states that are difficult to differentiate from all forms of endogenous psychoses. Using phenomenological methodology, the elements and qualities of these hysterical states are analysed and described. It is demonstrated that the phenomena are properties of hysterical personalities, intensified and pronounced, but nevertheless intra- and extrapsychically targeted as "useful", i.e. serving a purpose. Phenomena that would be characteristic of real psychoses, are never seen. There is no "deprivation" of ego poser, and structural deformations are always absent. Considering the phenomenologico-psychopathological definitions of "psychosis", these hysterical states must be considered as pseudopsychoses. PMID- 9417423 TI - [Akathisia]. AB - The syndrome of akathisia typically consists of a subjective component, e.g. inner restlessness and an urge to move, and observable symptoms such as restless legs and inability to sit still. In most cases akathisia is caused by neuroleptics. There are several subtypes of akathisia according to the time of onset in the course of neuroleptic treatment. In clinical routine extrapyramidal motor disturbances are often underestimated or misinterpreted. As far as akathisia is concerned, differential diagnosis of restlessness or of repetitive movement patterns may be problematic. Non-compliance and impulsive behaviour are regarded as possible complications of akathisia, but systematic investigations are lacking. The pathophysiology of akathisia is not clear, but it probably differs from other pharmacologically induced motor disturbances. If warrantable, the first step in akathisia treatment is dose-reduction of the causing agent. Anticholinergic drugs, benzodiazepines, and beta-receptor blockers may be effective. Clinical assessment and survey of the patient's behaviour, e.g. during occupational therapy and group therapy is important for an early diagnosis of akathisia so that complications may be minimised. PMID- 9417424 TI - [New possibilities in therapy and rehabilitation of alcohol dependent patients. Catamnestic study of the efficacy of ambulatory withdrawal therapy exemplified by a model facility]. AB - We report on catamnestic and clinical results of a 18-24 month follow-up study of 65 alcohol-dependent patients who in 1992-1994 took part in an 8-month outpatient treatment programme. The psychotherapeutic concept of this treatment facility is described. 51 of the 65 patients who had participated in the programme could be subsequently personally interviewed, 7 patients refused to take part, 6 could not be reached and 1 had died. 40 of the 51 patients had properly completed the outpatient treatment. Assuming that all patients who could not be interviewed or refused, were relapsers the abstinence rate was found to be 48% (n = 31). Although selective factors may contribute to this result, the clinical findings so far seem to indicate that outpatient treatment for alcoholics is a promising new therapeutic approach in the treatment of alcoholism. Further studies are needed to assess the possible benefits and the indications for this outpatient treatment in greater detail. PMID- 9417425 TI - [Wernicke-Korsakow syndrome: clinical aspects, pathophysiology and therapeutic approaches]. AB - In ICD-10 or DSM IV, the Wernicke-Korsakow Syndrome (WKS) is mentioned among the criteria for the alcohol amnestic disorder. Rarely seen in clinical practice, the Wernicke-Korsakow Syndrome is important in differential diagnosis of amnestic syndromes, because life-threatening conditions can occur. A specific treatment is available for the acute form of Wernicke encephalopathy, in many cases a Wernicke encephalopathy shows a progression to the Korsakow-Syndrome, which is the chronic form of this disease. The aim of this article is to review recent developments in clinical, etiological and pathophysiological research of WKS. The role of thiamin dependent enzymes like transketolase and the importance of the NMDA (N-Methyl-D Aspartate) System is discussed. New trends in diagnostics are shown, as are differential diagnosis and epidemiology. Finally, the rarely investigated drug therapy of the Korsakow syndrome with clonidine and fluvoxamine is reviewed. PMID- 9417426 TI - [Sleep apnea syndrome and cerebral lesions--a prospective MRI study]. AB - It seems well known that sleep apnea syndrome (SAS) is associated with cardiovascular complications, inclusive myocardial infarctions. Our study aimed to test the hypothesis that patients suffering from cerebral SAS have more cerebrovascular findings resp. brain infarctions than a matched control group. We analysed prospectively MRI studies of 14 patients with SAS and controls without SAS in respect of specific and nonspecific vascular cerebral lesions including atrophic changes of the brain. In contrast to our expectations, the rate of SAS associated ischemic brain lesions is unimportant and not significantly different from age-associated brain lesions in controls. The rate of brain infarctions is distinctly different from that of myocardial infarctions. The outcomes are discussed with regard to new results of "ischemia-induced" ischemic tolerance on different tissues. PMID- 9417427 TI - [Antithrombotic therapy after cerebral ischemia]. AB - Following cerebral ischaemia a recurrent stroke must be avoided in most patients by means of antithrombotic agents. Based on the results reviewed here of new therapy studies, we discuss the presently available antithrombotic treatment options for prophylaxis in ischaemic stroke. TASS (Ticlopidine Aspirin Stroke Study) and CATS (Canadian American Ticlopidine Study) are two multicentre studies investigating the effect of ticlopidine, a new antiplatelet agent of the thienopyridine family, compared to acetylsalicylic acid (ASA) respectively placebo, in the secondary prophylaxis of ischaemic stroke. A significant relative risk reduction of ticlopidine against ASA (21%) and against placebo (28.1%) was shown. CAPRIE (Clopidogrel vs. Aspirin in Patients with Risk of Ischemic Events) evaluated clopidogrel and ASA in the secondary prophylaxe of stroke, myocardial infarction and peripheral vascular occlusive disease. Clopidogrel has been shown to be as effective as ticlopidine compared to ASA in the secondary prevention of vascular disease but had the advantage of a far less severe side effect profile as ticlopidine. ESPS 2 (2nd European Stroke Prevention Study) compared dipyridamole and ASA alone and in combination against placebo in stroke prevention. The combination of agents showed a 24.4% relative risk reduction to suffer ischaemic stroke as opposed to placebo. The ranking of heparin and heparinoids in the secondary prevention of ischaemic stroke has not been completely established but seems to diminish according to recently published data from three major trials. The American TOAST study (Trial of Org 10172 in Acute Stroke Treatment) failed to prove any advantage of intravenous Orgaran compared to placebo. In IST (International Stroke Trial) and CAST (Chinese Acute Stroke Trial) the benefits of heparin are invalidated by a higher bleeding rate of patients on intravenous heparin therapy. Furthermore, the results of IST have to be judged critically because of significant methodical inadequacies. When applying antithrombotic agents, therapeutic effect and presumed better outcome should be weighed against the risk of associated bleedings. The indication for an antithrombotic treatment should be reevaluated in regular control examinations and the possibility of a less aggressive treatment should be considered. PMID- 9417429 TI - [Reflux-associated diseases of the upper respiratory tract]. PMID- 9417428 TI - [The effectiveness of antibiotics in chronic middle ear effusion and chronic suppurative otitis media. Comment on 2 contributions from Lancet]. PMID- 9417430 TI - [Ototoxicity of aminoglycoside antibiotics: an alternative in sight?]. PMID- 9417431 TI - [Follow-up of caloric response after acute peripheral dysfunction]. PMID- 9417432 TI - [Guidelines/algorithms of the German Society of Otorhinolaryngology, Head and Neck Surgery]. PMID- 9417433 TI - [Long-term follow-up of fronto-basal dura-plasty]. AB - A safe closure of a dura lesion is necessary on account of the risk of potentially fatal (late) meningitis. 161 duraplasties of the frontal skull base carried out from 1979 to 1994 at the ENT-department Fulda were evaluated in a retrospective study in regard to etiology, operative techniques and results. Duraplasty of the rhinobasis was indicated in 70 cases of rhinobasal trauma, 47 cases after paranasal sinus surgery, 36 cases of tumors and 8 malformations. After an average follow-up time of 6 years the patients were interviewed for postoperative liquorrhea, sinusitis treated with antibiotics and meningitis. As an objective measure to verify the tight closure of the treated CSF-leaks a fluorescein test was performed in 50.9% 6 to 8 weeks after the operation. Duraplasty was successful in more than 96%. The approach and technique to perform a duraplasty have to be chosen individually considering size, location and etiology of the dural defect. In the majority of dural defects in the area of the frontal skull base reconstruction can be carried out now a days via an endonasal approach. By use of allogenic tissue, a mucosal flap from the surrounding area to cover the graft and fibrin clue good results were obtained. PMID- 9417434 TI - [Follow-up of caloric test response after acute peripheral vestibular dysfunction]. AB - This study examined retrospectively the spontaneous recovery of patients with an acute peripheral vestibular deficit in order to determine whether the caloric test response and with it vestibular function improves over time. The caloric bithermal was tested three times on 79 patients who were hospitalised with an acute deficit. The first test was recorded on emergency admission by observing nystagmus beats under the Frenzel glasses. Two to five days later a complete electronystagmus (ENG) examination was performed. A second ENG was performed, on average, 4 months later. 46% of the patients recovered a normal caloric canal paresis value (less than 32%). By comparing the canal paresis values in the first and second ENG an improvement exceeding 30% was demonstrated in 50% of the patients and there was no correlation between the extent of the canal paresis deficit and the amount of recovery. A simultaneous cochlear deficit had no influence on the recovery of vestibular function. PMID- 9417435 TI - [Late sequelae of lateral and central mid-facial fractures after osteosynthesis with miniplates]. AB - Despite significant progress in the therapy of craniofacial injuries permanent consequences of the trauma are still likely to occur. It has been possible to follow-up 105 (65.2%) of the 161 patients who were treated for lateral and central midfacial fractures in our department between 1989 and 1991. Pathological findings on follow-up were found in 48.6% of our patients. Most frequent were persistent disturbances of the sensitivity in the area of the second branch of the trigeminal nerve (32.4%). Patients who were operated within the first week after trauma did have significantly less sensory disturbances than patients who underwent surgery later. 7.6% of our patients had permanent double vision (4.8% in their primary vision field). After subciliary incision 3% had an ectropium which was not found after transconjunctival incision. There was no significant difference in the cosmetic outcome of the two approaches. In one patient who received silicone for restoration of the orbital floor the plate did migrate. Other materials that were used for orbital floor reconstruction did not cause problems. Based on the post-operative results improvements for the therapeutic regimen are discussed. PMID- 9417436 TI - [New methods of type II tympanoplasty in erosion of the long incus process]. AB - Reliable methods of reconstruction of the ossicular chain in the situation of an isolated errosion of the long process of the incus using a tympanoplasty type II have not been available until recently. Instead, the tympanoplasty type III has been generally performed with the interposition of an autologous incus. In this presentation, we are describing two methods for reconstruction of the ossicular chain between the in-situ residing incus and the stapes on the other side so that the direct connection eventually will result in a tympanoplasty type II. In the first case, we used ionomeric cement in a way that features two characteristics: the direct connection between the stapes and the long process of the incus could be achieved as well as an articulation that was created on the head of the stapes. Hence, a too stiff connection between the head of the stapes and the long process of the incus could be avoided. In addition, a new method for precise microapplication of cooled bone cement (IONOCAP LV) with a syringe will be presented. In the second method titanium-gold-angle prostheses have been crimped to the long process of the incus and positioned onto the head of the stapes in the way of an articulation. So far, comparison of the audiological results of those two methods of a tympanoplasty type II reveal in average better results than postoperative conductive hearing thresholds of the conventional tympanoplasty type III. If the achieved results can be reproduced on a larger number of patients, the expected audiological results are likely to resemble those of stapes surgery. PMID- 9417437 TI - [Polarographic detection of reoxygenation of lymph node metastases in the initial phase of primary radio- and radiochemotherapy in patients with advanced squamous epithelial carcinomas of the head and neck]. AB - The therapeutic effect of radiotherapy depends amongst other things on the degree of oxygenation of tumour tissue. Epithelial carcinomas of the head-neck region exhibit considerable hypoxic areas which vary markedly between individuals and not encompassed by current staging and grading. It is assumed that during radiation treatment a reoxygenation of hypoxic tumour cells takes place. It was investigated wether the occurrence of reoxygenation could be determined by invasive oxygen partial pressure measurements in lymph node metastases. Using a needle probe inserted transcutaneously into a lymph node polarographic oxygen determinations (Eppendorf pO2-Histiograph) were made on 13 patients with advanced oro- and hypopharyngeal carcinomas before therapy and after a week of accelerated radio- or radiochemotherapy. Low pO2 values before treatment (median 13.5 mmHg, average 20.3 mmHg) and a hypoxic fraction (pO2 < 10 mmHg) of 45.2% indicated manifest tumour hypoxia. After the first week of treatment a significant increase in the median- (24.1 mmHg) and the average pO2 (28.2 mmHg) as well as a reduction in the hypoxic fraction (27%) were observed. Invasive pO2 histiography fulfils the requirements for a method to confirm tumour hypoxia in head-neck tumours. The results obtained indicate that reoxygenation occurs during the initial phases of radio- and radio-chemotherapy. PMID- 9417438 TI - [Hearing tests in extended high frequency range in pre-school age children. Initial results]. AB - Since reliable headphones are now available, clinical audiometry can also be performed in extended high frequencies (EHF). Hearing in frequencies over 10 kHz is more influenced by age, noise and toxicity. Thus it is useful to take additionally results in children to establish the normal hearing threshold. In the present study, 35 pre-school children (ages 4-7 years) were tested by EHF (8 kHz-16 kHz) using a new Sennheiser HDA-200 headphone. The hearing thresholds recorded corresponded to those of other studied. Median and standard deviations for 10 kHz were 25 dB(SPL) +/- 12 dB(SPL), 35 dB(SPL) +/- 12 dB(SPL) for 12.5 kHz and 50 dB(SPL) +/- 15 dB(SPL) for 16 kHz. In the older children (> 5 years), hearing thresholds could be more reliably determined and were 10 dB(SPL) better than in the younger children (< 5 years). Thresholds at 10 kHz-12.5 kHz corresponded to those found in adults, but were more sensitive by 5 dB(SPL)-15 dB(SPL) in the 14 kHz-16 kHz range. PMID- 9417439 TI - [Axillary metastases of oropharyngeal carcinoma after pectoralis major flap]. AB - Reconstruction of defects after resection of advanced head and neck tumours with the pedicled myocutaneous pectoralis-major-flap is a well established method of regional plastic surgery. A 66 year old female patient with an oropharyngeal squamous cell carcinoma was treated by surgery and radiotherapy in curative intention. A few months later a local recurrence could be resected by a lateral pharyngotomy while the defect was reconstructed with a myocutaneous pectoralis major-flap. Together with another local relapse at the pectoralis-major-flap in the oropharynx a painless tough swelling developed in the ipsilateral axilla which revealed histologically as a lymph node metastasis of a squamous cell carcinoma. Because no second malignoma as a source for this metastasis could be found, a lymphogenous spread of the oropharyngeal carcinoma along the pedicle of the flap could be the most probable explanation for this rare event. As a conclusion of this observation all patients with pedicled reconstructions after resection of head and neck cancer should not only be examined in the tributary lymph node levels of the neck but also in the axilla during oncological follow up. PMID- 9417440 TI - [Progressive unilateral exophthalmos. Aspergillosis of the orbits]. PMID- 9417441 TI - [Therapy study. Comparison of postoperative irradiation versus omission of irradiation in mouth cavity and supraglottic carcinomas]. PMID- 9417442 TI - [Drug treatment of tumor pain]. PMID- 9417443 TI - [Clinical significance of tumor suppressor gene p53 in head and neck tumors]. PMID- 9417444 TI - [Angelman syndrome: genetic defect with delayed language development]. PMID- 9417445 TI - [Retraining therapy in tinnitus. Paradigm change or old wine in new bottles?]. PMID- 9417446 TI - [Sleep medicine]. PMID- 9417447 TI - [Guidelines/algorithms of the German Society of Otorhinolaryngology, head and neck surgery. German Society of Otorhinolaryngology, Head and Neck Surgery]. PMID- 9417448 TI - [Treatment of recurrent epistaxis in Rendu-Osler-Weber disease]. AB - Rendu-Osler-Weber disease is an autosomally dominant inherited disease which affects the connective tissue of blood vessels. The major symptom is recurrent epistaxis for which still exists no generally recommended treatment. Treatment modalities discussed in the literature include bilateral closure of the nostrils, brachytherapy, vessel embolization, hormone therapy, dermoplasties, as well as different kinds of laser therapies. Advantages and disadvantages of single treatment modalities are discussed. Dermoplasty and laser therapy seem to be the most suitable treatment procedures currently available for recurrent epistaxis in affected patients. Treatment carried out with the argon, KTP or ND:YAG laser has proven to be suitable as initial therapy. In most cases the additional use of long-term applications of a soft nasal ointment allows a lasting successful treatment. Our experiences show that dermoplasty should be reserved for cases with continued bleeding despite laser treatment but only rarely offers a long term cure in the sense of a total recovery from epistaxis. PMID- 9417449 TI - [Erosion and necrosis of the long process of the incus after otosclerosis operation]. AB - The frequency and possible causes of erosion and necrosis of the long process of the incus induced by various types of prostheses following stapes surgery for otosclerosis are discussed. Between 1987 and 1995, 1144 stapes operations were performed including 117 revisions of stapes surgery performed by third parties and 27 revisions performed by the author. For the first time, necrosis of the long process of the incus caused by teflonplatinum piston is described on the basis of findings in five cases, in which the platinum loop had completely transmigrated the incudal process in a medial direction. By using pictures of the teflon-platinum piston and a gold piston taken by a scanning electron microscopy and a comparison between the use of a teflon-wire piston and a gold piston in two operations, an hypothesis is made that the winding grooves in the surface of the teflon shaft together with impaired middle ear mechanics were the main causes of a scar being contracted in the direction of the oval window niche with resulting necrosis of the incudal process. Requirements for an optimum stapes prosthesis are summarized as a result of the operative observations made. PMID- 9417450 TI - [Effectiveness of partial and complete instrumental masking in chronic tinnitus. Studies with reference to retraining therapy]. AB - Jastreboff und Hazell [9] developed a neurophysiological approach to tinnitus perception, including the important role of the central nervous system in the maintenance and intrusiveness of tinnitus. They introduced tinnitus-retraining therapy, consisting of four different strategies: (1) directive and person centered counseling; (2) hearing aids and/or noise generators and/or environmental sounds; (3) psychological therapy; (4) adjacent therapies. Tinnitus should not be masked as with a tinnitus-masker, but must be able to be heard in addition to the noise! A noise generator or hearing aid should be worn at least 6 8 h per day over a period of up to 18 months. In additions several clinical visits are required in order to reinforce the counseling. The actual results show complete tinnitus remission for about 20-30% and partial remission for 50-60% of the patients [6]. We report on a retrospective study in patients wearing hearing aids or tinnitus-maskers over a period of 3 years. We compared the results of patients using partial tinnitus masking to those using complete masking. The tinnitus-related and general psychological complaints were acquired by the 52 item tinnitus questionnaire developed by Hallam et al. [4] and modified by Goebel and Hiller [3]. To describe the dimensions of tinnitus-related distress the scales are labelled emotional distress, cognitive distress, emotional and cognitive distress, intrusiveness, auditory perceptual difficulties, sleep disturbance and somatic complaints. Positive changes for the global tinnitus questionnaire score of more than 10 points are significant in the dimensions of tinnitus-related distress and are described as partial tinnitus-reduction. The group with partial masking effects can be compared to those performing retraining therapy to day because directive and personal centered counseling were integrated for all patients. Patients reporting partial masking effects through their aids (hearing aid or noise generator) showed more effective treatment results (reduced or disappeared tinnitus) than those using complete masking effects (P < 0.05). The reported results are improved by current investigations in Germany showing about 20-30% tinnitus remission and 50-60% significant reduction after 1 year of treatment [1]. Further scientific investigations must be carried out to evaluate the appropriateness and effectiveness of the retraining therapy and with regard to unique quality standards. PMID- 9417451 TI - [Chloride conduction of nasal fibroblasts in polyposis patients with cystic fibrosis and in patients without cystic fibrosis. Relevance for the ENT physician]. AB - Cystic fibrosis (CF) is a complex systemic disease that has pathological alterations in the upper airways, including the recurrent formation of nasal polyps. Although the fibroblast is the predominant cell type in nasal stroma and nasal polyps, little is known about the electrophysiological properties of nasal fibroblasts. We investigated whether fibroblasts possess a cAMP-regulated chloride conductance which is impaired in patients with CF. Thus far the few studies concerning conductance in fibroblasts have been performed on skin fibroblasts using indirect methods and have yielded conflicting results. Therefore we studied chloride conductance in fused nasal fibroblasts by employing conventional microelectrodes. We have demonstrated that a cAMP-regulated chloride conductance is present in fibroblasts. However, this chloride conductance cannot be activated in fibroblasts from CF-patients. Thus, we present direct evidence that the impairment of the cAMP-regulated chloride conductance in CF is not confined to epithelial cells but also affects the fibroblast. We discuss how this conductance might modulate fibroblast proliferation to produce polyp formation. PMID- 9417452 TI - [99mTc-MDP-SPECT for detection of subclinical mandibular infiltration of squamous epithelial carcinoma]. AB - A prospective study was performed to compare the sensitivity and accuracy of ultrasound (US), computed tomography (CT), and single photon emission computed tomography (SPECT) as a bone scan to detect subclinical invasion of the mandible by squamous cell carcinoma of the oral cavity and floor of mouth. METHODS: The present study reports data on 35 patients with squamous cell carcinoma of the oral cavity. SPECT, mandibular orthopantomography (OPG), US, CT and magnetic resonance imaging (MRI) were performed as well as histologic examination of resected mandibles. Transverse slices of 12 mm thickness were reconstructed for SPECT and semi-quantitative assessment of radionuclide uptake was performed with a scoring system based on comparisons of tracer uptake in the mandible to that in an unaffected part of the mandible. Lesion to non-lesion count ratios in the mandible in 3 h SPECT images were calculated. OPG, US and CT were evaluated for comparison. RESULTS: In 12/35 patients, histologic examination showed carcinoma infiltrating mandibular bone. SPECT correctly predicted mandibular invasion in 11/12 cases, with no false-positive results. CT detected correctly 3/12 lesions, while OPG and ultrasound each demonstrated 2/12 tumors. CONCLUSION: Bone SPECT was found to provide an accurate means for assessing tumor invasion to the mandible by intraoral squamous cell carcinoma. As a complementary study to one of the other imaging modalities it was shown to have a high specificity, indicating that it should be performed routinely in patients with suspect mandibular bone involvement by causes. PMID- 9417453 TI - [Indistinguishable space-occupying lesion of the palate. Neurinoma of the palate]. PMID- 9417454 TI - [Functional voice quality assessment in curative microsurgery of laryngeal malignancies. Postoperative voice rehabilitation based on the "laryngeal double valve function"]. AB - According to Negus and Pressman the sphincter systems of the vocal folds and the ventricular folds form a respiratory "laryngeal double valve function". Correspondingly, we found a physiological phonation system of the glottis and a pathological-compensatory one of the supraglottis. They appear to be regulated through an automatic phonatory control system with the glottal phonatory function evidently acting as sensor level. In order to confirm this hypothesis, objective voice analyses with glottal-relevant parameters of 26 voice-rehabilitated patients after minimally invasive laser surgery of glottal carcinomas are presented and integrated into a "hoarseness diagram" with the coordinates roughness and breathiness. Using statistically deliminated acoustic dusters, our data show a qualitative hierarchy of different postoperative phonation mechanisms. They demonstrate the influence of the vibratory capacity of glottal and supraglottal structures on the quality of the vibratory closure. Both functional parameters evidently determine the resulting voice quality in the sense of our hypothesis. PMID- 9417455 TI - [Isolated histiocytosis X as rare differential diagnosis of indistinguishable neck lymph node swelling. A case report]. AB - Cervical lymphadenopathies are widespread diseases in otorhinolaryngology. There are many differential diagnoses which have to be considered when swelling persists. To gain further information it is advisable to perform sonographically controlled fine needle aspiration biopsy (FNP). If there is no reliable result, it is necessary to remove a lymph node. We report on the rare case of isolated histiocytosis X in lymph nodes. A 63-year-old male had noticed a swelling of the right side of the neck about 6 weeks previously. The preliminary examinations (FNP/blood examinations) revealed no pathological findings. Only the histological examination delivered the diagnosis of histiocytosis X. Systemic manifestation was excluded by staging examinations. No further therapy was necessary after lymph node extirpation. The report demonstrates how to diagnose and treat this rare disease. PMID- 9417456 TI - [Endoscopically controlled dilatative puncture tracheostomy]. PMID- 9417457 TI - [Pregnancy in women with a mechanical heart valve. Review of the literature]. AB - AIM: To define therapeutic ways to manage obstetrics at an optimal level for a patient carrying a mechanical prosthetic heart valve. METHOD: From a review of literature and documented cases, we propose an obstetrical and cardiological management scheme for pregnant patients carrying a mechanical prosthetic heart valve. We deal successively with the preventive (anticoagulation) and curative treatment (surgery and thrombolytic therapy) of valvular thrombosis, in fact, a severe -but unfortunately frequent- complication of these pregnancies. RESULTS: Pregnancy concerning patients with mechanical prosthetic heart valves is a high risk pregnancy. In fact, the risk of thrombo-embolic accidents even with a closely followed anticoagulant treatment seems to come from the existence of the prothese, the state of physiological hypercoagulation and peri-partum hemorrhages. Naturally, multidisciplinary follow-up is indispensable. The prescription of anticoagulant treatments must respect the main principals and the normal counter-indications in order to minimise maternal and fetal complications. CONCLUSION: Pregnancy amongst patients carrying mechanical prosthetic heart valves should be considered rare and highly exceptional cases, from a cardio vascular surgeon's point of view. For obstetricians, it is sometimes difficult to forbid pregnancy to a nullipara. The discussion whether to authorize a pregnancy will be treated case-by-case, holding into account the socio-cultural environment of the patient, as the principles of an anticoagulant treatment and the underlying risks must be well understood. PMID- 9417458 TI - [Circulation and cerebral metabolism in neonatal hypoxia-ischemia]. AB - The basic physiological variable in hypoxic-ischaemic brain injury is cerebral oxygen delivery. When oxygen delivery becomes insufficient to meet the cellular demands for oxygen, a sequence of biochemical events will be triggered leading to cell death. High levels of CBF following severe birth asphyxia is now well documented by Doppler ultrasound which has been shown to be a useful prognostic indicator following birth asphyxia. Near infrared spectroscopy (NIRS) is of great potential value since it may be used at the bed-sid and allows to measure the cerebral blood volume and the concentrations of cytochrome aa3. Magnetic resonance spectroscopy (MRS) allows noninvasive assessment of cerebral metabolism in asphyxiated neonates. 31P MRS has demonstrated that birth asphyxia leads to delayed impairment of cerebral energy metabolism and is predictive of later neurodevelopmental outcome. 1H MRS has shown lactate accumulation and a later decline in N-acetyl aspartate concentration. PMID- 9417459 TI - [Mass screening programs for breast cancer in France. Comparative evaluation]. AB - OBJECTIVE: The purpose of this work was to comparatively assess the results of mass screening programs for breast cancer implemented in six French departments in 1986, within the scope of the National Fund for Health Prevention, Education and Information of the National Health Insurance Office of Salaried Workers. MATERIAL AND METHODS: The data collected by the screening centres were analyzed by ten assessment teams that were independent from the program promotion staff, all using the same evaluation form. A complementary population study performed in eight French districts then, allowed assessing the frequency of self-referred screening (mammography performed out of program). RESULTS: The rate of participation in screening programs, in relation to the invited population, ranged from 21 to 48%, according to the district (36% in average). This low participation was probably related to the extent of self-referred screening. In fact, 19 to 40% of women, according to the district, had previously had a screening mammographic coverage: rate was around 68% in women aged 50 to 69 years. Positive findings with mammography ranged from 4.5 to 15.8% (10.1% in average), while intervention rates ranged from 0.7 to 1.6% and detection rates from 3.8 to 6.2%. The ratio between benign tumors and cancers ranged from 0.7 to 2.1 according to the district. In order to enlighten the judgement on French results, we propose a comparison with the international standards in force. CONCLUSION: The various experiences with breast cancer screening in France show that this screening is technically feasible on the basis of existing medical structures. However, some criteria are still below the expected values, especially if compared with international standards. This result is probably accounted for by the high rate self-referred screening before age 40 in France. In these conditions, the question is whether extending breast cancer screening programs in France is an appropriate course of action. PMID- 9417460 TI - [Cesarean sections in France: impact of organizational factors on different utilization rates]. AB - In this study, we analysed the potential impact of organizational factors to explain the variation of cesarean sections' rates. We used a retrospective sample of 84,372 deliveries and two subsamples of low risk deliveries for cesarean sections. We determined different organisational factors that included: juridical and financial status of maternities, their architecture, the type of on-call for obstetricians, pediatrists and anesthetists, the annual number of deliveries and the level of pediatric staff and equipments of the maternities. We used multiple regression techniques to study the specific effect of each parameter, while controlling effects of age and parity of the mothers. We have found that even on the low risk samples, variation of rates were important. The type of on-call, the level of pediatric services and the architecture of maternities exerted a strong and significant effect on the rate of cesarean sections compared to the absence of impact of the number of deliveries. We discuss the reasons why, explaining the occurrence of those factors and then, stress the need to take into account the relevant factors for organizational audits. It appears that, in the context of the new regulation of the health system, these results should give obstetricians reasons to enhance their efforts to correct inefficient practices and to respect consensual guidelines and joint accreditation of obstetric and pediatric units. PMID- 9417461 TI - [CIN and pregnancy. Apropos of 16 cases and review of the literature]. AB - AIM: To assess the means to diagnose grade 3 cervical intraepithelial neoplasia (CIN 3) during pregnancy, with special consideration to the risk of overlooking invasive lesions. STUDY DESIGN: a retrospective study on 16 cases of CIN 3 over 4 years and a literature review. RESULTS: Smear tests were indicative of low-grade lesions in 8 cases. Colposcopy always was suggestive and accompanied by biopsy. No conisations were performed during pregnancy but one was performed after delivery in 15 of 16 cases. After delivery, comparing conisation results with those of recent biopsies revealed that some lesions were less severe and others more severe than during pregnancy. So, a CIN 3 and a microinvasive carcinoma observed during pregnancy changed into CIN 2 and CIN 3, respectively, after delivery. In contrast, two pregnancy CIN 2 were seen as CIN 3 in post-partum and three CIN 3 as MIC. Such variations have been described in the literature and have several causes. In particular, improvements may sometimes reach "normalisation" in post partum although, recurrences remain possible. CONCLUSIONS: when the smear tests performed during pregnancy are indicative of cervical intra-epithalial lesions, whatever the severity, colposcopy with biopsies are mandatory. If a CIN 3 is detected, early MIC should be removed, which may require specific treatment according to the invasive degree. Colposcopy with guided biopsies is a safe and reliable diagnostic means. After delivery, the cervix should always be reexamined, preferably by conisation, even if smears or colposcopy were normal. PMID- 9417462 TI - [Does axillary liposuction modify histologic study of excised lymph nodes?]. AB - OBJECTIVE: To determine the pathological features of lymph nodes removed by axillary liposuction. METHODS: A prospective study of 34 axillary dissections performed from July 1995 to September 1996 in patients with breast cancer N0. After lipolysis, the fat was drained from the axillary cavity by means of liposuction (Karman nozzle ch.8; suction pressure of 1 Bar). The remaining nodes were removed by an endoscopic dissection in conservative treatments. The remaining nodes were removed during modified radical mastectomy in non conservative treatments. Lymph nodes were fixed in formol and examined by the pathologist. RESULTS: An average of 15 lymph nodes (8-35) were removed. 502 lymph nodes were examined: 458 (91%) were not involved and 44 (9%) were involved, including 21 (4%) with rupture of the capsule. No pathological traumatism was seen. CONCLUSION: Axillary liposuction did not alter the pathological features of lymph nodes in our study. PMID- 9417464 TI - [Echographic measurement of the inferior uterine segment for assessing the risk of uterine rupture]. AB - BACKGROUND: Ultrasonography has been used to examine the scarred uterus in women who have had previous cesarean sections in an attempt to assess the risk of rupture of the scar during subsequent labor. The predictive value of such measurements has not been adequately assessed, however. We aimed to evaluate the usefulness of sonographic measurement of the lower uterine segment before labor in predicting the risk of intrapartum uterine rupture. METHODS: In this prospective observational study, the obstetricians were not told the ultrasonographic findings and did not use them to make decisions about type of delivery. Eligible patients were those with previous cesarean sections booked for delivery at our hospital. 642 patients underwent ultrasound examination at 36-38 weeks' gestation, and were allocated to four groups according to the thickness of the lower uterine segment. Ultrasonographic findings were compared with those of physical examination at delivery. FINDINGS: The overall frequency of defective scars was 4.0% (15 uterine ruptures and 10 dehiscences). The frequency of defects rose as the thickness of the lower uterine segment decreased: there were no defects among 278 women with measurements greater than 4.5 mm, three (2%) among 177 patients with values of 3.6-4.5 mm, 14 (10%) among 136 patients with values of 2.6-3.5 mm, and eight (16%) among 51 women with values of 1.6-2.5 mm. With a cut-off value of 3.5 mm, the sensitivity of ultrasonographic measurement was 88.0%, the specificity 73.2%, positive predictive value 11.8%, and negative predictive value 99.3%. INTERPRETATION: Our results show that the risk of a defective scar is directly correlated to the degree of thinning of the lower uterine segment at around 37 weeks of pregnancy. The high negative predictive value of the method may encourage obstetricians in hospitals where routine repeat elective cesarean is the standard procedure to offer a trial of labor to patients with a thickness value of 3.5 mm or greater. PMID- 9417463 TI - [Celioscopic ovariectomy. Retrospective study of 56 ovariectomies]. AB - OBJECTIVE: To describe a technique of laparoscopic oophorectomy, and evaluate its feasibility, limits and complications. DESIGN: We performed a retrospective study of 34 patients who underwent laparoscopic oophorectomy, from 1 December 1992 to 28 February 1995. SETTING: Centre Hospitalier Intercommunal, Creteil, and Institut Gustave Roussy, Villejuif. SUBJECTS: Mean age was 58 years (range: 42 to 74 years). In post-menopausal patients with ovarian cyst, a systematic oophorectomy was performed (33 patients). Among post-menopausal women, a systematic contro-lateral oophorectomy was done in 77% of cases. Castration was preconized in one woman with previous breast carcinoma. TECHNIC: Removal of the ovary was performed by dessication and division of the utero-ovarian junction and of the mesovarium. RESULTS: Among the 34 women, 32 (94%) had an exclusive laparoscopic procedure, 2 had laparo-conversion. The reasons of laparo-conversion were the presence of adhesions in one case and presumption of ovarian malignancy (Border-line ovarian tumor) in the other case. For the 32 women with laparoscopic treatment, 22 (65%) had bilateral oophorectomy. Per-operative complication rate was 6.2% (an epigastric vessel injury in one case and an hemorrhage during laparoscopic adhesiolysis in an other case). The post-operative time was uneventful. CONCLUSION: This technique of laparoscopic oophorectomy is simple, rapid and has a low rate of per and post-operative complications. In addition, this technique has the advantage to prevent the risk of ureteral injury. PMID- 9417465 TI - [3-dimensional (3D) echography in obstetrics. Advantages and limits]. AB - Three-dimensional ultrasound (3D) offers new options in imaging modes: such as simultaneous rotation and translation of the three perpendicular planes displayed, surface rendering, or transparent mode providing an imaging of structures with high echogenicity (bones, skull). This new imaging mode is extending the field of conventional two-dimensional ultrasound, but for now it has to be evaluated. PMID- 9417467 TI - [Autosomal dominant polycystic kidney. Apropos of 2 cases. Review of the literature]. AB - Autosomal dominant polycystic kidney disease (ADPKD) is the most common hereditary disease. Genetic molecular methods can make the diagnosis of at least three different types of ADPKD. ADPKD concerns young people and complications such as hypertension and decreased renal function occur more frequently if onset is early, if it is a type 1, and if the patient is a woman. The pregnant woman with autosomal dominant cystic disease is at particular high risk of obstetrical complications. Prenatal diagnosis is possible. PMID- 9417466 TI - [Medical treatment exclusively for cervical pregnancy with in situ methotrexate]. AB - We present a case report of cervical pregnancy with medical treatment. Medical treatment consisted in injection of methotrexate (50 mg) into the pregnancy, on the first, third and seventh day. Ultrasound and Doppler give important information for follow up. The pregnancy totally resolved and the patient did not need any further treatment. PMID- 9417468 TI - [A case of neonatal Meckel's diverticulum with prenatal ultrasound diagnosis]. AB - We report one case of cystic Meckel's diverticulum detected by fetal ultrasound echography with an anechogen image during the third trimester of pregnancy. The interest of this antenatal diagnose is to quickly manage surgical care at birth. PMID- 9417469 TI - [Ultrasound abnormalities of the placenta during triploidy with pre-eclampsia in the 2nd trimester]. AB - A documented case of triploidism which developed to 20 weeks gestation underlines the importance of early ultrasonographic assessment of the trophoblast. Large placentas, especially when pre-eclampsia occurs, should lead to assay of beta-hCG and search for chromosome anomalies. Paternal genomic origin was suggested based on the large volume of the placenta. PMID- 9417470 TI - [Dissection of the ascending aorta in pregnancy. Apropos of a case and review of the literature]. AB - Dissection of ascendant aorta is infrequent during pregnancy. Given this low prevalence, diagnosis is often delayed or even unrecognized. This delay in diagnosis can have a highly jeopardizing effect on vital maternal prognosis. In this study, we report a case of ascendant aorta dissection in the 3rd trimester of pregnancy with maternal and fetal survival through combined surgery of a cesarean section and replacement of the ascendant aorta. PMID- 9417471 TI - [Apropos of evaluating the risk of uterine rupture by ultrasonic assessment of the inferior uterine segment]. PMID- 9417472 TI - Interleukin-12, dendritic cells, and the initiation of host-protective mechanisms against Toxoplasma gondii. PMID- 9417475 TI - [A plea for mandatory magnetic resonance imaging]. PMID- 9417474 TI - Special issue: Cellular and molecular membrane transport--a symposium in honor of Arnost Kleinzeller. Salsbury Cove, Maine, August 1-3, 1996. PMID- 9417473 TI - What goes up must come down: the emerging spectrum of inhibitory receptors. PMID- 9417476 TI - [What is your diagnosis? Mycotic aspergillus sinusitis]. PMID- 9417477 TI - [Identification of the central sulcus using the scanner and MRI]. AB - Methods to directly and indirectly identify the central sulcus are presented. In the axial plan, direct method is remarkable but obviously requires good visualization of the sulci in the central region. Sulci are readily visible in 90% of the cases on CT scans and in 50% of the cases on MRI. The method can also be applied when tumoral development erases the cerebral sulci by direct lecture of the controlateral rolandic region and right-left transfer. Within the precision limits of the method, it can be considered that the central sulci are symmetrical. The main signs are: the relative morphologies of the superior frontal sulcus and the precentral sulcus, the hook-shaped aspect of the middle part of the central sulcus, the internal end of the central sulcus projection anteriorly to the pars marginalis, the bifid nature of the internal end of the posterior central sulcus contouring the pars marginalis, and the lesser thickness of the posterior central gyrus compared with the precentral gyrus. The indirect method is less precise and is used when the direct method is unsuccessful. The central sulcus is identified on the sagittal images and, using the lateral view of the skull as a reference image, the topographic information is transferred to the axial images. PMID- 9417479 TI - Reviewer of the year: an excellent vintner. PMID- 9417478 TI - [An experimental study of the treatment of aneurysms using an intravascular prosthesis]. AB - Intracranial aneurysms occur frequently with the risk of major damage. Neurosurgery or endovascular techniques can be used for treatment. Current techniques are not well adapted for aneurysms with a wide implantation (or neck). The aim of this experimental work was to study a technique for treating aneurysms which can be used for wide neck aneurysms. A metal stent is implanted facing the neck of the aneurysm to allow occlusion. In the first part of the study, the stent was inserted alone. Ten aneurysms were created surgically in five dogs. The stents were positioned facing seven of the ten aneurysms. The stent led to immediate occlusion of the aneurysm in six of the cases. One aneurysm remained patent despite the correct position of the stent. One dog developed secondary thrombosis of the carotid. Three dogs have been followed for sixty days after insertion of the stent. Two aneurysms thrombosed and one was patent. Since these results were less than satisfactory, a second part of the study was undertaken to cover the stent with a fragment of the autologous vein. Results in five aneurysms, evaluated three and eight weeks after treatment, showed partial or total repermeabilization of the aneurysms. In the third part of the study, we associated stents and detachable coils. Twelve aneurysms of the carotid artery in pigs were thrombosed and two aneurysms were completely occluded after stent implantation. In the nine other cases, the aneurysms were completely occluded after stent implantation. In the nine other cases, the aneurysms remained patent despite the stent and treatment was then completed with a coil. Six of the animals have been followed for thirteen weeks. One carotid artery thrombosed. In the five other cases, the carotid arteries were patent and the aneurysms were occluded at the control angiogram. Histology results showed the presence of a fibromuscular endothelialized neointima at the neck of the aneurysm in four of the five cases. The technique described here could be proposed for the treatment of wide-neck aneurysms implanted low on the carotid artery in man. Improvements should render the system more flexible. PMID- 9417480 TI - The effects of social context and defensiveness on the physiological responses of repressive copers. AB - In previous research (T.L. Newton & R.J. Contrada, 1992), social context was found to moderate exaggerated physiological reactivity among individuals identified as using a repressive coping style. In this experiment, 119 undergraduates were classified into low-anxious, high-anxious, repressor, and defensive high-anxious coping categories. All participants completed a stressful speech task under either a public or private social context condition. The experimental social context was related to physiological reactivity and self reported affect but did not moderate reactivity among repressive copers. Additionally, reactivity among repressive copers was not attributable to high defensiveness alone. Consistent with a theory of emotional inhibition, nonspecific skin conductance responses, but not heart rate, discriminated between repressors and nonrepressors. PMID- 9417481 TI - Signalling an end to transplant rejection? PMID- 9417482 TI - The ethics industry. PMID- 9417483 TI - Calculating coauthors' contributions. PMID- 9417484 TI - [The 3rd Dresdner Selenium Symposium. 24-25 May 1997]. PMID- 9417485 TI - [Does selenium prevent peroxynitrite formation from NO in vascular surgery interventions? A clinical study]. AB - Nitric oxide seems to play a protective role in the pathogenesis of arteriosclerosis. It appears to protect the tissue during reperfusion in vascular surgery. High values of NO show an inverse correlation to the severity of arteriosclerosis. In our opinion selenium should be substituted in order to enhance the activity of the glutathione peroxidase. This means an effective protection against the high production of peroxinitrite in patients with arteriosclerosis. A comparison of the corresponding redox pairs SeO3(2-)/SeO4(2-) and HOONO/HONO allows the conclusion that sodium selenite can be a direct antioxidant thereby. PMID- 9417486 TI - [Selenium administration in sepsis patients]. AB - BACKGROUND: It has been hypothesized that low serum selenium concentrations, associated with low glutathione peroxidase activities in critical ill patients may contribute to decreased cleavage from free radicals and deteriorate the clinical outcome. PATIENTS AND METHODS: We therefore performed a controlled, prospective study including 42 patients with inflammatory response syndrome and an APACHE-II score > or = 15. Whereas the controls (Se-, n = 21) received 35 micrograms sodium selenite during the whole treatment period the selenium substitution group (Se+, n = 21) received additional 500 micrograms, 250 micrograms and 125 micrograms sodium selenite, each amount for 3 days. Clinical outcome was monitored by APACHE-III score, documentation of acute renal failure, respiratory insufficiency and the mortality rate until discharge from the hospital. RESULTS: The mean APACHE-II(III) score on admission was 20.6 (68.3) in the Se- versus 20.1 (61.0) in the Se+ group. Age, sex, underlying diseases, the serum selenium levels and glutathione peroxidase activities on admission were equally distributed in both groups. Selenium substitution was followed by a significant increase in serum selenium levels and glutathione peroxidase activity to normal levels, whereas in controls both parameters remained low. The APACHE III score significantly improved on day 7 (p = 0.018) and 14 (p = 0.041) in the Se+ group. Hemodialysis because of acute renal failure was necessary in 9 (Se-) versus 3 (Se +) patients (p < 0.04). Overall mortality rate in the Se+ group was 33.5% versus 55% in the Se- group (p = 0.13). A subanalysis of those patients with an APACHE-II score > 20 (n = 10) in each group revealed a significant reduction in mortality from 70% to 30% (p = 0.013). No negative side effects of selenium were seen. CONCLUSION: Selenium substitution significantly improves clinical outcome and reduces the incidence of acute renal failure. PMID- 9417487 TI - [Effect of selenium administration on various laboratory parameters of patients at risk for sepsis syndrome]. AB - BACKGROUND: Low selenium plasma levels were often measured in ICU patients with polytrauma, major surgery or various severe diseases. Activation of selenium dependent functions of the antioxidant metabolism and the immune system is suggested to be causally. METHODS: In a prospective randomized clinical trial including 24 critically ill patients we investigated the plasma levels of selenium, malondialdehyde, glutathione, elastase, fT3, fT4, TSH, IL-2R, IL-6 and IL-8 with and without parenteral selenium supplementation for 3 weeks (study design: week 1: twice 500 micrograms daily, week 2: once 500 micrograms, week 3: three times 100 micrograms sodium selenite). RESULTS: Following 24 hours of supplementation selenium plasma levels were normalized. Malondialdehyde level decreased in the therapy group significantly beginning at day 3. In the control group we observed increased malondialdehyde values, a disturbed glutathione metabolism and an elevated elastase activity. fT3-values were diminished at day 0 in all patients. In the therapy group we measured a gradual fT3 restoration. In the control group a reactive TSH increase was observed. Selenium supplementation did not lead to an excessive stimulation of IL-2R, IL-6 or IL-8. CONCLUSIONS: 1. Rapid normalization of selenium plasma levels can be achieved with the applied selenium dosage. 2. Parameters of radical metabolism are significantly reduced following selenium administration. 3. T3 synthesis correlates closely with the selenium levels. 4. Excessive stimulation of the immune system does not appear in the applied dosage. PMID- 9417488 TI - [Selenium administration in severe inflammatory surgical diseases and burns in childhood]. AB - PATIENTS AND METHOD: Substitution of selenium was performed in the University Clinic of Paediatric Surgery in Dresden in the time from 1994 to 1996 in 34 children aged 1 to 16 years with severe inflammatory surgical diseases as well a s widespread burns. Seven further patients have been examined within this time who have not received substitution of selenium as preliminary comparison group. All these patients fulfilled the criteria of "Systemic Inflammatory Response Syndrome" (SIRS). The following paraclinical parameters were examined: white cell count, interleukin 6, C-reactive protein, fibrinogen, malondialdehyde, activity of glutathione peroxidase in plasma and level of selenium in plasma and whole blood. RESULTS: Patients with initially low level of selenium who received substitution of selenium reached normal ranges more quickly than patients without substitution. Originally partly elevated values of malondialdehyde as sign of increased peroxidation of lipids were normalized under substitution of selenium. Initially low activity of selenium level in plasma showed a clear increase under substitution of selenium as sign of increased protection of the cell membrane. CONCLUSION: The substitution of selenium in children with SIRS is a supportive therapy. PMID- 9417489 TI - [Importance and physiologic nutritional requirement of the selenium trace element within the scope of parenteral nutrition (TPN)]. AB - BASIS: Selenium-responsive clinical manifestations of selenium deficiency and elucidation of the biochemical and molecular biological basis of the essentiality of selenium give evidence for the biological importance of the trace element in human nutrition. CONCLUSION: The dietary parenteral selenium requirement can be calculated on the basis of the maximal gene expression of the selenoprotein plasma glutathione peroxidase (plGPx). In total parenteral nutrition a daily requirement of 0.01 mumol/kg body weight for adults and 0.025 mumol/kg body weight for children can be seen as adequate and safe. PMID- 9417490 TI - [Effect of selenium administration on various laboratory parameters in patients with acute pancreatitis]. AB - BACKGROUND: Recent studies presented evidence that activation of oxygen derived free radicals occurs in patients with acute pancreatitis. The purpose of this study was to evaluate the effect of sodium selenite as a possible antioxidant therapy in acute pancreatitis. PATIENTS AND METHOD: 16 patients with moderate form of acute pancreatitis received a high dose of sodium selenite. Selenium in serum and whole blood, zinc, copper, manganese, superoxid dismutase (SOD), glutathione peroxidase (Gpx) and malondialdehyde (MDA) were determined (before selenium substitution, 3 days later, 8 days later, before demission). No selenium deficiency could be detected before selenium substitution. RESULT: The selenium therapy caused a significant increase in selenium, a moderate increase in activity of Gpx, a significant decrease in activity of MDA, whereas SOD remained unchanged. CONCLUSION: Concerning the particular point of view of "deficiency management", there is no need of selenium substitution in patients with a moderate form of acute pancreatitis in our region. The highly normal selenium concentration we established by our therapy is possibly connected with a decrease of the oxidative stress in acute pancreatitis. More clinical follow-up studies with more patients, who have different grades of severity of the acute pancreatitis, and besides that a control group of patients without selenium substitution, are necessary for evaluating the clinical relevance of our results. PMID- 9417491 TI - [Selenoproteins in bone, gastrointestinal tract and thyroid gland of the human]. AB - BASIS: Selenium is an essential trace element, which is incorporated as selenocysteine (secys) into specific proteins in a regulated fashion. In the presence of a hairpin loop structure within the 3' untranslated region of the mRNA the opal stop codon UGA is coding for selenocysteine. Selenoprotein functions are dependent on secys incorporation. Members of the family of deiodinases as well as the family of glutathione peroxidases, selenoprotein P and thioredoxin reductase are selenoproteins. DISCUSSION: Bone, the intestine and the thyroid rely on antioxidant systems against potential cell and DNA damage through endogenous and environmental peroxides and reactive oxygen species (ROS) potentially promoting inflammation and tumorigenesis. Optimized cell defense through antioxidant selenoproteins requires optimal selenium supplementation of the organism. We have analyzed the expression of selenoproteins in these tissues, thus providing molecular tools to further elucidate optimal selenium supply on a cellular level. CONCLUSION: Clinical intervention studies that focus on the development of disease must confirm the relevance of optimized selenium supply for the pathogenesis, prevention and therapy of metabolic bone disease as well as chronic (autoimmune) inflammation and tumorigenesis in the thyroid and intestine. PMID- 9417492 TI - [Selenium substitution in acute myocardial infarct]. AB - BACKGROUND: Previous examinations have demonstrated decreased selenium levels in serum and full blood in patients with myocardial infarction. PATIENTS AND METHOD: 28 patients received a selenium treatment additional to the usual treatment of myocardial infarction. 19 patients with myocardial infarction with no supplementary selenium treatment served as a control group. Selenium levels in serum, full blood and urine were measured and the complications of the myocardial infarction documanted. RESULTS: There was a significant increase of serum and full blood selenium and glutathione peroxidase levels under i.v. selenium therapy in the acute phase of myocardial infarction (first to third day). Left heart failure more rarely occurred in the selenium group (20%) than in control patients (57%). Acute tachycardial cardiac rhythm disturbances such as ventricular extrasystoles and couplets diminished in both groups; ventricular extrasystoles decreased in the selenium group. CONCLUSIONS: Selen should be substituted in patients with acute myocardial infarction and decreased selen levels. It would be useful to carry out a prospective double-blind study. PMID- 9417493 TI - [Selenium concentration in erythrocytes of patients with rheumatoid arthritis. Clinical and laboratory chemistry infection markers during administration of selenium]. AB - PATIENTS AND METHODS: Seventy patients with definitive rheumatoid arthritis were matched to built 2 groups, which were double-blind and randomized allocated to supplementation with sodium-selenit 200 micrograms/d or placebo for 3 months, each. Both groups were given fish oil fatty acids (30 mg/kg body weight), DMARDS were continued throughout the study, while variations in steroids or NSAD were admitted. RESULTS: Selenium concentrations in erythrocytes of patients with rheumatoid arthritis were 85.1 +/- 26 micrograms/l, and significantly lower than found in an average German population (123 +/- 23 micrograms/l). During the observation period of 3 months normal selenium concentrations were not restored, despite supplementation higher than RDA. At the end of the experimental period the selenium supplemented group showed less tender or swollen joints, and morning stiffness. Selen-supplemented patients needed less cortisone and NSAD than controls. In accordance with clinical improvement we found a decrease of laboratory indicators of inflammation (C-reactive protein, alpha 2-globuline, prostaglandin E2). CONCLUSION: No side effects of supplementation with selenium were noted, which can be considered as adjuvant therapy in patients with rheumatoid arthritis. PMID- 9417494 TI - [Selenium administration in patients with sepsis syndrome. A prospective randomized study]. AB - PATIENTS AND METHOD: In this study the effect of antioxidative therapy with sodium selenite was investigated in patients with systemic inflammatory response syndrome (S. I. R. S.) and multiple organ failure. 40 patients were included in this prospective randomized study. The patients were observed over a period of 28 days. The letality rate within 28 days was excepted as main criteria. The Apache II and the MOF-Score of Goris were used as clinical parameters. 20 patients were treated with sodium selenite over a period of 28 days. RESULT: This antioxidative therapy reduced the letality rate from 40 to 15%. PMID- 9417495 TI - [Selenium in phenylketonuria patients. Effects of sodium selenite administration]. AB - PATIENTS AND METHOD: 17 patients (8 female, 9 male; age 8.2 +/- 3.7 years) with phenylketonuria under phenylalanin restricted diet were investigated prior to and after 3 months of selenium substitution (sodium selenite, 115 micrograms Se/m2 BSA/d). Different parameters in blood were determined: selenium, glutathione peroxidase (Gpx) activity, thyroid hormones, blood cell count, lymphocytic antigen expression, muscle function and -enzymes, cardiac ultrasound. RESULTS: The main significant results of selenium substitution are: increased plasma selenium, blood cell selenium, plasma-Gpx activity and left ventricular cardiac index as well as decreased plasma thyroxin, free thyroxin, reverse triiodthyronin, total cholesterol, mean erythrocyte and thrombocyte volume and lymphocytic CD2 expression. CONCLUSION: The data indicate metabolic and functional signs of selenium deficiency in patients with phenylketonuria without selenium substitution. We conclude that, despite of lacking clinical symptoms, a selenium supply in phenylketonuria patients under diet is necessary and should be performed with usefull peroral sodium selenite (115 micrograms Se/m2 BSA/d) initially, followed by a dosage between 30 and 60 micrograms Se/m2 BSA/d). PMID- 9417496 TI - [Levels of antioxidants after cesarean section and administration of Multibionta N, Inzolen and selenase]. AB - PATIENTS AND METHOD: In 29 women with the necessity to terminate pregnancy via Cesarean section, lipid peroxidation and antioxidative state were investigated before and 24 hours after the surgical intervention as well as after substitution of antioxidants and trace elements. RESULTS: The results indicate that administration of antioxidants protects at least partially from consequences of surgically induced oxidative burden. PMID- 9417497 TI - [Sex hormones and antioxidant status]. AB - Physiologically, an influence of sex steroids on the antioxidative capacity can be seen at least as a short-term effect. First of all the steroid effects are due to the estrogen component rather than the progestin component. The antioxidative potential of natural estrogens is several times higher than that of vitamin E. Long-term studies should be performed to verify this in perimenopausal hormone replacement. PMID- 9417498 TI - [Behavior of thiobarbituric acid reactive substances, alpha-tocopherol, glutathione and selenium during hypertension in pregnancy]. AB - PATIENTS AND METHOD: In a longitudinal study the changes of serum or plasma levels of TBARS, alpha-tocopherol, glutathione and selenium were investigated comparing 46 pregnant women with hypertensive syndrome and 18 women with normal pregnancies. RESULTS AND CONCLUSIONS: A significant increase in TBARS serum level was found in women suffering from hypertension. There was no correlation between the severity of hypertension and the extent of the TBARS rise. An absolute deficiency of the antioxidants alpha-tocopherol, glutathione and the elements selenium, calcium, magnesium, iron and zinc as a cause or a consequence of hypertension in pregnancy could be excluded. Low serum levels of copper and selenium could decrease the activity of glutathione peroxidase (Gpx) resulting in higher levels of TBARS and glutathione during the last trimester of pregnancy and delivery. Increased concentrations of iron in women with hypertensive syndrome may affect the formation of lipid peroxides. The outcome of children was unaffected by increased lipid peroxide levels when an antihypertensive therapy was consequently performed. PMID- 9417499 TI - [Reference values for blood and serum selenium in the Dresden area]. AB - BACKGROUND: To ensure a correct interpretation of patient's data the regional differences in the supply with selenium have to be taken into consideration. PATIENTS AND RESULTS: In 256 healthy women and men from the area of Dresden aged 20 to 62 years the selenium reference values were examined in blood serum by 1.09 +/- 0.17 (0.75 to 1.43) and in whole blood by 1.29 +/- 0.21 (0.87 to 1.71) mumol/l. There was no dependence upon age and sex and no influence of alcohol, tobacco and vegetarian diet was found. Consumption of beer yeast and frequent fish meals caused improvement of the selenium status. CONCLUSION: In area of Dresden, similar to the whole of Germany, a marginal selenium supply exists. Therefore it is of high importance to consider a balanced nutrition and to control the selenium status especially in serious acute diseases and in intensive care. PMID- 9417500 TI - [Blood selenium content after conditioning and during the course of bone marrow transplantation in children with malignant diseases]. AB - PATIENTS AND RESULTS: Prior to bone marrow transplantation (BMT), at the end of the conditioning phase, we found in 42 investigated children with malignant diseases subnormal lowered plasma- and blood selenium levels. Parallel to the diminished selenium status the plasma glutathione peroxidase activity (Gpx) was not reduced as it is in selenium deficiency, but markedly elevated and probably reflecting cytolytic processes. In the group of combined conditioning (fractionated total body irradiation plus chemotherapy) we found significantly more elevated plasma Gpx values in comparison to the only-chemotherapy group. The renal selenium excretion was elevated during the whole observation and could be caused by disturbed tubular function. CONCLUSION: We conclude, that in the situation of BMT a selenium substitution in a dosage of at least 1 to 2 micrograms Se/kg/d is necessary. Patients' selenium status should be monitored by analyses of plasma- and blood selenium contents. PMID- 9417501 TI - [Role of sodium selenite as an adjuvant in radiotherapy of rectal carcinoma]. AB - BACKGROUND: In various epidemiologic studies an association of low selenium blood levels and reduced glutathione peroxidase with an increased risk of cancer incidence was described. The antitumoral therapy and a suboptimal nutrition could intensify this deficiency. Every reduction of disease related and therapeutic caused symptoms may improve life quality. APPLICATION: We report our preliminary experiences in the adjuvant radiochemotherapy of advanced rectal cancer (UICC II/III) corresponding to the NCl recommendation. An oral selenium supplementation was carried out with 2000 micrograms Na2SeO3 after every course of fluorouracil chemotherapy and daily 400 micrograms Na2SeO3 after irradiation of tumor region and lymph nodes. A weekly life quality assessment was explored with special interest in diarrhea, dysurie, pain, appetite, nausea and emesis. CONCLUSION: Damages to normal tissue specially to DNA enzymes and membranes caused by free radicals is one mechanism in tumorgenesis, tumor progression and therapeutic consequence. A radioprotective effect of selenium is verified by in vitro and in vivo data. Our data show that oral selenium intake in rectal cancer patients is easily tolerated with no side effects. Improving life quality and secondary cancer prevention with supplementation of selenium has to be proven in prospective randomized studies. PMID- 9417502 TI - [Selenium in peroxide metabolism]. AB - Out of the growing number of so far known mammalian selenoproteins four are peroxidases. Their common catalytic mechanism involves redox shuttling of a selenocysteine residue in the active site, where it forms a characteristic catalytic triad with hydrogen-bonded tryptophan and glutamine residues. These peroxidases differ in tissue distribution, substrate specificity, regulation, responsiveness to selenium restriction, and likely in their biological role. Cytosolic glutathione peroxidase, which predominates in balancing hydroperoxide toxicity, rapidly declines in selenium deficiency. Prophylactic selenium supplementation is considered reasonable also in asymptomatic moderate selenium deficiency whenever a clinical condition known to cause oxidative stress has to be anticipated. PMID- 9417503 TI - [Administration of selenium in lymphedema]. AB - AIM: To investigate the effectiveness and dosage of selenium in the treatment of lymphedema. RESULTS: Intensive treatment with decongestive physical therapy in combination with sodium selenite shows significant improvement in the reduction of the volume of the limb and in the histophysiology of secondary lymphedema after mastectomy or Wertheim-operation. The incidence of skin infections decreased significantly. CONCLUSION: The anti-inflammatory effect should be considered in the treatment of lymphedema. PMID- 9417505 TI - [30th Scientific meeting of the German-speaking Association of Mycology. Kiel, 19 22 September 1996]. PMID- 9417504 TI - [Selenium, antioxidant status and radical/reactive oxygen species in medicine]. AB - BACKGROUND: In all organisms with a metabolism of oxygen, radicals/reactive oxygen species appear. Normally a balance exists between formation and destruction of radicals. Many diseases show a disturbance of this balance with the destruction of biological structures. Results in patients with sepsis, terminal renal failure, bone marrow transplantation and cancer are given. CONCLUSION: To avoid a possible pro-oxidative effect (vitamins C and E) and intoxications (selenium), before substitution a monitoring is necessary. PMID- 9417506 TI - [Aspergillus fumigatus and Chaetomium homopilatum in a leukemic patient. Pathogenic significance of Chaetomium species]. AB - From the tracheal secretion of a leukaemic patient Aspergillus fumigatus and Chaetomium homopilatum was isolated. Radiographically (HR-CT) an invasive pulmonary mycosis was diagnosed from which the patient died. As an autopsy was not performed, the role of the isolated fungi could not be clarified safely. Aspergillus fumigatus is supposed to have been responsible for the invasive mycosis. The etiopathological significance of Ch. homopilatum remained unclear. The isolation of Ch. homopilatum was a reason for reviewing the genus Chaetomium. In the literature 18 reported cases of infections by Chaetomium sp. were found. Ch. globosum was the most prevalent species and caused mostly onychomycosis. Ch. strumarium and Ch. atrobrunneum caused brain infections. The predisposing factor in case of onychomycosis and cutaneous lesions was a trauma, and the systemic mycoses were a consequence of leukaemia, renal transplantation, intravenous drug use or renal failure. The reported cases show, that Chaetomium sp. may cause infections, if predisposing factors are present. Therefore the isolation of Chaetomium sp. in clinical specimen should not regarded as a contamination, and the possible etiopathological significance should be clarified. PMID- 9417507 TI - [Hormones, fungi and skin]. AB - It is indicated by some epidemiological and clinical observations, that steroidal hormones may belong to those factors that are capable to influence the clinical courses of mycotic infections in man. Several fungal species, including pathogenic ones, are able to produce or metabolize steroidal hormones, or their growth can be affected by such hormones. Since, on the other hand, the steroid responsive human skin is also capable to synthesize and convert steroidal hormones, the relationship between pathogenic fungi and host may be influenced by hormonal mediators in dermatomycoses. PMID- 9417508 TI - [Mass development of Stachybotrys chartarum on compostable plant pots made from recycled paper]. AB - After handling plants grown in decomposable pots made of recycling paper, three women working in a big horticulture developed very painful inflammated efflorescences at the finger-tips, followed by scaling off the skin. The pots appeared to be very mouldy. Black masses of conidia of Stachybotrys chartarum and perithecia of Chaetomium globosum were identified on almost every pot. Apart from various other fungal genera, Trichoderma und Acremonium were frequently detected. Considering the observed symptoms, special attention was payed to the mycotoxin producing species St. chartarum. To evaluate the inhalative spore load, air sampling was performed. The detection of St. chartarum in the air was only possible with the spore trap (sampling of particles) but not with the Andersen sampler (detection of colony forming units). Without moving the pots, measurements yielded values of 30-100 St. chartarum conidia per m3 of air. The concentration of air-borne conidia increased drastically by handling the pots, thus attaining up to 7,500 conidia per m3 of air for St. chartarum only. The occurrence of St. chartarum in such amounts is alarming because of possible toxin production. In addition, the allergenic stress by fungal spores has to be emphasized. The results are discussed with regard to general medical-mycological aspects related to the degradation of environmentally-friendly decomposable materials. PMID- 9417509 TI - [Dermatophytes do not produce sialidase in vitro]. AB - Sialidase (EC 3.2.1.18) is a pathogenicity factor of many microorganisms, and may also play a role in adhesion of dermatophytes to the epithelia of their hosts by the hydrolytical cleavage of terminal, negatively charged sialic acids of glycoconjugates on the cell surfaces, thus allowing fungal lectins to bind to the subterminal sugars. Therefore, 116 strains of seven species of dermatophytes were investigated for sialidase production. Two highly sensitive, quantitative sialidase assays were applied to cell homogenates and culture supernatants from seven different media of the fungi, but were always negative for sialidase activity. However, sialidase activity was always detected in Ophiostoma stenoceras used as a positive control cultivated in parallel; the enzyme was inducible by sialylated mucins. A sialidase-dependent pathomechanism for dermatophytes appears unlikely based on the results presented. PMID- 9417510 TI - [Mycoses in patients with psoriasis or atopic dermatitis]. AB - Both, psoriasis and atopic dermatitis are multifactorial diseases with an unknown pathogenesis. To elucidate the influence of fungal infections in the onset or recurrence of those inflammatory skin disease we determined the frequencies of Candida and dermatophyte infections of the skin and orointestinal tract concomitantly seen in patients with psoriasis or atopic dermatitis. We analyzed the files of more than 40,000 in-patients of the Department of Dermatology, Kiel. Sex- and age-adjusted relative risk were calculated. The results demonstrate that patients with psoriasis (N = 3006) presented with a decreased rate of tinea. This was significant for tinea corporis (RR = 0.13, p < 0.01). Candida infections of the skin were seen more often in psoriasis patients compared to controls. Differentiating between Type I (early onset) and Type II (late onset) psoriasis only Type I psoriasis patients presented with decreased dermatophyte infections and increased Candida colonization of the intestinum. However, patients with Type II psoriasis demonstrated an increased rate of candidosis cutis and candidosis oris as compared to controls. Patients with atopic dermatitis (N = 1808) displayed a decreased overall incidence of tinea and Candida infections. Furthermore, in patients with atopic dermatitis a Candida colonization of the orointestinal tract was found elevated (RR = 1.51, p < 0.01), whereas tinea corporis (RR = 0.24, p < 0.01) and candidosis cutis (RR = 0.30, p < 0.001) was found decreased compared to controls. Our results show that the influence of fungal infections on the two skin diseases investigated is not as strong as often considered. The increased relative risk in patients with atopic dermatitis to present with Candida colonization in the digestive tract or vice versa may contribute to the pathogenesis of atopic dermatitis. PMID- 9417511 TI - [High-dose therapy with fluconazole > or = 800 mg/day. Review]. AB - The clinical use of fluconazole in dosages > or = 800 mg/day has been reported in about 900 patients against candidemia, oropharyngeal candidiasis and cryptococcal meningitis in HIV-infected patients as well as for initial therapy of endemic mycoses. Especially in patients with life-threatening infections caused by Candida spp., Cryptococcus neoformans and Coccidioides immitis, the results of a limited number of dose-finding trials with non-neutropenic and HIV-infected patients show dose-dependent response rates. These findings strongly advocate the application of high dose-fluconazole; their evaluation, however, still awaits final clarification. The good safety profile for dosages up to 2000 mg/day and the linear, predictable pharmacokinetics up to 1600 mg/day indicate the excellent tolerability of fluconazole in the clinical situation which justifies prospective, randomized clinical trials with treatment groups as homogeneous as possible for further evaluation of the optimum dosage and duration of treatment. PMID- 9417512 TI - [Effect of keratin on the efficacy of fluconazole]. AB - In this study we have investigated the influence of keratin on the efficacy and pharmacokinetics of fluconazole and additional azole antifungal agents. It is well known that the penetration and distribution of oral antifungals is strongly influenced by plasma protein binding. Especially, itraconazole and ketoconazole show a high binding affinity to plasma proteins, whereas fluconazole binds only with 12%. All of these antifungals, however, are accumulated in the stratum corneum of the skin. These observations have stimulated interest, if structure proteins of the skin like keratin interact with antifungals may explain of the accumulation process. Therefore we have measured the binding kinetics between keratin and the azoles. Keratin from sheep wool was degreased, purified and incubated with azole antifungals. After defined incubation periods the azoles were extracted and assayed by thin layer chromatography following UV-detection. There was a specific binding between keratin and all of the used substances. Interestingly, the binding affinity of fluconazole to keratin was much higher than to plasma proteins. Thus our observations indicate that the accumulation in the stratum corneum is a consequence of the interaction between keratin and azole derivatives. PMID- 9417513 TI - [Evaluation of a breakpoint test for determination of fluconazole susceptibility of yeasts]. AB - Using the breakpoint test at 1 g/ml and 4 g/ml fluconazole, a minimal inhibitory concentration (MIC) of < or = 4 g/ml fluconazole was determined against 78.5% of the 1254 clinical yeast isolates. When compared with the micro broth dilution test, none of a subset of 128/1254 strains had a higher MIC in the dilution test than in the breakpoint test, however, in 43.0% of the 128 strains the MIC was lower in the micro broth dilution test when compared to the MIC of the breakpoint test. In a subset of 94 strains with an MIC of > 4 g/ml fluconazole determined in the breakpoint test, the elevated MIC could be confirmed only in 45.7% of the strains when using the micro broth dilution test. The percentage of breakpoint test confirmation as well as the number of strains with decreased susceptibility towards fluconazole (> 4 g/ml) were species dependent, thus, the number of decreased-susceptible Candida albicans strains was smaller than that of C. glabrata or other Candida species such as C. krusei, C. inconspicua and some C. tropicalis strains. The breakpoint test allows to identify susceptible strains with a high accuracy. Strains with an MIC > 4 g/ml fluconazole should be tested in the micro dilution test to confirm decreased susceptibility and thus to indicate the need for higher dosage of fluconazole or a change of the antifungal therapy. The breakpoint test proved to be a rapid and reliable screening test. PMID- 9417514 TI - [Use of fluconazole as antimycotic prophylaxis in radiotherapy of patients with head and neck tumors]. AB - The aim of the present study was to investigate the incidence of Candida stomatitis and resulting interruptions in radiation therapy in 50 patients suffering from squamous cell carcinomas of head and neck region receiving fluconazole (100 mg/d) in comparison to a historical control group without specific prophylaxis. 20 of the control patients (40%) demonstrated Candida stomatitis with 7 of them (14%) requiring interruptions of radiation therapy. In contrast, none of the patients with fluconazole had evidence of Candida stomatitis and subsequent interruption of anticancer therapy. Laboratory monitoring for the presence of Candida species was performed in 15 other patients before and after therapy with fluconazole. Candida albicans was identified less frequently after therapy when compared to the pretreatment status. However, C. glabrata and C. krusei were isolated in some of the patients probably due to decreased drug susceptibility of these species. The results demonstrate the clinical usefulness of prophylactic fluconazole applications in patients suffering from head and neck tumors with the aim to reduce Candida stomatitis and resulting interruptions in radiation therapy. PMID- 9417515 TI - [Fluconazole-resistant Candida species from HIV infected patients with recurrent Candida stomatitis: cross resistance to itraconazole and ketoconazole]. AB - In vitro susceptibility to fluconazole of Candida species isolated from 83 HIV infected patients treated with fluconazole because of recurrent Candida stomatitis was monitored over a period of two years. A microdilution assay with high-resolution antifungal assay (HR) medium and RPMI 1640-medium were compared. In vitro less susceptible and fluconazole resistant C. species were observed in 23 patient at the end of the study. The Candida isolates recovered from oral rinsing fluids at the beginning and the end of study were tested for crossresistance to itraconazole and ketoconazole. Susceptibility to ketoconazole and to itraconazole was reduced using RPMI 1640-medium. Susceptibility of the isolates to fluconazole was not influenced by the assay medium. In 21 patients in vitro fluconazole resistant or less susceptible C. albicans were observed. 9 of 21 isolates showed crossresistance to itroconazole and ketoconazole, in 10 isolates only crossresistance to itraconazole was observed. During fluconazole treatment double infections due to combination of C. albicans and C. glabrata or C. krusei increased from 20% to 78% C. krusei was resistant to the three azoles. C. glabrata was less susceptible (4-8 mg/l) or resistant (> 8 mg/l) to fluconazole and resistant to itraconazole and ketoconazole High dosed intravenous fluconazole treatment with 400 to 600 mg daily failed in 11 patients with fluconazole resistant C. albicans and in 3 (3/10) patients with les susceptible C. albicans isolates. PMID- 9417516 TI - [Involvement of secretory Candida proteinases in the adherence of C. tropicalis blastoconidia in a cell culture model]. AB - The influence of the heterologous acid secretory Candida proteinases on the adherence of the non-proteinase secreting strain of C. tropicalis DSM 4959 to epitheloid cells (vero line) was examined. The proteinases of the following Candida strains were used: C. albicans ATCC 10261 (serotype A), C. albicans ATCC 48867 (serotype B), C. tropicalis DSM 4238. The assays were performed with the previously described in-vitro-adherence test [1] using the following principle steps: Candida proteinases and C. tropicalis blastoconidia were incubated with verocells in microtest plates in phosphate-buffer in the range of pH 4.0 to pH 7.0. Adherent Candida cells were detected according to Filler et al. [2] with anti-Candida-mannoprotein antibodies and a secondary anti-rabbit-peroxidase conjugate. Compared to controls with denaturated proteinases, the photometric evaluation of adherent C. tropicalis cells showed, under optimal conditions, an augmentation of the adherence due to the Candida proteinases of about 50%. The optimum of this adherence augmentation was in the range of pH 5.5 which is outside the general activity optimum of Candida proteinases (pH 3). The degree of purity of these proteinases had no marked influence on the adherence. The specificity of the proteinase dependent adherence augmentation could be demonstrated with the enzyme inhibitor Pepstatin A. C. tropicalis blastoconidia supplemented by pepstatin A and active Candida proteinase adhered in the same range as with denaturated proteinases in control tests. Our results suggest a function of Candida proteinases in the adherence process of blastoconidia to epithelia. PMID- 9417517 TI - [Tinea follicularis presenting as trichophytic Majocchi granuloma]. AB - We report on a 75 year old patient with a bronchial asthma treated at least for 15 years with low dose prednisolone. Under this treatment he developed a tinea follicularis and was demonstrated in our clinic with papulopustular skin lesions on both forearms, left malleolus and left thigh. We saw tender to touch granulomata on erythematosquamous atrophic skin. A dermatomycosis was diagnosed by isolation and identification of Trichophyton rubrum. In addition the onychomycosis of all finger- and footnails was caused by T. rubrum. Clinical and accessory clinical findings (histology) agreed with a 1883 described disease, granuloma trichophyticum Majocchi. An internal treatment with terbinafine 250 mg/d and topical with tioconazole cream completely cured the skin lesions and the nails. PMID- 9417518 TI - [Persistence and variability of yeasts isolated from hospitalized patients: a comparison of results from Rostock and Dresden]. AB - We investigated the yeast colonizations of hospitalized patients at time of the admission to hospital (< or = 3d; 1161 patients) and during stay in hospital (> 3d-several months; 568 patients). At admission to hospital 58% of patients had yeasts in one of the investigated specimens. During stay in hospital the part of patients with yeasts increased up to 81.7%. We established remarkable differences in proof of yeasts in patients of different area of risk. The spectrum of yeasts of the patients in Rostock and Dresden shows a similar shift in frequency of the different Candida species. C. albicans was the predominant yeast. But during hospitalization we saw an elevation of patients with C. glabrata infection from 7.4 to 22.5% and C. krusei infection from 2.8% to 11.8%. There were a remarkable correlation to the area of risk. In 30.8% of the patients we observed a change in yeast spectrum: from negative cultures to positive specimens or from one Candida species to another one. PMID- 9417519 TI - [Patterns of Candida esophagitis in cancer and AIDS patients: histopathological study of 23 patients]. AB - Candida oesophagitis is a common concomitant disease in neutropenic cancer patients after chemotherapie as well as in HIV-patients. In order to characterize the features of oesophagitis in each population, we reviewed the medical history and pathology records of 23 patients (18 cancer-patients, 5 HIV-patients) with culture and autopsy-proven Candida oesophagitis. Histopathological patterns of morphology, invasion, angioinvasion and inflammation were evaluated. Virtually all patients, 17/18 cancer- and 5/5 HIV-patients, had a history of previous mucosal candidosis or candidemia. There was a significant difference histopathologically in depth of invasion of the Candida-organisms between cancer and HIV-patients. Only in HIV-patients organisms were observed within the muscularis propria and the adventitia (2/5 vs 0/18; p = 0.04). The frequency of angioinvasion (12/18 vs 3/5) was similar in both groups. Neutropenia (< 500/microliter) was present in 12 (68%) of 18 cancer patients vs 0/5 HIV-patients (p = 0.01). Correspondingly there was a significant higher PMN/MN ratio in the oesophageal inflammatory infiltrate in HIV-patients, reflecting chemotherapy induced neutropenia in cancer patients (p = 0.02). Oesophageal candidosis in HIV patients may be highly invasive despite the presence of neutrophils. These findings suggest an impaired inflammatory response of HIV-patients to invasive candidosis, leading to impaired mucosal host defence. PMID- 9417520 TI - [Candida identification: experiences with the Vitek automated system]. AB - We report about our experiences obtained with the Vitek system (bioMerieux, Nurtingen) for identification of 1160 clinical Candida isolates. The Vitek system correctly identified 1005 (86.6%) Candida isolates (probability > or = 85%): 776 (66.9%) strains were identified after 24 h of incubation, 229 (19.7%) required 48 h of incubation. In 4.4% (n = 51) we found results with a lower probability (< 85%). After 24 h of incubation 38 (14.6%) of the 260 C. tropicalis isolates tested were misidentified as C. parapsilosis; we obtained the correct results after 48 h of incubation. 104 (9%) isolates were wrongly identified after 48 h of incubation. The majority of these strains (n = 82) were biochemically minimally reactive Candida species. Overall, the Yeast Biochemical Card system provides a reliable method for the rapid, automated identification of medically important Candida species. PMID- 9417521 TI - [Therapy of infections caused by dematiaceous fungi]. AB - Since dematiaceous fungi belong to the group of rare infectious agents causing mycoses, therapeutic recommendations are mainly deduced from observations of single cases. In cases of eumycetoma or focal phaeohyphomycoses of the central nervous system (e.g. caused by Cladophialophora bantiana or Exophiala dermatitidis) and in certain cases of chromoblastomycoses, the resection in toto is the therapy of choice, which may be accompanied by antimycotic medication. As antimycotic therapy ex juvantibus in case of phaeohy-phomycoses, a simultaneous application of itraconazole and 5-fluorocytosine is presently most promising. The success depends on an adequate duration of therapy. PMID- 9417522 TI - [Autopsy findings in patients with repeated demonstration of Candida glabrata]. AB - In our mycological laboratory we saw an increase of Candida glabrata isolates from 0.5% in 1993 to 22.2% in 1996, mainly isolates of high risk patients of intensive care units and oncological wards. We were interested to know whether this increase of C. glabrata is also corresponding to the number of endomycoses. Eighteen patients have been autopsied and examined histologically. During the hospital course we found repeatedly in all patients C. glabrata with or without C. albicans. Serological and culture results of this intensive care/oncological patients were indicative of an invasive endomycosis. In 12 of 18 patients there was no evidence of an invasive mycosis in autopsy tissues. Three patients had a typical aspergillosis of the lungs, which was not diagnosed during hospital course. In 5 patients, two of them with aspergillosis of the lungs, we found in kidney or lung tissue sporadic yeast cells, typified by size and missing pseudomycelium as C. glabrata. In no case a tissue reaction as an intravital defense reaction against these yeast cells was observed. Therefore, we interpreted the findings of C. glabrata in tissues as representing a transient fungemia and not as an endomycosis due to C. glabrata. PMID- 9417523 TI - Leukocyte reduction and ultraviolet B irradiation of platelets to prevent alloimmunization and refractoriness to platelet transfusions. AB - BACKGROUND: We conducted a multi-institutional, randomized, blinded trial to determine whether the use of platelets from which leukocytes had been removed by a filter or that had been treated with ultraviolet B irradiation would prevent the formation of antiplatelet alloantibodies and refractoriness to platelet transfusions. METHODS: Patients who were receiving induction chemotherapy for acute myeloid leukemia were randomly assigned to receive one of four types of platelets transfusions: unmodified, pooled platelet concentrates from random donors (control); filtered, pooled platelet concentrates from random donors (F PC); ultraviolet B-irradiated, pooled platelet concentrates from random donors (UVB-PC); or filtered platelets obtained by apheresis from single random donors (F-AP). All patients received transfusions of filtered, leukocyte-reduced red cells. RESULTS: Of 530 patients with no alloantibodies at base line, 13 percent of those in the control group produced lymphocytotoxic antibodies and their thrombocytopenia became refractory to platelet transfusions, as compared with 3 percent in the F-PC group, 5 percent in the UVB-PC group, and 4 percent in the F AP group (P< or =0.03 for each treated group as compared with the controls; there were no significant differences among the treated groups). Lymphocytotoxic antibodies were found in 45 percent of the controls, as compared with 17 to 21 percent in the treated groups (P<0.001 for each treated group as compared with the controls; there were no significant differences among the treated groups). Antibodies against platelet glycoproteins developed in 6 to 11 percent of the patients, with no significant differences among the four groups. CONCLUSIONS: Reduction of leukocytes by filtration and ultraviolet B irradiation of platelets are equally effective in preventing alloantibody-mediated refractoriness to platelets during chemotherapy for acute myeloid leukemia. Platelets obtained by apheresis from single random donors provided no additional benefit as compared with pooled platelet concentrates from random donors. PMID- 9417524 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 40-1997. A 23-year-old man with a mediastinal embryonal carcinoma and hematologic abnormalities. PMID- 9417525 TI - Probucol and multivitamins in the prevention of restenosis after coronary angioplasty. PMID- 9417526 TI - Probucol and multivitamins in the prevention of restenosis after coronary angioplasty. PMID- 9417527 TI - Probucol and multivitamins in the prevention of restenosis after coronary angioplasty. PMID- 9417528 TI - Myocardial ischemia with normal coronary arteries associated with thoracic myelitis. PMID- 9417529 TI - Intermittent etidronate and corticosteroid-induced osteoporosis. PMID- 9417530 TI - Management of hepatic encephalopathy. PMID- 9417531 TI - Spurious extreme reticulocytosis with an automated reticulocyte analyzer. PMID- 9417532 TI - An educational program to prevent disabling low back pain. PMID- 9417533 TI - An educational program to prevent disabling low back pain. PMID- 9417534 TI - [Prenatal diagnosis of triploidy and trisomy 21 through fetal erythroblasts isolated from maternal blood]. AB - BACKGROUND: A long-sought goal of medical genetics has been the development of prenatal diagnostic procedures that do not endanger the conceptus. The safety of noninvasive methods for prenatal diagnosis would be especially attractive because they could be extended to all pregnant women, regardless of their ages or histories. Noninvasive prenatal diagnosis for the entire population might be possible recovering fetal cells from maternal blood. For this purpose, we have studied fetal erythroblasts. MATERIALS AND METHODS: To evaluate the potential of the method for clinical use, we studied maternal blood samples from 11 women referred to us for prenatal diagnosis between 15 and 20 weeks of gestation. For simple and effective enrichment of fetal nucleated erythrocytes from peripheral maternal blood, we combined a triple density gradient and magnetic-activated cell sorting (MACS) of anti-CD71 transferrin receptor antibody labeled cells. The isolated cells were analysed by using dual-colour interphase fluorescent in situ hybridization (FISH) with X-, Y-, 18- and 21-specific DNA probes. RESULTS: Chromosomal abnormalities detected on enriched fetal cells include trisomy 21 and triploidy. CONCLUSIONS: Based on the current results it is suggested that the technique described here is a simple, fast, efficient and reliable method for non invasive prenatal diagnosis. PMID- 9417535 TI - ["Hyperferritinemia-cataract syndrome." Description of a new hereditary disease, from anamnesis to molecular diagnosis]. AB - A new autosomal-dominant genetic disorder, which has been recently identified by our group is described. The disease is clinically characterized by the combination of a substantial increase of serum ferritin and early-onset bilateral cataract. Moreover, it is clearly distinguishable from genetic hemochromatosis because of: 1) normal to low serum iron and transferrin saturation, without evidence of parenchymal iron overload; 2) the dominant transmission; 3) the lack of any relation with HLA; 4) the rapid development of iron-deficient anemia when unnecessary phlebotomies are performed. The molecular basis of the new syndrome is a mutation in the L-subunit ferritin gene on chromosome 19 (19q13.3-->19qter). The mutation involves a five nucleotide sequence [CAGUG] of the iron-responsive element (IRE), which is critical for the post-transcriptional regulation of ferritin synthesis by means of the binding with an Iron Regulatory Protein. As a consequence, ferritin synthesis is up-regulated, irrespective of cell iron status. PMID- 9417536 TI - [Cytomegalovirus pneumonia in patients with AIDS treated foscarnet. Our experience]. AB - A case of cytomegalovirus pneumonia occurring in a patient affected with AIDS is reported. The treatment of this disease is still difficult and remission of CHV pneumonia after therapy with foscarnet is emphasized. PMID- 9417537 TI - [Complete response to IFN in a case of typical HBV cirrhosis]. AB - A rare case of "wild-type" HBV cirrhosis (CHIL A/HBeAG+/HBV-DNA+) with complete response to IFN treatment after 3 successive series based on different types of IFN is reported. In this patient, HBeAg and HBV-DNA negativization after the second treatment with r-alpha-2b-IFN was observed and after the third treatment with lymphoblastoid-IFN HBcAb,-IgM negativization simultaneously with ALT persistent normalization. Over one year after the interruption of the last treatment, HBV clearance with HBsAg elimination and HBsAb, seroconversion was observed. The effectiveness of IFN was histologically confirmed with decrease of the piecemeal necrosis in the liver and presence of light fibrosis whereas the results of 3 previous histological evaluations showed: 1) CPH (1985); 2) CAH lightly active with initial signs of cirrhotic evolution (1988); 3) CAH with presence of nodular cirrhosis in the liver (1991). In particularly selected cases the possibility of a favourable response to the series of IFN treatment is stresses even in more advanced chronic "wild" HBV forms in which there is not evidence of mutants in the viral population. PMID- 9417538 TI - [A case of deep venous thrombosis and primary amyloidosis]. AB - On the basis of a complicated case of primary amyloidosis observed at our medical division, the clinical criteria and the biochemical and physiological supports correlated to this disease are discussed. In particular, it has been tried to make a distinction between primary and secondary amyloidosis. In this study, the clinical case of a 67-year-old man with a history of monoclonal gammopathy of uncertain significance, dizziness and syncopal episodes personally observed for a deep venous thrombosis of the inferior right limb is described. Of particular interest is the discovery of unusual resistance to anticoagulant treatment. PMID- 9417539 TI - [The new administration approach of the National Health System: implications for internal medicine. The point of view of the internist]. AB - The laws for the reform of the Italian National Health Service (502, 517) show marked bureaucratic and administrative patterns. The aim of these laws seems to be mainly a tight control of the health expenses instead of an improvement of health care. This choice is confirmed by the selection of indicators (DRG's) more fit for reimbursement rather than for quality assurance in health care. Moreover, these indicators are only a poor expression of the real medical status of the patients. The reform of the British NHS appears very similar to the Italian reform, mainly for the empowerment of managers to detriment of health professionals and of health care quality. The reforms designed with the collaboration of health workers may be more useful than the reforms based mainly upon bureaucratic principles. Only with this collaboration the double target of the reduction of health related costs and of the maintenance of an acceptable quality may be achieved. The professional associations and the academic world must be involved in the political and organisational choices related with health care, mainly in the fields of the setting of organisational standards and of the training of actual and future doctors. The model proposed is that of internal medicine as an example of the ideal set of skills useful for a mixed clinical and organisational task. The internists, used to the solution of complex clinical problems, may be the ideal candidates for the role of clinical managers. PMID- 9417540 TI - [Grepafloxacin (Vaxar)--innovative antibiotic for therapy of respiratory tract infections. Symposium: "Grepafloxacin--a new quinolone designed for respiratory tract infection. Lausanne/Switzerland, 25 May 1997]. PMID- 9417541 TI - [Discrimination against psychiatric patients in television films]. PMID- 9417542 TI - [Burden of relatives of chronic psychiatric patients]. AB - This study aimed at the systematic assessment of the burden imposed on the relatives of mentally ill persons. A postal survey was carried out on a sample drawn from the members of the Federal Association of Relatives of the Mentally ill (n = 557). RESULTS: Almost one third of those questioned experienced the care for their relatives as a heavy burden, a further third said that the burden was moderate. Among the negative consequences, health impairment was cited most often. Two-thirds of the participants reported constraints on their everyday life and the organisation of their leisure time. A (partly considerable) financial burden was cited with similar frequency. Further, the illness had a negative impact on the personal relationships of the caregivers. A quarter of the sample had lost touch with other family members. The same was found with regard to contact with friends. One-third reported discrimination by their environment. CONCLUSIONS: The results emphasize the necessity of creating opportunities to relieve some of the burden on relatives. PMID- 9417544 TI - [Accommodations for psychiatric patients. An empirical study of the Basel area canton]. AB - The psychiatric services of the Swiss region of Basel-Landschaft observed an increasing deterioration and shortage of accommodation offers for mentally ill persons. At the same time, it grew temporarily more and more difficult to let single rooms in flat-sharing communities. Therefore a psychiatric services research project aimed at evaluating the wishes and needs of specialists in the housing sphere, and of about 600 in- and outpatients, as well as for their caregivers. The results showed, similar to other studies, on the one hand patients' predominant wish to live independently, whereas on the other hand there was a great difference between the views of patients and their caregivers. The latter view this aspiration for independence rather skeptically. The striking disagreement between patients and caregivers may be fruitful for the therapeutic process; however, this applies only if both partners are aware of this fact. PMID- 9417543 TI - [Quality of life, needs and treatment evaluation of long-term hospitalized patients. Part II of the Berlin Deinstitutionalization Study]. AB - OBJECTIVE: Objective living situation, subjective quality of life, need for care, and assessment of treatment of long-term hospitalised patients were investigated. METHODS: 237 patients from 6 hospitals were examined using standardised instruments. RESULTS: Patients' satisfaction was generally relatively high, but varied greatly in subgroups and individuals. Patients were most satisfied with personal safety in the hospital and least satisfied with the expectation to live in the hospital for a long time. Therapists assessed clearly more needs for care in the patients than the patients did themselves. Statistical analysis revealed significant correlations between the subjective quality of life in different domains, the number of needs stated and plausible determining factors. CONCLUSIONS: Findings reflecting patients' living situation in the hospital are very heterogeneous. In respect of subjective evaluation criteria, there is little space for improvement in the future. PMID- 9417545 TI - [The suicide conference. An instrument for support of involved teams after inpatient suicides]. AB - This is a report on the authors' experience over two years in developing a new way to deal with patient suicides in a District Psychiatric Hospital. The main aim of the Suicide Counselling Session is to help the involved hospital ward teams to overcome the strain, doubt and psychological burden often caused by the suicide of a patient. The authors describe the institutional conditions and setting in which a Suicide Counselling Session takes place. The method is characterised by a special attitude and the ideas of "Reflecting Team" to bring the involved stress factors into the open. PMID- 9417546 TI - [Mobile crisis intervention and emergency psychiatry--3 years experiences]. AB - Three years ago in Linz the crisis intervention service started to offer mobile crisis intervention during the night and on weekends. The article presents concept, organisation of the service and statistical data. Although the crisis intervention service is responsible for a region with 330,000 inhabitants the utilization of mobile crisis intervention is low. PMID- 9417547 TI - [Development of forensic psychiatry (section 63 StGB) in North-Rhine-Westphalia. Comparison of the current situation with introduction of the forensic psychiatry regulation (MRVG-NW) 10 years ago]. AB - In a cross-sectional study we analysed the present situation of mentally disordered offenders treated in forensic hospitals of North Rhine-Westphalia under section 63 StGB-in comparison with the first block sampling in 1984. One of the important findings was that there has been an increase in violence in respect of the index delinquency as well as the criminal record. Those patients who were again committed to a forensic hospital showed more violent criminal offences too. The average duration of hospitalisation has been reduced from 6.1 to 4.8 years. PMID- 9417548 TI - [Compulsive disorder with homicidal impulses and paranoid symptoms in vascular encephalopathy]. AB - We report on the case of a 45-year old man with OCD who had the obsessive impulse to kill his 3-year old son. The patient showed signs of vascular encephalopathy after perinatal brain damage; besides that, he had developed a mild "explanatory" delusional system. Under treatment with SSRI and clozapin he improved remarkably. Presenting this case, we discuss the connection between organic disorders and OCD, and especially its relationship to perinatal brain damage. PMID- 9417549 TI - [The Gretchen question: acquittal after infanticide in denied pregnancy?]. AB - A mentally healthy woman killed her newborn directly after parturition. The physical and mental factors of pregnancy were continuously concealed, and even the menses persisted for nine months. Psychiatric expert opinion certified a psychologically exceptional state of mind of the woman, and the court acquitted her. PMID- 9417550 TI - [Comment on Nieder, J: Pre-admission and after-care--possible applications in psychiatry]. PMID- 9417551 TI - [Psychiatric-anthropologic field work. Sue Estroff: Making it crazy]. PMID- 9417552 TI - [Pharmacotherapy of chronic schizophrenic psychoses--neuroleptic plus serotonin reuptake inhibitor?]. PMID- 9417553 TI - [Psychiatric services in London]. PMID- 9417554 TI - ["High Noon"--course of an acute schizophrenic psychosis without psychiatry]. PMID- 9417555 TI - [Successful use of amantadine in treatment of malignant neuroleptic syndrome]. PMID- 9417556 TI - [The current status of implantable automatic defibrillators]. AB - The implantable cardioverter defibrillator has become an important therapy for patients with sustained or life threatening ventricular arrhythmias. Although the concept for the implantable cardioverter defibrillator originated in the late 1960s, the first device was implanted in humans in 1980. Since then, the technology has improved rapidly the design, function and reliability of the devices have been greatly modified. There are currently five companies dealing with defibrillators in Spain incorporating multiple options in defibrillation, pacing and sensing capabilities. New devices with atrioventricular pacing and atrial defibrillation possibilities will soon become available. The purpose of this article is to review the principal functions of implantable cardioverter defibrillators currently available. PMID- 9417557 TI - [4 proposals for ventricular remodelling in the surgical treatment of dilated myocardiopathy]. AB - Four surgical procedures are proposed to achieve an efficient remodelling of the ventricles with a low injury to heart muscle, for the treatment of the dilated cardiomyopathy. Those procedures are based the partial ventriculectomy technique of Batista an on the new conception of the macroscopical myocardium structure of the ventricles evidenced in the second half of the present century. PMID- 9417558 TI - [The thrombolytic treatment of acute myocardial infarct in an emergency department]. AB - INTRODUCTION AND OBJECTIVES: Although the importance of the early use of thrombolytic therapy in acute myocardial infarction has been demonstrated, it is usual to detect an unacceptable delay in its administration. We measured the in hospital delay and, when it was determined we designed a protocol to reduce it. METHOD: From January-92 to December-94 we performed a prospective analysis of the measured delay for patients with a diagnosis on admission of acute myocardial infarction or unstable angina within 24 hours of the onset of symptoms. To ensure a homogeneous population, we established a triage system: priority I, delay of the therapy not admissible and so immediate administration of thrombolytic agent (performed in the emergency department); priority II, need for a careful evaluation of the risk/benefit ratio for thrombolytic therapy and administration, when indicated, after admission to the coronary care unit, and priority III, thrombolytic therapy whether indicated or contraindicated. All data were evaluated periodically in order to detect possible failures and to correct them. RESULTS: A total of 1,462 patients with a diagnosis of acute myocardial infarction (n = 1,006) or unstable angina (n = 456) were included. The administration of lytic therapy in the emergency department reduced the In Hospital delay for thrombolysis by 54% from a median of 65 minutes (45 and 110) to 30 minutes (15 and 60) (p < 0.001) in priority I patients (40% of the patients diagnosed with AMI). For all cases with thrombolytic therapy this time was reduced from 87.5 minutes (50 and 155) to 50 minutes (25 and 110) minutes (p < 0.001). CONCLUSIONS: Awareness of our in-hospital delay, establishing a triage system in the emergency department and administering thrombolytic drugs in the this area has made it possible to provide this therapy to selected patients as early as possible. PMID- 9417559 TI - [Myocardial perfusion reserve studied by single-photon emission-computed tomography with thallium-201 and a drug stress test with adenosine triphosphate in patients with cardiovascular risk factors]. AB - INTRODUCTION AND OBJECTIVE: Recent studies have suggested that the evaluation of coronary reserve is a sensitive method in the early detection of vascular alterations before plaques exist, and certainly before clinical detection of atherosclerotic lesions. Single-photon emission-computed tomography (SPECT) with thallium-201 (201Tl) provides a noninvasive tool for evaluating myocardial perfusion reserve. The objective of this study was to measure the myocardial perfusion reserve in two groups of subjects, some with and some without cardiovascular risk factors and in a group of patients with coronary artery disease. METHODS: Seventy-four subjects, divided into three groups, were recruited to assess regional and global myocardial perfusion reserve. The control group consisted of 11 asymptomatic individuals without cardiovascular risk factors. The second group was composed of 49 patients with one or more risk factors. Finally, the third group included 14 patients with coronary artery disease. 201Tl-SPECT at rest and after pharmacological stress with a 7 minute adenosine triphosphate (ATP) infusion (140 micrograms/kg/min) was performed in all patients. ATP minus rest value subtraction was applied in order to obtain the stress data. Relative myocardial perfusion reserve indices were calculated as the ratio between stress and rest values. RESULTS: Global and regional myocardial perfusion reserves of the vascular territories were significantly lower in patients with cardiovascular risk factors than in control subjects (Global: 1.48 +/- 0.19 vs 1.81 +/- 0.08, LAD: 1.52 +/- 0.21 vs 1.85 +/- 0.09, CX: 1.45 +/- 0.2 vs 1.79 +/- 0.86, RCA: 1.47 +/- 0.2 vs 1.79 +/- 0.86) and higher than in patients with coronary artery disease (Global: 1.48 +/- 0.19 vs 1.31 +/- 0.14, LAD: 1.52 +/- 0.21 vs 1.35 +/- 0.15, CX: 1.45 +/- 0.2 vs 1.2 +/- 0.24). Univariate linear regression analysis in a group of 40 patients with high risk lipid profiles revealed a significant negative correlation between myocardial perfusion reserve and total cholesterol (r = -0.35; p = 0.01), LDL-cholesterol (r = -0.38; p = 0.036) and LDL/HDL ratio (r = -0.39; p = 0.029). CONCLUSION: Determination of myocardial perfusion reserve with 201Tl-SPECT allows the detection of abnormal vasodilatory response to intravenous ATP in patients with cardiovascular risk factors. These patients have higher reserves than patients with coronary disease, which might suggest an early phase of atherosclerosis. PMID- 9417561 TI - [The absence of interference between GSM mobile telephones and implantable defibrillators: an in-vivo study. Groupe Systemes Mobiles]. AB - INTRODUCTION AND OBJECTIVES: The electromagnetic field created by mobile telephones can cause pacemaker dysfunction. Although implantable cardioverter defibrillators are also susceptible to electromagnetic interference, few studies have addressed this issue and compatibility with the GSM mode has not been tested. This study was developed to detect possible "in vivo" interference between GSM mobile telephones and implantable cardioverter defibrillators. MATERIAL AND METHODS: The study group is composed of 30 patients with 8 different models of defibrillators. Twenty six had endocardial leads and 4 epicardial. Three GSM mobile phones were used: Siemens S3 COM and Motorola 6200 in all cases and Ericsson GA 318 in one. The tests were performed under continuous electrocardiographic monitoring. All therapies were deactivated and sensitivities were set to maximal parameters. The telephones were positioned in close contact to the defibrillator can and precordium, in two different angles. Three situations were evaluated: calling, established contact for 15 seconds and ringing. The protocol was repeated during pacing to assess the possibility of pacemaker mode inhibition. RESULTS: No cases of electromagnetic interference were observed. One patient presented non-sustained ventricular tachycardia episodes during the tests that were detected by the defibrillator. CONCLUSIONS: These results suggest that electromagnetic interference by GSM mobile phones are not a probable cause of implantable defibrillators dysfunction. PMID- 9417562 TI - [The heart in noncardiac diseases: how to decide in areas of uncertainty]. PMID- 9417560 TI - [Isotopic ventriculography in healthy young volunteers. Their response to different types of stress]. AB - BACKGROUND: Due to the increasing use of pharmacologic stress tests and the lack of comparative studies on ventricular function, this study was designed to establish the average limits in ventricular function with different kinds of stress, and to compare the response among them. METHODS: A randomized, open, controlled phase II clinical trial in 4 parallel groups was designed. Forty healthy male volunteers between 18 and 25 years were randomized into 4 groups of 10 individuals each: physical exercise (group 1), dobutamine (group 2), adenosine triphosphate (ATP) (group 3) and dipyridamole (group 4). Each volunteer underwent equilibrium radionuclide angiography, at rest and during stress. RESULTS: The global and regional ejection fraction increased significantly with the 4 kinds of stress. The maximal increase was reached with dobutamine and the minimal with dipyridamole. Physical exercise induced an increase in global ejection of 13 +/- 5%; dobutamine 16 +/- 6%; ATP 9 +/- 3% and dipyridamole 4 +/- 3%. CONCLUSIONS: The global and regional ejection fraction increases significantly more with dobutamine than with the other stress tests. Dipyridamole elicits the minimal increase. PMID- 9417563 TI - [Heart pathology of extracardiac origin (I). Cardiac involvement in AIDS]. AB - A great variety of cardiac disorders have been reported in HIV-infected patients: pericarditis, myocarditis, cardiomyopathies, endocarditis, cardiac involvement through malignancies, pulmonary hypertension, arrhythmias and thromboembolic disease. In general, these disorders are asymptomatic and often diagnosed in echocardiographic studies or autopsies. Pericardial involvement is the most common disorder. Pericardial effusions are asymptomatic and non-specific in a great proportion, but in some instances opportunistic infections or malignancies may lead to cardiac tamponade and are associated with an increased risk of mortality. The etiopathogenesis of myocarditis and cardiomyopathies is uncertain. There is controversy about the role of HIV as the primary etiologic agent. Opportunistic infections, cardiotoxic substances, nutritional deficiencies and autoimmune reactions have also been implicated as etiologic agents of myocardial damage. Short-term prognosis worsens as clinical manifestations of heart failure appear. Valvular involvement usually presents as marantic or infectious endocarditis, the latter most frequently in IVDU. This article reviews the main cardiovascular manifestations in AIDS. PMID- 9417564 TI - [The assessment of cardiac involvement in a case of a thoracic injury from a firearm]. AB - We report the case of a patient with a gunshot wound in the chest with a multiple small-caliber intrathoracic projectiles. The different noninvasive techniques employed to evaluate the anatomical location of these projectiles are discussed, together with their cardiac structural repercussions. The data provided by a simple chest X-ray, Computed Tomography (CT) and transthoracic echocardiography are commented on. A simple chest X-ray was unable to discern the location of the projectiles, in contrast to CT, which was able to identify both the number of projectiles and their location. The information provided was enhanced by transthoracic echocardiography, particularly in relation to those projectiles situated in anterior cardiac regions. PMID- 9417566 TI - [Conventional percutaneous transluminal coronary angioplasty as the form of treatment of complex intra-stent restenosis]. AB - We report the case of a patient with unstable refractory angina due to a coronary narrowing because of in-stent restenosis affecting the first diagonal, first septal and left anterior descending coronary artery. The lesions were successfully dilated with a conventional balloon catheter and a triple guide-wire system was placed through the inter-filaments space of the stent, to protect and recanalize the branch vessels involved. The patient evolved well and was discharged asymptomatic. PMID- 9417565 TI - [Recurrent syncope without angina: an uncommon presentation of coronary spasm]. AB - We report the case of a 64-year-old patient admitted for repetitive syncope as an isolated clinical manifestation probably due to coronary artery spasm. The patient had no history of previous cardiac disease, and was studied because of two nonspecific syncopes. Long-term electrocardiographic monitoring showed many episodes of transient ST segment elevation, associated with premature ventricular beats and runs of ventricular tachycardia. Coronary angiography during ergonovine infusion was performed to confirm the diagnosis. We discuss the incidence of coronary spasm provoking syncope and the need to establish a correct diagnosis in order to administer an effective therapy to the patient. PMID- 9417567 TI - [A false aneurysm with a double entry orifice]. AB - A case of a 64-year-old man is described, who was diagnosed as having a false aneurysm of the left ventricle, or pseudoaneurysm, a year following an inferior wall myocardial infarction. The echocardiogram demonstrated the presence of two ways of entry which communicated the inferior wall of the left ventricle to the false aneurysm. The patient was taken to surgery, where the diagnosis was confirmed. Aspects related to the diagnosis, treatment and prognosis of this case are discussed. PMID- 9417568 TI - [Does anesthesia always involve analgesia?]. AB - The primary aim of anesthesia is the prevention of pain perception (analgesia), intraoperative awareness (hypnosis), and postoperative recall (amnesia) (anesthetic trade). Analgesia would be a component of the anesthetic state. Pain is the conscious perception of a noxious stimulus and analgesia is the abolished perception of pain in an otherwise conscious patient. During general anesthesia pain can not be experienced, since the patient is unconscious. However, there is no certainty that during anesthesia a blockade of the nociceptive system is reliably provided in all circumstances. The impact of insufficient blockade of nociceptive pathways during anesthesia on mental status and implicit memory are largely unknown. Therefore, the anesthetic trade should consist of hypnosis, amnesia and antinociception. PMID- 9417569 TI - [What is your diagnosis? Alcoholic cardiomyopathy and chronic bronchitis]. PMID- 9417570 TI - [Contraceptive agents and risk of thrombosis]. AB - In the late sixties and seventies, publications of the Royal College of General Practitioners in England reported that in women using oral contraceptiva the incidence of venous thromboembolism is increased by two to four fold. Moreover, it was demonstrated, that these alterations in coagulation were induced by ethinylestradiol in a dose dependent manner. Following these findings, its dosage was lowered from more than 100 micrograms to 20-30 micrograms per day. More recently, the role of gestagens in inducing thrombosis has also been debated. Different authors observed an increased risk for venous thromboembolism in women using third generation pills containing gestoden or desogestrel compared with users of second generation levonorgestrel contraceptiva. These reports have generated a lot of concern and fear in the patients as well as doctors and have led to a drastic fall in the use of oral contraceptives. Due to the unavailability of safe contraceptive alternatives, the number of women experiencing unwanted pregnancy and its complications increased significantly. Indeed, direct proof for the role of gestagens in inducing thromboembolism is still lacking as the protocol designs of these studies do not allow us to infer whether the effects are due to the gestagens or to confounding variables. Hence, the discussions were beneficial for clinicians to remember the importance of checking the patient for individual and family risks for thrombosis before handling out a pill prescription. PMID- 9417571 TI - ["Zurich Vertigo Meeting"--phobic postural vertigo]. AB - Phobic postural vertigo has been described as a syndrome that is distinguishable from agoraphobia, acrophobia, and "space phobia". Closely related to locomotion, it is characterized by a combination of nonrotational vertigo with subjective postural and gait instability mainly in patients with an obsessive-compulsive personality. The monosymptomatic disturbance of balance manifests with superimposed attacks that occur with and without recognizable provoking factors in the same patient and are experienced with and without accompanying excess anxiety, misleading both patient and physician to a false diagnosis of organic disease. PMID- 9417572 TI - [Cortical vertigo]. AB - Dizziness of cortical origin is the subjective correlate of a disturbance of spatial orientation resulting from cerebrocortical dysfunction. Cortical dizziness in the form of vertigo is rare. If present, it most probably reflects a dysfunction of a vestibular representation in the insula. It may be accompanied by tinnitus, sensory disturbance and possibly also spontaneous nystagmus. The dysfunction of this region may result either from a focal seizure or from a lesion, for instance due to ischemia. Nondirectional, visual dizziness is most probably much more common than vertigo. This latter type of dizziness results from a functional disturbance of those parts of parietooccipital cortex, contributing to the discrimination of self-induced and externally-induced retinal image slip. It is not accompanied by additional symptoms and should immediately cease upon closure of the eyes or avoidance of ego motion. PMID- 9417573 TI - [Digoxin reduces morbidity, but not mortality in patients with heart failure]. PMID- 9417574 TI - [Amnesia. Transient global amnesia]. PMID- 9417575 TI - [Intense abdominal pain without signs for peritonitis. Acute mesenteric ischemia cause by embolism occlusion of the superior mesenteric artery]. PMID- 9417576 TI - [What is your diagnosis? Post-traumatic amnestic episode]. PMID- 9417577 TI - [New concepts in preventing human error in the operating room]. PMID- 9417578 TI - [Chronic diseases, coping with illness and complementary medicine]. AB - The increasing demands for complementary medical practises by patients with chronic diseases is seen in the context of a global model of contending with disease. The author assumes that only few variables of this model may influence "supply and demand" in the "marketplace of complementary medical methods" and that the importance of these methods may fade. Finally the encounter with patients asking for complementary medical methods is considered. PMID- 9417579 TI - [Complementary medicine--health policy and health care costs]. AB - Complementary medical methods are increasingly offered by physicians as well as other members of the health services and numerous therapists and they are sought and used by a majority of the population. From the point of view of health care authorities the question of acceptance or admission of methods and offering persons is raised. Unfortunately Switzerland lacks an uniform federal curriculum for practitioners of natural cures of other therapists in complementary medicine. Therefore, in order to control the steadily increasing wild growth of complementary medical offerings, several states (Kantone) have introduced examinations for the registration of such paramedical practitioners. Whether complementary (paramedical) medicine will really reduce costs of health care remains unproven so far. According to several surveys most alternative methods are used in addition to "school medicine" (academic medicine). Health insurance companies should subject reimbursement of costs to the fulfillment of standards regarding efficacy, appropriateness and economy equal to all other medications and therapies. PMID- 9417580 TI - [Paracelsus and sectarian movements in medicine]. AB - During his entire life Paracelsus used to criticize sectarianism in medicine. All trends, even the scientific ones, incur the danger to develop into sects, namely when they consider solely one cause for diseases and one treatment. Paracelsus often filled traditional prescriptions and used traditional methods, however, he worked as well in part with early homeopathy, magnetopathy and with magic and spiritual healing. Even wonders were not excluded. This all corresponds to an open, realistic medicine. Sectarian behaviour begins, however, when any of these trends is considered the only effective one. Most important in the medical philosophy of Paracelsus is the admission of the limitations of medicine and of all its trends. Through this scientific humility the physician knows that palliation in seemingly or really hopeless situations is one of his noblest tasks. PMID- 9417581 TI - [Critical evaluation of a "therapeutic study"]. PMID- 9417582 TI - [Severe soft tissue infection with bulla formation, sepsis, multiple organ failure]. AB - A 53-year old patient with diabetes mellitus presented with a painful swelling of the left thigh in an out-of-town hospital. Because of the slow progression associated with multiorgan dysfunction, a soft-tissue infection with sepsis was suspected. Strains of streptococcus of serogroup B were isolated from specimens, taken from certain areas of the affected skin. After transferring the patient to our hospital the diagnosis of a B-streptococcal associated necrotizing fasciitis was confirmed. Despite intensive medical treatment and several surgical interventions the patient deceased due to an acute severe liver failure as a consequence of a secondarily developed septic shock. In comparison to the well known cases caused by streptococcus group A, the B-streptococcal associated necrotizing fasciitis is a rather rare occurring disease. PMID- 9417583 TI - [Back and abdominal pain after minor injury in sprue]. PMID- 9417584 TI - ["Whiplash" injury of the cervical spine: value of modern diagnostic imaging]. AB - "Whiplash injuries" are frequent sequelae of motor vehicle accidents. While conventional imaging methods such as X-ray, including special and functional lateral projections, continue to be the first-choice evaluation methods, they frequently do not reveal injuries to the soft tissues. Cross-sectional methods such as CT and MRT may therefore be indicated for further workup in given clinical situations. In this paper the role of the entire spectrum of imaging methods is reviewed. Emphasis is placed on so-called functional CT, which allows detection of rotational instabilities of the upper cervical spine. These are difficult to evaluate clinically or by standard imaging studies, yet they may be a frequent cause of chronic whiplash syndrome. Furthermore, some results of morphologic studies of the alar ligaments by MRT are presented. Although experience to date is limited, it is likely that MRT of the occipitocervical junction region due to its potential to reveal exquisite anatomical detail, will gain in importance in the future. PMID- 9417585 TI - [Stereotaxic radiotherapy of brain metastases: experiences in Lausanne]. AB - We report the first series of patients treated by stereotactic radiation therapy for brain metastases in Switzerland. From August 1993 to December 1994, 19 patients were treated using a linear accelerator adapted for stereotactic treatment set-up. Most of the patients received combined treatment including external irradiation. The crude overall control rate for brain metastases was 79%. Median survival was 12.2 months. Overall survival at one year was 50 +/- 12%. These data compare reasonably well with the reported median survival of 3-6 months using external cranial irradiation alone. There was no "late toxicity". The data thus indicate that stereotactic radiation therapy combined with external whole brain irradiation appears to increase life expectancy and quality of life in a selected cohort of patients. PMID- 9417586 TI - [Neurology for practicing physicians]. PMID- 9417587 TI - [Vertebrobasilar insufficiency]. AB - Vertebrobasilar insufficiency is a very imprecise and much too frequently applied diagnostic term, usually used to classify uncharacteristic intermittent symptoms such as dizziness or visual blurring. Detailed history may make it possible to identify more specific details. Diagnostic attempts nowadays should aim at identifying underlying pathophysiologic mechanisms: intrinsic vessel disease (e.g. atherosclerosis), extrinsic vessel compression (e.g. cervical spondylarthrosis), systemic affections (e.g. anemia, hypotension), or combined etiologies. Accordingly, various therapeutic options exist. Ultrasound examination of the vertebrobasilar system is a reliable screening method in skilled hands, though its sensitivity is lower than in carotid artery disease. To identify the site and nature of the underlying pathology, digital angiography and, probably soon, magnetic resonance angiography (MRA) are available. PMID- 9417588 TI - [Advances in therapy and prevention of ischemic myocardial infarct]. AB - In the acute stage of stroke, fibrinolytics are beneficial for up to 3, and in some patients up to 6, hours. If fibrinolytics are contraindicated, aspirin should be given. Heparin is dangerous due to the threat of intra- and extracranial hemorrhage. For secondary prevention, anticoagulants are indicated. When hypercholesterolemia is present, statins should be given. PMID- 9417589 TI - [Tension headache and migraine--management in general practice]. AB - Tension type headache, migraine, and chronic daily headache as a possible endpoint of both, are the most frequent manifestations of primary headache. Their effects on the quality of life and at the workplace, as well as the socioeconomic consequences, are largely underestimated. The aim of treatment is the elimination of the condition or at the very least an acceptable improvement for the patient. If self-help fails, a professional approach is needed, priority being given to exclusion of symptomatic secondary headache and the necessary diagnostic classification in order to initiate treatment. These are primarily general measures, although often medication is indispensable. Empathic use of current knowledge, employing optimal means and methods, offers the best chance of success. PMID- 9417590 TI - [Neurosurgical treatment of Parkinson disease and other movement disorders]. AB - The surgical treatment of movement disorders has attracted widespread attention in the past few years. Better understanding of pathophysiological mechanisms, and also the refinement of neurosurgical, neuroradiological and neurophysiological methods and techniques have contributed to the redefinition of surgical treatment. With appropriate selection criteria, symptomatic and functional benefit can be achieved in the majority of patients. Functional neurosurgical procedures nowadays are performed most frequently in patients with Parkinson's disease or cervical dystonia. Functional neurosurgery today has the choice of a variety of different surgical options. The most appropriate operative technique is chosen with regard of the clinical presentation of the individual patient. The different operative procedures and the postoperative results are briefly described. PMID- 9417591 TI - [Renal colic and flakes in the urine]. PMID- 9417592 TI - [Ultrasound findings of celiac trunk stenosis in real-time B image]. AB - Ultrasonography, especially real-time-B-mode, is outstandingly suitable for detecting larger alterations of abdominal blood vessels or stenosis of the truncus coeliacus. This stenosis could be detected in 18 cases by real-time scanning. 10 of these diagnosis were confirmed by angiography. The clinical value of these findings is discussed. PMID- 9417593 TI - [Ultrasound measurement of gastric emptying time values]. AB - Sonography, as a non-invasive procedure, may be used also for studying the gastric emptying time. After 250 ml of a liquid test meal the gastric emptying time was determined in 20 subjects. In each subject the applied test meal could be traced in the prepyloric antrum. When examined at 8.00 a.m. the fluid had left the stomach after an average time of 8.3 min. During the day the emptying time increased to 10.8 min. A second examination on another day after application of 30 mg metoclopramide revealed significantly lower gastric emptying time. Ultrasonography may therefore be used as a sensitive non-invasive method for studying the gastric emptying rate in the fasting state and under pharmacological stimulation. PMID- 9417594 TI - [In vitro and in vivo imaging of puncture instruments in the real-time ultrasound image. 1. Puncture needles]. AB - The article reports on systematic sonographic examination of needles for aspiration biopsy in a waterbath and in an animal model. The in-vitro findings were found to be applicable to the in-vivo situations. Needles can be visualized almost true to construction in horizontal longitudinal view and produce an angle like reflex at the tip in oblique longitudinal view. Roughening of the needle surface yields an amplification of contrast in the sonographic image. The same effect may be obtained by roughening the intraluminar parts of the needle. This is achieved by the corkscrew turns of the Nordenstrom needle or by means of stabilized air bubbles in gel form, the latter being interpreted as a kind of contrast substance. Internal inhomogenerization while maintaining a smooth exterior surface leads to contrast augmentation of the needle in the sonographic image without surface-induced traumatization during puncturing. PMID- 9417595 TI - [Ultrasound and pancreatic length]. AB - In 50 patients subjected to ultrasonic examination the length of the visualized pancreas was determined. The mean length was 8 cm. The whole pancreas could not be imaged in any of the patients. Further efforts will be necessary to visualize the pancreas to its full extent. PMID- 9417596 TI - [Measuring blood flow velocity, blood volume flow and arterial diameter using integral Doppler ultrasound--comparison and synthesis of 2 solutions]. AB - Starting with a short comparison of the space resolving, pulsed Doppler method with the integral Doppler techniques for blood flow measurement, the formula is derived, after which the profile independent cross-sectional mean velocity v is to be measured using the integral method. When evaluating the Doppler signal power, moreover, volume flow Q can be measured. Two versions of the integral method for volume flow Q, one with (WAVUD) and the other without (WUVUD), and the need for a separate determination of the angle of incidence are explained and compared. Combination of the two results in a new method for measuring the mean velocity v and cross-sectional area F of blood flow from one single sound direction, without the need for determining the angle of sound incidence into the artery. The main problems of the integral method are addressed and in vitro test results are reported, showing that 20% measurement accuracy should be practical in vivo. PMID- 9417597 TI - [An axicon lens for focussing ultrasound fields]. AB - In this paper the properties of an ultrasonic axicon lens of 10 cm diameter are Investigated. This device generates a narrow ultrasonic beam in a range of 20 cm. The beam width varies from 1.7 to 0.5 mm in the frequency range from 1.7 to 5.5 megahertz. For practical applications a water path is necessary. PMID- 9417598 TI - [Ultrasound detection of free air in the abdominal cavity]. AB - Diagnosis via x-ray film had so far been the only way to prove the existence of free air in the abdominal cavity and hence the presence of gastrointestinal perforation. However, it is possible to establish the existence of free air in the abdomen sonographically provided a suitable examination method is employed. A characteristic feature is represented by pronounced pre-hepatic echoes with multiple echoes or sound shadow phenomenon. Experimentally it was possible to identify 1 ml air in the abdomen of an ascites patient both by x-ray examination and by sonography with equal certainty. The clinical usefulness of the ultrasound method was scrutinized. Of 10 patients with gastrointestinal perforations, 9 were recognized by sonography, including roentgen-negative cases; in one case only, sonography yielded a false negative result. Sonographic differential diagnosis is more versatile and differentiation can be practised by the highly experienced investigator only. The proof of the existence of free air in the abdomen represents a considerable extension of the scope of sonographic emergency-case diagnosis. PMID- 9417599 TI - [Amebic liver abscess: ultrasound and clinical follow-up of 20 patients]. AB - A retrospective analysis of 20 patients with amoebic liver abscesses was performed. Seven of 20 patients had more than one abscess. In all, 33 separate abscesses were observed. The diagnosis was based on clinical findings such as malaise, fever, abdominal discomfort, and pain in the right abdomen, haematological and serological anomalies, such as leukocytosis, elevated erythrocyte sedimentation rate, and slightly elevated liver enzymes. The serological investigation included complement fixation, indirect haemagglutination, and latex fixation test, this latter test produced strong reactions in all patients with acute symptoms. The typical ultrasonographic pattern resembled that of a minor echogenic lesion at the time of diagnosis and initiation of therapy. However, in a few acutely ill patients the lesions could not be identified by ultrasonography on first examination, but were clearly identifiable on second investigation on the following days. This finding has not been published previously. It may be observed in a very early stage of the disease only and is, therefore, dependent on rapid diagnosis of amoebiasis. During the follow-up investigations of three months to one year, the majority of abscesses could still be identified. PMID- 9417600 TI - [Training and quality assurance in ultrasound diagnosis]. AB - Medical ultrasound equipment is mainly used for diagnostic purposes. Examiner, patient, measuring equipment and ultrasound-based diagnosis form the ultrasound diagnostic system which is characterised by a multitude of internal and external relations. By extending the scope of the system, quality assurance of the ultrasound diagnostic procedures is improved with regard to the physical performance of the system. The performance standards should be established internationally. Even the best of equipment is only as good as the user's experience In this respect, the standards have to be met theoretically and practically on all levels of medical education. The most appropriate settings would be represented by national associations of ultrasound. The introduction of ultrasound diagnostic procedures on the level GP everyday work should lead to a decrease in cost and an increase in speed and efficiency by eliminating routine x ray examinations. PMID- 9417601 TI - [Prerequisites for offering ultrasound services in general practice]. AB - In the course of the past few years, sonography has become firmly established in clinical practice. The accuracy of the investigations does not only depend on the quality of the equipment used but also to a considerable degree on the standards of investigator training. To reduce the discrepancy between the number and quality of the investigations in the German Federal Republic, the "Kassenarztliche Vereinigung" has released guidelines requiring the users of sonographic equipment to undergo theoretical and practical training. These guidelines are reviewed and commented. PMID- 9417603 TI - [Thoughts on training in ultrasound diagnosis of brain-supplying arteries]. AB - Training programmes for the ultrasonic investigation of the arteries supplying the brain should be primarily oriented towards exercise and acquirement of skill in handling the probe. This is true for both Doppler sonography and real time B mode scanning. Postexamination reading of the stripcard recordings is not sufficient, and even with a more sophisticated data recording system, including frequency analysis and flow-imaging, the need for practical exercise remains, and the training period cannot be shortened. About 1000 examinations are required for sufficient training. Ultrasonic examinations of the carotid and vertebral arteries should be performed by neurologists or angiologists who are familiar with cerebral vascular disease. Only careful clinical evaluation assures the appropriate interpretation of Doppler findings. PMID- 9417602 TI - [Ophthalmologic ultrasound diagnosis--current training programs in Germany, Austria, Switzerland]. AB - In these three countries the application of diagnostic ultrasound in ophthalmology is restricted, with few exceptions, only to institutions having access to in-patients. This situation is based on the organization of medical care and does not depend on the technical evolution of the equipment. Consequently the annual rate of trainees in ophthalmic ultrasound is relatively low, but a comprehensive training programme is needed in this field. The requisite training subjects are described. They include a technical and a clinical programme for both pulse-echo (A, B, M, D mode) and Doppler techniques as applied for biometry as well as for tissue and vascular examination. Some data are given regarding training facilities, aids and courses in the three countries. The concept and organization of a 5-day course (Bonn/Wurzburg Course, Directors: W. Buschmann/H.G. Trier) is described in greater detail. For ophthalmic ultrasonography, testing and calibration of system parameters both for equipment and transducers is necessary for obtaining reliable and reproducible results. Examples are given for the organization of technical training in the practical course. The 1981 guidelines of the "Kassenarztliche Bundesvereinigung", Cologne, are discussed. These regulations define minimum requirements for both ultrasonic training and equipment in the FRG. Finally, a few controversial aspects of the teaching (quality assurance, role of biometry) are mentioned. PMID- 9417604 TI - [The contribution of ultrasound in urologic diagnosis]. AB - Urological diagnosis is considerably improved by the application of sonography. The best results are undoubtedly obtained if the investigations are carried out by an appropriately trained urologist familiar with clinical differential diagnosis. Various sonographic techniques which have proved helpful in this field are discussed, and a novel method for assessing the residual urinary volume is described. PMID- 9417605 TI - [Methods for discriminating flow direction with continuous wave Doppler equipment]. AB - The application of ultrasound Doppler units for the localization of incompetent venous valves, the examination of extracranial vessels, and the evaluation of the hemodynamic effect of certain cardiac dysfunctions requires directional models with the possibility of recording Doppler wave forms. Various techniques are used to determine flow direction all of them presenting specific advantages and disadvantages in practical use. After briefly explaining the basic Doppler technique, the three most commonly used methods, the two-channel filter technique, the McLeod system, and the outphaser system, are described. The characteristics of the various signal processing techniques are discussed, particularly in the case of complex hemodynamic conditions. PMID- 9417606 TI - [Experimental studies of signal fields of ultrasound broad band transducers]. AB - Quantitative analysis of ultrasonic signals, as increasingly practised, makes it necessary to increase the band width as well as the efficiency of ultrasonic transducers. An broad band ultrasonic transducer is presented which meets the requirements by double lambda/4 layer matching and was specially designed for medical ultrasonic spectroscopy. In an extended measurement programme transmitter characteristic, spatial dependence of the spectral content of the transmitter signals as well as beam geometry and its frequency dependence are investigated and compared with conventional transducers. PMID- 9417607 TI - [In vitro and in vivo images of puncture devices in real time ultrasound. 2. Plastic instruments]. AB - The article reports on a systematic sonographic examination of plastic puncture instruments in a water-bath and in an animal model. The plastic instruments used showed visualization of the lumen in a horizontal (longitudinal) cross-section. In an oblique longitudinal cross-section of an instrument filled homogeneously with water, only one echo generated at the tip of the instrument could be demonstrated. It was possible to achieve contrast enhancement of the plastic instrument by roughening its exterior or interior surfaces, by insufflation of air bubbles or by inhomogenization, using various plastic-cum-metal constructions. The effects produced in the water-bath were found to be similar to those observed in vivo. PMID- 9417608 TI - [Ultrasound diagnosis in Caroli syndrome--the method of choice]. AB - Ultrasonic exploration of the upper abdomen renders diagnosis of Caroli's syndrome easier, whereas the recognition of this condition has been very difficult so far. Secondary diseases complicated the course and resulted in unsuccessful surgery, since the underlying cause was not detected. It would be desirable to diagnose this congenital disease during adolescence by ultrasonic screening. Once the correct diagnosis has been made, it seems possible to considerably improve the otherwise fatal prognosis by means of an adequate drug therapy. PMID- 9417609 TI - [Ultrasound-controlled fine needle puncture of gastrointestinal tumors with continuous view]. AB - On account of its principle, ultrasound tomography cannot be recommended as a screening method for intestinal tumors; nevertheless, it may sometimes demonstrate larger tumors not producing typical signs. These neoplasms show a characteristic pattern ("cockade phenomenon"). In the region of the antrum they appear occasionally as a rigid thickening of the gastric wall. The nature of such an expansive lesion may be evaluated quickly and safely by a fine-needle puncture performed under sonographic control. No complications have been observed so far. PMID- 9417610 TI - [Comparative studies of various echomammography techniques]. AB - Until now it is not clear which echographic scanning technique is most effective and economic for breast examination. To answer this question we examined in a special study 189 patients with the following scanning units: 1. automated immersion scanning technique, 3 MHz (Octoson, Ausonics) 2. manual compound scanning unit (skin contact), 5 MHz (80 LDI, Picker) 3. linear array realtime scanner (skin contact), 5 MHz (SAL 20, Toshiba). Two examiners evaluated the image quality with regard to the visualisation of the histologically confirmed malignant lesions (n = 78). For testing the spatial resolution, we developed a tissue--mimicking phantom including cyst-like structures of different sizes. The clinical and technical advantages and disadvantages respectively of these three methods are discussed. PMID- 9417611 TI - [Clinical testing and improvement of an ultrasound method for detection of pathologic changes in the breast]. AB - Recent years have seen rapid advances in ultrasound techniques, increasing the chances of early diagnosis of changes in the female breast, irrespective of the presence of signs or symptoms. Sonography is excellent for the observation of breast tissue changes. However in common with other medical imaging techniques, it does not provide qualitative data--it is only a diagnostic guide. The article, therefore, includes a list of criteria which are useful in differential diagnosis of benign and malignant tissue changes. PMID- 9417612 TI - [Indications for ultrasound examination of the female breast--report by an established gynecologist]. AB - Experiences with sonography of the female breast are reported by a practising gynaecologist. Examples are described to show in what way sonography of the breast can be used to supplement x-ray film and thermography. Comments are made on the equipment required. The experiences cover a period of two years. Out of 2138 patients subjected to mammography, additional sonography was performed in 240 women. 28 carcinomas of the breast were detected. It is concluded that sonography is a meaningful additional method of examination to be performed complementary to mammography. PMID- 9417613 TI - [Real time ultrasound in routine breast diagnosis]. AB - Ultrasound examination of the breast is used in three indications: the premenstrual syndrome, the examination of cyst and the short-time control of high risk patients. The differential main point is the diagnosis of cysts, which may save the patient from "blind" puncture. In this field, ultrasound is more competent than mammography; furthermore, the oedematous changes occurring before the onset of menstruation can be easily seen with sonography. For the early detection of cancer of the breast, ultrasound is merely additional but its diagnostic value is bound to rise with progressing development. PMID- 9417614 TI - [Alarm signals in ultrasound evaluation of the gestational sac]. AB - The author stresses the importance of the echographic morphology of the gestational sac in evaluating pathological first-trimester pregnancies. The various ovular alarm signals are indicated and the abortion percentages are correlated to the various echographic patterns. The methods enables us to formulate a prognostic judgement which is both sufficiently valid and more closely consistent with the clinical reality. PMID- 9417615 TI - [Uterine foreign body and septic peritonitis after Cesarean section]. PMID- 9417616 TI - [M-mode and two-dimensional echocardiography of heart valve prosthesis dysfunction]. AB - The diagnostic efficacy of M-Mode and 2-d echocardiography for the detection of malfunctioning prosthetic valves was studied in twenty-nine patients with prosthetic valves and clinical deterioration. Echocaradiographic abnormalities were found in 8 of 12 patients with confirmed valve dysfunction; 30.8% of the cases had false negative echo tracings. In 17 patients with normal echocardiographic pictures of the valves another cause of clinical deterioration was found (congestive heart failure, pericardial effusion, aneurysm of aortic root). In this study M-Mode and 2-d echocardiography had an excellent diagnostic accuracy rate in distinguishing congestive heart failure from valve dysfunction. Comparing the data of M-Mode and 2-d registration, both methods are found to be complementary. A major advantage of the 2-d echocardiography is its enhanced spatial orientation, whereas M-Mode echocardiography enables better evaluation of valve movements and corresponding time intervals and recognition of quick movements and fibrillations. PMID- 9417617 TI - [Ultrasound diagnosis of pathologic changes of the uterine tube]. AB - The possibilities of sonographic diagnosis of pathologic changes in the uterine tube are examined by means of a computerised automatic real-time scanner with a statistical B-scan of high resolution. It is not possible to isolate sonographically the anatomically unchanged tube or the tube obliterated as a result of salpingitis. Nevertheless, it is possible to effect an indirect diagnosis if the tube is at least unilaterally patent or if it is bilaterally occluded, by means of an extended form of sonography within the scope of a sonographically controlled hydropertubation via a phenomenon which the author describes an "positive" or "negative" "phenomenon of Douglas". Hence, sonographically monitored hydropertubation, together with hysterosalpingography or laparoscopic chromopertubation--with reservations--is a valuable diagnostic tool to help in clarifying the cause of sterility. Pathological changes of the tubes in the sense of a sactosalpinx of different contents (retained secretions) can be differentiated by sonography. A characteristic sonographic echo pattern is also seen if the tubes are conglutinated with an extremely low amount of liquid collected in the tube. Various pathological changes of the tube can be recognised sonographically directly and indirectly by means of sonography extended in this manner. PMID- 9417619 TI - [Use of ultrasound in ophthalmology]. AB - In ophthalmology, ultrasound is applied in diagnostics as well as in surgery and therapy. This paper gives a short survey on both applications. Ultrasonic phacoemulsification is of considerable practical importance for modern cataract micro-surgery with intraocular lens implantation. Applications of that kind require consideration of ultrasonic bioeffects and equipment safety. Diagnostic use of ultrasound includes biometry (echometry), tissue examination and characterization, and vascular investigations in eye and orbit. The application of diagnostic ultrasound on in-patients, its individual indications, and the appropriate methods (A, B, automatic biometric devices for axial length measuring, M, Doppler) are described. Examples of commercially available instruments for the different applications are given. In comparison with other disciplines ophthalmic A-mode and B-mode echography is characterized by: refined depth resolution and lateral resolution; the important part of quantitative methods for clinical evaluation of echograms; and the advanced level of quality assurance for equipment performance. Refined tissue evaluation requires optimized and reproducible equipment parameters. To ensure these conditions the clinical echographer must be educated and willing to test performance and quality of his equipment. Finally, a perspective of actual research in diagnostic ultrasound of the eye is given. PMID- 9417618 TI - [Ultrasound mammography: development, current status and limits]. AB - Starting with a survey of the historical development of ultrasound-mammography the current status of this method within other diagnostic procedures of the breast is defined. Basing on over 1000 examinations since 1975 we report the sonographic possibilities in demonstrating different normal and pathologic breast structures. Pathologic structures are: Dudectasy, Cysts, Mastitis, Fibroadenoma, Carcinoma and Gynaecomasty. Own results are reported and the future of ultrasound mammography is discussed. PMID- 9417620 TI - [Current scanning and biometry equipment in ophthalmologic ultrasound diagnosis]. AB - The development of ophthalmological sonographic equipment started as early as 1958 with the ultrasonic "slit lamp" of Baum and Greenwood. This was an immersion type device with a closed membrane. Subsequent developments took place mainly after the introduction of the contact B-scanner by Bronson and Turner approximately 10 years ago. The instrument designed by them still serves as a source of inspiration for the creation of many "small-part" scanners. Among the special display modes featured in ophthalmological equipment, we have the deflection-modulated B-image (AB-mode) and the so-called "continuous-vector" mode for the A-trace selection from a real time B-mode image. Other examples are the specially designed A-mode equipment for measurement of the axial length of the human eye, and the A-mode equipment with a more or less standardized-gain characteristic curve of the overall amplification. PMID- 9417621 TI - [Developmental status and clinical significance of ultrasound diagnosis of intraocular diseases]. AB - Standardized echography is a special method of ultrasonic diagnosis developed for the purpose of ophthalmic tissue differentiation. It is a combination of A-scan 7200 MA (Kretztechnik) and a contact real-time B-scanner (Bronson-Turner). The special design and standardization of A-scan instrumentation and the wide range of application in the diagnosis of intraocular diseases are discussed and representative examples are given to emphasize the clinical significance of this method. PMID- 9417622 TI - [Axial echometry of the eye. Results and possible clinical applications]. AB - This survey describes ultrasound measurement techniques and echograms of living phakic and cataract eyes. Results of clinical echometry on emmetropic and ametropic eyes are reported. In unilateral or bilateral cataract and in unilateral aphakia, appropriate ultrasound measurements are of fundamental importance for restoring compatible vision; in unilateral aphakia, by means of combinations of contact lenses and spectacles; in cataract, by means of precalculated artificial intraocular lenses. PMID- 9417623 TI - [Differential echographic diagnosis in small tissue areas--exemplified by the orbits]. AB - Measurement-based ultrasonography proved mandatory in ophthalmic diagnostic work. It provides comparable examination conditions and therefore, comparable results, in contrast to simple, empirical ultrasonic examination. Measurement methods which can be easily applied under clinical conditions have been developed for determination of those technical characteristics of the apparatus and transducer probes which proved decisive for the diagnostic results. Some echographic criteria can be additionally or better evaluated using this basis. All echo amplitude measurements should be related to a well-defined test-reflector echo. In addition to sensitivity and resolution, frequency and frequency spectrum are especially important. Manufacturer's data have proved insufficient up to now, insufficient; even within one manufacturer's series of one equipment or transducer probe type considerable deviations from the declared data have been found. Such deviations may mimick pathologic alterations in the echograms. The size of a lesion area can be better evaluated when using well-defined technical conditions. The echographic presentation of tissue structures in the depth is especially dependent on frequency and on the frequency spectrum. Pathologic alterations of tissues may cause changes in the ultrasound attenuation which results in emphasized or reduced presentation of echoes from normal structures behind the lesion area. Tissue differentiation should be based on additional A scan echograms. Computerized echogram averaging provides a more reliable evaluation of echo amplitudes and ultrasound attenuation. Use of measurement based ultrasonography permits to compare measured echo-amplitudes and ultrasound attenuation with the results of other working groups, even if these are based on other equipment and transducers. PMID- 9417624 TI - [Value of echography for diagnosis and follow-up of perforating eye injuries]. AB - The introduction of vitreous surgery with its multiple capabilities of intraocular manipulations makes it possible to operate on cases which had been considered as hopeless up to now. With these techniques the importance of ultrasonic examination increased especially in cases with severe perforating injuries and their follow-up, to detect the onset of intravitreal fibrosis and early traction detachment, when no fundus view is possible. An other indication for ultrasonic examination is the exact localization of foreign bodies. Out of 484 perforating injuries which were examined and treated in our clinic during the last 10 years, ultrasonographic diagnosis was helpful in 18% and had further therapeutic consequences: In 24% a detachment operation or vitreous surgery was necessary, in 30% ultrasonography showed severe damage and surgery was considered to be ineffective. 42% were follow-up examinations of the posterior segment. PMID- 9417625 TI - [Ultrasound tomography and guided fine needle biopsy of intrathoracic space occupying lesions]. AB - Pleural and pleural-based intrathoracic masses lead to a real sound transmission, thus allowing a sonomorphological diagnostic approach. Thoracic sonography is particularly helpful to evaluate the nature of pleural-based radiographic opacities. Moreover, fine needle biopsies guided by sonography permit cyto histological diagnoses of intrathoracic masses. Ultrasound-guided fine needle biopsy was performed in 15 patients with pleural-based space-occupying pulmonary lesions. True positive results were obtained in 12 of the 15 patients with two true negative results and one false negative aspiration. The overall accuracy of aspiration cytology was 14 out of 15 (93.3%). In one patient a small pneumothorax occurred as a complication of the transthoracic needle approach. PMID- 9417626 TI - [Examination conditions for upper abdominal ultrasound in relation to diurnal rhythm]. AB - In 20 patients referred for sonographic examination, the conditions of examination in relation to daytime were evaluated. No impact of the conditions of examination in the abdominal region except for the region of the gallbladder was observed. Hence, sonographic examination of the abdomen should not be delayed if this investigation method is indicated. PMID- 9417627 TI - [Ascites of the fetus--intrauterine diagnosis and puncture treatment using ultrasound]. PMID- 9417628 TI - [Prenatal diagnosis of upper intestinal tract obstruction]. AB - Case report of prenatal sonographic diagnosis of obstruction of the upper gastrointestinal tract (atresia of jejunum). The typical echographic findings and the significance of the hydramnion which is regularly concomitant with this rather rare condition, are discussed. Prenatal diagnosis of a small-bowel tract atresia allows optimal neonatal management improving the prognosis. PMID- 9417629 TI - The NIH did it! PMID- 9417630 TI - The battle over BRCA1 goes to court; BRCA2 may be next. PMID- 9417631 TI - EMBL's outward expansion strains its core facility. PMID- 9417632 TI - Asian network applies a personal touch. PMID- 9417633 TI - Yale virus collection needs a home. PMID- 9417634 TI - Possible new roles for HOX genes. PMID- 9417635 TI - Pinning down cell division. PMID- 9417636 TI - Heavy ions pack powerful punch. PMID- 9417637 TI - Pieces of the true grail: a G protein finds its target. PMID- 9417638 TI - Getting around the nucleosomes. PMID- 9417639 TI - Temporal coding in neural populations? PMID- 9417640 TI - Balancing fish consumption benefits with mercury exposure. PMID- 9417641 TI - Crystal structure of the catalytic domains of adenylyl cyclase in a complex with Gsalpha.GTPgammaS. AB - The crystal structure of a soluble, catalytically active form of adenylyl cyclase in a complex with its stimulatory heterotrimeric G protein alpha subunit (Gsalpha) and forskolin was determined to a resolution of 2.3 angstroms. When P site inhibitors were soaked into native crystals of the complex, the active site of adenylyl cyclase was located and structural elements important for substrate recognition and catalysis were identified. On the basis of these and other structures, a molecular mechanism is proposed for the activation of adenylyl cyclase by Gsalpha. PMID- 9417642 TI - Kaposi's sarcoma-associated herpesvirus infection and multiple myeloma. PMID- 9417643 TI - Kaposi's sarcoma-associated herpesvirus infection and multiple myeloma. PMID- 9417644 TI - Kaposi's sarcoma-associated herpesvirus infection and multiple myeloma. PMID- 9417645 TI - Kaposi's sarcoma-associated herpesvirus infection and multiple myeloma. PMID- 9417646 TI - Kaposi's sarcoma-associated herpesvirus infection and multiple myeloma. PMID- 9417647 TI - [Therapeutic training aspects in rehabilitation after rupture of the anterior cruciate ligament in high performance athletes]. PMID- 9417648 TI - [General muscle rehabilitation practice]. AB - This article presents a five-step approach to the rehabilitation of sports injuries. (1) pain management; (2) specific exercise instruction for treatment of localized functional disturbances/motion restrictions; (3) training therapy to increase localized and general physical performance; (4) patient education and instruction of an individualized home/gym program and for precautions and injury prevention; and (5) enhancement of psychological performance. Described are the negative sequelae of a sports-related injury or immobilization after surgery on joints, bones and circulatory system, with particular emphasis of the effects on the involved musculature. The negative impact of an injury on generalized physical performance is presented (deconditioning syndrome). The course of rehabilitation requires a specific functional diagnosis, so as to implement an individualized treatment plan. Regular and objective follow-up/monitoring is suggested within the context of projected goals. The specific steps are described. Pain and discomfort can impede the restoration process and must be addressed effectively. Even though many questions remain to be answered in the field of muscular rehabilitation, the present day knowledge coupled with successful empirical interventions allow the introduction of effective and efficient rehabilitation concepts. PMID- 9417649 TI - [Development of joint mobility by rhythmic neuromuscular stimulation]. AB - According to Nasarov an outstanding improvement of flexibility could be reached by using the rhythmical neuromuscular stimulation (RNS). For proving this statement a study with 112 healthy fitness sportsmen and women was performed for three weeks at chest muscles. Overall, there was no clear advantage of RNS versus static stretch shown. However, it exists a dependence of improving flexibility by static stretch at simultaneous strength training. Persons who perform strength training obtain a higher improvement in flexibility than others. Using RNS the results in reaching high range of motions are independent of strength training and comparable to static stretch at simultaneous strength training. The results were discussed by a higher rate of biosynthesis induced by strength training respectively by the eccentric load of RNS. PMID- 9417650 TI - [Wheelchair basketball from the orthopedic viewpoint]. AB - 155 (aged 16 to 52 years) wheelchair basketball players were surveyed to determine athletic injuries and overload-syndromes. During their active participation in wheelchair basketball 60.6% of the players have suffered 272 injuries and overload-syndromes. Those were mainly localised at the upper extremity (74.6%). Acute injuries predominate with 57.7%. Strains of the finger joint and skin injuries were found to be the most common injuries. Myogelosis and tendinosis were the most common reported overload-syndromes. A significantly higher number of complaints were associated with wheelchair basketball participation at league-level and depending on the player's position. At the end of the article the means of possible additional disability and consequences to specify therapy and prevention for disabled will be discussed. PMID- 9417652 TI - [Changing physician's role]. PMID- 9417651 TI - [Electromyography-controlled muscle rehabilitation--knee injuries]. AB - Surface is a noninvasive, mostly nonreactive, objective measuring procedure for evaluation of the degree of muscle activation. It allows for accurate assessments of the neuromuscular system and description of adaptations resulting from injury. Decreases in torque, EMG amplitude and EMG frequency occur as a result of knee joint injuries. Efficiency checks and evaluations of training programs and neuromuscular treatment can be done. Decreases in torque, EMG amplitude and EMG frequency found after knee joint injury (cruciate ligament, meniscus) are due to intraarticular and periarticular afferent processes. 12 weeks postoperatively, the neuromuscular deficits from cruciate ligament injury have increased. While complete recovery can be achieved following meniscus fixation (more than 1 year postoperatively), cruciate ligament injuries lead to chronically reduced EMG median frequencies. PMID- 9417653 TI - [Living knowledge originates from practice]. PMID- 9417654 TI - [Injuries, women and violence in Bergen]. PMID- 9417655 TI - [Neurosurgery in the North]. PMID- 9417656 TI - [Ten years of neurosurgery at the University Hospital in Tromso. Need for further development]. AB - The Department of Neurosurgery at the University Hospital of Tromso was established in January 1986 to provide neurosurgical treatment for the population of northern Norway. During the first ten years, 3,225 patients were operated on, including 1,157 craniotomies and 1,335 spinal procedures. The annual number of operations increased from 201 to 442, and the number of hospitalized patients from 265 to 908. The number of patients treated per employee increased by 250%, while the mean duration of hospital stay decreased by 43% to 4.5 days. The growth in activity is expected to continue. This is because of improved diagnostics of nervous system disease, a growing number of elderly patients and new treatment options. The falling number of spinal surgical procedures in local hospitals is also a contributing factor. Neurosurgery is highly cost-effective. It is impossible to make further cost savings without a decline in quality. The department must be expanded to cater for more operations by increasing both the number of beds and employees. PMID- 9417658 TI - [Are life experiences of children significant for the development of somatic disease? A literature review]. AB - This review presents the relationship between serious life events, chronic family difficulties and illness, and focuses on how healthy children cope. Hospitalised children had experienced about twice as many serious life events as children in healthy environments. Known diseases related to stress are eczema, upper respiratory tract infections, asthma, ulcerative colitis, heart disease in adults, juvenile rheumatoid arthritis, fibromyalgia and juvenile diabetes. Research on healthy children at risk (resiliences) has revealed a number of social and interpersonal protective factors. A modified biopsychosocial model, for the purpose of understanding the health status and care of children at high risk, is presented. More research is needed to understand these multietiological diseases in order to develop strategies for the promotion of good health. PMID- 9417657 TI - [Women and violence. A one-year prospective study from Bergen emergency department]. AB - Medical studies on violence towards women have usually focused on domestic violence. Less attention has been devoted to other kinds of violence towards women regardless of cause. During a 12-month period in 1994-95 all victims of assault were recorded at Bergen Accident and Emergency Department. The attending physician completed a questionnaire on violence exploring where, when and how the assault happened, the use of weapons, whether the patient and perpetrator or both were under the influence of alcohol, and whether the patient intended to press legal charges. Patient characteristics and medical information were also recorded. 24% of the assaulted victims were females (241 of a total of 994). 131 of the females were victims of domestic violence, while 102 were injured in public places. Women injured in public places tended to be younger and more likely to be under the influence of alcohol than those injured by domestic violence. There were no significant differences in patterns of ICPC-diagnoses between the two groups, though the number of admissions to hospitals and referrals to specialists was higher in the group subjected to domestic violence. PMID- 9417659 TI - [Obsessive-compulsive disorders in children. A review based on recent research and clinical experiences]. AB - This article describes obsessive-compulsive disorder in childhood through a review of some recent research topics and the author's own clinical experiences with the disorder. Among the topics discussed are diagnosis, differential diagnosis, phenomenological characteristics and treatment recommendation. The article concludes that although recent research on obsessive-compulsive disorders in children has lead to important advances in our understanding and treatment of the disorder, much remains to be addressed. Research on childhood obsessive compulsive disorder in Norway seems to be non-existent, and the author calls attention to the need for empirical studies at Norwegian research institutions. These should include research on treatment preferences in Norwegian child psychiatry clinics. PMID- 9417660 TI - [Mental health among refugees. Connection between symptoms, traumatization and exile]. AB - Traumatization and psychiatric symptoms among 346 refugees admitted to the outpatient unit at the Psychosocial Centre for Refugees from 1992 to 1996 were registered and analysed by means of systematic, clinical interviews. More than 50% reported exposure to physical torture, and more than 50% had also been involved in serious war actions. Most patients had experienced many forms of persecution. Approximately 10% of the patients had employment in their host country and 20-30% attended school or participated in language courses. Almost 40% were neither employed nor engaged in any kind of studies. 50% were diagnosed as having post-traumatic stress disorder. The relationship between demographic background, traumatization and exile situation, and symptoms and social dysfunction is illustrated by case histories. PMID- 9417661 TI - [Diabetic care in Norwegian general practice. A report of current status from Salten and some regions in Rogaland]. AB - Few published data are available on the quality of diabetic care in Norway. This applies both to general practice and to hospital clinics. We reviewed the notes of 1,876 diabetic patients who were registered with general practitioners in Salten and Rogaland to assess the quality of care with reference to the Norwegian College of General Practitioners' guidelines for diabetic care. 89% of patients were classified as having type-2 diabetes. Hospital clinics were responsible for the care of 93 patients. Analysis of the results showed that during the last 12 months Hb A1c and blood pressure had been measured in 84 and in 86% of those patients under the care of their general practitioner. Some inspection of the foot had been carried out in 45% of the patients, and 37% of the patients had been referred to an ophthalmologist. Guideline targets for glycaemic control had been achieved in 46% of patients younger than 70 years of age (Hb A1c < 7.5%), and in 82% of patients older than 69 years of age (Hb A1c < 9%). Diabetic patients on insulin therapy had the worst glycaemic control. The study shows that the quality of diabetic care is not optimal, examination of the foot, referral for eye examination and glycaemic control of diabetic patients on insulin therapy are examples of areas where improvement is needed. PMID- 9417662 TI - [Evaluation of the General Practitioner Scholarship--small seed, rich harvest]. AB - In 1976 the Norwegian Medical Association instituted the General Practitioner Scholarship. A total of 36 scholarship months is granted each year, available for small scientific projects lasting a maximum of six months. This article presents the results of an evaluation based on data from the recipients of the scholarships (n = 107) and from rejected applicants (n = 76) during the period 1986-94, and from the four University Departments of General Practice in Norway. 85 (80%) recipients, involved in 95 projects, responded to a mailed questionnaire. 60 (72%) had published one scientific article or more from their project. 64 (70%) of the projects had evolved from questions raised in the recipients' own daily practice, the main topics being clinical issues and community medicine. More than two thirds of the recipients were still doing scientific work. All informants recognised the importance of the General Practice Scholarship for research and development in Norwegian primary health care. PMID- 9417663 TI - [Serum proteins in patients with recently discovered cancer in the oral cavity/throat. What experiences were gained by measurements of concentration and electrophoresis?]. AB - 84 patients with cancer of the upper digestive tract and 89 matched controls were examined. Lower than normal albumin concentrations were observed with an odds ratio for disease of 14 (95% confidence interval 4.5-46). Among the patients, low concentrations of serum total protein and albumin were also associated with a higher risk of dying. An essential finding was an electrophoretic serum pattern of "active process" without the production of gammaglobulins occurring in about half of the patients. This observation, when lasting for several months, seems to indicate a possible malignancy. However, one fifth of the patients had normal electrophoresis. On the other hand, 15% of the controls also had an "active process" pattern-with or without the production of immunoglobulins. With regard to the participants' patient-versus-control status, the electrophoretic serum protein pattern was of greater predictive importance than information concerning high or low erythrocyte sedimentation rate. PMID- 9417664 TI - [Memory clinic--ambulatory examination in suspected dementia]. AB - Dementia and confusion are frequently overlooked by general practitioners. A memory clinic was established at the Department of Geriatric Medicine, Ulleval Hospital in September 1990 with the intention of creating a standardized programme for diagnosing dementia and cognitive impairment in the elderly in an out-patient setting. The activities and services offered by the clinic are described: diagnoses, information to patients, their family and staff in the primary health care system about the symptoms and treatment of dementia. In addition, advice is given on follow-up care and how to apply for care through the national health service. There has been a great demand for the services of the Memory Clinic, and we believe that the methods used are adequate in an out patient setting. The concept could easily be transferred to other specialist clinics involved with the elderly, and parts of the programme could also be used by general practitioners. PMID- 9417665 TI - [Geriatric psychiatry--a specialty gaining recognition]. AB - Services for elderly, mentally ill people have developed in response to changing needs in society. In 1990 most of the 650 beds allocated to elderly patients in psychiatric hospitals were occupied by long-term care patients. Outpatient programmes hardly existed. In 1995 about 400 beds were allocated to geriatric psychiatry. They were served by 40 physicians and 20 psychologists. Out-patients' clinics were established. Most of the in-patients were short-term admissions. Nowadays, departments of geriatric psychiatry define themselves as diagnostic and short-term units. About a third of the in-patients suffer from dementia, a third from depression, and a third from various other psychiatric disorders. The authors recommend that a special unit for geriatric psychiatry should be established in every county in Norway. Funds should be allocated for professorships at all universities. PMID- 9417666 TI - [Geriatric psychiatry--a specialty within psychiatry]. AB - The aim of this article is to describe the prevalence of mental disorders in the elderly and how the psychiatric services for these patients ought to be organized in Norway. Geriatric psychiatry is a special branch of psychiatry. Its areas of concern are the assessment and treatment of mental disorders which frequently occur in the elderly. The most prevalent psychiatric disorders are depression and dementia. Functional psychosis and anxiety disorders are less prevalent, but nevertheless disorders causing great concern. Psychiatric morbidity frequently coexists with physical illness. An elderly patient suffering from a mental disorder often has a combination of psychological, social and physical needs. The resources allocated to psychiatric services for old people are scarce. Efforts should be made to establish a special unit for geriatric psychiatry in every county in Norway. Each unit should serve approximately 150-200,000 inhabitants, and should consist of both an in-patients' and an out-patients' clinic. It is recommended that there should be 1 to 1.5 beds per 1000 elderly aged 65 years and over. PMID- 9417667 TI - [Aging, dementia and automobile driving]. AB - According to Norwegian law, drivers 70 years and older must carry a health certificate. This is issued by a general practitioner. If the patient is not supposed to drive because of a medical condition, the doctor should report this to the County Health Officer. This can be problematic, not only because assessing whether a patient fulfills the criteria for driving is difficult, but also because the doctor has obligations to both the public and his patient. These problems are discussed, based on assessment of available literature and on personal experience. Dementia is common in old age and affects approximately 15% of persons aged 75 and older. Patients with moderate and severe dementia should certainly not drive. However, some patients with mild dementia can nevertheless be safe drivers. The problem, however, is to identify the safe drivers among patients with mild dementia. The current regulations on dementia and driving are presented briefly. PMID- 9417668 TI - [Who is in charge of health care development?]. PMID- 9417669 TI - [Alternative medicine]. PMID- 9417670 TI - [Does more frequent hair washing increase bone mass?]. PMID- 9417671 TI - [May suicide be prevented?]. PMID- 9417672 TI - [Psychocutaneous disorders in practice]. PMID- 9417673 TI - [Selective serotonin uptake inhibitors and adverse effects]. PMID- 9417674 TI - [Aging as scientific challenge]. PMID- 9417675 TI - [Drowning accidents--still a challenge for Norwegian physicians]. PMID- 9417676 TI - [Breast cancer]. PMID- 9417677 TI - [Vaccination against influenza--yes or no?]. PMID- 9417678 TI - [Breast cancer--incidence, mortality and stage shifting in Norway]. AB - The age-standardized incidence rate of breast cancer has increased by 50% over the period 1965-94. There has been a much lesser increase in the corresponding age-standardized mortality rate because of better treatment and stage shifting. Stage shifting means that the proportion of individuals with a given clinical stage changes over time. The proportion of individuals diagnosed as clinical stage 1 was seen to increase from about 50% to 60% in the time period mentioned above. The proportion classified as stage 2 at the time of diagnosis is constant at 30%. The relative numbers of individuals diagnosed as stage 3 or 4 were reduced from 10% to 3% and from 10% to 5%, respectively. After correction for confounding effect of age and the clinical stage, the age-standardized 3-year relative survival rate increased from 90% to 95% and from 67% to 85% for stages 1 and 2, respectively. The impact of advancing breast cancer diagnosis independent of the screening programme, is discussed. Finally, the evaluation of screening programmes using shift migration models and simulations is discussed. PMID- 9417679 TI - [Alcohol, breast cancer and causal inference in epidemiology]. AB - In order to demonstrate how possible causal relationships are critically evaluated in epidemiologic research, literature on the association between alcohol and breast cancer is reviewed and discussed. A cause can be defined as a factor which, in combination with other factors, known and unknown, is sufficient to produce an effect. Since the hypothesis-generating study was published in 1977, a total of 34 positive and 16 negative studies have been published. Methodological problems, such as chance, bias and confounding, cannot be considered as plausible explanations for the above majority of positive findings. The question of causality was then evaluated using the guidelines developed by Bradford Hill in 1965. Among these, the strength of the association, consistency, temporality, biological gradient and biological plausibility, are the most important. In spite of the relatively weak association and somewhat inconsistent results, it is concluded that alcohol consumption should be considered as a cause of breast cancer. It is estimated that in Norway, between 24 and 180 cases of breast cancer may be attributed to alcohol consumption. Future research should focus on the question of effect-modification and on the possible implications of different patterns of alcohol consumption. PMID- 9417680 TI - [Primary invasive breast cancer in Oslo 1980-89. A population based study of 1942 unselected patients treated by radical surgery]. AB - A retrospective review is presented of 1,942 consecutive patients with histopathologically confirmed invasive breast cancer, who where treated radically in Oslo from 1980 to 1989, either at General Municipal Hospitals (MH) or at a national Comprehensive Cancer Center (CC). The treatment and outcome were related to accepted medical parameters and the responsible hospital category. The median number of axillary lymph nodes described at the MH was 8, compared to 14 at the CC. During the period 1984 to 1986, 62% of the patients had an estrogen receptor analysis (MH 60%, CC 85%). Of the patients with N > or = 4, 36% received adjuvant hormone treatment (MH 32%, CC 67%), whereas the comparable percentage of patients with pT3/pT4 was 53%. The five- and ten-year cancer-related survival rates were 83% and 71% respectively, with no difference between the MH and the CC. As nation wide criteria for the management of breast cancer have been accepted, compliance and monitoring by surgeons, pathologists and oncologists is imperative as part of quality assurance. PMID- 9417681 TI - [Breast cancer treated at the oncologic department, University Hospital in Tromso 1986-94]. AB - In the period 1986 to 94, 173 women who had had a lumpectomy or a mastectomy were treated with radiotherapy at the University Hospital of Tromso. The median diagnostic delay was 2.4 months (range 0-98.6 months). Three out of four patients were operated on within two weeks of the diagnosis being made. About two thirds experienced a delay of more than six weeks from the operation to the start of radiotherapy treatment. The five-year overall survival rate in the mastectomy and postoperative radiotherapy group was 67%. Patients with estrogen receptor positive tumours had a better prognosis. Only 5% and 7% of all patients in our region in stages I and II had breast conserving surgery (BCS) during the study period (66 patients). The five-year overall survival rate in the BCS group was 77%. BCS raised the cost per patient by about 3,000 GBP compared to modified radical mastectomy (MRM). The cost per QALY using BCS as against MRM was about 12,000 GBP. We conclude that MRM should not be used instead of BCS merely for economical reasons. PMID- 9417682 TI - [Interstitial laser coagulation in the treatment of benign prostatic hyperplasia. Preliminary results]. AB - 51 patients aged 68 (range 52-81) years with lower urinary tract symptoms compatible with obstruction from benign prostatic hyperplasia were treated with interstitial laser coagulation (ILC). Postoperative urinary retention lasting less than one week was seen in the majority of cases. All patients were followed up for three months and ten cases had further follow-up after one year. Three months after treatment the international prostate symptom score decreased from 23.3 +/- 0.7 to 8.9 +/- 0.8 and was 10.2 +/- 2.1 after one year. Peak urinary flow increased concomitantly from 8.3 +/- 0.4 to 12.2 +/- 0.7 at three months and was 11.5 +/- 1.4 ml/sec after one year. Three patients received other, additional treatment because the ILC-treatment failed. In conclusion, interstitial laser coagulation had marked effects on symptoms, whereas the effects on objective parameters were less pronounced in this selected group of patients. However, more extensive follow-up, is essential for further evaluation of this new treatment procedure. PMID- 9417683 TI - [Colorectal cancer. Location dependent symptoms?]. AB - It is claimed that the symptoms of cancer in different parts of the colorectum are varied. The aim of the study was to decide how well the "classical" symptoms are divided between left and right colonic and rectal adenocarcinomas. This was a retrospective study of 102 patients with colorectal cancer at Ulleval hospital in 1992. Red blood in the faeces (p = 0.001), and changes in stool pattern (p = 0.001), were left colonic and rectal cancer specific. Moreover tenesmus, mucous stools, and pain at defecation were specific of rectal cancer. All these findings agree with the textbooks. Melaena, diarrhoea (p = 0.23), weight-loss (p = 0.09), a feeling of general physical weakness (p = 0.13), and ileus/subileus were not found as localisation specific symptoms, contrary to the claims of several textbooks. Pain was always correlated to the affected side in left and right colonic cancer, while in rectal cancer the localisation of pain was non-specific, and it appeared rather often; (37%) in Dukes' A and Dukes' B. Only a few patients had anorexia, but the number was still significant for those with right colonic cancer (p = 0.04). With few exceptions, our results generally support the textbooks. The referring symptoms may help to decide the necessity for further examinations, when total colonoscopy is not obtained. PMID- 9417685 TI - [Rehabilitation of discharged patients is beneficial. Medical and economical analysis]. AB - Even after the Swedish "Adel"-reform in 1992, large Swedish emergency hospitals still have many "bed-blockers" waiting to be discharged for further care by the social services. In Malmo a special ward, run by the social services, was created in 1994 for the purpose of treating, rehabilitating and planning the discharge of "bed-blockers" from the university hospital. The "bed-blockers" treated at this ward during an 18 month period (patient group, n = 223) were compared with a control group of "bed-blockers" (n = 285) who were not admitted to this ward. Although the patients in the patient group seemed to be more sick and handicapped than those in the control group, emergency in-hospital care during a follow-up period of one year was four times greater in the control group than in the patient group. Economic analysis showed that rehabilitation of the patient group was cheap and effective. The study has shown that municipal care of "bed blockers" needs to be improved and better organised. PMID- 9417684 TI - [Prognosis one year after hip fracture]. AB - This study evaluates the demography and health in a hip fracture population, and predictors of outcome one year after the fractures occurred. Physical, mental and social functioning in 109 patients who were referred from home with hip fractures were assessed retrospectively; during the hospital stay, at discharge, and after 4 and 12 months. Mobility, Katz' ADL-index and a short-version of MMSE were used in assessing their physical and mental conditions. No essential changes were found in either demography or health. The most significant predictors of outcome were age, prefracture mobility and post-fracture mental status. The proportion of patients suffering from acute confusion was considerable. The result was a higher risk of mortality, institutionalisation and poor physical outcome. It is important to pay more attention to the prevention and treatment of cases involving acute confusion. PMID- 9417686 TI - [Chromogranin A in blood--a useful tumor marker]. AB - Chromogranin A (CgA) is a protein localized to the secretory granules of neuroendocrine cells. Determination of CgA in blood is a useful marker for neoplasia and hyperplasia of neuroendocrine cells. Chromogranin A in serum should be the first test used for detecting carcinoids. It is also useful for diagnosing other neuroendocrine tumours, such as oat-cell carcinoma of the lung, pheochromocytoma and neuroblastoma. PMID- 9417687 TI - [Measurement of health-related quality of life in malignant diseases. Does it have any consequences?]. AB - The traditional end points of clinical trials in malignant disease are response, disease-free survival, and overall survival. Because a substantial part of cancer treatment is palliative, health-related quality of life should be a major objective in clinical trials. Cancer specific questionnaires, which measure the various quality of life domains and which are tested for reliability, validity and sensitivity to changes in clinical status, are now available. Measurement of health-related quality of life may lead to important conclusions. For instance, results from a Nordic myeloma trial suggest that obtaining an objective response is not a prerequisite for achieving an improved quality of life. They also indicate that the 5 to 6 month plateau phase prolongation obtained by giving interferon alpha-2b in addition to melphalan/prednisone is not associated with any quality of life benefit. Data from the same trial are used to illustrate how quality of life data can be utilized in cost-utility analysis: the cost per quality adjusted life year (QALY) gained by adding interferon was estimated at USD 110,000. PMID- 9417688 TI - [Significant medical quality concepts in a clinical context]. AB - For centuries, doctors have been working continuously on improving clinical practice, but there has not been the same focus on improving the organization of health care. Many doctors still believe that quality improvement only applies to medical practice. This is no longer so. Present day medical quality improvement includes both clinical practice and the health care system. The principles relating to quality systems describe the strategies and practical accomplishment of changes to improve the quality of care. These common principles should be applied to clinical practice, to how the systems work and to interpersonal relationships in health care. Norwegian medical quality assessment is encountering problems with language and definitions. A common understanding and use of standard terms is still lacking. This may be one reason why the medical profession is still unfamiliar with quality assessment as a method for the continuous improvement of health care. Using a clinical example, this article discusses the understanding and use of five important terms in quality assessment: quality, clinical guidelines, indicators, criteria, and standards. PMID- 9417689 TI - [Which medical quality concepts should be used in Norway? Some significant quality concepts with definitions, synonyms and analogues English terms]. AB - The terminology for quality in medical practice is over-complex, confusing and often applied indiscriminately. Such terms are meant to contribute to the understanding of medical performance and to be used as tools for its improvement. In order to be able to understand each other and to perform quality assessment a set of comprehensible terms and concise definitions of those terms is needed. This article defines and discusses in alphabetical order some of the most important Norwegian medical quality terms, giving the corresponding expressions in English. The aim of the authors is to create a Norwegian reference for medical quality terminology, modified to conform with international literature. This is a continuous process, as are other areas of quality assessment, and for the present time we recommend that these definitions be used as standard terminology for Norwegian medical quality assessment. PMID- 9417690 TI - [Project geriatric outpatient clinic. A study on geriatric outpatient clinics and patient basis]. AB - In 1996 PROJECT: geriatric outpatient clinic was initiated by the Ministry of Health and Social Affairs. The purpose was to collect more extensive information about geriatric outpatient clinics in Norway with regard to organization of the clinics, the types of activities and the clinics and the patients attending them. Information on the activities in 1995 was collected, and a summary prepared. Detailed records were made of the activities at 15 geriatric outpatient clinics during March 1996: 288 initial contacts and 239 repeat contacts were recorded. Assessment for mental impairment and multimorbidity constituted 41% of the referrals. The average number og patients per opening hour was rather low: 0.7 (range 0.08-2.5). The pay-back for the hospital was low and there was little incentive to operate the clinics. PMID- 9417691 TI - [Selection of patients for geriatric treatment--a quality assurance project]. AB - Numerous patients admitted to acute hospitals for emergency treatment are elderly, 75 years and above. These patients often have multiple diseases and severe functional limitations and disabilities. Trials have shown that geriatric assessment programmes carried out in hospital settings improve the prognosis of survival, and can prevent or postpone functional decline. However, correct selection of patients is necessary in order to succeed. The aim of this development project was to describe how a geriatric team could screen hospital in patients for geriatric treatment. The characteristics of 102 patients selected for geriatric treatment were compared to the criteria for a geriatric patient as defined by Laake. There was a high level of agreement between the global clinical assessment made by the geriatric team and Laake's criteria. PMID- 9417692 TI - [Rehabilitation of elderly stroke patients in a geriatric department. Course and prognosis]. AB - Stroke is an age-related disorder where nearly 70% of the patients are over 70 years of age. More knowledge about the outcome and prognosis among the eldest stroke victims is needed. We studied 171 elderly stroke patients admitted to geriatric wards for rehabilitation. The patients were assessed on admittance to and discharge from hospital, and six and 12 months after the stroke. The mean age was 78.4 years. During the first year, 19% died and 25% were admitted to nursing homes. After 12 months six out of ten patients were living at home. Our results indicate that even elderly stroke patients have a potential for functional improvement after a stroke. PMID- 9417694 TI - [Should the Norwegian health care services be reorganized to assure quality?]. PMID- 9417693 TI - [Ginseng--no identifiable effect in geriatric rehabilitation]. AB - The consumption of ginseng root has increased greatly in the Western world during the last decades. Because clinical trials have indicated a positive effect of ginseng on physical and psychomotor performance, we have undertaken a trial of Gericomplex to evaluate ginseng as an adjuvant in the treatment and rehabilitation of geriatric patients. The length of stay in hospital and activities of daily living served as the principal study variables, and cognition, somatic symptoms, depression and anxiety were also assessed. No positive effect of Gericomplex as an adjuvant in geriatric rehabilitation was seen. PMID- 9417695 TI - [Care of epilepsy in mentally retarded persons]. PMID- 9417696 TI - [Who needs to be protected from himself?]. PMID- 9417697 TI - [Fracture--just bone density?]. PMID- 9417698 TI - [Steroid treatment of asthmatic children]. PMID- 9417700 TI - [Is it asthma or chronic obstructive pulmonary disease?]. PMID- 9417701 TI - [Diagnosis of renal diseases]. PMID- 9417699 TI - [Sedation of children on respirator]. PMID- 9417702 TI - [Aseptic loosening of cemented, total hip alloplasty]. AB - The cemented low-friction arthroplasty of the hip (THR) has emerged as one of the most successful operations. About 3.500 THR's are inserted every year in Denmark, the clinical success rate at ten years being 90-95 percent. The most frequent long-term complication remains aseptic loosening of the prosthesis. This mechanism of failure is still a matter of discussion and it seems that aseptic loosening is a rather complex and multifactorial process. Our review deals with the various hypotheses with special reference to particulate debris generated at the prosthesis' surfaces and its active biological role in inducing aseptic loosening. PMID- 9417703 TI - [Sedation of children on respirators. A questionnaire study on therapeutic strategies and withdrawal problems]. AB - The purpose of this study was to identify the patterns of use of sedative agents and withdrawal problems in the paediatric intensive care unit (PICU). A questionnaire survey was mailed to 46 Danish intensive care units. Only 24 of these treated paediatric patients needing mechanical ventilation. The agents most frequently employed were morphine and midazolam. There was general satisfaction with the used drugs. The majority of departments noted that there was no protocol for sedation in their PICU. The majority of departments (70.8%) did not consider drug withdrawal to be a problem, but those departments who treated most children considered drug withdrawal to be a problem. PMID- 9417704 TI - [Treatment of severe acute exacerbation of asthma and chronic obstructive lung disease. An interview study]. AB - A telephone survey was conducted of all the 71 Danish hospitals with the capacity to receive acutely ill medical patients. The purpose was to register treatment regimes used in acute asthma and exacerbations in chronic obstructive pulmonary disease (COPD). The house officer on duty was interviewed and questioned about the use of nebulizers, oxygen therapy, bronchodilators, steroids, theophyllins and monitoring of the patient's condition. The physician survey was supplemented by a smaller survey among emergency room nurses about nebulizing systems. The answers showed inadequate knowledge of nebulizing systems. There was a noticeable variation in the dosing of oxygen and in the dosing of bronchodilators and steroids. beta 2-agonist treatment by nebulizer differed with a factor 14 in dose. The majority of the physicians had no specific parameters for monitoring severity of disease. CONCLUSION: There is a need for improvement of the knowledge of nebulizing systems, including specific knowledge of the appropriate use of propellant gasflow and time of nebulizing for optimum performance of the used nebulizer. Divergent answers from the nurses and the physicians show the need for interdisciplinary instruction. The noticeable variation in treatment in this Danish survey displays a need for quality control in terms of concise guidelines for medical therapy in acute exacerbations of asthma and COPD and guidelines for monitoring of the response to the treatment. A suggestion for a treatment regime is proposed. PMID- 9417705 TI - [Distress symptoms in hospice patients]. AB - A number of symptoms cause physical or mental distress and suffering in the terminal and dying patient. In this prospective study of 117 patients (96% with a cancer diagnosis) in a Danish hospice all symptoms causing distress were assessed daily in three degrees of severity. The ten most frequently recorded symptoms were: fatigue, pain, weakness, dyspnoea, immobility/paresis, anorexia, general malaise, nausea/vomiting, oedema and amnesia. Fatigue was registered on 60.9% of the admission days, pain on 27.3%, dyspnoea on 19.2% and nausea/vomiting on 8.5%. The prevalence of pain, dyspnoea, nausea/vomiting, thirst and anxiety did not increase during the last seven days of life. Unconsciousness occurred in 23% of the patients during the last 24 hours and in 5% on the day before. PMID- 9417706 TI - [Ambulatory knee arthroscopy in arthroscopic surgery under local anesthesia]. AB - After introducing knee arthroscopy in local anaesthesia as a standard regime in outpatients, we want to describe our experiences from 403 arthroscopies performed in 401 patients, aged 15-80 years, mean 37 years. Diagnostic arthroscopy was performed successfully in 397 of 403 cases. Arthroscopic surgery was indicated in 203 patients and carried out in 188 cases. It was possible to resect 144 of 158 tears of the meniscus. In conclusion, outpatient arthroscopy of the knee with transarthroscopic surgery in local anaesthesia using high dose lidocaine and no sedation is a valuable tool, and should be used on a larger scale. It is possible to carry out diagnostic arthroscopy and arthroscopic surgery in most patients, especially concerning resections of menisci. PMID- 9417707 TI - [Intravascular malignant lymphomatosis]. AB - Intravascular malignant lymphomatosis (IML) is a rare form of extranodal non Hodgkin lymphoma characterized by proliferation of malignant lymphoid cells within the lumen of small blood vessels. We describe a case of IML presenting with non-specific pulmonary symptoms, weight loss, intermittent fever and a confusing collection of laboratory findings. Later on the patient developed cardiac symptoms, and finally diffuse cerebral symptoms and skin lesions. His condition deteriorated and he died within six months. The diagnosis of IML was made at autopsy. Complete remission and long-term disease-free survival may be obtained with standard chemotherapy directed at high-grade lymphomas. It is important to remember IML in the differential diagnosis of patients with confusing and changing ischaemic symptoms and signs from several organs. PMID- 9417708 TI - [Perioral dermatitis in children under steroid inhalation therapy]. AB - Topically applied corticosteroid is a known provoking factor in perioral dermatitis. In recent years inhaled corticosteroids have become first line therapy for asthma in children. We present two characteristic cases of perioral dermatitis that developed during asthma treatment with budesonide using either a nebulizer or a spacer connected with a mask. In severe cases treatment with oral erythromycin or topical metronidazole gel may be considered. The eruption should not be confused with atopic eczema. PMID- 9417709 TI - [Infections--premature labor and cerebral paresis]. PMID- 9417710 TI - [Pleural plaques--international status]. PMID- 9417712 TI - [Doctor's delay]. PMID- 9417711 TI - [Campaign of the EU for olive oil...]. PMID- 9417713 TI - [Why is the incidence of breast cancer increasing?]. PMID- 9417714 TI - [Patient in the communication gap between primary and secondary health sectors]. PMID- 9417715 TI - [Diabetes mellitus and malfunctions of insulin secretion]. PMID- 9417716 TI - [Angiotensin-converting enzyme inhibition and progressing chronic nephropathy]. PMID- 9417717 TI - [Cardiovascular gene therapy]. AB - Recent insights into the pathogenesis of cardiovascular disease have created the background for development of molecular therapies, and somatic gene transfer and use of antisense oligonucleotides are among the exciting new technologies in this field of research. Gene transfer into the cardiovascular system can be achieved by non-viral or viral vectors, and the latter (i.e., recombinant retrovirus or adenovirus) have the greatest efficacy in vivo. There are several potential clinical applications of gene therapy in cardiovascular disease, and interest has focused on restenosis after angioplasty, critical limb ischaemia, venous bypass graft occlusion, myocardial infarction, and familial hypercholesterolaemia. Favourable results of gene therapy and/or antisense strategies have been reported in experimental models, and these approaches show considerable promise for the treatment of human cardiovascular disease. However, definite clinical efficacy remains to be demonstrated in any gene therapy protocol, and there are many unresolved issues before gene therapy is likely to be implemented in the cardiovascular clinical practice. PMID- 9417718 TI - [Psychopharmacological treatment of treatment-resistant schizophrenia]. AB - The research criteria of treatment-resistance in schizophrenia are reviewed and discussed, and the following definition for the use in the clinic is proposed: At least two periods of treatment with conventional antipsychotics from two different chemical classes at adequate doses for a period of at least four weeks, each without significant symptomatic relief. In addition, patients have to meet the criteria of clinical significant psychopathology, reduced psychosocial function or reduced quality of life. The article also reviews efficacy studies and the recommendations for the psychopharmacological treatment of therapy resistant schizophrenia. Placebo-controlled and open trials have shown clozapine to be clearly effective in 30-60% of patients resistant to conventional antipsychotic treatment. Up to one half of the patients may respond between three months and a year of treatment. Clozapine has been found to reduce positive and negative symptoms as well as enhancing both cognitive and psychosocial function in schizophrenics. The first drug of choice today, after treatment-resistance has been established is a trial of clozapine for at least six months. The optimal dose range of clozapine appears to be 300-600 mg/day for most patients. Some schizophrenics cannot be treated with clozapine because of contraindications or do not respond sufficiently. In these patients conventional antipsychotics have to be prescribed and augmentated with benzodiazepines, antidepressants, mood stabilizers, or electroconvulsive treatment. PMID- 9417720 TI - [Communication between primary and secondary sector and its significance for patient care. Evaluation based on admission records and discharge letters of patients admitted acutely to a department of internal medicine]. AB - The aim of this study was to evaluate the informative content of referrals and discharge summary letters concerning patients admitted acutely to the Department of Internal Medicine in a Danish district hospital, and to estimate the delivery time for discharge summary letters. A total of 132 patients were consecutively enrolled from May 2nd to May 30th 1995. The receiving doctor in the hospital on the day of admission as well as the general practitioner who received the discharge summary letter were asked to complete a specific designed questionnaire. The referrals were often insufficient regarding patient history (in particular data on psychosocial aspects and prescribed medicine) and physical examination. Every fifth referral was never received by the hospital. The delivery time for discharge summary letters was 13 days (0-87 days). In every fifth case the information was needed earlier. The discharge summary letters often lacked information concerning what the patients had been told, prognosis and suggestions concerning social medicine. This study indicates that the collaboration between general practitioners and doctors in hospital can be improved by of enhancing the informative content of referrals and discharge summary letters and the expedition time of discharge summary letters. PMID- 9417719 TI - [Incidence and mortality of breast cancer among women in Denmark 1943-1992]. AB - Nationwide incidence and mortality rates are presented for female breast cancer in Denmark from 1943 to 1992. The annual incidence rate of breast cancer has increased among Danish women from 61 in 1943-1947 to 105 per 100,000 women in 1988-1992. Mortality rates have increased to a smaller extent, from 36 to 43 per 100,000 women. We found that only a small part of the observed increase in incidence of breast cancer can be explained by the known risk factors, based on calculation of population attributable-risk per cent. PMID- 9417721 TI - [An ambulatory service staffed with military physicians in a rural district. Experiences from a period of 6 years]. AB - A local rendez-vous arrangement is described retrospectively in which the medical officer on duty at the infirmary, Oksbol military camp participated in the ambulance service and the prehospital treatment of acutely ill patients. The military ambulance supports the civilian ambulance service in the municipality of Blavandshuk, Western Jutland. This arrangement was carried out in 430 cases and 399 patients were brought to hospital. More than 14% of the services did not result in transportation. Forty-seven percent of the services were due to accidents and 30% to illness. In 16% of all cases the response-time was less than five minutes. All the patients were classified according to the Oksbol-score. Injuries were diagnosed in 48% and cardiovascular disease in 19%. Sixty-seven percent were treated immediately by the military ambulance. This arrangement has improved the prehospital treatment of acutely ill patients by using pre-existing resources from a military camp. We propose further cooperation between civilian health authorities and the Danish Armed Forces' Health Services when planning the prehospital services. PMID- 9417722 TI - [Antibiotic-induced hemorrhagic cystitis]. AB - Two cases of haemorrhagic cystitis following treatment with methicillin and penicillin G are presented. Two males, aged 24 and 45 years, presented identical symptoms including haematuria, dysuria and pollakisuria. The condition has in rare instances been described as caused by antibiotic treatment; in all cases a penicillin was involved. All symptoms promptly vanished when the antibiotic treatment was stopped, and the reactions were possibly allergic since cross reactions between different penicillins have been described in earlier cases. PMID- 9417723 TI - [Primary sclerosing cholangitis with itching treated during pregnancy with ursodeoxycholic acid]. AB - A 23-year old woman with primary sclerosing cholangitis was being treated with ursodeoxycholic acid (URSO). When pregnancy was diagnosed, she was already through the first trimester. The treatment was discontinued, but her symptoms including severe pruritus recurred immediately, and the biochemical markers of bile duct obstruction worsened. It was necessary to start URSO-treatment again, which relieved her symptoms and improved the biochemistry. The remaining part of the pregnancy was uncomplicated, there were no malformations and so far, the baby is doing fine. URSO may be a possible treatment for severe intrahepatic cholestasis and pruritus during pregnancy. PMID- 9417724 TI - [Prevention of breast cancer--simple answers on complicated questions]. PMID- 9417725 TI - [Olanzapine. A new antipsychotic agent]. PMID- 9417726 TI - [Electric accidents--ictus electricus--and ECG monitoring]. PMID- 9417727 TI - [Arterial diagnostics]. PMID- 9417728 TI - [Venous diagnostics]. PMID- 9417729 TI - [Treatment of chronic cardiac insufficiency with coenzyme Q10, results of meta analysis in controlled clinical trials]. AB - Meta-analysis applied to eight controlled clinical trials of coenzyme Q10 (CoQ10) treatment of congestive heart failure revealed a significant improvement a several important cardiac parameters such as ejection fraction (EF), stroke volume (SV), cardiac output (CO), cardiac index (CI) and end diastolic volume index (EDVI). Concerning the improvement in SV and CO the average patient in the CoQ10 group had a higher score than respectively 76% and 73% of the patients in the placebo group. The improvement in CO and SV was also significant when considering of homogeneity. Additional controlled clinical trials seem justified which may strengthen the power of the meta-analyses. However, based on available results, it can not be excluded that CoQ10 may have a future role a adjunctive therapy in a dosage of 100-200 mg/day in the treatment of chronic congestive heart failure. PMID- 9417730 TI - [Efficacy and safety of dietary supplementation containing Q10]. AB - The literature concerning the importance of Q10 for health and disease has been reviewed. Dietary intake together with normal in vivo synthesis seems to fulfil the body's demands for Q10 in younger, healthy individuals. The importance of Q10 in general well-being has not been investigated in controlled experiments. The literature allows no firm conclusions about the significance of Q10 in physical activity. In different cardiovascular diseases a positive effect of oral Q10 supplementation has been reported, especially in chronic heart failure. These effects should be further examined. No important adverse side effects have been reported from experiments using daily supplements of up to 200 mg of Q10 for six to twelve months, and 100 mg daily for up to six years. PMID- 9417731 TI - [Electric injuries--cardiac monitoring?]. AB - During the years 1980-1990, 49 patients (38 men and 11 women) were treated in the department for various electrical injuries. The average age was 27 years (2-78 years). Thirty-nine had suffered low voltage (less than 1000 volts) accidents and ten had suffered high voltage (more than 1000 volts) accidents. The incidence and type of cardiac arrest/loss of consciousness on the scene of the accident, myoglobinuria, acute fasciotomies, surgical interventions, amputations, cardiovascular complications and outcome is reported. The most important finding was that no cardiac abnormalities were seen even with the patients with a clinical primary cardiac arrest. In spite of this finding 18 of the patients were initially brought to a medical department for electrocardiographic monitoring. This could clearly delay surgical intervention, though it did not seem to affect the final outcome (with some reservations). It was concluded that only abnormal ECGs should indicate electrocardiographic monitoring. Otherwise an electrical injury is a surgical matter. PMID- 9417732 TI - [Zinc chloride smoke pollution. Effects of minimal exposure]. AB - Thirteen patients were exposed to accidental zinc chloride inhalation during an army exercise. Smoke bombs were released in open air. The exposure was modest ranging from "taking a few inhalations" to "5-10 minutes in a house with smoke drifting in through unshuttered windows". Initial symptoms were scanty. All patients received inhalation steroid on admittance followed by i.v. bolus of hydrocortisone. Four patients continued systemic steroid treatment (prednisolone 40 mg with stepwise reduction to zero over four weeks) because exposure was judged significant (> 1 minute of unprotected inhalation). No respiratory symptoms developed within an eight week observation period. However, a gradual decline in pulmonary CO diffusion capacity (to 85% (76-99 of initial capacity) was observed within the first four weeks. It is concluded that a very modest inhalation of zinc chloride smoke may induce prolonged impairment of pulmonary function. PMID- 9417733 TI - [Discrepancies between diagnosed asthma, asthma-like symptoms and a test for abnormal lung function]. AB - For the diagnosis of asthma, it is neither clear to which degree various tests and symptoms identify the same subjects nor how these characteristics are best combined. We assessed the interrelationship between physician-diagnosed asthma, asthma-like symptoms and abnormal airway function in a population based sample of 495 12-15 year old schoolchildren. Participants filled in a questionnaire and underwent baseline spirometry (FEV1%), provocation with treadmill exercise (EXE) and with inhaled methacholine (PD15), and monitoring of peak expiratory flow (PEF) twice daily for two weeks. Most symptomatic subjects with any test positive were identified by PD15 alone (75%) or in combination with PEF monitoring (89%). Although interest agreement was weak (kappa < 0.40 for all pairs), significant associations were found between PD15 and EXE, between PEF and EXE and between FEV1% and PD15. However, PEF variability and methacholine responsiveness seem to identify different varieties of airway pathophysiology, and the combined use of the two tests may be helpful as an epidemiological screening tool for asthma. PMID- 9417734 TI - [The validity of data in registration of operations. A quality analysis]. AB - In Denmark all operations are registered centrally and identified by a unique number describing the exact operation. The aim of this study has been to validate the data, which are registered. Within a period of four months all registrations were carried out in duplo in two independent registers. The contents of the two registers were compared manually. A total of 1568 operations were registered. In one of the registrations 102 (6.5%) were missing and in the other 112 (7.1%). In 70 cases (4.5%) a discrepancy was found between the registers according to type or date of operation. It is concluded, that about 10% of all registrations in such a register are wrong. A better validity of data might be obtained by a simplification of registration procedures. Double registration is an effective way to locate the mistakes. PMID- 9417735 TI - [Spondylitis and medullary transverse syndrome caused by Staphylococcus aureus]. AB - Three cases of haematogenous osteomyelitis in the vertebral column caused by Staphylococcus aureus are reported. The cases, which were associated with severe neurological symptoms and/or death, were initially characterized by a long period with no or discrete local signs of infection and by values of temperature and leucocyte counts within or close to normal values. In this period measurements of the sedimentation reaction and C-reactive protein were elevated, and were markers of persistent infection. At the Department of Clinical Microbiology in the county of Copenhagen blood cultures from a total of 49 patients were found to be positive for S. aureus during the period January to March 1996. Six patients were found to have osteomyelitis (12%, including four cases of spondylitis) and nine patients were suspected of having osteomyelitis. This frequency of patients with S. aureus bacteraemia having osteomyelitis was significantly higher than reported in another Danish study (10), which together with the severe outcome of the infection emphasizes the need for attentiveness to these serious complications of S. aureus bacteraemia. PMID- 9417736 TI - [Prenatal diagnosis]. PMID- 9417737 TI - [Overtreatment at emergency departments?]. PMID- 9417739 TI - [Development of the "natural healing" concept]. PMID- 9417738 TI - [Intracranial lipomas]. PMID- 9417740 TI - [Natural healing methods versus alternative medicine--attempt at a distinction]. AB - In this decade we can realize--especially in Europe--that many patients turn their back on established medicine and want to undergo alternative medicine. Alternative medicine is often mixed up with natural medicine, although classical natural medicine is very different from alternative medicine and is a part of established medicine. This paper gives a survey of the important methods of natural medicine on one side and of alternative medicine on the other side including their difference. PMID- 9417741 TI - [Unconventional treatment methods in multiple sclerosis and their expert assessment]. AB - Unorthodox therapies are used also by MS-patients very frequently. Not all procedures are harmless affecting only the purse. The medical expert working within the German health care system will also be called for questions of compensation of costs. A survey of some paradigmatic unorthodox treatments is given. While the law and the jurisdiction concerning the public health insurance fund pronounce clear standards in respect to the demands on the efficacy shown by modern (biomathematic) methods, it is essentially a more difficult task to bring the jurisdiction of the Federal Supreme Court (BGH) in harmony with basic requirements of a rational and scientific medicine. The interindividual and intraindividual course of MS is most variable, therefore it is very difficult to make a distinction between natural course and effects of a therapy in a single case while the only possibility to estimate individual therapeutic chance is a statistic evidence of efficacy in general. PMID- 9417742 TI - ["Control of symbiosis" or "microbiological therapy"--an immunological therapy principle?]. AB - "Flora modulation" or "microbiologic therapy"-- an immunologic method of treatment? "Microbiologic therapy" includes the methods of quantitative measurement of the intestinal "microbial flora", the oral or parenteral application of microbial pharmaceutics and the preparation of autovaccines from excretions of the treated persons. Initially a "flora modulation" was supposed to be the mechanism of "microbiologic therapy". After the failure of this hypothesis, some physicians claim the "microbiologic therapy" to be a special form of immunomodulation or -stimulation. Most influential in this immunologic foundation of "microbiologic therapy" was the "Institut fur Mikrookologie" in Herborn, Germany. A detailed analysis of the available publications however reveals, that all methods of "microbiological therapy" are based on theoretically untenable presumptions. Furthermore, up to now there is no scientific evidence for the effectiveness of this form of therapy. PMID- 9417743 TI - [Assessment of work capacity and occupational rehabilitation in curatively treated tumor patients]. AB - Cure and long survival of cancer patients are often accompanied by reductions of performance and vocational handicaps due to the treatment or due to the cancer itself. These reductions vary according to the kind of cancer, the different therapies and the different doses as well. There is a considerable risk of late effects of chemo- and radiotherapy that can be evaluated more precisely today than in the past. Rehabilitation is able to reduce or even to prevent or compensate these reductions of performance and vocational handicaps. Rehabilitation basing on roborating measures is not suitable for these possibilities and should be given up to the benefit of a more flexible rehabilitation of cancer patients being determined by the extent of impairment and functional deficiency. This rehabilitation work includes the consideration of vocational handicaps as well. PMID- 9417744 TI - [Inpatient treatment of a patient with large duodenal diverticulum and psychogenic vomiting--somatic and psychosomatic approach]. AB - The inpatient psychotherapy of a female patient with a large duodenal divertivulum and psychogenic vomiting is described. The specifics of the inpatient treatment setting necessary for change are presented thereby demonstrating to be helpful for these kind of patients with previous negative experiences of somatic and psychotherapeutic treatment. PMID- 9417745 TI - [Promoting motivation in acute inpatient treatment of alcohol dependent patients]. AB - The necessary conditions of a motivating withdrawal of alcohol-addicted inpatients based on the experiences with drug addicts are pointed out. If inpatient treatment is necessary either a more complex treatment of qualified withdrawal (QW) or a short-term intervention to manage with the crisis (CI) is offered to the addict according to his actual motivation. In the CI-programme (39 cases) 41% of the patients were able to switch over to the QW-programme. The other patients were dismissed quickly (median period of treatment 8 days). In the QW-programme (88 cases) 70% of the patients finished the treatment regularly and subsequent treatment was successfully gained (23% in-patient rehabilitation, 7% in-patient psychotherapy and 12% outpatient therapy). PMID- 9417746 TI - [10 years radiofrequency ablation of accessory conduction pathways]. AB - Catheter ablation of the preexcitation syndrome is a curative treatment. Accessory pathways between structures of the specific conduction system or working myocardium are morphologic prerequisites for orthodromic or antidromic reentrant tachycardia of various frequency, duration, and rate. Detection of mechanisms of tachyarrhythmias and understanding the role of accessory pathways in tachycardia, mapping of accessory connections, function of nodal conduction system, and additional accessory pathways are necessary for successful catheter ablation. Anomalies of the tricuspid valve and coronary sinus and concomitant disease of the heart should be investigated prior to catheter ablation using echocardiography and contrast injection. Different variations of the preexcitation syndrome and the results of catheter ablation in 300 patients are demonstrated. Catheter ablation is indicated in refractory tachyarrhythmias on the basis of accessory pathways. The treatment is performed in a catheterization laboratory by two highly experienced cardiologists in the field of electrophysiology after training in 100 procedures. The investigators need experience in interventional treatment of coronary artery disease, in transseptal puncture, and in the management of complications (coronary and valvular problems, thromboemboly, and pericardial drainage). Catheterization needs careful protection of radiation. An ablation is possible at an atrial or ventricular insertion site of the accessory pathway or in between. Ablation is done during sinus rhythm, atrial stimulation, antidromic reentry, or atrial fibrillation or during ventricular pacing and orthodromic tachycardia. The procedure should end with bidirectional block of the accessory pathway in 90-95% of the patients. Complications occur in 2-4% of procedures. Recovery of accessory conduction is observed in 8%. Catheter ablation of the accessory pathway is the treatment of first choice in symptomatic patients with the pre-excitation syndrome. The procedure has limited risks and a high success rate. PMID- 9417747 TI - [Decreased apoptosis as a pathogenic factor in intimal hyperplasia of human arteriosclerosis lesions]. AB - Restenosis remains a persistent problem following intravascular reconstruction. Smooth muscle cell proliferation, extracellular matrix production and remodeling are accepted mechanisms of restenotic lesion formation. Decreased programmed cell death (apoptosis) may also contribute to restenosis by prolonging the life span of intimal cells, with their subsequent accumulation and development of hyperplastic lesions. The objectives of the present study were as follows: i) to identify cell death, ii) to distinguish and quantify apoptosis from necrosis, and iii) to compare restenotic with primary lesions. To this end, human atherectomy specimens from 25 primary and 14 restenotic coronary and peripheral lesions were studied by TUNEL test (TdT-mediated dUTP Nick End Labeling; detection of cell death by the presence of fragmented DNA), transmission electron microscopy and morphometric analysis. Intimal hyperplasia was more consistent with restenosis than with primary lesion origin, and was mainly attributed to increased smooth muscle cell density (649 vs. 219 cells/mm2; p < 0.001). The main finding of the present study is that hypercellular restenotic tissue contains fewer TUNEL+ cells than hypocellular plaques (14% vs. 27%; p < 0.05). Most importantly, ultrastructural evaluation revealed a markedly reduced portion of intimal plaque cells, especially smooth muscle cells exhibiting distinct morphologic signs of apoptosis (3% vs. 13%; p < 0.001). In contrast, incidence of necroses did not differ between both lesion types (0.13 vs. 0.12 necroses/ cell; p = 0.49). Thus, our data indicate apoptosis and not necrosis to be the crucial cell death form to account for the apparent discrepancy found in both lesion types with reduced apoptosis in cell-rich restenoses. The findings of the present study suggest that decreased apoptosis is an important regulatory mechanism ultimately leading to intimal hyperplasia as commonly found in human restenosis post angioplasty. PMID- 9417748 TI - [Low molecular weight heparin, reviparin, after PTCA: results of a randomized double-blind, standard heparin and placebo controlled multicenter study (REDUCE Study]. AB - BACKGROUND: Unfractionated heparin and its low molecular fragments possess antiproliferative effects and have been shown to reduce neointimal smooth muscle cell migration and proliferation in response to vascular injury in experimental studies. OBJECTIVES: The specific objective of the REDUCE trial was to evaluate the effect of a low molecular weight heparin on the incidence and occurrence of restenosis in patients undergoing percutaneous transluminal coronary angioplasty. METHODS: The REDUCE trial is an international prospective, randomized, double blind, multicenter study. Twenty-six centers in Europe and Canada enrolled 625 patients with single lesion coronary artery obstructions suitable for PTCA. Three hundred and six patients received reviparin as a 7000 U bolus before PTCA followed by 10,500 U as an infusion over 24 hours and then twice a day 3500 U s.c. application for 28 days. The 306 patients in the control group received a bolus of 10,000 U unfractionated heparin followed by an infusion of 24,000 U over 24 hours. These patients then received 28 days of s.c. placebo injections. The primary endpoints were efficacy (defined as a reduction in the incidence of major adverse events, i.e., death, myocardial infarction, need for reintervention or bypass surgery), absolute loss of minimal luminal diameter, and incidence of restenosis during the observation period of 30 weeks after PTCA. RESULTS: Using the intention-to-treat analysis for all patients, 102 patients (33.3%) of the reviparin group and 98 patients (32%) of the control group have reached a primary clinical endpoint (relative risk = 0.98; 95% confidence limit, 0.88-1.09; p = 0.707). Likewise, no difference in late loss of minimal luminal diameter was evident for either group. Acute events within 24 hrs occurred in 3.9% of the reviparin group and in 8.2% of the control group (relative risk = 0.49; 95% confidence limit, 0.26-0.92; p = 0.027) during or immediately after the initial procedure. In the control group, 8 major bleedings occurred, and in the reviparin group, 7 major bleeding complications were observed within 35 days after PTCA. CONCLUSIONS: Reviparin use during and after coronary angioplasty did not reduce the occurrence of major clinical events or the incidence of angiographic restenosis over 30 weeks. PMID- 9417750 TI - [Malignant primary tumors of the heart]. AB - Primary tumors of the heart are rare, and a quarter of these tumors prove to be histologically malignant. Benign and malignant primary as well as secondary heart tumors belong to the differential diagnosis when a new heart murmur, heart rhythm disturbances, or cardiac insufficiency appear. In the diagnostics of intracardiac space-occupying lesions, two-dimensional echocardiography represents the method of choice, supplemental examination by computer tomography or nuclear magnetic resonance imaging may be helpful. PMID- 9417749 TI - ["Myxoma syndrome"--a "benign" disease with "malignant" course]. AB - We are reporting on a 36 year-old woman who presented with recurrent cardiac myxomas over a period of nine years. Two of the tumors typically originated in the left atrium and one in the right atrium. Tumor embolization was the presenting symptom twice, leading to reversible cerebral ischemia and minor pulmonary embolism, respectively. The third tumor remained asymptomatic and was detected during routine echocardiographic examination. Based on a positive family history of cardiac tumors, a facially pronounced hyperpigmentation of the skin and the presence of a thyroid adenoma, the diagnosis of a "myxoma syndrome" was established. Patients with "myxoma syndrome" are generally younger than their counterparts with "sporadic myxoma" (mean age at diagnosis 25 vs. 56 years) and have a high frequency of unusual skin freckling (68%). Familial clustering of cardiac myxomas is also frequent (25%). The tumors may be located in any of the cardiac chambers (87% in the atrias, 13% in the ventricles, 50% at multiple sites simultaneously) and have relatively high (18%) 5-year recurrence rate after surgical excision. Since the clinical signs of cardiac tumors are non-specific, diagnosis essentially relies on cardiac imaging by echocardiography, computer tomography, or angiography. The superiority of transesophageal echocardiography is emphasized in this report. PMID- 9417751 TI - [Clinical value of Doppler ultrasound controlled puncture of the inguinal vessels with the "Smart Needle" within the scope of heart catheter examination]. AB - Due to the increasing number of diagnostic heart catheterizations, especially in elderly patients, as well as the increase of percutaneous transluminal coronary angioplasties, we are confronted with a rise in peripheral complications evolving from difficulties in the procedure of the puncture of the femoral artery or vene. The development of greater hematomas in the area of the puncture, the formation of arterio-venous fistulas and aneurysma spuria are the foremost complications. It was the aim of the study to investigate in as far an improved puncture technique could reduce the rate of peripheral complications. In this comparative study the vessel punctures were carried out by the conventional Judkins puncture technique and a new method using a special puncture needle. (Smart Needle" R), with an integrated ultrasonic sound device. In this study 114 patients--age 23 to 82 years--undergoing heart catheterization (91 diagnostics, 23 PTCAs) were examined. In all cases a puncture of the arterial and venous femoral vessel was done. 50 patients received a puncture via "Smart Needle" and the remaining 64 patients were punctured conventionally. In contrast to the group of patients receiving conventional puncture of the femoral artery, where in 72% more than one try was needed, in all patients of the "Smart Needle" group the first puncture was successive. Concerning the puncture of the venous vessels no significant difference between the two groups was observed. This difference between arterial and venous puncture outcome results from the difference between arterial and venous flow signals detected by the ultrasonic sound device. As to the bleeding complications hematomas following to "Smart Needle" puncture occurred less frequently-in 25%--and were significantly smaller than in the conventional group where hematomas were seen in 46%. The number of patients with hematomas with diameters of more than 5 cm was twice as high in the conventionaly punctured group (28%) than in the "Smart Needle" group (14%). However, patients suffering from arterial hypertension or hemostatic disorders showed an increased risk of vascular complications. In regard to the cost-benefit relation an indication for the use of the new technique is to be seen especially in overweight patients and patients suffering to aortic stenosis or cardiogenic shock. PMID- 9417752 TI - [Quantitative effect of a standardized pressure bandage on arterial and venous circulation in the lower extremities after heart catheter examination]. AB - AIM: The aim of the study was to evaluate the effect of a conventional pressure dressing on arterial and venous blood flow of the legs after cardiac catheterization. METHODS: Duplexsonographic measurements were performed of both legs in 100 consecutive patients before catheterization and with a pressure dressing after the procedure. The pressure dressing was applied by means of a pressure pad on the punctured leg, so that the ankle-arm-index (blood pressure at the leg/blood pressure at the arm) was not influenced. Arterial and venous blood flow of the superficial femoral artery and vein were registered. We evaluated both legs by means of duplex sonography to detect arterial or venous complications after cardiac catheterization. Statistical evaluation was performed using the 1-sample Wilcoxon test. RESULTS: There was a significant reduction of venous blood flow in the punctured leg from a mean of 119 ml/min before puncture to 84 ml/min during pressure dressing (29% flow reduction, p < 0.01), and a reduction of arterial blood flow from a mean of 132 ml/min before puncture to 84 ml/min during pressure dressing (36% flow reduction, p < 0.01). In the contralateral leg, venous blood flow was reduced from about 118 ml/min to 96 ml/min during pressure dressing (19% flow reduction, p < 0.01), and arterial blood flow was reduced from about 129 ml/min to 93 ml/min during pressure dressing (28% flow reduction, p < 0.01). There were no venous complications, 5 patients suffered false aneurysm (5%), and one patient had an arteriovenous fistula (1%). CONCLUSION: A conventional pressure dressing caused a significant reduction of arterial as well as venous blood flow of both the punctured leg and the contralateral leg. This is of clinical relevance especially in patients with known peripheral arterial disease or patients at risk for deep venous thrombosis. PMID- 9417753 TI - [Histologic specimen collection in pulmonary hypertension using percutaneous, transvascular biopsy--animal experiment and initial clinical data]. AB - Pulmonary hypertension is a severe multietiological disease largely resistant to treatment. Biopsy is often necessary to establish differential diagnosis. In pulmonary hypertension transbronchial pulmonary biopsy is contraindicated and open lung biopsy is the method of choice. In our paper we report about experimental results in animals and first clinical data on a new biopsy technique (percutaneous transluminal biopsy, PTB) providing access to histology in pulmonary hypertension without the need of surgery. In a subset of animals pulmonary hypertension was produced experimentally. In addition, we carried out the PTB in patients with bronchial carcinoma. Our preliminary results demonstrate that sufficient material could be gained for the histologic evaluation of pulmonary parenchyma and in pulmonary vessels. No complications have been noted so far. PMID- 9417755 TI - [Dextrocardia and Poland syndrome in a 59-year-old patient]. AB - Poland syndrome consists of unilateral absence of the sternal head of the pectoralis major muscle, ipsilateral symbrachydaktylia, and occasionally associated other malformations of the anterior chest wall, mammilla, and mamma. To our knowledge a simultaneous occurrence with dextrocardia was reported in seven patients. We report on an additional patient with this unusual coincidence. A 59-year old man showed mild symbrachydaktylia of the left hand and deformity of the left part of the anterior chest wall, both present since birth. We found an additional dextrocardia with situs solitus, d-loop, and correctly connected great arteries. Besides arterial hypertension there was no remarkable impairment of the patient's condition. PMID- 9417754 TI - [Acute myocardial infarct caused by a muscle bridge of the anterior interventricular ramus: complicated course with vascular perforation after stent implantation]. AB - A 47-year-old male patient was admitted to our hospital with acute anterior myocardial infarction. Immediate coronary angiography was carried out, which showed proximal occlusion of the left anterior descending artery (LAD). After mechanical recanalization, a reduction in vessel caliber at the site of occlusion was visible, and balloon angioplasty with consecutive stent implantation because of vessel wall dissection was performed. After the procedure, diameter reduction of the entire vessel segment distal to the stent and muscular bridging with subtotal systolic obliteration of the LAD and one diagonal branch were demonstrated. Diastolic coronary flow did not appear to be limited (TIMI 3). Dipyridamole-thallium cardiac imaging revealed an incomplete perfusion defect of the anteroseptal region and a reversible perfusion reduction of the anterolateral region. For definitive treatment, we decided to implant a 3.0 mm-stent at the site of muscular bridging. Although balloon sizing was adapted to the diameter of the proximal reference segment, measured by quantitative coronary angiography, coronary perforation into the right ventricular outflow tract due to balloon oversizing in the distal dilation segment occurred. The patient remained asymptomatic at rest as well as under exercise testing, and hemodynamics remained stable. Coronary re-angiography after 1 week demonstrated a persistent fistula with complete opacification of the LAD and normal coronary flow (TIMI 3). Within the following 3 months, the coronary fistula closed spontaneously. CONCLUSIONS: Muscular bridging is a rare cause of acute myocardial infarction. Balloon angioplasty and stent implantation in the bridged segment may be complicated by coronary artery perforation due to balloon oversizing. Risks and benefits of this therapeutic option, therefore, have to be critically evaluated, and careful selection of balloon size using measurements of proximal and distal reference diameter assessed by intravascular ultrasound is recommended. Coronary artery perforation into the myocardium with subsequent development of a fistula may be treated conservatively as long as the patient remains asymptomatic. The frequency of spontaneous closure of the fistula is high. PMID- 9417756 TI - [Re: U. K. H. Wiegand, R. Schneider, F. Bode, A. Brandes, G. Taubert, J. Potratz (Lubeck/Ludwigshafen). Atrial detection of AV synchronicity in "single-lead" VDD pacemakers. Is the occurrence of atrial undersensing predictable?]. PMID- 9417757 TI - [Hematopoietic stem cell transplantations in treatment of autoimmune diseases]. AB - Autologous and allogeneic blood or marrow transplants are currently under intensive discussion as therapeutic option for patients with severe autoimmune disorders. Preclinical data as well as case reports from patients with transplants for other indications and concommitant autoimmune disorders support such concepts. In order to advance in a coordinated way the European League against Rheumatism EULAR and the European Group for Blood and Marrow Transplantation EBMT prepared common guide lines on how to proceed. They are presented and summarised. Clear clinical data are still missing. Preliminary data are promising and protocols for prospective studies are being prepared. They should allow a better evaluation of the procedures in a few years from now. PMID- 9417758 TI - [Cyclophosphamide therapy in systemic lupus erythematosus]. AB - Cyclophosphamide is a potent immunosuppressive drug which is widely used to treat renal and central nervous system manifestation of Systemic Lupus Erythematosus (SLE). It is employed especially to prevent renal failure in lupus nephritis. Within the last decade intravenous cyclophosphamide bolus therapy has become the standard therapy in severe SLE due to its tendency towards a higher efficacy and fewer side effects as compared to oral application. Studies on the slightly more cost-effective oral cyclophosphamide bolus therapy have recently been published, however, there are no data on long-term results yet. Here, we review the effects and side effects of cyclophosphamide as well as recent clinical studies on cyclophosphamide bolus therapy in SLE. Because of the possible side effects, especially the high risk of malignancies, which sharply increases at a cumulative dose of approximately 60 g, cyclophosphamide should be used cautiously. Cyclophosphamide bolus therapy should only be performed in hospitals with special experience. PMID- 9417759 TI - [Arthroscopic synovectomy of the knee joint in rheumatoid arthritis]. AB - The role of arthroscopic synovectomy in Rheumatoid arthritis is still in discussion. The evaluation of the available scientific data out of the literature should give a contribution to this discussion. The review of the studies done in only small numbers of patients demonstrates good clinical results for a short or middle lasting follow-up time. The evaluation showed a lot of differences and problems in the study design, amount of usable datas and results. For the scientific rating of the results of arthroscopic synovectomies it is necessary to have prospective studies with sufficient and reliable data on activity of the disease, the pre-, post- and intraoperative findings, standardized surgical procedures and longer lasting follow-up times. PMID- 9417760 TI - [Follow-up studies after bicondylar superficial replacement of rheumatoid knee joint destruction]. AB - This study documents prospectively the Knee Society knee- and function score of 28 patients with rheumatoid arthritis with 34 PFC unconstrained total knee arthroplasties from preoperative values on at yearly intervals. The average follow-up period was 3.4 years (range 2-5.5 y). At last follow-up over 80% of the knees were painfree. All but one patient could walk more than 500 m. Knee and function score increased significantly from 30.1 resp. 35.0 to 83.8 resp. 74.6 (P < 0.000001). Postoperatively the knee score rose soon to a constant level whereas the function score showed a continuous slow increase up to 5 years. We observed one deep venous thrombosis and one subluxation. At an intermediate follow-up rheumatoid knees are clinically and functionally successfully operated on using an unconstrained TKA. Pain relief is excellent. We recommend the use of a scoring system assessing knee and functional results separately. PMID- 9417761 TI - [Chronic polyarthritis and radiosynoviorthesis: a prospective, controlled study of injection therapy with erbium 169 and rhenium 186]. AB - The aim of this prospective study was to evaluate the efficiency of radiation synovectomy with rhenium-186 or erbium-169 in rheumatoid arthritis. In the control groups articulosynovitis was treated by injection of triamcinolonhexacetonid. Follow-up time was 3 years. Patients of the study had to fulfill the following criteria: rheumatoid arthritis (ARA criteria 1988), patient age above 40 years, standardized medical treatment with methotrexate, (started at least 6 month prior to injection therapy, given for the entiry study time), prednisolon < or = 7.5 mg/d and diclofenac < or = 150 mg/ d, and no previous surgery or injection therapy on this/these joint/s. Shoulder, elbow, wrist, and ankle joints were treated in group 1 by injection of rhenium-186 combined with triamcinolonhexacetonid, in group 2 by injection of triamcinolonhexacetonid alone. Each treatment group included 50 joints. Digital joints underwent injection of erbium-169 combined with triamcinolonhexacetonid (group 3 = 131 joints) or triamcinolonhexacetonid (group 4 = 86) alone. During the follow up period, the joints were assessed for pain, synovial swelling, joint motion, and stage of radiological destruction (Larsen-Dale-Eek). After 3 years follow-up, 228 joints met the above named criteria: group 1 = 41 joints, group 2 = 21 joints, group 3 = 131 joints, group 4 = 53 joints. Significantly better clinical results were achieved with the combined injection of rhenium-186 or erbium-169 and triamcinolonhexacetonid. Results for PIP joints were worse than for other joints, which is explained by better immobilization of the latter ones after injection. The progression in radiological joint destruction according to the stages of Larsen-Dale-Eek during the follow-up time of 3 years (= stage at 3 years minus stage prior to treatment) corresponded to the clinical results and was significantly slower in groups 1 and 3: group 1 = 0.62; group 2 = 1.7; group 3 = 0.75; group 4 = 1.43 Therefore, we recommend radiosynovectomy with erbium-169 or rhenium-168 in combination with triamcinolonhexacetonid and consequent immobilization after injection. PMID- 9417763 TI - Proceedings of the 4th International Conference on Eicosanoids and Other Bioactive Lipids in Cancer, Inflammation, and Radiation Injury. Hong Kong, October 4-7, 1995. PMID- 9417762 TI - [Diffuse fasciitis after Borrelia infection--a case report]. AB - Diffuse fasciitis (DF) [diffuse fasciitis with eosinophilia-Shulman's syndrome] has occasionally been linked to a precedent infection with Borrelia burgdorferi. Here, we report on another case of DF in a 25 year old male, in whom Borrelia burgdorferi infection as possible inciting agent could be identified based on the patient's history and laboratory data. Efforts to microscopically demonstrate spirochetes or to amplify Borrelia-DNA by nested PCR in lesional tissue failed after antibiotic treatment had already been initiated. Although only a few cases of Borrelia associated diffuse fasciitis have been reported in the literature, the link between typical signs and symptoms as well as laboratory findings of Borrelia infection and the onset of diffuse fasciitis, starting at the primary site of EM, provide indirect evidence for a causative role of Borrelia burgdorferi as a potential infectious agent for DF. PMID- 9417764 TI - Proceedings of the workshop on individual fatty acids and cancer. Washington, D.C., June 4-5, 1996. PMID- 9417765 TI - Update in Nephrology '97: Hypertension, Lipids, and Uremia Therapy. Symposium proceedings part 1. Brooklyn, New York, May 9, 1997. PMID- 9417766 TI - Human biomonitoring of arsenic and antimony in case of an elevated geogenic exposure. AB - Part of the northern Palatinate region in Germany is characterized by elevated levels of arsenic and antimony in the soil due to the presence of ore sources and former mining activities. In a biomonitoring study, 218 residents were investigated for a putative increased intake of these elements. Seventy-six nonexposed subjects in a rural region in south lower Saxony were chosen as the reference group. Urine and scalp hair samples were obtained as surrogates to determine the internal exposures to arsenic and antimony. The analyses were performed using graphite furnace atomic absorption spectrometry except for arsenic in urine, which was determined by the hydride technique. This method does not detect organoarsenicals from seafood, which are not toxicologically relevant. In the northern Palatinate subjects, slightly elevated arsenic contents in urine and scalp hair (presumably not hazardous) could be correlated with an increased arsenic content in the soil. On the other hand, the results did not show a correlation between the antimony contents in the soil of the housing area and those in urine and hair. Except for antimony in scalp hair, age tended to be associated with internal exposures to arsenic and antimony in both study groups. Consumption of seafood had a slight impact on the level of urinary arsenic, which is indicative of the presence of low quantities of inorganic arsenicals and dimethylarsinic acid in seafood. The arsenic and antimony contents in scalp hair were positively correlated with the 24-hr arsenic excretion in urine. However, antimony in scalp hair was not correlated with seafood consumption as was arsenic in scalp hair and in urine. This indicated the existence of unidentified common pathways of exposure contributing to the alimentary body burden. Short time peaks in the 24-hr excretion of arsenic in urine, which could not be assigned to a high consumption of seafood, were detected for six study participants. This suggests that additional factors relevant in the exposure to arsenic are still unidentified. PMID- 9417767 TI - Effects of chronic exposure to low doses of trichloroethylene on steroid hormone and insulin levels in normal men. AB - The aim of this study was to examine the serum levels of insulin and some adrenal steroid hormones in men chronically exposed to low doses of trichloroethylene (TCE). A total of 85 workers participated in this study. Each worker had urine collected and analyzed for trichloroacetic acids (UTCA) on the same day that a blood sample was taken for analyses of serum testosterone, sex hormone-binding globulin (SHBG), androstenedione, cortisol, aldosterone, and insulin. The mean concentration of environmental TCE was 29.6 ppm and the mean UTCA was 22.4 mg/g creatinine (range 0.8-136.4). TCE exposure did not cause any significant changes to the adrenal steroid hormone productions. The results showed that UTCA was significantly correlated to serum insulin levels. Insulin and SHBG responded in tandem, with the highest levels found in workers exposed to TCE for less than 2 years; levels of both parameters were significantly lowered in those exposed for more than 2 years. A triphasic response in insulin levels to TCE, which depended on the duration of exposure, was noted. Initial exposure caused an acute rise in insulin levels. This was followed by a fall to normal levels in those exposed 2-4 years and then a slight rise in those exposed for more than 6 years. The mechanism for this pattern of response to TCE exposure is yet unknown. PMID- 9417768 TI - Accumulation of chlorpyrifos on residential surfaces and toys accessible to children. AB - Quantitative examination of major pathways and routes of exposure to pesticides is essential for determining human risk. The current study was conducted in two apartments and examines the accumulation of the pesticide chlorpyrifos in childrens' toys after the time suggested for reentry after application. It has been established for the first time that a semivolatile pesticide will accumulate on and in toys and other sorbant surfaces in a home via a two-phase physical process that continues for at least 2 weeks postapplication. A summation of the above for a 3-6-year-old child yielded an estimated nondietary total dose of 208 microg/kg/day. Potential exposure from the inhalation pathway was negligible, while dermal and nondietary oral doses from playing with toys contributed to 39 and 61% of the total dose, respectively. If children with high frequency mouthing behavior are considered as candidates for acute exposure to chlorpyrifos residues, the estimated acute dose could be as high as 356 microg/kg/day. Routine reapplication of pesticides could lead to continued accumulation in toys and other sorbant surfaces, e.g., pillows, with large sorbant reservoirs, which can become a long-term source of exposure to a child. Estimates of a child's nondietary exposure to chlorpyrifos associated with toys and other sorbant surfaces for a period of 1 week following application appear to be of public health concern, and studies of actual childhood exposure from this pathway are warranted in the home environment. The above information should be used to determine if current procedures for postapplication reentry are sufficient and to evaluate the need for procedures to store frequently used household toys, pillows, and other sorbant objects during insecticidal application. PMID- 9417770 TI - Relative potency of xenobiotic estrogens in an acute in vivo mammalian assay. AB - The in vivo effects of xenoestrogens are of interest in relation to their potential health risks and/or beneficial effects on humans and animals. However, the apparent in vivo potency of the examined response can be confounded by a short half-life, and the metabolism of estrogens is very dependent on the nature of conversion and/or inactivation. To minimize such variables, we examined the estrogenic potency of a range of xenoestrogens in an acute in vivo assay--the stimulation of increased uterine vascular permeability in ovariectomized mice 4 hr after subcutaneous administration. While estradiol (E 2 ) and estriol (E 3 ; a relatively weak natural estrogen) readily induced vascular responses [median effective dose (ED 50 ) <10 -9 mol], much higher amounts of xenoestrogens were required. Bisphenol A was about 10,000-fold less potent than E 2 and E 3 , and octylphenol and nonylphenol were about 100,000-fold less potent; dioctyl phthalate, benzyl butyl phthalate, dibutyl phthalate, and trichlorinated biphenol produced no effect. Coumestrol was the most active phytoestrogen, with an ED 50 between 10 -6 and 10 -7 mol; genistein was about 10-fold less potent than coumestrol, and neither daidzein nor formononetin produced any marked effect, even at doses up to 10 -5 mol. All increases in vascular permeability could be blocked by the pure antiestrogen ICI 182,780. There was no evidence that any of the compounds could act as an antiestrogen in this assay or that they could exert synergistic effects in combination. These results indicate that even short-term exposure to most of the xenobiotic estrogens can induce typical estrogenic effects in vivo , but their estrogenic potency is very weak even when assessed in an acute response. PMID- 9417771 TI - Common commercial cosmetic products induce arthritis in the DA rat. AB - Many different agents, including mineral oil and silicone, have the capacity to act as immunological adjuvants, i.e., they can contribute to the activation of the immune system. Some adjuvants, including mineral oil, are known to induce arthritis in certain strains of rats after intradermal injection or percutaneous application. The aim of this study was to determine if common commercial cosmetic products containing mineral oil could induce arthritis in the highly susceptible DA (Dark Agouti) rat. Intradermal injection of five out of eight assayed cosmetic products without further additives resulted in arthritis with synovitis. One of the products induced a very aggressive arthritis, which had declined after 5-9 weeks. When this product was also assayed for arthritogenicity upon percutaneous administration, it induced a mild and transient arthritis in 5 out of 10 DA rats, whereas control animals showed no clinical signs of joint involvement. No arthritic reaction was seen in rats after peroral feeding with the most arthritogenic product or by intravaginal application of Freund's adjuvants. Silicone gel implants in DA rats did not cause arthritis. We conclude that mineral oils included in common commercially available products retain their adjuvant properties and are arthritogenic in the presently investigated arthritis prone rat strain. There is yet no evidence that mineral oils present in cosmetics may contribute to arthritis in humans, but we suggest that this question should be subject to further investigation. PMID- 9417769 TI - Bone lead as a biological marker in epidemiologic studies of chronic toxicity: conceptual paradigms. AB - The skeleton contains the majority of the body's lead burden in both children and adults. The half-life of lead in bone is in the range of years to decades, depending on bone type, metabolic state, and subject age, among other things. Measurement of skeletal lead has benefited greatly from the recent development of X-ray fluorescence (XRF) instruments that can make rapid, safe, accurate, and relatively precise measurements of lead in bone. Two types of XRF technologies exist, LXRF and KXRF; this paper focuses on KXRF, which has been the most widely validated and used. KXRF is proving to be a powerful analytical methodology for evaluating bone lead levels as a measure of time-integrated (i.e., cumulative) lead dose in epidemiologic studies of the effects of chronic lead exposure. However, insufficient attention has been given to conceptualizing the paradigms by which bone lead levels reflect lead exposure and by which the skeleton serves as an endogenous source of lead. Consideration of these paradigms, which rely on bone lead kinetics, is necessary for the proper development of a priori hypotheses involving bone lead accumulation and release, the selection of bone sites for measurement by KXRF, and the design of epidemiologic studies involving bone lead dynamics. We discuss and present supporting evidence for a conceptual model that distinguishes two major paradigms of skeletal lead, including 1) bone lead as an indicator of cumulative lead exposure (bone lead as repository), and 2) bone lead as a source of body lead burden that is mobilizable into the circulation (bone lead as source). These two roles are not mutually exclusive. Instead, they are components of the processes controlling lead accumulation into and release from bone over time. Developing successful strategies for distinguishing these two processes in epidemiologic studies will require separate measurements of lead in cortical and trabecular bone and additional measurement of specific markers of bone mineral turnover and resorption. It may also involve developing accurate methods for evaluating lead in labile compartments of the circulation, such as plasma, as a potentially useful and responsive measure of bone lead release, of the partitioning of circulatory lead, and of the toxicological significance of lead released from bone to other target organs. PMID- 9417773 TI - Approaches to DNA mutagenesis: an overview. AB - In the last several years, the use of double-stranded DNA templates together with thermostable-polymerase PCR has essentially replaced the use single-stranded DNA templates using the thermolabile polymerase for in vitro mutagenesis. Numerous PCR methods are now available, such as overlap-extension PCR, megaprimer PCR, and inverse PCR. All these PCR methods are reliable, effective, and convenient, although they are more prone to high rates of spontaneous error in mutant DNAs than are methods using thermolabile polymerases. Some improvements, such as the introduction of methylated templates, have been employed to minimize PCR errors. On the other hand, because of the introduction of many selection measures (e.g., restoration of antibiotic resistance, restoration of replication origin and unique site elimination), both double-stranded and single-stranded DNAs can now be used as templates for mutagenesis using thermolabile polymerase methods. For PCR methods, selection measures such as nested PCR has developed. All these selection measures have greatly improved the efficiency of mutagenesis by removing wild-type templates prior to transformation. Many efficient methods are available for both SDM and REM. Mutations can be introduce in vitro or in vivo, either by mutagenic primers or by erroneous DNA synthesis. Thus, choices largely depend on the experimental needs and resources of the investigator. PMID- 9417772 TI - Bay or baylike regions of polycyclic aromatic hydrocarbons were potent inhibitors of Gap junctional intercellular communication. AB - Many polycyclic aromatic hydrocarbons (PAHs) are known carcinogens, and a considerable amount of research has been devoted to predicting the tumor initiating potential of PAHs based on chemical structure. However, there has been little research into the effects of PAHs on the epigenetic events of tumor promotion and no structural correlation has been made thereof. Gap junctional intercellular communication (GJIC) activity was used in this study as an epigenetic biomarker to determine the structure-activity relationships of twelve different PAHs. The PAHs used were naphthalene, 1-methylnaphthalene, 2 methylnaphthalene, anthracene, 1-methylanthracene, 2-methylanthracene, 9 methylanthracene, 9, 10-dimethylanthracene, phenanthrene, fluorene, 1 methylfluorene, and fluoranthene. Results showed that PAHs containing bay or baylike regions inhibited GJIC more than did the linear PAHs. The nonnaphthalene PAHs were not cytotoxic as determined by a vital dye uptake assay, but the naphthalene compounds were cytotoxic at the higher doses, indicating that the down regulation of GJIC by these naphthalenes could be a consequence of general membrane damage. Inhibition of GJIC by all the inhibitory PAHs was reversed when the cells were refreshed with PAH-free growth medium. Inhibition of GJIC occurred within 0.5-5 min and correlated with the aqueous solubility of the PAHs. The present study revealed that there are structural determinants of epigenetic toxicity as determined by GJIC activity. PMID- 9417774 TI - A long-lived, highly luminescent Re(I) metal-ligand complex as a biomolecular probe. AB - A highly luminescent rhenium (I) metal-ligand complex [Re(bcp)(CO)3(4 COOHPy)](ClO4), where bcp is 2,9-dimethyl-4,7-diphenyl-1,10-phenanthroline and 4 COOHPy is isonicotinic acid, has been synthesized and characterized. High quantum yields (> 0.5) and long excited-state lifetimes (0.3-10 micronseconds) in fluid solutions at room temperature were found for this complex, with remarkable emission sensitivity to microenvironment. This compound also displays highly polarized emission with a maximum anisotropy near 0.3 in the absence of rotational diffusion. This Re complex was conjugated to several biomolecules, including the proteins human serum albumin and bovine immunoglobulin G, as well as an amine-containing lipid. When bound to a protein or lipid, the decay time is near 3 microseconds and the quantum yield is approximately 0.12 in aqueous oxygenated solution at room temperature. This compound's unique spectral properties along with its conjugatability allowed us to utilize it as biomolecular probe in a variety of environments. PMID- 9417775 TI - An expression cloning method to identify monomeric GTP-binding proteins by GTP overlay. AB - We have developed a method for identifying monomeric GTP-binding proteins that is based on probing plasmid expression libraries with [alpha-32P]GTP. The method involves the production of nitrocellulose replica filter lifts from a plasmid cDNA expression library and treatment of the filters with chloroform vapor to lyse the Escherichia coli and to denature and inactivate endogenous E. coli GTP binding proteins, thus allowing the direct identification of cDNA clones which encode Ras-like small GTP-binding proteins by ligand blotting. Using this procedure we have cloned a series of small Ras-like GTP-binding proteins from human retina. The method relies on a functional test, ligand specificity of the expressed proteins, to identify candidate molecules. This results in the isolation of predominantly full-length cDNA clones without relying on DNA sequence similarity. Thus, this method may be particularly useful for the cloning of novel Ras-related GTP-binding proteins which share limited sequence similarity with previously identified members of the Ras superfamily. PMID- 9417776 TI - Characterization of apical and basolateral plasma membrane domains derived from cultured rat cholangiocytes. AB - Cholangiocytes, the epithelial cells that line intrahepatic bile ducts, are composed of plasma membranes with discrete apical (lumenal) and basolateral domains that contain different channels, transporters, and receptors. In recent work, we developed a long-term, primary culture system of normal rat cholangiocytes (NRC). Our aims here were to prepare and characterize apical and basolateral plasma membrane vesicles from NRC. Using serial isopycnic centrifugation on sucrose gradients, we generated separate apical and basolateral plasma membrane vesicles. We characterized these vesicles by transmission electron microscopy, specific marker enzyme assays, and immunoblotting; we also determined the percentage of sealed vesicles and their intravesicular volume and sidedness using biochemical approaches. Our results showed that vesicles derived from the apical and basolateral plasma membrane domains of NRC were highly purified, predominately sealed, right-side-out vesicles that differed in size and lipid and protein composition. This experimental model represents a novel tool that will be useful for additional functional studies on the channels, transporters, and receptors differentially distributed in the plasma membrane of biliary epithelia. PMID- 9417777 TI - Determination of degradation rates of transfer and ribosomal ribonucleic acids in cultured rat hepatocytes by measuring N6-threoninocarbonyladenosine, dihydrouridine, and pseudouridine in medium using high-performance liquid chromatography. AB - Modified ribonucleic acid catabolites excreted into the medium by primary cultures of rat hepatocytes (2.3 +/- 0.42 x 10(6) cells/dish) during a 24-h cultivation period were quantified by reversed-phase high-performance liquid chromatography (fmol/10(3) cells): 613 +/- 81 dihydrouridine, 46 +/- 6 N6 threoninocarbonyladenosine, 1879 +/- 220 pseudouridine. On the basis of these excretion rates and the average frequency of occurrence of these modified ribonucleosides per cytoplasmic transfer ribonucleic acid (residues: 2.6 dihydrouridine, 0.22 N6-threoninocarbonyladenosine, 3 pseudouridine) as well as per cytoplasmic ribosomal ribonucleic acid (residues: 95 pseudouridine), the degradation rates of transfer and ribosomal ribonucleic acids were calculated. The degradation rate of transfer ribonucleic acid (fmol/10(3) cells/24 h) was 236 +/- 31 (via dihydrouridine) and 211 +/- 28 (via N6-threoninocarbonyladenosine) and that of ribosomal ribonucleic acid (fmol/10(3) cells/24 h) was 13.1 +/- 1.7 (via pseudouridine and N6-threoninocarbonyladenosine). PMID- 9417778 TI - An expression system for mammalian amino acid transporters using a stably maintained episomal vector. AB - Despite its versatility and effectiveness in numerous studies, the vaccinia/HeLa cell expression model may not be optimal for the study of all transport proteins. To evaluate an alternative expression model for amino acid transport Systems ASC and X-AG, the mRNA content and transport activity encoded by human hippocampal ASCT1 cDNA and rat hippocampal EAAC1 cDNA, respectively, were measured in pDR2 cDNA-transfected human embryonic kidney 293 cells made competent by stable transfection with the Epstein-Barr neutral antigen-1 (EBNA-1) cDNA (293c18 cells) to evaluate the EBNA-1/293c18 expression system. The results show that (i) the EBNA-1/293c18 expression system results in a larger increase over background of Systems ASCT1 (6.4x) and EAAC1 (39x) transport activity than does the vaccinia/HeLa expression system (2.6x and 22x, respectively); (ii) transfection and hygromycin B selection for the pDR2 vector do not affect the endogenous transport velocities of Systems ASC, X-AG, or A; and (iii) the endogenous transport velocities of Systems ASC and X-AG in 293c18 cells were not affected by the expression of exogenous EAAC1 or ASCT1. We conclude that the EBNA-1/293c18 cell expression model represents a useful transient expression regimen to characterize mammalian amino acid transport proteins, especially for transporters that may exhibit relatively low activity in transient expression systems lacking a selection mechanism. PMID- 9417779 TI - A method to study kinetics of transnitrosation with nitrosoglutathione: reactions with hemoglobin and other thiols. AB - The rate of protein S-nitrosylation, a reversible process by which S-nitroso thiol (RS-NO) compounds exchange the NO+ moiety with protein SH groups, is essentially governed by two factors, the pK alpha and the accessibility of the protein sulfhydryl. A useful method of following transnitrosation kinetics of various protein and nonprotein SH compounds with GS-NO is described. When the reaction is carried out in the presence of 1-chloro-2,4-dinitrobenzene and glutathione transferases, the rate of RS-NO formation (RSH + GS-NO-->RS-NO + GSH) can be monitored by spectrophotometry at 340 nm in terms of the enzymatic conversion of GSH to a GS conjugate. Unlike methods based on NO release from the S-NO bond, this procedure is rapid and accurate and requires relatively small amounts of thiols. The second order rate constants of S-nitrosylation of human and rat oxy- and deoxyhemoglobin of BSA and other thiols were calculated by this method which confirmed previous results reported in the literature. PMID- 9417780 TI - Isolation of gangliosides by cloud-point extraction with a nonionic detergent. AB - A rapid, simple, and efficient chloroform/methanol-free method of isolating gangliosides is described. The nonionic polyoxyethylene detergent, hexaethyleneglycol mono-n-tetradecyl ether (C14EO6), forms clear micellar solutions in water, but two-phase separation can be achieved by centrifugation in the presence of ammonium sulfate at room temperature. A mixture of pure gangliosides, metabolically labeled gangliosides obtained from cultured hippocampal neurons, and gangliosides from rat cerebellar tissue were quantitatively recovered in the detergent-rich upper (coacervate) phase, with a partition coefficient K > 60. Gangliosides were subsequently separated from the detergent using an Iatrobead column prior to analysis by thin-layer chromatography. The procedure described here is as efficient as other methods of ganglioside extraction, such as that using chloroform/methanol/water/pyridine, but is less time-consuming inasmuch as extraction, purification, and TLC analysis can be completed within 1 day. PMID- 9417781 TI - A continuous fluorometric assay for the feline immunodeficiency virus protease. AB - A novel fluorogenic substrate for continuous feline immunodeficiency virus (FIV) protease (PR) assay was developed in which 2-aminobenzoic acid (Abz) and p nitrophenylalanine (F(NO2)) were used as the fluorescent donor and acceptor, respectively. The 14-amino-acid fluorogenic substrate of sequence RALTK(Abz) VQ approximately F(NO2)VQSKGR (approximately indicates cleavage site) was modeled after a naturally occurring FIV PR capsid/nucleocapsid cleavage site in the gag polyprotein. The 2-aminobenzoyl group was attached to the epsilon amino group of a lysine (K(Abz)) in position P3 and the F(NO2) is in position P1' in order to promote efficient intramolecular quenching prior to cleavage by FIV PR. We measured a K(m) of 33 +/- 6 microM and a kcat of 0.29 +/- 0.02 s-1 for the enzymatic hydrolysis of this fluorogenic substrate by FIV PR under the conditions of our assay (0.05 M sodium citrate/0.1 M sodium phosphate buffer, pH 5.25, 0.2 M NaCl, 0.1 mM EDTA, and 1 mM dithiothreitol). This assay affords a rapid and convenient means for quantitating FIV PR activities and promises to be useful for judging the relative strength of inhibitors. PMID- 9417782 TI - Production of a soluble cyclin B/cdc2 substrate for cdc25 phosphatase. AB - Study of the function and regulation of the important cell cycle regulator cdc25 phosphatase has been hampered by the lack of a sensitive and specific substrate and assay. Here we report the production of a specific and sensitive substrate for the cdc25 phosphatase. The substrate is human cyclin B1/cdc2 phosphorylated on the inhibitory Thr14 and Tyr15 residues and activating Thr161 on cdc2, and is relatively simple to produce from readily available materials. The assay is based on the cdc25-specific dephosphorylation and activation of the phosphorylated cyclin B1/cdc2 substrate (PY15), using the increased histone H1 kinase activity of the activated PY15 as a read-out of cdc25 activity. PMID- 9417783 TI - Application of capillary electrophoresis to the measurement of oligonucleotide concentration and purity over a wide dynamic range. AB - Synthetic oligonucleotides rarely contain 100% of the full-length sequence due, in part, to the failure sequences produced during synthesis. In this paper, a method is described for the determination of both the concentration and the purity of oligonucleotides, utilizing capillary electrophoresis with a deoxyribo nucleoside triphosphate as an internal standard. This method is advantageous for several reasons: (a) the wide dynamic range allows for the analysis of samples without the need for dilutions; (b) a small sample size is used for analysis; (c) capillary electrophoresis is automatable which allows for high throughput; and (d) all of the samples are analyzed at the same run temperature which aids in reproducibility and consistency between runs performed at different times. PMID- 9417784 TI - Analysis of the Morgan-Elson chromogens by high-performance liquid chromatography. AB - The Morgan-Elson method for quantitative N-acetylhexosamine analysis is a two step procedure comprising alkali treatment of the sugar and subsequent condensation of the resulting chromogens with p-dimethylaminobenzaldehyde (Ehrlich's reagent) to yield a colored product. In the present investigation, the products formed in the first step of the procedure were analyzed by high performance liquid chromatography (HPLC) on a reversed-phase (C18) column, which was eluted with a water-methanol gradient; the absorbance of the effluent was monitored at 229 nm. The profile generated from alkali-treated N acetylglucosamine exhibited two major peaks, in a ratio of approximately 2.5:1, which accounted for 94% of the total peak area. A third peak, accounting for 3% of the peak area, was eluted in an intermediate position, and several smaller peaks were also observed. The three predominant components, isolated by preparative HPLC, all gave a purple color on addition of Ehrlich's reagent, indicating that they were Morgan-Elson chromogens. The HPLC profile of alkali treated N-acetylmannosamine was identical to that of the products generated from N-actylglucosamine, as was expected because of the elimination of the asymmetry at C-2 during formation of the chromogens. N-Acetylgalactosamine yielded two major peaks, which were eluted in the same positions as the two major products formed from N-acetylglucosamine, but the intermediate peak seen in the N acetylglucosamine pattern was absent. The HPLC procedure allowed detection of as little as approximately 25 ng of N-acetylglucosamine and may therefore be of value as an alternative to the complete Morgan-Elson procedure when only small amounts of sample are available for quantitative analysis. PMID- 9417785 TI - Determination of inorganic phosphate by coupling thymidine phosphorylase and reversed-phase high-performance liquid chromatography: application to tonoplast pyrophosphatase activity. AB - We developed an HPLC method for the measurement of inorganic phosphate using thymidine phosphorylase (EC 2.4.2.4). This enzyme catalyzes the phosphorolysis of thymidine to 2-deoxyribose 1-phosphate and thymine. Thymine release was measured at 265 nm after separation by reverse-phase HPLC. The assay was sensitive enough to detect as little as 10 pmol of Pi. The response to the phosphate concentration was linear from 1 to 100 microM. The value of this method was demonstrated in an analysis of the kinetics of Pi release from PPi in the presence of Catharanthus roseus tonoplast pyrophosphatase. PMID- 9417786 TI - Comparison of results of various methods used to determine the extent of modification of methoxy polyethylene glycol 5000-modified bovine cupri-zinc superoxide dismutase. AB - The protein bovine cupri-zinc superoxide dismutase (SOD) was modified by the reaction of lysine residues with an active ester of methoxy polyethylene glycol 5000 (PEG). The extent of modification was determined by capillary zone electrophoresis, matrix-assisted laser desorption/ionization mass spectrometry, Fourier-transform infrared spectroscopy, and Raman spectroscopy and after removing PEG by alkaline hydrolysis of the linkages to SOD followed by quantification of the released PEG using gel-permeation chromatography. There was generally good agreement among the results obtained by these techniques on a typical sample of the PEG-modified protein. The results, extent of protein modification, determined by the preceding methods were compared to that found from the classical trinitrobenzene sulfonic acid (TNBS) procedure. In addition to providing alternatives to the TNBS procedure, results from the described methodologies strongly suggest that the extent of modification determined from the TNBS procedure is overestimated. PMID- 9417787 TI - Magnetic bead enzyme-linked immunosorbent assay (ELISA) detects antigen-specific binding by phage-displayed scFv antibodies that are not detected with conventional ELISA. AB - An efficient means for the detection of antigen-specific binding by phage displayed antibodies would facilitate the selection of such phage, especially from libraries with large repertoires of V-genes. We report the development and characterization of a magnetic bead phage ELISA which detects antigen binding phage which could not be detected by conventional ELISA. We were attempting to select phage binding to the oncodevelopmental antigen, heat-stable alkaline phosphatase (HSAP). Although there was an obvious enrichment in the phage titers after successive rounds of selection, we were unable to detect antigen-binding phage by ELISA on a plastic surface. However, ELISA with a suspension of superparamagnetic particles covalently conjugated to HSAP effectively identified antigen-binding phage after the fourth round of selection. This method could also detect antigen-specific binding of individual phage clones. Some of the phage clones bound to either amino- or carboxy-terminal-conjugated HSAP, perhaps reflecting the differences in the exposed epitopes. It is suggested that a sensitive method such as magnetic bead phage ELISA be tried before declaring a phage selection as unsuccessful or concluding that a phage clone does not bind antigen. PMID- 9417788 TI - Determination of melanin in human hair by photoacoustic spectroscopy. AB - A highly sensitive photoacoustic spectroscopy for the determination of melanin in human hair has been developed. Both synthetic melanin and human hair were treated with 50% sulfuric acid. The melanin suspension was passed through the membrane filter to collect melanin, and the filter was transferred into a homemade photoacoustic cell, followed by measurement of the photoacoustic signals produced by 10 mW He-Ne laser beam. The good correlation between the method described in this paper and absorption spectrophotometry was confirmed quantitatively. The linearity observed between the photoacoustic signal and the weight of samples was in the range of 0.1-20 micrograms for synthetic melanin and 10-800 micrograms for human hair, with detection limits of 5 ng and 0.38 microgram, respectively. PMID- 9417789 TI - The simultaneous measurement of low rates of CO2 and O2 exchange in biological systems. AB - An instrument for measuring low rates of biological O2 exchange using an open flow gas analysis system is described. A novel differential O2 sensor that is capable of measuring as little as 0.4 Pa O2 against a back-ground of ambient air (20,900 Pa O2), yet has a dynamic range of +/- 2000 Pa O2 (i.e., +/- ca. 2% O2) is described. Baseline drift was typically less than 0.025 Pa min-1. The differential O2 sensor was incorporated into a respiratory quotient/photosynthetic quotient analyzer that contained other environmental sensors for atmospheric pressure, absolute O2 and CO2 concentration, temperature of the differential O2 sensor block, and differential pressure between reference and sample streams. Protocols for how these sensors can be used to calibrate the differential O2 sensor and to improve its stability with time are described. Together, the differential O2 sensor, the environmental sensors, and the simple calibration techniques allow for simultaneous, noninvasive, and accurate measurements of O2 and CO2 exchange in tissues with metabolic rates as low as about 0.1 mumol O2 or CO2 h-1. Example data are provided in which O2 differentials of 3 to 41 Pa O2 were measured in an open-flow system. PMID- 9417790 TI - Highly sensitive time-resolved fluoroimmunoassay of human immunoglobulin E by using a new europium fluorescent chelate as a label. AB - A new europium fluorescent chelate, 4,4'-bis(1",1",1",2",2",3",3"-heptafluoro 4",6"-hexanedione-6"-yl) - chlorosulfo-o-terphenyl (BHHCT)-Eu3+, was used as a label for highly sensitive time-resolved fluoroimmunoassay of human IgE. Two assay formats were employed in the analysis. In the first format, an immunoconjugate of rabbit anti-human IgE antibody-human IgE-biotinylated goat anti-human IgE antibody-BHHCT-Eu(3+)-labeled SA (or BHHCT-Eu(3+)-labeled BSA-SA; BSA, bovine serum albumin; SA, streptavidin) was used for measurement. The method gives the detection limits of 3.6 x 10(-2) IU/ml (labeled SA) and 1.1 x 10(-2) IU/ml (labeled SA-BSA). In the second format, an immunoconjugate of goat anti human IgE antibody-human IgE-rabbit anti-human IgE antibody-biotinylated goat anti-rabbit IgG antibody-BHHCT-Eu(3+)-labeled SA (or BHHCT-Eu(3+)-labeled BSA-SA) was used for measurement. The detection limits of these methods are 3.0 x 10(-3) IU/ml (labeled SA) and 1.5 x 10(-3) IU/ml (labeled BSA-SA). The above detection limits are one to two orders of magnitude lower than those of the conventional radioimmunoassay and enzyme immunoassay. The CV of the present method is less than 7%, and the recovery is in the range of 85-105% for serum samples. PMID- 9417791 TI - High-level overexpression of yeast elongation factor 3 and detailed kinetic analysis using a coupled spectrophotometric assay. PMID- 9417792 TI - An automated liquid-phase assay for quantitation of protein tyrosine kinase activity. PMID- 9417794 TI - Reflections on risk and resilience in adolescence. PMID- 9417793 TI - Separation of polyphosphoinositides using normal-phase high-performance liquid chromatography and evaporative light scattering detection or electrospray mass spectrometry. PMID- 9417795 TI - Parent-child relationships as a protective factor in preventing adolescents' psychosocial risk in inter-racial adoptive and non-adoptive families. AB - This study aims to compare family relations in inter-racial adoptive and non adoptive families with a late adolescent and to examine the extent to which family relations constitute a protective factor in preventing adolescents' psychosocial risk. The sample consisted of 103 inter-racial adoptive families and 150 non-adoptive families with a child aged between 16 and 19 years. Subjects were requested to complete a self-report questionnaire. The results suggest the existence of a different relational configuration in inter-racial adoptive and non-adoptive families. They also show that the father-child relationship and the mother-child relationship play different roles in preventing the adolescent's maladjustment in inter-racial adoptive and non-adoptive families. PMID- 9417796 TI - Adolescent friendships mediating childhood adversity and adult outcome. AB - This interview-based study compares the friendships of 50 girls, aged 15-16, identified on the basis of their childhood experiences as being at-risk for difficulties in early adult partnerships, with the friendships of 50 girls of the same age from an inner-city school. Key differences in the features of both romantic and non-romantic adolescent friendships between the two groups of girls give a clearer understanding of the processes linking childhood adversity and poor adult outcome. PMID- 9417797 TI - Behaviour problems and relationships with family and peers during adolescence. AB - This study examines the relationship between (a) characteristics of adolescents' social networks, personal resources, and environmental risks; and (b) adolescents' behaviour problems. A socio-ecological perspective was used to determine whether social support and perceived conflict operate as risk or protective factors. Three samples of adolescents were studied: 63 adolescents in residential care; 29 in day treatment; and 63 who had had no contact with professional care. Interrelations were explored by means of structural equation modelling. Social support in family and peer group may operate as a risk factor or a protective factor depending on other risk factors in these subsystems. Perceived conflict is related to behaviour problems, although the influence differs depending on environmental risk factors and the type of behaviour problems. PMID- 9417798 TI - Protective and risk effects of peer relations and social support on antisocial behaviour in adolescents from multi-problem milieus. AB - This article addresses the relation between antisocial behaviour and social resources in a 2-year longitudinal study of 100 high-risk adolescents in residential care. Problem behaviour was measured with the Externalizing Scale of the Youth Self Report. Social resources were recorded using semi-structured methods. Hierarchical regression analyses showed interactions suggesting that the same variables can fulfil risk as well as protective functions: clique membership and satisfaction with social support fostered behavioural continuity. In contrast, a lack of social embeddedness had a risk effect for well-adapted adolescents and a protective effect for the deviant ones. Social resources were more influential in girls. Theoretical implications and methodological problems are discussed. PMID- 9417799 TI - Resilience, desistance and delinquent career of adolescent offenders. AB - This study examines resilience and desistance from delinquent behaviours and attempts to identify factors which predict persistent or increased or decreased delinquency between adolescence and early childhood. A sample of 363 young people was obtained from the population of five public institutions in 1987 and 1992. Their delinquent trajectories were described on the basis of legal records. These suggested that resilience is a rare phenomenon and is associated with stable relationships, absence of diagnostic label and good adaptation to the institution. Two axes were identified which can be used to describe the population, the family background and the psychological characteristics of the individual. Information which predicted desistance from a delinquent career was identified by means of a stepwise multiple regression. Analyses were conducted separately for each age group to take account of differences in time after the placement. The results indicated that there are important age-related differences in the characteristics which influence desistance or risk. They also show length of stay in an institution to be a predictor. Desistance from further delinquency seems to depend on the time spent in the residential environment associated with an increase of guilt, an improvement of self-image and of attachment to one or more other people. PMID- 9417800 TI - The self-concept of societally vulnerable and delinquent boys within the context of school and leisure activities. AB - The central question in this article is: "Have delinquent and non-delinquent boys different self-concepts?" The idea is that it is necessary to distinguish between different domains that are relevant to adolescent boys, that is school and leisure time, and between conventional and non-conventional characteristics. An instrument was constructed to measure the school/conventional orientated and leisure/non-conventional orientated self-concept. The research group consisted of 219 adolescent boys. Girls were not included. The results show that most delinquent boys have a leisure/non-conventional orientated self-concept, whilst most non-delinquents, as well as most boys who are not involved in the peer group, have a school/conventional orientated self-concept. The implications of the research and suggestions for other research are discussed. PMID- 9417801 TI - Adolescents' judgements of the seriousness of disruptive behaviour at school and of the sanction appropriate for dealing with it. AB - Disruptive behaviour is a daily event in many secondary schools. The goal of this study is to discriminate adolescents' attitudes towards disruptive behaviour in school from their attitudes towards the authors of such behaviour. The study was conducted using a questionnaire referring to situations which disrupt the class. Adolescents (n=172) estimated the seriousness of the action involved in each situation and said how those responsible should be dealt with. The factors taken into account were: the subjects' gender, whether s/he is actively involved in disruptive behaviour or not, the gender of the central character and the nature of the disturbance. The results show that: (1) judgement of the seriousness of the behaviour is more severe than the sanctions thought to be appropriate. This is particularly so for violent disruptions; (2) pupils who are themselves disruptive are more tolerant than non-disruptive pupils, both in their judgement of the seriousness of action and in their ideas about sanctions. PMID- 9417802 TI - Abnormal expansion of peripheral gamma delta T cells in patients with neurologic disorders. AB - The proportions and the variable region usages of peripheral gamma delta T cells were investigated in 205 patients with various neurologic disorders. Flow cytometric analysis was performed with monoclonal antibodies against C delta, V delta 1, V delta 2. V delta 1-J delta 1/J delta 2, and V gamma II (9) epitopes. Further analysis was carried out with reverse transcription-polymerase chain reaction (RT-PCR) using variable region-specific primers. The proportion of gamma delta T cells in 33 patients with cerebral palsy (CP) (mean +/- SD; 11.1 +/- 11.0%) was significantly higher than that in 35 normal controls (5.6 +/- 2.6%) (p < .01). Of 205 patients, persistent gamma delta T cell expansion over 15% was observed in 15 patients (M/F = 13/2). Nine of these 15 were patients with CP. Six of the 9 with CP had neither perinatal nor postnatal events related to the neurologic abnormalities. None of the 9 with CP had active infections or autoimmune disorders that could induce gamma delta T cell expansion. Of the 15, 13 had V gamma II/V delta 2-dominant expansion and 2 showed V delta 1-dominant expansion. Unusual expansions of V delta 4 and V delta 6, which were not usually found in peripheral blood, were detected by RT-PCR in one case with CP and V delta 1-dominant expansion. This study showed subclinical immunologic abnormalities with marked and persistent gamma delta T cell expansion in patients with neurologic disorders, especially CP. These results might indicate the existence of neuroimmunologic disorder(s). PMID- 9417803 TI - Acetylcholinesterase monoclonal antibody-induced sympathectomy: effects on immune status and acute morphine-induced immunomodulation. AB - The present study examined the role of the sympathetic nervous system (SNS) in immunomodulation by using the acetylcholinesterase monoclonal antibody (AChE mAb) induced sympathectomy model. As part of this investigation, the effects of AChE mAb treatment on the immune alterations produced by acute morphine treatment also were explored. Experimental rats received tail vein injections of murine monoclonal IgG2b antibodies against rat brain acetylcholinesterase, which produce a destruction of cholinergic, sympathetic preganglionic neurons and a resultant decrease in sympathetic activity. Control rats received tail vein injections of murine IgG antibodies, which do not affect sympathetic preganglionic neurons or sympathetic activity. One week after antibody treatment, rats received a subcutaneous injection of 15 mg/kg morphine or the saline vehicle. One hour after the morphine or saline injections, rats were sacrificed and immune assays were conducted. AChE mAb treatment increased the mitogen-stimulated proliferation of splenic T cells and interleukin-2 (IL-2) production by stimulated splenocytes, indicating that these immune measures are sensitive to the AChE mAb-induced alteration in sympathetic function. Treatment with AChE mAb did not alter the mitogen-stimulated proliferation of splenic B cells or blood T cells, splenic natural killer (NK) cell activity, or the production of interferon-gamma (IFN gamma) by stimulated splenocytes, indicating that these immune measures are relatively insensitive to the AChE mAb-induced alteration in sympathetic activity. The AChE mAb-induced alteration in sympathetic activity did not affect the suppressive effects of acute morphine treatment on the mitogen-stimulated proliferative response of splenic T and B cells and blood T cells, splenic NK cell activity, and the production of IFN-gamma and IL-2 by stimulated splenocytes. PMID- 9417804 TI - Involvement of interleukin-1 beta, nerve growth factor, and prostaglandin-E2 in the hyperalgesia induced by intraplantar injections of low doses of thymulin. AB - The effect of various doses of intraplantar thymulin injection, on nociceptive thresholds, in the hind paw of rats was assessed using different pain tests. As little as 0.5 ng thymulin resulted in localized mechanical hyperalgesia as assessed by the paw pressure test and thermal hyperalgesia as assessed by the paw immersion, hot plate, and tail flick tests. The highest dose of thymulin (10 ng) reduced both paw pressure and paw immersion latencies in the noninjected paw also. Thymulin (5 ng) also resulted in significant elevation in the levels of interleukin-1 beta (IL-1 beta) and nerve growth factor (NGF) levels in the injected paw. Both dexamethasone and indomethacin reversed thymulin-induced hyperalgesia. Also interleukin-1 receptor antagonist (IL-1ra) and a polyclonal anti-NGF antiserum significantly reduced thymulin-induced hyperalgesia. On the other hand, the tripeptide lys-D-pro-val (known to antagonize IL-1 beta and PGE2 induced hyperalgesia) reversed the hyperalgesia due to thymulin. In conclusion, thymulin induces localized hyperalgesia which is mediated by PGE2-dependent mechanisms and this pathway could be either partially dependent on or totally independent of IL-1 beta mechanisms. PMID- 9417805 TI - Caregiving to very low birthweight infants: a model of stress and immune response. AB - Mothers of preterm, very low birthweight (< or = 1500 g; VLBW) infants experience the stress of caring for small, fragile infants at the same time that they are recovering from the relative immunosuppression of pregnancy and when many health behaviour changes (e.g., nutrition) occur which also may influence immune status. The purpose of this study was to examine changes in anxiety and depression and in health behaviors, as well as lymphocyte proliferation and natural killer cell activity in mothers of preterm, VLBW infants compared to mothers of healthy term infants. Mothers of preterm VLBW infants have decreased in vitro lymphocyte response to mitogens compared to mothers of healthy term infants over time, and this difference could not be explained by anxiety, depression, or health behaviors. However, among mothers of VLBW infants, anxiety was related to decreased lymphocyte proliferation response at 1 month postpartum. There was no relationship between maternal depression and lymphocyte proliferative response in mothers of term infants. Natural killer cell activity did not differ between the two groups of mothers, nor was there a relationship between natural killer cell activity and maternal anxiety, depression, or health behaviors. Thus, lymphocyte proliferative response to mitogens may be an important biologic market of increased stress in mothers of VLBW infants in the first couple of months postpartum. PMID- 9417806 TI - The effect of exercise on lymphocyte redistribution and leucocyte function in asymptomatic HIV-infected subjects. AB - This study was undertaken to examine the responsiveness of circulating leucocyte and lymphocyte populations to the physiological demands of exercise in asymptomatic HIV-infected subjects with CD4+ counts greater than 500/microliter. Thirteen subjects infected with HIV and 14 control subjects underwent 20 min of defined moderate exercise at estimated 65% of their maximal ventilatory capacity on a bicycle ergometer. Blood samples were obtained for serum cortisol, norepinephrine, lymphocyte subsets (CD4, CD8, CD19, CD16-CD56), and phagocytic function at rest immediately after exercise and 20 min following the cessation of the exercise. The HIV-infected subjects had increased circulating concentrations of CD8 cells (p = .007) and CD16-CD56+ NK cells (p = .02) in response to the exercise, whereas the control group did not. There was a greater increase in monocyte respiratory burst activity following recovery from exercise in the control subjects (p = .016) but not in the HIV-infected subjects. The control subjects experienced an increase in serum cortisol in response to the exercise (p = .006), but the HIV-infected subjects did not. Our results show that the changes in the distribution and function of circulating leucocytes and adrenal neuroendocrine responses to moderate exercise differ in asymptomatic HIV-infected and control subjects. PMID- 9417807 TI - Differential effect of lipopolysaccharide on food hoarding behavior and food consumption in rats. AB - Experimental studies assessing the suppressing effect of lipopolysaccharide (LPS) on feeding behavior have focused exclusively on the ingestive component of this behaviour without taking into account its appetitive component. The appetitive sequence of feeding behavior regroups activities animals engage in to gain access to food without necessarily eating it. The objective of the present study was to compare the effects of LPS on food intake and food hoarding. Rats were given the possibility to access food during a 30-min daily session in an apparatus consisting of a cage connected to an alley with free food at its end. Subjects were tested under different motivational levels for food hoarding: a first group (FS) received a food supplement to maintain stable body weight while a second group (noFS) did not receive such a supplement. LPS (250 micrograms/kg i.p.) dramatically decreased total food intake in rats from both groups whereas food hoarding was much less affected in LPS-treated rats from the noFS group. This expression of a still salient secondary motivation in LPS-treated rats which did not receive any food supplement can be interpreted to suggest the expression of an anticipatory feeding behavior along with a reduced immediate appetite. In addition, LPS had no effect, in rats from the noFS group, on the amount of food eaten after transport to the refuge. LPS-treated animals still appear to be able to adjust their defensive behavioral strategies with regard to their needs and capacities. These findings support the adaptive value of the behavioral changes displayed by LPS-treated animals. PMID- 9417808 TI - Identification of a novel glucocorticoid response unit (GRU) in the 5'-flanking region of the mouse IL-2 receptor alpha gene. AB - Glucocorticoid hormones inhibit the production of IL-2 and upregulate mouse interleukin 2 receptor alpha (IL-2Ralpha) gene expression in T cell lines by increasing its transcription rate. Now, the authors have used functional approaches to search for regulatory elements present in the 5'-flanking region of the IL-2Ralpha gene responsible for this effect. An important regulatory region was detected between -1382 and -1100 bp from the transcription initiation site. Within this region the authors characterized two 20 bp long cis-acting regulatory elements, named G1 and G2, which are involved in the modulation of the expression of the IL-2Ralpha gene by glucocorticoids. G1 contains a relatively well conserved GRE half palindrome site, able to bind a partially purified glucocorticoid receptor but giving rise to an unstable complex. The G2 regulatory element contains no consensus sequences of binding sites for GR nor for any other described transcriptional factors but is able to form complexes with factors present in liver or T cells. Whereas G1 or G2 alone were unable to induce a glucocorticoid-response, the contiguous presence of G1 and G2 gave rise to an efficient response. Therefore, it is postulated that the glucocorticoid-induction of IL-2Ralpha gene is mediated, at least partly, through G1 and G2 elements which constitute a novel multicomponent glucocorticoid response unit (GRU). PMID- 9417810 TI - Expression of monocyte chemotactic protein-3 in human monocytes exposed to the mycobacterial cell wall component lipoarabinomannan. AB - Monocyte chemotactic protein-3 (MCP-3) is a C-C chemokine which interacts with the CCR1, CCR2 (MCP-1) and CCR3 receptors and has a distinct spectrum of action. The present study was designed to assess whether mycobacterial components were able to induce expression and production of MCP-3 in human monocytes. Mycobacterial lipoarabinomannan (LAM) induced expression of MCP-3 mRNA in human peripheral blood mononuclear cells. The non-mannose-capped version of lipoarabinomannan (AraLAM) was considerably more potent than the mannose-capped version ManLAM or the simpler version phosphatidylinositol mannoside (PLM). Among mononuclear cells, monocytes were responsible for LAM-induced MCP-3 mRNA expression. Whole mycobacteria (Mycobacterium bovis BCG) strongly induced MCP-3 expression. Pretreatment with actinomycin D abolished LAM-induced MCP-3 expression, whereas cycloheximide only partially reduced the expression. LAM induced MCP-3 expression was associated with the production of immunoreactive PTX3. Interleukin 10 (IL-10) and IL-13 inhibited the induction of MCP-3 by LAM. Thus mycobacterial cell wall components induced expression of MCP-3 in human monocytes. MCP-3, a chemokine active on mononuclear phagocytes, NK cells, T cells and dendritic cells, may be relevant to the induction and expression of immunity against mycobacteria. PMID- 9417809 TI - Negative regulation of the interleukin (IL)-2 receptor alpha chain (CD25) promoter region in human B lymphocytes. AB - Studies of the CD25 gene promoter region in T cells have revealed the presence of a 31-bp region, including an 11-bp negative regulatory element (NRE), which profoundly suppresses CD25 expression. This report illustrates that the same region acts as a negative regulator of CD25 expression in human B lymphocytes. Human B cells contain DNA-binding protein activities which bind specifically to an oligonucleotide equivalent to the 11-bp core region of the NRE, and stimulation of tonsillar B cells or Daudi Burkitt's lymphoma B cells with interleukin 4 (IL-4) results in loss of binding activity for oligonucleotides containing the NRE; in contrast, IL-4 enhanced binding activity for the NRE in the Jurkat T cell line. Transient transfection analyses using deletion mutants lacking both the 11-bp core NRE and both the NRE and an adjacent putative retinoic acid response element (RARE) motif illustrated that the NRE element is a functional suppressor of CD25 transcription in B cells. Thus, deletion of the NRE element increased the basal level of CD25 promoter activity and also conferred IL 4 inducibility on reporter gene expression in transiently transfected tonsillar B lymphocytes. PMID- 9417811 TI - The neuropeptide calcitonin gene-related peptide inhibits TNF-alpha but poorly induces IL-6 production by fetal rat osteoblasts. AB - Tumour necrosis factor alpha (TNF-alpha) and interleukin 6 (IL-6) are potent inflammatory cytokines produced by osteoblasts and whose contribution to bone loss occurring in oestrogen deficiency is well documented. Calcitonin gene related peptide (CGRP) is a neuropeptide abundantly concentrated in sensory nerve endings innervating bone metaphyses and periosteum suggesting that it controls bone homeostasis locally. Since CGRP was shown to inhibit TNF-alpha production by T cells and stimulate IL-6 expression by fibroblasts, this study was designed to investigate whether CGRP regulated TNF-alpha and IL-6 production by osteoblasts. We show that CGRP inhibits the production of TNF-alpha by both lipopolysaccharide (LPS)- and IL-1-stimulated fetal rat osteoblasts. Like CGRP, the cAMP agonists prostaglandin E2 (PGE2), dibutyryl cAMP (Bt2cAMP) and forskolin inhibit TNF-alpha production by osteoblasts. Exposure of osteoblasts to a high dose of phorbol myristoyl acetate (PMA) to deplete PKC activity abolished CGRP-mediated TNF-alpha suppression. In contrast with its potent inhibition of TNF-alpha production, we show that CGRP is a weak inducer of IL-6 when compared to PGE2, Bt2cAMP and forskolin. However, in presence of isobutylmethylxanthine (IBMX) CGRP stimulates the production of IL-6. Collectively, these data suggest that the inhibition of TNF-alpha CGRP is cAMP dependent and PMA sensitive and that the concentration of intracellular cAMP may be a regulatory mechanism for IL-6 expression in osteoblasts. PMID- 9417812 TI - Effects of unsaturated fatty acids on interleukin-1beta production by human monocytes. AB - Dihomogammalinolenic acid is derived from gammalinolenic acid, administration of which suppresses joint inflammation. It is reported here that interleukin 1beta (IL-1beta) production by human monocytes is enhanced markedly when cells are incubated 18-24 h with the polyunsaturated fatty acids dihomogammalinolenic acid (DGLA) and arachidonic acid (AA), then stimulated with lipopolysaccharide. Effects of eicosapentaenoic acid (EPA) on IL-1beta production are minimal, and palmitic acid (PA) does not influence IL-1beta production. PMID- 9417813 TI - Characterization of human recombinant interleukin 2 binding to heparin and heparan sulfate using an ELISA approach. AB - We have developed an enzyme-linked immunosorbent assay (ELISA) approach for the study of interactions between cytokines and glycosaminoglycans. This involves, as solid phase, a synthetic heparin-bovine serum albumin (BSA) complex in which the heparin is coupled via its reducing terminus to the protein using sodium cyanoborohydride. We have investigated the sensitivity and specificity of this experimental technique, employing antithrombin (AT III) and fibroblast growth factor 2 (FGF-2) as well-characterized heparin binding proteins. Using this ELISA method, we have established that human recombinant interleukin (IL-2) binds to heparin in a concentration-dependent manner. Soluble heparin competes for the binding of IL-2 to the complex with 50% inhibition at 5 microg/ml. This IC50 value provides an estimate of the binding constant of around 0.5 microM. This value is at least two orders of magnitude larger than that for the binding of IL 2 to its dimeric and trimeric cell surface receptors, but similar to that for binding to the IL-2 receptor beta polypeptide acting alone. Our ELISA shows that in addition to soluble heparin, fuciodan also competes for IL-2 binding, but chondroitin sulfate and dermatan sulfate are inactive. Of six heparan sulfates tested, only one highly sulfated preparation competed for IL-2. The interaction between IL-2 and heparin-like glycosaminoglycans is likely to be an important mechanism for retaining IL-2 close to its sites of secretion, thus giving rise to localized concentration gradients in the tissues. PMID- 9417814 TI - Specific changes in the pancreatic expression of the interleukin 1 family of genes during experimental acute pancreatitis. AB - Interleukin 1beta (IL-1beta) is produced in large amounts during acute pancreatitis and is believed to play a primary role in determining pancreatitis severity and the degree of pancreatic tissue destruction. This study was undertaken to characterize intrapancreatic production of IL-1beta and the remainder of the IL-1 family of genes during sterile acute pancreatitis. Moderate or severe necrotizing pancreatitis was induced by the intraperitoneal injection of a cholecystokinin analogue or the feeding of a choline deficient diet, respectively. Animals were killed during the progression of pancreatitis with severity scored by histological grading and serum amylase concentration. The expression of IL-1beta, IL-1 Receptor 1 (IL-1R1), Il-1R2, IL-1R antagonist (IL 1Ra), and ICE mRNA within the pancreas was examined by quantitative differential RT-PCR. Corresponding intrapancreatic and serum proteins were measured by enzyme linked immunosorbent assay (ELISA). There was constitutive expression of pancreatic IL-1R1, IL-1R2, IL-1Ra, and ICE but not IL-1beta. As pancreatitis developed, mRNA for IL-1beta, IL-1Ra, and ICE increased in parallel with the degree of pancreatitis severity (all P<0.001 vs baseline) while mRNA for both receptors remained stable (P=NS). Intrapancreatic and systemic IL-1beta and IL 1Ra protein also increased as pancreatitis developed (both P<0.001) with tissue levels being continuously greater than serum. This study demonstrated that sterile, endotoxin-free acute pancreatitis induces the upregulation of specific members of the IL-1 family of genes including production of large amounts of IL 1beta and its receptor antagonist within the pancreatic parenchyma. These changes are indicative of pancreatitis severity and are not model dependent. PMID- 9417815 TI - Nociceptive responses in interleukin-6-deficient mice to peripheral inflammation and peripheral nerve section. AB - The cutaneous nociceptive response threshold to mechanical and thermal stimulation, the development of hyperalgesia and plasma extravasation after subcutaneous injection of carrageenan and the development of autotomy behaviour after nerve section were assessed in interleukin-6-deficient (IL-6-/-) and age matched wild-type (IL-6+/+) mice. IL-6-/- mice had significantly lower response threshold to both mechanical and thermal stimulation in comparison to IL-6+/+ controls. Both IL-6-/- and IL-6+/+ mice developed hyperalgesia to mechanical and thermal stimulation after localized carrageenan injection, but the magnitude of the hyperalgesia was less in the IL-6-/- than in the IL-6+/+ controls. IL-6-/- mice also exhibited less plasma extravasation after carrageenan injection. No difference was noted between males and females in basal nociception and inflammatory hyperalgesia. However, female IL-6-/- mice exhibited autotomy behaviour, a sign of neuropathic pain, significantly more frequently and after a shorter interval following peripheral nerve injury than male IL-6-/- or male and female IL-6+/+ mice. It is suggested that IL-6-/- mice exhibited numerous changes in nociceptive responses compared to controls, some of which are sex related. The mechanisms of these changes in relation to null-mutation of the IL-6 gene and the influence of genetic background are discussed. 1997 Academic Press Limited. PMID- 9417816 TI - Systemic release of soluble TNF receptors after high-dose TNF in isolated limb perfusion. AB - Isolated limb perfusion (ILP) with high dose tumour necrosis factor (TNF), interferon gamma and melphalan (TIM) is an efficient treatment for patients with regionally advanced melanoma and sarcoma. In 44 patients, we determined the kinetics of soluble TNF receptors (sTNF-RI and RII) plasma concentrations, and correlated them with systemic TNF and interleukin 6 (IL-6) levels and shock. Seven patients treated conventionally by ILP without cytokine served as controls. Elevated levels of both sTNF-Rs were observed within 30 min after beginning of the TIM-ILP. A first peak of sTNF-Rs levels was observed 3 h after ILP and was followed by a rapid decrease reaching a nadir at 12-14 h post ILP. This first peak was followed by a second, long-lasting elevation of both sTNF-Rs levels persisting for 4 to 5 days after TIM-ILP. Patients treated by ILP without TNF/interferon gamma (IFN-gamma) had no detectable increase in either sTNF-Rs or in circulating TNF, demonstrating that the release of TNF-Rs was dependent upon the administration of TNF/IFN-gamma. High plasma levels of TNF and IL-6 were observed in patients that had more than 5% leakage during the TIM-ILP, but no significant correlation between TNF levels and the peak values of both sTNF-Rs was observed. The levels of TNF and IL-6 were, however, significantly related to each other. TNF systemic levels, but not sTNF-Rs concentrations, correlated significantly with the severity of the shock observed after TIM-ILP. Patients in which sTNF-RII concentration was in excess over circulating TNF, had no shock or grade I shock only, suggesting that sTNF-RII may play a protective, although limited, role in inhibiting activity of circulating TNF. PMID- 9417817 TI - Richard Paul Bunge, M.D. PMID- 9417818 TI - Richard Bunge. PMID- 9417819 TI - Central cord syndrome of cervical spinal cord injury: widespread changes in muscle recruitment studied by voluntary contractions and transcranial magnetic stimulation. AB - Muscle recruitment after central cord syndrome (CCS), a cervical spinal cord injury leading to a weaker motor function in the upper limbs versus the lower limbs, was examined in 14 individuals by means of voluntary muscle contractions and transcranial magnetic stimulation (TMS). Previously obtained data from able bodied (AB) and non-CCS spinal cord injured subjects were used for comparison. Surface EMG was recorded from as many as six pairs of affected muscles. Individual muscle EMG activity was scored from 0 to 5. Cortical stimulation was applied while subjects maintained a weak contraction in each muscle. When CCS subjects attempted to produce a maximal voluntary contraction of an isolated muscle, this frequently resulted in cocontraction of nonsynergists in the same limb or/and in other limbs. Although the EMG scores in both upper and lower extremity muscles improved within postinjury time, in general, the lower extremity muscles, particularly the distal ones, demonstrated better recovery than the upper extremity muscles. CCS and AB subjects showed a similar high probability of "well-defined" responses to TMS (amplitude >150 microV) in all studied muscles. In contrast, latencies to TMS-evoked motor responses were prolonged by significant amounts after CCS. The delays in muscle responses were not significantly different from those observed in subjects with more severe cervical injury. Despite improvement in EMG scores, repeated measurements of TMS evoked muscle response latencies in the same CCS subjects did not reveal significant shortening in central conduction latency. This argues against remyelination as an important contributor to the recovery process. PMID- 9417820 TI - Analysis of NMDA receptors in the human spinal cord. AB - NMDA receptors in postmortem human spinal cord were analyzed using [3H]MK-801 ligand binding and immunoblotting with NMDA receptor subunit-specific antibodies. The average KD for [3H]MK-801 binding was 1.77 nM with a Bmax of 0.103 pmol/mg. The EC50 for stimulation of -3H-MK-801 binding with L-glutamate was 0.34 microM. None of these parameters were affected by postmortem intervals up to 72 h. Immunoblotting of native NMDA receptors showed that NR1, NR2A, NR2C, and NR2D subunits could all be found in the human spinal cord of which NR1 was preferentially located to the dorsal half. Immunoprecipitation of solubilized receptors revealed that NR1, NR2C, and NR2D subunits coprecipitated with the NR2A subunit, indicating that native human spinal cord NMDA receptors are heteroligimeric receptors assembled by at least three different receptor subunits. These results provide a basis for the development of drugs selectively aimed at spinal cord NMDA receptors for the future treatment of spinal cord disorders. PMID- 9417821 TI - Muscle weakness, paralysis, and atrophy after human cervical spinal cord injury. AB - Muscle weakness and failure of central motor drive were assessed in triceps brachii muscles of individuals with chronic cervical spinal cord injury (SCI) and able-bodied controls. Electrical stimuli were applied to the radial nerve during rest and during triceps submaximal and maximal voluntary contractions (MVCs). The mean forces and integrated EMGs generated by SCI subjects during MVCs were significantly less than those produced by controls (P < 0.01), with 74 and 71% of muscles generating <10% control force and EMG, respectively. There was an inverse linear relationship between the evoked and voluntary forces (n = 32 muscles of SCI subjects) which, when extrapolated to zero evoked force, also showed significant whole muscle weakness for SCI compared to control subjects (P < 0. 01). Severe muscle atrophy was revealed which might reflect disuse and/or muscle denervation subsequent to motoneuron loss. Many triceps muscles of SCI subjects showed no force occlusion (n = 41) or were impossible to stimulate selectively (n = 61). Force was always evoked when the radial nerve was stimulated during MVCs of SCI subjects. The force elicited by single magnetic shocks applied to the motor cortex at Cz' during voluntary contractions of SCI subjects was also inversely related to the voluntary triceps force exerted (n = 18), but usually no force could be elicited during MVCs. Thus central motor drive was probably maximal to these muscles, and the force evoked during MVCs by below-lesion stimulation must come from activation of paralyzed muscle. SCI subjects also had significantly longer mean central nervous system (CNS) conduction times to triceps (P < 0.01) suggesting that the measured deficits reflect CNS rather than peripheral nervous system factors. Thus, the weak voluntary strength of these partially paralyzed muscles is not due to submaximal excitation of higher CNS centers, but results mainly from reduction of this input to triceps motoneurons. PMID- 9417822 TI - The astroglial response to Wallerian degeneration after spinal cord injury in humans. AB - We describe the changes exhibited by astrocytes in areas of Wallerian degeneration after spinal cord injury in humans using glial fibrillary acidic protein immunohistochemistry correlated to standard histology at time points ranging from 8 days to 23 years after injury. Astrocytes were slow to react; a slight increase in immunoreactivity was observed at 4 months. Over time they began to lose immunoreactivity in both the somata and the processes as the debris from the degenerative process was cleared. By 1 year after injury the staining intensity had decreased to levels which were lower than in normal areas of the cord. This hypointense staining persisted for at least 23 years after injury. These findings are significantly different from those observed in animal studies and emphasize the need for additional pathological studies of human spinal cord injury. PMID- 9417823 TI - Activated macrophage/microglial cells can promote the regeneration of sensory axons into the injured spinal cord. AB - A prominent role for phagocytic cells in the regenerative response to CNS or PNS injury has been suggested by numerous studies. In the present work we tested whether increasing the presence of phagocytic cells at a spinal cord injury site could enhance the regeneration of sensory axons from cut dorsal roots. Nitrocellulose membranes treated with TGF-beta or coated with microglial cells were cotransplanted with fetal spinal cord tissue into an injured adult rat spinal cord. Cut dorsal roots were apposed to both sides of the nitrocellulose. Four weeks later, animals were sacrificed and spinal cord tissue sections were processed for immunocytochemical detection of calcitonin gene-related peptide (CGRP-ir) to identify regenerated sensory axons. Adjacent sections were processed with the antibody ED-1 or the lectin GSA-B4 for detection of macrophage/microglial cells in association with the regrowing axons. Qualitative and quantitative data indicate a correlation between the pattern and extent of axonal regeneration and the presence of phagocytic cells along the nitrocellulose implant. Axonal regeneration could be experimentally limited by implanting a nitrocellulose strip treated with macrophage inhibitory factor. These results indicate that increasing the presence of activated macrophage/microglial cells at a spinal cord injury site can provide an environment beneficial to the promotion of regeneration of sensory axons, possibly by the release of cytokines and interaction with other nonneuronal cells in the immediate vicinity. PMID- 9417824 TI - Robust growth of chronically injured spinal cord axons induced by grafts of genetically modified NGF-secreting cells. AB - Little spontaneous regeneration of axons occurs after acute and chronic injury to the CNS. Previously we have shown that the continuous local delivery of neurotrophic factors to the acutely injured spinal cord induces robust growth of spinal and supraspinal axons. In the present study we examined whether chronically injured axons also demonstrate significant neurotrophin responsiveness. Adult rats underwent bilateral dorsal hemisection lesions that axotomize descending supraspinal pathways, including the corticospinal, rubrospinal, and cerulospinal tracts, and ascending dorsal spinal sensory projections. One to three months later, injured rats received grafts of syngenic fibroblasts genetically modified to produce nerve growth factor (NGF). Control subjects received unmodified cell grafts or cells transduced to express the reporter gene beta-galactosidase. Three to five months after grafting, animals that received NGF-secreting grafts showed dense growth of putative cerulospinal axons and primary sensory axons of the dorsolateral fasciculus into the grafted lesion site. Growth from corticospinal, raphaespinal, and local motor axons was not detected. Thus, robust growth of defined populations of supraspinal and spinal axons can be elicited in chronic stages after spinal cord injury by localized, continuous transgenic delivery of neurotrophic factors. PMID- 9417825 TI - Endogenous repair after spinal cord contusion injuries in the rat. AB - Contusion injuries of the rat thoracic spinal cord were made using a standardized device developed for the Multicenter Animal Spinal Cord Injury Study (MASCIS). Lesions of different severity were studied for signs of endogenous repair at times up to 6 weeks following injury. Contusion injuries produced a typical picture of secondary damage resulting in the destruction of the cord center and the chronic sparing of a peripheral rim of fibers which varied in amount depending upon the injury magnitude. It was noted that the cavities often developed a dense cellular matrix that became partially filled with nerve fibers and associated Schwann cells. The amount of fiber and Schwann cell ingrowth was inversely related to the severity of injury and amount of peripheral fiber sparing. The source of the ingrowing fibers was not determined, but many of them clearly originated in the dorsal roots. In addition to signs of regeneration, we noted evidence for the proliferation of cells located in the ependymal zone surrounding the central canal at early times following contusion injuries. These cells may contribute to the development of cellular trabeculae that provide a scaffolding within the lesion cavity that provides the substrates for cellular infiltration and regeneration of axons. Together, these observations suggest that the endogenous reparative response to spinal contusion injury is substantial. Understanding the regulation and restrictions on the repair processes might lead to better ways in which to encourage spontaneous recovery after CNS injury. PMID- 9417826 TI - Neuronal apoptosis and necrosis following spinal cord ischemia in the rat. AB - We examined the characteristics of neuronal death induced by ischemia in the spinal cord. Spinal cord ischemia was induced in Long-Evans rats by occlusion of the descending aorta with a 2F Fogarty catheter for 20 min (model 1) or more limited aortic occlusion (15 min) coupled with blood volume reduction (model 2); rats were sacrificed 6 h-7 days later. The animals developed variable paraparesis in model 1 and reliable paraplegia in model 2. The extent of histopathological spinal cord damage, being maximal in the lumbar cord, correlated well with the severity of paraparesis. Two distinct types of spinal cord neuronal death were observed, consistent with necrosis and apoptosis. Neuronal necrosis was seen in gray matter laminae 3-7, characterized by the rapid (6 h) onset of eosinophilia on hematoxylin/eosin-stained sections, and gradual (1-7 days) development of eosinophilic ghosting. Although TUNEL positivity was present, disintegration of membranes and cytoplasmic organelles was seen under electron microscopy. Neuronal apoptosis was seen after 1-2 days in dorsal horn laminae 1-3, characterized by both TUNEL positivity and electron microscopic appearance of nuclear chromatin aggregation and the formation of apoptotic bodies. DNA extracted from the ischemic lumbar cord showed internucleosomal fragmentation (laddering) on gel electrophoresis. These data suggest that distinct spinal cord neuronal populations may undergo necrosis and apoptosis following transient ischemic insults. PMID- 9417827 TI - Neurotrophic factors increase axonal growth after spinal cord injury and transplantation in the adult rat. AB - The capacity of CNS neurons for axonal regrowth after injury decreases as the age of the animal at time of injury increases. After spinal cord lesions at birth, there is extensive regenerative growth into and beyond a transplant of fetal spinal cord tissue placed at the injury site. After injury in the adult, however, although host corticospinal and brainstem-spinal axons project into the transplant, their distribution is restricted to within 200 micron of the host/transplant border. The aim of this study was to determine if the administration of neurotrophic factors could increase the capacity of mature CNS neurons for regrowth after injury. Spinal cord hemisection lesions were made at cervical or thoracic levels in adult rats. Transplants of E14 fetal spinal cord tissue were placed into the lesion site. The following neurotrophic factors were administered at the site of injury and transplantation: brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), neurotrophin-4 (NT-4), ciliary derived neurotrophic factor (CNTF), or vehicle alone. After 1-2 months survival, neuroanatomical tracing and immunocytochemical methods were used to examine the growth of host axons within the transplants. The neurotrophin administration led to increases in the extent of serotonergic, noradrenergic, and corticospinal axonal ingrowth within the transplants. The influence of the administration of the neurotrophins on the growth of injured CNS axons was not a generalized effect of growth factors per se, since the administration of CNTF had no effect on the growth of any of the descending CNS axons tested. These results indicate that in addition to influencing the survival of developing CNS and PNS neurons, neurotrophic factors are able to exert a neurotropic influence on injured mature CNS neurons by increasing their axonal growth within a transplant. PMID- 9417828 TI - Conduction block in acute and chronic spinal cord injury: different dose-response characteristics for reversal by 4-aminopyridine. AB - The effect of the potassium channel blocker, 4-aminopyridine (4-AP), on conduction of action potentials in injured guinea pig spinal cord axons was measured using isolated tracts in oxygenated Krebs' solution at 37 degrees C. The dose-response characteristics of acutely and chronically injured axons were compared. The maximal improvement of conduction occurred in acutely injured axons at a concentration of 100 microM 4-AP, but in chronically injured spinal cord at 10 microM. The threshold for significant response to 4-AP was between 0.5 and 1 microM in chronically injured cords, and between 1 and 10 microM following acute compression injury. The difference in susceptibility to potassium channel blockade may be related to underlying differences in the mechanism of conduction block at the two stages of injury. Initially, junctions between axons and myelin are acutely disrupted, altering primarily the leakage resistance of the myelin sheath and periaxonal space. In chronically injured cords, there is widespread but incomplete process of repair in the lesion site, which leaves many axons partially myelinated. The difference in sensitivity to 4-AP suggests there is also some modification of the accessibility of axonal potassium channel or a change in their affinity for the drug. PMID- 9417829 TI - The ability of human Schwann cell grafts to promote regeneration in the transected nude rat spinal cord. AB - Advances in the purification and expansion of Schwann cells (SCs) from adult human peripheral nerve, together with biomaterials development, have made the construction of unique grafts with defined properties possible. We have utilized PAN/PVC guidance channels to form solid human SC grafts which can be transplanted either with or without the channel. We studied the ability of grafts placed with and without channels to support regeneration and to influence functional recovery; characteristics of the graft and host/graft interface were also compared. The T9-T10 spinal cord of nude rats was resected and a graft was placed across the gap; methylprednisolone was delivered acutely to decrease secondary injury. Channels minimized the immigration of connective tissue into grafts but contributed to some necrotic tissue loss, especially in the distal spinal cord. Grafts without channels contained more myelinated axons (x = 2129 +/- 785) vs (x = 1442 +/- 514) and were larger in cross-sectional area ( x = 1.53 +/- 0.24 mm2) vs (x = 0.95 +/- 0.86 mm2). The interfaces formed between the host spinal cord and the grafts placed without channels were highly interdigitated and resembled CNS-PNS transition zones; chondroitin sulfate proteoglycans was deposited there. Whereas several neuronal populations including propriospinal, sensory, motoneuronal, and brainstem neurons regenerated into human SC grafts, only propriospinal and sensory neurons were observed to reenter the host spinal cord. Using combinations of anterograde and retrograde tracers, we observed regeneration of propriospinal neurons up to 2.6 mm beyond grafts. We estimate that 1% of the fibers that enter grafts reenter the host spinal cord by 45 days after grafting. Following retrograde tracing from the distal spinal cord, more labeled neurons were unexpectedly found in the region of the dextran amine anterograde tracer injection site where a marked inflammatory reaction had occurred. Animals with bridging grafts obtained modestly higher scores during open field [(x = 8.2 +/- 0.35) vs (x = 6.8 +/- 0.42), P = 0.02] and inclined plane testing (x = 38.6 +/- 0. 542) vs (x = 36.3 +/- 0.53), P = 0.006] than animals with similar grafts in distally capped channels. In summary, this study showed that in the nude rat given methylprednisolone in combination with human SC grafts, some regenerative growth occurred beyond the graft and a modest improvement in function was observed. PMID- 9417830 TI - Characteristics of human fetal spinal cord grafts in the adult rat spinal cord: influences of lesion and grafting conditions. AB - The present study evaluated the growth potential and differentiation of human fetal spinal cord (FSC) tissue in the injured adult rat spinal cord under different lesion and grafting conditions. Donor tissue at 6-9 weeks of gestational age was obtained through elective abortions and transplanted either immediately into acute resection (solid grafts) or into chronic contusion (suspension and solid grafts) lesions (i.e., 14-40 days after injury) in the thoracic spinal cord. The xenografts were then examined either histologically in plastic sections or immunocytochemically 1-3 months postgrafting. Intraspinal grafts in acute lesions demonstrated an 83% survival rate and developed as well circumscribed nodules that were predominantly composed of immature astrocytes. Solid-piece grafts in chronic contusion lesions exhibited a 92% survival rate and also developed as nodular masses. These grafts, however, contained many immature neurons 2 months postgrafting. Suspension grafts in chronic contusion lesions had an 85% survival rate and expanded in a nonrestrictive, diffuse pattern. These transplants demonstrated large neuronally rich areas of neural parenchyma. Extensive neuritic outgrowth could also be seen extending from these grafts into the surrounding host spinal cord. These findings show that human FSC tissue reliably survives and differentiates in both acute and chronic lesions. However, both the lesion environment and the grafting techniques can greatly influence the pattern of differentiation and degree of host-graft integration achieved. PMID- 9417831 TI - Gait analysis of adult paraplegic rats after spinal cord repair. AB - This study presents a novel detailed method of analysis of rat gait and uses this method to demonstrate recovery of forward locomotion patterns in adult rats made paraplegic by surgical spinal cord transection and subjected to a novel strategy for spinal cord repair. Six normal rats were compared to five animals in which the cord was transected at T8-T9, and a 5-mm segment of the spinal cord removed, and to seven animals in which, following spinal cord transection and removal of a spinal cord segment, multiple intercostal peripheral nerve bridges were implanted, rerouting pathways from white to gray matter in both directions. The implanted area was filled with fibrin glue containing acidic fibroblast growth factor. Details of the repair strategy have been published (H. Cheng, Y. Cao, and L. Olson, 1996, Science 273: 510-513). Gait analysis was carried out 3 and 4 months after surgery and once in the normal animals. Animals were allowed to walk across a runway with a transparent floor. Each test consisted of five trials, and each trial was videorecorded from underneath. Using frame-by-frame playback, individual footprints were then recorded regarding location and order of limb use, as well as step quality (degree of weight bearing, etc.). These data allowed measuring runway transit time, five different measures of step numbers, all possible temporal patterns of limb use, stride length, and base of support. Transected controls remained paralyzed in the hindlimbs with only occasional reflex hindlimb movements without weight bearing. Animals subjected to the full repair procedure were significantly faster than the controls, used their hindlimbs for 25-30% of the movements, and regained several of the specific limb recruitment patterns used by normal rats. Taken together, the gait analysis data demonstrate remarkable recovery of coordinated gait in the repaired animals, which was significantly better than controls for all relevant parameters, while at the same time clearly inferior to normal rats for most of the examined parameters. We conclude that normal rats use a multitude of interchangeable step sequence patterns, and that our spinal cord repair strategy leads to recovery of some of these patterns following complete spinal cord transection. These data suggest functionally relevant neuronal communication across the lesion. PMID- 9417833 TI - Axonal and nonneuronal cell responses to spinal cord injury in mice lacking glial fibrillary acidic protein. AB - We have examined the regeneration of corticospinal tract fibers and expression of various extracellular matrix (ECM) molecules and intermediate filaments [vimentin and glial fibrillary acidic protein (GFAP)] after dorsal hemisection of the spinal cord of adult GFAP-null and wild-type littermate control mice. The expression of these molecules was also examined in the uninjured spinal cord. There was no increase in axon sprouting or long distance regeneration in GFAP-/- mice compared to the wild type. In the uninjured spinal cord (i) GFAP was expressed in the wild type but not the mutant mice, while vimentin was expressed in astrocytes in the white matter of both types of mice; (ii) laminin and fibronectin immunoreactivity was localized to blood vessels and meninges; (iii) tenascin and chondroitin sulfate proteoglycan (CSPG) labeling was detected in astrocytes and the nodes of Ranvier in the white matter; and (iv) in addition, CSPG labeling which was generally less intense in the gray matter of mutant mice. Ten days after hemisection there was a large increase in vimentin+ cells at the lesion site in both groups of mice. These include astrocytes as well as meningeal cells that migrate into the wound. The center of these lesions was filled by laminin+/fibronectin+ cells. Discrete strands of tenascin-like immunoreactivity were seen in the core of the lesion and lining its walls. Marked increases in CSPG labeling was observed in the CNS parenchyma on either side of the lesion. These results indicate that the absence of GFAP in reactive astrocytes does not alter axonal sprouting or regeneration. In addition, except for CSPG, the expression of various ECM molecules appears unaltered in GFAP-/- mice. PMID- 9417832 TI - Expression of full-length trkB receptors by reactive astrocytes after chronic CNS injury. AB - Growth factors, including members of the neurotrophin family, are expressed by neuronal and glial elements following injury to the CNS. In order to assess the capacity for glial cells to respond to neurotrophins at sites of chronic injury, full-length trkB receptors were localized following implantation of a nitrocellulose filter into the cerebral cortex for 30 days. Northern analysis demonstrated that filter implants contained cells expressing transcripts for full length and truncated trkB receptors, in contrast to the predominant expression of truncated trkB receptors by cultured astrocytes. In situ hybridization and immunohistochemistry using probes to the trkB kinase domain colocalized full length receptors with GFAP-immunopositive reactive astrocytes adjacent to and within the filter implant. In contrast, OX-42-immunopositive microglia/macrophages were not stained for full-length trkB. These data indicate that reactive astrocytes can express functional trkB receptors following a chronic insult to the cerebral cortex and support the hypothesis that neurotrophins may regulate astrocytic responses to injury. PMID- 9417834 TI - FGF-2 is sufficient to isolate progenitors found in the adult mammalian spinal cord. AB - The adult rat brain contains progenitor cells that can be induced to proliferate in vitro in response to FGF-2. In the present study we explored whether similar progenitor cells can be cultured from different levels (cervical, thoracic, lumbar, and sacral) of adult rat spinal cord and whether they give rise to neurons and glia as well as spinal cord-specific neurons (e.g., motoneurons). Cervical, thoracic, lumbar, and sacral areas of adult rat spinal cord (>3 months old) were microdissected and neural progenitors were isolated and cultured in serum-free medium containing FGF-2 (20 ng/ml) through multiple passages. Although all areas generated rapidly proliferating cells, the cultures were heterogeneous in nature and cell morphology varied within a given area as well as between areas. A percentage of cells from all areas of the spinal cord differentiate into cells displaying antigenic properties of neuronal, astroglial, and oligodendroglial lineages; however, the majority of cells from all regions expressed the immature proliferating progenitor marker vimentin. In established multipassage cultures, a few large, neuron-like cells expressed immunoreactivity for p75NGFr and did not express GFAP. These cells may be motoneurons. These results demonstrate that FGF-2 is mitogenic for progenitor cells from adult rat spinal cord that have the potential to give rise to glia and neurons including motoneurons. PMID- 9417835 TI - Activated macrophages and the blood-brain barrier: inflammation after CNS injury leads to increases in putative inhibitory molecules. AB - The cellular responses to spinal cord or brain injury include the production of molecules that modulate wound healing. This study examined the upregulation of chondroitin sulfate proteoglycans, a family of molecules present in the wound healing matrix that may inhibit axon regeneration in the central nervous system (CNS) after trauma. We have demonstrated increases in these putative inhibitory molecules in brain and spinal cord injury models, and we observed a close correlation between the tissue distribution of their upregulation and the presence of inflammation and a compromised blood-brain barrier. We determined that the presence of degenerating and dying axons injured by direct trauma does not provide a sufficient signal to induce the increases in proteoglycans observed after injury. Activated macrophages, their products, or other serum components that cross a compromised blood-brain barrier may provide a stimulus for changes in extracellular matrix molecules after CNS injury. While gliosis is associated with increased levels of proteoglycans, not all reactive astrocytes are associated with augmented amounts of these extracellular matrix molecules, which suggests a heterogeneity among glial cells that exhibit a reactive phenotype. Chondroitin sulfate also demarcates developing cavities of secondary necrosis, implicating these types of boundary molecules in the protective response of the CNS to trauma. PMID- 9417837 TI - Truncated trkB receptors on nonneuronal cells inhibit BDNF-induced neurite outgrowth in vitro. AB - The function of truncated trkB receptors during nervous system plasticity and regeneration is currently unknown. The extensive nonneuronal localization of truncated trkB-T1 receptors, coupled with their up-regulation by CNS glial cells in response to injury, has led to the speculation that these receptors may sequester BDNF and NT-4/5 to reduce their local availability and, thus, limit axonal sprouting. Conversely, trkB-T1 receptors could bind and present neurotrophins to injured axons and facilitate their regeneration in a manor analogous to that proposed for p75(NTR) receptors on Schwann cells. To address this issue, we used an in vitro coculture paradigm in which wild-type 3T3 NIH fibroblasts or two different 3T3 cell clones stably expressing trkB-T1 receptors served as monolayer substrates upon which to evaluate the effect of trkB-T1 receptors on nonneuronal cells to influence neurotrophin (NGF, BDNF, NT-3, and NT 4/5)-induced neurite outgrowth from retinoic acid (RA)-treated SY5Y neuroblastoma cells. In these experiments, BDNF and NT-4/5 produce a strong phosphorylation of trk receptors on the RA-SY5Y cells and induce differentiation of the SY5Y cells (as measured by the development of neurofilament-positive neuritic processes). This ability of the trkB ligands to stimulate neurite outgrowth is dose dependent since increasing concentrations of BDNF (5, 25, and 100 ng/ml) result in an increased percentage of SY5Y cells developing neurites and in progressively longer neurites from SY5Y cells on the control 3T3 monolayers. In these experiments, BDNF and NT-4/5 induce the strongest neurite outgrowth, followed by NT-3 and then NGF. When trkB-T1 receptors are present on the 3T3 cell substratum both BDNF- and NT-4/5-induced neurite extension from the SY5Y cells are strongly inhibited. In contrast, NGF-induced neurite growth is unaffected and NT-3 associated growth is somewhat reduced. These results suggest that the inhibitory effect of the trkB-T1 receptors on the nonneuronal cell substrates is selective for neurite outgrowth that is mediated via the trkB-kinase receptors on the neuroblastoma cells. This ability of trkB-T1 receptors on the nonneuronal substratum to inhibit BDNF-induced neurite outgrowth can be overcome by the addition of high concentrations of BDNF (1 microg/ml). Binding assays using 125I BDNF suggest that this inhibitory effect could be mediated via binding and internalization of BDNF by the trkB-T1 receptors on the 3T3 cells. These results provide strong support for the hypothesis that the up-regulation of trkB-T1 receptors on astrocytes following CNS lesions enhances the sequestration of the trkB ligands, BDNF and NT- 4/5, at the site of reactive gliosis and, thus, contributes to the inhibition of CNS axonal regeneration from neurons expressing trkB-kinase receptors by removing their ligands from the extracellular environment. PMID- 9417836 TI - Schwann cells express NDF and SMDF/n-ARIA mRNAs, secrete neuregulin, and show constitutive activation of erbB3 receptors: evidence for a neuregulin autocrine loop. AB - Cultured Schwann cells secreted low levels (30 pg/ml/1.5 x 10(6) cells) of a 45 kDa neuregulin protein and showed constitutive activation of a neuregulin receptor, Erb-B3, suggesting the existence of an autocrine loop involving neuregulins in Schwann cells. RT-PCR analyses indicated that Schwann cells and fibroblasts in culture produced SMDF/n-ARIA and NDF but not GGF neuregulin messages. Schwann cell and fibroblast neuregulin messages encoded both beta and alpha domains; Schwann cell transcripts encoded only transmembrane neuregulin forms while fibroblast messages encoded transmembrane and secreted forms. SMDF/n ARIA and NDF messages were also expressed in early postnatal rat sciatic nerve, suggesting a role for neuregulins in peripheral nerve development. An anti neuregulin antibody inhibited the mitogenic response of Schwann cells to cultured neurons and to extracts of cultured neurons or embryonic brain, consistent with the accepted paracrine role of neuregulins on Schwann cells. Surprisingly, the same antibody inhibited Schwann cell proliferation stimulated by several unrelated mitogens including bFGF, HGF, and TGF-beta1. These data implicate both paracrine and autocrine pathways involving neuregulin form(s) in Schwann cell mitogenic responses. PMID- 9417838 TI - Inflammatory cytokines interact to modulate extracellular matrix and astrocytic support of neurite outgrowth. AB - Following injury to the central nervous system, an astroglial scar forms that is thought to impede neuronal regeneration and recovery of function. It is our hypothesis that inflammatory cytokines act upon astrocytes to alter their biochemical and physical properties, which may in turn be responsible for failed neuronal regeneration. We have therefore examined the interactions of two cytokines with prominent actions following injury, interferon-gamma (IFN-gamma) and basic fibroblast growth factor (FGF2), in modulating the extracellular matrix and proliferation of astrocytes in culture. We also evaluated the effects of these cytokines on the ability of astrocytes to support the growth of neurites. IFN-gamma significantly inhibited the proliferation of rat cortical astrocytes both in serum-free and serum-containing media as measured by [3H]thymidine incorporation. Furthermore, IFN-gamma also antagonized FGF2-induced proliferation. In parallel, IFN-gamma reduced the levels of the ECM molecules tenascin, laminin, and fibronectin as evaluated by Western blot analysis and immunocytochemistry. Similarly, IFN-gamma also antagonized FGF2-induced tenascin formation. While IFN-gamma-pretreated astrocyte monolayers did not differ from control in their ability to support neurite outgrowth of cortical neurons, it antagonized the enhancement of neurite outgrowth on FGF2-treated monolayers. We demonstrate that IFN-gamma did not alter signal transduction through the FGF2 receptor down to the phosphorylation of mitogen-activated protein kinase, suggesting that the interaction is at the level of transcriptional regulation or that an alternate pathway is involved. These results support the hypothesis that inflammatory cytokines interact to modulate several facets of the gliotic response and such interactions may be important in creating the biochemical and physical properties of the glial scar. PMID- 9417839 TI - Tobacco smoke habits in a group of adolescents: responsibility of the cohabitants in the active and passive exposure. AB - Several studies in adults demonstrated a positive relationship between the number of cigarettes smoked and the urinary cotinine. The aim of this work was to analyze the passive and active smoking exposure of 333 sixteen-year-old students, demonstrated by their urinary cotinine, in comparison with the smoking habits of all the cohabitants. This last information was obtained from a questionnaire and urinary cotinine in mothers. The second purpose of this work was to observe whether the smoking habits of parents can promote the voluntary active exposure to tobacco smoke in children. The considered students represent a particular group of people (16 years old) having a life-style still strictly bound to the smoking habit of parents. In fact, the nonactive smokers show urinary cotinine levels described by the following means: 28.81, 39.35, 39.62, and 57.67 ng/mL. This finding demonstrates a trend of exposure from no exposure to a maximum level of passive exposure. Similar results can be observed by considering the urinary cotinine of mothers. Finally, the active smoking habit potentially acquired by the adolescents seems to be induced also by emulation of the smoking habits of fathers. This possibility is demonstrated by a light positive and statistically significative correlation with the number of cigarettes actively smoked and, consequently, with urinary cotinine. PMID- 9417840 TI - Ventilation system, indoor air quality, and health outcomes in Parisian modern office workers. AB - A cross-sectional study was carried out to determine the effect on health of exposure to different types of ventilation, taking indoor environmental measurements (IEMs) of major contaminants and aeroallergens into account. Three buildings ventilated with heating, ventilating, and air conditioning (HVAC), fan coil units (FCUs), and natural ventilation were selected. One thousand one hundred forty-four workers answered health questionnaires. After adjusting for potential confounders, HVAC and FCU systems were related to a slightly higher risk of nonspecific symptoms (compared with natural ventilation), short-term throat irritation, work-related nasal discharge, nasal blockage on awakening, migraine, and usual coughing induced by cold air. Studying the potential effects of environmental contaminants and aeroallergens on health outcomes, taking the floor and type of ventilation into account, did not explain the observed excess of nonspecific symptoms. PMID- 9417841 TI - Cytogenetic monitoring of pesticide sprayers. AB - The induction of chromosome aberrations, micronuclei, and sister-chromatid exchanges (SCEs) was examined in cultured lymphocytes of 27 vineyard growers exposed to pesticides. Cytogenetic examinations were performed during the prespraying period, a month after spraying, and at the end of the spraying season. For comparison purposes, the same cytogenetic monitoring program was applied to two control groups. The first consisted of 15 individuals from a nearby town, and the second consisted of 20 volunteers living 200 km from the vine-growing area (reference control group). A positive, though low statistically significant (P < 0.017) difference in the yield of unstable chromosomal aberrations in exposed sprayers was observed compared with both control groups during the prespraying period. The mean group value of micronuclei in exposed workers averaged 5.41 per 1000 binucleated cells, with individual means ranging from 0 to 15. In both control groups, the yield of micronuclei averaged 5.09 per 1000 binucleated cells, with individual means ranging from 1 to 10. No statistically significant (P < 0.5) differences in yield of micronuclei were found in exposed subjects compared with both control groups. Significant individual variation (F = 14.09, P < 0.000) in SCE frequency was observed in exposed subjects, as well as in both control groups (F = 14.09, P < 0.000). A month after spraying, the average incidence of unstable aberrations in pesticide sprayers was 0.22%, and the yield of micronuclei averaged 17.78 per 1000 binucleated cells, with individual means ranging from 7 to 28. The incidence of micronuclei a month after spraying in exposed subjects was elevated (statistically significant at P < 0.01) in comparison with the prespraying period, while the difference in the yield of chromosomal aberrations in exposed subjects was insignificant (P < 0.5). At the end of the spraying season, the average incidence of unstable aberrations in exposed subjects was 0.79%, and the yield of micronuclei averaged 39.92 micronuclei per 1000 binucleated cells, with individual means ranging from 21 to 62. The appearance of more than one micronucleus per binucleated cell was related to the results on chromosome aberrations. The frequencies of chromosomal aberrations and micronuclei were significantly higher (P < 0.001, P < 0.000) in the exposed group than in their matched control groups. The yield of micronuclei in pesticide sprayers at the end of the season was higher than expected with respect to chromosomal aberration frequency, which provides some evidence that some of the micronuclei are induced by the spindle-inhibiting effects of pesticides. A statistically significant (P < 0.003) difference in micronuclei in the first control group was observed compared with the reference control group at the end of the spraying season. With respect to the incidence of micronuclei in the control group in the vine-growing area, a poor but positive correlation (r = 0.074, P < 0.104) with duration of the spraying season was found, which is probably due to airborne pesticides in the vine-growing area. SCE frequencies of the workers' lymphocytes were not significantly changed due to the exposure. The yield of aberrations as well as that of micronuclei in exposed subjects correlated positively (r = 16, P = 0.016) with duration of exposure. PMID- 9417842 TI - Effects of Aroclor 1254 on the thyroid gland, immune function, and hepatic cytochrome P450 activity in mallards. AB - Adult male mallards were exposed to 0, 4, 20, 100, 250, and 500 mg/kg Aroclor 1254 by gavage twice per week for 5 weeks. Immunotoxic effects, as measured by antibody titers to sheep erythrocytes, natural killer cell activity and lymphocyte mitogenesis to phytohemagglutinin, were not detected as a consequence of polychlorinated biphenyl (PCB) exposure. Hepatic cytochrome P450 activities were measured as microsomal dealkylations of ethoxyresorufin (EROD) and pentoxyresorufin (PROD). Significant elevations in EROD and PROD were noted at 20 mg/kg and peaked in birds treated with 100 mg/kg. Total P450 was induced beginning at 100 mg/kg and peaked at 250 mg/kg. Relative liver weights were dose dependently increased following treatment with 100 mg/kg or more. Thyroid weights were significantly increased in PCB-treated birds treated with 100 mg/kg or greater, but no significant histological abnormalities were observed, except at the highest dose. Plasma total triiodothyronine (T3) was decreased in a dose dependent manner, with a significant lowest-observed-adverse-effect level (LOAEL) of 20 mg/kg. T3 was decreased following 7 days treatment with 100 mg/kg. The no observed-adverse-effect level (NOAEL) was 4 mg/kg for decreased T3. Plasma glucose levels were decreased on days 28 and 35 in mallards treated with 500 mg/kg, while other clinical plasma biochemistry parameters were unaltered by PCB treatment. Plasma corticosterone levels were unchanged by PCB treatment. These results indicate that thyroid hormone levels and P450 activity in mallards are sensitive to subchronic PCB exposure in the absence of gross toxic effects and immunotoxicity. PMID- 9417843 TI - Estrogenic effects of p-hydroxybenzoic acid in CD1 mice. AB - Xenobiotic estrogens in the environment or diet have received much attention as a possible source of certain hormonal disease states in human and wildlife. Therefore, the detection of estrogenic activity of any substance, especially those related to the food industry, is important. The estrogenic activity of p hydroxybenzoic acid (PHBA), a compound related to a commonly used group of preservatives in food, cosmetic, and pharmaceutical preparations, was evaluated with immature and adult ovariectomized female mice (CD1) using two well-known bioassays. Subcutaneous administrations (s.c.) of different doses of PHBA were compared with estradiol (E2), and their effects on vaginal cornification and uterotrophic activities were evaluated. Different groups of animals were treated s.c. daily for 3 days with vehicle (corn oil, 0.3 ml/100 g), E2 (1 microgram/100 g), and PHBA (0.5, 5, 50, and 500 micrograms/100 g). Four days after treatment, PHBA produced a dose-dependent response on vaginal cornification and uterotrophic activity in both immature and adult ovariectomized mice. The relative uterotrophic potency of PHBA (500 micrograms/100 g) to E2 (1 microgram/100 g) was 0.0011 in immature mice and 0.0018 in ovariectomized animals. PMID- 9417844 TI - Cellular mechanisms of reactive oxygen metabolite generation from human polymorphonuclear leukocytes induced by crocidolite asbestos. AB - In our previous study, we demonstrated that reactive oxygen metabolites (ROM) generation from phagocytic cells may be involved in the carcinogenic mechanism of crocidolite asbestos. In the present study, the mechanism of human polymorphonuclear leukocytes (PMN) to generate ROM by crocidolite was investigated using verapamil, a calcium channel inhibitor; staurosporine, a NADPH oxidase inhibitor; and cytochalasin B (CB), an inhibitor of phagocytosis. The results indicate that whereas verapamil and staurosporine inhibited the crocidolite-induced ROM generation from PMN dose-dependently, CB caused an enhancement. We conclude that crocidolite-induced ROM generation involves a cell surface reaction due to influx of extracellular calcium through calcium channels and the activation of NADPH oxidase on the PMN cell membrane. This hypothesis was indirectly supported by dose-dependent enhancement of the ROM generation by CB, as CB increases calcium ion permeability in PMN. However, as in our previous studies, the time course of the ROM generation and the cell type difference suggested that ROM were also generated intracellularly from PMN due to phagocytosis of crocidolite. In conclusion, our evidence indicates that ROM generation from PMN by crocidolite involves cellular mechanisms related both to direct cell surface membrane interactions, together with an apparent phagocytic dependent process. PMID- 9417845 TI - Reaction of human alveolar macrophages to exposure to Aspergillus fumigatus and inert particles. AB - In vitro interaction of human alveolar macrophages (AM) with heat-killed conidia from Aspergillus fumigatus and inert silica particles of similar size, about 3 microns, was studied. The conidia were phagocytized significantly faster by AM than were the control particles partly due to the faster rate of attachment but especially due to the faster rate of ingestion. Quantitative nitroblue tetrazolium (NBT) reduction by AM, reflecting their release of oxygen radicals, was increased by a factor of 2 to 3 in response to the conidia during phagocytosis. The silica particles induced a moderate but significant increase in NBT reduction. Conidia, but not silica particles, showed a considerable percentage (around 8%) of phagolysosomes with neutral pH after 3 h and a smaller percentage (around 1%) after 24 h of incubation. The pH of phagolysosomes with conidia tended to be higher after 3 h, but was significantly lower after 24 h than the pH of phagolysosomes with silica particles. Despite the markedly increased oxidative metabolism there was no increase in cytokine production [interleukins (IL) 6 and 8 and tumor necrosis factor alpha (TNF-alpha)] after exposure to conidia. The silica particles induced a significant decrease in IL-6 and IL-8 production and a tendency toward decreased production of TNF-alpha. The occurrence of phagolysosomes with neutral pH suggests unsealed phagolysosomes from which not only oxygen metabolites but also enzymes might escape from the cell. Lung damage may thus be the result of repeated or long-term exposure to Aspergillus conidia. PMID- 9417846 TI - Ambient air levels and the exposure of children to benzene, toluene, and xylenes in Denmark. AB - The aims of the study were to evaluate if the front-door concentrations of benzene, toluene, and xylenes can be used to classify the personal exposures of Danish children and to identify factors that affect their personal exposure. Average concentrations were measured over 1 week with diffusive samplers, and the personal exposures of 98 children and the concentrations outside the front doors of their homes were measured simultaneously. Time and activity patterns were noted in diaries. The front-door concentrations were significantly higher in Copenhagen than in rural areas (all P < 0.0001), but the personal exposures were only slightly higher. Even though the personal exposures were highly significantly associated with front-door concentrations in urban areas (all P < 0.004), use of the residential front-door concentration as an exposure surrogate would imply misclassification, as it cannot be used for rural children. Multiple regression analyses brought to light several factors that affect the exposure of children independently, including front-door concentration, riding in cars, and activities involving potential exposure to gasoline vapors like motocross, moped driving, and refueling of cars. PMID- 9417847 TI - Risk, mercury levels, and birds: relating adverse laboratory effects to field biomonitoring. AB - There is an abundance of field data on levels of mercury in a variety of organisms and there are a number of studies that demonstrate the adverse effects of mercury on laboratory animals, but few studies examine the relationship between the two. Thus it is often difficult to determine the ecological relevance of mercury concentrations found in nature, or to predict the ecosystem consequences of current levels. In this paper we review the levels in tissues that are associated with adverse effects in birds from laboratory studies and compare these with levels found in wild bird populations in the New York Bight to provide a basis for interpreting values in avian populations. We use feathers from fledgling birds which would have been fed on locally obtained food to eliminate the problem of where toxic burdens were acquired by more mobile adult birds. Laboratory studies indicate that in some species mercury levels of 1.5 ppm in eggs and/or 5 to 40 ppm in the feathers of birds are associated with adverse effects, including impaired reproduction. We report egg levels in birds that range as high as 3.8 ppm and feather levels that range as high as 10.3 ppm, although means are much lower. The levels in eggs of some wild birds in the New York Bight are within the range known to lower hatchability, embryo and chick survival, and chick weight, all variables that reduce reproductive success. Species with high egg levels include Forster's tern (Sterna forsteri) and black skimmer (Rynchops niger). Levels in feathers of some young wild birds from the New York Bight are within the range associated with reduced hatchability of eggs, behavioral abnormalities of adults, and infertility. Species with dangerously elevated mercury levels in feathers include great egret (Ardea [=Egretta] alba), snowy egret [Egretta thula), and black skimmers. PMID- 9417848 TI - Metal levels in mourning doves from South Carolina: potential hazards to doves and hunters. AB - Most game birds are found in lower trophic levels, but since such birds are harvested and consumed by humans, there is a particular need to assess their contaminant levels. In this paper, we report concentrations of mercury, lead, cadmium, selenium, manganese, and chromium in the breast feathers, liver, and muscle of mourning doves (Zenaida macroura) collected at a partially drawn-down, contaminated reactor-cooling reservoir (Par Pond) on the Department of Energy's Savannah River Site in South Carolina and at nearby agricultural fields managed as dove hunting areas. We test the hypothesis that the levels in doves are not harmful to either dove populations or humans. We also tested the simultaneous effects of collection location, year (1992, 1993), and dove age-class (hatch-year vs after hatch-year) on heavy metal and selenium levels. For all three tissues, mercury levels were nondetectable at all locations. Lead was highest in tissues from agricultural fields with prior histories of dove hunting activities. Doves at those fields were likely ingesting lead shot to a greater degree than at the recently drawn-down reservoir which was closed to public access and hunting. For other metals, Par Pond doves had equally high or higher tissue levels. For all metals, levels in doves from South Carolina were generally within the lower range of those reported in the literature, suggesting that these metals were likely to pose no health problems to these doves. Except for lead and selenium, metal levels in dove muscle that we observed were well below reference metal doses established for human intake. Lead and selenium, at the levels described here, would only be a problem if a child (not an adult) ate 120 g of dove meat every day of the year. Thus, we conclude that meat from these doves, if consumed by hunters, would not pose a risk. PMID- 9417849 TI - Comparison of the bioaccumulation capacities of copper and zinc in two snail subspecies (Helix). AB - Bioaccumulation analyses of copper and zinc were carried out in two snail subspecies (Helix aspersa aspersa and Helix aspersa maxima) after 3 months of controlled farming (out of ground) with foods of different formulations. The results reveal some clear interspecific differences in affinity toward copper and zinc. For the two metals considered, H. aspersa aspersa has a bioaccumulation capacity much greater than that of H. aspersa maxima, mainly in the foot for copper and in the viscera for zinc. After 3 months, the concentrations of copper in feet and viscera are much higher than those presented in the literature on field animals. The farming and the analysis methodologies permitted obtaining snails under standard condition and open the way to the development of rational protocols for ecotoxicological studies in a laboratory as well as in the field. PMID- 9417850 TI - Effects of methyl mercury on the in vivo release of dopamine and its acidic metabolites DOPAC and HVA from striatum of rats. AB - Mercury is a neurotoxic agent that produces different effects on the brain. In the present work, the effects of chronic doses of methyl mercury (MeHg) were studied on the dopaminergic system of the rat striatum, using microdialysis coupled to high-performance liquid chromatography in order to quantify the in vivo release of dopamine (DA) and its acidic metabolites DOPAC and HVA. The administration of an equivalent total dose of 6 mg/kg of MeHg induced significant increases in the striatal release of DA and/or its acidic metabolites, independently of the pattern of administration. These effects are discussed on the base of the release and the metabolization of DA. In conclusion, the effect of MeHg administered under these experimental conditions on the in vivo release of DA and its metabolites seems to have a dose-dependent component and seems to be an accumulative process. PMID- 9417851 TI - Effects of an acute exposure to lindane (gamma-hexachlorocyclohexane) on brain and liver carbohydrate metabolism of rainbow trout. AB - The capacity of carbohydrate and ketone bodies metabolism in brain and liver was evaluated in rainbow trout control and exposed to lindane (0.05 mg.L-1) for 12 hr. The results obtained demonstrate the existence of changes in several parameters of brain carbohydrate metabolism due to lindane treatment. Thus, increased plasma glucose levels are reflected in brain by the mobilization of glycogen stores and increased lactate levels probably reflecting an increased anaerobic use. Also, ketone bodies appear to be used under this stressful condition. In liver, the main results obtained suggest that glycogen stores are being mobilized to be probably used as glucose in pathways other than glycolysis. PMID- 9417852 TI - Single and joint toxic effects of copper and zinc on reproduction of Enchytraeus crypticus in relation to sorption of metals in soils. AB - Joint toxic effects of copper and zinc were studied in the terrestrial worm Enchytraeus crypticus (Westheide and Graefe) (Oligochaeta, Annelida). Animals were exposed in OECD artificial soil. Sublethal toxicity was judged by effects on reproduction. Metals were applied singly or in binary mixtures. Observed effects were compared with effects expected from simple similar action (concentration addition), by recalculation of metal concentrations in toxic units. Exposure of the worms was quantified with body concentrations and with external concentrations (total, extractable, soluble). The observed joint effect was similar to concentration additive when judged by external concentrations and less than concentration additive for body concentrations. This difference is attributable to interactions among metals during sorption to soil and during uptake. Copper reduced the sorption of zinc to soil, but copper sorption was inert for zinc addition. Zinc uptake from the soil solution was stimulated by copper, but copper uptake was not stimulated by zinc. Joint effects of toxicants to soil biota are partly determined by interactions outside the organism, as a result of dissimilarity between total and bioavailable concentrations. The design of joint toxicity studies in terrestrial systems is discussed with special reference to metal sorption in soils, experimental methodology, and laboratory practice. The joint toxic effect of copper and zinc for E. crypticus was of similar magnitude as found in studies with aquatic species exposed to metal mixtures. PMID- 9417853 TI - Glutathione reductase, selenium-dependent glutathione peroxidase, glutathione levels, and lipid peroxidation in freshwater bivalves, Unio tumidus, as biomarkers of aquatic contamination in field studies. AB - The aim of this study was to evaluate the usefulness of antioxidant parameters in the freshwater bivalve, Unio tumidus, as biomarkers of exposure to pollutants and to study their potential interest in predicting toxicity. Selenium-dependent glutathione peroxidase (Se-GPx), non-selenium-dependent glutathione peroxidase (non-Se-GPx), glutathione reductase (GRd), catalase, and superoxide dismutase (SOD) activities; reduced (GSH) and oxidized (GSSG) glutathione levels; and lipid peroxidation were measured in the gills and digestive glands of Unio. Control mussels were encaged and transplanted for 15 and 30 days to sites where the contamination of sediments was analyzed, along a river receiving domestic and industrial sources of pollution. After 15 days of exposure, all antioxidant parameters of the bivalves transferred to the most polluted sites had strongly decreased compared with control values. This was particularly true for Se-GPx and GRd activities, which were inhibited by 60 and 80% in the two tissues, and for GSH levels (80% reduction in the gills and 60% in digestive glands). These decreases were associated in the gills with lipid peroxidation (measured by malondialdehyde content) and with a high level of contamination of sediments by polycyclic aromatic hydrocarbons and polychlorinated biphenyls. In the mussels exposed at the least polluted sites, the same parameters decreased in the gills, but to a lesser extent: 50% for Se-GPx and 32% for GRd activities, and 45% for GSH levels. The gills appeared more sensitive than the digestive glands. After 30 days of exposure, while Se-GPx, GRd, and GSH remained reduced, a significant induction of non-Se-GPx and catalase activities was recorded in the gills, which reflected an adaptation of the transplanted species to their unsafe environment. All the results indicated that antioxidant defense components, namely, Se-GPx, GRd, and GSH, are sensitive parameters that could be useful biomarkers for the evaluation of contaminated aquatic ecosystems. The relationship between the degree of deficiency of antioxidant defenses and lipid peroxidation suggests that these parameters could also be biomarkers for toxicity. PMID- 9417854 TI - Inhibition of gill Na+,K+-ATPase activity in the eel, Anguilla anguilla, by fenitrothion. AB - European eels (Anguilla anguilla) were exposed to sublethal fenitrothion concentrations (0.02 and 0.04 mg/liter) in a continuous flow-through system for 4 days. Gill Mg2+- and Na+,K+-ATPase activities were evaluated after 2, 8, 12, 24, 32, 48, 56, 72, and 96 h of pesticide exposure. Results indicated that ATPase activity in gill tissue decreased as concentration of fenitrothion increased. Pesticide induced significant inhibitory effects on the Na+, K+-ATPase activity of A. anguilla, ranging from >56% inhibition at a sublethal concentration of 0.02 ppm to >73% inhibition at a sublethal concentration of 0.04 ppm. Eels were exposed to both fenitrothion concentrations for 96 h and then allowed a period of recovery in pesticide-free water. Samples were removed at 8, 12, 24, 48, 72, 96, 144, and 192 h and eel gill ATPase activity was evaluated. Following 1 week of recovery, the Na+,K+-ATPase activity for those animals previously exposed to 0.04 ppm fenitrothion was still different from that of the controls. PMID- 9417855 TI - Cadmium bioaccumulation in carp (Cyprinus carpio) tissues during long-term high exposure: analysis by inductively coupled plasma-mass spectrometry. AB - This work was an attempt to investigate cadmium bioaccumulation in the carp (Cyprinus carpio) during simulated pollution. The fish, weighing 100 g, were kept for 140 days in a 1000-liter indoor concrete tank supplied with a continuous flow (8 liters min-1) of unchlorinated, aerated, and filtered well water, the cadmium concentration of which was maintained at 450 microg liter-1. Such a high cadmium concentration was chosen to achieve metal saturation of the fish organs. Carp were fed during exposure. Cadmium accumulation in liver, kidney, and muscle was measured by inductively coupled plasma-mass spectrometry (ICP-MS), which is one of the most sensitive analytical techniques. The reported data indicate that cadmium exposure results in significant cadmium uptake, but the pattern of this uptake varies with the organ. For kidney and liver, cadmium concentration increased rapidly to the saturation level, probably because of the limited ability to store the cadmium as exposure persisted. A positive correlation between the increase in metallothioneins in tissues and the increase in metal tolerance in fish has been suggested. The cadmium concentration increase in muscle was significant only after 3 months, and, then, increased as an exponential function of the exposure time. When the storage capacity limits of the liver and kidney are reached, cadmium accumulation in muscle is stimulated. In this experiment, the high contamination levels reached suggest that such muscle would be unsuitable for human consumption. After 140 days of exposure, the cadmium concentrations in muscle, liver, and kidney were respectively 9 +/- 1, 91 +/- 7, and 250 +/- 16 mg kg-1 dry wt. PMID- 9417856 TI - Effects of the herbicide molinate on mixotrophic growth, photosynthetic pigments, and protein content of Anabaena sphaerica under different light conditions. AB - The effects of the carbamate herbicide molinate on growth and chlorophyll a, biliprotein, and protein content of the nitrogen-fixing cyanobacterium Anabaena sphaerica grown mixotrophically under 3000- and 300-lux light intensity were studied. Under two light intensities, the three concentrations of molinate tested (5, 25, and 50 microg ml-1) can significantly inhibit algal growth in a dose dependent manner. The high concentration of molinate (50 microg ml-1) stimulated the synthesis of chlorophyll a and protein, but inhibited the formation of biliprotein, and these effects appear to be greater at 3000 lux than at 300 lux. The effects of the other two concentrations of molinate (5 and 25 microg ml-1) on chlorophyll a and biliprotein varied with time, but 5 microg ml-1 molinate had an inhibitory effect on the synthesis of protein. It was demonstrated that different concentrations of molinate had different effects on chlorophyll a, biliprotein, and protein content, which varied with light intensity. The results support the suggestion that the physiological mode of molinate toxicity to A. sphaerica is related to its interference with the metabolism of protein, particularly the formation and functional effectiveness of some special protein. PMID- 9417857 TI - Lethality of pyrethrins to larvae and postlarvae of the American lobster (Homarus americanus). AB - Pesticide formulations containing pyrethrins are being used to treat salmonids for infestations of the copepod parasites Lepeophtherius salmonis and Caligus elongatus (sea lice). The acute lethality of one such formulation to four larval stages of the American lobster (Homarus americanus), a species of significant economic importance in eastern Canada, was determined. The formulation tested contained 0.06% pyrethrins and 0.6% piperonyl butoxide (a synergist). Stage I larvae (48-h LC50 = 4.42 microg/liter) were significantly less sensitive than stage II, III, or IV larvae. Stage II larvae (48-h LC50 = 2.72 microg/liter) were significantly less sensitive than Stage III or IV larvae. Stage III and IV larvae were not significantly different in their response to the pyrethrins formulation (48-h LC50 = 1.39 and 0.73 microg/liter, respectively). Most published studies using lobster larvae have reported that the earliest larval stage was the most sensitive to chemicals. The results described here indicate that the earliest larval stage is the least sensitive to the pyrethrins formulation. PMID- 9417858 TI - Toxicity of 4-nonylphenol in a life-cycle test with the midge Chironomus tentans. AB - A life-cycle test with the macroinvertebrate Chironomus tentans was conducted with 4-nonylphenol. The chironomids were exposed to a series of concentrations of 4-nonylphenol via the water, in an intermittent (2 times/day) water renewal system. The test included evaluation of a number of developmental (e.g., growth) and reproductive (e.g., emergence, fecundity, viability) endpoints through parental and into F1 generations. Reductions in survival were observed in 20-day old larvae at the highest test concentration, which corresponded to no-observable and lowest-observable-effect concentrations of 42 and 91 microg/liter, respectively. No significant effects on larval growth (20 days), organism survival past 20 days, emergence success or pattern, sex ratio, fecundity, or egg viability were observed at any treatment level. Qualitative observations indicated an increase in deformed egg masses at the highest test concentrations; however, the biological significance of this is uncertain. PMID- 9417859 TI - The retinoblastoma protein pathway in cell cycle control and cancer. AB - Recent discoveries in diverse fields of biomedical research have merged to reveal the molecular basis of cell cycle control and a critical role of subverting this homeostatic mechanism in cancer development. At the heart of these processes lies a late G1 checkpoint governed by the "RB pathway" whose molecular composition, functions, and cancer-associated defects are briefly evaluated in this review. This exciting new knowledge raises a plethora of conceptual issues in cell biology, with potential practical implications for biotechnology and medicine. PMID- 9417860 TI - The tumor suppressor protein p16INK4a. AB - The tumor suppressor protein p16INK4a (inhibitor of CDK4) is one of the most direct links between cell-cycle control and cancer. The p16INK4a gene is frequently inactivated in human tumors, and inheritance of mutant alleles results in susceptibility to several types of cancer. p16INK4a is part of a cell-cycle regulatory pathway that converges in the tumor suppressor protein Rb. The mechanisms that regulate p16INK4a are starting to be characterized. PMID- 9417861 TI - p53: emergency brake and target for cancer therapy. PMID- 9417862 TI - Alpha-melanocyte-stimulating hormone and endothelin-1 have opposing effects on melanocyte adhesion, migration, and pp125FAK phosphorylation. AB - Recent reports show that alpha-MSH (melanocyte-stimulating hormone) is mitogenic and melanogenic for normal human melanocytes, and that this effect is mediated through binding to the melanocortin receptor (MC1R) and activation of cAMP formation. alpha-MSH has also been shown to induce changes in cell shape in melanocytes and melanoma cells, particularly increased dendricity, suggesting a potential role for alpha-MSH in melanocyte-matrix interactions and pigment transfer through reorganization of the melanocyte actin filament cytoskeleton. In this report we show that the potent alpha-MSH analog (Nle4, D-Phe7)-alpha-MSH (NDP-MSH) induces reorganization of the actin stress fiber cytoskeleton in treated human melanocytes and that this reorganization is associated with increased adhesion to fibronectin (FN). Because most melanocyte growth factors act synergistically on melanocyte mitogenesis, we also sought to determine the effect of the melanocyte mitogen endothelin-1 (ET-1) on the melanocyte actin cytoskeleton, melanocyte adhesion, and melanocyte migration. We show that ET-1, which increases melanocyte migration on FN, has opposite effects on melanocyte adhesion to FN compared with NDP-MSH and that endothelin-1-induced actin reorganization is distinct from that observed following NDP-MSH treatment. Finally, we show that focal adhesion kinase (pp125FAK), a nonreceptor tyrosine kinase associated with focal contact formation and cell migration, is phosphorylated on tyrosine residues after treatment of melanocytes with ET-1, but not NDP-MSH. These data indicate that while alpha-MSH and ET-1 act synergistically to modulate melanocyte proliferation, they have opposite effects on melanocyte-matrix interactions. PMID- 9417863 TI - Geldanamycin prevents nuclear translocation of mutant p53. AB - p53 is a tumor suppressor protein that acts in the nucleus to effect cell cycle arrest and apoptosis. In some cells p53 is located in the cytoplasm, perhaps as a means of downregulating its activity. We recently showed that hsp90 forms a complex with the cytoplasmically localized mutant p53 (TSp53vall35) within transformed cells (Sepehrnia et al., J. Biol. Chem. 271, 15084, 1996). The present study was undertaken to determine the p53 conformation bound to hsp90 and the role of hsp90 in p53 nuclear translocation. We show that hsp90 binds both a native and a denatured form of p53 as determined by conformation-specific antibodies. hsp90 does not bind p53 in a spatial-specific manner because it remains bound to p53 when induced to translocate to the nucleus by the protein synthesis inhibitor cycloheximide (CHX). Treatment of transformed cells with geldanamycin (GA), a small molecule that binds hsp90, causes a rapid destabilization of p53 by 50%. Residual p53 that survives GA treatment is incapable of translocating to the nucleus. GA does not destabilize p53 in cells where p53 is genotypically wild type. Although GA appears to dramatically alter the translocating properties of mutant p53 it does not dissociate the p53-hsp90 complex. We suggest that a second chaperone protein, called p23, which we show also binds p53, may play an important role in these GA-mediated effects. PMID- 9417864 TI - Elevated levels of mortalin expression in human brain tumors. AB - We have performed immunohistochemical studies of mortalin in normal and tumor human brain sections. In normal brain sections, the expression was seen mainly as being confined to neurons. Normal astrocytes showed undetectable expression of this unique member of the heat shock 70 protein family. Three grades of astrocyte tumors (low-grade astrocytoma, anaplastic astrocytoma, and glioblastoma), however, showed an increasing number of mortalin-positive cells. Other types of brain tumors, such as meningiomas, neurinomas, pituitary adenomas, and metastases, also showed elevated levels of mortalin expression compared to those in the normal brain. Mortalin has earlier been reported to have differential intracellular distribution in normal and transformed cells in vitro. Therefore, we substantiated the present study with immunofluorescence localization of the protein in normal and glioblastoma cells. The observations indicated that the tumors might be expressing a nonpancytosolic mortalin. An increase in number of mortalin-positive cells with malignant progression of brain tumors and its correlation with Ki-67 (a cell proliferation marker)-positive cells further suggested an involvement of nonpancytosolic mortalin(s) in malignant transformation of cells in vivo. PMID- 9417865 TI - Induction and expression of human cartilage glycoprotein 39 in rheumatoid inflammatory and peripheral blood monocyte-derived macrophages. AB - Human cartilage glycoprotein 39 (HC gp-39) has been described as a major secreted product of cultured articular chondrocytes, synovial fibroblasts, and the osteosarcoma line MG63. However, its expression in these cells types has not been directly linked to corresponding cell types in vivo. In this report, expression of HC gp-39 is demonstrated from peripheral blood-derived macrophages in association with their differentiation from monocytes to macrophages. Consistent with macrophage specificity, HC gp-39 expression is also induced upon selective stimulation of the pluripotent promyelocytic leukemia cell line HL-60 toward the monocyte/macrophage lineage with vitamin D3 or phorbol 12-myristate 13-acetate (PMA), while treatments stimulating granulocyte and eosinophilic pathways do not induce expression. Furthermore, HC gp-39 expression levels correlate with the degree of morphological differentiation induced by PMA and vitamin D3 treatments. PMA-induced mRNA expression occurs by 36 h and is a secondary transcriptional response since its synthesis is inhibited by cycloheximide. Apparently, HC gp-39 expression is tied to later events in the differentiation of monocytes into macrophages. The in vivo significance of these results is validated by the in situ detection of HC gp-39 mRNA in inflammatory macrophages associated with rheumatoid synovium. Thus, macrophages appear to be an important source of HC gp 39, which has been shown to be present at elevated levels in the blood and synovium of rheumatoid arthritis patients. The implications of this extend well beyond the previously restricted observations in cell types associated with the joint and suggest a potential involvement of macrophage-derived HC gp-39 in other aspects of inflammation, tissue remodeling, and host defense. PMID- 9417866 TI - Mouse hepatitis virus infection induces an early, transient calcium influx in mouse astrocytoma cells. AB - Mouse hepatitis virus (MHV), a murine coronavirus, utilizes murine carcinoembryonic antigens as receptors. The events that follow virus-receptor binding and eventually lead to virus entry are poorly understood. We studied the possible effects of MHV infection on intracellular calcium in a mouse astrocytoma cell line. Using the calcium-sensitive dye fluo-3 and confocal laser scanning microscopy, we found that MHV strain JHM induced an immediate (within 20 s) and transient (lasting no longer than 2 min) calcium increase in about 5% of the infected cells. The calcium increase was blocked by antibodies against the viral spike protein, suggesting that it was specifically triggered by the interaction of the viral spikes with cells. It was also inhibited by L-type calcium channel blockers and was not detected in calcium-free medium, suggesting that the calcium increase was caused by calcium influx from the extracellular medium. Studies of the kinetics of viral replication by immunofluorescence staining of the viral nucleocapsid protein revealed that at 3 h postinfection there was roughly the same percentage of cells (5%) that produced the viral protein as the percentage of cells that had responded with a calcium signal. This finding and the virus dilution studies together suggest that calcium responders may represent cells that had been infected with multiple viruses and undergone rapid viral replication. Furthermore, calcium channel blockers, including verapamil and cadmium chloride, and the calcium chelator EGTA inhibited virus infection. Therefore, the transient intracellular calcium increase reported here may be an early signaling event associated with virus infection. PMID- 9417867 TI - Multiple protein: protein interactions between the snRNP common core proteins. AB - The snRNP core proteins (B, D3, D2, D1, E, F, and G) assemble with snRNA and form the snRNP core particle with a suggested stoichiometry of B2[D1, D2(E, F, G)2]D3. The newly synthesized snRNP core proteins are stored in the cytoplasm in three RNA-free complexes of (1) B at 2S-6S; (2) [D1, D2(E, F, G)2] at 6S; and (3) (B, D3, and 69 kDa) at 20S. The snRNP proteins assemble stepwise with snRNAs that appear transiently in the cytoplasm before returning to the nucleus as mature snRNP particles. In this report, two approaches are used to investigate the protein:protein interactions between the snRNP proteins. First, the 6S and 20S cytoplasmic complexes chromatographed as intact structures, supporting their identifications as discrete complexes. Second, the cDNAs for the proteins were used to test all pair-wise interactions between the seven major core proteins using the yeast two-hybrid system. The two-hybrid system identified four strong reciprocal interactions, one weak reciprocal interaction, five one-way interactions, and one homotypic interaction. The strongest interactions were between proteins within the 6S particle. Other interactions were between proteins in the 6S and 20S particles or within the 20S particle itself. These interactions are likely to occur within the cytoplasmic snRNP core protein complexes and the mature snRNP particle. PMID- 9417868 TI - Laminin alpha 5, a major transcript of normal and malignant rat liver epithelial cells, is differentially expressed in developing and adult liver. AB - The laminin family of extracellular matrix glycoproteins plays a major role in cell migration and differentiation and in tumor cell invasion. As previously shown, the laminin deposited by normal and malignant rat liver epithelial cells in their extracellular matrix (ECM) and into their ECM migration tracks does not contain a typical (EHS-like) alpha 1 heavy chain. By RT-PCR screening we have now identified two alpha chains among a total of five additional laminin chains produced by these cells. Three of the newly identified chains were not previously known for the rat. Their sequences have been deposited in the EMBL nucleotide sequence data bank. The alpha 5 chain now identified is expressed at comparably high levels by both the normal and the malignant liver epithelial cells. The chain is also expressed in fetal liver together with the alpha 2 and beta 2 chains, but it is only vestigially expressed in the mature organ as shown by RT PCR. These results suggest for alpha 5 a role in development and production of the chain by only a small subset of cells in adult liver. At the level of detection used, no changes were observed in regenerating liver after partial hepatectomy. In addition to the alpha 5 chain, the cultured cells express the beta 1 and beta 2 light chains, indicating the expression of more than one laminin isoform by the same cell line. The expression of the alpha 5 chain and of the other new non-EHS isoform chains was also analyzed in various tissues. The malignant liver epithelial cells, but not their nontumorigenic parental cells, also express, in addition to the alpha 5 chain the alpha 2 chain, which is expressed at high level by the NBT II bladder carcinoma cell line, suggesting a relationship with malignancy. PMID- 9417869 TI - Aging reduces the numbers of hepatocytes synthesizing DNA in response to EGF and epinephrine. AB - Primary cultures of hepatocytes were prepared from young (6 month) and old (24 month) Wistar rats and exposed to epinephrine or epidermal growth factor. Incorporation of [3H]thymidine into DNA was determined both radiochemically and autoradiographically. The numbers of responding cells and degree of response per cell were determined and the results confirmed by FACScan analysis. Such analyses clearly demonstrate a reduced number of hepatocytes capable of responding to the above stimuli in cultures obtained from old rats. Thus, changes in numbers of responding cells may be an important mechanism involved in reduced responsiveness of the aged liver to agents which stimulate DNA synthesis and cell division. PMID- 9417870 TI - Characterization of density-dependent regulation of the tyrosinase gene promoter: role of protein kinase C. AB - The rate-limiting step in melanogenesis is catalyzed by tyrosinase, a multifunctional enzyme encoded by the albino locus. We have previously reported that depletion of protein kinase C by long-term treatment of B16 mouse melanoma cells with phorbol dibutyrate (PDBu) prevented cell density-dependent melanogenesis. This was accompanied by a lack of induction of tyrosinase protein and mRNA. We report here the effect of PDBu on the functional activity of the mouse tyrosinase promoter by reporter gene assay and its effect on the binding of nuclear proteins from B16 cells to the "M-box" region of the mouse tyrosinase promoter. Short-term PDBu treatment of B16 cells transfected with a mouse tyrosinase promoter-luciferase construct resulted in increased reporter gene activity, while long-term PDBu treatment inhibited reporter gene activity. Using an oligonucleotide containing the M-box and its flanking residues in electrophoretic mobility shift assays, we found a density-dependent change in the pattern of DNA-protein complexes. One complex was found to be negatively regulated by long-term PDBu treatment. Competition experiments with various mutated oligonucleotides demonstrated that both the M-box and flanking residues are important for nuclear protein binding. The complex whose formation was inhibited by long-term PDBu treatment was shown to contain the basic helix-loop helix leucine zipper protein microphthalmia-associated transcription factor (MITF). These results suggest that chronic PDBu treatment might inhibit tyrosinase expression (and subsequent melanogenesis) by affecting the amount or function of MITF. PMID- 9417871 TI - Nuclear Rad51 foci induced by DNA damage are distinct from Rad51 foci associated with B cell activation and recombination. AB - Lipopolysaccharide (LPS) is a B cell mitogen which can stimulate murine primary B cells to proliferate and carry out immunoglobulin heavy chain class switch recombination. LPS can also function as an endotoxin, which may cause DNA damage and apoptosis in certain types of cells. We have previously reported that LPS activated primary murine B cells contain nuclear foci that stain brightly with anti-Rad51 antibodies (Li et al. (1996) Proc. Natl. Acad. Sci. USA 93, 10222 10227). We have now analyzed Rad51 nuclear foci induced in both primary and immortalized B cells by treatment with the DNA damaging agent, methyl methanesulfonate (MMS). We have found that, in LPS-cultured primary B cells, MMS treatment increases the fraction of cells containing Rad51 foci and induces formation of a very high number of foci per cell. The foci induced by MMS treatment are small, punctate, and numerous; in contrast, the foci induced by LPS activation are large, brightly staining, and relatively few in number. In LPS cultured primary B cells, Rad51 relocalizes during the cell cycle, and large, brightly staining nuclear foci are present in only restricted stages of the cell cycle. Rad51 foci similar to those present in LPS-activated primary B cells are also observed in immortalized B cells lines cultured in the absence of LPS. These foci are unaltered in number or appearance by culture with LPS, but treatment of immortalized B cell lines with MMS induces foci which are small and punctate in staining, like those induced by MMS in primary B cells. These data show that distinctive Rad51 foci are induced by DNA damaging agents and cell activation and that the response to DNA damage may involve pathways distinct from those associated with B cell activation and switch recombination. PMID- 9417872 TI - Bcl-2 suppresses apoptosis resulting from disruption of the NF-kappa B survival pathway. AB - A role has been delineated for both bcl-2 and NF-kappa B in mediating an adaptive survival response to the TNF-alpha signaling pathway for apoptosis. Additionally, we and others have demonstrated a role for bcl-2 upregulation during progression of prostate cancer and acquisition of androgen-independent growth (T. J. McDonnell et al., 1992, Cancer Res. 52, 6940-6944). Therefore, the relationship between bcl-2 and NF-kappa B in regulating TNF-alpha-induced apoptosis was investigated in prostate carcinoma cells. Enforced overexpression of bcl-2 protein in prostatic carcinoma cells impaired TNF-alpha-mediated cytotoxicity. Expression of bcl-2 did not impose a block to, or potentiate, TNF-alpha signaling of I kappa B alpha degradation, nuclear import of the RelA p65, or NF-kappa B dependent transactivation. Expression of two dominant-negative I kappa B alpha mutant proteins significantly enhanced TNF-alpha-induced apoptosis in control cells but not in cells expressing high levels of bcl-2 protein. Similarly, PDTC, a strong antioxidant that interferes with activation of NF-kappa B in these prostate carcinoma cells, also potentiated TNF-alpha-stimulated apoptosis signaling through a bcl-2-regulated mechanism. These findings indicate that modulation of NF-kappa B survival signaling may be used to clinical advantage in the treatment of prostate cancer patients. The efficacy of strategies proposed to enhance TNF-alpha-mediated cytotoxicity by inhibiting NF-kappa B will likely be influenced by context-dependent variables such as bcl-2 expression. PMID- 9417873 TI - A GPIb alpha-related protein is expressed by fresh human breast carcinoma tissue and is regulated by a PKC-sensitive mechanism. AB - Fresh frozen breast carcinoma tissues were examined for the presence of GPIb alpha by immunohistochemistry. GPIb alpha was detected in six of seven primary invasive intraductal breast carcinoma tissues whereas staining was negative in seven of seven nonmalignant breast specimens. When biotin-labeled, triton-lysed, phorbol-12-myristate 13-acetate (PMA)-incubated breast carcinoma MCF-7 cells were immunoprecipitated with a MoAb directed against the platelet GPIb/IX complex, expression of GPIb was significantly enhanced in comparison to control preparations. Furthermore, incubation of MCF-7 cells for 84 h with 16 nmol/L PMA, but not with its biologically inactive derivative MePMA, induced a three- to fourfold increase in the surface expression of both GPIb alpha and GPIb/IX by flow cytometry. This PMA-enhanced GPIb alpha expression was almost completely abrogated when MCF-7 cells were first preincubated with the specific protein kinase C (PKC) inhibitor, H-7, prior to PMA treatment. Finally, PMA-incubated MCF 7 cells demonstrated a 63% (N = 6; P < 0.001) increase in tumor-induced platelet agglutination when added to platelets in comparison to control tumor cells. This enhancement could be abrogated by H-7. These findings confirm the expression of a protein with homology to platelet GPIb alpha expressed by fresh human breast carcinoma tissues, demonstrate that PMA enhances GPIb membrane expression by MCF 7 cells, and suggest that PKC plays a role in this process. PMID- 9417874 TI - Reduction of both RAR and RXR levels is required to maximally alter sensitivity of CA-OV3 ovarian tumor cells to growth suppression by all-trans-retinoic acid. AB - We wished to determine the effect of altering the levels or functional activity of retinoid receptors, in particular retinoic acid receptor-alpha (RAR-alpha) and retinoid X receptor-alpha (RXR-alpha) on the growth sensitivity of ovarian tumor cells to all-trans-retinoic acid (all-trans-RA). We found that CA-OV3 cells could be made resistant to all-trans-RA growth inhibition by overexpressing RAR beta(R269Q), an efficient dominant negative mutant which inhibits the function of all RAR subtypes. Antisense technology was then used to prepare stable transfectants of the retinoid-sensitive ovarian carcinoma cell line CA-OV3 in which expression of RAR-alpha, RXR-alpha, or both RAR-alpha and RXR-alpha was reduced. The effect of all-trans-RA on ovarian tumor cell growth was determined by MTT assay, autoradiographic analysis of DNA synthesis, and anchorage independent colony formation in soft agar. Our results show that cell lines expressing reduced levels of either RAR-alpha alone or RXR-alpha alone exhibited a small decrease in sensitivity to growth inhibition by all-trans-RA. However, maximum RA resistance was obtained in cell lines in which the levels of both RAR alpha and RXR-alpha were reduced. These results demonstrate the importance of both retinoid nuclear receptors and retinoid-X receptors in general, and RAR alpha and RXR-alpha in particular, as mediators of ovarian carcinoma cell growth inhibition by retinoids. PMID- 9417875 TI - Nuclear association of cyclin D1 in human fibroblasts: tight binding to nuclear structures and modulation by protein kinase inhibitors. AB - The association of cyclin D1 with nuclear structures was investigated in normal human fibroblasts by using hypotonic detergent extraction procedures, immunofluorescence quantitation with flow cytometry, and Western blot analysis. About 20% of the total cellular levels of cyclin D1 was found to be tightly bound to nuclear structures, being the complex formation resistant to DNase I treatment and to high salt extraction. Maximal levels of the insoluble form of the protein were found in the middle to late G1 phase of the cell cycle. Cell fractionation and immunoprecipitation techniques after in vivo 32P-labeling showed that both soluble and nuclear-bound forms of cyclin D1 were phosphorylated. Both fractions were reactive to an anti-phosphotyrosine antibody, while only the latter was detectable with an anti-phosphoserine antibody. Treatment with the protein kinase inhibitor staurosporine, which induces a cell cycle arrest in early G1 phase, strongly reduced cyclin D1 phosphorylation. Concomitantly, the ratio of nuclear bound/total cyclin D1 levels was reduced by about 60%, compared with the control value. The protein kinase A specific inhibitor isoquinoline-sulfonamide (H-89) induced a similar reduction in the ratio, with no significant modification in the total amount of protein. In contrast, both calphostin C and bisindolylmaleimide, specific inhibitors of protein kinase C, consistently increased by 30-50% the ratio of nuclear-bound/total amount of the cyclin protein. These results suggest that, during the G1 phase, formation of an insoluble complex of cyclin D1 occurs at nuclear matrix structures and that this association is mediated by a protein kinase A-dependent pathway. PMID- 9417876 TI - Synthesis of collagen by bovine chondrocytes cultured in alginate; posttranslational modifications and cell-matrix interaction. AB - The extracellular matrix synthesized by articular chondrocytes cultured in alginate beads was investigated. Collagen levels increased sigmoidally with time and remained constant after 2 weeks of culture. The presence of cartilage specific type II collagen was confirmed immunohistochemically. Predominantly type II collagen was present in the alginate bead, as reflected by the unique extent of lysyl hydroxylation, glycosylation, and pyridinoline crosslink formation measured. Collagen crosslinks, predominantly hydroxylysylpyridinoline (> 93%), were observed after 7 to 11 days of culture and their formation was effectively blocked by beta-aminopropionitrile (BAPN). Unexpectedly, BAPN treatment resulted in a 100% increase of collagen levels, without influencing cell proliferation and proteoglycan levels. In control cultures 90% of the synthesized collagen was retained in the cell-associated matrix, while in BAPN-treated cultures half of the collagen was found in the interterritorial matrix compartment further removed from the cells. This suggests that impaired crosslinking of collagen interferes with pericellular collagen deposition, causing upregulation of collagen synthesis by impaired cell-matrix interactions. Integrins are likely to be involved in this feedback inhibition by extracellular collagen since the cyclic RGD-containing peptide CGRGDSPC downregulated collagen synthesis by 37%. PMID- 9417877 TI - Division of polar bodies induced by their enlargement in the starfish Asterina pectinifera. AB - The first polar body (FPB), which is formed at the first meiotic division during oogenesis, does not generally divide. We made a hypothesis that the amount of cytoplasm was not sufficient for the FPB to perform cell division, in spite of the same amount of genomes and centrosome as those of the secondary oocyte. To examine this hypothesis, hexylene glycol (HG) at a low concentration was applied to oocytes of the starfish Asterina pectinifera during the first meiotic division. Hence, FPBs were enlarged in their diameters, some of them divided once, and the division rate increased in proportion as their diameter extended. We further hypothesized that the difference between the second polar body (SPB) and the egg would be only the amount of cytoplasm and that if SPBs were enlarged, they would become eggs. When the secondary oocytes were treated with HG, large SPBs were obtained. Some of them, however, divided, and resultant daughter cells divided moreover, whereas eggs would not cleave unless they were fertilized. We discuss here the reason why the centrosome distributed during maturation division began to function in enlarged PBs. PMID- 9417878 TI - Developmental regulation of Hsp32, a small heat shock protein in Dictyostelium discoideum. AB - We have examined the expression and heat inducibility of Hsp32, a novel small heat shock protein in Dictyostelium discoideum. Both Hsp32 and its mRNA are abundant in amoebae growing at physiological temperatures. Levels of Hsp32 remain high during the initial phases of development, including the formation of tipped mounds. After that stage, Hsp32 levels decrease, reaching barely detectable levels in culminating cells. In contrast, most of the hsp32 mRNA is rapidly degraded within the first few hours of starvation-induced development. Cells retain a new low steady state level of the mRNA throughout the rest of the developmental cycle. However, when cells undergo dedifferentiation, they reaccumulate high levels of hsp32 mRNA just prior to cell division. The heat inducibility of Hsp32 and its mRNA is maximal in growing cells and decreases as cells progress in their developmental program. The data suggest that Hsp32 is associated with a growth and/or survival function that is gradually eliminated during development. PMID- 9417879 TI - Differences in the structural characteristics of adult guinea pig and rat cardiomyocytes during their adaptation and maintenance in long-term cultures: confocal microscopy study. AB - In this study, we used laser confocal scanning microscopy and immunofluorescent markers to describe the establishment of long-term (1-5 week) cultures of adult guinea pig cardiomyocytes and adult rat cardiomyocytes. Providing that the preparation of freshly isolated guinea pig cardiomyocytes consists mostly (> 80%) of rod-shaped, Ca(2+)-tolerant, and quiescent cells and these are plated under optimal conditions and density (10(5) cells/cm2), these myocytes have the following characteristics: (1) they remain elongated with regular ultrastructural characteristics and quiescent for 1 week; (2) within 10-14 days, they reestablish intercellular contacts and resume contractile activity, which becomes synchronous all through the confluent layer; (3) their myofibrillar striations remain regular all through the adaptation to culture conditions without any sign of dedifferentiation or redifferentiation; (4) they form adherence junctions (as indicated by their immunoreactivity to an antibody against N-cadherin) over the entire cellular surface; (5) they appear to retain their ability to express atrial natriuretic peptide (ANP), as indicated by immunoreactivity to anti-ANP antibody; (6) this activity seems to be directly related to the surface area of the myocytes in contact with the substrate. In contrast, rat cardiomyocytes cultured under very similar and optimal conditions exhibit very different characteristics during their adaptation in long-term cultures: (1) although 85 90% of freshly isolated cells are also rod-shaped and Ca2+ tolerant they exhibit slow spontaneous contractions; (2) they round up during the first few days, and during the first week they dedifferentiate, losing their regular striated appearance; (3) they spread, becoming irregularly shaped, and, unlike the guinea pig cardiomyocytes, they do not form confluent layers, no matter what the plating density is; (4) they atrophy at a very early stage in the cultures, so that by the fourth week, they have lost most of their myofibrils; (5) the initial rounding up is largely eliminated by exposure for 24 h to 1 microM ryanodine or 20 mM butanedione monoxime, compounds that suppress the spontaneous contractions. In conclusion, our studies demonstrate that adult guinea pig cardiomyocytes adapt and survive in long-term (1-5 week) cultures much better than do adult rat cardiomyocytes, indicating that the long-term cultures of adult guinea pig ventricular myocytes provide a valuable experimental model which opens new possibilities for studying the cellular and molecular regulation of myocardial function under the acute or chronic influence of various intrinsic and/or extrinsic factors. PMID- 9417880 TI - Stimulation of oxygen consumption promotes mitochondrial calcium accumulation, a process associated with, and causally linked to, enhanced formation of tert butylhydroperoxide-induced DNA single-strand breaks. AB - The NADH-linked substrates pyruvate, L-glutamine, and beta-hydroxybutyrate, while enhancing the rate of oxygen consumption, also increased the formation of DNA single-strand breaks induced by tert-butylhydroperoxide in intact U937 cells. A cause-effect relationship between these two parameters was established by showing that: (a) rotenone, an inhibitor of complex I, abolished respiration and prevented the enhancement of the DNA-damaging response under all the above circumstances; (b) the membrane-impermeant, complex I-activating substrate L malate gave similar results in permeabilized cells; and (c) none of the NADH linked substrates affected the DNA-damaging response to tert-butylhydroperoxide in respiration-deficient cells. Stimulation of electron transport potentiated the DNA-cleaving ability of tert-butylhydroperoxide via a process involving enforced mitochondrial calcium accumulation in the absence of a discernible elevation in the cytosolic concentration of free Ca2+. Finally, mitochondrial calcium was found to promote the mitochondrial formation of DNA-damaging levels of hydrogen peroxide. In conclusion, the data herein presented define a previously unexpected role of respiratory substrates in the control of the deleterious effects of an organic hydroperoxide at the level of genomic DNA. The enhanced DNA cleavage mediated by NADH-linked substrates in response to tert-butylhydroperoxide would appear to depend on a sequence of events involving stimulation of electron transport, mitochondrial accumulation of Ca2+, and mitochondrial formation of DNA damaging levels of hydrogen peroxide via a Ca(2+)-dependent process. PMID- 9417881 TI - Bifunctional effects of a protein kinase inhibitor (H-7) on heat-induced p53 dependent WAF1 accumulation. AB - We have previously shown that heat shock induces p53-dependent WAF1 expression. To understand the role of protein kinases in the heat-induced p53-mediated signal transduction pathway, the effects of H-7, a serine/threonine kinase inhibitor, on WAF1 accumulation were investigated using two human glioblastoma cell lines differing in p53 status or their transfectants with various p53-expression vectors. Unexpectedly, H-7 alone induced p53-dependent WAF1 accumulation with a biphasic pattern depending on H-7 dose; i.e., low doses of H-7 induced the accumulation of both p53 and WAF1, whereas, a high dose of H-7 induced p53 but no WAF1 accumulation, suggesting that p53 accumulation and p53-dependent WAF1 expression are separable. Heat shock and H-7 induce p53-dependent WAF1 accumulation through different pathways as shown in A-172 cells stably expressing a temperature-sensitive mutant p53. However, our results show that these two pathways cross-talk with each other in the combined treatment of H-7 and heat shock studies. These findings indicate that inhibition of protein kinases can act as a novel stress to evoke the p53 pathway and that p53 activation by heat shock requires activation of yet unidentified protein kinases in vivo. The cross-talks between H-7 and heat shock in the p53 pathway provide the first evidence for the complex interactions between different stress signaling pathways in the modulation of p53 in vivo. PMID- 9417882 TI - Developmental regulation of Mac25/insulin-like growth factor-binding protein-7 expression in skeletal myogenesis. AB - Mac25 is a newly discovered member of the insulin-like growth factor-binding protein (IGFBP) family, recently assigned the name IGFBP-7, Mac25/IGFBP-7 is hypothesized to have growth-suppressing activity, since mac25/IGFBP-7 mRNA is down-regulated in several tumor cell lines and is highly expressed in senescent mammary epithelial cells. In this study, mac25/ IGFBP-7 mRNA expression was characterized in the C2 skeletal myogenic cell line, which undergoes a transition from actively dividing, undifferentiated myoblasts to nondividing, differentiated myotubes. Mac25/ IGFBP-7 mRNA levels were 2.5-fold higher in dividing C2 myoblasts than in nondividing myotubes. The inverse correlation between mac25/IGFBP-7 expression and myogenic differentiation was further examined by treating myogenic cultures with transforming growth factor-beta (TGF-beta) or insulin-like growth factor-I (IGF-I). TGF-beta inhibited myogenic differentiation by 98% and stimulated mac25/IGFBP-7 mRNA expression 2-fold. IGF-I stimulated differentiation by 50% and inhibited mac25/IGFBP-7 expression 2- to 3-fold. These findings indicate that, in contrast to other cell systems examined so far, expression of this new member of the IGFBP family is not always correlated with a nonproliferative state. PMID- 9417883 TI - Mode of the autocrine/paracrine mechanism of growth hormone action. AB - GH is synthesized at multiple extrapituitary sites suggestive of an autocrine/paracrine mechanism of action. We have investigated a possible autocrine/paracrine mechanism of GH action, compared the cellular response to exogenous versus endogenously produced GH, and determined the nature of the interaction between external stimuli and endogenously produced GH. BRL cells expressing the GH receptor were transiently transfected with expression plasmids containing either the hGH or the bGH gene and the response of the cell was measured by CAT reporter plasmids requiring either STATs 1 and 3 or STAT5 for their response. Transient transfection of the hGH gene resulted in hGH accumulation in the cell and secretion into the media. The functional response through STATs 1 and 3 and STAT5 obtained with endogenously produced hGH was comparable or greater in magnitude to that obtained with the maximal stimulatory dose of exogenous hGH. Similar results were obtained with an expression plasmid containing the bGH gene. Endogenously produced hGH interacted in an additive manner when combined with submaximal doses of both exogenous hGH and serum. Such results were also observed in a more physiologically relevant mammary carcinoma cell line (MCF-7). The nonreceptor-dimerizing hGH antagonist, hGH-G120R, used in cells expressing the homologous receptor extracellular domain was able to only partially inhibit the response of the cell to endogenously produced hGH, in contrast to full inhibition of exogenous hGH. We therefore conclude that GH can function in an autocrine/paracrine manner, additive in effect to external stimuli. PMID- 9417884 TI - Cystine starvation induces reversible large-body formation from nuclear bodies in T24 cells. AB - Nuclear bodies (PML nuclear bodies; also referred to as PODs or ND10) are small intranuclear structures which contain PML as an essential constituent and from several to 20 or more of them are present per nucleus. When starved of amino acids, nuclear bodies reversibly form one or more doughnut-shaped large bodies in T24 cells. Here, I present evidence that cystine is essential for the maintenance of normal-sized nuclear bodies: Cystine deficiency induces the formation of large bodies, and the addition of cystine or cysteine induces the reappearance of many normal-sized nuclear bodies. Both PML and Sp100 exhibit the same behavior as that of the nuclear body antigen(s) of AP435 MAb with regard to these changes, L buthionine-[S,R]-sulfoximine, an inhibitor of glutathione synthesis, also induced the formation of large bodies that could be reversed to nuclear bodies by both beta-mercaptoethanol and dithiothreitol as well as cystine. Large bodies have the characteristic electron-dense, spherical, and concentric or reticulated features of nuclear bodies as observed by electron microscopy. The results indicate that environmental cystine controls reversible change between nuclear bodies and large bodies, which are not mere aggregates of nuclear bodies, but rather reconstituted nuclear bodies. PMID- 9417885 TI - Interferon-gamma inhibits insulin release and induces cell death in the pancreatic beta-cell line INS-1 independently of nitric oxide production. AB - Interferon-gamma is among the cytokines which have been implicated as effector molecules of beta-cell destruction in autoimmune diabetes. Its mechanism of action is, however, largely unknown. In the present study rat pancreatic beta cells, INS-1, were incubated with rat interferon-gamma (rIRN-gamma) for 24 h. rIFN-gamma at 1-1000 U/ml caused a dose-dependent inhibition of insulin release and cell metabolism with maximal inhibition being observed at 100 U/ml (insulin release: 51.2%, cell metabolism: 43.3% of control, respectively). In addition, 100 U/ml rIFN-gamma induced a 4- and 8.3-fold increase in apoptotic cell death after 24 and 48 h of incubation, respectively. These effects were not mediated by nitric oxide (NO), since IFN-gamma failed to induce nitric oxide synthase and NO production. Similarly, beta-cell dysfunction and death were not prevented by coincubation of the INS-1 cells with the poly(ADP-ribose) polymerase inhibitors benzamide, 3-aminobenzamide, and 4-aminobenzamide, the oxygen free radical scavenger Trolox, and the antioxidant N-acetylcysteine, indicating that NO, poly(ADP-ribose) polymerase, and oxygen free radicals are not involved in IFN gamma induced beta-cell dysfunction and death. PMID- 9417886 TI - Hyaluronan oligosaccharides induce tube formation of a brain endothelial cell line in vitro. AB - In remodeling tissues the formation of new blood vessels is an essential process which is regulated by different factors. During such processes an increase in hyaluronan synthesis and turnover is often seen and recent observations have suggested that hyaluronan can both promote and inhibit neovascularization depending on its molecular mass. In this work we show that a brain capillary endothelial cell line forms tubes in a collagen gel after stimulation by hyaluronan oligosaccharides. Ultrastructural examination of the tubes by transmission electron microscopy revealed that the cord-like outgrowths consisted of 2-10 tightly packed cells containing a continuous lumen. The tube formation in response to hyaluronan oligosaccharide was not mediated by activation of receptors for fibroblast growth factor. The endothelial cell line, which does not synthesize hyaluronan, exhibited specific hyaluronan-binding sites, with about 3000 hyaluronan molecules (M(r) 3.85 x 10(6)) bound per cell at saturation and Kd was 0.05 x 10(-9) M. Furthermore, the cell line was stained with mAb IVd4 that recognizes a novel class of hyaluronan-binding proteins and mAb IM7 which recognizes CD44 molecules. PMID- 9417887 TI - Anti-IgM-induced growth inhibition and apoptosis are independent of ornithine decarboxylase in Ramos cells. AB - Ornithine decarboxylase (ODC) is a key enzyme involved in polyamine production and is thought to regulate growth and apoptosis in multiple cell systems. A potential link between ODC and growth may involve the action of an oncogene c-myc which is thought to transcriptionally regulate ODC. We have examined the involvement of ODC in anti-IgM-induced growth inhibition and apoptosis in Burkitt's lymphoma cells. Inhibitors of ODC such as difluoromethylornithine (DFMO) completely blocked ODC activity, resulting in growth inhibition but not apoptosis. Addition of putrescine, the product of ODC enzymatic action, to Ramos cells had only a minor effect on growth, did not cause apoptosis, did not augment or block anti-IgM-mediated growth inhibition and apoptosis, but did reverse DFMO mediated growth inhibition. Anti-IgM treatment of Ramos cells, which markedly decreased c-myc mRNA and protein, caused a paradoxical increase in ODC mRNA level as well as ODC enzymatic activity and increased cellular levels of putrescine. DFMO and putrescine did not alter c-myc mRNA levels directly, nor did they have any affects on anti-IgM-mediated down-regulation of c-myc mRNA. TNF-alpha, which inhibited anti-IgM-mediated apoptosis, did not inhibit either anti-IgM or DFMO mediated inhibition of growth. These agents were without effect on ODC activity itself or on the anti-IgM-mediated increase in ODC activity. From these studies we conclude that ODC inhibition affects growth but is unrelated to the induction of apoptosis. Both anti-IgM-mediated inhibition of growth and induction of apoptosis are independent of ODC. Thus two distinct pathways for growth regulation are present: one in which ODC and polyamines are important and the other cell surface receptor-mediated (sIg) which is independent of ODC and polyamines. PMID- 9417888 TI - The effect of lysosomotropic amines on beige mouse cells. AB - The beige mutant in the mouse is characterized by enlarged lysosomes in many cell types due to increased fluidity of cellular membranes and organelle fusion. In this study, mutant fibroblasts derived from C57BL/ 6J;bgJ/bgJ animals were compared with control fibroblasts (C57BL/6J;+/+) for vacuolation when treated with lysosomotropic weak bases including ammonium chloride, trimethylamine, and methylamine. These amines produce vacuolation by their accumulation in cellular acidic compartments, which causes osmotic swelling and fusion of organelles due to the increase in pH. Beige cells exhibited greater vacuolation than control cells for all treatments, which was indicative of the mutant's effect on organelle fusion and membrane fluidity. Trimethylamine caused the most pronounced difference in vacuolation between mutant and control cells. This method allows for a simple morphological distinction between beige and control cells that also utilizes a physiological difference in the expression of the beige gene. PMID- 9417889 TI - Phylogeny of the avian family Ciconiidae (storks) based on cytochrome b sequences and DNA-DNA hybridization distances. AB - This study is a phylogenetic analysis of the avian family Ciconiidae, the storks, based on two molecular data sets: 1065 base pairs of sequence from the mitochondrial cytochrome b gene and a complete matrix of single-copy nuclear DNA DNA hybridization distances. Sixteen of the nineteen stork species were included in the cytochrome b data matrix, and fifteen in the DNA-DNA hybridization matrix. Both matrices included outgroups from the families Cathartidae (New World vultures) and Threskiornithidae (ibises, spoonbills). Optimal trees based on the two data sets were congruent in those nodes with strong bootstrap support. In the best-fit tree based on DNA-DNA hybridization distances, nodes defining relationships among very recently diverged species had low bootstrap support, while nodes defining more distant relationships had strong bootstrap support. In the optimal trees based on the sequence data, nodes defining relationships among recently diverged species had strong bootstrap support, while nodes defining basal relationships in the family had weak support and were incongruent among analyses. A combinable-component consensus of the best-fit DNA-DNA hybridization tree and a consensus tree based on different analyses of the cytochrome b sequences provide the best estimate of relationships among stork species based on the two data sets. PMID- 9417890 TI - Phylogenetic analysis of the Arenaviridae: patterns of virus evolution and evidence for cospeciation between arenaviruses and their rodent hosts. AB - Viruses of the Arenaviridae cause hemorrhagic fevers and neurologic disease in humans. Historically, the arenaviruses have been divided into two complexes (LASV LCMV, Tacaribe) through the use of antigenic typing. The phylogeny of the Arenaviridae as a whole has not been estimated previously due to a lack of sequence data for all members of the family. In this study, nucleocapsid protein gene sequence data were obtained for all currently known arenaviruses and used to estimate, for the first time, a phylogeny of the entire virus family. The LCMV LASV complex arenaviruses are monophyletic and comprise three distinct lineages. The Tacaribe complex viruses also are monophyletic and occupy three distinct lineages. Comparisons of arenavirus phylogeny with rodent host phylogeny and taxonomic relationships provide several examples in which virus-host cospeciation is potentially occurring. The pathogenic arenaviruses do not appear to be monophyletic, suggesting that the pathogenic phenotype has arisen in multiple independent events during virus evolution. PMID- 9417891 TI - The potential of Betv1 homologues, a nuclear multigene family, as phylogenetic markers in flowering plants. AB - Betv1 homologues are a ubiquitous group of genes in flowering plants encoding a class of highly conserved defense-related proteins and containing open reading frames from 465 to 480 bp. Betv1-like genes consist of two exons interrupted by an intron of 76-359 bp, with the intron position highly conserved. The pairwise p distance ranged from 0 to 0.583 among flowering plants. Within plant families, the ranges of the p distance were 0-0.403, 0-0.253, and 0.011-0.369, for Apiaceae, Betulaceae, and Fabaceae, respectively. The most striking feature of the betv1 gene phylogeny was that the multiple sequences from each plant family formed a monophyletic group and sequences from each species were generally more similar to each other than those from other species. The almost exclusive paralogous relationships of genes from the same species suggested that the genes of the multigene family underwent strong concerted evolution. Phylogenies of Betulaceae and Fabaceae inferred from betv1 gene trees were generally congruent with those based on morphology and other molecules. Betv1 homologues constitute potential phylogenetic markers at the intrafamilial level or among closely related families in flowering plants. PMID- 9417892 TI - Molecular systematics of tanagers (Thraupinae): evolution and biogeography of a diverse radiation of neotropical birds. AB - The tanagers (Passeriformes: Emberizidae: Thraupinae) are a diverse group of mostly Neotropical birds with a wide range of feeding morphologies, behaviors, plumage patterns and colors, and habitat preferences. Phylogenetic relationships of genera in this lineage were investigated using cytochrome b sequence data. This study indicates that the genera Euphonia and Chlorophonia (traditionally considered part of Thraupinae) do not form a monophyletic group with the other tanagers. Within the rest of Thraupinae, several monophyletic groups are identified that agree with traditional sequential taxonomies. Other monophyletic groups provide novel interpretations of biogeographic patterns and morphological evolution within tanagers. In several lineages, plumage patterns and colors persist despite dramatic changes in bill morphology. Phylogenetic structure and estimated timings of divergence events indicate that tanagers probably originated on Caribbean islands and later diversified throughout Central and South America during the mid-Tertiary. PMID- 9417893 TI - Inferring species trees from gene trees: a phylogenetic analysis of the Elapidae (Serpentes) based on the amino acid sequences of venom proteins. AB - Toward the goal of recovering the phylogenetic relationships among elapid snakes, we separately found the shortest trees from the amino acid sequences for the venom proteins phospholipase A2 and the short neurotoxin, collectively representing 32 species in 16 genera. We then applied a method we term gene tree parsimony for inferring species trees from gene trees that works by finding the species tree which minimizes the number of deep coalescences or gene duplications plus unsampled sequences necessary to fit each gene tree to the species tree. This procedure, which is both logical and generally applicable, avoids many of the problems of previous approaches for inferring species trees from gene trees. The results support a division of the elapids examined into sister groups of the Australian and marine (laticaudines and hydrophiines) species, and the African and Asian species. Within the former clade, the sea snakes are shown to be diphyletic, with the laticaudines and hydrophiines having separate origins. This finding is corroborated by previous studies, which provide support for the usefulness of gene tree parsimony. PMID- 9417894 TI - Cytochrome B sequences suggest convergent evolution of the Asian takin and Arctic muskox. AB - Relationships of the takin (Budorcas taxicolor) and muskox (Ovibos moschatus) have been speculated upon for many years. Morphological and behavioral similarities between these species have led to suggestions that they are closely related. To test the hypothesis that characteristics shared by the takin and muskox stem from a recent common ancestor, we compared sequences of their mitochondrial cytochrome b genes with those of three other species of Caprinae. We present data that may support rejection of the hypothesis of recent common ancestry and suggest that similarities in behavior and morphology in these two species might be attributed to convergent evolution rather than shared phylogeny. PMID- 9417895 TI - Escaping from the Felsenstein zone by detecting long branches in phylogenetic data. AB - Long branches in a true phylogeny tend to disrupt hierarchical character covariation (phylogenetic signal) in the distribution of traits among organisms. The distortion of hierarchical structure in character-state matrices can lead to errors in the estimation of phylogenetic relationships and inconsistency of methods of phylogenetic inference. Examination of trees distorted by long-branch attraction will not reveal the identities of problematic taxa, in part because the distortion can mask long branches by reducing inferred branch lengths and through errors in branching order. Here we present a simple method for the detection of taxa whose placement in evolutionary trees is made difficult by the effects of long-branch attraction. The method is an extension of a tree independent conceptual framework of phylogenetic data exploration (RASA). Taxa that are likely to attract are revealed because long branches leave distinct footprints in the distribution of character states among taxa, and these traces can be directly observed in the error structure of the RASA regression. Problematic taxa are identified using a new diagnostic plot called the taxon variance plot, in which the apparent cladistic and phenetic variances contributed by individual taxa are compared. The procedure for identifying long edges employs algorithms solved in polynomial time and can be applied to morphological, molecular, and mixed characters. The efficacy of the method is demonstrated using simulated evolution and empirical evidence of long branches in a set of recently published sequences. We show that the accuracy of evolutionary trees can be improved by detecting and combating the potentially misleading influences of long branch taxa. PMID- 9417896 TI - Molecular evolution and phylogenetic utility of the chloroplast rpl16 intron in Chusquea and the Bambusoideae (Poaceae). AB - Phylogenetic relationships within Chusquea, a diverse genus of neotropical woody bamboos, and among selected members of the Bambusoideae were explored using rpl16 intron sequence data from the chloroplast genome. Mechanisms of mutation, including slipped-strand mispairing, secondary structure, minute inversions, and base substitutions, were examined within the rpl16 intron, and their effects on sequence alignment and phylogenetic analysis were investigated. Thirty-five bamboo sequences were generated and two separate matrices were analyzed using maximum parsimony. In the first, 23 sequences from Chusquea, 1 of Neurolepis, and 3 outgroups were included. Neurolepis was supported as sister to Chusquea, Chusquea was strongly supported as a monophyletic lineage, and three species of Chusquea subg. Rettbergia were resolved as the most basal clade within the genus. In the second analysis, 15 sequences, 14 from across the subfamily and 1 outgroup, were included. A Bambusoideae clade was recovered with the Olyreae/Parianeae (herbaceous bamboos) and the Bambuseae (woody bamboos) each supported as monophyletic. Two clades corresponding to temperate and tropical woody bamboos were derived within the Bambuseae and the tropical taxa were further split into New World and Old World clades. The rpl16 intron in bamboos was found to be susceptible to frequent length mutations of multiple origins, nonindependent character evolution, and regions of high mutability, all of which created difficulties in alignment and phylogenetic analysis; nonetheless the rpl16 intron is phylogenetically informative at the inter- and intrageneric levels in bamboos. PMID- 9417897 TI - General time-reversible distances with unequal rates across sites: mixing gamma and inverse Gaussian distributions with invariant sites. AB - A series of new results useful to the study of DNA sequences using Markov models of substitution are presented with proofs. General time-reversible distances can be extended to accommodate any fixed distribution of rates across sites by replacing the logarithmic function of a matrix with the inverse of a moment generating function. Estimators are presented assuming a gamma distribution, the inverse Gaussian distribution, or a mixture of either of these with invariant sites. Also considered are the different ways invariant sites may be removed and how these differences may affect estimated distances. Through collaboration, we implemented these distances into PAUP in 1994. The variance of these new distances is approximated via the delta method. It is also shown how to predict the divergence expected for a pair of sequences given a rate matrix and a distribution of rates across sites, allowing iterated ML estimates of distances under any reversible model. A simple test of whether a rate matrix is time reversible is also presented. These new methods are used to estimate the divergence time of humans and chimps from mtDNA sequence data. These analyses support suggestions that the human lineage has an enhanced transition rate relative to other hominoids. These studies also show that transversion distances differ substantially from the overall distances which are dominated by transitions. Transversions alone apparently suggest a very recent divergence time for humans versus chimps and/or a very old (> 16 myr) divergence time for humans versus orangutans. This work illustrates graphically ways to interpret the reliability of distance-based transformations, using the corrected transition to transversion ratio returned for pairs of sequences which are successively more diverged. PMID- 9417898 TI - Molecular evolution of the cottoid fish endemic to Lake Baikal deduced from nuclear DNA evidence. AB - Lake Baikal in Eastern Siberia contains a remarkable flock of 29 species of teleost fishes of the suborder Cottoidei (sculpins, bullheads) that are endemic to the lake and its associated rivers and occupy all depth habitats down to over 1500 m. The species are divided into three families, the Cottidae with 7 species, the Abyssocottidae with 20 species, and the Comephoridae with 2 species. Nucleotide sequences of the rod opsin gene from 12 of these species, plus a non Baikal marine species, have been used to examine the evolutionary relations and the divergence time of the flock. Phylogenetic trees, generated by neighbor joining and maximum parsimony, indicate that the unique Comephoridae family with its viviparity and unusual appearance is closely related to the Cottidae and Abyssocottidae, whereas the genus Cottocomephorus, at present placed in the Cottidae, was the first to diverge from the ancestral species and forms a separate lineage. The major adaptation to deep water would appear to be of relatively recent origin, and there is evidence that the ancestral species occupied a shallow-water-marine or brackish habitat. Estimates of antiquity obtained from synonymous substitutions place the origin of the species flock at around 4.9 million years ago. PMID- 9417899 TI - Molecular phylogeny of rodents, with special emphasis on murids: evidence from nuclear gene LCAT. AB - Phylogenetic relationships among 19 extant species of rodents, with special emphasis on rats, mice, and allied Muroidea, were studied using sequences of the nuclear protein-coding gene LCAT (lecithin:cholesterol acyltransferase), an enzyme of cholesterol metabolism. Analysis of 705 base pairs from the exonic regions of LCAT confirmed known groupings in and around Muroidea. Strong support was found for the families Sciuridae (squirrel and marmot) and Gliridae (dormice) and for suprafamilial taxa Muroidea and Caviomorpha (guinea pig and allies). Within Muroidea, the first branching leads to the fossorial mole rats Spalacinae and bamboo rats Rhizomyinae. The other Muroidea appear as a polytomy from which are issued Gerbillinae (gerbils), Murinae (rats and mice), Sigmodontinae (New World cricetids), Cricetinae (hamsters), and Arvicolinae (voles). Evidence from LCAT sequences agrees with that from a number of previous molecular and morphological studies, both concerning branching orders inside Muroidea and the bush-like radiation of rodent suprafamilial taxa (caviomorphs, sciurids, glirids, muroids), thus suggesting that this nuclear gene is an appropriate candidate for addressing questions of rodents relationships. PMID- 9417900 TI - Phylogenetic relationships within genus Leuciscus (Pisces, Cyprinidae) in Portuguese fresh waters, based on mitochondrial DNA cytochrome b sequences. AB - To investigate phylogenetic relationships among Leuciscus species occurring in Portuguese inland waters, the cytochrome b gene was sequenced from representatives of the main rivers. This study supports the recognition of the species level for L. pyrenaicus, including populations from the southern Portuguese drainages (Tejo, Sado, and Guadiana drainages), and for L. carolitertii, including populations from the northern Portuguese drainages. The existence of two new species occurring in the extreme southwestern drainages of Mira and Arade is also suggested. The present results support the monophyly of the Mira and the Arade populations, as well as an early divergence of these two lineages. The present-day distribution of Leuciscus species is seen as a consequence of Pliocene and Pleistocene events, such as river disjunctions and posterior confluence in epicontinental seas and river captures. A mixture of haplotypes was observed in the Mondego and the Tejo drainages, which could be a consequence of ancient river captures, with a possible mitochondrial DNA introgression in the Tejo drainage and a recent introduction by man in the Mondego drainage. The pattern of differentiation among mtDNA haplotypes and their geographic distribution is discussed in terms of evolutionary aspects. PMID- 9417901 TI - Re: Paralinear distance (LogDet) is a generalized correlation. PMID- 9417902 TI - Complementary uses of molecules and morphology: a reply to Lee. PMID- 9417903 TI - Vertebrate genome evolution--the decade ahead. PMID- 9417904 TI - Mapping and characterization of novel (CAG)n repeat cDNAs from adult human brain derived by the oligo capture method. AB - The expansion of a (CAG)n trinucleotide repeat has been associated with at least eight neurological disorders in which the repeats code for polyglutamine in the protein. To identify additional genes that possess (CAG)n repeats, single stranded cDNA clones derived from adult human brain were screened using biotinylated oligonucleotide (CAG)8, and the hybridizing complexes were isolated with strepavidin-coated paramagnetic beads. A total of 119 cDNA clones were isolated and initially characterized by end sequencing. BLAST homology searches were used to reduce redundancies with overlapping clones and to eliminate those that show sequence identity with previously published cDNAs with triplet repeats. Only cDNA clones with more than five CAG repeats were pursued for analysis. A total of 19 novel cDNAs were further characterized by determining chromosomal assignments using the Stanford G3 and Genebridge radiation-reduced hybrid mapping panels. Transcript sizes and tissue expression patterns were determined by Northern blot analysis. Two of 19 clones showed specific or high expression in brain. These cDNAs are ideal candidate genes for other neurodegenerative disorders, such as spinocerebellar ataxia types 5 and 7, and may also be implicated in psychiatric diseases such as bipolar affected disorder and schizophrenia. PMID- 9417905 TI - Construction of a 780-kb PAC, BAC, and cosmid contig encompassing the minimal critical deletion involved in B cell chronic lymphocytic leukemia at 13q14.3. AB - A putative tumor suppressor gene involved in B cell chronic lymphocytic leukemia (B-CLL) was mapped to human chromosome 13q14.3 close to the genetic markers D13S25 and D13S319. We constructed a 780-kb-long contig composed of cosmids, bacterial artificial chromosomes, and bacteriophage P1-derived artificial chromosomes that provides essential information and tools for the positional cloning of this gene. The conting contains both flanking markers as well as several additional genetic markers, three ESTs, and one potential CpG island. In addition, using one B-CLL patient, we characterized a small internal deleted region of 550 kb. Comparing this deletion with other recently published deletions narrows the minimally deleted area to less than 100 kb in our physical map. This deletion core region should contain all or part of the disrupted in B cell malignancies tumor suppressor gene. PMID- 9417907 TI - Analysis of protein domain families in Caenorhabditis elegans. AB - The Caenorhabditis elegans genome sequencing project has completed over half of this nematode's 100-Mb genome. Proteins predicted in the finished sequence have been compiled and released in the data-base Wormpep. Presented here is a comprehensive analysis of protein domain families in Wormpep 11, which comprises 7299 proteins. The relative abundance of common protein domain families was counted by comparing all Wormpep proteins to the Pfam collection of protein families, which is based on recognition by hidden Markov models. This analysis also identified a number of previously unannotated domains. To investigate new apparently nematode-specific protein families, Wormpep was clustered into domain families on the basis of sequence similarity using the Domainer program. The largest clusters that lacked clear homology to proteins outside Nematoda were analyzed in further detail, after which some could be assigned a putative function. We compared all proteins in Wormpep 11 to proteins in the human, Saccharomyces cerevisiae, and Haemophilus influenzae genomes. Among the results are the estimation that over two-thirds of the currently known human proteins are likely to have a homologue in the whole C. elegans genome and that a significant number of proteins are well conserved between C. elegans and H. influenzae, that are not found in S. cerevisiae. PMID- 9417906 TI - An ancient conserved gene expressed in the human inner ear: identification, expression analysis, and chromosomal mapping of human and mouse antiquitin (ATQ1). AB - We constructed and screened a human fetal cochlear cDNA library to identify genes involved in hearing and deafness. From this library we isolated a cDNA corresponding to the highly conserved ancient gene antiquitin (ATQ1). The plant homolog of ATQ1 is thought to be involved in regulating turgor pressure, a function that also would be essential for cells of the mammalian cochlea. Northern blots of 13 human fetal tissues show antiquitin to be highly expressed in cochlea, ovary, eye, heart, and kidney. Using RT-PCR of rat cochlear hair cell specific cDNA libraries, we detect antiquitin expression in outer hair cells, but not in inner or vestibular type 1 hair cells, suggesting that antiquitin is not expressed ubiquitously in the cochlea. Human ATQ1 was mapped to human chromosome region 5q31 using fluorescence in situ hybridization, and mouse ATQ1 was mapped to mouse chromosome 18 by single-strand conformation polymorphism mapping of interspecific backcross progeny DNAs. Four human antiquitin-like sequences, possibly pseudogenes, were also identified and mapped. PMID- 9417908 TI - Molecular cloning and characterization of LOH11CR2A, a new gene within a refined minimal region of LOH at 11q23. AB - Deletions at chromosome 11q23 are frequent events in a variety of human neoplasms, including breast, lung, and ovarian carcinomas. Two common regions of loss of heterozygosity, shared between lung and breast carcinomas, have been previously identified at 11q23, suggesting that the same tumor susceptibility genes are altered in these two malignancies. One of these regions, refined in lung adenocarcinoma, is included between loci D11S1345 and D11S1328. Here, we describe the refinement of the same region in breast carcinomas and the characterization of a new gene found within this area. The gene, called LOH11CR2A, spans an area of approximately 40 kb and is transcribed at a low level in all the tissues in which it has been analyzed. The predicted amino acid primary sequence revealed no homology with other proteins that could help to elucidate a possible function for the LOH11CR2A protein. Here, we tested the hypothesis that LOH11CR2A could be a tumor suppressor gene. Analysis of human breast, lung, and ovarian carcinomas revealed the presence of several amino acid substitutions in the coding region of this gene. However, a role for these amino acid changes in the tumorigenic process could not established. PMID- 9417909 TI - The mouse homeobox gene, Gbx2: genomic organization and expression in pluripotent cells in vitro and in vivo. AB - The Gbx2 homeodomain is widely conserved in metazoans. We investigated the mouse Gbx2 locus by isolation and characterization of genomic clones and by physical localization to the genome. The Gbx2 gene contained a single intron that separated the proposed functional protein domains. This organization was conserved with human GBX2. Physical localization of Gbx2 to Chromosome 1C5-E1 indicated that the genomic relationship between the linked Gbx2 and En1 genes differs between mouse and human, making it unlikely to be of functional significance. We also extended the known expression pattern of Gbx2 beyond the gastrulation stage embryo and the developing CNS to pluripotent cells in vitro and in vivo. Gbx2 expression was demonstrated in undifferentiated embryonic stem cells but was downregulated in differentiated cell populations. In the embryo, Gbx2 expression was detected before primitive streak formation, in the inner cell mass of the preimplantation embryo. Gbx2 is therefore a candidate control gene for cell pluripotency and differentiation in the embryo. PMID- 9417910 TI - Cloning, sequencing, gene organization, and localization of the human ribosomal protein RPL23A gene. AB - The intron-containing gene for human ribosomal protein RPL23A has been cloned, sequenced, and localized. The gene is approximately 4.0 kb in length and contains five exons and four introns. All splice sites exactly match the AG/GT consensus rule. The transcript is about 0.6 kb and is detected in all tissues examined. In adult tissues, the RPL23A transcript is dramatically more abundant in pancreas, skeletal muscle, and heart, while much less abundant in kidney, brain, placenta, lung, and liver. A full-length cDNA clone of 576 nt was identified, and the nucleotide sequence was found to match the exon sequence precisely. The open reading frame encodes a polypeptide of 156 amino acids, which is absolutely conserved with the rat RPL23A protein. In the 5' flanking region of the gene, a canonical TATA sequence and a defined CAAT box were found for the first time in a mammalian ribosomal protein gene. The intron-containing RPL23A gene was mapped to cytogenetic band 17q11 by fluorescence in situ hybridization. PMID- 9417911 TI - Genomic organization of the mouse reelin gene. AB - Reelin is the protein defective in reeler mice, an extensively studied model of brain development. The reelin gene (symbol Reln) codes for a protein of the extracellular matrix that contains eight successive repeats of 350 to 390 amino acids. In this work, we describe the genomic structure of the mouse reelin gene and the 5'-flanking genomic DNA sequences. The reelin gene is composed of 65 exons spread over approximately 450 kb of genomic DNA. We identified different reelin transcripts, formed by alternative splicing of a microexon as well as by use of two different polyadenylation sites. All splice sites conform to the GT-AG rule, except for the splice donor site of intron 30, which is GC instead of GT. A processed pseudogene is present in intron 42. Its nucleotide sequence is 86% identical to the sequence of the rat RDJ1 cDNA, which codes for a DnaJ-like protein of the Hsp40 family. Comparison of 8 intron positions in mouse and human reelin genes reveals a highly conserved genomic structure, suggesting a similar structure of the whole gene in both species. We identified two transcription start sites embedded within a CpG. The promoter region contains putative recognition sites for the transcription factors Sp1 and AP2 but lacks TATA and CAAT boxes. The presence of tandemly repeated regions in the Reelin protein suggests that gene duplication events occurred during evolution. By comparison of the amino acid sequences of the eight repeats and the positions of introns, we suggest a model for the evolution of the repeat coding portion of the reelin gene from a putative ancestral minigene. PMID- 9417912 TI - Identification of a novel gene, PSD, adjacent to NFKB2/lyt-10, which contains Sec7 and pleckstrin-homology domains. AB - We have identified a novel human gene on chromosome 10q24 located contiguously to the 3' end of the NFKB2/lyt-10 gene in a tail to tail arrangement. We describe here a cDNA of 4307 bp, isolated from an adult human brain cDNA library, which contains an open reading frame encoding a putative protein of 645 amino acids with a predicted molecular weight of 71 kDa. Database homology searches indicate that this is a novel gene coding for a putative protein containing two discrete domains with significant homology to the Sec7 and pleckstrin-homology (PH) domains, respectively. We named this gene PSD (plekstrin-Sec7 domains gene). Northern blot analysis of a panel of RNAs from normal human tissues using the PSD cDNA as probe revealed the presence of three different tissue-specific transcripts of approximately 4.3, 2.3, and 1.8 kb, the longest of which is expressed only in brain. Our data suggest that the PSD gene may code for a protein related to a recently identified protein family containing both the Sec7 and the PH domains thought to be involved in signaling transduction processes. PMID- 9417913 TI - Structural organization of the human flavin-containing monooxygenase 3 gene (FMO3), the favored candidate for fish-odor syndrome, determined directly from genomic DNA. AB - The inherited metabolic disorder trimethylaminuria (fish-odor syndrome) is associated with defective hepatic N-oxidation of dietary-derived trimethylamine catalyzed by flavin-containing monooxygenase (FMO). As FMO3 encodes the major form of FMO expressed in adult human liver, it represents the best candidate gene for the disorder. The structural organization of FMO3 was determined by sequencing the products of exon-to-exon and vectorette PCR, the latter through the use of vectorette libraries constructed directly from genomic DNA. The gene contains one noncoding and eight coding exons. Knowledge of the exon/intron organization of the human FMO3 gene enabled each of the coding exons of the gene, together with their associated flanking intron sequences, to be amplified from genomic DNA and will thus facilitate the identification of mutations in FMO3 in families affected with fish-odor syndrome. PMID- 9417914 TI - Cloning and characterization of a novel rho-type GTPase-activating protein gene (ARHGAP6) from the critical region for microphthalmia with linear skin defects. AB - Microphthalmia with linear skin defects syndrome (MLS) is an X-linked dominant, male-lethal disorder associated with chromosomal rearrangements that result in deletions of the distal short arm of the X chromosome. In an effort to isolate expressed sequences from the 500-kb MLS critical region in Xp22.3, exons were trapped from 14 overlapping cosmids. Using exon connection followed by cDNA library screening, we identified a 2.4-kb contig of cDNA library screening 170 kb of genomic sequence in the MLS deletion region. Northern analysis of this cDNA detected a prominent approximately 4.2-kb transcript and a less abundant approximately 6-kb transcript in all tissues examined, with additional transcripts in skeletal muscle. Sequence analysis revealed a coding region of 601 amino acids contained in 12 exons, with a splice variant isoform of 495 amino acids. The predicted protein sequence of the gene, named ARHGAP6, contains homology to the GTPase-activating (GAP) domain of the rhoGAP family of proteins, which has been implicated in the regulation of actin polymerization at the plasma membrane in several cellular processes. The possible role of the ARHGAP6 protein in the pathogenesis of MLS is discussed. PMID- 9417915 TI - Cloning and genomic organization of the human transforming growth factor-beta type I receptor gene. AB - Transforming growth factor-beta (TGF beta) regulates cell cycle progression by a unique signaling mechanism that involves its binding to the type II (T beta R-II) TGF beta receptor and activation of type I (T beta R-I). Both are transmembrane serine-threonine receptor kinases. As various types of human tumor cells are often refractory to TGF beta-mediated cell cycle arrest, it is likely that the T beta R-I receptor is inactivated in many of these cases. We determined the intron exon organization of the TGFBR1 gene. We report here that this gene is approximately 31 kb in length and consists of nine exons. The organization of the segment of the TGFBR1 gene that encodes the C-terminal portion of the serine threonine kinase domain appears to be highly conserved between members of the R-I gene family. This information should facilitate and expedite the structural analysis of TGFBR1 in human tumors and possibly other disease states. PMID- 9417916 TI - Isolation of a pancreas-specific gene located on human chromosome 14q31: expression analysis in human pancreatic ductal carcinomas. AB - We have isolated a novel cDNA (SEL1L) that shows sequence similarities to SEL-1, a gene identified as an extragenic suppressor of the lin-12 hypomorphic mutant from Caenorhabditis elegans (7, 8). SEL1L exhibits a tissue-specific pattern of expression: a single poly(A)+ RNA species of 7.5 kb is abundantly expressed only in the pancreas of healthy individuals, whereas low to undetectable levels are observed in other adult and in some fetal tissues. Somatic hybrid panel and fluorescence in situ hybridization positioned this gene in the q31 band of human chromosome 14. The tissue-specific expression of this gene induced us to study its role in human pancreatic carcinomas. Our analysis revealed that 17% of adenocarcinomas of the pancreas did not express SEL1L to a detectable level; however, no gross genomic alterations were apparent in the few hundred kilobases of the relevant region. PMID- 9417917 TI - Neuropeptide Y receptor genes mapped in human and mouse: receptors with high affinity for pancreatic polypeptide are not clustered with receptors specific for neuropeptide Y and peptide YY. AB - Ppyr1, Npy5r, and Npy6r, the genes encoding mouse type 4, type 5, and type 6 members of the neuropeptide Y receptor family, have been mapped by interspecific backcross analysis to conserved linkage groups on mouse Chr 14, Chr 8, and Chr 18, respectively. The human genes, PPYR1 and NPY5R, have been localized to chromosomes 10q and 4q, respectively, by analysis of a panel of rodent-human somatic cell hybrids and yeast artificial chromosomes. These studies complete the mapping of the cloned NPY receptor subtypes in human and mouse and, together with previous studies, establish that the genes encoding receptors with high affinity for pancreatic polypeptide are not clustered with the genes encoding receptors specific for neuropeptide Y and peptide YY. The physical association of these receptor genes correlates with ligand-binding properties, rather than sequence identity, and suggests a complex evolutionary relationship. PMID- 9417918 TI - Limatin (LIMAB1), an actin-binding LIM protein, maps to mouse chromosome 19 and human chromosome 10q25, a region frequently deleted in human cancers. AB - LIM domains, found in over 60 proteins, play key roles in the regulation of developmental pathways. They were first identified as cysteine-rich motifs found in the three proteins Lin-11, Isl-1, and Mec-3. LIM proteins frequently contain DNA-binding homeodomains, allowing these proteins to activate transcription. LIM domains also function as protein-binding interfaces, mediating specific protein protein interactions. Limatin is a novel LIM protein that binds to actin filaments via a domain that is homologous to erythrocyte dematin. Here we report the murine and human chromosomal localizations of limatin (LIMAB1). Limatin was mapped to mouse Chromosome 19 by restriction fragment length polymorphism analysis and to human chromosome region 10q25 by fluorescence in situ hybridization. Radiation hybrid mapping placed LIMAB1 in a 37-cR interval between markers D10S554 and D10S2390. Interestingly, 10q25 is a region of frequent loss of heterozygosity in human tumors, thus identifying limatin as a candidate tumor suppressor gene. PMID- 9417919 TI - Isolation of human and fission yeast homologues of the budding yeast origin recognition complex subunit ORC5: human homologue (ORC5L) maps to 7q22. AB - Orc5p is a subunit of the origin recognition complex in the budding yeast Saccharomyces cerevisiae, which has been shown to play a critical role in both chromosomal DNA replication and transcriptional silencing. We have cloned cDNAs from both human and fission yeast Schizosaccharomyces pombe that encode proteins homologous to the budding yeast and Drosophila Orc5p. Human Orc5p showed 35.1, 22.3, and 19.4% identity to the Drosophila, S. pombe, and S. cerevisiae Orc5p, respectively. We have localized the human ORC5 gene (ORC5L) to chromosome 7 using Southern and PCR analysis of DNA isolated from a panel of human/rodent somatic cell hybrids and mapped the gene locus to 7q22 using fluorescence in situ hybridization. We have identified a YAC clone that contains human ORC5L and maps to chromosome band 7q22.1. We have identified the S. pombe ORC5 gene and located it in a cosmid mapped on chromosome II. PMID- 9417920 TI - The order and transcriptional orientation of the human COL13A1 and P4HA genes on chromosome 10 long arm determined by high-resolution FISH. AB - The genes for type XIII collagen (COL13A1) and prolyl 4-hydroxylase (P4HA) were previously assigned to human chromosome 10q by radioactive in situ hybridization. Here we have applied fluorescence in situ hybridization combined with targets representing different levels of resolution to determine, first, the order of these genes along chromosome 10; second, their transcriptional orientation; and third, the distance between these genes. The order along the chromosome was determined to be centromere-COL13A1-P4HA-telomere using mechanically stretched chromosomes. By combining the data from stretched chromosomes and interphase nuclei, we found that the transcriptional orientation were tail to tail (COL13A1 3'-3' P4HA). The distance between these genes was measured by fiber FISH to be approximately 550 kb. PMID- 9417921 TI - Fine mapping of 39 ESTs on human chromosome 6p23-p25. AB - Loci conferring susceptibility to schizophrenia, coeliac disease, and orofacial clefting have been assigned to the 6p23-p25 region of human chromosome 6. To facilitate the identification of candidate genes we have sublocalized and ordered 39 ESTs assigned to this interval by radiation hybrid mapping. This was achieved by generating PAC contigs containing the ESTs, genetic markers, and random STSs. For full integration into previously published data a single YAC contig spanning 6p23-p25 was used to unambiguously order the PAC contigs and ESTs along the chromosome. The majority of the ESTs (31/39) were positioned in the 6p23-p24 interval at the proximal half of the map, and of these 8 are located within a single PAC clone. The order of known genes in this region is cen-CD83-ZNF40-EDN1 (GCNT2, CAPZB)-TFAP2-BMP6-DSP-tel. PMID- 9417922 TI - The homozygous complete hydatidiform mole: a unique resource for genome studies. AB - The most frequent type of complete hydatidiform mole is a 46, XX homozygote formed by the fertilization of an empty ovum by a single haploid sperm that later duplicates its chromosomes to give a diploid tumor. The homozygous nature of these complete hydatidiform moles makes them unique resources for human genome studies. They can serve as homozygous controls in the development of single nucleotide polymorphism (SNP) markers and provide a way to obtain long-range haplotypes that are useful in population studies. The use of a homozygous control makes it possible to estimate the allele frequencies of the SNP markers in any population by sequencing pooled DNA samples. In this report, we present evidence of homozygosity of a complete hydatidiform mole using 20 diallelic markers distributed across the genome. Furthermore, its usefulness as a homozygous control in SNP development and as a resource for long-range haplotype determination is demonstrated using 11 newly discovered loci in the BRCA2 region on chromosome 13q12-q13. PMID- 9417923 TI - Localization of BRRN1, the human homologue of Drosophila barr, to 2q11.2. PMID- 9417924 TI - Localization of the human HIP1 gene close to the elastin (ELN) locus on 7q11.23. PMID- 9417926 TI - Crystal structure of herbicide-detoxifying maize glutathione S-transferase-I in complex with lactoylglutathione: evidence for an induced-fit mechanism. AB - Glutathione S-transferases (GSTs) -I and -III are involved in herbicide metabolism in maize and have been intensively studied. Starting with plant tissue from Zea mays var. mutin recombinant GST-I was prepared by heterologous expression in Escherichia coli. The enzyme was crystallized in the presence of lactoylglutathione, a ligand formerly never observed in a GST structure and known as an intermediate of the pharmacologically relevant glyoxalase system. The crystal structure of GST-I has been determined at 2.5 A resolution and exhibits the GST-typical dimer of two identical subunits, each consisting of 214 residues. Compared with other plant GSTs the three-dimensional structure of GST-I primarily shows structural differences in the hydrophobic substrate binding site, the linker segment and the C-terminal region. Furthermore, a comparison of the ligand bound GST-I structure with the apo structure of GST-III indicates the movement of a ten-residue loop upon binding of the ligand to the active site. This is the first structure-based evidence for an induced fit mechanism of glutathione S transferases, which has previously been postulated for class pi enzymes. Together with GST-III, GST-I may explain herbicide resistance and selectivity in maize as well as in other agronomic relevant crops. PMID- 9417925 TI - Importance of minor groove binding zinc fingers within the transcription factor IIIA-DNA complex. AB - The gene-specific transcription factor IIIA (TFIIIA) binds to the internal promoter element of the 5 S rRNA gene through nine zinc fingers which make specific DNA contacts. Seven of the nine TFIIIA zinc fingers participate in major groove DNA contacts while two fingers, 4 and 6, have been proposed to bind in or across the minor groove. Pyrrole-imidazole polyamides are minor groove binding ligands that recognize predetermined DNA sequences with affinity and specificity comparable to natural DNA-binding proteins. We have examined the DNA binding activity of nine finger TFIIIA and shorter recombinant analogs in the presence of polyamides that bind six base-pair sequences (Kd = 0.03 to 1.7 nM) in the minor groove of the binding site for zinc finger 4. DNase I footprint titrations demonstrate that the polyamides and a recombinant protein containing the three amino-terminal zinc fingers of TFIIIA (zf1-3) co-occupy the TFIIIA binding site, in agreement with the known location of zf1-3 in the major groove. In contrast, the polyamides block the specific interaction of TFIIIA or zf1-4 with the 5 S RNA gene, supporting a model for minor groove occupancy by zinc finger 4. Minor groove binding polyamides targeted to specific DNA sequences may provide a novel chemical approach to probing multidomain protein-DNA interactions. PMID- 9417927 TI - Clusters of nucleosomes containing chromosomal protein HMG-17 in chromatin. AB - Chromosomal proteins HMG-14 and HMG-17 are nucleosome binding proteins which can function as architectural elements to alter the structure of the chromatin fiber and enhance transcription from chromatin templates. Here we study the spatial organization of these HMG proteins in the nucleus and the distribution of nucleosomes containing HMG-17 in the chromatin fiber. By confocal immunofluorescence microscopy we find that HMG-14/17 proteins are clustered into foci containing either HMG-14 or HMG-17. These results suggest that HMG-14/17 proteins segregate into distinct nuclear domains. Indeed, immunofractionation of defined length oligonucleosomes, with affinity pure antibodies to HMG-17, indicates that oligonucleosomes containing HMG-17 are devoid of HMG-14. Quantitative analysis indicates that in cellular chromatin nucleosomes containing HMG-17 are clustered. The average size of the cluster is six contiguous HMG-17 containing nucleosomes. The nucleosomes in this cluster contain either two or zero molecules of HMG-17 and a complete set of four core histones. We suggest that HMG-14/17 proteins modify the nucleosomal organization of the 30 nm chromatin fiber, to unfold the higher order chromatin structure and facilitate access to the underlying DNA sequence. Clustering of architectural elements, such as HMG proteins and linker histone subtypes into distinct domains, may lead to structural and functional heterogeneity along the chromatin fiber. PMID- 9417928 TI - Specific DNA recognition by the Aspergillus nidulans three zinc finger transcription factor PacC. AB - The three zinc fingers of PacC, the transcription factor mediating pH regulation in Aspergillus nidulans, are necessary and sufficient to recognise specifically the target ipnA2 site. Missing nucleoside footprints confirmed the core target (double-stranded) hexanucleotide 5'-GCCAAG-3'. Any base substitution resulted in substantial or complete loss of binding, excepting A5 (partially replaceable by G). A T preceding the hexanucleotide enhanced binding. Interference footprinting indicates that the four Gs and A4 participate in specific contacts and that five pyrimidines are essential for binding. The size of the target sequence and the amino acid sequence of finger 1 suggested that its probe helix would not participate in base-specific contacts. Using site-directed mutagenesis and analogy to GLI, we propose that finger 1 crucially interacts with finger 2, a pair of conserved Trp residues in the Cys knuckles contacting hydrophobically. Finger 2 would also participate in extensive base contacts with the 5' moiety of the hexanucleotide. The specificity mutation Lys159Gln shows that finger 3 binds the 3' moiety of the hexanucleotide. Replacement of residues in positions +3 (His128Asn) and +2 (Gln155Lys) of the reading helices of fingers 2 and 3, respectively, prevented binding. Our biochemical and molecular data plus modelling using previously determined zinc finger-DNA complexes, predict specific contacts of fingers 2 and 3 to ipnA2. Our data indicate compact organisation of the PacC-ipnA2 complex (with nearly every base involved in specific contacts), illustrate the binding versatility of zinc finger domains and should facilitate analysis of other PacC family members, including Saccharomyces cerevisiae RIM1. PMID- 9417929 TI - Structural features in the 3' external transcribed spacer affecting intragenic processing of yeast rRNA. AB - A highly conserved extended hairpin structure in the 3' external transcribed spacer (3' ETS) region of nascent eukaryotic rRNA transcripts is essential for the maturation of the large ribosomal subunit RNAs (5.8 S and 25 to 28 S rRNAs). Systematic changes were introduced into this structure by PCR-mediated mutagenesis and the mutant rDNAs were expressed in vivo to determine the structural features that are essential for rRNA maturation. Changes in the lower half of the stem or the large loop at the end had little or no effect on the maturation of either the 5.8 S or 25 S rRNA, but changes that disrupted secondary structure in the upper half of this stem had equal and dramatic effects on both RNAs. When the RNA stem was incubated with a cellular protein extract, gel retardation studies indicated that the stem forms a ribonucleoprotein complex, and a comparison with mutant RNA indicated that protein binding could be compromised by changes that were critical for rRNA maturation. Sequence comparisons with other spacer regions as well as snRNAs reveal some structural analogy, which, when taken together with the mutational studies, raise the possibility that this hairpin functions during RNA processing in a manner that may be analogous with that of free snRNPs. PMID- 9417930 TI - Defining functional regions of the IS903 transposase. AB - The insertion sequence IS903 encodes a 307 amino acid residue protein, transposase, that is essential for transposition. It is a multi-functional DNA binding protein that specifically recognizes the 18 bp inverted repeats at the ends of the element and also recognizes DNa non-specifically when it captures a target site. In addition, transposase performs catalytic functions when it mediates the cleavage and religation steps of transposition. We have carried out deletion and mutational analyses to define functional domains of the transposase protein. The deletion studies delineate a 99 residue region of the protein (residues 31 to 129) that specifies binding to the inverted repeat. A slightly larger maltose-binding protein-transposase fusion that includes residues 22 to 139 (Tnp 22-139) binds as efficiently and with the same specificity as the full length transposase protein. Tnp 22-139 also induces a DNA bend similar to that of the wild-type protein, and so we conclude that all binding and bending specificity is contained within the N-terminal domain of the protein. Unlike full length transposase, Tnp 22-139 forms additional higher-order complexes in band shift gels suggesting that the deletion has exposed a surface(s) capable of participating in protein-protein interactions. Six highly conserved residues in the C-terminal portion of the protein were mutated to alanine. Each mutant protein was binding-proficient but defective in transposition. The phenotype of these substitutions, and their alignment with residues shown to abolish catalysis of other transposases and integrases, suggest that these are residues responsible for catalytic steps in transposition of IS903; we believe three of these residues comprise the DDE motif, conserved in transposases and integrases. Our data are consistent with IS903 transposase being composed of two domains: an N-terminal domain primarily involved in DNA binding and a C-terminal domain that is involved in catalysis. PMID- 9417931 TI - Major identity determinants for enzymatic formation of ribothymidine and pseudouridine in the T psi-loop of yeast tRNAs. AB - Almost all transfer RNA molecules sequenced so far contain two universal modified nucleosides at positions 54 and 55, respectively: ribothymidine (T54) and pseudouridine (psi 55). To identify the tRNA elements recognized by tRNA:m5uridine-54 methyltransferase and tRNA:pseudouridine-55 synthase from the yeast Saccharomyces cerevisiae, a set of 43 yeast tRNA(Asp) mutants were used. Some variants contained point mutations, while the others included progressive reductions in size down to a tRNA minisubstrate consisting of the T psi-loop with only one G.C base-pair as stem (9-mer). All substrates (full-sized tRNA(Asp) and various minihelices) were produced in vitro by T7 transcription and tested using yeast extract (S100) as a source of enzymatic activities and S-adenosyl-L methionine as a methyl donor. The results indicate that the minimal substrate for enzymatic formation of psi 55 is a stem/loop structure with only four G.C base pairs in the stem, while a longer stem is required for efficient T54 formation. None of the conserved nucleotides (G53, C56, A58 and C61) and U54 for psi 55 or U55 for T54 formation can be replaced by any of the other three canonical nucleotides. Yeast tRNA:m5uridine-54 methyltransferase additionally requires the presence of a pyrimidine-60 in the loop. Interestingly, in a tRNA(Asp) variant in which the T psi-loop was permuted with the anticodon-loop, the new U32 and U33 residues derived from the T psi-loop were quantitatively converted to T32 and psi 33, respectively. Structural mapping of this variant with ethylnitrosourea confirmed that the intrinsic characteristic structure of the T psi-loop was conserved upon permutation and that the displaced anticodon-loop did not acquire a T psi-loop structure. These results demonstrate that a local conformation rather than the exact location of the U-U sequence within the tRNA architecture is the important identity determinant for recognition by yeast tRNA:m5uridine-54 methyltransferase and tRNA:pseudouridine-55 synthase. PMID- 9417932 TI - Essential structures of a self-aminoacylating RNA. AB - Comparison of six independent self-aminoacylating RNAs derived from selection amplification, as well as deletion, addition, substitution, fragmentation of one particular RNA, are used to analyze the requirements for the RNA-catalyzed aminoacylation. All elements required for catalysis by one RNA family: sequence at the 3' acceptor end, calcium and magnesium sites, as well as the Phe-AMP substrate site and the essential 5' triphosphate terminus, are closely grouped near a bihelix junction in the parental molecule. All elements of the active center for aminoacyl transfer can therefore be captured by a peripherally-deleted helix junction RNA, defining a much smaller 43 nucleotide ribozyme, of which only 17 nucleotides were initially randomized. It appears that a complex RNA active center can be assembled by specifying unexpectedly few nucleotides, perhaps with a critical contribution from an essential calcium ion. PMID- 9417933 TI - Sequence profiles of immunoglobulin and immunoglobulin-like domains. AB - Immunoglobulins (Ig) are highly modular proteins, consisting of variable and constant domains, which have clear, conserved sequence patterns. These sequence patterns have allowed T-cell receptor (TCR) and major histocompatibility complex (MHC) molecule domains, as well as some cell adhesion, cell surface receptor and muscle protein domains, to be identified as forming a superfamily of related proteins together with the Ig-domains. The domains of these proteins have been grouped into four sets: variable (V-set), constant-1 (C1-set), constant-2 (C2 set) and intermediate (I-set). X-ray and NMR studies have shown that these domains form a Greek-key beta-sandwich structure with the sets differing in the number of strands in the beta-sheets as well as in their sequence patterns. The conserved sequence elements in the major sets of Ig and Ig-like molecules have previously been reported as general sequence profiles. This work examines the variability within these sets. Detailed sequence profiles and consensus sequences for these sets and groups have been constructed and a novel form of presentation has been developed to overcome some of the drawbacks of current methods of presenting consensus sequences. The profiles that were constructed allow a comparison of the similarities and differences among the sets of Ig and Ig-like sequences and provide a means by which sequences can be tested for compatibility with Ig-like sequence motifs. As well, the sequence separations of the main residues in the characteristic "pin" structure of Ig-like molecules were examined for variation among the groups. From the profiles constructed here, measures of the degree of conservation within the groups of molecules were determined. These measures were used to assist in a reconsideration of possible evolutionary pathways between the major structural groups of the Ig-superfamily. PMID- 9417934 TI - The Escherichia coli dnaA gene: four functional domains. AB - The Escherichia coli DnaA protein is a sequence-specific DNA binding protein that promotes the initiation of replication of the bacterial chromosome, and of several plasmids including pSC101. Twenty-eight novel missense mutations of the E. coli dnaA gene were isolated by selecting for their inability to replicate a derivative of pSC101 when contained in a lambda vector. Characterization of these as well as seven novel nonsense mutations and one in-frame deletion mutation are described here. Results suggest that E. coli DnaA protein contains four functional domains. Mutations that affect residues in the P-loop or Walker A motif thought to be involved in ATP binding identify one domain. The second domain maps to a region near the C terminus and is involved in DNA binding. The function of the third domain that maps near the N terminus is unknown but may be involved in the ability of DnaA protein to oligomerize. Two alleles encoding different truncated gene products retained the ability to promote replication from the pSC101 origin but not oriC, identifying a fourth domain dispensable for replication of pSC101 but essential for replication from the bacterial chromosomal origin, oriC. PMID- 9417935 TI - A structural census of genomes: comparing bacterial, eukaryotic, and archaeal genomes in terms of protein structure. AB - Representative genomes from each of the three kingdoms of life are compared in terms of protein structure, in particular, those of Haemophilus influenzae (a bacteria), Methanococcus jannaschii (an archaeon), and yeast (a eukaryote). The comparison is in the form of a census (or comprehensive accounting) of the relative occurrence of secondary and tertiary structures in the genomes, which particular emphasis on patterns of supersecondary structure. Comparison of secondary structure shows that the three genomes have nearly the same overall secondary-structure content, although they differ markedly in amino acid composition. Comparison of super-secondary structure, using a novel "frequent words" approach, shows that yeast has a preponderance of consecutive strands (e.g. beta-beta-beta patterns), Haemophilus, consecutive helices (alpha-alpha alpha), and Methanococcus, alternating helix-strand structures (beta-alpha-beta). Yeast also has significantly more helical membrane proteins than the other two genomes, with most of the differences concentrated in proteins containing two transmembrane segments. Comparison of tertiary structure (by sequence matching and domain-level clustering) highlights the substantial duplication in each genome (approximately 30% to 50%), with the degree of duplication following similar patterns in all three. Many sequence families are shared among the genomes, with the degree of overlap between any two genomes being roughly similar. In total, the three genomes contain 148 of the approximately 300 known protein folds. Forty-five of these 148 that are present in all three genomes are especially enriched in mixed super-secondary structures (alpha/beta). Moreover, the five most common of these 45 (the "top-5") have a remarkably similar super secondary structure architecture, containing a central sheet of parallel strands with helices packed onto at least one face and beta-alpha-beta connections between adjacent strands. These most basic molecular parts, which, presumably, were present in the last common ancestor to the three Kingdoms, include the TIM barrel, Rossmann, flavodoxin, thiamin-binding, and P-loop-hydrolase folds. PMID- 9417936 TI - Cloning, sequencing, crystallization and X-ray structure of glutathione S transferase-III from Zea mays var. mutin: a leading enzyme in detoxification of maize herbicides. AB - Glutathione S-transferases (GSTs) are enzymes that inactivate toxic compounds by conjugation with glutathione and are involved in resistance towards drugs, antibiotics, insecticides and herbicides. Their ability to confer herbicide tolerance in plants provides a tool to control weeds in a wide variety of agronomic crops. GST-III was prepared from Zea mays var. mutin and its amino acid sequence was determined from two sets of peptides obtained by cleavage with endoprotease Asp-N and with trypsin, respectively. Recombinant GST-III was prepared by extraction of mRNA from plant tissue, transcription into cDNA, amplification by PCR and expression. It was crystallized and the crystal structure of the unligated form was determined at 2.2 A resolution. The enzyme forms a GST-typical dimer with one subunit consisting of 220 residues. Each subunit is formed of two distinct domains, an N-terminal domain consisting of a beta-sheet flanked by two helices, and a C-terminal domain, entirely helical. The dimeric molecule is globular with a large cleft between the two subunits. The amino acid sequence of GST-III and its cDNA sequence determined here show differences from sequences published earlier. PMID- 9417937 TI - Contrasting roles for symmetrically disposed beta-turns in the folding of a small protein. AB - To investigate the role of turns in protein folding, we have characterized the effects of combinatorial and site-directed mutations in the two beta-turns of peptostreptococcal protein L on folding thermodynamics and kinetics. Sequences of folded variants recovered from combinatorial libraries using a phase display selection method were considerably more variable in the second turn than in the first turn. These combinatorial mutants as well as strategically placed point mutants in the two turns had a similar range of thermodynamic stabilities, but strikingly different folding kinetics. A glycine to alanine substitution in the second beta-turn increased the rate of unfolding more than tenfold but had little effect on the rate of folding, while mutation of a symmetrically disposed glycine residue in the first turn had little effect on unfolding but slowed the rate of folding nearly tenfold. These results demonstrate that the role of beta-turns in protein folding is strongly context-dependent, and suggests that the first turn is formed and the second turn disrupted in the folding transition state. PMID- 9417938 TI - Identification of critical IgG binding epitopes on the neonatal Fc receptor. AB - The neonatal Fc receptor (FcRn) binds maternal immunoglobulin G (IgG) during the acquisition of passive immunity by the fetus or newborn. FcRn also binds IgG and returns it to the bloodstream, thus protecting IgG from a default degradative pathway. Biosensor assays have been used to characterize the interaction of a soluble form of rat FcRn with IgG, and demonstrate that FcRn dimerization and immobilization are necessary to reproduce in vivo binding characteristics. Here, we report the identification of several FcRn amino acid substitutions that disrupt its affinity for IgG and examine the effect of alteration of residues at the FcRn dimer interface. The role of these amino acids is discussed in the context of the previously reported structures of rat FcRn and a complex of FcRn with the Fc portion of IgG. PMID- 9417939 TI - Structure of dihydrodipicolinate synthase of Nicotiana sylvestris reveals novel quaternary structure. AB - DHDPS is the first enzyme unique to the lysine biosynthetic pathway in plants and bacteria and catalyses the formation of (4S)-4-hydroxy-2,3,4,5-tetrahydro-(2S) dipicolinic acid. It is feedback-regulated in plants by L-lysine. The crystal structure of Nicotiana sylvestris DHDPS with and without inhibitory lysine bound to the enzyme has been solved to a resolution of 2.8 A. The molecule is a homotetramer composed of a dimer of dimers. Comparison with the structure of Escherichia coli DHDPS showed a novel quaternary structure by a profound rearrangement of the dimers forming the tetramer. The crystal structure of the enzyme in the presence of L-lysine revealed substantial changes. These changes together with the novel quaternary structure provide a structural basis for the strong inhibition of plant DHDPS enzymes by L-lysine. PMID- 9417940 TI - The three-dimensional structures of a polysaccharide binding antibody to Cryptococcus neoformans and its complex with a peptide from a phage display library: implications for the identification of peptide mimotopes. AB - The three-dimensional structure of 2H1, a protective monoclonal antibody to Cryptococcus neoformans, has been solved at 2.4 A resolution, in both its unbound form and in complex with the 12 amino acid residue peptide PA1 (GLQYTPSWMLVG). PA1 was previously identified as a potential mimotope of the cryptococcal capsular polysaccharide by screening of a phage display peptide library. Peptide binding is associated with only minor rearrangements of some side-chains and a small shift in the H2 loop of the antibody. The peptide assumes a tightly coiled conformation consisting of one inverse gamma-turn and one type II beta-turn that serves to place the entire peptide motif, consisting of ThrP5, ProP6, TrpP8, MetP9 and LeuP10, into a depression in the antibody combining site. A small number of H-bonds between peptide and antibody contribute to the affinity and specificity. Poor steric complementarity between PA1 and the antibody heavy chain along with the fact that the majority of the interactions between 2H1 and PA1 involve van der Waals interactions with the light chain may explain why this peptide acts as only a partial mimotope of the capsular polysaccharide epitope. PMID- 9417941 TI - The 2.4 A resolution crystal structure of boar seminal plasma PSP-I/PSP-II: a zona pellucida-binding glycoprotein heterodimer of the spermadhesin family built by a CUB domain architecture. AB - The crystal structure of porcine seminal plasma spermadhesin PSP-I/PSP-II heterodimer has been determined in two crystal forms by multiple isomorphous replacement in an hexagonal crystal (space group P6(1)22) and molecular replacement in a trigonal crystal of space group P3(2)21. The crystal structure has been refined at 2.4 A resolution to an R-factor of 20.0% (Rfree = 25.9%) for 14,809 independent reflections with intensities greater than 2 sigma (I), with root-mean-square deviations of 0.009 A and 1.657 degrees from ideal bond lengths and bond angles, respectively. The final model includes 1688 non-hydrogen protein atoms of 221 amino acids and 79 water molecules. PSP-I/PSP-II represents the first crystal structure of a mammalian zona pellucida-binding protein. PSP-II displays a putative carbohydrate-recognition site located around its Asn50. This region shares structural features with sugar binding sites of known lectin structures of the leguminous and galectin families. PSP-I and PSP-II are N glycosylated at asparagine residues 50 and 98, respectively, and show site heterogeneity. Only the innermost N-acetylglucosamine of PSP-I is defined in the crystal structure. Both subunits of the PSP-I/PSP-II heterodimer are built by a single CUB domain architecture. The CUB domain displays a novel fold, which consists of a compact ellipsoidal beta-sandwich structure (42 A x 27 A x 23 A) organized into two 5-stranded beta-sheets. Each sheet contains parallel and antiparallel beta-strands. Two disulphide bridges, which are conserved in all spermadhesin molecules and many CUB domains, crosslink loop LA and strand beta 4 and loops LE and LG, respectively, at opposite edges of the same face of the domain. The four highly conserved aromatic residues and 15 out of 17 invariant hydrophobic residues, which define the CUB domain signature, display an interior location, suggesting that this hydrophobic core may be essential for maintaining the overall folding of the domain. Most of the hydrophobic core residue characteristics are conserved in the jellyroll topology of certain icosahedral virus capsid proteins, indicating that the CUB domain and the viral proteins share a minimal structural core. PMID- 9417942 TI - Crystal structure of acidic seminal fluid protein (aSFP) at 1.9 A resolution: a bovine polypeptide of the spermadhesin family. AB - We report the three-dimensional crystal structure of acidic seminal fluid protein (aSFP), a 12.9 kDa polypeptide of the spermadhesin family isolated from bovine seminal plasma, solved by the multiple isomorphous replacement method and refined with data to 1.9 A resolution with a final R-factor of 17.3%. aSFP is built by a single CUB domain architecture, a 100 to 110 amino-acid-residue extracellular module found in 16 functionally diverse proteins. The structure of aSFP reveals that the CUB domain displays a beta-sandwich topology organised into two 5 stranded beta-sheets, each of which contain two parallel and four antiparallel strands. The structure of aSFP is almost identical to that of porcine spermadhesins PSP-I and PSP-II, which in turn show limited structural similarity with jellyroll topologies of certain virus capsid proteins. Essentially, topologically conserved residues in these proteins are those internal amino acids forming the hydrophobic core of the CUB and the jellyroll domains, suggesting their importance in maintaining the integrity of these protein folds. On the other hand, the structure of aSFP shows structural features that are unique to this protein and which may provide a structural ground for understanding the distinct biological properties of different members of the spermadhesin protein family. PMID- 9417943 TI - The three-dimensional high resolution structure of human interferon alpha-2a determined by heteronuclear NMR spectroscopy in solution. AB - The solution structure of recombinant human interferon alpha-2a (Roferon-A) has been determined by multidimensional heteronuclear NMR spectroscopy. The calculations using simulated annealing produced a family of 24 convergent structures which satisfy the experimental restraints comprising 1541 NOE-derived inter-proton distances, 187 dihedral restraints, 66 pairs of hydrogen bond restraints, and six upper and lower limits for two disulfide bridges. The fractional labeling of methyl groups allowed their direct and unambiguous stereospecific assignment which proved to be essential for obtaining a high resolution of the structures. A best fit superposition of residues 10 to 47, 50 to 101 and 111 to 157 gives an rms deviation of 0.62 A for the backbone heavy atoms and 1.39 A for all heavy atoms of these segments. The dominant feature of the structure is a cluster of five alpha-helices, four of which are arranged to form a left-handed helix bundle with an up-up-down-down topology and two over hand connections. The interpretation of heteronuclear 15N-?1H? NOE data shows the co-existence of flexible regions within an otherwise rigid framework of the protein. Four stretches of pronounced flexibility can be located: Cys1-Ser8, Gly44-Ala50, Ile100-Lys112, and Ser160-Glu165. Among the structurally related four-helical bundle cytokines, the structure of IFN alpha-2a is most similar to that of human interferon alpha-2b and murine interferon-beta. From this structural information and mutagenesis data, areas on the surface of the protein are identified which seem to be important in receptor interactions. PMID- 9417944 TI - Disruption of an ionic network leads to accelerated thermal denaturation of D glyceraldehyde-3-phosphate dehydrogenase from the hyperthermophilic bacterium Thermotoga maritima. AB - The role of an ionic network of four charged amino acid side-chains in the thermostability of the enzyme D-glyceraldehyde-3-phosphate dehydrogenase from the hyperthermophilic bacterium Thermotoga maritima (TmGAPDH) has been assessed by site-directed mutagenesis, replacing the central residue of the ionic network, arginine 20, by either alanine (R20A) or asparagine (R20N). The purified mutant enzymes display no differences to the wild-type enzyme regarding spectroscopic properties and enzymatic activity. However, denaturation kinetics reveal that the resistance towards thermal denaturation is strongly diminished in the mutant enzymes. This is reflected by a decrease in free energy of activation for thermal unfolding of about 4 kJ/mol at 100 degrees C and a shift of temperature of half denaturation after one hour incubation from 96 to 89 degrees C for both mutant enzymes. Due to a large decrease in activation enthalpy, the effects of the mutations are temperature dependent and become even more significant at the physiological temperature of Thermotoga maritima (approximately 80 degrees C). The importance of the arginine 20 side-chain for kinetic thermal stability is plausible in the light of its key role in the ionic network and the strategic positioning of this ionic network in the context of the overall protein structure. PMID- 9417945 TI - Rotational Analyses of the Laser Induced Fluorescence Excitation Spectra of Jet Cooled CF3O and CF3S AB - The high resolution, rotationally resolved laser induced fluorescence spectra for the A2A1 left and right arrow X2E transitions of CF3O and CF3S were recorded. In addition to the origins, the symmetric vibrational band 301 and the nontotally symmetric bands 501 and 601 were analyzed for CF3O, while for CF3S the symmetric bands 101, 201, and 301, as well as the nontotally symmetric bands 601, 301501, and 301601, were analyzed. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9417946 TI - High Resolution Fourier Transform Far Infrared Spectroscopy of CH3OD AB - The high resolution Fourier transform far infrared spectrum of the torsion rotation band of CH3OD has been recorded in the range 20-350 cm-1 at a resolution of 0.002 cm-1. The spectrum shows splitting of the lines due to strong torsional, rotational, and vibrational interactions in the molecule. Assignments were possible for rotational subbands in the ground torsional state (n = 0) for K values up to 15 and J values of up to 30, for all the symmetry species. In addition, some subbands were also identified which involve torsionally excited states. A total of 63 b-type subband origins, including 6 Q-branch origins obtained from microwave (MW) and millimeter-wave (MMW) studies, were fitted to a semiempirical model. The molecular parameters so determined were able to reproduce the subband origins almost to within experimental uncertainty. The torsional-rotational state-dependent effective molecular parameters and the asymmetry splitting parameters have also been determined. These should prove valuable in the assignment of transitions involving torsionally excited states in the ground vibrational state. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9417947 TI - Ab initio Relativistic CI Calculations of the Spectroscopic Constants and Transition Probabilities for the Low-Lying States of the BiOH/HBiO Isomers AB - Spin-orbit MRD-CI calculations have been carried out for the potential energy surfaces of the seven lowest-lying electronic states of the BiOH molecule by employing relativistic effective core potentials. The HBiO isomer is found to be 4020 cm-1 less stable because of its inability to form multiple Bi-O bands. A bent 3A" BiOH ground state is predicted, which is split into all three of its components by spin-orbit coupling. The calculated X2A"-X1A' splitting is computed to be 5217 cm-1, but the corresponding X3-X2 value is only 29 cm-1. Fink et al. have observed spectral bands which appear with a Te value of 6200 cm-1 which are likely caused by BiOH. Since calculations at the same level for BiF underestimate the observed X2-X1 spin-orbit splitting by 650 cm-1, it appears that the present calculations are consistent with this experimental assignment. A vibrational progression with a 500 cm-1 frequency is also observed and this result fits in well with the computed Bi-O stretch omegae value of 527 cm-1. The calculations also find a relatively large 1Delta splitting (600 cm-1) because of the bent BiOH geometry, with comparatively strong transitions to the X1A' ground state, and it is suggested that the experimental BiOH assignment can be confirmed on this basis. Much stronger transitions to the 1Delta component should also be observed in emission in the 10 000 cm-1 range. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9417948 TI - High Resolution FTIR Study of the nu4 + 2nu6 Rovibrational Band of CH379Br between 4870 and 5030 cm-1 AB - The nu4+/-1 + 2nu60 and nu4-/+1 + 2nu6+/-2 perpendicular components and the nu4+/ 1 + 2nu6+/-2 parallel component of the CH379Br isotopomer have been found and studied in high resolution (0.006 cm-1). Both perpendicular components are linked altogether and also to the nu2 + nu3 + nu4 band by anharmonic resonances. A model taking these anharmonic resonances and l(2,2) resonances into account has been used. It allowed to obtain a r.m.s. deviation of 0.0065 cm-1 in a least squares fit over 1327 lines (46 being zero-weighted) belonging to the three components of nu4 + 2nu6 and to the calculated KDeltaK = -1 sub-band of nu2 + nu3 + nu4 deduced from the corresponding observed sub-band of nu2 + nu3 + nu4 - nu3. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9417949 TI - New Band Systems of the YBr Molecule AB - The thermal emission spectrum of the YBr molecule has been photographed in the region 4130-4750 A at a temperature about 2300degreesC using Saha's vacuum furnace at a dispersion of 3.5 A/mm on the C. Z. Ebert plane grating spectrograph. A total of 123 red-degraded bands have been recorded and assigned to two new systems, namely F-X and G-X, and one previously reported system, viz. E-X. The vibrational constants have been evaluated for these systems. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9417950 TI - Diode-Laser Measurements of He-Broadening Coefficients in the nu6 Band of 12CH3F AB - He-broadening coefficients are measured for 28 lines of 12CH3F in the RQ0 (Ki = 0, DeltaK = 1, DeltaJ = 0) and RQ6 (Ki = 6, DeltaK = 1, DeltaJ = 0) branches of the nu6 band, using a tunable diode laser spectrometer. The collisional widths obtained by fitting Rautian profiles to the measured shapes of the lines are slightly larger than those derived from Voigt profiles. The broadening coefficients for the lines belonging to the same low J transitions are significantly greater in the RQ0 branch than in the RQ6 branch. Moreover, they decrease on the whole with increasing J in the RQ0 branch and tend to increase from J varying from 7 to 10 in the RQ6 branch. These trends may be approximately reproduced by calculations based on a semiclassical impact theory of collisional broadening. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9417951 TI - The Fundamental Vibrations of NC-CC-CN (Dicyanoacetylene) AB - In the infrared spectrum of dicyanoacetylene the rotation vibration bands nu1 - nu9/2nu5 - nu9, nu1 + nu9/2nu5 + nu9 (Fermi resonance), nu2 - nu9, nu2 + nu8, nu4, nu6 - nu9, nu6 + nu8 - nu6, nu4 - nu7, nu4 + nu7, nu4 + nu7 - nu7, nu7 + nu8 - nu7, and 2nu7 + nu8 - nu7 (I and II) have been recorded with high resolution. From these bands the vibrational levels v1 = 1, v2 = 1, v4 = 1, v6 = 1, and v7 = 1 including the corresponding effective rotational constants have been calculated. Different methods for calculating the unperturbed position of the v1 = 1 level have been compared and the unperturbed B1 has been calculated. Additional information on the rotational constants of v8 = 1 and v9 = 1 is also given. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9417952 TI - Line Positions and Strengths of HDO between 6000 and 7700 cm-1 AB - High-resolution spectra of gas sample mixtures of HDO, D2O, and H2O were obtained with a Fourier-transform spectrometer at resolutions of 0.01 and 0.02 cm-1. The spectra were analyzed to obtain line positions and strengths of HDO transitions covering the region from 6000 to 7700 cm-1. The analysis included 2445 lines involving the (101)-(000), (021)-(000), (210)-(000), and (002)-(000) bands; this is the first report of assignments in the (210)-(000) and (002)-(000) bands as well as line strength measurements of any HDO transitions in this spectral interval. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9417953 TI - Testing Lineshape Models: Measurements for v = 1-0 CO Broadened by He and Ar AB - We present measurements of the spectral profiles of many P and R branch lines in CO-He and CO-Ar mixtures, all at 301.5 K. The lineshapes are compared with several semi-classical models and in particular with models that include the speed dependence of both the translational motion and the relaxation of the optical coherence. Using a correlated speed-dependent Galatry model proposed by Ciurylo and Szudy we find an apparent reduction of the Dicke narrowing, amounting to almost 100% in some cases with a strong dependence on the rotational quantum number. However, while the parameters of this and several other models may be adjusted to fit the experimental data within the noise, no single model is free of criticism. Several shortcomings of existing models are discussed. Plots of broadening coefficients versus line number are also given. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9417954 TI - Fourfold Clusters of Rovibrational Energies in H2Po Studied with an ab Initio Potential Energy Function AB - We report here an ab initio investigation of the cluster effect (i.e., the formation of four-member groups of nearly degenerate rotation-vibration energy levels at higher J and Ka values) in the H2Po molecule. The potential energy function has been calculated ab initio for a total of 143 molecular geometries by means of the CCSD(T) method, using an averaged relativistic effective potential for Po in conjunction with a newly optimized basis set. The values of the potential energy function obtained cover the region up to around 5000 cm-1 above the equilibrium energy. On the basis of the ab initio potential, the rotation vibration energies of H2209Po have been calculated with the MORBID (Morse oscillator rigid bender internal dynamics) Hamiltonian and computer program. In particular, we have calculated the rotational energy manifolds for J b2Sigma- (v = 0, 1) transitions in the 1.6 &mgr;m region. Rotational analyses of the doublet bands have been performed. Most of the excited doublet states lie only slightly above or below the well-known B4Pi state. The density of states is fairly high immediately above B4Pi, and characterizations of some of the states in this region involve difficulties. A currently unanalyzed band, centered at 11 820 cm-1, has been preliminarily attributed to the A4Pi --> X4Sigma- (0, 0) transition. Nuclear hyperfine effects are barely detectable in orbitally degenerate states of the doublet manifold at Doppler-limited resolution, while the b2Sigma- state of configuration sigmadelta2 shows partly resolved magnetic hyperfine structure (b = -0.08191 cm-1). Large-scale all-electron CI calculations have been performed on doublet and quartet manifolds of NbO up to 22 000 cm-1. The calculations have been performed in two steps. In the first step, the spin orbit effects within electronic states were not considered. An excellent overall agreement with experimental energies was obtained, except for the d2Deltai state. In the second step, all spin-orbit interactions were included. Also these calculations resulted in a good agreement with the experimental observations. The calculations also predict three hitherto unobserved low-lying states: 4Phi at 8545 cm-1, 2Gamma at 8430 cm-1 and 2Sigma+ at 9648 cm-1. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9417958 TI - The Stimulated Raman Spectrum of Cyanogen AB - The Raman spectrum of cyanogen 12C214N2 has been investigated at nearly Doppler resolution by means of the Stimulated Raman technique. The regions around the Q branches of the nu1 (2330 cm-1) and nu2 (845 cm-1) vibrations have been recorded. Besides the fundamentals, hot bands arising from v5 = 1, v5 = 2, and v4 = 1 have been observed. The spectra have been analyzed, and rotational constants for the excited states have been obtained. Computer simulations of the Raman contours have been carried out as a test of the assignments. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9417959 TI - New N2+ Electronic States in the Region of 23-28 eV AB - Threshold photoelectron spectra of N2+ were measured between 23.4 and 27.6 eV with high resolution and high intensity by using the penetrating field technique and synchrotron radiation. Five vibrational progressions were observed. The first of these progressions was the C2Sigmau+ state. The second progression was identified as the transition to the second state of 2Pig symmetry found by P. Baltzer, M. Larsson, L. Karlsson, B. Wannberg, and M. Carlsson (1992. Phys. Rev. A 46, 5545). The third progression, which was discovered by F. Merkt and P. M. Guyon (1993. J. Chem. Phys. 99, 3400), can be designated as the 2Sigmau- state by comparison with previous theoretical study (E. W. Thulstrup and A. Andersen, 1975. J. Phys. B 8, 965). The fourth and fifth progressions were designated as the 2Deltau and 2(2) Piu states by similar comparison with previous theories. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9417960 TI - Ab Initio Potential Energy Surface and Internal Torsional-Wagging States of Hydroxylamine AB - The two-dimensional potential energy surface describing the interaction of the large-amplitude torsional and wagging motions in hydroxylamine has been determined from ab initio calculations. This surface has been sampled by a large set of grid points from a two-dimensional configuration space spanned by the torsional and wagging coordinates. At each grid point, the geometry optimization has been performed using the second-order Moller-Plesset perturbation theory with the basis set 6-311 + G(2d, p). At the optimized geometry, the single-point calculation of the electronic energy has been carried out using a larger basis set 6-311 + G(3df, 2p). This method was verified to yield the results comparable to those obtained by a direct optimization of the geometry with the basis set 6 311 + G(3df, 2p) which had been used by A. Chung-Phillips and K. A. Jebber (1995. J. Chem. Phys. 102, 7080-7087) to calculate the energies of only three points in the potential energy surface of hydroxylamine. The trans and cis local minima have been found on the determined potential energy surface. The localization features of the torsional-wagging states have been studied by solving the two dimensional Schrodinger equation for the coupled torsional and wagging motions. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9417961 TI - Millimeter-Wave Free Jet Absorption Spectrum of SD Methylthioglycolate: Description of the SH Torsion Double Minimum Potential AB - The free jet absorption millimeter-wave spectrum of methylthioglycolate-SD has been investigated in the 60-78 GHz frequency range. The double-minimum potential associated with the SD group torsion generates a vibrational spacing Delta01 = 8285(5) MHz between the O+ and O- states of the A sublevel. This splitting, compared to the corresponding value of 18 953 MHz for the normal species, suggests a large involvement of the sulfur atom in the motion. A model is given to describe this motion. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9417962 TI - The Rotational Spectrum and Anharmonic Force Field of Chlorine Dioxide, OClO AB - The rotational spectra of O35ClO and O37ClO in their (000), (100), (010), (001), and (020) states have been reinvestigated in selected regions between 130 and 526 GHz. About 800 newly measured lines spanning the quantum numbers 2 200 days). The ontogenic expression of detoxication enzymes was significantly delayed in the marsupial in comparison to eutherians. Adult levels were achieved during the weaning period, suggesting that dietary xenobiotics act as a regulatory mechanism in the developmental expression of these enzymes in the brushtail possum. PMID- 9418015 TI - Effect of carbohydrate moieties on the stability of nuclear glycoproteins from hamster liver. AB - The effect of carbohydrate moieties on the stability of hamster liver nuclear glycoproteins in the course of endogenous proteolysis employing highly specific digoxigenin-labelled lectins, was studied. Whole hamster liver nuclei were autolysed in optimum conditions for the action of nuclear proteinases able to degrade histones as well as non-histone proteins. Incubated samples were electrophoresed on sodium dodecyl sulphate-polyacrylamide gel. Coomassie blue stained gels demonstrated degradation of some proteins in particular after 18 and 24 h incubation. Proteins with molecular weights of about 46, 54 and 76 kD appeared to be resistant to proteolysis. The same location and intensity of bands of glycoproteins on immunoblots from incubated and nonincubated samples of nuclei indicated that oligosaccharide chains protect proteins from degradation. PMID- 9418016 TI - Differential effects of somatostatin analogues on proliferation of murine colonic cancer cells in vitro. AB - The effects of somatostatin analogues octreotide (SMS 201-995), ASS-51 and ASS-52 on [3H]-thymidine incorporation into DNA of the murine colon 38 cancer cells in vitro were investigated. It was found that SMS 201-995 and ASS-51 inhibited the tritiated thymidine incorporation in a dose-dependent manner. In contrast, analogue ASS-52 in spite of a very similar structure to ASS-51, which differed from the latter only by one CH2OH group, was devoid of remarkable antiproliferative activity. These results indicate that slight modification of the molecule of somatostatin analogues may deeply influence their antiproliferative activity. PMID- 9418017 TI - Gap junctions in 9L and C6 glioma cells: correlation with growth characteristics. AB - The relationship between growth properties and gap junctional communication properties of two glioma cell lines, 9L and C6, was compared. Using microinjection of a gap junction permeable fluorescent dye, it was demonstrated that 9L cells were extensively dye coupled through gap junctions, whereas C6 cells had low levels of dye coupling between the cells. More gap junctions were observed between 9L cells than between C6 cells both by electron microscopy and immunofluorescent labelling with specific antibodies. Immunoblotting and immunofluorescence labelling of cellular proteins from 9L and C6 cells also showed differences in the Cx43 connexin contents of these cells, reflecting the differences in their gap junctional communication. Northern blot analyses confirmed that the level of Cx43 mRNA was higher in 9L than in C6 cells. In spite of these differences in gap junctions, the two cell lines did not differ in their growth rates in cultures. These results suggest a lack of direct correlation between gap junctional communication and cell growth regulation in these two glioma cell lines. PMID- 9418018 TI - Antibiotic use in animal agriculture. PMID- 9418019 TI - Pathogenicity of atypical Aeromonas salmonicida in Atlantic salmon compared with protease production. AB - The pathogenicity of extracellular products (ECPs) from 24 atypical Aeromonas salmonicida strains was studied with respect to: lethality in Atlantic salmon, pathogenic effect in muscle, haemolytic activity, cytotoxicity in two fish cell lines and proteolytic activities. Furthermore, the relationship between lethality of ECPs and mortality caused by bacterial challenge was examined. Correlation was demonstrated between the pathogenic properties and proteolytic activities of the ECPs. Cytolytic (GCAT) activity comparable with that of the typical reference strain used (NCMB 1102) was not detected in ECPs of any of the atypical strain tested. An extracellular metallo-caseinase, AsaP1, was linked with lethal toxicity and a strong pathogenic effect. Furuncular-like lesions were produced by ECPs containing AsaP1 activity. One strain produced a lethal toxin which was neither caseinolytic nor with GCAT comparable activity. The examined atypical strains form at least three distinct groups based on different virulence mechanisms and extracellular proteases. PMID- 9418020 TI - Biodegradation studies of polyaromatic hydrocarbons in aqueous media. AB - Sixteen bacterial strains isolated from an activated sludge and Mycobacterium ssp. PYR-1 were tested for their ability to degrade polyaromatic hydrocarbons (PAHs). The bacterial strains Pasteurella ssp. (B-2) and Mycobacterium ssp. PYR-1 (AM) showed a high biodegradation potential of three- and four-ring PAHs. Bacterial strain AM was able to degrade up to 80% of three and four-ring PAHs (phenanthrene, fluoranthene and pyrene) within the first month of incubation, while the bacterial strain B-2 achieved the same biodegradation in 2 months. The metabolic pathway of PAH degradation was studied using fluoranthene and the bacterial strain AM. Ninety per cent of fluoranthene was biodegraded within the first 9 d of incubation when applied as a single substrate. Retention factor values from thin-layer chromatography studies, gas chromatography with mass selective detection and tandem mass spectrometry identified 9-fluorenone-1 carboxylic acid as one of the stable metabolic products and from this a fluoranthene biodegradation pathway is proposed. PMID- 9418021 TI - Variation within Mycobacterium scrofulaceum. AB - The purpose of this study was to identify Mycobacterium scrofulaceum reliably and rapidly and investigate diversity within the species. Fifty-four cultures were identified as Myco. scrofulaceum by preliminary cultural and biochemical tests, thin-layer chromatography and double diffusion. These strains were examined by PCR based on the 65 kDa heat stress protein gene, followed by restriction enzyme analysis of the product with BstEII and HaeIII. This produced seven groups, most with fewer fragments than had been reported previously. The technique was a rapid and reliable method for studying variation within Myco. scrofulaceum but alone, was unable to discriminate between some of these variants and other genetically similar species. When PCR-RFLP results were combined with biochemical tests, the major groups appeared to relate to different disease situations and thus, may have some epidemiological value. PMID- 9418022 TI - Spoilage microflora in fresh chicken breast stored at 4 degrees C: influence of packaging methods. AB - Chicken breasts with skin were packaged either in air, under vacuum or in modified atmospheres of (i) 30% CO2/70% N2 and (ii) 70% CO2/30% N2. After 3, 7, 14 and 21 days of storage at 4 degrees C, the samples were evaluated for spoilage microbial growth, odour and overall aspect. As expected, pseudomonads grew well in air or under vacuum, but growth was suppressed in both types of modified atmosphere packaging (MAP). However, growth of lactobacilli, Enterobacteriaceae and Brochothrix thermosphacta was not inhibited in MAPs. Modified atmosphere packaging (ii) extended shelf-life up to 21 days compared to 5 days for air packed samples. PMID- 9418023 TI - The prevalence of campylobacters and arcobacters in broiler chickens. AB - Chicken carcasses from a supermarket and from a poultry abattoir were examined using methods designed to isolate as many strains of campylobacters and related organisms as possible. Strains of arcobacter, but no campylobacters, were isolated from every carcass after enrichment. Campylobacter jejuni subsp. jejuni was isolated from all carcasses examined by direct plating and other Campylobacter-like strains were isolated from nine out of 15 abattoir carcasses by direct plating but not after enrichment. Only the Camp. jejuni subsp. jejuni strains could be identified to species level using a readily available identification scheme and/or a commercial identification kit. Examination of caecal contents from the 15 abattoir poultry yielded Camp. jejuni subsp. jejuni and Campylobacter-like strains from 15 and eight by direct plating, and from six and nine after enrichment, respectively. Four sites in the intestine of the abattoir birds (60 samples) were examined for arcobacters and only one strain was isolated. This indicates that arcobacters are probably not normal inhabitants of the poultry intestine. Poultry is a rich source of other campylobacteria besides the thermophilic Campylobacter spp. PMID- 9418024 TI - Bacteriophages of enteric bacteria in drinking water, comparison of their distribution in two countries. AB - The presence of bacteriophages infecting enteric bacteria was tested in more than 1500 drinking water samples in Israel and Spain. Bacteriophages tested were somatic coliphages, F-specific bacteriophages and Bacteroides fragilis bacteriophages. The three groups of bacteriophage were isolated in 100 ml water samples by the presence/absence test with similar frequencies, which ranged from 4.4% for somatic coliphages to 6.1% for bacteriophages infecting Bact. fragilis. In contrast, the frequency of isolation of bacteriophages was significantly higher than the frequency of isolation of faecal coliforms, which averaged only 1.9%. No significant differences were observed between the frequencies of isolation between the samples tested in Spain and those tested in Israel. The percentage of groundwater samples containing faecal coliforms and somatic coliphages was reduced significantly by chlorination, despite known deficiencies. However, there was no effect on the occurrence of F-specific bacteriophages and Bact. fragilis bacteriophages. PMID- 9418025 TI - Characterization of the rpsL and rrs genes of streptomycin-resistant clinical isolates of Mycobacterium tuberculosis in Japan. AB - Mutations in the rpsL and rrs genes associated with streptomycin resistance in Mycobacterium tuberculosis clinically isolated in Japan were characterized. The rpsL genes of 172 clinical isolates were amplified by PCR and classified into two groups on the basis of MboII restriction digestion. Thirty-three out of 54 (61.1%) streptomycin-highly resistant isolates (MIC > 200 micrograms ml-1) were not digested by MboII. By contrast, the remaining 21 of 54 (38.9%) streptomycin highly resistant isolates, all of 41 isolates with streptomycin resistance at a lower level (20 micrograms ml-1 < MIC < or = 200 micrograms ml-1), and all of 77 streptomycin-sensitive isolates, were restricted. Thus, all isolates resistant for MboII digestion showed a high level of resistance to streptomycin. Subsequently, the sequence for the rpsL and rrs genes from the 46 isolates were analysed. Eighteen out of 19 (94.7%) streptomycin-highly resistant isolates carried a mutation in any rpsL gene at position 43 or 88, or the rrs gene; 10 out of 17 (58.8%) streptomycin-resistant isolates at a lower level were confirmed to exhibit the mutation of either the mutated rpsL gene at position 88, or the rrs gene. In the total 36 streptomycin-resistant isolates, the mutation of the rpsL or rrs gene was observed in 28 streptomycin-resistant isolates, corresponding to 77.8%, whereas none of the streptomycin-sensitive isolates had mutations in either the rpsL or rrs gene. PMID- 9418026 TI - Detection of Campylobacter jejuni in food and poultry viscera using immunomagnetic separation and microtitre hybridization. AB - Thermophillic Campylobacter and Camp. jejuni were detected from samples of chicken liver, gall bladder, muscle and contaminated milk and chicken meat after an enrichment step by using immunomagnetic capture of cells with monoclonal antibody against a specific outer membrane protein of thermophilic Campylobacter. The detection of captured cells was achieved using two different hybridization methods. In one of the methods, the captured cells were lysed by guanidine isothiocyanate and the 23S rRNA was reacted with a microtitre plate-immobilized rDNA probe specific for thermophilic Campylobacter. In the other method, the captured cells were subjected to lysis by ultrasonication and the genomic DNA reacted with a microtitre plate-immobilized RNA probe specific for Camp.jejuni. Detection of the RNA-DNA hybrids formed in the wells was carried out using a monoclonal anti-RNA-DNA hybrid antibody. PMID- 9418027 TI - Antimicrobial activity among Pseudomonas and related strains of mineral water origin. AB - Siderophore production in 382 Pseudomonas and related strains of mineral water origin were screened and the antimicrobial activities of 158 of these tested against nine target organisms of health significance. Presence of siderophores could be detected in 54.4% and the majority of strains tested (91.2%) inhibited at least one of the nine target strains. Staphylococcus, Escherichia coli and Aeromonas hydrophila were particularly sensitive. Addition of iron eliminated the inhibitory activity in 96.7% of cases; the antagonistic effect should be largely determined by siderophore-mediated competition for iron. Most of the inhibitory strains produced siderophores, whereas the non-inhibitory strains did not. Few strains also produced bacteriocins showing activity against Pseudomonas aeruginosa and Aer. hydrophila. Strains isolated from mineral water have a broad antibacterial potential. PMID- 9418028 TI - Advisory Committee on Dangerous Pathogens (ACDP). Seminar on microbiological risk assessment, 28 January 1997. AB - Microbiological risk assessment is an area of growing importance and significant potential, where the underlying science, software systems and databases are developing to the point of real and useful application. It is also an area where the developing science is posing as many questions as it is presenting answers. Key issues emerging from the day included: the need for more sophisticated management of uncertainty, which is much more relevant to microbiological risk analysis than to other applications; the need for global surveillance systems with better compatibility and appropriate peer review; considered assessment of the impact of new molecular-based diagnostic and screening techniques; the explosion of relevant information available, particularly on the Internet, which makes computer literacy essential both to professionals and 'laymen'; and the appearance of software systems which are either tailored for microbiological application or have the potential for this use. The closely associated issues of risk communication and perception also emerged as being vital to the effective application of microbiological risk management to public health issues. Overall, the majority of participants considered the event to have been valuable and stimulating and thought that it would lead to improvements in the use of microbiological risk assessment. The Advisory Committee on Dangerous Pathogens is committed to taking this topic forward and will be both taking up the messages from this seminar and encouraging development of suitable databases and software systems. PMID- 9418029 TI - The mechanics of flight in the hawkmoth Manduca sexta. I. Kinematics of hovering and forward flight. AB - High-speed videography was used to record sequences of individual hawkmoths in free flight over a range of speeds from hovering to 5 ms-1. At each speed, three successive wingbeats were subjected to a detailed analysis of the body and wingtip kinematics and of the associated time course of wing rotation. Results are presented for one male and two female moths. The clearest kinematic trends accompanying increases in forward speed were an increase in stroke plane angle and a decrease in body angle. The latter may have resulted from a slight dorsal shift in the area swept by the wings as the supination position became less ventral with increasing speed. These trends were most pronounced between hovering and 3 ms-1, and the changes were gradual; there was no distinct gait change of the kind observed in some vertebrate fliers. The wing rotated as two functional sections: the hindwing and the portion of the forewing with which it is in contact, and the distal half of the forewing. The latter displayed greater fluctuation in the angle of rotation, especially at the lower speeds. As forward speed increased, the discrepancy between the rotation angles of the two halfstrokes, and of the two wing sections, became smaller. The downstroke wing torsion was set early in the halfstroke and then held constant during the translational phase. PMID- 9418030 TI - The mechanics of flight in the hawkmoth Manduca sexta. II. Aerodynamic consequences of kinematic and morphological variation. AB - Mean lift coefficients have been calculated for hawkmoth flight at a range of speeds in order to investigate the aerodynamic significance of the kinematic variation which accompanies changes in forward velocity. The coefficients exceed the maximum steady-state value of 0.71 at all except the very fastest speeds, peaking at 2.0 or greater between 1 and 2 ms-1. Unsteady high-lift mechanisms are therefore most important during hovering and slow forward flight. In combination with the wingtip paths relative to the surrounding air, the calculated mean lift coefficients illustrate how the relative contributions of the two halfstrokes to the force balance change with increasing forward speed. Angle of incidence data for fast forward flight suggest that the sense of the circulation is not reversed between the down- and upstrokes, indicating a flight mode qualitatively different from that proposed for lower-speed flight in the hawkmoth and other insects. The mid-downstroke angle of incidence is constant at 30-40 degrees across the speed range. The relationship between power requirements and flight speed is explored; above 5 ms-1, further increases in forward velocity are likely to be constrained by available mechanical power, although problems with thrust generation and flight stability may also be involved. Hawkmoth wing and body morphology, and the differences between males and females, are evaluated in aerodynamic terms. Steady state force measurements show that the hawkmoth body is amongst the most streamlined for any insect. PMID- 9418031 TI - Aerial performance of Drosophila melanogaster from populations selected for upwind flight ability. AB - A computerized system for three-dimensional tracking of large numbers of individual free-flying insects was used to assess the performance of Drosophila melanogaster from populations that had undergone 160 generations of selection for upwind flight ability. Compared with control lines, the selected lines showed significant increases in mean flight velocity, decreases in angular trajectory and a significant change in the interaction between velocity and angular trajectory. Maximal flight velocity was apparent as a sharply defined upper boundary of the distribution of horizontal and vertical velocity as a function of angular trajectory; this upper bound (0.85 ms-1) differed little between the selected and control lines, although individuals from the selected lines attained maximal performance levels much more frequently. Maximum induced power output was calculated directly from the product of maximum vertical velocity and body weight. This measure (28 W kg-1 muscle) was closely predicted by a scaling relationship derived from the load-lifting limits of larger insects and vertebrates, as well as tethered D. melanogaster stimulated via their optomotor reflex to produce maximal lift. These results indicate that selection for flight performance can readily alter the relative effort and/or the frequency of phenotypes capable of attaining population-wise maximal performance levels, but shows little ability to increase population-wise maximal performance. PMID- 9418032 TI - Flight and size constraints: hovering performance of large hummingbirds under maximal loading. AB - As the smallest birds, hummingbirds are the only birds capable of prolonged hovering. This suggests that hovering locomotion scales unfavourably with size. Is the hovering performance of larger hummingbird species more constrained by size than that of smaller ones? Maximal load-lifting capacities of the two largest species of hummingbirds found in the United States, the blue-throated (Lampornis clemenciae, 8.4 g) and magnificent (Eugenes fulgens, 7.4 g) hummingbird, as well as the two other local small species, the black-chinned (Archilochus alexandri, 3.0 g) and rufus (Selasphorus rufus, 3.3 g) hummingbird, were determined under conditions of short-burst performance. The power reserves of hummingbirds are substantial relative to normal hovering performance. The two large species lifted maximal loads close to twice their body mass for a very brief duration of over 0.4 s. The small species lifted maximal loads approximately equal to their own mass with a longer duration of over 0.6 s. For the two large species under maximal loading, estimates of burst muscle mass specific mechanical power output assuming perfect elastic energy storage averaged 309 W kg-1, compared with 75 W kg-1 during free hovering without loading. For the two small species, these values were 228 W kg-1 and 88 W kg-1, respectively. The differences in aerodynamic force production and power output between the large and small size classes occur despite their similar wing stroke velocity. This indicates that, during burst performance in these hummingbirds, the larger ones had a higher load-lifting capacity and generated more muscle power. In spite of the twofold difference in body mass, both large and small hummingbirds have evolved to become potent aerial competitors in order to exploit their common food resource, nectar. Both size classes have evolved to cope with the multi dimensional effects of size constraining their aerodynamics, muscle mechanics, metabolism and ecology. PMID- 9418033 TI - Seasonal variation in uptake of short-chain neutral amino acid by red blood cells and hepatocytes in trout (Salmo trutta). AB - The present study shows that the capacity of trout (Salmo trutta) red blood cells (RBCs) and freshly isolated hepatocytes to take up short-chain neutral amino acids changes according to a seasonal pattern. Maximal amino acid uptake rates in RBCs were obtained in winter and spring, while minima were seen in summer and autumn. In contrast, the maximal rates for the freshly isolated hepatocytes were obtained in autumn and winter, and the minima were seen in spring and summer. In addition, by studying the uptake of glycine, evidence was found that the activities of the amino acids carriers ASC, asc and Gly in RBCs varied according to a seasonal rhythm. The activity of the ASC and asc systems changed in parallel with the global uptake of amino acids. Moreover, the RBC:plasma concentration ratio for certain substrates of these carriers (alanine, serine and glycine) varied accordingly. In contrast, the activity of the Gly system was modified inversely with respect to the overall amino acid uptake. The activity of the ASC system in freshly isolated hepatocytes was also seasonally modified, reaching a maximum in autumn, shortly before the reproductive period. PMID- 9418034 TI - A comparison of two ELISA methods for the investigation of anti-cytomegalovirus IgG antibodies. AB - The Alpha-method ELISA for detection of anti-cytomegalovirus (CMV) IgG antibodies (test 1) was compared with another ELISA technique MEIA (test 2). Samples (248 sera and 56 cerebrospinal fluid) from patients with suspected CMV infection were investigated. Discordant samples were re-analysed, undiluted, with a latex test. Positive results were considered to be true positives when there was agreement in the results from the two methods. There were fourteen discrepant samples (4.6%). The latex agglutination test confirmed eight positive for test 1 and six positive for test 2. The overall diagnostic yield of tests 1 and 2 was sensitivity, 91% and 100%; specificity, 99% and 96%; positive predictive values, 95% and 85%; and negative predictive values 98% and 100%, respectively. PMID- 9418035 TI - Influence of gamma rays and sodium chloride on aflatoxin production by Aspergillus flavus. AB - The rate of Aspergillus flavus growth and production of aflatoxin in yeast extract sucrose medium generally decreased as the concentrations of NaCl increased from 2 to 12%. Maximum production and accumulation of aflatoxin at 28 degrees C occurred after 7 days in the presence of NaCl concentrations. The number of colony forming units of A. flavus after 1.0 or 2.0 kGy irradiation was lower than in the unirradiated controls and the mould was eliminated at 3.0 kGy. Aflatoxin B1 decreased from the control level of 46 micrograms kg(-1) to 10 micrograms kg(-1) at an irradiation dose level of 2.0 kGy. Viable gamma irradiated conidia (2.0 kGy) of A. flavus showed increased sensitivity to NaCl concentrations, indicating gamma-ray injury. The levels of aflatoxin produced by 2.0 kGy irradiated conidia of A. flavus decreased in the presence of 2, 4 or 6% NaCl, and the detoxification rate was 94.1, 100 and 100%, respectively, after 21 days of incubation at 28 degrees C compared with the effect of gamma-rays or sodium chloride alone. PMID- 9418036 TI - A new simple assay for determining aminoglycoside inactivation in intact cells of Pseudomonas aeruginosa. AB - Intact cells of aminoglycoside (AG) antibiotic-resistant Pseudomonas aeruginosa usually do not inactivate AG, even though they possess the AG-modifying enzyme. An assay method for determining the activity of inactivating enzyme in intact cells of streptomycin-resistant P. aeruginosa was previously reported. Although this assay method was applied to the determination of the activity of kanamycin (KM)-inactivating enzymes, it could not apply to some of the KM-resistant strains. A new simple assay method has now been investigated for determining the activity of KM-inactivating enzyme in intact cells of clinically isolated KM resistant P. aeruginosa. The determination of AG-inactivating enzyme activity was attempted using lysozyme for release of the inactivating enzymes in washed cells, and both DNase and RNase were added to digestion of the nucleic acids released by bacteriolysis. This lysozyme-DNase-RNase (LDR) method has facilitated the confirmation of the presence of AG-inactivating enzyme in the strains used. In addition, the LDR technique was applicable to the determination of inactivating enzyme activity for various AGs other than KM. Since this simple assay method can determine any type of AG-inactivating enzyme activity of various P. aeruginosa strains, it may contribute significantly to the rapid selection of drugs in clinical use. PMID- 9418037 TI - Some properties of gamma-glutamyl transpeptidase from Fusobacterium nucleatum. AB - A series of chromatographic techniques was used to purify gamma-glutamyl transpeptidase from Fusobacterium nucleatum ATCC 23726. The purified enzyme was homogenous with a molecular weight (MW) of approximately 101 kD consisting of two different subunits with MW of 67 kD and 31 kD. Its distribution by treatment of lysozyme-EDTA suggested that the enzyme was a periplasmic protein. The pl of the enzyme was 5.9 to 6.2, and the optimum pH of the transpeptidation and the hydrolytic reaction was 8.0. Glycylglycine, glycine and methionine were good acceptors, and the enzyme reaction, in the presence of glycylglycine, was especially efficient. Glutamic acid, glutamine and serine were poor acceptors, and the enzyme activity was inhibited by these amino acids. The apparent Km value for the gamma-glutamyl donor (gamma-glutamyl-p-nitroanilide) was 4.9 x 10(-4)M and that for the acceptor (glycylglycine) was 0.19 M. Affinity-labelling reagents, such as DON, azaserine and AT-125 strongly inhibited the enzyme activity, and its activity was inhibited by L-serine plus borate, as is mammalian gamma-glutamyl transpeptidase. The antibodies against gamma-glutamyl transpeptidase from bovine kidney reduced the activity of the bacterial enzyme by 65%. These results indicate that the catalytic sites in F. nucleatum gamma glutamyl transpeptidase were similar to those in mammalian gamma-glutamyl transpeptidase. PMID- 9418038 TI - Production of Escherichia coli group I-like heat-labile enterotoxin by Enterobacteriaceae isolated from environmental water. AB - Strains of Klebsiella oxytoca (2), Citrobacter freundii (2), Enterobacter cloacae (1) and Escherichia coli (1) isolated from environmental water were identified as heat-labile enterotoxin (LT) producing strains by immunological methods and polymerase chain amplification. A 322 bp amplified fragment was obtained with specific primers LTR and LTL, and hybridized to a digoxigenin-labelled LTB probe only under low stringency conditions, and not with a cholera toxin probe. These results suggest that Enterobacteriaceae may produce a LT-like toxin antigenically and genetically related to the LT enterotoxin of E. coli. PMID- 9418039 TI - Structural analysis of elements contributing to 5' splice site selection in plant pre-mRNA transcripts. AB - In vivo analyses of the cis-acting sequence requirements for pre-mRNA splicing in tobacco nuclei have previously demonstrated that 5' splice sites are selected by their position relative to AU-rich sequences within plant introns and by their degree of complementarity to the 5' end of U1 snRNA. To identify the specific nucleotide sequences promoting recognition of the 5' exon-intron boundary, multiple mutations have been introduced into a 22 nt AU-rich block positioned between two competing 5' splice sites in a derivative of pea rbcS3A intron 1. Transient expression of these mutant transcripts has delineated a 9 nt element positioned 30-38 nt downstream from the 5' splice site whose sequence composition affects 5' splice site choice. Uridine substitutions within this element do not alter 5' splice selection patterns, and, in fact, enhance recognition of the upstream +1wt 5' splice site slightly. Guanosine substitutions in this region, specifically creating an AG-rich AAAGGAGAGGCAGA motif (mutations shown underlined), reduce recognition of the upstream +1wt 5' splice site and enhance recognition of the downstream +106E 5' splice site. Cytosine substitutions at homologous positions marginally reduce recognition of the upstream 5' splice site. Additional mutations in this 9 nt element suggest that the AAAGGAGAGGCAGA motif acts as an specific 5' exon-defining element promoting recognition of downstream 5' splice sites, while AU-rich motifs promote recognition of upstream 5' splice sites. Mutations in an adjacent AU-rich region attenuate the effects of mutations in this short element but are not in themselves sufficient to alter 5' splice site selection. It is concluded that specific elements on both sides of the exon-intron boundary define the 5' splice site in this plant transcript. PMID- 9418040 TI - Mg-chelatase of tobacco: identification of a Chl D cDNA sequence encoding a third subunit, analysis of the interaction of the three subunits with the yeast two hybrid system, and reconstitution of the enzyme activity by co-expression of recombinant CHL D, CHL H and CHL I. AB - Mg-protoporphyrin IX chelatase catalyzes insertion of the magnesium ion into protoporphyrin IX, the last common intermediate precursor in chlorophyll and heme biosynthesis, to form Mg-protoporphyrin IX. In Rhodobacter sphaeroides, and Synechocystis, the three open reading frames bchD/chID, bchH/chIH and bchI/chII encode proteins which are required for in vitro Mg-chelatase activity. In higher plants also, three proteins are necessary for the Mg chelation, and genes homologous to bchH and bchI have been isolated previously. In this study, a novel tobacco cDNA sequence homologous to bchD is isolated and initially characterized. Together with the tobacco clones encoding the other two subunits, full-length cDNAs are now available for the first time for all three subunits of one plant species. The CHL D polypeptide deduced from the open reading frame encodes a protein of 758 aa (82.9 kDa) with an amino terminal extension that resembles a plastid transit peptide. Sequence comparison of tobacco CHL D revealed similarities to the D subunit of Rhodobacter and Synechocystis of 44% and 75%. The amino terminal half of CHL D shows significant similarity (46%) to the entire CHL I peptide sequence, indicating a gene duplication from an ancestral gene. The carboxy terminal half seemed to be unique. Both parts of CHL D are linked with a glutamine/asparagine/proline-rich region flanked by a highly acid-rich segment. Protein-protein interaction among the three subunits CHL D, H and I was studied using the yeast two-hybrid system. Physical interaction was demonstrated between CHL D and CHL I indicating that CHL D is part of the Mg-chelatase. Heterodimer formation of CHL H with CHL I or CHL D could not be demonstrated by transactivation of the lacZ reporter gene. Homodimerization of the CHL D subunit was indicated in the more sensitive assay on X-Gal-containing agar plates. In vitro Mg2+ insertion into protoporphyrin IX was demonstrated in protein extracts of yeast strains expressing the three subunits of tobacco Mg-chelatase. The reconstitution of the recombinant enzyme activity required additional ATP. PMID- 9418041 TI - A deletion mutation at the ep locus causes low seed coat peroxidase activity in soybean. AB - The Ep locus severely affects the amount of peroxidase enzyme in soybean seed coats. Plants containing the dominant Ep allele accumulate large amounts of peroxidase in the hourglass cells of the sub-epidermis. Homozygous recessive epep genotypes do not accumulate peroxidase in the hourglass cells and are much reduced in total seed coat peroxidase activity. To isolate the gene encoding the seed coat peroxidase and to determine whether it corresponds to the Ep locus, a cDNA library was constructed from developing seed coats and an abundant 1.3 kb peroxidase transcript was cloned. The corresponding structural gene was also isolated from a genomic library. Sequence analysis shows that the seed coat peroxidase is translated as a 352 amino acid precursor protein of 38 kDa. Processing of a putative 26 amino acid signal sequence results in a mature protein of 326 residues with a calculated mass of 35 kDa and a pl of 4.4. Using probes derived from the cDNA, genomic DNA blot hybridization and polymerase chain reaction analysis detected polymorphisms that distinguished EpEp and epep genotypes. Co-segregation of the polymorphisms in an F2 population from a cross of EpEp and epep plants shows that the Ep locus encodes the seed coat peroxidase protein. Comparison of Ep and ep alleles indicates that the recessive gene lacks 87 bp of sequence encompassing the translation start codon. Analysis by RNA blot hybridization shows that epep plants have drastically reduced amounts of peroxidase transcript compared with EpEp plants. The peroxidase mRNA is abundant in seed coat tissues of EpEp plants during the late stages of seed maturation, and could also be detected in root tissues, but not in the flower, embryo, pod or leaf. The results indicate that the lack of peroxidase accumulation in seed coats of homozygous recessive epep plants is due to a mutation of the structural gene that reduces transcript abundance. PMID- 9418042 TI - Specific interactions between the K domains of AG and AGLs, members of the MADS domain family of DNA binding proteins. AB - MADS domain (for MCM1, AG, DEFA and SRF) proteins are regulatory proteins found in all major eukaryotic kingdoms. Plant MADS domain regulatory proteins have a region of moderate sequence similarity that has been designated as the K domain, and its predicted coiled-coil structure suggests a role in establishing a protein protein interaction. In vivo studies with the Arabidopsis AGAMOUS (AG) protein have indicated that the K domain is important for AG function. Using a bait fusion protein containing the K domain and the C-terminal region of AG in a yeast two-hybrid selection, 156 clones that encode potential AG-interacting proteins were identified. These clones each encode one of four highly related MADS domain proteins: AGL2, AGL4, AGL6 and AGL9. Additional analysis showed that the K domain of AG alone was able to bind the K domains of these AGLs. This binding was further confirmed by immunoprecipitation experiments using in vitro synthesized AG and AGL K domains. These results strongly suggest that AG interacts with AGL2, AGL4, AGL6 and AGL9 in vivo. Based on these results and previous observations, it is proposed that the AG function requires interaction with at least one of these AGL proteins, and such interactions contribute to the functional specificity of the AG protein. PMID- 9418043 TI - An Arabidopsis mutant showing reduced feedback inhibition of photosynthesis. AB - Many plant genes are responsive to sugars but the mechanisms used by plants to sense sugars are unknown. A genetic approach has been used in Arabidopsis to identify genes involved in perception and transduction of sugar signals. For this purpose, an in vivo reporter system was established consisting of the light- and sugar-regulated plastocyanin promoter, fused to the luciferase coding sequence (PC-LUC construct). At the seedling stage, expression of the PC-LUC gene is repressed by sucrose, and a number of sucrose-uncoupled (sun) mutants were selected in which sucrose is unable to repress the activity of the PC promoter. Three mutants have been characterized in more detail. The sugar analog 2-deoxy-D glucose (2DG) was used to repress whole plant photosynthesis, PC-LUC gene expression and total ribulose-1,5-bisphosphate activity. It was found that the sun6 mutation makes plants unresponsive to these 2DG-induced effects. Moreover, unlike wild-type plants, sun6 mutants are insensitive to elevated levels of glucose in the growth medium. These findings suggest that the SUN6 gene is active in a hexose-activated signal transduction pathway. PMID- 9418044 TI - Light-induced transcriptional repression of the pea AS1 gene: identification of cis-elements and transfactors. AB - Here, we examine the cis-elements and trans-factors affecting the expression of asparagine synthetase (AS) genes whose transcription is negatively regulated by light. The promoters for the AS1 and AS2 genes of pea were isolated, sequenced, and functionally dissected for their ability to confer regulated expression to the GUS reporter gene in transgenic tobacco. Histochemical analysis of transgenic plants demonstrated that the AS1 and AS2 promoters show identical patterns of cell-specific expression. The more highly active AS1 promoter was further demonstrated to confer negative light-regulation to the GUS gene in transgenic tobacco. Deletion analysis and gain-of-function experiments showed that 124 bp of the AS1 promoter was sufficient to confer light-activated repression to a heterologous promoter. Potential conserved transcription regulatory elements, Box B, Box C, and Box C' within this region were shown to bind to nuclear proteins by gel shift analysis. A light-specific DNA:protein interaction was detected with Box B. The nuclear factors that bind to Box C and C' elements of AS1 are competed by a putative repressor element 'RE1' defined previously in the oat phytochrome gene whose transcription is also repressed by light. The Box B and C/C'-Box/RE1 binding factors were found in nuclear extracts of tobacco, pea, and Arabidopsis and may therefore be universal factors involved in light-activated transcriptional repression. PMID- 9418045 TI - The maize actin-depolymerizing factor, ZmADF3, redistributes to the growing tip of elongating root hairs and can be induced to translocate into the nucleus with actin. AB - The maize actin depolymerizing factor, ZmADF3, binds G- and F-actin, and increases in vitro actin dynamics. Polyclonal antibodies have been raised against ZmADF3 and these detect a single band of approximately 17 kDa in all maize tissues examined, with the exception of pollen. In the development of root hairs, the distribution of ZmADF3 is related to actin reorganization. In the early stages of hair development, ZmADF3 is distributed throughout the cytoplasm. As the hair emerges and the microfilament bundles redirect to the outgrowth there is a simultaneous redistribution of ZmADF3 which now concentrates at the tip of the emerging hair and remains in this position as elongation proceeds. These observations show that ZmADF3 localizes to a region where actin is being remodelled during tip growth. After cytochalasin D treatment which disrupts actin filaments, short rods of ZmADF3 and actin appear in the nucleus suggesting that ZmADF3 may function by guiding actin to sites of actin polymerization. PMID- 9418046 TI - Expression of a luteoviral movement protein in transgenic plants leads to carbohydrate accumulation and reduced photosynthetic capacity in source leaves. AB - Elucidating the role of viral genes in transgenic plants revealed that the movement protein (MP) from tobacco mosaic virus is responsible for altered carbohydrate allocation in tobacco and potato plants. To study whether this is a general feature of viral MPs, the movement protein MP17 of potato leafroll virus (PLRV), a phloem-restricted luteovirus, was constitutively expressed in tobacco plants. Transgenic lines were strongly reduced in height and developed bleached and sometimes even necrotic areas on their source leaves. Levels of soluble sugars and starch were significantly increased in source leaves. Yet, in leaf laminae the hexose-phosphate content was unaltered and ATP reduced to only a small extent, indicating that these leaves were able to maintain homeostatic conditions by compartmentalization of soluble sugars, probably in the vacuole. On the contrary, midribs contained lower levels of soluble sugars, ATP, hexose phosphates and UDP-glucose supporting the concept of limited uptake and catabolism of sucrose in the phloem. The accumulation of carbohydrates led to a decreased photosynthetic capacity and carboxylation efficiency of ribulose-1,5 bisphosphate carboxylase/oxygenase (rubisco) probably owing to decreased expression of photosynthetic proteins. In parallel, levels of pathogenesis related proteins were elevated which may be the reason for the obtained limited resistance against the unrelated potato virus Y (PVY)N in the transgenic tobacco plants. Ultrathin sections of affected leaves harvested from 2-week-old plants revealed plasmodesmal alterations in the phloem tissue while plasmodesmata between mesophyll cells were indistinguishable from wild-type. These data favour the phloem tissue to be the primary site of PLRV MP17 action in altering carbohydrate metabolism. PMID- 9418047 TI - Transgenic potato tubers accumulate high levels of 1-kestose and nystose: functional identification of a sucrose sucrose 1-fructosyltransferase of artichoke (Cynara scolymus) blossom discs. AB - By screening a cDNA library of artichoke (Cynara scolymus) blossom discs for fructosyltransferases, we isolated a clone designated Cy21. The deduced amino acid sequence shows homology to acid beta-fructosyl hydrolases and to the sucrose fructan 6-fructosyltransferase (6-SFT) of barley. Transiently expressed in Nicotiana tabacum protoplasts, the Cy21 gene-product synthesized 1-kestose, indicating that Cy21 codes for a sucrose sucrose 1-fructosyltransferase (1-SST). The enzyme worked at physiologically relevant sucrose concentrations (25 mM sucrose). In the protoplast system, 1-kestose seemed to be the only fructan product of the 1-SST. The enzyme activity was not affected by pyridoxal-HCl, an inhibitor of both the beta-fructosyl hydrolase and the fructosyltransferase activity of invertases. The fructosyltransferase activity of the Cy21 gene product, however, could be inhibited by Zn2+, Ag+ and Cu2+ ions. In artichoke plants the Cy21 transcript was highly abundant in primary roots and blossom discs. Transgenic potato tubers expressing Cy21 contain high levels of 1-kestose along with nystose and traces of fructosyl-nystose, supporting the conclusion that the Cy21 clone encodes a sucrose sucrose 1-fructosyltransferase. PMID- 9418048 TI - Calcium signalling in Arabidopsis thaliana responding to drought and salinity. AB - Changes in cytosolic free calcium concentration ([Ca2+]cyt) in response to mannitol (drought) and salt treatments were detected in vivo in intact whole Arabidopsis seedlings. Transient elevations of [Ca2+]cyt to around 1.5 microM were observed, and these were substantially inhibited by pretreatment with the calcium-channel blocker lanthanum and to a lesser extent, the calcium-chelator EGTA. The expression of three genes, p5cs, which encodes delta(1)-pyrroline-5 carboxylate synthetase (P5CS), the first enzyme of the proline biosynthesis pathway, rab18 and Iti78 which both encode proteins of unknown function, was induced by mannitol and salt treatments. The induction of all three genes by mannitol was inhibited by pretreatment with lanthanum. Salt-induced p5cs, but not rab18 and Iti78, expression was also inhibited by lanthanum. Induction of p5cs by mannitol was also inhibited by the calcium channel-blockers gadolinium and verapamil and the calcium chelator EGTA, further suggesting the involvement of calcium signalling in this response. Mannitol induced greater levels of p5cs gene expression than an isoosmolar concentration of salt, at both relatively high and low concentrations. However, calcium transients were of a similar magnitude and duration in response to both mannitol and isoosmolar concentrations of salt, suggesting that a factor other than calcium is involved in the discrimination between drought and salinity signals in Arabidopsis. In order to gauge the involvement of the vacuole as an intracellular calcium store in the response of Arabidopsis to mannitol, [Ca2+]cyt was measured at the microdomain adjacent to the vacuolar membrane. The results obtained were consistent with a significant calcium release from the vacuole contributing to the overall mannitol-induced [Ca2+]cyt response. Data obtained by using inhibitors of inositol signalling suggested that this release was occurring through IP3-dependent calcium channels. PMID- 9418049 TI - Expression of heterologous phytochromes A, B or C in transgenic tobacco plants alters vegetative development and flowering time. AB - In this study, oat phytochrome A (phyA), Arabidopsis phytochrome B (phyB) or Arabidopsis phytochrome C (phyC) were expressed in both day-neutral and photo period-sensitive (short-day) tobacco (Nicotiana tabacum cv. Hicks). Introgression of the Maryland Mammoth (MM) gene into cv Hicks was used to confer short-day photo-periodic sensitivity. Expression of oat phyA led to characteristic hypersensitivity of hypocotyls to red light (R) and far-red light (FR) and an overall dwarfing of the mature plant. Expression of Arabidopsis phyB enhanced the sensitivity of hypocotyls to R and caused even more marked dwarfing of the mature plant. In contrast, the expression of Arabidopsis phyC had no detectable consequences for the photocontrol of hypocotyl elongation. However, phyC expression did lead to a R-dependent increase in cotyledon expansion in de etiolating seedlings and to a significant increase in leaf area in mature plants. This provides the first experimental evidence that phyC is biologically active. The flowering time of cv Hicks plants grown under 8 h photoperiods was virtually unaffected by a 30 min white light (W) night break given 8 h into the dark period. In contrast, cv Hicks MM plants responded to a night break with a delay in flowering. Expression of phyA or phyB led to a night break-dependent delay in flowering in cv Hicks plants. For cv Hicks MM plants, the expression of any of phyA, phyB or phyC caused a marked enhancement of the flower-delaying effect of a night break. These observations indicate that transgenic phyA, phyB or phyC can interact with the endogenous mechanisms controlling flowering time in tobacco. PMID- 9418050 TI - Nicotiana plumbaginifolia hlg mutants have a mutation in a PHYB-type phytochrome gene: they have elongated hypocotyls in red light, but are not elongated as adult plants. AB - Two new allelic mutants of Nicotiana plumbaginifolia have been isolated which display a hypocotyl which is long (hlg) when seedlings are grown in continuous white light (W). This can be accounted for by the decreased response to red light (R) of the hypocotyl elongation rate in these mutants. Responses to other wavelengths are unaffected in the mutants. When grown in white light, mature hlg mutants are not elongated with respect to the wild-type; they also bolt and flower later. The shade-avoidance responses to red/far red ratio (R:FR) are intact in these mutants. Both mutants are deficient in phyB-like polypeptide that is immunodetectable in the wild-type; both have wild-type levels of a phyA-like polypeptide. These alleles are inherited in a partially dominant manner, and correspond to single-base missense mutations in a gene highly homologous to N. tabacum PHYB, which codes for a phytochrome B-type photoreceptor. One allele, hlg 1, has an introduced amino acid substitution; this may define a residue essential for phytochrome protein stability. The other allele, hlg-2, has a stop codon introduced C-terminal to the chromophore binding domain. As these phyB mutants are unaffected in shade-avoidance responses, but deficient in perception of R, it is concluded that the phyB absent in these mutants is responsible for R perception in the N. plumbaginifolia seedling, but is not a R:FR sensor in light grown plants. PMID- 9418051 TI - Two TIP-like genes encoding aquaporins are expressed in sunflower guard cells. AB - SunTIP7 and SunTIP20 are closely related sunflower cDNAs showing a deduced amino acid sequence homologous to proteins of the tonoplast intrinsic protein (TIP) family. Their expression in Xenopus oocytes caused a marked increase in osmotic water permeability (demonstrating that they are water channels) which was sensitive to mercury. In leaves, in situ hybridization revealed that both SunTIP7 and SunTIP20 mRNA accumulated in the guard cells. The possible involvement of SunTIPs in stomatal movement was examined by comparing the time course of transcript accumulation and leaf conductance during the daily cycle and following a water limitation. SunTIP7 mRNA fluctuations fitted changes occurring in leaf conductance. The transcript levels were markedly and systematically increased during stomatal closure. It is suggested that aquaporin SunTIP7 facilitates water exit associated with a decrease in guard cell volume. In the same conditions, the transcript level of SunTIP20 remained constant indicating that SunTIP genes are differentially regulated within the same cell. PMID- 9418052 TI - Compromising early salicylic acid accumulation delays the hypersensitive response and increases viral dispersal during lesion establishment in TMV-infected tobacco. AB - To investigate the role of salicylic acid (SA) in the hypersensitive response (HR) its accumulation was compromised during different phases of lesion development by differential expression of a salicylate hydroxylase gene (SH-L). Constitutive suppression of SA accumulation was achieved by expression of a gene fusion between the CaMV35S promoter (35S) and SH-L. Using the H2O2-responsive AoPR1 promoter to drive SH-L SA accumulation could be compromised at an early stage, on lesion formation and possibly prior to visible necrosis, whilst use of the salicylate-responsive PR1a promoter reduced SA accumulation at a later stage as lesions expand. TMV infection of 35S-SH-L and AoPR1-SH-L, but not PR1a-SH-L, tobacco resulted in significantly greater rates of lesion growth than in wild type tobacco. TMV was detected in asymptomatic tissue surrounding lesions only in 35S-SH-L and AoPR1-SH-L lines; subsequently these transgenic lines exhibited a 'spreading-necrosis' originating from the lesion which entered the stem and eventually other leaves, a phenotype which could be correlated with the presence of TMV particles. Analysis of TMV-infected and 'temperature-shifted' tobacco indicated that both 35S-SH-L and AoPR1-SH-L, but not PR1a-SH-L, transgenics exhibited delayed cell-death compared to wild-type infections. We propose that the SH-L phenotypes indicate that early SA accumulation is a major factor in preventing viral escape, via mechanism(s) which may include influencing the rate of host-cell death and, possibly, an effect on viral function. PMID- 9418053 TI - Interaction of Arabidopsis kinesin-like calmodulin-binding protein with tubulin subunits: modulation by Ca(2+)-calmodulin. AB - Kinesin-like calmodulin-binding protein (KCBP) is a recently identified novel kinesin-like protein that appears to be unique to and ubiquitous in plants. KCBP is distinct from all other known KLPs in having a calmodulin-binding domain adjacent to its motor domain. We have used different regions of KCBP to study its interaction with tubulin subunits and the regulation of this interaction by Ca(2+)-calmodulin. The results show that the carboxy-terminal part of the KCBP, with or without calmodulin-binding domain, binds to tubulin subunits and this binding is sensitive to nucleotides. In the presence of Ca(2+)-calmodulin the motor with calmodulin-binding domain does not bind to tubulin. This Ca(2+) calmodulin modulation is abolished in the presence of antibodies specific to the calmodulin-binding domain of KCBP. Similar binding studies with the carboxy terminal part of KCBP lacking the calmodulin-binding domain show no effect of Ca(2+)-calmodulin. These results indicate that Ca(2+)-calmodulin modulates the interaction of KCBP with tubulin subunits and this modulation is due to the calmodulin-binding domain in the KCBP. Calcium-dependent calmodulin modulation of KCBP interaction with tubulin suggests regulation of KCBP function by calcium, the first such regulation of a kinesin heavy chain among all the known kinesin like proteins. PMID- 9418055 TI - Promoter elements required for phloem-specific gene expression from the RTBV promoter in rice. AB - Previous studies indicated that a DNA fragment comprising nucleotides (nt) -164 to +45 of the RTBV promoter is sufficient to drive phloem-specific expression of a reporter gene in transgenic rice plants. In addition, two potential cis elements, Box I (nt -3 to +5) and Box II (nt -53 to -39) were identified by DNA protein interaction assays. In this study, the results of further in vivo studies involving mutagenesis of selected DNA sequences and analysis of expression of a reporter gene in transgenic rice plants revealed that, in addition to Box I and Box II, other elements are required for phloem-specific gene expression, among which are a direct repeat (ASL Box, nt -98 to -79) and a GATA motif (nt -143 to 135). All the these elements bind rice nuclear factors specifically, and mutations of the elements were identified that resulted in loss-of-competition in electrophoretic mobility shift assays. A DNA fragment comprising nt -164 to -32, which contains Box II, the ASL Box and the GATA motif, conferred phloem-specific reporter gene expression independent of Box I when fused to a heterologous CaMV 35S minimal promoter and introduced to transgenic rice plants. Studies that introduced point mutations suggested that in the context of the -103 to +45 promoter fragment, Box II and the ASL Box act synergistically to confer tissue specific gene expression. Similar studies in the -164 to +45 promoter fragment indicated that the -164 to -103 region, which includes the GATA motif, contains sequences that are functionally redundant with those in the -103 to -32 region, including the ASL Box and Box II. It is concluded that these regions act additively to direct phloem-specific gene expression. PMID- 9418054 TI - Overexpressed phytochrome C has similar photosensory specificity to phytochrome B but a distinctive capacity to enhance primary leaf expansion. AB - Phytochrome C (phyC) is a low-abundance member of the five-membered phytochrome family of photoreceptors in Arabidopsis. Towards developing an understanding of the photosensory and physiological functions of phyC, transgenic Arabidopsis plants were generated that over-express cDNA-encoded phyC and seedling responses to continuous white, red, or far-red light (Wc, Rc or FRc, respectively) were examined. Transgenic seedlings over-expressing phyC displayed enhanced inhibition of hypocotyl elongation in Rc, but were unchanged in responsiveness to FRc relative to wild-type. These data indicate that phyC has photosensory specificity that is similar to that of phyB and thus distinct from that of phyA. phyC overexpressors with levels only 3 to 4 times the level of endogenous phyC exhibited enhanced primary leaf expansion in Wc. This is in contrast to phyA or phyB overexpressors which respectively have levels that are 500- and 100-fold that of overexpressed phyC but showed no enhancement of primary leaf expansion. Therefore, phyC may have some physiological roles that are different to those of phyA and phyB in the control of seedling responses to light signals. PMID- 9418056 TI - Oxidative cross-linking of plasma membrane arabinogalactan proteins. AB - Monoclonal antibodies which recognize carbohydrate in arabinogalactan proteins (AGPs) have revealed that certain carbohydrate epitopes at the outer plasma membrane surface are developmentally regulated. Some epitopes are expressed according to cell position, and AGPs are thought to play a role in cell-cell interaction during development. This study demonstrates that sugar beet plasma membranes contain two subfamilies of AGPs, with apparent molecular masses of 82 and 97 kDa, and that each subfamily consists of a small number of acidic AGP isoforms. Excision of leaves generates three additional AGP complexes with apparent molecular masses of 120, 170 and 210 kDa, with the 170 kDa complex being the major form induced by excision. The addition of millimolar concentrations of H2O2 to a partially purified fraction of the 82 and 97 kDa AGPs also generates AGP complexes, with the 170 kDa complex as the major form. These results indicate that the plasma membrane AGPs are a target for endogenous H2O2. PMID- 9418057 TI - Map positions of 47 Arabidopsis sequences with sequence similarity to disease resistance genes. AB - Map positions have been determined for 42 non-redundant Arabidopsis expressed sequence tags (ESTs) showing similarity to disease resistance genes (R-ESTs), and for three Pto-like sequences that were amplified with degenerate primers. Employing a PCR-based strategy, yeast artificial chromosome (YAC) clones containing the EST sequences were identified. Since many YACs have been mapped, the locations of the R-ESTs could be inferred from the map positions of the YACs. R-EST clones that exhibited ambiguous map positions were mapped as either cleavable amplifiable polymorphic sequence (CAPS) or restriction fragment length polymorphism (RFLP) markers using F8 (Ler x Col-0) recombinant inbred (RI) lines. In all cases but two, the R-ESTs and Pto-like sequences mapped to single, unique locations. One R-EST and one Pto-like sequence each mapped to two locations. Thus, a total of 47 loci were identified in this study. Several R-ESTs occur in clusters suggesting that they may have arisen via gene duplication events. Interestingly, several R-ESTs map to regions containing genetically defined disease resistance genes. Thus, this collection of mapped R-ESTs may expedite the isolation of disease resistance genes. As the cDNA sequencing projects have identified an estimated 63% of Arabidopsis genes, a very large number of R-ESTs (approximately 95), and by inference disease resistance genes of the leucine-rich repeat-class probably occur in the Arabidopsis genome. PMID- 9418058 TI - AtLTP1 luciferase expression during carrot somatic embryogenesis. AB - The carrot (Daucus carota L.) EP2 gene encodes a Lipid Transfer Protein (LTP) which is expressed during protoderm formation in developing embryos. To develop a vital reporter system for gene expression during somatic embryo development a 1.1 kB fragment of the Arabidopsis thaliana LTP1 promoter was fused to the firefly luciferase (LUC) coding sequence. The AtLTP1 luciferase expression pattern in transformed carrot suspension cultures was identical to the expression pattern of the endogenous carrot EP2 gene. Cell tracking experiments revealed that all somatic embryos were derived from AtLTP1 luciferase expressing cell clusters. However, not all cell clusters that expressed the AtLTP1 luciferase reporter gene developed into a somatic embryo, suggesting that initiation of an embryogenic pathway in tissue culture does not always lead to development of a somatic embryo. PMID- 9418059 TI - Can synthetic pesticides be replaced with biologically-based alternatives?--an industry perspective. AB - Agricultural chemical companies have invested in the discovery and development of biological pesticides to complement synthetic pesticides for the control of insects, diseases, and weeds on agronomic and horticultural crops. For plant disease control, companies envisage biological fungicides entering markets where they have the best chance of performing and which are most receptive to using biological control methods. Fewer regulatory requirements can mean faster registration for a biological than a synthetic pesticide. However, industry's requirements for competitive performance, effective formulations, and economic production can mean significant investments in time and money for a biological pesticide, although total investment may be less than for a synthetic pesticide. One biocontrol project in which industry has invested is baculoviruses for insect control. Insect baculoviruses, genetically modified to kill insects faster than wild-type viruses, are attractive biocontrol agents because their selectivity to insect pests and safety to beneficial insects and mammals enable them to compete with synthetic insecticides. Industry is looking for similar biocontrol opportunities in disease control. Biocontrol agents for seedling disease, root rot, and postharvest disease control have been registered by the EPA and are trying to compete with synthetic fungicides for market share. To date, effective biocontrol agents have not been identified for the control of serious foliar diseases, such as grape downy mildew, potato late blight, wheat powdery mildew, and apple scab. Farmers must rely on synthetic fungicides and agronomic methods to control these diseases for the foreseeable future. PMID- 9418060 TI - Bacillus thuringiensis: from biodiversity to biotechnology. AB - Bacillus thuringiensis is a Gram-positive bacterium, widely used in agriculture as a biological pesticide. The biocidal activity mainly resides in a parasporal protein inclusion body, or crystal. The inclusion is composed of one or more types of delta-endotoxins (Cry and Cyt proteins). Cry proteins are selectively toxic to different species from several invertebrate phyla: arthropods (mainly insects), nematodes, flatworms and protozoa. The mode of action of the insecticidal proteins is still a matter of investigation; generally, the active toxin is supposed to bind specific membrane receptors on the insect midgut brush border epithelium, leading to intestinal cell lysis and subsequent insect death by starvation or septicemia. The toxin-encoding cry genes have been extensively studied and expressed in a large number of prokaryotic and eukaryotic organisms. The expression of such genes in transgenic plants has provided a powerful alternative for crop protection. PMID- 9418061 TI - Isolation and characterization of an antibiotic produced by the scab disease suppressive Streptomyces diastatochromogenes strain PonSSII. AB - An antibiotic produced by the scab disease-suppressive Streptomyces diastatochromogenes strain PonSSII has been isolated and partially characterized. The antibiotic is produced throughout culture growth, with maximum amounts accumulating in the broth when the culture is in the early stationary phase of growth. The activity declines within about 30 h after the culture enters stationary phase. Purification techniques included chromatography on Amberlite XAD-2, DEAE Sephadex and SP Sephadex in addition to C18 HPLC with an average yield of 75%. This antibiotic only inhibits pathogenic strains of S. scabies that cause scab disease on potato and other tuberous vegetables and does not affect S. griseus, S. venezuelae, Actinomyces bovis, Nocardia asteroides, Clostridium perfringens, Bacillus subtilis, Staphylococcus aureus, S. epidermidis, Enterococcus faecalis, Micrococcus luteus, Serratia marcescens and Escherichia coli. The antibiotic has a molecular weight of 500 or less, and is stable for weeks at acidic pH but is very labile at alkaline pH conditions. PMID- 9418062 TI - Evaluation of three decarboxylating agar media to detect histamine and tyramine producing bacteria in ripened sausages. AB - Histidine- and tyrosine-decarboxylase activity of 175 strains of bacteria isolated from eight retail samples of Spanish ripened sausages was tested in three decarboxylating agars (Niven medium, Joosten and Northolt medium and modified decarboxylating agar of Maijala) and confirmed by an enzymic method (histamine) and thin-layer chromatography (tyramine). Enterobacteria and pseudomonads showed the highest percentage of positive responses to histamine and tyramine in the three decarboxylating agars, but only enterobacteria were subsequently confirmed as histamine-producing. Confirmed tyramine-producing strains were all identified as enterococci or lactic acid bacteria. The medium described by Joosten and Northolt was more sensitive and faster at detecting tyramine-producing microorganisms. However, all three media failed to detect one histamine-positive strain of lactic acid bacteria used as a control. PMID- 9418063 TI - Endoglucanase production by paper-degrading mycoflora. AB - Fourteen fungal species, namely Aspergillus flavus, A. fumigatus, A. niger, A. ustus, Penicillium islandicum, P. wortmannii, Memnoniella echinata, Cladosporium herbarum, Stachybotrys atra, Chaetomium globosum, Fusarium oxysporum, Torula herbarum, Alternaria alternata and Curvularia uncinata were isolated from different grades of paper. They differ in their distribution on various kinds of paper and also in relative occurrence. While seasonal influence on mycoflora was observed, most of the moulds were capable of growing in all three seasons examined (summer, winter, rainy season). The moulds were cellulolytic in nature and endoglucanase activity was greatest in Aspergillus flavus, A. niger, A. fumigatus, P. wortmannii and P. islandicum. PMID- 9418064 TI - A simple method for evaluating Cryptosporidium-specific antibodies used in monitoring environmental water samples. AB - A simple method is described for the evaluation and quality control of Cryptosporidium-specific antibodies used in monitoring environmental water samples. Purified oocysts were fluorescently labelled with a test antibody at the appropriate concentration. Labelled oocysts were analysed using flow cytometry and a region was defined on a bivariate dotplot of fluorescence versus light scatter that enclosed all oocysts. Concentrates of environmental water samples that did not contain oocysts were then incubated with the test antibody and analysed using flow cytometry. The number of particles that appeared in the region defined for oocysts was recorded and was a measure of non-specific binding. The technique provides a simple, rapid and quantitative tool for both evaluating the binding specificity of test antibodies and optimizing sample staining conditions. PMID- 9418066 TI - Gene amplification (PCR) to detect and differentiate mycoplasmas in porcine mycoplasmal pneumonia. AB - The causative agent of porcine mycoplasmal pneumonia, Mycoplasma hyopneumoniae, is difficult and time-consuming to isolate. Serological identification using antibodies induced by the disease is confused by cross-reaction with a closely related organism, Myc. flocculare. From pig lungs obtained at slaughter for meat and deemed free of acute disease, it was possible to detect by culture both Myc. hyopneumoniae and Myc. flocculare. This study has improved on an earlier PCR detection of DNA from the former species by using a nested PCR capable of detecting the purified DNA equivalent to one mycoplasmal genome. With this PCR assay both mycoplasma species were detected and differentiated directly from lung tissue. PMID- 9418065 TI - Improved detection of enteroaggregative Escherichia coli using formalin-fixed HEp 2 cells. AB - Certain strains of enteroaggregative Escherichia coli cause detachment of HEp-2 cells during adhesion tests, preventing observation of the aggregative adherent phenotype. The use of formalin-fixed HEp-2 cells prevents cell detachment and facilitates the detection of enteroaggregative E. coli. PMID- 9418067 TI - Effects of sporulation pH on the heat resistance and the sporulation of Bacillus cereus. AB - Spores of Bacillus cereus ATCC 7004, 4342 and 9818 were obtained in nutrient agar at several pH from 5.9 to 8.3. The optimum pH for sporulation was around 7, but good production of spores was obtained in the range 6.5-8.3. With all three strains, D-values clearly dropped with sporulation pH, decreasing by about 65% per pH unit. z-Values were not significantly modified (P > 0.05) by this factor. Mean z-values of 7.13 degrees C +/- 0.16 for strain 7004, 7.67 degrees C +/- 0.04 for 4342 and 8.80 degrees C +/- 0.64 for 9818 were obtained. PMID- 9418068 TI - Susceptibility to vancomycin of enterococci isolated from dairy products. AB - The antibiotic vancomycin is often used in clinical therapy against multiple antibiotic-resistant bacteria. Twenty strains of enterococci, either Enterococcus faecium or Ent. faecalis, isolated from different cheeses were tested for resistance to vancomycin in liquid medium. Minimum inhibitory concentration (MIC) values ranged from less than 1 to 4 micrograms ml -1. A 399 bp intragenic fragment of the vanA gene was not observed after amplification of total DNA of the strains. It seems, therefore, that resistance to vancomycin, which is a common trait for enterococci of nosocomial origin, is not widespread among dairy isolates. PMID- 9418069 TI - PCR detection of Clostridium perfringens producing different toxins in faeces of goats. AB - A polymerase chain reaction (PCR) was used to identify the genes encoding the major toxins of Clostridium perfringens in faeces of goats. When pure cultures of Cl. perfringens types A, B, C, D and E were used as templates in the PCR, amplicons were observed on the agarose gel as bands at approximately the 247 (alpha primers), 1025 (beta primers), 403 (epsilon primers) and 298 (iota primers) bp level of the DNA marker. When used to identify different types of Cl. perfringens in samples artificially spiked with these micro-organisms, the PCR detected as few as 1-1.5 x 10(2) cfu g-1 of the five types of Cl. perfringens tested. The PCR technique allowed the identification and typing of Cl. perfringens strains in faeces of goats, without recourse to other techniques such as the mouse neutralization test. PMID- 9418070 TI - Cell-wall protein profiles of dairy thermophilic lactobacilli. AB - SDS-PAGE fingerprinting of cell-wall proteins extracted from 119 strains belonging to different species of lactic acid bacteria have been compared. The method of extraction and electrophoretic separation utilized in this work was found to be a reliable and rapid way for characterizing thermophilic lactobacilli species and strains. A protein of approximately 50 kDa was found to be characteristic for all the Lact. helveticus strains, and two cell-wall proteins of about 20 and 30 kDa were typical of the species Lact. delbrueckii, but the discrimination between the subspecies lactis and bulgaricus was not possible by the electrophoretic technique used. The other thermophilic species studied in this work presented cell-wall protein patterns that permitted their differentiation from both Lact. helveticus and Lact. delbrueckii. PMID- 9418071 TI - Ammonium ion affecting tylosin production by Streptomyces fradiae NRRL 2702 in continuous culture. AB - Nitrogen regulation in tylosin production by Streptomyces fradiae NRRL 2702 was studied in chemostat culture using a soluble synthetic medium. The maximum value of specific tylosin formation rate (qTYL) was 1.13 mg g-1 h-1 at the specific growth rate (mu) of 0.05 h-1, and qTYL decreased with increasing levels of the specific growth rate after reaching a rate of 0.1 h-1. The optimum conditions for tylosin formation were that the specific ammonium ion uptake rate (qN) and mu were 0.13 mmol g-1 h-1 and 0.05 h-1, respectively. The specific formation rates of threonine dehydratase (TDT) and tylosin were repressed by high levels of specific ammonium ion uptake rate. This study showed the adaptation to chemostat cultures of the nitrogen regulation of tylosin fermentations. PMID- 9418072 TI - Isolation and phenotypical identification of spore-forming Bacillus species from Jordanian habitats with larvicidal activity against Drosophila melanogaster. AB - Spore-forming Bacillus isolates were recovered from different Jordanian habitats. Of 37 samples, 187 colonies were selected. Forty-six (24.6%) of them were pathogenic to the third instar larvae of Drosophila melanogaster. Larvicidal activity of the isolates was from 0% (non-cultivated soil) to 83.3% (decomposed animal residues). The total spore count per gram weight varied from 0.1 x 10(5) 18 x 10(5) among the 37 tested samples. Morphological and microscopical identification of the isolates showed the presence of 16 different Bacillus species. The pathogenic isolates were B. thuringiensis (44) and B. sphaericus (2). PMID- 9418073 TI - Reappraisal of the effect of temperature on the growth kinetics of Aeromonas salmonicida. AB - The effect of temperature (1-34 degrees C) on the maximum specific growth rate of Aeromonas salmonicida could not be described by the classical growth models; for some strains, two optimal temperatures at 23 degrees C and 30 degrees C were observed, as well as an unexpected increase in the pseudolag time above 27 degrees C. This could be explained by the presence of two subsets, notably S layer+ and S-layer- sub-populations. The A- cells had higher growth parameters (Topt and mu opt) than the A+ cells and were selected by subcultures above 30 degrees C. Yet the relative proportion of A+ cells did not explain all the variation of mu max versus temperature, and the growth kinetics of an Aer. salmonicida isolate remained unpredictable. PMID- 9418074 TI - The use of ethidium bromide to assess a novel injury/recovery phenomenon in Listeria monocytogenes in inhibitory NaCl conditions. AB - An injury and recovery phenomenon was observed in Listeria monocytogenes inoculated into a medium containing 2.2 mmol l -1 NaCl, a concentration that was inhibitory to growth. The apparent loss then recovery of viability, as determined by plate counts, was compared with the uptake of ethidium bromide by the cells and found to be inversely related. Injury was caused not only by the initial osmotic up-shock but also by the subsequent down-shock involved in the spread plate protocol. PMID- 9418075 TI - Conversion of DL-threonine, D-threonine and 2-oxobutyrate into propionate and 2 hydroxybutyrate by Fusobacterium species. AB - The present investigation examined DL-threonine, D-threonine and 2-oxobutyrate conversion into propionate and 2-hydroxybutyrate by various type strains and clinical isolates of Fusobacterium. Except for Fus. naviforme, the type strains were able to produce varying degrees of propionate and/or 2-hydroxybutyrate from DL-threonine. Additionally, D-threonine was converted into an equimolar amount of propionate by Fus. necrophorum subsp. necrophorum, Fus. nucleatum subsp. nucleatum and Fus. varium, and to a lower but significant amount by Fus. mortiferum and Fus. perfoetens. However, the level of propionate remained unchanged for Fus. nucleatum subsp. fusiforme, Fus. nucleatum subsp. vincentii, Fus. naviforme, Fus. necrophorum subsp. funduliforme, Fus. gonidiaforme and Fus. russii. 2-Oxobutyrate was fermented to propionate by all type strains, although Fus. russii reduced it mainly to 2-hydroxybutyrate. Thus, an attempt was made to make use of these features in order to identify clinical isolates. PMID- 9418076 TI - Cryptosporidium parvum in environmental samples in the Sligo area, Republic of Ireland: a preliminary report. AB - To detect Cryptosporidium in environmental specimens in the Republic of Ireland, grab samples of river water were prepared by calcium carbonate flocculation, and marine mussel tissue homogenated prior to testing with a fluorescently labelled monoclonal antibody and fluorescence microscopy. The parasite was detected in both river waters and marine mussels (Mytilus edulis). Filter feeders such as Mytilus edulis may be of value as biological monitors for the presence of cryptosporidial oocysts in sea water. The presence of Cryptosporidium in river and marine waters and, in particular, contaminating mussels used for human consumption, has obvious health implications. PMID- 9418077 TI - Humanitarian action: the duty of all doctors. PMID- 9418078 TI - Human rights and medical education. PMID- 9418079 TI - Blinding laser weapons. PMID- 9418080 TI - South Africa: does a truth commission promote social reconciliation? PMID- 9418081 TI - Embargoes that endanger health. PMID- 9418082 TI - Prison health services. PMID- 9418083 TI - Strengthening "DOTS" through community care for tuberculosis. PMID- 9418084 TI - Canada re-examines its AIDS and HIV programmes. PMID- 9418085 TI - Death rate from HIV infection rises in Bombay. PMID- 9418086 TI - Systematic review of randomised controlled trials of strategies to promote adherence to tuberculosis treatment. AB - OBJECTIVE: To determine the effectiveness of strategies to promote adherence to treatment for tuberculosis. IDENTIFICATION: Searches in Medline (1966 to August 1996), the Cochrane trials register (up to October 1996), and LILACS (Literatura Latinoamericana y del Caribe en Ciencias de la Salud) (1982 to September 1996); screening of references in articles on compliance and adherence; contact with experts in research on tuberculosis and adherence. INCLUSION CRITERIA: Randomised or pseudorandomised controlled trials of interventions to promote adherence with curative or preventive treatment for tuberculosis, with at least one measure of adherence. MAIN OUTCOME MEASURE: Relative risks and 95% confidence intervals for estimates of effect for categorical outcomes. RESULTS: Five trials met the inclusion criteria. The relative risk for tested reminder cards sent to patients who defaulted on treatment was 1.2 (95% confidence interval 1.1 to 1.4), for help given to patients by lay health workers 1.4 (1.1 to 1.8), for monetary incentives offered to patients 1.6 (1.3 to 2.0), for health education 1.2 (1.1 to 1.4), for a combination of a patient incentive and health education 2.4 (1.5 to 3.7) or 1.1 (1.0 to 1.2), and for intensive supervision of staff in tuberculosis clinics 1.2 (1.1 to 1.3). There were no completed trials of directly observed treatment. All of the interventions tested improved adherence. On current evidence it is unclear whether health education by itself leads to better adherence to treatment. CONCLUSIONS: Reliable evidence is available to show some specific strategies improve adherence to tuberculosis treatment, and these should be adopted in health systems, depending on their appropriateness to practice circumstances. Further innovations require testing to help find specific approaches that will be useful in low income countries. Randomised controlled trials evaluating the independent effects of directly observed treatment are awaited. PMID- 9418087 TI - Comparison of cost effectiveness of directly observed treatment (DOT) and conventionally delivered treatment for tuberculosis: experience from rural South Africa. AB - OBJECTIVE: To conduct an economic evaluation of directly observed treatment (DOT) and conventionally delivered treatment for the management of new cases of tuberculosis in adults. DESIGN: Community based directly observed treatment, which has been implemented in the Hlabisa district of South Africa since 1991, was compared with a conventional approach to tuberculosis treatment widely used in Africa. Each was assessed in terms of cost, cost effectiveness, and feasibility of implementation within existing resource constraints. SETTING: Hlabisa Health District, South Africa. SUBJECTS: Adult patients with new cases of tuberculosis on smear testing; the number of cases increased from 20 per month to over 100 from 1991 to 1996. MAIN OUTCOME MEASURES: Cost of case management in 1996, cost effectiveness in terms of the cost per case cured, and bed requirements in comparison with bed availability for the 1990, 1993, and 1996 caseload. Costs are expressed in US dollars at values for 1996. RESULTS: Directly observed treatment was 2.8 times cheaper overall than conventional treatment ($740.90 compared with $2047.70) to deliver. Directly observed treatment worked out 2.4-4.2 times more cost effective, costing $890.50 per patient cured compared with either $2095.60 (best case) or $3700.40 (worst case) for conventional treatment. The 1996 caseload of tuberculosis required 47 beds to be dedicated to tuberculosis to implement directly observed treatment, whereas conventionally delivered treatment would have required 160 beds; the current number of beds for tuberculosis treatment in Hlabisa is fixed at 56. CONCLUSIONS: Because of the reduced stay in hospital, directly observed treatment is cheaper, more cost effective, and more feasible than conventional treatment in managing tuberculosis in Hlabisa, given the existing hospital bed capacity and the escalating caseload due to the HIV/AIDS epidemic. Such results may hold elsewhere, and wherever conventional tuberculosis management is practised a switch to directly observed treatment will increase hospital capacity to cope with a growing caseload. PMID- 9418088 TI - Mefloquine to prevent malaria: a systematic review of trials. AB - OBJECTIVE: To evaluate the research evidence on the efficacy and tolerability of mefloquine chemoprophylaxis. SEARCH STRATEGY: Any potentially relevant trial from the Cochrane Infectious Disease Group's register of controlled trials; systematic searches of Medline, Embase, Lilacs and Science Citation Index; scanning citations; and consulting drug companies and key investigators. We considered studies in all languages. INCLUSION CRITERIA: Trials carried out in non-immune adult travellers, and in non-travelling volunteers, where an attempt had been made to conduct a randomised comparison of mefloquine against placebo or against alternative standard prophylaxis. RESULTS: 37 potentially eligible trials of mefloquine prophylaxis were identified, and 10 met the inclusion criteria. These 10 trials comprised a total of 2750 non-immune adult participants randomised to mefloquine or to a control. One placebo controlled trial examined malaria incidence directly and showed mefloquine to be highly effective in preventing malaria in an area of drug resistance. However, four placebo controlled trials showed that mefloquine was not well tolerated, and withdrawals were consistently higher in mefloquine treatment arms than in placebo arms (odds ratio 3.49 (95% confidence interval 1.42 to 8.56)). Five field trials compared mefloquine with other chemoprophylaxis. Mefloquine was no worse tolerated than other chemoprophylaxis, although there was possibly a trend towards higher withdrawals in mefloquine arms (odds ratio 1.33 (0.75 to 2.36)). CONCLUSION: One trial showed mefloquine to be effective in preventing malaria, but withdrawal rates, presumably from side effects, were high across most studies. This is likely to impair mefloquine's effectiveness in general travellers, and it may therefore not be useful for routine prophylaxis. Mefloquine may be useful in specific situations such as for groups travelling to regions with a high risk of chloroquine resistant malaria and only limited access to effective medical care. PMID- 9418089 TI - Weapons injuries during and after periods of conflict: retrospective analysis. AB - OBJECTIVE: To assess the relative frequency of weapon injuries during conflict and after periods of conflict in the absence of disarmament. DESIGN: Retrospective analysis of a database of war wounds. SETTING: Region with a protracted conflict between rival combatant groups and a subsequent transition to the uncontested military authority of a single power. SUBJECTS: 2332 people who received weapons injuries during the conflict or post-conflict periods and were admitted to hospital within 24 hours of injury. MAIN OUTCOME MEASURES: Percentage change in mean monthly admission rate by weapon type between conflict and post conflict periods; annual incidence of injury by weapon type during conflict and post-conflict periods; percentage change in annual incidence by weapon type between conflict and post-conflict periods. RESULTS: Mean monthly admission rates for injuries from fragmentation munitions decreased by 8% between conflict and post-conflict periods and by 23% for injuries from mines and 32% for gunshot injuries. The decline in admissions for all injuries was 23%. After adjustment for population growth over the study period, declines in annual incidence were 22% for fragmentation munitions injuries, 34% for mine injuries, and 40% for gunshot injuries. The decline in incidence for all injuries was 33%. In-hospital mortality from weapons related injuries increased from 2.5% to 6.1% (P < 0.001) between conflict and post-conflict periods. CONCLUSIONS: In this setting, continued availability of weapons is associated with increased mortality and a level of injuries from weapons that is only somewhat reduced from that observed during a period of conflict. PMID- 9418090 TI - The quality of health care in prison: results of a year's programme of semistructured inspections. AB - OBJECTIVES: To assess, as part of wider inspections by HM Inspectorate of Prisons, the extent and quality of health care in prisons in England and Wales. DESIGN: Inspections based on a set of "expectations" derived mainly from existing healthcare quality standards published by the prison service and existing ethical guidelines; questionnaire survey of prisoners. SUBJECTS: 19 prisons in England and Wales, 1996-7. MAIN OUTCOME MEASURES: Appraisals of needs assessment and the commissioning and delivery of health care against the inspectorate's expectations. RESULTS: The quality of health care varied greatly. A few prisons provided health care broadly equivalent to NHS care, but in many the health care was of low quality, some doctors were not adequately trained to do the work they faced, and some care failed to meet proper ethical standards. Little professional support was available to healthcare staff. CONCLUSIONS: The current policy for improving health care in prisons is not likely to achieve its objectives and is potentially wasteful. The prison service needs to recognise that expertise in the commissioning and delivery of health care is overwhelming based in the NHS. The current review of the provision of health care in prisons offers an opportunity to ensure that prisoners are not excluded from high quality health care. PMID- 9418091 TI - Household survey of locomotor disability caused by poliomyelitis and landmines in Afghanistan. PMID- 9418092 TI - The validity of general practitioners' self assessment of knowledge: cross sectional study. AB - OBJECTIVE: To determine whether general practitioners can make accurate self assessments of their knowledge in specific areas. DESIGN: 67 general practitioners completed a self assessment of their level of knowledge over a variety of topics using a nine point semantic differential scale. An objective assessment of their knowledge was then made by administering true-false tests on two of the topics: thyroid disorders and non-insulin dependent diabetes. The study was repeated with another group of 60 general practitioners, using sexually transmitted diseases as the topic. SETTING: General practices in New Zealand. SUBJECTS: Random sample of 67 general practitioners in Auckland. MAIN OUTCOME MEASURE: Test scores for self assessment and for actual knowledge. RESULTS: Correlations between self assessments and test scores were poor for all three topics studied (r = 0.19 for thyroid disorders, 0.21 for non-insulin dependent diabetes, 0.19 for sexually transmitted diseases). CONCLUSIONS: As general practitioners cannot accurately assess their own level of knowledge on a given topic, professional development programmes that rely on the doctors' self perceptions to assess their needs are likely to be seriously flawed. PMID- 9418093 TI - General management of end stage renal disease. PMID- 9418094 TI - ABC of palliative care. HIV infection and AIDS. PMID- 9418095 TI - They were cheap and available: prisoners as research subjects in twentieth century America. PMID- 9418096 TI - To the point of farce: a Martian view of the Hardinian taboo--the silence that surrounds population control. PMID- 9418097 TI - Africa in the 21st century: can despair be turned to hope? PMID- 9418098 TI - Pitfalls of tuberculosis programmes in prisons. PMID- 9418099 TI - Abhorrent weapons and "superfluous injury or unnecessary suffering": from field surgery to law. PMID- 9418100 TI - Antipersonnel landmines: facts, fictions, and priorities. PMID- 9418101 TI - Walk in peace: banish landmines from our globe. PMID- 9418102 TI - Sudan: eating dust and returning to dust. PMID- 9418103 TI - Tajikistan: no pay, no care. PMID- 9418104 TI - Progress in reducing inpatient mortality from acute myocardial infarction is slow. PMID- 9418105 TI - Risk of testicular cancer in boys with cryptorchidism. Study was based on small number of cancers. PMID- 9418106 TI - Trial of thyroxine treatment for biochemically euthyroid patients has been approved. PMID- 9418107 TI - Doctors have moral imperative to call for end to embargo on Cuba. PMID- 9418108 TI - Reduction in use of temazepam is factor in deaths related to overdose. PMID- 9418109 TI - Local research ethics committees. Oxford committee was concerned that trial might be unethical. PMID- 9418110 TI - Local research ethics committees. Research discovers the right thing to do; audit ensures that it is done right. PMID- 9418111 TI - Local research ethics committees. Monitoring body is needed for audit. PMID- 9418112 TI - Total ban on landmines is unnecessary. PMID- 9418113 TI - Treating alcohol dependence. One glass of wine is usually 1.5 units. PMID- 9418114 TI - Treating alcohol dependence. Supplementation with parenteral B vitamins should be routinely considered. PMID- 9418115 TI - Treating alcohol dependence. Chlormethiazole is widely used in Europe. PMID- 9418116 TI - Lactic acidosis induced by phenformin is still a public health problem in Italy. PMID- 9418117 TI - Death of Diana, Princess of Wales. People experiencing emotional difficulties react in different ways. PMID- 9418118 TI - Death of Diana, Princess of Wales. Her death and funeral rate as traumatic stressors. PMID- 9418119 TI - Review of interventions to prevent heart disease. Study neglected to examine benefits of exercise. PMID- 9418120 TI - Academia: the view from below. Inner city scheme provides springboard for entry into academic general practice. PMID- 9418121 TI - Academia: the view from below. Academic medicine does not fit in with motherhood. PMID- 9418122 TI - Public health information on World Wide Web is hard to find. PMID- 9418123 TI - Persistence of functionally compromised anti-double-stranded DNA B cells in the periphery of non-autoimmune mice. AB - Both anti-single-stranded (ss) and anti-double-stranded (ds) DNA antibodies are associated with the autoimmune disease systemic lupus erythematosus (SLE), but only anti-dsDNA antibodies are considered one of the diagnostic criteria. Using Ig transgenes coding for anti-DNA we have determined the fate of anti-dsDNA B cells in a non-autoimmune environment. In a Rag-2 wild-type background, B cells expressing the anti-dsDNA Ig transgenes are present in the spleen but dsDNA specificity is disrupted due to expression of endogenous L chains. In a Rag-2 deficient background where co-expression of endogenous Ig is blocked, splenic B cells expressing only the anti-dsDNA transgene Ig are present, indicating that endogenous Ig expression is not required for bone marrow export. The anti-dsDNA B cells that persist are profoundly crippled in that they are unable to proliferate to lipopolysaccharide or anti-Ig stimulation. Furthermore, these anti-dsDNA Ig transgene B cells show a decreased lifespan relative to non-transgene BALB/c B cells. Persistence of anti-dsDNA B cells in the periphery of non-autoimmune mice raises the possibility that their appearance in the context of SLE is due to their reactivation by T cell help. PMID- 9418124 TI - Selective disruption of lymphotoxin ligands reveals a novel set of mucosal lymph nodes and unique effects on lymph node cellular organization. AB - Lymphotoxin (LT) provides a critical signal for the genesis of lymph nodes (LN) in mice. Here we show that mice treated in utero with LT beta-R-Ig, which binds to the membrane LT alpha 1 beta 2 heterotrimer, lacked most LN, yet retained a set of mucosal surface draining LN. Since mice genetically deficient in LT alpha lack all LN, including the mucosal set, we hypothesize that a novel LT alpha dependent pathway controls their genesis. This novel set of mucosal LN cannot be discriminated on the basis of addressin expression. The discovery of LN in mice treated with LT beta-R-Ig fusion protein in utero allowed us to compare the roles of membrane LT alpha beta or soluble LT alpha/tumor necrosis factor (TNF) in the development of cellular organization in LN and spleen. Our results indicate that both membrane LT alpha beta and soluble LT alpha/TNF mediate T-B cell segregation and the organization of B cell follicles in spleen and LN. Interestingly, while antagonism of membrane LT alpha beta or soluble LT alpha/TNF prevented germinal center (GC) formation in spleen, antagonism of soluble LT alpha/TNF had no effect on LN formation. The data suggest that multiple LT/TNF ligands control B cell follicle organization in the spleen and LN of adult mice, and that the requirements for LT/TNF ligands in GC formation are distinct in the different lymphoid organs. PMID- 9418125 TI - Induction of humoral and cellular immunity against influenza virus by immunization of newborn mice with a plasmid bearing a hemagglutinin gene. AB - Neonates and infants display an intrinsic disability to mount protective immune responses to influenza viruses or conventional influenza vaccines. We investigated the ability of naked DNA to prime protective immune responses by inoculating newborn and adult mice with a plasmid (pHA) expressing hemagglutinin (HA) from the neurovirulent strain A/WSN/33 of influenza virus. Continuous exposure to small doses of antigen subsequent to neonatal DNA immunization led to effective priming of specific B and Th cells, rather than tolerance induction. The pHA immunization of adult mice primed a strongly biased Th1 response, whereas in neonates it induced a mixed Th1/Th2 response. In contrast to the effect of live-virus immunization, DNA immunization of neonates was followed by enhanced cytotoxic T lymphocyte responses subsequent to challenge with A/WSN/33 influenza virus. Mice immunized as neonates or adults with pHA plasmid exhibited significant increases in survival and decreases in virus lung titers following lethal challenge with the A/WSN/33 virus or the A/PR8/34 drift variant. Our results demonstrate that DNA vaccination is an efficient and safe means to generate broad humoral and cellular immune responses to influenza viruses, during the earliest stages of postnatal life. PMID- 9418126 TI - Effect of the presence of transgenic H and L chain genes on B cell development and allelic exclusion. AB - The expression of various components of the antigen receptor is closely associated with the developmental progression of B lymphocytes. For this reason, introduction of rearranged Ig transgenes exerts profound effects on B cell development. In these studies we show that the presence of transgenes for both mu and delta H as well as L chains accelerates the rate of B cell maturation resulting in the appearance of large numbers of IgD-expressing B lymphocytes in vitro that are otherwise undetectable. In addition, allelic exclusion, normally exerted very effectively by these particular transgenes (carried in the MD-3 strain), was lost in the long-term bone marrow cultures. These findings can be recapitulated in vivo by serial adoptive transfers of bone marrow cells from the transgenic animals into lethally irradiated recipients. We conclude from these studies that allelic exclusion is not necessarily mediated by any one event but may be a result of the integrated occurrence of Ig H chain gene rearrangement, accumulation of transcription factors, along with the ordered progression of B cell differentiation under the influence of the appropriate inductive microenvironment. These findings may account for at least some of the observed cases of allelic inclusion in transgenic animals. PMID- 9418127 TI - B7 blockade prevents activation-induced cell death of thymocytes. AB - Although both B7 and its counter-receptor CD28 are expressed in the thymus, the role of B7 in thymic selection is not clear. We investigated the role of B7 in intrathymic deletion of antigen-specific T cells using a TCR transgenic model specific for antigen ovalbumin (OVA) and H-2Ad. Intraperitoneal injection of OVA induced apoptosis of thymocytes and drastic reduction of thymocyte numbers. This was significantly inhibited by co-injection of CTLA-4-Ig which blocks B7 co stimulation. Deletion of T cells in the thymus following i.p. injection of OVA was associated with T cell pre-activation as demonstrated by T cell proliferation and cytokine production. Injection of CTLA-4-Ig blocked all these activation events and rescued thymocytes from activation-induced cell death. These results demonstrate that B7 is required for the activation-induced cell death of MHC class II-restricted thymocytes in vivo. PMID- 9418128 TI - Specificity of CD8+ T cells from subunit-vaccinated and infected H-2b mice recognizing the 38 kDa antigen of Mycobacterium tuberculosis. AB - CD8+ T cells have been implicated in protective anti-tuberculous immune responses, but little is known about the identity of mycobacterial antigens recognized by CD8+ T cells. In this study we identified the Mycobacterium tuberculosis 38 kDa protein as a target for murine CD8+ cytotoxic T lymphocytes (CTL) which were induced by vaccination of C57BL/6 mice with DNA delivered with a plasmid, with transfected tumour cells or by infection with tubercle bacilli. Using overlapping synthetic peptides covering the whole protein sequence, peptides predicted to contain H-2Kb or H-2Db motifs, as well as naturally processed peptides, we were able to identify CTL epitopes. Differences were demonstrated in peptide specificity between CTL from immunized or M. tuberculosis infected mice. The identified CTL epitopes could be important for future analysis of the involvement of CD8+ T cells in M. tuberculosis infections and for vaccine development. PMID- 9418129 TI - B lymphocyte sensitivity to IgM receptor ligation is independent of maturation stage and locally determined by macrophage-derived IFN-beta. AB - Compartmentation of B lymphocyte populations is associated with differences in both development stage and sensitivity to Ig (sIg)-dependent triggering. In order to characterize the factors that contribute in setting the level of sensitivity of a B cell, we quantified sIgM-dependent regulation of Ig secretion in purified mature and immature B cells after ex vivo and in vivo modification of their environment. These analyses formally demonstrate that the bone marrow (BM) microenvironment locally induces high B cell sensitivity to sIgM ligation irrespective of differentiation stage. We further provide evidence that BM macrophages create a dominant environment that enhances B cell sensitivity to B cell receptor triggering. Finally, using ex vivo assays as well as type I IFN receptor-deficient mice we show that IFN-beta produced by resident BM macrophages is necessary and sufficient to define B cell sensitivity. Implications of these findings for the understanding of B cell selection processes are discussed. PMID- 9418130 TI - T cell response of I-Aq mice to self type II collagen: meshing of the binding motif of the I-Aq molecule with repetitive sequences results in autoreactivity to multiple epitopes. AB - Type II collagen (CII) is of immunological interest because of its repetitive structure and properties as an autoantigen. The mouse gene has recently been cloned, thus enabling T cell-defined epitopes to be identified. Multiple novel epitopes on mouse CII are here detected in the autoreactive T cell response. The major response is directed to an epitope with residues 707-721 located on the CB10 fragment. Some 25 other epitopes are also recognized, including the autologous homologue of the 256-270 epitope which dominates in the response to foreign collagen. The cells reactive with mouse collagen peptides were of Th1 type, as judged by release of IFN-gamma. No significant reactivity was detected to mouse CII peptides during ongoing disease. Alignment of the mouse epitopes revealed a sequence motif with characteristic side chains at residues P1, P4 and P7, and to a lesser extent at P5, within a nonamer core sequence. Binding of these epitopes was simulated in a computer model of the I-Aq molecule, where peptides with anchor residues at P1, P4 and P7 were indeed found to fit the binding groove best. The spacing of pockets and the fine structure of the binding surface of the I-Aq molecule meshes with the repetitive structure of the collagen (X-Y-Gly), thus providing a likely explanation for the occurrence of multiple epitopes. Comparison with human DR binding motifs showed that the I-Aq motif resembles most closely that of the DR4 subtypes which predispose for rheumatoid arthritis. PMID- 9418131 TI - The critical role of a solvent-exposed residue of an MHC class I-restricted peptide in MHC-peptide binding. AB - The immunodominant ovalbumin257-264 (OVA-8, SIINFEKL) and herpes simplex virus gB496-503 (HSV-8, SSIEFARL) peptides share 50% amino acid identity (residues P1, P3, P5 and P8) and bind with comparable efficacy to the murine MHC-encoded class I molecule H-2Kb. However, these two peptides bind differently to H-2Kbm8, a natural H-2Kb variant with a substitution in four amino acids on the floor of the peptide-binding site; HSV-8 binds with high and OVA-8 with a relatively low efficacy. To investigate which of the non-homologous peptide residues were responsible for this differential binding, we used substituted peptide variants and the class I thermodynamic stabilization assay. Variation at the solvent exposed peptide residues P6 and P7 did not appreciably influence binding. By contrast, variation at the buried P2 and, surprisingly, at the solvent-exposed P4 residue was found to be important. Transplantation of the HSV-8 P2 or P4 residues onto the OVA-8 backbone created variant peptides O2S (P2I-->S) and O4E (P4N-->E) that bound considerably better to H-2Kbm8 than OVA-8. Furthermore, the double substituted peptide, O2S4E, bound even better, revealing a cooperative effect of the two residues. The reciprocally substituted peptides H2I and H4N, generated by grafting the OVA-8 P2 and P4 residues onto the HSV-8 backbone respectively, bound to H-2Kbm8 slightly worse than HSV-8 but the double-substituted peptide H2I4N bound as poorly as OVA-8. Effects exerted by the P4 residue, which is solvent accessible and therefore available for the TCR contact, demonstrated that exposed peptide residues can, in certain situations, influence not only the TCR contact but also MHC-peptide binding. PMID- 9418132 TI - Wortmannin, a phosphatidylinositol 3-kinase inhibitor, blocks the assembly of peptide-MHC class II complexes. AB - Peptide-class II complexes are assembled in endocytic, lysosome-like compartments where newly synthesized class II molecules are targeted from the trans-Golgi network (TGN). Recent studies have implicated phosphatidylinositol 3-kinase (PI3 kinase) as an essential component in membrane trafficking from the TGN to lysosomes. Here, using subcellular fractionation, we show PI3-kinase activity associated with subcellular fractions which contain the class II peptide-loading compartment (IIPLC) in B cells. At concentrations required for inhibition of PI3 kinase activity in vivo, wortmannin blocked the processing and presentation of antigen by B cells to T cells. Treatment of B cells with wortmannin significantly limited the proteolytic degradation of invariant chain and the formation of peptide-class II complexes. Subcellular fractionation coupled with pulse-chase analyses showed that invariant chain and class II molecules trafficked to the IIPLC in wortmannin-treated cells. However, wortmannin prevented the maturation and correct targeting to the IIPLC of cathepsin D, a protease necessary for the degradation of invariant chain and assembly of processed antigen-class II complexes. These results suggest that li-class II complexes traffic to the IIPLC via a pathway that is relatively insensitive to wortmannin, but suggest a role for PI3-kinases in the trafficking of other components necessary for the assembly of processed antigen class II complexes to the IIPLC. PMID- 9418133 TI - Mapping the minimal murine T cell and B cell epitopes within a peptide vaccine candidate from the conserved region of the M protein of group A streptococcus. AB - The highly conserved C-terminus of the M protein of group A streptococcus (GAS) is a promising vaccine candidate. An epitope within the conserved C-terminus of the M protein, peptide 145 (a 20-mer with the sequence: LRRDLDASREAKKQVEKALE), has been defined which is the target of opsonic antibodies in both humans and mice, and is recognized by the sera of most adults living in areas of high streptococcal exposure. However, due to potential cross-reactivity between T cells stimulated by this region of the M protein and host cardiac myosin, it is critical to define precisely the minimal protective epitopes within p145. Studies have shown that the immunodominant epitope expressed by p145 is conformational, occurring as an alpha-helical coiled-coil. To enable us to map the murine minimal B cell and T cell epitopes within p145, we have used a novel strategy that allowed us to present shorter sequences of p145 in a native-like conformation. The minimal B cell epitope was found to be contained within residues 7-20 of the p145 sequence, and we have shown that mice immunized with this region are able to generate antibodies that bind to and also opsonize the organism GAS. The T cell epitope is located at the N-terminal region of the p145 sequence, residues 3-14. We have managed, therefore, to define a vaccine candidate--a minimal opsonic B cell epitope within the p145 sequence--that does not incorporate a potentially deleterious T cell epitope. PMID- 9418134 TI - Purification, and biochemical and biological characterization of an immunosuppressive and lymphocyte mitogenic protein secreted by Streptococcus sobrinus. AB - An immunosuppressive/mitogenic (ISM) protein was purified from the supernatants of cultures of Streptococcus sobrinus with an isoelectric point of 4.75 and a relative molecular mass of 38 kDa (p38). Treatment of C57BL/6 mice with p38 induced an increase in the numbers of non-specific splenic Ig-secreting plaque forming cells (PFC) with peak responses on day 3 for IgM-secreting PFC and on day 5 for IgG-secreting PFC, with an isotype pattern consisting predominantly of IgG2a and IgG2b. This increase was accompanied by a lymphocyte blastogenic response of both T and B lymphocytes. The in vitro effects of p38 on pure B, T and total splenic lymphocytes indicated that this ISM protein was primarily a B cell mitogen, being T cells activated subsequently by the generation of B blasts. Suppression of the murine primary immune response against sheep red blood cells was observed in C57BL/6 mice treated 4 days before with p38. The amino acid sequence of the N-terminus of p38 has a significant similarity with several enolases, particularly with rabbit enolase. However, the biological effects ascribed to p38 have not been detected after in vivo treatment with that enolase. The immunosuppressive effect of p38 was abrogated by depletion of IL-10 but not of IL-4. In agreement with this observation IL-10 was the only cytokine detected in serum of C57BL/6 mice after p38 treatment and the peak of serum levels was observed as soon as 2 h after treatment. PMID- 9418135 TI - Selection of a diverse TCR repertoire in response to an Epstein-Barr virus encoded transactivator protein BZLF1 by CD8+ cytotoxic T lymphocytes during primary and persistent infection. AB - We investigated the CD8+ cytotoxic T lymphocyte (CTL) repertoire to an HLA B8 restricted peptide, RAKFKQLLQ, located in the Epstein-Barr virus (EBV) immediate early protein, BZLF1. Repertoire selection was monitored by determining the TCR beta chain sequences of RAKFKQLLQ-specific CTL established from primary infected and healthy virus carriers. PCR analysis of spontaneous EBV-transformed lymphoblastoid cell lines (LCL) from three individuals with primary infection showed that two were infected with type A and one with type B EBV. Polyclonal and clonal CTL that were generated by stimulating peripheral blood mononuclear cells with an HLA B8+ homozygous LCL lysed T cell blasts pulsed with the peptide, RAKFKQLLQ; lysis of certain HLA B8+ LCL targets was associated with the abundance of BZLF1 transcripts. TCR beta analysis showed that while there was loop length restriction in the putative peptide contact site of all responding beta chains, diverse and unique (non-recurrent) TCR beta clonotypes were selected in individuals during primary infection and continued to emerge after long-term virus exposure. TCR-contact site heterogeneity was excluded as the selective force in diversity generation since the epitope-encoded sequences were found to be identical within endogenous virus isolates. In this first study of TCR repertoire selection for an EBV lytic antigen, a BZLF1-reactive component of diverse clonotypes was identified in primary type A or type B EBV infection which was sustained in the EBV-specific memory response throughout life-long infection. This diversity selection is likely to play a critical role in maintaining a balanced viral load throughout EBV persistence. PMID- 9418136 TI - Cooperation of B cells and T cells is required for survival of mice infected with vesicular stomatitis virus. AB - To define the role of T cells and B cells in resistance to vesicular stomatitis virus (VSV) infection, knockout mice with different specific immune defects on an identical background were infected i.v. and the outcome of infection was compared; in this way a more complete picture of the relative importance of various host defence mechanisms could be obtained. Compared to T and B cell deficient SCID mice which all succumbed from encephalitis within 5-9 days of infection, T cell-deficient nude mice generally lived longer, but within a period of approximately 1 month after challenge all died. In contrast, B cell-deficient mice were highly susceptible even to low doses of virus and mortality could be prevented by transfer of naive B cells prior to challenge as well as by immune serum given after challenge. Analysis of MHC class I- and class II-deficient mice revealed that CD8+ T cells could exert some antiviral activity, but CD4+ T cells sufficed for survival and were required for optimal resistance. Consistent with this it was found that in nude mice a lethal outcome could be prevented by transfer of CD8-depleted cells from B cell-deficient mice. Thus our results clearly demonstrate that while antibodies are pivotal for survival in the early phase of VSV infection, T cells are required for long-term survival, with CD4+ T cells being more effective in controlling this infection than CD8+ T cells. PMID- 9418137 TI - CD79 alpha/CD79 beta heterodimers are expressed on pro-B cell surfaces without associated mu heavy chain. AB - During B cell development, the surface expression of CD79 alpha/CD79 beta heterodimers had been thought to begin in the pre-B cell stage where the heterodimers constitute pre-B cell receptors together with mu heavy and surrogate light chains. Thereafter, in mature B cells, CD79 alpha/CD79 beta associates with surface Ig to form B cell antigen receptors. In this study, we revealed by using newly established mAb that CD79 beta was expressed on the surface of pro-B cells which had not undergone the productive Ig gene rearrangement. Biochemical analysis showed that CD79 beta on pro-B cells existed either as monomers or as disulfide-linked heterodimers with CD79 alpha, non-covalently associated with four unidentified membrane molecules. Our finding that CD79 beta is expressed on earlier B-lineage cells than previously expected coincides with the recent study in which CD79 beta-deficient mice exhibit a blockade of B cell differentiation at the pro-B cell stage. Thus, it is speculated that the CD79 beta-containing complexes on pro-B cell surfaces may function to induce early B cell differentiation. PMID- 9418138 TI - Antibody class switch recombinase activity is B cell stage specific and functions stochastically in the absence of 'targeted accessibility' control. PMID- 9418139 TI - Condylar reconstruction in extensive ankylosis of temporomandibular joint in adults using resected segment as autograft. A new technique. AB - A new method of condylar reconstruction after gap-arthroplasty in extensive ankylosis of the temporomandibular joint in adults is presented. An autogenous graft, consisting of the excised ankylotic mass, was contoured, reimplanted and used for condylar reconstruction. This technique has given good results and is a simple alternative to other methods of reconstruction in adults. Successful results in three cases with more than 12 months' follow up, has prompted us to use this technique routinely in adult patients. PMID- 9418140 TI - Otoplasty: a combined approach to different structures of the auricle. AB - A technique for otoplasty is presented, which combines the advantages of different methods. The procedure includes a dorsal skin excision, a cartilage incision at the border between the concha and scapha, scoring of the crus superior on the anterior side, and if necessary a reduction of the conchal height and modification of the position of the cauda helicis by cartilage excision. The results of the treatment of 526 ears in 312 patients are retrospectively analysed by a patient questionnaire and a chart review. PMID- 9418142 TI - Primary synovial chondromatosis of the temporomandibular joint with suspected traumatic etiology. Report of a case. AB - Synovial chondromatosis (SC) of the temporomandibular joint (TMJ) is a rare disease that is characterized by the development of nodules of cartilage within the synovial connective tissues of articulating joints. Reports of extracapsular TMJ SC are rare. A case is presented of primary SC of the TMJ with extension to the pterygoid plates, with a suspected traumatic etiology. The differences between primary and secondary SC are discussed. PMID- 9418141 TI - Single-channel thin-fiber and Nd:YAG laser temporomandibular joint arthroscope: development and preliminary clinical findings. AB - This study evaluated the development and preliminary results of a single-channel thin-fiber and Nd:YAG laser temporomandibular joint arthroscope. No articular damage from the arthroscopic procedure and laser lysis was observed in any of the joints when the arthrotomy was performed. All three disk perforations found by the arthroscope were confirmed at the time of the arthrotomy, and in these joints the tip of the arthroscope could be advanced into the inferior joint. The nine joints, where only arthroscopic laser lysis was performed, were followed up and the results were satisfactory. PMID- 9418143 TI - Interpositional bone grafting and Le Fort I osteotomy for reconstruction of the atrophic edentulous maxilla. A two-stage technique. AB - This study presents the results from ten consecutive patients who, because of insufficient bone volume for conventional implant placement in the maxilla, were treated with an interpositional bone graft and Le Fort I osteotomy. The endosteal implants were placed six months after the osteotomy. A total of 60 screw-shaped titanium implants (Branemark) were placed, of which three failed to integrate during the six-month healing period. No further implants were lost during the follow-up period, ranging from 15 to 39 months after placement of the implants. All patients received fixed bridges and all have continued to function efficiently. PMID- 9418144 TI - Bone grafting to the maxillary sinuses, nasal floor and anterior maxilla in the atrophic edentulous maxilla. A two-stage technique. AB - This study presents the results from 20 consecutive patients treated with an autogenous bone graft from the iliac crest. In ten patients the graft was placed in the maxillary sinuses and the floor of the nose (inlay group). Ten patients, in addition to the inlay graft, had a corticocancellous bone block secured with mini-screws to the anterior maxillary ridge (inlay/onlay group). Endosteal implants (Branemark) were placed six months after surgery. A total of 136 implants were placed, of which eight failed to integrate during the six-month healing period. A further 15 implants were lost during the follow-up period. For the inlay group the average follow-up period was 22 months and for the inlay/onlay group 19 months. Donor site morbidity was significantly less when iliac bone was harvested with a trephine (inlay group) than in patients treated with our routine procedure for bone harvesting (inlay/onlay group). Surgical technique, donor site morbidity, implant survival and patient acceptance are presented. PMID- 9418145 TI - Factors influencing survival rate in adenoid cystic carcinoma of the salivary glands. AB - Ninety-one cases of adenoid cystic carcinoma (ACC) of the salivary glands with more than ten years' follow up were studied to investigate factors influencing the survival rate of patients, which vary according to site, histological type, clinical stage and nature of therapy. The data were statistically analysed for survival curves. Log rank tests were employed to assess the statistical significance of various groups. As a result, it may be concluded that tumour site, clinical stage and histological type are the important factors influencing the prognosis. ACC of the palate and parotid, early clinical stage, glandular/tubular histological type, and tumour without nerve involvement had the best prognosis. ACC in the submandibular gland, maxillary antrum and tongue, advanced clinical stage (stage III and IV), solid histological type, and tumour with nerve involvement had a poor prognosis. PMID- 9418146 TI - "Chain-link" combined tissue transfer for the reconstruction of the mandible. AB - We describe a procedure for "chain-link" combined tissue transfer connecting the vascular pedicle of a deep circumflex iliac flap with that of a forearm flap after wide resection of the mandible. Combination of these flaps facilitated the reconstruction of the defect in both intra- and extraoral soft tissue and the mandibular bone. This method is useful when cervical recipient blood vessels are limited due to the wide resection of the primary tumor and radical neck dissection. PMID- 9418147 TI - CD44 variant 6 (CD44v6) expression as a progression marker in benign, premalignant and malignant oral epithelial tissues. AB - The change in the expression pattern of CD44 variant 6 (CD44v6) protein in benign, premalignant, and malignant (SCC) oral epithelial lesions was studied immunohistochemically and compared with the pattern in normal mucosa in order to examine whether this gene can serve as a progression marker in patients with SCC. The principal findings is that CD44v6 expression was clearly downregulated in most cases of severe premalignant lesions as well as in most of the SCCs. The staining pattern and intensity varied according to the degree of dysplasia and to the degree of differentiation of the SCCs. Premalignant severe epithelial dysplasia cases with early features of invasion, not yet developed into SCC, showed distinctly downregulated expression of CD44v6 protein whereas hyperplastic and benign epithelial lesions (papilloma) expressed positive staining patterns comparable to those of the normal counterparts. The authors conclude that alteration in CD44v6 may occur as an early event in primary oral SCC development, as well as in premalignant severe epithelial dysplasia. It can thus, be used as a molecular progression marker when screening for oral cancer. PMID- 9418148 TI - Regional metastasis in polymorphous low grade adenocarcinoma. Report of a case. AB - A case of cervical lymph node metastases of polymorphous low grade adenocarcinoma is reported, originating from a minor salivary gland in the soft palate in a 76 year-old man, 19 years after initial presentation. The clinical course, histopathology, and literature review on the metastatic aspect of the tumour are presented. This has emphasized the need for increased awareness of this recently described tumour and the need for lifelong follow up. PMID- 9418149 TI - Treatment of chronic suppurative osteomyelitis of the mandible. AB - Chronic suppurative osteomyelitis of the mandible is often considered difficult to treat and may lead to refractory osteomyelitis. Sixteen patients with chronic suppurative osteomyelitis of the mandible were treated with a relatively simple protocol, consisting of sequestrectomy or decortication and i.v. antimicrobial therapy for one week, followed by oral penicillin for three weeks. Only one case showed recurrence of symptoms, which was treated successfully in a second session. It is concluded that combined surgical and antimicrobial therapy should be sufficient to cure most cases of chronic suppurative osteomyelitis of the mandible. PMID- 9418150 TI - Ameloblastic fibrodentinoma. Report of two cases. AB - Two cases of ameloblastic fibrodentinoma localized in the posterior mandibular area are presented. The clinical and histological findings are discussed. PMID- 9418151 TI - Treatment of recurrent dislocation of the temporomandibular joint with type A botulinum toxin. AB - A case is reported of a 56-year-old woman who suffered from recurrent dislocations of the temporomandibular joint (TMJ) secondary to an exacerbated tetraspastic syndrome of multiple sclerosis. Following chemical denervation of the masseter and pterygoid muscles with injections of type A botulinum toxin, no further dislocations occurred for periods of up to four months. The treatment has been repeated five times. Some of the indications and possible adverse reactions to this therapy are discussed and comparisons made with other, conventional methods for managing recurrent dislocation of the TMJ. PMID- 9418152 TI - An experimental rabbit model for jaw-bone healing. AB - The purpose of this investigation was to study the structural and topographical bone anatomy of the right and left edentulous areas between the incisors and molars in the rabbit maxilla with regard to the symmetry of the bone, and to assess the degree of spontaneous healing of surgical defects. Anatomical and radiographic examinations together with analysis of serial histological ground sections in ten rabbits disclosed no statistically significant differences between the two sides regarding the different bone-tissue structures, i.e. they exhibit a sufficient degree of symmetry to serve as a useful bilateral test control model. Surgical defects were made on one side of the jaw (test side) in a group of eight rabbits. This resulted in an average loss of 17% of the total bone volume after a healing period of four weeks as compared to the untreated control side. It was concluded that surgically-created defects do not show completely spontaneous healing. From a histological section of the test side, it was possible to redraw the original bone contour by interpolation between unaffected areas of bone, coronal and apical to the defect. This means that the test side of this model can also serve as its own control with regard to the amount of regenerated tissue, given that there is unaffected bone, coronal and apical to the defect. PMID- 9418153 TI - Risk of HIV-1 transmission in heterosexual stable and random couples. The Italian Partner Study. PMID- 9418154 TI - Mother-to-child transmission of HIV-1: risk factors and prevention. PMID- 9418155 TI - Issues on antiretroviral post exposure combination prophylaxis. AB - Early antiretroviral chemoprophylaxis after HIV exposure can reduce the risk of transmission according to in vitro studies, animal experiments, and the results of a case control study on occupationally exposed health care workers. Additional consistent evidence comes from trials on treatment of acute infection and on prevention of vertical transmission. Combination post exposure prophylaxis (PEP) with available antiretroviral agents is recommended because of higher antiretroviral activity, and to overcome the increasing problem of resistant HIV strains. Recommendations include the addition of a second reverse transcriptase inhibitor antiretroviral agent to zidovudine for the majority of exposures and the addition of a protease inhibitor for, at least, highest risk exposures. Starting PEP as soon as possible is strongly recommended, within 1-4 hours from the exposure; the recommended duration is four-weeks. Contraindications are those specified in the package insert for each product. PEP guidelines refer to exposures occurring in health care settings. PEP can be considered in the case of rape or sexual exposure that occur in isolation in individuals committed to safe practices, as in the case of breakage of condoms in a discordant couple, or who intend to stop risk practices, as an intravenous drug user who comply to maintenance or detoxification program. PEP is not recommended in the case of injury with abandoned needles. Being the ideal PEP regimen not fully defined, there is the need to collect further information through the institution of National and International Combined PEP Registries. Because of its complexity PEP management needs high professional skills and service organisation. Moreover, the suggested guidelines can not be considered definite and post exposure policies and PEP protocols that reflect best updated practice should be ensured. PMID- 9418156 TI - The dynamics of human immunodeficiency virus type 1 transmission among injecting drug users. PMID- 9418157 TI - Heterogeneity in exposed uninfected individuals. AB - In spite of repeated exposures to HIV, some individuals remain seronegative and apparently uninfected. A variety of mechanisms potentially able to confer resistance to HIV infection, including cell-mediated and (unconventional) humoral immune responses, as well as mutations affecting receptors for virus entry have been considered and analysed. In this article, we want to discuss recent reports on specific immune responses and genetic factors potentially involved in mechanisms of protection, and to present some of our data relative to a cohort of people sexually exposed to HIV-1, but persistently seronegative. These EU (exposed uninfected) individuals can be distinguished from "normal" unexposed controls on the basis of significantly increased frequencies of a number of immunological parameters that might be considered "unconventional" correlates of HIV infection/protection. However, EU individuals are highly heterogeneous since the various unconventional immune responses considered can be present in all possible combinations. Aim of future research will be to ascertain the role of such immune responses in the maintenance of the protection state, or their secondary nature as signals of a particular kind of infection. PMID- 9418158 TI - Viral determinants in HIV-1 transmission. PMID- 9418159 TI - New mechanisms of viral persistence in primary human immunodeficiency virus (HIV) infection. AB - Viruses, including the Human Immunodeficiency Virus (HIV), have evolved multiple strategies to overcome host immune defenses, allowing them to persist in the host. Molecular and cellular approaches were simultaneously used to provide sensitive and unbiased delineation of the diversity and dynamics of the immune response, and to study the relative compartimentalization of HIV-specific CTL clones in patients undergoing primary HIV infection. This approach revealed that some HIV-specific CTL clones can be deleted in presence of high levels of antigen, a phenomenon analogous to high-dose tolerance or clonal exhaustion described in murine models of persistent viral infections. Also, HIV-specific CTL clones were found to accumulate preferentially in peripheral blood as compared to lymph nodes, even though the large majority of viral replication during primary HIV infection takes place within lymph nodes. These two mechanisms may decrease the effectiveness of the host cell-mediated immune responses, and favor the establishment of virus persistence during primary HIV infection. PMID- 9418160 TI - Increased plasma levels of the C-C chemokine RANTES in patients with primary HIV 1 infection. AB - To investigate the role played by chemokines in the natural history of human immunodeficiency virus (HIV) infection, we measured the plasma levels of RANTES. MIP-1 alpha and MIP-1 beta in a cohort of patients with primary HIV-1 infection (PHI) followed longitudinally. The cohort included 17 patients with well documented history of acute HIV syndrome within two months of the first observation. The mean plasma concentration of RANTES, but not that of MIP-1 alpha or MIP-1 beta, was significantly higher in patients with PHI (192.3 ng/ml) than in five HIV-seronegative controls (8.0 ng/ml) studied during the same time period. Treatment of blood with a cocktail of drugs preventing platelet activation, followed by high-speed centrifugation, reduced the levels of RANTES by approximately 2 logs both in patients and in controls, indicating that the bulk of RANTES was released by platelets, which are known to store this chemokine in their alpha-granules, in the immediate aftermath of blood drawing. No correlation was seen between the levels of RANTES and the number of HIV genome equivalents in plasma. These data suggest that large amounts of pre-formed RANTES are stored in platelets and, possibly, in other blood cells during the early phases of HIV infection. The possible role of this HIV-suppressive chemokine in the control of viral replication during PHI remains to be established. PMID- 9418161 TI - Soluble CD30, tumour necrosis factor (TNF)-alpha, and TNF receptors in primary HIV-1 infection: relationship with HIV-1, RNA, clinical outcome and early antiviral therapy. AB - The natural course of human immunodeficiency type 1 (HIV-1) infection varies considerably. The identification of laboratory disease markers has become critically important to patient management. This study, carried out on 37 patients with primary HIV-1 infection (PHI), shows that, along with plasma HIV-1 RNA and CD4+ T cell counts, evaluation of plasma levels of some immune activation markers (sCD30, TNF-alpha, and sTNFR-I) may help to identify patients at risk of a more rapid disease progression, suggesting that immune activation is among the factors who determine the rate of disease progression. Early combination antiviral therapy significantly decreased levels of virus load and of immune activation markers, suggesting that it may reduce the extent of immune activation through the suppression of HIV-1 replication. Among others, sCD30 could be a more sensitive marker of immune activation, and it might be also useful in the monitoring of the response to antiviral therapy. PMID- 9418162 TI - Cellular immunity. PMID- 9418163 TI - Primary HIV infection: clinical aspects and monitoring. PMID- 9418164 TI - Primary HIV infection: therapeutic strategies. PMID- 9418165 TI - Recent advances in antiretroviral therapy of HIV infection. PMID- 9418166 TI - Molecular monitoring of human immunodeficiency virus type 1 infection. AB - Over the past few years, considerable technical effort has been directed to developing molecular methods that would allow an effective approach to the diagnosis of human immunodeficiency virus type 1 (HIV-1) infection and its monitoring. Indeed, quantitative molecular techniques have opened the way for a new type of direct study of untreated and treated HIV-1 infected subjects. The understanding of the immunopathogenesis of HIV-1 infection has increased significantly with the introduction of advanced virological and molecular methods for accurate quantitative analysis of HIV-1 activity; powerful methodologies answer (directly and in real time) most questions generated by pathogenic research and by the novel anti-viral strategies introduced in clinical practice. The data from pilot diagnostic applications of quantitative techniques have clarified important features of the natural history of HIV-1 infection. Moreover, an increasing amount of data indicate the need for second-level laboratory facilities for the clinical management of infected patients; virological aspects and some genetic features of the hosts concerning HIV-1 co-receptors (all the co receptors so far identified are members of, or related to, the transmembrane, chemokine-receptor family) need to be elucidated for the complete diagnostic evaluation of HIV-1-infected subjects. PMID- 9418167 TI - HIV infection in macrophage: role of long-lived cells and related therapeutical strategies. AB - Therapeutical strategies aimed to the maximal inhibition (if not the eradication) of infection by human immunodeficiency virus should take into account the issue of the viral reservoir in the body. Recent data clearly show that latently infected lymphocytes represent a minimal part of the viral reservoir, while the majority of these cells are macrophages (variably differentiated) scattered in the tissues and lymph nodes. Immunologically-sequestred areas, such as the central nervous system, are particularly relevant in view of the different concentrations of antiviral drugs achieved in the organs. Thus, a careful analysis of the distribution of antiviral drugs, and the assessment of their activity in cells of macrophage lineage, represent key factors in the development of therapeutical strategies aimed to the "cure" of infectious patients. PMID- 9418168 TI - Experiences in immune reconstitution. The rationale for interleukin-2 administration to HIV-infected individuals. AB - Since the clinical earliest descriptions of patients with acquired immune deficiency syndrome (AIDS) it has been very clear that a profound state of immunologic dysfunction was the underlying cause of the emergence of life threatening opportunistic infections and tumors. In addition to the progressive loss of CD4 "helper" T lymphocytes, a profound defect in interleukin-2 (IL-2) production was recognized as a major pathogenic component of the new disease. For these reasons, attempts to administer IL-2 to individuals infected with the human immunodeficiency virus (HIV), the causative agent of AIDS, have been made since the mid eighties, however with little success. On the other hand, the propensity of HIV to replicate in activated lymphocytes and macrophages, under the influence of the cytokine network, has represented, and in part still does, a major hurdle for the rationale of administering IL-2 or other cytokines to HIV-infected individuals. Major steps forward towards an understanding of the role of multiple components of the immune system, coupled with a potentially successful protocol of IL-2 administration in vivo, resulting in the stable uprising of circulating CD4+ T cells, shed an optimistic light on the possibility to achieve a substantial immune reconstitution in HIV-infected individuals, thus preventing the onset of AIDS. PMID- 9418169 TI - An approach to HIV gene therapy by transduction of multifunctional retroviral vectors in primary human T lymphocytes. PMID- 9418170 TI - Vaccines against HIV. PMID- 9418171 TI - Self-talk in distressed youth: states-of-mind and content specificity. AB - Tested hypotheses derived from a States-of-Mind (SOM) model in a sample of 542 children ages 7 to 15. SOM ratios were computed using empirically determined items. Support was found for the SOM model. Negative self-talk (but not positive) was consistently related to increasing levels of affective distress. Thus, children comorbid for anxiety and depression demonstrated the highest levels of negative self-talk (and most dysfunctional SOM ratios). Support for content specificity (specific cognitive content relating uniquely to affective conditions) was mixed. However, specificity was increased when attention was paid to more rationally derived content. Age was not a predictor of positive or negative self-talk. Discussion considers the use of the SOM model in child psychopathology and psychotherapy research. PMID- 9418172 TI - The role of social anxiety in adolescent peer relations: differences among sociometric status groups and rejected subgroups. AB - Examined the relation between sociometric nominations and social anxiety in adolescence. Participants were 973 students (473 boys and 500 girls) in Grades 6, 7, 8, and 9. Students completed the Social Anxiety Scale for Adolescents and a sociometric nomination task that included the following behavioral descriptors: liked most, liked least, starts fights the most, best sense of humor, class leader, easiest to push around, and most cooperative. Sociometric nominations were used to classify students into standard sociometric status groups (i.e., popular, average, rejected, neglected, and controversial) as well as into rejected subgroups (aggressive rejected and submissive rejected). Results indicated that students classified as rejected and neglected reported more social anxiety than those classified as average, popular, or controversial. In addition, submissive rejected students reported significantly more social anxiety than did aggressive rejected or average students. Implications of these results for assessment and treatment of adolescents with peer problems are discussed. PMID- 9418173 TI - Diagnosing ADHD (predominantly inattentive and combined type subtypes): discriminant validity of the behavior assessment system for children and the achenbach parent and teacher rating scales. AB - Compared the effectiveness of discriminating attention deficit/hyperactivity disorder (ADHD) subtypes using the Parent Rating Scale (PRS) and Teacher Rating Scale (TRS) of the Behavior Assessment System for Children (BASC) and the Parent Report Form and Teacher Report Form (TRF) of the Achenbach Child Behavior Checklist (CBCL). To determine the extent to which these scales measured similar behaviors, Pearson Product-Moment Correlations were computed for the parent scales (PRS and CBCL) and for the teacher scales (TRS and TRF). Results indicated that correlations were significant for a number of scales. Discriminant analysis does not suggest a strong advantage of either measure in differentiating children with ADHD from those who do not meet criteria for ADHD, except for the BASC TRS which has better predictive ability for children who do not meet ADHD criteria. For subtypes of ADHD, and specifically the ADHD: Predominantly Inattentive subtype, however, results would favor the use of the BASC PRS and TRS. PMID- 9418174 TI - Fear of fat, disregulated-restrained eating, and body-esteem: prevalence and gender differences among eight- to ten-year-old children. AB - Examined whether: (a) societal directives to be thin are perceived among children, (b) discontent with body and attitudes and behaviors associated with eating disorders begin before adolescence, and (c) these differ by sex. These issues were assessed in 239 Grade 3 students. Scales of eating and weight attitudes and behaviors for this under-studied population were either created or modified from existing instruments. These 8- to 10-year-old children expressed weight, dieting, and physique concerns that reflect Western sociocultural values and preoccupation with body weight and dieting. Sex differences were examined and revealed several but not very reliable distinctions at this young age. These findings appear to be consistent with research on adolescents. The components that may lead to the development of an eating disorder or disregulated-restrained eating in a vulnerable adolescent may be both internalized and expressed at a very early age. PMID- 9418175 TI - Convergent and discriminant validity of measures of parenting efficacy and control. AB - Examined the convergent and discriminant validity of the Parent Attribution Test (PAT; Bugental, Blue, & Cruzcosa, 1989), the Parental Locus of Control Scale (PLOC; Campis, Lyman, & Prentice-Dunn, 1986), and the Parenting Sense of Competence-Efficacy Scale (PSOC-Efficacy; Johnston & Mash, 1989) in 3 samples of community mothers. In the 1st 2 samples, mothers also completed measures of negative affect and social desirability. In the 3rd sample, the PAT and PSOC Efficacy scales were administered with measures of adult attachment style and child behavior problems. There was weak support for the convergent validity of the measures. Moreover, the discriminant validity of the measures was not adequately demonstrated. Our results suggest that PLOC and PSOC-Efficacy scores may reflect distress and response style as well as beliefs about parenting. Scores on the PAT, although less influenced by response style and distress, appear to reflect a different dimension of efficacy than that assessed by other self-report measures. PMID- 9418176 TI - Effect of diagnosis on countertransferential responses to child psychotherapy patients. AB - Examined the impact of patient diagnosis on countertransferential responses to hypothetical latency-aged child psychotherapy patients meeting criteria for Dysthymia, Conduct, and Borderline Disorders. Three subtypes of countertransference (CT) were measured: positive (e.g., nurturant feelings), negative (e.g., boredom), and CT-related activity (e.g., tendency to refer patient to another therapist). All three subtypes were found to vary significantly as a function of diagnosis. Of the three diagnostic groups, Dysthymic patients elicited the highest degree of positive CT; Conduct Disorder patients, the highest degree of negative CT; and Borderline patients, the highest degree of CT-related activity. Neither patient and therapist gender nor therapist experience was significantly related to any of the CT scores; however, therapist psychological distress was found to be significantly associated with negative CT and CT-related activity for all three diagnoses. PMID- 9418178 TI - Global and domain-specific self-concepts of a matched sample of adolescent runaways and nonrunaways. AB - Assessed the self-concepts of 132 adolescents (66 adolescent runaways and 66 nonrunaways) with the Multidimensional Self Concept Scale (MSCS). Adolescent runaways were found to have significantly lower global self-concepts than the matched group of children who did not run away (hereafter nonrunaways). Runaways were also found to have lower domain-specific self-concepts than nonrunaways in 5 of the 6 important domains assessed by the MSCS (i.e., Social, Affect, Academic, Family, and Physical). Within-group analyses revealed that runaway adolescents had significantly lower Family self-concepts than other specific domains of self concept, which highlights the influence of families on children's psycho-social adjustment. As expected, nonrunaways did not differ across any of the domain specific self-concepts. PMID- 9418177 TI - A naturalistic study of psychotherapeutic methods and client in-therapy functioning in a child community setting. AB - Utilized a naturalistic methodology to examine treatment responses associated with major psychotherapeutic methods in 150 youth aged 11 to 17 years old in a community mental health center. Treatment methods were not experimentally controlled but were measured retrospectively by therapist report. Treatment response was assessed by a composite of 6 measures completed by clients, parents, and therapists. Pretest/posttest comparisons indicated improved functioning in the sample as a whole. Treatment response was not related to the proportion of therapy using individual, family, or group modalities. Therapy response was positively associated with extent of use of cognitive therapy. Social skills training, behavior therapy, and family systems therapy were associated with more positive treatment response in some subgroups of clients. The number of approaches used in an individual case (technical eclecticism) was positively related to client response. Treatment response was more consistently related to level of client and parent functioning in therapy than to treatment method. PMID- 9418179 TI - Empirical evaluation of DSM-IV generalized anxiety disorder criteria in children and adolescents. AB - Evaluated the Diagnostic and Statistical Manual of Mental Disorders (4th ed. [DSM IV], American Psychiatric Association, 1994) generalized anxiety disorder (GAD) criteria in children and adolescents. Clinic-referred children meeting criteria for DSM-IV GAD, those meeting criteria for another DSM-IV anxiety disorder, and normal children participated in a structured interview and completed self-report questionnaires. Groups were compared in terms of interview and self-report measures to examine convergent and discriminant validity. In addition, developmental differences, cross-informant symptom and syndrome agreement, and validity of parent and child report were determined. Finally, the symptoms comprising the GAD associated symptom criterion (Criterion C) were examined in terms of rate of endorsement and predictive power. Results showed that parameters of worry differentiated children with GAD from those with other anxiety disorders and controls. Developmental differences in the sample did not appear to necessitate a separate criteria set for the classification of generalized anxiety in children of this age. Symptoms from GAD Criterion C evidenced moderately high rates of endorsement and acceptable predictive power. Overall, the DSM-IV GAD criteria for children and adolescents are supported, but further evaluation is necessary before firm conclusions can be drawn. PMID- 9418180 TI - Marital adjustment, marital discord over childrearing, and child behavior problems: moderating effects of child age. AB - Examined whether marital discord over childrearing contributes to child behavior problems after taking into account general marital adjustment, and if child age moderates associations between child behavior problems and either general marital adjustment or marital discord over childrearing. Participants were 146 two-parent families seeking services for their child's (4 to 9 years of age) conduct problems. Data on marital functioning and child behavior problems were collected from both parents. Mothers' and fathers' reports of marital discord over childrearing related positively to child externalizing problems after accounting for general marital adjustment. Child age moderated associations between fathers' reports of general marital adjustment and both internalizing and externalizing child problems, with associations being stronger in families with younger children. The discussion highlights the role that developmental factors may play in understanding the link between marital and child behavior problems in clinic referred families. PMID- 9418181 TI - Assessment of peer neglect in the preschool years: a short-term longitudinal study. AB - Examined concurrent and longitudinal relations between different measures of peer neglect in the preschool years. Measures of social competence included peer sociometrics, teacher ratings, and behavioral observations of peer interactions. Participants were sixty 4- to 5-year-old Caucasian boys from low-income family backgrounds. Results indicated that the stability of indicators of peer neglect and social isolation depended on the measure employed. Correlations between different measures suggested heterogeneity in patterns of social adaptation among neglected or isolated preschool children. Results are discussed emphasizing the need to rely on multiple indicators for the assessment of preschool children's social competence. PMID- 9418182 TI - Strengthening child compliance through positive parenting practices: what works? AB - Examined the laboratory playroom behavior of 36 normal mother-child dyads during a competitive game in which the mothers gave periodic instructions to their children. The dyads were equally divided into a control group, and two experimental groups in which mothers were taught to use positive parenting in the form of mirroring or praise. The game activity was videotaped and observers coded rates of child compliance, maternal instructions, mirroring and praise, and the mothers' overall responsiveness to their children's full repertoire of behaviors. After the game, mothers and children were interviewed separately to assess their satisfaction with the game interaction. Results showed higher percentages of child compliance and higher ratings of dyadic satisfaction for the experimental groups. However, maternal responsiveness proved to be the only significant predictor of these dependent measures, suggesting that mirroring and praise are specific markers for a more complex parenting process. Implications of these results for parenting practices and parent training are discussed. PMID- 9418184 TI - Altered expression and function of P-glycoprotein (170 kDa), encoded by the MDR 1 gene, in T cell subsets from aging humans. AB - Aging is associated with progressive T cell-mediated immune deficiency, increased frequency of infections, and autoimmune phenomena. P-glycoprotein (P-gP), a 170 kDa glycoprotein, is a member of a superfamily of ATP-binding cassette transport proteins that has been shown to express on cells of the immune system and suggested to play a role in secretion of certain cytokines and cytotoxic molecules. Because aging is associated with altered secretion of cytokines, in this investigation we examined the expression and function of P-gP in CD4+ and CD8+ T cells and their "memory" and "naive" subpopulations in peripheral blood from healthy aging and young subjects. P-glycoprotein expression was analyzed at the protein levels by dual- or triple-color flow cytometric analysis, using monoclonal antibodies against P-gP (MRK16), and at the mRNA level by quantitative reverse-transcriptase polymerase chain reaction. The efflux function of P-gP was measured by intracellular accumulation of rhodamine-123 (Rh123; a substrate for P gP) in the presence or absence of cyclosporin A (which binds to P-gP and inhibits its efflux function). The data show increased expression of P-gP at both the protein and the mRNA levels in aging lymphocytes. Increased P-gP expression, at the protein level, was also observed in naive cell subpopulations from aging CD4+ and CD8+ T cell subsets compared to those from young controls. An increase in P gP function, as measured by the ability of T cell subsets to efflux Rh123, was observed in aging CD4+ and CD8+ T cell subsets and their naive and memory subpopulations. These data suggest that altered P-gP expression and function in aging may play a role in changes in immune response, including cytokine secretion, associated with human aging. PMID- 9418185 TI - Autoimmunity to polymorphonuclears: functional consequences of the binding of antibodies to membrane and cytoplasmic target antigens of polymorphonuclear leukocytes. AB - Antineutrophil autoantibodies reacting with cytoplasmic antigens are associated with various types of vasculitides, whereas antibodies reacting with neutrophil membrane antigens are mostly related to autoimmune neutropenias. The aim of this study was the investigation of the effect of monoclonal antibodies (MoAbs) reacting with surface and cytoplasmic antigens of polymorphonuclear leukocytes (PMN) known to be targets for autoantibodies in human diseases. Blood of healthy volunteers was tested for several phagocytic functions in the presence of MoAbs against surface (CD16, CD11b, CD18, NB1) and cytoplasmic (proteinase 3; PR3) molecules. Candidacidal activity was significantly inhibited in the presence of all MoAbs but isotypic control. Phagocytic activity was inhibited by anti-CD11b and/or anti-CD18 MoAbs. Zymosan-induced chemiluminescence was reduced by MoAbs anti-CD16, CD18, and NB1, enhanced by anti-PR3 MoAb, and less enhanced by anti CD11b. In conclusion, antimembrane antibodies diminished phagocytic functions at multiple steps; in contrast, anticytoplasmic MoAb promoted activation of oxidative burst in addition to impairment of microbicidal activity. This fact may be related to different pathogenic aspects of diseases associated with antimembrane and anticytoplasmic antibodies. PMID- 9418187 TI - Clinical trial of immunostimulation with a biological response modifier in unexplained recurrent spontaneous abortion patients. AB - We determined clinically whether a killed streptococcal preparation (KSP), a biological response modifier, is as effective as paternal lymphocyte immunotherapy for unexplained recurrent pregnancy abortion (RSA) therapy. The success rate of adverse pregnancy in the study group of 23 RSA cases, who were administered low doses of KSP before and during early pregnancy, was statistically compared with that in a control group of 205 women who received paternal lymphocyte immunotherapy. The association of natural killer (NK) cell activity in the peripheral blood with pregnancy outcome was also assessed. The success rate in the study group was 73.9% (17/23), compared to 75.1% (154/ 205) observed for the controls (not significant). Most of the successful cases exhibited low levels of NK cell activity in the peripheral blood. Immunotherapy with low doses of KSP is as effective as that with paternal lymphocytes, providing a simple and safe alternative therapy for unexplained RSA. Suppression of NK cell activity by some immunoregulatory mechanism was also found to have potential benefit in terms of a successful pregnancy outcome. PMID- 9418186 TI - Neutralization of transforming growth factor beta 1 augments hepatitis C virus specific cytotoxic T lymphocyte induction in vitro. AB - In hepatitis C virus (HCV) infection, TGF-beta 1 is upregulated in the liver and may be involved in the pathogenesis of chronic liver disease. TGF-beta 1 is also produced by activated T cells and acts as a potent immunosuppressor. The aim of this study was to investigate the roles of TGF-beta 1 in HCV-specific cytotoxic T lymphocyte (CTL) induction and enhance their killer activity by TGF-beta 1 modulation. We generated anti-HCV CTL from peripheral blood mononuclear cells from HLA-A2 patients under stimulation with the HCV-core peptide having the HLA A2.1 binding motif. The lytic activities of CTL or precursor frequency (CTLpf) generated with or without anti-TGF-beta antibody were compared. To optimize the IL-2 dose for CTL induction, low (50 U/ml) and high (500 U/ml) doses were tested and the lytic activities were compared. TGF-beta 1 amounts in the supernatants were assessed by enzyme-linked immunosorbent assay and by their growth inhibitory effect on mink lung epithelial cells. CTL activity was enhanced by anti-TGF-beta antibody in a dose-dependent manner but CTLpf did not significantly change. A high dose of IL-2 reduced the activity to 45% of that observed with a low dose, whereas TGF-beta 1 increased as the dose of IL-2 increased. Exogenous IL-10 reversed the inhibitory effect of a high dose of IL-2 on the killing activity by reducing TGF-beta 1 mRNA expression in T cells and its production. These results demonstrated that endogenous TGF-beta 1 is an autocrine suppressor in CTL induction in vitro. Therefore, the blockade of endogenous TGF-beta 1 could enhance the killing potential of anti-HCV CTL. PMID- 9418188 TI - Contribution of antibody to neutrophil-mediated killing of Enterococcus faecalis. AB - Late-onset septicemia due to Enterococcus faecalis is common among very low-birth weight neonates. These infants have low concentrations of placentally derived IgG and developmentally low levels of complement. The aim of the present study was to determine the contribution of antibody to in vitro neutrophil-mediated phagocytosis of E. faecalis. Antibody alone, as contained in an adult serum pool heated to inactivate complement, promoted only a modest reduction in the initial bacterial inoculum (50 +/- 12%) for 6 of 10 E. faecalis bacterial strains tested and allowed growth of the other four strains. In the presence of complement, NHS promoted > or = 90% reduction in the initial bacterial inoculum of two representative strains at serum concentrations as low as 0.5%. Hypogammaglobulinemic serum supported similar activity only at concentrations above 5%. Purification of IgG and IgM fractions from NHS revealed that IgM had the higher specific activity to promote phagocytic activity. Absorption to remove specific antibody significantly reduced bactericidal activity by normal human serum, complement-deficient sera, and hypogammaglobulinemic serum. Reconstitution of hypogammaglobulinemic serum with antibody as contained in 1% heated normal human serum or in immune globulin for intravenous use (1200 mg/dl) restored phagocytic activity. Thus, E. faecalis-specific antibody enhances PMN-mediated killing of this organism. Adjunctive therapy with intravenous immunoglobulin could augment the host response to enterococcal infections in infancy. PMID- 9418183 TI - Advances in the understanding of cytokine signal transduction: the role of Jaks and STATs in immunoregulation and the pathogenesis of immunodeficiency. AB - Cytokines are of great importance in the growth and differentiation of hematopoietic and other cells. Moreover, they are also crucial in immunoregulation and in host defense. Although our understanding of the molecular basis of cytokine action is far from complete, recent advances have substantially improved our knowledge of cytokine-dependent signal transduction. The delineation of the structure of cytokine receptors and the signaling pathways they utilize has provided clues as to how the strikingly specific effects of cytokines are achieved. Additionally, the basis of some of the pleiotropic and redundant effects of cytokines has also become clear. The discovery of the Janus family of protein tyrosine kinases (Jaks) and the STATs (signal transducers and activators of transcription) has also provided key insights into the mechanism by which intracellular signals are transduced. The following paradigm has emerged: cytokines induce dimerization of receptor subunits that are constitutively associated with Jaks. This activates the Jaks, which then phosphorylate the receptors. The phosphorylated receptors are bound by SH2-containing proteins, one class of which is the STATs. Activated STATs, then, translocate to the nucleus to effect gene transcription. Though the Jaks do not explain much in terms of specificity in signaling, the function of the STATs does. The discovery of patients with autosomal recessive severe combined immunodeficiency due to mutations of a particular Jak, Jak3, and the phenotype of knockout mice lacking Jak3 and various STATs demonstrate the specific and critical roles of these molecules in the development and function of the immune system. PMID- 9418189 TI - Epitope analysis of birch pollen allergen in Japanese subjects. AB - Birch pollen is a very common cause of nasal allergy (pollinosis) not only in Scandinavia, Europe, Canada, and the northern part of the United States but also in Hokkaido, Japan. We have previously reported a positive association between the HLA-DR9 phenotype and the development of birch pollen allergy in Japanese subjects. However, there is little information about T cell epitopes of birch pollen which are presented by HLA class II molecules other than HLA-DR9. Therefore, we analyzed the difference in T cell epitope usage in patients who had HLA-DR9 versus those who did not. Seven Japanese patients with birch pollinosis were studied. Some groups of peptides representing T cell epitopes (Betula verrucosa; Bet VI peptides, p7-33, p23-46, p138-160) appeared to be shared by the majority, while another peptide (Bet VI p72-95) was recognized predominantly by patients who expressed HLA-DR9 and/or HLA-DQ3 molecules. Moreover, seven T cell clones and eight T cell lines were generated from two patients who did not have HLA-DR9 or HLA-DQ3. Using some of these T cell clones/lines, we investigated the relationship between HLA class II molecules and antigenic peptides. One of these T cell clones recognized antigenic peptides in the context of the HLA-DQ1 molecule. To our knowledge, this is the first indication that the epitope on Bet VI can be presented by the HLA-DQ molecule. PMID- 9418190 TI - Regulation of ERK2 dephosphorylation in G1-stimulated rat T lymphoblasts. AB - Rat T lymphoblasts arrested in the G1 phase of the cell cycle by interleukin-2 (IL-2) deprivation can be forced to proceed to the S phase when they are stimulated with IL-2 or the phorbol ester phorbol 12,13-dibutyrate (PDBu). When PDBu is used as a stimulus, extracellular regulated kinase 2 (ERK2) is activated by threonine and tyrosine phosphorylation by the dual-specificity kinase MEK. Here we have studied the regulation of ERK2 dephosphorylation as a mechanism for inactivation of this kinase. In vivo inhibition of ERK2 dephosphorylation observed after preincubation with translation or transcription inhibitors (cycloheximide or actinomycin, respectively) indicates the involvement of at least one inducible phosphatase, the best candidate for which is the dual specificity phosphatase PAC-1. Other noninducible phosphatases must act as well, however, because sodium orthovanadate is a more effective dephosphorylation blocker than cycloheximide. In addition, the okadaic acid effect in ERK2 dephosphorylation indicates that Ser/Thr phosphatases are also involved, directly and/or indirectly. PMID- 9418193 TI - Histology and clinical significance of the carotid atherosclerotic plaque: implications for endovascular treatment. PMID- 9418191 TI - Detection of a soluble form of the leukocyte surface antigen CD48 in plasma and its elevation in patients with lymphoid leukemias and arthritis. AB - Proteins with glycosylphosphatidylinositol (GPI) anchors exhibit a range of activities and some of these proteins exist in both a membrane-associated and a soluble form. CD48 is a 47-kd GPI-linked glycoprotein which is expressed on T and B lymphocytes, monocytes, and many lymphoid malignancies. The biological function of CD48 is unknown. We describe the detection of a soluble form of CD48 in plasma and serum. Its level was quantified by an immunoenzymometric assay (IEMA) specific for soluble CD48. While soluble CD48 was detected in the plasma of healthy individuals (median = 29 ng/ml; range, 15-48 ng/ml), elevated levels were detected in some patients with lymphoproliferative disease (median = 41 ng/ml; range, 9-213 ng/ml, arthritis (median = 42 ng/ml; range, 13-67 ng/ml), and acute EBV infection (174 ng/ml). Soluble CD48 was also detectable in tissue culture supernatants from the Raji lymphoid cell line. The mechanism of CD48 release from cells is unclear. The finding of significant levels of soluble CD48 in plasma and the development of a sensitive IEMA for its measurement will facilitate further studies on its normal function and its role in disease. PMID- 9418194 TI - Endovascular stents for carotid artery occlusive disease. AB - PURPOSE: To study the feasibility and safety of endovascular stenting of cervical carotid artery stenosis. METHODS: Between April 1994 and May 1997, 108 consecutive patients (58 men; mean age 70.1 years) with > or = 70% carotid stenosis were treated with percutaneous stent implantation under a protocol that featured independent neurological review. Forty-four percent were asymptomatic. Over half the lesions (59%) were in the internal carotid artery; the mean stenosis was 86%. Palmaz stents were implanted without cerebral protection following preliminary balloon dilation; two Wallstents were used in long lesions. RESULTS: Carotid stents were successfully placed in 108 of 114 (95%) lesions. Of the 6 technical failures, 5 were access related and 1 was due to seizures during balloon dilation. Two major (1.8%) and 2 minor (1.8%) strokes occurred (3.7% stroke rate for 108 patients; 3.5% in 114 procedures), all in symptomatic patients, one of whom died. There were 5 (4.4%) transient ischemic attacks and 2 (1.8%) brief seizure episodes during dilation. One patient died of a cardiac event on day 20. The all stroke or death rate was 5.3% based on 114 arteries at risk (5.6% in 108 patients). In the mean 6-month follow-up (range 1 to 36) of 97 eligible patients, 3 (3.1%) died from unrelated causes. There was 1 restenosis (1.0%) from a stent compression, which was successfully redilated. There were no neurological sequelae, cranial palsies, or cases of stent or vessel thrombosis in follow-up. CONCLUSIONS: The use of stents in the treatment of cervical carotid occlusive disease appears feasible, effective in the short term, and without excessive risk of periprocedural stroke. PMID- 9418192 TI - A monoclonal antibody, DL10, which recognizes a sugar moiety of MHC class I antigens expressed on NK cells, NK+ T cells, and granulocytes in humans. AB - One mAb, DL10, was established from mice injected with dolphin lymphocytes. In addition to its reactivity against all dolphin lymphocytes, it reacted with some human leukocytes, including NK cells, NK+ T cells, and granulocytes. When its reactivity was examined in various animals, bovine, ovine, and equine leukocytes were DL10+. Murine, rat, and canine leukocytes were DL10-. Although the reactivity of DL10+ was similar to those of CD56 and CD57 antigens in humans, the actual molecules it recognized were different. Thus, all reactivity of DL10 disappeared after treatment of cells with glycopeptidase or after culture of cells with tunicamycin. Furthermore, the immunoprecipitation method revealed that DL10 indirectly recognized the heavy chain (45kD) of MHC class I antigen in humans and animals. Considering data from analysis of the N-terminal amino acid sequence of the DL10 molecule and the HLA typing of reactive cells, DL10 recognized a sugar moiety of some monomorphic MHC antigens and polymorphic MHC antigens such as HLA-B60 and -B61. If the donors are HLA-B60- and -B61 (> 80% in Japan and > 95% in the United States), DL10 would appear to be a very useful agent for the detection of pan-NK+ T cells. PMID- 9418195 TI - Traumatic carotid artery dissection and pseudoaneurysm treated with endovascular coils and stent. AB - PURPOSE: To report a case of post-traumatic internal carotid artery dissection and pseudoaneurysm formation at the C-1 level successfully treated by a percutaneous endovascular technique. METHODS AND RESULTS: A 20-year-old female presented 72 hours after a motor vehicle accident with incomplete occulosympathetic paresis (Horner's syndrome), carotidynia, and leftsided 1.5-cm x 2.5-cm pseudoaneurysm at the C-1 level. Neuroradiologists embolized the pseudoaneurysm with Guglielmi detachable coils and controlled the dissection with placement of a Wallstent. CONCLUSIONS: This report illustrates successful percutaneous endovascular treatment of a carotid dissection and pseudoaneurysm near the base of the skull. PMID- 9418196 TI - Endograft repair of an aortic pseudoaneurysm following gunshot wound injury: impact of imaging on diagnosis and planning of intervention. AB - PURPOSE: To describe the endovascular treatment of a gunshot injury to the visceral aorta and the role of various imaging modalities in the staging and planning of the endograft procedure. METHODS AND RESULTS: The bullet entered the aorta posteriorly beneath the origin of the superior mesenteric artery and traversed the wall in a tangential manner entering the lumen proximal to the renal arteries. Intravascular ultrasound (IVUS) imaging and spiral computed tomography (CT) identified the injury that the initial angiograms failed to demonstrate. Combined use of IVUS and CT imaging enabled observation of the evolution of a pseudoaneurysm until an interval when endograft exclusion was possible. A stent-graft was customized based on precise IVUS and CT dimensional data and implanted successfully through an arteriotomy in the common femoral artery 3 weeks after the initial injury. Three-month follow-up imaging demonstrated continued exclusion of the pseudoaneurysm, and the patient remains well at 16 months. CONCLUSIONS: IVUS and spiral CT scans were instrumental in identifying an arteriographically undetected aortic injury. The combined imaging modalities also helped determine the timing for the endovascular procedure and provided the precise measurements for device fabrication and deployment. PMID- 9418197 TI - Endografting in aortic trauma: let's keep it in perspective. PMID- 9418199 TI - Inflammatory reactions to endografts: The causes remain obscure. PMID- 9418198 TI - Acute inflammatory reaction associated with endoluminal bypass grafts. AB - PURPOSE: Nonspecific inflammatory reactions characterized by local tenderness, fever, and flu-like discomfort have been seen in patients undergoing endoluminal graft placement in the abdominal aorta or the femoral arteries. We undertook a study to assess the clinical and laboratory parameters of this inflammation. METHODS: Ten patients with femoropopliteal artery (n = 9) or aortic (n = 1) lesions were treated with EndoPro System 1 stent-grafts made of nitinol alloy and covered with a polyester (Dacron) fabric. Eleven patients implanted with a bare nitinol stent served as the control group. RESULTS: In the stent-graft group, four patients showed clinical signs of acute inflammation manifested by fever and local tenderness. Three of these patients suffered thrombosis of the stent-grafts during the first month of follow-up. Plasma levels of interleukin-1 beta and interleukin-6 in all stent-graft patients were markedly increased 1 day after intervention (7.3 +/- 2.8 versus 90.2 +/- 34.1 pg/mL and 15.6 +/- 5.8 versus 175.5 +/- 66.3 pg/mL, respectively; p < 0.01). This was followed by an increase in fibrinogen (3.0 +/- 0.2 versus 5.0 +/- 0.2 g/L; p < 0.05) and C-reactive protein (14.6 +/- 3.3 versus 77.5 +/- 15.0 mg/L; p < 0.01) at 1 week. No direct correlation between the inflammatory markers and symptoms could be found. In vitro analysis showed that individual components of the stent-graft did not activate human neutrophils, whereas the intact stent-graft itself induced a marked neutrophil activation. CONCLUSIONS: The component of the self-expanding stent-graft responsible for the nonspecific inflammatory reaction was not identified in this study. It is likely that the stent-graft itself or some as yet unrecognized element of the device other than the Dacron fabric or metal alloy may be a potent in vivo inducer of cytokine reaction by neutrophils. PMID- 9418200 TI - Three-year experience with modular stent-graft devices for endovascular AAA treatment. AB - PURPOSE: To evaluate feasibility and present early results of endovascular abdominal aortic aneurysm (AAA) exclusion using modular stent-grafts. METHODS: In a 3-year period ending July 1997, 201 patients were treated with self-expanding stent-grafts for AAAs with infrarenal necks > or = 10 to 15 mm long and < or = 32 mm wide; subtotal mural thrombus, calcification, and even angulation to some extent were acceptable, as were iliac arteries up to 18 mm wide. The patients were treated with either the Stentor/Vanguard device (178 cases) or the Talent endograft (23 cases). Follow-up on all patients was conducted at 3, 6, 12, 18, and 24 months. RESULTS: The technical aneurysm exclusion rate was 89% (178/201). There were 18 primary endoleaks (9.0%; 2 proximal, 16 distal), 4 (2.0%) conversions to open surgery, and 1 (0.5%) failure to deploy the graft. Seven (3.5%) patients died in the perioperative period, 5 due to multiorgan failure early in the series and two of hemorrhagic complications. Five (2.5%) renal artery occlusions were encountered; in one case, the graft was removed after 3 weeks. Nineteen late endoleaks were found in follow-up, related primarily to the iliac limb graft extensions of the Stentor device, graft material problems, or unknown causes. To date, 10 primary and 13 secondary endoleaks have been treated endovascularly. Twenty (10.0%) graft-limb thromboses were treated either by thrombolysis, thrombectomy, or a femorofemoral bypass. CONCLUSIONS: Endovascular grafting is technically feasible and becomes easier with improvements of the introducer systems and the grafts. The seemingly high complication rate in this series is due to the liberal patient selection criteria. PMID- 9418201 TI - Percutaneous endovascular stent-graft repair of iliac artery aneurysms. AB - PURPOSE: To report a percutaneous technique for endovascular repair of iliac artery aneurysms using commercially available materials. METHODS: Ten patients (9 males; mean age 65 +/- 11 years) presented with 11 isolated iliac artery aneurysms; 3 patients were asymptomatic. Stent-grafts were customized for each patient from polytetrafluoroethylene grafts with Palmaz stents sutured at either end of the tube. The devices were delivered percutaneously through standard 14F sheaths and deployed by balloon dilation. RESULTS: All iliac aneurysms were excluded without procedural incident. One patient with chronic renal insufficiency (baseline serum creatinine 1.9 mg/dL) experienced transient contrast-induced renal failure inhospital. The average hospital stay was 2.5 days (range 1 to 7). One vessel thrombosed 2 weeks following the procedure; the culprit stenosis at the site of arterial cannulation was dilated. One patient died of myocardial infarction at 6 weeks. The remaining eight grafts are patent and free of endoleak at a mean 14-month follow-up. CONCLUSIONS: This percutaneous technique appears to be an acceptable alternative to open surgical repair. PMID- 9418202 TI - Transcatheter embolization of a ruptured superior gluteal artery aneurysm: case report and review of the literature. AB - PURPOSE: To report the successful coil embolization of a true gluteal artery aneurysm and review therapeutic options for this rare condition. METHODS AND RESULTS: A ruptured superior gluteal artery aneurysm in a symptomatic 80-year-old man was successfully thrombosed by embolization using a combination of Gianturco coils and helical platinum microcoils. Six-month computed tomography demonstrated persistent thrombosis of the aneurysm and resolution of the perivascular blood. CONCLUSIONS: This report offers support for the use of catheter-based techniques as an alternative to standard surgical repair of gluteal artery aneurysms. PMID- 9418203 TI - Intravascular stenting in the superior mesenteric artery for chronic abdominal angina. AB - PURPOSE: Abdominal angina is an early clinical expression of occlusive mesenteric arterial insufficiency, a condition that requires aggressive treatment to prevent intestinal infarction. We report a case of chronic mesenteric ischemia in a young polyvascular man who had symptoms of abdominal angina. METHODS AND RESULTS: An aortic angiogram revealed a significant ostial stenosis of the superior mesenteric artery (SMA) associated with an occlusion of the inferior mesenteric artery. After predilation of the ostial portion of the SMA, significant residual stenosis remained. A balloon-expandable Palmaz P154 stent was deployed, restoring adequate luminal dimensions and blood flow. The patient was discharged after 2 days and remains asymptomatic at 5 months. CONCLUSION: Intraluminal stenting for treatment of mesenteric ischemia represents a viable alternative to surgical revascularization in selected cases. PMID- 9418204 TI - Recurrence of pseudoaneurysm after successful embolization. AB - PURPOSE: To report the initial successful treatment of a hepatic pseudoaneurysm by a partially formed coil and subsequent recurrence secondary to coil migration and configuration change. METHODS AND RESULTS: A 22-year-old man suffered a gunshot wound in the abdomen; a grade 4 liver laceration was identified and repaired. Eight days later, abdominal pain developed, and pseudoaneurysms were noted off both the superior and inferior branches of the right hepatic artery. Coil embolization was successful in occluding both defects; however, the inferior branch coil was incompletely formed. Twenty days later, symptom recurrence prompted angiography. The inferior branch coil had changed position and configuration, resulting in a larger pseudoaneurysm. Repeat embolotherapy was successful. CONCLUSIONS: The possible sequelae to maldeployed occluding coils must be considered even if the procedure appears successful. It may be advisable to place additional coils to more confidently occlude the pseudoaneurysm. PMID- 9418205 TI - Aortomonoiliac endografting: it doesn't have to be that difficult... PMID- 9418206 TI - Transmission electron microscopy and scanning force microscopy of poly r(A-U) and poly r(A-U)-ethidium bromide. AB - Transmission electron microscopy and scanning force microscopy of negative stained, carbon-coated replica and mica-adsorbed preparations of 200 microM poly r(A-U) and 50 microM ethidium bromide/200 microM poly r(A-U) have been employed to evaluate ethidium-induced changes in poly r(A-U) topology. Poly r(A-U) alone exhibits elongated conformations 85-115 nm in length that possess a number of hairpin loops as well as single-stranded domains. While the double-stranded domains are found predominately at the base of the hairpin loops (diameter = 5-30 nm), other rod-like (presumably double-stranded) regions ranging from 25-80 nm in length are present in other portions of the poly r(A-U). In contrast with the poly r(A-U) alone, the EB/poly r(A-U) combination appears as a heterogeneous population of condensed structures whose lengths and widths vary from 12-88 nm and 15-45 nm, respectively. These conformational changes are due to a number of factors, including the displacement of ordered water surrounding the poly r(A-U) and charge shielding of the phosphate groups of the poly r(A-U) upon the binding of the ethidium. PMID- 9418207 TI - Theoretical explanation of the relationship between backscattered electron and x ray linear attenuation coefficients in calcified tissues. AB - X-ray absorption and backscattered electron (BSE) microscopies are two commonly used techniques for estimating mineral contents in calcified tissues. The resolution in BSE images is usually higher than in x-ray images, but due to the previous lack of good standards to quantify the grey levels in BSE images of bones and teeth, x-ray microtomography (XMT) images of the same specimens have been used for calibration. However, the physics of these two techniques is different: for a specimen with a given composition, the x-ray linear attenuation coefficient is proportional to density, but there is no such relation with the BSE coefficient. To understand the reason that this calibration appears to be valid, the behaviour of simulated bone samples was investigated. In this, the bone samples were modelled as having three phases: hydroxyapatite (Ca10(PO4)6(OH)2), protein, and void (either empty or completely filled with polymethylmethacrylate (PMMA), a resin which is usually used for embedding bones and teeth in microscopic studies). The x-ray linear attenuation coefficients (calculated using published data) and the BSE coefficients (calculated using Monte Carlo simulation) were compared for samples of various phase proportions. It was found that the BSE coefficient correlated only with the x-ray attenuation coefficient for samples with PMMA infiltration. This was attributed to the properties of PMMA (density and mean atomic number) being very similar to those of the protein; therefore, the sample behaves like a two-phase system which allows the establishment of a monotonic relation between density and BSE coefficient. With the newly developed standards (brominated and iodinated dimethacrylate esters) for BSE microscopy of bone, grey levels can be converted to absolute BSE coefficients by linear interpolation, from which equivalent densities can be determined. PMID- 9418208 TI - Synthesis and in vivo evaluation of 7-chloro-5-[123I]iodo-4-oxo-1,4 dihydroquinoline-2-carboxylic acid. AB - As a possible radioligand for SPECT visualization of the NMDA receptors in the central nervous system, 7-chloro-5-[123I]iodokynurenic acid was prepared. This paper presents the synthesis of both the radioactive and the non-radioactive product, starting from 5-bromo-7-chlorokynurenic acid and using a non-isotopic nucleophilic halogen exchange reaction in the presence of iodide (Na123I or KI). Under the best labelling conditions, the radiochemical yield was 85%. The specific activity based on UV detection was found to be higher than 1 Ci/mumol (= 37 GBq/mumol) and the chemical and radiochemical (> 95%) purity of the tracer was checked by RP-HPLC. PMID- 9418209 TI - Single-step synthesis of [18F]haloperidol from the chloro-precursor and its applications in PET imaging of a cat's brain. AB - We have established a convenient synthesis process for the synthesis of[18F]haloperidol using a single-step 18F-for-Cl exchange reaction and a new elution system for the preparative high performance liquid chromatography (HPLC) using C18 bonded vinylalcohol copolymer gel (ODP) and a basic eluent. We successfully applied the product to cat-PET study and got clear images of the striatum, showing the usefulness of this synthesis. PMID- 9418210 TI - Synthesis of tritium-labelled N tau-methylhistamine for the improvement of extraction efficiency of N tau-methylhistamine from biological fluids. AB - In order to trace the loss of N tau-methylhistamine, a principal metabolite of histamine, during extraction and purification from human plasma and urine samples, N tau-[3H]methylhistamine was prepared in two steps from N alpha t butoxycarbonylhistamine (II). In the first step, compound II was deprotonated with NaH in an aprotic solvent and treated with [3H]methyl iodide. The products, N alpha t-butoxycarbonyl-N tau-[3H]methylhistamine (III) and N alpha t butoxycarbonyl-N pi-[3H]methylhistamine (IV), were then hydrolysed with iodotrimethylsilane under mild and short reaction conditions. Facile purification with Sep-Pak silica cartridges gave the combined two isomers of N tau [3H]methylhistamine and N pi-[3H]methylhistamine in 10.7% radiochemical yield with a radiochemical purity of > 94% and a ratio of approximately 2:1. Improvements in the extraction of methylhistamine using chromatography on Sep-Pak silica cartridges led to an overall recovery of 82.5 +/- 0.3% (n = 3) based upon total [3H]methylhistamine from normal human plasma. PMID- 9418211 TI - Evaluation of external dose equivalent with thermoluminescent dosimeters from residents living in radiation-contaminated buildings. AB - As of October 1996 there are more than 90 radiation-contaminated steel supported rebar buildings (containing more than 1000 apartments) dispersed in the northern part of Taiwan. These apartments were contaminated with cobalt-60 at a total activity ranging from 1-140 microSv/yr. In this paper, a method is developed for evaluating external dose equivalent and dose equivalent rates encountered by the residents wearing specially designed thermoluminescent dosimeter (TLD)-embedded chains, belts and badges. Comparisons are also made between the TLD readings and the exposure readings from indoor layout personal dosimetry surveys and room occupancy adjustments to the buildings. The accuracy and sensitivity of the TLDs compared with the ionization chamber readings are judged to be considerable improvements over those of previous studies. From the present study, it is concluded that the reliability of the daily activity records provided by the residents during the entire TLD-wearing period is the most critical but challenging feature of the external dose equivalent measurement. PMID- 9418212 TI - Activity concentrations and population dose from radium-226 in food and drinking water in Taiwan. AB - The purpose of this study was to determine the radioactivity of 226Ra in environmental samples in Taiwan. Fish, pork, rice, flour, chicken, vegetable, milk, fruit, egg and water samples were collected and pretreated by radiochemical procedure to extract the 226Ra, and the activity concentrations of 226Ra were determined using a liquid scintillation counter. The 226Ra content of groundwater was 12.0 mBq l-1. The 226Ra contents of the food ranged from 0.02 Bq kg-1 fresh to 0.17 Bq kg-1 fresh. The annual internal dose from ingestion of 226Ra from food and drinking water per caput was evaluated to be 7.5 microSv. PMID- 9418213 TI - Collimated detector technique for measuring a 137Cs deposit in soil under a clean protected layer. AB - A method to determine the 137Cs soil deposit in situ where the contamination is covered by a clean soil layer is described. The method is based on measurements of the count rates in three energy ranges by a collimated spectrum sensitive detector and their processing. The deposit determination accuracy is 20%. The measurement results are presented and compared with data from soil sampling. The 137Cs contamination maps of areas in Belarus and the Chelyabinsk region of Russia are presented. Measurements of the river Techa bank contamination detected the presence of secondary contamination. This method permits the change of dose rate after land decontamination and the work needed for decontamination and its efficiency to be predicted. PMID- 9418214 TI - Visual space distortion. AB - We are surrounded by surfaces that we perceive by visual means. Understanding the basic principles behind this perceptual process is a central theme in visual psychology, psychophysics, and computational vision. In many of the computational models employed in the past, it has been assumed that a metric representation of physical space can be derived by visual means. Psychophysical experiments, as well as computational considerations, can convince us that the perception of space and shape has a much more complicated nature, and that only a distored version of actual, physical space can be computed. This paper develops a computational geometric model that explains why such distortion might take place. The basic idea is that, both in stereo and motion, we perceive the world from multiple views. Given the rigid transformation between the views and the properties of the image correspondence, the depth of the scene can be obtained. Even a slight error in the rigid transformation parameters causes distortion of the computed depth of the scene. The unified framework introduced here describes this distortion in computational terms. We characterize the space of distortions by its level sets, that is, we characterize the systematic distortion via a family of iso-distortion surfaces which describes the locus over which depths are distorted by some multiplicative factor. Given that humans' estimation of egomotion or estimation of the extrinsic parameters of the stereo apparatus is likely to be imprecise, the framework is used to explain a number of psychophysical experiments on the perception of depth from motion or stereo. PMID- 9418215 TI - Use of the fractal dimension for the analysis of electroencephalographic time series. AB - Electroencephalogram (EEG) traces corresponding to different physiopathological conditions can be characterized by their fractal dimension, which is a measure of the signal complexity. Generally this dimension is evaluated in the phase space by means of the attractor dimension or other correlated parameters. Nevertheless, to obtain reliable values, long duration intervals are needed and consequently only long-term events can be analysed; also much calculation time is required. To analyse events of brief duration in real-time mode and to apply the results obtained directly in the time domain, thus providing an easier interpretation of fractal dimension behaviour, in this work we optimize and propose a new method for evaluating the fractal dimension. Moreover, we study the robustness of this evaluation in the presence of white or line noises and compare the results with those obtained with conventional spectral methods. The non-linear analysis carried out allows us to investigate relevant EEG events shorter than those detectable by means of other linear and non-linear techniques, thus achieving a better temporal resolution. An interesting link between the spectral distribution and the fractal dimension value is also pointed out. PMID- 9418216 TI - Timing of neural commands: a model study with neuronal networks. AB - Networks constructed of biologically realistic model neurons (neuroids) were used to study how in a neural assembly using pulse (interval)-coded information slow rhythmical oscillations with possible mode transitions might occur and how the efferent commands might be structured and their phase-shifts created. The simulations show that slow oscillations (in the hertz range) can be derived from reverberatory spiking in relatively short closed loops (fewer than ten neuroids) with the inputs protected against disturbing afferent signals and the outputs coupled by convergence on a common neuroid. Slow oscillations can be modified by a tonic activity entering the network; this activity changes the transmission time in the coupled loops involved. The structuring of the regulatory commands (in the millisecond range) was achieved by simulation of sequential activity propagation in a non-ring neuronal assembly supervised by a tonic activity in a set of inputs. The tonic activity acted as an instructive signal influencing the pattern of the functional connectivity in such a way that a particular efferent command was generated by the instructed network. PMID- 9418217 TI - Thinking about fantasy: are children fundamentally different thinkers and believers from adults? AB - Young children are often viewed as being unable to differentiate fantasy from reality. This article reviews research on both children's and adults' beliefs about fantasy as well as their tendency to engage in what is thought of as "magical thinking." It is suggested that children are not fundamentally different from adults in their ability to distinguish fantasy from reality: Both children and adults entertain fantastical beliefs and also engage in magical thinking. Suggestions are offered as to how children and adults may differ in this domain, and an agenda for future research is offered. PMID- 9418218 TI - Further distinctions between magic, reality, religion, and fiction. AB - Children's representations of counterintuitive phenomena can be better understood if we take into account the following: (1) Children may develop a conceptual slot for "counterintuitive + real" phenomena. (2) Notions of "reality" in early childhood are linked to experience rather than ontological status. (3) We have no good description of children's handling of fiction. (4) Cultural systems of religious representations make particular demands on developmental processes. PMID- 9418219 TI - The role of creative control and culture in children's fantasy/reality judgments. AB - Young children's ability to differentiate fantasy from reality has been seriously underestimated because of methodological problems and overgeneralization from children's performance in situations in which they had no control over the content of the fantasy and/or were presented with misleading information. It is important to keep in mind that there are many types of fantasy-reality distinctions, and that cultural context plays an important role in the interpretation given to children's activities. PMID- 9418220 TI - The last of the magicians? Children, scientists, and the invocation of hidden causal powers. AB - It is often assumed that scientific thinking displaces magical thinking, both historically and ontogenetically, but the tendency to invoke hidden causal powers straddles both modes of thought. The strength of this disposition among children and adults alike stands in need of explanation. PMID- 9418221 TI - Rescuing magical thinking from the jaws of social determinism. AB - Although there is otherwise much to recommend it, by riveting attention too narrowly on the contents of magical thought, and by recasting what is left of process in exclusively substantiative terms, this target article works to create the unwarranted impression that the magical thoughts of children and adults are all of a common piece. This commentary reads these oversights and omissions as symptoms of an unspoken new-situationalism working behind the back of Woolley's review. PMID- 9418222 TI - Crazy children, fantastical theories, and the many uses of metaphysics. AB - Woolley rightly challenges the incredible idea, held by some adults, that it is children who are peculiarly "fantastical" in their thinking. However, Woolley expresses little appreciation for "fantastical thinking" as it underlies the capacity for both grand delusions and amazing insights. In reducing "fantastical thinking" to conceptual error, she overlooks the mythical underpinnings of her own theorizing and neglects the many constructive roles of "fantastical thinking" in development. PMID- 9418223 TI - Human fetal heart rate dishabituation between thirty and thirty-two weeks gestation. AB - Few studies of human fetal habituation have included dishabituation procedures (i.e., assessment of the reemergence of a habituated response) to determine if response decrements are the result of reevaluation of information (a brain process) or fatigue of peripheral receptors. The purpose of this study is to describe the ability of the human fetus to learn and recall information with procedures to assess the central nervous system. Fetal heart rate (FHR) of 84 fetuses between 30 and 32 weeks gestational age was examined in response to 3 series of vibroacoustic (VA) stimuli presented at pseudorandom intervals of 25-45 s over the head of the fetus. Responses to the first series of 15 stimuli (S1) were compared with an identical second series of 15 stimuli (S1) presented over the head of the fetus. Between the 2 series, a novel (dishabituating) VA stimulus (S2) was presented, differing from S1 in intensity and frequency. The third series of S1s was applied to the mother's thigh as a control for possible maternal responses to the stimulus. Prestimulus FHR was computed during a 5 s interval before each stimulus, and mean FHR was computed during the intertrial interval (average FHR). The response to S1 during the first series of trials (1 15) produced a sustained rise in both prestimulus and average FHR, r(83) = .90, p < .001. After the novel S2 (trial 16) the rate of change was attenuated for average FHR, r(83) = .12, ns, to S1 for trials 17-31 but not prestimulus FHR, r(83) = .50, p < .001. The decrease in FHR response was reestablished when stimulation was applied to mother's thigh, trials 32-41, r(83) = .92, p < .001. A significant habituation pattern across trials was observed for the first series of S1s when prestimulus HR was subtracted from each preceding average FHR value (delta FHR). After the single novel stimulus (S2), the FHR response to S1 reemerged. All combinations of beginning and ending series slopes were compared, and only the rate of change during the last 4 trials of the initial presentation of S1 and the first 4 trials after the novel stimulus was significant, F(1, 82) = 9.21, p < .003. Uterine contractions collected from the continuous record were not related to the presentation of the novel stimulus, chi 2(1, N = 84) = 0.59, p < .50, ns, or delta FHR slope after the novel stimulus, chi 2(9, N = 84) = 10.52, p < .50, ns. These results established that the 32 week human fetus is capable of detecting, habituating, and dishabituating to an external stimulus and support the premise that areas of the human fetal central nervous system critical for detecting and discriminating information and for learning and memory have developed by the early third trimester. PMID- 9418224 TI - Extended visual fixation in young infants: look distributions, heart rate changes, and attention. AB - Visual fixation in infants from 3 to 6 months of age was examined for its fit to the theory of "attentional inertia." This theory posits that during the progression of a look there is increasing attention toward the stimulus and an "inertia" to continue looking. An extended audiovisual stimulus was presented for 20 min to infants while fixation was videotaped and heart rate (HR) was recorded. Consistent with the attentional inertia theory, look duration toward the stimulus had a lognormal distribution. Hazard functions describing these distributions showed a decreasing conditional probability of looking away with increases in look duration. Look onset and stimulus changes that occurred within a look were accompanied by HR deceleration. The average HR level continued to decrease over the duration of a look and returned to prestimulus level immediately prior to the fixation offset. Infant fixation has characteristics similar to fixation in children and adults, and attention appears to increase over the course of a look in young infants. PMID- 9418225 TI - Auditory context and memory retrieval in young infants. AB - Three-month-old infants were trained to move an overhead crib mobile while 1 of 2 musical selections was played. Retention was assessed 1 or 7 days later in the presence of either the same music or a different musical selection. In Experiment 1, the musical selections were very different (classical versus jazz); in Experiment 2, they were much more similar (two classical pieces). Infants in both experiments displayed 1 day retention regardless of which music was played during the retention test. At 7 days, retention was seen only when the music played during the retention test matched the training music. These data are consistent with similar findings showing that 3-month-old infants' memory is disrupted at long retention intervals when the context present during retention testing does not match the learning context. As the infant's memory wanes, context appears to function as a necessary cue for the retrieval of acquired expectancies. PMID- 9418226 TI - Comprehension of novel communicative signs by apes and human children. AB - Forty-eight young children (2.5 and 3.0 years old) and 9 great apes (6 chimpanzees and 3 orangutans) participated in a hiding-finding game. An adult human experimenter (the Hider) hid a reward in 1 of 3 opaque containers aligned on a wooden plank. Another adult experimenter (the Communicator) attempted to help the subject find the reward by giving 1 of 3 types of communicative sign: (1) Pointing, for which she placed her hand directly above the correct container with index finger oriented down; (2) Marker, for which she placed a small wooden block on top of the correct container; and (3) Replica, for which she held up a perceptually identical duplicate of the correct container. At both ages, children were above chance in this finding game with all 3 types of communicative sign, with Pointing being easiest (because they knew it prior to the experiment), Marker being next easiest, and Replica being most difficult. In contrast, no ape was above chance for any of the communicative signs that it did not know before the experiment (some had been trained in the use of the marker previously, and one knew pointing), nor was group performance above chance for any of the signs, despite the fact that apes experienced three times as many trials as children on each sign. Our explanation of these results is that young children understand the communicative intentions of other persons--although they may have more difficulty comprehending the exact nature of those intentions in some cases--whereas apes treat the behavioral signs of others as predictive cues only (signals). This may be because apes do not perceive and understand the communicative intentions of others, at least not in a human-like way. PMID- 9418227 TI - Preschoolers' understanding of the link between thinking and feeling: cognitive cuing and emotional change. AB - In 3 studies we investigated 3- through 6-year-olds' knowledge of thinking and feeling by examining their understanding of how emotions can change when memories of past sad events are cued by objects in the current environment. In Study 1, 48 4-, 5-, and 6-year-olds were presented with 4 illustrated stories in which focal characters experience minor sad events. Later, each story character encounters a visual cue that is related to one of his or her previous sad experiences. Children were told that the character felt sad, and they were asked to explain why. Study 1 suggested considerable competence as well as substantial development in the years between 4 and 6 in the understanding of the influence of mental activity on emotions. Studies 2 and 3 more systematically explored preschoolers' understanding of cognitive cuing and emotional change with different types of situations and cues. Across these 2 studies, 108 3-, 4-, and 5-year-olds listened to illustrated stories that featured story characters who each experienced a sad event and who were later exposed to a related cue. Children were not only asked to explain why the characters suddenly felt sad, but in some stories, they were also asked to predict and explain how another character, who was never at the past sad event, would feel. Results of Studies 2 and 3 showed an initial understanding of cognitive cuing and emotion in some children as young as 3, replicated and extended the evidence for significant developmental changes in that understanding during the preschool years, and revealed that the strength and consistency of preschoolers' knowledge of cognitive cuing and emotion was affected by whether cues were the same, or only similar to, parts of the earlier events. PMID- 9418228 TI - Children's understanding of epistemic conduct in self-deception and other false belief stories. AB - In self-deception persons accept false beliefs through a motivated disregard for countervailing evidence. Such epistemic misconduct renders them responsible for their own deception. It was hypothesized that children's understanding of this responsibility would be associated with an understanding of how evidence informs belief. In the study 4- to 9-year-old children's understanding of the relations between false belief, evidence, and epistemic responsibility was examined using stories involving self-deception, lying, and misleading appearances. Results indicated that younger children who understood false belief understood simpler types of deception, but that understanding self-deceivers' epistemic responsibility was limited to older children who understood the relevance of evidence to belief formation. PMID- 9418229 TI - Preschoolers' attributions of mental states in pretense. AB - When young children appear to recognize that someone else is engaging in make believe play, do they infer what the pretender is thinking? Are they aware that the pretender is thinking about a pretend scenario yet knows what the real situation is? Preschoolers ages 3-5 (N = 45) viewed scenes from the Barney & Friends television series depicting either make-believe or realistic actions. Children were questioned concerning the presence of pretense and the thoughts and beliefs of the TV characters. The children were also presented with false belief and appearance/reality theory of mind tasks. Children who identified when TV characters were engaging in pretend play did not necessarily infer the pretenders' thoughts and beliefs. Inferring pretenders' thoughts was related to performance on false belief and appearance/reality tasks, but simply recognizing pretense was not. These data support the view that children initially learn to recognize pretense from contextual cues and are able to infer pretenders' beliefs only with further development of metarepresentational ability. PMID- 9418230 TI - Preschoolers' attention to and memory for attachment-relevant information. AB - This study examined the relation between attachment quality in infancy and attention and memory at 3 1/2 years. Sixty-eight children participated in 2 attention tasks and 1 memory task. In the first attention task, children were shown several sets of drawings; each set depicted a different mother-child dyad engaged in positive, negative, and neutral interaction. Insecure/avoidant children looked away from the drawings more than the other children. In the second attention task, children were shown different sets of drawings; each set depicted a mother-child dyad engaged in positive interaction and an adult dyad expressing neutral affect. Insecure/avoidant and insecure/ambivalent children looked away from the mother-child drawings more than the secure children; when children did look at a drawing, insecure children were less likely than secure children to look at the mother-child drawing. In the memory task, children were read 6 stories in which a mother responds to her child's bid for help. In 2 stories the mother responds sensitively to her child, in 2 stories the mother rejects her child, and in 2 stories the mother provides an exaggerated response to her child. Secure children recalled the responsive stories better than insecure/avoidant children and the rejecting stories better than the insecure/ambivalent children. Findings are discussed in terms of the proposition from attachment theory that attachment experiences influence attention and memory processes. PMID- 9418231 TI - Children's representations of attachment relationships in family drawings. AB - Children's representational models of self and attachment figures were investigated in family drawings at age 8-9 in a high-risk, racially mixed sample. Drawings were scored using a series of specific signs and a group of theoretically derived, global rating scales. When specific signs were treated in a combined way (versus separately), they were significantly related to early attachment history in predicted ways. Similarly, specific rating scales were found to be significantly related to early relationship history. Analyses exploring the relative contributions of early attachment history and contemporary measures of child IQ, life stress, and emotional functioning revealed that even after contemporary influences were taken into account, attachment history made a significant contribution to the prediction of negative drawing outcome. Results were interpreted as supporting an organizational perspective on development where qualitative differences in early relationships are hypothesized to shape core representational models of the self and to exert an ongoing influence on later representational processes. PMID- 9418232 TI - Distinguishing multiple dimensions of conceptions of ability: implications for self-evaluation. AB - Three separate lines of research have suggested that conceptions of ability may play a key role in the development of self-evaluation. Each line has focused on a different dimension of conceptions of ability: conceptions of ability as uncontrollable, conceptions of ability as constant, and conceptions of ability as capacity. Unfortunately, there has been little attention to the convergences and divergences among the 3 dimensions. The present study examined this issue in 236 second- through fifth-grade children. Children indicated the extent to which they conceived of ability as uncontrollable, as constant, and as capacity. Two forms of self-evaluation (performance following failure and the extent to which self perceptions of competence converge with external indicators of competence) were investigated. In addition, cognitive competence was assessed. The near-zero correlations, 3-factor solution yielded by confirmatory factor analysis, variability in age-related differences, differential links to cognitive competence, and diverse forms of self-evaluation among the 3 dimensions suggested that the 3 are relatively distinct, and that they may play different roles in the development of self-evaluation. Moreover, the 3 dimensions appear to interact with one another to influence self-evaluation. PMID- 9418233 TI - Classroom peer acceptance, friendship, and victimization: distinct relational systems that contribute uniquely to children's school adjustment? AB - The proposition that relationships make differential (i.e., unique, redundant, contingent) contributions to adjustment was examined by investigating the linkages between children's participation in different types of peer relationships (i.e., friendship, peer acceptance, peer victimization) and their adjustment to school. Relationship measures were gathered for 5- to 6-year-old children (105 males, 95 females) twice during kindergarten (i.e., fall and spring) and were correlated with adjustment indicators at each time of assessment and used to predict changes in school adjustment over time. Examination of the relative associations between the relationship measures and children's adjustment revealed evidence of both unshared (i.e., unique) and shared (i.e., redundant) linkages, depending on the form of adjustment examined. These findings suggest that adjustment may be influenced by the diverse experiences (i.e., provisions) that children encounter in different forms of relationship, and that certain types of relationships may have greater or lesser adaptive significance depending on the adjustment outcome examined. PMID- 9418234 TI - Friendships, peer acceptance, and group membership: relations to academic achievement in middle school. AB - Two samples of sixth-grade students were followed over time to examine relations of number of reciprocated friendships, peer acceptance, and group membership to academic achievement. In both samples, group membership was the most consistent predictor of grades over time. In Study 2, prosocial behavior, antisocial behavior, and emotional distress were examined as processes that might explain these significant links between peer relationships and academic achievement. Results of longitudinal analyses support a conclusion that aspects of peer relationships are related to classroom achievement indirectly, by way of significant relations with prosocial behavior. Future research might benefit from more in-depth analyses of the functions of adolescent peer relationships and the processes by which they influence orientations toward social and academic competence at school. PMID- 9418235 TI - Adaptation of Bacillus subtilis to oxygen limitation. AB - Bacillus subtilis grows anaerobically by at least two different pathways, respiration using nitrate as an electron acceptor and fermentation in the absence of electron acceptors. Regulatory mechanisms have evolved allowing cells to shift to these metabolic capabilities in response to changes in oxygen availability. These include transcriptional activation of fnr upon oxygen limitation, a process requiring the ResD-ResE two-component signal transduction system that also regulates aerobic respiration. FNR then activates transcription of other anaerobically induced genes including the narGHJI operon which encodes a respiratory nitrate reductase. Genes involved in fermentative growth are controlled by an unidentified FNR-independent regulatory pathway. PMID- 9418236 TI - Identification of gene products from the Azotobacter vinelandii nifBfdxNnifOQ operon. AB - The Azotobacter vinelandii nifBfdxNnifOQ operon is required for synthesis of the nitrogenase iron-molybdenum cofactor. To further characterize the roles of its gene products, specific antibodies against NifB and NifO were generated, and the NifB, NifO and NifQ gene products were visualized and identified in nitrogen fixing A. vinelandii cell extracts by a combination of two-dimensional gel electrophoresis of radiolabelled extracts and immunological detection methods. The three proteins showed apparent pI and M(r) values similar to those expected from sequence data, except for NifO, which showed an apparent M(r) of ca. 23 kDa (vs. 16 kDa expected). PMID- 9418237 TI - Determinant role of E. coli RNase III in the decay of both specific and heterologous mRNAs. AB - A comparative analysis of mRNA decay was carried out in Escherichia coli using the wild-type and an isogenic RNase III deletion strain. We have studied the mRNA degradation from the Escherichia coli gene bolA, the Lactococcus lactis biovar diacetylactis citQRP operon and the Desulfovibrio vulgaris Hildenborough gene cyc. As seen by a dramatic stabilization of the specific mRNAs in the mutant strain, RNase III was crucial for the decay process of these three messages. Since RNase III, unlike RNase E, is not essential for bacterial viability we think that there is potential for using RNase III mutant strains to modulate gene expression. PMID- 9418238 TI - Role of the COOH-terminal pro-sequence of aqualysin I (a heat-stable serine protease) in its extracellular secretion by Thermus thermophilus. AB - Aqualysin I is a subtilisin-type serine protease secreted into the medium by Thermus aquaticus YT-1. Thermus thermophilus cells harboring a plasmid for the aqualysin I precursor secreted pro-aqualysin I with the C-terminal pro-sequence into the culture medium, and the precursor was then processed to the mature enzyme during the cultivation. However, the extracellular levels of aqualysin I in T. thermophilus cells harboring plasmids for deletion mutants as to the C terminal pro-sequence were about 10-20% in comparison with the level of wild type. Only the mature enzyme could be detected in the medium, while pro-aqualysin I with the C-terminal pro-sequence could not. These results suggest that the C terminal pro-sequence of aqualysin I plays an important role in the extracellular secretion of aqualysin I. PMID- 9418239 TI - Comparison of ViaB regions of Vi-positive organisms. AB - We cloned the vipR genes from Salmonella paratyphi C, S. dublin, and Citrobacter freundii strains and compared them with the S. typhi sequence to clarify the genetic relationship of the ViaB regions of Vi-positive organisms. ViaB regions were divided into two groups based on their sequences, the Salmonella and C. freundii groups. The vipR coding sequences of the Salmonella group were identical. Southern blot hybridization results using the full-length ViaB region as a probe support these findings. PMID- 9418240 TI - Characterization of a periplasmic peptidyl-prolyl cis-trans isomerase in Erwinia chrysanthemi. AB - The main determinant of the plant pathogen Erwinia chrysanthemi virulence is the production of extracellular enzymes, mainly pectate lyases. Adjacent to a pectate lyase encoding locus, we identified the gene rotA supposed to encode a folding catalyst. Overproduction of the protein and assay of activity using a synthetic substrate, confirmed that rotA encodes a periplasmic peptidyl-prolyl cis-trans isomerase. rotA disruption provokes no change in cell morphology, cell viability, growth rate or stability of the extracellular and periplasmic proteins. In addition, this mutation does not alter the activity of the pectate lyases, their stability in the periplasm during the transitory step of secretion or their recognition by the Out secretory system. rotA expression was followed using a rotA::uidA transcriptional fusion. Some environmental conditions, such as temperature variations and nitrogen starvation, modulate rotA expression. In contrast to the E. coli rotA gene, the E. chrysanthemi rotA possesses only one promoter and is not controlled by the CRP global regulator. PMID- 9418241 TI - Regulation of fumarase (fumB) gene expression in Escherichia coli in response to oxygen, iron and heme availability: role of the arcA, fur, and hemA gene products. AB - Three distinct fumarases, FumA, FumB and FumC, have been reported in Escherichia coli. While the fumA and fumC gene products are expressed under aerobic cell growth conditions, the FumB fumarase appears to be more abundant during anaerobic growth. To study the transcriptional regulation of the fumB gene, a fumB-lacZ operon fusion was constructed and analyzed in a single copy under a variety of cell culture conditions. Expression of fumB-lacZ was fourfold higher under anaerobic than aerobic growth conditions. This anaerobic response is modulated by the ArcA and Fnr proteins, which function independently as anaerobic activators of fumB gene expression. Cellular iron limitation in a fur mutant caused fumB lacZ expression to decrease sevenfold while cellular heme limitation decreased fumB gene expression twofold. In addition, fumB-lacZ expression was shown to vary depending on the DNA superhelicity. This study further delineates the regulation of the fumB gene in cell growth. PMID- 9418242 TI - Molecular cloning and sequence analysis of the lysR gene from the extremely thermophilic eubacterium, Thermus thermophilus HB27. AB - We have isolated a lysine-auxotrophic and kanamycin-resistant mutant from an extreme thermophile, Thermus thermophilus HB27. This mutant showed the lysA- or lysR- genotype since it could not grow on the minimal plate which contained diaminopimelic acid. Sequence analysis of the clones which could rescue the Lys- mutant indicated the lysR gene. The lysR gene overlapped with the rimK gene for the modification enzyme of ribosomal protein S6. In the Lys- mutant, the lysR gene was disrupted and the C-terminus region of the RimK protein was different from that of the wild-type, which contributed to the Lys- and kanamycin-resistant phenotype. The deduced amino acid sequence of the lysR gene showed 20.9% identity with the LysR protein of Escherichia coli. The percentage of use of cytosine or guanine in the third letter of the codons in the lysR gene was only 67.4%. We also determined that the argC gene encoding N-acetyl-gamma-glutamyl phosphate reductase and the argB gene encoding acetylglutamate kinase were located immediately upstream of the lysR gene. PMID- 9418243 TI - Single strand conformation polymorphism analysis of PCR-tDNA fingerprinting to address the identification of Bacillus species. AB - The suitability of tDNA-PCR fingerprinting to identify species of the genus Bacillus was tested on 75 strains. Strains belonging to the same species or the same phylogenetic cluster were correctly grouped. Among B. stearothermophilus strains, different pattern types were found. This could be due to the unclear taxonomic situation of these strains, rather than to a failure of the tDNA-PCR. Single strand conformation polymorphism (SSCP) of the PCR products allowed species discrimination within the 'B. subtilis group', but not within the 'B. cereus group'. The tDNA-PCR, alone or coupled with SSCP analysis, is useful to address Bacillus species identification, particularly for those species which are not phylogenetically tightly clustered. PMID- 9418244 TI - Purification and characterization of 2,4-dichlorophenol hydroxylase isolated from a bacterium of the alpha-2 subgroup of the Proteobacteria. AB - 2,4-Dichlorophenol hydroxylase (EC 1.14.13.20) was purified to apparent homogeneity from the bacterial strain S1, a member of the alpha-2 subgroup of the Proteobacteria. The molecular masses of the native enzyme and the subunit were determined to be 256 and 64 kDa, respectively, suggesting a homotetrameric structure. The enzyme converted 2,4-dichlorophenol to 3,5-dichlorocatechol. The apparent K(m) values for 2,4-dichlorophenol, NADPH and NADH were 3, 240 and 420 microM, respectively. The enzyme hydroxylated a broad range of halogenated phenols. 3-Chloro-, 2,6-dichloro- and 2,4,6-trichlorophenol acted as 'non substrate' effectors. The N-terminal sequence revealed 72% identity with the amino acid sequence deduced from the pJP4-encoded tfdB gene. PMID- 9418245 TI - An E. coli B mutation, rpoB5081, that prevents growth of phage T4 strains defective in host DNA degradation. AB - An E. coli B Tab strain, EM121, was isolated that restricts T4 denA (DNA endonuclease II) mutants at 37 degrees C and above, but is permissive for wild type T4 at all temperatures examined. At 42 degrees C, other mutants affected in nucleic acid metabolism (T4 dexA, regA and uvsW strains) are also restricted. Genetic analysis revealed that one mutation (rpoB5081) in the RNA polymerase beta subunit gene is sufficient for restricting all denA mutants. rpoB5081, together with a second linked mutation, is also required for restricting the other T4 mutants, rpoB5081 (P806S), previously shown to increase transcription termination in E. coli K-12, causes delayed synthesis of T4 late proteins and reduced DNA synthesis in denA infections. Thus, T4 DNA synthesis and gene expression are impaired by the rpoB5081 beta subunit when degradation of host DNA is reduced. Because the restricted T4 mutants are not readily distinguished from wild-type phage under typical plating conditions, EM121 is an important host for screening and mapping T4 denA mutations. PMID- 9418246 TI - Cloning and characterization of the lytB gene of Campylobacter jejuni. AB - LytB of Escherichia coli is an essential gene involved in penicillin tolerance and the stringent response. The lytB gene of Campylobacter jejuni was cloned and characterized. It could complement a temperature-sensitive E. coli lytB mutant. The C. jejuni lytB gene encodes a protein of 277 amino acids that has 34, 36 and 40% amino acid identity with the LytB proteins of E. coli, Haemophilus influenzae, and Synechocystis sp. PCC6803, respectively. The lytB gene is situated between the aroA gene and a gene that encodes ribosomal protein S1. PMID- 9418247 TI - Enzyme-linked immunosorbent assay for detection of Aeromonas hydrophila serogroup O:19. AB - An enzyme-linked immunosorbent assay has been developed for the detection of Aeromonas hydrophila serogroup O:19 isolated from epizootics in eels. The enzyme linked immunosorbent assay specificity was confirmed after testing A. hydrophila O:19 and non-O:19 strains from different origins, as well as other Aeromonas species and other fish pathogens such as Vibrio vulnificus biotype 2, V. furnisii, V. damsela, Yersinia ruckerii and Edwardsiella tarda. The detection limits for A. hydrophila O:19 cells were around 10(4)-10(5) cells/well. Artificially infected eels were analyzed and the immunodetection was confirmed by cultural methods. With this methodology A. hydrophila O:19 was successfully detected in infected eels and water samples. We described two subgroups within the serogroup O:19 (Guinee and Jansen system), one of them presents a 50 kDa outer membrane protein as a strong thermostable antigen which is not present in the other group. PMID- 9418248 TI - Involvement of MAP-kinases and -phosphatases in uptake and intracellular replication of Listeria monocytogenes in J774 macrophage cells. AB - In this study we show that protein tyrosine kinases and also protein tyrosine phosphatases are involved in the uptake of Listeria monocytogenes by J774 macrophages to a different extent than in the uptake of inert latex beads. In addition, protein tyrosine kinases are necessary for the intracellular growth and survival of L. monocytogenes. The expression of the MAP kinase phosphatase MKP-1, a protein tyrosine phosphatase, is induced upon infection, and phagocytosis of L. monocytogenes by J774 cells overexpressing the MKP-1 protein is reduced compared to control cells. The decreased phagocytosis of L. monocytogenes as a result of the MKP-1 overexpression in J774 macrophages suggests that the activation of the MAP kinase(s) ERK-1 and/or ERK-2 is an essential requirement for the uptake of L. monocytogenes by J774 macrophages. PMID- 9418249 TI - Functional analysis of a conserved aspartate D218 in Cephalosporium acremonium isopenicillin N synthase. AB - Isopenicillin N synthase (IPNS) is instrumental in the catalytic conversion of a tripeptide precursor delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine to a bioactive intermediate isopenicillin N in the beta-lactam antibiotic biosynthetic pathway. It has recently been shown that this reaction is dependent on a conserved aspartate, D214, in a bacterial Streptomyces jumonjinensis IPNS. Thus, this study was carried out to provide the experimental evidence for the involvement of a similarly conserved aspartate residue, D218, in a fungal Cephalosporium acremonium IPNS (cIPNS). Initially, alteration of the aspartate residue to generate the mutant D218L cIPNS protein was achieved by site-directed mutagenesis. Subsequent enzyme assays indicated that the catalytic property of the mutant protein was lost, attesting to the need for the corresponding conserved aspartate to maintain IPNS functionality. It is also evident from the observed results that site-directed mutagenesis of this particular aspartate residue in cIPNS can affect its solubility. It is therefore important to take these potential changes into consideration when site-directed mutant proteins are analysed for catalytic function. PMID- 9418250 TI - Synthesis of poly(3-hydroxyalkanoates) in Escherichia coli expressing the PHA synthase gene phaC2 from Pseudomonas aeruginosa: comparison of PhaC1 and PhaC2. AB - In order to obtain functional expression of PHA synthase gene phaC2 from Pseudomonas aeruginosa in Escherichia coli, the coding region of phaC2 was subcloned, including the ribosomal binding site, into pBluescript SK- collinear to the lac promoter. This plasmid pBHR71-C2 enabled functional expression of phaC2 in E. coli LS1298 (fadB) under lac promoter control, leading to PHA accumulation, when grown in LB medium containing 0.5% (w/v) of various fatty acids (C8-C14). The strongest accumulation of PHA was observed, when dodecanoate was provided as carbon source, and PHA contributed to 15% of cell dry weight, which was composed of 35 mol% 3-hydroxydodecanoate, 60 mol% 3-hydroxydecanoate and 5 mol% 3-hydroxyoctanoate. Plasmid pBHR78, which contained both genes phaC1 and phaC2 from P. aeruginosa under lac promoter control in pBluescript SK- led in E. coli LS1298 to PHA accumulation, which contributed to 13% of cell dry weight, when cells were grown on decanoate. Only slight differences in PHA composition compared with either PhaC1 or PhaC2 were obtained. The weight average molecular masses of PHA purified from decanoate-grown cells of E. coli LS1298 expressing PhaC1 or PhaC2 alone or both PHA synthases, were 106 x 10(3), 70 x 10(3) or 67 x 10(3), respectively. This study clearly demonstrated that both PHA synthases from P. aeruginosa exhibit very similar properties resulting in similar extent of PHA accumulation, similar composition and molecular mass, when expressed in E. coli and that fatty acid beta-oxidation provides substrates for both PHA synthases. PMID- 9418251 TI - Listeria monocytogenes-infected human umbilical vein endothelial cells: internalin-independent invasion, intracellular growth, movement, and host cell responses. AB - The interaction of Listeria monocytogenes with human umbilical vein endothelial cells was studied. We show that L. monocytogenes invades human umbilical vein endothelial cells independently of internalin A, internalin B, internalin C, and ActA. L. monocytogenes replicates efficiently inside the cells and moves intracellularly by the induction of actin polymerization. We further show that L. monocytogenes-infection of human umbilical vein endothelial cells induces interleukin-6 and interleukin-8 expression during the first 6 h of infection. The expression of MCP-1 and the adhesion molecules VCAM-1 and ICAM-1 was not altered under the experimental conditions used here. PMID- 9418252 TI - Inhibition of metallo-beta-lactamases by a series of thiol ester derivatives of mercaptophenylacetic acid. AB - A series of mercaptophenylacetic acid thiol esters bearing a phenyl substituent adjacent to the carboxylic acid function has been shown to be inhibitors of metallo-beta-lactamases. The inhibition of the Bacteroides fragilis CfiA and Bacillus cereus II metallo-beta-lactamases was Zn2- dependent, greater inhibition being observed at 1 microM ZnSO4 than at 100 microM ZnSO4. Despite this Zn2+ dependency, isothermal titration calorimetry studies illustrated that representative compounds had no detectable affinity for Zn2+ (K > 1 mM). This indicates that their mode of inhibition was not by chelation of the active site Zn2+. Greatest potency was observed against the Stenotrophomonas maltophilia L1 metallo-beta-lactamase with I50 values of between < 1.95 microM and 6 microM and SB-217843 exhibited a similar level of inhibition of this enzyme at 1 and 100 microM Zn2+ (I50 values 5 and 6 microM, respectively). Inhibition of B. cereus II metallo-beta-lactamase by SB-218018 and SB-217782 was competitive with Ki values of 185 microM and 1500 microM, respectively. Therefore, these compounds are specific inhibitors of metallo-beta-lactamases and provide further probes of the active sites of these enzymes. PMID- 9418253 TI - Characterisation of integrons and antibiotic resistance genes in Danish multiresistant Salmonella enterica Typhimurium DT104. AB - The presence and genetic content of integrons was investigated in eight Salmonella enterica Typhimurium DT104 isolates from different pig herds in Denmark. Two different integrons were identified using PCR and sequencing. Each of the integrons carried a single resistance cassette in addition to the sul1 and qacE delta 1 genes characteristic of integrons. The first integron encoded the ant (3")-Ia gene that specified resistance to spectinomycin and streptomycin. The second contained the pse-1 beta-lactamase gene. All the multiresistant strains contained both integrons. The presence of these two integrons did not account for the total phenotypic resistance of all the isolates and does not exclude the presence of other mobile DNA elements. PMID- 9418254 TI - Lipopolysaccharide from Burkholderia vietnamiensis strain LMG 6999 contains two polymers identical to those present in the reference strain for Burkholderia cepacia serogroup O4. AB - Lipopolysaccharide was isolated from strain LMG 6999 of Burkholderia vietnamiensis. Degradative and NMR spectroscopic studies established the presence of two polymeric fractions based on the following trisaccharide repeating units: I:-->3)-alpha-D-Galp-(1-->3)-beta-D-Galp-(1-->3)-beta-D-GalpNAc- (1-->; II:-->3) alpha-D-GalpNAc-(1-->3)-beta-D-GalpNAc-(1-->4)- alpha-L-Rhap-(1-->. The same polymers have previously been found together in lipopolysaccharide from the reference strain for Burkholderia cepacia serogroup O4 and, individually, in those from B. cepacia serogroups C (I) and A (II). PMID- 9418256 TI - Amplification of bacitracin transporter genes in the bacitracin producing Bacillus licheniformis. AB - We have amplified the previously cloned and sequenced genes of the bacitracin exporter (bcr), a member of the ATP-binding transport protein family, within the chromosome of the bacitracin producing Bacillus licheniformis. Amplification of the transporter genes was followed by greatly increased bacitracin resistance. Antibiotic production was enhanced at a low level of bcr genes amplification. An enlarged increase in the copy number of the bcr genes negatively affects the overall growth of bacteria. PMID- 9418255 TI - Streptomyces coelicolor DNA homologous with acyltransferase domains of type I polyketide synthase gene complex. AB - An acyltransferase-homologous DNA fragment was amplified in a PCR reaction on a cosmid DNA template from the genomic DNA library of the soil bacterium Streptomyces coelicolor A3(2). The putative amino acid sequence of the fragment resembles acyl-CoA:ACP acyltransferase domains from several bacterial enzymatic complexes of polyketide synthase. There is a high similarity with acyltransferase domains from so-called type I polyketide synthases. Such synthases catalyze production of the aglycone portion of macrolides and polyethers that are important as antibiotics or immunosuppressants. The amplified fragment is considered to be a part of a larger gene complex. PMID- 9418257 TI - Comparison of the abilities of proteins from Bartonella bacilliformis and Bartonella henselae to deform red cell membranes and to bind to red cell ghost proteins. AB - Infections in humans by Bartonella bacilliformis, but not Bartonella henselae, are characterized by invasion of red cells. Supernatants of culture medium from B. bacilliformis and B. henselae each contain a protein which causes invagination of membranes of human red cells and formation of intracellular vacuoles. These two proteins are very similar in molecular mass, heat stability and mechanism of action. B. henselae does not bind to human red cells, but human red cell ghost membrane proteins were recognized by both bacteria, five by B. bacilliformis and the same five, and one additional protein by B. henselae. Two of these proteins had molecular masses consistent with actin and spectrin. Actin binds to five electroblotted outer membrane proteins from B. henselae and four of these proteins are retained on an actin-Sepharose column. PMID- 9418258 TI - Genetic variation and interspecific hybridization among natural populations of zoysiagrasses detected by RFLP analyses of chloroplast and nuclear DNA. AB - Genetic variations among 17 accessions of zoysiagrasses collected from natural populations in Japan were investigated by RFLP analyses of chloroplast DNA (cpDNA) and nuclear DNA. These accessions were classified into five species based on morphological characteristics: Zoysia japonica, Z. matrella, Z. tenuifolia, Z. sinica, and Z. macrostachya. On the basis of eight kinds of RFLPs in cpDNAs detected across accessions, six chloroplast genome types (types A-F) were identified. Although type-A cpDNA was shared by five accessions of japonica and four accessions of matrella, derivative cpDNAs of type A, which each arose by a mutation, were identified in one accession of japonica (type B) and in two accessions of matrella (type C). One accession of japonica which showed spikelets similar to those of shapes macrostachya, contained type-F cpDNA as did sinica and macrostachya. The two accessions of tenuifolia each showed a specific cpDNA type, i.e. types D and E. Genetic relationships among the 17 accessions were investigated by the RFLP analyses of nuclear DNA with 20 genomic and gene probes. A dendrogram constructed with genetic distances calculated from the RFLP patterns indicated four major groups among them. Six accessions of japonica comprised one group, whereas the one accession of japonica possessing the type-F cpDNA was clustered with macrostachya and sinica. Four accessions of matrella with type A cpDNA constituted another group in the dendrogram, showing a closer relationship to the japonica accessions than to the other two accessions of matrella. The remaining two accessions of matrella and tenuifolia accessions were grouped together. These data indicate that zoysiagrasses distributed in Japan harbor highly genetic variations, and that interspecific hybridization has occurred in natural populations. PMID- 9418259 TI - Detection of the Sec-1 locus of rye by a PCR-based method. AB - The structural genes for the omega-secalins of rye (Secale cereale) are located in the Sec-1 locus on the short arm of rye chromosome 1R. We applied PCR (polymerase chain reaction) to detect the Sec-1 locus in a wheat genomic background. A primer set we designed based on a published sequence of a omega secalin gene amplified not only the omega-secalin sequence, but also a putative omega-gliadin sequence. We determined partial sequences of both PCR-amplified fragments and designed different primers for the specific amplification of the omega-secalin sequence. One of the new primer sets amplified DNA fragments only in rye and wheat lines carrying chromosome 1R or telosome 1RS; no amplification occurred in either euploid wheats or 1RS deletion lines. This PCR-based method would provide efficient screening for the Sec-1 locus in progeny of wheat lines carrying chromosome 1R. PMID- 9418260 TI - A phylogenetic analysis of three group I introns found in the nuclear small subunit ribosomal RNA gene of the ballistoconidiogenous anamorphic yeast-like fungus Tilletiopsis flava. AB - There are three group I introns in the nuclear small subunit ribosomal RNA gene (SSU rDNA) of the ballistoconidiogenous anamorphic yeast-like fungus Tilletiopsis flava JCM 5186. The size of these sequences were 325 nt (position 516), 335 nt (position 1199) and 437 nt (position 1506), respectively. The introns at position 516 (T.flav516) and position 1199 (T.flav1199) belonged to subgroup IB3, and that of position 1506 (T.flav1506) belonged to subgroup IC1. The results of comparison with other group I introns found in SSU rDNA of eucaryotes showed that the positions 516 and 1199 were common positions to IB3 group I introns of fungi and green algae, and that positions 943, 1506 and 1512 were those to IC1 group I introns of fungi, and green and red algae. It is indicated that the insertion position of introns have close relationship with the nature of the subgroup to which they belonged. For phylogenetic analysis, we employed 9 IB3 introns, in which 7 were at position 516 and 2 were at position 1199, and 25 IC1 introns. The maximum likelihood tree based on the conserved region alignment showed that group I introns of subgroup IB3 were phylogenetically distant from those of subgroup IC1. T.flav516 (basidiomycete) constituted a subcluster with R.dacr516 (basidiomycete) and M.albo516 (ascomycete). T. flav1199 was located at the closer position of C.chlo1199 (green alga) than other IB3 introns at position 516. T.flav1506 was located at the subcluster, which was constituted by the 1506 introns found in SSU rDNA of fungi (B.yama1506, P.cari1506, and P.inou1506) and those of green algae (C.elli1506, C.mira1506, G.spir1506, and M.sacl1506) with IC1 introns at the position 1512 (D.parv1512 and C.sacc1512). The analysis of flanking regions showed that both 5' and 3' flanking sequences were well conserved in each insertion site, and indicated that the ancestors of the intron at different site had been inherited from the different origin. Therefore, the two IB3 introns found positions 516 and 1199, T.flav516 and T.flav1199, were supposed to have the independent ancestors. Our results supported the theory of the diversity of group I introns that group I introns had been transferred horizontally to the distinct insertion site, and were inherited and diverged vertically. PMID- 9418261 TI - Localization of the genes for major ribosomal RNA on chromosomes of the house musk shrew, Suncus murinus, at meiotic and mitotic cells by fluorescence in situ hybridization and silver staining. AB - The genes for major ribosomal RNA were localized on chromosomes 5pter-p15, 9q64 qter, and 13q38-qter of the house musk shrew, Suncus murinus (Insectivora, Soricidae) by silver staining of mitotic metaphase and meiotic pachytene spreads and fluorescence in situ hybridization using the human 28S-RNA genes as a probe to mitotic metaphase spreads. The data presented indicate a correlation between sites of in situ hybridization and silver staining. The finding of nuclear materials in mitosis was in a good agreement with observation in meiosis: same chromosomes carried active NORs in both meiotic and mitotic cells. PMID- 9418262 TI - Cloning and characterization of a mariner-like element in the silkworm, Bombyx mori. AB - We have identified a mariner-like element (MLE) in the genome of the silkworm, Bombyx mori. The cloned MLE, desiqnated as BmMLE, contained an ORF that was interrupted by insertions, stop codons, and frameshifts. The complete ORF sequence was estimated by sequencing the PCR products. The hypothetical protein coded by this ORF showed 34% homology to the Mos1 element of Drosophila mauritiana, and is phylogenetically very close to MLE of Hyalophora cecropia. BmMLE is present in 80 to 100 copies in the B. mori genome. Many BmMLEs with different lengths were identified by Southern blot analysis and LA PCR of the genomic DNA. The distribution of these BmMLEs differed in geographic races. PMID- 9418263 TI - Extensive interbreeding occurred among multiple matriarchal ancestors during the domestication of dogs: evidence from inter- and intraspecies polymorphisms in the D-loop region of mitochondrial DNA between dogs and wolves. AB - To test the hypothesis that the domestic dogs are derived from several different ancestral gray wolf populations, we compared the sequence of the displacement (D) loop region of the mitochondrial DNA (mtDNA) from 24 breeds of domestic dog (34 individual dogs) and 3 subspecies of gray wolf (Canis lupus lupus, C.l. pallipes and C.l. chanco; 19 individuals). The intraspecific sequence variations within domestic dogs (0.00-3.19%) and within wolves (0.00-2.88%) were comparable to the interspecific variations between domestic dogs and wolves (0.30-3.35%). A repetitive sequence with repeat units (TACACGTA/GCG) that causes the size variation in the D-loop region was also found in both dogs and wolves. However, no nucleotide substitutions or repetitive arrays were specific for domestic dogs or for wolves. These results showed that there is a close genetic relationship between dogs and wolves. Two major clades appeared in the phylogenetic trees constructed by neighbor-joining and by the maximum parsimony method; one clade containing Chinese wolf (C.l. chanco) showed extensive variations while the other showed only slight variation. This showed that there were two major genetic components both in domestic dogs and in wolves. However, neither clades nor haplotypes specific for any dog breed were observed, whereas subspecies-specific clades were found in Asiatic wolves. These results suggested that the extant breeds of domestic dogs have maintained a large degree of mtDNA polymorphisms introduced from their ancestral wolf populations, and that extensive interbreedings had occurred among multiple matriarchal origins. PMID- 9418264 TI - Developmental constraints in the Drosophila wing. AB - Selection experiments for shortening the four longitudinal veins in a wild population of Drosophila melanogaster have been performed to evaluate how a local change is integrated in the wing development. Our results show that, though many units of selection seem to exist within a given organ, these are strongly constrained within the developmental programme, in such a way that only some predictable forms are expected. The results are discussed in terms of the 'Entelechia' model proposed by Garcia-Bellido in which the intercalarity of positional values promoted by 'martial' genes in a given organ is the driving force for controlled cell proliferation. PMID- 9418265 TI - Developmental constraints and wing shape variation in natural populations of Drosophila melanogaster. AB - The body sizes and shapes of poikilothermic animals generally show clinal variation with latitude. Among the environmental factors responsible for the cline, temperature seems to be the most probable candidate. In the present work we analysed natural populations of Drosophila melanogaster collected at different geographical localities to determine whether the same selective forces acting on wing development in the laboratory are also at work in the wild. We show that the temperature selection acting on wing development in the laboratory is only one of the selective forces operating in the wild. The size differences between natural populations seem to depend exclusively on cell number whereas they depend on cell area in the laboratory. The two wing compartments behave as distinct units of selection subjected to different genetic control, confirming our previous observations on laboratory populations. In addition, subunits of development defined as regions of cell proliferation centres restricted within longitudinal veins can, in turn, be considered as subunits of selection. Their interaction during development and continuous natural selection around an optimum could explain the high wing shape stability generally found in natural populations. PMID- 9418266 TI - Correlations among size-related traits are affected by chromosome inversions in an adaptive polymorphism in Drosophila buzzatii. AB - Genetic variation in correlations among size-related traits of head, thorax and wings was examined in Drosophila buzzatii, by comparing the correlation pattern of the phenotypic correlation matrix (CP-Rp) between inversion karyotypes of the second chromosome. CP-Rp differed between some karyotypes in a natural population. CP-Rp in homokaryotypic classes of wild-reared flies, but not in heterokaryotypes, differed from the whole population represented by laboratory reared flies. Similarity in CP-Rp was highly significant for a same homokaryotype in two populations. In one of them, the chromosome is polymorphic for four inversions. In the other population, one of the inversions is almost fixed. CP-Rp was significantly similar between these populations, illustrating that similarity of CP-Rp may even occur between populations which have greatly diverged in frequencies of some genotypes affecting correlation patterns. It is suggested that chromosomal inversions are factors affecting genetic correlations among traits known to be phenotypically correlated with fitness components. PMID- 9418267 TI - More efficient breeding systems for controlling inbreeding and effective size in animal populations. AB - A selection scheme and a mating scheme are proposed to control the inbreeding and genetic drift in conserved or control animal populations with different numbers of males and females. Recurrence equations for the inbreeding coefficient and formulae for effective size are derived for autosomal loci, sex-linked loci with males being heterogametic and sex-linked loci with females being heterogametic under each of four breeding systems. It is shown that both the selection scheme and the mating scheme proposed in this paper could increase the effective size and decrease inbreeding in any generation compared with the classical selection and mating schemes. Among the four breeding systems considered, the most efficient one could increase the effective size by as much as 19 per cent for autosomal loci and 50 per cent for sex-linked loci in comparison with the classical breeding system usually utilized in conserved or control populations. PMID- 9418268 TI - Nonrandom partition of mitochondria in heteroplasmic Drosophila. AB - In order to understand the status of heteroplasmy and its evolution within the Drosophila melanogaster subgroup, cytoplasm microinjections between eggs were performed involving three lineages of Drosophila simulans, carrying the siI, siII or siIII mtDNA type, respectively, and two strains of Drosophila mauritiana carrying the maI or maII mtDNA type. Progeny of eggs from all combinations of injection were analysed. The maII or siI molecules, when provided by the donor, were never detected in the offspring of the hosts, whatever the host's mitochondrial type. Heteroplasmic flies were detected when siII, siIII or maI mitochondria were injected into any of the other cytoplasms. In the majority of cases the percentage of foreign mtDNA increased over generations, leading to a complete replacement of the endogenous mtDNA. In most cases, siII was prevalent. The stochastic processes involved in the evolution of heteroplasmic states are strongly affected by selective values of the different mtDNA types, with a clear hierarchy among them: siII has the most advantage, then siIII and maI, and finally siI and maII. In the siII/maII heteroplasmy case, the loss of maII was more rapid at a high temperature. PMID- 9418269 TI - X-ray microscopy and imaging of Escherichia coli, LPS and DNA. AB - Ultrastructural examination by transmission and scanning electron microscopy involves a series of specialized preparation steps which may introduce artefacts in the micrographs. X-ray microscopy can take instant images of specimens but is mostly restricted to a few synchrotron X-ray sources. We have utilized a bench top nanosecond laser-plasma to produce a single-shot source of nanosecond X-rays tuned for maximum contrast with carbon-rich material. To examine the ultrastructure by absorption profiles, we utilized a laser-produced plasma generated by a single-shot laser (1.06 microns wavelength, 5 x 10(12) W cm-2 intensity) focused on to a silicon target as an X-ray source for high-resolution X-ray microscopy. This approach eliminates the specimen preparation steps. Whole hydrated cells of Escherichia coli and purified preparations of lipopolysaccharide (LPS) and chromosomal DNA (cDNA) were streaked onto poly(methyl methacrylate) (PMMA)-coated grids (resist). This resist was exposed to X-rays under vacuum at a distance of 2.5 cm from the target disc. The silicon plasma produced by a 10-ns burst of laser energy (at 20J) radiates strong emission lines in the region of 300 eV. The X-rays penetrate the sample and their absorption profile is transferred on to the resist where PMMA acts as a negative to generate an image. By atomic force microscopy imaging of this photoresist we have visualized layers around cells of E.coli, darker areas inside the cell probably corresponding to cDNA, and preliminary images of LPS and DNA molecules. This technique has resolution at the 100 A level, produces images similar to the space-filling models of macromolecules and may be of great value in the study of the ultrastructure of hydrated live biological specimens. PMID- 9418270 TI - Transmission X-ray microscopy of intact hydrated PtK2 cells during the cell cycle. AB - Transmission X-ray microscopy makes it possible to investigate biological specimens, i.e. cells and organelles, in their natural wet environment. The main processes determining the contrast in X-ray microscopy are photoelectric absorption and phase shift. X-ray microscopic experiments can therefore be carried out in both amplitude and phase contrast. The Gottingen X-ray microscope at the BESSY storage ring in Berlin is described. PtK2 cells were examined during different stages of the cell cycle. All major constituents of the mitotic apparatus, e.g. chromosomes, centromeres, microtubules and centrosomes, could be visualized, as well as the main structural compartments and organelles of the interphase cell, e.g. nuclear membrane, interphase chromatin, nucleolus and cytoplasmic mitochondria, as well as parts of the cytoskeletal apparatus. In this way new information can be obtained with regard to the ultrastructure of the constituents of intact and unstained cells at a resolution which bridges the gap between light microscopy and electron microscopy. The prospects for the future application of transmission X-ray microscopy in biomedical research are discussed. PMID- 9418271 TI - Reproducibility and accuracy of interactive segmentation procedures for image analysis in cytology. AB - The segmentation of nuclear images is a crucial step in the development of procedures using image analysis for the cytological diagnosis of cancer. The purpose of this study is to evaluate the reproducibility and accuracy of several interactive segmentation methods which can be used in this context. Four methods were studied: a thresholding-based method enabling selection of intensity histogram contrast and brightness, manual tracing with a stylus, and arc- and ellipse-fitting routines. Features of nuclear size and shape were derived from nuclei segmented on repeated occasions by several individuals. Variance component models provided a statistical framework for evaluating the intraobserver and interobserver variability of these measurements in terms of their intraclass correlation coefficients. Of the methods tested, the arc-fitting segmentation method gave the most reproducible results, and thresholding the least. Reproducibility was generally very high both between individuals and for repeated segmentations by a single individual. Accuracies of area measurements for the various methods, as determined with respect to point counting, paralleled the reproducibilities of the methods. Sample size requirements were observed to be more dependent on the biological variability of the tissue sampled than on the particular segmentation method or on the number of individuals performing segmentation. PMID- 9418272 TI - Reconstruction of optical pathlength distributions from images obtained by a wide field differential interference contrast microscope. AB - An image processing algorithm is presented to reconstruct optical pathlength distributions from images of nonabsorbing weak phase objects, obtained by a differential interference contrast (DIC) microscope, equipped with a charge coupled device camera. The method is demonstrated on DIC images of transparent latex spheres and unstained bovine spermatozoa. The images were obtained with a wide-field DIC microscope, using monochromatic light. After image acquisition, the measured intensities were converted to pathlength differences. Filtering in the Fourier domain was applied to correct for the typical shadow-cast effect of DIC images. The filter was constructed using the lateral shift introduced in the microscope, and parameters describing the spectral distribution of the signal-to noise ratio. By varying these parameters and looking at the resulting images, an appropriate setting for the filter parameters was found. In the reconstructed image each grey value represents the optical pathlength at that particular location, enabling quantitative analysis of object parameters using standard image processing techniques. The advantage of using interferometric techniques is that measurements can be done on transparent objects, without staining, enabling observations on living cells. Quantitative use of images obtained by a wide-field DIC microscope becomes possible with this technique, using relatively simple means. PMID- 9418273 TI - Evaluation of 26Mg stable isotope as an in vivo tracer of magnesium transport for SIMS ion microscopy imaging studies. AB - Isotopic detection with high sensitivity, one of the most important features of ion microscopy, allows the in vivo application of stable isotopes as tracers for unravelling smaller tissue structures implicated with transport capabilities. The evaluation of the mass interferences associated with a particular mass of secondary ion signals is a necessity for tracer studies with stable isotopes. We have tested the feasibility of 26Mg stable isotope as a tracer of magnesium transport in the killifish. The fish were given a single intraperitoneal injection of 3 mumol 26MgCl2.6H2O (99.5% 26Mg enrichment), and the renal distribution of 26Mg was examined in frozen freeze-dried cryosections with ion microscopy. High-mass resolution analyses were performed to evaluate the purity of positive secondary ion signals of the nominal masses 24, 25 and 26 in order to assess the purity of 24Mg, 25Mg and 26Mg signals, respectively, in kidneys of control and 26Mg-injected fish. In kidneys of control fish, the purities of 24Mg, 25Mg and 26Mg signals were approximately 97%, 82% and 90%, respectively. In fish that were injected with 26Mg stable isotope, an enhancement of 26Mg+ secondary ion signals was observed with signal purity reaching 95%. These observations indicate that 26Mg can be used successfully as a tracer of magnesium transport in animal models. To uncover the distribution of tracer 26Mg from the naturally abundant background of this isotope, a pixel-by-pixel digital subtraction is applied to the raw ion microscopy mass 26 image. PMID- 9418274 TI - Hedonic consequences of social comparison: a contrast of happy and unhappy people. AB - Two studies tested the hypothesis that self-rated unhappy individuals would be more sensitive to social comparison information than would happy ones. Study 1 showed that whereas unhappy students' affect and self-assessments were heavily affected by a peer who solved anagrams either faster or slower, happy students' responses were affected by the presence of a slower peer only. These between group differences proved to be largely independent of 2 factors associated with happiness, i.e., self-esteem and optimism. Study 2 showed that whereas the unhappy group's responses to feedback about their own teaching performance were heavily influenced by a peer who performed even better or even worse, happy students' responses again were moderated only by information about inferior peer performance. Implications for our appreciation of the link between cognitive processes and "hedonic" consequences are discussed. PMID- 9418275 TI - Social categorization and perceptual judgment of size: when perception is social. AB - Social knowledge may affect not only people's thoughts and judgments but also their actual perceptions of physical magnitude. The physical magnitude of a stimulus is perceived in a relative way, comparing the magnitude of the target with surrounding context stimuli. Because similar objects invite comparison processes more easily than dissimilar objects ("similarity breeds comparability"), social knowledge can affect judgments of physical magnitude by determining what is perceived as (dis) similar. In Experiment 1, the authors show that social categorizations that are based on physical cues (e.g., gender) may affect the magnitude of perceptual contrast effects (the Ebbinghaus illusion). More important, in Experiment 2, the influence of social categorizations that have no physical bases is shown to affect the magnitude of perceptual contrast effects. Implications of these findings for theories of social knowledge effects are discussed. PMID- 9418276 TI - External intergroup threat as an antecedent to perceptions of in-group and out group homogeneity. AB - The present research examined the relationship between external intergroup threat and perceptions of group variability. The first study found that when Texas A&M University students worked on a task in which students from a rival university were allegedly biased against them, they perceived more intragroup similarities versus differences than in an out-group benevolent condition and a control condition, and they also perceived the self as more similar to the in-group and more different from the out-group. These results were replicated in a second study, which used the same methodology except that the benevolent condition was excluded. The findings are discussed in terms of different reactions that individuals have to internal and external intergroup threat. PMID- 9418277 TI - Hot years and serious and deadly assault: empirical tests of the heat hypothesis. AB - Two archival studies examined the relation between year-to-year shifts in temperature and violent and property crime rates in the United States. Study 1 examined the relation between annual average temperature and crime rate in the years 1950-1995. As expected, a positive relation between temperature and serious and deadly assault was observed, even after time series, linear year, poverty, and population age effects were statistically controlled. Property crime was unrelated to annual average temperature. Study 2 examined the relation between the average number of hot days (> or = 90 degrees F) and the size of the usual summer increase in violence for the years 1950-1995. As expected, a positive relation was observed between number of hot days and magnitude of the summer effect, even after time series and linear year effects were statistically controlled. For property crime, the summer effect was unrelated to number of hot days. PMID- 9418278 TI - Higher-order factors of the Big Five. AB - Estimated factor correlations from 14 studies supporting the 5 factor, Big Five model of personality trait organization--5 studies based on children and adolescents, 9 on adults--were factor analyzed. Two higher-order factors were clearly evident in all studies. One was principally related to the Big Five trait dimensions Agreeableness, Conscientiousness, and Emotional Stability; the other, the dimensions Extraversion and Intellect. Two models, one for children and adolescents, the other for adults, were tested by confirmatory factor analysis with generally excellent results. Many personality theorists appear to have considered one or both of these 2 metatraits, provisionally labeled alpha and beta. PMID- 9418279 TI - Role of hope in academic and sport achievement. AB - Hope is the sum of goal thoughts as tapped by pathways and agency. Pathways reflect the perceived capability to produce goal routes; agency reflects the perception that one can initiate action along these pathways. Using trait and state hope scales, studies explored hope in college student athletes. In Study 1, male and female athletes were higher in trait hope than nonathletes; moreover, hope significantly predicted semester grade averages beyond cumulative grade point average and overall self-worth. In Study 2, with female cross-country athletes, trait hope predicted athletic outcomes; further, weekly state hope tended to predict athletic outcomes beyond dispositional hope, training, and self esteem, confidence, and mood. In Study 3, with female track athletes, dispositional hope significantly predicted athletic outcomes beyond variance related to athletic abilities and affectivity; moreover, athletes had higher hope than nonathletes. PMID- 9418280 TI - Compromises produced by the dialectic between self-verification and self enhancement. AB - Three studies of people's reactions to evaluative feedback demonstrated that the dialectic between self-enhancement and self-verification results in compromises between these 2 motives, as hypothesized in cognitive-experiential self-theory. The demonstration was facilitated by 2 procedural improvements: Enhancement and verification were established by calibrating evaluative feedback against self appraisals, and degree of enhancement and of verification were varied along a continuum, rather than categorically. There was also support for the hypotheses that processing in an intuitive-experiential mode favors enhancement and processing in an analytical-rational mode favors verification in the kinds of situations investigated. PMID- 9418281 TI - Effects of variable selection on the factor structure of person descriptors. AB - Previous factor-analytic studies of lexical person descriptors have produced some recurrent patterns of results, but their integration has been hampered by divergences in variable sampling, such as disparate criteria for what is considered a personality descriptor. To isolate effects of variable selection on factor structures, 500 of the most familiar English person descriptors were identified. Fifteen judges provided reliable classifications of these adjectives as disposition, state, social evaluation, or physical-appearance terms. Analyses of adult self-ratings (N = 700) and acquaintance ratings (N = 201) led to a stable Big Five structure when disposition terms, or combined disposition and state terms, were analyzed. Including a wider range of terms led to two additional stable factors: Attractiveness and a factor resembling Big Seven Negative Valence. A stable 3-factor solution was relatively impervious to variable-selection effects. PMID- 9418282 TI - Distinguishing optimism from pessimism in older adults: is it more important to be optimistic or not to be pessimistic? AB - Confirmatory factor analysis revealed that the Life Orientation Test (LOT) consisted of separate Optimism and Pessimism factors among middle-aged and older adults. Although the two factors were significantly negatively correlated among individuals facing a profound life challenge (i.e., caregiving), they were only weakly correlated among noncaregivers. Caregivers also expressed less optimism than noncaregivers and showed a trend toward greater pessimism, suggesting that life stress may affect these dispositions. Pessimism, not optimism, uniquely predicted subsequent psychological and physical health; however, optimism and pessimism were equally predictive for stressed and nonstressed samples. By exploring optimism and pessimism separately, researchers may better determine whether the beneficial effects of optimism result from thinking optimistically, avoiding pessimistic thinking, or a combination of the two. PMID- 9418283 TI - Evaluating stability and change in personality and depression. AB - Critics have argued that personality factors believed to represent a vulnerability to depression are not stable and are therefore state dependent. However, conclusions regarding the stability of personality and the relation between personality and depression have been drawn (a) without differentiating relative stability among individual differences from absolute stability of change scores and (b) without explicitly modeling change in personality as a function of change in depression. The relation between neuroticism and depression was examined in a sample of depressed outpatients (N = 71) receiving a 5-week trial of pharmacotherapy. Measures of neuroticism and extraversion demonstrated both relative stability and absolute change, and changes in neuroticism and extraversion scores were modestly or not at all accounted for by changes in depression scores. Claims that personality scores are not stable and are state dependent must be reconsidered. PMID- 9418284 TI - Competitiveness mediates the link between personality and group performance. AB - The performance of individuals within groups, and of groups as units, is the product of immediate goal structures and personality differences pertinent to those goals among group members. A level-of-analysis approach linked the dimension of agreeableness to situated competitiveness and task performance in group settings. Hypotheses were (a) individual differences in self-rated and other-rated competitiveness are related (inversely) to the Big Five dimension of agreeableness, (b) immediately situated promotive and contrient goal structures influence self-ratings of competitiveness, (c) immediate goal structures differentially activate competitiveness to affect task performance in groups, and (d) agreeableness effects on task performance are partially mediated by competitiveness. Structural equation modeling corroborated hypotheses about the links among agreeableness, competitiveness, and task performance. PMID- 9418285 TI - Working models of attachment and daily social interactions. AB - This study tested whether working models of attachment guide how people construe and respond to social interactions by examining immediate responses to a range of everyday interactions and to specific attachment-relevant interactions. Patterns for immediate reports were compared with those for more memory-based, global reports. Secure, preoccupied, fearful, and dismissing participants provided immediate reports after their social interactions for 1 week and completed retrospective questionnaires. Attachment differences were accentuated in attachment-relevant, high-conflict interactions. Preoccupied participants responded more favorably after conflict than did secure or dismissing-models contribute to perceptions may depend on the fit between attachment goals and the situation and on the extent of memory-based processing. PMID- 9418286 TI - Characterization of cell cultures derived from Fugu, the Japanese pufferfish. AB - The Japanese pufferfish (genus Fugu), which possesses a highly compact genome, is becoming a popular model among those interested in sequencing and mapping the genomes of higher vertebrates. Although genomic libraries have been derived and used to study the molecular biology of Fugu, biological material derived from the living organism is difficult to obtain for laboratories distant from the Asian Pacific. We have established cell cultures from two Fugu species: kusafugu, Fugu niphobles, and torafugu, F. rubripes. Cultures derived from F. niphobles fry and F. rubripes eye have been passaged more than 60 times over the course of one year, representing approximately 180 population doublings. Proliferating cultures were also initiated from F. rubripes brain, liver, fin, spleen, kidney, swimbladder, and muscle. Karyotype analyses indicated that F. rubripes eye derived cells possessed a chromosome number in the diploid range; F. niphobles fry cells were slightly hyperploid. Flow cytometry confirmed that the relative amounts of DNA present in cultured cells from both Fugu species were similar to that measured in blood cells collected from F. rubripes, and approximately one seventh of that measured in diploid human cells. Telomerase activity was easily detectable in lysates prepared from F. niphobles fry cells and F. rubripes eye cells, consistent with the notion that these cultures are capable of indefinite proliferation. PMID- 9418287 TI - Pantropic retroviral vector integration, expression, and germline transmission in medaka (Oryzias latipes). AB - Pantropic retroviral vectors were used to introduce transgenes into Japanese medaka (Oryzias latipes). These vectors contain the long terminal repeat (LTR) sequence of Moloney murine leukemia virus (Mo-MLV) and a reporter gene (neo or lacZ) regulated by the LTR sequence of rous sarcoma virus (RSV). Because these pseudotyped retroviral vectors contain the vesicular stomatitis virus envelope glycoprotein (VSV-G), they have an extremely broad host cell range and can infect many no mammalian species. Newly fertilized medaka eggs (intact or dechorionated) were electroporated at different voltage settings in the presence of 4 x 10(4) cfu of pantropic retroviral vector. The survival rates of the pantropic retroviral vector-treated embryos ranged from 65% to 20% with increasing amplitude of electroporation. Dechorionation did not substantially affect the survival rate of embryos. PCR amplification demonstrated proviral sequences in up to 60% of the 2-month-old fish. The efficiency of gene transfer was enhanced by dechorionation. Furthermore, overnight incubation of dechorionated embryos with pantropic retroviral vectors without electroporation also resulted in proviral integration in 60% of the embryos without compromising survival rate. Southern blot analysis of DNA samples isolated from polymerase chain reaction (PCR) as positive F1 reaction animals confirmed the integration of a single copy of the provirus into the host genome. Three P1 transgenic females transmitted the proviral sequence to 50% of their F1 progeny in a back cross with wild-type males, suggesting that the entire germline of these P1 fish was transformed by the pantropic retroviral vector. Expression of the neomycin phosphotranferase transgene in F1 transgenic individuals was detected by reverse transcription (RT) PCR amplification of the neo mRNA sequence. Furthermore, expression of a beta galactosidase transgene was also observed in 4-day-old F1 transgenic individuals. Thus, pantropic retroviral vectors provide a convenient method to stably introduce and express foreign genes in medaka. PMID- 9418288 TI - Cathepsin, a major protease of the marine sponge Geodia cydonium: purification of the enzyme and molecular cloning of cDNA. AB - Sponges are suspension-feeders that are devoid of body cavities. Phagocytosis is the major route of nutrition in these animals. In an attempt to understand protein digestion, cathepsin was identified in crude extracts from the sponge Geodia cydonium. This enzyme was purified from lysosomes by a two-step procedure- pH precipitation and FPLC separation--to apparent homogeneity; it showed an M(r) of 26,000. Inhibitor as well as substrate studies showed that the sponge cathepsin belongs to the subfamily L of these cysteine proteases. The complete cDNA coding for cathepsin L was isolated and characterized. The deduced aa sequence contains 322 residues, has an M(r) of 36,085, and shows the characteristic signatures known from other cathepsins of the L subfamily: e.g., cleavage site for the proregion, the ERFNIN motif, and the conserved regions forming the catalytic triad of cysteine proteases. Phylogenetic analyses revealed that the sponge sequence groups with the cathepsin L subfamily and branches off first from the other metazoan members. The sponge sequence shows high homology to that isolated from Dictyostelium discoideum and only low similarity to the protozoan cathepsins L from Paramecium tetraurelia and Tetrahymena thermophila. From the data presented it is concluded that cathepsin L is the major digestive protease in sponges. PMID- 9418289 TI - Characterization and expression of c-type lysozyme cDNA from Japanese flounder (Paralichthys olivaceus). AB - Lysozyme is a widely distributed enzyme located in the serum, skin mucus, and other organs of fish, which is responsible for catalyzing the hydrolysis of the cell walls of most bacteria. A c-type of lysozyme cDNA was cloned from a kidney cDNA library of the Japanese flounder (Paralichthys olivaceus). The cDNAs consisted of 612 bp, which coded for 143 amino acid residues. The deduced amino acid sequence of Japanese flounder c-type lysozyme possessed 72.9%, 57.4%, and 65.4% identities with rainbow trout, chicken, and human c-type lysozymes, respectively. Comparison of the c-type lysozymes showed that the catalytic residues, the residues binding to sugar chains, and cysteine residues were completely conserved. Northern blot analysis indicated that the c-type lysozyme gene is apparently transcribed in the head kidney, posterior kidney, spleen, brain, and ovary of healthy flounder. When flounder were experimentally infected with Edwardsiella tarda, quantities of the c-type lysozyme mRNA increased in the head kidney, spleen, and ovary of the flounder. PMID- 9418290 TI - Expressed sequence tags of medaka (Oryzias latipes) liver mRNA. AB - A medaka liver cDNA library was constructed in lambda ZAPII. The number of clones in this library is approximately 5 x 10(6). Three hundred sixty-one clones were randomly selected and, of these, 33 clones with over 1 kb were sequenced. These sequences were compared with GenBank and the dbEST (Re.96.0). Twenty-five clones of these 33 clones encoded 18 different genes and 2 different expressed sequence tags (ESTs). Sequences of 10 of the 18 clones had not previously been reported in fish. The codon usage of medaka genes is similar to that of Xenopus, mouse, and human genes, but not similar to that of yeast and Escherichia coli genes. PMID- 9418291 TI - Cloning and characterization of transferrin cDNA and rapid detection of transferrin gene polymorphism in rainbow trout (Oncorhynchus mykiss). AB - A cDNA clone of rainbow trout (Oncorhynchus mykiss) transferrin was obtained from a liver cDNA library. The 2537-bp cDNA sequence contained an open reading frame encoding 691 amino acids and the 5' and 3' noncoding regions. The amino acid sequences at the iron-binding sites and the two N-linked glycosylation sites, and the cysteine residues were consistent with known, conserved vertebrate transferrin cDNA sequences. Single N-linked glycosylation sites existed on the N- and C-lobe. The deduced amino acid sequence of the rainbow trout transferrin cDNA had 92.9% identities with transferrin of coho salmon (Oncorhynchus kisutch); 85%, Atlantic salmon (Salmo salar); 67.3%, medaka (Oryzias latipes); 61.3% Atlantic cod (Gadus morhua); and 59.7%, Japanese flounder (Paralichthys olivaceus). The long and accurate polymerase chain reaction (LA-PCR) was used to amplify approximately 6.5 kb of the transferrin gene from rainbow trout genomic DNA. Restriction fragment length polymorphisms (RFLPs) of the LA-PCR products revealed three digestion patterns in 22 samples. PMID- 9418292 TI - A functional study of the salmon GnRH promoter. AB - The Pa and Pb promoters of the Atlantic salmon (Salmo salar) GnRH gene were fused together or individually to the Escherichia coli lacZ gene, and their transcriptional activities were measured in transient expression assays in zebrafish (Danio rerio). In 48-hour embryos, both promoters were preferentially expressed in the brain, whereas a cytomegalovirus (CMV) promoter-lacZ fusion gene displayed high levels of activity in nonbrain tissues. Pa and Pb exhibited different cell specificity in the forebrain. Pb was active in large neuron-like cells exclusive in the olfactory placode region, whereas Pa appeared active in nonneuron-like cells in the forebrain. In Atlantic salmon forebrain tissue, both Pa and Pb exhibited endogenous activity, as assessed by reverse transcriptase polymerase chain reaction (RT-PCR) analysis. However, only the Pb transcript contained the prepro-GnRH exon II-IV sequences, suggesting that Pa activity may not be related to GnRH production in this species. PMID- 9418293 TI - Characterization of transgenic tilapia lines with different ectopic expression of tilapia growth hormone. AB - The transfer of growth hormone (GH) genes has opened new possibilities for the manipulation of growth in economically important fish species. However, the ectopic GH levels to optimize growth acceleration in fish, and specially in tilapia, are not known and must be determined experimentally. The tilapia GH (tiGH) cDNA was used to construct chimeric genes expressing different levels of tiGH in vitro and in vivo. These constructs were used to generate four lines of transgenic tilapia by microinjection into one-cell embryos. Different patterns and levels of ectopic expression of tiGH and IGF were detected in organs of transgenic tilapia by RNA or protein analysis. The two lines with lower ectopic tiGH mRNA levels were the only ones showing growth acceleration, suggesting that the expression of ectopic tiGH promoted growth only at low expression levels. The effect of higher ectopic tiGH levels resembled the physiologic situation of low condition factor and permitted us to postulate a model for growth acceleration in transgenic tilapia expressing ectopic tiGH. PMID- 9418294 TI - A survey of expressed genes in Japanese flounder (Paralichthys olivaceus) liver and spleen. AB - Expressed sequence tags (ESTs) were obtained from Japanese flounder (Paralichthys olivaceus). We present the results of single-pass sequencing of 493 ESTs from 350 clones from liver cDNA and 57 ESTs from 41 clones from spleen cDNA. Sequences of the cDNA clones were compared with sequences in the GenBank database. Two hundred and two clones (51.7%) appeared to be completely unknown and are likely to represent newly described genes, whereas 189 clones (48.3%) were identified based on matches to sequences in the databases. Three of the unidentified sequences were isolated from both the liver and spleen cDNA libraries. However, there were no identical sequences between liver and spleen clones. PMID- 9418295 TI - An efficient DNA extraction method for small metazoans. AB - The isolation of total nucleic acids from small metazoan taxa is difficult and often leads to an unacceptably large percentage of unsuccessful polymerase chain reaction (PCR) amplifications. Our work with the evolutionary genetics of harpacticoid copepods was an incentive to refine techniques such that consistent amplifications from minute marine organisms were feasible. We describe these modifications and demonstrate their utility for the amplification of multiple loci from single harpacticoid copepods. PMID- 9418296 TI - Isopentenyl diphosphate isomerase: a core enzyme in isoprenoid biosynthesis. A review of its biochemistry and function. PMID- 9418297 TI - Indolizidine and quinolizidine alkaloids. PMID- 9418298 TI - The 1997 Nobel Prizes in Science. PMID- 9418299 TI - Off with its head! PMID- 9418300 TI - Bacterial gene swapping in nature. PMID- 9418301 TI - The placebo effect. PMID- 9418302 TI - Novel vaccine technologies. PMID- 9418303 TI - G-protein-coupled receptors in Saccharomyces cerevisiae: high-throughput screening assays for drug discovery. AB - G-protein-coupled receptors are an important class of therapeutic drug targets by virtue of their roles in the regulation of diverse cellular functions. Recent advances in the expression of heterologous G-protein-coupled receptors in the yeast Saccharomyces cerevisiae have led to the development of sensitive and selective assays of their ligand-induced activation. Implementation of this new technology in the high-throughput screening of compound libraries has enabled the discovery of novel ligands for the G-protein-coupled somatostatin receptor. This article describes the broad applicability of the technology and its use in drug discovery. PMID- 9418304 TI - Bacterial biosensors for monitoring toxic metals. AB - Biosensors utilize biological components to provide selectivity for monitoring compounds of environmental, clinical and industrial importance. A number of biosensors based on bacteria have recently been developed for monitoring toxic metals in the environment. The advantages and disadvantages of these types of biosensors are discussed. PMID- 9418305 TI - Ultrasonic separations in analytical biotechnology. AB - Cells in megahertz-frequency noncavitating ultrasonic standing waves concentrate at submillimetre distances and are, as large clumps, easily removed from suspension in flow or batch systems. An ultrasonic filter for perfusion hybridoma culture is now available. Results from small-volume prototype analytical-scale systems can inform the design of effective filter or batch-clarification systems for a wide range of cell sizes, concentrations and sample volumes. Large increases in the rates of aqueous biphasic separations and of the rates and sensitivities of analytical immunocoated particle-agglutination assays occur in standing waves. Ultrasonic manipulation is briefly compared with immunomagnetic and dielectrophoretic separations. PMID- 9418306 TI - The genetic reconstruction of BCG as a new immunotherapeutic tool. AB - The bacillus of Calmette and Guerin (BCG), long appreciated for its role as a live vaccine for the prevention of tuberculosis, is undergoing a rebirth as a recombinant delivery vehicle for foreign antigens and bioactive proteins. Recombinant BCG causes long-lived specific humoral and cellular immunity and may ultimately prove to be a powerful and cost-effective new weapon against both infectious pathogens and certain cancers. PMID- 9418307 TI - Directed evolution of enzyme catalysts. AB - Directed enzyme evolution has emerged in the past few years as a powerful alternative to rational approaches for engineering biocatalysts. Prerequisites for successful directed evolution are functional expression in a suitable microbial host, a rapid screen for the desired feature(s) and a well-thought-out working strategy for navigating protein landscapes. The rapidly growing body of literature on enzyme evolution in vitro includes techniques for creating and searching combinatorial enzyme libraries, as well as several successful examples of different evolutionary strategies being used. PMID- 9418308 TI - [Limited axillary thoracotomy for recurrent spontaneous pneumothorax]. AB - Recurrent spontaneous pneumothorax often requires surgical intervention. Recently, less invasive thoracic surgical techniques, such as video-assisted thoracoscopy (VAT) and limited axillary thoracotomy (LAT), have been developed and used for different thoracic procedures. We describe our results with limited axillary thoracotomy, as compared with those of video-assisted thoracoscopy as reported in the literature. From October 1994 to May 1996, 14 patients with recurrent spontaneous pneumothorax, aged 16-33 years, underwent limited axillary thoracotomy, resection of blebs and apical pleurectomy, using multifire GIA 80 staplers (Auto Suture Inc.). There were no complications or recurrences during 5 17 months of follow-up. Mean operative time was 52.2 minutes and mean hospital stay 2.3 days postoperatively. Full activity was regained within 12.1 days. In comparison with over 75 cases of VAT from the literature, LAT is safe and offers the potential benefits of decreased operative time, hospital stay and cost. PMID- 9418309 TI - [Carnitine deficiency in inborn errors of metabolism]. AB - Several conditions, considered as inborn errors of metabolism, involve severe deficiencies in carnitine in both plasma and muscle. In the absence of evidence suggesting primary carnitine deficiency due to a biosynthetic enzymatic defect in the liver, the various diseases with carnitine deficiency are related to genetic defects in organic acid metabolism leading to blocked mitochondrial beta oxidation. We describe a 4.5-year-old boy and 2 female infants with glutaric aciduria type I, isovaleric acidemia, and long-chain acid dehydrogenase deficiency, in whom severe carnitine deficiency was apparent. In all 3, long-term carnitine treatment proved to be vital and eliminated most of the symptoms. PMID- 9418310 TI - [Penetrating ocular injuries: an epidemiologic and retrospective study]. AB - Penetrating injuries of the eye are an important cause of unilateral visual loss. We studied a series of 82 cases of penetrating injuries treated here from 1987 through 1993. The injuries were caused by sharp objects in 66% and blunt trauma in 6%. The prognosis after a penetrating injury is greatly influenced by the nature of the injury and the extent of the initial drainage. Among factors associated with an unfavorable visual outcome were diminished preoperative visual acuity and scleral wounds with dense vitreous hemorrhage. PMID- 9418311 TI - [Identification of sentinel and axillary node involvement in breast cancer]. AB - Axillary node dissection for breast cancer is important for staging and prognosis. "Sentinel nodes" are the first nodes into which primary cancer drains. Identification, removal and pathological examination of those nodes indicates whether completion of axillary lymphadenectomy is required. The sentinel nodes are identified using a vital dye injected at the primary tumor site. With this technique we were able to identify sentinel nodes in 46 of 48 (95%) women examined. An average of 2.7 +/- 1.2 nodes were identified as sentinel nodes. In 81% of cases there was a correlation between involvement of sentinel nodes and of other axillary nodes as well. In 10% of patients sentinel nodes were involved with tumor while other axillary nodes were negative. The major problem in routine application of this is relationship in surgical decisions is reliable real time pathological identification of lymph node involvement by tumor. PMID- 9418312 TI - [Analgesia in breast surgery with interpleural bupivacaine]. AB - A control group of 15 patients undergoing breast surgery was given general anesthesia. In 15 other patients an interpleural block with 0.4 ml/kg bupivacaine, 0.5%, was performed 20 minutes before induction of general anesthesia for pre-emptive analgesia. This was extended further by continuous administration of bupivacaine 0.25%, 0.125 ml/kg/hr by automatic infusion pump, with supplements of opiates for postoperative pain management. The combined technique was associated with significantly reduced perioperative opiate requirement with better emergence from anesthesia, fewer side effects, a prolonged pain-free period, and overall better quality of postoperative recovery. PMID- 9418313 TI - [Choanal atresia: 13 years of experience]. AB - Choanal atresia is uncommon and consists of congenital blockage between the nasal cavity and the nasopharynx. The anomaly presents either immediately after birth as respiratory distress, or as a coincidental finding at an older age. Treatment is usually surgical. The approaches are transnasal, transseptal, transpalatal and transantral. Different types of stents are used and for various periods after each type of correction. Between 1983-1996, 20 patients with choanal atresia were operated on, in 12 of whom it was bilateral. The youngest was 3 days old and the oldest 22 years (average 6 years). The 20 patients underwent a total of 29 operations of which all were transnasal except for 2 corrected through a transseptal approach; 3 had their primary operation elsewhere. In all cases the atresia was bony or combined bony and membranous, except for 2 in whom there was combined atresia on 1 side and membranous on the other. The success rate was 75% in those first operated on here, in whom stents were employed. In our last 5 cases we used the endonasal approach and a rigid endoscope, a safe technique that has the advantage of direct unobscured vision. PMID- 9418314 TI - [Fear of personal death among hospital physicians]. AB - Many studies have tried to explain why professionals experience difficulty when dealing with, and in treating efficiently situations connected with death. We studied levels of personal fear among physicians in general hospitals and addressed 2 questions: Does exposure to death on professional and personal levels, affect the level of the fear of personal death which physicians experience? Is there a relationship between personality variables, represented by the repression-sensitization dimension, and level of fear of personal death? A sample of 233 physicians from 22 general hospitals who specialized in oncology, internal medicine, surgery, psychiatry or pediatrics was studied. Each answered 4 questionnaires with regard to demographic information, fear of personal death, level of repression-sensitization and exposure to the death of relatives and significant others. There were no differences in level of fear of personal death of physicians according to specialization, but those who had been exposed to death on the personal level, feared less their own death. With respect to the personality variable, tendency to sensitization, it was found that those who were sensitized exhibited a higher level of the fear of their own death compared to those who were repressive. Of the various demographic variables examined (sex, level of religious observance, age, number of children, health, professional experience) it was found that those: with many years of professional experience, who were relatively older, who were nonobservant religiously and who were in good health, had lower levels of personal fear of death; gender was not a factor. PMID- 9418315 TI - [Kleptomania: phenomenological, clinical and legal aspects]. AB - Kleptomania is currently classified in psychiatric nomenclature as one of the impulse control disorders (DSM-IV, 1994). It is characterized by repeated failure to resist impulses to steal objects, not for personal use or monetary gain. The objects are therefore discarded, given away, or hoarded (ICD-10, 1992). This disorder is known since the early 18th century from the phenomenological and clinical viewpoints, yet is still debated with regard to therapeutic strategies and criminal liability. Although there are usually complications associated with the legal consequences of being caught and arrested, subjects continue to violate the law despite repeated arrests and convictions. In a 28-year old man suffering from kleptomania, years of psychodynamic psychotherapy were ineffective. Only when he was treated as suffering from an impulse control disorder or a variant of obsessive-compulsive disorder, was there significant improvement. The positive response to buspirone (5-HT1A) augmentation of fluvoxamine (SSRI) suggested that disturbed central serotonergic neurotransmission might play an important role in the pathogenesis of kleptomania. This concept is strengthened by the comorbidity of the syndrome with depression and by its compulsive traits. We stress that although kleptomaniacs cannot differentiate between right and wrong, testing shows that their sense of reality is intact, but they act under the influence of drives they cannot resist. PMID- 9418316 TI - [Infection by Vibrio vulnificus after a prick from from the spine of a Tilapia]. AB - Vibrio vulnificus is a Gram-negative bacterium living in warm salty water that produces a spectrum of human disease which may progress to devastating, sometimes fatal infections in susceptible individuals. Such infections have rarely been reported in Israel. However, over the past few months we have been seeing a sharp increase in V. vulnificus infections with a common history of injury to extremities by the sharp spines of Tilapia zillii, ("amnon" or St. Peter's fish). Clinical suspicion and prompt intervention prevent the untoward consequences of misdiagnosis or delay. PMID- 9418317 TI - [Fatal multiple organ system dysfunction associated with germanium metal used in complementary therapy]. AB - The element germanium is widely distributed in nature. It is used in industry as a semiconductor and there have been a few attempts to use it in medicine. In the past few years 20 patients have been described in the literature as suffering from germanium overdosage. Like laboratory animals affected by the element, they suffer from renal failure and injury to other organs. We describe a 52-year old man given germanium to prevent recurrence of a brain tumor. He developed multiple organ dysfunction syndrome and died of intractable hyperdynamic shock. We call for caution regarding morbidity resulting from treatments believed safe. PMID- 9418319 TI - [Withdrawal of anticonvulsive therapy: when, how, and in which patient?]. PMID- 9418318 TI - [Traumatic hyphema]. AB - Traumatic hyphema usually occurs in young men at the rate of 17-20/1000,000. Major complications include secondary hemorrhage, glaucoma, corneal staining and disturbances in visual acuity. Final visual acuity is predominantly the outcome of all the ocular injuries occurring during the trauma, mainly to the posterior segment of the eye. We describe all cases of traumatic hyphema treated in our department over a period of 3.5 years. Antifibrinolytic treatment is recommended in the literature in traumatic hyphema to prevent secondary hemorrhage. Our findings differ from those in the literature in that they show a lower prevalence of more severe hemorrhages and of secondary hemorrhage. In light of these differences, and with regard to possible side effects of such treatment, we suggest that antifibrinolytic treatment not be used in our population. We recommend that treatment for traumatic hyphema should include restricted activity, local corti-costeroidal preparations, frequent follow-up visits and vigorous diagnostic work-up in order to find any additional eye damage. We strongly recommend the use of preventive measures (eye-shields) in high risk activities such as sports, house-hold work and military training. PMID- 9418320 TI - [BRCA1 and BRCA2--breast cancer susceptibility genes]. PMID- 9418321 TI - [Anti-endothelial cell antibodies--clinical significance]. PMID- 9418322 TI - [How much should blood pressure be reduced? Is there a J-shaped relationship between lowered blood pressure and cardiovascular morbidity?]. PMID- 9418323 TI - [Nutritional support for burn casualties]. PMID- 9418324 TI - [Coronary artery aneurysm]. PMID- 9418325 TI - [Surgical treatment of cryptorchidism]. PMID- 9418326 TI - [Syncope induced by cardiovascular drugs]. PMID- 9418328 TI - [The road into the open--Dr. Arthur Schnitzler's story]. PMID- 9418327 TI - [Multisystem injury in the elderly]. PMID- 9418329 TI - [Sources of some medical terms]. PMID- 9418330 TI - [Brain imaging and its clinical application in psychiatry]. AB - The common structural and functional brain imaging techniques are described from a practical, clinical point of view. The clinical indications for brain imaging in psychiatry are reviewed in relation to the specific limitations and advantages of each technique. The clinical applications of computerized tomography (CT), magnetic resonance imaging (MRI) and single photon emission computerized tomography (SPECT) are discussed in relation to the differential diagnosis between organic and functional psychiatric disorders. In a 55-year-old man with late onset of behavioral changes but without neurological signs the application of structural brain imaging (CT and MRI) in case management was demonstrated. The imaging findings involved the differential diagnosis between depression and focal brain lesions. In a 38-year-old man with personality changes and depression following a traumatic brain injury, time interval repeated functional brain imaging (SPECT) was used. Brain imaging reflected improvement in clinical status following treatment and was able to differentiate between reversible and permanent traumatic brain injuries. The superior yield of time interval repeated functional imaging in diagnosis and management of postconcussion syndrome is discussed. PMID- 9418331 TI - [Orthopedic ward policy in introduction of new types of total hip implants]. AB - The use of different types of total hip implants in medical centers in Israel was surveyed. Questionnaires were sent to all orthopedic ward directors in Israel requesting information on the number of total hip arthroplasties performed between the years 1984-1993, the types of implants used, and whether attending physicians or residents perform the operations. 22 of 24 orthopedic wards responded but 1 ward was excluded because only the results for 1993 were reported. 5 wards reported more and 16 fewer than 50 operations a year. 15 different types of implants were in use in Israel in that period, and in 5 wards 5 or more types of implants were used. Only 1 of the wards performed more than 50 operations a year. We conclude that the indiscriminate use of multiple technologies in wards performing few operations can lead to the long "learning curves" previously associated with poor results. Orthopedic surgeons should resist the impulse to introduce new implants, thus improving results and lowering expenditure. The need for regulating the introduction of new implants is emphasized. PMID- 9418332 TI - [Consanguinity among Arabs in Israel]. AB - In a previous nationwide study, we examined the rate of consanguineous matings in the Israeli Arab community. The average inbreeding coefficient was 0.0192, much higher than that reported for the general population of Israel, 0.0038 in 1956-7. The inbreeding coefficients of 69 Arab villages, towns and cities (excluding the Bedouin in the South) were determined. Knowledge of the inbreeding coefficients of the various local populations is of value for geneticists, pediatricians and gynecologists and for planning suitable health programs. PMID- 9418333 TI - [Physician-patient relations--participation of patients with ureteral calculi in clinical decision making and level of anxiety]. AB - In a study examining the relationship between patient participation in clinical decision making and levels of anxiety, patients were offered a choice of treatment for ureteral calculus. 42 received information about 2 treatment options, ultrasound fragmentation of the stone through a ureteroscope and extracorporeal shock wave lithotripsy (ESWL), and were asked to choose the method that they preferred. 54 received treatment decided on by the physician without their participation in the decision making process. Anxiety was measured before meeting with the physician, immediately after the meeting and on hospitalization for treatment. The contribution of the patient's perception of participation in the decision-making process and level of education was also examined. There was a decrease in level of anxiety after meeting with the physician only among those who did not actually participate in the decision-making process (p < 0.05). There was no change in the level of anxiety among those offered choice of treatment. However, a decrease in anxiety was evident among patients who perceived that they had received information about their illness and its treatment (p < 0.01). This was not the case for patients who perceived themselves as participants in decision making unless they had a relatively high-level of education (p = 0.05). PMID- 9418334 TI - [Stromal uterine sarcoma arising from intestinal endometriosis after abdominal hysterectomy and salpingo-oophorectomy]. AB - The incidence of ectopic endometriosis is 4% to 18%. The intestinal type is quite common with the rectosigmoid the most likely part of the bowel to be involved due to its pelvic location. A 43-year-old woman, whose symptoms, X-ray and endoscopic findings suggested malignancy of the rectosigmoid, is presented. Primary malignancy of the bowel was excluded by endoscopic biopsies. Histological examination at operation showed stromal sarcoma of the uterus with foci of endometriosis. There is no report in the English literature of transformation of intestinal endometriosis into malignancy, such as stromal sarcoma of the uterus, which imitates a primary malignancy with obstruction of the rectosigmoid. PMID- 9418335 TI - [Blunt trauma causing emboli from friable atherosclerotic plaques]. AB - We present a 55-year-old woman who developed a shower of emboli following a car accident. Such events may have medicolegal implications as well as preventive considerations. PMID- 9418336 TI - [Combined treatment for adhesive capsulitis of the shoulder]. AB - Adhesive capsulitis is a problem for the orthopedic surgeon due to the difficulty of treatment. Although it is self-limited, few patients will wait for spontaneous resolutions while suffering pain and progressive loss of motion. Our aim was to modify the course of the disease and to shorten recovery time by combining intensive physiotherapy with intra-articular infiltration and gentle manipulation. 49 patients with 50 frozen shoulders were enrolled in the study. All patients were treated initially with physiotherapy for 4-8 weeks. If no improvement was noted the affected shoulder was infiltrated and gently manipulated. 27 of 49 patients (55%) improved dramatically with the initial physiotherapy regimen. They achieved full or nearly full range of motion, with significant relief of pain. 22 patients were infiltrated and manipulated. Elevation improved significantly from an average of 110.95 to 165.71 degrees (p < 0.001), external rotation from an average of 9.52 to 43.57 degrees (p < 0.001) and internal rotation also improved significantly (p < 0.001). Self assisted physiotherapy is the corner stone of treatment in adhesive capsulitis. When pain and limitation of passive range of motion persist, infiltration and gentle manipulation dramatically shortens the debilitating process. PMID- 9418338 TI - [Biofeedback treatment of Raynaud's disease]. AB - Raynaud's disease is characterized by intermittent peripheral vasoconstriction leading to pallor, cyanosis and reactive vasodilation of the arterioles of fingers and toes. These phenomena are accompanied by sensations of cold or warmth, pain and difficulty in manipulating the palms. Ulcerations of the fingertips can occur in severe cases. Since conservative medical treatment, consisting of preventive measures and changing various habits, results in alleviation in only half the patients, sympathectomy is often required. Psychological intervention, including biofeedback, also has a significant role. Biofeedback involving relaxation techniques, guided imagination, and in parallel, computer-assisted monitoring of sympathetic arousal, might lead to symptom reduction as a unique treatment or in conjunction with other treatment modalities. PMID- 9418337 TI - [Macroenzymes: an interesting laboratory finding, without clinical relevance]. AB - Macroenzymes are complexes of serum enzymes with proteins which have a higher molecular weight and longer plasma half-life than the normal enzyme. The presence of macroenzymes is suggested by finding increased serum enzyme activity, not associated with symptoms. Thus, macroenzymes can cause diagnostic errors and the performance of unnecessary tests or invasive procedures. We describe 2 patients with highly elevated serum levels of lactate dehydrogenase (LDH) and creatine kinase (CK) due to formation of complexes with immunoglobulin G. 1 patient had LDH of 4500 u/L but was otherwise normal and in the second CK was elevated with no evidence of ischemic heart disease. Awareness of the phenomenon of macroenzymes may save the patient long and sometimes invasive investigation. PMID- 9418340 TI - [Total hip replacement: do we need to use the whole variety of implants?]. PMID- 9418339 TI - [Isolated susceptibility of Streptococcus pyogenes to 3 macrolides]. AB - Erythromycin is considered the drug of choice in the treatment of streptococcal pharyngitis in patients allergic to penicillin. However, in recent years several publications, especially in Finland and Italy, showed high resistance rates of S. pyogenes isolates to erythromycin and other new macrolides. To evaluate the situation in Israel, we checked the MIC of isolates from patients with tonsillitis during 1996. E-test results showed an MIC-50 of 0.23, 0.13 and 0.47 mcg/ml for erythromycin, clarithromycin and roxithromycin, respectively and a MIC 90 of 0.37, 0.23 and 0.78 mcg/ml. Only 2 isolates (2.1%) were partially or completely resistant to all 3 antibiotics. We conclude that empiric therapy with macrolides in Israel is still a viable option and can be recommended in S. pyogenes tonsillitis for patients allergic to penicillin. PMID- 9418341 TI - [Aortic atheromas as a risk factor for stroke and embolism]. PMID- 9418342 TI - [Ancient mutations in the Sons of Shem cause familial Mediterranean fever]. PMID- 9418343 TI - [Benign and premalignant breast lesions--clinico-pathological correlation]. PMID- 9418344 TI - [Water birth]. PMID- 9418346 TI - [Impact of genetic technology on doctor-patient-society relationships]. PMID- 9418345 TI - [Ozone and its toxic effects]. PMID- 9418347 TI - [Stroke--advances in diagnosis and treatment]. PMID- 9418348 TI - [Amniotic fluid embolism--pathogenesis and management]. PMID- 9418349 TI - [Blunt trauma during pregnancy]. PMID- 9418350 TI - [VIP in the emergency department--a "one man disaster", proposal of treatment protocol]. PMID- 9418351 TI - [Quality of life in younger adults (17-49) after first stroke--a two year follow up]. AB - To study the effect of stroke on the quality of life in younger adults, 199 patients 17-49 years of age who had sustained a first stroke between 1.11.92 and 31.10.93 were followed up. They were interviewed by telephone at 3, 6, 12 and 24 months after the event. 2 died during the first year of follow-up, and 8 had recurrent strokes. After 2 years, 8 additional patients had died and 4 had sustained recurrent events. Gradual improvement was reported within all age groups and in all areas. During the 3-6 months period, a mean of 4% improvement occurred in functional capability, 15% in social and recreational activity and 8% in return-to-work. The 6-12 month period showed an increase of 3% in improvement in mean functional capability, 10% in social and recreational activity and 2% in return-to-work. 1 year after the stroke 27% remained with moderate to severe disability, but over 86% were functionally independent in their daily living activities. There were no significant changes during the second year of follow-up in these statistics. 67% of those employed prior to their stroke returned to work and approximately 70% reported a return to prestroke social and recreational activity. These results demonstrate that the relatively high recovery rate and functional improvement during a year of follow-up were not accompanied by similar rates of improvement in employment and in social integration. They indicate the need for increased emphasis on long-term psychosocial rehabilitation services within the community. PMID- 9418352 TI - [Epidemiological characteristics of outbreaks of diarrhea and food poisoning in the Israel Defense Forces in the years 1978-1995]. AB - Acute infectious diseases of the gastrointestinal tract and food poisoning are problems of great importance in the Israel Defense Forces (IDF). They involve individual and epidemic morbidity, with impairment of health of individual soldiers and in the activities of units. Outbreaks of gastrointestinal infectious diseases must be reported to the IDF army health branch, which conducts epidemiological investigation. This study is based on data from yearly epidemiological reports for 1978-1989, and from a computerized database for the years 1990-1995. The incidence of outbreaks is characterized by an unstable trend. It was highest at the end of the 80's (68.3 per 100,000 soldiers on active duty) and lowest for the last 2 years (1994-1995, 36.3 per 100,000). The incidence of soldiers involved in food-borne outbreaks has been more stable, constantly declining during the course of the years. There was marked seasonality with a peak in the summer months. Sporadic morbidity was constant in 1990-1995, with a yearly attack rate of 60% in soldiers on active duty. Shigella strains were the leading cause of outbreaks until 1993, while in 1994-1995 their proportion decreased, with an increase in the proportion of Salmonella strains. As to Staphylococcus aureus, its role in causing food poisoning has been characterized by marked changes. Shigella sonnei replaced Shigella flexneri as the leading strain. 73.3% of outbreaks were small, with fewer than 40 soldiers involved, while 5.4% of outbreaks affected more than 100 soldiers. Outbreaks in which a bacterial agent was identified or which occurred in new-recruit bases were larger than those in which a bacterial agent was not identified, or which occurred in active field unit bases. In conclusion, the rates of infectious disease of the gastrointestinal tract are still high, although there has been a marked decrease since 1994. The incidence of outbreaks has also decreased, as well as the role of Shigella as a leading causative agent. PMID- 9418353 TI - [Has the health profile and pregnant Ethiopian immigrants changed in the Ashkelon region in the past years?]. AB - There are about 6500 births yearly in the Ashkelon District. 6% of the mothers are Ethiopians, most of whom immigrated to Israel since the early 90's. Our data are from 3 sources: birth certificates, infant death certificates, and the national population register. Birth rates in single mothers and rates of low birth-weight births have declined over the years. Infant mortality and still birth rates have also shown remarkable and consistent declines between 1990-1995. We conclude that improvement in life conditions of Ethiopian immigrants and better use of health services have had a great impact on birth outcomes and pregnancy patterns. The gap between Ethiopian immigrants and other Jewish communities in Israel is closing fast. PMID- 9418354 TI - [Advances in ophthalmological photodynamic therapy]. AB - Photodynamic therapy is a new experimental therapeutic technique which is attracting increasing attention. Its biopharmacological basis of action is the specific interaction of a photosensitizing compound with the cellular elements of pathological lesions. The photosensitizer is thought to enter specifically into the pathologic cells, where it accumulates. The lesion is then irradiated with a sensitizing laser-beam of specific wave-length to activate the photosensitizer, which then becomes a generator of free oxygen radicals. These radicals destroy the sensitizer-harboring pathological cells. The advantage of specifically destroying pathological lesions without affecting surrounding normal tissue is obvious. Recently, many experimental studies have been conducted to test the usefulness of photodynamic therapy for ocular disorders, mainly advanced age related macular degeneration and uveal melanoma. Results so far are encouraging. PMID- 9418355 TI - [Occupational accidents and eye injuries treated at the Rambam Medical Center]. AB - To determine the prevalence of occupation-related eye injuries, we analyzed the records of 24,632 patients treated at our emergency surgical department over a 3 year period. Occupational accidents accounted for 17.6% of such cases. A third of them (1374 patients) were referred to the ophthalmic emergency room for further examination. In 89% (1223) of these, at least 1 pathological ocular finding was detected, and 8.3% (114) of them were hospitalized. Penetrating eye injuries were found in 5.2% (72). The commonest eye injury was corneal foreign body found in 42.8%. PMID- 9418356 TI - [Gram-negative enteric bacteremia in children in the Negev (1989-1994)]. AB - During 1989-1994, there were 322 episodes of Gram-negative enteric bacteremia in 308 children. The incidence increased from 31/100,000 in children younger than 15 years of age during 1989-1991, to 50/100,000 during 1992-1994. The most common pathogens were Klebsiella, E. Coli, Salmonella and Enterobacter. 39% of episodes were nosocomial and a significant increase was recorded for each species during the last 3 years of the study. Klebsiella represented the most common pathogen causing nosocomial bacteremia, while E. coli and Salmonella were the main pathogens causing community-acquired bacteremia. In this study in southern Israel, the incidence of Gram-negative enteric bacteremia was significantly higher in Bedouin children, with the exception of bacteremia due to Salmonella, which occurred mainly in Jewish children. PMID- 9418357 TI - [Q fever endocarditis and bicuspid aortic valve]. AB - Q fever is caused by the rickettsia Coxiella burnetti, an obligate intracellular bacterium acquired by inhalation of infected dust from subclinically infected animals. Q fever may be acute or chronic; the chronic form mostly presents as endocarditis. Immunocompromised states and underlying heart disease are the most important risk factors. Usually the symptoms of Q fever endocarditis are nonspecific and diagnosis is often established very late. New criteria for diagnosis include a single blood culture positive for Coxiella burnetti, positive Q fever serology and characteristic echocardiographic studies. We describe a 49 year-old man with bicuspid aortic valve admitted with fever, weight loss and a new heart murmur. The diagnosis of Q fever endocarditis was established by positive Q fever serology, and an echocardiogram showing vegetations and valvular dysfunction. This case suggests that Q fever endocarditis should be considered in patients with "sterile" endocarditis. PMID- 9418358 TI - [Gradenigo syndrome and cavernous sinus thrombosis in fusobacterial acute otitis media]. AB - In this era of antimicrobial medication, intracranial complications following otitis media are rare. We present a 5-year-old boy who suffered from petrositis (Gradenigo syndrome) and cavernous sinus thrombosis as combined complications of acute otitis media caused by fusobacteria. The diagnosis was made using imaging methods suited to the various structures of the skull. Cure was achieved by prolonged conservative treatment with antibiotics, with gallium scan for evaluation of the bone inflammation. PMID- 9418359 TI - [The changing world]. PMID- 9418360 TI - [The genetic passport--too much, too little]. PMID- 9418362 TI - [Erectile dysfunction--practical guidelines for evaluation]. PMID- 9418361 TI - [Cardiac rehabilitation]. PMID- 9418363 TI - [Erectile dysfunction--practical guidelines for treatment]. PMID- 9418364 TI - [P21ras--a central cross-road on intracellular signalling]. PMID- 9418365 TI - [Hormonal changes in term and preterm delivery]. PMID- 9418366 TI - [Acute glaucoma as a complication of nebulized bronchodilators]. PMID- 9418367 TI - [Chronic management of cor pulmonale]. PMID- 9418368 TI - [Bacterial (pyogenic) sacroiliitis]. PMID- 9418369 TI - [Position paper on physician-patient relationships]. PMID- 9418370 TI - [Nonoperative management of splenic injuries in children: guidelines for aftercare]. PMID- 9418371 TI - [Intraperitoneal adhesions]. PMID- 9418372 TI - [Colonic obstruction and bilateral hydroureter related to periappendicular abscess]. PMID- 9418373 TI - Physiological perspectives of copper. AB - Living organisms are composed of only 30 elements, nine of which are required in traces (1:20,000), hence called "Trace elements". The family of trace elements comprises of selenium, chromium, manganese, iron, cobalt, copper, zinc, molybdenum and iodine. Each element has specific associations referred as metalloenzymes. PMID- 9418374 TI - Photoperiod and melatonin-induced changes in male reproduction in Indian desert gerbil, Meriones hurrianae (Jerdon). AB - Exposure to continuous darkness and chronic treatment with melatonin, for six weeks, stimulated reproduction in the male Indian desert gerbil as evidenced by morphometric data. Exposure to continuous light, for same duration, on the other hand inhibited reproduction. The results are opposite to those reported from similar studies on temperate zone species. Surface areas of abdominal scent glands increased following both, exposures to continuous darkness and the treatment with melatonin. Exposure to continuous light decreased the scent gland surface area. Assessment of scent gland activity could be useful in evaluation of reproductive function as they are dependent on sex steroids. Melatonin that mediates photoperiodic influence on reproduction is not always inhibitory to gonads. PMID- 9418375 TI - Comparative in vitro and in vivo evaluation of himachalol in murine invasive aspergillosis. AB - Aspergilli are increasingly important infections in immunocompromised patients (ICP). The available antifungals often cause discrepancies in laboratory determination of MICs and a correlation in therapy. An effort was made to compare in vitro techniques for testing of antifungals, viz. polyenes, imidazoles, 5 fluorocytosine, amorolfine; and screened a phytoproduct- himachalol (a sesquiterpene alcohol) from Cedrus deodara (Roxb.) Loud against A. fumigatus clinical isolates (24) by macrobroth two-fold seal dilution (TFSD), microbroth microtitre (MT) and disc diffusion (DD) techniques using various broth/agar media at varying periods of incubation. The best activity in terms of geometric mean (GM) (GM.MIC < 0.39 microgrmas ml-1) was obtained with SCZ in the broth by both MT or TFSD technique followed by ECZ (GM.MIC 0.39 micrograms ml-1) and ITZ (GM.MIC 0.39-0.8 micrograms ml-1) in RPMI-1640. Overall RPMI-1640 was found to be the most suitable growth medium for testing of azoles or amorolfine, and YNB for polyene and 5-FC. MT technique was the most sensitive quantitative, reproducible, rapid and economical compared to other techniques. The treatment of Swiss mice with himachalol (200 mg kg-1, po) once a day, for 7 days, provided 60% protection concomitantly with increased MST (15 days) against invasive aspergillosis. A combination of himachalol (200 mgkg-1) plus SCZ (5 mgkg-1) showed better regimen in the therapy evidenced by enhanced survival (80%) of mice significantly (p < 0.001) with prolonged MST (> 15 days) compared to control. The treatments also reduced cfu (mean log10) burden of A. fumigatus from kidney. PMID- 9418376 TI - Effect of withafastuosin E on gastric mucosal offensive and defensive factors in rats. AB - Withafastuosin E (WE), a withanolide of Datura fastuosa, has been reported to possess anti-stress activity and augment prostaglandins. The present investigation has been undertaken to evaluate the anti-ulcer activity and its mechanism in various models of experimentally induced ulcers in rats. WE (20 mg/kg, po) reduced the incidence of ulcer and ulcer index significantly in rats. The drug also decreased volume of gastric secretion, acid and peptic output, though it did not affect mucin secretion or mucosal glycoprotein content in terms of total carbohydrate:protein ratio or gastric cell shedding (in terms of gastric juice DNA content) or cell replication (in terms of microgram DNA/mg of protein). The results suggest significant anti-ulcer activity of WE which may be due to its effect on decreasing the offensive acid-pepsin factors. PMID- 9418377 TI - Curative effects of mandur bhasma on liver and kidney of albino rats after induction of acute hepatitis by CCl(4). AB - Hepatocurative effects of mandur bhasma were studied in albino rats after induction of acute hepatitis by CCl4 liquid paraffin and CCl4 + liquid param. Recovery of the liver was studied with reference to histological architecture and differential counts of degenerated, recovering and recovered hepatocytes. Alterations in the kidney were also studied histologically. Hepatotoxins were given (s.c.) daily for 11 days. Mandur bhasma was given (po) for 7 days to normal, CCl4, liquid paraffin and CCl4 + liquid paraffin treated rats from day 12 to day 18. There were no spontaneous liver and kidney recoveries within a week after the cessation of the treatments of hepatotoxins. Mandur bhasma treatment showed conspicuous recoveries of liver and kidney within a week and total recoveries were noticed after two weeks. Biochemical alterations in lipid peroxidation, glucose-phosphatase and total proteins were studied during present work. The alterations in the histology and biochemical parameters of liver and kidney show hepatocurative potency of mandur bhasma. PMID- 9418379 TI - Cyclophosphamide-induced structural and biochemical changes in testis and epididymidis of rats. AB - Effect of cyclophosphamide administration (100 mg/kg body weight, ip, for 5 consecutive days) was studied on albino rat testis and epididymidis after 3 and 6 weeks of treatment. Cyclophosphamide decreased testis and cauda epididymidal weights, sperm count, motile and viable spermatozoa and increased percentage of abnormal spermatozoa. The biochemical changes observed in the testis include increase in acid and alkaline phosphatase activities and decrease in proteins and activity of lactate dehydrogenase. The levels of lipids and total cholesterol were not affected. In the epididymidis cyclophosphamide caused decrease in the tubular diameter and increase in its epithelial height. It is concluded that cyclophosphamide brings about structural and biochemical changes in the testis and epididymidis. PMID- 9418378 TI - Antiamoebic activity of 3,3'-fluro-4,4'-di-(pyrrolidine-2-ylidene amino)-diphenyl (liroldine), against experimentally infected intestinal and hepatic amoebiasis. AB - HL 707, Liroldine, a novel synthetic compound, was found effective against both extraintestinal and intestinal amoebiasis in animal models. Its activity against hepatic infection in golden hamsters is comparable with that of different derivatives of nitroimidazoles used for human treatment. Against intestinal amoebiasis in Wistar rats, the activity was superior to nitroimidazoles and chloroquine. Paramomycin was comparable and diloxanide furoate was marginally superior. The comparative in vitro and in vivo studies with standard marketed drugs and Liroldine indicate an excellent profile of the compound against experimental amoebiasis. LD50 of Liroldine determined in mice is 910 mg/kg x 1, po and 940 mg/kg x 1 ip). PMID- 9418380 TI - Effect of exposure of cigarette smoke on mechanical properties and thermal behaviour of rat skin and tendon. AB - Skin and tendon samples of male albino rat taken for analysis, on the 120th day of smoking showed that, compared to controls, the cigarette smoke exposed group showed an increase in tensile strength of both skin and tendon while extensibility of skin remained the same and that of the tendon increased. Thermal behaviour such as isometric tension and temperature at isometric tension increased in rat skin, while in tendon only isomeric tension increased. Shrinkage temperature of skin and tendon has showed no alteration in cigarette smoke exposed rats. PMID- 9418381 TI - Effect of experimental diabetes on the activities of hexokinase, glucose-6 phosphate dehydrogenase and catecholamines in rat erythrocytes of different ages. AB - Hexokinase and glucose-6-phosphate dehydrogenase were assayed in various circulating age fractions i.e., young, middle-aged and old red cell from control, diabetic and insulin-treated diabetic rats. An increase in the activity of hexokinase was observed in three age-wise separated fractions of red cells from diabetic animals in comparison to control. The activity of glucose-6-phosphate dehydrogenase on the other hand decreased in separated ageing fractions of diabetic red cells when compared to control. Reversal of these two enzymes were observed in insulin-treated diabetic rats. The levels of glycosylated haemoglobin and catecholamines were found to increase with ageing red cells in controls and also increased in red cells plasma. PMID- 9418382 TI - Effect of Gasex on pharmacokinetic profile of ciprofloxacin in healthy male volunteers. AB - Effect of Gasex, a herbomineral formulation on the bioavailability of ciprofloxacin was studied in six healthy male volunteers. On day one, single dose of ciprofloxacin (500 mg) was given at fasting state and the plasma Concentrations of ciprofloxacin were estimated after 1, 2 and 4 hr. On day 8 ciprofloxacin (500 mg) was given along with 2 tablets of Gasex and the plasma concentrations of ciprofloxacin estimated at the same time points. No significant difference in the plasma ciprofloxacin levels was found in these two treatments, indicating that the bioavailability of ciprofloxacin was not affected by simultaneous treatment with Gasex. PMID- 9418383 TI - Is age irrelevant? Perceptions of young and old adult eyewitnesses. AB - In Experiment 1, we videotaped elderly and younger adults (n = 69) reporting their memories of a crime video. The seniors were significantly less accurate than the younger adults. In Experiment 2, participants viewed the "testimony" videotapes and rated the elderly as less credible than the younger adults. In Experiment 3, participant-jurors (n = 302) evaluated transcribed testimonies provided by Experiment 1 participants. The ostensible age of the witnesses was manipulated. Thus, some participants read a senior's testimony believing it was provided by a younger adult and vice versa. Participants were apparently not biased by negative stereotypes of seniors' eyewitness capabilities. PMID- 9418384 TI - Recidivism rates among child molesters and rapists: a methodological analysis. AB - We address the high variability in sex offender recidivism rates by examining several of the critical methodological differences that underlie this variability. We used a dataset on 251 sex offenders (136 rapists and 115 child molesters) who were discharged over a 25-year period to examine changes in recidivism as a function of changes in dispositional definition of reoffense (e.g., arrest or conviction), changes in the domain of criminal offenses that are considered, and changes in the length of exposure time. The data indicate that: (a) both rapists and child molesters remain at risk to reoffend long after their discharge, in some cases 15-20 years after discharge; (b) there was a marked underestimation of recidivism when calculating a simple proportion (%) consisting of those who were known to have reoffended during the follow-up period, and (c) there was a marked underestimation of recidivism when the criterion was based on conviction or imprisonment. Forensic, clinical and policy implications of this high variability are discussed. PMID- 9418385 TI - Studies of repressed memory and the issue of legal validity. PMID- 9418386 TI - Nursing leadership: Christian character in chaotic times. PMID- 9418387 TI - The challenges of NCF ministry--an African perspective. PMID- 9418388 TI - Leadership development. PMID- 9418389 TI - Professionalism and ministry: are they compatible in nursing? PMID- 9418390 TI - Mentoring nurses into Christian leadership. PMID- 9418391 TI - Encouragement: possibilities unlimited. PMID- 9418392 TI - History in nursing--Lady McFarlane. PMID- 9418393 TI - Alternative site for intra-aortic balloon pump (IABP) catheter insertion. PMID- 9418394 TI - Clinical pathways and coronary artery bypass surgery. AB - Use of a multidisciplinary clinical pathway helps eliminate variations in patients' care. Organizing the care delivered each day of the patient's hospitalization may lead to fewer complications, a quicker recovery, and an earlier discharge. In today's healthcare arena, much attention is being focused on improving the quality of care and decreasing the need for acute care. Clinical pathways facilitate patients' outcomes and earlier discharge and thus reduce the cost of care. PMID- 9418395 TI - Successful use of a quality improvement team to reduce ventilator-associated pneumonia. PMID- 9418396 TI - Toxic epidermal necrolysis: a critical care challenge. PMID- 9418397 TI - Using the continuous quality improvement process to improve the care of patients after angioplasty. PMID- 9418398 TI - Using a critical care simulation laboratory to teach students. AB - Critical care nursing is a dynamic and continuous process involving knowledge, values and attitudes, and motor skills. When combined with other teaching methods, a critical care simulation laboratory provides learners with an effective means to achieve the cognitive, affective, and psychomotor competencies necessary for the complexities of clinical practice. PMID- 9418399 TI - Being led down the critical pathway: a perspective on the importance of care managers vs critical pathways for patients requiring prolonged mechanical ventilation. PMID- 9418400 TI - The quality connection: satisfaction of patients and their families. PMID- 9418401 TI - Trauma care strategies for changing economic forces. AB - Nurses have expertise in wellness, health promotion, delivery of acute care, and rehabilitation. As the venture into healthcare reform deepens, nurses must take a more proactive role in redirecting the delivery of trauma care in such a way that optimal provision of healthcare services is maintained while costs of providing care are reduced across the continuum of care. Efforts must focus on preventing traumatic injuries, restructuring healthcare delivery systems to meet the needs of patients with traumatic injuries, and reducing healthcare expenditures. Table 3 outlines strategies used by our facility to decrease cost without compromising patients' care. The current era is fraught with rapid changes that necessitate a creative, rational, and organized approach to making decisions about the delivery system for patient-focused care. Nurses are in an optimal position to develop and implement interdisciplinary, creative strategies that will maximize the delivery of trauma care services to the community. Each institution must evaluate the processes involved in its delivery of trauma care services. Strategies to contain costs must focus on processes implemented to achieve optimal outcomes of patients' care. The economic marketplace will evaluate care on the basis of outcome statistics and cost analysis. Thus, nurses must continue to be critical evaluators of nursing practice, always striving for the best healthcare delivery system possible during these turbulent economic times. PMID- 9418402 TI - Chronic disease management: an outpatient approach. AB - The current climate of managed care has sparked efforts to reduce costs in patient care. In many cases, this has resulted in more efficient methods of patient management: chronic disease management in an outpatient setting appears to be one such success story. For critical care nurses interested in working beyond the boundaries of a traditional ICU, chronic disease management clinics represent an alternative environment in which they may apply their skills. Nancy Brass-Mynderse, RN, MSN, CCRN, a clinical nurse specialist (CNS) with 18 years of experience in critical care, was instrumental in development of the Scripps Health Chronic Disease Clinic at Green Hospital of Scripps Clinic, San Diego, Calif. Brass-Mynderse currently supervises the operation of the clinic, along with Omana Kaliangara, RN, MSN, CFNP, a nurse practitioner. Brass-Mynderse received her bachelor's degree from the University of Arizona, Tucson, Ariz, and her master's degree from San Diego State University. She recently obtained her family nurse practitioner certificate from California State University, Dominguez Hills, Calif. Kaliangara received her bachelor's degree from San Jose State University, San Jose, Calif, and her nurse practitioner certificate from the University of California, San Francisco, Calif. After working in family medicine and a diabetic clinic, Kaliangara developed an interest in the management of chronic diseases. In an interview with CRITICAL CARE NURSE in September, Brass Mynderse and Kaliangara took time to discuss the development and operation of the clinic, and to recount some of their success stories. PMID- 9418403 TI - Direct arterial vs oscillometric monitoring of blood pressure: stop comparing and pick one. PMID- 9418405 TI - Reflections on "nursing at the cutting edge". PMID- 9418404 TI - Whose quality of life? PMID- 9418406 TI - The attitudes of nurses towards mentally ill people in a general hospital setting in Durban. AB - A quantitative survey was undertaken to determine the attitudes of nurses towards mentally ill people at King Edward VIII Hospital, large academic hospital in Durban. Data were collected by a questionnaire intended to measure attitudes according to cognitive, affective and behavioral components in a sample of 100 black nurses. The results of this study were analyzed through a statistical software package, the statgraphic version 5. The basic trend reflected in the findings found that nurses in general hospitals (90%) held negative attitudes towards mentally ill people and there were few nurses with positive attitudes. There were significant differences among the three different categories of nurses in only five of the items. Further data analysis was done by stepwise regression showing that religious affiliation and relationship to mentally ill people correlated significantly with the total attitude score. PMID- 9418407 TI - [Competency of newly qualified nurses following completion of the 4-year diploma program]. AB - The purpose of this study was to determine the competency of newly qualified nurses practitioners in a curative setting after completion of the 4 Year Diploma Course at Otto du Plessis Nursing College. The respondents evaluated their skills and abilities by completing a questionnaire which was developed by the research committee after consultation with the services. The need to assess competency comes from the desire to have a relevant curriculum enabling the newly qualified nurse to practice in a variety of health settings at all levels of care. The degree of competency related to practice in a curative setting was selected for particular attention because most newly qualified nurses in the target group were still employed in a curative milieu. Findings revealed various deficits or inadequacies in competency levels and recommendations were made to adapt the curriculum. PMID- 9418408 TI - Blood glucose control and compliance of diabetic children. AB - AIM OF STUDY: Non-compliance is an important factor hindering good control in diabetics. The aim of this study was to identify areas of poor compliance with the diabetes management regimen in the children attending our clinic. DESIGN: A questionnaire was administered to 57 patients who attend the Paediatric Diabetes Clinic. It was designed to elicit socio-demographic data and information about the diabetic regimen. Prior to the administration of the questionnaire, patients were classified as being well, satisfactorily or poorly controlled, based on their average glycosylated Haemoglobin results over the past year. RESULTS: All the patients used home blood glucose monitoring (HBGM)-79% of the poorly controlled children tested twice daily or less whereas 53% of the well controlled children tested three times or more daily. The timing of injections was frequently incorrect. 42% of all patients had been admitted to hospital after diagnosis and more than 60% of them never tested their blood glucose in relation to exercise. The patients' knowledge about their disease was generally good. The mean age of the poorly controlled group was almost 19 months older than that of the well controlled group. Poorly controlled children had also had diabetes for longer and they lived significantly further from the hospital. A higher percentage of poorly controlled patients were in charge of their own treatment while those in the well controlled group were less reliant on doctors for insulin dose adjustments. PMID- 9418409 TI - [The effect of constant baroque music on premature infants]. PMID- 9418410 TI - An assessment of the quality of care given to, and hygiene on patients at a teaching hospital in Namibia. AB - An assessment was done during 1991 to evaluate the quality of care with regard to mouth hygiene rendered to patients in a teaching hospital in Namibia. The sample was drawn from nine wards. By means of a type of quota sampling, the patients were categorised as dependent, interdependent or independent. The nursing process was used as a framework for the study. From the assessment it became evident that no policies existed with regard to oral hygiene. Planning was not in every case based on assessment, and it seemed that when planning(s) were done, it was not always implemented. Record keeping was the aspect most poorly attended to. PMID- 9418411 TI - The measurement of quality of care in public sector psychiatric services based on consumer expectations. AB - In this study the expectations of consumers of public sector psychiatric care in South Africa were identified, and formulated in the form of 13 standards, each with a set of criteria. During this phase input from the literature was incorporated, and expectations were validated with different groups of consumers, so that rural/urban, ethnicity and regional differences were taken into account. Based on the comprehensive set of standards and criteria, four instruments were developed to measure attainment of these standards. These included a questionnaire to consumers and one to the Director of Mental Health. It also included two schedules to be filled in by observers during site visits to hospital units and clinics. The observer teams included community members and consumers. The content validity of the instruments was established by setting out the items measuring each criterium, and validating that with a group of experts. The instruments were then tested in one province. The inter-rater reliability of the site visit schedules was calculated as 0.94, and the coding of the Director questionnaire by different coders was also tested. The average performance on all criteria was calculated, using items from all four data collection instruments. In the process items were revised, coding instructions developed, and criteria adjusted. PMID- 9418412 TI - [Development of leadership in a nursing college]. AB - This study is conducted within the context of nursing colleges and focuses on the empowerment of educational staff in a nursing college to facilitate excellence in leadership practice. A qualitative exploratory and descriptive study was conducted in different phases: the views and expectations of educational staff as leaders, as well as the views and expectations of the fourth year students as followers and potential professional leaders were explored and described by means of focus group interviews. A provisional conceptual framework for leadership development in a nursing college was compiled, exposed to a literature control and a final conceptual framework was refined, together with guidelines for leadership development in a nursing college. This article focuses on the final conceptual framework as well as the programme for leadership development in a nursing college. It is recommended that this programme be implemented and exposed to purposeful evaluation. PMID- 9418413 TI - Health education: a baby show as an evaluation tool. PMID- 9418414 TI - Professional nurses knowledge and understanding of AIDS/HIV infection. AB - This study examined professional nurses' knowledge and understanding of AIDS/HIV infection. The sample was formed by 53 professional nurses; 27 being those that have done the AIDS counselling course (trained counsellors) and 26 who have not done the course. This sample was randomly chosen from a population of 192 professional nurses representing all departments. Questionnaires were sent to the participants and focus group interviews formed by a group of 21 participants, were done to elicit any information that may not have been obtained through the use of a questionnaire. The results showed good knowledge of general information which included the mode of spread of infection. Lack of knowledge in identification of high risk groups, symptoms, diagnostic tests and use of universal precautions in specific areas was identified. PMID- 9418415 TI - [Management competency of persons registered as disaster nurses in the Pretorian Civil Defense]. AB - The essential management role of the disaster nurse during disaster action was outlined, researched and described. Her competency to execute effectively disaster relief tasks before, during and after a disaster occurring outside a hospital, was studied. Management tasks were identified which nurses should have mastered regarding disaster situations occurring outside hospital boundaries. Research data were gathered by means of a questionnaire on the biographic detail of disaster nurses registered with Civil Defence in Pretoria, in order to recommend a course specifically aimed at fulfilling their requirements. The research project identified requirements of the disaster nurse for appropriate further training, practise and guidance regarding the identified management tasks. It became evident that training is required in most of the tasks, and a training course for nurses in disaster management was designed. PMID- 9418416 TI - Research as intervention within community mental health. AB - This paper raises some issues for discussion and debate concerning the nature of research within a mental health setting. Research, no matter what form it takes, is always an intervention. Sensitivity to various concerns surrounding research is required of the mental health worker. Participatory research, which is seen as empowering participants, has become popular in mental health programmes. Attention needs to be paid, however, to the meaning of participation and the process of decision-making. Researchers are often uncomfortable with shedding their "objective informer" stance and adopting a position that requires social action. Some of this has to do with the epistemological view taken by the researcher. This paper suggests that the knowledge produced in research is a social construction created in interaction between the researcher and participants and has a multiplicity of potential meanings. The instrumental, conceptual and persuasive uses of research are discussed, and two intervention type research procedures (needs assessment and evaluation) are critically reviewed. It is concluded that research, as with all other interventions, should be carefully planned, implemented, monitored and evaluated. PMID- 9418417 TI - [Operationalization of a model for the development of clinical nursing standards]. AB - One of the requirements for a nursing model in any field of nursing is its practical implementation. The purpose of this article is to describe how the model for the development of nursing standards in a nursing service was implemented and evaluated. An exploratory and descriptive research design was followed, with a simple pilot study during which 9 nurses were requested to give their view on the model. The model was sucessfully implemented in the nursing service of an academic hospital. Evaluation revealed that the implementation facilitated quality nursing, the resocialising of values and the professional and personal growth of nurses in that nursing service. PMID- 9418418 TI - The traditional bio-medical dichotomy in search of a common ground. AB - Differences between bio-medical and traditional systems are regarded as irreconcilable by many, including members of the medical, nursing and allied professions. This article explores those areas in which emphasis on difference has resulted in an exclusion of the common basis from which culturally determined areas of human and health behaviours are derived. As long as difference is emphasized to the exclusion of attempts to find a common ground, the dichotomy between traditional and bio-medicine will remain. PMID- 9418419 TI - [A program to facilitate quality of work-life of psychiatric nurses]. AB - The aim of this study was to describe guidelines for a personal and professional development programme to facilitate the quality of work-life experienced by psychiatric nurses in a hospital. An explorative and descriptive research design with a qualitative research orientation was employed. The study was divided into three phases. In phase one the needs, desires and expectations of psychiatric nursing in a hospital nursing service were explored and described. In phase two the factors in a nursing service which influence the quality work-life of nurses, were explored and described. Consequently, the last phase of the study was conducted, being inferred from data of phases one and two which lead to the conceptual framework upon which the guidelines and programme are based. A personal and professional development programme for psychiatric nurses to facilitate quality of work-life experienced, consisting of three parts, was described to enclose aspects of the psychiatric nurse's internal and external environments, as well as patterns of interaction between the internal and external environments. PMID- 9418420 TI - An investigation into the effects of stress on the dynamics of selected Xhosa families. PMID- 9418421 TI - Backstreet abortion: women's experiences. AB - AIM: This was a descriptive study aimed at exploring the personal experiences of women who induce abortion and the circumstances surrounding induced abortion. METHODS: The study was conducted in six public hospitals in four different provinces: Baragwanath (Gauteng), Groote Schuur and Tygerberg (Western Cape), King Edward and R.K. Khan (Kwa-Zulu/Natal) and Livingstone (Eastern Cape). In depth interviews were conducted with 25 African, Indian and Coloured women admitted to the hospitals following backstreet abortions. The study gave women the opportunity to "speak for themselves" about "why" and "how" and the context in which the unsafe induced abortions occurred. RESULTS: The findings show that a host of factors were important in the circumstances leading to unwanted pregnancy and induced abortion: socio-economic, cultural, psychological and societal. Disempowerment in relationships combined with financial pressures constituted the background as to why women felt forced to terminate their pregnancies. The perceived need for termination was found to over-ride all other considerations, including religious ones. The ways in which women attempted to procure abortion, both through legal and illegal routes, are presented. Wider social and legal discourses an abortion were found to be an important factor in how women experienced their situation. PMID- 9418423 TI - Creating partnerships for delivering community services. Involving nursing students in American Red Cross activities. PMID- 9418422 TI - Culture bound syndromes in a group of Xhosa with psychiatric disorders. AB - The article describes the culture bound syndromes found amongst a sample of 40 psychiatric patients seen at Umzimkulu Hospital serving mainly Xhosa speaking people. A CBS could be identified in 15 of these patients. CBS similar to four of the six syndromes found in these patients were described in DSM IV, while two were not. The CBS were compared to the Axis I diagnoses, and the different syndromes described. PMID- 9418424 TI - Finding new and better ways to touch--keeping ahead of healthcare changes. PMID- 9418425 TI - Joint Commission on Accreditation of Healthcare Organizations' infection control requirements: fact or fiction? PMID- 9418426 TI - Nursing strategies for preventing home health aide abuse. AB - One of home care's most important resources is the home health aide. Home care nurses play a critical role in preventing abuse of home health aides and identifying violence-prone environments. A prevention strategy that nurses can use to identify and prevent abuse of both patients and aides is presented using an Assessment, Communication, Education, and Supervision model. PMID- 9418427 TI - The home health aide as a member of the home healthcare team. AB - Individuals of all ages are discharged from acute-care facilities with a variety of medical and nursing diagnoses. Many of these individuals require home health aides services in addition to their skilled care. In the changing healthcare environment of the 1990s, home healthcare professionals are being challenged to provide effective, skilled care that is the most cost-effective and appropriate for patients. It is important that all professional home healthcare personnel work as a team with home health aides to accomplish these goals. The home health aide is an important member of the home care team. PMID- 9418428 TI - Home care for the patient receiving mechanical ventilation. PMID- 9418429 TI - Depression in elders with dementia: implications for home healthcare practice. AB - In addition to severe cognitive decline, depressive symptoms occur in almost 50% of patients with dementia. Depressive symptoms can be treated successfully, thereby reducing clinical manifestations of coexisting dementia and improving quality of life for these patients and their families. Practical assessment, interventions, and referral guidelines are presented for home healthcare nurse generalists whose caseloads include elders with both cognitive decline and depressive symptoms. PMID- 9418430 TI - Fifteen common errors to avoid when completing a skilled home care referral. PMID- 9418431 TI - Ask home healthcare nurse. PMID- 9418432 TI - Environmental assessment. AB - The significance, standard elements, components, and documentation of an environmental assessment in home care are discussed. This assessment is delineated within Maslow's Hierarchy of Needs and from a functional perspective. An Environmental Assessment Form that can be used as a documentation tool is included. PMID- 9418433 TI - Informational needs of caregivers of terminal patients in a rural state. AB - Home care of a terminally ill family member is stressful, especially in rural areas. This qualitative study sought to determine informational needs of rural caregivers and how that information is obtained. Although most caregivers stated satisfaction with available information, mostly obtained from physicians and nurses, their behavior belied their satisfaction. Assertive and self-reliant, they used informal communications rather than written information to meet most of their needs. Approaches home care nurses can use to help caregivers obtain important information are presented. PMID- 9418434 TI - We must look at our similarities, not our differences. PMID- 9418435 TI - Ethics and cultural sensitivity: co-principal conduct for effective human relation. PMID- 9418436 TI - Ethnicity and morality in a multicultural world. AB - The choice of moral acts is governed by the unique ethnic and/or cultural meaning and values ascribed to by the individual. In pursuing an "acceptable" moral choice, the process of analysis is further directed by the ethical theories which are molded by time and tempered by the current context to which the action is undertaken. An OPTIMIN MODEL OF ETHICAL-DECISION MAKING PARADIGM is proposed, and health care policy on cultural diversity is adduced. PMID- 9418437 TI - Co-family sleeping: strange bedfellows or culturally acceptable behavior? AB - Community health, psychiatric, and pediatric nurses who work with young children and their families often assess the family's sleep and rest patterns, especially in relation to children's sleep disturbances. Nurses have traditionally taken a rigid approach on this issue that excludes culturally diverse options. The author uses examples from her own nursing education and from cross-cultural research to demonstrate the cultural bias favoring separation of child from parent. The article explores options for broadening nurses' assessment of family sleeping arrangements and designing interventions that take into consideration culturally diverse values and habits. Implications for nursing practice include greater trust-building and mutual learning. PMID- 9418438 TI - Asian/Pacific Islander American nurses workforce: issues and challenges for the 21st century. AB - The trend of high incidence of lung cancer, cardiovascular disease, Hepatitis B, and tuberculosis among the Asian/Pacific Islander Americans (APIAs) will significantly affect the increased need for culturally competent care for this particular ethnic group. There is a need to increase the numbers of Asian/Pacific Islander American nurses in the future workforce to meet the health needs of this heterogenous and diversified population. Current workforce data on the Asian/Pacific Islander American nurses is misleading, since data collection aggregates all APIAs into a single category, with disregard to the various subgroupings of this large ethnic group. PMID- 9418439 TI - What are some African American working women's expressed experiences of role conflict? AB - The purpose of this study is to explore and describe the experiences of African American working women who attended a six week intervention group to decrease role conflict and enhance psychological well-being. The sample consisted of nine African American working women ranging in age from 29 to 55. Participants reported role conflict with their professional roles, which was further confounded by race. Areas of role conflict included: (a) scrutinization, (b) de legitimization of the professional role, and (c) horizontal abuse. It was concluded that African American working women experienced an additional element of role conflict that is related to their ascribed role rather than their achieved role. PMID- 9418440 TI - Folk medicine and health beliefs: an Appalachian perspective. AB - This article examines Appalachian folk medicine and considers the importance of utilizing an integrative theory, the Health Belief Model, to understand the use of alternative medicine in the United States. The author examines the need for social workers and other health care providers to further assess the roles of folk medicine in an Appalachian client population. The author creates linkages among the limited writings in the literature regarding folk medical practices of Appalachians and also draws examples of folk medicine usage from qualitative research and professional intervention with this client group. The author focuses on how mainstream health care professionals may assess the role of folk medicine in the lives of their clients by approaching folk medicine as a focal and culturally-imbedded component of their clients' overall health care. Moreover, the author addresses the need for health care professionals to become not only aware of folk medical practices, but to act as advocates for culturally competent health care within the larger health care delivery system which largely overlooks or downplays the significance of folk medicine. In an age of expensive and constricted mainstream health care services and the implementation of managed care, the author examines how folk medical practices factor into primary health care. PMID- 9418441 TI - Library access is one of the most important issues for nurses in today's knowledge-conscious health service. PMID- 9418442 TI - Practice makes perfect. PMID- 9418443 TI - Watch with mother. PMID- 9418444 TI - Bug-eyed monster. PMID- 9418445 TI - Check them out. PMID- 9418446 TI - Empty vessels. PMID- 9418447 TI - Magnet hospitals: what's the attraction? PMID- 9418448 TI - Mixed bag. PMID- 9418449 TI - Doing the right thing. PMID- 9418450 TI - Developing a research culture in education and practice. AB - In 1996, the English National Board held discussions with five focus groups of nursing and midwifery lectures with an interest in or responsibility for the development of research. The purpose of the groups was to explore the experiences of lecturers in colleges which had recently integrated with higher education institutions. The focus groups were asked particularly about their experiences in developing research expertise to meet their professional and academic needs, the demands of the research assessment exercise (RAE) and the NHS research and development (R&D) agenda. PMID- 9418451 TI - A new account of sundown syndrome. AB - In this study, the authors tested the hypothesis that residents requiring nursing care rather than residential care in a dual registered nursing and residential home would experience increased levels of confusion and agitation as the evening drew in. This article describes how the hypothesis was examined and the action that was taken as a result. PMID- 9418452 TI - The quality of district nursing care for dying patients. AB - During a project to explore the quality of district nursing care provided to patients with and without cancer, six district nurses in one community trust were interviewed. They acknowledged that the quality of their terminal care was dependent on whether patients were dying from cancer or from non-cancer diseases. This article describes how the project was undertaken, the methods, the results and the implications for district nursing. PMID- 9418453 TI - Life-long education: problem-based learning. Part 1. AB - In the first of two articles, the authors describe how nurses in the urology department at Battle Hospital overcame several obstacles and developed a new learning module which could eventually be used by all nurses whatever their experience. The second article will discuss how the module has developed and will be published shortly. PMID- 9418454 TI - The foundations of allergy. AB - This article discusses the immune system in relation to allergy. The importance of the role of the nurse in identifying and referring patients who have the most common allergic diseases is also emphasised. PMID- 9418455 TI - Advancing practice in the care of older people. PMID- 9418456 TI - Read all about it. PMID- 9418457 TI - No easy answers. PMID- 9418458 TI - Prejudice and paranoia. PMID- 9418460 TI - Relative rights. PMID- 9418459 TI - First class nursing without degrees. PMID- 9418461 TI - Humanising a hidden service. Interview by Ian McMillan. PMID- 9418462 TI - All graduate status. PMID- 9418463 TI - Get in the right lane. PMID- 9418464 TI - The pain attraction. PMID- 9418465 TI - The impact of information. PMID- 9418466 TI - Improving palliative care: audit and standards. AB - This report is based on an occasional paper prepared for the National Council for Hospice and Specialist Palliative Care Services. The paper acknowledges that although quality is already central to the palliative care approach, this is not usually in the context of multidisciplinary and collaborative audit. The document offers guidance on how to translate the principles of quality into an action programme for all professionals involved in palliative and hospice care. PMID- 9418468 TI - Childhood constipation and the incidence of hospitalisation. PMID- 9418467 TI - Evaluating the quality of after death care. AB - Nursing care rarely ceases following a patient's death. The project described in this article concerns the quality of nursing care delivered to patients following their death. The author concludes that the quality of care depends in large part on whether staff received training in the care of dying patients. PMID- 9418469 TI - Conservative treatment of urinary incontinence. PMID- 9418470 TI - Care of people with learning disabilities in hospital. PMID- 9418471 TI - Neurological observations--2. Pupil response. PMID- 9418472 TI - Nutrition and eating disorders. PMID- 9418473 TI - It is a cop-out to regard nurses who harm patients as simply evil or mad. PMID- 9418474 TI - Burnt into our memory. PMID- 9418475 TI - Throw down the gauntlet. PMID- 9418477 TI - Listening brief. PMID- 9418476 TI - Troubled union. PMID- 9418478 TI - When profits come first. PMID- 9418479 TI - Rights from the start. PMID- 9418480 TI - Exclusion of care. PMID- 9418481 TI - Care and secrets. PMID- 9418482 TI - Tales of deprivation. PMID- 9418484 TI - Nurses who kill--dangerous assumptions. PMID- 9418483 TI - Nurses who kill--counting the dead. PMID- 9418485 TI - Nurses who kills--damned if you do.... PMID- 9418486 TI - Nursing homes--a question of inspection. PMID- 9418487 TI - Hot potatoes--lashings of double standards. PMID- 9418488 TI - A practice-centred model of clinical supervision. AB - The practice-centred model of supervision (PCMS) has been developed in response to the UK nursing professions' imperative to establish efficient and effective systems of clinical supervision. The model has been designed in consultation with clinicians and managers in over 30 NHS trusts. PCMS has recently been the subject of a randomised controlled trial in one NHS trust. This article outlines the model. PMID- 9418489 TI - Home for people with challenging behaviours. AB - This paper describes the first three years in the life of a group home for two men with learning disabilities and seriously challenging behaviour. After initial optimism, problems were soon encountered, resulting in high levels of staff turnover and sickness associated with an increase in the residents' challenging behaviour. A number of changes were introduced, aimed at both staff and residents, which seem to have improved staff well-being and reduced challenging behaviour. The importance of a clear structure and effective support system, combined with focused intervention for residents, is emphasised as important for success. PMID- 9418491 TI - The route to true autonomous practice for midwives. PMID- 9418490 TI - The immune system, Part 2: Myasthenia gravis. PMID- 9418492 TI - Wound care--You'll never work alone. PMID- 9418493 TI - Student nurses in the wound care team. PMID- 9418494 TI - Wound care--identity crisis. PMID- 9418495 TI - Know how--alternating pressure relief. PMID- 9418496 TI - Wound care--healing messengers. PMID- 9418497 TI - Wound care--recovery on a plate. PMID- 9418498 TI - Wound care--less pain, more gain. PMID- 9418499 TI - Community nurses have demonstrated commitment. PMID- 9418500 TI - Flashback to my own baby killer nightmare. PMID- 9418501 TI - All cut up about men in leather. PMID- 9418502 TI - Not in with the in-crowd. PMID- 9418503 TI - Will ye no come back again? PMID- 9418504 TI - Our obsession with resuscitation. PMID- 9418505 TI - Could do better. PMID- 9418506 TI - A garden of the imagination. PMID- 9418508 TI - Neurological observations--3 Limb responses. PMID- 9418507 TI - An essential and ancient oil. PMID- 9418509 TI - Is curiosity killing nursing? PMID- 9418510 TI - Time of transition. PMID- 9418511 TI - Halfway home. PMID- 9418513 TI - Acting up. Interview by Richard Morris. PMID- 9418512 TI - Swingers. Interview by Martin Vousden. PMID- 9418514 TI - What is real nursing? PMID- 9418515 TI - Breast-feeding support in a neonatal surgical unit. AB - The focus of this article is breast-feeding support for mothers of infants that have to be nursed in hospital. It examines previous studies and considers them alongside the findings of an anonymous customer-satisfaction survey. Indications are that a long-term commitment of resources is required if breast-feeding support for this high-risk population is to be improved. PMID- 9418516 TI - Behaviour modification in hyperactive children. AB - Behaviour problems in pre-school children can be significantly improved by professionals experienced in child behaviour modification techniques. Reported disobedience, temper tantrums and poor concentration in hyperactive children aged three to three-and-a-half years improved significantly in a treatment group, following eight home visits by research health visitors working in a child and family guidance service. PMID- 9418517 TI - Diversion schemes: an alternative to court. PMID- 9418518 TI - Influencing the health agenda. PMID- 9418519 TI - [Report of the 12th General Members' Meeting of the Society of Oncologic Nurses]. PMID- 9418520 TI - [Report Symposium: "Dilemmas for nurses concerning palliative care and euthanasia"]. PMID- 9418521 TI - [Work Group palliative care--debate theme palliative care]. PMID- 9418522 TI - [A comparative study between 3 venous access systems with various types of catheters]. PMID- 9418523 TI - [Report Pain Symposium: 'From research to clinical practice: overcoming the pain problem']. PMID- 9418524 TI - [Working on quality: nothing new, but in another guise]. PMID- 9418525 TI - [What do you mean no anamnestic interview in the cytostatics outpatient clinic?]. PMID- 9418526 TI - [Report action on fatigue and its consequences]. PMID- 9418527 TI - [Nurse's guideline: a guide in complicated care]. PMID- 9418528 TI - [The pills are to blame!]. PMID- 9418529 TI - [Experiences from St. Christophers Hospice in London]. PMID- 9418530 TI - [Formation Nurses' Work Group Head Neck Oncology (VWHHO)]. PMID- 9418532 TI - [Annual report of the Association of Oncologic Nurses, 1996]. PMID- 9418531 TI - [KNMI (Royal Dutch Meteorologic Institute) start with predictions on the sun's power]. PMID- 9418533 TI - [The myth of home care. The matter-of-course of transfer of care to caregivers and family of cancer patients]. PMID- 9418534 TI - [Ethical dilemmas in oncologic nursing]. PMID- 9418535 TI - [Wiser in the future, clever in the past]. PMID- 9418536 TI - ['Encourage patients to tell their story']. PMID- 9418537 TI - [The nurse in palliative care]. PMID- 9418538 TI - [Developments in palliative care on ward b2 West; internal medicine, oncology and AIDS]. PMID- 9418539 TI - [Palliative care in The Netherlands; which direction?]. PMID- 9418540 TI - [Health promotion from an ecological viewpoint]. PMID- 9418541 TI - [Autonomy and concern for excellent care]. PMID- 9418542 TI - [Health promotion at the time of discharge from the hospital]. PMID- 9418543 TI - [Should cytostatic agents be administered at home?]. PMID- 9418544 TI - [Quality of life in young adults who had cancer at an early age]. PMID- 9418545 TI - [Hereditary breast and/or ovarian cancer: consequences for family relations]. PMID- 9418547 TI - [Report Society of Pediatric Oncology (SIOP), Vienna]. PMID- 9418546 TI - [Oncologic nursing on the map--results of a survey among congress visitors]. PMID- 9418548 TI - [Quality of life: 1525 reactions of cancer patients]. PMID- 9418549 TI - [Police always ready for a noncommitted talk. Interview by Anne Vesterdal]. PMID- 9418550 TI - [Medical establishment complains about Falck]. PMID- 9418551 TI - [Vaccinations: in the service of health. Interview by Dorthe Nerving]. PMID- 9418552 TI - [The Danish immunization program]. PMID- 9418553 TI - [Taking the offensive in health debates]. PMID- 9418554 TI - [Psychological worjing environment--on-the-spot assistance. Interview by Niels Seeberg]. PMID- 9418555 TI - [New special education for nurses]. PMID- 9418557 TI - [Handling of drugs--a nuisance with use of blister packaging]. PMID- 9418556 TI - [The good with the bad]. PMID- 9418558 TI - [Handling of drugs--pain, irritation and distaste. Interview by Grethe Kjaergaard]. PMID- 9418559 TI - [Health policy's values and goals--introduction to a debate]. PMID- 9418560 TI - [Adoption can be difficult]. PMID- 9418561 TI - [Adoption--overlooked by health system. Interview by Kari Anne Aase]. PMID- 9418562 TI - [AF breakup (Joint Organization of Academicians)--ex-nurse to be next leader of Academicians. Interview by Kjell Arne Bakke]. PMID- 9418563 TI - [Viewpoint from the community physician. Interview by Marit Fonn]. PMID- 9418564 TI - [An ailing soul in a healthy body]. PMID- 9418565 TI - [Closeup: Maj-Len Sundin, director, Aker Hospital, Oslo. Stamina at the top. Interview by Marie Dyren]. PMID- 9418566 TI - [Work and security--from sofa to job. Interview by Marit Fonn]. PMID- 9418567 TI - [My workplace: intensive Care Unit, Notodden Hospital. Interview by Erik Dale]. PMID- 9418568 TI - [From bygone days: romantic figures (Florence Nightingale)]. PMID- 9418569 TI - [Care for the aged--steadily fewer institutional places]. PMID- 9418570 TI - [Slipped disk--one out of 3 has ulcers. Interview by Kjell Arne Bakke]. PMID- 9418571 TI - [Psychiatry: violence--a taboo problem in the working environment. Interview by Erik Dale]. PMID- 9418572 TI - [Depression--when the dark of winter arrives]. PMID- 9418573 TI - [World AIDS Day--children in a world with AIDS]. PMID- 9418574 TI - [Adverse effects of formalin, compensation matters and the NSF (Norwegian Nurses' Association)]. PMID- 9418576 TI - [Education--a symbolic encounter with nursing practice]. PMID- 9418575 TI - [Pregnancy--extreme pregnancy nausea gets little attention]. PMID- 9418577 TI - [Grief work--youths need support in grief also]. PMID- 9418578 TI - ["The most important thing is to be liked by doctors"]. PMID- 9418580 TI - [From hours to consultations? Honest competition in home care]. PMID- 9418579 TI - [Reaction to the Den Treek memorandum on faculty nursing practice 'Learning is subordinate to working in clinical practice']. PMID- 9418581 TI - [Nurse specialist oncology can coordinate intra- and extramural care. The child with cancer at home: whose care?]. PMID- 9418582 TI - [Living environment, abilities and function as central elements. Diagnosis in nursing: field of attention]. PMID- 9418583 TI - [Implications for nursing care. Insight into illness in patients with schizophrenia]. PMID- 9418584 TI - [Nurses evaluate each other's work. Visit with a tail]. PMID- 9418585 TI - [Biennial professional fair for health care. Medica '97 in the age of care and commerce]. PMID- 9418586 TI - ['Combination function for nursing instructors'. Learning in clinical practice with the clinical nursing instructor. Interview by Petra Q. Nijdam and Patrick Arink]. PMID- 9418587 TI - [Team work between family physician, clinical assistant and nurse practitioner: dilemmas and hot potatoes. The nurse practitioner in a triangular situation]. PMID- 9418588 TI - [Diagnosis in nursing: history taking and other data]. PMID- 9418589 TI - [Training funds no longer transferred to health care facilities. The Institute Clinical Education. Interview by Tonny van de Pasch]. PMID- 9418590 TI - [Current treatment policy gives considerably better results--CVA and its sequelae]. PMID- 9418591 TI - [The rehabilitation program often shows a gap--psychosocial consequences of CVA]. PMID- 9418592 TI - [Heart Foundation's campaign concerning CVA--'stroke, today's biggest headache'. Interview by Patrick Arink]. PMID- 9418593 TI - [Rehabilitation following hospital discharge. Nursing problems of CVA-patients and caregivers]. PMID- 9418594 TI - [Hospital nurses can give caregivers and relatives oriented support. Care for the caregiver of a CVA-patient. Interview by Tonny van der Pasch]. PMID- 9418595 TI - [Coordinating task reserved for transmurally- and extramurally active nurses. The long-term impact of a stroke]. PMID- 9418596 TI - [Quality of care improves through team work--caregivers form chain of care for CVA-patients]. PMID- 9418597 TI - [Searching for the person behind the patient. Nursing with distinction of the individual]. PMID- 9418598 TI - [Social context demands a closer orientation of nurses. 4 scenarios for the future of nursing]. PMID- 9418599 TI - [Problems in home care not yet coming to an end. Commotion in home care]. PMID- 9418600 TI - [CFO rings alarm bell about threatened firing of clinical instructors]. PMID- 9418601 TI - [2d European Nursing Congress goes into chronic disease care. 'Welcome to Amsterdam']. PMID- 9418602 TI - [Limit setting as an important task for nurses in ambulatory drug abuse treatment. More than handing out cups of methadone]. PMID- 9418603 TI - [Data-sets as flexible tool in nursing plans. Care of patients with HIV/AIDS benefited by clearness]. PMID- 9418604 TI - [Faculty-practice nurse]. PMID- 9418606 TI - [The Nursing Scientific Council builds a bridge between theory and practice. The VWR in the nursing field. Interview by Tonny van de Pasch]. PMID- 9418605 TI - [2 orientations in nursing]. PMID- 9418607 TI - [Decubitus: our problem]. PMID- 9418608 TI - [Guidelines for decubitus prevention are not well enough known. Massage does not help]. PMID- 9418609 TI - [No standard fits every patient. Time interval in alternating position]. PMID- 9418610 TI - [Steering Group has big plans--national approach to decubitus]. PMID- 9418611 TI - [The faculty-practice nurse]. PMID- 9418612 TI - [Consensus is developing within nursing. CBO/VWR guideline 'fluid balance' introduced in the VU-Hospital (Free University). Central Guidance Organization for Peer Review/Nursing Scientific Council]. PMID- 9418613 TI - [More legal actions against caregivers]. PMID- 9418614 TI - Management of open fractures and open wounds of joints. 1968. PMID- 9418615 TI - The Coventry Award. The value of preoperative aspiration before total knee revision. AB - The value of routine aspiration of a symptomatic total knee replacement before reoperation was evaluated. The study group consisted of a consecutive series of 69 knees in 67 patients in which preoperative aspiration was performed. All aspirations were performed on an outpatient basis in a clinic setting. Local anesthetics and saline washings were not used. Twenty knees were determined to be infected and 49 knees were not infected. Preoperative aspiration had an overall sensitivity of 55%, specificity of 96%, accuracy of 84%, positive predictive value of 85%, and negative predictive value of 84%. Sixteen patients were taking antibiotics at the time of referral including 12 of 20 (60%) who had infected knees. Seven of these 12 (58%) had no growth on their initial knee aspiration. Four of these had their knees reaspirated at a later date because of a high index of suspicion for infection and the subsequent aspiration revealed the infecting organism in all four cases. Two of the remaining three patients had signs of sepsis develop and reaspiration was not performed because immediate reoperation was indicated clinically. The initial aspiration on the third patient was performed after antibiotic therapy was discontinued for 4 weeks and a repeat aspiration was not deemed necessary. When the results of the reaspirations are included, the overall aspiration results improved to a sensitivity of 75%, specificity of 96%, and accuracy of 90%. The results of the study support the use of routine preoperative aspiration before total knee revision. Previous antibiotic use increases the risk of a false negative result, and reaspiration at a later date can be expected to significantly improve the value of this test in such cases. PMID- 9418616 TI - The Ranawat Award. Histology of nine structural bone grafts used in total knee arthroplasty. AB - The cross section radiographs and histology of nine bone grafts were examined to determine whether grafts are durable enough to support a total knee implant when the load is shared by host bone, graft bone, and a stemmed component. All cases had cemented total knee arthroplasties with stemmed components adjacent to bulk grafts. The cases included autografts and allografts, which had been in situ for an average of 41 months (range, 20-62 months). Seven of the grafts were retrieved postmortem from three patients (four knees), and two were retrieved at revision surgery from one patient. The allografts all were intact, but had not revascularized. The autografts were viable bone. New bone was being laid down on the dead graft bone at the periphery of the allografts. No change in the bone to cement interface, no graft collapse, no development of radiolucent lines, and no component loosening occurred in these cases. The promising clinical results of bone grafts in total knee arthroplasties were confirmed by the examination of these grafts at the cellular level. Using stemmed components in bone grafted knee reconstructions may have increased graft durability and protected the grafts from fatigue failure. PMID- 9418617 TI - The Insall Award. Total knee replacement with posterior cruciate ligament retention in rheumatoid arthritis. Problems and complications. AB - A series of patients with rheumatoid arthritis underwent total knee replacement with posterior cruciate ligament retention and was observed a minimum of 6 years and a mean of 8.2 years. A group of patients with osteoarthritis with an identical prosthesis and a group of patients with rheumatoid arthritis with a posterior stabilized implant served as controls. In the rheumatoid arthritis group with posterior cruciate ligament retention, there was an increased incidence in posterior instability and recurvatum deformity, resulting in an increased revision rate. Those patients undergoing revision for instability had a higher incidence of recurrent synovitis, and at revision the posterior cruciate ligament was grossly absent with a Grade 1 synovial reaction. In patients with rheumatoid arthritis undergoing total knee replacement, a posterior stabilized prosthesis rather than a posterior cruciate ligament sparing prosthesis should be used. PMID- 9418618 TI - 10- to 20-year followup of total knee arthroplasty for valgus deformities. AB - One hundred eight knees in 83 patients with a valgus alignment of greater than 10 degrees underwent total joint replacement performed by a single surgeon using the same technique for ligament balancing, which involved releasing the lateral retinaculum and iliotibial band, followed when necessary by detaching the lateral collateral ligament and popliteus tendon from the femur. Sixty knees in 46 patients had followup of at least 10 years and were the focus of study. At an average followup of 14.1 years, the mean Knee Society knee score was 88.7 and the mean functional score was 69.2. Postoperative knee alignment averaged 4.5 degrees with 75% of the knees corrected to between 2 degrees and 7 degrees valgus. Postoperative flexion averaged 101 degrees. There were no cases of peroneal nerve palsy or patellar dislocation. Six knees underwent revision surgery with two for sepsis, three for aseptic loosening, and one for a traumatic patella fracture. Radiographic component loosening also was seen in one knee. The probability of retention of the prosthesis was 91% (+/- 11.7%) at 13.2 years. Although the results in this group of patients seem acceptable, the rate of postoperative instability for all patients treated using this ligament balancing technique was 24%. Because of the high rate of instability, a new soft tissue release technique has been developed and is the preferred method for ligament balancing of the valgus knee during total knee arthroplasty. PMID- 9418619 TI - Continuous passive motion with accelerated flexion after total knee arthroplasty. AB - The use of continuous passive motion after total knee arthroplasty remains controversial. A new approach, starting continuous passive motion at 70 degrees to 100 degrees flexion in the recovery room (Group I) was evaluated. A randomized, prospective study of 210 consecutive total knee arthroplasties was performed at two institutions. The control population (Group II) started continuous passive motion at 0 degree to 30 degrees, and progressed toward 100 degrees flexion. Flexion at postoperative Day 3 (Group I = 82.5 degrees, Group II = 72.8 degrees), and at discharge (Group I = 89.1 degrees, Group II = 84.3 degrees) were significantly different. There was no significant difference between the groups at 4 weeks (Group I = 5.0 degrees-104.1 degrees, Group II = 5.6 degrees-102.0 degrees), 6 weeks (Group I = 2.3 degrees-104.8 degrees, Group II = 2.7 degrees-103.6 degrees), 12 weeks (Group I = 1.7 degrees-107.7 degrees, Group II = 4.7 degrees-108.2 degrees), or at 1 year (Group I = 0.5 degree-113.2 degrees, Group II = 1.8 degrees-110.5 degrees). In Group I, wound necrosis developed in one patient that required a gastrocnemius flap. This major complication was caused by a tight dressing, and not necessarily to the accelerated flexion continuous passive motion. This investigation shows that continuous passive motion using accelerated flexion allows increased flexion during the hospital stay without increased risk of complications, pain, or blood loss. This has significant implications for achieving safe, early discharge. However, no difference was found at followup of 4 weeks or greater, and this did not add significantly to the final outcome. PMID- 9418620 TI - The effect of tibial stem design on component micromotion in knee arthroplasty. AB - Rigid body mechanics with computer data acquisition and analysis techniques were used to determine the three-dimensional motions of any point on the tibial component of a total knee arthroplasty. Three stem configurations were compared: (1) no stem; (2) short stem (40 mm); and (3) long stem (75 mm). In addition, three loading conditions were analyzed for each stem configuration: (1) central loading; (2) posterior loading; and (3) medial loading. The longer stem implants were associated with increased micromotion, especially under eccentric loading. Cemented implants seemed to have more stable fixation, compared with noncemented implants. It was thought that the increased motion was secondary to a toggling of the implant under load, secondary to uneven medullary cortical contact. Overall, the results indicated that short and long stems do not enhance initial fixation with cemented or cementless implantation in routine knee arthroplasty. PMID- 9418621 TI - Tibial insert undersurface as a contributing source of polyethylene wear debris. AB - Sixty-seven ultrahigh molecular weight polyethylene tibial inserts from cementless total knee arthroplasties were retrieved at autopsy and revision surgery and analyzed for evidence of articular and nonarticular surface wear after a mean implantation time of 62.8 months (range, 4-131 months). Polyethylene cold flow and abrasive wear on the nonarticular insert surface (undersurface) were assigned a wear severity score (Grade 0-4). The severity of articular wear was assessed quantitatively and graded. Corresponding prerevision radiographs were evaluated for evidence of tibial metaphyseal osteolysis and osteolysis around tibial fixation screws. Exact nonparametric conditional inference methods were used to establish correlations between different variables and the occurrence of tibial metaphyseal osteolysis. Severe Grade 4 wear of the tibial insert undersurface was associated with tibial metaphyseal osteolysis or osteolysis around fixation screws. Time in situ statistically was related to Grade 4 undersurface wear and tibial metaphyseal osteolysis. The occurrence of tibial osteolysis was not related statistically to articular wear severity, insert thickness, or implant type. The main articulation between the femoral implant and ultrahigh molecular weight polyethylene insert has been assumed to be the primary source of polyethylene debris contributing to osteolysis and total knee arthroplasty implant failure. The undersurface of the insert is an additional source of polyethylene debris contributing to tibial metaphyseal osteolysis. To lessen polyethylene debris produced at this modular interface, the tibial implant locking mechanism should fix the insert firmly to the metal backing to decrease relative micromotion. Because motion between the insert and metal backing may be inevitable, the wear characteristics of the inner tray surface should be optimized to minimize wear debris production at this other articulation. PMID- 9418622 TI - In vivo kinematic analysis of a mobile bearing total knee prosthesis. AB - Ten normal subjects and 10 patients with a posterior cruciate retaining mobile bearing total knee replacement performed successive deep knee bends under fluoroscopy to determine tibiofemoral contact positions. At full extension the average initial contact position for the normal and mobile total knee replacement was 6.2 mm (range, 4.8 to 12 mm) anterior, and -4.4 mm (range, 3.9 to 11 mm) posterior to the sagittal tibial midplane, respectively. At 60 degrees flexion, the normal knee rolled back to -5.8 mm (range, -2.5 to -13.2 mm), whereas the mobile bearing total knee replacement rolled back to -9.2 mm (range, -4 to -17 mm). From 60 degrees to 90 degrees, normal knees rolled back to -7.8 mm (range, 5.8 to -13.8 mm), but the mobile bearing total knee replacement slid anteriorly to -5 mm (range, 2 to -12 mm). All mobile bearing total knee replacements had some form of roll back, but some slid anterior more than others. Five of 10 mobile bearing total knee replacements had some movement of the bearings while the others remained fixed. Patellar kinematics was similar to normal but reflected tibiofemoral abnormalities. PMID- 9418623 TI - Outcome of hip and knee arthroplasty in persons aged 80 years and older. AB - Recent studies have established the cost effectiveness and safety of total joint arthroplasties. As the population ages, it is important to determine whether these procedures are equally beneficial in the elderly. The short term safety and efficacy of total hip and knee arthroplasties in subjects 80 years of age and older was evaluated. Between 1988 and 1993, preoperative and postoperative physical and functional information was collected on 99 consecutive elective hip and knee arthroplasties in subjects 80 years of age or older. These data were compared with those derived from a younger otherwise matched control group. Data collected included subject demographics and characteristics, information concerning the acute and postacute hospital stay, comorbid conditions, postoperative complications, discharge disposition, Hospital for Special Surgery knee and Harris hip scores, pain scores, and functional capacity. The average age of the subjects was 83 years; osteoarthritis was the most common diagnosis; and the average followup was 25 months. Complication rates and length of stay in acute care facilities were not significantly different than for the control group. Mean preoperative Hospital for Special Surgery knee and Harris hip scores were 58 and 60, respectively, with postoperative scores of 77 and 88, respectively. Pain dramatically improved with 98% of total knee arthroplasty and 100% of total hip arthroplasty subjects reporting mild or no pain at followup. Preoperatively, none of the knee or hip subjects could walk unlimited distances. Postoperatively 51% of the total knee arthroplasty and 54% of the total hip arthroplasty subjects could walk more than five blocks; 71% of the total knee arthroplasty and 86% of the total hip arthroplasty subjects walked with a cane or no assistive device. The most dramatic postoperative functional gains were seen in the most disabled subjects. Total charges of care for patients 80 years of age and older was slightly greater than for a younger group. It was established that total joint arthroplasty can be performed safely in patients 80 years of age and older, promising excellent pain relief and improved functional outcome. PMID- 9418624 TI - Survivorship of cemented total knee arthroplasty. AB - The survivorship method of analysis was used to compare the failure rate and overall success of 2629 cemented primary total knee arthroplasties during a 22 year period by the senior surgeon. There were 215 Total Condylar prostheses with a polyethylene tibia, 265 of the Posterior Stabilized type with an all polyethylene tibia, 2036 Posterior Stabilized with a metal backed tibial component, 49 Posterior Stabilized with modular augmented components, and 64 with the Constrained Condylar system. Failure was considered revision or planned revision. The Total Condylar series had an average annual failure rate of 0.46% and a 21-year success rate of 90.77%. The Posterior Stabilized prosthesis with an all polyethylene tibia had an average annual rate of failure of 0.38% and a 16 year success rate of 94.10%, and this prosthesis with a metal backed tibial component had an annual failure rate of 0.14% and a 14-year success rate of 98.10%. The Posterior Stabilized series with modular components had an average annual rate of failure of 0.59% and a 10-year success rate of 93.63%. The Constrained Condylar knee series had an average annual failure rate of 0.26% and a 7-year success rate of 98.12%. This review represents a retrospective analysis of consecutive series of cemented, total knee arthroplasties, whose annual failure and success rates were done during differing time spans. The overall success rate was not influenced by gender, age, diagnosis, or percentage of ideal body weight. Failure was considered revision or planned revision. The best and worse case scenarios were calculated for each series. Long term results of cemented, total knee arthroplasty with a relatively conforming articular surface has been shown to be a reliable procedure with excellent survivorship. PMID- 9418625 TI - Differences in patellar tracking and knee kinematics among three different total knee designs. AB - Patellar complications are the primary reason for reoperation of the current condylar type designs. The aim of this study was to compare patellar tracking of various knee implant designs: Genesis II NexGen, and the P.F.C. Sigma Modular Knee System regarding trochlear groove center curvature. Nine unembalmed whole cadaveric lower extremities were used. The quadriceps and hamstrings were dissected into their individual muscles and loads were applied onto the muscles proximally based on the cross sectional area of the muscles. The three dimensional kinematics of the patellofemoral and tibiofemoral joint of the intact knee were measured using a 3Space tracking system. Three implants (one from each company) were implanted onto the same cadaveric knee in random order consecutively. This was done to ensure consistency of the soft tissue constraints in influencing the amount of patellar tracking. Patellar rotation, patellar tilting, patellar lateral shift and patellar displacement in relation to groove center were measured. There was no significant difference between the intact knee and the various implants regarding patellar rotation and lateral shift. However, all three prosthetic designs showed a significant degree of lateral tilting when compared with the intact knee. At 60 degrees knee flexion, the normal patella was tilted laterally to 0.44 degree +/- 2.15 degrees as compared with the Genesis II patella at 4.75 degrees +/- 4.81 degrees, the NexGen patella at 4.85 degrees +/- 4.81 degrees, and the P.F.C. Sigma patella at 4.89 degrees +/- 3.79 degrees lateral tilt. There was no difference between the intact knee compared with the resurfaced patella in patellar displacement in relation to the groove center. This study suggests the relatively similar kinematic behavior between the implant designs as compared with the intact knees. However, additional modification of implant geometry may be required to help decrease the amount of patellar tilt. PMID- 9418626 TI - Outcome implications for the timing of bilateral total knee arthroplasties. AB - Health Care Financing Administration data from 1985 to 1990 revealed 339,152 total knee arthroplasties of which 62,730 (18.6%) were bilateral procedures (simultaneous 112,922; staged 6 weeks, 4354; staged 3 months, 4524; staged 6 months, 9829; and staged 1 year 31,401). Medicare beneficiaries undergoing bilateral procedures were an average of 73 years of age; demographics revealed that among the various simultaneous and staged groups 57% to 69% were females, 90% were white, 85% to 90% had a diagnosis of osteoarthritis, and 30% to 40% were performed in rural hospitals. Between 1985 and 1990, surgical and vascular complications ranged from 2.4% to 4% and 4.1% to 6.8%, respectively, for all types of bilateral staged and simultaneous total knee arthroplasties. All differences were statistically significant. After controlling statistically for demographic variables and diagnoses, a surrogate for case mix, it was found that individuals electing simultaneous bilateral arthroplasties experienced twice the number of intensive care days than those choosing staged procedures. Days in the intensive care unit were double when done simultaneously instead of staged (0.48 versus 0.21). Nosocomial infections were similar within groups (10% versus 13%); however, wound infections were nearly half when done simultaneously (0.5% versus 1%) versus in a staged fashion. Length of stay and cost were much less for the simultaneous procedure group who were sicker as measured by the number of diagnoses. Mortality at 30 days was highest for the simultaneous procedure group (.99%) versus staged 3 or 6 months (0.30%); however, by 2 years it was close to 4% for all groups. Staging the procedure 3 to 6 months seems to offer the fewest disadvantages, is only slightly more expensive, and has the lowest mortality rate. PMID- 9418627 TI - Simultaneous bilateral versus unilateral total knee arthroplasty. Outcomes analysis. AB - One hundred consecutive, primary simultaneous bilateral total knee arthroplasties were prospectively compared with 100 consecutive, primary unilateral total knee arthroplasties in reference to relative risk, complications, cost, and need for rehabilitation. All procedures were performed using identical preoperative, intraoperative, and postoperative protocols. Postoperative confusion was approximately four times greater in the simultaneous bilateral total knee arthroplasties group (29% versus 7%), which was thought to represent an increased incidence of fat embolism. Cardiopulmonary complications were approximately three times greater after simultaneous bilateral total knee arthroplasties (14% versus 5%), and most commonly involved arrhythmias. The increased stress on the cardiopulmonary system with simultaneous bilateral total knee arthroplasties may make this procedure contraindicated in certain patients with preexisting disease. There was an approximately 17 times greater need for banked blood in the simultaneous bilateral total knee arthroplasties group (17% versus 1%), which is alarming given the persistent concerns of transfusion related disease transmission. Although the length of hospitalization was similar (6.4 days simultaneous bilateral total knee arthroplasties versus 6 days unilateral total knee arthroplasty), 89% of the patients in the simultaneous bilateral total knee arthroplasties group required a rehabilitation stay versus 45% of the patients in the unilateral total knee arthroplasty group. Total hospital charges averaged $53,168 for simultaneous bilateral total knee arthroplasties versus $32,598 for unilateral total knee arthroplasty. Total rehabilitation charges were similar. The relative cost savings implicit by doing simultaneous bilateral total knee arthroplasties seem to be at least partially offset by the approximately two times greater need for rehabilitation in this group. The true safety, efficacy, relative risk, and total cost analysis of simultaneous bilateral total knee arthroplasties demands further critical evaluation. PMID- 9418628 TI - Periprosthetic fractures of the tibia associated with total knee arthroplasty. AB - One hundred two periprosthetic tibial fractures associated with total knee arthroplasty were identified in 29 men and 73 women. Eighty-three fractures occurred postoperatively, and 19 occurred intraoperatively. Fractures were classified into four types based on location and proximity to the prosthesis. There were 61 Type I fractures, occurring at the tibial plateau; 22 Type II fractures, occurring adjacent to the prosthetic stem; 17 Type III fractures, occurring distal to the prosthetic stem; and two Type IV fractures, involving the tibial tubercle. Fracture types were additionally classified by whether the prosthesis appeared to be radiographically well fixed (A) or loose (B) at the time of fracture, or whether the fracture occurred intraoperatively (C). The majority of postoperative Types I and II fractures were Types IB and IIB, and these were treated most successfully with revision surgery. Types IIA, IIIA, and IVA fractures were managed successfully by the usual principles of tibial fracture treatment. Type IC fractures usually were managed by intraoperative fixation, Type IIC by bone grafting or external immobilization and weightbearing restrictions, and Type IIIC by conventional fracture management. This classification system provides a guide for determining the appropriate treatment for tibial fractures associated with total knee arthroplasty. PMID- 9418629 TI - Clinical pathway management of total knee arthroplasty. AB - Using a retrospective cohort study design, the authors examined complications, readmissions, morbidity and mortality, and function scores in two groups of patients attended by the same surgeon for the year before and the year after the implementation of an outcomes management program with clinical pathways for patients undergoing total knee arthroplasty at an academic health center. The effectiveness of the pathway constantly was adjusted using variance analysis and continuous quality improvement techniques. This program reduced the length of stay by 57% from a premanagement value of 10.9 +/- 5.4 days in 1994 (Group 1) to 4.7 +/- 1.4 days in 1996 (Group 2). Hospital costs (based on an inflation adjusted cost to charge ratio) for all total knees were reduced 11% from $13,328 +/- $3905 in 1994 to $11,862 +/- $4763 in 1996. Preoperative and postoperative knee scores were 41.1 +/- 16.3 and 84.2 +/- 16.0 for Group 1 and 42.5 +/- 13.0 and 87.0 +/- 10.4 for Group 2, respectively. There was no statistically significant difference between the preoperative or the postoperative knee scores of Groups 1 and 2. The application of clinical pathways, variance analysis, and continuous quality improvement toward the treatment of patients who had total knee arthroplasty at an academic health center resulted in significant savings in length of stay without adversely affecting overall outcome. PMID- 9418630 TI - Cost effectiveness and quality of life in knee arthroplasty. AB - Few studies quantitate the cost of a quality well being as produced by arthroplasty surgery. The objective was to use the Quality of Well Being Index to calculate the cost per quality of well year in knee arthroplasty surgery. The difference in Quality of Well Being Index scores before and after the intervention was calculated and multiplied by the patient's life expectancy. The procedure cost was divided by this quantity resulting in the cost of a quality well year. One hundred patients underwent a primary knee arthroplasty. There were 30 males (average age, 62 years old) and 70 females (average age, 64 years old). The calculated costs per a quality well year were $30,695 (standard deviation $90,883) at 3 months, $17,804 (standard deviation $25,888) at 6 months, $11,560 (standard deviation $11,874) at 1 year, and $6656 (standard deviation $3567) at 2 years postsurgery. Health economists consider an intervention costing less than $30,000 per quality of well year a bargain to society. Cost effectiveness of knee arthroplasty surgery compares favorably with other surgical interventions such as coronary artery bypass surgery ($5000 per quality of well year) and extremely favorable with medical treatments such as renal dialysis ($50,000.00 for the quality well year). Knee arthroplasty is a cost effective procedure and should be considered an appropriate investment by society. PMID- 9418631 TI - Opportunities for control of hospital cost for total knee arthroplasty. AB - The hospital financial records of 120 consecutive patients who underwent unilateral knee replacement surgery at one hospital during 1995 were reviewed to determine opportunities for control of hospital cost for total knee arthroplasty. The average hospital length of stay for these patients was 4.27 days (range, 3-10 days). The average hospital cost was $10,231. All 120 patients were classified under Diagnosis Related Group 209, principle procedure 81.54 primary total knee arthroplasty. Medicare paid for 70% of the patients. All payers were profitable except Medicaid and one managed care organization. When hospital cost for total knee arthroplasty was allocated to hospital service centers, 78% of the cost was attributed to the operating room, nursing units, recovery room, and pharmacy. When hospital cost for total knee arthroplasty was allocated to hospital days, 80% of the hospital cost occurred during the first 48 hours of hospitalization. Hospital reimbursement for total knee arthroplasty is primarily a prospective case price payment system. After initial cost containment efforts reduce the hospital length of stay for total knee arthroplasty to 4 to 6 days, additional control of hospital cost should focus on these areas of opportunity. PMID- 9418632 TI - Bone loss associated with the use of spacer blocks in infected total knee arthroplasty. AB - Twenty-five knees in 24 patients with infected total knee replacements were treated with debridement, component removal, and insertion of an antibiotic impregnated cement spacer block. Intravenous antibiotics were administered for 6 weeks during which time the patients' knees were kept in a knee immobilizer nonweightbearing. These cases were reviewed retrospectively to determine radiographically the amount of bone loss that occurred during the period before reimplantation. Tibial and femoral bone loss occurred frequently from invagination of the cement spacer block into the femoral and tibial cancellous bone. Tibial bone loss was present in 10 (40%) cases and averaged 6.2 mm. Femoral bone loss was present in 11 (44%) cases and averaged 12.8 mm. Bone loss was more common when spacer blocks were undersized. None of the 15 spacer blocks that were made with a small intramedullary stem displaced. Three of the remaining 10 spacer blocks made without an intramedullary stem did displace with associated bone loss. Antibiotic spacer blocks used in the two-stage treatment of infected total knee replacements can be associated with subsequent tibial and femoral bone loss. PMID- 9418633 TI - Effect of a patient management system on outcomes of total hip and knee arthroplasty. AB - Five hundred fifty-three patients undergoing hip and knee reconstructive procedures in one institution that used a patient management system were compared with a retrospective group of 340 patients undergoing similar procedures in the same institution. All procedures were performed by one surgeon and the same patient management team. Measures of length of stay, discharge disposition, and hospital charges were recorded for all patients in each subgroup of total hip arthroplasty, revision total hip arthroplasty, total knee arthroplasty, revision total knee arthroplasty, unicompartmental knee arthroplasty, and bilateral procedures. The length of stay and hospital charges were reduced significantly in all groups, whereas the percentage of patients discharged to home was unchanged. There was no significant difference in complication rates between the two groups. PMID- 9418634 TI - The need for a dual rating system in total knee arthroplasty. AB - The sequential course of the knee score and functional score of the Knee Society rating system of 276 press fit condylar modular unconstrained total knee arthroplasties performed for osteoarthritis between June 1988 and December 1992 was documented prospectively. The knee score increased significantly and stayed on a constant level from 2 years on, whereas the function score reached a maximum at 2 years and declined subsequently. Multiple regression analysis was performed testing the statistical significance and correlation of preoperative predictors with criteria at followup to determine their influence on outcome. Preoperative predictors were knee score and function score, body mass index, age, gender, patient category, and implant factors. Criteria studied were pain, knee score, and function score at 2 years followup. The function score is influenced significantly by the walking distance, age, body mass index, and patient category correlating moderately. The knee score is not affected by any of these factors. Pain was found to correlate low with the walking distance. Rating systems are influenced by numerous factors linked to the patient's general health and condition. Their impact on the overall result can be controlled by separate rating of the knee score and function score as the dual Knee Society rating system does. Scoring systems adding up knee and functional rating to an overall result should not be used. There is a need for additional improvement of total knee arthroplasty rating such as patient based evaluation and establishing reliability and validity. PMID- 9418635 TI - Irreducible isolated dislocation of the radial head. AB - Isolated dislocation of the radial head in adults is a rare injury. A 20-year-old man received a direct blow to his elbow and sustained an isolated anterior dislocation of the radial head. Open reduction was required because soft tissue interposition prevented reduction by closed manipulation. At operation the annular ligament was found to be ruptured and interposed between the articular surfaces, preventing reduction. A satisfactory result was obtained 1 year after surgery. PMID- 9418636 TI - Acute attenuation of the extensor pollicis longus tendon. A case report. AB - The current understanding of tendon biomechanics indicates that indirect injury to the tendon midsubstance requires the presence of preexisting disease during mechanical overload. This belief has been substantiated by the association of extensor pollicis longus rupture with chronic tenosynovitis caused by repetitive activity, inflammatory conditions such as rheumatoid arthritis, and minimally displaced distal radius fractures. This case of acute, traumatic, intratendinous attenuation of the extensor pollicis longus tendon offers a contradiction to the view that midsubstance tendon failure requires preexisting disease. PMID- 9418637 TI - Anticardiolipin antibodies and osteonecrosis of the femoral head. AB - The current study evaluated the prevalence of anticardiolipin antibodies, which have been associated with thrombotic phenomena, in patients with nontraumatic osteonecrosis of the hip and assessed whether the presence of such antibodies is associated with an increased risk for the development of bone necrosis. Forty consecutive patients (25 men and 15 women) with nontraumatic osteonecrosis of the hip were studied. Their ages ranged from 19 to 56 years (average, 34.3 years). Anticardiolipin antibodies were present in 37.5% (15 of 40) of the tested patients, a significantly higher rate than is seen in healthy subjects, of whom only one of 100 had low titer anticardiolipin antibodies (1%). Six of 40 patients tested positive for immunoglobulin M alone, and six of 40 patients tested positive for immunoglobulin A alone. Three of 40 patients tested positive for immunoglobulin M and immunoglobulin A isotype. The results of the current study indicate an increased incidence of anticardiolipin antibodies in patients with nontraumatic osteonecrosis of the femoral head, which may reflect that anticardiolipin antibodies play a role in the pathogenesis of bone necrosis by predisposing to thrombotic phenomena. PMID- 9418639 TI - Subtalar arthrodesis for posterior tibial tendon dysfunction and pes planus. AB - Twenty-one patients (21 feet in 18 women and three men) who were treated with subtalar arthrodesis for posterior tibial tendon dysfunction and pes planus at an average age of 60 years (range, 44-75 years) were studied. Mean duration of symptoms was 3 years. All had realignment of the calcaneus in relation to the talus, and all had screw fixation without supplemental bone graft. Average followup was 3 years (range, 2-5 years). All patients had successful union. The tibiocalcaneal angle averaged 13 degrees +/- 3.1 degrees before operation and 6 degrees +/- 1.9 degrees after operation. The lateral talometatarsal, lateral talocalcaneal, and lateral tibiotalar angles all improved significantly. Arch height (navicular height) increase averaged 5 +/- 2.7 mm. Arch length (calcaneal metatarsal) decrease averaged 4 +/- 3.2 mm. One complication occurred: delayed wound healing (excellent result). Clinical results were excellent in eight feet, good in eight, fair in four, and poor in one. Patients were satisfied with the operative result in 16 feet, satisfied with reservations in four, and dissatisfied with the operative results in one. Subtalar arthrodesis effectively corrects deformity, does not require bone graft, has a high union rate, and is associated with a low complication rate. However, 11 of the 21 patients continued to have some pain, and in patients with preexisting arthrosis of adjacent joints, symptoms may persist. PMID- 9418638 TI - Neurologic injury in the upper extremity after total hip arthroplasty. AB - The results of 7150 consecutive primary and revision total hip arthroplasties performed between 1976 and 1990 were reviewed retrospectively. Sixteen upper extremity neurologic palsies were identified in 16 patients. The incidence of upper extremity nerve palsies after total hip arthroplasty was 0.22%. There were five men and 11 women (average age, 59.5 years; range, 27-81 years). The neurologic injuries consisted of 10 ulnar palsies, four brachial plexopathies, one axillary nerve palsy, and one median nerve palsy. Patients were evaluated with respect to age, gender, preoperative diagnosis, type of procedure (primary versus revision), and surgical approach. Preoperative diagnoses included: inflammatory arthritis (11), osteoarthritis (two), avascular necrosis (one), developmental dysplasia of the hip (one), and posttraumatic arthritis (one). Fourteen of 16 patients (88%) had complete recovery. Two patients (12%) had persistent symptoms despite operative intervention. The only significant predisposing factor to developing an upper extremity neurologic injury after total hip arthroplasty was the preoperative diagnosis of an inflammatory arthropathy. Upper extremity neurologic injuries after total hip arthroplasty are rare. Patients with the preoperative diagnosis of an inflammatory arthropathy are at greater risk for experiencing upper extremity neurologic injury. The prognosis is favorable, with 88% of patients having complete recovery. Cautious induction of anesthesia and careful attention to patient positioning in the perioperative, intraoperative, and postoperative period are essential to help minimize the incidence of neurologic injuries in the upper extremity after total hip arthroplasty. PMID- 9418640 TI - Distal tibiofibular diastasis secondary to osteochondroma in a child. AB - An 18-month-old girl with a distal tibiofibular diastasis secondary to an osteochondroma was seen with a valgus deformity of the ankle. The patient underwent operative excision of the osteochondroma at the age of 2 years. At 13 year followup there was resolution of the diastasis, and the patient was free of symptoms. Early excision obviates the need for complex reconstructive surgery to correct ankle deformity later. PMID- 9418641 TI - Ankle fractures. The Lauge-Hansen classification revisited. AB - Rational treatment of ankle fractures requires knowledge of the extent of bone and soft tissue injury. Although the Lauge-Hansen classification attempts to do this by relating specific fracture patterns to injury mechanism, the experimental underpinning for this classification has not been reexamined rigorously using modern experimental methods. This study examines the hypothesis that the clinically occurring supination and external rotation injury pattern does not result from the mechanism described by Lauge-Hansen. Thirty-two anatomic specimen ankles were mounted on an MTS machine for combined axial loading with external rotation to failure testing. A foot plate supinated the foot 25 degrees. Testing was performed with the ankle at neutral, 25 degrees plantar flexed, 10 degrees to 15 degrees dorsiflexed, and in 6 degrees to 8 degrees leg valgus. Pure supination and external rotation with the ankle in neutral did not result in the Lauge Hansen supination and external rotation type fractures. This outcome was not altered if the ankle specimens initially were placed in plantar flexion or dorsiflexion. The addition of a valgus load, which pushes the talus laterally against the fibula, resulted in the classic Lauge-Hansen supination and external rotation type fracture. All specimens had an isolated lateral injury or a lateral injury that preceded medial injury. PMID- 9418642 TI - Prognostic factors among 130 patients with osteosarcoma. AB - One hundred thirty patients with high grade osteosarcoma were enrolled in a randomized prospective multidisciplinary treatment that included intraarterial chemotherapy, local irradiation, limb salvage surgery, and prophylactic whole lung irradiation. The patients were evaluated to stage the prognostic factors. In a multivariate analysis, a minimal level of serum lactic acid dehydrogenase less than 300 IU/L showed a significant prognostic value. The history of trauma before diagnosis of disease, local irradiation of the affected site, histologic response to preoperative multidisciplinary therapy, and prophylactic whole lung irradiation were associated with significantly better prognosis in the log rank test. Patient age, site of the primary tumor, presentation of fracture, pathologic subtype, signs and symptoms, serum alkaline phosphatase level, and erythrocyte sedimentation rate were not found to be prognostic factors. The 9 year survival rate of the whole group was 55%. PMID- 9418643 TI - Anatomy of the posterior interosseous nerve in relation to fixation of the radial head. AB - The relationship of the posterior interosseous nerve to the radial neck as it relates to internal fixation of radial head fractures was studied in 50 fresh anatomic specimen arms. After a standard posterolateral approach, blind subperiosteal dissection was performed distally until a 4-cm minifragment plate could be placed on the shaft of the radius. Dissection of the radial nerve was performed under loupe magnification. In only one (2%) arm did the posterior interosseous nerve lie directly on the radius. The average distance from the posterior interosseous nerve to the plate was 5 +/- 1.2 mm. The nerve was not damaged by placement of the plate in any case. In two (4%) cases the nerve play on top of the distal portion of the plate within the supinator with the forearm in neutral rotation. The average distance from the radial head to the origin of the posterior interosseous nerve was 1.2 +/- 1.9 mm, with the takeoff being proximal to the radial head in 31 cases. The muscular branch to the extensor carpi radialis longus was located 7.1 +/- 1.8 mm from the radial head. These findings suggest that pronation of the forearm with blind subperiosteal dissection for plate placement does not place the posterior interosseous nerve at significant risk for structural injury. However, as with any approach done in the region of the nerve, caution should be used to avoid tension on the nerve that could lead to physiologic injury. PMID- 9418644 TI - Use of bone morphogenetic protein 2 on ectopic porous coated implants in the rat. AB - The ability of recombinant human bone morphogenetic protein 2 to remain osteoinductive and stimulate appositional bone formation on a porous coated implant was tested in a rat quadriceps muscle pouch. Implants with or without hydroxyapatite were used to compare the effects on bone formation of two different does (23 micrograms or 46 micrograms) of recombinant human bone morphogenetic protein 2 against controls as evidenced by contact radiography, histologic examination, and backscatter scanning electron microscopic analysis. Cylindrical plasma sprayed porous titanium implants were placed bilaterally within a muscle pouch surgically created in 48 Lewis rats. Implants treated with recombinant human bone morphogenetic protein 2 formed significantly more bone than did control implants independent of the dose or presence of hydroxyapatite. In all implants with bone formation, osteoinduction via endochondral ossification began within 7 days. By 21 days, cartilage largely was replaced by bone and marrow. The results of this ectopic, nonweightbearing in vivo assay suggest that recombinant human bone morphogenetic protein 2 remains biologically active after application to a titanium implant and may be used to enhance appositional bone formation by direct application to the implant surface. PMID- 9418645 TI - Neuropeptides in heterotopic bone induced by bone matrix in immunosuppressed rats. AB - The effects of cyclosporin A on the occurrence of neuroendocrine peptides in bone induced by demineralized allogeneic and xenogeneic bone matrix were studied in rats. Cyclosporin A enhanced bone induction in demineralized allogeneic bone matrix implants by 40% to 50% at 4 weeks, whereas there was no difference to the control group at 8 weeks. In demineralized xenogeneic bone matrix implants there was virtually no cartilage or bone formation at 4 weeks, but some bone and cartilage formation was seen at 8 weeks. In both cyclosporin A treated groups the net bone formation in demineralized xenogeneic bone matrix implants was increased four to five times at 4 weeks. Cyclosporin A treatment did not alter the temporal occurrence or distribution of neuropeptide containing nerve fibers in the bone induced by allogeneic bone matrix. Fibers containing substance P, calcitonin gene related peptide, neuropeptide Y, vasoactive intestinal peptide, and tyrosine hydroxylase were detected in the ossicles of cyclosporin A treated and control rats. In the xenogeneic bone matrix of the control group, no immunoreactive nerve fibers could be detected at 4 weeks, but at 8 weeks all five neuropeptides were detected. However, after cyclosporin A treatment immunoreactive nerve fibers could be seen at 4 weeks in the demineralized xenogeneic bone matrix implants. Thus, immunologic properties of the inductive matrix affect the yield of mineralized bone and the degree of innervation. Cyclosporin A decreases the immune response and enhances the formation of bone and the number of transmitter identified nerves in demineralized xenogeneic bone matrix induced ossicles. PMID- 9418646 TI - The Marshall R. Urist Young Investigator Award. Autogenous flexor tendon grafts. Biologic mechanisms for incorporation. AB - To examine the hypothesis that different types of dense regular connective tissue may have different repair mechanisms within the synovial space, intrasynovial and extrasynovial autogenous donor flexor tendon grafts were placed within the synovial sheaths of the medial and lateral forepaw digits of dogs. Histologic, ultrastructural, biochemical, and biomechanical analyses were done between 10 days and 6 weeks after tendon grafting. Intrasynovial tendon grafts remained viable when transferred to the synovial space and appeared to heal through an intrinsic process with preservation of the gliding surface and improved functional characteristics. Extrasynovial tendon grafts functioned as a scaffolding for the early ingrowth of new vessels and cells. Early cellular necrosis consistently was followed by the ingrowth of fibrovascular adhesions from the periphery. The formation of dense peripheral adhesions, obliterating the gliding surface of the tendon, led to diminished tendon excursion and proximal interphalangeal joint rotation. PMID- 9418647 TI - Refusal to walk in a 3-year-old girl. PMID- 9418648 TI - It's time for action. PMID- 9418649 TI - The DASH to lower blood pressure. PMID- 9418650 TI - Disheartening developments on the world stage. PMID- 9418651 TI - The accidental patient. PMID- 9418652 TI - A practical foundation. PMID- 9418653 TI - Mendel might get dizzy. PMID- 9418654 TI - Less hype, more hope. PMID- 9418655 TI - The heart of the matter. PMID- 9418656 TI - First the bad news.... PMID- 9418657 TI - Breathing easier. PMID- 9418658 TI - At the cutting edge. PMID- 9418659 TI - Progress, frustration and controversy. PMID- 9418660 TI - Progress in prostate cancer. PMID- 9418661 TI - Death: a rewarding experience? PMID- 9418662 TI - Human values in health care: trying to get it right. PMID- 9418663 TI - Consent and discontent. PMID- 9418664 TI - All in the family. PMID- 9418665 TI - One step forward, two steps back. PMID- 9418666 TI - The latest word on emerging pathogens. PMID- 9418667 TI - The microbes strike back. PMID- 9418668 TI - Immunization. Point, counterpoint. PMID- 9418669 TI - Women's health comes of age. PMID- 9418670 TI - Cracking the glass ceiling. PMID- 9418671 TI - What's up in medical informatics? PMID- 9418672 TI - This business called medicine. PMID- 9418673 TI - Wrong answers at the wrong time? PMID- 9418674 TI - You can go home again. PMID- 9418675 TI - Speaking from the heart. PMID- 9418676 TI - Where's the evidence? PMID- 9418677 TI - Factors in low birth weight. PMID- 9418678 TI - Licence plates for drugs. PMID- 9418679 TI - Where does our duty lie? PMID- 9418680 TI - Is it ethical to forgo treatment? PMID- 9418681 TI - Mifepristone (RU486): a review. AB - OBJECTIVE: To review the literature concerning the mechanism of action and pharmacodynamics of mifepristone (RU486), potential new uses of RU486, and its current use not only as an abortifacient but also as therapy for endometriosis, leiomyoma, breast cancer, and meningioma. DATA IDENTIFICATION AND SELECTION: Studies that relate to RU486 were identified through a MEDLINE search. CONCLUSION(S): RU486 is an 11 beta-dimethyl-amino-phenyl derivative of norethindrone with a high affinity for P and glucocorticoid receptors. The receptor binding is not followed by transcription of P-dependent genes. Mifepristone effectively blocks P receptors in the placenta, resulting in the termination of pregnancy. In addition, it has been used in the treatment of leiomyomata, endometriosis, advanced breast cancer, and meningioma. It is a powerful tool to study the molecular action of P and in the future may be used as an estrogen-free contraceptive. PMID- 9418682 TI - Future trends in infertility treatment: challenges ahead. PMID- 9418683 TI - Does dong quai have estrogenic effects in postmenopausal women? A double-blind, placebo-controlled trial. AB - OBJECTIVE: To evaluate possible estrogenic effects of dong quai on vaginal cells and on endometrial thickness in postmenopausal women. DESIGN: Double-blind, randomized, placebo-controlled clinical trial. SETTING: Department of Obstetrics and Gynecology in a large health maintenance organization (HMO). PATIENT(S): Seventy-one postmenopausal women (mean age [+/- SD], 52.4 +/- 6 years) who had follicle-stimulating hormone levels (third-generation assay) of > 30 mIU/mL with hot flashes. INTERVENTION(S): Subjects were randomized to treatment with either dong quai or placebo for 24 weeks. MAIN OUTCOME MEASURE(S): Endometrial thickness was measured by transvaginal ultrasonography; vaginal cells were evaluated for cellular maturation; menopausal symptoms were evaluated by reviewing the Kupperman index and the diary of vasomotor flushes. RESULT(S): We observed no statistically significant differences between groups in endometrial thickness, in vaginal maturation index, in number of vasomotor flushes, or in the Kupperman index. CONCLUSION(S): Used alone, dong quai does not produce estrogen-like responses in endometrial thickness or in vaginal maturation and was no more helpful than placebo in relieving menopausal symptoms. PMID- 9418684 TI - Effects of oral glucose administration on plasma growth hormone levels in women with polycystic ovary syndrome. AB - OBJECTIVE: To evaluate the sensitivity of GH secretion to the suppressive effect of oral glucose administration in women with polycystic ovary syndrome (PCOS). DESIGN: Comparison of the GH response to an oral glucose load in women with PCOS and in weight-matched normally menstruating women (controls). SETTING: Reproductive endocrinology unit. PATIENT(S): Eighteen obese and 11 nonobese patients and 10 obese and 10 nonobese controls. INTERVENTION(S): After an overnight fast, each woman underwent a 75-g, 3-hour oral glucose tolerance test (OGTT). MEAN OUTCOME MEASURE(S): Growth hormone, glucose, and insulin responses to OGTT. RESULT(S): No significant differences in the glycemic and insulinemic responses were found between the patients and the weight-matched controls. No decrease in plasma GH was observed in both obese and nonobese patients and in obese controls during the OGTT, whereas a significant GH decrease occurred in nonobese controls 60 and 120 minutes after glucose intake. CONCLUSION(S): Oral glucose administration was unable to suppress GH levels in nonobese as well as in obese women with PCOS and in obese control women. These data suggest that both PCOS and obesity are associated with a reduced sensitivity of GH secretion to glucose suppression. PMID- 9418685 TI - Altered binding of serum thyroid hormone to thyroxine-binding globulin in women with functional hypothalamic amenorrhea. AB - OBJECTIVE: To further characterize hypothyroidemia with decreased serum concentrations of total triiodothyronine and total T4 in women with functional hypothalamic amenorrhea. DESIGN: Cohort study. SETTING: University of California San Diego Clinical Research Center. PATIENT(S): 8 women with functional hypothalamic amenorrhea (hypogonadotropic or normogonadotropic amenorrhea of at least 6 months duration) and 9 normal cycling women in the early follicular phase of their cycles. INTERVENTION(S): 24-hour frequent blood sampling. MAIN OUTCOME MEASURE(S): Comparison of levels of thyroid hormones and binding proteins between functional hypothalamic amenorrhea and normal cycling women. Measurements of serum free T4, free triiodothyronine, total T4, total triiodothyronine, reverse triiodothyronine, thyroid-binding globulin, albumin, and prealbumin levels and determination of T4 binding and binding affinity to each of the three binding proteins. RESULT(S): The results confirmed reduced levels of total triiodothyronine and total T4, but revealed no significant difference in free triiodothyronine and free T4, as well as reverse triiodothyronine, levels between functional hypothalamic amenorrhea and normal cycling women. Although serum levels of thyroid hormone-binding proteins were similar between normal cycling women and functional hypothalamic amenorrhea, a significant decrease in T4 bound to thyroid-binding globulin along with a decrease in apparent affinity of thyroid binding globulin for T4 was present in functional hypothalamic amenorrhea. No differences in prealbumin- or albumin-T4 interactions were found. CONCLUSION(S): In functional hypothalamic amenorrhea, a reduced thyroid-binding globulin binding affinity appears to explain the disparity between normal levels of free triiodothyronine, free T4, and binding proteins in the face of reduced levels of total triiodothyronine and total T4. PMID- 9418686 TI - The effects of clomiphene citrate on normally ovulatory women. AB - OBJECTIVE: To investigate the efficacy of clomiphene citrate (CC) on normally ovulatory women who complained of infertility. DESIGN: A randomized study. SETTING: University Hospital. PATIENT(S): Thirty-three normally ovulatory women with unexplained infertility. INTERVENTION(S): Eighteen women received CC at a 50 mg dosage. Fifteen women received no ovulation-induction drugs. MAIN OUTCOME MEASURES: The pregnancy rate (PR) per patient, the PR per cycle, and the cumulative pregnancy rate. RESULT(S): Seven patients in the CC group stopped taking CC, and observations were terminated because of antiestrogenic effects. The pregnancy rate (PR) per patient and the PR per cycle were significantly decreased (P < 0.005) in the CC group (4 of 18 [22.2%] and 4 of 66 [6.1%], respectively) than in the spontaneous group (11 of 15 [73.3%] and 11 of 52 [21.2%], respectively). Kaplan-Meier tests showed that the cumulative pregnancy rate in the CC group was significantly lower than in the spontaneous group (P < 0.05). Five of seven patients who had stopped taking CC became pregnant in spontaneous cycles. CONCLUSION(S): Administration of CC to normally ovulatory women is not efficacious in terms of increasing the pregnancy rate. PMID- 9418687 TI - Comparison of finasteride versus spironolactone in the treatment of idiopathic hirsutism. AB - OBJECTIVE: To compare the efficacy of finasteride and spironolactone in the treatment of idiopathic hirsutism. DESIGN: Prospective, randomized, single-blind study. SETTING: A tertiary hirsutism clinic. PATIENT(S): Forty women with idiopathic hirsutism were selected. INTERVENTION(S): Patients were assigned randomly to receive either 5 mg of finasteride or 100 mg of spironolactone for 9 months. MAIN OUTCOME MEASURE(S): Hirsutism scores were measured according to the Ferriman-Gallwey scoring system, and side effects were monitored for 9 months of treatment. Blood samples were taken at each visit for assessment of endocrine, biochemical, and hematologic parameters. RESULT(S): Hirsutism scores were decreased significantly in both groups at the end of 9 months. The mean percent change (+/- SD) in hirsutism scores in the finasteride and spironolactone groups was as follows: 5.91% +/- 7.18% and 20.60% +/- 12.59% at 3 months, 10.61% +/- 12.18% and 32.57% +/- 15.68% at 6 months, and 15.15% +/- 15.38% and 42.36% +/- 12.31% at 9 months, respectively. There was a significantly better response with spironolactone treatment at the end of 9 months. Eleven (55%) of 20 patients in the spironolactone group experienced side effects. However, none of them stopped treatment because of side effects. CONCLUSION(S): The present data suggest that both finasteride and spironolactone are effective in the treatment of idiopathic hirsutism. However, it appears that the spironolactone group responded significantly better. PMID- 9418688 TI - Delay of gonadotropin stimulation in patients receiving gonadotropin-releasing hormone agonist (GnRH-a) therapy permits increased clinic efficiency and may enhance in vitro fertilization (IVF) pregnancy rates. AB - OBJECTIVE: To promote an even temporal distribution of patients starting IVF cycles at our center, patients undergoing GnRH agonist (GnRH-a) suppression frequently delay the start of gonadotropin stimulation. Our objective was to analyze the effect that the delay of initiation of gonadotropin stimulation has on outcome parameters in this population. DESIGN: Retrospective analysis. SETTING: A tertiary referral reproductive medicine unit. PATIENT(S): Patients undergoing IVF cycles on long GnRH-a protocols. INTERVENTION(S): Patients were treated with either a "standard-dose" or "low-dose" leuprolide acetate protocol initiated in the mid-luteal phase. MAIN OUTCOME MEASURE(S): Delay time, clinical pregnancy rate, ongoing pregnancy rate, cancellation rate. RESULT(S): Analysis of the overall group revealed associations between stimulation delay and decreases in stimulation duration and the number of gonadotropin ampules administered. Weighted linear regression analyzes revealed statistically positive relationships between delay time and both clinical pregnancy rates and ongoing pregnancy rates, despite a positive relationship between delay time and cancellation rates. Analysis of the standard-dose and low-dose subgroups revealed that the enhancement of pregnancy rates was attributable primarily to patients in the standard-dose protocol. CONCLUSION(S): Delay of gonadotropin stimulation while patients are receiving GnRH-a therapy allows for increased clinic efficiency. There appears to be an enhancement of clinical and ongoing pregnancy rates for the standard-dose leuprolide acetate protocol that is associated with stimulation delay. PMID- 9418689 TI - Exogenous gonadotropin stimulation is associated with increases in serum androgen levels in in-vitro fertilization-embryo transfer cycles. AB - OBJECTIVE: To examine serum androgen profiles in women undergoing ovarian stimulation with exogenous gonadotropins in the setting of IVF-ET. DESIGN: Prospective study. SETTING: University hospital IVF-ET program. PATIENT(S): Seventeen ovulatory women undergoing IVF-ET for endometriosis, male factor infertility, or tubal disease. INTERVENTION(S): A standard long protocol of GnRH agonist (GnRH-a) pretreatment (1 mg of leuprolide acetate SC for 10 days) was administered before ovulation induction with a urinary gonadotropin preparation. MAIN OUTCOME MEASURE(S): After 10 days of GnRH-a treatment and on the day of hCG administration, serum concentrations of LH, T, androstenedione (A), sex hormone binding globulin (SHBG), and DHEAS; the free androgen index (T/SHBG); and the number of follicles, oocytes, and embryos were assessed. RESULT(S): Serum samples after 10 days of GnRH-a treatment showed incomplete LH suppression. While continuing the agonist during ovarian stimulation, LH values were suppressed further. However, serum T and A concentrations and the free androgen index showed a significant increase (samples drawn just before hCG administration). Serum T levels after 10 days of GnRH-a (before the administration of exogenous gonadotropins) were correlated negatively with the subsequent number of embryos. CONCLUSION(S): Serum LH suppression with a conventional regimen of GnRH-a is incomplete in this heterogeneous group of ovulatory women. Exogenous gonadotropin stimulation results in a marked increase in ovarian androgen secretion. PMID- 9418690 TI - Oocyte quality in patients with severe ovarian hyperstimulation syndrome. AB - OBJECTIVE: To study the oocyte quality in patients with ovarian hyperstimulation syndrome (OHSS). DESIGN: Retrospective study. SETTING: The Egyptian IVF-ET Center. PATIENT(S): Forty-two patients who developed severe OHSS (group A) were studied for the mean number of oocytes retrieved, percentage of high-quality oocytes, embryo quality, and fertilization, implantation, and pregnancy rates; these patients were compared with an age-matched control group who did not develop OHSS (group B; n = 183) after superstimulation for IVF or intracytoplasmic sperm injection. INTERVENTION(S): In vitro fertilization and ICSI. MAIN OUTCOME MEASURE(S): Fertilization and pregnancy rates. RESULT(S): In group A, the mean number of oocytes retrieved was significantly higher, whereas the percentage of high-quality oocytes and the fertilization rate were significantly lower than that in group B. There were no statistically significant differences in the quality of embryos transferred or the implantation or pregnancy rate between the groups. The percentage of high-quality oocytes and the fertilization rate were significantly lower in patients with polycystic ovaries (PCO) in both groups. CONCLUSION(S): The inferior quality and maturity of oocytes in OHSS reduced the fertilization rate but did not affect the quality or the number of embryos transferred or the pregnancy rate. The effect on oocyte quality could be due to the prevalence of PCO in this group of patients. PMID- 9418691 TI - Does hydrosalpinx reduce the pregnancy rate after in vitro fertilization? AB - OBJECTIVE: To determine the effect of hydrosalpinges on the pregnancy rate in an IVF program. DESIGN: Multicentric retrospective analysis of clinical and laboratory data. SETTING: Two assisted reproductive technology centers in university hospitals. PATIENT(S): Four hundred forty-three women, under 38 years of age, with pure tubal infertility. The patients were classified into the following five groups: bilateral hydrosalpinges (n = 37), unilateral hydrosalpinx (n = 54), bilateral tubal occlusion (n = 207), unilateral tubal occlusion (n = 55), and severe tubal disease without complete occlusion (n = 90). MAIN OUTCOME MEASURE(S): Pregnancy and implantation rates. RESULT(S): The pregnancy and the implantation rates per transfer (12.3% and 5.4%) obtained by women with bilateral hydrosalpinges are significantly lower than the rates (means = 23.1% and 12%) for all other tubal infertility groups. CONCLUSION(S): Bilateral hydrosalpinges have a deleterious effect on the outcome of IVF program. PMID- 9418692 TI - Diagnosis of ectopic pregnancy after in vitro fertilization and embryo transfer. AB - OBJECTIVE: The combination of transvaginal sonography and serum hCG measurement is reliable in the diagnosis of ectopic pregnancy (EP) in spontaneous pregnancies. In patients who became pregnant through IVF-ET, transfer of multiple embryos after IVF could be responsible for the different performance of these tests. We evaluated the discriminative capacity of transvaginal sonography in combination with hCG measurement in the diagnosis of EP after IVF-ET. DESIGN: Prospective cohort study. SETTING AND PATIENT(S): Consecutive patients, pregnant through IVF-ET, who presented with clinically suspected EP. INTERVENTION(S): Transvaginal sonography, serum hCG measurement at 6, 9, and 15 days after ET and after a negative transvaginal sonography. MAIN OUTCOME MEASURE(S): Ectopic pregnancy confirmed at laparoscopy. RESULT(S): Between September 1993 and May 1996, 86 women were included in the study, of whom 24 had an EP. Transvaginal sonography identified 46 intrauterine pregnancies and 5 EPs, but serum hCG could not diagnose EPs in patients in whom transvaginal sonography did not show a gestational sac. Serum hCG measurement 9 days after ET could identify pregnancy failure with 100% specificity at a cut-off value of 18 IU/L, but it could not identify patients with EP with enough certainty to justify immediate treatment. CONCLUSION(S): We recommend single serum hCG measurement 9 days after ET to discriminate between viable and nonviable pregnancies. Transvaginal sonography can be postponed until 5 weeks after ET, except for patients with abdominal pain and/or vaginal bleeding, or patients with a serum hCG level of < 18 IU/L. PMID- 9418693 TI - First and subsequent pregnancies after tubal microsurgery: evaluation of the fertility index. AB - OBJECTIVE: To determine the number of children born after a tubal microsurgical operation and to evaluate the fertility index, a long-term measure of reproductive potential. DESIGN: A case series involving a follow-up questionnaire. SETTING: A tertiary care university hospital. PATIENT(S): Three hundred twelve women undergoing microsurgery for tubal disease. INTERVENTIONS: A range of open microsurgical procedures including reversal of sterilizations. MAIN OUTCOME MEASURE(S): Cumulative pregnancy rates to the first and second normal pregnancies and calculation of the fertility index. RESULT(S): The 2-year cumulative pregnancy rates (probability +/- SE) for a first normal pregnancy for proximal disease, distal disease, tubal reanastomoses, and tubal adhesions were 0.51 +/- 0.05, 0.29 +/- 0.06, 0.47 +/- 0.06, and 0.30 +/- 0.07, respectively. Of the 288 (92%) women responding to the questionnaire, 142 women had at least one child. Of the 100 women who wanted a second child, 68 succeeded, the proportions being similar in each surgery category. The fertility index described restoration of normal fertility in 30%, 12%, 34%, and 23% for women with proximal disease, distal disease, anastomotic procedures, and adhesion disease, respectively. CONCLUSION(S): The fertility index is a useful measure of long-term reproductive potential. The high recurrent pregnancy rate emphasizes the value of microsurgery in restoring normal fertility to some women. PMID- 9418694 TI - Tubal patency and fertility outcome after expectant management of ectopic pregnancy. AB - OBJECTIVE: To study tubal patency and fertility outcome of patients with expectantly managed ectopic pregnancy (EP). DESIGN: A prospective study. SETTING: Department of Obstetrics and Gynecology, Turku University Central Hospital, Turku, Finland. PATIENT(S): Thirty patients who wanted to become pregnant again were treated successfully by expectant management of EP. INTERVENTION(S): Patients were examined with posttreatment hysterosalpingography, and they filled out a questionnaire about their subsequent pregnancies. MAIN OUTCOME MEASURE(S): Free passage through open lumen showing tubal patency; number of full-term pregnancies and EPs revealing relative rate of subsequent fertility. RESULT(S): A free passage through the diseased tube was seen in 93% of the cases (28 of 30). Two of the 24 patients (8.4%) who wanted to become pregnant had an obstruction in the diseased tube. One (4.2%) of them had a normal opposite tube and later had a normal pregnancy. The other (4.2%) had an obstruction in both tubes and subsequently had a repeat EP. One of the 6 patients had an EP (this patient did not want to become pregnant and did not use contraception). However, her posttreatment hysterosalpingography was normal. In total, the subsequent pregnancy rate was 88% (21 of 24), and the rate of repeat EP was 4.2% (1 of 24). CONCLUSION(S): Patients who are treated with expectant management have a good long-term fertility outcome. Spontaneous regression of EP does not lead to increased harm or damage to the tube, i.e., the risk for repeat EP is low. PMID- 9418695 TI - Simplified therapy for Asherman's syndrome. AB - OBJECTIVE: To evaluate a technique that converts a blind hysteroscopic procedure to a "septum" division. DESIGN: Open noncomparative clinical study. SETTING: Tertiary care center. PATIENT(S): Six women with Asherman's syndrome; five with complete and one with incomplete obliteration of the uterine cavity. INTERVENTION(S): The patients underwent recreation of the uterine cavity by the hysteroscopic-laparoscopic technique described to establish the correct dissection plane. MAIN OUTCOME MEASURE(S): The ability to reestablish the uterine cavity; postoperative resumption of menses and fertility. RESULT(S): In all patients, the cavity of the uterus was restored; menses resumed in all women who were previously amenorrheic; and 5 women conceived, of whom four had live births and one a missed abortion. At hysteroscopy, two women incurred perforations and in another hemorrhage occurred. CONCLUSION(S): This technique appears to be effective and safe for the reconstruction of a functional endometrial cavity in women with Asherman's syndrome. PMID- 9418696 TI - The retractile testis can be a cause of adult infertility. AB - OBJECTIVE: To determine the functional state of the testes of young adults treated for small and hypotrophic retractile testis at prepubertal age by orchiopexy and/or hormonal therapy and the functional state of the testes of adults with retractile testis. DESIGN: Spermiogram and transmission electron microscopy (TEM) study of the semen. Light microscopic and TEM studies of semen and testicular biopsies of adult with retractile testis were performed. SETTING: Division of Pediatric Surgery in an academic environment. PATIENT(S): Thirty eight young adults (mean age, 18 years) treated for retractile testis at prepubertal age and seven adults (mean age, 28 years) with retractile testis. INTERVENTION(S): Two cycles of hCG, followed by surgical therapy (orchiopexy) when unsuccessful. MAIN OUTCOME MEASURE(S): Fertility of young adults treated for retractile testis at prepubertal age. RESULT(S): Only 8 of 38 (21%) young adults had normal spermiograms. Five of 38 (13%) were azoospermic and 25 of 38 (66%) were oligoasthenozoospermic with ultrastructural signs of altered maturation of the sperm and a higher number of atypical forms. Of the adults with retractile testis, 2 of 7 (28.5%) were normal, 3 of 7 (43%) were oligoasthenozoospermic, and 2 of 7 (28%) were azoospermic. CONCLUSION(S): Our data support the hypothesis that prepubertal retractile testis showing signs of reduced consistency and size is a risk factor for adult infertility and requires treatment. PMID- 9418697 TI - No evidence for decreasing semen quality in four birth cohorts of 1,055 Danish men born between 1950 and 1970. AB - OBJECTIVE: To compare the quality of semen in 1,055 Danish men born between 1950 and 1970 who are assumed to represent a random sample of the Danish male population of fertile age. DESIGN: Retrospective review of data on semen quality at the time of the female partner's first IVF treatment. SETTING: The Fertility Clinic, Odense University Hospital, Odense, Denmark. PATIENT(S): One thousand fifty-five male partners of women with tubal infertility who were referred for IVF treatment consecutively during the period 1990-1996. INTERVENTION(S): Analysis of the semen samples delivered and used in connection with the couples' first IVF treatment. MAIN OUTCOME MEASURE(S): Year of birth, age at time of sample collection, sperm concentration, and semen volume. RESULT(S): The mean sperm concentration (+/- SD) was 183.7 x 10(6) mL and the mean semen volume (+/- SD) was 3.9 mL. A considerable variation in both parameters was found from year to year, but no significant change occurred in either parameter throughout the entire period. When four birth cohorts were compared, a later year of birth was not associated with any change in sperm concentration or semen volume. CONCLUSION(S): Sperm concentration and semen volume were not related to year of birth, indicating that sperm quality has not changed in the Danish male population during the last 20-30 years. PMID- 9418698 TI - In situ end-labeling of human testicular tissue demonstrates increased apoptosis in conditions of abnormal spermatogenesis. AB - OBJECTIVE: To determine, using an in situ end-labeling technique, whether the frequency of apoptosis is increased in testis biopsy specimens that demonstrate abnormal spermatogenesis. DESIGN: Immunohistochemical analysis was performed on archived paraffin-embedded testis biopsy specimens. Apoptotic indices, defined as the number of apoptotic bodies per the total number of cells or the number of Sertoli cells, were calculated after counting all the intratubular spermatogenic cells and Sertoli cells in 20 tubules. SETTING: Major academic male factor infertility clinic. PATIENT(S): Forty-eight testis biopsy specimens were obtained for routine clinical purposes from 38 men with azoospermia or severe oligozoospermia. INTERVENTION(S): In situ end-labeling was performed on archived paraffin-embedded testis biopsy specimens using terminal deoxynucleotidyl transferase. MAIN OUTCOME MEASURE(S): Apoptotic indices. RESULT(S): Significantly increased apoptotic indices were observed in patients with spermatocyte arrest, spermatid arrest, and hypospermatogenesis compared with patients with normal spermatogenesis and the Sertoli cell-only pattern. CONCLUSION(S): In situ end labeling of testis biopsy specimens from infertile men demonstrates increased apoptosis in maturation arrest and hypospermatogenesis states compared with normal spermatogenesis and the Sertoli cell-only pattern. This unique observation implicates a prominent role for this form of programmed cell death in the pathophysiology of maturation arrest and hypospermatogenesis states. PMID- 9418699 TI - Follow-up report on a randomized controlled trial of laser laparoscopy in the treatment of pelvic pain associated with minimal to moderate endometriosis. AB - OBJECTIVE: To assess the longer term efficacy of laparoscopic laser surgery in the treatment of painful pelvic endometriosis and to observe the natural history of the disease at second-look laparoscopy in a control group. DESIGN: One-year follow-up of a prospective, randomized, double-blind controlled trial. SETTING: A referral center for the laparoscopic laser treatment of endometriosis. PATIENT(S): Sixty-three patients with pelvic pain and minimal to moderate endometriosis. INTERVENTION(S): After the 6-month follow-up visit, the randomization code was broken, and follow-up was continued to 1 year. Symptomatic patients were offered second-look laser laparoscopy. MAIN OUTCOME MEASURE(S): Continued symptom relief at 1 year after treatment and findings at second-look laparoscopy in symptomatic controls. RESULT(S): Symptom relief continued at 1 year in 90% of those who initially responded. All symptomatic controls had a second-look procedure, with 7 (29%) showing disease progression, 7 (29%) showing disease regression, and 10 (42%) having static disease. CONCLUSION(S): The benefits of laser laparoscopy for painful pelvic endometriosis are continued in the majority of patients at 1 year. Untreated painful endometriosis will progress or remain static in the majority of patients but will spontaneously improve in others. PMID- 9418700 TI - "Crushed glass" appearance of particles observed in roentgenograms after hysterosalpingography as an indicator of pelvic abnormalities. AB - OBJECTIVE: To evaluate the diagnostic value of the crushed glass appearance of particles observed on roentgenogram after hysterosalpingography in patients with pelvic abnormalities. DESIGN: Retrospective study of 32 patients whose roentgenogram had particles with a crushed glass appearance and 16 patients without crushed glass appearance, with laparoscopy conducted to determine the location and severity of the pelvic disease. SETTING: Fujita Health University Hospital, Aichi, Japan. PATIENT(S): A total of 240 patients underwent hysterosalpingography to determine the cause of infertility. MAIN OUTCOME MEASURE(S): We compared the location of the pelvic endometriosis and/or inflammation that was observed on roentgenogram as the crushed glass appearance versus definitive findings at laparoscopy. RESULT(S): A total of 30 of the 32 cases (93.8%) with the crushed glass appearance were confirmed as having pelvic abnormalities, such as endometriosis (73.3%) and pelvic inflammation (26.7%). The rate of the concurrence of the location of the crushed glass appearance observed on the film and that of the lesions verified by laparoscopy was 66.7%. Histopathological examination showed that specimens taken from endometriosis or inflammatory lesions that comprised the crushed glass appearance were lacking in epithelium in proportion to the size of the particles. CONCLUSION(S): Identification of the crushed glass appearance of particles was a useful noninvasive method of detecting pelvic abnormalities, such as pelvic endometriosis and inflammation, in infertile women. PMID- 9418701 TI - Mutation analysis of the gonadotropin-releasing hormone receptor gene in idiopathic hypogonadotropic hypogonadism. AB - OBJECTIVE: To determine if GnRH receptor mutations occur in patients with idiopathic hypogonadotropic hypogonadism. DESIGN: Patients and controls were studied by molecular genetic analysis. SETTING: A tertiary medical center setting. PATIENT(S): Twenty-four patients with idiopathic hypogonadotropic hypogonadism and 20 controls. INTERVENTION(S): Deoxyribonucleic acid from all individuals was analyzed by Southern blot analysis and denaturing gradient gel electrophoresis. Genomic DNA was digested with restriction enzymes, and Southern blots and denaturing gradient gel blots were constructed. Blots were hybridized with the GnRH receptor complementary DNA probe. The DNA sequencing was performed on samples from two representative patients. MAIN OUTCOME MEASURE(S): Gonadotropin-releasing hormone receptor gene structure was ascertained by comparing fragments from autoradiographs in patients and controls. Individual nucleotides were ascertained from DNA sequencing gels. RESULT(S): No GnRH receptor gene deletions or polymorphisms were identified by Southern blot analysis. New restriction-fragment melting polymorphisms using the enzymes DpnII, RsaI, and HaeIII were identified by denaturing gradient gel blots in patients and controls. CONCLUSION(S): Gonadotropin-releasing hormone receptor gene deletions or rearrangements were not observed in our idiopathic hypogonadotropic hypogonadism patients. Denaturing gradient gel electrophoresis failed to identify single-base differences unique to patients with idiopathic hypogonadotropic hypogonadism, dramatically reducing the likelihood that point mutations of the GnRH receptor gene are present in idiopathic hypogonadotropic hypogonadism. PMID- 9418702 TI - Dissociation between intracellular calcium elevation and development of human oocytes treated with calcium ionophore. AB - OBJECTIVE: To develop an acceptable model system to study calcium activation of human oocytes. DESIGN: Study of oocyte development and intracellular calcium [Ca]i dynamics of activated oocytes. SETTING: Research center affiliated with infertility service. MAIN OUTCOME MEASURE: Morphologic evidence of meiotic maturation and cell division under high-power Hoffman optics with an inverted microscope. Meiotic maturation was determined by the number of polar bodies or the presence of a pronucleus, and cell division was determined by evidence of a cleavage furrow or presence of blastomeres. To monitor the effect of calcium ionophore on [Ca]i levels, oocytes were incubated with fura-2 (2 microM) for 30 minutes and [Ca]i was determined by rationing the emission fluorescence (510-nm long-pass filter) during simultaneous excitation at 340 and 380 nm with a microspectrofluorimeter. RESULT(S): All oocytes loaded with fura-2 and then exposed to ionophore exhibited an isolated elevation of [Ca]i, followed by prompt return to baseline levels. None of the oocytes showed signs of cleavage or of meiotic maturation after treatment with calcium ionophore. CONCLUSION(S): Human oocytes activated with calcium ionophore A23187 or ionomycin exhibited elevated [Ca]i but remained resistant to subsequent meiotic maturation and cleavage. Our results differ from some reports of parthenogenetic activation of human oocytes. These differences may result from different activation protocols or culture conditions. Because none of the 126 oocytes cleaved after the activation protocols used in these experiments, this approach should provide an ethically acceptable model system to study calcium dynamics in human oocytes. PMID- 9418703 TI - Subpopulations of human granulosa-luteal cells obtained during early timed and during normally timed follicular aspiration in in-vitro fertilization-embryo transfer cycles. AB - OBJECTIVE: To find the differences between human granulosa-luteal cells obtained during early timed follicular aspiration to prevent severe ovarian hyperstimulation syndrome (OHSS) and during normally timed follicular aspiration. DESIGN: Retrospective analysis of clinical laboratory data. SETTING: In vitro fertilization unit, University Department of Obstetrics and Gynecology, Ljubljana, Slovenia. PATIENT(S): Twenty women undergoing IVF-ET at high risk for OHSS. INTERVENTION(S): Cells were obtained from the follicles of women who were stimulated with hMG and hCG during an early timed follicular aspiration of one ovary, 10-12 hours after hCG, and during a normally timed follicular aspiration of the contralateral ovary, 32-36 hours after hCG administration. MAIN OUTCOME MEASURE(S): Subpopulations of granulosa-luteal cells were observed by computerized image analysis in which hCG was localized using immunoperoxidase staining. RESULT(S): Early timed follicular aspirates contained no oocytes and only a scant number of granulosa cells. Granulosa-luteal cells were smaller than those from normally timed follicular aspirates. We identified three subpopulations in early timed follicular aspirates: nonluteinized, small luteinized, and medium luteinized cells. In normally timed follicular aspirates, four subpopulations were identified, including large luteinized cells. The normally timed follicular aspirates contained more hCG-stained cells. Three staining types of hCG localization were found: on the surface membrane, on the surface membrane and within the cytoplasm, and only within the cytoplasm of cells from normally timed follicular aspirates. Early timed follicular aspirates contained only cells with membrane hCG localization. CONCLUSION(S): We found differences in morphometric characteristics and hCG localization between human granulosa-luteal cells obtained during early timed follicular aspiration to prevent severe OHSS and during normally timed follicular aspiration. PMID- 9418704 TI - Control of human luteal steroidogenesis: role of growth hormone-releasing hormone, vasoactive intestinal peptide, and pituitary adenylate cyclase activating peptide. AB - OBJECTIVE: To examine the possible effect of growth hormone-releasing hormone (GHRH), vasoactive intestinal peptide, and pituitary adenylate cyclase-activating peptide on basal and hCG-stimulated P production by human luteal cells. DESIGN: Cultures of human luteal cells from the early and midluteal phase. SETTING: All corpora lutea were obtained from the Obstetrics and Gynecology Department of the Universita Cattolica, a public care center. PATIENT(S): Ten nonpregnant women between 35 and 47 years of age underwent surgery for various nonendocrine disorders, such as leiomyomatosis. INTERVENTION(S): Corpora lutea were obtained at the time of hysterectomy. MAIN OUTCOME MEASURE(S): Luteal cells were incubated with GHRH, vasoactive intestinal peptide, and pituitary adenylate cyclase activating peptide with or without hCG at different concentrations. RESULT(S): Pituitary adenylate cyclase-activating peptide stimulated P production in a dose- and time-dependent manner, whereas GHRH and vasoactive intestinal peptide did not affect luteal steroidogenesis. None of the three peptides were found to synergize with hCG. CONCLUSION(S): Pituitary adenylate cyclase-activating peptide can influence human luteal steroidogenesis. PMID- 9418705 TI - Decidual leukocyte populations in ectopic pregnancies. AB - OBJECTIVE: To determine whether the recruitment of decidual leukocytes during pregnancy is the same in intrauterine pregnancies (IUPs) versus ectopic pregnancies (EPs). DESIGN: Intrauterine decidual samples from both EPs and IUPs were obtained for the evaluation of leukocyte populations. SETTING: In vitro experiment. PATIENT(S): Women with EPs and women with IUPs. MAIN OUTCOME MEASURE(S): Immunohistochemical identification of decidual leukocyte populations. RESULT(S): We have analyzed the decidual leukocyte populations from three women with EPs by immunohistochemical analysis. The data demonstrate a leukocyte infiltration similar to that found in decidua from normal pregnancies. CONCLUSIONS(S): These data support the hypothesis that decidual leukocyte recruitment and/or increases during pregnancy is primarily hormonally regulated. PMID- 9418706 TI - Charge-based interactions of mammalian sperm with oocytes: inhibition of fertilization of mouse oocytes by ligands of macrophage scavenger receptor(s). AB - OBJECTIVE: To determine whether anionic ligands for the macrophage scavenger receptor inhibit the fertilization of mouse oocytes by mouse spermatozoa. DESIGN: In vitro study of sperm binding and two-cell embryo formation in the presence of scavenger receptor ligands. Sperm-oocyte interaction may be mediated by sulfated sugars. In this study, we tested other nonsulfated anionic ligands for the scavenger receptor for their ability to affect fertilization. The only common feature of these ligands is their anionic nature. SETTING: Oocytes and sperm from mice were used. MAIN OUTCOME MEASURE(S): Binding of sperm to oocytes and subsequent formation of two-cell embryos were determined. RESULT(S): Fucoidin, polyinosinic acid, oxidized low-density lipoprotein, acetyl low-density lipoprotein, and malondialdehyde-modified LDL inhibited the binding and fertilization of mouse sperm to mouse oocytes. Addition of fresh sperm to oocytes previously treated with sperm in the presence of these agents restored the binding and fertilization. CONCLUSION(S): These results show that charge-based interactions analogous to the interactions of the scavenger receptor with its ligands may play an important role in mammalian fertilization. PMID- 9418707 TI - Detection of antisperm antibodies in seminal plasma by flow cytometry: comparison with the indirect immunobead binding test. AB - OBJECTIVE: To compare flow cytometry with the established indirect immunobead binding test (IBT) for the detection of antisperm antibodies in seminal plasma. DESIGN: A prospective, comparative study. SETTING: University-based andrology unit. PATIENT(S): One hundred and fifty-eight men with suspected male factor subfertility. INTERVENTION(S): Seminal plasma samples were incubated with antisperm antibody-negative donor sperm. Surface-bound antibody was detected with fluorescence-labeled antihuman antibody in the flow cytometry assay or with immunobead-labeled antihuman antibody in the IBT. MAIN OUTCOME MEASURE(S): The percentage of sperm that tested positive for surface-bound antibody was determined in the two assays. Seminal plasma was antisperm antibody-positive when > or = 20% of the sperm were antibody-bound, and clinically significant levels were present when > or = 50% of the sperm were antibody-bound. RESULT(S): Of 71 samples that were negative by the IMT, 66 (93%) also were negative by flow cytometry. Of 63 samples that had > or = 50% immunobead binding, 55 had equivalent results by flow cytometry. Overall statistical analysis showed a good correlation between the two assays. CONCLUSION(S): There is a good correlation between the indirect IBT and indirect flow cytometry for the detection of antisperm antibodies in seminal plasma. PMID- 9418708 TI - The ratio between carcinoma antigen-125 levels in the seminal plasma and serum may be a marker for fertilization in intracytoplasmic sperm injection. AB - OBJECTIVE: To assess the relationship between tumor marker carcinoma antigen-125 levels in seminal plasma and serum and fertilization rates in an IVF program, using intracytoplasmic sperm injection (ICSI). DESIGN: A prospective study. SETTING: IVF Unit, Lis Maternity Hospital, Tel Aviv Sourasky Medical Center, Tel Aviv, Israel. PATIENT(S): Twenty-five infertile patients with severe oligo-terato asthenospermia syndrome and 25 fertile male donors. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Serum and seminal plasma carcinoma antigen-125 concentrations and fertilization rate per cycle. RESULT(S): In the infertile group, the seminal plasma carcinoma antigen-125 levels ranged from 22.0 to 1,284.0 U/mL (mean level +/- SD, 229.9 +/- 274.2 U/mL). In the normospermic fertile male donors, the seminal plasma carcinoma antigen-125 concentrations ranged from 12.2 to 336.7 U/mL (mean level +/- SD, 110.1 +/- 91.6 U/mL). This difference was statistically significant. The mean +/- SD ratio between the seminal plasma/serum carcinoma antigen-125 levels differed significantly between the infertile group (47.9 +/- 61.3) and the fertile male donors (5.7 +/- 3.5). In the infertile group, the ratio between the seminal plasma/serum carcinoma antigen-125 levels was found to be negatively correlated with the oocyte fertilization rate. CONCLUSION(S): The ratio between carcinoma antigen-125 levels in the seminal plasma and serum may be an indirect marker for male infertility and fertilization rate in IVF treatment using ICSI. PMID- 9418709 TI - Vasodilator effects of estrogen are not diminished by androgen in postmenopausal women. AB - OBJECTIVE: To compare the effects of estrogen with estrogen-androgen treatment on vaginal blood flow velocity and fingertip postocclusive hyperemic blood flow response. DESIGN: Prospective, randomized, parallel, double-blind study. SETTING: Healthy human volunteers in an academic research environment. PATIENT(S): Postmenopausal women receiving estrogen replacement therapy for at least 12 months and treated with placebo before this investigation. INTERVENTION(S): Esterified estrogens or esterified estrogen + methyltestosterone were administered orally; laser Doppler velocimetry was used to determine vaginal and fingertip blood flow responses at baseline and after 4 and 8 weeks of daily drug administration. MAIN OUTCOME MEASURE(S): Fingertip postocclusive area under curve (AUC); vaginal blood flow velocities. RESULT(S): The AUC for postocclusive fingertip blood flow and vaginal blood flow increased to a greater extent in the estrogen-androgen group, but changes were not statistically significant between groups. CONCLUSION(S): Estrogen-androgen treatment does not diminish the vasodilator effects of estrogen treatment in postmenopausal women. PMID- 9418710 TI - Preimplantation genetic diagnosis increases the implantation rate in human in vitro fertilization by avoiding the transfer of chromosomally abnormal embryos. AB - OBJECTIVE: To verify the percentage of chromosomally abnormal preimplantation embryos in patients with a poor prognosis and possibly to increase the chance of implantation by selecting chromosomally normal embryos. DESIGN: A prospective, randomized, controlled study. SETTING: In vitro fertilization program at the Reproductive Medicine Unit of the Societa Italiana Studi Medicina della Riproduzione, Bologna, Italy. PATIENT(S): In a total of 28 stimulated cycles, the maternal age was > or = 38 years and/or the patient had > or = 3 previous IVF failures, factors that indicated a poor prognosis. After consent, 11 patients underwent preimplantation genetic diagnosis for aneuploidy, whereas 17 controls underwent assisted zona hatching. INTERVENTION(S): Simultaneous analysis of chromosomes X, Y, 13, 18, and 21 in a blastomere biopsied from day-3 embryos. Chromosomal analysis was performed with fluorescence in situ hybridization. Assisted zona hatching was performed on day-3 embryos from the control-group patients. MAIN OUTCOME MEASURE(S): Embryo morphology, results of fluorescence in situ hybridization, clinical pregnancies, and implantation. RESULT(S): In the study group, a total of 61 embryos were analyzed by fluorescence in situ hybridization, and 55% were chromosomally abnormal. Embryo transfer with at least one normal embryo was performed in 10 cycles. Four clinical pregnancies resulted, with a 28.0% implantation rate. In the control group, 41 embryos were transferred in 17 cycles after the assisted zona hatching procedure, yielding four clinical pregnancies and an 11.9% implantation rate. CONCLUSION(S): Infertile patients classified as having a poor prognosis have a high percentage of chromosomally abnormal embryos. The advantage of selecting and transferring embryos with normal fluorescence in situ hybridization results has an immediate impact on implantation. PMID- 9418711 TI - Prospective randomized trial of the effect of two flushing media on oocyte collection and fertilization rates after in vitro fertilization. AB - OBJECTIVE: To assess the value of heparinized saline as a flushing medium for oocyte recovery. DESIGN: Prospective randomized study. SETTING: Academic tertiary referral center for fertility treatment. PATIENT(S): Thirty-five patients, with both ovaries intact having IVF-ET. INTERVENTION(S): Patients were randomized either to have the follicles of the left or right ovary flushed with heparinized normal saline at the time of oocyte recovery for IVF-ET. The contralateral ovary was flushed with heparinized culture medium. Oocytes obtained from each side were cultured separately and assessed for fertilization 18-21 hours after insemination. MAIN OUTCOME MEASURE(S): Collection and fertilization rates. RESULT(S): A total of 481 follicles were aspirated yielding 366 oocytes. Of these, 240 fertilized. From the side flushed with saline 185 oocytes were collected from 237 follicles, which was not significantly different from 181 oocytes collected from 244 follicles on the side flushed with culture medium (odds ratio = 1.23; 95% confidence interval = 0.79-1.92). Similarly, there was no significant difference observed in fertilization rates between oocytes obtained after saline (median 71.4%) and culture medium flush (median 75.0%) (odds ratio = 1.08; 95% confidence interval = 0.68-1.72). CONCLUSION(S): Heparinized normal saline is an equally good but cheaper and more convenient medium than standard heparinized culture medium and could replace it for flushing follicles during oocyte recovery for IVF-ET procedures. PMID- 9418712 TI - Percutaneous vasal sperm aspiration and intrauterine insemination in the treatment of obstructive azoospermia. AB - OBJECTIVE: To treat obstructive azoospermia by using sperm recovered from percutaneous vasal sperm aspiration in IUI. DESIGN: Clinical study. SETTING: Institutional clinic in Jinan. PATIENT(S): Six men with obstructive azoospermia, three of whom were treated with percutaneous vasal sperm aspiration and IUI; sperm recovered from this procedure were used for IUIs. INTERVENTION(S): Spermatozoa used for intrauterine injection were retrieved by percutaneous vasal sperm aspiration and incubated at 37 degrees C for 40 to 60 minutes. MAIN OUTCOME MEASURE(S): Normal pregnancy. RESULT(S): Intrauterine insemination was performed in three patients for one or two cycles, with motile spermatozoa. There was one successful term delivery. CONCLUSION(S): Percutaneous vasal sperm aspiration can be used successfully to recover sperm in men with obstructive azoospermia for use in IUI. The technique is simple and less traumatic than an open surgical procedure. PMID- 9418713 TI - Pregnancy after immature oocyte donation and intracytoplasmic sperm injection. AB - OBJECTIVE: To report a case of pregnancy from in vitro-matured primary oocytes fertilized by ICSI. The pregnancy occurred in a woman who was in an oocyte donation program; the woman's husband had normal sperm parameters. DESIGN: Case report. SETTING: Private general hospital affiliated with a university hospital. PATIENT(S): A recipient with premature ovarian failure, a recipient's husband with normal sperm, and a pregnant woman who donated her oocytes. INTERVENTION(S): Aspiration of immature oocytes during cesarean section, in vitro culture for maturation, ICSI of matured oocytes, coculture of fertilized oocytes. MAIN OUTCOME MEASURE(S): Fertilization of oocytes by ICSI, and cleavage of embryos by Vero cell coculture. RESULT(S): Two of seven immature oocytes became metaphase II oocytes, and both were fertilized by ICSI. The two zygotes were cocultured on Vero cells to become grade 1 two-cell embryos. Pregnancy was obtained after transfer. CONCLUSION(S): More studies are necessary to clarify whether ICSI can increase the fertilization rate of in vitro-matured primary oocytes, and to clarify the role of coculture in fertilization. PMID- 9418714 TI - Hysteroscopic treatment of intrauterine adhesions is safe and effective in the restoration of normal menstruation and fertility. AB - OBJECTIVE: To assess the safety and efficacy of hysteroscopic adhesiolysis in patients with recurrent pregnancy loss and infertility. DESIGN: Retrospective case report series. SETTING: The obstetrics and gynecology clinic of a medical school. PATIENT(S): Forty women with recurrent pregnancy loss or infertility resulting from intrauterine adhesions. INTERVENTION(S): Hysteroscopic adhesiolysis in patients with recurrent pregnancy loss and infertility. MAIN OUTCOME MEASURE(S): Postoperative adhesion formation, intraoperative complication, conceivement after surgery, pregnancy rate, and pregnancies resulted in term or viable preterm infants. RESULT(S): Most patients with minimal or moderate adhesions were free of adhesions when compared with postoperative control subjects. However, adhesion re-formation was noted in 60% of the patients who initially had severe adhesions. Normal menstrual flow was restored in 81% of the patients. All the patients who had recurrent pregnancy loss conceived after treatment, and 71% of the pregnancies resulted in a term or viable preterm infant. Of the 16 infertile patients treated, 10 (63%) conceived and 6 (37%) were delivered of viable infants. CONCLUSION(S): Hysteroscopic adhesiolysis is a safe and effective procedure for restoring the normal menstrual pattern and fertility. The initial severity of the adhesions appears to correlate best with the reproductive outcome. PMID- 9418715 TI - Fertilization and pregnancy after assisted oocyte activation and intracytoplasmic sperm injection in a case of round-headed sperm associated with deficient oocyte activation capacity. AB - OBJECTIVE: To investigate a method of assisted activation of human oocytes for the treatment of infertility resulting from globozoospermia associated with deficient oocyte activation capacity. DESIGN: The mouse oocyte activation test was used to analyze the oocyte activation capacity of the sperm cells of a patient with globozoospermia. Fresh donor human oocytes were used for determining the most appropriate procedure for oocyte activation. SETTING: Infertility Center, University Hospital of Ghent. PATIENT(S): A couple with infertility resulting from globozoospermia. INTERVENTION(S): Intracytoplasmic sperm injection, assisted oocyte activation, and embryo transfer. MAIN OUTCOME MEASURE(S): Oocyte activation and fertilization rates, implantation, and pregnancy. RESULT(S): Deficiency in oocyte activation capacity was found in the sperm of a patient with globozoospermia. This deficiency was proven by the mouse oocyte activation test and was confirmed further by lack of activation of human oocytes after simple sperm injection. Only human oocytes that were injected with sperm and subjected to calcium chloride and ionophore treatment underwent activation. Transfer of embryos obtained by this procedure of assisted oocyte activation resulted in an ongoing pregnancy. CONCLUSION(S): Assisted oocyte activation of human oocytes is useful when globozoospermia is associated with absence of oocyte activation capacity in the sperm. These cases can be identified by the mouse oocyte activation test. PMID- 9418716 TI - Follicle aspiration, sperm injection, and assisted rupture (FASIAR): a simple new assisted reproductive technique. AB - OBJECTIVE: To describe the first successful application of a new fertility enhancing technique. DESIGN: Case report. SETTING: Academic fertility program. PATIENT(S): A 36-year-old nulligravid woman undergoing inseminations with frozen thawed donor sperm. INTERVENTION(S): Ovarian superovulation, follicle aspiration, sperm injection, and assisted follicular rupture. MAIN OUTCOME MEASURE(S): Assessment of feasibility of technique and pregnancy outcome. RESULT(S): After failing to conceive during 16 cycles of IUI, the patient successfully achieved an ongoing pregnancy during the second follicle aspiration, sperm injection, and assisted rupture (FASIAR) attempt. CONCLUSION(S): Follicle aspiration, sperm injection, and assisted rupture combines the concepts of superovulation, IUI, and peritoneal oocyte and sperm transfer to obviate the possibility of luteinized unruptured follicle syndrome, assist oocyte release, and ensure gamete intermixing. It also can be used to reduce the number of ovulating oocytes and thus to reduce the risk of multiple gestations. Follicle aspiration, sperm injection, and assisted rupture is a new, simple, office-based procedure that does not require embryologic expertise beyond sperm preparation as for IUI, yet promises to be more successful than IUI. PMID- 9418717 TI - Do "sponsoring embryos" really respond? PMID- 9418718 TI - Protocols and IRB approval for post-menopausal pregnancies. PMID- 9418719 TI - Protocols and IRB approval for post-menopausal pregnancies. PMID- 9418720 TI - Vision research in the 21st century. PMID- 9418721 TI - Adenosine-induced relaxation of cultured bovine retinal pericytes. AB - PURPOSE: To investigate the effect of adenosine on the contractile tone of cultured bovine retinal pericytes. METHODS: Changes in the contractile tone were quantified as the changes in the summed length of wrinkles induced by pericytes on the silicone surface on which the cells were grown. RESULTS: Adenosine at 10( 9) M had no effect. In the range of 10(-8) to 10(-4) M, adenosine caused relaxation of pericytes in a concentration-dependent manner. Complete relaxation was induced by 10(-5) M to 10(-4) M adenosine. The concentration of adenosine that produced 50% relaxation was 3 x 10(-7) M. At all concentrations, relaxation began within 1 minute, reached the maximum within 5 to 10 minutes, and persisted for at least 30 minutes. After a washout of 3 x 10(-7) M adenosine, the reduced contractile tone recovered to the original level in 10 minutes. The adenosine induced relaxation (3 x 10(-7) M) was completely abolished in the presence of 8 phenyl theophylline (10(-5) M), a nonselective adenosine receptor antagonist. The selective A1 receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX) at 10(-6) M did not reduce the effect of adenosine (3 x 10(-7) M). Conversely, the selective A2 receptor antagonist CP-66,713 at 10(-8) M partially inhibited (and at 10(-7) M, completely inhibited) the relaxation induced by adenosine (3 x 10( 7) M). The adenosine receptor antagonists-8-phenyl theophylline (10(-5) M), DPCPX (10(-6) M), and CP-66,713 (10(-7) M) by themselves had no effect on the contractile tone of pericytes. CONCLUSIONS: Adenosine causes relaxation of pericytes through the activation of the adenosine A2 receptor. Adenosine, which accumulates under ischemic conditions, may help to regulate local capillary blood flow. PMID- 9418722 TI - Feasibility of laser-targeted photoocclusion of the choriocapillary layer in rats. AB - PURPOSE: A new method, laser-targeted photoocclusion, was developed to occlude choroidal neovascularization while minimizing damage to the overlying retina. The ability to occlude normal choriocapillary layer in rats was evaluated as a first test of the feasibility of treating choroidal neovascularization with this method. METHOD: A photosensitive agent, aluminum phthalocyanine tetrasulfonate, encapsulated in heat-sensitive liposomes, was administered intravenously along with carboxyfluorescein liposomes. A low-power argon laser (retinal power density of 5.7 W/cm2) locally released a photosensitizer bolus, monitored by the simultaneous release of carboxyfluorescein. A diode laser (operating at 675 nm with a retinal power density of 0.27 W/cm2) activated the photosensitizer with its release. RESULTS: Vessels in the choriocapillary layer were occluded at day 3 after laser treatment and remained unchanged during the 30-day follow-up. Larger choroidal vessels and retinal capillaries remained perfused. Control experiments excluded possible effects of heat or activation of free photosensitizer. Pilot histologic studies showed no damage to the retinal pigment epithelium. CONCLUSIONS: Laser-targeted photoocclusion caused selective occlusion of normal choriocapillaries while sparing overlying retinal pigment epithelium and retinal vessels. The method has potential as a treatment of choroidal neovascularization that may minimize iatrogenic loss of vision. PMID- 9418723 TI - Specific retinal diacylglycerol and protein kinase C beta isoform modulation mimics abnormal retinal hemodynamics in diabetic rats. AB - PURPOSE: Elevation of diacylglycerol (DAG) and protein kinase C (PKC) levels in diabetic vascular tissue is associated with abnormalities of retinal and renal hemodynamics. The object of this study was to determine whether direct elevation of retinal DAG levels, in the absence of diabetes or hyperglycemia, can mimic the hemodynamic abnormalities normally observed in diabetic rats. Retinal DAG levels were elevated using an inhibitor of DAG kinase that converts DAG to phosphatidic acid. The effectiveness of a specific PKC-beta isoform inhibitor introduced directly into the retinas of diabetic rats in reversing diabetes-related abnormal retinal hemodynamics was also investigated. METHODS: For retinal blood flow studies, diacylglycerol kinase (DGK) inhibitor R59949, at various concentrations, was injected into the vitreous of nondiabetic Sprague-Dawley rats (n = 33), and a PKC-beta isoform-selective inhibitor LY333531 was injected into the vitreous of rats with streptozotocin (STZ)-induced diabetes of 2 weeks' duration (n = 21). Retinal hemodynamic changes were quantitated using video-based fluorescein angiography. Total DAG levels were assayed from five nondiabetic rat retinas after DGK inhibition and retinal PKC activities were assayed from six diabetic rat retinas after PKC-beta inhibition. RESULTS: DGK inhibitor R59949 injected into the vitreous dose dependently increased the mean circulation time (MCT) and decreased retinal blood flow (EC50 = 10(-8) M). After 30 minutes, 10(-5) M R59949 induced a 1.7-fold increase in total retinal DAG levels, compared with the levels in vehicle-injected eyes, an increase in MCT from 0.87 +/- 0.05 seconds to 1.44 +/- 0.12 seconds (P < 0.01) and a decrease in retinal blood flow from 105.3 +/- 6.5 pixel2/second to 64.1 +/- 5 pixel2/second (P < 0.01). The effect of R59949 was sustained for 60 minutes after injection. These retinal hemodynamic parameters after DGK inhibition were comparable to those measured at baseline in rats with STZ-induced diabetes of 2 weeks' duration (MCT = 1.38 +/- 0.20 seconds; retinal blood flow = 68 +/- 11.2 pixel2/second). Intravitreal injection of the PKC-beta inhibitor (LY333531) at 10(-5) M in diabetic rats decreased by a factor of 1.6 the diabetes-related increased PKC activation, decreased the prolonged MCT (0.98 +/- 0.13 seconds; P < 0.01) and increased retinal blood flow (93.4 +/- 14.2 pixel2/second; P < 0.01). The measured retinal circulatory parameters after PKC inhibition in the retina were comparable to those measured at baseline in the nondiabetic rats. CONCLUSIONS: These results provide direct evidence that DAG elevation and subsequent PKC-beta isoform activation are the primary biochemical sequelae responsible for the development of the abnormal retinal hemodynamics observed in diabetic rats. PMID- 9418724 TI - Fluorophotometric quantitation of oxidative stress in the retina in vivo. AB - PURPOSE: To establish a new fluorophotometric method to quantitate oxidative stress in the retina in vivo with a hydrogen peroxide (H2O2)-sensitive fluorescent dye. METHODS: For in vitro fluorophotometric study, nonfluorescent 2',7'-dichlorofluorescein (DCFH) was incubated with H2O2 (10 pM to 100 nM), and the production of fluorescent 2',7'-dichlorofluorescein (DCF) was measured with fluorophotometric analysis. The inhibitory effect of catalase was also examined. For in vivo fluorophotometric study, rabbit eyes received vitrectomy and were perfused with 5 microM 2',7'-dichlorofluorescein diacetate (DCF-DA) or 2',7' dichlorofluorescein diacetate (DCFH-DA). For oxidative stress, 300 microM H2O2 was infused after perfusion of DCFH-DA. Fluorophotometric measurements of the chorioretinal peak were performed. The eyes were enucleated for fluorescent microscopic examination to determine the localization of DCF fluorescence. RESULTS: H2O2 converted DCFH to DCF in a dose-dependent manner, which was inhibited by catalase dose dependently. In vivo fluorophotometric study showed DCF-DA and DCFH-DA caused production of 2006 +/- 274 picomole/ml (mean +/- SD, n = 5) and 8.35 +/- 1.11 picomole/ml (n = 5), respectively, in the chorioretinal peak. DCFH-DA with stimulation by H2O2 induced 30.7 +/- 13.1 (n = 4) picomole/ml DCF. Fluorescent microscopy showed DCF production in the retina was significant in the eye treated with DCF-DA and minimal in the eye treated with DCFH-DA. Moderate DCF production in the nerve fiber layer was observed in the eye treated with DCFH-DA and H2O2. CONCLUSIONS: This new fluorophotometric method with DCFH DA may be useful in quantitatively evaluating oxidative stress in the retina in vivo. PMID- 9418725 TI - Vascular endothelial growth factor and vascular permeability changes in human diabetic retinopathy. AB - PURPOSE: The authors used histochemical analysis to determine whether increased vascular endothelial growth factor (VEGF) immunoreactivity in diabetic retinal vessels is related to increased vascular permeability, as indicated by human serum albumin (HSA) immunostaining, or to presumed retinal hypoxia as demonstrated by decreased vascularity. A correlation between VEGF and HSA in cryosections with angiopathic changes in the adenosine diphosphatase (ADPase) flat-embedded fellow retinas was sought. Because VEGF is a heparin-binding protein, the relation between VEGF and heparan sulfate proteoglycan (HSPG) immunoreactivities was also investigated. METHODS: Cryopreserved eyes from 18 diabetic and 9 nondiabetic subjects removed after death were sectioned and immunohistochemical analysis was performed with antibodies against VEGF, HSA, HSPG, vWf (von Willebrand factor), and collagen IV. The fellow retinas were prepared by our ADPase flat-embedding technique to determine the degree of diabetic retinopathy. The number of positive vessels for each antibody and antibody localizations were determined by light microscopy. RESULTS: The average number of VEGF-stained vessels in diabetic retinas was significantly higher than in nondiabetic retinas (P = 0.04). In diabetic retinas, there was a positive correlation between the distribution of VEGF-positive vessels and the distribution of HSA- and HSPG-positive vessels. No such correlation was observed in nondiabetic eyes. In many cases, HSPG immunoreactivity appeared colocalized with VEGF immunoreactivity, suggesting VEGF binding to HSPG. The comparison with the ADPase flat-embedded fellow retinas suggested that increased VEGF immunoreactivity and vascular permeability may occur before morphologic changes in the vasculature. CONCLUSIONS: Vascular endothelial growth factor immunoreactivity was correlated with increased vascular permeability to macromolecules and appears to be increased in diabetic subjects before the onset of retinopathy. PMID- 9418726 TI - The effect of acute experimental retinal vein occlusion on cat retinal vein pressures. AB - PURPOSE: Retinal ischemic damage associated with retinal vein occlusion is exacerbated by fluid extravasation and hemorrhage, which may be caused by increased permeability, elevated intravascular pressure, or both. Direct measurement of the retinal vein pressure in the cat after acute experimental retinal vein occlusion may define the role of intravascular pressures in fluid extravasation associated with this condition. METHODS: Intravenous retinal pressure measurements were obtained using a micropipette connected to a servonull device and positioned by a robot micromanipulator, while a major retinal vein near the optic disc was occluded by argon laser radiation delivered through an optical fiber positioned by a manual micromanipulator. After occlusion, retinal vein pressures were measured on both sides of the occlusion site at a controlled intraocular pressure of 20 mm Hg. RESULTS: Upstream of the occlusion site, the retinal vein pressures were not greatly elevated, although they were significantly different from controls. Downstream vein pressures were significantly lower than controls, but vascular collapse near the optic nerve was not observed. CONCLUSIONS: In retinal vein occlusion, venous pressures in a segmental retinal circulatory bed are not substantially elevated, thus implying the presence of a pressure-release mechanism and implicating vascular damage for the increased transvascular fluid flux. The lack of vascular collapse downstream of the occlusion site suggests collateral communication before a large intraocular pressure-dependent resistance segment that lies between the intraocular and extraocular vessels. PMID- 9418727 TI - Localization of CSNBX (CSNB4) between the retinitis pigmentosa loci RP2 and RP3 on proximal Xp. AB - PURPOSE: Proximal Xp harbors many inherited retinal disorders, including retinitis pigmentosa (RP) and congenital stationary night blindness, both of which display genetic heterogeneity. X-linked congenital stationary night blindness (CSNBX) is a nonprogressive disease causing night blindness and reduced visual acuity. Distinct genetic loci have been reported for CSNBX at Xp21.1, which is potentially allelic with the RP3 gene, and at Xp11.23, which is potentially allelic with the RP2 gene. The study to identify the RP2 gene led to an extended study of families with potentially allelic diseases that include CSNBX. METHODS: Haplotype analysis of a family diagnosed with CSNBX was performed with 17 polymorphic markers on proximal Xp covering previously identified loci for CSNBX and XLRP. Two-point and multipoint lod scores were calculated. RESULTS: Informative recombinations in this family define a locus for CSNBX (CSNB4) with flanking markers DXS556 and DXS8080 on Xp11.4 to Xp11.3, an interval spanning approximately 5 to 6 cM. A maximum lod score of 3.2 was calculated for the locus order DXS556-1 cM-(CSNB4-DXS993)-2 cM-DXS1201. CONCLUSIONS: The results describe a new localization for CSNBX (CSNB4) between the RP2 and RP3 loci on proximal Xp. CSNB4 is not allelic with any previously reported XLRP loci; however, the interval overlaps the locus reported to contain the cone dystrophy (COD1) gene, and both diseases are nonrecombinant with DXS993. Because mutations in the RPGR gene to date account for disease in only a small proportion of RP3 families, the possibility that this new locus (CSNB4) also segregates with an as yet unidentified XLRP locus cannot be excluded. PMID- 9418728 TI - Familial subepithelial corneal amyloidosis--a lactoferrin-related amyloidosis. AB - PURPOSE: To isolate the protein that collects in increased amounts beneath the corneal epithelium in familial subepithelial corneal amyloidosis (FSCA), also known as gelatinous droplike corneal dystrophy, and to identify it by N-terminal amino acid sequencing. METHODS: Peptides resulting from pepsin digestion of a unique protein isolated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis from frozen tissue from two corneas with FSCA were purified by high-pressure liquid chromatography followed by protein sequence analysis. The protein was identified by amino acid sequencing, Western blotting, and immunohistochemistry. RESULTS: A protein was identified in two corneas with FSCA that was not present in normal corneas or in corneas with other disorders. The amino acid sequences of two peptides derived from this protein were identical to portions of lactoferrin. The unique protein reacted with rabbit antihuman lactoferrin after Western blotting. The presence of lactoferrin in the amyloid within affected corneas was confirmed using the immunoperoxidase method on formalin-fixed, paraffin-embedded tissue sections and lactoferrin antiserum. CONCLUSIONS: Corneal tissue with FSCA contains lactoferrin, and this is the first form of amyloidosis found to be associated with this protein. Because lactoferrin is a product of lacrimal glands, the corneal lactoferrin may be derived from the tears. Because the gene for lactoferrin is on chromosome 3 (3q21-q23), this locus is a potential site for the FSCA gene. PMID- 9418729 TI - Extralenticular expression of Xenopus laevis alpha-, beta-, and gamma-crystallin genes. AB - PURPOSE: Extralenticular expression of alpha- and beta-crystallin genes has been demonstrated in mammals and expression of gamma-crystallin genes has been shown in Xenopus laevis. To determine a possible correlation between lens determination and crystallin gene expression, the site of expression of (a member of) the alpha , beta-, and gamma-crystallin gene families was observed before and during lens formation in X. laevis. METHODS: The partial complementary DNAs (cDNAs) of alpha A- and beta A4-crystallin and a gamma-crystallin were cloned from an X. laevis lens cDNA library. The corresponding antisense RNAs were used to analyze the expression of these genes during X. laevis development by wholemount in situ hybridization. RESULTS: Expression of the beta A4- and gamma-crystallin (but not alpha-crystallin) genes could first be detected in the animal cap of the X. laevis gastrula. The beta A4- and gamma-crystallin messengers were also found in the first stage of lens development, when the ectodermal tissue overlying the optic vesicle thickens to form the lens placode. alpha A-crystallin messenger RNAs were only detectable when the lens epithelial cells were formed. CONCLUSIONS: In contrast to observations in most vertebrates, expression of the beta A4- and gamma-crystallin genes was observed to precede that of the alpha A crystallin gene during lens development of X. laevis, reflecting the determination that in amphibians, the (presumptive) fiber cells are formed before the epithelial cells, whereas in vertebrates, the order is reversed. Expression of beta A4- and gamma-crystallin genes in the ectodermal tissue of the X. laevis gastrula shows that these genes are expressed when this tissue gains competence for lens formation. PMID- 9418730 TI - Prostaglandins increase matrix metalloproteinase release from human ciliary smooth muscle cells. AB - PURPOSE: To identify matrix metalloproteinases (MMPs) released by ciliary smooth muscle cells in vitro and to determine whether MMP release is altered by exposure to prostaglandins (PGs). METHODS: Human ciliary smooth muscle cells were grown to confluence in monolayer cultures and treated with PGF2 alpha, 11-deoxy-PGE1, or PhXA85 (the nonesterified analogue of PhXA41) for 12 to 72 hours. The activity of MMP in the medium was assayed using gelatin and casein zymography. Identification of the specific MMP associated with each band was made by Western blot analysis. Band intensity, which reflects activity, was measured with a scanning laser densitometer. RESULTS: Three major bands appeared in the gelatin zymographs at positions corresponding to molecular weights of 62 kDa, 68 kDa, and 97 kDa. A single band at 50 kDa predominated in the casein zymograms. Substitution of EDTA for calcium and zinc in the development solution eliminated the appearance of these bands, indicating that they reflect MMP activity. Immunoblotting, using MMP specific antibodies, confirmed that the three bands in the gelatin zymographs were MMP-1, MMP-2, and MMP-9, respectively; the single band in the casein zymographs was MMP-3. Treatment with 200 nM PGF2 alpha, 11-deoxy-PGE1, or PhXA85 for 72 hours increased the combined density scores for MMP-1 and MMP-2 by 37%, 64%, and 27%; the density scores for MMP-9 by 268%, 253%, and 125%; and the density scores for MMP-3 by 35%, 71%, and 22%, respectively. CONCLUSIONS: These results indicate that ciliary smooth muscle cells can secrete MMP-1, MMP-2, MMP 3, and MMP-9. In addition, exposure to PGF2 alpha, 11-deoxy-PGE1, or PhXA85 increases production of all four MMPs. These observations support the hypothesis that increased MMP production by ciliary muscle cells has a role in increasing uveoscleral outflow facility after topical PG administration. PMID- 9418731 TI - Vasoactive intestinal peptide stimulation of human trabecular meshwork cell growth. AB - PURPOSE: To demonstrate that vasoactive intestinal peptide (VIP), a 28-amino acid neuropeptide, is a growth factor of human trabecular meshwork (TM) cells in culture and in a corneoscleral explant organ culture treated with laser trabeculoplasty (LTP). METHODS: Proliferating human TM cells in cell cultures were incubated with VIP for 20 hours, followed by total cell number determination, using a Coulter counter. The percentage of proliferating TM cells was assessed, using an antibody against the proliferating cell nuclear antigen (PCNA). To test the growth effect of VIP on TM cells in situ, corneoscleral explants in organ cultures were first treated with argon LTP to initiate TM-cell proliferation and then were exposed to VIP for 48 hours. The mitotic TM cells were demonstrated immunocytochemically, using anti-PCNA in paraffin sections of the explants; and the total number of TM cells was determined after paraffin sections were counterstained by hematoxylin. RESULTS: Vasoactive intestinal peptide dose-dependently stimulated the proliferation of TM cells in cell culture. Treatment with 5 x 10(-10) M VIP resulted in a maximal increase of 40% in cell number. The effect of VIP was blocked by a VIP antagonist. The number of PCNA-stained TM cells and the total cell number in the TM in LTP-treated corneoscleral explants were increased by VIP. CONCLUSIONS: Exogenously applied VIP stimulated the proliferation of human TM cells in subconfluent cultures and in LTP-treated corneoscleral explants. In that LTP has been shown to increase the number of TM cells in situ, the growth stimulatory effect of VIP may help enhance this therapy. PMID- 9418732 TI - The effect of cytochalasin D on outflow facility and the trabecular meshwork of the human eye in perfusion organ culture. AB - PURPOSE: To determine the effect of cytochalasin D on outflow facility in the human anterior segment, and the histologic changes that accompany the effect. METHODS: Human anterior segments were studied in perfusion organ culture. The anterior segment from one eye received cytochalasin D, and that from the fellow control eye received vehicle; doses ranged from 0.06 mg/ml to 27.7 mg/ml. The duration of action and the effect of repeated doses were studied, and the accompanying histologic changes were assessed in 12 pairs of anterior segments. RESULTS: Cytochalasin D in concentrations of 0.6 mg/ml and 1.1 mg/ml caused increases in outflow facility of 42% and 37%, respectively (P < 0.05), with a peak effect 2 to 6 hours after infusion and a duration of action of approximately 14 hours. Anterior segments were not responsive to repeated doses (24 hours apart). Compared with the effect of vehicle in control anterior segments, cytochalasin D caused scattered breaks in the inner wall endothelial lining of Schlemm's canal (4.6 +/- 2.5% versus 0.7 +/- 0.6%; P = 0.02; anterior segments fixed during maximum drug effect). No increase in the amount of optically empty space within the juxtacanalicular tissue was seen. Inner wall breaks persisted, even in eyes in which the outflow facility had returned to baseline; the basement membrane and subendothelial matrix of the inner wall remained intact. Final intraocular pressure was inversely correlated with the length of optically empty space immediately adjacent to the inner wall. CONCLUSIONS: Cytochalasin D can increase outflow facility in the anterior segment of the human eye and causes ruptures of the inner wall of Schlemm's canal. These breaks persist, even when interocular pressure returns to baseline; the basement membrane and subendothelial matrix of the inner wall appear to remain intact. The final intraocular pressure was inversely correlated with the length of optically empty space immediately adjacent to the inner wall. PMID- 9418733 TI - Poor stereopsis can support size constancy in albinism. AB - PURPOSE: The size of a retinal image is inversely related to the distance to the object that generates the image. Normal subjects therefore exhibit size constancy, in which the perceived size of an image is scaled according to its perceived distance. Albinos usually have such poor binocular vision that they perform very poorly on clinical tests for stereopsis. To investigate the functional consequences of this poor stereopsis, we investigated whether stereopsis in these subjects could support size constancy. METHODS: The stereothresholds of 10 albinos and 12 normal control subjects were measured. The presence of absence of size constancy was investigated by having subjects equate the subjective size of stereoscopically presented images whose image disparity indicated that they were at different distances. RESULTS: Laboratory results indicated that eight albinos (including five whose clinical tests indicated a lack of stereopsis) had measurable stereopsis of several thousand are seconds or better. Of these, four also exhibited size constancy. CONCLUSIONS: Albinos who do not demonstrate stereopsis on clinical tests can have stereoscopic perception that commonly used clinical tests do not detect. Moreover, some of these patients even use this poor stereopsis in judging the size of stereoscopically presented images. PMID- 9418734 TI - Decrease in saccadic performance after many visually guided saccadic eye movements in monkeys. AB - PURPOSE: To investigate the influence of repeated saccades and of background illumination on the metrics and dynamics of visually guided targeting saccades. METHODS: Eye movements were measured by magnetic search coil technique in seven trained monkeys (Macaca mulatta) while they performed many visually guided saccades in the dark or in dim background light. RESULTS: After 2000 to 7000 saccades in the dark, peak eye velocity on the average decreased by 20%, saccadic gain decreased slightly by 4.5%, and saccadic latency increased by 15%. All parameters also showed increased variability. In contrast, when testing was done in dim light, there was little to no change in average saccadic metrics and latency. CONCLUSIONS: The changes in saccadic metrics and dynamics in the dark do not reflect a change of the ocular plant but may reflect a change in the cortical or cerebellar influences on the brain stem burst generator linked to the monkeys attentional state. Background light mostly prevents this change. PMID- 9418735 TI - Extraocular fast myosin heavy chain expression in the levator palpebrae and retractor bulbi muscles. AB - PURPOSE: To determine whether the levator palpebrae superioris (LPS) and the retractor bulbi (RB) muscles, which share contractile characteristics with extraocular muscles (EOMs), express fast EOM-specific myosin heavy chain (MyHC) in the rabbit and other mammalian species. METHODS: Cryostat sections of rabbit eye and limb muscles were stained by indirect peroxidase immunohistochemical procedures using monoclonal antibodies (MAbs), including one (4A6) against EOM specific fast MyHC. Myosin heavy chain isoforms from these muscles were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and the 4A6-reactive component was identified by immunoblotting. RESULTS: MAb 4A6 stained muscle fibers in rabbit LPS and RB. SDS-PAGE resolved a rabbit EOM-specific MyHC isoform (band 1) from two other components (bands 2 and 3) that comigrated with limb fast MyHCs. MAb 4A6 reacted only with band 1. Rabbit LPS and RB also displayed corresponding MyHC components with the same mobilities and immunoreactivities as bands 1 to 3 in the EOM. MAb 4A6 also stained muscle fibers in monkey and cat LPS, but it failed to stain muscle fibers in cat RB and rat LPS and RB. CONCLUSIONS: The expression of EOM-specific fast MyHC in EOM, LPS, and RB reflect their common developmental origin and similar contractile characteristics. These properties set them apart from other skeletal muscle groups. Eye muscles may constitute a distinct muscle group or allotype characterized by unique properties, including their propensity or resistance to disease. PMID- 9418736 TI - Confocal microscopy of human lens membranes in aged normal and nuclear cataracts. AB - PURPOSE: To visualize the structure and determine the continuity of lipid membranes in lens fiber cells (LFCs) from human aged normal and cataractous lenses. METHODS: Thick sections from human nuclear cataracts and aged normal lenses were stained with the lipophilic probe DiI, and then analyzed by confocal microscopy. Staining patterns of membranes were observed in individual optical sections or three-dimensional projections of z-series taken in longitudinal section and cross-section of LFCs from different regions within the lens nucleus. RESULTS: DiI bound to and delineated the plasma membrane of LFCs from all regions of the lens nucleus. Three-dimensional projections of z-series from aged normal and cataractous lenses suggested that some of the stained lipid membranes were not continuous with LFC plasma membrane of cataractous lenses. CONCLUSIONS: The results obtained using these methods demonstrated that lipid membranes, discontinuous with the plasma membrane of LFCs, were indicative of a novel process occurring predominately in cataractous human lenses. PMID- 9418738 TI - Multifocal electroretinogram in myopia. AB - PURPOSE: To investigate early changes of retinal function in myopic eyes, patients with various degrees of myopia underwent multifocal electroretinography (multifocal ERG). METHODS: Thirty young patients (mean age, 26.1 years) were divided into three groups according to the degree of refractive error: emmetropia/low, medium, and high myopia. All groups were matched by sex and age. All subjects had good corrected visual acuities of better than 1.0 and normal color vision. No other ophthalmoscopic abnormalities except for tessellated fundus existed. Three grouping programs (all-traces, rings, and quadrants) were used, and all-traces grouping waveforms, six-ring grouping waveforms, and four quadrant grouping waveforms were determined from 103 local responses. We studied the differences of amplitudes and latencies of the waves among the myopic groups and their relation to refractive error and axial length. RESULTS: Amplitudes were reduced and latencies were delayed as the refractive errors increased. The amplitudes in the peripheral areas were more reduced in the myopic groups. CONCLUSIONS: The reduced amplitude and delayed latency of each wave of multifocal ERG in myopia resulted primarily from cone function loss, which may occur even when the only noted retinal change is tessellated fundus. PMID- 9418737 TI - Induction of anterior chamber-associated immune deviation by corneal allografts placed in the anterior chamber. AB - PURPOSE: Although mice with long-accepted orthotopic corneal allografts display donor-specific anterior chamber-associated immune deviation (ACAID), this deviant response is not detected until well after the fate of the grafts is decided. To determine the efficiency with which corneal tissue itself can induce ACAID, allogeneic corneal segments were inserted into anterior chambers (AC) of normal mouse eyes. METHODS: Central corneas from normal eyes of C57BL/6 (allogeneic) and BALB/c (syngeneic) donors were cut into wedge-shaped fragments measuring approximately 0.3 x 2.0 mm (in some experiments the corneal epithelium was removed) and inserted into the AC adjacent to recipient endothelium. Recipients of these fragments were evaluated for donor-specific delayed hypersensitivity (DH) and ACAID, and fragment-containing eyes were tested for their capacity to support ACAID to an irrelevant antigen. RESULTS: Syngeneic and allogeneic corneas survived indefinitely in the AC without evidence of inflammation or rejection. Although fragments of cornea (with or without epithelial layers) placed at extraocular sites were potent inducers of DH, within the eye only epithelium bearing grafts induced DH. Moreover, this DH response was short-lived. Recipients of allogeneic corneal fragments in the AC developed ACAID by 8 weeks, but not at 1 week. Moreover, fragment-containing eyes supported ACAID induction when bovine serum albumin was injected into the AC. CONCLUSIONS: Anterior chamber-associated immune deviation can be induced by allogeneic corneal tissue inserted into the AC, but its onset is delayed. The delay may be dictated by persistence of donor epithelium on the graft, which promotes DH. Once the epithelium is lost, DH disappears and ACAID emerges. Anterior chamber-associated immune deviation may contribute to the maintenance of corneal allograft viability. PMID- 9418739 TI - Expression and upregulation of microtubule-associated protein 1B in cultured retinal pigment epithelial cells. AB - PURPOSE: During routine cell culture and under pathologic conditions, human retinal pigment epithelial (RPE) cells lose epithelial characteristics and change their morphology. In this study, changes in gene expression in RPE cells of different generations were evaluated by polymerase-chain-reaction-based differential display mRNA analysis (DD-RT-PCR). METHODS: Total RNA was prepared from freshly isolated and cultured human RPE cells of passages P0 and P3 and was subjected to DD-RT-PCR. One band with enhanced expression was excised, reamplified, and partially sequenced, using a modified dideoxy chain termination approach. Expression of the corresponding protein was ascertained by immunocytochemical analysis. RESULTS: Differential display RT-PCR showed enhanced expression of a specific RNA in P3 cells compared with that in P0 cells. Sequence alignment revealed 98% identity with the 3' end of the coding sequence of human microtubule-associated protein 1B (MAP1B). Confirmation of induced expression of MAP1B mRNA was obtained by PCR with specific primers and by immunocytochemical analysis in cultured RPE cells and in surgically removed epiretinal membranes from patients with proliferative vitreoretinopathy. No expression of MAP1B mRNA or protein was detected in freshly isolated RPE cells. CONCLUSIONS: Differential display RT-PCR in RPE cells with subsequent sequence analysis allows characterization of the maturation- and differentiation-dependent expression of previously undetected genes and gene products in cultured RPE cells. PMID- 9418740 TI - Real-time, noninvasive in vivo assessment of adeno-associated virus-mediated retinal transduction. AB - PURPOSE: To evaluate the efficiency, cell specificity, stability, and toxicity of recombinant adeno-associated virus (rAAV)-mediated retinal transduction in vivo in the adult immunocompetent mouse. To assess the usefulness of green fluorescent protein (GFP) for real-time, noninvasive monitoring of retinal transgene expression in vivo. METHODS: Assessment of ocular GFP expression was performed in cohorts of mice for 11 weeks after subretinal injection of a recombinant adeno associated virus carrying the complementary DNA (cDNA) for GFP. Examinations were performed in vivo by direct observation of fluorescence by ophthalmoscopy, using excitation-barrier filters. Histologic analyses of retinal tissue were used to identify transduced cells and to assess inflammation. RESULTS: Retinal GFP expression can be monitored in vivo using real-time, noninvasive imaging. Recombinant AAV efficiently transduces a variety of cells of the neural retina and of the retinal pigment epithelium (RPE). Transgene expression was not observed until 1 week after infection. The number of GFP-expressing cells increased over 3 weeks, and expressing photoreceptors and RPE, cells persisted at least through 11 weeks (the termination of the experiment). There was no clinical or histologic evidence of inflammatory response. CONCLUSIONS: Retinal gene transfer mediated by rAAV is stable and efficient and is associated with no clinically or histologically detectable toxicity or immune reaction. Green fluorescent protein allows noninvasive assessment of the extent and location of retinal transgene expression as a function of time and promises to be useful alone and as a tag for other transgenes delivered experimentally or therapeutically to the retina. PMID- 9418741 TI - Electroporation and bleomycin in glaucoma-filtering surgery. AB - PURPOSE: To develop a new treatment in glaucoma-filtering surgery using combined treatment with electroporation and the antiproliferative drug bleomycin. METHODS: Pigmented rabbits were treated with both bleomycin (10 micrograms/ml) and localized electric pulses (EP) using a special probe (5 V/cm, 100 msec, 8 pulses) (n = 10; group A). After EP treatment, bleomycin was washed out with 50 ml balanced salt solution, and then a posterior lip sclerectomy was performed on the same area. We also studied rabbits undergoing a posterior lip sclerectomy with bleomycin treatment alone (n = 10; group B), a posterior lip sclerectomy with EP treatment alone (n = 5; group C) and a posterior lip sclerectomy alone using the same operation (n = 5; group D, negative control). The intraocular pressure (IOP) was measured before and 1, 3, 5, 7, 10, 15, and 20 days after surgery. The formation of blebs, the conjunctiva, and the cornea were periodically examined by slit-lamp biomicrography. RESULTS: In every group, the IOP decreased until day 7, and no significant difference was observed among the four groups. In groups B, C, and D (control), the IOP increased gradually from day 10 and thereafter returned to the preoperative level after 15 days. However, in group A, the IOP remained lower than the preoperative level for 20 days; it was also significantly lower than each of the other three groups (P < 0.01). The survival rate of a filtering bleb was significantly higher in group A than in groups B, C, or D, but the survival rate in group B was not higher than groups C or D. No adverse effects were clinically observed in the ocular tissue, such as the cornea and conjunctiva. CONCLUSIONS: The combined treatment with EP and bleomycin was found to decrease IOP more prominently than EP or bleomycin treatment alone in filtering surgery. This treatment thus makes filtering surgery effective by decreasing the dose of the antiproliferative drug and by possibly localizing the drug delivery. PMID- 9418742 TI - Antidepressants in the treatment of premenstrual dysphoric disorder. AB - Premenstrual dysphoric disorder describes a subset of women who have severe premenstrual symptoms, including at least one mood symptom. It is included in DSM IV under "Depressive Disorders Not Otherwise Specified." Criteria differentiating premenstrual dysphoric disorder from premenstrual syndrome are the requirements that patients have at least five symptoms, including one mood symptom; have impairment associated with the illness; and prospectively confirm the symptoms. After decades of treatment research on premenstrual dysphoria, the most consistent positive results have been found for selective antidepressants, primarily those that are active at serotonin receptors. Most studies have used continuous daily treatment for acute phase therapy. Further studies should define the role of intermittent and long-term maintenance therapies with these agents. PMID- 9418743 TI - Antidepressants in the treatment of attention-deficit/hyperactivity disorder. AB - Antidepressants differ in their effectiveness for treating attention deficit/hyperactivity disorder (ADHD) in adults and children. None are as effective as psychostimulants for treating the attentional and cognitive symptoms, but they can help reduce impulsive and hyperactive behavior. Tricyclic antidepressants have well-demonstrated efficacy in treating behavioral symptoms, but desipramine should be avoided, at least in youths and adolescents (and perhaps adults), because safer tricyclics are available. Bupropion was effective in its few controlled trials, but tics and (especially in youth) skin rash limit its value. Venlafaxine appears effective, but controlled studies are needed. Serotonin selective reuptake inhibitors have not been tested in controlled trials, but they cause inconsistent changes, often aggravate ADHD symptoms, and can cause frontal apathy and disinhibition. Clonidine has not been adequately examined but seems to have small or uncertain effects. Psychostimulants remain the treatment of choice because of their unique effect on attention. Multimodal treatments (medications plus psychosocial) might not be more effective than medications alone. PMID- 9418744 TI - New uses for antidepressants: social phobia. AB - Data from recent epidemiologic surveys of community populations indicate that social phobia is a common psychiatric disorder and is associated with substantial functional impairment in a number of patients. Social phobia is also often comorbid with major depression, substance use disorders, and other anxiety disorders. Fortunately, a variety of antidepressant medications have been reported to alleviate the symptoms of social phobia. Controlled studies have shown substantial efficacy for the monoamine oxidase inhibitors phenelzine, moclobemide, and brofaromine and the serotonin selective reuptake inhibitors fluvoxamine and sertraline. Other serotonin reuptake inhibitors and venlafaxine have shown promise in case reports and open trials. PMID- 9418745 TI - New developments in the treatment of obsessive-compulsive disorder. AB - The treatment of obsessive-compulsive disorder (OCD) has changed dramatically in the last 10 years. Currently, the serotonin reuptake inhibitors (SRIs) and the serotonin selective reuptake inhibitors (SSRIs) are considered the "first choice" agents for pharmacologic treatment of OCD, although few head-to-head comparisons exist between any two specific agents. Strategies for nonresponders and partial responders to the SRI/SSRIs are reviewed. The only agents that have shown significant improvement as augmenting agents to an SRI/SSRI in systematic trials have been clonazepam and haloperidol. Predictors of response to pharmacotherapy have been limited, but several reports have found that an early age at onset of OCD has been associated with a poorer response to medications. Long-term maintenance medication may be necessary for some, although behavioral therapy may improve the need for extended pharmacotherapy. Cognitive behavioral therapy, specifically exposure with response prevention, still remains an effective and important component of treatment for many. One of the newest developments is the identification of a pediatric subtype of OCD characterized by prepubertal acute onset after group A beta-hemolytic streptococcal pharyngitis. Investigation trials with these children include immunomodulatory therapies and penicillin treatment and prophylaxis. If a unique subgroup of children with OCD can be identified, then novel treatments may prove effective and have a role in long term prophylaxis. PMID- 9418746 TI - Pharmacotherapy of borderline personality disorder. AB - Borderline personality disorder can be classified into four groups of symptoms: affective, impulsive, ego-interpersonal, and psychotic. Pharmacotherapy of borderline personality disorder should be directed at the severity of the symptoms in each of these groups, rather than by the presence or absence of the overall syndrome. This article reviews the pharmacotherapy of borderline personality disorder, with special emphasis on affective and impulsive symptoms. Overall, the MAOIs, the SSRIs, and the newer antidepressants (such as venlafaxine) provide the widest spectrum of effective treatment for the symptoms of borderline personality disorder. PMID- 9418748 TI - Turning data into management information. PMID- 9418747 TI - The use of newer antidepressants for panic disorder. AB - Antidepressants are frequently prescribed to treat panic disorder. Although tricyclic antidepressants and monoamine oxidase inhibitors both block panic attacks, they have many adverse effects such as orthostatic hypotension and weight gain. High potency benzodiazepines such as alprazolam are also efficacious but carry the risk of physical dependency. Data from research trials as well as clinical experience are accumulating to indicate that the serotonin selective reuptake inhibitors (SSRIs)--fluoxetine, fluvoxamine, paroxetine, and sertraline- and perhaps venlafaxine, which inhibits both serotonergic and noradrenergic reuptake, are useful antipanic medications. The possibility also exists that these newer antidepressants such as SSRIs and venlafaxine are superior in effectiveness to the previously available drugs and, when combined with cognitive behavioral therapy, might provide the best treatment outcome for patients with panic disorder. PMID- 9418749 TI - Proactive changes in nursing work force development in Tennessee. AB - Problems to be addressed by the CIC grant have been identified and defined, and a coalition has been formalized to address nursing work force development in Tennessee. The THCN, composed of consumers, nursing educators, policy makers, healthcare providers, and representatives of professional organizations and regulatory boards, is enthusiastically tackling the implementation phase of the project. These members are demonstrating open communication, teamwork, and interdependent decision making. Results are being achieved through persuasion, negotiation, seeking input from others, showing political sensitivity, and a willingness to share rewards and recognition. PMID- 9418750 TI - An administrative blunder. A nurse accused of mishandling controlled substances continues practicing. PMID- 9418751 TI - Nursing administration in Palestine. PMID- 9418752 TI - The nurse manager as career coach. AB - The conventional wisdom has been that career coaching is an important part of the nurse manager's role. In the current stressful healthcare environment, nurse managers seem more than ever in need of direction in this area. This paper examines the nurse manager's traditional role as a career coach, identifies the problems the current environment poses for implementation of that role and, considers options for achieving the goal of supporting staff with career development. PMID- 9418753 TI - Failure mode and effects analysis. An interdisciplinary way to analyze and reduce medication errors. AB - Medication errors are of interest both as an indication of quality of nursing care and as a measure of cost control. Developed for use in high-risk industries, Failure Mode and Effects Analysis used as an interdisciplinary approach in healthcare offers a comprehensive, individualized way to analyze and lessen medication errors in any institution. PMID- 9418754 TI - Staffing the operating room. Time and space factors. AB - Staffing the operating room requires a different approach than staffing inpatient units. Of course, there are similarities, namely: assuring safe, cost-effective care; dealing with fluctuating workloads; using various types of caregivers; and maximizing and maintaining resources in a responsible manner. However, many differences also exist. This article details the fundamental requirements for determining operating room staffing. PMID- 9418755 TI - Professional practice. A framework for transition to a new culture. AB - The reorganization of healthcare systems has largely focused on the redesign of clinical management structures with little emphasis on the systems that are essential to support professional practice and quality of care. This article describes the unique process of designing a framework that clearly articulates the essential elements of professional practice. The framework was crucial in clarifying account-abilities and role relationships between the clinical management and professional practice structures in the design and implementation of a major reorganization at Toronto East General Hospital, a 400-bed community teaching hospital in Toronto, Canada. PMID- 9418756 TI - The surgical hotline. Bridging the gap between hospital and home. AB - The telephone, an effective means of communication, offers the public access to healthcare personnel for advice and support. The authors describe the development of a surgical hotline, an innovative and cost-effective method that provides a continuum of care to postsurgical patients. PMID- 9418757 TI - Cutaneous vascular proliferation. Part II. Hyperplasias and benign neoplasms. AB - This second part of our review about vascular proliferations summarizes the clinicopathologic features of the cutaneous vascular hyperplasias and benign neoplasms. Hyperplasias comprise a heterogeneous group of vascular proliferations that eventually show a tendency to regression. Angiolymphoid hyperplasia with eosinophilia is included within the group of hyperplasias because of its historical denomination and its reactive nature, probably as a consequence of an arteriovenous shunt, although usually the lesions do not regress. Pyogenic granuloma, bacillary angiomatosis, intravascular papillary endothelial hyperplasia, and pseudo-Kaposi's sarcoma qualify as vascular hyperplasias because they regress when the stimulus that initiated them is removed. Benign neoplasms form a large group of hemangiomas with distinctive clinicopathologic characteristics, although some of them are of recent description and may produce diagnostic difficulties. We classified cutaneous benign vascular neoplasms according to their cell lineage of differentiation, for example, endothelial, glomus cell, and pericytic differentiation. Subsequent categories are established according to the size of the involved vessels (capillaries, venules and arterioles, or veins and arteries) or the nature of the proliferating vessels (blood or lymphatic vessels). Capillary and cavernous hemangiomas have been the terms classically used to name the most common variants of benign vascular neoplasms (i.e., infantile hemangiomas), but they are not the most appropriate denominations for these lesions. First, these names are not contrasting terms. Furthermore, most of the socalled "cavernous" hemangiomas are not hemangiomas (neoplasms) at all, but venous malformations. The most important conceptual issue is that, at any point in time, a particular hemangioma has its own histopathologic pattern throughout the depth of the lesion. For these reasons, we classified hemangiomas into superficial and deep categories. Some of the lesions reviewed have been recently described in the literature, and they may histopathologically mimic lesions of Kaposi's sarcoma; these include targetoid hemosiderotic hemangioma, microvenular hemangioma, tufted hemangioma, glomeruloid hemangioma, kaposiform hemangioendothelioma, spindle-cell hemangioendothelioma, and benign lymphangioendothelioma. In each of these lesions, we update and emphasize those clinical and histopathologic features that are helpful for differential diagnosis with lesions of authentic Kaposi's sarcoma in any of its three stages of development (patch, plaque, or nodule). PMID- 9418758 TI - Lichen amyloidosus: a consequence of scratching. AB - BACKGROUND: Lichen amyloidosus (LA) is generally said to be a pruritic type of amyloidosis of unknown cause. Histopathologically, it is characterized by epidermal changes of lichen simplex chronicus and by deposits of amyloid in the papillary dermis that are derived from keratin peptides of necrotic keratinocytes. Chronic scratching is responsible for the development of lichen simplex chronicus and may lead to necrosis of individual keratinocytes. OBJECTIVE: Our purpose was to evaluate whether chronic scratching may also be responsible for the formation of amyloid in LA. METHODS: We studied patients with LA in regard to histopathologic findings, onset of pruritus, associated diseases, and response to treatment. RESULTS: In most cases, pruritus had preceded the skin lesions. Eight of nine patients suffered from diseases other than LA that may be associated with pruritus. Histopathologically, amyloid was confined to areas that also showed signs of lichen simplex chronicus. Systemic treatment with sedating antihistamines and intense local treatment with corticosteroids were found to be effective. CONCLUSION: LA is considered to be a variant of lichen simplex chronicus in which scratching leads to necrosis of keratinocytes and eventually to the formation of amyloid in the papillary dermis. Because chronic scratching seems to be the cause and not the result of the deposits of amyloid, treatment should be directed at the amelioration of pruritus. PMID- 9418759 TI - The effects of topical calcipotriol on systemic calcium homeostasis in patients with chronic plaque psoriasis. AB - BACKGROUND: Calcipotriol is an effective treatment of chronic plaque psoriasis. We have previously demonstrated that it has a small effect on systemic calcium homeostasis even at recommended doses. OBJECTIVE: We attempted to determine the mechanism of the effect of calcipotriol on systemic calcium homeostasis so we could assess the possible consequences of long-term use. METHODS: Sixteen patients with extensive chronic plaque psoriasis were hospitalized and treated with high-dose topical calcipotriol. Up to 360 gm of calcipotriol (50 micrograms/gm) ointment was applied per week for 2 weeks under controlled conditions. RESULTS: There was a dose-dependent rise in intestinal absorption of calcium. No effect on bone turnover was demonstrated over this short period. Five patients became hypercalcemic, and there was a dose-dependent rise in serum total adjusted calcium, serum ionized calcium, serum phosphate, urine calcium, and urine phosphate. There was a dose-dependent fall in serum parathyroid hormone and serum 1,25 dihydroxyvitamin D3. CONCLUSION: Calcipotriol exerts its effects on systemic calcium homeostasis by increasing intestinal absorption of calcium and probably phosphate. This results in suppression of parathyroid hormone and 1,25 dihydroxyvitamin D3. PMID- 9418760 TI - Evaluation of the Ultraviolet Index: media reactions and public response. AB - BACKGROUND: In response to the increasing rate of skin cancer, particularly melanoma in the United States, the Environmental Protection Agency, the National Weather Service, the Centers for Disease Control and Prevention, National Association of Physicians for the Environment, and the American Academy of Dermatology, developed the Ultraviolet Index (UVI) to inform the public of the strength of the sun's rays and advise on methods for sun protection. OBJECTIVE: Our purpose was to evaluate the extent to which television stations and newspapers reported the UVI and assess the public's response to it. METHODS: To evaluate the effect of this effort, we surveyed television weather forecasters at 185 stations and examined weather pages in 54 newspapers in 58 cities that received the UVI reports. We also conducted a population probability telephone survey of 700 white adults (18 years of age and older) in these 58 cities. RESULTS: Seventy-one percent of the 169 stations that provided survey data for both 1994 and 1995 broadcast the UVI; 61% of newspapers reported the UVI. Nearly 64% of the 700 respondents (n = 445) had heard of the UVI. Of these respondents, 38% (n = 170) stated that they or their family changed their sun protection practices as a result of the UVI. CONCLUSION: The majority of television weather forecasters and newspapers reported the UVI. Most of the public was aware of the UVI, causing some to change sun protection practices. Further evaluation is required to maximize the effect of the UVI on sun protection practices. PMID- 9418761 TI - Normal vitamin D levels can be maintained despite rigorous photoprotection: six years' experience with xeroderma pigmentosum. AB - BACKGROUND: Although sun protection is advocated for skin cancer prevention, sunlight is also important in generation of vitamin D in the skin. There is concern that sun protection may result in an abnormally low level of vitamin D. OBJECTIVE: To assess the risk of vitamin D deficiency in a sunlight-deprived population, we studied eight ambulatory patients with xeroderma pigmentosum (XP) who practiced intensive sun protection during a chemoprevention study of oral isotretinoin. METHODS: We surveyed the patients to determine the extent of sun protection and vitamin D intake and measured the serum levels of two vitamin D metabolites (25-hydroxyvitamin D [25-OHD] and 1,25-dihydroxyvitamin D [1,25 (OH)2D]), calcium, and parathyroid hormone during 6 years. RESULTS: The patients all wore protective clothing and sunscreens when outdoors. Estimated mean vitamin D intake was normal. The mean values of serum 25-OHD were low normal, but 1,25 (OH)2D, calcium, ionized calcium and parathyroid hormone levels were normal. Lack of seasonal variation in serum 25-OHD indicated rigorous photoprotection. CONCLUSION: Despite rigorous sun protection normal vitamin D levels can be maintained in ambulatory patients with XP. PMID- 9418762 TI - Polarized light photography enhances visualization of inflammatory lesions of acne vulgaris. AB - BACKGROUND: Polarized light photography has been used to selectively differentiate surface from subsurface features of photoaged skin. OBJECTIVE: Our purpose was to compare acne assessments obtained from clinical evaluations with assessments from photographs obtained with flash photography and with perpendicular polarized light photography. METHODS: Assessments of acne with the Cunliffe scale were made of 32 subjects. Retrospective evaluations of standard and perpendicular polarized light photographs were made in a blinded fashion by a panel of evaluators. RESULTS: Visualization of inflammatory acne lesions was enhanced with perpendicular polarized light photography, with clear delineation of erythematous borders. Acne assessments with the use of a Cunliffe scale were significantly higher (p = 0.001) from perpendicular polarized light photographs than for clinical evaluations. CONCLUSION: Polarized light photography enhances visualization of inflammatory acne lesions in a manner not possible with conventional flash photographs, permitting accurate evaluation of the extent of disease and the effectiveness of therapy. PMID- 9418763 TI - Family physicians' knowledge of malignant melanoma. AB - BACKGROUND: The incidence and mortality of melanoma have been increasing during the past two decades. Melanoma is relatively easy to detect and, when diagnosed early, is curable. OBJECTIVE: Our purpose was to assess the baseline knowledge of malignant melanoma among family practitioners and to identify their preferred method of learning new information about the skin. METHODS: The design was a randomized survey by mailed questionnaire. RESULTS: More than 50% of physicians stated they lacked confidence in being able to recognize melanoma. Family physicians were well informed on factors affecting prognosis, but knowledge deficits were identified on history, physical examination, and risk factors. The most effective method of learning new information about the skin was hands-on teaching demonstration. CONCLUSION: Many family physicians are not confident in their ability to recognize melanoma, and knowledge deficits were identified. In addition, some important risk factors were not well recognized. Thus, those at highest risk may not be receiving education on early detection that may help decrease the incidence and mortality of melanoma. PMID- 9418764 TI - Optical coherence tomography of the human skin. AB - BACKGROUND: Optical coherence tomography (OCT) is a new diagnostic method for tissue characterization. OBJECTIVE: We investigated normal and pathologic structures in human skin in several locations to evaluate the potential application of this technique to dermatology. METHODS: Based on the principle of low-coherence interferometry, cross-sectional images of the human skin can be obtained in vivo with a high spatial resolution of about 15 microns. Within a penetration depth of 0.5 to 1.5 mm, structures of the stratum corneum, the living epidermis, and the papillary dermis can be distinguished. RESULTS: Different layers could be detected that were differentiated by induction of experimental blisters and by comparison with corresponding histologic sections. Furthermore, OCT images of several skin diseases and tumors were obtained. CONCLUSION: OCT is a promising new imaging method for visualization of morphologic changes of superficial layers of the human skin. It may be useful for noninvasive diagnosis of bullous skin diseases, skin tumors, and in vivo investigation of pharmacologic effects. PMID- 9418765 TI - Patch testing in children and adolescents: five years' experience and follow-up. AB - BACKGROUND: Allergic contact dermatitis in children is a significant clinical problem. Little information is available concerning the value of patch testing and the outcome in these children. OBJECTIVE: Our purpose was to assess the value of patch testing in children and the outcome of allergic contact dermatitis in childhood. METHODS: Clinical data on 83 children patch tested during a 5-year period were assessed. Clinical follow-up on 68 subjects was performed. RESULTS: Overall, 41 children had one or more allergic reactions (49%). Reactions to metals, topical preparations, and food additives were common. The clinical outcome at 6 months was significantly better for 36 children with a relevant allergen on patch testing than in 32 with no allergen or no relevant allergen (p = 0.006). CONCLUSION: Patch testing is useful in the management of children suspected of having an allergic contact dermatitis. Patch testing and subsequent allergen avoidance may improve the prognosis in children with a relevant contact allergen. PMID- 9418766 TI - Itraconazole pulse therapy for onychomycosis and dermatomycoses: an overview. AB - BACKGROUND: Itraconazole is a broad-spectrum antifungal agent that has been used to treat dermatomycosis and onychomycosis using continuous therapy. More recently the drug has been used as pulse dosing. OBJECTIVE: Our purpose was to review the studies in which itraconazole pulse therapy (PT) has been administered in the management of dermatomycoses. RESULTS: For tinea pedis and manuum, the recommended dosage is itraconazole 200 mg twice daily for 1 week (n = 220). A clinical response and mycologic cure rate of 90% +/- 4% and 76% +/- 6%, respectively, has been obtained. For tinea corporis/cruris, itraconazole 200 mg/day for 1 week (n = 354) resulted in a clinical response and mycologic cure rate of 90% +/- 4% and 77% +/- 6%, respectively. When three pulses of itraconazole are used to treat toenail onychomycosis (n = 1389), the clinical cure rate, clinical response, and mycologic cure rate at follow-up 12 months after the start of therapy were 58% +/- 10%, 82% +/- 3%, and 77% +/- 5%, respectively. With two pulses for onychomycosis of the fingernails, the clinical cure rate, clinical response, and mycologic cure rate at follow-up, 9 months after the start of therapy, were 78% +/- 10%, 89% +/- 6%, and 87% +/- 8%, respectively. CONCLUSION: Itraconazole PT is effective and safe in the treatment of tinea pedis/manuum, tinea corporis/cruris, and onychomycosis. PMID- 9418767 TI - The staged retroauricular to auricular direct pedicle (interpolation) flap for helical ear reconstruction. AB - BACKGROUND: A significant soft-tissue defect involving the helix of the external ear may present a difficult challenge to repair. OBJECTIVE: We describe our experience with the staged retroauricular to auricular pedicle flap for repair of soft-tissue defects with exposed cartilage of the helix of the ear. METHODS: The staged pedicle flap was used to repair 26 helical ear defects after excision of basal cell carcinoma (n = 16), squamous cell carcinoma (n = 3), and melanoma (n = 7). RESULTS: Defect size ranged from 1 x 2 cm to 4 x 6 cm (average 2.4 x 3.3 cm). Defects involved the superior helix in 12 patients, mid helix in 11, and inferior helix in three. No cases of infection or flap necrosis occurred. CONCLUSION: The staged retroauricular to auricular pedicle flap consistently provides a good to excellent functional and cosmetic outcome when performed on properly selected helical ear defects. PMID- 9418769 TI - Practical clinical dermatology: photography with professional results. AB - During the last decade our office use of photographs to document patient treatment has increased. Many photographs were wasted because of poor quality, others because of mismatched views. Storage was cumbersome, and retrieval was time-consuming. The objective was to produce better quality photographs cost effectively, with a user-friendly camera system. A professional consultant solved these problems. A specific camera and system to document and take photographs was designed that can be easily implemented in any office practice. PMID- 9418768 TI - Terbinafine: an update. AB - Oral terbinafine was first introduced in the United Kingdom in February 1991 and was approved for the treatment of onychomycosis in the United States in May 1996. It is estimated that 4 million patients worldwide have been treated with oral terbinafine as of December 1996. The efficacy of terbinafine in the treatment of onychomycosis and other dermatomycoses is reviewed. The adverse-effects profile of oral terbinafine is evaluated. PMID- 9418770 TI - Surgical pearl: clavicular grafts are "superior" to supraclavicular grafts. PMID- 9418771 TI - Purpuric drug eruption secondary to itraconazole. PMID- 9418773 TI - Premature familial sebaceous hyperplasia: successful response to oral isotretinoin in three patients. PMID- 9418772 TI - Interpretation of potassium hydroxide wet mounts by means of telemicroscopy. PMID- 9418774 TI - Notalgia paresthetica associated with nerve root impingement. PMID- 9418775 TI - Skin cancer associated with ichthyosis: the MAUIE syndrome. PMID- 9418776 TI - Langerhans cell histiocytosis mimicking child abuse. PMID- 9418777 TI - Linear chronic cutaneous graft-versus-host disease. PMID- 9418778 TI - Painful skin papules caused by concomitant Acremonium and Fusarium infection in a neutropenic child. PMID- 9418779 TI - Cutaneous squamous cell carcinoma with zosteriform metastasis in a transplant recipient. PMID- 9418780 TI - Keratoacanthoma centrifugum marginatum: treatment with intralesional bleomycin. PMID- 9418781 TI - Primary cutaneous B-cell lymphoma: a clinically homogeneous entity? PMID- 9418782 TI - Polysensitivity in fixed drug eruption. PMID- 9418783 TI - Periungual erythema in HIV-infected patients. PMID- 9418784 TI - Photodermatology. PMID- 9418785 TI - Multimodal management of diffuse neonatal hemangiomatosis. PMID- 9418786 TI - Chronic telogen effluvium: potential complication for clinical trials in female androgenetic alopecia? PMID- 9418787 TI - Cutaneous localization of B-cell chronic lymphocytic leukemia at the site of varicella/herpesvirus eruptions. PMID- 9418788 TI - Special pleading at Kyoto. PMID- 9418789 TI - Cloning: potential benefits for human medicine. PMID- 9418791 TI - Medical information and the World Wide Web. PMID- 9418790 TI - Medicine, modernism and after: a new role model for the doctor? PMID- 9418792 TI - Information technology in medicine. PMID- 9418793 TI - Facsimile communication between emergency departments and GPs, and patient data confidentiality. AB - OBJECTIVE: To assess general practitioners' perceptions of the effectiveness of facsimile notification of their patients being admitted from the emergency department (ED), and its adequacy in terms of patient confidentiality. DESIGN: Questionnaire survey, before and after the initiation of facsimile notification. SETTING: A provincial community of approximately 120,000 residents in Victoria. MAIN OUTCOME MEASURES: Changes in GPs' ratings of communication with the ED; acceptability of facsimile notification; and concerns about patient confidentiality. RESULTS: 77 of 85 GPs participated; only 44 (57.1%) returned both questionnaires. ED-GP communication ratings of "adequate" or better increased from 48% to 100% (P < 0.05). The proportion of GPs who were notified of all admissions increased from 0 to 41% (P < 0.05). The proportion of GPs who preferred facsimile for notification increased from 39% to 68% (P < 0.05). Most GPs found the initiative acceptable and reservations about confidentiality decreased from 36% to 16% (P < 0.05). 38 of the 887 patients admitted from the ED (4.3%) refused facsimile notification. CONCLUSIONS: Facsimile improves ED-GP communications and may, in turn, improve the quality and continuity of patient care. Informed consent should be obtained from all patients. PMID- 9418794 TI - Usefulness of a patient library in a suburban general practice. AB - OBJECTIVES: To provide health information by means of a patient library in an Australian general practice, and to examine the effects of such information on patient knowledge, anxiety and behaviour. DESIGN: Questionnaire survey of patients who borrowed library items and of doctors within the practice. SETTING: A five-doctor suburban practice serving approximately 7000 patients. RESULTS: 210 items were purchased and made available for loan over the two years of the study; 164 questionnaires were obtained after 229 borrowings in the first 15 months. Patients who borrowed items were mostly women (88%) and a high proportion of borrowers (47%) felt that the doctors' surgery was their main source of health information. 98% felt that borrowing an item had been "useful" or "very useful", 88% felt that it had increased their knowledge, 48% felt it had decreased their anxiety, and 79% felt it had changed their behaviour. The doctors were generally positive about the library. CONCLUSIONS: The practice library was a very useful addition to the surgery services and provided a valuable clinical tool for the doctors. PMID- 9418795 TI - Telemedicine ophthalmology consultation in remote Queensland. AB - OBJECTIVE: To assess the use of remote telemedicine ophthalmology in patients presenting to an emergency department with acute eye problems. DESIGN: A prospective review from 1 December 1996 to 28 February 1997 of referral patterns and telemedicine consultations, comparing referral patterns with the same period one year before. PARTICIPANTS AND SETTING: 24 patients presenting to the emergency department of a remote base hospital in Queensland with an acute ophthalmological problem requiring a specialist opinion. MAIN OUTCOME MEASURES: Clinical outcomes; use of the Patient Transit Scheme for isolated patients; acceptability to patients and doctors; and ophthalmologists' opinions of the system. RESULTS: No adverse outcomes were identified. Patients transferred for urgent assessment fell from 17 for the corresponding period in the previous year to four during the study period, while respective numbers of patients requiring non-urgent transfers (for surgery or postoperative review) during the same periods were 41 and 30. Both patients and staff (including the ophthalmologists) found the telemedicine facility very acceptable. CONCLUSION: Ophthalmology is well suited to telemedicine for the diagnosis and management of acute conditions and postoperative assessment of patients in remote areas. It offers considerable potential benefits to patients, and enhances the skills of local practitioners. PMID- 9418796 TI - Forty years of plotting for public health. PMID- 9418797 TI - MaLAM, a medical lobby for appropriate marketing of pharmaceuticals. PMID- 9418798 TI - Medical planning for the Sydney 2000 Olympic and Paralympic Games. PMID- 9418799 TI - Medical and public health services at the 1996 Atlanta Olympic Games: an overview. AB - Planning for the 2000 Sydney Olympic Games may benefit from the experience of the 1996 Atlanta Olympics. Excellent health promotion and prevention activities before and during the Games resulted in fewer medical and public health problems than anticipated. Despite this, there was room for improvement in the level of communication and cooperation between the many service providers to ensure the most appropriate and efficient responses. PMID- 9418800 TI - The Polyclinic at the 1996 Atlanta Olympic Village. AB - The Polyclinic, staffed mainly by volunteers, successfully provided primary health care during 16,519 patient encounters, 64% involving athletes. However, the profile of patient needs held some surprises. PMID- 9418801 TI - Hospital use by Olympic athletes during the 1996 Atlanta Olympic Games. PMID- 9418802 TI - Funding Australia's health and medical research. PMID- 9418803 TI - Mapping Australia's basic research in the medical and health sciences. AB - The Institute for Scientific Information indexes most of the major international basic research journals in science in the Science Citation Index (SCI). Australia's presence in the medical and health sciences journals in the SCI and the citations its published research receives in these journals show that Australia's basic medical research has high international "visibility". Mapping the source of the most highly "visible" Australian medical research articles shows high impact research coming from several different sectors (research institutes, universities, hospitals, etc.), but with a concentration in the member institutions of the Australian Association of Medical Research Institutes (AAMRI). Published research from the AAMRI is cited at a rate two-thirds higher than the Australian average for medical and health sciences. PMID- 9418804 TI - The Institute of Medical and Veterinary Science. PMID- 9418805 TI - Doctoring beyond frontiers. PMID- 9418806 TI - Safe motherhood: impossible dream or achievable reality? PMID- 9418808 TI - The crossing. PMID- 9418807 TI - Medical log: forward command Thredbo. PMID- 9418809 TI - On the water's edge. PMID- 9418810 TI - Crisis management in the community. PMID- 9418811 TI - From shunned to shining: doctors, madness and psychiatry in Australian and New Zealand cinema. PMID- 9418812 TI - The famous five F's: focus on fluid. PMID- 9418814 TI - Faith healing or Russian roulette? PMID- 9418813 TI - Wicket-keeper bowled (over) by (eye) ball. Uveal melanoma with spontaneous hyphaema. PMID- 9418815 TI - "Irukandji" syndrome: a risk for divers in tropical waters. PMID- 9418816 TI - Papilloedema and coma in a child: undescribed symptoms of the "Irukandji" syndrome. PMID- 9418817 TI - European wasps: an emerging hazard in Australia. PMID- 9418818 TI - Choking after inhaling a foreign body through a Ventolin puffer. PMID- 9418819 TI - Out of the blue and into the pink. A new litmus test for chlorine gas exposure. PMID- 9418820 TI - "Hale-Bopp" and "Knocking on Heaven's Gate". Hits of the Net, 1997. PMID- 9418821 TI - Rotavirus gastroenteritis: is vaccine prevention near at hand? PMID- 9418822 TI - A longitudinal study of the attitudes of the medical profession towards competition and advertising. AB - AIMS: To measure changes in the attitudes of medical practitioners toward the move from a collegial to a more competitive orientation of the medical profession and, in particular, toward the role of advertising. METHOD: In the years 1985, 1988 and 1994, self-completion questionnaires were mailed to samples of medical practitioners. The questionnaires for each year were identical, containing forty Likert scales with questions relating to advertising, competition and commercial behaviour. Similar data were also gathered from members of the dental, veterinary, legal and accounting professions. RESULTS: Over the three studies there has been an undramatic but steady movement towards acceptance of a competitive orientation in general and towards acceptance of informative advertising in particular. There remains substantial reservation towards the use of persuasive advertising. Although patients are viewed as being more demanding than in past times, practitioners still expect their relationships with patients to be long lasting. CONCLUSION: The results suggest that acceptance of the move towards a more competitively oriented profession will continue. The results also suggest that in view of the kinds of advertising that are now used by medical practices, and of the change in attitudes that has occurred, control of advertising might cease to be a concern to the profession. PMID- 9418823 TI - A general practice case-control study of delayed immunisation in under two year old children. AB - AIM: To identify social factors which characterise the household of children with delayed immunisation. METHOD: The study was done in 15 general practices in the Wellington city region with a case-control design where preschool children who were not up to date (cases) for their immunisations were compared with children who were up to date (controls). RESULTS: There were 215 cases where immunisation was delayed among 3723 children at the time of audit in June 1996. Ethnic status was available from general practice records in 33% of cases and 40% of controls. There were more Maori and Pacific Island children among the cases compared to controls (39% versus 21%, odds ratio (OR) = 2.39, 95% confidence interval (CI) 1.12-5.10, p = 0.022. CASE SAMPLE HOUSEHOLDS COMPARED WITH CONTROLS HAD: (1) more children in the households (mean 2.27 versus 1.98, p = 0.01); (2) more households with a female as the only adult (35% versus 24%, OR = 1.61, 95% CI 1.04-2.51, p = 0.034). There was no difference in the proportions of male only adults (0.02% versus 0.01%, p = 0.70); (3) more mothers who were under 30 years of age (38% versus 25%, OR = 1.61, 95% CI 1.04-2.51, p = 0.034); and more fathers under 30 years of age (20% versus 10%, OR = 2.16, 95% CI 0.97-4.84, p = 0.06); (4) fewer mothers who were up to date with their recorded cervical smear status (66% versus 83%, OR = 0.69, 95% CI 0.23-0.63, p < 0.001); (5) more parents with an active community services card (38% versus 25%, OR 1.82, 95% CI 1.17-2.82, p = 0.007); (6) fewer siblings being immunised (70% versus 94%, OR = 0.14, 95% CI 0.07-0.28, p < 0.001). CONCLUSION: There is a distinct group of New Zealand children for whom immunisation is delayed. Such children live in households which can be characterised by various social factors easily obtainable from general practice records. The identified households could be targeted for efficient preventive care by general practices. PMID- 9418824 TI - Perceptions of diabetes among rural Maori elders and spokespersons. AB - AIMS: To assess knowledge and opinions of diabetes among rural Maori elders and spokespersons. METHODS: Interviews were conducted in rural South Auckland. Subjects were identified through their affiliation with one marae (meeting house), residence near the marae and being recognised locally as either male (kaumatua) or female (kuia) elders or spokespersons. The main researcher was a kuia chosen by and from within the local community. Interviews were conducted with 43/44 (98%) subjects identified. RESULTS: While specific diabetes knowledge was low, diabetes was seen, along with cancer, as one of the two major health issues for Maori. Results need to be understood in the context of the holistic understanding of health by Maori. CONCLUSION: The recognition of diabetes as a major health problem was accompanied by a call for diabetes education in a form that will generate interest and participation by Maori. It is timely for the introduction of marae based diabetes awareness and sustained exercise programmes as part of a diabetes prevention and control strategy among Maori communities where diabetes risk is high. PMID- 9418825 TI - Lead based paint hazards in early childhood centres in the Wellington region. AB - AIM: To determine the frequency of exposure to lead based paint (LBP) in early childhood centres (ECCs) in the Wellington region. METHODS: Senior staff at sixty nine randomly selected licensed ECCs in the Wellington region were contacted. Fifty two centres consented to participate. Senior staff at these centres were interviewed and centres built before 1980 were examined for damaged LBP surfaces accessible to children. The survey was carried out between July 1995 and January 1996. A centre was considered to be LBP positive if it had at least one damaged LBP surface accessible to children in the course of a normal day's activities. RESULTS: Fifty two per cent of surveyed centres were LBP positive. LBP hazards were detected in 60% of buildings built before 1965 and 30% of buildings constructed between 1965 and 1980, and were found more frequently in Te Kohanga Reo and childcare centres. Although children aged 2 years and under constituted only 27% of all children attending surveyed centres, over half (53%) of the children in this age group attended centres with an LBP hazard. Staff at ECCs were often uncertain about the causes and outcomes of lead toxicity and expressed a desire for further education. CONCLUSION: There is significant potential for exposure to lead in early childhood centres in Wellington as a consequence of accessible damaged lead based paint surfaces. PMID- 9418826 TI - Potent topical steroid in a Chinese herbal cream. AB - Steroids have been discovered previously in oral "herbal" preparations. Using liquid chromatography, we have now confirmed the presence of the potent topical steroid, clobetasol proprionate, in a Chinese herbal cream. PMID- 9418827 TI - Management guidelines for progressive chronic renal failure. New Zealand Nephrologists Consensus Group. AB - Consensus guidelines are provided for the conservative management of adult patients with chronic and progressive renal failure, together with a brief review of the evidence relating to various treatable complications. Blood pressure control, diet, hyperlipidaemia, calcium and phosphate metabolism, anaemia and acidosis are considered. PMID- 9418828 TI - Colorectal cancer screening. PMID- 9418829 TI - Colorectal cancer screening. PMID- 9418830 TI - Colorectal cancer screening. PMID- 9418831 TI - Rationing New Zealand's publicly funded health resources. PMID- 9418832 TI - Good news for people with spinal cord injuries. PMID- 9418833 TI - Expert system technology: a new aid for the gynaecologist in managing stress urinary incontinence. PMID- 9418834 TI - Disclosing a diagnosis of cancer. PMID- 9418835 TI - Hormone replacement therapy, diabetes and pancreatitis secondary to hypertriglyceridaemia. PMID- 9418836 TI - Controlling dangerous dogs. PMID- 9418837 TI - Molecular analysis of HLA-H gene mutations in New Zealand patients with haemochromatosis. AB - AIM: To determine the frequency of HLA-H gene mutations in New Zealand patients with haemochromatosis. METHODS: The Cys282Tyr and His63Asp mutations in the HLA-H gene were analyzed by polymerase chain reaction, restriction enzyme digestion and electrophoresis in two separate patient groups. The first was a group of 20 Christchurch patients with a definite clinical diagnosis of haemochromatosis. The second group consisted of 33 patients, with a provisional diagnosis of haemochromatosis, attending Dunedin Hospital for therapeutic venesection. RESULTS: All 20 Christchurch patients and 25 of the 33 (76%) Dunedin patients were homozygous for the Cys282Tyr mutation. After review of the clinical data, histology and response to venesection a diagnosis of haemochromatosis could be confidently excluded in six of the remaining eight patients. Despite atypical features, a diagnosis of haemochromatosis could not be excluded in the final two patients, one of whom was a compound heterozygote for the two mutations. CONCLUSIONS: Homozygosity for the Cys282Tyr mutation is closely associated with haemochromatosis in New Zealand patients. Molecular analysis of the HLA-H gene is indicated in the assessment of patients with iron overload including those currently being treated by venesection. PMID- 9418838 TI - Follow up of elderly patients after cardiac surgery and intensive care unit admission, 1991 to 1995. AB - AIMS: To examine the outcome of cardiac surgery and resulting intensive care admission in elderly (> or = 75 years) cardiac surgery patients at Waikato Hospital, 1991 to 1995. METHODS: Clinical records of all elderly cardiac surgery patients admitted to the intensive care unit were reviewed. All survivors were sent a postal questionnaire evaluating cardiac related symptom control and quality of life (QOL). Outcomes in the 'old' (75-79 years) and in the 'very old' (> or = 80 years) were compared. RESULTS: Seventeen of 97 patients had died. Mean survival time was 32.2 months. Survivor followup (100%) was at a mean of 34.8 months. Mean functional class (New York Heart Association or Canadian Cardiovascular Society) improved from 3.0 preoperatively to be 1.7. Cardioactive medications fell by a mean of 0.7 drugs per patient. Twenty-seven percent of survivors became more dependent as assessed by domicile type. Outcomes between the two groups were not different except for some improved individual changes in functional class. The 'very old' group have a similar postoperative QOL to that of the 'old' group. Ninety-two percent of survivors indicated that they would opt for cardiac surgery again if given the time over. CONCLUSIONS: Following cardiac surgery and intensive care admission at Waikato Hospital, surviving elderly patients have experienced a favourable outcome in terms of symptom control and quality of life. Mortality rates are acceptably low. PMID- 9418840 TI - The effect of frequent vacuum cleaning on the house dust mite allergen, Der p 1 in carpets: a pilot study. AB - AIM: To determine the effect of frequent vacuum cleaning of carpets on Der p 1, the major group one allergen of the house dust mite Dermatophagoides pteronyssinus. METHODS: Nine rooms and three hallways in the resident medical officers quarters at Wellington Hospital were regularly vacuum cleaned (daily, except weekends) over a five week period. Dust samples were collected before and at weekly intervals for Der p 1 measurement by a double monoclonal antibody ELISA technique. RESULTS: Der p 1 concentrations progressively declined from an initial geometric mean level (95% CI) of 4.47 micrograms/g fine dust (0.11-21.73) to 1.83 micrograms/g (0.29-9.57) after five weeks, a mean reduction of 48.0% (31.7-65.5). Similarly when expressed per unit area Der p 1 levels declined from 6.35 micrograms/m2 (1.15-35.16) to 1.66 micrograms/m2 (0.33-9.04), a mean reduction of 68.5% (58.6-78.3). CONCLUSIONS: Frequent vacuum cleaning over a short time significantly reduces house dust mite allergen levels in carpets. Larger long term trials are warranted to determine if greater reductions are possible that would be beneficial to house dust mite sensitive individuals. PMID- 9418839 TI - Lispro insulin as premeal therapy in type 1 diabetes: comparison with Humulin R. AB - AIMS: To determine the efficacy, tolerability and safety of the short-acting insulin analogue lispro compared with regular short-acting insulin, Humulin R as premeal therapy in type 1 diabetes mellitus and to assess the safety of lispro administered for one year. METHODS: The study was part of an international multicentre crossover study (IOAG) in which 1008 patients were randomised. Twenty patients from Auckland, with insulin dependent diabetes mellitus, received lispro for 3 months and Humulin R for 3 months in a crossover design. At the end of the crossover period, 19 patients elected to participate in an open label continuation of lispro therapy. Humulin N, L or U was used as basal insulin therapy. RESULTS: Lispro and Humulin R in combination with Humulin N, L or U did not significantly differ with respect to glycaemic control or incidence of hypoglycaemia. Glycosylated haemoglobin (HbA1C) improved from 8.6% at baseline to 7.6 +/- 0.9 (Humulin R) and 7.7 +/- 1.1% (lispro). During the open label continuation of lispro plus the usual basal insulin HbA1C deteriorated to 8.6% after 12 months. CONCLUSIONS: In this short-term comparison, lispro and Humulin R were well tolerated, while glycaemic control, incidence of hypoglycaemia and adverse effects were similar. PMID- 9418841 TI - Immunisation coverage in Christchurch in a birth cohort. Christchurch Immunisation Coordination Committee. AB - AIM: To ascertain the immunisation status of a birth cohort of infants at 8 months of age. METHODS: Data on all children born in Christchurch in June, July and August 1995 were matched with immunisation benefit claim information for the 6-week, 3-month and 5-month immunisation events. Those not listed as receiving all three immunisations by the age of 8 months were traced to ascertain their immunisation status. RESULTS: Immunisation status was established for 933 (93.1%) of the cohort of 1002 infants. Of these, 95% had received all three immunisations that were scheduled up to 5 months of age. This represents a coverage of at least 88.2% of the entire cohort. It was probable that many of those not traced would also have been fully immunised. Using a conservative estimate of coverage among this group, we calculated that 93% of the entire cohort had been fully immunised by the age of 8 months. CONCLUSION: There was a high immunisation coverage rate in Christchurch. This is in contrast to a commonly held view that children in New Zealand have a poor immunisation record. There was no evidence of a significant anti-immunisation stance among Christchurch parents. Immunisation data relevant to specific, defined populations can identify individual children and groups of children who may require assistance beyond general practitioner recall to achieve appropriate immunisation. The tracking process also allowed for immunisation reminders to be personally delivered and included some facilitation for achieving further immunisation. We suggest that a pilot programme, using a centralised register based on birth data, feeding into existing immunisation delivery and recall systems, and using tracking methods similar to those used in our study, should be the next step in addressing immunisation coverage surveillance and immunisation coordination in Christchurch. PMID- 9418842 TI - Hypersensitivity to seminal fluid. PMID- 9418843 TI - Monitoring progress towards targets for the prevalence of smoking in pregnancy. PMID- 9418844 TI - Prescribing data and cost comparisons in New Zealand. PMID- 9418845 TI - Gatekeepers no more. PMID- 9418846 TI - Relevance in health research funding: why it is important. PMID- 9418848 TI - Involvement of negative cofactor NC2 in active repression by zinc finger homeodomain transcription factor AREB6. AB - The transcription factor AREB6 contains a homeodomain flanked by two clusters of Kruppel type C2H2 zinc fingers. AREB6 binds to the E-box consensus sequence, CACCTGT, through either the N- or the C-terminal zinc finger cluster. To gain insights into the molecular mechanism by which AREB6 activates and represses gene expression, we analyzed the domain structure of AREB6 in the context of a heterologous DNA-binding domain by transient-transfection assays. The C-terminal region spanning amino acids 1011 to 1124 was identified as a conventional acidic activation domain. The region containing amino acids 754 to 901, which was identified as a repression domain, consists of 40% hydrophobic amino acids displaying no sequence similarities to other known repression domains. This region repressed transcription in vitro in a HeLa nuclear extract but not in reconstituted transcription systems consisting of transcription factor IID (TFIID), TFIIB, TFIIE, TFIIH/F, and RNA polymerase II. The addition of recombinant negative cofactor NC2 (NC2alpha/DRAP1 and NC2beta/Dr1) to the reconstituted transcription system restored the activity of the AREB6 repression domain. We further demonstrated interactions between the AREB6 repression domain and NC2alpha in yeast two-hybrid assay. Our findings suggest a mechanism of transcriptional repression that is mediated by the general cofactor NC2. PMID- 9418847 TI - Tau91, an essential subunit of yeast transcription factor IIIC, cooperates with tau138 in DNA binding. AB - Transcription factor IIIC (TFIIIC) (or tau) is a large multisubunit and multifunctional factor required for transcription of all class III genes in Saccharomyces cerevisiae. It is responsible for promoter recognition and TFIIIB assembly. We report here the cloning and characterization of TFC6, an essential gene encoding the 91-kDa polypeptide, tau91, present in affinity-purified TFIIIC. Tau91 has a predicted molecular mass of 74 kDa. It harbors a central cluster of His and Cys residues and has basic and acidic amino acid regions, but it shows no specific similarity to known proteins or predicted open reading frames. The TFIIIC subunit status of tau91 was established by the following biochemical and genetic evidence. Antibodies to tau91 bound TFIIIC-DNA complexes in gel shift assays; in vivo, a B block-deficient U6 RNA gene (SNR6) harboring GAL4 binding sites was reactivated by fusing the GAL4 DNA binding domain to tau91; and a point mutation in TFC6 (tau91-E330K) was found to suppress the thermosensitive phenotype of a tfc3-G349E mutant affected in the B block binding subunit (tau138). The suppressor mutation alleviated the DNA binding and transcription defects of mutant TFIIIC in vitro. These results indicated that tau91 cooperates with tau138 for DNA binding. Recombinant tau91 by itself did not interact with a tRNA gene, although it showed a strong affinity for single-stranded DNA. PMID- 9418849 TI - Novel IkappaB alpha proteolytic pathway in WEHI231 immature B cells. AB - The Rel/NF-kappaB family of transcription factors is sequestered in the cytoplasm of most mammalian cells by inhibitor proteins belonging to the IkappaB family. Degradation of IkappaB by a phosphorylation-dependent ubiquitin-proteasome (inducible) pathway is believed to allow nuclear transport of active Rel/NF kappaB dimers. Rel/NF-kappaB (a p50-c-Rel dimer) is constitutively nuclear in murine B cells, such as WEHI231 cells. In these cells, p50, c-Rel, and IkappaB alpha are synthesized at high levels but only IkappaB alpha is rapidly degraded. We have examined the mechanism of IkappaB alpha degradation and its relation to constitutive p50-c-Rel activation. We demonstrate that all IkappaB alpha is found complexed with c-Rel protein in the cytoplasm. Additionally, rapid IkappaB alpha proteolysis is independent of but coexistent with the inducible pathway and can be inhibited by calcium chelators and some calpain inhibitors. Conditions that prevent degradation of IkappaB alpha also inhibit nuclear p50-c-Rel activity. Furthermore, the half-life of nuclear c-Rel is much shorter than that of the cytoplasmic form, underscoring the necessity for its continuous nuclear transport to maintain constitutive p50-c-Rel activity. We observed that IkappaB beta, another NF-kappaB inhibitor, is also complexed with c-Rel but slowly degraded by a proteasome-dependent process in WEHI231 cells. In addition, IkappaB beta is basally phosphorylated and cytoplasmic. We thus suggest that calcium-dependent IkappaB alpha proteolysis maintains nuclear transport of a p50-c-Rel heterodimer which in turn activates the synthesis of IkappaB alpha, p50, and c-Rel to sustain this dynamic process in WEHI231 B cells. PMID- 9418850 TI - Proteasome inhibitors cause induction of heat shock proteins and trehalose, which together confer thermotolerance in Saccharomyces cerevisiae. AB - An accumulation in cells of unfolded proteins is believed to be the common signal triggering the induction of heat shock proteins (hsps). Accordingly, in Saccharomyces cerevisiae, inhibition of protein breakdown at 30 degrees C with the proteasome inhibitor MG132 caused a coordinate induction of many heat shock proteins within 1 to 2 h. Concomitantly, MG132, at concentrations that had little or no effect on growth rate, caused a dramatic increase in the cells' resistance to very high temperature. The magnitude of this effect depended on the extent and duration of the inhibition of proteolysis. A similar induction of hsps and thermotolerance was seen with another proteasome inhibitor, clasto-lactacystin beta-lactone, but not with an inhibitor of vacuolar proteases. Surprisingly, when the reversible inhibitor MG132 was removed, thermotolerance decreased rapidly, while synthesis of hsps continued to increase. In addition, exposure to MG132 and 37 degrees C together had synergistic effects in promoting thermotolerance but did not increase hsp expression beyond that seen with either stimulus alone. Although thermotolerance did not correlate with hsp content, another thermoprotectant trehalose accumulated upon exposure of cells to MG132, and the cellular content of this disaccharide, unlike that of hsps, quickly decreased upon removal of MG132. Also, MG132 and 37 degrees C had additive effects in causing trehalose accumulation. Thus, the resistance to heat induced by proteasome inhibitors is not just due to induction of hsps but also requires a short-lived metabolite, probably trehalose, which accumulates when proteolysis is reduced. PMID- 9418851 TI - Lagging-strand, early-labelling, and two-dimensional gel assays suggest multiple potential initiation sites in the Chinese hamster dihydrofolate reductase origin. AB - There is general agreement that DNA synthesis in the single-copy and amplified dihydrofolate reductase (DHFR) loci of CHO cells initiates somewhere within the 55-kb spacer region between the DHFR and 2BE2121 genes. However, results of lagging-strand, early-labelling fragment hybridization (ELFH), and PCR-based nascent-strand abundance assays have been interpreted to suggest a very narrow zone of initiation centered at a single locus known as ori-beta, while two dimensional (2-D) gel analyses suggest that initiation can occur at any of a large number of potential sites scattered throughout the intergenic region. The results of a leading-strand assay and two intrinsic labelling techniques are compatible with a broad initiation zone in which ori-beta and a second locus (ori gamma) are somewhat preferred. To determine how these differing views are shaped by differences in experimental manipulations unrelated to the biology itself, we have applied the lagging-strand, ELFH, neutral-neutral, and/or neutral-alkaline 2 D gel assays to CHOC 400 cell populations synchronized and manipulated in the same way. In our experiments, the lagging-strand assay failed to identify a template strand switch at ori-beta; rather, we observed a gradual, undulating change in hybridization bias throughout the intergenic spacer, with hybridization to the two templates being approximately equal near a centered matrix attachment region. In the ELFH assay, all of the fragments in the 55-kb intergenic region were labelled in the first few minutes of the S phase, with the regions encompassing ori-beta and ori-gamma being somewhat preferred. Under the same conditions, neutral-neutral and neutral-alkaline 2-D gel analyses detected initiation sites at multiple locations in the intergenic spacer. Thus, the results of all existing replicon-mapping methods that have been applied to the amplified DHFR locus in CHOC 400 cells are consistent with a model in which two somewhat preferred subzones reside in a larger zone of multiple potential initiation sites in the intergenic region. PMID- 9418852 TI - RNA recognition motif 2 of yeast Pab1p is required for its functional interaction with eukaryotic translation initiation factor 4G. AB - The eukaryotic mRNA 3' poly(A) tail and its associated poly(A)-binding protein (Pab1p) are important regulators of gene expression. One role for this complex in the yeast Saccharomyces cerevisiae is in translation initiation through an interaction with a 115-amino-acid region of the translation initiation factor eIF4G. The eIF4G-interacting domain of Pab1p was mapped to its second RNA recognition motif (RRM2) in an in vitro binding assay. Moreover, RRM2 of Pab1p was required for poly(A) tail-dependent translation in yeast extracts. An analysis of a site-directed Pab1p mutation which bound to eIF4G but did not stimulate translation of uncapped, polyadenylated mRNA suggested additional Pab1p dependent events during translation initiation. These results support the model that the association of RRM2 of yeast Pab1p with eIF4G is a prerequisite for the poly(A) tail to stimulate the translation of mRNA in vitro. PMID- 9418853 TI - Potential Alu function: regulation of the activity of double-stranded RNA activated kinase PKR. AB - Cell stress, viral infection, and translational inhibition increase the abundance of human Alu RNA, suggesting that the level of these transcripts is sensitive to the translational state of the cell. To determine whether Alu RNA functions in translational homeostasis, we investigated its role in the regulation of double stranded RNA-activated kinase PKR. We found that overexpression of Alu RNA by cotransient transfection increased the expression of a reporter construct, which is consistent with an inhibitory effect on PKR. Alu RNA formed stable, discrete complexes with PKR in vitro, bound PKR in vivo, and antagonized PKR activation both in vitro and in vivo. Alu RNAs produced by either overexpression or exposure of cells to heat shock bound PKR, whereas transiently overexpressed Alu RNA antagonized virus-induced activation of PKR in vivo. Cycloheximide treatment of cells decreased PKR activity, coincident with an increase in Alu RNA. These observations suggest that the increased levels of Alu RNAs caused by cellular exposure to different stresses regulate protein synthesis by antagonizing PKR activation. This provides a functional role for mammalian short interspersed elements, prototypical junk DNA. PMID- 9418854 TI - Multiple roles for the MyoD basic region in transmission of transcriptional activation signals and interaction with MEF2. AB - Establishment of skeletal muscle lineages is controlled by the MyoD family of basic helix-loop-helix (bHLH) transcription factors. The ability of these factors to initiate myogenesis is dependent on two conserved amino acid residues, alanine and threonine, in the basic domains of these factors. It has been postulated that these two residues may be responsible for the initiation of myogenesis via interaction with an essential myogenic cofactor. The myogenic bHLH proteins cooperatively activate transcription and myogenesis through protein-protein interactions with members of the myocyte enhancer factor 2 (MEF2) family of MADS domain transcription factors. MyoD-E12 heterodimers interact with MEF2 proteins to synergistically activate myogenesis, while homodimers of E12, which lack the conserved alanine and threonine residues in the basic domain, do not interact with MEF2. We have examined whether the myogenic alanine and threonine in the MyoD basic region are required for interaction with MEF2. Here, we show that substitution of the MyoD basic domain with that of E12 does not prevent interaction with MEF2. Instead, the inability of alanine-threonine mutants of MyoD to initiate myogenesis is due to a failure to transmit transcriptional activation signals provided either from the MyoD or the MEF2 activation domain. This defect in transcriptional transmission can be overcome by substitution of the MyoD or the MEF2 activation domain with the VP16 activation domain. These results demonstrate that myogenic bHLH-MEF2 interaction can be uncoupled from transcriptional activation and support the idea that the myogenic residues in myogenic bHLH proteins are essential for transmission of a transcriptional activation signal. PMID- 9418855 TI - Activation of p38 mitogen-activated protein kinase by sodium salicylate leads to inhibition of tumor necrosis factor-induced IkappaB alpha phosphorylation and degradation. AB - Many actions of the proinflammatory cytokines tumor necrosis factor (TNF) and interleukin-1 (IL-1) on gene expression are mediated by the transcription factor NF-kappaB. Activation of NF-kappaB by TNF and IL-1 is initiated by the phosphorylation of the inhibitory subunit, IkappaB, which targets IkappaB for degradation and leads to the release of active NF-kappaB. The nonsteroidal anti inflammatory drug sodium salicylate (NaSal) interferes with TNF-induced NF-kappaB activation by inhibiting phosphorylation and subsequent degradation of the IkappaB alpha protein. Recent evidence indicated that NaSal activates the p38 mitogen-activated protein kinase (MAPK), raising the possibility that inhibition of NF-kappaB activation by NaSal is mediated by p38 MAPK. We now show that inhibition of TNF-induced IkappaB alpha phosphorylation and degradation by NaSal is prevented by treatment of cells with SB203580, a highly specific p38 MAPK inhibitor. Both p38 activation and inhibition of TNF-induced IkappaB alpha degradation were seen after only 30 s to 1 min of NaSal treatment. Induction of p38 MAPK activation and inhibition of TNF-induced IkappaB alpha degradation were demonstrated with pharmacologically achievable doses of NaSal. These findings provide evidence for a role of NaSal-induced p38 MAPK activation in the inhibition of TNF signaling and suggest a possible role for the p38 MAPK in the anti-inflammatory actions of salicylates. In addition, these results implicate the p38 MAPK as a possible negative regulator of TNF signaling that leads to NF kappaB activation. PMID- 9418857 TI - Gene conversion tracts from double-strand break repair in mammalian cells. AB - Mammalian cells are able to repair chromosomal double-strand breaks (DSBs) both by homologous recombination and by mechanisms that require little or no homology. Although spontaneous homologous recombination is rare, DSBs will stimulate recombination by 2 to 3 orders of magnitude when homology is provided either from exogenous DNA in gene-targeting experiments or from a repeated chromosomal sequence. Using a gene-targeting assay in mouse embryonic stem cells, we now investigate the effect of heterology on recombinational repair of DSBs. Cells were cotransfected with an endonuclease expression plasmid to induce chromosomal DSBs and with substrates containing up to 1.2% heterology from which to repair the DSBs. We find that heterology decreases the efficiency of recombinational repair, with 1.2% sequence divergence resulting in an approximately sixfold reduction in recombination. Gene conversion tract lengths were examined in 80 recombinants. Relatively short gene conversion tracts were observed, with 80% of the recombinants having tracts of 58 bp or less. These results suggest that chromosome ends in mammalian cells are generally protected from extensive degradation prior to recombination. Gene conversion tracts that were long (up to 511 bp) were continuous, i.e., they contained an uninterrupted incorporation of the silent mutations. This continuity suggests that these long tracts arose from extensive degradation of the ends or from formation of heteroduplex DNA which is corrected with a strong bias in the direction of the unbroken strand. PMID- 9418856 TI - A farnesyltransferase inhibitor induces tumor regression in transgenic mice harboring multiple oncogenic mutations by mediating alterations in both cell cycle control and apoptosis. AB - The farnesyltransferase inhibitor L-744,832 selectively blocks the transformed phenotype of cultured cells expressing a mutated H-ras gene and induces dramatic regression of mammary and salivary carcinomas in mouse mammary tumor virus (MMTV) v-Ha-ras transgenic mice. To better understand how the farnesyltransferase inhibitors might be used in the treatment of human tumors, we have further explored the mechanisms by which L-744,832 induces tumor regression in a variety of transgenic mouse tumor models. We assessed whether L-744,832 induces apoptosis or alterations in cell cycle distribution and found that the tumor regression in MMTV-v-Ha-ras mice could be attributed entirely to elevation of apoptosis levels. In contrast, treatment with doxorubicin, which induces apoptosis in many tumor types, had a minimal effect on apoptosis in these tumors and resulted in a less dramatic tumor response. To determine whether functional p53 is required for L 744,832-induced apoptosis and the resultant tumor regression, MMTV-v-Ha-ras mice were interbred with p53(-/-) mice. Tumors in ras/p53(-/-) mice treated with L 744,832 regressed as efficiently as MMTV-v-Ha-ras tumors, although this response was found to be mediated by both the induction of apoptosis and an increase in G1 with a corresponding decrease in the S-phase fraction. MMTV-v-Ha-ras mice were also interbred with MMTV-c-myc mice to determine whether ras/myc tumors, which possess high levels of spontaneous apoptosis, have the potential to regress through a further increase in apoptosis levels. The ras/myc tumors were found to respond nearly as efficiently to L-744,832 treatment as the MMTV-v-Ha-ras tumors, although no induction of apoptosis was observed. Rather, the tumor regression in the ras/myc mice was found to be mediated by a large reduction in the S-phase fraction. In contrast, treatment of transgenic mice harboring an activated MMTV-c neu gene did not result in tumor regression. These results demonstrate that a farnesyltransferase inhibitor can induce regression of v-Ha-ras-bearing tumors by multiple mechanisms, including the activation of a suppressed apoptotic pathway, which is largely p53 independent, or by cell cycle alterations, depending upon the presence of various other oncogenic genetic alterations. PMID- 9418858 TI - Activation of beta-globin promoter by erythroid Kruppel-like factor. AB - Erythroid Kruppel-like factor (EKLF), an erythroid tissue-specific Kruppel-type zinc finger protein, binds to the beta-globin gene CACCC box and is essential for beta-globin gene expression. EKLF does not activate the gamma gene, the CACCC sequence of which differs from that of the beta gene. To test whether the CACCC box sequence difference is the primary determinant of the selective activation of the beta gene by EKLF, the CACCC boxes of beta and gamma genes were swapped and the resulting promoter activities were assayed by transient transfections in CV-1 cells. EKLF activated the beta promoter carrying a gamma CACCC box at a level comparable to that at which it activated the wild-type beta promoter, whereas EKLF failed to activate a gamma promoter carrying the beta CACCC box, despite the presence of the optimal EKLF binding site. Similar results were obtained in K562 cells. The possibility that overexpressed EKLF superactivated the beta promoter carrying the gamma CACCC box, or that EKLF activated the mutated beta promoter through the intact distal CACCC box, was excluded. To test whether the position of the CACCC box in the beta or gamma promoter determined EKLF specificity, the proximal beta CACCC box sequence was created at the position of the beta promoter (-140) which corresponds to the position of the CACCC box on the gamma promoter. Similarly, the beta CACCC box was created in the position of the gamma promoter ( 90) corresponding to the position of the CACCC box in the beta promoter. EKLF retained weak activation potential on the beta(-140CAC) promoter, whereas EKLF failed to activate the gamma(-90betaCAC) promoter even though that promoter contained an optimal EKLF binding site at the optimal position. Taken together, our findings indicate that the specificity of the activation of the beta promoter by EKLF is determined by the overall structure of the beta promoter rather than solely by the sequence of the beta gene CACCC box. PMID- 9418859 TI - Transforming growth factor beta stimulates the human immunodeficiency virus 1 enhancer and requires NF-kappaB activity. AB - Transforming growth factor beta (TGF-beta) is the prototype of a large superfamily of signaling molecules involved in the regulation of cell growth and differentiation. In certain patients infected with human immunodeficiency virus type 1 (HIV-1), increased levels of TGF-beta promoted the production of virus and also impaired the host immune system. In an effort to understand the signaling events linking TGF-beta action and HIV production, we show here that TGF-beta can stimulate transcription from the HIV-1 long terminal repeat (LTR) promoter through NF-kappaB binding sites in both HaCaT and 300.19 pre-B cells. When introduced into a minimal promoter, NF-kappaB binding sites supported nearly 30 fold activation from the luciferase reporter upon TGF-beta treatment. Electrophoretic mobility shift assay indicated that a major factor binding to the NF-kappaB site is the p50-p65 heterodimeric NF-kappaB in HaCaT cells. Coexpression of Gal4-p65 chimeric proteins supported TGF-beta ligand-dependent gene expression from a luciferase reporter gene driven by Gal4 DNA binding sites. NF-kappaB activity present in HaCaT cells was not affected by TGF-beta treatment as judged by the unchanged DNA binding activity and concentrations of p50 and p65 proteins. Consistently, steady-state levels of IkappaB alpha and IkappaB beta proteins were not changed by TGF-beta treatment. Our results demonstrate that TGF beta is able to stimulate transcription from the HIV-1 LTR promoter by activating NF-kappaB through a mechanism distinct from the classic NF-kappaB activation mechanism involving the degradation of IkappaB proteins. PMID- 9418860 TI - The oncogenic capacity of HRX-ENL requires the transcriptional transactivation activity of ENL and the DNA binding motifs of HRX. AB - The HRX gene (also called MLL, ALL-1, and Htrx) at chromosome band 11q23 is associated with specific subsets of acute leukemias through translocations that result in its fusion with a variety of heterologous partners. Two of these partners, ENL and AF9, code for proteins that are highly similar to each other and as fusions with HRX induce myeloid leukemias in mice as demonstrated by retroviral gene transfer and knock-in experiments, respectively. In the present study, a structure-function analysis was performed to determine the molecular requirements for in vitro immortalization of murine myeloid cells by HRX-ENL. Deletions of either the AT hook motifs or the methyltransferase homology domain of HRX substantially impaired the transforming effects of HRX-ENL. The methyltransferase homology domain was shown to bind non-sequence specifically to DNA in vitro, providing evidence that the full transforming activity of HRX-ENL requires multiple DNA binding structures in HRX. The carboxy-terminal 84 amino acids of ENL, which encode two predicted helical structures highly conserved in AF9, were necessary and sufficient for transformation when they were fused to HRX. Similarly, mutations that deleted one or both of these conserved helices completely abrogated the transcriptional activation properties of ENL. This finding correlates, for the first time, a biological function of an HRX fusion partner with the transforming activity of the chimeric proteins. Our studies support a model in which HRX-ENL induces myeloid transformation by deregulating subordinate genes through a gain of function contributed by the transcriptional effector properties of ENL. PMID- 9418861 TI - Myotonic dystrophy kinase-related Cdc42-binding kinase acts as a Cdc42 effector in promoting cytoskeletal reorganization. AB - The Rho GTPases play distinctive roles in cytoskeletal reorganization associated with growth and differentiation. The Cdc42/Rac-binding p21-activated kinase (PAK) and Rho-binding kinase (ROK) act as morphological effectors for these GTPases. We have isolated two related novel brain kinases whose p21-binding domains resemble that of PAK whereas the kinase domains resemble that of myotonic dystrophy kinase related ROK. These approximately 190-kDa myotonic dystrophy kinase-related Cdc42 binding kinases (MRCKs) preferentially phosphorylate nonmuscle myosin light chain at serine 19, which is known to be crucial for activating actin-myosin contractility. The p21-binding domain binds GTP-Cdc42 but not GDP-Cdc42. The multidomain structure includes a cysteine-rich motif resembling those of protein kinase C and n-chimaerin and a putative pleckstrin homology domain. MRCK alpha and Cdc42V12 colocalize, particularly at the cell periphery in transfected HeLa cells. Microinjection of plasmid encoding MRCK alpha resulted in actin and myosin reorganization. Expression of kinase-dead MRCK alpha blocked Cdc42V12-dependent formation of focal complexes and peripheral microspikes. This was not due to possible sequestration of the p21, as a kinase-dead MRCK alpha mutant defective in Cdc42 binding was an equally effective blocker. Coinjection of MRCK alpha plasmid with Cdc42 plasmid, at concentrations where Cdc42 plasmid by itself elicited no effect, led to the formation of the peripheral structures associated with a Cdc42-induced morphological phenotype. These Cdc42-type effects were not promoted upon coinjection with plasmids of kinase-dead or Cdc42-binding-deficient MRCK alpha mutants. These results suggest that MRCK alpha may act as a downstream effector of Cdc42 in cytoskeletal reorganization. PMID- 9418862 TI - Mutations of the Drosophila dDP, dE2F, and cyclin E genes reveal distinct roles for the E2F-DP transcription factor and cyclin E during the G1-S transition. AB - Activation of heterodimeric E2F-DP transcription factors can drive the G1-S transition. Mutation of the Drosophila melanogaster dE2F gene eliminates transcriptional activation of several replication factors at the G1-S transition and compromises DNA replication. Here we describe a mutation in the Drosophila dDP gene. As expected for a defect in the dE2F partner, this mutation blocks G1-S transcription of DmRNR2 and cyclin E as previously described for mutations of dE2F. Mutation of dDP also causes an incomplete block of DNA replication. When S phase is compromised by reducing the activity of dE2F-dDP by either a dE2F or dDP mutation, the first phenotype detected is a reduction in the intensity of BrdU incorporation and a prolongation of the labeling. Notably, in many cells, there was no detected delay in entry into this compromised S phase. In contrast, when cyclin E function was reduced by a hypomorphic allele combination, BrdU incorporation was robust but the timing of S-phase entry was delayed. We suggest that dE2F-dDP contributes to the expression of two classes of gene products: replication factors, whose abundance has a graded effect on replication, and cyclin E, which triggers an all-or-nothing transition from G1 to S phase. PMID- 9418863 TI - Phosphorylation of Enabled by the Drosophila Abelson tyrosine kinase regulates the in vivo function and protein-protein interactions of Enabled. AB - Drosophila Enabled (Ena) is a member of a family of cytoskeleton-associated proteins including mammalian vasodilator-stimulated phosphoprotein and murine Enabled that regulate actin cytoskeleton assembly. Mutations in Drosophila ena were discovered as dominant genetic suppressors of mutations in the Abelson tyrosine kinase (Abl), suggesting that Ena and Abl function in the same pathway or process. We have identified six tyrosine residues on Ena that are phosphorylated by Abl in vitro and in vivo. Mutation of these phosphorylation sites to phenylalanine partially impaired the ability of Ena to restore viability to ena mutant animals, indicating that phosphorylation is required for optimal Ena function. Phosphorylation of Ena by Abl inhibited the binding of Ena to SH3 domains in vitro, suggesting that one effect of Ena phosphorylation may be to modulate its association with other proteins. PMID- 9418864 TI - Structural determinants of SHP-2 function and specificity in Xenopus mesoderm induction. AB - SHP-2 is a positive component of many receptor tyrosine kinase signaling pathways. The related protein-tyrosine phosphatase (PTP) SHP-1 usually acts as a negative regulator. The precise domains utilized by SHP-2 to transmit positive signals in vivo and the basis for specificity between SHP-1 and SHP-2 are not clear. In Xenopus, SHP-2 is required for mesoderm induction and completion of gastrulation. We investigated the effects of SHP-2 mutants and SHP-2/SHP-1 chimeras on basic fibroblast growth factor-induced mesoderm induction. Both SH2 domains and the PTP domain are required for normal SHP-2 function in this pathway. The N-terminal SH2 domain is absolutely required, whereas the C-terminal SH2 contributes to wild-type function. The C-terminal tyrosyl phosphorylation sites and proline-rich region are dispensable, arguing against adapter models of SHP-2 function. Although the SH2 domains contribute to SHP-2 specificity, studies of SHP chimeras reveal that substantial specificity resides in the PTP domain. Thus, PTP domains exhibit biologically relevant specificity in vivo, and noncatalytic and catalytic domains of PTPs contribute to specificity in a combinatorial fashion. PMID- 9418866 TI - Functional dissection of YA, an essential, developmentally regulated nuclear lamina protein in Drosophila melanogaster. AB - The Drosophila YA protein is a nuclear lamina component whose function is essential to initiate embryonic development. To identify regions of YA required for its action in its normal cellular context, we made targeted mutations in the YA protein and tested their consequences in flies and embryos in vivo. We found that critical amino acids are distributed along the length of the YA molecule, with functionally important regions including the N- and the C-terminal ends, the cysteine residues in YA's two potential zinc fingers, a serine/threonine-rich region, and a potential maturation-promoting factor or mitogen-activated protein kinase phosphorylation target site, ITPIR. In addition, several Ya mutations showed intragenic complementation, with N-terminal mutations complementing C terminal mutations, suggesting that YA proteins interact with one another. In support of this interaction, we demonstrated by immunoprecipitation that YA molecules are present in complexes with each other. Finally, we showed that the C terminal 179 amino acids of YA are necessary to target, or retain, YA in the nuclear envelope. PMID- 9418867 TI - Frequent loss of the active site during variant surface glycoprotein expression site switching in vitro in Trypanosoma brucei. AB - African trypanosomes undergo antigenic variation of their variant surface glycoprotein (VSG) coat to avoid being killed by their mammalian hosts. The active VSG gene is located in one of many telomeric expression sites. Replacement of the VSG gene in the active site or switching between expression sites can give rise to a new VSG coat. To study Trypanosoma brucei VSG expression site inactivation rather than VSG gene switching, it is useful to have an in vitro negative-selection system independent of the VSG. We have achieved this aim by using a viral thymidine kinase (TK) gene. Following integration of the TK gene downstream of the 221a VSG expression site promoter, transformant cell lines became sensitive to the nucleoside analog 1-(2-deoxy-2-fluoro-8-D arabinofuranosyl)-5-iodouracil. These TK trypanosomes were able to revert to resistance at a rate approaching 10(-5) per cell per generation. The majority of revertants expressed a new VSG gene even though there had been no selection against the VSG itself. Analysis of these switched variants showed that some had shut down TK expression via an in situ expression site switch. However, most variants had the complete 221 expression site deleted and another VSG expression site activated. We speculate that a new VSG expression site cannot switch on without inactivation of the old site. PMID- 9418865 TI - Characterization of the mitochondrial inner membrane translocase complex: the Tim23p hydrophobic domain interacts with Tim17p but not with other Tim23p molecules. AB - Tim23p is a mitochondrial inner membrane protein essential for the import of proteins from the cytosol. Tim23p contains an amino-terminal hydrophilic segment and a carboxyl-terminal hydrophobic domain (Tim23Cp). To study the functions and interactions of the two parts of Tim23p separately, we constructed tim23N, encoding only the hydrophilic region of Tim23p, and tim23C, encoding only the hydrophobic domain of Tim23p. Only the Tim23C protein is imported into mitochondria, indicating that the mitochondrial targeting information in Tim23p resides in its membrane spans or intervening loops. Tim23Cp, however, cannot substitute for full-length Tim23p, suggesting that the hydrophilic portion of Tim23p also performs an essential function in mitochondrial protein import. We found that overexpression of Tim23Cp is toxic to yeast cells that carry the tim23 1 mutation. Excess Tim23Cp causes Tim23-1p to disappear, leaving tim23-1 cells without a full-length version of the Tim23 protein. If Tim17p, another inner membrane import component, is overexpressed along with Tim23Cp, the toxicity of Tim23Cp is largely reversed and the Tim23-1 protein no longer disappears. In coimmunoprecipitations from solubilized mitochondria, Tim17p associates with the Tim23C protein. In addition, we show that Tim23p and Tim17p can be chemically cross-linked to each other in intact mitochondria. We conclude that the hydrophobic domain encoded by tim23C targets Tim23p to the mitochondria and mediates the direct interaction between Tim23p and Tim17p. In contrast, Tim23Cp cannot be coimmunoprecipitated with Tim23p, raising the possibility that the hydrophobic domain of Tim23p does not interact with other Tim23 molecules. PMID- 9418868 TI - p130 is dispensable in peripheral T lymphocytes: evidence for functional compensation by p107 and pRB. AB - The proteins encoded by the retinoblastoma gene family, pRB, p107, and p130, have been implicated in the regulation of cellular proliferation, differentiation, and transformation. Because interactions between p130 and E2F transcription factors have been proposed to play a role in the establishment and/or maintenance of quiescence in human peripheral T lymphocytes, we examined lymphoid differentiation and proliferation in p130-deficient mice. We show that p130-/- T cells proliferate normally in culture and exhibit normal cell-mediated immune function in vivo. However, p130-/- T lymphocytes expressed elevated levels of p107, and the characteristic p130-E2F DNA binding complex was replaced by a p107 E2F complex. Adoptive transfer of fetal liver lymphoid progenitors allowed us to circumvent the neonatal lethality associated with loss of p130 and p107 and to analyze the phenotype of p130-/-;p107-/- peripheral T lymphocytes. These cells achieved a quiescent state, exhibited derepression of a subset of E2F target genes, and were hypersensitive to concanavalin A stimulation. Interestingly, a significant portion of the E2F-4 in p130-/-;p107-/- T cells was detected in a complex with pRB and an as-yet-unidentified protein. These findings provide a biochemical basis for functional compensation between pRB family proteins. PMID- 9418869 TI - Dual sets of chimeric alleles identify specificity sequences for the bE and bW mating and pathogenicity genes of Ustilago maydis. AB - The b mating-type locus of the fungal plant pathogen Ustilago maydis encodes two multiallelic gene products, bE and bW, that control the formation and maintenance of the infectious cell type. Dimerization via the N-terminal regions of bE and bW proteins encoded by alleles of different specificities establishes a homeodomain containing transcription factor. The bE and bW products encoded by alleles of like specificities fail to dimerize. We constructed sets of chimeric alleles for the bE1 and bE2 genes and for the bW1 and bW2 genes to identify sequences that control specificity. The mating behavior of strains carrying chimeric alleles identified three classes of specificity: b2 (class I), specificity different from either parental type (class II), and b1 (class III). Crosses between strains carrying bE and bW chimeric alleles identified two short blocks of amino acids that influence specificity and that are located in the N-terminal variable regions of the b proteins. Comparisons of pairs of chimeric alleles encoding polypeptides differing in specificity and differing at single amino acid positions identified 16 codon positions that influence the interaction between bE and bW. Fifteen of these positions lie within the blocks of amino acids identified by crosses between the strains carrying chimeric alleles. Overall, this work provides insight into the organization of the regions that control recognition. PMID- 9418870 TI - CIF150, a human cofactor for transcription factor IID-dependent initiator function. AB - The transcription factor IID (TFIID) complex is highly conserved between the Drosophila and mammalian systems. A mammalian homolog has been described for all the Drosophila TATA box-binding protein-associated factors (TAFs), with the exception of dTAF(II)150. We previously reported the identification of CIF, an essential cofactor for TFIID-dependent transcription from promoters containing initiator (Inr) elements. Here we describe the molecular cloning of CIF150, the human homolog of dTAF(II)150, and present biochemical evidence that this factor is involved in Inr activity. CIF150 is capable of mediating TFIID-dependent Inr activity in a complementation assay, and a protein fraction lacking Inr activity lacks detectable amounts of CIF150. Despite the striking similarity to dTAF(II)150, CIF150 does not appear to be associated with human TFIID. However, in vitro binding assays revealed a specific and direct interaction between CIF150 and hTAF(II)135. This interaction might be structurally important for the functional interaction between CIF150 and human TFIID, since CIF150 stabilizes TFIID binding to a core promoter. PMID- 9418871 TI - DDB, a putative DNA repair protein, can function as a transcriptional partner of E2F1. AB - The transcription factor E2F1 is believed to be involved in the regulated expression of the DNA replication genes. To gain insights into the transcriptional activation function of E2F1, we looked for proteins in HeLa nuclear extracts that bind to the activation domain of E2F1. Here we show that DDB, a putative DNA repair protein, associates with the activation domain of E2F1. DDB was identified as a heterodimeric protein (48 and 127 kDa) that binds to UV-damaged DNA. We show that the UV-damaged-DNA binding activity from HeLa nuclear extracts can associate with the activation domain of E2F1. Moreover, the 48-kDa subunit of DDB, synthesized in vitro, binds to a fusion protein of E2F1 depending on the C-terminal activation domain. The interaction between DDB and E2F1 can also be detected by coimmunoprecipitation experiments. Immunoprecipitation of an epitope-tagged DDB from cell extracts resulted in the coprecipitation of E2F1. In a reciprocal experiment, immunoprecipitates of E2F1 were found to contain DDB. Fractionation of HeLa nuclear extracts also revealed a significant overlap in the elution profiles of E2F1 and DDB. For instance, DDB, which does not bind to the E2F sites, was enriched in the high-salt fractions containing E2F1 during chromatography through an E2F-specific DNA affinity column. We also observed evidence for a functional interaction between DDB and E2F1 in living cells. For instance, expression of DDB specifically stimulated E2F1-activated transcription. In addition, the transcriptional activation function of a heterologous transcription factor containing the activation domain of E2F1 was stimulated by coexpression of DDB. Moreover, DDB expression could overcome the retinoblastoma protein (Rb)-mediated inhibition of E2F1-activated transcription. The results suggest that this damaged-DNA binding protein can function as a transcriptional partner of E2F1. We speculate that the damaged-DNA binding function of DDB, besides repair, might serve as a negative regulator of E2F1-activated transcription, as damaged DNA will sequester DDB and make it unavailable for E2F1. Furthermore, the binding of DDB to damaged DNA might be involved in downregulating the replication genes during growth arrest induced by damaged DNA. PMID- 9418872 TI - Transformation suppression by protein tyrosine phosphatase 1B requires a functional SH3 ligand. AB - We have recently shown that protein tyrosine phosphatase 1B (PTP1B) associates with the docking protein p130Cas in 3Y1 rat fibroblasts. This interaction is mediated by a proline-rich sequence on PTP1B and the SH3 domain on p130Cas. Expression of wild-type PTP1B (WT-PTP1B), but not a catalytically competent, proline-to-alanine point mutant that cannot bind p130Cas (PA-PTP1B), causes substantial tyrosine dephosphorylation of p130Cas (F. Liu, D. E. Hill, and J. Chernoff, J. Biol. Chem. 271:31290-31295, 1996). Here we demonstrate that WT-, but not PA-PTP1B, inhibits transformation of rat 3Y1 fibroblasts by v-crk, -src, and -ras, but not by v-raf. These effects on transformation correlate with the phosphorylation status of p130Cas and two proteins that are associated with p130Cas, Paxillin and Fak. Expression of WT-PTP1B reduces formation of p130Cas Crk complexes and inhibits mitogen-activated protein kinase activation by Src and Crk. These data show that transformation suppression by PTP1B requires a functional SH3 ligand and suggest that p130Cas may represent an important physiological target of PTP1B in cells. PMID- 9418873 TI - A novel mre11 mutation impairs processing of double-strand breaks of DNA during both mitosis and meiosis. AB - Using complementation tests and nucleotide sequencing, we showed that the rad58-4 mutation was an allele of the MRE11 gene and have renamed the mutation mre11-58. Two amino acid changes from the wild-type sequence were identified; one is located at a conserved site of a phosphodiesterase motif, and the other is a homologous amino acid change at a nonconserved site. Unlike mre11 null mutations, the mre11-58 mutation allowed meiosis-specific double-strand DNA breaks (DSBs) to form at recombination hot spots but failed to process those breaks. DSB ends of this mutant were resistant to lambda exonuclease treatment. These phenotypes are similar to those of rad50S mutants. In contrast to rad50S, however, mre11-58 was highly sensitive to methyl methanesulfonate treatment. DSB end processing induced by HO endonuclease was suppressed in both mre11-58 and the mre11 disruption mutant. We constructed a new mre11 mutant that contains only the phosphodiesterase motif mutation of the Mre11-58 protein and named it mre11-58S. This mutant showed the same phenotypes observed in mre11-58, suggesting that the phosphodiesterase consensus sequence is important for nucleolytic processing of DSB ends during both mitosis and meiosis. PMID- 9418874 TI - Telomere variation in Xenopus laevis. AB - Eukaryotic telomeres are variable at several levels, from the length of the simple sequence telomeric repeat tract in different cell types to the presence or number of telomere-adjacent DNA sequence elements in different strains or individuals. We have investigated the sequence organization of Xenopus laevis telomeres by use of the vertebrate telomeric repeat (TTAGGG)n and blot hybridization analysis. The (TTAGGG)n-hybridizing fragments, which ranged from less than 10 to over 50 kb with frequently cutting enzymes, defined a pattern that was polymorphic between individuals. BAL 31 exonuclease treatment confirmed that these fragments were telomeric. The polymorphic fragments analyzed did not hybridize to 5S RNA sequences, which are telomeric according to in situ hybridization. When telomeric fragments from offspring (whole embryos) were compared to those from the spleens of the parents, the inheritance pattern of some bands was found to be unusual. Furthermore, in one cross, the telomeres of the embryo were shorter than the telomeres of the parents' spleen, and in another, the male's testis telomeres were shorter than those of the male's spleen. Our data are consistent with a model for chromosome behavior that involves a significant amount of DNA rearrangement at telomeres and suggest that length regulation of Xenopus telomeres is different from that observed for Mus spretus and human telomeres. PMID- 9418875 TI - Poly(A)-driven and poly(A)-assisted termination: two different modes of poly(A) dependent transcription termination. AB - We mapped the elements that mediate termination of transcription downstream of the chicken betaH- and betaA-globin gene poly(A) sites. We found no unique element and no segment of 3'-flanking DNA to be significantly more effective than any other. When we replaced the native 3'-flanking DNA with bacterial DNA, it too supported transcription termination. Termination in the bacterial DNA depended on a functional poly(A) signal, which apparently compelled termination to occur in the downstream DNA with little regard for its sequence. We also studied premature termination by poorly processive polymerases close to the promoter. The rate of premature termination varied for different DNA sequences. However, the efficiencies of poly(A)-driven termination and promoter-proximal premature termination varied similarly on different DNAs, suggesting that poly(A)-driven termination functions by returning the transcription complex to a form which resembles a prior state of low processivity. The poly(A)-driven termination described here differs dramatically from the poly(A)-assisted termination previously described for the simian virus 40 (SV40) early transcription unit. In the SV40 early transcription unit, essentially no termination occurs downstream of the poly(A) site unless a special termination element is present. The difference between the betaH-globin and SV40 modes of termination is governed by sequences in the upstream DNA. For maximum efficiency, the betaH-globin poly(A) signal required the assistance of upstream enhancing sequences. Moreover, the SV40 early poly(A) signal also drove termination in betaH-globin style when it was placed in a betaH-globin sequence context. These studies were facilitated by a rapid, improved method of run-on transcription analysis, based on the use of a vector containing two G-free cassettes. PMID- 9418876 TI - Isolation and characterization of new alleles of the cyclin-dependent kinase gene CDC28 with cyclin-specific functional and biochemical defects. AB - The G1 cyclin Cln2 negatively regulates the mating-factor pathway. In a genetic screen to identify factors required for this regulation, we identified an allele of CDC28 (cdc28-csr1) that blocked this function of Cln2. Cln2 immunoprecipitated from cdc28-csr1 cells was completely defective in histone H1 kinase activity, due to defects in Cdc28 binding and activation by Cln2. In contrast, Clb2-associated H1 kinase and Cdc28 binding was normal in immunoprecipitates from these cells. cdc28-csr1 was significantly deficient in other aspects of genetic interaction with Cln2. The cdc28-csr1 mutation was determined to be Q188P, in the T loop distal to most of the probable Cdk-cyclin interaction regions. We performed random mutagenesis of CDC28 to identify additional alleles incapable of causing CLN2-dependent mating-factor resistance but capable of complementing cdc28 temperature-sensitive and null alleles. Two such mutants had highly defective Cln2-associated kinase, but, surprisingly, two other mutants had levels of Cln2 associated kinase near to wild-type levels. We performed a complementary screen for CDC28 mutants that could cause efficient Cln2-dependent mating-factor resistance but not complement a cdc28 null allele. Most such mutants were found to alter residues essential for kinase activity; the proteins had little or no associated kinase activity in bulk or in association with Cln2. Several of these mutants also functioned in another assay for CLN2-dependent function not involving the mating-factor pathway, complementing the temperature sensitivity of a cln1 cln3 cdc28-csr1 strain. These results could indicate that Cln2-Cdc28 kinase activity is not directly relevant to some CLN2-mediated functions. Mutants of this sort should be useful in differentiating the function of Cdc28 complexed with different cyclin regulatory subunits. PMID- 9418877 TI - Regulation of transforming growth factor alpha expression in a growth factor independent cell line. AB - Aberrant transcriptional regulation of transforming growth factor alpha (TGF alpha) appears to be an important contributor to the malignant phenotype and the growth factor independence with which malignancy is frequently associated. However, little is known about the molecular mechanisms responsible for dysregulation of TGF alpha expression in the malignant phenotype. In this paper, we report on TGF alpha promoter regulation in the highly malignant growth factor independent cell line HCT116. The HCT116 cell line expresses TGF alpha and the epidermal growth factor receptor (EGFR) but is not growth inhibited by antibodies to EGFR or TGF alpha. However, constitutive expression of TGF alpha antisense RNA in the HCT116 cell line resulted in the isolation of clones with markedly reduced TGF alpha mRNA and which were dependent on exogenous growth factors for proliferation. We hypothesized that if TGF alpha autocrine activation is the major stimulator of TGF alpha expression in this cell line, TGF alpha promoter activity should be reduced in the antisense TGF alpha clones in the absence of exogenous growth factor. This was the case. Moreover, transcriptional activation of the TGF alpha promoter was restored in an antisense-TGF alpha-mRNA-expressing clone which had reverted to a growth factor-independent phenotype. Using this model system, we were able to identify a 25-bp element within the TGF alpha promoter which conferred TGF alpha autoregulation to the TGF alpha promoter in the HCT116 cell line. In the TGF alpha-antisense-RNA-expressing clones, this element was activated by exogenous EGF. This 25-bp sequence contained no consensus sequences of known transcription factors so that the TGF alpha or EGF regulatory element within this 25-bp sequence represents a unique element. Further characterization of this 25-bp DNA sequence by deletion analysis revealed that regulation of TGF alpha promoter activity by this sequence is complex, as both repressors and activators bind in this region, but the overall expression of the activators is pivotal in determining the level of response to EGF or TGF alpha stimulation. The specific nuclear proteins binding to this region are also regulated in an autocrine-TGF alpha-dependent fashion and by exogenous EGF in EGF deprived TGF alpha antisense clone 33. This regulation is identical to that seen in the growth factor-dependent cell line FET, which requires exogenous EGF for optimal growth. Moreover, the time response of the stimulation of trans-acting factor binding by EGF suggests that the effect is directly due to growth factor and not mediated by changes in growth state. We conclude that this element appears to represent the major positive regulator of TGF alpha expression in the growth factor-independent HCT116 cell line and may represent the major site of transcriptional dysregulation of TGF alpha promoter activity in the growth factor independent phenotype. PMID- 9418878 TI - A PEST-like sequence mediates phosphorylation and efficient ubiquitination of yeast uracil permease. AB - Uptake of uracil by the yeast Saccharomyces cerevisiae is mediated by a specific permease encoded by the FUR4 gene. Uracil permease located at the cell surface is subject to two covalent modifications: phosphorylation and ubiquitination. The ubiquitination step is necessary prior to permease endocytosis and subsequent vacuolar degradation. Here, we demonstrate that a PEST-like sequence located within the cytoplasmic N terminus of the protein is essential for uracil permease turnover. Internalization of the transporter was reduced when some of the serines within the region were converted to alanines and severely impaired when all five serines within the region were mutated or when this region was absent. The phosphorylation and degree of ubiquitination of variant permeases were inversely correlated with the number of serines replaced by alanines. A serine-free version of this sequence was very poorly phosphorylated, and elimination of this sequence prevented ubiquitination. Thus, it appears that the serine residues in the PEST like sequence are required for phosphorylation and ubiquitination of uracil permease. A PEST-like sequence in which the serines were replaced by glutamic acids allowed efficient permease turnover, suggesting that the PEST serines are phosphoacceptors. PMID- 9418880 TI - A novel functional human eukaryotic translation initiation factor 4G. AB - Mammalian eukaryotic translation initiation factor 4F (eIF4F) is a cap-binding protein complex consisting of three subunits: eIF4E, eIF4A, and eIF4G. In yeast and plants, two related eIF4G species are encoded by two different genes. To date, however, only one functional eIF4G polypeptide, referred to here as eIF4GI, has been identified in mammals. Here we describe the discovery and functional characterization of a closely related homolog, referred to as eIF4GII. eIF4GI and eIF4GII share 46% identity at the amino acid level and possess an overall similarity of 56%. The homology is particularly high in certain regions of the central and carboxy portions, while the amino-terminal regions are more divergent. Far-Western analysis and coimmunoprecipitation experiments were used to demonstrate that eIF4GII directly interacts with eIF4E, eIF4A, and eIF3. eIF4GII, like eIF4GI, is also cleaved upon picornavirus infection. eIF4GII restores cap-dependent translation in a reticulocyte lysate which had been pretreated with rhinovirus 2A to cleave endogenous eIF4G. Finally, eIF4GII exists as a complex with eIF4E in HeLa cells, because eIF4GII and eIF4E can be purified together by cap affinity chromatography. Taken together, our findings indicate that eIF4GII is a functional homolog of eIF4GI. These results may have important implications for the understanding of the mechanism of shutoff of host protein synthesis following picornavirus infection. PMID- 9418881 TI - A short sequence within two purine-rich enhancers determines 5' splice site specificity. AB - Purine-rich enhancers are exon sequences that promote inclusion of alternative exons, usually via activation of weak upstream 3' splice sites. A recently described purine-rich enhancer from the caldesmon gene has an additional activity by which it directs selection of competing 5' splice sites within an alternative exon. In this study, we have compared the caldesmon enhancer with another purine rich enhancer from the chicken cardiac troponin T (cTNT) gene for the ability to regulate flanking splice sites. Although similar in sequence and length, the two enhancers demonstrated strikingly different specificities towards 5' splice site choice when placed between competing 5' splice sites in an internal exon. The 32 nucleotide caldesmon enhancer caused effective usage of the exon-internal 5' splice site, whereas the 30-nucleotide cTNT enhancer caused effective usage of the exon-terminal 5' splice site. Both enhancer-mediated splicing pathways represented modulation of the default pathway in which both 5' splice sites were utilized. Each enhancer is multipartite, consisting of two purine-rich sequences of a simple (GAR)n repeat interdigitated with two enhancer-specific sequences. The entire enhancer was necessary for maximal splice site selectivity; however, a 5- to 7-nucleotide region from the 3' end of each enhancer dictated splice site selectivity. Mutations that interchanged this short region of the two enhancers switched specificity. The portion of the cTNT enhancer determinative for 5' splice site selectivity was different than that shown to be maximally important for activation of a 3' splice site, suggesting that enhancer environment can have a major impact on activity. These results are the first indication that individual purine-rich enhancers can differentiate between flanking splice sites. Furthermore, localization of the specificity of splice site choice to a short region within both enhancers indicates that subtle differences in enhancer sequence can have profound effects on the splicing pathway. PMID- 9418879 TI - The t(8;21) fusion product, AML-1-ETO, associates with C/EBP-alpha, inhibits C/EBP-alpha-dependent transcription, and blocks granulocytic differentiation. AB - AML-1B is a hematopoietic transcription factor that is functionally inactivated by multiple chromosomal translocations in human acute myeloblastic and B-cell lymphocytic leukemias. The t(8;21)(q22;q22) translocation replaces the C terminus, including the transactivation domain of AML-1B, with ETO, a nuclear protein of unknown function. We previously showed that AML-1-ETO is a dominant inhibitor of AML-1B-dependent transcriptional activation. Here we demonstrate that AML-1-ETO also inhibits C/EBP-alpha-dependent activation of the myeloid cell specific, rat defensin NP-3 promoter. AML-1B bound the core enhancer motifs present in the NP-3 promoter and activated transcription approximately sixfold. Similarly, C/EBP-alpha bound NP-3 promoter sequences and activated transcription approximately sixfold. Coexpression of C/EBP-alpha with AML-1B or its family members, AML-2 and murine AML-3, synergistically activated the NP-3 promoter up to 60-fold. The t(8;21) product, AML-1-ETO, repressed AML-1B-dependent activation of NP-3 and completely inhibited C/EBP-alpha-dependent activity as well as the synergistic activation. In contrast, the inv(16) product, which indirectly targets AML family members by fusing their heterodimeric DNA binding partner, CBF beta, to the myosin heavy chain, inhibited AML-1B but not C/EBP-alpha activation or the synergistic activation. AML-1-ETO and C/EBP-alpha were coimmunoprecipitated and thus physically interact in vivo. Deletion mutants demonstrated that the C terminus of ETO was required for AML-1-ETO-mediated repression of the synergistic activation but not for association with C/EBP alpha. Finally, overexpression of AML-1-ETO in myeloid progenitor cells prevented granulocyte colony-stimulating factor-induced differentiation. Thus, AML-1-ETO may contribute to leukemogenesis by specifically inhibiting C/EBP-alpha- and AML 1B-dependent activation of myeloid promoters and blocking differentiation. PMID- 9418882 TI - Yeast pre-mRNA splicing requires a pair of U1 snRNP-associated tetratricopeptide repeat proteins. AB - The U1 snRNP functions to nucleate spliceosome assembly on newly transcribed pre mRNA. Saccharomyces cerevisiae is unusual among eukaryotes in the greatly extended length of its U1 snRNA and the apparent increased polypeptide complexity of the corresponding U1 snRNP. In this paper, we report the identification of a novel U1 snRNP protein, Prp42p, with unexpected properties. Prp42p was identified by its surprising structural similarity to the essential U1 snRNP protein, Prp39p. Both Prp39p and Prp42p possess multiple copies of a variant tetratricopeptide repeat, an element implicated in a wide range of protein assembly events. Yeast strains depleted of Prp42p by transcriptional repression of a GAL1::PRP42 fusion gene arrest for splicing prior to pre-mRNA 5' splice site cleavage. Prp42p was not observed in a recent biochemical analysis of purified U1 snRNPs from S. cerevisiae (28). Nevertheless, antibodies directed against an epitope-tagged version of Prp42p specifically precipitate U1 snRNA from yeast extracts. Furthermore, Prp42p is required for U1 snRNP biogenesis, because yeast strains depleted of Prp42p formed incomplete U1 snRNPs that failed to produce stable complexes with pre-mRNA in vitro. The evidence shows that Prp39p and Prp42p are both required to configure the atypical yeast U1 snRNP into a structure compatible with its evolutionarily conserved role in pre-mRNA splicing. PMID- 9418883 TI - Heat shock factor increases the reinitiation rate from potentiated chromatin templates. AB - Transcription by RNA polymerase II is highly regulated at the level of initiation and elongation. Well-documented transcription activation mechanisms, such as the recruitment of TFIID and TFIIB, control the early phases of preinitiation complex formation. The heat shock genes provide an example for transcriptional regulation at a later step: in nuclei TFIID can be detected at the TATA box prior to heat induction. Using cell-free systems for chromatin reconstitution and transcription, we have analyzed the mechanisms by which heat shock factor (HSF) increases transcription of heat shock genes in chromatin. HSF affected transcription of naked DNA templates in multiple ways: (i) by speeding up the rate of preinitiation complex formation, (ii) by increasing the number of productive templates, and (iii) by increasing the reinitiation rate. Under the more physiological conditions of potentiated chromatin templates, HSF affected only the reinitiation rate. Activator-dependent reinitiation of transcription, obviating the slow assembly of the TFIID-TFIIA complex on a promoter, may be especially crucial for genes requiring a fast response to inducers. PMID- 9418884 TI - The large subunit of basal transcription factor SNAPc is a Myb domain protein that interacts with Oct-1. AB - The human RNA polymerase II and III snRNA promoters have similar enhancers, the distal sequence elements (DSEs), and similar basal promoter elements, the proximal sequence elements (PSEs). The DSE, which contains an octamer motif, binds broadly expressed activator Oct-1. The PSE binds a multiprotein complex referred to as SNAPc or PTF. On DNAs containing both an octamer site and a PSE, Oct-1 and SNAPc bind cooperatively. SNAPc consists of at least four stably associated subunits, SNAP43, SNAP45, SNAP50, and SNAP190. None of the three small subunits, which have all been cloned, can bind to the PSE on their own. Here we report the isolation of cDNAs corresponding to the largest subunit of SNAPc, SNAP190. SNAP190 contains an unusual Myb DNA binding domain consisting of four complete repeats (Ra to Rd) and a half repeat (Rh). A truncated protein consisting of the last two SNAP190 Myb repeats, Rc and Rd, can bind to the PSE, suggesting that the SNAP190 Myb domain contributes to recognition of the PSE by the SNAP complex. SNAP190 is required for snRNA gene transcription by both RNA polymerases II and III and interacts with SNAP45. In addition, SNAP190 interacts with Oct-1. Together, these results suggest that the largest subunit of the SNAP complex is involved in direct recognition of the PSE and is a target for the Oct 1 activator. They also provide an example of a basal transcription factor containing a Myb DNA binding domain. PMID- 9418885 TI - p16INK4A participates in a G1 arrest checkpoint in response to DNA damage. AB - Members of the INK4 protein family specifically inhibit cyclin-dependent kinase 4 (cdk4) and cdk6-mediated phosphorylation of the retinoblastoma susceptibility gene product (Rb). p16INK4A, a prototypic INK4 protein, has been identified as a tumor suppressor in many human cancers. Inactivation of p16INK4A in tumors expressing wild-type Rb is thought to be required in order for many malignant cell types to enter S phase efficiently or to escape senescence. Here, we demonstrate another mechanism of tumor suppression by implicating p16INK4A in a G1 arrest checkpoint in response to DNA damage. Calu-1 non-small cell lung cancer cells, which retain Rb and lack p53, do not arrest in G1 following DNA damage. However, engineered expression of p16INK4A at levels compatible with cell proliferation restores a G1 arrest checkpoint in response to treatment with gamma irradiation, topoisomerase I and II inhibitors, and cisplatin. A similar checkpoint can be demonstrated in p53-/- fibroblasts that express p16INK4A. DNA damage-induced G1 arrest, which requires the expression of pocket proteins such as Rb, can be abrogated by overexpression of cdk4, kinase-inactive cdk4 variants capable of sequestering p16INK4A, or a cdk4 variant incapable of binding p16INK4A. After exposure to DNA-damaging agents, there was no change either in overall levels of p16INK4A or in amounts of p16INK4A found in complex with cdks 4 and 6. Nonetheless, p16INK4A expression is required for the reduction in cdk4- and cdk6-mediated Rb kinase activity observed in response to DNA damage. During tumor progression, loss of p16INK4A expression may be necessary for cells with wild-type Rb to bypass this G1 arrest checkpoint and attain a fully transformed phenotype. PMID- 9418886 TI - Peroxisome targeting signal type 1 (PTS1) receptor is involved in import of both PTS1 and PTS2: studies with PEX5-defective CHO cell mutants. AB - To investigate the mechanisms of peroxisome assembly and the molecular basis of peroxisome assembly disorders, we isolated and characterized a peroxisome deficient CHO cell mutant, ZP139, which was found to belong to human complementation group II, the same group as that of our earlier mutant, ZP105. These mutants had a phenotypic deficiency in the import of peroxisomal targeting signal type 1 (PTS1) proteins. Amino-terminal extension signal (PTS2)-mediated transport, including that of 3-ketoacyl coenzyme A thiolase, was also defective in ZP105 but not in ZP139. PEX5 cDNA, encoding the PTS1 receptor (PTS1R), was isolated from wild-type CHO-K1 cells. PTS1R's deduced primary sequence comprised 595 amino acids, 7 amino acids less than the human homolog, and contained seven tetratricopeptide repeat (TPR) motifs at the C-terminal region. Chinese hamster PTS1R showed 94, 28, and 24% amino acid identity with PTS1Rs from humans, Pichia pastoris, and Saccharomyces cerevisiae, respectively. A PTS1R isoform (PTS1RL) with 632 amino acid residues was identified in CHO cells; for PTS1R, 37 amino acids were inserted between residues at positions 215 and 216 of a shorter isoform (PTS1RS). Southern blot analysis of CHO cell genomic DNA suggested that these two isoforms are derived from a single gene. Both types of PEX5 complemented impaired import of PTS1 in mutants ZP105 and ZP139. PTS2 import in ZP105 was rescued only by PTS1RL. This finding strongly suggests that PTS1RL is also involved in the transport of PTS2. Mutations in PEX5 were determined by reverse transcription-PCR: a G-to-A transition resulted in one amino acid substitution: Gly298Glu of PTS1RS (G335E of PTS1RL) in ZP105 and Gly485Glu of PTS1RS (G522E of PTS1RL) in ZP139. Both mutations were in the TPR domains (TPR1 and TPR6), suggesting the functional consequence of these domains in protein translocation. The implications of these mutations are discussed. PMID- 9418887 TI - Activation of the kexin from Schizosaccharomyces pombe requires internal cleavage of its initially cleaved prosequence. AB - Members of the kexin family of processing enzymes are responsible for the cleavage of many proproteins during their transport through the secretory pathway. The enzymes themselves are made as inactive precursors, and we investigated the activation process by studying the maturation of Krp1, a kexin from the fission yeast Schizosaccharomyces pombe. Using a cell-free translation translocation system prepared from Xenopus eggs, we found that Krp1 is made as a preproprotein that loses the presequence during translocation into the endoplasmic reticulum. The prosequence is also rapidly cleaved in a reaction that is autocatalytic and probably intramolecular and is inhibited by disruption of the P domain. Prosequence cleavage normally occurs at Arg-Tyr-Lys-Arg102/ (primary cleavage site) but can occur at Lys-Arg82 (internal cleavage site) and/or Trp-Arg99 when the basic residues are removed from the primary site. Cleavage of the prosequence is necessary but not sufficient for activation, and Krp1 is initially unable to process substrates presented in trans. Full activation is achieved after further incubation in the extract and is coincident with the addition of O-linked sugars. O glycosylation is not, however, essential for activity, and the crucial event appears to be cleavage of the initially cleaved prosequence at the internal site. Our results are consistent with a model in which the cleaved prosequence remains noncovalently associated with the catalytic domain and acts as an autoinhibitor of the enzyme. Inhibition is then relieved by a second (internal) cleavage of the inhibitory prosequence. Further support for this model is provided by our finding that overexpression of a Krp1 prosequence lacking a cleavable internal site dramatically reduced the growth rate of otherwise wild-type S. pombe cells, an effect that was not seen after overexpression of the normal, internally cleavable, prosequence or prosequences that lack the Lys-Arg102 residues. PMID- 9418888 TI - Opposite transcriptional effects of cyclic AMP-responsive elements in confluent or p27KIP-overexpressing cells versus serum-starved or growing cells. AB - The minute virus of mice, an autonomous parvovirus, requires entry of host cells into the S phase of the cell cycle for its DNA to be amplified and its genes expressed. This work focuses on the P4 promoter of this parvovirus, which directs expression of the transcription unit encoding the parvoviral nonstructural polypeptides. These notably include protein NS1, necessary for the S-phase dependent burst of parvoviral DNA amplification and gene expression. The activity of the P4 promoter is shown to be regulated in a cell cycle-dependent manner. At the G1/S-phase transition, the promoter is activated via a cis-acting DNA element which interacts with phase-specific complexes containing the cellular transcription factor E2F. It is inhibited, on the other hand, in cells arrested in G1 due to contact inhibition. This inhibitory effect is not observed in serum starved cells. It is mediated in cis by cyclic AMP response elements (CREs). Unlike serum-starved cells, confluent cells accumulate the cyclin-dependent kinase inhibitor p27, suggesting that the switch from CRE-mediated activation to CRE-mediated repression involves the p27 protein. Accordingly, plasmid-driven overexpression of p27 causes down-modulation of promoter P4 in growing cells, depending on the presence of at least two functional CREs. No such effect is observed with two other cyclin-dependent kinase inhibitors, p16 and p21. Given the importance of P4-driven synthesis of protein NS1 in parvoviral DNA amplification and gene expression, the stringent S-phase dependency of promoter P4 is likely a major determinant of the absolute requirement of the minute virus of mice for host cell proliferation. PMID- 9418889 TI - A hydrophobic segment within the 81-amino-acid domain of TFIIIA from Saccharomyces cerevisiae is essential for its transcription factor activity. AB - Transcription factor IIIA (TFIIIA) binds to the internal control region of the 5S RNA gene as the first step in the in vitro assembly of a TFIIIB-TFIIIC-TFIIIA-DNA transcription complex. An 81-amino-acid domain that is present between zinc fingers 8 and 9 of TFIIIA from Saccharomyces cerevisiae is essential for the transcription factor activity of this protein (C. A. Milne and J. Segall, J. Biol. Chem. 268:11364-11371, 1993). We have monitored the effect of mutations within this domain on the ability of TFIIIA to support transcription of the 5S RNA gene in vitro and to maintain cell viability. TFIIIA with internal deletions that removed residues 282 to 315, 316 to 334, 328 to 341, or 342 to 351 of the 81 amino-acid domain retained activity, whereas TFIIIA with a deletion of the short leucine-rich segment 352NGLNLLLN359 at the carboxyl-terminal end of this domain was devoid of activity. Analysis of the effects of double and quadruple mutations in the region extending from residue 336 to 364 confirmed that hydrophobic residues in this portion of the 81-amino-acid domain, particularly L343, L347, L354, L356, L357, and L358, and to a lesser extent F336 and L337, contributed to the ability of TFIIIA to promote transcription. We propose that these hydrophobic residues play a role in mediating an interaction between TFIIIA and another component of the transcriptional machinery. We also found that TFIIIA remained active if either zinc finger 8 or zinc finger 9 was disrupted by mutation but that TFIIIA containing a disruption of both zinc finger 8 and zinc finger 9 was inactive. PMID- 9418890 TI - Cln3-associated kinase activity in Saccharomyces cerevisiae is regulated by the mating factor pathway. AB - The Saccharomyces cerevisiae cell cycle is arrested in G1 phase by the mating factor pathway. Genetic evidence has suggested that the G1 cyclins Cln1, Cln2, and Cln3 are targets of this pathway whose inhibition results in G1 arrest. Inhibition of Cln1- and Cln2-associated kinase activity by the mating factor pathway acting through Far1 has been described. Here we report that Cln3 associated kinase activity is inhibited by mating factor treatment, with dose response and timing consistent with involvement in cell cycle arrest. No regulation of Cln3-associated kinase was observed in a fus3 kss1 strain deficient in mating factor pathway mitogen-activated protein (MAP) kinases. Inhibition occurs mainly at the level of specific activity of Cln3-Cdc28 complexes. Inhibition of the C-terminally truncated Cln3-1-associated kinase is not observed; such truncations were previously identified genetically as causing resistance to mating factor-induced cell cycle arrest. Regulation of Cln3 associated kinase specific activity by mating factor treatment requires Far1. Overexpression of Far1 restores inhibition of C-terminally truncated Cln3-1 associated kinase activity. G2/M-arrested cells are unable to regulate Cln3 associated kinase, possibly because of cell cycle regulation of Far1 abundance. Inhibition of Cln3-associated kinase activity by the mating factor pathway may allow this pathway to block the earliest step in normal cell cycle initiation, since Cln3 functions as the most upstream G1-acting cyclin, activating transcription of the G1 cyclins CLN1 and CLN2 as well as of the S-phase cyclins CLB5 and CLB6. PMID- 9418891 TI - Isolation of estrogen-responsive genes with a CpG island library. AB - In order to isolate novel estrogen-responsive genes, we utilized a CpG island library in which the regulatory regions of genes are enriched. CpG islands were screened for the ability to bind to a recombinant estrogen receptor protein with a genomic binding site (GBS) cloning method. Six CpG islands were selected, and they contained perfect, imperfect, and/or multiple half-palindromic estrogen responsive elements (EREs). Northern blot analysis of various human cells showed that all these genomic fragments hybridized to specific mRNAs, suggesting that the genes associated with these EREs might be transcribed in human cells. Then cDNAs associated with two of them, EB1 and EB9, were isolated from libraries of human placenta and MCF-7 cells derived from a human breast cancer, respectively. Both transcripts were increased by estrogen in MCF-7 cells. The increase is inhibited by actinomycin D but not by cycloheximide, indicating that no protein synthesis is required for the up-regulation. The cDNA associated with EB1 encodes a 114-amino-acid protein similar to the cytochrome c oxidase subunit VIIa, named COX7RP (cytochrome c oxidase subunit VII-related protein). The cDNA associated with EB9 is homologous only to an express sequence tag and was named EBAG9 (estrogen receptor-binding fragment-associated gene 9). The palindromic ERE of EB1 is located in an intron of COX7RP, and that of EB9 is in the 5' upstream region of the cDNA. Both EREs had significant estrogen-dependent enhancer activities in a chloramphenicol acetyltransferase assay, when they were inserted into the 5' upstream region of the chicken beta-globin promoter. We therefore propose that the CpG-GBS method described here for isolation of the DNA binding site from the CpG island library would be useful for identification of novel target genes of certain transcription factors. PMID- 9418892 TI - Regulation of sex-specific selection of fruitless 5' splice sites by transformer and transformer-2. AB - In Drosophila melanogaster, the fruitless (fru) gene controls essentially all aspects of male courtship behavior. It does this through sex-specific alternative splicing of the fru pre-mRNA, leading to the production of male-specific fru mRNAs capable of expressing male-specific fru proteins. Sex-specific fru splicing involves the choice between alternative 5' splice sites, one used exclusively in males and the other used only in females. Here we report that the Drosophila sex determination genes transformer (tra) and transformer-2 (tra-2) switch fru splicing from the male-specific pattern to the female-specific pattern through activation of the female-specific fru 5' splice site. Activation of female specific fru splicing requires cis-acting tra and tra-2 repeat elements that are part of an exonic splicing enhancer located immediately upstream of the female specific fru 5' splice site and are recognized by the TRA and TRA-2 proteins in vitro. This fru splicing enhancer is sufficient to promote the activation by tra and tra-2 of both a 5' splice site and the female-specific doublesex (dsx) 3' splice site, suggesting that the mechanisms of 5' splice site activation and 3' splice site activation may be similar. PMID- 9418893 TI - Adenovirus E1A-regulated transcription factor p120E4F inhibits cell growth and induces the stabilization of the cdk inhibitor p21WAF1. AB - Adenovirus E1A proteins influence cell growth and phenotype through physical interactions with cellular proteins that regulate basic processes such as cell cycle progression, DNA synthesis, and differentiation. p120E4F is a low-abundance cellular transcription factor that represses the adenovirus E4 promoter and is regulated by E1A, through a phosphorylation-induced reduction of its DNA binding activity, to permit activation of the E4 promoter during early infection. To determine the normal biological role of p120E4F, we assessed its ability to influence fibroblast cell growth and transformation. p120E4F suppressed NIH 3T3 fibroblast colony formation but had little effect when coexpressed with E1A and/or activated ras. Cells that overexpressed p120E4F were inhibited in their ability to enter S phase, had elevated levels of the cdk inhibitor p21WAF1, and reduced cyclin D-cdk4/6 kinase activity. The increase of p21WAF1 levels occurred through a p53-independent posttranscriptional mechanism that included a three- to fourfold increase in the half-life of p21WAF1 protein. Coexpression of activated ras with p120E4F stimulated cyclin D1 expression, elevated cyclin D-cdk4/6 kinase activity, and accelerated cell growth. These data suggest an important role for p120E4F in normal cell division and demonstrate that p21WAF1 can be regulated by protein turnover. PMID- 9418894 TI - A subunit of the anaphase-promoting complex is a centromere-associated protein in mammalian cells. AB - Sister chromatids in early mitotic cells are held together mainly by interactions between centromeres. The separation of sister chromatids at the transition between the metaphase and the anaphase stages of mitosis depends on the anaphase promoting complex (APC), a 20S ubiquitin-ligase complex that targets proteins for destruction. A subunit of the APC, called APC-alpha in Xenopus (and whose homologs are APC-1, Cut4, BIME, and Tsg24), has recently been identified and shown to be required for entry into anaphase. We now show that the mammalian APC alpha homolog, Tsg24, is a centromere-associated protein. While this protein is detected only during the prophase to the anaphase stages of mitosis in Chinese hamster cells, it is constitutively associated with the centromeres in murine cells. We show that there are two forms of this protein in mammalian cells, a soluble form associated with other components of the APC and a centromere-bound form. We also show that both the Tsg24 protein and the Cdc27 protein, another APC component, are bound to isolated mitotic chromosomes. These results therefore support a model in which the APC by ubiquitination of a centromere protein regulates the sister chromatid separation process. PMID- 9418895 TI - Expression of constitutively active IkappaB beta in T cells of transgenic mice: persistent NF-kappaB activity is required for T-cell immune responses. AB - The transcription factor NF-kappaB is normally sequestered in the cytoplasm by members of the IkappaB family, including IkappaB alpha, IkappaB beta, and the recently cloned IkappaB epsilon. Upon cellular activation, these inhibitors are rapidly phosphorylated on two amino-terminal serines, ubiquitinated, and degraded by the 26S proteasome, releasing a functional NF-kappaB. To determine the importance of IkappaB beta in NF-kappaB regulation in T cells, we generated transgenic mice expressing a constitutively active IkappaB beta mutant (mIkappaB beta) under the control of the lck promoter. The transgene contains the two critical N-terminal serine residues mutated to alanines and therefore no longer susceptible to degradation upon cell activation. mIkappaB beta is unable to totally displace IkappaB alpha from RelA-containing complexes, thus allowing a transient activation of NF-kappaB upon T-cell stimulation. However, mIkappaB beta completely blocks NF-kappaB activity after IkappaB alpha degradation. In addition, as a consequence of this inhibition, ikba expression is down regulated, along with that of other NF-kappaB-regulated genes. These transgenic mice have a significant reduction in the peripheral T-cell population, especially CD8+ cells. The remaining T cells have impaired proliferation in response to phorbol 12 myristate 13-acetate plus phytohemagglutinin or calcium ionophore but not to anti CD3/anti-CD28 costimulation. As a result of these alterations, transgenic animals present defects in immune responses such as delayed-type hypersensitivity and the generation of specific antibodies against T-cell-dependent antigens. These results show that in nonstimulated T cells, IkappaB beta cannot efficiently displace IkappaB alpha bound to RelA-containing complexes and that persistent NF kappaB activity is required for proper T-cell responses in vivo. PMID- 9418896 TI - cDNA cloning and characterization of the human U3 small nucleolar ribonucleoprotein complex-associated 55-kilodalton protein. AB - The eukaryotic nucleolus contains a large number of small RNA molecules (snoRNAs) which, in the form of small nucleolar ribonucleoprotein complexes (snoRNPs), are involved in the processing and modification of pre-rRNA. The most abundant and one of the best-conserved snoRNAs is the U3 RNA. So far, only one human U3 snoRNA associated protein, fibrillarin, has been characterized. Previously, the U3 snoRNPwas purified from CHO cells, and three proteins of 15, 50, and 55 kDa were found to copurify with the U3 snoRNA (B. Lubben, C. Marshallsay, N. Rottmann, and R. Luhrmann, Nucleic Acids Res. 21:5377-5385, 1993). Here we report the cDNA cloning and characterization of the human U3 snoRNP-associated 55-kDa protein. The isolated cDNA codes for a novel nucleolar protein which is specifically associated with the U3 snoRNA. This protein, referred to as hU3-55k, is the first characterized U3 snoRNP-specific protein from humans. hU3-55k is a new member of the family of WD-40 repeat proteins and is conserved throughout evolution. It appears that the C-terminal end of hU3-55k is required for nucleolar localization and U3 snoRNA binding. PMID- 9418897 TI - The C-terminal domain of B-Myb acts as a positive regulator of transcription and modulates its biological functions. AB - The myb gene family consists of three members, named A-, B-, and c-myb. All three members of this family encode nuclear proteins that bind DNA in a sequence specific manner and function as regulators of transcription. In this report, we have examined the biochemical and biological activities of murine B-myb and compared these properties with those of murine c-myb. In transient transactivation assays, murine B-myb exhibited transactivation potential comparable to that of c-myb. An analysis of deletion mutants of B-myb and c-myb showed that while the C-terminal domain of c-Myb acts as a negative regulator of transcriptional transactivation, the C-terminal domain of B-Myb functions as a positive enhancer of transactivation. To compare the biological activities of c myb and B-myb, the two genes were overexpressed in 32Dcl3 cells, which are known to undergo terminal differentiation into granulocytes in the presence of granulocyte colony-stimulating factor (G-CSF). We observed that c-myb blocked the G-CSF-induced terminal differentiation of 32Dcl3 cells, resulting in their continued proliferation in the presence of G-CSF. In contrast, ectopic overexpression of B-myb blocked the ability of 32D cells to proliferate in the presence of G-CSF and accelerated the G-CSF-induced granulocytic differentiation of these cells. Similar studies with B-myb-c-myb chimeras showed that only chimeras that contained the C-terminal domain of B-Myb were able to accelerate the G-CSF-induced terminal differentiation of 32Dcl3 cells. These studies show that c-myb and B-myb do not exhibit identical biological activities and that the carboxyl-terminal regulatory domain of B-Myb plays a critical role in its biological function. PMID- 9418898 TI - Nab1, a corepressor of NGFI-A (Egr-1), contains an active transcriptional repression domain. AB - Nab proteins constitute an evolutionarily conserved family of corepressors that specifically interact with and repress transcription mediated by three members of the NGFI-A (Egr-1, Krox24, zif/268) family of immediate-early gene transcription factors, which includes NGFI-C, Krox20, and Egr3. We explored the mechanism of Nab1 repression and identified structural domains required for Nab1 function. Nab1 does not act by blocking DNA binding or nuclear localization of NGFI-A. In fact, Nab1 repression is not unique to NGFI-A because multiple types of non-NGFI A activation domains were repressed, as was a heterologous transcription factor carrying the NGFI-A R1 domain, which is required for Nab1 interaction. Additionally, Nab1 tethered directly to DNA repressed constitutively active promoters. Tethered repression was not dependent on the identity of the basal promoter elements, the presence of a distal enhancer, or the distance separating the binding sites from the promoter. These results suggest that Nab1 repression is not specific to particular activators and that Nab1 is an active repressor that works by a direct mechanism. We identified a bipartite-like nuclear localization sequence and localized the repression function to the Nab conserved domain 2 (NCD2), a region found in the carboxy-terminal half of all Nab proteins. Three small regions of homology between Nab1 and previously characterized corepressors, Dr1 and E1b 55-kDa protein, were identified within NCD2. Replacement mutagenesis of residues conserved between these proteins interfered with Nab1 repression, although Nab1 does not function by the same mechanism as Dr1. The human NAB1 genomic locus was mapped to chromosome 2q32.3-33. PMID- 9418900 TI - MYC abrogates p53-mediated cell cycle arrest in N-(phosphonacetyl)-L-aspartate treated cells, permitting CAD gene amplification. AB - Genomic instability, including the ability to undergo gene amplification, is a hallmark of neoplastic cells. Similar to normal cells, "nonpermissive" REF52 cells do not develop resistance to N-(phosphonacetyl)-L-aspartate (PALA), an inhibitor of the synthesis of pyrimidine nucleotides, through amplification of cad, the target gene, but instead undergo protective, long-term, p53-dependent cell cycle arrest. Expression of exogenous MYC prevents this arrest and allows REF52 cells to proceed to mitosis when pyrimidine nucleotides are limiting. This results in DNA breaks, leading to cell death and, rarely, to cad gene amplification and PALA resistance. Pretreatment of REF52 cells with a low concentration of PALA, which slows DNA replication but does not trigger cell cycle arrest, followed by exposure to a high, selective concentration of PALA, promotes the formation of PALA-resistant cells in which the physically linked cad and endogenous N-myc genes are coamplified. The activated expression of endogenous N-myc in these pretreated PALA-resistant cells allows them to bypass the p53-mediated arrest that is characteristic of untreated REF52 cells. Our data demonstrate that two distinct events are required to form PALA-resistant REF52 cells: amplification of cad, whose product overcomes the action of the drug, and increased expression of N-myc, whose product overcomes the PALA-induced cell cycle block. These paired events occur at a detectable frequency only when the genes are physically linked, as cad and N-myc are. In untreated REF52 cells overexpressing N-MYC, the level of p53 is significantly elevated but there is no induction of p21waf1 expression or growth arrest. However, after DNA is damaged, the activated p53 executes rapid apoptosis in these REF52/N-myc cells instead of the long-term protective arrest seen in REF52 cells. The predominantly cytoplasmic localization of stabilized p53 in REF52/N-myc cells suggests that cytoplasmic retention may help to inactivate the growth-suppressing function of p53. PMID- 9418899 TI - Constitutive activation of the aromatic hydrocarbon receptor. AB - The ligand-activated aromatic hydrocarbon receptor (AHR) dimerizes with the AHR nuclear translocator (ARNT) to form a functional complex that transactivates expression of the cytochrome P-450 CYP1A1 gene and other genes in the dioxin inducible [Ah] gene battery. Previous work from this laboratory has shown that the activity of the CYP1A1 enzyme negatively regulates this process. To study the relationship between CYP1A1 activity and Ah receptor activation we used CYP1A1 deficient mouse hepatoma c37 cells and CYP1A1- and AHR-deficient African green monkey kidney CV-1 cells. Using gel mobility shift and luciferase reporter gene expression assays, we found that c37 cells that had not been exposed to exogenous Ah receptor ligands already contained transcriptionally active AHR-ARNT complexes, a finding that we also observed in wild-type Hepa-1 cells treated with Ellipticine, a CYP1A1 inhibitor. In CV-1 cells, transient expression of AHR and ARNT leads to high levels of AHR-ARNT-dependent luciferase gene expression even in the absence of an agonist. Using a green fluorescent protein-tagged AHR, we showed that elevated reporter gene expression correlates with constitutive nuclear localization of the AHR. Transcriptional activation of the luciferase reporter gene observed in CV-1 cells is significantly decreased by (i) expression of a functional CYP1A1 enzyme, (ii) competition with chimeric or truncated AHR proteins containing the AHR ligand-binding domain, and (iii) treatment with the AHR antagonist alpha-naphthoflavone. These results suggest that a CYP1A1 substrate, which accumulates in cells lacking CYP1A1 enzymatic activity, is an AHR ligand responsible for endogenous activation of the Ah receptor. PMID- 9418902 TI - 4-1BB and Ox40 are members of a tumor necrosis factor (TNF)-nerve growth factor receptor subfamily that bind TNF receptor-associated factors and activate nuclear factor kappaB. AB - Members of the tumor necrosis factor (TNF)-nerve growth factor (NGF) receptor family have been shown to be important costimulatory molecules for cellular activation. 4-1BB and Ox40 are two recently described members of this protein family which are expressed primarily on activated T cells. To gain insight into the signaling pathways employed by these factors, yeast two-hybrid library screens were performed with the cytoplasmic domains of 4-1BB and Ox40 as baits. TNF receptor-associated factor 2 (TRAF2) was identified as an interacting protein in both screens. The ability of both 4-1BB and Ox40 to interact with TRAF2 was confirmed in mammalian cells by coimmunoprecipitation studies. When the binding of the receptors to other TRAF proteins was investigated, 4-1BB and Ox40 displayed distinct binding patterns. While 4-1BB bound TRAF2 and TRAF1, Ox40 interacted with TRAF3 and TRAF2. Using deletion and alanine scanning analysis, we defined the elements in the cytoplasmic domains of both receptors that mediate these interactions. The 4-1BB receptor was found to have two independent stretches of acidic residues that can mediate association of the TRAF molecules. In contrast, a single TRAF binding domain was identified in the cytoplasmic tail of Ox40. The cytoplasmic domains of both receptors were shown to activate nuclear factor kappaB in a TRAF-dependent manner. Taken together, our results indicate that 4-1BB and Ox40 bind TRAF proteins to initiate a signaling cascade leading to activation of nuclear factor kappaB. PMID- 9418901 TI - Nuclear accumulation of p21Cip1 at the onset of mitosis: a role at the G2/M-phase transition. AB - Cell cycle arrest in G1 in response to ionizing radiation or senescence is believed to be provoked by inactivation of G1 cyclin-cyclin-dependent kinases (Cdks) by the Cdk inhibitor p21(Cip1/Waf1/Sdi1). We provide evidence that in addition to exerting negative control of the G1/S phase transition, p21 may play a role at the onset of mitosis. In nontransformed fibroblasts, p21 transiently reaccumulates in the nucleus near the G2/M-phase boundary, concomitant with cyclin B1 nuclear translocation, and associates with a fraction of cyclin A-Cdk and cyclin B1-Cdk complexes. Premitotic nuclear accumulation of cyclin B1 is not detectable in cells with low p21 levels, such as fibroblasts expressing the viral human papillomavirus type 16 E6 oncoprotein, which functionally inactivates p53, or in tumor-derived cells. Moreover, synchronized E6-expressing fibroblasts show accelerated entry into mitosis compared to wild-type cells and exhibit higher cyclin A- and cyclin B1-associated kinase activities. Finally, primary embryonic fibroblasts derived from p21-/- mice have significantly reduced numbers of premitotic cells with nuclear cyclin B1. These data suggest that p21 promotes a transient pause late in G2 that may contribute to the implementation of late cell cycle checkpoint controls. PMID- 9418903 TI - A role for the putative tumor suppressor Bin1 in muscle cell differentiation. AB - Bin1 is a Myc-interacting protein with features of a tumor suppressor. The high level of Bin1 expression in skeletal muscle prompted us to investigate its role in muscle differentiation. Significant levels of Bin1 were observed in undifferentiated C2C12 myoblasts, a murine in vitro model system. Induction of differentiation by growth factor withdrawal led to an upregulation of Bin1 mRNA and to the generation of higher-molecular-weight forms of Bin1 protein by alternate splicing. While Bin1 in undifferentiated cells was localized exclusively in the nucleus, differentiation-associated isoforms of Bin1 were found in the cytoplasm as well. To examine the function of Bin1 during differentiation, we generated stable cell lines that express exogenous human Bin1 cDNA in the sense or antisense orientation. Cells overexpressing Bin1 grew more slowly than control cells and differentiated more rapidly when deprived of growth factors. In contrast, C2C12 cells expressing antisense Bin1 showed an impaired ability to undergo differentiation. Taken together, the results indicated that Bin1 expression, structure, and localization are tightly regulated during muscle differentiation and suggested that Bin1 plays a functional role in the differentiation process. PMID- 9418904 TI - Role of UEV-1, an inactive variant of the E2 ubiquitin-conjugating enzymes, in in vitro differentiation and cell cycle behavior of HT-29-M6 intestinal mucosecretory cells. AB - By means of differential RNA display, we have isolated a cDNA corresponding to transcripts that are down-regulated upon differentiation of the goblet cell-like HT-29-M6 human colon carcinoma cell line. These transcripts encode proteins originally identified as CROC-1 on the basis of their capacity to activate transcription of c-fos. We show that these proteins are similar in sequence, and in predicted secondary and tertiary structure, to the ubiquitin-conjugating enzymes, also known as E2. Despite the similarities, these proteins lack a critical cysteine residue essential for the catalytic activity of E2 enzymes and, in vitro, they do not conjugate or transfer ubiquitin to protein substrates. These proteins constitute a distinct subfamily within the E2 protein family and are highly conserved in phylogeny from yeasts to mammals. Therefore, we have designated them UEV (ubiquitin-conjugating E2 enzyme variant) proteins, defined as proteins similar in sequence and structure to the E2 ubiquitin-conjugating enzymes but lacking their enzymatic activity (HW/GDB-approved gene symbol, UBE2V). At least two human genes code for UEV proteins, and one of them, located on chromosome 20q13.2, is expressed as at least four isoforms, generated by alternative splicing. All human cell types analyzed expressed at least one of these isoforms. Constitutive expression of exogenous human UEV in HT-29-M6 cells inhibited their capacity to differentiate upon confluence and caused both the entry of a larger proportion of cells in the division cycle and an accumulation in G2-M. This was accompanied with a profound inhibition of the mitotic kinase, cdk1. These results suggest that UEV proteins are involved in the control of differentiation and could exert their effects by altering cell cycle distribution. PMID- 9418905 TI - Requirement for phospholipase C-gamma1 enzymatic activity in growth factor induced mitogenesis. AB - The cytoplasmic regions of the receptors for epidermal growth factor (EGF) and platelet-derived growth factor (PDGF) bind and activate phospholipase C-gamma1 (PLC-gamma1) and other signaling proteins in response to ligand binding outside the cell. Receptor binding by PLC-gamma1 is a function of its SH2 domains and is required for growth factor-induced cell cycle progression into the S phase. Microinjection into MDCK epithelial cells and NIH 3T3 fibroblasts of a polypeptide corresponding to the noncatalytic SH2-SH2-SH3 domains of PLC-gamma1 (PLC-gamma1 SH2-SH2-SH3) blocked growth factor-induced S-phase entry. Treatment of cells with diacylglycerol (DAG) or DAG and microinjected inositol-1,4,5 triphosphate (IP3), the products of activated PLC-gamma1, did not stimulate cellular DNA synthesis by themselves but did suppress the inhibitory effects of the PLC-gamma1 SH2-SH2-SH3 polypeptide but not the cell cycle block imposed by inhibition of the adapter protein Grb2 or p21 Ras. Two c-fos serum response element (SRE)-chloramphenicol acetyltransferase (CAT) reporter plasmids, a wild type version, wtSRE-CAT, and a mutant, pm18, were used to investigate the function of PLC-gamma1 in EGF- and PDGF-induced mitogenesis. wtSRE-CAT responds to both protein kinase C (PKC)-dependent and -independent signals, while the mutant, pm18, responds only to PKC-independent signals. Microinjection of the dominant-negative PLC-gamma1 SH2-SH2-SH3 polypeptide greatly reduced the responses of wtSRE-CAT to EGF stimulation in MDCK cells and to PDGF stimulation in NIH 3T3 cells but had no effect on the responses of mutant pm18. These results indicate that in addition to Grb2-mediated activation of Ras, PLC-gamma1-mediated DAG production is required for EGF- and PDGF-induced S-phase entry and gene expression, possibly through activation of PKC. PMID- 9418906 TI - Numb-associated kinase interacts with the phosphotyrosine binding domain of Numb and antagonizes the function of Numb in vivo. AB - During asymmetric cell division, the membrane-associated Numb protein localizes to a crescent in the mitotic progenitor and is segregated predominantly to one of the two daughter cells. We have identified a putative serine/threonine kinase, Numb-associated kinase (Nak), which interacts physically with the phosphotyrosine binding (PTB) domain of Numb. The PTB domains of Shc and insulin receptor substrate bind to an NPXY motif which is not present in the region of Nak that interacts with Numb PTB domain. We found that the Numb PTB domain but not the Shc PTB domain interacts with Nak through a peptide of 11 amino acids, implicating a novel and specific protein-protein interaction. Overexpression of Nak in the sensory organs causes both daughters of a normally asymmetric cell division to adopt the same cell fate, a transformation similar to the loss of numb function phenotype and opposite the cell fate transformation caused by overexpression of Numb. The frequency of cell fate transformation is sensitive to the numb gene dosage, as expected from the physical interaction between Nak and Numb. These findings indicate that Nak may play a role in cell fate determination during asymmetric cell divisions. PMID- 9418907 TI - Human IAP-like protein regulates programmed cell death downstream of Bcl-xL and cytochrome c. AB - The gene encoding human IAP-like protein (hILP) is one of several mammalian genes with sequence homology to the baculovirus inhibitor-of-apoptosis protein (iap) genes. Here we show that hILP can block apoptosis induced by a variety of extracellular stimuli, including UV light, chemotoxic drugs, and activation of the tumor necrosis factor and Fas receptors. hILP also protected against cell death induced by members of the caspase family, cysteine proteases which are thought to be the principal effectors of apoptosis. hILP and Bcl-xL were compared for their ability to affect several steps in the apoptotic pathway. Redistribution of cytochrome c from mitochondria, an early event in apoptosis, was not blocked by overexpression of hILP but was inhibited by Bcl-xL. In contrast, hILP, but not Bcl-xL, inhibited apoptosis induced by microinjection of cytochrome c. These data suggest that while Bcl-xL may control mitochondrial integrity, hILP can function downstream of mitochondrial events to inhibit apoptosis. PMID- 9418908 TI - Pex19p, a farnesylated protein essential for peroxisome biogenesis. AB - We report the identification and molecular characterization of Pex19p, an oleic acid-inducible, farnesylated protein of 39.7 kDa that is essential for peroxisome biogenesis in Saccharomyces cerevisiae. Cells lacking Pex19p are characterized by the absence of morphologically detectable peroxisomes and mislocalization of peroxisomal matrix proteins to the cytosol. The human HK33 gene product was identified as the putative human ortholog of Pex19p. Evidence is provided that farnesylation of Pex19p takes place at the cysteine of the C-terminal CKQQ amino acid sequence. Farnesylation of Pex19p was shown to be essential for the proper function of the protein in peroxisome biogenesis. Pex19p was shown to interact with Pex3p in vivo, and this interaction required farnesylation of Pex19p. PMID- 9418909 TI - Effects of p21(Cip1/Waf1) at both the G1/S and the G2/M cell cycle transitions: pRb is a critical determinant in blocking DNA replication and in preventing endoreduplication. AB - It has been proposed that the functions of the cyclin-dependent kinase inhibitors p21(Cip1/Waf1) and p27Kip1 are limited to cell cycle control at the G1/S-phase transition and in the maintenance of cellular quiescence. To test the validity of this hypothesis, p21 was expressed in a diverse panel of cell lines, thus isolating the effects of p21 activity from the pleiotropic effects of upstream signaling pathways that normally induce p21 expression. The data show that at physiological levels of accumulation, p21, in addition to its role in negatively regulating the G1/S transition, contributes to regulation of the G2/M transition. Both G1- and G2-arrested cells were observed in all cell types, with different preponderances. Preponderant G1 arrest in response to p21 expression correlated with the presence of functional pRb. G2 arrest was more prominent in pRb-negative cells. The arrest distribution did not correlate with the p53 status, and proliferating-cell nuclear antigen (PCNA) binding activity of p21 did not appear to be involved, since p27, which lacks a PCNA binding domain, produced similar arrest distributions [corrected], DNA endoreduplication occurred in pRb-negative but not in pRb-positive cells, suggesting that functional pRb is necessary to prevent DNA replication in p21 G2-arrested cells. These results suggest that the primary target of the Cip/Kip family of inhibitors leading to efficient G1 arrest as well as to blockade of DNA replication from either G1 or G2 phase is the pRb regulatory system. Finally, the tendency of Rb-negative cells to undergo endoreduplication cycles when p21 is expressed may have negative implications in the therapy of Rb-negative cancers with genotoxic agents that activate the p53/p21 pathway. PMID- 9418910 TI - LIM protein KyoT2 negatively regulates transcription by association with the RBP J DNA-binding protein. AB - The RBP-J/Su(H) DNA-binding protein plays a key role in transcriptional regulation by targeting Epstein-Barr virus nuclear antigen 2 (EBNA2) and the intracellular portions of Notch receptors to specific promoters. Using the yeast two-hybrid system, we isolated a LIM-only protein, KyoT, which physically interacts with RBP-J. Differential splicing gave rise to two transcripts of the KyoT gene, KyoT1 and KyoT2, that encoded proteins with four and two LIM domains, respectively. With differential splicing resulting in deletion of an exon, KyoT2 lacked two LIM domains from the C terminus and had a frameshift in the last exon, creating the RBP-J-binding region in the C terminus. KyoT1 had a negligible level of interaction with RBP-J. Strong expression of KyoT mRNAs was detected in skeletal muscle and lung, with a predominance of KyoT1 mRNA. When expressed in F9 embryonal carcinoma cells, KyoT1 and KyoT2 were localized in the cytoplasm and the nucleus, respectively. The binding site of KyoT2 on RBP-J overlaps those of EBNA2 and Notchl but is distinct from that of Hairless, the negative regulator of RBP-J-mediated transcription in Drosophila. KyoT2 but not KyoT1 repressed the RBP J-mediated transcriptional activation by EBNA2 and Notch1 by competing with them for binding to RBP-J and by dislocating RBP-J from DNA. KyoT2 is a novel negative regulatory molecule for RBP-J-mediated transcription in mammalian systems. PMID- 9418913 TI - Is it necessary to suture the platysma muscles on the midline to improve the cervical profile? An anatomic study using 20 cadavers. AB - To ameliorate the cervicomental angle, most surgeons suggest different techniques of platysmaplasty. The aim of this anatomic study is to find a simple answer to the following question: Is suturing of the anterior edges of the platysma muscles during platysmaplasty the best procedure to use to obtain the best concave anterior neck angle? Three different surgical techniques using platysma muscle flaps were used on 20 cadavers prepared for anatomic dissection. Each piece of dissection was controlled by a radiograph of the profile of the cervical region before and after the application of these different techniques. Cephalometric measures were made and statistically analyzed. The analysis of the results demonstrates that the best concave anterior neck angle to perform platysmaplasty is one in which the platysma muscle flap is shifted posterosuperiorly but without suturing the medial borders of the platysma muscles. Suturing the midline does not deepen the concavity in the front of the neck. PMID- 9418912 TI - Plastic surgical considerations in lighting injuries. AB - Lightning injuries affect 1,000 to 1,500 people per year in the United States. While fatalities are uncommon, lightning frequently causes injuries within the treatment purview of the plastic surgeon. Recognition of common patterns of lightning injury, their prognosis, and treatments are important for the plastic surgeon. Lightning injury, although electrical, is very different from other more common electrical injuries. Burn injuries are seen, but are usually superficial and heal without surgery. Deep-tissue injury is rare. Neurovascular compromise of the extremities is common and may lead to plastic surgical consultation for intervention. Spontaneous recovery is the rule, however. Several patients are described who illustrate these injuries, and treatment guidelines are proposed. PMID- 9418911 TI - Footprint analysis of the RAG protein recombination signal sequence complex for V(D)J type recombination. AB - We have studied the interaction between recombination signal sequences (RSSs) and protein products of the truncated forms of recombination-activating genes (RAG) by gel mobility shift, DNase I footprinting, and methylation interference assays. Methylation interference with dimethyl sulfate demonstrated that binding was blocked by methylation in the nonamer at the second-position G residue in the bottom strand and at the sixth- and seventh-position A residues in the top strand. DNase I footprinting experiments demonstrated that RAG1 alone, or even a RAG1 homeodomain peptide, gave footprint patterns very similar to those obtained with the RAG1-RAG2 complex. In the heptamer, partial methylation interference was observed at the sixth-position A residue in the bottom strand. In DNase I footprinting, the heptamer region was weakly protected in the bottom strand by RAG1. The effects of RSS mutations on RAG binding were evaluated by DNA footprinting. Comparison of the RAG-RSS footprint data with the published Hin model confirmed the notion that sequence-specific RSS-RAG interaction takes place primarily between the Hin domain of the RAG1 protein and adjacent major and minor grooves of the nonamer DNA. PMID- 9418914 TI - Endoscopic rectus harvest: a simplified sheath-saving technique. AB - The potential benefits of harvesting the rectus abdominis muscle endoscopically are great. To date no simple, reliable method has been developed for harvest based on the inferior epigastric artery and transferred to the living patient. We have devised a simplified endoscopically assisted method of harvest on a cadaveric model that adds speed and the advantage of a grossly intact sheath to other published methods. It has been used successfully in a clinical situation. Two-portal access and triplanar dissection are the key points in this method. PMID- 9418915 TI - Carbon dioxide laser ablation of anogenital condyloma acuminata in pediatric patients. AB - The treatment of anogenital condyloma acuminata in pediatric patients is difficult, with a wide range of treatment strategies that yield variable success. Treatment regimens must consider the patient age, and etiology, location, and severity of lesions. We report our experience using the carbon dioxide (CO2) laser to ablate these lesions. A retrospective review of a single surgeon's series of 17 consecutive patients was performed. A staging system was developed and used to document the extent of disease as related to prognosis, recurrence rates, and treatment options. There were 17 patients (5 males and 12 females). Eleven patients (65%) were treated after failing previous treatment. Perianal disease was noted in 14 of 17 patients (82%). No patients presented with stage I disease, 7 patients (41%) presented with stage II, 5 patients (29%) presented with stage III, and 5 patients (29%) presented with stage IV disease. Recurrence occurred in 4 of 17 patients (23%). Persistent disease occurred in 1 patient (6%). CO2 laser vaporization of anogenital condyloma acuminata in pediatric patients is safe, relatively atraumatic to the child, and associated with favorable recurrence rates. A staging scheme has been proposed in predicting recurrences, complications, and guiding therapy. PMID- 9418916 TI - A retrospective analysis of revision sphincter pharyngoplasty. AB - This retrospective study was undertaken to determine the revision rate for dynamic sphincter pharyngoplasty (DSP) at the University of Michigan Medical Center to analyze the determinants contributing to the need for revision pharyngoplasties, and ultimately to improve primary pharyngoplasty to avoid the need for revision. The records of 30 children with repaired palatal clefts who presented with velopharyngeal insufficiency and hypernasal speech, and who underwent DSP from January 1988 through July 1994 were reviewed. Clinical follow up ranged from 6 to 48 months (mean, 20.2 months). Seven of the original 30 patients (23%) had persistent, moderate-to-severe hypernasality that required reoperation, while 1 patient (3%) demonstrated hyponasality requiring revision. Seven of 8 patients who underwent revision pharyngoplasty had acceptable speech after revision. Dehiscences, low-lying pharyngoplasty flaps, and end-to-end suturing of the flaps were the main determinants resulting in the need for revision. In our study, female gender and older age was associated with a higher success of primary operation. PMID- 9418917 TI - Reversed arterial flow in free flap surgery for leg reconstruction. AB - A lower leg open fracture with an accompanying extensive soft-tissue defect may require free flap transfer to resurface the wound. In open-leg fracture injuries the major arteries are frequently damaged during the injurious event. When the antegrade arterial blood flow is not available for arterial inflow to the free flap, end-to-side arterial anastomoses or a venous graft from a proximal healthy artery is an appropriate solution. Reversed arterial flow is another technique that may be used to provide arterial inflow to the free flap. We report 5 patients with open tibial fracture injuries that were successfully covered using latissimus dorsi muscle free flaps with reverse arterial inflow and either antegrade venous outflow (4 patients) or retrograde outflow (1 patient). This technique is indicated when (1) the defect is not located at the site adjacent to the residual antegrade artery, (2) a long vein graft might pass through severely scarred soft tissue, or (3) after failure in end-to-side anastomosis with accompanying insufficient antegrade proximal arterial flow but good distal arterial reflux. PMID- 9418918 TI - Mapping of the human body skin with laser Doppler flowmetry. AB - The purpose of this study was to evaluate cutaneous blood flow in the human body to better establish normal flow ranges and to determine whether the location, side of the body, sex, and age affect the flow range at seven different sites. From March 1993 to February 1994 a Periflux system 4001 laser Doppler flowmeter was used to measure the cutaneous microflow circulation of 1,680 selected points in 120 volunteers. The mean normal cutaneous blood flow of 120 volunteers was between 4 and 9 perfusion units (PU) except in the head, neck, hand, and foot. Mean cutaneous blood flow measurements were as follows: the upper arm, 6.6 +/- 1.20 PU (mean +/- SE); the forearm, 6.7 +/- 1.95 PU; the thorax, 7.1 +/- 1.72 PU; the flank, 6.3 +/- 1.23 PU; the abdomen, 5.3 +/- 1.79 PU; the thigh, 4.8 +/- 1.34 PU; the lower leg, 4.6 +/- 1.39 PU (p < 0.05). The blood flow of the thorax was highest and was 54% higher than the lower leg. The group of teenagers had the highest cutaneous blood flow, with an average value of 6.9 +/- 0.62 PU. The group of subjects in their sixties proved to have the lowest cutaneous blood flow-32% less than the teenagers. Gender differences were not noted. There were no significant differences in blood flow in regard to the side of the body. PMID- 9418919 TI - Anatomy of the postauricular island "revolving door" flap ("flip-flop" flap). AB - Reconstruction following resection of auricular (cavum conchae) lesions may be done with a retroauricular rotation flap. Recently there has been revived interest in this elegant reconstructive procedure. Although the vascular anatomy of the area was studied, no direct study of flap anatomy was reported. Six fresh adult male cadaveric dissections of the retroauricular area were performed. The skin and underlying subcutaneous tissue layer were reflected to correspond with flap size, and anatomic structures were studied. Dissection was carried out on 12 ears. The origin of the occipital belly of the occipitofrontalis muscle arising from the posterior mastoid region was identified in four patients and only as part of the fascial layer overlying the posterior mastoid region. Only a small portion of the sternocleidomastoid tendon at best is possibly incorporated in the flap. It seems that only a negligible contribution to the flap is derived from the temporalis muscle. The posterior auricular muscle was identified in all patients and its origin from the skull was (in all patients) included or bordered the posterior flap region. The posterior auricular artery (PAA) was seen in all 12 dissections. The artery was adjacent to the styloid process medial to the parotid gland superficially between the auricular cartilage and the mastoid process. The PAA was then found on the periosteum of the mastoid process, ascending deep to the posterior auricular muscle. The flap seems to be a truly fasciocutaneous flap with small, questionable, superior and anteroinferior muscular contributions, and an inclusion of the rather small posterior auricular muscle. As reported in other studies, blood supply to the area seems to be derived from the PAA. PMID- 9418920 TI - Physiological roles of endothelium-derived nitric oxide in the epigastric island flaps of rabbits. AB - Nitric oxide (NO), identified as the mediator of endothelium-dependent relaxation of vascular smooth muscle, is known to cause a number of inflammatory conditions, especially in ischemia/reperfusion injury. This experimental study, using a rabbit epigastric island flap, was designed to investigate whether skin flap ischemia followed by reperfusion-influenced serum NO and c-GMP concentrations in the flap. In addition, we also investigated the premedicated effects of the NO synthase inhibitor and heparin on serum NO and c-GMP concentrations in skin flap ischemia/reperfusion. Serum NO concentration after 15, 30, 45, and 60 minutes of ischemia followed by reperfusion significantly increased compared with that in nonischemic control and elevated flaps. On the contrary, serum NO concentration was suppressed in L-NAME or aminoguanidine pretreated animals with ischemic group. Administration of heparin increased the serum NO concentration in elevated flaps, but suppressed it in ischemic flaps followed by reperfusion. The changes in serum c-GMP and NO concentrations were related in all of the experimental groups. These results suggest that NO may be derived from vascular endothelial cells and dilate peripheral vessels in compensation for ischemia. PMID- 9418921 TI - Reconstruction of the toddler diaphragm in severe anterolateral congenital diaphragmatic hernia with the reverse latissimus dorsi flap. AB - The management of infants with severe congenital diaphragmatic hernia (CDH) continues to evolve. When a prosthetic patch is placed in the neonatal period for pleuroperitoneal separation, it ultimately will require a subsequent reconstruction for progressive pulmonary or abdominal symptomatology. The reverse latissimus dorsi (RLD) flap has been used for reconstruction in only several reports in the last 12 years. In this paper, a patient with severe anterolateral CDH is reconstructed with the RLD flap on an elective basis at 2 years of age. Elective repair was performed for the particular indication of chest wall restriction imposed by the nonpliable Gore-Tex patch. In this case, use of the RLD flap alone without the use of synthetic mesh has resulted in satisfactory results with 17 months of follow up. PMID- 9418922 TI - Surgical treatment of solid facial edema: when everything else fails. AB - A case of persistent solid facial edema is presented to illustrate and discuss this rare, poorly understood condition. It is most frequently a sequela of acne vulgaris, but it can also be caused by other congenital, infectious, and inflammatory processes. This entity is notoriously difficult to treat. Many therapeutic modalities have been tried. Early treatments with isotretinoin, clofazimine, and ketotifen have been occasionally effective while no treatment has been proven successful in the later stages. This is probably the first time that surgical treatment of this problem has been reported. This experience and a review of the literature may prove helpful to other practitioners in our specialty. PMID- 9418923 TI - Infantile myofibromatosis: a solitary lesion involving the upper lip. AB - Infantile myofibromatosis is a rare disease characterized by myofibroblastic proliferation, and typically occurs in early infancy. There is a wide spectrum of clinical presentation, which may involve various kinds of tissues in the body. Skin and subcutaneous lesions were the types of tissue most often seen. Although a multicentric form may behave aggressively, a solitary form of the tumor is benign with the possibility of spontaneous regression. Conservative management is justified after proper pathological diagnosis if the tumor involves an aesthetically important area. A case of solitary infantile myofibromatosis involving the upper lip is presented. Partial excision for biopsy was performed and long-term observation was undertaken. The tumor disappeared 3 years postoperatively. PMID- 9418925 TI - Pedicled bone grafts to treat nasal bone defects associated with full-thickness loss of nose. AB - A periosteal pedicled bone graft from the maxillary buttress was used to reconstruct nasal bone loss with full-thickness dorsal nasal skin loss after a nasomaxillary skin graft inlay. The covering defect and lining defect were reconstructed using a forehead flap and a nasolabial flap. The vascularity of the bone flap was confirmed by bone scintigraphy on the seventh postoperative day. The results obtained were satisfactory and the framework was found to be stable during the follow-up period. PMID- 9418924 TI - Reconstruction of a short nose by full-thickness tissue transfer with osteotomy. AB - A severely shortened nose secondary to fracture of the middle third of the face was reconstructed by full-thickness tissue transfer with osteotomy. A nasalis musculocutaneous island flap, including the scar located in the nasal dorsum, was harvested for the lining. It was elevated based on the lateral nasal artery. The donor site defect of the flap and the covering defect were reconstructed with a forehead flap. The platform and framework of the nose were corrected with an en bloc osteotomy of the nasal bone, including the piriform aperture with a vascularized calvarial bone graft. Satisfactory and stable results were obtained in one stage along with the restoration of the relationships between the nose and the surrounding structures. PMID- 9418926 TI - Malignant fibrous histiocytoma degeneration in a patient with facial fibrous dysplasia. AB - Seventy-two patients with craniofacial fibrous dysplasia were treated during a period of 18 years. Only 1 patient (1.4%) developed malignant degeneration into malignant fibrous histiocytoma. Total excision including the gross mass and adjacent facial bone was performed at that time, however the patient refused an orbital exenteration and postoperative adjuvant therapy. Tumor recurrence developed 2 years later and the patient died. To our knowledge this is the first report describing malignant fibrous histiocytoma degeneration from a fibrous dysplasia localized to the facial bones. Although complete curative treatment could not be undertaken in this case, some conclusions can be drawn regarding the clinical course of this uncommon entity. PMID- 9418927 TI - Lupus vulgaris of the earlobe. AB - Cutaneous tuberculosis is rare today and is often confused with other granulomatous lesions. Its diagnosis remains bothersome, because detecting mycobacteria in skin lesions using a conventional laboratory examination remains difficult. A 59-year-old woman presented with lupus vulgaris of the earlobe. Surgical treatment was employed, with an initial diagnosis of hemangioma. After the operation the condition was diagnosed correctly and preventive antituberculous drugs were prescribed. The incidence of tuberculosis has increased since the mid-1980s chiefly due to the expansion of the HIV-infected, immunocompromised population. Thus the diagnosis and treatment of cutaneous tuberculosis has become once again important. PMID- 9418928 TI - Ulnar nerve compression at the wrist secondary to anomalous muscles: a patient with a variant of abductor digiti minimi. AB - Three patients with aberrant muscles passing through Guyon's canal are described. Each of the three patients experienced, among other symptoms, dysesthesias on the volar aspects of the little and ring fingers. The anomalous muscle in two of these patients arose from the antebrachial fascia just proximal to Guyon's canal, traveled through the canal, and inserted with the abductor digiti minimi muscle. This anomaly was bilateral in both patients. In the third patient the abductor digiti minimi muscle originated radially from the transverse carpal ligament, and this variant too was bilateral. Symptoms in each of the 3 patients resolved following surgery. Ulnar nerve compression at Guyon's canal is discussed and the literature is reviewed. Although anomalous muscles occur frequently in this location, the anatomic variant found in patient 3 has not yet been described in the literature. PMID- 9418929 TI - Rotation advancement in traumatic upper lip ectropion. AB - Rotation advancement is a well-known technique in cleft lip repair. This procedure corrects lip length and abnormal muscle insertion, and allows for accurate approximation of lip structures. It is a widely accepted standard of practice for congenital harelip repair that teaches fundamental principles of reconstructive surgery. An innovation is presented in which this procedure is used to correct a traumatic lip ectropion. The anatomic design is identical to a congenital cleft lip repair. The benefits of this approach are discussed in an interesting patient report. PMID- 9418930 TI - Giant cutaneous horn: a patient report. AB - A large cutaneous horn was excised from the left side of the nose and cheek of a 68-year-old woman. Reconstruction was performed with a split-thickness skin graft. Histologically the lesion represented squamous cell carcinoma. The nature of malignant degeneration in cutaneous horns is discussed. PMID- 9418931 TI - Malignant mixed tumor of the parotid with myoepithelial carcinomatous and papillary adenocarcinomatous components. AB - A patient with malignant mixed tumor of the parotid gland with myoepithelial carcinomatous and papillary cyst adenocarcinomatous components is presented. Clinically the patient had a giant mass in the parotid location for 20 years, with a growth progression during the last 6 months. The undefined connection of two carcinomatous components of malignant mixed tumor is reviewed. PMID- 9418932 TI - Recurrent giant cell tumor over a 23-year period. AB - Giant cell tumor recurrence is certainly not an uncommon event. In fact, recurrence rates as high as 50% have been reported. When recurrence does occur, it is usually within several years of the original tumor excision, if not sooner. The present report illustrates the recurrence of such a lesion twice over a 23 year period and serves to remind one that despite all attempts at complete excision, recurrence of giant cell tumor may still occur even decades following the initial resection. PMID- 9418933 TI - Two patients with penile keloids: a review of the literature. AB - Keloid formation on the penis is exceptionally rare even though the penis is frequently subjected to surgical manipulations such as circumcision. Review of the literature revealed a few reports of patients with penile keloids. A 13-year old Caucasian boy who had a pronounced penile keloid after circumcision and a 56 year-old Caucasian male who had keloid formation on the dorsum of the shaft of the penis after several attacks of penoscrotal hidradenitis suppurativa are presented. They comprise the fifth and the sixth cases of keloid formation in the literature to our knowledge. Intralesional steroid injection and surgical excision are the treatment modalities for this malady. Surgical excision appears to be the choice of treatment for larger lesions that are disabling. Recurrence is still possible, of course. PMID- 9418934 TI - A tale of two heartaches. PMID- 9418935 TI - Re: Augmentation mammoplasty associated with a severe systemic illness. PMID- 9418936 TI - Re: Bead and wire intermaxillary fixation. PMID- 9418937 TI - Osseous autografts covered with temporalis fascia: looking for greater survival. PMID- 9418938 TI - A patient with unusual vascular malformation of cheek to lip and deep tissues. PMID- 9418939 TI - Application of bilayer artificial skin to a defect following excisional biopsy for malignant melanomalike lesion. PMID- 9418940 TI - Sex differences in the enzymatic hydrolysis of acetylsalicylic acid by microsomes from various rat tissues. AB - We studied the in vitro hydrolysis of acetylsalicylic acid (ASA) to salicylic acid (SA) catalysed by microsomal preparations from liver, kidney, small intestine and stomach mucosas and blood serum of adult female and male rats. Hepatic microsomes from male rats had the highest specific activity: 42.3 +/- 6.0 nmol SA mg(-1) min(-1) (mean +/- SEM). Kidney, intestine, stomach and serum activities were 60, 30, 14 and 0.7% with regard to the liver. In contrast, gastric microsomes from female rats showed the highest specific activity: 53 +/- 22.1 nmol SA mg(-1) min(-1) (mean +/- SEM) whereas intestine, liver, kidney and serum activities were 60, 43, 40 and 1.7% with regard to the stomach mucosa. Hepatic, renal and intestinal microsomes had a pH optimum of 5-6. Male rats had Vmax and Km values of 95.5, 83.4 and 29.4 nmol SA mg(-1) min(-1) and 2.9, 1.27 and 6.4 mM, while for female rats they were 54.8, 75.8 and 59.4 nmol SA mg(-1) min(-1) and 2.6, 1.35 and 3.4 mM for hepatic, renal and intestinal microsomes, respectively. Parathion inhibited the hydrolysis of ASA with an IC50 of 1.2 x 10( 5) M for liver and kidney and 5 x 10(6) M for intestine from male rats. PMID- 9418941 TI - Micronuclei formation in bone marrow cells of rats treated with meothrin (synthetic pyrethroid). AB - The production of micronuclei in rat bone marrow cells by the synthetic pyrethroid insecticide meothrin was investigated. Three different doses of meothrin were orally administered to rats for 14 consecutive days. All tested doses of meothrin increased the frequency of micronucleated polychromatic erythrocytes but the increase was statistically significant only at the two highest doses. Meothrin also affected the rate of bone marrow cell proliferation, as determined by changes in the ratio of polychromatic erythrocytes to normochromatic erythrocytes. PMID- 9418942 TI - Trimellitic anhydride-sensitive mouse as an animal model for contact urticaria. AB - The respiratory allergen trimellitic anhydride (TMA) has been shown to induce IgE production and immediate ear swelling in mice sensitized to it. We studied whether TMA sensitivity could be used as an animal model for immunological contact urticaria. BALB/C mice were sensitized to TMA by topical applications. Groups of animals were pretreated on the ears with the glucocorticosteroid (GCS) betamethasone-17,21-dipropionate, the antihistamine (AH) diphenhydramine hydrochloride, the non-steroidal anti-inflammatory drug (NSAID) indomethacin or vehicle (VEH). Ears were challenged with TMA and ear thickness was measured at baseline and 1, 2, 4, 8 and 24 h after challenge. Trimellitic anhydride caused a significant biphasic ear swelling response with an early peak at 1-2 h, a plateau at 4 h and a late peak at 24 h. However, there was also an early swelling by TMA in non-sensitized mice, suggesting that non-immunological as well as immunological mechanisms contribute to early swelling by TMA. Glucocorticosteroid suppressed significantly the early and to some extent the late TMA responses, while AH suppressed only early and NSAID only late TMA responses. Ear swelling in TMA-sensitive BALB/C mice may represent a combination of immunological and non immunological contact urticaria and allergic contact dermatitis. Mice sensitive to TMA may be helpful in defining pharmacological agents affecting contact urticaria and the model is perhaps suitable for identification of some immunologically mediated contact urticants. PMID- 9418943 TI - Pharmacokinetics and material balance studies of diethylenetriamine trihydrochloride in the Fischer 344 rat following oral, endotracheal or intravenous dosing. AB - The metabolism and disposition of diethylenetriamine trihydrochloride (DETA.3HCl) were studied with regard to route of administration and dosage effects. Male Fischer 344 rats were administered 50 or 500 mg kg(-1) of [1,2-(14)C]-DETA.3HCl orally or endotracheally, and the fate of the 14C-radioactivity was followed for 48 h. The DETA.3HCl was readily absorbed from the gut or the lung, with bioavailabilities of 95% and 90%, respectively. It was distributed throughout the body, with the kidney attaining the highest concentration (about 2.5-5 times that of blood). The apparent volume of distribution determined from plasma concentration data following intravenous dosing (50 mg kg[-1]) was 486 ml kg(-1), consistent with distribution in the total body water. Urine and feces were the major routes of excretion, with only a small fraction eliminated as CO2. Excretion was quite rapid, with over 96% of the dose eliminated within 48 h. The principal component in the urine was unchanged DETA, suggesting that DETA.3HCl was not extensively metabolized. While the major metabolites has not been identified, there was evidence that DETA.3HCl was neither metabolized to ethylenediamine nor to the acid conjugates. There was indication that the metabolism of DETA.3HCl was saturated at 500 mg kg(-1), as there was a shift to a higher proportion of unchanged DETA excreted in the urine. There were no significant differences in material balance or pharmacokinetic parameters among animals receiving DETA.3HCl by the oral or endotracheal routes of administration. PMID- 9418944 TI - Studies on rat thyroid after oral administration of mancozeb: morphological and biochemical evaluations. AB - Mancozeb, an ethylenebisdithiocarbamate (EBDC), has been studied for its effects on rat thyroid. Single oral administration of mancozeb at different concentrations (9600, 12,000, 15,000 and 18,750 mg kg(-1) body wt) has derived the oral LD50 value as 15,000 mg kg(-1) body wt. in male rats. Mancozeb at repeated oral doses of 500, 1000 and 1500 mg kg(-1) day(-1) for periods of 30, 90, 180 and 360 days has produced dose-dependent signs of toxicity and death of animals. The fungicide caused a significant increase in thyroid/body weight ratio and histopathological changes. Reduced levels of thyroid radioiodine ([125]I) uptake, serum protein-bound iodine (PB[125]I), thyroxine (T4) and reduced activity of thyroid peroxidase (TPO) have also been observed after exposure to mancozeb. Thus, mancozeb has been shown to produce marked structural and functional changes in thyroid of rats. PMID- 9418945 TI - Investigations on immune parameters in welders. AB - The aim of the present investigation was to study the effects of welding fumes on the human immune system. Thirty male subjects who had regularly welded and 16 control persons without occupational exposure were examined. Cellular immunity was evaluated by phenotyping of peripheral leucocytes, measurement of mitogenic T cell response and T cell stimulation in a heterologous mixed lymphocyte reaction. Non-specific immune reactions were quantified by oxidative burst of granulocytes and monocytes and the cytotoxicity of lymphokine-activated killer (LAK) cells. Serum immunoglobulin levels and immunoglobulin production by stimulated B cells served to demonstrate humoral immune reactions. Welding fumes retarded the kinetics of DNA synthesis after phytohaemagglutinin stimulation of T cells and reduced the cytotoxic activity of LAK cells. No effects on lymphocytic subpopulations, mixed lymphocyte reaction, the phagocytosis of leucocytes or the production of immunoglobulins were observed. Several welders reported on recurrent respiratory infections or bronchitis, a few on allergic skin reactions and one worker was affected by asthmatic symptoms. With the exception of a reduced activity of LAK cells, these effects could not be related to any impairment of immune reactions as they were measured by the immunotoxicity tests applied. PMID- 9418946 TI - Glycyrrhizic acid inhibits arylamine N-acetyltransferase activity in Klebsiella pneumoniae in vitro. AB - Glycyrrhizic acid, one of the proposed chemopreventive drugs, was used to inhibit arylamine N-acetyltransferase (NAT) activity in Klebsiella pneumoniae, both in cytosol and intact bacteria. The NAT activity was measured by using high performance liquid chromatography to assay the amounts of 2-acetyl-aminofluorene and remaining 2-aminofluorene. The NAT activity in K. pneumoniae was inhibited by glycyrrhizic acid in a dose-dependent manner. The cytosol NAT activities were 0.675 +/- 0.028 nmol min(-1) mg(-1) protein for the acetylation of 2 aminofluorene without glycyrrhizic acid and 0.367 +/- 0.008 nmol min(-1) mg(-1) protein with 8 mM glycyrrhizic acid. The NAT activities measured from intact bacteria were 0.308 +/- 0.018 nmol min(-1) 10(-10) colony forming units for the acetylation of 2-aminofluorene without glycyrrhizic acid and 0.236 +/- 0.005 nmol min(-1) 10(-10) colony forming units in the presence of 8 mM glycyrrhizic acid. The inhibition of NAT activity by glycyrrhizic acid was demonstrated to remain for at least 4 h. The apparent Km and Vmax values calculated from cytosol NAT were 1.08 +/- 0.05 mM and 9.09 +/- 0.11 nmol min(-1) mg(-1) protein, respectively, for 2-aminofluorene. In the presence of 8 mM glycyrrhizic acid, the apparent Km and Vmax values were 0.15 +/- 0.01 mM and 0.95 +/- 0.11 nmol min(-1) mg(-1) protein, respectively, for 2-aminofluorene. In intact bacteria, the apparent Km and Vmax values were 1.28 +/- 0.48 mM and 4.08 +/- 1.06 nmol min(-1) 10(-10) colony forming units, respectively, for 2-aminofluorene. However, in the presence of 8 mM glycyrrhizic acid, the apparent Km and Vmax values were 0.67 +/- 0.09 mM and 1.82 +/- 0.37 nmol min(-1) 10(-10) colony forming units, respectively, for 2-aminofluorene. Taking these results together, the NAT activity in K. pneumoniae was inhibited by glycyrrhizic acid both in cytosol and intact bacteria. This study provides the first evidence to demonstrate that glycyrrhizic acid inhibits bacterial NAT activity. PMID- 9418947 TI - Development of bait formulations for control of intermediate hosts of African schistosome species. AB - Exploration of methods of snail control indicated a need for a new method because many failures in control programmes are due to lack of contact between molluscicides and the target snail population. The incorporation of molluscicides inside food pellets that are attractive to and ingestible by the target snails is suggested. Different concentrations of alcoholic extracts of Ambrosia maritima, Cucumis prophetarum and Rhynchosia minima-molluscicides of plant origin-were compared with niclosamide (Bayluscide) which is a strong synthetic molluscicide. They were incorporated into attractive food pellets and the lethal doses were determined. The results showed that Biomphalaria alexandrina snails are sensitive to only bait formulations with low concentrations of molluscicides. Laboratory and semi-field trials were conducted to study the efficacy of the prepared bait formulations on Biomphalaria alexandrina snails. The stability of these bait formulations was studied and it was observed that their effectiveness was reduced after long storage periods (> 3 weeks). The results revealed that Ambrosia maritima was the most effective molluscicide. PMID- 9418948 TI - In vitro and in vivo evaluations of the methaemoglobinaemic potential of xylidine isomers in the rat. AB - The objective of the present study was to evaluate the methaemoglobinaemic potential of the six isomers of xylidine (XYL) by two approaches: in vitro, using rat red blood cells and hepatic post-mitochondrial fractions in a two compartmental dialysis system; and in vivo, following a single oral dose of 4.8 mmol kg(-1) (p.o.) of each of the six XYL isomers. The in vitro experiments showed that all six XYL isomers at 1 mM concentration induced significant methaemoglobinaemia in the presence of active hepatic fractions, whereas non bioactivated XYL isomers were totally inactive. At lower incubation concentrations (0.3 mM and 0.06 mM), 3,5-XYL was still active, whereas the other isomers were less potent (0.3 mM) or totally ineffective (0.06 mM). The in vivo experiment revealed that all XYL isomers, except 3,5-XYL, did not induce significant methaemoglobinaemia after a single oral dose of 4.8 mmol kg(-1). The maximal percentage of methaemoglobin was 31.3 +/- 1.5 in the 3,5-XYL-treated rats, whereas it never exceeded 3% in all the other treatment groups, indicating that 4.8 mmol kg(-1) (p.o.) is in fact a no-observable-adverse-effect level for these XYL isomers. The quantitative differences between in vivo and in vitro results may have been due to additional bioactivation pathways (N-hydroxylation or ring hydroxylation) mediated by high Km enzymes operative at the high incubation concentrations used in vitro. The results of the present study suggest that 3,5-XYL is likely to be the only active isomer in the Sprague-Dawley rat at low exposure levels. PMID- 9418949 TI - Effect of lipoic acid administration on gentamicin-induced lipid peroxidation in rats. AB - The intraperitoneal administration of gentamicin (100 mg kg[-1] day[-1]) to rats is associated with an increased production of malondialdehyde (MDA), which is an end product of lipid peroxidation in the kidney. The level of glutathione (GSH) and the activity of three antioxidant systems--superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx)--were also decreased in the kidney. The liver, however, did not show any such alterations. Gentamicin (100 mg kg[-1] day[-1]) plus lipoic acid administration (25 mg kg[-1] day[-1]) by gastric intubation brought about a decrease in the degree of lipid peroxidation. An increase in the GSH level and in the activity of SOD, CAT and GPx was also observed. From these observations it can be concluded that administration of DL alpha-lipoic acid prevents lipid peroxidation, which may, at least partly, play an important role in the injury cascade of gentamicin-induced nephrotoxicity. PMID- 9418950 TI - Influence of atenolol and/or metformin on glutathione and magnesium levels in diabetic rats. AB - Recently there has been growing interest in studying the differences between different classes of antihypertensive drugs in preventing cardiovascular events in diabetic patients. Hypomagnesemia is common in diabetes mellitus, and correlates to its chronic complications and the associated alteration of the antioxidant enzyme activity. Depletion of reduced glutathione (GSH) in the blood has been demonstrated with myocardial injuries associating hypomagnesemia. A previous study has demonstrated a beneficial effect of metformin hydrochloride (Met), an antihyperglycemic drug, on both magnesium (Mg) and GSH levels in diabetic animals. The purpose of this study was to investigate the effect of oral atenolol, metformin (50 and 60 mg kg[-1] day[-1], respectively) and their combination for 14 days on Mg and GSH levels in blood, liver and heart of diabetic male Wistar rats, as these two parameters have been shown to be altered in diabetics and linked to myocardial ischemic injuries. The results of this investigation showed a state of low levels of Mg and GSH in both blood and liver of the diabetic animals. Treatment with atenolol alone did not change these levels significantly, however administration of metformin or atenolol/metformin increased significantly the GSH levels in both liver and blood, and returned the liver Mg content back to normal values. PMID- 9418951 TI - Effect of topically applied sulphur mustard on guinea pig liver. AB - The study revealed that topically applied sulphur mustard, which is a potent blistering agent with mutagenic and carcinogenic properties, is also hepatotoxic. It produces severe steatosis and other pathological alterations, accompanied by biochemical changes. There is a significant rise in the levels of glutamic oxaloacetic transaminase (GOT) and glutamic pyruvate transaminase (GPT) after exposure. The liver injury appeared to peak on the third day. Recovery at the ultrastructural level could be noticed on the sixth day but was far from complete because the gross pathological changes were still severe. PMID- 9418952 TI - Dose-dependent inhibition of phospholipase A2 by paraoxon in vitro: preliminary results. AB - To establish the dose dependency of phospholipase A2 (PLA2) inhibition by the organophosphorus compound (OPC) paraoxon (POX), human platelet membranes were incubated after Ca2+ removal (to inactivate the PLA2) with 0.3, 1 and 3 microg ml(-1) POX for 5, 30 and 60 min each. The PLA2 activity (pmol mg[-1] protein min[ 1]) was measured after subsequent enzyme reactivation. The PLA2 activity in native platelets was considered to be 100%; all other measured values are expressed as a percentage thereof. Data were analysed with the Mann-Whitney Wilcoxon rank order test and ANOVA. Statistical significance was assumed for P < or = 0.01. Paraoxon inhibited in a dose-dependent manner the PLA2 activity. Different incubation times of the inactive PLA2 with POX did not have any additional effect on the activity reduction after activation. At the tested POX concentrations the PLA2 activity was 42 +/- 5.4%, 29 +/- 3.4% and 15 +/- 6.6%, respectively. The corresponding butyrylcholine esterase (BChE) activities were <<1% of the baseline activity. Phospholipase A2 is less sensitive to POX inhibition than BChE and, at clinically achievable POX concentrations, shows a clear dose dependency. Further work is needed to elucidate the exact mechanism and time dependency of the phenomenon. PMID- 9418953 TI - Nickel and some nickel compounds. PMID- 9418954 TI - Redefining the lipophilin family of proteolipid proteins. AB - The past few years have seen a dramatic increase in our understanding, in molecular terms, of the involvement of the central nervous system proteolipid protein in myelinogenesis and X-linked genetic diseases. In addition, we have expanded our knowledge of the proteins that have been recruited into the vertebrate myelin membrane over the past 400 million years with the molecular cloning of several cDNAs encoding proteins which are homologous to the proteolipid protein gene. In searching for a name to distinguish these proteins from other "proteolipid" proteins of nonneural origin I propose that we resurrect the term "lipophilins" which describes a small family of unusually hydrophobic integral membrane proteins exhibiting identical topologies and similar physical properties. Two subgroups are distinguishable among the lipophilins based on the patterns of expression during development and the presence or absence of a small motif that is exposed to the extracellular space. PMID- 9418955 TI - Do olfactory glia have advantages over Schwann cells for CNS repair? AB - To a large extent the success of axon regeneration and sustained remyelination which distinguishes the PNS from the CNS is attributable to differences in their respective glial environments. For this reason, many have been attracted to the idea that repair of the CNS might be achieved by transplanting Schwann cells into areas of CNS pathology. Schwann cells will not only promote regeneration but will also myelinate axons thereby making them an appropriate cell type to mediate repair of lesions characterised by demyelination as well as axotomy. The recent discovery that olfactory glia are capable of forming myelin sheaths, together with their well-documented ability to support axon regeneration, means that these cells have a range of repair properties similar to that of Schwann cells. It is not clear at present which of these two alternatives, the Schwann cells or the olfactory glial cell, would be of greater benefit for achieving regeneration of axons or remyelination of persistent demyelination following transplantation into the CNS. In this article we review the repair properties of olfactory glia and identify the areas in which their use for repairing the CNS may have advantages over Schwann cells. PMID- 9418956 TI - Multiple isoforms of neuregulin are expressed in developing rat dorsal root ganglia. AB - Accumulating evidence suggests that neuregulin (NRG) plays special roles in the development of the mammalian nervous system. We have already identified NRG as a survival factor for Schwann cells during development. In this report, we have studied all possible NRG isoforms and expression of NRG in the developing rat dorsal root ganglia (DRG) and compared them with those of brain and spinal cord. Neural NRG isoforms comprise common immunoglobulin and epidermal growth factor domains. Various different transcripts were characterized, which arose by alternative splicing in several regions: N-terminal (exon 1 or 2), spacer (exon 5), juxtamembrane (exon 9 or 10), and cytoplasmic (exon 12, 13, or 14) domains. At least 13 novel isoforms among 16 splice variants were identified. The transmembrane isoforms of NRG are dominant forms in developing rat DRG. The mRNA expression of NRG isoforms in DRG is similar to that in spinal cord, while in brain the expression is much less. The mRNA in DRG was found at similar levels from birth to postnatal day 7 of the premyelinating stage, and it decreased afterward. Our results suggest that several NRGs, including isoforms not reported before, play a role as survival factors for Schwann cells in the premyelinating stage. PMID- 9418957 TI - Helix-loop-helix proteins in Schwann cells: a study of regulation and subcellular localization of Ids, REB, and E12/47 during embryonic and postnatal development. AB - Although basic helix-loop-helix (bHLH) proteins play an important role in transcriptional control in many cell types, the role of HLH proteins in Schwann cells has yet to be assessed. In this study, we have analyzed the expression of the dominant negative HLH genes, Id1 to Id4 and the class A gene REB, during Schwann cell development. We found that mRNA derived from these genes was present in the Schwann cell lineage throughout development including embryonic precursors and mature cells. The mRNA levels were not significantly regulated during development. Nevertheless, by using antibodies against the four different Id proteins, we found clear regulation of some of these genes at the protein level, in particular Id 2, 4, and REB, both in amount and nuclear/cytoplasmic localization. All these proteins are found in the nuclei of Schwann cell precursors but are not seen in nuclei of Schwann cells of newborn nerves. We observed extensive overlap in Id expression, especially in Schwann cell precursors that co-expressed all four Id proteins and REB. We also showed that Id 1 and 2 were up-regulated as Schwann cells progressed through the cell cycle. These data indicate that HLH transcription factors act as regulators of Schwann cell development and point to the existence of as yet unidentified cell type specific bHLH proteins in these cells. PMID- 9418959 TI - Oligodendrocyte-specific protein (OSP) is a major component of CNS myelin. AB - An oligodendrocyte-specific protein (OSP) cDNA was recently identified and found to be expressed primarily in oligodendrocytes and has a deduced amino acid sequence similar to that of peripheral myelin protein 22 (PMP-22). We raised antibodies against a synthetic peptide corresponding to OSP amino acid residues 179-194 which reacted with a 22 kd protein in mouse CNS. OSP immunoreactivity localized to spinal cord white matter tracts using immunohistochemistry in a similar distribution to that of MBP. OSP localized to CNS myelin biochemically with more than a 30-fold enrichment measured in purified myelin. We further purified the proteolipid fraction of myelin and determined that OSP contributes approximately 7% of total myelin protein making it the third most abundant protein in CNS myelin. No binding was found to several agglutinins or a HNK1 specific antibody suggesting that OSP is not a glycoprotein. PMID- 9418958 TI - Differential regulation of the zinc finger genes Krox-20 and Krox-24 (Egr-1) suggests antagonistic roles in Schwann cells. AB - Krox-20 and Krox-24 (Egr-1) encode closely related zinc finger transcription factors, which interact with the same DNA target sequences. Krox-20 is required for myelination in the peripheral nervous system. Using lacZ knock-in mutant mouse lines as well as immunohistochemical analyses, we have studied the expression of Krox-20 and Krox-24 in the Schwann cell lineage during normal development and following nerve lesion in the mouse and in human neuropathies. During embryogenesis, the two genes are expressed in a successive and mutually exclusive manner, Krox-24 being restricted to Schwann cell precursors and Krox-20 to mature Schwann cells. At birth, Krox-24 is reactivated and the two genes are coexpressed. In the adult, Krox-20 is expressed in myelinating cells, while Krox 24 is restricted to nonmyelinating cells. Following nerve lesion, Krox-24 is strongly induced in Schwann cells, reinforcing the link between its expression and the nonmyelinating and/or proliferative state, whereas Krox-20 is downregulated. These data are consistent with Krox-20 and Krox-24 playing antagonistic roles during the development of the Schwann cell lineage. In particular, their balance of expression might participate in the choice between myelinating and nonmyelinating pathways. PMID- 9418960 TI - Expression of insulin-like growth factor-binding protein messenger RNAs in developing rat oligodendrocytes and astrocytes. AB - Insulin-like growth factors, IGF-I and IGF-II, are potent regulators of oligodendrocyte development. Most of the IGF present in vivo is bound to members of a family of six high-affinity IGF-binding proteins (IGFBPs), which can either potentiate or inhibit IGF action, depending on other conditions. Additionally, serum contains a structurally unrelated protein, acid-labile sub-unit (ALS), which forms a ternary complex with IGF and IGFBP3. In this study, we used reverse transcriptase polymerase chain reaction (RT-PCR) to examine the expression of mRNAs for IGFBP 1-6 and ALS in purified populations of oligodendroglial cells and astrocytes. We found that astrocytes express all six IGFBPs. A2B5+/O4- oligodendrocyte precursors, O4+/O1- intermediate precursors, and O1+ oligodendrocytes express IGFBP3, 5, and 6, while IGFBP4 is expressed in oligodendrocyte precursors but not at more mature stages. We were unable to detect ALS mRNA in whole brain or in cultured oligodendroglial cells. The presence of differentially expressed IGFBPs in developing oligodendrocytes and astrocytes could significantly affect the biological activity of IGF-I and IGF-II in the central nervous system and the IGF-responsiveness of the IGFBP-expressing cells. PMID- 9418961 TI - Nerve growth factor signal transduction in mature pig oligodendrocytes. AB - It has previously been shown that nerve growth factor (NGF) is of functional significance for mature pig oligodendrocytes (OLs) in culture. The present data give evidence for the expression of TrkA, the so-called high-affinity NGF receptor, and of p75NTR, the so-called low-affinity NGF receptor. TrkA is upregulated during culturing, in contrast to the p75 receptor. Exposure of OLs to NGF induces an autophosphorylation of TrkA via its intrinsic tyrosine kinase. K 252a inhibits the TrkA autophosphorylation, which reduces the OL process formation to control levels. To the tyrosine-phosphorylated sites of TrkA several proteins, such as phospholipase C-gamma1, the adaptor protein SHC, the phosphotyrosine phosphatase SH-PTP2 (SYP) associate via their SH2 phosphotase SH PTP2 domain. The association of SHC to TrkA is shown by co-immunoprecipitation. Indirect evidence for a possible activation of PLC-gamma1 is given by an NGF induced increase of oligodendroglial [Ca2+]i. Downstream from TrkA, a mitogen activated protein kinase cascade, which includes Erk1 and Erk2, is operating. An in-gel myelin basic protein kinase assay revealed that NGF activates predominantly Erk1. Finally, it is shown that NGF stimulates expression of c-fos. PMID- 9418962 TI - Acetylcholine agonists stimulate mitogen-activated protein kinase in oligodendrocyte progenitors by muscarinic receptors. AB - Oligodendrocytes, the myelin-producing cells of the central nervous system, express muscarinic acetylcholine receptors (mAChR). Activation of this neurotransmitter receptor by the stable acetylcholine analog carbachol (CCh) triggers transducing events, modulating c-fos expression and cellular proliferation. To elucidate the signal transduction pathways involved in the transmission of these cellular events, we examined the ability of CCh to activate mitogen-activated protein kinase (MAPK) in primary cultures of oligodendrocyte progenitors prepared from newborn rat brain. CCh produced a concentration- and time-dependent increase in MAPK activity (predominantly the p42mapk or ERK2) as determined by in-gel MBP kinase assays. Using the non-selective muscarinic antagonist atropine we determined that MAPK-activation by CCH is mediated by muscarinic receptors. In the presence of PD098059, a specific inhibitor of MAPK kinase (MEK), MAPK activity was blocked. Similarly, the presence of extracellular calcium was required for CCh-mediated MAPK activation. To further elucidate the mechanisms involved in MAPK activation by CCh, the role of PKC was studied. In cells in which protein kinase had been downregulated by chronic treatment with 12 O-tetradecanoylphorbol 13-acetate (TPA), the effect of carbachol on MAPK activation was maintained. In contrast, the response to CCh was blocked by the PKC inhibitors H7 and bisindolylmaleimide GF109203X. Our results suggest that MAPK is implicated in the transmission of the signal for mACh receptors and involves a TPA-insensitive PKC pathway. Further work is required to define the upstream and downstream events which result in CCh-mediated MAPK activation and proliferation of oligodendrocyte progenitors. PMID- 9418964 TI - Expression of molecular chaperones and vesicle transport proteins in differentiating oligodendrocytes. AB - The major stages of oligodendrocyte differentiation are defined by the presence or absence of certain myelin-specific proteins. Events leading to the successful processing of these proteins, such as the folding, assembly, and trafficking of these proteins through the biosynthetic pathway, are largely undefined. In the present study, we have examined both cultured primary oligodendrocytes and immortalized oligodendrocyte cell lines for the presence of molecular chaperones and/or vesicle transport proteins. We find that a select set of these proteins are expressed relatively early in oligodendrocyte differentiation, whereas a characteristically different set of proteins are expressed at later stages of oligodendrocyte differentiation. In other systems, these proteins participate in the folding and assembly of protein complexes, in the prevention of protein aggregation, as well as the trafficking of proteins via vesicles to specific subcellular destinations including the plasma membrane. Some of the chaperones and/or vesicle transport proteins investigated in this study may play a pivotal role in the certain aspect of myelin biogenesis. PMID- 9418963 TI - Effect of neu differentiation factor isoforms on neonatal oligodendrocyte function. AB - Previous studies have suggested that neu differentiation factor (NDF), a member of the neuregulin (NRG) family of growth factors, may regulate the development of PNS and CNS glial cells. There is limited information concerning the potential role of NDF on the development of neonatal (immature) oligodendrocytes (OLG) into adult OLG. We now report the effect of the two major isoform families of NDF (NDF alpha and NDF beta) on the development of cultured rat neonatal OLG. Immunocytochemical and western blot analyses of neonatal OLG using anti-erb-B antibodies revealed that these immature OLG express all four members of NRG (erb B) receptors. Treatment of neonatal OLG with varying concentrations of either NDF alpha or NDF beta did not have a mitogenic effect on cultured neonatal OLG. Pretreatment of immature OLG with either of the NDF isoforms also did not influence the subsequent mitogenicity of other known OLG mitogens. However, treatment of neonatal OLG with either isoform of NDF influenced the survival of these cells by protecting the cells from apoptosis. Additionally, treatment of neonatal OLG with either NDF alpha or NDF beta resulted in more extensive process formation compared to control, non-treated OLG. PMID- 9418965 TI - Partial characterization of the 5'-flanking region of trout IP: a Po-like gene containing a PLP-like promoter. AB - The IP gene of trout encodes two Po-like glycoproteins which are expressed by oligodendrocytes in the fish CNS. A 679 bp fragment of its 5'-flanking region was isolated from a genomic library and sequenced. The transcription start point was determined 124 bp upstream the ATG initiator codon by primer extension analysis. Apart from a modified TATA-box and an inverted CCAAT-box located at canonical distances from the transcription start site several eucaryotic cis-acting regulatory elements were identified in the 679 bp upstream region, including an AP-1 binding site, a brain specific Sp1 motif, a cyclic AMP responsive element and a consensus sequence for POU homeodomain protein binding. The occurence of respective DNA-binding proteins for Sp1, AP-1 and POU in the nuclei of trout oligodendrocyte progenitor cells was verified by gel retardation experiments. Functional activity of various subfragments of the 679 bp upstream region was demonstrated by CAT reporter gene analysis. A computer-assisted sequence alignment of the trout IP 5'-flanking end with the corresponding region of the mammalian PLP gene promoter revealed four sites of high homology, while similarity with the mammalian Po gene promotor was low. The results are discussed with respect to the phylogenetic shift from Po-like proteins to PLP during evolution of the vertebrate CNS myelin sheath. PMID- 9418966 TI - Differentiating oligodendrocytes inhibit neuronal growth cone motility in different ways. AB - Oligodendrocyte-specific proteins are thought to limit regeneration by inhibiting motility of the neuronal growth cone. These inhibitory proteins are among an array of molecules, each expressed in a stage-specific fashion as an oligodendrocyte differentiates. Here we show that neuronal growth cone response to contact with differentiating oligodendrocytes does not strictly correlate with the expression of inhibitory proteins. This suggests that the neuronal growth cone response depends on the context in which inhibitory molecules are encountered. In addition, we provide evidence that there are at least two cellular pathways, within the same growth cones, that lead to growth cone collapse. PMID- 9418967 TI - Apoptosis of microglia and oligodendrocytes after spinal cord contusion in rats. AB - Following spinal cord contusion in the rat, apoptosis has been observed in the white matter for long distances remote from the center of the lesion and is primarily associated with degenerating fiber tracts. We have previously reported that many of the apoptotic cells are oligodendrocytes. Here we show that the oligodendrocyte death is maximal at 8 days postinjury and suggest that loss of oligodendrocytes may result in demyelination of axons that have survived the initial trauma. There are two mechanisms that may account for the observed oligodendrocyte apoptosis. The apoptotic cell death may result from the loss of trophic support after axonal degeneration or it may be the consequence of microglial activation. The hypothesis that oligodendrocyte apoptosis is secondary to microglial activation is supported by our observations of microglia with an activated morphology in the same regions as apoptosis and apparent contact between some of the apoptotic oligodendrocytes and microglial processes. In addition to oligodendrocyte apoptosis, a subpopulation of microglia appears to be susceptible to apoptotic cell death as well, as evidenced by the presence of apoptotic bodies in OX42 immunopositive profiles. Thus, the population of apoptotic cells following spinal cord contusion is comprised of oligodendrocytes and putative phagocytic microglia or macrophages. Given the delayed time course of oligodendrocyte death, the apoptotic death of oligodendrocytes may be amenable to pharmacological intervention with subsequent improvement in functional recovery. PMID- 9418968 TI - Delayed and incomplete myelination in a transgenic mouse mutant with abnormal oligodendrocytes. AB - In search of animal models suitable for investigating myelin repair, we have analysed myelinogenesis in a transgenic mouse mutant with delayed myelination, but with a normal life-span. The 2-50 mutant which carries a c-myc gene under the regulation of the myelin basic protein promoter has been described previously (Orian et al.: J Neurosci Res 39:604-612, 1994). Here we show that appropriate mRNA transcripts and their corresponding protein products are generated, but that the accumulation of these products is delayed in transgenic mice with respect to nontransgenic littermates. This phenomenon is associated with aberrant myelin and paucity of normal oligodendrocytes. Myelination appears to be carried out by abnormal, oligodendrocyte-like cells. We propose that the primary defect in the 2 50 mutant is an inability to generate the normal number of mature oligodendrocytes. This mutant represents a novel class of mutant in which oligodendrocyte development and myelination can be studied in the absence of interference with a gene for a structural protein of myelin, in an animal with normal survival. It may also represent a new tool to investigate in vivo gliogenesis and regulatory events bringing about the coordinated regulation of myelin protein synthesis. PMID- 9418969 TI - Abnormal nerve conduction studies in mice expressing a mutant form of the POU transcription factor SCIP. AB - We have previously described transgenic mice that harbor a dominant-negative antagonist of the POU protein SCIP (termed deltaSCIP). Native SCIP is expressed in promyelinating Schwann cells, where it represses expression of the myelin structural genes. The deltaSCIP mice display morphologic and behavioral abnormalities, including decreased axonal diameter, increased myelin thickness, developmentally early myelination, and clinical features of neuropathy. To assess the neurophysiologic correlates of these abnormalities, a series of electrophysiologic tests was performed. Despite having smaller diameter axons, mice expressing the deltaSCIP transgene had similar maximum conduction velocities in caudal, sural, and tibial nerves compared to wild-type controls. Therefore, conduction in deltaSCIP animals was faster than predicted by axon diameter alone. Compound amplitude responses were 38% higher in the deltaSCIP caudal nerve. DeltaSCIP tibial F-wave responses showed less difference between minimum and maximum latencies than controls, suggesting less variance between fastest and slowest conducting fibers. These data further characterize the functional components of the deltaSCIP phenotype. In addition, these studies address the physiologic sequelae of altering the g-ratio in the absence of demyelination or axonal degeneration. PMID- 9418970 TI - Targeted inactivation of the X-linked adrenoleukodystrophy gene in mice. AB - In its severe form, X-linked adrenoleukodystrophy (ALD) is a lethal neurologic disease of children, characterized by progressive cerebral demyelination and adrenal insufficiency. Associated with a biochemical defect of peroxisomal beta oxidation, very long-chain fatty acids (VLCFA) build up in tissues that have a high turnover of lipids, such as central nervous system (CNS) white matter, adrenal cortex, and testis. Whether the abnormal accumulation of VLCFA is the underlying cause of demyelination or merely an associated biochemical marker is unknown. ALD is caused by mutations in the gene for a peroxisomal membrane protein (ALDP) that shares structural features with ATP-binding-cassette (ABC) transporters. To analyze the cellular function of ALDP and to obtain an animal model of this debilitating disease, we have generated transgenic mice with a targeted inactivation of the ald gene. Motor functions in ALDP-deficient mice developed at schedule, and unexpectedly, adult animals appeared unaffected by neurologic symptoms up to at least 6 months of age. Biochemical analyses demonstrated impaired beta-oxidation in mutant fibroblasts and abnormal accumulation of VLCFAs in the CNS and kidney. In 6-month-old mutants, adrenal cortex cells displayed a ballooned morphology and needle-like lipid inclusions, also found in testis and ovaries. However, lipid inclusions and demyelinating lesions in the CNS were not a feature. Thus, complete absence of ALDP expression results in a VLCFA storage disease but does not impair CNS function of young adult mice by pathologic and clinical criteria. This suggests that additional genetic or environmental conditions must be fulfilled to model the early-onset and lethality of cerebral ALD in transgenic mice. PMID- 9418971 TI - Intracellular transport of the DM-20 bearing shaking pup (shp) mutation and its possible phenotypic consequences. AB - Paralytic tremor (pt) in rabbits and shaking pup (shp) in dogs are allelic dysmyelinated mutants of the proteolipid protein (Plp) gene. Both mutations affect the same amino acid, histidine36, which is replaced by glutamine in pt and by proline in shp. Phenotypic expression of these two mutations is very different. Paralytic tremor presents a much milder form of dysmyelination than shaking pup. The number of oligodendrocytes in the mutant rabbit is normal, while in the dog, the oligodendrocyte number is reduced due to early death or incomplete maturation. We have previously reported an abnormal intracellular transport of the PLPpt, whereas DM-20pt was normally transported to the cell membrane. In the present study, we show that the transport of the two isoforms containing the shp mutation is impaired in transfected Cos-7 cells. Cotransfecting cells with different ratios and combinations of mutated PLP and DM 20 cDNAs, we demonstrated that DM-20pt, but not DM-20shp, facilitates intracellular trafficking and integration into the plasma membrane of either of the two mutated PLPs. The phenotypic difference between these two allelic mutations can result from differences in DM-20 protein trafficking and sorting. These results show that the loss of function of PLP is not position-dependent but depends on the nature of the mutation. PMID- 9418973 TI - Isolation and transplantation of multipotential populations of epidermal growth factor-responsive, neural progenitor cells from the canine brain. AB - Glial cell transplantation into myelin-deficient rodent models has resulted in myelination of axons and restoration of conduction velocity. The shaking (sh) pup canine myelin mutant is a useful model in which to test the ability to repair human myelin diseases, but as in humans, the canine donor supply for allografting is limited. A solution may be provided by self-renewing epidermal growth factor (EGF)-responsive multipotential neural progenitor cell populations ("neurospheres"). Nonadherent spherical clusters, similar in appearance to murine neurospheres, have been obtained from the brain of perinatal wildtype (wt) canine brain and expanded in vitro in the presence of EGF for at least 6 months. Most of the cells in these clusters express a nestin-related protein. Within 1-2 weeks after removal of EGF, cells from the clusters generate neurons, astrocytes, and both oligodendroglial progenitors and oligodendrocytes. Transplantation of lacZ expressing wt neurospheres into the myelin-deficient (md) rat showed that a proportion of the cells differentiated into oligodendrocytes and produced myelin. In addition, cells from the neurosphere populations survived at least 6 weeks after grafting into a 14-day postnatal sh pup recipient and at least 2 weeks after grafting into an adult sh pup recipient. Thus, neurospheres provide a new source of allogeneic donor cells for transplantation studies in this mutant. PMID- 9418972 TI - Inflammation promotes survival and migration of the CG4 oligodendrocyte progenitors transplanted in the spinal cord of both inflammatory and demyelinated EAE rats. AB - Oligodendrocyte progenitor CG4 cells were labeled with bisbenzimide and transplanted in the lumbar spinal cord of rats 15 to 17 days prior to the induction of experimental autoimmune encephalomyelitis (EAE). EAE was induced by immunization with the encephalitogenic peptide of myelin basic protein (amino acids 68-88; C1) in adjuvant, either alone or in combination with a single injection of an anti-myelin oligodendrocyte glycoprotein (MOG) antibody to enhance central nervous system (CNS) demyelination. In control animals without EAE, the survival and migration capacity of CG4 cells was minimal. In striking contrast, both the survival and migration of this oligodendrocyte progenitor cell line were greatly enhanced in animals with EAE. In both disease models, large number of CG4 cells were still found in the spinal cord 50 days after transplantation, by which time they had migrated up to 6 cm from the transplantation site. Migrating CG4 cells were found in the subpial space, around the ependyma and blood vessels, and as well as in the grey and white matter of the CNS parenchyma. In all these locations, the CG4 cells were often associated with reactive astrocytes. These data strongly support the concept that inflammatory responses within the CNS promote, rather than inhibit, the survival and migration of transplanted oligodendrocyte progenitors in the adult CNS. PMID- 9418974 TI - Transplantation of CG4 oligodendrocyte progenitor cells in the myelin-deficient rat brain results in myelination of axons and enhanced oligodendroglial markers. AB - Transplantation of oligodendrocyte (Ol) progenitor cells into the central nervous system is a promising approach for the treatment of myelin disorders. This approach requires providing adequate numbers of healthy cells with myelinating potential. We recently showed the successful transplantation of Ol progenitors into the myelin-deficient (md) rat brain. In the present work, CG4 cells, a cell line with properties of Ol progenitors, were labeled with fast blue and grafted into P3-P5 pups born to carrier mothers. Examination of host brains 2 weeks posttransplant indicated that CG4 cells display a much more extensive migration capacity than their wild-type counterparts. These cells synthesized myelin components. In addition, ultrastructural analysis showed myelin formation along axons of md hosts in various brain regions, including corpus callosum, cerebellum, and brainstem. Furthermore, in situ hybridization studies performed on sagittal sections revealed extensive expression of transferrin-mRNA within the md host parenchyma. The high survival and functional features displayed by CG4 cells after transplantation, together with their striking wide distribution within the host parenchyma, as assessed by the presence of myelinated fibers in mutant hosts, emphasizes the importance of using highly motile and proliferative Ol progenitor cells. Strategies to improve the condition and life span of md rat pups are currently under investigation. PMID- 9418976 TI - Capacitation status and fertility of fresh and frozen-thawed ram spermatozoa. AB - The effect of cryopreservation on the capacitation status and fertility of ram spermatozoa was observed. After the chlortetracycline staining technique was validated for ram spermatozoa, it was applied to fresh or long-term frozen-stored spermatozoa. Fresh spermatozoa displayed mainly the F pattern (non-capacitated; 61.3%), becoming B pattern (capacitated; 54%) and AR pattern (acrosome reacted; 41%) with incubation (6 h at 37 degrees C). In contrast, frozen spermatozoa displayed the B pattern (65.9%), becoming the AR pattern (64.2%) with incubation. This demonstrates that cryopreservation may cause membrane changes in ram spermatozoa functionally equivalent to capacitation. The differences in capacitation status did not affect in vitro fertilization rates between fresh and frozen spermatozoa, but pregnancy rates at Day 18 after intrauterine artificial insemination were higher for fresh than for frozen spermatozoa. This difference was not evident at Day 50, possibly as a result of the high embryonic loss between Days 18 and 50 when fresh unincubated and frozen incubated spermatozoa were inseminated. Further research is necessary to determine what part of the cryopreservation process is responsible for the membrane changes in ram spermatozoa. PMID- 9418975 TI - Influence of IN-1 antibody and acidic FGF-fibrin glue on the response of injured corticospinal tract axons to human Schwann cell grafts. AB - Two strategies have been shown by others to improve CST regeneration following thoracic spinal cord injury: 1) the administration of a monoclonal antibody, IN 1, raised against a myelin-associated, neurite growth inhibitory protein, and 2) the delivery of acidic fibroblast growth factor (aFGF) in fibrin glue in association with peripheral nerve grafts. Because autologous transplantation of human Schwann cells (SCs) is a potential strategy for CNS repair, we evaluated the ability of these two molecular agents to induce CST regeneration into human SC grafts placed to span a midthoracic spinal cord transection in the adult nude rat, a xenograft tolerant strain. IN-1 or control (HRP) antibodies were delivered to the injury/graft region by encapsulated hybridoma cells ("IN-1 ravioli") or daily infusion of hybridoma culture supernatant; aFGF-fibrin glue was placed in the same region in other animals. Anterograde tracing from the motor cortex using the dextran amine tracers, Fluororuby (FR) and biotinylated dextran amine (BDA), was performed. Thirty-five days after grafting, the CST response was evaluated qualitatively by looking for regenerated CST fibers in or beyond grafts and quantitatively by constructing camera lucida composites to determine the sprouting index (SI), the position of the maximum termination density (MTD) rostral to the GFAP-defined host/graft interface, and the longitudinal spread (LS) of bulbous end terminals. The latter two measures provided information about axonal die-back. In control animals (graft only), the CST did not enter the SC graft and underwent axonal die-back [SI = 1.4 +/- 0.1, MTD = 2.0 +/- 0.2, LS = 1.3 +/- 0.3, (n = 3)]. Results of IN-1 delivery from ravioli did not differ from controls, but injections of IN-1-containing supernatant resulted in a significant degree of sprouting but did not prevent axonal die-back [SI = 1.9 +/- 0.1, MTD = 1.5 +/- 0.2, LS = 1.1 +/- 0.1, (n = 7)] and traced fibers did not enter grafts. Acidic FGF dramatically reduced axonal die-back and caused sprouting [SI = 2.0 +/ 0.1 (n = 5), MTD = 0.5 +/- 0.04 (n = 6), LS = 0.4 +/- 0.1 (n = 6)]. Some traced fibers entered SC grafts and in 2/6 cases entered the distal interface. We conclude that 1) human SC grafts alone do not support the regeneration of injured CST fibers and do not prevent die-back, 2) grafts plus IN-1 antibody-containing supernatant support some sprouting but die-back continues, and 3) grafts plus aFGF-fibrin glue support regeneration of some fibers into the grafts and reduce die-back. PMID- 9418977 TI - Ontogeny of thyroid hormone receptors in the brushtail possum (Trichosurus vulpecula). AB - Newborn marsupials do not have a thyroid gland at birth. The gland develops while the young marsupial is in the mother's pouch. The young brushtail possum initiates secretion of thyroid hormones from its own thyroid at about Day 65 post partum. However, during the first three weeks of pouch life thyroxine is passed from the mother to the young via the milk. To determine if this maternal thyroxine can effect organ development in the young possum before it initiates secretion of thyroxine from its own thyroid, the ontogeny of thyroid hormone receptors was determined in nuclear extracts of lung, liver and kidney by radioreceptor assay, using (125)I-labelled tri-iodothyronine as ligand. Receptor density was calculated for tissues removed from young possums at Days 25 (n = 5), 50 (n = 4), 100 (n = 3) and 150 (n = 4) and from adults (n = 5). Receptors were found in possums of all age groups, including the small 25-day pouch young. Significant differences were not found in the receptor density between different tissues or at various ages. The association constant Ka (4.0+/-2.6 L nmol[-1] for lung) was similar in different tissues and at the various ages examined. The passage of thyroid hormones from the mother to the developing marsupial via the milk may have a role in the slow development of organ systems early in pouch life by acting on thyroid receptors in the pouch young. However, the functional maturation of the thyroid gland of the young possum, not an increase in receptors, appears to coincide with the rapid increase in the rate of growth and development which occurs in later pouch life. PMID- 9418978 TI - In vivo effects of epidermal growth factor on epidermal pattern formation and hair follicle initiation in the marsupial bandicoot Isoodon macrourus. AB - The extrauterine development of marsupial pouch young (northern brown bandicoot Isoodon macrourus) has facilitated the study of the effects of murine epidermal growth factor (mEGF) on pattern formation in skin. Hair follicle initiation and development, which in the mouse would occur from about Days 13-14 of gestation onward, occurs postnatally. In the present study the effect in vivo of mEGF on developing skin corresponding to mouse gestational ages from Day 13 onward was examined. Subcutaneous injections of mEGF (0.5, 1.0 and 2.0 microg g[-1] body weight) or equivalent volumes of saline (0.9% w/w) were administered daily, before and during hair follicle initiation and development. Murine EGF inhibited the formation of hair follicles, hair follicle sweat glands, sebaceous glands and dermal papillae. The pattern of follicle initiation was perturbed. The characteristic trio follicle grouping was absent, and follicle rudiment densities (no. per mm2 skin surface) were significantly lower in animals treated with mEGF, whereas follicle diameters were increased. These data may reflect a role for the epidermal growth factor (EGF) receptor in epidermal pattern formation. The EGF receptor and its potential ligands (such as EGF, transforming growth factor (TGF alpha) or other yet-to-be-discovered ligands) perhaps act as parts of a pattern forming system in vertebrate skin. PMID- 9418979 TI - Effect of desialylation of highly purified isoforms of human luteinizing hormone on their bioactivity in vitro, radioreceptor activity and immunoactivity. AB - To establish whether sialic acid content is responsible for an observed 7-8-fold variability in bioactivity in vitro of highly purified human pituitary luteinizing hormone (hLH) isoforms, the bioactivity in vitro, radioreceptor activity and immunoactivity of hLH isoforms were determined before and after enzymatic desialylation. Three immunofluorometric assays with different hLH specificities allowed characterization of 13-24 pituitary hLH isoform preparations of pI 7.03-8.98 in terms of sialic acid content (1-5 sialic acid residues per LH molecule), bioactivity in vitro (4030-30,000 I.U. mg[-1]), radioreceptor activity (6420-25,400 I.U. mg[-1]) and hLH immunoactivity (2900 4400 to 18,300-27,300 I.U. mg[-1]). Significant positive correlations between sialic acid content and either immunoactivity or in vitro bioactivity were observed, whereas radioreceptor activity showed a curvilinear response. Following more than 90% removal of sialic acid, both in vitro bioactivity and radioreceptor activity were increased, although specific activity still differed between isoforms; immunoactivities were unaffected. It is concluded that the presence of the sialic acid residue(s) on hLH isoforms does partially contribute to the in vitro bioactivity and radioreceptor activity of the isoforms, but that hLH immunoactivity is independent of sialic acid content. PMID- 9418980 TI - Influence of maternal bodyweight on size, conformation and survival of newborn lambs. AB - Although body condition score was not significantly different between light (<55 kg, n = 6) and heavy (> or =60 kg, n = 7) ewes at mating, it declined between Day 30 and Day 90 of gestation in light but not heavy ewes, and remained lower up to term. All ewes bore twins, delivered near term (Days 144-146) by Caesarean section. One lamb was immediately placed into a warm (30 degrees C; WD) and its twin into a cool (15 degrees C; CD) ambient temperature, and tissues were sampled at 0.5 h or 6 h. All CD lambs born to light ewes exhibited hypothermia and/or respiratory failure and did not survive longer than 30 min; these symptoms were not observed in their WD twins or any lamb born to heavy ewes. Total lamb birth weight, placental weight and fetal cotyledonary weight were lower with light than with heavy ewes. Lambs born to light ewes had less perirenal adipose tissue and smaller liver, heart, kidneys, brain, adrenals and thyroid, although their heart, brain and pancreas represented a larger proportion of total bodyweight; pancreas weight was similar to that in lambs born to heavy ewes. Hence, maternal bodyweight critically influences placental weight and lamb size and survival after birth. PMID- 9418981 TI - Role of glucose, fatty acids and protein in regulation of testicular growth and secretion of gonadotrophin, prolactin, somatotrophin and insulin in the mature ram. AB - This study tested whether the effects of nutrition on gonadotrophin secretion and testicular growth in mature rams are due to increases in the supply of glucose, fatty acids (FA) or amino acids. Responses to protein (casein) and glucose, alone or in combination, were compared with responses to lupin grain and responses to a combination of protein, glucose and FA (acetate, propionate and vegetable oil). Glucose and casein were infused intra-abomasally whereas lupins and FA were added to the diet. Lupin feeding decreased blood growth hormone (GH) concentrations, but increased pulsatile luteinizing hormone (LH) secretion and increased the concentrations of follicle-stimulating hormone (FSH), prolactin, glucose and insulin. These effects were associated with testicular growth. Glucose or casein increased insulin concentrations and decreased GH concentrations, but did not affect gonadotrophins or testicular growth. There was no synergism between casein and glucose. Responses elicited by adding FA to the glucose+casein treatment were similar to those observed with lupins. In conclusion, the reproductive axis does not seem to be closely linked with dietary intakes of amino acids or with circulating concentrations of glucose, insulin or GH. However, the energetic components of the diet, particularly the fatty acids, appear to play a key role in the reproductive responses to changes in nutrition. PMID- 9418982 TI - Regulation of bovine cervical secretion of prostaglandins and synthesis of cyclooxygenase by oxytocin. AB - Prostaglandin E2 (PGE2) can cause softening of the bovine cervix at oestrus when receptors for oxytocin (OT) are maximally present, indicating a relationship between OT and PGE2 production. It was therefore determined whether OT can stimulate prostaglandin synthesis or induce cyclooxygenase expression in cervical external os segments obtained from pre-oestrous-oestrous cows. Tissues were minced and incubated (50-100 mg mL[-1] 6 h[-1]) in the presence of OT (10 ng mL[ 1]), progesterone (P4) (5 ng mL[-1]) and/or indomethacin (5 microg mL[-1]). It was found that OT stimulated basal PGE2 (7.79+/-1.22 ng 100 mg[-1], mean+/ s.e.m.; n = 6) in external os segments from pre-oestrous-oestrous cows (P < 0.03), whereas P4 and indomethacin inhibited basal and OT-stimulated PGE2 production (P < 0.05). Basal prostaglandin F2alpha (PGF2alpha) production was minimal (<1 ng 100 mg[-1]) and OT had no effect on its production. Expression of cyclooxygenase was measured by Western blot analysis following incubation of the tissue (100 mg 1.5 mL[-1] 3 h[-1]) in the presence of OT (10 ng mL[-1]) and in the presence of P4 (5 ng mL[-1]). It was found that OT stimulated the induction of cyclooxygenase II (79+/-10%; n = 7, P < 0.05). In contrast, P4 inhibited the basal expression of this enzyme (-40+/-5%, n = 7, P < 0.05) in the presence or absence of OT. It is concluded that, in vitro, OT stimulates PGE2 synthesis by the bovine cervix at oestrus and that this effect is mediated by cyclooxygenase. PMID- 9418983 TI - Regulation of reproductive tract immunoglobulins by oestradiol-17beta in the European Red Fox. AB - The effect of the ovarian hormone, oestradiol-17beta, on reproductive tract immunity in the female fox was investigated. Reproductive tract antibody responses were induced by either Peyer's patch immunization with a recombinant fox sperm protein, or by oral immunization with live, attenuated Salmonella typhimurium. The effect of exogenous oestradiol-17beta or the stage of the oestrous cycle on reproductive tract immunity was assessed. The secretion of specific vaginal IgA, but not vaginal IgG, antibodies was reduced by exogenous treatment with oestradiol-17beta, while both specific vaginal IgA and vaginal IgG levels declined during the period of natural oestrus. It is concluded that oestradiol-17beta, and probably other reproductive hormones, are involved in the regulation of antibody-secretion in the fox reproductive tract, and that reproductive status is an important factor to consider in the design and application of vaccines which aim to induce immunity within the female reproductive tract. PMID- 9418984 TI - Parthenogenetic activation of pig eggs by exposure to protein kinase inhibitors. AB - The objective of the present study was to assess the effect of low concentrations of protein kinase inhibitors on activation. Pig eggs were electrostimulated or cultured with the following: 10 microM 1-[5-isoquinolinylsulfonyl]-2 methylpiperazine, HCl H7 for 24 h; 100 microM H7 for 24 h; 10 nM staurosporine for 24 h; or with 20 microM staurosporine for 20 min followed by Whitten's medium for 24 h. Rates of pronuclear formation in eggs (n = 1240) subjected to these treatments were: untreated, 6.2%; electrostimulated, 77.1%; 10 microM H7, 10.0%; 100 microM H7, 65%; 10 nM staurosporine, 24.2%; and 20 microM staurosporine, 67.3% (significance at P < or = 0.05: 10 microM H7 vs untreated, not significant; 20 microM staurosporine vs 100 microM H7, not significant). Percentages of eggs (n = 125) expressing a 22-kDa band after treatment were: untreated, 37.5%; electrostimulated, 100%; 10 microM H7, 72%; 100 microM H7, 66.7%; 10 nM staurosporine, 40.0%; and 20 microM staurosporine, 77.3% (significance at P < or = 0.10: 100 microM H7, 10 nM staurosporine and 20 microM staurosporine vs 10 microM H7, not significant; 100 microM H7 and 10 nM staurosporine vs untreated, not significant). Transmission electron microscopy of ultrathin sections of treated eggs revealed that cortical granules were present in over half the untreated eggs, as well as over half of the eggs treated with 100 microM H7 or 10 nM staurosporine; in contrast, all cortical granules were absent from electrically-activated eggs. The results indicate that long-term exposure of eggs to low concentrations of broad-spectrum protein kinase inhibitors induces some of the events commonly associated with fertilization. PMID- 9418985 TI - Angiotensin II as a semen extender component increases retention of spermatozoa within the uterus of the heifer. AB - The effects of angiotensin II (Ang II) as a semen extender were studied. In the first experiment, individual ejaculates from 10 bulls were split and extended in egg yolk citrate in the absence or presence of varying concentrations of Ang II (10[-5]-10[-10] M) to a final concentration of 35 x 10(6) sperm per mL. The percentage of intact acrosomes and percentage motility were determined in all treatments for all bulls at 0 h (immediately post thaw) and after incubation for 4 h at 37 degrees C. Extension of the semen with Ang II did not affect spermatozoal viability at either time studied. In the second experiment, mixed breed virgin heifers were induced into oestrus with intramuscular injections of prostaglandin F2alpha on Days 0 and 3. Animals that stood to be mounted were paired for bilateral intracornual insemination using a 0.5-mL French straw on each side approximately 8 h later. One of the paired heifers received semen containing Ang II (10[-5] M) while the other received control semen. A 1-mL aspirate of vaginal mucus was collected at hourly intervals for 8 h after insemination. Concentration of spermatozoa was determined by haemocytometry. There was a significant reduction in cumulative semen loss into the vagina of heifers inseminated with Ang II extended semen (14.4%) compared with heifers inseminated with control semen (19.7%). This suggests that Ang II, when added to extended semen, may reduce retrograde sperm loss following insemination without affecting sperm viability. PMID- 9418986 TI - Physiological limits to further improvement in the efficiency of oestrous synchronization in goats. AB - The variability between animals in the timing of oestrus after administration of a synchronization treatment seems to explain the low rate of fertility in goats inseminated at a predetermined time after progesterone withdrawal. Two experiments were performed during the breeding season to test whether the variation was due to the exogenous hormone regime or to the endogenous physiology of the animals. Twenty-one goats were given a synchronization treatment consisting of a vaginal sponge impregnated with 45 mg of fluorogestone acetate (FGA) for 11 days associated with intramuscular injection of 400 I.U. of equine chorionic gonadotrophin (eCG) and 50 microg of cloprostenol 48 h before sponge removal. Progesterone concentrations were measured during the subsequent cycle and the patterns were modelled to allow precise determination of the onset of luteolysis. Oestrus and the luteinizing hormone (LH) surge began 33.0+/-6.8 h and 76.0+/-33.0 h after sponge withdrawal, v. 43.4+/-5.7 h and 90.0+/-36.0 h after natural luteolysis. For both observations, the between-goat variability was larger during the natural than during the synchronized oestrus (P < 0.05). The duration of the oestrous cycle was independent of the number of corpora lutea (CL), whereas the duration of luteal phase was shorter in goats with 2-3 CL (16.4+/-0.9 day than in those with 1 CL: 17.7+/-1.3 day; P < 0.05). In the second experiment, 20 goats were ovariectomized and given a vaginal sponge as described above. Sixteen h after sponge removal, they were injected with 50 microg of oestradiol benzoate (ODB). This treatment was repeated with the second sponge being inserted 1-2 days after observation of oestrus. Oestrus and LH surge were observed: 32.8+/-6 8 h v. 27.8+/-7.8 h after the first ODB injection, and 36.6+/ 7.3 h v. 34.3+/-4.8 h after the second ODB injection. No relationship was observed between data of the two experiments. In both cases, the variability in the occurrence of oestrus and LH surge was of the same order as observed in the first experiment. This study shows that the timing of oestrus and LH surge is less variable after progestagen treatment than during a natural oestrous cycle. Moreover, a significant proportion of variability is inherent in the delays following the oestradiol peak, suggesting that further improvement in the synchronizing capacity of treatment based on progestagen administration is unlikely. PMID- 9418987 TI - Nitric oxide synthase regulation during embryonic implantation. AB - It has previously been demonstrated that uterine nitric oxide synthase (NOS) activity increases before embryonic implantation in rats. The aim of the present work was to investigate the regulation and the physiological relevance of the nitric oxide (NO) system in ovoimplantation. The increase in NOS activity in early pregnancy was found to be independent of the presence of embryos in the uterus. Whereas the Ca2+-dependent isoform of NOS increased gradually in the preimplantation days, the Ca2+-independent isoform increased just at the beginning of implantation (Day 5, 1800 hours); then the activity of both isoforms declined. Oestradiol, whose concentration peaks before implantation, might be regulating NOS activity in the uterus, since treatment of rats with tamoxifen, a receptor antagonist, reduces the activity of both isoforms to preimplantation levels. Intraluminal injections of L-NAME (0.5 mg kg[-1]), a competitive inhibitor of NOS, reduced by 50% the number of implanted embryos; this suggests that the NO system plays a role during implantation. The data suggest that oestradiol might be a modulator of NOS activity during nidation and that NO production is necessary to achieve a successful embryo implantation. PMID- 9418988 TI - Excretion profiles of some reproductive steroids in the faeces of captive Nepalese red panda (Ailurus fulgens fulgens). AB - Faecal samples were collected up to once daily from three female Nepalese red panda (Ailurus fulgens fulgens) prior to, during, and after the expected 1995 breeding season. Radioimmunoassay of faecal progestins and oestrogens showed hormone profiles that suggest that this species is a seasonally polyoestrous, induced ovulator. Examination of faecal extracts following high pressure liquid chromatography and using antisera of high specificity, demonstrated at least five faecal progestins but only one major oestrogen, probably oestradiol. Samples from males were collected up to once weekly during the breeding season in 1996, and androgens measured by radioimmunoassay. Faecal androgens fluctuated widely for all males. PMID- 9418990 TI - Perivascular spaces in the basal ganglia of the human brain: their relationship to lacunes. AB - There is evidence for lymphatic drainage of interstitial fluid from the brain along perivascular spaces in a number of mammalian species. Ultrastructural studies suggest that there are similar drainage pathways in the human cerebral cortex. Perivascular spaces in the basal ganglia, however, differ from those in the cortex in that they dilate to form lacunes and rarely accumulate beta-amyloid (amyloid angiopathy) in Alzheimer's disease; in the cortex, lacunes are rare but amyloid angiopathy is common. The aim of the present study is to compare the structure of perivascular spaces in the basal ganglia and at the anterior perforated substance with perivascular spaces in the cerebral cortex. Eight postmortem brains from patients aged 23-80 years (mean 68 y) were examined by light microscopy, by scanning and transmission electron microscopy and by direct visualisation of etched paraffin blocks. The results show that arteries in the basal ganglia are surrounded by 2 distinct coats of leptomeninges separated by a perivascular space which is continuous with the perivascular space around arteries in the subarachnoid space. The inner layer of leptomeninges closely invests the adventitia of the vessel wall and the outer layer is continuous with the pia mater on the surface of the brain at the anterior perforated substance. Veins in the basal ganglia have no outer layer of leptomeninges and thus the perivascular space is continuous with the subpial space. The anatomy of the periarterial spaces in the basal ganglia differs significantly from that in the cerebral cortex where there is only a single periarterial layer of leptomeninges. Differences in structure of perivascular spaces around arteries may reflect relative efficiencies in the drainage of interstitial fluid from different sites in the brain. Furthermore, the structure of the perivascular spaces may contribute to the relatively high frequency of lacunes in the basal ganglia, and the low frequency of amyloid angiopathy at this site in Alzheimer's disease. PMID- 9418991 TI - Total number and size distribution of motor neurons in the spinal cord of normal and EMC-virus infected mice--a stereological study. AB - The encephalomyocarditis virus of the diabetogenic M-strain (EMC-M) is known to cause diabetes in mice. The EMC-M virus has also been shown to cause paresis in some of the infected animals. The clinical features include an acute ascending predominantly motor paralysis, developing within days. This resembles acute idiopathic polyneuritis. The alpha motor neurons would be a possible target for the virus, so two parameters, the total number and the size distribution of motor neurons, were therefore selected for further investigation in 6 mice with neurological involvement and compared with 6 control mice. The optical fractionator method was applied for estimating the total number of motor neurons and the 3D size distribution was estimated using the rotator method in a vertical design. No difference was found in the total number of motor neurons and the size distributions were similar in the 2 groups. This design can be used as a model for the estimation of the total number of motor neurons and their size distribution in other experimental animal models. PMID- 9418989 TI - Animal models for inherited peripheral neuropathies. AB - Recent progress in human genetics and neurobiology has led to the identification of various mutations in particular myelin genes as the cause for many of the known inherited demyelinating peripheral neuropathies. Mutations in 3 distinct myelin genes, PMP22, P0, and connexin 32 cause the 3 major demyelinating subtypes of Charcot-Marie-Tooth (CMT) disease, CMT1A, CMT1B and CMTX, respectively. In addition, a reduction in the gene dosage of PMP22 causes hereditary neuropathy with liability to pressure palsies (HNPP), while particular point mutations in PMP22 and P0 cause the severe Dejerine-Sottas (DS) neuropathy. A series of spontaneous and genetically engineered rodent mutants for genes for the above mentioned myelin constituents are now available and their suitability to serve as models for these still untreatable diseases is an issue of particular interest. The spontaneous mutants Trembler-J and Trembler, with point mutations in PMP22, reflect some of the pathological alterations seen in CMT1A and DS patients, respectively. Furthermore, engineered mutants that either over or underexpress particular myelin genes are suitable models for patients who are similarly compromised in the gene dosage of the corresponding genes. In addition, engineered mutants heterozygously or homozygously deficient in the myelin component P0 show the pathology of distinct CMT1B and DS patients, respectively, while Cx32 deficient mice develop pathological abnormalities similar to those of CMTX patients. Mutants that mimic human peripheral neuropathies might allow the development of strategies to alleviate the symptoms of the diseases, and help to define environmental risk factors for aggravation of the disease. In addition, such mutants might be instrumental in the development of strategies to cure the diseases by gene therapy. PMID- 9418992 TI - Modulation of dye-coupling and proliferation in cultured rat thymic epithelium by factors involved in thymulin secretion. AB - Cultures of rat thymic epithelium were used to measure the effect of thymulin secretagogues on dye-coupling and proliferation. Dye-coupling was assessed after the injection of lucifer yellow dextran which cannot permeate the connexin pore of gap junctions and the smaller, permeant cascade blue. In addition to gap junctional communication, larger intercellular bridges were demonstrated by the transfer of lucifer yellow dextran between cells. The extent of intercellular communication was found to be influenced by both cell density and the number of passages. In control cultures, intercellular communication was reduced in cell groups of low (< 20 cells/group) or high cell densities (> 100 cells/group) compared with groups of 20-60 cells. The highest coupling indices were found in subcultures 20-30. Taking these factors into account, significant decreases in coupling index were observed after pretreatment of test cultures with factors known to influence the secretion of thymulin (5 U/ml interleukin 1 (alpha and beta), 1 microM progesterone, 1 microM oestrogen, 1 microM testosterone, 1 ng/ml adrenocorticotropic hormone, 100 nM rat growth hormone) but 7.5 ng/ml thymulin had no effect on dye-coupling. The nonspecific gap junction uncoupler, octanol, abolished dye-coupling. Cellular proliferation, as measured by the uptake of tritiated thymidine, showed that the same factors that reduced coupling also increased proliferation. None of these factors affected the number of multinucleate cells present, except interleukin-1beta which caused a significant reduction in the average number of nuclei per cell. Thus rat thymic epithelium in vitro provides a model for the study of the direct action of factors on cells of the thymic microenvironment. PMID- 9418993 TI - Immunohistochemical methods for semiquantitative analysis of collagen content in human peripheral nerve. AB - Methods are described for the semiquantitative analysis of the connective tissue components of human peripheral nerve using light microscopy. General histological preservation was assessed using haematoxylin and eosin staining and the distribution of collagen type IV was investigated using immunohistochemistry. Several techniques were investigated to establish the one giving optimum structural preservation, immunobinding and greatest contrast for image analysis. Frozen sections were unsuitable for this tissue and paraffin wax sections were therefore used. Alcohol fixation was rejected due to poor preservation of the endoneurium, although immunobinding was excellent. Ice-cold formalin fixation for 24 h was found to be adequate for structural preservation and antibody binding, provided that a protease step was introduced. Trypsin was found to be superior to pepsin for exposing collagen type IV epitopes. Of the detection systems investigated indirect immunofluorescence was not suitable due to considerable autofluorescence of the nerve. The avidin-biotin method provided the greatest contrast, and was therefore the detection method of choice for image analysis. The optimum techniques for image analysis were then used on control human sural nerve to ascertain the best comparative method for collagen type IV in the perineurium. A method of semiquantitative analysis is described which takes into account the fact that there is a close linear relationship between collagen content per unit of perineurium and perineurial perimeter as fascicle size increases in peripheral nerve. This means that data from 2 different sample groups can easily be compared, provided that a range of fascicle sizes is analysed in each case. PMID- 9418994 TI - Morphometric study of glycine-immunoreactive neurons and terminals in the rat cuneate nucleus. AB - The distribution of glycine-immunoreactive (glycine-IR) neurons and their associated axon terminals in the rat cuneate nucleus was studied using antiglycine postembedding immunoperoxidase labelling and immunogold staining, respectively. The immunoperoxidase-labelled glycine-IR neurons were widely distributed in the entire rostrocaudal extent of the nucleus. They made up 30.8% (9671/31368) of the neurons surveyed. Quantitative evaluation showed that the percentage of glycine-IR neurons in the caudal level was significantly higher than that in the middle and rostral levels. The glycine-IR neurons were small cells (mean area = 198+/-1.9 microm2, n = 2862) with ovoid or spindle-shaped somata. Statistical analysis showed that the size of the glycine-IR neurons in the rostral level was significantly smaller than that in the middle and caudal levels. Immunogold labelled glycine-IR terminals which contained predominantly pleomorphic synaptic vesicles were mostly small (mean area = 1.24+/-0.03 microm2, n = 286) and they constituted 24.7% (286/1158) of the total terminals surveyed. They formed axodendritic, axosomatic and axoaxonic synapses with unlabelled elements. It is suggested from this study that glycine is one of the major neurotransmitters involved in the depression of synaptic transmission in the cuneate nucleus. PMID- 9418995 TI - Comparative morphology of the alimentary tract and its glandular derivatives of captive bustards. AB - This study describes the gross anatomy of the alimentary tract of Houbara Bustards (Chlamydotis undulata macqueenii), Kori Bustards (Ardeotis kori), Rufous crested Bustards (Eupodotis ruficrista) and White-bellied Bustards (Eupodotis senegalensis) maintained in captivity by the National Avian Research Center in the United Arab Emirates. The morphology of the alimentary tract and the proportions of each region were similar in all 4 species. The length of the oesophagus, combined proventriculus and ventriculus, small intestine, and large intestine formed 24.2-28.4%, 7.3-9.7%, 40.5-55.1% and 9.1-14.7% of the total alimentary tract length respectively. Neither crop nor oesophageal enlargement was observed in the birds examined in this study, although male Kori Bustards possessed a saccus oralis in the oropharyngeal cavity. Oesophagi, proventriculi, ventriculi, caeca and large intestine were well developed in all species. The small intestine was shorter than that of other avian herbivores and granivores when compared on a bodyweight basis. The well differentiated stomachs and well developed caeca of the bustards examined in this study are characteristic of omnivores. Analysis of the mean lengths of the alimentary tract components and weight of the liver and pancreas showed sexual dimorphism in cases where male and female data were available for direct comparison. PMID- 9418996 TI - Adaptation of the disector method to rare small organelles in TEM sections exemplified by counting synaptic bodies in the rat pineal gland. AB - The disector is the only objective method for quantifying particles of variable size in a given volume. With this method, cell organelles are identified on adjacent sections, but only those present in one section are counted. When counting extremely rare structures in transmission electron microscope sections (physical disector), the usual procedure of counting on electron micrographs is limited for economic reasons (e.g. micrographs highly outnumbering the investigated structures). Hence, to apply this unbiased stereological method, a modification of the physical disector concerning 3 aspects has been developed. (1) The prerequisite of screening large corresponding tissue areas (here approximately 65000 microm2) was fulfilled by examining tissue areas along the edges of ultrathin sections. (2) The size of the counting frame was determined by measuring the lengths of the section margins (minus a guard area) by means of a Morphomat. This value was multiplied by the width of the investigated tissue zone, corresponding to the diameter of the electron microscope viewing screen. (3) Disector counting was carried out simultaneously on both sections (bidirectional disector) to improve efficiency. In the present study tiny synaptic bodies (SBs) were quantitated by disector in a rat pineal gland, yielding approximately 30 SBs/1000 microm3. By contrast, single section profile counts of SBs amounted to 90 SBs/20000 microm2. Since the presently described adaptation of the disector is time-consuming, it is proposed to determine a proportion factor allowing to estimate number of structures per volume based on single section profile counts. This would decrease the evaluation time by more than 50%. PMID- 9418997 TI - Age-related changes in cortical porosity of the midshaft of the human femur. AB - Complete cross-sections from the femoral midshaft of 180 individuals of known height and weight, aged 21-97 y, from a modern Australian population were examined using automatic video image analysis to quantify total subperiosteal porosity (TSPP). More specifically, the aim was to investigate whether age changes were similar in both sexes in (1) total subperiosteal area (TSPA), cortical area (CA) and medullary area (MA), (2) intracortical porosity (ICP), and (3) the respective contributions to TSPP made by MA and intracortical void area (ICVA). Our findings indicated that both sexes showed a significantly greater height normalised TSPA in the 70s as compared with the 20s. Males had consistently larger bones with a greater height normalised CA. In both sexes CA showed a tendency to increase till the 7th decade and then to decline, more so in females. MA approximately trebled in females and doubled in males over the age range studied. Although ICP also increased, from 4-6% in young adults to over 9% in the elderly, it showed a significant difference between the sexes only in the 3rd decade, being greater in males at this stage. By contrast, TSPP became significantly greater in females (from that recorded in the 3rd decade) by the time they reached the 50s, while in males this did not occur till the 80s. It increased from approximately 25% in young adults of both sexes to approximately 50% in females and approximately 37% in males in their 80s. However, in the elderly there was great variability in both sexes in the appearance of bones from individuals of similar chronological age. Some bones differed little from those in younger subjects, others showed greatly increased ICP, still others displayed reduced cortical widths with low ICP. The femoral midshaft resembles other skeletal sites in that age changes in TSPP are more marked in females than males. PMID- 9418998 TI - Distribution of myocardial macrophages in the normal human heart. AB - Macrophages are important in inflammatory processes in heart disease and in transplantation rejection. A resurgence of interest in the macrophage has emanated from recent evidence implicating it as an effector cell in atherosclerosis and transplantation rejection. The detailed distribution of the macrophage within the normal human heart is unknown. We quantified macrophage numbers in the different chambers of the heart. Large tissue blocks (1.5-2.0 cm3) were removed from specific sites in 5 'normal' control hearts (2 males, 3 females, age range 19-46 y). Paraffin-embedded sections were stained with a CD68 pan macrophage marker. Positive cells were enumerated within 20 random fields. Results were analysed using a generalised linear modelling method using the Poisson distribution. Macrophages were identified within septa, and often close to blood vessels, in the myocardium, and in the majority of areas in all hearts. Macrophage numbers varied significantly between areas (range 0-6 cells/high power field; P < 0.001), and between the 5 hearts analysed (P < 0.001). In general, there were significantly more macrophages in the ventricles (RV P < 0.01, LV P < 0.05), but these differences were affected by heart differences. This study provides a baseline for the range of macrophage numbers within normal hearts, thus enabling comparisons with macrophage numbers within diseased and transplanted hearts. PMID- 9418999 TI - Proliferative activity of preovulatory follicles and newly formed corpora lutea in cycling rats from late prooestrus to early oestrus. AB - Ovaries from adult cycling rats were studied from 1600 h on the day of prooestrus to 0700 h on the day of oestrus in order to relate the cyclic hormonal changes to the proliferative activity of preovulatory and postovulatory (i.e. newly-formed corpora lutea) follicles. Proliferative activity was studied by the immunohistochemical demonstration of DNA-incorporated 5-bromodeoxyuridine (BrdU). The proliferative activity of granulosa cells (GC) in large preovulatory follicles showed a centripetal pattern and decreased during prooestrus, reaching a minimum at 2100 h. However, a proliferative wave was found in the GC of preovulatory follicles at 0200 h on the day of oestrus and in those of newly formed corpora lutea at 0700 h on the day of oestrus. These results suggest that the granulosa cells of preovulatory follicles show maturational changes that followed a different pattern, depending on their location within the follicle, and that the proliferative wave found from 0200 to 0700 h on oestrus is important for the establishment of the number of steroidogenic cells in the cyclic corpus luteum. PMID- 9419000 TI - Colocalisation of insulin and IGF-1 receptors in cultured rat sensory and sympathetic ganglion cells. AB - Peripheral sensory and autonomic neurons are known to possess insulin receptors. These have been considered to be of the peripheral type, i.e. similar to those of hepatic and fat cells rather than of the brain type which show dual specificity for both insulin and insulin-like growth factor (IGF-1). We have examined the localisation of insulin and IGF-1 receptors in cultured sensory and sympathetic ganglion cells using confocal microscopy and indirect labelling with FITC (fluorescein isothiocyanate) and TRITC (tetramethyl rhodamine isothiocyanate) respectively. We have shown that in cultured U266B1 multiple myeloma cells these receptors display separate localisation, whereas they are colocalised in IM-9 lymphocytes which are known to possess hybrid receptors. We have confirmed the sequestration of insulin and IGF-1 receptors in the cytoplasm of sensory and sympathetic neurons, consistent with a brain-type receptor. The colocalisation of insulin and IGF-1 receptors in sensory and sympathetic ganglion cells is consistent with the view that they are hybrid receptors, similar to those present in the CNS. The function of these receptors, as suggested for the CNS, may be related to trophic support for neurons. PMID- 9419001 TI - Studies on rat and human thymus to demonstrate immunoreactivity of calcitonin gene-related peptide, tyrosine hydroxylase and neuropeptide Y. AB - The peptidergic and noradrenergic innervation of rat and human thymus was investigated by immunohistochemistry at the light and electron microscopical level (avidin-biotin-complex, sucrose-phosphate-glyoxylic-acid, and immunogold techniques). The distribution of noradrenergic neural profiles, and positive immunoreactivity for calcitonin gene-related peptide (CGRP), tyrosine hydroxylase (TH) and neuropeptide Y (NPY) is described in female rats during ageing, and in human children. In the neonatal rat thymus, the arteries and septa are well supplied by fine varicose nerves. In older animals (2 wk-1 y) the number of septa and blood vessels increase and consequently also the innervation. No nerves were found in the cortex. Apart from the innervation of the septal areas, immunoreactivity for CGRP and TH was present in thymic cells. Except for the young rats (neonatal-14 d), all rats showed CGRP positivity in subcapsular/perivascular epithelial cells (type 1 cells). All rat thymuses also contained a few TH positive cells in the medulla, which could only be confirmed as epithelial cells (type 6 cells) in children. Type 1 cells in the human thymus were not CGRP positive, but as in the rat, there were similar TH positive cells in the medulla. It was concluded that in addition to nerves containing CGRP, noradrenaline or dopamine, epithelial cells also contain these transmitters. They could therefore act on different cells (compared with neural targets) in a paracrine manner. PMID- 9419002 TI - Incidence and morphology of accessory heads of flexor pollicis longus and flexor digitorum profundus (Gantzer's muscles) PMID- 9419003 TI - Anatomical variations of the anterior talofibular ligament of the human ankle joint. PMID- 9419004 TI - Absence of the musculocutaneous nerve with innervation of coracobrachialis, biceps brachii, brachialis and the lateral border of the forearm by branches from the lateral cord of the brachial plexus. PMID- 9419005 TI - The iliopubic groove: a possible consequence of incomplete triradiate fusion. Two case reports. PMID- 9419006 TI - An unusually distal and complete additional flexor profundus muscle to the index finger. PMID- 9419008 TI - The glenoid notch and the shape of the glenoid cavity of the scapula. PMID- 9419007 TI - A shortened and deformed humerus from early modern Lithuania (16th/17th century A.D.): an unusual case of amputation in childhood? PMID- 9419009 TI - Schwann cells modulate sodium channel expression in spinal sensory neurons in vitro. AB - In order to study the factors that govern the expression of sodium channel alpha , beta1- and beta2-subunits, the influence that Schwann cells (SC) exert in the expression of sodium channels in DRG neurons was examined with in situ hybridization, immunocytochemistry, and patch clamp recording. The expression of sodium channel alpha-, beta1-, and beta2-subunit mRNAs in DRG neurons isolated from E15 rats cultured in defined medium in the absence (control) or presence of SC, or in SC-conditioned medium, was examined with isoform-specific riboprobes for sodium channel alpha-subunits I, II, III, NaG, Na6, hNE/PN1, SNS, and beta1- and beta2-subunits. DRG neurons cultured in the presence of SC displayed a significant (P < 0.05) increase in the hybridization signal for NaG, Na6, SNS, and Na beta2 mRNAs in comparison to control DRG neurons. In contrast, in SC conditioned medium, only the hybridization signal for SNS mRNA was significantly increased. The upregulation of sodium channel mRNAs in DRG neurons co-cultured with SC was paralleled by an increase in sodium channel immunoreactivity of these cells. An increase in the mean sodium current density in DRG neurons in the presence of SC was also observed. These results demonstrate that a SC-derived factor selectively upregulates sodium channel alpha- and beta-subunit mRNAs in DRG neurons isolated from E15 rats that is reflected in an increase in functional sodium channels in these cells. This culture system may allow elucidation of the SC factor(s) that modulate the expression of sodium channels in DRG neurons. PMID- 9419010 TI - Expression of alpha vbeta3 and alpha vbeta8 integrins during oligodendrocyte precursor differentiation in the presence and absence of axons. AB - We have shown previously that switching of the alpha v-associated beta1 and beta5 integrin subunits during differentiation of myelin-forming oligodendrocytes may regulate important aspects of cell behaviour such as migration (Milner et al., 1996: J Neurosci 16:7240-7252). In this study we have examined the developmental regulation of other alpha v-associated beta subunits in oligodendroglial cell cultures and also the control of their expression by neurons, using xenocultures to distinguish glial and neuronal integrins. We have found that oligodendroglia express alpha vbeta8 in addition to the previously-described alpha vbeta1, alpha vbeta3, and alpha vbeta5. Beta8 and beta3 together comprise the 80kD band seen in alpha v immunoprecipitations that represents the most abundant alpha v-associated beta subunit and show reciprocal patterns of expression during development. Alpha vbeta8 is expressed at high levels on oligodendrocyte precursors and differentiated oligodendrocytes but diminishes during the intermediate stages of differentiation. Alpha vbeta3, in contrast, shows an opposite pattern of expression, with the highest levels seen at the intermediate stages of differentiation and little expression on either oligodendrocyte precursors and differentiated oligodendrocytes. The expression of alpha vbeta3 is not altered by coculture with neurons, unlike that of alpha vbeta8, in which the decrease seen at the intermediate stages of differentiation is less marked in the presence of neurones. Our results confirm that switching of alpha v-associated beta subunits is an important feature of oligodendrocyte differentiation and suggest that alpha vbeta8 and alpha vbeta3 have distinct functions during myelination. PMID- 9419011 TI - Characteristics of thrombin-induced calcium signals in rat astrocytes. AB - The protease thrombin seems to play a central role in events following neural injury, whereby the enzyme can act, in concert with other molecules as a hormone or as a growth factor. In cells derived from the nervous system, thrombin induces changes in morphology and proliferation. The signalling mechanisms involved in these thrombin-activated processes are still unclear. In the present study we investigated Ca2+ signals in fura-2 loaded rat astrocytes in primary culture. Brief stimulation of astrocytes with thrombin induced a dose-dependent transient elevation of [Ca2+]i, best fitted by a double-sigmoidal curve giving two EC50 values of 3 pM and 150 pM. Continuous superfusion of cells with thrombin induced Ca2+ responses with three different types of kinetics. In 48% of the cells tested a single transient rise superimposed with fast fluctuations of [Ca2+]i was seen. The following complex long-term changes of [Ca2+]i, dependent on the presence of the agonist thrombin, were observed: i) a biphasic [Ca2+]i elevation, characterized by an initial peak followed by a sustained plateau phase (in 43% of the cells) and ii) oscillations of [Ca2+]i (in 9% of the cells). The observed Ca2+ responses were inhibited by the phospholipase C (PLC) inhibitor U-73122 and the thrombin inhibitor protease nexin-1/glia-derived nexin. The synthetic thrombin receptor activating peptide could mimic the thrombin-induced changes of [Ca2+]i. In astrocytes in Ca2+-free medium, thrombin induced a sharp single transient Ca2+ rise, without superimposed fluctuations. After depletion of intracellular Ca2+ stores with thapsigargin the Ca2+ response to thrombin was diminished or completely suppressed indicating that thrombin induces the release of Ca2+ from intracellular stores. During long-term Ca2+ responses, omission of extracellular Ca2+ resulted in a reversible interruption of the signal. In conclusion our results demonstrate that thrombin by activation of its plasma membrane receptor induces through activation of PLC different types of Ca2+ responses. The complex Ca2+ signals are generated by an interplay of InsP3 mediated Ca2+ release from intracellular stores and Ca2+ entry across the plasma membrane. PMID- 9419012 TI - Activated astrocytes induce nitric oxide synthase-2 in cerebral endothelium via tumor necrosis factor alpha. AB - Astrocytes under pathological conditions become activated and produce a variety of cytokines and low molecular weight signal molecules. Previously we demonstrated that activated astrocytes release nitric oxide which can downregulate the expression of nitric oxide synthase (NOS)-2 in co-cultured cerebral endothelium, and also release a transcriptionally regulated factor that can induce NOS-2 expression in endothelium (Borgerding and Murphy: J Neurochem 65:1342, 1995). The activity of this NOS-2-inducing factor was impeded by inhibitors of tyrosine kinases and NF-kappaB activation. Tumor necrosis factor (TNF alpha) alone, or in combination with IL-6, induced NOS-2 expression in endothelial cells. A neutralizing antibody against TNF alpha attenuated the NOS-2 expression in endothelial cells exposed to activated astrocytes. These results imply that cytokine-activated astrocytes release TNF alpha which can induce NOS-2 expression in endothelium and suggest that activated astrocytes within the CNS may induce expression of NOS-2 in cells of the adjacent microvasculature. The ensuing alterations in blood-brain barrier properties may be either beneficial or detrimental. PMID- 9419013 TI - Glutamate receptor-mediated calcium responses in acutely isolated hippocampal astrocytes. AB - GFAP(+) cells were acutely isolated from the hippocampi of 1-10 day old rats, and the intracellular calcium responses to L-glutamate, ATP, and 5-HT were studied in single cells. Eighty-two percent of such cells responded to glutamate, 20% of them responded to ATP, and none responded to 5-HT. The same cells that failed to respond to ATP and 5-HT often responded to glutamate. These proportions of cells responding to glutamate and ATP are very similar to those reported for GFAP(+) astrocytes in hippocampal slices (Porter and McCarthy, 1995a,b). After culturing for 1-2 days in serum-containing medium, 60% of such acutely isolated cells responded to either glutamate or ATP, and 5% to 5-HT. After 1 week in culture, the percentage of cells responding to glutamate remained essentially the same (62%) but the percentages of cells responding to ATP and 5-HT increased to 92% and 62%, respectively. These percentages were very close to the results obtained from primary hippocampal astrocyte cultures prepared from 1 day old rats and cultured for 1-2 weeks in vitro. Pharmacological characterization showed that the Ca2+ responses of acutely isolated hippocampal astrocytes from P1-10 rats was due to activation of a group I metabotropic glutamate receptor. The calcium responses to ATP and 5-HT in acutely isolated cells that were then cultured were mediated by P2y and 5-HT2A receptors, respectively. These data show that, like cortical astrocytes (Kimelberg et al., 1997), GFAP(+) astrocytes cultured from the hippocampi of young rats showed marked differences in receptor expression compared to their acutely isolated counterparts. Also, since the astrocytes acutely isolated from these 2 different brain regions showed qualitatively the same responses for the 3 receptors selected, it indicates a degree of homogeneity of receptor expression for astrocytes from these 2 brain regions. PMID- 9419014 TI - Opposite influence of the metabotropic glutamate receptor subtypes mGlu3 and -5 on astrocyte proliferation in culture. AB - In non-synchronized, subconfluent secondary cultures of rat cortical astrocytes, the selective group-I metabotropic glutamate (mGlu) receptor agonist 3,5 dihydroxyphenylglycine (DHPG) increased [methyl-3H]-thymidine incorporation. This effect was mediated by the activation of the mGlu5 receptor, which was shown to be present by either RT-PCR or Western blot analysis. The mixed mGlu receptor antagonist (+)-alpha-methyl-4-carboxyphenylglycine reduced the increase in both intracellular Ca2+ and [methyl-3H]-thymidine incorporation produced by DHPG. In contrast, (2S,1'R,2'R,3'R)-2-(2,3-dicarboxycyclopropyl)glycine (DCG-IV), a potent and selective agonist of group-II mGlu receptors, reduced [methyl-3H]-thymidine incorporation in non-synchronized astrocyte cultures. The antiproliferative effect of DCG-IV was prevented by the selective group-II mGlu receptor antagonist (2S,1'S,2'S,3'R)-2-(2'-carboxy-3'-phenylcyclopropyl)glycine (PCCG-IV). The opposite effect of DHPG and DCG-IV on astrocyte proliferation was confirmed in cultures deprived of serum for 48 hours and then stimulated to proliferate with either epidermal growth factor (EGF) or the metabolically stable ATP analogue adenosine 5'-(beta,gamma-imido)-triphosphate (AMP-PNP). We conclude that activation of mGlu5 receptors enhances proliferation in cultured astrocytes, whereas activation of a receptor with pharmacological characteristics similar to those of mGlu2/3 receptors reduces proliferation. PMID- 9419016 TI - Modulation of T cell-endothelial adhesion by astrocyte conditioned medium. AB - Astrocytes and derived factors maintain the morphologic, phenotypic, and physiological properties of the blood-brain barrier. Astroglial cells may also modulate endothelial cell properties associated with the entry of inflammatory cells into the brain. The study of mechanisms of lymphocyte migration through the blood-brain barrier is critical to understanding the pathophysiology of autoimmune (multiple sclerosis) and virus-induced central nervous system diseases (HIV-induced dementia). In this context the contribution of astrocyte derived factors in regulating the interactions between inflammatory cells and endothelial cells of the blood-brain barrier was studied. The treatment of endothelial cells derived from brain or peripheral sources (hepatic) with astrocyte conditioned medium resulted in a dose dependent enhancement of adhesion of T cells to endothelium. The antigen specificity of the T cells did not influence the findings. Identical results were obtained with fresh Concanavalin A activated T cells and T cell hybridomas generated using myelin basic protein or chicken ovalbumin as immunogens. Further studies are in progress to define the active components in astrocyte conditioned medium and endothelial cell adhesion molecules that are regulated in order to gain a better understanding of mechanisms of inflammatory cell entry into the central nervous system. PMID- 9419015 TI - A nestin-negative precursor cell from the adult mouse brain gives rise to neurons and glia. AB - Using a novel suspension culture approach, previously undescribed populations of neural precursor cells have been isolated from the adult mouse brain. Recent studies have shown that neuronal and glial precursor cells proliferate within the subependymal zone of the lateral ventricle throughout life, and a persistent expression of developmentally regulated surface and extracellular matrix molecules implicates cell-cell and cell-substrate interactions in the proliferation, migration, and differentiation of these cells. By using reagents that may affect cell-cell interactions, dissociated adult brain yields two types of cell aggregates, type I and type II spheres. Both sphere types are proliferative, and type I spheres evolve into type II spheres. Neurons and glia arise from presumptive stem cells of type II spheres, and they can survive transplantation to the adult brain. PMID- 9419017 TI - Albumin enhances superoxide production in cultured microglia. AB - Microglial activation and disruption of blood-brain barrier (BBB) are known to occur and contribute to neuronal damages in cerebral ischemic conditions and in some neurodegenerative diseases. To investigate whether a serum factor leaked out from circulation enhances microglial activation, we examined the effect of normal rat serum on superoxide (O2-) production by cultured microglia. Microglia cultured from neonatal rat brains were studied on their O2- production induced by the addition of phorbol myristate acetate by a method of acetyl-cytochrome c reduction. The O2- production was significantly increased by the addition of 0.01% rat serum, and the maximal enhancement was observed at about 0.1% rat serum. After the serum was fractionated using a molecular sieve membrane, we observed the enhancing effect only in a greater molecular weight fraction (>50 kDa). Furthermore, three kinds of bovine serum albumin (BSA) with different purity, and human serum and plasma albumins, also enhanced O2- production to a similar extent to that by rat serum. However, other proteins tested showed no significant effect. The enhancement of O2- production by BSA was observed dose dependently, and the effect of 50 microg/ml of purified BSA was equivalent to that of 0.1% rat serum, suggesting that albumin itself enhances O2- production by microglia. These results imply that albumin leaked out through impaired BBB may activate microglia and that the potentiation of O2- production by albumin results in the pathogenesis of neuronal damage in cerebral ischemia and some neurodegenerative diseases. PMID- 9419018 TI - Effects of in-vivo generation of oxygen free radicals on immune responsiveness in rabbits. AB - Oxygen free radicals (OFRs) generated during biological processes are reportedly involved in the pathogenesis of several disease states and various reports have indicated that oxidative stress may alter immune competence. Hence, effects of in vivo generation of OFRs by using xanthine/xanthine oxidase (X/XO) system on immune responsiveness were evaluated in rabbits. Intravenous injections of xanthine (0.14 mg/kg) along with xanthine oxidase (2 U/Kg) following primary and secondary immunizations of animals with sheep red blood cells (SRBC) significantly attenuated the primary and secondary antibody responses respectively. In tests for cell-mediated immunity, tuberculin sensitivity and leucocyte migration inhibition were also decreased significantly in sensitized animals following X/XO treatment. The observed changes in both humoral and cell mediated immune responses following such in-vivo generation of OFRs indicate a possible nexus between OFR generation and immune suppression. PMID- 9419019 TI - A FIV epitope defined by a phage peptide library screened with a monoclonal anti FIV antibody. AB - Phage peptide libraries constitute powerful tools for the mapping of epitopes recognized by monoclonal antibodies. We report here the characterization of an antibody directed against a 20-residue peptide derived from the surface glycoprotein of the feline immunodeficiency virus. The isolation of the WRPDF consensus sequence from a phage display library defined the exact epitope recognized by the mAb. Compared with known immunogenic peptides of the FIV envelope, it corresponds to the most immunodominant peptide found in the whole molecule. Kinetic data describing the antibody-peptide interactions were obtained by surface plasmon resonance. The antibody binds the peptide with a KD in the nanomolar range. PMID- 9419020 TI - Tumor necrosis factor alpha (TNF-alpha) activates human adenoidal and cutaneous mast cells to histamine secretion. AB - It is widely known that mast cells and cytokine TNF-alpha are both involved in inflammatory reactions. Therefore, we have studied whether TNF-alpha can cause histamine secretion from human adenoidal and cutaneous mast cells. The experiments were performed in vitro on mast cells isolated from tissues by enzymatic dispersion technique. The results of our experiments have clearly shown that this cytokine stimulates mast cells to histamine release. TNF-alpha-induced histamine release was concentration- and time-dependent. Moreover, the release of histamine evoked by TNF-alpha was also dependent on reaction temperature and on glycolytic and oxidative cellular metabolism. We have concluded that TNF-alpha is a potent stimulus for mast cells to release histamine and that it induces histamine release via an active, secretory process. PMID- 9419021 TI - Topical immune modulation (TIM): a novel approach to the immunotherapy of systemic disease. AB - In this article, we present the concept of topical immune modulation, or TIM. TIM is based on the observation that skin contact sensitizing agents such as poison ivy, poison oak and dinitrochlorobenzene (DNCB) are potent stimulants of the cellular immune system that combats viruses and other pathogens. We discuss the evolution of DNCB as a therapeutic modality in the acquired immunodeficiency syndrome (AIDS) and we explore the mechanism by which DNCB directs the immune response. The potential use of topical immune modulators in autoimmune disease and vaccine development is also delineated. TIM represents a novel approach to immunotherapy that should have widespread application for immunologic diseases. PMID- 9419022 TI - The 9-O-acetylated disialosyl carbohydrate sequence of CDw60 is a marker on activated human B lymphocytes. AB - Gangliosides with a terminal 9-O-acetylated disialosyl group (CDw60 structures) show a restricted surface expression on human leukocytes. Hithereto, they have only been detected on subpopulations of human T lymphocytes. Using the defined CDw60 antibody UM4D4 and two new antibodies with preferential CDw60 activities, F6 and Z17, we demonstrate for the first time that CDw60 is an activation marker on human B lymphocytes. In vitro phorbol ester-stimulated human peripheral blood B lymphocytes as well as in vivo activated tonsillar B lymphocytes became CDw60 positive. CDw60 expression of these cells exceeds that of resting and activated T lymphocytes. PMID- 9419023 TI - Induction of autoimmune-like hepatic and ductal lesions by administration of lipopolysaccharide in mice undergoing graft-versus-host reaction across MHC class I difference. AB - In this paper, we examined the induction of autoimmune-like histologic changes in the liver and other organs of mice undergoing graft-versus-host reaction (GVHR) with MHC class I disparity by the administration of bacterial lipopolysaccharide (LPS), on the assumption that stimulation with LPS could be an exacerbating factor. Spleen cells of C57BL/6 (B6) mice were injected twice into (B6 x bml) F1 recipient mice at an interval of 7 days to induce MHC class I GVHR and then challenged with 1 microg of LPS intravenously on the next day of the cell transfer. The hepatic lesions of the group of MHC class I GVHR mice challenged with LPS showed marked cellular infiltration at the portal area and focal necrosis was observed in the hepatic lobule. The major infiltrating cells were CD8+, and others including CD4+ cells being of minor populations. In addition, ductal lesions in extrahepatic organs, including the pancreas and salivary glands also showed marked cellular infiltration. Thus, we have demonstrated that LPS induced ductal lesions in mice with MHC class I disparity. CD8+ cells were detected at the destructive hepatic lesions, which might be effector cells. These findings indicate that LPS might be one of the potential factors which augment autoimmune-like lesions. PMID- 9419024 TI - Microheterogeneity of the cell surface tyrosine phosphatase, CD45RA, on T cells: phytohaemagglutinin binding and non-binding fraction of the 220 kDa isoform. AB - The CD45 antigen is a family of the transmembrane tyrosine phosphatases expressed on cells of haemopoietic origin. The molecules are distinguished by the different aminoacid sequence and glycosylation on the N terminus. Although all isoforms are heavily glycosylated and exert receptor like structures on the extracellular part, the role of the glycosylation in the possible receptor function and the ligand of the CD45 has not been determined yet. In this study we have examined the binding of phytohaemagglutinin (PHA) to the different isoforms and its relation to the phosphatase activity. Immunoblotting experiments revealed that the 220 kDa form of the CD45RA contained a PHA binding fraction when immunoprecipitated with CD45RA monoclonal antibody (mAb), while an isoform with identical molecular mass immunoprecipitated by anti-CD45 did not bind PHA. We concluded that the 220 kDa form was heterogeneous with respect to PHA binding. Functional data also confirmed this heterogeneity: previous extraction of the PHA binding proteins resulted in the elimination of all the phosphatase activity from CD45, while only a part of that was removed from CD45RA immunoprecipitates. PMID- 9419025 TI - Chromosomal location of the human immunoglobulin lambda variable 8 (IGLV8) gene family outside the major lambda locus on chromosome 22q11.2. AB - Physical mapping of the human immunoglobulin lambda locus (IGL) on chromosome 22q11 has shown the existence of at least 52 variable region gene segments. These Vlambda genes are associated with EcoRI fragments detectable in Southern blots of genomic DNA samples. The current physical map of the IGL locus includes a unique Vlambda8 gene (IGL8a, accession no. Z73650) in a 3.7 kb EcoRI fragment. However, our Southern blot-EcoRI-restriction fragment length polymorphism studies on the Brazilian population using a specific probe for the Vlambda8 gene (pVL8 probe) have revealed the presence of two additional fragments bearing Vlambda8 sequences (8.0 kilobase (kb) at 100% frequency and 6.0 kb at 10% frequency). We have used human/rodent somatic cell hybrid DNAs to locate these new Vlambda8 genes outside the major locus on chromosome 22q11.2. Polymerase chain reactions using specific primers for the IGLV8a gene on the somatic hybrid panel showed that chromosome 8 (besides 22q11) also comprises Vlambda8 sequences. This finding represents evidence for the dispersion of the human IGLV8 gene family outside the major locus (orphan genes). PMID- 9419026 TI - Some suggestions for writing better clinical papers. PMID- 9419027 TI - Spontaneous bacterial peritonitis in patients with ventriculoperitoneal shunts. AB - Spontaneous bacterial peritonitis (SBP) is an infection of the peritoneal fluid in the absence of an obvious intra-abdominal source. It is most commonly diagnosed in patients with cirrhotic ascites, although it has been described in other syndromes as well. The organisms most frequently cultured from the peritoneum are those of intestinal flora; however, there are cases which have all the features of SBP, but remain culture negative. This article discusses 7 cases of SBP in patients with ventriculoperitoneal shunts; a combination which has previously not been described. The most significant features of these cases include: a remote history of shunt revision (mean 3.4 years), and cultures consistent with normal intestinal flora. None had a history of recent abdominal surgery, gastrostomy or wire-impregnated catheters. Cerebrospinal fluid cultures are often negative, and when positive, suggest SBP with an ascending shunt infection. While SBP is clearly differentiated from pseudocyst of the abdomen, it may represent a point on the continuum of intra-abdominal processes in the shunted patient. The precise etiology of SBP is unclear. A number of suggested theories are reviewed. It is proposed that patients with shunts may be predisposed to develop SBP because spinal fluid can behave as an ascitic fluid even in the absence of a peritoneal accumulation. Recommendations for the recognition and management of SBP in the shunted patient are discussed in detail. PMID- 9419028 TI - A new model for tethered cord syndrome: a biochemical, electrophysiological, and electron microscopic study. AB - In order to investigate the pathophysiology of the tethered cord syndrome, a few experimental models have been developed and used previously. In this study, the authors present a new experimental model to investigate the biochemical, electrophysiological, and histopathological changes in the tethered spinal cord syndrome. A model was produced in guinea pigs using an application of cyanoacrylate to fixate the filum terminale and the surrounding tissue to the dorsal aspect of the sacrum following 5-gram stretching of the spinal cord. The experiments were performed on 40 animals divided into two groups. The responses to tethering were evaluated with hypoxanthine and lipid peroxidation, somatosensory and motor evoked potentials, and transmission electron microscope examination. The hypoxanthine and lipid peroxidation levels significantly increased, indicating an ischemic injury (p < 0.01). The average hypoxanthine level in the control group was 478.8 +/- 68.8 nmol/g wet tissue, while it was 651.2 +/- 71.5 nmol/g in the tethered cord group. The lipid peroxidation level in group I was 64.0 +/- 5.7 nmol/g wet tissue, whereas it was 84.0 +/- 4.7 nmol/g in group II. In the tethered cord group, the latencies of the somatosensory and motor evoked potentials significantly increased, and the amplitudes decreased. These changes indicated a defective conduction in the motor and sensorial nerve fibers. In the transmission electron microscopic examinations, besides the reversible changes like edema and destruction in the gray-white matter junction, irreversible changes like scarcity of neurofilaments and destruction in axons and damage in myelin sheaths were observed. We consider that this work can be used as an experimental model for tethered cord syndrome. PMID- 9419029 TI - Combined frameless stereotaxy and neuroendoscopy in placement of intracranial shunt catheters. AB - Twenty-six patients have had cranio-peritoneal shunts placed using a new introducer allowing the combination of frameless stereotaxy and neuroendoscopy and placement of a one-piece shunt. Operating times have been acceptable, complication rates have been low, and shunt placement has been accurate in this series. PMID- 9419031 TI - Use of positron emission tomography for presurgical localization of eloquent brain areas in children with seizures. AB - Successful surgical management of a neoplastic or nonneoplastic seizure focus in close proximity to or within eloquent brain areas relies on precise delineation of the relationship between the lesion and functional brain areas. The aim of this series was to validate the usefulness and test the efficacy of noninvasive presurgical PET mapping of eloquent brain areas to predict surgical morbidity and outcome in children with seizures. To identify eloquent brain areas in 15 children (6 female and 9 male; mean age 11 years) with epileptogenic lesions PET images of regional cerebral blood flow were performed following the administration of [(15)O]water during motor, visual, articulation, and receptive language tasks. These images with coregistered magnetic resonance (MR) images were then used to delineate the anatomic relationship of a seizure focus to eloquent brain areas. Additional PET images using [18F]fluoro-2-deoxy-D-glucose (FDG) and [11C]methionine (CMET) were acquired to help localize the seizure focus, as well as characterize the lesion. Patient surgical management decisions were based on PET mapping in combination with coregistered MR images, PET-FDG findings, and the anatomic characteristics of the lesion. At follow-up 1-26 months after surgery, all patients that underwent temporal lobectomy (9 patients) and extratemporal resection (4 patients) for a neoplastic or nonneoplastic seizure focus are seizure-free with minimal postoperative morbidity. Of prime importance, no child sustained a postoperative speech or language deficit. PET imaging was also well tolerated without procedural complications. Based on PET mapping, a nonoperative approach was used for 2 children and a biopsy only was used in one child. When cortical injury involved prenatally determined eloquent cortex, PET demonstrated reorganization of language areas to new adjacent areas or even to the contralateral hemisphere. Integration of anatomical and functional data enhanced the surgical safety, defined optimal surgical approach, delineated the seizure focus from eloquent brain areas, facilitated maximum resection and optimized the timing of surgery, thereby minimizing surgical morbidity while maximizing surgical goals. PET measurements of FDG and CMET uptake were also helpful in localizing the seizure focus and grading the tumors. PET used for brain mapping in children provides the surgeon with strategic preoperative information not readily attainable with traditional invasive Wada testing or intraoperative cortical stimulation. PET mapping may also improve the outcome of extratemporal resections by allowing aggressive seizure focus resection. In addition, serial brain maps may optimize timing for surgical intervention by demonstrating reorganization of eloquent cortex often seen in younger children after cortical injury. Our results suggest that noninvasive presurgical brain mapping has the potential to reduce risk and improve neurologic outcome. PMID- 9419030 TI - Progressive congenital kyphosis: report of five cases and review of the literature. AB - For over 60 years congenital kyphotic deformities of the spine have been categorized into two distinct groups, depending on the developmental defect. Those arising from a failure of formation of the vertebral bodies were classified as type 1, while those arising from a failure of segmentation were referred to as type 2. Recognition of the progressive and unstable nature of the type 1 defects alerted physicians to the need for early operative stabilization through decompression and stabilization through instrumentation. As the embryogenesis of the spinal column was further investigated, and as diagnostic imaging methods of the spine improved, unstable congenital kyphoses were further subdivided. Progressive congenital kyphotic deformities now may accompany a host of vertebral column developmental defects as well as genetically mediated mesenchymal tissue defect syndromes. This paper presents 5 patients from The Children's Hospital of Philadelphia with progressive and symptomatic congenital kyphotic deformities of the spine. Two of these lesions resulted from defects of formation of the vertebral bodies, while one resulted from segmental spinal dysgenesis, maldevelopment of both the anterior and posterior vertebral elements. One patient's kyphotic deformity was a result of caudal regression syndrome, and the final case presented experienced a high thoracic kyphosis from a syndrome associated diffuse midline mesenchymal tissue abnormalities known as cerebrocostomandibular syndrome. All patients showed evidence of progressive cord compression and required neural element decompression, fusion, and instrumentation. The cases are discussed individually, and the developmental and clinical aspects of each are explored. PMID- 9419032 TI - Fourth ventricular ependymoma in a child with Duane retraction syndrome. AB - A child with unilateral Duane retraction syndrome developed signs and symptoms of increased intracranial pressure at 3 1/2 years of age. Neuroimaging disclosed a fourth ventricular ependymoma compressing but not invading the floor of the fourth ventricle. The coexistence of Duane syndrome and fourth ventricular ependymoma in a young child raises the possibility that both conditions could have resulted from a mutational event or focus of cellular disorganization (i.e. field defect) localized to the dorsal pons. PMID- 9419033 TI - Spontaneous rapid resolution of an epidural hematoma associated with an overlying skull fracture and subgaleal hematoma in a 17-month-old child. AB - Acute traumatic epidural hematomas (EDH) constitute one of the most critical emergencies in neurosurgical management. The rapid spontaneous resolution (<24 h) of EDH is an extremely rare phenomenon. A 17-month-old patient fell from a height of 1.5 m and presented with a 8-mm temporal EDH, an overlying linear skull fracture, and a subgaleal hematoma without evidence of intraparenchymal injury or edema. The patient complained only of mild headache, harbored no neurological deficits, and was, therefore, managed conservatively in the intensive care unit with provision to proceed to surgical decompression in the event of neurological change. A repeat CT study 18 h later revealed near-complete resolution of the EDH with a coincident increase in the volume and spread of the subgaleal hematoma. This is the fourth reported case of rapid spontaneously resolving EDH and the youngest one to date. All 4 cases have coincided with an overlying linear skull fracture. We propose that unlike classical EDH, rapidly resolving EDH in the absence of elevated intracranial pressure (ICP) originates from elevated interstitial pressure in the subgaleal compartment transiently decompressing into the epidural space through a skull fracture and resolving as the pressure in the subgaleal compartment decreases below ICP. PMID- 9419034 TI - Images in pediatric neurosurgery. 3rd ventricle in the Chiari II malformation. PMID- 9419035 TI - Presentation and management of hydromyelia in children with Chiari type-II malformation. AB - Hydromyelia in patients with myelomeningocele and Chiari-II malformation is a relatively frequent finding on MRI studies. However, not all children develop symptoms from the hydromyelia that requires treatment. Furthermore, treatment of hydromyelia in spina bifida patients is rather complex due to the associated malformations. The authors retrospectively analyzed 231 MRI studies carried out on spina bifida patients who presented neurological deterioration. Hydromyelia was found in 48.5% of the patients. Forty-five children with severe hydromyelia required treatment. These patients were first divided into 2 groups: those with holocord hydromyelia, and those with a segmental lesion. Fifteen patients presented symptoms characteristic of symptomatic Chiari-II malformation: neck rigidity; swallowing difficulty; pain in the upper extremeties; weakness or spasticity in the upper extremeties. Eighteen patients presented symptoms typical of the tethered cord syndrome: scoliosis; worsening bladder and/or bowel function; pain in the lower extremeties; weakness or spasticity in the lower extremeties. Twelve patients presented a mixed-type symptomatology. These patients subsequently underwent posterior cervical decompression, tethered cord release or insertion of a hydromyelia-pleural shunt according to the type of presenting symptoms and to the extent of the hydromyelic lesion. A pattern of successful treatment was identified for each type of presenting clinical and radiological picture. This has allowed the authors to determine an algorithm for optimal treatment of hydromyelia associated with Chiari-II malformation and myelomeningocele, which is proposed here. PMID- 9419036 TI - Combined utility of functional MRI, cortical mapping, and frameless stereotaxy in the resection of lesions in eloquent areas of brain in children. AB - We studied 16 children with lesions in the eloquent brain to determine if the amalgamation of information from functional magnetic resonance imaging (fMRI), frameless stereotaxy, and direct cortical mapping and recording could facilitate the excision of these lesions while minimizing potential neurological deficits. The mean age of the children was 10 years. Fourteen children presented with seizures. All lesions were located in or near eloquent cerebral cortex. fMRI was successful in all patients in delineating the relationship between the lesion and regions of task-activated cortex. The ISG wand was utilized in all cases for scalp and bone flap placement, and for intraoperative localization of the lesion. Direct cortical stimulation or recording of phase reversals with somatosensory evoked potentials helped delineate the central sulcus and language cortex in patients with lesions near the motor or language cortex. Intraoperative electrocorticography (ECoG) was utilized in all patients who presented with seizures to guide the extent of resection of the epileptiform cortex. Ten children had benign cerebral neoplasms, nine of which were totally resected. The other diagnoses included vascular malformations, Sturge-Weber, tuberous sclerosis, Rasmussen's encephalitis, and primitive neuroectodermal tumor. Only 1 patient with a left Rolandic AVM developed a new neurological deficit postoperatively. Thirteen of fourteen patients who presented with seizure disorders were rendered either seizure free or improved in terms of seizure control postoperatively. Follow-up has ranged from 12 to 18 months, with a mean follow-up of 15 months. We conclude that the techniques of fMRI, frameless stereotaxy, direct cortical stimulation and recording can be utilized in sequence to accurately localize intracerebral lesions in eloquent brain, and to reduce the morbidity of resecting these lesions in children. PMID- 9419037 TI - Surgical treatment of focal epilepsy in children: results in 37 patients. AB - This report concerns 37 children and teenagers operated upon for intractable seizures between 1990 and 1994. Follow-up is at least 3 years. Fourteen children underwent pure temporal lobe resections; 71% are seizure free, and 93% have a better than 90% decrease in seizure frequency. The presence of a lesion on magnetic resonance imaging, the side of the lesion, or the presence of abnormal pathology had no influence on the result of resection. 28% of the children who had extratemporal resections are seizure free, and 83% have a greater than 90% decrease in seizure frequency. There was a trend to better results in those with a lesion on magnetic resonance imaging. In the small group with temporal plus extratemporal foci, the results were poor with only 60% showing a greater than 90% reduction in seizure frequency. PMID- 9419038 TI - Anterior cerebral artery territory infarction in neurocysticercosis: evaluation by MR angiography and in vivo proton MR spectroscopy. AB - Vascular complications of neurocysticercosis are common, but frequently not recognized. Mostly these are in the form of an endarteritis involving the smaller basal vessels due to basal exudates. Large vessel territory infarction has been clearly documented in not more than 10 cases. All these cases had either a chronic meningeal state or close proximity of cysts to the vessel wall explaining the vasculitis. We report a case that developed anterior as well as segmental middle cerebral artery territory infarction in the acute encephalitic state of neurocysticercosis. MR angiography showed constriction in the proximal segment of the right anterior cerebral artery. There was no biochemical or imaging evidence of a meningeal reaction. In vivo MR spectroscopy over the infarction showed absent N-acetyl-aspartate, low creatine and high lactate. This is the first case showing a large vessel territory infarction in the encephalitic state of neurocysticercosis, with no meningeal reaction. Focal arteritis due to an adjacent brain parenchymal reaction could be a possible mechanism for the vasculitis. MR spectroscopy may have a potential role in assessing tissue viability and therapeutic modalities in such infective vasculitis. PMID- 9419039 TI - Long-term survival of an infant with 'anaplastic' astrocytoma. AB - Anaplastic astrocytomas are intermediate in differentiation between astrocytoma and glioblastoma multiforme. Survival with anaplastic astrocytoma is favorably affected by extensive anaplasia, maximal resection and presentation in early life. We report a 2-month-old infant who had a tumor of astrocytic lineage with anaplastic features of necrosis, nuclear atypia and mitotic activity. Following subtotal resection the child is alive but has a radiologically visible tumor. PMID- 9419040 TI - Cerebral sparganosis in a child. AB - Cerebral sparganosis in childhood is very rare. Until 1996, 2 cases in children up to 15 years of age had been described. We report a case of cerebral sparganosis in a 6-year-old girl who presented with seizures. Sequential brain magnetic resonance imaging scans spaced over 4 months showed a lesion which migrated from the right parieto-occipital region to the right occipital region. The enzyme-linked immunosorbent assay for antisparganum antibody was positive. She most likely contracted sparganosis from worm-infested spring-water. A live worm with surrounding granulomatous tissue was removed, and the enzyme-linked immunosorbent assay for antisparganum antibody converted to negative 12 months following surgery. In areas of endemic sparganosis, the possibility of cerebral sparganosis should be considered, even in a child, if the patient shows a migrating granulomatous lesion. PMID- 9419041 TI - Balamuthia mandrillaris meningoencephalitis presenting with acute hydrocephalus. AB - The leptomyxid amoeba Balamuthia mandrillaris, previously believed to be a harmless soil-inhabiting organism, is now known to be a rare but consistently lethal cause of meningoencephalitis in humans. We report a case of amebic meningoencephalitis caused by B. mandrillaris which presented as a febrile illness with acute hydrocephalus. PMID- 9419042 TI - Renal artery stenosis in aortoarteritis: spectrum of disease in children and adults. AB - Nonspecific aortoarteritis is the commonest cause of renovascular hypertension (RVH) accounting for 87% of the patients in the present study. We compared the clinical and radiographic features and outcome in children (n = 16) and adult (n = 24) patients with aortoarteritis. Children have a shorter duration of disease and present more commonly with constitutional symptoms. All the patients were hypertensive; however, malignant hypertension and hypertensive encephalopathy were more common in children. Abdominal bruit and asymmetry of pulses were present only in 75 and 35% of the patients, respectively. Asymmetric kidney size on ultrasound was present in 15 of 24 adults, whereas 9 of 16 children had equal sized kidneys. Captopril renography had a better sensitivity for detection of RVH in children (13 of 16 in children vs. 12 of 24 in adults showing positive results). On intra-arterial digital substraction angiography, abdominal aortic involvement was invariable, whereas the thoracic aorta was involved less frequently in both age groups. Angiographic scores for the severity of vascular involvement was significantly lesser in children (6.87+/-4.8) as compared to adults (11.32+/-4.5). Thirteen of the 15 children were found suitable for revascularization, whereas 12 of 24 adults were not considered for revascularization as their kidneys were small and contributed to less than 10% of total function. Six of the adult patients underwent nephrectomy for the control of blood pressure. Results of angioplasty were also better in children than adults. We conclude that children present earlier with less severe vascular disease and respond better to revascularization, as compared to adults. PMID- 9419043 TI - Noradrenergic blood pressure dysregulation and cytosolic calcium in primary hyperparathyroidism. AB - The purpose of this study was to characterize the changes in noradrenergic blood pressure regulation and the causes of this dysregulation in patients with histologically proven primary hyperparathyroidism before and after parathyroidectomy. In untreated hypertensive patients with primary hyperparathyroidism slightly higher plasma levels of norepinephrine, enhanced cardiovascular reactivity to norepinephrine (p<0.05) and increased cytosolic free calcium concentration in platelets (p<0.05) were found. Parathyroidectomy resulted in the correction of noradrenergic blood pressure dysregulation and in the reduction of cytosolic free calcium and led to normotension. In vitro incubation of platelets from normal subjects with heterologous plasma ultrafiltrates from either hypertensive patients with primary hyperparathyroidism or normotensive uremic patients with secondary hyperparathyroidism led to raised platelet calcium in primary hyperparathyroidism but not in secondary hyperparathyroidism, although in both conditions intact parathormone levels were elevated equally. Furthermore, when platelets of normotensive subjects were preincubated with commercially available parathormone, no effect on resting platelet cytosolic calcium was observed. Our results suggest that hypertension in primary hyperparathyroidism may be caused in part by noradrenergic blood pressure dysregulation and seems to be causally linked to the secretion of a parathyroid hypertensive factor, which increases cytosolic calcium and alters vascular tone and reactivity. PMID- 9419044 TI - Effect of PD 123319, an AT-2 antagonist, on renal function of the anesthetized dog: comparison with EXP 3174, an AT-1 blocker. AB - In anaesthetized dogs fed a diet delivering 3.5 mmol NaCl/kg/day, PD 123319 - an angiotensin (AT) II (Ang II) antagonist preferentially bound to AT-2 receptors - was infused into the renal artery for 20 min at 10 microg/kg/min, the lowest effective dose (group 1 dogs). In group 2 dogs, EXP 3174 (30 microg/kg/min) - an AT-1 blocker - was coinfused while, in group 3, EXP 3174 was infused alone. In all groups, during and immediately after infusion, increased sodium, water, and urea excretions from the infused but not from the contralateral kidney were seen. This increase was the same in all groups throughout the infusion; however, in groups infused with EXP 3174, it persisted even after the infusion had been discontinued. Thus, the total amounts of sodium and water excreted during the whole experiment were higher when EXP 3174 was present in the infusion solution than after PD 123319 alone. Moreover, an increase in glomerular filtration rate (creatinine clearance) and renal blood flow (electromagnetic flowmeter) was seen during the infusion period in groups receiving EXP 3174, but not PD 123319. Although it is possible to assume that blockade of AT-1 receptors only is responsible for the changes in excretion rates as PD 123319 is known to interact with these receptors at high concentrations, it is difficult to explain why it does not react with the same receptors responsible for changes in glomerular filtration rate and renal blood flow. An alternative explanation assumes that the excretory changes are due to blockade of both AT-1 and AT-2 receptors; the identity in effect and the absence of additivity of both antagonists could be explained by the existence of a final common pathway leading to the inhibition of the Ang II effect. This pathway could be reached either from AT-1 or AT-2 receptors. PMID- 9419045 TI - Cation conductance regulated by ambient Cl- in cultured rat mesangial cells. AB - Several functions of mesangial cells, such as contraction and release of nitric oxide, are dependent on the ambient Cl- concentration. Herein we describe a direct effect of Cl- on mesangial cell membrane conductance. Rat mesangial cells were isolated, cultured, and were patch-clamped in the whole-cell configuration. The external concentration - ([Cl-]o) - of standard Ringer's solutions was changed isotonically from standard of 160 mM to 35 mM while cell membrane conductance was continuously recorded. Results indicated that control I-V curves of the steady state current had a linear slope conductance of 0.95 nS (n = 10, r = 0.9). In low [Cl-]o, conductance outwardly rectified and increased to 4.1 nS between -120 and 0 mV and to 16.8 nS between 0 and 40 mV (n = 10, r = 0.9; p<0.01). The increase in conductance occurred within a few seconds and was reversible. Our results show that a large mesangial cell membrane conductance for cations is activated upon exposure to low [Cl-]o. PMID- 9419046 TI - Localization of low-KM cAMP phosphodiesterase in rat nephron segments. AB - To evaluate the roles of cAMP degradation on hormonal actions in the kidney, we examined the segmental distribution and activities of cAMP-phosphodiesterase (PDE), a key enzyme for cAMP hydrolysis, in dissected segments of the rat nephron and characterized the isozyme compositions with subtype-specific inhibitors. Summary of the nephron distribution of PDE activities showed that cAMP-PDE activities were detected in all nephron segments and the distal convoluted tubules (DCTs) had the highest activity. Both in proximal convoluted tubules (PCTs) and medullary collecting ducts (MCDs), more than 80% of the total cAMP-PDE activities were inhibited by a nonspecific PDE inhibitor, 3-isobutyl-1 methylxanthine (IBMX). In PCTs, rolipram (type-IV PDE inhibitor) was an equally potent inhibitor of IBMX, while 8-methoxymethyl-IBMX (MM-IBMX, type-I PDE inhibitor) and cilostamide (type-III PDE inhibitor) inhibited cAMP-PDE by 35 and 57%, respectively. In MCDs, inhibition by rolipram (40%) was less than that by MM IBMX (70%) or cilostamide (66%). Rolipram potentiated the parathyroid hormone (PTH)-induced cAMP content in PCTs most effectively. Rolipram and MM-IBMX increased the vasopressin (AVP)-stimulated cAMP content equally in MCDs. Cilostamide had no effect on the cAMP content stimulated by PTH or AVP in PCTs and MCDs. These results indicate that PDE activities were unevenly distributed along the rat nephron and cAMP-PDE was composed of at least three isoforms, namely type I, III and IV in PCTs and MCDs. Among these, type-IV PDE was responsible for hydrolysis of cAMP in PCTs, and type-I and IV PDE were responsible for that in MCDs. PMID- 9419047 TI - Performance of the Amyotrophic Lateral Sclerosis Functional Rating Scale (ALSFRS) in multicenter clinical trials. AB - The Amyotrophic Lateral Sclerosis Functional Rating Scale (ALSFRS) is a 10-item functional inventory which was devised for use in therapeutic trials in ALS. Each item is rated on a 0-4 scale by the patient and/or caregiver, yielding a maximum score of 40 points. The ALSFRS assesses patients' levels of self-sufficiency in areas of feeding, grooming, ambulation and communication. Rotated factor analysis of the ALSFRS found that the rating items group logically and consistently into four categories. The ALSFRS has been validated both cross-sectionally and longitudinally against muscle strength testing, the Schwab and England ADL rating scale, the Clinical Global Impression of Change (CGIC) scale, and independent assessments of patient's functional status. In this report, we use the data provided by the placebo patients who participated in the ALS CNTF Treatment Study (ACTS) to demonstrate the robustness, test-retest reliability and consistency of the ALSFRS as employed in a large, multicenter clinical trial. PMID- 9419048 TI - Amyotrophic lateral sclerosis: mortality risk during the course of the disease and prognostic factors. The Netherlands ALS Consortium. AB - We performed a historical cohort study of 307 untreated patients with probable or definite amyotrophic lateral sclerosis in order to investigate whether the mortality risk changed during the disease course and to identify prognostic factors at diagnosis. Patients were diagnosed in one of the academic hospitals in The Netherlands and followed-up for at least 6 years after diagnosis. The median survival from diagnosis was 1.4 years (95% confidence interval, 1.3-1.6 years) with an estimated 5- and 10-year survival of 20 and 8%, respectively. Mortality was at its maximum in the second year after diagnosis and declined considerably thereafter. Observed mortality approached the expected mortality in patients who survived diagnosis 6 or more years. In univariate and multivariate analyses, young age, limb onset, and a long delay between initial weakness and diagnosis were associated with lower mortality. The better prognosis of limb-onset patients was not observed in females. Patients with initial respiratory muscle weakness, had the worst prognosis with a median survival of only 2 months. The significantly greater mortality of older patients proved not to result from a rise in expected mortality only. In conclusion, the annual mortality risk in ALS does not remain constant throughout the disease and depends on age at diagnosis, site of onset, diagnostic delay, but also on the time since diagnosis. These findings may have consequences for the planning of symptomatic care and the design and analysis of therapeutic trials. PMID- 9419049 TI - A brief quality-of-life measure for ALS clinical trials based on a subset of items from the sickness impact profile. The Syntex-Synergen ALS/CNTF Study Group. AB - We previously demonstrated a significant relationship (P<0.0001) between maximum voluntary isometric contraction (MVC) plus pulmonary function scores (the Tufts Quantitative Neuromuscular Exam Combination Megascore (TQNE CM)), and the Sickness Impact Profile (SIP) in a cohort of 524 ALS patients. Because the 136 item SIP questionnaire can be difficult to administer in this population, we examined SIP subscales and clinically derived item sets in relation to the TQNE CM in an effort to define a briefer measure of quality of life for use in clinical trials. Two 'Mini-SIP' indices performed as well as the overall SIP in reflecting the impact of muscle weakness on ALS patients' quality of life: a combination of two SIP subscales ('SIP-33'), and a 19-item set of questions independently chosen by a panel of ALS specialists ('SIP/ALS-19'). Either index potentially could be useful in ALS clinical trials. The SIP/ALS-19 is currently being used in a National ALS data base, providing an opportunity to evaluate its utility prospectively against other QOL measures in ALS patients. PMID- 9419050 TI - Amyotrophic lateral sclerosis: lessons in trial design from recent trials. AB - The recent history of clinical trials in ALS is marked by a great diversity in trial design. This has limited the comparability of the trial results, and has resulted in intense discussions about which parameters should or should not be evaluated when testing a new drug for ALS. This article discusses, without any claim to completeness, some aspects of trial design in relation to past, ongoing and future ALS trials, including: choice of endpoints, measurement of disease progression, assessment of quality of life, placebo-controlled trials, open-label trials, collaboration with the industry, and methods for investigating mechanisms of action of experimental drugs. PMID- 9419051 TI - Polysomnographic studies in amyotrophic lateral sclerosis. AB - We retrospectively reviewed 17 polysomnograms (PSG) in symptomatic amyotrophic lateral sclerosis (ALS) patients to assess the type and frequency of sleep disordered events and correlated these findings with pulmonary function tests (PFTs), presenting complaints, presence of bulbar dysfunction, and response to bi level positive airway pressure (PAP) treatment. PSG revealed abnormalities in 16 patients. Complaints of orthopnea, daytime sleepiness (but not morning headaches) and a low negative inspiratory force (NIF) correlated with sleep disruption. However, neither the forced vital capacity (FVC) nor the NIF reliably predicted any specific PSG finding. Twelve of 13 patients treated with bi-level PAP responded favorably. Since the response to bi-level PAP is frequently gratifying, PSG should strongly be considered in ALS patients with suspected sleep disturbances. PMID- 9419052 TI - Relative validity of clinical techniques for measuring the body composition of persons with amyotrophic lateral sclerosis. AB - Tracking body composition is necessary to understand how amyotrophic lateral sclerosis (ALS) is affecting a patient's morphology and to provide a basis for appropriate nutritional advice throughout disease progression. Dual X-ray absorptiometry (DEXA) has been shown to reliably detect body composition changes in persons with ALS. However, this procedure is expensive and available primarily for research. The purpose of this study was to determine the relative validity of two common clinical techniques, anthropometry and bioelectrical impedance analysis (BIA), for measuring the body composition of persons with ALS. Twenty three persons with ALS volunteered for the study; seven with primarily bulbar symptoms, five with primarily arm weakness, five with primarily leg weakness, and six with significant weakness in all extremities. On a single day subjects underwent body composition analysis by the three techniques, with DEXA serving as the criterion method. Anthropometry and BIA results were converted to lean and fat mass using eight prediction equations commonly cited in the literature. Anthropometry measures were also converted to estimates of muscle mass using two additional equations. Both BIA and anthropometry tended to overestimate lean mass and underestimate fat mass compared to DEXA. However, the BIA prediction equations had smaller mean differences, larger correlations, and smaller standard errors of estimate than the anthropometry equations. The Lukaski et al. BIA equation (Lukaski, H.C., Bolonchuk, W.W., Hall, C.B., Siders, W.A., 1986. Validation of tetrapolar bioelectrical impedance method to assess human body composition. J. Appl. Physiol. 60, 1327-1332) most closely matched the values derived by DEXA and is probably the best method for measuring the lean and fat mass of persons with ALS, as long as they maintain adequate hydration levels. The Heymsfield et al. equation (Heymsfield, S.B., McManus, C., Smith, J., Stevens, V., Nixon, D.W., 1982. Anthropometric measurement of muscle mass: revised equations for calculating bone-free arm muscle area. Am. J. Clin. Nutr. 36, 680 690) for estimating muscle mass may also be a useful clinical tool for this population. Further longitudinal studies are needed to determine whether the equations that correlated best with DEXA at a single point in time are also sensitive enough to detect changes in body composition over a period of time. PMID- 9419053 TI - Fasciculations: what do we know of their significance? AB - Fasciculations are observed in patients with neurogenic disorders and in healthy individuals. Depending on the associated clinical symptoms and signs, they may signify the presence of a variety of disorders of the lower motor neuron. Divergent and occasionally conflicting opinions prevail regarding the aetiology, pathogenesis, clinical significance, neurophysiological characteristics and the physiological site of origin of fasciculations. In this review we examine the published literature and attempt to clarify these issues. PMID- 9419054 TI - MR spectroscopy in amyotrophic lateral sclerosis/motor neuron disease. AB - Proton magnetic resonance spectroscopy (1H-MRS) and proton magnetic resonance spectroscopic imaging (1H-MRSI) have been used to identify neuronal dysfunction and/or loss in vivo in patients with various neurological diseases, including amyotrophic lateral sclerosis/motor neuron disease (ALS/MND). Both long and short echo time (TE) proton spectroscopy reveal the brain metabolites choline (Cho), creatine/phosphocreatine (Cr), and N-acetyl (NA) groups. Because NA groups are localized to mature neurons and Cr is homogeneously distributed throughout the brain, the NA/Cr ratio is considered an index of neuronal integrity. Long TE proton spectroscopic studies have revealed significantly decreased NA/Cr values in the sensorimotor cortex and brainstem of patients with ALS, consistent with neuronal dysfunction and/or loss. The amount of NA/Cr decrease appears to be directly proportional to the degree of clinical upper motor neuron deficit. Short TE 1H-MRS and 1H-MRSI also reveal other metabolites such as glutamate (Glu) and glutamine (Gln), which have been implicated in the ALS/MND disease process. Preliminary results of short TE 1H-MRSI of the medulla in patients with ALS/MND have revealed significantly decreased NA/Cr values and abnormally elevated Glu+Gln/Cr ratios, compared to control individuals. The latter values were higher in patients with more rapid disease. Although it is unclear whether the elevation of Glu+Gln/Cr precedes or follows the neuronal (and axonal) degeneration in the medulla of these patients, its occurrence provides in vivo evidence of abnormal glutamate metabolism in the CNS parenchyma of patients with ALS/MND. PMID- 9419056 TI - Toxicity of cysteine and cysteine sulphinic acid to human neuronal cell-lines. PMID- 9419055 TI - In vivo and in vitro studies of glycine- and glutamate-evoked acetylcholinesterase release from spinal motor neurones: implications for amyotrophic lateral sclerosis/motor neurone disease pathogenesis. AB - To investigate the spinal cellular structures and molecular mechanisms involved in acetylcholinesterase (AChE) release evoked by both glycine (GLY) and glutamate (GLU)--responses that might play a role in chronic neurotoxicity--we analysed AChE histochemistry and histology upon systemic administration of aspartate (ASP), and conducted in vitro experiments in synaptosomes and slices prepared from mouse spinal ventral horns. Upon superfusion and incubation exposure of these preparations to GLY- and GLU-receptor agonists, we assayed both tissue content and release of AChE, butyrylcholinesterase and lactic dehydrogenase. Histochemical reduction of motor neurone (MN) AChE, calcium dependency, decreases in intracellular AChE and the ratio amongst molecular forms released, suggest that both synaptosomal GLY-evoked AChE release (GLY-EAR) and GLU-receptor elicited AChE release (GEAR) have release sites located at MN presynaptic terminals. These responses exhibited remarkable postnatal regulation. GEAR seems to be mediated through alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid/kainate receptors after the fourth postnatal week and through both NMDA and non-NMDA receptors at earlier stages. Sustained rises of extracellular AChE might link acute excitotoxic injury with several long-lasting pathways leading to chronic neurotoxicity, since AChE molecular properties include: (1) the ability to block cholinergic mechanisms that protect MN against overactivity; (2) activation of ATP-dependent potassium channels; (3) promotion of neurite and axon outgrowth; and possibly (4) stimulation of brain macrophage migration and activation. PMID- 9419057 TI - The use of transgenic mouse models of amyotrophic lateral sclerosis in preclinical drug studies. AB - The discovery of mutations in the human SOD1 gene encoding Cu,Zn superoxide dismutase (Cu,Zn SOD) in patients with familial amyotrophic lateral sclerosis (ALS) has made possible the development of etiological models of the disease. Expression of mutant SOD1 genes in transgenic mice causes a progressive paralytic disease whose general features resemble ALS in humans. We have used the transgenic model to explore etiological mechanisms and to screen potential therapeutics. Our results and those of others show that familial ALS mutations cause a gain-of-function in Cu,Zn SOD that enhances the generation of damaging oxygen radicals. This may render motor neurons sensitive to the excitotoxic effects of ambient glutamate, as a putative glutamatergic inhibitor such as riluzole has therapeutic efficacy both in the transgenic model and in human ALS. This finding highlights the utility of the SOD1-G93A transgenic mouse model for preclinical drug studies in ALS. PMID- 9419058 TI - Hereditary canine spinal muscular atrophy: genetics, neurophysiology, and pathology. PMID- 9419059 TI - Assessment of depression in patients with motor neuron disease and other neurologically disabling illness. AB - Motor neuron disease (MND) is a severely disabling, relentlessly progressive neurological condition with a marked reduction in life expectancy which might be expected to be associated with significant depression and psychological dysfunction following diagnosis. There is very little data on the incidence of depression in MND and most of the published evidence would suggest that depression is rare in patients with MND, especially when compared to other neurologically disabling illnesses. We have studied 40 patients with MND and compared them to a group of 92 patients with multiple sclerosis (MS) attending a neurology clinic. Depression was assessed using the Beck Depression Index (BDI) and the Hospital Anxiety and Depression Scale (HAD). There was no difference in incidence or severity of depression in these two patient groups. For the whole study group there was no difference in depression scores when compared by age, gender or marital status. Depression scores showed a weak association with increasing physical disability measured by the SF36 Physical Function scale for the MS group but there was no association between depression levels and SF36 physical function for the MND group. The MND group did, however, show a significant association between depression scores and pain measured by the SF36 scale. Anxiety levels (HAD scale) were shown to be significantly higher in females and married (versus single, widowed or divorced) subjects for the group as a whole. We conclude that depression is at least as common in MND patients as other neurologically disabled patients with MS and may often be associated with significant pain. Physicians and others involved in the care of patients with MND should be aware that depression and pain may be significant problems irrespective of the level of physical disability. PMID- 9419060 TI - Factors influencing a patient's decision regarding riluzole: an early experience. AB - Riluzole is the first drug to be approved in the United States for the treatment of amyotrophic lateral sclerosis (ALS). During the first 8 months of the drug's availability by prescription, its use was discussed with 46 patients with probable or definite ALS as defined by the E1 Escorial criteria. Seventeen of the patients (37%) chose to take riluzole while 29 (63%) refused. Increased duration of symptoms, increased time since diagnosis, and participation in either the insulin-like growth factor-1 (IGF-1) and brain-derived neurotrophic factor (BDNF) trials were all associated with decreased likelihood of starting the drug. The most common reason given for not wanting to take the medication was insufficient benefit. PMID- 9419061 TI - Home care of patients with amyotrophic lateral sclerosis (ALS). AB - Optimal home care maximizes function and quality of life for patients with ALS. We designed a survey to study home care in the ALS population. Ninety-eight patients with ALS completed our survey. Of these, 24 receive non-hospice home care, nine hospice home care, and seven both hospice and non-hospice home care. Fifty-eight patients receive no outside help. Patients receiving hospice are older than those receiving non-hospice home care (68.9 vs. 57.7 years, P<0.05). Patients with home care assistance have a mean ALS Functional Rating Scale (ALS FRS) score of 13, and those without home care assistance have a mean score of 26 (P<0.0001). Patients receiving non-hospice home care assistance have a median of 16 h/week of care, while those with hospice receive 5.5 h/week (P=0.05). Patients on Medicaid receive more hours of home care than those with any other insurance (median 61 vs. 3.4 h/week with Medicare and 5 h/week with commercial insurance, P=0.008). Primary caregivers spend a median of 11 h/day caring for patients despite having home care assistance. Forty-two and 48% of primary caregivers feel physically and psychologically unwell, respectively. Home care received by patients with ALS often is inadequate and too late to relieve the burden placed on family caregivers. PMID- 9419062 TI - The amyotrophic lateral sclerosis (ALS) support network of Kentucky: an informational support group using interactive video. AB - Support groups are beneficial to ALS patients and their families by providing a forum for discussion and emotional support. The low prevalence of ALS and the patients limited mobility make it difficult to sustain support groups in rural environments. We describe the organization of an ALS support network in the Commonwealth of Kentucky using real-time, interactive video. PMID- 9419063 TI - End of life decisions in amyotrophic lateral sclerosis: a cross-cultural perspective. AB - In an era where life-sustaining technology offers physicians unprecedented powers to prolong survival in terminal illness, the question of how end of life decisions are made has become a major subject of study and debate. Amyotrophic lateral sclerosis (ALS) is a disease in which physical ability declines while mental capacity most often remains intact. Since most patients with ALS die of respiratory failure, a distinguishing feature of this disease is whether a patient is offered and accepts a chance to go on long-term mechanical ventilation. This unusual feature makes ALS a compelling model for studying end of life decisions in different countries. This paper reviews the literature and presents preliminary data on how end of life decisions in ALS are made in the US, Great Britain, and Japan. We address this issue by examining how cultural differences in truth-telling and informed consent, societal differences in attitudes toward the use of artificial life support, and legal differences in the role and status of advance directives in each country influenced decisions in the following three groups of patients: (1) the mentally competent; (2) mentally incompetent patients who previously completed advance directives when competent; and (3) mentally incompetent patients who have not provided advance guidance about their wishes. PMID- 9419064 TI - Ventilatory support: Japanese experience. AB - Paternalism is a predominant pattern in the social and behavioural climate of medical practice in Japan. Recognition of the sanctity of life is the first principle guiding medical ethics. This principle is an important factor in considering the management of patients with ALS in Japan. Following discussion of these principles in the application of ventilatory support to patients with ALS among the Ministry of Health and Welfare of Japan (MHWJ), the Japan ALS Association (JALSA) and ALS patients and families in the Tokyo Metropolitan Neurological Hospital (TMNH), I have concluded that the development of ventilatory failure in ALS in the Japanese context should not be regarded as the endpoint of the disease but merely one of its impairments. Ventilatory support, considered in this context, can extend the life of an ALS patient. PMID- 9419065 TI - Analysis of the step response of the saccadic feedback: computational models. AB - We present results of theoretical analysis and computational simulations of two models of the saccadic burst generator: the Scudder model and the Jurgens model. We used the experimental paradigm of prolonged stimulation in monkey superior colliculus (SC) to compare the performance of the two models. We excluded the Scudder model since it was not capable of reproducing the experimentally observed staircase movements. We modified the Jurgens model by replacing the originally proposed feedback integrator with an active reset mechanism by a leaky integrator. With this modification we have shown that the staircase movement elicited by prolonged stimulation in the SC can be modeled as a damped oscillatory step response of this model. Furthermore, to replicate the changes in the kinetic profiles of the staircase movements with increased stimulation we modified the functionality of the model. Our results suggest that prolonged stimulation of the SC dynamically changes the gains and time constant of the saccadic feedback. PMID- 9419066 TI - Somatotopic organization along the central sulcus, for pain localization in humans, as revealed by positron emission tomography. AB - Regional cerebral blood flow was measured with positron emission tomography (PET) in six healthy volunteers at rest and during experimentally induced, sustained cutaneous pain on the dorsum of the right hand or on the dorsum of the right foot. Pain was inflicted by intracutaneous injection of capsaicin, providing a mainly C-fibre nociceptive stimulus. Statistical analysis showed significant activations along the central sulcus (SI) area when comparing pain in the hand to pain in the foot. Separate comparison of both pain states to a baseline revealed different locations along the central sulcus for hand pain and foot pain. The encountered differences are consistent with what is previously known about the somatotopics of non-painful stimuli. When comparing painful stimuli to baseline, the contralateral anterior cingulate gyrus, the ipsilateral anterior insular cortex and the ipsilateral prefrontal cortex were implicated. The results are consistent with an involvement of SI in the spatial discrimination of acute cutaneous pain. PMID- 9419067 TI - Effects of 17beta-estradiol on glucose transporter 1 expression and endothelial cell survival following focal ischemia in the rats. AB - Estrogen replacement therapy in postmenopausal women is associated with a decreased mortality and morbidity from stroke. The present study was undertaken to investigate the effects of estrogen on endothelial cell glucose transporter 1 (GLUT 1) and on the cell viability during focal ischemia in a rat model. Female rats were ovariectomized (OVX) and 2 weeks later 17beta-estradiol (E2) was injected subcutaneously at a dose of 100 microg/kg 2 h before unilateral middle cerebral artery (MCA) occlusion. Ischemic lesion size was quantified using 2,3,5 triphenyl tetrazolium chloride (TTC) staining and GLUT 1 protein was analyzed by Western blotting. E2 treatment decreased ischemic lesion size in slices taken at 9 and 11 mm posterior from the olfactory bulb by 46.3% and 44.1%, respectively (P < 0.05). GLUT 1 protein decreased in both OVX and E2 groups by 24.6% and 22.7% respectively (P < 0.05) compared with the non-lesioned side in the core ischemic region, including the basal ganglia. GLUT 1 protein was increased in the E2 treated group compared with the control group (23.3%, P < 0.05) in the penumbral ischemic region of the cortex. Primary rat brain capillary endothelial cell (BCEC) cultures were established as an in vitro model for ischemic effects on endothelial cells. Estrogen reduced BCEC loss by 35.9%, 28.4% and 23.5% (P < 0.05) when glucose in the culture medium was reduced to 50%, 20% and 10%, respectively; and by 28.4% and 18.4% (P < 0.05) following 1 or 4 h of anoxia, respectively. This study demonstrates that estrogen treatment increases GLUT 1 transporters and protects BCEC loss which may in turn reduce focal ischemic brain damage. PMID- 9419068 TI - Dopaminergic modulation of neuronal activity in the monkey putamen through D1 and D2 receptors during a delayed Go/Nogo task. AB - Using multibarreled glass micropipettes, we recorded single-unit activity in the putamen, and iontopho retically applied D1 and D2 dopamine receptor agonists (SKF38393, quinpirole) and antagonists (SCH23390, sulpiride) while two monkeys were performing a delayed Go/Nogo task. The putaminal neurons exhibited changes in activity during various task periods (hold, cue, delay, response, and reward periods) in both Go and Nogo trials. Of 296 task-related putaminal neurons, 87 showed activity changes in Go trials only (Go type), 74 in Nogo trials only (Nogo type), 99 in both trials during the same task periods (Both type), and 36 in both trials but during different task periods (Different type). These 296 neurons were examined as regards the effects of both D1 and D2 agonists and/or antagonists, and 234 neurons responded to either D1 - or D2-related substances or both. Among them 41% of neurons responded to the D1 substances only (D1 group), 36% responded to the D2 substances only (D2 group), and 23% responded to both D1 and D2 substances (D1D2 group). During the iontophoretic application of the D1 and D2 substances, most of the responding neurons changed their task-related activity but not their baseline firing rates. The D1 agonist increased the activity in 19 neurons and decreased it in 105 neurons. On the other hand, the D2 agonist increased the activity in 54 neurons and decreased it in 50 neurons. The D1 and D2 substances modulated the activity in both Go and Nogo trials. Each of the three D1/D2 groups (D1, D2, and D1D2 groups) contained all four Go/Nogo types (Go, Nogo, Both, and Different types) of neurons. Percentages of each Go/Nogo type of neuron were comparable among the three D1/D2 groups. The D1 and D2 substances modulated the activity related to various task periods. Each of the three D1/D2 groups included neurons activated during the cue, delay, response, or reward period in Go and Nogo trials. Distributions of the neurons related to each task period were similar among the D1/D2 groups. These results suggest that dopamine can modulate the activity of single putaminal neurons through both D1 and D2 receptors and that the dopaminergic modulation through the two receptors in the putamen affects similar types of signals in behavioral control. PMID- 9419069 TI - Corticocortical connections between visual areas 17 and 18a of the rat studied in vitro: spatial and temporal organisation of functional synaptic responses. AB - Much is known about the anatomy of corticocortical connections, yet little is known concerning their physiology. In order to have access to the synaptic and temporal aspects of the activity elicited through corticocortical connections, we developed an in vitro approach on slices of rat visual cortex. We used extracellular recordings of field potentials combined with electrical stimulation to localise regions of areas 17 and 18a that are connected. We found that corticocortical connections between areas 17 and 18a can be preserved in 500 microm thick slices, with a focus of activity separated from the stimulating electrode by 1.5 mm to more than 3 mm. The potentials elicited in one area after stimulation of its neighbour displayed fast events, corresponding to action potentials, and slow events, corresponding to synaptic potentials. Intracellular recordings showed that the earliest synaptic responses consisted of monosynaptic excitatory potentials. Measurement of response latency showed that axons involved in both feedforward and feedback corticocortical connections are slowly conducting (0.3-0.8 m/s). Conduction velocity for antidromically activated cells was not significantly different for the two sets of connections. In an attempt to establish the spatial organisation of functional synaptic inputs, field potential recordings were performed in the different cortical layers and used to establish current source density (CSD) graphs along the depth axis. The CSD maps obtained were found to be somewhat variable from one case to another. It is suggested that this variability results from the use of electrical stimulation, which activates axons that are both afferent and efferent to a given cortical area. The field potentials are therefore likely to contain responses that correspond to the activity mediated by the intrinsic collaterals mixed in variable amount with responses produced by corticocortical synapses. With this restriction in mind, it is suggested that, after stimulation of the supragranular layers, the functional synaptic inputs of feedforward connections are concentrated in layer 4 and the bottom of layer 3, while those of feedback axons involve mainly the upper part of the supragranular layers. The intrinsic collaterals of the neurones participating in corticocortical connections seem also to provide the bulk of their inputs to the upper part of the supragranular layers. The laminar pattern of activity obtained after infragranular layer stimulation was comparable to that obtained after supragranular layer stimulation, except for the addition of a supplementary region of activated synapses in the infragranular layers. PMID- 9419070 TI - Response properties of pigeon otolith afferents to linear acceleration. AB - In the present study, the sensitivity to sinusoidal linear accelerations in the plane of the utricular macula was tested in afferents. The head orientation relative to the translation axis was varied in order to determine the head position that elicited the maximal and minimal responses for each afferent. The response gain and phase values obtained to 0.5-Hz and 2-Hz linear acceleration stimuli were then plotted as a function of head orientation and a modified cosine function was fit to the data. From the best-fit cosine function, the predicted head orientations that would produce the maximal and minimal response gains were estimated. The estimated maximum response gains to linear acceleration in the utricular plane for the afferents varied between 75 and 1420 spikes s-1 g-1. The mean maximal gains for all afferents to 0.5-Hz and 2-Hz sinusoidal linear acceleration stimuli were 282 and 367 spikes s-1 g-1, respectively. The minimal response gains were essentially zero for most units. The response phases always led linear acceleration and remained constant for each afferent, regardless of head orientation. These response characteristics indicate that otolith afferents are cosine tuned and behave as one-dimensional linear accelerometers. The directions of maximal sensitivity to linear acceleration for the afferents varied throughout the plane of the utricle; however, most vectors were directed out of the opposite ear near the interaural axis. The response dynamics of the afferents were tested using stimulus frequencies ranging between 0.25 Hz and 10 Hz (0.1 g peak acceleration). Across stimulus frequencies, most afferents had increasing gains and constant phase values. These dynamic properties for individual afferents were fit with a simple transfer function that included three parameters: a mechanical time constant, a gain constant, and a fractional order distributed adaptation operator. PMID- 9419071 TI - Torsion during saccades between tertiary positions. AB - We systematically studied the effect of saccade direction and saccade starting position on the velocity profile of the saccade. Saccades were made between targets placed at optical infinity by dichoptic presentation. This arrangement was chosen to evoke conjugate eye movements. Eye movements were recorded binocularly, including torsion. Horizontal and vertical movements of the eyes are strongly correlated (r > or = 0.95) during the saccade, torsional movements are much less so (r approximately 0.67). Listing's law would predict that the three dimensional versional velocity of the eye would be located in a plane that is tilted out of Listing's plane by an amount that depends on the saccade's starting position (half angle rule). Taking together all saccades that started from the same initial position a plane could be fitted through the velocity vectors. However, this plane was tilted less relative to Listing's plane than predicted by the half angle rule. The deviation was especially large for the yaw component of the tilt (56% of predicted). For the pitch component the prediction was better (81% of predicted). In addition, we find that the torsional velocity during the fast "intrasaccadic" part of the motion can be unequal in the two eyes. The implications for three-dimensional models of saccadic control are discussed. PMID- 9419072 TI - The functional effectiveness of neck muscle reflexes for head-righting in response to sudden fall. AB - Reflex head-righting in normal and labyrinthine-defective (LD) subjects was compared to identify the relative functional effectiveness of vestibular-collic and cervico-collic myotactic reflexes. To restrict stimuli largely to the head and neck, subjects lay supine, supported up to the shoulders on a horizontal bed with their head supported in a sling over the edge. The head fell freely as the sling was released with an electromagnetic catch. Head drops were delivered with the subjects instructed to relax and accept the fall passively or to actively right the head as fast as possible. With both instructions, righting responses in normal subjects commenced with electromyographic (EMG) bursts in the sternocleidomastoid (SCM) at 24.5 ms latency, which was reflected in a deceleration of the downwards head velocity. The latency of the earliest EMG responses in LD subjects was 67.4 ms, accompanied by similar deceleration. It is assumed that the earliest response in normal subjects is vestibular, whereas in LDs the SCM stretch reflex is the earliest response. These reflexes are followed at circa 100 ms by more intense EMG activity due to voluntary movement, but braking of head fall is evident before voluntary activity takes effect. Righting was more effective in normal subjects than in LDs, and when "active" normal subjects made more vigorous righting responses than when "passive"; whereas active righting in LDs was no better than passive. The results demonstrate that reflex responses contribute significantly to head-righting. The vestibular contribution gives an advantage over stretch reflexes alone and also assists in voluntary enhancement of reflex responses. PMID- 9419073 TI - Time to contact and the control of manual prehension. AB - In the present study, a kinematic analysis was made of unconstrained, natural prehension movements directed toward an object approaching the observer on a conveyor belt at one of three constant velocities, from one of three different directions (head-on or along the fronto-parallel plane coming either from the subject's left or right). Subjects were required to grasp the object when it reached a target located 20 cm directly in front of the hand's start position. The kinematic analysis revealed that both the transport and grasp components of the movement changed in response to the experimental manipulations, but did so in a manner that guaranteed that, for objects approaching from a given direction, hand closure would begin at a constant time prior to object contact (regardless of the object's approach speed). The kinematic analysis also revealed, however, that the onset of hand closure began earlier with objects approaching from the right than from other directions -- an effect which would not be predicted if time to contact was the key variable controlling the onset of hand closure. These results, then, lend only partial support to the theory that temporal coordination between the transport and grasp components of prehension is ensured through their common dependence on time to contact information. PMID- 9419074 TI - Linear and nonlinear properties of simple cells of the striate cortex of the cat: two types of nonlinearity. AB - In a proportion of simple cells of the striate cortex, the weighting functions of the receptive fields (RFs) had more periods than could be established by mapping using responses to light bars and dark bars. In these multiperiodical cells, side subfields do not respond to single bars, as they have lower weights than central zones and the excitation is under the threshold of impulse response if a single bar is applied. This fact has been established by different methods: conditioning and testing stimuli, grating patches, and inverse Fourier transform of the amplitude-phase characteristic, combining them in one cell. We assume that this type of nonlinarity can be used in analyzing the image, as it acts as a spatial frequency filter of the area overlapped by the RF. The responses to complex gratings composed by two sinusoidal gratings of different frequency, contrast, and phase shift were compared with the sum of the responses to the gratings when they were presented separately. The results show that the principle of superposition holds a reasonable approximation even if the response is evoked from the side subzones. Some simple cells have nonlinear properties beyond the classic zone of RF (2nd type of nonlinearity). Linear cells have a tendency to be localized in layer 4 of striate cortex, cells with a nonlinear surround in layers 2, 3, 5, and 6. The significance of both types of nonlinearities in simple cells is discussed. PMID- 9419075 TI - Parietal cortex and movement. I. Movement selection and reaching. AB - Recording studies in the parietal cortex have demonstrated single-unit activity in relation to sensory stimulation and during movement. We have performed three experiments to assess the effect of selective parietal lesions on sensory motor transformations. Animals were trained on two reaching tasks: reaching in the light to visual targets and reaching in the dark to targets defined by arm position. The third task assessed non-standard, non-spatial stimulus response mapping; in the conditional motor task animals were trained to either pull or turn a joystick on presentation of either a red or a blue square. We made two different lesions in the parietal cortex in two groups of monkeys. Three animals received bilateral lesions of areas 5, 7b and MIP, which have direct connections with the premotor and motor cortices. The three other animals subsequently received bilateral lesions in areas 7a, 7ab and LIP. Both groups were still able to select between movements arbitrarily associated with non-spatial cues in the conditional motor task. Removal of areas 7a, 7ab and LIP caused marked inaccuracy in reaching in the light to visual targets but had no effect on reaching in the dark. Removal of areas 5, 7b and MIP caused misreaching in the dark but had little effect on reaching in the light. The results suggest that the two divisions of the parietal cortex organize limb movements in distinct spatial coordinate systems. Area 7a/7ab/LIP is essential for spatial coordination of visual motor transformations. Area 5/7b/MIP is essential for the spatial coordination of arm movements in relation to proprioceptive and efference copy information. Neither part of the parietal lobe appears to be important for the non-standard, non-spatial transformations of response selection. PMID- 9419076 TI - Parietal cortex and movement. II. Spatial representation. AB - Lesions in the two divisions of parietal cortex, 5/7b/MIP and 7a/LIP, produce dissociable reaching deficits. Monkeys with 5/7b/MIP removals were tested on reaching in the dark under two different conditions. All the reaches made on any day were from the same starting position to the same target position in the control condition. In the "transfer" condition, all the reaches were made to the same target position but consecutive reaches were made from different starting positions. The target could be represented as a constant pattern of joint and muscle positions in the control condition. The transfer condition required a representation of the starting position of the hand and/or a representation of the target in terms of its position in space. Removal of areas 5, 7b and MIP produced only a very mild impairment in the control condition and a severe impairment in the transfer condition. This suggests that 5/7b/MIP does not represent the limb in simple sensory or motor coordinates but in terms of its spatial position. PMID- 9419077 TI - Alterations in mystacial pad innervation in the aged rat. AB - It is well established that sensory perception becomes impaired with advancing age and that, in parallel, dystrophy and degeneration of axons occur in sensory pathways. In this study, the impact of aging was examined in the mystacial pad, which receives a large variety of sensory nerve endings organized in a highly predictable pattern. Mystacial pad specimens from aged (30 months old) and young adult (2-3 months old) female Sprague-Dawley rats were processed, in parallel, for immunohistochemical analyses with antibodies against human neuronal cytoplasmic protein (protein gene product 9.5), transmitter enzymes, and several neuropeptides. Several changes in cutaneous innervation including both degenerative and regenerative processes were evident in the aged rat: (1) the Merkel endings and lanceolate endings that emanate from large-caliber afferents in the whisker follicles were reduced and showed signs of degeneration. Furthermore, a reduction of piloneural complexes at the intervibrissal hairs were evident, but only in aged rats that showed more severe behavioral sensorimotor disturbances. In contrast, Ruffini endings as well as mechanoreceptors emanating from medium-caliber axons, i.e., transverse lanceolate and reticular endings, appeared normal. (2) A reduction was evident among two sets of unmyelinated epidermal endings; however, the epidermal innervation affiliated with the intervibrissal hairs appeared normal in the aged rat. (3) A loss of sympathetic neuropeptide tyrosine (NPY) or tyrosine hydroxylase-immunoreactive (IR) and somatosensory Calcitonin gene-related peptide (CGRP)-IR perivascular axons was paralleled by an increase in presumed parasympathetic NPY/CGRP-IR axons. (4) Two "novel" networks of fine-caliber axons were observed in the outer and inner root sheaths of the whisker follicles in the aged rat. (5) NPY was present in a population of small-caliber, somatosensory CGRP-IR axons in the aged rat. This may represent a de novo synthesis, since, normally, NPY-like immunoreactivity is not observed in this set of axons. Our results suggest that the sensory impairments occurring with advancing age are part of a peripheral process instigated by changes in nerve-target interactions and/or incapacitation of the neuronal machinery to sustain the axonal integrity. PMID- 9419079 TI - The development toward stereotypic arm kinematics during reaching in the first 3 years of life. AB - We recorded reaching movements from nine infants longitudinally from the onset of reaching (5th postnatal month) up to the age of 3 years. Here we analyze hand and proximal joint trajectories and examine the emerging temporal coordination between arm segments. The present investigation seeks (a) to determine when infants acquire consistent, adult-like patterns of multijoint coordination within that 3-year period, and (b) to relate their hand trajectory formation to underlying patterns of proximal joint motion (shoulder, elbow). Our results show: First, most kinematic parameters do not assume adult-like levels before the age of 2 years. At this time, 75% of the trials reveal a single peaked velocity profile of the hand. Between the 2nd and 3rd year of life, "improvements" of hand or joint-related movement units are only marginal. Second, infant motor systems strive to obtain velocity patterns with as few force reversals as possible (uni- or bimodal) at all three limb segments. Third, the formation of a consistent interjoint synergy between shoulder and elbow motion is not achieved within the 1st year of life. Stable patterns of temporal coordination across arm segments begin to emerge at 12-15 months of age and continue to develop up to the 3rd year. In summary, the development toward adult forms of multijoint coordination in goal-directed reaching requires more time than previously assumed. Although infants reliably grasp for objects within their workspace 3-4 months after the onset of reaching, stereotypic kinematic motor patterns are not expressed before the 2nd year of life. PMID- 9419080 TI - Thriving in academic medicine. PMID- 9419078 TI - Gravitational inputs modulate visuospatial neglect. AB - Right brain-damaged patients with left visuospatial neglect were required to bisect a line placed in front of them in two different body positions (upright and supine) and two different light conditions (light and dark). The neglect patients, unlike right brain-damaged patients without neglect, strongly reduced their rightward directional error in the supine compared with the upright position. No systematic changes were produced by the light-dark manipulation. The present result cannot be explained with an attentional interpretation of hemispatial neglect. We suggest that the present data provide further evidence that hemineglect is the consequence of a mismatch between different afferent information integrated into an egocentric space representation. According to this model, the presence of a lateralized brain lesion produces asymmetries in some intermediate spatial representations (eye-head, head-trunk, body-environment) but not in the retinotopic one. Any experimental manipulation that reduces the asymmetry of the intermediate representation such as the reduction of gravitational inputs may improve the dynamic integration of the egocentric coordinates. PMID- 9419081 TI - Clinical observations on coexistence of sudden hearing loss and vestibular schwannoma. AB - It has long been recognized that sudden hearing loss (SHL) may be a harbinger of vestibular schwannoma (VS). Among 192 VS patients who underwent operation in the Gruppo Otologico, Piacenza, Italy, from April 1987 to October 1995, the charts of 14 (7.3%) cases with a history of SHL were examined. SHL was the first symptom in 8 (4.2%) patients. Eight (57.1%) of 14 VS cases with SHL anamnesis had reported recovery of their previous hearing either totally or partially before establishment of tumor diagnosis. Five (35.7%) cases had recurrent bouts of SHL. SHL was observed less frequently in cases with large tumors (>3 cm). However, the frequency of SHL in patients with small tumors did not differ from that of medium sized tumors. Awareness about coexistence of SHL and VS, as well as concomitant use of auditory brain stem response and magnetic resonance imaging, is crucial to rule out the diagnosis of VS in a patient with SHL. PMID- 9419082 TI - Tympanoplasty in young patients: the role of adenoidectomy. AB - A retrospective study of 60 pediatric patients with dry tympanic membrane perforation undergoing type I tympanoplasty during a 15-year period was carried out. Seventy-seven percent of patients were followed up for 5 years. The overall success rate was 90%. All failures occurred in patients who previously had undergone adenoidectomy or adenotonsillectomy. However, sex was found to be the only statistically significant prognostic factor of tympanoplasty success: female patients had higher success rates than male patients. Neither patient age, prior ventilation tube placement, size of perforation, status of the contralateral ear, surgical technique (underlay or overlay), nor competence of the surgeon (resident or senior) affected the success rate. The possible reasons for these findings will be discussed. PMID- 9419083 TI - Removal of jugular foramen tumors: the fallopian bridge technique. AB - Despite recent advances in neuroradiographic and electrophysiologic assessment, the surgical extirpation of lesions of the bony skull base remains challenging. Moreover, as surgeons have gained experience in removing tumors from the irregular osteologic confines of the skull base, attention has been directed toward preservation of vital neural and vascular structures traversing the operative field. This report describes the creation of a fallopian bridge with preservation of the facial nerve in removing tumors that arise within or juxtaposed to the jugular fossa. Thirty-five patients are reported herein with analysis of pathology, surgical approach, and outcome. An algorithm for use of the fallopian bridge, as opposed to facial nerve mobilization and rerouting, is presented with particular emphasis on limitation of this selective procedure. PMID- 9419084 TI - Mastoidectomy in noncholesteatomatous chronic suppurative otitis media: is it necessary? AB - Chronic suppurative otitis media (CSOM) without cholesteatoma, the surgical treatment of which is still controversial, is a common diagnosis in otologic practice. A retrospective analysis of 323 patients who underwent surgery for noncholesteatomatous chronic otitis media in the Gruppo Otologica, Piacenza, Italy, between April 1983 and December 1993 is presented. Cases were separated into three groups according to different surgical treatment modalities and conditions of the ears at the time of operation. Group I (n = 53) consisted of cases of CSOM treated by tympanoplasty without mastoidectomy (TLWOM). Group II (n = 28) included cases of CSOM treated by tympanoplasty with mastoidectomy (TLWM). Intact canal wall technique was used in these cases. The ears in both these groups were discharging severely at the time of surgery. Group III (n = 242) included patients whose ears were dry at the time of surgery but who had had previous recurrent episodes of suppuration and who were treated by TLWOM. At the last follow-up, graft success rates for groups I, II, and III were 90.5%, 85.7%, and 89.2%, respectively, and mean residual gaps were 17.2 dB, 20.1 dB, and 19.4 dB, respectively. There was no statistically significant difference between the three groups either on graft success rates (p > 0.05) or on final functional hearing outcome (p > 0.05). TLWM is the preferable treatment modality for most surgeons in noncholesteatomatous CSOM. Nevertheless, in our experience TLWOM yields comparable results for this group of patients. In addition, we could not find any significant difference in results of graft success and final functional hearing rates between dry and discharging ears (p > 0.05). PMID- 9419086 TI - Are acoustic neuromas encapsulated tumors? AB - In articles and chapters on the subject of acoustic neuroma, it is almost invariably stated that they are well-encapsulated tumors. During surgical procedures, blunt mechanical dissection defines a natural subsurface cleavage plane that leaves intact a several millimeter thick rind of tumor surface. Occasionally, as a concession to neural integrity, less than complete resection is elected, leaving behind this "capsular" remnant. To clarify the nature of the surface of acoustic neuromas and to test whether this long held description is indeed correct, a microscopic analysis of 10 surgical specimens was performed. A wedge was harvested from the free surface of the tumor in the mid cerebellopontine angle that included a large, undisturbed section of the tumor surface. Histologic analysis showed that for most of the tumor surface only an extremely thin (3 to 5 microm) layer of connective tissue envelops the tumor. Neoplastic Schwann cells, which extend essentially to the margin of the tumor, were found to be somewhat flattened and compressed in the vicinity of the surface. Although acoustic neuromas are surrounded by a continuous layer of connective tissue, it is so exceptionally thin (on average less than the diameter of a red blood cell) that its edge cannot be visualized intraoperatively by a surgeon. Because the pathologic definition of a capsule is a thick, enveloping layer of connective tissue that is both micro- and macroscopically evident, it must be concluded that acoustic neuromas are nonencapsulated, at least in the conventional sense of the term. The surface peel observed intraoperatively is surgically produced during tumor debulking by cleaving of the looser central component from the more compressed portion of neoplastic cells that lies immediately beneath the free margin of the lesion. PMID- 9419085 TI - Mechanisms of auditory impairment during acoustic neuroma surgery. AB - Hearing loss during removal of acoustic neuroma (AN) may be due to labyrinthine and/or neural and/or vascular damage. Surgical maneuvers relating to perioperative and postoperative hearing may give rise to mechanisms of auditory impairment. Recording action potentials from the intracranial portion of the cochlear nerve (CN) has proven particularly useful for identifying the mechanisms of iatrogenic auditory injury. In this paper intraoperative and postoperative auditory impairments are investigated in relation to surgical steps in a group of 47 subjects with AN (size ranging from 5 to 25 mm) undergoing removal by a retrosigmoid-transmeatal approach. Drilling of the internal auditory canal (IAC), removal of the AN from the IAC fundus, coagulation close to the CN, lateral to medial tumor traction, separation of the CN from the facial nerve, and stretching of the CN have proven to be the most critical surgical steps in hearing preservation. On the other hand, maneuvers such as intracapsular tumor removal, vestibular neurectomy, suction close to the AN, and closure of the IAC defect did not correlate with changes in auditory potentials. Predisposing factors to postoperative hearing deterioration were IAC enlargement greater than 3 mm, IAC tumor size greater than 7 mm, extracanalar tumor size greater than 20 mm, labyrinth medial to the IAC fundus, severe involvement of the CN in the IAC, preoperative abnormal auditory brainstem responses, and normal vestibular reflectivity. Age and preoperative hearing did not prove to be statistically related to postoperative hearing. The variations in morphology and latency of CNAPs are discussed in relation to the mechanisms of iatrogenic injury. PMID- 9419087 TI - Thermal imaging of the temporal bone in CO2 laser surgery: an experimental model. AB - The unique properties of lasers create an enormous potential for specific treatment of chronic ear disease. Despite the widespread acceptance and use of the laser, however, a complete understanding of the time- and space-dependent temperature distribution in otic capsule bone immediately after pulsed laser exposure has not been elucidated. Using a liquid nitrogen-cooled mercury-cadmium telluride infrared detector, the temperature distribution in human cadaveric otic capsule bone was determined immediately after pulsed (100 msec) carbon dioxide laser exposure (0.3 to 4.0 W; 200 microm spot diameter). The time- and space dependent temperature increases and thermal diffusion were determined as a function of the laser power density and were found to vary linearly. PMID- 9419088 TI - The effect of acidic fibroblast growth factor and live yeast cell derivative on tympanic membrane regeneration in a rat model. AB - The current treatment of choice for chronic tympanic membrane perforations is surgery. Recent studies using various polypeptide growth factors to accelerate closure of tympanic membrane perforations in model systems have produced mixed results. This study evaluates the effect of acidic fibroblast growth factor (AFGF) and live yeast cell derivative (LYCD) on the rate of healing of acute tympanic membrane perforations in a rat model. Thirty-seven rats had both ears separately randomized in a blinded fashion to receive AFGF in one of three concentrations, LYCD, or a control solution. The rats initially underwent subtotal removal of the tympanic membranes bilaterally. Solutions were applied to the randomized ears daily for 3 days, starting at the time of the surgical perforation. The ears were photographed every 3 to 8 days for 35 days. The photographs were digitally scanned and a computer analysis was used to calculate the percentage of residual perforation. No significant difference in the rate of healing was observed for ears treated with AFGF or LYCD versus the controls. Given the potential advantages of medical treatment of tympanic membrane perforations and the established efficacy of growth factors in other model systems, however, further research is warranted. PMID- 9419089 TI - Expert opinion on the diagnosis of acoustic tumors. AB - Ideally, clinicians recommend diagnostic tests when the patient's risk of disease is sufficient to justify putting numerous similar patients through the morbidity required to diagnose disease in one patient. In the case of acoustic tumor diagnosis, there are few published data available to the clinician to help assess risk in an individual patient. The purpose of this study was to obtain information by an opinion poll of a group of experts. We used the Delphi method to poll clinicians trained at the House Ear Clinic. We asked these experts 20 questions related to acoustic tumor diagnosis. Some of the expert opinion presented herein is the only data related to acoustic tumor diagnosis available to clinicians. These data are a first step in elevation of decision-making for tumor diagnosis above the level of speculation. However, the experts' responses displayed a pattern of inaccuracy that limits the clinical application of their opinion. Exposing this pattern was instructive for identifying desirable features of protocols for diagnosing tumors. We recommend that protocols not depend on clinicians estimating probability of tumor. Instead, protocols may list specific findings, such as unilateral distortion on the telephone, to indicate, when present, that the risk of tumor is sufficient to order a diagnostic test. PMID- 9419090 TI - Value of preoperative prothrombin time/partial thromboplastin time as a predictor of postoperative hemorrhage in pediatric patients undergoing tonsillectomy. AB - OBJECTIVE: Hemorrhage after tonsillectomy is a potentially lethal complication. Preoperative assessment consisting of prothrombin time (PT) and activated partial thromboplastin time (PTT) has been used to identify patients at risk for hemorrhage after tonsillectomy and adenoidectomy. We sought to assess the value of PT/PTT screening as a predictor of posttonsillectomy hemorrhage. DESIGN: A retrospective chart review was carried out with a minimum of 1 month follow-up. SETTING: Tertiary academic referral center. PATIENTS: Between January 1992 and June 1995, 382 patients undergoing tonsillectomy were examined; 339 patients with a minimum of 1 month follow-up were reviewed for this study. MAIN OUTCOME MEASURE: Normal and prolonged PT/PTT values were examined. Bleeding in the intraoperative, immediate postoperative, and delayed phases of healing was examined. RESULTS: Two-hundred and twenty-two patients had normal PT/PTT, 39 had prolonged PT/PTT, and 78 had no preoperative studies performed. Bleeding occurred in 2.7%, 2.6%, and 3.3%, respectively, of patients. Eight patients had positive family histories of bleeding tendencies. One patient (12.5%) with a normal PT/PTT experienced a delayed posttonsillectomy bleed. Of 39 patients with abnormal coagulation studies, 30 were borderline elevations with no repeat studies done; one patient experienced postoperative hemorrhage. Nine abnormal results were repeated; three returned to normal, three remained prolonged but underwent tonsillectomy with no intervention, and three received hematology consultations. One patient had lupus anticoagulant, one had Hageman Factor deficiency, and one was cleared for surgery with no diagnosis. All patients underwent tonsillectomy with no episodes of postoperative bleeding. CONCLUSIONS: Preoperative PT/PTT provides no additional information than does a bleeding history for the general pediatric population undergoing tonsillectomy. This should only be done in selective cases where warranted by history. PMID- 9419091 TI - Sinus mucoceles: is marsupialization enough? AB - Traditional teaching in the United States has emphasized the need for complete removal of sinus mucoceles to achieve a cure. In Europe, however, many rhinologic surgeons have been treating sinus mucoceles by draining and marsupializing them. We present our experience with the treatment of 16 patients with sinus mucoceles. This series includes nine frontal, two ethmoid, two sphenoid, one sphenoethmoid, and two maxillary sinus mucoceles. All patients were treated transnasally under telescopic control. All mucoceles were marsupialized, and 11 were stented. Intraoperative transillumination as well as intraoperative lateral x rays (for sphenoethmoid lesions) and anteroposterior x rays (for frontal mucoceles) were used to ensure complete marsupialization of the lesion and to confirm proper placement of the stent. There were no complications associated with the procedure. Follow-up periods ranged from 8 to 62 months (median, 32 months). No evidence of recurrent mucocele has been seen in any of the patients. PMID- 9419092 TI - Microscopic tonsillectomy: a double-blind randomized trial. AB - OBJECTIVE: To evaluate microsurgical bipolar cautery tonsillectomy (TEmic) by comparing it with traditional blunt dissection tonsillectomy (TEtrad). DESIGN: A double-blind prospective randomized trial with stratification in two age groups. PATIENTS: 200 consecutive patients undergoing tonsillectomy for tonsillar hypertrophy, or recurrent or chronic tonsillitis. OUTCOME MEASURES: Duration of surgery, intraoperative bleeding, daily postoperative pain and otalgia, postoperative bleeding episodes. METHODS: Duration of surgery and operative bleeding were evaluated by the anesthesiologist. The patients were instructed to record daily pain and otalgia. Final postoperative evaluation was done by a different physician, blinded to the surgical technique. RESULTS: Mean intraoperative bleeding was 12 ml for TEmic and 36 ml for TEtrad (P < 0.001). Mean duration of surgery was 37 minutes for TEmic and 36 minutes for TEtrad (NS). Otalgia was present in 41% of TEmic patients and 69% of TEtrad patients (p < 0.001). Daily postoperative pain was lower in the TEmic group than it was in the TEtrad group for the entire study period (10 days). Postoperative hemorrhage was present in three TEmic patients (3%) and in eight TEtrad patients (8%), a difference that did not reach significance (p > 0.1). CONCLUSION: Microsurgical bipolar cautery tonsillectomy compares favorably with traditional techniques in terms of intraoperative bleeding, postoperative pain, otalgia, and hemorrhage. This technique combines the hemostatic advantage of cautery dissection, the excellent visualization achieved by a microscope, and, with the use of a video, greatly improves the physician's ability to teach how to perform a tonsillectomy. PMID- 9419093 TI - Obstructive sleep apnea surgery: risk management and complications. AB - BACKGROUND: Hypoxemia, hypertension, airway obstruction, and death have been associated with surgery for obstructive sleep apnea syndrome (OSAS). Patient analysis was undertaken to identify potential factors that could affect risk management outcome. METHODS: One hundred eighty-two consecutively treated patients with OSAS undergoing 210 procedures were evaluated. Fifty-four factors were analyzed. RESULTS: Group characteristics included a mean age of 48.2 years, a mean respiratory disturbance index of 42.3, and a mean low oxyhemoglobin desaturation (LSAT) of 77.5%. Surgery included a combination of uvulopalatopharyngoplasty (162 patients; 77%) and maxillofacial procedures (173 patients; 82%). Patients with a respiratory disturbance index greater than 40 and an LSAT less than 80% (117 patients; 64%) were maintained on nasal continuous positive airway pressure. Thirty-nine patients (18.6% had difficult intubations. There was a positive correlation (p > 0.001) of difficult intubations, neck circumference (> 45.6 cm) and skeletal deficiency (Sella-Nasion-Point B < 75 degrees). All tubes were removed with the patient awake in the operating room with two transient episodes of airway obstruction. One hundred forty-eight of the patients (70.5%) required postoperative intravenous antihypertensive medications. Patients with a preoperative history of hypertension had a significantly increased risk (p > 0.01) of requiring intraoperative and postoperative intravenous antihypertensive medications. The mean hospital stay was 2.2 days (SD +/- 0.9). Analgesia was achieved with intravenous morphine sulfate or meperidine HCl (intensive care unit) and oral oxycodone (non-intensive care unit). There were no significant oxyhemoglobin desaturations, irrespective of severity of OSAS or obesity (mean LSAT day 1, 94.8% (SD +/- 2.4); mean LSAT day 2, 95.5% (SD +/- 1.6)). Complications included postoperative bleeding (n = 4), infection (n = 5), seroma (n = 3), arrhythmia (n = 4), angina (n = 1), and loss of skeletal fixation (n = 1). CONCLUSION: Intraoperative airway risks can be reduced by use of fiberoptic intubation in patients with increased neck circumference and skeletal deficiency. Patients with OSAS are at a significantly increased risk for hypertension. Nasal continuous positive airway pressure eliminated the postoperative risk of hypoxemia, which allowed the use of adequate parenteral or oral analgesics. PMID- 9419094 TI - Use of a screening RAST in a large neuro-otologic practice. AB - Evidence in the literature emphasizes the role of the immune system in disorders of the inner ear and eustachian tube. We initially investigated the presence of inhalant allergy in selected patients seen for otologic problems by means of a screening radioallergosorbent test (RAST), using either a microscreen or a limited antigen panel. This study analyzed the results of tests performed over a 2-year period on 186 patients seen by one of us (WLM) for treatment of vertigo (66%), tinnitus (63%), hearing loss (49%), aural fullness (48%), Meniere's quadrad (27%), balance disturbance other than true vertigo (21%), and eustachian tube dysfunction (4%). We found an incidence of immunoglobulin E-mediated hypersensitivity of nearly 40% in a patient population selected solely for neuro otologic symptoms and not for sinonasal symptoms. This figure is more than double that quoted for the general population. We also found a surprisingly high incidence of mold antigen atopy in this selected population. Allergy can contribute to a number of otologic symptoms, including eustachian tube dysfunction, vertigo, tinnitus, hearing loss, aural fullness, and nonspecific balance disturbance. Allergy also has been emphasized as an etiologic factor in a portion of patients diagnosed with Meniere's syndrome. A screening RAST, combined with clinical evaluation, appears to be an excellent tool for evaluating these patients for inhalant allergy as part of a comprehensive workup. PMID- 9419095 TI - Use of color Doppler flow imaging for preoperative assessment in fibular osteoseptocutaneous free tissue transfer. AB - Fibular osteocutaneous free tissue transfer represents state-of-the-art reconstruction of the oromandibular complex after oncologic resection. Because the blood supply of the fibular flap is based on the peroneal artery and venae comitantes, it is essential to determine preoperatively whether adequate perfusion to the donor extremity will persist after sacrifice of the peroneal pedicle. Vascular variations or peripheral arterial occlusive disease may exist whereby sacrifice of peroneal vessels can cause ischemia of the lower leg and foot. Therefore, angiography of the lower extremity and magnetic resonance angiography have been advocated to determine the arterial supply to the lower leg. Though accurate, these examinations carry risk for morbidity, and both are expensive. Color Doppler flow imaging is a noninvasive ultrasound examination used to measure antegrade and retrograde blood flow. We prospectively evaluated color Doppler flow imaging as a means of preoperative evaluation of the lower extremity for candidates for fibular flaps. Ability to image peroneal vessels and to determine collateral and distal perfusion were evaluated. The patients were compared with a control group of patients with peripheral arterial occlusive disease. Color Doppler flow imaging provided accurate hemodynamic information for all patients and allowed successful fibular transfer for all patients who received free flaps. The modality is comparable in accuracy with the angiography of the lower extremity and magnetic resonance angiography and has the advantages of low cost and no morbidity. PMID- 9419096 TI - Bone graft incorporation after cortical perforations of the host bed. AB - A fundamental issue in onlay bone graft persistence is the unpredictable extent of incorporation and volumetric maintenance of the graft. The purpose of this study was to evaluate the effects on integration of onlays, with either their cancellous or cortical portion facing toward the host bed, positioned over cortical perforations at the recipient site. Tibial or femoral unicortical bone grafts were harvested from isogeneic donors and positioned subperiostally on each tibia of 22 adult Lewis rats. On the experimental side, the recipient outer cortical bone surface received multiple perforations, 0.25 mm in diameter. The contralateral side served as a control (no cortical perforations). The findings were assessed after 4 and 20 weeks using routine histologic and immunohistochemistry techniques. Cortical perforations induced a migration of the recipient bone marrow into the graft as well as a reduced size diminution. More cortical bone remodeling and marginal lamellar bone apposition were observed after orientating the cortical portion of the graft toward the recipient site. These observations may be useful clinically to improve long-term success after autogeneic bone grafting. PMID- 9419097 TI - A cadaveric study of the motor nerves to the levator scapulae muscle. AB - Understanding the surgical anatomic relationships of the motor nerves to the levator scapulae muscle is imperative for reducing postoperative shoulder dysfunction in patients undergoing neck dissection. To elucidate this relevant anatomy, cervical (C3, C4) and brachial (C5 via dorsal scapular nerve) plexi contributions to the levator scapulae were assessed with respect to posterior triangle landmarks in 37 human cadaveric necks. An average of approximately 2 (actual 1.92) nerves from the cervical plexus (range 1 to 4 nerves) emerged from beneath the posterior border of the sternocleidomastoid muscle in a cephalad to caudad progression to enter the posterior triangle of the neck on their way to innervating the levator scapulae. These cervical plexus contributions exhibited a fairly regular relationship to the emergence of cranial nerve XI and the punctum nervosum along the posterior border of the sternocleidomastoid muscle. After emerging from the posterior border of the sternocleidomastoid to enter the posterior triangle of the neck, cervical plexus contributions to the levator scapulae traveled for a variable distance posteriorly and inferiorly, sometimes branching or coming together. Ultimately these nerves crossed the anterior border of the levator scapulae as 1 to 3 nerves (average 1.94) in a regular superior to inferior progression. The dorsal scapular nerve from the brachial plexus exhibited highly variable anatomic relations in the inferior aspect of the posterior triangle, and was found to penetrate or give branches to the levator scapulae in only 11 of 35 neck specimens. We have found that the levator scapulae receives predictable motor supply from the cervical plexus. Our data elucidate surgical anatomy useful to head and neck surgeons. PMID- 9419098 TI - Endoscopic laser resection of supraglottic carcinoma. AB - Forty-six patients with infiltrating supraglottic carcinoma were treated prospectively for cure between 1986 and 1992 with transoral laser resection of the primary. Nine primaries were classified as T1, and 37 as T2. Thirty-three had staged unilateral or bilateral neck dissections, and 16 had postoperative radiotherapy. All patients were followed up for 2 to 8 years unless they died. Of the 46 patients, 33 are alive without disease, 8 died with disease, and 5 died of intercurrent disease. Among the 8 patients who died with disease, 4 had uncontrollable local or regional recurrences, and 4 had distant metastases but were free of local or regional recurrence. Calculated overall survival was 59% and adjusted survival was 72% after 5 years. Four patients had tracheostomies perioperatively, and 2 required temporary postoperative tracheostomies. The remaining 40 patients needed no artificial airway other than orotracheal intubation for the endolaryngeal intervention. Thirty-seven patients relearned undisturbed deglutition within 2 weeks from surgery, and 4, within 4 weeks. However, 5 (10.9%) patients failed to relearn swallowing and consecutively underwent "completion" total laryngectomy. Among them were the two patients who had previously had unsuccessful surgical or radiologic treatment of their primaries and the patient with a history of oral cavity carcinoma. This study confirms that transoral laser resection can effectively control early supraglottic carcinoma. Tracheostomies are not routinely required, and phonatory function is not compromised. However, transoral laser resection could not steadily preserve undisturbed deglutition in the patients included in this study. Patients with histories of unsuccessful attempts of other larynx-sparing therapeutic modalities or of previous major head and neck interventions were not successfully managed with transoral laser resection in this series. Criteria for patient selection remain to be established. PMID- 9419099 TI - Undifferentiated carcinoma of nasopharyngeal type of the laryngopharyngeal region. AB - Undifferentiated carcinoma of nasopharyngeal type (lymphoepithelioma) is an extremely rare malignancy in the laryngohypopharyngeal region. We found reports of only 13 such tumors in the English language literature. We present the findings of four additional cases, one hypopharyngeal and three laryngeal in origin. The three laryngeal tumors were characterized by submucosal spread. The tumors were classified T3 (2x) and T4 (2x) with cervical lymph node metastases at initial presentation in all cases. In three of our four cases the Epstein-Barr virus was demonstrated by the Epstein-Barr virus-encoded RNAs in situ hybridization. PMID- 9419100 TI - High-resolution ultrasound associated with aspiration biopsy in the follow-up of patients with differentiated thyroid cancer. AB - OBJECTIVE: To assess the value of ultrasound in the follow-up of patients undergoing surgery for differentiated thyroid carcinoma. SUBJECTS: The study included 89 patients (70 women and 19 men) with differentiated thyroid carcinoma (76 papillary and 13 follicular cancer). METHODS: High-frequency ultrasound (US) was used in the evaluation of 89 subjects who underwent surgery for thyroid carcinoma. Fine-needle aspiration was performed in cases with positive US. In addition, determinations of thyroglobulin (Tgb) in serum, scintigraphy with (131)I, and cervical palpation were evaluated. We determined sensitivity, specificity, and overall accuracy for each of these diagnostic methods. RESULTS: Ultrasonography was positive in 22 subjects, 16 in the nodal area and 6 in the thyroid bed. Twenty-two subjects received fine-needle aspiration with US control; 13 (59%) of 22 were positive for cancer. The results of the US for detecting neoplastic disease showed a sensitivity of 65%, specificity of 86%, and overall accuracy of 82%. The overall accuracy for scintigraphy was 88% and for Tgb, 91%. CONCLUSION: We concluded that US can be included in the follow-up protocol for patients undergoing surgery for differentiated cancer of thyroid, as a valuable tool to localize the recurrence. This technique is particularly useful in the evaluation of patients who are found to have elevated Tgb levels. PMID- 9419101 TI - Electronic access to tinnitus data: the Oregon Tinnitus Data Archive. AB - The recently published Oregon Tinnitus Data Archive can be viewed by anyone with Internet access and World Wide Web browser software. A dynamically changing document to be amplified and upgraded in future versions, at present the Archive summarizes records from 1630 patients with tinnitus seen between 1982 and 1992 at the Tinnitus Clinic of the Oregon Health Sciences University. The Archive has many features of standard journal publications but also has added advantages of extensive appendixes and other background information, as well as the ease of use and flexibility offered by hypertext documents. The data in the Archive can be used for a variety of clinical and research purposes such as development of prevalence estimates for medical conditions or etiologic circumstances associated with tinnitus, testing of clinical impressions and other hypotheses, and efforts to identify risk factors and possible causal agents for tinnitus. PMID- 9419102 TI - Tinnitus suppression in cochlear implant patients. AB - Tinnitus is sometimes suppressed by cochlear implantation. Tinnitus conditions were examined in 60 adult patients who underwent cochlear implantation in 1990 or later. Before surgery, 90% of these patients reported some type of tinnitus. The patients completed a questionnaire evaluating the loudness and duration of the tinnitus, immediately after the first electrical stimulation and 2 months later. The loudness of tinnitus was suppressed in 65% of the patients at first evaluation. Two months later, the tinnitus was suppressed in 93% of the patients. As for the duration of tinnitus, at first evaluation the tinnitus duration was suppressed in 41% of the patients. Two months later, the duration of tinnitus was suppressed in 61% of the patients. PMID- 9419103 TI - Test-retest reliability of vestibular autorotation testing in healthy subjects. AB - Vestibulo-ocular reflex rotational chair testing in the high-frequency range is seldom performed because it requires specialized and powerful systems. But today a new method of sweep-frequency vestibulo-ocular reflex testing, the Vestibular Autorotation Test system (Western Systems Research, Inc., Pasadena, Calif.), based on active head movements increasing from 2 to 6 Hz, is available on the market. The goal of this study was to evaluate the test-retest variability of this test in healthy subjects. Twelve young adults (22 to 42 years old) without any history of auditory or vestibular dysfunction were included in the study. Subjects underwent five tests under standardized conditions with a 1-week interval. Each test consisted of three measurements of the gain and phase of the vestibulo-ocular reflex in the horizontal and vertical planes. Statistical analysis shows that the test-retest reliability of the Vestibular Autorotation Test is poor. Therefore this method cannot be used routinely to evaluate precise vestibulo-ocular reflex anomalies. PMID- 9419104 TI - Laryngeal melanosis: report of four cases and literature review. AB - OBJECTIVE: Laryngeal melanosis is a rare condition defined by the presence of melanocytes within the laryngeal epithelial lining. Our aims were (1) to review our cases together with those in the literature, and (2) to determine whether melanocyte incidence is increased with exposure to irritant stimuli such as tobacco. METHODS: A retrospective study of all cases diagnosed with laryngeal melanosis in our hospital from January 1, 1990, to December 31, 1996, was accomplished. To determine the melanocyte incidence in the normal larynx as well as the influence of tobacco in development of laryngeal melanosis, 16 age-matched controls, 8 of whom were smokers and 8 of whom were not, were chosen, and a histochemical and immunohistochemical study was performed. The following antibodies were used: S-100 protein, CD1a, and HMB-45. A comparative study of the melanocyte incidence between patients with laryngeal melanosis and the controls was carried out. Also, a comparative study between smoking and nonsmoking patients was performed. RESULTS: Laryngeal melanosis was diagnosed in 4 patients at our hospital during this period of time. In the comparative study, the number of melanocytes in the 4 patients with laryngeal melanosis was higher than in the 8 smoking (p < 0.01, Mann-Whitney U test) and 8 nonsmoking (p < 0.01) controls, and there was a trend toward a higher number of melanocytes in the 8 smoking patients than in the 8 nonsmoking (p = 0.064) controls. CONCLUSIONS: Laryngeal melanosis was more frequent in smoking men older than 50 years. Our observations underline the association of LM with larynx carcinoma and its relation to a stimulus such as tobacco. In fact, we have found activated melanocytes in our cases of laryngeal melanosis. They were identified by immunoreactivity for HMB 45. PMID- 9419105 TI - Solitary neurofibroma of the larynx. PMID- 9419106 TI - Eosinophilic ulcer of the tongue. PMID- 9419107 TI - Valsalva-induced cervical thymic cyst. PMID- 9419108 TI - Management of odontogenic myxoma of the maxilla. PMID- 9419109 TI - Collision tumors at the cerebellopontine angle: case report with literature review. PMID- 9419110 TI - Angiolipoma of internal auditory canal presenting repeated sudden hearing loss. PMID- 9419111 TI - Posttraumatic synostosis of the cervical spine to the thyroid cartilage presenting as dysphagia. PMID- 9419112 TI - Coronoid osteochondroma of the mandible: transzygomatic access and autogenous bony reconstruction. PMID- 9419113 TI - Invasive Pseudallescheria boydii fungal infection of the temporal bone. PMID- 9419114 TI - Alveolar soft part sarcoma: report of a case occurring in the larynx. PMID- 9419115 TI - Primary osteoma cutis. PMID- 9419116 TI - Lingual thyroid and neoplastic change: a review of the literature and description of a case. PMID- 9419117 TI - Impaired ciliary clearance from tracheopathia osteoplastica of the upper respiratory tract. PMID- 9419118 TI - Split pectoralis major flap for massive tracheoesophageal fistula. PMID- 9419119 TI - Cementifying fibroma of the frontoethmoid complex. PMID- 9419120 TI - A rare case of nasopharyngeal angiofibroma in a pregnant woman. PMID- 9419121 TI - Complete facial paralysis as a result of parotid abscess. PMID- 9419122 TI - Papillary thyroid carcinoma presenting with massive angioinvasion of the great vessels of the neck and chest. PMID- 9419123 TI - Electromyographic examination of patients with unilateral cortical facial paralysis. PMID- 9419124 TI - Ancient schwannoma of the posterolateral pharynx: a benign lesion commonly mistaken for sarcoma. PMID- 9419125 TI - Interesting presentation of spinal muscular atrophy: cricoarytenoid joint fixation. PMID- 9419126 TI - Mycobacterium fortuitum otitis media. PMID- 9419127 TI - Lateral sinus thrombophlebitis. PMID- 9419128 TI - Transconjunctival endoscopic orbital decompression. PMID- 9419129 TI - An unusual cause of eustachian tube dysfunction. PMID- 9419130 TI - Is surgical excision of facial nerve schwannomas always indicated? PMID- 9419131 TI - Complication of maxillary sinus Foley balloon placement for orbital floor support. PMID- 9419132 TI - Osteoma of the malleus. PMID- 9419133 TI - Mandibular aneurysmal bone cyst associated with fibrous dysplasia. PMID- 9419134 TI - Magnetic resonance imaging in carotidynia. PMID- 9419135 TI - Mycobacterium avium complex infection of the paranasal sinuses. PMID- 9419136 TI - Positron emission tomography imaging of a branchial cleft cyst in a 45-year-old man. PMID- 9419137 TI - Dermatitis herpetiformis: an immunologically mediated form of gluten-sensitive enteropathy with head and neck manifestations. PMID- 9419138 TI - Temporomandibular joint prolapse after tympanoplasty. PMID- 9419139 TI - Ludwig's angina: improved treatment. PMID- 9419140 TI - Ketorlac-induced status asthmaticus after endoscopic sinus surgery in a patient with Samter's triad. PMID- 9419141 TI - Caffeine withdrawal after head and neck surgery. PMID- 9419143 TI - Isolated neurosarcoidosis presenting as anosmia and visual changes. PMID- 9419142 TI - An unusual presentation of the sternocleidomastoid tumor of infancy. PMID- 9419144 TI - Tracheotomy removal after early mandibular advancement in patients with pediatric craniofacial syndrome. PMID- 9419145 TI - Chiari-I malformation presenting as vocal cord paralysis in the adult. PMID- 9419146 TI - Inner ear membrane ruptures demonstrated with keratin immunohistochemistry. PMID- 9419147 TI - Traumatic internal carotid artery thrombosis after impalement injury. PMID- 9419148 TI - Fibromyxoma of the temporal bone. PMID- 9419149 TI - Non-Hodgkin's lymphoma of the middle ear cleft. PMID- 9419150 TI - Cervical sympathetic schwannoma: a case report and review of the English literature. PMID- 9419151 TI - Epithelioid sarcoma of the neck: a rare tumor mimicking metastatic carcinoma from an unknown primary. PMID- 9419152 TI - Wound botulism: a clinical experience. PMID- 9419153 TI - Orbitoethmoid osteoma: case report of an uncommon presentation of an uncommon tumor. PMID- 9419154 TI - X-inactivation and cytogenetic studies in a family with sensorineural hearing loss and Turner syndrome. PMID- 9419155 TI - Chronic sinusitis with acquired immunoglobulin A (IgA) deficiency after bone marrow transplantation. PMID- 9419156 TI - Infiltrating papillary carcinoma of the thyroid with macroscopic extension into the internal jugular vein. PMID- 9419157 TI - Videostroboscopic findings in laryngeal tuberculosis. PMID- 9419158 TI - Lipoblastoma in the parotid gland of an infant. PMID- 9419159 TI - alpha-Interferon toxicity after septoplasty: a diagnostic dilemma. PMID- 9419160 TI - Paraneoplastic acral hyperkeratosis: initial sign of laryngeal neoplasia. PMID- 9419161 TI - Multiple cystic parathyroid adenoma in a geriatric patient with primary hyperparathyroidism. PMID- 9419162 TI - Summary of the International Conference on Emerging Infectious Diseases US-Japan Cooperative Medical Science Program, Bangkok, 1997. PMID- 9419163 TI - HLA-DR class II associations with rubella vaccine-induced joint manifestations. AB - HLA class II (HLA-DR) frequencies were examined in relation to incidence of acute arthralgia or arthritis in 283 white women who had received RA27/3 rubella vaccine (n = 146) or placebo (n = 137) postpartum. Leukocyte DNA was molecularly typed for HLA-DRB1 gene expression. Univariate analysis revealed higher frequencies of DR2 (odds ratio [OR], 4.8; 95% confidence interval [CI], 1.2-18.8) and DR5 (OR, 7.5; 95% CI, 1.5-37.5) but lower frequencies of DR4 (OR, 2.3; 95% CI, 1.1-4.9) and DR6 (OR, 2.8; 95% CI, 1.4-5.8), in rubella vaccinees compared with placebo recipients with arthropathy. Logistic regression modelling of DR, treatment, age, time postpartum, and arthropathy revealed that the odds of developing arthropathy was 1.9 times greater (95% CI, 1.07-3.44) after rubella vaccine than placebo. Risk for arthropathy (regardless of rubella vaccination) was also influenced by DR interactions: odds were 8 times greater in individuals with both DR1 and DR4 (95% CI, 1.45-44.02) and 7.1 times greater with both DR4 and DR6 present (95% CI, 1.85-27.52), suggesting that coexpression of these specificities may predispose to postpartum arthropathy. PMID- 9419164 TI - Hospitalizations associated with rotavirus diarrhea in the United States, 1993 through 1995: surveillance based on the new ICD-9-CM rotavirus-specific diagnostic code. AB - The introduction of a specific International Classification of Diseases code for rotavirus diarrhea in 1992 prompted examination of the National Hospital Discharge Survey (NHDS) for trends in rotavirus-associated hospitalizations among US children aged 1 month through 4 years. During 1993-1995, 13.5% of hospitalizations were associated with diarrhea (n = 162,478/year). Rotavirus was the most common pathogen identified, coded in 16.5% of diarrhea cases (n = 26,798/year), and increased from 13.3% in 1993 to 18.9% in 1995. The age distribution and seasonality of hospitalizations of presumed noninfectious and viral etiology resembled those associated with rotavirus. Rotavirus was reported as a cause of diarrhea more frequently by hospitals that were large (> or =100 beds), proprietary-owned, or in the West/Midwest. Although these findings suggest incomplete detection of rotavirus diarrhea cases, the large number of rotavirus associated hospitalizations underscores the need for vaccines and indicates that NHDS data could be used to monitor the impact of a US rotavirus immunization program. PMID- 9419165 TI - Mucosally transmitted feline immunodeficiency virus induces a CD8+ antiviral response that correlates with reduction of cell-associated virus. AB - Intravaginal inoculation of cats with feline immunodeficiency virus (FIV) results in acute systemic infection accompanied by a strong CD8+ immune response that inhibits viral replication. CD8+ anti-FIV activity, revealed by increased FIV replication in peripheral blood mononuclear cells (PBMC) depleted of CD8+ lymphocytes, was detected by 6 weeks after inoculation and correlated with reduced PBMC-associated virus at 12, 16, and 32 weeks after inoculation. Some cats with strong CD8+ anti-FIV activity during acute infection did not seroconvert and yielded no evidence of FIV infection at later times. These data suggest that CD8+ immunity may play a major role in eliminating virus during primary transmucosal FIV infection and may down-regulate viral replication during asymptomatic infection. PMID- 9419166 TI - Induction of Th2 cytokine expression for p27-specific IgA B cell responses after targeted lymph node immunization with simian immunodeficiency virus antigens in rhesus macaques. AB - To determine if there is an association between the isotype of simian immunodeficiency virus (SIV)-specific B cell responses and the profile of Th1 and Th2 cytokine expression, rhesus macaques were immunized with SIV antigens via the iliac lymph nodes, using a targeted lymph node (TLN) immunization procedure. When CD4+ T cells purified from antigen-stimulated peripheral blood mononuclear cells were analyzed, the levels of Th2 cytokine production were gradually increased after the second and third immunizations. However, interferon-gamma production did not change. Analysis of SIV-specific B cell responses revealed that the main isotype was IgG after the second and third immunizations. In addition, a peak of SIV-specific IgA B cell responses was noted following the third immunization. These findings suggest that the induction of Th2 type responses in TLN-immunized rhesus macaques reflects the sequence of initial induction of SIV-specific IgG producing cells followed by IgA-secreting cells. PMID- 9419167 TI - Cell-free human immunodeficiency virus type 1 in breast milk. AB - Breast-feeding may be an important route of human immunodeficiency virus type 1 (HIV-1) vertical transmission in settings where it is routinely practiced. To define the prevalence and quantity of HIV-1 in cell-free breast milk, samples from HIV-1-seropositive women were analyzed by quantitative competitive reverse transcription-polymerase chain reaction (QC-RT-PCR). HIV-1 RNA was detected in 29 (39%) of 75 specimens tested. Of these 29 specimens, 16 (55%) had levels that were near the detection limit of the assay (240 copies/mL), while 6 (21%) had >900 copies/mL. The maximum concentration of HIV-1 RNA detected was 8100 copies/mL. The prevalence of cell-free HIV-1 was higher in mature milk (47%) than in colostrum (27%, P = 0.1). Because mature milk is consumed in large quantities, these data suggest that cell-free HIV-1 in breast milk may contribute to vertical transmission of HIV-1. PMID- 9419168 TI - Use of changes in plasma levels of human immunodeficiency virus type 1 RNA to assess the clinical benefit of antiretroviral therapy. AB - Data from 1330 human immunodeficiency virus type 1 (HIV-1)-infected patients enrolled in seven antiretroviral treatment trials were analyzed to characterize the clinical benefit of treatment-mediated reductions in plasma HIV-1 RNA levels. The risk of a new AIDS-defining event or death was reduced proportionally to the magnitude of the reduction of the HIV-1 RNA level during the first 6 months of therapy. Pretherapy HIV-1 RNA levels were prognostic independently of on-therapy levels. In addition, the reduction in risk associated with any given reduction of the level of HIV-1 RNA did not vary by pretherapy level. Having either a reduction in HIV-1 RNA level or an increase in CD4+ lymphocyte count, or both, was associated with a delay in clinical disease progression. This indicates that patient prognosis should be assessed using both HIV-1 RNA and CD4+ lymphocyte responses to therapy. PMID- 9419169 TI - A randomized, double-blind trial of valaciclovir prophylaxis for cytomegalovirus disease in patients with advanced human immunodeficiency virus infection. AIDS Clinical Trials Group Protocol 204/Glaxo Wellcome 123-014 International CMV Prophylaxis Study Group. AB - Cytomegalovirus (CMV) disease is a common complication of advanced human immunodeficiency virus (HIV) infection. Administration of oral valaciclovir, a valine ester of acyclovir, achieves sufficient plasma acyclovir levels to inhibit many clinical isolates. Acyclovir has been associated with enhanced survival in AIDS but not with CMV disease prevention. CMV-seropositive patients (1227) with CD4 cell counts <100/mm3 were enrolled in a randomized, double-blind trial. Valaciclovir, 8 g/day, was compared with acyclovir, 3.2 or 0.8 g/day, for CMV prevention; all three arms were compared for survival. The confirmed CMV disease rate was 11.7% among valaciclovir recipients and 17.5% in the pooled acyclovir arms, a 33% reduction in risk. Time to confirmed CMV disease was significantly longer for the valaciclovir group (P = .03). A trend toward earlier mortality for valaciclovir recipients was seen (P = .06). Toxicity and earlier medication discontinuation were more common in this group. Valaciclovir significantly reduces the risk of CMV disease. Further exploration of a better-tolerated dose is warranted. PMID- 9419170 TI - The effect of valaciclovir on cytomegalovirus viremia and viruria detected by polymerase chain reaction in patients with advanced human immunodeficiency virus disease. AIDS Clinical Trials Group Protocol 204/Glaxo Wellcome 123-014 International CMV Prophylaxis Study Group. AB - Samples of blood and urine were collected at baseline, week 4, and week 8 and then every 8 weeks from 310 patients entering a controlled trial of prophylaxis with valaciclovir versus acyclovir. Samples were tested under code by polymerase chain reaction (PCR) in one laboratory. The median number of samples collected from each patient was 5 for blood (range, 0-15) and 5 for urine (range, 0-15). Both baseline PCR viremia and PCR viruria were significantly associated with future cytomegalovirus (CMV) disease (P = .002 and P = .02, respectively). The greatest effect of valaciclovir on CMV disease was seen in patients who were PCR positive in blood at baseline (P = .002), although a significant effect was also seen in those who were PCR-negative in urine (P = .02). Thus, PCR viremia provides prognostic information about CMV disease in AIDS patients, and valaciclovir showed activity as both a preemptive and prophylactic agent. PMID- 9419171 TI - Evidence of Nef truncation in human immunodeficiency virus type 2 infection. AB - Human immunodeficiency virus (HIV)-2 differs from HIV-1 in its relative lower transmissibility and pathogenicity. To understand the virologic basis of these differences, the nef gene from HIV-2-seropositive persons was analyzed because of its importance for disease progression in the genetically related simian immunodeficiency virus (SIV[MAC]). Proviral nef sequences from 60 HIV-2-infected persons were amplified from peripheral blood lymphocytes, and nef open-reading frames were screened by a transcription and translation assay for the presence of full-length (32- to 36-kDa) or truncated (<32 kDa) Nef proteins. Overall, 6 (10%) of 60 persons had truncated Nef proteins; of these, 5 were among the 36 asymptomatic subjects (13.9%) and only 1 was among the 24 symptomatic subjects (4.2%) (P =.23). The results of this study document the presence of defective nef genes in HIV-2 infections with a prevalence higher than that previously seen in HIV-1-infected cohorts of long-term nonprogressors or patients with AIDS. PMID- 9419172 TI - Persistent infection by Helicobacter pylori down-modulates virus-specific CD8+ cytotoxic T cell response and prolongs viral infection. AB - To determine whether Helicobacter pylori infection affects clearance of a concomitant viral infection and cytotoxic T lymphocyte (CTL) and cytokine response to that infection, H. pylori-infected BALB/c mice were challenged with a recombinant vaccinia virus expressing human immunodeficiency virus type 1 gp160. Two H. pylori strains, a colonizing clinical isolate (KS612) and an established standard noncolonizing strain (NCTC11637), were compared. Clearance of recombinant vaccinia virus was reduced in KS612-infected mice compared with NCTC11637-infected and control mice. As a potential mechanism, in contrast to control or NCTC11637-infected mice, the H. pylori clinical isolate KS612 diminished gp160-specific and vaccinia virus-specific CTL activity, even in the presence of exogenous interleukin-2. Furthermore, KS612-infected mice had reduced Th1 cytokine responses to gp120 in vitro compared with control or NCTC11637 infected mice. These results have implications for possible effects of prevalent H. pylori infection on other human diseases. PMID- 9419173 TI - Complement activation in patients with sepsis is in part mediated by C-reactive protein. AB - The involvement of C-reactive protein (CRP) in the activation of complement in patients with sepsis was investigated. In 104 patients with infections of varying severity, circulating levels of CRP-complement complexes, which are specific indicators for CRP-mediated complement activation, were assessed. Complement-CRP complexes were increased in almost all patients and correlated significantly with levels of C3a (r = .59; P < .001) and C-reactive protein (r = .76; P < .001). In addition, they correlated with levels of secretory phospholipase A2 (r = .59; P < .001). Levels of complement-CRP complexes in patients with a pneumococcal type of infection were similar to those in patients with other types of infections. Complement-CRP complexes were significantly higher in patients with shock (P = .01) and in patients who died (P = .03). These results demonstrate that part of the complement activation in patients with sepsis is independent from a direct interaction with microorganisms but rather results from an endogenous mechanism involving CRP. PMID- 9419174 TI - Enteroaggregative Escherichia coli produce intestinal inflammation and growth impairment and cause interleukin-8 release from intestinal epithelial cells. AB - Enteroaggregative E. coli (EAggEC) are emerging as an important cause of persistent diarrhea, especially in children in the developing world, yet the pathogenesis of EAggEC infection is poorly understood. In an ongoing prospective study of childhood diarrhea in an urban Brazilian slum, EAggEC are the leading cause of persistent diarrhea. Children from this study with EAggEC and persistent diarrhea had significant elevations in fecal lactoferrin, interleukin (IL)-8, and IL-1beta. Moreover, children with EAggEC without diarrhea had elevated fecal lactoferrin and IL-1beta concentrations. The children with EAggEC in their stool had significant growth impairment after their positive culture, regardless of the presence or absence of diarrhea. Finally, 2 EAggEC strains were shown to cause IL 8 release from Caco-2 cells, apparently via a novel heat-stable, high-molecular weight protein. These findings suggest that EAggEC may contribute to childhood malnutrition, trigger intestinal inflammation in vivo, and induce IL-8 secretion in vitro. PMID- 9419175 TI - papG alleles of Escherichia coli strains causing first-episode or recurrent acute cystitis in adult women. AB - The distribution of the three alleles of the P adhesin gene papG (classes I-III) was assessed among 74 Escherichia coli urine isolates from women with first episode or recurrent cystitis, and papG genotype was compared with clinical origin, O serogroup, agglutination of Gal(alpha1-4)Gal-coated latex beads and human or sheep erythrocytes, and hemolysin production. The class-III-only papG genotype (27% of strains) predominated over the I + III (3%) and II-only (7%) genotypes, irrespective of clinical category. In contrast to the class II papG allele, the class III allele was significantly concentrated in serogroups O6 and O18. Agglutination phenotypes corresponded significantly but incompletely with papG genotype, whereas hemolysin production and papG positivity were tightly correlated. These findings suggest that in acute cystitis in adult women, the class III papG allele predominates, confers distinctive agglutination phenotypes, and is restricted to specific E. coli lineages. PMID- 9419176 TI - Differences in N-acetylmuramyl-L-alanine amidase and lysozyme in serum and cerebrospinal fluid of patients with bacterial meningitis. AB - N-acetylmuramyl-L-alanine amidase (NAMLAA) specifically degrades peptidoglycan, a major component of bacterial cell walls. Lysozyme degrades peptidoglycan differently by hydrolyzing the aminosugar backbone of peptidoglycan. In another study, it was shown that the two enzymes act synergistically to inactivate the inflammatory properties of peptidoglycan. The presence of lysozyme and NAMLAA was determined in serum and cerebrospinal fluid (CSF) of patients with bacterial meningitis. High concentrations of lysozyme were found in CSF while, surprisingly, NAMLAA was not present. To explain this phenomenon, the degranulation pattern of neutrophils in CSF was compared with that of neutrophils from blood. Specific granules contain lysozyme and the azurophil granules contain both lysozyme and NAMLAA. CD66b expression on the cell surface, indicative for fusion of the specific granules with the cell membrane, was higher in CSF than in blood, while the marker for the azurophil granules was lower. PMID- 9419177 TI - Granulocyte-macrophage colony-stimulating factor induces activation and restores respiratory burst activity in monocytes from septic patients. AB - Monocyte activation in response to recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF) was examined in vitro in septic shock patients. These monocytes exhibited a greater respiratory burst activity than monocytes from healthy subjects; the response to secondary stimulation with bacterial stimuli was attenuated. GM-CSF restored the ability of monocytes to respond appropriately to secondary stimulation. Expression of certain integrin adhesion molecules, L-selectin, and Fcgamma receptors was increased on monocytes of septic shock patients; expression of CD11c was reduced. GM-CSF up-regulated integrin expression and decreased L-selectin, FcgammaRII, and FcgammaRIII expression. Septic patients exhibited greater biologic activity of monocyte tissue factor than did healthy subjects. Priming monocytes with GM-CSF accelerated tissue factor activation following stimulation with lipopolysaccharide and bacterial culture supernatant. Certain parameters of monocyte function may be restored by exposure to GM-CSF. This benefit may be offset by an increase in monocyte procoagulant activity. PMID- 9419178 TI - Effects of nitric oxide on chemotaxis and endotoxin-induced interleukin-8 production in human neutrophils. AB - The effects of nitric oxide (NO) on human neutrophil chemotactic responses and release of interleukin (IL)-8 was studied. Neutrophils exposed to chemoattractants (IL-8, FMLP, leukotriene B4, and C5a) failed to show increases in intracellular guanosine 3',5'-cyclic monophosphate (cGMP), an indicator of NO production. Although NO increased cGMP in neutrophils, neither of two NO donors (sodium nitroprusside and 3-morpholino-sydonimine) nor a NO synthase inhibitor (N omega-nitro-L-arginine) altered FMLP- or IL-8-elicited neutrophil chemotaxis (P > .25 for all). However, lipopolysaccharide-induced IL-8 production was increased in a dose-dependent manner by a combination of sodium nitroprusside and N acetylcysteine (P = .03) or by S-nitrosoglutathione (P = .004). NO-augmented IL-8 release was not reproduced by treating neutrophils with dibutyryl-cGMP. Up regulation of IL-8 release by NO was associated with increased IL-8 mRNA levels (P = .009). These data suggest that NO does not directly affect neutrophil chemotaxis but may indirectly alter chemotactic responses by increasing IL-8 production via a cGMP-independent pathway. PMID- 9419179 TI - Nonselective versus selective inhibition of inducible nitric oxide synthase in experimental endotoxic shock. AB - The effects of two nitric oxide synthase (NOS) inhibitors with different isoform selectivity were compared in a murine model of endotoxemia. Mice challenged with 70 mg/kg intraperitoneal (ip) lipopolysaccharide (LPS) were treated 6 h after LPS with either NG-gamma-L-arginine methyl ester (L-NAME, nonselective NOS inhibitor, 10-60 mg/kg), L-canavanine (selective inhibitor of inducible NOS, 50-300 mg/kg), or saline (0.2 mL) given ip. In a subset of mice, plasma concentrations of nitrate (NO breakdown product), lipase (pancreas injury), lactate dehydrogenase, and transaminases (liver injury) were measured 16 h after LPS. Although both inhibitors reduced plasma nitrate, they produced contrasting effects on survival and organ injury. L-NAME enhanced liver damage and tended to accelerate the time of death, while L-canavanine significantly reduced mortality and had no deleterious effects in terms of organ damage. These results indicate that nonselective NOS inhibitors are detrimental in endotoxic shock and support the potential usefulness of selective inducible NOS inhibitors in this setting. PMID- 9419180 TI - Susceptibility to leprosy is linked to the human NRAMP1 gene. AB - Leprosy is a debilitating infectious disease of human skin and nerves. Genetic factors of the host play an important role in the manifestation of disease susceptibility. The human NRAMP1 gene is a leprosy susceptibility candidate locus since its murine homologue Nramp1 (formerly Lsh/Ity/Bcg) controls innate resistance to Mycobacterium lepraemurium. In this study, 168 members of 20 multiplex leprosy families were genotyped for NRAMP1 alleles and 4 closely linked polymorphic markers. Highly informative haplotypes overlapping the NRAMP1 gene were constructed, and the haplotype segregation into leprosy-affected offspring was analyzed. It was observed that the segregation of NRAMP1 haplotypes into affected siblings was significantly nonrandom. This finding is consistent with the hypothesis that NRAMP1 itself is a leprosy susceptibility locus. PMID- 9419181 TI - The impact of penicillinase on cefamandole treatment and prophylaxis of experimental endocarditis due to methicillin-resistant Staphylococcus aureus. AB - Beta-lactams active against methicillin-resistant Staphylococcus aureus (MRSA) must resist penicillinase hydrolysis and bind penicillin-binding protein 2A (PBP 2A). Cefamandole might share these properties. When tested against 2 isogenic pairs of MRSA that produced or did not produce penicillinase, MICs of cefamandole (8-32 mg/L) were not affected by penicillinase, and cefamandole had a > or =40 times greater PBP 2A affinity than did methicillin. In rats, constant serum levels of 100 mg/L cefamandole successfully treated experimental endocarditis due to penicillinase-negative isolates but failed against penicillinase-producing organisms. This suggested that penicillinase produced in infected vegetations might hydrolyze the drug. Indeed, cefamandole was slowly degraded by penicillinase in vitro. Moreover, its efficacy was restored by combination with sulbactam in vivo. Cefamandole also uniformly prevented MRSA endocarditis in prophylaxis experiments, a setting in which bacteria were not yet clustered in the vegetations. Thus, while cefamandole treatment was limited by penicillinase, the drug was still successful for prophylaxis of experimental MRSA endocarditis. PMID- 9419182 TI - Listeriosis outbreak associated with the consumption of rillettes in France in 1993. AB - An outbreak of listeriosis involving 38 patients occurred in France between 18 June and 5 October 1993. The epidemic clone was characterized by serovar 4b, phagovar 2671:108:312, and DNA macrorestriction patterns 12 and 13. Thirty-one case-patients were materno-neonatal patients and 7 patients were nonpregnant adults. Preliminary analysis of a case-control study implicated a pork product, rillettes, of a particular brand (odds ratio, 18; 95% confidence interval, 2.2 208) as the vehicle of infection. Rillettes is a ready-to-eat food prepared with ham meat cooked with grease. The implicated lots of rillettes were recalled in mid-August, and the French authorities issued a warning to the general public. Microbiologic analysis of unopened plastic cans of rillettes confirmed the results of the case-control study 3 weeks after the recall. Final analysis showed that the rillettes was the major vehicle of the outbreak but suggested that other brand A meat products could also have been involved. PMID- 9419183 TI - Enhanced control of an outbreak of Mycoplasma pneumoniae pneumonia with azithromycin prophylaxis. AB - There are currently no recommended epidemic-control measures for Mycoplasma pneumoniae pneumonia outbreaks in closed communities. Previous studies have suggested the usefulness of chemoprophylaxis administered to close contacts of case-patients. To evaluate the effectiveness of various epidemic-control measures during an institutional outbreak, an observational study was undertaken during a very large outbreak of M. pneumoniae pneumonia at a facility for developmentally disabled residents (n = 142 cases). Control measures evaluated included no control, standard epidemic-control measures, and targeted azithromycin prophylaxis (500 mg on day 1, 250 mg/day on days 2-5) plus standard epidemic control measures. The combined use of azithromycin prophylaxis and standard epidemic-control measures was associated with a significant reduction in the secondary attack rate. This study suggests that the addition of antibiotic prophylaxis to standard epidemic-control measures can be useful during institutional outbreaks of M. pneumoniae pneumonia. PMID- 9419184 TI - Increase in endocervical CD4 lymphocytes among women with nonulcerative sexually transmitted diseases. AB - To assess associations of nonulcerative sexually transmitted diseases (STDs) with human immunodeficiency virus (HIV)-susceptible leukocytes on female genital mucosa, cervicovaginal specimens from 32 HIV-negative STD clinic patients with gonorrhea, chlamydial infection, or trichomoniasis were compared with specimens from 32 clinic patients without these infections. Twenty-eight patients had single infections (15 gonorrhea, 10 chlamydial infection, 3 trichomoniasis), and 4 had dual infections. A saline vaginal wash and saline suspensions of vaginal wall scrapings, ectocervical scrapings, and endocervical brushings were analyzed by flow cytometry. Specimens from the endocervix had the highest proportions of lymphocytes, monocytes, and Langerhans' cells. The median number of endocervical CD4 lymphocytes/10,000 cells was greater among patients with STDs than among those without (476 vs. 245; P < .001). These data suggest that the endocervix may have a particularly important role in heterosexual HIV transmission and that nonulcerative STDs may facilitate HIV transmission by increasing the presence of CD4 lymphocytes at this site. PMID- 9419185 TI - Recombinant murine granulocyte colony-stimulating factor protects against acute disseminated Candida albicans infection in nonneutropenic mice. AB - The effect of recombinant granulocyte colony-stimulating factor (rG-CSF) on acute disseminated Candida albicans infection in nonneutropenic mice was investigated. Mice treated with a single dose of rG-CSF showed a significantly reduced mortality (28% vs. 90%; P < .001). The outgrowth of C. albicans from the kidneys, spleens, and livers of rG-CSF-treated mice was significantly reduced (log cfu/g of kidney, 5.54 vs. 7.13; P < .001), as were circulating tumor necrosis factor alpha and interleukin-1beta. After rG-CSF, the kidneys showed fewer infectious infiltrates, enhanced granulocyte influx, and almost complete absence of hyphal outgrowth. During peritoneal C. albicans infection, rG-CSF enhanced influx of granulocytes to the site of infection, and exudate granulocytes showed increased oxygen radical production. These results indicate that rG-CSF enhances host resistance to disseminated candidiasis in nonneutropenic mice through activation of granulocytes and their recruitment to the site of infection. PMID- 9419186 TI - Pneumocystis carinii glycoprotein A inhibits surfactant phospholipid secretion by rat alveolar type II cells. AB - Pneumocystis carinii pneumonia (PCP) remains a major cause of morbidity in AIDS. The pathogenesis of PCP is poorly understood, but evidence of surfactant abnormalities is mounting. The role of the major surface glycoprotein of P. carinii, gpA, in producing surfactant abnormalities was investigated. Rat type II pneumocytes were incubated with [3H]choline, purified gpA, and modulators. Lipid was extracted, and [3H]dipalmitoyl phosphatidylcholine (DPPC) secretion was calculated. Contaminating endotoxin had no effect on DPPC secretion. gpA inhibited basal and ATP-stimulated DPPC secretion in a dose- and time-dependent manner. An anti-gpA monoclonal antibody attenuated inhibition of DPPC secretion. Unglycosylated recombinant gpA inhibited secretion, suggesting that functional activity resides in the protein moiety of gpA. These results suggest that gpA is a specific trigger for abnormalities of surfactant lipids in PCP. This is a unique role for a microbial product in disease pathogenesis and a potentially exploitable therapeutic target. PMID- 9419187 TI - Protection against Leishmania major challenge infection in mice vaccinated with live recombinant parasites expressing a cytotoxic gene. AB - A "suicide" system based on thymidine kinase-ganciclovir combination was developed and tested in a Leishmania major experimental model. Susceptible BALB/c mice were infected with L. major expressing the thymidine kinase gene of herpes simplex virus type 1 and treated for 2 consecutive weeks with 7.5 mg/kg/day ganciclovir at different times from the initial infection. Ganciclovir treatment at varying times after infection had different effects on the outcome of disease. A complete inhibition of intracellular parasites was obtained in mice treated 1 or 4 days after infection, whereas ganciclovir administration 2 weeks later resulted in the control of infection only when the drug was provided. Variable levels of protection, from partial to total, against challenge infection with virulent L. major were observed, depending on the timing of ganciclovir treatment. The thymidine kinase-ganciclovir approach represents an excellent experimental model to control Leishmania infection and to evaluate the immunologic response of the host. PMID- 9419188 TI - Recombinant cDNA clones for immunodiagnosis of strongyloidiasis. AB - Because diagnosis of strongyloidiasis by stool examination is unreliable and because of the potential for serious disease in Strongyloides infections, there is need for improved diagnostic aids to facilitate recognition and treatment of this parasitic infection. Serologic testing, when available, requires antigen preparation from infected primates or dogs that can be difficult to maintain. Several recombinant clones from a cDNA library prepared from the infective stage of Strongyloides stercoralis were characterized. Serologic results indicate that the recombinant proteins were equally or more reactive than the larval somatic antigen. No cross-reactivity with recombinant antigen 5a was found with sera from patients with filarial or intestinal nematode infections. Recombinant antigens 5a and 12a detected parasite-specific IgE and IgG4 antibodies in Strongyloides infected patients. Sequence analysis showed these antigens to be rich in proline and charged amino acids. Lack of homology from database searches suggests that the antigens are unique. These recombinant antigens should be useful in diagnostic and epidemiologic studies of strongyloidiasis. PMID- 9419189 TI - Genetic analysis of measles viruses isolated in the United States, 1995-1996. AB - Genetic analysis was conducted on 28 wild type measles viruses isolated from outbreaks or cases in the United States during 1995-1996. These viruses were members of at least 6 distinct genetic groups. However, none of these viruses was related to the group 2 viruses that were associated with the resurgence of measles in the United States between 1989 and 1992 except for a single importation from the Philippines. The sequence data support and extend previous findings showing that transmission of group 2 viruses within the United States was interrupted after 1993. The data also suggest that all measles cases that occurred in the United States in 1995-1996 were the result of importation of virus, even in instances when the source was unknown. Molecular epidemiologic studies can provide a means to measure the success of measles control programs by helping to identify the transmission pathways of the virus. PMID- 9419190 TI - Prior infection with cytomegalovirus is not a major risk factor for angiographically demonstrated coronary artery atherosclerosis. AB - To determine if cytomegalovirus (CMV) infection is a risk factor for primary coronary artery disease (CAD), the association between CMV infection and CAD (>50% blockage in any coronary artery) was investigated in nearly 900 successive nontransplant patients undergoing coronary angiography. By use of logistic regression, older age (P <.001), white race (P <.001), gender (P <.001), hypercholesterolemia (P = .04), and other established cardiovascular risk factors (P = .003) were identified as significantly associated with CAD, but CMV seropositivity (P = .462), the level of IgG antibodies to CMV whole cell antigen (P = .98), or the levels of IgG antibodies to CMV glycoprotein B (P = .67) were not. These data suggest that CMV infection is not a major risk factor for the development of primary CAD in adults. PMID- 9419191 TI - Human herpesvirus 8 detection in nasal secretions and saliva. AB - The presence of human herpesvirus 8 (HHV-8) was determined by polymerase chain reaction (PCR) in nasal secretions and saliva from 14 HHV-8-seropositive persons, including 8 Kaposi's sarcoma patients: 7 were human immunodeficiency virus type 1 infected, 6 of whom were asymptomatic. HHV-8 was detected in one or both body fluids in 8 (57%) of 14 subjects. Parallel PCR testing revealed the concomitant presence of cytomegalovirus, Epstein-Barr virus, and HHV-6 in various combinations in these body fluids. These data indicate frequent shedding of multiple herpesviruses in nasal secretions and saliva, particularly in Kaposi's sarcoma patients. Both body fluids are therefore potential sources HHV-8 by nonsexual transmission. PMID- 9419192 TI - A rapid assay to screen for drug-resistant herpes simplex virus. AB - A rapid assay was developed to screen for herpes simplex virus (HSV) isolates that are resistant to acyclovir and other antiviral agents. The assay is a modified plaque reduction assay (PRA) in which the number of plaques seen in the absence of acyclovir was compared with that seen in the presence of a single cutoff concentration of acyclovir (2 microg/mL). This assay utilizes a cell line that expresses beta-galactosidase only after infection with HSV. Since histochemically stained plaques are easily visualized, small plaques can be easily enumerated. This allows the assay to be performed on dilutions of untitered specimens in the small wells of a 24-well plate and allows the results to be read only 2 days after inoculation of the virus. The assay performed well compared with a standard PRA and should be a valuable tool in identifying drug resistant HSV in a timely manner. PMID- 9419193 TI - Immunogenicity of live attenuated SA14-14-2 Japanese encephalitis vaccine--a comparison of 1- and 3-month immunization schedules. AB - Live attenuated SA14-14-2 Japanese encephalitis (JE) vaccine has been safe and effective in >100 million immunized children, but its current administration schedule of two doses given a year apart does not lend itself to inclusion in established Expanded Program of Immunization (EPI) schedules of childhood immunization. Immune responses to immunization at shorter intervals were compared in middle-school-aged children immunized with two doses separated by 1 month (n = 116) or 2.5 months (n = 115). Two vaccine lots were compared. Seroconversion to the vaccine was observed in 100% of vaccinees immunized in the 1-month schedule and in 94% (lot 2) and 100% (lot 1) of vaccinees immunized in the 2.5-month schedule. Geometric mean titers were almost 2-fold higher with the longer schedule. The routine administration of JE SA14-14-2 vaccine to infants in an EPI schedule should be possible using either interval. PMID- 9419194 TI - Association of CD4 cell depletion and elevated blood and seminal plasma human immunodeficiency virus type 1 (HIV-1) RNA concentrations with genital ulcer disease in HIV-1-infected men in Malawi. AB - CD4 cell counts and blood plasma and seminal plasma human immunodeficiency virus type 1 (HIV-1) concentrations were compared in HIV-1 RNA-seropositive men with urethritis and with or without genital ulcer disease (GUD). GUD was associated with lower CD4 cell counts (median, 258 vs. 348/microL) and increased blood plasma HIV-1 RNA (median, 240 x 10[3] vs. 79.4 x 10[3] copies/mL). Men with nongonococcal urethritis and GUD shed significantly greater quantities of HIV-1 in semen (median, 195 x 10[3] vs. 4.0 x 10[3] copies/mL) than men with nongonococcal urethritis without GUD. These levels decreased approximately 4-fold following antibiotic therapy. The results indicate an association between GUD and increased blood HIV-1 RNA levels. Increased HIV-1 in semen was demonstrated in some men with GUD; such an increase could lead to increased transmission, thus complicating interpretation of the role of the genital ulcer itself in the infectiousness of HIV. Reasons for increased HIV RNA in semen in men with GUD remain to be determined. PMID- 9419195 TI - Horizontal and vertical transmission of human immunodeficiency virus type 1 dual infections caused by viruses of subtypes B and C. AB - This article describes a case of horizontal (heterosexual) and subsequent vertical (mother to infant) transmission of 2 human immunodeficiency viruses type 1 (HIV-1) subtypes. Dual infection in a husband, his wife, and their child was initially detected by use of a restriction fragment length polymorphism assay of the proviral protease in peripheral blood mononuclear cells. The simultaneous presence of highly similar sets of HIV-1 subtypes B and C infecting the 3 family members was confirmed by DNA sequence analysis of pol, gag, and env genes. These data, together with available epidemiologic information, may indicate that the husband's high-risk sexual behavior was the source of dual infections. Because his wife did not report such activities, it was likely that he passed HIV-1 strains to his spouse, who subsequently transmitted them to their child. PMID- 9419196 TI - A placebo-controlled trial of ranitidine in patients with early human immunodeficiency virus infection. AB - Previous uncontrolled reports have suggested that H2-antagonists may possess immunomodulatory activity in human immunodeficiency virus (HIV)-infected patients. Such trials reported improvements in HIV-related symptoms, increased absolute CD4 cell numbers, and improvements in other measures of host immunity. The present trial was a randomized, placebo-controlled, double-blind trial of ranitidine 300 mg (orally twice daily) in subjects with early HIV infection (absolute CD4 cells, 400-700/mm3). Eighty-one subjects entered the trial and 73 completed 16 weeks on study medications. There were no significant differences in the time-weighted average change from baseline between the 2 treatment groups in absolute CD4 cell number, plasma HIV RNA level, or most other surrogate markers of HIV infection. Serum beta2-microglobulin levels were significantly lower in placebo than ranitidine recipients. Ranitidine should not be recommended for the treatment of HIV-infected patients unless it is used for established indications. PMID- 9419197 TI - Residual human immunodeficiency virus type 1 RNA in lymphoid tissue of patients with sustained plasma RNA of <200 copies/mL. AB - Human immunodeficiency virus type 1 (HIV-1) RNA was measured in lymph node (LN) mononuclear cells of 50 patients with sustained plasma RNA of <200 copies/mL with therapy. Six patients had received a combination of three reverse transcriptase inhibitors (RTIs) since primary infection, 11 received this same combination during chronic disease, 21 received a combination of two RTIs plus a protease inhibitor (PI), and 12 received three RTIs plus a PI. The mean overall duration of therapy was 8.9 +/- 0.5 months (range, 5-24), with no significant difference between groups. LN HIV-1 RNA levels varied from undetectable to 1.7 million copies/10(6) cells according to cases. The mean LN HIV-1 RNA level was 2.99 +/- 0.42 log10 copies/10(6) cells in the 17 patients receiving three RTIs compared with 1.93 +/- 0.25 log10 copies/10(6) cells in the 33 patients receiving a PI (t test, P = .02). These data demonstrate that highly active antiretroviral regimens have unequivalent effects on LNs and invite redefinition of suboptimal therapy at this level. PMID- 9419198 TI - Proliferative and cytokine responses of human T lymphocytes isolated from human immunodeficiency virus-infected patients to the major surface glycoprotein of Pneumocystis carinii. AB - The current study examined the proliferative capacity and cytokine secretion pattern of peripheral blood mononuclear cells (PBMC) from human immunodeficiency virus type 1 (HIV-1)-infected patients in response to the major surface glycoprotein (MSG) of Pneumocystis carinii. PBMC from AIDS patients with <200 CD4 cells/mL had significantly less proliferative responses to MSG than did healthy controls. Cytokine analysis indicated that interferon-gamma secreted in response to MSG was also significantly less. There was no significant difference in interleukin-4 levels following incubation with MSG between any of the groups; however, all the HIV-infected persons had slightly elevated levels. When the CDC class C3 patients who had a previous episode of P. carinii pneumonia were compared with those who had not had a previous episode, there was a significant increase in the proliferative response to MSG and in interleukin-4 secretion. CDC class C3 patients who had a previous episode of P. carinii pneumonia showed a predominately Th2 response to MSG. PMID- 9419200 TI - Virulence and colonization-associated properties of Helicobacter pylori isolated from children and adolescents. AB - Helicobacter pylori isolates from 32 children and adolescents were characterized with respect to putative virulence and colonization-associated properties. Only 3 of the subjects had duodenal ulcer. All but 2 of the remaining 29 had various degrees of chronic gastric inflammation. No significant correlation between degree of inflammation and presence of the cag-pathogenicity island, cytotoxin production, vacA alleles associated with cytotoxin expression, and binding ability to the Lewis(b) (Le[b]) oligosaccharide was found. Only 4 isolates expressed the Le(b)-specific adhesin, of which 3 were also cag region-positive. This is in contrast to adults with gastritis or peptic ulcer disease (or both), in whom most of the H. pylori isolates bind Le(b). In an in situ binding assay H. pylori were less able to adhere to gastric surface mucous cells in biopsies taken from children compared with adults, suggesting a lower expression of the Le(b) oligosaccharide in the children. PMID- 9419199 TI - Cytokines in murine lyme carditis: Th1 cytokine expression follows expression of proinflammatory cytokines in a susceptible mouse strain. AB - The cardiac infiltrate seen in murine Lyme carditis is composed predominantly of macrophages, but small numbers of T cells are also present. To identify the cytokines present in cardiac lesions from susceptible mice, semiquantitative polymerase chain reaction was done on cardiac tissue from mice infected with Borrelia burgdorferi. The temporal expression of proinflammatory and T cell derived cytokines was characterized in cardiac tissue at days 0, 3, 7, 14, 21, and 42 after infection with B. burgdorferi. Early in the course of infection, up regulation of the proinflammatory cytokines interleukin-1beta and tumor necrosis factor-alpha was detected. The Th1 cytokine interferon-gamma appeared after the expression of the proinflammatory cytokines and remained elevated throughout the study. Interleukin-4 was not detectable at any time in cardiac lesions. These data are the first to identify cytokines expressed at the lesional level in murine Lyme carditis and to demonstrate a Th1 pattern of cytokine expression in this lesion. PMID- 9419202 TI - Antibody response to the 60-kDa chlamydial heat-shock protein is associated with scarring trachoma. AB - To determine if serum antibody response to the 60-kDa chlamydial heat-shock protein (Chsp60) was associated with scarring trachoma, responses to Chlamydia trachomatis and to Chsp60 from 148 Gambian subjects with trachomatous scarring and from 148 controls without clinical evidence of disease from trachoma-endemic communities were characterized. Chsp60 response was found in 32% of cases and 16% of controls (P < .001). Although C. trachomatis titer was also higher in cases than controls, the prevalence of Chsp60 response between the 2 groups remained significantly different after stratifying for C. trachomatis titer (weighted odds ratio [OR] = 2.1, P = .02). Chsp60 response and C. trachomatis serovar A titer of > or =128 were independently associated with scarring trachoma. The presence of HLA class II allele DRB1*0701 was positively correlated with Chsp60 response (OR = 2.6, P = .02), and DQB1*0301 and DQB1*0501 were negatively associated (OR = 0.42, P < .001; OR = 0.55, P = .46, respectively). PMID- 9419201 TI - Determinants of rifabutin-associated uveitis in patients treated with rifabutin, clarithromycin, and ethambutol for Mycobacterium avium complex bacteremia: a multivariate analysis. Canadian HIV Trials Network Protocol 010 Study Group. AB - Uveitis occurred in a substantial proportion of AIDS patients receiving rifabutin, 600 mg daily, together with clarithromycin and ethambutol for treatment of Mycobacterium avium complex bacteremia. A case-control study was undertaken to examine potential risk factors for developing uveitis. Of eight parameters examined, only baseline body weight predicted the development of uveitis by both univariate and multivariate analyses (P = .001). The incidence of uveitis was 14% in patients weighing >65 kg, 45% in patients between 55 and 65 kg, and 64% in patients <55 kg. Concomitant therapy with fluconazole, a drug known to raise serum rifabutin concentrations, was not associated with an increased incidence of uveitis. The risk of uveitis was markedly reduced when rifabutin was given at 300 mg daily in combination with clarithromycin and ethambutol. PMID- 9419203 TI - Cryptococcal glucuronoxylomannan induces interleukin (IL)-8 production by human microglia but inhibits neutrophil migration toward IL-8. AB - On the basis of the clinical observation that the cerebrospinal fluid (CSF) of patients with cryptococcal meningitis contains high levels of the chemokine interleukin (IL)-8 but few polymorphonuclear leukocytes (PMNL), the production of IL-8 by cultured brain glial cells after stimulation with two serotypes of cryptococcal capsular polysaccharide glucuronoxylomannan (GXM) was studied, followed by an assessment of the effect of GXM on PMNL migration toward IL-8. GXM serotype A but not D was capable of inducing IL-8 production in human fetal microglial cell but not in astrocyte cultures. When added directly to the PMNL, GXM (both serotypes) potently blocked PMNL migration toward IL-8. The mechanism of GXM's inhibitory effect appeared to involve cross-desensitization. These findings suggest that GXM can induce IL-8 production in the brain but that GXM in the systemic circulation inhibits migration of PMNL toward IL-8. PMID- 9419204 TI - Mutations and deletions within the hepatitis B virus core antigen and locations of B cell recognition sites. PMID- 9419205 TI - Nucleotide sequence at position 1081 of the hemagglutinin-neuraminidase gene in the mumps Urabe vaccine strain. PMID- 9419206 TI - Early murine cytomegalovirus (MCMV) infection induces liver natural killer (NK) cell inflammation and protection through macrophage inflammatory protein 1alpha (MIP-1alpha)-dependent pathways. AB - Natural killer (NK) cells mediate defense against early murine cytomegalovirus (MCMV) infections in liver. The chemokine, macrophage inflammatory protein 1alpha (MIP-1alpha), can promote inflammatory responses. Our studies evaluated contributions of NK cells to early MCMV-induced liver inflammation and MIP-1alpha requirements for inflammation and delivery of antiviral defenses. NK cells were shown to be responsible for focal inflammation, and to be induced to migrate at high levels, in MCMV-infected livers. MIP-1alpha gene expression was elevated at coinciding times, and mice deficient in MIP-1alpha function were dramatically inhibited in both inflammatory and protective liver responses. The results precisely define MIP-1alpha-dependent steps required to achieve NK cell inflammation during, and mechanisms promoting defense against, viral infections in tissues. PMID- 9419207 TI - Identification of Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) as the rosetting ligand of the malaria parasite P. falciparum. AB - Severe Plasmodium falciparum malaria is characterized by excessive sequestration of infected and uninfected erythrocytes in the microvasculature of the affected organ. Rosetting, the adhesion of P. falciparum-infected erythrocytes to uninfected erythrocytes is a virulent parasite phenotype associated with the occurrence of severe malaria. Here we report on the identification by single-cell reverse transcriptase PCR and cDNA cloning of the adhesive ligand P. falciparum erythrocyte membrane protein 1 (PfEMP1). Rosetting PfEMP1 contains clusters of glycosaminoglycan-binding motifs. A recombinant fusion protein (Duffy binding like 1-glutathione S transferase; Duffy binding-like-1-GST) was found to adhere directly to normal erythrocytes, disrupt naturally formed rosettes, block rosette reformation, and bind to a heparin-Sepharose matrix. The adhesive interactions could be inhibited with heparan sulfate or enzymes that remove heparan sulfate from the cell surface whereas other enzymes or similar glycosaminoglycans of a like negative charge did not affect the binding. PfEMP1 is suggested to be the rosetting ligand and heparan sulfate, or a heparan sulfate-like molecule, the receptor both for PfEMP1 binding and naturally formed erythrocyte rosettes. PMID- 9419208 TI - Identification of a CD36-related thrombospondin 1-binding domain in HIV-1 envelope glycoprotein gp120: relationship to HIV-1-specific inhibitory factors in human saliva. AB - Human and non-human primate salivas retard the infectivity of HIV-1 in vitro and in vivo. Because thrombospondin 1 (TSP1), a high molecular weight trimeric glycoprotein, is concentrated in saliva and can inhibit the infectivity of diverse pathogens in vitro, we sought to determine the role of TSP1 in suppression of HIV infectivity. Sequence analysis revealed a TSP1 recognition motif, previously defined for the CD36 gene family of cell adhesion receptors, in conserved regions flanking the disulfide-linked cysteine residues of the V3 loop of HIV envelope glycoprotein gp120, important for HIV binding to its high affinity cellular receptor CD4. Using solid-phase in vitro binding assays, we demonstrate direct binding of radiolabeled TSP1 to immobilized recombinant gp120. Based on peptide blocking experiments, the TSP1-gp120 interaction involves CSVTCG sequences in the type 1 properdin-like repeats of TSP1, the known binding site for CD36. TSP1 and fusion proteins derived from CD36-related TSP1-binding domains were able to compete with radiolabeled soluble CD4 binding to immobilized gp120. In parallel, purified TSP1 inhibited HIV-1 infection of peripheral blood mononuclear cells and transformed T and promonocytic cell lines. Levels of TSP1 required for both viral aggregation and direct blockade of HIV-1 infection were physiologic, and affinity depletion of salivary TSP1 abrogated >70% of the inhibitory effect of whole saliva on HIV infectivity. Characterization of TSP1 gp120 binding specificity suggests a mechanism for direct blockade of HIV infectivity that might be exploited to retard HIV transmission that occurs via mucosal routes. PMID- 9419209 TI - The class I antigen-processing pathway for the membrane protein tyrosinase involves translation in the endoplasmic reticulum and processing in the cytosol. AB - Formation of major histocompatibility complex class I-associated peptides from membrane proteins has not been thoroughly investigated. We examined the processing of an HLA-A*0201-associated epitope, YMDGTMSQV, that is derived from the membrane protein tyrosinase by posttranslational conversion of the sequence YMNGTMSQV. Only YMDGTMSQV and not YMNGTMSQV was presented by HLA-A*0201 on cells expressing full-length tyrosinase, although both peptides have similar affinities for HLA-A*0201 and are transported by TAP. In contrast, translation of YMNGTMSQV in the cytosol, as a minigene or a larger fragment of tyrosinase, led to the presentation of the unconverted YMNGTMSQV. This was not due to overexpression leading to saturation of the processing/conversion machinery, since presentation of the converted peptide, YMDGTMSQV, was low or undetectable. Thus, presentation of unconverted peptide was associated with translation in the cytosol, suggesting that processing of the full-length tyrosinase occurs after translation in the endoplasmic reticulum. Nevertheless, presentation of YMDGTMSQV in cells expressing full-length tyrosinase was TAP (transporter associated with antigen processing) and proteasome dependent. After inhibition of proteasome activity, tyrosinase species could be detected in the cytosol. We propose that processing of tyrosinase involves translation in the endoplasmic reticulum, export of full length tyrosinase to the cytosol, and retransport of converted peptides by TAP for association with HLA-A*0201. PMID- 9419210 TI - Antigen is required for the activation of effector activities, whereas interleukin 2 Is required for the maintenance of memory in ovalbumin-specific, CD8+ cytotoxic T lymphocytes. AB - The mechanisms that maintain memory in T cells are not completely understood. We have investigated the role of antigen and interleukin (IL)-2 in the growth and maintenance of CD8+ T cells using a cytolytic T cell line specific for ovalbumin (OVA)257-264 presented by H-2Kb. This line does not secrete IL-4 or IL-2; hence, stimulation with the OVA-transfected EL4 line (E.G7-OVA) does not induce proliferation without addition of exogenous growth factors. Furthermore, this line can be maintained continuously by weekly addition of irradiated, splenic filler cells and IL-2, with or without E.G7-OVA. Although IL-2 induced proliferation of these cytotoxic T lymphocytes (CTLs), production of interferon gamma and tumor necrosis factor alpha required stimulation of the CTL with E. G7 OVA. The kinetics of lymphokine secretion after stimulation by E. G7-OVA were the same whether the CTL had been maintained with or without antigen (Ag). In addition, both CTL lines killed E.G7-OVA target cells within 4 h. Thus, the effector functions of these CTLs were rapidly induced by T cell receptor (TCR) occupancy. CTLs cultured with or without Ag also served as memory T cells when parked for 100 d in unirradiated, syngeneic recipients without OVA. In the absence of OVA, the precursor frequency was identical in spleens of normal and beta2-microglobulin knockout recipients, but significantly less in IL-2 knockout mice. The decline of memory in the absence of IL-2 supports data from other investigators, suggesting that cell cycling is important to the maintenance of CD8+ T cell memory. These data also suggest that stimulation of OVA-specific CTLs by lymphokines seems to be more important to maintaining memory than stimulation of TCRs by cross-reactive peptides complexed to class I molecules. PMID- 9419211 TI - Somatic hypermutation introduces insertions and deletions into immunoglobulin V genes. AB - During a germinal center reaction, random mutations are introduced into immunoglobulin V genes to increase the affinity of antibody molecules and to further diversify the B cell repertoire. Antigen-directed selection of B cell clones that generate high affinity surface Ig results in the affinity maturation of the antibody response. The mutations of Ig genes are typically basepair substitutions, although DNA insertions and deletions have been reported to occur at a low frequency. In this study, we describe five insertion and four deletion events in otherwise somatically mutated VH gene cDNA molecules. Two of these insertions and all four deletions were obtained through the sequencing of 395 cDNA clones (approximately 110,000 nucleotides) from CD38+IgD- germinal center, and CD38-IgD- memory B cell populations from a single human tonsil. No germline genes that could have encoded these six cDNA clones were found after an extensive characterization of the genomic VH4 repertoire of the tonsil donor. These six insertions or deletions and three additional insertion events isolated from other sources occurred as triplets or multiples thereof, leaving the transcripts in frame. Additionally, 8 of 9 of these events occurred in the CDR1 or CDR2, following a pattern consistent with selection, and making it unlikely that these events were artifacts of the experimental system. The lack of similar instances in unmutated IgD+CD38- follicular mantle cDNA clones statistically associates these events to the somatic hypermutation process (P = 0.014). Close scrutiny of the 9 insertion/deletion events reported here, and of 25 additional insertions or deletions collected from the literature, suggest that secondary structural elements in the DNA sequences capable of producing loop intermediates may be a prerequisite in most instances. Furthermore, these events most frequently involve sequence motifs resembling known intrinsic hotspots of somatic hypermutation. These insertion/deletion events are consistent with models of somatic hypermutation involving an unstable polymerase enzyme complex lacking proofreading capabilities, and suggest a downregulation or alteration of DNA repair at the V locus during the hypermutation process. PMID- 9419212 TI - Mutations in the human lambda5/14.1 gene result in B cell deficiency and agammaglobulinemia. AB - B cell precursors transiently express a pre-B cell receptor complex consisting of a rearranged mu heavy chain, a surrogate light chain composed of lambda5/14.1 and VpreB, and the immunoglobulin (Ig)-associated signal transducing chains, Igalpha and Igbeta. Mutations in the mu heavy chain are associated with a complete failure of B cell development in both humans and mice, whereas mutations in murine lambda5 result in a leaky phenotype with detectable humoral responses. In evaluating patients with agammaglobulinemia and markedly reduced numbers of B cells, we identified a boy with mutations on both alleles of the gene for lambda5/14.1. The maternal allele carried a premature stop codon in the first exon of lambda5/14.1 and the paternal allele demonstrated three basepair substitutions in a 33-basepair sequence in exon 3. The three substitutions correspond to the sequence in the lambda5/14. 1 pseudogene 16.1 and result in an amino acid substitution at an invariant proline. When expressed in COS cells, the allele carrying the pseudogene sequence resulted in defective folding and secretion of mutant lambda5/14.1. These findings indicate that expression of the functional lambda5/14.1 is critical for B cell development in the human. PMID- 9419213 TI - Impairment of T and B cell development by treatment with a type I interferon. AB - Type I interferons alpha and beta, naturally produced regulators of cell growth and differentiation, have been shown to inhibit IL-7-induced growth and survival of B cell precursors in vitro. After confirming an inhibitory effect on B lymphopoiesis in an ex vivo assay, we treated newborn mice with an active IFN alpha2/alpha1 hybrid molecule to assess its potential for regulating B and T cell development in vivo. Bone marrow and splenic cellularity was greatly reduced in the IFN-alpha2/alpha1-treated mice, and B lineage cells were reduced by >80%. The bone marrow progenitor population of CD43+B220+HSA- cells was unaffected, but development of the CD19+ pro-B cells and their B lineage progeny was severely impaired. Correspondingly, IL-7-responsive cells in the bone marrow were virtually eliminated by the interferon treatment. Thymus cellularity was also reduced by >80% in the treated mice. Phenotypic analysis of the residual thymocytes indicated that the inhibitory effect was exerted during the pro-T cell stage in differentiation. In IFN-alpha/beta receptor-/- mice, T and B cell development were unaffected by the IFN-alpha2/alpha1 treatment. The data suggest that type I interferons can reversibly inhibit early T and B cell development by opposing the essential IL-7 response. PMID- 9419214 TI - Persistence of peptide-induced CD4+ T cell anergy in vitro. AB - Clonal T cell unresponsiveness, or anergy, has been proposed as a mechanism of peripheral tolerance in vivo, and as a potential means of curbing unwanted T cell responses. In this study, anergy was induced in a T helper cell (Th) clone reactive to hemoglobin (Hb) peptide 64-76 by coculture of the T cells with live antigen-presenting cells (APCs) and 74L, a peptide analog of Hb(64-76) that contains a single amino acid substitution of leucine for glycine at position 74, or with a low concentration of the agonist ligand. The anergic state was characterized by blunted proliferation and interleukin (IL) 2 production upon restimulation with Hb(64-76), and was not the result of impaired TCR/CD3 downmodulation. The addition of exogenous IL-12 transiently restored proliferation of the anergic lines, but removal of IL-12 from culture returned the T cells to their nonproliferative state. Interestingly, persistence of the anergic phenotype was observed despite biweekly restimulation with antigen, APCs, and IL-2. Thus, T cell unresponsiveness induced by a peptide produced a stable, persistent anergic state in a Th0 clone that was not reversible by stimulation with IL-2 or -12. PMID- 9419215 TI - Immunoproteasome assembly: cooperative incorporation of interferon gamma (IFN gamma)-inducible subunits. AB - LMP2, LMP7, and MECL are interferon gamma-inducible catalytic subunits of vertebrate 20S proteasomes, which can replace constitutive catalytic subunits (delta, X, and Z, respectively) during proteasome biogenesis. We demonstrate that MECL requires LMP2 for efficient incorporation into preproteasomes, and preproteasomes containing LMP2 and MECL require LMP7 for efficient maturation. The latter effect depends on the presequence of LMP7, but not on LMP7 catalytic activity. This cooperative mechanism favors the assembly of homogeneous "immunoproteasomes" containing all three inducible subunits, suggesting that these subunits act in concert to enhance proteasomal generation of major histocompatibility complex class I-binding peptides. PMID- 9419216 TI - The CD3-gammadeltaepsilon and CD3-zeta/eta modules are each essential for allelic exclusion at the T cell receptor beta locus but are both dispensable for the initiation of V to (D)J recombination at the T cell receptor-beta, -gamma, and delta loci. AB - The pre-T cell receptor (TCR) associates with CD3-transducing subunits and triggers the selective expansion and maturation of T cell precursors expressing a TCR-beta chain. Recent experiments in pre-Talpha chain-deficient mice have suggested that the pre-TCR may not be required for signaling allelic exclusion at the TCR-beta locus. Using CD3-epsilon- and CD3-zeta/eta-deficient mice harboring a productively rearranged TCR-beta transgene, we showed that the CD3 gammadeltaepsilon and CD3-zeta/eta modules, and by inference the pre-TCR/CD3 complex, are each essential for the establishment of allelic exclusion at the endogenous TCR-beta locus. Furthermore, using mutant mice lacking both the CD3 epsilon and CD3-zeta/eta genes, we established that the CD3 gene products are dispensable for the onset of V to (D)J recombination (V, variable; D, diversity; J, joining) at the TCR-beta, TCR-gamma, and TCR-delta loci. Thus, the CD3 components are differentially involved in the sequential events that make the TCR beta locus first accessible to, and later insulated from, the action of the V(D)J recombinase. PMID- 9419217 TI - Activity and phenotype of natural killer cells in peptide transporter (TAP) deficient patients (type I bare lymphocyte syndrome). AB - In this paper we describe the function and phenotype of natural killer (NK) lymphocytes from HLA class I-deficient patients. These cells are, as has been previously reported, unable to lyse HLA class I- K562 cells, but are able to perform antibody-dependent cellular cytotoxicity (ADCC), although with lower efficiency as compared to NK cells from normal individuals. Transporter associated to antigen processing (TAP)- NK cells proliferate when cultured in the presence of lymphoblastoid B cells (B-LCs) and interleukin 2 and develop a spectrum of cytotoxicity similar to that of activated normal NK cells. Importantly, activation of the TAP- NK cells induces strong cytotoxicity to autologous B-LCs. Analysis of the phenotype of circulating TAP- NK lymphocytes showed them to display a normal diverse repertoire of HLA class I-specific NK receptors. These receptors were expressed at normal levels, apart from the CD94 NKG2A complex, which appeared to be overexpressed. This latter finding could reflect an adaptation to the low expression of HLA class I molecules. Finally, functional analyses indicated that the inhibitory receptors in TAP- individuals can transduce inhibitory signals. Our results suggest that in vivo, the NK cells of TAP- patients could participate in immune defense, at least through ADCC, but upon activation, may be involved in autoimmune processes. PMID- 9419218 TI - Maximal proliferation of cytotoxic T lymphocytes requires reverse signaling through Fas ligand. AB - Fas ligand (FasL/CD95L) is best known for its role in delivering apoptotic signals through its receptor, Fas (APO-1/CD95). In this study, we present evidence that FasL has a second role as a signaling receptor. Alloantigen specific proliferation by multiple FasL- murine CTL lines is depressed compared to that of FasL+ CTL lines. FasL- CTLs kill efficiently on a per recovered cell basis and can achieve wild-type levels of proliferation upon stimulation by optimal doses of anti-CD3, suggesting the lack of a costimulatory signal during antigen stimulation. To test this hypothesis directly, soluble FasIgG, a fusion protein of murine Fas and human IgG1, was added to FasL+ CTLs to demonstrate that blocking cell surface Fas-FasL interactions mimics the depression observed for FasL- CTLs. In addition, plate-bound FasIgG in conjunction with suboptimal anti CD3 stimulation augments proliferative signals in FasL+ but not FasL- CTLs. In contrast to these results with CD8+ T cells, alloantigen-stimulated FasL- CD4+ T cells proliferate vigorously compared to FasL+ cells. These data demonstrate that reverse signaling through FasL is required for CTLs to achieve maximal proliferation and may provide clues to differences in the homeostatic regulation of activated CD4+ and CD8+ T cells during an immune response. PMID- 9419219 TI - Differential expression of chemokine receptors and chemotactic responsiveness of type 1 T helper cells (Th1s) and Th2s. AB - T helper cells type 1 (Th1s) that produce interferon-gamma predominantly mediate cellular immune responses and are involved in the development of chronic inflammatory conditions, whereas Th2s which produce large amounts of IL-4 and IL 5 upregulate IgE production and are prominent in the pathogenesis of allergic diseases. The precise factors determining whether Th1- or Th2-mediated immune responses preferentially occur at a peripheral site of antigen exposure are largely unknown. Chemokines, a superfamily of polypeptide mediators, are a key component of the leukocyte recruitment process. Here we report that among four CXC (CXCR1-4) and five CC (CCR1-5) chemokine receptors analyzed, CXCR3 and CCR5 are preferentially expressed in human Th1s. In contrast, Th2s preferentially express CCR4 and, to a lesser extent, CCR3. In agreement with the differential chemokine receptor expression, Th1s and Th2s selectively migrate in response to the corresponding chemokines. The differential expression of chemokine receptors may dictate, to a large extent, the migration and tissue homing of Th1s and Th2s. It may also determine different susceptibility of Th1s and Th2s to human immunodeficiency virus strains using different fusion coreceptors. PMID- 9419220 TI - Selective inhibition of Ii-dependent antigen presentation by Helicobacter pylori toxin VacA. AB - A major virulence factor in the stomach chronic infection by Helicobacter pylori is a protein toxin (VacA), which alters cell membrane trafficking of late endosomal/prelysosomal compartments. Its role in the chronic infection established by H. pylori is unknown. To test the possibility that VacA alters antigen processing taking place in prelysosomal compartments, we have used the well-established model of antigen processing and presentation consisting of tetanus toxoid-specific human (CD4(+)) T cells stimulated by autologous antigen pulsed Epstein-Barr virus-transformed B cells. We found that VacA interferes with proteolytic processing of tetanus toxin and toxoid and specifically inhibits the Ii-dependent pathway of antigen presentation mediated by newly synthesized major histocompatibility complex (MHC) class II, while leaving unaffected the presentation pathway dependent on recycling MHC class II. The results presented here suggest that VacA may contribute to the persistence of H. pylori by interfering with protective immunity and that this toxin is a new useful tool in the study of the different pathways of antigen presentation. PMID- 9419221 TI - The path from the RNA world. AB - We describe a sequential (step by step) Darwinian model for the evolution of life from the late stages of the RNA world through to the emergence of eukaryotes and prokaryotes. The starting point is our model, derived from current RNA activity, of the RNA world just prior to the advent of genetically-encoded protein synthesis. By focusing on the function of the protoribosome we develop a plausible model for the evolution of a protein-synthesizing ribosome from a high fidelity RNA polymerase that incorporated triplets of oligonucleotides. With the standard assumption that during the evolution of enzymatic activity, catalysis is transferred from RNA --> RNP --> protein, the first proteins in the "breakthrough organism" (the first to have encoded protein synthesis) would be nonspecific chaperone-like proteins rather than catalytic. Moreover, because some RNA molecules that pre-date protein synthesis under this model now occur as introns in some of the very earliest proteins, the model predicts these particular introns are older than the exons surrounding them, the "introns-first" theory. Many features of the model for the genome organization in the final RNA world ribo-organism are more prevalent in the eukaryotic genome and we suggest that the prokaryotic genome organization (a single, circular genome with one center of replication) was derived from a "eukaryotic-like" genome organization (a fragmented linear genome with multiple centers of replication). The steps from the proposed ribo-organism RNA genome --> eukaryotic-like DNA genome --> prokaryotic-like DNA genome are all relatively straightforward, whereas the transition prokaryotic-like genome --> eukaryotic-like genome appears impossible under a Darwinian mechanism of evolution, given the assumption of the transition RNA --> RNP --> protein. A likely molecular mechanism, "plasmid transfer," is available for the origin of prokaryotic-type genomes from an eukaryotic-like architecture. Under this model prokaryotes are considered specialized and derived with reduced dependence on ssRNA biochemistry. A functional explanation is that prokaryote ancestors underwent selection for thermophily (high temperature) and/or for rapid reproduction (r selection) at least once in their history. PMID- 9419222 TI - Relics from the RNA world. AB - An RNA world is widely accepted as a probable stage in the early evolution of life. Two implications are that proteins have gradually replaced RNA as the main biological catalysts and that RNA has not taken on any major de novo catalytic function after the evolution of protein synthesis, that is, there is an essentially irreversible series of steps RNA --> RNP --> protein. This transition, as expected from a consideration of catalytic perfection, is essentially complete for reactions when the substrates are small molecules. Based on these principles we derive criteria for identifying RNAs in modern organisms that are relics from the RNA world and then examine the function and phylogenetic distribution of RNA for such remnants of the RNA world. This allows an estimate of the minimum complexity of the last ribo-organism-the stage just preceding the advent of genetically encoded protein synthesis. Despite the constraints placed on its size by a low fidelity of replication (the Eigen limit), we conclude that the genome of this organism reached a considerable level of complexity that included several RNA-processing steps. It would include a large protoribosome with many smaller RNAs involved in its assembly, pre-tRNAs and tRNA processing, an ability for recombination of RNA, some RNA editing, an ability to copy to the end of each RNA strand, and some transport functions. It is harder to recognize specific metabolic reactions that must have existed but synthetic and bio energetic functions would be necessary. Overall, this requires that such an organism maintained a multiple copy, double-stranded linear RNA genome capable of recombination and splicing. The genome was most likely fragmented, allowing each "chromosome" to be replicated with minimum error, that is, within the Eigen limit. The model as developed serves as an outgroup to root the tree of life and is an alternative to using sequence data for inferring properties of the earliest cells. PMID- 9419223 TI - Synonymous and nonsynonymous substitutions in mammalian genes: intragenic correlations. AB - Previous investigations indicated that synonymous and nonsynonymous substitution rates are correlated in mammalian genes. In the present work, this correlation has been studied at the intragenic level using a dataset of 48 orthologous genes from species belonging to at least four different mammalian orders. The results obtained show that the intragenic variability in synonymous rates is correlated with that of nonsynonymous rates. Moreover, the variation in GC level (and especially of C level) of silent positions along each gene is correlated with the variation in synonymous rate. These results reinforce the previous conclusions that synonymous and nonsynonymous rates as well as GC levels of silent positions are to some extent under common selective constraints. PMID- 9419224 TI - Compositional properties of homologous coding sequences from plants AB - In this work, we investigated (1) the compositional distributions of all available nuclear coding sequences (and of their three codon positions) of six dicots and four Gramineae; this considerably expanded our knowledge about the differences previously seen between these two groups of plants; (2) the compositional correlations of homologous genes from dicots and from Gramineae, as well as from both groups; all correlations were characterized by very good coefficients, with slopes close to unity in the former two cases and very high in the last; (3) the compositional transition that accompanied the emergence of Gramineae from an ancestral monocot; (4) the compositional correlations between exons and introns, which were very good in Gramineae, but only poor to good in dicots; and (5) the compositional profiles of homologous genes from angiosperms, which were characterized by a series of peaks (exons) and valleys (introns) separated by 15-20% GC. The conservative and transitional modes of compositional evolution in plant genes and their general implications are discussed. PMID- 9419225 TI - Did archaeal and bacterial cells arise independently from noncellular precursors? A hypothesis stating that the advent of membrane phospholipid with enantiomeric glycerophosphate backbones caused the separation of the two lines of descent. AB - One of the most remarkable biochemical differences between the members of two domains Archaea and Bacteria is the stereochemistry of the glycerophosphate backbone of phospholipids, which are exclusively opposite. The enzyme responsible to the formation of Archaea-specific glycerophosphate was found to be NAD(P) linked sn-glycerol-1-phosphate (G-1-P) dehydrogenase and it was first purified from Methanobacterium thermoautotrophicum cells and its gene was cloned. This structure gene named egsA (enantiomeric glycerophosphate synthase) consisted of 1,041 bp and coded the enzyme with 347 amino acid residues. The amino acid sequence deduced from the base sequence of the cloned gene (egsA) did not share any sequence similarity except for NAD-binding region with that of NAD(P)-linked sn-glycerol-3-phosphate (G-3-P) dehydrogenase of Escherichia coli which catalyzes the formation of G-3-P backbone of bacterial phospholipids, while the deduced protein sequence of the enzyme revealed some similarity with bacterial glycerol dehydrogenases. Because G-1-P dehydrogenase and G-3-P dehydrogenase would originate from different ancestor enzymes and it would be almost impossible to interchange stereospecificity of the enzymes, it seems likely that the stereostructure of membrane phospholipids of a cell must be maintained from the time of birth of the first cell. We propose here the hypothesis that Archaea and Bacteria were differentiated by the occurrence of cells enclosed by membranes of phospholipids with G-1-P and G-3-P as a backbone, respectively. PMID- 9419226 TI - Organization and nucleotide sequence of the cluster of five histone genes in the polichaete worm Chaetopterus variopedatus: first record of a H1 histone gene in the phylum Annelida. AB - Histone genes were identified and their nucleotide sequences were determined in the polychaete marine worm Chaetopterus variopedatus. The genes are organized in about 390 clusters of 7.3 kbp. Each cluster contains one copy of the five histone genes. The H1 histone gene present in the clusters is the first ever isolated in the phylum Annelida. The cluster has the unique peculiarity that all genes contain both the replication-dependent and the replication-independent 3' mRNA termination signals. Despite the differences in cluster organization and transcription polarity of the individual histone genes between C. variopedatus and Platynereis dumerilii, the other annelid in which histone genes have been studied, phylogenetic analysis of the encoded amino acid sequences clearly groups together those two organisms in a tree in which the other studied worms find closely related positions on the same evolutionary branch. PMID- 9419227 TI - Molecular evolution of the Myb family of transcription factors: evidence for polyphyletic origin. AB - The Myb family of proteins is a group of functionally diverse transcriptional activators found in both plants and animals that is characterized by a conserved DNA-binding domain of approximately 50 amino acids. Phylogenetic analyses of amino acid sequences of this family of proteins portray very disparate evolutionary histories in plants and animals. Animal Myb proteins have diverged from a common ancestor, while plants appear related only within the DNA-binding domain. Results imply a pattern of modular evolution of the Myb proteins centering on the possession of a helix-turn-helix motif. Based on this it is suggested that Myb proteins are a polyphyletic group related only by a "Myb-box" DNA-binding motif. PMID- 9419228 TI - Evolution of substrate specificities in the P-type ATPase superfamily. AB - P-type ATPases make up a large superfamily of ATP-driven pumps involved in the transmembrane transport of charged substrates. We have performed an analysis of conserved core sequences in 159 P-type ATPases. The various ATPases group together in five major branches according to substrate specificity, and not according to the evolutionary relationship of the parental species, indicating that invention of new substrate specificities is accompanied by abrupt changes in the rate of sequence evolution. A hitherto-unrecognized family of P-type ATPases has been identified that is expected to be represented in all the major phyla of eukarya. PMID- 9419229 TI - Evolutionary history of the 11p15 human mucin gene family. AB - The four human mucin genes MUC6, MUC2, MUC5AC, and MUC5B are located at chromosome 11p15.5. It has been demonstrated that the three mucins MUC2, MUC5AC, and MUC5B contain several Cys-subdomains of 108 amino acid residues. In contrast, little information is available concerning MUC6. These Cys-subdomains contain 10 cysteine residues that have a highly conserved position. We present here a coherent probable evolutionary history of this human gene family after comparison of the nucleotide sequences of these Cys-subdomains. The three MUC loci MUC2, MUC5AC, and MUC5B may have evolved from a common ancestral gene by two successive duplications. Moreover, we can postulate that MUC5AC and MUC5B have evolved in a concerted manner, while MUC2 has evolved separately. PMID- 9419230 TI - Toward a better knowledge of the molecular evolution of phosphoenolpyruvate carboxylase by comparison of partial cDNA sequences. AB - To get deeper insight into the evolution of phosphoenolpyruvate carboxylase we have identified PEPC fragments (about 1,100 bp) of another 12 plants species not yet investigated in this context. The selected plants include one Chlorophyta, two Bryophyta, four Pteridophyta, and five Spermatophyta species. The obtained phylogenetic trees on PEPC isoforms are the most complete ones up to now available. Independent of their manner of construction, the resulting dendrograms are very similar and fully consistent with the main topology as it is postulated for the evolution of the higher terrestrial plants. We found a distinct clustering of the PEPC sequences of the prokaryotes, the algae, and the spermatophytes. PEPC isoforms of the archegoniates are located in the phylogenetic trees between the algae and spermatophytes. Our results strengthen the view that the PEPC is a very useful molecular marker with which to visualize phylogenetic trends both on the metabolic and organismic levels. PMID- 9419231 TI - Low divergence in rDNA ITS sequences among five species of Fucus (Phaeophyceae) suggests a very recent radiation. AB - Sequences from the two ribosomal DNA internal transcribed spacers (ITS1 and ITS2) were compared among five species of Fucus. Based on the present taxon sampling, parsimony analysis showed that Fucus serratus is the sister-group of the remaining Fucus species when Ascophyllum nodosum was used as an outgroup. The topology of the tree was (Fucus serratus (F. lutarius (F. vesiculosus (F. spiralis + F. ceranoides)))). The extremely low variation observed suggests a very recent radiation of the genus which supports the view widely accepted that the Fucales are among the most evolutionarily advanced of the brown algae. We further note that sequence differences between Fucus and Ascophyllum were 28%: this does not rule out the utility of ITS sequences within the Fucaceae. The very low number of informative positions allows to demonstrate empirically that distance matrix methods group on the basis of symplesiomorphies. PMID- 9419232 TI - Evaluating evolutionary divergence with microsatellites. AB - We report the use of microsatellites (MS) to track the recent evolution of swine. Allelic frequencies for nine MS loci linked on swine chromosome 6 (SSC6) representing four western and one Chinese swine breeds were used to estimate genetic distances and times of breed divergence. A phylogenetic tree was constructed which partitioned into western and Meishan breed branches. Yorkshire and Hampshire breeds exhibited the most recent divergence with a calculated distance of 391 years. The oldest divergence, of 2,227 years, was between Meishan and Hampshire swine. Estimates of breed divergence are consistent with historical records. Additional analysis suggests that polymorphic MS linked on a single chromosome are sufficient to determine evolutionary relationships within a single species. PMID- 9419233 TI - The parthenogenetic rock lizard lacerta unisexualis: An example of limited genetic polymorphism AB - Protein electrophoresis of Lacerta unisexualis from three populations found that 21 of 36 allozyme loci were homozygous, while 14 expressed fixed heterozygotes and one locus was variable. Three clones were detected at the locus Cat-A. Two individuals represent two rare clones while all others form a common clone. Our favored explanation is the mutation of a preexisting common clone rather than multiple origins. PMID- 9419234 TI - On the RNA world: evidence in favor of an early ribonucleopeptide world. AB - A highly complex RNA world, as is sometimes presented in view of the widespread and diversified use of RNA enzymes, would have encountered many difficulties in passing to a world with catalysis mediated by proteins. These difficulties can be overcome by postulating a very early relationship between the nucleotide and the amino acid components. In particular, after asserting that some characteristics expressed by (nucleotide) coenzymes in catalysis are easier to understand if a close and early relationship between these coenzymes and amino acids is hypothesized, a model is presented for the origin of the enzyme-coenzyme complex. This model is essentially based on an intermediate formed by a tRNA-like molecule covalently linked to a polypeptide. The model attributes the majority of the catalytic role in the ribonucleoprotein world to the latter complex and thus it takes into account the birth of the key intermediate in the origin of protein synthesis-namely, peptidyl-tRNA, which would have otherwise been extremely difficult to select. The predictions of the model are discussed along with its robustness, using the data derived from the study of intermediary metabolism and those from molecular biology. Finally, the appearance of the genetic code in the late phase of the ribonucleopeptide world is discussed. PMID- 9419235 TI - Rapid evolution of a homeodomain: evidence for positive selection. AB - One often-noted feature of homeobox genes is the conservation of the homeodomain among orthologous genes from distantly related species. This sequence conservation is presumed to reflect functional conservation, which indeed has been demonstrated in several cases. We analyzed the evolution of an orphan homeobox gene, Pem, which is expressed preferentially in male and female reproductive tissue. Sequence analysis of 12 species of mice and rats indicated that the Pem gene has evolved at a remarkably high rate. The most rapidly evolving region of the Pem protein is the amino portion of the homeodomain, including the flexible N-terminal arm, helices I and II, and the linker regions between the helices. In contrast, the third helix, which is known to mediate base specific DNA contacts in other homeodomains, is conserved in the Pem protein. Analysis of the ratio of nonsynonymous and synonymous codon substitution rates within the Pem homeodomain suggested that its divergence was driven by adaptive selection. The rate of nonsynonymous substitutions in Pem was higher than that of the sex-determination gene Sry, which also appears to have undergone directional selection over a short evolutionary period. Despite the rapid evolution of the Pem gene, we detected no Pem polymorphisms and observed no variation in the homeobox sequence among closely related Mus species. This suggests that purifying episodes followed phases in which selection pressure drove the rapid divergence of this locus. We propose that transcription factors that function in reproductive events can be subject to rapid adaptive selection. PMID- 9419236 TI - Mhc allelic diversity and modern human origins. AB - Thirty complete coding sequences of human major histocompatibility complex (Mhc) class II DRB alleles, spanning 237 codons, were analyzed for phylogenetic information using distance, parsimony, and likelihood approaches. Allelic genealogies derived from different parts of the coding sequence (exon 2, the 5' and 3' ends of exon 2, respectively, and exons 3-6) were compared. Contrary to prior assertions, a rigorous analysis of allelic genealogies in this gene family cannot be used to justify the claim that the lineage leading to modern humans contained on average at least 100,000 individuals. Phylogenetic inferences based upon the exon 2 region of the DRB loci are complicated by selection and recombination, so this part of the gene does not provide a complete and accurate view of allelic relationships. Attempts to reconstruct human history from genetic data must use realistic models which consider the complicating factors of nonequilibrium populations, recombination, and different patterns of selection. PMID- 9419237 TI - The evolution of MHC diversity by segmental duplication and transposition of retroelements. AB - Sequence analysis of a 237 kb genomic fragment from the central region of the MHC has revealed that the HLA-B and HLA-C genes are contained within duplicated segments peri-B (53 kb) and peri-C (48 kb), respectively, and separated by an intervening sequence (IF) of 30 kb. The peri-B and peri-C segments share at least 90% sequence homology except when interrupted by insertions/deletions including Alu, L1, an endogenous retrovirus, and pseudogenes. The sequences of peri-B, IF, and peri-C were searched for the presence of Alu elements to use as markers of evolution, chromosomal rearrangements, and polymorphism. Of 29 Alu elements, 14 were identified in peri-B, 11 in peri-C, and 4 in IF. The Alu elements in peri-B and peri-C clustered phylogenetically into two clades which were classified as "preduplication" and "postduplication" clades. Four Alu J elements that are shared by peri-B and peri-C and are flanked by homologous sequences in their paralogous locations, respectively, clustered into a "preduplication" clade. By contrast, the majority of Alu elements, which are unique to either peri-B or peri C, clustered into a postduplication clade together with the Alu consensus subfamily members ranging from platyrrhine-specific (Spqxcg) to catarrhine specific Alu sequences (Y). The insertion of platyrrhine-specific Alu elements in postduplication locations of peri-B and peri-C implies that these two segments are the products of a duplication which occurred in primates prior to the divergence of the New World primate from the human lineage (35-44 mya). Examination of the paralogous Alu integration sites revealed that 9 of 14 postduplication Alu sequences have produced microsatellites of different length and sequence within the Alu 3'-poly A tail. The present analysis supports the hypothesis that HLA-B and HLA-C genes are products of an extended segmental duplication between 44 and 81 million years ago (mya), and that subsequent diversification of both genomic segments occurred because of the mobility and mutation of retroelements such as Alu repeats. PMID- 9419238 TI - Unexpected conservation of the X-linked color vision gene in nocturnal prosimians: evidence from two bush babies. AB - Bush babies have had a long history of nocturnal life and it would be interesting to know whether their color vision genes have become degenerate. Therefore, we used PCR techniques to sequence the X-linked pigment gene of two of these nocturnal prosimians: Galago senegalensis and Otolemur garnettii. Southern hybridization of genomic DNA of G. senegalensis showed a single X-linked pigment gene. Interestingly, the deduced pigment sequences of the two bush babies are identical. By comparing the X-linked pigments of bush baby, human, squirrel monkey, and marmoset, 38 variable positions were identified. At those positions that may cause a spectral shift, the bush baby pigment has identical or biochemically similar residues to those of the marmoset cone pigment with a spectral peak of 543 nm. This result is consistent with the estimate of 544-545 nm for the spectral peak of the X-linked pigment of Otolemur crassicaudatus, which is closely related to Otolemur garnettii. The neighbor-joining tree of mammalian X-linked pigments showed a significantly shorter branch in the bush baby lineage than in other primate lineages. A relative rate test showed that the nonsynonymous substitution rate of the bush baby X-linked pigment gene is about three times slower than that of the human red pigment gene, though the synonymous substitution rates of the two genes are similar. The slower nonsynonymous rate in the bush baby lineage suggests that the bush baby X-linked pigment gene is under functional constraints, in spite of its nocturnal life. Two radical changes at positions in the intradiskal surface next to the sixth transmembrane domain were observed in the X-linked cone pigment of bush babies but not in other primates. They are changes from Ala to Ser and from Asn to His, which are similar in function to the corresponding residues in rhodopsins. These two changes may be of importance for dim light sensitivity, which is consistent with our proposal that the evolution of the bush baby X-linked pigment gene is under selective pressure. In addition, the 2.5% divergence in introns 2 and 5 of the X-linked pigment gene between the two bush babies supports their classification into two separate genera. PMID- 9419239 TI - Evolutionary relationships among the eukaryotic crown taxa taking into account site-to-site rate variation in 18S rRNA. AB - In this study we constructed a bootstrapped distance tree of 500 small subunit ribosomal RNA sequences from organisms belonging to the so-called crown of eukaryote evolution. Taking into account the substitution rate of the individual nucleotides of the rRNA sequence alignment, our results suggest that (1) animals, true fungi, and choanoflagellates share a common origin: The branch joining these taxa is highly supported by bootstrap analysis (bootstrap support [BS] > 90%), (2) stramenopiles and alveolates are sister groups (BS = 75%), (3) within the alveolates, dinoflagellates and apicomplexans share a common ancestor BS > 95%), while in turn they both share a common origin with the ciliates (BS > 80%), and (4) within the stramenopiles, heterokont algae, hyphochytriomycetes, and oomycetes form a monophyletic grouping well supported by bootstrap analysis (BS > 85%), preceded by the well-supported successive divergence of labyrinthulomycetes and bicosoecids. On the other hand, many evolutionary relationships between crown taxa are still obscure on the basis of 18S rRNA. The branching order between the animal-fungal-choanoflagellates clade and the chlorobionts, the alveolates and stramenopiles, red algae, and several smaller groups of organisms remains largely unresolved.When among-site rate variation is not considered, the inferred tree topologies are inferior to those where the substitution rate spectrum for the 18S rRNA is taken into account. This is primarily indicated by the erroneous branching of fast-evolving sequences. Moreover, when different substitution rates among sites are not considered, the animals no longer appear as a monophyletic grouping in most distance trees. PMID- 9419240 TI - Characterization of mitochondrial small-subunit ribosomal RNAs from holoparasitic plants. AB - Mitochondrial small-subunit (19S) rDNA sequences were obtained from 10 angiosperms to further characterize sequence divergence levels and structural variation in this molecule. These sequences were derived from seven holoparasitic (nonphotosynthetic) angiosperms as well as three photosynthetic plants. 19S rRNA is composed of a conservative core region (ca. 1450 nucleotides) as well as two variable regions (V1 and V7). In pairwise comparisons of photosynthetic angiosperms to Glycine, the core 19S rDNA sequences differed by less than 1.4%, thus supporting the observation that variation in mitochondrial rDNA is 3-4 times lower than seen in protein coding and rDNA genes of other subcellular organelles. Sequences representing four distinct lineages of nonasterid holoparasites showed significantly increased numbers of substitutions in their core 19S rDNA sequences (2.3-7.6%), thus paralleling previous findings that showed accelerated rates in nuclear (18S) and plastid (16S) rDNA from the same plants. Relative rate tests confirmed the accelerated nucleotide substitution rates in the holoparasites whereas rates in nonparasitic plants were not significantly increased. Among comparisons of both parasitic and nonparasitic plants, transversions outnumbered transitions, in many cases more than two to one. The core 19S rRNA is conserved in sequence and structure among all nonparasitic angiosperms whereas 19S rRNA from members of holoparasitic Balanophoraceae have unique extensions to the V5 and V6 variable domains. Substitution and insertion/deletion mutations characterized the V1 and V7 regions of the nonasterid holoparasites. The V7 sequence of one holoparasite (Scybalium) contained repeat motifs. The cause of substitution rate increases in the holoparasites does not appear to be a result of RNA editing, hence the underlying molecular mechanism remains to be fully documented. PMID- 9419241 TI - The molecular evolution of the vertebrate trypsinogens. AB - We expand the already large number of known trypsinogen nucleotide and amino acid sequences by presenting additional trypsinogen sequences from the tunicate (Boltenia villosa), the lamprey (Petromyzon marinus), the pufferfish (Fugu rubripes), and the frog (Xenopus laevis). The current array of known trypsinogen sequences now spans the entire vertebrate phylogeny. Phylogenetic analysis is made difficult by the presence of multiple isozymes within species and rates of evolution that vary highly between both species and isozymes. We nevertheless present a Fitch-Margoliash phylogeny constructed from pairwise distances. We employ this phylogeny as a vehicle for speculation on the evolution of the trypsinogen gene family as well as the general modes of evolution of multigene families. Unique attributes of the lamprey and tunicate trypsinogens are noted. PMID- 9419242 TI - Deuterostomic actin genes and the definition of the chordates: cDNA cloning and gene organization for cephalochordates and hemichordates. AB - The evolutionary relationship of muscle and nonmuscle actin isoforms in deuterostomia was studied by the isolation and characterization of two actin genes from the cephalochordate Branchiostoma lanceolatum and two from the hemichordate Saccoglossus kowalevskii The Branchiostoma genes specify a muscle and a nonmuscle actin type, respectively. Together with earlier results on muscle actins from vertebrates and urochordates, a N-terminal sequence signature is defined for chordate muscle actins. These diagnostic amino acid residues separate the chordates from the echinoderms and other metazoa. Although the two Saccoglossus actins characterized so far lack the diagnostic residues, in line with the presumptive phylogenetic position of hemichordates outside the chordates, a definitive conclusion can only be expected once the full complement of actin genes of Saccoglossus is established. Comparison of the intron patterns of the various deuterostomic actin genes shows that intron 330-3, which is present in all vertebrate genes, is conspicuously absent from nonvertebrate genes. The possible origin of this intron is discussed. PMID- 9419243 TI - On the origin of protein synthesis factors: a gene duplication/fusion model. AB - Sequence similarity has given rise to the proposal that IF-2, EF-G, and EF-Tu are related through a common ancestor. We evaluate this proposition and whether the relationship can be extended to other factors of protein synthesis. Analysis of amino acid sequence similarity gives statistical support for an evolutionary affiliation among IF-1, IF-2, IF-3, EF-Tu, EF-Ts, and EF-G and suggests further that this association is a result of gene duplication/fusion events. In support of this mechanism, the three-dimensional structures of IF-3, EF-Tu, and EF-G display a predictable domain structure and overall conformational similarity. The model that we propose consists of three consecutives duplication/fusion events which would have taken place before the divergence of the three superkingdoms: eubacteria, archaea, and eukaryotes. The root of this protein superfamily tree would be an ancestor of the modern IF-1 gene sequence. The repeated fundamental motif of this protein superfamily is a small RNA binding domain composed of two alpha-helices packed along side of an antiparallel beta-sheet. PMID- 9419244 TI - On the evolution of the single-subunit RNA polymerases. AB - Many eukaryotic nuclear genomes as well as mitochondrial plasmids contain genes displaying evident sequence similarity to those encoding the single-subunit RNA polymerase (ssRNAP) of bacteriophage T7 and its relatives. We have collected and aligned these ssRNAP sequences and have constructed unrooted phylogenetic trees that demonstrate the separation of ssRNAPs into three well-defined and nonoverlapping clusters (phage-encoded, nucleus-encoded, and plasmid-encoded). Our analyses indicate that these three subfamiles of T7-like RNAPs shared a common ancestor; however, the order in which the groups diverged cannot be inferred from available data. On the basis of structural similarities and mutational data, we suggest that the ancestral ssRNAP gene may have arisen via duplication and divergence of a DNA polymerase or reverse transcriptase gene. Considering the current phylogenetic distribution of ssRNAP sequences, we further suggest that the origin of the ancestral ssRNAP gene closely paralleled in time the introduction of mitochondria into eukaryotic cells through a eubacterial endosymbiosis. PMID- 9419245 TI - The origin of chlorarachniophyte plastids, as inferred from phylogenetic comparisons of amino acid sequences of EF-Tu. AB - A molecular phylogenetic analysis of elongation factor Tu (EF-Tu) proteins from plastids was performed in an attempt to identify the origin of chlorarachniophyte plastids, which are considered to have evolved from the endosymbiont of a photosynthetic eukaryote. Partial sequences of the genes for plastid EF-Tu proteins (1,080-1,089 bp) were determined for three algae that contain chlorophyll b, namely, Gymnochlora stellata (Chlorarachniophyceae), Bryopsis maxima (Ulvophyceae), and Pyramimonas disomata (Prasinophyceae). The deduced amino acid sequences were used to construct phylogenetic trees of the plastid and bacterial EF-Tu proteins by the maximum likelihood, the maximum parsimony, and the neighbor joining methods. The trees obtained in the present analysis suggest that all plastids that contain chlorophyll b are monophyletic and that the chlorarachniophyte plastids are closely related to those of the Ulvophyceae. The phylogenetic trees also suggest that euglenophyte plastids are closely related to prasinophycean plastids. The results indicate that the chlorarachniophyte plastids evolved from a green algal endosymbiont that was closely related to the Ulvophyceae and that at least two secondary endosymbiotic events have occurred in the lineage of algae with plastids that contain chlorophyll b. PMID- 9419246 TI - Analysis of the cluster of ribosomal protein genes in the plastid genome of a unicellular red alga Cyanidioschyzon merolae: translocation of the str cluster as an early event in the rhodophyte-chromophyte lineage of plastid evolution. AB - The nucleotide sequence of a cluster of ribosomal protein genes in the plastid genome of a unicellular red alga, Cyanidioschyzon merolae, which has been supposed to be the most primitive alga, was determined. The phylogenetic tree inferred from the amino acid sequence of ribosomal proteins of two rhodophytes, a chromophyte, a glaucophyte, two chlorophytes (land plants), a cyanobacterium, and three eubacteria suggested a close relationship between the cyanobacterium Synechocystis PCC6803 and the plastids of various species in the kingdom Plantae, which is consistent with the hypothesis of the endosymbiotic origin of plastids. In this tree, the two species of rhodophytes were grouped with the chromophyte, and the glaucophyte was grouped with the chlorophytes. Analysis of the organization of the genes encoding the ribosomal proteins suggested that the translocation of the str cluster occurred early in the lineage of rhodophytes and chromophytes after these groups had been separated from chlorophytes and glaucophytes. PMID- 9419247 TI - Estimation of reversible substitution matrices from multiple pairs of sequences. AB - We present a method for estimating the most general reversible substitution matrix corresponding to a given collection of pairwise aligned DNA sequences. This matrix can then be used to calculate evolutionary distances between pairs of sequences in the collection. If only two sequences are considered, our method is equivalent to that of Lanave et al. (1984). The main novelty of our approach is in combining data from different sequence pairs. We describe a weighting method for pairs of taxa related by a known tree that results in uniform weights for all branches. Our method for estimating the rate matrix results in fast execution times, even on large data sets, and does not require knowledge of the phylogenetic relationships among sequences. In a test case on a primate pseudogene, the matrix we arrived at resembles one obtained using maximum likelihood, and the resulting distance measure is shown to have better linearity than is obtained in a less general model. PMID- 9419248 TI - Beyond the wobble: the rule of conjugates. AB - A reexamination of the genetic code suggests a rule of conjugates which captures the observed quartet degeneracies without exception. Adenine is the conjugate of cytosine and uracil is the conjugate of guanine. Further analysis reveals that the rule of conjugates is a macrolevel manifestation of the molecular-level hydrogen-bonding and base-stacking interactions at the decoding site. This new perspective is of significance to evolutionary discussions of nucleic acid bases, genetic code, and interactions involving RNAs. PMID- 9419249 TI - Bacterial sigma 70 transcription factor DNA-binding domains in the archaeon Methanococcus jannaschii. PMID- 9419250 TI - ThiD-TenA: a gene pair fusion in eukaryotes. AB - Computational analysis of the hypothetical open reading frame MJ0236 from Methanococcus jannaschii reveals its membership to a family of bacterial and eukaryotic proteins, predicted to be the HMP-P kinases involved in thiamin biosyntheis (ThiD). The eukaryotic members of this family contain a C-terminal extension similar to a bacterial transcriptional activator (TenA), thus pointing to a fusion event that took place during cellular evolution. The C-terminal domain is absent from M. jannaschii. The significance of this observation is two fold: first, this is a case where a fusion protein contains two domains with an unusual phylogenetic distribution, and second, the TenA domain is a rare case of a gene family involved in transcription present both in bacteria and eukaryotes. PMID- 9419251 TI - Rodent monophyly: pitfalls of molecular phylogenies. PMID- 9419252 TI - Effects of a lipophilic environmental pollutant (DDT) on the phospholipid and fatty acid contents of Bacillus stearothermophilus. AB - Cultures of Bacillus stearothermophilus grown in a complex medium containing 1 microM DDT, exhibited longer lag adapting periods, decreased specific growth rates, and lower growth yield as compared to control cultures. The membrane lipid composition from cells grown in the presence of the insecticide was significantly different from that of control cells. The effects of DDT (2,2-bis(p-chlorophenyl) 1, 1,1-trichloroethane) on growth and lipid composition of bacterial cells were also determined in cultures grown in a medium supplemented with Ca2+ (membrane stabilizer) to further clarify the influence of growth conditions on bacterial responses to the toxicant. The main membrane-lipid changes induced by DDT relate to a very significant increase (74%) of the relative concentration of a phosphoglycolipid, an increase of the phosphatidylethanolamine content, with a parallel decrease of phosphatidylglycerol and an unidentified phospholipid X0. The changes of the phospholipid acyl chains relate to an increase of straight chains and a parallel decrease of branched chains. The effects of DDT-induced lipid composition alterations on membrane physical properties were monitored by fluorescence polarization studies with bacterial polar lipid dispersions. Changes in the membrane lipids upon growing the bacteria in a DDT-containing medium promoted, as expected, more ordered membranes with a shift of the phase transition temperature to higher values. Data are interpreted in the frame of an adaptation mechanism to counteract the membrane perturbation resulting from the accumulation of the insecticide molecules in the lipid bilayer. PMID- 9419253 TI - Comparative fate of 1,1-diphenylethylene (DPE), 1,1-dichloro-2,2-bis(4 Chlorophenyl)-ethylene (DDE), and pentachlorophenol (PCP) under alternating aerobic and anaerobic conditions. AB - Bacterial degradation of 1,1-dichloro-2,2-bis-(4-chlorophenyl)-ethylene (DDE) and its dehalogenated derivative 1,1-diphenylethylene (DPE) has not yet been shown and may require culture adaptation and special culture conditions. We compared the degradability of DPE, DDE, and pentachlorophenol (PCP) in aerobic/anaerobic sequenced batch reactor systems. Reactors operated under aerobic/methanogenic and aerobic/denitrifying conditions were inoculated with bacterial consortia from anaerobic granular sludge, long-term PCP- and DDE-contaminated soil, and pulp and paper waste pond sediment. The culture was gradually acclimatized to low concentrations of DPE, DDE, and PCP in defined minimal growth media with benzoate, phenol, ethanol, and formate as primary carbon sources. DDE remained refractory for 105 days, whereas DPE and PCP were degraded. This suggests that DDE is extremely recalcitrant to degradation by aromatic organochlorine-degrading bacteria from long-term polluted soils and sediments. The results confirm that the chlorination of DDE is a major biodegradation barrier for adapted bacteria under aerobic and anaerobic conditions. PMID- 9419254 TI - Concentrations of chlorinated organic compounds in biota and bed sediment in streams of the San Joaquin Valley, California. AB - Samples of resident biota and bed sediments were collected in 1992 from 18 sites on or near the floor of the San Joaquin Valley, California, for analysis of 33 organochlorine compounds. The sites were divided into five groups on the basis of physiographic region and land use. Ten compounds were detected in tissue, and 15 compounds were detected in bed sediment. The most frequently detected compound in both media was p,p'-DDE. Concentrations of SigmaDDT (sum of o,p'- and p, p' forms of DDD, DDE, and DDT) were statistically different among groups of sites for both tissue and sediment (Kruskal-Wallis, p < 0.05). Concentrations in both media were highest in streams draining the west side of the valley. Concentrations of SigmaDDT in tissue were significantly correlated with specific conductance, pH, and total alkalinity (p < 0.05), which are indicators of the proportion of irrigation return flows in stream discharge. Concentrations in sediment on a dry weight basis were not correlated with these water-quality parameters, but total organic carbon (TOC) normalized concentrations were significantly correlated with specific conductance and pH (p < 0.05). Regressions of the concentration of SigmaDDT in tissue, as a function of SigmaDDT in bed sediment, were significant and explained up to 76% of the variance in the data. The concentration of SigmaDDT in sediment may be related to mechanisms of soil transport to surface water with bioavailability of compounds related to the concentration of TOC in sediment. The results of this study did not indicate any clear advantage to using either bed sediment or tissues in studies of organochlorine chemicals in the environment. Some guidelines for protection of fish and wildlife were exceeded. Concentrations of organochlorine chemicals in biota, and perhaps sediment, have declined from concentrations measured in the 1970s and 1980s, but remain high compared to other regions of the United States. PMID- 9419255 TI - Concentration and transport of chlordane and nonachlor associated with suspended sediment in the Mississippi River, May 1988 to June 1990. AB - Technical chlordane, a formerly widely used organochlorine pesticide, has become widespread in the environment. The distribution of technical chlordane in riverine environments may be due in part to resuspension and aqueous transport of contaminated bed sediment. To test this hypothesis, the Mississippi River was sampled for suspended sediment five times over a two-year period, at up to 17 sites from St. Louis to below New Orleans, including major tributaries. The ratio of chlordane to nonachlor concentrations averaged 3.6 during May-June 1988 for the Mississippi River below its confluence with the Ohio River. During March April 1989, the ratio was 0.6, suggesting weathered technical chlordane contributions to the suspended sediment. During June 1989, the ratio averaged 1.1, indicating some input of less weathered technical chlordane. During February March and May-June 1990, the ratios again shifted, from 0.8 to 1.3. This shifting ratio is likely due to resuspension of weathered technical chlordane associated with bed sediment during spring runoff. Annual transport by suspended sediment from the Mississippi River to the Gulf of Mexico was estimated to be 110 kg of chlordane and 100 kg of nonachlor. PMID- 9419256 TI - Organochlorine pesticides and polychlorinated biphenyls (PCBs) in sediments and biota from four US Arctic lakes. AB - Organochlorine (OC) concentrations in surface sediment, snails (Lymnea sp.), and two freshwater fish species (grayling, Thymallus arcticus; and lake trout, Salvelinus namaycush) from four lakes in the US Arctic were determined. In surface sediment, chlorinated benzenes (including hexachlorobenzene, HCB), and p,p'-DDT were the primary analytes detected (max = 0.7 ng/g dry wt), while individual polychlorinated biphenyl (PCB) congeners were always below 0.1 ng/g. A wider range of compounds and higher concentrations were found in lake trout, the top predatory fish species in the same lakes. The concentration ranges for hexachlorocyclohexanes (HCHs), chlordane-related compounds (CHLORs), DDTs, and PCBs in lake trout and grayling were similar to those reported for other arctic freshwater fish (1-100 ng/g wet wt), but one to two orders of magnitude lower than Great Lakes salmonids. Nitrogen isotope analysis confirmed that differences in OC concentrations between grayling and lake trout are explained partly by differences in food web position. PMID- 9419257 TI - Comparative bioaccumulation of chlorinated hydrocarbons from sediment by two infaunal invertebrates. AB - Bioaccumulation of chlorinated hydrocarbons (CHs) from field-contaminated sediments by two infaunal invertebrates, Rhepoxynius abronius (a non-deposit feeding amphipod) and Armandia brevis (a nonselective, deposit-feeding polychaete), was examined and species responses were compared. Sediments were selected over a large geographical area of the Hudson-Raritan estuary to assess the potential for bioaccumulation from a typical urban estuary. Unlike polycyclic aromatic hydrocarbons (PAHs) from these sediments, concentrations of CHs in interstitial water (IW) indicated that partition coefficients (Koc) were generally as expected, especially when based on predicted, nonsorbed, interstitial water CH concentrations (IWfree). Correlations between amphipod and polychaete tissue residues revealed that these species were responding similarly to a gradient of CH concentrations in sediment. While tissue residues and BAFloc (lipid/organic carbon normalized bioaccumulation factor) values for the trichlorobiphenyls were similar for both species, accumulation in the polychaete was three to 10 times higher for the more hydrophobic PCBs, which was attributed to differences in the route of exposure. A negative correlation between the bioaccumulation factor (BAF) and total organic carbon (TOC) was found for both species, which was expected according to equilibrium partitioning theory. Because it was assumed that the amphipod was not feeding in these tests and the polychaete was ingesting sediment, comparison of their tissue residues and bioaccumulation factors was useful for highlighting the importance of sediment ingestion, especially for short-term, nonequilibrium exposures. These results may also help elucidate the limitations associated with assessing bioaccumulation and the resultant toxic response in standard 10-day toxicity tests with similar invertebrates. PMID- 9419258 TI - Population growth kinetics of Tetrahymena pyriformis exposed to selected nonpolar narcotics. AB - This study describes effects of selected nonpolar narcotics of varying hydrophobicity (quantified by the 1-octanol-water partition coefficient, log Kow) and molecular structure on the population growth kinetics of the freshwater ciliate Tetrahymena pyriformis. The response of Tetrahymena exposed to different nonpolar narcotics varied from a change in generation time to a change in lag phase with similar generation time compared to control. Two narcotics with high (>3.00), intermediate (>0.00 and <3.00), and low log Kow (<0. 00) values were tested. Growth of Tetrahymena inhibited up to 85% by the high log Kow toxicants (2-decanone and butylbenzene) grew with similar rates as the control, but exhibited increased lag time, suggesting that the protozoan became acclimated to toxicant stress. Results from growth of Tetrahymena in the low log Kow toxicants (ethanol and acetone) indicate an increased generation time with increasing concentration. Cells inhibited by the intermediate log Kow chemicals, 1-pentanol and anisole, exhibited a response that was a combination of the previously mentioned two contrary responses. Cells inhibited <35% with 1-pentanol and <50% with anisole grew with similar generation times as control flasks, whereas in cells inhibited >35% or >50%, respectively, the doubling times were longer than control growth. PMID- 9419259 TI - Evaluation of three larval instars of the midge Chironomus petiolatus as bioassay tools using a computationally intensive statistical algorithm. AB - Sensitivity to toxicants is a major criterion for selecting organisms for bioassay testing. If a sensitive species is also abundant and occupies a role as prey for many other species within a community, then the species become a valuable tool in environmental monitoring. These features apply to larval midge Chironomus petiolatus in freshwater environments of central Chile. The youngest larval instar is the most sensitive and presents the additional feature of lower survival within control arenas, making it more difficult to discern toxicant related mortality from background mortality. In this work, we perform acute bioassays with the three larval stages of C. petiolatus and K2Cr2O7 as reference toxicant, with the goal of selecting a particular instar as the best bioassay tool using two criteria: sensitivity and background mortality. Sensitivity is evaluated through Monte Carlo estimation of LC50 and background mortality through bootstrap resampling, and a final Bioassay Performance Index as the product of LC50 and background mortality. For this task we developed a new computationally intensive statistical algorithm. Results show that the best bioassay tool is not the youngest and most sensitive instar but an intermediate one. PMID- 9419260 TI - Effects of sublethal copper exposure on copper accumulation, food consumption, growth, energy stores, and nucleic acid content in common carp. AB - Juvenile common carp were exposed for 28 days to three different sublethal copper concentrations (0.20 microM, 0.55 microM, and 0.80 microM). Food consumption was monitored on a daily basis during the exposure period, while growth, copper accumulation, energy stores, and nucleic acid contents were assessed weekly. Copper exposure to 0. 80 microM affected both growth and feeding behavior in common carp. At 0.55 microM, growth was affected despite normal food consumption. Even at the lowest copper concentration (0.20 microM), metabolic demand for the fish increased, challenging the carp with an increased demand for food. Copper accumulation mainly occurred in the liver, reaching an equilibrium between uptake and excretion after 1 month of exposure. Substantial biochemical changes were observed at the two highest copper exposure concentrations, but the correlation between growth rate and RNA:DNA ratio was poor considering the substantial differences in growth rate. The use of the RNA:DNA ratio as a sensitive biomarker is questioned. PMID- 9419261 TI - Mercury distribution in sediments and bioaccumulation by fish in two oregon reservoirs: point-source and nonpoint-source impacted systems. AB - Mercury pollution was compared in two Oregon reservoirs of similar size and age, located within the same ecoregion. Cottage Grove Reservoir was distinguished by a history of mercury mining and processing within its watershed, while Dorena Reservoir was not. Mercury concentrations in sediments of the reservoirs, tributary streams, and three species of fish were measured. Sediment mercury concentrations in the main tributary of Cottage Grove Reservoir, which drains the subbasin where past mercury mining occurred, was tenfold higher than mercury in sediments from other reservoir tributaries. There were no significant differences between sediment mercury concentrations in the tributaries of Dorena Reservoir. The average mercury concentration in the basin sediment of Cottage Grove Reservoir (0.67 +/- 0.05 microg/g dry wt) was higher than for Dorena Reservoir (0.12 +/- 0.01 microg/g dry wt). At Cottage Grove Reservoir, maximum mercury concentrations were near or exceeded 1 microg/g wet wt for largemouth bass (Micropterus salmonides) and bluegill (Lepomis macrochirus) epaxial muscle. Muscle mercury concentrations in largemouth bass and crappie (Pomoxis nigromaculatus) from Cottage Grove Reservoir were significantly higher than from the same species from Dorena Reservoir. Numbers of bluegill of the same age available from both reservoirs were too small for statistical comparisons. Mercury concentrations in largemouth bass muscle fluctuated annually in both reservoirs. Fish ages were consistently positively correlated with muscle mercury concentrations in only the point-source-impacted reservoir. These results indicated that a point source, Black Butte Mine, contributed amounts of mercury greatly in excess of mobilization from natural deposits, atmospheric deposition, and small-scale uses of the metal as an amalgamating agent in gold mining. PMID- 9419262 TI - Organochlorine and heavy metal residues in breast muscle of known-age thick milled murres (Uria lomvia) from the Canadian Arctic. AB - Thick-billed murres (Uria lomvia) originating from breeding colonies in the Canadian Arctic were collected on their wintering grounds off the coast of Newfoundland. Murres had been previously banded such that the age of each bird could be determined upon collection. This allowed us to explore the possible relationships between age and contaminant levels in the thick-billed murre. Samples of breast muscle were analyzed for organochlorines (chlorobenzenes, hexachlorocyclohexanes, DDTs, chlordanes, mirex, dieldrin, and PCBs) and metals (selenium, cadmium, mercury, and lead). Levels of both organochlorine and metal residues were sufficiently low so that toxic effects were unlikely. First-year birds contained lower levels of DDTs, mirex, dieldrin, and PCBs compared with older birds, reflecting lower levels of contamination of these compounds in food chains at breeding colonies located at higher latitudes. Higher levels of chemical residues in older birds may reflect greater direct input of those organochlorines into the wintering grounds via the highly contaminated St. Lawrence River. Levels of chlorobenzenes, hexachlorocyclohexanes, and chlordanes, which reflect atmospheric deposition, were not detected at higher levels in older birds. Of the metals, only cadmium was detected at higher levels in older birds. PMID- 9419263 TI - Age differences in metals in the blood of herring (Larus argentatus) and Franklin's (Larus pipixcan) gulls. AB - Concentrations of heavy metals and selenium were measured in the blood of adult and young herring (Larus argentatus) and Franklin's (Larus pipixcan) gulls collected during the same breeding season in colonies in the New York Bight and in northwestern Minnesota, respectively. Concentrations were expected to be higher in young herring gulls collected in an urban, industrialized area, compared to young Franklin's gulls collected in a relatively pristine prairie marsh. Exposure is similar for the fledgling and adult gulls because by the time the blood of young gulls is drawn both adults and young have been eating foods from the surrounding region for two months; leading to the prediction that metal levels should be similar in adults and young. However, young Franklin's gulls had significantly higher levels of arsenic, cadmium, and manganese than adults; adults had significantly higher levels of mercury and selenium. Young herring gulls had significantly higher concentrations of arsenic and selenium, but lower levels of lead than adult herring gulls. Interspecific comparisons indicated that young Franklin's gulls had significantly higher levels of cadmium than young herring gulls, and adult Franklin's gulls had higher levels of selenium and chromium than adult herring gulls, but for all other comparisons herring gulls had higher levels of metals in their blood. Young herring gulls chicks had higher arsenic, manganese, and selenium levels and lower cadmium and lead levels in 1993 than in 1994. Overall, the levels in the two species were usually within an order of magnitude. PMID- 9419264 TI - A comparison of mercury levels in feathers and eggs of osprey (Pandion haliaetus) in the North American Great Lakes. AB - Osprey (Pandion haliaetus) eggs and chick feathers were collected for mercury analysis from nests at four Great Lakes study areas in Ontario (three "naturally formed" lakes in southern Ontario and one reservoir in northern Ontario) and two New Jersey study areas in 1991-1994. Adult osprey feathers were sampled from three Great Lakes study areas in 1991. Feathers sampled from chicks (approximately 28-35 days old) appear to be better indicators of local contaminant conditions since spatial patterns of mercury in known prey, yellow perch (Perca flavescens), also collected in these areas, were more similar to chick feathers than to eggs. Mercury levels were less variable in chick feathers than in eggs. Estimates of biomagnification factors using prey of known size at these areas were also less variable in feathers than in eggs. At naturally formed lakes, no significant correlation in mercury levels between eggs and chick feathers from the same nest was apparent, suggesting that the source of mercury contamination was not the same in these two tissues: mercury levels in eggs reflect mercury acquired on the breeding grounds, wintering grounds, and migratory route; mercury levels in chick feathers reflect local dietary conditions on the breeding grounds. Mercury levels in both osprey eggs and chick feathers were higher at the Ogoki Reservoir than at naturally formed lakes. Adult osprey feathers had higher mercury concentrations than chick feathers. Mercury levels in osprey eggs, chick feathers, and adult feathers did not approach levels associated with toxic reproductive effects. PMID- 9419265 TI - Blood levels of DDT and breast cancer risk among women living in the north of Vietnam. AB - A positive association has been reported between elevated tissue organochlorines (p,p'-DDT/p,p'-DDE, PCBs, dioxins) and breast cancer in some case-control studies and occupational cohort studies. We previously reported high serum levels of p,p' DDT and its metabolite p,p'-DDE in women living throughout Vietnam. We report here the results of a small hospital-based case-control study examining the association between blood levels of p,p'-DDT/p,p'-DDE and the risk of invasive breast cancer among residents of the north of Vietnam-an area where insecticides such as p,p'-DDT have been heavily used in the recent past. The study was conducted among patients admitted to a single hospital in the capital city of Hanoi in 1994. Study subjects were 21 women newly diagnosed with invasive adenocarcinoma of the breast, who served as cases, and 21 women of similar age with fibrocystic breast disease, who served as controls. No increase was evident in the relative risk of breast cancer with increasing tertiles of serum concentration of the compounds of interest, even after adjustment for major potential confounders, such as age at menarche, parity, history of lactation, and body weight. These results suggest that recent and past exposure to p,p'-DDT does not play an important role in the etiology of breast cancer among women living in a country with a tropical climate where insecticide use for mosquito control is common. PMID- 9419266 TI - Toxicity of inorganic compounds in the Spirotox test: a miniaturized version of the Spirostomum ambiguum test. AB - The Spirostomum ambiguum toxicity test has been intensively studied in the Department of Environmental Health Sciences, Warsaw University of Medicine for the last 5 years. The purpose of the present work was to develop and evaluate a miniaturized microplate version of the test, called the Spirotox test, and to estimate the toxicity of selected inorganic compounds to the Spirostomum ambiguum. The test was carried out in conventional 24-well (6 x 4) polystyrene multiwell plate. Preliminary test was one control and 11 toxicant concentrations with two duplicates. Definitive test was one control and five toxicant concentrations with three duplicates per concentration. Dilution of the sample was made directly in the plate. Toxicity series of heavy metals based on 24-h LC50 may be established as follows: Cu > Ag > Hg > Cr > Cd > Zn > Ni > Pb > Co > Mn. The series may be divided into four classes: extremely toxic: below 0.1 ppm (Cu, Ag, Hg); very toxic: 0.1-1.0 ppm (Cr, Cd, Zn, Ni); toxic: 1.0-10 ppm (Pb, Co); and low toxic: above 10 ppm (Mn). Anions were much less toxic to S. ambiguum than cations. Using the same classification, only cyanide (CN) was toxic, other anions were low toxic. Toxicity series based on 24-h LC50 may be established as follows: CN > SeO3 > Cr2O7 > NO2 > S2O3 >WO4 > BO3. PMID- 9419267 TI - Liquid-gas partitioning of the gasoline oxygenate methyl tert-butyl ether (MTBE) under laboratory conditions and its effect on growth of selected algae. AB - The partitioning of the widely used gasoline additive methyl tert-butyl ether (MTBE) between liquid growth media and gaseous phase was measured daily under laboratory conditions to determine how closely dissolved MTBE concentrations matched nominal concentrations. Total (gaseous and dissolved) MTBE averaged across 6 days for 29.6, 503.2, and 1005.7 mg L-1 MTBE treatments were 89.9, 90.3, and 73.0% of nominal, respectively, and mean dissolved MTBE in these same treatments were 74.6, 73.8, and 69.6% of total MTBE, respectively. This suggests that dissolved MTBE concentrations can vary substantially from nominal. The effect of MTBE on the growth of selected algae was also evaluated under laboratory conditions. Three unicellular algae, Selenastrum capricornutum (Chlorophyta), Navicula pelliculosa (Bacillariophyta), and Synechococcus leopoliensis (= Anacystic nidulans, Cyanophyta = Cyanobacteria), representative of three taxonomic groups, were used as test organisms. Toxicity tests were acute and increase in cell number was used as an indicator of growth. Algal species were exposed by injection of MTBE into sealed vessels containing defined liquid growth media. The growth of N. pelliculosa and S. leopoliensis was negatively affected at nominal 2400 mg L-1 MTBE, whereas the growth of S. capricornutum was negatively affected at nominal 4800 mg L-1 MTBE and positively affected at nominal 600 mg L-1 MTBE. The differential sensitivity of the growth of these representative species suggests that MTBE may alter algal community composition in the natural environment. PMID- 9419268 TI - Influence of water hardness on accumulation and elimination of cadmium in two aquatic mosses under laboratory conditions. AB - This study investigated the effect of water hardness on the accumulation and elimination of cadmium (Cd) by two aquatic mosses, Fontinalis dalecarlica and Platyhypnidium riparioides, under laboratory conditions. The two mosses were exposed to nominal Cd concentrations of 0, 0.8, 2, and 10 microg . L-1, which includes the concentration range generally found in nature. The influence of three levels of water hardness (very soft: 11.7 mg . L-1; soft: 44.2 mg . L-1; and hard water: 92.3 mg . L-1 as CaCO3) was measured while maintaining the alkalinity and pH constant during the 28-day exposure. The Cd accumulation by the aquatic mosses was rapid, showing the potential of accumulation and the sensitivity of this biomonitor. Even if the actual Cd concentration in the water was low (concentration <0.15 microg . L-1 to 6.82 microg . L-1 of Cd), the uptake of Cd was very fast and mostly linear. This study was conducted in water hardness comparable to that found in the Canadian shield (hardness was <100 mg . L-1 as CaCO3). When the actual Cd concentration in the water was as high as 6.82 microg . L-1, the uptake of Cd was mostly linear and the steady state condition was not reach. Accumulation rates of Cd were significantly different when the mosses were in very soft (11.7 mg . L-1) as compared to hard water (92.3 mg . L-1 as CaCO3). The elimination of Cd followed a very slow process for the two species studied. The elimination rates of Cd from the mosses were not influenced by water hardness. PMID- 9419269 TI - Alteration of [14C]-testosterone metabolism after chronic exposure of Daphnia magna to tributyltin. AB - Tributyltin (TBT) is a marine biocide that has been shown to alter the activity of cytochrome P450 monooxygenases and elicit toxicity indicative of androgenization in some species. The present study was conducted to determine whether TBT altered P450-, reductase-, and transferase-mediated testosterone metabolic processes in Daphnia magna at sublethal exposure concentrations. Two generations of daphnids were continuously exposed for 21 days to nominal TBT concentrations ranging from 0.31 to 2.5 microg/L TBT. The highest TBT concentration (2.5 microg/L) was lethal to 60% of the exposed organisms. Lower TBT concentrations elicited no adverse effects on molting or reproduction of the daphnids. No differences were observed in the response of the first- and second generation daphnids to the toxicity of TBT. The ability of daphnids to metabolize [14C]-testosterone in vivo was assessed following exposure of each generation to TBT. Production of hydroxylated, reduced/dehydrogenated, and glucose-conjugated metabolites of testosterone were all elevated following exposure of both generations to 1.25 microg/L TBT. These findings indicate that, under these conditions, TBT elicits no discernible effects on molting and reproduction of daphnids at sublethal concentrations, and testosterone metabolism is enhanced at concentrations approaching those that are lethal to organisms. Alterations of steroid metabolism by xenobiotics can be used as a more sensitive indicator of sublethal exposure in daphnids than reproductive endpoints. PMID- 9419270 TI - Application of a benthic euryhaline amphipod, Corophium sp., as a sediment toxicity testing organism for both freshwater and estuarine systems. AB - The use of an as-yet-undescribed euryhaline Corophium sp. amphipod as a sediment toxicity testing organism was assessed. The species was found to be ubiquitous in many tidal areas of the Hawkesbury River catchment. The salinity of habitat sites ranged from 0.1 to 24 ppt, sediment total organic carbon (TOC) ranged from 0.4% to 3.5%, and the fines content (< 63 micron particle size) of the sediment ranged from 4.3% to 47.6%. Monitored populations ranged from a density of 59 to 6622 individuals per m2, with freshwater sites with a sediment fines content greater than 20% having the highest population densities. The sensitivity of the Corophium sp. was assessed by using copper chloride and ammonium chloride as reference toxicants in a 96-h static water-only test and a 10-day static sediment test. The LC50 for copper in freshwater-only exposures was 80 to 86 microg/L, using adult animals collected from the field. In contrast, the LC50 for copper in freshwater sediment and the sediment pore water were 840 mg/kg (dry weight) and 99 microg/L, respectively. The LC50 for ammonia (total) in freshwater-only at pH 7 was 5.5 mg/L. In contrast, the LC50 for ammonia (total) in freshwater sediment and the sediment pore water were 110 mg/kg (dry weight) and 6 mg/L, respectively. Laboratory cultures of 5 per thousand to 15 per thousand salinity were optimal for supporting the release of juveniles. Juveniles collected from laboratory cultures had a LC50 for copper in 5 per thousand and 10 per thousand salinity of 9 microg/L and 28.5 microg/L, respectively, in water-only exposures. The juveniles would be suitable for use in the development of a chronic sediment toxicity test with growth as the endpoint. PMID- 9419271 TI - Biotic factors modifying acute toxicity of aqueous cadmium to estuarine amphipod Leptocheirus plumulosus. AB - A 96-h exposure to aqueous cadmium (Cd) is the recommended reference toxicity test for 10-day sediment bioassays with the estuarine amphipod, Leptocheirus plumulosus (US EPA 1994). This water-only test was used to assess the influence of organism size, sex, and nutritional status on the sensitivity of laboratory cultured L. plumulosus to Cd. In addition, the response of field-collected amphipods was compared to similarly sized laboratory animals to assess potential seasonal changes in Cd sensitivity. Lipid content of test organisms was measured in these seasonal experiments and those evaluating effects of nutritional status because of its potential as an indicator of physiological condition. LC50 values of laboratory animals size-sorted on nested 500-, 710-, and 1000-micron mesh sieves, increased with size class: 0.36, 0.65, and 0.88 mg Cd/L, respectively. Gravid females were less sensitive than males or mature females to aqueous Cd. Studies on the influence of the molt cycle on Cd toxicity indicated enhanced sensitivity of immediate postmolt animals that may explain some of the observed differences in Cd tolerance. Nutritional effects were investigated by comparing the sensitivity of fed and starved laboratory-reared amphipods. Starved juveniles and adults were significantly smaller than their fed counterparts and exhibited a 28-43% reduction in lipid content, respectively. However, comparison of LC50 values indicated no significant differences in sensitivity to Cd between starved and fed juveniles (0.23 vs 0.30 mg Cd/L) or adults (0.37 vs 0.52 mg Cd/L). Field collected amphipods were typically more sensitive to Cd than laboratory animals, regardless of the season, although their lipid content varied, ranging from 6.6% in August to 13.7% in November. Results are discussed with respect to the use and interpretation of toxicity tests with this species. PMID- 9419273 TI - Trace metals and antioxidant enzymes in gills and digestive gland of the Mediterranean mussel Mytilus galloprovincialis. AB - A seasonal variability of trace metal concentrations and antioxidant enzymes was observed in gills and digestive gland of the Mediterranean mussel Mytilus galloprovincialis from both a polluted and a nonpolluted population. Trace metals (As, Cu, Fe, Mn, Pb, and Zn) exhibited, in both organs, maximum values in later winter-early spring followed by a progressive decrease during the summer. While in the gills this behavior probably reflects a different bioavailability of metals, in the digestive gland it is influenced mainly by the progressive infiltration of the organ by gonadic tissues during gametogenesis. Metals, as other pollutants, are known to influence the oxidative status of these organisms and antioxidant enzymes have been often proposed as biomarkers of exposure to contaminants. In this respect, it was of interest to compare the variations of these biochemical parameters with those of metal levels in two mussel populations from a polluted and a nonpolluted site, respectively. The biochemical parameters examined included the level of glutathione and the activity of the following glutathione dependent and antioxidant enzymes: glyoxalase I, EC 4.4.1.5; glyoxalase II, EC3.1.2.6; glutathione S-transferases, EC 2.5.1.18; glutathione reductase, EC 1.6.4.2; Se-dependent, EC 1.11.1.9 and Se-independent, EC 2.5.1.18 glutathione peroxidases; catalase, EC 1. 11.1.6; superoxide dismutase, EC 1.15.1.1. Seasonal variations of trace metals did not appear to influence those of biochemical parameters, which generally showed an opposite trend with higher enzymatic activities in summer when trace metal concentrations were lower. The effects of metals on antioxidant enzymes were more evident when the two mussel populations were compared. In particular, organisms from the polluted site showed lower levels of glutathione and higher enzymatic activities of glyoxalase I even though the magnitude of these differences was not constant during the year. Moreover, native mussels from both the polluted and control populations exhibited limited differences in the activities of glutathione reductase, glutathione peroxidases, catalase, and superoxide dismutase, suggesting the possibility of some biochemical adaptation in organisms from chronically polluted environments. PMID- 9419272 TI - The influence of fresh water pollutants and interaction with Asellus aquaticus (L.) on the feeding activity of Gammarus pulex (L.). AB - The feeding response of juvenile amphipod Gammarus pulex (L.) was investigated following exposure to freshwater pollutants. The method employed is nondestructive, provides a rapid indication of the status of groups of individuals, and is based on a time-response analysis of the consumption of the eggs of Artemia salina and the determination of median feeding times or FT50s. The feeding activity of juvenile G. pulex was found to be a sensitive response criterion for use in assessing the sublethal toxicity of copper, lindane, and 3,4 dichloroaniline (3,4-DCA). Reductions in gammarid feeding activity were identified following 96 hours exposure at 12.1 microg/L copper or 8.4 microg/L lindane and 240 hours exposure at 918 microg/L 3,4-DCA. However, a significant increase was observed in the feeding rate of gammarids that had been exposed for 240 h at 0.09 microg/L lindane in comparison with control values. The increase in feeding rate may be interpreted as a possible stimulatory effect associated with the toxicant action of lindane. Increases in gammarid feeding activity were not determined during the experiments conducted with either copper or 3,4-DCA. A sustained reduction in G. pulex feeding rates may cause growth inhibition and impaired reproduction which have previously been identified as sublethal responses of other freshwater organisms exposed to comparable concentrations of lindane, 3,4-DCA, or copper. The feeding bioassay was also used as a tool in an investigation of species interactions in toxicant systems. The feeding responses of G. pulex, which had been maintained in the presence of Asellus aquaticus (as interacting pairs) and exposed to a range of concentrations of lindane or 3,4 DCA, were recorded and compared. The findings illustrate the complex nature of test systems that integrate the stresses of toxicant and competition. In the lindane test system a reduction in gammarid feeding activity was observed following a 96-h exposure with A. aquaticus at 3.8 and 6.0 microg/L lindane (mean measured concentrations). After a 240-h exposure period a decrease in feeding rate was recorded only for gammarids that had been exposed to 6.5 microg/L lindane, however exposure to very low concentrations of lindane (0.1 and 0.9 microg/L) resulted in a significant increase in gammarid feeding activity. In the experiment conducted with 3,4-DCA the calculation of median feeding times or FT50s of gammarids that had been exposed for 96 and 240 h in the toxicant treatment groups with A. aquaticus was largely precluded (in most groups less than 50% of the A. salina eggs were eaten). However, control group FT50 values were determined on each occasion the bioassay was performed, indicating that a substantial reduction in gammarid feeding activity had occurred in the majority of the 3,4-DCA treatment groups. PMID- 9419274 TI - Cadmium accumulation in liver and kidneys and hepatic metallothionein and glutathione levels in Rana ridibunda, after exposure to CdCl2. AB - Adult female Rana ridibunda were exposed to 200 ppm (mg/l) of cadmium (Cd as CdCl2) dissolved in water for 4, 10, and 30 days. The 96-h LC50 value for Cd was determined to 534 ppm. The concentration of Cd in the liver and kidneys and the effect of Cd on the concentrations of hepatic metallothionein (Mts) and glutathione (GSH), were estimated. Cd accumulated in the liver and kidneys in a time-dependent pattern, with the kidneys accumulating the heavy metal at higher rates after the tenth day of exposure. There was a strong positive correlation of Cd concentration between these two organs. The concentration of Mts and GSH increased with the increase of Cd concentration in the liver, following a time- and Cd-dependence pattern. The concentration of Mts and GSH was positively correlated with the concentration of Cd in the liver. Mts concentration was positively correlated with GSH concentration. PMID- 9419275 TI - Biomarker responses in whitefish (Coregonus lavaretus L. s.l.) experimentally exposed in a large lake receiving effluents from pulp and paper industry. AB - Physiological and biochemical biomarker responses were studied in juvenile whitefish (Coregonus lavaretus L. s.l.) exposed experimentally to effluent from the forest industry. The large study area (609 km2), Southern Lake Saimaa, in Southeast Finland, receives 330,000 m3 d-1 of biologically and 55,000 m3 d-1 of chemically treated effluents, discharged from two integrated elementary chlorine free (ECF) bleached kraft pulp and paper mills, from one ECF pulp mill, and from one mill producing unbleached pulp and cardboard. The assessment of exposure to effluent discharged from the mills was based on lake water chlorophenolics (CPs) and resin acids (RAs) measured in samples collected from the 22 experimental sites along the area. Despite the low levels of effluent constituents in the lake, they were still accumulated in detectable levels in fish bile, indicating an exposure to the bioactive compounds of effluents. In comparison to the reference area, a two- to four-fold increase in ethoxyresorufin O-deethylase (EROD) activity was observed in whitefish exposed in the vicinity (1-6 km) of all the mills. However, cytochrome P450 1A1 (CYP1A1) gene expression was increased in only one of the receiving areas, indicating higher sensitivity of the EROD activity in the present study. There were no statistically significant correlations between EROD activity and the ambient water concentrations of the CPs, the RAs, or effluent dilution expressed by water sodium concentration. Neither bile chlorophenolics nor bile resin acids showed a significant correlation with EROD. No significant changes in circulating reproductive steroids, 17beta-estradiol and testosterone, in juvenile whitefish were observed. The vitellogenin gene was expressed in the vicinity of the pulp mill discharging the most wood-derived compounds, i.e. resin acids and wood-sterols, including beta-sitosterol. No differences were observed in plasma immunoglobulin M, glucose, or lactate concentrations between the effluent sources. PMID- 9419276 TI - Concentrations and hazard assessment of organochlorine contaminants and mercury in smallmouth bass from a remote lake in the Upper Peninsula of Michigan. AB - Concentrations of PCBs, DDTs, toxaphene, chlordanes, dieldrin, and mercury were determined in smallmouth bass (Micropterus dolomieui) collected from Fumee Lake, a remote lake in the Upper Peninsula of Michigan. An ecological hazard assessment was conducted to determine potential impacts of contaminants on bald eagles and mink eating fish from this lake. Concentrations of organochlorines, except toxaphene, and mercury in smallmouth bass were similar to those found in fish from Lake Superior, where atmospheric inputs are the primary sources. Bioaccumulation was indicated by a positive correlation between fish weight and contaminant concentrations for organochlorines, while mercury concentrations did not appear to correspond predictably to body weight. Concentrations of mercury and PCBs in smallmouth bass were sufficiently great to be of concern regarding their consumption by eagles or mink. PMID- 9419277 TI - Heavy metal, organochlorine pesticide, and PCB residues in eggs and feathers of herons breeding in northern Italy. AB - We report on organochlorine pesticide and PCB concentrations in eggs of the little egret, Egretta garzetta, and the black-crowned night-heron, Nycticorax nycticorax, collected in 1993-1994, and on mercury, cadmium, and lead concentrations in feathers of 20-day-old nestlings collected from the same nests in 1994, from heronries near Pavia, northern Italy. Organochlorine pesticide and PCB residues were lower than those commonly associated with mortality and reduced reproductive success. As population levels of the species studied are not declining, these contaminants appear to have no significant adverse effect on reproduction in the heronries studied. DDE levels have decreased markedly in heron eggs since 1978. However, the presence of both DDT and beta-HCH, albeit at low levels, is notable, given that these compounds were banned in Italy in 1978 and 1988, respectively. Relatively high levels of Hg, Cd, and Pb in feathers suggest birds in their colonies are exposed to these contaminants, although both Cd and Pb may relate more to external than to internal contamination. PMID- 9419278 TI - Evaluation of early embryonic loss induced by tributyltin chloride in rats: phase and dose-dependent antifertility effects. AB - In our previous study, tributyltin chloride (TBTCl) on days 0-7 of pregnancy was found to produce implantation failure in rats. The objective of the present study was to determine the susceptible period for the antifertility effect of TBTCl in rats. Inseminated females were orally administered TBTCl at 8.1, 16.3, or 32.5 mg/kg on days 0-3 of pregnancy, or at 8.1, 16.3, 32.5, or 65.1 mg/kg on days 4-7 of pregnancy. Pregnancy outcome was determined on day 20 of pregnancy. Dosing with TBTCl on days 0-3 of pregnancy at 16.3 mg/kg and higher produced a significant increase in the rate of implantation failure. Dosing with TBTCl on days 4-7 of pregnancy caused a significant increase in the incidence of postimplantation loss at 16.3 mg/kg and higher in females with implantations. No increase in the incidence of fetal malformations was found in any TBTCl-treated groups. It could be concluded that the susceptibility to and manifestation of the antifertility effects of TBTCl vary with the gestational stage at the time of administration. PMID- 9419279 TI - Concentration of mercury in hair of indigenous mothers and infants from the Amazon basin. AB - Hair mercury concentration, as an indicator of mercury body load, was studied in 251 samples of indigenous women and children living in selected areas of the Amazonian region. The mothers or women of child-bearing age, either non-Indians or Indians, and their children were sampled along the Madeira River and in the Kayapo reservation (Fresco River), respectively. Among the sampled individuals there were mothers with infants less than 2 years old. Total mercury in hair was determined by cold vapor atomic absorption spectrometry after alkaline digestion. The distribution of hair mercury concentration greater than 10 microg/g occurred in 67.4% of non-Indian women and 25% of Indian women; overall only 1% of non Indian women had concentrations of hair mercury above 50 microg/g. In women of child-bearing age, the median and range of hair mercury concentration was 14.08 microg/g, and 0.8-94.7 microg/g for non-Indians, and 8.30 microg/g, and 0.8-13.3 microg/g for Indians. The correlation between maternal hair mercury and mercury in hair of infants (less than 2 years of age) still breast-feeding, was statistically significant only for non-Indians (r = 0.555 p < 0. 001). The correlation between length of breast-feeding and mercury concentration in infant's hair was significant for Indian children (r = 0.512; p = 0.029) but not for non-Indian children (r = 0.025; p = 0.832). A subsampling of 30 mothers had segmented hair analysis that showed a mean decrease of 20% in body burden during pregnancy, thus indicating the extent of placental transference of mercury to fetuses. PMID- 9419280 TI - FORUM: Bilateral Monopoly: A Market for Intercountry River Water Allocation AB - / Collaboration of countries with an aim to share fresh surface water resources promises to generate potential joint benefits. Unfortunately, existing agreements lack the perspective and capacity to produce any real action in efficient cross border water allocation. When that problem is encountered by any two adjacent countries claiming riparian rights to the same watercourse, this paper suggests that apossible solution to be examined is a water market. This market requires the relevant countries to engage in a bargaining process as described in the theory of bilateral monopoly. The bargaining process should determine both the water quantity to be transferred and the price to be paid. However, there has to be a fair allocation of the joint benefits resulting from the transfer for a sustainable price solution. As an empirical illustration, the paper examines the case of river Nestos shared by Bulgaria and Greece in the southern Balkans. A net revenue function quadratic in water is specified and estimated using scarce data on three agricultural crops in Greece. Sensitivity analysis on the size and distribution of the net benefits is also performed.KEY WORDS: Bilateral agreements; Water markets; Efficient allocation PMID- 9419281 TI - PROFILE: Impending Recovery of Kirtland's Warbler: Case Study in the Effectiveness of the Endangered Species Act AB - / The Endangered Species Act (ESA) has received a large amount of criticism in recent years by conservative landowners and others who believe that it has infringed on property rights. It also has been criticized by those who think it has been costly and ineffective in reaching its goal of preventing extinction and recovering species. Recent evidence, however, shows that the ESA has stabilized or increased the populations of over a third of the listed species. In addition, its chief administrator, the US Fish and Wildlife Service, has been increasingly flexible in implementing the ESA. After reviewing the administrative machinery of the ESA, this paper provides a case study of one endangered species, the Kirtland's warbler (Dendroica kirtlandii). This particular recovery program actually began before passage of the federal ESA, when biologists alerted the Michigan Department of Natural Resources of the perilously low population of this bird, which only breeds under jack pine (Pinus banksiana) trees in Michigan. By the time an ESA Recovery Team was formed for this bird in 1975 (the first such team created under the ESA), a legacy of consensus and interagency cooperation was well established. This has led to successful efforts at habitat management and control of its nest parasite, the brown-headed cowbird (Molothrus ater). While the Kirtland's warbler is not yet recovered, its population is near an all time high, and its recovery is possible within the next decade. When (and if) this happens, it will be clearly attributable to this successful model of federalism for natural resources management.KEY WORDS: Endangered species; Kirtland's warbler; Brown-headed cowbird; Jack pine; Endangered Species Act PMID- 9419282 TI - Sources of Deforestation in Tropical Developing Countries AB - / Key causes of tropical deforestation are investigated using cross-sectional data for 90 developing countries for the period 1981-1990. Regression results reveal that deforestation is associated with both development and scarcity. Deforestation accelerates with expanding infrastructure, trade, debt, investment in the human capital base, and resource-based economic expansion. On the other hand, absolute and relative scarcities-manifested by growing population pressures, food and land shortages, fuelwood dependency, and inequalities in access to land-are also key factors explaining forest loss. Thus, results point to a fundamental environmental conundrum: Development is required if countries are to alleviate scarcity-driven forms of forest exploitation but is itself a major cause of deforestation. Can countries balance development goals with forest protection? Setting aside the issue of its practical realization, the paper concludes that forest sustainable development cannot be achieved by implementing simple technical improvements in land-use practices alone. Securing the foundations for the sustainability of the forest base will require that countries address the underlying social processes driving tropical forest loss as well.KEY WORDS: Tropical deforestation; Developing countries; Rural land-use practices; Development; Scarcity. PMID- 9419283 TI - A Knowledge-Based Systems Approach to Design of Spatial Decision Support Systems for Environmental Management AB - / This paper describes a framework for designing spatial decision support systems for environmental management using a knowledge-based systems approach. An architecture for knowledge-based spatial decision supportsystems (KBSDSS) is presented that integrates knowledge-based systems with geographical information systems (GIS) and other problem-solving techniques. A method based on spatial influence diagrams is developed for representation of environmental problems. The spatial influence diagram provides an interface through which knowledge-based systems techniques can be applied to build capabilities for problem formulation, automated design, and execution of a solution process. In addition to the flexibility and developmental advantages of knowledge-based systems, the KBSDSS incorporates expert knowledge to provide assistance for structuring spatial influence diagrams and executing a solution process that automatically integrates the GIS, data base, knowledge base, and different types of models. The framework is illustrated with a system, known as the Islay Land Use Decision Support System (ILUDSS), designed to assist planners in strategic planning of land use for the development of the island of Islay, off the west coast of Scotland.KEY WORDS: Geographical information systems; Spatial decision support systems; Knowledge based systems; Spatial influence diagrams; Environmental management PMID- 9419284 TI - Valuation of Ecological Resources and Functions AB - / Ecological resources are natural resources that provide certain necessary but overlooked system maintenance functions within ecosystems. Environmental economics is in search of an appropriate analysis framework to determine economic values of such resources. This paper presents a framework that estimates and compiles the components of value for a natural ecosystem. The framework begins with the ecological processes involved, which provide functions within the ecosystem and services valued by humans. We discuss the additive or competive nature of these values, and estimate these values through conventional and unconventional techniques. We apply the framework to ecological resources in a shrub-steppe dryland habitat being displaced by development. We first determine which functions and services are mutually exclusive (e.g., farming vs soil stabilization) and which are complementary or products of joint production (e.g., soil stabilization and maintenance of species). We then apply benefit transfer principles with contingent valuation methodology (CVM), travel cost methodology (TCM), and hedonic damage pricing (HDP). Finally, we derive upper-limit values for more difficult-to-value functions through the use of human analogs, which we argue are the most appropriate method of valuation under some circumstances. The highest values of natural shrub-steppe habitat appear to be derived from soil stabilization.KEY WORDS: Natural resource economics; Ecological economics; Ecological resources; Shrub-steppe; Environmental valuation; Cost; Benefit; Value PMID- 9419285 TI - RESEARCH: An Ecoregional Approach to the Economic Valuation of Land- and Water Based Recreation in the United States AB - / This paper describes a framework for estimating the economic value of outdoor recreation across different ecoregions. Ten ecoregions in the continental United States were defined based on similarly functioning ecosystem characters. The individual travel cost method was employed to estimate recreation demand functions for activities such as motor boating and waterskiing, developed and primitive camping, coldwater fishing, sightseeing and pleasure driving, and big game hunting for each ecoregion. While our ecoregional approach differs conceptually from previous work, our results appear consistent with the previous travel cost method valuation studies.KEY WORDS: Recreation; Ecoregion; Travel cost method; Truncated Poisson model PMID- 9419286 TI - Linking the Wilderness Perception Mapping Concept to the Recreation Opportunity Spectrum AB - / This paper examines the application of a wilderness perception mapping (WPM) approach to enhancingthe recreation opportunity spectrum (ROS). WPM provides a greater understanding of the multiple relationships between wilderness environments and wilderness user experience. The results from a case study indicate that different recreation opportunity settings provide experiential conditions of wilderness for different groups. Wilderness perception mapping complements the use of ROS and can be applied to wilderness management.KEY WORDS: Geographical information systems; Recreation opportunity spectrum; Outdoor recreation planning; Wilderness management; Wilderness perception mapping; New Zealand PMID- 9419287 TI - Changes in Use of Three Oregon, USA, Wildernesses AB - / Several studies have suggested that use of wildernesses within national forests is declining. Data from three wildernesses in Oregon (based on mandatory permits and voluntary registration) indicate that between 1976 and 1991 in two areas and between 1980 and 1993 in another area the number of recreational visitor days (RVDs) has declined between 23% and 63%. However, the number of visits has remained roughly stable in one area and more than doubled in the other two. The disparity between the two measures of use may be the result of shifts from mainly overnight to mainly day use in these wildernesses. Day use has increased from about 30% to over 60% of all visits. Trends are different enough in each wilderness to warrant independent analyses over time. Management implications of using different measures of use as well as observed trends are discussed.KEY WORDS: Wilderness management; Wilderness use; Trends PMID- 9419288 TI - Environmental Management of Human Waste Disposal for Recreational Boating Activities AB - / A methodology to estimate the number of pump-out facilities and dump stations required to service human waste disposal for recreational power boating activities in Pennsylvania during the 1994 boating season is described. Study results suggest that a total of 39 additional pump-out stations and 13 dump stations may be required on seven major waterbodies: The Three Rivers Area, Lake Erie/Presque Isle Bay, Raystown Lake, the Susquehanna River, the Delaware River, Lake Wallenpaupack, and the Kinzua Reservoir. Suggestions for improving the methodology are provided. KEY WORDS: Human waste; Recreation; Power boating; Waste facilities; Waste disposal; Pennsylvania PMID- 9419289 TI - A Path Model of Whitewater Boating Satisfaction on the Cheat River of West Virginia AB - / Recreation satisfaction is a complex psychological construct that is difficult to define and measure. Recent approaches suggest that overall satisfaction may be a function of multiple satisfactions derived from specific elements of a recreation experience such as the situational characteristics of a recreation setting or activity and the recreationist's subjective evaluations of the experience. In this paper, a path model of whitewater boating satisfaction was tested using data from a survey of 1210 commercial and 111 private boaters on the Cheat River of West Virginia. The pathmodel included the direct and mediating effects of situational variables and the subjective evaluations of boaters and explained 52% and 54% of the variation in satisfaction of commercial and private boaters, respectively. Factors related to the satisfaction of both groups included a composite variable representing opportunities for challenge, excitement, and skill testing on the river trip; water flow levels; and crowding perceptions. In combination, water flow level and boater's perceptions of opportunities to experience challenge, excitement, and test boating skills were the most important variables for explaining satisfaction of both groups. Additional factors affecting commercial, but not private, boater satisfaction included the motive of escaping the usual demands of life and a social interaction variable. Among private boaters, perceptions of the environmental conditions also contributed to overall satisfaction. The results support the multiple satisfaction approach of previous research. River management implications are discussed.KEY WORDS: Whitewater; River recreation; Satisfaction PMID- 9419290 TI - Long-Term Growth Enhancement of Baldcypress (Taxodium distichum) from Municipal Wastewater Application AB - / Tree ring analysis was used to document the long-term effects of municipal wastewater on the growth rate of baldcypress [Taxodium distichum (L.) Rich.]. The study site, a swamp in St. Martin Parish, Louisiana, has received municipal wastewater for the last 40 years. Growth chronologies from 1920 to 1992 were developed from cross-dated tree core samples taken from treated and control sites with similar size and age classes. Mean diameter increment (DINC) and mean basal area increment (BAI) chronologies were constructed separately for each stand. These chronologies were then summarized by tree and stand into seven nine-year intervals resulting in three pretreatment intervals from 1926 to 1952 and four treatment intervals from 1953 to 1988. Significant differences in growth response between sites showed a consistent pattern of growth enhancement in the treated site coincident with the onset of effluent discharge. The ratio of treated to control baldcypress growth rates (computed from DINC) averaged 0.74 during the pretreatment period and 1.53 during the treatment period. Over the period of study, control DINC decreased from 77 mm to 29 mm/nine-year interval, while treatment DINC increased slightly from 40 mm to 47 mm/nine-year interval. Control BAI did not increase significantly and averaged 192 cm2/nine-year interval. There was a significant increase in treatment BAI from 129 to 333 cm2/nine-year interval over the period of record. These results clearly demonstrate sustained long-term baldcypress growth enhancement throughout 40 years of municipal effluent discharge.KEY WORDS: Taxodium; Tree-ring analysis; Dendroecology; Wetlands; Natural wetland wastewater treatment; Long-term growth enhancement PMID- 9419291 TI - ENVIRONMENTAL AUDITING: The Fish Fauna of the Doubs River Prior to Completion of the Rhine-Rhone Connection AB - / The part of the Doubs River between Montbeliard and Dole (France), i.e., downstream from the confluence with the Allan River, will be affected by the Rhine- Rhone connection project. In order to improve the understanding of the Doubs ichthyofauna, aquatic environments of the Doubs were sampled by electrofishing. Fish diversity and the presence of some rheophilic species demonstrated the good ecological quality of some stretches of the Doubs. This quality was due to alternating areas with very diversified aquatic environments (riffles, islands and side-arms, backwaters) and a considerable range of flow velocities. The differences in the structure of the fish communities of the different types of aquatic environments were more qualitative (fish species) than quantitative (number of species and number of fish). However, the mean number of fish was statistically lower in the canals (Freycinet canal and channelized part of the Allan River) than in the main course and in the backwaters. The natural parts of the Doubs (unnavigable reaches) showed the most diversified environmental structure and had the most rheophilic fish communities. Thus, the rheophilic species were well represented, but they proved also the most vulnerable to river regulation. However, the most abundant fishes throughout the Doubs River were generalists with no special requirements for food sources or spawning substrate.KEY WORDS: Fish communities; Regulation; Restoration; Floodplain; Large ship canal; Doubs River PMID- 9419292 TI - Isotopes, Wool, and Rangeland Monitoring: Let the Sheep Do the Sampling AB - / Stable carbon isotope analyses of wool staples provided insight into the vegetation consumed by sheep at a temporal resolution not previously studied. Contemporary Australian and historic South African samples dating back to 1916 were analyzed for their stable carbon isotope ratio, a proxy for the proportion of C3 and C4 plant species consumed by animals. Sheep sample vegetation continuously throughout a year, and as their wool grows it integrates and stores information about their diet. In subtropical and tropical rangelands the majority of grass species are C4. Since sheep prefer to graze, and their wool is an isotopic record of their diet, we now have the potential to develop a high resolution index to the availability of grass from a sheep's perspective. Isotopic analyses of wool suggest a new direction for monitoring grazing and for the reconstruction of past vegetation changes, which will make a significant contribution to traditional rangeland ecology and management. It is recommended that isotopic and other analyses of wool be further developed for use in rangeland monitoring programs to provide valuable feedback for land managers.KEY WORDS: Carbon isotope ratios; Vegetation change; Rangelands; Monitoring; Wool PMID- 9419293 TI - Synthetic Analysis for Extracting Information on Soil Salinity Using Remote Sensing and GIS: A Case Study of Yanggao Basin in China AB - / This paper reports the experience of extracting information on the salinity of soil and offers a method of synthetic analysis. The experimental areas for analysis are located in Yanggao Basin, Shanxi Province, China. The types of soil are mainly meadow soil and salinized meadow soil. The method of synthetic analysis of salinity uses a geographic information system (GIS) as a tool, building a basic saltwater analysis model of saline soil and adjusting the result with expert experience after computer processing. The method of feature extraction has been used for remotely sensed data. An optimum combination of features has been determined and, after comparing several combinations in the Yanggao region, an improved result has been obtained after Kauth-Thomas (K-T) transformation. For precise quantitative analysis of the salinization, not only Thematic Mapper (TM) remote sensing data, but also two forms of non-remote sensing data are needed: depth of groundwater and mineralization rate of groundwater according to the theory of genesis of soil. For the analysis of synthetic compounded multisources, a generalized Bayes classification is used after overlay, matching, and related coefficients have been determined. On the premise that various information sources are independent, global membership functions with probability are used to combine various pieces of information in order to apply them directly to the pixels and classifications of soil salinity. The experiment indicates that this analytical method is sound because of the increased speed of processing and its simplicity and improved precision of classification of salinity. Finally, it is necessary to examine and adjust the factors using expert intelligence. The experiment shows that synthetic analysis using the geographic information system can raise the precision of quantitative analysis of salinity, which has advantages for environmental monitoring and management.KEY WORDS: Salinity; Remote sensing; Thematic Mapper; Geographic information system; Classification PMID- 9419294 TI - Transperitoneal laparoscopic pelvic lymphadenectomy for gynecologic malignancies (I). Technique and results. AB - BACKGROUND: We reviewed the published experimental and clinical data, available in MEDLINE, and compared them to our experience in a university-affiliated tertiary medical center of obstetrics and gynecology in order to describe the accepted techniques and results of laparoscopic pelvic lymphadenectomy. METHODS: The procedure requires a four-port access laparoscopy. Dissection boundaries are similar to those for open surgery. RESULTS: Experimental and clinical comparative series have shown that the number of harvested lymph nodes is not significantly different for laparoscopy than for laparotomy. Several authors reported a learning curve, reflecting the surgeon's increasing accuracy with growing operative experience. Obesity and prior history of laparotomy are both factors that impact adversely on the number of nodes harvested and the complication rate. Otherwise, the number of residual nodes is similar for the two approaches. In both cases, it is low, resulting in a high sensitivity (95-100%). The complication rate is directly linked to the surgeon's experience and thus appears low for skilled laparoscopic operators. It is similar to that reported for open surgery. Anesthesiological complications have not been well assessed in the literature on laparoscopic lymphadenectomy. Operating time was longer than for laparotomy in all the series. Conversely, mean blood loss, duration of hospitalization, and recovery time were significantly decreased. Although intraoperative cost of the laparoscopic procedure is high in comparison with laparotomy, since the time of recovery appears shorter, total costs may be similar or even lower. CONCLUSION: We conclude that laparoscopic pelvic lymphadenectomy is a reliable and safe procedure for the evaluation and treatment of gynecologic cancers. PMID- 9419295 TI - Intermittent pneumatic sequential compression (ISC) of the lower extremities prevents venous stasis during laparoscopic cholecystectomy. A prospective randomized study. AB - BACKGROUND: Fifty patients were included in a prospective randomized trial to evaluate the efficacy of intermittent sequential compression (ISC) of the lower extremities in preventing venous stasis during laparoscopic cholecystectomy. METHODS: We treated 25 patients with (+ISC) and 25 without (-ISC) intermittent sequential compression. Peak flow velocity (PFV) and cross-sectional area (CSA) of the right femoral vein were measured by Doppler ultrasound before, during, and after capnopneumoperitoneum with 14 mm Hg. RESULTS: PFV was 26.4 (8.4) cm/s and CSA was 1.03 (0.23) cm2 before pneumoperitoneum was induced. During abdominal insufflation, PFV decreased to 61% of the baseline value in the (-ISC) group but remained unchanged in the (+ISC) group (t = 5.17, df = 42.8, p < 0.01). CSA was 1.06 (0.22) cm2 before insufflation. It increased to 118% of the baseline in the (-ISC) group and to 108% in the (+ISC) group (t = -1.55, df = 47.1, p = 0.13). PFV and CSA returned to baseline values within 5 min after abdominal desufflation. CONCLUSIONS: ISC effectively neutralizes venous stasis during laparoscopic surgery and may decrease the risk of post-operative thromboembolic complication. Therefore, it is recommended for all prolonged laparoscopic procedures. PMID- 9419296 TI - How often might a trans-cystic-duct stone extraction be feasible? AB - BACKGROUND: Although the sizes of the cystic duct and concomitant bile duct stones are fundamental in evaluating the possibility of a trans-cystic-duct approach as an alternative to cholangiotomy, no conclusive data are supplied in the reports on laparoscopic cholecystectomy. METHOD: The narrowest inner diameter of the cystic duct and the diameter of the largest concomitant bile duct stone are compared in a prospective study of 30 consecutive patients. RESULTS: The bile duct stones were smaller than the cystic duct in 14 patients, 47%, and of equal size in nine, 30%. They were larger than the cystic duct in the remaining seven patients, 23%, with a difference of only 1 mm in five patients and of 2 and 4 mm, respectively, in two. CONCLUSIONS: Physical conditions allowing a trans-cystic duct stone extraction were present in 23 of 30 patients and an attempt might have been possible after, for example, cystic duct dilatation in a further five. PMID- 9419297 TI - Evaluation of laparoscopic management of common bile duct stones in 220 patients. AB - BACKGROUND: The aim of this study was to evaluate the feasibility and results of laparoscopic management of common bile duct stones (CBDS). METHODS: From October 1990 to November 1996, 220 patients with CBDS have been managed laparoscopically. CBDS were suspected or diagnosed preoperatively in 130 patients (59.1%) and at intraoperative cholangiography (IOC) in 90 patients (40.9%). A transcystic duct extraction (TCDE) was attempted in 112 patients and a primary choledochotomy in 108 patients. RESULTS: TCDE was successful in 77 cases (68.8%). The 35 failures were treated by 29 laparoscopic choledochotomies, 1 intraoperative and 5 postoperative endoscopic sphincterotomies (ES). A choledochotomy was thus performed in 137 cases and was successful in 133 cases (97.1%). The four failures were managed by three laparotomies and one postoperative ES. The overall success rate was 95.5% (210/220). There was 4 deaths (0.9%) within the 1st postoperative month in ASA 3 patients and the morbidity rate was 9.1% (20/220). There were 7 residual stones (3.2%). CONCLUSIONS: Laparoscopic desobstruction of CBDS appears to be safe and effective and has the advantage to be a single-stage procedure. It could become in the future with refinement of instrumentation and skill of surgeons the best treatment for the majority of patients harboring CBDS. PMID- 9419298 TI - Follow-up of 161 unselected consecutive patients treated laparoscopically for common bile duct stones. AB - BACKGROUND: Aim was to study the incidence of recurrent ductal stones and of biliary strictures at follow-up after laparoscopic treatment of gallstones and common bile duct stones and to update the short-term results. METHODS: Ductal stones were proven in 161 patients of 1,975 (8.1%) undergoing laparoscopic cholecystectomy. Laparoscopic transcystic CBD exploration was the method of choice. If this was unsuccessful, laparoscopic choledochotomy was performed. After treatment, all patients were enrolled in a continued, ongoing follow-up study. RESULTS: Laparoscopic CBD exploration was completed in 157 cases (transcystic 107, choledochotomy 50). Retained stones occurred in eight patients (5%) and major complications (cystic duct leakage, hemoperitoneum) in six (3.8%); mortality occurred in one high-risk patient (0.6%). Follow-up available in 154 patients (two unrelated deaths) for a period of up to 62 months showed the occurrence of recurrent ductal stones in five cases (3.2%) and no signs of bile stasis, suggestive of ductal stricture, on the basis of clinical and laboratory findings. CONCLUSIONS: This prospective, ongoing follow-up study demonstrates that laparoscopic treatment of gallstones and common bile duct stones in unselected patients is feasible and safe. PMID- 9419299 TI - Prospective randomized comparison of laparoscopic ultrasonography using a flexible-tip ultrasound probe and intraoperative dynamic cholangiography during laparoscopic cholecystectomy. AB - BACKGROUND: We performed a prospective randomized comparison of laparoscopic intraoperative ultrasonography (LIOU) and dynamic intraoperative cholangiography (IOC) during laparoscopic cholecystectomy (LC). METHODS: LIOU and IOC were attempted in 518 consecutive patients scheduled for laparoscopic cholecystectomy. The order in which the diagnostic procedures were performed was randomly assigned. RESULTS: LIOU failed in two patients (0.4%), and there were 41 (7.9%) failed IOC. The common bile duct (CBD) was visualized reliably with both methods. Our patients showed sensitivities of 83.3% and 100% and specificities of 100% and 98.9%, with an overall accuracy of 99.2% and 98.9% for LIOU as compared to IOC for identifying unsuspected common bile duct stones. The time necessary for the examination was significantly shorter in LIOU than in IOC (7 versus 16 min). CONCLUSION: LIOU performed by experienced surgeons is a good and effective method to assess the CBD, including the neighboring structures of hepatoduodenal ligament. Using powerful, flexible-tip ultrasound probes, CBD exploration can be done in a longitudinal fashion, which is necessary for good anatomical clarity. A lack of adverse effects, shorter examination times, and lower costs are some of the advantages of this method. The most important advantage is the possibility of unlimited repetition, especially if there is difficulty identifying anatomic structures. In addition, there are some indications that LIOU has the potential to recognize major iatrogenic bile duct injuries. PMID- 9419300 TI - Early results of a prospective multicenter study on 500 consecutive cases of laparoscopic colorectal surgery. Laparoscopic Colorectal Surgery Study Group (LCSSG). AB - BACKGROUND: Prospective randomized multicenter studies comparing laparoscopic with open colorectal surgery are not yet available. Reliable data from prospective multicenter studies involving consecutive patients are also lacking. On the basis of the personal caseloads of specialized surgeons or of retrospective analyses, it is difficult to judge the true effectiveness of this new technique. This study aims to investigate the results of laparoscopic colorectal surgery in consecutive patients operated on by unselected surgeons. METHODS: This observational study was begun August 1, 1995, in the German speaking part of Europe (Germany and Austria) and 43 centers initially agreed to participate. All consecutive cases were documented. All data were rendered anonymous. Analysis was performed on an intention-to-treat basis. The study committee was blinded to the participating center. RESULTS: By the end of the 1st year, 500 patients (M:F ratio 0.83, mean age 62.9 years) had been treated by 18 centers; 269 operations were performed for benign indications and 231 for cancer (palliative and curative). Most operations were done on the distal colon or rectum. An anastomosis was performed in 84%, with an overall leakage rate of 5.3% (colon 3.6% and rectum 11.8%), which required surgical reintervention in 1.7%. The mean operating time was 176 min and showed a decreasing tendency over the period under study. The conversion rate was 7.0% and the overall complication rate 21.4%. The reoperation rate was 6.6%; the most common cause was bleeding. There was one ureteral lesion (0.2%), but urinary tract infections were fairly common (4.8%). A postoperative pneumonia was diagnosed in 1.6% of the cases. No thromboembolic complications were reported. The 30-day mortality rate was 1.4% and overall hospital mortality 1.8%. CONCLUSIONS: Laparoscopic colorectal operations are still rare (about 1% of all colorectal operations in Germany). Laparoscopic procedures are more common on the left colon and rectum than on the right colon. The surgical complication rate is acceptable, comparable with rates reported by others for open surgery. Cardiopulmonary and thromboembolic complications were rarely seen. Mortality and surgical morbidity rates do not differ significantly among participating centers. A learning curve, reflected by a shortening of the operating time and a somewhat lower conversion rate, was observed over the observation period. PMID- 9419301 TI - Indications for laparoscopic colorectal surgery. Results from the Medical Centre Alkmaar, The Netherlands. AB - BACKGROUND: Between November 1991 and May 1995, a series of laparoscopic colectomies were performed in our hospital. METHODS: Our main aim was to define more specifically the indications for laparoscopic colectomy. RESULTS: A total of 69 patients underwent laparoscopic surgery for benign polypoid colorectal disease (n = 10), inflammatory bowel disease (n = 24), and colorectal malignancy (n = 35). Of the latter group, four patients underwent a palliative procedure. The conversion rate of the whole group was 29%. The main reason to convert was infiltrative growth in inflammatory disease or cancer. Respectively, seven (10%) and 12 (17%) patients sustained complications in the perioperative and early postoperative phase. Two patients died perioperatively (3%). The mean hospital stay was 12 days. On follow-up, 11 patients had developed a stenotic anastomosis, which was successfully dilated in all cases. After 3 years, the survival rate according to Kaplan-Meier is 86%, 66%, 68%, and 0% for Dukes' A, B, C, and D color carcinoma, respectively. In one patient with a Dukes B carcinoma, port site metastases were found. CONCLUSIONS: Justifiable indications for laparoscopic colorectal surgery include (a) a benign polyp 20-50 cm from the anal ring; (b) mobile, inflammatory large bowel disease; (c) palliation in case of malignant disease, preferably of the left hemicolon. It remains to be proven that laparoscopic colectomy is superior and not just equivalent to open colectomy. This is especially true for resections of colorectal carcinoma with curative intent. Therefore a cost/benefit analysis should be performed in a prospective, randomized setting. PMID- 9419302 TI - Laparoscopic management of cystic disease of the liver. AB - BACKGROUND: Laparoscopic management of cystic disease of the liver, including severe polycystic disease, is evolving. METHODS: Wide unroofing, or "fenestration," as is required for a successful result in open cases, leads to complete resolution of the cysts. This can even occur in chronic cysts, with wide enough unroofing, given time. RESULTS: In polycystic disease, adequate fenestration of superficial, cysts allows deeper cysts to prolapse and be similarly fenestrated, thus reducing pressure effects on the liver and restoring normal function. CONCLUSION: However, because of the distortion of anatomy by this disease, it is important that an experienced liver surgeon perform such a complex procedure, as operative complications could be severe. PMID- 9419303 TI - Laparoscopic repair of a paraduodenal hernia. AB - Paraduodenal hernias have traditionally been treated by conventional laparotomy. We report the first case of a left paraduodenal hernia treated laparoscopically. A 44-year-old man was admitted with abdominal pain and nausea. Computed tomography and an upper gastrointestinal series with small-bowel followthrough showed accumulation of the small bowel on the left side of the abdomen. A laparoscopic repair was performed. The small bowel was observed beneath a thin hernia capsule. Approximately 1.5 m of jejunum was easily reduced into the abdominal cavity. The hernia orifice (5-cm diameter) was closed intracorporeally with five interrupted sutures. Good exposure of the operative field is critical to this procedure; poor exposure may limit the applicability of the laparoscopic approach. This minimally invasive operation is currently indicated in nonobstructive paraduodenal hernias, especially on the left. PMID- 9419304 TI - Laparoscopic resection of splenic artery aneurysm. AB - A new, lateral approach was used for the laparoscopic resection of splenic artery aneurysm. This approach was found to be convenient and straightforward. PMID- 9419305 TI - Videothoracoscopic resection of anterior mediastinal teratoma in a child. Report of a case. PMID- 9419306 TI - Two unusual cases of postcholecystectomy pain. AB - I report on two patients who were initially diagnosed with sphincter of Oddi dysfunction (S.O.D.) because of postcholecystectomy pain in the right upper quadrant; both had other causes of pain. One patient had an aberrant hepatic duct that drained into a remnant of the cystic duct resulting in formation of stones. The second patient had adhesions of the stomach to the liver with the ligamentum teres bowing across the antrum. Gastroenterologists and endoscopic surgeons should be aware of causes of postcholecystectomy pain that are unrelated to sphincter of Oddi dysfunction. PMID- 9419307 TI - Common bile duct T-tubes. A caveat and recommendations for management. AB - Operations on the common bile duct can result in severe long-term consequences. To prevent some of these complications, it is common practice to drain the biliary tree with a T-tube. The T-tube is usually removed 2 weeks after it was placed. There have been numerous reports of bile leak following T-tube removal in the literature. These leaks can result in bile ascites, biloma, or bile peritonitis. Control of bile leaks can be accomplished in a number of ways, including endoscopically or radiologically placed stents or drains and radiologic techniques to drain the fluid collections. We describe a novel technique that can be utilized at the time of T-tube removal that will allow immediate control of the bile leak and prevent the complications of bile accumulation within the peritoneal cavity. We have performed fluoroscopic removal of T-tubes on two patients and found no complications with the technique. We have successfully visualized the T-tube tract in both patients. The T-tube tract can be visualized at the time of T-tube removal in an effort to prevent the complications of tract disruption and subsequent bile leak. PMID- 9419309 TI - Laparoscopic surgery for splenic disorders. Lessons learned from a series of 64 cases. AB - Laparoscopic splenectomy (LS) has recently been gaining acceptance as an alternative to open splenectomy. However, several aspects, such as learning curve, residual splenic function, and management of large spleens, remain controversial. In this paper we present the analysis of technical details and immediate and late outcome of a consecutive series of 64 cases of splenic disorders approached by laparoscopy. Between Feb-1993 and April-1997, 64 patients with a wide range of splenic disorders were treated by laparoscopy, and prospectively recorded. Age, body mass index, operative time, number of trocars, perioperative transfusion, spleen weight, conversion rate, mode of spleen retrieval (bag or accessory incision), postoperative analgesia, stay and morbidity were analyzed. Late failures after LS were reevaluated with 99mTc-heat damaged red blood cells scintigraphy and CT. LS was performed in 61 patients, and two cases with splenic cyst and one splenic artery aneurysm received a laparoscopic partial cystectomy and aneurysmectomy. LS was performed through an anterior approach in 12 patients and laterally in 49. Conversion rate was 6.5%. Accessory spleens were found in 7 patients (7/61, 11.5%). Morbidity was 16%. There was no correlation between the weight of the spleen, platelet count or obesity with operative time. A lateral approach was associated with a decrease in operative time (p < 0.002), postoperative stay (p < 0.001), transfusion (p < 0.04) and number of trocars (p < 0.001). Operative time was significantly longer in large spleens (> 1000 gr) (p < 0.001). However, there were no differences in transfusion rate, stay, morbidity or conversion rate. After a follow up of 12 m, 10 patients revealed a low platelet count. Scintigraphy showed residual splenic tissue in 3 (ITP). A wide range of splenic disorders can be treated by laparoscopy, including enlarged spleens. This technique should be continually audited, but initial results reflect the approach's safety and advantages provided that great technical care is taken and an exhaustive search for accessory spleens is conducted. PMID- 9419310 TI - Laparoscopically assisted massive splenectomy. A preliminary report of the technique of early hilar devascularization. AB - Laparoscopic splenectomy has been safely performed for small spleens, but technical limitations have prevented massive splenectomy. We describe a technique of early hilar devascularization to enable massive splenectomy in three patients over the age of 80 years. Massive splenectomy was performed with minimal blood loss and minor morbidity. Early laparoscopic control of the splenic artery and vein will enable the safe removal of the massive spleen, without major laparotomy. Morbidity of splenectomy may be reduced by laparoscopy. PMID- 9419311 TI - Laparoscopic anatomical hepatic resection. Report of four left lobectomies for solid tumors. AB - Four patients underwent a laparoscopic left hepatic resection for solid tumor, two for metastasis from colonic cancer, and two for focal nodular hyperplasia (final diagnosis). The procedure was performed according to the rules of conventional hepatic surgery and cancer surgery. No blood transfusion was necessary. No surgical complication occurred. In malignant disease, laparoscopy allows a good staging and the performance of a real no-touch technique; the specimen is removed in a plastic bag without contact to the abdominal wall. In symptomatic benign disease the esthetic benefit of the laparoscopic approach is real. In asymptomatic benign disease, laparoscopy could allow large biopsies in the case of uncertain diagnosis or dangerous resection. It allows safe resections in the case of small, well-located tumors. This approach requires sophisticated material and extensive experience in both laparoscopy and hepatobiliary surgery. PMID- 9419312 TI - Local epinephrine facilitates laparoscopic Heller myotomy. AB - Incomplete myotomy and mucosal perforation are the most common technical complications of laparoscopic esophageal myotomy. The muscle layers of the lower esophagus are infiltrated with a 1:100,000 epinephrine solution using a thin needle. Gentle pressure is applied with a peanut sponge to diminish the edema produced by the injections. The longitudinal fibers are separated with a dissector and the semicircular fibers are lifted from the submucosa with a dissector or a hook. The muscle transection is done simply by tearing the fibers or cutting them with scissors. No coagulation is required. Infiltration and topical application of epinephrine solution allowed the performance of 22 laparoscopic esophageal myotomies with excellent visualization, complete muscle division, and without any esophageal or gastric perforation. Injection and topical application of epinephrine solution to the area of the esophagus and stomach which will be subjected to myotomy greatly facilitates the procedure and helps to avoid complications. PMID- 9419314 TI - The age of the universe, dark matter, and structure formation. PMID- 9419313 TI - An inexpensive laparoscopic specimen retrieval bag. PMID- 9419315 TI - Measuring cosmological parameters. AB - In this review, the status of measurements of the matter density (Omegam), the vacuum energy density or cosmological constant (OmegaLambda), the Hubble constant (H0), and the ages of the oldest measured objects (t0) are summarized. Three independent types of methods for measuring the Hubble constant are considered: the measurement of time delays in multiply imaged quasars, the Sunyaev-Zel'dovich effect in clusters, and Cepheid-based extragalactic distances. Many recent independent dynamical measurements are yielding a low value for the matter density (Omegam approximately 0.2-0.3). A wide range of Hubble constant measurements appear to be converging in the range of 60-80 km/sec per megaparsec. Areas where future improvements are likely to be made soon are highlighted-in particular, measurements of anisotropies in the cosmic microwave background. Particular attention is paid to sources of systematic error and the assumptions that underlie many of the measurement methods. PMID- 9419316 TI - Globular clusters, Hipparcos, and the age of the galaxy. AB - We discuss the impact of the results from the recent Hipparcos astrometric satellite on distance estimates of galactic globular clusters. Recalibrating the clusters not only implies a relatively small change in the distance to the Large Magellanic Cloud, and hence a rescaling of several estimates of the Hubble constant, but also leads to significantly younger cluster ages. Although the data are not yet conclusive, the results so far point to a likely resolution of the apparent paradox of a universe younger than its constituents, without requiring significant modifications to simple cosmological models. PMID- 9419317 TI - Globular cluster ages. AB - We review two new methods to determine the age of globular clusters (GCs). These two methods are more accurate than the classical isochrone fitting technique. The first method is based on the morphology of the horizontal branch and is independent of the distance modulus of the globular cluster. The second method uses a careful binning of the stellar luminosity function and determines simultaneously the distance and age of the GC. We find that the oldest galactic GCs have an age of 13.5 +/- 2 gigayears (Gyr). The absolute minimum age for the oldest GCs is 10.5 Gyr (with 99% confidence) and the maximum 16.0 Gyr (with 99% confidence). Therefore, an Einstein-De Sitter Universe (Omega = 1) is not totally ruled out if the Hubble constant is about 65 +/- 10 Km s-1 Mpc-1. PMID- 9419318 TI - The age of the universe from nuclear chronometers. AB - An overview is presented of the current situation regarding radioactive dating of the matter of which our Galaxy is comprised. A firm lower bound on the age from nuclear chronometers of approximately 9-10 Gyr is entirely consistent with age determinations from globular clusters and white dwarf cooling histories. The reasonable assumption of an approximately uniform nucleosynthesis rate yields an age for the Galaxy of 12.8 +/- 3 Gyr, which again is consistent with current determinations from other methods. PMID- 9419319 TI - Galaxies and large scale structure at high redshifts. AB - It is now straightforward to assemble large samples of very high redshift (z approximately 3) field galaxies selected by their pronounced spectral discontinuity at the rest frame Lyman limit of hydrogen (at 912 A). This makes possible both statistical analyses of the properties of the galaxies and the first direct glimpse of the progression of the growth of their large-scale distribution at such an early epoch. Here I present a summary of the progress made in these areas to date and some preliminary results of and future plans for a targeted redshift survey at z = 2.7-3.4. Also discussed is how the same discovery method may be used to obtain a "census" of star formation in the high redshift Universe, and the current implications for the history of galaxy formation as a function of cosmic epoch. PMID- 9419320 TI - The microwave background anisotropies: observations. AB - Most cosmologists now believe that we live in an evolving universe that has been expanding and cooling since its origin about 15 billion years ago. Strong evidence for this standard cosmological model comes from studies of the cosmic microwave background radiation (CMBR), the remnant heat from the initial fireball. The CMBR spectrum is blackbody, as predicted from the hot Big Bang model before the discovery of the remnant radiation in 1964. In 1992 the cosmic background explorer (COBE) satellite finally detected the anisotropy of the radiation-fingerprints left by tiny temperature fluctuations in the initial bang. Careful design of the COBE satellite, and a bit of luck, allowed the 30 microK fluctuations in the CMBR temperature (2.73 K) to be pulled out of instrument noise and spurious foreground emissions. Further advances in detector technology and experiment design are allowing current CMBR experiments to search for predicted features in the anisotropy power spectrum at angular scales of 1 degrees and smaller. If they exist, these features were formed at an important epoch in the evolution of the universe--the decoupling of matter and radiation at a temperature of about 4,000 K and a time about 300,000 years after the bang. CMBR anisotropy measurements probe directly some detailed physics of the early universe. Also, parameters of the cosmological model can be measured because the anisotropy power spectrum depends on constituent densities and the horizon scale at a known cosmological epoch. As sophisticated experiments on the ground and on balloons pursue these measurements, two CMBR anisotropy satellite missions are being prepared for launch early in the next century. PMID- 9419321 TI - Cosmic microwave background theory. AB - A long-standing goal of theorists has been to constrain cosmological parameters that define the structure formation theory from cosmic microwave background (CMB) anisotropy experiments and large-scale structure (LSS) observations. The status and future promise of this enterprise is described. Current band-powers in -space are consistent with a DeltaT flat in frequency and broadly follow inflation-based expectations. That the levels are approximately (10(-5))2 provides strong support for the gravitational instability theory, while the Far Infrared Absolute Spectrophotometer (FIRAS) constraints on energy injection rule out cosmic explosions as a dominant source of LSS. Band-powers at 100 suggest that the universe could not have re-ionized too early. To get the LSS of Cosmic Background Explorer (COBE)-normalized fluctuations right provides encouraging support that the initial fluctuation spectrum was not far off the scale invariant form that inflation models prefer: e.g., for tilted Lambda cold dark matter sequences of fixed 13-Gyr age (with the Hubble constant H0 marginalized), ns = 1.17 +/- 0.3 for Differential Microwave Radiometer (DMR) only; 1.15 +/- 0.08 for DMR plus the SK95 experiment; 1.00 +/- 0.04 for DMR plus all smaller angle experiments; 1.00 +/- 0.05 when LSS constraints are included as well. The CMB alone currently gives weak constraints on Lambda and moderate constraints on Omegatot, but theoretical forecasts of future long duration balloon and satellite experiments are shown which predict percent-level accuracy among a large fraction of the 10+ parameters characterizing the cosmic structure formation theory, at least if it is an inflation variant. PMID- 9419322 TI - Primordial nucleosynthesis. AB - With the advent of the new extragalactic deuterium observations, Big Bang nucleosynthesis (BBN) is on the verge of undergoing a transformation. In the past, the emphasis has been on demonstrating the concordance of the BBN model with the abundances of the light isotopes extrapolated back to their primordial values by using stellar and galactic evolution theories. As a direct measure of primordial deuterium is converged upon, the nature of the field will shift to using the much more precise primordial D/H to constrain the more flexible stellar and galactic evolution models (although the question of potential systematic error in 4He abundance determinations remains open). The remarkable success of the theory to date in establishing the concordance has led to the very robust conclusion of BBN regarding the baryon density. This robustness remains even through major model variations such as an assumed first-order quark-hadron phase transition. The BBN constraints on the cosmological baryon density are reviewed and demonstrate that the bulk of the baryons are dark and also that the bulk of the matter in the universe is nonbaryonic. Comparison of baryonic density arguments from Lyman-alpha clouds, x-ray gas in clusters, and the microwave anisotropy are made. PMID- 9419323 TI - The universe at z > 5: when and how did the "dark age" end? AB - This paper considers how the first subgalactic structures produced the UV radiation that ionized the intergalactic medium before z = 5 and the "feedback" effects of the UV radiation on structure formation. The first "pregalaxies" may eventually be detectable by their direct UV emission, with characteristic spectral features at Lyman alpha; high-z supernovae may also be detectable. Other probes of the intergalactic medium beyond z = 5, and of the epochs of reheating and reionization, are discussed, along with possible links between the diffusion of pregalactic metals and the origin of magnetic fields. PMID- 9419324 TI - Particle components of dark matter. AB - Particle candidates for astrophysical dark matter are reviewed, with particular emphasis on the lightest supersymmetric particle and the axion. The former is now constrained by accelerator experiments to have a mass above about 40 GeV, and ongoing searches at accelerators, in space, and in underground experiments have a good chance to detect it. A reevaluation of the constraint on the axion from supernova 1987a leaves open an interesting window where it may be detected if it constitutes the galactic halo. PMID- 9419325 TI - Direct searches for dark matter: recent results. AB - There is abundant evidence for large amounts of unseen matter in the universe. This dark matter, by its very nature, couples feebly to ordinary matter and is correspondingly difficult to detect. Nonetheless, several experiments are now underway with the sensitivity required to detect directly galactic halo dark matter through their interactions with matter and radiation. These experiments divide into two broad classes: searches for weakly interacting massive particles (WIMPs) and searches for axions. There exists a very strong theoretical bias for supposing that supersymmetry (SUSY) is a correct description of nature. WIMPs are predicted by this SUSY theory and have the required properties to be dark matter. These WIMPs are detected from the byproducts of their occasional recoil against nucleons. There are efforts around the world to detect these rare recoils. The WIMP part of this overview focuses on the cryogenic dark matter search (CDMS) underway in California. Axions, another favored dark matter candidate, are predicted to arise from a minimal extension of the standard model that explains the absence of the expected large CP violating effects in strong interactions. Axions can, in the presence of a large magnetic field, turn into microwave photons. It is the slight excess of photons above noise that signals the axion. Axion searches are underway in California and Japan. The axion part of this overview focuses on the California effort. Brevity does not allow me to discuss other WIMP and axion searches, likewise for accelerator and satellite based searches; I apologize for their omission. PMID- 9419326 TI - Galaxy formation. AB - It is argued that within the standard Big Bang cosmological model the bulk of the mass of the luminous parts of the large galaxies likely had been assembled by redshift z approximately 10. Galaxy assembly this early would be difficult to fit in the widely discussed adiabatic cold dark matter model for structure formation, but it could agree with an isocurvature version in which the cold dark matter is the remnant of a massive scalar field frozen (or squeezed) from quantum fluctuations during inflation. The squeezed field fluctuations would be Gaussian with zero mean, and the distribution of the field mass therefore would be the square of a random Gaussian process. This offers a possibly interesting new direction for the numerical exploration of models for cosmic structure formation. PMID- 9419327 TI - X-ray emission from clusters and groups of galaxies. AB - Recent major advances in x-ray imaging and spectroscopy of clusters have allowed the determination of their mass and mass profile out to approximately 1/2 the virial radius. In rich clusters, most of the baryonic mass is in the gas phase, and the ratio of mass in gas/stars varies by a factor of 2-4. The baryonic fractions vary by a factor of approximately 3 from cluster to cluster and almost always exceed 0.09 h50-[3/2] and thus are in fundamental conflict with the assumption of Omega = 1 and the results of big bang nucleosynthesis. The derived Fe abundances are 0.2-0.45 solar, and the abundances of O and Si for low redshift systems are 0.6-1.0 solar. This distribution is consistent with an origin in pure type II supernova. The amount of light and energy produced by these supernovae is very large, indicating their importance in influencing the formation of clusters and galaxies. The lack of evolution of Fe to a redshift of z approximately 0.4 argues for very early enrichment of the cluster gas. Groups show a wide range of abundances, 0.1-0.5 solar. The results of an x-ray survey indicate that the contribution of groups to the mass density of the universe is likely to be larger than 0.1 h50-2. Many of the very poor groups have large x-ray halos and are filled with small galaxies whose velocity dispersion is a good match to the x-ray temperatures. PMID- 9419328 TI - Cosmic velocity fields and their interpretation. AB - We review the current status of our knowledge of cosmic velocity fields, on both small and large scales. A new statistic is described that characterizes the incoherent, thermal component of the velocity field on scales less than 2h-1 Mpc (h is H0/100 km.s-1.Mpc-1, where H0 is the Hubble constant and 1 Mpc = 3.09 x 10(22) m) and smaller. The derived velocity is found to be quite stable across different catalogs and is of remarkably low amplitude, consistent with an effective Omega approximately 0.15 on this scale. We advocate the use of this statistic as a standard diagnostic of the small-scale kinetic energy of the galaxy distribution. The analysis of large-scale flows probes the velocity field on scales of 10-60 h-1 Mpc and should be adequately described by linear perturbation theory. Recent work has focused on the comparison of gravity or density fields derived from whole-sky redshift surveys of galaxies [e.g., the Infrared Astronomical Satellite (IRAS)] with velocity fields derived from a variety of sources. All the algorithms that directly compare the gravity and velocity fields suggest low values of the density parameter, while the POTENT analysis, using the same data but comparing the derived IRAS galaxy density field with the Mark-III derived matter density field, leads to much higher estimates of the inferred density. Since the IRAS and Mark-III fields are not fully consistent with each other, the present discrepancies might result from the very different weighting applied to the data in the competing methods. PMID- 9419329 TI - Measuring the topology of the universe. AB - Observations of microwave background fluctuations can yield information not only about the geometry of the universe but potentially about the topology of the universe. If the universe is negatively curved, then the characteristic scale for the topology of the universe is the curvature radius. Thus, if we are seeing the effects of the geometry of the universe, we can hope to soon see signatures of the topology of the universe. The cleanest signature of the topology of the universe is written on the microwave sky: There should be thousands of pairs of matched circles. These circles can be used to determine the precise topology and volume of the universe. Because we see hundreds of slices through the fundamental domain of the universe, we can use the microwave observations to reconstruct the initial conditions of the entire universe on the scale of a few megaparsecs. PMID- 9419330 TI - Controlling cell cycle and cell fate: common strategies in prokaryotes and eukaryotes. PMID- 9419331 TI - Relationship between transcription and initiation of meiotic recombination: toward chromatin accessibility. PMID- 9419333 TI - Plant science in lac: A continuation of using tools from Escherichia coli in studying gene function in heterologous systems. PMID- 9419334 TI - Limit, logic, and computation. AB - We introduce "ultrafilter limits" into the classical Turing model of computation and develop a paradigm for interpreting the problem of distinguishing the class P from NP as a logical problem of decidability. We use P(NP) to denote decision problems which can be solved on a (nondeterministic) Turing machine in polynomial time. The concept is that in an appropriate limit it may be possible to prove that problems in P are still decidable, so a problem whose limit is undecidable would be established as lying outside of P. PMID- 9419332 TI - Telomerase activity, cell proliferation, and cancer. PMID- 9419335 TI - P/NP, and the quantum field computer. AB - The central problem in computer science is the conjecture that two complexity classes, P (polynomial time) and NP (nondeterministic polynomial time-roughly those decision problems for which a proposed solution can be checked in polynomial time), are distinct in the standard Turing model of computation: P not equal NP. As a generality, we propose that each physical theory supports computational models whose power is limited by the physical theory. It is well known that classical physics supports a multitude of implementation of the Turing machine. Non-Abelian topological quantum field theories exhibit the mathematical features necessary to support a model capable of solving all #P problems, a computationally intractable class, in polynomial time. Specifically, Witten [Witten, E. (1989) Commun. Math. Phys. 121, 351-391] has identified expectation values in a certain SU(2)-field theory with values of the Jones polynomial [Jones, V. (1985) Bull. Am. Math. Soc. 12, 103-111] that are #P-hard [Jaeger, F., Vertigen, D. & Welsh, D. (1990) Math. Proc. Comb. Philos. Soc. 108, 35-53]. This suggests that some physical system whose effective Lagrangian contains a non Abelian topological term might be manipulated to serve as an analog computer capable of solving NP or even #P-hard problems in polynomial time. Defining such a system and addressing the accuracy issues inherent in preparation and measurement is a major unsolved problem. PMID- 9419336 TI - Supercoiling affects the accessibility of glutathione to DNA-bound molecules: positive supercoiling inhibits calicheamicin-induced DNA damage. AB - DNA superhelical tension, an important feature of genomic organization, is known to affect the interactions of intercalating molecules with DNA. However, the effect of torsional tension on nonintercalative DNA-binding chemicals has received less attention. We demonstrate here that the enediyne calicheamicin gamma1I, a strand-breaking agent specific to the minor groove, causes approximately 50% more damage in negatively supercoiled plasmid DNA than in DNA with positive superhelicity. Furthermore, we show that the decrease in damage in positively supercoiled DNA is controlled at the level of thiol activation of the drug. Our results suggest that supercoiling may affect both the activity of nonintercalating genotoxins in vivo and the accessibility of glutathione and other small physiologic molecules to DNA-bound chemicals or reactions occurring in the grooves of DNA. PMID- 9419337 TI - 5' end maturation and RNA editing have to precede tRNA 3' processing in plant mitochondria. AB - We report the characterization and partial purification of potato mitochondrial RNase Z, an endonuclease that generates mature tRNA 3' ends. The enzyme consists of one (or more) protein(s) without RNA subunits. Products of the processing reaction are tRNA molecules with 3' terminal hydroxyl groups and 3' trailers with 5' terminal phosphates. The main processing sites are located immediately 3' to the discriminator and one nucleotide further downstream. This endonucleolytic processing at and close to the tRNA 3' end in potato mitochondria suggests a higher similarity to the eukaryotic than to the prokaryotic tRNA 3' processing pathway. Partial purification and separation of RNase Z from the 5' processing activity RNase P allowed us to determine biochemical characteristics of the enzyme. The activity is stable over broad pH and temperature ranges, with peak activity at pH 8 and 30 degrees C. Optimal concentrations for MgCl2 and KCl are 5 mM and 30 mM, respectively. The potato mitochondrial RNase Z accepts only tRNA precursors with mature 5' ends. The precursor for tRNAPhe requires RNA editing for efficient processing by RNase Z. PMID- 9419338 TI - Twenty proteins containing a C-terminal SOCS box form five structural classes. AB - The four members of the recently identified suppressor of cytokines signaling family (SOCS-1, SOCS-2, SOCS-3, and CIS, where CIS is cytokine-inducible SH2 containing protein) appear, by various means, to negatively regulate cytokine signal transduction. Structurally, the SOCS proteins are composed of an N terminal region of variable length and amino acid composition, a central SH2 domain, and a previously unrecognized C-terminal motif that we have called the SOCS box. By using the SOCS box amino acid sequence consensus, we have searched DNA databases and have identified a further 16 proteins that contain this motif. These proteins fall into five classes based on the protein motifs found N terminal of the SOCS box. In addition to four new SOCS proteins (SOCS-4 to SOCS 7) containing an SH2 domain and a SOCS box, we describe three new families of proteins that contain either WD-40 repeats (WSB-1 and -2), SPRY domains (SSB-1 to -3) or ankyrin repeats (ASB-1 to -3) N-terminal of the SOCS box. In addition, we show that a class of small GTPases also contains a SOCS box. The expression of representative members of each class of proteins differs markedly, as does the regulation of expression by cytokines. The function of the WSB, SSB, and ASB protein families remains to be determined. PMID- 9419339 TI - Negative control of bacterial DNA replication by a cell cycle regulatory protein that binds at the chromosome origin. AB - Caulobacter crescentus divides asymmetrically generating two distinct cell types at each cell division: a stalked cell competent for DNA replication, and a swarmer cell that is unable to initiate DNA replication until it differentiates into a stalked cell later in the cell cycle. The CtrA protein, a member of the response regulator family of the two-component signal transduction system, controls multiple cell cycle processes in Caulobacter and is present in swarmer cells but absent from stalked cells. We report that CtrA binds five sites within the chromosome replication origin in vitro. These sites overlap an essential DnaA box and a promoter in the origin that is essential for replication initiation. Analysis of mutant alleles of ctrA and point mutations in one of the CtrA binding sites in the origin demonstrate that CtrA represses replication in vivo. CtrA mediated repression at the origin thus restricts replication to the stalked cell type. Thus, the direct coupling of chromosome replication with the cell cycle is mediated by the ubiquitous two-component signaling proteins. PMID- 9419340 TI - Nitric oxide and thiol redox regulation of Janus kinase activity. AB - The activation of Janus kinases (JAKs) is crucial for propagation of the proliferative response initiated by many cytokines. The proliferation of various cell lines, particularly those of hematopoietic origin, is also modulated by mediators of oxidative stress such as nitric oxide and thiol redox reagents. Herein we demonstrate that nitric oxide and other thiol oxidants can inhibit the autokinase activity of rat JAK2 in vitro, presumably through oxidation of crucial dithiols to disulfides within JAK2. The reduced form of JAK2 is the most active form, and the oxidized JAK2 form is inactive. Nitric oxide pretreatment of quiescent Ba/F3 cells also inhibits the interleukin 3-triggered in vivo activation of JAK2, a phenomenon that correlates with inhibited proliferation. Furthermore, we observed that the autokinase activity of JAK3 responds in a similar fashion to thiol redox reagents in vitro and to nitric oxide donors in vivo. We suggest that the thiol redox regulation of JAKs may partially explain the generally immunosuppressive effects of nitric oxide and of other thiol oxidants. PMID- 9419341 TI - Drosophila NURF-55, a WD repeat protein involved in histone metabolism. AB - The Drosophila nucleosome remodeling factor (NURF) is a protein complex of four distinct subunits that assists transcription factor-mediated chromatin remodeling. One NURF subunit, ISWI, is related to the transcriptional regulators Drosophila brahma and yeast SWI2/SNF2. We have determined peptide sequences and isolated cDNA clones for a second NURF component (the 55-kDa subunit). Immunological studies show that p55 is an integral subunit of NURF and is generally associated with polytene chromosomes. The predicted sequence of p55 reveals a WD repeat protein that is identical with the 55-kDa subunit of the Drosophila chromatin assembly factor (CAF-1). Given that WD repeat proteins related to p55 are associated with histone deacetylase and histone acetyltransferase, our findings suggest that p55 and its homologs may function as a common platform for the assembly of protein complexes involved in chromatin metabolism. PMID- 9419342 TI - The distal ectodomain of angiotensin-converting enzyme regulates its cleavage secretion from the cell surface. AB - Angiotensin-converting enzyme (ACE) is a type I ectoprotein that is cleaved off the cell surface by a plasma membrane-bound metalloprotease. However, CD4, another type I ectoprotein does not undergo such cleavage-secretion. In this study, we investigated the structural determinants of the ACE protein that regulate the cleavage-secretion process. Substitution and deletion mutations revealed that the cytoplasmic domain, the transmembrane domain, and the juxtamembrane region encompassing the major and the minor cleavage sites of ACE do not regulate its cleavage. Moreover, a chimeric protein containing the distal extracellular domain of CD4 and the juxtamembrane, transmembrane, and the cytoplasmic domains of ACE, although transported to the cell surface, was not cleavage-secreted. In contrast, the distal extracellular domain of ACE was shown to be the important determinant: a protein containing the distal extracellular domain of ACE and the juxtamembrane, transmembrane, and cytoplasmic domain of CD4 was efficiently cleaved off the cell surface. The chimeric protein was cleaved within the CD4 sequence and the responsible enzymatic activity was inhibited by Compound 3, a relatively specific inhibitor of the ACE secretase activity. These results demonstrate that, in a chimeric protein, the distal extracellular domain of a cleavable protein, such as ACE, can induce a proteolytic cleavage within the juxtamembrane domain of an uncleaved protein such as CD4. PMID- 9419344 TI - Sphingoid base 1-phosphate phosphatase: a key regulator of sphingolipid metabolism and stress response. AB - The sphingolipid metabolites ceramide and sphingosine-1-phosphate are second messengers with opposing roles in mammalian cell growth arrest and survival; their relative cellular level has been proposed to be a rheostat that determines the fate of cells. This report demonstrates that this rheostat is an evolutionarily conserved stress-regulatory mechanism that influences growth and survival of yeast. Although the role of sphingosine-1-phosphate in yeast was not previously examined, accumulation of ceramide has been shown to induce G1 arrest and cell death. We now have identified a gene in Saccharomyces cerevisiae, LBP1, that regulates the levels of phosphorylated sphingoid bases and ceramide. LBP1 was cloned from a yeast mutant that accumulated phosphorylated long-chain sphingoid bases and diverted sphingoid base intermediates from sphingolipid pathways to glycerophospholipid biosynthesis. LBP1 and its homolog, LBP2, encode very hydrophobic proteins that contain a novel-conserved sequence motif for lipid phosphatases, and both have long-chain sphingoid base phosphate phosphatase activity. In vitro characterization of Lbp1p shows that this phosphatase is Mg2+ independent with high specificity for phosphorylated long-chain bases, phytosphingosine and sphingosine. The deletion of LBP1 results in the accumulation of phosphorylated long-chain sphingoid bases and reduced ceramide levels. Moreover, deletion of LBP1 and LBP2 results in dramatically enhanced survival upon severe heat shock. Thus, these phosphatases play a previously unappreciated role in regulating ceramide and phosphorylated sphingoid base levels in yeast, and they modulate stress responses through sphingolipid metabolites in a manner that is reminiscent of their effects on mammalian cells. PMID- 9419343 TI - Characterization of mammalian translocase of inner mitochondrial membrane (Tim44) isolated from diabetic newborn mouse kidney. AB - Mammalian translocase of mitochondrial inner membrane (mTim44) was isolated during representational difference analysis of cDNA from diabetic mouse kidney. Streptozotocin-induced diabetic mouse kidney cDNA was prepared and subtracted by normal mouse kidney cDNA. By using one of the isolated cDNA fragments as a screening probe, full-length cDNA of mTim44 was isolated from lambdaZAP kidney cDNA library. At the nucleotide level, mTim44 did not exhibit significant homology with any known genes; however, at the amino acid level, it had 50% similarity and 29% identity with yeast Tim44. C-terminal FLAG epitope-tagged mTim44 fusion protein was transiently expressed in COS7 cells. By using anti-FLAG epitope M2 monoclonal antibody, mTim44 was found to have its subcellular localization associated with mitochondria. By immunoelectron microscopy, mTim44 was seen in the paracrystalline structures within the mitochondria, as well as in their cristae. Mitochondrial import assay of in vitro translated mTim44 indicated that its precursor product ( approximately 50 kDa) was imported and proteolytically processed to a mature approximately 44-kDa protein, and its translocation was inner membrane potential (DeltaPsi)-dependent. Imported mTim44 was protected from protease digestion in which outer membranes were selectively permeabilized with digitonin. The mature form of mTim44 could be recovered in the supernatant of sonicated mitochondrial membrane fraction treated with 0.1 M Na2CO3, pH 11.5. The data indicate that mTim44 is a mitochondrial inner membrane protein, one of the members of the mammalian TIM complex and up-regulated in hyperglycemic states. PMID- 9419346 TI - Hexamethylene bisacetamide induces programmed cell death (apoptosis) and down regulates BCL-2 expression in human myeloma cells. AB - Multiple myeloma (MM) is a B cell malignancy characterized by the expansion of monoclonal Ig-secreting plasma cells with low proliferative activity. It is postulated that inhibition of physiologic cell death is an underlying factor in the pathophysiology of MM. The development of chemoresistance is a common feature in patients with MM. In the present studies, hexamethylene bisacetamide (HMBA), a hybrid polar compound that is a potent inducer of terminal differentiation of various transformed cells, is shown to inhibit the growth of several human myeloma cell lines (ARP-1, U266, and RPMI 8226), including doxorubicin-resistant RPMI 8226 variants that overexpress the multidrug-resistance gene, MDR-1, and its product, p-glycoprotein. In addition to growth arrest and suppression of clonogenicity, HMBA induces apoptosis both in freshly isolated human myeloma cells and in cell lines, as determined by morphologic alterations, cell cycle distribution and endonucleosomal DNA fragmentation. Further, HMBA decreases BCL-2 protein expression in myeloma cells within 12-48 hr. Overexpression of BCL-2 protein in ARP-1 cells confers resistance to HMBA-induced apoptosis. Taken together, these data suggest that HMBA is a potent inducer of apoptosis in human myeloma cells, which may act through suppressing the anti-apoptotic function of the bcl-2 gene. HMBA, and related hybrid polar compounds, may prove useful in the management of this presently incurable disease. PMID- 9419345 TI - Shortage of mitogen-activated protein kinase is responsible for resistance to AP 1 transactivation and transformation in mouse JB6 cells. AB - The JB6 mouse epidermal cell system, which includes tumor promotion-sensitive (P+) and tumor promotion-resistant (P-) cells, is a well-established and extensively used cell culture model for studying the mechanism of late-stage tumor promotion. Tumor promoters, such as 12-O-tetradecanoylphorbol 13-acetate (TPA) or epidermal growth factor (EGF), induce high levels of activator protein 1 (AP-1) activity and large, tumorigenic, anchorage-independent colonies in soft agar at a high frequency in JB6 P+ cells, but not in JB6 P- cells. We report here a molecular explanation for the defect in the AP-1 activation and promotion resistant phenotype of P- cells. We demonstrate that the lack of AP-1 activation and cell transformation responses to TPA and EGF in P- cells appears attributable to the low level of mitogen-activated protein kinase (MAPK) (extracellular signal regulated protein kinase, Erk) in these cells. TPA and EGF induce transactivation of AP-1 activity in P+ cells but not in P- cells. Nonphosphorylated forms and TPA or EGF-induced phosphorylated forms of Erks (Erk1 and Erk2) in P- cells were much lower than those in P+ cells. Stable transfection of wild-type MAPK (Erk2) into P- cells restored its response to TPA and EGF for both AP-1 activation and cell transformation. These results suggest that the shortage of MAPK (Erk1 and Erk2) appears to be an important contributor to the tumor promotion-resistant phenotype in JB6 cells. PMID- 9419347 TI - Creation of monosomic derivatives of human cultured cell lines. AB - Monosomic mammalian cell lines would be ideal for studying gene dosage effects, including gene imprinting, and for systematic isolation of recessive somatic mutants parallel to the invaluable mutants derived from haploid yeast. But autosomal monosomies are lethal in early development; although monosomies appear in tumors, deriving cell lines from these tumors is difficult and cannot provide several syngenic lines. We have developed a strategy for generating stable monosomic human cells, based on random autosomal integration of the gpt plasmid, partial inhibition of DNA topoisomerase II during mitosis to promote chromatid nondisjunction, and selection against retention of gpt. These are likely to be valuable as a source of otherwise inaccessible mutants. The strategy can also be used to generate partial mammalian monosomies, which are desirable as a source of information on recessive genes and gene imprinting. PMID- 9419348 TI - Regulation of interleukin 4-mediated signaling by naturally occurring dominant negative and attenuated forms of human Stat6. AB - Interleukin (IL)-4-mediated nuclear signaling by Stat6 has been implicated in lymphoid cell proliferation and the transcriptional activation of genes encoding major histocompatability complex (MHC) class II molecules and Fc receptors. To investigate IL-4-mediated transcriptional events, we cloned two naturally occurring human Stat6 isoforms, Stat6b and Stat6c, that encoded an NH2-terminal truncation or an SH2 domain deletion, respectively. Stat6 variant mRNAs were differentially expressed in many human tissues. To elucidate the biologic role of each isoform, we examined the consequences of overexpression in IL-4-responsive FDC-P2 cells. Stat6 and Stat6b (to a lesser extent) enhanced DNA synthesis, up regulated endogenous MHC class II and Fcgamma receptors, and became tyrosine phosphorylated in response to IL-4 stimulation. In contrast, Stat6c, which lacks functionally critical SH2 domain residues, unexpectedly inhibited IL-4-mediated mitogenesis and cell surface antigen expression and was not tyrosine phosphorylated. Although Stat6c only modestly diminished endogenous Stat6 tyrosine phosphorylation, it abolished endogenous Stat6 FcgammaRI and Iepsilon DNA binding activity and FcgammaRI-luciferase reporter transcriptional activation. Our results indicate that the molecular mechanism of inhibition by Stat6c was due to suppression of endogenous Stat6 dimer formation. Thus, Stat6b and Stat6c are naturally occurring attenuated and dominant negative Stat6 variants, respectively, that affect IL-4-mediated biologic responses through differential transcriptional regulation. PMID- 9419349 TI - Intestinal trefoil factor induces inactivation of extracellular signal-regulated protein kinase in intestinal epithelial cells. AB - Intestinal trefoil factor (ITF), a small, compact protease-resistant peptide, is abundantly expressed in goblet cells of large and small intestine. Although several biological activities of ITF have been identified, including promotion of wound healing, stimulation of epithelial cell migration, and protection of intestinal epithelial barrier, little is known about signaling events through which ITF mediates its physiological function. In this study, the effects of exogenous ITF on mitogen-activated protein kinase (MAPK) signaling cascades were examined in IEC-6 cells, a nontransformed intestinal epithelial cell line that does not express endogenous trefoil peptides. Stimulation with ITF resulted in rapid decrease in extracellular signal-related protein kinase (ERK) activity and concomitant reduced ERK tyrosine phosphorylation. ITF also decreased activation of ERK activity induced by either transforming growth factor-alpha, which links extracellular stimuli to the Ras/Raf/MEK/ERK pathway via the epidermal growth factor receptor, or phorbol 12-myristate 13-acetate, which activates Raf through protein kinase C. ITF-induced inhibition of ERK activity was blocked by an inhibitor of tyrosine and dual-specific phosphatases, sodium orthovanadate. In summary, ITF leads to inhibition of ERK and the MAPK pathway through activation of tyrosine or dual-specific phosphatase. PMID- 9419350 TI - Two distinct populations of synaptic-like vesicles from rat brain. AB - In nonneuronal cells, several plasma membrane proteins such as exofacial enzymes, receptors, and ion channels recycle between their intracellular compartment(s) and the cell surface via an endosomal pathway. In neurons, however, this pathway has not been extensively characterized. In particular, it remains unclear whether or not it is related to the recycling of small synaptic vesicles, the major pathway of membrane traffic in nerve terminals. To approach this problem, we purified and studied a vesicular fraction from rat brain synaptosomes. Two distinct populations of vesicles with different buoyant densities and sedimentation coefficients were detected in this fraction by sucrose gradient centrifugation and Western blot analysis of the individual proteins. Both populations contain proteins that are markers of synaptic vesicles, namely, SV2, synaptotagmin, synaptophysin, secretory carrier membrane proteins (SCAMPs), synaptobrevin, and rab3a. A striking difference between the two populations is the presence of arginine aminopeptidase activity (a previously suggested marker for the regulated endosomal recycling pathway) exclusively in the lighter less dense vesicles. The same two vesicular populations were also detected in the preparation of clathrin-coated vesicles isolated from whole rat brain or purified synaptosomes after removal of their clathrin coats by incubation at pH 8.5. We conclude, therefore, that both types of vesicles recycle in synaptosomes via a clathrin-mediated pathway. These data present experimental evidence for biochemical heterogeneity of synaptic-like vesicles in rat brain. PMID- 9419351 TI - Consistent loss of functional transforming growth factor beta receptor expression in murine plasmacytomas. AB - Murine plasmacytomas are tumors of Ig-secreting plasma cells that can be induced in genetically susceptible BALB/c mice. The deregulation of the c-myc protooncogene is a critical oncogenic event in the development of plasmacytomas (PCTs) although it is not sufficient for their malignant transformation. We have demonstrated that PCTs produce active transforming growth factor beta (TGF-beta) in vitro. Because TGF-beta is a potent negative regulator of the proliferation and differentiation of B lymphocytes, we examined its role in plasmacytomagenesis by comparing responsiveness to TGF-beta of nonneoplastic plasma cells and PCTs. The nontransformed plasma cells that accumulate in interleukin 6 transgenic mice undergo accelerated apoptosis upon treatment with TGF-beta, but the 15 PCTs studied, including primary and transplanted tumors as well as established cell lines, were refractory to TGF-beta-mediated growth inhibition and apoptosis. Although PCTs lack functional TGF-beta receptors as demonstrated by chemical crosslinking to radiolabeled TGF-beta1, they nonetheless contain mRNA and protein for both type I and II TGF-beta receptors, suggesting a potential defect in receptor trafficking or processing. The results clearly show the consistent inactivation of TGF-beta receptors in plasmacytoma cells, demonstrating for the first time that interruption of a tumor suppressor pathway contributes to plasmacytomagenesis. PMID- 9419352 TI - Synergistic activation of p53 by inhibition of MDM2 expression and DNA damage. AB - The MDM2 oncogene encodes an inhibitor of the p53 tumor suppressor protein that regulates p53 in a negative feedback loop. MDM2 gene amplification and overexpression occur in several types of tumors and are often associated with poor prognosis. An MDM2 antisense phosphorothioate oligodeoxynucleotide has been identified that effectively inhibits MDM2 expression in tumor cells containing MDM2 gene amplifications. Antisense inhibition of MDM2 is associated with a decrease in MDM2-p53 complex formation, increase in p53-inducible gene expression, increase in p53 transcriptional activity, and apoptosis. Significantly, inhibition of MDM2 expression enhances the activation of p53 by a DNA-damaging cancer chemotherapy agent in a synergistic fashion. Therefore, the MDM2 negative feedback pathway is an important limiting factor in DNA damage induced p53 activation. MDM2 antisense oligonucleotides may be useful as antitumor agents alone or as enhancers of other conventional DNA-damaging drugs. PMID- 9419354 TI - Biodiversity at the edge: a test of the importance of spatial "mass effects" in the Rothamsted Park Grass experiments. AB - The coexistence of many plant species competing for a few resources is one of the central puzzles of community ecology. One explanation is that different species may be competitively superior in different microhabitats. Many species could then coexist within each piece of a mosaic landscape by what has been termed "mass effects," because subpopulations in areas with negative growth rates would be supplemented by propagules from areas with reproductive surpluses. If mass effects are important, plant species diversity should increase near habitat boundaries, especially where habitat differences are moderate. In the first experimental test of this prediction, plants were censused on 54 transects within the long-established Rothamsted Park Grass plots. Very few showed significant declines in species richness with distance from subplot boundaries. Nonetheless, the regression coefficients were negative much more often than expected by chance, suggesting that weak mass effects operated. The effect was strongest where neighboring subplots differed greatly, with no evidence of the predicted decline where differences were extreme. Detailed analyses of transects with apparent mass effects revealed few species that behaved as predicted. This study serves both to provide evidence of the existence of mass effects and to question their importance in the maintenance of local plant diversity in this system. PMID- 9419353 TI - Control of direction of flagellar rotation in bacterial chemotaxis. AB - The motile behavior of the bacterium Escherichia coli depends on the direction of rotation of its flagellar motors. Binding of the phosphorylated signaling molecule CheY to a motor component FliM is known to enhance clockwise rotation. It is difficult to study this interaction in vivo, because the dynamics of phosphorylation of CheY by its kinase CheA and the hydrolysis of CheY (accelerated by CheZ) are not under direct experimental control. Here, we examine instead the interaction with the flagellar motor of a double mutant CheY13DK106YW that is active without phosphorylation. The behavioral assays were carried out on tethered cells lacking CheA and CheZ. The effects of variation in intracellular concentration of the mutant protein were highly nonlinear. However, they can be explained by a thermal isomerization model in which the free energies of clockwise and counterclockwise states depend linearly on the amount of CheY bound. PMID- 9419355 TI - Patterns of variance in stage-structured populations: evolutionary predictions and ecological implications. AB - Variability in population growth rate is thought to have negative consequences for organism fitness. Theory for matrix population models predicts that variance in population growth rate should be the sum of the variance in each matrix entry times the squared sensitivity term for that matrix entry. I analyzed the stage specific demography of 30 field populations from 17 published studies for pattern between the variance of a demographic term and its contribution to population growth. There were no instances in which a matrix entry both was highly variable and had a large effect on population growth rate; instead, correlations between estimates of temporal variance in a term and contribution to population growth (sensitivity or elasticity) were overwhelmingly negative. In addition, survivorship or growth sensitivities or elasticities always exceeded those of fecundity, implying that the former two terms always contributed more to population growth rate. These results suggest that variable life history stages tend to contribute relatively little to population growth rates because natural selection may alter life histories to minimize stages with both high sensitivity and high variation. PMID- 9419356 TI - Relationship between "proto-splice sites" and intron phases: evidence from dicodon analysis. AB - The coding sequence at the boundaries of exons flanking nuclear introns shows some degree of conservation. To the extent that such sequences might be recognized by the splicing machinery, this conservation may be a derived result of evolution for efficient splicing. Alternatively, such conserved sequences might be remnants of proto-splice sites, which might have existed early in eukaryotic genes and served as the targets for the insertion of introns, as has been proposed by the introns-late theory. The distribution of intron phases, the position of the intron within a codon, is biased with an over-representation of phase 0 introns. Could any distribution of proto-splice sites account for today's intron phase distribution? Here, we examine the dicodon usage in six model organisms, based on current sequences in the GenBank database, and predict the phase distribution that would be expected if introns had been inserted into proto splice sites. However, these predictions differ between the various model organisms and disagree with the observed intron phase distributions. Thus, we reject the hypothesis that introns are inserted into hypothetical proto-splice sites. Finally, we analyze the sequences around the splice sites of introns in all six of the species to show that the actual conservation of sequence in exon regions near introns is very small and differs considerably between these species, which is inconsistent with a general proto-splice sites model. PMID- 9419357 TI - Universally conserved translation initiation factors. AB - The process by which translation is initiated has long been considered similar in Bacteria and Eukarya but accomplished by a different unrelated set of factors in the two cases. This not only implies separate evolutionary histories for the two but also implies that at the universal ancestor stage, a translation initiation mechanism either did not exist or was of a different nature than the extant processes. We demonstrate herein that (i) the "analogous" translation initiation factors IF-1 and eIF-1A are actually related in sequence, (ii) the "eukaryotic" translation factor SUI1 is universal in distribution, and (iii) the eukaryotic/archaeal translation factor eIF-5A is homologous to the bacterial translation factor EF-P. Thus, the rudiments of translation initiation would seem to have been present in the universal ancestor stage. However, significant development and refinement subsequently occurred independently on both the bacterial lineage and on the archaeal/eukaryotic line. PMID- 9419358 TI - A mitochondrial-like chaperonin 60 gene in Giardia lamblia: evidence that diplomonads once harbored an endosymbiont related to the progenitor of mitochondria. AB - Diplomonads, parabasalids, as represented by trichomonads, and microsporidia are three protist lineages lacking mitochondria that branch earlier than all other eukaryotes in small subunit rRNA and elongation factor phylogenies. The absence of mitochondria and plastids in these organisms suggested that they diverged before the origin of these organelles. However, recent discoveries of mitochondrial-like heat shock protein 70 and/or chaperonin 60 (cpn60) genes in trichomonads and microsporidia imply that the ancestors of these two groups once harbored mitochondria or their endosymbiotic progenitors. In this report, we describe a mitochondrial-like cpn60 homolog from the diplomonad parasite Giardia lamblia. Northern and Western blots reveal that the expression of cpn60 is independent of cellular stress and, except during excystation, occurs throughout the G. lamblia life cycle. Phylogenetic analyses position the G. lamblia cpn60 in a clade that includes mitochondrial and hydrogenosomal cpn60 proteins. The most parsimonious interpretation of these data is that the cpn60 gene was transferred from the endosymbiotic ancestors of mitochondria to the nucleus early in eukaryotic evolution, before the divergence of the diplomonads and trichomonads from other extant eukaryotic lineages. A more complicated explanation requires that these genes originated from distinct alpha-proteobacterial endosymbioses that formed transiently within these protist lineages. PMID- 9419359 TI - Nuclear ribosomal DNA evidence for a western North American origin of Hawaiian and South American species of Sanicula (Apiaceae). AB - Results from phylogenetic analysis of nuclear rDNA internal transcribed spacer (ITS) sequences from a worldwide sample of Sanicula indicate that Hawaiian sanicles (Sanicula sect. Sandwicenses) constitute a monophyletic group that descended from a western North American ancestor in Sanicula sect. Sanicoria, a paraphyletic assemblage of mostly Californian species. A monophyletic group comprising representatives of all 15 species of S. sect. Sanicoria and the three sampled species of S. sect. Sandwicenses was resolved in all maximally parsimonious trees, rooted with sequences from species of Astrantia and Eryngium. All sequences sampled from eastern North American, European, and Asian species of Sanicula fell outside the ITS clade comprising S. sect. Sanicoria and S. sect. Sandwicenses. A lineage comprising the Hawaiian taxa and three species endemic to coastal or near-coastal habitats in western North America (Sanicula arctopoides, Sanicula arguta, and Sanicula laciniata) is diagnosed by nucleotide substitutions and a 24-bp deletion in ITS2. The hooked fruits in Sanicula lead us to conclude that the ancestor of Hawaiian sanicles arrived from North America by external bird dispersal; similar transport has been hypothesized for the North American tarweed ancestor of the Hawaiian silversword alliance (Asteraceae). Two additional long-distance dispersal events involving members of S. sect. Sanicoria can be concluded from the ITS phylogeny: dispersal of Sanicula crassicaulis and Sanicula graveolens from western North America to southern South America. PMID- 9419361 TI - Assaying genome-wide recombination and centromere functions with Arabidopsis tetrads. AB - During meiosis, crossover events generate new allelic combinations, yet the abundance of these genetic exchanges in individual cells has not been measured previously on a genomic level. To perform a genome-wide analysis of recombination, we monitored the assortment of genetic markers in meiotic tetrads from Arabidopsis. By determining the number and distribution of crossovers in individual meiotic cells, we demonstrated (i) surprisingly precise regulation of crossover number in each meiosis, (ii) considerably reduced recombination along chromosomes carrying ribosomal DNA arrays, and (iii) an inversely proportional relationship between recombination frequencies and chromosome size. This use of tetrad analysis also achieved precise mapping of all five Arabidopsis centromeres, localizing centromere functions in the intact chromosomes of a higher eukaryote. PMID- 9419360 TI - beta-dystrobrevin, a member of the dystrophin-related protein family. AB - The importance of dystrophin and its associated proteins in normal muscle function is now well established. Many of these proteins are expressed in nonmuscle tissues, particularly the brain. Here we describe the characterization of beta-dystrobrevin, a dystrophin-related protein that is abundantly expressed in brain and other tissues, but is not found in muscle. beta-dystrobrevin is encoded by a 2.5-kb alternatively spliced transcript that is found throughout the brain. In common with dystrophin, beta-dystrobrevin is found in neurons of the cortex and hippocampal formation but is not found in the brain microvasculature. In the brain, beta-dystrobrevin coimmunoprecipitates with the dystrophin isoforms Dp71 and Dp140. These data provide evidence that the composition of the dystrophin-associated protein complex in the brain differs from that in muscle. This finding may be relevant to the cognitive dysfunction affecting many patients with Duchenne muscular dystrophy. PMID- 9419362 TI - Marginal fitness contributions of nonessential genes in yeast. AB - Analysis of the complete genome sequence of Saccharomyces cerevisiae confirms and extends earlier evidence that a majority of yeast genes are not essential, at least under laboratory conditions. Many fail to yield a discernible mutant phenotype even when disrupted. Genes not subject to natural selection would accumulate inactivating mutations, so these "cryptic" genes must have functions that are overlooked by the standard methods of yeast genetics. Two explanations seem possible: (i) They have important functions only in environments not yet duplicated in the laboratory and would have conditional phenotypes if tested appropriately. (ii) They make small, but significant, contributions to fitness even under routine growth conditions, but the effects are not large enough to be detected by conventional methods. We have tested the second "marginal benefit" hypothesis by measuring the fitnesses of a random collection of disruption mutants in direct competition with their wild-type progenitor. A substantial majority of mutant strains that lack obvious defects nevertheless are at a significant selective disadvantage just growing on rich medium under normal conditions. This result has important implications for efforts to understand the functions of novel genes revealed by sequencing projects. PMID- 9419363 TI - A chemokine expressed in lymphoid high endothelial venules promotes the adhesion and chemotaxis of naive T lymphocytes. AB - Preferential homing of naive lymphocytes to secondary lymphoid organs is thought to involve the action of chemokines, yet no chemokine has been shown to have either the expression pattern or the activities required to mediate this process. Here we show that a chemokine represented in the EST database, secondary lymphoid tissue chemokine (SLC), is expressed in the high endothelial venules of lymph nodes and Peyer's patches, in the T cell areas of spleen, lymph nodes, and Peyer's patches, and in the lymphatic endothelium of multiple organs. SLC is a highly efficacious chemoattractant for lymphocytes with preferential activity toward naive T cells. Moreover, SLC induces firm adhesion of naive T lymphocytes via beta2 integrin binding to the counter receptor, intercellular adhesion molecule-1, a necessary step for lymphocyte recruitment. SLC is the first chemokine demonstrated to have the characteristics required to mediate homing of lymphocytes to secondary lymphoid organs. In addition, the expression of SLC in lymphatic endothelium suggests that the migration of lymphocytes from tissues into efferent lymphatics may be an active process mediated by this molecule. PMID- 9419364 TI - Heteroclitic proliferative responses and changes in cytokine profile induced by altered peptides: implications for autoimmunity. AB - Productive engagement of T cell receptors (TCRs) by cognate ligand (major histocompatibility complex plus peptide) leads to proliferation, differentiation, and the elaboration of effector functions. Altered peptides generated by single amino acid substitutions in the antigenic peptide have diverse effects on the outcome of the T cell response. We have generated an altered peptide (Q144) from an autoantigenic peptide of myelin proteolipid protein 139-151 by a single amino acid substitution (from tryptophan to glutamine) in the primary TCR contact at position 144 that is capable of inducing CD4(+) T cell responses in H-2(s) mice. By using a Q144-specific T cell clone (Q1.1B6), we see a hierarchy in T cell proliferation and cytokine production with various position 144 substituted peptides and have identified a peptide (L144) that hyperstimulates this T cell clone. In contrast to Q144, L144 induces maximal proliferation at 7 logs lower antigen concentration, induces greater cell death at higher antigen dose, and induces the secretion of cytokines not detected following stimulation with the cognate ligand. This heteroclitic T cell response associated with changes in cytokine profile was observed with several other T cell clones of different specificities. The L144 peptide also induces costimulation independent proliferation and cytokine production from the Q1.1B6 T cell clone. We describe this as a superagonist response. Such responses may have a role in the initiation of autoimmunity by promoting a proinflammatory environment following ligation of a cross-reactive TCR on autoreactive T cells. PMID- 9419365 TI - Human cytolytic and interferon gamma-secreting CD8+ T lymphocytes specific for Mycobacterium tuberculosis. AB - Protective immunity to Mycobacterium tuberculosis is poorly understood, but mounting evidence, at least in animal models, implicates major histocompatibility complex class I-restricted CD8+ T cells as an essential component. By using a highly sensitive assay for single cell interferon gamma release, we screened an array of M. tuberculosis antigen-derived peptides congruent with HLA class I allele-specific motifs. We identified CD8+ T cells specific for epitopes in the early secretory antigenic target 6 during active tuberculosis, after clinical recovery and in healthy contacts. Unrestimulated cells exhibited peptide-specific interferon gamma secretion, whereas lines or clones recognized endogenously processed antigen and showed cytolytic activity. These results provide direct evidence for the involvement of CD8+ cytotoxic T lymphocytes in host defense against M. tuberculosis in humans and support current attempts to generate protective cytotoxic T lymphocyte responses against M. tuberculosis by vaccination. PMID- 9419366 TI - Stimulation of HIV-1-neutralizing antibodies in simian HIV-IIIB-infected macaques. AB - Previously we have discovered a public idiotope, designated 1F7, that is expressed on antibodies against HIV type 1 (HIV-1) in human and nonhuman primates. To test the potential of mouse monoclonal antibody (mAb) 1F7 as a therapeutic anti-clonotypic antibody in HIV-1-infected patients, we used the simian HIV-IIIB macaque infection model, which mimics several immunological and pathological characteristics of HIV-1 infection in humans. Four healthy simian HIV-infected rhesus monkeys (Macaca mulatta) expressing the 1F7 marker on anti gp120 antibodies were selected for this study. Three monkeys of this group were immunized several times with the murine mAb 1F7 i.v., and one monkey received as control an isotype-matched antibody, TEPC183. No serious side effect or allergic reaction was encountered. Blood collected before and during the immunization and over several months afterward were analyzed for neutralizing antibodies. Significant increases in breadth and potency of HIV-1-neutralizing antibody titers to one or more virus strains were detected in all three of the 1F7 immunized monkeys, but not in the control monkey immunized with TEPC183. These results show that an antibody, recognizing a public idiotope associated with anti HIV-1 antibodies can function in chronically infected primates as an anti clonotypic immunogen to boost antibodies that neutralize homologous and heterologous virus strains. This study represents a first step toward the preclinical evaluation of 1F7 as a therapeutic AIDS vaccine. PMID- 9419367 TI - Mutations in the MDR3 gene cause progressive familial intrahepatic cholestasis. AB - Class III multidrug resistance (MDR) P-glycoproteins (P-gp), mdr2 in mice and MDR3 in man, mediate the translocation of phosphatidylcholine across the canalicular membrane of the hepatocyte. Mice with a disrupted mdr2 gene completely lack biliary phospholipid excretion and develop progressive liver disease, characterized histologically by portal inflammation, proliferation of the bile duct epithelium, and fibrosis. This disease phenotype is very similar to a subtype of progressive familial intrahepatic cholestasis, hallmarked by a high serum gamma-glutamyltransferase (gamma-GT) activity. We report immunohistochemistry for MDR3 P-gp, reverse transcription-coupled PCR sequence analysis, and genomic DNA analysis of MDR3 from two progressive familial intrahepatic cholestasis patients with high serum gamma-GT. Canalicular staining for MDR3 P-gp was negative in liver tissue of both patients. Reverse transcription-coupled PCR sequencing of the first patient's sequence demonstrated a homozygous 7-bp deletion, starting at codon 132, which results in a frameshift and introduces a stop codon 29 codons downstream. The second patient is homozygous for a nonsense mutation in codon 957 (C --> T) that introduces a stop codon (TGA). Our results demonstrate that mutations in the human MDR3 gene lead to progressive familial intrahepatic cholestasis with high serum gamma-GT. The histopathological picture in these patients is very similar to that in the corresponding mdr2(-/-) mouse, in which mdr2 P-gp deficiency induces complete absence of phospholipid in bile. PMID- 9419368 TI - DNA oxidation matters: the HPLC-electrochemical detection assay of 8-oxo deoxyguanosine and 8-oxo-guanine. AB - Oxidative DNA damage is important in aging and the degenerative diseases of aging such as cancer. Estimates commonly rely on measurements of 8-oxo-2' deoxyguanosine (oxo8dG), an adduct that occurs in DNA and is also excreted in urine after DNA repair. Here we examine difficulties inherent in the analysis of oxo8dG, identify sources of artifacts, and provide solutions to some of the common methodological problems. A frequent criticism has been that phenol in DNA extraction solutions artificially increases the measured level of oxo8dG. We found that phenol extraction of DNA contributes a real but minor increase in the level of oxo8dG when compared, under equivalent conditions, with a successful nonphenol method. A more significant reduction in the baseline level was achieved with a modification of the recently introduced chaotropic NaI method, reducing our estimate of the level of steady-state oxidative adducts by an order of magnitude to 24,000 adducts per cell in young rats and 66,000 adducts per cell in old rats. Of several alternative methods tested, the use of this chaotropic technique of DNA isolation by using NaI produced the lowest and least variable oxo8dG values. In further studies we show that human urinary 8-oxo-guanine (oxo8Gua) excretion is not affected by the administration of allopurinol, suggesting that, unlike some methylated adducts, oxo8Gua is not derived enzymatically from xanthine oxidase. Lastly, we discuss remaining uncertainties inherent both in steady-state oxo8dG measurements and in estimates of endogenous oxidation ("hit rates") based on urinary excretion of oxo8dG and oxo8Gua. PMID- 9419369 TI - Measurement and modeling of the transient difference between blood and subcutaneous glucose concentrations in the rat after injection of insulin. AB - The kinetics of the fall in subcutaneous fluid glucose concentration in anesthetized rats (n = 7) after intravenous injection of insulin (0.5 units/kg) was studied by using 5 x 10(-4) cm2 active area, <150-sec 10-90% response time, amperometric glucose sensors. The onset of the decline in the subcutaneous glucose concentration was delayed and statistically different (P < 0.001) from that in blood (8.9 +/- 2.1 min vs. 3.3 +/- 0.5 min). Similarly, the rate of drop in glucose concentration between 6 and 20 min after the insulin injection was different for subcutaneous tissue (3.9 +/- 1.3 mg.dl-1. min-1) and blood (6.8 +/- 2.0 mg.dl-1.min-1) (P = 0.003). The hypoglycemic nadir in subcutaneous fluid occurred 24.5 +/- 6.8 min after that in the blood (P < 0.001). A "forward" mass transfer model, predicting the subcutaneous glucose concentration from the blood glucose concentrations and an "inverse" model, predicting the blood glucose concentration from the subcutaneous glucose concentration were derived. By using an algorithm based on the latter, the average discrepancy between the measured blood glucose concentration and that estimated from the subcutaneous measurement through the entire 4-hr experiment was reduced from 22.9% to 11.1% (P = 0.025). The maximum discrepancy during the 40-min period after the injection of insulin was reduced from 84.1% to 29.3% (P = 0.006). PMID- 9419370 TI - Discovery of three genes specifically expressed in human prostate by expressed sequence tag database analysis. AB - A procedure is described to discover genes that are specifically expressed in human prostate. The procedure involves searching the expressed sequence tag (EST) database for genes that have many related EST sequences from human prostate cDNA libraries but none or few from nonprostate human libraries. The selected candidate EST clones were tested by RNA dot blots to examine tissue specificity and by Northern blots to examine the transcript size of the corresponding mRNA. The computer analysis identified 15 promising genes that were previously unidentified. When seven of these were examined in an RNA hybridization experiment, three were found to be prostate specific. The genes identified could be useful in the targeted therapy of prostate cancer. The procedure can easily be applied to discover genes specifically expressed in other organs or tumors. PMID- 9419371 TI - Second gene for gonadotropin-releasing hormone in humans. AB - Gonadotropin-releasing hormone (GnRH) is a decapeptide widely known for its role in regulating reproduction by serving as a signal from the hypothalamus to pituitary gonadotropes. In addition to hypothalamic GnRH (GnRH-I), a second GnRH form (pGln-His-Trp-Ser-His-Gly-Trp-Tyr-Pro-Gly; GnRH-II) with unknown function has been localized to the midbrain of many vertebrates. We show here that a gene encoding GnRH-II is expressed in humans and is located on chromosome 20p13, distinct from the GnRH-I gene that is on 8p21-p11.2. The GnRH-II genomic and mRNA structures parallel those of GnRH-I. However, in contrast to GnRH-I, GnRH-II is expressed at significantly higher levels outside the brain (up to 30x), particularly in the kidney, bone marrow, and prostate. The widespread expression of GnRH-II suggests it may have multiple functions. Molecular phylogenetic analysis shows that this second gene is likely the result of a duplication before the appearance of vertebrates, and predicts the existence of a third GnRH form in humans and other vertebrates. PMID- 9419372 TI - Liver repopulation with xenogenic hepatocytes in B and T cell-deficient mice leads to chronic hepadnavirus infection and clonal growth of hepatocellular carcinoma. AB - To investigate host and viral mechanisms determining hepadnaviral persistence and hepatocarcinogenesis, we developed a mouse model by transplanting woodchuck hepatocytes into the liver of mice that contain the urokinase-type plasminogen activator transgene (uPA) and lack mature B and T lymphocytes due to a recombination activation gene 2 (RAG-2) gene knockout. The woodchuck hepatocytes were transplanted via intrasplenic injection and were found to integrate into the recipient mouse liver cord structure. Normal adult woodchuck hepatocytes proliferated and reconstituted up to 90% of the uPA/RAG-2 mouse liver. uPA/RAG-2 mice containing woodchuck hepatocytes were infectable with woodchuck hepatitis virus (WHV) and showed WHV replication for at least 10 months with titers up to 1 x 10(11) virions per ml in the peripheral blood. WHV-infected hepatocytes from chronic carrier woodchucks also established a persistent infection in uPA/RAG-2 mice after an 8- to 12-week lag period of viremia. Although WHV envelope, core, and X proteins were produced in the uPA/RAG-2 mice, no inflammatory host immune response was observed in the liver of WHV-replicating mice. A first antiviral test demonstrated a greater than four orders of magnitude drop in WHV titer in response to interferon alpha treatment. WHV replication was up-regulated by dexamethasone treatment. Comparison of precancerous lesions in donor woodchucks versus recipient uPA/RAG-2 mice revealed an enrichment of dysplastic precancerous hepatocytes in transplanted mice. Clonal amplification of hepatocytes from a woodchuck with hepatocellular carcinomas was demonstrated by the detection of unique WHV DNA integration patterns in hepatocellular carcinomas that arose in uPA/RAG-2 mice. In the absence of B or T cell-mediated immune responses, WHV establishes a persistent noncytotoxic infection of woodchuck hepatocytes in uPA/RAG-2 chimeric mouse livers. Further studies of the kinetics of hepadnavirus infection and replication in quiescent and proliferating hepatocytes should increase our understanding of hepadnavirus spread and aid in the design of therapies to block or cure persistent infection. PMID- 9419373 TI - Stoichiometric coupling of brain glucose metabolism and glutamatergic neuronal activity. AB - To determine the relationship between cerebral Glc metabolism and glutamatergic neuronal function, we used 13C NMR spectroscopy to measure, simultaneously, the rates of the tricarboxylic acid cycle and Gln synthesis in the rat cortex in vivo. From these measurements, we calculated the rates of oxidative Glc metabolism and glutamate-neurotransmitter cycling between neurons and astrocytes (a quantitative measure of glutamatergic neuronal activity). By measuring the rates of the tricarboxylic acid cycle and Gln synthesis over a range of synaptic activity, we have determined the stoichiometry between oxidative Glc metabolism and glutamate-neurotransmitter cycling in the cortex to be close to 1:1. This finding indicates that the majority of cortical energy production supports functional (synaptic) glutamatergic neuronal activity. Another implication of this result is that brain activation studies, which map cortical oxidative Glc metabolism, provide a quantitative measure of synaptic glutamate release. PMID- 9419374 TI - The hypocretins: hypothalamus-specific peptides with neuroexcitatory activity. AB - We describe a hypothalamus-specific mRNA that encodes preprohypocretin, the putative precursor of a pair of peptides that share substantial amino acid identities with the gut hormone secretin. The hypocretin (Hcrt) protein products are restricted to neuronal cell bodies of the dorsal and lateral hypothalamic areas. The fibers of these neurons are widespread throughout the posterior hypothalamus and project to multiple targets in other areas, including brainstem and thalamus. Hcrt immunoreactivity is associated with large granular vesicles at synapses. One of the Hcrt peptides was excitatory when applied to cultured, synaptically coupled hypothalamic neurons, but not hippocampal neurons. These observations suggest that the hypocretins function within the CNS as neurotransmitters. PMID- 9419375 TI - Null mutation in the rhodopsin kinase gene slows recovery kinetics of rod and cone phototransduction in man. AB - Rhodopsin kinase (RK), a specialized G-protein-coupled receptor kinase expressed in retina, is involved in quenching of light-induced signal transduction in photoreceptors. The role of RK in recovery after photoactivation has been explored in vitro and in vivo experimentally but has not been specifically defined in humans. We investigated the effects on human vision of a mutation in the RK gene causing Oguchi disease, a recessively inherited retinopathy. In vitro experiments demonstrated that the mutation, a deletion of exon 5, abolishes the enzymatic activity of RK and is likely a null. Both a homozygote and heterozygote with this RK mutation had recovery phase abnormalities of rod-isolated photoresponses by electroretinography (ERG); photoactivation was normal. Kinetics of rod bleaching adaptation by psychophysics were dramatically slowed in the homozygote but normal final thresholds were attained. Light adaptation was normal at low backgrounds but became abnormal at higher backgrounds. A slight slowing of cone deactivation kinetics in the homozygote was detected by ERG. Cone bleaching adaptation and background adaptation were normal. In this human in vivo condition without a functional RK and probable lack of phosphorylation and arrestin binding to activated rhodopsin, reduction of photolyzed chromophore and regeneration processes with 11-cis-retinal probably constitute the sole pathway for recovery of rod sensitivity. The role of RK in rods would thus be to accelerate inactivation of activated rhodopsin molecules that in concert with regeneration leads to the normal rate of recovery of sensitivity. Cones may rely mainly on regeneration for the inactivation of photolyzed visual pigment, but RK also contributes to cone recovery. PMID- 9419376 TI - Large-scale temporal gene expression mapping of central nervous system development. AB - We used reverse transcription-coupled PCR to produce a high-resolution temporal map of fluctuations in mRNA expression of 112 genes during rat central nervous system development, focusing on the cervical spinal cord. The data provide a temporal gene expression "fingerprint" of spinal cord development based on major families of inter- and intracellular signaling genes. By using distance matrices for the pair-wise comparison of these 112 temporal gene expression patterns as the basis for a cluster analysis, we found five basic "waves" of expression that characterize distinct phases of development. The results suggest functional relationships among the genes fluctuating in parallel. We found that genes belonging to distinct functional classes and gene families clearly map to particular expression profiles. The concepts and data analysis discussed herein may be useful in objectively identifying coherent patterns and sequences of events in the complex genetic signaling network of development. Functional genomics approaches such as this may have applications in the elucidation of complex developmental and degenerative disorders. PMID- 9419377 TI - Melanopsin: An opsin in melanophores, brain, and eye. AB - We have identified an opsin, melanopsin, in photosensitive dermal melanophores of Xenopus laevis. Its deduced amino acid sequence shares greatest homology with cephalopod opsins. The predicted secondary structure of melanopsin indicates the presence of a long cytoplasmic tail with multiple putative phosphorylation sites, suggesting that this opsin's function may be finely regulated. Melanopsin mRNA is expressed in hypothalamic sites thought to contain deep brain photoreceptors and in the iris, a structure known to be directly photosensitive in amphibians. Melanopsin message is also localized in retinal cells residing in the outermost lamina of the inner nuclear layer where horizontal cells are typically found. Its expression in retinal and nonretinal tissues suggests a role in vision and nonvisual photoreceptive tasks, such as photic control of skin pigmentation, pupillary aperture, and circadian and photoperiodic physiology. PMID- 9419378 TI - Gsalpha-selective G protein antagonists. AB - Suramin acts as a G protein inhibitor because it inhibits the rate-limiting step in activation of the Galpha subunit, i.e., the exchange of GDP for GTP. Here, we have searched for analogues that are selective for Gsalpha. Two compounds have been identified: NF449 (4,4',4",4'"-[carbonyl-bis[imino-5,1,3-benzenetriyl bis (carbonylimino)]]tetrakis-(benzene-1,3-disulfonate) and NF503 (4, 4' [carbonylbis[imino-3,1-phenylene-(2, 5-benzimidazolylene)carbonylimino]]bis benzenesulfonate). These compounds (i) suppress the association rate of guanosine 5'-[gamma-thio]triphosphate ([35S]GTP[gammaS]) binding to Gsalpha-s but not to Gialpha-1, (ii) inhibit stimulation of adenylyl cyclase activity in S49 cyc- membranes (deficient in endogenous Gsalpha) by exogenously added Gsalpha-s, and (iii) block the coupling of beta-adrenergic receptors to Gs with half-maximum effects in the low micromolar range. In contrast to suramin, which is not selective, NF503 and NF449 disrupt the interaction of the A1-adenosine receptor with its cognate G proteins (Gi/Go) at concentrations that are >30-fold higher than those required for uncoupling of beta-adrenergic receptor/Gs tandems; similarly, the angiotensin II type-1 receptor (a prototypical Gq-coupled receptor) is barely affected by the compounds. Thus, NF503 and NF449 fulfill essential criteria for Gsalpha-selective antagonists. The observations demonstrate the feasibility of subtype-selective G protein inhibition. PMID- 9419379 TI - Controlling signaling with a specifically designed Gi-coupled receptor. AB - We are developing a system to control G protein signaling in vivo to regulate a broad range of physiologic responses. Our system utilizes G protein-coupled peptide receptors engineered to respond exclusively to synthetic small molecule ligands and not to their natural ligand(s). These engineered receptors are designated RASSLs (receptor activated solely by a synthetic ligand). We have made two prototype RASSLs that are based on the human kappa opioid receptor. Small molecule drugs that activate the kappa receptor are nonaddictive and safe to administer in vivo. Binding and signaling assays reveal 200-2000-fold reductions in the ability of our RASSLs to bind or be activated by dynorphin, an endogenous peptide ligand of the kappa opioid receptor. In a high-throughput signaling assay, these prototype RASSLs expressed in Chinese hamster ovary K1 cells showed little or no response to a panel of 21 opioid peptides but still signaled normally in response to small molecule drugs such as spiradoline. Activation of a RASSL by spiradoline also caused proliferation of rat-1a tissue culture cells. These data provide evidence that G protein-coupled receptors can be made into RASSLs. The potential in vivo applications for RASSLs include the positive enrichment of transfected cells and the development of new animal models of disease. PMID- 9419380 TI - Obesity and hyperleptinemia in metallothionein (-I and -II) null mice. AB - Metallothionein (MT) has several putative roles in metal detoxification, in Zn and Cu homeostasis, in scavenging free radicals, and in the acute phase response. Mice of mixed 129/Ola and C57BL/6J background with targeted disruption of MT-I and MT-II genes are more sensitive to toxic metals and oxidative stress. We noted that these animals were larger than most strains of mice, and we systematically studied aspects of their physiology and biochemistry relating to energy metabolism. During the first 2 weeks after weaning, the growth rates of MT-null and C57BL/6J mice were similar, but the transgenic mice became significantly heavier at age 5-6 weeks. At age 14 weeks, the body weight and food intake of MT null mice was 16 and 30% higher, respectively, compared with C57BL/6J mice. Most 22- to 39-week-old male MT-null mice were obese, as shown by increased fat accretion, elevated obese (ob) gene expression, and high plasma leptin levels, similar to those recorded in Zucker fatty (fa/fa) rats. Seven-week-old MT-null mice also had significantly higher levels of plasma leptin and elevated expression of ob, lipoprotein lipase, and CCAAT enhancer binding protein alpha genes as compared with age-matched C57BL/6J mice. These observations indicate that abnormal accretion of body fat and adipocyte maturation is initiated at 5-7 weeks of age, possibly coincident with sexual maturation. Targeted disruption of MT-I and MT-II genes seems to induce moderate obesity, providing a new obese animal model. A link between MT and the regulation of energy balance is implied. PMID- 9419381 TI - The cyanobacterium Synechococcus resists UV-B by exchanging photosystem II reaction-center D1 proteins. AB - Current ambient UV-B levels can significantly depress productivity in aquatic habitats, largely because UV-B inhibits several steps of photosynthesis, including the photooxidation of water catalyzed by photosystem II. We show that upon UV-B exposure the cyanobacterium Synechococcus sp. PCC 7942 rapidly changes the expression of a family of three psbA genes encoding photosystem II D1 proteins. In wild-type cells the psbAI gene is expressed constitutively, but strong accumulations of psbAII and psbAIII transcripts are induced within 15 min of moderate UV-B exposure (0.4 W/m2). This transcriptional response causes an exchange of two distinct photosystem II D1 proteins. D1:1 is encoded by psbAI, but on UV-B exposure, it is largely replaced by the alternate D1:2 form, encoded by both psbAII and psbAIII. The total content of D1 and other photosystem II reaction center protein, D2, remained unchanged throughout the UV exposure, as did the content and composition of the phycobilisome. Wild-type cells suffered only slight transient inhibition of photosystem II function under UV-B exposure. In marked contrast, under the same UV-B treatment, a mutant strain expressing only psbAI suffered severe (40%) and sustained inhibition of photosystem II function. Another mutant strain with constitutive expression of psbAII and psbAIII was almost completely resistant to the UV-B treatment, showing no inhibition of photosystem II function and only a slight drop in electron transport. In Synechococcus the rapid exchange of alternate D1 forms, therefore, accounts for much of the cellular resistance to UV-B inhibition of photosystem II activity and photosynthetic electron transport. This molecular plasticity may be an important element in community-level responses to UV-B, where susceptibility to UV-B inhibition of photosynthesis changes diurnally. PMID- 9419382 TI - Rapid reorganization of resistance gene homologues in cereal genomes. AB - We used conserved domains in the major class (nucleotide binding site plus leucine-rich repeat) of dicot resistance (R) genes to isolate related gene fragments via PCR from the monocot species rice and barley. Peptide sequence comparison of dicot R genes and monocot R-like genes revealed shared motifs but provided no evidence for a monocot-specific signature. Mapping of these genes in rice and barley showed linkage to genetically characterized R genes and revealed the existence of mixed clusters, each harboring at least two highly dissimilar R like genes. Diversity was detected intraspecifically with wide variation in copy number between varieties of a particular species. Interspecific analyses of R like genes frequently revealed nonsyntenic map locations between the cereal species rice, barley, and foxtail millet although tight collinear gene order is a hallmark of monocot genomes. Our data suggest a dramatic rearrangement of R gene loci between related species and implies a different mechanism for nucleotide binding site plus leucine-rich repeat gene evolution compared with the rest of the monocot genome. PMID- 9419383 TI - A transcription activation system for regulated gene expression in transgenic plants. AB - A widely applicable promoter system is described that allows a gene of interest to be activated in specific plant tissues after a cross between defined transgenic lines. The promoter, pOp, consists of lac operators cloned upstream of a minimal promoter. No expression was detected from this promoter when placed upstream of a beta-glucuronidase (GUS) reporter gene in transgenic plants. Transcription from the promoter was activated by crossing reporter plants with activator lines that expressed a chimeric transcription factor, LhG4. This factor comprised transcription-activation domain-II from Gal4 of Saccharomyces cerevisiae fused to a mutant lac-repressor that binds its operator with increased affinity. When LhG4 was expressed from the CaMV 35S promoter, the spatial and quantitative expression characteristics of the 35S promoter were exhibited by the GUS reporter. The LhG4/pOp system may be used to study toxic or deleterious gene products, to coordinate the expression of multiple gene products, to restrict transgene phenotypes to the F1 generation, and to generate hybrid seed. The LhG4 system offers spatially regulated gene expression in the tissues of whole plants growing under normal conditions without the need for external intervention. It complements inducible expression systems that offer temporal control of gene expression in tissues that can be treated with inducing chemicals. PMID- 9419384 TI - Binding of an arm repeat protein to the kinase domain of the S-locus receptor kinase. AB - Screening of a yeast two-hybrid library for proteins that interact with the kinase domain of an S-locus receptor kinase (SRK) resulted in the isolation of a plant protein called ARC1 (Arm Repeat Containing). This interaction was mediated by the C-terminal region of ARC1 in which five arm repeat units were identified. Using the yeast two-hybrid system and in vitro binding assays, ARC1 was found to interact specifically with the kinase domains from SRK-910 and SRK-A14 but failed to interact with kinase domains from two different Arabidopsis receptor-like kinases. In addition, treatment with a protein phosphatase or the use of a kinase inactive mutant reduced or abolished the binding of ARC1 to the SRK-910 kinase domain, indicating that the interaction was phosphorylation dependent. Lastly, RNA blot analysis revealed that the expression of ARC1 is restricted to the stigma, the site of the self-incompatibility response. PMID- 9419385 TI - Cryptic speciation and recombination in the aflatoxin-producing fungus Aspergillus flavus. AB - Aspergillus flavus, like approximately one-third of ascomycete fungi, is thought to be cosmopolitan and clonal because it has uniform asexual morphology. A. flavus produces aflatoxin on nuts, grains, and cotton, and assumptions about its life history are being used to develop strategies for its biological control. We tested the assumptions of clonality and conspecificity in a sample of 31 Australian isolates by assaying restriction site polymorphisms from 11 protein encoding genes and DNA sequences from five of those genes. A. flavus isolates fell into two reproductively isolated clades (groups I and II). The lack of concordance among gene genealogies among isolates in one of the clades (group I) was consistent with a history of recombination. Our analysis included five strains of the closely related industrial fungus A. oryzae, all of which proved to be clonally related to group I. PMID- 9419386 TI - Deleterious mutation accumulation and the regeneration of genetic resources. AB - The accumulation of mildly deleterious mutations accompanying recurrent regeneration of plant germ plasm was modeled under regeneration conditions characterized by different amounts of selection and genetic drift. Under some regeneration conditions (sample sizes >/=75 individuals and bulk harvesting of seed) mutation accumulation was negligible, but under others (sample sizes <75 individuals or equalization of seed production by individual plants) mutation numbers per genome increased significantly during 25-50 cycles of regeneration. When mutations also are assumed to occur (at elevated rates) during seed storage, significant mutation accumulation and fitness decline occurred in 10 or fewer cycles of regeneration regardless of the regeneration conditions. Calculations also were performed to determine the numbers of deleterious mutations introduced and remaining in the genome of an existing variety after hybridization with a genetic resource and subsequent backcrossing. The results suggest that mutation accumulation has the potential to reduce the viability of materials held in germ plasm collections and to offset gains expected by the introduction of particular genes of interest from genetic resources. PMID- 9419387 TI - Letter recognition reveals pathways of second-order and third-order motion. AB - How are second-order (texture-defined) and third-order (pattern-tracking) motions processed in our brains? As shown here in the context of an ambiguous motion task involving a nominal second-order stimuli first devised by Werkhoven et al., [Werkhoven, P., Sperling, G. & Chubb, C. (1993) Vision Res. 33, 463-485.], the observers fell into two distinct groups based on the direction of perceived motion. The differences were interpreted in terms of the algorithms used to extract motion: one group by using a second-order motion process and the other by using a third-order motion process. This was investigated further using a dual task paradigm in which the interference between two tasks indicated the nature of processing involved. Observers who used third-order motion processing experienced interference with letter recognition and a more severe interference in dual third order motion tasks. Observers who used second-order motion processing experienced interference with another second-order motion detection but not with letter recognition. Insofar as task interference implies the need for attention, the complex interference effects and the apparently paradoxical interference effects of second-order motion perception imply that there are multiple forms of attention. Whether two tasks interfere depends on whether they require the same form of attention. Insofar as spatio-temporal processing is assumed to be carried out in the dorsal stream and pattern recognition in the ventral stream, the interference patterns suggest that second-order motion may be computed entirely in the dorsal stream, and third-order motion may involve two computational processes, one of which shares computational resources with the letter recognition task in the ventral stream. PMID- 9419388 TI - Photoreceptor guanylate cyclases: a review. AB - Almost three decades of research in the field of photoreceptor guanylate cyclases are discussed in this review. Primarily, it focuses on the members of membrane bound guanylate cyclases found in the outer segments of vertebrate rods. These cyclases represent a new guanylate cyclase subfamily, termed ROS-GC, which distinguishes itself from the peptide receptor guanylate cyclase family that it is not extracellularly regulated. It is regulated, instead, by the intracellularly-generated Ca2+ signals. A remarkable feature of this regulation is that ROS-GC is a transduction switch for both the low and high Ca2+ signals. The low Ca2+ signal transduction pathway is linked to phototransduction, but the physiological relevance of the high Ca2+ signal transduction pathway is not yet clear; it may be linked to neuronal synaptic activity. The review is divided into eight sections. In Section I, the field of guanylate cyclase is introduced and the scope of the review is briefly explained; Section II covers a brief history of the investigations and ideas surrounding the discovery of rod guanylate cyclase. The first five subsections of Section III review the experimental efforts to quantify the guanylate cyclase activity of rods, including in vitro and in situ biochemistry, and also the work done since 1988 in which guanylate cyclase activity has been determined. In the remaining three subsections an analytical evaluation of the Ca2+ modulation of the rod guanylate cyclase activity related to phototransduction is presented. Section IV deals with the issues of a biochemical nature: isolation and purification, subcellular localization and functional properties of rod guanylate cyclase. Section V summarizes work on the cloning of the guanylate cyclases, analysis of their primary structures, and determination of their location with in situ hybridization. Section VI summarizes studies on the regulation of guanylate cyclases, with a focus on guanylate cyclases activating proteins. In Section VII, the evidence about the localization and functional role of guanylate cyclases in other retinal cells, especially in "on-bipolar" cells, in which guanylate cyclase most likely plays a critical role in electrical signaling, is discussed. The review concludes with Section VIII, with remarks about the future directions of research on retinal guanylate cyclases. PMID- 9419389 TI - D-glucose but not insulin reduces N-formyl-methionyl-leucyl-phenylalanine (fMet Leu-Phe)-induced shape changes in suspended human neutrophils. AB - The effects of glucose (5-25 mM) and insulin concentration (40-320 microU/ml) on the cell shape of neutrophil granulocytes from healthy humans were studied. Both non-activated and N-formyl-methionyl-leucyl-phenylalanine (fMet-Leu-Phe) activated neutrophils in suspension were used as a model for initial chemotactic activation of neutrophil locomotion. D-glucose, but not the non-metabolizable analogue 3-O-methyl-D-glucose, dose-dependently reduced the fMet-Leu-Phe-induced (10(-8)M) neutrophil elongation. Insulin, either alone or in combination with 25 mM D-glucose, was without effect on the fMet-Leu-Phe-induced neutrophil elongation. Furthermore, the inhibitory effect of D-glucose was observed already after 1 min of exposure to D-glucose and fMet-Leu-Phe. D-glucose diminished the fraction of neutrophils with elongated locomotor shape by changing it into an irregular cell shape, suggesting that at least part of the D-glucose effect could be associated with mechanisms determining the typical locomotor shape. The present results suggest that D-glucose through its metabolism, but without the involvement of insulin, reduces chemotactically induced elongation to a locomotor neutrophil shape, and thus neutrophil motility, and that this effect of glucose appears prior to adhesion. This glucose-induced inhibition of the neutrophil chemotactic response may be involved in the neutrophil deficiency seen in diabetes mellitus. PMID- 9419390 TI - Effects of D-glucose on chemokinesis and resting production of reactive oxygen species in neutrophil granulocytes of lean or obese-hyperglycemic mouse. AB - The response to D-glucose (0-21 mM) was studied in neutrophil granulocytes from obese, hyperglycemic and hyperinsulinemic Umea ob/ob mice and their lean, littermate controls in order to further elucidate the effects of in vivo and in vitro hyperglycemia on neutrophil function. Neutrophil random locomotion on glass and neutrophil resting luminol-enhanced chemiluminescence in cell suspension were studied. Random locomotion was stimulated by D-glucose in neutrophils from both Umea ob/ob and control mice but the locomotive activity in Umea ob/ob mouse neutrophils was significantly higher than that found in the controls at 4-21 mM glucose. In both types of mice, the stimulatory effect of D-glucose on random locomotion was diminished at 21 mM glucose (not significantly different from that at 0 mM glucose). Resting chemiluminescence from mouse neutrophils was also stimulated by glucose but here the magnitude of response was similar in neutrophils from both types of mice. These results indicate that chronic hyperglycemia and hyperinsulinemia in the Umea ob/ob mouse may be associated with an increased neutrophil random locomotive activity but a similar resting production of reactive oxygen species, as compared with neutrophils from control mice at physiological and hyperglycemic glucose concentrations in vitro. PMID- 9419391 TI - Erythrocyte membrane digoxin-sensitive (Na(+)-K+)-ATPase of non-insulin dependent diabetic humans. AB - Erythrocyte plasma membranes of non-insulin dependent diabetic humans (NIDDM) and healthy humans were prepared by hypotonic lysis. The specific activity of (Na(+) K+)-ATPase of NIDDM membranes, both in the absence and presence of digoxin were lower than the specific activity of normal enzymes (83.6 percent and 74.0 percent of the normal enzyme respectively). Addition of digoxin decreased the activity of this enzyme (38.0 percent in NIDDM and 30.0 percent in normal enzyme). Although the affinity of the pump for ATP was similar in both membranes of NIDDM and normal humans (K(m) for ATP = 19.9 +/- 0.24 microM ATP and 20.0 +/- 0.21 microM ATP respectively), the Vmax of NIDDM membranes was more than 20 percent lower than that of the normal enzyme. The specific activity of Mg(2+)-dependent Ca(2+) pumping ATPase (Ca(2+)-Mg(2+)-ATPase) of NIDDM membrane was lower than 80 percent of the specific activity of the normal enzymes. While the affinity of the pump for ATP was lower in the membranes of NIDDM (Km for ATP = 50.0 +/- 4.3 microM ATP) in comparison to normal membranes (Km for ATP = 63.1 +/- 38 microM ATP), the Vmax of NIDDM membranes was similar to the normal enzyme. Altogether, these findings suggest that both the (Na(+)-K+)-ATPase and Ca(2+)-pumping ATPase of NIDDM membranes are less functional than the enzymes in normal erythrocytes. PMID- 9419392 TI - Etiology, natural history, management and molecular genetics of hereditary nonpolyposis colorectal cancer (Lynch syndromes): genetic counseling implications. AB - We estimate that 5-10% of virtually all forms of cancer are due to a primary hereditary etiology. However, a hereditary cancer diagnosis is often missed because the family history of cancer is given short shrift in medical practice. Hereditary nonpolyposis colorectal cancer (HNPCC) certainly fits this estimate, although some studies suggest that a minimum of 2% with a range as high as 10% of the total colorectal cancer burden is due to HNPCC. Mutations in one of the four mismatch repair genes, i.e., hMSH2, hMLH1, hPMS1, and hPMS2, account for about 70% of HNPCC kindreds. Other germ-line mutations are likely to be identified to account for the remainder of HNPCC patients. By far the most common HNPCC mutations involve hMSH2 and hMLH1, with hPMS1 and hPMS2 accounting for only about 3% of such families. Prior to these molecular genetic discoveries, the genetic counselor could only provide the patient with an estimate of a 50% likelihood of manifesting HNPCC based on the counselee having one or more first-degree relatives manifesting syndrome cancers in their direct genetic lineage. Because DNA testing has become available in families with known mutations, we have provided pretest group education in the form of a family information service with intensive education about the natural history, genetic risk, surveillance, and options for management of HNPCC, as well as discussion of the potential for fear, anxiety, apprehension, and insurance or employer discrimination that might impact on this DNA testing. Following informed consent, these relatives were then counseled on a one-to-one basis. Using DNA-based genetic counseling involving hMSH2 or hMLH1, we have provided this service to four extended HNPCC kindreds. Details of this genetic counseling experience on these four kindreds will be discussed. PMID- 9419393 TI - Effect of paternal and maternal cancer on cancer in the offspring: a population based study. AB - The Family-Cancer Database was constructed from the nationwide Swedish registries to include more than 30,000 cancers in offspring diagnosed at ages 15-51 years and their parents. Cancer risk in the offspring was increased about 1.10 times when the father had cancer, whereas no increase was noted when the mother had cancer. If both parents had cancer, the risk for sons was 1.39 and for daughters, 1.34. Familial aggregation between parents and offspring was observed for 5 concordant and 14 discordant cancer sites and 10 parental sites at which all cancer was increased in the offspring. The concordant sites between the parent and offspring were colorectum, breast, melanoma, skin (squamous cell carcinoma), and thyroid. The aggregation at discordant sites in the parents and the offspring included stomach-breast, colorectum-salivary glands, colorectum-breast, colorectum-lymphoma, colorectum-leukemia, liver-breast, pancreas-breast, breast melanoma, ovary-breast, prostate-breast, prostate-cervix, prostate-multiple myeloma, kidney-melanoma, and nervous tissue-melanoma. In most of these combinations, cancer in the second parent increased the risk to the offspring. The present results on young and middle-aged adults suggest that cancer in both parents increases cancer risk in the offspring at many sites. Chance and environmental effects may explain some of the results, whereas true genetic factors probably contribute to most of the findings. The molecular genetic explanation may be that rare dominant single genes increase susceptibility at many sites or that overlapping sets of genes control susceptibility at multiple sites. PMID- 9419394 TI - Age-period-cohort modeling of colorectal cancer incidence and mortality in Spain. AB - Spain registers a much lower rate of colorectal cancer incidence and mortality than do other European countries, yet the rises observed in the adjusted rates over recent decades led us to attempt to monitor the trends over time using Poisson log-linear models. Incidence data were furnished by the Zaragoza and Navarre population-based cancer registries, whereas mortality data corresponded to Spain as a whole. For trend evaluation purposes, we made use of invariant parameters from age-period-cohort models (net drift and curvature) and a restriction of the cohort-effect slope range. The results suggest the presence of a marked rise in incidence of colorectal cancer for both sexes and across all age groups in the provinces studied. The rise in mortality was less pronounced than the rise in incidence and seemed to coincide with a marked cohort effect present throughout the study period. Both in incidence and mortality, the increases were more pronounced among men. When studied jointly, incidence and mortality trends tend to be complementary, rendering an approach of this nature especially important in sites with better survival, such as the case in point. PMID- 9419395 TI - Diabetes mellitus and colorectal cancer risk. AB - The relationship between diabetes mellitus and the risk of colorectal cancer was investigated in a multicenter case-control study, conducted in Italy between 1992 and 1996 on 1225 cases of incident, histologically confirmed colon cancer, 728 cases of rectal cancer, and 4154 controls, who were in the hospital for acute, nonneoplastic diseases. Overall, 66 (5.4%) cases of colon cancer, 50 (6.9%) cases of rectal cancer, and 185 (4.4%) controls reported a history of diabetes. The corresponding multivariate odds ratios (ORs) were 1.2 [95% confidence interval (CI), 0.8-1.6] for colon, 1.5 (95% CI, 1.1-2.2) for rectal, and 1.3 (95% CI, 1.0 1.6) for all colorectal cancers. No association was observed for subjects who were diagnosed with diabetes at ages of < 40 years (7 cases and 27 controls, OR = 0.9). The OR was 1.4 (95% CI, 1.1-1.7) for subjects who were diagnosed with diabetes at ages of > or = 40 years and were likely to have non-insulin-dependent diabetes. The association was also stronger (OR = 1.6; 95% CI, 1.1-2.3) among subjects whose diabetes was diagnosed 10 or more years in advance and who were > or = 60 years old at the time of colorectal cancer diagnosis. None of the other covariates, including sex, education, body mass index, physical activity, energy intake, alcohol drinking, and fiber intake, showed any appreciable modifying effect. Thus, this uniquely large case-control study of colorectal cancer confirms that subjects with non-insulin-dependent diabetes mellitus have a slightly increased risk of colorectal cancer. More importantly, allowance for a large number of identified potential confounding factors, including body mass index, diet, and physical activity, could not explain the excess colorectal cancer risk among subjects with diabetes mellitus. These findings have plausible biological correlations because insulin-like-growth factor-I is a promoter of colon tumor cell growth in vitro. PMID- 9419396 TI - Colorectal epithelial cell proliferative kinetics and risk factors for colon cancer in sporadic adenoma patients. AB - Colorectal epithelial cell proliferative kinetics are altered in patients at increased risk for colon cancer: proliferation rates [labeling index (LI)] are higher and there is a shift of the proliferative zone from one confined to the lower 60% of the colonic crypt to one that includes the entire crypt (higher phi(h)). To assess factors associated with LI and phi(h), we performed a cross sectional analysis using baseline rectal mucosal biopsies from sporadic adenoma patients participating in a chemoprevention trial. Biopsies (taken without preparatory cleansing) were taken 10 cm above the level of the anus, and proliferation was assessed by detection of endogenous S-phase-associated proliferating cell nuclear antigen by immunohistochemical methods. High-quality, scorable biopsies were obtained for 115 patients, and using analysis of covariance and multiple linear regression, the LI and phi(h) were evaluated in relation to diet and other lifestyle factors, demographics, anthropometrics, family history of colon cancer, and polyp history. Statistically significant findings included the following: (a) The LI for those in the upper versus the lowest tertile of vegetable and fruit consumption was, proportionately, 35% lower (3.4% versus 5.3%; P < 0.001); for vitamin supplement users versus nonusers, it was 36% lower (3.3 versus 5.2%; P < 0.001); for recurrent versus incident polyp patients, it was 36% higher (6.2 versus 4.0%; P < 0.001); and for those with rectal polyps only versus those with colon polyps only, it was 28% higher (6.0 versus 4.3%; P = 0.05); and (b) the phi(h) for those in the upper versus the lowest tertile of sucrose consumption was, proportionately, 48% higher (7.1% versus 3.7%; P = 0.01). These results indicate that (a) colorectal epithelial cell proliferation rates are higher in recurrent adenoma patients than in incident adenoma patients and in patients with rectal adenomas only versus those with colon adenomas only, but they are lower in patients with higher intakes of vegetables and fruit and in those who take vitamin/mineral supplements, and (b) the distribution of proliferating cells is shifted toward more inclusion of the upper 40% of the crypt in patients with higher intakes of sucrose. The pattern of positive, negative, and null associations of potential risk factors with cell proliferation is similar to that commonly found with colonic neoplasms. PMID- 9419397 TI - Calcium and colorectal epithelial cell proliferation in ulcerative colitis. AB - In persons at higher risk for colon cancer (e.g., those with sporadic adenoma or ulcerative colitis), compared to those at lower risk, colonic epithelial cell proliferation kinetics are altered. We have shown previously that calcium supplementation appears to normalize the distribution of proliferating cells without affecting the proliferation rate in the colorectal mucosa of sporadic adenoma patients. In a pilot randomized, double-blind, placebo-controlled, clinical trial conducted concurrently with our previously published sporadic adenoma trial, we tested whether calcium supplementation can also modulate cell proliferation kinetics in patients with ulcerative colitis. Ulcerative colitis patients (n = 31) were randomized to placebo or 2.0 g of supplemental calcium daily. Colorectal epithelial cell proliferation was determined by immunohistochemical detection of proliferating cell nuclear antigen labeling of cells in "nonprep" rectal biopsies taken at randomization and after 2 months treatment. All biopsies were scored by one reviewer. Differences in mean follow up minus baseline labeling index (LI; the proportion of colon crypt epithelial cells that were labeled) and in the phi(h) (proportion of labeled cells that were in the upper 40% of the crypts) were compared with analysis of covariance. Pill taking adherence was 97%. Biopsy-scoring reliability was high (r = 0.89). The pooled baseline LI and phi(h) were 6.3% and 5.6%, respectively. The LI in the calcium group decreased by 0.5% (proportionately, 3%) more than in the placebo group (P = 0.91). Similarly, the phi(h) in the calcium group decreased by 0.3% (proportionately, 10%) more than in the placebo group (P = 0.85). This pilot study does not suggest that 2.0 g of calcium as calcium carbonate daily can substantially normalize either the rate or distribution of proliferating cells over a 2-month period in the colon crypts of patients with ulcerative colitis; a more definitive answer to the question of whether calcium may be effective would require a study with a larger sample size and/or other study design modifications. PMID- 9419398 TI - Dietary iron and recurrence of colorectal adenomas. AB - Previous research suggests that iron acts as a prooxidant to increase the risk of colorectal neoplasia. This study examined effects of dietary intake of iron on colorectal adenoma recurrence using data from an antioxidant clinical trial. All subjects were free of polyps at study entry but had at least one adenoma removed within the 3 months before enrollment. Follow-up colonoscopies were conducted after 1 and 4 years. Patients who developed one or more adenomatous polyps between years 1 and 4 were classified as cases; all others were controls. Dietary iron intake at baseline and at the end of the study was estimated from self administered food frequency questionnaires and averaged together for each subject, energy-adjusted, and categorized into quartiles. Odds ratios were adjusted for age, center, sex, calories, treatment group, and alcohol, fiber, folate, and fat intakes in unconditional logistic regression analysis. Dietary iron was inversely associated with adenoma risk, although risk did not decrease monotonically with increasing intake. Odds ratios comparing second, third, and fourth quartiles to the lowest quartile were 0.61 [95% confidence interval (CI), 0.37-1.02], 0.80 (95% CI, 0.45-1.44), and 0.37 (95% CI, 0.19-0.73), respectively. A limited examination showed no clear evidence that use of iron supplements affected risk of recurrence in this study population. This study provides evidence against the hypothesis that recent dietary intake of iron increases risk for colorectal adenomas. However, these results may reflect the presence of other dietary factors found in combination with iron. PMID- 9419399 TI - The aminothiol WR-1065 protects T lymphocytes from ionizing radiation-induced deletions of the HPRT gene. AB - Aminothiols, such as WR-2721 and its active free thiol, WR-1065, reduce mutations from ionizing radiation in exponentially growing cells. In this study, human noncycling G0 T lymphocytes were exposed in vitro to gamma-irradiation in the presence or absence of WR-1065. The five treatment groups were: (a) control; (b) treatment with 4 mM WR-1065; (c) treatment with 3 Gy of gamma-radiation, from a 137Cs source; and (d) and (e) treatment with WR-1065 30 min prior to or 3 h after 3 Gy of gamma-irradiaiton, respectively. A total of 224 cloned HPRT mutants representing 179 independent mutations were analyzed for genetic alterations using multiplex PCR. Ionizing radiation alone significantly increased the percentage of mutations with gross structural alterations compared to controls (P = 0.02). Although the frequency of such large structural mutations was not different from control cells treated with WR-1065 alone, this aminothiol significantly reduced their frequency among irradiated mutants (P = 0.01) when the radioprotector was present during the irradiation. Addition of WR-1065 3 h postirradiation also greatly reduced the percentage of gross structural alterations; however, due to small numbers, this was not statistically significant. This is the first demonstration that the antimutagenicity of WR-1065 in human cells specifically protects against these kinds of large-scale DNA alterations induced by ionizing radiation. WR-1065 and similar aminothiol compounds may afford protection against radiation-induced mutations through polyamine-like processes, e.g., stabilization of chromatin structure, inhibition of cell proliferation, and influences on DNA repair systems. PMID- 9419400 TI - Acidic urine pH is associated with elevated levels of free urinary benzidine and N-acetylbenzidine and urothelial cell DNA adducts in exposed workers. AB - We evaluated the influence of urine pH on the proportion of urinary benzidine (BZ) and N-acetylbenzidine present in the free, unconjugated state and on exfoliated urothelial cell DNA adduct levels in 32 workers exposed to BZ in India. Postworkshift urine pH was inversely correlated with the proportions of BZ (r = -0.78; P < 0.0001) and N-acetylbenzidine (r = -0.67; P < 0.0001) present as free compounds. Furthermore, the average of each subject's pre- and postworkshift urine pH was negatively associated with the predominant urothelial DNA adduct (P = 0.0037, adjusted for urinary BZ and metabolites), which has been shown to cochromatograph with a N-(3'-phosphodeoxyguanosin-8-yl)-N'-acetylbenzidine adduct standard. Controlling for internal dose, individuals with urine pH < 6 had 10 fold higher DNA adduct levels compared to subjects with urine pH > or = 7. As reported previously, polymorphisms in NAT1, NAT2, and GSTM1 had no impact on DNA adduct levels. This is the first study to demonstrate that urine pH has a strong influence on the presence of free urinary aromatic amine compounds and on urothelial cell DNA adduct levels in exposed humans. Because there is evidence that acidic urine has a similar influence on aromatic amines derived from cigarette smoke, urine pH, which is influenced by diet, may be an important susceptibility factor for bladder cancer caused by tobacco in the general population. PMID- 9419401 TI - Evaluation of a food frequency questionnaire-food composition approach for estimating dietary intake of inorganic arsenic and methylmercury. AB - Inorganic arsenic intake in 969 men and women and methylmercury intake in 785 men and women from across the United States were assessed by a semiquantitative food frequency questionnaire, in combination with a database for the content of those elements in foods, and by toenail concentrations of arsenic and mercury. In addition, empirical weights for foods on the dietary questionnaire were derived from multivariate regression models to estimate associations between diet and toenail arsenic and mercury levels, independent of the assumptions about inorganic arsenic and methylmercury in foods, which are based upon limited residue measurements. The use of empirical weights significantly improved the correlation of arsenic consumption with toenail arsenic levels (r = 0.33, P = 0.0001), compared with the weak correlation obtained using the food residue method to calculate intake (r = 0.15, P = 0.0001). Mercury consumption computed using empirical weights yielded a significant correlation with toenail arsenic (r = 0.42, P = 0.001), similar to the correlation using energy-adjusted intake calculated from food residue tables (r = 0.35, P = 0.001). These results illustrate the potential use of empirically derived weights for foods in estimating toenail levels of selected heavy metals and support the validity of published food residue data that are used to estimate mercury consumption. PMID- 9419402 TI - Decrease in bladder cell micronucleus prevalence after intervention to lower the concentration of arsenic in drinking water. AB - Epidemiological studies performed in Taiwan, Argentina, and Chile suggest that ingestion of arsenic (As) may cause bladder cancer. Because of these findings, we previously investigated the relationship between As ingestion and genetic damage to the urothelium in two cross-sectional biomarker studies, one in Nevada and one in Chile. In both studies, we found that increased levels of micronucleated cells (MNCs) in exfoliated bladder cells were associated with elevated concentrations of As in drinking water, suggesting that As induces genetic damage to bladder cells. To further investigate this relationship, we conducted an intervention study in a subset of highly exposed men (n = 34) from the cross-sectional study in Chile. Subjects whose usual source of water contained about 600 micrograms/liter As were supplied with water lower in As (45 micrograms/liter) for 8 weeks, allowing ample opportunity for renewal and exfoliation of bladder epithelial cells. Mean urinary As levels decreased during the intervention from 742 to 225 micrograms/liter. Bladder MNC prevalence also decreased from 2.63 MNCs/1000 cells preintervention to 1.79 MNCs/1000 cells postintervention (P < 0.05). When the analysis was limited to individuals previously having subcytotoxic urinary As levels (< 700 micrograms/liter), the change between pre- and postintervention MNC was more pronounced: the level decreased from 3.54 to 1.47 MNCs/1000 cells, respectively (P = 0.002). Among smokers, MNC prevalences decreased from 4.45 MNCs/1000 cells preintervention to 1.44 MNCs/1000 cells postintervention (P = 0.002). Among nonsmokers, the decrease was much smaller: 2.04 MNCs/1000 cells preintervention to 1.90 MNCs/1000 cells postintervention (P = 0.25), suggesting that smoker's bladder cells could be more susceptible to genotoxic damage caused by As. The reduction in bladder MNC prevalence with reduction in As intake provides further evidence that As is genotoxic to bladder cells. PMID- 9419403 TI - Identification of concurrent germ-line mutations in hMSH2 and/or hMLH1 in Japanese hereditary nonpolyposis colorectal cancer kindreds. AB - We analyzed microsatellite instability, alterations of the polyadenine tract in TGF-beta RII (transforming growth factor beta type II receptor gene), and mutations of hMSH2 and hMLH1 in 32 patients with familial colorectal cancer (29 kindreds) fulfilling the clinical criteria for hereditary nonpolyposis colorectal cancer (HNPCC), defined at the 34th Annual Meeting of Japanese Society for Cancer of the Colon and Rectum (Tokushima, Japan, 1991), including five kindreds fulfilling the Amsterdam criteria. Eighteen of 32 (56%) cases were replication error positive (RER+) at two or more microsatellite loci analyzed. The clinicopathological characteristics of RER+ cases corresponded well with those reported previously. Eleven of 18 RER+ cases showed RER+ at most of the microsatellite loci examined. Among these 11 cases (10 kindreds), 3 kindreds fulfilled the Amsterdam criteria and 7 kindreds did not. For these 10 kindreds, germ-line mutations in hMSH2 and hMLH1 were detected for 6 kindreds by PCR-SSCP analysis and direct sequencing. Only two of these six fulfilled the Amsterdam criteria; more than one germ-line mutation was detected in hMSH2 and/or hMLH1. Specifically, two point mutations of hMSH2 were detected in two kindreds, one point mutation of both hMSH2 and hMLH1 was detected in one kindred, two point mutations of hMSH2 and one point mutation of hMLH1 were detected in one kindred, and two point mutations of hMLH1 and one point mutation of hMSH2 were detected in one kindred. In addition, 19 of 26 (74%) cancer lesions of these 11 cases with the RER phenotype showed alterations of the polyadenine tract in TGF-beta RII. From our data, although seven kindreds did not fulfill the Amsterdam criteria, we considered them as HNPCC. Therefore, we suggest that the "Japanese criteria" have the advantage of being able to detect more HNPCC kindreds from borderline HNPCC kindreds. PMID- 9419404 TI - Diet, Helicobacter pylori, and p53 mutations in gastric cancer: a molecular epidemiology study in Italy. AB - A series of 105 gastric cancer (GC) cases with paraffin-embedded specimens interviewed in a previous population-based case-control study conducted in a high risk area around Florence, Italy, was examined for the presence of p53 mutations. Overall, 33 of 105 cases had a mutation (p53+) identified by single-strand conformational polymorphism and confirmed by sequencing (Y-H. Shiao et al., submitted for publication). p53+ cases had a more traditional dietary pattern (i.e., corn meal mush, meat soup, and other homemade dishes) and reported less frequent consumption of raw vegetables (particularly lettuce and raw carrots). A positive association with a high nitrite intake and a negative association with raw vegetables and diffuse type histology persisted in a multivariate analysis. In addition, p53+ cases tended to be located in the upper portion of the stomach and to be associated with advanced age and blood group A. No relation was found between the presence of p53 mutations and histologically defined Helicobacter pylori infection, smoking history, family history of gastric cancer, education, and social class. Of the 33 p53+ cases, 19 had G:C-->A:T transitions at CpG sites. These tumors tended to occur in females and in association with H. pylori infection but not other risk factors. The remaining 14 cases with a p53 mutation had mainly transversions but also two deletions and two transitions at non-CpG sites. These tumors showed a strong positive association with a traditional dietary pattern and with the estimated intake of selected nutrients (nitrite, protein, and fat, particularly from animal sources). The findings of this case case analysis suggest that p53 mutations at non-CpG sites are related to exposure to alkylating compounds from diet, whereas p53 mutations at CpG sites might be related to H. pylori infection. PMID- 9419405 TI - MUC6 gene polymorphism in healthy individuals and in gastric cancer patients from northern Portugal. AB - Mucins exhibit a high degree of genetic polymorphism because of the presence of a variable number of tandem repeats. The aims of this work were to describe the MUC6 gene polymorphism in the Portuguese population and to evaluate whether MUC6 gene polymorphism was involved in individual susceptibility to gastric cancer development, as observed previously for the MUC1 gene. We found that the 10 alleles identified in the population of blood donors (n = 376), by Southern blot analysis, were also found in gastric cancer patients (n = 157). However, significant differences in allelic frequencies between the two populations were observed for 4 of the 10 alleles, in agreement with those described previously for the MUC1 gene; the largest allele was more frequent in blood donors, and smaller alleles were more frequent in gastric cancer patients. Our results suggest that MUC6 gene polymorphism is involved in the predisposition to gastric carcinoma development. PMID- 9419406 TI - Susceptibility to lung cancer in light smokers associated with CYP1A1 polymorphisms in Mexican- and African-Americans. AB - The gene-environment associations between potential carcinogenic agents modified by polymorphisms in the cytochrome P450 1A1 (CYP1A1) gene and lung cancer risk were assessed in a hospital-based case-control study composed of African- and Mexican-Americans. The study involved 171 cases and 295 controls identified from the greater Houston and San Antonio metropolitan areas. Both the exon 7 and MspI polymorphisms were analyzed by RFLP of PCR-amplified DNA, and in addition, the African-American-specific polymorphism was assayed for subjects who reported that they were African-American. Logistic regression analysis was performed to assess the association between each of the CYP1A1 polymorphisms and lung cancer, adjusting for the matching variables (age, sex, ethnicity) and other potential risk factors. Interactions between pack-years smoked, CYP1A1 genotypes, and case status were also evaluated. The variant allele frequencies did not differ by case status, but the distributions of genotypes were strikingly different by ethnicity. In addition, both the exon 7 and MspI polymorphisms, but not the African-American-specific polymorphism, were modified by the amount of cigarette consumption measured in pack-years. An approximate 2-fold increase in lung cancer risk among individuals with one or more of the variant alleles was observed among light smokers (defined as having smoked < or = 30 pack-years). The respective risk ratios for the exon 7 and MspI polymorphisms were 2.26 (95% confidence interval, 0.82-6.26) and 2.03 (95% confidence interval, 1.03-4.01) at low smoking dose. No such increase in risk was found among heavy smokers (> 30 pack-years). This phenomenon at low smoking dose was also observed when the two common polymorphisms were combined, which resulted in was a progressive increase in risk with an increasing number of variant alleles. These results indicate that at low smoking levels, the MspI and exon 7 CYP1A1 genetic polymorphisms confer susceptibility to lung cancer. PMID- 9419407 TI - Smoking, alcohol, coffee, and tea intake and incidence of cancer of the exocrine pancreas: the Iowa Women's Health Study. AB - To assess the relationship of smoking and coffee, tea, and alcohol intake to the risk of cancer of the exocrine pancreas, analyses were performed using data from a prospective cohort study of 33,976 postmenopausal Iowa women who responded to a mailed questionnaire in 1986 and were followed through 1994 for cancer incidence and total mortality. At baseline, information on cigarette smoking, consumption of tea, coffee, and alcoholic beverages, and other dietary and lifestyle factors was obtained. Age-adjusted relative risks of pancreatic cancer (n = 66 cases) showed a dose-response association with smoking. Those with fewer than 20 pack years and those with 20 or more pack-years of smoking exposure were 1.14 (95% confidence interval, 0.53-2.45) and 1.92 (95% confidence interval, 1.12-2.30) times more likely, respectively, to develop pancreatic cancer than were nonsmokers. Current smokers were twice as likely as were nonsmokers to develop pancreatic cancer. Relative risks of pancreatic cancer increased with the amount of alcohol consumed (Ptrend = 0.11) after adjustment for age, smoking status, and pack-years of smoking. Relative risks of pancreatic cancer according to alcoholic beverage intake were as strong among never-smokers as they were in the total cohort. After the data were adjusted for age, smoking status, and pack-years of smoking, there was a statistically significant 2-fold (95% confidence interval, 1.08-4.30) elevated risk of pancreatic cancer for those who drank > 17.5 cups of coffee per week, compared to those who consumed < 7 cups/week; among never smokers, the relative risks across coffee intake categories were still positive but were attenuated somewhat (P trend = 0.17). Tea intake was not related to cancer incidence. In summary, these findings provide evidence of an association of both alcoholic beverage and coffee consumption with pancreatic cancer incidence that is independent of age and cigarette smoking. PMID- 9419408 TI - Aggressiveness of colon carcinoma in blacks and whites. National Cancer Institute Black/White Cancer Survival Study Group. AB - Black patients with colon cancer in the Black/White Cancer Survival Study were found to have a poorer survival than white patients. More advanced-stage disease at diagnosis was the primary determinant, accounting for 60% of the excess mortality. After adjusting for stage, factors such as poverty, other socioeconomic conditions, and treatment did not further explain the remaining survival deficit. This study examined the aggressiveness of colon tumors in blacks and whites to explore its role in the racial survival differences. Tumor characteristics of 703 cases of newly diagnosed invasive colon adenocarcinoma were centrally evaluated by a gastrointestinal pathologist, blinded in regard to the age, race, and sex of the patients. Blacks were less likely to have poorly differentiated (grade 3) tumors [odds ratio (OR), 0.44; 95% confidence interval, 0.22-0.88] and lymphoid reaction (OR, 0.49; 95% confidence interval, 0.26-0.90) when compared with whites. These black/white (B/W) differences remained statistically significant after adjusting for age, sex, metropolitan area, summary stage, socioeconomic status, body mass index, and health care access and utilization. In addition, blacks were less likely to have high-grade (grade 3) nuclear atypia, mitotic activity, and tubule formation, although these ORs did not reach a statistical significance level of 0.05. Similar B/W differences were observed for patients with advanced disease but not with early stage. Comparison by anatomical subsite showed that blacks had statistically significantly better differentiated tumors for cancers of the proximal and transverse colon but not for the distal. No racial differences were found for blood vessel and lymphatic invasion, necrosis, fibrosis, and mucinous type of histology. The findings, therefore, are the opposite of those hypothesized. After adjusting for stage, more aggressive tumor characteristics do not explain the adverse survival differential in blacks. This suggests that there may be racial differences in environmental exposure, and that the intensity and mode of delivery of carcinogen insult as well as host susceptibility may differ by race and anatomical subsite. Future studies should explore the B/W differences in tumor biology using molecular markers that precede the conventional histological parameters evaluated here. PMID- 9419409 TI - Detection of K-ras mutations in resected primary leiomyosarcoma. AB - Mutation of the K-ras oncogene occurs frequently in human malignancy. However, there are few reports concerning K-ras mutations in soft-tissue sarcoma, including leiomyosarcoma. We therefore designed a study to determine the prevalence of mutations in the first exon of K-ras in leiomyosarcoma and to evaluate its prognostic potential. Fifty-one leiomyosarcomas were reviewed, and their diagnoses were confirmed on pathological review. Tissue blocks were retrieved, and new sections were prepared for confirmation of diagnosis. Additional tissue sections were used for DNA isolation. PCR and denaturing gradient gel electrophoresis (DGGE) were used to detect K-ras mutations in the first exon of genomic DNA isolated from the specimens. Seven (14%) K-ras mutations were detected using DGGE. Subsequent sequencing of the K-ras gene from each of the mutated tumors confirmed the DGGE results in each case. The median survival for patients whose tumors did not contain mutations of K-ras was 42 months (n = 42) versus 25 months (n = 7) for those with mutations (P = 0.06). However, patients with stages I and II tumors had a median survival of 82 months (n = 28) compared to 28 months for those with stages III and IV disease (n = 20, P = 0.02). The results suggest that K-ras codon 12 mutations are uncommon in leiomyosarcoma; however, when such mutations are found, there is a trend toward worse survival. Furthermore, the data confirm that stage is a significant prognostic indicator. PMID- 9419410 TI - Squalene, olive oil, and cancer risk: a review and hypothesis. AB - Epidemiological studies of breast and pancreatic cancer in several Mediterranean populations have demonstrated that increased dietary intake of olive oil is associated with a small decreased risk or no increased risk of cancer, despite a higher proportion of overall lipid intake. Experimental animal model studies of high dietary fat and cancer also indicate that olive oil has either no effect or a protective effect on the prevention of a variety of chemically induced tumors. As a working hypothesis, it is proposed that the high squalene content of olive oil, as compared to other human foods, is a major factor in the cancer risk reducing effect of olive oil. Experiments in vitro and in animal models suggest a tumor-inhibiting role for squalene. A mechanism is proposed for the tumor inhibitory activity of squalene based on its known strong inhibitory activity of beta-hydroxy-beta-methylglutaryl-CoA reductase catalytic activity in vivo, thus reducing farnesyl pyrophosphate availability for prenylation of the ras oncogene, which relocates this oncogene to cell membranes and is required for the signal transducing function of ras. PMID- 9419411 TI - Reliability of a flushing questionnaire and the ethanol patch test in screening for inactive aldehyde dehydrogenase-2 and alcohol-related cancer risk. AB - Molecular epidemiology of esophageal and upper aerodigestive tract cancers revealed that alcohol is more carcinogenic in persons with inactive aldehyde dehydrogenase-2 (ALDH2) than in those with active ALDH2. A simple questionnaire has been developed to screen for the facial flushing that occurs in persons with inactive ALDH2 when they drink even a single glass of beer. In this study, 266 of 284 consecutive male Japanese clinic patients (age > or = 50 years) completed the flushing questionnaire, and 239 underwent the ethanol patch test (a cutaneous model for the flushing response). Blinded genotyping showed inactive ALDH2 for 94.4% (102 of 108) of subjects who reported always flushing (early in their drinking history or currently) and for 47.7% (21 of 44) of those who reported sometimes flushing, whereas 95.6% (109 of 114) of subjects reporting that they never exhibited facial flushing had active ALDH2. When all three categories of flushing (current always, former always, and sometimes) were collapsed into one, the questionnaire's sensitivity and specificity for identifying inactive ALDH2 were 96.1 and 79.0%, respectively, compared with 72.4 and 71.4% for the ethanol patch test. The results suggest the utility of this simple flushing questionnaire in daily practice, as well as large-scale studies to assess cancer risks associated with drinking and ALDH2 and for activities aimed at preventing alcohol related cancer. PMID- 9419413 TI - Embryonic and adult expression patterns of the Tec tyrosine kinase gene suggest a role in megakaryocytopoiesis, blood vessel development, and melanogenesis. AB - The Tec cytoplasmic tyrosine kinase is a member of a family of src-like proteins that are thought to play important roles in hematopoiesis. Here we describe the temporal and spatial expression of the Tec gene during embryogenesis and in the adult. Our data demonstrate that embryonic Tec expression is restricted to distinct hematopoietic cells as well as structures and cell types that share a common feature of containing fluid in an enclosed cavity, e.g., endothelial cells. In addition, Tec is expressed in melanocytes late in gestation. The observed developmental expression pattern of Tec suggests a role for this gene in several aspects of hematopoiesis and/or blood vessel development as well as in late stages of melanogenesis. PMID- 9419412 TI - Regulation of P-glycoprotein expression in cyclic AMP-dependent protein kinase mutants. AB - Multidrug resistance (MDR) in cancer poses a major obstacle to the success of chemotherapy. We previously reported that cyclic AMP (cAMP)-resistant mutants of the Chinese hamster ovary and the mouse adrenal cortical carcinoma cells harboring defective regulatory (RI alpha) subunits of the cAMP-dependent protein kinase (PKA) are more sensitive than wild-type cells to chemotherapeutic agents that are substrates for P-glycoprotein. In addition, a transfectant overexpressing a mutant RI alpha cDNA showed similar increased sensitivity to these drugs. The altered drug sensitivity in the RI alpha mutants results from reduced expression of the mdr gene, suggesting that PKA may regulate its expression. In this study, we evaluated the sensitivity of several Chinese hamster ovary catalytic (C) subunit mutants to various anticancer drugs. Like the RI alpha subunit mutant, the C subunit mutants also exhibit decreased kinase activity and unresponsiveness to growth inhibition by cAMP. However, in contrast to the RI alpha subunit mutant, the C subunit mutants are not multidrug sensitive and maintain P-glycoprotein expression levels comparable to those of wild-type cells. Furthermore, the C subunit mutants display the same resistance patterns as wild-type cells to P-glycoprotein substrates, including Adriamycin, Taxol, and colchicine. No significant difference was observed in their sensitivity to non MDR drugs, such as 5-fluorodeoxyuridine, between wild-type, RI alpha, and C subunit mutant cells. These results suggest that the increased multidrug sensitivity in the PKA mutant cells results from alteration of the RI alpha subunit and not the kinase activity, thus implying novel functions for the RI alpha subunit. Therefore, genetic alteration of the RI alpha subunit of PKA may modulate drug resistance in cancer. PMID- 9419414 TI - Cripto-1 inhibits beta-casein expression in mammary epithelial cells through a p21ras-and phosphatidylinositol 3'-kinase-dependent pathway. AB - Cripto-1 (CR-1) is a recently discovered protein of the epidermal growth factor family that does not directly activate any of the known erbB type 1 tyrosine kinase receptors. Also, CR-1 stimulates the growth of HC-11 mouse mammary epithelial cells. We found that prior treatment of HC-11 cells with exogenous CR 1 induced a competency response to the lactogenic hormones dexamethasone, insulin, and prolactin (DIP) with respect to the induction of the milk protein beta-casein. In contrast, simultaneous treatment of mouse HC-11 cells with CR-1 in the presence of DIP inhibited beta-casein expression. The inhibitory effects of CR-1 on beta-casein expression in response to DIP were not unique to this mouse mammary epithelial cell line, because beta-casein and whey acidic protein expression in primary mouse mammary explant cultures established from midpregnant mice were also differentially inhibited by several epidermal growth factor related peptides including CR-1. The mitogenic and differentiation effects of CR 1 are mediated by the binding of CR-1 to a cell surface receptor that is known to activate the ras/raf/mitogen-activated protein kinase (MAPK)/MAPK kinase pathway. The inhibitory response of CR-1 in HC-11 cells on beta-casein expression after treatment with DIP can be attenuated by B581, a peptidomimetic farnesyltransferase inhibitor that blocks p21ras farnesylation and activation, and by the phosphatidylinositol 3'-kinase (PI3k) inhibitor LY 294002 but not by PD 98059, a MAPK kinase inhibitor that blocks MAPK activation. These data suggest that the ability of CR-1 to block lactogenic hormone-induced expression of beta casein is mediated through a p21ras-dependent, PI3k-mediated pathway. This is further substantiated by the observation that CR-1 is able to stimulate the tyrosine phosphorylation of the p85 PI3k regulatory subunit and to increase the activity of PI3k in HC-11 cells. PMID- 9419415 TI - Constitutive activation of Stat3 in fibroblasts transformed by diverse oncoproteins and in breast carcinoma cells. AB - Signal transducers and activators of transcription (STATs) were originally identified as key components of signaling pathways involved in mediating responses to IFNs. Previous studies showed that the Src oncoprotein constitutively activates one STAT family member, Stat3. In this study, we investigated STAT activation in a panel of rodent fibroblast cell lines stably transformed by diverse viral oncoproteins. Using a temperature-sensitive mutant of v-Src, we determined that Stat3 is activated within 15 min of shift from nonpermissive to permissive temperature for cell transformation. This finding indicates that v-Src tyrosine kinase activity is required for Stat3 activation and suggests that Stat3 is proximal to signaling initiated by Src. In addition, Stat3 activation is induced by another nonreceptor tyrosine kinase, v-Fps; by polyoma virus middle T antigen, which activates Src family kinases; and by v-Sis, which acts as a ligand for the platelet-derived growth factor receptor. In contrast SV40 large T antigen, which transforms cells through different mechanisms, and the v-Ras and v-Raf oncoproteins, which lie in signaling pathways downstream of tyrosine kinases, do not activate Stat3. We did not detect significant activation of Stat1, Stat5, or Stat6 in fibroblasts transformed by the viral oncoproteins investigated. Moreover, Stat3 is activated in response to epidermal growth factor (EGF) but not heregulins in immortalized normal human breast epithelial cells. Because constitutive activation of c-Src and EGF receptor kinases is associated with the progression of breast cancer, we examined activation of STATs in human cell lines derived from breast carcinomas. We detected constitutive activation of Stat3 in five of nine breast carcinoma cell lines but not in normal breast epithelial cells. Furthermore, experiments with an EGF receptor-specific inhibitor indicated that the constitutive activation of Stat3 in these breast carcinoma cell lines is not necessarily dependent on signaling through the EGF receptor, although EGF stimulation further increases Stat3 activation. Taken together, our results demonstrate that selective activation of Stat3 is a common event during oncogenic transformation that directly or indirectly involves activation of specific tyrosine kinase signaling pathways. PMID- 9419416 TI - Differential regulation of the pocket domains of the retinoblastoma family proteins by the HPV16 E7 oncoprotein. AB - The human papillomavirus E7 oncoprotein binds to the retinoblastoma (Rb) tumor suppressor protein, and the binding to Rb correlates with the oncogenic potential of E7. Recent studies from several laboratories indicated that the half-life of the Rb protein is reduced in cells that are stably transformed with E7, suggesting that E7 could induce the proteolytic degradation of Rb. To investigate whether the Rb degradation is a primary effect of E7 or a result of altered cell phenotype, we sought to develop assays that can distinguish between the two possibilities. Using recombinant adenovirus expressing the human papillomavirus type 16 E7 protein, we show that the expression of E7 leads to an increased rate of decay of the Rb protein. Moreover, Rb degradation immediately follows the expression of E7 suggesting that it is an early and primary effect. Consistent with a previous study, we observed that the E7-induced degradation of Rb can be blocked by the inhibitors of the 26S proteasome. We have also developed a transient transfection assay for the E7-induced degradation of Rb. Using this assay, we show that the pocket domain of Rb is necessary and sufficient for the E7-induced degradation. However, the proteolysis is relatively specific for Rb because the level of p107 or p130 was not significantly altered by the expression of E7. Thus, although E7 binds to all three members of the Rb family of proteins, the proteolysis is much more efficient in the case of Rb. In the transient transfection assays, adenovirus E1A and SV40 large T antigen failed to induce degradation of Rb, suggesting that the Rb degradation is a unique property of the E7 oncoprotein. PMID- 9419417 TI - Overexpression of normal c-Src in poorly metastatic human colon cancer cells enhances primary tumor growth but not metastatic potential. AB - Whereas genetic paradigms are now defined for the development of human colon cancer, little is known regarding the mechanisms that regulate development of the metastatic phenotype. Recent reports have indirectly linked the expression and activation of c-Src to the process of human colon cancer metastasis. Whereas v Src, a highly activated mutational derivative of c-Src, has been shown to induce metastasis, normal c-Src has not been tested for this property. We hypothesized that c-Src overexpression in the milieu of a poorly metastatic cancer cell might permit the development of a highly metastatic cell. Two poorly metastatic human colon cancer cell lines were stably transfected with expression vectors encoding normal human c-Src. Clones producing 4-10-fold more c-Src than controls were injected s.c. and intrasplenically into the nude mouse to assess primary tumor growth and liver metastatic potential. Whereas metastatic potential was unaffected, primary tumor growth in vivo was significantly enhanced by c-Src overexpression. No effects on rates of tumor cell proliferation were seen in vitro. Our findings suggest that normal c-Src may be necessary but is insufficient for the induction of the metastatic phenotype. PMID- 9419418 TI - Regulation of the laminin beta 1 (LAMB1), retinoic acid receptor beta, and bone morphogenetic protein 2 genes in mutant F9 teratocarcinoma cell lines partially deficient in cyclic AMP-dependent protein kinase activity. AB - We stably transfected a gene encoding a dominant negative regulatory subunit of cyclic AMP (cAMP)-dependent protein kinase A (PKA) into F9 cells and generated cell lines partially deficient in PKA activity (DN16 and DN19). In these cell lines, the retinoic acid (RA) receptor beta and laminin beta(1) chain (LAMB1) genes were regulated normally by RA alone, indicating that in the absence of exogenous modulation of cAMP levels, the PKA signaling pathway does not seem to play a major role in the RA-associated regulation of these genes. However, alterations in gene regulation were observed when the mutant cell lines were treated with a combination of RA and cAMP analogues. Moreover, in the DN16 cell line, which exhibits the lowest PKA activity among the mutant cell lines [22% of wild type (WT) at 1 microM cAMP], there was a significant decrease in the cAMP associated activation of the LAMB1 gene DNase I hypersensitivity site 2 enhancer, as measured by chloramphenicol acetyl transferase assays. Using electrophoretic mobility shift assays, less protein binding was observed at one of the motifs (C2) within this enhancer region in the DN16 cells as compared to the F9 WT cells after treatment of the cells with RA and cAMP analogues for 24 h. Furthermore, no increase in C2 binding was observed when extracts from RA-treated F9 ST or DN16 cells were subjected to in vitro phosphorylation, suggesting that PKA is involved in the induction of the C2-binding protein in RA-treated cells. In contrast to the results with RA receptor beta and LAMB1, the effects of cAMP analogues on the RA-associated regulation of the bone morphogenetic protein 2 gene were not altered in the cell lines that exhibited reduced PKA activity. These results suggest that a partial reduction in PKA activity is not sufficient to abrogate the effects of cAMP analogues on all of the genes regulated by RA. PMID- 9419419 TI - Redundant functions of B-Myb and c-Myb in differentiating myeloid cells. AB - We show in this report that the human myeloid leukemia cell line GFD8 is a useful model to compare the biological function of the structurally related c-Myb and B Myb proto-oncogenes and to investigate the c-myb domains required for this function. GFD8 cells are dependent for growth on granulocyte-macrophage colony stimulating factor and differentiate in response to phorbol myristate acetate (PMA). We have stably transfected this cell line with constructs constitutively expressing c-Myb or B-Myb. Deregulated expression of both c-Myb and B-Myb inhibited the differentiation observed in response to PMA and, in particular, the induction of the CD11b and CD11c antigens on the cell surface, and the induction of adherence. Furthermore, c-Myb and B-Myb enhanced expression of CD13 upon PMA treatment. Although deregulated Myb expression did not alter the growth factor dependence of the cells, it led to an increase in G2 relative to G1 arrest in cells induced to differentiate in response to PMA, whereas control vector transfected cells were blocked mostly in G1. This decrease in G1 block took place despite normal induction of the cyclin-dependent kinase inhibitor protein p21 (CIP1/WAF1). Thus, GFD8 cells stably expressing the human B-Myb protein behaved in a manner indistinguishable from those stably expressing C-Myb for both differentiation and cell cycle parameters. In agreement with these findings and differently from most previous reports, transactivation assays show that B-myb can indeed act as a strong activator of transcription. Finally, we demonstrated that although the DNA-binding domain of c-myb is required for both the differentiation block and the shift in cell cycle after PMA treatment, phosphorylation by casein kinase II and mitogen-activated protein kinase at positions 11 and 12 or 532 of c-myb, respectively, are not. We conclude that c Myb and B-Myb may activate a common cellular program in the GFD8 cell line involved in both differentiation and cell cycle control. PMID- 9419420 TI - Human erythropoietin receptor increases GATA-2 and Bcl-xL by a protein kinase C dependent pathway in human erythropoietin-dependent cell line AS-E2. AB - Erythropoietin (Epo) is a cytokine known to stimulate proliferation and differentiation of erythroid cells. However, recent gene disruption experiments demonstrated that Epo receptor signaling is not an obligatory step in erythroid differentiation. Here, we describe the role of Epo in proliferation, terminal differentiation, and apoptosis in a novel human Epo-dependent cell line, AS-E2. Upon withdrawal of Epo, the cells ceased to proliferate and underwent apoptotic death. Accompanying this cell death, an increase in the number of hemoglobin positive cells of approximately 2-fold was observed. This was associated with immediate up-regulation of the GATA-1:GATA-2 ratio and down-regulation of Bcl-xL. Treatment with Epo or 12-O-tetradecanoyl-phorbol-13-acetate (TPA) up-regulated expression of GATA-2 and Bcl-xL, and these elevations were inhibited by inhibitors of protein kinase C (PKC), H7 and H8. HA1004, a structural analogue of H7 but a poor inhibitor of PKC, had no inhibitory effect. Therefore, in AS-E2 cells, it is likely that Epo plays a role in (a) proliferation, (b) inhibition of differentiation, and (c) survival, by maintaining GATA-2 and Bcl-xL expression through activation of PKC. PMID- 9419421 TI - Positive and negative regulation of the human thymidine kinase promoter mediated by CCAAT binding transcription factors NF-Y/CBF, dbpA, and CDP/cut. AB - The proximal CCAAT element located 38 bp upstream of the transcription initiation site contributes to the human thymidine kinase (htk) promoter activity, because site-directed mutagenesis of a 10-bp region containing this CCAAT motif (TKC1) reduced the promoter activity by 55%. Through binding site competitions and antigenic cross-reactivity, the major factor that binds TKC1 from both HeLa and hamster nuclear extracts is identified as NF-Y/CBF. In serum-stimulated cells, the binding of NF-Y/CBF to TKC1 increased gradually, reaching a plateau at the S phase. In cell transfection assays, a dominant-negative mutant of NF-Y/CBF inhibited the htk promoter in a dosage-dependent manner, providing direct evidence that NF-Y/CBF is required for maximal htk promoter activity. Recently, it has been demonstrated that the site occupied by NF-Y/CBF also binds the serum inducible dbpA and dpbB. We show here that recombinant dbpA interacts with the htk promoter, and overexpression of dbpA can stimulate htk promoter activity mediated through TCK1. In contrast, CDP/cut, the CCAAT displacement protein with known repressor property, binds the htk promoter through both the proximal and distal CCAAT elements. Our discovery that CDP/cut binds the htk promoter primarily in quiescent cells and that overexpression of CDP/cut inhibits htk promoter activity provides an explanation for the reported dramatic increase in htk promoter activity in serum-starved cells when both CCAAT elements were mutated. Thus, a combination of suppression in quiescent cells and activation in serum-stimulated cells mediated through various CCAAT-binding proteins may account in part for the induction of htk promoter activity as quiescent cells reenter the cell cycle. PMID- 9419422 TI - Differential cation regulation of the alpha 5 beta 1 integrin-mediated adhesion of leukemic cells to the central cell-binding domain of fibronectin. AB - Normal and neoplastic leukocytes interact with the central cell-binding and carboxyl-terminal regions of fibronectin (FN) primarily via the alpha(4) beta(1) and alpha(5) beta(1) integrins. By using a unique centrifugation-based cell adhesion assay and affinity chromatography of the cell surface-labeled integrins, we show in this study that the constitutive alpha(5) beta(1)-dependent attachment of three leukemic cell lines, BV-173, K562, and Nalm-6, to FN or to a 110-kDa central cell-binding fragment of FN was totally inhibited at 37 degrees C by preincubation of the substrate with either antibodies to the arginine-glycine aspartic acid-containing region (3Fn-9 module) or to the synergistic region (3Fn 9 module) of FN. Similar results were obtained when assays were carried out at 4 degrees C, suggesting that energy-dependent events were not involved. On the other hand, only the antibody against the 3Fn-10 module was able to detach most firmly adherent cells. Constitutive cell attachment to the 110-kDa fragment was cation dependent, with the order of efficacy of the cation being Mn2+ > Mg2+ > Ca2+, and Ca2+ was only effective for BV-173 cells. Antibodies against the alpha(5) beta(1) integrin or the alpha(5) subunit completely impaired cell attachment of all cell lines, whereas several blocking anti-beta(1) subunit antibodies, including 4B4, P4C10, and AIIB2, differentially perturbed cell adhesion of BV-173, depending on which cation was present. These anti-beta(1) blocking antibodies, whose epitopes map to a region distant from the one expected to contain the beta(1) putative cation binding sites, seemed to lock the higher activation state of this integrin attained in the presence of Mg2+ and to preserve it during the subsequent adhesion events irrespectively of the presence of the low avidity state-inducing Ca2+ ion. Because the higher binding avidity displayed by BV-173 could not be explained by a higher degree of preclustering of alpha(5) beta(1) integrin in this cell line compared to K562, these findings suggest that cations might act as allosteric activators of integrin function. Whether this novel cation-dependent parameter of alpha(5) beta(1) integrin-FN interaction might contribute to the growth control and/or tissue dissemination of leukemic cells remains to be determined. PMID- 9419423 TI - Transformation by Wnt family proteins correlates with regulation of beta-catenin. AB - Several members of the Wnt family of secreted factors are strongly implicated as regulators of mammary cell growth and differentiation. To investigate Wnt signaling in mammary cells, we have assessed the abilities of 10 different Wnt genes to cause transformation of C57MG mammary epithelial cells and in parallel studied their effects on beta-catenin, a component of the Wnt-1 signaling pathway. Autocrine transforming potential was tested by expression of Wnt proteins in C57MG cells, and paracrine effects were evaluated by coculture of C57MG cells with fibroblasts secreting different Wnt proteins. Western blotting confirmed the expression of each Wnt protein in the relevant cell lines. Activities of the 10 Wnts tested were divisible into three groups. Wnt-1, Wnt-2, Wnt-3, and Wnt3a induced strong transformation and an elongated refractile cell morphology. Wnt-6 and Wnt-7a produced weak morphological changes. Wnt-4, Wnt-5a, Wnt-5b, and Wnt-7b had no effect at all on C57MG morphology. Analysis of beta catenin levels showed that the transforming Wnts induced accumulation of cytosolic beta-catenin, whereas nontransforming Wnts did not. These result demonstrate that several Wnt family members are capable of elevating beta-catenin levels and suggest that their signaling pathways share intracellular signaling components. The correlation between increased cytosolic beta-catenin levels and C57MG transformation supports a role for beta-catenin in transformation of these cells. These data also imply the existence of receptors that respond to certain Wnt proteins but not to others. PMID- 9419424 TI - The biochemical status of the DNA synthesome can distinguish between permanent and temporary cell growth arrest. AB - We previously identified and characterized the human leukemia (HL-60) cell DNA synthetic machinery as a multiprotein form of DNA polymerase, which was designated the DNA synthesome. This multiprotein replication complex contains DNA polymerases alpha and delta, primase, replication factor C, replication protein A, helicase, poly(ADPribose) polymerase, proliferating cell nuclear antigen, DNA ligase I, and topoisomerases I and II. Recently, the HeLa cell-derived DNA synthesome was identified as a discrete high molecular weight protein band in native polyacrylamide gels. Here, we report our findings regarding the change in the organizational status of the DNA synthesome when HL-60 cells undergo either terminal differentiation or temporary G1 growth arrest. We observed that the HL 60 cell DNA synthesome also migrates as a discrete high molecular weight protein band in nondenaturing polyacrylamide gels. This high molecular weight protein band was present in nuclei derived from both actively cycling cells and aphidicolin-arrested cells but was absent in TPA-induced terminally differentiated cells. We also found that DNA polymerase delta, replication factor C, and proliferating cell nuclear antigen are absent in cells that are induced to differentiate in response to 12-O-tetradecanoyl phorbol-13-acetate treatment but are present in actively cycling cells. The level of replication protein A in differentiated cells was similar to that of cycling cells, whereas the level of annexin I, a cytoskeleton protein, is higher in differentiated cells than it is in actively cycling cells. We conclude that the DNA synthesome remains integrated and inactive in temporarily growth-arrested cells but is disassembled in differentiated cells. Furthermore, we conclude that disassembly of the organized replication complex is a specific cellular event in the process of permanent cell cycle exit and that the process leading to disassembly may be regulated, in part, at the level of gene transcription. PMID- 9419425 TI - Pim-1 protein expression is regulated by its 5'-untranslated region and translation initiation factor elF-4E. AB - Expression of Pim-1, an oncogenic serine/threonine kinase, is highly regulated at the transcriptional, posttranscriptional, and posttranslational levels. Here, we report that expression of Pim-1 kinase is additionally regulated at the translational level. Pim-1 protein expression did not increase in Hut-78 lymphocytes in response to PMA1/ionomycin stimulation despite approximately 20 fold increases in mRNA levels, suggesting that translation was repressed. Sequence analysis of the 5'-untranslated region (UTR) indicated a long (400 nucleotide), 76% G + C-rich region, characteristics known to inhibit translation. Deletion of the 5'-UTR of pim-1 increased translation of the Pim-1 protein approximately 10-fold in vitro in reticulocyte lysates and approximately 1.6-fold in vivo in NIH-3T3 cells. When full-length 5'-UTR-containing pim-1 cDNA constructs were transfected into NIH-3T3 cells overexpressing eukaryotic translation initiation factor 4E (eIF-4E), approximately 6-fold higher levels of Pim-1 protein were produced, as compared to that produced in control NIH-3T3 cells. Moreover, eIF-4E overexpression had little effect in the absence of the 5' UTR, suggesting that it relieved 5'-UTR-mediated inhibition of Pim-1 expression. PMID- 9419426 TI - The discovery and use of HLA-associated epitopes as drugs. AB - MHC receptors "display" peptide fragments to T cells. These peptides are predominantly derived from proteins expressed within or ingested by the presenting cell. Since empty MHC molecules are highly unstable, peptide ligands are bound prior to MHC surface expression and the ensuing t1/2 off rates are often on the order of days. It is the remarkable stability of MHC/peptide complexes, which provide us an opportunity to purify MHC molecules from infected, transfected, or antigen pulsed cells and subsequently identify the naturally processed peptides being presented. On the other hand, the stability of MHC/peptide complexes substantially reduces the potency of parenterally administered peptides in vivo. Using serial immuno-affinity chromatography and mass spectrometry, naturally processed peptides can be identified. When these peptides are then encoded into nucleic acid and delivered parenterally, they are highly immunogenic. Application of these techniques to induce vigorous CTL responses will be discussed. PMID- 9419427 TI - Influence of the affinity of selecting ligands on T cell positive and negative selection and the functional maturity of the positively selected T cells. AB - Interaction of the TCR on immature thymocytes with ligands on antigen presenting cells can lead to different fates including positive and negative selection. The affinity of the selecting ligands plays an important role in determining these outcomes. We used the 2C TCR transgenic model to evaluate the efficacy of ligands with widely differing affinity (3 x 10(3) - 2 x 10(6) M-1) for the 2C TCR in mediating thymic negative and positive selection. Our results support the conclusions that the deletion of immature thymocytes is not only mediated by high affinity ligands but also by low-affinity/avidity ligands. However, high- and low affinity ligands differ in their requirements for negative selection. We also present evidence that positive selection is not an all or none process but depending on the strength of interaction between the ligand and the TCR during the positive selection process can result in single positive thymocytes that are at different stages of functional maturity. PMID- 9419428 TI - Antigen-presenting cells and the selection of immunodominant epitopes. AB - Cellular immune responses are directed against a narrow set of immunodominant peptides derived from complex antigens. By contrast, epitopes that are hidden or infrequent targets of immune responses have been termed cryptic or subdominant. Although the identification of immunodominant epitopes is important for vaccine development, understanding immunological reactivity to cryptic or subdominant epitopes may hold clues to autoimmunity. We have examined the role of antigen presenting (APC) cells in the selection of class Ii-restricted epitopes for display to T lymphocytes. The formation and MHC-restricted presentation of distinct antigenic epitopes is directly dependent upon processing and ligand binding reactions within APC. A novel MHC heterodimer, HLA-DM, facilitates the binding and presentation of peptides by class II DR, DP, and DQ molecules. We have demonstrated that some epitopes derived from endogenous antigens bind class II proteins independent of DM, whereas the presentation of other endogenous peptides is greatly influenced by DM expression. Targeting of exogenous antigens into specialized processing compartments within APC appears to overcome the requirement for DM in antigen presentation. These studies suggest HLA-DM may play a role in epitope selection and immunodominance. PMID- 9419429 TI - Functional and structural issues related to epitope cross-recognition by T cells. AB - T cell epitope recognition is a pivotal process in the immune response. Structural and functional data obtained in recent years have contributed greatly to an understanding of how epitopes are formed by MHC/peptide complexes and how such epitopes are engaged by T cell receptors. Emerging as well from these studies is an appreciation of the tension that exists between epitope specificity and degeneracy, stemming from the highly flexible nature of ligand engagement by T cell receptors. The issues surrounding epitope cross-reactions are particularly germane to understanding the role played by epitope mimicry in contributing to autoreactive responses and autoimmune pathology. PMID- 9419430 TI - Agonistic activity of a CD40-specific single-chain Fv constructed from the variable regions of mAb G28-5. AB - A single-chain Fv (sFv) was expressed from the variable regions of the CD40 specific mAb G28-5. The molecule bound CD40 with a high affinity (2.2 nM) and was a monomer in solution. Surprisingly, G28-5 sFv was a potent CD40 agonist that rapidly crosslinked CD40 on the cell surface but did not crosslink CD40-Ig in solution. G28-5 sFv was a more potent agonist than G28-5 IgG and was able to stimulate CD40 responses by B cells and monocytes. G28-5 IgG partially blocked, whereas G28-5 sFv augmented CD40 responses during stimulation with natural ligand (gp39-CD8 fusion protein). These results indicate that the functional activity of ligands built from the binding site of G28-5 is highly dependent upon the size and physical properties of the molecule both in solution and on the cell surfaces. PMID- 9419431 TI - Immune surveillance: paraneoplastic or environmental triggers of autoimmunity. AB - Autoimmunity associated with tumor cell development seems an important mechanism by which to prevent progression to clinical cancer. In this brief review, tumor autoantigens associated with paraneoplastic syndrome, non-HLA-associated organ specific autoimmune diseases, and the highly cell-specific autoimmune eradication of the islet beta cells in type 1 diabetes are compared and discussed. It is suggested that autoreactivity is important in preventing tumor formation; however, it may be at the expense of the development of autoimmune disease. Although the cytotoxic T lymphocytes (CTL) induction by HLA class I has been studied and used in clinical trials, little is understood about the initiation and HLA class II mediated induction of an immune response to neoplastic cells. This induction apparently takes place because paraneoplastic disorders are often due to an immune response to the tumor cell resulting in a cross-reactivity with a normally expressed autoantigen on a remote nontumor-associated target cell. The problem of immune surveillance to eradicate neoplasm or downregulate pathological autoimmunity are therefore closely related phenomena. An improved understanding of immune mediated tumor suppression should therefore greatly benefit immunotherapy of type 1 diabetes, and the two areas of research would benefit from an interdisciplinary endeavor. PMID- 9419432 TI - T cell recognition of self and altered self antigens. AB - T lymphocytes bearing alpha/beta TCR recognize antigens in the context of self MHC molecules, and this recognition leads to growth, differentiation, and effector functions. Recently, it has become clear that altered peptides generated by single amino acid substitution of the antigenic peptide can alter the patterns of differentiation and effector functions of the responding T lymphocytes. By defining the pattern of recognition and residues of the cognate ligand that bind to the TCR, altered peptide ligands (APLs) have been generated by selectively substituting the TCR contact residues in the antigenic peptide. These APLs have been utilized in vitro to study the biology of T cell function and alterations in the T cell signaling pathway. In vivo APLs have been utilized to study the mechanism of positive selection in the thymus and in regulation of autoimmune diseases. With this basic knowledge, APLs that can either hypo- or hyper stimulate T cell function can be generated that can specifically alter (inhibit or enhance) immune responses in vivo in autoimmune diseases and cancers. PMID- 9419433 TI - Role of MHC class I molecules in autoimmune disease. AB - The MHC class I molecules play a pivotal role in triggering cellular immune responses, binding and presenting intracellularly derived peptide antigens. Studies of MHC class I expression revealed a complex regulatory mechanism that integrates tissue-specific and hormonal modulation. Dynamic regulation occurs in the thyroid, in response to hormonal repression by TSH and stimulation by thyroid hormone. This dynamic cycle provides the basis for proposing the model that such regulation is important to maintain tolerance to self-antigens in tissues synthesizing large amounts of secretory proteins. Failure to appropriately regulate class I levels is predicted to result in autoimmunity. In support of this model, we found that class I-deficient mice are resistant to the experimentally induced autoimmune diseases, SLE, and blepharitis. Furthermore, pharmacological treatment with an agent that reduces class I expression also reduces the incidence and severity of both experimental and spontaneous autoimmune SLE. PMID- 9419434 TI - Initiation and regulation of CNS autoimmunity. AB - Our studies addressed the questions of how self-reactive T cells escape tolerance and what stimuli cause these T cells to initiate autoimmune responses. We employed experimental allergic encephalomyelitis (EAE) as an animal model of multiple sclerosis (MS). Endogenous expression of myelin basic protein (MBP) induces tolerance in T cells that recognize one region of MBP, whereas T cells specific for a different region escape tolerance. Triggers of disease induction were investigated in a T-cell receptor (TCR) transgenic model in which the majority of T cells recognize the MBP epitope that does not induce tolerance. EAE occurs spontaneously in this model and the incidence of disease depends on microbial exposure. EAE can also be actively induced by immunization with MBP peptide accompanied by injection of pertussis toxin as well as by administration of pertussis toxin alone. Immunization with MBP peptide without pertussis toxin, however, stimulates the transgenic T cells, but the activated T cells do not accumulate in the central nervous system (CNS) or induce EAE. Our studies suggest that initiation of autoimmune disease involves complex interactions between the neuroendocrine system as well as the innate and specific immune systems. PMID- 9419435 TI - Autoimmune responses against acetylcholine receptor: T and B cell collaboration and manipulation by synthetic peptides. AB - Myasthenia gravis (MG) and experimental autoimmune MG (EAMG) are induced by antibodies (Abs) against self acetylcholine receptor (AChR). We have mapped the T and B cell epitopes on AChR alpha subunit in human MG and in EAMG-susceptible (C57BL/6, B6) and nonsusceptible mouse strains. A T-cell epitope within residues alpha 146-162 (P14) of Torpedo californica (t)AChR plays an important role in EAMG pathogenesis of the auto Ab-induced disease. P14-specific T cell (P14Th) lines from tAChR-primed B6 mice activated, in vivo and in vitro, tAChR-primed B cells that secreted anti-AChR Abs directed against four other regions on the tAChR alpha-chain, but not against P14 itself. P14Th cells are pathogenic because they help B cells that make Abs against a conserved tAChR region (t alpha 122 150) involved in ACh binding. These Abs cross-react with region alpha 122-150 of mouse (m)AChR, thereby disrupting its normal physiological function. Thus, a T cell epitope not recognized by Abs plays an active role in B cell responses against other epitopes on the protein. We have found that in B6, the MHC region 62-76 of I-A beta(b) is involved in the presentation of P14 to T cells. Anti peptide Abs, prepared in BALB/c, were found to inhibit in vitro the proliferation of P14-specific T-cells. Furthermore, this MHC peptide elicited Abs in B6 mice and we are investigating whether immunization of B6 with this peptide, before priming with tAChR, would suppress in vivo the T-cell response to the epitope in P14. Thus, these preliminary results would suggest that immunization with the MHC peptide might be employed for control of the autoimmune disease. PMID- 9419436 TI - Binding of human IgG myeloma proteins to autologous T-cell receptor determinants. AB - IgG myeloma proteins (MPs) produced by monoclonal plasma cells derived from B2 lymphocytes have been reported to bind to various autoantigens but the binding generally has been of low affinity. Moreover, T cells from some multiple myeloma patients can respond specifically to idiotypes of their own paraproteins. We analyzed the capacity of more than 20 human IgG MP to bind, a recombinant single chain molecule containing complete V beta 8.1 and V alpha 1 structures, sets of synthetic peptide epitopes corresponding to a complete TCR beta chain, and a set of CDR1 epitopes corresponding to 24 human V beta gene products, and intact monoclonal T cells. Two of 20 MPs bound strongly to the recombinant TCR. Five of the same set, including these, bound to a synthetic epitope corresponding to the CDR1 segment. On a mass basis, the binding was approximately 1000-fold greater than that of pooled polyclonal IgG. The binding activity was confined to the Fab fragment and was specifically inhibitable by appropriate peptide determinants. Spectrotypic analysis using a set of CDR1 epitopes indicated that individual proteins showed characteristic binding patterns ranging from highly specific to relatively promiscuous. Highly reactive MPs also bound to TCR on intact cells in immunocytofluorescence by flow cytometry. These results are consistent with the relatively frequent occurrence of autoantibodies to TCR determinants and indicate that MPs can be derived from this autoantibody subset. PMID- 9419437 TI - T-cell receptor peptides as immunotherapy for autoimmune disease. AB - The observations in both mouse and rat models of experimental allergic encephalomyelitis (EAE) demonstrating restricted T-cell receptor (TCR) usage among pathogenic T cells has led to the generation of a new class of therapeutic vaccines composed of TCR V region peptides. Whether a similar approach will be of use in the treatment of human autoimmune disorders is still unclear. The experiments performed in our laboratory over the past several years have focused on two aspects of TCR peptide immunoregulation, namely, (1) how to identify the critical T-cell populations involved in the pathology of autoimmune disease, and (2) how to identify biologically relevant TCR peptides--those endogenous TCR peptides presented in association with MHC molecules on the surface of pathogenic T cells that are recognized by immunoregulatory T-cell populations. Results of our recently completed clinical studies regarding TCR V beta expression among CD4+ T cells in the cerebral spinal fluid (CSF) of patients with multiple sclerosis suggests that these cells may be an appropriate T-cell population to be targeted for TCR peptide therapy. In addition, our studies on the immune response to autologous, soluble TCR heterodimers may provide a strategy for the identification of new TCR peptide candidate vaccines. PMID- 9419438 TI - The role of iodine in autoimmune thyroiditis. AB - Like most cancers, autoimmune diseases generally are due to the interaction of a number of genetic traits with an environmental trigger. Autoimmune thyroiditis, a model of organ-specific autoimmune disease, is associated with iodine as a precipitating environmental factor. T cells from patients with chronic thyroiditis proliferate in response to normal human thyroglobulin, but fail to react with non-iodinated thyroglobulin. Using a selected monoclonal antibody, we were able to identify a binding site on thyroglobulin containing iodinated thyronine. The greatest affinity was for tetraiodothyronine and binding depended upon the number as well as the positions of iodines. We have also studied an inbred strain of mice, NOD-H2h4, that developed thyroiditis spontaneously. The onset of disease was hastened in a dose-dependent manner by adding iodine to the drinking water. The occurrence of disease was greater in conventional than in specific pathogen-free mice and correlated with T-cell proliferation and IgG2b antibody to thyroglobulin. PMID- 9419439 TI - Modulation of insulin-dependent diabetes mellitus (IDDM) in NOD mice by autoreactive T cells. AB - Insulin-dependent diabetes mellitus (IDDM) is a T-cell-mediated autoimmune disease characterized by the destruction of insulin-producing beta cells in the islet of Langerhans. Islet autoantigen-specific T cells play a major role in the pathogenesis of the disease. Susceptibility loci for autoimmune diabetes such as the major histocompatability complex (MHC) may function by producing different repertoires of T cells, which could gain autoreactivity following activation, resulting in autoimmune disease. However, all the T cells infiltrating the islets are not destructive. A number of autoreactive T-cell lines capable of preventing development of IDDM have been isolated. Most of these cell lines are reactive to self I-Ag7. Presence of these regulatory T cells along with the effector cells in nonobese diabetic (NOD) mice suggests that IDDM may be a result of the imbalance of these two types of cells. Modulation of the immune response by inducing autoreactive regulatory T cells could be a way of treating autoimmune disorders. PMID- 9419440 TI - Treatment of autoimmune diseases through manipulation of antigen presentation. AB - In rheumatoid arthritis, HLA-DR alleles associated with elevated relative risk share a common sequence in the third hypervariable domain of the major histocompatibility complex class II molecule (MHC II). Immunization of mice with a peptide vaccine comprised of the appropriate MHC II sequence in adjuvant blocked the onset and reduced the relapse rate of experimental autoimmune encephalomyelitis (EAE). A phase I clinical trial testing a single injection of a third hypervariable domain peptide from the HLA-DRB1*0401 sequence in alum adjuvant showed that the vaccine is well tolerated and generates an anti-HLA-DR antibody response in approximately 25% of the treated patients. A phase II trial testing multiple boosts is in progress. In a more antigen-specific approach, solubilized MHC II molecules loaded with an autoantigenic peptide are injected intravenously to induce unresponsiveness by the binding of the T-cell receptor (TCR) in the absence of costimulation. Appropriate soluble MHC II:autoantigenic peptide complexes inhibit the recall antigen proliferative response of T clones or draining lymph node cells, and reduce the progression of EAE and experimental autoimmune myasthenia gravis (EAMG). A test of a soluble HLA-DR2:myelin basic protein (MBP) complex in multiple sclerosis is progressing in phase I. PMID- 9419441 TI - Cytokine- and costimulation-mediated therapy of IDDM. AB - T cells from NOD mice display an age-dependent, TCR-inducible proliferative hyporesponsiveness that may be causal to IDDM. Exogenous IL-4 completely restores this hyporesponsiveness in vitro and prevents IDDM in vivo when administered to NOD mice. We therefore tested the hypothesis that stimulation of a Th2 response by either IL-4 or CD28 costimulation may block progression to IDDM. Low-dose IL-4 treatment beginning at 2 weeks of age (pre-insulitis) protects NOD mice from insulitis, sialitis, and thyroiditis, indicating that IL-4 modulates T cell migration to these inflammatory sites. Cytokine secretion profiles of stimulated T cells and assays of intrapancreatic cytokine concentrations revealed that IL-4 treatment prevents IDDM by stabilizing a protective Th2-mediated environment in the thymus, spleen, and pancreatic islets. Whereas treatment of NOD mice with an anti-CD28 mAb between 2 to 4 weeks of age inhibits destructive insulitis and protects against IDDM by enhancing IL-4 production by T cells, anti-CD28 treatment between 5 to 7 weeks of age does not prevent IDDM. Simultaneous anti-IL 4 treatment abrogates the protective effect conferred by anti-CD28 treatment. Our data demonstrate that stimulation of a Th2-cell-enriched environment in the pancreas during the inductive phase of disease development blocks progression to IDDM in NOD mice. PMID- 9419443 TI - T cell immunity to tumor antigens. PMID- 9419442 TI - Regulation of the inflammatory response in animal models of multiple sclerosis by interleukin-12. AB - Interleukin 12 (IL-12), a novel heterodimeric protein produced primarily by antigen-presenting cells, serves as a key regulator of innate and adaptive immune responses. In addition to being a potent inducer of IFN-gamma, IL-12 is widely considered to be the principal cytokine that regulates the generation of Th1 type effector cells. As the successful induction of experimental autoimmune encephalomyelitis (EAE) is associated with a strong Th1 type cellular response, we have evaluated the role of IL-12 in regulating the pathogenesis of EAE in SJL/J mice and Lewis rats. In both settings, treatment with IL-12 was found to accelerate the onset and increase the severity and duration of clinical disease. More importantly, administration of IL-12 to Lewis rats that had recovered from primary disease was found to trigger clinical relapse. In all instances, IL-12 induced exacerbation was associated with a profound increase in iNOS positive macrophages within the perivascular lesions. Although IL-12-induced IFN-gamma does not appear to be required for exacerbation of disease, neutralizing antibodies against murine IL-12 delay the onset and reduce the severity of adoptively transferred EAE, indicating a role for endogenous IL-12 as regulator of disease. Based on the above findings, effective inhibition of IL-12 in vivo may have great therapeutic value in the treatment of MS and other Th1-associated inflammatory disorders. PMID- 9419444 TI - Identification of peptides for immunotherapy of cancer. It is worth the effort. AB - As the nature of the T cell immune response is defined by T cell receptor recognition of small protein fragments, referred to as peptides, the identification of peptides would lead us to understanding and directing the T cell-mediated immune response. Immunogenic peptides might be used for vaccination and activation of the immune reaction against cancer- and virus-infected cells. Additionally, the knowledge of immunogenic peptides was expected to lead to blocking of allergic reactions and autoimmune diseases. Based on these assumptions, the search for immunogenic peptides was started in mice and man in the mid-1980s. After a decade of peptide identification and testing in vitro and in vivo, this may be a proper time to evaluate the results from the peptide related work and determine the possible applications of this knowledge for the next decade. In this review we discuss the identification of peptides, their use in murine models, as well as clinical data from peptide vaccinations or therapies. Potential hazards and limitations of peptide use in immunotherapy and other possible applications for peptides or peptide motifs in immunotherapy are evaluated. PMID- 9419445 TI - p53-based immunotherapy of cancer. AB - Immunotherapy targeting p53 missense mutations, which occur in nearly half of all human tumors, is limited by several factors, including the constraints of antigen processing and presentation. Due to the accumulation of mutated p53 molecules in tumors expressing p53 mutations, an alternative approach would be to target wild type sequence, CTL-defined p53 epitopes. Obviously, the possibility of an autoimmune response is a major potential drawback to this therapy. Immunization of BALB/c mice with bone marrow-derived dendritic cells (DC) generated in the presence of GM-CSF/IL-4 and prepulsed with the H-2Kd-binding wild-type p53(232 240) peptide has been shown to induce anti-peptide CTL. These effectors were cross-reactive against sarcomas expressing p53 missense mutations outside of the p53(232-240) epitope, but not within it. Mitogen-activated splenocytes, which express elevated levels of p53, were not sensitive to these CTL. The p53 peptide pulsed DC-based vaccine was shown to be effective in inducing tumor rejection in immunization and therapy models in the absence of any observable deleterious effect on naive mice. The murine model has now been extended to include the use of genetically modified DC-based vaccines as well. PMID- 9419446 TI - HER-2/neu oncogenic protein: issues in vaccine development. AB - Vaccine studies using whole tumor cells or heterogeneous mixtures of tumor antigens provide intriguing evidence that cancer vaccines might be effective. Now it is possible to test vaccines composed of well-characterized proteins and peptides. Testing vaccine formulations composed of known and defined antigens will allow a more precise determination as to why vaccines work when they work, and why they do not work when they fail. The demonstration that human malignancy is immunogenic and the definition of human tumor antigens has set the stage for a new generation of cancer vaccines directly targeting immunogenic cancer-related peptides and proteins. Many newly defined tumor antigens are self proteins. As an example, screening existent immunity in human melanoma has identified responses to nonmutated self proteins: MAGE, MART, gp100, and tyrosinase. Tolerance to self antigens now emerges as a possible mechanism of tumor immune escape. A new puzzle has emerged for tumor immunologists to solve; how to harness immunity to "self" tumor antigens for cancer therapeutics. PMID- 9419447 TI - Strategies for tumor elimination by cytotoxic T lymphocytes. AB - Despite differences in their tissue of origin, many tumors share high level expression of certain tumor-associated proteins. Our laboratory has focused on the possibility of utilizing antigenic components of these proteins as a focus for T-cell immunotherapy of cancer. The advantage of targeting such commonly expressed proteins is the fact that such therapy could be of value in eliminating many different types of tumors. A potential barrier in the identification of T cell epitopes derived from these proteins and presented by tumor cells is the fact that these proteins are also expressed at low levels in some normal tissues, and therefore, self-tolerance may eliminate T cells that are capable of recognizing these epitopes with high avidity. We have utilized two different murine model systems to explore the extent to which self-tolerance may limit the immune response to a tumor-specific antigen. The first compared the ability of mice deficient in expression of murine p53 (p53 knock-out mice) and normal mice, to respond against several epitopes of the p53 protein. The second model compares the ability of conventional mice with transgenic mice that express the influenza hemagglutinin in the periphery to respond to a dominant antigenic peptide of this transgene product. In both models we have investigated the effect self-tolerance has on elimination of tumors expressing the toleragen. PMID- 9419449 TI - In vitro immunization and expansion of antigen-specific cytotoxic T lymphocytes for adoptive immunotherapy using peptide-pulsed dendritic cells. AB - The design of an effective procedure to sensitize and expand antigen-specific cytotoxic T lymphocytes (CTL) in vitro is essential for the development of effective adoptive cellular immunotherapy protocols for cancer. We have analyzed the capacity of tissue culture-derived dendritic cells (DC) to present specific peptide epitopes to CTL precursors. Our results demonstrate that peptide-pulsed DC were efficient in generating CTL responses specific for various viral and tumor epitopes. Furthermore, IL-7 and IL-10 potentiated the ability of the peptide-pulsed DC to trigger antigen-specific CTL responses. The CTL generated using this procedure efficiently recognized the naturally processed antigens and could be expanded approximately 100- to 1000-fold in tissue culture in 10 to 15 days without a loss of activity and specificity. The results and procedures described herein may facilitate the development of effective CTL-based adoptive immunotherapy for chronic viral diseases and cancer. PMID- 9419448 TI - Recognition of melanoma-derived antigens by CTL: possible mechanisms involved in down-regulating anti-tumor T-cell reactivity. AB - Several T cell-recognized epitopes presented by melanoma cells have been identified recently. Despite the large array of epitopes potentially available for clinical use, it is still unclear which of these antigens could be effective in mediating anti-tumor responses when used as a vaccine. Preliminary studies showed that immunization of melanoma patients with epitopes derived from proteins of the MAGE family may result in significant clinical regressions. However, no sign of systemic immunization could be observed in peripheral blood of treated patients. Conversely, significant immunization (detected as increased antigen specific CTL activity in peripheral blood) was obtained by vaccinating HLA-A2.1+ melanoma patients with the immunodominant epitope (residues 27-35) of the differentiation antigen MART-1, but this immunization was not accompanied by a significant clinical response. To implement immunotherapeuties capable of significantly impacting disease outcome, it is necessary to identify the potential mechanisms responsible for the failure of some antigens to mediate significant anti-tumor responses in vivo. In the case of the MART-1(27-35) epitope, we hypothesize that one of these mechanisms may be related to the existence of natural analogs of this peptide in other human normal proteins. PMID- 9419450 TI - Tumor-specific immune response: current in vitro analyses may not reflect the in vivo immune status. AB - Although our knowledge and understanding of tumor-specific cytotoxic T lymphocytes (CTL) have expanded considerably, the long-term work needed to assay CTL has precluded their analysis in large numbers of patients. Moreover, in vitro culture steps may introduce major biases. New approaches to identify tumor specific CTL clones would be helpful. As a means to describe the in situ immune status by T-cell repertoire analysis, we developed the Immunoscope approach, a PCR-based method that allows us to determine the spectra of CDR3 lengths of the TCR chains displayed by complex populations of T cells. We review here some of our data about melanoma. Tumor-infiltrating lymphocytes of a melanoma patient were analyzed by different means and melanoma-specific T-cell clones were derived. Two categories of tumor-specific CD8+ CTL clones were derived from the infiltrate of a tumor-proximal invaded lymph node. The majority of T-cell clones specifically lyse the autologous tumor cell lines and predominantly recognize the HLA-A2/MART-1(27-35) peptide complex. The in vivo representativity of such CTL was assessed by the immunoscope technology. Among three MART-1-specific clones, none was detectable in situ. The other kind of tumor-specific CD8+ CTL did not lyse autologous melanoma cell lines but lysed the "fresh" autologous tumor cells in a MHC class I dependent manner. The immunoscope approach revealed that one of the latter was detectable in situ among tumor-infiltrating lymphocytes although not among PBMC. These data indicate that melanoma-specific lymphocytes that could not have been selected through conventional screening procedures may be important in tumor rejection. Our results suggest that a better characterization of tumor specific immune responses will be important for the optimization of specific immunotherapy strategies and the long-term follow-up of patients. PMID- 9419451 TI - Dendritic cells, interleukin 12, and CD4+ lymphocytes in the initiation of class I-restricted reactivity to a tumor/self peptide. AB - Cell-mediated immunity involving CD8+ lymphocytes is effective in mediating rejection of murine mastocytoma cells bearing P815AB, a tumor-associated and self antigen showing similarity to tumor-specific shared antigens in humans. Although this antigen may act as an efficient target for class I-restricted responses in immunized mice, neither P815AB expressed on tumor cells nor a related synthetic nonapeptide will activate unprimed CD8+ cells for in vivo reactivity, measured by skin test assay. We review evidence showing that the failure of P815AB to initiate CD8+ cell reactivity may be due to defective recruitment of accessory and Th1-like cells to the afferent phase of the response initiated by transfer of mice with dendritic cells pulsed in vitro with the P815AB peptide. Although the copresence of a T helper peptide in dendritic cell priming in vitro with P815AB may compensate for the poor generation of accessory and Th1 cells in the adoptively transferred mice, recombinant IL-12 can replace the helper peptide in both effects. Effective priming to P815AB in vivo is achieved by either exposing dendritic cells to IL-12 prior to P815AB priming or administering the recombinant cytokine in vivo. Different approaches suggest that IL-12 may act both on accessory cells to improve presentation of previously undescribed class II restricted epitopes of P815AB and on CD4+ cells to improve recognition of such epitopes. In particular, at the CD4+ cell level, IL-12 apparently acts as an adjuvant and an inhibitor of anergy induction. These data offer useful information for developing vaccination strategies using dendritic cells and class I-restricted tumor peptides in humans. PMID- 9419452 TI - Induction of dendritic cells (DC) by Flt3 Ligand (FL) promotes the generation of tumor-specific immune responses in vivo. AB - Daily treatment of mice with human Flt3 ligand (FL) induces the production of large numbers of MHC-class II+, CD11c+, DEC205+ dendritic cells (DC) in both lymphoid and nonlymphoid tissues. Because DC play a pivotal role in the induction of immune responses, we evaluated the effects of FL in the augmentation of anti tumor immune responses in vivo. Treatment with daily subcutaneous injections of FL not only caused complete regression of tumors in a significant proportion of mice challenged with a syngeneic methylcholanthrene-induced fibrosarcoma, but also caused a significant decrease in the tumor growth rate in the remaining mice. The effects of FL were both dose and schedule dependent, induced regression of tumors established as much as 7 days prior to beginning cytokine treatment, and was mediated by both T-cell mediated and non-T cell-mediated mechanisms. Histopathologic evaluation of tumors during and after systemic treatment with FL revealed a dense infiltration of mononuclear and myeloid precursor cells into both the tumor and tissue surrounding the tumor that generally correlated with the degree of regression and resolved after rejection of the tumor. Collectively, the data suggest that FL may be an important cytokine in the generation of effective anti-tumor immune responses in vivo and the treatment of cancer in situ. PMID- 9419453 TI - Dendritic cell-based immunotherapy of prostate cancer. AB - The immunotherapy of cancer, based on eliciting or enhancing the body's own capacity to mount an effective antitumor response, has produced encouraging early results in the areas of melanoma and renal-cell carcinoma. Such treatments utilizing dendritic cells (DC), immune cells that are excellent antigen presenters, are especially promising. We performed a phase I clinical trial assessing the administration of autologous DC pulsed with HLA-A0201-specific prostate-specific membrane antigen (PSMA) for the treatment of 51 men with hormone-refractory prostate cancer. Participants were divided into five groups receiving four or five infusions of peptides alone (PSM-P1 or PSM-P2; group 1 and 2, respectively), autologous DC (group 3), or DC pulsed with PSM-P1 or P2 (group 4 and 5, respectively). No significant toxicity was observed. Immune reactivity against PSM-P2 was detected in HLA-A2+ patients infused with DC pulsed with PSM P1 or -P2 (group 4 and 5). An average decrease in PSA was observed only in group 5. Seven partial responders were identified based on NPCP criteria + PSA. The excellent tolerance of this treatment approach, as well as the enhanced cellular responses, decreased PSA levels, and partial clinical responses in some patients suggests that it holds great potential in prostate cancer therapy. PMID- 9419454 TI - Human leukemia-derived dendritic cells: ex-vivo development of specific antileukemic cytotoxicity. AB - The human myeloid leukemias are a diverse group of disorders characterized by massive clonal expansion of myeloid cells showing variable degrees of differentiation block. Leukemic dendritic cells were generated in culture from chronic myelogenous leukemia (CML). These were used to stimulate autologous T cells to develop leukemia-specific cytotoxicity. Available data suggest that the cells responsible for the cytolytic activity are at least in part CD8+ and HLA restricted in their function. Additional data suggest that some anti-CML cellular activity may be Fas mediated. T-cell receptor studies provide evidence for an oligoclonal response implying a recognition of a limited number of antigens. We have used culture techniques similar to those used for CML to study the ability of AML cells to differentiate toward dendritic cells. Four of five patients have shown acute leukemia-derived dendritic cells. This work offers an avenue for the development of novel strategies for the control of human myeloid leukemias. PMID- 9419455 TI - Immunotherapy of colon cancer using chimeric mAb 31.1. AB - We have produced two monoclonal antibodies specific for membranes of colon carcinoma cells that demonstrate minimal if any cross reactivity with normal colon tissue. The antigen of one of the two antibodies (mAb 33.28) is extremely immunogenic eliciting both cell mediated and humoral immunity. The monoclonal antibodies developed in in vitro studies suggested strong antibody dependent cell cytotoxicity (ADCC) for both antibodies, but somewhat stronger for the mAb 31.1. The murine version of mAb 31.1 produced approximately 90% tumor cell destruction in the same period of time. Nude mice with LS174T xenografts (human colon carcinoma) were challenged with 1 x 10(6) cells given subcutaneously in the thigh. By day 10, well animals were incapacitated by tumor growth. Among those animals receiving 400 micrograms of intraperitoneal chimeric 31.1 at day one, all were protected and remained free of disease. Those challenged with chimeric antibody and human effector cells at day 7 demonstrated regression of established tumor nodules among 80% of the animals so treated. Therapy of patients with extensive primary as well as metastatic colon cancer may be found to respond to the above form of therapy post surgery, when employed alone or in combination with an effective cytotoxic (chemotherapeutic) agent. PMID- 9419456 TI - Tumor cell recognition by lymphocytes: is the MHC always essential? AB - Phenotypic and functional analyses of tumor-infiltrating lymphocytes (TILs) used in clinical trials revealed that cells other than CTLs can have antitumor efficacy. This observation led us to search for mechanisms for tumor recognition by lymphocytes that utilize alternatives to surface structures of tumor cells, for example, MHC antigen complexes; the latter are generally believed to be the immunogenic platforms for CTLs. Therefore, as a possible source of immunostimulatory activity, we compared the ability of plasma membrane components of tumor cell lines with first secreted tumor cell components and then intracellular tumor components to act as mitogenic sources for human TIL lines. Surprisingly, the latter was found to be most potent, particularly Oncoimmunin-L, which is a 45-kDa protein with sequence similarity to members of the serpin family of proteins. This protein, which has at least a 31% sequence identity to human leukocyte elastase inhibitor and stimulates [3H]-thymidine incorporation into the DNA of human TILs, may be found in the cytosol of many tumor cells. Taken together with our earlier work in which a 36-kDa protein, also of tumor cytosolic origin, was shown to induce differentiation of myeloid cells, we propose soluble factors derived from tumor cells as a pathophysiological source of tumor immunogenicity. Moreover, detailed biochemical and biophysical characterization of tumor cell-immunocyte interactions will define the tumor immunoenvironment. PMID- 9419457 TI - Contrast nephrotoxicity: predictive value of urinary enzyme markers in a rat model. AB - RATIONALE AND OBJECTIVES: We tested whether urinary enzymes are an accurate and useful marker of renal damage in a rat model of contrast media nephrotoxicity. METHODS: Thirty rats were pretreated with a combination of salt depletion, indomethacin, and contrast material. Alanine aminopeptidase (AAP), gamma glutamyltranspeptidase (GGT), and N-acetyl-beta-D-glucosaminidase (NAG) were measured before and 24 hr after injection of contrast material. Enzyme concentrations were correlated with glomerular filtration rate (GFR) and histology. RESULTS: Decreasing GFR and histopathologic changes were found only in rats treated with all three variables. NAG levels increased from baseline for both diatrizoate meglumine- and ioversol-treated animals (from 83.9 +/- 48.6 to 145.5 +/- 55.4 and from 69.41 +/- 43.6 to 123.1 +/- 50.7, respectively; P < 0.05 from baseline for ioversol) and declined in other groups. GGT and AAP levels did not correlate well with structural and functional changes. CONCLUSIONS: In this model of contrast nephrotoxicity, NAG concentration appears to correlate with structural and functional changes associated with contrast media nephrotoxicity. However, the large range of baseline values makes this of dubious clinical use. AAP and GGT levels appear less helpful in detecting renal damage. PMID- 9419458 TI - Interobserver agreement using computed radiography in the adult intensive care unit. AB - RATIONALE AND OBJECTIVES: We determined whether computed radiographs in an adult intensive care unit produce greater agreement between readers in descriptions of radiologic findings and interval changes. METHODS: Fifty-six conventional radiographs and 56 computed radiographs were obtained on 28 patients. Computed and conventional radiographs were imaged simultaneously in a single exposure with the use of a sandwich cassette. The two sets of films per patient were taken less than 1 week apart. RESULTS: Four readers evaluated the chest films and judged interval changes with the use of standardized confidence scales. Overall exposure and film quality also were evaluated. Computed radiographs were rated better in overall quality. Tubes and lines were detected with comparable accuracy. Interobserver agreement in the evaluation of radiologic descriptors was generally greater with computed radiographs (intraclass correlation). No differences were noted in interobserver agreement in the judgment of interval changes. CONCLUSIONS: The diagnostic usefulness of computed radiography is comparable to conventional films in the intensive care unit. The higher interobserver agreement rate for radiologic descriptors with computed radiography is an added advantage of the system. PMID- 9419459 TI - Noninvasive method for the detection of bowel ischemia: computed tomography detection of absorbed iodinated contrast material. AB - RATIONALE AND OBJECTIVES: Although systemic absorption of enterically administered iohexol and its excretion in urine has been previously documented in rats with ischemic bowel, a practical and sensitive method of detecting urinary iohexol has not been available. We proposed to detect the presence of iohexol in the urine of rats with normal and ischemic bowel by use of a computed tomography (CT) number increase in the bladder with the use of CT. METHODS: Anesthetized rats (250 g) underwent either sham laparotomy (n = 6), ligation of two vascular arcades to the proximal jejunum (n = 5), ligation of six vascular arcades to the proximal jejunum (n = 6), or ligation of the superior mesenteric artery (n = 6). Rats were hydrated with saline (3.2 ml/hr intravenously). Each received a 3-ml enteric bolus of isotonic iohexol. Serial CT scans and plain film radiographs of the bladder were performed at 2, 4, and 6 hr to detect systemic absorption of contrast from the gut. Urine iohexol concentrations were measured by capillary electrophoresis. CT number and iohexol concentration were compared with evidence from plain film radiographs of bladder opacification. Intestinal ischemia was graded histologically. RESULTS: Histologic evidence of ischemia was present in all six-arcade and five of six superior mesenteric artery (SMA)-ligated animals. No animals in the control or two-arcade group showed evidence of bowel ischemia. Statistically significant increases (P < 0.05) in bladder density were demonstrated in the six-arcade and SMA-ligated groups. No statistical difference was noted between the two-arcade ligation and control groups. CONCLUSIONS: Experimental intestinal ischemia was reliably detected by bladder opacification after administration of enteric contrast. CT detection of systemic absorption of enteric iohexol was more sensitive than plain film radiographs and may be useful in the diagnosis of intestinal ischemia, although it may not be specific for ischemia. PMID- 9419460 TI - Ultrasound monitoring of experimental ethanol injection into pig liver. AB - RATIONALE AND OBJECTIVES: In percutaneous ethanol injection therapy, uncontrolled spread of ethanol sometimes causes complications. This study tested the accuracy of ultrasonography in showing the spread of ethanol during injection in normal postmortem pig livers. METHODS: Methylene blue-stained 96% (vol) ethanol (0.5-2 ml) was injected into pig livers under sonographic control. The volumes of the sonographically determined areas infiltrated by the injections and the corresponding pale lesions seen after dissecting the livers were measured and compared. The locations of the stained areas were determined. RESULTS: Uncontrolled spread of ethanol was often seen sonographically as hyperechoic lines spreading centrifugally from the injection site. After dissection, nearly all vessels and ducts were stained blue, indicating a major escape of ethanol from the injection sites. The volumes of the sonographically determined injection areas were significantly larger (P < 0.001) than the lesions found after dissection. CONCLUSIONS: Ultrasonography may overestimate the spread of ethanol, making the survival of peripheral tumor cells possible. PMID- 9419461 TI - Searching for bone fractures: a comparison with pulmonary nodule search. AB - RATIONALE AND OBJECTIVES: We aimed to determine if the characteristics and principles of visual search described for the detection of pulmonary nodules apply to extremity fractures. METHODS: The eye positions of staff orthopedic radiologists, radiology residents, and medical students were monitored as they searched hand and wrist X-ray images for fractures and a chest image for nodules. RESULTS: More systematic scanning patterns were observed for experienced observers than inexperienced observers. Positive decisions for bone images were associated with prolonged gaze durations; prolonged gaze durations were significantly longer for false-negative versus true-negative decisions. Intercluster jump distances were found to be greater for chest images than bone images. CONCLUSIONS: A search for bone fractures can be qualitatively characterized by classifying observer scan paths, dwell times, and jump distances. Gaze duration can be a useful predictor of bone image locations containing potential missed fractures. Perceptual feedback could aid observers in the detection of inconspicuous fractures. PMID- 9419462 TI - Emergency department use of radiology services at a large urban teaching hospital. AB - RATIONALE AND OBJECTIVES: Little research has explored the use of radiologic services by emergency departments and the factors that influence use. This study aimed to identify and characterize these factors. METHODS: A total of 13,228 consecutive patient emergency charts from a large university emergency department were reviewed, and multiple parameters were entered into a database. The database was studied and statistical testing was done to identify significant parameters for patients who required imaging studies (X-ray group) and those who did not (non-X-ray group). RESULTS: Factors such as age, diagnosis, urgency of illness, and illness severe enough to require hospitalization were statistically significant in determining the need for a radiologic evaluation in the emergency setting. When these factors were equalized for the X-ray and non-X-ray groups by multivariate linear regression analysis, male sex was also found to be statistically significant. Factors such as race and presence or absence of health insurance were not statistically influential on multivariate analysis. CONCLUSIONS: Older age, diagnosis, and factors related to severity of illness affected the use of radiologic services in the emergency setting. Sex differences were also detected. PMID- 9419463 TI - Unobtrusively tracking eye gaze direction and pupil diameter of mammographers. AB - RATIONALE AND OBJECTIVES: The visual process that radiologists use for diagnosis is incompletely understood. This study developed techniques to unobtrusively track direction and pupil diameter of radiologists reading a wide variety of films. We evaluated the eye gaze patterns of mammographic experts to gain knowledge that might improve the rate of early detection of breast cancer. METHODS: A video camera with a near-infrared light filter is pointed at the mammographic expert who is reading mammograms. The video images are analyzed in real time on a personal computer to detect eye gaze direction and pupil diameter. Two separate trials were used: 1) to demonstrate the system's speed and ability to work with mammograms (a brief test with one mammographer was used) and 2) four mammographic experts evaluated 14 mammograms. RESULTS: In the first trial, the system successfully tracked the eye gaze of a mammographer who quickly recognized the patient case, with the pupil diameter briefly increasing 40%, and then the gaze direction dwelling in an area of microcalcifications. In the second trial, 66% of the false-positive results for films with masses were associated with long eye gaze dwells, whereas 33% of the prolonged dwells for films with microcalcifications were associated with true-positive diagnoses. CONCLUSIONS: This near-infrared light system successfully tracked the eye gaze direction and pupil diameter of mammographic experts evaluating films. The association of long eye gaze dwells with diagnostic accuracy varied with the type of object being viewed. In films with masses, false-positive diagnoses were associated with long dwells. In films with microcalcifications, true-positive diagnoses were associated with long dwells. PMID- 9419464 TI - Evaluation of the clinical performance of automated proton magnetic resonance spectroscopy in children. AB - RATIONALE AND OBJECTIVES: Because expeditious neuroimaging is imperative in pediatric patients, we evaluated automated procedures for proton magnetic resonance spectroscopy (1H MRS) of the brain of children. METHODS: 1H MRS was performed on a 1.5-T GE Signa. The protocol included stimulated echo-acquisition mode and spin-echo point resolved spectroscopy. The automated routine included adjustment of first-order gradient shims (x, y, z1) to optimize magnetic field homogeneity, transmit power, center frequency, receiver gain, and water suppression. All spectra were processed with the use of spectroscopy analysis software from General Electric on a Sun workstation. RESULTS: The use of the automated procedures reduced the length of our 1H MRS protocol by 50%. Magnetic field homogeneity was within our accepted standards (7 +/- 2 Hz). Water suppression was within range of our accepted factors (1000-10,000). However, on certain occasions, baseline distortions affected resonances in the 3.22-4.04 ppm range. CONCLUSIONS: Shortening of the time required for clinical 1H MRS will increase its application in evaluating children. PMID- 9419465 TI - A selected history of radiology apropos of the radiologic centennial. Part 5: The discovery of cesium 137: the untold story. PMID- 9419466 TI - Cost-effectiveness and safety of selective use of low-osmolality contrast media. AB - RATIONALE AND OBJECTIVES: We estimated our potential institutional savings by switching from universal to selective use of low-osmolality contrast media. METHODS: A total of 42,598 radiocontrast studies (26,595 with high-osmolality contrast agents and 16,003 with low-osmolality contrast agents [LOCA]) were performed over 42 months. Every radiocontrast reaction was classified and documented. All costs associated with the subsequent treatment of adverse reactions were recorded and added to the cost of contrast agents. The actual cost of contrast agent administration was determined and compared with the theoretical cost of total conversion to LOCA. RESULTS: The overall rate of adverse reactions was 1.18%. The cost of radiocontrast materials and treatment of adverse reactions was $2,599,593. Over the same period, total conversion to LOCA would have cost $5,968,507. CONCLUSIONS: Our institution saved an estimated $962,548 annually through the selective use of LOCA. The incremental cost of treating adverse reactions associated with selective use was minimal compared with the cost of total conversion to LOCA. PMID- 9419467 TI - Use of meta-analysis in the evaluation of imaging systems. PMID- 9419468 TI - Role of radiology in medical education: perspective of nonradiologists. AB - RATIONALE AND OBJECTIVES: We asked our nonradiologist colleagues to evaluate and comment on the most desirable format for radiology education. METHODS: Questionnaires were distributed to 631 nonradiologist physicians affiliated with the University of Minnesota and representing all medical specialties and academic ranks. Three hundred twenty-seven surveys were returned after one mailing. RESULTS: Residency was retrospectively noted to be indispensable for consolidating knowledge of radiology. The overwhelming majority of clinicians in all specialties believed that formal radiologic instruction should be mandatory for medical students (279 of 322; 87%). Film interpretation was believed to be an indispensable part of a medical student radiology rotation (226 of 321; 70%), but many clinicians indicated a need for additional training. A marked disparity in the perceived level of confidence in interpreting radiologic tests during medical school and residency between those who had and those who had not received formal radiologic instruction during medical school was evident. This difference in perceived level of confidence was present even among the most experienced clinicians. CONCLUSIONS: Collectively, the nonradiologist clinicians emphasized the need for a mandatory and clinically oriented radiology curriculum during medical school. PMID- 9419469 TI - Multiple ischemic infarcts versus metastatic disease. PMID- 9419470 TI - The business of radiology. PMID- 9419471 TI - Improved visualization of stimulated nodules by adaptive enhancement of digital chest radiography. AB - RATIONALE AND OBJECTIVES: Although digital radiography of the chest has a wide dynamic range, difficulties still remain in visualizing the acquired images faithfully on gray-scale displays (cathode-ray tube [CRT] displays), which have a much lower level of luminance than film view boxes. We propose an adaptive enhancement algorithm for digital chest radiography that provides faithful visualization of the chest on the CRT. METHODS: We investigated the contrast sensitivity of a CRT monitor and developed an image processing algorithm that compresses the dynamic range and enhances image contrast selectively in the mediastinal area and that transforms the gray scale to visualize the image by use of the full effective dynamic range of the CRT. We performed a receiver-operating characteristic (ROC) study by using simulated nodules to evaluate the clinical value of the proposed algorithm. RESULTS: The processed images provided improved visualization of both mediastinal and lung regions. The area under the ROC curve for retrocardiac or subdiaphragmatic nodule detection increased significantly (from 0.69 to 0.79; P < 0.05). The area under the ROC curve for lung nodule detection also increased (from 0.64 to 0.75; P < 0.1), although not to the level of statistical significance. CONCLUSION: The proposed algorithm allows improved visualization of nodules on digital chest radiographs with the CRT display. PMID- 9419472 TI - Ultrasonographic imaging and Doppler analysis of renal changes in non-insulin dependent diabetes mellitus. AB - RATIONALE AND OBJECTIVES: We used ultrasonographic imaging and Doppler analysis to assess renal changes in patients with non-insulin-dependent diabetes mellitus (NIDDM) and normal renal function, as established by normal serum creatinine levels and the absence of macroalbuminuria. METHODS: Renal parenchymal echogenicity, renal volume, and resistive index (RI) were blindly evaluated for 85 NIDDM patients and 42 age-matched control subjects (C). Results were analyzed and correlated with the following clinical parameters: patient age, duration of diabetes, blood pressure, blood glucose and cholesterol levels, and the presence of microalbuminuria. RESULTS: Normal renal parenchymal echogenicity was seen in all but one NIDDM patient; however, in comparison with C, diabetic patients had significantly higher renal volume (mean +/- standard deviation, 314.01 +/- 72.74 vs 227.64 +/- 58.76) and RI (mean +/- standard deviation, 0.71 +/- 0.05 vs 0.64 +/- 0.02). An RI higher than 0.70 was found in 55 of 85 (65%) NIDDM patients; an increased RI was directly correlated with patient age, whereas an inverse correlation existed between an increased RI and renal volume. No statistically significant differences were observed for the duration of diabetes, arterial hypertension, blood levels of glucose and cholesterol, and the presence of microalbuminuria. CONCLUSIONS: NIDDM patients with normal renal function show a significant increase in renal volume and RI in comparison with C. Demonstration of these findings may aid in the detection of early renal involvement in NIDDM patients. However, further investigations are needed to understand fully the correlation of such changes with the pathology of diabetic nephropathy and to provide an interpretation of the pathophysiologic mechanisms underlying changes in intrarenal vascular impedance in NIDDM patients. PMID- 9419473 TI - Demonstration of differences in vascular permeability in experimental tumors by use of 19F magnetic resonance imaging. AB - RATIONALE AND OBJECTIVES: In vivo assessment of tumor vascular permeability may provide useful information for chemotherapy treatment planning or for the assessment of treatment effectiveness. We aimed to assess vascular permeability in two tumor sublines as well as changes in vascular permeability with tumor growth by using 19F magnetic resonance imaging. METHODS: An emulsion of perfluorotributylamine was used as a tumor extravascular contrast agent for 19F MRI. The amount of emulsion that leaked into tumor interstitial space was analyzed qualitatively with imaging. A quantitative study of vascular permeability was done with a separate group of tumors by use of Evans blue dye. RESULTS: One tumor type was more permeable to both perfluorotributylamine emulsion and Evans blue than was the second tumor type. The difference was attributed to a difference in surface area for exchange. In larger tumors of both types, pooling of large amounts of perfluorocarbon occurred and was assumed to be attributable to hemorrhage or blood flow stasis or both. CONCLUSION: 19F MRI is capable of demonstrating the permeability of tumor vessels to macromolecular substances. PMID- 9419474 TI - Measurement of the ultrasonic attenuation of fat at high frequency. AB - RATIONALE AND OBJECTIVES: High-frequency ultrasound devices are often limited by a decreased depth of acoustic imaging caused by the increased attenuation of tissue at high frequencies. We investigated the role of adipose tissue in this phenomenon. METHODS: A substitution technique was used to calculate the ultrasonic attenuation (decibels per centimeter) of fresh samples of sheep rumen, omental fat, and back fat and swine back fat and various concentrations of bovine milk fat at 22 degrees C and 37 degrees C for frequencies of 15 and 20 MHz. RESULTS: The attenuation was significantly higher for sheep adipose tissue than for the intestinal wall, in descending order, omental fat, back fat, and rumen wall (P < 0.01). A correlation was found between bovine milk fat concentrations and attenuation at both frequencies (R2 > 0.9). The attenuation of adipose tissues decreased significantly with an increase in temperature (P < 0.01), whereas the attenuation of sheep rumen showed no significant change (P > 0.1). CONCLUSIONS: The ultrasonic attenuation of fat may contribute to limitations on the use of high-frequency ultrasound in clinical situations in which adipose tissue is present. PMID- 9419475 TI - Gallbladder wall thickening: an in vitro sonographic study with histologic correlation. AB - RATIONALE AND OBJECTIVES: We compared histologic sections with in vitro sonographic images for 40 resected gallbladder specimens to correlate the histopathologic processes with the sonographic appearance of the gallbladder wall. METHODS: In vitro sonographic images and histologic specimens were obtained by use of a specimen container with a micropositioner. An 8.5-MHz transducer and a scalpel were attached to the micropositioner to obtain the sonographic image and the histologic section. The sonographic images were analyzed for wall thickness and the number and echo texture of the visualized layers and then were correlated with the histologic sections. Each histologic specimen was interpreted as being normal or showing mild, chronic, acute, or gangrenous cholecystitis. RESULTS: One to four sonographic layers were observed in the gallbladder wall specimens. The number of wall layers was fairly evenly distributed among the different types of gallbladder wall inflammation. One to three sonographic layers were observed for most of the different types. In nearly all instances, the findings were attributable to either similar pathologic processes in two or more histologic layers or different pathologic processes in a single histologic layer. The gallbladder wall measured less than or equal to 3 mm in 89% of gallbladders with normal or mild inflammation, greater than 3-6 mm in 71% of cases of chronic cholecystitis, greater than 3 mm in 83% of cases of acute cholecystitis, and greater than 6 mm in 50% of cases of gangrenous cholecystitis. CONCLUSIONS: The sonographic layers in the inflamed gallbladder wall are determined by the pathologic changes present rather than by the normal histologic boundaries. However, because of the overlap of pathologic changes, we cannot predict the type of gallbladder wall pathology on the basis of the sonographic appearance. We found a trend toward gallbladder wall thickening for severely inflamed gallbladder walls. PMID- 9419476 TI - Urine profiles and kidney histology after intravenous injection of ionic and nonionic radiologic and magnetic resonance contrast media in normal rats. AB - RATIONALE AND OBJECTIVES: Previous studies showed that both high-osmolality and low-osmolality iodinated contrast media cause temporary albuminuria and enzymuria (presence of enzymes in urine) in normal rats. Whether the same is true with ionic high-osmolality and nonionic low-osmolality magnetic resonance (MR) contrast media is unknown. We studied urine profiles and histology after intravenous injection of four types of contrast media in rats with normal kidneys. METHODS: Urine profiles were monitored 4, 24, 48, and 72 hr after intravenous injection of saline, diatrizoate, iohexol, gadopentetate dimeglumine, and gadodiamide (4.59 mmol/kg of body weight) in normal rats. Each group included 20 male rats. After sacrifice, both kidneys were removed for examination by light microscopy (LM) and electron microscopy (EM). RESULTS: All four contrast agents caused a temporary (< 22 hr) increase in the excretion of albumin (2-5 times) and of cytoplasmic (30-100 times) and brush border (10-100 times) renal enzymes when compared with saline. The degree of albuminuria correlated well (r = 0.90) with the osmolality of the injected media, whereas the increased level of enzymuria was unrelated to the osmolality. No major differences in the enzymuric effects of the four agents were noted. LM revealed vacuoles in all kidneys exposed to radiologic contrast media but not in kidneys exposed to MR contrast media or saline. Slight vacuolation was revealed by EM after the use of MR contrast media, and significant vacuolation was evident via EM after the use of radiologic contrast media. No difference between ionic and nonionic media within each drug group was detected by either LM or EM. CONCLUSIONS: Transient renal effects are induced by both ionic and nonionic high-osmolality and low-osmolality radiologic and MR contrast media in normal rats. Both osmotic (e.g., albuminuria) and chemotoxic (e.g., enzymuria) mechanisms seem to be involved. From a morphologic point of view, the chemotoxic mechanisms seem to be of major importance. PMID- 9419477 TI - Contrast media-induced ventricular fibrillation: an experimental study of the effects of dimeric contrast media during wedged catheter injection in dogs. AB - RATIONALE AND OBJECTIVES: We investigated the cardiac effects of an ionic dimer, ioxaglate and two nonionic dimers, iotrolan, and iodixanol. METHODS: During a simulated wedged catheter situation, 22 ml of each contrast medium was injected into the left anterior descending branch of the left coronary artery in seven open-chested, anesthetized dogs. RESULTS: Of 13 injections with each contrast medium, ioxaglate induced ventricular fibrillation in 11 after 34 +/- 5 sec, iotrolan in 6 after 42 +/- 4 sec, and iodixanol in 3 after 61 +/- 1 sec. Ioxaglate markedly lengthened monophasic action potential duration in contrast medium-perfused myocardium. Iotrolan, and iodixanol induced biphasic changes, first lengthening and then shortening action potential duration. The electrophysiological changes occurred later when using iodixanol. CONCLUSIONS: The risk of ventricular fibrillation during long-lasting contrast media exposure to the myocardium, as in a wedged catheter situation, appears to be much lower with iodixanol compared with ioxaglate and also lower than when using iotrolan. PMID- 9419478 TI - Effects of radiologic contrast media on human endothelial and kidney cell lines: intracellular pH and cytotoxicity. AB - RATIONALE AND OBJECTIVES: Radiologic contrast media (CM) are hyperosmotic compounds injected undiluted into a patient's blood, in which they contact endothelial cells. For some types of cultured cells, the application of a hyperosmotic stimulus may cause intracellular pH (pHi) acidification that is related to the extent of hyperosmolality and that ultimately influences cellular function. Accordingly, endothelial and kidney cells, two types of cells known to be exposed to CM effects, were treated at relevant iodine concentrations with various CM (320-1500 mOsm) to determine whether cell exposure to CM can disturb the pH(i) and to examine the contribution of CM to cellular cytotoxicity. METHODS: Ionic (n = 3) and nonionic (n = 3) CM were compared. Changes in the pH(i) of human vascular endothelial and kidney cell lines were monitored by use of a pH-sensitive fluorescent dye (2',7'-bis(carboxyethyl)-5(6) carboxyfluorescein acetoxymethyl ester). The viability of cells treated with CM was determined by measuring the reduction of a tetrazolium salt (3-[4,5 dimethylthiazol-2-yl]-2,5-diphenlytetrazolium bromide) to violet formazan, a reaction that requires the activity of mitochondrial dehydrogenase; this measurement was made with a microplate reader. RESULTS: The pH(i) of endothelial and kidney cells exposed to 40-60 mg of iodine per milliliter of CM showed acidification (approximately 0.2 pH unit). Within minutes, gradual pH(i) alkalinization to baseline values occurred. The return to baseline values was slower with ionic compounds than with nonionic CM (P < 0.001). Nonionic agents caused less cellular damage than did ionic CM. CONCLUSIONS: The pH(i) is involved in the immediate intracellular transduction of CM effects in vitro. The exposure of cells to ionic CM is more detrimental than is exposure to nonionic CM, as demonstrated by disturbances in the cytosolic pH and by long-term effects on cell viability. PMID- 9419479 TI - Perfluoroctyl bromide as a blood pool contrast agent for computed tomographic angiography. AB - RATIONALE AND OBJECTIVES: We determined whether perfluoroctyl bromide (perflubron) could be used as a computed tomography (CT) angiographic agent by studying vessel visibility (celiac artery, superior mesenteric artery [SMA], and renal arteries) with spiral CT and three-dimensional (3D) reconstructions. METHODS: Five rhesus monkeys were examined with a perflubron emulsion (90% [w/v] perfluorochemical; administered intravenously at a dose of 1.5 ml/kg and at a rate of 0.5 ml/sec. Spiral CT was performed immediately and at 5 hr after injection. Three dimensional images of the aorta at the level of the celiac artery, SMA, and renal arteries were reconstructed and blindly rated 0-4 (0 = not seen; 4 = excellent visualization) by two observers. RESULTS: All the vessels had the best ratings immediately after injection: celiac artery, 2.8 +/- 0.42; SMA, 2.7 +/- 0.48; left renal artery, 2.1 +/- 0.99; and right renal artery, 1.2 +/- 1.03. The ratings after the 5-hr delay were as follows: celiac artery, 1.3 +/- 1.34; SMA, 1.5 +/- 1.08; left renal artery, 1.5 +/- 0.97; and right renal artery, 1.2 +/- 0.79. CONCLUSIONS: Spiral CT angiography with a perflubron emulsion successfully demonstrated all vessels immediately and at 5 hr after contrast agent infusion. Further refinements of the dose, rate, and reconstruction technique are expected to increase vessel visibility over this wide imaging window. PMID- 9419480 TI - Results of the 1993 survey of the American Association of Academic Chief Residents in Radiology. AB - RATIONALE AND OBJECTIVES: A survey of chief residents of academic radiology programs is conducted annually on behalf of the American Association of Academic Chief Residents in Radiology (A3CR2). Data are obtained to improve the training of diagnostic radiology residents and to increase the understanding of radiologists and their associates about issues of interest to radiologists in training. METHODS: Questionnaires were mailed to 133 accredited programs in the United States and Canada. A wide variety of demographic and common interest questions were asked. The analysis took into account geographic location of the responders and the size of the residency program. Comparisons were made to the data from prior years. RESULTS: Completed surveys from 93 programs (70%) were returned. The percentage of women residents is increasing. Important regional and size variations exist in several areas including salary, workload, prior clinical training, resident/fellow ratios, post residency plans, and call schedules. Although many chief residents feel knowledgeable about the health care system, opinions about the future of radiology and medical care are tentative. CONCLUSIONS: This survey provides important demographic information about academic radiology residency programs. The summary information regarding plans for fellowship training, resident call schedules, and opinions about socioeconomic issues may be useful for chief residents, program directors, and departmental chairmen. PMID- 9419481 TI - Residency and fellowship training in radiology: survey of all United States programs. AB - RATIONALE AND OBJECTIVES: We surveyed radiology training programs to determine current requirements for a clinical internship year, recent changes in the clinical internship requirement, current number of residents, percentage of residents with preradiology clinical experience, number of current first-year fellows, and percentage of residents entering fellowships. METHODS: Survey forms were sent to all 208 U.S. diagnostic radiology programs and were followed up by telephone and facsimile transmission. RESULTS: Survey response rate was 100%. One hundred programs (48%) require an internship, whereas 108 programs (52%) do not. Of programs without a clinical internship requirement, 33 (31%) have eliminated this requirement within the last 5 years. The clinical year requirement varied greatly by region. A total of 3983 residents are training at present, and 29% did not complete a clinical internship. Currently, 650 fellows are in training. Approximately 68% of graduating residents are entering fellowships. CONCLUSIONS: Many residency programs have recently discontinued the clinical internship year requirement. Changes in Medicare reimbursement of resident salaries may be one factor promoting this change. During the past 4 years, the number of radiology residents training per year has increased. The percentage of residents entering radiology without an internship year has increased by 6% during the past 4 years. An increasing number of graduating residents are entering fellowships. PMID- 9419482 TI - Improving the efficiency and service of computed tomographic scanning. AB - RATIONALE AND OBJECTIVES: This study quantified the impact on service and costs of operational changes in CT. METHODS: Operational and financial analyses were performed in 1988 and 1991 after process flow and management changes were made. RESULTS: The backlog decreased from about 6 weeks to 1 week. The cost per scan decreased 25%. Volume increased 50%. Two technologists were assigned per scanner doing some steps of the process in parallel rather than sequentially. Decreasing throughput time accounted for three fifths of the cost decrease. The reduction in number of scanners and hours of operation, change in radiologists' practice patterns, coordination of scheduling, CT priority for escort personnel, and personnel changes accounted for two fifths of the cost decrease. CONCLUSIONS: It is possible to simultaneously decrease costs and improve the quality of service by careful operations analysis and management. Operational changes had synergistic effects that allowed more improvement than might be anticipated. Advisable management changes may be counterintuitive. PMID- 9419484 TI - Termination of a mandatory radiology clerkship at the Albert Einstein College of Medicine: a survey of student opinion. AB - RATIONALE AND OBJECTIVES: The Albert Einstein College of Medicine has replaced a mandatory third-year radiology clerkship with a new family medicine rotation. This was done, ostensibly, to influence student career choice. We surveyed students to quantify their perceptions about the former radiology course. METHODS: Questionnaires were sent to 498 present and former Albert Einstein College of Medicine students. Eight questions attempted to quantify the students' perceptions about the educational value of the radiology clerkship and assess its impact on subsequent career choice. RESULTS: The mandatory clerkship taught the fundamentals of X-ray interpretation and the most efficacious ways of imaging disease processes. Students viewed the clerkship as useful and important for a well-rounded medical education. Most would have taken the course as an elective had it only been offered as such and would now encourage junior students to do likewise. The mandatory radiology clerkship did not influence the career choice of most survey respondents. CONCLUSIONS: The former mandatory radiology clerkship had been an important part of student education at Albert Einstein College of Medicine, and its termination is unlikely to affect student career choice. PMID- 9419483 TI - International Workshop on Standardization in Clinical Magnetic Resonance Spectroscopy Measurements: proceedings and recommendations. PMID- 9419485 TI - Point-counterpoint. Plagiarism: what is it, whom does it offend, and how does one deal with it? PMID- 9419486 TI - Worsening right flank pain over a 24-hr period. AB - AML is a benign renal tumor composed of variable quantities of mature vascular, smooth muscle and fatty elements. They occur as an isolated finding, classically in middle-aged females, or in association with tuberous sclerosis. When symptomatic, they typically present with flank pain secondary to hemorrhage. CT is the diagnostic imaging modality of choice. The diagnosis can usually be made based on the recognition of fat within the lesion. When discovered, asymptomatic lesions are generally monitored by follow-up imaging studies, and if they remain stable, no intervention is required. Arterial embolization has become the recommended treatment of choice in some instances, particularly in cases with associated hemorrhage. PMID- 9419487 TI - Career planning and development. PMID- 9419488 TI - 1H magnetic resonance spectroscopy characterization of neuronal dysfunction in drug-naive, chronic schizophrenia. AB - RATIONALE AND OBJECTIVES: We investigated the proton metabolism of right prefrontal white matter in drug-naive, chronic schizophrenic patients (n = 23), compared with healthy normal control subjects (n = 10), by using localized, water suppressed in vivo 1H magnetic resonance (MR) spectroscopy. METHODS: All 1H MR spectroscopy examinations were performed on a 1.5-T MR imaging/MR spectroscopy system by using a point-resolved spectroscopy pulse sequence for localized volumes of 2 x 2 x 2 cm3. Proton metabolite ratios relative to creatine (Cr) were obtained using a Marquart algorithm. RESULTS: Drug-naive, chronic schizophrenic patients demonstrated a decrease in the N-acetylaspartate (NAA):Cr and choline (Cho):Cr ratios and an increase of the complex of gamma-aminobutyric acid (GABA) and glutamate (Glu)-containing ratio [(GABA + Glu):Cr] as compared with normal control subjects. CONCLUSION: Results suggest that the reduction of NAA and Cho may indicate neuronal dysfunction and that the elevation of GABA and Glu may play a role in chronic schizophrenia. 1H MR spectroscopy may be a useful modality in research and in the clinical evaluation of chronic schizophrenic patients. PMID- 9419489 TI - Influence of clinical history on perception of abnormalities in pediatric radiographs. AB - RATIONALE AND OBJECTIVES: We tested whether having clinical information would improve perception or simply decision making. METHODS: Sixty-four pediatric chest and abdominal radiographs, half of which had abnormalities, were presented to nine radiologists under one of two conditions. In one condition, history consistent with abnormalities actually present for positive cases was provided for positive and matched negative cases before inspection. In a second condition, this information was provided only after inspection was completed and the radiograph was no longer available. Because detailed visual memory is short lived, the image information was no longer available when the history was provided after inspection. A control condition measured detection without history. RESULTS: Detection was significantly better with history provided before inspection. Detection did not differ for history provided after inspection and inspection without history. CONCLUSION: The only difference between conditions with history was in whether history influenced perception; history affected decision making in both conditions. Clinical history affected perception in interpreting radiographs, not simply decision making. PMID- 9419490 TI - Reproducibility of five anorectal morphologic measurements in defecography. AB - RATIONALE AND OBJECTIVES: We evaluated the inter- and intraobserver reproducibility of measuring five morphologic parameters of the anorectum in defecography (evacuation proctography). METHODS: Measurements from 42 defecographic studies were statistically analyzed. The parameters measured during resting, squeezing, and straining included two anorectal angles (posterior and axis), maximal width of the anal canal, maximal width of the rectal lumen, and size of the rectocele. RESULTS: The results demonstrated only fair interobserver agreement (kappa = 0.22-0.38) for almost all measurements of the five morphologic parameters. There were high correlations (kappa = 0.62-1.00) among most intraobserver measurements. CONCLUSION: For defecographic measurement, the five parameters we studied have relatively poor clinical value because of high inter- and intraobserver inconsistency. PMID- 9419491 TI - Transcatheter hepatocyte transplantation: preclinical studies of anatomic consequences in the portal vascular bed. AB - RATIONALE AND OBJECTIVES: Intrasplenic transplantation deposits hepatocytes in host hepatic sinusoids with amelioration of chronic liver failure and genetic deficiency states. Because portal resistance can be altered by intrasinusoidal transplanted cells, we examined whether hepatocyte recipients would develop deleterious portal hypertension or portosystemic collaterals. METHODS: Syngeneic hepatocytes in suspension were transplanted into recipient rats by transcatheter injection into the splenic parenchyma. Subjects included recipients of 2 x 10(7) hepatocytes representing approximately 3% of the host hepatic mass, recipients of 7.5 x 10(7) hepatocytes representing approximately 12.5% of the host hepatic mass, normal control rats, and positive control rats with portal hypertension induced by partial portal vein constriction. Portal pressures were recorded with a sensitive transducer, portosystemic collaterals were demonstrated with direct splenoportography, and survival of transplanted cells was determined with an endogenous dipeptidyl peptidase IV reporter gene. RESULTS: In normal rats, the portal pressure was 6.25 +/- 1.9 mm Hg with no portosystemic collaterals. By contrast, portal pressures were significantly increased in portal vein constricted rats, 20.7 +/- 3.9 mm Hg (P < 0.001), with extensive portosystemic collaterals. In hepatocyte recipients, portal hypertension observed during transcatheter cell injection but proved transient. When animals were examined up to 16 weeks after hepatocyte transplantation, portal pressures were in the normal range (after 2 x 10(7) cells, 7.5 x 2.6 mm Hg; after 7.5 x 10(7) cells, 9.5 +/- 4.2 mm Hg, P = not significant). No portosystemic collaterals developed in hepatocyte recipients at various times up to 8 months after transplantation. Transplanted hepatocytes expressing the reporter gene were present in recipients with assimilation in host hepatic cords. CONCLUSION: Despite injection of a massive number of cells, transcatheter hepatocyte transplantation was devoid of any significant portal vascular alterations or toxicity in recipients. These findings are consistent with assimilation of transplanted hepatocytes into host hepatic cords and will facilitate therapeutic applications in metabolic diseases or acute liver failure. PMID- 9419492 TI - Semiautomatic evaluation of left ventricular diastolic function with cine magnetic resonance imaging. AB - RATIONALE AND OBJECTIVES: Cine magnetic resonance (MR) imaging is a relatively new technique that can be used to study cardiac function with high spatial and temporal resolution. However, detailed functional analysis of the entire cardiac cycle with cine MR imaging is time consuming and labor intensive. We analyzed diastolic function using a semiautomatic routine that reduces the time necessary for analysis. METHODS: Twenty subjects (10 normal control subjects and 10 patients with isolated diastolic dysfunction) were examined. Short-axis cine MR images were obtained at 32 phases of the cardiac cycle. A semiautomatic boundary finding routine was used to determine left ventricular (LV) volumes at each phase. Volume-versus-time and first-derivative curves were created from these data. Several parameters derived from the MR imaging curves were used to characterize diastole. RESULTS: Two parameters--the ratio of early peak filling rate to late peak filling rate and the percentage of filling during early diastole--perfectly distinguished subjects with LV diastolic dysfunction from the normal control subjects. The semiautomatic analysis method substantially reduced the time necessary for analyzing the MR imaging data, compared with manual analysis. CONCLUSION: Cine MR imaging, especially with time-saving techniques such as our service automatic analysis method, has promise as a research and clinical tool in evaluating LV diastolic function. PMID- 9419493 TI - The relationship of age, gender, handedness, and sidedness to the size of the corpus callosum. AB - OBJECTIVES: We correlated the area and size of the corpus callosum, as measured by MR imaging, with the individual's handedness, sidedness, age, and gender. MATERIALS AND METHODS: A total of 117 patients (59 male, 58 female) aged 15-75 years were selected for this study. This included 12 persons who were either left handed or ambidextrous. Each patient was tested extensively to determine handedness and sidedness. Callosal areas and thickness were measured and correlated with brain size. RESULTS: The body of the corpus callosum decreases in size with age and is larger in right-handed persons. The cross-sectional areas of the genu, splenium, and corpus callosum, overall, do not vary significantly with respect to age, gender, sidedness, or handedness. CONCLUSIONS: The size of the corpus callosum consistently decreases with age. Otherwise, few statistical differences in callosal size relate to gender, sidedness, or handedness. PMID- 9419494 TI - Effects of radiographic contrast media on leukocyte phagocytosis. AB - RATIONALE AND OBJECTIVES: Differences in leukocyte phagocytosis following exposure to different classes of radiographic contrast media (RCM) may help in the development of less toxic alternatives and be useful as a guide to contrast selection in certain patient groups. The effect of RCM on leukocyte phagocytosis of Escherichia coli was examined. METHODS: Cell population phagocytosis and individual cell phagocytic activity were assessed in a control group and in samples exposed both to the ionic RCM diatrizoate and ioxaglate and to the nonionic RCM iohexol and iotrolan by using a flow cytometer. RESULTS: The percentage of granulocytes undergoing phagocytosis was 88.4% in the control population. Following exposure to RCM, this value fell to 79.2% with iohexol, 77.6% with iotrolan, 70.5% with diatrizoate, and 68.7% with ioxaglate. The number of bacteria phagocytosed by active leukocytes was not affected by RCM. CONCLUSION: RCM adversely affect the percentage of granulocytes involved in phagocytosis but not the number of pathogenic bacteria that are phagocytosed by individual granulocytes. PMID- 9419495 TI - Pancreatic contrast enhancement with Mn-bis pyridoxal ethylene diamine diacetic acid and the influence of a hormonal stimulation of the pancreas in pigs. AB - RATIONALE AND OBJECTIVES: Pancreatic and hepatic contrast enhancement after different modes of administration of Mn-bis pyridoxal ethylene diamine diacetic acid (Mn-DPDP) and the influence of the hormones secretin and cholecystokinin on the uptake of this new contrast agent were studied by using magnetic resonance (MR) imaging and atomic absorption spectroscopy (AAS) examinations carried out on 13 young domestic pigs. METHODS: Mn-DPDP was administered either as bolus injection or 30-min infusion. The course of contrast enhancement was noted. MR imaging was carried out with or without pancreatic stimulation. The Mn content in pancreatic and hepatic tissue was analyzed by using AAS. RESULTS: Mn-DPDP caused a significant increase (66 +/- 40.1%) in pancreatic signal intensity, an increase in hepatic signal intensity (78 +/- 23.0%), and a significantly higher Mn content on tissue samples of both organs. The results showed a wide individual range. With infusion of Mn-DPDP, pancreatic enhancement was slower, with a peak at 82 +/ 34 min compared with 54 +/- 51 min following bolus injection. The Mn content of both hepatic and pancreatic tissue determined by AAS seemed to be higher after infusion. An incomplete washout of Mn after 24 hr was demonstrated. Hormonal stimulation of the pancreas further increased pancreatic signal intensity but did not influence hepatic contrast enhancement. CONCLUSION: Infusion is the method of choice for administering Mn-DPDP because it causes an increase in hepatic signal intensity but does not change pancreatic enhancement. Although pancreatic hormonal stimulation intensifies enhancement, the increase is not statistically significant, and this practice will not contribute to day-to-day clinical practice. PMID- 9419496 TI - Role of sodium addition to nonionic contrast medium in preventing ventricular fibrillation during coronary arteriography in dogs. AB - RATIONALE AND OBJECTIVES: We studied the effect of the addition of sodium to nonionic contrast medium (CM) on the incidence of ventricular fibrillation (VF) during coronary arteriography in dogs. METHODS: We infused 20 ml (0.5 ml/sec) of iohexol, iohexol plus 30 mmol of Na+ per liter, and NaCl-Ringer's acetate in randomized order through a wedged catheter placed in the right coronary artery (RCA) or in the left anterior descending coronary artery (LAD) in 12 anesthetized dogs. In addition to electrocardiographic and hemodynamic measurements, epicardial monophasic action potential durations and ventricular activation times were recorded during infusions into the LAD. RESULTS: All infusions with iohexol into the RCA and the LAD (n = 16) caused VF. Seven of 19 infusions with iohexol plus 30 mmol of Na+ per liter caused VF. Infusions with iohexol plus 30 mmol of Na+ per liter that did not cause VF lengthened monophasic action potential durations and increased ventricular activation times more in the CM-perfused area than in the control area. CONCLUSION: The addition of sodium to iohexol reduces the incidence of VF when infused through a wedged catheter. The protective mechanisms may be attributable to a lengthened repolarization phase and an increased activation time in the CM-perfused area. PMID- 9419497 TI - Magnetic resonance assessment of microvascular patency in reimplanted rabbit ears. AB - RATIONALE AND OBJECTIVES: Magnetic resonance angiography (MRA) provides a means of noninvasive vascular imaging of microvascular vessels. Before conducting comparative studies and value assessments of a new method, it is necessary to evaluate the reproducibility and reliability of the technique. In an experimental study with five rabbits, we investigated the feasibility of MRA imaging of microsurgical vessels. We also attempted to demonstrate the reproducibility of the method. METHODS: We performed MRA imaging of the vascular tree of five New Zealand white rabbit ears, including normal ears, after microvascular reimplantation of the ear and in occlusion experiments on the ear. Scans of four reimplanted ears were performed after the microvascular procedure. In four occlusion experiments, the central vessels were totally occluded by a ligature, and scans were carried out prior to and immediately after occlusion. We used a fast imaging with steady-state precession (FISP) three-dimensional (3-D) rephased dephased sequence (28-msec repetition time [TR], 18-msec echo time [TE], and a 20 degrees flip angle). RESULTS: In normal rabbit ears and in the reimplantation cases, MRA imaging was able to show the flow within the vascular tree. In the reimplanted ears, MRA images confirmed patency in all anastomosed vessels. The diagnosis of occlusion appeared to be secure in the fully completed experiments (two of four) when it was based on the absence of a signal in the occluded vessels. Because of poor image quality during preligation scanning, we prematurely canceled two experiments. CONCLUSIONS: This study demonstrates that MRA imaging could correctly identify 1-mm vascular channels. Because of the long acquisition time, motion could compromise the reliability of the technique in human subjects. PMID- 9419498 TI - Survey of overhead assessments of academic health center radiology departments. PMID- 9419499 TI - An index for diagnostic accuracy in the multiple disease setting. AB - RATIONALE AND OBJECTIVES: Evaluation of diagnostic accuracy in the clinical environment should entail some assessment of performance in patients with multiple abnormalities. Although receiver operating characteristic (ROC) curves often are used to assess the diagnostic accuracy of imaging systems, the concept is not easily generalizable to patients with multiple abnormalities. I propose a measure of diagnostic accuracy that is a generalization of the area under the ROC curve for a single disease. METHODS: The proposed measure of diagnostic accuracy is a weighted average of the area under individual ROC curves for the single disease setting and of components representing areas under ROC curves constructed for patients with multiple diseases. Several options are discussed for scoring the presence of abnormality for patients who have two or more abnormalities. RESULTS: Methods of estimating diagnostic accuracy are demonstrated on a set of data in which more than one third of the abnormal cases included multiple abnormalities of chest disease. CONCLUSION: An easy-to-use method is given to estimate diagnostic accuracy in the multiple abnormality setting. This should make it easier to incorporate cases with multiple abnormalities when assessing the diagnostic accuracy of imaging systems. PMID- 9419500 TI - Longitudinal comparison of multimedia textbook instruction with a lecture in radiology education. AB - RATIONALE AND OBJECTIVES: The purpose of this prospective randomized study was to compare the long-term instructional effectiveness of a computer-based radiology multimedia textbook (MMTB) with that of a traditional lecture. METHODS: Volunteer faculty/fellows and residents were randomly assigned to either a computer-based MMTB group or to a lecture group. The course content for each instructional group was the same. Pretests, posttests, and 1-year long-term retention tests were administered to both groups. The same 10 questions were on all tests. The resulting data were analyzed using analysis of variance procedures available on the Statistical Analysis System. RESULTS: A comparison of the long-term instructional effectiveness of an MMTB versus a lecture showed that the MMTB computer instructional method was at least comparable in spite of the initial short-term appearance of lecture superiority. CONCLUSION: These results suggest a promising future for MMTB and other forms of computer-based education in radiologic instruction for medical students and radiologists. PMID- 9419501 TI - The critical roles of research and education in health care system reform. PMID- 9419502 TI - Universal use of low-osmolar contrast media: a European perspective. PMID- 9419503 TI - Pathologic fracture in a 66-year-old man. PMID- 9419504 TI - Point-counterpoint. Lymphography for staging lymphomas: is it still a useful procedure? PMID- 9419505 TI - The "weakening" job market. PMID- 9419506 TI - AUR Memorial Award. Identification of myocardial cell death in reperfused myocardial injury using dual mechanisms of contrast-enhanced magnetic resonance imaging. AB - RATIONALE AND OBJECTIVES: Because the magnitude of dysprosium-induced signal loss depends on the microheterogeneity of its distribution (exclusion from intracellular space), we proposed that loss of myocardial cell integrity would be reflected by decreased potency of dysprosium in the injured compared with normal myocardium. We measured the effect of dysprosium on magnetic resonance (MR) imaging signal intensity of reperfused infarcted and nonischemic myocardium and related it to tissue concentration of the contrast media. METHODS: Rats were subjected to 1 hr coronary artery occlusion followed by 1 hr reperfusion. After 45 min of reflow, group 1 (n = 9) received 1.0 and 0.2 mmol/kg dysprosium diethylenetriamine pentaacetic acid-bismethylamide (Dy-DTPA-BMA) and gadodiamide (Gd-DTPA-BMA), respectively. Group 2 (n = 7) received no contrast agents. Excised hearts were imaged with spin-echo T1- and T2-weighted sequences. After imaging, hearts were stained (triphenyltetrazolium chloride) to define the injured zones. Concentrations of Dy-DTPA-BMA and Gd-DPTA-BMA in regional myocardial tissue were determined by induction coupled plasma-atomic emission spectrometry. Separate groups received one or the other contrast medium alone to control for potential error from the mixed effects of the two agents. RESULTS: Gd-DTPA-BMA delineated reperfused infarcted myocardium as a bright zone on T1-weighted images, thus indicating delivery of the agent and reperfusion at the tissue level. Dy-DTPA-BMA delineated the reperfused infarction as a bright region by decreasing the signal intensity of nonischemic myocardium significantly more than that of injured myocardium, despite being present in greater concentration (by 2.46-fold) in the injured myocardium. CONCLUSION: These findings are consistent with the hypothesis that the failure of myocardial cells to exclude the dysprosium compound is responsible for the diminished potency of dysprosium to cause MR imaging signal intensity loss in reperfused myocardial infarction. The combination of the two contrast media may define reperfusion of the myocardium at the tissue level (Gadolinium distribution) and the presence and extent of myocardial necrosis (diminished dysprosium effect) in reperfused myocardial infarctions. PMID- 9419507 TI - Joseph E Whitley, MD, Award. Computer-assisted instruction with interactive videodisc versus textbook for teaching radiology. AB - RATIONALE AND OBJECTIVES: We compared computer-assisted instruction with interactive videodisc (CAI-videodisc) with a textbook to study differences in educational efficacy, time spent, and subjective preferences. METHODS: Two modules of CAI-videodisc, one concerning the radiology of arthritis and the other skeletal trauma, were prepared specifically for advanced medical students and junior house staff. Because the modules were derived from an actual textbook, we were able to conduct a controlled comparison. Our participants were 103 third- and fourth-year medical students taking a required 2-week clerkship in diagnostic radiology. They were assigned as part of their coursework the CAI-video-disc version of one module and the textbook version of the other. Pre- and posttests were administered. RESULTS: The mean scores improved from pre- to posttest after students used both modules in either version (P < 0.001). The gain in score was greater for students using CAI-videodisc; this difference was small only for the arthritis module (P < 0.092), but it was large for the trauma module, even when adjusted for differences in time spent learning (P < 0.001). Significantly more time was spent on the CAI-videodisc versions than the textbook versions (P < 0.004). Subjectively, 43% of the students preferred the CAI-videodisc, 45% preferred the textbook, and 11% had no preference. CONCLUSION: Medical students learned more radiology using a CAI-videodisc program than reading a textbook, but spent more time doing so. Their subjective preferences were equally split. PMID- 9419508 TI - Observer detection performance in radiology using a retransmission-free network communication protocol. AB - RATIONALE AND OBJECTIVES: We measured the effect of image data loss on diagnostic accuracy to test the possibility of using a retransmission-free network communication protocol for transferring radiologic images. METHODS: Mammograms transferred over a simulated network with 0%, 15%, and 25% transmission packet loss were presented randomly to 10 observers who typically read mammograms. Observers reported on the presence or absence of microcalcification clusters and the number of calcifications per cluster. RESULTS: Performance with 15% loss did not differ from performance with 0% loss. The 25% loss condition resulted in a significant decrease in performance. Accuracy of counting individual microcalcifications was high in all three loss conditions. CONCLUSION: Up to 15% packet loss can be tolerated without affecting diagnostic performance. These data suggest that in some radiologic applications retransmission-free communication protocols may be useful. PMID- 9419509 TI - Pharyngeal solid-state manometry catheter movement during swallowing in dysphagic and nondysphagic participants. AB - RATIONALE AND OBJECTIVES: The movements of the soft palate and the larynx are crucial in pharyngeal manometry because of the potential risk of manometry sensor dislocation. METHODS: Twenty dysphagic patients and 20 nondysphagic volunteers were examined with simultaneous videoradiography and intraluminal pharyngeal solid-state manometry. The movements of the manometric sensors were analyzed from lateral videorecording. RESULTS: Two different types of catheter movement were found. The sensor in the upper esophageal sphincter (UES) could either be lifted cranially during the closure of the soft palate (type 1) or stay unaltered in the sphincter until the beginning of the laryngeal elevation and then follow the sphincter cranially during laryngeal elevation with no previous response to soft palate closure (type 2). Type 1 movement was found in eight of 20 patients but in only one of the 20 volunteers. The resting pressure of the upper esophageal sphincter was significantly higher in type 2 (P = 0.004). Nineteen of the 20 patients with a high resting pressure of the UES (83+ mm Hg) were found to have the type 2 movement. CONCLUSION: High resting pressure in the UES permitted the sphincter to grasp the manometry catheter and caused the sensor to follow the cranial movement during laryngeal elevation. Sensor movement is important during pharyngeal manometry, and sensor dislocation out of the sphincter can be misinterpreted as sphincter relaxation. Simultaneous videoradiography provides control of sensor positioning and allows for correction. PMID- 9419510 TI - An ellipsoidal shell model for volume estimation of the right ventricle from magnetic resonance images. AB - RATIONALE AND OBJECTIVES: I developed a volume estimation technique for the crescentic volume of the right ventricle (RV) of the heart. A geometric model was desired to avoid the lengthy data collection and reduction required by Simpson's rule. METHODS: An ellipsoidal shell model was developed that requires only simple mathematics and that resembles the RV shape. RV cast volumes were obtained by water displacement and Simpson's rule and model-based calculations using magnetic resonance (MR) imaging. RESULTS: Model-based estimates correlated well with water displacement volumes (r = 0.924), with a slope not significantly different from unity. Simpson's rule results showed a higher correlation (r = 0.994), but it required longer acquisition and processing. Geometric irregularity in the RV shape required no modification in mathematics. CONCLUSION: The model provides reliable RV volume estimates from two MR image planes. The mathematics provides a simple approach to a relatively complex, crescentic shape. Short times for data acquisition and analysis suggest the potential for time savings during routine clinical measurements. PMID- 9419511 TI - Percutaneous computed tomography lymphography in the rabbit by subcutaneously injected nanoparticulates. AB - RATIONALE AND OBJECTIVES: Surgical lymphangiography is infrequently used in staging cancer because of its inherent limitations. Radiopaque nanoparticulates target lymph nodes draining interstitial tissues and could make percutaneous lymphography feasible. METHODS: Experimental nanoparticulate contrast agent formulations were injected subcutaneously in the forepaw or hindpaw of normal rabbits or rabbits with induced reactive nodal hyperplasia. Axillary and popliteal nodes were imaged with thin-section computed tomography (CT) using quantitative methods to measure node enhancement. Dose-response (0.1-2.0 ml) and time course (4 hr to 10 weeks) of enhancement were assessed. RESULTS: Nodal enhancement above 100 Hounsfield units was consistently obtained. Enhancement was significantly related to dose and peaked at 10 hr with slow washout over the observation period. Nodes with reactive hyperplasia were larger and had heterogeneous enhancement patterns distinctly different from normal nodes. CONCLUSION: Percutaneous CT lymphography effectively depicts the macroscopic intranodal architecture in rabbits. PMID- 9419512 TI - Time-lapse quantitative computed tomography lymphography: assessing lymphatic function in vivo. AB - RATIONALE AND OBJECTIVES: Immobility and massage produce different local limb lymph flow rates. We studied their influence on accumulation of radiopaque nanoparticulates in regional lymph nodes of normal rabbits. METHODS: Quantitative lymphography at 10-min intervals was used to follow the transport of subcutaneous (s.c.) nanoparticulates produced from insoluble esters of diatrizoic acid. In one design, both hindpaws received 0.5 ml of nanoparticulate s.c., and one hindpaw was massaged. In a second design, one hindpaw was injected and massaged while imaging the popliteal, presacral, and paraaortic nodes every 10 min. RESULTS: Gentle massage rapidly increased popliteal node accumulation in comparison with the immobile limb. On the massaged side, mean Hounsfield (HU) units, maximum Hounsfield units, and calculated iodine were significantly greater at 10 min and all subsequent times. In the node transfer experiments, it took 12, 30, and 45 min, respectively, to obtain 100-HU mean attenuation; 200-HU maximum attenuation thresholds were achieved at 20, 47, and 69 min, respectively. CONCLUSION: Quantitative computed tomography lymphography reflects local lymph physiology. Gentle massage of the s.c. injection site is a powerful lymphotropic stimulus. PMID- 9419513 TI - Hydrostatic versus oleic acid-induced pulmonary edema: high-resolution computed tomography findings in the pig lung. AB - RATIONALE AND OBJECTIVES: We evaluated the differences between combined hydrostatic and hypervolemic edema and oleic acid-induced edema on high resolution computed tomography (HRCT) scans. METHODS: Twelve anesthetized and ventilated pigs were studied. Hydrostatic edema was induced by ligation of the abdominal aorta and infusion of normal saline (n = 4); permeability edema was induced by intravenous injection of oleic acid (n = 4). Four pigs were studied as normal controls. Serial scans were obtained before and after induction of edema at a constant position in the caudal lobe of the lung. The distribution of edema was assessed visually. Cross-sectional areas (CSAs) of the pulmonary artery and vein were measured both at the lobar and segmental levels. RESULTS: Gravity dependent opacity, peribronchovascular fluid collection, prominent centrilobular core, thickening of the interlobular septa, and air-space consolidation at the dependent site were the sequential HRCT findings of hydrostatic edema. Randomly distributed, diffuse patchy high attenuation areas with a tendency for predilection in the subpleural and peripheral areas of the secondary lobule were the findings of oleic acid-induced edema. Hydrostatic edema increased the mean CSAs of the lobular vein by 137.8% +/- 78.7, but oleic acid edema decreased the mean CSAs by 33.2% +/- 22.7. Changes in the mean CSAs of the pulmonary arteries were not significant. The mean vein-to-artery ratio increased significantly in hydrostatic edema but decreased in oleic acid edema. CONCLUSION: HRCT findings for hydrostatic and oleic acid-induced pulmonary edema differed both in distribution of edema and in pulmonary vascular response. PMID- 9419514 TI - Nanoparticulate computed tomography contrast agents for blood pool and liver spleen imaging. AB - RATIONALE AND OBJECTIVES: We investigated the properties of a group of iodine containing, insoluble compounds formulated as nanoparticles for use as potential blood pool and liver-spleen contrast agents. METHODS: High-resolution, quantitative computed tomography (CT) was performed prior to and at intervals following the intravenous administration of the contrast agents to rabbits. Time density characteristics for three organs were evaluated. RESULTS: Excellent enhancement of blood (< or = 232 Hounsfield units [HU]), liver (< or = 263 HU), and spleen (< or = 350 HU) was achieved at the administered dose of 3.0 ml/kg. The composition of the agents influenced the biodistribution, as well as the residence time in blood, and time to peak enhancement in liver. CONCLUSION: Iodinated nanoparticulate compounds are promising CT contrast agents. Development of agents with desirable pharmacokinetic and biodistribution profiles may permit application-specific contrast enhancement. PMID- 9419515 TI - Iodinated nanoparticles for indirect computed tomography lymphography of the craniocervical and thoracic lymph nodes in normal dogs. AB - RATIONALE AND OBJECTIVES: We evaluated the imaging characteristics of an interstitially or intraperitoneally delivered iodinated particulate contrast agent for computed tomography (CT) lymphography of the craniocervical and thoracic lymph nodes. METHODS: We injected 2-4 ml of 15% wt/vol iodinated nanoparticle suspension subcutaneously, submucosally, or intraperitoneally in eight normal dogs. CT and plain radiographic images were obtained prior to contrast administration and 4 hr, 24 hr, and 7 days after injection. Correlation was made to detailed postmortem assessment. RESULTS: CT images showed enhancement of regional nodes draining injection sites. Mean attenuation of opacified nodes was 313 +/- 297 (mean +/- standard deviation), 536 +/- 453, and 492 +/- 372 Hounsfield units at 4 hr, 24 hr, and 7 days postinjection, respectively. Lymph node opacification on CT images correlated well with node location found at postmortem. CONCLUSION: Craniocervical and thoracic lymph nodes can be effectively opacified from interstitial or intraperitoneal delivery of this iodinated nanoparticulate contrast agent. PMID- 9419516 TI - Evaluation of a breast biopsy phantom for learning freehand ultrasound-guided biopsy of the liver. AB - RATIONALE AND OBJECTIVES: We evaluated whether a breast biopsy phantom device would aid in the development of skills in freehand ultrasound liver biopsy. METHODS: Three radiologists who were inexperienced in freehand biopsy of the liver were observed. Each radiologist was timed and scored during attempts to biopsy lesions created in a beef liver. The time required for biopsy and the success of each pass was recorded. A commercially available breast biopsy phantom was then used by each of these radiologists during two 20-min practice sessions. Posttraining testing on the beef liver was performed in the same manner as pretraining testing. RESULTS: Freehand biopsy practice using the breast biopsy phantom resulted in a reduction in the mean time required for biopsy from 32 to 17 sec. Each of the three subjects reduced the mean time required for successful biopsy after training using the breast biopsy phantom. The total number of lesions missed was reduced from 14 of 43 to 0 of 45. CONCLUSION: Practice using the ultrasound breast biopsy phantom improves performance in freehand ultrasound biopsy of the liver. PMID- 9419517 TI - Automated segmentation of digitized mammograms. AB - RATIONALE AND OBJECTIVES: Fast and reliable segmentation of digital mammograms into breast and nonbreast regions is an important prerequisite for further image analysis. We are developing a segmentation algorithm that is fully automated and can operate independent of type of digitizing system, image orientation, and image projection. METHODS: The algorithm identifies unexposed and direct-exposure image regions and generates a border surrounding the valid breast region, which can then be used as input for further image analysis. The program was tested on 740 digitized mammograms; the segmentation results were evaluated by two expert mammographers and two medical physicists. RESULTS: In 97% of the mammograms, the segmentation results were rated as acceptable for use in computer-aided diagnostic schemes. Segmentation problems encountered in the remaining 22 images (2.9%) were most often caused by digitization artifacts or poor mammographic technique. CONCLUSION: The developed algorithm can serve as a component of an "intelligent" workstation for computer-aided diagnosis in mammography. PMID- 9419518 TI - Automated discrimination and quantification of idiopathic pulmonary fibrosis from normal lung parenchyma using generalized fractal dimensions in high-resolution computed tomography images. AB - RATIONALE AND OBJECTIVES: We computed generalized fractal dimensions for high resolution computed tomography (HRCT) images to investigate their value in the discrimination and quantification of idiopathic pulmonary fibrosis (IPF) from normal lung parenchyma. METHODS: A probability distribution that was based on the pixel value in each image was used to compute capacity, information, and higher fractal dimensions for a series of 52 HRCT slices obtained from four patients. Qualitative classification of normal, mild, moderate, and severe IPF cases was achieved by computing the following parameter: DD = D0 - 2D1 + D2, where D0, D1, and D2 represents the capacity, information, and pair correlation dimensions, respectively. A multiple linear regression analysis using morphometric quantification for the set of 52 slices was tested for all possible combinations of the parameters D0, D1, D2, and D3. The generalizability of the model was tested by predicting the extent of IPF for each patient from a regression model computed with the remaining slices in the database. RESULTS: The best regression results were obtained using the independent parameters D1 and D2 to quantify the extent of diseased lung parenchyma. The technique was tested with 48 slices from 12 new patients. The results indicated that the extent of IPF could be predicted within the confidence limits given by the regression analysis. CONCLUSION: The extent of IPF can be predicted well within the 90% confidence interval given by the model. The width of the confidence interval decreases as the number of slices used in the linear regression model increases. This operator-independent quantitative technique may be useful in the follow-up of patients with IPF. PMID- 9419519 TI - Spiral computed tomography of the liver: contrast agent pharmacokinetics and the potential for improved hepatic enhancement. AB - RATIONALE AND OBJECTIVES: We conducted a prospective study of 131 patients to evaluate the contrast agent dose-response relationship for liver spiral computed tomography (CT) and to test the hypothesis that spiral CT scanning provides greater enhancement than does dynamic CT scanning. METHODS: Patients were assigned to one of two control groups (dynamic CT) or to one of five experimental groups (spiral CT). Dynamic CT patients received 150 ml and spiral CT patients received either 75, 100, or 150 ml of diatrizoate meglumine. All groups had a monophasic injection rate of 2.5 ml/sec. Hepatic enhancement was compared among experimental and control groups. RESULTS: In the experimental groups, there was a linear dose-response relationship (p < .0001) among the enhancements achieved for the three dosages. The enhancement of the last slice of liver for the spiral CT versus dynamic CT groups receiving 150 ml was significantly greater (p = .002). Peak, first liver slice, and average liver enhancement values were higher with spiral CT scanning, but the difference was not statistically significant (power > .55). CONCLUSION: Using uniphasic injection rates and identical doses of contrast agent, spiral CT scanning has the advantage of improved enhancement of the last part of the liver to be imaged. PMID- 9419520 TI - Detectability of simulated brain activation using dual radioisotope SPECT based on size and intensity of the focal hyperactivity. AB - RATIONALE AND OBJECTIVES: Simultaneous single-photon emission computed tomography (SPECT) neuroimaging with both technetium-99m (99mTc) hexamethylpropyleneamine oxime (HMPAO) and iodine-123 (123I) N-isopropyl-iodoamphetamine is a recently introduced method with potential for assessing activation phenomena in the brain. However, there is limited information on the accuracy of the technique for detecting focal cortical sites of neuroactivation. We determined, in vitro, what levels of activation could be detected as a function of the size of the activated region. METHODS: A Lucite brain phantom was filled with both 123I and 99mTc so as to simulate both a nonactivated state (123I) along with focal sites of activation (99mTc). Simulated activations ranged from 0 to 18% in volumes of 7, 14, 20, and 27 cm3. Imaging was performed with a triple-detector gamma camera using a 10% symmetric window at 140 keV and 10% asymmetric window around 159 keV. No correction was made for gamma cross-talk. To determine whether a simulated activation was "detected," the 99mTc: 123I count ratios in the activated regions were compared by t test with ratios in nonactivated regions of similar volume. Detection sensitivities also were calculated as the fraction of the activated 99mTc: 123I ratios that were greater than the mean + 2 standard deviations of the corresponding nonactivated ratios. RESULTS: All sites of simulated activations of 10% or greater were detected. The detection sensitivity was 100% (95% confidence interval, 90-100%) for the two largest chambers with simulated activations of 13 18%. Activations in the 3-6% range, in the same-sized chambers, were detected with a limited sensitivity (67% with a confidence interval of 45-84%). In the 14 cm3 chamber, simulated activations in the 13-18% range were detected with 90% sensitivity (confidence interval, 74-98%). In general, the detection sensitivity was greater for larger chambers and higher levels of simulated activation. CONCLUSION: We conclude that the dual-radioisotope technique using triple detector SPECT systems and low-energy all-purpose (LEAP) collimators should be highly reliable for identifying focal brain activations above 13% that cover at least 14 cm3 of brain cortex. Smaller, less intense sites of activation will be detected with reduced frequency. These conclusions are based on our assessment of only the physical parameters involved in this methodology and other factors (e.g., the possibility that the relation between cerebral radiotracer concentration and regional cerebral blood flow) may affect the results obtained with patients. PMID- 9419521 TI - Cardiac hemodynamic effects of iodixanol, iopamidol, and ioxaglate following left coronary injections in anesthetized dogs. AB - RATIONALE AND OBJECTIVES: Iodixanol, a dimeric, nonionic X-ray contrast medium, has been formulated at 320 mg iodine per milliliter and supplemented with Na+, Ca2+, and Cl- to produce an osmolality that approximates that of plasma. We compared the effects of left main coronary artery injections of iodixanol, ioxaglate, and iopamidol on cardiac mechanical function in dogs. METHODS: Six mixed-breed dogs were anesthetized and prepared for recordings for electrocardiogram, aortic and left ventricular pressures, and the first derivative of left ventricular pressure, dP/dt. The test solutions and saline were injected into the left coronary artery in a randomized order. The series of four injections were repeated three times in each animal for a total of 12 injections per dog. RESULTS: Iodixanol caused significantly lower (p < .05) reduction in peak left ventricular pressure (-1.7 +/- 0.9% vs -0.7 +/- 2.0%), in diastolic aortic pressure (-1.3 +/- 1.1% vs -9 +/- 1.3%), and in left ventricular dP/dt (0.3 +/- 1.3% vs -13.2 +/- 2.4%) than did ioxaglate. Iodixanol also produced smaller cardiovascular effects than did iopamidol, but the differences were not statistically significant. Injections of both iopamidol and ioxaglate caused significant decreases from baseline parameter values; however, the changes with iodixanol were not significant. CONCLUSION: The isotonic formulation of iodixanol caused smaller cardiovascular hemodynamic effects than did iopamidol and ioxaglate and may offer increased safety in patients with severe cardiac disease. PMID- 9419522 TI - Time versus density enhancement of liver, spleen, and great vessels following rapid intravenous infusion of perflubron emulsion. AB - RATIONALE AND OBJECTIVES: We studied hepatosplenic enhancement in rhesus monkeys for 5 hr after rapid administration of perflubron (perfluorooctyl bromide [PFOB]) in an attempt to determine a clinically useful imaging window. METHODS: Five rhesus monkeys were examined using perflubron emulsion, 90% w/v perfluorochemical administered intravenously at a dose of 1.5 ml/kg and rate of 0.5 ml/sec. Helical computed tomography examination of the abdomen was obtained prior to the contrast bolus and 5 min, 30 min, 1, 2, 3, 4, and 5 hr postcontrast. Mean density of liver, spleen, and aorta was measured at each time interval. RESULTS: Significant aortic enhancement of 53 +/- 7 Hounsfield units (HU) (p < .0001) and liver enhancement of 19 +/- 4 H (p < .0004) occurred after 5 min and did not change significantly (p > .05) over 5 hr. Splenic enhancement of 35 +/- 9 HU was significant at 5 min (p < .0001) and continued to increase for 5 hr. CONCLUSION: Enhancement of the liver, blood vessels, and spleen is rapid and persists for at least 5 hr, which suggests a wider temporal window for hepatosplenic imaging with perflubron than is currently available with iodinated contrast agents. PMID- 9419523 TI - Intravenous manganese-mesoporphyrin as a magnetic resonance imaging contrast agent: an experimental model using VX-2 carcinoma in rabbits. AB - RATIONALE AND OBJECTIVES: We investigated the potential of manganese (III) mesoporphyrin (Mn-mesoporphyrin) as a hepatobiliary contrast agent for magnetic resonance (MR) imaging in rabbits given VX-2 carcinoma liver implants. METHODS: Rabbits given VX-2 carcinoma liver implants (n = 8) were imaged before and after the intravenous (i.v.) administration of 0.04 mmol/kg Mn-mesoporphyrin. MR images were correlated with gross-specimen cross-sections. The distribution of Mn in various tissues following i.v. administration of 0.04 mmol/kg Mn-mesoporphyrin was determined using atomic absorption analysis. A standard panel of serum chemistries was followed over 7 days in six rabbits following this same dose of Mn-mesoporphyrin and compared with chemistries from two control rabbits. RESULTS: I.v. administration of 0.04 mmol/kg (25 mg/kg) Mn-mesoporphyrin resulted in improvement of tumor-to-liver contrast, with enhancement of normal liver (99.7 +/ 14.7%) and the gallbladder (442 +/- 116%), but not VX-2 tumor tissue (14.8 +/- 13.9%), (n = 8, p = .05). Analysis of tissue Mn levels 100 min after i.v. Mn mesoporphyrin injection demonstrated preferential distribution of Mn to normal liver tissue (57.8 +/- 15.3 micrograms Mn/g) compared with VX-2 tumor (4.28 +/- 1.48 micrograms Mn/g). No significant change was found in the serum chemistries of six normal rabbits over a 7-day period after the i.v. administration of 0.04 mmol/kg Mn-mesoporphyrin. CONCLUSION: I.v. Mn-mesoporphyrin improved lesion-to liver contrast because of preferential distribution of Mn-mesoporphyrin to normal liver parenchyma and bile. PMID- 9419524 TI - Magnetic resonance imaging of Long-Evans cinnamon rats as a new model of hepatocellular carcinoma. AB - RATIONALE AND OBJECTIVES: Following hereditary hepatitis, Long-Evans Cinnamon (LEC) rats spontaneously develop hepatocellular carcinomas (HCC) histopathologically similar to human well-differentiated HCC. We demonstrated that LEC rats are an appropriate model of evaluating magnetic resonance (MR) imaging of well-differentiated liver tumors. METHODS: Six 23-25-month-old LEC rats were studied using liver MR imaging and histologic observation. RESULTS: Signal intensity of HCCs without cystic areas was normal or slightly high on T1 weighted images and slightly high on T2-weighted images. Histopathologically, most tumors resembled human highly or well-differentiated HCCs. CONCLUSION: The LEC rat is a good model of investigating MR imaging of well-differentiated HCC. PMID- 9419525 TI - A pressure-recording guidewire for measuring arterial transstenotic gradients: in vivo validation. AB - RATIONALE AND OBJECTIVES: We used an 0.018-inch guidewire with a pressure sensor to measure arterial transstenotic pressure gradients. Our aim was to evaluate the pressure-recording properties of this device in vivo. METHODS: Stenoses in the common carotid artery of piglets were created with external cylindrical constrictors. Pressures and pressure gradients were measured with the pressure wire and compared with reference measurements and with a 3.1-French fluid-filled catheter. RESULTS: The averages of several pressure measurements were close to the reference, but there were individual deviations. Twenty-nine percent of zero corrected systolic measurements deviated more than 5 mm Hg from the reference, and 10% deviated more than 10 mm Hg. Errors canceled out somewhat in gradient measurements. CONCLUSION: Although some measurement errors were found, the guidewire represents an important new concept for gradient measurements. The small diameter minimizes gradient augmentation caused by the measuring device lying across the stenosis. Further refinement would increase its usefulness. PMID- 9419526 TI - Percutaneous ultrasound-guided radiofrequency electrocautery ablation of prostate tissue in dogs. AB - RATIONALE AND OBJECTIVES: We investigated the feasibility of percutaneous radiofrequency (RF) electrocautery in ablation of prostate tissue in dogs. METHODS: We used six dogs in whom a specially designed needle was placed percutaneously into the prostate. RF electrocautery was applied to the needle and treatment was monitored with ultrasound. Animals were sacrificed and gross examination of the prostate and surrounding tissues was performed. Histopathologic examinations of the prostate were also performed. RESULTS: The treatment zone appeared as an elliptical echogenic focus on ultrasound that increased in size with the application of current. Gross and histopathologic correlation demonstrated that the treatment area included a central area of char with a surrounding area of coagulation. There were no deleterious effects to surrounding tissues. CONCLUSION: Our results demonstrate the feasibility of percutaneous ultrasound-guided RF electrocautery ablation of canine prostate tissue. PMID- 9419527 TI - On the generalization of the receiver operating characteristic analysis to the population of readers and cases with the jackknife method: an assessment. AB - RATIONALE AND OBJECTIVES: A new methodology that analyzes receiver operating characteristic (ROC) data sets based on jackknifing and that considers both case and reader variability has been proposed. The purpose of this investigation was to compare results using this method to those using commonly reported methodology. METHODS: ROC data sets using discrete and continuous rating scales were analyzed using the proposed jackknifing method, and results were compared to analysis of the same data sets using the paired t test. RESULTS: The two methodologies did not result in the same significance levels, and in some cases, the difference was sufficient to affect conclusions regarding comparisons of diagnostic modalities. The probability value for the jackknifing procedure is based on large sample distribution theory, and its appropriateness is unknown for sample sizes used in practice. Also, the jackknifing technique was found to be sensitive to outliers resulting when data from the computer programs used to estimate area under the ROC curve failed to converge. CONCLUSION: Although the proposed methodology yields reasonable results, several fundamental and practical issues must be addressed before it can be used widely as the analytic method of choice in ROC studies comparing different imaging techniques or reading environments. PMID- 9419528 TI - Documentation of teaching for faculty promotion. PMID- 9419529 TI - The pertinence of research on visual search to radiologic practice. PMID- 9419530 TI - Computer-assisted multimedia communication of radiologic reports. PMID- 9419531 TI - Neonate with respiratory distress and diffuse skeletal abnormalities. PMID- 9419532 TI - Subspecialization. PMID- 9419533 TI - High-resolution computed tomography sampling for detection of asbestos-related lung disease. AB - RATIONALE AND OBJECTIVES: We determined whether a limited number of high resolution computed tomography (HRCT) scans will effectively screen for interstitial lung disease (ILD) in a population of individuals exposed to asbestos. METHODS: We retrospectively reviewed the computed tomography studies of 49 patients exposed to asbestos. HRCT in the supine and prone positions had been performed at specifically preselected levels. Two teams of thoracic radiologists evaluated, on separate occasions: (1) all images, (2) prone images only, and (3) a single prone image through the lung bases for the presence of diffuse ILD. RESULTS: A relatively high level of accuracy was obtained with a single prone scan. However, improvement to 95% or better was found when additional prone images were used. CONCLUSION: A screening study for ILD, in this case patients exposed to asbestos, may be performed by preselected prone HRCT images only. The ease and decreased time of performing the procedure make screening relatively large patient groups for ILD more feasible. PMID- 9419534 TI - Observer performance comparison of digital radiograph systems for stereotactic breast needle biopsy. AB - RATIONALE AND OBJECTIVES: Two digital radiograph systems for stereotactic mammography, one using a lens to couple a Lanex Regular screen to a back illuminated charge-coupled device (CCD) and one using a fiber-optic taper to couple a Min-R Regular-type screen to a front-illuminated CCD, were evaluated with respect to observer performance. METHODS: A contrast-detail phantom was imaged in a variety of equivalent exposure conditions on both systems. Six observers viewed images on a video monitor and recorded which objects were detected. RESULTS: Performance (percent correct detections) with the lens-coupled system using the Lanex Regular screen was significantly higher than with the fiber-optic-coupled system using the Min-R Regular-type screen. CONCLUSION: Differences in absorption efficiencies of phosphors used, as well as differences in design of the two cameras, can explain differences in observer detection performance. PMID- 9419535 TI - Comparison of patient reactions and diagnostic quality for hysterosalpingography using ionic and nonionic contrast media. AB - RATIONALE AND OBJECTIVES: We compared adverse reactions and image quality for hysterosalpingography (HSG) performed with ionic (diatrizoate meglumine combined with iodipamide meglumine [DM + IM]) and nonionic (iohexol) contrast media. METHODS: We performed a study of 95 patients who had HSG and were randomly selected to receive DM + IM or iohexol. Patients reported episodes of abdominal pain and other adverse reactions immediately and 24 hr after the procedure and categorized severity of symptoms on a subjective scale. Two radiologists evaluated image quality for diagnosis. RESULTS: Prevalence of abdominal pain and other reactions both immediately and 24 hr after HSG was lower in patients who received iohexol than in patients who received DM + IM. Moderate or severe abdominal pain was significantly lower in the iohexol group than in the DM + IM group (p < .05). Visualization of the uterine cavity and ampullary rugae was judged excellent with both contrast media (87% with iohexol and 92% with DM + IM). CONCLUSION: Iohexol and DM + IM are excellent contrast media for use during HSG; iohexol 300 may cause fewer episodes of more severe and prolonged abdominal pain. PMID- 9419536 TI - Magnetic resonance (MR) imaging and MR spectroscopy of nerve regeneration and target muscle energy metabolism in a model of prosthesis-guided reinnervation in rats. AB - RATIONALE AND OBJECTIVES: We monitored the regeneration of the rat sciatic nerve after its transection and the concomitant alteration in the high-energy phosphates content in the target tibialis anterior muscle. METHODS: Rat sciatic nerve was resected and the gap connected with a prosthesis of polytetrafluoroethylene. Progress of reinnervation was monitored by 1H MR imaging, whereas muscular energy metabolism was evaluated by localized 31P MR spectroscopy. RESULTS: Reconstitution between the nerve stumps was resumed 8-12 weeks postoperatively. The ratio of phosphocreatine to inorganic phosphate reached a plateau at 46% of the initial level approximately 8 weeks after the operation and recovered thereafter. Immediately after the surgery, muscular pH became slightly alkaline and returned to normal with the progress of reinnervation. CONCLUSION: Recovery of the muscular energy metabolism began after the reconnection of the severed nerve stumps. The combination of MR imaging and MR spectroscopy followed noninvasively the progress of reinnervation and muscular energy metabolism of the prosthesis-guided nerve regeneration. PMID- 9419537 TI - Electromagnetic flow measurement using a magnetic resonance imaging static field. AB - RATIONALE AND OBJECTIVES: Electromagnetic flowmeters have been used for many years as a standard method to determine blood flow in animal models. The use of a modified probe to measure electromagnetic induction in response to an external, large direct current (DC) field magnetic resonance (MR) imaging system was investigated. METHODS: Extracted sheep iliac vessels were inserted into a pumped saline circuit with a modified probe and placed into a 2.0-T MR research imaging system. Voltage readings were collected at various flow rates ranging from 100 to 400 ml/min. Actual flows were measured with a graduated cylinder. RESULTS: A correlation of .937 (p < .001) was observed between the measured voltage changes and the actual flows. Baseline drift was also linear and within specified limits. CONCLUSION: The results indicate that electromagnetic induction in a conductive fluid can be accurately measured using electrodes and an MR imaging system and that this technique provides possible opportunities for in situ flow measurements in humans. PMID- 9419538 TI - Preclinical evaluation of manganese carbonate particles for magnetic resonance imaging of the liver. AB - RATIONALE AND OBJECTIVES: We characterized the physical, biological, and imaging properties of a manganese (Mn) carbonate particle suspension, a contrast agent for hepatic magnetic resonance (MR) imaging. METHODS: Mn carbonate suspensions were produced by controlled precipitation and characterized using light microscopy, transmission electron microscopy, and in vitro relaxivity studies. Efficacy of the agent was studied in normal and tumor-bearing rats using T1 weighted MR imaging. RESULTS: Following intravenous injection of Mn carbonate particles at doses ranging from 10 to 100 mumol Mn/kg, peak hepatic contrast enhancement of approximately 35% occurred from about 125 min until the termination of the MR imaging studies that varied from 125 to 305 min. Lesion conspicuity was increased because of relative intensity differences between normal liver and tumor. Data also showed that Mn carbonate particles dissolved on delivery to the liver, allowing Mn to interact with intrahepatic macromolecular complexes to provide positive contrast enhancement. CONCLUSION: Mn carbonate particles produce significant and sustained hepatic enhancement and should improve detection of small or isointense liver lesions. PMID- 9419539 TI - Acute hemodynamic effects of intravenous bolus injection of ionic and nonionic magnetic resonance contrast media. AB - RATIONALE AND OBJECTIVES: With the development of fast magnetic resonance (MR) imaging, bolus injection of contrast media is often required. We evaluated the acute hemodynamic effects of intravenous (i.v.) bolus injection of two MR contrast media: ionic gadopentetate dimeglumine and nonionic gadoteridol. METHODS: Twenty normal rats were anesthetized, and the left ventricular pressure (LVP), right atrial pressure (RAP), peripheral arterial pressure (PAP), positive rate pressure development (dP/dt) of the left ventricle, and heart rate were continuously monitored up to 10 min after bolus injections of 0.1, 0.3, and 0.5 mmol/kg of either media. RESULTS: Bolus injections of gadopentetate dimeglumine induced temporal reduction of the blood pressure (the peak LVP was -25.0% at 0.5 mmol/kg and -13.2% at 0.3 mmol/kg). At the injected doses of 0.5 and 0.3 mmol/kg, the systolic PAP, mean PAP, and dP/dt were reduced. The lowest peak systolic LVP was observed at around 20 sec after injection. Those suppressed values returned to the control values within 120 sec. Injection of high doses of gadoteridol caused a temporal increase in the peak systolic LVP (9.2% at 0.5 mmol/kg and 7.4% at 0.3 mmol/kg), systolic PAP, and dP/dt. Approximately 15 sec after injection, the highest peak systolic LVP was observed, and within 30 sec the value normalized. With both ionic and nonionic contrast media, a dose of 0.1 mmol/kg did not cause significant changes in hemodynamics. CONCLUSION: In normal rats, bolus i.v. injection of high-dose gadopentetate dimeglumine has negative hemodynamic effects; gadoteridol produces mild positive effects. PMID- 9419540 TI - Intraarterial biocompatibility of polyethylene terephthalate self-expandable stents implanted in porcine peripheral arteries. AB - RATIONALE AND OBJECTIVES: We tested deployment feasibility and intraarterial biocompatibility of polyethylene terephthalate (PET) self-expandable vascular stents in a porcine peripheral artery model. METHODS: To assess the thrombogenicity and neointimal response to oversized PET self-expandable stents, we implanted 10 stents in porcine common iliac arteries, followed by a 6-mm balloon inflation to 6 atm. RESULTS: All pigs survived until the study termination 6 weeks after stent implantation. Control angiography revealed stent closure in three pigs. Minimal stent luminal diameter (MSLD) was measured using a semiautomated edge detection algorithm. The difference in MSLD after stent implantation and at control after 6 weeks was not significant (4.9 +/- 0.5 mm- >4.7 +/- 1.0 mm). Histopathologic examination showed signs of thrombotic occlusion and revascularization in occluded stents. In patient stents only a mild fibromuscular neointimal response was seen. CONCLUSION: PET self-expandable stents implanted in porcine iliac arteries are possibly thrombogenic but do not lead to a significant neointimal response. PMID- 9419541 TI - Renal transplantation. PMID- 9419542 TI - Merging multimodality displays. PMID- 9419543 TI - The long-term effectiveness of a radiology memorandum. AB - RATIONALE AND OBJECTIVES: This informal prospective study was designed to document the long-term effectiveness of a radiology department policy memorandum as a communication tool. A memorandum outlining the departmental policy regarding radiographic imaging of pregnant or possibly pregnant patients served as the study model. METHODS: A departmental obstetric policy memorandum was distributed to all radiology personnel, including faculty and residents. The effectiveness of the memorandum was measured by the ability of all department personnel to answer specific test questions about the policy 1 year after its distribution. RESULTS: Nine (41%) in the faculty/fellow group and seven (41%) residents gave partially correct answers about policy content. There were no totally correct policy content answers in either group. Thirty-eight (64%) of the technology staff knew the entire policy content. CONCLUSION: Staff radiologists, fellows, and residents demonstrated an unacceptable level of knowledge concerning the policy content and location. Technologists, technology students, and clerks demonstrated a more acceptable level of knowledge of the policy than did radiologists. A single memorandum without repetition or constant surveillance is not an effective communication tool to alert radiologists to the importance of understanding and implementing policy directives. PMID- 9419544 TI - A comprehensive objective-based curriculum for radiology residents. PMID- 9419545 TI - Diffuse pulmonary disease associated with hairy cell leukemia. PMID- 9419546 TI - Program quality improvement and the radiology resident. PMID- 9419547 TI - Artificial neural networks for screening patients needing emergency cranial computed tomography scans in emergency departments. AB - RATIONALE AND OBJECTIVES: We evaluated the potential for a neural network to screen candidates for emergency cranial computed tomography (CT) scans in an emergency department setting. METHODS: Data were collected from 1625 patients undergoing emergency cranial CT scanning in two different emergency departments (EDs). Singular value decomposition (SVD) was used to remap input data for network training. Data were randomly divided into six subsets, and one was reserved as a test set to analyze network performance. Five networks were then trained on data from the five remaining sets using fivefold cross-validation. Each trained network was allowed an independent vote on need for CT scanning in each case from the test set. The majority vote was used as the final prediction. A similar analysis was done on data from each individual ED. Results are compared with prior statistical studies of the same data. RESULTS: The network performed well when predicting clinical variable patterns that consistently produced negative CT scans and on patterns that were ambiguous in terms of the CT scan results. It performed poorly, however, on patterns that consistently predicted positive scans. This last finding appears to have resulted from inadequate training material. The two populations from which data were taken were shown to be distinct, but a network trained on the combined data performed as well as the networks from the individual EDs in predicting patients requiring CT scanning. Variables with the greatest contribution to the networks' prediction were consistent with those in prior statistical studies. CONCLUSION: Although preliminary in nature, neural networks show promise as a screening device for selecting patients for emergent cranial CT scanning. PMID- 9419548 TI - Classification of microcalcifications in radiographs of pathologic specimens for the diagnosis of breast cancer. AB - RATIONALE AND OBJECTIVES: Early detection of breast cancer depends on accurate classification of microcalcifications. We have developed a computer vision system that has the potential to classify microcalcifications objectively and consistently to aid radiologists in diagnosing breast cancer. METHODS: A convolution neural network (CNN) was used to classify benign and malignant microcalcifications in radiographs of pathologic specimens. Digital images were acquired by digitizing radiographs at a high resolution of 21 microns x 21 microns. RESULTS: Eighty regions of interest selected from digitized radiographs of pathologic specimens were used for training and testing of the neural network system. The CNN achieved an Az value (area under the receiver operating characteristic curve) of 0.90 in classifying clusters of microcalcifications associated with benign and malignant processes. CONCLUSION: Classification of microcalcifications in pathologic specimens for diagnosis of breast cancer was achieved at a high level in our computer vision system, which consists of high resolution digitization of mammograms and a CNN. PMID- 9419549 TI - Influence of prior radiologic information on the interpretation of radiographic examinations. AB - RATIONALE AND OBJECTIVES: We examined whether and how the provision of previous radiologic information (previous films and reports) influences the interpretation of radiographs. METHODS: We prospectively studied 35 radiologists' interpretation of 311 plain-film radiology cases in a clinical setting. A radiologist first interpreted the current radiograph with only the information given on the consultation request. Subsequently, the same radiologist received, in random order, either the previous radiographs or the previous written reports, reviewed the diagnosis, changing it when necessary, noted the recognition of new findings, and adjusted his or her degree of confidence. A third interpretation used whichever type of information was not supplied for the second. All three readings of a study were performed at the same sitting by the same radiologist. A diagnosis and degree of confidence were recorded for each reading. RESULTS: The additional information, either radiograph or written report, significantly increased the confidence of the radiologist at each stage of interpretation. The largest increase in confidence occurred whenever previous films were introduced. In group A (second reading, reports; third reading, old films), new observations were made in 17.3% of cases on the second reading and in 19.9% on the third reading. In this group, diagnoses were changed in 14% on the second reading and in 9% on the third reading. In group B (second reading, old films; third reading, old reports), new observations were made in 16.9% of cases on the second reading and 7.3% on the third reading. Diagnoses were changed in 11% on the second reading and in an additional 5% on the third reading. Most changes were toward a more specific diagnosis. CONCLUSION: Prior information significantly increased the radiologists' confidence, facilitated new observations, and allowed more specific diagnoses. Prior radiographs were more valuable than reports in some respects. PMID- 9419550 TI - Evaluation of meniscal tears: fast spin-echo versus conventional spin-echo magnetic resonance imaging. AB - RATIONALE AND OBJECTIVES: We compared the performance of fast spin-echo (FSE) with conventional spin-echo (CSE) magnetic resonance (MR) imaging sequences in the detection of meniscal tears. METHODS: Seventy-three patients underwent MR examination of the knee for suspected internal derangement. Each patient was scanned with a CSE sequence and one of two FSE sequences. The primary difference between the two FSE sequences consisted of the echo train length (ETL). Thirty seven patients (group 1) were scanned with the FSE I sequence (ETL = 8), and 36 patients (group 2) were scanned with the FSE II sequence (ETL = 4). Menisci were graded as torn or not torn on the basis of their MR appearance. The sequences were compared with each other and with the surgical findings in 31 patients who underwent arthroscopy. RESULTS: In group 1, there was a significant discrepancy between the CSE and FSE I techniques (p = .006). The FSE I sequence detected only 11 of 19 surgically proven torn menisci as opposed to 18 of 19 detected with the CSE sequence. The FSE II sequence performed significantly better in group 2, with an accuracy equal to that of the CSE sequence. CONCLUSION: FSE sequences are extremely technique dependent with regard to detecting meniscal tears and should not replace CSE sequences in this setting until further studies are performed to optimize the technique. PMID- 9419551 TI - On the perception of the left thoracic paraspinal line. AB - RATIONALE AND OBJECTIVES: We investigated whether the left thoracic paraspinal line represents an actual change in optical density corresponding to the posterior pleural reflections or whether it represents a Mach band effect. METHODS: Using photodensitometry, we obtained averaged light intensity tracings across the left thoracic paraspinal line in 14 normal digitized anteroposterior (AP) thoracic spine radiographs. We also performed a mathematical simulation of relative brightness perception by using the contrast sensitivity function of the human visual system (assumed to represent the modulation transfer function) to filter the light intensity profiles by fast Fourier transformation techniques. RESULTS: Results from 14 normal AP thoracic spine radiographs showed no significant increase in light transmitted over the perceived location of the paraspinal line. A simple linear systems analysis of the behavior of the visual system predicted a Mach band effect corresponding in location to the observed paraspinal line. CONCLUSION: The spatial pattern of optical densities in the region of the left thoracic paraspinal line rather than the tissue composition is the crucial factor responsible for the perception of the paraspinal line. PMID- 9419552 TI - Radiologic diagnosis of cystic fibrosis in adults and children. AB - RATIONALE AND OBJECTIVES: Most radiologists are familiar with the classic chest radiographic findings of cystic fibrosis (CF) when these occur in children. We hypothesized that given the same findings, a diagnosis of CF would be less likely to be considered in an adult than in a child. METHODS: We compiled 30 pediatric and 28 adult CF chest radiographs and obtained two independent readings on each by different general radiologists among the eight who volunteered to participate as they performed their daily clinical work. The cases were presented to the readers so that they did not know which radiographs were part of the study. The association between the correct diagnosis of CF and whether the patient was an adult or a child was assessed using odds ratios and logistic regression, so that Brasfield score, Schwachman-Kulczycki score, and the patient's sex could also be considered as predictive of correct diagnosis. RESULTS: In 67% of the pediatric cases, at least one of the radiologists considered CF as a possible diagnosis, whereas they considered CF a possibility in only 50% of the adults. Both radiologists suggested the correct diagnosis in 40% of pediatric cases and only 14% of adult cases (p < .05). CONCLUSION: Because the radiographic findings were similar in the two groups of patients according to severity groupings, we believe CF was less commonly considered in the adult patient because of the traditional belief that CF is a childhood disease. PMID- 9419553 TI - Effect of contrast concentration on abdominal enhancement in the rabbit: spiral computed tomography evaluation. AB - RATIONALE AND OBJECTIVES: We examined the effect of varying the concentration of a nonionic iodinated contrast agent on hepatic and abdominal vascular enhancement using spiral computed tomography (CT) scanning. METHODS: Spiral CT scans of 10 New Zealand male rabbits were obtained after intravenous injection of 240 mg I/ml iohexol and 350 mg I/ml iohexol injected at a rate of 2 ml/sec. Each animal was studied at both concentrations using four different contrast volumes: 2.3, 1.7, 1.1, and 0.57 ml/kg. Enhancement values of the aorta, hepatic veins, inferior vena cava, and hepatic parenchyma were measured using a region-of-interest cursor. RESULTS: At all contrast doses, equal volumes of contrast iohexol-350 resulted in statistically higher hepatic (p < or = .003, paired Student's t test) as well as vascular (p < .04) enhancement compared with iohexol-240. The slopes and intercepts of the enhancement curves for iohexol-350 and iohexol-240 were not statistically different (p >> .05) when enhancement was plotted as a function of total grams of iodine administered. CONCLUSION: With spiral CT scanning, appropriate contrast doses and relative costs per dose of iohexol should be considered on the basis of total iodine load administered rather than total volume administered. PMID- 9419554 TI - Hepatic transport of the magnetic resonance imaging contrast agent gadobenate dimeglumine in the rat. AB - RATIONALE AND OBJECTIVES: Gadobenate dimeglumine is a new octadentate gadolinium (III) complex salified with meglumine. The compound is currently under evaluation as an intravenously administered paramagnetic contrast agent for magnetic resonance (MR) imaging. We investigated the mechanisms involved in the biliary excretion of gadobenate ion, the contrast-effective ion in gadobenate dimeglumine. METHODS: Biliary and urinary excretion of gadobenate ion injected intravenously to rats at 0.25 mmol/kg was studied following pretreatment with bromosulfophthalein (BSP) disodium salt, sodium taurocholate (TC), or oxyphenonium bromide (OP) and at various times after common bile duct ligation. Gadobenate ion was assayed by high-pressure liquid chromatography in bile and urine. Plasma bilirubin levels after duct ligation were measured by colorimetric assay. RESULTS: The 90-min excretion of gadobenate ion into bile accounted for 35.5 +/- 3.7% and excretion into urine for 45.7 +/- 3.5% of the injected dose (mean +/- SD). Pretreatment with BSP reduced recovery of the compound in bile to less than 1% and increased urinary excretion to 65.6 +/- 4.7%. Gadobenate dimeglumine had a substantial choleretic effect that was completely abolished by pretreatment with BSP. Pretreatment with TC and OP did not change the biliary or urinary excretion of gadobenate ion. Surgical cholestasis led to a massive increase in plasma bilirubin levels from 3.9 +/- 2.2 (day of surgery) to 129 +/- 37 mumol/L (4 days after common bile duct ligature) and decreased 6-hr cumulative biliary excretion of gadobenate ion from 45 +/- 16% to 5.3 +/- 4.2% of the injected dose. Urinary excretion increased correspondingly from 49 +/- 15% to 83 +/- 12%. CONCLUSION: The transport of gadobenate ion from plasma to bile occurs in the rat mainly through the BSP/bilirubin transport systems. PMID- 9419555 TI - Effect of contrast media on in vitro bleeding time: assessment by a hollow fiber instrument. AB - RATIONALE AND OBJECTIVES: We used a global screening device that operates under physiologic flow conditions to monitor the effects of ionic and nonionic contrast media (CM) on hemostasis. METHODS: This flow dynamic technique perfuses unanticoagulated whole blood through a hollow fiber. A leak in the fiber is created by a precision needle, and the resulting pressure fluctuations within the fiber are monitored to examine the ability of the hemostatic system to close the leak by forming a stable platelet plug. RESULTS: Both ionic and nonionic CM (25% CM/blood, v/v) were shown to lengthen the mean in vitro bleeding times (IVBTs) compared with normal blood. Ionic CM (ioxaglate and diatrizoate) consistently produced IVBTs longer than 30 min. The nonionic CM iopamidol, iohexol, and ioversol gave mean IVBTs of 16.43, 17.63, and 19.84 min, respectively. CONCLUSION: Of the three nonionic CM tested, iopamidol had the greatest probability (31%) of producing an IVBT in the normal range, with probabilities of 5% and 7% for iohexol and ioversol, respectively. Thus, iopamidol offered the least anticoagulant effect among the ionic and nonionic CM we studied. PMID- 9419556 TI - Controversial aspects of the current liver donor allocation system for liver transplantation. PMID- 9419557 TI - Lesion detection in structured noise. PMID- 9419558 TI - Current status of residency programs: survey of program directors. AB - RATIONALE AND OBJECTIVES: This survey was compiled to provide current data on the structure and content of radiology residency programs and the role of the residency program director. METHODS: A survey, created in electronic form and on paper, was distributed to all United States academic, private, and military radiology residency program directors. RESULTS: Of the 202 survey forms distributed, 168 (83%) were completed and returned. Eighty percent of respondents support national curriculum guidelines, but most do not favor national curriculum requirements. About half (53%) of programs spend 6 months in "view box" nuclear medicine; others provide some experience by lectures and on-call time. Most programs (60%) relieve residents of some call during the senior year. Some programs (28%) allow seniors time away from clinical duties to study for the oral board exam, and 17% permit time away to study for the written boards. Seventy eight percent of programs have had a "problem resident," and 47% have asked a resident to leave the program. Directors' most frequently expressed concern was threat of diminished residency numbers due to decreased funding. PMID- 9419559 TI - Painless scrotal mass in a child. PMID- 9419560 TI - The transition. PMID- 9419561 TI - Selection of processing algorithms for digital image compression: a rank-order study. AB - RATIONALE AND OBJECTIVES: We investigated non-receiver operating characteristic (non-ROC) methods for the selection of processing algorithms for digital image compression. METHODS: We performed a multipoint, rank-order study with 20 posteroanterior chest images, each processed using four different algorithms. Seven radiologists reviewed these alongside the digitized noncompressed image. Observers were forced to rank order the similarity and/or difference of the processed images to the nonprocessed image in each case. RESULTS: A two-way analysis of variance of the rankings was statistically significant (p = .025), indicating that one processing scheme yielded images that were clearly perceived as the most similar to the nonprocessed images. The selected processing scheme was not the one that yielded the lowest quantitative difference from the nonprocessed images as measured by root mean square error. CONCLUSION: Non-ROC study designs that are highly sensitive to small differences among similar images can be used to select processing algorithms. PMID- 9419562 TI - Detection of degradation of magnetic resonance (MR) images: comparison of an automated MR image-quality analysis system with trained human observers. AB - RATIONALE AND OBJECTIVES: The perceived need for magnetic resonance (MR) imaging quality control (QC) is occasionally minimized on the assumption that significant errors will be detected by the users. To evaluate the validity of this assumption, we compared the sensitivity of a test object and automated image analysis system for MR imaging QC with the sensitivity of trained human observers by evaluating images that were intentionally degraded. METHODS: Parameters for imaging the test object and normal human volunteers were set to values that decreased the signal-to-noise ratio (SNR), caused distortion, and increased the slice thickness and separation. RESULTS: The human observers were able to detect a 6-13% reduction in the SNR and distortions of more than 15% in human images. They were unable to identify 40% increases in the slice thickness. Automated analysis of test object images was able to detect all image degradations at the minimum levels applied. CONCLUSION: The poor sensitivity of the human observers indicated that degradation, especially spatial measurements, could be significantly in error before being detected through visual analysis of clinical images. These errors would be detected by automated analysis of the test object used. Further investigation is needed to better define the accuracy with which quantitative image-quality analysis predicts the effects of degraded image quality on the ability of human observers to detect subtle abnormalities in clinical images. PMID- 9419563 TI - Cascade of metastatic colorectal carcinoma from the liver to the anterior diaphragmatic lymph nodes. AB - RATIONALE AND OBJECTIVES: Metastases of colon carcinoma from the liver to porta hepatis and celiac axis lymph nodes constitute a contraindication to hepatic metastatic resection. Our objective was to determine the frequency of anterior diaphragmatic lymph node (ADLN) enlargement, another efferent pathway of hepatic lymphatic drainage, in patients with colon carcinoma. METHODS: Abdominal computed tomography scans from 50 patients with colon carcinoma in whom hepatic metastases were either present (n = 25) or absent (n = 25) were reviewed. ADLNs greater than or equal to 5 mm were considered enlarged. RESULTS: Thirteen of 25 patients with hepatic metastases had ADLNs greater than or equal to 5 mm; three of 25 patients without hepatic metastases had ADLNs greater than or equal to 5 mm. The difference was statistically significant (p = .002). CONCLUSION: Metastases of colon carcinoma from the liver to the ADLNs probably are not rare. ADLN involvement would obviate hepatic resection. The ADLNs should be assessed preoperatively in surgical candidates with hepatic metastases of colon carcinoma. PMID- 9419564 TI - Intraarterial infusion of dibutyryl cyclic adenosine monophosphate plus mitomycin C for unresectable hepatocellular carcinoma: long-term survival and response to tumor growth inhibition. AB - RATIONALE AND OBJECTIVES: Dibutyryl cyclic adenosine 3',5'-monophosphate (dBcAMP) has the capacity to promote morphologic differentiation and to inhibit tumor growth in vitro, but it has not been well researched in the clinical setting. In this study we examined the effects of intraarterial infusion of dBcAMP plus mitomycin C (MMC) on long-term survival and growth inhibition of tumors. METHODS: Thirty-one previously untreated patients with unresectable hepatocellular carcinoma (HCC) received intraarterial infusion of dBcAMP plus MMC. According to the International Union Against Cancer staging system, three patients had stage T3, eight had stage T4a, and 20 had stage T4b cancer. Growth inhibition was defined as no computed tomography (CT) scan evidence of increase in tumor diameter for at least 6 months after treatment. RESULTS: In all 31 patients with HCC, the cumulative survival rate was 34% at 1 year, 14% at 3 years, and 9% at 5 years. The median survival was 5.0 months, with the longest survival period being 92 months. Among 21 patients in whom a tumor response could be evaluated on the basis of follow-up CT studies, two had complete regression of their primary tumors. Overall, the response rate was 43% (9 of 21). Among the 12 stage T4b patients who had HCC that included the main trunk and its major branches of the portal vein, portal thrombi had disappeared in four (33%). Among eight patients who survived more than 1 year and were evaluated for tumor response using follow up CT scan studies, six (75%) had growth inhibition of tumor. CONCLUSION: Chemotherapy combined with dBcAMP and MMC showed a favorable response in approximately one third of 31 patients who had unresectable HCC. Because of our results, combined therapy should be strongly considered in the treatment of patients with HCC including occlusion of the main portal vein. PMID- 9419565 TI - Evaluation of experimentally induced renal hypoperfusion using iron oxide particles and fast magnetic resonance imaging. AB - RATIONALE AND OBJECTIVES: Renal perfusion can be evaluated with first-pass study of superparamagnetic iron oxide particles (SPIO). We applied this technique to a unilateral renal hypoperfusion model in rabbits. METHODS: Turbo fast low-angle shot sequences (acquisition time = 440 msec), after bolus injection of SPIO (100 140 mumol/kg iron), were performed in two control groups (n = 5 in each) and one group (n = 5) with a left renal blood flow reduction caused by a surgical interrenal aortic ligature (140 mumol/kg iron). Qualitative and quantitative analysis using relative blood volume (rRBV), relative blood flow (rRBF), and mean transit time (MTT) were performed. RESULTS: Signal changes were symmetric in control groups without significant differences between the kidneys. The experimental group showed a significantly delayed and less pronounced maximal reduction of signal related to a significantly decreased rRBF and increased rRBV and MTT in the left kidney (p < .05). CONCLUSION: This study shows the effectiveness of a dynamic magnetic resonance study using SPIO to detect unilateral kidney perfusion reduction. PMID- 9419566 TI - Effect of manganese dipyridoxal diphosphate on liver magnetic resonance imaging and serum bilirubin in rats with removable biliary obstruction. AB - RATIONALE AND OBJECTIVES: It is known that manganese dipyridoxal diphosphate (Mn DPDP) causes persisting liver enhancement in cholestatic rats, that free Mn++ plus bilirubin induces intrahepatic cholestasis, and that free Mn++ is released in vivo after Mn-DPDP injection. Hence, there is a concern about potential secondary intrahepatic cholestasis in patients who have biliary obstruction. In this study, we further investigated this issue. METHODS: Removable total biliary obstruction (RTBO) was induced in 12 rats. Six of them (group A) received Mn-DPDP (25 mumol/kg). The others (group B) served as control animals. The data from serial magnetic resonance imaging and serum bilirubin tests were compared. RESULTS: Without Mn-DPDP, a minimal increase of the liver intensity was observed in both groups because of cholestasis. In group A, the intensity of the liver was strongly enhanced with Mn-DPDP but normalized within 48 hr after removal of the obstruction. In both groups, total bilirubin levels increased up to 131.67 mumol/l 2 days after RTBO but rapidly decreased within 4 hr and almost normalized within 24 hr after removal of the obstruction, suggesting a lack of Mn-DPDP influence on the bilirubin level. CONCLUSION: We found that Mn-DPDP did not cause secondary intrahepatic cholestasis. Retained Mn++ is likely eliminated after restoration of bile flow. These results indicate that Mn-DPDP can be used in patients who have obstructive jaundice as long as it is followed by successful bile drainage. PMID- 9419567 TI - Effects of iodinated contrast media on pulmonary airway resistance in anesthetized guinea pigs. AB - RATIONALE AND OBJECTIVES: Bronchospasm is occasionally observed following iodinated X-ray contrast medium administration. We performed an in vivo study in guinea pigs to investigate the effects of a number of iodinated contrast media on pulmonary airway resistance and the mechanisms underlying the potential bronchoconstrictor effect. METHODS: The contrast media studied were the pharmaceutical formulations of iomeprol (400 mg I/ml), iopamidol (370 mg I/ml), and iohexol (350 mg I/ml), which are nonionic, triiodinated contrast media; diatrizoate (370 mg I/ml), an ionic, triiodinated contrast medium; iotrolan (300 mg I/ml), a nonionic, hexaiodinated contrast medium; and iocarmate (280 mg I/ml) and ioxaglate (320 mg I/ml), which are both hexaiodinated and ionic contrast media. Each contrast medium was administered intravenously at 2 g I/kg. Changes in pulmonary airway resistance were evaluated by measuring intratracheal pressure at the moment of maximum insufflation, or maximal insufflation pressure (MIP), in anesthetized guinea pigs submitted to forced ventilation. RESULTS: All contrast media except ioxaglate caused mean increases of MIP of no more than 20%. By contrast, ioxaglate caused a marked bronchoconstrictor effect, increasing MIP by 242% +/- 46%. Of the drugs tested for antagonistic action on this increase in MIP, salbutamol inhibited almost completely the increase in MIP for the first 40 min posttreatment. Similarly, lysine acetylsalicylate and indomethacin consistently reduced MIP after contrast media administration to levels only 30% and 14% above those of baseline precontrast media, respectively. Promethazine had only a minor inhibitory effect, and the response to prednisolone varied. CONCLUSION: There was no apparent relationship between the size of the increase in airway resistance and the charge or molecular weight of the contrast agent molecule or the pharmaceutical formulation. The increase induced by ioxaglate must be attributed to inherent molecular toxicity mediated through a direct action on the production of bradykinin and/or the prostanoid products of the cyclooxygenase pathway, rather than through a direct action on the release of histamine. PMID- 9419568 TI - Biodistribution and excretion of 153Gd-labeled gadolinium ethoxybenzyl diethylenetriamine pentaacetic acid following repeated intravenous administration to rats. AB - RATIONALE AND OBJECTIVES: We investigated the distribution of radioactivity in tissues and organs and its disappearance following repeated intravenous (i.v.) administration of 153Gd-labeled gadolinium ethoxybenzyl diethylenetriamine pentaacetic acid (Gd-EOB-DTPA) to rats. METHODS: A high total dose of 250 mumol/kg of 153Gd-labeled Gd-EOB-DTPA (a daily injection of 50 mumol/kg is equivalent to 92.5 kBq per animal on 5 consecutive days) was given to conscious rats by fast i.v. injection via a tail vein. The organ distribution and the body elimination into urine and feces were investigated at time points 3, 7, 14, and 21 days following the last injection; five animals were used at each time point. All samples were measured by gamma counting of 153Gd over a 10-min period. RESULTS: The radioactivity quickly disappeared from the body, mostly through feces. In the liver, no radioactivity could be detected at 3 days postinjection. At 21 days postinjection, only 0.002% of the gadolinium injected was detected, the vast majority (approximately 95%) of which was found in the kidneys. CONCLUSION: After repeated i.v. administration of a total dose of 250 mumol/kg of 153Gd-labeled Gd-EOB-DTPA, the elimination from the body was found to be 99.998% complete. Only negligible long-term retention of radioactive gadolinium was observed despite the relatively high dose injected. No perceptible evidence for decomplexation of Gd-EOB-DTPA could be found. PMID- 9419569 TI - Magnetic resonance imaging of Achilles tendon xanthomas using a fat-water discrimination technique at 0.1 T. AB - RATIONALE AND OBJECTIVES: Tendon xanthomas are atherosclerotic plaque like collections of lipids that develop with age in the Achilles tendons of patients having familial hypercholesterolemia (FH). We tested a fat-water discrimination technique for imaging Achilles tendon xanthomatosis. METHODS: We used the spin warp imaging technique optimized for low-field magnetic resonance (MR) imaging to obtain separate fat and water images of the Achilles tendon. Seven patients with FH, two patients with normal tendons, and three patients with other tendon pathology were studied. RESULTS: Normal tendons showed an intensity near or equal to that of the noisy background in all images. Compared with the background, the intensity of the FH tendons was approximately fivefold greater in magnitude images, fourfold in fat images, and 10-fold in water images. CONCLUSION: The method sensitively detected even subtle xanthomatosis in young patients, but differentiation of xanthomas from other pathologic lesions was possible with this method only when the tendons were significantly thickened. PMID- 9419570 TI - Disorders of lymph flow. AB - Disturbances in blood capillary exchange of fluid, macromolecules, and cells across intact and abnormal microvessels and deranged lymphatic transport are integral, interacting components in disorders of tissue swelling. Lymphedema or low-output failure of the lymph circulation is often indolent for many years before lymphatic insufficiency (failure) and tissue swelling emerge and persist. Superimposed occult or overt infection (lymphangitis) are probably major contributors to progressive limb deformity (elephantiasis). Long-standing lymphedema is characterized by trapping in the skin and subcutaneous tissue of fluid, extravasated plasma proteins, and other macromolecules: impaired immune cell trafficking; abnormal processing of autologous and foreign antigens; heightened susceptibility to superimposed infection; local immunodysregulation; defective lymphatic (lymphangion) propulsion from an imbalance of mediators regulating vasomotion; soft-tissue overgrowth; scarring and hypertrophy; and exuberant angiogenesis occasionally culminating in vascular tumors (Fig. 8). In contrast to the blood circulation, where flow depends primarily on the propulsive force of the myocardium, lymph propulsion depends predominately on intrinsic truncal contraction, a phylogenetic vestige of amphibian lymph hearts. Whereas venous "plasma" flows rapidly (2-3 l/min) against low vascular resistance, lymph flows slowly (1-2 ml/min) against high vascular resistance. On occasion, impaired transport of intestinal lymph may be associated with reflux and accumulation and leakage of intestinal chyle in a swollen leg. Although the term "lymphedema" is usually reserved for extremity swelling, the pathogenesis of a wide variety of visceral disorders also may be traceable to defective tissue fluid and macromolecular circulation and impaired cell trafficking of lymphocytes and macrophages. Thus, lymph stasis, with impaired tissue fluid flow, underlies or complicates an indolent subclinical course with a long latent period and sporadic episodes of lymphangitis, which culminates in intense scarring. Examples are pulmonary fibrosis (e.g., pneumoconiosis), regional enteritis, retroperitoneal fibrosis, and perhaps chronic pancreatitis and cirrhosis of the liver. Transdifferentiation and ultimately transformation of endothelial and other vascular accessory cells during lymph stasis also may be pivotal to a wide range of dysplastic and neoplastic vascular disorders, including Stewart-Treves angiosarcoma, AIDS-associated Kaposi's sarcoma, and lymphangitic metastatic carcinomatosis. Lymphscintigraphy has now replaced conventional lymphography as the procedure of choice to corroborate the diagnosis of peripheral lymphedema, whereas MR imaging using paramagnetic and superparamagnetic contrast agents has the potential to yield huge dividends in furthering understanding of a variety of enigmatic edematous states, including lymphedema. Not only are better explanations and insights into swelling disorders likely to be forthcoming, but, equally important, these new, safe, noninvasive imaging techniques can and should be used to monitor the evolution and document the efficacy of commonly advocated operations and nonoperative remedies for defective lymph transport and function. PMID- 9419571 TI - Three-dimensional imaging techniques: a current perspective. PMID- 9419572 TI - Faculty contracting for the technical management of an academic radiology department: a case study. PMID- 9419573 TI - Increasing medical student interaction: the radiology passport. PMID- 9419574 TI - Progressive painful bowing of the right leg. PMID- 9419575 TI - Being a radiologist. PMID- 9419576 TI - Clinical research in a time of change. PMID- 9419577 TI - In vivo characterization of cytotoxic intracellular edema by multicomponent analysis of transverse magnetization decay curves. AB - RATIONALE AND OBJECTIVES: We investigated the multicompartmental nature of T2 decay in a specific white matter edema model. METHODS: Triethyltin (TET) intoxication was produced in six male New Zealand White rabbits. Images were obtained over the 23-day study duration using a 64-echo Carr-Purcell-Meiboom-Gill (CPMG) sequence (repetition time = 3000 msec, echo time = 20 msec). T2 decay curves were extracted from 0.7 x 0.7 x 3.0 mm3 voxels in the corpus callosum and contiguous white matter tracts, cortex, thalamic nuclei, hypothalamic nuclei, and the masseter muscles. The curves were fit with biexponential functions. RESULTS: Increased signal intensity in the corpus callosum was evident 2-3 days after the first TET injection. At this time, a substantial slowly relaxing component appeared in the decay curves of the corpus callosum and, to a lesser extent, in the thalamus and hypothalamus. Changes in the rabbits' body weight, general physical condition, and neurologic state paralleled the growth and regression of the second, slowly relaxing component. CONCLUSION: The appearance and regression of a slowly decaying second component in the T2 decay curve is consistent with the formation and shrink-age of intracellular vesicles in the intramyelin sheaths of central white matter. PMID- 9419578 TI - Renal resistive index in experimental partial and complete ureteral obstruction. AB - RATIONALE AND OBJECTIVES: Recent clinical work suggests that the Doppler resistive index (RI) may be useful in distinguishing obstructive from nonobstructive hydronephrosis. We evaluated the usefulness of the RI in a rabbit model of hydronephrosis. METHODS: Unilateral partial ureteral obstruction was produced in nine rabbits and complete obstruction in another nine. Three sham operations were performed, and these animals served as control subjects. The RI was measured in all kidneys before and 6 hr after surgery and on days 1, 4, and 7 postoperatively. The RI and the difference in RI (delta RI) between the obstructed and normal kidney were evaluated over time using a two-way analysis of variance. The intravenous urography and Whitaker tests served as gold standards. RESULTS: Hydronephrosis was observed on sonograms in all obstructed kidneys. Comparing groups, there was no significant difference in mean RI or delta RI between the three groups at any time point. Looking at individual groups over time, there was no significant change in mean delta RI, whereas the change in mean RI was significantly elevated above baseline only in the complete obstruction group at 6 hr (p = .002) and on days 4 (p = .008) and 7 (p = .006). In evaluating varying thresholds of RI and delta RI, we could not consistently discriminate between normal and obstructed kidneys. CONCLUSION: Although complete obstruction caused a significant increase in RI, partial obstruction failed to do so. RI and delta RI values proved to be insensitive predictors of obstruction in this rabbit model. PMID- 9419579 TI - Reaction of the aortic wall to six metallic stent materials. AB - RATIONALE AND OBJECTIVES: We investigated the effects of various metallic stents on the aortic wall. METHODS: The wires of Gianturco-type expandable metallic stents were plated with gold, silver, or copper or coated with Teflon or silicone. Stents were inserted into the aortas of 15 adult mongrel dogs. The time course of radiologic, macroscopic, and histologic changes in the aorta at the site of the stent was investigated at 1, 2, and 4 weeks after implantation. RESULTS: The gold-plated stent appeared to produce fewer macroscopic and histopathologic changes in the aorta than the other types of stents. The neointima was thinnest with gold (83.9 +/- 40.3 microns), followed by stainless steel (103.6 +/- 57.0 microns), Teflon (115.0 +/- 30.2 microns), silicone (209.6 +/- 25.9 microns), silver (228.6 +/- 33.8 microns), and copper (unmeasurable). With the copper-plated stent, the aorta suffered severe erosion of the vessel wall, marked thrombus formation, and aortic rupture. CONCLUSION: Gold is a useful intravascular material because it reacts only minimally with the vessel wall. PMID- 9419580 TI - A theoretical model using mechanical principles to quantify the physical principles of balloon dilatation. AB - RATIONALE AND OBJECTIVES: Balloon dilatation is a mechanical form of controlled injury used to alleviate vascular stenoses. Several factors influence successful angioplasty. Few mechanical models exist to illustrate the physical principles of balloon dilatation. METHODS: We used mechanical analysis of membrane stresses, along with Laplace's law, to determine a relation between balloon inflation and dilating pressures exerted by balloons in stenoses of varying severity, length, and eccentricity. The balloons were assumed to be perfectly inelastic and flexible. We also examined the resultant stresses in the lesion wall of concentric and eccentric stenoses from exertion of dilating pressures. RESULTS: Dilating pressures depend directly on maximal balloon inflation pressure and balloon diameter. Short, focal stenoses experience greater dilating pressures, which often are several multiples of the inflation pressure, than similarly narrowed longer lesions. CONCLUSION: Dilating pressures depend on inflation pressure, balloon diameter, and lesion severity. PMID- 9419581 TI - Neutron capture therapy with gadopentetate dimeglumine: experiments on tumor bearing rats. AB - RATIONALE AND OBJECTIVES: The therapeutic effect of neutron capture therapy with the gadolinium (Gd) complex gadopentetate dimeglumine was studied in vivo in rats using a beam of thermal neutrons. METHODS: Rats with Jensen sarcomas 10-15 mm in diameter in their right thigh were irradiated with a thermal neutron beam that had fluences of 3.6 x 10(11) (20 min) or 5.4 x 10(11) n/cm2 (30 min) in the absence and presence of 5,500 or 13,750 ppm gadopentetate dimeglumine. Gadopentetate dimeglumine was administered directly into the tumor prior to neutron irradiation. Four groups of rats were studied: two groups of nonirradiated controls (Gd-n- and Gd+n-) and two irradiated groups (Gd-n+ and Gd+n+). In the follow-up period, we measured the subjects' clinical status and tumor size as a function of time postirradiation. RESULTS: In both control groups (Gd-n- and Gd+n-), the tumor progressively grew. Pure irradiation by thermal neutrons in the Gd-n+ group resulted in a transient inhibition of tumor growth with total regressions of 15%. Intratumoral administration of 13,750 ppm gadolinium per gram of tumor and subsequent neutron irradiation (the Gd+n+ group; fluence = 3.6.10(11) n/cm2) significantly increased the tumoricidal effects (i.e., decrease of tumor growth up to a complete regression of the tumors in about 80%). Treatment-specific differences between the groups were confirmed by histologic observations. CONCLUSION: The intratumoral administration of the hydrophilic magnetic resonance imaging contrast medium gadopentetate dimeglumine prior to irradiation with thermal neutrons leads to a therapeutic gain (i.e., reduction) on experimental Jensen sarcomas. PMID- 9419582 TI - Tissue ablation with radiofrequency: effect of probe size, gauge, duration, and temperature on lesion volume. AB - RATIONALE AND OBJECTIVES: We evaluated the parameters affecting the size and distribution of thermal tissue damage produced by radiofrequency electrodes. METHODS: Thermal lesions were produced by electrodes connected to a radiofrequency generator in specimens of liver (n = 143) and muscle (n = 20). Various combinations of probe tip exposure (0.5-8 cm), gauge (12-24 gauge), duration of treatment (0.5-12 min), and temperature (80-90 degrees C) were studied. The resulting volumes of tissue coagulation were measured and compared. RESULTS: Lesions equal to or less than 1.6 cm in diameter were symmetrically distributed around the electrode. Lesion diameter (but not length) increased with probe gauge and duration of treatment to a maximum of 6 min. However, lesions with mean diameters larger than 1.6 cm could not be produced using a single probe with any technique. Lesion length correlated with probe tip exposure from 1 to 8 cm (r2 = .996). Over the limited range investigated, increased temperature had minimal effects, except for tip exposures greater than 5 cm, in which larger and more uniform lesions resulted. Lesions varied equal to or less than 3 mm in diameter and equal to or less than 5 mm in length for each combination of variables. CONCLUSION: Radiofrequency ablation can accurately and reproducibly cause coagulative tissue necrosis. Necrosed tissue volume increases with length of exposed probe tip, larger probes, and sessions lasting at least 6 min. PMID- 9419583 TI - Indirect computed tomography lymphography of subdiaphragmatic lymph nodes using iodinated nanoparticles in normal dogs. AB - RATIONALE AND OBJECTIVES: We evaluated the imaging characteristics of an iodinated particulate contrast agent for indirect computed tomography (CT) lymphography of normal subdiaphragmatic lymph nodes in dogs. METHODS: Four milliliters of a 15% (wt/vol) iodinated nanoparticle suspension was injected into the gastric, colonic, rectal, or cervical submucosa, loose paraprostatic fascia, or metatarsal subcutaneous tissues in 10 healthy beagles. Endoscopic, CT, or ultrasound guidance was used when necessary to facilitate contrast agent delivery. CT and radiographic images were obtained prior to contrast administration and at 4 hr, 24 hr, and 7 days postcontrast injection. Postmortem examinations were then conducted. RESULTS: CT images showed enhancement of regional lymph nodes draining the various injection sites. The mean attenuation of opacified nodes was 678 +/- 463 Hounsfield units 24 hr after injection and remained elevated 7 days later. Lymph node opacification on CT images correlated well with the node location observed on postmortem examinations. CONCLUSION: Subdiaphragmatic lymph nodes can be effectively opacified using an iodinated nanoparticle contrast agent for indirect CT lymphography. PMID- 9419585 TI - Cardiovascular assist devices. PMID- 9419584 TI - Intraarticular tolerability and kinetics of gadolinium tetra-azacyclododecane tetraacetic acid. AB - RATIONALE AND OBJECTIVES: We assessed the tolerability and the intraarticular kinetics of gadolinium tetra-azacyclododecane tetraacetic acid (Gd-DOTA) using magnetic resonance (MR) imaging. METHODS: Twelve of 18 dogs received an intraarticular injection of Gd-DOTA solution. Pathologic examination of all joints was performed with assessment of Gd-DOTA bone absorption. Effects of Gd DOTA on chondrocyte viability and proliferation in vitro were determined using cultures of rabbit chondrocytes. Four dogs underwent MR imaging of the stifle joint before and after intraarticular injection of 0.8 ml Gd-DOTA at a concentration of 2 mmol/l (300 mOsm/l). Intraarticular kinetics of Gd-DOTA were determined from quantitative measurements using repeated sagittal spin-echo T1 weighted images. RESULTS: No microscopic changes of the joints or Gd-DOTA bone absorption were detected. No cytotoxicity for chondrocytes was observed at a concentration of 5 mmol/l, but a decreased cell count was observed at a high concentration (50 mmol/l). The intraarticular Gd-DOTA concentration decreased with time according to a logarithmic curve with an intraarticular half-life of 103-152 min (M = 127 min). CONCLUSION: Gd-DOTA is a safe intraarticular contrast agent with a long half-life in the joint cavity. PMID- 9419586 TI - Copyright law and academic radiology: rights of authors and copyright owners and reproduction of information. PMID- 9419587 TI - Magnetic source imaging: basic principles and applications in neuroradiology. PMID- 9419588 TI - Recurrent brain tumor and new pulmonary nodules. PMID- 9419589 TI - Legal system. PMID- 9419590 TI - The effect of storage on the computed tomography attenuation of gallstones. PMID- 9419591 TI - Computerized analysis of interstitial infiltrates on chest radiographs: a new scheme based on geometric pattern features and Fourier analysis. AB - RATIONALE AND OBJECTIVES: Detection of interstitial infiltrates on chest radiographs is difficult and subjective. Therefore, we developed a computerized method to provide quantitative analysis of lung texture to increase diagnostic accuracy. METHODS: Two hundred chest radiographs--100 healthy and 100 abnormal with interstitial infiltrates--were digitized using a laser scanner. They were analyzed by an automated computerized scheme that uses a combination of two methods for detection of interstitial infiltrates: a lung texture analysis based on the Fourier transform and a geometric pattern feature analysis based on filtering techniques. RESULTS: The overall sensitivity and specificity of the computerized scheme were 92% and 90%, respectively. The scheme achieved a sensitivity of 80% in subtle cases (n = 15) and 88% in cases with localized interstitial disease (n = 26), whereas the specificity remained unchanged. There was good correlation between the computer output and the radiologists' severity rating. CONCLUSION: This enhanced computerized scheme exhibits high sensitivity and specificity with a large database. PMID- 9419592 TI - Quantitative chest computed tomography as a means of predicting exercise performance in severe emphysema. AB - RATIONALE AND OBJECTIVES: We assessed the value of quantitative high-resolution computed tomography (CT) as a diagnostic and prognostic tool in smoking-related emphysema. METHODS: We performed an inception cohort study of 14 patients referred with emphysema. The diagnosis of emphysema was based on a compatible history, physical examination, chest radiograph, CT scan of the lung, and pulmonary physiologic evaluation. RESULTS: As a group, those who underwent exercise testing were hyperinflated (percentage predicted total lung capacity +/- standard error of the mean = 133 +/- 9%), and there was evidence of air trapping (percentage predicted respiratory volume = 318 +/- 31%) and airflow limitation (forced expiratory volume in 1 sec [FEV1] = 40 +/- 7%). The exercise performance of the group was severely limited (maximum achievable workload = 43 +/- 6%) and was characterized by prominent ventilatory, gas exchange, and pulmonary vascular abnormalities. The quantitative CT index was markedly elevated in all patients (76 +/- 9; n = 14; normal < 4). There were correlations between this quantitative CT index and measures of airflow limitation (FEV1 r2 = .34, p = 09; FEV1/forced vital capacity r2 = .46, p = .04) and between maximum workload achieved (r2 = .93, p = .0001) and maximum oxygen utilization (r2 = .83, p = .0007). CONCLUSION: Quantitative chest CT assessment of disease severity is correlated with the degree of airflow limitation and exercise impairment in pulmonary emphysema. PMID- 9419593 TI - Computed tomography appearance of the prostatic fossa following radical prostatectomy. AB - RATIONALE AND OBJECTIVES: We assessed the value of postsurgical computed tomography (CT) in detecting locally recurrent prostatic carcinoma and determined the most effective CT findings. METHODS: We studied 13 patients with recurrent disease (prostate specific antigen [PSA] > 0.4 ng/ml) and 11 patients with no evidence of recurrence (PSA < 0.3 ng/ml). Pelvic CT scans were independently interpreted by four readers who were unaware of patient status. Readers measured tissue volume in the prostatic fossa and evaluated each scan for the presence of six potentially discriminating criteria. We determined sensitivity and specificity and developed mean and pooled receiver operating characteristic (ROC) curves for each criterion and for overall rating. RESULTS: The respective mean ROC curves, sensitivity, and specificity for each criterion were as follows: irregular tissue margins = .50, .67, and .32; inhomogeneous tissue density = .35, .76, and .11; asymmetric residual seminal vesicles = .68, .86, and .16; fat infiltration around seminal vesicles = .67, .69, and .43; infiltration of perirectal fat = .60, .71, and .40; and margins of the levator ani = .50, .78, and .09. The overall rating of whether a scan was normal or reflected recurrent cancer was .56, .75, and .32. Mean tissue volume in the prostatic fossa was 15.01 cm3 for the positive cases and 11.06 cm3 for the negative cases (p < .05), but because of a large overlap, this difference was not practically significant. CONCLUSION: CT scanning is not an effective technique for detecting recurrent prostate malignancy. Normally, there is a moderate amount of soft tissue in the prostatic fossa postoperatively that should not be confused for malignancy. PMID- 9419594 TI - Intravascular hyperthermia: experimental study of transcatheter treatment. AB - RATIONALE AND OBJECTIVES: External hyperthermia has frequently been used to treat superficial malignant tumors. We postulated that transarterial internal hyperthermia may be effective for deeply located hypervascular tumors. We performed intravascular hyperthermia (IVH) for malignant hypervascular tumors that were transplanted into animals and evaluated the resulting histologic changes and antitumoral effects. METHODS: We designed a special catheter to heat the injected saline. Sixteen rabbits with hypervascular VX2 tumors ranging from 2 to 2.5 cm in diameter in their left hindlimb muscles served as subjects. IVH was performed via the left femoral artery (temperature = 50 degrees C) in 11 rabbits. Two sessions of IVH were performed during 1 week. Two weeks after the two sessions of IVH, the rabbits were sacrificed and their tumors and feeding arteries were resected. Five control rabbits had 37 degrees C saline infused using the same technique. RESULTS: The temperature in the central zone of the tumor increased markedly to 42.3 +/- 0.5 degrees C (mean +/- standard deviation), compared with 40.3 +/- 0.4 degrees C and 39.2 +/- 0.5 degrees C in the peripheral zones of the tumor and the surrounding tissue, respectively. Tumor sizes were calculated on angiograms as having decreased by 63 +/- 35% of their initial sizes during the 2 weeks after the two IVH sessions. However, tumor sizes increased to 171 +/- 41% in the control group (p < .01). Necrosis in the peripheral zones of the tumors in the IVH group was more severe than that in the control group (p < .05). Necrosis in the central zones of the tumors in the IVH group was slightly more severe. The surrounding tissue was not damaged. Although the femoral artery was not severely damaged, there was severe damage to the feeding arteries. CONCLUSION: IVH reduced the size of hypervascular tumors in our rabbits. The antitumoral effects are attributed to direct damage of the tumor vessels and to the effect of heat on the tumor cells. PMID- 9419595 TI - Lesion visualization by targeted computed tomography liver enhancement in dogs. AB - RATIONALE AND OBJECTIVES: We conducted a pilot study to determine the potential advantages of using liver-specific targeted computed tomography (CT) contrast agents for lesion detection. METHODS: Eight dogs had liver infarcts created by percutaneous injections of ethanol. Each dog underwent CT scans with four imaging techniques: unenhanced, intravenous contrast enhanced (IVCE), CT arterial portography (CTAP), and targeted liver enhancement with iodipamide ethylester (IDE) particles. Lesions were assessed quantitatively to determine liver-to lesion density differences and the drop in density across liver edge as a quantitative measure of edge sharpness. Expert readers subjectively analyzed data to determine lesion visibility and edge sharpness. RESULTS: Liver-to-lesion density differences were greatest with CTAP (56.4 +/- 35.5 Hounsfield units [H]) followed by IDE (41.1 +/- 7.0 H), i.v. (22.7 +/- 6.0 H), and unenhanced scans (13.6 +/- 4.1 H; ps < .05 for CTAP versus unenhanced and IDE versus unenhanced). Edges were best defined both subjectively and quantitatively on IDE-enhanced scans. CONCLUSION: Targeted liver-specific contrast agents have potential to increase lesion visibility when compared with standard i.v. contrast enhancement of the liver by increasing lesion edge definition and liver-to-lesion attenuation differences. Further work in animal tumor models, and clinical trials as agents become available, appears justified. PMID- 9419596 TI - Effect of magnetic resonance exposure combined with gadopentetate dimeglumine on chromosomes in animal specimens. AB - RATIONALE AND OBJECTIVES: We tested for the presence of a form of unstable chromosomal damage--anaphase bridges--that might result from the combined exposure to gadopentetate dimeglumine and magnetic resonance (MR) imaging in rats. METHODS: Fifty-four male Sprague-Dawley rats, along with appropriate controls, were exposed either to MR imaging alone, gadopentetate dimeglumine alone, or a combination of the two. After exposure, partial hepatectomies were performed to induce a vigorous mitotic response in the regenerating liver stump. Twenty-eight to 30 hr after partial hepatectomy, tissue specimens from regenerating liver were removed and analyzed microscopically for the presence of anaphase bridges. RESULTS: No anaphase bridges were detected in any of the animals, including those exposed to gadopentetate dimeglumine and MR imaging. CONCLUSION: Using anaphase bridge formation as an indicator, exposure to MR imaging alone, gadopentetate dimeglumine alone, or a combination of the two under the conditions used in this experiment did not cause detectable unstable chromosomal damage. PMID- 9419597 TI - A molecular receptor-binding contrast agent for magnetic resonance imaging of the liver. AB - RATIONALE AND OBJECTIVES: A gadolinium complex of polydiethylenetriamine pentaacetic acid polyneogalactosyl polylysine (Gd-DTPA-gal-PL) was developed and tested as a paramagnetic contrast agent for magnetic resonance (MR) imaging of the liver. The agent was designed for receptor-mediated uptake by the asialoglycoprotein receptor (ASGP-R), which is unique to hepatocytes and exhibits high specificity for galactose-terminated glycoconjugates. METHODS: Polylysine was alkylated with a mixed anhydride of diethylenetriamine pentaacetic acid. This product was complexed with gadolinium and N-alkylated with 3-oxopropyl-1-thio beta-D-galactopyranoside. With this reaction sequence, we prepared a gadolinium complex consisting of 2284 galactose groups and 858 chelators per polylysine having 2136 amino groups. Hepatic enhancement was tested by MR imaging of nine rats with liver-implanted mammary adenocarcinoma before and after injection of 20 x 10(-9) mol/kg Gd-DTPA858-gal2284-PL2136. The conjugate was labeled with technetium-99m and tested (1.5 x 10(-10) mol/kg) for hepatic specificity via nuclear imaging. RESULTS: Mean hepatic enhancement was 86% within 10 min and remained constant for 25 min. Hepatic relative intensity exceeded preinjection intensities by at least four times the standard deviation of the preinjection values (p < .01). The tumors, which are devoid of ASGP-R, did not exhibit significant enhancement (p > .1). The liver accumulated 90% of the technetium-99m labeled conjugate. CONCLUSION: A molecular paramagnetic ligand to the asialoglycoprotein receptor has been developed for hepatocyte-specific MR contrast enhancement. PMID- 9419598 TI - Injection of ethanolamine oleate into a segmental portal branch for pharmacologic hepatic segmentectomy in dogs. AB - RATIONALE AND OBJECTIVES: Currently available treatments for hepatocellular carcinoma are not satisfactory in terms of recurrence rates. In this study, we injected ethanolamine oleate (EO) into a portal branch in an attempt to cause necrosis of a liver segment in which hepatocellular carcinoma might be located. METHODS: Nine dogs received EO injections via a balloon catheter into a segmental portal branch of the liver. RESULTS: Immediately after injection, 80-100% of the liver cells in the EO-injected segment underwent coagulative necrosis. After 1 week, the EO-injected segment had become completely necrotic in two dogs. Only a few viable hepatocytes were still observed around the arteries and beneath the liver capsule in another dog. No pathologic changes were observed in the lungs, kidneys, or heart of any dog. There was a correlation between the EO dosages and the volume of the EO-injected liver tissue. CONCLUSION: EO injection into a portal branch results in the pharmacologic destruction of the corresponding liver segment. This procedure may be beneficial in the treatment of hepatic malignancies. PMID- 9419600 TI - Image quality, the ideal observer, and human performance of radiologic decision tasks. AB - The quality of medical images must be quantified with reference to specific diagnostic tasks. Image quality is limited by fundamental physics, engineering limitations, radiation safety concerns, and imaging time constraints (among other things). There is now a gold standard for assessing human visual decision performance: the ideal Bayesian observer. Unfortunately, there are no mathematical tools to use this gold standard for realistically complex tasks. As an alternative, one can use the optimum linear discriminator (Fisher-Hotelling) model as a silver standard while en route to clinical realism. The goal of scientists working in the area is to develop mathematical models of human observers that will help equipment designers to optimize design trade-offs for specific diagnostic tasks. The current strategy is to modify the Fisher-Hotelling model to include certain limitations of the human observer visual system. The model must be both robust enough and mathematically tractable enough to be used to predict performance for clinical classification and estimation tasks. Statistical models also must be developed that describe realistic signals (lesions and abnormalities) and the normal patient structure that is the background in which these signals must be detected or identified. PMID- 9419599 TI - A microcholangiographic study of liver disease models in rats. AB - RATIONALE AND OBJECTIVES: Rats develop hepatobiliary injury due to small bowel bacterial overgrowth (SBBO) that, at specimen, resembles cholangiography sclerosing cholangitis. To better visualize the smaller bile ducts, we used microcholangiography to determine the spectrum of biliary lesions in this and five other models of liver disease. METHODS: The models studied were as follows: (1) Surgically created jejunal, self-filling blind loops induce SBBO. (2) Intraperitoneal injection of a bacterial cell wall polymer, peptidoglycan polysaccharide (PG-PS), causes granulomatous hepatitis. (3) Intraperitoneal injection of endotoxin (lipopolysaccharide) causes sinusoidal congestion and shock. (4) Bile duct ligation induces bile duct proliferation. (5) Alpha-naphthyl isothiocyanate (ANIT) induces bile duct proliferation. (6) Carbon tetrachloride (CCl4) causes fibrosis and cirrhosis. Warmed barium sulfate, gelatin, and saline were injected in the extrahepatic bile duct. Liver slices (2 mm) underwent microradiographic techniques, and images were correlated with histology. RESULTS: Rats with SBBO had irregular and dilated extrahepatic bile ducts with thickened walls. Rats treated with endotoxin and CCl4 had normal microcholangiograms. Bile duct proliferation was identified following ANIT and bile duct ligation. Rats given PG-PS demonstrated irregular intrahepatic bile ducts. Microcholangiograms following SBBO and PG-PS showed similarities including focal ductal dilatation, narrowing, proliferation, and destruction. CONCLUSION: Various models of liver injury induce characteristic cholangiographic appearances. Microcholangiography is useful in examining biliary tract lesions and complements histology. PMID- 9419601 TI - Medical students who choose a radiology elective: career decisions, motivations, and intentions. AB - RATIONALE AND OBJECTIVES: A national survey of American medical students was conducted to determine the effects of taking a radiology elective. The survey also probed attitudes of students not planning a career in radiology relative to the interpretation of medical images independently of radiologists. METHODS: The names of all students taking an elective in radiology in 1993-1994 were solicited from all American medical schools. Eighty-one of 124 radiology elective programs responded, and 3872 questionnaires were returned to the 81 medical schools for delivery to the students. Approximately 25% of the questionnaires were returned, and data were derived from these. RESULTS: Rarely did the radiology elective influence a change (2.1%) from another specialty or primary care to radiology. Eighty-nine percent of the students sampled indicated that the elective had no effect on their decision. Of students who were undecided about their careers, 13.2% indicated that the elective changed their career choice, but 63.2% said the elective had no effect. The data revealed that there are multiple, often overlapping, reasons for a medical student not planning a career in radiology to choose a radiology elective. Most of the students (93.5%) expected to interpret radiographs or other medical images themselves and then correlate their impression with a radiologic report. However, 30% of the students planned to interpret some radiographs independently of radiologists, and another 15% expected to interpret medical images other then plain radiographs without a radiologic consultation. CONCLUSION: Our data suggest that attitudes regarding interpretation of radiographs or other medical images without involvement of a radiologist are ingrained in medical students and correspond to attitudes among practicing clinicians. There is an expectation among medical students that taking a 4-week elective is useful or even necessary to independently interpret radiographs in future clinical practice. The effect of these attitudes on the cost and quality of future medical care should engender considerable concern. A basic conceptual change in the structure and content of the radiology elective is suggested. PMID- 9419602 TI - A paradigm for computer-assisted radiology resident rotation scheduling. AB - RATIONALE AND OBJECTIVES: The scheduling of radiology residents remains a major annual undertaking of chief residents. In this article, we describe a paradigm to implement interactive computer programs to reduce the inefficiencies and inequities of planning the yearly schedule. METHODS: We used the programming language, Prolog, to develop a compact program that provides faster and more flexible performance than those reported in the literature. This interactive program stores scheduling requirements in data files separated from the control program and runs on a Macintosh computer. RESULTS: The schedule of any residency year is generated within 3-7 sec. The fast computation and query capabilities of this scheduling program have helped chief residents to identify conflicting requirements that were previously overlooked. CONCLUSION: Using our programming paradigm, we have developed a portable Prolog-based scheduling program that is quick and easy to use. PMID- 9419604 TI - Radiology opportunities in the military health care system. PMID- 9419603 TI - Lytic bone lesion in a 12-year-old. PMID- 9419605 TI - A role for nitric oxide in X-ray contrast material toxicity. AB - RATIONALE AND OBJECTIVES: We assessed the role that nitric oxide (NO) plays in contrast media (CM) toxicity, using 100% lethal dose (LD100) studies in hyperimmune Brown Norway (BN) rats. METHODS: Ninety-two BN rats and 41 Sprague Dawley (SD) rats underwent CM LD100 tail vein injections with methylglucamine iothalamate or sodium iothalamate to the point of cessation of respiration. Methylglucamine hydrochloride also was injected. The injections were accompanied by L-arginine (L-Arg) or D-arginine (D-Arg) analogues or by an H1 blocker. L-Arg analogues inhibit NO formation, and D-Arg analogues do not. RESULTS: An L-Arg analogue, but not a D-Arg analogue, increased the tolerance of BN rats (p < .005) for methylglucamine iothalamate but not for sodium iothalamate. The L-Arg analogue also protected BN rats against methylglucamine chloride injections (p < .002). H1 blockade protected BN rats against methylglucamine iothalamate (p < .0005) and methylglucamine chloride (p < .005) injections. None of these measures altered the CM tolerance of SD rats. In SD rats, injections of either methylglucamine iothalamate or sodium iothalamate along with a D-Arg analogue or normal saline were better tolerated than similar injections in BN rats (p < .01 and .002 for methylglucamine iothalamate and sodium iothalamate, respectively). In SD rats but not BN rats, sodium iothalamate was better tolerated than was methylglucamine iothalamate (p < .0005). CONCLUSION: NO appears to play a significant role in BN rats LD100 CM toxicity and has been implicated by others in the blood pressure fall characterizing some forms of antigen-induced anaphylaxis [1, 2]. The results of the current study and the literature suggest that methylglucamine-modulated release of histamine from mast cells may underlie the NO production. PMID- 9419606 TI - Magnetic resonance imaging with superparamagnetic iron oxide particles to evaluate hepatic macrophage-monocytic phagocytosis after arterial devascularization in minipigs. AB - RATIONALE AND OBJECTIVES: We examined the effects of arterial ischemia on the phagocytic activity of the hepatic macrophage-monocytic phagocytic system (MMPS). METHODS: Six minipigs were studied before and 24 hr after complete arterial devascularization of the liver. Magnetic resonance (MR) imaging was performed at 1.5 T using superparamagnetic iron oxide (SPIO) particles (18 mumol Fe/kg body weight) as an MMPS-specific contrast agent. Hepatobiliary scintigraphy, measurements of serum liver enzymes, and histology also were obtained. RESULTS: On MR imaging, the postcontrast-to-precontrast ratios of the arterially devascularized livers were significantly higher than the corresponding baseline values (p < .01). The greatest difference (52%) between the baseline and the postoperative values was observed on gradient-echo (GE) images. Scintigraphy, laboratory analyses, and histology results indicate that the MR imaging findings were probably predominantly attributable to a reduction in phagocytic activity of the hepatic MMPS. CONCLUSION: SPIO particles have already proved useful for improving detection of liver neoplasms on MR imaging, but they also may provide a novel way of evaluating the function of the hepatic MMPS in liver diseases. PMID- 9419607 TI - Enhancement of computed tomography liver contrast using iomeprol-containing liposomes and detection of small liver tumors in rats. AB - RATIONALE AND OBJECTIVES: We evaluated iomeprol-containing liposomes (Lipiom), a new contrast medium for computed tomography (CT) liver scanning, in an animal model of chemically induced hepatocellular carcinomas and other liver tumors in rats. METHODS: Liver tumors were induced by administration of carcinogens to rats, either 0.55% (w/w) 1'-hydroxysafrole in the diet or induction by 3'-methyl 4-diethylaminoazobenzene followed by promotion with carbon tetrachloride. CT scanning was performed 1-3 hr after intravenous injection of iomeprol-containing liposomes. RESULTS: After injection of iomeprol-containing liposomes at a dose of 70 mg of liposome-entrapped iodine per kilogram of body weight, the normal liver parenchyma showed a contrast enhancement, in Hounsfield units, of more than 60% over the control value before bolus. Liver tumors with no or few Kupffer cells were not enhanced and appeared as dark areas within the normal parenchyma. Tumors and pretumoral lesions devoid of Kupffer cells, as small as 3 mm in diameter, could be distinguished using this non-invasive method. CONCLUSION: CT liver scanning after injection of iomeprol-containing liposomes appears to be promising method for detecting liver tumors and focal liver lesions. PMID- 9419608 TI - Physicochemical properties, pharmacokinetics, and biodistribution of gadoteridol injection in rats and dogs. AB - RATIONALE AND OBJECTIVES: The physicochemical properties of gadoteridol, a macrocyclic nonionic gadolinium complex, were studied together with its pharmacokinetics and biodistribution in rats and dogs. METHODS: Studies in rats were conducted after single intravenous injections at 0.1 or 0.35 mmol/kg using 153Gd-labeled gadoteridol or with seven daily doses of 0.1 mmol/kg to examine the levels of residual gadolinium in organs. Nonradioactive biodistribution and excretion studies were performed in dogs following injection at 0.1 mmol/kg. RESULTS: After injection, the dose was rapidly cleared from rat blood and excreted such that more than 90% of the dose appeared in the urine within 4 hr of injection. At 7 and 14 days postinjection, only extremely low levels of gadolinium were observed in liver and bone; these levels were two to eight times lower than the levels reported after the injection of gadopentetate dimeglumine. CONCLUSION: The extracellular distribution and rapid urinary excretion of gadoteridol is in agreement with data obtained with other gadolinium-containing chelates used as intravascular magnetic resonance imaging contrast agents. Differences observed in the long-term retention of gadolinium between gadoteridol and gadopentetate dimeglumine were consistent with the reported greater in vivo resistance to transmetallation of gadolinium macrocycles compared with the linear gadolinium chelate molecules. PMID- 9419609 TI - A histogram-based algorithm for semiautomated three-dimensional craniofacial modeling. AB - RATIONALE AND OBJECTIVES: I conducted a study designed to facilitate thresholding and reduce volume averaging in three-dimensional (3D) computed tomography (CT) craniofacial modeling. METHODS: Three-dimensional CT reconstructions of orbits from two cadavers and seven clinical cases were generated from paired axial and coronal data. A histogram-based algorithm that was based on preliminary phantom and craniofacial specimen trials was applied to orbital data to identify volume averaged regions of thin bone to be used in conjunction with standard bone thresholds. Region-of-interest measurement of the orbital floors on the original two-dimensional slice data assessed algorithm performance. RESULTS: In five of the nine cases (55%), configuration of the superimposed histograms predicted regions of volume averaging. In only one case did such a region localize to the orbital floor. In the remaining four cases, the air-mucosa interface deep to the orbital floor was identified by the histogram method. CONCLUSION: Operator dependent editing remains superior to this histogram-based algorithm in reducing volume averaging in 3D craniofacial modeling. PMID- 9419610 TI - Videomanometry of the pharynx in dysphagic patients with a posterior cricopharyngeal indentation. AB - RATIONALE AND OBJECTIVES: Patients with a posterior indentation in the pharyngoesophageal segment (PES) are generally considered to have an abnormality of the cricopharyngeal muscle (CPM). In this study we determined the actual width of the PES and the pressure circumstances during swallowing within the pharynx and PES in such patients. METHODS: Simultaneous videofluoroscopy and solid state manometry, radiologic examination of the pharynx, PES, and cervical esophagus were performed in 16 dysphagic patients with a cricopharyngeal (CP) bar. In eight patients the indentation was 25-50%, and in eight it was more than 50% of the adjacent gullet. Sixteen dysphagic patients without a CP bar were used as control subjects. In each patient swallows of 10-ml barium bolus were recorded. RESULTS: Patients with CP bars had a significantly wider PES above (p = .0005) and below (p = .02) the CPM, whereas the diameter at the level of the CPM was smaller only in the patients with more than 50% indentation compared with the patients without a CP bar. The contraction pressure above the CP bar (i.e., at the level of the inferior pharyngeal constrictor) was significantly (p = .002) weaker in patients with a CP bar (131 +/- 16 mm Hg) than in those without a CP bar (222 +/- 20 mm Hg). CONCLUSION: Our findings in patients with a posterior CP bar suggest that the major abnormality is weak constrictors with outpouching of the gullet above and below. Only in patients with more than 50% indentation was there a slight narrowing at the level of the CP bar. The CPM showed no manometric abnormalities in terms of resting pressure, relaxation, and contraction pressure. Therefore, the CPM is likely to relax and distend normally during swallowing in patients with a CP bar. PMID- 9419611 TI - Two-dimensional intravenous coronary arteriography using above-K-edge monochromatic synchrotron X-ray. AB - RATIONALE AND OBJECTIVES: For intravenous (i.v.) coronary arteriography, a real time two-dimensional (2D) imaging system is being developed using above-K-edge monochromatic X-rays alone. The potential diagnostic value of this system was examined in the current study. METHODS: The angiographic system consisted of an asymmetric silicon (311) monocrystal, an image intensifier, and a charged-coupled device camera. It was constructed at the beam line of the Tristan Accumulation Ring. Monochromatic X-rays of 33.32 keV were used, and each image was recorded over 2 msec. RESULTS: Ventricular wall motion and the anatomy of the coronary arteries could be seen in dogs by sequential images obtained without subtraction. The left anterior descending coronary artery, left circumflex coronary artery, and right coronary artery and the branches of these vessels were observed. The parts of the coronary arteries overlapping the aorta and left ventricle were revealed somewhat during washout of the contrast material. CONCLUSION: For a 2D imaging system, monochromatic i.v. coronary angiography using an energy above the iodine K-edge might be able to image the coronary arteries without subtraction. However, the image quality needs to be improved by increasing the X-ray flux, decreasing background radiation scatter from the object, and decreasing contamination with 99-keV X-rays. PMID- 9419612 TI - Hemodynamic effects and image quality of low-osmolar ionic and nonionic contrast media during pulmonary angiography. AB - RATIONALE AND OBJECTIVES: We compared the hemodynamic responses to ionic and nonionic low-osmolar contrast media of patients who underwent pulmonary angiography. METHODS: Ninety-nine consecutive patients with suspected pulmonary emboli were randomly assigned to receive either 40 ml iohexol or 40 ml ioxaglate in 2 sec at 600 psi (0.17 kg/m2). Mean pulmonary arterial pressure, pulse rate, and blood pressure were recorded before, immediately after, and 2, 5, and 10 min following injection. Image quality was assessed by readers who were unaware of drug assignment. RESULTS: Pulmonary arterial pressure increased to a maximum at 2 min and was higher in patients with pulmonary emboli (p = .06). There were no significant differences between the two contrast media used. The systolic blood pressure and pulse rate in patients with pulmonary emboli increased significantly more in the ioxaglate group (ps = .03 and .04, respectively). Image quality was excellent in 90% of both groups. CONCLUSION: Both contrast agents are safe for pulmonary angiography and yield similar image quality. There appears to be a positive inotropic effect of ioxaglate. PMID- 9419613 TI - Evaluation of histopathologic changes in an animal model of mechanical corpus callosum impingement as seen in hydrocephalus. AB - RATIONALE AND OBJECTIVES: We evaluated histologic changes associated with chronic impingement of the corpus callosum. Similar callosal impingement has been postulated to be responsible for some of the symptoms in people who have hydrocephalus. METHODS: Eight rats with callosal impingement produced by surgical implantation of a blunt blade in the interhemispheric fissure and four control animals with no callosal impingement were evaluated by magnetic resonance (MR) imaging and by direct histologic evaluation after autopsy. The histologic evaluations occurred 1 month after surgery in half the animals and 6 months after surgery in the other half. RESULTS: MR imaging results showed that the implanted blade was in a good position in all animals. Histologically, the corpus callosum appeared normal 1 month after implantation of the impingement blade. Six months after surgery, the experimental group demonstrated decreased callosal thickness and a loss of axonal fibers in the corpus callosum both near and remote to the blade. CONCLUSION: Chronic impingement of the corpus callosum was associated with callosal thinning and by loss of callosal axons. Further research will be required to investigate the possible relation of these histologic findings to the clinical findings in normal-pressure hydrocephalus. PMID- 9419614 TI - Effects of a modified Q-switched Nd-YAG laser on human vascular tissue: an ex vivo study. AB - RATIONALE AND OBJECTIVES: Relatively disappointing results with continuous-wave lasers stimulated us to evaluate pulsed lasers for interventional radiology. In this article, we describe our efforts to assess the effects of this technology ex vivo. METHODS: We modified a Q-switched yttrium aluminum garnet (Nd-YAG) laser to emit pulses of 300 mJ maximum with a 20-Hz repetition rate, at 1064 nm, and with a duration that ranged from 300 ns to 2.3 microseconds. The lengthening of the pulse duration by a factor of 100 (compared with the conventional nanosecond Q switched Nd-YAG laser) and the ability to define it exactly were obtained by controlling the opening and closing of the Pockels cell electronically. Lengthening the pulse duration made it possible to reduce peak power while conserving the same total energy. In this way, high energy was transmitted through thin optical fibers. RESULTS: One hundred fifty millijoules with 2 microsecond pulses, 140 mJ with 1-microsecond pulses, and 100 mJ with 500-ns pulses were transmitted through a 300-micron silica-polymer fiber. The transmission coefficient was identical for the three pulse durations. Ex vivo irradiation experiments were performed on human atheromatous arteries in saline solution using a 300-micron diameter optical fiber. Craters were easily obtained. Their depth and width were related to maximum energy transmission and irradiation time. No carbonization occurred and no destruction of the optical fiber was observed. CONCLUSION: A modified Q-switched Nd-YAG laser can transmit high-energy pulses through thin optical fibers without damaging them and can destroy human atheroma in an ex vivo setting. PMID- 9419615 TI - Multireader, multicase receiver operating characteristic methodology: a bootstrap analysis. AB - RATIONALE AND OBJECTIVES: We evaluated by bootstrapping the conclusions obtained by the Dorfman-Berbaum-Metz (DBM) receiver operating characteristic (ROC) method and by the Toledano-Gatsonis (TG) method on a well-known data set. METHODS: We bootstrapped in two ways, resampled cases while holding readers fixed and resampled both cases and readers. RESULTS: When an analysis of variance of pseudovalues implies that reader variance and all random interactions with treatment are essentially zero, then case-resampling bootstrap and the DBM and TG methods should give the same results. Case-resampling bootstrap and the DBM and TG methods did give highly similar results for both individual readers and the averages over all readers. Both the case-resampling bootstrap and the reader-case resampling bootstrap gave smaller standard errors for group than for individual reader means, thereby providing evidence for a trade-off of readers and cases with regard to precision and power in this data set. CONCLUSION: Case-resampling bootstrap provides some justification for the DBM and TG methods. PMID- 9419616 TI - Characterizing the performance of diagnostic imaging systems. PMID- 9419617 TI - Painless paraspinal-gluteal mass. PMID- 9419618 TI - The fellowship trap. PMID- 9419619 TI - Evaluation of a breast biopsy phantom for freehand ultrasound guided biopsy of the liver. PMID- 9419620 TI - Computer-aided detection of clustered microcalcifications in digitized mammograms. AB - RATIONALE AND OBJECTIVES: We investigated a computer-aided detection (CAD) scheme for clustered microcalcifications in digitized mammograms. METHODS: A multistage CAD scheme was developed and tested. To increase sensitivity, the scheme uses a Gaussian band-pass filter and nonlinear threshold. A multistage local minimum searching routine and a multilayer topographic feature analysis are used to reduce the false-positive detection rate. One hundred ten digitized mammograms were used in this preliminary test, with 55 images containing one or two verified microcalcification clusters. RESULTS: The CAD scheme achieved 100% sensitivity and had an average false-positive detection rate of 0.18 per image. CONCLUSION: The CAD scheme performs as well as many published schemes and has some unique advantages to further improve detection sensitivity and specificity of future CAD schemes. PMID- 9419621 TI - Effect of arterial cannulation and contrast agents on blood coagulation. AB - RATIONALE AND OBJECTIVES: Nonionic contrast media have been considered by some to have thrombogenic properties. We prospectively assessed the effect of femoral artery catheterization and both nonionic and ionic contrast media on the coagulation parameters--fragment 1 + 2 (F1 + 2) and fibrinopeptide A (FpA)- during clinical angiography. METHODS: Seventeen patients undergoing aortography were included. Blood samples were obtained before and after arterial puncture and before and up to 30 min after contrast administration. RESULTS: An increase in FpA was observed after arterial puncture (range = 8.4 +/- 1.9 to 13.6 +/- 2.3 ng/ml, p < .004; data are written as mean +/- standard error of the mean). There was an observed increase in F1 + 2 after arterial puncture that was not statistically significant (2.0 +/- 0.4 to 2.3 +/- 0.4 nmol/l). No further increase was observed in either FpA or F1 + 2 levels after nonionic or ionic contrast media administration. CONCLUSION: The increased activity of the coagulation system during angiography is related to the arterial puncture, and nonionic and ionic contrast media have no thrombogenic potential in vivo. PMID- 9419622 TI - The meaning of a nonspecific abdominal gas pattern. AB - RATIONALE AND OBJECTIVES: The radiology report is the primary means of communication between the radiologist and the referring physician. A lack of precision in this report may adversely affect patient care. We examined how radiologists would define "nonspecific abdominal gas pattern" and how referring physicians would perceive the meaning. METHODS: A questionnaire was distributed to radiologists and referring physicians in Flint, Michigan. They were asked to categorize their definition or interpretation, respectively, of "nonspecific abdominal gas pattern" into "normal"; "either normal or abnormal"; or "abnormal, representing either mechanical obstruction or adynamic ileus." RESULTS: Thirty three radiologists responded, 23 (69.7%) of whom used the term. One hundred fifty seven referring physicians responded, 127 of whom assigned a specific definition to the term. Using a 2 x 3 chi-square test (df = 2), we found a statistically significant difference (p < .03) between the distribution of the meaning of the term between radiologists and their referring physicians. CONCLUSION: The term "nonspecific abdominal gas pattern" should be abandoned because it may signify a normal condition or a pathologic state. We found the definition to be dichotomous and asynchronous between radiologists and their referring physicians. PMID- 9419623 TI - Tissue ablation with radiofrequency using multiprobe arrays. AB - RATIONALE AND OBJECTIVES: We studied the feasibility of increasing the volume of tissue destroyed by radiofrequency tissue coagulation using multiprobe arrays and defined parameters that determine lesion size and shape. METHODS: Radiofrequency was applied to ex vivo calf liver using arrays of two to five 18-gauge probes for 6 min at 70-90 degrees C. Probe spacing (1-3 cm) and arrangement, as well as the method of radiofrequency application (simultaneous or sequential), were varied. The resulting areas of tissue coagulation were measured and compared. RESULTS: Uniform tissue necrosis was observed with simultaneous radiofrequency application for probes 1.5 cm or less apart. At 1.5 cm, arrays of three equidistant probes produced spheroid lesions approximately 3.0 +/- 0.2 cm in diameter. Arrays of four equidistant probes produced cuboid lesions of 3.2 +/- 0.1 cm per side. However, probes placed 2 cm or more apart produced independent lesions 1.4 cm in diameter, with incomplete necrosis between probes. In the trials using five-probe arrays, a central region 4mm in diameter showed no visible evidence of tissue necrosis. With each array, lesion size varied less than 3 mm in any direction. Greater necrosis was accomplished when radiofrequency was applied simultaneously rather than sequentially. CONCLUSION: Multiprobe radiofrequency arrays permit the destruction of more tissue in a single treatment session than is possible with multiple individual probes operating alone. Probes spaced 1.5 cm or less apart act synergistically, producing a total volume of coagulated tissue that is greater than when the individual probes are operated sequentially. PMID- 9419624 TI - Urine profiles and kidney histology after ionic and nonionic radiologic and magnetic resonance contrast media in rats with cisplatin nephropathy. AB - RATIONALE AND OBJECTIVES: The nephrotoxic drug cisplatin has been used successfully in treating some cancers. Patients with suspected carcinoma frequently undergo examinations with contrast media. We examined whether ionic and nonionic radiologic and magnetic resonance contrast media would have any effect on cisplatin nephropathy in rats. METHODS: Urine and serum profiles were monitored for 24 days after intravenous (i.v.) injections of saline, diatrizoate, iohexol, gadopentetate dimeglumine, and gadodiamide in high doses (4.59 mmol/kg body weight) in rats that received a weekly intraperitoneal (i.p.) injection of cisplatin (1 mg/kg) for 10 weeks. There were 10 rats in each group. Another 10 rats injected with both i.p. and i.v. saline served as control subjects. After euthanization, rats' kidneys were removed for examination by light microscopy and electron microscopy. RESULTS: Light and electron microscopy showed severe morphologic changes, including tubular dilatation, atrophy, and necrosis induced by cisplatin; however, the contrast media did not induce any additional morphologic changes. Gadopentetate dimeglumine, diatrizoate, and iohexol significantly increased (3-20 times) albuminuria compared with i.v. saline in cisplatin nephropathy, whereas gadodiamide did not. Albuminuria was highest after diatrizoate injection. All four contrast media caused an immediate and transient significant increase in the excretion of the brush border enzymes alkaline phosphatase and gamma-glutamyltransferase (125-500 times) and the cytoplasmatic enzymes alanine aminopeptidase and lactate dehydrogenase (16-100 times). Compared with saline, the ionic agents significantly increased the excretion of both glucose (two times) and sodium (three to five times), whereas the nonionic agents did not. CONCLUSION: High doses of radiologic and magnetic resonance contrast agents cause temporary dysfunction in rats with cisplatin nephropathy. Gadodiamide caused the least dysfunction and diatrizoate the most. PMID- 9419625 TI - Immediate and delayed tolerance of iohexol and ioxaglate in lower limb phlebography: a double-blind comparative study in humans. AB - RATIONALE AND OBJECTIVES: We compared the tolerance and efficacy of iohexol-300, a nonionic low-osmolar monomer, with those of ioxaglate-320, an ionic low-osmolar dimer, in lower limb phlebography. METHODS: One hundred twenty inpatients were randomly divided into two groups in this double-blind comparative study. Two hundred milliliters of contrast medium (100 ml per leg) was injected intravenously. The immediate tolerance was classified as discomfort (i.e., sensation of warmth, pain, coldness related to the injection) and adverse events occurring up to 1 hr after administration. Delayed tolerance was followed up to 8 days after the examination. The main parameter was immediate adverse events. Image quality was assessed by a radiologist using a visual analog scale. RESULTS: The number of immediate adverse events was significantly higher in the ioxaglate group (p < .02). The more frequent events were digestive disorders and skin rashes; 13 of these events were reported in the ioxaglate group, but none were reported in the iohexol group (p < .001). The other parameters were not significantly different in the two groups. CONCLUSION: We found a similar efficacy and a better tolerance of iohexol-300 than ioxaglate-320 in lower limb phlebography. PMID- 9419626 TI - Localization of metalloporphyrin-induced "specific" enhancement in experimental liver tumors: comparison of magnetic resonance imaging, microangiographic, and histologic findings. AB - RATIONALE AND OBJECTIVES: We investigated the tumor specificity of gadolinium mesoporphyrin (Gd-MP) and manganese tetraphenylporphyrin (Mn-TPP) as magnetic resonance (MR) imaging contrast agents. METHODS: Fifteen rats with multiple hepatocellular carcinomas and eight rats with implanted Novikoff hepatomas were given intravenous injections of either Gd-MP or Mn-TPP at 0.05 mmol/kg, which was compared with nonspecific gadopentetate dimeglumine (0.3 mmol/kg). T1-weighted spin-echo images were obtained before and up to 48 hr after injection and compared with corresponding microangiograms and histologic specimens. The relative enhancement of organs and tumors was plotted as a function of time. RESULTS: Initially, both metalloporphyrins behaved as nonspecific agents, similar to gadopentetate dimeglumine, and enhanced the tumor by perfusion and diffusion. However, metalloporphyrins, but not gadopentetate dimeglumine, caused a delayed (> or = 3 hr) enhancement in some compartments of certain lesions. The MR imaging microangiography-histology matching technique revealed that those compartments were actually nonviable components, including necrosis (n = 10), thrombosis (n = 7), and cystic secretion (n = 3), but not viable tumor tissue. CONCLUSION: Metalloporphyrins did not prove to be tumor specific. However, the observed affinity for nonviable tissue has elicited other potential applications for these agents. PMID- 9419628 TI - Two prototype blood-pool agents for contrast-enhanced magnetic resonance angiography of the portal vein in pigs. AB - RATIONALE AND OBJECTIVES: Macromolecular "blood-pool" agents such as polylysine gadopentetate dimeglumine or albumin-gadopentetate dimeglumine, which provide prolonged intravascular enhancement, were tested as magnetic resonance (MR) angiography contrast agents for the portal vein in pigs. METHODS: Phase-contrast MR angiography of the portal veins was performed on six pigs before and after intravenous administration of polylysine-gadopentetate dimeglumine (n = 3) or albumin-gadopentetate dimeglumine (n = 3). RESULTS: The contrast-to-noise ratio of the portal veins was improved by 74% and 52%, respectively, using polylysine gadopentetate dimeglumine or albumin-gadopentetate dimeglumine. More branches of the intrahepatic portal veins were visualized on postcontrast images. CONCLUSION: Blood-pool paramagnetic contrast agents improved the visualization of hepatic vasculature using phase-contrast MR angiography in our experimental model. PMID- 9419627 TI - Hepatic imaging with iodinated nanoparticles: a comparison with iohexol in rabbits. AB - RATIONALE AND OBJECTIVES: We evaluated the efficacy of a particulate computed tomography (CT) contrast agent in an animal model of focal liver disease. METHODS: Ethyl ester of diatrizoic acid (EEDA) is an iodinated (89 mg I/ml) nanoparticulate (200 nm) contrast agent intended for intravenous use that is currently undergoing preclinical testing in our laboratory. Focal liver abscesses were created in 11 New Zealand White rabbits. Iohexol and EEDA were administered to each animal on different days. CT scanning was performed at intervals following contrast agent administration. Liver and abscess enhancement were measured and compared. Dynamic imaging experiments in normal animals were also performed using both agents. RESULTS: EEDA resulted in significantly greater enhancement of the liver and liver-to-abscess contrast than did iohexol at all time points beyond 5 min at approximately 25% of the total iodine load. During dynamic imaging, liver and aortic enhancement were greater with EEDA than with iohexol, except during a 20- to 40-sec period immediately following contrast agent administration. CONCLUSION: EEDA is superior to iohexol for imaging liver abscesses. Our results suggest that liver-directed agents such as EEDA may prove to be more efficacious than currently available extracellular agents designed for liver CT scanning. PMID- 9419629 TI - Multireader receiver operating characteristic studies: a comparison of study designs. AB - RATIONALE AND OBJECTIVES: Traditionally, multireader receiver operating characteristic (ROC) studies have used a "paired-case, paired-reader" design. The statistical power of such a design for inferences about the relative accuracies of the tests was assessed and compared with alternative designs. METHODS: The noncentrality parameter of an F statistic was used to compute power as a function of the reader and patient sample sizes and the variability and correlation between readings. RESULTS: For a fixed-power and Type I error rate, the traditional design reduces the number of verified cases required. A hybrid design, in which each reader interprets a different sample of patients, reduces the number of readers, total readings, and reading required per reader. The drawback is a substantial increase in the number of verified cases. CONCLUSION: The ultimate choice of study design depends on the nature of the tests being compared, limiting resources, a priori knowledge of the magnitude of the correlations and variability and logistic complexity. PMID- 9419630 TI - Completeness and validity of a radiologic-epidemiologic database in Sweden. PMID- 9419631 TI - Radiology and the Internet. PMID- 9419632 TI - Abdominal mass in an 11-month-old girl. PMID- 9419633 TI - Child abuse: what one needs to know. PMID- 9419634 TI - Potential and limitations of magnetic resonance imaging for real-time monitoring of interstitial laser phototherapy. AB - RATIONALE AND OBJECTIVES: Magnetic resonance (MR) imaging has been suggested as a method to monitor interstitial laser phototherapy (ILP) in deep tissues. Unfortunately, a reliable relation between temperature and MR parameters has not yet been demonstrated. In this study, we examined whether such a relation exists and whether MR imaging can measure absolute temperature or temperature changes. METHODS: We evaluated, in the range of 21 degrees C to 80 degrees C, the temperature dependence of the MR imaging signal and T1 in samples of liver, water, CuSO4, and oil. Spin-echo and fast low-angle shot (FLASH) sequences were used. RESULTS: The MR imaging signal of liver, CuSO4, and water continuously decreased when the temperature was increased from 21 degrees C to 80 degrees C. By contrast, the MR imaging signal of the oil increased with increasing temperature up to 40-50 degrees C and then decreased at higher temperatures. We observed a reliable linear relation only between T1 and temperature in a range' of 30-60 degrees C for oil and CuSO4. CONCLUSION: MR imaging has the potential to measure thermal variations with an uncertainty of approximately +/- 10 degrees C. However, the use of MR imaging to monitor the real-time thermal effect induced in biologic tissues during laser irradiation requires further investigation before it can be applied clinically. PMID- 9419635 TI - Enhanced displays of medical images: evaluation of the effectiveness of color, motion, and contour for detecting and localizing liver lesions. AB - RATIONALE AND OBJECTIVES: Many perceptual studies have shown that the detection of large, low-contrast targets is better either in color or in contrast-reversing presentations than in standard gray scale. We determined the value of several new display techniques for viewing liver computed tomography (CT) scans. METHODS: Eight observers (four radiologists and four nonradiologists) viewed sets of 100 liver CT images (50 with lesions and 50 without) under five display conditions on a Macintosh computer: (1) color (equiluminant color contrast); (2) color luminance (combined luminance and chromatic contrast); (3) flicker (luminance contrast that reversed polarity at 2 Hz); (4) contour (shaded intensity mapping); and (5) control (conventional gray scale). Receiver operating characteristics (ROC) techniques were used for analysis. RESULTS: The measured ROC curve areas for the different viewing conditions were as follows: control = 0.77 +/- 0.01 (mean +/- standard error of the mean); color = 0.78 +/- 0.01; color-luminance = 0.82 +/- 0.01; flicker = 0.78 +/- 0.01; and contour = 0.76 +/- 0.01. The percentage of lesions correctly located ranged from 0.82 (color-luminance) to 0.75 (flicker). Performance under the color-luminance condition was significantly better than in the control condition (p = .01), whereas the other experimental conditions were not significantly different from the control condition (p > .21). CONCLUSION: The use of mixed color and luminance displays may have perceptual advantages for radiologists and can improve performance over that of gray-scale viewing. PMID- 9419636 TI - 19F chemical shift imaging technique to measure intracellular pO2 in vivo using perflubron. AB - RATIONALE AND OBJECTIVES: There is a linear relation between the T1 relaxation rate of fluorine-19 (19F) of perfluorochemicals (PFCs) and the partial pressure of the oxygen (pO2) dissolved in the PFC. A line scan technique was used to overcome the significant chemical shift and low signal-to-noise ratio (SNR) of in vivo 19F magnetic resonance imaging. This study was designed to determine whether the line scan technique could detect the effect of oxygen on 19F T1. In addition, its ability to detect changes in intracellular pO2 when the inspired gas was raised from 20% to 100% O2 also was investigated. METHODS: The T1 relaxation rate of samples of perflubron emulsion diluted from 3.5% to 70% w/v and equilibrated with N2-O2 gas mixtures (pO2 range = 10-450 mm Hg) was measured using the line scan technique. The gas and emulsion pO2 were measured with a blood gas analyzer. The liver T1 relaxation rate was measured in three rabbits given 5 ml/kg perflubron emulsion 4 and 8 days earlier as they breathed room air and then 100% O2. We used a prototype cylindrical coil double-tuned to hydrogen-1 (1H) and 19F and selected a line through the liver. The scanning parameters yielded a voxel size of 20 x 20 x 15.6 mm. Liver and blood samples were obtained postsacrifice for perflubron concentration measurement. RESULTS: A linear relation between the 19F T1 relaxation rate (1/T1) of the 3.5% w/v emulsion and dissolved pO2 was established with a slope of 0.0033 (sec-1/mm Hg) and a correlation coefficient of .991. As the PFC concentration increased by 1,900%, the slope increased by 21.2%. The 1/T1 for the liver was 0.182 +/- 0.004 sec-1 at baseline. It increased to 0.247 +/- 0.022 sec-1 when rabbits breathed 100% O2 (p = .023), which corresponded to an increase in intracellular pO2 of 19.7 mm Hg. The liver-to blood PFC concentration ratio was 500:1. CONCLUSION: In vitro measurements with the line scan technique replicated the established linear dependence of 1/T1 on pO2. In vivo measurements indicated a 20-mm Hg increase in intracellular pO2 of liver phagocytes when the inspired gas was changed from 20% to 100% O2. PMID- 9419637 TI - Tumor imaging with a macromolecular paramagnetic contrast agent: gadopentetate dimeglumine-polylysine. AB - RATIONALE AND OBJECTIVES: We evaluated magnetic resonance (MR) contrast enhancement of tumor tissue following injection of the macromolecular conjugate, gadopentetate dimeglumine-polylysine. METHODS: T1-weighted MR imaging scans were performed on female Fisher-344 rats with subcutaneously implanted mammary adenocarcinoma tumors. Following the baseline scan, gadopentetate dimeglumine polylysine or gadopentetate dimeglumine was injected at a dose of 0.1 mmol gadolinium per kilogram. RESULTS: Gadopentetate dimeglumine-polylysine injection resulted in a maximum enhancement of tumor contrast of 310 +/- 60% (n = 7). Tumor tissue remained enhanced and well defined for several days after gadopentetate dimeglumine-polylysine injection. Gadopentetate dimeglumine injection at the same dose resulted in a 70 +/- 25% (n = 4) maximal tumor enhancement and a corresponding 25 +/- 4% muscle enhancement. CONCLUSION: Gadopentetate dimeglumine polylysine provides higher, more sustained tumor contrast than does gadopentetate dimeglumine for the same dosage of gadolinium. PMID- 9419638 TI - Effect of a main renal artery stenosis on the downstream Doppler waveform in dogs. AB - RATIONALE AND OBJECTIVES: We evaluated the changes in the down-stream Doppler waveforms caused by a proximal stenosis in the main renal artery of dogs. METHODS: Renal parenchymal arterial waveforms downstream from mild (< 50%), moderate (50-75%), and severe (76-95%) stenoses were compared with nonstenotic baseline waveforms in five mongrel dogs. Waveform shapes were categorized as biphasic or monophasic. The percentage of biphasic and monophasic waveforms was determined for each stenosis. The acceleration index (AI) and the acceleration time (AT) were determined using traditional and modified calculations (AI' and AT'). Late systolic deceleration (DS), diastolic deceleration (DD), and the resistive index (RI) also were measured. RESULTS: AT, AI', and AT' demonstrated significant differences between the severe stenoses and nonstenotic baselines (p < .05); however, there was no difference between the mild and moderate stenoses versus baselines. The percentage distribution of monophasic and biphasic waveforms was highly correlated with the degree of stenosis. Monophasic waveforms increased on average from 22.5% of baseline waveforms to 76.5% of waveforms in the severe stenoses. Biphasic waveforms decreased on average from 69.9% of baseline waveforms to 18.7% of waveforms in the severe stenoses. CONCLUSION: Quantitative evaluation of the downstream waveform parameters (AI, AT, AI', AT', DS, DD, and RD in the dog kidney is not sufficiently accurate, but calculation of the percentage of the monophasic and biphasic waveforms present may be useful to predict a hemodynamically significant renal artery stenosis (> or = 50%). PMID- 9419639 TI - Radiofrequency tissue ablation in the rabbit lung: efficacy and complications. AB - RATIONALE AND OBJECTIVES: We assessed the feasibility and safety of performing percutaneous radiofrequency ablation of pulmonary tissue in rabbits. METHODS: Using an aseptic technique and computed tomography (CT) guidance, insulated 19 gauge aspiration biopsy needles were inserted into the right lower lobe of eight New Zealand White rabbits. Radiofrequency was applied via a coaxial electrode for 6 min at 90 degrees C. Probe-tip temperature, tissue impedance, and wattage were recorded at baseline and at 60-sec intervals throughout the procedure. CT scanning was used to assess tissue destruction and the presence or absence of pneumothorax immediately after the procedure and at 24 hr, 3 days, 10 days, 21 days, and 28 days. Three rabbits were sacrificed immediately, and the remaining rabbits were euthanized at 24 hr and at 3 days. 10 days, and 28 days (two rabbits). Gross and microscopic pathology were obtained and correlated with CT findings. RESULTS: The mean initial tissue impedance was 509 +/- 197 omega, marked changes in tissue impedance were found during the procedure (240-1380 omega). Rigid temperature control required continuous manual fine-tuning of generator output. Increased respiratory rate was noted in one rabbit during the first 30 sec of radiofrequency application. Homogeneous, ovoid opacities 8.4 +/- 2.4 mm in diameter and 1.4 +/- 0.1 cm in length were found by CT scanning immediately after the procedure. These opacities showed maximal consolidation at 3 days, corresponding to coagulative necrosis and a peripheral acute inflammatory reaction. At 10 days, peripheral hyperattenuation with central hypoattenuation (early fibrosis surrounding degenerating blood products) was seen. Minimal residual fibrosis, pleural scarring, or both were noted by 28 days, suggesting a rapid, near-total recovery from the procedure. Lesion sizes were within 2 mm of gross pathologic findings. Pneumothoraces were noted in three of the eight rabbits (37.5%). CONCLUSION: Radiofrequency tissue ablation was safely performed in pulmonary parenchyma via a percutaneous, transthoracic approach using a coaxial needle technique. Tissue response to thermal injury was predictable and easily monitored by CT scanning with excellent radiologic-pathologic correlation. PMID- 9419640 TI - Methodologic aspects of computed microtomography to monitor the development of osteoporosis in gastrectomized rats. AB - RATIONALE AND OBJECTIVES: We investigated the methodologic development of computed microtomography (CMT) for monitoring the development of osteoporosis in male Sprague-Dawley rats. METHODS: Eight rats were gastrectomized and eight rats were sham operated. Femurs, tibias, and tails were prepared, and CMT scans with spatial resolutions of 5-500 microns were made. Bone diameters, bone areas, and moments of inertia were determined from the CMT scans. Optimal slice position and the need for spatial resolution and energy optimization for future in vivo applications were investigated. RESULTS: Gastrectomy caused dramatic changes in the bone architecture of the tibia and the femur. The main features were vacuolization of the bone and reduced amounts of compact bone. Although the outer diameters of tubular bones (femur and tibia) were largely unaffected, their inner diameters were greatly increased following gastrectomy. Relative bone area and moment of inertia were greatly reduced. The optimal photon energy was 12 keV. CONCLUSION: It is possible to monitor gastrectomy-evoked changes in bone morphology at various sites in rats using CMT scanning. The changes are suggestive of osteoporosis. By optimizing the energy spectrum and spatial resolution, as well as choosing the proper slice position, it should be possible to keep absorbed doses low enough to avoid acute radiation injury in repeated in vivo measurements. PMID- 9419641 TI - Magnetization transfer in protein solutions at 0.1 T: dependence on concentration, molecular weight, and structure. AB - RATIONALE AND OBJECTIVES: We observed the magnetization transfer rates in a variety of protein solutions at 0.1-T magnetic field and compared our results with previous investigations at high magnetic fields (> 0.5 T). The effects of protein concentration, size, pH, denaturation, cross linking, and fiber formation were investigated. METHODS: We used the saturation transfer technique to determine the transfer of magnetization in gamma globulin, fibronectin, collagen, fibrinogen, and albumin solutions. RESULTS: The observed transfer rate increased with increasing concentration and size of the protein. Protein degradation decreased the transfer rate. Cross linking and fiber formation each increased the transfer rate, whereas buffer pH had no effect. CONCLUSION: Protein denaturation, aggregation, and fiber formation are important determinants of magnetization transfer in vitro. The size, concentration, and cross linking of the proteins contribute strongly to the transfer of magnetization at low fields, and the effect seems to be at least as important as at the higher fields. PMID- 9419642 TI - Phase-contrast imaging with synchrotron X-rays for detecting cancer lesions. AB - RATIONALE AND OBJECTIVES: We obtained image contrast in pathologic specimens without the use of contrast material by using phase-contrast imaging with synchrotron X-rays. METHODS: An experiment was performed at the three-pole superconducting vertical wiggler beam line BL-14B at the Photon Factory in Tsukuba, Japan. The X-ray phase-contrast imaging system consisted of a double crystal monochromator, an asymmetrically cut crystal monochromator, a triple Laue case X-ray interferometer, and film. The pathologic specimen was a sample from human liver that had metastatic carcinoma. RESULTS: The X-ray phase-contrast images of the pathologic specimen clearly depicted the cancerous lesion without the use of contrast enhancement, and the image showed good correlation with the photograph of the specimen. The X-ray absorption image did not differentiate between the normal liver tissue and the tumor. CONCLUSION: The results of this preliminary experiment reveal that for materials such as biologic specimens with a low atomic number, X-ray phase-contrast imaging better differentiates tissues than does the absorption contrast imaging commonly used in radiology. PMID- 9419643 TI - Pregnancy and maternity policies in radiology residencies: the 1993 survey of the American Association for Women Radiologists. PMID- 9419644 TI - The European Congress of Radiology. PMID- 9419645 TI - Night call in U.S. radiology residency programs. AB - RATIONALE AND OBJECTIVES: We acquired information about resident call in radiology programs throughout the United States to allow programs to compare themselves with others and to learn of possible alternate approaches to similar clinical and educational needs. METHODS: A 30-question survey was mailed to the program directors of all accredited U.S. radiology residency programs. A second mailing was sent to program directors who did not respond within 3 months. The survey addressed questions of the timing of call during residency training, the frequency of call, the nature of the call experience, and the relation to fellow and faculty call. Questions regarding available technical assistance, resident clinical activities postcall, faculty review of oncall studies, and other relevant issues were included. RESULTS: One hundred sixty-six of 206 (81%) of the program directors responded. The amount and type of call taken by radiology residents was highly variable in different programs. The mean number of in-house call days per month was 2.9 (SD = 2.6), 3.3 (SD = 2.2), 2.4 (SD = 1.9), and 1.3 (SD = 1.6) for first-, second-, third-, and fourth-year residents, respectively. The nature of technical and fellow/faculty assistance available to the resident was also variable. CONCLUSION: Night call in U.S. radiology residency programs is variable but tends to be concentrated in the second year of residency. Fourth year residents take less call than other residents, especially close to the time of the written and oral board examinations. Although program directors were satisfied with many aspects of their call systems, most indicated at least one major change they would like to make. PMID- 9419646 TI - Graduate medical education in radiology: a proposal for subspecialty training during residency. PMID- 9419647 TI - Relationships among the subjective quality, magnetic resonance spectra, and price of wine: a randomized trial. AB - RATIONALE AND OBJECTIVES: We sought to discriminate among wines on the basis of three techniques: physical properties such as smell, taste, and quality; market price; and chemical analysis using proton nuclear magnetic resonance (MR) spectroscopy. METHODS: A randomized, double-masked, controlled crossover wine tasting trial was conducted. Participants included seven men and seven women affiliated with an urban academic medical center, half of whom were physicians. The interventions consisted of eight red and eight white wines, including two respective, lower priced control wines. Each subject sampled all wines. Participants rated the overall quality of wine samples on a 5-point scale. The outcome measures were mean wine quality score, market price, and visual analysis of proton nuclear MR spectra. RESULTS: One subject dropped out. Three white wines (ps = .0245, .0275, and .0425) and two red wines (ps = .0072 and .0128) were rated significantly higher than their respective, lower priced control wines. The mean wine quality score was not significantly correlated with market price (white wine, rho = .371, p = .326; red wine, rho = -.072, p = .8492). Visual analysis of proton nuclear MR spectra from the highest scoring wines and their respective control wines revealed more similarities than differences. CONCLUSION: Quality assurance of wine may best be left to the discriminating palate rather than market price or visual analysis of proton nuclear MR spectra. PMID- 9419648 TI - On the perception of the left thoracic paraspinal line. PMID- 9419649 TI - Computer-aided diagnosis of breast cancer: artificial neural network approach for optimized merging of mammographic features. AB - RATIONALE AND OBJECTIVES: An artificial neural network (ANN) approach was developed for the computer-aided diagnosis of mammography using an optimally minimized number of input features. METHODS: A backpropagation ANN merged nine input features (age plus eight radiographic findings extracted by radiologists) to predict biopsy outcome as its output. The features were ranked, and more important ones were selected to produce an optimal subset of features. RESULTS: Given all nine features, the ANN performed with a receiver operator characteristic area under the curve (Az) of .95 +/- .01. Given only the four most important features, the ANN performed with an Az of .96 +/- .01. Although not significantly better than the ANN with all nine features, the ANN with the four optimized features was significantly better than expert radiologists' Az of .90 +/- .02 (p = .01). This four-feature ANN had a 95% sensitivity and an 81% specificity. For cases with calcifications, the radiologists' performance dropped to an Az of .85 +/- .04, whereas a specialized three-feature ANN performed significantly better with an Az of .95 +/- .02 (p = .02). CONCLUSION: Given only four input features, the ANN predicted biopsy outcome significantly better than did expert radiologists, who also had access to other radiographic and nonradiographic data. The reduced number of features would substantially decrease data entry efforts and potentially improve the ANN's general applicability. PMID- 9419650 TI - Tumor angiography using high-resolution, three-dimensional magnetic resonance imaging: comparison of gadopentetate dimeglumine and a macromolecular blood-pool contrast agent. AB - RATIONALE AND OBJECTIVES: We compared the peritumoral vascular definition in rats using either a paramagnetic extracellular or a macromolecular contrast medium in combination with high-resolution magnetic resonance (MR) imaging. METHODS: High resolution, three-dimensional spoiled gradient-refocused acquisition in a steady state (SPGR) images were acquired from tumor-bearing Fischer-344 rats before, immediately after, and again 40 min after administration of gadopentetate dimeglumine (0.1 mmol Gd/kg; n = 10) and albumin-(Gd-DTPA)30 (0.05 mmol Gd/kg; n = 5). Small peritumoral vessels were analyzed semiquantitatively on maximum intensity projection angiograms using a 4-point scoring system; quantitative analyses included signal-to-background ratios (SBRs) and signal-to-noise ratios. RESULTS: Gadopentetate dimeglumine caused a transient and low-scoring (0.2 +/- 0.1, SBR = 1.9 +/- 0.2) vessel definition but strong rim enhancement (score = 1.4 +/- 0.2). Albumin-(Gd-DTPA)30 produced persistent, high-quality angiograms (score = 2.6 +/- 0.2, SBR = 7.4 +/- 0.2) but minimal rim enhancement (score = 0.3 +/- 0.2). CONCLUSION: Albumin-(Gd-DTPA)30 combined with high-resolution MR imaging produces time-persistent, detailed angiographic definition of peritumoral vessels. Vascular maps obtained with gadopentetate dimeglumine enhancement are not time persistent or of equal quality. PMID- 9419651 TI - Quality of portal verification images using GLP7 film. AB - RATIONALE AND OBJECTIVES: To improve portal verification radiographs, we tested the application of GLP7 film. METHODS: The quality of the portal verification radiograph using the XV cassette-GLP7 film combination and the XL cassette-GLP7 film combination was investigated. The XV cassette-XV2 film combination and the SA cassette-GS screen-XTL film combination also were used for comparison. RESULTS: The characteristic curves showed that the relative speeds were 0.32 and 0.47 for the XV-GLP7 and XL-GLP7 combinations and that the average gradients were 1.93, 2.14, and 1.32 for the XV-GLP7, XL-GLP7, and XV-XV2 combinations, respectively. In the experiment using Burger's phantom, the smallest visible volumes were 11.8, 11.3, 28.7, and 19.6 mm3 for the XV-GLP7, XL-GLP7, XV-XV2, and SA-GS-XTL combinations, respectively. In the lower dosage treatment in the clinic, there were no marked differences between the GLP7 film and XV2 film. However, in the higher dosage treatment, the GLP7 film had a better quality than did the XV2 film. CONCLUSION: Portal verification radiographs using GLP7 film are of sufficient quality for clinical use. PMID- 9419652 TI - Multislice measurement of first-pass transit of gadobenate dimeglumine in normal and ischemic myocardium in dogs. AB - RATIONALE AND OBJECTIVES: We monitored the differences in the first passage of gadobenate dimeglumine through normal and ischemic myocardium with left anterior descending (LAD) coronary artery occlusion in dogs. METHODS: Dynamic multislice images of the heart were taken on a 1.5-T magnetic resonance (MR) imager. In six normal dogs, inversion recovery (IR)-prepared fast gradient-recalled echo (GRE) images were acquired at five doses of gadobenate dimeglumine (0.005-0.1 mmol/kg). First passage of the contrast medium through normal and acutely ischemic myocardium were monitored in seven dogs subjected to LAD coronary artery occlusion. RESULTS: IR-prepared GRE images showed a dose-dependent increase in the signal intensity (SI) of the myocardium. In dogs with LAD coronary artery occlusion, there was a significant increase in the SI of normal myocardium (p < .01) than in ischemic myocardium after injection of 0.025 mmol/kg gadobenate dimeglumine. CONCLUSION: The first-pass dynamics of gadobenate dimeglumine through normal and ischemic myocardium can be monitored with a multislice acquisition using a clinical MR imager and differentiated between normal and ischemic myocardium in dogs. PMID- 9419654 TI - Nonpulsatile arterial waveforms: observations during graded testicular torsion in rats. AB - RATIONALE AND OBJECTIVES: We tested whether testicular torsion could completely damp distal arterial pulsatility, resulting in venous-appearing arterial waveforms. METHODS: Progressively increasing testicular torsion was unilaterally produced in five rats. Doppler waveforms of the testicular artery distal to the torsion were obtained as soon as possible after each level of torsion until a complete absence of pulsatility was noted. RESULTS: One animal was not studied further after the first 180 degrees of torsion occluded flow. In three of the remaining four animals, the testicular artery resistive index (RI) at baseline (0.51, 0.58, 0.64) was within the range of the normal human intratesticular RI and decreased with increasing torsion, culminating in nonpulsatile, venous appearing waveforms at high degrees of torsion. CONCLUSIONS: Testicular torsion can completely damp arterial pulsatility, resulting in nonpulsatile, venous appearing arterial Doppler waveforms. PMID- 9419653 TI - Initial assessment of magnetoferritin biokinetics and proton relaxation enhancement in rats. AB - RATIONALE AND OBJECTIVES: We evaluated the biokinetics and proton relaxation enhancement of magnetoferritin, a recently developed class of superparamagnetic iron oxides, in rats. METHODS: "Equine" magnetoferritin was administered intravenously at 5 mg protein and 1.4 mg Fe/kg in nude rats carrying subcutaneous xenografted human small-cell lung carcinoma with and without preinjection of 100 mg/kg equine apoferritin. Blood clearance, in vivo biodistribution, and proton relaxation enhancement were assessed by variable field relaxometry, immunohistochemistry, and magnetic resonance (MR) imaging at 1.5 T. RESULTS: Magnetoferritin clearance from blood followed biexponential kinetics, with a short initial half-life of 1.4-1.7 min. A second, longer component lasted for several hours. Histochemical staining, MR imaging, and ex vivo relaxometry revealed rapid uptake of magnetoferritin in the liver, spleen, and lymph nodes. There was no difference in biodistribution after apoferritin preinjection. CONCLUSION: In the rat, equine magnetoferritin is rapidly sequestered by cells of the reticuloendothelial system, with no direct involvement of ferritin receptors. These properties may allow the use of magnetoferritin as an MR contrast agent for the liver and spleen. PMID- 9419655 TI - Contrast effect of blood on phase-contrast X-ray imaging. AB - RATIONALE AND OBJECTIVES: As a preliminary study for application of phase contrast X-ray imaging to medicine, we measured the X-ray refractive index of human blood and its influence on image contrast. METHODS: Using an X-ray interferometer, the X-ray phase shifts caused by human blood and its ingredients were measured from the interval of interference fringes. RESULTS: The X-ray phase shift caused by blood was 3% larger than that caused by serum. CONCLUSION: The large phase shift caused by blood is due to red blood cells, which contain hemoglobin molecules with iron atoms. It is possible that blood produces a structure in a phase-contrast X-ray image. PMID- 9419656 TI - Temporal bone volumetric image deblurring in spiral computed tomography scanning. AB - RATIONALE AND OBJECTIVES: We developed a method for volumetric image deblurring in spiral (helical) computed tomography (CT) scanning with a three-dimensional (3D) Gaussian point spread function (PSF) to improve the quality of temporal bone spiral CT images for assessing the position of cochlear implants electrodes. METHODS: A patient was scanned after cochlear implantation, and the temporal bone was reconstructed into a volume with 128 voxels per dimension, 0.1 mm per voxel side, and x 10 gray-scale expansion. The 3D PSF in spiral CT imaging was assumed to be Gaussian separable transversely and longitudinally. Standard deviations of the PSF were derived and subjectively adjusted. The image was then deconvolved using Wiener filtering and maximum-likelihood deconvolution methods. Image quality was assessed both visually and quantitatively using cross-sectional area at half of the maximum (CAHM) of the implanted array as the figure of merit. RESULTS: Substantial image deblurring was achieved via deconvolution. Subjectively, anatomic structures were more clearly shown. Deconvolution reduced the CAHM by approximately one third, on average. Three-dimensional deconvolution had better image quality than two-dimensional deconvolution. The maximum likelihood method produced superior image quality but took longer to process relative to Wiener filtering. CONCLUSION: Volumetric image deblurring is practical with a Gaussian PSF. The maximum-likelihood method is preferred if time permits. Deconvolution facilitates the study of fine details of the temporal bone and cochlear implant. PMID- 9419657 TI - A working cardiac valve phantom for radiographic assessment of prosthetic heart valves. AB - RATIONALE AND OBJECTIVES: A working valve phantom (WVP) that both exercises the valve occluder and simulates movements of the mitral annulus is described. It was designed to develop a method for radiographic detection of a single broken leg of the two-legged Bjork-Shiley convexo-concave (C/C) heart valve outlet strut. METHODS: The WVP consists of a pneumatically driven left ventricular assist device immersed in 22 cm of water. Left ventricular assist device annulus movements are generated by systolic turgor and diastolic relaxation of the aortic outflow graft within limits set by the holding fixture design. RESULTS: WVP images were comparable in attenuation, valve motion, and diagnostic sensitivity to clinical C/C valve images and were effective in assessing leaflet excursions in another valve model. Techniques developed in the WVP have proved successful in the clinical detection of C/C valves that have a single broken leg but that show normal function in all other tests. CONCLUSION: The WVP can be a useful tool for developing refined radiographic assessments of prosthetic heart valves. PMID- 9419658 TI - Design and evaluation of a flow phantom. AB - RATIONALE AND OBJECTIVES: We constructed a near-anatomically correct large-vessel phantom to perform repeatable flow dynamics research examinations by angiography, magnetic resonance (MR) angiography, and computed tomography (CT) angiography. METHODS: An internal carotid artery was constructed within a head phantom. The internal carotid artery branches into a middle and an anterior cerebral artery; the former trifurcates and ends in the superior sagittal sinus, and the latter ends in the inferior sagittal sinus. A transverse and sigmoid sinus drains the model. All four vessels connecting the arterial and venous vessels have variable flow-constricting ligatures placed around them. These ligatures are accessible on the skull surface. The skull cavity is filled with a silicone polymer that is isodense to brain on CT scans and isointense on most MR images. RESULTS: The flow in the phantom's vessels may be varied in a repeatable manner. Multiple scan sequences may be performed without the image degradation caused by patient motion. The homogeneity of the filler polymer allows visualization of flow related artifacts that may be hidden by complex human anatomy. CONCLUSION: Preliminary images of each modality show promise for use of the phantom in imaging research on large-vessel flow dynamics. PMID- 9419659 TI - Degeneracy and discrete receiver operating characteristic rating data. AB - RATIONALE AND OBJECTIVES: Observer performance studies sometimes use too few cases for estimating diagnostic accuracy from binormal receiver operating characteristic (ROC) curves. One important problem is degenerate data sets. We compared a new algorithm, RSCORE4, with the exact-solution approach to degeneracy in ROCFIT and with the Wilcoxon statistic. METHODS: Degenerate ROC solutions result from empty cells in the data matrix. We addressed this problem by adding a small constant to empty cells in a maximum-likelihood program, RSCORE4. When this method failed, the program branched to a pattern-search algorithm. We tested the program in a series of Monte Carlo studies. RESULTS: RSCORE4 converged to nondegenerate solutions in every case and gave results closer to population values than ROCFIT or Wilcoxon. ROCFIT converged to exact-fit degenerate solutions, those with zero or infinite parameter values, in more than 40% of the samples. The Wilcoxon statistic was biased. CONCLUSION: RSCORE4 seems to outperform other currently recommended methods for dealing with degeneracy. PMID- 9419660 TI - Functional brain imaging with a standard 1.5-T magnetic resonance imaging system. PMID- 9419661 TI - Academic radiology in crisis in the United Kingdom. PMID- 9419662 TI - Synchronous primary brain tumors. PMID- 9419663 TI - Balancing life. PMID- 9419664 TI - The Academy of Radiology Research. PMID- 9419665 TI - Comparison of color Doppler sonography and radionuclide imaging in different degrees of torsion in rabbit testes. AB - RATIONALE AND OBJECTIVES: We compared color Doppler sonography and radionuclide imaging in an animal model of variable torsion of the testes. METHODS: The testes of 19 rabbits with unilateral 0 degree (nontorsion), 180 degrees, 360 degrees, or 540 degrees torsion and contralateral nontorsion were evaluated by sonography serially over 24 hr. Color Doppler sonography and radionuclide imaging at 24 hr were compared and correlated with pathology in a subset of testes. RESULTS: Nontorsion (n = 16 testes) and 540 degrees torsion (n = 7 testes) were always correctly diagnosed at 24 hr by color Doppler sonography and radionuclide imaging. Torsion at 180 degrees (n = 2) was indistinguishable from nontorsion. With 360 degrees torsion (n = 6 testes), four testes had reduced or absent flow with color Doppler sonography, whereas only one testis was abnormal with radionuclide imaging. CONCLUSION: Nontorsion and extreme torsion of rabbit testes are well documented by radionuclide imaging and color Doppler sonography. Torsion at 360 degrees can result in variable flow alterations that are better detected by color Doppler sonography than by radionuclide imaging. PMID- 9419667 TI - Computerized detection of masses in digitized mammograms using single-image segmentation and a multilayer topographic feature analysis. AB - RATIONALE AND OBJECTIVES: We developed and evaluated a computer-aided detection (CAD) scheme for masses in digitized mammograms. METHODS: A multistep CAD scheme was developed and tested. The method uses a technique of single-image segmentation with Gaussian bandpass filtering to yield a high sensitivity for mass detection. A rule-based multilayer topographic feature analysis method is then used to classify suspected regions. A set of 260 cases, including 162 verified masses, was divided into two subsets; one set was used to set the rule based classification and one was used to test the performance of the scheme. RESULTS: In a preliminary clinical study, the implemented detection scheme yielded 98% sensitivity with a false-positive detection rate of less than one false-positive region per image. CONCLUSION: Single-image segmentation methods seem to have high sensitivity in selecting true-positive mass regions in the first stage of a CAD scheme. A multilayer topographic image feature analysis method in the second stage of a CAD scheme has the potential to significantly reduce the false-positive detection rate. PMID- 9419666 TI - Computed tomography evaluation of regional increases in microvascular permeability after reperfusion of locally ischemic myocardium in intact pigs. AB - RATIONALE AND OBJECTIVES: I evaluated how well fast computed tomography (CT) scanning could quantify altered myocardial microvascular function resulting from transient ischemia and subsequent reperfusion. METHODS: A major epicardial coronary artery of anesthetized pigs was occluded for either 0, 15, 30, 60, or 120 min. Fast X-ray CT scan sequences were performed before and during occlusion and 30, 60, and 90 min after onset of subsequent reperfusion. Regional myocardial perfusion, intramyocardial blood volume, and myocardial microcirculatory permeability-surface area product were estimated from the CT scan sequences. RESULTS: With the increasing duration of ischemia, the permeability-surface area product remained unchanged during reperfusion (p > .05), but an index of myocardial microvascular permeability increased with the progressive duration of ischemia and of reperfusion (p < .05). CONCLUSION: These CT-based findings are consistent with published, more invasive demonstrations that capillary permeability is increased during postischemic reperfusion in proportion to the duration of the preceding ischemia. PMID- 9419668 TI - Liver contrast enhancement in primates using iopromide liposomes. AB - RATIONALE AND OBJECTIVES: We studied the feasibility of using iodinated liposomes as computed tomography (CT) liver contrast agents in nonhuman primates. METHODS: Iopromide-containing liposomes were investigated as reticuloendothelial (RES) contrast agents for CT scanning of the liver in normal adult baboons. For intravenous (i.v.) injection, liposomes were resuspended in mannitol solution, filtered under sterile conditions, and injected i.v. at doses of 200 and 400 mg l/kg to each of five anesthetized adult baboons. RESULTS: Animals tolerated the injections without measurable electrocardiographic changes and recovered uneventfully from anesthesia. Sequential CT scans of the baboons' upper abdomen acquired up to 60 min postinjection showed persistent enhancement of the liver 10 60 min after injection. Maximum enhancement levels were 36 and 61 delta Hounsfield units (delta H) after the 200- and 400-mg/kg doses, respectively. The mean time to plateau enhancement was 20 min with the 200-mg/kg dose and 10 min with the 400-mg/kg dose. The greatest splenic enhancements were 181 and 301 delta H after the 200- and 400-mg/kg doses, respectively. CONCLUSION: Iopromide liposomes are effective as RES contrast agents in primates. PMID- 9419669 TI - Additive hemodynamic and electrophysiologic effects of repeated intracoronary contrast media injections in dogs with heart failure. AB - RATIONALE AND OBJECTIVES: We investigated the cardiac effects of single and repeated contrast media injections in dogs with heart failure and compared the effects of iohexol with iohexol supplemented with electrolytes (30 mmol/l NaCl, 0.15 mmol/l CaCl2, 0.9 mmol/l KCl, and 0.1 mmol/l MgCl2; iohexol + electrolytes [IPE]). Although it has a higher osmolality than iohexol, IPE appears to be safer when injected through a wedged catheter. METHODS: Acute ischemic heart failure was induced by injections of small plastic microspheres into the left coronary artery of 16 anesthetized dogs. Iohexol, IPE, and Ringer acetate were injected into the left coronary artery either as a 5-ml single injection or repeatedly five times, once every 10th second. RESULTS: Single injections of iohexol and IPE induced small hemodynamic and electrophysiologic effects. However, repeated injections of iohexol and IPE increased the maximum rate of isovolumetric contraction by 46% and 36%, reduced heart rate by 8% and 7%, and lengthened QTc (the Q-T interval corrected for heart rate) time by 44 and 39 msec, respectively. No statistically significant differences were found in a comparison of IPE and iohexol. CONCLUSION: During heart failure, repeated injections of iohexol and IPE induced similar additive hemodynamic and electrophysiologic effects without inducing arrhythmias or serious hemodynamic changes. PMID- 9419670 TI - Bronchial arterial response to contrast medium. AB - RATIONALE AND OBJECTIVES: Contrast agents have been shown to produce vasodilatory responses in several vascular beds. To our knowledge, however, their effect on the bronchial vasculature has not been examined. Clinically, contrast-induced bronchial vasodilation could potentially exacerbate life-threatening pulmonary hemorrhage during bronchial angiography for hemoptysis. In the current study, we systematically measured the bronchial vasodilatory response to diatrizoate meglumine 66% and diatrizoate sodium 10% (MD-76) and examined whether vasodilation would be mediated by nitric oxide (NO). METHODS: We measured bronchial blood flow in seven anesthetized, ventilated, open-chested sheep using an ultrasonic flow probe placed around the bronchial artery. Bronchial blood flow was recorded before and after injection of 2 ml MD-76 into the bronchial artery. The protocol was repeated after 20 min infusion of N omega-nitro-L-arginine (L NA; 10(-2) mol/l), an NO-synthase inhibitor, into the bronchial artery. RESULTS: There was a 45 +/- 8 ml/min increase (p < .01) in bronchial blood flow after injection of MD-76, which was reduced to 20 +/- 6 ml/min (p < .01) after infusion of L-NA. CONCLUSION: Bronchial arterial injection of MD-76 results in a consistent increase in bronchial blood flow that is mediated partly by NO. PMID- 9419671 TI - Indirect computed tomography lymphography using iodinated nanoparticles: time and dose response in normal canine lymph nodes. AB - RATIONALE AND OBJECTIVES: We evaluated the effect of time and dose on lymph node iodine uptake after subcutaneous or submucosal administration of iodinated nanoparticles used for computed tomography lymphography. METHODS: We injected 0.1 6 ml of a 15% wt/vol iodinated nanoparticle suspension into the distal extremities subcutaneously (n = 5) or into the buccal submucosa (n = 7) of normal dogs. Precontrast and 4, 12, 24, and 48 hr after contrast administration, CT scans of opacified lymph nodes were obtained. Iodine concentration, node volume, and total iodine uptake were estimated for each node. RESULTS: All estimated parameters increased between 4 and 12 hr postcontrast (p < .05), with no significant increase thereafter. At 24 hr postcontrast, iodine concentration ranged from 0.01 to 16.1 mg/ml (47-568 Hounsfield units). The average iodine concentration and total iodine uptake increased with contrast dose (p < .05) for all lymph node groups evaluated. Node opacification also revealed internal architectural detail. CONCLUSION: Subcutaneous and submucosal injections of iodinated nanoparticles result in a dose-dependent iodine uptake in targeted lymph nodes. In addition, architectural detail within opacified nodes can be visualized. PMID- 9419672 TI - Magnetic resonance imaging of the hepatobiliary system: intestinal absorption studies of manganese mesoporphyrin. AB - RATIONALE AND OBJECTIVES: We studied the intestinal absorption of manganese mesoporphyrin (Mn-mesoporphyrin), a potential oral hepatobiliary contrast agent. METHODS: Mn-mesoporphyrin was complexed with monoolein and taurocholate (mixed micelles). Portal venous delivery and biliary excretion were measured after intestinal administration in rats and rabbits, and the mechanism of intestinal transport was studied in a combined lymph-bile fistula model in rats. T1-weighted magnetic resonance (MR) images of the liver were obtained in rats and domestic pigs before and after gastric administration of Mn-mesoporphyrin in mixed micelles. RESULTS: A 2.2-fold increase of portal venous Mn concentration was found 90 min after intestinal administration of the complex. None was found in the lymph collected from the thoracic duct, indicating a transcellular transport mechanism through the intestinal mucosa with portal venous delivery. Mn mesoporphyrin levels in bile peaked between 240 and 270 min after administration (200-fold increase). The greatest liver enhancement (20-90%) was measured 360 min after administration. CONCLUSION: The feasibility of intestinal delivery of Mn mesoporphyrin, a lipophilic hepatobiliary contrast agent was demonstrated. PMID- 9419673 TI - An approach to radiologic quality assurance: analysis of medicolegal claims involving radiologic contrast-media-induced injury. PMID- 9419674 TI - Hiring preferences of diagnostic radiology groups: results of a 1994 survey. AB - RATIONALE AND OBJECTIVES: We surveyed diagnostic radiology group preferences and considerations in hiring radiologists and compared these findings with those of a survey performed in 1990. We sought to identify changes in hiring practices that might have occurred because of socioeconomic changes. We also sought to identify features of job candidates that make them more attractive to hiring groups. METHODS: One hundred surveys were mailed to a stratified random sample of diagnostic radiology groups identified by the American College of Radiology. We solicited information on the importance of various attributes and the level of experience of a candidate, the fellowship training considered most desirable, and the effect of changes in the health care socioeconomic environment. The responses were weighted by group size and geographic location to estimate what results might have been obtained if we had surveyed all groups in the United States. RESULTS: Seventy-five groups returned the survey. The two most important factors in choosing a candidate were motivation and radiologic knowledge. The fellowships that groups that were hiring considered to be the most desirable were body imaging, neuroradiology, and angiography/interventional radiology. Groups overwhelmingly preferred recent training over long experience. CONCLUSION: Fellowship training increases a candidate's marketability, but the two factors that hiring groups consider the most important are motivation and radiologic knowledge. PMID- 9419675 TI - The changing face of academic radiology: can it survive managed care? PMID- 9419676 TI - Learning radiology from interactive videodiscs: bar-code book versus computer assisted instruction. AB - RATIONALE AND OBJECTIVES: We compared an interactive videodisc with bar-code book with an interactive videodisc with computer-assisted instruction for learning radiology to determine whether there would be differences in instructional effectiveness, instruction time, or subjective preference. METHODS: Two different videodisc modules were created. Each was presented in two formats with identical content: bar-code book and computer-assisted instruction. In a controlled crossover experimental design, 48 fourth-year medical students were assigned one bar-code book module and one computer module. Pre- and posttests were administered. RESULTS: Mean scores improved from pretest to posttest after students used the modules in either format (p < .01). There were no significant differences between students who used the bar-code book and those who used the computer in pretest scores, posttest scores, gains in score from pretest to posttest, or instruction time for either module. Subjectively, 74% of the students preferred computer-assisted instruction, 20% preferred bar-code book, and 7% preferred neither. CONCLUSION: Although bar-code book and computer versions of an interactive videodisc can be educationally equivalent, most students preferred the computer. When videodisc is being integrated into the curriculum, the choice between bar-code book and computer-assisted instruction can be made on the basis of noneducational factors such as cost and availability. PMID- 9419677 TI - Chairperson's rounds for radiology residents. PMID- 9419678 TI - Lateral neck mass in a child. PMID- 9419679 TI - The radiology resident in difficulty. PMID- 9419680 TI - 1995 AUR Memorial Award. Gamma knife irradiation-induced changes in the normal rat brain studied with 1H magnetic resonance spectroscopy and imaging. AB - RATIONALE AND OBJECTIVES: The pathogenesis of brain injury following radiosurgery is poorly understood. To better elucidate the relationship between blood-brain barrier disruption and metabolic derangements, we used magnetic resonance (MR) imaging and 1H MR spectroscopy to detect early changes from focused single fraction, high-dose irradiation injury in rat brains. METHODS: Using the Leksell gamma knife, we irradiated the frontoparietal cortex of 11 male Wistar rats with a single dose of 120 Gy. Four weeks later, we sequentially performed water suppressed 1H MR spectroscopy and gadopentetate dimeglumine-enhanced T1-weighted MR imaging. Metabolic maps were created of n-acetylaspartate (NAA), creatine and choline (Cr/Cho), and lactate from the MR spectroscopy data set. Detection of irradiation injury among the tested modalities was assessed by receiver operating characteristic analysis and by quantitative signal intensity changes. Pathologic confirmation of irradiation damage was obtained in all rats. RESULTS: Gadopentetate dimeglumine-enhanced T1-weighted MR imaging was the only imaging modality that detected statistically significant signal intensity changes (p < .05). No reproducible changes in the metabolites of interest could be detected by 1H MR spectroscopy. CONCLUSION: In our animal model, blood-brain barrier disruption was a reproducible, integral finding of single-fraction, high-dose irradiation injury. No reproducible metabolic derangements of ischemia or necrosis were detected by 1H MR spectroscopy, possibly because of dose-latency effects or sensitivity issues. PMID- 9419681 TI - 1995 Joseph E. Whitley, MD, Award. A World Wide Web gateway to the radiologic learning file. AB - RATIONALE AND OBJECTIVES: Computer networks in general, and the Internet specifically, are changing the way information is manipulated in the world at large and in radiology. The goal of this project was to develop a computer system in which images from the Radiologic Learning File, available previously only via a single-user laser disc, are made available over a generic, high-availability computer network to many potential users simultaneously. METHODS: Using a networked workstation in our laboratory and freely available distributed hypertext software, we established a World Wide Web (WWW) information server for radiology. Images from the Radiologic Learning File are requested through the WWW client software, digitized from a single laser disc containing the entire teaching file and then transmitted over the network to the client. The text accompanying each image is incorporated into the transmitted document. RESULTS: The Radiologic Learning File is now on-line, and requests to view the cases result in the delivery of the text and images. Image digitization via a frame grabber takes 1/30th of a second. Conversion of the image to a standard computer graphic format takes 45-60 sec. Text and image transmission speed on a local area network varies between 200 and 400 kilobytes (KB) per second depending on the network load. CONCLUSION: We have made images from a laser disc of the Radiologic Learning File available through an Internet-based hypertext server. The images previously available through a single-user system located in a remote section of our department are now ubiquitously available throughout our department via the department's computer network. We have thus converted a single-user, limited functionality system into a multiuser, widely available resource. PMID- 9419682 TI - Computerized detection of masses from digitized mammograms: comparison of single image segmentation and bilateral-image subtraction. AB - RATIONALE AND OBJECTIVES: Two methods--single-image segmentation and bilateral image subtraction--have been used commonly as the first stage in computer-aided detection (CAD) schemes to detect masses on digitized mammograms. In the current study, we investigated and compared the advantages and disadvantages of the two methods in achieving a high sensitivity for mass detection. METHODS: Two CAD schemes were tested. One used Gaussian filtering based on single-image segmentation, and the other used bilateral-image subtraction based on left-right image pairs to identify suspicious mass regions. A clinical database that contained 152 verified mass cases was used to compare the two approaches. RESULTS: The single-image segmentation method yielded 100% sensitivity and had a somewhat higher number of initial suspicious regions. The bilateral-image subtraction method missed several true-positive regions at the initial phase. Each approach achieved more than 90% sensitivity at a false-positive rate of approximately 0.8 per image. CONCLUSION: Optimal initial image segmentation schemes may depend on the complete detection and classification method used. Single-image segmentation methods may perform comparably with bilateral-image segmentation schemes, and these techniques appear to be more versatile and easily adaptable to future clinical CAD applications. PMID- 9419683 TI - Correlation of total-body bone mineral content determined by dual-energy X-ray absorptiometry with bone mineral density determined by peripheral quantitative computed tomography. AB - RATIONALE AND OBJECTIVES: We sought to determine the value of peripheral quantitative computed tomography (pQCT) in measuring bone mineral density. METHODS: In 50 healthy, eugonodal premenopausal women, we correlated measurements of total bone mineral content (BMCTB), made with dual-energy X-ray absorptiometry (DXA), and bone mineral density, determined by pQCT. RESULTS: The partial correlations, adjusted for weight and age, between BMCTB and cortical bone density, total bone density, and trabecular bone density were .71 (p < .0001), .63 (p < .0001), and .32 (p < .05), respectively. CONCLUSION: These results and the advantages of pQCT--providing precise bone density determinations for trabecular and compact bone separately, having a high spatial resolution that allows a "compartmental" analysis of bone structure, having a low coefficient of variation, and having a minimal radiation dose (< 5 mrem)--confirm the adequacy of using this method for bone mass studies. PMID- 9419684 TI - Patient compliance in mobile screening mammography. AB - RATIONALE AND OBJECTIVES: We assessed the follow-up behavior of women who had abnormal results of screening mammograms taken on a mobile van. METHODS: A retrospective cohort study was conducted between 1988 and 1991 of all women served by a mobile mammography van in rural North Carolina. Compliance with radiologist recommendations for follow-up was assessed through review of patient records and mail surveys of patients with incomplete records. RESULTS: Compliance was 44% for negative or benign mammograms, 57% for indeterminate mammograms, and 62% for probably malignant or malignant mammograms. Women who had a previous mammogram or had a malignant finding were more likely to comply with follow-up recommendations (p < .0001) than women with normal or benign results and no history of mammography. Compliers and noncompliers did not differ with respect to family history of breast cancer or personal history of breast discomfort. CONCLUSION: Compliance with recommendations in this setting was lower than expected. This may be because rural women using mobile van mammography have limited access to the resources needed for appropriate follow-up. Further research is needed to examine explanations for poor compliance in this setting. PMID- 9419685 TI - Corpus cavernosum as an emergency vascular access in dogs. AB - RATIONALE AND OBJECTIVES: We investigated the feasibility of using the corpus cavernosum as an emergency vascular access in dogs with severe hypovolemia. METHODS: Five male mongrel dogs were brought to hypovolemic shock by withdrawal of blood through an internal jugular venous line. Using the corpus cavernosum as a venous access, normal saline was injected through a 23-gauge needle at the highest possible rate. Cavernosography was performed under fluoroscopy to verify the accurate position of the needle tip. Blood volumes were measured using 125I labeled human serum albumin. RESULTS: Before the dogs were bled, their mean systolic blood pressure was 121.4 (mean) +/- 5.2 (standard error) mm Hg and their mean blood volume was 1,835.2 +/- 139.7 ml. The mean volume of blood removed from the dogs was 710.0 +/- 67.8 ml. The mean systolic pressure during shock was 40.8 +/- 2.2 mm Hg. After fluid resuscitation, the mean systolic pressure recovered to 114.6 +/- 4.6 mm Hg and their mean blood volume increased to 1,763.2 +/- 112.7 ml. The mean rate of saline infusion into the corpus cavernosum was 50.2 +/- 0.7 ml/min. CONCLUSION: The results of this study demonstrate the feasibility of using the corpus cavernosum as an alternative route for fluid resuscitation in severely hypovolemic dogs. PMID- 9419686 TI - Polyurethane-coated, self-expandable biliary stent: an experimental study. AB - RATIONALE AND OBJECTIVES: We describe a self-expanding metallic biliary Gianturco Rosch stent coated with polymeric material. The coating was designed to prevent the growth of neoplastic and reactive tissue within the biliary ducts. METHODS: The stents were coated with a solvent-casting technique, which consists of dissolving polyurethane (polyether urethane or polycarbonate urethane) pellets in a solvent (dimethylacetamide), dipping the stent in the solution, and completely evaporating the solvent. In vitro mechanical characterization of the stent was performed to determine the adhesion of the coating to the metallic cage, the best introducer caliber for implantation of the device, and the relationship between the stent's diameter and radial stress. RESULTS: Reports in the literature on the biostability of polycarbonate urethane compared with polyether urethane prompted us to use the former material to coat the stents. The solvent technique gives a smooth internal surface of the stent wall, leaving in relief the coated structure of the stent on the external surface. The functional tests demonstrated that the coating did not compromise the original characteristics of the stent in terms of self-expandability, axial flexibility, and increased radial rigidity of the device. CONCLUSION: Functional tests verified coating stability and device handling, which are the first steps toward in vivo experimentation. PMID- 9419687 TI - Contrast-medium-induced ventricular fibrillation: arrhythmogenic mechanisms and the role of antiarrhythmic drugs in dogs. AB - RATIONALE AND OBJECTIVES: Small electrolyte additions to a nonionic contrast medium reduce the risk of ventricular fibrillation (VF) during wedged catheter injection of a contrast medium. The current study was designed to further investigate contrast-medium-induced VF by studying the effect of pretreatment with different antiarrhythmic drugs. METHODS: During a simulated wedged catheter situation, iohexol was injected into the anterior descending branch of the left coronary artery in five open-chest, anesthetized dogs pretreated with lidocaine, propranolol, amiodarone, almokalant, or verapamil. RESULTS: Wedging the catheter for 60 sec did not induce VF. However, all 15 wedged catheter injections with iohexol induced VF within 28 sec (19 +/- 1 [mean +/- standard error of the mean]) despite pretreatment with antiarrhythmic drugs. Prior to VF, conduction was slowed and monophasic action potential duration lengthened in the contrast-medium perfused myocardium, although no significant changes occurred in the control area. CONCLUSION: The combination of catheter wedging and long-lasting contrast medium injection has a high risk of causing VF. Although adding a small amount of electrolytes to nonionic contrast media can reduce the risk of VF, antiarrhythmic drug therapy may not have a protective effect. PMID- 9419688 TI - 1995 A3CR2 Malcolm D. Jones and Hartman Centennial Lecture. Postgraduate education in radiology: a historical review and future perspective. PMID- 9419690 TI - Top 10 ways for academic radiology departments to survive hard financial times. PMID- 9419689 TI - 1995 AUR Hartman Centennial Lecture. Academic radiology: time for action. AB - To summarize, the 10 actions I believe we should take are as follows: 1. Protect our patient base by institutional involvement and selected departmental outreach programs. 2. Reorganize our faculties and gain their support to meet the changes that will occur as a result of health care reengineering. 3. Restructure our residency and fellowship programs to adapt positively to the needs of a new delivery system. 4. Take a stand on resident/fellow training, accreditation issues, and program length and composition. 5. Develop a national program to continue to attract the best medical students into radiology. 6. Get the information needed to provide the best estimate of work force requirements and work toward achieving the proper balance between supply and demand. 7. Support subspecialization in our field. Quality eventually will be an issue. 8. Support research training for faculty and make research important. 9. Continue to present our field as an exciting place to be, which it is. 10. Support the AUR, the SCARD, and the APDR as the collective voice for academic radiology. Finally, I would like to challenge the AUR, the SCARD, and the APDR to unite to become a strong force in academic radiology. Academic radiology now has no singular voice. Radiologists in private practice have the ACR, neuroradiologists have the ASNR, vascular-interventional radiologists have the SCVIR, nuclear medicine radiologists have the Society of Nuclear Medicine, ultrasonographers have the American Institute of Ultrasound in Medicine, and I can name many other important special interest groups within our field. No doubt, all these organizations share many of our common concerns and interests, but having an organization interested solely in the continued health of academic radiology is vital to our future and, because of the reengineering of the health care system, more important than ever. Academic radiology is in the unique position of being radiology's only supplier of human resources and research. We must make our voice heard on staff issues, training program changes, accreditation, certification, financing of graduate education, support for biomedical research, and all other matters important to our academic programs. Although we will be drawn into the delivery of managed care, we cannot afford to ignore our other two principal missions: education and research. If we do, all of radiology will suffer the consequences in years to come. So let's take up the challenge and make our three organizations a steady, consistent, rational but firm voice in the support of academic radiology. I believe that the future of radiology is bright, but we need to make it happen. Now is the time for action. Thank you for the invitation to present the Glen W. Hartman lecture. It is truly a great honor. PMID- 9419691 TI - Minimizing the impact of health care reform on resident education. PMID- 9419692 TI - Resident education in the health care reform environment: the role of organized radiology. PMID- 9419693 TI - Decreasing the number of radiology residents: the impact on radiology departments and resident education. PMID- 9419694 TI - The impact of health care reform on research during residency training. PMID- 9419695 TI - Deciding where to make cuts in resident education: a resident's perspective. PMID- 9419696 TI - Organization of a radiology teaching program in a changing health care environment. PMID- 9419697 TI - Managed care residencies. PMID- 9419698 TI - 1995 AUR Gold Medal Awards. David Gordon Bragg, MD. PMID- 9419699 TI - 1995 AUR Gold Medal Awards. George Leopold, MD. PMID- 9419700 TI - Random-effects models in the receiver operating characteristic curve-based assessment of the effectiveness of diagnostic imaging technology: concepts, approaches, and issues. PMID- 9419701 TI - Random-effects models for diagnostic accuracy data. PMID- 9419702 TI - Multireader, multimodality receiver operating characteristic curve studies: hypothesis testing and sample size estimation using an analysis of variance approach with dependent observations. PMID- 9419703 TI - Regression analysis of correlated receiver operating characteristic data. PMID- 9419704 TI - Issues in combining independent estimates of the sensitivity and specificity of a diagnostic test. PMID- 9419705 TI - Regression methods for meta-analysis of diagnostic test data. PMID- 9419706 TI - Image-enhancing agents. PMID- 9419707 TI - Contrast media and radiopharmaceutical agents: regulatory issues. PMID- 9419708 TI - Single-photon tracers for single-photon emission computed tomography. PMID- 9419709 TI - Positron emission tomography: radiochemistry. PMID- 9419710 TI - In vivo biochemistry, pharmacology, and physiology. PMID- 9419711 TI - Technology assessment. PMID- 9419712 TI - Display, picture archiving and communicating systems, and teleradiology. PMID- 9419713 TI - Informatics. PMID- 9419714 TI - Medical image perception. PMID- 9419715 TI - X-ray imaging: projection radiography. PMID- 9419716 TI - X-ray generation and image capture: monoenergetic X-ray sources. PMID- 9419717 TI - Ultrasound: physics and instrumentation. PMID- 9419718 TI - Ultrasound: clinical applications. PMID- 9419719 TI - Single-photon imaging. PMID- 9419720 TI - Positron emission tomography. PMID- 9419721 TI - Interventional magnetic resonance imaging, computed tomography, and ultrasound. PMID- 9419722 TI - Vascular intervention. PMID- 9419723 TI - Magnetic resonance imaging: instrumentation. PMID- 9419724 TI - Vascular magnetic resonance imaging. PMID- 9419725 TI - Functional magnetic resonance imaging. PMID- 9419726 TI - Nuclear magnetic resonance spectroscopy. PMID- 9419727 TI - Computed tomography: physics and instrumentation. PMID- 9419728 TI - Computed tomography: applications. PMID- 9419729 TI - Bioelectric and biomagnetic imaging. PMID- 9419730 TI - New imaging technologies. PMID- 9419731 TI - Imaging of the central nervous system. PMID- 9419732 TI - Imaging of the heart. PMID- 9419733 TI - Imaging of the breast. PMID- 9419734 TI - Imaging of the chest. PMID- 9419735 TI - Imaging of the gastrointestinal tract. PMID- 9419736 TI - Imaging of the genitourinary system. PMID- 9419737 TI - Pediatric radiology. PMID- 9419738 TI - Imaging of the musculoskeletal system. PMID- 9419739 TI - The yolk sac theory: closing the circle on why diabetes-associated malformations occur. AB - OBJECTIVE: The purpose of this article is to examine the role of yolk sac failure during organogenesis in the development of diabetes-associated embryopathy. METHODS: The current literature regarding congenital malformations in diabetic pregnancies was reviewed to elucidate the precise role of the yolk sac in embryonic development and the relation between yolk sac injury and embryopathy. RESULTS: We and others have demonstrated that hyperglycemia produces a teratogenic effect during organogenesis. In addition, we have shown that the yolk sac appears to be the target site of injury induced by hyperglycemia. We have also presented evidence that cell membrane dysfunction leads to failed vitelline vessel formation and that arachidonic acid supplementation prevents many of the morphologic and biochemical alterations observed under hyperglycemic conditions. CONCLUSIONS: These data strongly support the teratogenic effect of hyperglycemia, the arachidonic acid deficiency state, the resultant maldevelopment of vitelline vessels, and the ability to prevent these changes by arachidonic acid supplementation. These studies have made significant inroads in explaining why diabetes-associated anomalies occur, and suggest a potential future role for prophylaxis against these organogenetic malformations using dietary polyunsaturated fatty acid supplementation. PMID- 9419740 TI - Gene expression of atrial natriuretic factor in ovine fetal heart during development. AB - OBJECTIVE: The present study quantified the abundance of atrial natriuretic factor (ANF) messenger RNA (mRNA) and determined the developmental pattern of ANF gene expression in the four cardiac chambers of the ovine fetus during the last two-thirds of gestation. METHODS: Twenty-one fetuses from 13 time-dated pregnant ewes at gestational ages of 60-145 days were used for this study. Total RNA from fetal atria and ventricles was extracted and ANF mRNA was analyzed by Northern blotting. The ANF mRNA signal was quantified by light densitometry. The abundance of ANF mRNA in the cardiac chambers across gestational ages was analyzed by linear regression analysis and one-way analysis of variance. RESULTS: Atrial natriuretic factor mRNA was much more abundant in the atria than in the ventricles of all fetuses at each gestational age studied. Atrial ANF mRNA levels were lowest in the younger fetuses at 60 days and increased with advancing gestation. Ventricular ANF mRNA levels were highest in fetuses at 60 days and decreased to almost nondetectable levels near term. No difference in ANF mRNA abundance was noted between the right and left atria or the right and left ventricles at each gestational age. CONCLUSION: A developmental pattern of ANF gene expression is demonstrated in the ovine fetal heart during the last two thirds of gestation. This pattern shows that atrial ANF mRNA abundance increases while ventricular abundance decreases as the fetus matures. Expression of the ANF gene in the fetal period may be regulated developmentally or induced by cardiovascular changes in utero. PMID- 9419741 TI - Vasodilator effects of parathyroid hormone, parathyroid hormone-related protein, and calcitonin gene-related peptide in the human fetal-placental circulation. AB - OBJECTIVE: The purpose of our study was to determine the vasoactivity of calcitonin gene-related peptide (CGRP), parathyroid hormone (PTH), and parathyroid hormone-related protein (PTHrP) in the human fetal-placental circulation in vitro. METHODS: Dually perfused placental cotyledons from term pregnancies were used in this study. RESULTS: Calcitonin gene-related peptide, PTHrP (both 10(-10)-10(-6) mol/L), and PTH (10(-8)-10(-6) mol/L) demonstrated a significant concentration-dependent vasodilator effect (P = .0007, P = .0172, P = .0063, respectively), following preconstriction with a thromboxane mimetic U46619. The CGRP-1 receptor inhibitor CGRP8-37 (10(-6) mol/L) significantly inhibited (P = .0131) the CGRP-induced vasodilator effect, while the nitric oxide synthesis inhibitor n-nitro-l-arginine showed no inhibitory effect. CONCLUSIONS: These results demonstrate the vasodilator effects of CGRP, PTH, and PTHrP in the human fetal-placental circulation. Calcitonin gene-related peptide and PTHrP were of equal potency, and both were approximately 100 times more potent than PTH. This study also suggests the CGRP may exert its vasodilator effect through two classes of receptors in the human placenta and may do so independently of nitric oxide. PMID- 9419742 TI - Magnesium sulfate treatment decreases N-methyl-D-aspartate receptor binding in the rat brain: an autoradiographic study. AB - OBJECTIVE: We determined the effect of peripherally administered magnesium sulfate on N-methyl-D-aspartate (NMDA) receptor binding capacity in various regions of the rat brain. METHODS: Three separate experiments were performed. 1) Six rats were injected intraperitoneally with 270 mg/kg of magnesium sulfate, followed by 27 mg/kg every 20 minutes for 4 hours; controls (n = 6) received saline. 2) Six rats received intraperitoneal injections of magnesium sulfate (270 mg/kg) every 4 hours for 24 hours, while six received saline. 3) Six rats received intraperitoneal magnesium sulfate (270 mg/kg) every 12 hours for a total of 2 weeks, and six received saline. Rats were subsequently perfused and sacrificed, and their brains were dissected, rinsed, and frozen. Cryostat sections were taken, labeled by in vitro [3H]-CGP 39653, assayed autoradiographically, and mounted on Ultrofilm for 4 weeks. Optical density measurements of binding on each section were performed using an image analyzing system. Eleven brain regions were sampled: 1, 2) frontal and occipital cortex; 3 7) hippocampus--CA-1, CA-3, stratum radiatum, stratum oriens, dentate gyrus; 8) thalamus; 9) hypothalamus; 10) caudate nucleus; and 11) cerebellum. RESULTS: The NMDA receptor binding density in the hippocampus was significantly higher than in all other brain regions in all three experiments. In experiment 1, there was no significant effect on NMDA receptor binding. However, prolonged systemic administration of magnesium sulfate for 24 hours resulted in significantly reduced [3H]-CGP binding in all brain regions sampled. After chronic magnesium sulfate administration (2 weeks), the [3H]-CGP binding was still reduced in the cortex and some regions of the hippocampus; however, there was no significant change in other regions. CONCLUSIONS: Peripheral treatment with magnesium sulfate results in a significant reduction in the NMDA receptor binding capacity in the rat brain. These results support the hypothesis that magnesium central activity is mediated, at least in part, via the NMDA receptor. PMID- 9419743 TI - Mechanism of cytokine stimulation of prostaglandin biosynthesis in human decidua. AB - OBJECTIVE: The purpose of this study was to determine the mechanism of action of the cytokines tumor necrosis factor-alpha (TNF alpha) and interleukin-1 beta (IL 1 beta) on the stimulation of prostaglandin (PG) production in human decidua. METHODS: Decidual cells from term placentas were grown in culture until confluent. Incubations were performed with TNF alpha or IL-1 beta, and with cycloheximide, actinomycin D, arachidonic acid, or acetylsalicylic acid (ASA). Prostaglandin E2 was measured by radioimmunoassay and cellular protein determined. RESULTS: The concentration-related stimulation of decidual PGE2 production by IL-1 beta and TNF alpha was completely abrogated by cycloheximide and actinomycin D treatment. Although arachidonic acid alone stimulated decidual PGE2 biosynthesis, the addition of IL-1 beta or TNF alpha consistently augmented this effect. Both cytokines induced recovery of PGE2 biosynthesis from ASA pretreatment more rapidly than controls. CONCLUSIONS: Interleukin-1 beta and TNF alpha both act on decidual PG biosynthesis in a manner requiring new protein synthesis. In combination with our other results, this suggests that IL-1 beta and TNF alpha act to induce PG endoperoxide synthase activity. PMID- 9419744 TI - Assessments of fetal swallowed volume: tracer disappearance versus esophageal flow. AB - OBJECTIVE: We sought to compare fetal swallowed volume determinations simultaneously by two techniques--amniotic fluid (AF) tracer disappearance and esophageal flow probe measurements. METHODS: Six ovine fetuses (129 +/- 1 days) were chronically prepared with a thoracic esophageal flow probe and vascular and two AF catheters. 125I-labeled albumin was injected into the AF cavity, and samples were withdrawn at timed intervals for 8 hours. The AF volume was calculated by time-0 extrapolation of the semilog of 125I disappearance. Tracer determined swallowed volume was calculated as the product of AF volume and the slope of isotope disappearance. Flow probe measurement of swallowed volume was determined by computer integration of calibrated flow probe velocity recordings. RESULTS: The AF volume averaged 805 +/- 168 mL. The isotope disappearance rate from the AF was 2.8 +/- 0.4%/hour. Average tracer-determined swallowed volume (547 +/- 113 mL/day) was greater than flow probe volume (366 +/- 81 mL/day), although these values were not significantly different. However, when corrected for estimated swallowed lung fluid, tracer-determined volume was significantly greater than flow probe volume (P < .05). CONCLUSION: Ovine fetal swallowed volume determinations by AF tracer techniques are greater than those determined by esophageal flow probe measurements. PMID- 9419745 TI - Studies on the placental transfer of cell-free human immunodeficiency virus and p24 antigen in an ex vivo human placental model. AB - OBJECTIVE: We studied whether the human placenta has the structural integrity to impede transplacental passage of cell-free human immunodeficiency virus (HIV)-1 or p24 antigen from the maternal to the fetal circulation. METHODS: Nine term human placentas from uncomplicated vaginal or cesarean section deliveries were studied ex vivo with a placental perfusion apparatus to determine whether cell free HIV-1 at 200-2000 tissue culture infectious dose (TCID50/mL) would pass to the fetal circulation. Passage of virus or p24 was assessed by infectivity titration and/or p24 antigen capture enzyme immunoassay. RESULTS: Infectious HIV 1 was not detected in any of the fetal perfusate samples taken periodically during experiments. Low concentrations of HIV-1 p24 antigen, however, were detected in fetal perfusate samples from three placentas. CONCLUSIONS: The term human placenta effectively impedes passage of cell-free HIV-1 from the maternal to the fetal circulation. However, it may be permeable to passage of p24 antigen. PMID- 9419746 TI - The relationship between oxytocin, phosphoinositide-specific phospholipase C, and phasic myometrial contractions. AB - OBJECTIVES: Oxytocin-stimulated uterine contractions are associated with repetitive cycles of elevated cytosolic calcium, i.e., cytosolic calcium oscillations. The studies in this report were performed to test the hypothesis that phosphoinositide-specific phospholipase C (PI-PLC) is an important component of the myometrial intracellular oscillator. METHODS: In vitro isometric contraction studies were performed using longitudinal strips of myometrium from nonpregnant, adult Sprague-Dawley rats. Cumulative dose-response studies were performed using oxytocin and aluminum fluoride with and without 2-nitro-4 carboxyphenyl-N, N-diphenylcarbamate (NCDC), an inhibitor of PI-PLC. RESULTS: Stimulation of G-proteins coupled to PI-PLC with aluminum fluoride resulted in a significant increase in phasic myometrial contractions comparable to those produced by oxytocin. Inhibition of PI-PLC with NCDC resulted in significant suppression of oxytocin- and aluminum fluoride-stimulated myometrial contractions. In contrast, doses that suppressed agonist-stimulated contractions had only a minimal effect on KCl-stimulated tonic myometrial contractions. CONCLUSIONS: These studies provide significant support for the novel hypothesis that PI-PLC is an important component of the agonist-stimulated cytosolic calcium oscillator that generates phasic myometrial contractions. PMID- 9419747 TI - Positive serum screening for fetal Down syndrome does not predict adverse pregnancy outcome in absence of fetal aneuploidy. AB - OBJECTIVE: The purpose of this study was to determine whether false-positive maternal serum screening for fetal Down syndrome is predictive of poor pregnancy outcome. METHODS: The pregnancy outcomes of 99 women having positive serum screening for fetal Down syndrome (study group)--based upon maternal serum alpha fetoprotein (MSAFP), unconjugated estriol (uE3), hCG, and maternal age--were compared to the outcomes of matched control patients having negative serum screening results (control group). The outcome indices analyzed were fetal death, intrauterine growth retardation (IUGR), preeclampsia, and fetal anomalies. RESULTS: Between the study group and the control group, there were no statistically significant differences in pregnancy outcome with respect to fetal death, IUGR, preeclampsia, or fetal anomalies. CONCLUSIONS: Our findings demonstrate no apparent increase in the adverse perinatal outcomes analyzed in women having unexplained positive serum screening for fetal Down syndrome. Although further investigation is needed, these results provide no evidence to support increased antepartum surveillance in such patients. PMID- 9419749 TI - Whole endometrial fragments form characteristics of in vivo endometriosis in a mesothelial cell co-culture system: an in vitro model for the study of the histogenesis of endometriosis. AB - OBJECTIVE: We have previously reported on the effects of tumor necrosis factor alpha (TNF-alpha) and interleukin-1 (IL-1) on the adhesion of human endometrial stromal cells to peritoneal mesothelial cells in vitro. The relevance of this co culture system to in vivo endometriosis remains to be established. We contrasted endometrial fragments with endometrial stromal cells to determine their relevance. METHODS: Human mesothelial cells were isolated from peritoneal fluid from four normal women via Ficoll-Paque gradient centrifugation at 400 x g for 30 minutes and grown in M199 medium containing epidermal growth factor (10 ng/mL) and 10% fetal calf serum. The homogeneous cells were cultured on collagen-coated plates until a monolayer formed. Endometrial stromal cells or whole endometrial fragments, isolated from endometrial tissue of four normal women, were put on the mesothelial monolayer and cultured at 37 degrees C for 24 days. After fixation, the samples were incubated with monoclonal antibody to epithelial membrane antigen, cytokeratin, and vimentin, and processed with standard immunohistochemical techniques. RESULTS: Observation under phase contrast microscopy revealed that, in this co-culture system, whole endometrial fragments demonstrated characteristic morphology of in vivo endometriosis, whereas isolated endometrial stromal cells did not. CONCLUSION: Whole endometrial fragments, but not isolated endometrial stromal cells, form morphologically characteristic structures similar to in vivo endometriosis in this co-culture system. It is hoped that this system might be useful for studying certain aspects of the histogenesis of endometriosis. PMID- 9419748 TI - Tumor necrosis factor-alpha inhibits ovulation and steroidogenesis, but not prostaglandin production in the perfused rat ovary. AB - OBJECTIVE: We tested the null hypothesis that tumor necrosis factor-alpha (TNF alpha) does not decrease ovulation, estradiol and progesterone production, or prostaglandin (PG) E2, PGF2alpha, or 6 keto-PGF1alpha production in the open bursa rat ovarian perfusion model. METHODS: Experimental animals were controlled for age, weight, litter, and pregnant mare's serum gonadotropin (PMSG) aliquot. Female Sprague-Dawley rats, 26-27 days old, were injected with 25 IU PMSG. Forty eight hours later, the right ovary was dissected, the bursa removed, and the specimen placed in the perfusion chamber with defined media. Luteinizing hormone and isobutylmethylxanthine were given as an ovulatory trigger. Test perfusions also received TNF-alpha in 0.8-nmol/L, -pmol/L, and -fmol/L doses. Samples were collected at 0, 1, 3, 5, 7, 10, and 20 hours. Ovulations were counted at 20 hours. Steroids and PGs were measured. RESULTS: The addition of TNF-alpha to the rat ovarian perfusion model resulted in a dose-dependent decrease in ovulations (mean +/- standard deviation): 16.14 +/- 6.2 in controls (n = 7) versus 2.38 +/- 3.4 with TNF-alpha 0.8 nmol/L (n = 7), and 4.3 +/- 1.5 with TNF-alpha 0.8 pmol/L (n = 3), both P < .001. Tumor necrosis factor-alpha also inhibited estradiol (P < .005) and progesterone production (P < .05) throughout, but produced no significant changes in PG production. CONCLUSIONS: Tumor necrosis factor-alpha inhibits ovulation in a dose-dependent fashion, and inhibits estradiol and progesterone production without altering PG production in the open bursa rat ovarian perfusion model. PMID- 9419750 TI - Penetration of human spermatozoa through the zona pellucida of nonviable human oocytes. AB - OBJECTIVE: The purpose of this study was to evaluate the penetrating capacity of human sperm through the intact zonae of nonviable human oocytes and the influence of oocyte storage conditions and gamete coincubation times on penetration rates. METHODS: Immature oocytes were obtained from surgically removed ovarian tissue and from in vitro fertilization (IVF) and subjected to four storage conditions: 1) storage at 4 degrees C for up to 48 hours in culture medium (refrigeration, n = 53), 2) cryopreservation in 1.5 mol/L propanediol with storage at -196 degrees C (PrOH; n = 49), 3) cryopreservation in 2.0 mol/L dimethylsulfoxide with storage at -70 degrees C (DMSO; n = 20), and 4) storage in a hypertonic salt solution at 4 degrees C (salt; n = 30). Semen was obtained from fertile donors (n = 7) and after a wash and swim-up, samples were adjusted to 500,000 motile sperm/mL. Zonae and sperm were coincubated in 100-microL drops under oil, and penetration was assessed at 5 and 20 hours. Penetration was evidenced by visualization of at least one sperm head in the perivitelline space at 200x magnification. RESULTS: Zona penetration rates were 50.9, 32.7, 23.3, and 10.0% for refrigeration, PrOH, salt, and DMSO storage, respectively (P < .004). There was a significantly higher number of sperm in the perivitelline space after 20 hours' coincubation (3.8 +/- 0.2) compared to 5 hours (0.8 +/- 0.1) (P < .0006). This represented an increase in the penetration rate from 31.2% (34 of 109) at 5 hours to 37.6% (41 of 109) at 20 hours, although the difference was not significant. CONCLUSIONS: These results demonstrate that the zona pellucida of nonviable human oocytes can be used in a zona penetration assay and that refrigerated and PrOH-frozen intact zonae have the highest rates of penetration. The decrease in penetrability seen in DMSO and salt-stored oocytes may be due to alterations occurring in the zona pellucida. PMID- 9419751 TI - Enhanced post-receptor insulin effects in women following dehydroepiandrosterone infusion. AB - OBJECTIVE: We hypothesized that intravenous dehydroepiandrosterone (DHEA) would decrease insulin resistance in normal and insulin-resistant women. METHODS: Five insulin-resistant women diagnosed as having polycystic ovaries (PCO) with elevated testosterone and normal dehydroepiandrosterone sulfate (DHEAS) with amenorrhea were recruited. Obese controls (OC) with normal menses and normal testosterone and DHEAS were recruited and matched to each PCO woman for age and weight. The PCO women had a mean testosterone of 3.2 +/- 0.4 nmol/L, fasting serum insulin level of 330 +/- 55 pmol/L, and DHEAS level of 3.4 +/- 1.3 mumol/L. An oral glucose tolerance test (OGTT) was performed at 8 AM after an overnight fast. A DHEA infusion (1 mg/hour for 17 hours) was begun at 6 PM and continued until the completion of the second OGTT performed the following morning at 8 AM. T-lymphocytes were drawn at 8 AM each morning. RESULTS: The DHEA infusion had no significant effect on any of the in vivo indices of insulin sensitivity, ie, basal and OGTT insulin, C-peptide, and ratios of insulin/glucose. In vitro, DHEA significantly increased insulin binding to T-lymphocytes of PCO women but caused no significant change in OC women. There was, however, marked enhancement of T lymphocyte pyruvate dehydrogenase (PDH) activities in both groups of study subjects following DHEA. CONCLUSION: We conclude that a 17-hour infusion of DHEA enhanced T-lymphocyte insulin binding and PDH activity while producing no detectable improvements in in vivo indices of insulin sensitivity. PMID- 9419752 TI - The effect of progestins on behavioral stress responses in postmenopausal women. AB - OBJECTIVE: We assessed the effects of progestin when added to estrogen on the adaptive patterns to provoked stress in postmenopausal women. METHODS: Fourteen postmenopausal women were randomized to receive either a transdermal estrogen patch (TE2) (n = 7) for 6 weeks or TE2 with added medroxyprogesterone acetate (10 mg) (TE2/MPA) (n = 7) for the last 10 days of the 6-week regimen. Behavioral stress tests were administered to each group, with measurements of biophysical and neuroendocrine responses. In a crossover fashion, after each group received the first treatment and testing, treatment was continued for another 6 weeks with the alternate regimen, at which time another stress test was administered. Responses to stress in the two treatment groups were compared to each other and to established placebo responses. RESULTS: Biophysical responses in the TE2 group were significantly blunted compared to both TE2/MPA and placebo responses (P < .05). Without MPA treatment, there were significantly blunted speech (P < .05) and cold pressor (P < .01) blood pressure responses. With added progestin, there was a greater systolic blood pressure response (P < .01) compared with estrogen alone. Both groups (TE2 and TE2/MPA) had blunted and nonsignificant responses of ACTH and cortisol upon testing, whereas the placebo group showed a significant response (P < .01). Plasma norepinephrine responses, however, were significantly blunted after TE2, compared with the increased responses observed with both TE2/MPA and placebo (P < .01). CONCLUSION: Although estrogen significantly reduces behaviorally induced stress reactivity in postmenopausal women, certain doses of progestin administration may blunt this effect. PMID- 9419753 TI - Hypothalamic amenorrhea and hidden nutritional insults. AB - OBJECTIVE: We examined whether abnormal nutrition is an associated event in idiopathic hypothalamic amenorrhea. METHODS: Eighteen amenorrheic subjects were compared to 36 normal controls using endocrine, nutritional, and psychological evaluations. RESULTS: Controls were closer to their ideal weight (97.2 versus 89.7%; P < .05) than amenorrheics despite similar ages and heights. Amenorrheics denied eating disorders; however, our evaluation showed more eating disorders (55 versus 26%; P < .05), higher scores on a scale of eating behavior (22.28 versus 10.36; P < .001), twice as much fiber intake (26.14 versus 14.69 g/day) and less fat intake (20.7 versus 27.10 g/day) (P < .001), more aerobic activity (85 versus 58%) despite expending fewer calories per day (2303.7 versus 2576.7 kcal/day; P < .05), and similar caloric intake. CONCLUSIONS: Significant abnormalities suggest that a greater percentage of hypothalamic amenorrhea occurs on a nutritional basis than previously suspected and should be searched for in depth. PMID- 9419754 TI - Epidermal growth factor enhances secretion of the ovarian tumor-associated cancer antigen CA125 from the human amnion WISH cell line. AB - OBJECTIVE: We studied the relation between epidermal growth factor (EGF)/epidermal growth factor receptor (EGFR) and CA125 production in WISH cells. METHODS: We investigated quantitatively and immunohistochemically EGF-stimulated CA125 release from WISH cells and the effect of EGF on CA125 phosphorylation. RESULTS: Immunohistochemical staining demonstrated that CA125 and EGFR expression on the plasma membrane of the WISH cells was closely correlated with cell density. The WISH cell monolayers (day 4) stained for CA125 in both the cytoplasm and plasma membrane. By day 8, cells began to form clumps in the surrounding monolayer that were positive for membrane-associated CA125 and EGFR, while the monolayer was almost negative for both molecules. Four-day and 8-day cells exposed to EGF demonstrated a loss of both CA125 and EGFR staining. Epidermal growth factor increased the secreted CA125 levels by 50% in day-4 cells but had no effect on day-8 cells. CA125 from WISH cells was phosphorylated, and EGF further enhanced this phosphorylation. PMID- 9419756 TI - The impact of primary care on academic medicine. PMID- 9419755 TI - Estrogen mustard induces cell cycle arrest of human epithelial ovarian cancer cell lines. AB - OBJECTIVE: Pharmacologic disruption of microtubule function may provide effective therapy for advanced epithelial ovarian cancer, as has been observed in clinical trials using taxol. However, the limited availability of taxol and taxol's side effects emphasize a need to develop alternative antimicrotubule agents. Estramustine (EM) inhibits binding of microtubule-associated proteins (MAPs) to microtubules, promotes microtubule disassembly, disrupts spindle formation, and induces metaphase arrest in human prostate carcinoma and glioma cells in culture. We studied the effect of EM on DNA synthesis and on the cell cycle in four human ovarian carcinoma cell lines and examined the cell lines for evidence of MAP-like immunoreactivity. METHODS: The effect of EM on DNA synthesis and on the cell cycle was determined using [3H]thymidine incorporation assays and flow cytometry, respectively. Microtubule-associated protein-like immunoreactivity was determined using monoclonal antibodies directed against MAP 1A, MAP 1B, and MAP 2(2A + 2B) for Western analysis after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. RESULTS: We demonstrated a dose-dependent inhibitory response to EM in BIXLER, DK2NMA, and SKOV3. BIX3 showed a dose-dependent inhibitory response to EM concentrations from 25 micrograms/mL to 100 micrograms/mL, but a stimulatory response at 10 micrograms/mL. Estramustine inhibited exponentially growing cells by causing mitotic arrest with subsequent accumulation of cells in G2/M phase of the cell cycle in all four cell lines. We found MAP 1A, MAP 1B, and MAP 2-like immunoreactivity in all four cells lines studied. CONCLUSIONS: These results are consistent with a MAP-microtubule mechanism of action for EM in ovarian carcinoma cells and provide reason to conduct further study of EM for potential use in the treatment of human epithelial ovarian cancer. PMID- 9419757 TI - The gynecologist and cardiovascular disease: a window of opportunity for prevention. AB - OBJECTIVE: We present current concepts and assess the quality of information available for the prevention of cardiovascular disease in women. METHODS: This article reviews research bearing on the prevention of cardiovascular disease in women, with particular attention to modifiable risk factors. We describe the magnitude of the problem and assess the quality of the data with respect to the classic risk factors. The concept is emphasized that changes at menopause, states of endocrine aberration, and benefits and risks of hormone substitution and oral contraception must be understood in conjunction with all other potentially modifiable and nonmodifiable risk factors. CONCLUSIONS: Primary care physicians, especially obstetrician/gynecologists, have a pivotal role to play in the reduction of this disease. Behavior modification is the key to integrating prevention into the regular annual visit. PMID- 9419758 TI - The basis and evidence of a role for the ovarian renin-angiotensin system in health and disease. AB - OBJECTIVE: We reviewed the evidence for an intrinsic ovarian renin-angiotensin system (OVRAS), highlighting potential diverse signaling in this system through different bioactive angiotensin peptides, their specific receptors, and second messengers. In addition, sites of action for OVRAS in the regulation of ovarian function in health and disease were reviewed. DATA SOURCES: We used published journals and abstracts from national scientific meetings. Current developments in the renin-angiotensin field are historically set. STUDY SELECTION: One hundred referenced articles provided studies on renin-angiotensin systems in mammalian species, including humans. DATA ABSTRACTION: Interpretation of the reviewed publication was in line with the original authors' conclusions and statistical analysis. DATA SYNTHESIS: Techniques in molecular biology, biochemistry, and immunohistochemistry have identified an OVRAS in mammalian species. Ovarian tissues contain all the elements for the production of angiotensin, including prorenin/renin, angiotensinogen, and angiotensin-converting enzyme. In addition, angiotensin II is present in ovarian compartments, and receptors for angiotensin II are demonstrated on specific ovarian cells. Angiotensin II is implicated to play a role in ovulation, steroidogenesis, follicular atresia, and hyperandrogenic syndromes. CONCLUSIONS: The newly identified OVRAS may have important actions in the ovary that range from regulation of ovulation to ovarian dysfunction, such as hyperandrogenic syndromes in women. In this respect, the OVRAS is a putative paracrine/autocrine regulator in the ovary, and pharmacologic regulation of the OVRAS may provide new methods for the management of fertility and reproduction. PMID- 9419759 TI - Molecular analysis to assign parental origin and distinguish de novo i(21q) from t(21q21q) in two Down syndrome fetuses. AB - OBJECTIVE: We sought to determine the origin of two prenatal cases of chromosome 21 rearrangements not amenable to clarification by conventional cytogenetic methodology. METHODS: Hypervariable repeat polymorphisms (chromosome 21) were used to determine the type of structural rearrangement and the parental origin of the rearranged chromosome. The repeats used were highly polymorphic and located very close to the centromere; thus, the likelihood of differences among the parental alleles and overall informativeness were increased. RESULTS: The rea(21q21q) chromosomes were identified as a Robertsonian translocation in one fetus and an isochromosome in the other. The extra chromosome material was found to be maternal in origin in both cases. CONCLUSION: The ability to clarify the origin of abnormal chromosomal rearrangements provides valuable information concerning possible mechanisms of aneuploidy, as well as clinical data that may have an impact in assessing a patient's risk for abnormal offspring. PMID- 9419760 TI - Modulators of prostaglandin E2 synthesis in human amnion. AB - OBJECTIVE: The purpose of these studies was to determine the effects of the essential fatty acid, linoleic acid, and the commonly used non-steroidal anti inflammatory agents, aspirin and acetaminophen, on the rate of prostaglandin (PG) biosynthesis by human amnion cells. METHODS: Amnion cells were isolated from term, normal pregnancies and grown to confluence. Cells were incubated with control or medium containing 100 mumol/L linoleic acid. Cells were also incubated with control medium or medium containing 10 or 100 micrograms/mL aspirin or acetaminophen. RESULTS: Following an initial delay, amnion cells exposed to linoleic acid exhibited a significant increase in PGE synthesis. Both aspirin and acetaminophen in clinically relevant concentrations had a significant inhibitory effect on amnion cell PGE synthesis. CONCLUSIONS: Linoleic acid has a stimulatory effect and aspirin and acetaminophen have an inhibitory effect on PGE synthesis in human amnion cells in culture. We speculate that dietary habits, supplement ingestion, and over-the-counter drug use may affect amnion cell PG production. In view of the potential importance of intrauterine PG production in normal and abnormal labor, further study in this area is indicated. PMID- 9419761 TI - Platelet activating factor-acetylhydrolase activity following chorionic villus sampling and amniocentesis. AB - OBJECTIVE: Platelet activating factor (PAF) is essential for embryonic development and is a potent vasodilator. It increases vascular permeability and stimulates prostaglandin E2 (PGE2) production. Platelet activating factor acetylhydrolase (PAF-AH), the enzyme that degrades PAF, is synthesized by decidual macrophages. The aim of this study was to test the hypothesis that chorionic villus sampling (CVS) and/or amniocentesis might cause an increase in maternal PAF-AH activity. METHODS: Maternal plasma PAF-AH activity was evaluated before and after genetic amniocentesis (N = 13) and transcervical CVS (N = 29). A control group (N = 9) was evaluated to study the effects of venipuncture. RESULTS: Chorionic villus sampling caused a significant elevation in PAF-AH activity (P < .0005). No changes were noted in PAF-AH activity in the amniocentesis or the control group. CONCLUSIONS: Chorionic villus sampling causes subclinical release of PAF-AH, possibly from the decidual macrophages. Increased PAF-AH activity might result in decreased PAF levels, which might lead to vasoconstriction in the placental circulation due to lack of the vasodilator effects of PAF and possibly PGE2. This mechanism might explain the increased risk for fetal limb reduction noted with CVS performed at very early gestational ages. PMID- 9419762 TI - Pregnancy outcome following ultrasound-detected fetal cardiac activity in women with a history of multiple spontaneous abortions. AB - OBJECTIVE: To determine whether women with two or more previous spontaneous abortions of unknown etiology, by conventional testing criteria, have a different rate of subsequent fetal loss than controls after ultrasonic documentation of fetal cardiac activity. METHODS: Medical records were reviewed from 185 women with spontaneous abortion of unknown etiology. Of these women, 91.9% were found to have evidence of cellular immunity to trophoblast and were treated with progesterone for immunosuppression. Ultrasound evaluation was obtained at 5-6 weeks' gestation to document fetal cardiac activity. A control group of 63 women was also studied. All women were followed for pregnancy outcome. RESULTS: A total of 248 pregnancies were identified from the 185 study patients with multiple spontaneous abortions. Fetal cardiac activity was visualized by ultrasound in 209 pregnancies from 171 study subjects; of these, the outcomes of 208 pregnancies were known. The rate of spontaneous abortion after ultrasound documentation of fetal cardiac activity was 22.7%. Neither maternal age nor number of previous losses was associated with an increased incidence of spontaneous abortion following documentation of fetal cardiac activity. The rate of spontaneous abortion in the control group after documentation of fetal cardiac activity was 3.3%. CONCLUSION: These data may help clinicians give couples who have experienced recurrent pregnancy loss a more realistic prognosis for pregnancy success once fetal cardiac activity has been confirmed. PMID- 9419763 TI - Expression of engineered human 17 beta-estradiol dehydrogenase in a prokaryotic system. AB - OBJECTIVE: 17 beta estradiol dehydrogenase (17 beta DH) is a model for pyridine dependent steroid-converting enzymes. To define the structural and functional parameters of 17 beta DH, we created an expression system for production of abundant, homogeneous enzyme. METHODS: A full-length 17 beta DH cDNA clone was engineered into the inducible expression vector pMON 5839. After induction of plasmid-bearing Escherichia coli JM109 cells, the authenticity of the recombinant human placental 17 beta DH (r17 beta DH) was evaluated. RESULTS: Protein electrophoresis and Western blot analysis confirmed the immunologic identity of r17 beta DH with native human placental enzyme. The amino acid sequence, enzyme activity, Vmax, K(m), and kcat of r17 beta DH matched that of the native enzyme. CONCLUSION: Prokaryotic cell lines offer the opportunity to create an unlimited supply of recombinant human placental 17 beta DH without the expense and time commitment of baculoviral or eukaryotic cell lines. We are now able to use r17 beta DH and its mutants to elucidate the mechanisms of action of this class of enzyme. PMID- 9419764 TI - Comparison of estimates of insulin sensitivity in pre- and postmenopausal women using the insulin tolerance test and the frequently sampled intravenous glucose tolerance test. AB - OBJECTIVE: We assessed insulin sensitivity in women comparing the insulin tolerance test (ITT) with the intravenous glucose tolerance test with frequent sampling and computer modeling (FSIVGTT) and evaluated the effects of hormonal therapy in postmenopausal women using both methods. METHODS: This prospective study tested 18 premenopausal women and ten postmenopausal women randomized to receive either estrogen alone or estrogen with a sequential progestin for 6 months at a menopause research clinic. All subjects received an ITT and an FSIVGTT within 48-72 hours of each other in random sequence. Postmenopausal women were then randomized to receive either 0.625 mg conjugated equine estrogen for 6 months or 0.625 mg conjugated equine estrogen with medroxyprogesterone acetate, 10 mg, for 10 days each month for 6 months. Both the ITT and the FSIVGTT were repeated following hormonal therapy at 2 and 6 months. Plasma insulin and glucose were measured; insulin sensitivity was calculated after the ITT (Kitt) and the FSIVGTT (Si) at each visit in each group. RESULTS: A close correlation was found between Kitt and Si values at initial testing in both pre- and postmenopausal women and following both types of hormonal therapy (r = 0.76 for all tests, P < .001). A reduction in insulin sensitivity was observed in postmenopausal compared to premenopausal women; this occurred in five of ten postmenopausal women using the Kitt measurement and in four of ten women using Si. Estrogen replacement had a beneficial effect on insulin sensitivity. While Kitt increased by 24.2 +/- 9.6% (P < .05), the increase in Si (6.7 +/- 18%) was not significant because of the variability with this measurement. An attenuation in insulin sensitivity was seen with added progestin. Kitt values decreased by 17.7 +/- 7.7% and Si values by 31.9 +/- 12%. Similar findings were noted at 2 and 6 months. CONCLUSIONS: The ITT and FSIVGTT provide quantitatively similar information regarding insulin sensitivity in healthy women. A mild degree of insulin resistance appears to be present in some healthy postmenopausal women. Estrogen appears to improve insulin sensitivity, while added progestin may attenuate this beneficial effect. PMID- 9419765 TI - Affinity labeling in the presence of the reduced diphosphopyridine nucleotide NADH identifies peptides associated with the activities of human placental 3 beta hydroxy-delta 5-steroid dehydrogenase/isomerase. AB - OBJECTIVE: We sought to identify peptides associated with activity in the primary structure of human placental 3 beta-hydroxy-delta 5-steroid dehydrogenase/isomerase (3 beta-HSD/isomerase). METHODS: Purified human placental 3 beta-HSD/isomerase was affinity-radioalkylated by 2 alpha-bromo [2' 14C]acetoxyprogesterone (2 alpha-[14C]BAP) in the presence or absence of the reduced diphosphopyridine nucleotide, NADH. NADH protected both 3 beta-HSD and isomerase from inactivation by 2 alpha-[14C]BAP. Tryptic peptides of unprotected and NADH-protected radioalkylated enzyme were purified by high-pressure liquid chromatography. The amino acid sequence of each radiolabeled peptide was determined and localized within the cDNA-derived primary structure of the enzyme. RESULTS: According to the sequence analyses, NADH shifted radioalkylation by 2 alpha-[14C]BAP away from the Arg-250 peptide (251GQFYYISDDTPHQSYDNLNYTLSK274) and toward the Lys-135 tryptic peptide (136EIIQNGHEEEPLENTWPAPYPHSK159). Based on amino acid analysis to quantitate radioactivity incorporated per nmol peptide, NADH decreased the radiolabeling of His262 in the Arg-250 peptide by 8.2-fold. His142 in the Lys-135 peptide was radiolabeled by 2 alpha-[14C]BAP only in the presence of NADH. CONCLUSIONS: We have previously reported that the substrate pregnenolone blocks the inactivation of 3 beta-HSD by 2 alpha-[14C]BAP through the protection of His262 in the Arg-250 peptide. Protection by NADH against the inactivation of isomerase as well as 3 beta-HSD is evidence that 2 alpha-[14C]BAP binds at the active sites of both enzyme activities. Because the same Arg-250 peptide has been affinity-alkylated in studies that targeted each of the two activities, we propose that the 3 beta-HSD and isomerase reactions are catalyzed in this region of the enzyme protein. PMID- 9419766 TI - Differentiation-dependent expression of the BCL-2 proto-oncogene in the human trophoblast lineage. AB - OBJECTIVE: We explored the role of the BCL-2 proto-oncogene in the life cycle of trophoblast cells by examining: 1) the patterns of BCL-2 expression in normal placenta at various gestational ages and in specimens of hydatidiform moles and choriocarcinomas, and 2) the effects of cyclic adenosine monophosphate (cAMP) treatment of JEG-3 choriocarcinoma cells, which induces differentiated functions, on BCL-2. METHODS: BCL-2 protein was localized by indirect immunofluorescence and immunoperoxidase staining of tissue sections and cells using monoclonal and polyclonal antibodies. Western and Northern blotting were used to assess BCL-2 and p53 protein and mRNA levels, respectively. JEG-3 cells were transfected with a BCL-2 expression plasmid to establish that BCL-2 protein could be expressed at high levels in this cell type. RESULTS: BCL-2 immunostaining was most prominent in the syncytiotrophoblast of normal placenta. It was found in syncytiotrophoblast of complete and partial hydatidiform moles, whereas cytotrophoblast staining was weak. BCL-2 immunostaining was also barely detectable in choriocarcinoma cells (JEG-3 cells) and a primary choriocarcinoma. However, BCL-2 protein could be transiently overexpressed in JEG-3 cells by transfection with an expression plasmid. Western blot analysis revealed low levels of BCL-2 in JEG-3 cells and a rise in BCL-2 protein in placental extracts from 10 weeks' gestation to term. In contrast, p53 protein was abundant in JEG-3 cells and normal placenta at 10 weeks' gestation, but low at term, BCL-2 transcripts were substantially more abundant in term placenta than in JEG-3 cells. Treatment of JEG-3 cells with 8-Br-cAMP, which induces genes characteristic of the syncytiotrophoblast, raised BCL-2 protein approximately twofold, whereas p53 mRNA declined. CONCLUSIONS: We conclude that: 1) There is a differentiation-dependent pattern of BCL-2 expression in the placenta, with the protein being most abundant in terminally differentiated trophoblast cells; 2) there appears to be an inverse relation between BCL-2 and p53 expression in trophoblast; and 3) cAMP regulates BCL-2 protein in trophoblast cells. We speculate that the expression of BCL-2 in terminally differentiated trophoblast cells, and hence resistance to apoptotic cell death, may be one mechanism by which trophoblast mass is preserved during pregnancy. Conversely, the relatively low expression of BCL-2 in choriocarcinoma cells may render them more susceptible to apoptosis. PMID- 9419767 TI - Cathepsin D activity in human ovarian tissues and ovarian carcinoma. AB - OBJECTIVE: This study determined levels of cathepsin D activity in tissue components of normal human ovary to establish a basis for comparison with human ovarian adenocarcinomas. METHODS: Cathepsin D activity per mg tissue, per microgram protein, and per microgram DNA was determined in human ovarian tissues (cortex, follicle, corpus luteum, corpus albicans) from patients of various ages and during the menstrual cycle. Levels of cathepsin D activity were also determined in ovarian adenocarcinomas and other pathologic tissues. RESULTS: Cathepsin D levels (per mg tissue) were significantly greater (P < .001) in ovarian follicle and corpus luteum compared with cortex. Although there was not a clear correlation between enzyme activity in the cortex and day of the menstrual cycle or patient age, levels of enzyme activity appeared to decrease with each parameter. Cathepsin D levels per mg tissue in ovarian adenocarcinoma were 40% higher than in postmenopausal ovarian cortex, but the difference was not statistically significant. CONCLUSION: The diversity of cathepsin D levels in normal ovarian tissue compartments indicates that specific tissues must be used in comparisons with ovarian tumors. PMID- 9419768 TI - Academic gynecologic oncology: where are we? PMID- 9419769 TI - Pathogenesis of ovarian cancers. AB - OBJECTIVE: To review our current understanding of the molecular genetic events involved in the development of epithelial ovarian cancers. METHODS: Molecular biologic techniques have been used to examine the role of growth-stimulatory genes (oncogenes) and -inhibitory genes (tumor suppressors) in ovarian cancer. RESULTS: A number of different peptide growth factors and their receptors are expressed by normal and malignant ovarian epithelial cells. However, the role, if any, of growth factors in ovarian carcinogenesis or maintenance of the transformed phenotype remains unknown. Amplification and overexpression of the HER-2/neu and c-myc oncogenes occur in a significant fraction of epithelial ovarian cancers (20-30%). Overexpression of HER-2/neu has correlated with poor survival in some studies, whereas c-myc amplification is more common in serous cancers. Mutation of the K-ras oncogene frequently occurs in borderline ovarian tumors, but is less common in invasive epithelial ovarian cancers. Mutation of the p53 tumor suppressor gene occurs in approximately half of advanced (stage III/IV) ovarian cancers and in 15% of early (stage IA/IB) cases. Most recently, preliminary studies have focused on the role of other tumor suppressor genes, cyclins, WAF1, and DNA mismatch repair genes. CONCLUSIONS: An understanding of the molecular events involved in the pathogenesis of epithelial ovarian cancer is beginning to evolve. Improvements in early diagnosis, treatment, and prevention of this deadly disease are dependent on further progress in this area. PMID- 9419770 TI - Changes in oxytocin receptor messenger RNA in the endometrium, myometrium, mesometrium, and cervix of sheep in late gestation and during spontaneous and cortisol-induced labor. AB - OBJECTIVE: Changes in oxytocin binding in intrauterine tissues have been demonstrated in relation to labor and delivery in several species using ligand binding techniques. Little information is available in any species on changes in mRNA for the oxytocin receptor in intrauterine tissues in relation to the changes in myometrial activity at term. The objective of this study was to quantify oxytocin receptor mRNA in critical intrauterine tissues in the pregnant sheep in relation to the myometrial electromyographic activity patterns that accompany labor. METHODS: Uterine tissues were removed under halothane general anesthesia from control ewes not in labor at two stages of gestation, 131 and 140-145 days, and from ewes in spontaneous term labor at 140-145 days' gestation. Tissues were also obtained from ewes in labor following the infusion of cortisol to the fetus beginning at 127 days' gestation. RESULTS: In both the myometrium and endometrium, oxytocin receptor mRNA was significantly increased in both spontaneous term labor and cortisol-induced labor as compared with appropriate gestational age-matched controls. In contrast, oxytocin receptor message was unchanged at the time of labor in the mesometrium and cervix in all groups studied. CONCLUSIONS: In the pregnant sheep, myometrial and endometrial oxytocin receptor mRNA increase significantly in both spontaneous and cortisol-induced labor as compared with appropriate controls. In contrast, there was no increase in oxytocin receptor mRNA in either the cervix or the mesometrium. PMID- 9419771 TI - Low serum estradiol and high serum progesterone concentrations characterize hypertensive pregnancies at high altitude. AB - OBJECTIVE: Intrauterine growth retardation and preeclampsia are more common at high than at low altitude. Because altered hormonal profiles have been linked with these disorders, we asked whether placental steroid hormone concentrations were altered during pregnancy at high altitude. METHODS: We measured progesterone, unconjugated estradiol, and estriol (by radioimmunoassay) at weeks 20, 30, and 36 of pregnancy in 18 women at low altitude (1600 m) and 40 women at high altitude (3100 m). RESULTS: Women at 3100 m compared with 1600 m had lower serum estradiol concentrations at week 36 of pregnancy, and lower estriol and higher progesterone concentrations throughout pregnancy. As a result, the progesterone/estradiol ratio was greater in the high- versus the low-altitude women. Estradiol fell between weeks 30 and 36 in women who developed transient hypertension or preeclampsia. The fall in estradiol was accompanied by a marked increase in progesterone concentrations among the preeclamptic women. At 3100 m, estradiol correlated negatively (r = -0.37, P < .05) and progesterone positively (r = 0.46, P < .05) with mean arterial pressure at week 36 of pregnancy. CONCLUSIONS: We speculate that reduced placental oxygen pressure (PO2) at high altitude may decrease placental aromatase activity and thereby lower estradiol and estriol concentrations. The factor(s) responsible for the rise in progesterone is unknown. Possibly, high progesterone relative to estradiol concentrations contributes to the development of preeclampsia at high altitude. PMID- 9419772 TI - Neonatal salvage by week's gestation in pregnancies complicated by HELLP syndrome. AB - OBJECTIVE: For clinical management and counseling purposes, we determined the neonatal salvage by weeks' gestation and birth weight of pregnancies complicated by HELLP syndrome (hemolysis, elevated liver enzymes, and low platelets). METHODS: All patients who delivered with a diagnosis of HELLP syndrome between January 1, 1980 and December 31, 1991 at a single tertiary care medical center were evaluated for neonatal outcome and survival. The syndrome was diagnosed in the presence of severe preeclampsia/eclampsia accompanied by laboratory evidence of hemolysis, hepatic dysfunction, and thrombocytopenia. RESULTS: During the study interval, 58,670 live-born deliveries occurred, of which 527 (0.11%) had HELLP syndrome. In this population, 143 patients delivered fetuses at less than 30 weeks' gestational age. Based upon gestational age intervals, neonatal salvage was 0% at 23 weeks in 13 deliveries, 17% (intact salvage 8.5%) at 24 weeks, 31% (intact salvage 15%) at 25 weeks, 75% (intact salvage 65%) at 26 weeks, 80% (intact salvage 70%) at 27 weeks, and 83% at 28 and 29 weeks (intact salvage 70 and 76%). Based on birth weight intervals, neonatal salvage was 0% at less than 600 g, 34% at 600-700 g (intact salvage 17%), 69% at 700-800 g (intact salvage 53%), and 84% or more at greater than 800 g. These pregnancy outcomes are similar to those in this institution in patients without HELLP syndrome. Disease severity was distributed evenly among the 143 patients at less than 30 weeks' gestation. Apart from gestational age, there was no significant relation between the severity of the HELLP disease process and ultimate neonatal salvage. CONCLUSIONS: Intact neonatal salvage in pregnancies complicated by HELLP syndrome is poor at weights less than 700 g and gestation of 25 weeks or less, but is more optimistic in pregnancies of greater than 700 g and 26 weeks' gestation or later. Aggressive efforts to enhance perinatal outcome, by operative delivery if indicated for fetal compromise, appear especially appropriate in gestations of greater than 700 g and 26 weeks' gestation or later, provided that adequate intensive care nursery facilities and neonatal expertise are available. PMID- 9419773 TI - Accelerated recovery from severe preeclampsia: uterine curettage versus nifedipine. AB - OBJECTIVE: We investigated the ability of uterine curettage and nifedipine to accelerate postpartum recovery from severe preeclampsia. METHODS: Forty-five parturients with severe preeclampsia were randomly assigned to one of three groups following delivery. Patients in group 1 were managed with intravenous magnesium sulfate (2 g/hour) and observed in the obstetric recovery room until blood pressure had stabilized (systolic blood pressure less than 150 mmHg and diastolic blood pressure less than 100 mmHg) and adequate diuresis was noted. Group 2 was treated in a similar manner but with the addition of oral nifedipine, 10 mg every 4 hours postpartum for 48 hours. Group 3 underwent an ultrasound directed curettage immediately following delivery in the delivery/operating room and was then treated as in group 1. All three groups were assessed postpartum for mean arterial pressure (MAP) and urine output (UO) every 2 hours, hematocrit and platelet count every 6 hours, and lactic dehydrogenase/aspartate aminotransferase every 12 hours for 48 hours postpartum. RESULTS: Fifteen women were assigned to each of the three treatment groups. The MAP decreased significantly (P < .0001) in all three groups during the first 48 hours postpartum. Treatment interaction indicated specific differences among the groups. Standard therapy (group 1) was significantly inferior to nifedipine (group 2) and curettage (group 3) in regard to MAP decrease (P = .0017) and UO increase (P = .0137). No statistical differences existed between nifedipine and curettage. The rise in the platelet count following delivery was significantly different among the three groups (P = .033), with a much more rapid recovery in the curettage group (12-18 hours) than in the other groups (P = .0106). CONCLUSIONS: Nifedipine and uterine curettage both appear to accelerate recovery from severe preeclampsia, as measured by MAP and UO. Uterine curettage appears the most effective in rapidly resolving the thrombocytopenia associated with severe preeclampsia. PMID- 9419774 TI - Enhancement of in vitro murine embryo development by recombinant leukemia inhibitory factor. AB - OBJECTIVE: Human recombinant leukemia inhibitory factor (rLIF) has been shown to stimulate hatching of murine and ovine embryos in vitro. The temporal and dose dependent effects of murine rLIF (mrLIF) and human rLIF (hrLIF) on embryo development in two different mouse strains were investigated in this work. METHODS: Two-cell embryos were recovered from the fallopian tubes of superovulated/mated females and cultured in Krebs medium plus bovine serum albumin in microdroplets under oil. RESULTS: In the B6CBF1 strain, mrLIF significantly stimulated blastocyst formation and decreased embryo fragmentation/degeneration when added simultaneously at the initiation of culture or 24 hours thereafter. Human rLIF also had a positive effect on development. In the CD1 strain (lower fecundity), mrLIF dose-dependent effects were observed, with enhanced developmental stimulation achieved with higher doses. CONCLUSIONS: These findings confirm that hrLIF stimulates mouse embryo development in vitro and that different mouse strains show distinct responses to the cytokine. In addition, mrLIF enhances blastocyst formation and decreases embryo fragmentation when added to the embryo culture as early as the two-cell stage. PMID- 9419775 TI - Identification of cyclo(His-Pro)-like immunoreactivity in human follicular fluid: correlation with steroid and peptide hormones. AB - OBJECTIVE: The purpose of this study was to evaluate human follicular fluid (FF) for the presence of cyclo(His-Pro)-like immunoreactivity (CHP-LI). After verifying its presence, we quantitated the levels and investigated correlations with other follicular parameters, including hormone levels. METHODS: Follicular fluid was collected from female volunteers undergoing controlled ovarian hyperstimulation. Fluid was collected by follicular puncture, either transvaginally (in vitro fertilization) or laparoscopically (gamete intrafallopian transfer) at the time of oocyte retrieval (N = 137). Follicular size was determined ultrasonographically. Assays for steroid and peptide hormones were determined with commercially available radioimmunoassay kits. CHP-LI was measured using a previously reported assay; parallel dilution curves and column chromatography aided in immunoidentity. RESULTS: The mean FF CHP-LI concentration (13.10 +/- 1.83 nmol/L, N = 137) was greater than the corresponding serum values (9.42 +/- 2.45 nmol/L; N = 21) (P < .05). Large follicles (20 mm or greater; 14.45 +/- 1.74 nmol/L) contained significantly more CHP-LI than either medium follicles (16-19 mm; 11.51 +/- 1.88 nmol/L) or small follicles (15 mm or smaller; 10.83 +/- 2.12 nmol/L) (P < .05). Positive correlations were found between FF CHP LI values and corresponding FF progesterone and prolactin concentrations (r = 0.67 and 0.62, respectively; P < .05). CONCLUSION: Mean CHP-LI levels in the FF are greater than those in the corresponding serum. We suggest that the neuropeptide may be originating from either peptidase cleavage of precursor peptides or from granulosa cell production. PMID- 9419776 TI - Characterization of paracellular permeability in cultured human cervical epithelium: regulation by extracellular adenosine triphosphate. AB - OBJECTIVE: The purpose of the present study was to compare the permeability and regulation of paracellular transport in human cervical cells with those in epithelial cells of other organs. METHODS: Cervical cells (ECE16-1, Caski, and HT3) were grown on filters, and transepithelial electrical conductance (GT) and the permeability to pyranine (PPyr) were determined. RESULTS: Cervical cultures were characterized by high GT (83-125 mS.cm-2) and high PPyr (6.2-18 x 10(-6).sec 1). The GT was not significantly affected by cell density but was increased by 20% by lowering extracellular calcium to 0.45 mmol/L or less. The high values of GT and PPyr and the regulation by extracellular calcium indicate that all three cervical cell lines have "leaky" tight junctional complexes. Addition of extracellular adenosine triphosphate (ATP) at 50 mumol/L to the cervical cultures evoked a biphasic change in GT that was unique to the cervical cells: an initial increase, followed by a sustained decrease by 30% from baseline GT. The decrease of GT was associated with a decrease in PPyr by 17%, indicating that ATP had an effect on the tight junctional/paracellular permeability. The ATP effect was reversible either by washing or by chemical hydrolysis with ATPase. The non cervical cell lines all responded to extracellular ATP with a transient increase in GT, but not with the pronounced decrease. CONCLUSION: The permeability of the paracellular pathway can be regulated in cervical epithelia by mechanisms that may be different from those in epithelial cells from other organs. PMID- 9419777 TI - Identification of p53 mutations in endometrial adenocarcinoma by polymerase chain reaction-single-strand conformation polymorphism. AB - OBJECTIVE: We determined whether mutations in p53 exons 5-6-7-8, as detected in the polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) test, might be correlated with stage or grade in endometrial adenocarcinoma. METHODS: We amplified sequences containing exons 5, 6, 7, or 8 in DNA from tumors and controls. Mutation within the amplified sequences was indicated by changes in electrophoretic mobility (band shifts) in the SSCP test. The results were analyzed statistically and compared with the results of other, similar studies. RESULTS: We identified 15 band shifts among 47 endometrial tumors studied (band shifts in two different exons in two cases) and none among 42 controls. Band shifts in exons 5 and 8 were associated uniformly with grade 2 or grade 3 histology. In other studies p53 mutations were correlated with advanced-stage malignancy. CONCLUSION: Further evaluation of particular p53 mutations and their relation to disease course in endometrial adenocarcinoma seems warranted. The PCR SSCP test seems well-suited to this purpose. PMID- 9419779 TI - Providing a "Noah's Ark" for research in the reproductive sciences by fostering young investigators: a role for our journal. PMID- 9419780 TI - Strategies of ovarian function of importance to gynecologic investigations. AB - OBJECTIVE: The purpose of this article was to examine the similarities and differences in ovarian morphology and function that are known to exist among mammalian species. The previous difficulties and present solutions to the characterization of ovarian function in non-domestic or non-laboratory species are discussed. METHODS: The current literature regarding comparative ovarian structure and function was reviewed to identify examples of the diversity found with mammals. RESULTS: Major differences in ovarian anatomy include location in the body cavity; the degree of membranous covering surrounding the ovary; vascularization of the ovary, oviduct, and uterus; and the organization of different cell types within the ovary. The differences in ovarian function are simplified in this review by categorizing the known examples into six groups according to patterns in follicle growth and development, mechanisms of ovulation, response of steroid target cells, and luteal function. CONCLUSIONS: Each mammalian species has a unique expression of its ovarian function. The differences between species range from quantitative differences in ovarian cycle phase length to qualitative differences in cell type, endocrine activity, and mechanism of ovulation. These differences present a special problem to research scientists when they attempt to select models for studying many aspects of female reproduction. PMID- 9419778 TI - Estrogen receptors are identified in the glioblastoma cell line U138MG. AB - OBJECTIVE: The antiestrogen tamoxifen has been found to be effective in decreasing glioblastoma cell proliferation, but the mechanism underlying this effect and whether it is through the estrogen receptor (ER) is controversial. The objective of this study was to determine whether ERs are present in three human glioblastoma cell lines--HS683, U138MG, and JHN J889H--using the most sensitive techniques available. METHODS: Ligand binding and flow cytometry were employed to identify estrogen and progesterone receptors. The reverse transcriptase polymerase chain reaction was used to identify ER mRNA, and a novel reporter gene transfection assay demonstrated that the ER was capable of activating gene transcription. RESULTS: U138MG glioblastoma cells contain ERs that are capable of increasing gene transcription in response to estradiol. No ERs were found in HS683 or JHN J889H cells. CONCLUSION: Tamoxifen may be acting through the ER in some glioblastoma cells. PMID- 9419781 TI - Regulatory effects of multifunctional cytokines and steroid hormones on apolipoprotein B production by human fetal hepatocytes. AB - OBJECTIVE: The purpose of the present investigation was to determine the effects of certain multifunctional cytokines (tumor necrosis factor-alpha [TNF-alpha], interleukin-1 beta [IL-1 beta], and IL-6) and steroid hormones (estradiol, testosterone, progesterone, and cortisol) on the production of apolipoprotein B (Apo B) by cultured human fetal hepatocytes. We conducted these experiments because of our recent observations that purified human fetal Kupffer cells produce TNF-alpha, IL-1 beta, and IL-6. METHODS: Human fetal hepatocytes, specifically depleted of hematopoietic precursors and Kupffer cells, were cultured in defined medium. The amounts of Apo B released into the culture medium were measured by a sensitive and specific enzyme-linked immunosorbent assay. RESULTS: Hepatocytes (10(6) cells) produced 82 +/- 16 ng Apo B per 24 hours during days 4-7 in culture. Results demonstrated that treatment of cultured hepatocytes with TNF-alpha maximally inhibited Apo B production by 50% at a half maximal concentration of 100 pg/mL, whereas IL-1 beta maximally inhibited Apo B production by 80% at a half-maximal dose of 200 pg/mL. Cells exposed to IL-6 produced increased amounts of Apo B, but only after IL-6 was removed from the culture medium. The addition of TNF-alpha, IL-1 beta, or IL-6 did not significantly affect hepatocyte viability. At physiologic concentrations (1 mumol/L), estrogens were able to increase the production of Apo B by 25-65%; however, no positive or negative effect could be demonstrated with dexamethasone, cortisol, testosterone, or progesterone. When using synthetic estrogens such as ethinyl estradiol and mestranol, the stimulatory effect was most pronounced. CONCLUSION: Tumor necrosis factor-alpha and IL-1 beta have an inhibitory effect and estrogens have a stimulatory effect on Apo B production by human fetal hepatocytes in culture. These studies suggest that fetal hepatocytes can produce Apo B and that the synthesis of Apo B is under the control of multifunctional cytokines and steroid hormones. PMID- 9419782 TI - Amniotic fluid interleukin-6 and interleukin-8 levels predict the success of tocolysis in patients with preterm labor. AB - OBJECTIVE: To determine whether the levels of the cytokines interleukin-6 (IL-6) and IL-8 in amniotic fluid identify patients with preterm labor who are resistant to tocolysis. METHODS: Amniocenteses were performed in 23 women with documented preterm labor at 20-32 weeks' gestational age who were treated subsequently with tocolytics. The concentrations of IL-6 and IL-8 in amniotic fluid were determined by double-antibody radioimmunoassay methods using recombinant human IL standards. RESULTS: Of the 23 patients, five failed to respond to tocolysis (four delivered within 48 hours), and of the remaining 18, all delivered more than 9 days after tocolysis was initiated (mean 31 +/- 10 days; range 9-61). In women who had failed tocolysis, discriminatory concentrations of IL-6 and IL-8 were 20 and 15 ng/mL, respectively. Of the patients who had amniotic fluid concentrations higher than these thresholds, all failed tocolysis (100% positive predictive value) and delivered within 6 days. The patients with levels below these discriminatory concentrations had successful tocolysis, and 17 of 18 delivered more than 2 weeks after treatment (95% negative predictive value). CONCLUSION: The success of tocolysis and thus delivery remote from an episode of preterm labor is associated with discriminatory amniotic fluid IL-6 and IL-8 levels of less than 20 and less than 15 ng/mL, respectively. If the immunologic response that causes the release of IL-6 and IL-8 has not occurred, the likelihood of successful tocolysis is extremely high. However, if both IL-6 and IL-8 are elevated, tocolysis is likely to fail and delivery may occur within 48 hours. PMID- 9419783 TI - Epidermal growth factor, transforming growth factor-alpha, and epidermal growth factor receptor localization in the baboon (Papio anubis) oviduct during steroid treatment and the menstrual cycle. AB - OBJECTIVES: Polypeptide growth factors may modulate the actions of estrogen (E2) and progesterone (P) in reproductive tissues in an autocrine/paracrine manner. The objective of this study was to determine whether the baboon oviduct contains epidermal growth factor (EGF), transforming growth factor-alpha (TGF alpha), and EGF receptor (EGF-R) and whether changes in their expression are correlated with various hormonal states. METHODS: Oviductal tissue was obtained from adult female baboons (Papio anubis) after oophorectomy and steroid treatment, and during the menstrual cycle. Ampullary regions were fixed in Bouin's fixative and embedded in paraffin for immunocytochemistry using rabbit polyclonal antibodies against EGF and EGF-R, and mouse monoclonal antibody against TGF alpha. RESULTS: Both EGF and EGF-R were present in all tissue compartments (most strongly in the epithelium, followed by smooth muscle and stroma) at all reproductive stages and showed similar staining patterns. However, the most intense immunoreactive product was found in the tissue obtained from the E2-treated and late follicular phase animals. At this time, intense staining was present in the apical regions of the mature ciliated cells, whereas the stain was dispersed uniformly over the cytoplasm of all other cell types. Immunoreactive TGF alpha was limited primarily to the nonciliated epithelial cells, and staining was most intense in the E2 treated and late follicular phase tissues. Transforming growth factor-alpha formed intense perinuclear deposits in the mature secretory cells, an area that corresponds to the Golgi region. No immunoreactive product was observed for any of these proteins when preimmune serum was substituted for the primary antibody or when the primary antibody was preabsorbed with antigen. CONCLUSION: In summary, EGF, TGF alpha, and EGF-R are present in the ampulla of the baboon oviduct. Moreover, the localization and intensity of immunoreactive product are dependent on cell type and hormonal state. These data are consistent with the concept that EGF, TGF alpha, and EGF-R may be regulated by E2 and P and thus may play a role in cell differentiation and function. In addition, the specific localization of TGF alpha suggests that this growth factor may be synthesized for release from the secretory cells and thus may also function as a modulator of gamete/embryo viability and development. PMID- 9419784 TI - Epidermal growth factor, transforming growth factor-alpha, and epidermal growth factor receptor localization in the baboon (Papio anubis) uterus during the menstrual cycle and early pregnancy. AB - OBJECTIVE: Marked alterations occur in the synthesis of endometrium-specific proteins during the first third of pregnancy in the baboon. Because epidermal growth factor (EGF) expression has been associated with proliferation in the human and mouse endometrium, we hypothesized that EGF, transforming growth factor alpha (TGF alpha), and EGF receptor (EGF-R) expression in baboon endometrium may be modulated by the early invasive trophoblast and play a role in decidualization of the endometrial stroma. METHODS: Endometrial tissue was obtained from cycling baboons (n = 4-5 per time point), ovariectomized steroid-treated baboons (n = 4 per group), or from pregnant baboons on days 18-60 of pregnancy (n = 2-4 per group). The tissue was fixed in Bouin's solution and embedded in paraffin for immunocytochemistry using polyclonal antibodies against EGF and EGF-R and a monoclonal antibody to TGF alpha. RESULTS: Endometrial staining was located almost entirely in the glandular epithelium for TGF alpha and EGF-R in the follicular phase animals, whereas EGF staining was strongest in the periglandular stroma. In the luteal phase, specific staining for EGF also was detected in the glands as well as the periglandular stroma. There appeared to be little difference in endometrial staining between the late follicular and mid-luteal phase for TGF alpha and EGF-R. A similar pattern was observed in the steroid treated animals. In the endometrium from pregnant animals, EGF, TGF alpha, and EGF-R intensely stained the glandular epithelium on days 18, 25, and 32. Both EGF and EGF-R showed light stromal staining on days 18 and 25. Light stromal TGF alpha staining was present on day 25 and became moderately intense by day 32. By day 60, the most intense staining for EGF and EGF-R was stromal. Staining of TGF alpha continued to be strong in the remaining epithelium through day 60. In placenta, EGF and EGF-R intensely stained the syncytiotrophoblast, but not the cytotrophoblast, whereas TGF alpha stained only the villous cytotrophoblast and intermediate cytotrophoblast within maternal blood vessels. There appeared to be no change in this staining pattern or intensity in the placenta throughout early pregnancy. CONCLUSIONS: This study demonstrates the presence of EGF, TGF alpha, and EGF-R in the endometrium during the cycle and early pregnancy. The detection of EGF, TGF alpha, and EGF-R in the stromal cells during pregnancy correlated with the onset of decidualization. We propose that EGF, TGF alpha, and EGF-R may play a role in glandular development during the cycle and in decidualization and implantation during early pregnancy. PMID- 9419785 TI - Growth hormone receptor gene expression in the mouse uterus: modulation by gonadal steroids. AB - OBJECTIVES: We hypothesized that the mouse uterus expresses a growth hormone receptor (GHR) and that mouse endometrial GHR mRNA may undergo in vivo regulation by estradiol (E2) and progesterone (P). METHODS: Two weeks after bilateral ovariectomy, 35-day-old Balb/c mice were randomly assigned to receive either vehicle injections; E2 priming (300 ng/day) on days 1-6 and E2 (50 ng/day) on day 11; P (1 mg/day) on day 8-11; or E2 priming with P (days 8-11) and E2 on day 11, with or without RU 486 on day 8-11. At sacrifice 24 hours after the last injection, the uteri were processed for RNA extraction and in situ hybridization for GHR mRNA. Reverse transcriptase polymerase chain reaction amplification of uterine mRNA was performed to establish whether treatment with gonadal steroids differentially affected GHR gene expression. The GHR primers flanked a 326-bp region from the intracellular domain sharing no homology to the prolactin receptor or to the GH-binding protein. A 48-bp digoxigenin-labeled oligoprobe was used for in situ hybridization. RESULTS: Densitometry analysis of polymerase chain reaction products from total uterine mRNA revealed similar GHR mRNA expression in vehicle- and estrogen-treated animals. The addition of progesterone reduced GHR mRNA expression. In situ hybridization localized the GHR mRNA to the endometrium, glands, stroma, and myometrium. Stromal staining was reduced in the progesterone-treated mice. CONCLUSIONS: The identification of GHR message suggests that the mouse uterus is a site of action of GH. The lack of modulation of epithelial GHR by sex steroids does not support a role of GHR in this compartment. PMID- 9419786 TI - In vivo effects of a potent GnRH antagonist ORG 30850: physiologic evidence that down-regulation of GnRH receptors does not occur. AB - OBJECTIVE: Our purpose was to determine the pituitary responsiveness to exogenous GnRH in GnRH antagonist-suppressed ovariectomized monkeys. METHODS: This was a prospective experimental non-human primate study performed at the research laboratories of The Jones Institute for Reproductive Medicine. Seventeen long term ovariectomized cynomolgus monkeys were studied. INTERVENTIONS: The GnRH antagonist ORG 30850 was administered to long-term ovariectomized monkeys assigned to one of six groups: single subcutaneous injections in group A (n = 4), 0.3 mg/kg; group B (n = 4), 1.0 mg/kg; and group C (n = 3), 3.0 mg/kg; and six consecutive daily subcutaneous injections in group D (n = 2), 0.3 mg/kg; group E (n = 2), 1.0 mg/kg; and group F (n = 2), 3.0 mg/kg. Blood samples were collected daily from 10 days before treatment until 22 days after treatment, then weekly for 6 additional weeks. Intravenous GnRH stimulation tests (10 micrograms/kg) were performed on the day after vehicle injection (control) and the day after completion of treatment(s), and then at weekly intervals. The main outcome measures were serum levels of LH, FSH, and ORG 30850. RESULTS: Administration of ORG 30850 resulted in suppression (P < .05) of LH and FSH in all treatment groups. Long-term suppression (greater than 2 weeks) was evident in all primates receiving a cumulative dose of at least 1 mg/kg. Paradoxically, the responsiveness of the pituitary to exogenous GnRH was accentuated during the time of maximal tonic LH/FSH suppression. CONCLUSIONS: ORG 30850 is a potent long acting GnRH antagonist. Furthermore, the present in vivo demonstration of heightened pituitary responsiveness to exogenous GnRH emphasizes the divergent mechanisms of action of GnRH antagonists and GnRH agonists. PMID- 9419787 TI - Detection of apoptosis in human and rat ovarian follicles. AB - OBJECTIVE: The purpose of this study was to determine whether cells acquired from individual human preovulatory follicles undergo apoptosis (physiologic cell death) and, if so, to correlate the degree of apoptosis with characteristics of the follicles or the oocytes derived from the follicles. METHODS: We devised a sensitive nonradioactive method for detecting apoptotic DNA fragmentation in small numbers of cells derived from rat atretic follicles and follicular aspirates of patients undergoing assisted reproductive technologies. RESULTS: Using this method, apoptotic DNA was detected in rat atretic follicles, with optimal detection at 10-100 ng. Furthermore, apoptotic DNA was detected in some, but not all individual human follicular aspirates from several patients, and was found in follicles that produced oocytes that fertilized and developed into embryos. CONCLUSION: Apoptosis occurs in cells from human ovarian preovulatory follicles and may be a normal physiologic process of the follicle during luteinization. PMID- 9419788 TI - The anabolic effects of progesterone on fetal rat calvaria in tissue culture. AB - OBJECTIVE: We examined the effects of progesterone and cortisol on [3H] thymidine incorporation into DNA, total DNA content, and [3H] proline incorporation into collagen using a bone organ culture system. METHODS: The study consisted of nine experiments conducted at the University of Connecticut after approval by the animal care committee. Fetal rat half calvaria were cultured in the presence of progesterone (10(-5) to 10(-15) mol/L) and/or cortisol (10(-7) mol/L). Statistical analysis was carried out by analysis of variance. RESULTS: There was a biphasic increase in [3H] thymidine incorporation and DNA content of the tissue sample with progesterone treatment, with peaks at both high and low concentrations. Significant increases in [3H] proline incorporation were noted at concentrations of 10(-6) to 10(-10) mol/L of progesterone. Similar responses were observed in the presence of cortisol. CONCLUSIONS: Progesterone increased [3H] thymidine incorporation into DNA, total DNA content, and [3H] proline incorporation into collagen proteins in cultured fetal rat calvaria over a wide range of progesterone concentrations in the presence and absence of cortisol. PMID- 9419789 TI - Messenger RNA decay of macrophage colony-stimulating factor in human ovarian carcinomas in vitro. AB - OBJECTIVE: Recently, the importance of the macrophage colony-stimulating factor (CSF-1) and its receptor (encoded by the c-fms proto-oncogene) in patients with epithelial ovarian cancer has been recognized. Overexpression of CSF-1 denotes poor prognosis. Macrophage colony-stimulating factor may be one of a group of inflammatory cytokines, whose 3' untranslated region (UTR) contains AU-rich stretches and whose expression may be largely dependent on mRNA decay. The purposes of this study were to investigate the effect of protein synthesis inhibition on CSF-1 transcript expression, to help determine whether there is a role for labile intermediary proteins in the regulation of CSF-1 expression, and to explore the transcriptional or post-transcriptional mechanisms that could underlie such overexpression of CSF-1 transcripts by protein synthesis inhibitors. Although regulation of CSF-1 gene expression has been investigated in hematopoietic cells, such studies have not been carried out in any epithelial cancer. METHODS: Northern blot analyses for CSF-1 expression were performed on total cellular RNA extracted from primary and established ovarian cancer cell lines in the absence or presence of proteins synthesis inhibitors with different modes of action. The probe for the AU-rich exon 10 of CSF-1 was prepared by amplification of the terminal 143 bp of the 3' UTR of human CSF-1 by polymerase chain reaction. Transcription rates were assessed in the presence or absence of cycloheximide (CH) in nuclei of ovarian cancer cells by run-off analyses. The effect of CH on CSF-1 mRNA half-life was measured by actinomycin D chase experiments in SKOV3 cells. RESULTS: We demonstrate that CSF-1 mRNA was expressed by all of a panel of primary and established ovarian cancer cell lines. There are at least three CSF-1 transcripts expressed by ovarian cancer cells; we demonstrate that the 4.2-kb CSF-1 transcript contains exon-6 sequences, which are spliced from the 3.4- and 1.9-kb transcripts, whereas both the 4.2- and 3.4-kb CSF-1 transcripts contain the 3' AU-rich exon 10. Treatment with several different protein synthesis inhibitors resulted in marked overexpression of CSF-1 transcript levels, suggesting a potential role for labile proteins in the regulation of CSF-1 expression. The predominant effect of CH is on the two CSF-1 transcripts that contain exon 10. Although CH does not change the rate of CSF-1 gene transcription, measurement of CSF-1 mRNA stability reveals a prolongation of CSF-1 transcript half-life by CH from 4.5 hours to significantly greater than 6 hours in ovarian cancer cells. CONCLUSIONS: Augmented CSF-1 transcript stability underlies the marked overexpression of CSF-1 seen with protein synthesis inhibitors. Our results suggest the involvement of a labile regulatory protein that contributes to CSF-1 mRNA decay in ovarian cancer cells. Our data suggest further that exon 10 (not the spliced exon-6 sequences) of the CSF-1 transcript may contain an instability determinant. PMID- 9419791 TI - Energy medicine and the unifying concept of information. AB - Alternative medicine remains alternative because it poses serious challenges to the mainstream biomedical paradigm of mechanical reductionism and because it requires a new framework. This paper explores some of the hypotheses and challenges of energy medicine including healer interventions, electromagnetic therapies, and homeopathy. Together with new findings from the bioelectromagnetic field, they spell out the rudiments of a new paradigm for biology and medicine based on information. Information embraces the complex network of relations in the matter and energy transactions of living systems. It offers a unified view of energy medicine modalities as well as a fresh perspective for biology and medicine and new questions for further research. PMID- 9419790 TI - A cross-cultural comparison of four healing models. AB - Although Western medicine has tended to ignore other systems of preventing and treating disease and illness, practitioners of these systems serve more of the world's population than do allopathic practitioners. This paper contrasts allopathy with Chinese medicine, curanderismo, and a Native American healing system, using a 12-facet model. It points out the importance of knowing about alternative models of healing, especially in regard to religious and spiritual problems, in a multicultural society. PMID- 9419792 TI - Larry LeShan: mobilizing the life force, treating the individual. Interview by Bonnie Horrigan. PMID- 9419793 TI - Treating the cause--without causing the problem. PMID- 9419794 TI - Roots of healing: the new medicine. PMID- 9419795 TI - Summary of a report by the Health Council of The Netherlands on alternative modes of treatment under scientific investigation. PMID- 9419796 TI - Christiane Northrup, MD: medical practice as a spiritual journey. Interview by Bonnie Horrigan. PMID- 9419797 TI - Health aging mandates integration of conventional and alternative/complementary medical interventions. PMID- 9419798 TI - $115 million in breast cancer research funds available; alternative projects encouraged. PMID- 9419799 TI - An introduction to Ayurveda. AB - Ayurveda is a Sanskrit word derived from two roots: ayur, which means life, and veda, knowledge. Knowledge arranged systematically with logic becomes science. During the due course of time, Ayurveda became the science of life. It has its root in ancient vedic literature and encompasses our entire life, the body, mind, and spirit. PMID- 9419800 TI - Candace Pert, PhD: neuropeptides, AIDS, and the science of mind-body healing. Interview by Bonnie Horrigan. PMID- 9419802 TI - The mechanism of homeopathy? All that matters is that it works. PMID- 9419801 TI - OAM funding: a shared responsibility. PMID- 9419803 TI - The internal mystery plays: the role and physiology of the visual system in contemplative practices. AB - In summary, the "White Light" and the imagery reported in contemplative practices appears to be a complex system of reflexes that mediate ischemia to the cerebral cortex, stimulating "release" of the occipital poles and the rostral midbrain to discharge these images in any profound state of cellular agony. These vascular events are mediated by way of the autonomic nervous system, either accidentally or purposefully, thus acquiring a near-death experience. Understanding the mechanism of the reflexes has taken many years and thousands of researchers to elucidate. In spite of these discoveries and those yet to come, no one will ever fully know why these reflexes and the sensations associated with them exist, or why they cause such profound psychic, physical, and spiritual changes in the individuals who have them. At this point, I can only be humbled by the process that has brought me to the paradigm I have described. I only know that most of the connections and correlations expressed in this article were the product of some of those methods mentioned herein; therefore, I cannot, and will not, take credit for that portion provided by the Guiding Spirit. Thus, the internal mysteries of the brain will always hide the Inner Mysteries, although the heart will always have a way of finding the right answers. We have become the Uroboric serpent (the serpent that eats its tail), in that we have returned to the ancient Egyptians' way of considering the eye as the seat of the soul and the doorway into the Inner Mysteries of the religious experience. Now, we find t hat in many ways they were correct that the eye, through its physiology, is another key to understanding these Inner Mysteries. PMID- 9419804 TI - David Reilly, FRCP, MRCGP, FFHom: research, homeopathy, and therapeutic consultation. Interview by Bonnie Horrigan. PMID- 9419805 TI - The interface between traditional and alternative medicine. PMID- 9419806 TI - Excavation of the 1814 Battle of Snake Hill site: a medical history perspective. PMID- 9419807 TI - Human remains: issues confronting museums and the scholarly disciplines. PMID- 9419808 TI - When the patient is Abraham Lincoln. PMID- 9419809 TI - Advisory statement by the panel on DNA testing of Abraham Lincoln's tissue. PMID- 9419810 TI - A symposium on Sigmund Freud and art at the David and Alfred Smart Museum of Art. PMID- 9419811 TI - Transglutaminase II: a new class of GTP-binding protein with new biological functions. AB - Tissue type transglutaminase (TGase II) is historically a member of the transglutaminase family, which covalently cross-links cellular proteins and polyamines. A recent new finding in the TGase II field is that the enzyme functions as a signal mediator from receptors to an effector in transmembrane signaling. This review will discuss the recent development of TGase II. This new signal transducer was termed Gh when initially discovered and was recently found to be TGase II. To help the reader understand the role of Gh as a signal mediator, the role of heterotrimeric G-proteins in hormone-mediated transmembrane signaling is briefly discussed. We have highlighted how Gh transmits the alpha 1 adrenoceptor signal to the phospholipase C-delta 1 and how Gh is activated and deactivated compared to the prototype of heterotrimeric G-proteins. Recent developments regarding the structure-function of Gh and other biological functions of Gh are discussed to facilitate understanding the impact of Gh in cells. PMID- 9419812 TI - Signalling mechanisms in erythropoiesis: the enigmatic role of calcium. AB - The glycoprotein hormone, erythropoietin is the principal regulator of the production of circulating erythrocytes by controlling proliferation, differentiation and survival of its target erythroid progenitor cells. The receptor for erythropoietin is a type I cytokine receptor lacking intrinsic tyrosine kinase activity. It mediates tyrosine phosphorylation through its association with nonreceptor tyrosine kinases such as JAK2 and initiates a cascade of signalling events in response to erythropoietin. Significant progress has been made in identifying signalling pathways triggered by erythropoietin. However, the exact signalling mechanisms mediating the known physiological effects of erythropoietin in erythroid progenitor cells are poorly understood. There are many open questions including the role of Ca2+ in erythropoietin induced signal transduction. Although the results concerning the effect of erythropoietin on [Ca2+]i in various erythroid cells are conflicting, [Ca2+]i increasing agents mimic the effect of erythropoietin on c-myb expression and activate the program of haemoglobin synthesis in murine erythroleukemia cells. An attempt is made in this review to survey recent data on the erythropoietin induced signal transduction with respect to the different physiological effects of this hormone. PMID- 9419813 TI - G-protein activation, intracellular Ca2+ mobilization and phosphorylation studies of membrane currents induced by AlF4- in Xenopus oocytes. AB - We have examined the electrophysiological responses induced by aluminium fluoride (AlF4-) and carbachol in Xenopus oocytes. Application of AlF4- induced Ca(2+) dependent oscillatory and smooth Cl- currents. Pre-treatment of oocytes with microinjected guanosine 5'-O-(2-thiodiphosphate) diminished the currents, indicating that the effect of AlF4- occurred through G-protein activation. Confocal imaging of intracellular Ca2+ clearly demonstrated that AlF4- could increase the internal Ca2+ concentration in oocytes in the absence of external Ca2+. A protein kinase (PK) activator (4-beta-phorbol 12,13-dibutyrate) decreased the AlF4(-)-induced membrane currents, whereas a PK inhibitor (staurosporine) caused an increase. On the other hand, the protein phosphatase inhibitor (okadaic acid) showed little effect. Although the effects of the phosphorylating/dephosphorylating agents on the carbachol-induced currents were qualitatively similar to the case of AlF4-, some quantitative differences was noted. The results are discussed in terms of the signaling pathways involving muscarinic receptors and G-protein(s) in Xenopus oocytes. PMID- 9419814 TI - Angiotensin II-induced inositol phosphate generation is mediated through tyrosine kinase pathways in cardiomyocytes. AB - The objective of this study was to determine whether the G-protein-linked angiotensin II receptor mediated inositol phosphate production involves a tyrosine phosphorylation (tyr phos) dependent pathway in the heart. Cardiomyocytes, in culture, from 7-day-old chick embryonic hearts were incubated with myo [3H] inositol for 18-24 h. Cells were incubated with LiCl to inhibit inositol 1-phosphate phosphatase and allow accumulation of inositol phosphates with angiotensin II (ang II) treatment. Inositol fractions were separated on column chromatography. Ang II produced significant (p < 0.01) increases of InsP1, InsP2, and InsP3, within 1 min of treatment of cardiomyocytes. Tyrosine kinase inhibition with genistein significantly (p < 0.05) reduced ang II induced inositol phosphate production. This did not occur with the analogue diazdien that is a very weak inhibitor of tyrosine kinase. The ability of ang II to induce tyr phos was demonstrated in whole cell lysates of cardiomyocytes immunoprecipitation with anti-P-Tyr antibodies. Genistein blunted this action of ang II. The rapid activation of a tyr phos dependent pathway by ang II was demonstrated by the similar time course of tyr phos of two different cardiac proteins, 70 and 195 kDa, and peak inositol phosphate production. Tyr phos of these cardiac proteins was mediated predominantly but not exclusively through the AT1 and II receptor subtype as it was completely blocked by the AT1 antagonist losartan, while the AT2 receptor antagonist PD123319 blunted ang II-induced tyr phos. These results demonstrate a novel role for a tyr phos dependent pathway in the heart for ang II induced inositol phosphate production and strengthens the concept of the interaction of G-protein coupled receptors with tyrosine kinases. PMID- 9419815 TI - Wortmannin interferes with thrombin-evoked secondary calcium redistribution in human platelets. AB - Wortmannin has previously been reported to inhibit calcium entry in thrombin stimulated human platelets. We extend these findings by demonstrating that the redistribution of calcium from intracellular stores features two separate, consecutive phases the second of which is selectively abolished by wortmannin. The primary release of calcium from Ins 1,4,5 P3-sensitive stores remains unaffected. Hence, wortmannin is interfering with regulation of any secondary, sustained calcium accumulation in the cytosolic compartment of activated platelets, originating either from intracellular stores or from calcium entry. We assume that wortmannin blocks a common step in receptor-dependent regulation of calcium entry. We assume that wortmannin blocks a common step in receptor dependent regulation of calcium entry and intracellular calcium circulation. PMID- 9419816 TI - Identification and characterisation of a human calmodulin-stimulated phosphodiesterase PDE1B1. AB - A cDNA encoding a calmodulin-stimulated 3',5'-cyclic nucleotide phosphodiesterase (PDE) was isolated from a human brain cDNA library. The cDNA, designated HSPDE1B1, encoded a protein of 536 amino acids that shared 96% sequence identity with the bovine "63 kDa" calmodulin-stimulated PDE. The recombinant protein had cyclic nucleotide phosphodiesterase activity that was stimulated approximately 2 fold by Ca2+/calmodulin and preferred cGMP as substrate. In addition, the enzymatic activity of HSPDE1B1 was inhibited by phosphodiesterase inhibitors with potencies similar to that displayed toward the bovine PDE1 enzymes: IBMX approximately equal to 8-methoxymethyl-IBMX > vinpocetine approximately equal to zaprinast > cilostamide > rolipram. HSPDE1B1 mRNA was found predominantly in the brain. Lower mRNA levels were found in heart and skeletal muscle. In situ hybridisation of brain revealed expression of HSPDE1B1 predominately in neuronal cells of the cerebellum, hippocampus and caudate. The HSPDE1B1 gene was mapped to human chromosome 12. A partial genomic sequence of HSPDE1B1 was isolated and shown to contain two splice junctions that are conserved in the rat PDE4 and the Drosophila dunce genes. PMID- 9419817 TI - Cyclic AMP-independent activation of neutrophil-like HL-60 cells by prostaglandin E2. AB - The role of cAMP in mediating prostaglandin E2 (PGE2)-stimulated aggregation of neutrophil-like HL-60 cells has been investigated. Although the EP2 receptors appear to couple to Gs-proteins, PGE2 stimulated HL-60 cell aggregation appears to be a cAMP-independent process. This response to PGE2 in independent of calcium and tyrosine kinase activity, appears to involve activation of phosphatidylinositol 3-kinase which is negatively regulated by phosphatidic acid generated from phospholipase D activity, and is partially dependent on protein kinase C activity. In contrast, although the chemotactic peptide N-formyl methionyl-leucyl-phenylalanine (FMLP) produces a similar aggregation response to PGE2, FMLP uses a distinct intracellular signalling pathway. The aggregation response to FMLP involves activation of Gi-proteins, is partially dependent on extracellular calcium, is negatively regulated by protein kinase C, and is independent of phosphatidylinositol 3-kinase, phospholipase D and tyrosine kinase activity. The possibility exists that EP2 receptor activation leads to Gs dependent, but cAMP-independent, stimulation of phosphatidylinositol 3-kinase activity in HL-60 cells. PMID- 9419818 TI - S49 cells endogenously express subtype 2 somatostatin receptors which couple to increase protein tyrosine phosphatase activity in membranes and down-regulate Raf 1 activity in situ. AB - S49 cells expressed type 2 somatostatin receptors (sstr2) by immunoblotting. Analysis by reverse transcription and polymerase chain reaction (RT-PCR) methodologies showed that S49 cells express predominantly sstr2A and sstr2B mRNAs; other subtypes were either not detected, in the case of sstr1, sstr3, sstr4, or variably detected, in the case of sstr5. No mutations were present in S49 cells at codon 12, 13, or 61 of the N-, K-, or H-ras genes. Nevertheless, randomly growing S49 cells contained Raf-1 activity by specific immune complex kinase assays. Treatment of S49 cells with somatostatin transiently inactivated the basal activity of Raf-1, but not that of B-Raf. Addition of somatostatin plus guanyl-5'-yl imidodiphosphate (GMPPNP) to S49 membranes stimulated PTPase activity. The concentration dependence for stimulation of PTPase activity correlated with high affinity binding of [125I-Tyr11]somatostatin-14. Both the effect of somatostatin to stimulate PTPase activity and to inactivate Raf-1 were abrogated by PTx. PTPase activity stimulated by somatostatin plus GMPPNP was recovered in a peak of high apparent M(r) (670,000) after solubilisation with Triton X-100 and Superose 6 chromatography. Furthermore, addition of activated, brain G alpha i/o subunits to fractions from control membranes stimulated PTPase activity in the high M(r) peak. Thus, S49 membranes contain a G-protein regulated PTPase (PTPase-G), and PTPase-G in these cells may reside in a high molecular weight complex. PMID- 9419819 TI - Sudden death in young people with apparently isolated mitral valve prolapse. AB - Ventricular electrical instability and sudden death in mitral valve prolapse (MVP) have been related to mitral valve regurgitation and left ventricular dysfunction, autonomic nervous system abnormalities, or underlying cardiomyopathy. The aim of the present study was to assess the frequency, nature and pathophysiologic significance of histologic myocardial abnormalities in young patients with apparently isolated MVP and sudden cardiac death. Among 163 cases of sudden cardiovascular death in young people, MVP was the only cardiac pathology found at postmortem gross examination in 17 (10%) (12 females and 5 males) aged 14 to 35 years, mean 24. In 12 cases sudden death occurred at rest (during pregnancy in 2). MVP was diagnosed during life in 8 patients, 6 of whom had experienced palpitations and/or syncope, and 3 had premature ventricular beats. In every case, postmortem gross examination revealed "floppy" mitral valve leaflets with marked myxoid degeneration, and no other cardiovascular pathology. Cardiomegaly with left chamber enlargement was observed in 5 cases (mild in 3 and moderate in 2). In 12 cases (70%), histopathologic study disclosed myocardial abnormalities which consisted of focal myocardial atrophy and fatty replacement of the right ventricular wall (mostly the outflow tract) in 9 cases, left ventricular myocardial disarray (without hypertrophy) in 2, and lymphocytic infiltrates in one. "Foetal" dispersion of specialized atrioventricular junction and fasciculoventricular Mahaim's fibers were found in 2 cases. In conclusion, apparently isolated MVP was found in nearly 10% of sudden cardiovascular fatalities in young people. Most young sudden death victims with MVP were asymptomatic females without significant mitral valve regurgitation. In more than two-thirds of the cases, histopathologic examination evidenced underlying silent but potentially arrhythmogenic myocardial substrates, mostly consisting of segmental right ventricular cardiomyopathic changes. PMID- 9419820 TI - Predictive value of heart rate in dilated cardiomyopathy treated with metoprolol. AB - Predictive factors of a favourable response to beta-blocker therapy are still unknown and the role of heart rate remains controversial. AIM: To investigate the relation between heart rate and the response to chronic metoprolol treatment in patients with dilated cardiomyopathy (DCM). METHODS: Ninety-eight consecutive patients with DCM, left ventricular ejection fraction (LVEF) < or = 0.40 and blood pressure < or = 140/90 mmHg were treated with metoprolol, associated with digitalis, diuretics and ACE-inhibitors. After 24 +/- 6 months, 48 patients (49%) were classified as "improved" on the basis of a clinical/instrumental score. RESULTS: Rest, mean 24-hour and maximal exercise heart rate were all significantly and directly related to the probability of improvement, but heart rate at rest, supine and in upright position, showed the highest predictive power. The relationship between heart rate and improvement with metoprolol appeared to be non-linear, with an increasing probability in patients with higher heart rate, but with a fall of the slope in cases with extreme tachycardia. By dividing our study population on the basis of the most important clinical variables, this complex relation was evident only in patients at a more advanced stage of the disease. CONCLUSION: Our analysis confirms the strict relationship between heart rate and improvement with chronic metoprolol therapy in patients with DCM. This relation seems to be non-linear and is influenced by the severity of the disease. PMID- 9419821 TI - Abnormal myocardial glucose handling in patients with syndrome X: effect of beta adrenergic blockade. AB - The present study was designed to test the hypothesis that patients with angina, positive exercise test and angiographically smooth coronary arteries (syndrome X), may exhibit abnormal myocardial glucose handling, as assessed by fluorodeoxyglucose (FDG) and positron emission tomography (PET) and to investigate the possibility that this abnormality could be reversed by treatment with betablockers, the drugs of choice in most patients with syndrome X. Eight consecutive patients (4 females, age 53 +/- 4 yrs) with syndrome X were studied. Off therapy, they underwent stress/rest 99m-TcMIBI SPET (360 degrees) and assessment of resting glucose metabolism by PET. PET studies were again performed after a 10 day treatment period on oral atenolol (100 mg/o.d.). All patients exhibited significant fasting FDG uptake in 2 or more myocardial regions. Overall, there were 28 of 48 segments (58%) with abnormal tracer uptake. On atenolol, there were only 5 segments with persistent FDG uptake (10%) in 2 patients. At rest, 7 patients exhibited perfusion defects in 14 myocardial segments. With stress performed in pharmacological wash-out, 5 patients developed perfusion defects in 10 myocardial segments. Eight of these segments were already underperfused at rest, and showed further reduction in perfusion after stress. All hypoperfused segments showed abnormal FDG uptake when the PET study was performed off therapy. The results suggest that, in patients with syndrome X, imbalance of the demand/supply ratio, either caused by limited coronary flow reserve or by primary vasoconstriction with reduction in myocardial perfusion, may determine a sustained metabolic shift towards anaerobic glycolysis. The mechanism by which atenolol improves metabolism in these patients could be simply related to reduction in O2 consumption. PMID- 9419822 TI - Italian experience with automated external defibrillators (AED). AB - In order to achieve widespread use of automated external defibrillators (AEDs) in Italy, we evaluated several models of AEDs in different clinical and artificial settings. We enrolled 268 consecutive patients with various rhythms and arrhythmias. Among these, 129 patients were referred to two different hospitals and 139 were enrolled by the pre-hospital care providers. AED was applied in 209 patients without symptoms of cardiac arrest and in 59 patients with cardiac arrest. The AEDs exhibited a 100% specificity (no false positives in 220 patients with non-shockable rhythm). Sensitivity was 92.3% (4 false negatives and 48 true positives in patients with VT/FV). This study confirms the absolute clinical safety and the high level of diagnostic reliability offered by the AEDs that were tested. PMID- 9419823 TI - Subintimal stent implantation for the treatment of a chronic coronary occlusion. AB - Subintimal passages of the guide wire during mechanical recanalization of chronic coronary occlusions are frequent and may result in the inability to reestablish anterograde flow in the distal coronary lumen. By using coronary stents, a conduct through the subintimal pathway can be obtained, allowing long-term restoration of the anterograde blood flow. We report on a case of a long, subintimal vessel reconstruction of a five-year-old coronary occlusion. Under intravascular ultrasound guidance, multiple coronary stents were implanted and good procedural and long-term clinical and angiographic results were achieved. PMID- 9419824 TI - Exacerbation of chronic obstructive lung disease mimicking acute myocardial infarction. PMID- 9419825 TI - Detection of myocardial viability with F-18-fluorodeoxyglucose and single photon emission computed tomography. AB - The application of 18-F-fluorodeoxyglucose (FDG) imaging with single photon emission computed tomography (SPECT) to detect tissue viability has been introduced recently in several centers, both in Europe and the USA. This paper summarizes the available data on FDG SPECT. Three studies have performed a direct comparison between FDG SPECT and FDG PET, showing good agreement between these techniques. Preliminary data in patients with poor left ventricular function undergoing revascularization suggest that FDG SPECT can predict improvement of ventricular function after revascularization. Comparative studies between FDG SPECT and low-dose dobutamine echocardiography also showed close agreement. Although the initial results are promising, larger studies are needed to determine the precise role of FDG SPECT in the assessment of myocardial viability. PMID- 9419826 TI - Current controversies in occupational medicine education in Europe. AB - The progressive implementation of the framework directive concerning occupational health and safety into che legislation of single European Union members is leading to changes in the role of occupational health professionals. It is important that Schools of Occupational Medicine contribute to training for competence and that occupational physicians continue to provide a high quality occupational health service in a changing environment. This could lead to a new and emerging role for the occupational physician, a key figure in prevention and health protection, capable not only to act as an expert but also as an advisor able to comply with employee and employer needs. The changing role of occupational physician will require new educational models which should allow the occupational physicians to be able to cope with problems and situations which can be encountered during their professional life. This will require a better harmonisation of educational and training programmes across the European Schools of Occupational Medicine. In fact, a variety of educational and training methods are currently adopted, although there is a substantial agreement about core values, core knowledge and core skills, which characterise the speciality. PMID- 9419827 TI - Partition coefficients of methyl tert-butyl ether (MTBE). AB - The knowledge of the partition coefficients between the air and arterial blood and between the blood and the different physiological compartments of the body are essential to the mathematical modelling of the respiratory uptake and elimination of toxic vapours. Partition coefficients of methyl tertbutyl ether (MTBE) in saline, olive oil, urine and human blood, and various rat tissues were calculated after gas-chromatographic quantification of MTBE in the air phase. The blood/air, urine/air saline/air, fat/air and oil/air partition coefficients (lambda) are respectively: 20.0, 15.6, 15.3, 142.0 and 138. PMID- 9419828 TI - Exposure to organic solvents and its effects on the central nervous system in workers of the Camacari petrochemical complex in Brazil. AB - While numerous studies have been carried out in industrialized countries, only very few epidemiologic investigations performed in developing countries are reported in the international literature. This study is one of the few examples of investigations carried out in a region where industrialization is at its beginning. A sample of 188 workers employed at the Camacari Petrochemical Complex in Bahia, Brazil, and exposed for over five years to organic solvents was submitted to neurobehavioral testing (QQS questionnaire, MANS battery) together with a 1/1 control group paired for age, school attendance and alcohol consumption. The exposed subjects showed differences significantly worse at emotional status, manual dexterity, recognition memory and subjective symptoms. Exposed subjects are therefore characterized by decreased psychophysical well being. PMID- 9419829 TI - Tolerability to prolonged lifting tasks. A validation of the recommended limits. AB - Prolonged physical exertion is subjectively regulated by the perception of effort. This preliminary study was conducted to validate the use of subjective perceptions of effort in assessing objectively tolerable workloads for prolonged lifting tasks. Ten healthy male subjects tested their maximal lifting capacity (MLC) on a lift dynamometer (LidoLift, Loredan Biomed., West Sacramento, CA) and underwent incremental and 30-minute endurance lifting tests. Cardiorespiratory parameters were monitored with an oxygen uptake analyzer, mechanical parameters were calculated using a computerized dynamometer. Ratings of perceived exertion were given on Borg's 10-point scale. Physiological responses to repetitive lifting were matched with subjective perceptions. A single-variable statistical regression for power functions was performed to obtain the individual "iso perception" curves as functions of the mechanical work exerted. We found that the "iso-perception" curve corresponding to a "moderate" perception of effort may represent the individual "tolerance threshold" for prolonged lifting tasks, since physiological responses at this level of intensity did not change significantly and the respiratory exchange ratio was less than one. The individually tolerable weight for lifting tasks lasting 30 min has been expressed as a percentage of the isoinertial MLC value and compared with the currently recommended limits for prolonged lifting tasks (Italian legislation D.L. 626/94). On the basis of our preliminary results a "tolerance threshold" of 20% MLC has been proposed for prolonged lifting tasks. PMID- 9419830 TI - Differential expression of estrogen receptor alpha and beta immunoreactivity by oxytocin neurons of rat paraventricular nucleus. AB - An understanding of the functional significance of the newly identified estrogen receptor (ER beta) in the brain will require definition of its expression pattern and relationship to ER alpha. Using an antibody generated against the C-terminus of rat ER beta, we report the presence of ER beta immunoreactivity in the lateral septum, medial amygdala, hippocampus and paraventricular nucleus (PVN) of ovariectomized rats. Double labelling studies in the PVN revealed that approximately 35% of oxytocin neurons located principally in the medial and lateral parvocellular divisions of the caudal PVN were immunoreactive for ER beta while vasopressin, somatostatin and magnocellular oxytocin neurons exhibited no ER beta staining with this antibody. No ER alpha immunoreactive cells were identified in the caudal PVN. These observations provide direct evidence for the differential expression of ER sub-types within neurons and indicate that ER beta may be of physiological significance in the regulation of hypothalamic parvocellular oxytocin neurons by estrogen. PMID- 9419831 TI - Implication of nitric oxide in NGF-induced cell differentiation: differences between neuronal and beta cells. AB - Both pancreatic beta cells (insulin-secreting cells) and neuronal cells express functional receptors for nerve growth factor. However, while the effect of nerve growth factor on neuronal differentiation is well known, its role on pancreatic beta cells is not established. It has been demonstrated that in PC12 cells, a well characterized NGF-responsive cell line, NGF increases the production of nitric oxide by inducing the expression of nitric oxide synthase. Nitric oxide is subsequently responsible for growth arrest, a step necessary for neuronal differentiation, visualized by the extension of neuronal-like processes. In the present study, we studied the effect of nerve growth factor on nitric oxide synthesis in INS-1 cells, an insulin-producing cell line which possesses the machinery necessary to respond to nerve growth factor. It was demonstrated that the expression of none of the three isoforms of nitric oxide was induced by nerve growth factor in INS-1 cells, strongly suggesting that nerve growth factor does not induce an increase in nitric oxide production in this cell line. Finally, we demonstrated that whereas growth arrest occurred in INS-1 cells cultured in the presence of a donor of nitric oxide (SNP), the simultaneous addition of SNP and nerve growth factor is not sufficient to induce the extension of neuronal-like processes in INS-1 cells. These dissimilarities strongly suggest that NGF plays a different role in neuronal and pancreatic beta cells. PMID- 9419832 TI - Endopeptidase EC 3.4.24.15 presence in the rat median eminence and hypophysial portal blood and its modulation of the luteinizing hormone surge. AB - The endopeptidase EC 3.4.24.15 (EP24.15) is a zinc metalloendopeptidase that is widely distributed in a variety of tissues, including the testes, pituitary and the central nervous system. Among its numerous roles in metabolizing and processing biologically-active peptides, the enzyme degrades gonadotropin releasing hormone (GnRH) by cleaving the central Tyr5-Gly6 bond. The aim of the present studies was to determine whether EP24.15 can modulate the concentrations of GnRH within the hypothalamo-hypophysial portal blood and thereby play a physiological role in reproduction. Our data suggest the presence of immunoreactive EP24.15 in the perivascular space of the median eminence and that this enzyme is secreted into portal blood. We have also shown a physiological role for this enzyme in that an inhibition of its activity with a specific inhibitor augmented the steroid-induced LH increase in ovariectomized rats. The present results suggest that secretory and post-secretory mechanisms are important in shaping the GnRH signal from the central nervous system; GnRH metabolism by EP24.15 may be one such mechanism. PMID- 9419833 TI - Immunohistochemical localization of novel CART peptides in rat hypothalamus, pituitary and adrenal gland. AB - CART peptide specific polyclonal antisera were raised in rabbits. The antisera were raised to CART peptide fragments that span most of the predicted CART protein. The specificity of each antisera was demonstrated by blockade of immunostaining by the immunizing peptide but not by the other CART peptide fragments. In the hypothalamus and pituitary of colchicine and noncolchicine treated rats, immunostaining was observed in cell bodies, fibers and varicosities. Clusters of cells were also stained in the adrenal medulla. It is noteworthy that cellular immunostaining was only found in areas previously shown to express CART mRNA. These findings indicate the presence of CART peptide(s) in the hypothalamus, pituitary, and adrenal gland. Furthermore, we also present evidence for the possible processing of the CART pro-peptide into smaller peptide fragments. These neuroanatomical findings suggest a role of CART peptides in hypothalamic, pituitary and adrenal function. PMID- 9419834 TI - Neuronal synchronization and ionic mechanisms for propagation of excitation in the functional network of immortalized GT1-7 neurons: optical imaging with a voltage-sensitive dye. AB - Immortalized gonadotropin releasing hormone (GnRH) neurons (GT1 cell line) in culture release GnRH in a pulsatile manner, suggesting that GT1 cells form a functional neuronal network. Optical imaging techniques and a voltage-sensitive fluorescent dye (RH795) were used to study the mechanism of neuronal synchronization and intercellular communication in cultured GT1-7 cells (one of the subclones of the GT1 cell line). The majority (79%) of GT1-7 cells in contact with one another revealed synchronized fluctuations in spontaneous neuronal activity. When a cell in contact with other cells was electrically stimulated, the evoked excitation was propagated to neighbouring cells. The ionic mechanisms involved in the propagation of electrical signals between interconnected GT1-7 cells were investigated using various blockers of Na+, Ca2+ and K+ channels. The propagation of stimulus-evoked excitation was prevented by the voltage-dependent Na+ channel blocker tetrodotoxin. It was also prevented by the voltage-dependent Ca2+ channel blockers, Ni+ (nonselective), nimodipine (L-type) and flunarizine (T type > L-type), but not apparently affected by omega-agatoxin IVA (P- and Q-type) and omega-conotoxin MVIIA (N-type). The propagation was not influenced by the K+ channel blockers, quinine, tetraethylammonium and Ba2+, but in some cases, it was enhanced by 4-aminopyridine (4-AP) and prevented by apamin. These results suggest that voltage-dependent Na+ channels and L- and T-type Ca2+ channels are involved in the propagation of electrical signals in the GT1-7 neuronal network. Ionic mechanisms, through 4-AP- or apamin-sensitive K+ channels, also seem to be involved in the regulation of signal propagation. These mechanisms may underlie the functioning of the neuronal network formed by immortalized GnRH neurons. PMID- 9419835 TI - Somatostatin plays a dual, stimulatory/inhibitory role in the control of growth hormone secretion by two somatotrope subpopulations from porcine pituitary. AB - Previous results from our laboratory demonstrated the existence of two subpopulations of porcine somatotropes of low- (LD) and high density (HD) that exhibit differences in ultrastructure and respond in an opposite manner to somatostatin (SRIF) in vitro. In LD cells, SRIF did not affect basal growth hormone (GH) release but partially blocked the stimulatory effect induced by GH releasing factor (GRF). Conversely, SRIF paradoxically stimulated the secretory activity of HD somatotropes. Here, we have analysed in detail the basic parameters that characterize this differential response. To this end, the time- and dose-dependent effects of SRIF-14 were evaluated on separate monolayer cultures of both subpopulations. Likewise, the direct effect of the peptide on individual somatotropes from each subset was assessed by cell immunoblot assay. Finally, we compared the effects of SRIF-14 and SRIF-28 on cultures of LD and HD cells. SRIF-14 (10(-7) M) induced a rapid (30 min) and sustained (4 h) 2-fold increase in GH release from HD cells, whereas it did not affect GH secretion from LD somatotropes. Surprisingly, a low dose of SRIF (10(-15) M) stimulated GH release from both LD (154.1 +/- 8.2% of basal, P < 0.05) and HD (337.2 +/- 55.5% of basal, P < 0.05) subpopulations, even more effectively than higher doses of the peptide. Results from cell blotting showed that SRIF stimulatory effects were exerted directly upon individual somatotropes. Finally, SRIF-28 elicited similar responses to those observed for SRIF-14 in both somatotrope subpopulations, yet 10(-15) M SRIF-28 was less potent than the same dose of SRIF-14 in stimulating GH release from HD cells. Our present findings demonstrate that SRIF can function as a true GH-releasing factor in cultures of porcine pituitary cells by acting specifically and directly upon somatotropes. Furthermore, together with previous observations, these results strongly suggest that SRIF is not merely an inhibitor of GH release in pigs, but might play a dual modulatory role. Heterogeneity of the somatotrope population contributes greatly to this divergent effect of SRIF. PMID- 9419836 TI - Melatonin inhibits naloxone-induced luteinizing hormone release in ovariectomized estrogen-primed rats. AB - The present study aimed to examine the effect of melatonin on naloxone-induced luteinizing hormone (LH) secretion in ovariectomized estrogen-primed rats. A single intracerebroventricular (i.c.v.) injection of naloxone (mu opioid receptor blocker, 15 micrograms) or an intravenous (i.v.) injection of LH-releasing hormone (LHRH, 50 ng/kg) elicited a transient and significant increase in the serum LH concentration within 10 min. While an i.c.v. injection of 100 ng melatonin by itself did not change the basal LH release, it almost completely inhibited the naloxone-induced LH release. Melatonin (10 ng) also significantly reduced the effect of naloxone. However, an i.c.v. injection of 100 ng melatonin did not affect the LHRH-induced LH release. In separate experiments, the effect of melatonin on naloxone-induced pulsatile LH secretion was studied in estrogen treated rats. A continuous i.v. infusion of naloxone (20 mg/kg/h) induced LH pulses in rats treated i.c.v. with saline. An i.c.v. administration of 100 ng melatonin, which by itself did not affect basal LH secretion, significantly reduced the frequency, but not the amplitude, of LH pulses induced by the naloxone infusion. These results show that melatonin has a suprapituitary site of action to inhibit naloxone-induced LH release, and suggest that melatonin has an effect in inhibiting the activity of the hypothalamic LHRH pulse generator, either directly or indirectly, in female rats. PMID- 9419837 TI - Changes in oxytocin receptor mRNA in rat brain during pregnancy and the effects of estrogen and interleukin-6. AB - Changes in brain oxytocin receptor (OTR) binding sites during the course of pregnancy may influence the sudden onset of maternal behavior in female rats at parturition. In situ hybridization was used to identify changes in OTR messenger ribonucleic acid (mRNA) expression during pregnancy and parturition. Two patterns of mRNA regulation were observed. Relative to diestrus virgin control females, OTR mRNA was elevated in the lateral septum and medial preoptic area at days 13 15 of pregnancy but not on the morning of parturition. In the central nucleus of the amygdala and ventromedial nucleus of the hypothalamus (VMH), OTR mRNA was most abundant on the morning of parturition. Strong signals for OTR mRNA were detected in the bed nucleus of the stria terminalis, hypothalamic paraventricular nucleus, supraoptic nucleus and suprachiasmatic nucleus. However no group differences were detected in these areas. As estrogen and interleukin-6 have been suggested to modulate OTR gene expression and both are elevated at the time of parturition, their effects on OTR mRNA in the brain were examined. Estrogen and interleukin-6, given simultaneously, significantly elevated the concentration of OTR mRNA in the VMH, but not in the amygdala. The increase in the VMH was significantly greater than that produced by estrogen alone, and interleukin-6 alone had no effects. These results demonstrate that transcriptional regulation of OTR gene expression mediates changes in receptor density in the brain in a region specific manner during pregnancy and suggests a potential mechanism for some of these changes. PMID- 9419839 TI - Age-dependent accumulation of hybrid vasopressin-oxytocin gene products but not hybrid oxytocin-vasopressin products in the endoplasmic reticulum of Brattleboro rats. AB - The age-dependence of the incidence of magnocellular neurosecretory neurons containing abnormal accumulations of peptide in the rough endoplasmic reticulum was examined in homozygous Brattleboro rats and in their wild-type Long Evans counterparts. Neurons in which the immunophenotype of the peptide aggregates indicate that somatic cross-over mutations involving the 5' end of the vasopressin gene and the 3' end of the oxytocin gene have occurred, increased with age in homozygous Brattleboro rats, reaching a maximum of 24 cells per hypothalamus (approximately 0.6% of the vasopressin neurons). The increase occurred in both male and female animals but was significantly greater in females. The average incidence of such cells was 6 times greater in the supraoptic than in the paraventricular nucleus. No such cells could be detected in either nucleus of Long Evans rats despite the evidence for hybrid mRNA in these animals. Moreover, no accumulation of peptide translated from the hybrid mRNAs derived from the 5' end of the oxytocin gene and the 3' end of the vasopressin gene could be detected in either Brattleboro or Long Evans animals. These results strongly suggest that the accumulation of peptide in the rough endoplasmic reticulum of vasopressin neurons in homozygous Brattleboro rats is due to an abnormality other than the somatic crossing-over mutation. A second type of abnormal magnocellular neuron with accumulations of peptide in the rough endoplasmic reticulum, in which the immunophenotype of the peptide reveals products derived only from the oxytocin precursor, was present in both Long Evans and Brattleboro rats, but did not increase with age in Brattleboro rats. The incidence of these cells was similar in the supraoptic and paraventricular nuclei. PMID- 9419838 TI - Hypoglycaemia-induced inhibition of pulsatile luteinizing hormone secretion in female rats: role of oestradiol, endogenous opioids and the adrenal medulla. AB - Oestradiol (E2) has been shown to exacerbate the inhibitory effect of hypoglycaemic stress on gonadotrophin-releasing hormone pulse generator (GnRH) activity in primates. The mechanism by which this is mediated is not yet known. We therefore aimed to establish whether there is a sensitizing influence of E2 on the suppression of LH pulsatility in response to hypoglycaemia in the female rat, thus providing a more amenable model in which to study this phenomenon. In ovariectomized Wistar rats with E2 replacement, insulin-induced hypoglycaemia (0.5 U/kg i.v.) resulted in an interruption of pulsatile LH secretion. Induction of the same degree of hypoglycaemia in ovariectomized rats without E2 replacement was without effect on LH pulsatility. Naloxone administration prevented the hypoglycaemia-induced inhibition of LH pulses. Because hypoglycaemia is a potent activator of the sympathetic nervous system, we also tested the hypothesis that the adrenal medulla is involved in this suppression of LH pulses in the rat. Adrenomedullectomy completely prevented this inhibitory response to hypoglycaemic stress. These data are consistent with the hypothesis that E2 sensitizes the GnRH pulse generator to the inhibitory influences of hypoglycaemic stress in the rat. Furthermore, a clear role for both endogenous opioid peptides and the adrenal medulla in the stress-induced suppression of LH pulsatility is identified. PMID- 9419840 TI - Antimalaric effect of an alcoholic extract of Artemisia ludoviciana mexicana in a rodent malaria model. AB - Chloroquine resistance of Plasmodium falciparum first and of P. vivax more recently, stimulated the search for new antimalarics. Chinese investigators have introduced new compounds obtained from extracts of Artemisia annua which possess an antimalaric active principle different from those of the drugs in use. In Mexico eight species of Artemisia have been described and among them just A. ludoviciana has been empirically used in the treatment of intermittent fever. To know whether mexican Artemisia had antimalaric activity several in vivo experiments were performed. Different type of extracts from two Artemisia species were prepared and assayed in five different doses on mice infected by Plasmodium yoelii yoelii, in a four-day test scheme. Here, only the results of the assays on ethanolic extract of A. ludoviciana are presented. The results of the in vivo experiments showed that the parasite reproduction was inhibited up to 98.6% at the fifth day, as compared with the controls; the ED50 was of 29.2 mg/kg and the SM50 of 28.7. We looked after the presence of artemisinin in the ethanolic extract, without success. PMID- 9419841 TI - Phagicola longa (Heterophyidae) in dogs from Chile: morphological findings and taxonomical problems. AB - Trematodes found in the intestines of dogs from Chillan, VIII region of Chile, were studied. The species identified was Phagicola longa, a fluke of the Heterophyidae family, parasites of fish-eating birds or mammals in several countries. P. longa is very similar to P. arnaldoi, a species described in dogs from South America. The number of vitelline follicles and the distribution of cuticular spines are the only distinctive features for the two species. However, many authors report the same number of vitelline follicles for P. arnaldoi and P. longa. Furthermore, the distribution of cuticular spines is very similar. P. longa has a small area devoid of spines when compared to P. arnaldoi but a wide variability of this generic character in the Heterophyidae was observed. As previously reported, P. arnaldoi could be a recent synonym of P. longa, which is the only species of Phagicola that can be found in South American dogs. This is in agreement with the study of metacercariae found in Mugilidae, the intermediate hosts of Phagicola, from American waters in which P. longa is the only species found. The authors emphasize the finding of this trematode in dogs, since it could possibly cause also human infection. PMID- 9419843 TI - New record of Nosopsyllus (Nosopsyllus) barbarus (Jordan and Rothschild, 1912) (Aphaniptera: Dolichopsyllidae) from Sicily. Morphology at scanning electron microscopy of N. (N.) barbarus and N. (N.) fasciatus (Bosc D'Antic, 1800). AB - Nosopsyllus (N.) fasciatus, species or complex cosmopolitan and very polymorphic, is established in most of Italian regions, in association with murine rodents, especially domestic species. N. (N.) barbarus, similar to N. fasciatus in modified segments of the male and perhaps in ecology, is presently known from North Africa regions and a few Mediterranean islands; in Italy it is reported from Sardinia and Egadi islands. In this note the N. barbarus presence in Sicily (Palermo) and the contemporary record of N. fasciatus from the same country and host species (Rattus rattus frugivorus) are reported for the first time. The coexistence of both species was shown until now, only in Sardinia. A scanning electron microscope was used to describe the clasper of the male of N. fasciatus and the main diagnostic surface differences between females of N. fasciatus and N. barbarus. PMID- 9419842 TI - Ivermectin and moxidectin in two filarial systems: resistance of Monanema martini; inhibition of Litomosoides sigmodontis insemination. AB - Effects of ivermectin and moxidectin were compared on two filarial species: Monanema martini which presents dermal microfilariae and induces Onchocerca-like lesions in its natural murid host Lemniscomys striatus, and Litomosoides sigmodontis (= L. carinii). M. martini microfilariae showed an unusual resistance to ivermectin, in vitro and in vivo; moxidectin was no more efficient. However, the two drugs used at high concentrations deeply altered the uterine embryogenesis, but had no lethal effect on adult filariae. L. sigmodontis blood microfilariae showed a great susceptibility to moxidectin, similar to that previously described for ivermectin. The two drugs also induced a long term effect because they inhibited the insemination of the female filariae. This result reinforces the observations made by other authors on the human parasite, Onchocerca volvulus. PMID- 9419844 TI - Atypical P. vivax and P. ovale morphology in two imported cases of malaria in Spain. AB - The finding of two imported cases of malaria in Spain is reported. Although their origin, their age and their clinical history were different, they both showed parasites of atypical morphology. One was diagnosed as P. vivax and the other as P. ovale, but the morphology observed in both cases does not correspond to what is considered as normal in the two species. In the first case forms similar to those of P. ovale were observed and in the second case P. vivax-like forms. The findings reported confirm the overlapping morphological variability of the two species. PMID- 9419845 TI - Acanthamoeba keratitis associated with contact lenses; report of three cases in Italy. AB - Three female patients, aged from 17 to 44 years, developed Acanthamoeba keratitis associated with the use of soft contact lenses and cosmetic contact lens. Two of these patients were myopic and wore soft contact lenses for optical purpose, while the third patient used a cosmetic lens. In this paper we describe the related risk factors, the success of cultural procedures and the outcome of medical management. In 2 out of the 3 cases we obtained cultures positive for Acanthamoeba, from the contact lenses and the contact-lens case. The isolated strains belong to the group II according to Pussard and Pons and they were proven pathogen for experimentally infected mice. All infections were treated successfully; however in 2 out of the 3 patients the visual acuity was reduced to the light perception. PMID- 9419846 TI - Spatial distribution and seasonality of ticks (Acarina: Ixodidae) in a protected area in the northern Apennines. AB - Infestation of small rodents by ixodid ticks and frequency of host-seeking ticks collected by dragging were studied at Orecchiella Natural Park (Northern Apennines) in 1994 and 1995. Levels of infestation of Apodemus spp. by immature Dermacentor marginatus were higher in beech wood (5.1 larvae and 1.3 nymphs per mouse) than in oak-chestnut wood and in coniferous wood. Larval D. marginatus peaked in mid summer, whereas nymphs peaked in late summer. Host-seeking Haemaphysalis punctata were mostly found on south-facing limestone rocks with scarce vegetation (7.8 larvae/km dragging). Conversely, D. marginatus larvae were most frequent in wooded areas (3.2 larvae/km dragging). Ixodes ricinus was rare in the Park, and Borrelia burgdorferi was not isolated from ear punches collected from 122 small rodents. PMID- 9419847 TI - Epidemiology of hydatid disease in the Mediterranean basin with special reference to Italy. AB - Hydatidosis, caused by dog tapeworms of the genus Echinococcus, is one of the most important cestode infections in man. Of the four species of the genus Echinococcus, only Echinococcus granulosus is widespread in the Mediterranean basin which is still, unfortunately, a major endemic focus. This article critically reviews its diffusion in humans and sheep over the past twenty years. Although most of the available data are scarce, fragmentary, and not homogeneous, they represent the only possibility at present of evaluating the parasitic pressure. PMID- 9419848 TI - Factors affecting trypanosome infection rate in tsetse fly (Diptera: Glossinidae) populations. AB - Wide variations in trypanosome infection rate are observed in different tsetse fly (Glossina spp.) populations Environmental factors and features proper to the vector, mammalian host and infecting-trypanosome species acting in the acquisition and development of infective-trypanosome infection in tsetse are examined. PMID- 9419849 TI - Ixodes ricinus (Acari: Ixodidae) infestation on roe deer (Capreolus capreolus) in Trentino, Italian Alps. AB - The most important tick-deer system potentially supporting the epidemiology of Lyme disease in the Italian Alps is that regarding Ixodes ricinus (L.) and roe deer (Capreolus capreolus L.). In this study, the pattern of tick infestation on 562 male roe deer harvested in September 1994 in 56 game districts of Trentino, Northern Italy, was assessed. The prevalence and density of infestation by I. ricinus were analyzed by a model based on classification and regression trees (CART), using both discrete and continuous variables concerning environmental and host parameters. The model discriminated attitude and host density as the 2 variables having the greatest effect on the prevalence and density of infestation of deer; the levels of infestation were higher at an altitude below 1125 m or at roe deer densities over 8.5 head per 100 ha. The density of tick infestation tended to be higher in older roe deer. PMID- 9419850 TI - Transmission of mixed malaria species and strains by mosquitoes, as detected by PCR, in a study area in Guinea-Bissau. AB - Parasites present in blood samples of asymptomatic carriers and in the midgut of mosquitoes collected within a few days from the same households, have been analysed by PCR. A high prevalence (32%) of infected mosquitoes was observed and, in half of these, two parasite species were found simultaneously. The distribution of parasite species in the mosquito correlated with that found in the infected persons. Genotype patterns of Plasmodium falciparum populations were however found to be different in the two sets of samples. These results and the potential of PCR are discussed with reference to investigations of the dynamics of malaria transmission. PMID- 9419851 TI - Phylogenetically distant intracellular symbionts in termites. AB - Cockroaches are known to harbour intracellular bacteria in specialised cells (mycetocytes, or bacteriocytes) of the fat body. In termites, mycetocyte bacteria have been observed only in Mastotermes darwiniensis. These symbionts are thought to have originated from a bacterium that infected an ancestor common to cockroaches and termites. Thus, loss of the infection should have occurred during evolution in all termite lineages, with the exception of that leading to M. darwiniensis. One might suspect that traces of the ancient infection may be present in some termites, in the form of non-mycetocyte intracellular bacteria (e.g. a small number of bacteria within normal cells). Indeed, circumstantial evidence for the presence of intracellular bacteria in two termite species has been reported. However, no data are available on the actual distribution of these bacteria in termites, or on their relationships with the mycetocyte bacteria of cockroaches and M. darwiniensis. In this paper we report results indicating that non-mycetocyte intracellular bacteria are widespread in termites. These results were obtained by electron microscopy on representatives of nine termite species. In addition, sequence analysis of the 16S rRNA genes indicated that the non mycetocyte bacteria of termites belong to the wolbachia group of the alpha-2 subclass of the proteobacteria. These latter bacteria are not related to the mycetocyte bacteria of cockroaches and M. darwiniensis, which belong to the blattabacterium group of the flavobacteria-bacteroides. PCR analyses with primers specific for wolbachia or blattabacterium provided further support for the identification of the observed non-mycetocyte bacteria as members of the wolbachia group. PMID- 9419852 TI - Rapid microbiological methods with hazard analysis critical control point. AB - The proactive approach to ensuring food safety termed hazard analysis critical control point (HACCP) was introduced in the 1960s by the Pillsbury Company, in collaboration with the U.S. Army Natick Laboratories and National Aeronautics and Space Administration, to help guarantee that astronauts would not be incapacitated by the trauma of foodborne illness during space flights. The approach has subsequently been adopted as the standard food safety management system world-wide and is seen as forming the basis for harmonization of food inspection regulations necessitated by trade agreements such as General Agreement on Tariffs and Trade and North American Free Trade Agreement as the move toward globalization of trade in food products gains momentum. The new U.S. Department of Agriculture Mega-Reg requires mandatory introduction of HACCP, and the Food Safety Enhancement Program of Agriculture and Agri-food Canada, as well as the "due diligence" legislation of the European Union, is centered on HACCP principles. PMID- 9419853 TI - Liquid chromatographic determination of tilmicosin in swine feed at 200-400 mg/kg level: interlaboratory study. AB - An analytical method for the determination of tilmicosin at 200-400 mg/kg, the intended use concentration range, was evaluated in an interlaboratory study involving 5 laboratories, including the sponsor. The interlaboratory study evaluated the intra- and interlaboratory precision and accuracy of a tilmicosin feed method. The method procedure involved extracting tilmicosin from feed by adding 200 mL extractant to 20 g feed and shaking for 1 h. The extract is filtered and analyzed by gradient liquid chromatography which separates tilmicosin from feed matrix in 30 min. Each laboratory assayed 5 replicates of fortified feed at concentrations of 0, 100, 200, 400, and 600 mg/kg. The mean recovery among fortified samples ranged from 81.4 to 98.8%, with a percent coefficient of variation (%CV) ranging from 0.3 to 4.0%. For all blank control feed samples no significant interferences were observed. In addition, each laboratory assayed 5 replicates of medicated feed samples prepared at 2 levels (200 and 400 mg/kg) with either a horizontal or vertical mixer. Along with the medicated feed samples were included 5 replicates of a blank control feed. The identities of the medicated and blank control feed samples were blinded to the analysts. The results for the medicated feed samples ranged from 95.8 to 106% of label claim, with a %CV ranging from 2.1 to 6.7%. PMID- 9419854 TI - Determination of tilmicosin in swine feeds by liquid chromatography. AB - This method determines tilmicosin in feeds over a concentration range of 100 to 600 mg/kg. Tilmicosin is extracted from swine feeds by adding 200 mL of a swine feed extractant (20 + 80, acetonitrile-Millipore water, pH 2.5, with 25 mM dibutylammonium phosphate) to 20 g feed and placing on a shaker table for 1 h. This extractant is filtered and analyzed by liquid chromatography (LC). A gradient LC method is used to separate tilmicosin from the feed matrix in 30 min of run time. The recovery of tilmicosin from fortified feeds ranged from 96.7 to 112%, with the coefficients of variation (CVs) ranging from 1.4 to 3.9%. The determination of tilmicosin in medicated feeds resulted in an average recovery of 92.7% of labeled claim for pelleted feeds at 200 mg/kg and 99.1% of labeled claim for mash feeds at 400 mg/kg. Determination of tilmicosin in medicated feeds resulted in CVs ranging from 2.6 to 3.8%. The method has shown no interference with 18 other drugs. PMID- 9419855 TI - Planar chromatography for quantitative determination of ampicillin residues in milk and muscle. AB - A method involving protein precipitation with trichloroacetic acid and ampicillin hydrolysis with sodium hydroxide was developed for determination of ampicillin residue in milk and muscle. The hydrolysis product reacts in an acidic medium, in the presence of mercuric chloride, to form a fluorescent compound, which is extracted from the matrix with dichloromethane and concentrated on a silica Sep Pak cartridge in a single step. After elution of the silica cartridge and evaporation of the solvent, the residue is recovered in 100 microL ethyl acetate, and 20 microL is deposited on a chromatoplate and chromatographed with ethyl acetate. The method allows determination of ampicillin residues in milk and muscle fortified at 4 micrograms/L and 50 micrograms/kg, respectively. PMID- 9419856 TI - Determination of nicarbazin in eggs by liquid chromatography-atmospheric pressure chemical ionization mass spectrometry. AB - A method was developed to determine in eggs 2 components [4,6-dimethyl-2 hydroxypyrimidine and 1,3-bis(4-nitrophenyl)urea] of the anticoccidial drug nicarbazin, used to treat poultry. Samples were extracted with acetonitrile, and the extracts were washed with hexane and evaporated to dryness before analysis by liquid chromatography/mass spectrometry with atmospheric pressure chemical ionization. By switching from positive to negative ion monitoring and using gradient elution, both components were measured within one run. The limit of quantitation of the assay was 10 ng/g for each component. The results of a preliminary feeding trial in which chickens were fed contamination levels of the drug are also reported. PMID- 9419857 TI - Determination of tilmicosin in bovine and porcine sera by liquid chromatography. AB - A liquid chromatographic (LC) assay is described for determining tilmicosin in bovine and porcine blood sera. Tilmicosin is isolated from the serum matrix and purified by solid-phase extraction with C18 sorbent. Sample is analyzed by LC using a gradient system with a phenyl reversed-phase column that separates tilmicosin from the matrix in 30 min. Tilmicosin is measured by UV absorbance at 280 nm. Validation of assay included evaluation of accuracy, precision, linearity, specificity, sensitivity, range, and sample stability. The method has a limit of quantitation of 0.1 ppm and a validated range of 0.1 to 10.0 ppm. Recoveries were 91-95% for bovine serum and 85-93% porcine serum. The limit of detection was 0.05 microgram/mL. Limits of detection and quantitation were based on 3 and 6 times the baseline noise of control serum samples, respectively. Relative standard deviations of precision samples (n = 6) were 2% or less for both sera. The method has better specificity and analysis time than previous microbiological methods for tilmicosin in sera. PMID- 9419858 TI - Survival and detection of Shigella flexneri in vegetables and commercially prepared salads. AB - The normal bacterial microflora of 4 commercially prepared salads (coleslaw, crab salad, carrot salad, and potato salad) and 3 vegetables (green pepper, onion, and cabbage) were evaluated. Twenty-eight species of bacteria, including potential pathogens, were isolated. The foods were artificially inoculated with an avirulent mutant strain of Shigella flexneri 5 (pHS1059) to develop a method for the rapid detection of Shigella spp. Bacteria were separated from insoluble and particulate salad ingredients by filtration through shark skin filter paper and by low speed centrifugation. S. flexneri survived at 4 degrees C in all salads for at least 11 days and up to 20 days in crab salad. The polymerase chain reaction (PCR), using primers for amplification of a 118-base pair DNA fragment from the virulence-associated spa region, present in all Shigella spp., was used to detect S. flexneri in filtrates from salads inoculated with S. flexneri 5 (pHS1059). DNA was amplified from all of the artificially contaminated salads and vegetables except green pepper. After 3-5 days of storage, the PCR also amplified S. flexneri DNA from salads that had been enriched with nutrients to increase the number of bacteria. Green peppers contained a PCR inhibitory substance that was attenuated by dilution and enrichment before the PCR. No amplification of DNA was observed in foods to which S. flexneri had not been added. PMID- 9419859 TI - Effects of salt and serum on the sporicidal activity of liquid disinfectants. AB - This study compares the effects of various concentrations of salt or serum in the killing of Bacillus subtilis spores by either glutaraldehyde, sodium hypochlorite, cupric ascorbate, hydrogen peroxide, peracetic acid, formaldehyde, or phenol. Salt affected only glutaraldehyde, its sporicidal activity increasing with an increase in concentration of sodium bicarbonate or sodium chloride. The sporicidal activity of glutaraldehyde was minimal when the concentrations of aldehyde groups and lysine residues from protein were similar. We present an equation describing the effect of serum on spore survival as a function of glutaraldehyde concentration that fits the data with a regression coefficient of 0.9. Cupric ascorbate and peracetic acid were inhibited by serum, but this effect was linked to a rise in pH. Sodium hypochlorite was the agent most sensitive to protein, with its sporicidal activity nearly disappearing in the presence of 2% serum or an equivalent amount of purified protein. PMID- 9419860 TI - Comparison of antibody-direct epifluorescent filter technique with the most probable number procedure for rapid enumeration of Listeria in fresh vegetables. AB - The antibody-direct epifluorescent filter (Ab-DEFT) technique was evaluated as a rapid alternative to the most probable number (MPN) method for enumeration of artificially inoculated Listeria monocytogenes in ready-to-eat packaged salads and other fresh vegetables. Ab-DEFT was performed by homogenization of food in mesh-lined Stomacher bags, followed by prefiltration of homogenate through a 5 microns pore nylon filter, and passage of filtrate through a 0.4 micron pore black polycarbonate filter to collect and concentrate Listeria cells. After cells were stained with a fluorochrome-labeled polyclonal antibody to Listeria, the filter surface was examined by epifluorescence microscopy, and fluorescent cells were counted. A 3-tube MPN procedure was performed by successive enrichments of homogenized foods in Listeria enrichment and Fraser broths, followed by selective plating. Ab-DEFT provided quantitative determinations of Listeria cells that correlated with plate counts and MPN estimates in a linear response over a range of cell concentrations from 10 to 10(7) colony forming units (CFU)/mL. Microbial backgrounds as high as 10(8) CFU/mL did not affect performance of Ab-DEFT. In contrast to the MPN method, which required 5 days to perform, quantitation by Ab DEFT could be completed in less than 1 h. Despite cross-reactivities demonstrated by the polyclonal fluorescent antibody, the potential of Ab-DEFT as a rapid alternative to MPN for microbial cell enumeration was evident. PMID- 9419861 TI - Analysis of aflatoxins in poultry and pig feeds and feedstuffs used in Colombia. AB - Feedstuffs and mixed feeds used for poultry and pig nutrition in Colombia were analyzed for aflatoxins by using a liquid chromatographic technique with a limit of detection of 1 microgram/kg for each aflatoxin (B1, B2, G1, and G2). Samples of grain sorghum, maize, processed soybean, rice meal, cottonseed meal, and poultry and pig feeds, representative of Colombian production for the 1995-1996 harvest, were taken from feed-manufacturing plants in various cities. Aflatoxins were detected in 11 of 45 samples of sorghum, 4 of 33 samples of maize, 8 of 22 samples of rice meal, 15 of 17 samples of cottonseed meal, 1 of 12 samples of other feedstuffs, 12 of 30 samples of poultry feed, and 7 of 16 samples of pig feed. Aflatoxins were not detected in soybean. Only 9 of 58 positive samples contained total aflatoxin levels exceeding maximum tolerable limits in Colombia. PMID- 9419862 TI - Induction and characterization of multianalyte antibodies against penicillins in egg yolk. AB - Antibodies against penicillins were induced in eggs of laying hens after immunization with 6-aminopenicillanic acid (6-APA) coupled to key-hole limpet hemocyanin (KLH). Development of the antibody titer was monitored by an indirect enzyme-linked immunosorbent assay (ELISA), with 6-APA coupled to ovalbumin as antigen for coating microtiter plates. Different characteristics (time course, affinity) of the immune reaction were observed by testing eggs of individual hens. Titer values varied between 150 and 2000. Antibodies were isolated by polyethylene glycol precipitation and affinity chromatography, using a hapten sorbent with 6-APA as ligand. Glycine buffer, pH 3.0, was used to elute the immunoglobulins. Antibody specificity was determined in a competitive ELISA with 7 penicillins and the cephalosporin cephalexin as competitors. Cross reactivities for the penicillins were between 100 and 290% (6-APA = 100%). Cephalexin cross reacted only marginally (3%). PMID- 9419863 TI - Determination of aflatoxins in beer. AB - Aflatoxins B1, B2, G1, and G2 were determined at parts-per-trillion levels in beer by immunoaffinity column cleanup and reversed-phase liquid chromatography (LC) with fluorescence detection after trifluoroacetic acid derivatization. Silanized vials were necessary for the evaporation step in order to obtain good recoveries of aflatoxins from spiked beer samples. Recoveries averaged 90-104%, 94%, 84-87%, and 89% for aflatoxins B1, B2, G1, and G2, respectively, at levels of 9.7-133 ng B1, 46 ng B2, 35-140 ng G1, and 41 ng G2/L. Detection limits were 19-20 ng/L for aflatoxins B1 and G1 and 15-16 ng/L for aflatoxins B2 and G2 (signal-to-noise ratio = 3:1) obtained by using an excitation wavelength of 360 nm; at 340 nm these detection limits were lowered to about 2 ng/L. Analysis of 24 beer samples, the majority from the United States and Mexico, showed natural contamination of one sample of Mexican beer at 49 ng B1/L when determined at 360 nm excitation, but reanalysis of 23 of the samples using 340 nm excitation indicated that an additional 4 Mexican samples and one Brazilian sample contained aflatoxin B1 at low levels (< 10 ng/L). PMID- 9419864 TI - Determination of fish flesh content in frozen coated fish products (modification of AOAC Official Method 971.13): Collaborative Study. AB - An intralaboratory collaborative study evaluated a modified version of AOAC Official Method 971.13 for determining the fish flesh content (FFC) in frozen coated fish products by comparing it with the on-line method. Eleven collaborators analyzed 36 products (a total of 6336 test samples). Each product targeted one of 4 percent fish flesh (PFF) levels (35, 50, 65, and 80). Products were manufactured from one of 3 raw materials (fillet blocks, minced blocks, and natural fillets) and processed in one of 4 forms (sticks, portions, formed portions, and fillets) and one of 4 styles (raw breaded, batter-dipped, precooked, and fully cooked). Each "official" test sample was tracked through the processing system and weighed (1) before battering and/or breading and, depending on product style, before frying; and (2) after battering and/or breading and, depending on product style, after frying; so that it served as its own control. These weights were used to calculate actual percent fish flesh (APFF) and considered to be generated by the on-line method. Collaborators weighed official test samples (1) before scraping; and (2) after scraping. These weights were used to calculate determined percent fish flesh (DPFF) and considered to be generated by the modified AOAC method. APFF and DPFF were the primary data for statistical analysis. Recoveries ranged from 71.75 to 106.40%. Repeatability (method precision indicator for a single collaborators) relative standard deviation (RSDr) values ranged from 1.04 to 8.37%. Corresponding reproducibility (method precision indicator among collaborators) relative standard deviation (RSDR) values ranged from 1.41 to 11.95%. The DPFF mean was lower than the APFF mean for 30 (83.3%) of 36 products. For 29 of these 30 products, the differences between method means (APFF minus DPFF) ranged from 0.38 to 6.51%. For the remaining product within this group (C/06, fillet blocks, fully cooked portions), the difference between method means was 21.73%. For the remaining 6 (16.67%) of 36 products, the DPFF mean was greater than the APFF mean. The differences between method means ranged from -0.03 to -2.76%. RSD values were considered acceptable (i.e., RDSr < 9% and RSDR < 12%) for all products studied. The modified method for determining FFC in frozen coated fish products has been adopted first action to replace AOAC Official Method 971.13. PMID- 9419865 TI - Validated method for quantitation and identification of 4,4-desmethylsterols and triterpene diols in plant oils by thin-layer chromatography-high resolution gas chromatography-mass spectrometry. AB - Alkaline hydrolysis was performed on a series of different vegetable oils. The unsaponifiable lipid matter was extracted with ethyl ether, and the class of 4,4 desmethylsterols (sterols) plus the triterpene diols (diols) erythrodiol, uvaol, and betulinol were isolated by thin-layer chromatography. A validated method using the acetate derivatives of sterols instead of their silyl ethers is presented. The acetate derivatives were analyzed by high resolution gas chromatography (HRGC). Retention time, precision, recovery studies, and absolute response factors were calculated for these esters, and GC/mass spectrometric structure of the assigned retention times was confirmed for the sterols and triterpene diols. PMID- 9419867 TI - Fully automated determination of pesticides in wine. AB - A fully automated solid-phase extraction gas chromatographic/mass spectrometric (SPE/GC/MS) method was developed for determination of pesticides in wine. All steps from aspiration of infiltrated wine to printout of the integrated chromatogram were performed without human interaction. A dedicated robot performed addition of internal standard, application of wine onto the SPE cartridge, elution of analytes, drying and concentrating of eluate, and passing of concentrate to the GC sampler. All steps were performed in standard liquid chromatography/GC vials, using a minimum of organic solvent. The method permits determination of 21 different pesticides. Individual detection limits were 0.005 0.01 mg/L. The regression coefficients relating to linearity were > 0.99; only 4,4-dichloro-benzphenone and dicofol showed lower coefficients. The recoveries for 17 pesticides ranged from 80 to 115%. PMID- 9419866 TI - Determination of lead in wine by graphite furnace atomic absorption spectrophotometry: interlaboratory study. AB - An interlaboratory study of a graphite furnace atomic absorption spectrophotometry (GFAAS) method for the determination of lead in wine was conducted. Seventeen laboratories from France, United States, and the United Kingdom, using a variety of GFAAS instruments, took part in the study. The method incorporated a novel matrix-matching procedure to minimize matrix effects between standards and samples. Six wine test materials were prepared and sent to participants as 12 blind duplicate or split level samples. There was good agreement between results obtained from participants and target values (24-279 micrograms/L) obtained with an inductively coupled plasma-mass spectrometry method. The precision of the GFAAS method was well within the range predicted by the Horwitz equation for the 6 test materials analyzed. Repeatability standard deviations ranged from 3 to 17%. Reproducibility standard deviations were in the range of 10 to 30%. The method is recommended for use for official purposes. PMID- 9419868 TI - Survey of organochlorine pesticide residues in milk in Hong Kong (1993-1995). AB - A survey was conducted from 1993 through 1995 to monitor organochlorine pesticides and their metabolite residues in milk available in local Hong Kong markets. Of 252 samples analyzed, including pasteurized milk, fresh milk, and raw milk, 42 contained organochloride pesticide residues at levels exceeding the Extraneous Maximum Residue Limits of the Codex Committee on Pesticide Residues. DDE and HCH isomer levels were substantially higher than those found in a 1984 1987 survey, probably because the source of cow's milk has shifted from local dairy industries to mainland China over the past decade. Although organochlorine pesticides such as DDT and HCH have been banned in China since 1983, residues of such compounds may still persist in the environment and cause contamination through the food chain. PMID- 9419869 TI - Statistical classification of seafood quality. AB - Discriminant function analysis (DFA) was used to classify the freshness quality of lean fish, fatty fish, and shrimp as either acceptable (Class 1), marginal (Class 2), or unacceptable (Class 3). Fresh and frozen survey samples were statistically classified following an initial precategorization using sensory, chemical, and microbiological indices as predictor variables. Computer elimination of nonsignificant predictor tests, p > 0.05, was used to optimize the test protocol. DFA correctly classified 98.5% of 67 preclassed lean fish samples (34 Class 1; 13 Class 2; 20 Class 3), 86.2% of 58 preclassed fatty fish samples (22 Class 1; 16 Class 2; 20 Class 3), and 98.7% of 79 preclassed shrimp samples (45 Class 1; 18 Class 2; 16 Class 3) by using all the quality indices. Computer selection of significant predictor indices at p < 0.05 yielded correct predicted classifications of 95.5, 81.0, and 97.5%, respectively. The number of tests required to effectively categorize quality were reduced from 15 to 3 for lean fish, from 13 to 3 for fatty fish, and from 11 to 6 for shrimp, with minimal losses in prediction accuracy and a substantial reduction in analysis time. PMID- 9419870 TI - Spectrophotometric determination of pyridoxine hydrochloride [correction of hydrochlorine] pharmaceutical preparations and foods. AB - An easy and sensitive spectrophotometric assay of pyridoxine is described. The procedure is based on formation of an azo dye by the reaction of pyridoxine with diazotized 2,4-dinitroaniline followed by the reaction of the dye with Hg2+ ions to form a stable complex with maximum absorbance at 545 nm. The system obeys Beer's law for 4-75 micrograms pyridoxine hydrochloride in an overall aqueous volume of 25 mL (correlation coefficient, 0.9998). On extraction into 5 mL butan 1-ol, the system obeys Beer's law in the range 0.8-15 micrograms pyridoxine hydrochloride at 545 nm. The color is stable for 60 min in both aqueous and organic phases (molar absorptivity, 3.7 x 10(4) L/mol.cm; coefficient of variation, 3.1%, n = 10). The pyridoxine contents of pharmaceutical preparations, a processed foodstuff, and 2 rice samples were determined by using the proposed method. Assay reliability was established by recovery studies and parallel determination using a reported method. PMID- 9419872 TI - Analytical procedures for the determination of organotin compounds in sediment and biota: a critical review. AB - Analytical procedures reported over the last 10 years for the determination of organotin compounds in sediment and biota have been critically reviewed in terms of sample handling, sensitivity, analytical cost, environmental acceptance, accuracy and precision. Critical steps in the analytical procedures are identified. Finally, research needs in extraction and determination are suggested. PMID- 9419871 TI - Improvements in amperometric detection of sulfite in food matrixes. AB - Sulfite is added to foods as an antimicrobial, antibrowning agent, or antioxidant. It also can occur naturally, and is often used in the production of beer and wine. For years the standard methodology for determination of sulfite in foods has been the Monier-Williams method, which is a combination of acid distillation and titration. Recently, AOAC adopted a chromatographic method based on a method developed by Kim and Kim for the determination of sulfite. The method combines ion exclusion chromatography with direct-current (DC) amperometric detection to provide more convenient and accurate quantitation of sulfite. However, fouling of the platinum working electrode results in a rapid decrease in method sensitivity. As a result, standards must be injected before and after every sample, and the electrode must be polished frequently to maintain adequate detection limits. Pulsed amperometric detection overcomes electrode fouling problems by repeatedly and continuously applying cleaning potentials to the working electrode. Using this technique, a reproducible electrode surface can be maintained, and injection-to-injection repeatability is greatly improved. A comparison of method performance for both DC and pulsed amperometric detection is presented. Also investigated was the stability of sulfite samples at varying pH, and in the presence or absence of a preservative. PMID- 9419873 TI - Temperature programming and gradient elution in reversed-phase chromatography with packed capillary columns. AB - The two major anisocratic elution modes were compared in reversed-phase chromatography with 180 microns I.D. fused-silica capillary columns packed with 6 microns Zorbax SB ODS-silica. By evaluating the retention factors of alkylbenzenes at acetonitrile concentrations varying from 60 to 80% (v/v) in the aqueous eluent and in the temperature range of 30-80 degrees C, it was found that a 5 degrees C change in column temperature and a 1% change in acetonitrile concentration have almost the same effect on retention. This is illustrated by the almost identical chromatograms of an alkylbenzene sample obtained by temperature programming and by gradient elution under the same conditions otherwise and by simulation of the trajectories of the eluent peaks moving down the column. The results suggest that in reversed-phase HPLC with packed capillary columns temperature programming offers an alternative to gradient elution in a relatively narrow range of the required elution strength. Thermodynamic data from isocratic chromatographic measurements were used to predict the retention times of alkylbenzenes in reversed-phase chromatography with temperature programming at different heating rates and column inlet pressures. Temperature programming was used to separate beta-lactoglobulins A and B by reversed-phase chromatography. It was also employed concomitantly with gradient elution to enhance the separation of a mixture of four standard proteins. The results indicate that temperature programming could serve as an adjunct to gradient elution by means of fine retention tuning to bring about or increase the resolution of closely related macromolecules. PMID- 9419874 TI - Analysis of organic and inorganic selenium anions by ion chromatography inductively coupled plasma atomic emission spectroscopy. AB - We report analysis of both inorganic and amino acid forms of selenium by ion chromatography with inductively coupled plasma atomic emission spectroscopic detection. Three chromatographic systems are compared; effects of representative sample matrices on the separations are investigated. We are unable to resolve selenate and seleno-cystine using the Dionex AS4A column. Elution of seleno cystine and seleno-cysteine is strongly suppressed in samples of bacterial cell extract matrix analyzed with the Dionex AS10 column; this interference is not observed with the Dionex AS11 column. Synthetic sea water sample matrix has little effect on analytical results. Quantitation parameters are reported. PMID- 9419875 TI - Rapid semi-quantitative estimation of N-nitrosodibutylamine and N nitrosodibenzylamine in smoked hams by solid-phase microextraction followed by gas chromatography-thermal energy analysis. AB - A solid-phase microextraction (SPME) analytical method has been developed for the determination of N-nitrosodi-n-butylamine (NDBA) and N-nitrosodibenzylamine (NDBzA) in hams that is based on: (a) isolation of the compounds by steam distillation, (b) SPME from the distillate headspace using a polyacrylate coated silica fibre and (c) determination by gas chromatography-thermal energy analyzer technique or confirmation by gas chromatography-mass spectrometry. Recoveries of both NDBA and NDBzA from hams spiked at 5 to 160 micrograms/kg levels ranged between 41 to 112%. The overall method is fast, sensitive (detection limits, 1 to 3 micrograms/kg), precise (within 10%) and fairly accurate (average recoveries 86% and 70%, respectively). The results obtained by this technique for seven ham samples agreed fairly well with those obtained by an existing method (r2 = 0.97). The new method is solventless, environmentally friendly and useful for rapid monitoring purposes. PMID- 9419876 TI - Capillary zone electrophoresis with fluid-impervious polymer tubing inside a fused-silica capillary. AB - A fused-silica capillary was used as a mold in the in situ formation of, and as a sheath for, a highly crosslinked poly(styrene-divinylbenzene) inner tubing having 3-5 microns wall thickness. This 'tube-in-the-tube' construction with the thin walled, fluid-impervious polymer inner tubing precluded contact between the aqueous buffer solution and the inner wall of the fused-silica capillary. As a result, this structure withstood long-term treatment with highly alkaline solutions without deterioration. In order to hydrophilize the inner surface of the polymer tube, a polyoxyethylene oligomer was grafted to its inner wall and subsequently crosslinked. The inner tube with such hydrophilic coating was also stable to hydrolytic attack by 1 M NaOH. Although the nonpolar polymeric inner surface generated electrosmotic flow as if it had some fixed negative charges, the flow velocity became almost negligibly small once it had been hydrophilized. As illustrated by electropherograms, the hydrophilization of the polymeric inner tube greatly facilitated the CZE of basic proteins in the pH range from 3 to 6 without the need for additives in the electrophoretic medium to mask the silanol groups at the surface of quartz capillaries. PMID- 9419877 TI - High resolution multichannel fluorescence detection for capillary electrophoresis. Application to multicomponent analysis. AB - A wavelength-resolved fluorescence detector for laser-induced fluorescence detection in capillary electrophoresis (CE) is described that uses a charge injection device (CID) array detector Post-column fluorescence detection occurs using a sheath flow cell. The limit of detection for fluorescein is 4.8 x 10(-11) M (29,000 molecules), the spectral resolution is 0.56 nm/pixel, and the spectrograph/CID monitors a 250 nm spectrum throughout the 250-875 nm range. Custom array readout, data manipulation and data processing methods are described to convert wavelength/spatial CID images into electropherograms. The application of the system to characterizing bilirubins in human serum is described, demonstrating the ability to match electrophoretic peaks to standards using spectral information. PMID- 9419878 TI - Perceptual strategies in the estimation of physical quantities by orangutans (Pongo pygmaeus). AB - The perceptual strategies used by 4 orangutans (2 subadults, 2 adults) when choosing the larger of 2 volumes in a Piagetian conservation task were investigated. Three possible perceptual strategies were investigated: (a) direct perceptual estimation of the container's content independent of its shape, (b) use of the spatial and temporal cues provided by the pouring of liquid from one container to another, and (c) ability to initially identify the larger volume and track it across transformations disregarding misleading perceptual cues. Results indicated that the direct perceptual estimation strategy was the best candidate to explain the orangutan's systematic choice of the larger of 2 quantities. PMID- 9419879 TI - Chimpanzee (Pan troglodytes) pointing: hand shapes, accuracy, and the role of eye gaze. AB - The manual pointing of 2 signing chimpanzees, Moja and Tatu, was examined in 2 experiments. Experiment 1 investigated eye-gaze direction, hand use, and hand shape while pointing. Both chimpanzees obtained the attention of a human before pointing toward an unreachable object. During 100 trials, the chimpanzees alternated their eye gaze between the object and the human while pointing. Moja's points were left-hand biased, and Tatu showed no lateral hand bias. Both indexical and whole hand points were recorded. Experiment 2 tested the chimpanzees' ability to point accurately toward objects in close proximity to each other. Humans were able to reliably determine the locations toward which the chimpanzees pointed. Both chimpanzees showed left-hand biases, and a higher proportion of indexical points were observed than in Experiment 1. These results are compared and contrasted with recent hypotheses pertaining to the topography of chimpanzee pointing and the role of eye gaze in deictic interactions. PMID- 9419880 TI - Pointing, withholding information, and deception in capuchin monkeys (Cebus apella). AB - Brown capuchin monkeys, like 4-year-old children and human-socialized chimpanzees, showed communicative and deceptive pointing in experiments in which they benefited by indicating, accurately or falsely, the location of hidden food. All 3 capuchin monkeys tested (13, 19, and 26 years old) pointed communicatively in the presence of a cooperative trainer. One human-reared monkey pointed without any training and frequently gazed at her human respondent; as with apes, extensive exposure to humans may promote some human-like responses in monkeys. Another capuchin withheld pointing when beneficial, whereas the 3rd learned to obtain the hidden food by pointing deceptively in the presence of a competitive trainer. Such deceptive pointing by one monkey and withholding of information by another suggest that primates' deceptive pointing in an experimental situation is explainable in terms of response inhibition and conditional discrimination learning. PMID- 9419881 TI - Maternal presence and rearing condition affect responses to a live predator in kangaroo rats (Dipodomys heermanni arenae). AB - Experiment 1 compared the responses of wild-caught adult and captive-born adult and juvenile kangaroo rats (Dipodomys heermanni arenae) to a live snake. Wild caught adult rats were less active and monitored the snake more than during a control condition; captive-born juvenile rats did not behave differently during snake and control tests. Snake-naive adult rats behaved more like the wild-caught adult rats, but not on all measures. In Experiment 2, pups were tested at 25 and 50 days of age in 4 conditions: no-snake control, alone with the snake, with a sibling and the snake, and with the mother and the snake. Pups did not behave differently during control and snake tests, but during tests with the mother, pups faced the snake less and followed the mother. Younger pups were more often near the mother than a sibling and followed the mother more when the snake was present. Development of defensive behavior may depend on both predator experience and maternal influence. PMID- 9419882 TI - Food transfers through mesh in brown capuchins. AB - Capuchin monkeys (Cebus apella) share food even if their partner is behind a mesh restraint. Pairs of adult capuchins were moved into a test chamber in which 1 monkey received cucumber pieces for 20 min and the other received apple slices during the following 20 min. Tolerant transfers of food occurred reciprocally among females: The rate of transfer from Female B to A in the second test phase varied with the rate from Female A to B in the first test phase. Several social mechanisms may explain this reciprocity. Whereas this study does not contradict cognitively complex explanations (e.g., mental record keeping of given and received food), the results are consistent with a rather simple explanation: that food sharing reflects a combination of affiliative tendency and high tolerance. The study suggests that sharing mechanisms may be different for adult male capuchins, with males sharing food more readily and less discriminatingly than females. PMID- 9419883 TI - Effects of social interaction on the development of starling song and the perception of these effects by conspecifics. AB - To examine the effects of contact with a conspecific in the absence of species typical song models, the authors raised starlings in male-male pairs in acoustic isolation. The songs of these birds differed significantly from those of either individual isolates or wild adults and resembled in some respects the songs of starlings tutored by live conspecifics. Operant conditioning techniques were used to demonstrate that these differences among songs were perceptually salient to conspecifics. The results indicated that (a) wild-caught adult starlings are capable of forming open-ended categories for isolate and wild song, (b) starlings perceive the songs of isolated pairs as more "isolatelike" than "wildlike," and (c) starlings can distinguish the songs of isolated pairs from those of individual isolates. Both experiments point to the importance of social factors in avian song development. PMID- 9419884 TI - Interaction with demonstrator rats changes observer rats' affective responses to flavors. AB - The authors examined whether exposing naive rats (observers) to recently fed conspecific demonstrator rats changed the observers' later affective responses to foods their demonstrators ate. In Experiment 1, observers learned an aversion to a flavored fluid, then interacted with demonstrators that had drunk that fluid. These observers, but not those interacting with demonstrators that had drunk water, increased their intake of the averted fluid and exhibited fewer negative responses when the averted fluid was infused into their mouths. Rats in Experiment 2 entered the arm of a T maze known to lead to banana-flavored pellets more frequently after interacting with demonstrators fed banana-flavored pellets than after interacting with demonstrators fed chow-flavored pellets. Results of both experiments indicated that interaction with demonstrator rats changed observer rats' affective responses to flavors. PMID- 9419885 TI - Individual differences and their relation to social structure in domestic cats. AB - Two groups of domesticated cats with established social structures were tested for their response to novel stimuli in a variety of test situations. Clear individual differences in responses were consistent over a series of test sessions and remained stable despite regular environmental changes. Individual differences in behavioral response were not related to object dominance (food competition) or social dominance (freedom of movement in social encounters) unlike similar studies with social animals. No relationship was found in either group between a cat's rank in object dominance and its rank in social dominance. Significant correlations were demonstrated in the individual cat's latency to approach a novel stimulus, behavioral rank in test situations, and attention span in both groups of cats. Comparisons were made between similar studies with wolves, and inferences were drawn about the relationship between individual differences and social structure in social and nonsocial species. PMID- 9419886 TI - Effects of the nonagouti coat-color allele on behavior of deer mice (Peromyscus maniculatus): a comparison with Norway rats (Rattus norvegicus). AB - The agouti locus influences coat color by antagonizing melanocyte-stimulating hormone (MSH) at its receptor on pigment cells and may antagonize MSH in neural tissue. This study replicates work on rats to assess whether behavioral (neural) effects of the agouti locus are as similar across mammals as those on coat color. Handling, open-field, platform jump, and food-novelty tests were conducted on agouti and nonagouti deer mice (Peromyscus maniculatus) following protocols in C. A. Cottle and E. O. Price (1987). As with rats, nonagouti deer mice were less aggressive, less active, and easier to handle compared with their agouti counterparts. Nonagouti deer mice also groomed more than agouti subjects. Thus, behavioral effects of the agouti locus are conservative, and agouti may be an important modulator of melanocortins in neural as well as integumentary tissue. PMID- 9419887 TI - Linkage map for the Asian tiger mosquito [Aedes (Stegomyia) albopictus] based on SSCP analysis of RAPD markers. AB - A linkage map of the Asian tiger mosquito [Aedes (Stegomyia) albopictus (Skuse)] was constructed in an F1 intercross by monitoring the segregation of randomly amplified polymorphic DNA (RAPD) markers analyzed for single-strand conformation polymorphisms (SSCP). We hypothesized that SSCP analysis would reveal point mutations in RAPD fragments that would then segregate as codominant rather than dominant markers which are typically revealed through routine RAPD analysis. Markers were mapped to individual chromosomes by testing for cosegregation with Sex (chromosome I) or a polymorphism at the a-GPD allozyme locus (chromosome II). All other markers that cosegregated were assigned to chromosome III. Six RAPD primers amplified 68 polymorphic markers that segregated in a Mendelian fashion and were mapped. Contrary to our hypothesis, no codominant SSCP polymorphisms were detected, but fractionation of RAPD products on polyacrylamide gels and detection through silver staining proved to be a sensitive technique that allowed us to identify more markers than the standard analysis of RAPD PCR products on agarose gels. PMID- 9419888 TI - Genetic analysis of triploidy in a selected line of chickens. AB - We investigated the pattern of inheritance of maternal meiotic errors responsible for a high frequency of triploid progeny in a selected line of chickens. For the genetic analysis, F1 and backcross populations were produced from crosses between normal diploid individuals of the triploidy line and a control line. Triploid embryos were produced by 35% and 67% of reciprocal F1 females and by 24% and 67% of reciprocal backcross females. These results exclude autosomal recessive and sex-linked recessive or sex-linked dominant inheritance. A single autosomal dominant gene is also not likely to be responsible. However, the results are consistent with the determination of triploidy by a single autosomal gene with no dominance, and an even better fit is obtained by two loci, an autosomal gene with no dominance and a sex-linked gene. The results cannot exclude a multifactorial mode of inheritance, but the rapid response to selection for triploidy and consistent expression of the meiotic errors in different genotypes suggest that meiotic mutations at one or two loci are the most plausible genetic basis for the trait. PMID- 9419890 TI - Identification of Rfp-Y (Mhc-like) haplotypes in chickens of Cornell lines N and P. AB - Two strains of chickens selected for differential Marek's disease (MD) resistance or susceptibility were studied for the presence of the recently described Rfp-Y major histocompatibility complex (Mhc-like) haplotypes. MD resistant chickens from line N were fixed for the classical Mhc B21 haplotype, whereas MD susceptible line P chickens were fixed for the B19 haplotype. The Rfp-Y haplotypes were identified by restriction fragment polymorphism (RFP) analysis using enzymes and Mhc probes for B-G, B-L beta II, and B-FIV. In addition an Rfp L beta III clone was developed that differentiated Rfp-L beta from B-L beta genes. Three Rfp-Y haplotypes, defined for both class I and class II Mhc-like loci, were identified in line N (Rfp-Y5, Rfp-Y7, and Rfp-Y8) and in line P (Rfp Y5, Rfp-Y8, and Rfp-Y9), respectively. The Rfp-Y7, Rfp-Y8, and Rfp-Y9 haplotypes have not been described previously. The Rfp-Y5 haplotype was most frequent (0.70) in line N, but existed in low frequency (approximately 0.04) in line P; the Rfp Y9 haplotype was most frequent in line P (0.63), but was absent in line N. The Rfp-Y haplotypes-segregated in a Mendelian fashion in each line based on analysis of progeny from Rfp-Y heterozygous matings. The frequency of recombination between the Rfp-Y F and L loci was estimated to be less than 0.25%. PMID- 9419889 TI - A comparative analysis of Mhc DRB3 polymorphism in the American bison (Bison bison). AB - The degree of genetic polymorphism at the DRB3 locus in the major histocompatibility complex (Mhc) of the North American bison was investigated by PCR and DNA sequence analysis. Nine different alleles were characterized in a selected sample of 20 animals. The genetic distances between alleles were as large as usually found at highly polymorphic Mhc loci in other species. A comparative analysis of the DRB3 polymorphism in bison and cattle revealed an extensive sharing of sequence motifs. The result clearly shows a transspecies persistence of DRB3 allelic lineages in the two species. Consequently a significant amount of Mhc polymorphism has been maintained through the population bottleneck that bison experienced in the late nineteenth century. An analysis of the pattern of sequence polymorphism among bison and cattle DRB3 alleles strongly suggested that interallelic recombination has contributed significantly to the generation of allelic diversity at this locus. PMID- 9419891 TI - X-linked ectodermal dysplasia in the dog. AB - A male German shepherd pup had symmetrical areas of hairlessness as well as missing and misshapen teeth. There was no family history of a similar phenotype. In biopsies of the hairless skin and foot pads there were no hair follicles, adnexal structures, or eccrine glands. These findings resemble those in ectodermal dysplasia in the Tabby mouse and anhidrotic/hypohidrotic ectodermal dysplasia (HED) in man, which are both X-linked recessive disorders and thought to be homologous gene defects. While similar cases of ectodermal dysplasia have been reported in the dog and some genetic studies carried out, definitive confirmation of X-linked inheritance of canine ectodermal dysplasia is lacking. Family studies and experimental matings using the propositus gave results that confirm X-linked recessive inheritance. On statistical grounds, it is concluded that ED in the propositus is due to a new mutation. A colony of dogs with this mutation is maintained for further study. PMID- 9419892 TI - Microsatellite variation in an introduced mouflon population. AB - As previous studies of genetic polymorphism in the mouflon (Ovis gmelini) have not provided any valuable markers for population studies, we tested the capacity of microsatellites to index the genetic diversity of a recently introduced mouflon population. Six pairs of bovine primer amplified microsatellites in mouflon, and all six were polymorphic. Furthermore, despite the low number of founders, five loci had a high gene diversity in this introduced population. Unlike other genetic markers, microsatellites could be powerful to study the genetic structure of mouflon populations. PMID- 9419893 TI - Molecular mapping of insecticide resistance genes in the yellow fever mosquito (Aedes aegypti). AB - Several loci conferring insecticide resistance in the yellow fever mosquito (Aedes aegypti) have previously been mapped by simple recombinational mapping. Here we describe correlation of these resistance phenotypes with molecular gene probes for insecticide target sites by RFLP mapping. The para sodium channel gene homologue and the GABA receptor gene Resistance to dieldrin map to the same genome regions as the DDT/pyrethroid and cyclodiene resistance loci, respectively. Although the acetylcholinesterase (target site of organophosphorus and carbamate insecticides) gene Ace does not map to any known resistance locus, it maps very close to the sex-determining locus. We discuss the possibilities that, if identified, Ace-mediated resistance in A. aegypti will be sex linked or that, as suggested for anopheline mosquitoes, two independent Ace loci may exist, one of which is autosomal. These results support the importance of target site insensitivity as an insecticide resistance mechanism in mosquitoes. PMID- 9419895 TI - Sex linkage of minisatellite bands in bobcats (Felis rufus). AB - Minisatellite DNA profiles using the multilocus human probe 33.6 are presented for 27 captive bobcats (Felis rufus) of documented geographic and genetic origins. The results show that 30% of the finger-printing bands present in males are sex linked. The effect of sex on band sharing was attributed to the presence of male-specific hemizygotic bands belonging to a minisatellite cluster located in the nonrecombinant region of the Y chromosome. A combination of mechanisms might drive the dynamics of minisatellite loci and allow different evolutionary rates depending on the recombinational capability of the chromosomal locations involved. We discuss the utility of sex-linked fingerprinting bands as genetic markers for the study and management of bobcats. PMID- 9419894 TI - Comparative mapping of anchor loci from HSA19 to cattle chromosomes 7 and 18. AB - Six loci--CALR, EPOR, JUNB, JUND, CEA, and PRKCG--were assigned to bovine chromosomes using PCR-based hybrid somatic cell analysis. The five genes other than CALR are comparative mapping anchor loci. This study, together with the previous assignment of three anchor loci--INSR, LDLR, APOE--and four other genes- AMH, GPI, RYR1, LHB--defines the conserved synteny relationship between human chromosome 19 and cattle chromosomes 7 and 18. Genes on HSA 19p13.3-13.2 are conserved in cattle chromosome 7, while those on HSA19-q13.1-13.4 are conserved in cattle chromosome 18. In contrast, homologous genes from HSA19 are located on four different mouse chromosomes, namely MMU10, MMU8, MMU9, and MMU7. This is further evidence that syntenic conservation between cattle and human generally exceeds that observed between human and mouse. PMID- 9419896 TI - Karyotypic analysis of C-banded chromosomes of diploid alfalfa: Medicago sativa ssp. caerulea and ssp. falcata and their hybrid. AB - Chromosomes of two diploid (2n = 2x = 16) subspecies of Medicago sativa ssp. caerulea and ssp. falcata and their hybrid were studied by C-banding. This study was undertaken to improve the C-banding technique for alfalfa chromosomes, develop a C-banded karyotype of the ssp. caerulea and ssp. falcata, and determine if the same C-banding technique could be used to identify parental chromosomes in hybrids. The chromosomes of ssp. falcata have only centromeric bands and thus individual chromosomes could not be identified. One accession of ssp. falcata displayed an interstitial band in the middle of the long arm on the satellite chromosome. However, chromosome-specific bands were observed in ssp. caerulea enabling the identification of each of the eight pairs of chromosomes and the development of a idiogram. All chromosomes had centromeric bands and a terminal band in the short arm except the satellite chromosome (chromosome 8). Interstitial bands were also observed in the short arms, with the exception of chromosome 7. Chromosomes 1, 2, 3, and 8 each had one prominent interstitial band in their long arm. The satellited chromosome is easy to identify because of the presence of the secondary constriction, two bands located on either side of the nucleolar organizer region, and a large terminal band on its long arm. The differences in banding patterns between these subspecies allowed the identification of parental chromosomes in hybrid cells. PMID- 9419897 TI - Differences in pollen-tube growth rate and reproductive isolation between Louisiana irises. AB - A level of reproductive isolation is necessary for the process of genetic divergence. Such isolation also prevents the homogenization of species following secondary contact. This study is an investigation of the relative contribution of two prefertilization mechanisms, pollen-tube growth rate and pollen-tube attrition, to the reproductive isolation of Iris fulva and I. brevicaulis, two naturally hybridizing members of the Louisiana iris species complex. Flowers of each species were first pollinated with heterospecific pollen. After various time intervals, conspecific pollen was added. Analyses of the patterns of resulting progeny were used to infer whether relative pollen-tube growth rates act as a prefertilization isolating mechanism. In I. fulva the frequency of hybrid seeds increased with increasing pollination interval, suggesting that hybridization is limited by pollen-tube growth rates. Likewise, in I. brevicaulis hybrid sed production increased, but it was high regardless of the pollination interval. Thus it appears that relative pollen-tube growth rates limit interspecific reproduction in both species, but barriers are weaker in I. brevicaulis. PMID- 9419899 TI - Fructose 1,6-bisphosphate aldolase activity in leaves of a rice mutant selected for enhanced lysine. AB - Unknown proteins isolated from mutant tissues of rice (Oryza sativa L.) recovered from inhibitor selections were subsequently peptide microsequenced. Database searches putatively identified one peptide as fructose 1,6-bisphosphate aldolase (EC 4.1.2.13). Tissues of mutant rice, PI564784, and wild type (cv Calrose 76) tissues were evaluated for aldolase activity. Total enzyme activities were slightly lower in the mutant than the control but the differences were not significant. Although the mutant phenotype is for enhanced lysine and protein, we ascribe the small aldolase differences to physiological adjustments, rather than to DNA modifications of the aldolase gene(s). Homologies of rice peptides with aldolases from a range of species, as well as rice cell culture expressed sequence tags (ESTs) are presented. Some amino acids sequences are highly conserved. The mutant phenotype expressing stress proteins is not likely to be defined by a change in rice aldolases. PMID- 9419898 TI - Carica papaya latex is a rich source of a class II chitinase. AB - A class II chitinase is present in the latex of the tropical species Carica papaya. The enzyme may be readily purified by using a combination of hydrophobic interaction- and cation-exchange chromatography. This enzyme preparation is homogeneous with respect to the three physico-chemical criteria of charge, M(r) (28,000) and hydrophobicity. It is also completely free of any proteolytic and bacteriolytic activities. The enzyme was classified as a class II chitinase on the basis of its N-terminal amino acid sequence up to the 30th residue. In agreement with this classification, the enzyme preparation hydrolyses chitinase substrates only very slowly and several free thiol functions are present in the polypeptide chain. These free thiol functions are buried, and to be available for titration with 2,2'-dipyridyldisulphide, the enzyme must be denatured. Unfolding of papaya chitinase requires particularly drastic conditions, not less than 4 M guanidinium hydrochloride at 25 degrees and pH 6.8. PMID- 9419900 TI - Chrysin and other leaf exudate flavonoids in the genus Pelargonium. AB - In a chemotaxonomic survey of 57 Pelargonium species, leaf exudate flavonoids were detected in 35% of the sample, mostly in trace amounts. However, chrysin and a related C-methylflavanone were identified as major leaf surface constituents of P. crispum, and a mixture of quercetin and kaempferol mono-, and di- and trimethyl ethers of P. quercifolium. In two other species, P. fulgidum and P. exstipulatum, methylated flavones were the only lipophilic flavonoids present. This is the first report of leaf surface flavonoids from the genus Pelargonium. PMID- 9419901 TI - The doctor's dilemma: defining worth. PMID- 9419902 TI - Just say no. PMID- 9419903 TI - In the interim. PMID- 9419904 TI - Cleaning house. PMID- 9419905 TI - TB hot spot. PMID- 9419906 TI - Learning in Laredo. PMID- 9419907 TI - Histopathology, clinical findings and treatment of renal hydatidosis. AB - Renal hydatidosis, although not frequent, is the third location of the echinococcus granulosus in the man, after the liver and lungs. Its clinical importance is remarkable as the natural history of the process (if left untreated) usually results in the opening of the parasitic focus into the excretory ducts and the complete destruction of the kidney. In spite of that, insufficient attention is usually dedicated to the topic. So in the lecture the major aspects of the problem have been considered, also on the basis of histological, clinical and radiological personal studies. The site localisation is essential, and at present is possible on combined modality studies, in particular on diagnostic imaging: plain radiography, conventional urography and new imaging procedures, US, CT and MR, give typical images that definitively prove the specific diagnosis. With regard to treatment, the problem at present is still basically surgical: but nephrectomy (as excessively performed in the past) must be if possible avoided and reserved for cases of parenchymal destruction or of untreatable infection). On the contrary, when it is still possible, one must consider conservative techniques, limited to the removal of the parasite and of permanently damaged parenchymal areas. Cystectomy with resection of the exuberant pericystium or (when possible) segmental resection of the kidney offer a good solution, sometimes also in cases of recurrence. PMID- 9419908 TI - Pancreatic cancer and tumor markers: role of the newly identified CAR-3 antigen. AB - BACKGROUND AND AIM: The aim of this study is to assess the clinical usefulness of the serum assay for CAR-3 in the diagnosis and follow-up of pancreatic cancer. MATERIALS AND METHODS: Serum levels of tumor markers (CAR-3, Ca 19.9, Ca 195 and CEA) were measured in a total of 238 patients with various diseases of the gastrointestinal (GI) tract, including 61 pancreatic cancers. Cut-off levels were calculated on the basis of a non-parametric estimate of 90% specificity. After surgery, patients with pancreatic cancer underwent a combined serological and radiological (CT-scan) follow-up. RESULTS: At the cut-off level of 6.15 U/L, the sensitivity of CAR-3 was 62.3% (CA 19.9: 77%; Ca 195: 75.4%; CEA: 24.5%). In the differential diagnosis between pancreatic cancer and other GI diseases, significant differences were found. No association was discovered either between serum level of tumor markers and tumor stage or between short- and long-term survivors. In the follow-up, CT-scan was superior to serologic tests (sensitivity: 94.2%). Among tumor markers, CAR-3 achieved a sensitivity of 62.5% (Ca 19.9: 83.3%; Ca 195: 75%). DISCUSSION: CAR-3 is shed in the circulating stream in a much lower proportion of cases than that observed for antigen expression at immunohistochemistry. During the follow-up CT-scan was the most accurate diagnostic tool. However, the meagre therapeutical options for recurrent pancreatic cancer, do not justify such an aggressive follow-up. CONCLUSIONS: Ca 19.9 remains the tumor marker of choice for either the pre-operative work-up or the post-surgical follow-up of patients with pancreatic cancer. PMID- 9419909 TI - Acute pseudo-obstruction of the colon (Ogilvie's syndrome): advances in management. AB - The management of the patients with acute pseudo-obstruction of the colon (APOC) still represents a matter of debate. To better evaluate and compare the effectiveness of various therapeutic approaches in the management of APOC 29 patients were considered. These were included according to three consecutive periods in: group A (1977-1982) concerning patients who underwent medical treatment alone (n = 8) or endoscopic (n = 4) and surgical (n = 1) decompression; group B (1983-1990) in which the management was based on simple endoscopic decompression (n = 10); group C (1991-1995) including patients in whom placement, under fluoroscopic control, of a tube in the cecum following endoscopic decompression was provided (n = 6). Mean time required for resolution of colonic distension was 2.3 (+/- 0.50 SD) days in patients who underwent endoscopic decompression and tube placement, as compared to 4.5 (+/- 2.47 SD) days in the group of patients treated either with conservative measures or simple endoscopic decompression (p = 0.04). No recurrence occurred after colonoscopic decompression and tube placement while colonic distension recurred in 4 of 14 patients managed by simple endoscopic decompression (0% vs. 28.6%, n.s.). Our experience showed that endoscopic decompression is an effective method, moreover if associated with the placement of an indwelling tube into the right colon. This method, for its easiness and safeness, besides its effectiveness in preventing the recurrence of colonic distension, may be surely considered an advance in the management of acute pseudo-obstruction of the colon. PMID- 9419910 TI - Tension-free hernioplasty: technical remarks and personal experience. AB - The authors report their experience with 463 tension free hernioplasty procedures for inguinal and femoral hernias. The surgical technique included the insertion of both a polypropylene plug and a polypropylene mesh, and was carried out mostly in local anesthesia (84.2%) using bupivacaine 0.25% for ileoinguinal and ileohypogastric blockage and mepivacaine 0.5% for local infiltration. There was no major intraoperative complication; local postoperative complications were rare (10%) and easily managed; postoperative pain was frequently observed (66%), though mild and transient; resumption of working activity occurred within a month in 96.6% of cases; there were only 3 post-operative recurrences (respectively, at 1, 6 and 12 months). The authors conclude that the tension free hernioplasty is a simple, rapid, low-cost and effective technique, easily performed under local anesthesia. PMID- 9419911 TI - The power of politics and partnerships. PMID- 9419912 TI - Ameliorating adults' acute pain during phlebotomy with a distraction intervention. AB - This study evaluated the effectiveness of a distraction intervention on subjects' perceptions of pain. During phlebotomy, 96 adults received either usual care or used a kaleidoscope as a distraction. After phlebotomy they rated their level of experienced pain with each of three instruments: Wong-Baker FACES Pain Scale, pain visual analogue scale, and Present Pain Intensity Scale. Statistical analyses revealed significantly lower perceptions of experienced pain among subjects using the kaleidoscope and concurrent validity for using the FACES Pain Scale with adults. Because the distraction intervention is effective, inexpensive, and easy to implement, its routine use during phlebotomy is recommended. PMID- 9419914 TI - Tattooing: another adolescent risk behavior warranting health education. AB - A cross-sectional, convenient sample of adolescents (N = 2101) from 8 states were queried regarding interest in tattooing. Permanent markings and blood-borne diseases were reasons respondents refrain from tattooing, yet 55% (n = 1159) expressed an interest in tattooing. Tattooed adolescents in the sample (10%, n = 213) responded with their experiences. Tattooing was frequently done around the 9th grade and as early as 8 years of age; over half (56%, n = 120) report academic grades of As and Bs. Potential health risks and definite psychosocial findings of purchase and possession risks were evident, building on data from a similar 1994 study by Armstrong and McConnell. Health providers and educators should initiate applicable health education and become community adolescent advocates regarding this risk-taking behavior. Findings indicate that adolescents who want a tattoo will obtain one, regardless of money, regulations, or risks. Adolescents view the tattoos as objects of self-identity and body art, whereas adults perceive the markings as deviant behavior. Informed decision-making could be promoted in health education by incorporating information about the possibility of blood-borne diseases, permanent markings, and themselves as growing and changing people. PMID- 9419913 TI - The influence of ostomy surgery on body image in patients with cancer. AB - A one group repeated measures design was used to describe changes in body image in 45 patients treated with ostomy for bowel or bladder cancer. Body image was measured preoperatively and at two postoperative points: 1 month and in 6 months or postclosure. The Body Cathexis Scale, Draw-a-Person, and subjective responses to open-ended questions were used. There was a significant difference in Body Cathexis scores between the immediate 1 month postoperative period and 6 months later (F(2, 88) = 3.13, p = .049). Body image scores did not change significantly between the preostomy and first postostomy measures. Qualitative data suggested that cancer diagnosis and its associated concerns were paramount on subjects' minds preoperatively and thus negatively influenced body image before the creation of the ostomy. Although altered body image is an appropriate nursing diagnosis for patients with ostomy in both the preoperative and postoperative periods, other issues may be dominant especially in the preoperative period. PMID- 9419915 TI - The effect of developmental care on preterm infant outcome. AB - Using a retrospective comparative design, the investigators evaluated the effects of training 10% of a nursing staff in the Neonatal Individualized Developmental Care and Assessment Program (NIDCAP) on preterm infant outcomes. A convenience sample of 25 preterm infants (< 1500 grams) cared for during the NIDCAP implementation and training period were matched by birth weight and gestational age to infants born before NIDCAP implementation. Outcome measures collected through chart review showed NIDCAP infants had significantly fewer and less severe intraventricular hemorrhages, fewer days of ventilatory support, shorter hospitalizations, and greater rate of weight gain. Findings suggest that benefits of developmental care are achievable with only a portion of the staff being NIDCAP trained. PMID- 9419916 TI - Nocturia and nocturnal urine production in obstructive sleep apnea. PMID- 9419917 TI - The development and pilot testing of The Diabetes Activities Questionnaire (TDAQ): an instrument to measure adherence to the diabetes regimen. PMID- 9419918 TI - Self-performance survey analysis. PMID- 9419919 TI - New research questions in nursing management. PMID- 9419920 TI - Managing emotions and success in life. PMID- 9419921 TI - Risk profiles for institutionalization in a cohort of elderly people with dementia or depression. AB - Seventy-five elderly persons with dementia or depression, served by a nursing outreach assessment and case management service in Belgium, were entered in a risk profiling study. Cluster analysis yielded three clusters, each presenting a different risk profile for institutionalization: (1) High Risk Profile, with subjects of moderately advanced age, highly dependent for activities of daily living (ADL), with severe cognitive impairment, poor communication skills, and behavioral problems; (2) Moderate Risk Profile, with subjects of advanced age, limited ADL-dependency, yet high care demands for Instrumental ADL (IADL), moderate to severe cognitive impairment, adequate communication competency, and some behavioral problems; and (3) Low Risk Profile, consisting of relatively young elderly, partially ADL and IADL-dependent, mild or no cognitive impairment, good communication abilities, and no particular behavioral problems. PMID- 9419922 TI - Psychobiology and psychopharmacotherapy of unipolar major depression: a review. AB - Psychopharmacology has continually informed the biological perspective of major depression. Antidepressants affect a variety of neurotransmitters by a wide range of pharmacological actions. The complexity of these neurotransmitter receptor interactions likely underlies the discrepancies observed in biochemical and physiological responses among apparently clinically homogenous depressive subgroups. This report provides an integrated review of the neuroendocrine and neurochemical perspectives of unipolar depression and how these advances influence the psychopharmacotherapy of unipolar major depression. PMID- 9419923 TI - The lifeworld of the chronic mentally ill: analysis of 40 written personal accounts. AB - In this study, chronicity in mental illness has been investigated as it is lived rather than how it might be conceptualized. Published first-hand accounts have provided the mechanism for direct access in coming to know the life of persons, their circumstances, and the meanings they associate with a life of persistent and enduring mental illness. These are unique and particular human experiences, and there are no empirical generalizations or law-like statements that can give such an understanding. Therefore the disclosure of meaning was sought through a hermeneutic-phenomenologic process. Four lifeworld existentials provide the framework for a combined description and interpretation of what it means to "live" chronic mental illness. The article concludes with a brief discussion of some implications for nursing practice, and commentary is made on the relevance of such insights to health care providers in both acute and community care settings. PMID- 9419924 TI - Deconstructing progress notes in psychiatric settings. AB - The purpose of this article is to examine the language by which nursing staff described these patients in their medical records. Charts of former psychiatric patients were studied for the quality of the words used to communicate about patient behaviors. More than 4,000 entries were examined and descriptive words that appeared with most regularity were presented to a panel of expert psychiatric nurses who were asked to Q sort them as to connotation. Results show that nurses entries focus on patients in judgemental and unflattering ways. Recommendations are made to replace the use of negatively laden language and labels with descriptions in more behavioral and operational terms. PMID- 9419925 TI - Evolution of community mental health case management: considerations for clinical practice. AB - Case management has been the preferred method of helping persons with mental illness live in the community. This report examines the historical development of case management practice over the past 30 years. The analysis revealed that evolutionary growth in case management developed in response to expansion of community mental health services rather than consumer interests to improve their quality of life. Consequently, this reveals itself in the lack of outcome criteria needed to measure case management effectiveness. Currently, the concept of case management is undergoing a revolutionary paradigm shift that is "consumer driven" and responds to consumer concerns. However, as mental health care costs rise within a rapidly growing community mental health care system, managed care is being implemented as a method to contain these rising costs. Because of major differences in philosophical underpinnings, developments in support of public mental health managed care may jeopardize the genesis of consumer-driven case management by supporting case management models that reflect managed care ideology. PMID- 9419926 TI - Voices from practice: mental health nurses identify research priorities. AB - Through a collaborative research project, nurse clinicians and nurse academics identified and prioritized the most important research questions arising from the current and future practice of mental health nurses in an eastern Canadian region. Study methods included the Delphi technique and a nominal group survey approach. Nine categories of research questions emerged. The top four categories are discussed under the headings: Preparation of Helpers, the Service System, Caregiver Needs, and Clients With Major Behavior Problems. This timely study offers explicit direction for psychiatric and mental health nursing practice and research, and for the development of social policy. PMID- 9419927 TI - A strategic supportive model for health prevention in the elderly: profile of a Puerto Rican geriatric population in a public health sector. AB - OBJECTIVE: To introduce the ASSUME study with the presentation of a clinical, socio-demographic, preventive and psychological profile of a geriatric population of patients who receive their health care in the General Internal Medicine Ambulatory Sector of our institution. METHODS: The Assume study is a prospective, randomized trial which is directed at increasing the participation of patients in preventive health care strategies at a primary, secondary and tertiary level. In this paper we focus on the initial stage of the process which aims to define and synthesize predisposing risk factors in the geriatric patient which would be amenable to primary, secondary and tertiary preventive strategies. Through a process of patient interview profiles of a physical, social and psychological nature are have been constructed. With the availability of this profile a clearer definition of the potential benefit of preventive strategies could be established. In this paper we present the initial profile of patients of all patients randomized to the study as of Sept. 01, 1997. RESULTS: A total of 123 patients have been enrolled with 48(39%) males and 75(61%) females. The mean age of patients is 70 years with a median of 68 years. Cardiovascular disorders establish the leading disease events in our population of patients with Hypertension in 85%, Ischemic heart disease in 50%, Myocardial Infarction in 19% and 40% with a history of Congestive Heart failure. Diabetes and Heart Failure were seen in 40%. An average of 4.4 prescribed drugs per patient was documented. A minority of patients took more than 7 drugs and none took more than 9 medications. Most patients (67 or 55%) had not required hospitalizations in the preceding 12 months and none of the patients required more than 4 hospitalizations. The average LOS was 8.60 days. The Preventive Medicine profile reveals a large number of un-vaccinated adults. Regular cigarette smoking was seen in 12%. We have used the body mass index as a measure of adequacy of weight. We highlight the number of patients who have a BMI equivalent to an obese, severely obese or morbidly obese category (41%). The number of patients who follow a prescribed diet was found to be 54 patients for 44% of the study group. With regards to the interventions primarily designed for early cancer detecting, approximately half of the patients undergo the recommended annual screening interventions. The screening of visual accuracy was reported in 54%, dental screen (24%) and auditory screening (15%). Nearly a quarter of patients have severe depression. The CAPE testing reveal that in the information and orientation section most patients presented none or light dysfunction(87%). In the conductual phase marked or severe impairment was detected in 12% of patients. In the mental ability section 22% of patients presented marked or severe impairment. CONCLUSION: The geriatric population studied would benefit from modalities which would increment the modern modalities for primary and secondary prevention of disease. Follow-up studies will allow the evaluation of the effectiveness of the conceptual model proposed, which would increment the patient participation in these preventive modalities. PMID- 9419928 TI - Outline of the Human Retrovirus Registry: profile of a Puerto Rican HIV infected population. AB - OBJECTIVE: To present the general socio-demographic profile, some risk related parameters and elements of the clinical spectrum of disease at presentation, of those HIV/AIDS patients enrolled in the Human Retrovirus Registry. METHODS: This is a prospective longitudinal cohort study, which has been identifying since May 1992, adults or adolescents 18 years or older with AIDS or HIV infection at the time they present to our health care facilities: University Hospital Ramon Ruiz Arnau and the Bayamon Immunology Clinic. The present analysis include patients enrolled between May 1992 and December 1996 (n = 1520). The measurement instrument is a modular questionnaire which actually includes 237 variables including socio-demographic data, risk variables, lifestyle and affective parameters, clinical and immunological variables and therapeutic data. RESULTS: The mean baseline age of the 1520 patients was 35.7 years of age. Most participants were male (77.7%) and Hispanic (98.8%). Forty-five percent (45.1%) of the population were single and only 21.9% were married; nevertheless, fifty one percent (51.7%) indicated to have children. 70% reported to be unemployed. Injecting Drug Usage appears as the first exposure mode (54.3%), followed by heterosexual contact cases (25.71%) and by men having sex with men (12.9%). The study of other risk practices revealed a large proportion of patients smoking tobacco (65.6%) and using alcohol (49.5%). Based on the 1993 CDC definition, forty-seven percent (47%) of the subjects had a clinical or immunological criterion to be considered as an AIDS case at first presentation. Among all AIDS cases, 440 patients presented with clinical AIDS (61.7%%) and 274 persons were classified as AIDS due to low CD4 counts alone (38.3%%). The most common AIDS defining conditions were: Pneumocystis carinii pneumonia (n = 201, 28.1%), Candidiasis Esophageal (n = 123, 17.2%), Toxoplasmosis (n = 95, 13.3%), Wasting syndrome (n = 68, 9.5%), and Tuberculosis (n = 68, 10.3%). CONCLUSIONS: The socio demographic and risk profile of AIDS patients in the present study is representative of the Puerto Rican AIDS population with regards to gender, age distribution, and risk scenario groups. This study revealed that a wide spectrum of social and behavioral vulnerabilities are impacting this population. A large proportion of patients is arriving to the health care facilities at a late stage of disease. Further studies including data from follow-up interviews will help to assess changes in the expression and evolution of the disease. PMID- 9419929 TI - Profile of a population using a primary health care center in Puerto Rico. AB - PURPOSE: To describe the services delivered by the Family Medicine Physicians at a Community Health Center. METHODS: All information from patient visits during the natural year 1996 were registered using a commercialized computer program. The information was gathered by different means: initial interview, physician's report, records, and personal interviews. RESULTS: A total of 13,203 visits were registered; this represent a total of 4,493 patients. Most of the patients were women, and with a mean age of 38. As expected, most of the patients have Medicaid. The most common conditions seen were hypertension, diabetes, and respiratory diseases. The mean number of visits during the year for almost all conditions was three. Most of the children and adolescents visit the Center due to respiratory conditions, while adults come due to hypertension, diabetes, and musculoskeletal conditions. PMID- 9419930 TI - Recent advances in cancer therapy. AB - The treatment of cancer has developed substantially from its conception in the first years of the 20th century. Since the introduction of alkylating agents during second World War, the oncology specialty has markedly grown. In the recent years, new drugs have been approved for the treatment of cancer. Such examples include the taxanes (Docetaxel and Paclitaxel), Vinorelbine, Irinotecan, Topotecan, Gemcitabine and Gliadel. We will discuss these new chemotherapuetic agents, their pharmacology, indications, toxicity and appropriate dosing. There is no doubt that further clinical research is needed to determine the optimal use of these agents. PMID- 9419931 TI - The investigation of emerging and re-emerging viral diseases: a paradign. AB - Emerging virus infections are defined as previously nonthreatening viruses that can decimate new populations by finding fresh hosts and vectors--often with the help of humans who introduce new species into virgen environment, Several etiologic agents of these diseases, some of the interacting factors that contribute to their development and the role of molecular medicine in their understanding is discussed. PMID- 9419932 TI - Jarcho-Levin syndrome: a new case report with unusual unexplained aortic root dilatation. AB - Since Jarcho and Levin described a condition involving extensive vertebral malformations and early death in 1938, many cases have been reported using multiple synonyms. Later, Solomon (3) proposed a subtype classification system to improve counseling concerning risk of recurrence, management, and prognosis. This is a report of a new Hispanic case with findings of spondylothoracic dysostosis and unusual aortic root dilatation. PMID- 9419933 TI - Which "Q" represent infarction? PMID- 9419934 TI - HIV/AIDS risk factors among adolescent students in Puerto Rico, 1994. AB - PURPOSE: Identify several HIV risk behaviors among adolescent students. METHODS: The sample (n = 3,648) was selected using a two-staged stratified cluster sampling design, and weighted to represent all junior high and high school students. RESULTS: About 28.8% of the students reported ever having sexual activity. Less than half of the sexually active (44.5%) used condoms during their last sexual activity; 27.6% used them always. Only 54.7% knew correctly > 75% of the HIV knowledge questions. A HIV risk scale was constructed using five risk factors. About 15.9% of the students did not have any risk factor, 36.2% had one, 47.9% had two or more. Males and high school students had significantly more risk factors. Half of the students will abstain from having sex next year because they don't want to get HIV/AIDS. CONCLUSIONS: It is important to implement effective HIV prevention programs for adolescents in order to change their attitudes and behaviors. PMID- 9419935 TI - Alcohol and/or cocaine effect on driving. PMID- 9419936 TI - Between two paradigms. PMID- 9419937 TI - Paradigm busters. PMID- 9419938 TI - Management of dental emergencies. PMID- 9419939 TI - Do we still care about children's teeth? PMID- 9419940 TI - Non-consultant career grade oral & maxillofacial surgeons. PMID- 9419941 TI - Asking the academics. AB - Assumptions are no match for real consultation. As part of the MRI, the BDJ conducted a special survey to see if academic dentists could predict the primary care research priorities of general dental practitioners. How would the predictions differ from the priorities of the GDPs themselves? PMID- 9419942 TI - Smoking cessation interventions for dental patients--attitudes and reported practices of dentists in the Oxford region. AB - OBJECTIVE: To investigate various aspects of dentists' beliefs and practices with respect to helping their patients stop smoking. DESIGN: Postal questionnaire survey conducted in 1996. SETTING: The general dental practitioners on the health authority lists of Berkshire, Buckinghamshire, Northamptonshire and Oxfordshire. SUBJECTS: The 869 dentists registered on 1 April 1996. RESULTS: A high response rate (78%; 674/869) was obtained. The majority of respondents (82%; 95% CI: 79, 85) thought dentists should encourage their patients to stop smoking although only 37% (95% CI: 34, 41) believed dentists to be effective in smoking cessation and even fewer (18%; 95% CI: 15, 21) routinely recorded their patients' smoking status. Of respondents, 51% (95% CI: 46, 55) said they always discussed smoking with patients who had periodontal problems but only 9% (95% CI: 7, 12) always did so with patients who had no major oral health problem. Newer graduates were more likely to routinely record their patients' smoking status (P = 0.02), and to think that doctors' advice (P = 0.001) and nicotine replacement therapy (P < 0.001) were effective in promoting smoking cessation. Dentists in mainly private practices were more active than those in NHS or mixed practices in recording patients' smoking status (P < 0.001) and in discussing smoking (P = 0.002). CONCLUSIONS: Most respondents thought that dentists should encourage their patients to stop smoking but few are active in this area. PMID- 9419943 TI - Factors influencing patient loyalty to dentist and dental practice. AB - OBJECTIVE: To establish the factors considered by adult regular dental attenders to be the most important in choosing to stay with a particular dentist or practice. DESIGN: An anonymous questionnaire to be completed by patients. SETTING: General dental practices in England and Wales with a significant proportion of patients seen under some form of private contract. SUBJECTS AND METHODS: 13 practices in England and Wales were selected to represent a regional cross-section of the country. Up to 100 successive patients visiting each practice during the summer of 1995 were invited to complete the questionnaire. 1003 questionnaires were returned. RESULTS: Notwithstanding regional, gender and socio-economic status related differences the most important factors were 'care and attention' rated as very important by 90% of respondents, 'pain control, 'dentist puts you at ease ', 'safety conscious' all rated as very important by 79 82% of respondents, and 'explanation of treatments' rated as very important by 73% of respondents. The importance attached to the dentist being 'safety conscious' suggests a change in emphasis from earlier studies. CONCLUSIONS: The factors rated most important reflect the dentist's behaviour and personal skills in devoting time and attention to the patient. PMID- 9419944 TI - The professional perception of orthodontic treatment complexity. AB - OBJECTIVE: To assess the current professional concept of orthodontic treatment complexity. DESIGN: Cross sectional survey of practitioners' views towards recently completed orthodontic treatments. SETTING: Specialist and non-specialist practitioners in General Dental Services in North Western and Mersey regions. The sampling was carried out between 1993-95. SUBJECTS AND METHODS: All practitioners undertaking orthodontic treatments in the GDS in the North Western and Mersey regions were invited to participate. Practitioners submitted consecutively started cases for scrutiny by analysis of study models and pre-treatment and post treatment questionnaires. 280 cases were collected. Data were analysed using multiple linear regression. MAIN OUTCOME MEASURES: The occlusal changes were assessed using the Index of Orthodontic Treatment Need and the Peer Assessment Rating. Self administered questionnaires employed 5-point Likert type scales to record practitioners' and patients' opinions. RESULTS: It appears that the concept of complexity is related to the occlusal changes which occur during orthodontic treatment. Attempts to define treatment complexity identifies factors which are of questionable validity and could be easily manipulated in practice setting. CONCLUSIONS: The present concepts of treatment complexity appear to be somewhat flawed. There is no occlusal index of complexity which is particularly satisfactory but as a crude interim measure 3 grades of complexity are proposed based on the PAR index. PMID- 9419945 TI - How to choose safe dental alloys. PMID- 9419946 TI - In at the deep end--an insight into scuba diving and related dental problems for the GDP. AB - Scuba diving is one of the fastest growing sports in the world. It is inevitable that the general dental practitioner (GDP) will have patients who participate in this sport and they should be aware of a number of problems that a diver can experience that are associated with the teeth and related structures. The aim of this article is to introduce the GDP to the dental problems associated with diving and make recommendations on dental care for scuba divers. PMID- 9419947 TI - Linkage analysis of families with autosomal dominant polycystic kidney disease by KG8-CA marker. AB - BACKGROUND: Autosomal dominant polycystic kidney disease (ADPKD) is one of the most common genetic diseases of human. Traditionally, ADPKD is diagnosed by ultrasonography, computed tomography (CT) or magnetic resonance imaging (MRI) of kidneys for the presence of renal cysts. Individuals who carry the defective gene but have not yet developed cysts in kidney may not be diagnosed. Genetic analysis reveals it to be caused mostly by a single-gene disorder of a genetic locus, designated PKD1. Recently, the genetic locus involving PKD1 has been identified on chromosome 16p13.3, and has been cloned and completely sequenced. METHODS: A pair of primers, KG8-CA, located between D16S84 and D16S125, was selected and synthesized for the polymerase chain reaction (PCR) to identify individuals who may carry the defective locus. The sequence of KG8-CA primers, was 5' CTCCCAGGGTGGAGGAAGGTG-3' and 5'-GCAGGCACAGCCAGCTCCGAG-3'. PCR products were analyzed in denaturing condition, using gel containing 8% acrylamide and 7M urea. Autoradiography was carried out to interpret the results. RESULTS: Four Chinese families with history of ADPKD showed different DNA patterns in individuals with ADPKD and in normal individuals. Among the members in four families with history of ADPKD, every individual shared a common DNA band, suggesting that this band was derived from normal PKD1 allele. On the other hand, individuals diagnosed to have ADPKD showed one or two additional DNA bands which migrated differently from the common DNA band and should therefore be derived from defective ADPKD allele. Previous studies have shown that the ADPKD allele is highly polymorphic, as was evident in these family studies. CONCLUSIONS: Among the members from these four families, some were clinically normal and had DNA pattern that was typical to patients with ADPKD. These individuals might carry the defective PKD1 allele but have not yet developed the ADPKD symptoms. Therefore, the method described in this study has diagnostic values for pre-symptomatic individuals as well as for patients already diagnosed with ADPKD. PMID- 9419948 TI - Core decompression in treating ischemic necrosis of the femoral head. AB - BACKGROUND: The application of core decompression in the treatment of ischemic necrosis of femoral head (INFH) is to preserve the joint at early stage of the disease. Although excellent results have been obtained in many series, its effectiveness is questionable because of high rates of failure. This study is to evaluate the effectiveness of this technique in our patients with INFH receiving core decompression as the sole treatment. METHODS: Although 91 patients were treated with core decompression in Veterans General Hospital-Taipei from 1980 to 1993, only 71 were considered as the valid cases after having been followed for 24 to 165 months (mean 56.7 months). Of 84 hips treated, 49 hips were steroid induced, 20 idiopathic, 14 alcohol related, and one related to trauma. Twenty five hips were at Steinberg stage I, 32 at stage II, nine at stage III, 17 at stage IV, and one at stage VI. RESULTS: The results were poor in 22 (26.19%) hips since the patients required hip prosthesis. Fair results were rated in 37 (44.05%) hips since progressive collapse occurred in the femoral head and the patients suffered from moderate hip pain. Good results were found in nine (10.71%) hips since the lesion showed no progression and the patients had only occasional pain. Excellent results were obtained in 16 (19.05%) hips since the patients had no pain and the lesion had no progressive change. The satisfactory rate in the nonsteroid-related hips (33% or 12/35) was significantly higher than in the steroid-related hips (27% or 13/49). The overall satisfactory result was only 30%. CONCLUSIONS: Although core decompression may not be effective in preventing progression of INFH, it may relieve temporary pain and delays the need for hip arthroplasty in some patients. PMID- 9419949 TI - Genetic polymorphism of monoamine oxidase B and susceptibility of Parkinson's disease. AB - BACKGROUND: Monoamine oxidase B (MAO-B) may play a role in the progression and cause of Parkinson's disease (PD), given consideration of the biochemistry and pathophysiology of the disease, and experiments on primates and humans. Assuming that the structural gene determines enzyme activity, an association study was undertaken to examine MAO-B genetic polymorphisms and look for the unique MAO-B gene alleles which occurred at a higher frequency in PD patients. METHODS: Sixty five PD patients, diagnosed according to the criteria set by a Core Assessment Program for Intracerebral Transplantations Committee, and 108 healthy controls were enrolled in this study. Genomic DNA was extracted from peripheral leukocytes by using a puregene kit. Polymerase chain reaction (PCR) was used to amplify the MAO-B genome. The PCR products were screened by restriction enzyme Hae III digestion and analyzed by single-stranded conformational polymorphism (SSCP). RESULTS: Two bands with different mobility shifts, defined as MAO-B allele 1 and allele 2, were observed in SSCP analysis. Neither genotypes nor allelic frequencies of MAO-B showed a significant difference between PD patients and controls in our study. CONCLUSIONS: Polymorphism of MAO-B genome was demonstrated in this study. It failed to show an association of a genetic marker with PD. However, this did not necessarily exclude the MAO-B locus from playing a role in causing PD because a polymorphism different from the one evaluated here may show some disease association. PMID- 9419951 TI - Complications of membrane-filtration plasma exchange. AB - BACKGROUND: Plasma exchange (PE) has been extensively used to treat a variety of disease states in the past three decades. Although PE is commonly thought to be a relatively safe procedure, a number of unpleasant side effects may occur. To highlight the need for continuing quality assurance in providing PE service, a retrospective review of PE records over a 13-year period was undertaken. METHODS: From April 1983 to March 1996, 694 therapeutic PE procedures, membrane-filtration type, were performed on a total of 157 patients in this hospital. Plasmaflo op 05(L) (Asahi Japan) was used as the plasma separator and fresh frozen plasma of one plasma volume as replacement fluid. The PE sheets for all procedures were reviewed as were medical charts in order to evaluate clinical efficacy. RESULTS: Totally 694 PEs in 157 patients, 84 male and 73 females, ranging in age from 1 to 75 years (median, 35 years), were treated. Patients received a median of 4 treatments (range, 1 to 34). The most frequent indications were severe jaundice (41.4%), systemic lupus erythematosus (23.6%), and myasthenia gravis (15.3%). The overall effective rate was 40.8%, and was best in neurological diseases (91.4% effective). Complications of PE were noted during 36% of the procedures involving 59.2% of the patients treated. The most common adverse reactions were paresthesia (12.7%), chills (10.2%), urticaria (8.5%), chest pain (5.9%), nausea (2.9%) and dyspnea (2.0%). Less common complications included: catheter infection (0.3%), catheter oozing (1.2%), dizziness (1.3%), headache (0.4%), muscle cramps (1.3%), vomiting (0.4%), abdominal pain (0.9%), fever (0.3%), hypotension (1.3%), bleeding (0.4%), consciousness change (0.7%) and respiratory arrest (0.4%). These complications were classified as "mild" (23%), "moderate" (11.4%) and "severe" (1.6%). No deaths occurred within 24 hours after PE. CONCLUSIONS: Although the use of membrane-filtration PE represents a valuable and relatively safe therapy, some life-threatening reactions do occur. PMID- 9419950 TI - The effects of ursodeoxycholic acid in patients with severe obstructive jaundice after drainage procedure. AB - BACKGROUND: The aim of this study is to evaluate the therapeutic effect of ursodeoxycholic acid (UDCA) for patients with severe obstructive jaundice after drainage procedure. METHODS: From September 1993 to December 1994, patients admitted with severe obstructive jaundice (serum bilirubin > 15 mg/dl) and successful drainage were enrolled into our study. They were randomly divided into two groups to receive UDCA 600 mg per day (UDCA group) or a placebo (placebo group) until operation or discharge. Bile drainage amount, clinical symptoms and signs and adverse effects of drugs were recorded daily. Biochemical tests, including albumin, total bilirubin, cholesterol, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, gamma-glutamyl transpeptidase and prothrombin time, were checked first, on the third and seventh days, and then two weeks after drainage. RESULTS: Thirty-eight patients (M/F = 36/ 2, mean age 66.6 +/- 6.4 years) completed the study, 18 in the UDCA group and 20 in the placebo group. There were no differences in age, sex, causes of obstructive jaundice or methods of drainage procedure between the UDCA and placebo groups. Improvement in biochemical test results were noted in both groups. However, bile drainage amount and changes in liver biochemical test, especially the decrement of serum bilirubin, were not significantly different between both groups. CONCLUSIONS: UDCA seemed not to benefit patients with severe obstructive jaundice after successful drainage. PMID- 9419952 TI - Role of foramen ovale electrodes in presurgical evaluation of intractable complex partial seizures. AB - BACKGROUND: The value of intracranial electrodes such as depth electrodes and subdural grids for intracranial electroencephalographic (EEG) recording in patients with intractable epilepsies has been well recognized. A new technique, foramen ovale electrode (FOE) implantation, was first introduced by Wieser in 1984 for the lateralization of bilateral mesiotemporal lobe (MTL) onset of seizures. METHODS: Since October 1993, a multipolar, three-contact FOE has been used in 12 intractable epileptic patients for presurgical evaluation. The reasons for FOE implantation included bilateral MTL onset of seizures recorded by extracranial EEGs in nine patients, and extracranial EEG abnormalities inconsistent with the results of magnetic resonance imaging (MRI), positron emission tomogram (PET) or Wada test in three patients. Under general anesthesia, the FOEs were implanted according to the technique introduced by Kirschner, using Barters landmarks. RESULTS: After long-term telemetry recording with FOE, seven patients revealed clear onset of seizures originating from one side of the MTL and underwent anterior temporal lobectomy (ATL). Two patients had seizures of bilateral MTL onset. However, they received ATL due to predominantly unilateral interictal epileptiform discharges (EDs) and/or MRI and PET abnormalities. Seven (78%) of the nine operated patients became seizure-free after ATL. Three patients were considered not operable because two had multifocal onset of seizures and one had seizures with independent bilateral MTL onset. No serious complication resulted from implantation of FOE in this series. CONCLUSIONS: The semi-invasive technique of FOE is reliable for lateralization of bilateral MTL onset of seizures which are often not clearly recorded by extracranial EEGs. This procedure is safe and can be an alternative to invasive implantation of depth electrodes and subdural grids. PMID- 9419954 TI - MRI-guided stereotatic aspiration for the treatment of deep-seated tentorial empyema: a case report. AB - Tentorial empyema is a rare intracranial infection. In the past, most authors treated the subdural empyema by craniotomy, craniectomy or burr hole drainage. In the case presented, magnetic resonance imaging (MRI)-guided stereotatic aspiration was used to treat deep-seated extensive tentorial empyema successfully, via a simple burr hole to avoid any brain resection and retraction. This operation method is relative simple and effective for treating deep-seated intracranial empyema, significantly reduces the risk of injury to the lateral temporal lobe cortex or transection of optic radiations. On the other hand, it must be emphasized that contrast-enhanced MRI scan may offer a more sensitive and better diagnostic procedure than contrast enhanced computed tomography (CT) scan. PMID- 9419955 TI - Tuberculous arthritis of sacroiliac joint with abscess formation: a case report. AB - Extrapulmonary tuberculosis at times appears to be an unusual presentation of the disease. Its incidence has not decreased, though a slow decline in pulmonary tuberculosis has been observed over the past decades around the world. The diagnosis of extrapulmonary tuberculosis is still rather difficult today. Tuberculosis of the bones and joints, which constitutes a smaller part of extrapulmonary tuberculosis, is relatively rare in documented cases and can yield variable clinical manifestations. This study describes a young male patient who developed a rare condition of tuberculous arthritis of the right sacroiliac joint, with an abscess formation over the anterior aspect of the right iliac fossa and iliopsoas muscle. The diagnosis was made by roentgenographic examinations and proved by microscopic confirmation of the presence tuberculous bacilli in drainage from the involved foci. Antituberculosis therapy was begun thereafter. Symptoms improved and the patient was regularly followed up at the Outpatient Department. It is suggested that in patients with prolonged fever and possible Mycobacterium tuberculosis infections, repeated roentgenographic and microbiological examinations are mandatory. PMID- 9419956 TI - Major depressive disorder with delusion of having AIDS: a case report. AB - Acquired immune deficiency syndrome (AIDS) has become a topic of increasing public concern. Several conditions have been described in patients who are afraid that they have acquired AIDS, including depression, somatization disorders, hypochondriasis, adjustment disorders and various psychoses. This paper presents a case study of a young man with persistent fear of having AIDS. In spite of negative HIV test results, the patient still insisted that he suffered from AIDS. The psychiatric diagnosis was major depressive disorder with delusion of having AIDS. The patient was treated with fluoxetine 40 mg qd and sulpiride 400 mg qn. After two weeks on medication, his fear of AIDS subsided and he improved remarkably. The most important intervention in patients with delusion of having AIDS is to identify and treat the underlying psychopathology. The following case is representative of our experience. PMID- 9419953 TI - Pyogenic myositis: caused by viridans streptococci in an adult with tetralogy of Fallot. AB - A 34-year-old man came to this Emergency Room because of fever, swelling and pain in the right thigh. Tetralogy of Fallot with bicuspid pulmonary valve was diagnosed after serial examination, and computed tomography of the right thigh revealed pyogenic myositis. Surgical drainage of the right thigh abscess and a further pus culture yielded viridans streptococci. There has been no record in the medical literature of a pyogenic myositis caused by viridans streptococci in an adult tetralogy of Fallot. This is thus the first case reported. PMID- 9419957 TI - Leprosy in England and Wales. PMID- 9419958 TI - Coenzyme Q10: a vital therapeutic nutrient for the heart with special application in congestive heart failure. AB - Vitamin coenzyme Q10 is a critical adjuvant complementary therapy for patients with congestive heart failure, especially when traditional medical therapy is unsuccessful. The following case studies, with systolic and/or diastolic dysfunction, demonstrate the effectiveness of coenzyme Q10 in improving quality of life, as well as survival. PMID- 9419959 TI - Puerto Rican patients travel to Puerto Rico: assessing the effect on clinical care. AB - OBJECTIVE: To examine travel habits of Puerto Rican patients and assess the potential effect of this travel on their health care. DESIGN: Interview and survey of patients. SETTING: Urban medical clinic. PATIENTS: Two hundred consecutive, self-identified Puerto Rican patients presenting for follow-up care. INTERVENTION: Immediately prior to a follow-up office visit, patients were interviewed in either Spanish or English. MEASUREMENTS: The patients' age, sex, education level, employment status, and place of birth were recorded. The patients were asked questions concerning the principle place of residence of their family members, their ability to speak English, and their preferences in television and radio programs. Patients who had visited Puerto Rico were asked about the duration and purpose of their most recent trip and about the health care they received in Puerto Rico. Chi-square testing was applied to categorical data and t tests for continuous data; P values were calculated using the SPSS statistical analysis program. RESULTS: Of the 200 subjects, 110 (55%) had traveled to Puerto Rico in the last five years and 90 (45%) had not. The patients who traveled were more likely to have been born in Puerto Rico, less likely to speak English, and less likely to listen to English-language programs. A majority of the patients who traveled to Puerto Rico visited for a month or less (80%) and did not experience a change in health care (78%). In comparison, however, a majority (59%) of the patients who visited for longer than a month did experience a major change in their health care status (P < .00001). CONCLUSIONS: Puerto Rican patients, particularly those born in Puerto Rico with stronger cultural "ties" to the island, frequently return to Puerto Rico. Patients who visit for longer than a month often experience changes in care which are likely to have significant effect on their health. Clinicians caring for Puerto Rican patients should ask about upcoming visits to Puerto Rico and take steps to assure continuous and coordinated medical care. PMID- 9419960 TI - Serotonin syndrome: case report and review of the literature. AB - We present the case of a 35-year-old woman who developed serotonin syndrome after receiving a single dose of the cyclic antidepressant imipramine (Tofranil). She was already being treated for depression with paroxetine (Paxil), a selective serotonin reuptake inhibitor. Two hours after receiving imipramine, the patient developed tachycardia, delirium, bizarre movements, and myoclonus, all classic findings of serotonin syndrome. Her antidepressants were discontinued and she was treated with intravenous fluids, sedation, and a short course of cyproheptadine, a serotonin receptor antagonist. All symptoms resolved completely within 24 hours. In this case report, we review the drug interactions that can precipitate serotonin syndrome, and give recommendations for the diagnosis and treatment of this potentially fatal disorder. PMID- 9419961 TI - Peripheral parenteral nutrition: an underused nutritional modality. PMID- 9419962 TI - Ethical standards--Part Two. PMID- 9419963 TI - In response to Dr. Chotkowski's letter. PMID- 9419964 TI - Ethics in managed care. PMID- 9419965 TI - Academia's chilly climate for primary care. PMID- 9419966 TI - Proliferation of human malignant astrocytomas is dependent on Ras activation. AB - Overexpression and activation of receptor tyrosine kinases, such as platelet derived growth factor receptors (PDGFRs) and epidermal growth factor receptor (EGFR), leads to proliferation of human malignant astrocytoma cells. Although oncogenic mutations affecting Ras are not prevalent in human malignant astrocytomas, we have investigated whether levels of activated Ras.GTP might be elevated in these tumors secondary to the mitogenic signals originating from activated receptor tyrosine kinases. In support of this hypothesis high levels of Ras.GTP, similar to those found in oncogenic Ras transformed fibroblasts, were present in four established human malignant astrocytoma cell lines which express PDGFRs and EGFR, and 20 operative malignant astrocytoma specimens. Stimulation of PDGFR's and EGFR's induced tyrosine phosphorylation of the Shc adaptor protein and its association with Grb2, suggesting a mechanism by which Ras may be activated in human malignant astrocytoma cells. Furthermore, blocking Ras activation by expression of the Ha-Ras-Asn17 dominant-negative mutant, or by farnesyl transferase inhibitors, decreased in vitro proliferation of the human astrocytoma cell lines. These results support the hypothesis that proliferative signals from receptor tyrosine kinases expressed by human malignant astrocytoma cells utilize the Ras mitogenic pathway. Pharmacological inhibitors of the Ras pathway may therefore be of therapeutic value in these presently terminal tumors. PMID- 9419967 TI - Bcl-2 inhibits p53 nuclear import following DNA damage. AB - Bcl-2 is an integral membrane oncoprotein that localizes to membranes of the mitochondria, endoplasmic reticulum, and nuclear envelope. Bcl-2 is a member of a family of cell death regulators and functions to inhibit apoptosis. Using confocal microscopy and immunoblotting we show that the ability of bcl-2 to suppress cell death following genotoxic damage can be a consequence of inhibiting nuclear import of induced wild-type p53 protein. Our data suggests that the ability of bcl-2 to modulate trafficking events is not cell type specific. These data support a 'gatekeeper' mechanism for cell death suppression by bcl-2. PMID- 9419968 TI - Human chromosome 7 carries a putative tumor suppressor gene(s) involved in choriocarcinoma. AB - Choriocarcinoma developed from a complete hydatidiform mole has an unique genetic feature that involves monoallelic contribution from the paternal genome. To determine the chromosome carrying putative tumor suppressor gene(s), microcell hybrids were isolated following fusion of choriocarcinoma cells with microcells from mouse A9 cells containing a single human chromosome (1, 2, 6, 7, 9 or 11). Microcell-hybrids with the introduction of chromosome 7 were suppressed or modulated for tumorigenicity and exhibited altered in vitro growth properties. Introduction of chromosomes 1, 2, 6, 9 or 11 had no effect. Tumorigenic revertants isolated from microcell-hybrids with the introduced chromosome 7 contains reduced numbers of chromosome 7. These findings suggest that chromosome 7 contains a putative tumor suppressor gene(s) for choriocarcinoma. Alterations in tumorigenic phenotypes seen in microcell-hybrids were not associated with the presence of either ERV3 or H-plk locus located on the introduced chromosome 7, indicating the putative tumor suppressor gene(s) is outside of ERV3 and H-plk gene loci. Furthermore, we obtained evidence to define a critical region on chromosome 7 (7p12-7q11.23) that was frequently lost in surgically removed choriocarcinoma tissues and cell lines. Using a panel of microsatellite markers, biallelic deletions were observed, which strongly suggests the presence of a tumor suppressor gene(s) within this critical region. PMID- 9419969 TI - Oncogenesis and altered differentiation induced by activated Ras in neuroblasts of transgenic mice. AB - Sympathetic neurons, enteric neurons and adrenal chromaffin cells all derive from the neural crest. During development these cells migrate, proliferate, survive and differentiate in a highly controlled fashion influenced by local signals encountered during their migration. Aberrations of these processes are responsible for a variety of developmental defects and malignancies. Many of the environmental signals influencing these precursor cells activate receptor tyrosine kinases that can signal, at least in part, via Ras pathways. To assess the extent to which Ras can alter neuroblast cell number and fate in vivo, we expressed activated H-Ras in transgenic mice using the dopamine-beta-hydroxylase promoter, which directs expression to these cells prior to and after their differentiation. Ganglioneuromas and occasional neuroblastomas formed in the adrenal gland and preaortic sympathetic ganglia. Curiously, neurons of the superior cervical ganglia and the gut were largely unaffected despite demonstrated expression of activated Ras. The sensitivity of preaortic sympathetic neurons and adrenal chromaffin cells to the effects of oncogenes such as Ras may explain the predilection of neuroblastomas in humans to these sites. The ability to analyse neuroblastoma development in these mice may shed light on the molecular basis of certain types of human neuroblastoma. PMID- 9419970 TI - Repression of hepatitis B virus (HBV) transgene and HBV-induced liver injury by low protein diet. AB - Persistent infection with hepatitis B virus (HBV) is one of the primary risk factors for human hepatocellular carcinoma (HCC). In a human ecological study, we have shown that, in addition to HBV, animal food consumption also significantly contributes to the variance of HCC. To test the interacting effect of HBV and animal food consumption on the development of HCC, we investigated HBV expression in HBV transgenic mice fed three levels of casein diet. HBV expression in transgenic animals was substantially inhibited when dietary casein was reduced from the traditional level of 22% to the level of 6%. Northern analysis revealed that suppression of HBV was derived from both the upstream albumin promoter and the internal HBV promoter. Immunochemical staining of liver sections indicated that only a few hepatocytes around the central vein expressed viral surface antigen (HBsAg) in the 6% casein animals, whereas virtually all hepatocytes stained positively for HBsAg in the 22% dietary casein animals. Serum HBsAg concentrations at 4 months were increased by 1.6-, 2.1-, and 5.1- fold over baseline for animals fed the 6%, 14%, and 22% casein diets, respectively. Correspondingly, liver injury was much less severe in animals fed 6% casein diet than in those fed 14% and 22% casein diets. These results demonstrate that a low casein diet is a potent suppresser of HBV transgene and HBV-induced liver injury, suggesting that diet management may be a practical means to aid in the control HBV infection. PMID- 9419971 TI - Differential expression of NDF/neuregulin receptors ErbB-3 and ErbB-4 and involvement in inhibition of neuronal differentiation. AB - Two receptor tyrosine kinases, ErB-3 and ErbB-4, mediate signaling by Neu differentiation factors (NDFs, also called neuregulins), while ErbB-1 and ErbB-2 serve as co-receptors. We show that the two NDF/neuregulin receptors differ in spatial and temporal expression patterns: The kinase-defective receptor, ErbB-3, is expressed primarily in epithelial layers of various organs, in the peripheral nervous system, and in adult brain, whereas ErbB-4 is restricted to the developing central nervous system and to the embryonic heart. An example of alternating expression of the two receptors is provided by the developing cerebellum: During postnatal cerebellar development, ErbB-4 expression slightly decreases along with a decline in NDF transcription, whereas ErbB-3 expression commences after the peak of neurogenesis. To study functional differences, we established primary brain cultures and found that ErbB-3 was expressed only in oligodendrocytes, whereas ErbB-4 expression was shared by oligodendrocytes, astrocytes and neurons. Blocking the action of endogenous NDF in vitro, by using a soluble form of ErbB-4, accelerated neurite outgrowth in both primary cultures and in neuronal-type cultures of the P19 teratocarcinoma, suggesting an inhibitory effect of NDF on neural differentiation. Apparently, ErbB-3 is associated with proliferation of P19 cells, whereas ErbB-4 correlates with a differentiated phenotype. We conclude that the two NDF receptors play distinct, rather than redundant, developmental and physiological roles. PMID- 9419972 TI - Resistance of MCF7 human breast carcinoma cells to TNF-induced cell death is associated with loss of p53 function. AB - We have investigated the relationship between the development of tumor resistance towards the cytotoxic action of tumor necrosis factor-alpha (TNF) and p53 function, using the TNF-sensitive MCF7 human breast adenocarcinoma cell line and two TNF-resistant sublines, MCF7/R-A1 and MCF7/Adr. Use of single-strand conformation polymorphism (SSCP) analysis and DNA sequencing shows that MCF7 has a wild-type p53 gene, whereas both TNF-resistant sublines exhibit mutant p53. This includes a point mutation R280K in MCF7/R-A1 cells, and a point mutation at the splicing acceptor site on the upstream border of exon 5 resulting in a 21 pb deletion in MCF7/Adr cells. These mutations result in loss of p53 capacity to transactivate FASAY (functional assay in yeast). In contrast to what is observed for parental MCF7 cells, treatment of resistant sublines with TNF or gamma irradiation fails neither to induce the expression of the p53-regulated gene products p21waf1/CIP1 and MDM2, nor to arrest the cells in the G1 phase of the cell cycle. Disruption of p53 wild-type function in MCF7 cells by transfection with human papillomavirus type-16 E6 gene, leads to abrogation of the cytotoxic, but not the cytostatic activity of TNF. Altogether, our results strongly suggest that wild-type p53 is involved in cytotoxic action of TNF, and point out that loss of p53 function contributes to resistance of tumor cell to TNF-induced killing. PMID- 9419973 TI - Distinct roles for DH and PH domains in the Lbc oncogene. AB - Members of the Dbl-homology (DH) family of proteins promote guanine nucleotide exchange on Rho GTPases. Lbc, which specifically acts on Rho but not Rac or Cdc42, was isolated as a transforming oncogene and is composed of a DH and a Pleckstrin-homology (PH) domain. We show here that the Lbc DH domain alone is capable of stimulating new DNA synthesis in quiescent fibroblasts and Rho dependent actin stress fiber assembly. However, the PH domain is required for subcellular localization of Lbc along actin stress fibers and for efficient transformation of NIH3T3 cells. The results show that, in contrast to other Dbl homology proteins, the PH domain of Lbc is dispensable for activation of Rho in vivo. The PH domain-dependent subcellular localization of Lbc may, however, be important for growth factor activation of endogenous Lbc and for oncogenic transformation. PMID- 9419974 TI - Induction of a beta-catenin-LEF-1 complex by wnt-1 and transforming mutants of beta-catenin. AB - Signal transduction by beta-catenin involves its posttranslational stabilization and import to the nucleus where it interacts with transcription factors. Recent implications for beta-catenin signaling in cancer prompted us to examine colon cancer cell lines for the expression of LEF-1, a transcription factor that binds to beta-catenin. The analysis of several cell lines revealed the expression of LEF1 mRNA and a constitutive association of the LEF-1 protein with beta-catenin. In contrast to the colon cells, PC12 and 293 cells did not contain a beta-catenin LEF-1 complex, even though both proteins were detected in cell lysates. In these cells, the association of endogenous LEF1 and beta-catenin was induced by stimulation with the wnt-1 proto-oncogene. The complex formed following transient stimulation with wnt-1 and also persisted in cells stably expressing wnt-1. Ectopic overexpression of beta-catenin in 293 cells also induced the assembly of the beta-catenin-LEF-1 complex and activated gene transcription from a LEF-1 dependent promotor. Expression of mutant oncogenic forms of beta-catenin identified in cancer cells resulted in higher levels of transcriptional activity. The results suggest that a cancer pathway driven by wnt-1, or mutant forms of beta-catenin, may involve the formation of a persistent transcriptionally active complex of beta-catenin and LEF1. PMID- 9419975 TI - Epiregulin binds to epidermal growth factor receptor and ErbB-4 and induces tyrosine phosphorylation of epidermal growth factor receptor, ErbB-2, ErbB-3 and ErbB-4. AB - Epiregulin is a member of the epidermal growth factor (EGF) family, and has certain characteristics that are different from that of EGF, including mitogenic responses and binding to EGF receptor (EGFR). Epiregulin may also have another cell surface receptor and/or induces different receptor heterodimerizations for intracellular signaling. We investigated the binding ability of epiregulin to four ErbB family receptors using four human breast carcinoma cell lines that expressed different subsets of receptors. Chemical cross-linking experiments showed that [125I]epiregulin directly bound to each of EGFR and ErbB-4 but not to ErbB-2 and ErbB-3. Furthermore, although epiregulin stimulated tyrosine phosphorylation of all four ErbB receptors, the main intracellular signal was mediated by ErbB-4 and/or EGFR. The pattern of activation of ErbB family receptors was different from that of other EGF-related ligands. Our findings indicate that ErbB-4 and EGFR are receptors for epiregulin, and suggest that EGF related ligands transduce signals for different biological responses by the hierarchical mechanism. PMID- 9419976 TI - Bcl-2 inhibits c-fos induction by calcium. AB - Transient elevation of cytosolic Ca2+ induces the expression of a variety of genes involved in cell growth and transformation, including the early response gene c-fos. Previously, we reported that Bcl-2 inhibits the transient elevation of cytosolic Ca2+ induced by thapsigargin (TG), a selective inhibitor of the endoplasmic reticulum-associated Ca2+-ATPase. Therefore, to determine if the effect of Bcl-2 on cytosolic Ca2+ elevation modulates Ca2+ signaling, we investigated the induction of c-fos by TG in WEHI7.2 mouse lymphoma cells, control transfectants (WEHI7.2-neo), and transfectants that stably express a high level of Bcl-2 (W.Hb12 and W.Hb15). TG induced 20-fold elevation of c-fos mRNA in WEHI7.2 and WEHI7.2-neo cells, but c-fos mRNA induction by TG was only fivefold in W.Hb12 and W.Hb15 cells. In contrast, phorbol 12-myristate acetate induced marked c-fos mRNA elevation in both WEHI7.2 and W.Hb12 cells, indicating that the inhibitory effect of Bcl-2 is selective for induction of c-fos by Ca2+. To measure c-fos promoter activity, WEHI7.2 and W.Hb12 cells were transiently transfected with a c-fos promoter-luciferase reporter plasmid. TG induced c-fos promoter activity in WEHI7.2 cells, but not in W.Hb12 cells. In WEHI7.2 cells, the signal for c-fos induction by TG was cytosolic Ca2+ elevation, as the increase in both c-fos mRNA level and promoter activity were prevented by lowering extracellular Ca2+ concentration, a condition that inhibits cytosolic Ca2+ elevation by reducing the TG-mobilizable Ca2+ pool. In summary, the findings indicate that Bcl-2 regulates Ca2+ signaling. PMID- 9419977 TI - Engineered mutants of pRB with improved growth suppression potential. AB - We have constructed a panel of substitution mutants which affect one or more of the putative cdk target sites of the RB protein. We have examined the activity of these mutants relative to wild-type RB by both a transcriptional repression assay and by measuring growth suppression in vitro. We find that some phosphorylation site mutants of pRB can repress E2 transcription more strongly than wild-type RB. These mutants are partially resistant to phosphorylation by cdks and can arrest tumor cells in G1 in vitro. Our results indicate a functional correlation between the ability to repress E2F-dependent transcription and the ability to suppress tumor cell growth in vitro. In addition, we describe two classes of RB mutants: N terminal truncated p56RB and a novel mutant of RB containing multiple substitutions near its nuclear localization signal. Both classes of RB mutants have greater activity than the wild-type protein. Because RB is a key regulator of cell cycle progression, expression of a more potent, phosphorylation resistant RB may have utility in both RB(-/-) and RB(+/+) tumors as well as in hyperproliferative disorders. PMID- 9419978 TI - Involvement of p21 in mitotic exit after paclitaxel treatment in MCF-7 breast adenocarcinoma cell line. AB - It has been shown recently that expression of p21 is enhanced by paclitaxel. This cytotoxic compound induces mitotic spindle damage resulting in blockade of the mitotic cell cycle associated or not with apoptotic cell death. In the present study, we showed that, in MCF-7 cells, paclitaxel induced accumulation of p21 in cells with a G2/M DNA content, corresponding to cells either in abnormal mitosis or in an interphase-like state (decondensed chromatin) with multiple nuclei. In MCF-7 cells, the increase in p21 was subsequent to the mitotic arrest and was associated with the exit from abnormal mitosis leading to formation of cells with micronuclei. In this cell line, we noted a relationship between the elevation of p21 expression and the inhibition of p34cdc2 activity. High levels of p21 protein were also found to be associated with inactive p34cdc2/cyclin B protein complex after treatment with paclitaxel. Treatment with p21 antisense oligonucleotide partially blocked induction of p21 expression by paclitaxel and significantly reduced survival of MCF-7 cells exposed to this agent. In NIH-OVCAR-3 cells, which are deficient in basal and paclitaxel-induced p21 expression, paclitaxel led to a prolonged activation of p34cdc2 and a delayed mitotic exit associated with apoptotic cell death. These observations suggest that p21 is not required for the mitotic arrest in response to paclitaxel, but argue in favor of a role for this inhibitor in facilitating the exit from abnormal mitosis. This effectively enhances cell survival after paclitaxel-induced spindle damage. PMID- 9419980 TI - Norgestomet implants prevent pregnancy in beef heifers on pasture. AB - The efficacy of erodible norgestomet implants for preventing pregnancy in postpubertal heifers was evaluated in two experiments at five locations each. Heifers (n = 896) within each study location were stratified by weight and allotted randomly to receive an ear implant containing either 0, 24, 36, or 48 mg of norgestomet (d 0). Heifers were exposed to fertile bulls immediately after implantation for 75 d (d 0 to 74) in Exp. 1 (n = 476) or for 80 d (d 75 to 154) in Exp. 2 (n = 420). Weights were recorded on d 0 and 74 (Exp. 1 and 2) and d 154 (Exp. 2). Each heifer was palpated rectally for pregnancy at the end of each experiment. Pregnancy rates were higher (P < .01) for control heifers (0 mg implant) than for heifers that received 24, 36, or 48 mg of norgestomet. In Exp. 1, pregnancy rates were 96, 29, 6, and 4% for heifers that received 0, 24, 36, and 48 mg implants of norgestomet, respectively. In Exp. 2, pregnancy rates were 85, 36, 19, and 9% for heifers that received 0, 24, 36, and 48 mg implants of norgestomet, respectively. Estrous activity during the first 3 wk of bull exposure was reduced (P < .05) among heifers that received norgestomet implants compared to control heifers but was not completely abolished at any dosage in Exp. 1. During the first 75 d of Exp. 1 and 2, heifers treated with 36 or 48 mg norgestomet implants gained weight faster (P < .05) than control heifers. Combined across both experiments, ADG during the first 74 d were .53, .56, .59, and .60 kg/d for heifers treated with 0, 24, 36, and 48 mg implants of norgestomet, respectively. These data indicate that norgestomet implants increased rate of weight gain, reduced estrous activity, and reduced the occurrence of pregnancy in heifers on pasture. PMID- 9419979 TI - Drastic genetic instability of tumors and normal tissues in Turcot syndrome. AB - Turcot syndrome is characterized by an association of malignant brain tumors and colon cancer developing in the patient's teens. Since the mechanism of carcinogenesis in Turcot syndrome is still unclear, we analysed genetic changes in tumors from a Turcot patient with no family history of the condition. All tumors, including one astrocytoma, three colon carcinomas, and two colon adenomas, exhibited severe replication error (RER), and all colon tumors showed somatic mutations at repeated regions of TGFbetaRII, E2F-4, hMSH3, and/or hMSH6 genes. Somatic APC mutations were detected in three of three colon carcinomas, and somatic p53 mutations were detected in the astrocytoma and two of three colon carcinomas, both of which showed two mutations without allele loss. We also found that normal colon mucosa, normal skin fibroblasts and normal brain tissue from this patient showed respective high frequencies of RER, in contrast to usual HNPCC patients in which RER was very rare in normal tissues. These results suggest that extreme DNA instability in normal tissues causes the early development of multiple cancer in Turcot syndrome. A missense mutation (GAG to AAG) at codon 705 of hPMS2 gene was detected in one allele of this patient, which was inherited from his mother without tumors. Additional unknown germline mutation may contribute to the genetic instability in normal tissues. PMID- 9419981 TI - Effects of programmed gain strategies on performance and carcass characteristics of steers. AB - In Trial 1, 161 Angus x Simmental crossbred steers (initial BW 305 +/- 1.0 kg) were used in a completely randomized design experiment to determine the effects of intake restriction and programmed gain on cattle performance and carcass composition and characteristics. Five feeding systems were tested using step-wise increases in programmed intake level. Initially steers were fed to gain 1.13 kg/d. Intake was then increased to achieve a gain of 1.36 kg/d. At the end of the feeding period, steers had ad libitum access to feed. Duration of intake restriction and the period of unrestricted intake was varied. Feeding steers at restricted intakes and then increasing daily gain by increasing feed intake using four different schedules all reduced (P < .05) daily feed intake and total feed intake compared with providing ad libitum access to feed throughout the trial. Furthermore, daily feed efficiency was increased (P < .05) by two of the feeding systems compared with offering ad libitum access to feed throughout the trial. The feeding system used did not affect (P > .10) quality grade of the carcasses. In Trial 2, 77 individually penned Angus x Simmental crossbred steers (initial BW 273 +/- 1.2 kg) were used to determine the effects of various feed intake restriction systems. For systems 1 through 4, multiple periods of restriction and realimentation were investigated; the duration and magnitude of restriction were varied. Feed intake was not restricted for steers in system 5. The feed restriction systems used in this experiment did not result in decreased total feed intake or changes in carcass composition as compared with offering ad libitum access to feed. Reducing total energy intake seems to be a prerequisite to altering feed efficiency of steers in limit-feeding systems. PMID- 9419982 TI - Benefit of multiple trait selection to increase reproductive traits: experimental evidence from golden hamsters. AB - Fifteen generations of selection were conducted to study responses for litter size at birth (LSB), weight at weaning of standardized litter (LWW), and individual body weight at 8 wk of age (BW8) using golden hamsters as an experimental model for pigs. The experiment involved three lines: selection on an aggregate breeding value of LSB, LWW, and BW8 (line W); selection on an aggregate breeding value of LSB and LWW (line R); and a randomly selected control (line C). Selection in W and R was based on breeding values from a multiple trait animal model. Restricted maximum likelihood with an animal model was used to estimate genetic parameters and genetic trends. Heritability estimates for LSB, LWW, and BW8 were .10, .47, and .52, respectively, and genetic correlations between traits were all positive. The mean estimated breeding value (EBV) for LSB in generation 15 was +2.2 pups in W and R. The mean EBV for LWW in generation 15 was +318 g for W and +174 g for R, and for BW8 means were +64 g and +24 g, respectively. Average inbreeding at generation 16 was 13.4, 19.5, and 8.0% for W, R, and C, respectively. Including BW8 in the selection criterion reduced inbreeding and had a beneficial effect on selection responses in LSB, LWW, and BW8. PMID- 9419983 TI - Maternal performance differences between porcine stress syndrome-normal and carrier Landrace females. AB - Differences between porcine stress syndrome (PSS) normal (NN) and carrier (Nn) Landrace dams were determined for adjusted number of pigs born alive, adjusted number of pigs at 21 d, adjusted 21-d litter weight, proportion of pigs surviving to 21 d, and farrowing interval. Data were analyzed from a total of 841 females, 623 normal (NN) and 218 carriers (Nn) having 2,231 and 869 records, respectively. Three susceptible (nn) females from two herds were dropped from the analysis because of their small contribution to the total number of records. Frequency of the recessive PSS allele ranged from .07 to .28 in the nine herds involved in this study. Data were adjusted using Landrace breed-specific adjustments and analyzed with mixed-model derivative-free REML procedures fitting the dams' PSS genotype as a fixed effect in the model. Only females having two or more successive parities were used in the analysis of farrowing interval, resulting in a reduction of total records analyzed to 2,201 (1,564 NN and 637 Nn records) from 632 females (445 NN and 187 Nn females). No differences between NN and Nn dams were observed for adjusted number of pigs born alive, adjusted number of pigs at 21 d, adjusted 21-d litter weight, proportion of pigs surviving to 21 d, and farrowing interval. The results of this investigation indicate no significant maternal performance differences between PSS NN or Nn Landrace dams. PMID- 9419984 TI - Nonlinear genetic relationships between traits and their implications on the estimation of genetic parameters. AB - To estimate nonlinear genetic relationships between traits, formulas based on the paternal halfsib analysis theory were derived. To illustrate the usefulness of the formulas, a series of data sets with halfsib structure for some preselected parameters and sample size situations were generated by means of Monte Carlo techniques. When a nonlinear relationship in the form of a polynomial relationship of degree two is present, the linear and quadratic regression coefficients can be estimated from a paternal halfsib analysis without bias. Some of the traditional linear genetic parameters need correction; however, their value is limited if the relationship between two traits is nonlinear. Although regression coefficients may be estimated appropriately in many situations, the application of the described method is restricted to situations in which the causal flow between the traits involved is clear and the heritability of the determining trait is larger than approximately .10. Further work should be directed to investigation of possibilities for including such parameters in selection decisions in a formalized way. PMID- 9419985 TI - Phenotypic and genetic parameters for longissimus muscle fiber characteristics in relation to growth, carcass, and meat quality traits in large white pigs. AB - A total of 383 barrows and gilts from a French Large White experimental herd were slaughtered at 100 kg BW. Samples of longissimus muscle were taken to categorize myofibers according to their contractile (I, IIA, and IIB) and metabolic (oxidative and nonoxidative) properties. Myofiber percentages, cross-sectional areas (CSA), and relative areas were measured. Growth rate, carcass composition, muscle chemical composition, metabolic enzyme activities, and meat quality traits were also measured to estimate phenotypic and genetic correlations between these traits and myofiber characteristics. Genetic parameters were estimated using a REML procedure applied to an individual animal model. Heritabilities of fiber traits were moderate to high (h2 = .20 to .59). Highest heritabilities were found for type I fiber percentage (h2 = .46 +/- .11), type IIBw fiber percentage (h2 = .58 +/- .11), and type I fiber cross-sectional area (h2 = .59 +/- .10). For a given fiber type, the relative area was phenotypically and genetically more closely related to the percentage than to the CSA. Phenotypic correlations between fiber type composition and other traits were low. Genetically, growth rate, carcass leanness, and loin eye area were positively related to fiber CSA. Intramuscular fat content was not related to fiber type composition (r(g) = -.05 to .06), whereas it was positively related to fiber CSA (r(g) = .68). Type IIBw fiber percentage was related to pH at 30 min (r(g) = -.46), pH at 24 h (r(g) = .62), glycolytic potential (r(g) = .31), and lightness of color (r(g) = .55) of longissimus muscle. PMID- 9419986 TI - Effect of the estrogen receptor locus on reproduction and production traits in four commercial pig lines. AB - We investigated the effect of the estrogen receptor (ESR) gene on growth and reproductive traits in four Large White-based commercial pig lines. A total of 9,015 litter records from 4,262 sows genotyped at the ESR locus were analyzed to determine whether ESR influenced total number born (TNB) or number born alive (NBA). Teat number (TN), test ADG, ADFI, feed:gain ratio (F/G), and ultrasonic backfat (BF) were also analyzed to determine effects of ESR. The TNB and NBA were increased per favorable allele of ESR (P < .01) with additive effects of .42 (.31) and .39 (.31) pigs/litter in the first parity (later parities), respectively. Dominance effects were near zero in parity one, but they were .16 and .14 pigs for TNB and NBA, respectively, in later parities (P < .05). A favorable additive pleiotropic effect was detected for BF (P < .001; -.11 mm per copy of the favorable litter size allele). There were no detectable effects on ADG or F/G (P > .10), although ADF was reduced 18 g/d per copy of the favorable litter size allele (P < .05). Average TN was 13.1 for pigs carrying the favorable litter size allele vs 13.2 for noncarriers (P < .05). Marker-assisted selection using ESR is warranted to increase litter size in the Large White-based lines considered here and will be of considerable economic value to pork producers. PMID- 9419987 TI - Effects of prenatal stress on suckling calves. AB - Pregnant Brahman cows (n = 42), bred to either Brahman or Tuli bulls, were randomly assigned to one of three treatments: 1) transported in a stock trailer for 24.2 km, unloaded at a second farm and penned for 1 h, and then returned to the original farm (TRANS); 2) i.v. injection of ACTH, 1 IU/kg BW (ACTH); or 3) walked through the handling facilities (SHAM). Treatments were initiated on d 60 and repeated at 80, 100, 120, and 140 d of gestation. The calves from these cows were subjected to tests to measure their capacity to react to stress. In Test 1, Tuli-sired calves were restrained at 10 and 150 d of age for 3.5 h. In Test 2, Brahman-sired calves were restrained for 3.5 h and given an injection of ACTH (.125 IU ACTH/kg of BW). In Test 3, Test-2 calves were restrained at 180 d of age and hot-iron branded. In Test 4, Test-1 calves were restrained at 180 d of age and given an injection of cortisol (6.7 ng/kg BW) to estimate cortisol clearance rate. During all tests, calves were restrained for 3.5 h, and heart rates were recorded and blood samples were taken at -15, 0, 15, 30, 45, 60, 90, 120, and 180 min. The 10- and 150-d-old TRANS calves maintained greater plasma cortisol in Test 1 (restraint) than the ACTH and SHAM calves (P < .01). The ACTH challenge (Test 2) increased plasma cortisol and ACTH, but cow treatment did not alter the response (P > .4). In response to branding (Test 3), the TRANS, ACTH, and SHAM calves' overall mean plasma cortisol was not affected by treatment (52, 51, and 43 +/- 3 ng/mL, respectively; P > .1), nor was the calves' overall heart rate (91, 94, and 86 +/- 3 beats/min, respectively; P > . 1). In Test 4, TRANS calves cleared plasma of cortisol at a slower rate than did the SHAM calves (P < .01), but not the ACTH calves (261, 374, and 473 +/- 50 mL/min, respectively; P > .1). The TRANS calves had an overall greater heart rate than did the ACTH or the SHAM calves (91, 79, and 77 +/- 2 beats/min, respectively; P < .001). Exposing cows to repeated transportation stress during gestation altered their calf's physiological response to stress, and these alterations could have a profound influence on the calfs ability to adapt to stress, thereby influencing its welfare. Further research should examine the growth, immune function, and reproductive function of prenatally stressed calves to determine whether these changes in plasma cortisol are beneficial or deleterious. PMID- 9419988 TI - Effect of porcine somatotropin administration in young pigs during the growth phase from 10 to 25 kilograms. AB - We conducted two experiments to determine the efficacy of exogenous porcine somatotropin (pST) on enhancing performance during an early phase of growth (10 kg initial BW) when pigs are already growing efficiently and have high rates of lean deposition and low rates of lipid deposition. In Exp. 1, performance was measured on 45 barrows that received one of five daily doses (0, 50, 100, 150 and 200 microg/kg BW) of recombinant pST. In Exp. 2, 27 barrows were used in a slaughter-balance study in which two groups received daily either buffer (control) or 120 microg/kg BW of pST and the third group was slaughtered for initial body composition. In both experiments, pigs received daily i.m. injections of their respective dose for 20 d. The diet was fed for ad libitum consumption and calculated to contain 3.5 Mcal DE/kg, 22.3% CP, and 1.5% lysine. Administration of pST failed to alter overall growth rate or efficiency of gain in either experiment. However, in Exp. 2 pigs treated with pST had increased deposition rates of protein and water but reduced lipid deposition rates. Furthermore, pST treatment resulted in characteristic reductions in plasma urea nitrogen and elevations in glucose and NEFA. Plasma concentrations of insulin and IGF-I were also increased, but pST reduced IGF-II and IGF binding protein-2. Overall, the data demonstrated that very young pigs respond to pST with enhanced lean tissue accretion and metabolic changes, but the response is attenuated compared with previous studies in older growing pigs. PMID- 9419989 TI - Adipose tissue beta-adrenergic and A1 adenosine receptors in suckling pigs. AB - During the first few weeks after birth, major changes occur in porcine adipocyte lipid metabolism. Two of the important receptors controlling lipid metabolism in adipocytes are the beta-adrenergic receptors (betaAR) and the A1 adenosine receptors (A1R). To gain insight into the role of these receptors in modulating neonatal adipocyte lipid metabolism, we measured receptor affinity and number in suckling pigs. Adipose tissue from crossbred (X-Bred) and genetically obese suckling pigs at 0, 3, 10, and 17 d of age was used to prepare crude membranes. The betaAR and A1R number and affinity were measured in membranes by equilibrium saturation binding with radioligands. Obese pigs were smaller than X-Bred pigs (average weight = 1.62 and 2.43 kg for obese and X-Bred, respectively; P < .01). Osmium-fixed adipocytes were larger in obese pigs than in X-Bred pigs (average cell diameter = 34.4 and 30.1 microm for obese and X-Bred, respectively; P < .01). In the obese and X-Bred pigs, the affinity of the betaAR for iodocyanopindolol was greater (lower Kd) at 17 d than at the younger ages (average Kd = 177 pM at 17 d compared with > 330 pM at younger ages; age effect P < .01). The pattern for the betaAR number was complex; the lowest receptor number was at 10 d of age in obese and X-Bred pigs (average number = 41 at 10 d compared with > 65 fmol/mg protein at older and younger ages; age effect P = .03). The higher betaAR Kd and the lower receptor number in younger animals suggest less regulation by physiologic concentrations of epinephrine and norepinephrine. This would allow greater anabolic lipid metabolism to proceed during the neonatal period, when adipocytes increase four- to sixfold in volume. There were no measurable A1R at any of these early ages; thus, adenosine control mechanisms to counteract the betaAR and provide negative controls to lipid accretion are not operable in suckling pigs. PMID- 9419990 TI - Influence of sample orientation on prediction of fresh ham lean content by electromagnetic scanning. AB - To evaluate the effect of orientation of hams during electromagnetic scanning on the estimation of dissected lean content, hams were scanned horizontally, posterior first (POS) or dorsal first (DOR), and vertically, medial side (aitchbone) first (MED; standing on the butt face). Weight and percentage of dissected lean were estimated using scan peak for each orientation, ham weight, and fat thickness. The mean scan peak for the MED orientation was approximately twice as great as peaks for the POS and DOR orientations, which suggests that this orientation may offer greater predictive accuracy by reducing the signal to noise ratio. Results, however, indicated that all orientations were equally effective at predicting lean weight and percentage, with R2 values of .95 and .75 and root mean square errors of .21 kg and 2.6%, respectively. PMID- 9419991 TI - Phytase supplementation of low-phosphorus growing-finishing pig diets improves performance, phosphorus digestibility, and bone mineralization and reduces phosphorus excretion. AB - Two experiments using 413 crossbred growing-finishing pigs were conducted to assess the use of a commercial microbial phytase (Natuphos) in corn-soybean meal diets to improve phytate P bioavailability and thus reduce inorganic P supplementation and fecal P excretion. In Exp. 1 (n = 189), the following diets were used: 1) .50/.40% total P, respectively, for grower and finisher phases, and no phytase; 2) .40/.35% P and no phytase; 3) diet 2 plus 250 U phytase/kg; and 4) diet 2 plus 500 U phytase/ kg. The total Ca level was .58/.48% for diet 1 and .53/.43% Ca for diets 2, 3, and 4 in the grower and finisher phases, respectively. Feeding the low-P diet without supplemental phytase resulted in an overall 18% reduction in ADG (P < .05), 15% reduction in ADFI (P < .05), and 3% poorer feed efficiency (P < .08). Adding 250 to 500 U phytase/kg to the low-P diet restored ADG, ADFI, and feed conversion to levels not significantly different from and within 96% of that observed for pigs fed the adequate-P diet. The overall apparent digestibility of P was linearly (P < .01) improved with addition of 250 and 500 U phytase/kg to the low-P diet, but Ca and DM digestibilities were not affected by phytase or P level. In Exp. 2 (n = 224) the following diets were used: 1) .38/.33% total P, respectively, for grower and finisher phases, and no phytase; 2) .42/.37% P and no phytase; 3) .46/.41% P and no phytase; 4) diet 1 plus 167 U/kg phytase; 5) diet 1 plus 333 U/kg phytase; and 6) diet 1 plus 500 U/kg phytase. All diets contained .41/.36% Ca for grower and finisher phases, respectively. Pigs fed the low-P control diet grew slower (P < .01) and less efficiently (P < .10) than pigs fed diets with added P or phytase. With increasing levels of supplemental phytase or P there was a linear increase (P < .01) in ADG, digestibility of P, and digested P and a quadratic improvement (P < .05) in feed efficiency. Tenth rib mineralization based on shear force and ash were linearly increased (P < .08 to .001) as phytase or P was added to the low-P diet. There were generally no effects of P or phytase level on carcass quality. Using prediction equations derived from the response traits of ADG and P digestibility in Exp. 1 and ADG, P digestibility, and bone shear force in Exp. 2 to added phytase or P, we estimated that 500 U phytase released an amount of phytate P that was approximately equivalent to .87 to .96 g of P from dicalcium monocalcium phosphate supplements. Fecal P excretion was estimated to be reduced 21.5%. PMID- 9419992 TI - Nutritional evaluation of biologically treated white kidney beans (Phaseolus vulgaris L.) in pigs: ileal and amino acid digestibility. AB - We studied the effect of feeding young growing pigs a semisynthetic diet containing 7.5% white kidney beans-germinated (GB), pancreatin treated (PTB), or untreated (raw beans RB)--on protein and amino acid (AA) digestibilities at the terminal ileum. Eleven castrated male pigs (12.2 kg live weight) fitted with a post-valve T-cecal cannula and two blood catheters were used. The 15N-isotope dilution method was used to determine the amount of endogenous protein passing the terminal ileum and the true ileal protein digestibility. Ileal crude protein losses in pigs fed the RB, GB, and PTB diets were 51.9, 27.4, and 51.1 g/kg of DMI, respectively. The total amounts of AA passing the terminal ileum of the pigs fed the RB, GB, and PTB diets were 48.6, 21.4, and 42.2 g/kg DMI, respectively. The apparent ileal crude protein and AA digestibilities of the RB, GB, and PTB diets were 74, 87, and 75% and 76, 89, and 78%, respectively. True ileal protein digestibilities were 88, 93, and 93% for the RB, GB, and PTB diets, respectively. On the basis of this research, germination of white kidney beans improves the digestion of protein by decreasing the content of bean antinutritional factors and increasing the bean true ileal protein digestibility. PMID- 9419993 TI - Relative bioavailability of supplemental inorganic zinc sources for chicks. AB - Three experiments were conducted to investigate the relative bioavailability of reagent-grade (RG) and feed grade (FG) Zn sources for 1-d-old broiler chicks. In Exp. 1, 13 treatments included a basal corn-soybean meal diet (63 ppm Zn) or the basal diet supplemented with 400, 800, or 1,200 ppm Zn from RG sulfate, basic carbonate, oxide, or metal and fed for 20 d. Using multiple regression slope ratios with Zn sulfate set at 100%, bioavailability estimates were 78, 77, and 46% for carbonate, oxide, and metal, respectively. In Exp. 2, chicks were allotted randomly to 16 treatments that included a basal corn-soybean meal diet (75 ppm Zn) or basal diet supplemented with 300, 600, or 900 ppm Zn as either RG sulfate, FG sulfate-A, FG sulfate-B, FG oxide-A, or FG oxide-B and fed for 21 d. Multiple linear regression slope ratios gave relative estimates of 99, 81, 78, and 54% for sulfate-A, sulfate-B, oxide-A, and oxide-B sources, respectively, with RG sulfate set at 100%. In Exp. 3, chicks were fed a basal corn-soybean meal diet (35 ppm Zn) or the basal diet supplemented with 40, 80, or 120 ppm Zn from RG Zn sulfate, FG sulfate, or FG oxide and fed for 20 d. Multiple regression slope ratios with RG sulfate set at 100% gave relative bioavailability estimates of 94 and 74% for the FG sulfate and oxide, respectively. Bioavailability estimates were similar when Zn was supplemented to diets at high or low concentrations. PMID- 9419994 TI - Estimating endogenous amino acid flows at the terminal ileum and true ileal amino acid digestibilities in feedstuffs for growing pigs using the homoarginine method. AB - True ileal lysine digestibilities were determined using the homoarginine (HA) method in casein-, barley-, canola meal-, and barley-canola meal-based diets fed to growing pigs. Four Yorkshire barrows (25 to 49 kg BW) equipped with ileal T cannulas were fed one of the diets according to a 4 x 4 Latin square design. All diets, except for the barley diet, were formulated to be similar in DE:CP ratio. Ileal digesta were collected continuously for 24 h on d 7 and 9 of each experimental period for determining apparent and true ileal digestibilities, respectively. True ileal lysine digestibility and endogenous flows were determined by feeding diets in which 50% of the protein-containing ingredients was guanidinated; the conversion of lysine to HA allows for direct determination of true lysine digestibilities. The apparent ileal CP and amino acid digestibilities were higher (P < .05) in the casein diet than in the barley, canola meal, and barley-canola meal diets. The CP digestibility was higher (P < .05) in the barley-canola meal diet than in the barley and canola meal diets. Endogenous lysine losses were influenced by diet (P < .05) and ranged between 586 and 1,429 mg/kg DM intake. True ileal lysine digestibilities in barley, canola meal, and barley-canola meal diets were similar (P > .05) and lower (P < .05) than in the casein diet. The true ileal amino acid digestibilities were estimated for other amino acids. Unlike apparent digestibilities, true digestibilities seemed additive in the barley-canola meal mixture. PMID- 9419995 TI - Evaluation of the interrelationships among lactose and protein sources in diets for segregated early-weaned pigs. AB - We conducted two experiments to evaluate the interactions among lactose and protein sources in diets for segregated early-weaned pigs. In Exp. 1, 360 barrows (initially 5.3 kg and 19 +/- 2 d of age) were fed diets containing crystalline lactose (0, 20, and 40%), spray-dried animal plasma (0 and 7.5%), and soybean meal (0 and 20%) in a 3 x 2 x 2 factorial arrangement. We used a blend of select menhaden fish meal and casein to replace the lysine provided by soybean meal or animal plasma. Diets contained 1.7% total lysine and were fed from d 0 to 14 after weaning. Pigs were fed a common diet from d 14 to 34. From d 0 to 14 after weaning, ADG and ADFI increased with increasing dietary lactose when the diet contained soybean meal but decreased when soybean meal was not in the diet (lactose x soybean meal, P < .05 and .10, respectively). Pigs fed animal plasma had increased (P < .05) ADG and ADFI from d 0 to 14 but decreased (P < .05) ADG from d 14 to 34. In Exp. 2, 324 barrows (initially 3.7 kg and 10 +/- 2 d of age) were fed diets from d 0 to 10 similar to those used in Exp. 1 with the exception that extruded soy protein concentrate replaced the lysine provided by soybean meal or animal plasma. From d 0 to 10 after weaning, increasing lactose improved (linear, P < .05) ADG and ADFI, and pigs fed animal plasma had higher ADFI (P < .05). In conclusion, soybean meal had no negative effect on ADG; however, animal plasma and lactose increased ADG and ADFI for pigs weaned between 10 and 19 d of age. PMID- 9419996 TI - Effects of substituting deproteinized whey and(or) crystalline lactose for dried whey on weanling pig performance. AB - We conducted two trials to determine the effects of replacing the lactose provided by spray-dried, edible-grade whey with edible-grade deproteinized whey or crystalline lactose on pig performance. In Exp. 1, 180 weanling pigs (initially 4.1 kg and 22 +/- 4 d of age) were allotted randomly to dietary treatments containing 18% lactose supplied by 1) 25% dried whey, 2) 12.5% dried whey and 9% crystalline lactose, 3) 18% crystalline lactose, 4) 12.5% dried whey and 10.9% deproteinized whey, or 5) 21.7% deproteinized whey. Casein was used to replace the lysine provided by dried whey in diets containing lactose and deproteinized whey. From d 0 to 14 after weaning, no differences (P > .10) were observed in ADG or ADFI. Pigs fed diets containing 18% crystalline lactose or 21% deproteinized whey had a higher (P < .05) gain:feed ratio (G/F) than did pigs fed diets containing 25% dried whey or 12.5% dried whey and 9% lactose. In addition, pigs fed diets containing 21% deproteinized whey had increased G/F compared to pigs fed the diet containing 10.9% deproteinized whey and 12.5% dried whey. In Exp. 2, 344 pigs (initially 4.4 kg and 14 +/- 2 d of age) were fed dietary treatments based on four sources of crystalline lactose replacing the lactose provided by dried whey in the positive control diet (20% dried whey). In addition, a negative control diet was formulated with 7.2% crystalline lactose. Casein was used to replace the lysine provided by dried whey. From d 0 to 14 after weaning, no differences (P > . 10) were observed in performance. However, pigs initially fed the positive control diet subsequently (d 14 to 28) consumed more feed than pigs fed the negative control diet. These results indicate that edible-grade deproteinized whey and crystalline lactose can replace the lactose provided by high-quality dried whey without affecting pig performance. PMID- 9419998 TI - Regulatory roles of high-density and low-density lipoproteins in cellular proliferation and secretion of progesterone and insulin-like growth factor I by enriched cultures of bovine small and large luteal cells. AB - We tested the hypotheses that bovine high-density (HDL) and low-density (LDL) lipoproteins differentially influence cellular proliferation and progesterone and IGF-I production by bovine small and large luteal cells. Unit gravity sedimentation was used to produce enriched cultures of small (> 95% pure) and large (75 to 90% pure) luteal cells from corpora lutea (CL) on d 4 and 10 of the estrous cycle. Addition of LDL, HDL, or both resulted in the maintenance of higher (P < .05) numbers of 3beta-hydroxysteroid dehydrogenase (HSD)-positive small and large cells in culture and produced a marked proliferation of 3beta-HSD negative small luteal cells compared to control medium. Low-density lipoprotein and HDL each stimulated greater (P < .01) progesterone secretion in enriched large cell cultures on both days of the cycle, and by small luteal cells on d 10, compared to the control. Together, LDL and HDL maximized this response. Lipoproteins markedly stimulated (P < .01) the secretion of IGF-I by bovine large luteal cells, and secretion was greater (P < .05) by cells from d 10 CL compared to d 4 CL. Results suggest that the actions of lipoproteins in bovine luteal cells extend beyond their widely recognized roles in steroidogenesis and include remarkable effects on cellular proliferation and IGF-I secretion. Type of lipoprotein (LDL vs HDL) did not have differential effects on any of the variables measured. PMID- 9419997 TI - Effects of locoweed (Oxytropis sericea) on growth, reproduction, and serum hormone profiles in young rams. AB - Sixteen ram lambs (5 mo old, average BW = 49 +/- 1.4 kg) received a control diet (50% concentrate, 13.1% CP) or a diet containing 7, 14, or 21% locoweed (LW, DM basis). Rams were housed in individual pens (2.5 x 3.5 m) in an enclosed facility (14 h light, 10 h dark) with free access to feed and water. After 35 d of LW feeding, rams fed the 21% LW diet ate 1.3 to 1.6 kg/d, whereas rams fed the other three diets ate 1.6 to 2.0 kg/d. On d 35, BW (mean +/- SE) were 58.3, 57.2, 57.2, and 55.4 (+/- 1.5 kg/d) for ram lambs receiving 0, 7, 14, and 21% LW, respectively (P > .20). An inverse relationship for BW (P < .10) and gain (P = .04) was observed 1 mo after LW feeding ended relative to amount of LW in the diet. Serum alkaline phosphatase increased with increasing LW (P < .05), and thyroxine concentration was decreased (P < .05) by LW ingestion. On d 35, serum LH averaged 6.8, 9.9, 11.6, and 9.9 (+/- 1.8) ng/mL (P = .56), whereas testosterone averaged 3.5, 2.6, 3.0, and 1.6 (+/- .5) ng/mL (linear effect of LW, P = .05) over a 5-h period after GnRH injection (i.m., 50 microg) in lambs fed 0, 7, 14, and 21% locoweed, respectively. Semen volume, sperm motility, sperm cell concentration, and percentage of abnormal cells did not differ (P > .50) on d 35. One month after the end of LW feeding, a linear (P = .06) decrease in sperm motility and scrotal circumference was observed relative to percentage of prior dietary LW. These data suggest that LW exerts adverse effects on the testes of young rams that may not be evident until several weeks after LW consumption ends. PMID- 9419999 TI - Porcine follicle-stimulating hormone treatment of gilts during an altrenogest synchronized follicular phase: effects on follicle growth, hormone secretion, ovulation, and fertilization. AB - Porcine FSH (SUPER OV), containing .03% LH activity, and equine chorionic gonadotropin (eCG) were administered during an altrenogest-synchronized follicular phase to determine their effects on follicle development, estrus, ovulation, and fertilization. Treatments were made by i.m. injection starting on d 1 (24 h after the last feeding of altrenogest): 1) saline, once, n = 14; 2) eCG (1,200 to 1,500 IU) once, n = 32; 3) FSH 14 (n = 2) or 21 (n = 6) NIH-FSH-S1 units/100 kg BW, divided among six injections at 12-h intervals (FSH14/21); 4) FSH, 28 NIH-FSH-S1 units/100 kg BW, divided among six injections at 12-h intervals, n = 12; and 5) FSH, 28 NIH-FSH-S1 units/100 kg BW and 100 IU hCG, two or six injections at 12-h intervals (FSH28+hCG), n = 13. Gilts were injected with 750 IU of hCG on d 5 to ensure ovulation. Twenty-eight eCG- and FSH-injected gilts (n = 6, 8, and 11 on treatments 3, 4, and 5, respectively) were bred and laparotomized on d 7 to recover ova and record ovulation rate. The mean number of ovulations and large (6- to 10-mm) follicles, respectively, on d 7 were as follows: saline (17, .7), eCG (43, .9), FSH14/21 (15, .6), FSH28 (12, 16), and FSH28+hCG (32, 21). Plasma FSH concentrations were at least threefold higher (P < .05) in gilts treated with FSH than in gilts not treated with FSH. The percentage in estrus was higher (P < .05) for saline- and eCG-treated gilts (100 and 87%, respectively) than for FSH-treated gilts (53%). Proportion of FSH28+hCG-treated gilts with fertilized ova (27%) was lower than for other groups (79 to 100%). In summary, the 3-d high dose FSH treatment (FSH28 and FSH28+hCG) during an altrenogest-synchronized follicular phase increased the number of potentially ovulatory follicles, but this potential benefit was not realized because many follicles failed to ovulate. The co-injection of low doses of hCG (FSH28+hCG) increased the ovulation rate and estradiol secretion but reduced ova recovery and fertilization rate compared with eCG and the other FSH treatments. PMID- 9420000 TI - Concentration of tissue inhibitor of metalloproteinases (TIMP)-1 in ovine follicular fluid and serum. AB - Tissue inhibitor of metalloproteinases (TIMP)-1 mRNA and protein localize within granulosal cells of post-gonadotropin-surge follicles and luteal tissue in ewes. Our objectives were to test the hypotheses that 1) follicular fluid concentration of TIMP-1 increases following a gonadotropin surge induced by LHRH agonist (Exp. 1) and 2) luteal status affects peripheral serum concentration of TIMP-1 (Exp. 2 and 3). In Exp. 1, the concentration of TIMP-1 within antral fluid from post surge follicles (28.7 +/- 6.65 microg/mL) was greater (P < .02) than from pre surge follicles (2.37 +/- 2.47 microg/mL). In Exp. 2, serum concentration of TIMP 1 did not differ among d 0 to 6 (1.27 +/- .55 microg/mL) of the estrous cycle or among periods of luteal maintenance (1.29 +/- .06 microg/mL), spontaneous luteal regression (1.19 +/- .09 microg/mL), or luteal development (1.22 +/- .08 microg/mL). However, serum concentration of TIMP-1 was greater ( P < .001) during the period of luteal maintenance (1.14 +/- .04 microg/mL) than during PGF2alpha induced luteolysis (d 26; .85 +/- .06 microg/mL) and induced luteal absence (d 27 to 33; .95 +/- .05 microg/mL). In Exp. 3, ewes (n = 14) were bled daily from d 1 to 19 (d 0 = estrus) and at 12-min intervals for 6 h on d 3, 10, and 17. Although concentration of TIMP-1 varied considerably within and among ewes, mean concentration of TIMP-1 per ewe per day increased ( P < .05) from d 3 to 17. These data indicate that follicular fluid concentration of TIMP-1 increases following a gonadotropin surge, but the contribution of ovarian derived TIMP-1 to peripheral serum concentration is negligible. PMID- 9420001 TI - Continuous profiles and within-day variations of metabolites and hormones in cows fed diets varying in alimentary supplies before short-term feed deprivation. AB - We investigated continuous profiles and within-day variations of some metabolites and hormones in four nonpregnant, nonlactating cows fed hay-based diets in two equal meals. Diets supplied either too much or too little N (approximately 1.3 or .8 times the maintenance requirements) and NE1 (approximately 1.2 or .8 times). Continuous collection of ruminal liquor, blood, and urine samples was performed for 42 consecutive hours, the last 16 h covering a period without feed. For twice daily feeding, nitrogenous and energetic underfeeding decreased average ruminal propionate and the insulin:growth hormone ratio. However, only the energetic underfeeding increased plasma 3-methylhistidine and urinary excretion of 3 methylhistidine, and decreased body weight and ruminal acetate, butyrate, and total VFA. Conversely, only the nitrogenous underfeeding decreased glycemia. Whatever the dietary level, the 42-h patterns of metabolites and hormones were mainly affected by the time from the last meal. An energy deficit progressively took place during feed deprivation and the nocturnal interprandial period but not during the diurnal interprandial period. During the feed deprivation and nocturnal periods, glycemia was maintained despite a shortage in ruminal propionate. We conclude that in twice-daily fed cattle 1) the dietary supply of energy is the main trigger for an energy deficit and the subsequent muscle protein mobilization; 2) the nocturnal interprandial period may be considered as short-term feed deprivation; 3) the diurnal patterns of metabolites and hormones are not affected by the nitrogenous and(or) energetic supplies of the diet. PMID- 9420002 TI - Protein requirements of growing steers limit-fed corn-based diets. AB - In Exp. 1, six steers (254 kg) were used in a 6 x 4 incomplete Latin square to determine the effects of solvent-extracted soybean meal alone or in combination with rumen-protected methionine and lysine on N balance in steers limit-fed a high-corn diet to gain 1.1 kg/d. The basal diet contained (DM basis) 80% rolled corn, 15% alfalfa, and .9% urea (13.9% CP), and 2 or 4% soybean meal replaced corn to give CP concentrations of 14.8 and 15.6%, respectively. Each diet was fed with and without 5 g/d of Smartamine-ML (.75 and 2.0 g of rumen-protected methionine and lysine, respectively). Nitrogen retention increased linearly (P = .09) with level of soybean meal. Rumen-protected methionine and lysine had no effect on N balance. In Exp. 2, seven steers (233 kg) were used in a 7 x 4 incomplete Latin square experiment to investigate optimal levels and sources of CP for steers limit-fed to gain 1 kg/d. Treatments included a negative-control diet (urea; 11.7% CP) and six diets containing either 13.5, 15.4, or 17.2% CP with either solvent-extracted or expeller-processed soybean meal. Diets provided 75, 87.5, 100, or 112.5% of estimated CP requirement for a gain of 1 kg/d. The basal diet contained 83% rolled corn, 15% alfalfa, and .2% urea. Nitrogen retention increased linearly (P = .006) with soybean meal addition, and no differences were observed between CP sources. The CP system underpredicted the protein requirements of limit-fed steers under our conditions. PMID- 9420003 TI - Plasma insulin, metabolite concentrations, and carcass characteristics of Japanese Black, Japanese Brown, and Holstein steers. AB - To characterize some of the physiological features of Japanese beef breeds, plasma concentrations of insulin and metabolites and carcass composition were measured in five Japanese Black, five Japanese Brown, and four Holstein steers (6.2 mo; 164 kg). The steers were raised under typical feeding conditions in Japan until they were slaughtered at 600 to 700 kg BW. Blood samples were collected at 8 mo of age (average BW, 194 kg) and at 300, 400, 500, and 600 kg BW. Plasma insulin concentrations increased with BW in all three breeds and were greater (P < .05) in Japanese Blacks than in the Japanese Browns or Holsteins at 400 and 600 kg BW. The Japanese Blacks exhibited lower (P < .05) plasma glucose levels at 300, 400, and 600 kg BW compared with Holsteins. Regardless of the breed, plasma urea nitrogen (PUN) concentrations increased with BW. The two Japanese breeds had greater (P < .05) PUN levels than Holsteins at 300 and 600 kg BW. Total cholesterol and phospholipid concentrations tended to decrease above 300 kg BW in the Holsteins; however, the concentrations of both metabolites were elevated in the steers of Japanese breeds at 500 and 600 kg BW (P < .05). Breed did not affect the plasma concentrations of albumin, triglycerides, and NEFA. The Japanese breeds had higher (P < .01) dressing percentage, greater (P < .05) carcass fat proportion, and a lower proportion of carcass bone (P < .01) than the Holsteins. These results indicate that there are breed differences in plasma levels of insulin and certain metabolites and carcass composition among Japanese breeds and Holstein. PMID- 9420004 TI - Limiting amino acids in meat and bone and poultry by-product meals. AB - In situ, digestion, and growth studies were conducted to evaluate four meat and bone meals and six poultry by-product meals as sources of escape protein and to predict the first-limiting amino acid for growing calves. Escape protein values, determined by 12-h in situ incubation, ranged from 41.7 to 51.0% of CP for meat and bone meals; poultry by-product meals ranged from 32.0 to 39.8%. True protein digestion in the gastrointestinal tract of lambs differed among protein sources (P < .05), ranging from 79 to 95%. In each of three growth trials, 60 steers (258 +/- 24, 241 +/- 23, and 230 +/- 16 kg for Trials 1, 2, and 3, respectively) were supplemented with 4 of the 10 protein sources along with a urea supplement. Protein sources were fed at 30, 40, 50, and 60% of the supplemental CP, with urea supplying the remainder. Protein efficiency differed among treatments ( P < .10), ranging from .61 to 1.55. Amino acid composition was determined for each protein source, and the individual metabolizable amino acids were regressed on the protein efficiency values. Escape protein values were correlated (R2 = .75) with protein efficiency but had a negative slope. Metabolizable methionine was the only amino acid moderately correlated (R2 = .40, slope = 1.9) to protein efficiency, whereas other amino acids either correlated poorly or had negative slopes. These data indicate that the protein value of meat and bone meal and poultry by-product meal is limited by the amount of metabolizable methionine they contain. PMID- 9420005 TI - Addition of ruminal escape methionine and lysine to meat and bone meal. AB - A growth study was conducted to determine the effects of adding ruminal escape methionine and lysine to meat and bone meal (MBM). A basal diet of 44% sorghum silage, 44% corncobs, and 12% supplement (DM basis) was individually fed to 60 crossbred steers (234 +/- 14 kg). Supplements contained either urea, MBM, MBM plus protected methionine (MBM + M), or MBM plus protected methionine and lysine. Protein sources were fed to supply 30, 40, 50, and 60% of the supplemental CP, with urea supplying the remainder. Protein efficiency, calculated as gain above the urea control vs natural protein intake using the slope ratio technique, was used to evaluate the protein sources. The most efficiently used protein source was MBM + M, which was greater than MBM alone (P < .10). Meat and bone meal plus protected methionine and lysine had a protein efficiency similar to MBM + M (P > .30), indicating that lysine was not limiting. True protein digestibility of MBM in the gastrointestinal tract of lambs was determined to be 86.1%. In situ analysis performed by 12-h ruminal incubation of MBM determined the escape CP to be 53.0% of CP. Amino acid analysis was conducted to compare supplies to requirements for live animal gain. The urea control failed to meet the metabolizable protein requirement. Feeding MBM to provide additional metabolizable protein failed to provide an adequate amount of the essential amino acid methionine, which was first-limiting. These data indicate that protein efficiency of MBM can be enhanced by the addition of ruminal escape methionine. PMID- 9420006 TI - Absorption of amino acids from the intestine and their net flux across the mesenteric- and portal-drained viscera of lambs. AB - Experiments were conducted to compare the rates of apparent absorption (disappearance) of individual essential amino acids (EAA) from the small intestine with their net fluxes across the mesenteric- (MDV) and portal- (PDV) drained viscera in sheep given a pelleted alfalfa diet at two levels of intake. Disappearances of individual EAA across the region of the small intestine drained by the mesenteric arcade (jejunum to ileum) were similar to those across the whole of the small intestine (duodenum to ileum). The net MDV flux of each EAA was similar to its rate of disappearance, but, with the exception of threonine on the low intake level, the net PDV flux was lower (P < .05). Increasing the intake of alfalfa from 800 to 1,200 g/d increased the rate of disappearance of individual EAA between the duodenum and ileum by .56 (range .43 to .65) and between the jejunum and ileum by .51 (range .45 to .60). The MDV and PDV blood flows increased by .35 and .39, respectively, and, with the exception of valine, net fluxes of individual EAA increased by .39 (range .20 to .50) across the MDV and by .44 (range .21 to .71) across the PDV. When net fluxes across the MDV and PDV were measured simultaneously, the ratio of PDV: MDV flux for each EAA was less than (P < .05) unity (mean .61, range .55 to .69), even though all MDV blood enters the PDV, contributing approximately .45 of the total portal flow. This observation suggests that, in regions of the tract not drained by the MDV, extraction of arterial blood EAA for tissue and secretory protein synthesis must exceed the release of protein degradation products into the venous drainage. The results are discussed in terms of endogenous protein secretions into the lumen of the tract anterior to the small intestine. PMID- 9420007 TI - Quantification of circulating peptides and assessment of peptide uptake across the gastrointestinal tract of sheep. AB - Gastrointestinal absorption of peptides was examined in sheep fed a forage-based diet. Peptide concentrations were determined in arterial, portal, and mesenteric blood and plasma by quantification of amino acid concentrations before and after acid hydrolysis of samples that had been first deproteinized then subjected to Sephadex G-15 gel-filtration to remove residual protein. In contrast to other studies of ruminants, peptide concentrations for individual amino acids were lower than for the corresponding free amino acids with peptide (expressed as a proportion of total nonprotein amino acid) representing not more than .25 to .3 of total amino acid. Peptide concentrations in arterial, mesenteric, and portal blood and plasma were similar, indicating that on this diet there was no net uptake of peptides from the small intestine (mesenteric-drained viscera, MDV) or the whole tract (portal-drained viscera, PDV). Increasing the intake of alfalfa pellets from 800 to 1,200 g/d, while increasing the absorption and net flux across the MDV and PDV of free amino acids, had no effect on peptide absorption. Preparation of blood and plasma samples for peptide analysis with methods used in studies in which substantial peptide absorption has been reported indicated no net MDV or PDV flux of peptide. Such conflicting data on the extent of gastrointestinal peptide flux are discussed in the context of methodological differences and the importance of diet and physiological state of the animal. PMID- 9420008 TI - Poly(A)+ RNA from sheep omasal epithelium induces expression of a peptide transport protein(s) in Xenopus laevis oocytes. AB - To verify research from this laboratory indicating that sheep omasal epithelium contains mRNA encoding for a peptide transporter(s) and to determine di- to octapeptide transport capability, we injected poly(A)+ RNA isolated from sheep omasal epithelium into Xenopus laevis oocytes. Poly(A)+ RNA was functionally expressed in Xenopus oocytes 4 to 7 d after injection. Peptide (5 di-, 10 tri-, 6 tetra-, 2 penta-, 1 hexa-, 1 hepta-, and 1 octapeptide) transport capability was measured by impaling oocytes with a microelectrode to monitor membrane potential (Vm). Oocytes were maintained in pH 5.5 buffer. Peptide transport was identified as being expressed when, in the presence of a buffered peptide substrate (1 mM), the oocyte membrane showed persistent depolarization (a more positive Vm). In the absence of peptide transport, the membrane became depolarized with the addition of buffered substrate, but it rapidly repolarized to the resting potential. Peptide transport was expressed for some di-, tri-, and tetrapeptides. Measured depolarization ranged from 9.6 mV to 42.1 mV. Larger peptides were not transported by the oocytes. When transport expression was measured with the substrates in a pH 7.5 buffer, no transport occurred, indicating that transport was dependent on a proton gradient. Thus, sheep omasal epithelium contains mRNA that codes for a protein(s) capable of proton-dependent di-, tri-, and tetrapeptide transport. Results from the present study provide further evidence that absorption of peptides from the ruminant stomach is possible. PMID- 9420009 TI - Comparison of hindquarter metabolite uptakes in Belgian Blue double-muscled bulls at maintenance or during fattening. AB - Metabolism of muscle growth in the hindquarter was investigated by the arterio venous difference (AVD) technique in Belgian Blue double-muscled type bulls at maintenance or at fattening. The bulls were fitted with an aortic ultrasonic blood flow probe and with catheters in the aorta and vena cava. They were offered a diet allowing for maintenance (MP) during a period of 15 d, at the end of which measurements were made over 3 d. Bulls were then given a fattening diet (FP) and the measurements were repeated. Arterial blood flow was approximately 1 L/min greater when the bulls were standing than when lying. Blood flow was 2 L/min higher during FP than during MP. The AVD and uptake of glucose were maximal at 1400 and 1600. Uptake of alpha-amino nitrogen decreased immediately after a meal. The increase in glucose from MP to FP fitted very well with the calculated energy needs for muscle growth. The AVD and uptake of alpha-amino nitrogen, total amino acids, and total nonessential amino acids were negative during MP and positive and significantly higher during FP. There was also a significant increase in AVD and uptake of essential and branched-chain amino acids when the bulls were changed from MP to FP. When changing from maintenance to fattening, the incremental glucose and amino acid hindquarter uptake provided energy and supply for muscle protein accretion, respectively. The level of alanine transamination was also sharply reduced. PMID- 9420010 TI - Neutral detergent fiber disappearance and gas and volatile fatty acid production during the in vitro fermentation of six forages. AB - Samples of unfractionated forage and isolated NDF from six forages were fermented in vitro, and NDF disappearance and gas and VFA production were measured over time. Rates based on each of these data sets were calculated using a one-pool logistic model. The rates of NDF disappearance and gas and VFA production did not differ within each forage. Gas and VFA production were linearly related to NDF digestion. Gas yield was .35 mL/mg (r2 = .92) of NDF digested for the isolated NDF. The amount of total VFA produced per milligram of NDF digested was more variable than gas (r2 = .72), with a slope of .01 mmol VFA/mg of NDF digested. The relationship between gas and VFA production was linear (mean slope of 1.43 mmol gas/mmol VFA, r2 = .69). The ratios of end products (gas and VFA) to NDF digestion and the ratio of acetate:propionate were variable during the first 8 h of fermentation but changed little after this time. Changes in the acetate: propionate ratio explained 23% of the variation in gas produced per millimole of total VFA detected. PMID- 9420011 TI - The effects of amount of whole barley, barley bulk density, and form of roughage on feedlot lamb performance, carcass characteristics, and digesta kinetics. AB - We conducted two feedlot trials and one metabolism trial to evaluate the effect of barley level, barley bulk density, and physical form of roughage on lamb growth performance and digesta kinetics. Level of whole barley (50, 70, 90%) and type of roughage (chopped or pelleted alfalfa) were evaluated in Trial 1 (50 d period). Trial 2 (50 d) evaluated barley bulk density (heavy = 671 and light = 607 kg/m3), form of roughage (pelleted or chopped alfalfa), and level of barley (80 or 40%). The influence of treatments used in Trial 2 on digesta kinetics was evaluated in Trial 3. Gain:feed increased and DMI decreased (P < .10) linearly with increasing level of barley, and ADG and DMI were greater (P < . 10) for lambs fed pelleted vs chopped alfalfa in Trial 1. The 70% barley diet produced the highest yield grade and kidney-pelvic fat and the lowest leg score among barley levels (P < .10). Lambs fed pelleted alfalfa had heavier carcasses and a thicker body wall than lambs fed chopped alfalfa (P < .02). In Trial 2, DMI was less and gain:feed greater (P < .01) for lambs fed the heavy barley than for lambs fed the light barley and for the 80% barley diet compared to the 40% barley diet. Lambs fed pelleted alfalfa had greater dressing percentages than lambs fed chopped alfalfa. Backfat and body wall thickness were greater (P < .10) for lambs fed the 80% barley diet than for those fed the 40% barley diet. In Trial 3, retention time of barley was greater (P < .10) for lambs fed light rather than heavy barley, and retention time of alfalfa was greater (P < .10) for lambs fed chopped compared with pelleted alfalfa. Acetate:propionate ratio was greater (P < .10) for lambs fed light vs heavy barley and lambs fed the 40 vs 80% barley diets. Ruminal pH was lower (P = .05) and in situ barley digestion greater (P = .03) over time in lambs fed the 80% barley diet than in lambs fed the 40% barley diet. Feedlot lamb ADG was not always greatest with high levels of barley; however, gain:feed improved at the higher barley levels. The higher barley levels seemed to result in fatter lambs. PMID- 9420012 TI - Linkage mapping of porcine interleukin 6 (IL6). PMID- 9420013 TI - Nucleotide sequence of porcine OTCase cDNA. PMID- 9420014 TI - Comment on the need for thoroughness in literature searches. PMID- 9420015 TI - Chemotherapy of colorectal cancer: history and new themes. AB - Since the clinical introduction of 5-fluorouracil (5-FU) in 1958, improvements in the treatment of advanced colorectal cancer have been modest. However, improvements in response rates have been demonstrated when 5-FU is administered in conjunction with leucovorin, and when methotrexate or trimetrexate is administered preceding 5-FU, indicating that higher response rates could be achieved by biomodulating the activity of 5-FU. Thus, significant emphasis has been placed on designing more effective 5-FU-based combination regimens. Novel agents, including the thymidylate synthase inhibitor raltritrexed and the topoisomerase I inhibitor irinotecan, also have demonstrated activity in colorectal cancer. Other new approaches include the administration of oral 5-FU prodrugs. The development of novel agents, new therapeutic approaches, and the refinement of existing agents and regimens in the clinic will likely improve response rates and, ultimately, patient survival. The history, current treatment options, and future opportunities for advances in chemotherapy for the treatment of colorectal cancer are discussed. PMID- 9420016 TI - Systemic treatment options in advanced colorectal cancer: perspectives on combination 5-fluorouracil plus leucovorin. AB - A variety of 5-fluorouracil (5-FU)- based chemotherapy regimens have been investigated in colorectal cancer patients in randomized trials over the past decade. The standard regimen for treatment of colorectal cancer is combination 5 FU plus leucovorin (LV). The results from 12 randomized trials indicate that 5 FU/LV is more active than single-agent 5-FU (25% v 14% of evaluable patients); however, median survival was unchanged (12.2 months v 11.4 months, respectively). Furthermore, the weekly and monthly schedules of 5-FU/LV are therapeutically equivalent, although the spectrum of toxicity differs. On the monthly schedule, a LV dose of 200 mg/m2 appears to have no advantage over 20 mg/m2; however, on the weekly schedule, high-dose LV appeared to be slightly more effective than low dose LV (2-hour infusion) (25% v 18%) at the cost of a higher incidence of severe diarrhea (26% v 14%). Furthermore, similar response rates are observed with the racemic commercial formulation of LV and the pure (I-LV) active stereoisomer. Other modulatory strategies that appear to produce higher response rates than single-agent intravenous push 5-FU include sequential methotrexate/5-FU (19% v 10%) and continuous infusion schedules (22% v 14%). Although 5-FU-modulated strategies improve response rates over those observed with single-agent 5-FU, median survival in multi-institutional trials unfortunately has not generally exceeded 12 months. The mechanism of action, clinical activity, and toxicity of single-agent 5-FU and 5-FU-modulated regimens are reviewed. PMID- 9420017 TI - Perspectives on new chemotherapeutic agents in the treatment of colorectal cancer. AB - In patients with metastatic colorectal cancer (CRC), conventional chemotherapy with 5-fluorouracil (5-FU) plus leucovorin provides an overall response rate of approximately 25% but has had little effect on survival. Thus, alternate agents, new combinations of agents, and new treatment strategies are being investigated. Research efforts over the past decade have increased our understanding of how anticancer agents mediate their antitumor effects, and specific targets for inhibiting the survival, growth, or metastasis of CRC cells have been elucidated. Advances in our understanding have led not only to improvements in the application of currently available agents, but also to the discovery of new agents with activity in CRC. The following active areas of research and/or treatment approaches are discussed: (1) approaches for enhancing 5-FU/leucovorin activity; (2) novel delivery of 5-FU or 5-FU precursor agents; (3) new thymidylate synthase inhibitors; (4) new platinum analogues; (5) topoisomerase I inhibitors; (6) targeting specific proteins or pathways important for the growth, survival, or metastasis of CRC cells; (7) biologic response modifiers, including monoclonal antibodies; and (8) gene therapy. As the cellular mechanisms involved in CRC are further defined and chemotherapy or biologic agents more precisely targeted, response rates and ultimately survival will hopefully improve in this patient population. PMID- 9420018 TI - The role of radiation therapy in rectal cancer. AB - Radiation therapy is an effective but local modality in the treatment of colorectal cancer, with a more clearly defined role in rectal cancer than in colon cancer. In patients with resectable rectal cancer, randomized trials reveal an improvement in local control and survival with postoperative combined modality therapy. Radiation therapy combined with 5-fluorouracil-based chemotherapy appears to be the most effective regimen. Additional benefits with preoperative combined modality therapy may include greater sphincter preservation and less acute toxicity than with postoperative therapy. In patients with locally advanced/unresectable disease, preoperative combined modality therapy increases resectability and may improve local failure rates and overall survival. PMID- 9420019 TI - Folate and antifolate pharmacology. AB - Folic acid is a water-soluble vitamin associated with the other B vitamins. In its fully reduced form (tetrahydrofolate), folate serves as a 1-carbon donor for synthesis of purines and thymidine as well as in the remethylation cycle of homocysteine to methionine. Folate is essential for normal cell growth and replication. It therefore is not surprising that folate analogues have served and continue to serve well as antibiotics and cytotoxic drugs in the treatment of cancer, autoimmune diseases, psoriasis, and bacterial and protozoal infections. During the past 50 years, many of the enzymes requiring folate as a co-factor (ie, thymidylate synthase), and molecules critical in folate homeostasis (ie, the reduced folate carrier, folylpolyglutamate synthase), have been purified and even crystallized. The genes have been cloned, sequenced, and mapped, providing detailed knowledge of their regulation and three-dimensional structure. This has, in part, led to the rational synthesis of a large number of folate analogues that differ from methotrexate, the "classical antifolate," in transport, metabolism, and intracellular targets. Currently, several new folate analogues with unique biochemical properties and clinical applications are being tested. The goals of this brief review are to review folate homeostasis, to highlight the similarities and differences between natural folate and antifolates with respect to biochemistry and metabolism, and to present the pharmacology of methotrexate and several next-generation folate analogues, such as trimetrexate and raltritrexed, with an emphasis on mechanisms of drug resistance. PMID- 9420021 TI - Biomodulation of 5-fluorouracil with antifolates. AB - The cytotoxic activity of 5-fluorouracil (5-FU) can be modulated by coadministration of antifolates or leucovorin (LV). Although a recent meta analysis concluded that a sequential combination of methotrexate (MTX) and 5-FU was superior to 5-FU alone in terms of response rate and survival, combination MTX and 5-FU therapy has not been actively pursued by many leading cancer centers. We have subsequently investigated the combination of trimetrexate (TMTX) plus 5-FU/LV. Unlike MTX, TMTX does not compete with LV for uptake or polyglutamylation. In a phase I clinical study, combination TMTX/5-FU/LV was well tolerated and produced an overall response rate of 20% in previously treated colorectal cancer patients. In a follow-up phase II clinical study, this combination was highly active in patients with advanced colorectal cancer, demonstrating a 50% overall response rate. Currently, a phase III clinical trial is in progress comparing this regimen with combination 5-FU/LV. PMID- 9420020 TI - Antifolates in clinical development. AB - Many novel antifolate compounds with unique pharmacologic properties are currently in clinical development. These newer antifolates differ from methotrexate, the most widely used and studied drug in this class, in terms of their lipid solubility and cellular transport affinity, their level of polyglutamation, and their specificity for inhibiting folate-dependent enzymes, such as dihydrofolate reductase, thymidylate synthase, or glycinamide ribonucleotide formyltransferase. The current status (ie, mechanism of action, clinical response rates, and toxicity) of some of the newer antifolate compounds presently in clinical testing, including edatrexate, piritrexim, raltritrexed, LY 231514, AG337, AG331, 1843U89, ZD 9331, and lometrexol, is reviewed. PMID- 9420022 TI - Trimetrexate: review and current clinical experience in advanced colorectal cancer. AB - Unresectable metastatic colorectal cancer remains a significant cause of morbidity and mortality in both the United States and Europe. To date, no chemotherapeutic regimen for this disease has demonstrated a definitive survival advantage compared with 5-fluorouracil (5-FU) plus leucovorin (LV). However, recent trials have raised the possibility that the combination of trimetrexate (TMTX) plus 5-FU/LV may improve response rates and survival in patients with metastatic colorectal cancer. Trimetrexate is a nonclassical antifolate that has demonstrated antitumor activity against a number of malignancies, including those resistant to the classical antifolate methotrexate. While the single-agent activity of TMTX remains modest in metastatic colorectal cancer, the combination of TMTX/5-FU/LV has shown significant activity in several phase II trials. Reported studies include a phase II trial in chemotherapy failures that demonstrated a 20% response rate, and two multicenter phase II trials in previously untreated patients that demonstrated 50% and 38% overall response rates, respectively. Diarrhea was the dose-limiting toxicity in all trials, although a regimen of scheduled loperamide was quite effective in mitigating this complication. These studies are being followed up with two confirmatory phase II studies in chemorefractory patients and two randomized phase III trials comparing TMTX/5-FU/LV with standard 5-FU/LV. PMID- 9420023 TI - Nursing care strategies for the management of side effects in patients treated for colorectal cancer. AB - Oncology nurses play a critical role in the detection and management of adverse effects resulting from the toxicity of colorectal cancer (CRC) treatment regimens. Standard chemotherapy for CRC involves combination 5-fluorouracil plus leucovorin, a regimen with a well-characterized toxicity profile that includes abdominal cramping and diarrhea, nausea and vomiting, skin and hypersensitivity reactions, fatigue, stomatitis, neutropenia and thrombocytopenia, and alopecia. Diarrhea is the principal dose-limiting toxicity. Trimetrexate, a nonclassical antifolate, is currently being investigated in combination with 5 fluorouracil/leucovorin in phase II/III trials. In addition to the management of side effects, the psychosocial and educational needs of CRC patients require attention. The rigorous treatment schedule presents patients with multiple obstacles in daily living, significantly impacting their quality of life. The oncology nurse is vital in managing the care of CRC patients and ensuring that their physical, psychosocial, and educational needs are met. Educating patients about adverse treatment effects empowers them to manage their symptoms and enables them to alleviate serious or life-threatening treatment complications. Three case studies are provided to illustrate and reinforce nursing management strategies for hypersensitivity reactions, fatigue, and psychosocial issues related to CRC treatment. PMID- 9420024 TI - Trimetrexate: experience with solid tumors. AB - Trimetrexate (TMTX), a potent inhibitor of the enzyme dihydrofolate reductase, is biochemically and metabolically similar to methotrexate (MTX). Fundamental differences between TMTX and MTX, however, mandate investigation of TMTX in both MTX-sensitive and MTX-resistant tumors. In a number of phase II clinical trials, the antitumor activity of single-agent TMTX has been variable, in part because of the heterogeneity of doses and schedules used and in part because of diverse patient populations. Single-agent activity has been documented in some commonly occurring tumors, including breast, non-small cell lung, and head and neck cancers. Other tumors with sensitivity to single-agent TMTX include transitional cell carcinomas of the urothelium, prostate cancer, and gastric carcinoma. In a small series of children with renal cell carcinoma, significant clinical activity was suggested. The single-agent activity of TMTX, coupled with the finding that TMTX may act as a biochemical modulator of 5-fluorouracil/leucovorin, suggests that the addition of TMTX to 5-fluorouracil/leucovorin should be investigated further. The possibility that TMTX may both exhibit single-agent activity and modulate the effectiveness of other agents makes combination therapy attractive. PMID- 9420025 TI - Administration of aminoglycosides to hemodialysis patients immediately before dialysis: a new dosing modality. AB - We describe a new modality for administering aminoglycosides to hemodialysis (HD) patients, namely, a modification of the once-daily regimen which consists of administering the aminoglycosides over 60 min by drip infusion just before each HD session, with a preplanned peak concentration being reached at the beginning of the session and then with a rapidly decreasing concentration being achieved by the start of HD. The area under the concentration-time curve (AUC), i.e., the accumulation of the drug in the body, is thus minimized by this modality. Arbekacin (ABK) was given at a dose of 2 mg/kg of body weight to 10 HD patients infected with methicillin-resistant Staphylococcus aureus (MRSA) for 2 weeks (six sessions in total), resulting in the complete disappearance of MRSA in 5 patients. A high rate of elimination of ABK was attained for each patient while the patient was on HD (range, 0.20 to 0.42 h-1; mean 0.28 +/- 0.08 h-1) by using high-performance dialyzers provided with membranes made of either polymethylmethacrylate, cellulose triacetate (CTA), or ethylene vinyl alcohol. The best results were obtained with the CTA membrane, as revealed by the overall mass transfer coefficient (Ko). The AUC in the simulation model for the variation in the serum ABK concentration in this modality was calculated to be 40% of that of the conventional post-HD dosing modality, suggesting that a much higher dose could be administered to HD patients who receive HD thrice weekly (4 h per session), giving, e.g., 4 mg/kg initially and before the HD sessions, when there is an interval of 68 h from HD session to HD session, and giving 2 mg/kg before the other sessions. PMID- 9420026 TI - Appearance of a metronidazole-resistant Helicobacter pylori strain in an infected ICR-mouse model and difference in eradication of metronidazole-resistant and sensitive strains. AB - We tested whether antibiotic-resistant strains appeared in vivo after the failure of treatment using the Helicobacter pylori-infected euthymic mouse model. The numbers of colonies isolated from 56 ICR mice 2 weeks after 4 days of treatment with metronidazole (3.2, 10, or 32 mg/kg of body weight) or amoxicillin (1, 3.2 or 10 mg/kg), with treatment started 4 days after H. pylori CPY2052 inoculation, were counted, and the isolated strains were tested for their sensitivities to two antibiotics to rule out the presence of antibiotic-resistant strains. One metronidazole-resistant strain was detected in a mouse treated with 10 mg of metronidazole per kg, and the MIC of metronidazole for this strain was 25 microg/ml, compared to a MIC of 1.56 microg/ml for the original strain. However, no resistant strain was detected in the amoxicillin treatment group. After the examination described above, mice challenged with a metronidazole-resistant or sensitive strain isolated from the stomach of a mouse were treated with metronidazole or amoxicillin. The metronidazole-resistant strain was more difficult to eradicate in vivo than the sensitive strain after treatment with metronidazole but not after treatment with amoxicillin. Thus, a metronidazole resistant H. pylori strain was selected by insufficient treatment, but no resistant strain was selected with amoxicillin. Eradication of a metronidazole resistant H. pylori strain in vivo required a higher dosage than eradication of a metronidazole-sensitive H. pylori strain. These results may explain one of the reasons for H. pylori treatment failure. PMID- 9420027 TI - Nosocomial spread of cephem-resistant Escherichia coli strains carrying multiple Toho-1-like beta-lactamase genes. AB - Escherichia coli HKY56, which demonstrated resistance to various beta-lactams except carbapenems, was isolated from the throat swab of an inpatient in 1994. Conjugal transfer of cephem resistance from HKY56 to E. coli CSH2 was not successful. Three cefotaxime-resistant E. coli clones harboring plasmid pMRE001, pMRE002, or pMRE003, each of which carried a 3.4-, 5.8-, or 6.2-kb EcoRI fragment insert, respectively, were obtained from HKY56. Although restriction analysis suggested their different origins, these clones showed similar profiles of resistance to various beta-lactams. The sequence of 10 amino acid residues at the N terminus of beta-lactamase purified from E. coli HB101(pMRE001) was identical to that of Toho-1. This Toho-1-like beta-lactamase-1 (TLB-1) was able to hydrolyze cefoperazone and cefotaxime efficiently, but it failed to hydrolyze cephamycins. A Toho-1-specific DNA probe was hybridized with three distinct EcoRI fragments derived from the chromosomal DNA of strain HKY56, and these fragments corresponded to DNA inserts carried by pMRE001, pMRE002, and pMRE003, respectively. PCR and Southern hybridization analysis suggested that all six cephem-resistant E. coli strains, strains HKY273, HKY285, HKY288, HKY305, HKY316, and HKY335, which were isolated in 1996 at the same hospital where strain HKY56 had been isolated, also possessed multiple Toho-1-like beta-lactamase (TLB) genes, and the hybridization patterns obtained with the Toho-1-specific probe were quite similar among these six isolates. The DNA fingerprinting patterns observed by pulsed-field gel electrophoresis revealed that among the E. coli isolates tested, all isolates except HKY56 possessed a similar genetic background. These findings suggested that E. coli strains that carry chromosomally multiplied TLB genes may have been proliferating and transmitted among patients in the same hospital. PMID- 9420028 TI - In vitro activities of beta-lactam-beta-lactamase inhibitor combinations against Stenotrophomonas maltophilia: correlation between methods for testing inhibitory activity, time-kill curves, and bactericidal activity. AB - The activities of ampicillin, ampicillin-sulbactam, amoxicillin, amoxicillin clavulanic acid, ticarcillin, ticarcillin-clavulanic acid, piperacillin, piperacillin-tazobactam, aztreonam, and aztreonam-clavulanic against Stenotrophomonas maltophilia strains for which the MICs of penicillins and commercially available beta-lactam-beta-lactamase inhibitor combinations were higher than the breakpoints usually recommended for Pseudomonas aeruginosa in commercially available broth microdilution methods were tested by the agar diffusion, agar dilution, and broth microdilution methods. Time-kill curve studies were performed when discrepancies between these methods were observed. The MICs obtained by the commercially available broth microdilution method, the agar dilution method, and the broth microdilution method were almost identical. Twenty-five percent of the strains tested showed inhibition diameters of > or =15 mm for ticarcillin-clavulanic acid, and 43.7% of the strains tested showed inhibition diameters of > or =18 mm for piperacillin-tazobactam by the agar diffusion method. The time-kill curves for these strains confirmed the results obtained by dilution methods. Aztreonam-clavulanic acid (2:1) at concentrations of < or =16 microg/ml inhibited all of these strains (MIC range, 1 to 16 microg/ml). The time-kill curves confirmed this activity. The addition of piperacillin to this combination did not modify the MICs. The combination aztreonam-clavulanic acid-ticarcillin was two- to fourfold more active than aztreonam-clavulanic acid alone. We studied the inhibitory and bactericidal activities of the two most active combinations (aztreonam-clavulanic acid and aztreonam-clavulanic acid-ticarcillin) against the standard inoculum and 10 and 50 times the standard inoculum. Inoculum modifications did not modify the MICs. Both combinations showed good bactericidal activity against the standard inoculum. With 10 times the standard inoculum, minimum bactericidal concentration (MBC) results were heterogeneous (for 55% of the strains, MBCs were between the MIC and 4-fold the MIC, and for 45% of the strains MBCs were between 8- and >32 fold the MIC). With 50 times the standard inoculum, MBCs were at least 32-fold the MICs for all the strains tested. PMID- 9420029 TI - Sensitivity of human immunodeficiency virus to bicyclam derivatives is influenced by the three-dimensional structure of gp120. AB - The bicyclams are a new class of anti-human immunodeficiency virus (anti-HIV) compounds targeted at viral entry. From marker rescue experiments, it appears that the envelope gp120 glycoprotein plays an important role in the anti-HIV activity of the bicyclams. Bicyclam-resistant strains contain a number of amino acid changes scattered over the V2 to V5 region of gp120. Experiments aimed at estimating the relative importance of particular amino acid changes with regard to the overall resistance pattern are described. The sequences of some partially bicyclam-resistant virus strains, obtained during the resistance development process, were analyzed, and the corresponding 50% effective concentrations were determined. Selected mutations observed in bicyclam-resistant strains were introduced in the wild-type background by site-directed mutagenesis. In addition, some amino acids were back-mutated to their wild-type counterparts in an otherwise JM3100-resistant strain. The sensitivities of these mutant viruses to bicyclams were determined. Construction of chimeric viruses, carrying the V3 loop of JM3100-resistant virus in a wild-type HIV type 1 HXB2 background, enabled us to investigate the importance of the mutations in the V3 loop of JM3100-resistant virus. From the results described in the report, it can be concluded that single amino acid substitutions do not influence the observed resistance to JM3100. Also, the mutations in the V3 loop are not sufficient to engender even a partially resistant phenotype. We postulate that the overall conformation of gp120 determines the degree of sensitivity or resistance of HIV strains to bicyclams. PMID- 9420030 TI - Cloning and sequence analysis of two copies of a 23S rRNA gene from Helicobacter pylori and association of clarithromycin resistance with 23S rRNA mutations. AB - In this study, two identical copies of a 23S-5S gene cluster, which are separately situated within the Helicobacter pylori UA802 chromosome, were cloned and sequenced. Comparison of the DNA sequence of the H. pylori 23S rRNA gene with known sequences of other bacterial 23S rRNA genes indicated that the H. pylori UA802 23S rRNA genes are closely related to those of Campylobacter spp. and therefore belong in the proposed Proteobacteria subdivision. The 5'-terminal nucleotide T or A of the 23S rRNA is close to a Pribnow box which could be a -10 region of the transcription promoter for the 23S rRNA gene, suggesting that a posttranscriptional process is likely not involved in the maturation of the H. pylori 23S rRNA. Clinical isolates of H. pylori resistant to clarithromycin were examined by using natural transformation and pulsed-field gel electrophoresis. Cross-resistance to clarithromycin and erythromycin, which was transferred by natural transformation from the Cla(r) Ery(r) donor strain H. pylori E to the Cla(s) Ery(s) recipient strain H. pylori UA802, was associated with an single A to-G transition mutation at position 2142 of both copies of the 23S rRNA in UA802 Cla(r) Ery(r) mutants. The transformation frequency for Cla(r) and Ery(r) was found to be approximately 2 x 10(-6) transformants per viable cell, and the MICs of both clarithromycin and erythromycin for the Cla(r) Ery(r) mutants were equal to those for the donor isolate. Our results confirmed the previous findings that mutations at positions 2142 and 2143 of the H. pylori 23S rRNA gene are responsible for clarithromycin resistance and suggest that acquisition of clarithromycin resistance in H. pylori could also result from horizontal transfer. PMID- 9420031 TI - A single point mutation in the embB gene is responsible for resistance to ethambutol in Mycobacterium smegmatis. AB - Ethambutol [EMB; dextro-2,2'-(ethylenediimino)-di-1-butanol] is an effective drug when used in combination with isoniazid for the treatment of tuberculosis. It inhibits the polymerization of arabinan in the arabinogalactan and lipoarabinomannan of the mycobacterial cell wall. Recent studies have shown that arabinosyltransferases could be targets of EMB. These enzymes are encoded by the emb locus that was identified in Mycobacterium smegmatis, Mycobacterium leprae, Mycobacterium avium, and Mycobacterium tuberculosis. We demonstrate that a missense mutation in the M. smegmatis embB gene, one of the genes of the emb locus, confers resistance to EMB. The level of resistance is not dependent on the number of copies of the mutated embB gene, indicating that this is a true mechanism of resistance. The mutation is located in a region of the EmbB protein that is highly conserved among the different mycobacterial species. We also identified in this region two other independent mutations that confer EMB resistance. Furthermore, mutations have recently been described in the same region of the EmbB protein from clinical EMB-resistant M. tuberculosis isolates. Together, these data strongly suggest that one of the mechanisms of resistance to EMB consists of missense mutations in a particular region of the EmbB protein that could be directly involved in the interaction with the EMB molecule. PMID- 9420032 TI - Antimicrobial susceptibility testing of 230 Helicobacter pylori strains: importance of medium, inoculum, and incubation time. AB - No standardized method of susceptibility testing for Helicobacter pylori is currently available, so before a large agar dilution study comprising 230 H. pylori strains belonging to more than 80 genetically different groups was initiated, we performed a relatively small preliminary study to determine the influences of medium, inoculum density, and incubation time. Seven media were investigated and were primarily evaluated on the basis of their abilities to support growth both semiquantitatively and qualitatively; Iso-Sensitest agar supplemented with 10% horse blood was found to be well suited for the purpose; this was closely followed by Mueller-Hinton agar with 10% horse blood, Mueller Hinton with 10% sheep blood, and finally, 7% lysed horse blood agar. Investigations of two inoculum densities and two incubation times resulted in recommendations for the use of 10(9) CFU/ml (10[6] CFU/spot) as the inoculum and 72 h as the incubation time. A modest inoculum effect was noted for amoxicillin and metronidazole. By the methodology derived from our preliminary study, the susceptibilities of 230 H. pylori strains to six antibiotics were subsequently determined. The results were generally in accord with those of others, and apart from metronidazole, the MIC of which for approximately 25% of the strains tested was >8 microg/ml, resistance was low in Denmark. The situation might, however, quickly change when and if the number of indications for antibiotic therapy for H. pylori infections increase. Consequently, susceptibility testing of all H. pylori strains is recommended in order to survey the development of resistance, and in our hands the described methodology was relatively easy to perform and the results were easy to read. PMID- 9420033 TI - Pharmacokinetics of imipenem-cilastatin in critically ill patients undergoing continuous venovenous hemofiltration. AB - The pharmacokinetics of imipenem-cilastatin were investigated in 12 critically ill patients with acute renal failure (ARF) managed by continuous veno-venous hemofiltration (CVVH) while receiving a fixed combination of 500 mg of imipenem cilastatin intravenously three or four times daily. No adverse drug reactions were observed. Plasma and hemofiltrate samples were taken at specified times during one dosing interval, and the concentrations of imipenem and cilastatin were determined by high-performance liquid chromatography. Pharmacokinetic variables were calculated by a first-order, two-compartment pharmacokinetic model for both substances. Total clearances of imipenem and cilastatin (mean +/- standard deviations) were 122.2 +/- 28.6 and 29.2 +/- 13.7 ml/min, respectively, with hemofiltration clearances of 22.9 +/- 2.5 and 16.1 +/- 3.1 ml/min, respectively, and nonrenal, nonhemofiltration clearances of 90.8 +/- 26.3 and 13.2 +/- 13.9 ml/min, respectively. Mean imipenem dosage requirements were approximately 2,000 mg/24 h (2,111.8 +/- 493.4 mg/24 h). They were calculated in order to achieve an average steady-state concentration of 12 mg/liter to ensure that concentrations in plasma exceeded the MICs at which 90% of intermediately resistent bacteria are inhibited (8 mg/liter) during the majority of the dosing interval. By contrast, the recommended dosage for patients with end-stage renal failure (ESRF) and infections caused by intermediately resistant bacteria is 1,000 mg/24 h. This remarkable difference may be due (i) to differences in the nonrenal clearance of imipenem between patients with ARF and ESRF and (ii) to the additional clearance by the hemofilter. Since the total clearance of cilastatin was low, marked accumulation occurred, and this was particularly pronounced in patients with additional liver dysfunction. Thus, in patients with ARF managed by CVVH, rather high imipenem doses are required, and these inevitably result in a marked accumulation of cilastatin. The doses of imipenem recommended for patients with ESRF, however, will lead to underdosing and inadequate antibiotic therapy. PMID- 9420034 TI - Loss of intrinsic aminoglycoside resistance in Acinetobacter haemolyticus as a result of three distinct types of alterations in the aac(6')-Ig gene, including insertion of IS17. AB - The distribution of the aac(6')-Ig gene, encoding aminoglycoside 6'-N acetyltransferase-Ig [AAC(6')-Ig], was studied in 96 Acinetobacter haemolyticus strains and 12 proteolytic Acinetobacter strains, including Acinetobacter genomospecies 6, 13, and 14 and 3 unnamed species assigned to this genomic group by DNA-DNA hybridization. This gene was detected by DNA-DNA hybridization in all 96 A. haemolyticus strains and by PCR in 95 strains but was not detected in strains of other species, indicating that it may be used to identify A. haemolyticus. Three A. haemolyticus strains were susceptible to tobramycin and did not produce an aminoglycoside 6'-N-acetylating activity, although they contained aac(6')-Ig-related sequences. An analysis of three susceptible A. haemolyticus strains indicated that aminoglycoside resistance was abolished by the following three distinct mechanisms: (i) a point mutation in aac(6')-Ig that led to a Met56-->Arg substitution, which was shown by analysis of a revertant to be responsible for the loss of resistance; (ii) a polythymine insertion that altered the reading frame; and (iii) insertion of IS17, a new member of the IS903 family. These observations indicated that AAC(6')-Ig is not essential for the viability of A. haemolyticus, although the aac(6')-Ig gene was detected in all members of this species. PMID- 9420035 TI - In vitro and in vivo antibacterial activities of CS-834, a novel oral carbapenem. AB - CS-834 is a novel oral carbapenem antibiotic. This compound is an ester-type prodrug of the active metabolite R-95867. The antibacterial activity of R-95867 was tested against 1,323 clinical isolates of 35 species and was compared with those of oral cephems, i.e., cefteram, cefpodoxime, cefdinir, and cefditoren, and that of a parenteral carbapenem, imipenem. R-95867 exhibited a broad spectrum of activity covering both gram-positive and -negative aerobes and anaerobes. Its activity was superior to those of the other compounds tested against most of the bacterial species tested. R-95867 showed potent antibacterial activity against clinically significant pathogens: methicillin-susceptible Staphylococcus aureus including ofloxacin-resistant strains, Streptococcus pneumoniae including penicillin-resistant strains, Clostridium perfringens, Neisseria spp., Moraxella catarrhalis, most members of the family Enterobacteriaceae, and Haemophilus influenzae (MIC at which 90% of strains are inhibited, < or =0.006 to 0.78 microg/ml). R-95867 was quite stable to hydrolysis by most of the beta-lactamases tested except the metallo-beta-lactamases from Stenotrophomonas maltophilia and Bacteroides fragilis. R-95867 showed potent bactericidal activity against S. aureus and Escherichia coli. Penicillin-binding proteins 1 and 4 of S. aureus and 1Bs, 2, 3, and 4 of E. coli had high affinities for R-95867. The in vivo efficacy of CS-834 was evaluated in murine systemic infections caused by 16 strains of gram-positive and -negative pathogens. The efficacy of CS-834 was in many cases superior to those of cefteram pivoxil, cefpodoxime proxetil, cefdinir, and cefditoren pivoxil, especially against infections caused by S. aureus, penicillin resistant S. pneumoniae, E. coli, Citrobacter freundii, and Proteus vulgaris. Among the drugs tested, CS-834 showed the highest efficacy against experimental pneumonia in mice caused by penicillin-resistant S. pneumoniae. PMID- 9420036 TI - Safety and pharmacokinetics of CS-834, a new oral carbapenem antibiotic, in healthy volunteers. AB - CS-834, (+)-[pivaloyloxymethyl (4R,5S,6S)-6-[(R)-1-hydroxyethyl]-4-methyl-7-oxo-3 [[(R)-5-oxopyrroli din-3-yl]thio]-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate], is an ester-type oral carbapenem prodrug, and an active metabolite is R-95867, which has antibacterial activity. CS-834 was administered orally to healthy male volunteers at single doses of 50, 100, 200, and 400 mg and at a multiple dose of 150 mg three times a day for 7 days to investigate its safety and pharmacokinetic profiles. Other studies were conducted to examine the effect of food intake on the bioavailability of CS-834 and also the effect of the coadministration of probenecid on the pharmacokinetics of CS-834. In the fasting state, the concentration of R-95867 in plasma reached maximum levels from 1.1 to 1.7 h after the oral administration of CS-834, followed by a monoexponential decrease. The maximum concentrations of R-95867 in serum (C[max]s) after the administration of CS-834 at doses of 50, 100, 200, and 400 mg were 0.51, 0.97, 1.59, and 2.51 microg/ml, respectively. The half-lives (t1/2s) were almost constant, approximately 0.7 h. The areas under the concentration-time curves (AUCs) were proportional to the doses, ranging from 50 to 400 mg x h/ml. The cumulative recoveries in urine were approximately 30 to 35% until 24 h after drug administration. The C(max), AUC, t1/2, and recovery in urine were not affected by food intake. Probenecid coadministration prolonged the t1/2, and it increased the C(max) and AUC for R-95867 by approximately 1.5- and 2.1-fold, respectively. The multiple-dose study showed no change in the pharmacokinetics from those for the single doses and no drug accumulation in the body. A mild transient soft stool was observed in one volunteer in the study with a single dose of 400 mg. In the multiple-dose study, mild transient soft stools were observed in six volunteers, one volunteer had mild transient diarrhea, and one volunteer had elevated serum glutamic oxalacetic transaminase and serum glutamic pyruvic transaminase levels (1.4- and 2.8-fold compared with the upper limits of normal, respectively). There were no other abnormal findings for objective symptoms or laboratory findings, including blood pressure, heart rate, electrocardiogram, body temperature, hematology, blood chemistry, and urinalysis. PMID- 9420037 TI - Population pharmacokinetic modeling of isoniazid, rifampin, and pyrazinamide. AB - Isoniazid (INH), rifampin (RIF), and pyrazinamide (PZA) are the most important drugs for the treatment of tuberculosis (TB). The pharmacokinetics of all three drugs in the plasma of 24 healthy males were studied as part of a randomized cross-over phase I study of two dosage forms. Subjects ingested single doses of INH at 250 mg, RIF at 600 mg, and PZA at 1,500 mg. Plasma was collected for 36 h and was assayed by high-performance liquid chromatography. The data were analyzed by noncompartmental, iterative two-stage maximum a posteriori probability Bayesian (IT2B) and nonparametric expectation maximization (NPEM) population modeling methods. Fast and slow acetylators of INH had median peak concentrations in plasma (C[max]) of 2.44 and 3.64 microg/ml, respectively, both of which occurred at 1.0 h postdose (time of maximum concentrations of drugs in plasma [T(max)]), with median elimination half-lives (t1/2) of 1.2 and 3.3 h, respectively (by the NPEM method). RIF produced a median C(max) of 11.80 microg/ml, a T(max) of 1.0 h, and a t1/2 of 3.4 h. PZA produced a median C(max) of 28.80 microg/ml, a T(max) of 1.0 h, and a t1/2 of 10.0 h. The pharmacokinetic behaviors of INH, RIF, and PZA were well described by the three methods used. These models can serve as benchmarks for comparison with models for other populations, such as patients with TB or TB with AIDS. PMID- 9420038 TI - Lobucavir is phosphorylated in human cytomegalovirus-infected and -uninfected cells and inhibits the viral DNA polymerase. AB - Lobucavir (LBV) is a deoxyguanine nucleoside analog with broad-spectrum antiviral activity. LBV was previously shown to inhibit herpes simplex virus (HSV) DNA polymerase after phosphorylation by the HSV thymidine kinase. Here we determined the mechanism of action of LBV against human cytomegalovirus (HCMV). LBV inhibited HCMV DNA synthesis to a degree comparable to that of ganciclovir (GCV), a drug known to target the viral DNA polymerase. The expression of late proteins and RNA, dependent on viral DNA synthesis, was also inhibited by LBV. Immediate early and early HCMV gene expression was unaffected, suggesting that LBV acts temporally coincident with HCMV DNA synthesis and not through cytotoxicity. In vitro, the triphosphate of LBV was a potent inhibitor of HCMV DNA polymerase with a Ki of 5 nM. LBV was phosphorylated to its triphosphate form intracellularly in both infected and uninfected cells, with phosphorylated metabolite levels two- to threefold higher in infected cells. GCV-resistant HCMV isolates, with deficient GCV phosphorylation due to mutations in the UL97 protein kinase, remained sensitive to LBV. Overall, these results suggest that LBV-triphosphate halts HCMV DNA replication by inhibiting the viral DNA polymerase and that LBV phosphorylation can occur in the absence of viral factors including the UL97 protein kinase. Furthermore, LBV may be effective in the treatment of GCV resistant HCMV. PMID- 9420039 TI - Flow cytometric analysis of herpes simplex virus type 1 susceptibility to acyclovir, ganciclovir, and foscarnet. AB - We established a quantitative flow cytometric method for determination of herpes simplex virus type 1 (HSV-1) susceptibility to acyclovir (ACV), ganciclovir, and foscarnet in vitro. Susceptibility was defined in terms of the drug concentration which reduced the number of cells expressing HSV-1 glycoprotein C (gpC) with a fluorescence intensity of > or =10(2) by 50% (IC50). Flow cytometry allowed us to use a high (1.0) as well as a low (0.005) multiplicity of infection, and determination of the IC50 was possible after one or more viral replicative cycles. IC50s were dependent on virus input and on time postinfection. In mixture experiments, 1 to 2% resistant viruses added to a sensitive strain could be detected. The results obtained by flow cytometry showed a good qualitative correlation with those achieved by cytopathic effect inhibitory assay. However, flow cytometry might detect more quantitative differences in drug susceptibility, especially among resistant strains, as confirmed also by determination of intracellular drug phosphorylation. The mean IC50s for ACV-sensitive strains were 0.45 to 1.47 microM, and those for ACV-resistant strains were between 140 and 3,134 microM. Flow cytometric analysis was fast and accurate, automatizable, and highly reproducible. Flow cytometry may be a more powerful tool than standard cytopathic effect-based assays and could have advantages for the detection of low levels of drug resistance or mixtures of sensitive and resistant virus strains. PMID- 9420040 TI - Transepithelial transport of the fluoroquinolone ciprofloxacin by human airway epithelial Calu-3 cells. AB - Although fluoroquinolone antibiotics such as ciprofloxacin are able to gain access to lung tissue and both pleural and bronchial secretions, the characteristics of transport and cellular uptake of ciprofloxacin in human epithelial lung tissue remain obscure. We have chosen human airway epithelial (Calu-3) cells, reconstituted as functional epithelial layers grown on permeable filter supports, as a model with which to assess both transepithelial transport and cellular uptake of ciprofloxacin. Transepithelial ciprofloxacin fluxes in absorptive (apical-to-basal) and secretory (basal-to-apical) directions were similar throughout the concentration range studied (1.0 microM to 3.0 mM). Transepithelial mannitol fluxes measured concurrently were substantially smaller than ciprofloxacin fluxes in Calu-3 epithelia, suggesting the existence of a mediated transcellular route in addition to a paracellular route for transepithelial permeation. Apical-to-basal ciprofloxacin flux (at 10 microM) was inhibited by a 100-fold excess of unlabelled norfloxacin, enoxacin, and ofloxacin, while secretory flux was unaffected. Cellular uptake of ciprofloxacin, determined as a cell/medium ratio, was greater from the basal compartment (2.7 fold) than apical uptake (1.39-fold) measured at 100 microM ciprofloxacin and showed no saturation up to 3 mM ciprofloxacin. Comparison of the permeation of ciprofloxacin was made with that of lipophilic substrates such as vinblastine and digoxin. There was a linear correlation between transepithelial permeability (Pa b) and their oil/water partition coefficients with mannitol < ciprofloxacin < digoxin < vinblastine. Comparison of transport of ciprofloxacin across human airway Calu-3 epithelia with that across intestinal Caco-2 epithelia emphasizes the absence of a specific secretory pathway; ciprofloxacin permeation in Calu-3 epithelia appears to be mediated primarily by a transcellular route, with mediated transfer at apical and basal membranes occurring via transporters with low affinity to ciprofloxacin. PMID- 9420041 TI - Inhibition of the multiple antibiotic resistance (mar) operon in Escherichia coli by antisense DNA analogs. AB - The multiple antibiotic resistance operon (marORAB) in Escherichia coli controls intrinsic susceptibility and resistance to multiple, structurally different antibiotics and other noxious agents. A plasmid construct with marA cloned in the antisense direction reduced LacZ expression from a constitutively expressed marA::lacZ translational fusion and inhibited the induced expression of LacZ in cells bearing the wild-type repressed fusion. The marA antisense construction also decreased the multiple antibiotic resistance of a Mar mutant. Two antisense phosphorothioate oligonucleotides, one targeted to marO and the other targeted to marA of the mar operon, introduced by heat shock or electroporation reduced LacZ expression in the strain having the marA::lacZ fusion. One antisense oligonucleotide, tested against a Mar mutant of E. coli ML308-225, increased the bactericidal activity of norfloxacin. These studies demonstrate the efficacy of exogenously delivered antisense oligonucleotides targeted to the marRAB operon in inhibiting expression of this chromosomal regulatory locus. PMID- 9420043 TI - Fungicidal mechanism of action of D0870 against Cryptococcus neoformans under acidic conditions. AB - The fungicidal mechanism of the triazole D0870 against Cryptococcus neoformans under acidic conditions was investigated. D0870 reduced the intracellular K+ content of C. neoformans at pH 4 to about half the value at pH 7 after 12 h of incubation. The 50% inhibitory concentrations of D0870 for ergosterol biosynthesis were almost the same at both pH 4 (0.017 microg/ml) and 7 (0.014 microg/ml); however, D0870 caused a marked accumulation of an unknown lipid and methylated sterols in C. neoformans cultured at pH 4. Extracted fractions containing the unknown lipid or methylated sterols showed strong fungicidal activities against C. neoformans both at pH 4 and 7 in phosphate-citrate buffer not containing D0870. Gas chromatographic-mass spectrometric analysis showed that the unknown lipid was obtusifolione. These results suggest that D0870 kills C. neoformans by disturbing the permeability of the cell membrane through the accumulation of obtusifolione and methylated sterols in the cell membrane under acidic conditions. PMID- 9420042 TI - An allelic variant of the chromosomal gene for class A beta-lactamase K2, specific for Klebsiella pneumoniae, is the ancestor of SHV-1. AB - Fecal Klebsiella isolates from neonates in 22 Swedish special care units were examined by a PCR we developed for detection of the SHV-1 beta-lactamase gene. All 105 K. pneumoniae isolates and all 11 K. pneumoniae reference strains (including the K. pneumoniae subsp. pneumoniae, ozaenae, and rhinoscleromatis type strains) tested were positive, whereas all 67 K. oxytoca isolates and the K. oxytoca, K. planticola, and K. terrigena type strains tested were negative. Resistance to beta-lactams in K. pneumoniae was not transferable by conjugation, and the beta-lactamase gene was never found on a plasmid. Southern blot analysis showed that the gene had a defined chromosomal location. Isoelectric focusing and sequencing of 231-bp PCR amplicons from different isolates revealed many variants of the enzyme, with the two main groups being SHV-1 like (pI 7.6; 68 isolates) and LEN-1 like (pI 7.1; 14 isolates). Clavulanic acid markedly reduced the MICs of ampicillin for all the K. pneumoniae isolates tested. This fact, MIC profiles (penicillin rather than cephalosporin resistance), pIs, and sequence data showed that the chromosomal beta-lactamase of K. pneumoniae is a class A, group 2 enzyme distinct from the chromosomal AmpC enzymes found in several other gram-negative bacteria and from the chromosomal beta-lactamase K1 of K. oxytoca. We propose that the chromosomal beta-lactamase of K. pneumoniae be designated K2 and suggest that an allelic pI 7.6 variant of this enzyme is the ancestor of the SHV family of plasmid-mediated beta-lactamases. PMID- 9420044 TI - Concentrations in plasma and safety of 7 days of intravenous itraconazole followed by 2 weeks of oral itraconazole solution in patients in intensive care units. AB - Pharmacokinetics and safety of a hydroxy-beta-propyl solution of itraconazole were assessed in 16 patients in an intensive care unit. On the first 2 days, four 1-h infusions of 200 mg were given at 0, 8, 24, and 32 h. From day 3 to 7, inclusive, a single 1-h infusion of 200 mg of itraconazole was given daily. The intravenous (i.v.) treatment was directly followed by repeated administrations of an oral solution of itraconazole at a dosage of either 200 mg once daily or 200 mg twice daily (b.i.d.). During i.v. treatment, steady-state concentrations of itraconazole and hydroxy-itraconazole in plasma were reached within 48 and 96 h, respectively. At the end of i.v. treatment, mean (+/- standard deviation) itraconazole and hydroxy-itraconazole trough concentrations in plasma were 0.344 +/- 0.140 and 0.605 +/- 0.205 microg/ml, respectively. After the 2-week oral follow-up of 200 mg once daily the mean trough concentration had decreased to 0.245 microg/ml, whereas after 200 mg b.i.d. it increased to 0.369 microg/ml. Diarrhea during oral treatment appeared to be dose related and may be due to the solvent hydroxypropyl-beta-cyclodextrin. More severe laboratory abnormalities were noted during the i.v. than the oral treatment phase, probably related to more severe illness in that period of intensive care, but none proved clinically important. These results suggest that plasma itraconazole levels above 0.250 microg/ml may be achieved and maintained with the 1-week i.v. schedule followed by b.i.d. oral administration, whereas the once-daily oral follow-up seems to be a suboptimal treatment. PMID- 9420045 TI - Cloning and characterization of a novel macrolide efflux gene, mreA, from Streptococcus agalactiae. AB - A strain of Streptococcus agalactiae displayed resistance to 14-, 15-, and 16 membered macrolides. In PCR assays, total genomic DNA from this strain contained neither erm nor mef genes. EcoRI-digested genomic DNA from this strain was cloned into lambda Zap II to construct a library of S. agalactiae genomic DNA. A clone, pAES63, expressing resistance to erythromycin, azithromycin, and spiramycin in Escherichia coli was recovered. Deletion derivatives of pAES63 which defined a functional region on this clone that encoded resistance to 14- and 15-membered, but not 16-membered, macrolides were produced. Studies that determined the levels of incorporation of radiolabelled erythromycin into E. coli were consistent with the presence of a macrolide efflux determinant. This putative efflux determinant was distinct from the recently described Mef pump in Streptococcus pyogenes and Streptococcus pneumoniae and from the multicomponent MsrA pump in Staphylococcus aureus and coagulase-negative staphylococci. Its gene has been designated mreA (for macrolide resistance efflux). PMID- 9420046 TI - Macrolide resistance in Helicobacter pylori: rapid detection of point mutations and assays of macrolide binding to ribosomes. AB - Resistance of Helicobacter pylori to macrolides is a major cause of failure of eradication therapies. Single base substitutions in the H. pylori 23S rRNA genes have been associated with macrolide resistance in the United States. Our goal was to extend this work to European strains, to determine the consequence of this mutation on erythromycin binding to H. pylori ribosomes, and to find a quick method to detect the mutation. Seven pairs of H. pylori strains were used, the parent strain being naturally susceptible to macrolides and the second strain having acquired an in vivo resistance during a treatment regimen that included clarithromycin. The identity of the strains was confirmed by random amplified polymorphic DNA testing with two different primers, indicating that resistance was the result of the selection of variants of the infecting strain. All resistant strains were found to have point mutations at position 2143 (three cases) or 2144 (four cases) but never on the opposite DNA fragment of domain V of the 23S rRNA gene. The mutation was A-->G in all cases except one (A-->C) at position 2143. Using BsaI and BbsI restriction enzymes on the amplified products, we confirmed the mutations of A-->G at positions 2144 and 2143, respectively. Macrolide binding was tested on purified ribosomes isolated from four pairs of strains with [14C]erythromycin. Erythromycin binding increased in a dose dependent manner for the susceptible strain but not for the resistant one. In conclusion we suggest that the limited disruption of the peptidyltransferase loop conformation, caused by a point mutation, reduces drug binding and consequently confers resistance to macrolides. Finally, the macrolide resistance could be detected without sequencing by performing restriction fragment length polymorphism with appropriate restriction enzymes. PMID- 9420047 TI - In vitro susceptibilities of the AIDS-associated microsporidian Encephalitozoon intestinalis to albendazole, its sulfoxide metabolite, and 12 additional benzimidazole derivatives. AB - Recent reports have described the successful treatment of Encephalitozoon intestinalis infection in AIDS patients with albendazole. However, this compound is rapidly metabolized in vivo to albendazole sulfoxide, and furthermore it is only 1 of about 15 commercially developed benzimidazole derivatives. To compare the activities of albendazole, albendazole sulfoxide, and other benzimidazoles, an in vitro system involving infection of green monkey kidney cell (E6) monolayers with E. intestinalis spores was developed. After 14 days, the effects of benzimidazoles on spore production were determined. Ten of fourteen derivatives tested, including albendazole, were inhibitory at concentrations of 1 to 10 ng/ml. Derivatives modified at the 1 or 2 position were less active. Albendazole sulfoxide was 1.7-fold more inhibitory than albendazole but significantly less toxic to E6 cells, a finding that explains the clinical efficacy of this compound. Potential alternatives to albendazole are discussed. No albendazole-resistant E. intestinalis mutants were obtained following in vitro selection. PMID- 9420049 TI - Reappraisal of the antimicrobial susceptibilities of Chryseobacterium and Flavobacterium species and methods for reliable susceptibility testing. AB - Several Flavobacterium species, comprising a heterogeneous group of gram-negative bacilli that are capable of causing opportunistic infections in humans, have recently been reclassified as Chryseobacterium or Myroides species. Intrinsically resistant to a number of antibiotics, these organisms have been reported to be susceptible to vancomycin and certain other drugs that are normally active against gram-positive bacteria. By using the National Committee for Clinical Laboratory Standards (NCCLS) broth microdilution procedure, 58 clinical isolates of former flavobacteria (36 Chryseobacterium meningosepticum isolates, 11 C. indologenes isolates, 3 C. gleum isolates, 4 unspeciated former members of Flavobacterium group IIb, and 4 Myroides odoratum isolates) were tested with 23 antibiotics, including conventional and investigational agents. In addition, the broth microdilution results were compared to those generated by agar dilution, E test, and disk diffusion for vancomycin and piperacillin-tazobactam. Compared to the NCCLS microdilution results, there were 7.1 and 17.9% very major errors with piperacillin-tazobactam by agar dilution and E-test, respectively. In addition, there were from 12.1 to 48.3% minor errors with both procedures with vancomycin and piperacillin-tazobactam. The very major and minor error rates were unacceptably high with disk testing of piperacillin-tazobactam; the use of enterococcal vancomycin disk breakpoints (zone diameter of > or =17 mm = susceptible) resulted in >20% minor errors but only one very major error. All of the isolates were susceptible to minocycline; over 90% were susceptible to sparfloxacin, levofloxacin, and clinafloxacin; and 88% were susceptible to rifampin. None was susceptible to vancomycin. When Chryseobacterium or Myroides species are isolated from serious infections, susceptibility testing by broth microdilution should be performed and therapy should be guided by those results. PMID- 9420048 TI - Mechanisms of fluoroquinolone resistance in genetically related strains of Staphylococcus aureus. AB - Fluoroquinolone resistance in Staphylococcus aureus results from amino acid substitutions at particular locations in the DNA gyrase A and B subunits as well as in the topoisomerase IV A subunit and from NorA-mediated efflux. More than one resistance mechanism may be present in a single strain. Fluoroquinolone-resistant derivatives of SA-1199, a methicillin-susceptible S. aureus strain, were selected in vivo or in vitro, and their mechanisms of fluoroquinolone resistance were identified. We found that many of the resistance mechanisms described above can develop in derivatives of a single parent strain, either singly or in combination, and can arise in a single step. Variances in MICs for strains with the same apparent resistance mechanisms likely are due to the presence of new or undetected but established means of fluoroquinolone resistance. NorA-mediated resistance can occur in the apparent absence of topoisomerase mutations and in some strains may be the result of a promoter region mutation causing increased expression of norA. However, increased expression of norA can occur independently of this mutation, suggesting that a regulatory locus for this gene exists elsewhere on the chromosome. PMID- 9420050 TI - In vitro and in vivo antibacterial activities of GV129606, a new broad-spectrum trinem. AB - GV129606 is a new parenteral trinem antibiotic belonging to the beta-lactam class. It combines broad-spectrum activity (against gram-negative and -positive bacteria, aerobes and anaerobes), with high potency and resistance to beta lactamases. Comparative in vitro and in vivo antibacterial activities were determined for GV129606 against more than 400 recent clinical isolates (aerobes, including beta-lactamase producers, and anaerobes), using representative antibacterial agents (meropenem, piperacillin, ceftazidime, cefpirome, ciprofloxacin, and gentamicin for aerobes and metronidazole, cefoxitin, piperacillin, and clindamycin for anaerobes). Against methicillin-susceptible staphylococci and streptococci, GV129606 and meropenem were the most active of the drugs tested. GV129606 showed an MIC for 90% of strains tested (MIC90) ranging from < or =0.015 to 0.06 microg/ml against methicillin-susceptible staphylococci and Streptococcus sanguis, Streptococcus pyogenes, and Streptococcus agalactiae. Against penicillin-susceptible and -resistant Streptococcus pneumoniae isolates, GV129606, meropenem, and cefpirome showed MIC90s of < or =0.015 and 1 microg/ml, respectively. Meropenem was the most active compound against members of the family Enterobacteriaceae with MIC90s of < or =0.5 microg/ml. Against these species, GV129606 possessed activity superior to those of piperacillin, ceftazidime, cefpirome, and gentamicin, with MIC90s of < or =8 microg/ml, but its activity was two- to sixfold less than that of ciprofloxacin (with the exception of Proteus rettgeri and Providencia stuartii). Haemophilus spp., Moraxella catarrhalis, Neisseria gonorrhoeae, and Pseudomonas aeruginosa were also included in the spectrum of GV129606. GV129606 showed good antianaerobe activity, similar to metronidazole. It was stable against all clinically relevant beta-lactamases (similar to meropenem). The in vitro activity was confirmed in vivo against septicemia infections induced in mice by selected gram-positive and -negative bacteria with 50% effective doses (ED50s) of < or =0.05 and < or =0.5 mg/kg of body weight/dose, respectively. GV129606 was as effective as meropenem against septicemia in mice caused by ceftazidime-resistant Pseudomonas aeruginosa, exhibiting an ED50 of 0.33 mg/kg/dose. PMID- 9420051 TI - Influence of erythromycin resistance, inoculum growth phase, and incubation time on assessment of the bactericidal activity of RP 59500 (quinupristin dalfopristin) against vancomycin-resistant Enterococcus faecium. AB - RP 59500, a mixture of two semisynthetic streptogramin antibiotics (quinupristin and dalfopristin), is one of a few investigational agents currently in clinical trials with inhibitory activity against multiple-drug-resistant strains of Enterococcus faecium. We evaluated the bactericidal activity of this antimicrobial against 30 recent clinical isolates of vancomycin-resistant E. faecium, including 23 erythromycin-resistant (MIC, >256 microg/ml) and 7 erythromycin-intermediate (MIC, 2 to 4 microg/ml) strains. All isolates were inhibited by RP 59500 at 0.25 to 1.0 microg/ml. The bactericidal activity of RP 59500 was markedly influenced by the erythromycin susceptibility of the strains and by several technical factors, such as inoculum growth phase and time of incubation of counting plates. As determined by time-kill methods, RP 59500 at a concentration of 2 or 8 microg/ml failed to kill erythromycin-resistant organisms under any conditions. Bactericidal activity was observed against all seven erythromycin-intermediate isolates when log-phase inocula were used and the cells were counted after 48 h of incubation (mean reductions in viable bacteria for RP 59500 at concentrations of 2 and 8 microg/ml, 3.45 and 3.50 log10 CFU/ml, respectively), but killing was diminished when the plates were examined at 72 h (mean killing, 3.06 and 2.95 log10, CFU/ml, respectively). No bactericidal activity was observed when stationary-phase cultures were used. On the basis of these data, we expect that bactericidal activity of RP 59500 against the multiple drug-resistant E. faecium strains currently encountered would be distinctly uncommon. PMID- 9420052 TI - Antiviral drug susceptibility of human herpesvirus 8. AB - We studied the susceptibility of human herpesvirus 8 (HHV-8) to a number of antiherpesvirus agents. The acyclic nucleoside phosphonate (ANP) analogs cidofovir and HPMPA [(S)-1-(3-hydroxy-2-phosphonylmethoxypropyl)adenine] effected potent inhibition of HHV-8 DNA synthesis, with 50% effective concentrations (EC50) of 6.3 and 0.6 microM, respectively. Adefovir, an ANP with both antiretrovirus and antiherpesvirus activity, blocked HHV-8 DNA replication at a fourfold-lower concentration than did foscarnet (EC50 of 39 and 177 microM, respectively). The most potent inhibitory effect was obtained with the N-7 substituted nucleoside analog S2242 (EC50, 0.11 microM). The nucleoside analogs acyclovir, penciclovir, H2G ((R)-9-[4-hydroxy-2-(hydroxymethyl) butyl]guanine), and brivudine had weak to moderate effects (EC50 of > or =75, 43, 42, and 24 microM, respectively, and EC90 of > or =75 microM), whereas ganciclovir elicited pronounced anti-HHV-8 activity (EC50, 8.9 microM). PMID- 9420053 TI - Cloning and nucleotide sequence analysis of a gene encoding an OXA-derived beta lactamase in Acinetobacter baumannii. AB - A clinical strain of Acinetobacter baumannii (strain Ab41) that was resistant to all beta-lactam antibiotics tested except ceftazidime, ceftriaxone, ceftizoxime, and imipenem produced three beta-lactamases: a presumptive chromosomal cephalosporinase, a TEM-1-like beta-lactamase (pI 5.4), and a novel OXA-derived beta-lactamase named OXA-21 (pI 7.0). The gene encoding OXA-21 was located in an integron. The nucleotide sequence showed three mutations compared with the sequence of OXA-3, with two being silent; the nonsilent mutation generated a substitution of Ile-217 to Met. PMID- 9420054 TI - Comparison of in vitro antifungal susceptibilities of conidia and hyphae of filamentous fungi. AB - The MICs and minimum fungicidal concentrations (MFCs) of amphotericin B, fluconazole, ketoconazole, flucytosine, miconazole, and itraconazole for 12 isolates of filamentous opportunistic fungi (Scopulariopsis sp., Paecilomyces sp., Cladosporium spp., and Cladophialophora sp.) were determined by a broth microdilution method with hyphal and conidial inocula. With hyphal inocula MICs and MFCs were practically always substantially higher. Only 25% of the 60 MIC comparisons showed discrepancies of twofold or less, while the remaining comparisons showed much larger differences. PMID- 9420055 TI - Mutations at codon 184 in simian immunodeficiency virus reverse transcriptase confer resistance to the (-) enantiomer of 2',3'-dideoxy-3'-thiacytidine. AB - Variants of simian immunodeficiency virus (SIV) that display greater than 2,000 fold resistance to the (-) enantiomer of 2',3'-dideoxy-3'-thiacytidine (3TC) were generated through in vitro passage and drug selection. The polymerase regions of several of these resistant viruses were sequenced and were found to share either of two codon alterations at site 184 in reverse transcriptase (ATG to ATA [methionine to isoleucine] and ATG to GTA [methionine to valine]). The biological relevance of these substitutions for 3TC was confirmed by site-directed mutagenesis with the SIVmac239 infectious recombinant clone of SIV. PMID- 9420056 TI - Amino acid variation in the GyrA subunit of bacteria potentially associated with natural resistance to fluoroquinolone antibiotics. AB - In studies of genetic diversity in natural microbial populations, we have analyzed nucleotide sequences in the quinolone resistance-determining region of the bacterial gyrA gene in ciprofloxacin-resistant and nonselected soil bacteria obtained from the environment. It is apparent that this sequence is highly variable, and resistance to fluoroquinolone antibiotics occurring in environmental populations of bacteria is due at least in part to natural sequence variation in this domain. We suggest that the development of new antimicrobial agents, including completely synthetic antimicrobials such as the fluoroquinolones, should incorporate the analysis of resistance mechanisms among microbes in natural environments; these studies could predict potential mechanisms of resistance to be encountered in subsequent clinical use of the agents and would guide chemical modification designed to evade resistance development. PMID- 9420057 TI - Selection of multiple-antibiotic-resistant (mar) mutants of Escherichia coli by using the disinfectant pine oil: roles of the mar and acrAB loci. AB - Mutants of Escherichia coli selected for resistance to the disinfectant pine oil or to a household product containing pine oil also showed resistance to multiple antibiotics (tetracycline, ampicillin, chloramphenicol, and nalidixic acid) and overexpressed the marA gene. Likewise, antibiotic-selected Mar mutants, which also overexpress marA, were resistant to pine oil. Deletion of the mar or acrAB locus, the latter encoding a multidrug efflux pump positively regulated in part by MarA, increased the susceptibility of wild-type and mutant strains to pine oil. PMID- 9420058 TI - Canadian Multicenter Susceptibility Study, with a focus on cephalosporins, from 15 Canadian medical centers. The Canadian Multicenter Study Group. AB - We have previously reported on the in vitro susceptibilities of 4,482 microorganisms to 10 antimicrobial agents tested as part of a Canadian multicenter study. We now report on the remaining 10 agents tested in that study. Of the cephalosporins reported here, ceftriaxone had the greatest activity (82 to 100% susceptible isolates) against Enterobacteriaceae, compared to ceftizoxime (78 to 100%) and cefoperazone (78 to 100%). Cefoperazone activity against Pseudomonas aeruginosa was 87%, compared to 92% for ticarcillin-clavulanate. All agents had 97% or greater activity against Staphylococcus aureus. PMID- 9420059 TI - Sulfated carbohydrate compounds prevent microbial adherence by sexually transmitted disease pathogens. AB - Heparan sulfate (HS) serves as a receptor for adherence of herpes simplex viruses, Chlamydia trachomatis, Neisseria gonorrhoeae, and, indirectly, human immunodeficiency virus. Using primary human culture systems, we identified sulfated carbohydrate compounds that resemble HS and competitively inhibit infection by these pathogens. These compounds are candidates for intravaginal formulations for the prevention of sexually transmitted diseases. PMID- 9420060 TI - A recombinant retroviral system for rapid in vivo analysis of human immunodeficiency virus type 1 susceptibility to reverse transcriptase inhibitors. AB - We have developed a new recombinant retroviral system in which a library of infectious molecular clones of human immunodeficiency virus type 1 (HIV-1) is constructed with reverse transcriptase (RT) genes derived from viral RNA sequences in plasma. HIV-1 RT is amplified from plasma HIV-1 RNA by nested RT-PCR and cloned into a RT-defective HIV-1 proviral vector (xxLAI-np), generating 10(3) to 10(4) recombinant proviral clones from each reaction. The bulk cloning products or individual molecular clones are transfected into MT-2 cells to generate infectious virus. The resultant viruses are assayed for drug susceptibility in CD4+ cell lines to determine either the dominant phenotype of the recombinant virus mixture or the phenotypes of the individual viral clones. DNA sequencing of the cloned RT genes can identify mutations associated with phenotypic resistance of clonal mixtures or individual clones. This method can be used to rapidly detect the in vivo emergence of HIV-1 quasispecies resistant to RT inhibitors. PMID- 9420061 TI - Antipneumococcal activity of BAY 12-8039, a new quinolone, compared with activities of three other quinolones and four oral beta-lactams. AB - Activities of BAY 12-8039 against 205 pneumococci were tested by agar dilution. MICs (in micrograms per milliliter) at which 50 and 90% of the isolates are inhibited (MIC50s and MIC90s, respectively) were 0.125 and 0.25 (BAY 12-8039), 2.0 and 4.0 (ciprofloxacin and ofloxacin), and 0.25 and 0.5 (sparfloxacin). Beta lactam MIC50s and MIC90s for penicillin-susceptible, -intermediate, and resistant strains, in that order, were 0.016 and 0.03, 0.25 and 2.0, and 2.0 and 4.0 (amoxicillin); 0.03 and 0.06, 0.25 and 4.0, and 4.0 and 8.0 (ampicillin); 0.03 and 0.06, 0.5 and 4.0, and 4.0 and 8.0 (cefuroxime); and 0.03 and 0.125, 0.25 and 2.0, and 4.0 and 8.0 (cefpodoxime). At two times their MICs after 24 h, BAY 12-8039, ciprofloxacin, ampicillin, and cefuroxime were uniformly bactericidal (99.9% killing) against 12 strains; other compounds were bactericidal at four times their MICs. PMID- 9420062 TI - In vitro activities of new quinolones against Helicobacter pylori. AB - Compounds belonging to a new class of quinolones in which the fundamental C-6 fluorine atom was replaced were evaluated for in vitro antibacterial activity against 32 Helicobacter pylori strains. Since these substitutions resulted in higher inhibitory activities, these new desfluoroquinolones may be useful in eradicating H. pylori infections. PMID- 9420063 TI - Pharmacokinetics and safety of a single dose of stavudine (d4T) in patients with severe hepatic impairment. AB - This open-label study enrolled five subjects with biopsy-proven cirrhosis and moderate to severe hepatic impairment (Child-Pugh classification grade B or C) and five age- and gender-matched controls. All subjects received a single 40-mg oral dose of stavudine (d4T). Stavudine pharmacokinetics in subjects with hepatic impairment were similar to those in age- and gender-matched control subjects and were not substantially different from those previously observed in human immunodeficiency virus-infected patients. Based on these findings, stavudine use does not require modification of the dose or dosing interval for patients with liver disease. PMID- 9420064 TI - Single-dose pharmacokinetics of thalidomide in human immunodeficiency virus infected patients. AB - The pharmacokinetics of thalidomide in nine human immunodeficiency virus-infected patients were studied. Single doses of thalidomide were well absorbed, with mean peak concentrations (+/- standard deviations) of 1.17 +/- 0.21 and 3.47 +/- 1.14 microg/ml in the 100- and 300-mg dosing groups, respectively, and the mean elimination half-life was approximately 6 h. Adverse effects were mild, with drowsiness being reported for seven of nine patients. PMID- 9420065 TI - In vitro activities of doxycycline and enrofloxacin against European Chlamydia psittaci strains from turkeys. AB - The in vitro susceptibility of 14 European Chlamydia psittaci strains from turkeys to the antibiotics doxycycline and enrofloxacin was tested. For doxycycline the MIC ranged from 0.05 to 0.2 microg/ml, with an average of 0.1 microg/ml. For enrofloxacin the MIC was 0.25 microg/ml. Acquired resistance was not detected against doxycycline and enrofloxacin. PMID- 9420066 TI - Survival rates in patients with primary malignant brain tumors stratified by patient age and tumor histological type: an analysis based on Surveillance, Epidemiology, and End Results (SEER) data, 1973-1991. AB - OBJECT: The authors present population-based survival rate estimates for patients with malignant primary brain tumors based on an analysis of 18 years of data obtained from the Surveillance, Epidemiology, and End Results (SEER) program of the National Cancer Institute. METHODS: Estimates of survival rates at 2 and 5 years after diagnosis for patients with specific histological tumor types were categorized by patient's age at diagnosis (< or = 20 years, 21-64 years, and 65 years or older) and by the time period in which the patients were diagnosed (1973 1980, 1981-1985, 1986-1991). Where appropriate, survival estimates were adjusted for changing patterns in the mean age at diagnosis. CONCLUSIONS: The authors observed a pattern of declining survival rates in patients with increasing age of the patient at diagnosis for most histological groups and overall improvements in survival rates of patients across these time periods adjusting for age at diagnosis. There were improvements in 2- and 5-year survival rates over the three time periods for children and adults with medulloblastoma and for adults with astrocytoma and oligodendroglioma. Improvements in survival rates for pediatric patients with medulloblastoma have leveled off in the most recent time period, and gender differences in survival rates for patients with this tumor, which were present in the 1970s, have disappeared. Clinically significant improvements in survival rates were not apparent in patients aged 65 years and older. Changes in diagnostic and treatment procedures since the mid-1970s have resulted in improved survival rates for patients diagnosed as having medulloblastoma, oligodendroglioma, and astrocytoma, controlling for age at diagnosis. Glioblastoma multiforme continues to be the most intractable brain tumor. PMID- 9420067 TI - Demographics, prognosis, and therapy in 702 patients with brain metastases from malignant melanoma. AB - Brain metastases are a common and devastating complication in patients with malignant melanoma. Therapeutic options for these patients are limited, and the prognosis is usually poor. OBJECT: A retrospective review of 6953 patients with melanoma treated at a single institution was undertaken to identify demographic factors associated with the development of clinically significant brain metastases in 702 of these patients and to determine the factors influencing the prognosis of this population to permit more informed recommendations regarding surgical therapy. METHODS: Factors found to be associated with the development of brain metastases included male gender, primary lesions located on mucosal surfaces or on the skin of the trunk or head and neck, thick or ulcerated primary lesions, and histological findings of acral lentiginous or nodular lesions. The overall median survival time of all patients with brain metastases was 113.2 days, and these metastases contributed to the death of 94.5% of the patients in this group. Patients with primary lesions located in the head or neck region had a significantly shorter survival time relative to other patients with brain metastases, whereas patients with a single brain metastasis, patients without lung or multiple other visceral metastases, and patients whose initial presentation with melanoma included a brain metastasis had a significantly better prognosis. The small group of patients who survived for more than 3 years was characterized by the presence of a surgically treated, single brain metastasis in the absence of other visceral metastatic disease. CONCLUSIONS: Although most patients with brain metastases resulting from melanoma have a dismal prognosis, some who are likely to survive for longer periods can be identified. In these patients surgical resection can significantly prolong meaningful survival. The decision to recommend surgery should be based primarily on the resectability of the brain metastases and on the status and number of other organs with metastatic lesions. PMID- 9420068 TI - Bisegmental cervical interbody fusion using hydroxyapatite implants: surgical results and long-term observation in 70 cases. AB - Hydroxyapatite (HA) is the main constituent of bone mineral, and synthetic HA serves as a biocompatible and bioactive material. It permits bone growth on its surface and forms a union with the adjacent bone. OBJECT: The authors have developed implants made of porous HA, which they have used in more than 90 cases in the past 6 years to achieve cervical interbody fusion. The implants were designed to provide maximum durability, biomechanical stability, and alignment preservation and to be technically easy to use. The authors summarize their experience and results with the use of these implants. METHODS: The results of postoperative follow-up observation of 12 months or longer (mean 37.1 +/- 2.4 months) are available in 70 patients with underlying disease including: spondylosis, disc extrusion, ossification of the posterior longitudinal ligament (PLL), hypertrophy of the PLL, and trauma. The patients' ages at the time of surgery ranged from 22 to 83 years (mean 50.6 +/- 1.3 years). Flexion-extension radiographs and tomograms, obtained 6 and 12 months after surgery and every year thereafter, were used to demonstrate solid fusion in all cases. Dislocation of the implant occurred in three patients who were treated during the early portion of the series. At 6 to 12 months after surgery, encasement of the implant and formation of union were observed. Normal lordosis, if present prior to surgery, was maintained postsurgery. No neurological deterioration related to the site of fusion occurred during the period of observation. CONCLUSIONS: The authors conclude that satisfactory interbody fusion can be achieved by using HA implants, provided their design is appropriate and adequate surgical techniques are used. PMID- 9420069 TI - Pulmonary function and radiographic abnormalities related to neurological outcome after aneurysmal subarachnoid hemorrhage. AB - OBJECT: This observational study is based on a consecutive series of 207 patients with aneurysmal subarachnoid hemorrhage who were treated within 7 days of their most recent bleed. The purpose of the study was to evaluate the effect of respiratory failure on neurological outcome. METHODS: Pulmonary function was assessed by determination of parameters describing pulmonary oxygen transport and exchange, by using composite scores for quantification of lung injury (lung injury score [LIS]) and mechanical ventilator settings (PIF score). Pulmonary function was related to the Hunt and Hess (H & H) grade assigned to the patient at hospital admission (p < 0.001). The pattern and time course of lung injury differed significantly between patients with H & H Grade I or II, Grade III, and Grade IV or V. Hunt and Hess grade, Fisher computerized tomography grade, intracranial pressure, cerebral perfusion pressure, LIS, ratio of PaO2 to the fraction of inspired oxygen (FiO2), and the ratio of the alveolar-minus-arterial oxygen tension difference (AaDO2) to FiO2 were related to neurological outcome (p < 0.001). The LIS on the day of maximum lung injury remained an independent predictor of outcome (p = 0.01) in a stepwise logistic regression analysis. The probability of poor neurological outcome significantly increased with both decreasing cerebral perfusion pressure and increasing severity of lung injury. CONCLUSIONS: The overall mortality rate was 22.2% (46 of 207 patients). Subarachnoid hemorrhage and its neurological sequelae accounted for the principal mortality in this series. Medical (nonneurological and nontreatment-related) complications accounted for 37% of all deaths. Systemic inflammatory response syndrome with associated multiple organ dysfunction syndrome was the leading cause of death from medical complications. The authors conclude that respiratory failure is related to neurological outcome, although it is not commonly the primary cause of death from medical complications. PMID- 9420070 TI - Increase in diameters of vasospastic intracranial arteries by intraarterial papaverine administration. AB - OBJECT: This study was conducted to determine if there is a change in intracranial arterial diameters after papaverine infusion for vasospasm and to determine whether the change occurs in proximal, intermediate, and distal arteries. METHODS: The authors measured arterial diameters retrospectively in all patients who received intraarterial papaverine for treatment of vasospasm between November 1992 and August 1995. Patients who received papaverine in the same session with or following angioplasty were excluded. Measurements were made in a blinded manner with the aid of a magnification loupe at 12 predetermined sites on each angiogram before and after papaverine infusion. Eighty-one treatments in 34 patients were included. Angiograms obtained at the time of presentation with subarachnoid hemorrhage (SAH) were examined in 26 of the 34 patients. Nine carotid territories visualized by repeated angiography on the day after infusion were examined to determine the duration of the papaverine effect. CONCLUSIONS: In all treatment groups an increase was found in the average arterial diameters ranging from 2.8 to 73.9%, with a mean increase of 26.5%. Increases in diameter were observed in proximal, intermediate, and distal arteries. The timing of treatments ranged from Day 3 to Day 19 post-SAH, and there was no relationship between timing and arterial responsiveness (r = -0.06). There was a moderately good correlation between the degree of vasospasm in an artery and its responsiveness to papaverine (r = -0.54, -0.66, and -0.66, for proximal, intermediate, and distal arteries, respectively). The effect of papaverine did not persist until the following day in patients in whom repeated angiography was performed. PMID- 9420071 TI - Dose-response tolerance of the visual pathways and cranial nerves of the cavernous sinus to stereotactic radiosurgery. AB - As the number of patients treated with stereotactic radiosurgery increases, it becomes particularly important to define with precision adverse effects on distinct structures of the nervous system. OBJECT: This study was designed to assess the dose-response tolerance of the visual pathways and cranial nerves after exposure of the cavernous sinus to radiation. METHODS: A total of 66 sites in the visual system and 210 cranial nerves of the middle cranial fossa were investigated in 50 patients who had undergone gamma knife treatment for benign skull base tumors. The mean follow-up period was 40 months (range 24-60 months). Follow-up examinations consisted of neurological, neuroradiological, and neuroophthalmological evaluations. The actuarial incidence of optic neuropathy was zero for patients who received a radiation dose of less than 10 Gy, 26.7% for patients receiving a dose in the range of 10 to less than 15 Gy, and 77.8% for those who received doses of 15 Gy or more (p < 0.0001). Previously impaired vision improved in 25.8% and was unchanged in 51.5% of patients. No sign of neuropathy was seen in patients whose cranial nerves of the cavernous sinus received radiation doses of between 5 and 30 Gy. Because tumor control appeared to have been achieved in 98% of the patients, the deterioration in visual function cannot be attributed to tumor progression. CONCLUSIONS: The structures of the visual pathways (the optic nerve, chiasm, and tract) exhibit a much higher sensitivity to single-fraction radiation than other cranial nerves, and their particular dose-response characteristics can be defined. In contrast, the oculomotor and trigeminal nerves have a much higher dose tolerance. PMID- 9420072 TI - Formation of intracerebral cavernous malformations after radiation treatment for central nervous system neoplasia in children. AB - OBJECT: Radiation is a common treatment modality for pediatric brain tumors. The authors present a retrospective review of six children who developed cerebral cavernous malformations after they underwent radiation treatment for central nervous system (CNS) neoplasia and propose two possible models to explain the formation of cavernous malformations. METHODS: Three boys, aged 13, 9, and 17 years, suffered intracerebral hemorrhages from cerebral cavernous malformations 87, 94, and 120 months, respectively, after they received whole-brain radiation therapy (WBRT) for acute lymphocytic leukemia. A 10-year-old girl and a 19-year old man developed temporal lobe cavernous malformations 46 and 48 months, respectively, after they received radiation therapy for posterior fossa astrocytomas. A 12-year-old girl developed a temporal lobe cavernous malformation 45 months after WBRT was administered for a medulloblastoma. In all of these cases the cavernous malformation appeared in the irradiated field, was not known to be present prior to radiation therapy, and developed after a latency period following treatment. The incidence of cavernous malformations in these patients suggests that children who undergo radiation therapy of the brain may have an increased risk of hemorrhage. CONCLUSIONS: Two possible models may explain the formation of cavernous malformations following brain radiation in these patients. First, the cavernous malformations may form de novo in response to the radiation. Second, the cavernous malformations may have been present, but radiographically occult, at the time of radiation therapy and may have hemorrhaged in response to the radiation. The authors conclude that cavernous malformations may develop after brain radiation and propose a possible mechanism for this formation. PMID- 9420073 TI - Split spinal cord malformations in children. AB - OBJECT: The authors reviewed and analyzed information on 74 patients with split spinal cord malformations (SSCMs) treated between January 1, 1980 and December 31, 1996 at their institution with the aim of defining and classifying the malformations according to the method of Pang, et al. METHODS: Computerized tomography myelography was superior to other radiological tools in defining the type of SSCM. There were 46 girls (62%) and 28 boys (38%) ranging in age from less than 1 day to 12 years (mean 33.08 months). The mean age (43.2 months) of the patients who exhibited neurological deficits and orthopedic deformities was significantly older than those (8.2 months) without deficits (p = 0.003). Fifty two patients had a single Type I and 18 patients a single Type II SSCM; four patients had composite SSCMs. Sixty-two patients had at least one associated spinal lesion that could lead to spinal cord tethering. After surgery, the majority of the patients remained stable and clinical improvement was observed in 18 patients. CONCLUSIONS: The classification of SSCMs proposed by Pang, et al., will eliminate the current chaos in terminology. In all SSCMs, either a rigid or a fibrous septum was found to transfix the spinal cord. There was at least one unrelated lesion that caused tethering of the spinal cord in 85% of the patients. The risk of neurological deficits resulting from SSCMs increases with the age of the patient; therefore, all patients should be surgically treated when diagnosed, especially before the development of orthopedic and neurological manifestations. PMID- 9420074 TI - Induction chemotherapy followed by reduced-volume radiation therapy for newly diagnosed central nervous system germinoma. AB - OBJECT: Although curative, radiation, which is conventionally administered for germinomas, causes significant neurological sequelae. This study aimed at reducing the volume and dose of radiation to a localized level of 24 Gy by pretreating the patient with chemotherapy. METHODS: Seventeen patients were divided into two risk groups based on serological findings and the extent of tumor. They were treated with chemotherapy prior to receiving localized radiation therapy. Six patients with solitary pure germinomas were treated with three or four cycles of cisplatin and etoposide (EP regimen) followed by 24-Gy local radiation therapy. Eleven patients with human chorionic gonadotropin (HCG) secreting, multifocal, or disseminated germinomas received four to five cycles of ifosfamide, cisplatin, and etoposide (ICE regimen) followed by 24-Gy local radiation therapy. Craniospinal ports were used only in three cases of germinomas with dissemination. Gross-total resection was performed in three patients. Fourteen patients were able to be evaluated for their responses to chemotherapy. All patients achieved a complete response within three cycles. At a median follow up duration of 24 months, 16 patients (94%) were alive without recurrence. One patient with an HCG-secreting germinoma experienced recurrence 38 months after surgery. That patient underwent successful salvage therapy using the same protocol. Thus, all 17 patients became free of disease with a 70 to 100% Karnofsky Performance Scale status. Toxicities associated with this study's chemotherapy regimen were mostly transient. No patient showed neurological or endocrinological deterioration during the follow-up period. CONCLUSIONS: The EP and ICE regimens were highly effective in treating the central nervous system germinomas and permitted dose and volume reduction of the radiotherapy. Localized 24-Gy irradiation was sufficient for disease control. PMID- 9420075 TI - Infusion of intrathecal baclofen for generalized dystonia in cerebral palsy. AB - Generalized dystonia occurs in 15 to 25% of persons with cerebral palsy (CP) and responds poorly to medical and surgical treatments. OBJECT: After the authors observed a woman whose dystonic CP was dramatically improved by continuous infusion of intrathecal baclofen, they designed this pilot study to evaluate the effect of this treatment on a group of patients with dystonic CP. METHODS: The authors assessed the short-term response to intrathecal baclofen infusion in 12 patients with dystonic CP. An intrathecal catheter was inserted percutaneously and connected to an external microinfusion pump. The infusion began at a rate of 100 microg/day and was increased by 50 microg every 12 hours until the dystonia abated, adverse effects occurred, or the dose reached 900 microg/day with no improvement. Two observers, one blinded and one not blinded to the patient's treatment status, viewed videotapes made before and after the infusions and graded the dystonia in eight body regions, using a 5-point scale. Overall and regional scores were compared by using Wilcoxon signed-rank tests. CONCLUSIONS: Dystonia diminished in 10 of 12 patients whose average daily dose of intrathecal baclofen was 575 microg. Overall dystonia scores and scores for the extremities, trunk, and cervical regions were significantly better after infusion (p = 0.003). The two observers' scores were not significantly different. Programmable infusion pumps were subsequently implanted in eight patients for long-term therapy and improvement was sustained in six (p < 0.05). Intrathecal baclofen infusion is a promising treatment option for generalized dystonia associated with CP. The effects of intrathecal baclofen infusion on dystonia can be evaluated by using short-term continuous infusions. PMID- 9420076 TI - Endoscopic craniectomy for early surgical correction of sagittal craniosynostosis. AB - OBJECT: The authors sought to minimize scalp incisions, blood loss, and operative time by using endoscopically assisted strip craniectomies and barrel-stave osteotomies to treat infants with sagittal suture synostosis. METHODS: Four patients, aged 2, 4, 9, and 12 weeks, who presented with scaphocephaly underwent endoscopic midline craniectomies through small midline scalp incisions. The mean operative time for the procedure was 1.68 hours (range 1.15-2.8 hours); the mean blood loss was 54.2 ml (range 12-150 ml). Three patients did not require blood transfusions and were discharged within 24 hours. Postoperatively, all patients were fitted with custom cranial molding helmets. Follow-up evaluation ranged between 8 and 15 months. All patients had successful correction of their scaphocephaly with no mortalities, morbidities, or complications. CONCLUSIONS: The use of endoscopic techniques for early correction of sagittal synostosis is safe; decreases blood loss, operative time, and hospitalization costs; and provides excellent early surgical results. PMID- 9420077 TI - Laser-assisted neuroendoscopy using a neodymium-yttrium aluminum garnet or diode contact laser with pretreated fiber tips. AB - OBJECT: Although lasers have proved to be valuable in neuroendoscopy, surgeons are still not comfortable using high-energy laser endoscopic probes in proximity to vital structures such as the basilar artery in third ventriculostomy. The authors have developed a special laser catheter for use in neuroendoscopy; the object of this paper is to present their experimental and clinical experiences using the catheter. METHODS: This laser catheter is fitted with an atraumatic ball-shaped fiber tip that is pretreated with a layer of carbon particles. These carbon particles absorb approximately 90% of the energy emitted, which is very effectively converted into heat. As the heat is generated in this very thin layer of carbon coating, the temperature at the surface of the ball-shaped tip reaches ablative temperatures instantly at powers of only a few watts per second, which has enabled the authors to limit drastically the amount of laser light used and the length of exposure needed, thereby increasing safety even around critical structures. CONCLUSIONS: The authors present experimental data and their clinical experience using these pretreated fiber tips with a neodymium-yttrium aluminum garnet contact laser or a diode contact laser in 49 patients (22 males and 27 females) and a variety of procedures: third ventriculocistemostomy (33 patients), cyst fenestration (nine patients), colloid cyst resection (six patients), and fenestration of the septum pellucidum (one patient). There was no instance of mortality or increased morbidity. To date, the procedure success rate is 100% and the overall outcome success rate is 86%. The authors conclude that pretreated atraumatic ball-shaped fiber tips now make laser application safe and effective in a variety of neuroendoscopic procedures. Because of their low power range (only several watts), compact diode lasers will be the energy source of first choice. PMID- 9420078 TI - Cell type-specific development of rodent central nervous system progenitor cells in culture. AB - OBJECT: The aim of the study was to assess the pluripotential central nervous system (CNS) progenitor cells that give rise to the many differentiated neuronal and glial cell types of the adult mammalian brain and the role of peptide growth factors such as the epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF). The action of these factors is crucial to the survival and ultimate differentiation of these CNS progenitor cells. However, the precise role of EGF and bFGF in the time course of cellular development, the acquisition of cell type specificity, and possible differential mitogenic behavior has not been clearly defined. METHODS: The authors defined the time course of CNS progenitor cell development in cultured embryonic rodent cells by using immunocytochemical analysis to identify the expression of pluripotential (nestin)-, neuron (microtubule-associated protein-2 [MAP-2])-, and glia (glial fibrillary acidic protein [GFAP])-specific proteins in response to treatment with EGF and bFGF alone or in combination. The bromodeoxyuridine (BUdR) labeling index for each treatment group was used to define the mitogenic effects of each growth factor. In this investigation, the authors observed that progenitor cells develop in a stereotypical fashion when exposed to bFGF or EGF. Marked staining for nestin was evident soon after plating. This declined over time as staining for MAP-2 and GFAP increased. When treated with EGF alone, cells maintained their nestin immunoreactivity longer than those treated with bFGF alone or in combination with EGF. Treatment with bFGF alone promoted a significant increase in MAP-2 and, to a much lesser extent, GFAP reactivity. This was observed concomitant with the decline in nestin staining. The BUdR labeling index was similar among the different treatment groups and declined similarly over time in all treatment groups. CONCLUSIONS: The effects of EGF and/or bFGF on the expression of development- and lineage-specific markers likely reflect the specific effects of these factors on developmental processes. These data indicate that bFGF exerts a preferential effect on neuronal development and, to a lesser extent, glial development, which is not explained by selective mitogenicity. The persistence of nestin staining seen in the cells treated with EGF alone indicates that EGF may function as a stem cell survival factor. This study provides evidence that CNS cell type-specific development can be altered by the manipulation of peptide growth factors that act as differentiation agents. PMID- 9420079 TI - Transfection of C6 glioma cells with the bax gene and increased sensitivity to treatment with cytosine arabinoside. AB - OBJECT: Genes known to be involved in the regulation of apoptosis include members of the bcl-2 gene family, such as inhibitors of apoptosis (bcl-2 and bcl-xl) and promoters of apoptosis (bax). The authors investigated a potential approach for the treatment of malignant gliomas by using a gene transfection technique to manipulate the level of an intracellular protein involved in the control of apoptosis. METHODS: The authors transfected the murine bax gene, which had been cloned into a mammalian expression vector, into the C6 rat glioma cell line. Overexpression of the bax gene resulted in a decreased growth rate (average doubling time of 32.96 hours compared with 22.49 hours for untransfected C6, and 23.11 hours for clones transfected with pcDNA3 only), which may be caused, in part, by an increased rate of spontaneous apoptosis (0.77 +/- 0.15% compared with 0.42 +/- 0.08% for the vector-only transfected C6 cell line; p = 0.038, two tailed Student's t-test). Treatment with 1 microM cytosine arabinoside (ara-C) resulted in significantly more cells undergoing apoptosis in the cell line overexpressing bax than in the vector-only control cell line (23.57 +/- 2.6% compared with 5.3 +/- 0.7% terminal deoxynucleotidyl transferase-mediated biotinylated-deoxyuridine triphosphate nick-end labeling technique-positive cells; p = 0.007). Furthermore, measurements of growth curves obtained immediately after treatment with 0.5 microM ara-C demonstrated a prolonged growth arrest of at least 6 days in the cell line overexpressing bax. CONCLUSIONS: These results can be used collectively to argue that overexpression of bax results in increased sensitivity of C6 cells to ara-C and that increasing bax expression may be a useful strategy, in general, for increasing the sensitivity of gliomas to antineoplastic treatments. PMID- 9420080 TI - Transparaspinal exposure of dumbbell tumors of the spine. Report of two cases. AB - The authors present a surgical technique for resection of dumbbell tumors of the spine. The transparaspinal exposure combines laminectomy and sectioning of the paraspinal muscles through a transverse incision. The procedure allows total tumor resection by means of a single posterior approach in selected patients, thus obviating the need for a combined anteroposterior operation. The advantages and disadvantages of the transparaspinal approach compared with the more extensive lateral extracavitary approach are discussed. PMID- 9420081 TI - Multiple radiation-induced intracranial lesions after treatment for pituitary adenoma. Case report. AB - This 53-year-old man presented with a syncopal episode 31 years after undergoing craniotomy and external-beam radiation for a pituitary macroadenoma. A gadolinium enhanced magnetic resonance (MR) image of the brain demonstrated a 2.5-cm enhancing mass in the right caudate region that had not been seen on previous studies. A stereotactically guided biopsy procedure was performed to obtain specimens from the mass, which were consistent with ependymoma. The MR image also revealed two additional lesions that appeared to be within the radiation fields: a right temporal meningioma and a left frontal cavernous malformation. A review of the literature found three previous reports in which ependymomas presented after radiation therapy. PMID- 9420082 TI - Meningeal melanocytoma. Report of a case and a historical comparison. AB - Meningeal melanocytomas are rare tumors of the central nervous system that are found almost exclusively in the posterior fossa and spinal cord and whose natural history is poorly defined. In this report, the authors review the clinical presentation, radiological appearance, operative findings, and histological features in two cases of meningeal melanocytoma: one cranial and one spinal. Two women, aged 21 and 30 years, were admitted to the hospital 60 years apart: the first because of progressive paraplegia and the second because of slowly progressive hearing loss. The first patient had an extradural tumor that was treated by laminectomy, subtotal resection, and postoperative radiotherapy in 1936. Her symptoms recurred 16 years later and she underwent reoperation of the residual tumor, which was found to have an intradural component. The authors' patient, who presented 60 years later, underwent plain and enhanced computerized tomography and magnetic resonance imaging that demonstrated a large posterior fossa lesion indicative of either an acoustic neuroma or a meningioma. She underwent posterior fossa decompression but only partial excision of the tumor could be accomplished because vigorous bleeding limited the extent of the resection. Surgery was followed by radiotherapy. The residual tumor enlarged despite these measures and required repeated resection 6 months later. At the second operation the tumor was much less vascular, perhaps reflecting the effects of radiotherapy, and was removed almost entirely. The patient died 6 months later from an anticoagulant-related cerebellar hemorrhage. In both cases the lesions were jet black, and histological examination revealed melanin-containing hypercellular tumors with rare mitotic figures. Meningeal melanocytomas are being diagnosed with increased frequency in parallel with improvements in neuroimaging and clarification of histological features. Clinical presentation of patients with these tumors typically occurs in their fifth decade and women are affected twice as often as men. The posterior fossa lesions can mimic acoustic neuromas and meningiomas in location and radiological appearance; however, the internal auditory canal is normal. In the spine, meningeal melanocytomas present with the clinical features of myeloradiculopathy. Diagnosis is made intraoperatively from the gross, jet-black appearance of the tumor and from histological examination. Vascularity, size, and location may render complete resection unfeasible. Because of the tumor's propensity to recur, radiotherapy has been recommended but its role remains to be elucidated. PMID- 9420083 TI - Intradural neurotropic spread of malignant mesothelioma. Case report and review of the literature. AB - This 54-year-old man with a history of right-sided malignant mesothelioma presented with signs of a partial spinal cord syndrome. The tumor had invaded the lower trunk of the brachial plexus and spread along the T-1 nerve root beneath the arachnoid onto the spinal cord itself. Mesothelioma, despite its known predilection for local spread, is rarely encountered within the spinal canal. Neurotropism is commonly encountered in facial malignancies; however, it has never been reported to affect the brachial plexus and spinal cord. PMID- 9420084 TI - Germinoma causing wallerian degeneration. Case report and review of the literature. AB - Germinomas occurring in the thalamus and basal ganglia sometimes cause atrophy of the cerebral hemisphere on the affected side. The authors present the case of a 12-year-old girl with a germinoma that developed in the basal frontal lobe and cerebral basal ganglia. Magnetic resonance imaging showed atrophy not only of the cerebrum but also of the brainstem. A T2-weighted image revealed an area of high intensity that proved to be wallerian degeneration extending from the corona radiata and internal capsule to the brainstem. The authors suggest that this pathological change may be involved in the development of the symptoms and hemiatrophy associated with germinomas in this region of the brain. PMID- 9420085 TI - Diagnostic and microsurgical presentation of intracranial angiolipomas. Case report and review of the literature. AB - Angiolipomas (ALs) are hamartomas composed of abnormally differentiated vessels and mature adipose tissue. Although they are most commonly found in peripheral tissues, ALs sometimes grow in the spinal epidural space. Intracranial ALs (ICALs) are rare: only seven cases have been reported in the literature. The authors describe the case of a 70-year-old woman who presented with ocular symptoms from a clinically and radiologically progressing parasellar ICAL. The radiological as well as the microsurgical findings are illustrated and compared with the seven previously published cases. The most frequent location of ALs is the skull base, especially the parasellar region. Other ICALs were diagnosed as components of cerebral arteriovenous malformations and were not symptomatic by themselves. Neuroradiological studies of ICALs usually demonstrate the characteristics of both adipose and vascular tissues. However, a review of the literature shows that the diagnosis had not been suspected preoperatively in any of the cases. Operative descriptions emphasize that most neurosurgeons were caught off guard by the profuse bleeding and the unusual relationship of this unexpected lesion to the cavernous sinus, so that removal was rarely complete. The authors conclude that preoperative diagnosis of ICALs is achievable based on magnetic resonance analysis, which should help optimize the microsurgical management of these lesions. PMID- 9420086 TI - Multiple appearing and vanishing aneurysms: primary angiitis of the central nervous system. Case report. AB - The authors present the case of a patient with ischemic episodes and recurrent intracerebral hemorrhages probably caused by primary angiitis of the central nervous system (CNS). An initial angiogram revealed multiple cerebral artery aneurysms as well as vascular wall irregularity; a second angiogram obtained 2 years later, however, did not demonstrate the previous aneurysms but instead showed new ones together with stenosis. Based on the histopathological findings and clinical course in this case, the multiple aneurysms appear to have been induced by vascular wall fragility and subsequent self-repair resulting from primary angiitis of the CNS. The authors present the histological and clinical characteristics of this unusual case of granulomatous, necrotizing CNS vasculitis. PMID- 9420087 TI - Large glial cyst of the pineal gland: a possible growth mechanism. Case report. AB - The authors report on a patient who presented with a large symptomatic glial cyst of the pineal gland communicating with the third ventricle. The hole between the ventricle and the cyst, suspected on magnetic resonance images, was found at surgery. The to-and-fro flow mechanism is considered to be involved in the pathogenesis of growth in this unusual large glial cyst of the pineal gland, although this mechanism cannot be applied universally. PMID- 9420088 TI - Stereotactic brachytherapy for a cystic metastatic brain tumor in the midbrain. Case report. AB - The authors report a rare case of a cystic metastasis in the midbrain that was successfully treated by brachytherapy following stereotactic biopsy and aspiration of the intratumoral cyst. Stereotactic aspiration of cystic lesions can lead to clinical improvement and brachytherapy prevents cyst recurrence. A 46 year-old man was referred to the authors' institution with a 2-month history of a left hemisensory disturbance and a 1-month history of progressive hemiparesis. Magnetic resonance (MR) imaging revealed a ring-enhancing cystic mass in the midbrain. On the basis of this imaging study, a differential diagnosis that included brainstem abscess, glioma, and metastatic tumor was made. Magnetic resonance imaging-guided stereotactic biopsy and aspiration of the intratumoral cyst were performed, yielding 5 ml of yellowish-white fluid. Histological examination provided a diagnosis of adenocarcinoma. During the surgery, a catheter through which brachytherapy would be delivered was inserted at a predetermined target. The patient's left hemiparesis and sensory disturbance were markedly improved and brachytherapy was begun 2 days postoperatively. Three radioactive isotopes composed of iridium-192 were implanted to irradiate the tumor tissue. The total dose at the tumor periphery was 30 Gy, which was administered over 100 hours. External-beam radiotherapy (20 Gy) was added after completion of the brachytherapy. At discharge from the hospital, the patient was alert and all his neurological symptoms had resolved. Follow-up MR imaging revealed stabilization of the cyst and no recurrence of the tumor. The patient is alive and well 18 months following the brachytherapy. This case suggests that brachytherapy can delay cyst recurrence, suppress tumor growth, and prolong survival in patients with cystic brainstem metastasis. PMID- 9420089 TI - Giant lateral sinus pericranii. Case report. AB - A case of giant lateral sinus pericranii, which presented in a patient during early childhood as a soft, collapsible mass and gradually grew until it reached 13 x 9 cm when the patient was 36 years of age, is reported. The patient underwent successful surgery and the lesion was totally excised. The results of diagnostic tests (computerized tomography scanning, magnetic resonance imaging, cerebral angiography, and sinusography) and surgery-related problems are presented and discussed. PMID- 9420090 TI - Disc herniation at T1-2. Report of four cases and literature review. AB - In preparing this paper, the authors reviewed their experiences with four cases of T1-2 disc herniation as well as the medical literature on the subject. Intervertebral thoracic disc herniations are uncommon and high thoracic disc herniations are rare. In the upper third of the thoracic spine, T1-2 is the most common level for disc ruptures. Four cases of disc herniation at T1-2 that caused T-1 radiculopathy are reported in this paper. In reviewing the literature on thoracic disc herniation, the authors found 27 cases at the T1-2 level, 23 of which were lateral disc herniations that produced radiculopathy and four of which were central disc herniations that caused myelopathy. The clinical signs and symptoms of T-1 radiculopathy are similar to those of C-8 radiculopathy; however, distinguishing features can frequently be found on neurological examination. The T-1 radiculopathy usually involves weakness of the intrinsic muscles of the hand. The motor deficit of C-8 radiculopathy involves the intrinsic muscles of the hand and most of the flexors and extensors of the fingers and wrist. The T-1 radiculopathy may produce Horner's syndrome (oculosympathetic paralysis) and diminished sensation in the axilla, which are not found with C-8 radiculopathy. In clinical presentation as well as in treatment, the lateral T1-2 disc herniation resembles a cervical disc herniation, whereas the central T1-2 disc herniation displays the usual appearance of a thoracic disc herniation. PMID- 9420091 TI - Thoracic duct injury during anterior cervical discectomy: a rare complication. Case report. AB - Chylous fistula resulting from intraoperative injury to the cervical thoracic duct is well described as a complication of neck dissection. However, injury to the thoracic duct during spinal surgery is rarely reported. The authors present the first case of thoracic duct injury occurring during cervical discectomy and fusion via an anterior approach. The anomalous location of the terminal arch of the thoracic duct in this patient contributed to the complication. The morbidity of chyle leakage is minimized by its early recognition, a thorough understanding of lymphatic system anatomy, and aggressive management of the thoracic duct injury. PMID- 9420092 TI - Postlaminectomy cervical spinal cord compression demonstrated by dynamic magnetic resonance imaging. Case report. AB - This 32-year-old man had undergone C3-7 laminectomy for posttraumatic cervical myelopathy associated with spinal canal stenosis. He developed recurrent myelopathy 5 years after the initial operation. Dynamic magnetic resonance (MR) imaging of the cervical spine demonstrated spinal cord compression with diffuse canal stenosis while the neck was in the extended position, whereas no significant stenosis was visualized in the neutral position. Sagittal and axial MR images of the affected levels demonstrated striking changes in the cervical spinal cord configuration. Because of an associated hard osteophyte formation and protruded disc, as well as a hypertrophied posterior longitudinal ligament, an anterior decompression and fusion with plate fixation were performed from C-4 to C-7. The postoperative course was uneventful, with subsequent neurological improvement. It is concluded that dynamic MR imaging aids the search for the cause of recurrent postlaminectomy cervical myelopathy after initial improvement following decompressive surgery. PMID- 9420093 TI - Failure of transodontoid screw fixation. Case report. AB - Anterior odontoid screw fixation is being performed with increasing frequency and may currently be the treatment of choice for Type II and selected Type III odontoid fractures, because it is the only surgical fusion that preserves C1-2 motion. Typically patients are immobilized postoperatively in a simple cervical collar. The authors present a case of postoperative fracture of the anterior body of the axis secondary to screw dislocation 5 weeks after single anterior odontoid screw osteosynthesis. Possible reasons for this rare complication and its implications for the technique are discussed. PMID- 9420094 TI - Akinetic mutism and magnetic resonance imaging in obstructive hydrocephalus. Case illustration. PMID- 9420095 TI - Arthur Roland Elvidge (1899-1985): contributions to the diagnosis of brain tumors and cerebrovascular disease. AB - The contributions of Arthur Elvidge (1899-1985), Wilder Penfield's first neurosurgical recruit, to the development of neurosurgery have been relatively neglected, although his work in brain tumors extended the previous work of Percival Bailey and Harvey Cushing. He published rigorous correlations of clinical and histological information and formulated a revised, modern nosology for neuroepithelial tumors, including a modern histological definition of glioblastoma multiforme. Well ahead of his time, he believed that glioblastoma was not strictly localized and was the first to comment that the tumor frequently showed "satellitosis." He was the first neurosurgeon in North America to use angiography as a radiographic aid in the diagnosis of cerebrovascular disease. Having studied with Egas Moniz, he was the first to detail the use of angiographic examinations specifically for demonstrating cerebrovascular disorders, believing that it would make possible routine surgery of the intracranial blood vessels. Seeking to visualize all phases of angiography, he was the impetus behind the design of one of the first semi-automatic film changers. Elvidge and Egas Moniz made the first observations on thrombosis of the carotid vessels independently of each other. Elvidge elucidated the significance of embolic stroke and commented on the ischemic sequelae of subarachnoid hemorrhage. Besides his contributions to neurosurgery, he codiscovered the mode of transmission of poliomyelitis. Elvidge's soft-spoken manner, his dry wit and candor, mastery of the understatement, love of exotic travel, and consummate dedication to neurosurgery made him a favorite of patients, neurosurgery residents, nurses, and other hospital staff. His accomplishments and example as teacher and physician have become part of neurosurgery's growing legacy. PMID- 9420096 TI - Academic chairmen. PMID- 9420097 TI - Dural inversion for moyamoya disease. PMID- 9420098 TI - Effects of exogenous cytokines on intravascular clearance of bacteria in normal and splenectomized mice. AB - BACKGROUND: Pretreatment with interleukin-1 (IL-1), granulocyte colony stimulating factor (G-CSF), and granulocyte macrophage colony-stimulating factor (GM-CSF) can improve alveolar macrophage bactericidal activity against pneumococci. These effects vary in eusplenic and asplenic mice. Likewise, these cytokines have been shown to improve survival after an aerosol pneumococcal challenge. Mice dying in these studies had positive blood cultures and disseminated infection. The purpose of this study was to determine the effect of cytokine pretreatment on intravascular clearance of bacteria from eusplenic and asplenic mice. METHODS: Two weeks after splenectomy or sham operation, mice were pretreated for various times with IL-1, G-CSF, or GM-CSF or their corresponding vehicles. Mice then received tail-vein injections of bacteria (0.1 mL), and quantitative blood cultures were performed 15 and 30 minutes thereafter. RESULTS: Splenectomized mice had impaired clearance of both pneumococci and Pseudomonas compared with sham-operated mice (p < 0.05). IL-1 enhanced clearance in splenectomized mice (p < 0.001) but not in sham-operated mice (p not significant). G-CSF enhanced bacterial clearance in sham-operated mice (p < 0.01) but not in splenectomized mice (p not significant). GM-CSF enhanced clearance in both groups (p < 0.001). CONCLUSION: The net effects of exogenous cytokine therapy for infections depends on the state of the host defenses at the time of therapy. These agents may be useful as adjuvants for the treatment of infections, but further study is warranted. PMID- 9420099 TI - Regulation of whole blood tumor necrosis factor production upon endotoxin stimulation after severe blunt trauma. AB - BACKGROUND: Trauma has been recognized to be accompanied by alterations of leukocyte functions such as cytokine release. The regulatory principles involved in these changes are still poorly defined. To further characterize leukocyte function after multiple trauma, endotoxin-stimulated tumor necrosis factor (TNF) production of trauma patients' whole blood and a possible regulatory mechanism were studied. METHODS: Endotoxin responsiveness in trauma patients (n = 18, Injury Severity Score = 24 +/- 7) was assayed ex vivo using a whole blood model. TNF release and TNFalpha mRNA levels were determined during a 14-day period. Furthermore, the influence of patients' sera on whole blood TNF production was evaluated. MAIN RESULTS: The capacity of trauma patients' whole blood to produce TNF was reduced for 2 to 6 days after trauma and was equally evident for both TNF release and TNFalpha mRNA levels. The reduction of TNF coincides with the appearance of an inhibitory activity for TNF production in trauma patients' sera. No correlation was found between the inhibitory activity and soluble TNF receptors, endotoxin-neutralizing molecules, inhibitory cytokines (interleukin 10 and transforming growth factor beta), or prostaglandins. CONCLUSIONS: Major trauma leads to the appearance of a circulating inhibitory activity for TNF synthesis that may potentially contribute to an anti-inflammatory response in patients with multiple trauma. The elucidation of its structural and functional properties may contribute to the understanding of the pathogenesis of severely injured patients. PMID- 9420100 TI - L-arginine decreases alveolar macrophage proinflammatory monokine production during acute lung injury by a nitric oxide synthase-dependent mechanism. AB - BACKGROUND: Recent clinical reports indicate that inhaled nitric oxide (NO) reduces lung parenchymal inflammation during acute lung injury; however, the mechanism of its protective effects remains incompletely understood. We hypothesized that the provision of substrate for local NO production (L-arginine) would reduce alveolar macrophage proinflammatory monokine production during endotoxin (ETX)-induced acute lung injury. Our purposes were to (1) determine alveolar macrophage tumor necrosis factor alpha (TNFalpha) and interleukin 1beta (IL-1beta) production after ETX-induced acute lung injury; (2) determine the effect of L-arginine on alveolar macrophage TNFalpha and IL-1beta production in ETX-induced acute lung injury; and (3) determine whether L-arginine's effects on the alveolar macrophage are mediated by NO. METHODS: Rats received ETX (0.5 mg/kg intraperitoneal (i.p.)) or vehicle, with or without (1) L-arginine supplementation (300 mg/kg i.p.) and (2) nitric oxide synthase inhibition (N(G) monomethyl-L-arginine, 30 mg/kg i.p.). Four hours later, alveolar macrophage were harvested by bronchoalveolar lavage and incubated at 10(6) cells/mL + 1 microg/mL phorbol myristase acetate for 24 hours. Cell-free supernatants were collected and assayed (enzyme-linked immunosorbent assay) for TNFalpha and IL-1beta. RESULTS: Sublethal ETX increased alveolar macrophage capacity to produce TNFalpha and IL 1beta (p < 0.05, analysis of variance and Bonferroni/Dunn). L-Arginine decreased alveolar macrophage TNFalpha and IL-1beta release during acute lung injury. Concurrent inhibition of nitric oxide synthase abrogated L-arginine's protective effects, suggesting that L-arginine's anti-inflammatory effects are mediated by NO. CONCLUSIONS: (1) L-Arginine is an immunomodulating nutritional supplement; (2) L-arginine decreases alveolar macrophage proinflammatory monokine production during ETX-induced acute lung injury by a nitric oxide synthase-dependent mechanism; and (3) the provision of exogenous substrate for local NO production may reduce inflammation during acute lung injury. PMID- 9420102 TI - Early prediction of prolonged ventilator dependence in thermally injured patients. AB - BACKGROUND: Recent studies suggest that when prolonged ventilator dependence (PVD) can be predicted in trauma or intensive care unit patients, early tracheostomy may reduce hospital stay and improve utilization of resources. This study was performed to develop criteria predictive of PVD (> 14 days) in burn patients. METHODS: We reviewed burn patients aged > or =16 years admitted between 1990 and 1994 who required ventilator support for > or =3 days. Using the variables full-thickness burn size, age, inhalation injury, and worst PaO2/FiO2 on ventilator day 3, an equation predicting PVD was created using logistic regression. The equation was tested by applying it to 1995 patients. RESULTS: When a probability of >0.5 was considered predictive of PVD, the equation correctly predicted PVD in 82% of 1990 to 1994 patients (n = 110) and 90% of 1995 patients (n = 29). CONCLUSION: PVD in burn patients can be predicted using objective variables in the early postburn period. Predictions can be used to select patients for prospective studies of early tracheostomy. PMID- 9420101 TI - The effect of blood transfusion on susceptibility to bacterial infection in genetically defined mouse models. AB - BACKGROUND: Blood transfusions suppress immune function and increase susceptibility to infection, but the effects are not consistent. STUDY DESIGN AND METHODS: Genetically defined mouse strains with the same or different haplotypes were used as blood transfusion recipients and donors. Transfused animals were subjected to cecal ligation and puncture (CLP) and followed for survival or were injected intravenously with Candida albicans to follow clearance of the Candida from the kidneys. RESULTS: BALB/c (H-2d) mice transfused with C3H/HeJ (H-2k) or DBA/2 (H-2d) blood followed by CLP showed significantly lower survival (7 and 10%) than mice transfused with syngeneic blood (61%) or saline controls (56%). Lower survival was also observed in C3H/HeJ (H-2k) mice transfused with BALB/c (H 2d) blood and subjected to CLP (25%) compared with syngeneic transfusion (80%) or saline controls (70%). C57BL/6J (H-2b) mice showed minimal increases in mortality after CLP after transfusion with blood from C3H/HeJ (H-2k) (60% survival), DBA/2 (H-2d) (70% survival), or BALB/c (H-2d) mice (90% survival). When C. albicans was infused intravenously into transfused mice, a similar pattern of altered resistance to infection was found. CONCLUSION: The ability of blood transfusions to increase susceptibility to bacterial infection appears to be dependent on genetic factors unrelated to the major haplotype. PMID- 9420103 TI - Inhaled nitric oxide in acute respiratory distress syndrome. AB - BACKGROUND: Inhaled nitric oxide has been shown to improve oxygenation in select patients with acute respiratory distress syndrome (ARDS). OBJECTIVE: The purpose of this study was to evaluate the clinical response to four concentrations of inhaled nitric oxide (NO) in 20 patients with ARDS. METHODS: All patients with ARDS were eligible for the study. ARDS was defined as (1) the presence of a predisposing factor; (2) a PaO2/FiO2 ratio < 200; (3) bilateral infiltrates on chest radiograph; and (4) absence of evidence of congestive heart failure and pulmonary artery wedge pressure < 18 mm Hg. Patients received each of four doses (1, 15, 30, and 60 ppm) in random order, each for a 3-hour period. Cardiovascular variables were continuously monitored, and arterial and mixed venous blood gas measurements were obtained at 30 minutes and 3 hours. RESULTS: Thirteen of the 20 patients demonstrated a significant increase in their PaO2/FiO2 (> 20% increase) when treated with inhaled NO. The administration of inhaled NO was associated with an increase in oxygenation at doses of 1, 15, and 30 ppm, but not 60 ppm. Increasing NO dose to more than 1 ppm did not significantly improve response. Mean pulmonary artery pressure decreased with increasing NO concentration, but this did not reach statistical significance. Nine of the 13 responding patients and 2 of the 7 nonresponding patients survived. CONCLUSION: Inhaled NO was successful in increasing PaO2/FiO2 by > 20% in 65% of the surgical patients in this trial. Response to NO could not be predicted by initial PaO2/FiO2 or pulmonary artery pressures. A trial of inhaled NO at a dose of < 10 ppm may be helpful in ARDS patients requiring increasing FiO2 and positive end-expiratory pressure. PMID- 9420104 TI - Inhibition of xanthine oxidase does not influence immunosuppression after hemorrhagic shock. AB - BACKGROUND: Resuscitated hemorrhagic shock causes global ischemia reperfusion with generation of toxic oxygen metabolites. We hypothesized that the immunosuppression that follows hemorrhagic shock may be linked to this process. METHODS: Forty-five male Sprague-Dawley rats (weight, 250-300 g) were bled to a mean arterial pressure of 30 mm Hg for 60 minutes, then were resuscitated with three times the maximum blood loss of lactated Ringer's solution. Immune response was assessed by splenocyte proliferation and interleukin-2 (IL-2) production 72 hours after hemorrhage. Allopurinol (50 mg/kg) was given after hemorrhage and immediately before resuscitation. RESULTS: Hemorrhagic shock caused significant decreases in splenocyte proliferation (cpm: (157,880 +/- 22,068 (mean +/- SD) vs. 37,787 +/- 15,849) and IL-2 production (1/2 max U/ml: 79.6 +/- 7.9 vs. 48.0 +/- 7.7) (both p < 0.05). Hepatic xanthine oxidase was significantly increased with hemorrhage and resuscitation. Hepatic xanthine oxidase activity after hemorrhage and resuscitation was significantly decreased after treatment with allopurinol (74.2 +/- 41.7 vs. 9.2 +/- 9.40). Allopurinol did not affect splenocyte proliferation (cpm: 21,875 +/- 9,316) or IL-2 production (1/2 max U/ml: 45.0 +/- 7.1). CONCLUSIONS: These results demonstrate that inhibition of xanthine oxidase by allopurinol after hemorrhagic shock did not affect splenocyte proliferation or IL-2 production. We conclude that the immunosuppression after hemorrhagic shock is not dependent on xanthine oxidase-induced production of toxic oxygen metabolites. PMID- 9420105 TI - The impact of the quantity of skeletal injury on mortality and pulmonary morbidity. AB - OBJECTIVE: To determine if the quantity of skeletal injuries (and the timing to fixation) increases the mortality or pulmonary morbidity in patients with and without chest injuries. DESIGN: Retrospective analysis of trauma registry. Statistical analysis with multiple logistic regression and chi(2) analysis. METHODS: Looking specifically at adult patients (> 16 years), skeletal injury was quantified by determining the presence or absence of a fracture in specific body regions (humeri, forearm, femur, tibia, spine, and pelvis) for a maximum of 10 skeletal injuries. The timing of fixation for fractures was categorized as < 24 hours, < 48 hours, < 72 hours, < 5 days, > 5 days, or no fixation. Chest injuries and pulmonary morbidity were based on the accepted list of complications reported in the literature. RESULTS: Three groups were analyzed according to the presence or absence of a chest or skeletal injury: those without skeletal injury (group NSI, n = 59), those without chest injuries (group NCI, n = 108), and those with both skeletal and chest injuries (group B, n = 59) Pulmonary Complications: When all patient groups (NCI, NSI, and B) were pooled, greater chest injury (p < 0.0008), greater skeletal injury (p < 0.02), and delayed fixation (p < 0.04) were associated with increased risk of developing a pulmonary complication. In the group of patients without a chest injury (NCI), this risk was associated with greater head injury (p < 0.005) and greater skeletal injury (p < 0.04), whereas in the group without a skeletal injury (NSI), only chest injury demonstrated significance (p < 0.05). When both skeletal and chest injuries were present, greater head injury (p < 0.03) and fixation time (p < 0.03) increased the risk of developing a pulmonary complication. Mortality: With all patients pooled (NCI, B, and NSI), head injury (p < 0.02), abdominal injury (p < 0.012), and fixation time (p < 0.01) were risk factors. In patients without a chest injury (NCI), none of the indexed variables were associated with mortality. In patients without a skeletal injury (NSI), greater head injury (p < 0.01), greater chest injury (p < 0.01), and greater abdominal injury (p < 0.04) were risk factors for mortality. When both chest and skeletal injuries were present (B), only head injury (p < 0.0003) was associated with mortality. The prevalence of mortality and pulmonary complications were compared between groups NCI, NSI, and B. Group NCI had fewer pulmonary complications (p < 0.004) than the other groups (difference not significant). When examining mortality, group NCI had less mortality than groups NSI and B. CONCLUSION: The combination of skeletal and chest injuries does not seem to amplify the pulmonary morbidity and mortality compared with chest injury alone. The quantity of the skeletal injury and the time to fixation of structures affecting mobilization seem to have an effect on pulmonary morbidity and mortality. Better scientific studies on the effects of skeletal injury and timing to fixation in relation to pulmonary morbidity and mortality are required. PMID- 9420106 TI - A modification of the injury severity score that both improves accuracy and simplifies scoring. AB - OBJECTIVES: The Injury Severity Score (ISS) has served as the standard summary measure of anatomic injury for more than 20 years. Nevertheless, the ISS has an idiosyncrasy that both impairs its predictive power and complicates its calculation. We present here a simple modification of the ISS called the New Injury Severity Score (NISS), which significantly outperforms the venerable but dated ISS as a predictor of mortality. DESIGN: Retrospective calculation of NISS and comparison of NISS with prospectively calculated ISS. MATERIALS AND METHODS: The NISS is defined as the sum of the squares of the Abbreviated Injury Scale scores of each of a patient's three most severe Abbreviated Injury Scale injuries regardless of the body region in which they occur. NISS values were calculated for every patient in two large independent data sets: 3,136 patients treated during a 4-year period at the American College of Surgeons' Level I trauma center in Albuquerque, New Mexico, and 3,449 patients treated during a 4-year period at the American College of Surgeons' Level I trauma center at the Emanuel Hospital in Portland, Oregon. The power of NISS to predict mortality was then compared with previously calculated ISS values for the same patients in each of the two data sets. MEASUREMENTS AND MAIN RESULTS: We find that NISS is not only simple to calculate but more predictive of survival as well (Albuquerque: receiver operating characteristic (ROC) ISS = 0.869, ROC NISS = 0.896, p < 0.001; Portland: ROC ISS = 0.896, ROC NISS = 0.907,p < 0.004). Moreover, NISS provides a better fit throughout its entire range of prediction (Hosmer Lemeshow statistic for Albuquerque ISS = 29.12, NISS = 8.88; Hosmer Lemeshow statistic for Portland ISS = 83.48, NISS = 19.86). CONCLUSION: NISS should replace ISS as the standard summary measure of human trauma. PMID- 9420107 TI - Analysis of heart-rate variability: a noninvasive predictor of death and poor outcome in patients with severe head injury. AB - BACKGROUND: Analysis of heart-rate variability (HRV) is a promising new technique for noninvasive quantification of autonomic function. We measured HRV in patients with severe head injury to assess its potential as a monitoring tool. METHODS: Analysis of HRV was prospectively done on all intensive care unit patients. Concurrent data on intracranial pressure (ICP) and cerebral perfusion pressure (CPP) were collected. Registry data were reviewed to identify patients with severe head injury, defined as Head/Neck Abbreviated Injury Scale score > or = 4. Mortality, likelihood of discharge to home, ICP, and CPP were compared between patients with abnormal HRV and those without. RESULTS: Low HRV was associated with increased mortality and decreased rate of discharge to home. Abnormal HRV was associated with episodes of increased ICP and decreased CPP. CONCLUSION: Assessment of HRV is a noninvasive method that can be widely used. Abnormal HRV was associated with poor outcome and altered cerebral perfusion. Monitoring of HRV may improve outcome by allowing earlier detection and treatment of intracranial pathology. PMID- 9420108 TI - Validation of an outcome prediction model for critically ill trauma patients without head injury. AB - BACKGROUND: The Acute Physiology and Chronic Health Evaluation (APACHE) II system is inaccurate in predicting the risk of death in trauma patients, especially those without head injury. Using multivariate analysis of the APACHE II system in a development set, a new predictive equation was modeled. The four variables that were independently associated with mortality were PaO2/FiO2 ratio, mean arterial pressure, temperature, and the need for inotropic support. This model was tested prospectively in an independent validation set of 300 patients. METHODS: Risk of death was calculated using the APACHE II system with the diagnostic category of multiple trauma and weighting for operative intervention as required. The new model was similarly assessed using the four predictor variables and their beta coefficients for each mechanism of injury and the entire group. The predicted risk of death derived by both models was compared with the observed mortality rate. Discrimination was calculated using a 2 x 2 decision matrix with a decision threshold of r = 0.5 and receiver operating characteristic curves. Calibration was assessed graphically and by statistical correlation. RESULTS: The observed mortality rate was 28.3% and the predicted mortality risk was 27.4% for the model and 6.26% for APACHE II. The sensitivity and specificity of the model were 58.8 and 90.7%, and the sensitivity and specificity of APACHE II were 1.2 and 100%. The areas under the receiver operating characteristic curves were 0.84 and 0.78 for the model and the APACHE II system, respectively. Calibration of the model was superior within all deciles of risk (model, R2 = 0.93, p < 0.001; APACHE II, R2 = 0.82, p = 0.02). CONCLUSION: The model accurately predicted the risk of death for the entire group. It is superior to the APACHE II system and is the highest reported sensitivity for 24-hour intensive care unit predictive models that have been applied to the critically injured. PMID- 9420109 TI - A comparison of the association of helicopter and ground ambulance transport with the outcome of injury in trauma patients transported from the scene. AB - INTRODUCTION: Comprehensive emergency medical services and helicopter aeromedical transport systems have been developed based on the principle that early definitive care improves outcome. The purpose of this study was to compare outcomes between patients transported by helicopter and those transported by ground. METHODS: Data were obtained from the North Carolina Trauma Registry for the period between 1987 and 1993 on all patients transported by helicopter and ground admitted to one of the eight state designated trauma centers. Study patients included only those who were transported directly from the scene of injury to the trauma center (interhospital transfers were excluded). Mortality (outcome) was compared after patient stratification by injury severity and transport time, using Cochran-Mantel-Haenszel statistics and logistic regression derived probabilities of survival. RESULTS: One thousand three hundred forty-six patients (7.3% of the total) were transported from scene to trauma center by helicopter and 17,144 were transported by ground. In patients transported by helicopter, the mean Trauma Score was lower (12 +/- 3.6) versus 14.3 +/- 3.6 (p < 0.001) and the mean Injury Severity Score was higher (17 +/- 11.1) versus 10.8 +/ 8.4 (p < 0.001). A trend toward increased survival was observed among patients transported by helicopter with a higher Injury Severity Score. Statistical significance was achieved only for patients with a Trauma Score between 5 and 12 and Injury Severity Score between 21 and 30. CONCLUSION: The large majority of trauma patients transported by both helicopter and ground ambulance have low injury severity measures. Outcomes were not uniformly better among patients transported by helicopter. Only a very small subset of patients transported by helicopter appear to have any chance of improved survival based on their helicopter transport. This study suggests that further effort should be expended to try to better identify patients who may benefit from this expensive and risky mode of transport. PMID- 9420110 TI - Closed humeral shaft fractures: a prospective evaluation of surgical treatment. AB - OBJECTIVE: We tried to define the roles of the rigid dynamic compression plate (DCP) and the semi-rigid Ender nail (EN) in the treatment of closed humeral shaft fractures. DESIGN: A prospective, randomized clinical study was performed with detailed comparison parameters. MATERIALS AND METHODS: Ninety-one closed humeral shaft fractures were treated. Randomly, 30 humeri were treated with open reduction and internal fixation with DCP and no bone grafting (BG), 29 were treated with the same procedure but with BG, and 32 were treated with closed reduction and internal fixation with Ender nails. The average follow-up period was 32 months (range, 13-54 months). MEASUREMENTS AND MAIN RESULTS: In the group with DCP without BG, the average blood loss was 270 mL, operation time was 92 minutes, hospital length of stay was 6.5 days, and union time was 12.5 weeks. In the group with DCP with BG, the average blood loss was 325 mL, operation time was 108 minutes, hospital length of stay was 6.9 days, and union time was 9.4 weeks. In the EN group, the average blood loss was 114 mL, operation time was 54 minutes, hospital length of stay was 5.6 days, and union time was 9.9 weeks. Analysis of variance and Fisher's exact test were used to evaluate the statistical significance. CONCLUSION: In our experience, for humeral shaft fractures fixed surgically, EN is better than DCP without BG. When DCP is chosen for the means of fixation, prophylactic BG is recommended, especially in cases with more comminution. PMID- 9420111 TI - Adequacy and efficacy of lateral cervical spine radiography in alert, high-risk blunt trauma patient. AB - OBJECTIVE: The purpose of this study was to determine the adequacy and accuracy of lateral cervical spine radiographs in the initial evaluation of alert, high risk trauma patients evaluated at a Level I trauma center. METHODS: Data were obtained retrospectively through review of trauma service admissions from January 1, 1994, to July 31, 1995. Included were all patients triaged to a trauma response team with age > 15 years, Glasgow Coma Scale score > 13, and blunt mechanism of injury. Lateral cervical spine radiograms were obtained routinely before secondary survey and were reviewed for technical adequacy (all seven cervical vertebrae, C7/T1 interspace). The presence of cervical symptoms (pain, tenderness, neurologic deficits) was recorded. Sensitivity and specificity were calculated for lateral cervical spine radiography and cervical symptoms in predicting the presence of cervical spine injury. Bayesian analysis, which allows for the current probability of occurrence to be factored by previously reported probabilities of occurrence, was used to determine the negative predictive probability of lateral cervical spine radiography and absence of cervical symptoms to predict the absence of injury to the cervical spine. RESULTS: Three hundred fifty-three patients received lateral cervical spine radiograms, of which 223 (63%) were determined to be adequate for interpretation. Cervical symptoms were present in 77 patients (20%). Only 32 (42%) of this group's lateral cervical spine radiograms were adequate. Nine patients (2.4%) had acutely fractured cervical vertebrae or ligamentous disruption. Lateral cervical spine radiography showed the injury in only six of these patients. The sensitivity, specificity, and negative predictive probability for lateral cervical spine radiography were 67, 58, and 1.4%, respectively, and for absence of cervical symptoms, 89, 81, and 0.32%, respectively. CONCLUSION: The higher accuracy and lower negative predictive probability make the absence of cervical symptoms in the alert, high risk, blunt trauma patient a better screening test than lateral cervical spine radiography. We suggest that lateral cervical spine radiography is not needed in the initial evaluation of alert patients who have sustained blunt trauma. PMID- 9420112 TI - Resection of the radial head: an alternative to use of a prosthesis? AB - Prosthetic substitution of the head of the radius after comminuted fractures has not yielded satisfactory long-term results. For this reason, resection of the radial head is to be preferred in many cases. In the period from 1971 through 1985, 23 resection of the radial head were performed. After a follow-up interval of 17 years, on average, we did a personal reexamination of all patients. Using the Morrey score, we obtained 18 very good, 4 good, and 5 satisfactory results, but not a single poor outcome. On the basis of reports in the literature and our own results, we recommend resection of the radial head, especially in cases in which stable osteosynthesis cannot be achieved, as long as there are no better long-term results of prosthetic substitution of the radial head. PMID- 9420113 TI - The accuracy of the CAGE, the Brief Michigan Alcoholism Screening Test, and the Alcohol Use Disorders Identification Test in screening trauma center patients for alcoholism. AB - OBJECTIVE: To evaluate the accuracy of questionnaire screening instruments to identify lifetime alcohol dependence among trauma center patients. METHODS: The study was conducted at a Level I trauma center between September 1994 and November 1996. Patients meeting eligibility requirements (> or = 18 years old, admission from injury scene, > or = 2 days of hospitalization, intact cognition) were evaluated for alcohol abuse and dependence. Screening instruments consisted of the CAGE, the Brief Michigan Alcoholism Screening Test, and the Alcohol Use Disorders Identification Test. Screening results were compared with lifetime alcohol dependence diagnoses made using the in-depth Psychoactive Substance Use Disorders section of the Structured Clinical Interview. Accuracy was quantified as sensitivity, specificity, positive/negative predictive values, and receiver operating characteristic curves (used to calculate area under the curve). RESULTS: Of the 1,118 patients studied, lifetime alcohol dependence was diagnosed by Structured Clinical Interview in 397 (35.5%), and abuse was diagnosed in 90 (8.1%) others. The CAGE was the best predictor of lifetime alcohol dependence, i.e., had the largest area under the curve (93%) and the highest sensitivity (84%), specificity (90%), positive predictive value (82%), and negative predictive value (91%). Among patients testing positive for alcohol, 63% had a lifetime alcohol dependence diagnosis. CONCLUSION: The CAGE is an efficient screening test to detect alcohol dependence in trauma center populations. It should be used in combination with alcohol testing to identify patients at risk of alcohol use problems. PMID- 9420114 TI - Forgoing medical treatment in severe facial trauma. PMID- 9420115 TI - Ventral tension pneumothorax. PMID- 9420116 TI - Damage control in a trauma patient with ureteric injury. PMID- 9420117 TI - Splenosacral sinus after splenorrhaphy: case report. PMID- 9420118 TI - Torsion of a wandering spleen after blunt abdominal trauma. PMID- 9420120 TI - Providing high-quality patient care in the face of cost containment and shortening length of stay requirements. PMID- 9420119 TI - A quicker saphenous vein cutdown and a better way to teach it. AB - The saphenous vein cutdown has long been a mainstay for venous access in the trauma patient. During the past several years, however, its popularity and frequency of use have declined markedly. Percutaneous femoral catheterization using the Seldinger approach has essentially replaced the cutdown as the method of choice for gaining vascular access in most circumstances. There remains a group of critically ill patients, however, often without palpable femoral pulses, in whom percutaneous femoral lines are difficult if not impossible to place. The saphenous vein cutdown can be lifesaving in these patients, provided that physicians can preserve the skill to place the cutdowns efficiently. This will be even less likely in time because the new revision of the Advanced Trauma Life Support textbook will make the saphenous vein cutdown an "optional" skill to be taught at the discretion of the course director. Presented is a faster, more efficient saphenous vein cutdown procedure using a Seldinger wire-guided dilator. Also presented is an effective, inexpensive model for teaching this procedure and for skill preservation. PMID- 9420122 TI - Albumin leak in urine early predicts pulmonary dysfunction and ARDS. PMID- 9420121 TI - Cost-effective prevention of pulmonary embolus in high-risk trauma patients. PMID- 9420123 TI - Development of allergic disease in children. PMID- 9420124 TI - A deficient capacity to produce interferon-gamma: is it a risk for asthma and allergies? PMID- 9420125 TI - T-helper polarization in atopic disease--how early does it occur? PMID- 9420126 TI - Peanut allergy--current status and future challenges. PMID- 9420127 TI - The development of allergy in high-risk children. AB - BACKGROUND: It is uncertain as to what extent the development of allergic disease in childhood is predictable during early infancy. A number of environmental factors have been suspected of increasing the risk of acquiring allergy, but the evidence is conflicting. OBJECTIVE: To observe the development of atopy and allergic disease in a cohort of high-risk children so as to determine the importance of certain environmental factors and to study the relationship between early and later manifestations. METHODS: A cohort of infants, all at high risk of allergy, was followed up from birth to the age of 7 years. In half, selected at random, cow's milk protein was avoided for 4 months. Skin-prick tests were performed and serum IgE measured in infancy and at 7 years, when an AlaTOP test was also performed. RESULTS: Skin sensitivity to egg in the first year of life was strongly associated with eczema, asthma, mite sensitivity and serum IgE at the age of 7 years, when mother's atopic history was associated with AlaTOP status, father's atopic history with skin sensitivity, and male sex with both. Maternal smoking during pregnancy was associated positively with IgE at 3 months and negatively with skin sensitivity at 7 years. The development of allergy was unrelated to infant feeding method or number of older siblings. CONCLUSION: Allergic disease in childhood is to a large degree determined before birth or during infancy. PMID- 9420129 TI - TH2-polarized immunological memory to inhalant allergens in atopics is established during infancy and early childhood. AB - BACKGROUND: There is increasing evidence that the T-cell reactivity to environmental allergens underlying expression of allergic disease in adulthood, develops initially during childhood. However, there is little information available on the kinetics of these early responses, or on the patterns of cytokine production during this period. OBJECTIVE: The purpose of this study was twofold: to obtain further information on the reported differences between responses to food versus inhalant allergens during early childhood, and to ascertain the age-range over which T-cell responses to inhalant allergens become polarized towards the TH2 cytokine profile, in potentially atopic children. METHODS: In vitro cytokine responses to house dust mite (HDM) and egg (OVA) were assessed by semiquantitative RT-PCR in panels of 2- and 5-year-old children and adults; lymphoproliferative responses to OVA were subjected to epitope analysis. RESULTS: At age 2 years IL-4/IL-5 responses to HDM grouped with positive atopic family history, and by age 5 years cytokine responses correlated strongly with individual SPT reactivity to HDM. In contrast, OVA responses were restricted to weak and transient IL-5 signals in the 2-year-old family history positive group. Lymphoproliferation assays performed in parallel indicate a log-scale greater postnatal expansion of T-cell reactivity to the inhalant allergen; preliminary epitope analysis of OVA responses indicate that the number of OVA epitopes recognised decrease during early childhood. CONCLUSIONS: Inhalant allergen specific in vitro cytokine production associated with positive skin-prick test (SPT) reactions, one of the hallmarks of adult atopy, manifests in children at or before 5 years of age; additionally, cytokine responses in SPT negative 5 year olds are restricted to IFNgamma, as per normal adults. In contrast, T-cell responses to a typical food allergen appear to be deleted during early childhood. PMID- 9420128 TI - Interleukin-4, interferon-gamma and interleukin-5 in peripheral blood of children with moderate atopic asthma. AB - BACKGROUND: In asthmatic inflammation, TH2 cells play an important role. TH2 cells specifically secrete cytokines like IL-4 and IL-5. IL-4 stimulates IgE production and IL-5 is involved in hemopoiesis, chemotaxis, priming and activation of eosinophils. IFNgamma, produced by TH1 cells, has an inhibitory action on IgE production. OBJECTIVES: To investigate the TH1/TH2-cell pattern in the cytokine production of peripheral blood of asthmatic children. We determined IL-4, IFNgamma and IL-5 in serum and in supernatants of unstimulated and stimulated (24 h with Concanavaline A) cultures of peripheral blood mononuclear cells (PBMCs) in 22 children with moderate asthma (mean age 9.3 years) and in 17 healthy controls (mean age 10.3 years). All children visited the out-patient department (OPD) where history taking, physical examination and blood sampling took place. Children younger than 8 years of age performed symptom and peak flow registration during 1 week after the visit to the OPD. RESULTS: The number of eosinophils were significantly higher in children with asthma, compared with healthy controls. The concentration of IFNgamma in supernatants of cultures of stimulated PBMCs was significantly lower and the ratio of IL-4/IFNgamma was significantly higher in children with asthma compared with healthy controls. The FEV1 was directly and IgE was inversely related to the concentration of IFNgamma in supernatants of cultures of stimulated PBMCs. CONCLUSION: IFNgamma may play an important role in the pathophysiology of childhood atopic asthma. PMID- 9420130 TI - Asthma among secondary schoolchildren in relation to the school environment. AB - BACKGROUND: Poor indoor air quality has been suggested to be related to the increase in the prevalence of asthma that has occurred in the western world, especially among children and young persons. Apart from the home, school is the most important indoor environment for children. OBJECTIVES: The aims were to study the prevalence of current asthma among secondary pupils and its relationship to the school environment, but also to personal factors and domestic exposures. METHODS: Data on asthmatic symptoms, other health aspects, and domestic exposures were gathered using a questionnaire which was sent to 762 pupils in the seventh form (13-14 years old) in 11 randomly chosen schools in the county of Uppsala in Sweden. Pupils answering 'yes' to having had asthma diagnosed by a physician, and having had recent asthma attacks, or who used asthma medication were defined as having current asthma. Data on exposures at school were gathered by measurements in 28 classrooms. The relationship between asthma and exposures was analysed by multiple logistic regression. RESULTS: The questionnaire was completed by 627 (82%). Current asthma was found among 40 pupils (6.4%). Current asthma was more common in those who had an atopic disposition, or food allergy, or who had attended a day care centre for several years. Controlling for these factors, current asthma was related to several factors in the school environment. There were more pupils with current asthma in schools that were larger, had more open shelves, lower room temperature, higher relative air humidity, higher concentrations of formaldehyde or other volatile organic compounds, viable moulds or bacteria or more cat allergen in the settled dust. CONCLUSIONS: Although the pupils attended school for a minor part of their time, our study indicates that the quality of the school environment is of importance and may affect asthmatic symptoms. PMID- 9420131 TI - Immunotherapy vs inhaled budesonide in bronchial asthma: an open, parallel, comparative trial. AB - BACKGROUND: Budesonide, an inhaled corticosteroid and specific immunotherapy, are both routinely used in the treatment of bronchial asthma. However, there are as yet, no studies comparing the effects of budesonide vs immunotherapy. OBJECTIVE: The aim of this study is to compare the effects of budesonide with immunotherapy in patients having perennial asthma. METHODS: This study is an open, parallel, comparative trial, in which 51 young patients were administered either immunotherapy or budesonide for 1 year and their global symptom scores and FEV1 values assessed. Both treatments were abruptly discontinued after 12 months and the effects of cessation analysed. RESULTS: The use of budesonide resulted in a faster and more striking improvement during the first few months as compared to immunotherapy, with an even more rapid decline in benefits on cessation of budesonide. Immunotherapy on the other hand, resulted in slow but steady improvement which did not decline as rapidly as budesonide on cessation. CONCLUSION: Although this was an open trial, it could be concluded that relief with inhaled corticosteroids in bronchial asthma is more rapid than immunotherapy; however the decline in benefit on cessation of inhaled corticosteroid is even more rapid, a phenomenon not seen with immunotherapy. PMID- 9420132 TI - Specific humoral immune responses in 12 cases of food sensitization to sesame seed. AB - BACKGROUND: Hypersensitivity to sesame seeds is becoming increasingly frequent, probably owing to the larger use of this compound in international food. OBJECTIVES: This study investigated serum responses of 12 sesame sensitized patients (seven with food allergy, five with food sensitization), to a sesame protein extract, and attempted at identifying sesame major antigens. METHODS: Sesame protein extracts were prepared from black, white and brown sesame seeds. Electrophoretic analysis showed similar protein patterns in the three extracts, and proper preservation of the proteins integrity. The brown sesame extract was used to set-up an ELISA assay and measure serum levels of antisesame IgG, IgA, IgM and IgE in 12 samples from sesame-sensitized individuals and six controls. It also allowed to perform western blot analyses in order to investigate the molecular weight of sesame proteins recognized by IgG, IgA and IgE. RESULTS: Nineteen protein bands were observed upon polyacrylamide gel electrophoresis of the sesame protein extracts. Using this whole extract in ELISA, significant antisesame IgG, IgA and IgE-responses were observed in the serum of sensitized individuals, different from the lower signals obtained with control samples. Western blot analysis demonstrated highly polymorphic IgG and IgA responses and a more restricted IgE response pattern, suggesting that two proteins, respectively, 14 kDa and 25 kDa are mostly involved in sesame IgE-dependent hypersensitivity, the 25 kDa band presenting several characteristics of a major allergen. CONCLUSIONS: This study reports novel information on the possible involvement of a 25 kDa sesame protein in IgE-dependent hypersensitivity to sesame seeds. PMID- 9420133 TI - Expression of interleukin (IL)-4 and IL-5 proteins in asthma induced by toluene diisocyanate (TDI). AB - BACKGROUND: TDI-induced asthma exhibits clinical, functional and morphological similarities with allergen-induced asthma, suggesting that an immunological mechanism is involved in the sensitization to TDI. In vitro studies using the technique of cloning lymphocytes demonstrated that a great proportion of T-cell clones derived from bronchial mucosa of subjects with TDI-induced asthma produced IL-5 and interferon-gamma, but not IL-4, upon in vitro stimulation. OBJECTIVES: To investigate in vivo the role of IL-4 and IL-5 on the inflammatory response of the bronchial mucosa to TDI in sensitized subjects, we performed a quantitative analysis of bronchial biopsies. METHODS: We obtained bronchial biopsies from six subjects with TDI asthma 48 h after an asthmatic reaction induced by TDI challenge (challenged group), in six subjects with TDI asthma 1-4 weeks after the last exposure to TDI (chronic group), and in six non-asthmatic controls. The number of eosinophils, mast cells, T-lymphocytes, and IL-4 and IL-5 protein positive cells was determined by immunohistochemistry in the area 100 microm beneath the epithelial basement membrane. RESULTS: The characteristic increase of submucosal eosinophils, but not of mast cells and T-lymphocytes, was observed in the subjects with TDI-induced asthma when compared with controls. No differences were detected between the two groups of asthmatics. In the subjects with TDI induced asthma, cell immunoreactivity for IL-5 was increased when compared with normal controls. There was no difference in the expression of IL-5 protein between challenged and chronic asthmatics. In contrast, the expression of IL-4 protein was increased only in the asthmatic subjects tested after recent exposure to TDI. CONCLUSIONS: We demonstrated that TDI asthma 48 h after specific bronchial challenge was associated with increased numbers of cells expressing IL 4 and IL-5, whereas chronic TDI asthma was associated with increased expression of IL-5, but not of IL-4. The results suggest that subjects who developed TDI asthma exhibit increased production of IL-5 even in the absence of a recent trigger by the exogenous sensitizer and that production of TH2-like cytokines in TDI-induced asthma may not always be co-ordinately regulated in vivo. PMID- 9420134 TI - Characterization of allergens in Apiaceae spices: anise, fennel, coriander and cumin. AB - BACKGROUND: Symptoms elicited by IgE-mediated food allergy range from mild local to severe systemic reactions. Allergens in spices are particularly dangerous due to their hidden presence in many dishes. OBJECTIVES AND METHODS: According to clinical observations, mugwort and birch pollen allergy, and hypersensitivity to spices are frequently associated, but the crossreacting compounds were not defined so far. We tested sera of 15 patients who experienced adverse reactions to spiced food and characterized their IgE-binding patterns on anise, fennel, coriander and cumin extracts through immunoblot and inhibition experiments. RESULTS: The use of anti-Bet v 1 (MoAb) and anti-profilin (rabbit) antibodies revealed the presence of crossreacting allergens in the tested spice extracts. Inhibition experiments showed that IgE-binding to allergens in Apiaceae spices could be blocked by preincubation of sera with rBet v 1 or rBet v 2 (birch profilin). Moreover, we detected crossreacting allergenic molecules in the molecular weight range of 60 kDa. IgE-binding to spice allergens occurred only with sera of 10/15 (66%) patients with allergy to pollen (birch, mugwort) and/or celeriac. In five out of 15 (33%) patients with a history of adverse reaction to spices, but without pollen and celeriac allergy, no IgE-binding to any spice protein could be demonstrated. It is possible that these clinical reactions could be elicited by other types of hypersensitivity (Type II, III, IV), however, as spices contain highly reactive substances, the symptoms may most likely be classified as food-intolerant. CONCLUSIONS: Bet v 1- and profilin-related allergens may, besides higher molecular weight allergenic molecules, be responsible for Type I allergy to anise, fennel, coriander or cumin, members of the Apiaceae. PMID- 9420135 TI - Cloning and high level expression of Cynodon dactylon (Bermuda grass) pollen profilin (Cyn d 12) in Escherichia coli: purification and characterization of the allergen. AB - BACKGROUND: Profilin, an actin-binding protein, was previously described as a panallergen which is involved in about 20% of the crossreactivity found among pollen and food allergic patients. This allergen is usually under-represented in natural extracts used for allergy diagnosis. OBJECTIVES: To obtain an immunologically active and soluble recombinant profilin from Cynodon dactylon pollen which could be used for diagnostic and therapy. METHODS: Isolation of cDNA clones was performed by polymerase chain reaction amplification using degenerate primers. Expression in Escherichia coli BL21 (DE3) was carried out using vector pKN172, and the expressed product was isolated by affinity chromatography on poly L-proline-Sepharose. RESULTS: Four cDNA inserts coding for Cynodon dactylon (Bermuda grass) pollen profilin (Cyn d 12) were cloned and sequenced. Full-length C. dactylon profilin gene was expressed in Escherichia coli as non fusion protein. Induced cells could produce high amounts of recombinant Cyn d 12, and after a single purification step on poly (L-proline)-Sepharose, up to 45 mg of pure allergen per litre culture could be obtained. The reactivity of recombinant Cyn d 12 with IgE antibodies present in sera from Bermuda grass-allergic patients is comparable to that of the natural Bermuda grass allergen. Recombinant Bermuda grass pollen profilin was shown to share B-epitopes with sunflower profilin. CONCLUSIONS: Our results showed that this heterologous expression system and purification procedure are suitable for the production of large amounts of pure allergen which can be used for the characterization of allergenic epitopes recognized by T and B cells and finally for diagnostic and therapeutic purposes. PMID- 9420136 TI - Comparison of a radioallergosorbent (RAST) inhibition method and a monoclonal enzyme linked immunosorbent assay (ELISA) for aeroallergen measurement. AB - BACKGROUND: Mouse and rat urinary proteins are potent occupational allergens for exposed personnel. Methods of measuring airborne allergens differ greatly, and reported levels of allergens vary considerably between laboratories. OBJECTIVES: To compare the values obtained using two different methods of allergen detection. METHODS: Air samples were collected in rat rooms in Sweden and the United Kingdom at 2 L/min on to polytetrafluoroethylene (PTFE) filters and extracted in buffer containing 0.5% v/v Tween 20. Airborne rat urinary allergen (RUA) was measured in all samples by both RAST inhibition using a polyclonal human serum pool (UK) and a two monoclonal antibody sandwich ELISA employing antibodies specific for Rat n 1.02 (alpha2u-globulin) (Sweden). RESULTS: The two methods gave values which were correlated (r2 log values = 0.72, P<0.0001), but differed by several orders of magnitude (median [range] ratio of RAST inhibition/ELISA = 316 [7-26(80)]. There was a systematic bias: as the absolute values increased, the difference in the measurements increased. The rat urine standards used were antigenically similar. CONCLUSIONS: A large contrast in RUA values obtained from the two assays was observed in this study. This may be primarily due to methodological differences, but variations in antibody specificities or composition of allergenic epitopes in the air samples may contribute. The results demonstrate that standardization of methods and antibodies is necessary before interlaboratory comparisons can be made. PMID- 9420137 TI - Substance P is generated in vivo following nasal challenge of allergic individuals with bradykinin. AB - BACKGROUND: Bradykinin, a potent inflammatory mediator, is released during allergic and non-allergic rhinitis and asthma in man. Nasal bradykinin challenge induces a dose-dependent plasma leakage into the nasal cavity and relevant symptoms of rhinitis. OBJECTIVE: We now report on substance P generation during nasal bradykinin challenge in vivo. METHODS: The effect of locally applied bradykinin on substance P generation was studied in nine individuals, allergic to grass pollen and six non-allergic controls. In the allergics TAME-esterase activity, histamine and substance P concentrations were measured in nasal lavages and correlated to the clinical symptoms. RESULTS: Substance P concentrations in nasal lavages increased in a dose-dependent fashion during nasal bradykinin challenge in both groups. In the allergic group Substance P-increases correlated with the production of TAME-esterase activity (r = 0.9, P < 0.05) whereas these allergic individuals did not produce any histamine increases. The generation of substance P and the increase of TAME-esterase activity was associated with the onset of clinical symptoms. Correlation of oedema and hypersecretion to substance P were significant by linear regression analysis (r = 0.88, P < 0.005 and r = 0.89, P < 0.02, respectively). Bradykinin induced irritations like burning and itching were short-term and rare. Serial dilutions of nasal washes produced Substance P-RIA displacement curves that paralleled the standard curve and recovery of standard substance P that was added to nasal washes was 76 +/- 4% (mean +/- SEM), n = 8. CONCLUSION: Bradykinin induces in vivo a dose-dependent plasma leakage into the nasal cavity without affecting mast cells, but stimulates nerve endings resulting in the release of the neuropeptide substance P. PMID- 9420138 TI - Immunolocalization of cytokines to mast cells in normal and allergic conjunctiva. AB - BACKGROUND: Recently, the potential role of mast cells in allergic reactions has been extended by the discovery that these cells synthesize, store and secrete multifunctional cytokines. Seasonal allergic conjunctivitis is characterized as an immediate hypersensitivity reaction, in which allergen binds to specific IgE on mast cells, leading to release of pre-formed and newly synthesized inflammatory mediators. OBJECTIVE: In this study we aimed to localize the cytokines IL-4, IL-5, IL-6, IL-8 and TNF alpha to conjunctival mast cells and to examine the relationship between mast cell-associated cytokines and allergic conjunctivitis. METHODS: Immunohistochemistry was performed on serial sections of conjunctival biopsies from patients with seasonal allergic conjunctivitis, in and out of the hay fever season, as well as from non-allergic volunteers. RESULTS: IL 4, IL-5, IL-6 and TNF alpha were localized to mast cells in normal and allergic conjunctiva. IL-8 was localized to mast cells in two patients with seasonal allergic conjunctivitis, one during and the other outside the pollen season. Using the monoclonal antibody 3H4, which identifies the secreted form of IL-4, biopsies from patients with active seasonal allergic conjunctivitis contained a significantly higher proportion of mast cells positive for IL-4, than those from out-of-season patients (P=<0.016). There was no difference between the two groups in the number of mast cells immunostained by the antibody 4D9 which identifies the stored form of IL-4. CONCLUSIONS: These results suggest that conjunctival mast cells can store a range of multifunctional cytokines and release IL-4 during active disease, which may give them an important role in upregulating allergic inflammation in the conjunctiva. PMID- 9420139 TI - A new role for interleukin-7 in the induction of LFA-1 and VLA-4 adhesion molecules in Phorbol 12myristate 13acetate activated CD4+ CD23+ T-cell subset. AB - BACKGROUND: The low affinity receptor for IgE, CD23, has been described in several pathological conditions. However, the factors involved in the upregulation or downregulation of this receptor are still debated. METHODS AND RESULTS: We studied the effect of interleukin 7 (IL-7) on the expression of CD23 in normal PBT cells stimulated with PMA + Ca2. The data indicate that activated PB-T cultured in the presence of IL-7 showed an increased expression of CD23. The induction of IL-7 on CD23 production appears to be independent of IL-2, IL-4, IL 9, IL-15. Indeed, the addition of specific MoAbs anti-IL-2, IL-4, IL-9, IL-15 or anti-IL2R was unable to block the effect of IL-7 on CD23. The addition of IL-7 to a specific subset CD4+ CD23+ was able to augment the adhesiveness of T cells to parenchymal cell monalayers. The use of different cytokine (IL-2, IL-4, IL-9, IL 15) resulted in no increase of adhesiveness. In contrast the addition of IL-7 to a different T-cell subset (i.e. CD4+ CD23-) was unable to rescue the lack of adhesiveness observed in these cells. Blocking experiments with MHM6 MoAb were able to drastically reduce the adhesiveness observed in CD4+ CD23+ subsets. The presence of LFA-1 and VLA-4 adhesion molecules were responsible for the augmented adhesiveness of activated CD4+ CD23+ T cells cultured in the presence of IL-7. Blocking experiments with anti-LFA-1, VLA-4, anti-LFA-1beta plus VLA-4alpha MoAbs or anti-ICAM-1 MoAb added to the monolayers resulted in a complete inhibition of adhesion to parenchymal monolayers. In contrast, the addition of anti-IL-7 or anti-IL-7R MoAbs were able to block the augmented adhesiveness of CD4+ CD23+ cells to monolayers observed in the presence of IL-7. CONCLUSION: Taken together these findings point to the likelihood that IL-7 is responsible for the observed quantitative difference in the level of adhesion molecules and may open a new role of CD23 in the immune regulation. PMID- 9420140 TI - Association between inflammation and epithelial damage-restitution processes in allergic airways in vivo. AB - BACKGROUND: Associations between allergen challenge-induced sites of epithelial damage and the distribution of leucocytes and extravasated plasma remain unexplored. OBJECTIVE: To study neutrophils, eosinophils, and fibrinogen at allergen challenge-induced patchy epithelial damage-restitution sites in guinea pig trachea. METHODS: After local challenge tracheal tissue (cryo sections and whole-mounts) and lumen (selective tracheal lavage) were examined at 1, 5, and 24 h. Eosinophils, neutrophils and fibrinogen were identified by histochemistry. RESULTS: Neutrophils increased markedly in tracheal lavage fluids and in tissue and were strongly associated with the challenge-induced epithelial craters of damage-restitution. At 1 and 24 h eosinophils were increased in the tracheal lumen whereas the surrounding tissue displayed a reversed pattern. Gels rich in fibrinogen, neutrophils, and eosinophils were present in epithelial crater areas, protruding into the lumen. Clusters of free eosinophil granules, Cfegs, released through lysis of eosinophils, and neutrophils with long cytoplasmatic protrusions abounded in these crater areas. CONCLUSION: The present findings provide important new insights into allergic airways where sites of epithelial damage restitution processes emerge as the major loci for eosinophil, neutrophil, and plasma protein activities, the latter likely causing leukocyte adhesion and activation in vivo. The distribution of eosinophils in this study suggests roles of these cells both in airway mucosa and in regional lymph nodes. Based on the present study we also propose that lysis of eosinophils and Cfegs generation are a major paradigm for activation of these cells in vivo. PMID- 9420141 TI - Notice of duplicate publication. PMID- 9420142 TI - Enhancement of resolution of low molecular weight RNA profiles by staircase electrophoresis. AB - Stable low molecular weight (LMW) RNA comprises molecules used in the taxonomy of microorganisms and in studies on the microbial diversity of populations. However, the use of electrophoretic techniques has been hampered due to the low resolution obtained with techniques used for the separation of this kind of molecule. In this work we develop an electrophoretic method (staircase electrophoresis) that increases the resolution of the technique. This improvement in the resolution adequately resolves the three zones that integrate the profiles of LMW RNA: ribosomic 5S RNA (5S rRNA), class 2 transfer RNA (tRNA), and class 1 transfer RNA, allowing the technique to be applied to taxonomic studies (diagnostic, the identification of the individuals), phylogenetic studies and studies on naturally occurring microbial populations. PMID- 9420143 TI - Reorientation of large DNA molecules in concentrated polyacrylamide solution during crossed-field electrophoresis. AB - Recently, we found that, in concentrated neutral solutions, DNA molecules migrate in linear conformation under steady electric field. In this paper, we report the conformational change of DNA during 120 degree crossed-field electrophoresis in the same polymer solution. We found that, in concentrated polyacrylamide solutions, the reorientation process of DNAs becomes simple: the DNA goes back along the previous track and the reorientation time is longer for larger DNA. Such a backtrack motion has been thought to be an essential motion for the separation of DNA fragments in pulsed field gel electrophoresis. We expect that this phenomenon is useful for a more efficient separation technique of large DNAs than the current pulsed field gel electrophoresis. PMID- 9420144 TI - A biostatistical study into the efficiency of individualism using nonisotopic chemiluminescent-enhanced NICE multilocus DNA probes. AB - The efficiency of individualisation using nonisotopic chemiluminescent- enhanced probes (NICE) was investigated by analysing DNA fingerprints obtained from 190 unrelated Caucasians. Novel analysis of the scoring procedure enabled us to include the comparison of 585 pairs of samples for each of two probes. When the results of NICE probes 33.6 and 33.15 were combined, the mean percentage band share between two unrelated individuals was 16.8% and the mean number of bands identified in an individual DNA fingerprint was 54.8. Results were compared with those obtained using isotopically labelled probes and suggest that the two labelling systems gave similar efficiencies for differentiating between individuals. Analysis of DNA fingerprints from 37 family trios (mother, child and father groups) gave a mutation rate of 0.10% when using NICE probes. The two labelling systems compared were equally efficient in establishing family relationships. PMID- 9420145 TI - D12S67, a bipartite locus: differential amplification of parts of the nucleotide sequence reveals considerable polymorphism. AB - The STR system D12S67 was amplified by polymerase chain reaction (PCR) and analyzed by denaturing gel electrophoresis followed by silver staining. Among 133 DNA samples from Japanese individuals, 11 alleles were observed and the heterozygosity was 80%. When sequences of the alleles were compared, each allelic segment contained 35-45 gata or gaca tetranucleotide repeats. Although a (gaca)n repeat block was concentrated in a defined region, nine different blocks of (gata)n repeats were observed separated by the (gaca)n repeat and single copy sequences. In addition, the allelic differences result from the total number of repeats of at least three (gata)n and the (gaca)n repeat regions. Novel primers overlapping part of the sequence with each other were constructed in the center of the D12S67 sequence, and the 5' and 3' segments were amplified by PCR. Both of these segments were highly polymorphic and showed heterozygosities of 77 and 78% with 7 and 10 alleles, respectively. The genotypes of the two polymorphisms were not concordant with the original polymorphism and the combination of the 5' and 3' side segment polymorphisms enabled further detailed classification of the D12S67 locus by simple comparison of the PCR product sizes. The number of the allele types increased up to 35 and the heterozygosity to 93%. PMID- 9420146 TI - Detection and quantitative characterization of artificial extra peaks following polymerase chain reaction amplification of 14 short tandem repeat systems used in forensic investigations. AB - Detection on automated DNA sequencers of polymerase chain reaction (PCR) products of tetra- and penta-nucleotide short tandem repeat (STR) loci frequently reveals one or more extra peaks along with the true, major allele peak. The most frequent extra peak pattern is a single smaller peak which is one repeat unit shorter than the true allele peak. The existence of such artificial peaks is of special importance when the methods are used for forensic investigations because the artificial extra peaks may simulate true alleles when samples containing mixtures of DNA from different individuals are analyzed. We have investigated the relative levels of formation of extra peaks in 14 STR marker systems. We found that not only the parameters of the PCR but also factors determining the stringency during the post-PCR and pre-electrophoresis handling of samples were of importance for the formation of extra peaks. In our hands, the amounts of extra peaks were reduced (i) if the samples were effectively denatured immediately before loading, (ii) if they contained substantial amounts of formamide (i.e. > or = 50%), and (iii) if the temperature of the electrophoresis gel was above a certain level (i.e. > or = 43 degrees C). The results suggest that extra peaks may in part be due to re-annealing of the PCR product under suboptimal conditions. When efforts had been made to reduce the post-PCR formation of extra peaks, the relative peak areas of the extra peaks ranged from 1% to 17% of those of the true alleles. Similar results were obtained when the PCR products were analyzed under native conditions. Low-copy genome analysis excluded that somatic heterogeneity of the STR regions caused the extra peaks. The systems HumVWA31A, HumFibra/FGA, and D21S11 were especially affected by low-stringency conditions, while Hum-TH01, HumCD4, and D12S391 were virtually unaffected by low-stringency conditions. Replacement of the Taq DNA polymerase with DNA polymerases with lower processivity resulted in higher levels of extra peaks. Our results support the hypothesis that extra peaks are produced due to slipped-strand mispairing. PMID- 9420147 TI - The fifth allele of the human deoxyribonuclease I (DNase I) polymorphism. AB - The fifth allele, DNASE1*5, of human deoxyribonuclease I (DNase I) has been discovered. Polymerase chain reaction fragments containing exon 5 of the DNase I gene were screened for DNA polymorphism using single-strand conformation polymorphism (SSCP) analysis. DNAs from 114 unrelated Japanese and 81 German individuals were tested and a new variant was detected. By DNA sequencing analysis, this variant was found to be caused by a heterozygous G-A transition at nucleotide position 1227 that results in a Val to Met substitution at amino acid position 92 of the mature enzyme. The nucleotide substitution was also confirmed by mismatched polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) analysis. Genotyping of the variant could be carried out by three independent reactions based on PCR amplification, and phenotyping by isoelectric focusing followed by immunostaining. The results supported the presence of the fifth codominant allele, DNASE1*5, which generates a new isozyme. PMID- 9420148 TI - Simple and rapid determination of the acetaldehyde dehydrogenase (ALDH2) genotypes by nonradioactive single-strand conformation polymorphism analysis. AB - The genotyping of mitochondrial acetaldehyde dehydrogenase (ALDH2) is very important in alcohol studies. We describe an ALDH2 genotyping method based on nonradioactive single-strand conformation polymorphism (SSCP) analysis on mini gels following amplification with a mutated primer set. The three ALDH2 genotypes were clearly and unambiguously distinguished. This method was applied to the ALDH2 genotyping of 129 unrelated Japanese. The allele frequency of ALDH2*2 was calculated to be 0.271, which was consistent with the previous data. The method proved to be simple, rapid and reliable, and dispensed with isotopic reagent and expensive restriction enzymes and equipment. The SSCP method described here is valuable in routine ALDH2 genotyping. PMID- 9420149 TI - Further genetic heterogeneity in acatalasemia. AB - A T-deletion at position 10 of exon 4 for catalase gene was reported as a novel mutation, causing a new genetic type of acatalasemia in Japan. This mutation, destroying a Hinf1 recognition site, was searched for in Hungarian acatalasemic (2) and hypocatalasemic (22) patients and in controls (27) by Hinf1 digestion and sequence analyses of a 203 bp polymerase chain reaction (PCR) product containing the entire exon 4. The Hinf1 polymorphism did not reveal any difference between controls and hypocatalasemic as well as acatalasemic patients. These results were confirmed by sequence analyses showing the T nucleotide for the two acatalasemic and for one unrelated hypocatalasemic patient, as well as for two controls. These findings represent further evidence that acatalasemia is heterogeneous at the DNA level. PMID- 9420150 TI - Ampholyte dissociation theory and properties of ampholyte aqueous solutions. AB - Two different approaches (stepwise and parallel mechanism) for describing the dissociation of amphoteric protolytes are described. Additionally, the classification of the possible model on the basis of the ampholyte lifetime state is proposed. The possibility of application of different schemes is discussed. It is suggested that the correct description may be based only on highly relaxing models. The formulas describing the properties of ampholyte solutions are derived for the case of two ionogenic groups. It is demonstrated that the incorrect choice of the dissociation model may result in essential mistakes in calculation of such values as buffer capacity, electrophoretic mobility, and conductivity. On the basis of the theory here developed, the interpretation of natural pH-gradient conductivity is given. Some terminology questions are also discussed. PMID- 9420151 TI - Improved sensitivity of detection with the commercial automated gel electrophoresis (HPGE-1000) apparatus through modification of its optical system. AB - In a representative application to a fluorescently detectable protein of commercial automated gel electrophoresis apparatus (HPGE-1000, LabIntelligence, Belmont, CA) the sensitivity of detection by fluorescence was significantly increased by elimination of the mirror below the gel tray. That increase in detection sensitivity is due to a decrease of fluorescent background noise by nearly one order of magnitude, overcompensating a decrease in signal by a factor of two. The resulting increase in signal/noise ratio, i.e., detection sensitivity, should allow for lowered sample loads by which the band width is reduced with benefits to resolution. PMID- 9420152 TI - The effect of sodium tetradecyl sulfate on mobility and antigen detectability of microtubule proteins in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. AB - Several factors been reported to influence the mobility of polypeptide in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) including the brand of SDS. Using microtubule proteins from axonemes of Lytechinus pictus and Spisula solidissima sperm and meiotic spindles of Spisula solidissima we demonstrate that the change in mobility was caused by sodium tetradecyl sulfate (STS), a major contaminant of many commercial SDS brands. We also examined the use of sodium tetradecyl sulfate and different SDS brands as a tool in extracting more information from immunoblot studies. Commercial SDS containing contaminants other than sodium tetradecyl sulfate reduced or eliminated the immunosignal from certain polypeptides and the loss of antigenicity could not even be recovered by immunoblot under "renaturing" conditions. It can thus be concluded that STS can be useful in separating and identifying comigrating polypeptides and in detecting additional immunobands in immunoblots. PMID- 9420153 TI - Nonenzymatic chemiluminescent detection and quantitation of total protein on Western and slot blots allowing subsequent immunodetection and sequencing. AB - We have studied the light emission efficiency of proteins labeled with different fluorescent dyes chemically excited by the bis(2,4,6-trichlorophenyl)oxalate (TCPO)-H2O2 reaction. Using this peroxyoxalate chemiluminescence system, the best results were obtained with proteins covalently labeled with 2-methoxy-2,4 diphenyl-3(2H)-furanone (MDPF). Blotted proteins on polyvinylidene difluoride (PVDF) membranes can be labeled rapidly with MDPF. Our results demonstrate that energy from the excited intermediate produced in the TCPO-H2O2 reaction can be efficiently transferred to MDPF-labeled proteins in solution and on PVDF membranes. Although this nonenzymatic chemiluminescent system produces a background emission that reduces the sensitivity, the method developed in this work allows detection of 5 ng of protein in blots after 5 min exposure to X-ray film. Chemiluminescence of MDPF-labeled proteins on Western and slot blots may also be detected and quantified using a charge-coupled device (CCD) camera or a storage phosphor imaging system. This chemiluminescent method allows the staining of the total electrophoretic pattern but does not preclude further N-terminal sequencing and immunodetection of specific bands. PMID- 9420154 TI - A sensitive electrophoretic method for the quantification of myosin heavy chain isoforms in horse skeletal muscle: histochemical and immunocytochemical verifications. AB - In adult horses, three myosin heavy chain (MyHC) isoforms can be identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunohistochemistry using specific anti-MyHC monoclonal antibodies. This report studies the suitability of a consistent SDS-PAGE technique for quantifying MyHC profiles in homogenized cryostate sections of equine gluteus medius muscle biopsies (n = 18). The method used (previously described by R. J. Talmadge and R. R. Roy; J. Appl. Physiol. 1993, 75, 2337-2340) resolved MyHCs in three bands: I, IIB or IIX, and IIA from the fastest to the slowest migration band. The success rate of the protocol for yielding three well-differentiated MyHC bands was 100% and a subsequent quantification by densitometry for each MyHC isoform was obtained in all 18 muscle biopsies. The results obtained with this electrophoretic method were compared with routine myofibrillar adenosine triphosphatase histochemistry and immunohistochemistry using specific anti-MyHC monoclonal antibodies. The percent composition of the three electrophoretically separated MyHC isoforms (I, IIA and IIB or IIX) showed strong positive correlation with percentages of the area occupied in the biopsies by the three major fiber types (I, IIA, and IIB) identified histochemically (r = 0.96, P < 0.001) and immunohistochemically (r = 0.94, P < 0.01). It can be concluded that the electrophoretic method used here for measuring MyHC content is a valid alternative for muscle fiber typing in horses. As it is less costly and time consuming than both qualitative histochemistry and immunohistochemistry, the method offers new prospects for application in equine experimental studies and veterinary medicine. PMID- 9420155 TI - Gel electrophoresis of DNA fragments in narrow-bore capillaries. AB - In this work, we studied the behavior of electrophoretic columns, having an inner diameter (ID) of 2-10 microm, filled with a cross-linked polyacrylamide gel matrix. The usefulness of these columns for DNA sequencing is discussed. Evaluation of column performance included tests of gel stability and migration time reproducibility. Confocal laser-induced fluorescence (LIF)-detection was employed utilizing a 488 nm argon ion laser for separations of C- and T terminated DNA Sanger fragments. Reducing the inner diameter of the column from 50 microm to 10 microm resulted in an approximately eightfold increase in lifetime, under conditions in which the columns were subjected to a field strength of 1000 V/cm. The 10 microm ID columns were utilized for separation of Sanger fragments, and adequate detection sensitivity was obtained by stacking of the fragments from a deionized sample solution. A linear algorithm for retention data synchronization between individual electropherograms was employed to provide a route towards a reliable automated base calling protocol. PMID- 9420157 TI - Micropreparative capillary electrophoresis of DNA by direct transfer onto a membrane. AB - We have developed a new technique for the collection of DNA fragments separated by capillary electrophoresis, by direct transfer from the capillary outlet to a positively charged membrane. Transfer and post-run detection of two different nonradioactively labeled DNA standards, ranging in size from 150 bp to 2 kbp and 120 bp to 23 kbp are presented, and discussed. Capillary electrophoresis with direct blotting presents several advantages over the blotting from gels: the separation is faster and requires less manual steps, the resolution is higher, and each DNA fragment is collected into a very concentrated spot on the membrane due to the small surface of the capillary outlet and to a design of the collection device inducing a refocusing of field lines across the hybridization membrane. Therefore, very small amounts of DNA (in the pg range) can be detected. This fraction collection makes further analysis of the sample possible, e.g. by hybridization, thus suppressing one of the major present limitations of the capillary electrophoresis technique for DNA analysis. PMID- 9420156 TI - The characterization of composite agarose/hydroxyethylcellulose matrices for the separation of DNA fragments using capillary electrophoresis. AB - Mixtures of the polysaccharide derivatives, 19% hydroxyethylated SeaPrep agarose (SP-AG) and hydroxyethylcellulose (HEC), in aqueous buffer solutions are applied for the first time to the separation of DNA fragments using capillary electrophoresis (CE). These matrices form unique size-sieving networks that allow the separation of a wide size range of DNA fragments in a single analysis. Relative to their homogeneous counterparts, the composite separation matrices provide enhanced selectivity properties of DNA fragments, especially for fragments greater than 1000 base pairs (bp) in length. Additionally, the effects on separation performance of capillary temperature, the incorporation of a DNA intercalator, and applied field strength are demonstrated. Solution viscosity measurements of the homogeneous and composite matrix solutions were made in order to establish the entanglement threshold concentrations for the unique size sieving solutions. The relatively low solution viscosities of the composite separation matrices allow reproducible replacement of the separation matrix between analyses. The mechanism of separation of DNA fragments for the composite matrices is proposed. PMID- 9420158 TI - Capillary electrophoresis of DNA restriction fragments: effect of polymer properties. AB - The mechanism of DNA separation by dilute polymer solutions in capillary electrophoresis is not well understood. To provide information on the effect of polymer properties on DNA separations, four polymers that differ in size, shape and stiffness were examined. Hydroxyethylcellulose of high molecular weight provides excellent separation of large DNA fragments (2027 bp - 23,130 bp). Polyvinylpyrrolidone separates DNA from 72 bp to 23 kbp; star-poly(ethylene oxide) and linear poly(ethylene oxide) provide separation of fragments to 1353 bp. PMID- 9420159 TI - System peaks in capillary zone electrophoresis. 3. Practical rules for predicting the existence of system peaks in capillary zone electrophoresis of anions using indirect spectrophotometric detection. AB - A theoretical and experimental study of the existence and evolution of system peaks in capillary zone electrophoresis (CZE) with indirect spectrophotometric detection is presented with respect to the effect of the number of coions present in the background electrolyte (BGE). It is shown that in BGEs having only one coion (i.e., the UV-absorbing probe anion), the sample produces only negative peaks due to each analyte anion and no system peaks, with the number of sample peaks corresponding to the number of analytes present in the sample injected. In BGEs containing two coions, a sample with one analyte anion produces one negative indirect detection peak and one system peak. The transition between BGEs having one coion and those with two coions has also been studied and it has been shown that an addition of ca. 5% of the second coion to a single coion BGE causes the resulting BGE to behave macroscopically as a regular two-coion BGE. A descriptive model is proposed, based on transient isotachophoresis (transient stacking) of the sample species and of the coion from the BGE which has the closest mobility to the sample ion. This model explains qualitatively the formation and evolution of the sample peak (containing the sample species and being detected by indirect detection due to displacement of the UV-absorbing probe in its zone) and the system peak (containing no sample species and being a vacancy in the continuum of coions of the BGE). It is shown that the system peak may be positive or negative as it corresponds to the situation where the vacancy of one component of the BGE results in an enhanced concentration of the other component. It has been demonstrated that the system peak is created by a vacancy of that component of the BGE which has the greatest difference in mobility relative to that of the sample species. On indirect detection in BGEs containing two coions the sample displaces predominantly the BGE coion which has a mobility closest to that of the analyte anion. In systems with BGEs containing two coions, a sample having n analytes produces n sample peaks and one system peak, the sign and magnitude of which are dependent on the sum of the UV absorbances of the analytes involved. The effect of bicarbonate from atmospheric CO2 has also been studied and it has been shown that weakly alkaline BGEs with a single anionic UV-absorbing coion, such as those currently used for anionic analyses with indirect detection, may suffer from the presence of system peaks due to bicarbonate. PMID- 9420160 TI - Some properties of a measure of resolution in gel electrophoresis and capillary zone electrophoresis. AB - The most commonly used measure of resolution for chromatographic and electrophoretic separations does not take into account the possibility of there being different amounts of each of the molecular species. A modification of a measure of resolution recently suggested by Aldroubi and Garner (BioTechniques 1992, 13, 620-624) can incorporate this effect explicitly. Their criterion for resolution is based on the time to observe a valley of specified magnitude separating two peaks. We examine how this measure depends on different physically relevant parameters that characterize the system. PMID- 9420161 TI - Generation of peptide maps by capillary zone electrophoresis in isoelectric iminodiacetic acid. AB - Capillary zone electrophoresis in stationary, isoelectric buffers is a novel method for generating peptide maps of protein digests. The buffer system developed is composed of iminodiacetic acid (IDA), whose physico-chemical parameters were found -- by theoretically modeling and experimental verification - to be: pI 2.23 (at 100 mM concentration), pK1 = 1.73 and pK2 = 2.73 (no attempts were made at measuring the pK of the primary amino group, since such a low pI value would be compatible with any pK value of the basic group, down to as low as pK 5.5). IDA is compatible with most hydro-organic solvents, including trifluoroethanol (TFE), up to at least 40% v/v, typically used for modulating peptide mobility. In naked capillaries, a buffer comprising 50 mM IDA, 10% TFE and 0.5% hydroxyethylcellulose (HEC) allows generation of peptide maps with high resolution, reduced transit times and no interaction of even large peptides with the wall. However, the best background electrolyte was found to be a solution of 50 mM IDA in 0.5% HEC and 6-8 M urea, one of the best solubilizers of proteins and peptides known. In this last electrolyte system, peptide maps of beta-casein digests (known to contain also very large peptides, up to 6000 Da) could be generated with excellent resolution and half the transit times as compared with the standard buffer adopted in peptide analysis (80 mM phosphate buffer, pH 2.0). IDA thus appears to be another valid isoelectric buffer system, operating in a different pH window (pH 2.33 in 50 mM IDA) as compared to the other amphotere previously adopted (50 mM Asp, pH 2.77) for the same kind of analysis.